PMID- 7470346 TI - Needle in the heart. AB - A patient accidentally drove a darning needle through her chest wall into her left ventricle. After referral five weeks later as possible case of infective endocarditis, the needle was detected and removed successfully by open heart surgery. It was localised precisely by echocardiography. PMID- 7470345 TI - Right atrial outlet atresia with straddling left atrioventricular valve. A form of double outlet atrium. AB - We present three hearts showing right atrial outlet atresia with straddling left atrioventricular valve; clinical data of two of the patients are reported. For semantic and practical reasons we believe the terms tricuspid or mitral atresia should not be used in these cases, and atrial outlet atresia is preferable; for similar reasons we prefer to use stradding left or right atrioventricular valve. These anomalies represent an interesting form of double outlet atrium and diagnosis clinically may be possible if the existence of the anomaly is considered. Differentiation from common atrioventricular canal with atrial outlet atresia is important since total repair, which is possible for this last anomaly, does not seem feasible in cases like ours. Classifications of tricuspid atresia may have to be enlarged to include cases with double outlet from either ventricular chamber or anatomically corrected malposition. Characterisation of the right ventricular chamber is difficult; we believe that description of its morphology and the type and mode of its connections should be used. PMID- 7470347 TI - Double Wenckebach phenomenon in atrioventricular node and His bundle. Electrophysiological demonstration in a case of atrial flutter. AB - In a case of atrial flutter with a 9:2 atrioventricular response, the only possible way to explain the conduction pattern was 3:1 block in the atrioventicular node (which is 3:2 Wenckebach sequence in the N zone and a 2:1 block at the junction of the node with the bundle of His) plus 3:2 Wenckebach sequence distal to the H deflection. The recording of the His bundle deflection confirmed this analysis. PMID- 7470348 TI - Metabolism and anesthesia. PMID- 7470349 TI - The glucose-fatty acid cycle. Relationship between glucose utilization in muscle, fatty acid oxidation in muscle and lipolysis in adipose tissue. PMID- 7470350 TI - The glucose-fatty acid-ketone body cycle. Role of ketone bodies as respiratory substrates and metabolic signals. PMID- 7470351 TI - The regulation of glucose production. The role of liver glycogen and gluconeogenesis in the liver and kidney cortex. PMID- 7470352 TI - Antagonism of ketamine by 4-aminopyridine and physostigmine. PMID- 7470353 TI - Lumbar extradural analgesia. PMID- 7470354 TI - Tympanic membrane rupture during nitrous oxide anaesthesia. PMID- 7470355 TI - Plasma cholinesterase and malignant hyperthermia. PMID- 7470356 TI - Protection of the brain from ischaemia. PMID- 7470357 TI - Effect of some i.v. anaesthetic agents on canine gastrointestinal motility. AB - Thiopentone 20 mg kg-1, ketamine 8 mg kg-1 and minaxolone 2 mg kg-1 were administered to fasting greyhound dogs. Mechanical and electrical activities from stomach, duodenum, jejunum and ileum were recorded using strain gauge force transducers and implanted bipolar electrodes. Thiopentone and minaxolone caused intense activity in the duodenum and jejunum (phase I and phase II of the interdigestive cycle), but not the stomach or ileum. The activity following injection of thiopentone or minaxolone was prevented by premedication with either atropine 0.05 mg kg-1 or pentolinium 0.2 mg kg-1. Ketamine had no influence on gastrointestinal activity or the response to thiopentone or minaxolone. None of these drugs altered the basal electrical rhythm of the intestine. PMID- 7470358 TI - Evaluation of the efficacy of an active scavenger for controlling air contamination in an operating theatre. AB - Contamination of the air with halothane in a poorly ventilated ENT theatre was found to be very great (median concentration 449 micrograms litre-1, 177 samples), but was reduced significantly (P less than 0.005) by the use of an active scavenger of patients' expired gases (median concentration 159 micrograms litre-1, 168 samples). For about 18% of the time when the theatre was in use, the halothane concentration in the air was greater than 1000 micrograms litre-1, whether or not the patients' expired gases were being scavenged. Although not measured quantitatively, this appeared to correspond approximately to the period of time in theatre when patients were not connected to the anaesthetic circuit. PMID- 7470359 TI - Malignant hyperthermia and the fluoride-resistant gene. AB - One hundred and six individuals from 33 families with a history of malignant hyperthermia have been investigated for plasma cholinesterase variants. An increased frequency of the fluoride-resistant gene has been found. Although an adequate explanation for our results is elusive, some hypotheses are discussed. PMID- 7470360 TI - Collaborative study of the frequency of the fluoride-resistant cholinesterase variant in patients with malignant hyperpyrexia. AB - Cholinesterase phenotyping studies have been carried out in two laboratories on blood specimens from 28 patients shown to be susceptible to malignant hyperpyrexia. In contrast to previously published reports, no evidence could be found for an increase in the frequency of the E1u E1f genotype above that known to occur in the general population. PMID- 7470361 TI - Rate of formation of norpethidine from pethidine. AB - The concentrations of norpethidine in blood were determined in patients recovering from abdominal surgery following both multiple i.m. injections (i.m. group) and the i.v. infusion (i.v. group) of pethidine. The rate of administration of pethidine averaged 25 mg h-1 in both groups. Concentrations of norpethidine were generally small compared with those of pethidine although the ratio of norpethidine to pethidine concentrations increased steadily over the 2 days studied. The rate of increase of blood concentration of norpethidine was 0.013 micrograms ml-1 h-1 in both groups of patients. No significant toxic effects were observed. PMID- 7470362 TI - Influence off haemodilution on the potency of neuromuscular blocking drugs. AB - The potencies of suxamethonium, pancuronium, and tubocurarine were increased, as shown by a parallel shift to the left of cumulative dose-response curves, following normovolaemic haemodilution before operation. This is attributed to an increase in cardiac output, to a decrease in protein binding and, in the case of suxamethonium, to a decrease in cholinesterase activity. PMID- 7470363 TI - Hexobarbitone disposition at different stages of intensive care treatment. AB - The pharmacokinetics of hexobarbitone were investigated in 22 patients in an intensive care unit. The results were compared with those obtained in a healthy group for three time periods: 3rd or 4th day (I), 5-8th days (II) and 13-29th days (III) of treatment. Hexobarbitone 7.32 mg per kg of body weight was administered by i.v. infusion in 60 min. At the end of the infusion, the mean plasma concentration of group I was 71% greater than control group; groups II and III were near to control. In all patients the post-infusion concentration-time course of hexobarbitone could be described by two-compartment kinetics. The biphasic decrease in plasma concentration of hexobarbitone was more rapid in groups II and III, than in either the control group or in group I. At the beginning of treatment patients generally showed an unchanged hexobarbitone half life, a slightly decreased plasma clearance, a reduction of approximately 50% of the initial distribution volume. (V1) and a reduction of 44% of the distribution volume at steady state. In groups II and III the mean of these values were comparable to control. However, plasma clearance had increased by about 87% in group II compared with control, and after 2-3 weeks (group III) by about 143%. Correspondingly, half-life was reduced by 45% and 58% respectively. The pharmacokinetics were not related to change in liver function. The presence of clinical and bacteriological signs of septicaemia was closely associated with an enhanced hexobarbitone clearance. PMID- 7470364 TI - Hypotension during and after operation in glucocorticoid-treated patients. AB - The frequency and likely aetiology of systemic arterial hypotension occurring either during or following operation, in patients receiving steroids was analysed retrospectively. Hypotension occurred during surgery in 29 of 250 operations. Although patients receiving steroids until shortly before operation appeared more likely to develop hypotension during operation the differences were eliminated when allowance was made for the severity of the disease. In almost half of the instances of hypotension during surgery there was no adequate explanation. Hypotension occurred after surgery on eight occasions; there was one case of acute adrenocortical insufficiency, six of various types of circulatory failure and in one the cause as uncertain. PMID- 7470365 TI - Plasma cholinesterase and malignant hyperthermia. PMID- 7470366 TI - Induction dose of midazolam. PMID- 7470367 TI - An infrequent cause of pulmonary atelectasis during mechanical ventilation. PMID- 7470368 TI - Sensitivity of the newborn to tubocurarine. PMID- 7470369 TI - Anaesthesia for laryngoscopy. PMID- 7470370 TI - Propranolol disposition in renal failure. AB - 1 Previous studies of propranolol disposition in renal failure have been conflicting. 2 Using simultaneous administration of [3H]-propranolol intravenously and unlabelled propranolol orally the principal determinants of drug distribution were calculated in normals, patients with severe renal impairment (creatinine clearance 14.5 +/- 2.8 ml/min) but not on haemodialysis and patients on haemodialysis (creatinine clearance less than 5 ml/min). 3 The effect of haemodialysis on propranolol binding and free fraction was also examined. The percentage of propranolol unbound rose from 7.1% to 9.9%. (P less than 0.001) 20 min following heparinization and beginning haemodialysis. This was accompanied by a large rise in free fatty acids from 0.567 +/- 0.059 to 3.326 +/- 0.691 mumol/ml (P less than 0.005). 4 The blood to plasma concentration ratios of propranolol were significantly higher in patients with renal failure (P less than 0.02) and on haemodialysis (P less than 0.001) and were significantly negatively correlated (P less than 0.001) with the haematocrit. 5 Although the half-life propranolol was significantly shortened in the patients with renal failure (P less than 0.02), there was no change in the apparent liver blood flow, extraction ratio or the principal determinants of steady-state drug concentrations in blood namely oral and intravenous clearance from blood. 6 There is, therefore, no pharmacokinetic basis to adjust the dosage of propranolol in patients with renal failure. PMID- 7470371 TI - Biphasic dose-related responses of the CNV (contingent negative variation) to I.V. nicotine in man. PMID- 7470372 TI - 'MacDope': a simulation of drug disposition in the human body: applications in clinical pharmacokinetics. AB - 1 We have described a novel approach to absorption, distribution, metabolism and elimination of drugs in which the patient is described using 23 Patient Factors and drugs by up to 50 Drug Factors. Kinetic behavior of a drug results from the interaction of patient and drug factors according to equations describing an eight compartment model. In this model non-linear processes (protein binding, hepatic drug metabolism and renal tubular transport) are handled by derivations of the law of mass action which have been generalised to permit the consequences of multiple drugs interacting at single macromolecular sites to be correctly calculated. 2 The mathematical description of this model is provided in a companion paper and solution of the equations is only possible using a digital computer. The computer programme is provided with an interactional format which makes operation independent of mathematical skills. Patients are defined by age, sex, height and weight with, or without, organ dysfunction; the programme then generates appropriate factors. Drug enters the system when a prescription is type on the keyboard and required Drug Factors are then retrieved from the disc flies. Drug concentrations in plasma or body fluids are given as simple graphs as a function of time, or in tabular form. 3 Any of the Patient or Drug Factors may be altered before, or during a run and up to three drugs may be simulated at one time thus permitting certain kinetic interactions to be examined. The scope of the simulator is illustrated using aspirin: pH dependent gastric absorption, first-order conversion of aspirin to salicylate, partly first order and partly saturable hepatic metabolism of salicylate, and the complex renal handling of this drug are all represented. Interaction of phenytoin with salicylate has been examined quantitatively to suggest limited clinical relevance for the observed displacement of phenytoin from serum albumin. The use of the simulator in a short course of pharmacokinetics is briefly described. PMID- 7470373 TI - Metabolism of an oral tryptophan load III: effect of a pyridoxine supplement. PMID- 7470374 TI - Actions of dopamine upon human urinary bladder muscle in vitro. PMID- 7470375 TI - Haemodynamic effects of diltiazem in healthy volunteers. PMID- 7470376 TI - Some effects of atropine on short-term memory. PMID- 7470377 TI - Comparative functional histopathology of human breast carcinoma xenografts. AB - A series of xenografts of human breast carcinomas has been established and serially transplanted in immune-suppressed mice. Certain structural and functional features of the original human tumours, including carcinoembryonic antigen and epithelial membrane antigen, continue to be expressed by the resulting xenografts. Stromal responses such as elastosis and oestrogen-receptor activity were lost by the xenografts. No metastases were detected in tumour bearing mice. This study suggests that xenografts may have some value in experimental pathology as one type of model of human breast carcinoma. PMID- 7470378 TI - Clonogenic cell survival in cryopreserved human tumour cells. AB - Cells from 3 human tumours have been grown in soft agar contained in Millipore diffusion chambers and implanted i.p. in mice. Clonal growth was obtained from fresh biopsy samples, from cryopreserved tissue, and from xenografts of the tissues in immune-suppressed mice. The radiosensitivities of a melanoma and an ovarian carcinoma were evaluated by in vitro irradiation before assay for colony formation. Xenografting did not modify the radiosensitivity of the melanoma. Cells from another tumour were exposed to Adriamycin or cyclophosphamide whilst contained within i.p. diffusion chambers; the sensitivity was similar for cryopreserved and xenografted cells. The results encourage further attempts to quantify the sensitivity of human tumour cells by these methods. PMID- 7470380 TI - A case-control study of head and neck cancer in the Republic of Ireland. AB - A retrospective case-control study of 200 patients with head and neck cancer, and 200 controls matched for age and sex, confirmed the importance of tobacco and alcohol consumption in the aetiology of malignant tumours of the upper gastrointestinal and upper respiratory tracts. A male-female ratio of 3:1 was found, and the association of smoking with laryngeal cancer and of alcohol with cancer of the tongue was particularly strong. A significant excess of alcohol related occupations was found among the cases. These findings are discussed. PMID- 7470379 TI - Malignant lymphoma and exposure to chemicals, especially organic solvents, chlorophenols and phenoxy acids: a case-control study. AB - A number of men with malignant lymphoma of the histiocytic type and previous exposure to phenoxy acids or chlorophenols were observed and reported in 1979. A matched case-control study has therefore been performed with cases of malignant lymphoma (Hodgkin's disease and non-Hodgkin lymphoma). This study included 169 cases and 338 controls. The results indicate that exposure to phenoxy acids, chlorophenols, and organic solvents may be a causative factor in malignant lymphoma. Combined exposure of these chemicals seemed to increase the risk. Exposure to various other agents was not obviously different in cases and in controls. PMID- 7470381 TI - Cancer mortality in Shanghai during the period 1963-77. AB - In Shanghai a population-based Cancer Registry has been in operation since 1963 covering the urban area, with total population of about 5.6 million. In this report methods of collecting cancer deaths and population data are described in detail, and cancer mortality data for the period 1963-77 presented. The main fatal cancers were those of stomach, lung, liver, oesophagus and colon-rectum; among females, in addition there were cancers of uterus and breast. During the 15 year period a rapid increase in cancer-mortality rate was seen for lung, colon rectum, pancreas in both sexes, and for bladder in males. A notable decrease in mortality rate for cancer of the uterus (mainly for cancer of cervix uteri) occurred. PMID- 7470382 TI - Successful immunotherapy with micrococcus, BCG or related polysaccharides on L1210 leukaemia after BCNU chemotherapy. AB - The experiments aimed at evaluating the optimal parameters in the chemo immunotherapeutic treatment of the L1210 lymphoid leukaemia grafted to [female BALB/c (H2d) X male DBA/2 (H2d)]F1 hybrid mice, hereafter referred to as CDF1 mice. In vitro irradiation of leukaemic ascites cells by X- or gamma-rays and subsequent inoculation in mice showed that optimum immunogenicity is radiation dose-dependent. Grafting mice with 10(7) leukaemic ascites cells irradiated at optimum dose (80 GyX- or gamma-rays) delays mortality of the animals when challenged later with untreated L1210 cells, but is unable to cure mice. By contrast, specific immunoprophylaxis induced by Micrococcus, complement triggering polysaccharides or BCG and irradiated leukaemic cells was able to protect mice against grafts of 10(4) L1210 cells. The i.p. route was notably superior to the i.v. route. When mice bearing advanced L1210 tumour were treated by chemotherapy (12 mg/kg of BCNU) on Day 6.5 after grafting 10(4) L1210 cells and subsequently treated by immunotherapy, a very high percentage (up to 90%) of mice with 10(8) leukaemic cells could be cured by repeated 1mg injections of bacterium or polysaccharide, and challenge with irradiated leukaemic cells was unnecessary. Because of the high cure rate obtained, the very regular response pattern and the non-pathogenicity, the bacterium Micrococcus lysodeikticus would seem a promising new candidate for chemo-immunotherapeutic antitumour strategies. PMID- 7470383 TI - Significant variation in mouse-skin aryl hydrocarbon hydroxylase inducibility as a function of the hair growth cycle. AB - An easy, rapid and improved technique for homogenizing whole skin is described. This technique consists of reducing skin to a powder in liquid N2 by using a metallic mortar, and homogenizing the powder in a Potter-Elvehjem tube. Using this homogenizing method, we have shown that skin AHH activity in C57BL/6K and C3H/Ico mice can be induced by i.p. injected or topically applied methylcholanthrene during a defined period of the hair growth cycle, i.e. between the 8th and 14th days after depilation (Stage 6 of the anagen period). In each experimental model, there is an optimal methylcholanthrene concentration which yields a maximum induction. Topical methylcholanthrene is also responsible for a smaller aryl hydrocarbon hydroxylase (AHH) induction when the chemical is applied the same day that the club hairs are plucked. On the other hand, skin AHH activity is never induced by methylcholanthrene in DBA/2J mice, a genetically non responsive strain. No clear-cut segregation of skin AHH inducibility levels is found among the offspring from the back-cross between (C57BL/6J X DBA/2J)F1 and non-inducible DBA/2J mice. PMID- 7470384 TI - In vitro osteolytic activity of human breast carcinoma tissue and prognosis. PMID- 7470385 TI - A study of histological features distinguishing chordoma from chondrosarcoma. PMID- 7470387 TI - A case-control study of hair-dye use and cancers of various sites. PMID- 7470386 TI - A cohort study of mortality from cancer of the prostate in Catholic priests. PMID- 7470388 TI - Different response of ENU-exposed and unexposed rat brain cells to cholera toxin at early passages in culture. PMID- 7470389 TI - Response of Chinese hamster ovary cells to anticancer drugs under aerobic and hypoxic conditions. PMID- 7470390 TI - Membrane proteins synthesized by human reticulocytes and their precursors. AB - Membrane protein synthesis in human immature erythroid cells was studied by incubating the cells with 35S-methionine in vitro. The radioactive precursor amino acid was incorporated into membrane protein in a linear fashion for approximately 60 min, after which there was only a slight increase in incorporation. Intracellular protein synthesis, in contrast, was linear for periods up to 2 h. Analysis of isolated membranes by polyacrylamide gel electrophoresis in sodium dodecyl sulphate showed that peripheral blood reticulocytes synthesized two proteins in the 4.5 region (MW=50-60000 D) and a third protein coinciding with band 6 (glyceraldehyde-3-phosphate dehydrogenase). Separation of reticulocytes into different age groups on stractan II gradients showed more immature reticulocytes synthesize a wider range of membrane proteins, extending from bands 4.1 to 8. When nucleated red cells were present in the incubations, synthesis of band 3 was also observed. Earlier erythroid precursor present in erythropoietic BFU-E cultures synthesized spectrin in addition to the other membrane proteins. The data indicate that human red cell membrane protein synthesis follows a programmed pattern and that as the erythroid elements mature they lose the capacity to synthesize certain membrane proteins. PMID- 7470391 TI - Effect of oxymetholone on human cyclic haematopoiesis. AB - A 62-year-old woman had recurrent fluctuations of neutrophil, monocyte, reticulocyte and platelet counts with a cycling time of 38.5 d. Periodic fluctuations of granulo-, erythro- and megakaryopoiesis were also demonstrated in her bone-marrow. Oxymetholone reduced the magnitude of the profound cyclic fluctuations in all these cell lines. During treatment a reduced number of in vitro CFU-C of the progenitor myeloid series from the bone marrow was observed. Using the patient leucocytes as feeders for normal control marrow cells, indication was found for a fluctuation in colony stimulating activity in vitro, rising above normal at the nadir of the WBC. PMID- 7470392 TI - Effects of shortened erythropoietin exposure on sheep marrow cultures. AB - To determine whether erythropoietin (Ep) is required only to trigger, or also to sustain the growth of erythroid progenitor (CFU-E) in vitro, the effect of limiting the time of exposure of sheep bone marrow CFU-E to Ep on erythroid colony (EC) formation was examined. Restricting the in vitro exposure of CFU-E to 2-24 h, either by brief preincubation with Ep or by neutralization of Ep present in culture with anti-Ep, resulted in a 70-90% reduction of EC formation, and the colonies that were produced were of a small size. In contrast, no inhibition of EC size or number occurred when CFU-E were exposed to EP for more than 24 h. These results suggest that Ep also modulates the proliferative events subsequent to the initial triggering of CFU-E which lead to the formation of erythrocytes. PMID- 7470393 TI - Hereditary antithrombin III deficiency in an English family. AB - A family including four members with decreased antithrombin III (ATIII) measured by a biological and an immunological method is described. Immunologically measured levels of other coagulation inhibitors were normal. In the antithrombin III deficient family members there was a history of thrombotic disease. Turnover of 125I-antithrombin III in a deficient individual showed the lower plasma levels probably to be due to a deficiency in the rate of synthesis rather than to an increased catabolic rate. PMID- 7470394 TI - Hereditary bleeding disorder due to a primary defect in platelet release reaction. AB - A large family with a hereditary bleeding disorder was investigated. Easy bruising, epistaxis and menorrhagia were noted in seven members of three generations and at least one member in each generation was affected. Platelet function abnormalities were characterized by reduced 14C-serotonin release, absent second wave aggregation in response to ADP or epinephrine and reduced aggregation in response to collagen. Bleeding time was prolonged in three individuals and platelet factor 3 availability was abnormal in four. Platelet count, morphology, adhesiveness and clot retraction were normal in all. Platelet ADP and ATP as well as ATP to ADP ratio were normal. This family probably represents the first documented instance of hereditary platelet primary release disorder. To elucidate the pathogenetic mechanism, further functional studies were performed. No appreciable shape change, 14C-serotonin release of aggregation was observed when the propositus' platelets were stimulated with sodium arachidonate or a PGH2 analogue. By contrast, platelets responded normally to ionophore A23187, thrombin and ristocetin. The findings indicate that the hereditary primary release disorder is probably due to a reduced thromboxane A2 production secondary to thromboxane synthetase deficiency. Alternatively, it may be due to platelet membrane abnormalities which render platelets unresponsive to thromboxane A2. PMID- 7470395 TI - An evaluation of chromogenic substrates in the control of oral anticoagulant therapy. AB - A comparison has been made between the prothrombin time test using British Comparative Thromboplastin (BCT) and four modified amidolytic assays in the assessment of laboratory control of oral anticoagulant administration in short term and long-term patients. The results were also correlated with the APTT and specific assays for factors VII and X based on established coagulation techniques. The chromogenic substrate assays did not give as close agreement with the BCT as the clotting assays for factors VII and X. A combination of the amidolytic factor II and X results did not improve the correlation. Agreement with the BCT was better in long-term patients with the substrates but there was no obvious advantage between the factor II and X amidolytic methods in this group. The development of a specific amidolytic assay for factor VII should be considered based on the evidence of this study although its reliability for oral anticoagulant control would need to be validated by clinical trial. In the interim, it may be anticipated that thromboplastins showing less sensitivity than BCT to factor VII would give a better correlation with the amidolytic assays. PMID- 7470396 TI - Difficulties in antenatal diagnosis of inherited haemoglobinopathies: gamma-chain variants. PMID- 7470397 TI - Eosinophilia in leukaemia. PMID- 7470398 TI - Relative importance of cigarette smoking in occupational lung disease. AB - Since 1900 respiratory disease has remained a constant serious cause of chronic ill health and premature death in Britain. The falling importance of tuberculosis and pneumonia has been off-set by the rise in lung cancer. Bronchitis morbidity and mortality have fallen only slightly since 1935. To produce any real improvement in the future existing information as to cause must be studied. The relative contribution of occupational exposure is compared with the importance of cigarette smoking. Relevant information is scanty and has been produced to emphasise the existence of occupational diseases rather than assess their importance to the community as whole. In Britain the evidence is that within the coal mining and iron and steel industries conditions are now such that dust exposure contributes little to the morbidity or mortality compared with the workers' smoking habits. Similar results have been shown by a cross-sectional survey of many dusty occupations in Western Germany. Only in the disappearing Welsh slate industry has dust disease been at least as important as smoking. Until the current regulations were introduced conditions existed among asbestos workers such that the combined effect of cigarette smoking and dust exposure led to a loss of life expectation of over 10 years in moderate smokers. Since the new regulations were introduced the risk for asbestos workers should approximate to that for other industrial workers. While control of occupational exposure to respiratory hazards remains important, a far greater improvement to respiratory health would be produced by controlling tobacco smoking. PMID- 7470399 TI - An experimental study on the combined effects of n-hexane and toluene on the peripheral nerve of the rat. AB - An electrophysiological study was undertaken to determine whether toluene affected the neurotoxicity of n-hexane. Separate groups of eight rats were exposed to 1000 ppm n-hexane, 1000 ppm toulene, 1000 ppm n-hexane plus 1000 ppm toluene, of fresh air in an exposure chamber for 12 hours a day for 16 weeks. The body weight, MCV, DL, MNCVs were measured before exposure, after four, eight 12, and 16 weeks exposure; and four weeks after exposure was discontinued. Exposure to 1000 ppm n-hexane considerably impaired the function of the peripheral nerve, but exposure to a mixture of 1000 ppm n-hexane plus 1000 ppm toluene resulted in only slight impairment; 1000 ppm toluene had little effect. These results strongly suggest that toluene decreases the toxic effects of n-hexane on the peripheral nerve. PMID- 7470400 TI - Urinary excretion of the metabolites of n-hexane and its isomers during occupational exposure. AB - Environmental exposure to commercial hexane (n-hexane, 2-methylpentane, and 3 methylpentane) was tested in several work places in five shoe factories by taking three grap-air samples during the afternoon shift. Individual exposure ranges were 32-500 mg/m3 for n-hexane, 11-250 mg/m3 for 2-methylpentane, and 10-204 mg/m3 for 3-methylpentane. The metabolites of commercial hexane in the urine of 41 workers were measured at the end of the work shift. 2-Hexanol, 2,5 hexanedione, 2,5-dimethylfuran, and gamma-valerolactone were found as n-hexane metabolites and 2-methyl-2-pentanol and 3-methyl-2-pentanol as 2-methylpentane and 3-methylpentane metabolites. The presence of metabolites in the urine was correlated with occupational exposure to solvents. n-Hexane exposure was correlated more positively with 2-hexanol and 2,5-hexanedione than with 2,5 dimethylfuran and gamma-valerolactone. A good correlation was also found between total n-hexane metabolites and n-hexane exposure. 2-Methyl-2-pentanol and 3 methyl-2-pentanol were highly correlated with 2-methylpentane and 3-methylpentane exposure. The results suggest that the urinary excretion of hexane metabolites may be used for monitoring occupational exposure to n-hexane and its isomers. PMID- 7470401 TI - Soft-tissue sarcomas and exposure to chemical substances: a case-referent study. AB - In 1977 several patients were seen with soft-tissue sarcomas and previous exposure to phenoxy acids. This clinical observation resulted in a cases-referent (case-control) study being undertaken which showed that exposure to phenoxy acids or chlorophenols, which are chemically related, gave a roughly six-fold increase in the risk for this type of tumour. A further case-referent study of soft-tissue sarcomas has now been performed to confirm these earlier findings and also to obtain further information on the effects of different phenoxy acids. This new investigation gave an increase of the same magnitude in the risk for soft-tissue sarcomas after exposure to phenoxy acids or chlorophenols, but this risk related also to exposure to phenoxy acids free from impurities, such as polychlorinated dibenzodioxins and dibenzofurans. PMID- 7470402 TI - Lung function of office workers exposed to humidifier fever antigen. AB - Office workers who became sensitised to antigens derived from humidifier sludge developed episodes of fever, malaise, and other symptoms, including polyuria and mild chest tightness. The episodes usually occurred on a Monday evening and were to some extent dose-related. Lung function was assessed over a day shift on two occasions, including one after which almost all the susceptible subjects developed symptoms. The symptoms were preceded by a 6% reduction in forced expiratory volume and vital capacity, a corresponding increase in residual volume, and a reduction in flow rate after 75% vital capacity had been expired. There were no changes in peak expiratory flow, forced expiratory flow at 50% of vital capacity, or transfer factor. In some subjects the transfer factor was apparently reduced 36 hours later, but for this there may have been another explanation. The physiological features were considered to reflect narrowing of small airways in the lung. The changes, however, were minimal and not the main cause of the symptoms. A feature of the episode was the severity of the constitutional symptoms despite the low airborne dust levels. PMID- 7470403 TI - Laboratory animal allergy: a clinical survey of an exposed population. AB - A clinical survey of workers exposed to laboratory animals in a pharmaceutical company was designed to discover the prevalence and severity of symptoms of laboratory animal allergy (LAA). The overall prevalence of the condition was 30%, and two distinguishable LAA syndromes, termed regional and progressive LAA, were found. The first is characterised by rhinitis with negative skin prick tests. The second consists of rhinitis leading progressively to asthma with positive prick tests. Prick tests were useful diagnostically only in the latter. Atopes were shown not to be at special risk of developing LAA, but if they did so were more likely to progress to asthma. The implications for selection and management policy are outlined and specific measures for the further study of LAA are proposed. PMID- 7470404 TI - Intrabronchial instillation of paraquat in rats: lung morphology and retention study. AB - Various amounts of paraquat (10(-5) to 10(-12) g) in 0.1 ml saline were instilled directly into the left bronchus of male adult rats. Gravimetric, macroscopic, and microscopic studies on the left lobe of the lung showed that 10(-5) g of paraquat produced lung oedema and macroscopic lesions two and 14 days after doing. The pathology of the lung was similar to that seen after systemic poisoning. When 10( 6) g of paraquat was instilled, some animals developed lung oedema and macroscopic lesions. Microscopic examination showed subtle changes in the parenchyma of the lung. With amounts of paraquat equal to or less than 10(-7) g (doses as little as 10(-12) g were used), no changes in the lung were seen. This is contrary to published accounts in which amounts as low as 10(-12) g (1 Pg) were reported to cause acute damage to the rabbit lung. When 3H paraquat was instilled into the left lobe (doses of 10(-5) to 10(-10) g were used), the loss of paraquat from the lung was biphasic. The initial half-life was less than one hour. The secondary phase obeyed first-order kinetics, and the half-life was dependent on the dose of paraquat instilled. This half-life was as short as 11 hours when 10(-5) g paraquat was instilled and was 76 hours after the instillation of 10(-10) g paraquat. The decrease in the half-life of the secondary phase with increasing doses of paraquat is possibly associated with the production of oedema or lung cell damage, or both. After the instillation of 10( 8) g 3H paraquat, the initial half-life was less than 15 minutes, and paraquat was detected in the urine and plasma at that time. This suggests that 50% of the instilled paraquat was rapidly absorbed from the lung into the plasma. PMID- 7470405 TI - Scanning electron-microscopic and light-optic investigations of erythrocytes in toxic anaemia. AB - Detailed assessment of the peculiarities of erythrocyte morphology by applying scanning electron microscopy and light-optics methods gives valuable information on the age characteristics, functional failures of the erythrocyte, pathogenesis of anaemia, and also diagnosis. In toxic anaemia caused by the chronic effect of lead and chlorobenzene there is accumulation of aging erythrocytes in blood that are spherical with rough and folded surfaces, fragmented with protuberances, processes, crypt-like hollows, and holes. In lead intoxication there is increased destruction of erythrocytes; anaemia caused by chlorobenzene is mainly due to a decreased entry of the young forms of the erythrocyte into the blood. Light optics investigation combined with scanning electron microscopy allows a quantitative calculation of erythrocytes with abnormalities of form and surfaces. This may be used as an additional diagnostic test as part of the haematological examination of patients with anaemia. PMID- 7470406 TI - Chronic cadmium poisoning in a pigment manufacturing plant. AB - When the working environment in a small cadmium-pigment manufacturing plant was investigated cadmium concentrations in respirable dust were found to be considerably above the hygiene standard recommended by the British Occupational Hygiene Society. Cadmium concentrations in blood and urine of exposed workers, both present and past employees, were determined. The six men who had worked in the production plant for seven years or more all showed signs of renal tubular damage. In addition two of these men exhibited exertional dyspnoea, and respiratory function tests showed evidence of pulmonary dysfunction. A third man showed moderate airways obstruction with mild symptoms. On the basis of the results of the investigations on individuals in this study, it is suggested that urinary cadmium concentrations should be kept below 15 microgram/day to avoid the possibility of renal damage, and that the insoluble respirable fraction of cadmium dust should not be regraded as merely nuisance dust. PMID- 7470407 TI - Locally induced digital vasospasm detected by delayed rewarming in Raynaud's phenomenon of occupational origin. AB - The rewarming time of fingers after hand cooling to 10 degrees during ischaemia was significantly prolonged in all subjects with Raynaud's phenomenon of occupational origin compared with normal reaction and patients with primary Raynaud's phenomenon. The temperature is faster and easier to record than pressure or pulse volume. Population studies are in progress with the described technique. PMID- 7470408 TI - Induction of microsomal drug metabolism in man and in the rat by exposure to petroleum. AB - To determine the effect of petroleum exposure on the activity of hepatic mixed function oxidase enzymes, salivary elimination kinetics of antipyrine were determined in 19 petrol station attendants and compared with 19 controls. Antipyrine half life in petrol station attendants was shorter than in controls. Microsomal preparations (10 000 x g supernatants) were prepared from six male Porton rats exposed to petrol vapour (5 ppm at an air flow rate of 41/min for eight hours a day for three weeks) and six control rats maintained under the same conditions without exposure to petrol vapour. The rates of oxidative metabolism of antipyrine, aminopyrine, ethylmorphine, aniline, and benzo(a)pyrene were all increased by more than 45% in the petrol-exposed rats. The results indicate that petrol vapour is a moderately potent inducer of mixed function oxidase activity in rats, and that occupational exposure to petroleum may result in enhanced microsomal drug metabolism. PMID- 7470409 TI - Correlation of urinary thioethers with chemical exposure in a rubber plant. AB - Exposure to potentially alkylating in a rubber factory was measured by determining thioether concentrations in urine samples collected at the end of work on a Friday. The study population consisted of the total work force (113) in the production departments of a factory, and office clerks (111) in two factories of the same company. The highest excretion of thioethers was detected in female workers in the belt department who were exposed through the palmar skin. High excretion was also found in workers exposed to air-borne contaminants in the calender department and in workers in the raw material stores and chemical mixing sections. Lower values were found in this dispatching station and in the contaminant-free offices. Urinary thioether determination appears to be a reliable and easy method of assessing the exposure to certain rubber chemicals. PMID- 7470410 TI - Epidemiological evidence for two components of cervical cancer. AB - Incidence rates for cervical cancer were analyzed for Finland in 1953 to 1974. The age-specific incidence rates fit an age-incidence curve based on the assumption of two epidemiological components. The time trends give support to the hypothesis that one of the components occurs at a lower age and is preceded by a carcinoma in situ stage. The risk of cervical cancer originating from the component occurring in younger women showed a decreasing trend which correlated with the organized mass screening programme. The other component occurred later in life and was not influenced by the screening programme. It was estimated that the proportion of the cases in the earlier category decreased from 53% in the 1950s to 17% in 1973 to 1974 of all cases of cervical cancer diagnosed in the Finnish population. PMID- 7470411 TI - The obstetric outcome of teenage pregnancy. AB - A retrospective case record analysis of the obstetric outcome in 715 consecutive teenage primigravidae was performed. They were late in booking for hospital confinement and were frequently unsure of their expected date of delivery; 82% were unmarried before conception. When compared with primigravidae aged 20 to 24 years, anaemia was the only antenatal complication that was significantly increased. There was no evidence of an increased obstetric risk in the teenage women as a group but those who remained single showed significantly higher rates of premature labour and perinatal mortality when compared with the married women and appear to constitute an at risk group. PMID- 7470412 TI - The merits of special antenatal care for twin pregnancies. AB - A combined series of 101 twin pregnancies for whom routine hospital rest during the last trimester was replaced by intensified antenatal care in a special twins clinic was studied. Perinatal mortality and morbidity was similar to that found in a comparison group of 137 twin pregnancies under the care of consultants and not referred to the twins clinic. Routine cervical assessments and uterine activity measurements were unhelpful in predicting premature delivery. Urinary oestrogens and ultrasonic measurements of fetal biparietal diameter were of little or no value in predicting weight for gestational age but the ultrasonic measurement of abdominal circumference provided limited information on fetal growth. The contribution of a placebo effect to the results of patients receiving more personalised care cannot be discounted. PMID- 7470413 TI - A comparison of oestradiol and prostaglandin E2 for ripening the cervix. AB - A study was conducted to compare the cervical ripening effect of extra-amniotic prostaglandin E2 with that of extra-amniotic oestradiol valerate in preparation for induction of labour. Sixty primigravidae with an unripe cervix received one or other agent suspended in a viscous gel. The degree of cervical ripening was similar in both groups but more patients treated with prostaglandins became established in labour before planned amniotomy, and poor progress in labour was less common in this group. PMID- 7470414 TI - Changes in serum bile acid concentrations during normal pregnancy, in patients with intrahepatic cholestasis of pregnancy and in pregnant women with itching. AB - Two primary bile acids, cholic acid (CA) and chenodeoxycholic acid (CDCA), and one secondary bile acid, deoxycholic acid (DCA), were measured by radioimmunoassay in pregnancy serum from 30 healthy women, 49 women with itching and 45 with intrahepatic cholestasis of pregnancy. All subjects were studied serially from between 16 and 20 weeks gestation until 35-60 days post partum. In healthy pregnant women, serum CA and DCA levels did not change significantly at any time. The mean CDCA level rose significantly towards term. In women with intrahepatic cholestasis, serum levels of CA and CDC were increased ten- and five fold, respectively, at the time of appearance of clinical symptoms and the CA/CDCA ratio rose from 1/1 to 2/1; there was also a moderate increase in the serum concentration of DCA. In 4 of 8 women studied prospectively an increase in serum bile acid levels preceded the appearance of symptoms or other laboratory evidence of intrahepatic cholestasis. Nine of the women with itching with normal routine liver function test results had increases in serum CA and CDCA concentrations suggesting mild cholestasis. PMID- 7470415 TI - Assessment of gestation by ultrasound in various stages of pregnancy in infants differing in size and ethnic origin. AB - A clinical assessment of gestational age after birth has been compared with the ultrasound measurement in utero at various stages of pregnancy in a population of mixed socio-economic status and racial origin. In infants who were small-for dates or large-for-dates at birth, accurate prediction from ultrasound measurement could only be achieved if the crown rump length or the biparietal diameter was measured before 20 weeks gestation. At this time, the infants destined to be small-for-dates or large-for-dates at birth show no growth retardation or acceleration. There is also no influence of racial origin on the measurements. PMID- 7470416 TI - Ultrasound demonstration of an unusual fetal growth pattern in indians. AB - An ultrasound study of fetal growth patterns in women of Indian origin showed that the mean abdominal circumference measurements throughout pregnancy were significantly smaller than measurements obtained in white European patients. The mean birth weight of the Indian babies was 340 g less than that of the controls and Indian patients also exhibited a significantly shorter duration of gestation. There was no significant difference between the biparietal diameter measurements of European and Indian women throughout pregnancy. PMID- 7470417 TI - The prediction of fetal growth retardation in twin pregnancy. AB - A total of 132 twin pregnancies first seen at less than 36 weeks gestation were studied prospectively to determine the epidemiological and anthropomorphic factors associated with single or dual fetal growth retardation; the clinical, biochemical and ultrasound serial measurements that are predictive of single or dual fetal growth retardation, and to design scoring systems for the prediction of fetal growth retardation. A number of factors were associated with an increased risk of fetal growth retardation, but the highest risk was present when there was an abnormality in both plasma oestriol and placental lactogen, and the biparietal diameter growth rates were divergent. Scoring systems were derived using multi-variant discriminant analysis for three clinical situations: the patient seen for the first or second time; where only clinical facilities exist and the patient was seen on three or more occasions; and where facilities exist for the biochemical tests of placental function and ultrasonic measurement of the biparietal diameters. The scoring system for the last situation had the highest predictive rate and the lowest false positive rate. PMID- 7470418 TI - Prostaglandin E and F concentrations in placentae of normal, hypertensive and pre eclamptic patients. AB - The prostaglandins E and F were measured by radioimmunoassay in placentae from 23 normotensive, 25 hypertensive and 13 pre-eclamptic patients. The prostaglandin E levels were respectively (pg/mg +/- SEM) 16.34 +/- 2.06, 16.75 +/- 2.52 and 12.10 +/- 3.63. The prostaglandin F levels were 2.43 +/- 0.41, 2.70 +/- 0.42 and 2.06 +/- 0.31. These prostaglandin concentrations did not differ significantly. Local ischaemia may, therefore, be explained by altered responsiveness to these prostaglandins or to changes in other prostaglandins (such as the vasodilator PGI2) or other vasoactive agents. PMID- 7470419 TI - Origin of amniotic fluid cells in mid-trimester pregnancies. AB - Amniotic fluid cells provide important information about the fetus. Studies of the surface ultrastructure of such cells identified four cell populations in mid gestational fluids. A parallel study of tissue surfaces in contact with or communicating with the amniotic fluid showed that some tissues made no contribution to the cell populations, while others contributed extensively. Two principal cell shedding mechanisms were found in mid-trimester--exfoliation of whole cells and detachment of cellular fragments. The latter mechanism was observed from skin and umbilical cords during a limited period in mid-pregnancy. The majority of cells were derived from exfoliation of squamous epithelia. PMID- 7470420 TI - Morphologic characteristics of the normal term human trophoblast maintained in prolonged in vitro cultures. AB - The development of a new technique for prolonged in vitro culture of normal human term trophoblast without stromal growth has facilitated the study of the morphologic characteristics of trophoblast in culture. Cytotrophoblast (epithelioid) cells were observed along with subsequent formation of syncytiotrophoblast in vitro. Hormonal assessment of hPL production has confirmed the viability and biologic specificity of the tissue. The morphologic characteristics of normal human term trophoblast maintained in vitro include projections, circular growths and syncytial sprouts. Growth patterns of trophoblast in vitro were similar to those described in vivo. PMID- 7470421 TI - The maintenance of human amniotic membranes in culture. AB - A culture systems is described for the long-term maintenance of human amnion in vitro. This has been developed because of the need for an amnion organ culture to study immunological aspects of this extra-embryonic membrane as well as the increasing surgical use of amnion in wound healing. The intact amnion can be maintained for 2-3 weeks with 50-90% viability as determined by morphological and trypan-blue extraction techniques. This puts forward the feasability of developing an amnion bank for clinical investigations of wound healing as well as the possibility of in vitro studies of this membrane in the materno-fetal relationship in human pregnancy. PMID- 7470422 TI - The effect of pre-operative povidone iodine vaginal pessaries on vault infections after hysterectomy. AB - The frequency of clinical infection of the vaginal vault after hysterectomy was compared in two groups of patients, a study group treated with povidone-iodine vaginal pessaries for 36 hours pre-operatively and prepared with povidone-iodine preparations at the time of operation, and a control group given no preliminary treatment and prepared with chlorhexidine preparations at the time of operation. No significant difference was demonstrated between the two groups. Vault infection occurred in 32% of the entire series of patients (severe infection 14%) and infection correlated with the presence of pathogenic bacteria in the vagina. No significant differences in bacterial colonisation of the vagina between the two groups were demonstrated at any stage of the investigation. Preliminary preparation of the vagina with povidone-iodine pessaries is not recommended. PMID- 7470423 TI - Reversal of female sterilization. AB - Reversal of sterilization was attempted in 22 women without the use of an operating microscope. Fifty per cent became pregnant. Success was influenced by the site and extent of the tubal damage resulting from the sterilization procedure, mechanical occlusive sterilization techniques offering the best change of a successful reversal. PMID- 7470424 TI - A case of clear cell adenocarcinoma of the vagina in pregnancy. AB - The case is described of a 20 year old pregnant woman with clear cell adenocarcinoma of the vagina who had been exposed to diethyl stilboestrol in utero. This is the second of three cases known to have occurred in Britain. The importance of radical treatment is discussed. Neither cervical nor vaginal intra epithelial neoplasia were seen histologically, although squamous metaplasia was widespread, and in some areas was immature and showed atypical features which could be misinterpreted as intra-epithelial neoplasia. An area of atypical adenosis was noticed, a hitherto rarely observed feature. PMID- 7470425 TI - Pregnancy in women after repair of bladder exstrophy. Two case reports. AB - Improved techniques in urinary diversion operations have enabled a large number of patients with bladder exstrophy to achieve urinary control. They can live fairly normal lives and some have achieved pregnancy. Case reports of such exist in the literature. Two more cases are reported in this paper, the pregnancies and their outcome are discussed. PMID- 7470426 TI - Degradation of maize protein in rumen contents. Influence of ammonia concentration. AB - 1. The influence of ammonia concentration on the distribution of nitrogen derived from opaque-2 maize uniformly-labelled with 15N has been investigated during short-term in vitro incubation of bovine rumen contents. 2. Less 15N derived from maize was found in the non-protein-N (NPN) fraction during incubation without added NH3 than with added NH3, due entirely to differences in the amount of N derived from maize in the NH3 fraction. 3. From calculations based on the transfer of N derived from maize to the NPN pool and to a bacterial fraction, it was concluded that degradation of maize protein was not influenced by NH3 concentration within the examined limits. 4. The decrease in relative amount of N derived from maize in the NH3 fraction at low concentrations of NH3, together with evidence for an increased fractional turnover rate of NH3-N suggests that a deficient supply of NH3 is compensated for by increased catabolism of nitrogenous compounds derived from the rumen micro-organisms. PMID- 7470427 TI - Selenium metabolism in the dairy cow: the influence of the liver and the effect of the form of Se salt. AB - 1. Six adult Friesian cows were given 75Se as either 75SeO32- or 75SeO42- intravenously. Five of the cows had cannulas in an hepatic vein, the portal vein and one carotid artery to enable the uptake of 75Se by the liver to be measured. Radioactive balance studies were carried out on two of the cows given 75SeO32- and two given 75SeO42-. A seventh cow was given an oral dose of 75Se-labelled barley and the excretion of 75Se in faeces, urine and milk was measured for 14 d. 2. After the injection of 75SeO32- plasma 75Se concentration decreased during the first 30 min with a mean half-life (t 1/2) of 15.6 min. From 30 to 60 min after dosing the concentration of radioactivity increased to reach approximately 50% of the level present 2 min after dosing. Following the injection of 75SeO42- the 75Se was cleared with a mean t 1/2 of 28.5 min during the first 30 min and plasma radioactivity increased only slightly during the next 30 min. 3. during the phase of rapid clearance of 75Se after the injection of 75SeO32- the hepatic venous 75Se concentration was approximately 5% lower than portal venous 75Se concentration. During the period when plasma 75Se activity was increasing the activity in hepatic venous plasma was 3% greater than portal activity. Of the 75Se cleared from plasma after injecting 75SeO32- 40% was calculated to be removed by the liver. 4. After intravenous dosing with 75SeO32- or 75SeO42- approximately 9.5 and 17.0% respectively of the dose injected was excreted in faeces and 10% in urine within 14 d. Almost three times as much 75Se was excreted in urine and 3.5 times as much in faeces during the first 24 h after dosing with 75SeO42- as after 75SeO32-. PMID- 7470428 TI - Effect of zinc deficiency on appetite and plasma amino acid concentrations in the rat. AB - 1. Levels of zinc in liver and plasma of the Zn-depleted rats fluctuated with the feeding cycle and were significantly higher at the bottom than at the top of the cycle. As Zn deficiency became more severe fluctuations in plasma Zn diminished. Concentrations of Zn in liver, in contrast to levels in plasma and femur, were not markedly lowered by day 15. 2. In contrast to ad lib-fed (AL) and overnight fasted (OF) controls, some pair-fed (PF) controls had elevated levels of Zn in liver and plasma. 3. Intakes of water and food were significantly correlated in Zn-deficient rats. Packed cell volumes were significantly higher for Zn-depleted than for AL and PF rats. 4. Food intakes and plasma glucose concentrations were related in AL, OF and PF control rats but not in Zn-deficient rats. 5. At day 15 of Zn deficiency the order of total plasma amino acid concentrations in the groups of rats was AL greater than Zn-deficient greater than OF greater than PF. Many of the differences between the AL and OF groups for individual plasma amino acids also appeared in the Zn-deficient group at the top and bottom of the feeding cycle. Differences in individual amino acid concentrations at the top and bottom of the feeding cycle tended to be opposite in the PF and the Zn-deficient group. Levels of tyrosine and tryptophan in plasma were correlated (P less than 0.05) with the cyclic feeding pattern of the Zn-deficient group; however, the ratios tryptophan or tyrosine: sum of other large neutral amino acids did not correlate significantly with the eating habits of Zn-deficient rats. PMID- 7470429 TI - Studies on the nutrition of salmonid fish. The magnesium requirement of rainbow trout (Salmo gairdneri). AB - 1. Rainbow trout (Salmo gairdneri) of mean initial weight 35 g were given one of five experimental diets for 20 weeks. The diets contained (g/kg dry diet) 15 calcium, 10 phosphorus and graded levels of magnesium from 0.04 (diet no. 1) to 1.0 (diet no. 5). In a second experiment rainbow trout of mean initial weight 16 g were given one of six experimental diets for 20 weeks. The diets contained (g/kg dry diet): Ca (40), P (30) and levels of Mg from 0.06 (diet no. 6) to 2.0 (diet no. 11). 2. In both experiments weight gains were lowest in those trout given diets containing the basal levels of Mg (diet no. 1 and diet no. 6) but increased with increasing dietary Mg concentration. In neither experiment was there any further increase in weight gain once the Mg concentration reached 0.25 0.5 g/kg dry diet; weight gain reached a plateau at this dietary MG level. 3. The following trends occurred in serum electrolyte concentrations as dietary Mg increased. Mg increased in both experiments, in Expt 2 it reached a maximum of 1 mmol/l when the diet contained 0.5 g Mg/kg and did not increase further; sodium was positively correlated in both experiments; potassium decreased and in Expt 2 reached a plateau minimum of 1.7 mmol/l at a dietary Mg concentration of 0.5 g/kg; Ca and P altered little in either experiment. 4. In both experiments renal Ca concentrations were greatly increased in trout given diets lacking supplementary Mg; they fell to low levels (3-5 mmol/kg) when diets contained 0.15 g Mg/kg or more. Renal K and P concentrations were negatively correlated with dietary Mg in Expt 2; other electrolytes measured were not altered in concentration by the treatments used. 5. Extracellular fluid volume (ECFV) of muscle was negatively correlated with dietary Mg. In Expt 2 it reached a minimal or normal value at 0.5 g Mg/kg diet and did not decrease further. Muscle Mg concentration increased with diet Mg in both experiments and muscle K concentration was also correlated with diet Mg in Expt 2. These changes were related to the shift in muscle water. In Expt 1, P concentration was decreased with increasing diet Mg but in Expt 2 its concentration increased, these changes may have been connected with the three-fold difference in dietary P in the two experiments. 6. By contrast with skeletal muscle, Mg levels in cardiac muscle increased at low dietary Mg intakes. 7. Concentrations of electrolytes in liver did not alter with the dietary treatments used. 8. The results show that Mg requirement of rainbow trout is met by a diet containing 0.5 gMg/kg diet. PMID- 7470430 TI - Changes in the concentrations of the minor constituents of goat's milk during starvation and on refeeding of the lactating animal and their relationship to mammary gland metabolism. AB - 1. Changes in the concentrations of the minor constituents of goat's milk were observed during 48 h starvation and on refeeding. 2. The concentrations of hexose phosphate and UDP-hexoses increased during starvation and decreased on refeeding. 3. The concentrations of phosphoenolpyruvate and glycerate 3-phosphate decreased during starvation and increased on refeeding. 4. Isocitrate:2-oxoglutarate increased during starvation and decreased on refeeding. 5. Changes in the minor constituents of milk can be explained in terms of the metabolic changes occurring in the mammary gland during starvation. It is proposed that changes in the concentrations of these metabolites in milk reflect changes in their concentrations in the cytosol or Golgi vesicles of the mammary gland. PMID- 7470431 TI - Diabolic acids: occurrence and identification in natural products and their metabolism by simple-stomached and ruminant animals. AB - 1. A method for gas-liquid chromatographic analysis of the dimethyl esters of a homologous series of diabolic acids is described. 2. Only when combined with low resolution mass spectrometry could the method be used to show unequivocally the existence of diabolic acids in natural products. 3. Diabolic acids occurred naturally in rumen and faecal lipids of all ruminants examined but could not be detected in a number of ruminant tissue samples. 4. Infused 14C-labelled (C16:0)2 diabolic acid was poorly absorbed by both simple-stomached and ruminant animals. Oxidation was more extensive in the ruminant, but there was no evidence of deposition of diabolic acid in the tissues of either animal. PMID- 7470432 TI - The digestibility of amino acids in the small intestine of the sheep. AB - 1. The digestibilities of microbial and food proteins in the small intestine were studied in three sheep fitted with re-entrant cannulas in the proximal duodenum and terminal ileum. 2. The quantities of microbial and food proteins at the small intestine were varied by infusion of a microbial isolate or by dietary manipulation and the balance of amino acids along the small intestine was determined. 3. A mean value of 0.69 for the apparent digestibility and 0.86 for the true digestibility of total amino acids was obtained. 4. From the composition of digesta at the duodenum the daily flows of microbial and food proteins were estimated. Their true digestibilities in the small intestine were calculated by regression and found to be: microbial protein 0.87 and food protein 0.82. The mean endogenous loss of amino acids secreted into the small intestine was estimated to be 13.3 g/d. PMID- 7470433 TI - The effect of amino acid imbalance on nitrogen retention (biological value) in rats. AB - 1. Three series of nitrogen balance experiments were performed with growing rats to test the effect of amino acid imbalance on protein utilization. In Expt 1 egg protein was fed as a basal diet and supplemented with one amino acid at a time in the amounts originally found in egg protein. In Expt 2 the procedure was repeated with a barley diet, and in Expt 3 egg and potato proteins were fed together in various combinations. 2. Doubling the single amino acids, especially arginine, in an egg-protein diet reduced biological value (BV) markedly. Also the branched chain and the basic amino acids reduced BV significantly. In the barley-based diets negative effects were also observed when the concentration of the single amino acid was doubled. However, it was assumed that the observed deleterious effects of amino acid excess were partly due to an exaggerated lysine deficiency in the barley protein when non-limiting amino acid excess were added. 3. The results with various combinations of egg and potato proteins showed that as potato protein was increased true protein digestibility decreased linearly, whereas BV decreased curvilinearly. Maximum protein utilization was obtained with egg protein alone. PMID- 7470434 TI - Factors affecting the voluntary intake of food by sheep. 3. The effect of intravenous infusions of gastrin, cholecystokinin and secretin on motility of the reticulo-rumen and intake. AB - 1. Sheep given ground and pelleted lucerne hay (Medicago sativa) ad lib. were infused intravenously with pentagastrin, secretin, cholecystokinin (CCK) and its analogues to assess their effects on motility of the reticulo-rumen and on food intake. In the latter experiments the animals were deprived of their diet for periods of up to 6 h to induce hunger and the infusions were made before and during 3-10 min periods of feeding. 2. Pentagastrin, and analogue of gastrin, depressed intake by 35-50% (P less than 0.05) when it was infused at 9 microgram/kg per h during 30 min of feeding. The threshold may however be below 1 microgram/kg per h as this dose decreased intake of 12-17%. The frequency of reticular contractions decreased by 13, 35, 39 and 44% when 1, 3, 9, and 27 microgram pentagastrin/kg per h respectively was infused (P less than 0.025). 3. Secretin depressed food intake 38% after 30 min (P less than 0.025) when 8 Clinical Units (CU)/kg per h was infused but the threshold could be less than this dose since 0.5 CU/kg per h depressed intake by 12%. Contraction amplitude but not frequency decreased at 8 CU/kg per h. 4. CCK produced a 39% decrease in intake during the first 10 min of feeding (P less than 0.05) and the threshold was between 5 and 15 Ivy Dog Units (IDU) or 425 and 1276 pmol/kg per h. The frequency of reticular contractions was not affected by 1.7 IDU/kg per h but it was depressed 21 and 63% by 5 and 15 IDU/kg per h. Octapeptide at 1.5 and 3 microgram (1312 and 2624 pmol) kg per h depressed intake by 11 and 43% respectively after 10 min (not significant) and 1.5 microgram/kg per h depressed motility by 39% (P less than 0.01). Ceruletide at 810 ng (599 pmol)/kg per h depressed intake by 31% (not significant) after 10 min and decreased motility by 52% (P less than 0.05). The threshold dose for ceruletide on intake appeared to be about 90 ng or 66 pmol/kg per h which is considerably less than that for CCK or octapeptide. 5. The biological significance of gastrointestinal hormones as signals of satiety in normal sheep is not known since doses of pentagastrin and CCK that suppressed intake also interfered quite markedly with motility. However there is good reason to suspect that elevated concentrations of gastrin and CCK in blood of parasitized sheep may account at least in part for their symptoms of rumen atony and reduced food intakes. PMID- 7470435 TI - Effect of different levels of dietary molybdenum on copper and Mo metabolism in sheep fed on high levels of Cu. AB - 1. Distribution of copper and molybdenum was followed in the body tissues of sheep fed on high levels of Cu (82 mg Cu/sheep per d), sulphur (3.77 g S/sheep per d) and different levels of Mo (0.6 20.8, 38.4 and 58.5 mg Mo/sheep per d). 2. Liver Cu content decreased as Mo intake increased from 0.6 to 38.4 mg/d, but increased again at high intakes of Mo. With an Mo intake of 58.5 mg/d, the Cu content of liver, kidney, lung, spleen and muscle tissue was significantly higher than with an intake of 20.8 mg Mo/d. The trend of increased Cu concentrations in kidneys and plasma was already evident at an Mo intake of 38.4 mg/sheep per d. 3. High positive correlations were observed between Cu and Mo in both the kidney cortex and medulla of the sheep at the two highest Mo treatments. 4. At constant S intake, Mo concentrations in the tissues tended to increase in proportion to Mo intakes. No indication of any detrimental effect due to the accumulation of Mo in the tissues was observed. 5. It was suggested that in the presence of an abundance of Mo, Cu and S, compounds containing these minerals in metabolically unavailable forms accumulate in the body, first in the kidneys, but eventually also in the other tissues of the sheep. PMID- 7470436 TI - A biochemical evaluation of the erythrocyte glutathione reductase (EC 1.6.4.2) test for riboflavin status. 1. Rate and specificity of response in acute deficiency. PMID- 7470437 TI - The effect of meal frequency and protein concentration on the composition of the weight lost by obese subjects. AB - 1. Nitrogen balance, weight loss and resting metabolic rate were measured in thirty-eight obese inpatients on 3.4 MJ (800 kcal)/d diets over 3 weeks. 2. All subjects were fed on 13% protein-energy in three meals/d for the first week. 3. In weeks 2 or 3, using a cross-over design, ten subjects were fed on 15 or 10% protein-energy as three meals/d; fourteen subjects were fed on five or one meal/d with 13% protein-energy; and fourteen subjects were fed on 15% protein-energy as five meals/d or 10% protein-energy as one meal/d. 4. N loss was least on the high protein week and frequent-meal week: the largest difference was found when these effects were combined (P less than 0.001). 5. When protein-energy was held constant at 13% N loss decreased significantly (P less than 0.01) between week 2 and 3, but when the protein-energy was manipulated there was no significant N conservation in the third week. This suggests that the protein:energy value is more important than meal frequency in the preservation of lean tissue. 6. Weight loss was also least on the 'high-protein' week and 'frequent-meal' week, but this result reached significance only when the effects were combined (P less than 0.05). 7. Resting metabolic rate decreased with time but was not significantly altered by the dietary regimens. 8. Therefore, during the first 3 weeks at an intake of 3.4 MJ/d, a diet with a high-protein concentration, fed as frequent small meals, is associated with better preservation of lean tissue than an isoenergetic diet with lower-protein concentration fed as fewer meals. There was no evidence that meal frequency or protein concentration affect the rate of fat loss. PMID- 7470438 TI - A biochemical evaluation of the erythrocyte glutathione reductase (EC 1.6.4.2) test for riboflavin status. 2. Dose-response relationships in chronic marginal deficiency. AB - 1. Chronic marginal riboflavin deficiency was induced in groups of weanling rats by feeding a deficient diet supplemented with 0, 0.5, 1.0 and 1.5 mg riboflavin/kg diet. Ad lib.- and pair-fed controls received 3.0 and 15 mg riboflavin/kg diet respectively. 2. Serial measurement of erythrocyte NAD(P)H2 glutathione oxidoreductase (glutathione reductase; EC 1.6.4.2) and its activation coefficient revealed that after 12 weeks a steady-state of deficiency had been reached following initial fluctuations in status; the animals were then killed, and their tissues analysed. 3. Food intake, growth rate and the appearance of pathological signs were directly proportional to riboflavin content; however relative liver weight was increased above control levels only in the most severely-deficient group, and anaemia was not detected in any group. 4. The activation coefficient of glutathione reductase in erythrocytes and liver was closely related to dietary riboflavin content; that of skin responded maximally even in the least-severely-depleted animals. 5. Hepatic and renal flavin contents were directly proportional to dietary riboflavin, FAD being conserved at the expense of riboflavin and FMN. ATP:riboflavin 5-phosphotransferase (flavokinase; EC 2.7.1.26) activity was reduced, even in the least-severely-deficient animals; ATP:FMN adenylyltransferase(FAD pyrophosphorylase; EC 2.7.7.2) was increased in liver, but only in the most-severely-deficient animals. 6. Hepatic succinate:(acceptor) oxidoreductase (succinate dehydrogenase; EC 1.3.99.1) activity fell sharply between 1.5 and 0.5 mg riboflavin/kg diet, producing an S shaped dose-response curve; it showed smaller or less specific changes in other tissues such as brain, skin and intestine. NADH:(acceptor) oxidoreductase (NADH dehydrogenase; EC 1.6.99.3) activity declined in liver and intestine, but not in skin or brain. 7. The activation coefficient of glutathione reductase was correlated strongly with nearly all the riboflavin-sensitive variables measured, once equilibrium had been reached in this chronic deficiency model, and it was particularly strongly correlated with hepatic and renal FAD levels. Under equilibrium conditions, therefore, it appears to represent a good index of the extent of riboflavin deficiency, and significant changes in flavin levels and enzymes in the internal organs were detected even under conditions of marginal deficiency, associated with relatively small increases in the activation coefficient. PMID- 7470439 TI - Energy intakes on sledging expeditions. AB - 1. Previous measurements of energy intake on sledging journeys in Antarctica have given a mean intake of 14.2 MJ (Acheson, 1974; Campbell, 1975), markedly lower than values reported earlier (see Edholm & Goldsmith, 1966). The technique used (individual weighed-diet survey) was more detailed and could be assumed to be more accurate than most of the earlier work where intakes had been largely inferred from the known energy content of food boxes. In the present study an individual weighed-diet survey was carried out on male subjects during a summer manhauling journey on the east coast of Greenland. 2. Mean daily energy intake of six subjects over 33 d travelling was 16.5 MJ. Mean weight loss was 2.3 kg, probably accounted for entirely by fat loss. Weight loss occurred despite the presence of excess food. Mean daily energy intake rose gradually but persistently over the 5 weeks of the journey despite a constant level of activity and to 20.1 MJ during the 4 d rest at the end of the journey. 3. Intakes were thus higher than those found in the earlier Antarctic studies (Acheson, 1974; Campbell, 1975) but not as high as intakes reported previously (see Edholm & Goldsmith, 1966). The fact that weight loss occurred despite the presence of excess food was ascribed to the monotonous nature of the diet. The fact that energy intake rose persistently over the 5-week study period may imply that a new state of balance of intake and expenditure was reached. PMID- 7470440 TI - Energy requirements of antarctic sledge dogs. AB - 1. Eighteen male sledge dogs were weighed immediately on removal from their sheltered winter quarters at Halley Bay, Antarctica (75 degrees 31' s, 26 degrees 42' W) and weekly thereafter for 14 weeks. The first 2 weeks they were tethered and inactive and the following 12 weeks travelled an average of 10.9 km/d fully laden. Daily energy intake during winter and while tethered averaged 18-25 MJ/d and while travelling 13.9 MJ/d. 2. Mean weight loss during the 2 weeks of inactivity was 2.3 kg despite an energy intake almost twice the recommended requirement. During the 12 weeks travelling energy intake decreased to 13.9 MJ/d but the weight loss stopped. Weather conditions at this time were becoming progressively milder. 3. The evidence suggests that sledge dogs are capable of high levels of cold-induced and possibly diet-induced thermogenesis and that these factors, particularly the former should be taken into account when designing dog rations and feeding schedules. PMID- 7470441 TI - A study of the effects of dietary added cupric oxide on the laying, domestic fowl and a comparison with the effects of hydrated copper sulphate. AB - 1. An experiment is reported in which copper, as cupric oxide, was fed to two breeds of laying hen for 336 d at levels equivalent to 150, 300, 450, 600 and 750 mg added Cu/kg diet. The results obtained were compared with those found using similar diets to which the Cu was added as CuSO4.5H2O. 2. Addition of the CuO had no effect on food intake, food conversion efficiency, body-weight or egg production. The CuSO4 addition caused the quadratic response of food intake and the adverse effects on food intake, egg production and body-weight noted in previous experiments. 3. The CuO had no effect on liver, kidney, ovary, oviduct or gizzard weight per unit body-weight while the CuSO4 decreased these with the exception of gizzard weight which was significantly increased. 4. CuO addition did not affect liver Cu concentration but CuSO4 caused a substantial increase of liver Cu especially at the 750 mg Cu/kg level of addition. 5. CuO addition had no effect on liver lipid concentration but this was depressed at the highest level of CuSO4 addition. Effects on individual fatty acids are presented but no specific conclusions have been reached. PMID- 7470443 TI - Degradation of deoxyribonucleic acid by a 1,10-phenanthroline-copper complex: the role of hydroxyl radicals. AB - Degradation of deoxyribonucleic acid (DNA) by 1,10-phenanthroline has been shown to require Cu(II), a reducing agent, and O2. Other metal ions do not substitute for Cu(II), and degradation of DNA is inhibited by metal ions that can form stable complexes with 1,10-phenanthroline, such as Co(II), Cd(II), Ni(II), or Zn(II), as well as by chelators that can bind copper, such as triethyltetraamine, neocuproine, or ethylenediaminetetraacetic acid (EDTA). Neocuproine, a specific copper chelator, is more effective than EDTA in inhibiting the breakdown of DNA. The degradation of DNA shows a requirement for a reducing agent which can be satisfied by either ascorbate or a thiol. A free radical generating system, e.g., xanthine oxidase-hypoxanthine, can substitute for the reducing agent. DNA degradation, in the presence of either an organic reducing agent or xanthine oxidase-hypoxanthine, is inhibited by hydroxyl radical scavengers and by catalase, suggesting that hydroxyl radical is the reactive species in DNA degradation and that hydrogen peroxide is an intermediate in hydroxyl radical generation. PMID- 7470442 TI - Initiation of reovirus transcription by inosine 5'-triphosphate and properties of 7-methylinosine-capped, inosine-substituted messenger ribonucleic acids. AB - Inosine 5'-triphosphate (ITP) can be utilized in place of guanosine 5' triphosphate (GTP) for both the initiation and the elongation steps of reovirus transcription, resulting in the synthesis of mRNAs containing 5'-terminal m7IpppIm and internal pI. The apparent molecular weights of the I-substituted products were altered as a consequence of the absence of G-C base pairs and accompanying loss of ordered structure. The migration of I-substituted RNAs in agarose gels and glycerol gradients was similar to glyoxal-treated transcripts; i.e., it decreased 2-fold as compared to the corresponding untreated G-containing mRNAs. 7-Methylinosine-capped (m7I-capped), I-substituted transcripts readily attached to wheat germ 80S ribosomes. Unlike native G-containing mRNAs, they also formed heavier complexes that sedimented faster than 80S complexes even in the presence of the nonhydrolyzable ATP analogue AMPP(NH)P and elongation inhibitor sparsomycin. I-substituted molecules that were capped posttranscriptionally to form m7G-capped 5' ends yielded mostly 80S monosomes, consistent with a strong influence of 5'-terminal structure on initiation of translation. Under limited conditions of initiation, I-substituted RNAs outcompeted G-containing transcripts for ribosome attachment. Although the results are consistent with enhanced binding and freer movement of ribosomes on unstructured templates, synthesis of acid-precipitable polypeptides in wheat germ extract directed by I-substituted RNAs was 15-fold less than with G-containing mRNAs. PMID- 7470444 TI - Interaction of aluminum species with deoxyribonucleic acid. AB - Interactions of aluminum with deoxyribonucleic acid (DNA) have been studied by thermal denaturation, circular dichroism, and fluorescent dye binding; a pH- and concentration-dependent alteration in the interaction of aluminum with DNA was observed. Three distinguishable complexes are produced when DNA is denaturated at pH 5.0-7.5 and in aluminum to DNA mole ratios of 0-0.7. Complex I appears at neutral pH and stabilizes a portion of DNA. Complex II is observed at acidic pH, destabilizes a fraction of the DNA double-helical molecule, and produces intrastrand cross-links. Complex III occurs at all pHs, is maximal at intermediate pH values, and is characterized by a noncooperative melting profile and cross-linking at low pH (less than 6.0). The DNA in complexes II and III can be renatured by treatment with either ethylenediaminetetraacetic acid (EDTA) or a high concentration of sodium chloride. The properties of complexes I and II are consistent with what could be expected for DNA complexes of Al(OH)2+ and Al3+, respectively. Complex III has intermediate properties that are consistent with a structure in which both ions bind the DNA simultaneously. The characteristics of complex III depend on the ratio of Al3+/Al(OH)2+ in solution. Aluminum-DNA complexes differ from other metal-DNA complexes in that melting profiles under many conditions are biphasic. Apparently more than one form of DNA can exist at any time in the presence of aluminum. The different DNA-aluminum complexes, which arise from the multiple species of aluminum in aqueous solution, lead to a variety of reactions with DNA. PMID- 7470445 TI - Studies on the mechanism of membrane fusion: kinetics of calcium ion induced fusion of phosphatidylserine vesicles followed by a new assay for mixing of aqueous vesicle contents. AB - We describe an assay for following the mixing of aqueous contents during fusion of phospholipid vesicles. Terbium is encapsulated as the Tb(citrate)3(6-) chelation complex in one population of vesicles, dipicolinic acid (DPA) in another. Vesicle fusion results in the formation of the fluorescent Tb(DPA)3(3-) chelation complex. The presence of EDTA (0.1 mM) and Ca2+ (greater than 1 mM) prevents the formation of the Tb/DPA complex in the external medium. We have studied the Ca2+-induced fusion of small or large unilamellar vesicles (SUV or LUV, respectively) composed of phosphatidylserine (PS). In addition, vesicle aggregation was monitored by light scattering, and release of vesicle contents was followed by carboxyfluorescein (CF) fluorescence enhancement. The addition of Ca2+ induced an immediate enhancement in Tb fluorescence with both SUV and LUV, which occurs on the same time scale as aggregation but much faster than the release of CF. The release of contents from LUV occurs with a considerable delay. It is estimated that the initial fusion of SUV is accompanied by 10% leakage of the internal volume per fusion event; in contrast, fusion of LUV is essentially nonleaky. Massive release of vesicle contents appears to be a secondary phenomenon related to the collapse of fused vesicles. The initial rate and the extent of Tb fluorescence enhancement are markedly dependent on the Ca2+ concentration. Threshold Ca2+ concentrations are 1.2 and 2.4 mM for SUV nd LUV, respectively. At saturating Ca2+ concentrations (greater than 10 mM), the rate of fusion of LUV is slightly lower than that of SUV at the same vesicle concentration. At any Ca2+ concentration, the rates of both SUV and LUV fusion are consistent with vesicle aggregation being rate limiting. When measured at a subsaturating Ca2+ concentration, fusion is essentially second order over a wide range of relatively low vesicle concentrations, whereas at higher vesicle concentrations the order is decreased. This suggests that at high vesicle concentrations (and at relatively low Ca2+ concentrations) aggregation may proceed faster than fusion. PMID- 7470446 TI - Studies on the mechanism of membrane fusion: role of phosphate in promoting calcium ion induced fusion of phospholipid vesicles. AB - The role of phosphate in enhancing the calcium-induced fusion of phosphatidylserine (PS) vesicles has been examined by using the new fluorescent (terbium/dipicolinic acid) assay described by Wilschut et al. (1980) [Wilschut, J., Duzgunes, N., Fraley, R., & Papahajopoulos, D. (1980) Biochemistry (preceding paper in this issue)]. In the presence of physiological levels of phosphate, the calcium concentration required for fusion of PS vesicles was lowered significantly (3-4-fold), and the rate of vesicle fusion was increased dramatically (up to 1000-fold). The fusion of PS vesicles by calcium and phosphate is shown to be specific and critically dependent on temperature, pH, ion concentrations, and the composition of the calcium phosphate crystalline phase present during the incubation. The results indicate that a significant enhancement in vesicle fusion occurs only when calcium phosphate precipitation is initiated in the presence of PS vesicles, suggesting that crystal nucleation on the vesicle surface is a prerequisite for fusion. Calcium and phosphate were shown to promote phospholipid phase separations and vesicle fusion under conditions (e.g., mixtures of PS and phosphatidylcholine) in which calcium alone is ineffective, indicating that formation of PS calcium phosphate complexes may facilitate the molecular segregation of PS into distinct domains. These experiments underline the important role that phosphate may play in calcium mediated fusion phenomena in biological membranes. PMID- 7470447 TI - Mass action kinetics of phosphatidylserine vesicle fusion as monitored by coalescence of internal vesicle volumes. AB - The kinetics of Ca2+-induced fusion of sonicated phosphatidylserine vesicles is analyzed by means of the mass action model. The results of calculations are shown to simulate the experimental results for the mixing of aqueous vesicle volumes, release of vesicle contents and for the observed increase in light scattering [Wilschut, J., Duzgunes, N., Fraley, R., & Papahadjopoulos, D. (1980) Biochemistry (first of three papers in this issue)]. The calculations give the distribution of vesicle sizes during the initial stages of the fusion process and an estimate for the occurrence of multiple fusion events. It is estimated that during the first few seconds from the beginning of the fusion process in the above systems only a small fraction of the material trapped will leak during each fusion event. The fraction of material which leaks per fusion event is further reduced with increased Ca2+ concentrations. The values of the rates of fusion which describe the above experiments suggest that the rate limiting step of the overall fusion reaction is the aggregation and close approach of vesicles to each other rather than the fusion event per se. PMID- 7470448 TI - Raman spectroscopic study of the interactions of dimyristoyl- and 1-palmitoyl-2 oleoylphosphatidylcholine liposomes with myelin proteolipid apoprotein. PMID- 7470450 TI - Effects of adenosine 5'-diphosphate on bovine glutamate dehydrogenase: diethyl pyrocarbonate modification. AB - Initial rate kinetic studies and reduced coenzyme binding studies with bovine glutamate dehydrogenase have shown that an enzyme-NAD(P)H-glutamate abortive complex is a major participant in the overall enzyme-catalyzed reaction. The allosteric regulator ADP is shown to activate the enzyme by destabilizing this abortive complex. Destabilization of this abortive complex with concomitant activation is also achieved by the ethoxyformylation of a single histidine residue per subunit in the hexameric enzyme. ADP, in addition to its activatory effects, is shown to (a) remove the nonlinearity from the Lineweaver-Burk plots which is attributable to negative cooperativity and (b) inhibit the enzyme as a competitive inhibitor with respect to coenzyme under the appropriate conditions. Ethoxyformylation with diethyl pyrocarbonate, which mimics the activatory effects of ADP, has no effect on the negative cooperativity shown by this enzyme. A model for the action of ADP is proposed in which ADP activates glutamate dehydrogenase by binding to a regulatory binding site and blocks the negative cooperativity by mimicking the natural coenzyme at the active site of the enzyme. PMID- 7470449 TI - Comparative lateral diffusion of fluorescent lipid analogues in phospholipid multibilayers. PMID- 7470451 TI - pH kinetic studies of bovine brain hexokinase. AB - The variation of kinetic parameters with pH was examined for bovine brain hexokinase with glucose and MgATP as substrates. The -log V1 and -log (V1/Km) profiles for both substrates were examined and seen to decrease below pH 6.5. All profiles asymptotically approached slopes of -1, indicating that the loss of activity in each instance was due to the protonation of a single group on the enzyme. Analysis of the data indicated two ionizable groups were involved in the reaction. One functions in the binding of ATP and in catalysis while the other participates in the binding of glucose. The -log V1 profiles both showed a "hump" attributed to a loss of activity in the pH region 7.5-5.5. Addition of aluminum ions to the reaction mixture increased the magnitude of the hump, but the inhibition was abolished by the addition of citrate. Kinetic studies carried out at pH 7 indicated that aluminum was a competitive inhibitor with respect to ATP and noncompetitive with respect to glucose. However, secondary plots of the kinetic data were nonlinear, concave downward, indicating that the inhibition is not of a simple type. Possible explanations for this phenomenon are presented. PMID- 7470452 TI - Resonance energy transfer between catalytic sites of bovine liver uridine diphosphoglucose dehydrogenase. PMID- 7470453 TI - Polysome-dependent in vitro translation system capable of peptide chain reinitiation. AB - A sensitive in vitro translation system has been developed which makes use of cellular polysomes as the source of mRNA and ribosomes. The soluble factors are derived from the preincubated S-30 fraction by centrifugation through a discontinuous sucrose gradient. Of the four fractions tested, fraction 1 (topmost fraction in the gradient) and fraction 2 (fraction sedimenting in 0.5 M sucrose) were stimulatory. These two fractions together yield the highest activity, corresponding to about 125 times the background incorporation. The polysome directed system exhibits optimal activity in the range 1.8-2 mM Mg2+ and 125-175 mM KCl. The polysome-directed in vitro products exhibit a complexity comparable to the in vivo products resolved on the two-dimensional polyacrylamide gels of O'Farrell [O'Farrell, P. (1975) J. Biol. Chem. 250, 4007-4021]. The system is capable of active chain reinitiation as indicated by partial inhibition by 7 methylguanosine 5'-monophosphate and pactomycin and N-terminal end analysis of in vitro products. This system can also translate polysomes from diverse tissues such as mouse liver, rat liver, and rat brain. The levels and also the authenticity of translation of rat liver albumin and mouse liver carbamoyl phosphate synthetase I were tested by immunoprecipitation with monospecific antibodies. The results show that the major as well as the minor translation products are synthesized in this system at levels comparable to the physiological levels. PMID- 7470454 TI - A major intermolecular cross-linking site in bovine dentine collagen involving the alpha 2 chain and stabilizing the 4D overlap. AB - Approximately 20% of the radioactivity incorporated into the dentine collagen of unerupted bovine molars after reduction with tritiated sodium borohydride was recovered in a cyanogen bromide peptide fraction of Mr 61 000 following chromatography on agarose A5m. After rechromatography on agarose A1.5m, this fraction was resolved into ten components by gel isoelectric focusing. Of these components, nine (the most acidic) were tritiated and contained the reduced cross links dihydroxylysinonorleucine and hydroxylysinonorleucine. The amino acid compositions were consistent with the identification of each of these components as alpha 2CB3,5 linked to one or two small peptides. By limited Edman degradation, with and without prior digestion with pyroglutamate aminopeptidase (EC 3.4.11.8), these small peptides were identified as alpha 1CB0,1 and alpha 2CB1, occurring in a ratio of approximately 2:1. Specific cleavage with cathepsin D revealed that all the cross-link was associated with the C-terminal one-third of the alpha 2 chain, thus fixing the displacement of the participating molecules at 4D. The content of the known reducible cross-links present in these peptides, calculated from the specific activity of the reductant, was sufficient to account for only 10--20% of the cross-linking actually found, suggesting that stabilization is mainly through nonreducible cross-links of as yet undetermined structure. By quantitative analysis of homoserine content and semiquantitative amino-terminal analyses, it was determined that virtually all of the alpha 2 chain of bovine dentine collagen is cross-linked in this manner. One cross-link per molecule in this location could made a major contribution to the mechanical stability of the insoluble collagen fibrils in this tissue. PMID- 7470455 TI - Conformation of phosphatidylethanolamine in the gel phase as seen by neutron diffraction. AB - For confirmation of some general aspects of phospholipid conformation in membranes and extension of previous neutron diffraction studies on dipalmitoyllecithin, measurements have now been made on 1,2-dipalmitoyl-sn glycero-3-phosphoethanolamine (DPPE) in the gel phase by the same method. Three selectively deuterated samples were investigated; in one of the specimens the first methylene segment close to the glycerol backbone in both chains was deuterated, and in the other two samples one of the methylene segments in the phosphoethanolamine group was replaced by CD2. Together with the undeuterated DPPE, these probes were investigated at very low water content (about 1.5--2 molecules of water per lipid) as oriented samples at 25 degrees C. The intensities of the first 12 reflections were collected and phased, and the mean positions of the segments were determined. The results confirm the idea that the conformation of a DPPE molecule in the gel state is very similar to the crystal structure of rac-1,2-dilauroyl-sn-glycero-3-phosphoethanolamine. The two main features are (1) the chains remain in all all-trans conformation having an axial displacement of about 3--4 A, (2) the zwitterionic dipoles in the head groups of both compounds are found to be aligned almost parallel to the bilayer surface. The main advantage of the method results in the fact that the combination of neutron scattering with selectively deuterated probes allows the determination of the mean label position to an accuracy of up to +/- 1 A. PMID- 7470456 TI - Identification of N-acylethanolamine phosphoglycerides and acylphosphatidylglycerol as the phospholipids which disappear as Dictyostelium discoideum cells aggregate. AB - The cellular slime mold Dictyostelium discoideum contains a phospholipid fraction which comprises 10% of the phospholipids in the early developing amoebae and disappears during the aggregation stage of development. As a first step in studying its metabolism, the composition of the fraction has been determined. It was easily isolated by preparative silicic acid thin-layer chromatography because its Rf was considerably higher than most commonly encountered phospholipids. Its Rf was the same as synthetic phosphatidyl-N-acylethanolamine and synthetic acylphosphatidylglycerol (also called semilysobisphosphatidic acid). Strong absorption peaks characteristic of amide bonds in the infrared spectrum of the isolated D. discoideum phospholipid showed that N-acylethanolamine phosphoglycerides were present. The presence of acylphosphatidylglycerol was revealed when mild alkaline hydrolysis of the lipid fraction produced glycerophosphorylglycerol as the only water-soluble, phosphate-containing product. The composition of the fraction was determined by chemical analysis and thin-layer chromatography of the intact phospholipids and their partially or completely hydrolyzed products. The composition of the fraction was 30% diacylglycerophosphoryl-N-acylethanolamine, 50% alkenylacylglycerophosphoryl-N acylethanolamine, and 20% acylphosphatidylglycerol. The stereoconfiguration of the glycerophosphorylglycerol moiety of the acylphosphatidylglycerol was found to be sn-3-glycerophosphoryl-sn-1'-glycerol. PMID- 7470457 TI - Primary structure of murine major histocompatibility alloantigens: amino acid sequence of the cyanogen bromide fragment Ia (positions 139-228) from the H-2Kb molecule. AB - The complete amino acid sequence of the cyanogen bromide (CNBr) fragment Ia (CN Ia) from the murine histocompatibility antigen H-2Kb has been obtained by using radiosequence methodology. This glycopeptide is the largest CNBr cleavage product of the H-2K molecule and extends from position 139 to position 228. The sequence determined for CN-Ia was Ala-Ala-Leu-Ile-Thr-Lys-His-Lys-Trp-Glu-Gln-Ala-Gly-Glu Ala-Glu-Arg-Leu-Arg-Ala -Tyr-Leu-Glu-Gly-Thr-Cys-Val-Glu-Trp-Leu-Arg-Arg-Tyr-Leu Lys-Ans-Gly-(Asn)-Ala- Thr-Leu-Leu-Arg-Thr-Asp-Ser-Pro-Lys-Ala-His-Val-Thr-His His-Ser-Arg-Pro-Asp-Asp -Lys-Val-Thr-Leu-Arg-Cys-Trp-Ala-Leu-Gly-Phe-Tyr-Pro-Ala Asp-Ile-Thr-Leu-Thr-Tr p-Gln-Leu-Asn-Gly-Glu-Glu-Leu-Ile-Gln-Aps-Met. The data were obtained by analysis of fragments derived by thrombic, tryptic, chymotryptic, and V8 protease digestion of CN-Ia. A carbohydrate moiety is attached to Asn at position 176. Homology between this 90 amino acid stretch of H 2Kb and HLA-B7 [Orr, H. T., Lopez de Castro, J. A., Lancer, D., & Strominger, J. L. (1979) Biochemistry 18, 5711] is 68%, and differences are noted at positions 176, 177, and 178 which in the H-2 molecule are the attachment region for a second carbohydrate moiety. No carbohydrate was detected in this position for HLA B7 [Orr, H. T. Lopez de Castro, J. A. Lancet, D., & Strominger, J. L. (1979) Biochemistry 18, 5711]. PMID- 7470458 TI - Primary structure of murine major histocompatibility complex alloantigens: completion of the sequence of the amino-terminal 284 residues of H-2Kb. AB - The primary structure of the COOH-terminal cyanogen bromide (CNBr) cleavage fragment Ic (CN-Ic) of the extracellular portion of the murine histocompatibility antigen H-2Kb has been completed. CN-Ic contains a site of papain cleavage which has been utilized for solubilizing H-2Kb by cleaving off the membrane integrating portion of the molecule. The amino acid sequence of CN-Ic has been determined by using peptides recovered after trypsin digestion of CN-Ic before and after blockage of lysine groups with citraconic anhydride. Overlapping sequences for the tryptic fragments were obtained by amino-terminal sequence analysis. The sequence of fragment CN-Ic, which spans residues 229-284 in H-2Kb, is as follows: Glu-Leu-Val-Glu-Thr-Arg-Pro-Ala-Gly-Asp-Gly-Thr-Phe-Gln-Lys-Trp-Ala-Ser-Val-Val Pro-Leu-Gly-Lys-Glu-Gln-Tyr-Tyr-Thr-Cys-His-Val-Tyr-Gln-Gln-Gly-Leu-Pro-Gln-Pro Leu-Thr-Leu-Arg-Trp-Asp-Glu-Pro-Pro-Ser-Thr-Val-Ser-Asn-Met. This amino acid sequence determination completes the primary structure of the amino terminal 284 residues of H-2Kb, that portion of this histocompatibility antigen which is external to the cell membrane and which contains antigenic determinants. It was also possible to identify Val-281 as a papain cleavage site within CN-Ic. The completed structure was analyzed solely by radiochemical methods. The structure obtained for H-2Kb is 71% homologous to the reported structure of HLA-B7, a human homologue. PMID- 7470459 TI - Synthetic cation transport peptides: calcium transport across phospholipid membranes. AB - Molecular aspects of peptide-mediated calcium transport are examined through the study of the cation transport properties of a series of synthetic cyclic octapeptides. These peptides, of general structure cyclo[Glu(OR1)-Sar-Gly-(N R2)Gly]2 (R1 = H or benzyl ester; R2 = cyclohexyl, n-hexyl, or n-decyl) (and an Asp analogue), contain central binding cavities of geometry and dimensions similar to calcium-binding sites in proteins. Transport in Pressman cells ("thick liquid membranes") demonstrated the ionophorous activity of the synthetic peptides; among physiologically abundant cations, the order of selectivity was Ca2+ greater than Na+, K+ much greater than Mg2+. Cation competition studies further showed that cyclo[Glu(OBz)-Sar-Gly-(N-cyclohexyl)Gly]2 (CYCLEX-2E) is essentially a calcium-specific transport peptide whenever calcium is present. When the CYCLEX-2E peptide was added to a suspension of 45Ca2+-loaded sonicated phosphatidylcholine (PC) vesicles in a dialysis sac, the vesicles were completely emptied of internal calcium. Controls using [14C]sucrose established that CYCLEX 2E caused no nonspecific membrane damage. Calcium efflux experiments using several salts of calcium (including 36C1-, [14C]acetate, [14C]succinate, and 35SO4(2-)) suggested that these anions do not specifically accompany the Ca2+ peptide active transporting species across the phospholipid membrane. However, when 45Ca2+-loaded PC vesicles were suspended in mental-free buffer and treated with CYCLEX-2E peptide, calcium efflux did not occur until calcium or sodium chloride was added to the external medium. PMID- 7470460 TI - Selectivity of fluorescent lipid analogues for lipid domains. AB - We have examined the phase partition preferences of the even chain length (n = 10 22) diacyl-3'3'-indocarbo-cyanine iodides (Cn diI) incorporated in disaturated lecithin (PC) vesicles. Two parameters were used to determine this phase preference: (i) the direction of shift of the phase transition temperature (Tm) induced by the dyes and (ii) the self-quenching of fluorescence due to aggregation in the gel phase of those dyes which preferentially partition into the fluid. Dyes that lower Tm preferentially partition into the fluid phase; those that raise Tm preferentially partition into the gel. By these criteria in dimyristoyl-PC, C10 diI and C12 diI preferentially partition into the fluid phase, C14 diI and C16 diI show no preferential partition, C18 diI preferentially partitions into the gel, and C20 diI and C22 diI preferentially partition into the fluid. In dipalmitoyl-PC, the pattern of preference is identical with that observed in dimyristoyl-PC, only shifted to longer chain length diI's by two carbons. Diffusion measurements by fluorescence photobleaching recovery of these dyes in gel-phase multilayers showed tham all to be immobile, D less than 10(-10) cm2/s, while in fluid-phase multilayers thay all had diffusion coefficients of D approximately 10(-8) cm2/s independent of chain length. In mixed-phase multilayers, however, each Cn diI showed mobile fraction which reflected its phase-partition preference. PMID- 7470461 TI - Effects of nocodazole on structures of calf brain tubulin. AB - The interaction of nocodazole with calf brain tubulin was studied to determine the effect of such interaction on the structure of tubulin. The effect of nocodazole on the self-association of tubulin was monitored by turbidity measurements and velocity sedimentation. Sedimentation patterns indicate that nocodazole neither induces tubulin to undergo self-association to form higher orders of aggregate nor does it perturb the equilibrium of the reaction leading to the formation of 42S double-ring structures although nocodazole binds to both the tubulin dimers and the polymeric form. Nocodazole does, however, inhibit the in vitro reconstitution of microtubules, and the presence of microtubule associated proteins does not amplify the inhibitory effect of the drug. The conformational changes in tubulin upon binding of nocodazole were monitored by differential spectroscopy, circular dichroism, fluorescence, and chemical modification of sulfhydryl residues. Results from these studies show that the sulfhydryl residues become more accessible to chemical modification. In contrast, the binding of nocodazole does not significantly alter the net environment of tryptophan chromophores. These residues are apparently not all located on the surface of the tubulin molecule and at least some are partially buried. PMID- 7470462 TI - Dissociation of antithrombin III--thrombin complex. Formation of active and inactive antithrombin III. AB - Dissociation of the complexes formed during the reaction of thrombin with antithrombin III was studied by determining the kinetics of release of thrombin and active antithrombin III from the complexes. The rate of thrombin release increased gradually between pH 7.0 and 8.5 and then sharply increased at pH 9.0. The rate of active antithrombin III release did not change significantly over the same pH range. The results suggested that thrombin is released after reaction with antithrombin III via two pathways which are differentially affected by the pH of the solution. At pH 7.0 thrombin is released predominantly by dissociation of a non-acylated enzyme--inhibitor complex since active antithrombin III was released in relatively low amounts. The results suggested that thrombin inhibition by antithrombin III does not require the formation of an acyl bond between the active-site serine of thrombin and a carbonyl carbon of antithrombin III. The release of thrombin by acylation--deacylation appeared to be limited by the rate of enzyme acylation which was favored by more alkaline pH. This further suggests that acyl bond formation is a secondary reaction that can occur during thrombin inhibition by antithrombin III but is not necessary for thrombin inhibition. PMID- 7470463 TI - Adenosine analogues as substrates and inhibitors of S-adenosylhomocysteine hydrolase. AB - In the reaction adenosine + L-homocysteine = S-adenosyl-L-homocysteine, catalyzed by S-adenosylhomo-cysteine hydrolase from beef liver (EC 3.3.1.1), 11 nucleosides are able to substitute for adenosine to generate their corresponding S nucleosidylhomocysteine congeners: 3-deaza-adenosine, 2-aza-3-deazaadenosine, nebularine (purine ribonucleoside), formycin, N6-methyladenosine, 8-azaadenosine, adenosine N1-oxide, pyrazomycin, 8-aminoadenosine, inosine, and the carbocyclic analogue of adenosine [(+/-)-aristeromycin]. S-Adenosylhomocysteine hydrolase from lupin seeds is able to utilize all of these nucleosides except inosine to synthesize analogues of S-adenosylhomocysteine. There is no correlation between the ability of these nucleotides to function as substrates and their inhibitory potencies, except in the case of 3-deazaadenosine. The carbocyclic analogue of adenosine is the most potent inhibitor of S-adenosylhomocysteine hydrolase with a Ki of 5 X 10(-9) M. When incubated with 3T3-L1 fibroblasts, the carbocyclic analogue of adenosine caused a 20-fold increase in the cellular concentration of S-adenosyl-homocysteine. The cellular generation of S-2-aza-3-deaza adenosylhomocysteine was observed when 3T3-L1 fibroblasts were incubated with 2 aza-3-deazaadenosine. PMID- 7470464 TI - Effects of self-association of ornithine aminotransferase on its physicochemical characteristics. AB - Previous work in this laboratory [e.g., Peraino, C., Bunville, L. G., & Tahmisian, T. N. (1969) J. Biol. Chem. 244, 2241--2249, and Morris, J. E., Peraino, C., & Strayer, D. (1974) Proc. Soc. Exp. Biol. Med. 147, 707--709] has shown that the molecular weight of ornithine aminotransferase (OAT) is concentration dependent. In the present study this property of OAT was further characterized by using sedimentation equilibrium centrifugation to determine the molecular weight of OAT in a range of enzyme concentrations. It was shown that OAT aggregates in a two-stage process as its concentration increases. The first stage involves the association of enzymatically active monomers into trimers, with association of the trimers into higher order aggregates occurring in the second stage. Decreasing the pH or raising the ionic strength enhances aggregation while raising the pH inhibits aggregation; however, the two-stage nature of the aggregation process was not affected by changes in pH and ionic strength. Kinetic analyses of purified enzyme showed that aggregation results in an increase in the kM for both substrates with the Vmax remaining constant, indicating that aggregation of monomers sterically hinders substrate binding. Increased Km values were also obtained for OAT sequestered in mitochondria from rats fed a high-protein diet to increase mitochondrial OAT levels. The higher Km values suggest that the elevation of OAT in vivo is accompanied by aggregation of the enzyme within the mitochondrion. We propose that the aggregation-dependent increase of Km in vivo has adaptive value in that it spares ornithine for use in the urea cycle. PMID- 7470465 TI - Protein-catalyzed phospholipid exchange between gel and liquid-crystalline phospholipid vesicles. AB - Bovine liver phospholipid exchange protein catalyzes the transfer of phosphatidylcholine between two populations of single bilayer phospholipid vesicles. Donor vesicles are prepared from egg phosphatidylcholine--phosphatidic acid--lactosylceramide (90:2:8) mol %); acceptor vesicles are prepared from phosphatidylcholine--phosphatidic acid (98:2 mol %). Activity is determined from the rate of transfer of 3H-labeled egg phosphatidylcholine from donor to acceptor vesicles in the presence of phospholipid exchange protein. Donor vesicles are quantitatively precipitated by Ricinus communis agglutinin, while acceptor vesicles remain in the supernate. When egg phosphatidylcholine acceptor vesicles over the temperature range 11--45 degrees C are used, a linear Arrhenius plot is obtained, in keeping with the observation that these membranes exist only in the liquid-crystalline state. When dimyristoylphosphatidylcholine acceptor vesicles under the same conditions are used, however, a biphasic plot is seen with decreasing transfer activity at lower temperatures. The discontinuity occurs at 31 degrees C and corresponds with the onset of the liquid-crystalline to gel phase transition. The incorporation of cholesterol into dimyristoylphosphatidylcholine vesicles at a concentration sufficient to abolish the thermotropic phase transition yields a monophasic Arrhenius plot of transfer activity. The results indicate that bovine liver phospholipid exchange protein interacts catalytically with phospholipid bilayer vesicles composed of saturated or unsaturated phosphatidylcholines but preferentially with liquid-crystalline membranes. PMID- 7470466 TI - Calorimetric evaluation of the existence of separate domains in bovine prothrombin. PMID- 7470467 TI - Interaction of succinate--ubiquinone reductase (complex II) with (arylazido)phospholipids. AB - The interaction of purified succinate dehydrogenase and succinate--ubiquinone reductase (complex II) with lipids was explored by using two (arylazido)phospholipids, one with the reactive nitrene in the head-group region of the bilayer [1-palmitoyl-2-(2-azido-4-nitrobenzoyl)-sn-glycero-3 [3H]phosphocholine (PLII)] and one with the nitrene on the methyl terminus of one of the fatty acid chains [1-myristoyl-2-[12-[(2-azido-4 nitrophenyl)amino]lauroyl]-sn-glycero-3-[14C]phosphocholine (PLI)]. Protein was reacted with vesicles of egg lecithin containing radioactive (arylazido) phospholipids and the covalent cross-linking of lipid and protein induced by irradiation under UV light. Purified succinate dehydrogenase was found to bind to lipid vesicles through both subunits as both were labeled by PLII. The smaller subunit was inserted into the interior of the bilayer and labeled by PLI. Complex II was found to interact with lipid vesicles, with the smaller subunit of succinate dehydrogenase, CII-3, and CII-4 all inserted into the interior of the bilayer. The large subunit of succinate dehydrogenase was found to be held above the bilayer in complex II and not labeled by either probe. Results are used to derive a picture of the arrangement of subunits in complex II and to evaluate the utility of (arylazido)-phospholipids in membrane studies. PMID- 7470468 TI - Differential scanning calorimetry of chloroplast membranes: identification of an endothermic transition associated with the water-splitting complex of photosystem II. AB - The structure of spinach thylakoid membranes has been investigated by sensitive differential scanning calorimetry. Six endotherms are observed between 20 and 85 degrees C, corresponding to order--disorder transitions of different structural domains within the thylakoid membrane. In a medium of relatively high ionic strength, endothermic transitions occur at 42, 54, 65, 72, 79, and 84 degrees C, with the 65 degrees C transition being particularly prominent. At a lower ionic strength, transitions are centered at 44, 61, 66, 70, 78, and 83 degrees C. The 42--44 degrees C endothermic transition (the A transition) can be correlated with the modification of three electron-transport components or properties associated with photosystem II: (i) release of manganese from the membrane, (ii) the loss of O2 evolution with water as a donor, and (iii) a decrease in the redox potential of the hydroquinone-reducible cytochrome b-559. Both the A transition and the ability to evolve O2 are irreversibly lost after heating to 49 degrees C and also after exposure to trypsin, suggesting the involvement of protein in this transition. The interpretation of these observations is that one effect of the A transition involves the thermal disruption of a protein component on the donor side of photosystem II. PMID- 7470469 TI - Isolation and characterization of neurofilaments from mammalian brain. PMID- 7470470 TI - A MAP-2-stimulated protein kinase activity associated with neurofilaments. PMID- 7470471 TI - Photoaffinity labeling of Crotalus atrox phospholipase A2 by a substrate analogue. AB - A photolabile analogue of phosphatidylethanolamine (photolabile PE analogue), 1,2 di-O-hexylglycero-3-(ethyl diazomalonamidoethyl phosphate), was synthesized in nonisotopic and 14C-radiolabeled form and in both the L configuration (that of the naturally occurring phospholipids) and the racemic form. When the unlabeled racemic compound was photolyzed in the presence of phospholipase A2 of Crotalus atrox, extensive enzyme inactivation was observed. The rate of inactivation was stimulated by Ca2+ and by formation of micelles of the photolabile compound. The dihexyl ether analogue of phosphatidylethanolamine protected the enzyme from inactivation. Phospholipase A2 gave rise to a covalently labeled polypeptide when irradiated in the presence of either L or racemic 14C-labeled photolabile PE analogue. The racemic compound labeled both the N-terminal region (residues 1- 15) and the central region (residues 43--97) of the polypeptide while the L compound labeled only the N-terminal region. The lone methionine at position 10 of the C. atrox phospholipase A2 permitted degradation by cyanogen bromide, which showed that labeling by the L compound was restricted to the first ten amino acid residues at the N-terminal end. Phospholipase A2 has an absolute specificity for L-phospholipids, and D-phospholipids are competitive inhibitors. The results of these studies underscore the importance of the head-group region of the phospholipid in phospholipase--substrate interactions and suggest that the two optical isomers of the substrate may be rather differently oriented on the enzyme surface. PMID- 7470472 TI - Dilatometry and calorimetry of saturated phosphatidylethanolamine dispersions. AB - The specific volumes of a series of saturated phosphatidylethanolamine dispersions with 12, 14, and 16 carbon atoms per chain have been measured in the region of the chain melting transition, Tm. The change in specific volume at Tm for the 12 and 14 carbon compounds are 0.0160 and 0.0204 mL/g, respectively. Comparisons are drawn between this class of lipids and phosphatidylcholines. In both cases, Tm extrapolates with increasing chain length to the melting point of polyethylene. Both types of lipids appear to be packed in a similar way below Tm. One major difference is that dilaurylphosphatidylethanolamine undergoes a second transition above Tm. PMID- 7470473 TI - Photochemistry and fluorescence of bacteriorhodopsin excited in its 280-nm absorption band. AB - Photochemical and fluorescence studies are carried out, exciting bacteriorhodopsin (BR) in its 280-nm absorption band. The data indicate that energy transfer takes place, with a quantum yield of 0.7--0.8, from excited tyrosines and tryptophans to the retinyl chromophore. All of the tyrosine and five to six tryptophan residues are completely quenched by the transfer process while one tryptophan is unquenched and one is partially (approximately 80%) quenched. Energy transfer to the chromophore leads to a photocycle identical with that triggered in (light adapted) bacteriorhodopsin by excitation within the visible absorption bands of the chromophore. The emissive properties of BR in the intact membrane are found equal to those of Triton X-100 solubilized BR monomer. The energy transfer data are discussed in terms of the available amino acid sequence and the electron density map of bacteriorhodopsin. Although such data cannot suggest a single fit between the sequence and the density map (one out of the 7! = 5040 possibilities), they do provide a criterion for testing any specific model for the structure of bacteriorhodopsin. PMID- 7470474 TI - Kinetic characteristics which distinguish two forms of calf thymus DNA polymerase alpha. AB - DNA polymerase alpha was isolated as previously described [Holmes, A. M., Hesslewood, I.P., & Johnston, I. R. (1974) Eur. J. Biochem. 43, 487]. This method yields five nuclease-free forms of alpha-polymerase, A1, A2, B, C, and D. Holmes and co-workers [Holmes, A. M., Hesslewood, I.P., Wickremasinghe, R. G., & Johnston, I.R. (1977) Biochem. Soc. Symp. 42, 17] have suggested that the C form is the core enzyme of alpha-polymerase and have demonstrated that removal of a protein subunit from the A1 form yields an enzyme with the physical properties of the C form. They did not investigate the function of the subunit because the A1 and C forms were not easily distinguished with biochemical kinetics. We have been able to demonstrate three kinetic differences between these forms: (1) the alpha A1-polymerase adds more nucleotides per binding event to activated DNA (is more processive) than does alpha-C-polymerase. (2) The synthetic activity of the alpha A1-polymerase is greater on a template with an average gap size of 65 nucleotides than it is on a template with an average gap size of 10 nucleotides whereas that of the alpha-C-polymerase is not. (3) The synthetic activity of the alpha-C polymerase is inhibited by high concentrations of activated calf thymus DNA (greater than 300 muM) whereas that of the alpha-A1-polymerase is not. The nature of the inhibitor was investigated and found to be a nuclear RNA component present in the DNA preparations. These kinetic differences may provide a means to assay for the protein subunit that converts alpha-C-polymerase to alpha-A1-polymerase, and provide a basis for isolation and characterization of other DNA replication association proteins. PMID- 7470475 TI - Molecular weight of undegraded plasma factor V. AB - The molecular size and immunochemical properties of the unfractionated factor V present in plasma collected by venipuncture into a broad-spectrum anticoagulant and platelet-inhibited mixture were compared with those reported for the isolated, single-chain factor V molecular of 330 000 daltons. The anticoagulant- plasma mixture included 0.28% trisodium citrate, 2 mM benzamidine hydrochloride, 0.02% soybean trypsin inhibitor, 2.0 mM diisopropyl phosphorofluoridate, 10 microM dansylarginine N-(3-ethyl-1,5-pentanediyl)amide, and 5 microM prostaglandin E1. The Stokes radius of unfractionated factor V present in highly inhibited plasma (93 A) is virtually identical with the Stokes radius predicted from the hydrodynamic data for the highly asymmetric, single-chain factor V molecule (91 A). With an expression which relates the Stokes radius and the sedimentation coefficient to the molecular weight of hydrodynamic units, the molecular weight obtained for factor V, using gel filtration data, is 336 000, in good agreement with the molecular weight determined from the sedimentation equilibrium, 330 000. In contrast, the Stokes radius for the factor Va present in serum is significantly smaller (50.5 A) and equivalent to the Stokes radius obtained upon activation of isolated factor V with thrombin. Immunochemical comparisons of the factor V present in the inhibited plasma and isolated factor V were conducted by using burro antibovine factor V antibody and the technique of immunoelectrophoresis. The factor V antigen present in both sources is immunochemically identical, as is the electrophoretic mobility of both factor V preparations. These data serve to justify the conclusion that the factor V isolated as a single-chain 330 000-dalton molecule corresponds to the factor V circulating in plasma. PMID- 7470476 TI - Estimation of globular protein secondary structure from circular dichroism. AB - A new method is developed in which a circular dichroism (CD) spectrum is analyzed directly as a linear combination of the CD spectra (from 190 to 240 nm) of 16 proteins whose secondary structures are known from X-ray crystallography. This avoids the dilemma encountered in previous methods of trying to define single reference CD spectra that were supposed to characterize such broad and variable classes as helix, beta sheet, beta turn, and "remainder". It also permits a more accurate and flexible analysis. The usual instability in using so many parameters is automatically controlled by a simple constrained statistical regularization procedure (similar to ridge regression). Sixteen tests were made by removing 1 spectrum at a time from the set of 16 and analyzing it in terms of the other 15. The product moment correlation coefficients between the computed fractions of helix, beta sheet, beta turn, and remainder and the fractions from the X-ray data were 0.96, 0.94, 0.31,, and 0.49, respectively. Thus, the helix and beta-sheet accuracy is very good. (The corresponding values calculated by a previous method with four reference spectra were 0.85, 0.25, --0.31, and 0.46.). PMID- 7470477 TI - Determination of protein--ligand equilibria by difference spectroscopy. Hemerythrin--ligand thermodynamic studies. AB - A difference spectrophotometric method for the rapid determination of equilibrium constants for protein--ligand interactions has been developed. The method requires no knowledge of the extinction coefficient of either reactants or products. Furthermore the method allows rapid determination of the temperature dependence of a reaction and thus leads to rapid determination of thermodynamic parameters. The method has been tested by following the interactions of ligands with hemerythrin, the nonheme iron, oxygen storage protein isolated from Phasocolopsis gouldii. The reactions were studied at various temperatures and ionic strengths, and standard thermodynamic parameters were determined. The standard thermodynamic parameters for the conversion of metaquohemerythrin to methydroxyhemerythrin were found to be delta H degrees = 5.8 +/- 1.3 kcal mol-1 and delta S degrees = -11.5 +/- 1.5 cal mol-1 deg-1. For the reaction of metaquohemerythrin with thiocyanate ion to produce metthiocyanatohemerythrin delta H degrees = --13.0 +/- 1.6 kcal mol-1 and delta S degrees - --25.3 +/- 5.5 cal mol-1 deg-1. For the reaction of thiocyanate ion with methydroxy-hemerythrin delta H degrees = --6.6 +/- 0.8 kcal mol-1 and delta S degrees = --38.3 +/- 4.0 cal mol-1 deg-1. Perchlorate ion decreases the affinity of metaquohemerhythrin for thiocyanate ion. This is reflected in both the entropy and enthalpy being more unfavorable for the reaction in the presence of perchlorate ion. PMID- 7470478 TI - Determination of a low pK for histidine-159 in the S-methylthio derivative of papain by proton nuclear magnetic resonance spectroscopy. AB - Proton NMR spectroscopy was used to study the ionization behavior of His-159 in a derivative of papain (papain-S-SCH3). In this catalytically inactive derivative of papain, the active-site thiol group of Cys-25 is S-methyl-thiolated so that it cannot form a thiolate anion. The pH dependence of the chemical shift of the C epsilon 1 H resonance of His-159 indicated a pK of 3.45 +/- 0.07 at 45 degrees C in 2H2O with no added ions other than those required for titration. In acetate buffers at an ionic strength of 0.05, the pK increased to 3.87 +/- 0.12. Conversion of papain-S-SCH3 to active papain at pH* 4.17 (at 45 degrees C and an ionic strength of 0.05) caused the position of the C epsilon 1 H resonance to change from a position indicative of partial protonation of His-159 to a position indicative of full protonation, consistent with the existence of an imidazolium thiolate ion-pair interaction between His-159 and Cys-25 in the active enzyme. PMID- 7470479 TI - Effect of cysteine-25 on the ionization of histidine-159 in papain as determined by proton nuclear magnetic resonance spectroscopy. Evidence for a his-159--Cys-25 ion pair and its possible role in catalysis. AB - Papain was succinylated in order to increase its solubility above pH 8 so that proton NMR spectroscopy could be used to study the ionization of His-159 at the active site of the enzyme. The pH dependence of NMR spectra of catalytically active succinyl-papain and the methylthio derivative of the active-site cysteinyl residue of succinyl-papain (succinyl-papain-S-SCH3) were determined between pH 6 and 10. The pH dependence of the C epsilon 1 H resonance of His-159 in catalytically active succinyl-papain indicates that His-159 has a pK of about 8.6 in the catalytically active form of the enzyme. The position of this resonance in succinyl-papain-S-SCH3 indicates that when the active-site cysteinyl residue is methylthiolated, His-159 is completely deprotonated between pH 6 and 10. This result is taken as evidence for an imidazolium--thiolate ion-pair interaction between His-159 and Cys-25 wherein neutralization of the charge on the thiolate anion by methylthiolation would be expected to cause a marked decrease in the pK of His-159. A possible catalytic role for the ion pair in the acylation step in papain-catalyzed reactions is proposed wherein attack of a substrate by the imidazolium--thiolate ion pair is accompanied by an increase in the acidity of the imidazolium group that facilitates expulsion of the leaving group of the substrate. PMID- 7470480 TI - Perturbations in the free energy and enthalpy of ionization of histidine-159 at the active site of papain as determined by fluorescence spectroscopy. AB - Fluorometric titrations of papain, succinyl-papain, and the corresponding methylthio derivatives of Cys-25 (papain-S-SCH3 and succinyl-papain-S-SCH3) were determined. Removal of the methylthio group from Cys-25 resulted in an increase of approximately 4 pK units in the fluorometrically determined pK value. The correspondence between the ionization behavior as determined by proton NMR and fluorometric titrations indicated that fluorescence titrations reflect the ionization behavior of His-159 in both the active enzyme and the methylthio derivative. The ionic strength dependence of the pK was analyzed in terms of simple electrostatic theory and was shown to be consistent with the charge on the protein. The temperature dependence of the pK values of His-159 indicated an increase in the heat of ionization from about 0 to 8 kcal/mol upon removal of the methylthio blocking group from Cys-25. Measurements of the effect of solvent on the pK's and heats of ionization of simple model compounds indicated that the observed shift in enthalpy of ionization of His-159 upon removal of th methylthio group from Cys-25 is not unreasonable in light of the accompanying perturbation is more than 4 pK units in the pK of His-159. The perturbations in enthalpies and free energies are attributed to formation of an ion pair. The ionization behavior of His-159 in thiol-blocked derivatives of papain is consistent with the involvement of His-159 in the deacylation step in papain catalysis. PMID- 7470482 TI - Chemical modification of the alpha-amino group in snake venom phospholipases A2. A comparison of the interaction of pancreatic and venom phospholipases with lipid -water interfaces. PMID- 7470481 TI - Correlations between deoxyribonucleic acid structural parameters and calculated circular dichroism spectra. AB - Circular dichroism spectra are calculated for 68 different conformations of the bases in the double-stranded sequence of T-G-G-C-G-T-A-T-T-C-T and its complementary strand. All conformations are right-handed helices and include both published models and energy-minimized forms. Examination of the dependence of the circular dichroism band at 274 nm shows that the magnitude of the band is a linear function of both the helix winding angle and the base pair twist. Correlations of the magnitude of the circular dichroism with the distance between base pairs, the distance of the base pairs from the helix axis, or the tilt of the base pair relative to the helix axis are poorer. PMID- 7470483 TI - Hydrogen-1 and phosphorus-31 nuclear magnetic resonance study of the solution structure of Bacillus licheniformis 5S ribonucleic acid. AB - The conformation of Bacillus licheniformis 5S RNA in solution has been studied by using 360-MHz 1H NMR and 40.5-MHz 31P NMR spectroscopy. The 1H NMR spectra, which are well resolved, have been compared with theoretical spectra derived by ring current shift calculations for various models proposed in the literature for the secondary structure of 5S RNA. The total amount of base pairs is estimated to be around 36. NMR melting experiments indicate that both the molecular stalk and the prokaryotic loop [Fox, G. E., & Woese, C. R. (1975) Nature (London) 256, 505] are present in the solution structure. On this basis, some models proposed for the secondary structure of 5S RNA not containing these structural features can be rejected. Several resonances are observed around 10.7 ppm that can be ascribed to protons involved in non-Watson-Crick base pairing most likely present in tertiary interactions in the 5S RNA molecule or to ring N protons of nonpaired bases which as a result of the molecular folding are shielded from the solvent. Under our solution conditions, these structural features disappear at physiological temperature, the process being uncoupled from the collapse of the secondary structure. Using 31P NMR, we demonstrate that the number of phosphate conformations in the sugar phosphate backbone of 5S RNA, deviating from the g-,g- conformation normally found in double helices, is far les than in tRNA. PMID- 7470484 TI - On the purification and mechanism of action of 5-aminoimidazole-4-carboxamide ribonucleotide transformylase from chicken liver. AB - The transformylase from chicken liver catalyzing the formylation of 5 aminoimidazole-4-carboxamide ribonucleotide through the agency of 19 formyltetrahydrofolate has been purified to apparent homogeneity. Inosinicase activity copurifies. This transformylase is not further activated kinetically by the presence of the trifunctional protein in contrast to the glycinamide ribonucleotide transformylase. The enzyme exhibits a greater than 1000-fold preference for the naturally occurring 10-formyltetrahydrofolate cofactor and a sequential reaction pattern. A reinvestigation of the chemical structure of the formylated ribotide product employing 13C and 1H NMR indicated that the imidazole ring remained intact upon formylation, consistent with the originally proposed structure. PMID- 7470485 TI - Metal ion interactions with bovine prothrombin and prothrombin fragment 1. Stoichiometry of binding, protein self-association, and conformational change induced by a variety of metal ions. PMID- 7470486 TI - Analysis of the iron-sulfur cluster of aconitase by natural and magnetic circular dichroism. AB - We have examined the iron-sulfur cluster of aconitase, a high-potential iron sulfur protein, by absorption, circular dichroism (CD), and magnetic circular dichroism (MCD) spectroscopy. The MCD spectrum of unactivated aconitase, which is presumably oxidized, is similar to those of reduced two iron-two sulfide ferredoxins but distinct from the MCD of known four iron-four sulfide proteins. The magnitude of the natural CD of unactivated aconitase also suggests the absence of four iron-four sulfur clusters. Reduction of the enzyme with dithionite and activation with the cysteine-ascorbate-ferrous ion activation mixture generate spectra which are significantly different from those of any iron sulfur protein seen to date. We interpret these results as indicating that aconitase does not contain a four iron-four sulfur cluster generally thought to be characteristic of high-potential iron-sulfur proteins. It could contain a two iron-two sulfur center or some other center such as a cyclic three iron-three sulfur center. PMID- 7470487 TI - Lipids of synaptic vesicles: relevance to the mechanism of membrane fusion. AB - Synaptic vesicles from the electric organ of the marine ray Narcine brasiliensis, purified to at least 90% homogeneity, were analyzed for the lipid and fatty acid content of their membranes. The major lipids (mol %) were phosphatidylcholine (32.3%), phosphatidylethanolamine (20.5%), phosphatidylserine (6.1%), sphingomyelin (3.0%), and cholesterol (33.3%), a composition which did not differ greatly from that of the parent electric organ. While the number of double bonds per fatty acid molecule was similar for both synaptic vesicle and whole electric organ phospholipids, the vesicles were highly enriched in docosahexenoic acid (22:6). Reaction with the amine labeling reagents isethionylacetimidate and trinitrobenzenesulfonic acid indicated that 40% of the phosphatidylserine and 60% of the phosphatidylethanolamine are present on the external (cytoplasmic) surface of the synaptic vesicle. These data on a natural fusing membrane have relevance to models of membrane fusion, which have been based largely on studies of in vitro fusion using synthetic membranes. PMID- 7470488 TI - Arachidonic acid releasing activity in platelet membranes: effects of sulfhydryl modifying reagents. PMID- 7470489 TI - Structural dimorphism of bile salt/lecithin mixed micelles. A possible regulatory mechanism for cholesterol solubility in bile? X-ray structure analysis. AB - The three-dimensional structure of bile salt/lecithin mixed micelles in 0.15 M saline was derived from X-ray small-angle scattering measurements under various conditions. Two essentially different types of micelles were detected. At bile salt:lecithin molar ratios lower than approximately 2:1, lamellar particles similar to a lecithin bilayer arrangement were found. The thickness of the bilayer is 5.1 nm for mixed micelles having a molar ratio of 1:1. The lateral dimensions of the micelle were found strongly dependent upon molar ratio, increasing as lecithin content increases. In addition, it appeared that under certain incompletely defined conditions vesicular particles having diameters in the region of more than 100 nm occur. A molecular model of this type of micelle has been derived by means of a thorough interpretation of the electron density distribution across the plane of the bilayer. This model is generally consistent with the "mixed-disk" model of Mazer et al. [Mazer, N. A., Kwasnick, R. F., Carey, M. C., & Benedek, G. B. (1977) Micellization, Solubilization, Microemulsions, Proc. Int. Symp. 1, 383-402] and thus differs from that proposed by Small [Small, D. M. (1967) Gastroenterology 52,607-610] and Dervichian [Dervichian, D. G. (1968) Adv. Chem. Ser. No. 84, 78-87]. At molar ratios exceeding 2:1, a different type of micelle structure was found. This is a highly isometrical particle of globular shape, probably having a centrosymmetric arrangement of the molecular constituents. At a 3:1 bile salt:lecithin molar ratio, the mean physiological ratio for human gallbladder bile, the diameter of this micelle is 6.2 nm. Provided that other bile constituents have no significant influence on the structure, it may reasonably be expected that native gallbladder bile contains micelles of this latter type. The prevalent balance between the two micellar forms may thus regulate the capacity of bile to transport cholesterol. PMID- 7470490 TI - Dimorphism in bile salt/lecithin mixed micelles. AB - We investigated the dependence of certain properties of bile salt/lecithin mixed micelles on the molar ratio of the two components. UV absorption suggested that the micellar system is fundamentally altered on passing through a bile salt:lecithin molar ratio of 1.8:1. Differential scanning calorimetry confirmed this transition ratio; in addition, it indicated a bilayer arrangement of lecithin, in support of the mixed-disk model of micellar structure, but only for micelles having a molar ratio less than 1.8:1. For micelles having larger ratios, which are those of physiological interest, calorimetry was inconsistent with the mixed-disk model. These observations support the X-ray structure analysis of Muller. PMID- 7470491 TI - Carbon-13 and phosphorus-31 nuclear magnetic resonance studies on interaction of calcium with phosphatidylserine. AB - The interaction between Ca2+ and phosphatidylserine was studied by 13C and 31P NMR spectroscopy, by IR analysis, by binding constant measurements, and through use of space-filling molecular models. NMR measurements of various salt forms of the lipid were made in two types of organic solvents that allowed sufficient averaging of chemical shift anisotropy and dipolar couplings to yield high resolution spectra. 13C resonances of the polar head-group carbons were broadened relative to those of the acyl chains. This was especially true in samples prepared at neutral pH where ionic interactions appeared to restrict molecular motion. In CDCl3 the marked line broadening of the resonances of the polar head group atoms in the Ca2+ form indicated the formation of large, slow tumbling micelles. In the amphipathic solvent the large reduction in line broadening indicated the presence of freely tumbling Ca-(phosphatidylserine)2 dimeric complexes. The 2:1 binding stoichiometry and the low chemical activity of the Ca phosphatidylserine complex support this view. Analysis of the chemical shifts of the various lipid atoms under the differing ionic environments indicates that Ca2+ enhanced the deprotonation of both the carboxyl and amino groups and stabilized the entire polar head group against the effects of changing pH. The marked upfield shift of the 31P phosphate resonance in the Ca2+ form and its insensitivity to changing pH indicate strong coordination binding. IR data indicate direct involvement of the carboxyl group in Ca2+ binding, as evidenced by the appearance of a C=O stretching mode. Binding studies indicated that the phosphate group was the primary binding force but that the carboxyl group also contributes positively. The amino group appears to exert a repulsive effect, which is supported by the chemical shift data which indicate that Ca2+ enhances the deprotonation of the amino group. Molecular models indicate direct involvement of the carboxyl and phosphate oxygens and that the amino group must be deprotonated to participate. PMID- 7470492 TI - Photochemical and functional properties of bacteriorhodopsins formed from 5,6 dihydro- and 5,6-dihydrodesmethylretinals. AB - 5,6-Dihydroretinal and 5,6-dihydro-1,1,5,9,13-desmethylretinal are synthesized, and their all-trans isomers are shown to form pigment analogues (lambda max at 475 and 460 nm, respectively) of bacteriorhodopsin (purple membrane protein). The shift of the absorption maximum od the pigment from that of the protonated Schiff base of the chromophore for 5,6-dihydrobacteriorhodopsin is small compared to that of the native pigment, suggesting that negative charges similar to those controlling the lambda max of visual pigment rhodopsin exist near the cyclohexyl ring. Both pigment analogues undergo reversible light-induced spectral shifts reflecting cyclic photoreactions of the pigments. These results indicate that the absence of the C-5--C-6 double bond and of the five methyl groups of retinal does not abolish the photochemistry of these pigment analogues and strongly suggest that these structural features are not directly required for the photoreactions of native bacteriorhodopsin. The apparent rates of the photochemical transformations of these artificial pigments are quite different from those of bacteriorhodopsin. A working hypothesis is proposed for the photocycle of the pigment analogues, which includes a slower light-induced cycling rate (for the light-adapted pigments) than that of native bacteriorhodopsin and an increased rate of dark adaptation. When incorporated into egg lecithin vesicles both pigment analogues show proton pumping ability, again indicating that the missing double bond and the methyl groups are not structurally required for the function of the pigments. PMID- 7470493 TI - Fragmentation of proteins with o-iodosobenzoic acid: chemical mechanism and identification of o-iodoxybenzoic acid as a reactive contaminant that modifies tyrosyl residues. AB - o-Iodoxybenzoic acid, a disproportionation product of o-iodosobenzoic acid, has been identified as a contaminant in most preparations of o-iodosobenzoic acid capable of both modifying and cleaving certain tyrosyl residues. A new synthetic approach for the production of o-iodosobenzoic acid containing low amounts of o iodoxybenzoic acid combined with preincubation of the reagent with p-cresol to destroy the remaining o-iodoxybenzoic acid prior to the reaction with a polypeptide allows preparation of reagent solutions in which tyrosyl residues remain intact during tryptophanyl bond cleavage. In addition, the product produced by the action of o-iodosobenzoic acid upon tryptophanyl bonds has been identified as N-acyldioxindolylalanine. It is inferred from that structure that the chemical reaction proceeds via a two-step oxidation of the tryptophanyl residue followed by formation of an iminospirolactone which hydrolyzes, cleaving the peptide chain. Small peptides ending with dioxindolylalanine can be coupled to aminopropyl glass in high yield and are suitable for solid-phase Edman degradation. PMID- 7470494 TI - Evidence for a plasma membrane redox system on intact ascites tumor cells with different metastatic capacity. AB - A NADH-ferricyanide reductase of the external surface of intact mouse ascites tumor cells grown in culture was shown. The oxidation/reduction reaction was due to enzymatic rather than inorganic iron catalysis as demonstrated by the kinetics and specificity of the reaction. Activities of three markers for cytoplasmic contents were lacking with the intact tumor cells. The dehydrogenase activity was inhibited by p-chloromercuribenzoate, bathophenanthroline sulfonate, and the anticancer drug adriamycin. Sodium azide and potassium cyanide inhibited partially. The response to inhibitors resembled that of isolated plasma membranes rather than that of mitochondria. Concurrent with these findings, neither superoxide dismutase nor rotenone affected the redox activity. The findings provide evidence for the operation of a plasma membrane redox system at the surface of intact, living cells. PMID- 7470495 TI - A comparison of the phosphorylation potential and electrochemical proton gradient in mung bean mitochondria and phosphorylating sub-mitochondrial particles. AB - The phosphorylation potential (delta Gp) and the electrochemical proton gradient (delta muH+) normally maintained during respiration or ATP hydrolysis by mung bean hypocotyl mitochondria and phosphorylating sub-mitochondrial particles have been investigated. Phosphorylation potential experiments using safranine and oxonol-VI, as membrane potential markers for mitochondria and sub-mitochondrial particles, respectively, suggest that the 'null point' delta Gp (i.e., the phosphorylation potential at which no change in optical signal occurred) corresponds to a value of 15.2 +/- 0.7 kcal/mol in mitochondria and 11.2 +/- 0.3 kcal/mol in sub-mitochondrial particles. The value of delta muH+ generated by the hydrolysis of ATP was estimated using ion distribution techniques. In each case a rapid centrifugation technique was used to separate the organelle from the suspending medium. The total delta muH+ generated in each case was approx. 200 mV being composed of both membrane potential and pH components. A comparison of delta muH+ with delta Gp indicates that the apparent H+/ATP ratio in mung bean mitochondria is 3.4 +/- 0.2 while in phosphorylating sub-mitochondrial particles it is 2.2 +/- 0.1. PMID- 7470496 TI - Photosynthetic energy conservation investigated by thermoluminescence. Activation energies and half-lives of thermoluminescence bands of chloroplasts determined by mathematical resolution of glow curves. AB - Thermoluminescence of isolated chloroplasts was analysed by a computer-assisted multicomponent curve fitting procedure to determine the activation energies, the free energies of activation, frequency factors and half-lives of the component bands of the glow curve. Optimal fit was obtained in the temperature region from 80 degrees C to +80 degrees C by the resolution of the glow curve into seven bands with peak positions at -24, -12, +12, +17, +28, +44, and +69 degrees C. All of the activation free energies of the thermoluminescence bands were much higher than 0.59 eV, the minimum free energy of activation required for the back reaction of the primary charge separation as calculated on the basis of the theory of Ross and Calvin (Ross, R.T. and Calvin, M. (1967) Biophys. J. 7, 595 614). The high free energies of activation and long half-lives (longer than 50 ms) of the thermoluminescence bands suggest that thermoluminescence in the temperature region from -80 degrees C to 80+ C does not reflect the change recombination of primary products but represent the reversal of subsequent stabilization steps of the charge separation process which proceed along the acceptor and donor sides of Photosystem II. PMID- 7470497 TI - Experimental and theoretical considerations of mechanisms controlling cation effects on thylakoid membrane stacking and chlorophyll fluorescence. AB - The roles of specific cation binding, charge neutralization and electrostatic screening mechanisms in controlling salt-induced stacking and chlorophyll fluorescence changes in thylakoid membranes are examined in the light of new experimental evidence and theoretical calculations of the forces between membrane surfaces. A comparison of the biphasic stacking and fluorescence phenomena generated by organic mono- and divalent cations known sterically to inhibit specific binding with the effects generated by inorganic mono- and divalent cations suggests that the observed salt-induced changes at pH greater than or equal to 7.5 are predominantly governed by the electrostatic screening mechanism in agreement with previous work (e.g. Barber, J., Mills, J.D. and Love, A. (1977) FEBS Lett. 74, 174-181). Detailed calculations of the coulombic double layer repulsive force between negatively charged membrane surfaces immersed in a mixed electrolyte of valence type Z1+/Z1-,Z2+/Z1- were performed both under the constraints of fixed surface charged density and fixed surface potential. From a close comparison of the theoretical results with new experimental data on salt induced stacking and fluorescence changes and a consideration of the contributions of the 'hydration' repulsive force and the van der Waals attractive force, it is argued that a reduction in surface charge density alone by lateral diffusion is probably insufficient to realize membrane stacking and that an increase in the van der Waals attractive force is necessary to account for the experimental observations perhaps through the formation of protein rich domains. In view of the complexity of the thylakoid membranes, the conclusions are to be considered qualitative. Nevertheless, these calculations give support to a model in which the cation induced chlorophyll fluorescence and stacking changes can be explained by lateral diffusion of two types of pigment protein complexes in the lipid matrix of the membrane. Such diffusion gives rise to changes in energy transfer between Photosystem II and Photosystem I and also to the creation of domains having low and high electrical surface charge density. PMID- 7470498 TI - Uniparental inheritance of a chloroplast photosystem II polypeptide controlling herbicide binding. AB - The ability of atrazine to inhibit Photosystem II electron transport and the rate of electron transfer from the primary to the secondary quinone electron acceptors in the photosystem II complex were examined in triazine-resistant and susceptible parental biotypes of Brassica campestris L. and their F1 progeny derived from reciprocal crosses. The lack of herbicide inhibitory activity and the presence of functional properties which decreased the Q- to B electron transport rate constant were inherited in parallel through the maternal parent. We conclude that the herbicide receptor protein is uniparentally inherited through the female parent. These data are discussed in relation to other studies which indicate that the binding site is a 32 000-dalton polypeptide which determines the functional properties of B (the secondary Photosystem II electron acceptor). PMID- 7470499 TI - Characterization of a new type of ferredoxin from Desulfovibrio africanus. AB - A new ferredoxin designated ferredoxin III has been isolated from Desulfovibrio africanus grown on media high in iron. Native ferredoxin III is a dimer constituted by two identical subunits of approx. 7500. It is distinguished from the two other ferredoxins (I and II) isolated from this microorganism by its amino acids composition, N-terminal sequence, spectral properties and iron-sulfur content. The amino acid composition of D. africanus ferredoxin III is typical of ferredoxins with an excess of acidic over basic residues and the absence of histidine and arginine residues. The absorption spectrum of ferredoxin III exhibits two maxima, at 408 nm (epsilon = 58.5 . 10(3) M-1 . cm-1) and 285 nm (epsilon = 82 . 10(3) M-1 . cm-1), with a shoulder at 305 nm (epsilon = 75 . 10(3) M-1 . cm-1). Its A408/A285 absorbance ratio is 0.78. Ferredoxin III contains approx. 12--13 atoms each of iron and labile sulfur. This is in agreement with the high value of the extinction coefficient at 408 nm, which is slightly higher than 3-fold that of one [4Fe-4S] cluster. However, the number of cysteine residues of the protein (six residues), which is about the half that of iron atoms, is indicative of the presence of a new type of iron-sulfur cluster in ferredoxin III. The protein is unstable in a low ionic strength environment; the addition of neutral salts stabilizes the protein conformation. The data on the biological activity of ferredoxin III as compared to the two other ferredoxins from D. africanus show that the three iron-sulfur proteins function with equal effectiveness as electron carrier in the phosphoroclastic reaction and the H2 sulfite reductase system. PMID- 7470500 TI - Mitochondrial calcium transport and calcium-activated phospholipase in porcine malignant hyperthermia. AB - The interaction of Ca2+ with mitochondria isolated from longissismus dorsi, a predominantly white skeletal muscle, of normal and malignant hyperthermia pigs was investigated using tightly-coupled preparations. Arrhenius plots of mitochondrial Ca2+ -stimulated respiration for succinate oxidation of malignant hyperthermia pigs showed a transition temperature (Tt) of 26.31 +/- 0.80 degrees C (n = 5), which was decreased by spermine to 15.41 +/- 0.69 degrees C (n = 3), a value very similar to that for normal pigs. No difference in either the Tt or in the activation energy (Ea) was observed between the two types of pigs when ADP was used instead of Ca2+. Mitochondria of malignant hyperthermia pigs were uncoupled at 40 degrees C by exogenous Ca2+ at 1221 +/- 301 (n = 9) nmol Ca2+ per mg proteinn during succinate oxidation and the uncoupled mitochondria showed large amplitude swelling. Both the Ca2+ -induced uncoupling and swelling were prevented by bovine serum albumin and by the phospholipase inhibitors, spermine and tetracaine. In contrast, mitochondria of normal pigs were still tightly coupled even after a total addition of 2313 +/- 287 (n = 5) nmol Ca2+ per mg protein and retained the original condensed configuration in the presence or absence of spermine and tetracaine. Mitochondria of malignant hyperthermia pigs contained significantly (P less than 0.001) higher quantities of endogenous Ca2+ and showed a significantly (P less than 0.001) faster FCCP-induced endogenous Ca2+ efflux rate than normal when monitored spectroscopically with murexide. No significant difference was observed in either the rate of exogenous Ca2+ uptake or in the extent of Ca2+ accumulated in the aerobic steady state during succinate oxidation between the two types of pigs. The rate of mitochondrial Ca2+ efflux of malignant hyperthermia pigs during anaerobiosis was about twice that of normal. Experimental evidence suggests that mitochondria from musculi longissimus dorsi of malignant hyperthermia pigs contained a Ca2+ -stimulated phospholipase A2 (EC 3.1.1.4, phosphatide 2-acylhydrolase), and that this enzyme if present in mitochondria of normal pigs is either latent or in very low concentration. The significance of the Ca2+ -stimulated phospholipase A2 and its association with the enhanced rate of glycolysis in porcine malignant hyperthermia syndrome and in the post-mortem formation of the pale, soft and exudative conditions observed in white skeletal muscles of malignant hyperthermia pigs is discussed. PMID- 7470501 TI - Midpoint potentials of cytochromes in vesicles of anaerobically-grown Paracoccus denitrificans determined by the indirect coulometric titration method. AB - 1. Multiplicity of redox components with spectral properties similar to b-type cytochromes was established in vesicles derived fro anaerobically-grown Paracoccus denitrificans. 2. Multiplicity of c-type cytochromes was not apparent either from low temperature spectroscopy or potentiometric titrations. 3. Cytochromes a + a3 and a component, only observable at liquid nitrogen temperature, with a spectral maximum at 582.5 nm were detected. 4. Redox cycling of electron transport components using the indirect coulometric titration method was a convenient means of pairing redox potentials and was reproducible in total absorbance changes, midpoint potentials and spectral maxima. PMID- 7470502 TI - Charging of the A band of thermoluminescence, dependent on the S3 state in isolated chloroplasts. AB - The intensity of the A band of thermoluminescence from isolated spinach chloroplasts were greatly dependent on the excitation temperature between -40 and -10 degrees C. This dependency was analyzed in relation to the S states in the oxygen-evolving system, and the following results were obtained. 1. The A band was charged only slightly by excitation at low temperature -65 degrees C) with continuous light, but when two flashes had been given before cooling the A band was charged efficiently by the low temperature excitation. 2. The efficiency of the double flash pre-illumination for enhancing the A band showed a steep temperature dependency which agreed with the temperature dependency of the photoconversion from the S2 to S3 state. 3. The A-band-enhancing effect was dependent on the number of flashes, and underwent a quadruple oscillation. 4. The enhancing effect decayed in darkness and the decay course agreed with that of the B1 band but not with that of the B2 band. From these results, the origin of positive charges for the A band was discussed. PMID- 7470504 TI - Salt-induced microscopic changes in chlorophyll fluorescence distribution in the thylakoid membrane. AB - Addition of 3 mM MgCl2 to isolated pea thylakoids suspended in a medium of low osmotic strength at room temperature induces an increase in chlorophyll fluorescence similar to that observed with unswollen thylakoids. Fluorescence microscopy indicates that the MgCl2 induced increase in the emission intensity involves the formation of highly fluorescent patches on the swollen vesicles. The data seems to give additional support ton the concept that salt induced chlorophyll fluorescence changes involves the lateral movement of pigment proteins within the thylakoid membrane in such a way as to form discrete domains. PMID- 7470503 TI - On the active species in bicarbonate stimulation of Hill reaction in thylakoid membranes. AB - At 5 degree C addition of CO2 or HCO3- to CO (HCO3-)-depleted thylakoids (containing 100 mM formate) initiates, within 10 s, the activation of the Hill reaction in light. In contrast to HCO3- addition, where there is a lag of 6-8 s, the activation by CO2 addition is almost instantaneous. With CO2, prior addition of carbonic anhydrase produces a lag of about 6 s that approaches the lag observed by the HCO3- addition. These data suggest that CO2 is an active species involved in stimulating the Hill reaction. Binding of CO2 (HCO3-) to a component on the external side of the thylakoid membranes is supported by the present study. PMID- 7470505 TI - Characterization of a proteoglycan of high electrophoretic mobility. AB - Proteoglycans from articular cartilage of young baboons (Papio papio) were fractionated on an associative density gradient. Material from the top of the gradient was shown to contain a proteoglycan of high electrophoretic mobility on large porosity gels. Associated and/or contaminating proteins were removed by ion exchange chromatography on DEAE-cellulose and subsequent gel filtration on Sepharose 4B in the presence of 0.1% SDS. The electrophoretically homogeneous proteoglycan (Kd 0.43 on Sepharose 4B SDS) contained 39.7% protein, was rich in aspartate, glutamate, leucine and glycine and had a GalN : GluN molar ratio of 3.87. PMID- 7470506 TI - Role of guanine nucleotides in the stimulation of thyroid adenylate cyclase by prostaglandin E1 and cholera toxin. AB - Cholera toxin in the presence of GTP increased adenylate cyclase activity in a purified bovine thyroid plasma membrane preparation, whereas, in the presence of guanosine 5'-(beta, gamma-imido)-triphosphate (Gpp(NH)P), cholera toxin had no stimulatory effect. Similarly, prostaglandin E1 enhanced the adenylate cyclase activity induced by GTP but not by Gpp(NH)p. Gpp(NH)p-stimulated adenylate cyclase activity, assayed with hydrolysis-resistant adenosine 5'-(beta, gamma imido)-[32P]triphosphate as substrate and no ATP-regenerating system was inhibited by GDP in a competitive fashion. Furthermore, prostaglandin E1, but not cholera toxin, influenced the GDP inhibition of Gpp(NH)p-stimulated activity by increasing the concentration of GDP resulting in 50% inhibition approx. 2-fold. Inosyl nucleotides mimicked the effects of guanyl nucleotides on thyroid adenylate cyclase in that ITP could substitute for GTP in enhancing cholera toxin and prostaglandin #1-induced activities and that inosine 5'(beta, gamma-imido) triphosphate [Ipp(NH)p] was also a potent stimulator per se. Conclusions. (1) Cholera Toxin and prostaglandin E1 enhance thyroid adenylate cyclase activation by GTP (or ITP), but have no stimulatory effect on the Gpp(NH)p (or Ipp(NH)p) response; (2) the stimulatory effect of prostaglandin E1 on adenylate cyclase may result from decreased affinity for GDP at the guanine nucleotide regulatory site; (3) the date regarding cholera toxin stimulation of thyroid adenylate cyclase are consistent with the hypothesis that cholera toxin exerts its effect by inhibiting an endogenous GTPase. PMID- 7470507 TI - Comparison of mitostatic effect, cell uptake and tubulin-binding activity on colchicine and colcemid. AB - Mitostatic action, cellular uptake and the binding of colchicine and colcemid to tubulin were compared. It was shown that mitostatic action of low doses of colchicine developed only after 24 h incubation of the drug with mouse L fibroblasts, while the colcemid-induced block of mitosis was evident after 2 h incubation. The initial rate of uptake was about 10 times greater for colcemid than for colchicine. Cellular uptake of the drugs reached an equilibrium after 2 and 15-18 h incubation for colcemid and colchicine, respectively, and the plateau values were identical. The kinetics of colchicine and colcemid binding to bovine brain tubulin was studied by the DEAE-filter binding assay. Colcemid binds to tubulin much faster than does colchicine. The rate of colcemid efflux from L cells is much higher than that of colchicine. According to the efflux data, colcemid dissociates readily from a complex with tubulin (t1/2 = 10 min), while the colchicine-tubulin complex is stable for at least 1 h. These results are consistent with previously published data (Frankel, F.R. (1976) Proc. Natl. Acad. Sci. U.S.A. 72, 2798-2802), which showed that colcemid action on cells is more reversible than that of colchicine. We suggest that differences between colchicine and colcemid in the rate of mitostatic action and its reversibility are determined by the differences in parameters of tubulin binding. PMID- 7470508 TI - Studies on the relative rates of intracellular transport of egg white proteins. AB - In order to determine if the rates of intracellular transport of oviduct secretory proteins are coordinate, oviduct minces were first incubated with either [3H]methionine or [3H]leucine. 2 h later, the minces were exposed to either [35S]methionine or [14C]leucine in addition to the 3H-labeled amino acid. After incubating for an additional 90-120 min, aliquots of the media were subjected to immunoprecipitation to isolate ovalbumin, conalbumin and lysozyme. From the 3H/35S or 3H/14C ratios in the isolated proteins, relative rates of intracellular transport were determined. The data obtained indicate that conalbumin and lysozyme exhibit similar rates of intracellular transport, whereas ovalbumin lags behind these proteins considerably. The basis for this effect is discussed in light of current knowledge concerning the synthesis and secretion of ovalbumin. PMID- 7470509 TI - Purification and characterization of Treponema hyodysenteriae hemolysin. AB - A hemolysin produced by Treponema hyodysenteriae ATCC27164 was purified from broth filtrates by acetic and (NH4)2SO4 precipitations followed by ion exchange chromatography on diethylaminoethyl-Sephacel and gel filtration using Ultrogel AcA44. The purified hemolysin displayed only one band on polyacrylamide gel electrophoresis. By gel filtration the molecular weight was estimated as 74,000 daltons. The isolated hemolysin was oxygen resistant, heat labile and was not inactivated over a wide range of pH values. Further analysis indicated that this hemolysin was probably a polypeptide or a protein associated with lipids and nucleotides. Its action on rabbit erythrocytes which did not require any divalent cations could not be related to a lipolytic or proteolytic activity. PMID- 7470510 TI - Characteristics of the interaction of a treponemal hemolysin with rabbit erythrocytes. AB - The mechanism of action on rabbit red cells of Treponema hyodysenteriae hemolysin was studied using volume analysis and release of hemoglobin. While fixation of the hemolysin on the erythrocytes is temperature independent, it appears that hemolysis is temperature dependent. The kinetics of hemolysis proceed according to a sigmoid curve characterized by a prelytic lag. The duration of the prelytic lag varies inversely with the quantity of hemolysin but the rate and the maximum value of hemolysis are directly proportional to the quantity of hemolysin. The effect of sucrose and trypan blue on the hemolysin and the the red cells suggest that erythrocyte lysis is likely to be induced by the hemolysin in a way different from that known for other hemolytic agents. PMID- 7470511 TI - Absence of phosphates from the myosin heavy chains of striated muscles. AB - This work aimed to determine whether the heavy chains of myosin from different striated muscle were phosphorylated. Myosin and its heavy chains were prepared from cardiac and skeletal muscles of rats injected in vivo with radioactive phosphates. The results for radioactive phosphate localization indicate the absence of phosphate from pure heavy chains and from any of their purified fragments, whatever the striated muscle used. In addition, phosphates are present in the myosin phosphorylated light chain and in a contaminating protein closely associated to the myosin heavy chain. PMID- 7470512 TI - Metabolism of glycosaminoglycans in cultured smooth muscle cells from pig aorta. PMID- 7470514 TI - One-step preparation of purified plasma membranes from pigeon erythrocytes. PMID- 7470513 TI - [Inhibition by the high density lipoprotein HDL2 and HDL3 of DNA and sterol biosynthesis in human lymphocytes stimulated with concanavalin A]. AB - Lipoproteins HDL2 and HDL3 inhibit DNA synthesis and sterol synthesis in human Con A-stimulated lymphocytes cultured in a medium supplemented with 20 per cent lipoprotein deficient serum. On the basis of the amount of proteins added, HDL2 is more efficient on DNA and sterol synthesis than HDL3 and less efficient than LDL. However, on the basis of the amount of cholesterol added, the inhibition of sterol synthesis induced by these three lipoproteins is not significantly different. At all concentrations of these three lipoproteins, the inhibition of sterol synthesis is higher than the inhibition of DNA synthesis. PMID- 7470515 TI - Individual differences in alpha rhythm responsivity: inter-task consistency and relationships to cardiovascular and dispositional variables. AB - Individual differences in EEG alpha rhythm responsivity were examined with respect to: (1) consistency of response across cognitive tasks; (2) relationships to cardiovascular responsivity; and (3) relationships to dispositional variables. Inter-task correlations revealed a significant degree of stability in subjects' alpha responses to four "sensory rejection" tasks: mental arithmetic, time estimation and imagining oneself as cold or warm. Individuals evidencing greater suppression of alpha activity during mental arithmetic also displayed greater heart rate increase and greater digital vasoconstriction, as well as obtaining higher scores on the Activity subscale of the EASI-III Temperament Survey and on the "Hard Striving" subscale of the Jenkins Activity Survey for Health Prediction, a measure of the Coronary-Prone Behavior Pattern. Alpha responsivity in the time estimation and warm imagery tasks was also associated with greater heart rate increase. The results suggest that alpha responsivity is related to individual differences affecting one's attentiveness or involvement in experimental tasks. PMID- 7470516 TI - The effect of repetition of infrequent familiar and unfamiliar visual patterns on components of the event-related brain potential. AB - This experiment examined changes in the waveforms of the event-related brain potential (ERP) during repeated presentations of infrequent-familiar and infrequent-unfamiliar visual patterns. The EEG waveforms were averaged separately for each presentation of the two types of stimuli across different stimulus blocks. Principal components analysis and baseline-to-peak measurement identified three ERP components, a P3, P4 and a slow wave of diminishing positivity at anterior, central and posterior scalp sites. Unfamiliar stimuli evoked components of greater positivity and P3 waves of longer latencies than familiar stimuli. Repetition of these stimuli was associated with a progressive diminution of the amplitude of the three components and a shortening of P3 latency. For the slow wave the decline in amplitude was more pronounced during repetition of unfamiliar than familiar stimuli. These effects are interpreted as signs of alterations in cognitive processes that play a role during orienting and perceptual learning. PMID- 7470517 TI - Physiologic correlates of direct and averted gaze. AB - The present study was designed to explore the effects of reciprocated and unreciprocated gaze upon skin conductance and heart rate response. A paradigm used previously to investigate maternal response to infant gaze was selected which allowed us to consider possible order effects, sex differences and differential rates of habituation in response to direct and averted gaze. Twenty four males and 24 females were shown 10 sec silent images of an adult face on a videotape monitor of an adult face. In one condition, the en face position offered eye contact; in the other condition, eye contact was not possible. Each subject viewed a sequence of six identical episodes of one condition followed by six episodes of the other condition. Analysis of skin conductance response revealed a significant effect of stimulus sex (p less than 0.05) with the male stimulus figures eliciting the greater skin conductance response. HR responses to the two conditions varied not only as a function of stimulus sex but also as a function of the viewer's sex. The most distinctive data came from a male viewing another male. For this group alone, responding was consistent to unreciprocated gaze viewed first (p less than 0.01) and viewed second (p less than 0.05); and for this group alone responding to reciprocated gaze viewed first habituated (p less than 0.05). PMID- 7470518 TI - Voluntary heart rate lowering following a cardiovascular arousing task. AB - The purpose of the research was to evaluate the effect of feedback-assisted voluntary control to lower heart rate (HR), following a non-somatic mediated task designed to induce cardiovascular activation. Twenty-seven male subjects participated in this study and were randomly divided into three groups. All subjects were submitted to an arousing mental arithmetic task for a 1 min period, followed either by a feedback-assisted HR lowering task (FG), by an unassisted HR lowering task (ICG), or by a relaxation period (RCG). During this period (2 min duration), ICG and RCG subjects were required, as a help for HR deceleration or relaxation, to track visually a sweeping line similar in form to the continuous analogue feedback of the FG subjects. This sequence, interspaced by a 1 min rest period, was repeated 10 times. Results showed that the mental arithmetic task was effective in producing consistent and repetitive HR acceleration. All three groups showed significative HR lowering during the following voluntary HR control or relaxation period. However, there was no difference in the speed or depth of HR lowering. It appeared that continuous-analogue feedback was not more effective than instructions to relax to assist HR return to lower levels. PMID- 7470519 TI - Electrodermal reactions to opposite types of autogenic training imagery. AB - An experiment was conducted to determine the effects of autogenic and "opposite autogenic" verbal formulae upon electrodermal activity. In a between-subjects design, 15 unpracticed volunteers listened to and followed tape recorded imagery suggestions for relaxation and quietude, heaviness and warmth in the extremities, warmth in the central area of the body, coolness of the forehead, and calmness and regularity of breathing and circulation; 15 additional volunteers were exposed to opposite suggestions. Frequency of spontaneous GSRs, cumulative peak amplitude of spontaneous GSRs, and basal skin resistance levels were measured during 2 min epochs before and after the suggestions. Directionally appropriate changes occurred in all three electrodermal measures; these changes were significant for the two GSR measures and approached significance for the BSR measure. The autogenic suggestions produced greater and more reliable visceral effects than did the "opposite autogenic" suggestions. PMID- 7470520 TI - Reliability of spontaneous electrodermal activity in the cat as a function of waking and sleep stages. AB - This study was designed to examine the reliability of spontaneous electrodermal activity (EDA) as a function of the stages of sleep and waking in freely moving cats. Five adult cats were observed during waking--sleep sessions. The results show that: (a) for all stages, the reliability of EDA is slightly higher for amplitude of SSPRs than for frequency; (b) during drowsiness, a maximum of reliability is observed, as is a slight decrease during slow wave sleep; during paradoxical sleep, reliability decreases greatly to below that of the waking level; (c) the reliability of spontaneous EDA appears to be higher in waking cats than that quoted for human subjects. These results are discussed with reference to individual characteristics and state variables. PMID- 7470521 TI - Early auditory information processing in schizophrenia: a preliminary report. AB - Numerous theorists have postulated that incoming environmental information is altered with schizophrenic illness, due to aberrant sensory processing in subcortical systems. Brainstem evoked potential recordings were obtained during passive auditory information processing in groups of chronic schizophrenic patients, affective disordered patients, and normals. These neuroelectric measures of brainstem and midbrain functioning suggest that the three study groups do not statistically differ in terms of latency characteristics. It is noteworthy, however, that a particular subset of chronic schizophrenic patients yielded latency characteristics falling more than one standard deviation from the total population mean under all experimental conditions. PMID- 7470522 TI - Postpartum estrus in Norway rats. II. Physiology. PMID- 7470523 TI - Influence of injected selenium in dairy bulls on blood and semen selenium, glutathione peroxidase and seminal quality. PMID- 7470524 TI - Comparison of serum progesterone, 20 alpha-dihydroprogesterone, and estradiol-17 beta in pregnant and pseudopregnant rabbits: evidence for postimplantation recognition of pregnancy. PMID- 7470525 TI - Uterine contractions and electrical activity in ewes treated with selenium and vitamin E. PMID- 7470526 TI - Episodic fluctuation of total estrogens in peripheral blood of bulls: effects of estradiol-17 beta on LH and testosterone secretion. PMID- 7470527 TI - Dolichol concentration and biosynthesis in rat testis and epididymis. PMID- 7470528 TI - Relaxin improves the coordination of rat myometrial activity in vivo. PMID- 7470529 TI - Reproduction in domestic and feral swine. II. Interrelationships between fetal size and spacing and litter size. PMID- 7470530 TI - Induced protein and deciduoma formation in rat uterus. PMID- 7470531 TI - Energy requirements of the developing mammalian blastocyst for active ion transport. PMID- 7470532 TI - Estrogen and uterine sensitization for the decidual cell reaction: role of prostaglandins. PMID- 7470533 TI - Sequential inhibitory effect of progesterone on lordotic responsiveness in rats: time course, estrogenic nullification, and actinomycin-D insensitivity. PMID- 7470535 TI - Postpartum estrus in Norway rats. I. Behavior. PMID- 7470534 TI - Concentrations of seven elements in the intraluminal fluids of the rat seminiferous tubules, rate testis, and epididymis. PMID- 7470536 TI - Estradiol, testosterone, and androstenedione in ovine follicular fluid during growth and atresia of ovarian follicles. PMID- 7470537 TI - Release of uterine contraction-inducing factor from preovulatory follicle. PMID- 7470538 TI - Effects of maternal undernutrition on the development of the ovine fetus and the associated changes in growth hormone and prolactin. PMID- 7470539 TI - Daily luteinizing hormone release in ovariectomized hamsters: effect of barbiturate blockade. PMID- 7470540 TI - Pregnancy-associated murine protein 3 (pamp-3): characterization and serum levels. PMID- 7470541 TI - Differential stimulation of uterine cells by nafoxidine and clomiphene: relationship between nuclear estrogen receptors and type II estrogen binding sites and cellular growth. PMID- 7470542 TI - Infertility in mice exposed in utero to benzo(a)pyrene. PMID- 7470543 TI - Chicken and turkey spermatozoal superoxide dismutase: a comparative study. PMID- 7470544 TI - Fate of acrosomal glycoproteins during the acrosomal reaction and fertilization: a light and electron microscope autoradiographic study. PMID- 7470545 TI - Temporal patterns of neural activity and their relation to the secretion of posterior pituitary hormones. PMID- 7470546 TI - Electrochemical characteristics of the S.I.A. dental amalgam cavity liner. AB - Selective interfacial amalgamation (S.I.A.) has been developed as a dental amalgam cavity liner with the intent of increasing the restoration-tooth adhesion and reducing the debilitating effects of corrosion and marginal leakage. This paper examines the S.I.A. liner's effectiveness as a corrosion preventive cavity liner by two experimental techniques: anodic polarization and differential aeration tests. The results from these tests indicate that S.I.A. does enhance the corrosion resistance of a dental amalgam. PMID- 7470547 TI - Controlled release of multi-component cytotoxic agents from radiation polymerized composites. AB - Multi-component cytotoxic (anticancer) agents such as mitomycin C (MMC), adryamycin (ADM) and 1-(2-tetrahydrofuryl-5-fluorouracil) (FT-207), were entrapped in a single common composite by radiation-induced polymerization of glass-forming monomers in the presence of polymers. The release profiles of each cytotoxic agent were controlled by the contents and compositions of three cytotoxic agents in the matrix. The release rates of each cytotoxic agent were retarded by addition of an adsorbent and accelerated by addition of a pore-making agent. The release fom the common matrix of a cytotoxic agent and its promoter which have markedly different molecular weights [e.g. MMC and urokinase (UK)] was also investigated. A double entrapping method was successfully used for controlled release in such cases. In conclusion it was found that the release profiles of multi-components in a single matrix could be controlled by using these techniques. PMID- 7470548 TI - Influence of Collagenfleece on bone regeneration. AB - It can be demonstrated that bone regeneration is stimulated by implantation of Collagenfleece a purified, heterologous collagen preparation. Defects were created in rabbit mandibles which were filled with Collagenfleece on the right side, while the defect on the left side served as a control. The progress in bone regeneration was followed by comparative light and electron microscopic investigations. In contrast to the controls, the defects which were treated with Collagenfleece showed quicker ossification. PMID- 7470549 TI - Properties and applications of Bioplast, an absorbable surgical implant material from fibrin. AB - Bioplast is the only commercially available absorbable implant material. Compression moulded from fibrin, it is a yellowish, translucent and flexible material which is thermally stable below 170 degrees C. Its mechanical strength depends on glycerol content. Crosslinking with formaldehyde reduces swelling and digestibility. Bioplant is non-antigenic and capable of being absorbed by polymorphonuclear digestive action. The non-toxic metabolites are eliminated via the kidneys while the site is invaded by host tissue. Clinical applications are mentioned. PMID- 7470550 TI - Problems of encapsulation of total joint replacements. AB - Particulate wear debris of artificial joints, metallic and plastic, and solutions of metals in body fluids are known to have adverse effects. Encapsulation of the articulating parts would prevent wear debris from entering the surrounding tissue and check tissue ingrowth into the joint. The capsule could contain an artificial lubricant to reduce friction and wear and increase the joint life. For the capsule material, several bioelastomers are being tested in an environment of body fluids and lubricants. Three fatigue rigs have been developed for these tests. Tests have shown that a bovine synovial fluid environment causes a tenfold increase in the fatigue life of silicone rubber. A special capsule shape for minimizing stresses due to creasing is being investigated. Electrophoresis has been explored as a method of filtration of wear debris. It has been shown that Co Cr-Mo alloy wear particles are attracted to polymers having permanent negative surface charge. PMID- 7470551 TI - Carbon fibre reinforced epoxy as a high strength, low modulus material for internal fixation plates. AB - A multi-axial carbon fibre/epoxy composite fracture plate has been developed. This type of plate appears to allow early consolidation of internally fixed fractures whilst avoiding the ensuing disadvantage of 'plate induced osteopoenia' which are observed with metal devices. The mechanical properties of these plates (determined 'in vitro') are reported, along with early results of implantation in sheep. PMID- 7470552 TI - Bioceramics consisting of calcium phosphate salts. AB - After a discussion of the chemical considerations involved in the production of calcium phosphate ceramics, animal and clinical studies are reviewed. Specific conclusions are that although these ceramics have high compressive strengths, they are not suitable for joint replacements. Prevention of alveolar ridge resorption is a promising concept, and space-filling and use of aids for jaw-bone enlargement are proposed for routine clinical application. PMID- 7470553 TI - Fatigue test machine for denture base polymers. AB - A description of a single station fatigue test machine suitable for denture base polymers is given. This machine has been especially designed for dental purposes. A group of representative denture base polymers has been tested, which includes dental poly(methylmethacrylate) of the standard heat polymerized type, autopolymerizing pour type and a heat polymerized rubber graft polymer. These materials were compared and some preliminary results are given. These show that the rubber demonstrated the most satisfactory fatigue life and the pour types were least satisfactory. PMID- 7470554 TI - The polymerization of fibrinogen under the influence of diazomethane modification. AB - To study the effect of the acidic amino acid residues on the physiological polymerization and clot formation of fibrinogen, the fibrinogen system was polymerized by interaction with diazomethane, a specific group reagent which modifies the carboxylic acid residues via the process of methylation. The extent of methylation of fibrinogen by diazomethane was estimated by methoxyl determination on the modified fibrinogen. Under the present experimental conditions, no significant amounts of ammonia were detected as a result of amide hydrolysis concomitant with esterification of the fibrinogen. Chemical methylation of approximately 214 residues resulted in polymerization of the fibrinogen molecule to a product which resembled the physiological clot formation. The application of paper chromatographic techniques identified the modification of other amino acid residues in addition to the methylation of the carboxylic acid groups of glutamic and aspartic acids. These results are interpreted in terms of methylation of carboxylic acid groups in fibrinogen by diazomethane providing a reduction of both negative charge and intermolecular repulsion, thereby enabling the modified fibrinogen molecules to polymerize. PMID- 7470555 TI - Problems and artefacts in the evaluation of polymeric materials for medical uses. AB - The terms 'biocompatibility', 'blood compatibility', and 'thrombogenicity' are often used interchangeably which leads to confusion in the scientific and medical literatures. No single test can reliably predict the 'blood compatibility' of materials, and tests for thrombogenicity are not necessarily indicators of blood compatibility. Although the toxicological screening of biomaterials is important, the absence of adverse effects does not assure biocompatibility. Acute toxicological screenings should be carried in the biological evaluation phases of biomaterials but the results should be interpreted with caution because polymers degrade in the physiological environment by time-dependent mechanisms leading to the formation of potentially toxic products, including carcinogens. Increased efforts are needed to screen potential biomaterials for carcinogenesis and mutagenesis. Biological tests should be conducted with animals whose haematological profile closely resembles that of humans rather than rely on results obtained wity species such as dogs and calves. Accelerated fatigue testing of elastomers must be conducted under condition that closely approximate the properties of the biological environment to avoid catastrophic failures. PMID- 7470556 TI - The material science of calcium phosphate ceramics. AB - Tricalcium, tetracalcium phosphate and hydroxyapatite ceramics exhibit distinct differences in their chemical and structural composition. Only hydroxyapatite ceramic is identical with the original bone mineral. Different preparation methods lead to compact hydroxyapatite ceramic or to porous material with interconnecting macropores as structural equivalents of the spatial structure of cancellous bone. Concerning the behaviour in a biological environment, high crystallinity and large material density result in resistance to dissolution and long lasting stability. Amorphous ultrastructure and porous formation enhance interface activity and bone ingrowth, but also biological degradation of the ceramic implant material. PMID- 7470557 TI - Elastomers for maxillofacial applications. PMID- 7470558 TI - Carbon, a promising material in endoprosthetics. Part 1: the carbon materials and their mechanical properties. AB - Progress in endoprosthetics depends to a large extent on the availability of materials. Carbon is known for its excellent biocompatibility. Carbon materials can also be manufactured with a great variety of properties. The application of carbon materials has been restricted to some special cases like heart valves or tooth roots. Construction of high-loaded endoprosthetic joints has been impossible due to insufficient mechanical strength or manufacturable size of the materials. Three new carbon materials have been developed which seem to offer new possibilities: (1) a high strength isotropic carbon, (2) a silicon carbide/carbon composite (SiC/C) and (3) a carbon fibre reinforced carbon (CFRC). The mechanical properties of these materials were investigated including size effects (length / to thickness d ratio of samples) and manufactured parameters (final heat treatment temperature, fibre volume content and fibre orientation). Flexural strength of the first two materials, which are both isotropic, increases with decreasing l/d ratio, whereas the CFRC behaves contrarily. In the latter case, failure at low l/d ratios is caused by shear failure. The Wohler diagram of all three materials demonstrates limits of fatigue even after only 10(3) to 10(4) cycles. The fatigue strength of the isotropic C and the SiC/C is between 70 and 80% of the static strength, even in Ringer's solution. The Young's modulus of the isotropic C is comparable to that of femoral bone. The modulus of the SiC/C is still appreciably low. It follows that both isotropic materials are applicable for sliding parts like ball and socket. Their tribological properties are described in a following paper. The CFRC finally combines high fatigue strength (approximately 400 MN/m2) with a moderate Young's modulus (approximately 150 X 10(3) MN/m2). These properties indicate application for shafts and pins. PMID- 7470559 TI - Carbon, a promising material in endoprosthetics. part 2: tribological properties. AB - In a previous three types of carbon materials were presented. A basic study of their mechanical properties showed that two of these materials might be applicable for sliding parts of artificial joints such as ball and acetabular cup of a hip joint endoprosthesis. The materials are: (1) a high strength isotropic carbon and (2) a silicon carbide/carbon composite material. The results of relevant tribological investigations are given in this paper. Experimental methods applied consisted of ring-on-disc and ball-in-socket tests. Ring-on-disc tests were performed with specific loads of 10, 20 and 30 MN/m2. In the ball-in socket tests the gap width was 0.075 and 0.1 mm but the load was adjusted in such a way that the compression stress in the contact area was constant at 6.6 MN/m2. The latter was experimentally measured and theoretically calculated with good accordance. Ringer's solution was used as a lubricant. In ring-on-disc tests the ring was made of the SiC/C and the disc of the isotropic carbon. In ball-in socket tests balls of SiC/C and sockets os isotropic carbon were chosen. For comparison the combination CoCrMo alloy or Al2O3 as ball, and HDPE as socket, were tested under corresponding conditions. It is shown, that the special carbon materials can withstand the extreme tribological conditions of high specific load and low sliding velocity. Neither material suffers from fatigue wear. The coefficient of friction is below 0.1 even at a specific load of 30 MN/m2. The wear rate as determined with ring-on-disc test is lower than those of all known and proposed material combinations. The data with the carbon materials combination indicate a penetration of the ball into the acetabular cup of only 1 micrometer per year. PMID- 7470560 TI - Processes human umbilical veins as arterial substitutes--evaluation in canine models. AB - Human umbilical cord vein segments have been used as vessel substitutes for damaged or occluded arteries, as aorto-coronary by-passes and as arterio-venous fistulae for dialysis. The Dardik-Biograft fixed with glutaraldehyde and the Mindich-Bioflow, fixed with ethanol and dialdehyde starch, are commercially available. They were implanted in dogs as replacements for a segment of the abdominal aorta. Post-implantation status was followed by angiography. They were evaluated after removal from sacrificed animals with the aid of scanning electron microscopy and histological techniques. Attention was focused on vessel patency, dimensional stability, integrity of the anastomosis line, lumen wall microstructure, evidence of suture damage and thrombus deposition pattern. Both types of grafts gave functional by-passes for at least until 6 months post implantation. The Dardik-Biograft appeared more prone to thrombus formation near the anastomosis. Sparse cellular development was also noted. The Mindich-Bioflow gave rise to a prosthesis of superior thromboresistance which was more subject to mechanical damage. PMID- 7470561 TI - Materials for internal prostheses: the present position and possible future developments. AB - In this paper we consider solid phase forming as a method of manufacturing acetabular cups. Static and cyclic creep tests have been carried out on Charnley cups and indicate that creep, rather than wear, plays the major role in the penetration of the socket by the femoral ball, at least in the first few years. We have investigated a new type of cross-linked polyethylene and found it to be extremely good from both a wear and creep viewpoint, particularly under high loads and at high sliding velocities. PMID- 7470562 TI - A multi-station machine for the fatigue testing of denture base materials. AB - A multi-station machine has been designed and built to determine the resistance to fatigue failure of denture base materials. The machine has enabled complete S N curves to be plotted for conventional and reinforced denture base materials involving many megacycles of operation. The resistance to fatigue failure of conventional denture base polymers is similar to that of 'Perspex'; that of PMMA reinforced with carbon fibre is of an order of magnitude greater. PMID- 7470563 TI - Electrical stimulation with pt electrodes. IV. Factors influencing Pt dissolution in inorganic saline. AB - Trace analysis has shown that Pt electrodes can suffer appreciable dissolution when used to apply biphasic current pulses of the type used in neural stimulation. The dissolution occurs even under conditions where other irreversible faradaic reactions, e.g., H2O electrolysis, are avoided. In the present study, factors influencing the dissolution of Pt electrodes during biphasic pulsing in neutral inorganic saline have been examined. The findings are consistent with the behaviour of Pt electrodes reported in other inorganic media. In a given test, the quantity of Pt dissolved was found to be a linear function of the aggregate anodic or cathodic charge injected. Therefore, dissolution 'rates' can be conveniently expressed in terms of nanograms of Pt per coulomb injected, e.g., 100 ng C-1. Most of the Pt went into solution as Pt (II) species, so the above rate would correspond approximately to 100 p.p.m. of the anodic charge per pulse. For anodic-first (AF) pulses, charge density was the major factor controlling dissolution, whereas for cathodic-first (CF) pulses, pulse duration had the greater influence. Depending on the polarity (AF or CF), charge density, and duration of the biphasic pulse, the dissolution rate for smooth bead electrodes ranged from 30 to 300 ng C-1. Lower rates were achieved with platinized electrodes but in the absence of organic solutes, it is unlikely that Pt dissolution can be completely suppressed. PMID- 7470564 TI - Electrical stimulation with Pt electrodes. V. The effect of protein on Pt dissolution. AB - The effect of protein on the charge-induced dissolution of Pt stimulation electrodes was studied in an in vitro system. Biphasic pulses (+/- 400 mA, +/- 400 microC cm-2 geom.) were applied to smooth Pt bead electrodes (surface area = 0.03-0.08 cm2) in a buffered saline solution containing 0.075-0.2 mg/ml human serum albumin; pulse solutions were analysed periodically for dissolved Pt by flameless atomic absorption spectrometry. The incorporation of protein in the pulse solution drastically altered the dissolution behaviour of Pt electrodes. Whereas dissolution in organic saline solutions proceeded essentially in a linear fashion, the rate in protein solution decreased with time and asymptotically approached zero. The passage of electric charge was required for the observed inhibition to develop but was not required to maintain it. In contrast, the constant presence of at least 0.15 mg/ml protein in the pulse solution was required for inhibition to develop and continue. Higher concentrations of protein did not enhance the observed inhibition. These findings bear favourably on the prospect for in vivo use of Pt stimulation electrodes: the dissolution process now appears to place no greater restriction on the acceptable pulse parameters than do other irreversible reactions such as H2O electrolysis. PMID- 7470565 TI - Characterization of synthetic apatites for bioceramic implants. AB - Hydroxyapatite has been studied as a substance suitable for surgical substitution of bones and teeth with emphasis on its biocompatibility. The present work tries to identify the characteristics of this material either from the chemico-physical structural point of view, or from the technological one, evaluating the best performance. By X-ray, i.r., thermal, chemical and SEM analyses, the relationship of different phenomena involved in the sintering was evaluated. Technological tests demonstrated the stability and the workability characteristics. In particular, the influence of CO2 was studied, in connection with the most suitable technique for hydroxyapatite (HAP) sintering, considering the eventual aims and requirements for industrial production. PMID- 7470566 TI - Ethylene oxide sterilization: effect of polymer structure and sterilization conditions on residue levels. PMID- 7470567 TI - Kinetics of the aeration of ethylene-oxide sterilized plastics. AB - The aeration of ethylene-oxide (EO) sterilized medical plastic materials is dealth with mathematically, and subsequently the model found is used for calculation of the EO diffusion coefficient in various plastics with the composition and temperature as variables. Absorption and dissolution of EO in various plastic materials are measured at varying concentrations of EO, temperatures and compositions of plastics. The mathematical model is used to evaluate the EO desorption from medical devices under different physico-chemical conditions as well as in vivo. EO analysis methods are evaluated on the basis of the model and are compared with experimental results. PMID- 7470568 TI - Proposed method of intracordal injection and gelation of poly (vinyl alcohol) solution in vocal cords: polymer considerations. AB - Intracordal polymer solution and/or suspension injection has potential in the swelling of paralysed or partially removed vocal cords to the midline laryngeal position. This technique allows the normally functioning cord to vibrate against the swollen one in order to produce sound. Polymeric systems which may possibly be used for the treatment of vocal cords are presented and analysed with special reference to poly(vinyl alcohol). Aqueous poly(vinyl alcohol) solution in concentrations of 3% to 10% (wt/wt) are gelled by glutaraldehyde. The gelation kinetics are followed and the gelation point, final crosslinking density and structure are determined in terms of the initial molar ratio of crosslinking agent to polymer. The mechanical properties of poly(vinyl alcohol) gels are discussed. Finally a proposed technique for the application of these systems for the treatment of certain vocal cord impairments is presented. PMID- 7470569 TI - Quantitative evaluation of water structuring and protein adsorption on the surface of hydrophilic polymers by ESCA. PMID- 7470570 TI - Proposed guideline for the biological testing of orthopaedic implant materials and implants. Prepared by the Working Group on Biomaterials of the German Society of Orthopaedics and Traumatology. PMID- 7470571 TI - Chromatographic isolation and identification of methylated derivatives obtained from modified fibrinogen. AB - Polymerization was accomplished in the fibrinogen system by methylation with diazomethane, thionyl chloride and dimethyl sulphate. The duration and extent of polymerization were dependent on the modifying agent which was applied to the fibrinogen system. When fibrinogen was methylated with a very narrow range group specific methylating agent, like dimethyl sulphate, the polymerization process was accelerated and proceeded with a reduction in the extent of modification of that obtained with the other methylating reagents utilized in these experiments. Chromatographic analysis revealed that diazomethane and thionyl chloride induced both O-methylation and N-methylation, as well as esterification of the carboxylate groups of aspartic and glutamic acid in fibrinogen. However, dimethyl sulphate resulted primarily in esterification accompanied by a small amount of histidine methylation. The proposed mechanism for the polymerization is through the esterification of the aspartic and glutamic acid residues which produces an increased protein-protein interaction resulting in polymer formation. PMID- 7470572 TI - Mechanical properties of some fibre reinforced polymer composites after implantation as fracture fixation plates. AB - Continuous fibre reinforced composite bone plates made from graphite/polysulphone and glass/epoxy laminates were implanted for 16 weeks on osteotomized canine femurs and for 12 months on intact femurs. After sacrifice, the plates were removed and tested in four point bending for stiffness and strength. There were no significant differences in properties between control and implanted plates in the 16 week study. Both the glass/epoxy and the graphite/polysulphone systems showed deterioration after implantation for 12 months. PMID- 7470573 TI - Metal ion release after total hip replacement. AB - The concentration of cobalt (Co), chromium (Cr), molybdenum (Mo), nickel (Ni), iron (Fe) and zinc (Zn) was measured using neutron activation analysis in tissue taken from an 81 year old female at necropsy. The patient had bilateral cobalt chromium molybdenum (Co-Cr-Mo) total hip replacements: one, a metal-on-metal had been in place for 14 years, the other a metal-on-plastic for 5.5 years. Although the metal-on-metal side had become painful, the patient remained active until she died. The measurements indicated that the concentrations of Co and Cr in the lung, kidney, liver and spleen were up to fifty times "Standard Man" values. High values occurred also in the urine and in the hair. The tissue adjacent to the metal-on-metal joint was heavily laden with metal wear debris, whereas that adjacent to the metal-on-plastic joint was relatively uncontaminated. The concentration varied with distance from the implant. Co predominated in the urine, whereas Cr predominated near the implants. The existence of such high levels, especially in the organs, is a possible cause for concern. PMID- 7470574 TI - Polymers containing enzymatically degradable bonds V. Hydrophilic polymers degradable by papain. AB - Copolymers of N-(2-hydroxypropyl)methacrylamide were prepared, in which synthetic polymer chains are joined by crosslinks containing oligopeptidic sequences degradable with papain, with the general structure P-(Gly)n-X-Y-NH-(CH2)6-NH-Y-X (Gly)n-P (P is the polymer chain, n = 1,2; X...Phe, Val, Gly; Y...Lys, Gly, Tyr, Ala, Phe). The relationship between the structure of these polymeric substrates and their degradability with papain was investigated viscometrically. It was shown that -Phe-Lys- was the most suitable -X-Y- sequence. Extension of the oligopeptidic sequence by one amino acid residue causes a pronounced rise in the rate of cleavage of the polymeric substrates. PMID- 7470575 TI - Laminates of poly(2-hydroxyethyl methacrylate) and polybutadiene as potential burn covering. AB - The preparation and the physical characterization of poly(2-hydroxyethyl methacrylate) thermally grafted onto polybutadiene (PHEMA/PB) are reported. The presence of polybutadiene increases the tensile properties of the film and prevents water diffusion through it while it does not affect the oxygen permeability of the PHEMA. The potential use of this material as burn wound cover is suggested. PMID- 7470576 TI - The effect of direct electrical current stimulation on the bone/porous metallic implant interface. AB - Cylindrical porous plugs (6.35 mm dia. 11 mm long, average pore size of 190 micron dia.) made of electrically conductive Co-Cr-Mo surgical alloy powders were implanted in the canine femur. An electrical stimulation device (mercury battery, 1.35 V, connected in series with a 150k omega resistor) was attached to all implants directly. The in vivo current was about 8 microA for the stimulated implants while no current was delivered for the control ones. After predetermined implant periods, tensile test specimens were made to measure the interfacial strength between bone and implants. Some samples were used for histological observations. The present results show that in vivo electrical stimulation substantially increased the strength of the union between porous implants and bone when compared to the controls up to 12 weeks. Histological observations show that the increased strength is mainly due to the increased new bone formation in the pores of implants. It was also observed that the fractional callus volume in the intramedullary canal for the stimulated samples retained more than the controls after reaching maximum at 3 weeks. PMID- 7470577 TI - Polycaproamide degradation in rabbits and in several model media. AB - The nets of polycaproamide removed from rabbits after implantation for times up to 400 days showed a significant change in mass and in molecular weight. The degradation of polycaproamide filaments takes place both on the surface (change of mass) and in the bulk of the polymer implant (change in the molecular weight). The role of phosphates and the enzymes in the polymer degradation are discussed. PMID- 7470578 TI - Alumina as a filler for bone cement: a feasibility study. AB - A composite bone cement of Alcoa A-10 Alumina and very finely ground poly(methyl methacrylate) beads (PMMA) was fabricated. It was tested in an attempt to improve on the conventionally used pure PMMA bone cement. By knowing the densities of the powders and their volumes, the mass of each was calculated for the most efficient packing of PMMA and Al2O3 powders and a 65% PMMA: 35% Al2O3 ratio by weight composition was determined. This was tested, as well as the pure cement so comparisons could be made. Cylinders for the strength tests were also made of silane treated Al2O3. The compositions were tested for compressive and tensile strengths. The pure PMMA, composite and silane treated composite had compressive strengths of 79.64 +/- 13.0, 83.17 +/- 4.8, and 71.52 +/- 8.6 MPa and the tensile strengths were 6.69 +/- 0.6, 5.12 +/- 0.3, and 7.12 +/- 0.5 MPa respectively. Also the 65%-35% PMMA-Al2O3 composite required 64% less monomer for mixing than did the pure cement which is thought to be better for tissue healing. The maximum temperature attained from room temperature was 110 degrees-115 degrees C for both cements. The composite took 6.5 min longer to reach its peak temperature than did the pure cement. The bone cements were implanted for one week in a rabbit and both compositions seemed acceptable by the tissue. PMID- 7470579 TI - Changes of the blood lymphocyte population in cancer patients treated with bestatin, a new immunomodulator. A phase I study. AB - Twenty one patients with advanced metastatic cancer of various types received 30 mg of Bestatin daily per os as a single treatment for several weeks. It was observed that the blood lymphocyte counts remained unchanged but the frequency of SRBC rosette forming cells increased and the frequency of lymphocytes possessing receptors for the Fc-part of IgG became normalized after two weeks of treatment. The frequency of lymphocytes possessing receptors for C'3 was not changed. The natural killer activity of peripheral lymphocytes for K562 and Chang cells increased but the PPD stimulation of the lymphocytes was not altered. A slight, but significant increase of the PHA stimulation of lymphocytes was observed after 4-7 weeks of Bestatin treatment. It is concluded that Bestatin is a nontoxic drug which changes the cellular composition as well as certain immunological functions of human lymphocytes. PMID- 7470580 TI - Detection of lymphocyte stimulation by flow cytometry: differences in cells from patients with and without neoplasia. AB - Fluorescence polarization measurements were performed to detect PHA-stimulation of peripheral lymphocytes in an early state. Single cell fluorescence was measured by flow cytometry using an epi-illumination design to avoid the influence of background fluorescence and multiscattering. By these arrangements the accuracy in the determination of P, the degree of polarization, could be improved over the usual cell suspension measurements. Fluorescence polarization measurements were performed on density specified human lymphocytes after incubation with fluorescein diacetate with and without preincubation with phytohemagglutinin (PHA). A group of healthy donors had an average polarization of P = 0.17 before and P = 0.12 after PHA-stimulation, whereas a group of patients with malignant disease showed an average polarization of P = 0.14 before and after PHA-stimulation. PMID- 7470582 TI - Serial CEA determinations as an aid in postoperative therapy management of patients with early breast cancer. AB - Serial CEA measurements as an aid in routine clinical diagnostic methods was investigated in 69 women with early breast cancer. In 14/69 cases detection of metastatic spread on the basis of elevated pretreatment CEA levels together with clinical aspects led to early adaption of treatment. 27 patients had no metastatic spread (group I) and 28 patients had lymph node metastases (group II). During the follow-up of 55 patients of group I and II, disease progression was signaled by rising CEA values in 10/11 cases with a lead time of up to 8 months before a positive clinical diagnosis was possible. For another 6 patients disease progression has to be expected because of consecutively increasing CEA levels. Patients of group II exhibited a higher frequency of relapses compared to group I patients. Decreasing CEA levels could be correlated in our study with patients who were considered to have had a successful treatment by local radiotherapy and who showed no recurrence during surveillance period along with normal CEA values. About 30% of the patients, however, showed essentially unchanged CEA levels mostly in the normal range. PMID- 7470581 TI - Short and longterm experience with plasmapheresis in connective tissue diseases. AB - Plasmapheresis has been performed in eleven cases with connective tissue diseases, three with systemic lupus erythematosus, three with mixed connective tissue disease and five with rheumatoid arthritis. Plasmapheresis was in all cases instituted as an additional therapy during disease activity which was not under control with conventional modes of therapy. The best results were obtained in systemic lupus erythematosus and rheumatoid arthritis complicated with vasculitis. In mixed connective tissue disease with severe Raynaud's phenomenon, no striking and lasting results were obtained. Plasmapheresis seems to be a valuable adjunct in the management of acute exacerbation of connective tissue disease, especially when it is associated with drugs which can block antibody rebound. PMID- 7470583 TI - Effect of 2,3,7,8-tetrachlorodibenzo-p-dioxin on macrophage and natural killer cell-mediated cytotoxicity in mice. AB - C57Bl/6 J mice (6-8 weeks old) were given single i. p. doses (1, 2, 6 and 30 micrograms/kg) of 2,3,7,8-tetrachloro-dibenzo-p-dioxin (TCDD) and macrophage mediated and natural killer (NK) cell-mediated cytotoxicity was evaluated at different times after treatment. Peritoneal macrophage cytolytic activity was measured as 3H-thymidine release from prelabelled mKSATU5 target cells in a 48 hours assay; macrophage-mediated cytostasis was assessed in terms of inhibitions of 3H-thymidine uptake by SL2 lymphoma cells. Spleen NK activity was measured using 51Cr-labelled YAC-1 lymphoma cells as targets. TCDD did not modify spontaneous macrophage-mediated and NK cell-mediated cytotoxicity per unit number of effector cells nor did it affect the macrophages' capacity to express increased cytolytic and cytostatic activity in the presence of endotoxin. Lower numbers of peritoneal macrophages and splenocytes were recovered from TCDD treated mice. Thus the total numbers of lytic units recovered from animals exposed to TCDD were lower than controls. Impairment of these cellular effector mechanisms, due to cell loss rather than inhibition of function, might play a role in the lowered resistance to bacterial infection of mice given TCDD and in the carcinogenic and cocarcinogenic activity of this chemical. PMID- 7470584 TI - Analysis of visual evoked responses in multiple sclerosis. AB - In Multiple Sclerosis (MS) the modification of the latency of visual evoked responses (VER's) shows sequels of the involvement of anterior optic tracts manifested by a retro-bulbar optic neuritis (RBON). This study was made on 102 patients with MS. The stimulus used the pattern reversal of a checkboard. The influence of the size and squares 8' and 20' chosen as stimulus has been first studied in patients with antecedents of RBON. Using the 8' squares, all eyes with a history of RBON had pathological VER's. When there is no clinical antecedent of RBON, this means of stimulation enabled us to detect sequels of RBON. In "definitive" and "probable" MS, 100% of patients had pathological VER's either for both eyes of a single eyes. In "possible" MS a sequel of RBON was demonstrated in 57% of this population without clinical antecedent of RBON. A correlation between VER's result and the ophthalmological examination (visual acuity, fundoscopic examination and acquired dyschromatopsie) was made. Although VER's are an excellent method for detecting the sequels of RBON in MS. VER's abnormalities have no etiological significance, they are observed in other neurological involvements of anterior optic tracts. These different points are discussed and a physiological interpretation of VER's abnormalities is proposed. PMID- 7470586 TI - Kinetic studies of spin interconversion in ferric mixed-spin derivatives of myoglobin. PMID- 7470585 TI - Kinetics of nucleosome unfolding at low ionic strength. PMID- 7470587 TI - Effect of some organic cosolvents on the reaction of hemoglobin with oxygen. PMID- 7470588 TI - Temperature dependence of the effects of some monohydric alcohols on the oxygen affinity of hemoglobin: determination and analysis of thermodynamic parameters. PMID- 7470589 TI - Characteristics of a volcano field ionization source mass spectrometer operating at 30 kV accelerating voltage. AB - A volcano ionization source has been coupled to an E x B velocity filter type mass spectrometer. The principal advantages of this instrument are its high efficiency, measured at 10(-5) ions per molecule for toluene (m/z 92), and long source lifetimes of over 200 h. The mass spectrometer is designed primarily to obtain mass profiles from multicomponent samples. These profiles are obtained from untreated urine and blood serum (1 microliter and 0.25 microliter samples), then measured between m/z 50 and 400, and are finally analyzed by computer. PMID- 7470590 TI - Simultaneous determination of bestatin and p-hydroxybestatin, a major metabolite, in human serum by gas chromatography mass spectrometry. Utilization of quantitative selected ion monitoring and deuterium labelled internal standard. AB - A specific and sensitive method for the simultaneous determination of bestatin and its major metabolite, p-hydroxybestatin in human serum has been developed by selected ion monitoring using the corresponding deuterated internal standards. The N-trifluoroacetamide-O-trimethylsilyl either derivatives of the trideuteromethyl esters of bestatin and p-hydroxybestatin were suitable for quantitation by selected ion monitoring. The determination limit of these compounds by the present method is 1 ng ml-1 in human serum. PMID- 7470591 TI - Effects of liver disease on urinary excretion of methadone and metabolites in maintenance patients: quantitation by direct probe chemical ionization mass spectrometry. AB - This study was performed to define the amounts of methadone and metabolites excreted in urine in otherwise healthy maintenance patients, and to determine whether the metabolism and elimination of methadone, as assessed by analyses of urines, is altered in patients with liver disease. A method was developed for the simultaneous quantitation of methadone and six of its major and minor metabolites using chemical ionization mass spectrometry with direct probe introduction to increase sensitivity for analyses of the minor metabolites. Analyses of urine from unmedicated volunteers showed that the interferences at the mass range of interest (264-326) were usually small and therefore would not introduce significant error into analysis. Nineteen patients well-stabilized in chronic long-term methadone treatment were studied, five otherwise healthy males and fourteen patients with chronic liver disease (nine males and four females). Twenty-four hour urine collections were made and analyzed following extraction procedures. The concentrations of methadone and the major pyrrolidine metabolite exceeded 1 microgram ml-1 in all cases; the concentration (listed in descending order) of pyrrolidone, pyrroline, hydroxymethadone, hydroxypyrroline, methadol and hydroxypyrrolidine were all less than 1 microgram ml-1. The total 24 hour urinary excretion of methadone and its metabolites was 48.3% (+/- 1.71 SEM) in otherwise healthy patients but was significantly lower, 32.6% (+/- 3.19 SEM) in patients with liver disease (p less than 0.05). The total 24 hour excretion of the pyrrolidone metabolite, the end product of two pathways of methadone metabolism, was also significantly reduced in patients with liver disease (p less than 0.05). Females with liver disease had significantly higher ratios of pyrrolidine to methadone than did males with liver disease (p less than 0.05). PMID- 7470592 TI - An improved method for determination of plasma norepinephrine: isolation by boric acid gel and assay by selected ion monitoring. AB - An accurate and time-saving method for the determination of norepinephrine in human plasma is described. The whole device focused on an improvement in the pretreatment. One ml of human plasma was applied to a boric acid gel column without deproteinization and pH adjustment, and norepinephrine was eluted with 2 ml of a 1.3 M acetic acid in methanol solution. The pentafluoropropionic derivative of norepinephrine was analysed by gas chromatography mass spectrometry using deuterated norepinephrine as an internal standard. The level of plasma norepinephrine in healthy subjects at recumbency was 297 +/- 95 pg ml-1 (mean +/- SD). The lower limit of sensitivity was 25 pg ml-1 of norepinephrine in plasma. PMID- 7470593 TI - [Role of hypothalamic monoaminergic structures in regulating sympatho-adrenal system function]. AB - The effect of pharmacological destruction of hypothalamic monoaminergic terminals on the basal and cold stress-induced secretion of neurohormones of the sympathoadrenal system (SAS) in rats was studied. The data obtained on biochemical and histochemical analysis of the hypothalamus (dopamine, noradrenaline and serotonin level, the picture of catecholaminergic terminal fluorescence) were compared with adrenaline and noradrenaline content in blood. It is assumed that function of the SAS is inhibited by hypothalamic noradrenaline and stimulated by dopamine and serotonin under cold stress. PMID- 7470594 TI - [Role of contact with a support in the tumbling response of white rats during free fall]. AB - The tumbling and landing reactions of white rats were studied by cinematography during their free fall with eyes open. If the rats let the support go, they turned over and made a soft landing on their paws, no matter what their initial position was. However, when the animals fell down together with the support, the tumbling reaction was inhibited entirely and they landed as they were in the initial position, i.e. on the back, head, tail, etc. If during free fall the animals crumpled the support or if they touched a fixed object with their limbs or tail, they turned over and landed on their paws. PMID- 7470595 TI - [Reciprocal connections of respiratory center nuclei]. AB - The reactions of 181 respiratory neurons of solitary and ambiguous nuclei were studied on 64 cats in the course of stimulation of inspiratory and expiratory zones of gigantocellular nuclei. A study was also made of the reactions of 224 reticular and 9 respiratory neurons of these zones during stimulation of solitary and ambiguous nuclei. Various reactions were recorded in response to the stimulation. Therefore the nuclei of the respiratory center possess various information. In the course of stimulation of the gigantocellular nucleus there predominated an increase in the neuronal electrical activity. On the contrary, inhibition of respiratory and reticular neurons of the gigantocellular nucleus was predominant in the course of stimulation of solitary and ambiguus nuclei. It is suggested that the interactions described underlie the formation of circular rhythms of respiratory center stimulation. PMID- 7470596 TI - [Dynamics of restoration of the baseline pH level in the stomach following active gastric secretion induced by a food stimulus]. AB - Intragastric pH-graphy was used in experiments on fistulous dogs to study the time course of pH basal level recovery in the stomach after active gastric secretion induced by food irritant. The time-dependent characteristics of pH waves is described in an attempt to ascertain their genesis. The notion "gastric cycle" is validated. It is recommended that in the course of determination of gastric secretory function, the features of pH basal level recovery, including acid neutralization and cessation of acid formation should be taken into consideration along with other parameters. PMID- 7470597 TI - [Redistribution of fluid phases of the brain during an increase in blood osmolarity]. AB - Intravenous injection of polyethylene glycol 400 to rats (1.5 mmole/100 g body weight) increases blood osmolarity, the total water content in the brain hemispheres exhibiting insignificant changes. Cellular dehydration is accompanied by an augmentation of the volume of extracellular fluid in the intact skull cavity. The increased blood content of the brain is pointed to be a 24% increase in the iron level in its tissues. Skull trepanation enhances the dehydrating effect of polyethylene glycol 400 but does not affect the blood content of the brain. PMID- 7470598 TI - [Inhibition of lipid peroxidation during emotional-painful stress by ionol and gamma-hydroxybutyric acid]. AB - UV spectroscopy, polarography and fluorescence of Schiff's bases were used to study the time course of emotional pain stress-induced accumulation of the primary and secondary products of lipoperoxidation (LPO) in different tissues of Wistar rats and to appraise the effect of gamma-hydroxybutyric acid (GHBA) and 4 methyl-2,6-ditretbutylphenol (ionol) on LPO induction. LPO activation was shown to be retained after emotional pain stress for a long enough period of time. This activation may last from 2 to 5 days depending on the organ. Pretreatment with GHBA or ionol completely reverses LPO activation. PMID- 7470599 TI - [Dynamics of the concentration of aldosterone, cortisol, and its precursors in the plasma of M. rhesus during long-term restriction of motor activity]. AB - A significant increase in the content of cortisol and its precursors was detected in blood plasma from rhesus monkeys during the first six hours of the motor activity restriction. In the three days following, the peak concentrations of cortisol were accompanied by a fall of 17-hydroxylated precursors to the basal and lower levels. The content of cortisol attained the basal level by the end of the first week of fixation. The levels of cortisol, 11-deoxycortisol, 17 hydroxyprogesterone and 17-hydroxypregnenolone on the 17th day of hypokinesia were 77, 62, 39 and 15%, respectively, as compared with the pre-experimental period. The maximum rise of the aldosterone level in monkey plasma was seen on the 2--3d day followed by a gradual decrease in its concentrations over the entire experimental period. It is suggested that the reduced blood content of steroids during the restriction of motor activity is related to the adaptation of pituitary-adrenal system function to the decreased environmental requirements. PMID- 7470600 TI - [Age-specific differences in the process of adrenal cortex cell membrane repolarization]. AB - The differences in temperature dependence of the membrane repolarization process (RP) in the cells of zona fasciculata of the adrenal cortex within the range of 7 -17 degrees C following preliminary cooling of the adrenal glands to 0 degrees C were investigated in experiments on the isolated adrenal glands of rats aged 5 and 28--29 months. The temperature coefficient (Q10) of RP was much higher in old (2.732 +/- 0.191) than in young rats (1.481 +/- 0.051). Ionol, an inhibitor of free-radical reactions (FRR), did not change Q10 of RP in young animals but decreased it significantly in old ones down to the level of young rats (1.467 +/- 0.028). The total x-raying of young rats (700 Gy) led to an increase in Q10 of RP to 2.515 +/- 0.099 in the first hours after exposure to x-rays. This parameter returned to normal after 48 hours. Age-associated reconstruction of the electrogenic active transport mechanisms is supposed to lie in the appearance (or increased contribution) of FRR in the total balance of reactions determining the active transmembrane transport processes. PMID- 7470601 TI - [Fatty acid hydroperoxides, fluorescent products, and tocopherol concentration in the tissues of rabbits exposed to cold]. PMID- 7470602 TI - [Effect of prednisolone on 3H-catecholamine synthesis in the adrenals of physically fatigued rats]. AB - The effect of prednisolone on the synthesis of 3H-catecholamines by the adrenals of control and rats swimming for 8 hours was studied in experiments in vitro with the use of 3H-thyroxine as precursor. It was shown that in swimming rats, the synthesis of 3H-noradrenaline and 3H-adrenaline is inhibited by 25%, while the synthesis of 3H-dopamine remains unchanged. Addition of prednisolone to the incubation medium does not affect the rate of 3H-catecholamine synthesis by the adrenals of intact rats. Addition of prednisolone to the adrenals of swimming rats leads to the stimulation of the synthesis of 3H-noradrenaline and 3H adrenaline by 72%, while the synthesis of 3H-dopamine gets stimulated by 56% relative to the synthesis rate of 3H-catecholamines during swimming. In the latter case the rate of 3H-catecholamine synthesis significantly exceeds the control level. PMID- 7470603 TI - [Quantitative determination of circulating antivascular antibodies in atherosclerosis by the autocomplement consumption method]. PMID- 7470604 TI - [Effect of growth hormone on polynucleotide phosphorylase activity in a primary monolayer culture of rat hepatocytes]. AB - Growth hormone (1 microgram/ml) inoculated into the primary culture of rat hepatocytes and incubated for 3 hours at 37 degrees C decreases 2-fold the activity of polynucleotide phosphorylase (PNPase). In the culture of hepatocytes from hypophysectomized rats, the basal activity of PNPase is significantly higher as compared with that in the culture of hepatocytes from intact animals. The most pronounced inhibitory effect of growth hormone on PNPase was seen in the culture of hepatocytes from hypophysectomized animals. PMID- 7470605 TI - [Anticonvulsive effect of the antioxidant ionol]. AB - The influence of the antioxidant ionol on the intensity of epileptic activity in rats has been studied. The epileptic activity was induced by penicillin application to the sensorimotor cortex. It was established that preliminary injection to rats of ionol abolished the effect of lipoperoxidation in the focus of hyperactivity and decreased by a factor of 10(1) the number of seizures recorded on the electrocorticogram in the course of the focus existence. PMID- 7470606 TI - [Effect of odiphalin and difril on catecholamine concentration in adrenergic nerve fibers of different rat organs]. AB - The histochemical method of Axelsson coupled with photometry was used to study variations in the content of biogenic amines in different rat organs (myocardium, vas deferens, mesenteric vessels, iris, atrial and ventricular heart valve) under the effect of odiphalin and difril. The results obtained indicate that the organs with direct, indirect and mixed innervation differ in the pattern of response to the pharmacological agents administered. Venous vessels proved to be the most sensitive to the action of odiphalin and difril. The data obtained warrant a conclusion about the heterogeneity of adrenergic nerves in organs and tissues. PMID- 7470607 TI - [Effect of etimizol on hematocellular barrier permeability in hepatitis induced by carbon tetrachloride poisoning]. AB - It was established in experiments on 47 rats that administration of carbon tetrachloride entails the increased permeability of the hematocellular barrier of the heart, lungs, skeletal muscles, adrenals, liver and kidneys. Administration of ethimisole promotes normalization of the permeability of the test organs. PMID- 7470608 TI - [C1q component of complement and collagen reaction with nucleic acids and polyanions]. AB - Interaction of acid-soluble collagen and C1a component of complement with nucleic acids (NA) is defined by their polyanionic properties. Occurrence of the exocyclic amino group in the structure of nitrous bases from which polynucleotides are made up inhibits their interaction with the proteins are made up inhibits their interaction with the proteins even when this group is within the double helix. This takes place during the formation of the hybrid molecule poly I-poly C. It is assumed that the collagen-like part of C1q greatly contributes to its interaction with nucleic acids. PMID- 7470609 TI - [Prevention of experimental allergic encephalomyelitis in guinea pigs by treatment with magnetophores]. AB - Subcutaneous application of magnetophores with a field intensity of 210 ersted prevented the development of experimental allergic encephalomyelitis (EAE) in guinea-pigs. The magnetophores appeared efficacious both at the beginning of the induction period and in the second half of the latent period, i.e. within the first two weeks after injection of encephalitogenic suspension. Application of magnetophores after the appearance of the neurological signs of EAE was ineffective. Magnetophores did not affect cellular or humoral response in animals. It may be assumed that the effect of magnetophores is related to the increased resistance of nerve cells to the immunopathological action of the immune response factors, a topic for further studies. PMID- 7470610 TI - [Effect of nitrosoethylurea on sarcomagenesis induced by a foreign body]. AB - Subcutaneous administration to rats of an intact cellophane plate gave rise to sarcomas in 60% of cases, the latent period being 16.5 month. Meanwhile administration of minced cellophane did not produce tumors. Combination of the plate subcutaneously and nitrosoethylurea (NEU) intraperitoneally increased the frequency of sarcomas to 73.3%, the latent period being 13.7 month. Sarcomas emerged in 33.3% of cases at the site of administering minced cellophane coupled with intraperitoneal injection of NEU, the latent period amounting to 13.2 month. The morphological appearance of sarcomas that occurred with the use of NEU differed from tht resultant from administering the cellophane plate alone. Adsorption of NEU on the cellophane is suggested. PMID- 7470611 TI - [Estradiol stimulation of thyrotropic hormone elaboration in cultures of adenohypophysis obtained from intact and ovariectomized rats]. AB - The secretion of thyrotropin by pituitary cells in culture from ovariectomized rats treated with estradiol dipropionate was after 3-4 days higher than in cultures from intact and ovariectomized rats. Moreover, 17 beta-estradiol directly stimulated the secretion of thyrotropin by adeno-hypophyseal cells of intact rats in a monolayer culture. PMID- 7470612 TI - [Features of porphobilinogen synthesis from delta-aminolevulinic acid in the tissues of the internal organs of rats]. AB - The biosynthesis of porphobilinogen by homogenates of the lungs, heart, liver, kidneys, spleen, pancreas and small intestinal tissues was studied in 77 intact and in 68 rats with marked lead poisoning. The activity of ALA dehydratase in the lungs was decreased by 40.3%, in the kidneys by 40%, in the heart by 38.2% and was normal in homogenates of the liver, pancreas and small intestine. In spleen homogenates, the activity was increased by 26.3%. Potential mechanisms of different ALA dehydratase reactions in organs tissues of rats with lead poisoning are discussed. PMID- 7470613 TI - [Subcellular localization of 45Ca2+ during absorption by small intestinal epithelium in rats]. AB - Absorption of 45Ca2+ administered in to the lumen of the young rat small intestine at a concentration of 0.25 mM has been examined by electron microscopy coupled with autoradiography. 45Ca2+ has been shown to be transported through an enterocyte in the sequestered form within the membrane vesicles that detach at the base of microvilli, opening at the lateral membranes of the enterocyte, within Ca2+ being released into the intercellular space. PMID- 7470614 TI - [Lymphoid cell ultrastructure in experimental tuberculous granuloma]. AB - There are two groups of activated cells differing both in ultrastructure and manifestations of functional activity in lymphoid populations participating in the formation of tuberculous granuloma. In group 1 lymphoid cells, the development of intracellular structure is associated with an intense synthetic and secretory activity. Group 2 cells are mainly characterized by absorption function realized by means of active pinocytosis. The heterogeneity of subcellular organization of lymphoid cells in experimental tuberculosis reflects differences in their functional specialization. Study of tuberculous granuloma lymphocytes by scanning electron microscopy does not allow the lymphocytes to be identified as T and B cells without immunologic marker reactions. PMID- 7470615 TI - [Dynamics of morphologic changes during treatment of experimental acute pancreatitis with 5-fluorouracil and intralipid]. AB - In experimental pancreatitis induced by the modified Agai technique, the combined administration of 5-fluorouracil and intralipid produced an appreciable therapeutic effect, preventing the development of acinus necrosis and suppurative melting of the gland tissue. It is suggested that the effect of the pharmacological combination is consequent on the inhibitory action of 5 fluorouracil on DNA and RNA synthesis by exocrine cells of the pancreas and on the stabilizing action on the cellular and capillary membranes of unsaturated fatty acids constituting intralipid. PMID- 7470616 TI - Fibrinogen receptor exposure and aggregation of human blood platelets produced by ADP and chilling. PMID- 7470617 TI - Decreased deformability of erythrocytes from smokers. AB - The deformability of erythrocytes from smoking and non-smoking human subjects was examined by filtration through 3-mu pores and capillary viscometry of cell suspensions. In both cases, small but significant differences were found between the two groups that are consistent with a reduction in the surface area-to-volume ratio and/or a diminished membrane flexibility in erythrocyte from smokers. Additional evidence suggests that these findings represent a chronic rather than an acute effect of smoking on erythrocyte deformability. PMID- 7470618 TI - Negative charge distribution and density on the surface of oxygenated normal and sickle red cells. AB - Negative charges on the external surface of red cells were visualized by colloidal iron hydroxide labelling of 50% of the membrane area after osmotic hemolysis and glutaraldehyde fixation. Counts were made over randomly selected areas on electron micrographs at 350,000 x magnification. Statistical analyses showed that at the 95% level of confidence there was no significant difference between oxygenated normal (AA) and sickle (SS) cells in either the distribution or the density of negative charges. PMID- 7470619 TI - Characteristics of the synergistic actions of platelet agonists. AB - Platelets are activated by many different agonists that act synergistically. Since there is a characteristic pattern of responses to each agonist, and since there is a clear distinction between weak and strong agonists, understanding the nature of the synergism and its physiologic significance requires characterization of the pattern of responses to the synergistic action of the various agonists. Shape change, aggregation, and secretion of ATP by human platelets in citrated plasma were analyzed after activation by ADP, epinephrine, arachidonic acid, gamma-thrombin, or collagen, either singly or in pairs. The patterns of responses were characteristic of the agonist in higher concentration relative to its threshold concentration; if neither was clearly higher, the pattern of responses was intermediate between the responses characteristic of each agonist. No combination of weak agonists had the characteristics of a strong agonist. These results help define the extent to which platelet responses can be attributed to the synergistic actions of weak agonists. PMID- 7470620 TI - Hb Toyoake: beta 142 (H20) Ala replaced by Pro. A new unstable hemoglobin with high oxygen affinity. AB - A new unstable hemoglobin with high oxygen affinity, Hb Toyoake: beta 142 (H20) Ala replaced by Pro, was found in Japanese male with a normal blood hemoglobin level, shortened red cell survival, and increased plasma erythropoietin. Hemoglobin studies showed heat and isopropanol instability, and an increased tendency to heme loss and to subunit dissociation. Electrophoresis of whole hemolysate showed inconstant abnormal bands with reduced mobilities due to progressive heme loss during the in vitro procedure. Isolated Hb Toyoake with normal heme content migrated slightly faster than HbA. Oxygen affinity of red cells was elevated with P50 of 17.0 mm Hg at pH 7.4 and 37 degrees C (normal 25.0 mm Hg). Studies on hemolysate implied that Hb Toyoake had an almost normal Bohr effect, a diminished cooperativity, and a reduced response to inositol hexaphosphate. These multiple abnormalities are associated with a substitution of Pro for beta 142 Ala, resulting in disruption of the H-helix and the adjacent C terminal portion of beta chain, which contain residues crucial for normal oxygen binding. PMID- 7470621 TI - Hemoglobin Petah Tikva (alpha 110 ala replaced by asp): a new unstable variant with alpha-thalassemia-like expression. PMID- 7470622 TI - Acute nonlymphocytic leukemia, preleukemia, and acute myeloproliferative syndrome secondary to treatment of other malignant diseases. Clinical and cytogenetic characteristics and results of in vitro culture of bone marrow and HLA typing. PMID- 7470623 TI - Procoagulant activity of sarcoma sublines with different metastatic potential. AB - It has been suggested that cancer cell procoagulant activity influences metastasis formation by promoting fibrin deposition around tumors. We have investigated the procoagulant activity of various tumor cell sublines with different metastatic capacity derived from metastatic nodules of a murine fibrosarcoma. All the cells tested possessed a marked thromboplastin-like activity; they were, however, heterogeneous as regards the degree of procoagulant activity; the two cell lines with virtually no metastatic capacity showed 6--8 times higher procoagulant activity than the cells from the parent line; in contrast, the procoagulant activity of the two sublines with higher metastatic capacity did not differ significantly from that of the parent line. These findings support the hypothesis that fibrin is part of a defence reaction against cancer cell invasiveness. PMID- 7470624 TI - Density subpopulations of rabbit platelets: size, protein, and sialic acid content, and specific radioactivity changes following labeling with 35S-sulfate in vivo. PMID- 7470625 TI - The role of the nuclear protein matrix during development of rabbit granulocytes. AB - A proteinaceous nuclear substructure (nuclear protein matrix or nuclear pore complex-lamina) has been described in a number of cells and may be a universal feature of cell nuclei. We have investigated the nuclear protein matrix (NPM) in the rabbit blood granulocyte and its precursor cells to determine (A) whether the NPM composition is similar to other cells that have been studied and (B) to determine whether the dramatic morphological changes that the granulocyte nucleus undergoes during cell maturation are related to changes in the composition or structure of the NPM. NPM preparations from rabbit granulocytes were similar but not identical to those found in HeLa cells or rat liver nuclei. The NPM structure of more mature cells retained more DNA during the isolation procedures than did immature cell NPMs, and the DNA was less accessible to DNAse in the mature cell nuclei. Scanning and transmission electron microscopy revealed a continuous outer covering of the NPM preparation and a lattice-like internal structure. Recognizable nuclear form persisted although the preparations represented less than 20% of the original nuclear protein. NPM preparations from mature cells were similar in overall dimensions and internal structure to NPM from immature cells, suggesting reexpansion during isolation of a previously compacted NPM structure in segmented neutrophils. NPM proteins are synthesized primarily in early stages of cell development. The NPM appears to play a major, but passive, structural role in the nuclear changes observed during maturation of granulocytes. PMID- 7470626 TI - Quantitative measurement of C3 activation at polymer surfaces. PMID- 7470627 TI - Human megakaryocyte stimulation of proliferation of bone marrow fibroblasts. AB - Human marrow cells were processed sequentially by density centrifugation and by velocity sedimentation in serum-free Percoll gradients in order to purify megakaryocytes and to determine if these cells are the source of the growth factor derived from platelets. Cell homogenates were made from the resulting fractions and tested for growth-promoting activity(ies) in 3T3 cells and in well characterized human marrow fibroblasts. Growth was evaluated by 3H-TdR incorporation and changes in DNA cell content, as measured by flow microfluorometry. The highest mitogenic activity was derived from homogenates of low density (less than 1.050 g/cu cm), rapidly sedimenting cells. This fraction contained the highest percentage of megakaryocytes. The assessment of growth promoting activity(ies) derived from various megakaryocyte-enriched marrow cell homogenates containing different proportions of megakaryocytes demonstrated a positive correlation between the number of megakaryocytes and their stimulatory capacity as determined by 3H-TdR uptake. The growth-promoting activities elicited from homogenates of platelets and marrow fractions enriched for megakaryocytes were similar. The dose--response curves for both were parallel, and they were both temperature resistant and trypsin sensitive. These findings implicate megakaryocytes as a source of the growth factor derived from platelets and suggest that megakaryocytes may play a role in the pathogenesis of the marrow fibrosis observed in myeloproliferative disorders by stimulating fibroblast proliferation and collagen secretion. PMID- 7470628 TI - Imbalanced globin chain synthesis in cultured erythroid progenitor cells from thalassemic bone marrow and peripheral blood. PMID- 7470629 TI - Cell sorting: what to sort and why? PMID- 7470630 TI - Lectin receptors on human blood and bone marrow cells and their use in cell separation. AB - A series of fluorescein-conjugated lectins (Sophora japonica agglutinin, Helix pomatia agglutinin, peanut agglutinin, the erythroagglutinin from Phaseolus vulgaris, pokeweed mitogen, wheat germ agglutinin and the fucose-binding lectin from Lotus tetragonolobus) were analyzed for their binding to human peripheral blood cells with a fluorescence-activated cell sorter. Most of the lectins showed increasing cell binding in the order erythrocytes less than lymphocytes less than monocytes less than neutrophils and the degree of fluorescence was not related to the major blood groups. The fucose-binding lectin (FBP) was the main exception in that it appeared to bind only to blood neutrophils. From an analysis of the binding of this lectin to human bone marrow cells it was found that the degree of binding within the granulocytic series increased with progressive differentiation. Marrow monocytes and nucleated erythroid cells bound to FBP in contrast to monocytes and non-nucleated red cells in the peripheral blood which showed negligible binding. Lymphocytes both in the marrow and blood displayed negligible binding of FBP. These properties allowed an enrichment of haemopoietic progenitor (colony-forming) cells from human marrow cell suspensions and a depletion of colony-inhibiting cells, when these were present, by selection of cells with the appropriate fluorescence intensity. PMID- 7470631 TI - Sources of haemopoietic stem cell proliferation: stimulators and inhibitors. AB - Based on earlier findings that haemopoietic tissue contains extractable factors which are capable of specifically inhibiting or stimulating the movement of CFU-S into DNA synthesis, a series of preliminary experiments has now been carried out to investigate their cellular source(s), their activity in vivo, and their applicability to human problems. In vivo treatment of mice, in which femoral CFU S are proliferating rapidly, with the inhibitory factor reduces the proportion of CFU-S in DNA synthesis to non-significant proportions. In addition, the inhibitor is capable of reducing the number of CFU-S induced to enter S following treatment with hydroxyurea, thus protecting CFU-S from the lethal effects of S-phase cytotoxic agents. Removal of specific types of marrow cells shows that both inhibitor and stimulator are adherent, phagocytic and, in the case of inhibitor, Thy--1-. These results suggest that the producer cells probably reside somewhere in the heterogeneous macrophage complex though their different densities suggest they are probably different cell types. Fresh human bone marrow is found to contain a very similar inhibitor and long-term cultures are also found to produce it continuously. The isolation of the producer cells may thus contribute to the understanding of normal physiological stem cell regulation and, by in vivo application, its eventual manipulation and protection. PMID- 7470632 TI - Motion, deformation, and interaction of blood cells and plasma during flow through narrow capillary tubes. AB - The microrheological characteristics of human blood cell suspensions flowing through very narrow glass capillaries (I.D. 5-12 microns) have been investigated using a traveling capillary method. In addition, red cell shape during flow was studied after cell fixation during capillary passage. The observations and measurements indicate the following: 1. The deformed shape of human red cells is not axisymmetrical and develops from an edge-on-oriented biconcave disc shape. 2. Red cells tend to travel in groups rather than equally spaced, presumably due to interindividual differences in size, shape, and deformability. 3. Red cell rotation in 8-12 microns capillaries occurs only if the flow forces are too small to induce cell deformation; during rotation of biconcave discs a substantial fraction of orbiting times is spent in an edge-on orientation; rotation is more frequently observed if the deformability of the erythrocytes is reduced. 4. Hematocrit-dependent transition from single-file to multi-file ('zipper') flow is observed in tubes whose inner diameter exceeds approximately 6 microns. 5. The flow pattern in the larger capillaries (8-12 microns) is characterized by significant variation of both radial and axial velocity of individual cells, as well as by concomitant changes of cell shape. However, cell-to-cell interactions and changes of cell orientation become progressively less significant in even smaller capillary tubes. 6. The bolus motion of intercellular plasma can be demonstrated by using microspheres or single platelets flowing between red cells as indicators. 7. Single platelets flowing between erythrocytes exhibit considerable variation in axial velocity in addition to rapid rotation. PMID- 7470633 TI - Receptors for 5-hydroxytryptamine in rabbit isolated ear artery and aorta. AB - The effects of N,N'-bis-(O-methoxybenzylaminohexyl)-cystamine tetrahydrochloride (BHC), 2-brom-d-lysergic acid diethylamide (BOL) and prazosin on 5 hydroxytryptamine (5-HT) dose-response curves in rabbit ear artery and thoracic aorta were determined. BHC, an irreversible alpha-adrenoceptor antagonist, and prazosin had no substantial effect while BOL shifted the 5-HT dose-response curve to the right in aorta. BHC shifted the curve to the right and reduced maximal response to 5-HT in ear artery. BOL shifted the curve to the right only above 10( 6) M 5-HT in control, but at all concentrations studied in BHC-pretreated ear arteries. In vitro denervation of the ear artery with 6-hydroxydopamine did not significantly alter the 5-HT dose-response curve, nor the effect of BHC on that curve. On the other hand, desipramine decreased the contractile response to high concentrations of 5-HT in non-denervated ear arteries. Prazosin increased the 5 HT threshold and slope of the curve in the ED50 region, but had no inhibitory effect on contractile responses above 3 x 10(-7) M 5-HT or on maximal response. It is concluded that 5-HT acts exclusively on 5-HT receptors in aorta, but on both alpha-adrenergic and 5-HT receptors in ear artery. 5-HT also possesses a small indirect sympathomimetic action at high concentrations in the ear artery. Prazosin has no effect on 5-HT receptors in either vessel and blocks the alpha adrenoceptor stimulation by 5-HT in ear artery. PMID- 7470634 TI - Inhibitors of mitochondrial Ca++ uptake dissociate potassium-induced tension responses from increased 45Ca retention in rabbit aortic smooth muscle. AB - Effects of antimycin A, oligomycin and KCN on tension responses to high K+ or NE and on related Ca++ movements were investigated in rabbit aortic smooth muscle. Selected concentrations of antimycin A (10(-6) g/ml), oligomycin (10(-6) g/ml) and KCN (1 mM) had little effect on the K+-induced contractile response even though all three agents abolished the increase in La+++-resistant low affinity Ca++ uptake elicited with high K+. Though the mitochondrial inhibitors had varied effects on the NE-induced contractile response, they had no effect on the NE induced decrease in La+++-resistant high affinity Ca++. Oxygen consumption of the muscle tissue was decreased by antimycin A or oligomycin. These results suggest that the increase in low affinity Ca++ retention elicited with high K+ represents Ca++ uptake by mitochondria. Thus, the correlation between K+-induced uptake of Ca++ and an increased retention of La+++-resistant low affinity Ca++ after exposure to K+ are sequential events that can be associated by inhibitors of mitochondrial Ca++ uptake. PMID- 7470635 TI - Effect of transmembrane pH gradient changes on potassium-induced relaxation in vascular smooth muscle. AB - In bicarbonate-buffered solution the Ba++-induced contraction of rat aorta was relaxed by an elevated extracellular K+ concentration. When the pH of the bicarbonate-buffered solution was lowered or the buffer system of the solution changed from bicarbonate to Tris or HEPES, the K+-induced relaxation was strongly inhibited. Raising the pH of the Tris-buffered solution restored the effect of K+. Addition of NH4+, which is known to cause a transient increase in intracellular pH, transiently inhibited and subsequently augmented the K+-induced vasodilation in all solutions. It is suggested that the K+-induced vasodilation results from membrane hyperpolarization following enhanced Na+, K+-ATPase activity and that this K+-induced vasodilation is affected by changes in the transmembrane pH gradient. PMID- 7470636 TI - Exercise hyperemia in the absence of a tissue PO2 decrease. AB - The relationship between contraction-induced arteriolar vasodilation, estimated blood flow (BF), and tissue oxygen tension (PtO2) in skeletal muscle of young rats was investigated. Arteriolar diameters, BF, and PtO2 were measured during cremaster muscle stimulation at 2, 4, and 8 Hz. At a suffusate PO2 (PsO2) of 5-10 mm Hg, the caliber of second-order arterioles (2A, greater than 30 micrometer diameter) did not significantly increase after 8 Hz stimulation. In contrast, with the same PsO2, third-order arterioles (3A, less than 30 micrometer diameter) significantly dilated to 127 +/- 9 (SE), 135 +/- 12, and 160 +/- 13% of control, respectively at 2, 4, and 8 Hz stimulation. Third-order arteriolar BFs at the end of the stimulation periods were: 119 +/- 18, 176 +/- 14, and 258 +/- 43% of control, respectively. PtO2 remained essentially constant during and following 4 and 8 Hz stimulation if the systemic arterial blood pressure exceeded 80 mm Hg. When PsO2 was increased to 65-70 mm Hg, neither 3A diameters nor PtO2 during 8 Hz stimulation differed from paired comparisons of the response at low PsO2. These data indicate that dilation of small arterioles during increased muscle metabolic activity can occur without a mandatory change in PtO2. This circumstance is unaffected by a substantially increased supply of oxygen to the tissue. PMID- 7470637 TI - Cellophane perinephritic hypertension in the dog: some biochemical, hemodynamic and pathological characteristics. AB - Perinephritic hypertension was induced in mongrel dogs by cellophane encapsulation of one kidney and simultaneous contralateral nephrectomy. Systolic blood pressure began to rise before diastolic blood pressure and both reached maximal values 3 weeks after surgery, their levels becoming stable thereafter up to the end of the study (20 weeks). Cardiac contractility, measured by the index dp/dt/P40 (rate of rise of isovolumic left intraventricular pressure at a developed left ventricular pressure of 40 mm Hg) was not altered during the evolution of hypertension; heart weight and left ventricular thickness were increased. Plasma renin activity markedly decreased in the chronic phase and plasma urea and creatinine levels rised initially in a moderate but significant way, thereafter remaining stable. The structure of the wrapped kidney remained normal 20 weeks after surgery, when microscopic arterial lesions were seen in other organs. The comparison of the evolution of this hypertensive model with other more commonly used renal experimental hypertension types allows us to conclude that cellophane perinephritic hypertension in the dog must be considered as an individual entity. PMID- 7470638 TI - Contractility and the length-tension relation of the dog anterior tibial artery. AB - Active force (S) and rate of change of force (S) throughout the time of isometric contraction in electrically stimulated, circumferentially cut segments of dog anterior tibial artery were analyzed for dynamic indices of the contractile state. Previous works have determined indices that change with temperature but not with length (L) or calcium (Ca) and were designated indices of crossbridge dynamics. They assumed that changes in L and Ca change only the number of crossbridges between contractile filaments and temperature affects only the dynamics of individual crossbridges. We have shown: (1) specific ratios of S/S to be independent of L; (2) S/S at the same instant of time (VCE) is related to an index from previous work which was defined as a velocity of internal shortening; (3) specific indices of S/S were decreased with lower temperature but increased with time in a Ca-free solution. Our results for a Ca-free solution do not agree with previous work but can be explained by changes of the smooth muscle cell membrane. This suggests that L-independent indices are useful for characterizing changes in the overall dynamic state of the tissue but are not necessarily representative of a change in crossbridge dynamics. PMID- 7470639 TI - The in vivo diffusion chamber technique for bone marrow or blood cell culture. PMID- 7470640 TI - The value of prognostic indices in aplastic anaemia. AB - In 43 patients with aplastic anaemia we assessed the accuracy of different prognostic systems. Patients dying within 6 months after diagnosis were correctly predicted in 60% of cases with the Lynch-index with a sensitivity of 82%. With the Najean-index 40% of these patients are correctly predicted, this index has a sensitivity of 100%. More accurate are the prognostic criteria proposed by Camitta et al [5]. With these criteria, this rapidly fatal group is correctly predicted in 85% of the patients, indicating that 15% of the patients are incorrectly predicted to have a limited survival. The sensitivity, however, is 100%. The Lohrmann-index, based on reticulocyte count predicts 64% of this group with severe aplasia. None of these prognostic systems do accurately predict long survival. We suggest that the best differentiation between patients with a long term prognosis (more than 5 years) and patients who die from aplastic anaemia within 5 years, is made by re-evaluating the leucocyte and platelet count 3 months after the initial diagnosis. Decrease in blood counts (over 10%) predicts death from aplastic anaemia within 5 years correctly in all patients; stable or increased blood counts predict long survival in 75% of the patients. PMID- 7470641 TI - Clinical evaluation of simultaneously administered 58Co labelled vitamin B12 and 57Co labelled vitamin B12 bound to intrinsic factor in patients with pernicious anaemia. AB - The clinical and laboratory data from 75 patients with altered vitamin B 12 absorption were reviewed. In 36 cases the diagnosis of pernicious anaemia had been established. Of these, 14 patients showed malabsorption of radiolabelled vitamin B 12, but the absorption of vitamin B 12 bound to the intrinsic factor (IF) was normal (Group A). The other 22 patients with pernicious anaemia showed altered free and IF-bound vitamin B 12 absorption (Group B). Laboratory and clinical data and the absorption tests (xylose and fat excretion) were more abnormal in group B than in group A. Finally, 39 patients (Group C) with general malabsorption showed alternation of the absorption of free and IF-bound vitamin B 12 (Group C). The clinical nd biological data were different in these patients from that found in groups A and B. It is possible that in patients with pernicious anaemia who are untreated an alteration of the intestinal mucosa may produce a malabsorption of vitamin B 12 even in the presence of intrinsic factor. PMID- 7470642 TI - The efficacy of neuraminidase treatment in studies on red cell aging. PMID- 7470643 TI - Effects of chlorite exposure on conception rate and litters of A/J strain mice. PMID- 7470644 TI - In vitro metabolism of carbofuran by liver microsomes of the padifield fish Trichogaster pectoralis. PMID- 7470645 TI - Effects of cadmium, lead and copper on rat preimplantation embryos. PMID- 7470647 TI - Accumulation of copper in fluvial sediments and yellow water lilies (Nuphar lutea) at varying distances from a metal processing plant. PMID- 7470646 TI - Effects of chlorpropham, chlortoluron and isoproturon on respiration and transformation of nitrogen in two soils. PMID- 7470648 TI - Cardiac and non-cardiac malformations produced by mercury in hamsters. PMID- 7470649 TI - Confirmation of residue identity: characterisation of products of the reaction between chromium (II) solutions and aldrin. PMID- 7470650 TI - A rapid and simple method for the determination of volatile N-nitrosamines in biological materials. PMID- 7470651 TI - Characterization of four commercial flame retardant aryl phosphates. PMID- 7470652 TI - Adaptation of a soil bacterium to hydrazine propellants. PMID- 7470653 TI - Exposure of seed pelleting plant workers to methiocarb. PMID- 7470654 TI - Effect of boron in drinking water on the male laboratory rat. PMID- 7470655 TI - Toxicity of nickel, copper, zinc and aluminum mixtures to the white sucker (Catostomus commersoni). PMID- 7470656 TI - Use of a cell wall-less mutant strain to assess the role of the cell wall in cadmium and mercury tolerance by Chlamydomonas reinhardtii. PMID- 7470657 TI - Comparative study of the acute toxicity of a homologous series of trialkyltins to larval shore crabs, Hemigrapsus nudus, and lobster, Homarus americanus. PMID- 7470658 TI - Simplified monitoring procedures for benzo(a)pyrene, hexachlorobenzene and pentachlorophenol in water. PMID- 7470659 TI - Low erythrocyte glucose-6-phosphate dehydrogenase (g-6-PD) activity and susceptibility to nitrite-induced methemoglobin formation. PMID- 7470660 TI - Toxicity of five forest insecticides to cutthroat trout and two species of aquatic invertebrates. PMID- 7470661 TI - Lead concentrations in livers of Maryland waterfowl with and without ingested lead shot present in gizzards. PMID- 7470662 TI - An ultraviolet detector for automated size exclusion chromatographic cleanup of lipid-containing residue samples. PMID- 7470663 TI - Heterogeneous hydrodechlorination of toxaphene. PMID- 7470664 TI - Mercury accumulation in biota of Thunder Creek, Saskatchewan. PMID- 7470665 TI - Bioaccumulation of hexachlorobenzene in killifish (Fundulus similis). PMID- 7470666 TI - Dissimilarities in the toxic response of early chick embryos to DDT administered in different vehicles. PMID- 7470667 TI - Distribution of radioactivity in the chondrichthyes Squalus acanthias and the osteichthyes Salmo gairdneri following intragastric administration of (9 14C)phenanthrene. PMID- 7470668 TI - Toxicity of nitrite to larvae of the salamander Ambystoma texanum. PMID- 7470669 TI - Toxicities of paraquat and diquat herbicides to freshwater copepods (Diaptomus sp. and Eucyclops sp.). PMID- 7470670 TI - Toxicities of eight organochlorine compounds in sediment and seawater to Crangon septemspinosa. PMID- 7470671 TI - Avoidance response of juvenile Chromis punctipinnis to chlorinated seawater. PMID- 7470672 TI - Heavy metal concentrations in feathers of ruffed grouse shot by Virginia hunters. PMID- 7470673 TI - Lethality of permethrin, cypermethrin and fenvalerate to salmon, lobster and shrimp. PMID- 7470674 TI - [Sensitivity to tuberculin in Swedish schoolboys vaccinated with BCG at birth]. PMID- 7470675 TI - [Early diagnosis of chronic bronchitis, study with the point of view of public health]. PMID- 7470676 TI - [Possible causes of the failure of the treatment of pulmonary tuberculosis ; how to avoid them]. PMID- 7470677 TI - Medical library service in a community-based medical school: a case study in South Dakota. AB - The historical background of community-based medical schools is described with emphasis on the experiences of the University of South Dakota Lommen Health Sciences Library. The steps undertaken by the library to meet Liaison Committee for Medical Education accreditation standards required for a full four-year, M.D. degree granting institution are outlined. The governance structure of the participating Libraries of the Affiliated Teaching Hospitals Council is described. Special problems and their solutions are discussed in the context of providing service to a medical school which is decentralized on a statewide basis. PMID- 7470678 TI - Continuing education in cancer for the community physician: design and evaluation of a regional table of contents service. AB - The Arnold Library of the Fred Hutchinson Cancer Research Center offered two groups of practicing physicians in nonmetropolitan areas of the Pacific Northwest an opportunity to review regularly their choice of eighteen cancer journal tables of contents with the added provision (in compliance with federal copyright laws) of being able to receive any articles of interest at no charge from the library. Forty-nine (38.9%) of the 126 physicians selected chose to participate in the program. Of these, 20 (40.8%) actually requested articles from the service. After six months of the service, all of the original 126 physicians were queried by questionnaire. Those who used the service felt it helped in their practice of medicine and that it should be continued. Those who chose not to receive the service generally had access to the eighteen journals elsewhere or saw very few cancer patients. A majority of the articles which had been requested emphasized patient treatment. Approximately 25% of one library technician's time and $360 for photocopy and mailing costs were required to operate the service for forty nine physicians for six months. The service seemed to identify a select audience of regular users who perceived a benefit from its use. PMID- 7470679 TI - The application of selected evaluative measures to the library's monographic ophthalmology collection. PMID- 7470680 TI - An examination of characteristics related to success of friends groups in medical school rare book libraries. AB - Friends of the library groups traditionally have been considered effective sources of funding for libraries. Empirical evidence was sought to determine the accuracy of this belief for medical rare book libraries. Characteristics of the medical rare book library and of the friends group were examined to identify those which are correlated with successful friends groups. Correlations were found between the amount of money contributed by the friends group and the age of the friends group, the librarian's participation in forming the group, and the amount of money spent by the library on the friends group. Recommendations are made toward a goal of larger donations from friends groups of medical rare book libraries and toward more effective management of those funds. PMID- 7470681 TI - [Invasive meningiomas of the sphenoid ridge]. PMID- 7470682 TI - Evaluation of a new piston-type spirometer. AB - A new dry spirometer (DS) incorporating a microcomputer for automatic calculation and direct read-out of spirometric parameters was compared to a calibrated Fleisch pneumotachograph (PT). Twenty-eight volunteers, including healthy subjects, obstructive and non-obstructive patients, performed a total of 48 spirometric tests with the two apparatuses randomly alternated. DS values for FVC were lower than those of the PT with a mean difference of 0.177 1 +/- 0.171 SD (p less than 0.0001). Mean FEV1 values were very similar in both apparatuses with a non significant difference of 0.003 1 +/- 0.182 FEF25-75 values were higher with the DS, showing a mean difference with the PT of 0.407 1 . s-1 +/- 0.324 (p less than 0.00001). The DS also gave slightly higher PEFR values than the PT, with a mean difference of 0.310 1 . s-1 +/- 0.830 (p less than 0.02). Differences in FVC readings between DS and PT were greater in severely obstructed patients (0.350 1 +/- 0.084, p less than 0.0001) than among healthy subjects (0.120 1 +/- 0.051, p less than 0.0002). On the contrary, in absolute figures DS values for FEF25-75 showed a greater difference with those of the PT in normals (0.547 1 . s-1 +/- 0.320, p less than 0.001) than in obstructed patients (0.288 1 . s-1 +/- 0.244, p less than 0.05). DS flow resistance was 2.67 cmH2). s. 1-1 for a flow of 8.8 1. s 1. Based on the spirometric differences observed, and on the high flow resistance and volume limitation imposed by its 6 1 capacity, we concluded that the apparatus tested cannot be recommended for clinical spirometry. PMID- 7470683 TI - Respiratory drive and ventilatory pattern during exercise in interstitial lung disease. AB - Ventilation was analysed during a maximal and progressive exercise in 12 patients suffering from interstitial lung disease (ILD) and in 15 healthy subjects. "Rapid shallowing breathing" pattern was observed in patients with ILD. Both normals and patients showed a linear relationship between VE, VT/TI (mean inspiratory flow) and VCO2. The relationship between P0.1, the occlusion pressure at 0.1 S, and VCO2 exponential in both groups and significantly steeper in patients with ILD. P0.1/VE and P0.1/(VT/TI) ratios, indexes of thoracopulmonary impedance, and P0.1/VCO2 ratio, an index of respiratory drive per unit of CO2 output, were all inversely correlated in patients with VC% (vital capacity) and CLstat (static compliance). These results strongly suggest that in patients suffering from ILD the higher respiratory drive observed during exercise could be attributed to increased afferent reflexes originating from the lung and/or chest wall. PMID- 7470684 TI - The effect of diffuse pulmonary fibrosis on lung mechanics. AB - Maximum expiratory flow-volume curves (MEFV), inflational and deflational pressure-volume curves, dynamic compliance at 20, 40, and 60 breaths per minute, closing volume (single-breath oxygen method), standard lung volumes are airway resistance measurements were made in eleven subjects with a clinical diagnosis of diffuse interstitial fibrosis (DIF) and eleven control subjects. No subject was a current cigarette smoker. Maximum flow-static recoil pressure curves (MFSR) were constructed and calculations of upstream resistance (Rus) were made. Comparison of the mean values showed no significant differences in airway resistance or FEV1/FVC% between the two groups; however, significant differences were noted in total lung capacity (TLC), forced expiratory volume in one second (FEV1) and expiratory flow rates. Nine of the diseased subjects had pressure-volume curves shifted to the right of the normal range, and a significant decrease in static expired and inspired compliance was noted in the diseased group. Six subjects with DIF showed increased Rus. The single-breath oxygen test results revealed an increased slope of phase III (delta N2/I) in the diseased group. Decreases in the static compliance may be explained by loss of lung volume in some patients with DIF. There is also evidence that the small airways may be affected by this disease process. PMID- 7470685 TI - COHb formation and acute carbon monoxide intoxication in adult male rats and guinea-pigs. AB - Respiratory exchange recordings and percentage survivals to lethal concentrations show that rats are more sensitive than guinea-pigs to an acute carbon monoxide intoxication. To avoid circadian respiratory an comportmental differences between rats and guinea-pigs experiments were performed on urethane anesthetized and artificially ventilated animals. This procedure demonstrates that, for an inhaled concentration of 0.05% CO, the rates of formation of COHb do not statistically differ in rats and in guinea-pigs, but that a higher (0.01 less than p less than 0.05) COHb saturation is reached in rats (35%) than in guinea-pigs (25%). For a 2.84% CO inhalation, no statistically significant difference is observed in the rate of COHb formation, but cardiac arrest is sooner (p less than 0.001) observed in rats than in guinea-pigs. PMID- 7470686 TI - [Influence of sequential breath-holding on the slope of phase III (author's transl)]. AB - Many factors contribute to the slope of phase III of the single-breath nitrogen test (SB-N2). By its effect of nitrogen concentrations, alveolar oxygen exchange may be responsible for a fraction of this slope. In fact, the slope of phase III is steeper when expiratory time is prolonged. We studied the effect of two sequential breath-holding periods of 20 seconds each with ten healthy young male volunteers, by SB-N2 and a reverse technique (SB-R). We observed a sudden increase (mean 1.27%) in the expired nitrogen concentration after each breath holding period. Closing volume remained the same in all tests, but the slope of phase IV decreased by 44% in SB-N2A (SB-N2 with breath-holding) and increased by 55% in SB-RA. Expiratory vital capacity was significantly less after breath holding. These data suggest that gas exchange during expiration is partly responsible for the slope of phase III. PMID- 7470687 TI - Emphysema in smoking and non-smoking coalworkers with pneumoconiosis. AB - The relationships between coalworkers' pneumoconiosis, cigarette smoking and emphysema in a group of 139 deceased coalminers, 19 of whom were non or ex smokers of 10 years standing, were studied from available clinical and morphological evidence. There was 95 cases of simple pneumoconoiosis and 44 cases with early (Cat. A) progressive massive fibrosis (PMF). The objects were to compare the amounts and types of emphysema in the smokers and non-smokers and also their ventilatory findings. The emphysema content of all the 139 cases of smokers and non-smokers was measured by a standard method and the amounts compared. In addition, the emphysema content of the 19 non- and ex-smokers plus 32 appropriate control cases drawn from the smoking group were measured afresh by a new method devised by the pathologists for the estimation of the extent and type of both emphysema and coalworkers, pneumoconiosis, from large paper lung sections and histological preparations. Both methods compared well in estimating emphysema content and both indicated that the amounts of emphysema in the smoking and non-smoking groups were similar. These findings still applied when both the life-long non-smokers and the cases with simple pneumoconiosis only were looked at separately. It was found that centrilobular emphysema was much commonest type encountered in both smokers and non-smokers. The non-smokers were less disabled in terms of impairment of FEV1.0 than the smokers, although the difference was not statistically significant. The relationship between the centrilobular emphysema and coalworkers' pneumoconiosis is discussed. PMID- 7470688 TI - Retention of particles on the first bifurcation and the trachea of rabbits. AB - Retention of 6 micron, intratracheally-administered radioactively-tagged, teflon particles on the lower trachea and the first bifurcation of rabbits was measured externally for 2-3 days. Clearance occurred in a fast and a slow phase. The fast phase was finished after 1-2 h. The slow phase differed markedly between the lower trachea and the first bifurcation. The mean half-times were 19 and 39 h i trachea and first bifurcation, respectively. On average, 39% of the particles on the trachea and 71% on the first bifurcation cleared with the slow phase. Thus the bifurcation is more exposed to pollutants not only from deposition of more particles caused by impaction but also because of less efficient clearance. PMID- 7470689 TI - [Mechanical impedance of the chest in smokers and non-smokers (author's transl)]. AB - Mechanical impedance of the respiratory system was measured at 5, 10, 15 and 20 Hz in 47 healthy non-smokers, 20 ex-smokers and 51 current smokers. Besides the resistance and its frequency dependence, the compliance, the inertance and the resonant frequency of the respiratory system were computed using the assumption of a second order linear system. The data were normalized for sex, age and body height on the basis of correlations observed in non-smokers. No significant difference was found in ex-smokers. In smokers the resistance was significantly increased at all frequencies. However, it was normal in a number of subjects with abnormally low maximal expiratory flows. unlike maximal flows, forced oscillations indices were not correlated to tobacco consumption expressed in pack years. The results do not indicate that impedance measurements breathing air may be specially useful for early detection of airway abnormalities. PMID- 7470690 TI - [Freedom of the patient in medicine today]. PMID- 7470691 TI - [The dignity and rights of the patient]. AB - In his treatment of the patient, the doctor must be guided not simply by concern for the patient's biological existence but ultimately by concern for the dignity and integrity of his person. These qualities are founded in the patient's innate human freedom, which equally underlies his entitlement to life and to proper treatment without sacrifice of his right to self-determination, his inviolability and his human dignity. Self-determination presupposes the right to information and communication, and this in turn an obligation on the part of the doctor to talk in terms which the patient understands and to devote sufficient time to his needs. Respect for the inviolability of the patient's person implies not only his right to the security of confidentiality and discretion, but also his right, when receiving any form of treatment, to be acknowledged as a physiological, psychological and social entity with a concrete individuality. Human dignity, finally, implies the right of each individual to be the real "raison d'etre" for any treatment to which he is subjected. Beyond this, a right to health can only exist in the negative sense of the right not to be made avoidably ill by the social order, and a right to sickness only in the restricted sense of not being put at a social disadvantage as the result of a medically diagnosed illness. PMID- 7470692 TI - [The dignity of the patient]. PMID- 7470693 TI - [The dignity of the patient and medical progress from the Catholic point of view]. PMID- 7470694 TI - [The dignity of the patient: a legal problem?]. AB - Rules of medical ethic bid since the times of antiquity to respect the patients dignity. Today to respect the dignity of any man is a general legal rule. In the same way other rules of the traditional medical ethic became legal norms as for instance the command forbidding to do harm to someone. The law intends to limit the possibilities to exercise power. Therefore it does not compete to the doctor alone to decide that a medical treatment has to be applied. The legal base of a medical treatment lies in the consent of the patient who has been duly cleared up on his state, the necessary treatment and its risks. If it has to be decided if a doctor has given the right treatment in a special case it does not suffice to consult the general rules of the law; the circumstances of the case have to be considered as well. The doctor has to decide, according to the actual medical knowledge and the rules of his professional art what the appropriate proceeding is. Legal and medical considerations are closely connected if one judges a doctor handling a special case. If the patient consents, the doctor is not obliged to treat him, but he is entitled to do it, cases of emergency excepted. If and in what way he treats the patient has to be decided by the doctor according to medical criterias. If a patient, sound of mind, who is suffering heavily by an incurable illness asks the doctor to restrain treatment to alleviating the pains and to the absolute cares to preserve life, the doctor is bound by his patient's wish. In analogy the legal construct of "conducting business without mandate" allows the doctor to proceed in the same way if the patient who lost consciousness is not able to decide upon the treatment and whose death is inevitable and imminent if this is the only wise to respect the dignity of the patient. PMID- 7470695 TI - [Respect for the patient in the face of medical progress]. AB - The psychological aspects of medical ethics are described and analysed in terms of the doctor-patient relationship. Analysing the patient's needs, the doctor ceases to regard him as an "object" for his diagnostic and therapeutic procedures and makes him the "subject"; this implies respect for his person and his rights. From this psychological and ethical perspective, questions of patient information, professional secrecy, therapy transfer and the doctor's authority are discussed. Psychiatry poses specific problems both in practice (admission of patients to hospital although their illness prevents them from giving their consent) and in its theory (is mental illness real?). Ethical problems force the practitioner to reconsider his concept of man and his options in respect of freedom. PMID- 7470696 TI - [Introduction to the "Guidelines of Medical Ethics" of the Swiss Academy of Medical Sciences]. AB - The guidelines of medical ethic are recommendations of the Swiss Academy of Medical Sciences to the medical doctors to overcome the difficulties of medicine and modern technics and social problems. In an introduction the duties of the physician and his behaviour in the hippocratic mind are exposed, without asking for an oath. The recommendations give I. essential rules of international health organisations and II. Medical-ethic guidelines regarding different questions: 1. research examinations on men 2. euthanasia 3. transplantation 4. artificial insemination 5. sterilisation, especially the operative sterilisation of mentally handicapped persons, without acceptation of the so-called forced sterilisation 6. medical-ethical committees. The newly founded medical-ethical commission of the Swiss Academy of Medical Sciences and its organisation with physicians and laymen is explained. Its different functions are: a) answers to inquiries of international or national organisations and of individual persons b) formulation of principal ideas for medical-ethical questions, which will arise also in the future, not only on research, but also on diagnostic, therapeutic or prophylactic problems. These medical-ethical guidelines are formulated in few words to help the medical doctor for a quick consultation. They give a legal ethic. The Swiss Academy of Medical Sciences considers this as one of her tasks: to guard and to guard and to advance ethical devices in medicine. PMID- 7470697 TI - [Ethics and the Federation of Swiss Physicians]. AB - As a professional organization, the Federation of Swiss Physicians must make sure that generally admitted rules of ethics as well as professional regulations in this matter are recognized and observed by its members. In its statutes, it asserts itself for a high level medical care of the population; it requires not only a permanent renewal of professional aptitude as a duty in the widest meaning of the word, but also a great sense of responsibility for patients and society. Axioms on professional ethics are unfit as arguments for material requirements to fulfil the best possible all these duties. Credibility is only granted if ethical rules are really observed. PMID- 7470698 TI - [Ethics and the physician's practice]. AB - Although a doctor may not be continually aware of it, his medical activity is firmly rooted in the moral principles of the medical profession. Serious problems of medical ethics may arise anywhere at any time from a general practitioner's Cases - and may lead to difficult decisions. Consequently medical men have from the early days felt the need to commit their ways of thinking and acting to a code of laws, which they can consult whenever taking a difficult decision. The doctors' persistent efforts to find an unambiguous doctrine have been evident from the Hippocratic oath up to the "Guidelines concerning Assistance to the Dying" of the Swiss Academy of medical Sciences. PMID- 7470699 TI - [Medical ethics problems in psychiatry]. AB - A humanitarian attitude and ethical convictions have contributed much to the development of psychiatry. In the present time these convictions have been codified, for instance in the declaration of the World Federation of Psychiatry in Hawaii 1977. In more detail the author discusses 3 areas where ethical considerations have special significance: the scope of psychotherapy, psychosurgery, and the use of psychiatric means to suppress and discipline politically dissident persons. PMID- 7470700 TI - [Ethics problems in neonatal medicine]. AB - In view of the recent advances in reanimation of the newborn infant, parents, physicians and nursing staff have now to face the ethical dilemma of the quality of the surviving infant with severe brain damage which excludes a further normal development. The author discusses the situation of an infant with trisomy 21 and a total intestinal atresia: a surgical intervention leads to a living human being with a severe mental, and physical handicap; a non-intervention will let the newborn die even if he could be saved. The decision has to be made with the parents in agreement with the nursing staff, but without involving a policy making outside committee. PMID- 7470701 TI - [Attempt to define an ethical code for prenatal diagnosis]. PMID- 7470702 TI - [Medical ethics in prison medicine]. AB - A series of situations and decisions involving medical ethics in a prison medical service are discussed. The doctor's independence is considered in relation to his contract with administrative authorities. In contrast with most private doctor patient relationships, there is usually no possibility for prisoners to choose their doctor and vice-versa. Freedom of consent on the part of the patient may also imply a right to no-treatment. Medical care in prison is not easy to delineate, also because patients often try to involve the doctor in non-medical demands. A prison doctor should avoid taking part in decisions which ought to be made by the judiciary or by administrative authorities. Programmes involving preventive medicine and sociotherapy imply collaboration between therapeutic and security staff. The continuous interplay and readjustment between powers based on public authority, on the rights of each individual prisoner and on the medical programmes makes it possible for some sort of therapeutic freedom to exist in the prison. PMID- 7470703 TI - [Medical ethics and terminal care]. AB - Numerous researches have been done on this subject. The Swiss Academy of Medical Sciences as well as the European Council have prepared guidelines that are very clear. However what is lacking is an adequate preparation of the physicians and nursing personnel concerning attitudes. This inadequate preparation is a major reason for the violation of the dying patients rights. Accompanying the dying and his family requires on extensive knowledge of oneself and especially of the patient. It also requires patience, honesty, trust, humility, hope and courage. The mentioned characteristics, described by the American philosopher Milton Mayeroff, are indispensable to the professional who really wants to "care" for the other person. Today, we need to provide specific training in order for young physicians to develop these characteristics. PMID- 7470704 TI - [Medical ethics and the public health office]. AB - The Federal Office of Public Health is a governmental administration in charge of control and coordination measures related to the field of health protection. A rough 20% of the 220 officials and employees have university education prevailingly in medicine or natural sciences. Any government is working for the benefit of the general public and ought not to end in itself. Although there is a potential danger of abuse of official authority and power and of corruption, such deviations have proved to be quite rare in Switzerland. The safety guarantees are sufficient and we furthermore dispose in general of a professionally and morally well qualified staff. A governmental institution is as good or as bad as the humans working for it. A careful selection of the civil servants is therefore of utmost importance. On the whole, the quality of our civil service is quite superior to its reputation. Negative global judgements are to be declined. The Federal Office of Public Health aims at respecting the principles of the medical ethics in all its activities. PMID- 7470705 TI - [The role of mucopolysaccharides in the regulation of ocular pressure: morphometric study]. PMID- 7470707 TI - [Experimental herpetic keratitis, in vivo and in vitro study with the scanning microscope]. PMID- 7470706 TI - [Melanocytoma of the optic papilla]. PMID- 7470708 TI - Glucosamine therapy: a preliminary note on a new approach in treatment of herpetic keratitis. PMID- 7470709 TI - [Complications in 1000 consecutive intracapsular cataract extractions]. PMID- 7470710 TI - Pseudo-exfoliation of the lens capsule (exfoliation syndrome) after iridocapsular lensimplantation. PMID- 7470711 TI - [Ocular hypotension and cataract in pregnancy, apropos of a case]. PMID- 7470712 TI - [Absorbable sutures and aphakic astigmatism]. PMID- 7470713 TI - [Fibro-xanthomatous invasion of the cornea treated with intrastromal keratectomy]. PMID- 7470714 TI - Chemotherapy for gastrointestinal cancer. PMID- 7470715 TI - The combined approach. PMID- 7470717 TI - Violent behaviour by psychiatric patients. AB - There is no substantial body of evidence to prove that psychiatric patients as a group are more violent than the general population. In determining whether violence is exhibited, the underlying personality of the individual is far more important than the psychiatric illness from which he suffers. Individuals with psychopathic personalities, those who abuse or are dependent on alcohol drugs, and those with pronounced paranoid or sadistic traits may be characterized by aggressive and violent behaviour. There is at present no reliable method of predicting if or when an individual patient will exhibit assaultive behaviour. Psychiatric hospitals, especially in the UK, are characterized more by frequent petty violence than by serious violence. Victims of assaults by patients are often related or known to them an interact with them on a personal level; they may also play a considerable role consciously or otherwise in instigating the assault. PMID- 7470716 TI - Antibiotics for gram-positive organisms. AB - Most infections due to Gram-positive organisms can be treated with quite a small number of antibiotics. Penicillin, cloxacillin, and erythromycin should be enough to cover 90 per cent of Gram-positive infections. The relatively narrow spectrum of these drugs should be the incentive to prescribers to use them selectively, together with adequate bacteriological investigation, in order to achieve effective treatment with a minimum of disturbance to the patient's normal bacterial flora and without any other harmful side effects. PMID- 7470719 TI - Disease of the month. PMID- 7470718 TI - Depression in the elderly. AB - The elderly are particularly susceptible to serious depressive illness. While physical disease and social deprivation might seem to explain this susceptibility there is still much to be learned about the aetiology, especially with regard to organic cerebral factors. Response to treatment in the short term is good, but the long-term outcome is one of high morbidity demanding an extensive commitment from psychiatric and social services. PMID- 7470720 TI - The French connection. PMID- 7470721 TI - Recent developments in infusion devices. PMID- 7470722 TI - Blood sugar levels. PMID- 7470723 TI - Exercise testing in ischaemic heart disease. AB - The exercise test may be regarded as an objective measure of functional capacity and a useful adjunct to the history given by the patient. As a means of diagnosis it has many limitations but a positive result in a male patient provides good corroborative evidence for the presence of coronary artery disease. As a means of predicting severity of disease the test remains imperfect but may prove useful with further refinements. The exercise test is of little value for screening except in special circumstances. PMID- 7470724 TI - How to manage a cardiac arrest. PMID- 7470725 TI - Buying your own home. PMID- 7470726 TI - Correlation and linear regression. PMID- 7470727 TI - Leucine-enkephalin and methionine-enkephalin produce opposing effects on plasma corticosterone levels in ether-stressed mice. AB - The inhibitory effects of intracerebroventricular administration of saline on the plasma corticosterone response to ether stress in mice was reduced by Met enkephalin and enhanced by Leu-enkephalin. When administered simultaneously the effects of the two peptides opposed each other. Met and Leu-enkephalin may subserve different physiological functions in the response of the hypothalamus pituitary-adrenal system to stress. PMID- 7470728 TI - Exocytotic release of catecholamine from perfused adrenal gland of guinea-pig induced by veratridine. AB - 1 Experiments were carried out on perfused adrenal glands of guinea-pig to determine whether veratridine caused the exocytotic release of catecholamine by comparing its effect with that of splanchnic nerve stimulation and secretagogues such as acetylcholine and excess K+. 2 Veratridine (100 microM) and excess K+ (56 mM) caused secretion of catecholamine and dopamine-beta-hydroxylase (DBH) activity in the venous effluents in the presence of atropine (30 microM) and hexamethonium (2 mM). Splanchnic nerve stimulation in the presence or absence of physostigmine (100 nM) and infusion of acetylcholine in the presence of physostigmine had the same effect. In all the responses, the release of DBH tended to last for a longer period than that of catecholamine. 3 The ratio of catecholamine to DBH activity appearing in the venous effluents was approximately 9, regardless of the method of stimulation. This value was close to the ratio of catecholamines to the 'soluble' DBH activity found in the chromaffin granules. 4 All the types of stimulation used caused a proportional release of adenine nucleotides and catecholamines in the effluents. The adenine nucleotides were mainly adenosine 5'-phosphate. 5 The ratio of catecholamine to adenine nucleotides was approximately 11, regardless of the method of stimulation. 6 It is suggested that the release of catecholamine induced by veratridine occurs by exocytosis in adrenal glands of guinea-pig. PMID- 7470730 TI - Further studies on the inhibition of monoamine synthesis by monofluoromethyldopa. AB - 1 alpha-Monofluoromethyldopa (MFMD, RMI 71963), a potent and selective enzyme activated irreversible inhibitor of aromatic L-amino acid decarboxylase produces a substantial and long-lasting decrease in the catecholamine content of mouse brain, heart and kidney. 2 Single doses of MFMD reduce the 5-hydroxytryptamine concentration of mouse brain without altering the tryptophan concentration. 3 In animals treated with MFMD, peripheral but not brain noradrenaline is restored within 1 h to control levels by an intraperitoneal injection of dopamine. PMID- 7470729 TI - Effects of adrenalectomy and hypophysectomy on enkephalin content of the rat hypothalamus. AB - 1 Hypothalamic content of enkephalin in rats has been measured by bioassay against methionine-enkephalin on field-stimulated mouse vas deferens after acid extraction and purification using Amberlite XAD-2 resin.2 Surgical stress lowered hypothalamic enkephalin content initially but 6 days after operation the content was higher in sham-operated than in adrenalectomized animals.3 Corticosteroid replacement therapy showed that hypothalamic enkephalin content was not related directly to circulating corticosteroid levels and that it was increased by the stress of handling and injection.4 Hypothalamic enkephalin content of rats that had undergone hypophysectomy 11 days earlier did not differ from that of intact animals but surgical stress, which lowered content in intact rats and had no effect after adrenalectomy, produced a rise in content in these hypophysectomized animals.5 The ingestion of 0.9% saline, in place of water, by sham adrenalectomized rats altered the effect of surgical stress on hypothalamic enkephalin content without affecting the resting levels.6 Of extracts from adrenalectomized rats, 10% contained a substance that behaved atypically on the mouse vas deferens, showing a slower onset and offset of action. The mol. wt. of the substance and sensitivity to naloxone reversal appeared to be similar to enkephalin, but it was resistant to carboxypeptidase-A and protease treatment.7 It is concluded that the enkephalin content of the hypothalamus is affected by activity in the hypothalamus-pituitary-adrenal system but that it is not related in a simple manner to the levels of corticotrophin releasing hormone, corticotrophin or corticosteroids. PMID- 7470731 TI - Intracellular observations on the effects of muscarinic agonists on rat sympathetic neurones. AB - 1 Responses of single neurones in isolated superior cervical ganglia of the rat to muscarinic agonists were recorded with intracellular microelectrodes. 2 (+/-) Muscarine (1 to 10 microM) and methylfurmethide (1 to 3 microM) produced reversible membrane depolarizations (less than or equal to 15 mV) accompanied by a fall in input conductance and an increased tendency toward repetitive spike discharges. The spike configuration was unchanged. 3 Analysis of steady-state current/voltage curves revealed the most consistent muscarinic effect to be a large reduction (approximately 50% at 10 microM muscarine) in input slope conductance around rest potential. This conductance decrease diminished as the membrane was hyperpolarized, and the normal increase in slope conductance with membrane depolarization was depressed. The current/voltage curves in the between 65 and -88 mV (i.e. 9 to 28 mV hyperpolarized to rest potential). 4 Divalent cations (10 mM [Ca2+] or [Mg2+]) showed a small muscarine-like effect on the current/voltage and slope conductance/voltage curves, but did not affect the action of muscarine itself. 5 Tetraethylammonium (TEA, 5 mM) also had a small muscarine-like effect, and depressed or reversed the action of muscarine. However, TEA differed from muscarine in blocking orthodromic transmission and prolonging direct spike repolarization. 6 It is concluded that the primary effect of muscarinic agonists is to alter the rectifying properties of the cell within the potential range -80 to -40 mV. PMID- 7470732 TI - Response of human isolated bronchial and lung parenchymal strips to SRS-A and other mediators of asthmatic bronchospasm. AB - 1 The responses of human isolated bronchial and lung parenchymal strips to cumulative doses of slow reacting substance of anaphylaxis (SRS-A), histamine, prostaglandin F2 alpha (PGF2 alpha) and acetylcholine have been examined, after storing the tissues overnight in Krebs solution at 4 degrees C. 2 Both tissues contracted to all four agonists. The order of potency (as determined by height of maximal contraction) was: bronchial strip: acetylcholine greater than histamine = PGF2 alpha greater than SRS-A, and parenchymal strip: PGF2 alpha much greater than histamine = SRS-A greater than acetylcholine. 3 Maximal contractions to SRS A of both the human bronchial and parenchymal strips were approx. 30% of the maximal contractions produced by the most potent agonist on each tissue (PGF2 alpha on the parenchymal strip and acetylcholine on the bronchial strip). SRS-A, therefore, does not have a powerful direct contractile effect on either parenchymal or bronchial strip of human lung, and is approximately equipotent on both tissues. A part of the broncho-constrictor activity of SRS-A in vivo may be mediated via indirect pathways. 4 The selective SRS-A antagonist, FPL 55712, was approximately equipotent in antagonizing contractions induced by SRS-A on both human bronchial and parenchymal strips. PMID- 7470734 TI - The size of hydroxyl groups in solution and the changes in size associated with the ionization of phenolic, carboxylic and amino groups in phenolic quaternary ammonium salts, nicotine and some amino acids: possible implications for drug water and drug-receptor interactions. AB - Size in solution can be expressed either as the apparent molal volume at infinite dilution (phi 0v) amd the concentration parameter (j) or as the partial molal volume of the solute at infinite dilution (V0(2)) and the concentration parameter for the solute or solvent (qs or qw). Although calculated differently, these are derived from the same results and are equivalent. From measurement with phenolic quaternary ammonium salts, including compounds with high nicotine-like activity, the apparent size of the hydroxyl group in water is small and variable. Phenolic groups are slightly larger than alcoholic groups, which should be better hydrogen donors. By measuring the volume change associated with ionisation it is possible to measure the size of charged groups such as phenate and carboxylate; these are much smaller than phenolic and carboxyl. Ammonium groups, however, are only slightly smaller than the corresponding amines. The zwitterion forms of amino acids are associated with a minimum in volume but the volume changes increase with chain length from glycine to gamma-aminobutyric acid. Groups separated by less than this distance interact in their effects on water. Decreases in volume or unexpectedly small increments in apparent molal volume represent decreases in entropy which must be taken into account in drug-water-receptor interactions. Although they may be offset by enthalpy changes, they should favour binding because there is more scope for an increase in entropy. This might explain the association of the small apparent size in water of the hydroxyl group in many compounds with its effects of their affinity for receptors. PMID- 7470733 TI - Absence of changes in drug disposition and catecholamine sensitivity in the hyperthyroid dog. AB - 1 In order to study the relative contribution of hepatic drug metabolizing enzymes and hepatic blood flow to the clearance of drugs in the hyperthyroid state, the disposition kinetics of two model compounds (antipyrine and propranolol) were examined in thyroid-fed dogs as compared to euthyroid and phenobarbitone-pretreated animals. 2 In hyperthyroid dogs, the possibility of catecholamine hypersensitivity was evaluated by assessing the chronotropic response to isoprenaline and by constructing a drug concentration-effect (beta blockade) relationship. 3 The plasma propranolol half-life (0.97 +/- 0.12 h) of the hyperthyroid animals did not differ significantly from either the euthyroid group or the phenobarbitone-pretreated group. This was observed with no significant change in the apparent volume of distribution among the three experimental groups. 4 Phenobarbitone pretreatment accelerated significantly the elimination of antipyrine (half-life, 1.09 +/- 0.15 h, P less than 0.01) as compared to the euthyroid (2.84 +/- 0.35 h) and the hyperthyroid groups (2.58 +/- 0.13 h), respectively, without any changes in the apparent volume of distribution in any group. 5 Neither the chronotropic responses to exogenously administered catecholamine, nor the antagonist concentration-effect relationships support the concept that the hyperthyroid state potentiates sensitivity of the receptor effect system of the heart. 6 The data obtained from the present study fit best with the view that thyroid hormone excess alters neither the disposition of the model compounds used nor the catecholamine-sensitivity examined. PMID- 7470735 TI - Mechanism of action of dopamine on the guinea-pig gastro-oesophageal junction in vitro. AB - 1 The effect of dopamine on longitudinal muscle strips of the guinea-pig isolated gastro-oesophageal junction was compared with the response obtained to phenylephrine, isoprenaline and clonidine. Phenylephrine (5 x 10(-7) to 5 x 10( 5) M) produced a dose-related contraction, whilst dopamine (10(-6) to 10(-4) M) and isoprenaline (5 x 10(-7) to 2 x 10(-5) M) produced dose-related relaxations. Clonidine was ineffective in doses up to 10(-5) M. 5-Hydroxytryptamine (5-HT) produced a contraction. 2 Phenylephrine was antagonized by alpha 1-adrenoceptor antagonists but unaffected by beta-adrenoceptor antagonists, whilst the opposite was the case for isoprenaline. A mixture of alpha- and beta-adrenoceptor antagonists was required to inhibit completely dopamine-induced relaxations. 5-HT (3 x 10(-7) M) was specifically antagonized by methysergide (3 x 10(-6) M). 3 pA2 values for a range of alpha-adrenoceptor and dopamine receptor antagonists were determined against dopamine and phenylephrine. The relative order of potency of the antagonists was the same for both antagonists and was prazosin greater than spiroperidol greater than phentolamine greater than domperidone greater than haloperidol, with pimozide and metoclopramide being inactive. 4 Tyramine caused dose-related relaxations of the gastro-oesophageal strips which were susceptible to the same range of antagonists as dopamine. 5 Cocaine (6 x 10(-6) M) and desmethylimipramine (3 x 10(-7) M) reduced the relaxations induced by dopamine and tyramine but there were quantitative differences in the antagonism. 6 Tissue from reserpine pretreated guinea-pigs was insensitive to tyramine but the response to dopamine was only partly reduced. 7 Histological examination of the strips revealed the presence of smooth muscle but only a sparse adrenergic innervation. 8 The results suggest that dopamine acts partly indirectly and partly directly on postjunctional alpha- and beta-adrenoceptors. There is no evidence for an action on specific dopamine receptors. PMID- 7470736 TI - Action of enkephalin analogues and morphine on brain acetylcholine release: differential reversal by naloxone and an opiate pentapeptide. AB - 1 Methionine (Met)-enkephalin, leucine (Leu)-enkephalin and their synthetic analogues were tested for effects on the spontaneous release of cortical acetylcholine (ACh) in vivo. The ability of naloxone to reverse the action of enkephalins on ACh release was compared with its action against morphine. An enkephalin analogue, structurally related to Met-enkephalin, was tested for opiate antagonistic activity in ACh release experiments. 2 Intraventricular administration of Met-enkephalin, Leu-enkephalin, D-Ala2-Met5-enkephalinamide (DALA) and D-Ala2-D-Leu5-enkephalin (DALEU) produced a dose-related inhibition of cortical ACh release. Met- and Leu-enkephalin were very similar both in their potency and the time course of their action on ACh release. Both DALA and DALEU were more potent and had a longer duration of action than Leu-enkephalin. Systemic injections of two pentapeptides, D-Met2-Pro5-enkephalinamide and D-Ala2 MePhe4-Met5(O)-ol-enkephalin (33,824), produced a sustained inhibition of cortical ACh release. 3 Naloxone, administered systemically following the depression of ACh release induced by either intraventricular injections of enkephalins (DALA or DALEU), or systemic injections of enkephalins (D-Met2-Pro5 enkephalinamide or 33,824), reversed this depression and restored the release to baseline levels. The effect of D-Met2-Pro5-enkephalinamide on the release of ACh was reversed by naloxone with difficulty. Naloxone also reversed the inhibitory effect of systemic morphine and this reversal was associated with a large overshoot of ACh release. The latter was never observed in the enkephalin experiments. 4 Intraventricular injection of the pentapeptide, D-Ala2-D-Ala3-Met5 enkephalinamide (TAAPM), at doses that did not influence the basal ACh release, blocked or reversed the inhibitory effect of morphine on this release. This peptide did not block the effect of the non-opiate, chlorpromazine, under similar conditions. In two experiments TAAPM failed to reverse the inhibition of ACh release produced by systemically injected enkephalin, D-Met2-Pro5 enkephalinamide. 5 Effects of morphine and enkephalin on ACh release are discussed in terms of their action on difference opiate receptors. PMID- 7470737 TI - Investigation of action of enkephalin on the spontaneous and evoked release of acetylcholine from rat cortical and striatal slices. AB - 1 The effects of two synthetic enkephalins, D-Ala2-Met5-enkephalinamide (DALA) and D-Ala2-MePhe4-Met5-(0)-ol-enkephalin (33,824) were tested on the spontaneous and stimulated release of acetylcholine (ACh) from rat cortical and striatal slices. 2 DALA, but not 33,824, caused a modest increase in spontaneous cortical ACh release. Neither enkephalin affected the release of cortical ACh induced by high potassium (25 mM) or veratridine (20 microM). 3 DALA had no effect on the release of ACh from striatal slices occurring spontaneously or in the presence of potassium (25 mM), veratridine (5 microM) or ouabain (10 microM). The enkephalin 33,824 also had no significant action on the striatal ACh release except that it caused a slight enhancement of veratridine-evoked release. 4 It is suggested that enkephalins have no significant action on release of ACh from the cortex or striatum in vitro. Their action on ACh release from the cortex in vivo, seen in previous studies, may be exerted at a subcortical level. PMID- 7470738 TI - Inhibition of monoamine oxidase by furazolidone in the chicken and the influence of the alimentary flora thereon. AB - 1 The addition of furazolidone to the feed at the therapeutic level (0.04% w/w, 10 days) inhibited monoamine oxidase (MAO) activity by 47 to 72% in chicken duodenal mucosa, heart and brain, but in the liver the enzyme activity was unaffected by the treatment. 2 Furazolidone (200 mg/kg) administered by crop tube inhibited MAO activities in duodenal mucosa, liver, heart and brain. 3 Furazolidone (200 mg/kg) injected intramuscularly did not inhibit MAO activity in the chicken. 4 Pretreatment of the chickens with intramuscular neomycin did not antagonize the inhibition of MAO activity produced by furazolidone (200 mg/kg, crop tube). 5 Pretreatment with neomycin by crop tube to suppress the alimentary flora significantly reduced the effect of furazolidone on MAO activity, suggesting that the drug was transformed by the alimentary flora to an active metabolite which subsequently inhibited MAO activity in other organs. 6 Furazolidone in the feed (0.04% w/w, 10 days) or administered by crop tube (200 mg/kg) had no effect on the activity of aminopyrine demethylase in chicken liver. 7 The activity of aspartate transaminase in plasma was unaffected by the addition of furazolidone to the feed (0.04% w/w, 10 days). PMID- 7470739 TI - The effects of dipyridamole on the guinea-pig ileal longitudinal muscle-myenteric plexus preparation. AB - 1 The longitudinal muscle-myenteric plexus strip prepared from guinea-pig ileum has been used to study the actions of dipyridamole and its interactions with adenine derivatives. 2 Dipyridamole augmented the inhibitory effects of adenine derivatives on the twitch response induced by 0.1 Hz field stimulation of the preparation. This synergistic effect was apparent with relatively low concentrations of dipyridamole (10 to 100 nM) and after short pretreatment times (1 to 2 min) that did not inhibit the twitch response on their own. Appropriate studies suggested the dipyridamole-adenosine synergism followed a pattern of facilitative agonist competition. 3 Dipyridamole did not inhibit either uptake of [3H]-adenosine by the preparation or adenosine deaminase activity under the same conditions that it exhibited synergism with adenosine. 4 Higher concentrations of dipyridamole inhibited the twitch response, mainly by decreasing acetylcholine release but partly by a direct action on smooth muscle. The direct action of dipyridamole on muscle was not synergistic with adenosine. 5 Fluorescence microscopy showed preferential binding of dipyridamole to the myenteric plexus. PMID- 7470740 TI - Binding of [3H]-muscimol, a potent gamma-aminobutyric acid receptor agonist, to membranes of the bovine retina. AB - 1 The binding of [3H]-muscimol, a potent gamma-aminobutyric acid (GABA) receptor agonist, to crude membrane preparations of bovine retina was studied, using a filtration method to isolate membrane-bound ligand. 2 Specific binding was found to be saturable and occurred at two binding sites with affinity constants of 4.3 nM and 38.2 nM. 3 Binding was sodium-independent, enhanced by both freezing and Triton X-100 treatment but abolished with sodium laurylsulphate. 4 The binding sites demonstrated a high degree of pharmacological specificity, GABA being a potent displacer of [3H]-muscimol. 5 A higher degree of [3H]-muscimol binding was associated with subcellular fractions enriched with photoreceptor synaptosomes rather than with fractions enriched with conventional synaptosomes. PMID- 7470741 TI - Uptake of radiocalcium by nerve endings isolated from rat brain: kinetic studies. AB - 1 The uptake of radiocalcium by nerve-ending particles isolated from rat brain was studied in vitro by means of a rapid lanthanum quenching technique. 2 The observed uptake fits a theoretical three-compartment model with two separate uptake phases, a fast, initial phase followed by a late, slow phase. This holds true during control conditions as well as during high-potassium stimulation. 3 The uptake as a function of the external calcium concentration can be described in terms of Michaelis-Menten kinetics during high-potassium stimulation. Under control conditions the fit is clearly applicable but statistically not as good as during potassium stimulation. 4 The affinity for the uptake of calcium remains unchanged under control conditions while during high-potassium stimulation the affinity drastically decreases during the late, slow phase of uptake. 5 During high-potassium stimulation the maximal velocity of calcium uptake is twice that during control conditions. This holds true for both the fast and the slow phases of the uptake. 6 Mg2+ has an inhibitory effect on the uptake, the inhibition being more effective during high-potassium stimulation. Tetrodotoxin has a slight inhibitory effect additional to that extended by Mg2+ during the initial phase of uptake into high potassium stimulated synaptosomes. PMID- 7470742 TI - The size of the hydroxyl group and its contribution to the affinity of atropine for muscarine-sensitive acetylcholine receptors. AB - 1 From measurements of the affinity constants of hydratropyltropine and its methiodide for muscarine-sensitive acetylcholine receptors in the guinea-pig ileum, the increment in log K for the hydroxyl group in atropine is 2.06 and in the methiodide it is 2.16. These effects are slightly bigger than any so far recorded with these receptors. 2 The estimate of the increment in apparent molal volume for the hydroxyl group is 1.1 cm3/mol in atropine and 1.0 cm3/mol in the methobromide. 3 The large effect of the group on affinity may be linked to its small apparent size in water as suggested in the previous paper. PMID- 7470743 TI - Brain and serum calcium concentrations following electroconvulsive shock or bicuculline-induced convulsions in rats. AB - 1 A single electroconvulsive shock (ECS; 125 V, 1 s, 50 Hz) given to rats anaesthetized with halothane produced little change in either serum or regional brain calcium concentrations, compared to rats anaesthetized with halothane, either 5 min or 24 h after the convulsion. Both anaesthetic and ECS-treated rats showed an elevated serum concentration 5 min after the ECS. 2 When 5 ECS were given spread out over 10 days there were no significant calcium concentration changes in either serum or brain except for an increase in the pons/medulla. 3 A single convulsion produced by bicuculline (0.375 mg/kg i.v.) resulted in a marked increase in the calcium concentration in serum, but not brain, 5 min later. Diazepam pretreatment (10 mg/kg i.v.) prevented both the convulsion and the serum calcium change. 4 Results are discussed in relation to clinical data on calcium changes following electroconvulsive therapy (ECT) and the enhanced monoamine mediated behaviours which follow ECS administration to rats. PMID- 7470745 TI - Vasorelaxant action of caroverine fumarate (a quinoxaline derivative), a calcium blocking agent. AB - 1 Caroverine fumarate, 1-(2-diethylaminoethyl)-3-(p-methoxy-benzyl)-1,2-dihydro-2 quinoxalinone fumarate, caused a greater inhibition of the pressor response to KCl (8 x 10(-2) M) than that to noradrenaline (10(-6) M) in the rat hindquarter preparation. 2 In the isolated aorta of the rat, caroverine (up to 10(-6) M) markedly suppressed the contraction caused by KCl (4 x 10(-2) M) (high-K) but had little effect on the contractile response to noradrenaline (10(-6) M) whether added before the spasmogen or in its presence. 3 In the high-K-treated aorta, caroverine shifted the concentration-response curve for external calcium to the right, competitively. The negative logarithm of the affinity (pA2) of caroverine was calculated to be approx. 7. 4 Increased 45Ca uptake of the high-K-treated aorta measured by a modified lanthanum method was inhibited by either caroverine (3 x 10(-6) M) or verapamil (10(-6) M). 5 Concentrations of caroverine and verapamil reducing high-K-induced aortic contraction to 50% of its maximum were 2.4 x 10(-7) and 6.6 x 10(-8) M respectively. 6 Following washout the caroverine induced inhibition of high-K-induced aortic contraction was more rapidly restored than the verapamil-induced inhibition. 7 These results suggest that caroverine fumarate is a specific and readily reversible calcium influx inhibitor in the rat vascular smooth muscle. PMID- 7470744 TI - The interaction of amine local anaesthetics with muscarinic receptors. AB - 1 Amine local anaesthetics inhibited the binding of (-)-[3H]-quinuclidinyl benzilate ((-)-[3H]-QNB) to muscarinic receptors in crude synaptosomal preparations from guinea-pig brain. The order of potency was SKF 525A greater than tetracaine greater than procaine congruent to quinidine greater than procainamide greater than bupivacaine greater than lignocaine greater than prilocaine. 2 The concentration of tetracaine or prilocaine causing 50% inhibition of the receptor-specific binding of [3H]-QNB varied linearly with the concentration of [3H]-QNB present for the range of concentrations of prilocaine used and at lower concentrations of tetracaine, thus providing evidence for a competitive interaction. The affinity constant for tetracaine was 2.6 +/- 0.2 x 10(5) M-1 and that for prilocaine 2.6 +/- 0.8 x 10(3) M-1. At higher concentrations of tetracaine the interaction appears to diverge from simple competitive kinetics. 3 The log dose-response curve for the contractile response of longitudinal muscle strips from guinea-pig intestine to carbachol was shifted in a parallel fashion by low concentrations of tetracaine, but flattened by higher doses. A similar effect was observed for both lignocaine and prilocaine. The affinity constants for tetracaine and prilocaine calculated from the parallel shifts, 1 x 10(5) M-1 and 4 x 10(3) M-1, respectively, were in reasonable accord with the binding data. 4 The curve for the inhibition of [3H]-QNB binding by carbachol was not significantly altered, either in position or shape, in the presence of 1 mM prilocaine. Thus there is no evidence that prilocaine, which increases the affinity of nicotinic acetylcholine receptors for agonists, has any similar effect on agonist binding to muscarinic receptors. PMID- 7470746 TI - Pharmacological study of the anococcygeus muscle of the dog. AB - 1 The response of the dog anococcygeus muscle to field stimulation and to some drugs has been studied. The results are compared with those reported previously in the rat, cat and rabbit. 2 Field stimulation produced frequency-dependent contractions which were inhibited by guanethidine and phentolamine. When the tonus of the muscle was increased with guanethidine, field stimulation always produced frequency-dependent relaxation. The relaxation was not prevented by propranolol. 3 The muscle was contracted by noradrenaline, tyramine, acetylcholine, histamine (H1), 5-hydroxy-tryptamine, prostaglandin E2 and vasopressin. Phentolamine, atropine, promethazine (but not cimetidine) and methysergide inhibited the effect of the respective agonists. 4 After increasing the tonus of the muscle, it was relaxed by low concentrations of isoprenaline. The relaxation was antagonized by propranolol. 5 The response to adenosine triphosphate (ATP) was variable. In some preparations, it relaxed the muscle, in others it contracted the muscle prior to relaxation, in others still it only contracted the muscle. Indomethacin did not prevent ATP-induced contraction. 6 It is concluded that the anococcygeus of the dog, like that of rat, cat and rabbit, has an adrenergic motor innervation and an inhibitory innervation, the transmitter of which is not identified. PMID- 7470747 TI - Mechanism of noradrenaline potentiation by prostaglandin E2 in rat mesenteric artery. AB - 1 Effects of prostaglandins E2 and F2 alpha (PGE2 and PGF2 alpha) on vasoconstrictor responses to noradrenaline (NA) and methoxamine in isolated mesenteric arteries of the rat were investigated. 2 PGE2 and to a lesser extent PGF2 alpha potentiated vasoconstrictor responses to NA and methoxamine. 3 Prior treatment with reserpine increased, and bretylium reduced, the extent of potentiation significantly. 4 NA vasoconstriction persisted for 1 h after Ca2+ was removed from the perfusing Krebs solution. Prostaglandin-induced potentiation was absent in Ca2+-free Krebs, but increased proportionately with increase in external Ca2+ concentration. 5 Vasoconstriction induced by high potassium, was not potentiated by PGE2. 6 It is concluded the PGE2 potentiates NA vasoconstriction by facilitating Ca2+ influx. PMID- 7470748 TI - Evidence for the metabolic activation of non-steroidal antioestrogens: a study of structure-activity relationships. AB - 1 The oestrogenic and antioestrogenic activities of tamoxifen and monohydroxytamoxifen have been compared with those of para-methoxy, -methyl, fluoro, and -chloro tamoxifen in the 3 day immature rat uterine weight test.2 The oestrogenic activity of mestranol, a steroid with low oestrogen receptor binding affinity which is believed to be demethylated to ethinyl oestradiol before exerting its effects, was less potent than ethinyl oestradiol when assayed in the 3 day immature rat uterine weight test. Similarly, para-methoxytamoxifen was less active than monohydroxytamoxifen in oestrogenic and antioestrogenic tests.3 The introduction of a para-methoxy group into tamoxifen did not affect oestrogenic or antioestrogenic activity.4 All the derivatives of tamoxifen were partial oestrogen agonists when compared with oestradiol benzoate in the 3 d immature rat uterine weight test. All test compounds inhibited the uterotrophic activity of oestradiol benzoate (0.16 mug daily) in a dose-related manner. The order of potency was: monohydroxytamoxifen > tamoxifen identical with methoxytamoxifen > p fluoro identical with p-chloro identical with p-methyltamoxifen.5 Tamoxifen was approximately equiactive with its p-methyl, p-fluoro and p-chloro derivatives in the ability to inhibit [(3)H]-oestradiol binding to rat uterine oestrogen receptors in vitro.6 Tamoxifen was approximately equiactive with its p-methyl and p-fluoro derivatives in the ability to inhibit vaginal cornification of ovariectomized rats upon intravaginal administration with oestradiol (3.2 ng total dose).7 Since tamoxifen in vivo was more active as a partial oestrogen agonist and antagonist than the para substituted fluoro, chloro and methyl derivatives that cannot undergo metabolic hydroxylation to monohydroxytamoxifen, whereas the antioestrogenic activity of the compounds upon local application in the vaginal cornification test was equivalent as was their ability to inhibit [(3)H]-oestradiol-17beta binding to the oestrogen receptor in vitro, it is suggested that at low doses; i.e. over the range of the partial agonist dose response curve, the biological activity of tamoxifen is the net result of the activities of the parent compound and its metabolites.8 The results demonstrate that metabolic activation of non-steroidal antioestrogens is only an advantage and not a requirement for antioestrogenic activity. PMID- 7470749 TI - The effect of gamma-aminobutyric acid on the input conductance and membrane potential of Ascaris muscle. AB - 1 Twin intracellular recordings were made from the bag region of Ascaris muscle in order to make conductance measurements. The preparation was bathed in a cool (22 degrees C) Ringer solution to abolish the large spontaneous depolarizing potentials and to improve stability for recording.2 The resting membrane potential was -31 +/- 1 mV, mean +/- s.e. mean (n = 17). The current-voltage plots were linear in the hyperpolarizing direction but showed evidence of delayed rectification during the application of depolarizing currents. The input conductance of the bag was measured from the slope of these plots during the application of hyperpolarizing current. The resting conductance of the bags was 2.4 +/- 0.2 muS, mean +/- s.e. mean (n = 12).3 When the preparation was perfused with gamma-aminobutyric acid (GABA) in concentrations greater than 3 muM, a dose dependent increase in conductance associated with a hyperpolarizing potential was recorded. The log dose-response relationship obtained from 6 preparations was sigmoidal and had an ED(50) of 13 muM.4 When Cl(-) in the Ringer was replaced by SO(4) (2-), the GABA-induced conductance changes decreased and were associated with depolarizing potentials.5 Voltage responses were recorded in the bag region during the iontophoretic application of GABA to different regions of the muscle cell. The largest responses were recorded when GABA was applied to the bag region. Smaller responses were recorded when GABA was applied to the arms and syncytial regions. The responses of the bags were dose-dependent and were antagonized by bath-applied picrotoxin.6 An extrapolation method using 10(-5) M GABA suggested that the GABA reversal potential was about 30 mV more negative than the resting membrane potential. This was confirmed by means of a two microelectrode voltage clamp technique. The reversal potential was thus estimated as -61.2 +/- 2.2 mV mean +/- s.e. mean (n = 27). PMID- 7470750 TI - The interaction of trazodone with rat brain muscarinic cholinoceptors. AB - The muscarinic receptor binding of trazodone, a new nontricyclic antidepressant, was compared with established tricyclic antidepressants. The ability to inhibit the binding of [3H]-quinuclidinyl benzilate in vitro was used for comparing atropine-like effects. Trazodone was found to have essentially no activity at the muscarinic acetylcholine binding site in comparison to the tricyclic antidepressants. PMID- 7470751 TI - Antagonist discrimination between ganglionic and ileal muscarinic receptors. AB - The effects of four antagonists on the depolarization of isolated superior cervical ganglia and the contraction of isolated ileal segments of the rat were compared. pA2 values estimated from Schild plots indicated significantly higher affinities of stercuronium (x100) and pirenzepine (x23) and a significantly lower affinity of 4-diphenylacetoxy-N-methylpiperidine methiodide (x0.39) for the ganglion than for the ileum. The affinities of N-methylscopolamine for the two tissues were not significantly different. It is concluded that the two types of muscarinic receptor are not identical. PMID- 7470752 TI - The influence of hormonal and neuronal factors on rat heart adrenoceptors. AB - 1 The influence of hormonal and neuronal factors on adrenoceptors mediating increased cardiac force and rate of contraction were studied in rat isolated atria. The pharmacological properties of these receptors were deduced from the relative potencies of agonists and from the effects of selective alpha- and beta adrenoceptor antagonists. The numbers and affinities of alpha- and beta adrenoceptors were also determined by radioligand binding to ventricular membrane fragments.2 Hypophysectomy reduced the inotropic potency of isoprenaline and increased the potency of phenylephrine and methoxamine in left atria. The effect of phenylephrine was inhibited by propranolol less effectively and by phentolamine or phenoxybenzamine more effectively in hypophysectomized than in control rats. The difference in block was smaller at low than at high antagonist concentrations. Similar but smaller changes were observed for chronotropic responses of right atria.3 The decreased beta- and increased alpha-receptor response after hypophysectomy was similar to that observed earlier in thyroidectomized rats (Kunos, 1977). These changes developed slowly after hypophysectomy (>2 weeks), they were both reversed within 2 days of thyroxine treatment (0.2 mg/kg daily), but were not affected by cortisone treatment (50 mg/kg every 12 h for 4 days).4 Treatment of hypophysectomized rats for 2 days with thyroxine increased the density of [(3)H]-dihydroalprenolol ([(3)H]-DHA) binding sites from 27.5 +/- 2.7 to 45.5 +/- 5.7 fmol/mg protein and decreased the density of [(3)H]-WB-4101 binding sites from 38.7 +/- 3.1 to 18.7 +/- 2.5 fmol/mg protein. The affinity of either type of binding site for agonists or antagonist was not significantly altered by thyroxine treatment and the sum total of alpha(1)- and beta-receptors remained the same.5 Sympathetic denervation of thyroidectomized rats by 6-hydroxydopamine increased the inotropic potency of isoprenaline and noradrenaline and the blocking effect of propranolol, and decreased the potency of phenylephrine and the blocking effect of phenoxybenzamine to or beyond values observed in euthyroid controls. The density of [(3)H]-DHA binding sites was higher and that of [(3)H]-WB-4101 binding sites was lower in the denervated than in the innervated hypothyroid myocardium. Depletion of endogenous noradrenaline stores by reserpine did not significantly alter the adrenoceptor response pattern of the hypothyroid preparations and did not influence the density or affinity of [(3)H]-DHA and [(3)H]-WB-4101 binding sites.6 These results indicate that thyrotropin or steroids do not contribute to the reciprocal changes in the sensitivity of cardiac alpha(1)- and beta adrenoceptors in altered thyroid states. These thyroid hormone-dependent changes are probably due to a parallel, reciprocal change in the numbers but not the affinities of alpha(1)- and beta-adrenoceptors. Reciprocal regulation of cardiac alpha(1)- and beta-adrenoceptors by thyroid hormones requires intact sympathetic innervation but not the presence of normal stores of the neurotransmitter. PMID- 7470753 TI - The response of the isolated ductus arteriosus to transmural stimulation and drugs. AB - 1 Responses of isolated ductus arteriosus preparations from near term guinea-pigs and lambs to transmural electrical stimulation and drugs were studied in a low oxygen medium (Po(2) 19 to 28 mmHg).2 Acetylcholine and noradrenaline contracted both vessels in a dose-dependent manner, their threshold being between 10(-8) and 10(-7) M. Transmural stimulation (pulse width 0.2 to 0.6 ms, typically 20 Hz) also contracted the vessels.3 Atropine and phentolamine or dibenzyline selectively blocked responses to acetylcholine and noradrenaline, respectively.4 In the guinea-pig ductus, part of the response to transmural stimulation was due to activation of intrinsic adrenergic nerves since the responses were reduced by alpha-adrenoceptor antagonists, bretylium or prior reserpine treatment, but not by atropine. The response of the lamp ductus to transmural stimulation varied greatly in magnitude and was inconsistently affected by alpha-adrenoceptor blocking drugs.5 There was no evidence that transmural stimulation activated cholinergic nerves in either species.6 After inactivation of alpha-adrenoceptors with dibenzyline, noradrenaline caused a beta-adrenoceptor-mediated relaxation. Both this effect and isoprenaline-mediated relaxation were blocked by propranolol. beta-Adrenoceptor activity was more prominent in the ductus of the guinea-pig than of the lamb.7 Raising the Po(2) from 19-28 to 92-98 mmHg increased the response of the guinea-pig ductus to transmural stimulation suggesting that, in this species, physiological elevation of oxygen tension at birth may increase transmitter release from intrinsic adrenergic nerves. Whether this mechanism would contribute to ductus closure remains an open question.8 We postulate that beta-adrenoceptor-mediated relaxation has a role in maintaining ductus patency in the guinea-pig foetus. PMID- 7470754 TI - The effects of time and indomethacin on contractile responses of the guinea-pig gall bladder in vitro. AB - 1. The effects of time and of indomethacin on contractile responses of the guinea pig gall bladder were studied in vitro. 2. The tissues contracted to field stimulation at 5 Hz (in the absence and presence of atropine 10(-6) M). (-) noradrenaline (10(-5) M), acetylcholine (10(-5) M), and adenosine 5'-triphosphate (ATP, 10(-4) M); the magnitude of the contractile responses increased with time. 3. (-)-Isoprenaline 10(-5) M either relaxed (17 of 23 preparations tested) or had no effect on gall bladder strips. 4. The responses of strips of guinea-pig gall bladder to field stimulation at 5 Hz (in the absence or presence of atropine 10( 6) M), (-)-noradrenaline (10(-5) M), and acetylcholine (10(-5) M) obtained 4 h 45 min after setting up the tissue were reduced following incubation with indomethacin (7.9 x 10(-6) M for 1 h). The responses to (-)-isoprenaline (10(-5) M) and to ATP (10(-4) M) were abolished by incubation with indomethacin. 5. These results suggest that, in the guinea-pig gall bladder in vitro, the magnitude of the contractile responses to field stimulation at 5 Hz, (-)-noradrenaline (10(-5) M), and acetylcholine (10(-5) M) and the ability of the tissue to respond to (-) isoprenaline (10(-5) M) and ATP (10(-4) M), may be dependent on the synthesis of a prostaglandin-like substance. PMID- 7470755 TI - The distribution of adrenoceptors and other drug receptors between the two ends of the rat vas deferens as revealed by selective agonists and antagonists. AB - 1. The effects of adrenoceptor agonists and other agonists on the contractile responses of the prostatic and epididymal portions of the rat isolated vas deferens to single pulse field stimulation were investigated. 2. alpha Adrenoceptor agonists produced prejunctional alpha 2-mediated inhibition and post junctional alpha 1-mediated potentiation of the nerve-induced responses. Guanabenz and xylazine produced mainly inhibitory effects, xylazine being 10 times less potent. Clonidine and oxymetazoline produced inhibition with similar potency to guanabenz but at higher concentrations excitatory effects were present, particularly in the epididymal portion. Phenylephrine produced only potentiation of the nerve-induced response in both portions. Potentiation of nerve-induced responses was a more sensitive and quantitative index of excitation than was direct contraction of the tissue. 3. Isoprenaline and salbutamol both gave beta 2-mediated inhibition of the nerve-induced responses in both portions of tissue. At least part of the effect was post-junctional since phenylephrine contractions were inhibited. Isoprenaline also produced a post-junctional alpha 1 mediated excitation. 4. Noradrenaline produced effects qualitatively similar to those of the other alpha-adrenoceptor agonists, inhibition and excitation predominating in the prostatic and epididymal ends respectively. 5. Morphine produced inhibition in the mouse but not in the rat vas deferens. In rat vas, however, enkephalin analogues produced pre-junctional inhibition of responses in both portions which could be partly reversed by naloxone; restoration of the adrenergic component was more complete. Rat anococcygeus showed no equivalent effect. 6. Adenosine 5'-triphosphate (ATP) inhibited nerve-induced responses in each portion with a greater effect on the prostatic portion. PMID- 7470756 TI - Potentiation of the effects of adenosine on isolated cardiac and smooth muscle by diazepam. AB - 1. Adenosine (10(-7) to 3 x 10(-4) M) or 2-chloroadenosine (10(-8) to 10(-5) M) produced concentration-dependent inhibition of the responses of the rat isolated vas deferens to electrical field stimulation. In electrically driven (2 Hz) guinea-pig isolated left atria, adenosine (10(-6) to 10(-3) M) or 2 chloroadenosine (10(-8) to 10(-7) M) produced concentration-dependent decreases in isometric tension. 2. Diazepam (10(-6) and 10(-5) M) had no direct effect per se, but significantly potentiated the inhibitory action of adenosine on both tissues without altering the inhibitory effect of 2-chloroadenosine. 3. The adenosine uptake inhibitors, hydroxynitrobenzylthioguanosine (HNBTG, 10(-5) M) and dipyridamole (10(-5) M) also potentiated the inhibitory actions of adenosine in rat vas deferens, but not hose of 2-chloroadenosine. 4. Following adenosine uptake inhibition in rat vas deferens by HNBTG (10(-5) M), diazepam (10(-5) M) failed to produce any significant further potentiation of the inhibitory action of adenosine. 5. It is concluded that the potentiation of adenosine by diazepam is possibly due to an inhibition of adenosine uptake. PMID- 7470757 TI - Myotonia as a side effect of diuretic action. AB - 1. Commonly used loop diuretics produce side effects in man which are similar to chemically induced myotonia. These diuretics have structural affinity with known myotonic agents. 2. We have observed EMG myotonia in vivo in leg muscles of rats treated with intravenous frusemide. 3. In the presence of several different diuretics, rat isolated diaphragm, soleus and extensor digitorum longus muscles as well as frog sartorius muscles produce typically myotonic contractions with relaxation times up to several seconds. 4. Intracellular recording of action potentials from diuretic-treated muscles reveals long lasting after-discharges following a brief electrical stimulus, again typical of chemically induced myotonia. 5. Having demonstrated a myotonic action of several diuretics we suggest a need for caution in using these drugs in persons with hereditary myotonia and a need to be aware of possible provocation of myotonia in subclinical cases. Myopathies and neuropathies which are known to result from chronic exposure to myotonic agents also need to be considered. 6. In our study, the diuretic, acetazolamide, unmasked subthreshold myotonia. This seems to be at variance with reports of its usefulness in the treatment of myotonia. 7. Diuretics should probably not be employed in the treatment of herbicide intoxication where their myotonic activity would be expected to add to the known myotonic activity of the herbicide. PMID- 7470758 TI - Clonidine distribution in the rat: temporal relationship between tissue levels and blood pressure response. AB - 1. The time course of the distribution of clonidine (20 microgram/kg, i.v.) was determined in the rat by use of a sensitive and specific radioimmunoassay, and was compared with the hypotensive response following this dose. 2. Levels of clonidine were determined in tissues at 2 min, corresponding to the beginning of the hypotensive phase of the drug, and then at 10, 30 and 120 min during recovery of blood pressure to the pre-dose level. The peak tissue concentrations of clonidine were found at 2 min, after which they declined in a mono-exponential manner. The half-lives of clonidine in the various tissues were similar to the half-life of the recovery of blood pressure. 3. Regional variations in clonidine distribution in the brain were not very great; however, the half-life was longer in the corpus striatum and shorter in the cerebellum than in other brain regions. 4. Clonidine concentrations were highest in the kidney and similarly distributed between the cortex and medulla. Concentrations of the drug in other tissues approximated those in brain. 5. Although clonidine is thought to act primarily through the central nervous system, this distribution study shows that at the peak of the hypotensive response less than 2% of the injected dose is present in brain and at least equal concentrations of the drug are found in most peripheral tissues. Thus the possibility of peripheral mechanisms contributing to the hypotensive effect cannot be dismissed. PMID- 7470759 TI - A comparison of the ability of two angiotensin II receptor blocking drugs, 1-Sar; 8-Ala angiotensin II and 1-Sar, 8-Ile angiotensin II, to modify the regulation of glomerular filtration rate in the cat. AB - 1 Modest stimulation of the renal nerves in the anaesthetized unilaterally nephrectomized cat resulted in a 15% fall in renal blood flow, no change in glomerular filtration rate and significant falls in both the absolute and fractional rates of sodium excretion.2 The haemodynamic responses to nerve stimulation were not modified by angiotensin II blockade with 1-Sar, 8-Ala angiotensin II although the fall in absolute, but not fractional sodium excretion was significantly larger. In contrast, stimulation of renal nerves following administration of 1-Sar, 8-Ileangiotensin II caused a significant fall in glomerular filtration rate. The reductions in both absolute and fractional sodium were of the same magnitude as in the absence of drug.3 Both renal blood flow and glomerular filtration rate were autoregulated during the reduction of renal perfusion pressure and this was associated with reductions in both absolute and fractional sodium excretions.4 In the presence of 1-Sar, 8-Ala angiotensin II, the haemodynamic and sodium excretory responses to reductions in renal perfusion pressure were not significantly different from those recorded in the absence of drug. However, following administration of 1-Sar, 8-Ile angiotensin II, renal blood flow but not glomerular filtration rate, was autoregulated during reduction in renal perfusion pressure. The falls in absolute and fractional sodium excretions caused by this manoeuvre were of similar magnitude to those obtained in the absence of drug.5 The results obtained using the 1-Sar, 8-Ile angiotensin II are consistent with angiotensin II having an important intra-renal site of action to regulate glomerular filtration rate, possibly via an action at the efferent arteriole. Administration of 1-Sar, 8-Ala angiotensin II was without effect on the regulation of renal haemodynamics which it is suggested reflects a limitation in the use of this particular compound as an intrarenal angiotensin II antagonist. PMID- 7470760 TI - Prejunctional actions of piribedil on the isolated kidney of the rabbit: comparison with apomorphine. AB - 1 The effects of piribedil on contractile responses and noradrenaline release evoked by sympathetic nerve stimulation have been studied in the isolated kidney of the rabbit. These effects were compared to those of apomorphine.2 Electrical stimulation (2, 5 and 10 Hz) of sympathetic renal nerves produced frequency dependent increases in perfusion pressure and noradrenaline release. Piribedil did not affect (0.1 mug/min) or diminished (1 and 10 mug/min) the stimulation evoked increase in perfusion pressure, and increased noradrenaline release in a dose-dependent manner.3 Increases in renal perfusion pressure and noradrenaline release induced by electrical stimulation were decreased by apomorphine (0.1 and 1 mug/min). These inhibitory effects were more marked at low frequencies of stimulation and were prevented by haloperidol (0.2 mumol/1).4 Piribedil (0.1 and 1 mug/min) and apomorphine (0.1, 1 and 10 mug/min) did not affect the increases in renal perfusion pressure elicited by exogenously administered noradrenaline, but piribedil (10 mug/min) diminished them.5 In the presence of desipramine (0.5 mumol/l), piribedil (0.1, 1 and 10 mug/min) produced a dosedependent inhibition of the increases in renal perfusion pressure and noradrenaline release evoked by sympathetic nerve stimulation; the inhibitory effect of piribedil was more marked at low frequencies of stimulation and was prevented by haloperidol.6 Piribedil increased the resting release of noradrenaline from the rabbit kidney, in contrast to apomorphine, which was without effect.7 It is suggested that piribedil has a complex effect on sympathetic transmission. This drug exhibits an ;amphetamine-like' action, causing noradrenaline release from its postganglionic stores. This releasing effect masks an action on prejunctional inhibitory dopamine receptors. In addition, at high doses, piribedil exhibits a marked action on postjunctional sites, since it reduces the vasoconstrictor effect of exogenous noradrenaline. PMID- 7470761 TI - Evidence that cell bodies of non-cholinergic, excitatory neurones which supply the smooth muscle of the chicken rectum are located in the ganglia of Remak's nerve. AB - 1. A pharmacological investigation of the distribution of non-cholinergic excitatory nerve cell bodies was performed on the chicken's isolated perfused rectum with attached Remak's nerve supply. 2. Electrical stimulation of Remak's nerve trunk produced a contraction and a discharge of action potentials in the nerve branches which supply the smooth muscle of the rectum. Both responses were virtually blocked by hexamethonium when applied via the caudal mesenteric artery. 3. The contractile effect following stimulation of the nerve trunk was inhibited more effectively by hexamethonium when application was restricted to the trunk rather than to the intestine. 4. The contractile effect of stimulating the nerve branches was unaffected by hexamethonium. 5. It is concluded that ganglionic transmission, which is mediated by nicotinic receptors, occurs in the ganglia of Remak's nerve but not in ganglia of the enteric plexuses. Therefore, cell bodies of the postganglionic neurones which are considered to be non-cholinergic are located in Remak's nerve ganglia. PMID- 7470762 TI - Accumulation and overflow of 3H following incubation of the guinea-pig gall bladder with [3H]-noradrenaline. AB - 1 Strips of guinea-pig gall bladder readily accumulate (3)H following incubation in the presence of 5 x 10(-8) M (-)-[(3)H]-noradrenaline. This accumulation was reduced by lowering the incubation temperature (from 37 degrees to 23 degrees C), by cocaine (10(-6) M), by nortriptyline (10(-8), 10(-6) and 10(-4) M) and following incubation of the tissues with 6-hydroxydopamine (10(-3) M for 3 h). At 10(-6) M, (-)-noradrenaline and (-)-adrenaline, but not (-)-isoprenaline, inhibited the accumulation of (3)H.2 Following preloading of strips of guinea-pig gall bladder with 3.6 x 10(-7) M (-)-[(3)H]-noradrenaline for 1 h, the spontaneous overflow of (3)H was observed. Cocaine (10(-4) M), nortriptyline (10( 6) M), (-)-isoprenaline (10(-5) M), acetylcholine (10(-5) M) and adenosine 5' triphosphate (ATP, 10(-4) M) had no effect on the spontaneous overflow of (3)H. KCl (10(-1) M), (-)-noradrenaline (10(-5) M), (-)-adrenaline (10(-5) M), and tyramine (10(-5) M) increased the overflow of (3)H. These results illustrate similar characteristics of the guinea-pig gall bladder to other noradrenergically innervated tissues in accumulating and releasing (3)H following incubation in the presence of [(3)H]-noradrenaline.3 Following incubation in the presence of 3.6 x 10(-7) M (-)-[(3)H]-noradrenaline, field stimulation, at 5 Hz, of strips of gall bladder, in the absence or presence of 10(-6) M atropine, increased the overflow of (3)H and, simultaneously, induced contractions. The contractile responses to 5 Hz were smaller in the presence than in the absence of 10(-4) M lignocaine. Lignocaine (10(-4) M) reduced the overflow of (3)H evoked by field stimulation at 5 Hz. It is suggested that the contractile responses to 5 Hz are due to nerve stimulation and that the increased overflow of (3)H is due to the stimulation of noradrenergic nerves.4 The overflow of (3)H evoked by field stimulation at 5 Hz was unaltered and increased by propranolol (10(-6) M) and phentolamine (10(-6) M), respectively. Clonidine (5 x 10(-5) M) had no effect in the absence but reduced the amount of (3)H which overflowed in response to field stimulation at 5 Hz in the presence of 10(-6) M atropine. The contractile responses to field stimulation at 5 Hz were reduced by phentolamine (10(-6) M) or clonidine (5 x 10( 6), 10(-5) and 5 x 10(-5) M) whether or not atropine (10(-6) M) was present. These results illustrate the presence of postsynaptic alpha-adrenoceptors and suggest the presence of presynaptic alpha-adrenoceptors in the gall bladder of the guinea-pig. PMID- 7470764 TI - Prophylactic lignocaine and early post-coronary artery occlusion dysrhythmias in anaesthetized greyhounds. AB - 1. Lignocaine (1 mg kg-1 min-1 infused intravenously for 30 min) greatly reduced the incidence of ventricular ectopic beats that resulted from acute coronary artery ligation in anaesthetized greyhound dogs. However, the incidence of ventricular fibrillation was only slightly reduced by this treatment which caused significant myocardial depression. 2. There is no good evidence from this study that lignocaine is a particularly effective prophylactic in acute myocardial infarction. PMID- 7470763 TI - An increased calcium sensitivity of mesenteric resistance vessels in young and adult spontaneously hypertensive rats. AB - 1. We have measured the calcium sensitivity in response to noradrenaline stimulation and potassium depolarization of isolated segments of 100 to 200 micrometer mesenteric resistance vessels from spontaneously hypertensive (SHR) and control Wistar-Kyoto rats. The rats were either young (4 wk) or adult (4 months), that is of ages before or after the SHRs had developed elevated blood pressure. Experiments were performed under conditions in which the effect of noradrenaline uptake by, and the potassium-induced noradrenaline release from, the nerve terminals in the vessel walls was eliminated. 2. The response of the SHR and WKY vessels to noradrenaline under conditions where only the extracellular calcium appeared to have been removed was similar. When subsequently stimulated maximally by noradrenaline, the calcium-sensitivity of the SHR vessels (Ca-ED50 approximately or equal to 0.1 mM) was greater than that of the WKY vessels (Ca-ED50 approximately or equal to 0.2 mM). When depolarized by potassium, all vessels were less sensitive to calcium and there was little difference in the calcium sensitivity of SHR and WKY vessels in either age group (Ca-ED50 approximately or equal to 0.8 mM). 3. The results suggest that whereas the potassium (potential)-dependent calcium permeability of the SHR smooth muscle cell membrane is normal, the noradrenaline-induced calcium permeability is abnormally high. The presence of this abnormality in the vessels from the young SHRs suggests that it may be a factor involved in the aetiology of hypertension in the SHR. PMID- 7470765 TI - Evaluation of antagonism of aconitine-induced dysrhythmias in mice as a method of detecting and assessing antidysrhythmic activity. AB - 1 Antagonism of aconitine-induced dysrhythmias in mice as a method of detecting and assessing antidysrhythmia activity was evaluated. 2 Aconitine-induced dysrhythmias in mice appear to be selectively sensitive to antidysrhythmia agents (administered intraperitoneally) which reduce the inward sodium current in cardiac cells. 3 Antidysrhythmic agents whose mechanism of action is thought to depend on beta-adrenoceptor blockade, prolongation of cardiac monophasic action potentials or calcium antagonism are ineffective in delaying the onset of aconitine-induced dysrhythmias in mice. The inactive drugs were practolol, sotalol, bretylium, amiodarone and verapamil. 4 Comparisons of anti-dysrhythmic activities of test drugs should be based on more than one ED value and should take account of efficacy as well as potency. 5 The mouse aconitine test is a useful and rapid method of evaluating oral antidysrhythmic activity in terms of potency, efficacy and duration of action. 6 With respect to potency, efficacy, oral activity, duration of action and safety, 3 alpha-amino-5 alpha-androstan-2 beta-ol-17-one hydrochloride (Org 6001) offered the most satisfactory overall profile of the active drugs tested (Org 6001, aprindine, quinidine, disopyramide, lignocaine, mexiletine, procainamide and propranolol). PMID- 7470766 TI - Investigation of the antifibrillatory activity of some anticonvulsant gamma aminobutyric acid-transaminase inhibitors in the rabbit isolated heart: comparison with phenytoin and mexiletine. AB - 1 The electrical stimulation and ventricular fibrillation thresholds were measured in the Langendorff-perfused rabbit heart by application of rectangular impulses of 3 ms duration and increasing current at frequencies of 4 and 20 Hz respectively. 2 Perfusion with either phenytoin or mexiletine produced significant dose-dependent increases in both thresholds. 3 Perfusion with four anticonvulsant inhibitors of gamma-aminobutyric acid (GABA)-transaminase, aminooxyacetic acid, gamma-acetylenic GABA, gamma-vinyl GABA and sodium valproate, up to a concentration of 20 microgram/ml, had no significant effect on either threshold. 4 The results suggest that these compounds, unlike phenytoin and mexiletine, are devoid of antifibrillatory activity. PMID- 7470767 TI - Childhood autism and social class: a question of selection? AB - Children with typical autism, other early childhood psychoses and severe mental retardation without autistic behaviour were identified in an epidemiological study in an area of South East London. The social class distribution of their fathers was examined and no significant differences were found between the groups, nor in a comparison with the general population of the area. Fathers of children with autism and related conditions referred to an out-patient clinic with a special interest in autism, mostly at their own request, and fathers joining the National Society for Autistic Children, were of higher social class than both the average for England and Wales and the fathers of the study children. Joining the NSAC during its early years, and keeping up membership were also linked with higher social class. The findings supported the view that reports of a social class bias in autism may be explained by factors affecting referral and diagnosis. PMID- 7470768 TI - Differential use of psychiatric services by West Indians, West Africans and English in London. AB - Two hundred and ninety West Indian and 73 West African patients were identified and matched against 204 and 53 English patients respectively. Significantly more migrant patients were: not referred to hospital by their GPs, showed disturbed behaviour prior to psychiatric contact, and were admitted formally. Studying consecutive attendances or admissions to one hospital may lead to an unrepresentative sample. PMID- 7470769 TI - The peripheral anticholinergic activity of tricyclic antidepressants: comparison of amitriptyline and desipramine in human volunteers. AB - The effects of three single oral doses (25 mg, 50 mg and 100 mg) of amitriptyline and desipramine, and of placebo, were compared on a range of cholinergic functions (resting pupil diameter, pilocarpine-evoked miosis, baseline-sweating, carbachol-evoked sweating, salivation, heart rate) in eight healthy volunteers. Three measures (pilocarpine-evoked miosis, carbachol-evoked sweating and salivation) reflected the antimuscarinic property of the antidepressants; in two tests (pilocarpine-evoked miosis and salivation) amitriptyline appeared to be more potent than desipramine. Resting pupil diameter was not affected by amitriptyline, whereas desipramine caused mydriasis, indicating that pupil size is not a reliable measure of anticholinergic activity in the case of drugs which also affect adrenergic mechanisms. Baseline-sweating and heart rate were not affected by the antidepressants. PMID- 7470770 TI - Long-term monitoring of tricyclic antidepressant plasma concentrations. AB - Plasma levels of antidepressant drugs were measured in 17 depressed patients at roughly monthly intervals over follow-up periods of three to 26 months. Good results in seven were associated with small fluctuations in level from visit to visit, while big fluctuations were associated with poor outcome. Such fluctuations were probably from poor compliance or drug interaction from self medication. Severe side effects and cardiotoxicity were associated with high levels. PMID- 7470771 TI - Lithium therapy in aggressive mentally subnormal patients. AB - In a series of fifteen aggressive mentally subnormal patients treated with lithium carbonate, there was an improvement in aggressive behaviour in eleven cases, three patients showed no change and one patient became worse during lithium treatment. The response to lithium therapy was assessed by clinical observation and retrospective analysis of daily ward reports. Treatment had to be discontinued in one patient who developed tardive dyskinesia, but otherwise side effects were not a problem. PMID- 7470772 TI - Some measures of goal attainment in focussed integrated active psychotherapy: a study of fifteen cases. AB - The psychotherapeutic treatment of 15 patients is described. The goals were defined in terms of target problems and target dilemmas, traps and snags, and the outcome was assessed by rating these or with a symptom questionnaire and a dyad repertory grid before and after treatment. Patients who rated dilemmas as resolved showed appropriate changes in construct correlations on the dyad grid. PMID- 7470773 TI - Immediate psychological help for self-harmers. PMID- 7470775 TI - The disabilities of chronic schizophrenia. PMID- 7470774 TI - The abuse of anti-cholinergic drugs in adolescents. PMID- 7470776 TI - Serum immunoglobulins in dementia. PMID- 7470777 TI - Sex and crime: the results of a self-report study. PMID- 7470779 TI - Mortality from cancer and all causes among British radiologists. AB - The mortality of men who joined a British radiological society between 1897 and 1954 has been compared with that of (i) all men in England and Wales, (ii) men in social class 1, and (iii) male medical practitioners. Radiologists who entered the profession before 1921 suffered a death rate from cancer 75% higher than that of medical practitioners. Among these men there was a statistically significant excess of deaths from cancers of the pancreas (6 against 1.9 expected), lung (8 against 3.7), and skin (6 against 0.8), and from leukaemia (4 against 0.7). There were 72 deaths from cancer among men who entered the study after 1920 and 68.6 deaths were expected, based upon rates among medical practitioners. For no individual cancer site did the observed number of deaths exceed the expected number. There was some evidence however, that the ratio of observed to expected cancer increased with the duration of time that men were included in the study. Among those followed for more than 30 years there were 30 deaths against 22.1 expected. It is not possible to make a close estimate of the dose of radiation received by the men in this study, but those who entered between 1920 and 1945 could have received an accumulated whole-body dose of the order of 1-5 Gy (100 to 500 rad). For all non-cancer causes of death combined, the death rate among radiologists is lower than that among all men in England and Wales, men in social class 1, and male medical practitioners. The data offer no support for the concept of a non-specific aging effect of radiation. PMID- 7470778 TI - Radiation hazards: 25 years of collaborative research. Sylvanus Thompson memorial lecture, April 1980. PMID- 7470780 TI - Clinical relevance of radiological abnormalities of the craniovertebral junction. AB - Clinical review of a large series of lesions of the craniovertebral angle revealed that no abnormality could be shown by radiological means in 30-50% of cases. Conversely, 30-50% of radiologically demonstrable skeletal disorders at the craniovertebral angle were asymptomatic. PMID- 7470781 TI - Amipaque in coronary angiography and left ventriculography. AB - Amipaque (metrizamide) has been directly compared with Urografin and Triosil in coronary angiography and left ventriculography. Amipaque caused significantly less T-wave deflection than either medium when injected into the coronary arteries, and there was a tendency to less reduction in heart rate. Subjective discomfort was significantly reduced with Amipaque, particularly in left ventriculography. Amipaque was also directly compared in two concentrations, 370 and 300 mg iodine/ml. There was a slight tendency to less ECG change and subjective discomfort with the lower concentration, and there was no significant reduction in image quality. The relatively high cost of Amipaque and the necessity of prior mixing make its routine use in adult cardiology unlikely, but it should be valuable in certain cases when minimum toxicity or reduced discomfort are of special importance. A relatively low iodine concentration, for example 300 mg iodine/ml, is sufficient. PMID- 7470782 TI - Radionuclide uptake during the evolution of experimental acute osteomyelitis. AB - The findings of technetium 99m MDP and gallium 67 citrate scintigraphy are described in the first four days following experimental acute osteomyelitis of the tibia of rabbits. On the day of inoculation, defects in activity are observed in both the perfusion, and bone phases of technetium scintigraphy. Similar defects are achieved by saline injection. On subsequent days technetium scintigraphy reveals an increased activity in the perfusion stage but a persisting defect of activity in the bone phase. Gallium activity is increased throughout the period of the study in inoculated animals but not in those injected with saline. It is concluded that 99Tcm MDP, in its perfusion phase, and 67Ga citrate are highly sensitive indicators of bone inflammation and that early osteomyelitis is characterized by increased activity with these agents together with a technetium bone scan defect. PMID- 7470783 TI - Mammographic practice and dosimetry in Britain. AB - A survey of mammographic techniques and dosimetry in 61 NHS centres in Britain has been completed. Physical parameters measured at each centre included tube output, half-value thickness, peak kilovoltage, surface dose and percentage depth dose. Ionisation and thermoluminescence dosimetry were employed and image quality was assessed subjectively using a test phantom. Absorbed doses are given at the plane surface of an average-sized, 5 cm thick homogenous breast composed of 50% fat and 50% water. The doses fall naturally into the four combinations of molybdenum (Mo) and tungsten (W) targets and film (F) and xerox (X) receptors. The Mo/X group produced the best images of the phantom but also the highest mean dose of 3.26 cGy (rad) for a cranio-caudad (c-c) view and its use does not seem justified for asymptomatic screening. The Mo/Lo-Dose or MIN-R film combination gave a mean dose of 0.95 cGy(rad) for a c-c view whereas the W/X group gave 0.89 cGy(rad) and superior image quality. The W/F group gave the poorest image quality and its continued use for mammography is questionable. PMID- 7470784 TI - Evaluating performance characteristics in computerized tomography. AB - The important radiographic and dosimetric performance parameters associated with computerized tomography (CT) are discussed together with the practical problems encountered when they are being evaluated. The parameters include alignment, slice thickness, noise, low-contrast detectability, spatial resolution, modulation transfer function and patient dose. A set of phantoms for determining these performance characteristics are described. The phantoms are of modular construction permitting measurement of the performance parameters at various sites within a number of 6 cm thick, regular-geometry, and realistic body sections. The regular-geometry sections include 20 cm diameter (skull size) and 33 cm diameter (abdomen size) sections. The realistic anthropomorphic sections include the skull, abdomen and thorax. PMID- 7470785 TI - A survey of 29 EMI CT machines in Britain. AB - During 1978/79 a survey was made of the performance characteristics of 29 EMI brain and body CT scanners in Britain. Comparative data on such performance parameters as noise, uniformity of CT numbers, low-contrast detectability, resolution, modulation transfer function (MTF) and effective photon energy were measured using a set of regular-geometry and anthropomorphic phantoms. In addition, dose profiles were obtained for a single slice using TLD and film. The results of the survey are presented; the dependence of the performance characteristics on the types of machines surveyed and the operating modes employed is discussed. PMID- 7470786 TI - Hydatid disease: the value of whole-body screening by scintigraphy with technetium-99m-labelled red blood cells. AB - Whole-body scintigraphy with 99Tcm-labelled red blood cells was performed in eight patients with hydatid disease. There was good correlation between these studies and other diagnostic procedures which included radiocolloid liver scintigraphy, liver ultrasound, body computed tomography and chest radiography. Whole-body scintigraphy with 99Tcm-labelled red blood cells appears suitable for screening patients with hydatid disease, many of whom have multiple organ involvement. PMID- 7470787 TI - Effect of hyperthermia on the hypoxic fraction in an experimental mammary carcinoma in vivo. AB - The influence of hyperthermia on the fraction of clonogenic hypoxic cells in a CaH mammary carcinoma was investigated using the TCD50 values for radiation treatment under normal or clamped conditions. A one-hour heat treatment at 41.5 and 42.5 degree C given four hours after radiation reduced the hypoxic fraction to 2.3 X 10(-1), 7.4 X 10(-3) and 6.5 X 10(-3) respectively, when compared with the content of hypoxic cells in non-heated tumours. The results indicate that moderate hyperthermia is able to destroy selectively a large proportion of the fraction of radioresistant hypoxic cells in a solid tumour. This effect is probably due to the increased heat sensitivity of cells in a chronically hypoxic and nutritionally deprived acidic environment. PMID- 7470789 TI - A phantom for assessing the resolution at depth of a gamma camera system. PMID- 7470788 TI - Effect of X irradiation on the rectum of the rat. AB - It is demonstrated how a definite part of the rectum of the female Wistar rat can be selectively irradiated without surgery. The effect of X irradiation on the rectum is in the acute phase (second week after irradiation) a haemorrhagic radiation proctitis and in the chronic phase a rectal obstruction. The dose effect relationship using rectal obstruction as the end point is a sigmoid curve with a ED10 of 17.50 Gy, ED50 of 21.5 Gy and ED90 of 27.5 Gy. Latency time decreases exponentially with increasing dose and is 150 days at the ED50. Histological findings show mucosal glands and cysts within the submucosa and a severe submucosal radiation within the submucosa and a severe submucosal radiation fibrosis which results in the rectum becoming a rigid and non-elastic tube which leads to a functional block in transport of faeces. PMID- 7470790 TI - Personalized fixation using a vacuum consolidation technique. PMID- 7470791 TI - Iodine 131 ablation therapy for a patient on maintenance haemodialysis. PMID- 7470792 TI - Intermittent cyanotic attacks in an infant-an unusual presentation of a congenital anterior diaphragmatic (Morgani) hernia. PMID- 7470793 TI - Computed tomography in diabetes insipidus: posterior empty sella. PMID- 7470794 TI - Pneumatosis intestinalis in the small bowel of an adult-a radiological sign of a serious post-operative complication. PMID- 7470795 TI - Use of diagnostic ultrasound to measure the lumbar spine canal. PMID- 7470796 TI - Gold grain dosimetry. PMID- 7470797 TI - Megacalices--how broad a spectrum? AB - Ten patients with a suspected diagnosis of megacalices have had pressure-flow studies to confirm or exclude obstruction. Eight had no evidence of obstruction and amongst these there were 2 patients with a non-obstructed megaureter. Two patients showed evidence of obstruction and remain a dilemma from a management viewpoint. If "megacalices" is used simply as a descriptive term, then the spectrum of disease is wider than previously suspected. PMID- 7470798 TI - Papillary adenocarcinomatous degeneration occurring in a multilocular renal cyst. AB - A multilocular cyst of the kidney in an adult with malignant papillary adenocarcinomatous transformation is described. The histogenesis and prognosis of multilocular cysts are discussed with special reference to their relationship with Wilms' tumour. PMID- 7470799 TI - Use of the surgical staple for bowel anastomoses in urology. AB - The use of the Autosuture stapling devices for performing enteric anastomoses is described. Such anastomoses can be performed more quickly and more uniformly than sutured ones, with few complications. PMID- 7470800 TI - Incontinence following transurethral resection of the bladder neck in the female. AB - Incontinence is an unwelcome complication of transurethral resection (TUR) of the bladder neck in the female. A computerised analysis of 238 procedures performed during the period 1940 to 1972 was carried out to assess the morbidity due to this complication and to evaluate possible predisposing factors. Initially 15.2% of patients were incontinent but 70% of these soon became continent without further treatment. The remaining 4.7% were treated by surgical or other measures. The incidence of incontinence was unrelated to the amount of tissue removed or to the number of operations, but was significantly lower when the operation was done for polypi of the bladder neck and significantly higher in patients with a cystocele or urethrocele. It seems possible to reduce the incidence of incontinence by careful selection of cases. PMID- 7470801 TI - Penile injuries with particular reference to injury to the erectile tissue. AB - A retrospective study of 56 patients with penile injuries showed that 28 injuries were sustained during coitus and a further 12 in other sexual practices. Four different types of injury to the erectile mechanism are described and the management of each discussed. A modified phalloplasty technique is described for patients who have sustained a complete loss of the phallus. PMID- 7470802 TI - The intravenous urogram in acute epididymo-orchitis. AB - In a series of patients presenting with acute epididymo-orchitis, the abnormalities detected on intravenous urography were analysed. Urography is recommended in patients aged over 50 and in pre-pubertal boys, but it did not prove useful in adult patients under 50 years of age without other symptoms. PMID- 7470803 TI - Scrotal lipomatosis. AB - The anatomy of scrotal fat was studied histologically and by dissection in 28 normal cadavers and 44 idiopathic infertile subjects. Two fat patterns were described: normal and infertile. In the normal pattern, a small posterior extratunicary pad of fat was constantly encountered. Intratunicary fat occurred as small granules between the cord veins. Thirty-eight of the 44 idiopathic infertile subjects had scrotal lipomatosis, of which 2 types were recognised: extratunicary and intratunicary. The latter showed 2 patterns: diffuse and lobular. The diffuse pattern occurred in obese subjects and those of normal build, and the fat was firmly adherent to the cord veins. The lobular pattern occurred exclusively in the obese, and the lobules were loosely connected to the cord. The anatomical features of the normal fat pattern in relation to maximal testicular thermoregulatory efficiency were discussed. The role of scrotal lipomatosis in infertility was clarified and the relationship of scrotal lipomatosis to obesity presented. PMID- 7470804 TI - Lipectomy in the treatment of scrotal lipomatosis. AB - Scrotal lipectomy was performed in 102 infertile patients with scrotal lipomatosis; suprapubic lipectomy was included in 22 patients who had redundant suprapubic fat. Improvement in semen quality was obtained in 64.7% and pregnancy occurred in 19.6%. Statistically, the improvement was significant in both the sperm count and motility of the total series and only in the sperm count in the cases which produced pregnancy. The technique is described and the factors to which the results may be attributed are discussed. PMID- 7470805 TI - Urolithiasis in the Sudan. Studies on a stone-prone and a stone-free population. AB - Biochemical studies were performed on 80 adult males with renal stones and 24 controls from Northern Sudan. Similar studies were performed on 44 controls from Southern Sudan, where urinary stones rarely occur. No significant differences between stone formers and controls from Northern Sudan were found regarding serum uric acid, phosphate, alkaline phosphatase, total protein, albumen, daily urine calcium or uric acid. Similarly, no significant difference was found between 2 groups in the incidence of hypercalciuria, hyperuricaemia of hyperuricosuria. The serum calcium was significantly higher in stone formers. The mean serum and daily urine calcium were significantly lower, and the mean serum and daily urine uric acid were significantly higher in controls from Southern Sudan compared to either stone formers or controls from Northern Sudan. The mean daily urine volume was significantly higher in Southern controls compared to Northern controls. The increased prevalence of stone formers in Northern Sudan appears to result from the higher calcium output in the urine and the lower urine volume in the North compared with the South. PMID- 7470806 TI - Ruptured ureter in pregnancy--a unique case? PMID- 7470807 TI - Chronic inflammation of the ureter with urothelial ingrowth simulating inverted papilloma. PMID- 7470808 TI - Bilateral xanthogranulomatous pyelonephritis. PMID- 7470809 TI - Ruptured renal carcinoma in a child. PMID- 7470810 TI - Acute small bowel obstruction 20 years after reimplantation of the ureter. PMID- 7470811 TI - A transurethral biopsy punch for the prostate gland. PMID- 7470812 TI - Lymphoscintigraphy cannot diagnose breast cancer. AB - Axillary lymphoscintigraphy was performed by the technique of Agwunobi and Boak (1) in 68 patients with operable cancer of the breast and 11 patients with benign disease. Contrary to their previous report, scans were found to be indistinguishable from normal in the majority of breast cancer patients with interpretable scans, and no pattern of uptake was found which was typical of carcinoma in the corresponding breast. PMID- 7470815 TI - The effects of caerulein and a meal stimulus on gallbladder intraluminal pressure and bile flow in the pig. AB - The intraluminal gallbladder pressure has been measured in a conscious unsedated pig model by the use of a radiotelemetry pressure capsule. A loop fistula was incorporated to measure bile output simultaneously. Mean fasting intraluminal gallbladder pressure was found to be 15 +/- 4 mmHg above atmospheric pressure, and mean fasting bile output 7.5 +/- 0.8 ml/10 min. Following intravenous caerulein pressure peaked at 2 min with a mean rise of 17 +/- 3 mmHg above basal (P less than 0.005). Bile output was increased for 30 min following caerulein with a total mean increase for this period of 13.5 +/- 2.2 ml (P less than 0.005). Gallbladder pressure rose significantly 5 min after commencing a meal by 6.0 +/- 1.9 mmHg (P less than 0.025) reaching a peak pressure 65 min after the meal with a mean rise of 14.8 +/- 2.3 mmHg (P less than 0.005). Bile output was significantly increased for 50 min following the ingestion of food. The total increments above basal values for bile output being 15.9 +/- 2.7 ml (P less than 0.005) in the first 30 min post-prandially and 1.6 +/- 2.4 ml (P less than 0.005) in the 30--60 min period. This model re-emphasizes the rapidity of the hormonal response and may be valuable for further studies on the physiology of post prandial gallbladder emptying. PMID- 7470814 TI - 111Indium-labelled leucocyte scintigraphy in the diagnosis of inflammatory disease--first results. AB - In 61 patients, 66 scans were performed with 111In-labelled autologous leucocytes to evaluate this method in the diagnosis of inflammatory disease. All scintigraphic results were compared with clinical, operative or postmortem findings. In the first 20 examinations in 19 patients, 2 scintigrams were true positive, 9 true negative one of false positive, whereas 9 scintigrams had to be considered as false negative. These false negative results were probably due to loss of viability of the labelled cells. A modified labelling technique, in which a much lower concentration of oxine was used, was employed for a further 46 examinations in 42 patients. Twenty-four scintigrams were true positive and 22 true negative, while no false positive or false negative results were observed. These results suggest that this modified technique is useful in the diagnosis of inflammatory disease. PMID- 7470816 TI - Evaluation of intended radical surgery in carcinoma of the gallbladder. AB - The records of 44 consecutive patients with carcinoma of the gallbladder from 1968 to 1977 were reviewed. All patients were investigated with a view to resection of the tumour and this was carried out in 10 patients, all macroscopic tumour being excised in 6. The basic operation was cholecystectomy, but liver resection (4 patients), liver resection and colectomy (3 patients) and pancreaticoduodenectomy (1 patient) were also carried out. Of these 10 patients, 3 died in hospital, while of the 34 patents in whom a palliative procedure was possible, 15 died during their hospital treatment. The median survival times were 4 months and 3 months respectively. Only one patient in this series is alive and symptom-free at 36 months. It is concluded that radical surgery, of itself, does not confer any advantage to the group of patients with carcinoma of the gallbladder. Alternative strategies for diagnosis and treatment of this condition are discussed. PMID- 7470813 TI - Scintiscanning of the breast with 99Tcm-diethylene triamine penta-acetic acid--a prospective blind evaluation. AB - In this blind prospective study of breast scintiscanning with 99Tcm-diethylene triamine penta-acetic acid (DTPA) in 67 female patients presenting at a primary breast referral clinic, we have observed that this investigation has a high diagnostic accuracy in distinguishing benign from malignant lesions of the breast. The predictive value of the test when positive in establishing the presence of cancer is 94 per cent with a false negative rate of 6 per cent. The predictive value of the test when negative in excluding cancer in 97 per cent with a false positive rate of 3 per cent. Initial studies also indicate the potential value of DTPA breast scintiscanning in delineating the activity of painful benign lesions of the breast. PMID- 7470817 TI - Carcinoma of the gallbladder. AB - A retrospective study of 100 proved cases of primary carcinoma of the gallbladder admitted to the University Surgical Unit, University of Hong Kong, over a 20-year period was undertaken. The female to male ratio was 1.4 to 1. The peak incidence was in the seventh decade for the females and the sixth decade for the males. Preoperative diagnosis was made in 10 patients. "Curative" cholecystectomy was performed in 20 patients with a 5-year survival rate of 10 per cent. Palliative procedures were performed in 44 patients with a median survival of 8 weeks, which was not significantly different from that in the 30 patients who were not operated upon or had laparotomy and biopsy only. Radical resection was carried out in 6 patients, all of whom died in hospital. Gallstones were found in only 26 patients. The gallbladders of 3 patients had associated benign tumours, one of which had malignant transformation at multiple sites. One patient had chronic typhoid infection. The incidence of clonorchis infestation and primary pyogenic cholangitis in these 100 patients was not different from that of our general hospital population. PMID- 7470818 TI - The PO2 in venous valve pockets: its possible bearing on thrombogenesis. AB - The PO2 in the lumen and valve pockets of veins in 2 patients and 8 dogs was measured during streamline blood flow and in conditions of intermittent pulsatile flow. The blood within the valve pockets became rapidly hypoxic when undisturbed during streamline flow (i.e. when "static"), but the PO2 in the pockets rose to that of the lumenal blood when the column of venous blood was made to pulsate and so empty the valve pockets at short intervals. The observations suggest that the endothelium covering the valve cusps is entirely dependent on pocket or lumenal blood for its oxygen supply. The endothelium facing the pocket can therefore become hypoxic during non-pulsatile blood flow when that facing the lumen is adequately to oxygenated. Early thrombus formation was seen to develop on a valve cusp after only 2 h non-pulsatile flow. The demonstration that localized hypoxaemia occurs readily and can produce endothelial damage, in circumstances and situations where thrombi are commonly found to originate, is additional circumstantial evidence that hypoxia may trigger thrombogenesis. PMID- 7470819 TI - Expanded polytetrafluoroethylene grafts for severe lower limb ischaemia. AB - Three years' experience with the use of expanded polytetrafluoroethylene grafts for severe lower extremity ischaemia are reported; 44 femoropopliteal bypasses were performed, 35 for limb salvage. The prosthesis was used when no saphenous vein was available (23) or to shorten operation time in infirm patients (21). Of 41 patients with a mean age of 70 years, 19 had significant ischaemic heart disease and 10 were diabetic. The mean follow-up period for patient grafts is now 16 months (4--39 months), and the overall life-table cumulative patency is 59 per cent at 6 months, 52 per cent at 1 year and 40 per cent at 2 years. No correlation was demonstrated between patency and run-off, diabetes or smoking, but above-knee grafts may do better than those below the knee. Twenty grafts have occluded leading to 12 amputations, 3 further reconstructions and 1 death. Nine other deaths have occurred from causes not directly related to the lower limb ischaemia, 4 perioperatively and 5 later. The graft described were used in elderly unfit patients, most of whom would have had amputations before the introduction of polytetrafluoroethylene grafts. The results presented support the continued evaluation of this prosthesis in such selected cases. PMID- 7470820 TI - Viscid faecal masses and acute appendicitis. AB - The luminal contents of 200 consecutive appendicectomy specimens were examined prospectively to see if there was any association between viscid faecal matter and acute appendicitis. Faecaliths were found in 13.5 per cent of appendices and were associated with acute inflammation. Viscid faecal masses were present in 28.5 per cent of appendices, but were found more frequently in normal appendices. It is concluded that there is no relationship between acute appendicitis and viscid faecal matter in the lumen. PMID- 7470821 TI - Volvulus of the transverse colon. AB - Volvulus of the transverse colon is rare. Seven cases, 6 acute, are presented. Early diagnosis and prompt surgical management prevented complications. At laparatomy, extended right hemicolectomy or end colostomy and distal mucous fistula are the procedures of choice. Non-resection was associated with recurrence in three cases. PMID- 7470822 TI - A tube system for the management of ruptured oesophagus. AB - In a patient who has ruptured his oesophagus a major objective should be to prevent access of swallowed material or refluxed gastric contents to the area of the rupture. We describe a tube system, introduced into the oesophagus via a cervical oesophagostomy, which prevents swallowed material from getting to the lower oesophagus and allows a tube to the passed into the stomach for continuous aspiration of gastric contents. To discontinue the diversion, a surgical procedure is not required. The successful use of this tube system in 3 patients is described. PMID- 7470823 TI - The influence of an intact pylorus on postprandial enteroglucagon and neurotensin release after upper gastric surgery. AB - Upper partial gastrectomy for carcinoma of the gastric cardia or fundus is unavoidably accompanied by truncal vagotomy, so it is orthodox practice to carry out pyloroplasty with this procedure. In order to assess the effect of leaving an intact pylorus in this operation, 21 patients were investigated with an oral hypertonic load (200 ml 50 per cent glucose solution), and blood was taken at intervals during the test, for the ileal gut hormones, enteroglucagon and neurotensin, as well as for measurements of haematocrit and blood glucose. Clinical features of dumping during the test were sought. There were 7 patients with upper partial gastrectomy, truncal vagotomy and an intact pylorus, 7 patients without a pylorus (total or near total gastrectomy) and 7 patients not subjected to gastric surgery. No significant difference was found in any of the measurements between gastrectomy patients with an intact pylorus and those draining through an enterostomy. However, both these groups differed significantly from normal controls in having greater rates of rise and higher peak values of enteroglucagon, neurotensin, haematocrit and blood glucose, together with a more frequent occurrence of dumping symptoms during the test. As the above measurements probably reflect transit of glucose into the small intestine, the findings in this study suggest that leaving an intact unstenosed pylorus is unlikely to lead to gastric stasis following upper partial gastrectomy. PMID- 7470824 TI - Highly selective vagotomy and pyloric dilatation for duodenal ulcer with stenosis. AB - This paper presents the results obtained in 15 patients with duodenal ulcer and stenosis who were treated more than 3 years ago by highly selective vagotomy (HSV) and dilatation of the stenosis without a gastric drainage procedure. Patients were taking solid food 3--6 days postoperatively and were discharged after 7--14 days. There have been no recurrent stenoses. Fourteen of the 15 patients were Visick grade 1 or 2 at their last visit. One patient has a recurrent ulcer, but no restenosis. Barium meals performed on 6 patients with severe stenosis preoperatively showed satisfactory gastric emptying 1--3 years postoperatively. HSV and pyloric dilatation seems to be a safe and effective procedure for the treatment of pyloric stenosis due to chronic duodenal ulceration. PMID- 7470825 TI - Retroperitoneal fibroses--an association with hyperuricaemia. AB - A review of a small series of cases with idiopathic retroperitoneal fibrosis revealed that two of the patients had been under treatment for gout prior to developing retroperitoneal fibrosis. Investigation showed that other patients in the series had hyperuricaemia. Examination of the retroperitoneal fibrotic tissue demonstrated the presence of urate crystals in unusually high numbers. It is suggested that the pathological process of retroperitoneal fibrosis in these cases may have been initiated by the deposition of urate crystals in the retroperitoneal connective tissue and that the same process may be related to the causation of idiopathic hydrocoele. PMID- 7470826 TI - Gallstone ileus complicating terminal ileal Crohn's disease. AB - Two cases are reported of gallstone ileus complicating terminal ileal Crohn's disease. Both patients had a prolonged history of Crohn's disease without the need for surgical intervention. They were admitted with small bowel obstruction which failed to resolve on conservative management as is usually the case. The features of these two cases that may lead to the development of this complication and the pointers in diagnosis are discussed. PMID- 7470827 TI - Dissolution of common bile duct stones. AB - Retained common duct stones can now be treated by a number of non-operative techniques, including dissolution by cholesterol solvents, simple irrigation, stone extraction via the T tube tract and by endoscopic sphincterotomy. This paper reviews the various solvents available for stone dissolution and its physicochemical basis. When a retained stone is found simple irrigation with saline will be successful in about 50 per cent of patients. If this fails, stone extraction via the T tube tract or by endoscopic sphincterotomy are now the methods of choice, with a 90--95 per cent chance of success. Stone dissolution by cholic acid is slow and is only successful in about 60 per cent of patients but can be of value when stone extraction is unavailable or unsuccessful. PMID- 7470828 TI - Arginine vasopressin release following surgical operations. AB - A study of plasma arginine vasopressin (AVP) concentrations in 16 patients undergoing uncomplicated cholecystectomy was carried out to determine the relative importance of the various stimuli to AVP secretion during and after an operation. Eight of the 16 patients were given a high epidural blockade from T4 and T12 and blood samples were taken before, at frequent intervals during and for two days after operation for determination of plasma AVP and osmolarity; systolic blood pressure was measured each time a blood sample was taken. In the 16 patients there was a 1.8-fold rise of plasma AVP after induction of anaesthesia (from 1.7 pmol/l median basal value to 13.1 pmol/l) and an 8.8-fold rise after skin incision, the highest concentration during operation being 102 pmol/l. There was no significant difference in AVP concentrations, urine volumes or plasma osmolality in the control and epidural groups. Systolic blood pressure fell in all patients during operation by between 14 and 54 per cent, the fall being significantly greater in the epidural group. Notwithstanding the evidence that certain nervous impulses do cause an increased secretion of AVP during operation, these results clearly show that in the absence of such stimuli there is still an increased secretion, almost certainly caused by a fall in the blood pressure. PMID- 7470829 TI - Excision of bullae without pleurectomy in patients with spontaneous pneumothorax. AB - In a retrospective review of 294 patients with spontaneous pneumothorax (398 episodes), 45 patients came to surgical treatment. The period of the study was 7 1/2 years, with a mean follow-up of 3 years in which there was no recurrence of pneumothorax after surgery. Operation consisted of simple excision of bullous lesions, usually located at the apex of the upper lobe, without pleurectomy. This simplified surgical approach has met with excellent result, indicating that pleurectomy is unnecessary in the treatment of this disease. PMID- 7470830 TI - Sequential changes in susceptibility to Treponema pallidum of rabbits previously infected with Treponema paraluis-cuniculi. AB - Rabbits immunised with virulent Treponema paraluis-cuniculi were challenged intradermally with graded doses of Treponema pallidum at three, five, seven, 12, and 30 months to ascertain the level of protection to T pallidum at various intervals after immunisation.Rabbits challenged at three months after immunisation showed no protection against T pallidum and developed syphilitic lesions significantly faster than the control rabbits, which suggests that the former rabbits were immunosuppressed. Some protection was evident at five and seven months after immunisation, as fewer inoculation sites developed syphilitic lesions with challenges of 10(3), 10(2), and 10 T pallidum and lesions developed significantly slower with 10(6) challenge. Two rabbits showed significant protection at 12 months after immunisation but a third, presumably still immunosuppressed, developed lesions significantly faster than the control rabbits after challenge. At 30 months after immunisation one rabbit was completely protected and developed no lesions after challenge; the other rabbit showed only partial protection against challenge with 10(4), 10(3), and 10(2) but complete protection against challenge with 10 T pallidum.T paraluis-cuniculi appeared to induce a state of immunosuppression by three months after infection; in one rabbit this may have been 12 months. In most immunised rabbits, however, limited cross-protection to low challenge doses of T pallidum developed by five months and was also detectable at seven and 12 months. Only one rabbit was completely resistant to challenge with 10(4)T pallidum after 30 months and another was only partly immune. Thus, T paraluis-cuniculi infection does not produce a rapid pronounced cross-immunity to T pallidum in rabbits, which may thus limit its usefulness as a vaccine against syphilis. PMID- 7470831 TI - Immunoglobulins in the cerebrospinal fluid of patients with late syphilis. AB - The concentrations of immunoglobulins G, A, and M were measured in 62 samples of cerebrospinal fluid from 35 patients with asymptomatic neurosyphilis, 18 with symptomatic neurosyphilis, and nine with late syphilis of other organs. The most frequent and highest increase in IgG and IgA occurred in patients with symptomatic neurosyphilis. IgM was found in only five patients, but its presence, together with an increase in IgG and IgA concentrations, appears to indicate activity of the pathological process and a poor prognosis for the course of the disease. PMID- 7470832 TI - An atypical case of secondary syphilis. AB - A 27-year-old man with secondary syphilis had unusual cutaneous lesions of the ear and scalp. PMID- 7470833 TI - Microbiological, serological, and histopathological features of experimental Chlamydia trachomatis urethritis in chimpanzees. AB - A laboratory-passaged genital strain of Chlamydia trachomatis and two unpassaged genital strains from patients with nongonococcal urethritis were inoculated intraurethrally into three young male chimpanzees. Chlamydia were recovered from the urethra of two animals and specific antibody responses were detected in all of them. Furthermore, a urethral polymorphonuclear leucocyte response, but not an overt discharge, occurred in all the chimpanzees about 1-2 weeks after inoculation. None of these events occurred in a chimpanzee inoculated with medium only. At necropsy three months after inoculation the submucosa of the urethra of one chimpanzee was densely infiltrated with small round cells. This suggests that a similar chronic lymphocytic response may occur in human chlamydial infection of the urethra. PMID- 7470834 TI - Chlamydia trachomatis as a cause of acute perihepatitis associated with pelvic inflammatory disease. AB - Of four cases of acute salpingitis and perihepatitis confirmed by laparoscopy Chlamydia trachomatis was cultured from the cervix only in two and from both the cervix uteri and the Fallopian tubes in two; the latter finding has not been reported from cases with combined salpingitis and perihepatitis (Fitz-Hugh-Curtis syndrome). Since gonococci, other aerobic, or anaerobic bacteria were not isolated from the Fallopian tubes, an aetiological relationship between C trachomatis and the Fitz-Hugh-Curtis syndrome is suggested. PMID- 7470835 TI - Chlamydial infection in homosexual men. Frequency of isolation of Chlamydia trachomatis from the urethra, ano-rectum, and pharynx. AB - Urethral, rectal, and pharyngeal material from 150 men who had had exclusively homosexual contact and who consecutively attended a sexually transmitted diseases clinic was cultured for Chlamydia trachomatis. The organism was isolated from at least one site in 15 (105) patients. The isolation rates from the urethra, rectum, and pharynx were 6 . 7%, 4%, and 1 . 3% respectively. PMID- 7470836 TI - Isolation of adenovirus type 19 from the male and female genital tracts. AB - During routine screening for genital herpes simplex virus infection in patients attending a sexually transmitted diseases clinic adenovirus type 19 was isolated from both men and women. Peak incidences of genital infection with adenovirus type 19 corresponded with those of eye infection with the same virus in the general community. Thus, the relationship between genital and eye infection with adenovirus, the part played by genital infection in its dissemination, and the clinical symptoms it may produce need further study. PMID- 7470837 TI - Experimental infection of man with rabbit-virulent Treponema paraluis-cuniculi. AB - Virulent Treponema paraluis-cuniculi was inoculated intradermally into the arm of a human volunteer and into the shaved backs of 10 rabbits. An identical, but heat killed, preparation was inoculated into the opposite arm of the volunteer as control. A superficial and transient infection developed in the volunteer, shown by a small zone of erythema that persisted for 24 days. The control preparation caused a smaller zone of erythema that disappeared after five days. A very poor immune response was detected by standard serological tests for syphilis. The inoculated rabbits developed lesions about six days after infection and seroconverted by 84 days. The poor antitreponemal antibody response to T paraluis cuniculi infection in the volunteer suggests that this naturally attenuated treponeme may not be suitable as a vaccine against infection with t pallidum in humans. PMID- 7470838 TI - Cerebral disorder in alcoholism: syndromes of impairment. PMID- 7470839 TI - The epidemiology of aphasic and cognitive impairment in stroke: age, sex, aphasia type and laterality differences. AB - Sex differences and age distribution are examined in various types of aphasia and in right hemisphere-damaged patients due to acute infarct. The language and performance deficits were measured by a standardized test allowing us to group patients meaningfully. The higher male to female ratio of aphasia is shown to relate to a similar sex distribution in infarcts rather than to sex differences in cerebral organization. Broca's aphasics were somewhat younger than the other groups and the slight difference may be related to pathophysiological factors favouring embolic strokes in the anterior territory of the middle cerebral artery. Sex and aphasia types were not significantly different in various age groups except about 50 years of age where the female aphasics reversed the ratio. PMID- 7470840 TI - Cerebral potentials evoked by muscle stretch in man. AB - The cerebral potentials evoked by passive plantar flexion movements of the ankle were recorded by scalp electrodes in normal human subjects. The potential consisted of a biphasic positive wave (P45, P65), a prominent negative wave (N90), and a subsequent positive-negative-positive complex (P120, N145, P190). The components of the potential were of largest amplitude at the vertex, small displacements of the active recording electrode resulting in a marked decrease in amplitudes. The amplitudes of the components were also profoundly affected by changes in the repetition rate of the stimulus and by changes in the acceleration of the stretching movements; they were little affected by changes in the extent or peak velocity of stretch provided that acceleration remained constant. Potentials of similar morphology but of slightly shorter latency could be evoked by percussion of the tendon of tibialis anterior and by electrical simuli delivered through a microelectrode inserted into a pure muscle nerve fascicle of the peroneal nerve innervating one of the pretibial flexor muscles. The results of nerve block experiments (ischaemic/pressure block above the ankle; pressure block and local anaesthetic block of the peroneal nerve at the fibular head) suggest that the afferents responsible for evoking the cerebral potential arose from muscle mechanoreceptors. It is concluded that information about muscle stretch arising from muscle mechanoreceptors (probably muscle spindle endings) reaches the cerebral hemispheres of man at short latency and that the resulting potentials can be readily detected using scalp electrodes. PMID- 7470841 TI - Reaction time in Parkinson's disease. AB - Both reaction time and movement time tend to be prolonged in Parkinson's disease, but they are often impaired independently of each other. Prolongation of RT is relatively slight, while MT undergoes more substantial and consistent disturbance. Choice RT and kinaesthetic RT do not have any advantage over simple visual RT as measurements of neurological deficit in parkinsonism, since they are all impaired to the same extent. MT is more useful than RT as an objective indicator of therapeutic efficacy, but further studies of RT (with tests requiring programming of displacement, velocity, and accuracy) may provide insights into the nature of the central motor disorder in Parkinson's disease. PMID- 7470842 TI - Autonomic neuropathy in experimental allergic neuritis: an electrophysiological and histological study. AB - Electrophysiological and histological studies have been performed on the vagus and splanchnic nerves of guinea pigs with experimental allergic neuritis. Slowing of conduction and dispersion of the compound action potential were consistent with the pathological changes of demyelination. In teased single fibre preparations, axonal degeneration was found more frequently than was expected and in the splanchnic nerves, the unmyelinated fibres appeared to have been indirectly involved. These findings in experimental allergic neuritis are relevant to the pathogenesis of autonomic dysfunction in the Landry-Guillain Barre syndrome. PMID- 7470843 TI - Lexical or orthographic agraphia. AB - A case of agraphia was studied in the framework of the information-processing approach and in relation to comparable studies on alexia. Three dissociations are of interest in this patient's picture: (a) in language production: impairment of writing without impairment of speech; (b) in writing: impairment of spelling without impairment of phoneme-grapheme conversion; (c) in visible language: impairment of writing without equivalent impairment of reading. The implications of these dissociations for any model of linguistic processes are discussed. PMID- 7470845 TI - Familial dementia of adult onset with pathological findings of a 'non-specific' nature. AB - A family is described in which 4 of 10 siblings developed a dementing illness that culminated in death within five to six years of onset. The pathological findings in 3 members were strikingly similar, and consisted of widespread nerve cell loss and astrocytosis within the cerebral cortex, status spongiosus within the outer cortical layers and, in 2, nerve cell loss and astrocytosis within the dorsomedial nucleus of the thalamus. It is concluded that the disorder described in this report does not conform precisely to any of the currently recognized categories of familial dementing disease. PMID- 7470844 TI - Arnold-Chiari malformation: improvement in eye movements after surgical treatment. AB - We studied abnormalities of eye movement in five patients with Arnold-Chiari malformation before and after suboccipital decompression. Before operation all patients complained of oscillopsia at rest and visual blurring of fixated targets during and immediately after their heads were in motion. Three patients had downbeat nystagmus, one patient had horizontal nystagmus in the primary position and on eccentric gaze and one patient had rotatory nystagmus on lateral gaze only. Smooth pursuit, optokinetic nystagmus and fixation suppression of vestibular nystagmus were markedly impaired. Within two months after operation nystagmus in the primary position had resolved in three patients, and four had complete to near-complete resolution of nystagmus in all gaze positions over subsequent months. Oscillopsia decreased concurrently. affording significant functional visual improvement. One patient had only slight functional improvement and continued to have prominent nystagmus seven years after operation. Smooth pursuit, optokinetic nystagmus and fixation suppression of vestibular nystagmus improved in all patients, but none had complete recovery at the higher velocities of stimulation. They still complained of visual blurring associated with head motion, probably due to the poor fixation suppression of vestibular nystagmus. Our findings suggest that abnormalities of eye movement in Arnold-Chiari malformation are due to compression of the herniating cerebellum against the caudal brain-stem and not a congenital aberrancy of oculomotor pathways. Nystagmus resolved more readily than pursuit or fixation suppression abnormalities, but most patients showed gradual improvement in all abnormalities four months after operation. PMID- 7470846 TI - Distribution of cerebral lesions in acquired hepatocerebral degeneration. AB - The brain lesions of our patients with chronic portal-systemic venous shunts were much the same regardless of the presence or absence of primary liver disease. Alzheimer's type 2 abnormality of astrocytes and demarcated areas of spongy degeneration were found, mainly in the grey matter, with more severe involvement of the basal ganglia and the deeper layers of the frontal, parietal and occipital cortex. The spongy degeneration showed an especially close correlation with the arterial blood supply, being greatest in borderland areas that fall between the regions usually supplied by one or other of the major cerebral arteries. Previous investigations have not been primarily concerned with selective vulnerability so that comparisons are difficult, but on review, there is evidence that this pattern of involvement is a feature of both Wilson's disease and acquired hepatocerebral degeneration. The toxic effect of the portal blood is greatest in the grey matter, probably because of the greater metabolic activity there, and the distribution of lesions within this area of greater vulnerability appears to be further influenced by circulatory factors. PMID- 7470847 TI - Delayed and enhanced long latency reflexes as the possible cause of postural tremor in late cerebellar atrophy. AB - The pathophysiology of postural tremor was studied in 7 patients with cortical cerebellar atrophy, and compared with the responses of 14 healthy control subjects to the same tests. Both tibial nerves were simultaneously and selectively stimulated in the fossa poplitea. EMG was recorded from agonist gastrocnemius muscles and from the antagonistic anterior tibial muscles. Displacement of the centre of foot pressure, inclination of trunk and head in the anteroposterior direction, and the ankle angle were also measured. Patients and controls both exhibit a synchronized discharge in the anterior tibial muscle (antagonist) with a latency of 120 ms to stimulus onset (tib1). Tib1 is shown to be a segmental stretch reflex elicited by the contraction of the gastrocnemius (agonist). A later, presumed long-loop response occurs after another 120 ms both in gastrocnemii and anterior tibial muscles in the normal subjects. This latency, and the amplitude of the late reflex, are increased in the patient group. The synchronization of delayed long-loop reflexes and a stretch response of the gastrocnemius in response to tib1 terminate the first cycle of the postural tremor which thereafter continues by way of the same mechanism generating a contraction of the anterior tibial muscle. Postural tremor can thus be synchronized by a single bilateral electrical stimulus and can even be elicited in incipient cases of the disease. With further progression of the cerebellar atrophy the dominant frequency of the postural tremor decreases along with an increase of long-loop latencies. PMID- 7470848 TI - Neurolinguistics: theoretical and evolutionary perspectives. PMID- 7470849 TI - The use of averaged electroencephalic response techniques in the study of auditory processing related to speech and language. PMID- 7470850 TI - Hemispheric specialization for temporal information: implications for the perception of voicing cues during speech perception. PMID- 7470851 TI - Event-related potentials in neuropsychological studies of language. PMID- 7470852 TI - Behavioral and neural analyses of connotative meaning: word classes and rating scales. PMID- 7470853 TI - Linguistic meaning related differences i evoked potential topography: english, swiss-german, and imagined. PMID- 7470854 TI - Reading between the lines: event-related brain potentials during natural sentence processing. PMID- 7470855 TI - Electroencephalographic and dichotic indices of cerebral laterality in stutterers. PMID- 7470856 TI - Descending projections of the locus coeruleus and subcoeruleus/medial parabrachial nuclei in monkey: axonal transport studies and dopamine-beta hydroxylase immunocytochemistry. AB - Spinal projections originating in the dorsolateral pons in the ventral locus coeruleus and in the subcoeruleus and medial parabrachial nuclei were identified in monkeys (Macaca fascicularis) using the retrograde horseradish peroxidase tracing technique. Anterograde autoradiographic tracing studies were then carried out to determine the brain stem and spinal cord terminations of the neurons. Finally, results obtained with the axonal transport tracing methods were compared with the pattern of staining for noradrenergic cells and terminals revealed immunocytochemically with an antibody to dopamine-beta-hydroxylase (DbetaH), the synthesizing enzyme for norepinephrine. The major findings of these studies are that two presumed noradrenergic cell groups of the dorsolateral pons, one corresponding to the nucleus locus coeruleus, the second to the subcoeruleus/medial parabrachial nuclei, give rise to descending projections. They differ significantly in their patterns of termination in the lower brain stem and spinal cord. Among the major terminations of the locus coeruleus pathway are projections to parasympathetic neurons of the dorsal motor nucleus of the vagus, the region of the nucleus ambiguus, and the sacral spinal cord. The terminations of the descending subcoeruleus/medial parabrachial pathway, in contrast, include projections to sympathetic preganglionic neurons of the intermediolateral cell column of the thoracic cord and heavier projections to somatic cranial nerve nuclei. Both pathways have additional widespread and bilateral terminations in various nuclei of the reticular formation, in the spinal dorsal horn (including the marginal zone), in the region around the central canal and in the ventral spinal gray matter. Since the origins and terminations of both these pathways correspond closely to the locations and patterns of terminations of noradrenaline-containing neurons, demonstrated here with DbetaH immunocytochemistry, norepinephrine (or epinephrine) is suggested to be the transmitter in both these descending systems. PMID- 7470857 TI - Cholinergic projections from magnocellular nuclei of the basal forebrain to cortical areas in rats. AB - Acetylcholinesterase (AChE) and choline acetyltransferase (CHAc) activities were studied by quantitative histochemical (AChE) as well as biochemical methods (AChE, ChAc) in certain cortical brain areas in rats after stereotaxic lesions had been placed in several structures of the basal forebrain. After lesioning the magnocellular nuclei of the substantia innominata (nuc. basalis Meynert, NBM) the activities of AChE and ChAc decreased to moderate or low residual values in the ipsilateral cortical areas. This indicated that cholinergic pathways were directly linked to frontal, sensory-motor, auditory and visual cortex. After lesions of the globus pallidus the decrease in cortical AChE activity was less pronounced. Lesions of the caudate, accumbens or entopeduncular nucleus did not influence the cortical AChE activities. The results are discussed with respect to the similarity of the organization of the cholinergic projection to the cortex arising from NBM cells and the monoaminergic system which innervates the cortex. It is suggested that both neurotransmitter systems by their interaction might modulate and control cortical information processing and behavior in a manner analogous to the control of peripheral activity by the sympathetic and parasympathetic system. PMID- 7470858 TI - An allometric study of the area dentata in the rat and mouse. AB - The volumes of the hilus, stratum granulosum, stratum moleculare and the zones of the stratum moleculare corresponding to the terminal fields of the perforant paths and the ipsilateral-commissural projections have been estimated in the area dentata of Wistar rats and DBA/2J mice, stained according to the Timm technique. The density of granule cells in the stratum granulosum and synaptic contacts in the stratum moleculare have been estimated from low power electron micrographs using stereological techniques. The volume and density parameters have been used to calculate the total number of granule cells and synaptic contacts in the fasciae dentatae of these two species. It is concluded that the number of synaptic contacts is proportional to the volume of the fascia dentata while the number of granule cells is proportional to the surface area of the fascia dentata and that the fasciae dentatae in these two species are isometric forms. The implications of these relationships are discussed with regard to the effect of the size of the fascia dentata on information processing in this structure. A significant difference existed between the ratio of the volumes of the hila and the ratio of the volumes of the fasciae dentatae in the two species studied. However, the volume of the deep zone of the stratum moleculare fasciae dentatae, the terminal zone of the hilar afferents was proportional to the volume of the hilus. The same density of synaptic contacts in the deep zone of the strata molecularia of both species, therefore, indicates a proportionality between the volume of the hilus and the number of synaptic contacts made by the hilar afferents. These observations are discussed with respect to the manner by which the maintenance of the observed proportionalities and the relative differences in the size and number of neurons in the subregions of the area dentata may be involved in the modification of the form and function of this region during phylogenesis. PMID- 7470859 TI - The contribution of brain stem catecholamine cell groups to the innervation of the sympathetic lateral cell column. AB - Data are presented on the quantitative distribution of individual catecholamines, noradrenaline (NA), dopamine (DA) and adrenaline (A) determined within specific microdissected areas of the third thoracic segment of the cat spinal cord, using a radioenzymatic assay. NA was found in much higher concentrations than the other catecholamines with highest levels in medial-ventral grey matter (754 +/- 56 ng/100 mg protein) and in dorsal and intermediolateral grey matter. DA was present in much smaller amounts, but was relatively concentrated in medial and dorsal grey matter (up to 76 +/- 1 ng/100 mg protein). Very little A was detected ( less than 17 ng/100 mg protein). Experiments were carried out to determine the contribution made by various brain stem cell groups to the catecholamine innervation of the spinal cord, and in particular the sympathetic lateral column. The concentrations of the catecholamines was determined in 5 groups of animals, 3 5 days after decerebration, or decerebration together with bilateral lesions of the A1, A2, A5 or A6 groups. A major and selective NA input to the sympathetic lateral column from the A1 cell group was revealed. There may also be a small contribution from the A2 and A6 groups, but none was shown from the A5 group. The A6 group was also shown to provide a selective noradrenergic input to the ventral horn. Some evidence is presented for the origin of noradrenergic inputs to other regions, e.g. dorsal horn. The possibility of a separate dopaminergic innervation to specific regions of the spinal cord is discussed. PMID- 7470860 TI - Ultrastructural analysis of medial brain stem afferents to the superficial dorsal horn. AB - Axonal projections of medial brain stem areas rich in serotonin-containing neurons were identified in layers I and II of cat medullary dorsal horn using EM autoradiography. Following [3H]amino acid injections into the brain stem, labeled axonal endings were found throughout layers I and II but were most numerous in layer I. Three different morphological types of endings could be distinguished. Each type resembled serotonergic axonal endings identified in previous experiments. The labeled endings formed both symmetrical and asymmetrical synapses on dendritic shafts and spines and occasionally on a neuronal soma suggesting that the major site of action of the descending serotonergic afferents is on the neurons in layers I and II. PMID- 7470861 TI - Alpha-bungarotoxin binding in cat carotid body. PMID- 7470862 TI - Model of behavioral modulation of the hippocampal CA1 region of the rat. PMID- 7470863 TI - Galactose oxidase labeling of membrane proteins from human brain white matter. AB - The use of galactose oxidase (EC 1.1.3.9) and tritiated sodium borohydride for labeling of membrane glycoproteins, described by Gahmberg and Hakomori, has previously been applied to the study of myelin glycoproteins of experimental animals. Rat brain myelin glycoproteins have been studied by sequential lectin affinity of chromatography and recently the lectin-binding capacity of rat central nervous system myelin glycoproteins has been characterized. Complex heterogeneity of the glycoprotein pattern of rat central nervous system myelin has been reported, and so a variety of glycoproteins can be expected to exist in human white matter membranes. Application of the galactose oxidase procedure to the study of human brain membranes could be useful in research concerning certain neurological diseases if the properties of autopsy brain material are taken into account. In this study, membrane proteins of human autopsy brain white matter were subjected to the galactose oxidase/NaB3H4 labeling procedure and the membrane labeled by this method or by the [3H]acetic anhydride techniques were studied by lectin affinity chromatography using Lens culinaris phytohemagglutinin (lentil lectin) attached to Sepharose 4B beads. PMID- 7470865 TI - Effects of restricted lesions of the chick forebrain on the acquisition of filial preferences during imprinting. AB - The effects of placing bilateral lesions in that part of the chick brain (IMHV) which was previously been implicated in imprinting, was studied in young domestic chicks. Twenty-four dark-reared chicks were matched in pairs on the basis of their approach activity during a 30 min period of exposure to one of two visual imprinting stimuli. Both members of the chick pair were then anaesthetized and bilateral lesions were made by radio-frequency coagulation in the IMHV of one chick; the other chick served as a sham-operated control. On the following day each chick was exposed for 2.5 h to the imprinting stimulus to which it had previously been exposed. After training, the preferences of all chicks were measured by comparing their approach to the training stimulus with that to the second stimulus. Sham-operated chicks showed a strong preference for the training stimulus; lesioned chicks showed none. Subsequently the latency of each chick to approach and accurately peck a shiny rod was measured. The two groups of chicks did not differ significantly in this test of visuomotor coordination. The area of tissue damaged by the lesion was reconstructed: IMHV was severely damaged with relatively little damage to other areas of the brain. PMID- 7470864 TI - Aspects of salt/water balance after cerebroventricular infusion of angiotensin II. PMID- 7470866 TI - Deficits in contralateral and ipsilateral forepaw motor control following unilateral motor cortical ablations in rats. AB - Two force transducers which were interfaced with a minicomputer were used as operant manipulanda to assess changes in peak force and rate of response produced by unilateral motor cortical lesions in rats. Spatial arrangement of the manipulanda permitted a subject to respond on Transducer I exclusively with his left forepaw and on Transducer II exclusively with his right. Permanent deficits as measured by both force and rate of response were observed for the limb contralateral to the lesion. In the forelimb ipsilateral to the lesion rate of response was severely depressed immediately following the lesion and, although there was marked recovery during the 10-12 days following surgery, post-lesion rates never attained pre-lesion levels. PMID- 7470867 TI - Estimation of numbers of vomeronasal synapses in the glomerular layer of the accessory olfactory bulb of the mouse at different ages. PMID- 7470868 TI - Prevention of renal hypertension and of the central pressor effect of angiotensin by ventromedial hypothalamic ablation. AB - Various lines of research have implicated the central nervous system in the development of renal hypertension. The ablation of a periventricular region surrounding the anteroventral third ventricle (AV3V) has been shown to block the development of renal hypertension. Because of hemodynamic effects produced by AV3V electrical stimulation can be abolished by a midline lesion of the ventromedial hypothalamic--median eminence region (VMH-ME), the effect of VMH-ME ablation on the development of renal hypertension was studied. Following recovery from surgery that destroyed the VMH-ME region the lesioned rats and controls were subjected to unilateral nephrectomy and figure-of-eight wrapping of the remaining kidney. Control animals developed renal hypertension but those with VMH-ME lesions did not. When the pressor response produced by intracerebroventricular injections of angiotensin II was studied, it was found that rats with VMH-ME lesions, as compared to neurologically intact animals, showed significantly attenuated responses. The data suggest that a neural system related to cardiovascular control descends through the VMH-ME region and that the integrity of this pathway is necessary for the development of renal hypertension. PMID- 7470869 TI - Apomorphine-haloperidol interactions: different types of antagonism in cortical and subcortical brain regions. AB - Apomorphine, a dopamine (DA) receptor stimulant induced a dose-dependent decrease in the content of the dopamine metabolites 3,4-dihydroxyphenylacetic acid (DOPAC) and homovanillic acid (HVA) in the olfactory tubercle and striatum of rats, but it was ineffective in the frontal cortex. Apomorphine also antagonized the haloperidol-induced accumulation of DOPAC and HAV in the olfactory tubercle and striatum, whereas in the frontal cortex it antagonized the effects of haloperidol on DOPAC but not on HVA. The total (free and conjugated) content of DA metabolites was measured in these experiments. PMID- 7470870 TI - Eye and head movements during vestibular stimulation in the alert rabbit. AB - Rabbits passively oscillated in the horizontal plane with a free hand tended to stabilize their head in space (re: earth-fixed surroundings) by moving the head on the trunk (neck angular deviation, NAD) opposite the passively imposed body rotation. The gain (NAD/body rotation) of head stabilization varied from 0.0 to 0.95 (nearly perfect stability) and was most commonly above 0.5. Horizontal eye movement (HEM) was inversely proportional to head-in-space stability, i.e. the gaze (sum of HEM, NAD, and body rotation) was stable in space (regardless of the gain of head stabilization). When the head was fixed to the rotating platform, attempted head movements (head torque) mimicked eye movements in both the slow and fast phases of vestibular nystagmus; tonic eye position was also accompanied by conjugate shifts in tonic head torque. Thus, while eye and head movements may at times be linked, that the slow eye and head movements vary inversely during vestibular stimulation with a free head indicates that the linkage is not rigid. Absence of a textured stationary visual field consistently produced a response termed 'visual inattentiveness,' which was characterized by, among other things, a reduction of head and gaze stability in space. This behavioral response could also be reproduced in a subject allowed vision during prolonged vestibular stimulation in the absence of other environmental stimuli. It is suggested that rabbits optimize gaze stability (re: stationary surroundings), with the head contributing variably, as long as the animal is attending to its surroundings. PMID- 7470871 TI - Projections of caudal fasciculus gracilis to nucleus gracilis and other medullary structures, and Clarke's nucleus in the rat. AB - The extent of the projection of fasciculus gracilis from low thoracic (T12--13) spinal cord, to nucleus gracilis (NG) was studied by the Nauta--Gygax method in the rat in order to: (1) help define projection fields of fasciculus gracilis hindlimb and tail afferents along the longitudinal extent of NG; and (2) broaden the comparative data on the projection of the lower body to NG via fasciculus gracilis, already known in cat and monkey. Results showed that: (1) degenerated fasciculus gracilis axons are situated mainly dorsal to the corticospinal fibers, in the dorsal funiculus; (2) terminal projection fields were located at all levels of ipsilateral NG--caudal to the obex, terminal debris was confined to the medial two-thirds of NG--rostrad of the obex, in oral NG, degeneration was of finer caliber and projection fields more diffuse; and (3) ipsilateral degeneration was also seen at the level of a portion of Clarke's nucleus, and, in commissural and lateral cuneate nuclei and medial reticular formation in medulla. PMID- 7470872 TI - Corticopontine projections from the cingulate cortex in the rhesus monkey. AB - The corticopontine projections of the cingulate cortices were investigated in the rhesus monkey with the use of autoradiography. A well-organized topography of projections was observed with anterior cingulate cortex projecting to the medial part of the pontine gray matter and posterior cingulate cortex projecting to the lateral part. Together these projections form a circle of termination around the periphery of the pontine gray matter. PMID- 7470873 TI - A new microelectrophoretic procedure for delivery of horseradish peroxidase. AB - A new procedure for microelectrophoretic delivery of horseradish peroxidase (HRP) is described. A dilute solution (1.5--3% w/v) of fluorescein isothiocyanate conjugated HRP is deposited according to a standard microelectrophoretic method and the tissue reacted according to a sensitive histochemical procedure. A major advantage of this method is the reduced secondary diffusion of the enzyme present at the injection site. PMID- 7470874 TI - Responses in the rabbit medial preoptic region to stimulation of the fimbria. PMID- 7470875 TI - Polarographic assay of iontophoretically applied dopamine and low-noise unit recording using a multibarrel carbon fibre microelectrode. PMID- 7470876 TI - Influences of cerebellar hemispherectomy upon cortical potentials preceding visually initiated hand movements in the monkey. AB - With chronically implanted electrodes, surface and depth potentials of the premotor and motor cortices were recorded on hand movements in response to a visual stimulus in monkeys, and influences of cerebellar hemispherectomy were examined upon visually initiated premovement cortical potentials. Early, surface positive--depth negative premovement potentials emerged in the cortices on both sides, and following surface negative--depth positive premovement potentials appeared in the motor cortex contralateral to the moving hand. Cerebellar hemispherectomy contralateral to the motor cortex eliminated the following potentials. This suggests the participation of the neocerebellum in preparing the motor cortex for visually initiated movements. PMID- 7470877 TI - Increase in histamine concentrations in discrete hypothalamic nuclei of spontaneously hypertensive rats. PMID- 7470878 TI - Naloxone effects on the blood pressure response induced by thin-fiber muscular afferents. AB - Naloxone effects on the blood pressure level and on the blood pressure responses induced by thin-fiber muscular afferent stimulation were studied in anesthetized, bilaterally vagotomized and carotid sinus nerve-denervated dogs under artificial ventilation. Repetitive pulses of 8 Hz with various intensities were applied to the gastrocnemius nerve for 1 min while monitoring the compound action potentials. The mean arterial pressure significantly (P less than 0.001) rose by 10.95 +/- 1.78 mmHg (mean +/- S.E.) about 5 min after a naloxone injection. Compared with the reflexive response in the control period, the depressor effect significantly decreased by 3.80 +/- 1.06 mmHg, and the pressor effect significantly increased by 3.63 +/- 0.73 mmHg for 30 min after the injection of naloxone. No correlation was found between naloxone effects on the blood pressure level and on the reflex response, indicating an involvement of different mechanisms with these naloxone effects. We suggest that endogenous opiates might participate in the regulation of the blood pressure level, as well as of the blood pressure responses caused by thin-fiber muscular afferents. PMID- 7470879 TI - Developmental changes of synapses in the cerebellar cortex of the rat. A quantitative analysis. AB - Synaptogenesis in the molecular layer of the vermis cortex in Wistar rats between 1 and 25 days after birth was investigated. After staining with OSO4, the following parameters were measured: the density of synaptic profiles; the percentage of the neuropil area occupied by synapses; the mean diameter of the boutons; and the numerical density of synapses in a defined volume. The detailed topographical analysis enabled us to show the following: the first synapses appear diffusely in the molecular layer; after the 10th day a synaptic gradient is present between the depth and the surface area (this gradient can be found by the count of the synapses and by studying the spatial distribution of the synaptic area); this gradient is no longer seen at the 25th day, when the density of synapses is relatively uniform throughout the whole molecular layer. PMID- 7470880 TI - Polyornithine-attached neurite-promoting factors (PNPFs). Culture sources and responsive neurons. AB - We have recently reported the existence within chick embryo heart cell conditioned medium (HCM) of two distinct and independently assayable factors. One agent, ciliary neuronotrophic factor (CNTF), supports the in vitro survival of 8 day chick embryo ciliary ganglionic (CG) neurons. The other factor, polyornithine attachable neurite promoting factor (PNPF) is required for extensive neuritic growth from these same CNTF-supported CG neurons. In the present study we have examined the occurrence of PNPF activity within nearly 100 different conditioned media using our previously described chick CG bioassay system. From this screening we conclude that: (1) PNPF production is a rather widespread property of cultured neural as well as non-neural cells; and (2) the chick bioassay is sensitive to PNPF activity from all the species examined, including mouse, rat, human and chick cells. We next examined the effects of 3 representative PNPF containing conditioned media (from chick heart, mouse Schwann and rat Schwannoma) on neurite production from 3 other peripheral ganglionic neuronal cultures (8-day chick dorsal root, 11-day chick sympathetic, and neonatal mouse dorsal root ganglia) as well as 4 central neuronal cultures (8-day chick embryo telencephalon, optic lobe and spinal cord and neonatal mouse cerebellum). The results of these studies indicate: (1) that the peripheral neurons exhibit a dramatic increase in neurite production in response to PNPF which can be easily recognized both qualitatively and quantitatively; whereas (2) the CNS neurons showed essentially no PNPF-induced increase in neurite production. The sole exception to the latter was the appearance within the chick spinal cord cultures of a neuronal population which extended very long neurites in response to PNPF. PMID- 7470881 TI - The accessory optic system in pigeons: receptive field properties of identified neurons. AB - Visual receptive fields of neurons in the nucleus of the basal optic root were investigated in pigeons. Their projections could be traced to vestibulo cerebellum and oculomotor complex by means of antidromic activation. Units in that nucleus showed large peripheral receptive fields and appeared highly sensitive to moving targets, with the majority displaying axis specificity and direction selectivity. The results seem consistent with the proposed role of the accessory optic system in oculomotor reflexes. PMID- 7470882 TI - The effects of unilateral carotid occlusion on the responses to decapitation in the gerbil brain. AB - The effects of partial ischemia (unilateral carotid artery occlusion) on the metabolic and electrical responses to decapitation were studied in the awake and anesthetized gerbils' brains. The gerbil was connected to the 2 channel fluorometer/reflectometer by flexible light guides implanted above the two hemispheres of the brain. The results show that: (1) the metabolic rate as measured by this present technique is lower in the gerbil than in the rat; (2) the metabolic rate of the ischemic hemisphere is lower as compared to that of the normoxic contralateral hemisphere. PMID- 7470883 TI - The postnatal development of synapses in the visual cortex of the cat and the effects of eyelid closure. PMID- 7470884 TI - Normal cortical responses in ocularly hypopigmented cats. PMID- 7470885 TI - Aromatic alkylamine N-acetyltransferase in regional tissues in developing rat brain. AB - The specific activities of N-acetyltransferase (EC 2.3.1.5) were examined in several regions of rat brain of both sexes at various times after birth. The enzyme activity increased with development in whole brain, hippocampus, midbrain, cerebellum and the remainder of brain, peaking around 36 days of age. Lineweaver Burk plots indicated linear kinetics for N-acetyltransferase in dialyzed supernatant and ammonium sulfate precipitates from the newborn rat brain, whereas enzyme preparation further purified by Bio Gel yielded biphasic kinetics. These data remain consistent with the possibility that there are two forms of N acetyltransferase in rat brain even from birth. PMID- 7470886 TI - Hypomyelinated mutant mice. III. Increased myelination in mutant cerebellum co cultured with normal optic nerve. PMID- 7470887 TI - Evidence for the neuronal origin of brain capillary acetylcholinesterase activity. AB - Acute administration of monosodium-L-glutamate (MSG) to rats induces the degeneration of acetylcholinesterase (AChE)-positive neurons of the area postrema (AP). Chronic treatment with MSG results in the disappearance of AChE activity of area subpostrema (ASP) capillaries. It is concluded that processes of AChE positive AP neurons terminate within the ASP and may contribute to the AChE activity of ASP capillaries. PMID- 7470888 TI - Evidence for a tonic GABAergic control of serotonin neurons in the median raphe nucleus. AB - We examined whether serotonin (5-HT)-containing neurons of the midbrain raphe nuclei are subject to an inhibitory control by GABA. We found that injection into the median raphe nucleus of the GABA antagonists picrotoxin and bicuculline and the GABA agonist muscimol increase and decrease, respectively, the 5-HT turnover and the steady-state content of 5-hydroxyindoleacetic acid. The results provide evidence of a tonic inhibition by GABA of 5-HT neuronal activity in the median raphe nucleus; this inhibitory effect is potentiated by benzodiazepines. PMID- 7470889 TI - An arginine vasotocin-like neuropeptide is present in the nervous system of the marine mollusc Aplysia californica. AB - Radioimmunoassays and high pressure liquid chromatography have been used to demonstrate the presence of an arginine-vasotocin-like peptide (AVT) in the anterior ganglia of Aplysia. Previously, AVT, using similar methods, was found to be present only in vertebrates. AVT when perfused over the abdominal ganglion (10(-6)-10(-12)M) was found to increase the bursting activity of R15, to decrease the bursting activity of L3-L6 and to increase the CNS's suppressive influence over the gill withdrawal reflex evoked by siphon stimulation. The AVT present in the nervous system of Aplysia may mediate long-term suppression of gill reflex behaviors induced by factors such as satiation and, as well, regulate the activity of certain neurosecretory neurons. PMID- 7470891 TI - Spinal modulation of the acoustic startle response: the role of norepinephrine, serotonin and dopamine. PMID- 7470890 TI - Hypothalamic control of thermoregulation during dehydration. AB - Hypothalamic control of thermoregulatory evaporation has been examined in hydrated and dehydrated cats exposed to warm ambient temperatures. Reduction in evaporative cooling during dehydration is associated with increased hypothalamic temperature. Local thermal stimulation of the hypothalamus indicates that the thermosensitivity of hypothalamic systems controlling evaporation is reduced by dehydration. These findings are discussed in relation to control system models of thermoregulation. PMID- 7470892 TI - Biochemical evidence of decreased muscarinic cholinergic neuronal communication following amygdala-kindled seizures. PMID- 7470893 TI - Developmental changes in glycoproteins of the chick nervous system. AB - Temporal changes have been noted previously in retinal glycoproteins that bind to wheat germ agglutinin by a technique in which the denatured glycoproteins are first separated according to size by polyacrylamide gel electrophoresis, and are then localized on the gel using [125I]lectin. As reported here this technique will also detect differences between dorsal and ventral halves of the neural retina from 8-day chick embryos, and using other lectins will detect temporal changes in the glycoprotein pattern of the optic tectum. Some of the glycoproteins detected by wheat germ agglutinin in the neural retina appear to be represented on the surface of the retinal cells since: (a) the temporal changes in retinal glycoproteins can also be observed in a plasma membrane enriched fraction prepared from neural retina cells; and (b) antibodies prepared in mice against various size categories of wheat germ lectin binding glycoproteins bind to intact retinal cells. PMID- 7470894 TI - Impermeability of hagfish cerebral capillaries to radio-labelled polyethylene glycols and to microperoxidase. AB - The rate and extent of penetration of radiolabelled polyethylene glycols (900 and 4000 daltons) from plasma into brain was examined in the Atlantic hagfish (Myxine glutinosa), a primitive vertebrate, and, for comparison, in the rat. Tracer penetration into brain was limited to a similar extent in hagfish and in rats, indicating a basic similarity between the two species in the blood-brain barrier to polar non-electrolytes. The permeability of hagfish brain capillaries to the electron microscopic tracer microperoxidase (2000 Daltons) was also examined. Intravenously injected microperoxidase remained confined to the capillary lumen and was not found in the basal lamina or surrounding brain parenchyma. In contrast to an earlier study, these results indicate that the hagfish has a blood brain barrier to hydrophilic molecules larger than 900 Daltons. It is concluded that a blood-brain barrier is a general characteristic of all vertebrates. PMID- 7470896 TI - Contribution of noradrenaline-, dopamine- and adrenaline-containing axons to the innervation of different regions of the spinal cord of the cat. AB - The quantitative distributions of noradrenaline (NA), dopamine (DA) and adrenaline (A) were estimated throughout the grey and white matter at various levels of the cat spinal cord (C5, T3, T10 and L2). NA levels were high at all levels of the cord (about 1000 ng/100 mg protein), being more ventrally placed at the cervical level and tending to be more medially placed at lower thoracic and lumbar levels. DA was found at much lower concentrations than NA (less than 100 ng/100 mg protein) and generally decreased in more caudal segments. There was a mediodorsal distribution throughout the cord. A levels were extremely low throughout the cord (less than 17 ng/100 mg protein), but tended to be higher in the rostral-most segments. Up to 5 days after a total transection at T3, there was accumulation of NA in the white matter regions known to contain catecholamine axon tracts. DA, however, showed a much more restricted accumulation in the white matter. The possibility of whether each catecholamine exists separately as a neurotransmitter in the cat spinal cord is discussed. PMID- 7470895 TI - Modes of adaptation of peripheral neuroendocrine mechanisms of the sympatho adrenal system to short-term stress as studied in two inbred rat strains. AB - Alterations in mechanisms involved in catecholamine (CA) and corticosterone (B) regulation following short periods of stress have been investigated in two inbred rat strains: a strain more reactive to stress, Wistar-Kyoto (WK), and a less reactive strain, Brown-Norway (BN). Measurements after decapitation stress alone and after decapitation following different periods of immobilization stress, indicate that: (a) plasma CA levels immediately after severe stress (i.e. decapitation) are directly related to the behavioral reactivity of the two rat strains to stress but inversely related to adrenal gland size, their content of CA and biosynthetic enzymes, and to plasma levels of MHPG; (b) levels of B in adrenal glands and in plasma after stress are similar in both strains, in spite of different gland sizes; (c) CAT activity in presynaptic sympathetic terminals is directly related to plasma CA after short-term stress; and (d) liver COMT activity is directly related to plasma CA levels after stress, but inversely plasma levels of MHPG. The implications of the findings are discussed further in the text and lead to the conclusion that the BN strain represents a mode of a slower response to stressful stimuli than WK. PMID- 7470897 TI - Nucleus raphe dorsalis: a morphometric Golgi study in rats of three age groups. AB - Using Rapid Golgi and Nissl techniques, three major cell types: fusiform, multipolar and ovoid-shaped were identified in the nucleus raphe dorsalis of male rats at 30, 90, and 220 days of age. We have described the orientation and dendritic architecture of raphe cells as to type and the relationships of these cells to blood vessels and surrounding structures. For each cell type, the origin of the axon is characteristic. One hundred neurons per age group were measured at their maximal linear extent and the number of spines on the somal surface was counted. Dendritic number, linear extent, diameter and the number of spines along a 50 micron segment near the mid-point of dendritic length in an equal number of primary and secondary dendrites were quantified in each age group. The most striking age-related changes in the multipolar and ovoid-shaped cells were dendritic number, diameter and spine number as well as the number of perisomatic spines. The fusiform cells showed the least age-related changes. In general, the nucleus raphe dorsalis is organized as a reticular nucleus with neurons having few, straight and poorly ramified dendrites. PMID- 7470898 TI - Selective effects of LSD and hyperthermia on the synthesis of synaptic proteins and glycoproteins. AB - Protein synthesis in rabbit brain was inhibited following the intravenous injection of LSD. The incorporation of [35S]methionine into brain microsomal and synaptic fractions was decreased by 35-45% relative to control values. A selective increase was observed, however, in the relative labeling of a protein of molecular weight 75,000. Our previous studies have shown that LSD induces an increase in body temperature (i.e. hyperthermia) in rabbits. When LSD-induced hyperthermia was blocked the general reduction in labeling of microsomal and synaptic proteins was still apparent but the selective increase in relative labeling of the 75,000 dalton protein was not. Induction of hyperthermia by means other than LSD (i.e. elevation of ambient temperature) produced selective increases in the relative labeling of microsomal and synaptic proteins of molecular weight 75,000 and 95,000. These proteins are similar in molecular weight of two of the major 'heat shock' proteins whose synthesis is induced in several cultured cell lines following elevation of ambient temperature. Fractionation of [35S]methionine-labeled synaptic membranes by lectin affinity chromatography and analysis of [3H]fucose labeling patterns indicated that, in contrast to the general reduction in labeling of brain proteins, the synthesis of synaptic glycoproteins was not altered by LSD. The synthesis of glycosylated proteins present in other subcellular fractions was, however, reduced. These results suggest that LSD induced selective changes in the synthesis of brain proteins and that the synthesis of synaptic glycoproteins may be relatively resistant to drug administration. PMID- 7470899 TI - Different distribution of large and small retinal ganglion cells in the cat after HRP injections of single layers of the lateral geniculate body and the superior colliculus. AB - Retinal ganglion cells were labeled with HRP after injecting single layers of GL or single strata within the stratum griseum superficiale (SGS). Only small cells were labeled after injecting small cell C layers and upper SGS. Only large cells were labeled after injecting lower SGS. Small and large cells were labeled after injecting medial interlaminar nucleus (MIN) and layers A and A1. PMID- 7470900 TI - Thalamic reticular nucleus: anatomical evidence that cortico-reticular axons establish monosynaptic contact with reticulo-geniculate projection cells. AB - Cell bodies and dendrites of neurones in the thalamic reticular nucleus projecting to the dorsal lateral geniculate nucleus in adult albino rats, were labelled by retrograde transport of horseradish peroxidase (HRP). In the same animals the terminals of cortico-reticular axons were made identifiable by ipsilateral visual cortex ablation. Degenerating axon terminals established Gray type I synaptic contact with HRP-filled dendrites, indicating a monosynaptic connection between cortico-reticular and reticulo-geniculate projection cells. PMID- 7470901 TI - Isthmic afferent neurons identified by the retrograde HRP method in a teleost, Navodon modestus. AB - Isthmic afferent neurons were investigated by the retrograde horseradish peroxidase (HRP) method in a teleost, Navodon modestus. Following HRP injections into the nucleus isthmi, large pyriform neurons are labeled in the ipsilateral optic tectum. Very large and multipolar neurons are also labeled in the ipsilateral nucleus pretectalis. No labeled neurons were found in other areas. PMID- 7470902 TI - Descending inhibition of spinal neurons in the cardiopulmonary region by electrical stimulation of vagal afferent nerves. AB - Neurons in the left upper thoracic spinal cord of cats anesthetized with chloralose and relaxed with succinylcholine were activated by electrical stimulation of sympathetic chains or by pinching of the skin. Most neurons were spontaneously active, and 22 of the 51 neurons studied were antidromically activated from the spinomedullary junction, demonstrating their projection to higher centers. Stimulation of the central cut ends of either cervical vagus resulted in inhibition of firing in 26 neurons, acceleration in 6 neurons, both effects in 2 neurons, and no effect in the remaining 17 neurons. The parameters of stimulation determined whether inhibition of facilitation was produced in the two neurons that showed either effect. Two spinal neurons were fired by vagal stimulation with latencies of 16 and 20 msec. Activity in vagal afferent fibers is believed to impinge upon brain stem structures that have been shown by others to cause descending inhibition (and facilitation) of spinothalamic and spinoreticular neurons. This would provide a mechanism for modulation of sensory information from the cardiopulmonary region. PMID- 7470903 TI - Influence of 'strategy' on muscle activity during ballistic movements. AB - Human subjects were asked to grasp the handle of a manipulandum and to track a visual target with ballistic flexion or extension movements of the wrist. Prior to each trial subjects were given verbal instructions in order to influence their movement strategy. A burst of EMG activity was recorded from extensor carpi radialis (ECR) when subjects intended to terminate ballistic flexion movements within the target (antagonist burst). The antagonist burst in ECR was reduced or abolished during ballistic flexion movements when subjects intended to bang the manipulandum handle against a mechanical stop. Thus, our major observation is that antagonist muscle activity during ballistic movements is controlled by a subject's movement strategy. PMID- 7470904 TI - A barbiturate evoked intensification of postsynaptic inhibition in the preoptic ovulatory trigger region of the hypothalamus. AB - Extracellular single unit recordings were used to study inhibitory synaptic responses evoked from preoptic-anterior hypothalamic neurones following arcuate ventromedial stimulation. Intravenously administered methohexitone, pentobarbitone and thiopentone increased the duration of inhibitory synaptic responses by up to 400%. Submaximal responses to iontophoretically applied GABA but not glycine were also potentiated. Recovery from the actions of the short acting barbiturates was observed. PMID- 7470905 TI - Distribution of membrane glycoproteins among the organelles of a single identified neuron of Aplysia. II. Isolation and characterization of a glycoprotein associated with vesicles. AB - Glycoprotein-I (mol.wt. 180,000) is associated with a vesicle fraction from the cytoplasm of R2, the giant cholinergic neuron in the abdominal ganglion. Electron microscopy has shown that R2 contains both lucent and compound vesicles. We have used anisomycin, an inhibitor of protein synthesis, to provide evidence that Component-I is a constituent of somatic compound vesicles. In the presence of the drug, [3H]N-acetylgalactosamine injected into R2 is incorporated almost exclusively into Component-I. Quantitative electron microscopic radioautography of treated cells shows a marked increase in the proportion of silver grains over compound vesicles and a decrease in labeling of other organelles compared with untreated cells. Analysis of Component-I, isolated using the chaotropic agent lithium diiodosalicylate, shows it to contain fucose, N-acetylgalactosamine, and N-acetylglucosamine. Proteolytic digestion with pronase yields a complex pattern of glycopeptides. The proportion of [3H]N-acetyl-galactosamine in these glycopeptides is altered in the presence of anisomycin. These results together with radioautographic analyses suggest that large carbohydrate chains are elaborated within the endoplasmic reticulum and that smaller chains are added in the Golgi region or on the membrane of the compound vesicle. PMID- 7470906 TI - Pontine reticular unit responses to pelvic nerve and colonic mechanical stimulation in the dog. AB - The role of pontine reticula units (PRU) responding to afferent impulses from the distal colon and anal mucosa was studied in dogs. The 465 PRU were classified into 4 groups on the basis of the pattern of discharge in response to afferent stimulation (AS) of rectal branches (RB) of the pelvic nerve: Group I (G-I) responded with a short burst (latency, 36 +/- 16 msec); G-II responded with an early short burst (latency, 41 +/- 21 msec), followed by a late long-lasting train of spikes (latency, 187 +/- 110 msec); and G-III responded with a long lasting train of spikes (latency, 189 +/- 130 msec). G-IV had spontaneous discharges which were temporally inhibited by AS. G-II and G-III PRU showed remarkable facilitation similar to that of the long latency reflex discharges of RB. G-I PRU showed habituation. Most of G-II and G-III PRU discharged simultaneously with the reflex discharges of RB; these cause the contractions of defecation in the distal colon. In each group, 6.4 +/- 14.8% of PRU were reticulospinal units with descending axons running beyond the 1st lumbar segment of the spinal cord. It was concluded that the pontine defecation reflex center plays an essential role in defecating activity. PMID- 7470907 TI - Hippocampal serotonin re-uptake and nocturnal locomotor activity after microinjections of 5,7-DHT in the fornix-fimbria. AB - The role of the hippocampal 5-hydroxytryptamine (5-HT) terminals in the control of locomotor activity was investigated by lesioning 5-HT axons in the fimbria with 5,7-dihydroxytryptamine (5,7-DHT). Rats pretreated with desimipramine (10 mg/kg, i.p.) received microinjections of 5,7-DHT (0, 1, 3, 5 or 10 micrograms in 0.4 microliter ascorbic Ringer's solution) into the fornix-fimbria. On the fourteenth to twenty-first nights after operation, nocturnal locomotor activity was measured in photocell cages. Twenty-eight to thirty days after operation degeneration of 5-HT terminals was assessed by measuring in vitro [3H]5-HT re uptake in slices of dorsal hippocampus, ventral hippocampus and the septum. Groups injected with 5,7-DHT showed hyperactivity in the night period and increased decrements of activity between tests, both of which were related to the dose of neurotoxin. A reduction of [3H]5-HT re-uptake was found in dorsal hippocampus which was related to the dose of 5,7-DHT), but ventral hippocampal and septal [3H]5-HT re-uptake were not systematically reduced. For each rat, levels of dorsal and ventral hippocampal [3H]5-HT re-uptake were negatively correlated with the mean nocturnal activity from the 7 nights of testing. Levels of dorsal, but not ventral hippocampal [3H]5-HT re-uptake were negatively correlated with the mean nightly decrement of activity. No correlations were found between septal [3H]5-HT and these activity measures. These results, indicate that the increase in nocturnal locomotor activity caused by generalized depletion of 5-HT in the brain may be due to disruption of hippocampal 5-HT terminals supplied by the fornix-fimbria. PMID- 7470908 TI - An ultrastructural study of the synaptic contacts of alpha-motoneurone axon collaterals. I. Contacts in lamina IX and with identified alpha-motoneurone dendrites in lamina VII. AB - Horseradish peroxidase (HRP) was injected intracellularly in triceps surae alpha motoneurones. The axons and axon collaterals of these neurones were traced light and electron microscopically. Synaptic boutons of collaterals, in Rexed's lamina IX or in synaptic contact with HRP-stained motoneurone dendrites in lamina VII, were studied ultrastructurally. The boutons exhibited spherical synaptic vesicles and made synaptic contacts of two different types with HRP-stained alpha motoneurone dendrites in lamina IX and VII, dendrites and cell bodies of large neurones in lamina IX, dendrites of unknown origin in lamina IX and with one cell body of a medium size neurone in lamina IX. The observations are discussed in relation to earlier qualitative and quantitative studies on the synaptology of cat spinal alpha-motoneurones. PMID- 7470909 TI - The pretectal complex in the opossum: projections from the striate cortex and correlation with retinal terminal fields. AB - Two terminal fields were revealed in the pretectal complex of the opossum by the Fink-Heimer method after striate cortical lesions. A rostral field is located within a rostrolateral strip of the compact part of the anterior pretectal nucleus, where a partial topographic arrangement of this projection is present. A caudal field is located within the sub-brachial nucleus of the optic tract, located between the brachium of the superior colliculus and the posterior pretectal nucleus. The corticotopic projection to this field is mirror-symmetric to that found in the superior colliculus and overlaps a bilateral projection from the retina. Based on neural pathway evidence, it is concluded that the nucleus of the optic tract in the opossum can be subdivided in (a) an intrabrachial nucleus receiving a direct projection from the contralateral retina and (b) a sub brachial nucleus receiving projections from both retinae and from the striate cortex. The pretectal complex, as the superior colliculus, can be anatomically subdivided in a superficial region receiving visual input (the optic pretectum) and a deep region only remotely connected to the visual system. The optic pretectum, however, differs from the superior colliculus in displaying a multiple map arrangement within its constituent nuclei, instead of a single continuous representation of the visual field. PMID- 7470911 TI - Contralateral hindlimb responses to cutaneous stimulation during locomotion in high decerebrate cats. AB - High intensity stimuli of the skin were delivered to one hindlimb in various parts of the step cycle of decerebrate cats walking on a treadmill. Whereas the stimulated limb always flexes after the stimulation there is, in the contralateral limb, a crossed flexion response during swing or a crossed extension response during stance. The frequency distribution of the responses in contralateral flexor or extensor muscles peaks at around the onset of the respective locomotor bursts although responses can be evoked before and after that onset. In certain periods of the cycle, largely corresponding to the transition from flexor or extensor activity and vice versa, the responses can occur in either muscles. The long latencies of crossed responses are similar to that of the ipsilateral responses so that both occur approximately at the same time. The amplitude and duration of crossed responses vary according to the phase of the walking cycle. The types of responses and the overall changes in the step cycles suggest that crossed extension responses mainly serve to sustain the increased weight to the contralateral side during ipsilateral flexion whereas crossed flexion responses appear well studied to rapidly rephase the contralateral limb step cycle to that of the ipsilateral limb which is markedly perturbed by the flexion response elicited during its stance phase. PMID- 7470910 TI - The role of the midbrain in courtship behavior of the female ring dove (Streptopelia risoria): evidence from radiofrequency lesion and hormone implant studies. AB - The involvement of the nucleus intercollicularis (ICo) region of the midbrain, an estrogen-sensitive area, in the expression of estrogen-dependent female courtship behavior was examined with radiofrequency lesion and intracranial hormone implant techniques. Bilateral lesions in the ICo region caused a reduction in the female's nest coos, and no reduction in other behaviors in response to male courtship. In addition, the follicles of the ICo lesioned females failed to grow in response to male courtship. A second experiment showed that the difference in follicular size between ICo-lesioned and sham-ICo-lesioned females was not observed if females were visually isolated from males, suggesting that tonic ovarian activity was not affected by ICo lesions. In the third experiment, unilateral 30-gauge implant containing estrogen (E) or estradiol benzoate (EB) in the ICo region of bilaterally ovariectomized females restored the nest-coo, though not to the level obtained with systemic EB injections. Other behaviors were not selectively affected by E or EB implants in the ICo region. Diffusion from the implant was probably not responsible for the elicitation of the nest coo, since the oviduct weights of females implanted with E or EB and cholesterol did not differ. These data are consistent with the hypothesis that the ICo region is involved in the expression of vocal (cooing) courtship behavior in the female ring dove. PMID- 7470912 TI - Midbrain dopamine neurons: differential responses to amphetamine isomers. AB - Intravenously administered D- and L-amphetamine have different potency ratios in reducing the firing rates of dopamine cells in the substantia nigra and in the ventral tegmental area. While D-amphetamine is considerably more potent than L amphetamine in reducing ventral substantia nigra dopamine neuronal impulse flow, D- and L-amphetamine are of similar potency in reducing dorsal substantia nigra and ventral tegmental dopamine neuronal impulse flow. These results suggest that all dopamine cell groups are not pharmacologically identical and that different dopamine nuclei may respond differently to psychoactive drugs. The comparable potencies of the D- and L-isomers on dorsal substantia nigra and ventral tegmental area dopamine neurons may explain, by a dopamine mechanism, the finding that comparable doses of the isomers produce schizophrenic-like symptoms. PMID- 7470913 TI - Afferent projections to the ventral lateral geniculate nucleus in the cat. AB - Horseradish peroxidase (HRP) injections were made into the dorsal lateral geniculate nucleus (LGNd) and ventral lateral geniculate nucleus (LGNv) of the cat in order to define afferent projections to LGNv. These were found from the superior colliculus, contralateral LGNv, dorsal median raphe nucleus, locus coeruleus, ipsilateral pretectum, and various portions of visual cortex. While many cortical areas project to LGNv (17, 18, 19, 21 and lateral suprasylvian), the heaviest input arises from areas 17 and 20. The cell bodies of origin are in layer 5 in contrast to layer 6 which projects to LGNd. PMID- 7470914 TI - Growth of sympathetic nerve terminals in the adult rat. AB - This report presents evidence that in adult life, in the absence of any experimentally-induced stimulus, the density of sympathetic nerve terminals increases in rat submaxillary salivary glands, suggesting that sympathetic nerve terminal growth continues in adult life. Several theories which have been proposed to account for nerve terminal growth in adult animals are discussed in light of experimental results. PMID- 7470915 TI - In vivo voltammetric investigations into the action of HA-966 on central dopaminergic neurons. PMID- 7470916 TI - Batrachotoxin induced axonal necrosis in peripheral nerves. AB - Batrachotoxin (BTX) depolarizes electrogenic membranes and also blocks axonal transport. We demonstrate that when applied in doses sufficient to block impulse transmission and axonal transport, BTX induces axonal necorsis. It is postulated that axonal necrosis is secondary to influx of sodium ions and water. PMID- 7470917 TI - Increased hypothalamic norepinephrine metabolism after water deprivation in the rat. AB - Rat brain catecholamine metabolism was changed over a period of several days by limited access to water (10 min/day). One or two weeks limited access to water caused an increase in hypothalamic norepinephrine metabolism as measured with alpha-methyl-para-tyrosine. Brain stem and telencephalon norepinephrine was not affected by the limited access to water regimen. Dopamine metabolism in the corpus striatum and the hypothalamus was not altered by limited access to water. If the limited access to water was continued for 3 or more weeks, hypothalamic norepinephrine metabolism then returned to normal. The increase in hypothalamic norepinephrine metabolism was confirmed by a second method measuring in vivo tyrosine hydroxylase activity. Additional experiments demonstrate that this affect is specific for water deficits. Limited access to food had no effect on the metabolism of norepinephrine in the hypothalamus. Water deficits produced by replacing water with a 2% NaCl solution caused a similar increase in hypothalamic norepinephrine metabolism to that observed after one week limited access to water. Furthermore, 10 min access to water stopped the increased hypothalamic metabolism of norepinephrine seen after one week of limited access to water. The regional specificity (effect seen in hypothalamus but not the telencephalon and brain stem), and the stimulus specificity (water and not food deficits) suggest hypothalamic norepinephrine involvement in thirst or hormonal control of water regulation. PMID- 7470918 TI - Morphological changes in rat brain induced by L-cysteine injection in newborn animals. AB - A single subcutaneous injection of L-cysteine (1.2 mg/g body weight) to rats 4 days after birth was followed by atrophy of the brain which was well developed 27 -32 days after the injection. It was apparent that the lesioned animals could be divided into two groups (type 1 and 2) on account of the degree of brain atrophy. In type 1, which was observed in 80% of the animals, the body weight was unchanged, but the total brain weight was reduced by about 20%. The brain structures most affected were cerebral cortex, hippocampus and thalamus, each having a 30--40% reduction in wet weight. The atrophy of the posterior part of cortex was particularly pronounced in this type of lesion. In type 2 lesion, which appeared in 10% of the survivors, the atrophy was much more severe. There was a 50% reduction in wet weight of brain and in body weight. The most prominent finding was the atrophy of the whole cortex and the hippocampus which were reduced by 80 and 60% of wet weights respectively. In this type of lesion significant morphological changes were observed in several brain regions such as caudato-putamen, thalamus, pons, medulla oblongata, spinal cord and cerebellum. PMID- 7470919 TI - The organization of visceral sensory neurons in thoracic dorsal root ganglia (DRG) of the cat studied by horseradish peroxidase (HRP) reaction using the cryostat. AB - The organization of visceral sensory neurons in thoracic dorsal root ganglia (DRG) was studied by retrograde transport of horseradish peroxidase (HRP) from the central cut end of the left major splanchnic nerve of the cat. The majority of HRP-labeled cells were concentrated between T5 and T11. Within a DRG, labeled splanchnic neurons were found in all sectors. There was no consistent pattern of localization within the ganglion although clustering of visceral cell bodies was apparent. It may be that each clustered group of cells innervates individual viscera or reflects a degree of functional segregation. PMID- 7470920 TI - Freeze-fracture of developing neuronal plasma membrane in postnatal cerebellum. AB - The structure of the plasma membrane of two neuronal cell types was quantitatively assessed in freeze-fracture replicas of rat cerebellar cortex from 2 to 73 postnatal days. The measurement of the number and size of intramembrane particles (IMP) in different regions of the Purkinje cells and of the granule cell showed that between 2 and 21 postnatal days there was a differential increase in the number of IMP; several regions of the cell characterized by a different pattern of synaptic interconnections showed differences in particle size and in particle number: neuronal processes had lower numbers of IMP than perikarya, whereas dendrite spines contained fewer IMP than dendritic trunks. These differences in IMP content appeared at the time at which specific synaptic contacts are established in these various regions of the cells. PMID- 7470921 TI - Pressor effects of dorsal raphe stimulation and intrahypothalamic application of serotonin in the spontaneously hypertensive rat. AB - Electrical stimulation of the dorsal raphe nucleus or direct microinjection of serotonin into the preoptic region of the anterior hypothalamus produces a transient rise in arterial blood pressure in both spontaneously hypertensive rats (SRH) and Wistar--Kyoto (WKY) controls. SRHs are more responsive to raphe stimulation but are somewhat less responsive to serotonin injections when compared to WKYs. The serotonin antagonist metergoline blocks the pressor response to serotonin in both strains. These results suggest that the central serotonergic neuronal system play a similar, but not identical, role in blood pressure modulation in hypertensive and normotensive rats. PMID- 7470922 TI - Plasticity of binaural interaction in the cat inferior colliculus. AB - Responses of single neurones in the inferior colliculus (IC) to acoustic interaural intensity difference (IID) were examined in normal, adult cats and in cats that had been reared for 3--4 months, either from birth or as adults, with unilateral ligation of the external meatus. There were significantly fewer units displaying IID sensitivity in either of the ligated groups than there were in the normal group. The loss of IID sensitivity in the ligated animals reflected a diminished inhibitory input from the non-ligated ear. PMID- 7470923 TI - The influence of eccentricity on receptive field types and orientation selectivity in areas 17 and 18 of the cat. PMID- 7470924 TI - Age-dependent changes in the beta-endorphin content of discrete rat brain nuclei. AB - Concentrations of beta-endorphin were measured by radioimmunoassay in discrete brain nuclei of young (3 months) and old (24 months) rats. The beta-endorphin content of all major structures known to contain this peptide with the exception of the eminence, is reduced (50.3 +/- 2.8% of control) in old rats. PMID- 7470925 TI - Presynaptic regulation of the release of catecholamines in the cat hypothalamus. AB - The posterior hypothalamus of cats was superfused through a push-pull cannula and the release of endogenous catecholamines was determined in the superfusate. Superfusion with yohimbine, isoprenaline, salbutamol or tazolol increased, while superfusion with propranolol decreased, the release of all three catecholamines. Transection of the brain caudal to the hypothalamus inhibited 'resting' and drug induced release. It is concluded that alpha- and beta-adrenoceptors of the hypothalamus are involved in the regulation of the release of catecholamines. PMID- 7470926 TI - Stumbler, a new mutant mouse with cerebellar disease. AB - A new mutant mouse named Stumbler (stu) displays clinical features suggesting a cerebellar lesion. The main light microscopic findings, based on a Golgi technique and on sections of plastic embedded material, are that Purkinje cells in the mutant cerebellum have small dendritic arborizations and exhibit immature spines on their somata. Purkinje cells also contain an increased number of mitochondrial profiles both in cell bodies and in swellings on dendrites. PMID- 7470927 TI - Quantitative capillary topography and blood flow in the cerebral cortex of cats: an in vivo microscopic study. AB - In 50 anesthetized cats the microcirculation in intermediate and deeper layers of the cerebral cortex was visualized in vivo by microtransillumination, and documented by high-speed microcinephotography. The viability of the preparation was verified in a series of experiments demonstrating spontaneous vasomotion and responsiveness to chemical stimulation of pial arterioles and small arteries. Stereological methods for quantitative analysis of projected images of capillaries in a comparatively large tissue volume were employed to determine morphometric and topographical parameters of the asymmetric, highly tortuous intracortical capillary network. Capillary diameters (5.1 +/- 0.84 micrometer), radii of curvature (median 57 micrometer), total capillary lengths per tissue volume 939 +/- 338.2 mm/cu.mm), capillary volume fractions (2.1 +/- 0.51%), total capillary surface areas per tissue volume (15.3 +/- 4.85 sq.mm/cu.mm), and intercapillary distances (median 24.2 micrometer) showed significant interregional differences. The frequency distribution of the lengths of capillary segments (median 108 micrometer) was best described by a Weibull distribution. On the average 90% of all capillaries were continuously perfused. Capillary red cell flow (median velocity 1500 micrometer/sec) was predominantly unidirectional and conspicuously irregular. The variance of capillary red cell velocities (CRCVs) was significantly correlated (tau = 0.48) with capillary tortuosity. An extreme value distribution best described the observed frequency distribution of CRCVs. Flow irregularities represented both white noise and a significant stochastic periodicity at frequencies between 40 and 90 Hz. PMID- 7470928 TI - The influence of protein restriction, rehabilitation and changing nutritional status on synaptic development: a quantitative study in rat brain. AB - Quantitative ultrastructural technique were employed to compare the development and maturation of cortical synapses in rats subjected to protein deprivation, control diet, nutritional rehabilitation and a normal-low crossover diet. Osmicated preparations of the molecular layer of occipital cortex were prepared at 15, 20, 28, 75 and 224 days postnatal in male rats. At 15 and 30 days the values for mitochondrial and synaptic densities, vesicle number and packing per terminal, synaptic length, terminal area and brain weight were lower in the protein-deprived than in the control animals. These initial deficits were progressively transformed to excesses, with the most marked crossover period occurring between 20 and 28 days. By 224 days only brain weight and presynaptic terminal area were lower in the protein-deprived material. A further parameter, synaptic curvature, showed a decrease in negativity in protein-deficient junctions between 15 days (64%) and 224 days (41%). Well-nourished tissue had fewer negatively-curved synapses during early development. These data suggest that the morphological development of the presynaptic terminals is not simply delayed, but is ultimately different in the protein-deprived animals. The nature of these changes may facilitate an adaptation to provide more efficient functioning in the adverse condition. Nutritionally rehabilitated tissue ultimately approaches the control, although significant differences occur in synaptic and mitochondrial densities and in brain weight. The normal--low crossover procedure intensities the differences noted in animals subjected to continuous deprivation. PMID- 7470929 TI - Central correlates of peripheral pattern alterations in the trigeminal system of the rat. II. The effect of nerve section. AB - The effect of trigeminal nerve section on pattern formation in the rat central nervous system was investigated. Nerve section was found to abolish all evidence of afferent segmentation in the brain stem. At thalamic and cortical levels the pattern of afferent segmentation was affected in a time-dependent fashion. The later the procedure is performed, the less severe the resultant effect. Early section abolishes all afferent segmentation, later section only within-row segmentation, and still later section has no effect on pattern formation. The results of this experiment support the hypothesis that peripheral spatial information plays an important role in pattern formation in the central trigeminal system. PMID- 7470930 TI - Stroboscopic rearing reduces direction selectivity in rabbit visual cortex. AB - Rabbits reared from birth in stroboscopic illumination have no experience of visual motion. In primary visual cortex of these animals there is a large reduction in the number of cells which are direction selective. This result, contrary to previous reports, shows that the rabbit visual system can be modified by early visual experience. PMID- 7470932 TI - Abnormal maturation of cerebral cortex and behavioral deficit in adult rats after neonatal administration of antibodies to ganglioside. AB - Five-day-old rats received a single injection (50 microliter) of antiserum to ganglioside into the cisterna magna and were compared to control animals injected with the antiserum which had been absorbed with pure GM1 ganglioside to remove the specific antibodies. Both groups showed normal rates of body growth. However, animals receiving antiganglioside serum had, at 60 days of age, impaired performance when tested on a complex learning task (DRL) as well as chemical and morphological alterations in the somatosensory cerebral cortex. Gross morphology and wet weight of whole brain were normal in both groups. Microchemical analysis of somatosensory isocortex revealed a normal content of total solids, protein, and DNA. However, ganglioside sialic acid, galactocerebroside, and RNA were decreased by 31%, 32% and 25% of control values, respectively (P less than 0.01). Quantitative measurements of oblique dendrites of Golgi-stained cortical pyramidal neurons revealed a 31% decrease in the number of spines. Additionally, the majority of spines were of the stubby configuration, whereas dendrites from controls were populated predominately by thin spines. These observations suggest that antibodies to GM1 ganglioside interfere with optimal neonatal development on both dendrites and myelin in cerebral cortex. The results provide an animal model in which an immunologically-mediated disturbance of cortical development is associated with chronic behavioral impairment. PMID- 7470931 TI - Strabismus disrupts binocular convergence in cat nucleus of the optic tract. AB - Rendering kittens strabismic by surgical section of the medial rectus muscles results in disruption of binocular convergence in the nucleus of the optic tract. Visual input through the ipsilateral eye of strabismic cats does not influence cells in this nucleus although nearly one half of the cells encountered in normal cats receive inputs from both eyes. The results provide a basis for the deficits in optokinetic nystagmus found in strabismic cats and suggest a new mechanism underlying competition between inputs from the two eyes during postnatal development. PMID- 7470933 TI - Postnatal development of ganglionic neurons in the absence of preganglionic input: morphological observations on synapse formation. AB - Synapse formation during postnatal development of the superior cervical sympathetic ganglion was studied in rats after neonatal ganglionic denervation, when reinnervation was either permitted or prevented. In both groups of operated animals, 90% of the synapses were lost by the fourth postoperative day. In the rats in which reinnervation was permitted, restoration of synapse numbers began by one month after surgery and reached 50% of control by two months. There was no further synapse restoration after this time. In the animals in which reinnervation was prevented, synapse numbers increased, but at all times were approximately 10% of controls. Thus the intrinsic ganglionic synapses underwent their normal postnatal developmental increase in number, but did not sprout to any significant degree in response to the massive deafferentation caused by removal of the preganglionic input. Vacant postsynaptic membrane thickenings (those not apposed by presynaptic terminals) appeared in both experimental groups. There was no significant loss of these vacant thickenings in either group over the course of 3 months. Therefore, the vacated postsynaptic sites do not appear to be recontacted during reinnervation, while the reinnervating axons appear to cause the formation of new postsynaptic sites on the ganglion cells. PMID- 7470934 TI - Fate of the multiple innervation of cerebellar Purkinje cells by climbing fibers in immature control, x-irradiated and hypothyroid rats. AB - The fate of the multiple innervation of Purkinje cells (PCs) by climbing fibers (CFs) was studied as a function of age in immature rats rendered agranular by X irradiation, in immature hypothyroid rats, and compared to that in controls. This was done by examining in each group the intracellular activities of PCs mediated via CFs throughout maturation. From the third day in control rats, CF responses of PCs evoked by juxta fastigial region (JF) stimulation or occurring spontaneously already resembled the adult responses with, however, some important differences: (1) most of these responses were graded by steps with the intensity of the stimulation before day 13, due to the multiple innervation of PCs by CFs (see below); (2) immature CF responses exhibited a longer duration and their initial spike started near the peak of the EPSP instead of near the baseline later on. Finally, an anlage of CF response was already present in most PCs on day 2, and consisted of a single fast spike elicited near the peak of an underlying all-or-none EPSP. In the 3 groups of rats, CF EPSPs already closely resembled the adult ones as early as 3 days, although their total duration and especially their time to peak were longer. In control rats, these CF EPSPs reversed with depolarizing currents from day 3 and currents for reversal were much lower than in the adult. 'Dual' CF EPSPs of PCS37 were encountered in immature 7- to 10-day-old controls, and persisted in hypothyroid rats until the end of the third postnatal week. The mono- or the multiple innervation of PCs by CFs was ascertained in th 3 groups according to the graded or the all-or-none character of CF EPSPs, and the number of CFs impinging on a given PC was estimated by the number of steps in the response. In control rats, most of PCs were already multiply innervated by CFs as early as 3 days. The multiple innervation culminated on day 5 with an average number of 3.4 CFs for PC, and rapidly regressed later on, so that the adult-type monoinnervation was the rule after day 13. In hypothyroid rats, the establishment of the redundancy and its regression was delayed by 2--3 days. In X-irradiated rats, the settlement and the involution of the multiple innervation of PCs by CFs was exactly superimposed with that seen in controls until day 8. Later on, regression of the supernumerary contacts no longer occurred so that most PCs remained multiply innervated until adulthood. Finally, the first clear-cut IPSPs were detected in PCs on day 9 in control and X-irradiated rats and 2--3 days later in hypothyroid animals. PMID- 7470935 TI - Trophic activities for dorsal root and sympathetic ganglionic neurons in media conditioned by Schwann and other peripheral cells. AB - We describe here the use of 3 established nerve growth factor (NGF) targets (cultured neurons from dissociated chick embryo sympathetic ganglia, and chick embryo or neonatal mouse dorsal root ganglia) to investigate neuronotrophic activities in conditioned media (CMs) from: (i) chick embryo heart; (ii) purified mouse Schwann cells; and (iii) clonal rat Schwannoma RN22 cells. In chick sympathetic and mouse dorsal root ganglionic cultures, all 3 CMs supported survival of the same number of neurons as did mouse submaxillary NGF, and in most cases no increased survival resulted from concurrent administration of NGF and any one CM. NGF and CM activities were quantitated in each of the responsive cell systems. No differences were seen when either test population was used for the same agent, or when different CMs were examined on the same test cells. The CM activity, unlike that of NGF, was not blocked by even excess amounts of antiserum against mouse submaxillary NGF. The neuronotrophic activity of CMs appears to reside with macromolecular constituents. None of the CMs displayed trophic activity on chick embryo dorsal root ganglionic neurons. However, at least one of them (RN22 medium) had drastic effects on these ganglionic cells even in the presence of NGF, leaving open the possibility of a 'toxic' factor overriding putative trophic agents. PMID- 7470936 TI - Long term potentiation of lateral hypothalamic self-stimulation following parabrachial lesions in the rat. AB - Two neural systems in the pontine tegmentum have been implicated in reward processes. One is the visceral afferent-column arising from the nucleus of the solitary tract and the pontine tegmental taste area (PTA), the other being the dorsal noradrenergic system that has its origin in the locus coeruleus (LC). Lesions of the LC had no effect on lateral hypothalamic (LH) intracranial self stimulation (ICSS) whereas lesions of the PTA produce a dramatic and seemingly permanent increase in LH ICSS response rates. These results are discussed in the context of noradrenergic/dopaminergic interactions in the control of goal directed behaviors. PMID- 7470937 TI - The role of protocerebrum in the modulation of circadian rhythmicity in the crayfish visual system. AB - Dark-adapted crayfishes with protocerebrum only, were submitted to continuous recordings of electroretinogram (ERG) and of eye glow area (EGA) during several days. Circadian variations of ERG amplitude similar to that of intact animals, were revealed by means of restrained test light stimuli (0.2 Cd/ft2) bilaterally applied to each eyestalk. The period (24.6-38 hr) and range (40-80%) value of ERG oscillations always resulted quite similar to one another side. As in intact animals retinal shielding pigments (RSP) position as measured as EGA size showed a clear circadian rhythm, and also a clear consensual reflex in these preparations. We found a loss of both: circadian and consensual mobilization of distal RSP in animals with complete removal of cerebral ganglion. Our proposition is that the crayfish protocerebrum plays a major role in the modulation of circadian retinal sensitivity, probably through the control-release of hormonal neurosecretions from the sinus gland along the day. PMID- 7470938 TI - Malformation of Purkinje cell dendrites induced by graft-versus-host disease. AB - Impairment of cerebral Purkinje cell growth was assessed in Golgi-Cox stained tissue in 14 day old (Fischer X DA)F1 hybrid rats subjected to graft versus host disease (GVHD). GVHD was induced by grafting parental strain lymph node cells (PSLNC) into the anterior facial vein on the day of birth. We have previously described GVHD induced changes in nucleotide and protein content [12] and RNA function [14] as well as alterations in cell numbers and areas of the external granular and molecular layers in diseased animals [13]. In this report some effects of GVHD on the gross morphology of Purkinje cells in 14 day old animals are enumerated. Most Purkinje cells (62%) from GVHD animals had a height to width ratio greater than 1, whereas most from control animals (71%) had ratios of less than 1. The majority of Purkinje cells (67%) from diseased animals had elongated main dendrites which were devoid of branches and often (13%) these elongated main dendrites were S-shaped. In addition, comparison of Purkinje cells from GVHD and control animals revealed a greater tendency (15%) toward retention of extrasomal process in GVHD animals. These findings suggest that GVHD affects nondividing, differentiating cell populations as well as those which are proliferating and migrating. Our evidence that, as a result of GVHD, the protein synthesizing capacity of the cerebellum is altered [14] and that Purkinje cells are more closely spaced at day 11 suggests that both intrinsic and extrinsic factors are involved in producing the changes in dendritic shape reported here. PMID- 7470939 TI - Changes in rat brain norepinephrine levels and turnover after olfactory bulbectomy. AB - After bilateral olfactory bulbectomy in rats a significant increase of norepinephrine (NE) level in the hypothalamus was found. However, no difference was observed between hypothalamic NE turnover of bulbectomized and sham operated animals in the amygdaloid cortex the NE level was not affected by bulbectomy. In this area, however, the NE turnover appeared to be decreased after bulbectomy. The latter finding may be related to the deficits in passive avoidance behaviour as found in bulbectomized rats. PMID- 7470940 TI - The conduction velocity and central projections of retinofugal fibers in the rabbit. AB - IN anesthetized and paralyzed rabbits, action potentials were elicited antidromically following electrical stimulation of optic tract terminals at the geniculate level (LGN). The conduction velocity spectrum extended from 7 m.s-1 to 34 m.s-1. The distribution of conduction velocities indicated four major modes at 10, 18, 22 and 26 m.s-1. Antidromic compound action potentials exhibited good correlation between the conduction latency and the major modes of the distribution histogram. These results suggest that the rabbit optic tract is composed of four classes of fibers varying in their conduction velocities. The central projections of retinofugal axons were studied with electrical stimulations of the Superior Colliculus (CS) and the LGN while recording from the same optic tract fiber. Antidromic spikes could be elicited from all conduction velocity groups and 71% of axons responded to both sites of stimulation. This finding indicates that most retinofugal fibers branch to and innervate both CS and LGN. Further, there is a tendency for fast-conducting axons to have their receptive fields located eccentrically relative to the optic axis of the eye. PMID- 7470941 TI - Biochemical characterization of solubilized muscarinic acetylcholine receptors. AB - Several in part new methods (salt extraction, phospholipase treatment, transfer to serumlipoproteins, nonionic detergent mixtures) have been extensively examined in order to obtain a solubilized muscarinic acetylcholine receptor with properties suitable for further purification. An optimal mixture of digitonin and gitonin has been found which forms a tightly bound detergent wall around the receptor protein as indicated by ion exchange and monolayer studies and which allows to solubilize the receptor from enriched synaptosomal membranes with a high yield and stability. The specific 3H-QNB binding of this preparation is nearly unchanged between pH 7.5 and 10. The isoelectric point of the muscarinic acetylcholine receptor has been determined by isoelectric focusing to be between pH 4.5 and 4.6. PMID- 7470942 TI - Intracellular analysis of synaptic potentials in rat neostriatum following stimulation of the cerebral cortex, thalamus, and substantia nigra. AB - Intracellular recordings were obtained from neostriatal neurons of unparalyzed male hooded rats anesthetized with urethane. Electrical stimulation of the cerebral cortex (Cx), centromedian-parafascicular area of the thalamus (CMP), and the substantia nigra (SN) elicited monosynaptic excitatory postsynaptic potentials (EPSPs) in neostriatal neurons. Response latencies were, on the average, 3.7 msec, 3.3 msec, and 3.8 msec, for Cx, CMP and SN stimulation, respectively. Over 85% of recorded neurons showed convergence of inputs from all three stimulation sites. The SN induced EPSP sometimes had two components, with the second component beginning 10-15 msec after the first. EPSPs from all three stimulation sites were often followed by inhibitory postsynaptic potentials (IPSPs) lasting from 50-250 msec. Double shock experiments indicated that SN induced EPSPs could be reduced in amplitude by 20-80% when preceded by conditioning stimulation to Cx, CMP or SN. In contrast, the EPSP elicited by Cx stimulation were unaffected by conditioning stimulation. Some recorded neurons were morphologically identified by means of intracellular injection of horseradish peroxidase. All were "medium spiny" neurons. The results of the present study agree well with those of previous studies of cat caudate neurons, and extend them to rat neostriatal neurons. PMID- 7470944 TI - Electroacupuncture suppression of the rat jaw opening response after stimulating the caudal spinal trigeminal nucleus. AB - Suppressive effects of electroacupuncture on the jaw opening responses evoked by stimulation of two areas (anterior and posterior) within the caudal spinal trigeminal nucleus were investigated using lightly anesthetized rats. Electroacupuncture stimulation markedly suppressed the jaw opening responses evoked by stimulating the posterior area, but suppressed only slightly those evoked by stimulating the anterior area. PMID- 7470943 TI - The role of protein synthesis in the hypothalamic mechanism mediating pyrogen fever. AB - The effects of inhibition of protein synthesis by anisomycin on the pathogenesis of fever and normal thermoregulatory processes were investigated in the conscious and unrestrained cat. Subcutaneous administration of 5.0-25.0 mg/kg of anisomycin prevented the fever normally evoked by an intravenous infusion of either 1.0 ml (10(8) organisms) of a 1:10 dilution of S. typhosa or 1.0-5.0 ml (3.5 x 10(5)-2.1 X 10(7) cells/ml) of endogenous pyrogen. In addition, systemic pre-treatment with anisomycin delayed and/or blocked the fever typically elicited by a direct micro injection into the anterior hypothalamic, preoptic area (AH/POA) at AP 12.5-16.0 of 1.0 microliters of the endotoxin. Anisomycin did not alter the hyperthermic response to an anterior hypothalamic injection of either 1.0-7.0 micrograms/1.0 microliters of serotonin (5-HT) or 100.0 ng/1.0 microliters of prostaglandin (PGE). Inhibition of protein synthesis, furthermore, did not prevent the fall in body temperature usually produced by an intrahypothalamic micro-injection of 2.33 14.0 micrograms/1.0 microliters of either norepinephrine (NE) or dopamine (DA). The thermoregulatory capacity of the cat was unaffected by the administration of comparable doses of anisomycin, i.e., the animal was able to maintain normal body temperature (+/- 0.5 degrees C) when exposed to an ambient temperature of either 10 degrees C or 34 degrees C. These results strongly suggest that the synthesis of new protein within the region of the AH/POA is a functional requisite for the development of a pyrogen-induced fever. PMID- 7470945 TI - An inexpensive electronic switch for preventing amplifier overload in neurophysiological recording. AB - A circuit for an electronic switch is described that suppresses large unwanted signals that might overload physiological amplifiers during recording of weak neural signals. The circuit, made from inexpensive and easily available integrated circuits, can be operated from any timer that produces at least a +5 volt output pulse. It operates by connecting a low gain preamplifier to a high gain amplifier only during the signal of interest. PMID- 7470946 TI - Modification of rabbit head holder to facilitate stereotaxic brain surgery. AB - The Narashige stereotaxic headholder for the rabbit is modified by attaching a graduated scale and pointer on the metal arc of the machine. This modification greatly simplifies setting of the horizontal zero plane. PMID- 7470948 TI - The origin of forebrain afferents to the habenula in rat, cat and monkey. AB - Injections of horseradish peroxidase (HRP) involving the entire habenular complex in rat, cat and squirrel monkey (Saimiri sciureus) label (1) numerous cells in anterior lateral hypothalamic area, (2) a moderate number of cells in lateral preoptic area, substantia innominata, nucleus of diagonal band and postcommissural septum, and (3) a few cells in medial hypothalamus, ipsilaterally, in all three species. Some labeled cells also occur in corresponding regions contralaterally. The contribution of these limbic structures to the innervation of habenula is thus strikingly similar in the three groups. In contrast, significant species variations are found in respect to pallidal afferents. Whereas the entopeduncular nucleus in rat stands out as the main source of forebrain habenular afferents, the same structure in cat appears to contribute less substantially than adjoining lateral hypothalamus to the innervation of habenula. In monkey habenular afferents also arise principally from lateral hypothalamic neurons. At pallidal levels, labeled cells are nevertheless abundant in the rostral pole of primate internal pallidum. More caudally, they are found in significant number along internal and accessory medullary laminae where they intermingle with acetylcholinesterase-containing neurons which do not themselves project significantly upon habenula. This heterogeneous distribution of labeled pallidal cells indicates that the pallidohabenular projections in primate may arise, at least in part, from specific neuronal subpopulations within internal pallidum. PMID- 7470947 TI - Differential distribution of norepinephrine and serotonin along the dorsal ventral axis of the hippocampal formation. AB - Norepinephrine (NE) and Serotonin (5-HT) are differentially distributed along the dorsal-ventral axis of the Hippocampal Formation (HF). A kinetic analysis of the uptake of labeled NE and 5-HT was performed to assess the relative density of nerve terminals which accumulate NE and 5-HT (Vmax) and the apparent affinity for uptake (Km). The results reveal a greater density of NE and 5-HT terminals in the ventral HF and a higher Km for 5-HT uptake in the dorsal HF. The results are discussed in terms of potential explanation of reported differences following independent manipulation of the dorsal and ventral hippocampal formation. PMID- 7470949 TI - The descending afferent connections of the paraventricular nucleus of the hypothalamus (PVN). AB - The distribution of afferents to the paraventricular nucleus of the hypothalamus (PVN) was studied following iontophoresis of horseradish peroxidase (HRP) into the nucleus. This report describes the descending connections to this structure. The PVN receives a substantial input from limbic system structures, especially from the lateral septal nucleus and the ventral portion of the subicular cortex. A small number of labeled cells were found in the amygdala, primarily the medial nucleus. Of the circumventricular organs, the subfornical organ and OVLT both project to the PVN, the former very strongly. In both the preoptic area and hypothalamus, medial cell groups have more input than do the lateral areas. Labeled cells were found in the medial and hypothalamus, medial cell groups have more input than do the lateral areas. Labeled cells were found in the medial and lateral preoptic areas, suprachiasmatic nucleus, ventromedial nucleus, arcuate nucleus, retrochiasmatic and lateral hypothalamic areas. Of particular interest is the finding that the PVN receives input from the contralateral PVN and ipsilateral SON. Of the other diencephalic structures only the zone incerta showed a consistent number of labeled cells. The results are discussed in relationship to the possible neural structures that might mediate the response of the PVN neurons to adrenalectomy. PMID- 7470950 TI - Identical responses of the two hippocampal theta generators to physiological and pharmacological activation. AB - The possible existence of two separate RSA generators in the dorsal hippocampus raises questions as to the degree of coupling or independence of both. In order to answer these questions bipolar EEG recording were made in the CA1 and dentate gyrus of freely moving rats during conditions of RSA generation by different manipulations. The quantitative EEG parameters of simultaneous records were compared and cross correlation functions were calculated. Results indicate that even under different conditions of inducing RSA (physiological activation or injection of arecoline and nicotine) yielding RSA with different frequency characteristics, the quantitative EEG parameters were similar in both recordings. In addition, a high degree of concordance of both EEG was evidenced in the cross correlation function. These observations are discussed in terms of a high degree of coupling between both RS generators. PMID- 7470951 TI - Behavioral characteristics of vasopressin-deficient rats (Brattleboro strain). AB - A group of vasopressin-deficient rats (Brattleboro strain--DI) and a group of normal Long-Evans rats were successively evaluated on visual discrimination, olfactory discrimination, delayed alternation at short and long intertrial intervals (ITIs), approach-avoidance conflict in a straight runway, and open field behavior. It was found that DI rats adapted more slowly than normal rats in the T-maze, in the straight runway, and they were slower to emerge into the open field. The DI rats were impaired relative to normal animals on the discrimination tasks (visual and olfactory), but they were not impaired on delayed alternation (at least for short ITIs). DI rats also showed better retention of the punishment effect in the approach-avoidance conflict test than normal animals. It is suggested that DI rats have defective reference-memory mechanisms, fairly intact working-memory processes and altered adaptability (timidity or cautiousness). PMID- 7470952 TI - Redefinition of the location of the dorsal (motor) nucleus of the vagus in the rat. AB - The rostrocaudal extent of the preganglionic parasympathetics of the vagus nerve (DNV, the dorsal nucleus of the vagus, also called the dorsal motor nucleus of the vagus) was traced using the retrograde horseradish peroxidase (HRP) transport method. Labeled cells were found within, and both rostral and caudal to, the classically-defined DNV. Cell bodies of origin of the preganglionic parasympathetics of the vagus formed a continuous longitudinal, spindle-shaped column extending from an area rostral to the rostralmost border of the inferior olivary nucleus caudally through the nucleus commissuralis (NC) of the lower medulla, and continuing in an attenuated fashion in the dorsal commissural gray (CG) of the cervical spinal cord. The rostral quarter of this cell column consisted of only a few cells lateral to the fourth ventricle in the mid-medulla. The middle one-half of the DNV cell column included the classically-defined dorsal nucleus of the vagus, located dorsolateral to the hypoglossal nucleus and lateral to the fourth ventricle rostrally, and located dorsolateral to the hypoglossal nucleus and the central canal caudally. In the caudal medulla, the caudal quarter of the cell column consisted of cell bodies in the NC. Scattered labeled cell bodies were found in the CG of the spinal cord, extending caudally to the C3--C6 level. PMID- 7470953 TI - A simple method for glass insulating tungsten microelectrodes. AB - A method for insulating tungsten wires with glass by collapsing a glass capillary tube around the wire is described. A common laboratory electrode puller can be used. A new method for removing the glass from the tip using hydrofluoric acid is also described. PMID- 7470954 TI - Olfactory receptor response characteristics: a reevaluation. AB - The activity of single olfactory receptor cells previously reported was reexamined using Guttman-Lingoes multidimensional scaling, cluster analysis and factor analysis. These analyses were compared and contrasted with similar analyses using another olfactory unit data set and a color vision receptor data set. Questions were raised regarding the interpretation of the results of these analyses for any of the data sets employed. PMID- 7470955 TI - Effects of medial preoptic lesions on male mouse ultrasonic vocalizations and copulatory behavior. AB - In male mice, lesions in the medial preoptic area (MPA) reduced the proportion of animals showing mounting behavior but failed to abolish ano-genital exploration of the female. The lesions did not effect the amount of 70 kHz ultrasound elicited by intact females or by soiled bedding obtained from female-occupied cages. These results suggest components of male sexual behavior are mediated by neural systems outside the MPA. PMID- 7470958 TI - Hypertension induced by electrical stimulation of the subfornical organ (SFO). AB - In urethane anaesthetized rats, electrical, low intensity stimulations using bipolar concentric electrodes or monopolar glass pipettes were applied to SFO and to surrounding structure's populations. Blood pressure, recorded from the femoral artery, was systematically and rapidly increased when the electrical, as well as the mechanical (electrode displacements) stimulations were located within the SFO populations. Remote from the SFO, stimulations either were ineffective or produced non reliable responses. SFO cells seen to be involved in the defense against all hydroionic deficiencies, including those of blood pressure. PMID- 7470957 TI - Circadian rhythms in vasopressin deficient rats. AB - Circadian rhythms in wheel running and drinking behavior were investigated using heterozygous an homozygous (diabetes insipidus) female Brattleboro rats Despite the lack of vasopressin in the suprachiasmatic nuclei of the diabetic rats, they showed coherent rhythms, both in cyclic light and in constant light. However, the periods of the free-running rhythms were longer for the diabetic rats, they were less active, and, of course, were severely polydipsic. Replacement treatment with systemic infusions of vasopressin reversed the polydipsia but did not affect the other measures. PMID- 7470956 TI - Distal infrapyramidal granule cell axons possess typical mossy fiber morphology. AB - Using both the Timm's sulfide silver and anterograde horseradish peroxidase (HRP) histochemical techniques, we have demonstrated that granule cell in the dentate gyrus of the rat have topographical projections to both suprapyramidal and distal infrapyramidal terminal fields in the rostral one-third of the hippocampus. An anterograde HRP technique modified for visualization of axonal morphology indicated that intrapyramidal mossy fiber axons possess classic periodic swellings characteristic of suprapyramidal mossy fibers. Most distal infrapyramidal mossy fiber axons appear to be in position to make synaptic contact with deeper-lying neurons in the pyramidal cell layer. PMID- 7470959 TI - Comparative effects of amantadine and amfonelic acid on dopamine metabolism in rat brain. AB - Amantadine (50 and 100 mg/kg IP) did not change basal levels of DOPAC (3,4 dihydroxyphenylacetic acid) or HVA (homovanillic acid) in rat brain and did not alter the increase in DOPAC levels resulting from dopamine receptor blockade by spiperone. The increase in HVA levels produced by spiperone was significantly, but only slightly, enhanced by amantadine at these doses. In contrast, amfonelic acid at a dose of 5 mg/kg increased DOPAC and HVA levels and enhanced several fold the spiperone-induced elevation of DOPAC and HVA levels. Amfonelic acid at at 2.5 mg/kg dose enhanced the turnover of dopamine (measured by the decline in dopamine concentration after inhibition of tyrosine hydroxylation with alpha methylytyrosine) in spiperone-treated rats, whereas amantadine (100 mg/kg) had a smaller and not statistically significant effect. Neither compound affected significantly dopamine levels in a alpha-methyltyrosine-treated rats not given spiperone. Although amantadine has been observed by earlier workers to enhance the stimulated release of dopamine, it does not appear to act in a manner similar too amfonelic acid. PMID- 7470960 TI - The influence of unilateral 6-OH-dopamine lesions of the substantia nigra in the absence of the telencephalon. AB - Rats were injected unilaterally into the rostral substantia nigra pars compacta with 8 micron of 6-OHDA. Those animals that conformed to be behavioral criteria for an effective nigrostriatal dopamine lesion by turning ipsilaterally to the injected side upon IP amphetamine, and contralaterally upon apomorphine injection were bilaterally detelencephalized (neocrotex, hippocampus, striatum, septum, amygdaloid complex were removed). Most detelencephalized animals exhibited spontaneous turning to the contralateral side, i.e. no longer behaved like 6-OHDA lesioned rats, but instead, acted like animals with nigral kainic acid or electrolytic lesions. Amphetamine (2-3 mg/kg) increased general activity but no longer influenced turning. Apomorphine in doses of 2-3 mg/kg completely suppressed all motility. In small doses (0.1-0.2 mg/kg) it reversed the spontaneous contraversive turning. This effect could be blocked by haloperidol (0.1 mg/kg) pretreatment. High doses of haloperidol (5.0-7.5 mg/kg) reversed the direction of circling from contraversive to ipsiversive. Arecoline (10-12 mg/kg) induced tremor as in normal animals. Atropine (50-100 mg/kg) did not affect turning, but increased activity level in the thalamic rats. PMID- 7470961 TI - The brain and subarachnoid space as possible sites for endocrine tissue transplantation. AB - Immunological rejection has been the major problem limiting the successful transplantation of tissue from one animal to another. Recent technological developments, combined with the use of the central nervous system as an immunologically privileged site, suggest that it might be possible to achieve long-term survival of hormone-secreting tissues, between two gentically dissimilar animals, if these tissues are transplanted to the brain and subarachnoid space of the host. The physiological parameters that should be considered in the clinical application of a transplant of this type are discussed. PMID- 7470963 TI - An apparatus for observing behavior of pouch-young opossums. AB - Opossum fetuses undergo most of their neurological development within the mother's pouch. This extrauterine growth offers unique opportunities not shared by placenta mammals for early behavior studies. Described here are an apparatus and technique for observing the behavior of pouch young. The apparatus restrains the mother and retracts the pouch flaps within an environmentally controlled setting similar to that inside the mother's pouch, allowing the opossum young to be observed in an almost natural state. The investigator has ready access to the young pouch through side windows that have slotted curtains to prevent sudden changes in the chamber environment. Because this apparatus provides a simple noninvasive approach, fetal behavior can be observed long-term, encompassing the entire sequence of behavior development within a chosen time-interval. PMID- 7470962 TI - An improved technique for brain microdissection and subsequent histological control. AB - A microdissection technique is described which allows removal of discrete brain regions in the cryostat under stereotaxic control. In addition, thin slices (20 micron) from the upper and lower surfaces of the dissected tissue can be preserved for subsequent staining and assessment of possible contamination by surrounding tissue. PMID- 7470964 TI - [Surgical management of inborn stenosis of the aortic valves (author's transl)]. PMID- 7470965 TI - [Transpubic approach to the membraneous urethra in posttraumatic stricture (author's transl)]. PMID- 7470966 TI - [Surgical experiences with the sternum (author's transl)]. PMID- 7470967 TI - [Concerning the problem of massive bleeding from gastroduodenal ulcers (author's transl)]. PMID- 7470968 TI - [Further observations on metabolic changes in the peroperative period (author's transl)]. PMID- 7470969 TI - [When is Rodney Smith's operation for reconstruction of the biliary ducts indicated (author's transl)]. PMID- 7470970 TI - [Concerning the problem of intestinal necroses (author's transl)]. PMID- 7470972 TI - [Malignant hyperthermia syndrome (author's transl)]. PMID- 7470971 TI - [Experiences with profundoplastics in treating ischemic diseases of the lower extremities (author's transl)]. PMID- 7470973 TI - [Contribution to indications of proximal selective vagotomy (author's transl)]. PMID- 7470975 TI - [Poland's syndrome in our population (author's transl)]. PMID- 7470974 TI - [Primary spinocellular carcinoma of the skin on the finger (author's transl)]. PMID- 7470976 TI - Effect of neonatally administered testosterone propionate on immunological response in white leghorn chickens. (Gallus domesticus). PMID- 7470977 TI - Non-histone chromatin proteins (NHP) of mice thymocytes after in vivo immunization and during in vitro proliferation by PHA. PMID- 7470978 TI - [The work framework of the French physician]. PMID- 7470979 TI - [The work load of the medical corps]. PMID- 7470980 TI - [The reading of medical publications--continuing medical education group medicine -auxiliary personnel]. PMID- 7470981 TI - [Methodologic presentation and representative examination]. PMID- 7470982 TI - Premarket review of intraocular lenses. PMID- 7470983 TI - Consecutive exotropia after correction of hyperopia. AB - A study was done of 22 children in whom exotropia developed following correction of hyperopia because of accommodative esotropia that had not been corrected surgically. The early onset of esotropia and high degree of hyperopia seemed to predispose to the spontaneous development of exotropia. Reduction of the hyperopia by 50% to 60% resulted in satisfactory realignment of the eyes and some form of binocularity. PMID- 7470984 TI - Visual results after removal of congenital cataracts before the age of 1 year. AB - Six patients underwent removal of congenital cataracts (Unilateral in three and bilateral in three) from otherwise normal eyes in the first year of life. Good corrected vision, including 6/9 in one of the unilateral cases, was obtained in each aphakic eye. A more aggressive approach to eliminating the sensory obstacles to seeing should include the removal of dressings 1 day after the operation, the use of soft, permanent-wear contact lenses within a few days thereafter and reduction of the time between the operations on bilateral cataracts to 5 days. In unilateral cases an occlusive patch must always be worn over the normal eye 90% of the waking day, even during illnesses. PMID- 7470985 TI - Chemiluminescent evaluation of the human lacrimal outflow system. AB - A chemiluminescent material, Cyalume, has been used to demonstrate the structure and outflow patency of the human lacrimal drainage system. In this paper the details of this method are outlined and some of the possible benefits and limitations are discussed. Work is in progress to improve the techniques and to evaluate the toxicity of the chemiluminescent material in animals. It is hoped that these studies will lead to the development of a simple and rapid office procedure for assessing the structure and patency of the lacrimal tracts and simultaneously evaluating any obstruction. PMID- 7470986 TI - Modelling cortical cataractogenesis: 2. In vitro effects on the lens of agents preventing glucose- and sorbitol-induced cataracts. PMID- 7470987 TI - [Transitory closed-angle glaucoma following a central retinal vein thrombosis]. AB - This article reports a case in which an occlusion of the central retinal vein of one eye was accompanied by transitory shallowness of the anterior chamber, partial closure of the angle and elevation of the intraocular pressure. PMID- 7470988 TI - Factors influencing the release of labelled gamma-aminobutyric acid and acetylcholine evoked by electrical stimulation with alternating polarity from rat cortical slices. PMID- 7470989 TI - Species differences of amobarbital metabolism: dihydroxyamobarbital formation. AB - Amobarbital is almost completely metabolized in animals and man. The major metabolite is 3'-hydroxyamobarbital. The second major metabolite in dog, mice, hamsters, guinea pigs, and rats could be identified as 3',4' dihydroxyamobarbital. A striking contrast between man and animals is the virtual absence of the diol in man and absence of amobarbital-N-glucoside in animals. PMID- 7470990 TI - A comparison of the dipsogenic responses of male and female rats to a variety of stimuli. AB - Female rats subjected to 48 h of water deprivation drank more than similarly deprived males. There was no sexual difference in the dipsogenic responses to intracellular dehydration produced by intravenous hypertonic NaCl but females drank considerably more than males in response to the extracellular stimuli of hyperoncotic polyethylene glycol and angiotensin II. It is concluded that intact adult female rats are dipsogenically more responsive than adult male rats to stimuli acting through the pathways of extracellularly induced thirst but not to those arising from the intracellular fluid compartment. PMID- 7470991 TI - Effect of heavy metals and lanthanum on sugar transport in isolated guinea pig left atria. AB - The effect of heavy metals on sugar transport in fully resting guinea pig left atria was studied by measuring the tissue-medium distribution of 3-methylglucose. Basal sugar transport was increased significantly by all heavy metals tested (Co2+, Ni2+, Zn2+, Mn2+ (2 mM) and by La3+ (0.05 mM) but 1 mM La3+ had no effect. The stimulation of sugar transport by insulin, hyperosmolarity, K+-free medium, or 10(-5) M ouabain was strongly antagonized by Ni2+, Zn2+, and La3+ but was unaffected by Co2+ and Mn2+. The heavy metals did not affect intracellular Na2+ and K+, whether in the basal state or when the Na+ pump was depressed by ouabain or K+-free medium. The data suggest that Ca2+ antagonistic ions may affect sugar transport both by inhibiting Ca2+ influx and by competing with Ca2+ for specific binding sites presumably involved in the regulation of sugar transport. PMID- 7470992 TI - Sensory-motor processing in the caudate nucleus and globus pallidus: a single unit study in behaving primates. AB - Monkeys were prepared for chronic recording of single neurons in the caudate nucleus (Cd) or globus pallidus (GP) during learned wrist flexion-extension movements triggered by visual and somatic sensory inputs. Almost two-thirds of GP cells and more than one-third of Cd cells modified their discharge during these tasks. Three categories of response types were observed. The first was movement related. The second type was event related, in which the cells responded to either the onset or offset of the sensory inputs regardless of the correcting movement direction. A third type combined elements of the first two categories and was termed complex. These cells responded to complex abstractions of the sensory-motor event. A latency analysis indicated that the majority of cells was not involved in initiating movements but may have participated in movement execution. The results of this experiment suggest that during voluntary movement the basal ganglia activity is correlated with motor outputs, sensory inputs, and perceptual abstractions of these sensory-motor events. As such the results are compatible with an influence by diverse regions of cerebral cortex on basal ganglia neurons during the movement control process. PMID- 7470993 TI - Effects of rate of blood flow on fractional extraction and on uptake of infused noradrenaline by brown adipose tissue in vivo. AB - The rate of blood flow (Q) to interscapular brown adipose tissue (IBAT) and the arteriovenous difference in plasma noradrenaline (NA) across the tissue were measured in warm-acclimated (WA) or cold-acclimated (CA) rats during infusion of NA at doses of 1-12.5 ng min-1 g-0.74 (approximately 0.2-2.7 micrograms min-1 kg 1) and in the period of steady calorigenic response associated with steady concentration of NA in arterial plasma (ANA). ANA was linearly related to the dose of NA. Calorigenic response, percentage of cardiac output to IBAT, and Q per gram of IBAT were sigmoid functions of ANA and at their maxima were about 2.5 times greater in CA than in WA rats. The rate of uptake of NA by IBAT increased with ANA and Q, each of which had a major influence on rate, but the coefficient of extraction of NA by the tissue (ENAIBAT) declined. Measurements in rats given a dose of propranolol that partially inhibited the NA-induced increase in Q to IBAT indicated that the decline in ENAIBAT was attributable primarily to the increase in Q rather than to increasing saturation of uptake mechanisms. Diffusion-limited extraction of NA is the probable basis for the effect of Q on ENAIBAT. Possible implications of flow-dependent extraction of NA in studies involving measurements of the uptake of exogenous NA by tissues or organs are discussed. PMID- 7470994 TI - Steroid anaesthetics: inhibition of depolarization-secretion coupling at the mouse motor nerve terminal. AB - The coupling between nerve terminal depolarization and quantal secretion of acetylcholine at the mouse neuromuscular junction was estimated by measuring the multiplication of the frequency of miniature end-plate potentials (m.e.p.p.s) produced by increasing the concentration of calcium in the medium from 0.1 to 1.0 mM in the presence of 15 mM potassium. Depolarization-secretion coupling was inhibited by the anaesthetic steroids progesterone, pregnanedione, and alphaxalone. The nonanaesthetic steroid delta 16-alphaxalone also inhibited depolarization-secretion coupling with the same potency as alphaxalone. This results indicates that inhibition of depolarization-secretion coupling in nerve terminals is unlikely to play a major role in the production of anaesthesia. PMID- 7470996 TI - Erythrocyte life-span in dystrophic hamsters. AB - [3H]Diisopropylfluorophoshate was used to label erythrocytes from normal and dystrophic hamsters. The mean erythrocyte life-span in dystrophic animals was 57.3 +/- 0.7 days whereas in normal animals it was 50.1 +/- 1.6 days. PMID- 7470995 TI - Substrate repletion in rat myocardium, liver, and skeletal muscles after exercise. AB - Carbohydrate and lipid substrates were measured in rats during recovery following exercise or a 24-h fast and compared with values from time-matched control (rested, fed) rats. After exercise muscle glycogen recovered at the expense of liver glycogen repletion. Myocardial glycogen supercompensated whereas soleus, red vastus lateralis (RVL) and white vastus lateralis glycogen merely returned to control levels. A similar recovery pattern occurred after fasting with refeeding promoting glycogen synthesis in the liver, skeletal muscles, and even in the myocardium, where glycogen had already been elevated by the fast. Both soleus and RVL muscles, along with the myocardium, exhibited glycogen supercompensation. Both exercise and fasting increased plasma free fatty acid (FFA) levels which favor myocardial glycogen synthesis. Unchanged tissue triglycerides and relatively stable blood glucose levels suggest that these are unlikely influences on glycogen recovery. It is concluded that exercise per se is unlikely to induce glycogen supercompensation in skeletal muscles though myocardial glycogen supercompensation readily occurs, that food restriction prior to exercise quantitatively affects substrate recovery though its impact could go unnoticed because of the qualitative similarities between substrate recovery following exercise or fasting, and that FFA is the only major energy substrate concurrently changing with glycogen after exercise or fasting which could facilitate glycogen synthesis. PMID- 7470997 TI - Learning preferences and continuing medical education. PMID- 7470998 TI - "Doctor, my feet hurt". PMID- 7470999 TI - Dying and the family physician. PMID- 7471000 TI - How to read clinical journals: I. why to read them and how to start reading them critically. PMID- 7471003 TI - Skull roentgenography for patients with head trauma: the use of high-yield criteria. PMID- 7471001 TI - Continuing medical education in Maritime Canada: the methods physicians use, would prefer and find most effective. AB - A study of the continuing medical education of practising physicians in Nova Scotia, New Brunswick and Prince Edward Island was conducted in 1979-80 by means of a mailed questionnaire. Most of the responding physicians ranked reading as the method most used to update knowledge (73.3%) and skills (55.7%); courses and informal instruction were in second place for updating knowledge and skills respectively, ranked most used by 9.3% and 17.1%. With unlimited time and funds 38.0% and 20.5% of the physicians would still most prefer to read to update knowledge and skills respectively. However, 35.2% would most prefer to attend courses to update knowledge and 26.9% and 24.8% would most prefer to do clinical traineeships or attend courses to update skills. When asked what method of learning had provided the most impetus to change their ways of managing patients, 42.5% chose reading, 18.8% courses, 14.6% informal discussions and 12.4% formal consultations. Appropriate developments would therefore include improving methods of providing physicians with the best information available when it is needed, removing roadblocks to participation in continuing education programs, matching individual learning styles to programs of learning, training physicians as peer tutors and helping consultants become better instructors through written consultations. PMID- 7471002 TI - Left ventricular function during graded exercise in patients with coronary artery disease and in control subjects. AB - The left ventricular function of 30 patients with coronary artery disease and 11 control subjects was studied by electrocardiography gated cardiac blood pool scintigraphy as the participants lay on their backs and either rested or exercised on a cycle ergometer at graded levels on intensity. The control subject showed a progressive increase in ejection fraction from rest (51% +/- 7%) to intermediate (56% +/- 10%, P less than 0.05) and maximum levels of exercise (64% +/- 10%, P less than 0.001). All the patients showed a decrease in ejection fraction from rest (42% +/- 16%) to their maximal level of exercise (36% +/- 11%, P less than 0.001). However, the response of some of the patients to intermediate exercise ranged from a decrease or no change to an increase in ejection fraction. Thus, exercise at maximal intensity is necessary to induce the left ventricular dysfunction that is diagnostic of coronary artery disease. PMID- 7471004 TI - A complication of catheterization of the left internal jugular vein. PMID- 7471005 TI - The changing role of the Canadian physician in the Third World. PMID- 7471006 TI - Foreign medical graduates: screening out the poorly trained. PMID- 7471007 TI - Preparing a testamentary letter for your family. PMID- 7471008 TI - Learn to run your office like a business. PMID- 7471009 TI - Difficulties in commitment of the mentally ill. PMID- 7471010 TI - Author, editor and reviewer: how manuscripts become journal articles. PMID- 7471011 TI - The periodic health examination. PMID- 7471012 TI - Inadequacy of current requirements for licensure in family medicine. PMID- 7471013 TI - The seeing-eye dog: an infection risk in hospital? AB - Because an increasing number of blind people are using guide dogs, hospital administrators may receive more frequent requests to allow a guide dog to accompany its blind owner during visit with a hospital patient. This article considers the risks and problems for all concerned if seeing-eye dogs are permitted in hospitals, and proposes a reasonable set of guidelines for hospital personnel to apply when considering these requests. PMID- 7471014 TI - How to read clinical journals: II. To learn about a diagnostic test. PMID- 7471015 TI - Involuntary commitment in Ontario: some barriers to the provision of proper care. AB - The Ontario Mental Health Act, as amended in November 1978, provides strictly defined criteria for involuntary commitment for psychiatric assessment; the assessment can last up to 5 days. These criteria fail to cover a number of patients who are, in the author's opinion, in need of compulsory assessment or treatment. Four cases in which there was serious difficulty in giving proper care are described in this paper. The difficulties are discussed and improvements in the criteria for involuntary assessment recommended. PMID- 7471016 TI - Compliance by physicians with the 1978 Ontario Mental Health Act. AB - In a survey of 102 patients admitted involuntarily to a psychiatric hospital, a statement of potential danger, required by law, was found to be provided on all the committal forms. Closer enquiry suggested that although 89% of these patients were admitted involuntarily in accordance with strict legal requirements the legal justification for involuntary admission was questionable in 11 cases. Of these 11 admissions, 10 were thought to be warranted if medical considerations were given priority and on humane grounds. For 6 of the 10 patients treatment was clearly beneficial. One patient suffered hardships as a result of his involuntary admission that were not compensated for by any likelihood that he could have benefited medically from psychiatric care. Apparently humane considerations and the possibility that a mentally ill person could benefit from treatment led to some committals that were legally uncertain. It is concluded that the Mental Health Act should be revised to take account of such situations. PMID- 7471017 TI - Oxalate nephropathy due to gastrointestinal disorders. AB - Renal failure secondary to oxalate interstitial nephritis developed in three patients with malabsorption and steatorrhea following a jejunoileal bypass, extensive small intestine resection and a partial gastrectomy. Hyperoxaluria was documented in two of the cases. The possibility that this complication can occur in patients after a jejunoileal bypass operation is now recognized. This report shows that it can also occur in patients with other bowel disorders that cause malabsorption and steatorrhea. Since the prognosis for patients with oxalate nephropathy is poor, renal function should be closely monitored in patients who are at risk because of these disorders. Therapy should be directed at correcting malabsorption, steatorrhea and hyperoxaluria. When the renal function of patients with a jejunoileal bypass continues to decline despite intensive medical therapy, restoration of bowel continuity is strongly recommended. PMID- 7471018 TI - Hyperglycemia in anticholinesterase poisoning. PMID- 7471019 TI - Miliary tuberculosis presenting as adrenal failure. PMID- 7471020 TI - Investor-owned hospitals: efficient management means profit. PMID- 7471021 TI - The pharmaceutical industry: promoting research in the '80s. PMID- 7471023 TI - Telemedicine provides new dimensions in CME in Newfoundland and Labrador. PMID- 7471022 TI - Why abandon rotating internship programs? PMID- 7471024 TI - The first recorded case of osteitis fibrosa cystica. PMID- 7471025 TI - Concepts and practice of prognosis in psychiatry. AB - The frequency with which prognosis is made in psychiatry is not known. This paper reports a study of the frequency and content of prognosis as recorded by staff psychiatrists and residents in a psychiatric teaching hospital. In addition, a review was carried out on the use of prognosis in ten psychiatric textbooks. The findings suggest that closer attention should be given to the teaching of prognosis. There is a clear need to consider moving away from the close dependence of prognosis on diagnosis, and from the unidimensional view of prognosis. Reasons are given for developing a prognosis, and suggestions are made regarding the organization of the prognostic statement. PMID- 7471026 TI - Treatment of the elderly on a general hospital psychiatric short-stay unit. AB - Two out of seven general hospital psychiatric units and the local mental hospital in the Vancouver area set an arbitrary age limit of 65 or 70 on admissions. Administrative reasons given for this include the presumed likelihood of organicity and chronicity, implying difficulties in disposition, and problems in maintaining an active therapeutic milieu with patients of other ages. Data are presented on 40 patients who were over the age of 65 and accounted for 8% of all admissions during 1977 to a Psychiatric Short-Stay Unit in a downtown general hospital. It was found that over three-quarters of the admissions had a primary functional diagnosis, that the mean duration of stay was 5.73 days, that two thirds of the patients were able to return to their former place of living on discharge and that over one-half of the patients located on follow-up, 18 to 24 months later, were still residing in the same situation. During hospitalization, almost one-half of the patients received no psychotropic prescription and almost one-third, no physical prescription. Some of the psychosocial factors in the treatment program are presented. It is concluded that this preliminary study suggests that the administrative apprehensions in relation to treatment of the elderly in general hospital psychiatric units and mental hospitals can be countered. PMID- 7471027 TI - Hemolytic anemia associated with chlorpromazine therapy. AB - A rare case of hemolytic anemia is presented which occurred on exposure to chlorpromazine and resolved when the medication was discontinued. Another complication, absent gag reflex, is also discussed. PMID- 7471028 TI - Women and depression. PMID- 7471029 TI - Toward integrating psyche and soma: psychoanalysis and neurobiology. AB - The brain is the "key organ" for understanding mind/body/illness relationships. During the past two decades neurobiological research has generated a plethora of new data and concepts which have increased tremendously our knowledge of the functioning brain. As a result the psychoanalytic view of the relationship between mind and brain may seem at risk of becoming outmoded. Yet while psychoanalytic theory may no longer be wholly tenable, psychoanalysis continues to offer interesting and heuristically valuable isomorphic models of cortical function. On the other hand neurobiology provides a corrective influence on psychoanalytic concept-building, causing theory to be refined as it is tested against the results of research. One possible result of interdisciplinary cross fertilization is that a revised theory of the function of dreams and fantasy may throw light on the vicissitudes of somatic experience, and the pathogenesis of psychophysiological disorder. PMID- 7471030 TI - Women and depression. PMID- 7471031 TI - Canadian psychiatry of the 80's: gateway or goal. PMID- 7471032 TI - The early phase of psychotherapy with antisocial early adolescents. AB - Adolescence is a developmental stage with specific defenses, styles, and ego functions and thus requires a specific psychotherapeutic technique. The task of early adolescence is the establishment of a sense of autonomy during major biological, psychological, familial and social changes. Antisocial behaviour is understood as having multiple etiology and to be a manifestation of different diagnostic entities including reactive disorder, depression, neurosis, narcissistic, impulsive and borderline personalities, or psychosis. The main goal of the early phase of therapy is the establishment of a therapeutic relationship. This is achieved through the therapist's ability to help the adolescent to talk and explore, without fear of exposure, in the context of an active, empathic relationship. Specific guidelines helpful in establishing this relationship in the early phase of adolescent therapy are discussed. PMID- 7471033 TI - Neuropsychiatric and electroencephalographic aspects in the diagnosis of complex partial seizures. AB - The authors review from a psychiatric perspective some of the electroencephalographic techniques that may assist the clinician in diagnosing complex partial seizures (temporal lobe epilepsy). Routine scalp electroencephalograms may not detect temporal lobe EEG abnormalities, leading to the false impression of pseudo or hysterical seizures. Knowledge of the advantages of a sleep recording, specialized EEG electrodes (nasopharyngeal, sphenoidal), and natural or pharmacological activation techniques will increase one's diagnostic acumen. The authors discuss the appropriate use of these methods and their relevance to psychiatry. PMID- 7471034 TI - Training in large group therapy. AB - The widespread use of large group meetings as a treatment modality has exceeded the availability of staff trained in large group methods. The basic elements of an inservice training program are regular attendance by experienced senior staff who serve as role models, the serious and open use of review meetings following each large group meeting, and the establishment of group norms that facilitate functioning. A few crucial norms are identified and their importance is illustrated with examples from large group meetings. PMID- 7471035 TI - LSD and onset of schizophrenia. AB - In a matched controlled study 37 chronic schizophrenics who had taken LSD preceding the onset of their illness were compared with schizophrenics who had not. There were no significant differences between the groups either on age at onset of symptoms or for age at first admission. PMID- 7471036 TI - Physical disorders among emergency psychiatric patients. AB - An examination of patients with physical disorders requiring immediate treatment or investigation was undertaken in the Psychiatric Emergency Service of the Royal Ottawa Hospital and found to comprise 7% of the patient population, or one visit per day. Those with physical problems differed from the rest of the emergency patients in that a greater number were over age 60 and were diagnosed as alcoholic or chronic organic brain syndrome. Socio-demographic characteristics were examined as well. The presenting problem was accurate regarding a physical basis in only two-thirds and the disorder could, in some way, be related to a psychiatric diagnosis in 75% of the patients. Emphasis is placed on the fairly high frequency of immediate physical attention required by these patients who come to the psychiatrist. Consequently, psychiatric assessment and management is not the priority for this group and other skills of the psychiatrist are necessary in treating the patient. PMID- 7471037 TI - Neurosyphilis in a young man. AB - New cases of neurosyphilis are comparatively rare. The epidemiology and clinical presentation of neurosyphilis are described. The article describes the presentation of general paresis in a 34 year old man, 10 years after he had been ineffectively treated with penicillin for secondary syphilis. The various laboratory tests for syphilis are reviewed. The Venereal Disease Research Laboratories (VDRL) is currently the best screening test and the Fluorescent Treponema Antibody-Absorption test (FTA-ABS) is the current recommended specific test for syphilis. Central nervous system involvement is confirmed in the laboratory by a positive VDRL on the cerebrospinal fluid. Physicians should be on the alert for this cause of dementia which may, once again, be increasing in frequency. PMID- 7471038 TI - A case of lyssophobia. PMID- 7471039 TI - Multiple personality. PMID- 7471040 TI - Mesothelioma associated with the shipbuilding industry in coastal Virginia. AB - A case-control study was undertaken to clarify reasons for a four-fold increased incidence of mesothelioma discovered among white males in coastal Tidewater, Va., from 1972 to 1978. Sixty-one cases were identified. Interviews with next of kin revealed that the excess was linked to employment in area shipyards. Three fourths of the cases had been employed in the shipbuilding industry, nearly all beginning employment prior to 1950. Most were career employees, but an increased risk was also found among those who worked only temporarily, mainly during World War II, and were reportedly exposed to asbestos. More of the cases than controls were pipecoverers or pipefitters, but cases were reported to work in a variety of shipyard trades. Few of the mesothelioma cases were heavy smokers, a trend that may be related in part to the competing risks for fatal diseases caused by the interactions of smoking and asbestos exposure. Information obtained by interview for five of the six white females diagnosed with mesothelioma revealed that the husband of four had been employed in the shipbuilding industry. PMID- 7471041 TI - Structural and functional effects of adriamycin on cardiac cells in vitro. AB - The effects of Adriamycin (ADR) on the heart, seen clinically as transient electrocardiographic changes and cardiomyopathy, have been simulated in an in vitro cardiac cell system. Structural and functional alterations in cultured heart cells can be dissociated based upon ADR dose and length of exposure. At high ADR doses (100 to 200 micrograms/ml), cessation of beating was rapid, and structural changes consistent with the in vivo cardiomyopathic picture (vacuolization and nucleolar fragmentation) were observed. At low ADR doses (0.1 to 0.5 micrograms/ml), arrhythmias were produced in the absence of ultrastructural changes (within 48 hr); the incidence and severity of the arrhythmias were demonstrated to be dose dependent. Continued treatment of cultures at low dose levels for sustained periods of time (up to 17 days) resulted in a striking loss of muscle fiber without concomitant vacuolization and nucleolar fragmentation. An intermediate ADR dose of 10 micrograms/ml for 1 hr exposure caused vacuolization and cessation of beating, with lysis of cells within 72 hr. The parallel between the effects of ADR on in vitro cardiac cell structure and function with those seen in vivo suggests that this simple system may have value in studies directed towards the mechanism of ADR-induced cardiac toxicity and in the screening of anthracycline analogs for their potential effects on the heart. PMID- 7471042 TI - Exceptionally high tumor-initiating activity of benzo(c)phenanthrene bay-region diol-epoxides on mouse skin. PMID- 7471043 TI - Increased cytidine 5'-triphosphate synthetase activity in rat and human tumors. PMID- 7471044 TI - Macrophage functions in beige (Chediak-Higashi syndrome) mice. AB - Beige (C57BL/6J-bgj/bgj) mice are the murine counterpart of the Chediak-Higashi syndrome, exhibiting abnormal lysosomes in phagocytes. These mice, however, responded normally to Corynebacterium parvum killed bacterial vaccine with splenomegaly and an increase in peritoneal macrophages. The C. parvum-elicited macrophages showed normal immunoglobulin G Fc- and C3b-mediated rosettes and phagocytosis. The antitumor action of the macrophages was assessed against the Lewis lung carcinoma, a tumor that is sensitive to inhibition by activated macrophages both in vitro and in vivo. The elicited macrophages from beige mice showed delays in vitro in cytostatic and cytotoxic activity against the tumor cells, as compared with C57BL/6J-+/+ control mice. The early delays in activity disappeared after 12 to 24 hr, when antitumor activity was similar to that exhibited by C. parvum-elicited macrophages of the C57BL/6J-+/+ mice. These delays in antitumor activity of beige mouse macrophages may be analogous to the delays observed in the bactericidal activity of beige mouse granulocytes. The ultimate antitumor activity, however, was comparable in beige and +/+ C. parvum elicited macrophage. Moreover, the resistance of mice in vivo to the Lewis lung tumor was not markedly impaired. The growth of the primary tumors and the mean times to death of the tumor-bearing animals of both strains were similar. PMID- 7471045 TI - Reduced capacity to repair irradiated adenovirus in fibroblasts from xeroderma pigmentosum heterozygotes. AB - Xeroderma pigmentosum (XP) is one of a number of autosomal recessive syndromes in humans characterized by a marked predisposition to cancer. Fibroblasts from these patients show a defect in DNA repair. The XP heterozygotes also show elevated skin cancer incidence, but reports concerning their DNA repair capacity are conflicting. In this study, the DNA repair capacity of four XP heterozygotes was examined using a sensitive host cell reactivation technique. Unirradiated and irradiated suspensions of adenovirus type 2 (Ad 2) were assayed for their ability to form viral structural antigens in fibroblasts from XP heterozygotes, XP homozygotes, and normals. A reduced host cell reactivation (of viral structural antigen production) for both ultraviolet- and gamma-irradiated Ad 2 was detected in four XP heterozygotes representing three different complementation groups as well as their XP homozygous children. The doses necessary to reduce the survival of viral structural antigen production by irradiated AD 2 to 37% in the XP heterozygous strains were expressed as a percentage of that obtained in normal strains and ranged from 55 to 82% for ultraviolet-irradiated Ad 2 and 71 to 79% for gamma-irradiated Ad 2. These results add further support to a direct relationship between cancer proneness and DNA repair defects and show the merits of this host cell reactivation technique in identifying XP heterozygotes. Identification of XP heterozygotes is of considerable public health interest not only in genetic counseling but also in the prevention of cancer. PMID- 7471046 TI - Selective dissociation and characterization of cells from different regions of multicell tumor spheroids. AB - A technique has been developed which takes advantage of the spherical symmetry of EMT6/Ro multicellular tumor spheroids to isolate subpopulations of cells originating from various locations in the spheroid structure. The method involves gently exposing spheroids to a dilute trypsin solution at 18 to 20 degrees in specially designed dishes on a rotary shaker. Under these conditions, cells are released only from the outer spheroid surface; successive trypsin treatments dissociate cells from increasing depths in the spheroid. Measurements of the spheroid diameter and rate of cell dissociation demonstrated the reproducibility of the technique. Observations of histological sections showed that trypsin was active only over a small distance into the spheroid. Characterization of the cells isolated indicated that cell volume, membrane integrity, and clonogenic capacity all decreased for cells located in inner spheroid regions. Autoradiography and DNA content analysis by flow cytometry established that outer region cells were actively proliferating while inner region cells were in a nonproliferative state. There was a significant number of cells arrested with S- and G2-phase DNA contents as well as a large number arrested with a G1 DNA content, similar to recent findings for nonproliferating cells in tumors. PMID- 7471047 TI - Synergistic interactions of various doses of diethylstilbestrol and x-irradiation on mammary neoplasia in female ACI rats. PMID- 7471048 TI - Growth interaction in vivo between tumor subpopulations derived from a single mouse mammary tumor. AB - Our laboratory has previously isolated several tumor cell populations from a single, spontaneously arising mammary tumor of a BALB/cfC3H mouse and established them in tissue culture as independent sublines. These subpopulations differ according to many criteria including growth parameters and expression of tumor associated antigens. We have tested the interaction in vivo of several of these subpopulations by injecting cell suspensions of the same or different sublines into opposite flanks of BALB/cfC3H or BALB/c mice. The growth characteristics of certain subpopulations were altered by the presence of a different subpopulation on the opposite side. In order to understand the mechanism of interaction, we chose two subpopulations (410 and 168) for further study. In BALB/cfC3H mice, the presence of line 410 tumors on one flank inhibited both 410 and 168 tumors on the other flank. Line 168 tumors did not inhibit either 410 or 168 tumors. The inhibitory effect of line 410 appeared to be immunological, since (a) it was increased by injecting line 410 several weeks before line 168, (b) it was abrogated in mice subjected to 400-rad X-irradiation 2 days prior to tumor cell injection, (c) mice could be made resistant to both line 410 and line 168 tumors by implantation followed by surgical removal of line 410 but not of line 168, and (d) resistance could be adaptively transferred with lymph node cells from line 410-sensitized mice in Winn assays. Thus, immunity to tumor-associated antigens may be one way by which cells of a heterogeneous tumor can interact. PMID- 7471049 TI - Production of human chorionic gonadotropin and its subunits by human tumors growing in nude mice. AB - The production and secretion of human chorionic gonadotropin (HCG) and its subunits by human tumors growing in nude mice have been examined. JAR choriocarcinoma cells growing in nude mice produce both free alpha subunit and complete HCG, but there is a decrease in the amount of free alpha subunit relative to complete HCG produced in vivo compared to HCG subunit production by these cells growing in culture. Cell lines that produce only free alpha subunit in culture (HeLa cervical carcinoma, ChaGo bronchogenic carcinoma, and BT-20 breast carcinoma) continue to produce primarily free alpha subunit in vivo, but a small amount of HCG-beta/HCG is detectable in the 24-hr urine collected from mice bearing HeLa or ChaGo tumors. CBT cells derived from a glioblastoma multiforme produce both alpha and HCG-beta/HCG in vivo. This represents a distinct shift from the pattern of HCG subunit production by CBT cells in culture because cultured CBT cells produce only free beta subunit and do not synthesize either free alpha or complete HCG. Thus, for human tumors growing in nude mice, there appears to be a shift toward more complete HCG production and a decrease in free subunit production as compared to the pattern observed for cultured cells. PMID- 7471050 TI - Inhibition by putrescine of the induction of epidermal ornithine decarboxylase activity and tumor promotion caused by 12-O-tetradecanoylphorbol-13-acetate. AB - The induction of epidermal ornithine decarboxylase (EC 4.1.1.17) (ODC) following topical application of the tumor-promoting agent 12-O-tetradecanoylphorbol-13 acetate (TPA) to mice can be inhibited by topical application of putrescine, the product of the enzyme. The degree of inhibition depended on both the dose and the time of putrescine application; application of 20 mumol of putrescine 2 hr after TPA treatment inhibited the induction of ODC activity by 50%. TPA-induced activity of another polyamine-biosynthetic enzyme, S-adenosyl-L-methionine decarboxylase (EC 4.1.1.50), was unaffected by application of putrescine. Among several amines tested for their ability to inhibit the induction of ODC activity, spermidine, 1,7-diaminoheptane, and spermine were the most effective, causing a 90% inhibition at the 20-mumol dose. Putrescine, when added directly to the assay medium at a 100-mumol dose level inhibited by 97% the TPA-induced ODC activity, but the amount of putrescine (20 mumol) which gave 50% inhibition of the induction of ODC activity in vivo had no effect when added to the assay system. Mixing of soluble extracts from TPA-treated mouse epidermis posttreated either with acetone or putrescine or with mouse epidermis treated with putrescine alone gave essentially additive ODC activity. Furthermore, putrescine did not elicit production of detectable ODC-antizyme activity in mouse epidermis. Putrescine inhibited the formation of mouse skin papillomas promoted with TPA. Topical application of 20 and 100 mumol of putrescine 2 hr after each application of TPA to mice initiated with 7,12-dimethylbenz[a]anthracene resulted in a 30 and 80% inhibition, respectively, of papilloma formation compared to animals receiving no putrescine. PMID- 7471051 TI - Decreased uptake of 14C-labeled dicarboxylic amino acids in rapidly growing hepatomas. AB - In contrast to the increased uptake of amino acids which has been found in many neoplastic cells, we have observed a decrease in the net uptake of [14C]aspartate and [14C]glutamate in rapidly growing hepatomas relative to rat host liver. When measured 10 min after s.c. injection, the radioactivity from 14C-labeled dicarboxylic amino acids was greater in liver than in all other tissues examined (blood, skeletal, muscle, heart, spleen, lung, and brain) except kidney, where there was an approximately 2-fold greater uptake of aspartate and 10-fold greater uptake of glutamate. Mean uptakes in the rapidly growing Morris hepatomas 7288CTC and 7777 were 19 to 26% of corresponding values for the host livers. Comparison with uptake of 3H2O indicated that these low values were not solely due to differences in circulation. Decreased uptake was not accompanied by equivalent decreases in the concentration of aspartate and glutamate in the tumors. There were small changes in the net uptake of these amino acids in the slowly growing hepatoma 7787 and no significant differences in regenerating liver and hepatoma 5123C, a tumor of intermediate growth rate. The net uptake of [14C]arginine and [14C]lysine in the hepatomas was similar to that in host livers, except for a 250% increase in uptake of [14C]lysine in hepatoma 5123C. A decreased uptake of the magnitude seen with dicarboxylic amino acids in rapidly growing hepatomas has not been observed with other amino acids. PMID- 7471052 TI - Bone marrow chromosome abnormalities in breast cancer in patients following adjuvant chemotherapy. AB - Two patients developed acute bone marrow cancer following mastectomy and institution of alkylating agents as adjuvant chemotherapy. An aneuploid condition was observed in both cases, along with involvement of chromosomes 11 and 12 in structural rearrangements. Subsequent studies of 18 patients who had or had not received such therapy showed no evidence of chromosomal aberrations. However, the long-term effect of adjuvant chemotherapy in cancer patients is still of concern until additional information becomes available. PMID- 7471053 TI - Intraperitoneal hyperthermic treatment of implanted peritoneal cancer in rats. AB - The feasibility and efficacy of treating peritoneal cancer implants by applying heat to the peritoneal surfaces were studied in inbred Buffalo A rats given i.p. injections of Morris hepatoma 5123TC tumor cells. Heat was delivered to the peritoneum by contact with a heated physiological salt solution (Normosol-R) in the peritoneal cavity. A treatment temperature of 43.3 +/- 0.3 degrees was maintained for 30 min by an immersed stainless steel coil through which hot liquid circulated. Rats implanted with 0.5 to 1.0 x 10(8) tumor cells were treated at 1 to 4 hr (Group I), 4 to 5 days (Group II), and 22 to 24 days (Group III) after tumor implantation to simulate treatment for the clinical conditions of surgically spilled cancer cells, established microscopic cancer implants, and macroscopic cancer implants, respectively. A statistically significant improvement in survival was observed in Groups I and II compared with sham treated control animals; 58% of the heat-treated animals were cured. Only a slight but statistically insignificant improvement was noted in Group III. These observations indicate that i.p. surface heat treatment of peritoneal implanted cancer is feasible and effective. PMID- 7471054 TI - Inhibition of human ovarian cancer colony formation by adriamycin and its major metabolites. AB - We have examined the in vitro sensitivity to Adriamycin of human ovarian cancer colonies cloned in soft agar. In the 26 patients tested, 3 different patterns of sensitivity to Adriamycin were observed: (a) in 75% of the previously untreated patients, there was greater than 70% reduction in colony-forming cells after exposure to Adriamycin (1.0 micrograms/ml), a level which approximates the peak plasma level after i.v. therapy; (b) in all the patients who had progressive disease while on a chemotherapy regimen without Adriamycin, a greater than 70% reduction in colony-forming cells was observed only at a concentration of 10 micrograms/ml, a level not achievable by i.v. administration; (c) in 80% of patients with progressive disease after treatment with Adriamycin as part of the primary chemotherapy regimen, a 70% reduction in tumor colony-forming cells could not be achieved even at 10 micrograms/ml. These in vitro results are in agreement with clinical observations regarding the effectiveness of Adriamycin in previously untreated patients (42% response rate) with ovarian cancer as well as its ineffectiveness (0 to 6% response rate) as a second-line therapy in relapsed patients. The results also have provided a rationale for an ongoing Phase I trial of i.p. Adriamycin in patients with ovarian cancer from Group b above since cytotoxic levels can be produced i.p. using large-volume dialysis via a Tenckhoff dialysis catheter. The relative cytotoxicity of Adriamycin to its two major metabolites, adriamycinol and adriamycin aglycone, was also determined in the clonogenic assay. Both derivatives produced suppression of ovarian cancer colony formation; however, Adriamycin was more cytotoxic that was either metabolite. PMID- 7471055 TI - Metabolism in the F344 rat of 4-(N-methyl-N-nitrosamino)-1-(3-pyridyl)-1 butanone, a tobacco-specific carcinogen. AB - The metabolism of the tobacco-specific carcinogen, 4-(N-methyl-N-nitrosamino)-1 (3-pyridyl)-1-butanone (NNK), was studied in the F344 rat, in which it induces tumors of the nasal cavity, liver, and lung. When NNK was incubated with rat liver microsomes and a reduced nicotinamide adenine dinucleotide phosphate generating system, metabolites resulting from alpha-hydroxylation, carbonyl reduction, and N-oxidation were isolated. alpha-Hydroxylation at the methylene carbon gave 4-oxo-4-(3-pyridyl)butanal, whereas alpha-hydroxylation at the methyl carbon gave myosmine and 4-hydroxyl-1-(3-pyridyl)butan-1-one. The formation of these products involved the intermediacy of electrophilic diazohydroxides or carbonium ions which may be proximate or ultimate carcinogens of NNK. Carbonyl reduction gave 4-(N-methyl-N-nitrosamino)-1-(3-pyridyl)butan-1-ol and N-oxidation yielded 4-(N-methyl-N-nitrosamino)-1-(3-pyridyl-N-oxide)-1-butanone. When rats were gavaged with NNK, the microsomal products of alpha-hydroxylation were not detected in the 48-hr urine. Compounds which presumably resulted from further oxidation or reduction of these products, 4-oxo-4-(3-pyridyl)butyric acid, 4 hydroxy-4-(3-pyridyl)butyric acid, and 4-hydroxy-1-(3-pyridyl)butan-1-ol, were isolated. 4-(N-Methyl-N-nitrosamino)-1-(3-pyridyl)butan-1-ol and 4-(N-methyl-N nitrosamino)-1-(3-pyridyl-N-oxide)-1 butanone were also urinary metabolites. PMID- 7471056 TI - Growth of cell colonies in soft agar from biopsies of different human solid tumors. PMID- 7471057 TI - Comparative effects of different strains of Corynebacterium parvum on natural cell-mediated cytotoxicity. AB - Because there are conflicting reports regarding the effects of Corynebacterium parvum (CP) on natural killer (NK) cell activity, several different strains of CP were compared. In replicate experiments, age- and sex-matched mice received 0.25 mg i.p. injections of one of four types of CP; formalin-killed strain 6134; heat killed strain 6134; formalin-killed strain 5888 (actually Corynebacterium granulosum); or formalin-killed CP from the Pasteur Institute. At various days thereafter, two to three mice from each group were sacrificed to determine spleen weight, cellularity, and NK cell activity versus YAC-1 lymphoma cells. The CP from the Pasteur Institute augmented NK cell activity 3 days following injections; however, the activity returned to normal by Day 7 and remained at that level. On the other hand, strain 5888 did not cause as great an increase in lytic activity as did the Pasteur Institute CP at Day 3; but by Day 10 after injection, NK cell levels were above control, and they remained elevated through Day 21. Both the heat-killed and formalin-killed preparations of strain 6134 stimulated NK cell activity initially but resulted in a loss of activity at the later times tested. Experiments done with different doses and routes of injection yielded similar results. Thus, we were able to demonstrate that different types of CP have different effects on NK cell activity in mice and that the general kinetics of these effects were independent of dose or route of administration. PMID- 7471058 TI - Enhancement of antitumor activity of alkylating agents by the radiation sensitizer misonidazole. AB - The influence of the radiosensitizer misonidazole on the effectiveness of several alkylating agents and cis-platinum against advanced solid murine tumors was investigated. Tumor regrowth delay, frequency of tumor regressions, and animal life span were used to evaluate misonidazole in combination with cyclophosphamide, L-phenylalanine mustard, 1-(2-chloroethyl)-3-trans-4 methylcyclohexyl)-1-nitrosourea, aziridinyl-benzoquinone, and cis-platinum. In the advanced M5076 ovarian carcinoma, misonidazole enhanced the activity of cyclophosphamide, L-phenylalanine mustard, 1-(2-chloroethyl)-3-trans-4 methylcyclohexyl)-1-nitrosourea, and aziridinyl benzoquinone, but not cis platinum. In early B16 melanoma, misonidazole plus cyclophosphamide was no more effective than cyclophosphamide alone. In advanced Lewis lung carcinoma, misonidazole enhanced the antitumor activity of cyclophosphamide but not 1-(2 chloroethyl)-3-trans-4-methylcyclohexyl)-1-nitrosourea. Misonidazole, at 1000 mg/kg, increased the antitumor effectiveness of L-phenylalanine mustard and cyclophosphamide in M5076 tumors by factors of 2.2 and 1.8, but caused only a 1.2 and 1.3-fold increase in the myelotoxicity of these agents as determined by spleen colony assay of normal bone marrow. Misonidazole also increased the toxicity of cyclophosphamide and L-phenylalanine mustard in non-tumor-bearing mice but to a lesser degree than it enhanced antitumor activity. These results indicate that misonidazole is capable of enhancing the effects not only of ionizing radiation but of alkylating agents as well. PMID- 7471059 TI - Alkylation of guanosine and deoxyguanosine by phosphoramide mustard. AB - Phosphoramide mustard, an active metabolite of cyclophosphamide, has been reacted separately with guanosine and deoxyguanosine in aqueous solution at pH 7.4. The major adduct which was formed in each case has been isolated by reverse-phase high-pressure liquid chromatography. The structure of the major adduct, as determined by a combination of ultraviolet and field desorption mass spectrometry, is that of phosphoramide mustard, one arm of which has reacted with guanosine or deoxyguanosine in position 7. These adducts are much less stable than was 7-methylguanosine, and they decompose with a half-life of 1.3 hr at 37 degrees and pH 7.4. This instability may contribute to the action of phosphoramide mustard at a molecular level. PMID- 7471060 TI - Adjuvant-antigen requirements for active specific immunotherapy of microscopic metastases remaining after surgery. AB - We studied the conditions required for eradication by immunization of occult lymph node metastases which remained after surgical removal of an intradermally transplanted cavian hepatoma. Guinea pigs that received no postsurgical treatment all died with progressively growing lymph node metastases. The growth of these metastases could be prevented in a significant proportion of the animals by postsurgical treatment with vaccines containing oil-in-water emulsions of Mycobacterium bovis strain Bacillus Calmette-Guerin (BCG) cell walls admixed with live or irradiated tumor cells. Vaccines containing living tumor cells cured most of the guinea pigs but produced tumors at the vaccine sites in a few animals. Irradiated tumor cell vaccines were not tumorigenic but required more tumor cells for successful therapy. Therapy was dependent both on the dose of tumor cells and on that of BCG cell walls. Microgram doses of BCG cell walls were required for a therapeutic effect; milligram doses of BCG cell walls inhibited the therapeutic response. Animals rendered tumor free by postsurgical vaccine therapy rejected an intradermal challenge with living tumor cells. PMID- 7471061 TI - Immunogenic properties of soluble cytosol fractions on Meth A sarcoma cells. AB - Tumor-associated transplantation antigen (TATA) was found to be present in fractions derived from the cytosol of the Meth A cell. Meth A ascites cells were disrupted nuclei and membranes were removed by low- and high-speed centrifugation, and the soluble protein was fractionated by ammonium sulfate precipitation and gel filtration chromatography. The TATA of the soluble cytosol fractions appears to be identical with the TATA solubilized from plasma membranes. The TATA of the cytosol fractions was found to be associated with proteins of an approximate apparent molecular weight of 60,000, specific for the Meth A tumor, and as immunogenic as the membrane-derived TATA. In addition, the most enriched TATA cytosol fraction shows inhibition of an antiserum capable of detecting a tumor-specific surface antigen of Meth A. These results suggest that Meth A TATA is not an integral membrane protein and may be related to the tumor specific surface antigen detected serologically. PMID- 7471062 TI - Cytotoxicity and DNA cross-linking activity of 4-sulfidocyclophosphamides in mouse leukemia cells in vitro. AB - Two sulfido derivatives of cyclophosphamide (CP), 4-S-(hexane-6-ol) sulfidocyclophosphamide and 4-S-(propionic acid)-sulfidocyclophosphamide, were studied for their in vitro cytotoxicity against L1210 cells and for their DNA damaging effects in these cells. These derivatives spontaneously hydrolyze under physiological conditions to form 4-hydroxycyclophosphamide, the active metabolite of cyclophosphamide. The two derivatives were compared with phosphoramide mustard, the presumed alkylating species generated by 4-hydroxycyclophosphamide decomposition, for cytotoxic and DNA cross-linking actions. The three compounds yielded colony survival curves that were similar in shape, but the sulfido derivatives were 4 or 5 times as potent as was phosphoramide mustard. All three compounds produced DNA-protein cross-links as well as interstrand cross-links as measured by alkaline elution. The time course of cross-link formation and removal for the three compounds was similar. The sulfido compounds, however, were 4 to 5 times as potent as was the phosphoramide mustard in the formation of interstrand cross-links, in agreement with the cytotoxicity findings. The higher potency of the sulfido compounds was not attributable to the generation of acrolein. These findings indicate that sulfido derivatives of CP can act directly (without metabolic activation) on cells, probably through spontaneous stepwise conversion to phosphoramide mustard, the presumed proximal alkylating agent. The cell killing effect may be mediated by phosphoramide mustard-induced DNA interstrand cross-linking. Sulfidocyclophosphamide CP derivatives appear to be suitable for in vitro studies of the mechanism of action of CP. Sulfidocyclophosphamide CP derivatives may also have therapeutic potential as CP-like drugs that do not require metabolic activation. PMID- 7471063 TI - Sedimentation of DNA from L1210 cells after treatment with 4' demethylepipodophyllotoxin-9-(4,6-O-2-thenylidene-beta-D-glucopyranoside) or 1 beta-D-arabinofuranosylcytosine or both drugs. PMID- 7471064 TI - Biochemical, pharmacological, and phase I clinical evaluation of pseudoisocytidine. AB - Pseudoisocytidine (psi ICyd) is a C-nucleoside with enhanced stability and resistance to enzymatic deamination when compared to 5-azacytidine and 1-beta-D arabinofuranosylcytosine. Elimination kinetics in plasma using [14C]psi ICyd showed a beta-phase for t1/2 for 14C of 2 hr and a beta-phase t1/2 of unchanged psi ICyd of 1.5 hr. Net recovery of radioactivity in urine over 24 hr varied between 40 and 80% of the administered dose; 50 to 90% was unchanged drug and the rest was pseudouridine. Human leukemic cells in vitro deaminated psi ICyd very slowly, formed appreciable quantities of pseudoisocytidine triphosphate, and incorporated small amounts into RNA and DNA. Clinical trials were done using a daily i.v. injection for 5 consecutive days. Hematological or intestine toxicities were not seen, nor was depression of white blood cell count observed in leukemic patients. Hepatic toxicity proved to be dose limiting; this was characterized by an early phase with elevation of prothrombin time and aspartate aminotransferase. A later phase with cirrhosis was observed in two patients. Autopsy showed massive hepatic necrosis in patients dying of acute toxicity and micronodular cirrhosis in one patient dying with the chronic form. PMID- 7471065 TI - Progressive loss of DNA and lowering of the chromosomal mode in chemically transformed C3H/10T1/2 cells during development of their oncogenic potential. AB - When mouse embryo C3H/10T1/2 fibroblasts are transformed by 7,12 dimethylbenz(a)anthracene, the development of oncogenic potential is associated with characteristic morphological changes, which can be classified into three types and which are also dependent upon the number of cell passages. This paper reports changes in DNA content studied by flow cytometry of ethidium bromide stained nuclei and changes in chromosome number observed by counting 50 randomly chosen Colcemid-arrested metaphases of each cell culture population. The nontransformed cells maintained a stable hypotetraploid DNA content. The morphologically transformed cell showed no changes during their first passages in culture, except for the appearance of small polyploid subpopulations. The occurrence and increase of the oncogenic potential in the higher passages in the transformed cells were associated with decrease in nuclear DNA content as well as reduction in chromosome number. For the most oncogenic passages, the reduction of chromosome numbers was considerable, and there was no overlapping between the chromosome numbers of the original and the most oncogenic cells. This finding supports the view that the transformation process is a result of progression (chromosomal changes and subsequent selection) and not only of the selection of cells that were present at the start of the culture. The genetic material obviously lost in the most oncogenic cells may be different from the DNA material lost by the cells that remained nononcogenic. PMID- 7471066 TI - Erythrocyte production and survival in Rauscher murine leukemia virus-infected BABL/c mice. AB - Red blood cell production in normal and Rauscher murine leukemia virus-infected mice was investigated using 55Fe as a marker. Using autoradiographic techniques, increases in the percentage of labeling of red blood cells were found in blood smears taken at different time intervals after pulse labeling of the erythroid precursor cells. The total reticulocyte production per unit time is more than 2.8 fold in Rauscher murine leukemia virus-infected mice as compared with that in uninfected mice. The life span of the newly formed cells was measured after [51Cr]chromate labeling of transfused erythrocytes in infected and in control mice. The life span was indicated by time that one-half of the labeled erythrocytes disappeared (t1/2) was reduced to one-quarter of that of erythrocytes of uninfected mice. The functioning of the newly formed cells was analyzed by measuring the glucose utilization versus lactate production and by measuring the activities of a number of enzymes involved in glucose metabolism. Comparison of glucose metabolism in normal and leukemic mice and in mice recovering from artificially induced anemia revealed that the metabolic activity of erythrocytes from leukemic mice corresponds to the activity of young erythrocyte populations. The increased reticulocyte production is, apparently, a result of the degree of anemia in infected animals. This anemia is not compensated for, however, since the loss of erythrocytes surpasses the flux of new red blood cells from the hematopoietic organs into the peripheral blood. PMID- 7471067 TI - Clinicopathological study of gastric cancers and precancerous states detected by fetal sulfoglycoprotein antigen screening. AB - A clinicopathological analysis is presented of gastric cancer cases detected in a mass screening trial in Finland, using the oncofetal antigen, fetal sulfoglycoprotein antigen, as a marker. The survey covered a population of 53,020 between the ages of 40 and 70, the percentage of participation being 74.8%. Of these participants, 3,508 subjects (8.8%) were found to be fetal sulfoglycoprotein antigen secretors, and among them 36 gastric cancers, one gastric carcinoid, and 10 tubular adenomas were detected. Both main histological types of gastric cancer, intestinal and diffuse, were represented. There were 15 early cancers. In addition, there were three widely spread superficial cancers. Because of early diagnosis, the prognostic view for these cases is clearly better than that found in the clinic by conventional means, curative resection being carried out in 28 cases (78%). PMID- 7471068 TI - Excision of N-acetoxy-2-acetylaminofluorene-induced DNA adducts from chromatin fractions of human fibroblasts. AB - The excision and persistence of covalent DNA adducts formed by N-acetoxy-2 acetylaminofluorene (AAAF) were studied in human skin fibroblasts. The changes in adduct concentration as a function of posttreatment incubation were measured in purified nucleosomal core DNA and in total nuclear DNA, and from these data the changes in adduct concentration in nucleosomal linker DNA were calculated. Immediately after N-acetoxy-2-acetylaminofluorene treatment which introduced 20 to 38 mumol adducts per mol DNA-P, the adduct concentration was 4 to 5 times higher in linker DNA than in core DNA. Adduct removal was rapid during the first 8 hr of incubation and occurred 3.5 to 4 times more efficiently from linker DNA than from core DNA. After 24 hr incubation, adduct removal continued only at a very low rate, leaving a substantial fraction of adducts unexcised. This fraction of persistent adducts had a value of 0.5 and was independent of the initial adduct concentration in the range of 12 to 115 mumol adducts per mol DNA-P. Approximately 65% of the persisting adducts were located in nucleosomal cores. The initial differences in adduct concentration between linker DNA and core DNA diminished during posttreatment incubation. This was entirely due to preferential early adduct excision from linker DNA. No evidence was obtained for repair induced long-range nucleosomal movement in normal fibroblasts or constitutive movement in the absence of excision repair in xeroderma pigmentosum fibroblasts. Nucleosomal movement would tend to diminish the concentration differences between linker DNA and core DNA. PMID- 7471069 TI - Decreased norepinephrine concentration in normal tissue neighboring a malignant tumor. AB - We observed in five adenocarcinomas that norepinephrine as well as nerve fibers were absent from tumor tissue. The number of nerve fibers in normal tissue neighboring the tumor was normal. In contrast, norepinephrine concentration was decreased in normal tissue immediately surrounding tumor and increased in a stepwise fashion with the distance from tumor. Since there is evidence that some catecholamines possess antitumor activity, we suggest that depletion of catecholamines and perhaps other neurotransmitters in normal tissue neighboring a tumor may be of importance for invasion and accelerated malignant growth. PMID- 7471070 TI - Effect of commercial saccharin preparations on urethan-induced lung tumorigenesis in strain A mice. AB - The effect of commercial saccharin preparations on urethan-induced mouse lung tumorigenesis was assessed by gavaging groups of male strain A mice with 1-g/kg doses of each saccharin preparation on a daily basis 5 days/week. Gavage was initiated 1 week before i.p. injection of either a low (0.1 mg/g) or a high (1 mg/g) dose of urethan and continued until the mice were sacrificed 16 weeks after urethan administration. The average number of surface lung tumors per mouse for each group of mice was determined and was compared statistically with the appropriate control group. The commercial saccharin preparations did not produce an elevated lung tumor response when administered alone. One of the four saccharin preparations enhanced the lung tumor response to urethan when given in conjunction with the low dose of urethan, but this enhancement was not statistically significant. At the high urethan dose, all saccharin preparations produced a statistically significant enhancement of the lung tumor response to urethan. PMID- 7471071 TI - Radioimmunodetection of primary and metastatic ovarian cancer using radiolabeled antibodies to carcinoembryonic antigen. AB - 131I-labeled goat immunoglobulin G (IgG) prepared against carcinoembryonic antigen (CEA) was administered at an average dose of 1.0 mCi (180 to 250 microgram IgG protein) to patients with ovarian tumors in order to evaluate this method of tumor detection and localization, termed the radioimmunodetection of cancer. All primary cancers in 13 patients could be localized, whereas the metastases in six of nine cases could be imaged by external scintigraphy. However, only two of these cases showed metastatic spread by more conventional diagnostic techniques, including computer-assisted tomography, ultrasonography, and angiography. Successful tumor radiolocalization appeared to depend on tumor size, with lesions smaller than 2 cm in diameter not being detected. Tumors containing a CEA concentration above 115 ng/g, including a benign neoplasm, could be localized with radioactive anti-CEA antibodies. Administration of radioiodinated normal goat IgG to four patients with malignant or benign ovarian tumors failed to show tumor radioimmunodetection. One of these cases subsequently demonstrated a 4- x 4-cm tumor after receiving specific radiolabeled anti-CEA IgG. This study shows that ovarian neoplasms containing CEA can be detected and localized by external photoscanning after the application of radioiodinated antibodies to CEA and that, in this small series of patients, primary and secondary tumors could be detected in 100 and 67% of the cases, respectively. PMID- 7471072 TI - Explantation and subculture of epithelial cells from human uterine ectocervix. AB - Human ectocervical explant cultures were grown in medium with D-valine substituted for L-valine. Pure epithelial cell monolayers were obtained with both dialyzed and undialyzed fetal calf serum. Epithelial cell explant colonies grown in D-valine medium supplemented with undialyzed serum could routinely be subcultured if plated at a density of 1.5 x 10(4) cells/sq cm or higher. Such cultures could be passaged at least three times and could yield up to 21 population doublings per culture lifetime. Contaminating fibroblastic colonies were never detected in these cultures, which were free of surface-associated fibronectin as revealed by immunofluorescent tests. Both primary and passaged epithelial colonies retained many characteristic morphological features of ectocervical epithelium when examined by light and electron microscopy. Such cultures may be of use in investigating the action of viral and chemical carcinogenic agents upon epithelial cells in vitro. PMID- 7471073 TI - Chromosomal analysis of human prostatic adenocarcinoma cell lines. AB - Although detailed cytogenetic analysis has been carried out in many types of cancer, there is little information on the chromosomal makeup of prostatic cancer cells. Karyological analyses of cell lines derived from both metastatic and primary prostatic carcinoma have been carried out by Q-, C-, and sequential banding techniques. The metastatic line, PC-3, isolated from a bone marrow specimen, is an established epithelial line which is tumorigenic in nude, athymic mice and forms colonies in semisolid agar suspension. A subline, PC-3/M, was isolated from a PC-3-induced mouse tumor. Karyotypic analysis of PC-3 by Q- and C banding showed the cells to be aneuploid at all culture passage levels. The modal chromosome number shifted from 62 to 55 between the 5th and 50th passages. PC-3 has a unique karyotype. Chromosomes 2, 3, 5, 15, and Y were always absent. At least 11 different marker chromosomes were observed. The subline, PC-3/M, had a similar karyotype and retained the parental PC-3 markers. PC-3/M had a more restricted chromosomal frequency distribution range. Nearly 73% of the PC-3/M cells examined had 60 or 61 chromosomes in contrast to the wide distribution seen in PC-3. Silver staining for nucleolus organizer regions indicated that the number of functional nucleolus organizer regions in PC-3 was proportional to the number of acrocentric chromosomes. Banding analysis of PC-5-PI isolated from primary prostatic adenocarcinoma indicated that this line also had a characteristic karyotype with 28% pseudodiploid and 72% pseudotetraploid components. All metaphases examined were partially trisomic in chromosome 9 and lacked a demonstrable Y chromosome. PMID- 7471074 TI - Antitumor activity of L-canavanine against L1210 murine leukemia. AB - We have made a preliminary assessment of the antitumor activity of the arginine analog, L-canavanine, in leukemic mice. This analog is known to substitute for arginine in protein biosynthesis in many prokaryotic and eukaryotic systems. Previous studies with cells grown in vitro indicated that canavanine caused a marked inhibition of DNA synthesis and viability. The system used in the present study was C57BL/6 x DBA/2 F mice bearing L1210 leukemic cells. Following an i.v. injection of 10 mg canavanine, the t1/2 beta of canavanine in the serum was estimated at 16 min. This finding suggested that frequent injections of high doses of canavanine would be required for an effect on tumor cell proliferation. DNA synthesis by the L1210 cells, assayed by [3H]thymidine incorporation, fell to 9% of the control value after 12 hourly i.p. injections of canavanine (20 mg each). A constant s.c. infusion of 20 mg/hr for 24 hr caused an 86% inhibition of DNA synthesis. The antitumor activity of canavanine was tested against L1210, using a 24-hr infusion schedule with treatment starting 24 hr after i.p. inoculation of 10(5) cells. An optimal dose of 18 g/kg body weight produced a median increased lifespan of 44% (p less than 0.005). These results suggest that L-canavanine may be useful as an antitumor agent. PMID- 7471075 TI - Contrasting effects of subtotal enteric bypass, enterectomy, and colectomy on azoxymethane-induced intestinal carcinogenesis. AB - Compensatory hyperplasia after extensive loss of functioning small or large intestine might predispose to the development of neoplasia in the residual adapted bowel. To test this hypothesis, male Fischer rats were randomized to receive 85 to 90% jejunoileal resection or bypass, subtotal colectomy, or no operation (controls). One week later, the first of six weekly s.c. injections of azoxymethane (15 mg/kg/week) was given. At the 36th week postoperatively, mean body weight after enterectomy or colectomy it was 78 to 79% of control. Adaptation after all three operations was characterized by 22 to 84% increments in villous height and crypt depth in the residual functioning ileum (p = 0.05 to 0.001); the depth of colonic crypts was unchanged. Fewer rats developed intestinal tumors after enteric bypass (36%) than after any of the other treatments (80 to 91%) (p = 0.01 to 0.001); the depth of colonic crypts was unchanged. Fewer rats developed intestinal tumors after enteric bypass (36%) than after any of the other treatments (80 to 91%) (p = 0.01 to 0.001). Compared with controls, bypass reduced the number of colonic tumors by 77% (p less than 0.001). Although resection did not affect colonic tumor yield, it tripled the incidence of tumors in the duodenum and jejunum (p = 0.025). Colectomy promoted rectal carcinogenesis (p less than 0.05). Anastomotic tumors were commoner after intestinal resection. the lower frequency of tumors after jejunoileal bypass contrasts with enhanced carcinogenesis after enterectomy or colectomy. Profound reduction in body weight may prevent the promotional effect of adaptive hyperplasia. PMID- 7471076 TI - Phase I and pharmacological studies of 5-fluorouracil administered intraperitoneally. AB - A Phase I study was conducted of 5-fluorouracil administered i.p. in a 2-liter volume of 1.5% Inpersol. The drug was administered via Tenckhoff peritoneal dialysis catheters to ten patients with tumors confined to the i.p. space. Dialysis concentrations ranged from 5 micro M to mM. Complications of the dialysis procedure alone included mild abdominal discomfort and 2 cases of gram negative bacterial peritonitis, both easily controlled with antibiotics. 5 Fluorouracil caused the same pattern of toxicity as when administered by other routes. There was no local or central nervous system toxicity. Dose-limiting toxicity included pancytopenia and mucositis at a dialysis concentration of 4.5 to 5 mM administered for eight consecutive 4-hr exchanges. There were two documented responses in eight evaluable patients. 5-Fluorouracil concentrations were measured by high-pressure liquid chromatography. Peritoneal fluid concentrations decline in a first-order fashion with a half-life of 1.6 hr. The mean permeability area product was 14 ml/min. A mean of 82% of drug was absorbed in 4 hr. Plasma levels rise over the first 30 to 45 min and decline in a nonlinear fashion. Plasma levels are substantially lower than are peritoneal fluid levels. Mean 4-hr peritoneal fluid concentration was 298 times the simultaneously measured plasma levels. Total body clearance ranged from 0.9 to 15 liters/min and declined with increasing dialysate concentration. We conclude the i.p. route is a relatively safe way to deliver high concentrations and large amounts of drug to the i.p. cavity with a significant pharmacological advantage over conventional routes of administration. PMID- 7471077 TI - Transplantation of primary plasma cell tumor without 2,6,10,14- tetramethylpentadecane (pristane) treatment of the hosts. AB - Small inocula of primary plasma tumor cells which do not transplant i.p. unless recipients are conditioned with pristane do so readily when recipients are given i.p. injections of a peritoneal exudate induced by pristane inoculating, but free of pristane. PMID- 7471078 TI - Excision and postreplication DNA repair capacities, enhanced transformation, and survival of Syrian hamster embryo cells irradiated by ultraviolet light. AB - The frequency of ultraviolet light (UV)-induced neoplastic transformation of Syrian hamster embryo cells (HEC) is enhanced 3- to 10-fold when the cells are first treated with either X-irradiation or with methyl methanesulfonate. Maximum enhancement occurs when the interval between the two treatments is 48 hr. The relevance of UV-induced transformation to neoplasia is confirmed because the transformation to neoplasia is confirmed because the transformants produce tumors when injected into nude mice. Excision and postreplication DNA repair were studied to determine whether the enhanced transformations were associated with either of these repair mechanisms. Independent of X-ray or of methyl methanesulfonate pretreatment, approximately 25% of the pyrimidine dimers are excised within 24 hr in cells irradiated with UV with 3 J/sq m. During this period, more than 70% of the genome of cells irradiated with UV has been replicated. Postreplication repair is measured by the time required to chase pulse-labeled nascent DNA strands to parental-sided DNA. Regardless of pretreatment, 1 and 3 hr are required for pluse-labeled DNA in control and irradiated (10 J/sq m) cells, respectively, to reach parental size. Therefore, no correlation is found between a change in the rate of excision or postreplication repair and enhancement of transformation. Relative to control cloning efficiency, the survival of HEC contain more than 10(5) pyrimidine dimers/genome. The level of survival is similar to the survival of human skin fibroblasts which excise pyrimidine dimers four to five times as efficiently. Moreover, postreplication repair cannot account for the ability of these cells to survive because it is three times slower than in human fibroblasts. Therefore, other repair mechanisms must be responsible for HEC survival and transformation. PMID- 7471079 TI - Acinar cell carcinoma of the rat pancreas grown in cell culture and in nude mice. AB - An acinar cell carcinoma of the pancreas, which was induced in a male Wistar rat by repeated injections of azaserine, was propagated in cell culture. Sheets of epithelial-like cells grew within 2 weeks and were subcultured serially. Initially acinar cell carcinoma in the culture medium produced a high level of amylase, but the secretion ceased rapidly as cells began to proliferate. Only negligible amounts of trypsinogen and chymotrypsinogen were detected in cell homogenates at passages 7 and 9. The chromosome distribution ranged from hypodiploid to hypertetraploid. When cultured cells were transplanted s.c. into nude mice, palpable tumors appeared within 4 weeks and could be transplanted serially. Histological examination of the tumor showed poorly differentiated carcinoma without acinar structures. Tumor homogenate contained amylase, trypsinogen, and chymotrypsinogen, and the electron microscopic examination revealed that many tumor cells contained zymogen-like granules. These results indicate that pancreatic acinar cell carcinoma in cell cultures, in which there was no differentiated function, can be activated to synthesize tissue-specific enzymes when transplanted into nude mice by yet undefined factors present in the host animals. The cell line and transplantable tumors of pancreatic acinar cell carcinoma may be useful in the analysis of the biological behavior of this type of tumor and in the study of the control mechanisms of the synthesis of tissue specific products in cells. PMID- 7471081 TI - Morphogenesis of neoplasms induced in the hamster trachea with N-methyl-N nitrosourea. AB - The tracheas of 95 male Syrian golden hamsters were locally exposed to a flowing stream of a 1% solution N-methyl-N-nitrosourea using a specially designed cannula at weekly intervals for 15 weeks. Animals were killed at specified times from 9 through 27 weeks following the first dose. Epithelial flattening was observed in over 90% of the tracheas throughout the study, but the proportion of the epithelium involved decreased from 100% at 9 and 16 weeks to 25 to 50% at 27 weeks. Hyperplastic foci of squamous metaplasia were found in 5% of the tracheas at 9 weeks, 90% of tracheas at 16 weeks, 83% of tracheas from 18 to 26 weeks, and 80% of tracheas at 27 weeks. Dysplastic foci of hyperplasia were observed in 5% of the tracheas at 9 weeks, 90% of the tracheas at 16 weeks, 17% of tracheas from 18 to 26 weeks, and 1% of tracheas at 27 weeks. Neoplasms were found in low incidences (2%) at 16 weeks. Papillomas and carcinomas were present in 13 and 42% of the tracheas from 18 to 26 weeks and 30 and 75% at 27 weeks, respectively. Dysplastic foci, as well as small focal carcinomas which had invaded the tracheal cartilages, were frequently found in close association with invaginations of the epithelium into the submucosa. PMID- 7471080 TI - Antitumor activity of cytidine dialdehyde and its effects on nucleotide and nucleic acid synthesis. AB - Cytidine dialdehyde inhibited the growth of leukemia L1210 cells in culture at a 50% inhibitory concentration of 3.5 X 10(-5) M and, when administered i.p. at 200 mg/kg daily for 5 days, increased the mean survival of L1210 tumor-bearing mice by up to 171%. Given by the s.c. or i.v. routes, the compound was ineffective. The ethanol adduct of cytidine dialdehyde, although inactive in cell culture, increased the mean survival of L1210 tumor-bearing mice by up to 225% when administered i.p. but was inactive upon s.c. administration. Exposure of L1210 cells in culture for 25 hr to cytidine dialdehyde at the 50% inhibitory concentration increased the ribonucleoside di- and triphosphate pools, slightly increased deoxyadenosine triphosphate, deoxythymidine triphosphate, and deoxyguanosine triphosphate pools, and caused a pronounced increase in the deoxycytidine triphosphate pool. As determined by the rate of pyrimidine precursor incorporation into nucleic acids, this concentration of drug showed no effect on RNA synthesis but caused a reduction in DNA synthesis to 53% of control. Exposure of L1210 cells for 3 to 6 hr to 10(-4) M cytidine dialdehyde, a concentration which inhibits growth completely, effected an increase in the ribonucleoside di- and triphosphate pools and a rapid decrease of the deoxythymidine triphosphate pool. The deoxycytidine triphosphate and deoxyguanosine triphosphate pools decreased more slowly, and the deoxyadenosine triphosphate pool remained slightly elevated. Analysis of the rate of substrate incorporation into nucleic acids showed that this concentration of drug produced an 80% decrease in RNA synthesis and a 75% decrease in DNA synthesis 3 hr after drug exposure. These results suggest that the mechanism of action of cytidine dialdehyde may be due to its initial interference with DNA synthesis followed by a generalized inhibition of DNA, RNA, and protein synthesis at cytotoxic concentrations. PMID- 7471082 TI - Comparisons of liver transfer RNA methyltransferase and adenosylmethionine decarboxylase activities of male and female rats. AB - The activities of liver transfer RNA (tRNA) methyltransferases from control or ovariectomized female rats were found to be higher than those of control or castrated males. Administration of testosterone to ovariectomized females caused activity to decrease to the level of the males. Conversely, administration of estrogen to castrated males resulted in liver enzyme levels similar to those of the females. When the substrates for in vitro methylation were either mixed heterologous tRNA's from Escherichia coli or mixed homologous methyl-deficient tRNA from livers of ethionine-treated rats, the difference in activity between males and females was about 35%. When amino acid-specific tRNA's from E. coli were used as substrates, the ratios of activity of enzymes from females to that of males were: tRNANfMet 1.5; tRNAMetMet 1.1; tRNASer3 1.85; tRNAPhe 1.1; and tRNATyr 1.25, indicating that there are qualitative as well as quantitative differences in the liver tRNA methyltransferases of the two sexes. The adenosylmethionine decarboxylase activity of female rat liver preparations was approximately double that found for males. Testosterone, given to ovariectomized females, lowered the activity of this enzyme to about the same level as that of males. It is not clear whether the observed sex-related differences in activity of several adenosylmethionine-utilizing liver enzymes represent isolated phenomena or are indicative of a sex-related difference in the rate of liver adenosylmethionine turnover. PMID- 7471083 TI - Effect of cholesterol content on antitumor activity and toxicity of liposome encapsulated 1-beta-D-arabinofuranosylcytosine in vivo. AB - 1-beta-D-Arabinofuranosylcytosine (ara-C) was encapsulated in anionic multilamellar liposomes prepared with different lecithin: cholesterol (L:C) ratios. The chemotherapeutic activity of encapsulated ara-C was compared with comparable doses of ara-C in 0.85% saline solution (single- and multiple-dose schedules) in mice bearing L1210 (i.p.) leukemia. Maximum survival was obtained in animals given injections of ara-C (40 mg/kg) encapsulated in liposomes with a L:C ratio of 1:1. The effect of L:C ratio on survival was not pronounced in multiple-dose schedules. Multiple doses (every 4.5 hr for 3 separate injections) of 40 mg/kg with L:C ratios of 1:1 and 1:0.5 were toxic, resulting in 83 and 50% mortality, respectively, of mice by Day 7. This study shows that drug efflux and in vivo antitumor activity and toxicity of encapsulated ara-C is influenced by the cholesterol content of the liposomal lipid bilayer. PMID- 7471084 TI - Mutagenicity and tumor-initiating activity of cyclopenta(c,d)pyrene and structurally related compounds. AB - The biological activities of benzo(a)pyrene, cyclopenta(c,d)pyrene, and 12 other structurally related compounds were assessed by mutagenicity studies with bacterial and mammalian cells and/or skin tumorigenicity studies with mice. The ability of the parent hydrocarbons to be metabolically activated to mutagenic products was examined in strains TA98 and TA100 of Salmonella typhimurium, using 3 experimental protocols. In each case, cyclopenta(c,d)pyrene was metabolically activated to products mutagenic to the bacteria to a greater extent than was benzo(a)pyrene. However, 7,8-dihydrobenzo(a)pyrene and 0,10-dihydrobenzo(e)pyrene were the best substrates for metabolic activation to bacterial mutagens. Highly purified epoxide hydrase added to a purified and reconstituted monooxygenase system readily abolished the mutagenic activity observed in strain TA100 of S. typhimurium when cyclopenta(c,d)pyrene was the substrate, but not when benzo(a)pyrene was the substrate. Inherent mutagenicity of several epoxides of the hydrocarbons generally paralleled the ability of their potential metabolic precursors to be activated to mutagens. 1-Pyrenyloxirane and 10,11 dihydrocycloheptapyrene 8,9-oxide were highly mutagenic in strains TA98 and TA100 of S. typhimurium, and in the former strain these activities were comparable to that observed with 9,10-epoxy-7,8,9,10-tetrahydrobenzo(a)pyrene, 4-Pyrenyloxirane was significantly less mutagenic than was 1-pyrenyloxirane in both strains of bacteria and in mammalian cells. Benzo(a)pyrene was over 20 times more tumorigenic than was cyclopenta-(c,d)pyrene, and it was the most potent of the 11 compounds tested for tumor-initiating activity in 2-stage initiation-promotion experiments on the skin of mice. Cyclopenta(c,d)pyrene had tumor-initiating activity comparable to that of benzo-(a)anthracene, but it was significantly less active than chrysene. Thus, contrary to inferences made from its high mutagenic activity, cyclopenta(c,d)pyrene is a weak tumor initiator on mouse skin. PMID- 7471085 TI - Shortened platelet survival as a cause of thrombocytopenia in mice with L1210 leukemia. AB - Thrombocytopenia is a frequent complication of acute leukemias of humans and animals. To define the possible causes of this decrease in platelets, we have studied platelet kinetics in mice after transplantation of 10(6) ascites cells from mice bearing L1210 leukemia. The circulating half-time of 51Cr-labeled platelets was reduced to approximately one-half that of controls when studied 1 or 3 days posttransplantation. Recovery of transfused 51Cr-labeled platelets was reduced to approximately one-half that in controls when studied 3 days after introduction of L1210 cells. Megakaryocyte concentration showed no change during the 5-day survival after i.v. infusion of leukemic cells but was increased on Day 5 and i.p. inoculation with an average host survival of 7 days. Megakaryocyte diameter distributions were significantly shifted toward larger sizes beginning on Day 2 after i.v. inoculation and on Day 3 after i.p. inoculation. Twenty-four hr [3H]thymidine labeling indices of megakaryocytes were significantly increased beginning on Day 3 after i.v. inoculation but were significantly decreased on Days 5 and 6 after i.p. introduction of L1210 cells. We conclude that the decrease in platelets in mice transplanted with L1210 leukemia results primarily from shortened platelet survival and organ pooling. Megakaryocytes remain normal in concentration but increase in size, a usual response to decreases in platelet count. PMID- 7471086 TI - DNA synthesis, mutagenesis, DNA damage, and cytotoxicity in cultured mammalian cells treated with alkylating agents. AB - The effects of N-methyl- and N-ethyl-N'-nitro-N-nitrosoguanidines and methyl and ethyl methanesulfonates on DNA synthesis in C3H/10T 1/2 mouse embryo fibroblasts and Chinese hamster V79 cells have been analyzed by measuring the incorporation of tritiated thymidine into the trichloroacetic acid-insoluble fraction and the purified DNA of the cells, and by measuring the fraction of parental DNA replicated by equilibrium density sedimentation in neutral cesium chloride gradients. The inhibition of DNA synthesis produced by the alkylating agents at less than or equal to 90% lethal dose was uniformly associated with the cytotoxicity of all four of the agents in both lines of cells. The molar concentrations of the compounds required to inhibit DNA synthesis or produce cytotoxicity also correlated well in a log-log plot, with their potencies in producing DNA lesions that are associated with mutagenesis. However, equitoxic doses of the alkylating agents that inhibited DNA synthesis to equal extents produced frequencies of mutations and of alkali-labile DNA lesions that differed by about an order of magnitude, which did not produce major departures from the linear correlation apparent in the above log-log plot. Therefore, the kinds of DNA lesions that inhibit DNA synthesis and are associated with cytotoxicity in cells treated with the above alkylating agents are not uniformly associated with alkali-labile DNA lesions and gene mutations. Thus, evaluations of mutagenic potency, by assays of DNA synthesis and cytotoxicity, will be falsely high for compounds that predominantly produce DNA lesions that inhibit DNA synthesis and are associated with cytotoxicity. Such evaluations will be falsely low for compounds that predominantly produce DNA lesions that are mutagenic. PMID- 7471087 TI - Photochemically induced binding of aflatoxins to DNA and its effects on template activity. AB - The covalent binding of photoactivated aflatoxin B1 ((AFB1) to DNA and its effect on template activity have been investigated. AFB1 in its ground state complexes more preferentially with denatured DNA than with native DNA. The covalent linkage between 3H-AFB1 and DNA under near-ultraviolet light irradiation shows 1 AFB1 per 529 Pi of calf thymus DNA. The photoaddition also induces a conformational change of the DNA, as detected by circular dichroism. Because the photobinding of AFB2 to DNA is negligible (1 AFB2 per 5485 Pi), we suggest that the 8,9-C=C bond is the major binding site of AFB1. The AFB1-DNA adduct shows a substantial inhibition of its template activity for DNA synthesis (by 52%) and RNA transcription (by 74%) in vitro. Since both AFB1 and AFB2 themselves had little inhibitory effect on the activities of DNA and RNA polymerases, the alteration of DNA by photoactivated AFB1 and AFB2 is responsible for the dramatic reduction of template activity in DNA and RNA synthesis. Short-chain polynucleotides retained by type VS membrane filter (0.025-micrometers pore size) indicate that premature chain termination occurred in transcribing the AFB1-DNA and AFB2-DNA adducts as the result of disrupted movement of the enzymes along the DNA chain. PMID- 7471088 TI - DNA repair in human cells containing photoadducts of 8-methoxypsoralen or angelicin. AB - Photoactivated 8-methoxypsoralen (8-MOP) has been proven to be clinically effective for a number of dermatological conditions including lichen planus, mycosis fungoides, and psoriasis. 8-MOP forms two types of covalent photoproducts with DNA, monoadducts, and bifunctional adducts which cross-link the two DNA strands. Angelicin is a congener of 8-MOP which forms only monoadducts. We have used the combined density and isotopic labeling technique to study repair replication in cultured human fibroblasts treated with either of these compounds and exposed to near-ultraviolet light. In human diploid fibroblasts (WI-38), the time course of repair replication for both compounds is similar. Drug concentration and ultraviolet dose responses are also similar for 8-MOP and angelicin. No repair replication was stimulated by either compound in xeroderma pigmentosum cells from Complementation Group A (XP12BE). These results suggest that repair replication in response to 8-MOP is primarily a response to monoadducts and that the enzymatic pathway for this repair synthesis shares at least one step with the pathway for repair of pyrimidine dimers. Cross-link persistence in treated cells was assayed by use of the single-strand-specific S1 nuclease to digest DNA that did not renature readily following heat denaturation. Partial removal of cross-links was observed in normal, xeroderma pigmentosum variant, and Fanconi's anemia fibroblasts, but not in xeroderma pigmentosum Group A cells. PMID- 7471089 TI - Flow analysis of DNA content and cell size in non-Hodgkin's lymphoma. AB - Cellular DNA content, Coulter volume, and light scatter were measured in cell suspensions from 30 non-Hodgkin's lymphomas in order to assess flow analysis as a quantitative and reproducible means of evaluating these diseases. Nonneoplastic control populations included 31 samples obtained from lymph nodes, spleens, tonsils, and peripheral blood. Cellular DNA and light scatter were measured on ethanol-fixed cells by flow microfluorometry using nuclei isolated from chicken erythrocytes as an internal standard. For DNA analysis, the cells were stained with propidium iodide following RNase treatment. The cellular DNA content of the human populations was expressed as a ratio between the DNA content of the human G0-G1 cells and that of the chicken erythrocyte nuclei. The mean DNA ratio for the 31 nonneoplastic samples was 2.83 +/- 0.08 (S.D.) In these samples, the coefficient of variation of the human G0-G1 peak ranged from 2.27 to 3.63% (mean 3.09 +/- 0.32%). Fifteen of 30 non-Hodgkin's lymphomas, including 7 of 15 low grade lymphomas and 8 of 15 high-grade lymphomas, had abnormal DNA content, the majority containing hyperdiploid G0-G1 populations. In six malignant lymphomas with normal DNA content, the coefficient of variation of the human G0-G1 peak, corrected for differences in instrument setting was greater than that seen in the nonneoplastic populations. A good correlation between the percentage of cells calculated to be in the S phase of the cell cycle and the expected clinical behavior of the tumors was observed. In those lymphomas in which the S-phase percentages could be calculated, 11 of 13 low-grade lymphomas had less than 5% of the cells in S phase, and 7 of 10 high-grade lymphomas had greater than 5% of the cells in S phase. Thirteen of 21 neoplastic cases in which Coulter volume determinations were performed could be distinguished from the nonneoplastic controls on the basis of their modal volume. Although some correlation was observed between light scatter of ethanol-fixed cells and Coulter volume measurements on unfixed cells, light scatter was found to be less discriminatory. Altogether, by all three flow parameters studied, 26 of 30 (87%) of the neoplastic cases could be distinguished from nonneoplastic controls. PMID- 7471091 TI - Pharmacokinetics of vincristine in the cerebrospinal fluid of subhuman primates. AB - Adult rhesus monkeys were given 2-mg/sq m i.v. bolus injections of radiochemically pure tritiated vincristine (VCR). Simultaneous specimens of blood and cerebrospinal fluid (CSF) were sampled from 5 min to 72 hr following injection. CSF was obtained from Ommaya reservoirs which had been implanted s.c. in each monkey. VCR and its metabolic and/or decomposition products rapidly entered the CSF producing low concentrations in this fluid; 5 min following injection, the concentration of VCR was 2.3 nM, whereas approximately a 100-fold greater concentration (203.8 nM) was attained in the plasma. Throughout a 3-day period of observation, constant, low levels of VCR and its metabolic and/or decomposition products (2.3 to 5.7 nM) were maintained in the CSF. The principal form of VCR present in the CSF 1 hr or more after i.v. injection was a water soluble metabolite and/or decomposition product(s). Although the levels achieved in the CSF are unlikely to be lethal to tumor cells, the finding of persistent low concentrations of VCR and its metabolic and/or decomposition products in the CSF indicates prolonged exposure of neural tissue which might result in neurotoxicity, particularly cumulative neurotoxicity following multiple doses of VCR. PMID- 7471090 TI - Inhibition of growth of colorectal carcinoma in nude mice by monoclonal antibody. AB - Hybridoma-derived monoclonal anti-colorectal carcinoma antibodies suppressed the growth of colorectal carcinoma in nude mice as evidenced by a lower incidence of tumors, a longer latency period, and a smaller volume of tumors in antibody treated than in control animals. The growth-inhibiting properties of monoclonal anti-colorectal antibodies seem to be specific for colorectal carcinoma cells. This is indicated by the lack of effect of the antibodies on the growth of melanomas or bronchogenic carcinomas and by the binding of the antibodies in vivo to colorectal carcinoma cells but not to lung or kidney cells from tumor-bearing animals or to other tumor cells implanted in other animals. Inhibition of tumor growth was most probably mediated by antibody-dependent cell-mediated cytotoxicity. The results of these studies could provide an approach to the study of immunotherapeutic possibilities for anti-colorectal carcinoma antibodies in humans. PMID- 7471092 TI - Saccharin-induced hyperplasia of the rat urinary bladder. AB - Dietary administration of sodium saccharin to male Fischer rats increased the rate of DNA synthesis of the urinary bladder epithelium as measured by tritiated thymidine uptake. A multifocal mild epithelial hyperplasia was induced, which was most readily detected by scanning electron microscopy, with pleomorphic microvilli present on many of the hyperplastic cells. PMID- 7471093 TI - Cell kinetic-directed sequential chemotherapy with cyclophosphamide and adriamycin in T1699 mammary tumors. AB - The present studies were initiated to investigate the changes [3H]deoxythymidine labeling index, primer-dependent DNA polymerase labeling index, and S-G2 transition after treatment of T1699 transplantable mouse mammary tumors with Adriamycin (5 mg/kg) and cyclophosphamide (100 mg/kg). Treatment with these agents resulted in intervals of subnormal tumor cell proliferation as indicated by decreased [3H]deoxythymidine labeling index, primer-dependent DNA polymerase labeling index, and S-G2 transition. Recovery, as indicated by increases in [3H]deoxythymidine labeling index, primer-dependent DNA polymerase labeling index, and S-G2 transition, was observed three days after Adriamycin treatment and six to seven days after cyclophosphamide treatment. To evaluate the predictive nature of the kinetic changes for effective time sequencing, sequential combination chemotherapy protocols were designed and tested in T1699 tumor-bearing mice. The results from these studies showed that the most effective chemotherapy schedules were those in which the drugs were sequenced to coincide with the cell kinetic recovery from the single agents alone. These effective sequencing intervals were also found to be effective when used in multifraction sequential combination chemotherapy protocols. The results suggest that changes in cell kinetic parameters following drug perturbation can provide indications as to potentially efficacious as well as nonefficacious sequencing intervals. PMID- 7471094 TI - Relative activities of thymidylate synthetase and thymidine kinase in rat tissues. AB - The activities of thymidylate synthetase and thymidine kinase were compared in tissues of normal (adult and developing), cortisol-injected, and tumor-bearing rats. The purpose of the study was to determine whether the activities of these two enzymes, which catalyze reactions leading to the same metabolic intermediate, changed proportionately, reciprocally, or independently under different physiological conditions. Both enzymes had high activities in fetal tissues. Thymidine kinase concentrations decreased shortly before or immediately after birth; in several tissues, transient postnatal peaks in thymidine kinase activities appeared within the first 3 weeks after birth. Thymidylate synthetase activities declined gradually after parturition and showed no significant postnatal rises. In sucklings given injections of cortisol, thymidine kinase activities were reduced substantially in eight tissues while thymidlyate synthetase decreased only in lung and thymus of 11-day-old rats. In tumor-bearing rats, thymidine kinase activity increased dramatically in spleen, whereas thymidylate synthetase activities only doubled. In host liver, rises in thymidine kinase activities were not always matched by increases in thymidlyate synthetase. In the tumors, both activities were higher than in most normal adult tissues. Despite the differential sensitivities of the two enzymes to cortisol and tumor bearing, thymidylate synthetase and thymidine kinase were closely correlated in tissues of untreated animals. The Spearman rank correlation coefficient for 112 tissues was 0.895, while the correlation coefficient between the standard scores of the activities was 0.839. The activities of the two enzymes did not appear to be reciprocal or compensatory during normal differentiation or during dedifferentiation associated with tumor bearing, but their potentials for activity were independent of each other. PMID- 7471095 TI - Radiation synergism by repair-inhibiting nitrosoureas in L1210 cells. PMID- 7471096 TI - Relationships between carcinogenesis in vivo and alkylation and solvolysis in vitro. AB - Carcinogenic bromomethyl- and chloromethylanthracenes and benz(a)anthracenes were found to react rapidly in vitro with DNA under physiological conditions and to varying degrees with nucleotides and nucleosides. The compounds were assayed for production of lung adenomas in strain A mice and relative carcinogenic potencies were compared with in vitro alkylation and with in vitro solvolysis rates. Success at quantitative correlation with carcinogenic potency was not obtained with either in vitro parameter. However, comparable noncarcinogenic compounds lacked their marked in vitro reactivity with DNA. Some potential polynuclear electrophiles, including chloromethyl derivatives of acridine and benz(c)acridine, a hydroxymethyl and an acetoxymethyl derivative of benz(a)anthracene, an acetoxymethylanthracene, and an acridinylglycine ester, were found not to be carcinogenic in the lung adenoma test. PMID- 7471097 TI - Nonrandom abnormalities in chromosome 1 in human testicular cancers. AB - Trypsin G banding was performed on metaphase chromosomes from 14 cell lines derived from primary tumors or metastases of 11 patients with testicular cancer. Most of the cell lines, 11 of 14, have a modal number between 51 and 61. All lines have numerical and structural changes involving chromosome 1 with trisomy of the q arm being the common aberration. Break points in chromosome 1 were nonrandom, being concentrated in the regions of p12, q12, p36, and p22, which resulted in morphologically identical marker chromosomes in different cases. These changes probably are not artifacts of cell culture. In one instance, three lines derived from the same patient, one from tissue removed at operation, and two from separate metastases removed at autopsy nearly 3 years later after unsuccessful radiotherapy and chemotherapy had identical chromosome compositions. In another case, lines derived from a primary tumor and a metastasis from the same patient also had identical marker chromosomes. The consistent involvement of chromosome 1 in aberrations may be associated with the highly malignant nature of testicular cancers. PMID- 7471098 TI - Clinical phase I-II and pharmacokinetic study of high-dose thymidine given by continuous intravenous infusion. AB - High-dose thymidine (dThd) was given to 12 patients with advanced hematological and solid tumors. The dose schedule used was 75 g/sq m/day, given i.v. continuously for 5 days or more. Myelosuppression, especially leukopenia, was the dose-limiting toxicity. Nonhematological toxicities affected the gastrointestinal tract (nausea, vomiting, anorexia, diarrhea, and indigestion) and the central nervous system (somnolence, headache, visual illusions, and memory impairment). Patients who had received cumulative doses of dThd developed alopecia. Thymine crystals were noted in the urine after refrigeration. Tumor regression (less than partial remission) occurred in one patient with melanoma. Three of four patients with acute leukemia had a fall in peripheral white blood cell counts and blasts but no marrow improvement. Four patients with adenocarcinoma (three colon, one unknown primary) had stable disease. Pharmacokinetic studies revealed that, at a dThd dose of 75 g/sq m/day, millimolar concentrations of dThd and thymine can be achieved in the plasma. The half-life of dThd was approximately 100 min. One third of the plasma concentrations was measurable in the cerebrospinal fluid. dThd was mainly excreted by the kidneys. PMID- 7471099 TI - Resistance patterns of Walker carcinosarcoma 256 and other rodent tumors to cyclophosphamide and L-phenylalanine mustard. AB - Comparison is made of the development of resistance to cyclophosphamide (CPA) and L-phenylalanine mustard (L-PAM), of cross-resistance, and chromosome counts, in Walker 256 (W256), rat sarcoma R3 (R3), leukemia L1210, and Ridgway osteogenic sarcoma. For development of resistance the single maximum tolerated doses of CPA or L-PAM were used, each for two sublines in the four tumors. In W256 after only one to five treatment generations, all sublines were resistant, whereas only by generation 10 had R3/CPA, R3/L-PAM, and L1210/CPA reached marked resistance, and L1210/L-PAM reached moderate resistance. All four Ridgway osteogenic sarcoma sublines were essentially still as sensitive as the parent tumor. Long established resistant sublines from previous studies (greater than 20 treatment generations) were used for cross-resistance, chromosome, and stability studies. All W256-resistant sublines were cross-resistant to CPA, L-PAM, and thiotepa; but the sublines of the other tumors, although showing marked, or in the case of L1210/CPA, complete resistance to their respective inducing agents, retained moderate-to-full sensitivity to the other alkylators. W256/CPA and W256/L-PAM were mainly polyploid (greater than 80% of cells), whereas the other tumors were mainly diploid or near diploid. During 10 to 20 untreated generations the degree of drug resistance remained unchanged in W256 and L1210 lines, but was reduced in R3 and Ridgway osteogenic sarcoma lines. The resistance pattern of W256 appears to be compatible with a simple selection mechanism, whereas those of the three other tumors suggest involvement of multiple determinants. This study suggests that some, but not all, tumors have universal cross-resistance between different types of alkylating agents. PMID- 7471100 TI - Acyltransferase-mediated binding of N-hydroxyarylamides to nucleic acids. AB - N-Hydroxyarylamides are metabolically activated to nucleic acid-binding species by the action of N,O-acyltransferase (AT). The substrate specificity of these enzymes in rat, guinea pig, monkey, baboon, pig, and human liver has been examined by measuring the AT-mediated nucleic acid binding of the N-formyl, N acetyl, and N-propionyl derivatives of N-hydroxy-2-aminofluorene. Human and pig enzymes catalyzed binding in the order formyl greater than acetyl greater than propionyl, while for the other species the order was acetyl greater than propionyl greater than formyl. Ammonium sulfate fractionation of the cytosols suggested that the baboon and rat have at least two different AT's: one with a higher specificity for the formyl derivative; the other with a marked preference for acetyl and propionyl compounds. Only one form, with a high formyl group specificity, was detected from human liver. The identity of the in vitro AT mediated DNA adducts from rat, baboon, and human liver was established. In each instance, one adduct accounted for greater than 75% of the bound 2-aminofluorene (AF) residues. This product had a high-pressure liquid chromatography retention time and pH-dependent partition characteristics identical to those of an adduct synthesized by an acid-dependent (pH 4.6) reaction of N-hydroxy-2-aminofluorene with calf thymus DNA. This synthetic adduct has been identified as N (deoxyguanosin-8-yl)-2-aminofluorene by nuclear magnetic resonance, mass, and ultraviolet light spectroscopy. Moreover, it was identical to the product obtained from the alkaline (pH 12) hydrolysis of N-(deoxyguanosin-8-yl)-2 acetylaminofluorene. Since an arylaminated (i.e., aminofluorene) residue(s) is the major product found in rat liver DNA following administration of N-hydroxy-N acetyl-2-aminofluorene, these data suggest that AT may play a major role in the formation of this DNA-carcinogen adduct. PMID- 7471101 TI - Mutagenic and carcinogenic metabolites of the carcinogen 15,16-dihydro-11 methylcyclopenta[a]phenanthren-17-one. AB - Microsomal metabolites of the carcinogen 15,16-dihydro-11 methylcyclopenta[a]phenanthren-17-one (Structure I) were separated by high pressure liquid chromatography, and their structures were established on the basis of their ultraviolet and mass spectra, together with considerations of their general chemical properties. This was assisted by comparisons with metabolites formed in the same way from the synthetic 15-hydroxy (Structure III), 16-hydroxy (Structure II), and 11-hydroxymethyl (Structure IV) derivatives, which themselves occur as metabolites of Structural I. Products derived from attack at the two benzo-ring double bonds occurred, but no K-region products were found. Only metabolites having a non-bay region 3,4-dihydrodiol system were mutagenic and bound to DNA after in vitro microsomal activation, and it was concluded that the 3,4-dihydro-3,4-diol (Metabolite e) was the main form and that the 3,4-diols of the monools (Structure II to IV) were minor proximate forms of this carcinogen. In a two-stage experiment, the synthetic 16-ol (Structure II) was shown to be almost as carcinogenic as was Structure I itself in mice; the 15-ol (Structure III) and 11-hydroxymethyl derivative (Structure IV) were much less active. The same order was also observed in the mutagenicity of these compounds in the Ames test. PMID- 7471102 TI - Immunological quantification by high-affinity antibodies of O6 ethyldeoxyguanosine in DNA exposed to N-ethyl-N-nitrosourea. AB - Three immunological methods [radioimmunoassay (RIA), enzyme-linked immunosorbent assay, and radioimmunosorbent technique] were established for quantification of the potentially mutagenic O6-ethyldeoxyguanosine (O6-EtdGuo) in DNA treated with the carcinogen ethylnitrosourea in vivo or in vitro. To obtain high-affinity antibodies for specific detection of low levels of O6-EtdGuo in small amounts of DNA (cells), different schemes were applied for immunization of rabbits with the hapten O6-ethylguanosine coupled to various carrier proteins (rat serum albumin, bovine serum albumin, keyhold limpet hemocyanin). Low-dose immunization with the hapten-keyhold limpet hemocyanin conjugate resulted in antibodies with an affinity constant of 1 to 2 X 10(10) liters/mol and very low levels of cross reactivity with normal as well as other alkylated DNA components. The RIA (the most sensitive of the three assays) detects 0.05 pmol of O6-EtdGuo at 50% inhibition of tracer (O6-ethyl[8,5'-3H]-3'-deoxyguanosine)-antibody binding. This permits quantification by RIA of O6-EtdGuo at an O6-EtdGuo:2'-deoxyguanosine molar ratio of approximately 3 X 10(-7) in a hydrolysate of 100 micrograms of ethylated DNA. By chromatographic separation of O6-EtdGuo prior to the RIA, this value can be lowered to less than 5 X 10(-8). PMID- 7471103 TI - Adaptation of mass spectrometry for the analysis of tumor antigens as applied to blood group glycolipids of a human gastric carcinoma. AB - Total neutral (nonacidic) glycolipid fractions have been isolated from a gastric adenocarcinoma and the surrounding normal gastric tissue removed at laparotomy of a Blood Group B human individual. Each glycolipid mixture was fractionated by silicic acid column chromatography into ten different partly purified glycolipid fractions. These fractions were analyzed by thin-layer chromatography and tested for Blood Group A and B activity. Blood group B activity was found in several fractions from both tumor and normal tissue. Two of the tumor glycolipid fractions reacted with some batches of commercial anti-A antisera, but other antisera tested did not react. No such Blood Group A reactivity was found in the fractions from the normal gastric tissue. The two Blood Group A-active tumor glycolipid fractions were methylated and methylated-reduced, and these two derivatives were analyzed by mass spectrometry. It was shown that these two fractions were mixtures of glycosphingolipids with five to nine sugars. The dominating glycosphingolipids were blood group Leb and B-like hexaglycosylceramides, a B-similar heptaglycosylceramide with an additional fucose, an H-like heptaglycosylceramide, and a Leb-like octaglycosylceramide. Evidence for small amounts of a Blood Group A-similar heptaglycosylceramide with an additional fucose was also found. The finding of a Blood Group A-similar glycolipid in a fraction which reacts with some anti-A antisera is the first chemical evidence for a heterolog blood group antigen in human cancer which has previously been found by histoimmunological techniques. The clinical significance of this finding is discussed in relation to diagnostic procedures and immunotherapy. PMID- 7471104 TI - Comparative study of the phospholipid composition of plasma membranes isolated from rat primary hepatomas induced by 3'-methyl-4-dimethylaminoazobenzene and from normal growing rat livers. AB - Plasma membranes (PM's) were isolated from primary hepatomas induced in Wistar rats by 3'-methyl-4-dimethylaminoazobenzene, from nonhepatomal regions of the same rat livers, and also from various normal rat livers, including the resting and regenerating livers of adult rats and developing livers of postnatal rats. Phospholipid analyses of these PM preparations revealed the following differences in the hepatoma PM in comparison with those of the PM of normal adult resting livers: (a) decrease in the content of total phospholipids; (b) large increase in plasmalogen content; (c) relative increase of sphingomyelin and ethanolamine phospholipids, and the decrease of choline phosphoglycerides, i.e., decrease of the ratios of choline phosphoglycerides to choline phosphosphingoside and of choline-containing phospholipids to ethanolamine-containing phospholipids; (d) decrease in phosphatidylserine and phosphatidylinositol. The phospholipid composition of the PM's from normal growing livers showed definite decreases in phosphatidylserine and sphingomyelin and increases in ethanolamine phospholipids; however, no significant alteration was observed in plasmalogen content in comparison to the PM of normal adult resting livers. The PM from the nonhepatomal regions of the hepatoma-bearing livers did not show those differences observed in hepatoma PM. PMID- 7471105 TI - X-ray sensitivity of fifty-three human diploid fibroblast cell strains from patients with characterized genetic disorders. AB - The in vitro response of 53 human diploid fibroblast strains to X-irradiation was studied using a clonogenic survival assay. The strains, derived from patients with a variety of characterized clinical conditions, most with a genetic component, ranged in Do (a measure of the slope of the survival curve) from 43 to 168 rads. The mean Do's of six strains from normal individuals was 140 to 152 rads, with an overall range, based on the extremes of their standard errors, of 128 to 164 rads. Three-quarters of the strains studied fell within this range. Strains identified as sensitive came from patients with ataxia telangiectasia, progeria, the two genetic forms of retinoblastoma, and partial trisomy of chromosome 13. No marked radiosensitivity was found among strains derived from patients with a number of other conditions associated with a predisposition to malignancy. PMID- 7471106 TI - Survey of radiosensitivity in a variety of human cell strains. AB - gamma-Ray sensitivity for cell killing was assayed in 54 human cell strains, including some derived from individuals suffering from certain heritable diseases. The overall range of Do values in this study was 38 to 180 rads, indicating a considerable range of variability in humans. The normal sensitivity was described by a range of Do values of 97 to 180 rads. All ten ataxia telangiectasia cell strains tested proved radiosensitive and gave a mean Do value of 57 +/- 15 (S.E.) rads, and these represent the most radiosensitive human skin fibroblasts currently available. Representative cell strains from familial retinoblastoma, Fanconi's anemia, and Hutchinson-Gilford progeria occupied positions of intermediate sensitivity, as did one of two ataxia telangiectasia heterozygotes. Six xeroderma pigmentosum cell strains together with two Cockayne's syndrome cell strains (all known to be sensitive to ultraviolet light) fell into the normal range, indicating an absence of cross-sensitivity between ultraviolet light and gamma-irradiation. PMID- 7471107 TI - Inhibition of RNA synthesis in murine ependymoblastoma by the combination of amphotericin B and 1-(2-chloroethyl)-3-cyclohexyl-1-nitrosourea. AB - The aim of this study was to clarify the mechanism of the potentiation by amphotericin B (AMB) of 1-(2-chloroethyl)-3-cyclohexyl-1-nitrosourea (CCNU) antineoplastic effects on s.c. murine ependymoblastoma. The effect of AMB on tumor cell permeability to CCNU labeled on the cyclohexyl moiety was studied. The radioactivity measured in ependymoblastoma 1, 6, 14, and 25 hr after i.m. injection of 10.4 microCi of 1-(2-chlorethyl)-3-[cyclohexyl-1-14C]cyclohexyl-1 nitrosourea per mouse was significantly higher (p less than 0.001) in the tumors of animals treated with AMB (25 mg/kg 10 hr prior to [14C]CCNU) as compared to controls. The effects of AMB and CCNU given separately or in combination on RNA and protein synthesis were studied by measuring the incorporation of [3H]uridine and [14C]leucine, respectively, into RNA and proteins. The administration of AMB (25 mg/kg) or CCNU (10 mg/kg) did not affect the incorporation of [3H]uridine measured 2 hr after the i.p. injection of 40 microCi of labeled precursor per mouse. On the other hand, the incorporation of [3H]uridine was significantly (p less than 0.001) inhibited in animals treated with AMB (25 mg/kg) followed 10 hr later by CCNU (10 mg/kg), as compared to animals receiving CCNU alone. The inhibition, which reached a maximum of about 35% 24 hr after the administration of CCNU, was not observed when AMB was given after CCNU. The inhibition of RNA synthesis was also observed in mice treated with AMB and cyclohexyl isocyanate (5.4 mg/kg), a degradation product of CCNU. Measurements of [14C]leucine incorporation showed that AMB did not increase the inhibition of protein synthesis produced by CCNU. These observations suggest that AMB increases the uptake of a cyclohexyl derivative arising from the degradation of CCNU. The increased uptake of this compound results in inhibition of RNA synthesis. This mechanism could account for the potentiation of the CCNU therapeutic effect produced by AMB, at least in murine ependymoblastoma. PMID- 7471108 TI - Effect of anticancer agents on directional migration of malignant C3H mouse fibroblastic cells in vitro. AB - Invasion of malignant cells was considered as a target for therapy. The effect of various anticancer agents, which were known to permit or to prevent invasion in vitro, on the growth and on the directional migration of virally transformed malignant C3H mouse fibroblastic cells (MO4) was examined. The increase of the diameter of spheroidal aggregates of MO4 cells in individual shaker culture was used as an index of growth. The mean diameter of the circular area covered by cells migrating from an aggregate explanted on glass, the number of cells in the periphery of this area, the height of the central part of the aggregate, the microcinephotographic aspect of migrating cells, and the immunocytochemistry of the cytoplasmic microtubular complex were considered as indices of directional migration. Ionizing radiation (5,000 and 20,000 R), and 5-fluorouracil (0.1, 0.5, and 1 microgram/ml), known to permit invasion, inhibited growth but allowed directional migration. Nocodazole (0.1 and 1 microgram/ml), known to prevent invasion, interfered with both growth and directional migration. These observations showed that various agents which affected the growth of aggregates of MO4 cells had different effects on the directional migration of these cells; they suggested that proliferation and migration were unrelated cellular activities. The assay for directional migration of cells from a spheroidal aggregate explanted on glass is proposed for the screening of potential antiinvasive agents. PMID- 7471110 TI - Thermal sensitivity of murine tumor cells and hyperthermic tumor eradication. PMID- 7471109 TI - Human embryonal carcinoma grown in athymic mice and in vitro. AB - Tissue from 19 human testis tumors was transplanted into athymic mice. One embryonal carcinoma, ECCS, grew rapidly, and this tumor was studied both as a xenograft and an in vitro culture of xenograft-derived tumor cells. Xenografts showed no evidence of differentiation. The embryonal carcinoma cells were heteroploid and showed alkaline phosphatase activity. When tumor cells from the xenografts were grown in vitro, the cells formed aggregates resembling embryoid bodies with epithelium-like cells in the periphery. Regularly, another population of mouse cells which showed several criteria of malignancy overgrew the culture and could be subcultured continuously. These abnormal cells may result from an in vivo or in vitro transformation of mouse stromal cells. PMID- 7471111 TI - Clinical pharmacologic effects of thymidine plus 5-FU. AB - Ten patients with advanced cancer were given 24 courses of thymidine (TdR) plus 5 FU. In the initial part of the study, TdR was given by continuous infusion at a dose of 8 g/m2/24 hrs x 5 days, and 5-FU was given as an iv bolus daily x 5. During the continuous TdR infusion, TdR serum levels ranged from 10(-6) to 10(-5) M, and thymine serum levels ranged from 10(-5) to 10(-4) M. 5-FU serum decay on Day 1 was rapid, and serum decay on Day 5 was slightly prolonged in two of three patients studied. The optimal dose of 5-FU with continuous infusion of TdR seemed to be between 200 and 400 mg/m2 daily x 5 in heavily pretreated patients. In the second part of the study, rapid infusion of TdR at a dose of 8 g/m2/2 1/2 hrs followed by 5-FU resulted in markedly prolonged 5-FU serum decay (initial serum half-life, approximately 240 mins) and definite increase in clinical toxicity. The dose-limiting toxic effect of both schedules was myelosuppression. Administration of TdR in one tenth of the initial infusion time resulted in a nearly 100-fold increase in peak TdR levels. Thymine was the major metabolite of TdR. 5-FU did not influence TdR blood levels. The protracted 5-FU serum decay appears to account at least in part for the increased toxicity of 5-FU when administered following TdR. PMID- 7471112 TI - Toxicity of spirogermanium in mice and dogs after iv or im administration. AB - Toxicity of single-dose spirogermanium was evaluated after iv and im administration to CDF1 mice and beagle dogs. The im LD50 in mice was approximately threefold greater than the iv LD50. The lethal dose in dogs was the same for both routes of administration, but death was delayed after im injection. Convulsive seizures occurred only after the im doses that were lethal, but they were observed after administration of iv doses that were nonlethal. Microscopic evidence of drug toxicity (necrosis and degeneration) was found in mitotically active tissues: intestinal tract, lymphoid tissue, and bone marrow. Necrosis, hemorrhage, edema, and granulation tissue were observed in the muscle injection site. PMID- 7471113 TI - Radionuclide angiography to predict patient response to hepatic artery chemotherapy. PMID- 7471114 TI - Phase I study of 6-diazo-5-oxo-L-norleucine (DON). AB - We conducted a phase I study of 6-diazo-5-oxo-L-norleucine given iv on a twice weekly schedule. Twenty-six evaluable patients received 31 courses of the drug. Doses ranged from 100 to 500 mg/m2. Nausea with vomiting was the dose-limiting toxic effect, transient thrombocytopenia was seen frequently, and mucositis occurred in 39% of the patients. No definite therapeutic responses were observed in 18 patients with measurable lesions. The recommended dose for phase II studies is 200-300 mg/m2 iv twice weekly. PMID- 7471116 TI - Variability in DNA distributions of human neuroblastomas after cyclophosphamide. AB - Major changes in DNA distributions of two human neuroblastoma cell lines growing in vitro and in athymic nude mice occurred after treatment with cyclophosphamide. Pulse treatment of LA-N-1 cells in vitro with liver S-9-activated cyclophosphamide (10 micrograms/ml) caused approximately 50% cytotoxicity; flow microfluorometric analysis of surviving cells demonstrated an increased proportion of G2 + M cells and a decreased proportion of G1 cells, particularly at 48 hrs. Even though LA-N-1 tumors in nude mice did not regress after one dose of cyclophosphamide (250 mg/kg), the percent of G2 + M cells increased and the percent of G1 cells decreased 4-6 days after treatment; the percent of cells in S increased at 2 and again at 8 days. SK-N-MC cells were affected differently by cyclophosphamide. Pulse treatment of these cells in vitro with liver S-9 activated cyclophosphamide caused greater than 85% cytotoxicity and nearly complete elimination of cells in G2 + M at 24 and 48 hrs. Likewise, SK-N-MC tumors in nude mice regressed greater than 50% after cyclophosphamide, and the proportion of G2 + M cells decreased markedly 2-6 days after therapy. We conclude that cyclophosphamide can have different cytotoxic and cytokinetic effects on neuroblastomas. In addition, marked cytokinetic effects may occur even though changes in tumor size are minimal or not detectable. PMID- 7471115 TI - Clinical pharmacology of IMPY by radioimmunoassay. AB - A radioimmunoassay for IMPY has been developed using [3H]acetyl-IMPY and antibody induced by immunizing rabbits with succinyl-IMPY bovine serum albumin conjugates. The method was reproducible and sensitive at a dose of 0.2 microgram/ml. The cross-reactivities of IMPY and IMPY derivatives with the antibody were compared. Using this radioimmunoassay, clinical pharmacology was performed in cancer patients during phase I trials. Following 30-60-minute iv infusions of IMPY (450 1500 mg/m2/day for 5 days), the drug disappeared rapidly from plasma with an initial half-life of less than 10 minutes and second half-life of 4 hours. The half-lives are not affected by minimally abnormal hepatic function and are not dose-related. The apparent volume of distribution was approximately 125 ml/kg. The results obtained on Day 5 were essentially the same as those obtained on Day 1. IMPY was also detected in glioblastoma, brain, and temporalis muscle when the drug (500 mg/m2) was infused iv 4-5 hours before surgery. The tissue drug levels were comparable to or higher than those of plasma. The drug was also measurable in kidney and liver tissues obtained 4 hours after death in a patient who died 21 hours after iv infusion of IMPY at a dose of 1500 mg/m2/day for 3 days. PMID- 7471117 TI - Lactic acidosis associated with metastatic breast carcinoma. AB - Occurrence of lactic acidosis with adequate tissue oxygenation (type B lactic acidosis) has been described in association with leukemia, lymphoma and a single case of Hodgkin's disease. No cases of this type have been reported in association with solid tumors. A case of type B lactic acidosis in a woman with rapidly progressing metastatic breast carcinoma is described. PMID- 7471118 TI - Dianhydrogalactitol and neural tumors: an in vitro, in vivo preclinical evaluation. AB - The effects of dianhydrogalactitol on human neuroblastomas in vitro and in vivo as heterotransplants in nude mice were determined. Four neuroblastoma lines and three primitive neuroectodermal tumor lines were found, in vitro, to have different sensitivities to the drug. The most sensitive in in vitro assays was the neuroblastoma line SK-N-Mc. Tumors from patients resistant to cyclophosphamide were sensitive to dianhydrogalactitol in vitro and in nude mice. The lack of increased cytotoxicity in vitro with concentrations greater than 12 micrograms/ml and the lowered degree of weight loss in mice treated with 6 mg/kg/day x 5 consecutive days compared to 15 mg/kg/day x 2 days (either in sequence or with a 3-day interval) suggest that clinical trials with 5-day courses or constant infusions may be more effective than intermittent pulsed doses. PMID- 7471119 TI - Phase I study of 3-deazauridine in the treatment of adults with solid tumors. AB - A phase I trial of the uridine analog 3-deazauridine was undertaken in 44 adults with solid tumors. The drug was given as a 5-day continuous infusion repeated every 3-4 weeks. The dose-limiting toxic effect was granulocytopenia. Patients with prior nitrosourea therapy or extensive irradiation also had significant thrombocytopenia, and the lowest dose tested, 800 mg/m2/day, was excessive for this group. Mucositis was occasionally severe and was particularly marked in previously irradiated areas. Nausea was mild to moderate. There were isolated episodes of rash, headache, chest pain, and blurred vision. For patients without extensive prior therapy, the recommended dose is 1000 mg/m2/day. No complete or partial remissions were noted. PMID- 7471120 TI - Phase I trial of PALA. AB - PALA was given iv on a weekly schedule to 32 patients with advanced malignant tumors. A course of treatment consisted of three weekly doses. Patients were treated at eight dose levels, ranging from 900 to 6750 mg/m2. Gastrointestinal toxicity (diarrhea) and skin rash were dose-limiting. No consistent myelosuppression occurred, and no renal or hepatic toxicity was observed. Two patients who did not have metastatic intracranial disease experienced episodes of encephalopathy and seizures. No major therapeutic responses were observed during this phase I trial; however minor responses (greater than or equal to 25% tumor decrease) occurred in three patients (one with adenocarcinoma of the lung and two with epidermoid carcinoma of the bladder). Doses of 4500 mg/m2/week for patients with a performance status greater than or equal to 70 and 3750 mg/m2/week for patients with a lower performance status are recommended for phase II studies. PMID- 7471121 TI - Thymidine arrest and synchrony of cellular growth in vivo. AB - Thymidine (TdR) has been used to study the kinetics of in vitro cell proliferation and is currently being used clinically as a single agent at high doses. We have explored the in vivo cytokinetic effects of TdR on rapidly proliferating cell populations by continuous infusions in rats. The nucleoside was lethal at high doses when serum levels approached 10(-2) M.. At levels of 10( 3) M, TdR exposure for greater than 24 hours resulted in bone marrow hypocellularity and peripheral myelosuppression. Pathologic findings were also noted in the intestinal mucosa. Serum TdR levels of 10(-4) M were sufficient to induce arrest of cell growth in S phase by inhibition of DNA synthesis. Subsequent release after TdR exposure produced partial synchronization of the bone marrow and intestinal mucosa cell populations, as shown by microfluorometry and labeling studies to monitor DNA synthesis. A similar arrest of cell cycle traverse has been demonstrated in a tumor cell population by infusing rats bearing transplantable subcutaneous myeloblastomas. The inhibition of DNA synthesis, as determined by labeling studies, was comparable for bone marrow, intestinal mucosa, and myeloblastoma at serum TdR levels of 10(-3) M. This arrest of myeloblast cell growth was dependent on tumor burden and did not effect survival when maintained during 72-hour infusions. The continuous infusion of TdR provides an approach for studying cell kinetics in vivo, and findings similar to those described here have been observed in our clinical studies. PMID- 7471123 TI - Effect of microbial fractions and vehicle on survival of mice bearing Lewis lung carcinoma. AB - Mice bearing pulmonary metastases of the Lewis lung carcinoma were treated iv with a nonviable microbial vaccine following amputation of the primary inoculation site. The vaccine consisted of BCG cell wall skeleton, trehalose, dimycolate, and endotoxin attached to mineral oil microdroplets. Single and repeated doses ranging from 15 to 675 micrograms were tested. Survival was modestly but consistently prolonged by vaccination. A portion of the activity appeared to be due to the Tween saline-oil vehicle. A single low dose (15 micrograms) was as efficacious as higher or repeated doses. PMID- 7471122 TI - Indomethacin treatment of rats with dimethylhydrazine-induced intestinal tumors. AB - Intestinal tumors were induced in male Sprague-Dawley rats following the administration of five weekly doses of 1,2-dimethylhydrazine (DMH) by gavage. At intervals thereafter, groups of rats were given indomethacin (IND) in the drinking water. In three trials, the incidence of rats with tumors was significantly reduced by 40% below the control rats which did not receive IND and there was no significant difference in body weights. Also, in those IND-treated rats which developed tumors, the tumors were generally smaller in numbers and sizes compared to the control rats. A group of DMH-treated rats was treated with crude IND by gavage and the differences between them and untreated control rats were not significant. It is likely that the treatment was directed at the tumors and not at the DMH which induced them. PMID- 7471125 TI - Doxorubicin and ricin, a strongly synergistic combination in mouse leukemia. AB - The effect of combinations of doxorubicin (Adriamycin) and the cancerostatic plant lectin ricin on L1210 leukemic cells in mice was assessed by end-point dilution and spleen-colony assays. The combinations, which were well-tolerated by the mice, acted synergistically on leukemic cells in liver, spleen, and brain. In the bone marrow, a dramatically synergistic effect was obtained with combinations that had only slight effect on the survival of normal stem cells. PMID- 7471124 TI - Phase II study of L-asparaginase in the treatment of pancreatic carcinoma. AB - L-Asparaginase was given to ten patients with advanced nonresectable pancreatic carcinoma because of the demonstration of in vitro sensitivity of the tumor to the drug. No therapeutic value was demonstrated for L-asparaginase. Toxicity was significant, mainly as evidenced by increasing mental confusion and early signs of a coagulopathy. On the basis of this limited study, L-asparaginase seems to have no value in advanced pancreatic carcinoma. The usefulness of L-asparaginase as primary therapy in patients with less advanced disease remains to be determined. PMID- 7471126 TI - Phase II trial of Baker's antifol in metastatic renal cell carcinoma: a Southwest Oncology Group Study. AB - Fifteen patients with metastatic renal cell carcinoma were treated with Baker's antifol at a dose schedule of 250 mg/m2 iv for 3 days every 3 weeks. One objective response was seen, and toxicity was minimal. No usefulness of Baker's antifol in the treatment of metastatic renal cell carcinoma was demonstrated. PMID- 7471127 TI - The antithrombin-binding sequence of heparin studied by n.m.r. spectroscopy. PMID- 7471128 TI - Controlled-release diethylpropion hydrochloride used in a program for weight reduction. AB - In a double-blind, placebo-controlled evaluation in obese adults given comparable dietary and exercise recommendations, controlled-release diethylpropion hydrochloride promoted significantly more weight loss than matching placebo. The mean reduction in 12 weeks was 15.9 lb (average 1.32 lb/week) for 12 patients taking diethylpropion hydrochloride, 10.0 lb (0.84 lb/week) for 13 taking placebo, and 12.2 lb for 13 taking drug for two four-week periods separated by four weeks of placebo (1.38 lb/week on active drug and 0.30 lb/week on placebo). Amphetamine-like side effects were virtually absent. Diethylpropion hydrochloride is an effective adjunct to caloric restriction in therapy of obesity. PMID- 7471129 TI - A comparison of a three-day and seven-day clotrimazole regimen for vulvovaginal candidiasis. AB - Patients with mycologically proven symptomatic fungal infections of the vagina were treated ina double-blind trial with either one 100-mg clotrimazole vaginal tablet daily for seven days (group I) or two tablets daily for three days (group II). Patients were evaluated at one and four weeks after therapy. The investigator's evaluation of treatment efficacy showed 85% (22/26) success in group II compared with 75% (21/28) in group I (P = 0.46). Both groups had improvement itching, discharge, and vaginal and vulval irritation. No significant (P less than 0.10) differences were seen between groups after therapy. Only three side effects were seen. It is concluded that a three-day course of two clotrimazole vaginal tablet daily is as effective and safe as the previously recommended one tablet daily for seven days, and the shorter therapy is likely to improve patient compliance. PMID- 7471130 TI - Safety and efficacy of feprazone for long-term treatment of rheumatoid arthritis and osteoarthritis. AB - Long-term safety and efficacy of feprazone (4-prenyl-1,2-diphenyl-3,5 pyrazolidinedione), an antirheumatic drug that is well tolerated in the gastrointestinal tract, were assessed in a noncontrolled multicenter trial. Administered at a daily dosage of 600 mg for a mean duration of 114.1 days, feprazone was well tolerated by 43 (77%) of 56 treated subjects. Thirteen patients reported side effects, but only five discontinued treatment. The side effects were generally mild and occurred in the first weeks of therapy. Feprazone was found effective after one month on both considered indices of disease activity: Ritchie index (articular index for assessment of joint tenderness) and corticosteroid consumption. PMID- 7471131 TI - Double-blind, placebo-controlled, crossover trial of propranolol as treatment for menopausal vasomotor symptoms. AB - A trial of propranolol for the treatment of vasomotor symptoms was conducted among 37 menopausal women in a double-blind, placebo-controlled, crossover fashion. When compared with placebo, propranolol had a statistically significant effect on the reduction of both the frequency and severity of vasomotor symptoms (P less than 0.01). In selected patients, propranolol may be an alternative to the treatment of vasomotor symptoms with exogenous estrogens. PMID- 7471132 TI - Colchicine in the treatment of Paget disease of bone: a new therapeutic approach. AB - Paget disease of the bone (PDB) has been treated effectively with various agents including calcitonins, diphosphonates, and mithramycin. Each agent has relatively serious toxic side effects or practical inconveniences associated with its use. An effective agent with fewer adverse reactions and a more convenient route of administration would be preferable. The purpose of this study was to evaluate the effect of colchicine for the treatment of PDB in five patients. All patients were symptomatic and had typical changes on roentgenograms and bone scans consistent with the disease. Serum alkaline phosphatase ranged from 408 to 1,311 mU/ml (normal, 30 to 115 mU/ml), and urinary excretion of total hydroxyproline ranged from 68 to 205 mg/24 hr (normal, 30 to 65 mg/24 hr). Colchicine, 0.6 mg, was given orally three times a day to each patient, who had subsequent follow-up with clinical and laboratory determinations evaluated at each visit. The duration of follow-up was eight to 28 weeks, with a mean of 20 weeks. Pain was relieved in all patients, and two became asymptomatic. Serum alkaline phosphatase decreased 18% to 38%, and urinary hydroxyproline decreased 26% to 53% from the pretreatment values. The biochemical values and and clinical symptoms changed markedly in two patients, correlating with withdrawal and reinstitution of colchicine. These results indicate that colchicine may be effective in the treatment of PDB. Although the mode of action and long-term efficacy of colchicine in this disorder remains to be evaluated, the antimitotic effect on osteoprogenitor cells, the adherence of colchicine to the microtubular structures in preexisting osteoblasts, and the nonspecific anti-inflammatory effect of this agent may explain the therapeutic response noted in this study. PMID- 7471133 TI - Relative speed of onset of the antidepressant effect of maprotiline. AB - In a double-blind clinical trial conducted in general practice, maprotiline was compared with amitriptyline. Thirty-three of 40 patients completed the trial, 19 on maprotiline and 14 on amitriptyline. The Hamilton Depression Rating Scale was administered on days, 0, 7, 14, and 21, and a global rating scale of improvement was administered on day 21. Although there were significant reductions in the Hamilton scores by day 21 in both groups, with no significant difference between the compounds, maprotiline produced significantly greater improvement by day 7 than did amitriptyline. A review of published double-blind clinical amitriptyline or imipramine for treatment of depression supports our finding that maprotiline has a faster onset of action than the standard tricyclic antidepressants. PMID- 7471134 TI - First-cousin marriage and sickle-cell anaemia. PMID- 7471135 TI - The significance of tuberculin immunity testing in cancer patients. PMID- 7471136 TI - Chemoarchitectonics of the pons of Taphozous melanopogon (a microchiropdteran bat). PMID- 7471137 TI - Effects of monoamine oxidase inhibitors on release of dopamine and 5 hydroxytryptamine from rat striatum in vitro. PMID- 7471139 TI - Simultaneous detection method of amine and acetylcholinesterase containing nerve fibers in pial vessels--II. Electron microscopic study. PMID- 7471138 TI - Simultaneous detection method of amine and acetylcholinesterase containing nerve fibers in pial vessels--I. Light microscopic study. PMID- 7471140 TI - Electron microscopic visualization of tetramethyl benzidine reaction product on horseradish peroxidase neurohistochemistry. PMID- 7471141 TI - [Investigation of endocrine cells in the duodenum of chicken embryos (author's transl)]. PMID- 7471142 TI - [Alcoholic fermentation: influence of the culture conditions on inhibition by ethanol (author's transl)]. PMID- 7471143 TI - Collagen fibers in enlarged basement membranes in human schistosomal liver and spleen. PMID- 7471144 TI - [Survival of 51Cr homologous platelets and HLA compatibility (author's transl)]. PMID- 7471145 TI - [HLA and oligophrenia (author's transl)]. PMID- 7471146 TI - [The LISS test and its potential uses in routine work (author's transl)]. PMID- 7471147 TI - [Multiple myeloma treatment with adriablastin and BCNU (author's transl)]. PMID- 7471148 TI - [Social significance of alimentary tract diseases in the Czech Socialist Republic - some trends and proposed solutions to the present-day situation (author's transl)]. PMID- 7471149 TI - [Assessing the extent of myocardial infarction in clinical practice (author's transl)]. PMID- 7471150 TI - [Leg blood flow in reactive hyperaemia (author's transl)]. PMID- 7471151 TI - [Levels of circulating immunocomplexes in the sera of patients with diffuse fibrotic interstitial pneumonia (author's transl)]. PMID- 7471152 TI - [Essential choline phospholipids and their effect on HBsAg and selected biochemical tests in cirrhosis of the liver (author's transl)]. PMID- 7471153 TI - Differences and similarities in adjuvant chemotherapy in mouse and man. AB - The effectiveness of adjuvant chemotherapy is easily demonstrated in many mouse models, whereas in man the response is good but far less spectacular. With the aid of a mathematical model the following variables in which differences may exist between mouse and man were tested: 1. Uniformity of tumour load in population 2. Cell kinetic factors in drug sensitivity 3. Variation of drug sensitivity among tumours 4. Drug penetration into tumour 5. Tumour antigenicity 6. Variation in growth rate among tumours It is concluded that the factors 1 and 3 are responsible for the difference. Mouse models are more sensitive by selection of an effective drug and an optimal tumour load. Other factors seem not to affect the results. PMID- 7471154 TI - A method for measuring the generation time and length of DNA synthesizing phase of clonogenic cells in a heterogenous population. AB - Information on the cell cycle of progenitor cells in haemopoietic tissue is useful for understanding population control under physiological and abnormal conditions. Unfortunately, methods that have been developed for measuring cell cycle parameters are applicable only to cells of homogenous populations and not to morphologically non-recognizable progenitor cells such as colony forming units (CFU) that are present at low frequency in a heterogenous population. To circumvent this difficulty, a method was developed to measure CFU cell cycle parameters based on specific killing of cells in S phase by [3H]thymidine ([3H]TdR). This was done by estimating the number of CFU killed following exposure of the cell suspension to [3H]TdR for various time periods. since cycling CFU are continuously entering S phase, a linear curve relating the percentage CFU-kill to the length of exposure of the cells to [3H]TdR in culture can be obtained. The slope of the curve (percentage kill/hr) indicates the rate that CFU enter the S phase and travel through the cell cycle. The inverse of this value will then represent a time period for CFU to move through a complete cell cycle (generation time). The length of S phase can then be obtained by multiplying generation time by the fraction of cells in S phase at time zero. This method has been used to measure generation time and length of S phase of three kinds of haemopoietic progenitor cells: mouse granulocyte-macrophage CFU, human T lymphocyte CFU and CFU from regenerating mouse spleens. This method should be applicable to any normal or neoplastic clonogenic cell populations and the latter could be either of haematological or of solid tumour origin. PMID- 7471155 TI - Differentiation of murine marrow megakaryocyte progenitors (CFUm): humoral control in vitro. AB - Differentiation of mouse marrow megakaryocyte progenitors (CFUm) was studied in vitro by a colony assay using a plasma clot system. Erythropoietin (EPO) from sheep plasma (6 units/mg protein) in doses from 1 to 5 units/ml induced a linear increase in CFUm to a maximum of 20 colonies/10(5) cells plated. Human urinary EPO also induced a dose-responsive increase in CFUm, but the maximum was 9 colonies/10(5) with 2.0 units/ml of EPO and there was a decrease in colonies above that concentration. Thrombocytopoiesis-stimulating factor (TSF) derived from human embryonic kidney culture supernatant fluids induced a dose-responsive increase in CFUm in concentrations from 0.01 to 0.32 mg protein/ml in the absence of added EPO. TSF did not support the growth in vitro of erythroid colonies from mouse marrow (CFUe and BFUe) indicating the absence of EPO activity. In these studies sheep EPO appeared more effective in supporting CFUe growth than human EPO. TSF also had a stimulatory function in megakaryocyte differentiation at a precursor level. Multiple humoral factors play a role in megakaryocytopoiesis in vitro. PMID- 7471156 TI - Rat C-6 glioma cells maintained in an organ culture system: a study of kinetic parameters. AB - Rat C-6 glioma cells were grown on a sponge foam matrix in an organ culture system and the cell cycle parameters, including the growth fraction (GF), were assessed after autoradiography. The zones of growth consisted of a compact upper layer (UL) at the gaseous interface, a central necrotic layer and a deeper lower layer (LL) which invaded the matrix. The fraction of continuously labeled mitoses (FCLM) was similar in both the UL and LL cells. The derivatives of the FCLM curves obtained in three experiments gave an average modal TG2 of 5 hr. A mathematical model relating GF, TG2, TC and labeling index as a function of time, LI(t), was devised for cells in a steady state exposed continuously to tritiated thymidine and was applied to data obtained from UL cells. A mean GF of 9% (range: 8-10%) and a mean cell cycle time (TC') of 27 hr (range: 13-47 hr) were obtained. The mean TS was calculated to be 11 hr (range: 8-16 hr) by the method of grain counts per mitotic figure or grain index (GI). Knowledge of TS permitted alternative calculation of the cell cycle time from the equation Ts/TC = LI(0)/GF:this gave a mean cell cycle time (TC") of 29 hr (range: 20-45hr). Except for the GF, the cell kinetics were comparable to those of the same cell line grown in monolayer culture. The GF in the in vitro system described is in the lower range reported in some human malignant gliomas in vivo. PMID- 7471157 TI - On the derivation of cell kinetics from histomorphology, observed in the rat incisor odontoblast. AB - A method to extract cell kinetic information from histomorphology is presented. Each replicating tissue is essentially an ordered structure with an origin where cells are formed and a periphery toward which they are displaced. The displacement path is called the tissue radius. The tissue variables may be studied in two domains, space and time. The first embraces all the states a cell may assume while the second specifies the cell transition rates. During steady state both domains are related linearly. These ideas are illustrated in the rat incisor odontoblast population whose life expectation is determined by the tooth wall shape. The odontoblast cell population paves the interior of the tooth wall delimiting a cone-shaped pulp. Near the root apex the dentine wall is barely visible. As one proceeds distally, the wall thickens while the pulp narrows. Pulp narrowing is associated with odontoblast cell loss whose magnitude may be deduced from the change of the pulp circumference CI(x) (x is the distance from tooth origin). The odontoblast force of mortality mu(x) may be calculated from the instantaneous perimeter change:mu(x) = - CI'(x)/CI(x); where CI'(x) stands for the derivative of CI(x). This equation serves for the construction of the odontoblast life table which may be studied in space and time. PMID- 7471158 TI - An in vitro model of hematopoietic injury in chronic hypoplastic anemia. AB - Mice treated with high-dose busulfan develop a 'latent' form of bone marrow failure characterized by near-normal peripheral blood counts and marrow cellularity, but marked reductions in marrow pluripotent stem cells (CFUs) and myeloid progenitor cells (CFUc). Spleen cell suspensions from control and 'latent' mice were placed in liquid culture in the presence of colony-stimulating activity. Cells were harvested at intervals up to 14 days and sub-cultured in agar to assay for CFUc. Baseline splenic CFUc did not differ significantly between control and 'latent' mice. Splenic CFUc from control mice increased 50 fold and reached a peak at day 10 in liquid culture. In contrast, splenic CFUc from 'latent' mice increased only 7-fold and reached a peak at day 3. Our results indicate that although splenic CFUc are present in normal numbers in 'latent' mice, their proliferative capacity is markedly reduced, either as the result of defective CFUc self-renewal or defective feed-in from CFUs or both. PMID- 7471159 TI - Detailed kinetic analysis of Shay chloroleukaemia cell population propagated in permanent suspension culture in vitro. AB - Detailed kinetic analysis of a growing cell population is difficult, even when assay conditions are nearly ideal. Therefore, it is usually essential to perform several types of experiments and analyse all the results in terms of a mathematical model, the use of which is not limited a priori by a specified application. In the present study we investigated cell population kinetics using rat chloroleukaemia cells propagated in suspension culture in vitro. The parameters were measured: doubling time of the population, fraction of labelled mitoses, changes in labelling index with time after pulse labelling, continuous labelling and stathmokinetic index. Analysis of the results was based on a computer program CECAM, which is a stochastic model capable of simulating essentially all types of kinetic experiments based on presently known assay techniques. The results showed that precise and reliable information on cell population kinetics could not be obtained from the analysis of any single type of experimental data. In particular, the technique of labelled mitoses underestimated the duration of the G1 phase, owing to subtle label-induced changes in population behaviour. These changes could not have been detected with any certainty without rigorous quantitative comparisons with other types of experimental data. As a whole, however, results obtained by the different techniques were in agreement and the kinetic behaviour of chloroleukaemia cells in vitro could be established in detail. In certain circumstances even minute changes in the kinetic parameters of the cells can modify population behaviour drastically. To study these cases adequately the experiments must be designed with utmost care, preferably with the aid of preceding simulations. This is because demonstration of small primary changes in population kinetics may be beyond the limit of detection of any single assay method. PMID- 7471160 TI - Variation in G1 transit time relative to the cycloheximide and actinomycin D drug restriction points. AB - The transit time distribution at various points in the cell cycle of synchronized Chinese hamster ovary cells was determined from the mitotic index, [3H]thymidine labeling index and increase in cell number monitored at regular intervals after mitotic selection. Variation in G1 transit time compared with that for the total cell cycle indicates that variation in cell cycle transit time occurs mainly during G1 phase. The cycloheximide (5.0 microgram/ml) and actinomycin D (3.0 microgram/ml) restriction points occur 0.2 and 1.7 hr prior to entry into S phase, respectively. The transit time distributions are further characterized by the moments of the distributions. The variance (2nd moment about the mean) of the transit time distribution at the actinomycin D restriction point is similar to the variance of the transit time distribution at the G1/S border, thus variation in cell cycle transit time originates earlier than 1.7 hr prior to entry into S phase (i.e., the first 3/4 of G1). If G1 transit time variability and cell cycle control are related, then the results presented here indicate that the major regulatory events do not occur during late G1 phase. PMID- 7471161 TI - A rapid automated stathmokinetic method for determination of in vitro cell cycle transit times. AB - To provide a rapid method for examining cell cycle dynamics, we utilized continuous exposure of Chinese hamster ovary cells and human colon cancer cells to colcemid to block cycling cells in metaphase, suppressing re-entry into G1. Changes in cell cycle compartment distribution were monitored by DNA flow cytometry. Analysis of the rate of G2 + M compartment accumulation after addition of colcemid permitted calculation of all cycle transit parameters. These compared favorably with data in the same cell lines determined by the fraction of labeled mitoses technique. Serial assessment of DNA flow cytometry after addition of colcemid permits rapid quantitation of cycle traverse rates. PMID- 7471162 TI - Changes in the proliferation characteristics of a solid transplantable tumour of the mouse with time after transplantation. AB - The aim of the present investigation was to study changes in cell proliferation kinetics with time after transplantation in a solid transplantable mammary carcinoma of the C3H mouse. From growth curves and tumour doubling times, three different experimental times were chosen; the exponential phase of growth on the 7th day post-transplantation, during the phase of retardation on the 21st day after transplantation and in the stationary phase of growth on the 30th day after transplantation. It was found that the cell loss factor contributed primarily to the retardation of volume growth. The lowering of the growth fraction played a minor part in the delay of growth while the duration of cell cycle was not prolonged before the plateau phase of growth on the 30th day after transplantation was reached. PMID- 7471163 TI - Genetic control of lipopolysaccharide-induced mobilization of CFUs. Dissociation between early and delayed mobilization of CFUs in complement C5-deficient mice and LPS non-responder mice. AB - Lipopolysaccharide (LPS)-induced mobilization of CFUs from haemopoietic tissues into the circulation has a biphasic pattern. The first rise occurs within 30 min of LPS injection, the second 4-7 days later. This second rise coincides with an increase of the CFUs number in the spleen from about 3000 to about 50,000. We have investigated the relationship between the two peaks by making use of complement C5-deficient mouse strains and the LPS non-responder mouse strains C3H/HeJ and C57BL/10ScCr. These latter two strains lack a serologically identifiable structure ('LPS-receptor') which is present in all LPS-responder strains. After injection of eleven different mouse strains with LPS, the numbers of circulating CFUs increased rapidly in all strains, except in the C5-deficient A/J, AKR/J, DBA/2J and B10.D2/oSn mice. On the other hand, the delayed LPS induced accumulation of CFUs in blood and spleen occurred in all mouse strains tested, including the C5-deficient strains, but not in the LPS non-responder strains C3H/HeJ and C57BL/10ScCr. These results show that (a) early LPS-induced mobilization of CFUs is dependent on the availability of C5, in contrast to the delayed CFUs accumulation in blood and spleen, (b) the presence of the LPS receptor is not required for early CFUs mobilization by LPS and (c) recognition of the mobilizing agent by a specific receptor is required for the delayed accumulation of CFUs in blood and spleen. PMID- 7471164 TI - Regeneration of uterine epithelium after experimental ablation in the rat. AB - Regeneration of the uterine luminal epithelium was studied after its mechanical removal in progesterone-primed rats, leaving one control horn intact. Pulse labelling with [3H]TdR during regeneration, showed a rapid peak of labelling index in remaining glands. A differentiated and highly labelled luminal epithelium reappeared at 34 hr, thereafter showing a rapidly declining LI. After initial depletion, the glandular cell population size was restored within 64 hr, whereas luminal epithelium cell numbers became stabilized at about half normal level. Grain counts after prelabelling showed more rapid dilution in gland cells of stripped uterine horns, indicating accelerated cycling of previously dividing cells. Thymidine labelling indices also showed that, after removal of the epithelium, almost all gland cells became rapidly committed to divide. On average, less than two cell cycles were necessary to restore stable glandular and epithelial population sizes. Numbers of labelled cells were also drastically increased in myometrium and serosa of treated horns. This suggests a non-specific mechanism for stimulation of mitotic activity after ablation of epithelium. PMID- 7471165 TI - Radiotoxicity of incorporated [3H]thymidine as studied by autoradiography and flow cytometry. Consequences for the interpretation of FLM data. AB - The radiotoxic effects of incorporated [3H]thymidine on proliferation kinetics of flash-labelled (30 min, 0.3 microCi/ml, 40 Ci/mM) L-929 cells in vitro were studied by means of autoradiography and flow cytometry. The flow cytometric results obtained by applying the BUdR-33258 Hoechst technique, using new evaluation procedures, showed that the labelled cells are delayed in their progression through the S and G2 + M phases, leading to mitotic delay. From autoradiographs, the fraction of labelled mitoses was determined and, in addition, the ratio of labelled and of unlabelled mitotic cells to all cells. The radiotoxic effects are not evident from the FLM curve, even if the ratio of labelled mitotic cells to all cells shows a highly distorted shape. A mathematical model has been developed that describes the perturbed cell kinetics due to radiotoxic effects of the incorporated [3H]thymidine. These findings have considerable consequences for the interpretation of autoradiographic data, especially of labelled mitoses curves. PMID- 7471166 TI - Haematopoiesis in mice heterozygous for the W trait: defective formation of transient endogenous spleen colonies. AB - It has been determined that W/+ and Wv/+ heterozygous mice, as compared with normal +/+ homozygous littermates, form significantly lower numbers of transient 5-day endogenous spleen colonies in response to X-irradiation. This defect was evident for doses of irradiation between 2-6 Gy (200-600 rad) and was associated with a slightly increased radiosensitivity of the assayed precursor cells (TE CFU) in W heterozygotic mice. Moreover, the defect was transplantable, i.e., intrinsic to the marrow cells and not to the microenvironment, and was not associated with a similar decrease in cells which form erythropoietic bursts in vitro (BFUe). This study provides a cellular basis for increased radiosensitivity of W/+ and Wv/+ mice and suggests that the 'W' mutation is semi-dominant, both with respect to the white spotting and TE-CFU formation. PMID- 7471167 TI - A critique of the practice of comparing control data obtained at a single time point to experimental data obtained at multiple time points. AB - The normal circadian rhythm in DNA synthetic activity (DNA-SA) in the tip of the mouse tongue is presented. When this rhythm, obtained from mice which were not treated (NT) or handled, was compared to the rhythms obtained from mice treated with saline (SAL) or 25 mg/kg isoproterenol (IPR), no alteration in the rhythm was observed after either treatment. The conclusion from this chronobiological, experimental design was that IPR had no effect on DNA-SA in the tip of the tongue. However, when three single time points (08.00, 11.00 or 14.00) are selected from the SAL-treated, control rhythm and compared to the multiple time point data from the IPR-treated mice, three very different, statistically supported conclusions were reached. The common practice of obtaining data at only one time point in control animals and comparing these data to data obtained from drug-treated animals at multiple time points is an example of poor experimental design which results in erroneous conclusions and unnecessary confusion in the literature on in vivo research. PMID- 7471168 TI - Inhibition of glycoconjugate secretion by colchicine and cytochalasin B. An in vitro study of human airway. AB - The effects have been analyzed of cytochalasin B and colchicine on the secretion of glycoconjugates by human bronchial explants labeled in vitro with radioactive glucosamine. Both cytochalasin B and colchicine had no effect on baseline 14C labeled glycoconjugate release but caused a dose-dependent (10(-7)-10(-4) M) inhibition of 14C-glycoconjugate release and discharge of labeled macromolecules from mucous and serous cells induced by 5 X 10(-5) M methacholine. Quantitative autoradiographic analyses showed that neither cytochalasin B nor colchicine inhibited 3H-threonine or 3H-glucosamine incorporation into mucous and serous cells of the submucosal glands or goblet cells of the airway epithelium. Colchicine (10(-5) M) but not cytochalasin B significantly reduced the rate at which labeled macromolecules were transported through mucous, serous and goblet cells but this effect was not observed until 4h after the addition of colchicine. Neither cytochalasin B nor colchicine affected the basal rate of labeled macromolecule discharge from mucous, serous or goblet cells. At a concentration of 10(-5) M, both agents completely inhibited the increase in labeled macromolecule discharge induced in mucous and serous cells by methacholine. Our results suggest that in the submucosal gland of human airways microtubules and microfilaments may be important in secretagogue-induced but not in baseline cellular glycoconjugate discharge, implying that the mechanisms of the two processes differ significantly. Furthermore, a role for microtubules is suggested in the transport of secretory granules through mucous, serous and goblet cells. PMID- 7471170 TI - Similarities and differences in the structure of segmentally homologous neurons that control the hearts of the leech, Hirudo medicinalis. AB - The neural circuit that controls the hearts in the leech comprises an ensemble of synaptically interconnected cardiac motor neurons (HE cells) and cardiac interneurons (HN cells). Both the HE cells and the HN cells constitute segmentally homologous sets. We have investigated the structure of these neurons by iontophoretic injection of Lucifer Yellow dye. Bilateral pairs of HE cells have been identified in segmental ganglia 3-19 of the nerve cord. Their structure was found to be nearly identical from ganglion to ganglion and from animal to animal. Bilateral pairs of HN cells have been identified in segmental ganglia 1-7 of the nerve cord. Their dendritic structure was found to vary from ganglion to ganglion. These segmental differences among HN cells were observed consistently from animal to animal. Some of the segmental differences in HN cell structure correlate with previously described physiological differences. PMID- 7471169 TI - High-voltage electron microscopy of normal and irreversibly sickled red blood cells. AB - A high-voltage electron microscopic study of normal red cells and irreversibly sickled red cells (ISCs) was conducted. Comparison with intact, critical-point dried red cells revealed that the ISC fraction could always be identified because of the presence of numerous echinocytes. Examination of the unsealed ghosts after incubation in 3,3'-diaminobenzidene (DAB) to detect hemoglobin (Hb) bound to the plasma membrane revealed that Hb adhered to the cytoplasmic surface of the ISC membrane. The Hb was concentrated in the surface projections of the echinocytes and also was seen as granules associated with the filamentous substructure of the plasma membrane. The role of this adherent Hb in exerting a transmembrane effect to alter the surface properties of the cell is discussed. PMID- 7471171 TI - Ultrastructural studies on the nucleoli of the follicle cells in the lizards Acanthodactylus scutellatus hardyi and Eremias brevirostris. AB - The ultrastructure of the nuclei in the ovarian follicular cells has been examined in the lizards Acanthodactylus scutellatus hardyi, and Eremias brevirostris. During certain growth stages, the nuclei of the intermediate and the pyriform cells become large and conspicuous, and usually contain numerous morphologically variable nucleoli. The latter may be nucleolonemal, compact, ring shaped, "filamentous", "fine-filamentous", "amorphous" or "course-granular" in type, and the number and type in each nucleus seems to be related to the developmental stage. The type of nucleolus may indicate different phases of activity in the nucleus and may also suggest an association with the production of different types of RNA. PMID- 7471173 TI - Multiple effects of 5 mM sodium butyrate on Physarum polycephalum macroplasmodia. AB - In a Physarum polycephalum macroplasmodium, nuclei naturally divide synchronously. Thus, it offers an opportunity to study growth and mitosis within a true organism. The effects of 5 mM sodium-butyrate on these processes have been examined. When this material is added to the culture medium during mitosis, the butyrate acts like a fixative on condensed chromosomes. During interphase, this short fatty acid stops growth and immediately inhibits DNA synthesis. Furthermore, it prevents differentiation in macroplasmodia induced to spherulate. All these modifications are readily reversible after transfer to a medium lacking butyrate. PMID- 7471172 TI - Hormonal modulation of gap junctions in rat ovarian follicles. AB - Homocellular gap junctions between granulosa cells and between theca interna cells, and heterocellular gap junctions between granulosa cells and oocytes persist in rat ovarian follicles for as long as 90 days following hypophysectomy. Gonadotrophic and/or steroid hormones are therefore not required for the maintenance of gap junctions between these cells during early follicular growth. However, replacement therapy with estrogen and human chorionic gonadotrophin results in amplification of gap junctions in granulosa and theca interna cells respectively. Within 24h following hormonal stimulation, growth of gap junctions is characterized by the appearance of formation plaques as observed in freeze fracture replicas and by the association of microfilamentous material located subadjacent to gap junction membrane observable in thin-sectioned cells. PMID- 7471174 TI - Specific effect of calcium ions on the calcium-sensitive cells of the pars intermedia in the goldfish. AB - Cytological changes in the calcium-sensitive (Ca-s) cells (formerly termed PAS positive cells) of the pars intermedia were investigated in the goldfish after adaptation to deionized water (DW), with or without addition of sodium, potassium and magnesium. These ions were added as chloride salts at concentrations similar to those present in fresh water (FW). The marked stimulation of the Ca-s cells is not inhibited in DW supplemented with Na+ (0.35 mM/l), K+ (0.05 mM/l), and Mg2+ (0.2 mM/l) for a period of 24 days. The inhibition of the response to DW with calcium chloride (2 mM/l). These data show that chloride ions are not responsible for the regression of the Ca-s cells observed in goldfish kept in DW supplemented with calcium chloride. The effect of calcium ions on the Ca-s cells appears to be specific. These results support the hypothesis that the Ca-s cells synthesize a factor (hypercalcin?) involved in calcium regulation, and that its release is influenced by the calcium content of the environment. The role of the pars intermedia in calcium metabolism is strengthened by the present results. Biochemical data suggest the presence of a hypercalcemic factor in the pituitary of fish (Parsons et al. 1978) and are in agreement with the present cytological findings. PMID- 7471175 TI - The ultrastructure of the heart of Oniscus asellus L. and Asellus aquaticus L (Crustacea, Isopoda). AB - The endocardium of Oniscus asellus L. and Asellus aquaticus L. consists of lipid cells. The epicardium consists of a layer of cells with a vesiculated cytoplasm covered by a thick extracellular fibrous sheet. The myocardium is a single layer of cells, the sarcolemma invaginates at Z disc level forming transverse tubules, and longitudinal tubules branch off from these. At the A-I level longitudinal tubules from transverse systems, which form couplings with the sarcoplasmic reticulum. The sarcoplasmic reticulum appears as perforated sheets enveloping the myofibrils. Two types of nerve terminal are found: one is embedded in a myocardial cell process, the other lies in a myocardial cell depression. They contain clear and dense-cored synaptic vesicles. PMID- 7471176 TI - Zymogen granules of pancreas decrease in size in response to feeding. AB - In the first descriptions of pancreatic enzyme secretion about 100 years ago, it was noticed that zymogen granules became smaller and disappeared from the apical region of acinar cells after feeding. We have repeated these experiments and characterized changes in granule size by quantitative electron microscopy 90 min after feeding previously fasted rats. In fasted animals, granules occupied the apical portion of the cell, had an average number of 45 +/- 3 granules per cell section (+/- SE), and measured 0.85 +/- 0.15 micrometers in diameter (+/- SD). After feeding, the number and size of granules decreased. Individual samples showed either a decrease in size alone or a decrease in both size and number, but in no case did they show a reduction in granule number alone. The mean diameter of granules decreased to 0.65+/- 0.15 microns (+/- SD) or about a 55% reduction in average granules volume as compared to controls (0.32 vs. 0.14 microns 3). The size distributions were unimodal and normal in both fasted and fed rats; however, in fed animals, the distribution was shifted to lower values (diameter range 0.40 1.40 microns for fasted rats vs. 0.10-1.30 microns for fed rats). The number of granules decreased to an average of 29 +/- 2 granules per cell section (+/- SE) after feeding, and, on the average, samples with the most granules had larger ones than samples with the fewest granules. The present results support the original observations on live rabbit pancreas that individual granules decrease in size in response to feeding. We suggest that these size changes reflect the loss of proteins across the granule membrane as proposed by the equilibrium hypothesis for digestive enzyme secretion. PMID- 7471177 TI - Fine structural study of the statocysts in the veliger larva of the nudibranch, Rostanga pulchra. AB - The two statocysts of the veliger larva of Rostanga pulchra are positioned within the base of the foot. They are spherical, fluid-filled capsules that contain a large, calcareous statolith and several smaller concretions. The epithelium of the statocyst is composed of 10 ciliated sensory cells (hair cells) and 11 accessory cells. The latter group stains darkly and includes 2 microvillous cells, 7 supporting cells, and 2 glial cells. The hair cells stain lightly and each gives rise to an axon; two types can be distinguished. The first type, in which a minimum of 3 cilia are randomly positioned on the apical cell membrane, is restricted to the upper portion of the statocyst. The second type, in which 9 to 11 cilia are arranged in a slightly curved row, is found exclusively around the base of the statocyst. Each statocyst is connected dorso-laterally to the ipsilateral cerebral ganglion by a short static nerve, formed by axons arising from the hair cells. Ganglionic neurons synapse with these axons as the static nerve enters the cerebral ganglion. The lumen of the statocyst is continuous with a blind, constricted canal located beneath the static nerve. A diagram showing the structure of the statocyst and its association with the nervous system is presented. Possible functions of the statocyst in relation to larval behavior are discussed. PMID- 7471178 TI - A retinal projection to the lateral hypothalamus in the rat. AB - This study presents evidence for a retinal projection to neurons in the lateral hypothalamic area (LHA) of the albino rat. In Golgi-Kopsch material dendrites from LHA-neurons are observed to extend through the supraoptic commissures into the optic tract. The presence of dendrites in the optic tract is confirmed by electron microscopy. Numerous axon terminals are observed forming asymmetric synaptic contacts with these dendritic profiles. Following bilateral enucleation, many of the preterminal axons and terminals in synaptic contact with dendrites in the optic tract demonstrate dark degeneration. After intraocular injection of horseradish peroxidase, there is marked labeling of preterminal axons and terminals in the optic tract. These observations indicate that LHA neurons receive a direct retinal projection from terminals making synaptic contact with dendrites of LHA-neurons extending into the optic tract. PMID- 7471179 TI - Development of blood-cerebrospinal fluid barrier to horseradish peroxidase in the avian choroidal epithelium. AB - The penetration of horseradish peroxidase (HRP) from blood into ventricle via the avian choroid plexus was examined by electron microscopy in order to study the development of the blood-cerebrospinal fluid barrier (BCSFB). 1-20 mg HRP was injected into the allantoic vein of chick embryos between the 7th and 21st day of incubation. Until the 8th day (stage 34) the reaction products of injected HRP were observed in the interepithelial clefts at both luminal (ventricular) and abluminal sides. At the 9th day (stage 35), their penetration was blocked at most apical junctional complexes of the choroidal epithelia. At the 10th day (stage 36) and at every subsequent stage, HRP molecules were completely impeded at the apical tight junctions. These results show that the BCSFB to HRP in the avian choroidal epithelium is completely established by the 10th day, when molecules of HRP are still able to permeate between the capillary endothelia in the cerebellum and the spinal cord (Wakai and Hirokawa 1978). PMID- 7471180 TI - The effect of l-thyroxine on the metamorphosis of Ascidia malaca. AB - Larvae of Ascidia malaca, both before and after hatching, were treated with L thyroxine solutions. The effect of the thyroid hormone was to induce the onset of metamorphosis and then to cause the rate at which body reorganization occurred to increase. In treated larvae the resorption of the tail occurred only few hours after hatching, and a beating heart appeared form 10 to 15 h earlier than in the control larvae. These results are discussed in the context of a probable relationship between the occurrence of a hormonal metamorphic factor and the button cells of the trunk. PMID- 7471181 TI - The temporo-spatial course of degeneration after cutting cortico-cortical connections in adult rats. AB - Adult albino rats received callosotomies or lesions in the paracingular cortex. Between 12 h and 3 months after injury the structure and topography of the degeneration products were studied by light- and electron-microscopy. The degeneration process was quantified by television-image analysis applied to sections prepared according to a new technique that stains reliably degenerating terminals and lysosomes (Gallyas et al. 1980). All types of cortico-cortical connections show a multiphasic degeneration process: During a precursor stage a small number of dense bodies and mitochondrial granules are stained. These and the few early degenerating axon terminals are much more diffusely distributed than the large number of terminals that degenerate during the following period. The terminal degeneration shows a biphasic time course. One maximum appears at 2 7 days post operation, which corresponds to the well known direct consequence of axotomy. The second peak at 10-20 days post operation could be caused by transneuronal reorganization of the cortical connectivity. Terminal degeneration always begins along the borders between cortical regions and areas, but it may change its laminar and columnar distribution pattern during the second phase. The degeneration products that are phagocytosed by astrocytes seem to be removed by intracellular transport to their perivascular endfeet. The degeneration process ends with fiber degeneration which, especially in laminae I and VI, may form a separate peak after 20 days or more. PMID- 7471182 TI - Evidence of membrane transformation during melanogenesis. Electron microscopic study on the retinal pigment epithelium of chick embryos. AB - Some characteristics of early premelanosomes (PM) suggest that primarily a continuous cisternal complex of the endoplasmic reticulum (ER) is transformed simultaneously to PM. These characteristics are: (i) the form and size, which are similar to ER cisternae; (ii) the localization in groups in the ER; )iii) the same stage of maturation within a group; (iv) the continuities between early PM, and (v) the lack of continuities between ER and PM. Comparative measurements reveal that the limiting membrane of PM, with a total thickness of 7.6 +/- 0.19 nm and a center-to-center distance of 5.2 +/- 0.06 nm, is significantly thicker than the ER membrane (6.3 +/- 0.15 nm and 4.3 +/- 0.04 nm, respectively) and the melanosome limiting membrane (6.5 +/- 0.22 nm and 4.4 +/- 0.05 nm, respectively). Therefore, during the formation of melanosomes, the limiting membrane must be transformed from a thin (ER) to a thick (PM) and again to a thin (melanosome)stage. PMID- 7471183 TI - Freeze-fracture and deep-etching studies on zymogen-granule membranes of the rat pancreas. AB - Whole pancreatic zymogen granules or their membrane fraction were examined by freeze-fracture or deep-etching under different experimental conditions. The granules were fixed for different time periods, or not fixed, and were cryoprotected with glycerol or DMSO; 3% glutaraldehyde followed by 30% glycerol were finally chosen for giving the best resolution and the highest density of intramembrane particles (IMP). IMP are present on the PF and EF leaflets. Their number decreases with the duration of the fixation. Several granules exhibit IMP free blebs. Incubation of the granules with protamine sulfate causes an aggregation of IMP and of the rough-textured background on the EF leaflet. A second fracture plane can be formed and has been shown by deep-etching to be intercalated between PF and EF. Deep-etching has also shown that particles attached to the perimeter of the granules and of the blebs are, in fact, large nodules on the PS face which partially extend onto the blebs and do not aggregate with the IMP after protamine treatment. Fusion is also indicated between membrane vesicles. Freeze-fracture of the purified membrane fraction seems to indicate the formation of an IMP cap during the lysis of the granule. Moreover, large nodules remain present on the PS face on these membrane fractions but the majority disappear after washing at pH 11.2 with Na2CO3 and EDTA. PMID- 7471184 TI - Quantitative analysis of muscle breakdown during starvation in the marine flatfish Pleuronectes platessa. AB - The present study describes the effects of starvation for a duration of four months on the ultrastructure of skeletal muscles from the marine flatfish (Pleuronectes platessa L.). Starvation is associated with a decrease in resting metabolic rate from 20.1 +/- 2.2 to 11.6 +/- 1.5 mg . O2/kg/h (P less than 0.05) and muscle wasting. Median fibre size fell from 700 micrometer 2 to 500 micrometer 2 in intermediate (fast oxidative) and from 1,800 micrometer 2 to 600 micrometer 2 in starved, white (fast-glycolytic) muscle fibres. In contrast, median fibre size in red (slow oxidative) muscle remained within the range 300 400 micrometer 2. The fraction of red fibre volume occupied by myofibrils (58.6%) and mitochondria (24.5%) did not change significantly with starvation. There was, however, a decrease in stored lipid 110.7% to 3.2%) and an alteration in the structure of the cristae in mitochondria from red muscle. Atrophy of white muscle fibres is associated with a decrease in both the diameter and fractional volume occupied by myofibrils (85.7% to 61.9% P less than 0.01). In a high proportion of white fibres peripheral degeneration of Z-discs is evident causing an unravelling of the thin filament lattice. It is suggested that this allows a partial decrease in myofibril diameter and hence the maintenance of contractile function in muscle from starved fish. In severely degenerating white fibres, disorganised thick and thin filaments and numerous multi-membrane lysosome-like vesicles are observed. Starvation results in an increase in the average content of mitochondria in white fibres from 2.2 to 6.7% (P less than 0.01). In fed plaice mitochondria constitute less than 1% of the volume of the white fibre in 43.5% of the fibres. The proportion of white fibres containing more than 6% mitochondria increases from 6.5% to 58% with starvation. PMID- 7471185 TI - Hypothalamic lesions stimulating growth hormone cell activity in the goldfish. AB - The cytology of the growth-hormone (GH) cells of the goldfish pituitary were examined following electrothermic lesions of the anterior preoptic hypothalamus and telencephalon. Following lesions of the nucleus preopticus (NPO) light microscopy of the pituitary revealed a significant increase in the nuclear diameter and a degranulation of the GH cells. Lesions of the telencephalon anterior or dorsal to the NPO had no cytological effect on the GH cells. The ultrastructural appearance of the GH cells of NPO-lesioned fish was characterized by a marked degranulation of the cytoplasm and a proliferation of the rough endoplasmic reticulum indicative of enhanced secretory activity. The GH cells of the proximal pars distalis (PPD) are directly innervated by peptidergic (type A) and aminergic-like (type B) neurosecretory axons. Following lesions of the NPO, there was a marked reduction in the number of type A fibers in the PPD. These results suggest that the type A fibers innervating the GH cells originate in the NPO and act to inhibit the secretory activity of the GH cells. PMID- 7471186 TI - Ultrastructural localization of endogenous peroxidase in the lower respiratory tract of the guinea pig. AB - Endogenous peroxidase activity was demonstrated by cytochemistry in mucous cells of the submucosal glands and tracheobronchial epithelium of guinea pigs. It is localized in the nuclear envelope, in cisternae of rough endoplasmic reticulum, and in secretory granules. It was not seen in Golgi saccules or in the airway lumen. By contrast, all epithelial cells within the lung including mucous (goblet) cells lack activity. Reaction product is also absent from alveolar macrophages and mast cells. The appearance of peroxidase in mucous cells is age related. No activity was seen at 1.5 ms of age. A few mucous cells were positive at 2.5 and 3 ms while the proportion of positive cells increased substantially up to 7 ms. Thus, the age of guinea pigs in HRP transport studies must receive careful consideration in order to avoid misinterpretation of results. The function of mucous cell peroxidase is unknown. The results of this study suggest that it is secreted. Whether it plays a significant role in lung defense through its well documented anti-infectious properties remains to be determined. PMID- 7471187 TI - The origin of pituitary cysts in the rostral pars distalis of the nine-spined stickleback, Pungitius pungitius L. AB - Pituitary cysts in the nine-spined stickleback, Pungitius pungitius L., developed next to blood vessels passing through the prolactin zone of the rostral pars distalis to the connective tissue capsule at its periphery. Cysts were most frequent when pituitaries were large compared with body lengths. However, the incidence of cysts could not be closely related to body length alone. As the rostral pars distalis is more highly vascularised in fish with large pituitaries, and as there was no evidence of accumulating secretion around these blood vessels, it is suggested that cysts develop when vascular demands (or supplies) become excessive. Associated with the greater vascularity of the rostral pars distalis in large pituitaries was a remarkable development of non-granulated cells. Indeed cysts may originate as enlarged intercellular spaces between such cells, as the latter often surround blood vessels. The non-granulated cells are also important in enlarging cyst cavities (by phagocytosing prolactin-cell debris) and perhaps for maintaining their structural integrity. It is suggested that the macrophages within cyst cavities are derived from the non-granulated cells. PMID- 7471188 TI - College of General Practice 1980 Robbie Gibson Memorial Lecture. "Doctors--health promoters or sickness professionals?". PMID- 7471190 TI - Adverse drug reactions in a developing country: problems encountered in a surveillance programme in a Nigerian hospital. PMID- 7471189 TI - The histocompatibility (HLA) antigen distribution in multiple sclerosis patients in Zimbabwe. PMID- 7471191 TI - Dermatological aphorisms contact dermatitis. PMID- 7471192 TI - A symposium on hydatidosis. PMID- 7471193 TI - A mission hospital in the eastern highlands of Zimbabwe. PMID- 7471194 TI - Unusual combination of locked twins. PMID- 7471195 TI - Cimex lectularis as a vector of Hepatitis B. PMID- 7471196 TI - Dermatofibrosarcoma protuberans. PMID- 7471197 TI - A mission hospital in India. PMID- 7471198 TI - Myocardial bridges: a new horizon in the evaluation of ischemic heart disease. PMID- 7471199 TI - The diagnostic accuracy of combined clinical and noninvasive cardiac evaluation: comparison with findings at cardiac catheterization. AB - The accuracy of combined clinical and noninvasive cardiac diagnostic evaluation was prospectively examined in 108 consecutive patients referred to the heart station for echocardiographic examination prior to cardiac catheterization. History, physical examination, scalar electrocardiology, chest roentgenography, phonocardiography and pulse recording, and M-mode echocardiography were employed by the heart station cardiologist, who assigned one or more diagnoses to each patient. In addition, one of three management strategies was proposed for each patient: 1) surgery without cardiac catheterization; 2) medical therapy without cardiac catheterization; or 3) cardiac catheterization for clarification of the diagnosis. The results of the combined clinical and noninvasive evaluation were independently reviewed for each patient and compared with the diagnosis determined by cardiac catheterization, results of cardiac surgery, and total hospital course. Diagnostic predictions employing combined clinical and noninvasive cardiac evaluation were completely correct in 86% of patients, and management strategy was correct in 97% of individuals. In approximately one-half of all patients full cardiac catheterization or coronary arteriography was recommended. All management strategy errors and two-thirds of diagnostic errors occurred in patients with mitral regurgitation, aortic regurgitation, or coronary artery disease. Combined clinical and noninvasive evaluation results in accurate diagnostic information adequate for the formulation of appropriate management strategies in the majority of patients, but many individuals with cardiac disease still require invasive evaluation for complete diagnosis. PMID- 7471200 TI - To invade or not to invade. PMID- 7471201 TI - An improved snare system for the nonsurgical retrieval of intravascular foreign bodies. AB - We have modified a snare technique originally described for transcolonoscopic removal of colonic polyps for the retrieval of intravascular foreign bodies. The key difference from other snare techniques is the employment of a wire snare with a crimp in its midportion that enables the formation of a loop in a plane perpendicular to the axial direction of the guiding catheter. With this technique we have quickly removed a variety of objects from the right heart and pulmonary arteries in four patients. PMID- 7471202 TI - Refractory variant angina controlled with combined drug therapy in a patient with a single coronary artery. AB - A young man with a single left coronary artery and refractory variant angina is described. Spontaneous coronary artery spasm developed during coronary arteriography at the site of a 50% fixed left anterior descending coronary artery stenosis. Frequent episodes of rest angina with transient ST segment elevation persisted in hospital in spite of treatment with three different calcium antagonist drugs. Symptoms disappeared only when the combination of nifedipine, diltiazem, isosorbide dinitrate, and nitroglycerin ointment were given. Ergonovine testing was used to objectively assess the response to treatment. PMID- 7471204 TI - Cardiac dimensional analysis by use of biplane cineradiography: description and validation of method. AB - Biplane cineradiography is a potentially powerful tool for precise measurement of intracardiac dimensions. The most systematic approach to these measurements is the creation of a three-dimensional coordinate system within the x-ray field. Using this system, interpoint distances, such as between radiopaque clips or coronary artery bifurcations, can be calculated by use of the Pythagoras theorem. Alternatively, calibration factors can be calculated in order to determine the absolute dimensions of a structure, such as a ventricle or coronary artery. However, cineradiography has two problems that have precluded widespread use of the system. These problems are pincushion distortion and variable image magnification. In this paper, methodology to quantitate and compensate for these variables is presented. The method uses radiopaque beads permanently mounted in the x-ray field. The position of the bead images on the x-ray film determine the compensation factors. Using this system, measurements are made with a standard deviation of approximately 1% of the true value. PMID- 7471205 TI - A simple device to reproduce the spatial orientations of the thorax and transducer for serial M-mode and two-dimensional echocardiograms. AB - We have described a device consisting of three circular levels for locating the spatial orientations of the thorax and a transducer that can be used to reproduce the path of the sonic beam through the heart. PMID- 7471203 TI - Rationale for ergonovine testing on the CCU. PMID- 7471206 TI - The use of a microcomputer in the evaluation of echocardiograms. AB - A low-cost microcomputer system is used to calculate cardiac parameters from echocardiographic (Echo) measurements and produce a chartable report. The system is simple, uses Basic language, and provides for data storage. PMID- 7471207 TI - The spontaneous insertion of proteins into and across membranes: the helical hairpin hypothesis. AB - We propose that the initial event in the secretion of proteins across membranes and their insertion into membranes is the spontaneous penetration of the hydrophobic portion of the bilayer by a helical hairpin. Energetic considerations of polypeptide structures in a nonpolar, lipid environment compared with an aqueous environment suggest that only alpha and 3(10) helices will be observed in the hydrophobic interior of membranes. Insertion of a polypeptide is accomplished by a hairpin structure composed of two helices, which will partition into membranes if the free energy arising from burying hydrophobic helical surfaces exceeds the free energy "cost" of burying potentially charged and hydrogen bonding groups. We suggest, for example, that the hydrophobic leader peptide found in secreted proteins and in many membrane proteins forms one of these helices and is oriented in the membrane with its N terminus inside. In secreted proteins, the leader functions by pulling polar portions of a protein into the membrane as the second helix of the hairpin. The occurrence of all categories of membrane proteins can be rationalized by the hydrophobic or hydrophilic character of the two helices of the inserted hairpin and, for some integral membrane proteins, by events in which a single terminal helix is inserted. We propose that, because of the distribution of polar and nonpolar sequences in the polypeptide sequence, secretion and the insertion of membrane proteins are spontaneous processes that do not require the participation of additional specific membrane receptors or transport proteins. PMID- 7471208 TI - Single extrachromosomal ribosomal RNA gene copies are synthesized during amplification of the rDNA in Tetrahymena. AB - A novel form of extrachromosomal rDNA has been identified in conjugating Tetrahymena cells. This rDNA consists of 11 kb linear double-stranded DNA molecules, each containing a single rRNA gene copy. The DNA sequence, tandemly repeated CCCCAA (Blackburn and Gall, 1978) found at the termini of extrachromosomal palindromic rDNA (the macronuclear form found in vegetatively growing cells), is also present at the corresponding terminus of the 11 kb rDNA. The other end of this molecule has an extra 0.3 kb segment of DNA covalently attached to the DNA region corresponding to the center of the palindromic rDNA. The kinetics of appearance and synthesis of the 11 kb rDNA early in macronuclear development are consistent with its being an intermediate in rDNA amplification. PMID- 7471209 TI - Adenovirus early gene products may control viral mRNA accumulation and translation in vivo. AB - The mechanisms controlling early adenovirus gene expression in vivo have been studied using inhibitors of protein synthesis. When inhibitors were added shortly before or at the onset of infection, viral mRNA from all early regions was transcribed, spliced and accumulated over a 7 hr period. After longer pretreatment, accumulation of several early mRNAs were suppressed. Addition of inhibitors 1 hr after infection enhanced the accumulation of viral mRNA in the cytoplasm. Translation of early mRNA selected on adenovirus DNA in a cell-free system reflected the amount of viral mRNA present. A viral coded product may therefore control accumulation of viral mRNA. A different pattern emerged when inhibitors of protein synthesis were removed at 5 hr postinfection and cells were removed at 5 hr postinfection and cells were pulse-labeled in vivo. If inhibitors were introduced at or before infection, early viral proteins were synthesized only after a lag of 1-3 hr. However, if treatment was introduced 1 hr postinfection, reversion of the protein synthesis block was instantaneous. It appears that protein synthesis inhibitors reveal an in vivo translational block for viral mRNA. This block could be overcome by preinfection with a related virus. Furthermore, no block was observed in a virus-transformed human embryonic kidney cell line (293) which expresses early region 1 of the viral genome. Viral gene product(s) encoded in early region 1 may control translation of early adenovirus messenger RNA in vivo. PMID- 7471210 TI - Identification of the gene and mRNA for the adenovirus terminal protein precursor. AB - The precursor of the 55K adenovirus terminal protein is an 87K protein that is covalently linked to viral DNA. This protein is likely to be identical to the 80,000 dalton protein described by Challberg et al. (1980). The mRNA for the 87K terminal protein precursor, like that for the E2-72K DNA binding protein, is detectable at both early and late times of infection, and its production is sensitive to protein synthesis inhibition (Lewis and Mathews, 1980). The 87K protein, together with proteins of 105,000 and 75,000 daltons, are translated from leftward transcribed (1-strand) messenger RNAs that are complementary to the viral genome between positions 11.2 and 31.5. Additional hybridization to the region between coordinates 37.3 and 41 suggests that the RNA body is spliced to sequences mapping farther right in the genome. Electron microscopic heteroduplex analysis has revealed a family of 1-strand RNAs that probably encode these proteins. The RNA bodies extend from coordinated 30, 26 and 23 to 11.1, with leaders at 39, 68.5 and 75 map units, defining a new adenovirus early region. These RNAs and region E2 RNAs share the first leader and presumably the same promoter, and may be coordinately expressed. Virions of the protease-deficient adenovirus 2 mutant ts1 grown at the restrictive temperature contain only the 87K form; when grown at the permissive temperature they contain both the 87K and 55K forms, and an additional 62K form; wild-type virions contain only the 55K form. Peptide analysis shows all these proteins to be related. The DNA-protein complex containing the 87K form is active as a template for viral DNA replication in vitro. This data supports a model of adenovirus DNA replication in which the 87K terminal protein precursor is the primary translation product and primes DNA synthesis. The 87K precursor is processed curing virus maturation to the 55K terminal protein, possibly via a 62K intermediate form, by the virus-specified Ad2ts1 protease. PMID- 7471211 TI - The synthesis and deployment of filamin in chicken skeletal muscle. PMID- 7471212 TI - Studies on the galactose-binding site of ricin and the hybrid toxin Man6P-ricin. AB - N-acetylimidazole (NAI) was used to O-acetylate the plant seed toxin ricin. O acetylation of one to two tyrosine residues per molecule of ricin inhibited ricin binding to Sepharose 4B and decreased toxicity by 90% in a protein synthesis inhibition assay in HeLa cells. Lactose, known to block the binding site on the ricin B subunit, protected ricin from NAI modification of binding or toxicity. Thus NAI, under these conditions, can be a lactose site-specific inhibitor. The lactose site-specific modification of the hybrid toxin, Man6P-ricin, performed under the same conditions, exhibited the same 90% inhibition of Man6P receptor mediated toxicity as the galactose-containing receptor-mediated toxicity of either Man6P-ricin or ricin. Thus the ricin B chain lactose-binding site appears to be essential for the high potency of Man6P-ricin via the new cell type specific Man6P receptor. Treatment of fibroblasts with neuraminidase exposes galactose residues, thus increasing the sensitivity to ricin eight fold. The Man6P receptor-mediated toxicity of Man6P-ricin is not affected by this treatment, although the galactose-inhibited route is potentiated eight fold. The Man6P-ricin hybrid appears to require the ricin B chain galactose-binding site to enter the cytosol after initially binding to the Man6P receptor. These data provide some insights into the proper design of hybrid toxins. We discuss a number of possible models for hybrid toxin entry. PMID- 7471213 TI - The ovalbumin gene family: hormonal control of X and Y gene transcription and mRNA accumulation. AB - The ovalbumin gene family is composed of three genes, X, Y and ovalbumin, which are expressed in laying hen oviduct. We have analyzed the in vivo transcription products of X and Y genes and the effect of steroid hormones on their synthesis and accumulation. As in the case of ovalbumin, the complete gene transcripts and processing intermediates are present in the poly(A)+ RNA fraction. The mature RNAs are found in polysomes and are translated into proteins. The expression of X and Y genes is controlled by steroid hormones: X and Y RNAs are not detectable in oviducts from chicks withdrawn from estrogen stimulation, whereas in chicks stimulated with estrogen for 7 days, X RNA represents 0.3% and Y RNA 0.8% of ovalbumin mRNA. In laying hen, however, the levels of X and Y RNAs are about 2% of ovalbumin mRNA. After stimulation with other steroid hormones, alone or in combination, the level of X and Y RNA does not achieve that detected in laying hen. Progesterone has a much weaker effect on X RNA accumulation than on that of Y and ovalbumin mRNAs. Studies with isolated nuclei show that X and Y gene expression is regulated by hormones at the level of transcription. However, the differences observed between the transcription rates and the accumulation of X and Y mRNAs suggest that the expression of X and Y genes could also be controlled at the levels of RNA processing and/or mRNA stability. PMID- 7471214 TI - A transcription map for the rabbit beta-globin gene. AB - We have characterized the transcripts of the beta-globin gene in RNA isolated from rabbit bone marrow using the S1 mapping procedure of Berk and Sharp (1977). The largest beta-globin-specific RNA that we have detected is a transcript, colinear with the gene, that contains the mRNA coding sequences plus both intervening sequences but does not extend into the extragenic DNA sequences. The 5' and 3' ends of this pre-mRNA maps at the same position as those of mature globin mRNA. Several partially spliced RNAs are described that are probably intermediates in the pathway from the colinear transcript to mature mRNA. The largest spliced pre-mRNA has lost only about 40 of the 126 nucleotides of the small intervening sequence and contains the entire large intervening sequence together with the coding sequences. A second major intermediate pre-mRNA lacks only the entire small intervening sequence. Finally, a pre-mRNA is described in which all but approximately 90 nucleotides of the large intervening sequence have been excised. Thus both intervening sequences appear to be able to be excised in a stepwise fashion. Removal of the small intervening sequence seems to precede the removal of the large intervening sequence. PMID- 7471215 TI - Phosphorylation and dephosphorylation of chromosomal proteins during in vitro transcription. PMID- 7471216 TI - Age-dependent changes in the capacity of transdifferentiation of retinal pigment cells as revealed in clonal cell culture. PMID- 7471217 TI - Analysis of developmentally homogeneous neural crest cell populations in vitro. III. Role of culture environment in cluster formation and differentiation. PMID- 7471218 TI - Surface proteins on Cloudman melanoma cells. PMID- 7471219 TI - BCG-induced macrophage suppression in mice: suppression of specific and nonspecific antibody-medicated and cellular immunologic responses. PMID- 7471221 TI - [Worldwide aspects of teaching the subject, "dosage forms"]. PMID- 7471220 TI - Loss of target-cell plasma-membrane protein induced by sensitized allogeneic lymphocytes. PMID- 7471222 TI - [Antitubercular agents. XXII. Physical and physico-chemical properties of thiobenzamides. III. 13C-NMR spectra]. PMID- 7471223 TI - [Synthesis of amino alcohols with potential bronchodilator effects]. PMID- 7471224 TI - [Changing the solubilizing capacity of sodium taurocholate and selected tensides using electrolytes]. PMID- 7471225 TI - [General principles of the effects of thyroid hormones]. PMID- 7471226 TI - [Use of biosynthetic materials in animal feed and veterinary practice]. PMID- 7471227 TI - [Carrier isotachophoresis and its potential in pharmaceutical analysis. I. Anion analysis]. PMID- 7471228 TI - [Detection of pharmaceutical aids in drugs and cosmetic preparations using thin layer chromatography]. PMID- 7471229 TI - [Colorimetric determination of chenodeoxycholic acid after elution from thin layer Silufol]. PMID- 7471230 TI - [Practical use of rheology in quality testing of dosage forms]. PMID- 7471231 TI - [Comparison of the local anesthetic activity of the eye drops Novesine and Alcain in laboratory experiments]. PMID- 7471232 TI - [Endoscopic study of the esophagus in esophageal carcinoma associated with carcinoma in the otorhinolaryngeal area (indications, technics and results)]. PMID- 7471233 TI - [External pancreatic secretion after temporary obstruction of the duct, venostasis and ischaemia of the pancreas in the dog (author's transl)]. PMID- 7471234 TI - [Histochemistry of the human duodenal mucous membrane with special emphasis on the proximo-distal gradient of enzymatic activities in the enterocyte brush border]. PMID- 7471235 TI - [Lambliasis in duodenobiliary region (author's transl)]. PMID- 7471236 TI - [Contribution of ultrasonics with the gray scale to the diagnostis of hepatobiliary diseases (author's transl)]. PMID- 7471237 TI - [Development of hepatology in the Democratic Republic of Germany]. PMID- 7471238 TI - [Chronic functional gastric dyspepsia followed up over years (author's transl)]. PMID- 7471239 TI - [The excellence of scientific-medical information - a prerequisite of excellence in the scientific professions]. PMID- 7471240 TI - [Informative values of sequential test expressing adrenal and ovarian suppression in the differential diagnosis of hirsutism complicated by menstrual dysfunction (author's transl)]. PMID- 7471241 TI - [The use of a new Czechoslovak preparation "Anecotan" in obstetrical analgesia (author's transl)]. PMID- 7471242 TI - [Intrauterine growth retardation from the aspect of prenatal care]. PMID- 7471243 TI - [Organic components in amniotic fluid in relation to fetal dystrophy]. PMID- 7471244 TI - [Fibrolipoleiomyoma of the uterine cervix]. PMID- 7471245 TI - [Lactation (pathophysiology and clinical aspects)]. PMID- 7471246 TI - [Acupuncture and its possibilities in gynecology and obstetrics]. PMID- 7471247 TI - [Perspectives in the use of the theory of reliability in gerontology]. PMID- 7471248 TI - [Experimental model of proliferative pseudoretinopathy]. PMID- 7471249 TI - [Ablatio centralis serosa - experience with photocoagulation treatment (author's transl)]. PMID- 7471250 TI - [Treatment of vascular retinal lesions with Cavinton (author's transl)]. PMID- 7471251 TI - [Treatment of diabetic retinopathy with photocoagulation]. PMID- 7471252 TI - [Treatment of occlusion of central retinal vein branches using photocoagulation. A preliminary report]. PMID- 7471253 TI - [Bilirubin content in the subretinal fluid and its dependence on the duration of the retinal detachment (author's transl)]. PMID- 7471254 TI - [Solar retinitis (author's transl)]. PMID- 7471255 TI - [Two cases of rate vascular retinal diseases in young soldiers (author's transl)]. PMID- 7471256 TI - [Possibilities of ocular lesions in work with lasers (author's transl)]. PMID- 7471257 TI - [Results of conservative treatment of strabismus in 778 children during last 15 years, treated in ambulatory conditions (author's transl)]. PMID- 7471258 TI - [Ultrastructure of middle ear cholesteatoma]. PMID- 7471259 TI - [Tympanometry after tympanoplasty operations]. PMID- 7471260 TI - [Evaluation of hearing after kophosurgery for hearing disorders. II. The importance of the Sullivan and Federici-Lewis tests in hearing loss operations]. PMID- 7471261 TI - [Wound healing after laryngectomy]. PMID- 7471262 TI - [Speech intelligibility after laryngectomy]. PMID- 7471263 TI - [The oncology program of the otolaryngology department of the regional health system in Banska Bystrica]. PMID- 7471264 TI - [Stenoses of the larynx and upper trachea]. PMID- 7471265 TI - [Mucus producing elements and mucus production in the paranasal sinuses]. PMID- 7471266 TI - [Pneumosinus frontalis dilatans]. PMID- 7471267 TI - [Radiographic examination in the diagnosis of inhaled foreign bodies]. PMID- 7471268 TI - [Ear injuries caused by lightning]. PMID- 7471269 TI - [Chylothorax as a result of a stab injury of the neck]. PMID- 7471270 TI - [A rotation trephine for a safe, bilateral separation of the cochlea]. PMID- 7471271 TI - [Spinal cord symptomology in diseases of the descending aorta]. PMID- 7471272 TI - [Experimental echoencephalography]. PMID- 7471273 TI - [Neurologic manifestations of chronic alcoholism]. PMID- 7471274 TI - [Analysis of fatty acids in lecithins and cholesterol esters in the distal and proximal portions of lumbar cerebrospinal fluid]. PMID- 7471275 TI - [Neurological findings in mentally handicapped children and adolescents in the institute in Budenicky. Part IV]. PMID- 7471276 TI - [Share of the influence of the visible part of solar radiation in the development of psychoses with disorders of affectivity (author's transl)]. PMID- 7471277 TI - [Results of treatment by Moditen Depot in outpatients' wards in the South Moravian Region (author's transl)]. PMID- 7471278 TI - [Conditioned-reflex therapy of enuresis by means of the device ROE-70 (author's transl)]. PMID- 7471279 TI - [A case of acute agranulocytosis during treatment by clozapine (author's transl)]. PMID- 7471280 TI - [Experience with group psychotherapy in the conditions of district and works psychiatric outpatients' care (author's transl)]. PMID- 7471281 TI - [CPRS - new psychopathological scale of Scandinavian and English authors (author's transl)]. PMID- 7471282 TI - [Children systematically damaging clothes and furnishings (author's transl)]. PMID- 7471283 TI - [Diabetes mellitus in childhood]. PMID- 7471284 TI - [Delayed pubertal development in pediatric practice]. PMID- 7471285 TI - [Physiological requisites for successful lactation]. PMID- 7471286 TI - [New projections for the population of Czechoslovakia up to the year 2000]. PMID- 7471287 TI - [Foster family care from the viewpoint of the pediatrician]. PMID- 7471288 TI - [Treatment of Hodgkin's disease in children]. PMID- 7471289 TI - [Cephalocele]. PMID- 7471290 TI - [Congenital adrenal hypoplasia (author's transl)]. PMID- 7471291 TI - [Influenza vaccination of children with a sub-unit vaccine made in Czechoslovakia]. PMID- 7471292 TI - [Probability of infant death by age groups in Czechoslovakia and Slovakia from 1966-1978]. PMID- 7471293 TI - [Unconvincing results of repeated intranasal vaccination of school children against influenza]. PMID- 7471294 TI - [Cerebral ischemia in children and adolescents]. PMID- 7471295 TI - [Mitral valve prolapse syndrome in children and adolescents]. PMID- 7471296 TI - [Ultrastructure of the liver in asphyctic newborns]. PMID- 7471297 TI - [Levels of riboflavin in the blood of normal newborns and during phototherapy for hyperbilirubinemia]. PMID- 7471298 TI - [Diagnostic problems in infants with asymmetrical gonadal dysgenesis]. PMID- 7471299 TI - [Changes in immunoreactivity in children with recurrent respiratory infections]. PMID- 7471301 TI - [Chronic recurrent parotitis]. PMID- 7471300 TI - [Pathogenic aspects of the hemolytic-uremic syndrome. (The toxic effect of neuraminidase)]. PMID- 7471302 TI - [Immunologic activity of professional phagocytes (granulocyte and monocyte macrophages) in newborns]. PMID- 7471303 TI - [Social medicine and its relation to the organization of health services]. PMID- 7471304 TI - [Analytical approach to the study of differences in operational per capita costs in hospitals administered by National Committees in various regions of Czechoslovakia]. PMID- 7471305 TI - [Care of the aged in England]. PMID- 7471306 TI - [Use of computer technics in the management of epidemic control]. PMID- 7471307 TI - [An international scientific and medical information system in socialist countries]. PMID- 7471308 TI - [Medico-geographic spread of certain diseases in Slovakia in relation to gerontological problems]. PMID- 7471309 TI - [From a working group of social hygienists to the Society of Social Hygiene of the Democratic Republic of Germany]. PMID- 7471310 TI - Mixed-function oxidase enzymes in adriamycin-sensitive and resistant sublines of P-388 leukemia. AB - Levels of mixed-function oxidase (MFO) enzymes were measured in adriamycin(ADR) sensitive murine leukemia P-388 and its ADR-resistant subline P-388/ADR. The subcellular fractions of the resistant cells showed decreased contents of MFO components, cytochrome P-450 and cytochrome b5, in comparison with the identically prepared fractions of the parental tumor. Similarly, the levels of 7 ethoxycoumarin O-deethylase and the rate of ascorbate induced lipid peroxidation in vitro showed lower values in resistant tumor cells than those of P-388 tumor cells. The observed differences in the two tumor cell types were found to be considerably enhanced if the tumor cells were exposed in vitro to ADR before fractionation. The magnitude of induction of the MFO enzymes was significantly greater in the ADR exposed P-388 cells. The corresponding inducibility was suppressed in the drug exposed resistant tumor cells. PMID- 7471311 TI - Carcinogen-DNA interaction: complexing of distamycin-A with DNA is necessary for its inhibitory effect on the N-methyl-N-nitrosourea induced methylation of DNA. PMID- 7471312 TI - Effect of 3-methylcholanthrene administration on hepatic ribonucleic acid polymerase activities. AB - The effect of the in vivo administration of 3-methylcholanthrene upon rat hepatic RNA polymerase activities was investigated. Aggregate RNA polymerase activity assayed in liver nuclei was stimulated by 33% over control. Characterization of the individual RNA polymerase activities by virtue of their differential sensitivity to alpha-amanitin revealed that RNA polymerase I activity was maximally increased by 70% at approx. 16 h post-administration of the polycyclic hydrocarbon; RNA polymerase II activity was stimulated by 33%. The kinetics of RNA polymerases I and II stimulation differed in that the nucleolar enzyme's activity increased earlier and peaked later. RNA polymerase III activity was not significantly different from control. Phenobarbital, another inducer of the mixed function oxidases, had essentially no effect on the activity of hepatic RNA polymerases. Solubilization of the RNA polymerases followed by separation on diethylaminoethyl (DEAE)-Sephadex allowed for a comparison of the treated and control enzymatic activities using a common exogenous template. While no qualitative difference was evident, RNA polymerases I and II isolated from 3 methylcholanthrene-treated rats again were more active than control, indicating an effect of the polycyclic hydrocarbon at the level of the enzyme. PMID- 7471313 TI - A characterization of the activity of alpha-1,3,5-triglycidyl-s-triazinetrione, a novel antineoplastic compound. AB - To extend initial results on the antineoplastic activity of alpha-1,3,5 triglycidyl-s-triazinetrione (TGT, NSC 296934), a novel triepoxidic derivative, this compound was tested in a series of murine transplantable tumors. Repeated daily treatments with well-tolerated systemic doses of this chemical produced substantial retardation in tumor growth and significant prolongation of survival in the line 16 mammary, M5067 ovarian, and Madison 109 lung carcinomas and in mFS6 fibrosarcoma. Very marked activity was also seen in the P815 mastocytoma, B16 melanoma, line 38 colon carcinoma, and an intracerebrally transplanted ependymoblastoma, with high proportions of cures after one or two injections in IP transplanted SL2 lymphoma and line 26 colon carcinoma. It is concluded that the high level of antineoplastic effectiveness and the wide spectrum of TGT activity together with its novel structural characteristics could be of clinical significance. PMID- 7471314 TI - Clinical pharmacology of tamoxifen and N-desmethyltamoxifen in patients with advanced breast cancer. AB - Serum concentrations of tamoxifen and its metabolite, N-desmethyltamoxifen (DMT) were determined in six post-menopausal patients with advanced breast cancer. Following a single 10-mg dose PO parent drug was detected in the serum, with a peak concentration of 17.5 ng/ml. Concentrations of the N-desmethyl metabolite were below the limit of detection (less than 2.5 ng/ml). After 21 days' oral therapy with 10 mg b.i.d. the serum concentration of tamoxifen had increased ten fold, while DMT was now present in comparable amounts. Two patients were further studied for a longer time period. There was little change in the serum concentration of tamoxifen, while the DMT increased two fold above its value at 21 days. PMID- 7471316 TI - Early detection of doxorubicin cardiotoxicity by M-mode echocardiography. AB - The influence of increasing doses of doxorubicin on the heart was examined in 30 patients with solid tumors, M-mode echocardiography being used to evaluate left ventricular contractility. The function of the left ventricle remained normal in 26 subjects, whereas four patients had evidence of cardiotoxicity after cumulative doses of 220, 380, 420, and 450 mg/m2. Transient overt heart failure was noted in one subject only. Doxorubicin cardiotoxicity can be detected by M mode echocardiography, a simple and non-invasive technique, prior to the appearance of overt congestive heart failure. Patients demonstrating left ventricular dysfunction are probably not candidates for receiving further therapy with anthracycline antibiotics. Limitation of M-mode echocardiography include a 28% incidence of inadequate studies in this group of patients, and a relative inaccuracy of the technique in evaluating patients with prior myocardial infarction. PMID- 7471315 TI - Histopathological studies on antitumor effect of sporamycin. Cell-mediated immunity against allogeneic tumor-bearing mice. AB - Mice that had received transplants of sarcoma-180 followed by treatment with sporamycin were examined histopathologically at periodic intervals. A marked degeneration of tumor cells was observed at an early stage after the administration of sporamycin, but the degeneration subsequently ceased and regrowth of the tumor was seen. Marked infiltration of lymphoid cells, granulation tissue, and fibrosis was seen in the stroma or surrounding tissue of the tumor at a late stage after the administration of sporamycin, and the regression of tumor cells became marked. With a few exceptions the mice were completely cured by about the 40th day. In the peripheral lymphoid tissues, a transitory decrease in the number of cells was observed after the administration of sporamycin, but this was followed by regeneration of the cells, followed by a marked increase in the B cell system. On the other hand, lymphoid cell depletion of the thymus had persisted. Transplantation of intact sarcoma-180 to mice preliminarily inoculated with sporamycin-treated sarcoma-180 cells resulted in inhibition of tumor growth in most of the mice, and qualitatively the same tissue reactions as those in mice cured of sarcoma-180 by sporamycin were seen. The results suggest that enhancement both of antigenicity of the tumor (cells) and of the subsequent immune response of the host by sporamycin may be involved in the cure of the experimental tumor. PMID- 7471317 TI - Histological differentiation in human breast cancer is related to steroid receptors and stromal elastosis. AB - In 503 cases of human breast cancer, the stroma reaction of elastosis was investigated with respect to histological differentiation, pathological and biochemical prognostic factors, and steroid receptor (SR) content. Unlike perivascular elastosis, gland-related (ductal + interstitial) elastosis was not related to age, menopausal status, and number of pregnancies of each patient, and could thus be considered a histological feature characteristic of mammary cells. Elastosis was encountered most frequently in histologically differentiated lesions and in lesions of histoprognostic grades I and II (low degree of malignancy). Elastosis-positive lesions thus seem to constitute a good prognosis. Elastosis was related to the presence of estrogen and progestin receptors (ER and PR), and in menopausal patients it was observed mostly when both ER and PR were present concurrently, i.e., under conditions reflecting the hormone-dependence of neoplastic cells better than the presence of ER alone. Taken together, these results suggest that the presence of hormone-dependent cells in breast carcinomas can be demonstrated by both biochemical and morphological features. Since each of these factors has its own prognostic value, prognosis could probably be by the assessed more accurately if all these parameters were examined at the same time. PMID- 7471318 TI - Comparative toxicity of detorubicin and doxorubicin, free and DNA-bound, for hemopoietic stem cells. AB - We compared the toxicity of detorubicin (DET) and of doxorubicin (DOX) on the hematopoietic stem cells in C57BL6J mice by means of the CFUS and CFUC assays. On an equimolar basis DET appears to be less toxic than DOX for both the pluripotent stem cells and the granulocytic progenitor cells. Moreover, the administration of these anthracyclines as DNA complexes leads to a decreased toxicity to the pluripotent stem cells, while no such attenuated toxic effect is observed in committed stem cells. PMID- 7471319 TI - Amphotericin B plus combination chemotherapy for extensive non-small cell bronchogenic carcinoma. AB - Sixteen patients with extensive non-small carcinoma of the lung were treated with methotrexate, vincristine, cyclophosphamide, and adriamycin. Cyclophosphamide and adriamycin were administered after pretreatment with amphotericin B. Amphotericin B-related toxicity was mild; cytotoxic chemotherapy-related toxicity was tolerable. The partial response rate was 12.5% and median survival was between 19 and 20 weeks. Response rate and survival were not superior to those of a similar drug combination lacking amphotericin B. PMID- 7471320 TI - A comparison of aspects of the maternal speech environment of retarded and non retarded children. AB - This study was concerned to explore aspects of the maternal speech environment of mentally retarded children. Eight mothers were observed in interaction with their retarded children and compared with mothers on non-retarded infants who had been matched in terms of various socio-economic indices and language behaviour. The mothers of the retarded children were found to be significantly more stimulating vocally, more responsive and less directive than the controls. The findings are discussed in terms of possible reasons for such differences and the implications for the child. PMID- 7471321 TI - A behaviour management course for a group of mothers: the importance of the course setting for effective use of available resources. AB - Many patients experience difficulty in managing aspects of their young children's behaviour; and yet personnel with the psychological skills to resolve these problems are only available to a small proportion of parents, usually after a period of serious escalation of the difficulties. This article describes an attempt to make economical use of an educational psychologist's time by providing a short course for a volunteer group of mothers whose children attended a pre school group. The educational orientation of the centre is shown to facilitate the transmission of the required skills and knowledge to the mothers, suggesting both a desirable form of organization for such centres and a specific activity which could profitably take place within them. PMID- 7471322 TI - Parent support groups for the families of mentally handicapped children. AB - This paper describes a model of parent support group which has, as its aim, the provision of an effective clinical service to the handicapped child and his or her family. The aims and objectives of the group are outlined together with an evaluation of the effectiveness of the group after a period of 6 months. Conclusions are drawn as to the advantages of such a group and the importance of seeing the behaviour of the parent as an appropriate focus for intervention, is stressed. PMID- 7471323 TI - Community influences on breast feeding. AB - The incidence of breast feeding in Camden and Paddington is reported for two periods, 1974-75 and 1976-77. In 1976 a programme was begun to encourage breast feeding by increasing contact and support by the health visitor and child health visitor and child health clinic antenatally and postnatally. Fifty-eight per cent of mothers in Camden and 56% of mothers in Paddington breast fed at 3 weeks in 1976-77 compared with 24% (Camden) and 20% (Paddington) in 1974-75. The incidence of breast feeding in the experimental areas was compared with that in a similar control area in which no intervention occurred to investigate whether this increase reflected a widespread increase in the popularity of breast feeding or resulted from intervention. In 1976-77 more mothers in the experimental, than in the control area who breast fed on discharge continued to do so during the first 5 months. Of the mothers who were discharged breast feeding 88% in Camden and 73% in Paddington compared with 57% in the control area were still doing so at 6 weeks. The pattern of home visiting by the health visitors and attendance at the child health clinic was also investigated. More antenatal and postnatal home visits, more clinic visits and earlier home visits took place in the experimental areas compared with the control area. We conclude that the health visitor and child health clinic have an important role to play in the management of lactation. PMID- 7471324 TI - Patterns of intellectual change in the dementing school child. AB - Four children with intellectual deterioration are discussed. The importance of a developmental and educational history and sequential psychological testing is emphasized. Progressive visual impairment, clumsiness and increasingly poorly controlled fits should alert teachers and doctors to the possibility of associated intellectual deterioration. As well as global loss, tests of memory and mental arithmetic, manipulative and perceptual skills were the most sensitive index of dementia. We propose a further subcategory of the present codes of classification of childhood psychiatric disorders. PMID- 7471325 TI - Correlates of low birth weight. AB - In order to provide a base line for long term follow-up, the background, methodology and major conclusions of a prospective study of low birth weight infants (less than or to equal to 2500 g) and their matched controls (greater than 2500 g) were studied and the results presented. The low birth weight babies were disadvantaged in respect of a number of bio-social parameters, had an excess of maternal and pregnancy disease conditions and a more unfavourable outcome in other pregnancies. The cohort was used to try and evolve a better definition of impaired fetal growth. The relative merits of using weight, head circumference, length, ratio of anthropometric measurements, relative centile position of anthropometric measurements in the same baby, cluster analysis techniques and clinical morbidity as estimates of impaired fetal growth were analysed: none defined impaired fetal growth more accurately than weight for gestational age. Methods of estimating gestational age and physical characteristics relating to growth and gestational age, as well as a number of biochemical and neonatal measurements, are described. PMID- 7471326 TI - The treatment of intractable encopresis: a team intervention approach. AB - Childhood encopresis is discussed in terms of problems of definition, approaches to management in relation to symptoms and underlying mechanisms, and efficacy of treatment. Case histories of six intractable clinic cases are examined and an intensive team intervention programme described. Difficulties in carrying through a consistent plan are outlined: the key worker providing an essential motivational role towards participant staff, parents and child. PMID- 7471328 TI - Quantification of myocardial ischemia and infarction by left ventricular imaging. PMID- 7471327 TI - Two-dimensional echocardiography and infarct size: relationship of regional wall motion and thickening to the extent of myocardial infarction in the dog. PMID- 7471329 TI - Quantitative detection of regional left ventricular contraction abnormalities by two-dimensional echocardiography. I. Analysis of methods. AB - Different approaches to the quantification of regional left ventricular (LV) function from two-dimensional echocardiographic (2-D echo) images were assessed for their ability to optimize interobserver reproducibility in a heterogeneous patient population and to minimize the variability of regional function observed in a homogeneous normal population. Areas, hemiaxis and perimeter measurements were examined, as were the effect of the degree of image subdivision into halves, quadrants or octants. Each approach was also tested using both a fixed and a floating frame of reference for the definition of a regional-axis system. The area method was consistently superior to either linear method in optimizing both reproducibility and variability. Reproducibility decreased inversely with the degree of subdivision. The axis-system frame of reference had no effect on reproducibility. The floating-axis system yielded the same variability as the fixed system for short-axis sections at the mitral valve level, but slightly less variability for a papillary muscle level section. We conclude that area-based methods are superior for the evaluation of regional LV function with 2-D echo, but the degree of subdivision of the image and the frame of reference chosen do not greatly affect reproducibility or variability and should be chosen based on their performance in a well-defined clinical population. PMID- 7471330 TI - Quantitative detection of regional left ventricular contraction abnormalities by two-dimensional echocardiography. II. Accuracy in coronary artery disease. AB - The quantitative approaches to the assessment of regional left ventricular (LV) function described in the preceding paper were applied in a well-defined population of patients with coronary artery disease. Two groups were chosen by electrocardiographic and angiographic criteria: group 1 had infarction and regional wall motion abnormalities and group 2 had no infarction and normal wall motion. Sensitivity to detect wall motion defects, specificity to correctly categorize normal segments, and overall predictive accuracy were evaluated for each two-dimensional echocardiographic approach. In addition, the ability of each method to localize regional contraction defects properly was evaluated. Area methods yielded better predictive accuracy than linear methods (87-95% vs 76 84%). No significant differences in accuracy were noted between quadrant and octant approaches. The fixed external-axis system was superior to a floating one for localizing contraction defects. We conclude that an area-based method, using a fixed-axis system and either octant or quadrant image subdivision, provides the best combination of predictive accuracy in categorizing LV segments as normal or abnormal and the greatest ability to localize LV regional abnormalities. PMID- 7471331 TI - Repetitive responses to single ventricular extrastimuli in patients with serious ventricular arrhythmias: incidence and clinical significance. AB - Electrophysiologic studies were carried out in 85 patients with serious ventricular arrhythmias: 44 with recurrent sustained ventricular tachycardia (group A), 16 with recurrent nonsustained ventricular tachycardia (group B), and 25 with recent prehospital ventricular fibrillation not associated with acute myocardial infarction (group C). Programmed ventricular stimulation from the right ventricular apex included premature stimulation during normal sinus rhythm, atrial pacing, and ventricular pacing, as well as brief bursts of rapid ventricular pacing (RVP). A repetitive ventricular response (RVR) was defined as one or more non-stimulated premature ventricular depolarizations in response to a single paced premature ventricular depolarization during normal sinus rhythm or atrial pacing. RVRs were observed in seven of 44 (16%) group A patients, one of 16 (6%) group B patients, and three of 25 (12%) group C patients. In contrast, single and double premature ventricular stimuli during ventricular pacing and/or bursts of RVP resulted in the reproducible initiation of ventricular tachycardia in 40 of 44 (91%) group A patients, 10 of 16 (63%) group B patients, and 19 of 25 (76%) group C patients. We conclude that RVRs to single ventricular extrastimuli during normal sinus rhythm or atrial pacing are rare, and therefore are an insensitive index of susceptibility to serious ventricular arrhythmias in these patients. PMID- 7471332 TI - The clinical significance of the repetitive ventricular response. PMID- 7471333 TI - Initiation of sustained ventricular tachyarrhythmias in a canine model of chronic myocardial infarction: importance of the site of stimulation. AB - The importance of the site of stimulation to the initiation of sustained ventricular tachyarrhythmias was determined in 24 adult mongrel dogs. Studies were performed 3-30 days after two-stage occlusion of the mid- or distal left anterior descending coronary artery, modified by a reperfusion stage. Unipolar cathodal stimuli of twice-threshold intensity and 2 msec duration were introduced at five to 24 sites in each dog in the distribution of occluded and nonoccluded vessels. Strength-interval curves were constructed from 232 measurements at these sites and local properties of excitability and refractoriness were correlated with the ability to initiate arrhythmias. All dogs had sustained ventricular tachyarrhythmias inducible from at least one site. Intramyocardial sites with normal excitability and refractoriness within 2 cm of an area of infarction were most often successful (27 of 44, 61%) in the initiation of sustained arrhythmias. Less successful sites included normal left ventricular plunge electrode sites less than 2 cm from an area of infarction (eight of 32, 25%) (p = 0.002), left ventricular plunge electrode sites within an area of infarction (20 of 103, 19%) (p less than 0.001), normal right ventricular sites (five of 24, 21%) (p less than 0.001), and endocardial catheter sites (six of 29, 21%), (p less than 0.001). These findings suggest that local properties of excitability and refractoriness at the site of stimulation, as well as anatomic and geometric factors, may be critical in the initiation of sustained ventricular tachyarrhythmias using the technique of programmed electrical stimulation. PMID- 7471334 TI - Coronary surgery after recurrent myocardial infarction: progress of a trial comparing surgical with nonsurgical management for asymptomatic patients with advanced coronary disease. AB - A randomized trial of surgical vs nonsurgical management was carried out in men 60 years of age or younger who had recovered from a recurrent myocardial infarction. Of 205 patients considered, 100 had few or no symptoms and had coronary vessels favorable for bypass grafting; these patients fulfilled the trial conditions and were randomized (50 surgical and 50 nonsurgical). In 41 patients (elective nonsurgical group), randomization was not considered justifiable because of relatively unfavorable coronary anatomy or severe left ventricular dysfunction. Nineteen patients had elective surgery because of disabling angina despite full medical treatment or because of significant left main coronary stenosis. In 45 patients, coronary angiography was not undertaken because of medical contraindications or reluctance of the patient to enter the study. Actuarial survival curves (mean follow-up 4.5 years) show an annual mortality rate of 3-4% per year for all investigated patients, and no advantage for the randomized surgical over the randomized nonsurgical group. The results suggest that in the absence of disabling angina or left main coronary artery stenosis, coronary artery surgery need not be advised for survivors of recurrent infarctions who have severe coronary artery disease. Moreover, the prognosis for the group of patients not treated surgically appears to be better than has been previously described. PMID- 7471335 TI - Identification of left ventricular thrombi in man using indium-111-labeled autologous platelets. A preliminary report. PMID- 7471336 TI - Significance of coronary arterial thrombus in transmural acute myocardial infarction. A study of 54 necropsy patients. AB - In 54 necropsy patients with transmural acute myocardial infarction (AMI) and coronary arterial thrombi, histologic sections of coronary arteries that contained the thrombi were examined by video-planimetry to determine if the amount of luminal narrowing caused by thrombi was comparable to that produced by underlying atherosclerotic plaques, and to determine the amount of luminal narrowing by plaques immediately proximal and distal to the thrombi. The 54 coronary arteries in the 54 patients were narrowed 33-98% (mean 81%) by atherosclerotic plaque alone in cross-sectional area at the site of the thrombus (occlusive in 47 and nonocclusive in seven), from 26-98% (mean 75%) within the 2 cm segment proximal to the thrombus, and from 43-98% (mean 79%) within the 2-cm segment distal to the thrombus. Of the 54 arteries, 52 (96%) were narrowed 76-98% in cross-sectional area by atherosclerotic plaque alone at or immediately proximal or distal to the thrombus and 26 (48%) were narrowed 91-98% by plaque alone. The thrombi were 0.1--6.0 mm2 (mean 1.4 mm2) in cross-sectional area and the underlying atherosclerotic plaques were 3.0-21.0 mm2 (mean 8.7 mm2). Thus, among necropsy patients with transmural AMI, coronary thrombi occur at sites already severely narrowed by atherosclerotic plaques. PMID- 7471337 TI - Reduced coronary dilatory capacity and ultrastructural changes of the myocardium in patients with angina pectoris but normal coronary arteriograms. AB - Hemodynamic and metabolic studies were performed in 15 patients without heart disease (controls, group A), in 21 patients with typical stress-induced anginal pain but normal coronary and left ventricular angiograms (angina pectoris with normal arteriogram, group B), and in 10 patients with angiographically proved coronary artery disease (CAD, group C). Coronary dilatory capacity, determined by measuring total myocardial blood flow at rest and during maximal coronary vasodilatation (dipyridamole, 0.5 mg/kg i.v.), was markedly reduced in group B and C patients. In group B patients, left ventricular catheter biopsy specimens revealed no evidence of small-vessel disease, but did show histologic alterations of mitochondria. During atrial pacing, the control subjects showed no changes in myocardial lactate uptake, whereas in group B patients, myocardial lactate production occurred. In contrast to controls, patients in group B showed a significant decline in ejection fraction and circumferential fiber shortening during isometric exercise. These findings suggest that myocardial ischemia is the cause of angina pectoris in patients who have angina but normal coronary arteriograms. PMID- 7471338 TI - Imaging the inflammatory response to acute myocardial infarction in man using indium-111-labeled autologous platelets. AB - The feasibility of imaging the inflammatory response to acute transmural myocardial infarction in man using biologically active indium-111 (111In)-labeled autologous leukocytes was assessed in 36 patients. Indium-111 leukocytes (approximately 500 microCi) were injected i.v. 18-112 hours after the onset of chest pain. Cardiac imaging was performed 24 hours later with a mobile gamma camera. Twenty-one patients had positive images and 15 had negative images. The percent of positive images increased as the interval between infarction and 111In leukocyte injection shortened; all patients injected within 24 hours of infarction had positive images. Patients with positive images were injected with 111In leukocytes earlier after infarction (mean +/- SEM, 43 +/- 4 vs 63 +/- 7 hours; p less than 0.05) and were younger (53 +/- 2 vs 65 +/- 3 years; p less than 0.05) than those with negative images. Several other parameters that could possibly have affected the imaging results were examined and were not significantly different in patients with positive and negative images. These included peak serum creatine kinase, location of infarction, incidence of pericarditis, use of antiinflammatory drugs (aspirin and indomethacin) or membrane-active antiarrhythmic drugs (lidocaine and procainamide), peripheral leukocyte count and cell labeling efficiency. The function of the labeled cell was similar in patients with positive and negative images. Six patients with acute infarction serving as controls and given free 111In oxine and six patients with stable coronary artery disease given 111In leukocytes all had negative cardiac images. PMID- 7471339 TI - Frequency and clinical significance of failure to visualize the conus artery during coronary arteriography. AB - In patients with occlusion of the left anterior descending coronary artery (LAD) or right coronary artery (RCA), the conus artery, which arises at or near the origin of the RCA, often serves as a principal source of collateral circulation. Coronary arteriograms in 508 adult patients revealed that in 80.5% the conus artery was well visualized on the RCA angiogram, but that in 19.5% it was not adequately visualized due to injection of contrast distal to its origin. In the latter patients, the presence of conus-LAD or conus-RCA collaterals might therefore go undetected. Because the degree of distal filling via collateral circulation affects medical and surgical decisions, it is important to attempt to visualize the conus artery adequately whenever the LAD or RCA is obstructed. PMID- 7471340 TI - Beneficial effects of amrinone-hydralazine combination on resting hemodynamics and exercise capacity in patients with severe congestive heart failure. PMID- 7471341 TI - Prevention of nifedipine of abnormal coronary vasoconstriction in patients with coronary artery disease. AB - The hemodynamic and myocardial metabolic responses to the cold pressor test were studied in 15 patients with coronary artery disease and stable exertional angina. Every patient had abnormal coronary vasoconstriction during a control cold pressor test, even though 14 were receiving propranolol and 12 were receiving long acting nitrates. Mean coronary vascular resistance for the group increased 18 +/- 6% (SD) (from 0.80 +/- 0.12 to 0.94 +/- 0.20 mm Hg/ml/min, p less than 0.05); coronary sinus blood was unchanged, and the arterial-coronary sinus oxygen difference widened significantly (from 11.5 +/- 1.2 to 12.3 +/- 1.2 ml/100 ml, p less than 0.05). Four patients developed angina, accompanied in each instance by a negative arterial-coronary sinus lactate difference. After the administration of nifedipine (10 mg buccally) in 10 patients, the coronary vascular responses to a repeat cold pressor test were normal in each patient. Mean coronary sinus blood flow increased 27 +/- 12% (from 122 +/- 32 to 153 +/- 35 ml/min, p less than 0.05), coronary vascular resistance decreased 10 +/- 6% (from 0.85 +/- 0.16 to 0.76 +/- 0.16 mm Hg/ml/min, p less than 0.05), and the arterial-coronary sinus oxygen difference was unchanged. No patient experienced angina. The hemodynamic and coronary vascular responses to a repeat cold pressor test in five patients given placebo were unaltered from control responses. The protective effects of nifedipine were unaccompanied by any change in mean arterial pressure, left ventricular filling pressure or myocardial oxygen consumption either at rest or in response to the cold pressor test. Nifedipine appears to exert a selective antivasoconstrictor effect on the coronary vasculature. PMID- 7471342 TI - Influence of resting left ventricular function on the left ventricular response to exercise in patients with coronary artery disease. AB - First-pass radionuclide angiocardiography was used to access the left ventricular response to submaximal exercise in 150 patients with coronary artery disease (CAD). To test the hypothesis that resting systolic left ventricular function as determined by left ventricular ejection fraction (LVEF) was a predictor of the hemodynamic response to exercise, the study group included patients with a wide range of resting LVEF (0.12-0.82). The influences of resting LVEF, resting end diastolic volume, the number of diseased vessels, exercise end point and exercise double product on the change in LVEF (delta LVEF) during exercise were tested using multiple linear regression analyses. Resting LVEF was a highly significant predictor of the delta LVEF (p = 0.0001). Exercise duration was not related to either the resting LVEF or the delta LVEF. For the 112 patients in whom coronary anatomy was known, resting LVEF retained its significance as a predictor of delta LVEF (p = 0.002) even after adjustment for the significance of the extent of CAD (p = 0.0007) and the exercise end point (p = 0.06). Patients with normal resting LVEF showed the most profound decreases in LVEF, the highest frequency of new regional dysfunction and the largest relative increase in end-diastolic volume during exercise. As rest LVEF decreased, the magnitude of the delta LVEF and the frequency of new regional dysfunction decreased. Therefore, left ventricular function at rest is an important determinant of the direction and magnitude of change in left ventricular function during exercise. PMID- 7471343 TI - The response of left ventricular function and size to atrial pacing, volume loading and afterload stress in patients with coronary artery disease. PMID- 7471344 TI - The effects of large negative intrathoracic pressure on left ventricular function in patients with coronary artery disease. AB - Using first-pass radionuclide ventriculography, we evaluated the effects of decreases in intrathoracic pressure (Mueller maneuver [M] to -20 to -30 cm H2O) in 14 patients with five patients without coronary artery disease (CAD) and in 12 normal control subjects. In the patients with CAD, control ejection fraction was 0.53 +/- 0.06 (SEM) and control heart rate was 83 +/- 6 beats/min. These did not change during M. In the patients with CAD, control ejection fraction was 0.37 +/- 0.03 and heart rate was 82 +/- 7 beats/min. During M, heart rate did not change, but ejection fraction decreased to 0.33 +/- 0.33 (p less than 0.01). Examination of regional wall motion abnormalities showed akinesis of at least one myocardial segment in nine of 14 patients with CAD, but in none of the patients without CAD, nor in any of the 12 previously studied normal subjects (retrospectively analyzed). These data are consistent with the hypothesis that the Mueller maneuver acts to increase afterload placed on the left ventricle. Furthermore, in patients with CAD, the Mueller maneuver may have induced localized myocardial ischemia or unmasked areas of preexisting marginal function. PMID- 7471345 TI - Hemodynamics alterations induced by isoproterenol and pacing after aortic valve replacement with the Bjork-Shiley or St. Jude medical prosthesis. AB - Stress evaluation was carried out in 26 patients approximately 7 months after aortic valve replacement with Bjork-Shiley valves (13 patients) and St. Jude medical valves (13 patients). During isoproterenol infusion (0.3 micrograms/kg/min), cardiac output increased by a factor of 1.5 and aortic valve area decreased by 50% for both valve groups, while transvalvular gradients (rest: 7 +/- 2 vs 10 +/- 5 mm Hg, p greater than 0.05) increased by 42 +/- 18 vs 51 +/- 18 mm Hg (p greater than 0.05), i.e., to levels of moderate aortic stenosis. However, during pacing stress these values progressively decreased with rising heart rates. In other postoperative evaluations that included ergometric stress with isoproterenol and pacing, induced hemodynamic changes after aortic valve replacement were predictable and consistent with regard to both direction and magnitude, and they differed characteristically according to the type of stress used. We conclude that no functional differences between Bjork-Shiley and St. Jude medical valves can be claimed. Standardized evaluation with isoproterenol is a sensitive stress test of prosthetic valvular hemodynamics. Because of the apparent magnification of residual obstruction after aortic valve replacement, it has advantages over pacing. PMID- 7471346 TI - Anatomic correction of transposition of the great arteries: pre- and postoperative cardiac catheterization, with angiocardiography in five patients. AB - Six of eight patients survived anatomic correction of transposition of the great arteries and repair of associated cardiovascular anomalies at the Hospital for Sick Children, Toronto, Canada. Two of the six survivors also had tricuspid atresia, and continuity between the right atrium and the subaortic outlet chamber in these patients was provided by a valved external conduit, in addition to the arterial switch and coronary artery reimplantation. Five of the six patients have undergone complete postoperative clinical, hemodynamic and angiocardiographic investigation and form the basis of this communication. Our present indications for anatomic repair are discussed. PMID- 7471347 TI - Application of information theory to clinical diagnostic testing. The electrocardiographic stress test. AB - The inherent imperfection of clinical diagnostic tests introduces uncertainty into their interpretation. The magnitude of diagnostic uncertainty after any test may be quantified by information theory. THe information content of the electrocardiographic ST-segment response to exercise, relative to the diagnosis of angiographic coronary artery disease, was determined using literature-based pooled estimates of the true- and false-positive rates for various magnitudes of ST depression from less than 0.5 mm to greater than or equal to 2.5 mm. This analysis allows three conclusions of clinical relevance. First, the diagnostic information content of exercise-induced ST-segment depression, interpreted by the standard 1.0-mm criterion, averages only 15% of that of coronary angiography. Second, there is a 41% increase in information content when the specific magnitude of ST-segment depression is analyzed, as opposed to the single, categorical 1-mm criterion. Third, the information obtained from ECG stress testing is markedly influenced by the prevalence of disease in the population tested, being low in the asymptomatic and typical angina groups and substantially greater in groups with nonanginal chest pain and atypical angina. The quantitation of information has broad relevance to selection and use of diagnostic tests, because one can analyze objectively the value of different interpretation criteria, compare one test with another and evaluate the cost effectiveness of both a single test and potential testing combination. PMID- 7471348 TI - The genesis of gallop sounds: investigation by quantitative phono- and apexcardiography. PMID- 7471349 TI - Choice of electrocardiographic leads for recording the earliest QRS onset in noninvasive measurements. AB - A significant error may be introduced in intervals measured from the onset of the QRS if an electrocardiographic lead that does not record the earliest deflection is used. To ascertain to what extent the commonly used leads can be relied on to show the earliest QRS onset, 100 normal subjects and 219 patients with heart disease were studied by means of simultaneous recording of three leads: a right precordial lead chosen to show an rS configuration, lead II, and another limb lead chosen to show a qR configuration. Lead II most frequently showed a delayed QRS onset--in 34% of normal subjects and 36% of the patients. In the other limb lead the initial QRS deflection was delayed in 24% of the normal subjects and 23% of the patients. The QRS onset in the right precordial lead was never delayed in the normal subjects; however, it was delayed in this lead in 6% of the patients. The delays in each of the leads ranged from 5-20 msec. We conclude that while a right precordial lead is by far the most reliable single lead that can be used for interval measurements, simultaneous recording of a right precordial lead and a limb lead assures that recording of the earliest QRS onset in all cases. PMID- 7471350 TI - Correlation of the width of the QRS complex with the pathologic anatomy of the cardiac conduction system in patients with chronic complete atrioventricular block. AB - A study correlating the electrocardiographic findings and the histology of the atrioventricular (AV) conduction system was carried out in 14 cases with chronic complete AV block and in 13 cases without chronic complete AV block. Patients with chronic complete AV block were divided into two groups, based on the width of the QRS complex. The QRS complexes were narrow (less than 0.12 second) in four cases (group 1) and wide (greater than or equal to 0.12 second) in 10 cases (group 2). In group 1, the main lesion was located in the penetrating portion of His bundle (Hisp) in one heart, in the branching portion of the His bundle (Hisb) in another and in the combined regions of Hisb and the left bundle branch in two. Three of the four cases in group 1 had idiopathic fibrosis of the conduction system and one had calcific nodules in the central fibrous body. In group 2, the main lesion was located in Hisb in two cases, in the combined regions of Hisb and the right bundle branch in one, in the Hisb and in the bilateral bundle branches in two, and in the bilateral bundle branches in five. All cases in group 2 were of the idiopathic type, except case 5, which had calcific aortic stenosis. In 13 cases without chronic complete AV block the AV conduction system was histologically normal, except for slight-to-moderate aging changes in the His bundle or the bundle branches. Lesions of the Hisb, which is believed to be the "distal His" electrophysiologically, may induce complete AV block with narrow or wide QRS complexes, depending upon the severity of the lesions in Hisb or adjacent bundle branches. PMID- 7471351 TI - Surface recording of His-Purkinje activity on an every-beat basis without digital averaging. AB - Efforts to record evidence of electrical activity from the body surface originating in the His bundle or bundle branches have been reported since 1973. Almost exclusively, these techniques have required digital averaging of 50-100 sequential cardiac cycles. For immediate diagnostic, therapeutic and prognostic application, recording on an every-beat basis is highly desirable. This is especially important in instances of changing atrioventricular conduction, arrhythmias or less-than-constant RR intervals. Our object has been to develop a system for more nearly optimal noise reduction, to avoid the disadvantages of serial signal averaging, and to be able to record His-Purkinje activity in man on an every-beat basis. Using multiple parallel inputs wih linear amplification, additional logarithmic amplification, some bandpass filtering, and a logic circuit that ultimately examines and accepts or rejects a deflection as "true" signal, we can record, in most instances, on a beat-by-beat basis, this very valuable component of the cardiac electrical cycle. PMID- 7471353 TI - Gastric dilation during stimulation of cardiac sensory receptors. PMID- 7471354 TI - Sudden death and CAD. PMID- 7471352 TI - Electrophysiologic effects on combined autonomic blockade in patients with sinus node disease. AB - Thirty-two patients were studied before and after i.v. administration of 0.15 0.20 mg/kg of propranolol. Twenty-one of the 32 underwent combined autonomic blockade with the additional infusion of 0.04 mg of atropine. Twenty other patients with sinus node disease underwent electrophysiologic studies both before and after i.v. administration of 0.04 mg of atropine alone. Spontaneous cycle length, maximal corrected sinus node recovery time, sinoatrial conduction time, secondary pauses and intrinsic heart rate were measured. Secondary pauses were more common in those with abnormal intrinsic heart rates, and they did not correlate with changes in maximal corrected sinus node recovery time or sinoatrial conduction time. In patients with normal intrinsic heart rate, abnormal test measurements usually returned to normal after combined blockade (hypervagotonia); however, some patients showed a new abnormality after propranolol that was not reversible with atropine (catecholamine-dependent). Abnormal test responses in patients with abnormal intrinsic heart rate persisted or increased after combined blockade. We conclude that patients with sinus node disease may be categorized as (1) those with intrinsic sinus node disease; (2) those with normal intrinsic sinus node function but either relative hypervagotonia or catecholamine dependency; and (3) those with abnormal intrinsic sinus node function affected by vagal or catecholamine factors. PMID- 7471356 TI - Sulfinpyrazone alone? PMID- 7471355 TI - Transmyocardial platelet behavior in CAD. PMID- 7471357 TI - Effects of respiration and posture on PSVT. PMID- 7471358 TI - Exercise-induced alterations in left ventricular volumes and the pressure-volume relationship: a sensitive indicator of left ventricular dysfunction in patients with coronary artery disease. PMID- 7471359 TI - Diagnostic value of exercise electrocardiography and thallium myocardial scintigraphy in patients without previous myocardial infarction: a Bayesian approach. PMID- 7471360 TI - Comparative accuracy of electrocardiographic and vectorcardiographic criteria for inferior myocardial infarction. AB - Numerous criteria for the diagnosis of inferior wall myocardial infarction by electrocardiogram (ECG) and vectorcardiogram (VCG) have been published, but they have not been subjected to a systematic, independent evaluation. Accordingly, we studied 146 patients undergoing cardiac catheterization; 63 were normal and 83 had a history of infarction, a significant right coronary lesion and an inferior wall motion abnormality (inferior infarction group). No ECG or VCG criteria were considered in the designation of the two groups; rather, three sets of ECG and VCG criteria were evaluated for this purpose. Specificity was excellent (98-100%) and sensitivity was poor (4-34%) by all three sets of ECG criteria, but the 1949 ECG criteria of Meyers et al. are the least sensitive (4%, p less than 0.001). Specificity (90-100%) and sensitivity (82-84%) were very good by all three VCG criteria. The VCG criteria of Starr et al. gave no false-positive results in our normal group. Because of enhanced sensitivity, the overall accuracy of the VCG was higher than that of the ECG for the diagnosis of inferior infarction (90% vs 62%, p less than 0.001). We conclude that more recent ECG criteria for the diagnosis of inferior wall myocardial infarction are highly specific, but insensitive compared with VCG criteria. PMID- 7471361 TI - Usefulness of the Valsalva maneuver in management of the long QT syndrome. AB - Exercise or isoproterenol infusion may evoke ventricular arrhythmias in patients with the long QT syndrome. We examined the electrocardiographic effects of the Valsalva maneuver in eight patients with the long QT syndrome and nine healthy subjects. The Valsalva maneuver lengthened the QTc interval in both groups, but the lengthening was greater in the patients. In the patients who were having frequent attacks of ventricular tachycardia, the Valsalva-induced prolongation of the QTc interval was particularly remarkable and was associated with the development of T-wave alternans and short runs of ventricular tachycardia. Propranolol effectively suppressed the lengthening of the QTc interval during Valsalva strain and prevented Valsalva-induced ventricular arrhythmia. These results suggest that the Valsalva maneuver may be useful in evaluating the risk of ventricular tachyarrhythmias and the efficacy of drug treatment in patients with the long QT syndrome. PMID- 7471362 TI - Comparative effects of three calcium antagonists, diltiazem, verapamil and nifedipine, on the sinoatrial and atrioventricular nodes. Experimental and clinical studies. AB - Diltiazem, verapamil and nifedipine suppress sinoatrial (SA) nodal function in the excised rabbit heart. Clinically, however, their suppressive effect on the SA node is modified considerably by the reflex increase in sympathetic tone as a result of the fall in blood pressure caused by the vasodilating action of the calcium antagonists. Diltiazem, verapamil and nifedipine suppress atrioventricular (AV) nodal conduction and prolong refractory periods in the excised rabbit AV node. Clinically, diltiazem and verapamil exert a similar suppressive effect on the AV node and are useful for treating and preventing AV nodal reentrant tachycardia. Nifedipine, in clinically practical doses, has no antiarrhythmic properties, probably because of reflex activation of the sympathetic system secondary to its hypotensive effect, which is greater than that of the other two calcium antagonists. Diltiazem and verapamil may sometimes worsen AV conduction, especially in patients with conduction disturbances. Nifedipine, on the other hand, can be used as a coronary vasodilator with the least untoward effect on AV conduction. PMID- 7471363 TI - Natural history of chronic second-degree atrioventricular nodal block. AB - This report details our experience with documented chronic second-degree atrioventricular (AV) nodal block (proximal to His [H]) in 56 patients. Forty-six men (82%) and 10 women (18%), ages 18-87 years, were studied. Nineteen of the patients (34%) had no organic heart disease (including seven trained athletes) and 37 (66%) had organic heart disease. ECGs in all patients demonstrated episodes of type I second-degree block; five patients also had periods of 2:1 block. Prospective follow-up patients with no organic heart disease (157-2280 days, mean 1395 +/- 636 days) revealed one patient with clear indication for permanent pacing because of bradyarrhythmic symptoms (permanently placed on day 220 of follow-up). Two patients died nonsuddenly. In patients with organic heart disease (prospective follow-up of 60-2950 days, mean 1347 +/- 825 days), pacemakers were implanted in 10 patients, primarily for treatment of congestive heart failure in eight and syncope in two. Sixteen patients died -- three suddenly, seven with congestive heart failure, two of an acute myocardial infarction and four of causes unrelated to cardiac disease. In summary, chronic second-degree AV nodal block has a relatively benign course in patients without organic heart disease. In patients with organic heart disease, prognosis is poor and related to the severity of underlying heart disease. PMID- 7471364 TI - A simplified valve formula for the calculation of stenotic cardiac valve areas. AB - We have simplified the Gorlin formula and have compared our measurements of the aortic or mitral valve area, using the original Gorlin formula and the simplified valve formula in 100 consecutive patients. The valve area was measured by the simplified formula as cardiac output (l/min) divided by the square root of pressure differences across the valve. In patients with aortic stenosis of varying severity there were excellent correlation between the original Gorlin formula and the simplified formula (r = 0.96, y = 0.99x + 0.01, SEE = +/- 0.10, p less than 0.001). The correlation was unchanged when the peak gradient was used instead of the mean gradient in the simplified formula. Excellent correlation was also seen in patients with mitral stenosis of varying severity (r = 0.94, y = 0.97x - 0.02, SEE = +/- 0.19; p less than 0.001). The simplicity of the formula makes it easy to memorize and use. PMID- 7471365 TI - Accuracy of angiographic determination of left main coronary arterial narrowing. Angiographic--histologic correlative analysis in 28 patients. AB - To evaluate the accuracy of coronary angiography in identifying severe narrowing of the left main coronary artery (LMCA), the degree of narrowing observed by angiography was compared to that observed at necropsy in 28 patients with symptomatic coronary heart disease in whom angiography had been performed within 40 days of death. The angiograms were evaluated independently by three experienced angiographers. In 20 of the 28 patients (71%), the degree of narrowing of the LMCA was either underestimated (13 patients) or overestimated (10 patients) by two or three of three angiographers; of 84 angiographic judgments made by the three angiographers in the 28 patients, 54 (64%) were underestimates (33 judgments, 39%) or overestimates (21 judgments, 25%) of the degree of LMCA narrowing. Of 12 LMCAs narrowed 76-100% in cross-sectional area at necropsy, six were underestimated at preoperative angiography by two or three of three angiographers; of 12 LMCAs narrowed 51-75% in cross-sectional area at necropsy, all 12 were either under- or overestimated angiographically by two or three of three angiographers; of four LMCAs narrowed 26-50% in cross-section at necropsy, two were overestimated by two of three angiographers. Thus, angiographic determination of degrees of narrowing of the LMCA during life is subject to considerable error. The angiographic errors appear to have resulted primarily from the presence of atherosclerotic plaque in the LMCA and an insufficient number of angiographic projections. PMID- 7471366 TI - Automatic processing of cineventriculograms for analysis of regional myocardial function. PMID- 7471367 TI - Comparative accuracy of apical biplane cross-sectional echocardiography and gated equilibrium radionuclide angiography for estimating left ventricular size and performance. PMID- 7471368 TI - Esophageal echocardiographic left ventricular anterolateral wall motion in normal subjects and patients with coronary artery disease. AB - Esophageal echocardiography was developed for recording left ventricular anterolateral wall (LVAW) echocardiograms and was applied clinically to 14 normal subjects and 21 patients with coronary artery disease. LVAW echocardiograms were obtained satisfactorily in 11 of 14 normal subjects (75%) and 20 of 21 patients (95%) with coronary artery disease. LVAW echocardiograms were obtained by conventional anterior echocardiography in eight of 21 patients (38%) with coronary artery disease. In 11 normal subjects, mm/sec (mean 34.3 +/- 5.2 mm/sec); and diastolic wall thickness ranged from 9-12.5 mm (mean 11.2 +/- 0.7 mm). In 20 patients with coronary artery disease, LVAW motion obtained by esophageal echocardiography was classified into five groups according to the excursion, and the findings were in good agreement (80%) with those obtained by left ventriculography. Classification of LVAW motion by conventional echocardiography agreed with that of left ventriculography in only three of eight patients, although all eight patients had abnormal LVAW motion by the conventional method. In all patients except one, whose LVAW echocardiograms were obtained by conventional echocardiography, excursion was much less than that obtained by esophageal echocardiography. We conclude that the projection of an ultrasonic beam from the intraesophageal transducer is a better approach for accurate measurement of LVAW motion. PMID- 7471369 TI - Diagnosis of tricuspid regurgitation by contrast echocardiography. PMID- 7471370 TI - Contrast M-mode ultrasonography of the inferior vena cava. AB - Contrast M-mode and two-dimensional ultrasonography of the inferior vena cava were performed in 65 patients with various acquired and congenital cardiac disorders. After saline was injected into a peripheral arm vein, the inferior vena cava was visualized by both methods in 60 patients (92%). The M-mode approach was better for correlating the appearance of contrast within the inferior vena cava with the ECG. This precise correlation served as the basis for differentiating patients with tricuspid insufficiency (visualization of contrast after the QRS) from those with impaired right ventricular filling (visualization of contrast before the QRS) or arrhythmia. Results with the M-mode approach suggest that conditions other than tricuspid insufficiency may cause the appearance of contrast within the inferior vena cava. M-mode echocardiography should be used to further investigate patients with impaired right ventricular filling. PMID- 7471371 TI - Echocardiographic appearance of the Chiari network: differentiation from right heart pathology. AB - As echocardiography is being used more often, its value and accuracy are becoming more fully appreciated. Coincident with wider application of this imaging technique is the potential for identifying normal anatomic variants and their possible erroneous interpretation as pathologic states. In this report we describe the M-mode and two-dimensional echocardiographic features of a congenital remnant known as the Chiari network. This structure can present as a highly mobile, highly reflectant echo target that can be seen in several locations in the right atrium. We report here an index case that could be well examined echocardiographically and that was a cause of considerable concern due to the presence of congestive heart failure and a history of staphylococcal endocarditis. The presence of the Chiari network was confirmed pathologically. Subsequently, we found similar echocardiographic findings in 19 of 1248 patients (1.5%) studied in our laboratory. This congenital remnant, which is found pathologically in 2-3% of normal hearts, could be confused with valve disruption, vegetation or other mass lesion, particularly when associated with a suggestive clinical situation. PMID- 7471372 TI - Alterations in carotid arterial velocity-time profile produced by the Blalock Taussig shunt. PMID- 7471373 TI - Torsades de pointes: electrophysiologic studies in patients without transient pharmacologic or metabolic abnormalities. AB - Electrophysiologic studies were performed in 21 patients who had torsades de pointes. This ventricular tachyarrhythmia, characterized by rapid (200-250 beats/min) and irregular paroxysms and progressively varying QRS amplitude and polarity, occurred in the absence of electrolyte disturbance, antiarrhythmic drug therapy or acute ischemia. The QTc interval was prolonged in seven of 21 patients. Electrophysiologic study included ventricular pacing with the introduction of one to three extrastimuli and rapid ventricular pacing. The effect of i.v. procainamide or quinidine in these patients was also studied. Torsades de pointes was inducible n 19 of 21 patients. Induced episodes closely resembled spontaneous episodes. Torsades de pointes spontaneously progressed to ventricular tachycardia with a uniform morphology in three patients and to ventricular fibrillation in four. In eight patients, procainamide or quinidine converted torsades de pointes into typical reentrant ventricular tachycardia. Our data suggest that torsades de pointes in this setting may be a rapid reentrant ventricular tachycardia closely related to recurrent sustained ventricular tachycardia and a precursor to ventricular fibrillation and sudden death. PMID- 7471374 TI - Spontaneous termination of circus movement tachycardia using an atrioventricular accessory pathway: incidence, site of block and mechanisms. AB - The incidence, mechanisms and sites of block of spontaneous termination of circus movement tachycardia (CMT) using an atrioventricular accessory pathway (AP) were analyzed in 24 consecutive patients (17 with Wolff-Parkinson-White syndrome and seven with a concealed AP) who were not receiving antiarrhythmic drugs. Spontaneous termination of tachycardia occurred in 10 patients (105 episodes). A reduced "safety margin" of tachycardia was the only factor that was significantly more common in the patients who manifested spontaneous termination (p less than 0.01). The site of spontaneous block was located in the AP in six patients (50 episodes), atrioventricular node (AVN) in six patients (37 episodes) and His Purkinje system (HPS) in three patients (18 episodes). At least 14 mechanisms leading to block in the tachycardia circuit were identified. Labile conduction during tachycardia occurred at multiple sites (AVN, His bundle, bundle branches, and AP). Analysis of the duration of tachycardia before spontaneous termination showed a characteristic time pattern for block at each site, consistent with the autonomic and electrophysiologic changes that occur after induction of tachycardia. Spontaneous termination of CMT using an AP is a common phenomenon. Many mechanisms are involved, which are often complex and dependent on interplay of the electrophysiologic characteristics of the components of the tachycardia circuit. PMID- 7471375 TI - Atrial excitability and conduction during rapid atrial pacing. AB - Using temporary atrial wire electrodes placed at selected atrial sites, rapid atrial pacing at rates of up to 368 beats/min was used to study atrial excitability and conduction in seven patients who underwent open heart surgery. The threshold for atrial pacing was found to be an exponential function of pacing rate (r = 0.55, p less than 0.01), increasing threefold when the fastest pacing rates were compared with the slowest pacing rates (p less than 0.005). Atrial conduction times (measured from pacing to recording sites), prolonged during rapid atrial pacing both for studies conducted before institution of cardiopulmonary bypass (p less than 0.005) and for those done 7 days postoperatively (p less than 0.05). However, prolongation of conduction times always depended on achievement of a critically rapid pacing rate. During rapid atrial pacing, we observed a high incidence of alternans of the atrial electrogram (17 of 42 studies). Thus, human atrial excitability, conduction and electrogram morphology are not constant during pacing at rapid rates. Rather, at rapid pacing rates, there is depression of atrial excitability, prolongation of atrial conduction times and alternation in electrogram morphology. These findings have clinical relevance and theoretical implications for the understanding and treatment of rapid atrial rhythms. PMID- 7471376 TI - The nature of atriosinus conduction during rapid atrial pacing in the rabbit heart. PMID- 7471377 TI - Strength-interval relations in a chronic canine model of myocardial infarction. Implications for the interpretation of electrophysiologic studies. AB - Fifteen adult mongrel dogs underwent two-stage occlusion of the mid- or distal left anterior descending coronary artery and then a reperfusion stage. The dogs were studied 3-30 days later to determine strength-interval relations in a canine model of chronic myocardial infarction. These dogs were susceptible to the initiation of sustained ventricular tachyarrhythmias with the introduction of one, two or three ventricular extrastimuli. Using unipolar cathodal stimuli with a pulse width of 2 msec, strength-interval curves were constructed from measurements made at multiple sites in the distribution of occluded and nonoccluded vessels during drive pacing at a cycle length of 300 msec. At 56 normal sites, ventricular refractory periods measured at twice-diastolic excitability threshold approximated the relative refractory periods (mean absolute difference 3 msec), but were variably longer than effective refractory periods (mean difference 18 msec, range 4-29 msec). At 51 infarct sites, differences between ventricular refractory periods measured at twice-diastolic excitability threshold and both relative refractory periods (mean difference 13 msec, range -60 to +18 msec) and effective refractory periods (mean difference 28 msec, range 1-60 msec) were markedly disparate. These differences were further exaggerated after administration of i.v. procainamide. These findings suggest limitations in interpreting the results of programmed pacing studies performed using stimuli of twice-threshold intensity. PMID- 7471378 TI - Isointegral analysis of body surface maps: surface distribution and temporal variability in normal subjects. PMID- 7471379 TI - Body surface detection of delayed depolarizations in patients with recurrent ventricular tachycardia and left ventricular aneurysm. AB - In eight patients with chronic ventricular tachycardia and left ventricular aneurysms, we detected delayed ECG wave forms after the QRS complex from the body surface using a high-resolution ECG recorder, amplification and signal averaging. Delayed wave-form activity (D wave) extended a mean of 70 msec beyond the termination of th QRS complex. This delayed activity frequently extended to the limit of the recording window, and may thus continue throughout much of diastole. Antiarrhythmic agents never abolished the delayed activity; however, it was abolished by aneurysmectomy in four patients. Ventricular tachycardia did not recur after surgery in the four patients during a mean follow-up of 1 year. The D wave was not found in eight control patients who had chronic recurrent ventricular tachycardia nor in 11 of 12 who had aneurysms alone. The surface D wave can be readily and reproducibly detected by high-resolution electrocardiography and appears to be specific for patients with left ventricular aneurysms who also have chronic recurrent ventricular tachycardia. This delayed wave-form activity has been noted during catheter and surgical endocardial and epicardial mapping. It may represent persistence of the cardiac impulse in islands of myocardium and may be a manifestation of the delayed and fractionated activity, noted by previous investigators. PMID- 7471380 TI - Presidential address. AHA 53rd scientific sessions, Miami Beach, Florida, November 1980. Sure cures, quick fixes and easy answers. A cautionary tale about coronary disease. PMID- 7471381 TI - The effect of swimming on patients with ischemic heart disease. AB - Swimming is frequently recommended for cardiac rehabilitation, but little is known of its physiologic consequences in ischemic heart disease. Eight males who had had a myocardial infarction 8-17 months before the study were exercised to exhaustion or angina with 10 W/min-1 ramp on a cycle ergometer in sitting and supine positions. Oxygen uptake (VO2) was continuously measured to monitor the physiologic power requirement. All eight patients were taking beta blockers and four were taking digoxin. During sitting cycling, angina occurred in four and ST depression in five; during supine cycling, angina occurred in five and ST depression in six. VO2 was then measured while they swam at their own comfortable speed (mean 0.43 m/sec-1) in a swimming flume at water temperatures of 25.5 degrees C and 18 degrees C. In six, the water speed was gradually increased until they were limited by symptoms. Comfortable swimming at 25.5 degrees C was 87% (1.28 1/min-1) and at 18 degrees C 89% (1.30 1/min-1) of sitting peak VO2, while heart rates were 92% and 91% respectively. The mean peak VO2 and heart rate did not differ significantly between bicycle and swim tests (peak VO2 sitting 1.49 +/ 0.23, supine 1.42 +/- 0.24, 25.5 degrees C 1.60 +/- 0.17, 18 degrees C 1.52 +/- 0.19 1/min-1). Only two patients reported angina while swimming in warm water and one in cold water, although ST depression occurred in six in both swims. The subjective comfort and large muscle groups involved make swimming a good exercise, but the high relative energy cost and failure to identify ischemic symptoms indicate caution in cardiac patients, especially if their swimming skills are poor. PMID- 7471382 TI - Diagnostic quantification of CASS (coronary artery surgery study) clinical and exercise test results in determining presence and extent of coronary artery disease. A multivariate approach. AB - Multivariate linear discriminant function analysis on maximal exercise treadmill and angiographic data from 500 men with definite angina, 584 men with probable angina and 267 men with nonspecific chest pain identified independent predictors of presence and extent of coronary disease. We used the discriminant function to develop a clinical risk index and a clinical and exercise risk index for each patient subset. Probability curves were generated to predict the presence and extent of coronary disease. In definite angina cases, exercise testing provided more diagnostic information than clinical data alone. However, in the 10% of cases with the smallest risk indexes, half of the patients had coronary disease and one-quarter had multivessel disease. In men with probable angina, exercise testing added substantially more diagnostic information than clinical data. The probability of multivessel disease was reduced to less than 10% for 30% of patients with probable angina, an important diagnostic contribution. Exercise testing in men with nonspecific chest pain was of limited value because disease prevalence was already low. PMID- 7471383 TI - Improved method for determination of high-density-lipoprotein cholesterol I. Isolation of high-density lipoproteins by use of polyethylene glycol 6000. AB - We describe a modified method of precipitating low- and very-low-density lipoproteins with polyethylene glycol, for quantitation of high-density lipoprotein cholesterol. A 100 g/L concentration of polyethylene glycol 6000 in a buffered (pH 10) solution is used. Under these conditions precipitation of beta lipoproteins is complete. Values for HDL cholesterol as measured with this method fully correspond to those obtained on separating the lipoproteins by ultracentrifugation or with heparin-Mn2+, 46 mmol/L; are slightly higher than those obtained with heparin-Mn2+, 92 mmol/L; and are significantly higher than those determined after precipitation with polyethylene glycol, 100 g/L at pH 8.0. Comparison with results by the ultracentrifugation method showed excellent correlation (r = 0.957) with a good correspondence of results (slope 1.065 and y intercept -22.231); comparison with the heparin-Mn2+, 46 mmol/L, method gave r = 0.998 with slope of 0.985 and y-intercept 2.963. The method is unaffected by precipitation and centrifugation time and temperature (up to 24 h with 4-22 degrees C temperature for precipitation and between 10 and 30 min with 4-22 degrees C temperature for centrifugation). This extremely simple method is particularly suitable for routine analyses of many samples, even in small laboratories. PMID- 7471384 TI - Improved method for determination of high-density-lipoprotein cholesterol II. Enzymic determination of cholesterol in high-density lipoprotein fractions with a sensitive reagent. AB - A reagent is described for the colorimetric enzymic determination of high-density lipoprotein (HDL) cholesterol. The reagent can be used with HDL fractions isolated by the various methods of precipitation of low- and very-low-density lipoproteins we investigated. The considerable sensitivity obtained by use of Barham-Trinder's reaction allows the sample/reagent volume ratio to be decreased to 1:80, and major interferences thus eliminated. The response is linear from 100 to 2000 mg of HDL cholesterol per liter. The maximum CV obtained in precision tests was approximately 1% within series and approximately 3% between series. Most of the bilirubin interference is eliminated by adopting a reaction pH of 6.1. Because of its sensitivity, the reagent is particularly suitable for use with HDL fractions isolated after precipitation with polyethylene glycol 6000, which are characterized by a marked dilution. HDL cholesterol determination with the proposed reagent is accurate and precise. Values obtained are in line with those provided for by the Abell-Kendall method. The method can easily be automated. PMID- 7471385 TI - New approach to cystic fibrosis diagnosis by use of an improved sweat induction/collection system and osmometry. AB - A comprehensive equipment system is described that induces, collects, and analyzes sweat as a diagnostic procedure for cystic fibrosis. The safety and convenience of the iontophoretic induction phase have been improved by specially designed circuitry and electrodes. The problem of potentially serious error as a result of condensation within sweat-collection cups has been eliminated by use of a thermostatically controlled heated cup. The osmolality of the collected sweat is measured with a vapor-pressure osmometer. Data are presented to support the validity of osmolality as an alternative to electrolyte assay or electrical conductivity measurement and normal and abnormal ranges are provided. The complete system was tested in a pediatric hospital and data are presented to illustrate the relative advantages of the system with respect to safety, convenience, diagnostic reliability, and decreased potential for human error. PMID- 7471387 TI - Liquid-chromatographic analysis for urinary porphyrins. AB - Urinary porphyrins are separated, according to number of carboxyl groups, in a system consisting of a mu-Bondapak C18 stationary phase and a mobile phase of methanol and aqueous sodium phosphate (pH 3.5) in a linear gradient. The specimen is prepared simply by adjusting the pH of a 5-mL sample to 2.0 and removing solids by centrifugation. The eluted porphyrins are measured fluorometrically. Naturally occurring non-porphyrin fluorescent substances are eluted ahead of the porphyrins. Chromatography requires about 20 min, and a re-establishment of initial conditions requires an additional 15 min. PMID- 7471386 TI - Laser Doppler velocimetry vs heater power as indicators of skin perfusion during transcutaneous O2 monitoring. AB - Laser Doppler velocimetry (LDV) and heater power (HP) were compared as indicators of skin perfusion during transcutaneous oxygen tension (ptcO2) monitoring in adults. The LDV probe was fitted to a Radiometer ptcO2 attachment ring, which was fixed to the forearm. ptcO2, electrode temperature, HP, and LDV measurements were recorded continuously and simultaneously. LDV was more specific and sensitive to changes in cutaneous perfusion. Moreover, changes in perfusion were signalled four to 24 times faster by LDV than by HP. Changes in ptcO2 corresponded to changes in LDV more closely than to changes in HP in all experiments. HP was more specific and sensitive to changes in electrode and air temperatures. Once hyperemia was established at 44 degrees C and the electrode heater was turned off (HP = 0), LDV measurements frequently showed persistence of hyperemia (up to 25 min). Under these circumstances ptcO2 decreased linearly with decreasing electrode temperature in the range of 44-32 degrees C. LDV performance was referenced by monitoring velocity measurements taken on a cylinder revolving at known velocities. We conclude that LDV is a useful adjunct to ptcO2 monitoring and better than HP in assessing skin blood flow. It should enhance quality control of transcutaneous O2 monitoring. PMID- 7471388 TI - Rate-nephelometric inhibition immunoassay of phenytoin and phenobarbital. AB - A proposed rate-nephelometric inhibition immunoassay of phenytoin and phenobarbital in human serum involves the sequential addition to buffer of 42 microL aliquots of sample containing the hapten (drug) of interest, a hapten conjugate (drug-equine apoferritin), and specific antibody to the hapten. The drug and the drug conjugate compete for the binding sites on the antibody. The free hapten-antibody complex is soluble and so does not scatter light, whereas the complex of antibody and drug conjugate is insoluble and thus scatters light. The latter immunoprecipitation is competitively inhibited by free hapten. Thus the higher the concentration of free hapten present, the fewer immunoprecipitin complexes are formed, and the less light scatter. Precision, accuracy, linearity, analytical recovery, and comparison with patients' samples assayed with the DuPont aca were excellent. There was no significant interference from hemolysis, icterus, or lipemia. Many potentially interfering drugs and metabolites were checked for cross reactivity, with negative results. Reaction times range from 30 to 50 s. PMID- 7471389 TI - New method for determining lecithin and sphingomyelin in amniotic fluid. AB - We describe a new method of complete analysis for lecithin, lysolecithin, and sphingomyelin in amniotic fluid. The analysis is based on alkaline hydrolysis of the lecithins and the enzymic hydrolysis of sphingomyelin. The choline formed in each instance is enzymically phosphorylated with [gamma-32P]ATP to yield [gamma 32P]phosphorylcholine, which is isolated by anion-exchange chromatography and shown to be stoichiometrically related to lecithin, lysolecithin, and sphingomyelin. Other phospholipids do not interfere. Using this methodology, we developed three potential approaches to the assessment of fetal lung maturity: the ratio of lecithin plus lysolecithin (total lecithin) to sphingomyelin, the total lecithin concentration, and total lecithin as a percentage of total amniotic fluid phospholipids. All three indices were compared with the lecithin/sphingomyelin ratio obtained by a chromatographic procedure. Our data suggest that measuring the percentage of total lecithin may provide the best means of evaluating fetal lung maturity, but a final judgment must await clinical investigation. PMID- 7471390 TI - Gas-chromatographic assay of free phenytoin in ultrafiltrates of plasma: test of a new filtration apparatus and specimen stability. AB - I describe a gas-chromatographic procedure involving a nitrogen detector for "free" (or "unbound") phenytoin in the concentration range 0.2-4.0 mg/L in ultrafiltrates of plasma. The CV (n = 10) for the assay was 2.9% for a 0.49 mg/L concentration, 1.5% for 2.02 mg/L. The sensitivity was 0.05 mg/L. The ultrafiltration process ("Ultra-Free" filters) used to separate the free phenytoin fraction from the bound fraction was tested. The mean of and CV for free phenytoin concentrations measured in eight replicate serum filtrates were 0.68 mg/L and 15%; for eight replicate plasma filtrates the values were 0.42 mg/L and 2.9%. Thus serum is not suitable for this procedure. Storage of plasma before filtration under a wide variety of times and temperatures has an insignificant effect upon the filtration process. Some evidence was found of occasional protein "leakage" through the filters. PMID- 7471391 TI - Improved determination of D-glucaric acid in urine. AB - Using a simplex optimization procedure, we have elaborated a sensitive and reliable micromethod for determining D-glucaric acid, based on inhibition of glucuronidase activity by 1,4-D-glucarolactone, produced from D-glucaric acid by heating. The maximum attainable difference between glucuronidase activity in the reaction mixture with and without D-glucaric acid was used as an optimization criterion. From the optimization scheme, 1 mmol/L 4-nitrophenyl-beta-D glucuronide and pH 5.0 were found to be optimal conditions for determination of D glucaric acid with this test. In contrast to other similar methods, in which a logarithmic calibration curve is used, we calculate D-glucaric acid with a linear calibration curve by using the Dixon (Biochem. J. 55: 170, 1953) plot. The coefficient of variation for the method (within-run and between-day precision) was from 3.1 to 5.8%. Men excrete more D-glucaric acid than do women per 24 hours, but we saw no sex-related differences in excretion of D-glucaric acid when it was expressed in terms of urinary creatinine. PMID- 7471392 TI - Continuous-flow serum albumin determination by reaction with picrate ions, with use of a flow-through picrate ion electrode. AB - An automated potentiometric method for serum albumin determination by use of the picrate/albumin reaction is described. A continuous-flow system and a specially designed flow-through picrate ion electrode were used in making the measurements. Various factors affecting the reaction, such as pH, picrate ion concentration, and reaction time, were studied. Peak height in millivolts and albumin concentration were linearly related in the range 10-70 g/L. Both within-run and day-to-day, the CV for the method was about 2%. Analytical recovery of albumin added to serum samples ranged from 97.0 to 110.3%, averaging 102.2%. Results compare favorably with those by the established bromcresol green method. The proposed method is suitable for routine use and for screening tests. PMID- 7471394 TI - Rapid liquid-chromatographic determination of metformin in plasma and urine. AB - We describe an analysis for metformin in plasma and urine by use of cation exchange "high-performance" liquid chromatography. The assay requires only 0.5 mL of sample and involves pretreatment with trichloroacetic acid containing an internal standard (1-propylbiguanide), followed by centrifugation and injection into the chromatograph. The column eluent is monitored at 230 nm. Metformin and 1 propylbiguanide give retention times of 8 and 10 min, respectively. Within-day and between-day precision (CV) for the assay of plasma containing 2 mg (1.2 X 10( 5) mol) of metformin hydrochloride per liter was 3.6 and 3.9%, respectively. Similar precision was obtained for urine. The limit of detection is about 0.1 mg (6.0 X 10(-7) mol) of metformin hydrochloride per liter. Several commonly used drugs, including other oral hypoglycemic agents, do not interfere with this method. PMID- 7471393 TI - Enzymic determination of branched-chain amino acids. AB - Transamination of branched-chain amino acids to (alpha-oxoglutarate, catalyzed by leucine aminotransferase coupled to the glutamate dehydrogenase reaction, is used in an enzymic assay for determination of branched-chain amino acids in serum and tissue homogenates. The coefficients of variation of the method within-day and day-to-day are 2.4 and 6.5%, respectively. Analytical recovery of physiological concentrations of branched-chain amino acids added to serum is near 100%. Measurements in serum of healthy subjects revealed normal values similar to those found by use of other methods. During prolonged fasting the concentration of these amino acids in serum first increases, reaching a maximum by three days, followed by a successive decline. PMID- 7471395 TI - Liquid-chromatographic determination of 4-aminopyridine in serum, saliva, and urine. AB - We have developed "high-performance" liquid-chromatographic methods for determining 4-aminopyridine, an acetylcholine-releasing drug, in serum, saliva, and urine. As little as 1 microgram/L can be detected by extracting the alkalinized sample plus the internal standard (3,4-diaminopyridine) into dichloromethane, mixing the organic phase with 1-pentanol, evaporating the dichloromethane, and injecting the residue onto a reversed-phase column, where it is eluted with acetonitrile/methanol/aqueous ammonium carbonate, with detection at 245 nm. Analytical recoveries from serum averaged 86.7%. The CV at 50 micrograms/L was 2.9% (n = 8). For urine samples containing very high concentrations of 4-aminopyridine, we mixed urine and potassium carbonate in an automatic injector vial, extracted the drug into dichloromethane, centrifuged, and injected an aliquot of the extract into the chromatograph. Analytical recoveries averaged 92%, and the CV was about 2% for drug concentrations of 0.1-8 mg/L of urine. PMID- 7471396 TI - Fluorometric liquid-chromatographic determination of serum cortisol. AB - We describe a sensitive and specific liquid-chromatographic assay for serum cortisol, exploiting acid-induced fluorescence. Each analysis requires only 50 microL of serum, and chromatography is complete in 7 min. Analytical recovery of cortisol added to serum ranged from 90 to 105% and between-run precision (CV) from 6 to 8%. The lower limit of detection for cortisol is 10 micrograms/L, and linearity extends to 1600 micrograms/L. Numerous steroids and drugs tested did not interfere. PMID- 7471397 TI - Kinetic potentiometric determination of creatinine in serum with a picrate ion selective membrane electrode. AB - We describe a new kinetic method for potentiometric determination of creatinine in serum, based on the creatinine--picrate reaction in alkaline medium (Jaffe reaction). The reaction is monitored with a picrate-selective electrode, and the increase in electrode potential during 270 s is measured and related directly to the creatinine concentration. Small cation-exchange columns are used to separate creatinine from interfering substances. Analytical recovery of creatinine added to serum was 100.7%. Results for a series of samples compared well with results obtained with a spectrophotometric method (r = 0.994). PMID- 7471398 TI - The Worthington "ultrafree" device evaluated for determination of ultrafiltrable calcium in serum. AB - We evaluated a commercially-available disposable device ("Ultrafree," Worthington Diagnostics) for the anaerobic preparation of protein-free ultrafiltrates from serum for measurement of ultrafiltrable calcium. Sufficient filtrate for the analysis is obtained with 10 min from 0.2 to 0.4 mL of serum at room temperature. We assessed these ultrafilters with regard to permeability of calcium citrate, exclusion of proteins, frequency of leakage, and effect of temperature on results. Within-run and day-to-day coefficients of variation for human serum pools were 1.2 and 1.5%, respectively. Reference intervals (in mmol/L) for total (2.16-2.58), ultrafiltrable (1.44-1.67), dialyzable (1.25-1.41), and ionized (1.04-1.25) calcium have been determined for a healthy population of 69 women and 81 men, ages to 18 to 65 years. The device appears to be the most practicable yet available for use in making this measurement. PMID- 7471399 TI - Agar gel electrophoretic determination of glycosylated hemoglobin: effect of variant hemoglobins, hyperlipidemia, and temperature. AB - We investigated the effect of temperature, variant hemoglobins, and hyperlipidemia on determination of glycosylated hemoglobin by an electrophoretic method (Clin. Chem. 26: 1598-1602, 1980). We found that: (a) temperature variations ranging from 4 to 30 degrees C were without effect on results obtained by electrophoresis; (b) concurrent determination of glycosylated hemoglobin by electrophoresis and column-chromatography in blood specimens from 150 diabetic patients yielded almost identical mean values for both procedures when operations were carried out at 22 degrees C; (c) electrophoretic determination of glycosylated hemoglobin in whole-blood hemolysate was not affected by concentration of triglycerides; and (d) unlike column-chromatographic procedures, which underestimate the percentage of glycosylated hemoglobin in patients with hemoglobin S and C, the electrophoretic method accurately determined the proportion of glycosylated hemoglobin in these hemoglobinopathies. Evidently, electrophoresis on agar gel is an excellent alternative to cation-exchange column chromatographic methods for glycosylated hemoglobin. PMID- 7471400 TI - Measurement of hemoglobin A1 by liquid chromatography and by agar gel electrophoresis compared. AB - We compare measurement of total fast hemoglobin (HbA1) by "high-performance" liquid chromatography and by electrophoresis on agar gel. Blood samples were obtained from a diverse population (n = 222): offspring of two diabetic parents, diabetic patients with and without retinopathy, diabetic and non-diabetic pregnant women, patients in the coronary-care unit, and normal persons. Precision studies with a normal and an above-normal A1 sample resulted in overall CVs of 9.0% and 4.6% for the electrophoretic method and 4.4% and 2% for the chromatographic method. Linear regression analysis of values for total fast hemoglobin for the complete sample population and for each subgroup showed results of the electrophoretic method to be in excellent agreement with those by the chromatographic method. We conclude that the agar gel electrophoretic method offers a reproducible means for HbA1 determination that is comparable to the HPLC method in terms of accuracy and is highly suited for routine laboratory use. PMID- 7471401 TI - Clinical laboratory determination of phosphatidylglycerol: one- and two dimensional chromatography compared. AB - Reportedly, determination of several phospholipids in amniotic fluid, including phosphatidylglycerol and phosphatidylinositol, in addition to lecithin and sphingomyelin improves prediction of neonatal pulmonary function. In this study, we evaluated a two-dimensional technique for separating and measuring these phospholipids and compared it with a simpler one-dimensional procedure. The two dimensional technique was adapted to readily available commercial plates, and a preheating step was introduced to avoid shattering of the plates during charring. The Rf values, reproducibility of each technique, and the correlation between them were examined. Even though the one-dimensional technique is faster and less expensive, we recommend the two-dimensional method for clinical use because of better precision (CV for phosphatidylglycerol 15% vs 21%) and clearer results when relatively little phosphatidylglycerol is present. The one-dimensional procedure is unreliable when blood or meconium are present. In addition, interfering compounds co-migrate with phosphatidylglycerol, phosphatidylethanolamine, and phosphatidylserine in the one-dimensional technique. Before any one-dimensional lipid separation is adopted for clinical use, it should be critically compared to the two-dimensional procedure. PMID- 7471402 TI - Low results for inorganic phosphorus with the SMAC continuous-flow analyzer. AB - Serum inorganic phosphorus concentrations as measured with the SMAC are lower than those found with other methods. To resolve this problem we analyzed patients' specimens and performed analytical recovery studies with four different systems (SMAC, AutoAnalyzer II, aca, and the Fiske--SubbaRow method). With the SMAC, results for patients' specimens are significantly lower (p less than 0.0001) than with any of the other three methods. The SMAC recovered only about 87% of the added inorganic phosphorus. The value assigned to SMAC Reference I for inorganic phosphorus was 0.876 of the value obtained when the material was analyzed by the reference method. Thus there is a significant systematic error in the SMAC method for inorganic phosphorus determination, attributable to an erroneous inorganic phosphorus concentration assigned the SMAC calibration material by the supplier (Technicon). PMID- 7471403 TI - A multi-rule Shewhart chart for quality control in clinical chemistry. PMID- 7471404 TI - Age-dependence of immunoreactive trypsin concentrations in serum. PMID- 7471405 TI - Liquid-chromatographic analysis for urinary 4-hydroxy-3-methoxymandelic acid and 4-hydroxy-3-methoxyphenylacetic acid, and its use in investigation of neural crest tumors. PMID- 7471406 TI - Clinical evaluation of two thyrotropin radioimmunoassay kits: human serum matrix calibrators and bovine serum matrix calibrators. PMID- 7471407 TI - Evaluation of a new ferritin radioimmunoassay kit. PMID- 7471408 TI - Diluent pH and the stability of the thiol group in monothioglycerol, N-acetyl-L cysteine, and 2-mercaptoethanol. PMID- 7471409 TI - Nafcillin may cause a subtle pseudoproteinuria. PMID- 7471410 TI - Improved reliability of the liquid chromatography--electrochemical detection assay technique for measuring plasma epinephrine. PMID- 7471411 TI - Error in reporting unsaturated iron-binding capacity of serum. PMID- 7471412 TI - New PDP-8A computer program (DATAIO) for handling data from automated systems. PMID- 7471413 TI - Urinary steroid metabolite analysis: potential for renaissance. PMID- 7471414 TI - Use of glass beads to defibrinate serum in "microtainers". PMID- 7471415 TI - Simple automated system for continuous-flow determination of glycerol. PMID- 7471416 TI - Liquid chromatography of anticonvulsants in an inappropriately drawn (lipemic) serum sample. PMID- 7471417 TI - Screening for the Tay-Sachs carrier: a compromise program. AB - A modified program designed to screen for the Tay-Sachs carrier is presented in which testing is limited to one or both partners, as needed, in an Ashkenazi Jewish (Jews of central and eastern Europe ancestry) mating when there is a definite commitment toward having a child, or in the early stages of the pregnancy. Testing of unmarried individuals is discouraged. The approach maximizes individualization of both the medical and laboratory aspects of the program and promotes a positive and beneficial relationship between physician and clinical chemist. There is little involvement of the lay population or clergy, and no special sources of funding are required. Whereas most mass screening programs for the Tay-Sachs carrier have attempted to educate large numbers of the lay public to bring pressure upon the medical community, we have reversed this approach and taken the much easier course of educating small numbers of physicians to better care for their patients. The program has been used successfully in a moderate-size city in which communication lines between laboratory and physician are easily established. PMID- 7471418 TI - Esterolytic method for determination of heparin in plasma. AB - This assay for heparin is based on the heparin-accelerated rate of alpha-thrombin III. The rate or product formation from the residual active thrombin is inversely proportional to plasma heparin content. The assay can be performed manually, but our results were obtained with a discrete analyzer, the ABA-100. The assay is insensitive to concentrations of antithrombin III in plasma. Precision studies gave CVs of less than 10%. This assay was compared to the Dade Protopath heparin assay and a correlation coefficient of 0.90 was obtained (n = 62). The correlation between activated partial thromboplastin times and heparin concentrations (r = 0.67) was calculated frm results on 78 plasma specimens from 10 patients. PMID- 7471419 TI - Quantitative determination of apolipoproteins C-I and C-II in human plasma by separate electroimmunoassays. AB - Separate electroimmunoassays are described for measuring human plasma apolipoproteins C-I and C-II. Purified apolipoproteins C-I and CII were used in preparing monospecific antisera and as the primary standards. These assays are sensitive (maximal sensitivity, 20 ng), specific, rapid, precise (the within- and between-assay coefficients of variation for both assays were 5 and 8%, respectively), and accurate (accuracy was based on comparison of calculated and measured C-I, C-II, and C-III contents of an ApoC-containing column-eluent fraction) and are applicable to measurement of C-I and C-II polypeptides in whole plasma and density classes. However, plasma samples with triglyceride (triacylglycerol) concentrations greater than 6000 mg/L must be delipidized before analysis for C-II, as must those with greater than 12 000 mg/L before analysis for C-I polypeptide. Mean concentrations (and SD) of C-I in plasma of normolipidemic subjects and hyperlipoproteinemic phenotypes IIa, IIb, IV, and V were 60 (15), 70 (20), 100 (20), 100 (20), and 260 (94) mg/L, respectively. The corresponding C-II values were 40 (20), 43 (20), 68 (20), 65 (20), and 210 (70), respectively. C-I and C-II concentrations in patients with phenotypes IIb v, IV, or V significantly (p less than 0.001) exceeded those in normal persons or phenotype IIa. The observed correlations (r = 0.92 and r = 0.94) between triglyceride and C-I and C-II values suggest that these two polypeptides, like C III, are excellent plasma markers for assessing the state of triglyceride metabolism. PMID- 7471420 TI - Evaluation of pteroylglutamic acid and n-5-methyltetrahydrofolic acid reference standards in a new "No-Boil" radioassay for serum folate. AB - Serum folate, pteroylglutamic acid standards, and N-5-methyltetrahydrofolate standards extract differently during sample preparation for radiometric assay. B/B0 values for each material were determined before and after an alkaline extraction procedure. Serum folate most closely resembled N-5 methyltetrahydrofolate standards in that it exhibited a slightly greater ability to displace iodinated folate derivative in competition for beta-lactoglobulin binder. Pteroylglutamic acid standards displaced folate derivative less well, so that serum folate values were lower than when N-5-methyltetrahydrofolate was used as the standard material. PMID- 7471421 TI - Kinetic measurement of guanine deaminase in serum with a centrifugal analyzer. AB - We describe an enzymic, one-step kinetic method for determination of guanine deaminase (guanase, EC 3.5.4.3) in serum with a centrifugal analyzer. A combined enzyme-substrate system consists of the enzymes xanthine oxidase, catalase, and aldehyde dehydrogenase, the coenzyme NAD+, the substrate guanine, and ethanol in tris(hydroxymethyl)methylamine buffer, with KCl added as activator for aldehyde dehydrogenase. The method requires only 40 microL of sample. Guanase activity in 28 samples can be determined within 10 min by setting a 4-min lag period. The increase in absorbance at 340 nm is linearly proportional to the activity of guanase to 60 U/L. Within-run precision (CV) was 1.32 to 4.50% over the range studied. Day-to-day precision corresponds to CVs of 4.8 to 7.2% over the same range of guanase activity. The reference interval, as calculated from data on 25 healthy humans, was 0 to 1.02 U/L. The enzymic automated method shows good correlation with Caraway's (Clin. Chem. 12: 187, 1966) method (r = 0.949). PMID- 7471422 TI - Determination of Cu, Mn, Fe, and Ca in six regions of normal human brain, by atomic absorption spectroscopy. AB - A method is reported for measuring copper, manganese, iron, and calcium in brain tissue. We solubilized 100-mg samples of brain tissue with Soluene-350 and diluted the mixtures with methyl isobutyl ketone. Copper and manganese concentrations were measured by using a carbon rod atomizer, iron and calcium concentrations by using an air/acetylene and nitrous oxide/acetylene flame, respectively. The basal ganglia (caudate, putamen, globus pallidus, and substantia nigra) contained high concentrations of iron (10 to 19 mumol/g dry wt) as compared to concentrations found in the cortex (4 mumol/g) or the hippocampus (2.8 mumol/g). Copper exhibited a similar distribution pattern among the regions studied; 526 to 1140 mumol/g (dry wt) in the basal ganglia vs 496 mumol/g in the cortex and 426 mumol/g in the hippocampus. Manganese and calcium were uniformly distributed in the regions examined. PMID- 7471423 TI - Evaluation of a routine method for determination of calcium oxalate crystal growth inhibition in diluted urine samples. AB - We have evaluated a routine method for determination of the inhibition of calcium oxalate crystal growth in diluted urine samples. A crystallization inhibition index is calculated from the decrease in [14C]oxalate concentration in a metastable solution of calcium chloride and sodium oxalate after addition of seed crystals of calcium oxalate. The coefficient of variation between different determinations on the same sample is less than 1.5%, and there is good correspondence between indices obtained by this method and by a more laborious older method. We also studied the influence of different urinary constituents on the crystal growth rate in the system; whereas there was significant inhibition with citrate, chondritin sulfate, and pyrophosphate, no effects were observed with ordinary urinary concentrations of calcium, magnesium, and oxalate. The mean inhibition index was lower in a group of male patients with calcium oxalate stone disease than in a control group of normal men. PMID- 7471424 TI - Short- and long-term biological variation in analytes in urine of apparently healthy individuals. AB - Intra-individual, interindividual, and analytical components of variation for each of 10 urine analytes were estimated from results obtained on analysis of urine specimens from 10 apparently healthy young men. In spot urines, urea and osmolality showed strong individuality, but intra-individual variances were larger than interindividual variances for sodium, urate, phosphate, and glucose. Urea and osmolality displayed strong individuality in daily urines, whereas sodium (when results were expressed in output terms), urate, potassium, and total protein (when results were expressed in concentration units) had intra-individual variances larger than interindividual variances. In monthly urines, calcium and glucose, in concentration terms, and phosphate, in output terms, exhibited strongest individuality. Analytical variance was a significant percentage of the total variance for total protein and glucose; the analytical methods we used for these analytes were probably unsatisfactory. A series of analytical goals for imprecision, based on biological variation, were derived. Although these goals may not be applicable in all clinical situations, they represent the beginning of a data base in the literature. PMID- 7471425 TI - Parathyrin measured concurrently with free or total calcium in the differential diagnosis of hypercalcemia. AB - We assessed the diagnostic utility of measuring C-terminal immunoreactive parathyrin (i-PTH) concurrently with free (CaF) or total (CaT) calcium in four groups of patients: (1) 91 patients with histologically proven primary hyperparathyroidism; (2) seven patients without histological evidence of hyperparathyroidism; (3) 70 patients with non-parathyroid malignancies with hypercalcemia; and (4) 28 similar patients without hypercalcemia. In patients in Group 1, the use of either calcium measurement along with i-PTH increased diagnostic accuracy from 95 to 99%. In Group 2, values consistent with primary hyperparathyroidism were obtained for 21% of samples with i-PTH assay alone, 29% with i-PTH and CaT, and 12% with i-PTH and CaF. Surprisingly, 36% of samples from both malignancy groups (3 and 4) had above-normal i-PTH values. In patients with malignancy and hypercalcemia (Group 3) the following percentages were classified as possibly being hyperparathyroid: 26% with i-PTH assay alone; 31% with i-PTH and CaT, and 24% with i-PTH and CaF. Our data indicate that (a) about a fourth of patients with nonparathyroid malignancies also have evidence of hyperparathyroidism as judged by this (and probably all other) i-PTH methods based on C-terminal antibody specificity and that (b) measuring CaF and i-PTH rather than CaT and i-PTH gives a small increase in diagnostic accuracy. PMID- 7471427 TI - Test of radioimmunoassay validity in the presence of an isotope effect. AB - An isotope effect, manifested as delayed elution of the labeled moiety, was detected in CO-chromatography of testosterone with [1,2,6,7-3H]testosterone on Celite. The isotope effect was smaller for [1,2-3H]testosterone and even smaller, but still significant, for [7-3H]testosterone. Because of the isotope effect, the test of validity of a testosterone radioimmunoassay had to be modified. Instead of testing the constancy of values for specific activity in individual chromatographic fractions, the slopes of specific activities exhibited by mixtures of plasma testosterone (pooled specimens of plasma from men and from women) with [1,2-3H]testosterone were compared with that yielded by authentic testosterone mixed with the same tracer. Because the slopes were statistically indistinguishable, the assay of testosterone in these materials appears to be valid. We suggest that comparison of the slopes of specific activities be used as the basis of validity tests whenever such an isotope effect is present. PMID- 7471426 TI - Metabolic abnormalities associated with diabetes mellitus, as investigated by gas chromatography and pattern-recognition analysis of profiles of volatile metabolites. AB - Patterns of volatile metabolites in urine, as obtained by glass-capillary gas chromatography, were investigated by use of a nonparametric pattern-recognition method, in an effort to detect abnormalities associated with diabetes. We used threshold logic unit analysis on a data set consisting of normal subjects and those with diabetes mellitus, and could predict patterns for volatile metabolites as belonging to the proper class in 94.83% of the cases examined. In addition, a feature-extraction algorithm isolated those volatile constituents that are most useful in making the normal/diabetic classification. We used gas chromatography/mass spectrometry to identify important profile constituents. Finally, these same pattern-recognition methods indicated strong sex-related patterns in these volatiles. PMID- 7471428 TI - Positive interference by danazol in a testosterone radioimmunoassay kit procedure. AB - We describe a problem in measuring serum testosterone concentrations in the serum of women who are taking danazol. The Diagnostic Products Corporation testosterone radioimmunoassay gives very high apparent testosterone concentrations in these women and the value obtained depends on the volume of serum extracted per assay tube. Chromatography of the serum extracts on Sephadex LH-20, but not on Celite, before radioimmunoassay reduced these high apparent testosterone concentrations to less than that found in normal women. PMID- 7471429 TI - Prostatic acid phosphatase as measured with two radioimmunoassay kits in the detection of prostatic adenocarcinoma. AB - Serum prostatic acid phosphatase concentration was measured with two commercially available radioimmunoassay kits. Results were compared with histological evidence of prostatic adenocarcinoma obtained at autopsy in 33 patients. The serum assay did not differentiate significantly (p greater than 0.1) between patients with adenocarcinoma and those without. We conclude that the test, at least as performed by use of these kits, is of little value in the detection of occult disease. PMID- 7471430 TI - Preparation of whole blood for liquid scintillation counting. AB - Liquid scintillation counting of 3H-labeled whole-blood samples is severely impaired owing to quenching by blood pigments. In this study, dry oxidation and chemical solubilization followed by decolorization were the two general methods used to eliminate color quenching. Three blood volumes were examined: 0.25, 0.50, and 1.0 mL. Dry oxidation yielded complete recoveries of 3H label with counting efficiencies greater than 30% for up to 1.0 mL of blood. Although blood volumes larger than 0.25 mL can be used with chemical solubilization and decolorization, treatment of 0.25 mL of blood gave the highest counting efficiencies, with count rates comparable to those for 1.0-mL samples. PMID- 7471432 TI - Preincubation of serum aspartate aminotransferase with pyridoxal 5'-phosphate in the SMAC: comparison with revised DuPont aca method and recommended IFCC method. AB - The method for continuous-flow assay of aspartate aminotransferase with the Technicon SMAC was modified to include preincubation of the serum enzyme with pyridoxal 5'-phosphate, to be consistent with the recommendations of IFCC and the Standards Committee of AACC. Preliminary estimates of the imprecision of the modified method on SMAC gave day-to-day standard deviations of 5.3 U/L at mean of 48 U/L (n = 66) and 6.2 U/L at 155 U/L (n = 61). Added bilirubin, sodium pyruvate, ascorbic acid, and endogenous lipids did not interfere. Comparison of results for 50 samples by this method with those by the manual IFCC method gave y = 1.1113x - 0.3 U/L, Sy/x = 4.4 U/L, and r = 0.997. Similar data are presented for the revised AST method for the DuPont aca discrete analyzer. Clinical data show that AST activities increase by as much as 200% when the serum is preincubated with pyridoxal 5'-phosphate. PMID- 7471431 TI - Adsorbent treatment and immunoaffinity chromatography compared for removing estradiol-17 beta from plasma. AB - Removal of estradiol-17 beta from plasma by immunoaffinity chromatography on anti estradiol antibodies coupled to Sepharose was investigated and compared with the use of charcoal adsorbent. Both methods completely removed added [3H]estradiol from pooled male plasma. In contrast to the adsorbent treatment, which caused marked changes in the biochemical composition of the plasma, immunoaffinity chromatography caused only minor perturbations. We discuss the advantages of using plasma, stripped of endogenous estradiol or other steroids by immunoaffinity chromatography, in construction of radioimmunoassay calibration curves. PMID- 7471433 TI - Method variation in lactate dehydrogenase isoenzyme determination. AB - Differences between methods for determining lactate dehydrogenase isoenzymes are illustrated, which can account for discrepant results. They should be taken into consideration in the interpretation of test results for the diagnosis of myocardial infarction. PMID- 7471434 TI - Radioenzymic measurement of norepinephrine, epinephrine, and dopamine: stability, enzymic activity, and sensitivity. AB - We investigated factors affecting sensitivity and reproducibility of radioenzymic (catechol-O-methyltransferase) measurement of plasma catecholamines. There was no apparent deterioration of catecholamine during storage at -20 degrees C for 240 days. Accuracy and sensitivity of the assay depended mostly on the method of transferase preparation; its preparation by adjusting the pH to 6.8 before fractionation with ammonium sulfate resulted in an unsuitable enzyme with a relatively high background-radioactivity count, particularly for neorepinephrine and dopamine. PMID- 7471435 TI - Method for rapidly estimating specific radioactivity and radiochemical purity of radioiodinated peptide hormones used in radioimmunoassays. PMID- 7471436 TI - Centrifugal enzymic analysis for lactate in plasma and cerebrospinal fluid. PMID- 7471437 TI - Ultracentrifugation does not affect results of analyses for digoxin, cortisol, and calcium in lipemic sera. PMID- 7471438 TI - Glucose phosphate isomerase and glutathione reductase in benign and malignant extrahepatic cholestasis. PMID- 7471439 TI - Colorimetry vs column chromatography in Hb A1 assay. PMID- 7471440 TI - Bence Jones proteinuria revisited. PMID- 7471441 TI - A Jendrassik-Grof method modified to eliminate hemoglobin interference with assay of total serum bilirubin. PMID- 7471442 TI - Elimination of a potential interference in assay for plasma caffeine. PMID- 7471443 TI - Advantages of use of ascorbic acid and sodium lauryl sulfate in the microdetermination of serum inorganic phosphorus. PMID- 7471444 TI - High values for lactate dehydrogenase in measles. PMID- 7471445 TI - Interference of hemoglobin H in the column-chromatographic assay of glycosylated hemoglobin A. PMID- 7471446 TI - Whole plasma or plasma water: the problem of the reference base. PMID- 7471447 TI - Iodination of antiferritin by the method of Bolton and Hunter. PMID- 7471448 TI - Semi-automated determination of non-esterified fatty acids in serum. PMID- 7471449 TI - Concerning a new pathogenetic hypothesis of the E.P.H. gestosis (note I). AB - The Authors, after having evaluated the literature data, report, on the basis of their clinical experience, a new pathogenic hypothesis of the E.P.H. gestosis which would be: "Disease due to missed materno-foetal adaptation:, correlated to the activation of the maternal immunocompetent system against the "not self" antigens, carried by the trophoblast and codified by the genoma of the father's origin. It is also due to missed destruction of the tunica media of the placental bed arteries and to the thromboplastinic activity of the trophoblastic cells damaged by the immunologic reaction. This disease, clinically represents, the appearance of a "temporaneous" (pre-eclampsia) or "definitive" (eclampsia) defeat of the fibrinolytic control mechanisms against a disseminated intravascular coagulopathy. PMID- 7471450 TI - Neurosurgical indications during pregnancy. AB - The Authors take into consideration some important forms of vascular and tumoral pathology of neurosurgical interest which can affect pregnant women. Some criteria of urgency or delay of the surgical treatment are proposed in the attempt of saving the mother's and the foetus' life. PMID- 7471451 TI - HGH behaviour at term of pregnancy and during puerperium. AB - Human growth hormone (HGH) has been measured in the plasma of 30 subjects at term of pregnancy, at 96 and at 144 h after delivery. The subjects were then selected into three groups: 10 were studied in basal conditions, 10 were given pyridoxine, 10 were given enantate testosterone and valerianate estradiol. In the first group the correlation index (t of Student) was not significant showing the lack of correlation among the tested averages. IN the second group the index of Student was weakly significant (t = 2.36, p less than 0.05). In the third group the Authors found a high representative correlation between the term of pregnancy and the 144th hour after delivery (t = 3.81, p less than 0.01) and between the 96th and 144th hour after delivery (t = 2.95, p less than 0.01). PMID- 7471452 TI - Evaluation of fetal pulmonary maturity by gas-chromatographic analysis of amniotic fluid palmitic acid/stearic acid (P/S) ratio. AB - Gas-chromatographic analysis of the fatty acids (P/S ratio) in 212 samples of amniotic fluid, 35 samples of vernix caseosa and 35 samples of the pellets obtained after centrifugation of the amniotic fluid were carried out to evaluate the effects of contaminants that might be present in amniotic fluid on the P/S ratio. The P/S ratio is used as an index of the degree of maturity of the fetal or neonatal lungs. Analysis of variance applied to the means of the P/S values for each of above materials showed them to be significantly different and attests to the importance of centrifugation for obtaining valid results. We propose a standard procedure of centrifugation for 60 minutes at 3500 X g followed by extraction and gas-chromatography as a rapid, valid way to measure the P/S ratio. PMID- 7471453 TI - Endometrial cytology during menstrual cycle, menopause and progestinic treatment. AB - The AA. examined 72 cytological smears of the human normal endometrium. Twenty three were taken during the proliferative phase, 17 during the secretory phase, 21 during menopause and 11 during estro-progestinic treatment. The study pointed out that the endometrial smear, which has been employed up to now almost exclusively in the cyto-oncological field, is also quite suitable for a dynamic cyto-functional evaluation of ovarian function, because of the series of interpretative elements, easily and readily identifiable at the microscope, which it furnishes. PMID- 7471455 TI - The validity of the urocytogram compared to colpocytology in hormonal cytodiagnosis. AB - The AA. studied the degree of significance in results obtained from a parallel urocytologic and colpocytologic study in sixty-two regularly menstruated women. The analysis of the data showed a significant correspondence between the results of both procedures regarding the karyopyknotic index, although the curve obtained by urocytogram was constantly lower than that one by colpocytology. From the study it emerges clearly the validity of urinary cytology for hormonal evaluation as alternative to colpocytology where the evaluation is often negatively influenced by many local factors. PMID- 7471454 TI - Ultrasound monitoring of gestosis polycentric study. AB - In polycentric research we studied 62 selected cases of gestosis to evaluate which symptom is greatly correlated with the deficit of fetal growth. According to us, the earlier the syndrome appearance the clearer the deficit of fetal growth. PMID- 7471456 TI - Amniotic fluid reverse triiodothyronine in normal and high risk pregnancies. AB - The authors have dosed by radioimmunoassay the rT3 concentrations in the amniotic fluid of 12 normal pregnant women and of 25 women with high risk pregnancies at the same period of gestation. The average of the rT3 concentrations in the amniotic fluid of pregnant women affected by diabetes mellitus, EPH gestosis and with foetal growth, retardation, are not different from those found in the control group. In the patients affected by Rh isoimmunization, the rT3 levels are remarkably higher than those found in the control group. The rT3 highest levels found in the amniotic fluid of pregnant women affected by Rh isoimmunization could be due. 1) to a decreased transformation of rT3 in rT2 for an accentuated alteration of the sulphydryl groups: 2) to a need of blocking the pathway's conversion of T4 in T3 (hormone with great catabolic effect) in these foetuses already presenting an accentuated catabolism for the chronic hemolytic enemy and for the severe hepatic alterations. PMID- 7471457 TI - Scanning electron microscopy of the human normal endometrium. AB - The AA. examined by scanning and transmission electron microscopy (SEM) the normal human endometrium of 21 females, 18 in fertile age and 3 in menopause. The study pointed out that the presence of cells with cilia and microvilli only in fertile women. This observation indicates the role these structures may have in the kinetic of spermatozoa, in the nidation and nutrition of oocyte and, at the same time, their dependence on the ovarian hormonal activity. PMID- 7471458 TI - Serum potassium in mother and fetus during normal delivery and cesarean section. AB - The influence of i.v. succinylcholine in term pregnant women on serum potassium of the fetus and of their own blood samples of two groups were taken to estimate the potassium level of women and their fetuses: 34 women delivering spontaneously and 24 by cesarean section using general endotracheal anesthesia. No significant differences were found in the newborns of serum potassium level of both groups, a finding which teaches, that succinylcholine given to the mother, has no significant influence on the serum potassium level of the newborn. Likewise, no significant change was foreseen in the serum potassium level in women after introduction of succinylcholine during cesarean section, in contrast to an increase of the serum potassium reported in the literature. PMID- 7471459 TI - Pitfalls in interpretation of the pulmonary valve echogram in patients with congenital heart disease. PMID- 7471460 TI - Primary pulmonary hypertension in adults; clinical features, catheterization findings and long-term follow up. PMID- 7471461 TI - Continuous ambulatory peritoneal dialysis. PMID- 7471462 TI - Acute gangrenous proctitis. PMID- 7471463 TI - Nelson's syndrome; a new look. PMID- 7471464 TI - Methotrexate in the treatment of rheumatoid arthritis; pilot study. PMID- 7471465 TI - Aortorenal vein graft stenosis causing recurrent hypertension; repair with a temporary shunt. PMID- 7471466 TI - Serial assessment of doxorubicin cardiotoxicity in pediatric patients with quantitative radionuclide angiocardiography. PMID- 7471467 TI - Sporotrichoid Mycobacterium marinum skin infection treated with minocycline hydrochloride. PMID- 7471468 TI - Intracellular and extracellular alpha-D-mannosidase activity of cultured skin fibroblasts: relationship to cystic fibrosis. AB - Fibroblast intracellular alpha-D-mannosidase was inactivated by 30--50% after 120 min at 50 degrees C. Separation of alpha-D-mannosidase by isoelectrofocusing and wheat germ lectin-Sepharose into neutral and acid components showed the former to be heat labile and the cause of the observed loss. Fetal calf serum acid alpha-D mannosidase could be inactivated at 60 degrees C if kept at pH 9.0. Fibroblast extracellular alpha-D-mannosidase was inactivated by 50--70% after 120 min at 50 degrees C, reflecting the greater level of the neutral component. Control and cystic fibrosis alpha-D-mannosidase showed no difference. PMID- 7471469 TI - A simple and reliable enzymatic assay for the determination of formic acid in urine. AB - A simple, reliable and efficient enzymatic assay for the analysis of formic acid in urine is described. The principle of the method is the quantitative oxidation of the acid with the enzyme formate dehydrogenase (FDH) in the presence of NAD. The detection limit was 0.5 mg/l, the recovery rates ranged from 66% to 74% and the coefficient of variation was 5%. The results obtained with this method correlated well with the values measured with two different gas chromatographic procedures. Normal formic acid excretion in urine is between 2.0 and 30.0 mg/l, with a mean of 13.0 +/- 7.4 mg/l and between 1.2 and 17.5 mg/g creatinine, the mean being 7.7 +/- 4.0 mg/g creatinine. The determination of formic acid concentration in urine may be an important means to quantify the degree of exposure to or intoxication of persons with methanol, formaldehyde and acetone. PMID- 7471470 TI - Advantages of CUSUM techniques for quality control in clinical chemistry. AB - The performance of routine analytical laboratories is assessed, inter alia, by the use of appropriate internal quality control techniques. Despite evidence that the cumulative sum technique is generally superior to Shewhart-type control charts and many others, its use has been limited by the popular misconception that it is inferior to Shewhart's in detecting large variations and/or outliers in quality control data. The application of computer simulation methods has enabled us to answer this criticism and has provided the basis for further improvements in the design of the appropriate control scheme for general use in clinical chemistry. PMID- 7471471 TI - No relationship between lactate dehydrogenase and estrogen receptor levels in clinical human breast cancer. AB - The level of lactate dehydrogenase has been reported to increase in a human breast cancer cell line as a result of estrogen action [4]. This finding suggests that lactate dehydrogenase may be useful in clinical human breast cancer as a marker for intactness of the estrogen receptor pathway and for probability of response to hormonal therapy. In 97 hunan breast cancer specimens submitted for estrogen receptor analysis, the level of lactate dehydrogenase varied from 0.19 to 3.3 I.U. per mg of extractable protein and bore no relation with either the presence of estrogen receptor or its level. It thus appears that lactate dehydrogenase is not a useful marker for endocrine responsiveness in clinical breast tumors. PMID- 7471472 TI - Improved radioimmunoassay for human TSH. AB - This study concerns the optimization of the human TSH (h-TSH) radioimmunoassay with special emphasis on reducing the heterogeneity of the 125I h-TSH tracer. Enzymatic iodination of h-TSH with glucose oxidase/lactoperoxidase was shown to be superior to either low or high dose chloramine-T procedures, producing a high specific activity reagent (70--150 microCi/microgram) with minimal evidence of damage. Tracer purification procedures not only affected initial immunoactivity but also storage stability and heterogeneity of the resulting 125I h-TSH. Tracers purified by combining concanavalin A-Sepharose adsorption and high resolution gel filtration (Sephadex G100), produced significantly lower (p less than 0.001) serum h-TSH measurements than were observed in less purified tracer materials. Concanavalin A-Sepharose adsorption yield of the 125I h-TSH iodination products closely correlated with the yield (r = 0.85, p less than 0.001) and immunoactivity (r = 0.90, p less than 0.001) of the tracer produced, thus making this an ideal method for initial tracer purification. Storage of tracer adsorbed to a solid support (concanavalin A-Sepharose) reduced technical manipulations without compromising tracer performance. Loss of specific activity was minimized by storage at -70 degrees C. The assay developed using these technical approaches showed a sensitivity limit of 0.005 +/- 0.001 (S.E.M.) microU/tube; 50% displacement at 0.18 +/- 0.08 (S.E.M.) microU/tube and complete delineation between euthyroid (n = 49, 2.44 +/- 0.18 (S.E.M.) mU/l, range 1.00--6.08) and hyperthyroid (n = 62, 0.34 +/- 0.02 (S.E.M.) mU/l, range 0.10--0.85), serum h-TSH levels. PMID- 7471474 TI - The relationship between unbound and total cortisol: its usefulness in detecting CBG abnormalities. AB - The relationship between unbound and total cortisol has been studied in patients with a variety of clinical conditions. We report on a simple, rapid and reliable method for evaluating the percentage unbound cortisol in serum or plasma which can be used in conjunction with total plasma cortisol to obtain a more complete understanding of the patients' adrenal status. Comparison of a patient's percentage unbound and total cortisol with a nomogram showing the normal relationship between percentage unbound and total cortisol indicates the patient's concentration of cortisol binding globulin. The concentration of unbound cortisol, the biologically active moiety, is the product of the percentage unbound and total cortisol concentration. The following values for the unbound cortisol concentration (microgram/dl) were obtained (mean +/- S.D.). Twenty-four normal subjects, 8--10 a.m., 1.2 +/- 0.4; 14 women receiving an oral contraceptive, 1.4 +/- 0.5; 6 patients with adrenal insufficiency, 0.2 +/- 0.1; 9 hyperthyroid patients, 1.7 +/- 0.8; 5 acute ill patients, 3.8 +/- 2.4; and a patient with Cushing's syndrome, 6.1. In normal subjects the values decreased at 4 p.m. (0.4 +/- 0.1) and after dexamethasone administration (0.1 +/- 0.1), and increased following the intravenous injection of adrenocorticotropic hormone (3.6 +/- 0.7). In pregnant women the unbound cortisol increased as the pregnancy progressed, first trimester: 1.2 +/- 0.3, second trimester: 1.6 +/- 0.2, third trimester: 2.4 +/- 0.5. PMID- 7471473 TI - Connective tissue activation. XVII. Radioimmunoassay of a human platelet derived connective tissue activating peptide (CTAP-III) and specificities of anti-CTAP III sera. AB - The platelet-derived connective tissue activating peptide (CTAP-III) has been shown to be an important factor stimulating the metabolism and proliferation of human connective tissue cell strains, including synovial tissue cells. The quantities of CTAP-III affecting the cellular changes and the amounts of various biologic fluids and tissues are small. The objectives of this study were to develop a radioimmunoassay (RIA) for CTAP-III and to ascertain the specificities of the anti-CTAP-III sera reagents. The antisera were shown not to cross-react with a number of polypeptide hormones. However, two other platelet proteins, beta thromboglobulin and low affinity platelet factor-4, competed equally as well as CTAP-III for anti-CTAP-III antibodies in the RIA system. Thus, the three platelet proteins are similar or identical with respect to those portions of the molecules constituting the reactive antigenic determinants. The levels of material in normal human platelet-free plasma that inhibited anti-CTAP-III--125I-CTAP-III complex formation were determined to be 34 +/- 13 (S.D.) ng/ml. PMID- 7471475 TI - Erythrocyte porphyrinogen carboxy-lyase activity in porphyria cutanea tarda and certain other human porphyrias. AB - Red cell porphyrinogen carboxy-lyase activity was measured using uroporphyrinogen III as substrate in 18 normal persons, 7 male patients with porphyria cutanea tarda, 3 female patients with erythropoietic protoporphyria and 2 female patients with variegate porphyria. The mean values obtained in normal subjects were 0.151 nmol of uroporphyrinogen disappeared in 30 min per mg of protein, and 0.038 nmol of coproporphyrinogen formed in 30 min per mg of protein. We have not been able to detect significant differences between males and females. In porphyria cutanea tarda the enzyme activity was the same as in normal subjects considering either substrate disappearance or end product formation. The differences were not significant at the p less than 0.05 level. Patients with variegata porphyria also exhibited normal erythrocyte porphyrinogen carboxy-lyase activity. The enzyme activity of erythrocytes from patients with erythropoietic protoporphyria was higher than in normals; mean values for specific activities being 0.204 nmol of uroporphyrinogen disappeared, and 0.071 nmol of coproporphyrinogen formed. The significance of the results with respect to the chemical picture of different porphyrias is discussed. PMID- 7471476 TI - Serum IgM in primary biliary cirrhosis. AB - Serum IgM concentrations were measured in 25 patients with primary biliary cirrhosis (PBC) and 25 age- and sex-matched controls by the classical Mancini technique and by a modified method, which included reduction to 7 S monomers. Standards calibrated against the WHO standard, as well as a serum standard with an absolute value calibrated by immunological and non-immunological techniques, were used. All patients had an elevated IgM level; measurement of serum IgM is therefore a simple and sensitive screening test for patients with cholestasis. When measured with the standard calibrated against the WHO standard, the average IgM levels for PBC patients and controls agreed with results previously reported; the average IgM levels were much lower when a serum standard with an absolute IgM value was used; further standardization in needed. The differences in the ratio of the IgM measured by the classical method (pentameric IgM) to that measured by the alternative method (monomeric IgM) support the existence of different IgM subgroups or the in vivo presence of monomeric IgM in some patients with PBC. PMID- 7471477 TI - Altered erythrocyte spectrin extractability in Duchenne muscular dystrophy. AB - Spectrin extractability was measured in the erythrocyte membranes obtained from patients with Duchenne muscular dystrophy (DMD) and normal matched controls. After the extraction of spectrin by adding deionized water, an aliquot of the pellet was applied to 6% SDS-polyacrylamide gels. Spectrin-Band 3 ratio on the gels was used as an indicator of spectrin extractability. Our data disclosed that spectrin extractability is significantly reduced in DMD patients when compared with that of controls and these results suggest that spectrin from DMD patients is more tightly bound to the membranes. Our data may also be applicable to the discussion of the relationship of spectrin to other membrane properties as well as to previously described membrane abnormalities in DMD erythrocytes. PMID- 7471478 TI - Prenatal detection of defects in propionate metabolism. PMID- 7471479 TI - Rapid and efficient extraction of steroids from microlitre quantities of plasma using Extrelut mini-columns. PMID- 7471480 TI - [Determination of erythrocyte transketolase activity (author's transl)]. PMID- 7471481 TI - Assessment of a simple electrophoretic method for measuring HbA. PMID- 7471482 TI - Enzymatic assay of serum sialic acid. AB - Serum sialic acid has been assayed enzymatically. The reaction includes neuraminidase hydrolysis of glycoprotein, cleavage of sialic acid to pyruvate by N-acetyl neuraminic acid (NANA)-aldolase, oxidation of pyruvate by pyruvate oxidase which produces hydrogen peroxide, and colorimetry of hydrogen peroxide using the peroxidase-p-chlorophenol-4-amino-antipyrine method. This method showed good correlation between a chemical method (r = 0.984), good recovery (98.8%) and good reproducibility (within-run-precision: 1.0% C.V.; day-to-day precision: 1.9% C.V). Intrinsic serum pyruvate produces an equimolar positive effect. The normal value range is 1.94 +/- 0.29 mmol/l (mean +/- S.D.,n = 24). PMID- 7471483 TI - International Federation of Clinical Chemistry scientific committee expert panel on pH and blood gases (EPpH) and International Union of Pure and Applied Chemistry division of clinical chemistry commission on quantities and units in clinical chemistry (CQUCC). Document stage 2, draft 1 (1979-10-10) with a view to a recommendation. Physico-chemical quantities and units in clinical chemistry with special emphasis on activities and activity coefficients. PMID- 7471485 TI - International Federation of Clinical chemistry. Approved recommendation (1979) on quality control in clinical chemistry. PMID- 7471484 TI - Preparation of antibodies to catecholamines and metabolites--syntheses of various immunogens and characterization of the resulting antibodies. AB - The synthesis of antigens of the haptens p-tyramine, octopamine, synephrine, dopamine, normetanephrine, beta-(3,4-dichlorophenyl)-ethylamine and 3-fluoro tyramine is described. The coupling of the haptens to the carrier protein was performed either via the side chain by means of succinic anhydride, or through the phenolic ring via an aminomethyl group which was introduced by a Mannich reaction. Antisera to these haptens were produced in rabbits by immunization with these antigens. The antisera were characterized by determination of titer and specificity (p-tyramine, synephrine, normetanephrine). The possibility is discussed of using antisera to p-hydroxyphenylethylamine for a radioimmunological determination of catecholamines. PMID- 7471486 TI - Complications in the determination of HDL2/HDL3 ratios. AB - We investigated different procedures to subfractionate high density lipoprotein (HDL) into its density subfractions (HDL2 and HDL3). When pore-limit as well as zone polyacrylamide gel electrophoresis was used, it proved to be necessary to use freshly prepared and plasma protein-free HDL preparations. For a quantitative subractionation of HDL in HDL2 and HDL3 we also tested density gradient ultracentrifugation. To avoid interference with LDL contamination a low starting density was required. Both methods, electrophoresis as well as density gradient ultracentrifugation were too laborious and time-consuming for large scale use. We devised a combination of both ultracentrifugation and electrophoresis as a method which is relatively appropriate for determination of HDL2/HDL3 ratios on large scale. PMID- 7471487 TI - [Erythrocyte bilirubin: practical implications of variations in concentration (author's transl)]. AB - The values of red blood cell bilirubin and unconjugated plasma bilirubin have been measured simultaneously in 31 blood samples of 21 new-born infants with deep jaundice. The methods of measurement used by the authors have proved quite reliable and there is good correlation between both values. The concentrations rise together with the bilirubin/albumin molecular ratio. Nevertheless even with a low ratio, the red blood cell bilirubin rises to high levels when acidosis is present. PMID- 7471488 TI - A case of an exceptionally high level of urinary-4-hydroxyproline containing polypeptides. AB - The determination of total urinary-4-hydroxyproline in an adolescent showed a very high excretion of hydroxyproline containing peptides: 26.2 mmol/24 h. Estimation of total urinary 3-hydroxyproline and peptide fractionation on Biogel P 2 demonstrated unusual features. Such cases are of great interest for structural studies of the urinary polypeptides and understanding of collagen catabolism. PMID- 7471489 TI - The use of concanavalin A crossed immuno-affinoelectrophoresis to detect hormone associated variations in alpha 1-acid glycoprotein. AB - alpha 1-Acid glycoprotein produces three peaks on crossed immuno affinoelectrophoresis with concanavalin A in the first dimension, which is indicative of a degree of heterogeneity in the carbohydrate portion of this plasma glycoprotein. A different pattern, however, showing relatively less concanavalin A-binding material, is found in association with and after an increase in female sex hormone levels. Our observations strongly suggest a prolonged hormonal effect on the carbohydrate composition of alpha 1-acid glycoprotein, which may alter the metabolism or the function of the protein. PMID- 7471490 TI - Vitamin B12-dependent methyltetrahydrofolate: homocysteine methyltransferase activity in normal and leukemic human hematopoietic cells. AB - The cyanogen bromide method was applied to the assay of vitamin B12-dependent methyltetrahydrofolate:homocysteine methyltransferase activity in normal and leukemic human hematopoietic cells. Normal peripheral lymphocytes and leukemia cells of lymphoid origin wuch as CLL and ALL, contained higher levels of enzyme activity than did normal human bone marrow cells. Normal granulocytes and leukemia cells of myeloid origin, such as CML in the chronic phase and AML, contained lower enzyme activity. Leukemia cells of CML in blast crisis showed higher mean activity than in the chronic phase of the disease. PMID- 7471491 TI - Measurement of gamma-aminobutyric acid in human cerebrospinal fluid: radioreceptor assay using [3H]muscimol. AB - A method is described for the determination of gamma-aminobutyric acid (GABA) levels in human cerebrospinal fluid by modification of the radioreceptor assay utilizing [3H]muscimol as labelled ligand. This method is compared with the radioreceptor assay using [3H]GABA as labelled ligand. Although the [3H]muscimol assay is less sensitive than the [3H]GABA method, it offers the advantage of being more rapid due to the use of a filtration step instead of the usual, more time-consuming centrifugation of the samples. Samples of CSF of patients with various neurological or psychiatric disturbances were analysed. There was a satisfactory correlation between the GABA values obtained by the two assays. PMID- 7471492 TI - Nuclear androgen receptors in human prostatic tissue. Extraction with heparin and estimation of the number of binding sites with different methods. AB - A procedure for the estimation of nuclear androgen receptors in benign prostatic hyperplastic tissue is described, which employs extraction of receptors from nuclei with buffers containing heparin. Extraction of a nuclear pellet with a heparin-containing (1 g/l) buffer appeared to have definite advantages over 0.4 mol/l KCl extraction. Heparin appeared to be twice as efficient in extracting androgen receptors. In addition aggregated receptor proteins, formed after storage at -80 degrees C, were partly deaggregated by heparin. Specific isolation of the androgen receptor was performed using either agar gel electrophoresis, protamine sulphate precipitation or LH-20 gel filtration. A comparison was made between the amounts of estimated receptors with these different techniques. Protamine sulphate precipitation resulted in the highest estimates of receptor bound 5 alpha-[3H]dihydrotestosterone (3H-DHT). Treatment of the labelled nuclear extracts with a charcoal suspension prior to the receptor assay resulted in lower amounts of estimated androgen receptors. A method for routine evaluation of nuclear androgen receptors in prostatic tissue has been evaluated, which involves extraction of nuclear pellets with a heparin-containing (1 g/l) buffer, exchange labelling of the nuclear extracts for 20 h at 10 degrees C and quantification of the receptors with protamine sulphate precipitation. PMID- 7471493 TI - Normal excretion of m-hydroxymandelic acid in hypertensive patients. AB - o-Hydroxymandelic acid (OHMA), m-hydroxymandelic acid (MHMA) and p hydroxymandelic acid (PHMA) were measured in the urine of 42 normotensive and 54 hypertensive patients. Patients having high urinary MHMA levels were all found to be ingesting medications containing m-synephrine (phenylephrine). These patients also had high levels of urinary m-synephrine which was excreted as the glucuronide. When patients ingesting m-synephrine were excluded from the analysis, no significant differences were observed between the two groups for the urinary excretion of OHMA, MHMA and PHMA. PMID- 7471494 TI - Fluorometric assay of B6 vitamers in biological material. AB - A method for the separation and assay of all B6 vitamers from a single sample of serum is reported. A new reagent system containing sodium glyoxalate, potassium aluminum, sulfate and manganese dioxide was used for quantitative conversion of pyridoxamine and its phosphate to the corresponding aldehydes by a combination of transamination and oxidation reactions. PMID- 7471496 TI - Porphyrin-induced photodamage at the cellular and the subcellular level as related to the solubility of the porphyrin. AB - Porphyrin-induced photodamage has been studied on small organic molecules, biomolecules, mitochondria and red cells. Water soluble components (e.g. tryptophan and glutamate dehydrogenase) are more easily destroyed by uroporphyrin than by protoporphyrin. On the other hand, lipophilic components (e.g. succinate dehydrogenase, mitochondria and red cell membranes) are more severely damaged by protoporphyrin. The results may be of importance to explain the different skin lesions in erythropoietic protoporphyria and in porphyria cutanea tarda. The photodamage is enhanced by D2O and reduced by azide. Reagents known to increase or decrease the yields of superoxide, peroxide or hydroxyl radicals have no effect on the photodamage. The results suggest that singlet oxygen is the most important reactive oxygen species. PMID- 7471495 TI - On the validity of a levo-norgestrel radioimmunoassay. AB - The validity of a levo-norgestrel (L-NOG) radioimmunoassay in human plasma was assessed using a test of radiochemical purity. Extraction with diethyl ether followed by chromatography on celite gave an L-NOG peak of constant specific activity, indicating the radiochemical purity of this compound. This was demonstrated with three different plasma pools. Hence, the chromatographic method was considered to yield valid results. When diethyl ether or petroleum ether extracts were assayed without chromatography, overestimates of 37% and 18%, respectively, were encountered. PMID- 7471497 TI - Urinary sialic acid levels in aspartylglycosaminuria. AB - Urinary sialoglycoconjugates were studied in 22 patients with inherited deficiency of 1-aspartamido-beta-N-acetylglucosamine amidohydrolase (aspartylglycosaminuria), in eight obligate heterozygotes, and in age- and sex matched control subjects. Total sialic acid excretion was significantly higher in the patients (38.3 +/- 17.7 mumol/mmol creatinine, mean +/- S.D.) than in the matched controls (17.7 +/- 7.3 mumol/mmol creatinine, p less than 0.001). The sialic acid output in the heterozygotes did not differ from that of the controls. Gel filtration studies revealed that the increase in urinary sialic acid in aspartylglycosaminuria is of bound type and confined to the low molecular mass region. A linear positive correlation was found between the output of sialic acid and glycoasparagine in the individual patients (r = 0.77, p less than 0.001). The amount of sialylated metabolites excreted in urine did not correlate with the severity of clinical manifestations in aspartyl-glycosaminuria. PMID- 7471498 TI - Abnormalities in folate binding pattern of serum from a patient with megaloblastic anemia and folate deficiency. PMID- 7471499 TI - NADH-ferricyanide reductase, a convenient approach to the evaluation of NADH methaemoglobin reductase in human erythrocytes. PMID- 7471500 TI - Sampling errors in the determination of urine calcium and oxalate: solubility of calcium oxalate in HCl-urine mixtures. PMID- 7471501 TI - Echoes from the cochlea. PMID- 7471502 TI - The fractured nose. PMID- 7471503 TI - Successful methotrexate therapy in oral florid papillomatosis. AB - Oral florid papillomatosis is a rare syndrome, which was first described under this name by Rock & Fisher in 1960. The same syndrome had been previously described as papillomatosis mucosae oris carcinoides by Scheicher-Gottron (1958). It is a benign condition, capable of malignant degeneration. Surgical excision and electrocoagulation are the most widely used therapeutic methods, despite the fact that relapse of symptoms after treatment by these methods is common. The literature comprises a few reports on favourable therapeutic effects obtained with small doses of methotrexate. This paper describes a patient with multifocal, very extensive form of oral florid papillomatosis, in whom both surgical excision and electrocoagulation were regarded as inadequate in view of the extensiveness of the lesion. Chemotherapy with methotrexate proved to be effective and resulted in complete remission. No relapse has occurred during a follow-up period of over 2 years. The value of chemotherapy with methotrexate in the treatment of an extensive form of oral florid papillomatosis is emphasized. PMID- 7471505 TI - Do perilymph fistulae cause otitis media--or vice versa? PMID- 7471506 TI - Muscular anomalies caused by delayed development in human aneuploidy. AB - Comprehensive dissections of three infants with trisomy-13 and three with trisomy 18 have revealed several muscles which appear to be "abnormal" at the time of birth and shortly thereafter. Careful observations show that the peculiar morphology of these muscles results from delayed development rather than from anatomical malformation. The observations are compared with physiological, pathological and in vitro experimental studies of delayed embryonic development in human aneuploidy. PMID- 7471507 TI - Athelia in a female infant - heterozygous for anhidrotic ectodermal dysplasia. AB - The case of a female infant with athelia is reported. Her mother, maternal aunt and grandmother show hypodontia, sparse hair and small breasts associated with mammillary hypoplasia. The clinical features and the results of MINOR's sweat test suggest a heterozygous state of anhidrotic ectodermal dysplasia as the most likely explanation. PMID- 7471504 TI - Eighteen-year report on stapedectomy. III. Postoperative complications. AB - This third part of our series develops the problems of postoperative complications with their various solutions. It also deals with the postoperative progressive cochlear deterioration and its origins. PMID- 7471508 TI - Partial trisomy 6q, due to balanced maternal translocation (6;22) (q21; p13) or (q21; pter). AB - We report a stillborn infant with partial trisomy 6q who had several major congenital malformations not previously associated with the chromosomal aberration. These included occipital encephalocele, ambiguous genitalia with imperforate anus, omphalocele and unilateral hydronephrosis. The infant's karyotype was 46,XY,-22,der(22),t(6;22)(q21; p13) or (q21;pter)mat. The mother and maternal grandmother are balanced translocation carriers. PMID- 7471509 TI - Monosomy 10qter due to a balanced familial translocation: t(10;16)(q25.2;q24). PMID- 7471510 TI - No "fragile X" chromosome in normal men. PMID- 7471511 TI - A recognizable phenotype in a child with partial duplication 13q in a family with t(10q;13q). AB - A case of partial duplication 13q14 leads to qter is reported in a 9-year-old male with clinical symptoms which include trigonocephaly and synophrys, producing an easily identifiable phenotype. The chromosome duplication resulted from a familial t(10;13)(qtr;q14). Subsequently, a normal balanced carrier sibling was diagnosed prenatally. PMID- 7471512 TI - 49,XYYYY. A case report. AB - A 14-month-old boy with a 49,XYYYY karyotype is reported. The physical examination revealed unusual facial features, brachydactyly with clinodactyly, limitation of supination at the left elbow, and inguinal hernia. Radiological abnormalities of the skeleton and urinary tract were present, and the developmental examination showed a DQ of 70. PMID- 7471513 TI - Clinical and neurophysiological findings in heterozygotes for nonketotic hyperglycinemia. AB - Heterozygotes for nonketotic hyperglycinemia (NKH), a disorder of glycine degradation, have a slightly abnormal metabolism of glycine. As the severe neurological symptoms which are characteristic for the homozygotes are at least partially due to a disturbance of glycine function as a neurotransmitter, minor neuronal dysfunctions might be expected also in heterozygotes. Although their general health was within normal limits in these 13 heterozygotes, slight neurological symptoms and signs were observed. Neurophysiological investigations revealed disturbance of the vestibular function in six subjects, preponderance of beta-wave activity in five, subnormal amplitude of a-wave, and shortening of implicit time of the first oscillatory potential (OPI) in the retinography. Functioning of peripheral nerves appeared normal in measurements of motor conduction velocity, distal latency and amplitude of muscle response. These minor dysfunctions of the central nervous system may well be due to a slightly abnormal degradation of glycine in heterozygotes for nonketotic hyperglycinemia. PMID- 7471515 TI - Lymphocyte blastogenesis in nephrotic syndrome. AB - Peripheral blood lymphocytes from children with steroid-responsive nephrotic syndrome showed inhibition of the blastogenic response to PHA-P when incubated in the presence of autologous serum. Inhibition was also evident when patients' cells were incubated with normal human serum or when normal cells were incubated with patients' serum. This inhibition was not seen when the patients were in remission. In experimental nephrosis in rats induced by a single injection of the aminonucleoside of puromycin, a similar pattern of lymphoblastogenic inhibition was seen. Thus human steroid-responsive nephrotic syndrome and experimental nephrosis both show similar disturbances of lymphocyte responsiveness to mitogenic stimulation. PMID- 7471514 TI - Circulating antigens, immune complexes and C3d levels in human schistosomiasis: relationship with Schistosoma mansoni egg output. AB - Circulating schistosome antigens (CSA), circulating immune complexes (CIC) and C3 breakdown product - C3d - were investigated in human schistosomiasis in comparison to the S. mansoni egg count. A close relationship was observed between the mean number of eggs/g of stool and the detection of CSA (evaluated by the radioimmunoprecipitation-PEG assay - Ripega), CIC (Clq-binding test) and C3d levels (quantitated by radial immunodiffusion). All the patients with more than 500 S. mansoni eggs/g of stool also presented antigen '4', specific of the genus Schistosoma, in the serum. A significant correlation was noticed between levels of CSA and CIC. This suggests the involvement of several schistosome antigens in the detected CIC. No relationship was noted between CIC and C3d levels. In contrast, there was a highly significant correlation between levels of CSA and C3d. The interaction between certain schistosome antigens and the complement system is discussed. PMID- 7471516 TI - PPD-induced viral antibody production in human blood lymphocytes. AB - The ability of purified protein derivative of tuberculin (PPD) to induce polyclonal antibody production in cultures of human blood lymphocytes was studied. IgG and IgM were determined by the enzyme-linked immunosorbent assay (Elisa). PPD induced both IgM and IgG production, with a predominance of IgM. PPD usually stimulated a stronger IgM response but a weaker IgG response than did pokeweed mitogen (PWM). Supernatants of PPD- or PWM-stimulated lymphocyte cultures were tested for antibodies to morbilli, rubella and herpes simplex by Elisa. PPD as well as PWM induced viral antibody production in lymphocytes of donors who had serum antibodies to the corresponding viral antigens. Production of viral antibodies of IgG class but not of IgM class was demonstrated. The PPD induced antibody response was T cell-dependent. PMID- 7471518 TI - Agonist interaction and decentralization supersensitivity in the nictitating membrane of the cat. AB - 1. The responses to individual drugs and to combinations of two drugs were studied in normal and chronically decentralized nictitating membranes (NM) of cats. Noradrenaline (NA), acetylcholine (ACh), serotonin (5HT) and potassium chloride (KCl) and the combinations NA + ACh, NA + 5HT, NA + KCl and ACh + 5HT were injected intra-arterially towards the smooth muscle. 2. Chronic decentralization resulted in supersensitivity to each of these agonists when injected alone. 3. The contraction of the normal NM caused by each drug combination showed potentiation in comparison to the sum of contractions caused by the same drugs given separately. The same drug combinations showed no potentiation in decentralized muscle. 4. Decentralization supersensitivity and potentiation due to agonist interactions in normal preparations may share a common mechanism. The involvement of calcium in this phenomenon is discussed. PMID- 7471519 TI - Adrenaline depletion in the hypothalamus of the rat after chronic alpha methyldopa. AB - 1. Individual anterior hypothalamic-preoptic nuclei were dissected from the brains of control and alpha-methyldopa treated (2 x 40 mg/kg, 5 days) rats, and the concentrations of adrenaline and other catecholamines were estimated. 2. By a combination of radioenzymatic assay and paper chromatography the concentrations of adrenaline, noradrenaline, dopamine, alpha-methylnoradrenaline and alpha methyldopamine were determined concurrently in the same sample. 3. alpha Methyldopa reduced the adrenaline levels of the hypothalamic nuclei by 47--68% of control concentrations. 4. A differential displacement of the parent catecholamines was observed and the depletions were ranked: adrenaline less than dopamine less than noradrenaline. 5. alpha-Methylnoradrenaline and alpha methyldopamine were found in all hypothalamic nuclei. 6. The depletion of hypothalamic adrenaline by alpha-methyldopa could represent a pharmacological mechanism contributing to the antihypertensive action of the drug. PMID- 7471520 TI - In-vivo modification of angiotensin II pressor responsiveness in sheep by indomethacin. AB - 1. Sodium (Na) depletion, due to uncompensated loss of parotid saliva, reduces the pressor responsiveness to angiotensin II (AII) in intact conscious sheep. 2. Inhibition of prostaglandin synthesis by indomethacin restored AII pressor responsiveness in Na-deplete sheep. 3. Indomethacin infusion also increased the pressor responsiveness to AII in the Na-replete sheep. 4. These findings suggest that in vivo prostaglandins may play a role in modulating the pressor responsiveness to angiotensin II in sheep. PMID- 7471517 TI - The separation of human palatine tonsillar cells in the reorienting gradient zonal rotor: a large-capacity enrichment of plasma cells. AB - Velocity sedimentation by centrifugation in a previously described isokinetic gradient, at unit gravity, or with an elutriator has proved to be highly effective as a method for the enrichment of many kinds of cells. For many biochemical and immunological purposes, the available techniques for the separation of cells have been incapable of separating a sufficient number of cells. With a newly designed rotor, we have separated more than twenty-fold more human tonsillar cells that can be separated in the previously described isokinetic gradient. Without exceeding the band capacity, we separated 692 million tonsillar cells. Plasma cells were more than seven-fold enriched from human tonsillar cells. When 692 million tonsillar cells were separated in the rotor, the five most enriched, contiguous fractions of plasma cells contained 44 million cells. The most enriched fraction contained 27% plasma cells. We are not aware of any method for velocity sedimentation which will separate such large numbers of tonsillar cells. PMID- 7471521 TI - Effect of adrenocortical steroids and their structural analogues on blood pressure in sheep. AB - 1. A model of adrenocortical steroid-induced hypertension based on the effects of ACTH administration has been developed in sheep. The present studies examine the effects of a number of different steroid hormones on blood pressure to investigate their structure-activity relationships. 2. Infusion of the major ovine adrenal steroid hormones (combined steroid infusion of cortisol, corticosterone, 11-deoxycortisol, aldosterone, deoxycorticosterone) reproduced the metabolic but not the blood pressure effects of ACTH. 3. Addition of 17 alpha, 20 alpha-dihydroxyprogesterone and 17 alpha-hydroxyprogesterone, at rates appropriate for conditions of ACTH stimulation, to the combined steroid infusion reproduced both the blood pressure and metabolic effects of ACTH. 4. 17 alpha-20 beta-Dihydroxyprogesterone, 20 beta-dihydroxy-11-deoxycortisol and 20 beta hydroxycortisol all had additional hypertensive activity when given with combined steroid infusion, but 16 alpha-hydroxyprogesterone, 11 beta, 17 alpha dihydroxyprogesterone, 20 alpha-hydroxyprogesterone and 20 beta hydroxyprogesterone did not. 5. These studies support the concept of a new class of steroid hormone action in blood pressure regulation. PMID- 7471522 TI - Effect of depletion of spinal noradrenaline by 6-hydroxydopamine on the development of renal hypertension in rats. AB - 1. A single injection in rats of 250 microgram of 6-hydroxydopamine HCl (6-OHDA) into the subarachnoidal space of the spinal cord of rats resulted in a lasting, selective depletion of spinal noradrenaline. Dopamine levels in the spinal cord and catecholamine levels in various brain regions were not markedly affected. 2. When ether anesthesia was used spinal noradrenaline was found to be almost completely depleted by the administration of 6-OHDA. Only partial depletion was achieved when pentobarbitone anaesthesia or neuroleptic analgesia was used. 3. The blood pressure rise caused by electrical stimulation of the posterior hypothalamus was not affected by 6-OHDA treatment 7 days previously. 4. 6-OHDA administration did not influence the development of two-kidney Goldblatt hypertension. When 6-OHDA was administered 7 days before clipping, a slight delay of the development was observed, but this did not occur when 6-OHDA treatment was given 3--4 h before clipping. 5. It is concluded that intact spinal noradrenergic neurotransmission is neither a prerequisite for the development of two-kidney Goldblatt hypertension, nor for the pressor response to hypothalamic stimulation. PMID- 7471523 TI - Altered cardiac noradrenaline stores in DOCA-salt hypertensive rats. AB - 1. The storage of noradrenaline (NA) in hearts of DOCA-salt hypertensive rats was examined after in vivo administration of labelled NA. 2. The cardiac NA concentration and the neuronal NA storage capacity were reduced in these rats compared with normotensive controls. The extent of reduction was dependent on the duration of hypertension. 3. These findings are similar to observations on spontaneously hypertensive rats and support the hypothesis that storage of the cardiac sympathetic transmitter is impaired in DOCA-salt hypertension. PMID- 7471524 TI - Effects of spironolactone on serum bile acids. PMID- 7471525 TI - Effect of complement activation on granulocyte adherence to nylon fibers: inhibitory activity of C567 complex. PMID- 7471526 TI - Immunoregulatory abnormalities in familial Mediterranean fever. PMID- 7471527 TI - The artificial granuloma. III. Collagen synthesis during the cell-mediated granulomatous response as determined in explanted granulomas. PMID- 7471528 TI - Test for the effects of diagnostic levels of ultrasound on the immune response of mice. PMID- 7471530 TI - Antibodies to DNA in patients undergoing long-term hemodialysis. AB - It has been suggested that antibodies to native DNA occur with some frequency in patients treated by hemodialysis. This has been attributed to the use of the technique. We have investigated 48 patients on maintenance dialysis for the presence of antibodies to single stranded DNA (ss-DNA) and native DNA (n-DNA) compared to a control group of 25 nondialyzed patients with renal disease. In neither group of patients were antibodies to ss-DNA detected frequently. Antibodies to n-DNA were present in three of the dialyzed patients and in none of the control group. The frequency of antibodies in dialysis patients was insignificant in comparison to the same control group of non-dialyzed renal patients. There was no correlation between duration of dialysis and development of antibodies. A drug-induced lupus state is possible, but not established for two of the three patients on dialysis. PMID- 7471529 TI - Acute interstitial nephritis: correlations between clinical and morphological findings. AB - The clinical and morphological findings in 30 cases of acute interstitial nephritis were compared in an attempt to find possible correlations. Patchy and/or incompletely diffuse interstitial infiltration was considered a milder form of the disease, the prognosis of which was significantly better than that of diffuse disease (P less than 0.01, Wilcoxon test). The prognosis was significantly poorer when acute renal failure persisted for three weeks or more (P less than 0.01), Wilcoxon test). Serum creatinine values served as a criterion for evaluating the course and prognosis of the disease. The patients were assigned to one of four prognosis groups. PMID- 7471531 TI - Comparison of hemodynamics induced by conventional acetate hemodialysis, bicarbonate hemodialysis and ultrafiltration. AB - Six patients on conventional acetate hemodialysis who had a marked decrease in total peripheral resistance index (TPRI) and hypotension after dialysis were selected for study. Systemic hemodynamic factors such as cardiac output, blood pressure, hematocrit and heart rate were measured, and the values were compared to those found after bicarbonate hemodialysis (Bi-HD) or ultrafiltration without simultaneous dialysis (UF) in the same patients. After Bi-HD and UF, the hemodynamic indices remained stable. Hematocrit level was significantly elevated after Bi-HD (+2.7%, P less than 0.05). Heart rate was insignificantly increased after each procedure. We concluded that Bi-HD can prevent unpleasant side effects induced by the decrease in TPRI with acetate hemodialysis. The stability of TPRI resulting from the increase in hematocrit level during Bi-HD may contribute to the prevention of cardiovascular instability. PMID- 7471532 TI - Acquired renal cysts in uremic patients--in vivo demonstration by computed tomography. AB - The development of renal cysts appears to be a common feature of terminal renal failure in patients with diffuse renal parenchymal disease. In the present investigation, the kidneys of 13 patients with terminal renal failure but not receiving dialysis, of 14 patients on maintenance hemodialysis and of 4 patients after renal transplantation (patients' own kidneys) were studied by computed tomography. Cystic lesions in the contracted renal parenchyma could be demonstrated by computed tomography in 7/13 non-dialyzed patients with terminal renal failure, in 11/14 patients on maintenance hemodialysis as well as in 3/4 transplanted patients (patients' own kidneys). Both solitary cysts (10/21 patients) and multiple cysts (11/21 patients) were observed. The size varied from 0.5 cm (barely detectable) to 3 cm in diameter. Such cysts could also be demonstrated at autopsy. Possible clinical complications include spontaneous retroperitoneal hemorrhage, macrohematuria, matrix stone formation and formation of benign or malignant papilloma. The present study shows that computed tomography allows the detection of acquired renal cysts in uremic patients in vivo. The cysts appear prior to dialysis, seem to increase in frequency during dialysis and do not disappear after transplantation. The lesions can be distinguished from multicystic or polycystic disease. PMID- 7471533 TI - Immunothrombocytopenia and IgA nephritis. AB - A 32 year old female patient demonstrates the rare combination of a typical IgA nephritis and familial immunothrombocytopenia. Although this association may be purely fortuitous, our observation adds a new facet to a spectrum of reports on (familial) "thromborenal syndromes". The pathogenesis of the IgA nephritis has not yet been clarified. However, since IgA deposits are found relatively frequently in the mesangium in a number of diverse underlying diseases, there might be common etiological factors. In view of this possibility more attention should be directed to the investigation of IgA associated diseases. PMID- 7471534 TI - Dense deposit disease with rapidly progressive renal failure in a narcotic addict. AB - A girl aged 17 developed a nephrotic syndrome with renal insufficiency after narcotic abuse. Renal biopsy showed a diffuse glomerulonephritis with crescents and dense deposits within the glomerular basement membrane; glomerular C3 deposits were present without immunoglobulin. The serum complement profile was typical of activation via the alternative pathway, and tests for C3 nephritic factor were strongly positive. Terminal renal failure occurred within 6 months and required chronic hemodialysis. It is likely that the narcotics used or their contaminants were responsible for the renal damage, presumably by activating the complement system via the alternative pathway. PMID- 7471535 TI - Myocardial metabolism and anginal threshold in coronary disease after verapamil administration. PMID- 7471536 TI - A comparative study of calcium-ion antagonists in patients with variant angina. PMID- 7471537 TI - Clinical pharmacokinetics of calcium ion antagonists. PMID- 7471538 TI - Experimental infarction studies. PMID- 7471539 TI - Assessment of cardioprotective agents. PMID- 7471540 TI - Acute hypertensive crises. PMID- 7471541 TI - Calcium antagonists: mechanisms of action in arrhythmias. PMID- 7471542 TI - Left ventricular function in patients with coronary artery disease after verapamil administration. PMID- 7471543 TI - Effects of intravenously administered verapamil on hemodynamic variables and ventricular function in acute myocardial infarction. PMID- 7471544 TI - Cardioprotective effects of calcium ion antagonists in myocardial ischemia. PMID- 7471545 TI - Symposium. Surgical techniques of the spine. PMID- 7471546 TI - Urogenital complications of anterior approaches to the lumbar spine. AB - The superior hypogastric plexus of the sympathetic nervous system is the only major innervation of the urogenital system which is normally at risk in anterior exposures of the lower lumbar spine. When this is injured, one can expect to see disturbances of urogenital function with retrograde ejaculation or sterility in males. Failure of penile erection is not anticipated unless the patient has, in addition, advanced peripheral vascular disease. The superior hypogastric plexus may be spared by careful dissection about the iliac arteries and lumbosacral junction or by approaching the spine laterally through a retroperitoneal exposure. PMID- 7471547 TI - Unstable hangman's fractures. AB - One hundred and seventy consecutive multiple trauma victims were studied postmortem by radiography and direct inspection at autopsy for the presence of cervical spine injuries. Of the total 38 fractures and/or dislocations, eight had bilateral pedicle fractures of the axis. Only atlanto-occipital dislocations were more frequently encountered. Dissection of six of the eight spines with hangman's fractures showed variable osseous and ligamentous injuries. In only two of the eight cases were significant ligamentous structures sufficiently intact to prevent gross displacement on flexion and extension. In three of the specimens complete laceration of the upper cervical spinal cord was found. Unstable hangman's fractures are a common fatal cervical spine injury in multiple trauma victims. PMID- 7471548 TI - Acute spinal cord injury: current concepts. AB - Optimal treatment of acute spinal cord damage requires an effective emergency medical service at the scene of injury coordinated with a hospital-based multidisciplinary team of physicians and allied health professionals. A detailed protocol outlines the steps required for evaluation, supportive therapy and stabilization. Emphasis is placed on team work with equal attention directed to nervous tissue and spinal column lesions. Laboratory and clinical investigations regarding spinal cord injury are directed toward epidemiology, anti-inflammatory agents, biomechanics, physical therapy and reconstructive surgery. PMID- 7471549 TI - Chemosterilized autolyzed antigen-extracted allogeneic (AAA) bone matrix gelatin for repair of defects from excision of benign bone tumors: a preliminary report. AB - In a series of 33 bone graft operations, antigen-extracted, autolyzed, allogeneic (AAA) bone matrix gelatin was substituted for autologous bone. The period of follow-up was 2.0 to 3.5 years. AAA bone gelatin was resorbed more rapidly than whole bone. AAA bone gelatin was replaced by new bone in the same intervals of time as observed with autologous bone. In treatment of bone tumors with AA bone gelatin, the results of the operation depend upon the nature of the pathologic processes in the host bed. In normal host bed, tight contact between implant and recipient bone is essential for success. The overall results of a preliminary study of 33 cases of 91% successful. PMID- 7471550 TI - Juvenile kyphosis (Scheuermann's disease): radiography, histology, and pathogenesis. AB - In juvenile kyphosis (JK), the roentgenographic defects correspond to the sites of large translucent areas of the collagen-free tissue in the cartilaginous end plates of vertebral bodies. The extent of the roentgenographic lesions is proportional to the number and size of these translucent areas. The gradual transition of radiologic signs between the fully developed JK and the normal spine is paralleled by the decreasing intensity of these histologic change. This relationship can be traced from individual to individual, and within single vertebral columns. The growth zone is often narrow or even missing. The end plates are narrow and segmentally indented towards the vertebral body. These findings are manifestations of the pathogenetic process. Prolapses of disk tissue through gaps in the end plates are probably secondary to the loss of mechanical strength in these defective areas. PMID- 7471551 TI - Clinical experience with the technetium-99 scan in children. AB - A study of 86 bone scans in 79 patients was conducted to determine ways to enhance the usefulness of this test and learn what factors influence its results. While the overall value of the bone scan as we initially used it was not what we had hoped, it could be improved substantially by utilizing all initial clinical information when ordering as well as interpreting the scan. Its main value in suspected sepsis is to localize affected areas in and around the pelvis and spine. The scan is often unnecessary in the diagnosis of long-bone osteomyelitis and is seldom indicated when diagnosing cellulitis or synovitis. In occult trauma it is valuable for further investigation of specific areas with possible nondisplaced fractures. Needle aspiration of the bone or joint does not seem to alter the scan. We conclude that the bone scan is chiefly valuable in locating focal areas of skeletal pathophysiology. PMID- 7471552 TI - A rational approach to burst fractures of the atlas. AB - The stability of a burst fracture of the atlas (Jefferson's fracture) should be determined in order to decide upon the appropriate treatment. Unstable fractures include those with a rupture of the transverse ligament, or those in which union has been incomplete despite treatment. An occiput-to-C2 or in atlantoaxial arthrodesis may be indicated. PMID- 7471553 TI - Elbow dislocation during overhead skeletal traction therapy: a case report. AB - A dislocation of the elbow occurred in a 7-year-old boy during overhead skeletal traction treatment for a supracondylar fracture of the humerus. Closed reduction of the dislocation was successful. A review of the literature failed to reveal a report of a similar incident. PMID- 7471554 TI - Irreducible dislocation of a distal interphalangeal joint: case report and review of literature. AB - A 12-year-old boy incurred an irreducible dislocation of a distal interphalangeal joint of the hand secondary to volar plate interposition. To our knowledge, only five cases of this traumatic lesion have been previously described. PMID- 7471555 TI - Containment of the hip: a theoretical comparison of osteotomies. AB - Using a computerized model derived from anatomic specimens, coverage of the femoral head by the acetabulum during the gain cycle was studied and compared for normal hips, varus-derotation osteotomy, and innominate osteotomy (Salter). Theoretical estimates of joint reaction force were also studied during the gain cycle for the three conditions. The entire proximal femoral epiphysis was never covered by the acetabulum with any of these procedures. Innominate osteotomy gave better anterior coverage (with resultant poorer posterior coverage) than the femoral procedure. Varus-derotation osteotomy increased medial shear force at the epiphyseal plate to twice body weight, but in all cases the major component of force was normal to the epiphyseal plate (compression) rather than parallel to it (shear). PMID- 7471556 TI - Infected late false aneurysm of the deep femoral artery. AB - A false aneurysm may mimic an abscess, but concomitant existence of the two lesions seems not to have been previously described. In a 32-year-old man, a false aneurysm of the deep femoral artery was found to be in communication with an abscess cavity in the posterolateral thigh. The false aneurysm was found in the proximity of the protruding tip of a Steinmann pin which had eroded the arterial wall. The abscess was a consequence of posttraumatic and postoperative osteomyelitis which had been previously debrided, bone grafted, and immobilize with an external fixator attached to four Steinmann pins. The condition was successfully treated by surgical drainage of te abscess cavity, excision of the false aneurysm, and ligation of the deep femoral artery. PMID- 7471557 TI - Complications of posterolateral dislocation of the knee: case report and literature review. AB - A 49-year-old man incurred a posterolateral dislocation of the knee, a rare type of knee injury. The injury was associated with a significant skin slough as a result of internal pressure from the displaced medial femoral condyle. The dislocation also was associated with a rupture of the quadriceps tendon. The mechanism of injury was a forcible abduction of the flexed knee, coupled with medial rotation of the tibia upon the femur. This type of knee dislocation cannot be reduced closed and therefore immediate open reduction with primary ligamentous repair is the treatment of choice. PMID- 7471558 TI - Acquired valgus instability after knee replacement. AB - Four cases of acquired valgus instability occurred after knee resurfacing operations; all four were initially stable. Three cases originally had a varus deformity which required extensive releases for its correction. All of these patients fell, disrupting the medial joint capsule and medial collateral ligament. This complication in the two early cases could have been prevented by adequate bracing until full recovery of the quadriceps had occurred. In one case, attribution due to projection medially of the tibial component may have played a role. In the fourth case, the tibial component was inserted with the tibia in external rotation, thus advancing the tibial attachment of the medial collateral ligament, which was disrupted during manipulation to gain flexion. The simplest treatment for this complication is probably to brace the knee until the joint reaction settles and then revise to a fully constrained prosthesis. PMID- 7471559 TI - Positioning techniques in spinal surgery. AB - By keeping in mind the principle of stability, ease of exposure and physiologic restrains on the patient, the optimum safe positioning technique for successful spinal surgery can be selected. We recommend skeletal or head-halter traction for anterior exposure of the cervical spine. Gardner's three-point skeletal fixation or halothoracic immobilization are the methods of choice for posterior cervical spine exposure. Either rolled towels or the Relton-Hall frame are preferred for posterior thoracolumbar surgery. Specific attention to protection of the eyes, bony prominences and peripheral nerves is essential. PMID- 7471560 TI - Development of maduromycosis (Madurella mycetomi) after nailing of a closed tibial fracture: a case report. AB - Maduromycosis is a rare fungal infection in the northern United States, most frequently caused by Allescheria boydii, usually in debilitated or immunosuppressed patients. An 18-year-old Mexican-American, in excellent health, developed multiple draining sinuses at the sites of insertion of Ender nails for a closed tibial fracture. Madurella mycetomi was cultured from black granules in a biopsy specimen. The source of fungal infection remained unknown, although it probably did not occur during the surgical procedure. Since no specific antibiotic treatment is available, it is possible that with progression of the infection, amputation may be necessary in the future. PMID- 7471561 TI - Early Dwyer operation in talipes equinovarus. AB - For clubfeet resistant to conservative treatment, an early soft-tissue release and reduction of talonavicular dislocation are recommended. To prevent the deforming influences towards varus of the often small inverted heel and make it reach the ground properly, a Dwyer osteotomy of the calcaneum is indicated. In a series of 22 patients this operation was performed on 29 clubfeet. The results at follow-up were satisfactory in all of the patients operated on before the age of 1 year, the youngest being 3 months. Young patients with a varus and small inverted heel respond very well. PMID- 7471562 TI - Giant-cell tumor of the second cervical vertebra treated by cryosurgery and irradiation. AB - A 17-year-old female presented with a large, destructive, giant-cell tumor of the second cervical vertebra. The lesion was obviously life-threatening. Owing to its proximity to the spinal cord and the danger of a pathologic fracture with cord transection and because of the effectiveness of cryosurgical methods in treating giant cell tumors of long bones, we decided to attempt local freezing of the lesion using a cryoprobe via a transoral approach. To further ensure tumor destruction, we followed cryosurgery with a course of radiotherapy (4600 rads). At this writing, 30 months later, healing has been excellent with no signs of recurrence, pain, tenderness or neurologic problems. To the best of our knowledge, this represents the first time a vertebral body bone tumor has been treated by cryosurgery. PMID- 7471563 TI - Distraction epiphysiolysis as a method of limb lengthening. I. Experimental study. AB - The tibiae of 41 growing sheep were lengthened by distraction epiphysiolysis. The distraction apparatus was applied for varying periods of time to the proximal epiphysis of a tibia; the contralateral tibia was used as the control. Daily application of the distraction force initially induced a fracture within the growing cartilage, and then progressively increased the distance between epiphysis and metaphysis. In the newly created space, new bone tissue gradually formed, which in time developed into a neometaphysis with a new cortex. This neometaphysis gradually acquired radiographic characteristics resembling those of control tibia, and they were capable of supporting normal body weight. Elongation of the bone ranged between 30 and 100 mm. Except in one incident of sepsis, no significant changes were observed in the structures adjoining the elongation area, nor in the soft tissue. In general, bone growth was not affected, although premature fusion of the epiphysis occurred in a few cases. PMID- 7471564 TI - Hypotensive anesthesia and autotransfusion in spinal surgery. AB - Sixty-eight (68) spinal operations performed under normotensive anesthesia were compared with 77 procedures performed under controlled hypotensive anesthesia. Procedures reviewed included Harrington instrumentation and fusion, dorsal arch resection, laminectomies and other spinal fusions. Intraoperative and postoperative blood loss and blood replacement were found to be reduced by approximately 50% in the group undergoing controlled hypotensive anesthesia for each procedure reviewed. This reduction in blood loss was achieved with only a moderate reduction in blood pressure (20 mm Hg systolic). Reduction in systolic blood pressure of greater than 20 mm was not associated with greater reduction in blood loss. Deliberate hypotension was coupled with hemodilution, lowering the hematocrit to 28% to 30% in order to increase cardiac output, increase tissue perfusion and decrease venous stasis. Autotransfusion has further reduced the need for homologous transfusion such that the combined techniques of hypotension and autotransfusion can eliminate the need for homologous blood transfusion in all but the most unusual cases. PMID- 7471565 TI - Distraction epiphysiolysis as a method of limb lengthening. III. Clinical applications. AB - In 16 patients still in growth stage who were suffering from a shortened limb from 5 to 12 cm attributable to a varied etiology, the tibia was lengthened by distraction, hence, achieving separation of the epiphysis (distraction epiphysiolysis). Lengthening of the tibia from 5 to 10 cm was attained with remarkably minor and infrequent complications. The fact that distraction epiphysiolysis is a blood less procedure, that it allows the formation of a bone segment which is qualitatively and quantitatively suitable to its functional needs, and that all of this is achieved in a reasonably short period, justifies the conclusion that this method is a valuable addition to the accepted techniques for limb lengthening. PMID- 7471567 TI - Chevron osteotomy. PMID- 7471568 TI - Abstracts: Transactions of the thirty-second annual meeting of the Association of Bone and Joint Surgeons, Durango, Colorado, June 1980. PMID- 7471566 TI - Experimental femoral fracture immobilized by rigid and flexible rods (a rabbit model). AB - Experimental fractures in rabbits demonstrated that flexible intramedullary rod fixation of the midshaft long bone fracture enhances the fracture healing and may promote early mobility. When compared to rigid fixation, both excess flexibility and excess rigidity inhibited optimal fracture healing. The occurrence of angulation deformities with plastic rods also demonstrate the disadvantage of a very flexible rod. PMID- 7471569 TI - An inexpensive functional fracture table. PMID- 7471570 TI - Pin benders. PMID- 7471571 TI - Anterior drawer sign. AB - The traditional method for testing for anterior stability of the knee has often lead to inaccurate assessment of ligament stability because of the jerky nature of the test and the small amount of force applied. A method of testing the anterior drawer of a knee by flexing the hip and knee to 90 degrees and applying a steady upward force using the body weight as counterpull provides a more reliable test for assessing ligament stability. This method can be used to overcome hamstring spasm and causes less pain in the acutely injured knee. PMID- 7471573 TI - Replacement of the displaced guidewire for sliding compression nails in proximal femoral fractures. PMID- 7471572 TI - Beck wire for passing sutures through bone. PMID- 7471574 TI - A useful pair of retractors for small bone surgery. PMID- 7471575 TI - Obtaining iliac graft for the Russe procedure. PMID- 7471576 TI - Transfixation pin removal: a new twist. PMID- 7471577 TI - The plaster slipper cast. AB - Since cast correction of congenital metatarsus adductus requires purchase of the cast on the foot only, a plastic slipper cast can accomplish this task. The correction of deformity and retainability equals or exceeds that of the short-leg cast. Ease of application and removal make this cast quite attractive in the small child. The use of this cast can be extrapolated to many conditions. PMID- 7471578 TI - A convenient distal plug for the femoral cavity in total hip arthroplasty. AB - A new technique for plugging the femoral cavity prior to inserting methylmethacrylate during total hip arthroplasty is described. The plastic "Mexican hat" cement restrictor previously used in the acetabulum is trimmed and used conveniently as the femoral plug. This technique has been used satisfactorily and without complication in more than 50 cases. PMID- 7471579 TI - Arthroscopic surgery: simple devices to facilitate multiple entries and manipulations of the knee. AB - A system has been especially designed and fabricated for use with an arthroscope and other instruments so that an open passage is maintained through the skin, retinaculum, and joint capsule to allow the insertion and withdrawal of instruments at will. This system is designed to minimize loss of irrigating fluid from the joint during use, including instrument exchanges. A companion stabilization device has been constructed to allow the unassisted application of valgus or varus stress to the knee while leaving the surgeon's hands free. PMID- 7471580 TI - Technique of windowing the femoral shaft for removal of bone cement. PMID- 7471581 TI - Knee joint puncture after patellectomy. PMID- 7471582 TI - The pediatric spica cast: a sex-related modification. AB - For the child's comfort and to facilitate nursing care, placement of the supporting strut on a hip spica is posterior for the male and anterior for the female. PMID- 7471583 TI - Cervical orthoses: a guide to their selection and use. AB - A large variety of cervical orthoses is available, but these may be divided into four basic groups. Although the orthoses in each group provide similar controls, each appliance has certain discrete advantages and limitations. The effectiveness of seven different cervical appliances in restricting motion in flexion extension, lateral bending and rotation is presented. This information may be used to rationally select an orthosis to control specific clinical problems. A guide is formulated for selecting the orthoses for the control of various cervical injuries and postoperative problems. PMID- 7471584 TI - The stone table staple modification for obtaining firm bone fixation. PMID- 7471586 TI - Intraoperative straight-leg raising during laminectomy and disk excision for sciatica. AB - During lumbar disk excision for sciatica, doubts may occasionally arise as to the level of the lesion or the degree of involvement of the particular nerve root being observed. Following removal of the obstructing agent, can we be certain that liberation of the nerve root is complete? These anxieties can be dispelled by performing a straight-leg raising test bilaterally during the operation both before and after removal of the obstructing agent. The intraoperative straight leg raising tests enable observations to be made as to the degree of nerve tension on the involved side as compared with tension on the uninvolved side; at the end of the operation, the tension on the involved side should be no more than the tension on the uninvolved side. When the straight-leg raising test is not being performed, it should be possible for the nerve root to be displaced to the midline of the spinal canal with ease. PMID- 7471585 TI - Intraoperative use of nylon bands in fracture fixation. AB - Nylon tie bands are introduced as an intraoperative aid in the application of internal fixation. Their use for holding metal plates on reduced fractures is advantageous over metal clamps in that they do not impede access to the fracture site, are easily applied, keep intraoperative injury to soft tissue at a minimum, and reduce manipulatory efforts at the fracture site, decreasing operating time. Scuffing of metal plates is eliminated. A technique for their use is described. Our experience with the nylon tie bands indicates that they are an effective adjunct, as opposed to conventional metal clamps, in open reduction and internal fixation of fractures in our institution. PMID- 7471587 TI - A reducer for closed femoral rodding. PMID- 7471589 TI - The use of intraoperative fluoroscopy and spinal cord monitoring for transoral microsurgical odontoid resection. AB - The transoral approach to the upper cervical spine has not gained wide acceptance. A reliable intraoperative method to determine the necessary extent of dissection and degree of decompression, as well as to evaluate spinal cord function, should improve the safety and applicability of the transoral approach. Four cases of anterior cervical cord compression were successfully treated by odontoid resection employing a microsurgical transoral approach. Intraoperative fluoroscopy monitored the depth of dissection and adequacy of decompression. Somatosensory cortical evoked potentials (SSEP) allowed assessment of spinal cord function that could be correlated with surgical decompression. In addition, follow-up examination revealed that SSEP was an intraoperative prognosticator of subsequent neurologic improvement. PMID- 7471588 TI - Indications for the use of the halo immobilization. AB - Since its early development, the indications and utilization of the halo cast have increased to the point where it has become a versatile adjunct in the armamentarium for treatment of spinal injuries, diseases and deformities. Its prime indication continues to be that of traction in the cervical spine and in spinal deformities. The ease of application and low morbidity in its use for traction or immobilization render the halo splint an incomparable aid in the treatment of spinal deformities and injuries. PMID- 7471590 TI - Surgical techniques of anterior decompression and fusion for spinal cord injuries. AB - Many patients who have static or only slowly improving neurologic deficits and significant compression of the spinal cord and nerve roots can benefit from anterior decompression. The improvement ranges from partial root recovery to very dramatic improvement in upper as well as lower extremities in the patient with quadriparesis. Intrinsic damage or contusion of the spinal cord cannot be reversed by decompression. Patients with motor sparing preoperatively attain a better functional improvement than those patients who have only slight distal sensory function initially. The same can be said of patients with spinal cord injuries treated with surgery, but we believe the ultimate degree of functional recovery of incomplete cord injuries is greater following anterior than posterior decompression when the operation is indicated. An early accurate diagnosis must be made concerning whether a patient has a complete or an incomplete spinal cord injury. The mechanical compressive lesion must be well documented by myelography, laminography, or CAT scan. The patient should not be neurologically harmed by a posterior laminectomy approach to anterior pathology which additionally removes all posterior stability. An anterior compressive block is best removed through an anterior approach. PMID- 7471591 TI - Spondylolisthesis: treatment by laminectomy and posterior interbody fusion. AB - Spondylolisthesis is a hereditary anomaly of the spine, often associated with intractable pain in the back and lower extremities. The diagnosis is commonplace. Approximately 5% to 7% of the population in the United States has the anomaly. This percentage varies with country and race. In patients who seek medical aid for symptoms of low back pain, 10% will have a pars defect. Laminectomy of the entire separate neural arch and a posterior interbody fusion is a superior operation for spondylolisthesis. It is a difficult operation, requiring a high degree of technical skill, but when properly performed, it is possible to attain a more rapid recovery with a lower morbidity and higher percentage of permanent cures. PMID- 7471592 TI - Neural arch resection for lumbar spinal stenosis. AB - In a series of 50 patients with lumbar stenosis who have undergone the procedure of neural arch resection, the patients were predominantly in the fifth and sixth decades of life. Females exceeded males 3:2. Long histories and previous surgical treatment were characteristic, with unilateral symptoms predominating. However, physical examination demonstrated the reverse, i.e., bilateral signs and multisegmental disease. Lumbosacral radiographs suggested the stenotic condition and myelography was corroborative. The definitive test was axial tomography. Neural arch resection removes, bilaterally, the spinous processes, laminae, articular processes and the pedicles, if the latter are hypertrophied or narrowly placed. Resection must be accomplished at all levels of disease for optimum results. Fusion has not been required. Half of the group of 50 totally disabled patients have been returned to full function and comfort, one-fourth to perform acts of daily living with some pain, and one-fourth remain disabled. Complications include infections in six and one case of postoperative spondylolisthesis. Neural arch resection is a procedure for the management of lumbar stenosis and its elemental categories. The results indicate at least a 50% expectation of return to function in comfort, provided that the principles of surgery as to extent and removal of disease at all levels are followed, and that the patients participate in a back rehabilitation and conditioning program. PMID- 7471593 TI - Spinal cord decompression in spinal cord injury. AB - There is substantial doubt as to the value of "decompressive" surgery in the management of spinal cord injury. A few relative indications exist for such surgery, but they are only relative. There are a number of absolute contraindications to the procedure, and it should be undertaken only under the most unusual and highly selected circumstances. It would seem that stabilization of the spine may be more important. It behooves us to look more carefully at our concepts of "stable" versus "unstable" fractures. PMID- 7471594 TI - Assessment of evaluation of hospitalized pediatric patients with genetic disorders. AB - A survey of the status of pediatric and neonatal care in a university general hospital was conducted for a 12-month period in order to define existing needs and plan for improved inpatient genetic management. The records of 218 patients with one of 22 specified disorders were reviewed to evaluate the nature of documented genetic considerations and formulated dispositions during hospitalization. The 22 clinical entities were classified as chromosomal, single gene, polygenic, or possibly genetic. In 15.1 per cent of cases the need for genetic counseling was recognized and recorded on the patient's chart. Sixteen of the 218 records (7.3%) met the minimum criteria for a "genetic disposition." Suggestions for improving the hospital clinical management of patients with genetic disorders are presented. PMID- 7471595 TI - Respiratory syncytial virus infection: ten-year follow-up. AB - Studies within the last 20 years have shown that the incidence of asthma is increased in children who have been hospitalized with bronchiolitis due to respiratory syncytial virus. To evaluate such respiratory consequences in an ambulatory population, a follow-up study was done of 37 children who had respiratory syncytial virus infection documented at a community pediatric clinic in 1968 when they were younger than 4 years. This study, done ten years later by chart review and parental interview, showed that asthma had subsequently been diagnosed in three (8%). Thus, an increased risk for the development of asthma after respiratory syncytial virus infection was not found. Comparison of these data with those from previous reviews questions a causative role for respiratory syncytial virus in childhood asthma. PMID- 7471596 TI - Fever in Haemophilus influenzae type B meningitis. AB - Ninety-nine cases of Haemophilus influenzae type B (HIB) meningitis were analyzed to assess the causes of secondary and persistent fever and outcome of treatment. Secondary fever developed in 47 patients, 11 had persistent fever and 4 patients died. Secondary fevers were associated with intercurrent illness in 30 per cent, drug fever in 15 per cent, and neurologic complications in 13 per cent. Serious neurologic or other complications were no more common in patients with than in those without secondary fever. However, patients with persistent fever demonstrated a higher rate of neurologic complications. Neither persistent nor secondary fever are markers of bacteriologic relapse or inadequate therapy. PMID- 7471597 TI - Pseudomembranous colitis following obstruction in a neonate. PMID- 7471598 TI - Campomelic syndrome in phenotypic females with 46,XY chromosomes: evidence of genetic heterogeneity. PMID- 7471599 TI - Sudden death due to colloid cyst of the third ventricle. PMID- 7471600 TI - Effects on exercise tachycardia during forty-eight hours of a series of doses of atenolol, sotalol, and metoprolol. AB - Beta adrenoceptor blockers differ mainly in their plasma elimination half-lives (t 1/2 s). It has been assumed that drugs with longer t 1/2 will have a longer duration of effect on exercise tachycardia. Several factors may influence the duration of action of beta blockers; we have investigated the contribution of plasma elimination t 1/2 and dose by comparing the effects on an exercise tachycardia in healthy subjects of placebo, 25, 50, 100, and 200 mg of atenolol and of sotalol, and 50, 100, 200, and 400 mg metoprolol. Subjects exercised before and at 2, 3, 6, 8, 24, 33, and 48 hr after oral doses of each drug. Plasma samples for measurement of drug concentration were drawn before each exercise period. Twenty-four hours after 50, 100, and 200 mg atenolol and 50, 100, 200, and 400 mg sotalol there were reductions in an exercise tachycardia; at this time reductions were greater after the larger doses. The plasma elimination t 1/2s of atenolol were between 7.2 +/0 1.0 hr. Although 50, 100, and 200 mg metoprolol induced the same reductions in an exercise tachycardia 2 hr after drug as 25, 50, and 100 mg atenolol and 50, 100, and 200 mg sotalol, these doses were without effect at 24 hr. Metoprolol 400 mg reduced exercise tachycardia at 24 hr but the effect was less than that of the three largest doses of atenolol and sotalol. The plasma elimination t 1/2 for metoprolol was between 3.6 +/- 0.6 and 5.0 +/- 1.8 hr. These results show that duration of cardiac beta blocking of cardiac beta blocking activity of atenolol, sotalol, and metoprolol is determined by the elimination t 1/2 and dose. PMID- 7471601 TI - Effect of large oral doses of ascorbic acid on uric acid excretion by normal subjects. AB - The effects of large and oral doses of ascorbic acid on renal clearance and excretion of uric acid were studied in nongouty subjects because ascorbic acid has been reported to increase renal uric acid clearance. Our results indicate that 4 or 12 gm ascorbic acid taken in divided doses had no effect on serum uric acid concentration or uric acid excretion and clearance by the kidney. Reasons for these results, which differ from previous reports, are discussed. We quantitated the magnitude of the interference of ascorbic acid in the measurement of uric acid by the nonspecific methods frequently used, since falsely elevated urine uric acid could lead to misinterpretation of screening tests. PMID- 7471602 TI - Disopyramide hemodialysis and kinetics in patients requiring long-term hemodialysis. AB - Disopyramide hemodialysis and kinetics after 200 mg orally in six patients receiving long-term hemodialysis were examined. Mean volume of distribution (area) was 66.5 +/- 13 l. Mean times of the peak serum concentration and mean peak serum concentration were 2.3 +/- 0.9 hr and 3.1 +/- 0.9 microgram/ml. Mean absorption half-life (t 1/2) was 21.6 +/- 12.5 min. Mean disopyramide elimination t 1/2 during dialysis was 16.8 +/- 11.9 hr, not significantly different from mean elimination t 1/2 without dialysis of 16.1 +/- 5.2 hr. End-dialysis bath concentrations of disopyramide showed that not more than 2.4% of the dose was dialyzed during a 2-hr dialysis period. Our data indicate that at therapeutic concentrations disopyramide was not appreciably dialyzed. PMID- 7471603 TI - Dynamic interaction between disulfiram and separated enantiomorphs of racemic warfarin. AB - To evaluate the interaction of disulfiram with the separated enantiomorphs of racemic warfarin, seven normal subjects received single doses of R-warfarin (1.5 mg/kg body weight) and S-warfarin (0.75 mg/kg body weight) with and without a daily dose of disulfiram 250 mg, beginning 3 days before the warfarin dose and continuing for the duration of the hypoprothrombinemia. Disulfiram augmented the S-warfarin hypoprothrombinemia (p less than 0.001) but not that of R-warfarin (p less than 0.10). Disulfiram did not alter plasma concentrations of either R warfarin (p greater than 0.10) or S-warfarin (p greater than 0.40). Disulfiram augments the hypoprothrombinemia of racemic warfarin stereoselectively by interacting primarily with S-warfarin. As disulfiram did not change the plasma concentrations of either enantiomorph, it may augment the anticoagulant effect of racemic warfarin by directly affecting the hepatic mechanism responsible for the hypoprothrombinemia. PMID- 7471604 TI - Effect of dose on acetylator phenotype distribution of hydralazine. AB - The effect of dose on acetylator phenotype distribution of hydralazine has been determined, The acetylated metabolites methyltriazolophthalazine (MTP) and 3 hydroxymethyltriazolophthalazine (3-OHMTP) and acid-labile hydralazine (HP) were determined in the 0- to 24-hr urine of patients receiving various doses. The difference between the mean value for the ration 3-OHMTP:HP in the rapid and slow acetylators varied with dose, the greatest difference being after a 200 mg (100 mg twice daily) dose. The distribution of the ratio became less clearly bimodal at lower doses, with overlap between phenotypes occurring at doses of 100 mg (50 mg twice daily) or less. The most effective dose for discriminating between acetylator phenotypes was found to be 200 mg (100 mg twice daily). PMID- 7471605 TI - Enhancement of uricosuric properties of indacrinone by manipulation of the enantiomer ratio. AB - Racemic indacrinone is a high-ceiling, relatively long-acting diuretic. Both enantiomers have uricosuric activity, but the diuretic activity resides predominantly in the (-) enantiomer. Usual therapeutic doses of racemic indacrinone have only transient uricosuric activity, so that, as with other diuretics, hyperuricemia occurs. Sixty-five healthy men participated in a multicenter, double-blind, randomized, balanced, incomplete-block study comparing the effects on plasma urate and urate clearance of indacrinone (-) enantiomer 10 mg given concomitantly with 0, 10, 20, 40, and 80 mg (+) enantiomer (10/0, 10/10, 10/20, 10/40, 10/80), as single daily doses for 7 days. Hydrochlorothiazide (HCTZ) 50 mg daily and ticrynafen (T) 250 mg daily were controls. Each subject received two of the seven treatments, so that there were 18 subjects per treatment. On days 7 to 8, morning (mean of 0-hr values on days 7 and 8), HCTZ, 10/0, 10/10, and 10/20 elevated plasma urate by 8% to 16%. 10/40 was approximately isouricemic, and 10/80 and T lowered plasma urate by 13% and 41%. There were corresponding changes in urate clearance. PMID- 7471606 TI - Probenecid-clofibrate interaction. AB - Clofibric acid disposition was studied in four healthy men after 1 wk of clofibrate ingestion (500 mg orally every 12 hr) with and without probenecid (500 mg orally every 6 hr). Mean (+/- SD) free clofibric acid plasma concentration in the four subjects over a dosage interval at steady state was 2.5 +/- 0.03 mg/1 before and 9.05 +/- 1.09 mg/1 after the probenecid. Probenecid reached an average plasma concentration of 71.3 mg/1. No clofibric acid glucuronide was detected in plasma during either treatment. The fractions of the dose recovered in urine as clofibric acid, clofibric acid glucuronide, and clofibric acid liberated after acid hydrolysis were not altered by probenecid. These data suggest that probenecid causes a reduction in renal and metabolic clearance of clofibric acid, probably as a result of inhibition of the conjugation of clofibric acid with glucuronide. PMID- 7471607 TI - Clearance and recovery calculations in hemodialysis: application to plasma, red blood cells, and dialysate measurements for cyclophosphamide. AB - The hemodialyzability of cyclophosphamide was investigated in four patients on long-term hemodialysis. Cyclophosphamide 100 mg was given intravenously over 10 min before hemodialysis. Blood and dialysate samples were collected periodically during the 4 hr dialysis and measured by gas-liquid chromatography (GLC) for cyclophosphamide. Dialysis clearance calculated by arterial-venous difference and actual drug recovery in dialysate averaged 104 ml/min, which is in the range of the metabolic clearance of 95 ml/min for the drug. The extraction efficiency of the hollow-fiber dialyzers averaged 40% for plasma and red blood cell (RBC) samples. A mean of 37% of the administered dose of cyclophosphamide was removed during hemodialysis. The half-life (t 1/2) of the beta phase was 3.3 hr in our patients during hemodialysis, a 49% reduction of the 6.5 hr to t 1/2 reported in uremic patients. Because of the reduction in elimination t 1/2, larger dialysis clearance than metabolic clearance, high extraction efficiency, and significant drug removal during dialysis, we conclude that cyclophosphamide is dialyzable. PMID- 7471608 TI - Disposition of anagrelide, an inhibitor of platelet aggregation. AB - A single dose of 14C-anagrelide (6,7-dichloro-1,5-dihydroimidazo [2,1-b] quinazoline-2(3H)-one monohydrochloride) equivalent to 1 mg free base and containing 100 muCi radioactivity, was taken by five healthy, fasting men. Blood, plasma, urine, and feces were analyzed for total radioactivity. Plasma and urine concentrations of anagrelide were determined, and the urinary metabolite profile was established by high-performance liquid chromatography (HPLC). The drug was rapidly absorbed with peak plasma levels of radioactivity equivalent to 50 ng anagrelide per milliliter at about 1 hr. These levels decreased to less than 10% of peak in 24 hr. Plasma levels of anagrelide peaked at 5 ng/ml at about 1 hr, decreased rapidly during the first 6 to 8 hr, and then declined more slowly, with an estimated terminal elimination half-life of about 3 days. No significant quantities of radioactivity were associated with the cellular elements of blood. Anagrelide was extensively metabolized before elimination in urine. Means of 68% and 72% of the dose were excreted in urine as metabolites in 24 and 144 hr, and 10% of the dose recovered in the feces. Several urinary metabolites were detected by HPLC. PMID- 7471609 TI - Heparin kinetics: variables related to disposition and dosage. AB - A method to determine heparin kinetics and dosage requirements was examined in 20 patients with active thromboembolic disease. Pretreatment heparin sensitivities were determined to establish the relationship between heparin concentration and activated partial thromboplastin times (APTTs). After an initial bolus dose, serial APTTs were measured, heparin concentrations were estimated, and kinetic determinations followed. Heparin elimination rate, distribution volume, and clearance were used to calculate dosage requirements. There was a 500% range in pretreatment heparin sensitivities. Smokers had more rapid heparin elimination rates and t 1/2s than nonsmokers did. Men had more rapid drug clearances than women did. Body weight was related to heparin dosage requirements. Patients treated early after onset of symptoms required higher doses than patients in whom treatment was delayed. A multiple regression model was developed for heparin dosage requirements from body weight, sex, delay between onset of symptoms and treatment, and smoking. This statistical model explained 78% of the variance in heparin requirements. PMID- 7471610 TI - Amrinone metabolism. AB - High-performance liquid chromatographic methods for the analysis of amrinone in plasma and for both amrinone and its N-acetyl metabolite in urine were developed and applied to measure specimens obtained from a number of healthy men who had received intravenous or oral amrinone. The intravenous doses ranged from 0.8 to 2.2 mg/kg. Terminal elimination of amrinone from the bloodstream followed apparent first-order kinetics. Half-life, after the drug had distributed to the tissues, was estimated by a log-linear least-squares regression; mean half-life was 2.6 +/- 1.4 hr. During the first 24 hr after medication, unchanged amrinone excreted in the urine of these subjects represented 10% to 40% of the dose. N Acetyl metabolite in the urine represented less than 2% of the dose. In the oral study, doses ranged from 25 to 250 mg (0.31 to 3.5 mg/kg) and the maximum plasma concentration attained was proportional to the dose. The first order terminal elimination half-life was possibly dose-related. In only one subject were there unequivocal amounts of the N-acetyl metabolite in the plasma. PMID- 7471611 TI - Primidone kinetics: effects of concurrent drugs and duration of therapy. AB - Primidone (PRM) kinetics was examined in two groups of adult seizure patients: (1) 10 newly diagnosed in whom only PRM was used, the monotherapy (MT) group, and (2) nine in whom PRM was added to other antiepileptics, the combination therapy (CT) group. Time-concentration data were obtained after an initial dose of 250 mg and during subsequent steady-state periods. PRM elimination was slower (p less than 0.05) after the initial dose in MT patients (half-life (t 1/2) = 15.2 hr, apparent clearance = 35 ml/hr/kg) than in CT patients (t 1/2 = 8.3 hr, clearance = 51 ml/hr/kg). PRM metabolites, phenobarbital and phenylethylmalonamide, appeared much earlier in CT patients. Continued PRM exposure in MT patients was accompanied by an increase in apparent clearance in three of seven patients, but no change in four of seven. In four CT patients in whom other antiepileptics were withdrawn there was a decrease in apparent clearance (61.4 to 29.9 ml/hr/kg) no rates in the range of MT patients. PRM kinetics is influenced by concurrent antiepileptic drugs and by duration of PRM therapy. PMID- 7471612 TI - Sotalol kinetics in renal insufficiency. AB - Kinetics of sotalol, a beta adrenoceptor blocker, was studied in 20 patients with varying renal function. In subjects with creatinine clearance (Clcr) greater than or equal to 39 ml/min/m2, sotalol plasma clearance (x +/- SD) was 71 +/- 31 ml/min/m2, elimination half-life (t 1/2) was 8.1 +/- 3.4 hr, and renal clearance was 46 +/- 26 ml/min/m2. In patients with moderate renal impairment (Clcr = 8 to 38 ml/min/m2) elimination t 1/2 rose to 24.2 +/- 7.5 hr, and plasma clearance fell to 24 +/- 7 ml/min/m2. In patients receiving dialysis, elimination t 1/2 rose to 33.9 +/- 27.1 hr. Elimination t 1/2 during hemodialysis was 5.8 +/- 2.1 hr and was associated with a 56.7 +/- 21% reduction in plasma levels. PMID- 7471613 TI - Meperidine binding in maternal and fetal plasma. AB - Meperidine protein binding was measured in nine pairs of maternal and fetal plasma samples obtained at delivery. For the maternal samples, percent bound and binding ratio (bound/free, B/F) were 63.3 +/- 6.18% (SD) and 1.79 +/- 0.45, and for the fetal samples the corresponding values were 51.7 +/- 4.53% and 1.09 +/- 0.21. In each case the binding was higher in the mother than in the fetus (p less than 0.01). Plasma alpha 1-acid glycoprotein (alpha 1-AGP) concentrations were higher (p less than 0.01) in maternal than in fetal samples, and there was a correlation between meperidine B/F and plasma alpha 1-AGP concentration for the maternal and fetal samples (r = 0.752, p less than 0.01). Binding studies with purified alpha 1-AGP showed that this was a cause-effect relationship. The transplacental binding differential was attributable partially to the maternal fetal difference of plasma alpha 1-AGP concentrations. Meperidine was 17.5 +/- 0.35% bound in a 3.5 gm/100 ml solution of human serum albumin; however, there was an inverse correlation (r = -0.798, p less than 0.01) between B/F and plasma albumin concentration for the maternal and fetal samples. A relatively large proportion (75%) of the overall variability in B/F was accounted for by plasma alpha 1-AGP and albumin. Plasma binding of this basic drug was not greatly influenced by the perinatal levels of bilirubin and nonesterified fatty acids. The common clinical observation of greater fetal than maternal plasma total meperidine concentrations at delivery is not the result of more extensive protein binding in fetal than in maternal plasma. PMID- 7471614 TI - Transsynovial distribution of ibuprofen in arthritic patients. AB - Free and total ibuprofen levels in serum and synovial fluid (SF) were determined in one male and 14 female arthritic patients (mean age 56 yr, range 19 to 77) after 400 mg three times daily for 2 days. Free drug was separated by ultrafiltration and assayed by a new highly sensitive and precise gas chromatographic-mass spectrometric technique. Total protein and albumin were assayed by automatic analyzer, and the routine serum biochemical profile obtained. Serum and SF nonesterified fatty acids (NEFAs) were methylated and assayed by gas chromatography. While both total protein and albumin levels were lower (P less than 0.01) in SF, NEFA levels did not differ (p greater than 0.05). Ibuprofen readily partitions into and binds strongly to joint fluid, the total level being about half that in serum. The ratio of total ibuprofen in SF to that in serum (x = 0.41 +/- 0.17) correlated with the albumin concentration ratio (r = 0.886) indicating that albumin binding is an important determinant of ibuprofen transsynovial distribution. A low total ibuprofen SF/serum ratio, apparent for three patients on concomitant aspirin therapy, is consistent with reports of an ibuprofen-salicylate interaction. Free ibuprofen in SF (0.192 +/- 0.086 microgram ml-1) did not differ (p greater than 0.05) from serum free levels (x = 0.251 +/- 0.139 microgram ml-1), supporting the concept of the synovial cavity as a compartment readily accessible to unbound drug species. Multiple regression analyses revealed relative independence of free concentration on total levels, and the small positive and negative influences of NEFAs and albumin concentrations. Bilirubin, uric acid, cholesterol, triglycerides, and chloride ion did not correlate with serum free concentration. PMID- 7471615 TI - Influence of DH/DL alleles regulating debrisoquine oxidation on phenytoin hydroxylation. AB - Eleven subjects of previously determined debrisoquine oxidation phenotype status (extensive metabolizer [EM], n = 5; poor metabolizer [PM], n = 6) were studied for their ability to perform the aromatic 4-hydroxylation of phenytoin. The PM subjects studied were found to be slower metabolizers of phenytoin than EM subjects in terms of the metabolite formation rate constant (kfHPPH: EM, 0.030 +/ .007 hr-1; PM, 0.016 +/- 0.003 hr-1, 2p less than 0.001) and cumulative excretion of 4-hydroxyphenytoin (48 hr after dosing: EM, 52.8 +/- 10.7%; PM, 36.9 +/- 7.0%, 2p less than 0.01). It is concluded that the metabolic oxidation of phenytoin is influenced by the same DH and DL alleles, acting at the same locus, that regulate the hydroxylation of debrisoquine and that impaired metabolism of phenytoin may be expected to occur in about 9% of the population, being transmitted as an autosomal-recessive trait. It is suggested that debrisoquine oxidation phenotyping may have predictive value in guiding phenytoin dosage, particularly in those with impaired oxidation. PMID- 7471616 TI - Early hemodynamic and humoral effects of lofexidine. AB - Hemodynamic and humoral effects of lofexidine were assessed in 11 patients with essential hypertension after a total of 1.5 mg were given orally over 24 hr. Heart rate (bpm) slowed (-12 +/- 6 [SEM], p less than 0.05) and cardiac output (liters per minute) was reduced (-0.78 +/-0.18, p less than 0.01) irrespective of blood pressure response; the latter was related to changes in systemic resistance (TPR) (r = 0.72, p less than 0.01). Cardiac performance judged from ejection fraction (0.61 +/- 0.03 to 0.59 +/- 0.03, NS) and mean transit time (8.73 +/- 0.53 sec to 9.20 +/- 0.35, NS) were not altered. Plasma volume was expanded more than 10% in two patients but not changed in the others. Supine plasma catecholamines determined in five patients were reduced in all but one with no correlation to changes in either TPR or diastolic blood pressure. On the other hand, there was an increase in plasma catecholamines during head-up tilt in four of five patients, indicating normal catecholamines release. Orthostatic hypotension occurred de novo in three patients; two of them had simultaneous slowing of heart rate (vasovagal attack). Results suggested that reduction of blood pressure by lofexidine depended on lack of increase in TPR in response to reduction of cardiac output; the hemodynamic pattern of this centrally acting adrenergic blocker closely resembled that reported for beta blockers. PMID- 7471617 TI - Bronchodilation and inhibition of induced asthma by adrenergic agonists. AB - In asthma, adrenergic agonists alleviate airflow obstruction and prevent obstructive responses to a variety of stimuli. A rapidly and a slowly metabolized agonist were compared to determine whether bronchodilation is the major mechanism by which these drugs prevent exercise-induced asthma (EIA). A 200-microgram inhaled dose of the rapidly metabolized agonist, isoproterenol, induced bronchodilation of the same order as terbutaline 500 microgram (1-sec forced expiratory volume [FEV1] increased 9.5% and 10.2%). An hour after isoproterenol, FEV1 was still above baseline (p less than 0.02) but EIA was only partially inhibited; the 23% fall in FEV1 was of the same order as the 32% fall after placebo (p greater than 0.05). One hour after terbutaline, mean resting FEV1 was in the range of that after isoproterenol, but the 10% change after exercise was less than that after placebo and isoproterenol (p less than 0.005). Our findings suggest that the two effects have different dose-response relationships, with higher doses of adrenergic agonists needed to prevent EIA than to maintain bronchodilation. PMID- 7471618 TI - Kinetic equivalence of stable-isotope-labeled and unlabeled phenytoin. AB - Stable isotope labeling (SIL) of a drug results in a higher molecular weight than that of the unlabeled drug. SIL tracer doses can be quantitated separately from unlabeled drug by gas chromatography-mass spectrometry (GC-MS) without exposing the patient to radiation. The higher molecular weight of SIL drug could cause a higher energy of activation for (and slowing of) metabolic reactions ("isotope effect"). To evaluate possible isotope effect, three dogs and three men were infused with a mixture containing equal amounts of SIL (2-13C-1,3-15N2) and unlabeled phenytoin (PHT). Plasma and urine were collected at regular intervals. Concentrations of SIL and unlabeled PHT and HPPH (the major metabolite of PHT) were determined by GC-MS. Within each subject there was no trend for concentrations of SIL PHT or HPPH to be higher or lower than concentrations of their unlabeled analogs (greater than 0.20 to 0.90). There was no difference in the distribution and elimination half-lifes (t 1/2s), volume of distribution, volume of central compartment, or clearance of the two forms of PHT. Thus, no isotope effect was found. PMID- 7471619 TI - Influence of range of renal function and liver disease on predictability of creatinine clearance. AB - Several formulas for predicting creatinine clearance (Ccr) are used for adjusting drug dosages but limited data are available on their accuracy in patients with significant renal impairment or concurrent disease. We measured 144 Ccr in 103 patients and compared results using four predictive methods. Of nine common diseases in these patients, liver disease was associated with a large (p less than 0.02) prediction error (overprediction). After data from eight patients with liver disease were removed, there was good overall correlation between predicted and measured Ccr (r2 = 0.91 for each method) but only two of the methods (I and IV) were consistently accurate in all ranges of renal function. Methods for predicting Ccr should not be used in patients with liver disease. PMID- 7471620 TI - Changes in plasma drug binding and alpha 1-acid glycoprotein in mother and newborn infant. AB - A number of drugs bind to alpha 1-acid glycoprotein in plasma. To determine whether age-related changes in alpha 1-acid glycoprotein influence drug binding in mother and newborn infant and also the effects of sex, pregnancy, and oral contraceptives on drug binding, the binding of lidocaine, diazepam, propranolol, d-tubocurarine, and metocurine was determined by equilibrium dialysis in 17 men, 16 nonpregnant women, 16 nonpregnant women on oral contraceptives, and 15 mothers and their newborn infants at delivery. The free fraction of d-tubocurarine (p less than 0.05), metocurine (p less than 0.01), propranolol (p less than 0.001), and lidocaine (p less than 0.02) was higher in neonatal blood than in maternal blood, whereas the free fraction of diazepam was less in fetal cord blood than the mother's (p less than 0.02), but higher than that in nonpregnant women (p less than 0.001). The free fractions of diazepam, propranolol, and lidocaine were higher (p less than 0.001) in pregnant women compared with nonpregnant women while the diazepam and lidocaine free fractions were higher (p less than 0.05) in nonpregnant women and oral contraceptives than in women not using them. Alpha 1 acid glycoprotein was lower in the fetus (15.3 +/- 4.7 mg/100 ml) than the mother (49.6 +/- 6.5 mg/100 ml) (p less than 0.002). There was a positive correlation between plasma alpha 1-acid glycoprotein concentrations and the binding ratio (bound/free concentrations) of lidocaine (p less than 0.001; r = 0.623) and propranolol (p less than 0.001); r = 0.652), indicating that it is likely that the elevation of the free fraction of these drugs in the fetus is due in part to lower levels of alpha 1-acid glycoprotein. PMID- 7471621 TI - Methapyrilene kinetics and dynamics. AB - A study was undertaken to characterize the H1 receptor blockade, central nervous system depressant properties, and kinetic parameters of methapyrilene in man. Eight healthy subjects received, in random order at weekly intervals, placebo and methapyrilene 20 mg intravenously and 50 mg and 25 mg orally. Methapyrilene exhibited a moderate antihistaminic effect as measured by the reduction of histamine-provoked skin wheals. Sedation and drowsiness were detected only at the first sampling time (0.75 hr) after intravenous doses. The terminal plasma half life ranged from 1.1 to 2.1 hr, apparent volume of distribution from 2.14 to 6.61 1/kg, and plasma clearance from 0.013 to 0.048 1/min/kg. Systemic bioavailability was low and showed large interindividual differences, ranging from 4% to 46%. Recovery of unchanged drug from the 24-hr urine was under 2% of the doses. PMID- 7471623 TI - Clinical biostatistics. LV. The t test and the basic ethos of parametric statistical inference (Part I). PMID- 7471622 TI - The method of separate exponentials: a simple aid to devising intravenous drug loading regimens. AB - Stimulation of complex dosage regimens for drugs with multicompartmental kinetics is described using the method of separate exponentials. This approach requires that alpha- and beta-phases are treated separately throughout and summed only at the end of the stimulation. The method was used to devise a loading regimen for pirmenol, comprising a priming injection, and a rapid loading infusion, followed by a maintenance infusion. The regimen was tested in a patient with excellent agreement. The method of separate exponentials is mathematically simple and of informational value in that it demonstrates when the early distribution phase is important. Its use can avoid the potentially dangerous assumption of one compartmental kinetics in the design of intravenous loading regimens. PMID- 7471624 TI - The clinical and radiological features of the empty sella syndrome. AB - The importance of the empty sella syndrome has increased with recent interest in pituitary micro-adenomas, trans-sphenoidal hypophysectomy and computerised tomography. Four cases of the empty sella syndrome which illustrate the clinical and radiological features are reported, and the literature is reviewed. Headache, visual symptoms and endocrine disturbances are commonly found in association with the empty sella syndrome, and the coexistence of pituitary micro-adenomas is well recognised. Symmetrical ballooning of the sella turcica without changes in configuration is the typical finding with plain radiography, but occasionally more extensive destructive changes in the region of the pituitary fossa may occur. Provided that the correct angulation, collimation and level of cut are chosen, computerised tomography now may be used to make a definitive diagnosis, making air encephalography unnecessary. PMID- 7471625 TI - Computed tomography (CT) in parenchymatous cerebral cysticercosis. AB - Cysticercosis is an infection with a larval or a bladder-worm stage of the species of Taenia. Cysticerci have a predeliction for the nervous system where they may be found in the meninges, the ventricles and within the brain itself. Brain involvement or parenchymatous form has an acute and chronic phase. Before the advent of CT scanning radiology was of little value in the acute parenchymatous cysticercosis, but with CT the changes within the brain can be recognised. In reviewing 14 cases of acute parenchymatous cysticercosis three CT patterns were found. 1. A diffuse low density pattern with no or little change after contrast medium 2. Multiple low densities with small rounded central areas of enhancement. 3. Large cystic lesions which may become ring lesions after contrast medium. We conclude that in an endemic area for cysticercosis, when one of these CT patterns is present, cysticercosis should be considered in the differential diagnosis. In a child where the clinical features are suggestive, the CT pattern may be diagnostic of cysticercosis. In the chronic parenchymatous stage the cysts have calcified and this may occur within one year of the acute onset. In 11 cases of chronic parenchymatous cysticercosis where the calcification was visible on the plain skull radiographs the CT scan added very little additional information. It may however be of value in confirming intracranial calcifications where their presence on the plain skull radiograph is in doubt. PMID- 7471626 TI - The role of computed tomography in tumours of the larynx. AB - A clinical study of computed tomography (CT) was undertaken in 35 patients with tumours involving the larynx. Twenty-seven had primary laryngeal neoplasms, five had tumours arising in adjacent structures but invading the larynx and three who had undergone total laryngectomy were investigated for possible recurrence. The findings were compared with conventional radiological methods and clinical assessment. Confirmation was obtained from laryngectomy specimens in four patients and at autopsy in one. CT provided additional preoperative information on 14 occasions. This included better delineation of submucosal tumour extent, invasion of the pre-epiglottic space and cartilage displacement in invasion. PMID- 7471627 TI - Magnification lymphography. AB - Magnification lymphography was performed on 37 patients, 22 of whom were suffering from bladder carcinoma, and compared with non-magnification lymphograms by two observers independently. The magnification lymphograms were performed on a Siemens Microfocus Bi 125/3/50 RG tube with a 0.1 mm focal spot. In six of the 37 patients (16%) significant extra information was found on the magnification lymphograms compared to the non-magnification lymphograms. In 31 out of 37 patients (84%), the intranodal detail shown was better in the magnification radiographs. Against the additional information gained must be weighed the disadvantages of magnification lymphography which include increased radiation dose, lengthening of procedure time, increased cost and the critical nature of exposure factors. In conclusion, magnification lymphography has a place and the disadvantages seem to be outweighed by the advantages. PMID- 7471628 TI - Changes in lymph node size induced by lymphography. AB - Although the contraction of normal lymph nodes after lymphography has been measured the enlargment caused by lymphography has not. In this study the lengths and widths of 25 radiologically normal nodes were measured in five patients who had a second lymphogram carried out when these nodes were still outlined by contrast medium from the first lymphogram. If the mean length after the second lymphogram is taken as 100% then the mean length 24 h earlier measured 82%, and if the mean volume after the second lymphogram is taken as 100% then the mean volume 24 h earlier was 49%. These substantial increases are similar to the reductions which have been reported in some previous papers and are probably due to distension of lymph node sinuses by oily contrast medium. It is obviously important that changes in lymph node size which are induced by lymphography itself be taken into account when follow-up films and repeat lymphograms are used to assess the response of lymph nodes to treatment. PMID- 7471629 TI - Changes in the size of pelvic lymph nodes in patients being followed after radiotherapy for Hodgkin's disease. AB - Serial abdominal radiographs of 10 patients who had shown fluctuation in the size of opacified pelvic lymph nodes were reviewed. All had previously been treated for Hodgkin's disease by irradiation alone. Three of the patients studies were finally diagnosed as having recurrent Hodgkin's disease, but the other seven were diagnosed as having reactive change and have remained well without evidence of relapse in the pelvis or abdomen after four years follow-up; one of these patients received quadruple chemotherapy for recurrent disease at the presenting site in the neck. The differential diagnosis of enlarging pelvic lymph nodes in irradiated Hodgkin's patients may be difficult and recurrent lymphoma should not be assumed. Radiologically, reactive change is favoured by the simultaneous enlargement of several groups of nodes, whereas recurrence is favoured by the selective increase in size of solitary or neighbouring lymph nodes. In case of doubt, cytological or histological confirmation of relapse should be sought. PMID- 7471630 TI - Lymphography in the evaluation of urinary bladder carcinoma. AB - Lymphography was performed in 41 patients with bladder carcinoma. The lymphographic findings were correlated to the clinical and histological staging and the pre-operative grading. Twenty-seven patients were operated upon. The findings of lymphography were compared with the findings at operation, and the pathological examination of the removed lymph nodes. The lymphographic diagnosis was proved to be correct in 77.7% of the patients. There were two false positive lymphographies, and one false negative. Lymphography in groups T3 or T4 yielded a high diagnostic accuracy. PMID- 7471631 TI - Bipedal lymphography in the management of bladder cancer. AB - Bipedal lymphography was performed in 91 patients with transitional cell carcinoma of the bladder. Equivocal reports were submitted in 18 (19.8%) examinations. In this series 18 (19.8%) of the patients died within one year of first treatment by radical radiotherapy. The prognosis was poorer in patients with more locally advanced tumours, in whom there was a higher proportion of abnormal lymphograms. An abnormal lymphogram was also related to a poorer prognosis than if the lymphogram was normal. This difference was greater when associated with a poorly differentiated cancer. The implications for management are discussed. PMID- 7471632 TI - Internal mammary lymphoscintigraphy in the conservative surgical management of breast carcinoma. AB - Radiocolloid internal mammary lymphoscintigrams were carried out in 249 women who had undergone partial mastectomy. Only 39 patients, or 15%, had axillary sampling; of these, 19 were positive. At last follow-up, a minimum of two years from the date of study, the overall recurrence rate was 67% for patients with an abnormal lymphoscintigram and 34% for those with a normal lymphoscintigram. Actuarial survival rates projected to five years indicate a statistically significant difference in the prognostic implications of the normal and abnormal internal mammary lymphoscintigram, with 83 and 59% survival rates, respectively. PMID- 7471633 TI - An investigation of radiological, clinical and pathological correlations in osteoarthrosis of the hip. AB - Seventy per cent of cases of osteoarthrosis of the hip are regarded as being 'idiopathic' in type. Idiopathic osteoarthrosis of the hip is twice as common in females as in males. Osteoarthrosis of the hands shows a definite association with idiopathic osteoarthrosis of the hip, especially in women. Osteoarthrosis of the knees is found in over half the cases of idiopathic osteoarthrosis of the hip and of the hands, although there are sex differences in the distribution of hand and knee osteoarthrosis in these cases. Hand osteoarthrosis is not related to hand laterality, although osteoarthritic changes are likely to be more severe in the dominant hand when there is osteoarthrosis of both hands. No convincing evidence was found of a relationship between the various slab radiograph patterns from femoral heads and idiopathic osteoarthrosis of the hip. There was no definite association between idiopathic osteoarthrosis and the presence of various tissue antigens. The identification of a group of patients with idiopathic osteoarthrosis suggests a generalised cartilage abnormality, with the development of osteoarthrosis being possible related to subsequent stress changes. PMID- 7471634 TI - Symptomatic vertebral haemangiomas. AB - Twelve cases of symptomatic vertebral haemangioma are reported in respect of their clinical presentation and radiological findings. One patient was only 10 years old, the youngest patient ever reported. Less commonly known findings of associated paravertebral mass and the displacement of the thoracic paraspinal line are discussed. PMID- 7471635 TI - Unusual manifestations of neonatal pharyngeal perforation. AB - The small series of previously described cases of neonatal pharyngeal perforation are reviewed and two additional cases with atypical initial presentations are described. The condition is more common than is currently appreciated, as suggested by one of our cases which remained undetected until many years later. We also wish to emphasise the high location of perforation despite an initial presentation which may appear to the contrary. PMID- 7471636 TI - Barium enema in suspected necrotising enterocolitis: is it ever indicated? AB - Three infants with suspected, and subsequently proven, necrotising enterocolitis (NEC) had contrast medium enemas early in the course of their disease to assist diagnosis. There were no adverse effects from the enemas. All patients had evidence of persistent spasm of segments of the colon and contour speculation, probably representing superficial mucosal ulceration. It is suggested that when early or mild forms of NEC present diagnostic problems, a carefully performed contrast enema may be of diagnostic value. Absence of deep transmucosal necrosis in such cases definitely decreases the risk of perforation during the examination. PMID- 7471637 TI - Crohn's disease of the small intestine: a comparative study of the radiological and pathological appearances. AB - A prospective study was carried out to compare the radiological and pathological appearances of Crohn's disease of the small intestine. The X-ray examinations were carried out by infusing barium directly into the small intestine. Thirty-six patients who were operated on for Crohn's disease of the small intestine are included. There was excellent correlation between the radiological appearances and the findings at morbid anatomy. PMID- 7471638 TI - Crohn's disease of the small intestine: a review of the radiological appearances in 100 consecutive patients examined by a barium infusion technique. AB - The radiological features of Crohn's disease of the small intestine are described in a report on 100 patients examined by a barium infusion method. The examination is performed by introducing a large volume of barium suspension through a tube directly into the small intestine. A multiplicity of radiological signs were seen in the majority of the examinations. Discrete and fissure ulcers were present in many cases; longitudinal ulcers, sinuses and fistulae were seen less frequently. Other signs commonly seen were strictures, in many cases with proximal dilatation, thickening and distortion of the mucosal folds, cobblestoning, asymmetrical involvement, thickening of the wall of diseased intestine and good demarcation of normal from abnormal small intestine. Adhesions, skip lesions, gross distortion, a featureless outline of the diseased intestine and pseudo polyps were less commonly seen. The disease was more extensive in patients who previously had resections for small intestinal Crohn's disease. PMID- 7471639 TI - The influence of emergency urography and haematuria on the diagnosis of ureteric colic. AB - The literature concerning the investigation of ureteric colic is reviewed. I have compared previous papers with the results of a prospective study of 50 patients who underwent emergency urography and testing for haematuria. Haematuria was present in 88% of all patients and in 97% of those with proven ureteric colic. The high haematuria figure is thought to be due to patient selection. Thirty-four patients (68%) had abnormal urographic findings consistent with ureteric colic. Seven of these patients (20%) had no plain film abnormality and the contrast films therefore confirmed the diagnosis. Haematuria is a useful indication of ureteric colic. However, since ureteric obstruction occurs without haematuria, and a normal urogram may be associated with haematuria, emergency urography is essential to confirm ureteric colic. This should be performed soon after the onset of pain if maximal information is to be obtained, thereby avoiding unnecessary investigation. PMID- 7471640 TI - Percutaneous aspiration biopsy of abdomen and retroperitoneum. AB - Percutaneous fine needle aspiration biopsy was performed in 38 patients. Adequate material for histological diagnosis was obtained in all. No false positive results were obtained and there was only one false negative result. The procedure was without complication in all patients. The biopsy material was obtained by the radiologist in the X-ray department following localisation of the lesion by ultrasound. The advantages of the early use of this technique in the evaluation of patients presenting with suspected mass lesions in the abdomen and retroperitoneum are described, and its contribution to the formulation of further patient management stressed. PMID- 7471641 TI - The effect of an energy-restricted diet on the protein metabolism of obese adolescents: nitrogen-balance and whole-body nitrogen turnover. AB - 1. The effect of an energy-restricted (46 kJ day-1 kg-1), adequate protein diet (1.47 g day-1 kg-1) on the nitrogen metabolism of five obese rapidly growing adolescents (two males and three females) was assessed by means of nitrogen balance measurements and determination of whole-body nitrogen turnover. 2. The energy-restricted diet had no significant effect on nitrogen balance (P more than 0.1) for the entire group when compared with the control dietary intake; however, significant (P less than 0.01) differences in nitrogen balance were noted among individuals at each dietary interval. 3. Mean values for whole-body nitrogen turnover, protein synthesis and breakdown for the control period were: 45.5 +/- 13.2 mg of nitrogen day-1 kg-1, and 5.72 +/- 1.96 and 5.74 +/- 1.92 g of protein day-1 kg-1 respectively. These values are 82% of those measured in infants. 4. Reducing the mean non-protein energy intake to 20 kJ day-1 kg-1 had no significant effect on whole-body nitrogen turnover, protein syn;thesis or protein breakdown. 5. The results are discussed in relation to the regulation of whole body nitrogen metabolism, by dietary protein and energy intakes. PMID- 7471642 TI - Metabolic aspects of hypothermia in the elderly. AB - 1. Plasma concentrations of glucose, lactate, amino acids, non-esterified fatty acids, glycerol, ketone bodies, insulin and cortisol were measured in 43 elderly patients with hypothermia. In 15 of these patients forearm arteriovenous differences were also measured. Core temperatures ranged from 25.9 to 35.5 degrees C. 2. The metabolic state was of mobilization of glycogen and triacylglycerol stores, with high plasma concentrations of lactate and lipid metabolites. The plasma concentration of glucose was raised in those with hypothermia of a short duration (less than 6 h). In other patients it was low in those with core temperatures around 30 degrees C, but below this temperature it was variable and often high. Concentrations of other metabolites or hormones were not related to core temperature. 3. Plasma concentrations of cortisol were high and positively correlated with those of lactate and glycerol, suggesting active involvement in stimulation of muscle glycogenolysis and of lipolysis. 4. Plasma concentrations of insulin ranged from very low to very high and appeared to depend on the concentrations of both glucose and alanine. 5. Arteriovenous differences were generally small. There was peripheral release of lactate and of amino acids but no overall peripheral uptake of glucose. In nine out of 15 patients there was a significant peripheral release of glucose. PMID- 7471643 TI - Pathways of transepithelial potassium movement in the epithelium of distal colon in man. AB - 1. Methods for studying the kinetics of movement of radioactive tracers across human distal colonic epithelium have been developed for use in vivo and applied to examination of transepithelial transfers of potassium. 2. During absorption of 43K from the lumen, two potassium pools, A and B, in the epithelium were identified, having mean turnover half-times of 16 and 87 min respectively. On average about 50% of the potassium lumen-to-plasma flux and most, if not all, of the potassium plasma-to-lumen flux took place through pool A. The transcellular pathway (equated with pool B) appeared to be of relatively little significance in transepithelial movement of potassium in the human distal colon. 3. Sodium and chloride movements were unaffected when lithium was present in the luminal solution, but the potassium secretion rate was nearly trebled due to increased plasma-to-lumen flux through the fast turnover pool A. 4. The permeability of the epithelium to potassium, as judged by the plasma-to-lumen fluxes, greatly exceeded that to sodium. This appeared to depend on a special highly selective potassium channel which differed from the shunt pathway in possessing definite capacity. PMID- 7471644 TI - Absorption of artificial effusions from synovial joints: an experimental study in rabbits. PMID- 7471645 TI - Interaction of conventional and antikaliuretic diuretics with the renal prostaglandin system. PMID- 7471646 TI - Total joint replacement arthroplasty: review of current literature. PMID- 7471648 TI - [Pediatric surgery: considerations and clinical findings concerning the postoperative use of a "biological reconstituent"]. PMID- 7471647 TI - [Alcoholism within a medical department. I. Clinico-statistical study]. PMID- 7471649 TI - [Hepatic activity of (+)-cyanidanol-3 in animal experiments]. PMID- 7471650 TI - [Therapeutic aspects of alcoholism in a medical department. II]. PMID- 7471651 TI - [Current diagnostic and therapeutic trends in obliterating arteriopathies: oscillometry and Doppler examination]. PMID- 7471652 TI - [Silymarin in the prevention of hepatic damage by psychopharmacologic drugs. Experimental premises and clinical evaluations]. PMID- 7471653 TI - [Methergoline in blocking of post-partum lactation]. PMID- 7471654 TI - [Use of a new posterior pituitary derivative (ornithine-8-vasopressin) with vasoconstrictive action in gynecologic surgery]. PMID- 7471655 TI - [List of prescriptions for therapy of acute pulmonary edema]. PMID- 7471656 TI - [Kinetics of BSP disappearance after a single dose of cimetidine in ulcer patients with liver disease]. PMID- 7471657 TI - [Photoplethysmographic changes after the cold pressure test in basal conditions and after oral administration of nitrates in arterial disease and control subjects]. PMID- 7471658 TI - [Extracts of adrenal cortex and their relation with the function of the pituitary adrenal axis and blood glucose regulation in elderly subjects]. PMID- 7471659 TI - [Reflections on 26 cases of tubercular meningitis]. PMID- 7471660 TI - [Case of hypogammaglobulinemia: clinical and immunological study, and possibilities of therapy]. PMID- 7471661 TI - [Citrobacter freundii osteomyelitis of the mandible: review of the literature and report of a case]. PMID- 7471662 TI - Acetaminophen pharmacokinetics after overdose. AB - Concentrations of acetaminophen in serum and urinary excretion rates of acetaminophen glucuronide and acetaminophen sulfate were determined in a 15-year old female who had ingested an overdose which resulted in the absorption of an estimated 9.92 g of acetaminophen. The results obtained are in reasonably good agreement with predictions of acetaminophen disposition based upon a previously developed pharmacokinetic model of capacity-limited acetaminophen elimination, but additional studies are needed to refine that model. PMID- 7471664 TI - Fragility of erythrocytes in animals affected by lantana poisoning. AB - Clinical observations on four male cow calves and three other lantana poisoned animals under field conditions have been made along with hematological studies, including observations on plasma bilirubin content and osmotic fragility of erythrocytes. Four healthy male cow calves served as control. Five clinically advanced cases succumbed within 1 to 10 days of observation. Hematocrit values in four of the seven affected animals were abnormally high. The total plasma bilirubin content in the fatal cases was of the order of 5 to 50 times the normal value while, in calves that recovered, it was not more than 3 times. The osmotic fragility of erythrocytes was measured by the rate of hemolysis in decreasing concentrations of saline. The erythrocytes of normal animals resisted hemolysis till the saline concentration was lowered to 0.60%, and it was complete at the low saline concentration of 0.30%. However, in lantana poisoned animals, hemolysis started even in 0.72% saline and was complete at a saline concentration of 0.44%. The hemolysis curves of animals that died were far removed from the normal, but those of the calves that recovered and the one that died but had the lowest bilirubin content closely approached the normal curve. PMID- 7471663 TI - Clinical findings, elimination pharmacokinetics, and tissue drug concentrations following a fatal amitriptyline intoxication. PMID- 7471665 TI - Mass screening and confirmation of phencyclidine (PCP) in urine by radioimmunoassay/TLC. AB - A sensitive, rapid, and specific procedure is described for the mass screening and confirmation of PCP (phencyclidine) in urine specimens at concentrations as low as 0.3 microgram of PCP per mL. Specimens that test positive by radioimmunoassay must be confirmed by TLC using methylene chloride:n butanol:concentrated NH4OH (85:15:0.5) as the developing solvent system. This separates PCP from other drugs or endogenous urinary substances. PMID- 7471666 TI - What young children think you see when their eyes are closed. PMID- 7471667 TI - A deletion ahead of its time. PMID- 7471668 TI - Is the human sentence parsing mechanism an ATN? PMID- 7471669 TI - Childhood croup. PMID- 7471670 TI - Growth disorders in children. PMID- 7471671 TI - Health problems of adolescents. PMID- 7471672 TI - Avoiding unnecessary X-ray exposure in children. PMID- 7471673 TI - Infantile colic, or what to do until the fourth month comes. PMID- 7471674 TI - Anemia due to bone marrow failure: diagnosis and treatment. AB - In the last decade a large increase of our basic understanding concerning erythropoietin and the regulation of erythropoiesis has led to improved methods for the cell culture of human bone marrow cells. These culture methods in turn have been applied to bone marrow failures with a remarkable increase in our knowledge of the pathogenesis of some of these conditions, particularly the aplasias. The pathogenesis of pure red cell aplasia has been elucidated, and 60% of these patients have been shown to respond to cytotoxic, immunosuppressive treatment. Bone marrow transplantation has proved to be very helpful in the treatment of aplastic anemia and has provided impetus for increased knowledge concerning the pathogenesis of the aplasia. Some of these patients may have suppression of marrow hematopoiesis by the marrow T-cells and can be successfully treated with antilymphocyte globulin or high-dose prednisolone. The future looks bright for further clinical advances concerning the bone marrow failures, but more must be learned about the pathogenesis of these anemias if improved methods of treatment are to be developed. PMID- 7471675 TI - Polycythemia: differential diagnosis and treatment. PMID- 7471676 TI - Iron deficiency in clinical practice. PMID- 7471677 TI - The practicing physician in modern medicine. PMID- 7471678 TI - Hemolytic anemia: diagnosis and management. PMID- 7471680 TI - Human erythrocyte G6PD deficiency: pathophysiology, prevalence, diagnosis, and management. PMID- 7471679 TI - Sickle cell disease. PMID- 7471681 TI - Diagnosis, staging, and treatment of the malignant lymphomas. PMID- 7471682 TI - Gaucher's disease. PMID- 7471684 TI - Measurement of cardio-respiratory fitness and body composition in the clinical setting. PMID- 7471683 TI - The effect of aging on the blood count. AB - There are age-related changes in the results of some tests, but in none is the normal value for the elderly sufficiently different from normal values for younger adults to permit the physician confidently to ascribe an abnormal result to age alone. In assessing the significance of abnormal results of hematologic tests in asymptomatic patients, it is particularly important to consider problems of major import, such as blood loss from the gastrointestinal tract. Anemia is the abnormality most likely to be encountered, and investigation of a marginal reduction in the hemoglobin level of an elderly patient may be rewarded with a specific diagnosis of a disease that can be effectively treated, at least for a time. PMID- 7471685 TI - Exercise programs for the cardiac patient. PMID- 7471686 TI - Common athletic injuries in women. AB - Overwhelming clinical experience with injuries in female athletes suggests that a slow and progressive training program should be used for the novice female athlete, with particular emphasis on increasing flexibility and strength. Females may expect to engage actively in competitive or noncompetitive athletics with significant benefits of personal well-being and improved self image. Early recognition of the problems in athletically active females should improve management and hasten return to the desired functional level. PMID- 7471687 TI - Exercise and children. AB - Children are naturally active and enjoy many forms of physical activity. Each community, family, and physician owes the children in his care the opportunity to exercise and compete to the limits of their ability under supervised programs that will encourage a life-long participation. PMID- 7471688 TI - The spine in sports. PMID- 7471690 TI - Allergic and irritant contact dermatitis. PMID- 7471689 TI - Sports injuries and arthritis. PMID- 7471691 TI - Skin disorders in the elderly. PMID- 7471692 TI - Cutaneous fungal infections. PMID- 7471693 TI - Adverse reactions to sunlight. PMID- 7471694 TI - The pruritic dermatoses. AB - Systemic diseases that produce itching are listed in Table 1. Control of prurtis in each case depends on control of the fundamental disease. Intolerable itching may require the combined services of a generalist, a dermatologist, an internist or subspecialist. Prurtis is a common symptom, not completely understood, mediated by stimulation of nerve endings in the skin. It may be produced by internal or external stimuli, may be mild or severe, and its cause is often obscure. Treat the symptoms and watch for developments. PMID- 7471695 TI - Early recognition of cutaneous neoplasms. PMID- 7471696 TI - Topical steroids in the treatment of common dermatoses. PMID- 7471697 TI - Verb semantic structures in memory for sentences evidence for componential representation. PMID- 7471698 TI - Serving the mental health needs of the elderly: the case for removing barriers and improving service integration. AB - Utilization rate information reveals that elderly persons are not adequately served by the existing mental health service system. The authors identify cultural, systems, and resource related barriers to full service, emphasizing psychological barriers which may be present in both the elderly client and the therapist. Overcoming these barriers will require, first, that professional training programs and local in-service training activities be modified to expand multi-disciplinary learning opportunities. Then, the authors advocate systems change to move through in-house outreach, advocacy and programming activities toward an integrated system of community services for the elderly. Mental health workers are seen as uniquely qualified to serve as standard-bearers in developing a locally integrated service delivery system. PMID- 7471699 TI - Community mental health centers and the elderly: a time for expansion of planning, research, and demonstration projects. AB - The purpose of this paper is to acquaint persons working in Community Mental Health Centers with federal funding resources which support planning, research, and demonstration projects related to mental health and aging. The significance of exploration by CMHCs into the aging area is suggested by the legislative histories of CMHCs and aging programs, our expanding aging population which requires increasing health and mental health services, and the growing number of resources available for the development of services related to aging. Principal funding sources related to mental health and aging are: Alcohol, Drug Abuse and Mental Health Administration, National Institutes of Health; Administration on Aging; and Health Care Financing Administration. Each funding source is presented through a review of five key items: 1) name of program; 2) objectives; 3) funds awarded; 4) legislative mandate; and 5) contact person and address. Program areas for each funding source are discussed. Knowledge of these funding resources should enable CMHCs to plan services and to expand capabilities to serve the growing over-60 population. PMID- 7471700 TI - "Emergency room" and "mobile response" models of emergency psychiatric services. AB - The present study surveyed federally-funded emergency psychiatric programs in 33 California community mental health centers (CMHCs). Four aspects of service organization were examined: (1) the availability of various response styles (telephone service, walk-in service, follow-up home visits, mobile initial response); (2) the types of staff providing the service; (3) the location of the programs, and (4) the organizational setting of the CMHCs (consortium or single agency centers). From these characteristics of service organization two contrasting types of emergency psychiatric programs were derived--the "emergency room" and "mobile response" models. The relationship between these two models of service delivery and catchment area demographic characteristics was examined and no remarkable trends emerged. The implications of developing a typology of emergency psychiatric programs are discussed with respect to a scheme for evaluating the relative effectiveness of services and for planning future programs. PMID- 7471701 TI - Salient hopes and fears: social marketing to promote human services. AB - The application of social marketing principles to the promotion of human services programs is illustrated in this paper. As part of a household interview survey, 176 respondents were asked to name their hopes and fears for their lives. Responses generated by these questions are reported briefly. These findings then are interpreted from the perspective of social marketing theory, and resulting implications for the design of strategies to promote human service programs are discussed. Other areas in which social marketing theory may aid the planning and delivery of human services are mentioned. PMID- 7471702 TI - Toward a clarification of primary prevention. AB - The ambiguity of the concept of primary prevention and a vague fear that primary prevention may violate civil liberties are two of the important barriers blocking the implementation of primary prevention. Progress toward overcoming these barriers may be possible by distinguishing between the different but overlapping meanings of primary prevention. In this paper four models of primary prevention are proposed to aid definitional accuracy and to help identify and avoid those types of preventive interventions which threaten civil liberties. PMID- 7471703 TI - Another role for citizens: three variations of citizen evaluation review. PMID- 7471704 TI - [Comparison of influenza antibody serologic immunity in man and in pig (author's transl)]. PMID- 7471705 TI - [Surveillance of influenza virus in healthy pigs (author's transl)]. PMID- 7471706 TI - [Evaluation of an inactivated, adjuvated vaccine against swine influenza (author's transl)]. PMID- 7471707 TI - Isolation of influenza viruses from avian species in Great Britain. PMID- 7471708 TI - [Isolation of Hav6 N2 influenza virus from diseased laying chickens (author's transl)]. PMID- 7471709 TI - [Circulation of influenza viruses in the bay of the Somme river (author's transl)]. PMID- 7471710 TI - Characterization of avian influenza viruses isolated in Israel in 1978-1979. PMID- 7471711 TI - Report on avian influenza outbreak in Minnesota turkeys--1978. PMID- 7471712 TI - Serological response to whole, split and subunit influenza vaccines of persons with and without immunological experience towards influenza A/U.S.S.R. 90/77 virus. PMID- 7471713 TI - Clinical study of a trivalent subunit influenza vaccine containing antigens of the (H1N1)-subtype. PMID- 7471714 TI - Studies on influenza virus isolated from migrating birds in Egypt. PMID- 7471715 TI - Antigenic analysis of influenza A virus surface antigens: considerations for the nomenclature of influenza virus. PMID- 7471716 TI - Antiphagocytic factors of Staphylococcus aureus. PMID- 7471717 TI - Immunity to chlamydial infections in mice--I. Establishment of quantitative techniques. PMID- 7471718 TI - Immunity to chlamydial infections in mice--II. A quantitative study of respiratory challenge. PMID- 7471719 TI - Alternate complement pathway in porcine sera: lysis of guinea pig erythrocytes. PMID- 7471720 TI - DEAE cellulose chromatographic characterization of canine serum. PMID- 7471721 TI - Generation of temperature profiles for elements of the human body. PMID- 7471722 TI - A computer method for cardiac radionuclide kymography. PMID- 7471723 TI - A unique versatile, real-time software package for controlling a Nicolet 1074 averager from a laboratory minicomputer. PMID- 7471724 TI - Human chromosome classification using discriminant analysis and Bayesian probability. PMID- 7471725 TI - An interactive, microcomputer-based karyotype analysis system for phylogenetic cytotaxonomy. PMID- 7471726 TI - Anthandbone--a program for following the growth of children by comparison with standards. PMID- 7471727 TI - A necessary and sufficient condition of global structural identifiability of compartmental models. PMID- 7471728 TI - Caring for the whole patient: health advocacy. PMID- 7471729 TI - The "exceptions" to informed consent (second of two parts). PMID- 7471731 TI - Nothing but problem solving? PMID- 7471730 TI - Study of health maintenance organizations. AMA Council on Medical Service. PMID- 7471732 TI - Evaluation of neck mass: computed tomography. PMID- 7471733 TI - Current concepts in the surgical treatment of voice disorders. PMID- 7471735 TI - Therapeutic plasmapheresis and cytapheresis. A review. PMID- 7471734 TI - Hyaline membrane disease in twin B following prolonged rupture of membranes for twin A. PMID- 7471736 TI - The medical malpractice closed claims study. Conducted by the National Association of Insurance Commissioners. PMID- 7471737 TI - Radioimmunoassay of plasma ethinylestradiol in the presence of circulating norethindrone. AB - A reliable radioimmunoassay (RIA) for the measurement of ethinylestradiol (EE2) in plasma after the separation of norethindrone (NET) by column chromatography has been developed. The procedure involves diethyl ether extraction, followed by celite column partition chromatography to separate NET from EE2. An antiserum against ethinylestradiol-7-(3-thiopropionic acid)-bovine serum albumin conjugate was employed. Tritiated EE2 was used as the radioligand. The separation of bound from free 3H-EE2 was carried out with dextran-coated charcoal. This RIA procedure was used to measure plasma EE2 at specified time intervals in three women following oral administration of a tablet containing EE2 (0.12 mg) and NET (1.0 mg). The results show that EE2 is rapidly absorbed after oral administration. Peak concentrations of plasma EE2 were reached within 1.0 to 1.5 hours and then declined biphasically. The EE2 alpha phase half-life values in three subjects were 1.5, 2.8 and 3.8 hours. By comparison, the half-life values for the beta phase for the same three women were 6.2, 12.3 and 14.5 hours, respectively. Plasma samples were also analyzed for NET by RIA. The half-life values for the beta phase for EE2 and NET in the three subjects were similar. PMID- 7471738 TI - A comparison of two stable prostaglandin E analogues for termination of early pregnancy and for cervical dilatation. AB - Termination of pregnancy with prostaglandin E analogues is in general associated with a lower frequency of gastrointestinal side effects than if corresponding F analogues are used. Their clinical use has, however, been limited by stability problems. In the present study the efficacy of different dose schedules of two new stable E analogues for termination of early pregnancy and for preoperative dilatation of the cervical canal was evaluated in 389 women. In early pregnant patients, vaginal administration of 75 mg of 9-deoxo-16, 16-dimethyl-9-methylene PGE2 repeated after six hours or three intramuscular injections of 0.5 mg 16 phenoxy-omega-17, 18, 19, 20-tetranor PGE2 methyl sulfonylamide administered in three-hour intervals resulted in a complete abortion in 94 to 100 per cent of the patients. Both treatments were associated with a low frequency of side effects. The 9-methylene analogue had the advantage of causing less uterine pain than 16 phenoxy-omega-17, 18, 19, 20-tetranor PGE2 methyl sulfonylamide with the dose schedules used. Single vaginal administration of 30 mg of 9-deoxo-16, 16-dimethyl 9-methylene PGE2 and one intramuscular injection of 0.5 mg of the methyl sulfonylamide analogue 12 hours prior to vacuum aspiration were equally effective in dilating the cervix in late first trimester pregnant patients. For both compounds, the frequency of side effects were lower than that previously reported for different PGF analogues administered by non-invasive routes. PMID- 7471739 TI - The interaction of phenobarbital and other anticonvulsants with oral contraceptive steroid therapy. AB - In a group of 5 women on long-term anticonvulsant and oral contraceptive therapy, the plasma ethynylestradiol (EE) concentration on 50 microgram EE daily was 11.1 +/- 4.5 pg/ml. These values were at the lower end of the range found in normal women in this laboratory taking 30 microgram EE daily (6-190 pg/ml). Four women have been studied prospectively for 3 months, over 1 cycle before and 2 cycles during phenobarbital 30 mg b.i.d. therapy. Significant falls in the plasma EE concentration were seen in two women (from 104.8 +/- 13.4 to 37.7 +/- 2.0 pg/ml and from 125.6 +/- 23.8 to 34.8 +/- 6.7 pg/ml p less than 0.01) and breakthrough bleeding was seen in both women. No changes in plasma concentrations of follicle stimulating hormone, progesterone, norethindrone or norgestrel were seen. There was a significant increase in the sex hormone binding globulin capacity from 100.7 +/- 5.8 to 133.3 +/- 1.2 nmoles/1 (p less than 0.05). These changes are consistent with the known microsomal enzyme inducing effect of phenobarbital. PMID- 7471740 TI - Inhibition of glycolysis in boar sperm by alpha-chlorohydrin. AB - The effect of alpha-chlorohydrin (3-chloropropan-1,2-diol) on the metabolism of D [U-14C]-fructose and L-[U-14C]-lactate by washed boar sperm has been investigated. Whereas alpha-chlorohydrin at concentrations as low as 0.1 mM inhibited the metabolism of fructose and led to an increase in the utilization of endogenous lactate, amounts up to 500 mM had no effect on the oxidation of added lactate. Low levels of alpha-chlorohydrin increased the cellular concentrations of fructose-1,6-bisphosphate and the triosephosphates, an effect consistent with the inhibition of glyceraldehyde-3-phosphate dehydrogenase. Although [U-14C] glycerol was rapidly oxidized by boar sperm, [U-14C]-alpha-chlorohydrin was not metabolized to 14CO2, showing that it is not converted to glycerol. Extracts obtained by sonication of boar sperm after incubation with [3-36C1]-alpha chlorohydrin, did not contain [3-36C1]-alpha-chlorohydrin-1-phosphate. This is contrary to the postulate that this phosphorylated compound is the inhibitory metabolite of alpha-chlorohydrin. PMID- 7471741 TI - Oxytocin augmentation of second-trimester abortion: safe or hazardous? AB - The role of oxytocin in augmenting the uterine activity of second-trimester instillation abortions is controversial. To evaluate the efficacy and safety of oxytocin augmentation, we studied 10,013 women who underwent hypertonic saline instillation procedures and 1,241 women who underwent prostaglandin F2a (PGF2a) instillation procedures in the United States in the period 1971 through 1975. Data were obtained through the Joint Program for the Study of Abortion under the auspices of the Center for Disease Control, a multi-center, prospective, cohort study. We found that concurrent administration of oxytocin was associated with a significantly shorter instillation-to-abortion time for hypertonic saline, but with a significantly longer time for PGF2a. For hypertonic saline, use of concurrent oxytocin did not affect either the success rate or the major complication rate; however, it did not affect the specific type of complication that occurred. For PGF2a, use of concurrent oxytocin was associated with a lower success rate and a higher major complication rate than when PGF2a was used alone; however, the relatively small number of observations and limitations in our study design may have affected our results. When we compared the 2 main abortifacients, even when we controlled for concurrent oxytocin augmentation, unaugmented PGF2a had higher major complication rates than hypertonic saline. PMID- 7471742 TI - Effect of Lippes loop on sperm recovery from human fallopian tubes. AB - Recovery of human sperm from the fallopian tubes after natural insemination have been studied in sixty women; out of these, thirty cases were fitted with the Lippes loop, type C, for at least six months. The mean sperm count/high power field (HPF X 400) and percentage of motile sperm in the cervical mucus in both control and Lippes loop groups showed no significant differences. No sperm were recovered from the fallopian tubes of the Lippes loop group, while fourteen control cases had a positive recovery of sperm from the tubal washings. The mean number of white blood corpuscles in the tubal washings of Lippes loop-fitted cases was found to be significantly higher than in control cases. The phagocytic capacity of these cells may explain the low incidence of sperm recovered from the fallopian tubes in such cases. PMID- 7471743 TI - Low dose injectable contraceptive norethisterone enanthate 20mg monthly - II. Metabolic side effects. AB - Metabolic effects of a long-acting low dose injectable contraceptive, norethisterone enanthate 20-mg, monthly injections (Neten-20), was tested in 13 women belonging to the low income groups over a period of 1 year. No change was observed in hemoglobin, hematocrit, glucose tolerance, plasma lipids, iron, calcium, or serum glutamate-oxaloacetate transaminase after treatment. Marginal rise in albumin and fall in some globulin fractions was observed. The slight fall seen in serum alkaline phosphatase could be attributed to a change in lactation status. Vitamin A, pyridoxine and riboflavin status were not altered. A peculiar aberration in the tryptophan-niacin pathway as indicated by rise in kynurenic acid excretion after tryptophan load was observed. This could be corrected by multivitamin therapy. These data suggest that the use of Neten-20 for one year does not lead to adverse metabolic effects analogous to those seen with combination type oral contraceptives. PMID- 7471744 TI - The effects of sterilisation: a comparison of sterilised women with the wives of vasectomised men. AB - In a follow-up study, women sterilised by tubal diathermy were compared with a matched group of wives of vasectomised men. Semi-structured interviews were given to a random sample drawn from a representative population. The couples were young with small families and did not have a high proportion of unplanned pregnancies or terminations. They had previously used contraception, mainly the pill or sheath. Most couples were entirely satisfied with the operation. Both groups showed an increase in pre-menstrual symptoms but there was only slight evidence that menstrual loss was affected by female sterilisation. The vasectomy couples had a higher frequency of sexual intercourse, few sexual problems and tended to have more satisfactory marriages. They had had more discussion of their decision to have the operation and the implications of counselling are considered. PMID- 7471745 TI - The effect of daily d-norgestrel (30 micrograms) on serum progesterone in Thai women. AB - Serum progesterone levels were measured by RIA in 23 Thai women during the first 3 of a 6-month treatment period of taking 30 micrograms d-Norgestrel daily and again for a 3-month period after discontinuing its use in order to determine the frequency of ovulation. During the period of use, 11 out of the 23 subjects showed no evidence of ovulation while only 1 subject failed to ovulate during the post-treatment stage. The median time for the post-treatment resumption of ovulation was 20 days with the earliest occurring before day 10 and the latest around day 85. The frequency of ovulation in women who did ovulate during the treatment period appeared to be depressed with an average of 1.8 ovulations/subject as compared to 2.4 during the post-treatment. Menstrual bleeding patterns were not significantly altered. PMID- 7471747 TI - The rabbit as an animal model to study pharmacokinetics of levonorgestrel in women. AB - In an attempt to develop an animal model to facilitate investigations on the effects of various nutritional and environmental factors on the metabolism of the contraceptive steroid levonorgestrel, the pharmacokinetics of levonorgestrel were investigated in rabbits. Six adult female rabbits received an intravenous bolus dose of 3H-levonorgestrel, 8-20 microCi, with 15-20 micrograms of carrier cold levonorgestrel per kg body weight. Blood samples were drawn at different intervals of time after the dose and the plasma drug concentration was determined both by radioisotopic dilution method and by radioimmunoassay. Plasma concentrations of levonorgestrel as assayed by radioimmunoassay agreed very closely with those obtained by isotope counting. The decline in plasma radioactivity attributable to unchanged levonorgestrel followed a triexponential pattern. Plasma radioactivity data were therefore analysed according to a three compartment open model by graphical and regression analysis. The half-lives of the three phases pi, alpha and beta representing the shallow, deep peripheral and the terminal elimination compartment, respectively, were in close agreement with those reported in women. The metabolic clearance rate (MCR) and volume of distribution (VB) of levonorgestrel expressed per unit of body weight were comparable to values reported in women. The overall observed trend of pharmacokinetics of levonorgestrel in rabbits resemble closely those seen in women. It would therefore appear that the rabbit is a suitable animal model for studying the effects of nutritional factors on the pharmacokinetics of the drug. PMID- 7471746 TI - Contraception with subcutaneous capsules containing ST-1435. Pituitary and ovarian function and plasma levels of ST-1435. AB - An RIA for a new progestin, ST-1435, was developed. Plasma concentrations of this steroid were measured in three women having three subcutaneous capsules, each containing 40 mg of ST-1435. Pituitary and ovarian functions were assessed by measuring plasma levels of LH, FSH, estradiol and progesterone. Plasma profiles of ST-1435 were characterized by high and irregular peaks all the way through the study period of 7-9 months. Concentrations below 100 pg/ml were not observed, whereas the highest peaks reached concentrations of 10.1 ng/ml. Plasma estradiol concentrations remained uniformly suppressed below 100 pg/ml, thus indicating a suppression in follicle development. As judged by constantly suppressed progesterone concentrations, no ovulations occurred during the treatment. Plasma FSH concentrations remained at the upper limit of the follicular phase, and those of LH at the lower limit of the luteal phase of the normal menstrual cycle. No midcycle gonadotropin peaks were observed. PMID- 7471748 TI - The fatty acid pattern of serum triglycerides and FFA in patients with essential hypertension of different stages, athletes, and normal subjects. AB - The fatty acid pattern of serum triglycerides and FFA in normal untrained subjects, normotensive athletes, patients with labile and stable essential hypertension as well as in hypertensives with overweight and mild hypertriglyceridemia has been evaluated by gas liquid chromatography. The most striking differences revealed the linoleic acid in triglycerides being increased in athletes and in patients with labile hypertension in comparison with normotensive untrained controls and patients with stable hypertension. On the basis of these data an enhanced need of polyunsaturated fatty acids corresponding to probands with high physical activity has been assumed in patients with an early stage of essential hypertension. The differences of arachidonic acid were not so distinct. The results can be relevant with regard to the pathogenetic role of prostaglandin precursors in the development and course of essential hypertension and atherosclerosis. PMID- 7471749 TI - Automatized determination of left ventricular systolic intervals and assessment of their significance in the epidemiology of ischaemic heart disease. AB - Knowledge of the functional state of the myocardium in patients with coronary heart disease is of essential importance for the prognosis and therapy of the disease. The PEP/LVET index is closely correlated with the ejection fraction and is a reliable criterion for separating patients with normal and disturbed function of the left ventricle, but it does not suffice for uncovering an initial haemodynamic alteration either at rest or during isometric exercise test. The good reproducibility of systolic intervals makes possible a long-term follow-up of patients, especially in cases in which direct examination methods have limited applicability. Computer processing of polygraph tracings makes possible to analyse data of large groups of patients. The accuracy of systolic intervals determination matches that of the classical reading by two independent observes. The use of computing techniques in the analysis of curves and in their statistical evaluation contributes to the standardization of the whole measuring procedure and makes possible a wide clinical use of non-invasive polygraphic methods. PMID- 7471750 TI - A survey of the opinions and attitudes of general practitioners in Kaunas to the prevention of ischaemic heart disease. AB - Fifty-four general practitioners were sent questionnaires inquiring into their attitudes to various aspects of preventive medicine and to primary prevention of ischaemic heart disease (IHD). Besides, their opinions were questioned about three risk factors of IHD (arterial hypertension, hypercholesterolaemia, impaired glucose tolerance), and these factors' values above which drug therapy should be started. It was found that the physicians tended to start drug therapy already at fairly low values of the three risk factors. A survey repeated after extensive preventive study among the population revealed no marked changes in the opinions of the practitioners questioned. PMID- 7471751 TI - Action potential characteristics of rabbit papillary muscle. AB - The ionic components of the action potential (AP) in rabbit right papillary muscle (RRPM) were determined by simple methods such as changes in external concentrations of sodium and calcium ions and addition of MnCl2 ot the perfusion fluid. Two components (fast and slow) were clearly visible in the upstroke of AP in normal Tyrode solution. MnCl2 (5 mmol/l) eliminated the slow component which forms the overshoot (OS) leaving the fast one unaffected, indicating that the former was related to a slow inward current. Mn also drastically reduced AP duration without changing the slope of phase 3 of repolarization. Reduction of Na concentration to 90 mmol/l did not affect the peak voltage of the OS but shortened AP. This low-Na solution reduced both (Vmax)f and the amplitude of the fast component. Ca-free solution reduced OS and increased AP duration. In high-Ca Tyrode (7.2 mmol/l) OS was increased and AP shortened. The effects of MN were reversed in this solution. The results suggest that the upstroke of the RRPM action potential is determined by two different ionic currents: a fast Na current responsible for the fast component and a slow inward current carried mainly by Ca, this latter being responsible for the slow component in the upstroke and for the plateau phase. The existence of propagated, Ca-dependent slow responses was demonstrated. Emphasis is placed on the fact that the major determination of the repolarization process in this type of AP is probably the inactivation of the slow inward Ca current. PMID- 7471752 TI - Effects of moderate hypoxia on action potential and contraction of rabbit ventricular muscle. AB - Experiments were performed on rabbit right ventricular strips in order to investigate the effects of moderate hypoxia on electrical and mechanical activities of this preparation. The results indicate that a slow inward current (Isi) is responsible for overshoot (OS) and the plateau phase of the action potential (AP). Hypoxia (pO2 = 65-70 mmHg; 8.66-10.0 kPa) induced a rapid shortening of AP and an increase in the phase-3 slope. It also markedly reduced twitch tension and time to peak tension, and increased resting tension. The resting potential (RP), OS and maximum rate of depolarization of phase O were not affected during 60 min of hypoxic perfusion. Isoproterenol under this condition had a positive inotropic effect and also increased AP duration (APD) measured at 60 mV, having a less pronounced effect on the plateau phase. MnCl2 in hypoxic conditions drastically reduced OS indicating that even in this condition, Isi was present. These results suggest that an increase in gK is responsible for the hypoxia-induced reduction in APD. This also partially explains the maintenance of RP during hypoxic incubation. PMID- 7471753 TI - Pulmonary vascular responses to inhalation anaesthesia in isolated dog lungs. AB - To study the mechanisms of action of four inhalation anaesthetics (diethyl ether, halothane, methoxyflurane, and nitrogen monoxide) upon the pulmonary circulation, the authors carried out 45 experiments in isolated, perfused and ventilated canine lungs. The effects of the anaesthetics were studied at 1) normotonic perfusion, 2) enhanced pulmonary blood flow, 3) microembolism-induced pulmonary hypertension. In the first two-experimental series, no effects of the test anaesthetics on the pulmonary vascular responses became manifest; at microembolism-induced pulmonary hypertension, halothane lowered the pulmonary vascular resistance, whereas diethyl ether stabilized the elevated vascular tone. Methoxyflurane and nitrogen monoxide had no marked effects on the pulmonary vascular responses. On the basis of their experiences and of data published in the literature the authors conclude that there exist regional mechanisms of action of anaesthetics on the lung vessels, activated by the release or action of mediators. PMID- 7471754 TI - Relationship between smoking and ischaemic heart disease mortality in male population of Kaunas, aged 45-59 years. AB - Out of 2392 men aged 45--59 years, examined in accordance with the WHO standard methods for detection of risk factors of ischaemic heart disease (IHD), within the follow-up period from March, 1972, till February, 1978, 96 persons died. Considerable differences were found in the proportion of ex-smokers and non smokers in the group of individuals without IHD and among patients with IHD. Among all deceased subjects in those deceased of IHD and among the persons without IHD the proportion of smokers was considerably higher than the proportion of non-smokers. In the smokers the most frequent cause of death were malignant tumours. Overall mortality and mortality due to IHD were markedly higher in smokers as compared to ex-smokers. Considerably lower values of systolic, diastolic arterial pressures and cholesterol, lower body mass, and lesser skin fold thickness were found in the living smokers. Blood sugar levels were not different in smokers, ex-smokers, and non-smokers. In the light of the present study, smoking is the factor most increasing the risk of death of malignant tumours and considerably increasing the probability of death of IHD. PMID- 7471755 TI - Education and the risk factors of ischaemic heart disease. AB - Screening for the risk factors of ischaemic heart disease among the male residents of the district of Prague 4 aged 40--49 years was undertaken under the "National multifactorial primary preventive study of myocardial infarction and stroke". Of the 11 091 men invited, 5395 met the defined criteria and were admitted to the study. 58% of them were included in the risk group. The level of attained school education was ascertained at the initial examination. The lowest percentage of persons with hypercholesterolaemia, overweight, elevated blood pressure and smokers was found among university graduates. The results are compared with data of other studies and possible causes of the findings are discussed. PMID- 7471756 TI - Prognosis of pulmonary hypertension in chronic obstructive pulmonary disease. AB - Studies performed since 1968 have shown that in patients with chronic obstructive pulmonary disease (COPD) the prognosis is worse in cases with pulmonary artery hypertension (PAH), especially when the mean pulmonary artery pressure (PPA) is greater than 30 mmHg. Author's data are based on the study of a group of 175 patients with COPD, catheterized between 1968 and 1972. The fate of the patients, at least five years after the initial catheterization, was known in all the cases. The survival rate was calculated according to the "actuarial method". The results suggest that PPA has a good prognostic value, but that FEV1 and PaCO2 are as good predictors of mortality as PPA. Furthermore the influence of age is to be taken into account: prognosis is worse in patients older than 65 years. Longitudinal studies of pulmonary haemodynamics was made in 85 patients who underwent two right heart catheterizations with an interval of greater than 3 years (mean = 5 years) between them. The results indicate that PPA varies very slightly: the average increase for the whole group is 3.1 mmHg, that is less than 1 mmHg per year. There was a good negative correlation between chronological variations of PaO2 and PPA. A long survival time could be observed after the first heart failure episode or after ascertaining pulmonary hypertension. PMID- 7471757 TI - Doppler echocardiography in diagnosis of mitral insufficiency. AB - By means of pulsed Doppler echocardiography (PDE) the authors investigated 53 patients who subsequently underwent heart catheterization and left ventriculography. The results of PDE examination agreed with ventriculography in 29 out of 31 patients with mitral regurgitation (PDE sensitivity = 94%). In 22 patients with no angiographic evidence of mitral regurgitation PDE exhibited two false positive results (PDE specificity = 91%). A semi-quantitative assessment of regurgitation by predetermined PDE criteria was successful in differentiating a group of severe mitral insufficiency (MI) from less significant MI (= mild and moderate MI angiographically). The agreement with ventriculographic classification of MI was 87 and 86%, respectively. The presented results clearly indicate that PDE, as a non-invasive methods, is precise enough in detecting mitral regurgitation and can be recommended for application in adult cardiology. PMID- 7471758 TI - Echocardiography in infective endocarditis. AB - Two cases of infective endocarditis are reported. In both, vegetations on the cardiac valves characteristic of endocarditis were documented by echocardiography and confirmed at surgery in one of them. The various features of vegetative endocarditis on the echocardiogram are described. Differentiation of these echoes from those produced by other morbid states is discussed. Echocardiography is considered a useful non-invasive technique in the diagnosis of infective endocarditis. Cardiac surgery is usually found to be necessary in addition to medical therapy, when echoes characteristic of vegetative endocarditis are recorded by echocardiography. PMID- 7471759 TI - Ultrastructural investigation of renomedullary interstitial cells in "endocrine kidney". AB - In the renomedullary interstitial cells of rats with Selye's "endocrine kidney" in the phase of beginning hypertension there appeared bilateral stress hypergranulation and an ultrastructural pattern of intensive prostaglandin synthesis. In the interstitial tissue of the left medulla there appeared light and dense (fibroblast- or macrophage-like) cells. Along with the stabilization of hypertension a relative degranulation of interstitial cells in the right kidney and an absolute degranulation in the left kidney took place. Necrobiotic changes, appearing in the ischaemic kidney, reflected an exhaustion of morphological substrates of prostaglandin synthesis. The changes in the interstitial cells, possessing endocrine activity, present the decisive links in the genesis of experimental malignant hypertension in rats. PMID- 7471760 TI - The handedness of rhesus monkeys. III. Consistency within and across activities. AB - The results of this study of the hand preferences of rhesus monkeys on three different tasks are threefold: (1) When retested on the same task at intervals exceeding one month virtually all individuals prefer the same hand as they did during the original test, (2) When retested on the same task, the strength of hand preference displayed by each individual is increased. (3) When tested on differing tasks, monkeys display little consistency in the laterality of hand preference or the strength of handedness expressed during different tasks. Many authors have concluded that the lack of obvious intertask consistency in the laterality of hand preference expressed by lower primates constitutes evidence for a corresponding lack of consistent laterality in the cerebral control of this behavior. This has led to them to conclude that cerebral dominance probably does not exist in these animals (Deuel, 1975; Warren, 1977). However, where data is available from the literature, including the present study, all reports show monkeys to more frequently prefer the same hand on all of the unimanual tasks they were given than would be expected by chance alone. This finding suggests that there is a weak tendency for consistent lateralization of hand usage in the monkey. Presumably, there is a corresponding predominance of the cerebral hemisphere contralateral to the preferred hand over its mate. Other studies consistent with the concept of cerebral predominance in the monkey were reviewed. These findings do not constitute evidence for cerebral dominance in the monkey akin to that found in man. They do suggest that when performing certain activities, monkeys may have one hemisphere predominant over the other even though the degree and laterality of predominance may vary greatly from one individual and task to another. PMID- 7471761 TI - The hemispheric specialization for linguistic and non-linguistic tactile stimuli in third grade children. AB - Thirty third-grade children were given two dichaptically presented tests of hemispheric specialization. Specialization for linguistic stimuli was measured by a letters task and specialization for spatial stimulus was measured by a nonsense shapes task. The results showed a significant right hemisphere processing advantage for tactually presented spatial stimuli. There was no significant processing advantage for either hemisphere with tactually presented linguistic stimuli. The right hemisphere specialization for tactually presented spatial stimuli was present for both boys and girls. Thus there was no evidence of sexual bimorphism in the neurological organization of third grade children for spatial processing. PMID- 7471762 TI - Absence of any developmental trend in right hemisphere superiority for face recognition. AB - Upright or inverted pictures of faces were presented bilaterally, with one face falling in each visual hemifield, to right-handed children aged seven, ten and thirteen years. When a small set of four stimulus faces was used with each child (Experiment I) both boys and girls were found to be better at recognising upright faces falling in the left visual hemifield (LVF). For inverted faces there was no difference between the visual hemifields. When a set of forty stimulus faces was used (Experiment II), boys only were found to be better at recognising upright faces falling in the LVF. These results are interpreted in terms of superiority of the right cerebral hemisphere for processing upright faces. No developmental differences in the degree of this superiority were found in the age range studied. PMID- 7471763 TI - Lateral eye movement and handedness as measures of functional brain asymmetry in learning disability. AB - Evidence is presented which suggests that learning disability is related to lateral asymmetry. Significant differences were found in lateral eye movement and handedness between a learning disabled group and a control group of normal learners. Left lateral eye movement and right handedness was the predominant pattern exhibited in learning disability. These results support Day's notion that left lateral eye movement is associated with educational difficulties. They also provide evidence concerning Orton's hypothesis that mixed cerebral dominance characterizes the learning disabled. PMID- 7471764 TI - Retrieval time in a monaural learning task. AB - This study examined the possible role of retrieval and/or storage processes in auditory asymmetry phenomena. Subjects pressed a left- or right-hand key labeled "yes" or "no" to indicate whether a monaural tone had just been presented to the ear specified by a left or right probe light. Subjects were faster in indicating that the probed ear had been stimulated than in indicating that it had not been stimulated. Subjects also responded faster when the location of the response key and probe light corresponded than when they did not. The methodological implications of the latter finding for research on hemispheric dominance were discussed. Finally, there was no evidence that retrieval time varied as a function of either the ear probed or the ear stimulated. PMID- 7471765 TI - Subject variables and cerebral organization for language. AB - The present study investigated the usefulness of a variety of subject variables that have been proposed as having predictive value for determining cerebral organization for language. To accomplish this, a total of 373 subjects (117 left handers and 256 right-handers) were given 240 trials of a consonant-vowel dichotic listening task to assess direction and degree of language lateralization. Each subject was also classified on the basis of eight subject variables (handedness, strength of handedness, familial sinistrality, writing hand posture, sex, sighting dominance, preferred footedness, and overall laterality). The results of the study indicated that left hemisphere language processing is very pervasive and that most of the subject variables examined were not very useful predictors of language lateralization. In addition, surprisingly, footedness and not handedness was the single best predictor of cerebral organization for language. PMID- 7471766 TI - Guaranteed stimulus-processing in the evaluation of memory and learning. AB - A test of memory and learning with guaranteed initial stimulus-processing was validated and is here presented with norms for 70-79 year old and 80-89 year old community residents vs. nursing-home residents. The test procedure also allows incidental observation of stereognosis, object-naming, and left-right orientation, screens for depression, and elicits rapid retrieval of words from semantic categories--all common aspects of evaluation for dementia. A validation study showed a significant difference between 21 moderately impaired and 21 unimpaired elderly nursing home residents grouped by their scores on Blessed, Tomlinson and Roth's (1968) "mental test". Reliability was determined by coefficient alpha, which yielded an estimated correlation of .84 between recall (retrieval) scores and errorless true scores. Storage scores were correlated .72 with retention after three weeks (p. less than .01), and storage and retrieval were not significantly correlated with each other when mental status was partialed out. Storage impairment appeared more significant in dementia than retrieval impairment, which occurred in normal aging as well. Norms for community active vs. nursing-home-residing 70 and 80 year olds are provided which allow the separate evaluation of total recall, storage, retrieval, and response to reminders. PMID- 7471767 TI - Ideatory apraxia in a left-handed patient with right-sided brain lesion. AB - A case is described of a left-handed patient with a circumscribed right-sided posterior brain lesion, who presented with a neuro-psychological syndrome of Wernicke's aphasia, ideomotor and ideatory apraxia. The aphasia and ideomotor apraxia cleared within 10 days, while ideatory apraxia persisted. Ideatory apraxia therefore was not dependent on the language disorder, nor was it related to ideomotor apraxia. On the basis of various neuropsychological examinations, the nature of the apraxic movement disorder in this case is discussed. To our knowledge, this is the first case of ideatory apraxia with right-sided brain damage described in the literature. A particular feature of this patient is that obviously language and the motor functions underlying performance in tests for ideomotor apraxia had a bilateral hemispheric representation whereas a unilateral lesion was sufficient to bring about persistent ideatory apraxia. PMID- 7471768 TI - Lateralized asymmetries in self-perception. AB - Thirty right-handed males and females, displaying happy and sad expressions, compared left-left and right-right composites of his/her own face. For each pair of composites, depicting one emotional state, subjects chose that composite which (a) looked most like themselves and (b) depicted the more intense emotion. The results for females failed to support the contention that the right hemisphere specializes in the perception of negative affect whereas the left hemisphere is prominent for positive states. Finally, asymmetries were more pronounced in females rather than males. PMID- 7471769 TI - The hemispheric side of neocortical damage does not affect memory for unidimensional position. An experiment wih Posner and Konick's test. AB - The role of neocortical areas in visuo-spatial memory was investigated by testing unilateral hemisphere-damaged patients for their ability to reproduce the position of a dot on a horizontal line after a delay. One hundred and eleven patients, divided into 4 groups on the basis of the side of damage and of the presence or absence of visual field defects, and 56 controls were tested. There were 2 sets of experimental conditions: (i) a free delay condition of 0, 20 or 180 sec., and (ii) a 20 sec. delay condition filled with 2 kinds of interpolated perceptual activity. Forgetting was significantly affected by presence of hemisphere damage, but not by the side and the intrahemispheric locus of lesion. It was concluded that the memory ability involved by this task is not related to the functioning of a specific cortical area. PMID- 7471770 TI - Selecting retrieval cues for early-childhood amnesia: implications for the study of shrinking retrograde amnesia. AB - A method for selecting semantic cues for early childhood memories is presented. The written records of episodes submitted by the subjects in the study of Crovitz and Harvey (1979) were coded into three-word strings of words in accordance with the gist of the memories. Nouns which described many retrievals (Set A) and nouns which had described few retrievals (Set B) were presented to an independent group of young adults as prompts for early childhood episodes. Prompting with semantic cues gave a fast rate of retrieval, with nouns in Set A cuing early childhood memories more successfully than nouns in Set B. The difference between words in Set A and Set B was found not to be a function of the frequency of usage of these words in the language. Implications for selecting cues to prompt retrieval of memories during the period of shrinking retrograde amnesia are discussed. PMID- 7471771 TI - Measuring familial sinistrality. PMID- 7471772 TI - Nonaphasic misnaming and other neurobehavioral features of an unusual toxic encephalopathy: case study. AB - A patient with propranolol-induced mental status changes was studied during and after the period of propranolol intoxication. While intoxicated he manifested the syndrome of "non-aphasic misnaming," did poorly on the nonverbal portions of the WAIS, and exhibited perseveration and variability of performance. Our observations clarify the nature of the neurobehavioral disturbances in toxic metabolic encephalopathies. PMID- 7471773 TI - Comments on the functional asymmetries of the lower extremities. AB - Several tasks have been used to establish laterality in the lower extremities, but most of them are not satisfactory. The lower extremities are involved in various posturomotor, locomotor and operant activities, and several kinds of differentiated bilateralizations of the lower limbs can be identified. A distinction between leggedness and footedness seems warranted. The complexity of the functional asymmetries of the lower extremities requires new methods in both research and clinical testing, with elecctrophysiological procedures as a promising approach. Available evidence indicates that the functional asymmetries of the axial neuromotor mechanisms have to be considered in the study and the assessment of those of the lower extremities. PMID- 7471774 TI - Amusia due to rhythm agnosia in a musician with left hemisphere damage: a non auditory supramodal defect. AB - A case of a severe receptive and expressive amusia in a professional musician following a left hemisphere vascular stroke is reported. Recognition and production of single tones and random tone sequences were found to be surprisingly well preserved. In contrast, the recognition and production of simple rhythm patterns were grossly disturbed. It is suggested that amusia is due to the demonstrated rhythm disturbance. Moreover, it has been found that the defect in recognition and reproduction of rhythms was manifested regardless of the modality of perception, i.e. whether rhythm patterns were perceived by audition, vision, or touch. Therefore, the disturbance of rhythm abilities is supramodal in nature, based probably on the perception of time microintervals. Thus, this type of amusia is a result of a non-auditory supramodal defect, i.e., an impairment of temporal pattern recognition. PMID- 7471775 TI - Sex differences in adults' spatial and verbal memory span. AB - Spatial span (Corsi's block-tapping test) and verbal span (Wechsler Digits Forward test) were measured in 300 medical students (150 males and 150 females). Significant differences pointing to a better performance of males were found on both spatial span (p less than 0.001) and verbal span (p less than 0.05). PMID- 7471776 TI - The role and limitation of CT in craniocerebral trauma. AB - CT is being utilized more often in evaluation of craniofacial and craniovertebral trauma. It is far superior to conventional pluri-directional tomography, since not only the complex fracture but also communication with the orbital or cranial contents can be demonstrated. The extend of soft-tissue trauma can also be better evaluated. However, there are limitations in utilizing CT as the sole modality in evaluating craniocerebral trauma. This article deals with the limitations of utilizing CT alone in evaluation of trauma. PMID- 7471777 TI - CT in trauma of the base of the skull and its complications. AB - Diagnosis and accurate delineation of fractures involving the base of the skull can easily be done with newer generation computed tomography (CT) scanner. CT also demonstrates associated complications, such as hemorrhage, brain injury, and leakage of cerebrospinal fluid (CSF), not seen in plain skull radiography or tomography. PMID- 7471778 TI - Computed tomography in child abuse head trauma. AB - Forty-five head trauma cases from 177 child-abuse patients are reported. Intracranial injuries include acute epidural and subdural hematomas, subdural effusion, cerebral contusion, and atrophy (subdural effusion and cerebral contusion being most frequently seen). Incidence of skull fracture among the 45 patients is about 38%. Sixty-five percent of the 17 skull fracture patients also had intracranial injury, and 78% of the 28 patients with normal skulls had intracranial injury. CT should be the primary tool for evaluating child-abuse head trauma. PMID- 7471779 TI - Delayed sequelae of head injury. AB - The evaluation of delayed sequelae of head injury is greatly facilitated by CT. This non-invasive technique provides us with the opportunity to evaluate the head trauma patients sequentially to detect the development and clinical significance of post-traumatic sequelae like delayed intracerebral hemorrhage, hydrocephalus, and post-traumatic atrophy. PMID- 7471780 TI - The role of computed tomography in acute head injury. AB - Computed tomography (CT) of the head provides the ability to rapidly evaluate patients with acute head injuries. Moreover, CT aids in surgical planning, prognosticating outcome and recovery time, and monitoring nonsurgical injuries. The authors review the ways in which CT is employed in a multiple trauma center. PMID- 7471781 TI - Ct in spinal trauma. AB - The radiological evaluation of patients with acute spinal trauma has always been a challenging problem. Multiple radiological procedures are often necessary for complete evaluation of the extent of spinal injury. CT provides an ideal modality whereby accurate assessment of displacement of bony fragments as well as associated spinal cord and nerve root injury can easily be performed, eliminating the need for difficult radiological procedures. PMID- 7471782 TI - Computed tomography of hepatic fracture: a case report. AB - A case of hepatic fracture is presented in which computed tomography (CT) played a major role in the diagnosis. The fracture, which was suggested on scintigraphy, was easily demonstrated on CT as a low-density hematoma. The advantages and potential use of CT in this clinical setting are discussed. PMID- 7471783 TI - Demonstration of unsuspected malignant disease of the pericardium by computed tomography. AB - Massive pericardial effusion associated with mediastinal neoplasm was first diagnosed by findings at computed tomography (CT) in two patients. In each case the CT findings prompted urgent pericardiocentesis for relief of pericardial tamponade. One of the patients was subsequently found to have a widely disseminated renal papillary adenocarcinoma, and the other had fibrosarcomatous mesothelioma of the pericardium. The role of CT in the diagnosis of pericardial disease is discussed. PMID- 7471784 TI - Computed tomography (CT) scanner in Peking (Beijing), China. PMID- 7471785 TI - The relationship between cell injury and osmotic volume reduction: II. Red cell lysis correlates with cell volume rather than intracellular salt concentration. PMID- 7471786 TI - Control of canine kidney cortex slice volume and ion distribution at hypothermia by impermeable anions. PMID- 7471787 TI - Metabolic changes in dog kidney during cryopreservation. PMID- 7471789 TI - Experimental cryosurgery on bone: a light and electron microscopical investigation. PMID- 7471788 TI - The effect of organ preservation on polysomal messenger RNA. PMID- 7471790 TI - Ultrastructural and thermocouple evaluation of rapid freezing technique. PMID- 7471791 TI - A study of the final period of freeze-drying and determination of residual moisture of dry biological materials. PMID- 7471792 TI - Survival and recovery of cryopreserved rat platelets. PMID- 7471793 TI - Permeability of the 17-day fetal rat pancreas to glycerol and dimethylsulfoxide. PMID- 7471794 TI - Survival of frozen-thawed fetal rat pancreases as a function of the permeation of dimethylsulfoxide and glycerol, warming rate, and fetal age. PMID- 7471795 TI - An optimum method for the introduction or removal of permeable cryoprotectants: isolated cells. PMID- 7471796 TI - The vascular effect of cold injury. PMID- 7471798 TI - Abstracts of papers presented at the international symposium survival in cold (hibernation and other adaptations) July 2-5, 1980, Prague, Czechoslovakia. PMID- 7471799 TI - Atrial fibrillation. PMID- 7471797 TI - Lipid participation in intracellular freezing avoidance mechanisms of lettuce seed. PMID- 7471801 TI - Neural development Part II. Neural Development in model systems. PMID- 7471800 TI - Obesity and the heart. PMID- 7471802 TI - Low concentration halcinonide cream in the topical management of atopic dermatitis in pediatric patients. AB - Seventy-nine patients with atopic dermatitis were treated in a double-blind paired comparison fashion with halcinonide cream (0.025%) or its placebo control. The overall therapeutic response was excellent in thirty-seven (47%) patients treated with halcinonide cream (0.25%) as compared with twenty-two (28%) patients treated with the placebo cream. In the comparative response, halcinonide was considered superior in forty-six (58%) patients, and the placebo was considered superior in fifteen (19%). Statistically, halcinonide cream was shown to be significantly superior to the placebo cream (p less than 0.001). Side effects noted in five patients included burning, stinging, and pain, due in most cases to both halcinonide and the placebo cream. PMID- 7471803 TI - Conversations on allergy and immunology. PMID- 7471804 TI - Warts. PMID- 7471805 TI - Recurrences of herpes zoster. PMID- 7471806 TI - Nasal glioma. AB - Nasal gliomas are rare, benign, congenital neoplasms of the skin, occurring in infancy, and usually appearing over the bridge of the nose. A case of extra-nasal glioma in an eight-month-old girl is presented herein. PMID- 7471807 TI - Onycholysis of the thumbs probably due to a phototoxic reaction from doxycycline. AB - Three members of a party climbing to the 18,640 foot level of Mt. Kilimanjaro in Africa experienced onycholysis or partial detachment of the thumbnails. It is believed that the combination of daily doxycycline administration and intense sunlight exposure resulted in a phototoxic reaction. PMID- 7471808 TI - Congenital vellus hamartoma. PMID- 7471809 TI - Pulmonary manifestations of tuberous sclerosis. AB - Lung involvement in tuberous sclerosis is rare. Those patients with lung involvement differ from the usual tuberous sclerosis patient in that they are predominantly women, are of normal intelligence, and are less likely to develop epilepsy. The lung disease is often fatal, with cor pulmonale and pneumothorax as the major causes of death. A review of the radiologic and pathophysiologic findings of the lung process are also presented in the following case report. PMID- 7471810 TI - Cytogenetical studies on two terrestrial isopods (Crustacea), Philoscia lodnensis Ramakrishna and Porcellio rathkei Brandt. AB - The diploid number of chromosomes in two species of isopods namely, Philoscia lodnensis and Porcellio rathkei was found to be 16 and 50, respectively. The chromosomes have localized centromeric positions. Female heterogamety was found to exist in P. rathkei. PMID- 7471811 TI - Spectral evidence of interaction of spice constituents with hepatic microsomal cytochrome P-450. AB - Spectral evidence has been obtained of the interaction of constituents of two commonly used spices, capsicum and cinnamon, with hepatic microsomal cytochrome P 450. It is proposed that influences of spice constituents on hepatic mixed function oxidase activities may merit consideration during the investigation of some health-related problems characteristic of, or peculiar to, a given region or cultural group. An example considered relates to the evaluation of certain 'traditional medicines'. PMID- 7471812 TI - On the chromosomes of two species of jassids (Cicadellidae: Homoptera). AB - Chromosome number and and male meiosis of Hishimonus phycitis and Chiasmus arori of the sub-family Deltocephalinae (Family: Cicadellidae) is discussed. The diploid chromosome number in H. phycitis was found to be 2n = 11-12 with a sex mechanism of the XO:XX type, while 2n = 16 with a sex-mechanism of XX:XY has been observed in the case of Chiasmus arori. The gonial chromosomes in H. phycitis are gradually seriated, while the diploid complement of C. arori is characterized by a pair of large chromosomes. The mode of male meiosis follows the typical pattern of the family Cicadellidae and is essentially similar in the two species. PMID- 7471813 TI - Chromosome elimination in Ctenocephalides orientis (Siphonaptera). AB - Somatic and egg squashes of Ctenocephalides orientis exhibited a chromosome complement of 2n = 14,XY/XX, whereas the spermatocytes showed 2n = 15,XY1 Y2. The sex trivalent was restricted to spermatogenesis; in oogenesis only one sex bivalent was formed. Elimination of one of the Y chromosomes from the somatic cells was evident. PMID- 7471814 TI - Intergeneric hybridization and speciation between two leptosporangiate ferns. AB - The oosphere of Pteridium aquilinum is surrounded by a 'special membrane' which does not appear to resist the fertile spermatozoon of Gymnogramme calomelanos var. chrysophylla and monospermy results. Our hybridization findings also indicate that the membrane maintains speciation. The spermatozoon of Gymnogramme is strongly attracted but some injury is sustained during its passage through the mucilage of the archegonium neck. Nevertheless, rupture of the membrane, herein designated the cortical complex of speciation (CCS), is followed by expulsion of the oosphere-cytoplasm for which an osmotic mechanism is proposed. PMID- 7471815 TI - Bone marrow chromosomes in Corvus species (Corvidae: Passeriformes: Aves). AB - The diploid number of 80 +/- chromosomes has been established for two species of Indian crows, viz. Corvus splendens splendens Viellot, and Corvus macrorhynchus culminatus Sykes. In addition to the usual macro- and microchromosomes, there exist certain elements in both cases, falling in between the two different size categories of chromosomes. The number of these medium-sized, and of the macrochromosomes, was found to be 6 and 14, respectively, in both species. All chromosomes included in these two categories depict the clear position of their centromeres, while in the microchromosomes the centromeric position is not distinct. The sex-chromosome mechanism which has been studied only in Corvus splendens splendens, is of the ZZ and ZW type in the male and female, respectively. PMID- 7471816 TI - Morphological abnormalities of lymphocytes from patients with advanced solid tumours. AB - Characteristic morphological abnormalities were found to be present on lymphocytes from patients with advanced solid tumours. Numerous microprojections were characteristically present on human peripheral blood lymphocytes from cancer patients while non-cancer subject control lymphocytes ('normal' lymphocytes) appeared significantly smoother. Incubation of normal lymphocytes in cancer serum resulted in increased occurrence of these microprojections whereas incubation of normal cells in normal heterologous serum resulted in no apparent surface changes. Incubation of cancer lymphocytes in levamisole, an immunostimulant, did not abrogate surface projections. PMID- 7471818 TI - Blue spruce (Picea pungens) tissue and cell culture. AB - Blue spruce (Picea pungens) cell cultures were established from leaf and stem sections of an adult tree on a range of media of which BSPM4 (B5 medium plus 10 microM 2,4-D and 1 microM kinetin), and BSPM 6 (MS medium plus 5 microM 2,4-D and 1 microM kinetin) were optimal. Cytological investigations showed a diploid karyotype of 24 metacentric or sub-metacentric chromosomes. The karyotype and ploidy level remained constant over repeated subculture at rapid growth rates. Picea pungens chromosomes were found to be excellent cytological material because of their size and ease of visualization from stem callus. The organogenic potential of the leaf and stem callus so far seems to be restricted to extensive greening, vascular differentiation and pseudo-bud development. PMID- 7471817 TI - Inhibition of urethan induced lung tumour growth in strain A mice by 5-methyl cytidine. AB - In two in vivo experiments, low doses of 5-methyl cytidine significantly suppressed the growth of urethan-induced lung tumours in strain A mice while higher doses were less effective. This inverse dose-response relationship mimics the previously reported effect of 5-methyl cytidine on mammary tumour growth in C3H mice. The finding that 5-methyl cytidine inhibits lung tumour growth as well as mammary tumour growth suggests this nucleoside may be an effective inhibitor of solid tumour growth in general. A study of the effect of 5-methyl cytidine on mouse lung tumour cell viability in vitro suggests that the direct cytotoxic effect of 5-methyl cytidine is dependent on the cell cycle. Since cell-cycle dependent agents do not inhibit lung tumour growth in vivo, 5-methyl cytidine probably inhibits mouse lung tumour growth indirectly by some effect on the host rather than by a direct cytotoxic effect on the lung tumour. PMID- 7471819 TI - An autosomal dominant inheritance for multiple sunlight-induced malignancy in a patient without abnormalities in DNA repair or replication. Repository identification No. GM2881. PMID- 7471820 TI - Studies on endoreduplication. II. Spontaneous occurrence and cellular kinetics of endoreduplication in PHA-stimulated tonsillar lymphocytes. AB - Cells with spontaneously endoreduplicated chromosomes were observed in tonsillar lymphocyte cultures from the second to the seventh day of culture, although at a low frequency. Cell cycle kinetics leading to endoreduplication were examined by the 5-bromodeoxyuridine (BrdU)-acridine orange method. The diplochromosomes showed four fluorescent patterns depending on the period during which BrdU was incorporated into the DNA. The kinetic data also revealed two populations among the cells with endoreduplicated chromosomes: one with a shorter (36 or 48 h) and the other with a longer cell cycle time. The existance of the latter population was shown by the fact that cell without differentially stained chromatids were observed many hours after BrdU addition (up to 120 h). This suggests that some cells with endoreduplicated chromosomes originate from cells arrested in G2. PMID- 7471822 TI - The location of chromosome breaks in Bloom's syndrome. AB - The lymphocytes of a patient with Bloom's syndrome were examined for chromosome abnormalities. The break points involved in the abnormalities in this patient were found to be nonrandom in their distribution, as was previously described in the literature in another case. It is preliminary proposed that, in both cases, many more breaks occur in chromosomal areas which replicate their DNA late in the S phase of the cell cycle than would be expected by chance. The particularly severe manifestations of the disease in this patient, including an unusually high percentage of cells with chromosome abnormalities, tend to confirm the suggestion that Bloom's syndrome may not simply be the result of one structural mutation. PMID- 7471821 TI - Nucleolar organizer regions and constitutive heterochromatin in polyploid species of the genus Odontophrynus (Amphibia, Anura). AB - Chromosome banding patterns obtained by ammoniacal silver staining (Ag-AS) and alkaline Giemsa (CBG) have been analysed in several amphibian species of the genus Odontophrynus from South America. Ag-AS bands were found at secondary constrictions, mainly of chromosomes 4 and 11. The CBG technique revealed centromeric and telomeric constitutive heterochromatin on almost all chromosomes of these species. Moreover, intercalary bands were found at particular sites of several chromosomes. Some inter- and intra-population polymorphisms were found for the Ag-AS and C-banding patterns. The species variability in the number and position of the Ag-AS bands, known to be regions of active ribosomal cistrons, as well as the specific sites of intercalary heterochromatin, are used to discuss the possible evolutionary relationships among these species. PMID- 7471823 TI - Cytomorphology and behavior of late bone marrow and peripheral blood erythroid cells in experimental hemolytic anemia. PMID- 7471824 TI - The cell as a membranous network under microscope. PMID- 7471825 TI - Lung punctures. PMID- 7471826 TI - Effect of chronic pressure overload on the maximal rate of pressure fall of the right ventricle. AB - The maximal rate of fall in right ventricular pressure (negative dp/dt) was evaluated in 34 patients. Eight had normal pulmonary arterial pressure. Seventeen had pulmonary arterial hypertension, and nine had pulmonary arterial hypertension with right ventricular failure. The right ventricular maximal negative dp/dt in patients with normal pulmonary arterial pressure was 170 +/- 20 mm Hg/sec. In patients with pulmonary arterial hypertension not accompanied by right ventricular failure, this value was 670 +/- 60 mm Hg/sec; and in patients with right ventricular failure, it was also 670 +/- 60 mm Hg/sec. This was higher than in control subjects (P less than 0.001). The maximal positive dp/dt was also higher in patients with pulmonary hypertension, regardless of the presence of right ventricular failure. Right ventricular maximal negative dp/dt correlated with right ventricular maximal positive dp/dt (r = 0.72). Right ventricular maximal negative dp/dt in patients who were not in right ventricular failure correlated linearly with pulmonary arterial systolic pressure (r = 0.83) and pulmonary arterial diastolic pressure (r = 0.83). At any level of pulmonary arterial systolic pressure, right ventricular maximal negative dp/dt in patients with right ventricular failure was lower than in patients with the same level of pulmonary arterial hypertension who were not in failure. These observations indicate that right ventricular maximal negative dp/dt is dependent on load. Even in the presence of right ventricular failure, right ventricular maximal negative dp/dt exceeded values in control subjects. PMID- 7471827 TI - Life-threatening cold and exercise-induced asthma potentiated by administration of propranolol. AB - We present two patients with acute respiratory failure secondary to cold and exercise-induced asthma. Neither patient had a prior history of asthma, and both had recently been placed on therapy with propranolol for treatment of hypertension. We discuss the postulated mechanism of bronchospasm in these patients. In addition, we urge aggressive searching for any change in respiratory symptoms in patients receiving propranolol. PMID- 7471828 TI - Surgical treatment for chest pain in mitral valve prolapse. AB - The case of a 40-year-old woman with mitral valve prolapse and severe atypical chest pain is presented. The diagnosis was confirmed by phonocardiographic, echocardiographic, and angiocardiographic studies. The electrocardiogram revealed an ischemic pattern of ST-T on the anterior and inferior wall. Coronary angiographic studies showed normal coronary arteries. The patient's long standing, prolonged, disabling atypical chest pain could not be relieved with medical therapy, despite the administration of beta-adrenergic blocking agents, calcium antagonists, and short-acting nitrites during a 30-month period. Thus, the prolapsed mitral valve was replaced with a Hancock xenograft. After 12 months the patient is totally free of symptoms, without any treatment and with a normal ECG. This excellent surgical result could be explained on the basis of the valvular theory of chest pain in mitral valve prolapse, suggesting that pain is promoted probably by a regional imbalance between oxygen availability and consumption, because of the excessive papillary muscular stretching produced by the prolapse. To our knowledge, this is the first published report of successful surgical treatment of chest pain in mitral valve prolapse. PMID- 7471829 TI - Tricuspid atresia with double-outlet left atrium. AB - Clinical and pathologic findings of an unusual case of cardiac malformation are presented. The main features were those of atresia of the right atrioventricular valve associated with two distinct atrioventricular orifices connecting the morphologically left atrium to the underlying morphologically left ventricle. Other distinguishing characteristics were ostium secundum atrial septal defect, normally related great arteries, with the aorta arising from the main ventricular chamber and the pulmonary artery from the anterior and right-sided outlet chamber, and infundibular and valvular pulmonary atresia. PMID- 7471830 TI - Idiopathic pulmonary hypertension associated with nodular pulmonary infiltrates and portal venous thrombosis. AB - A case of idiopathic pulmonary hypertension is associated with portal venous thrombosis. The patient's chest roentgenogram revealed numerous nodular densities which at necropsy were found to have been caused by an exuberant fibroblastic reaction around small pulmonary arterioles. This is an example of the unusual association of idiopathic pulmonary hypertension and portal venous thrombosis, and to our knowledge, the first report of roentgenographically visible pulmonary nodules secondary to the pathologic alterations of idiopathic pulmonary hypertension. PMID- 7471831 TI - Restrictive cardiomyopathy in pseudoxanthoma elasticum. AB - A unique case of severe and diffuse endocardial fibroelastosis covering the entire left ventricle and associated with pseudoxanthoma elasticum is presented. The clinical picture was that of an apparently primary restrictive cardiomyopathy. Specific histologic lesions could be recognized in the endocardium. PMID- 7471832 TI - Echocardiographic features of flail aortic porcine valve. PMID- 7471833 TI - Detection of a subvalvular vegetation in endocarditis by two-dimensional echocardiography. Therapeutic implications. PMID- 7471834 TI - Mitral stenosis and COPD. PMID- 7471835 TI - Cost containment in a hospital program for home oxygen therapy. PMID- 7471836 TI - Perforation of thoracic aortic aneurysm. A complication of mediastinoscopy. PMID- 7471837 TI - Pulmonary arterial laceration secondary to subclavian vein catheterization. PMID- 7471838 TI - Needle aspiration of the lung in complex pneumonias. AB - In 39 patients with treated unresponsive pneumonia complicated by life threatening underlying disease, direct needle aspiration of the lung was performed to establish a cause. An infectious agent was detected in 18 patients (46 percent), a nonbacterial cause was found in one, and cure of a recent infection was substantiated in six patients. An incorrect diagnosis was made in seven patients (one false-positive and six false-negative), and an indeterminant answer was obtained by needle aspiration in seven. Serious complications occurred in 11 patients, most commonly in those who had hypoxia or thrombocytopenia, but no morbidity occurred in 22 patients. The diagnostic yield allowed effective rational therapy to be selected for multiple potential pathogens in 12 patients and detected pathogens not suspected in six instances. The technique compares favorably to other invasive techniques for establishing definitive information in this group of seriously ill patients. PMID- 7471839 TI - Ejection fraction. What are its limits? PMID- 7471840 TI - Pathologic findings in a patient dying with ventricular tachycardia. PMID- 7471841 TI - Paroxysmal ventricular arrhythmias and familial sudden death associated with neural lesions in the heart. AB - A high school athlete with history of syncopal attacks died suddenly. Paroxysmal ventricular arrhythmias had been documented many times, as had at least one episode of ventricular fibrillation. A brother with similar history also had died suddenly and unexpectedly at about the same age. Except for the syncope and arrhythmias, they were both considered to be in good health. At postmortem examination, no significant extracardiac abnormalities were found, and the heart was normal on gross examination. The cardiac conduction system was the subject of special study. Focal inflammatory degeneration of small nerves and ganglia was found in various sites within the heart, including atrioventricular node, but were especially prominent in and around the sinus node. There was epicardial edema and thickening of the pericardium in that vicinity, but all of the pericardium elsewhere was normal. Some persistent fetal dispersion of the atrioventricular node was present. Ways are discussed in which these neural lesions may have contributed to the pathogenesis of paroxysmal arrhythmias and eventually sudden death. The possible etiology of the neural disease and the basis for its familial occurrence are considered. PMID- 7471842 TI - Chlorambucil in sarcoidosis. AB - Ten patients with biopsy-proven sarcoidosis with progressive disease were treated with chlorambucil alone or in combination with corticosteroids. Eight of these ten patients showed some degree of improvement which may be attributable to chlorambucil. The responsive patients showed beneficial effects within three months of starting chlorambucil therapy. It is concluded that chlorambucil may be worth a trial in patients with progressive disease unresponsive to corticosteroids or where corticosteroids are contraindicated. PMID- 7471843 TI - First pass radionuclide assessment of right and left ventricular ejection fraction in chronic pulmonary disease. Effect of oxygen upon exercise response. AB - Right and left ventricular ejection fraction (RVEF; LVEF) were determined in patients with severe chronic pulmonary disease (mean +/- SEM [FEV1 percent predicted 36 +/- 3%; PaO2: 64 +/- 3 mm Hg]), utilizing first pass radionuclide angiocardiography. RVEF and LVEF were measured at rest and again during upright bicycle exercise while patients breathed room air, and again during low flow oxygen (O2) administration. Mean RVEF was abnormal (less than 45%) at rest and did not increase with exercise while subjects breathed room air (44 +/- 2 percent vs 44 +/- 3 percent, P = ns), but improved significantly during exercise while patients breathed O2 (45 +/- 5 percent vs 51 +/- 3 percent, P less than .05). Breathing room air, RV exercise ejection fraction was abnormal (less than 5 percent increase in absolute RVEF) in 15 of 18 patients, but only 5 of 10 patients were abnormal during O2 administration. LVEF at rest was normal in all subjects. These data suggest: 1) RV exercise ejection fraction is abnormal in most patients with chronic pulmonary disease; 2) while low flow O2 does not alter RV performance at rest, it improves RV exercise ejection fraction in some patients. PMID- 7471844 TI - Response to inhaled metaproterenol and isoproterenol in asthmatic and normal subjects. AB - Pulmonary function tests were performed before and after inhalation of 1.3 mg of metaproterenol sulfate or 150 microgram of isoproterenol hydrochloride by 40 asthmatic and 40 normal subjects. PHysiologic measurements included spirometric testing, plethysmographic studies, and maximal expiratory flow-volume curves obtained after inhalation of air and a mixture of 80 percent helium and 20 percent oxygen. In the normal subjects, pulmonary function improved significantly after inhalation of both metaproterenol and isoproterenol. There was no significant difference in responsiveness to either bronchodilator drug. In the asthmatic subjects, pulmonary function also improved significantly after both bronchodilator agents. The sites of predominant bronchodilatation were similar in the asthmatic subjects after both metaproterenol and isoproterenol; however, bronchodilatation was better overall (P less than 0.005) and for most individual tests with metaproterenol. The greater efficacy of metaproterenol in asthmatic subjects but not in normal subjects can be explained by (1) different doses of the bronchodilator drug and (2) differing bronchodilator dose-response relationships in asthmatic and normal subjects. PMID- 7471845 TI - The diagnostic value of pleural fluid pH. AB - One hundred eighty-three patients had simultaneous blood and pleural fluid pH determinations. Thirty-six effusions were transudates, and 147 were exudates. In 46 effusions, the pleural fluid pH was less than 7.30; all 46 were exudates. A pleural fluid pH less than 7.30 was associated with the following six diagnoses: (1) empyema; (2) malignancy; (3) collagen vascular disease; (4) tuberculosis; (5) esophageal rupture; and (6) hemothorax. The results of pleural fluid pH determination are immediately available, narrow the differential diagnosis of the exudate, and may expedite patient management. The pH of pleural fluid should be measured whenever a diagnostic thoracocentesis is performed. PMID- 7471846 TI - Echo-phonocardiographic evaluation of obstruction of prosthetic mitral valve. AB - Echo-phonocardiograms of 64 patients with multiple types of prosthetic mitral valves (PMV) were evaluated. Six patients demonstrated findings suggesting valve obstruction: four had surgical confirmation of prosthetic obstruction, one reduced cardiac output and pulmonary hypertension but no prosthetic dysfunction at surgery, and one is asymptomatic. Three of four patients with confirmed obstruction had variable second sound to opening click intervals (A2-MO) with interrupted disc opening; two had interrupted disc closure with split closing clicks and three of four had reduced diastolic closure rate. In 58 clinically well patients with PMV, cycle-to-cycle A2-MO varied little: 0-10 msec in sinus rhythm and 10-25 msec in atrial fibrillation. Diastolic closure rates of five different types of PMV were similar: 21.6 mm/sec, (range 14-49). No patient had interrupted opening, closing or multiple closing clicks. Thus, delayed PMV opening or closure, altered A2-MO interval and double closing clicks are highly useful in detecting patients with obstruction of a variety of mitral prostheses. PMID- 7471847 TI - Guidelines for organization and function of hospital respiratory care services. Section on Respiratory Therapy, American College of Chest Physicians. PMID- 7471848 TI - Artificial intermittent ventricular parasystole with phase 4 protection. PMID- 7471849 TI - Mitral valve prolapse simulating left atrial myxoma. Noninvasive correlation with angiographic findings. PMID- 7471850 TI - Self-induced pulmonary granulomatosis. A consequence of intravenous injection of drugs intended for oral use. PMID- 7471851 TI - The influence of low frequency acupuncture on a demand pacemaker. AB - Various complications with demand pacemakers have been known to originate from their own sensing mechanisms. In these complications, electromagnetic interference is one of the well known causes of oversensing with demand pacemakers. The influence of low frequency acupuncture as a specific type of electromagnetic interference on a demand pacemaker was examined during a cervical operation. It was found that low frequency acupuncture is a cause of electromagnetic interference with demand pacemakers. Therefore, caution should be exercised when using the low frequency acupuncture on a pacemaker patient. PMID- 7471852 TI - Metastatic cardiac tumor demonstrated by 201thallium scan. AB - A patient with bronchogenic carcinoma had an electrocardiogram that was initially thought to be consistent with myocardial infarction and possible left ventricular aneurysm. A heart scan with radioactive 201thallium showed an area of sharply diminished uptake ("cold spot"); however, subsequent thoracotomy showed that this was caused by local pericardial and myocardial infiltration by tumor. This suggests a role for radioactive 201thallium in the detection of cardiac metastatic tumors. PMID- 7471854 TI - Incentive spirometry: the answer is blowing in the wind. PMID- 7471853 TI - Vasodilator therapy in primary pulmonary hypertension. PMID- 7471855 TI - Interpretation of tests of early lung dysfunction. PMID- 7471856 TI - Small airways disease: reproducibility of measurements and correlation with lung function. AB - Three measurements of small airway dimensions and a measurement of lesions in small airways were made in 32 human lungs obtained at autopsy in order to assess the intraobserver and interobserver variability of these measurements and their interrelationships. The same measurements were then made in 22 lobes resected in patients with known lung function. Small airway dimensions were smaller in surgically resected lobes compared to autopsy lungs suggesting persistence of muscle tone in the surgical specimens. PMID- 7471857 TI - Variability of the volume of isoflow. AB - Twenty healthy hospital workers produced maximal expiratory flow-volume curves before and after three vital capacity inhalations of an 80 percent helium and 20 percent oxygen mixture (HE+O2) in the morning and afternoon for four days during one week. Ten healthy trade union apprentices underwent the same tests, twice on one day and again one month later. Measurements made from curves (and their mean coefficients of variation) were: VisoV (105 percent) FVC (3 percent), FEF50% (6 percent), FEF75% (8 percent), the ratio of FEF50% breathing He+O2 to FEF50% breathing air (6 percent), and the ratio of FEF75% breathing He+O2 to FEF75% breathing air (9 percent). Differences among separate observers contributed significantly to the high variability of the VisoV. Diurnal changes and training effects over the week of study were not significant. In conclusion, VisoV is poorly reproducible compared with the FVC and expiratory flow rates at low lung volumes breathing air and He+O2. This should be considered when interpreting results in a given individual. PMID- 7471858 TI - Predominant site of airway resistance in chronic obstructive pulmonary disease. AB - The purpose of the present clinical research was to establish the major site of resistance and obstruction in patients with severe, symptomatic chronic obstructive pulmonary disease. We used a noninvasive technique by measuring airflow in a plethysmograph after patients breathed air and after breathing a mixture of 80 percent helium and 20 percent oxygen. In group 1 were 14 patients (mean age, 63 +/- 7 years [+/- 1 SD]) with bronchitis. The ratio of their forced expiratory volume in one second over the vital capacity (FEV1/VC) was 43 +/- 10 percent (mean +/- 1 SD), and their single-breath carbon monoxide diffusing capacity (Dsb) was normal (125 +/- 41 percent of the predicted value). In group 2 were 13 patients with emphysema (mean age, 60 +/- 10 years), whose FEV1/VC was 33 +/- 11 percent and Dsb was 55 +/- 14 percent of predicted. All patients had abnormally low increases in the maximum expiratory flow at midvital capacity after breathing the helium-oxygen mixture. The range in group 1 was -11 percent to +14 percent and in group 2 was -10 percent to +15 percent. Normal subjects in our laboratory show increases of at least +24 percent. This indicates that the limitation of airflow occurs primarily in the small (less than 2 mm in internal diameter) and not the large airways. PMID- 7471859 TI - Intravenous propranolol therapy for acute myocardial infarction in man: hemodynamic and serial creatine kinase assessment. AB - Propranolol was administered intravenously to 12 patients with presumed acute myocardial infarction in the attempt to limit infarct size. Patients' conditions were uncomplicated (heart rate greater than or equal to 60/min, systolic blood pressure greater than or equal to 100 mm Hg, mean pulmonary capillary wedge pressure mean [PCWP] less than or equal to 20 mm mercury). The aim was to produce beta-blockade that was early, complete, and continuous. Target loading dose was achieved in seven patients and full maintenance was achieved in six patients. The remaining patients received smaller loading or maintenance doses or both because of varying degrees of bradycardia, hypotension, or elevated mean PCWP. Myocardial CK release in the propranolol group was 2651 mIU/ml +/- 843 (mean +/- SE, n = 12) vs 2987 mIU/ml +/- 422 in 21 comparison patients, a difference not statistically significant. The time to CK plateau (completion of infarction) was related to total CK release in both propranolol and comparison patients. PMID- 7471861 TI - The role of isoproterenol in pulmonary artery hypertension of unknown etiology (primary): short- and long-term evaluation. AB - The experience derived from the administration of isoproterenol in six patients with pulmonary hypertension of unknown etiology (PAH-UE) is presented. The diagnosis was made after exclusion of other known diseases capable of producing hypertension in the pulmonary circuit. Catheterization was performed, and basal cardiopulmonary parameters, mean pulmonary artery pressure (PAP), pulmonary arteriolar resistance (PAR), cardiac index (CI), alveolar-arterial oxygen tension difference P(A-a)O2, and PaO2 were investigated. The effect of infusing 3 micrograms/min of isoproterenol into the pulmonary artery was studied in five cases. Isoproterenol was given sublingually to one patient who had previously received it intravenously; in another case it was given only sublingually. Significant P values (P less than .05) as a group were obtained, in relation to heart rate, CI, PAR, and mean PAP after isoproterenol. A favorable effect on the heart and lungs was seen in two cases, maintained for three years with sublingual isoproterenol with a favorable cardiorespiratory effect. Use of isoproterenol in PAH-UE is justified at present in those cases with a favorable cardiopulmonary response while no specific therapy is available. PMID- 7471860 TI - Familial type II hyperlipoproteinemia with coronary heart disease: effect of diet colestipol-nicotinic acid treatment. AB - Heterozygous familial type II hyperlipoproteinemia (F type II) is primarily manifested in hypercholesterolemia (due to low density lipoprotein-cholesterol [LDL-C] elevation) and premature coronary heart disease (CHD). We studied sequentially the effects of low cholesterol-low saturated fat-low simple carbohydrate diet; diet and colestipol, 30 g/day; and diet, colestipol, plus nicotinic acid (NA) 3 to 7 g/day on plasma cholesterol (Ch), LDL-C, triglyceride (TG), high density lipoprotein-cholesterol (HDL-C) and angiographically documented coronary arterial lesions of 32 F type II patients. Effective control of F type II resulted in arresting the progression of angiographically demonstrated coronary arterial lesions. PMID- 7471862 TI - Complications of bronchoscopy: comparison of rigid bronchoscopy under general anesthesia and flexible fiberoptic bronchoscopy under topical anesthesia. AB - A prospective study of 4,595 bronchoscopic procedures performed over four years (1975 to 1978) on 2,143 patients with various bronchopulmonary diseases is described. Of the 4,595 procedures, 1,146 were performed with a flexible fiberoptic bronchoscope (Olympus BF-5B2 or BF-B2) under topical anesthesia with tetracaine and procaine, and 3,449 procedures were performed with a rigid bronchoscope under general intravenous anesthesia with hexobarbital (Evipan) using a modified Sanders' technique to ventilate the patients. Complications occurred in 235 procedures (5.1 percent). Major complications that threatened the patient's life and required intensive medical treatment, surgical intervention, or resuscitative measures occurred in 51 procedures (1.1 percent); deaths occurred after 6 procedures (0.1 percent). A comparison of the complications of rigid bronchoscopy and flexible fiberoptic bronchoscopy revealed significantly higher rates of complications of fiberoptic bronchoscopy attributable to toxic effects of tetracaine and of complications of rigid bronchoscopy associated with insufficient general anesthesia. With rigid bronchoscopy, the number of major complications induced by diagnostic manipulations through the bronchoscope and the total number of major complications were significantly higher than with flexible fiberoptic bronchoscopy. PMID- 7471863 TI - Transcutaneous continuous wave Doppler ultrasound in the diagnosis of left atrial myxoma. AB - Six cases of left atrial myxoma were studied using transcutaneous, continuous wave, low-frequency Doppler ultrasound. The recordings, made from the suprasternal notch and the cardiac apex, showed characteristic deflections that coincided with tumor motion and provided a useful ancillary technique for noninvasively confirming the echocardiographic diagnosis of atrial myxoma. PMID- 7471864 TI - The bronchial leak squeak: a new sign for the physical diagnosis of bronchopleurocutaneous fistula. AB - We describe a unique physical diagnostic sign that we have observed in seven patients with bronchopleurocutaneous fistulas. Such patients have a highpitched squeak over the affected chest area during sustained Valsalva maneuver. We postulate that turbulence across the bronchial fistula due to high transbronchial pressure gradient during the Valsalva maneuver produces the squeaking sound. The pitch of the leak squeak sign is higher in smaller fistulas than in larger fistulas; decreases in intensity and increases in pitch occurred in two patients in whom the bronchial fistula slowly closed. The absence of the leak squeak sound in patients with spontaneous pneumothorax suggests that this sign can be used to differentiate central airways from alveolar air leaks. PMID- 7471865 TI - Bigeminy in ventricular interpolation. PMID- 7471866 TI - Running to death. PMID- 7471867 TI - Carcinoid tumor of the thymus associated with recurrent pericarditis. AB - A 49-year-old man with carcinoid tumor of the thymus presented with the clinical picture of acute pericarditis. Recurrent bouts persisted after removal of the tumor without other evidence to indicate tumor recurrence. There were no associated manifestations of an endocrine disorder. PMID- 7471868 TI - Cardiac rupture and ventricular septal defect in isolated right coronary artery disease. AB - A patient is described who had both ventricular septal defect and then cardiac rupture with death within 24 hours of the onset of acute posteroinferior myocardial infarction. At autopsy he was found to have single-vessel disease involving the right coronary artery. Isolated disease of the right coronary artery can produce unexpected, fatal mechanical complications. PMID- 7471869 TI - Pacemaker-induced hypotension. AB - We report a case of hypotension and decreased cardiac output resulting from artificial ventricular pacemaker rhythm. Hemodynamic deterioration was associated with loss of normal time relationships between atrial and ventricular contraction and paradoxical fall in systemic vascular resistance despite a low output state. The therapeutic implications of this unusual response are discussed. PMID- 7471870 TI - Transient superior vena cava syndrome due to propylthiouracil therapy in intrathoracic goiter. AB - Intrathoracic goiter is a rare cause of superior vena cava syndrome. We present the findings in a patient in whom the syndrome was precipitated by therapy with propylthiouracil and remitted on withdrawal of the medication. The superior vena cava syndrome did not recur on medical management, suggesting that surgery is not always indicated in this setting. PMID- 7471871 TI - Lethal occult pulmonary hemorrhage in drug-induced thrombocytopenia. AB - A patient with drug-induced thrombocytopenia who died because of massive pulmonary hemorrhage is described. The patient had a clinical picture of acute respiratory distress resembling pulmonary edema, but there was no hemoptysis. Chest x-ray films showed granular density in the pulmonary fields, and the electrocardiograms revealed a pattern of acute biatrial enlargement. The diagnosis was confirmed at autopsy. PMID- 7471873 TI - Tricuspid atresia with persistent truncus arteriosus. AB - Detailed pathologic anatomy of tricuspid atresia associated with type I persistent truncus arteriosus is described. These findings are correlated with clinical presentations. PMID- 7471874 TI - Pleural effusion: the thorn sign. Not a rare finding. AB - Eleven percent of an unselected series of 157 patients with pleural effusion showed a thorn-like protrusion of fluid at the lateral end of the minor fissure at some time during their illnesses. None of a series of 146 normal x-ray films of young patients showed this "thorn sign." The thorn sign is a useful radiographic finding of right-sided pleural effusion. PMID- 7471872 TI - Persistent postpneumonic pneumatoceles in children. AB - Ten children who had developed pneumatoceles during the course of pneumonia are described. Seven of the ten patients had persistence of pneumatoceles for 12 months or more. There was no evidence of an underlying disorder in any of the patients, and all recovered completely without complications. The etiologic agent was identified in two of the ten cases as Staphylococcus aureus. In our experience, a delay in the resolution of pneumatoceles for a period of one or more years after an episode of pneumonia is not an unusual occurrence in normal healthy children. PMID- 7471875 TI - Intrapleural positioning of esophagus for treatment of swallowing-induced arrhythmia. AB - Swallowing-induced atrial tachycardia is a rare phenomenon generally assumed to be caused by abnormal parasympathetic reflexes arising in the esophagus or pharynx. We describe a patient with intractable swallow tachycardia. Since certain features of the case suggested a mechanical rather than a reflex mechanism, he was treated by intrapleural repositioning of the esophagus to effect physical separation of esophagus and left atrium. Ten months after an uncomplicated procedure, the patient remains asymptomatic and free of arrhythmia. PMID- 7471876 TI - Left ventricular aneurysm, thrombus, and embolism. PMID- 7471877 TI - Oblique hilar tomograms in preoperative staging of carcinoma of the lung. PMID- 7471878 TI - Blood potassium measurements during CPR. PMID- 7471879 TI - Mitral regurgitation in ventricular premature contractions. PMID- 7471880 TI - Problem solving in occupational airways disorders. Cotton dust exposure as a case in point. PMID- 7471881 TI - Review of cotton dust control technology studies at North Carolina State University. Cultural, genetic, and ginning variations. PMID- 7471882 TI - A mortality study of workers in Lancashire cotton mills. PMID- 7471883 TI - The use of pulmonary function testing and questionnaires as epidemiologic tools in the study of occupational lung disease. AB - Pulmonary function testing and questionnaires are valuable tools in epidemiologic studies of occupational lung disease. Accurate equipment and standardized methodology are vital to obtain reproducible responses. For spirometry, the FVC and FEV show the least intrasubject variability and on questionnaires, occupational and smoking history are more reproducible than symptoms. The limitations of any method used to define a lower limit of normal should be kept in mind and, whenever possible, groups should be compared by use of the distribution of observations in the two groups--not just the prevalence of "abnormal" findings. PMID- 7471884 TI - Worker monitoring in byssinosis. PMID- 7471885 TI - The respiratory health of Quebec textile workers. PMID- 7471886 TI - Study of respiratory effect of short- and long-term cotton gin exposure. AB - No excess of obstructive airway disease was found in a group of 265 cotton gin workers when compared with other San Joaquin Valley agricultural workers. After an average of eight weeks' employment in San Joaquin Valley cotton gins, 125 workers showed no appreciable deterioration of pulmonary function compared with pre-employment measurements. In studies of pulmonary function during a workshift, cotton gin workers showed slightly greater mean decrements than control agricultural workers. These differences did not reach a level of statistical significance and were lower than those usually found in byssinosis. The unusual temporal pattern of employment in cotton gins in California precludes a simple approach to diagnosis by symptoms. No correlation was found in this study between symptoms of byssinosis and objective decrements in FEV1. The questionnaire as proposed by the Cotton Dust Standard was found to be of no value in detecting reactors in this study of gin workers. PMID- 7471887 TI - The relation of lung function to subsequent employment status and mortality in cotton textile workers. PMID- 7471888 TI - Worldwide problems of byssinosis. PMID- 7471890 TI - Comparative prevalence and severity of emphysema and bronchitis at autopsy in cotton mill workers vs controls. PMID- 7471889 TI - A purified extract from cotton bracts induces airway constriction in humans. PMID- 7471891 TI - Cotton dust and complement in vivo. AB - Cotton grown in Stoneville, Mississippi and harvested by three different methods was examined for anticomplementary activity in human serum in vivo. Subjects exposed to cotton dust in a model cardroom for six hours showed changes in C3c and CH50 after exposure to closed boll harvested, bract removed cotton dust when compared with a group of normal, unexposed controls. Decreases in C3c, :C4, and C3A were associated with decreases in FEV1 after exposure to closed boll harvested bract intact cotton dust. The data from this preliminary study suggest in vivo associations between inhalation of cotton dusts and complement which appear to be independent of endotoxin contamination. These findings suggest a relationship between cotton dust inhalation and complement-mediated respiratory impairment. PMID- 7471892 TI - Mortality of employees of two cotton mills in North Carolina. PMID- 7471893 TI - Studies on the etiology of byssinosis. PMID- 7471894 TI - The pathology of byssinosis. AB - In 1768, Morgagni gave the history and post=mortem findings in a man of 40 who had worked with hemp and found no abnormality in the lungs. It was the absence of gross pathology in the lungs of those who died severely disabled by byssinosis that led to our research into the disease. We see about 107 deaths a year of those with byssinosis who have been examined in life and have been given varying assessments of disablement. This article compares the pathologic findings in the lungs of a number of subjects with byssinosis with their ventilatory capacity in life and their smoking, occupational and clinical history. Emphysema is not an invariable finding. Interstitial fibrosis, which might be expected if the etiology of byssinosis was akin to farmer's lung, is not found. There is no characteristic lung lesion as is seen in the pneumoconioses. Cotton fibers are not found in the lungs and "byssinosis bodies" are not present in a majority of the cases. The pattern that emerges for byssinosis is that of asthmatic bronchitis. PMID- 7471895 TI - Pulmonary function response to dust from standard and closed boll harvested cotton. PMID- 7471896 TI - Pulmonary function and symptoms in herbal tea workers. PMID- 7471897 TI - The relationship of acute respiratory response and chronic respiratory symptoms in byssinosis. PMID- 7471898 TI - International conference on byssinosis. Overview of environmental evaluation and control. PMID- 7471899 TI - Modern plant dust control techniques. PMID- 7471900 TI - Vegetable ingredients of dusts in textile and nontextile cotton industries. AB - The objective was to determine if garnetting and raw cotton dusts differed in botanical composition. Estimates of dust composition were based on the contents of gross trash in cotton raw materials and the potential of each gross trash component to be converted into less than 10 mum particulate by a laboratory abrasion test. All types of garnetting dusts are predicted to contain a lower percent content of leaflike material than raw cotton dust. Garnetting dusts arising from linter-polyester blends are estimated to contain only 20 to 25 percent of the concentration of leaflike ingredients predicted to be present in raw cotton dusts. The low prevalence of byssinosis reported in the garnetting industry may be explained, in part, by the relatively low percent content of leaflike particulate in this cotton dust. PMID- 7471901 TI - Presence of antibacterial agents in blood units collected from healthy voluntary donors. AB - 10,991 sera collected from voluntary blood donors were tested during a 1-year period (1979) for the presence of antibacterial drugs. Antibacterial activity was found in 0.25% of them, and could be related to beta-lactam antibiotics and to cotrimoxazole. The implications of these findings are discussed. PMID- 7471902 TI - Effect of bouvardin (NSC 259968) on the growth characteristics an nucleic acids and protein syntheses profiles of P388 leukemia cells. AB - The plant product Bouvardin (BVD; NSC 259968) exhibited high activity against P388 murine leukemia and B16 melanoma, but it was not effect against various other tumor models. BVD inhibited the synthesis of all the macromolecules, namely protein, DNA and RNA, in P388 cells in vitro. Protein synthesis was most susceptible to the drug action. However, DNA and RNA synthesis were affected to a lesser extent. Further studies revealed that BVD did not inhibit protein degradation in P388 leukemia cells. PMID- 7471903 TI - The effect of chloramphenicol on erythropoiesis in X-irradiated mice. AB - Mice irradiated with a dose of 4,78 Gy X-radiation were treated 3 or 5 days with a thrice-daily dose of chloramphenicol of 160 or 320 mg/kg. Observation of the incorporation of 59Fe into the heme fraction suggested that chloramphenicol has a depressive effect on erythropoiesis in the spleen of nonirradiated animals, and clearly proved this in irradiated ones. Depression in the bone marrow of nonirradiated animals was not observed, in irradiated animals a depressive effect was shown, though less strong than in the spleen. As benefits these findings there is an accentuation of postirradiational anemia after chloramphenicol treatment of irradiated animals, and signs of incipient anemia in nonirradiated animals. All the effects mentioned were repaired within 14 days of the end of chloramphenicol treatment. PMID- 7471904 TI - Pharmacokinetics of Metronidazole in pregnant women. AB - Pharmacokinetic characteristics of metronidazole were studied in 19 pregnant women with Trichomoniasis urogenitalis who had been hospitalized for legal abortion. All patients received 250 mg or 1.0 g metronidazole (Vagimid) per os in a single or multiple dose. Peak levels were reached 1-2 h after application. Mean serum levels of metronidazole were somewhat lower in gravidae than those in nonpregnant women. The mean elimination half-life values were 5.7 and 7.7 h after intake of 250 mg and 1.0 g metronidazole, respectively. As suggested by our pharmacokinetic results, the dosage regimen for treatment of anaerobic bacteria infection or parasitic infection in pregnant women can be the same as in nonpregnant patients. PMID- 7471905 TI - Binding (in vitro) of some antimitotic isatin derivatives to human serum albumin. AB - The binding of isatin and its mustard N-Mannich base, considerably biologically active compounds, to human serum albumin has been studied by equilibrium dialysis and ultrafiltration. The influences of ligand and macromolecule concentration, temperature and pH of the incubation medium have been demonstrated. The Scatchard plot of isatin binding to albumin shows a biphasic curve which indicates the presence of at least two different binding sites on albumin molecule. One site with a higher affinity, K1 = 2.25 X 10(3) M and n1= 25, and the other site with a lower affinity, i.e. higher capacity. In the cases of mustard Mannich base we could demonstrate the same type of curve, K1 = 2.20 X 10(5) M and n1 = 1.0, whereas another site has a lower affinity and greater number of binding sites. PMID- 7471906 TI - Bioassay of clindamycin in bile. AB - This study compares different preparations of standard solutions for determining clindamycin in bile. Pooled human serum, ox bile, 10% human bile, and 0.1 M phosphate buffer were compared, and pooled human bile was serving as reference standard curve. The use of pooled human serum and ox bile resulted in a considerable error of overestimation of clindamycin in bile. An error of underestimation occurred when 10% human bile or 0.1 M phosphate buffer was used as a standard solution. It is concluded that none of the different preparations tested are acceptable substitutes for human bile. PMID- 7471907 TI - Comparison of direct and standardized antibiotic susceptibility testing in bacteriuria. AB - In clinical isolates of bacteria from urine, the antibiotic susceptibility was tested with a direct disc diffusion method in which the inoculum was not primarily standardized. The results were compared with those from parallel tests using standardized inoculum. Of a total 2,272 urine specimens, 700 were accepted for direct susceptibility testing, 189 were not accepted because of mixed culture or indistinct inhibition zones. The overall concordance between the direct and the standardized tests was 95.8% (2,800 observations with 118 discordant results). The direct diffusion method used in this way is time saving and economical and can reliably be used in a clinical microbiological laboratory serving mainly outpatients. PMID- 7471908 TI - Primary prevention of child mistreatment: meeting a national need. AB - Greater emphasis on preventive efforts in the growing problem of child abuse and neglect has become a national necessity. This article presents a framework for action on federal, state and local levels. PMID- 7471909 TI - To silence one's self: a brief analysis of the literature on adolescent suicide. AB - This paper discusses and analyzes the etiology and treatment of suicidal adolescents from an ecological point of view, involving three major perspectives biophysical, psychological and sociological. PMID- 7471910 TI - Foster parent input: a crucial element in training. AB - A demonstration program in the training of foster parents was based on foster parents' responses to a questionnaire dealing with preferences as a topics and trainer personnel. PMID- 7471911 TI - The Preschool Behavior Rating Scale. PMID- 7471912 TI - Cerebral injury in child abuse: a changing profile. AB - This report of 21 patients with cerebral injury resultant from child abuse, identifies a broad profile of central nervous sequelae from such injuries, 6 patients were admitted with whiplash shaken infants syndrome, all with severe signs of CS dysfunction. 3 children presented with acute cerebral confusions, 2 with severe cord injuries with resultant paresis and 10 with skull fractures or miscellaneous injuries. Computer tomography has proved to be of great assistance in precise identification of site and size of subdural/intracerebral hematomas in abused children and in identifying posterior fossa lesions and providing longitudinal assessment of sequelae of cerebral trauma. Also the role of the Child Protection Team in management of such acutely injured children is defined. PMID- 7471913 TI - Chemotherapy of pediatric posterior fossa tumors. AB - Results of chemotherapy of recurrent posterior fossa tumors in children are presented. Cerebellar astrocytoma, ependymoma, and brain stem glioma have all shown objectively determined responses to treatment with nitrosourea compounds (BCNU or CCNU). Medulloblastoma has shown objectively determined response to various chemotherapeutic agents; but in our experience the best responses have been obtained with a combination of a nitrosourea (CCNU), vincristine, and procarbazine. Although chemotherapy after recurrence has significantly prolonged the life of the patients, cure has not been achieved. Theoretical considerations regarding the failure of initial therapy and the inadequacy of subsequent chemotherapy are discussed. PMID- 7471914 TI - Evaluation of complicated migraine in childhood. AB - We report on 3 patients with complicated migraine of childhood. Family history was positive for migraine in each case, and all experienced the acute onset pf neurologic deficits in associated with throbbing hemicranium. Abnormalities of cerebral blood flow determinations, electroencephalograms and cerebral arteriograms in addition to persistent neurologic sequelae were demonstrated. We propose that complicated migraine of childhood is not an entirely benign symptom complex. PMID- 7471915 TI - Retroperitoneal neuroblastoma metastatic to brain. Report of a case and review of the literature. AB - Brain metastases from extracranial neuroblastoma are rare. They are generally produced by direct extension from metastatic lesions of skull or dura. Very few cases with primary cerebral metastases have been described. We reviewed all autopsy cases of extracranial neuroblastoma at Children's Memorial Hospital in the 12-year period from 1976 to 1978. Of 27 cases in which the brain was examined, 14 had intracranial involvement but only one presented parenchymal metastases in the absence of skull or dural involvement. This case is reported in detail and the literature is reviewed. PMID- 7471916 TI - Development of visual organization: the perception of subjective contours. AB - This study examines the development of infants' sensitivity to the organization of a subjective-contour stimulus array. 5- and 7-month-olds were sequentially shown 3 stimulus arrays of elements, only 1 of which was capable of producing subjective contours. Only the orientational relations among elements was varied. An infant habituation control procedure was used to test infants' abilities to discriminate these arrays. The results indicated that (1) only 7-month-olds showed consistent differential responsiveness to changes from an illusory array to a nonillusory array or vice versa, (2) 5-month-olds showed a weaker tendency to respond similarly and only when they had prolonged experience with the illusory array, and (3) neither age group showed much response recovery to a change from one nonillusory array to another. These findings are interpreted as indicating that infants can perceive subjective contours. However, the age of this accomplishment probably varies with both the characteristics of the array and the abilities of the observer. PMID- 7471917 TI - Developmental differences in visual backward masking. AB - 2 experiments were conducted to examine developmental differences in visual recognition masking when verbal encoding and response demands are minimized. Precautions were taken in the experiments to ensure the implication of the central visual system in the backward-masking component of each experiment. Sampling was conducted across the kindergarten, third-grade, sixth-grade, and college levels. In the first experiment, no-mask target recognition was set at 100% correct. The results indicated systematic grade differences in backward masking such that older children and adults showed more rapid recognition improvement across stimulus onset asynchronies (SOAs) than did young subjects (i.e., a grade x SOA interaction). Although this finding could be interpreted to suggest developmental differences in the rate of visual information processing, such an interpretation would be inappropriate. The problem is that the masking functions for all grade levels reach asymptote at 100% correct (i.e., a ceiling effect). Thus, the grade X SOA interaction could have been produced for reasons other than rate-of-processing differences (e.g., developmental differences in the quality of stimulus reception). Thus, experiment 2 was designed to assess this possibility by evaluating developmental differences in visual backward masking when no-mask target recognition was set at the 75% level for the grades sampled. The grade x SOA interaction observed in experiment 1 was not observed in experiment 2. The results from the studies indicate that, when the ceiling effects in no-mask target recognition are removed, the visual information processing rates for children and adults can be considered equivalent. PMID- 7471918 TI - Infant saccadic eye movements to visible and previously visible targets. AB - Recent evidence has shown that 1- and 2-month-old infants localize a peripheral target by means of a series of saccades. Step size in a series was directly related to target distance, but within a localizing series did not vary appreciably as the line of sight approached the target. The current study examined whether, during localization of a peripheral target, step size and number of steps were determined prior to the first saccade in a localizing series. 2-month-olds viewed peripheral targets which either remained on (continuous condition) or were extinguished (interrupted condition) prior to target localization. It was found that on the majority of trials a series of saccades was made toward the target hemifield. In both conditions the size of each saccade in a multiple saccadic series was approximately equal within a given trial, but varied with initial target distance. Multiple saccades on both continuous and interrupted trials were similar in form, differing only in the total extent of eye rotation. This suggests that a similar oculomotor program was operating under both conditions. PMID- 7471919 TI - Early word meanings: perceptually or functionally based? AB - The issue of whether early word meaning is based on perceptual (Eve Clark) or functional (Katherine Nelson) features has not been satisfactorily resolved by previous research. The present experiments addressed this issue by presenting young children (2- and 3-year-olds) with novel objects in which perceptual and functional features varied independently. Given choices of how to sort novel artificial objects varying in both aspects, children in experiments 1 and 2 chose perceptual features with few exceptions. Experiments 3 and 4 presented the same objects to children in a concept-learning task, where nonsense labels were to be learned for perceptually or functionally based categories; the latter were much more difficult. Experiment 5 was an extension of the first 2 experiments, except that more familiar objects were employed; comparable results were found. All of the results support 1 conclusion: early conceptualizations and word meanings are perceptually based when perceptual and functional features are independently available. PMID- 7471920 TI - The short-term and long-term amelioration of academic and motivational deficiencies among low-achieving inner-city adolescents. AB - 56 low-achieving seventh-grade students in a predominantly black, inner-city junior high school were randomly assigned to one of four groups (SMART, PUSH, Standard Reading, or Nonintervention). The highly structured, school-based SMART program employed nonprofessional tutors to improve the academic skills and attitudes of the adolescents. The home-based PUSH program involved parents in restructuring the home environment to foster educational development. The Standard Reading group was the usual school reading class, and the Nonintervention group did not receive any special remediation. A battery of standardized educational and attitudinal tests was administered to each student (a) immediately before assignment to groups, (b) immediately after termination of the 10-week programs, and (c) at 6-month follow-up. Results from the 4 x 3 analysis of variance clearly demonstrated the short- and long-term efficacy of the SMART program and the long-term efficacy of the PUSH program. Each produced substantial increases in both the acquisition of fundamental academic skills and achievement motivation. Implications of these findings and directions for future research are discussed. PMID- 7471921 TI - Maternal stimulation and infant exploratory competence: cross-sectional, correlational, and experimental analyses. AB - Research on maternal influence on infant development is criticized on the basis of its limited ability to make strong causal claims. 2 observational studies are then presented; 1 details, cross-sectionally, developmental changes in strategies of maternal stimulation and infant exploration and the interrelationship of these domains of parent and child behavior; the other tests, experimentally, the hypothesis that maternal stimulation fosters infant exploratory competence. Results of the first study reveal: (1) a linear increase in mother's verbal attention-focusing behavior and a curvilinear trend in physical attention focusing strategies between 9 and 18 months; (2) linear increases in a variety of measures of infant exploration; and (3) positive associations between naturalistically observed maternal stimulation and infant exploratory competence. The experimental study demonstrates that infant exploratory skill, as measured both naturalistically and during a semistructured, free-play assessment, is positively influenced by the enhancement of maternal stimulation. These latter results are discussed in terms of infant stimulation programs. PMID- 7471922 TI - Development of cardiac and behavioral responses to a three-dimensional toy stimulation in one- to six-month-old infants. AB - Concomitance between development of perceptual-motor behavior and cardiac-somatic association was evaluated in 1-6-month-old infants. The behavior was elicited by presenting 3-dimensional objects to 50 infants divided into 5 age groups. Each infant was presented with a sequence of 5 commercial toys on 2 immediately succeeding occasions. Each toy stimulation lasted 30 sec and was divided in 2 phases: distal and proximal to the infant. Cardiac responses during the distal phase (first 15 sec) differed as a function of maturing capability of infants to touch and grasp the toys when they subsequently became proximal (last 15 sec). In younger infants (aged 1-3 months), heart-rate (HR) response was consistently decelerative and indexed orienting and attention to the object. In older infants, HR response shifted to acceleration while they fixated the distal toys, before they could actually reach them. This difference in cardiac reaction was attributed to differing stages of perceptual-motor maturation which permit different transactions with external events: somatic activity (acceleration) or information-getting activity (deceleration). PMID- 7471923 TI - Infant wariness toward strangers reconsidered: infants' and mothers' reactions to unfamiliar persons. AB - In this study, the interpretation of infants' responses to approaching strangers as "wariness" or "fear" was questioned by comparing infants' behaviors toward strangers with adults' behaviours toward strangers. 24 8-month-old infants and their mothers were observed in a standard laboratory situation. Both infants and mothers were approached by female strangers with a fast, 12-sec approach and a slow, 40-sec approach. Mothers' and infants' behaviors were coded at 4 distances during each approach. Behaviors coded were smile, alert face, quizzical look, frown, looks at mother/infant, looks at stranger, and averts gaze. Mothers showed significantly more behaviors typically labeled "wary" than did infants, particularly as the proximity of the stranger increased. These findings suggest that wary responses toward strangers are not unique to infants and may be more characteristic of the situation than of the developmental level of the individual. PMID- 7471924 TI - The relationship between out-of-home care and the quality of infant-mother attachment in an economically disadvantaged population. AB - The effects of routine daily separations occasioned by out-of-home care on the formation and maintenance of infant-mother attachment relationships were examined in a population of economically disadvantaged mothers. 3 groups were constituted on the basis of the time in the infant's life when out-of-home care began: (1) before 12 months; (2) between 12 and 18 months; (3) home-care controls. The infant-mother pairs were observed in the Ainsworth strange situation at both 12 and 18 months, and were classified as secure, anxious-avoidant, or anxious resistant. Because previous research has implicated the psychological accessibility of the mother to the infant in the development of anxious-avoidant attachments during the first year of life, the hypothesis that physical inaccessibility due to out-of-home care would also be associated with anxious avoidant attachments was tested. The data support this hypothesis. At 12 months 47% of the infants whose mothers had returned to work/school were classified in the anxious-avoidant group, while the other 2 groups did not differ significantly in the proportions of infants assigned to the 3 attachment classifications. At 18 months, differences among the 3 work status groups also showed a large portion of anxious-avoidant infants (41%) in this early working group. However, infants whose out-of-home care began after 12 months did not show an increase in the proportion of anxious attachments. Additional analyses of variables related to mother's return to work indicated that single mothers were more likely to return to work/school, that mothers who worked reported higher levels of life stress than mothers who stayed home with the infants, and that, by 18 months, both anxious-avoidant and anxious-resistant attachments were also associated with non intact families. PMID- 7471925 TI - Training in social competence: the effect on mothers, fathers, and infants. AB - The effects of a parent-training program designed to increase parental competence to assess, predict, elicit, and contingently, respond to infant behavior and to increase the infant's contingent responses to the parents' behavior were assessed. 19 parent couples and their 4-12-month-old infants were randomly assigned to the training group or the control group. Raters, unaware of the status of the parents, observed and rated the parents' behavior during a 2-hour home observation. Separate raters, unaware of the hypothesis, then rated a 20-min videotape of the parent-infant interactions in the home. Parents independently completed a questionnaire. Training was found to increase both the parents' and the infants' competence in the parent-infant dyad. Most important, a reciprocal relationship between increases in the trained fathers' interactions and decreases in the trained mothers' interactions was found, indicating that the triad may be the crucial unit for studies of parental competence. PMID- 7471926 TI - Structural aspects of maternal speech to infants reared in poverty. AB - Mother-infant dyads (N = 27) were seen as part of a longitudinal study of lower socioeconomic-status infants at high but varying risk for mild mental retardation. A 20-min unstructured interaction session was videotaped in a laboratory setting when the infants were 6 months old. Transcriptions were made of mothers' speech to their infants. Measures of maternal language included sentence form, amount of speech, and syntactic complexity. It was found that the proportion of imperatives, but not the amount of maternal speech, was positively correlated with a composite measure of risk status (High Risk Index) and negatively correlated with maternal education and WAIS IQ. Similar to previous findings for maternal speech to older infants, syntactic complexity was not significantly related to social risk indices. In terms of descriptive aspects of the data, there was a high degree of variability on the language measures indicative of wide individual differences in maternal language style. PMID- 7471927 TI - The processing of tachistoscopically presented visual stimuli by five-month-old infants. AB - 5-month-old infants were presented visual patterns for 100 msec followed by a 100 msec patterned masker at intervals of 0, 250, 500, and 2,000 msec after the offset of the stimulus. This procedure was repeated 45 times. Only infants in the 2,000-msec stimulus-masker interval condition significantly fixated a novel stimulus longer than the familiar stimulus. These results suggest that visual processing in infants is quite slow relative to that in older humans. PMID- 7471928 TI - Intersensory function in newborns: effect of sound on visual preferences. AB - The results of this study indicate that newborn infants' fixation of a graduated series of visual stimuli comprised of a low (1), intermediate (4 or 16), and a high (64) number of cubes is significantly different in the absence and presence of sound (white-noise bursts). Relative to the no-sound condition, sound resulted in the infants' tendency to look more at the low-intensity visual stimulus and less at the high-intensity visual stimulus. These results provide support for the idea that the newborns' optimal or preferred amount of stimulation is based on the total amount or intensity of stimulus input, regardless of whether this is contributed to by stimulation from one or more than one modality. PMID- 7471929 TI - Preschool children's ability to coordinate spatial perspectives through language and pictures. AB - To determine preschool children's ability to utilize language in spatial problem solving, 64 children (equal numbers of boys and girls, 3- and 4-year-olds) were asked to identify another's view either verbally or by picture selection. The results indicate that verbalization leads to substantially more correct responses and girls were significantly better than boys across both response modes. The results imply that, despite the claims of several theorists, linguistic processes are accessible for problem solving at an early age. PMID- 7471930 TI - On the biological basis of sex differences in aggression. AB - The prevalence of aggression as a predominantly male behavior pattern has often been noted. The nature and perceived extent of sex differences in aggression have recently revived interest in speculation that such differences are attributable to factors inherent in the biological basis of sex differentiation. This paper will critically examine the empirical and theoretical basis for Maccoby and Jacklin's contention that males are biologically predisposed toward aggressive behavior. The literature to be examined includes: (1) cross-cultural studies of children's behavior, (2) the behavior of nonhuman primates, (3) sex hormones and aggression, and (4) early learning influences in child development. Contrary to Maccoby and Jacklin's assertions, these first 3 research traditions are found to indicate no biological predisposition toward aggression in human males. Furthermore, the pattern of existing evidence suggests that the gender-dimorphic nature of aggression is reliably observable in children's spontaneous behavior only after the age of 5 years. This finding presents great difficulty for the biological theory as proposed by Maccoby and Jacklin. Suggestions are presented for an interactional theory of the etiology of observed sex differences in adult aggression. PMID- 7471931 TI - Sex differences in aggression: a rejoinder and reprise. AB - A meta analysis of observational studies of peer-directed aggression by children aged 6 and younger yields a highly significant sex difference. Out of 32 studies, z values reflected higher male aggression in 24, no difference in 8, higher female aggression in none. Furthermore, boys' aggression is most often displayed in the presence of male partners. Evidence is presented that the sex difference is probably not merely an artifact of higher rates of male activity or social interaction. Existing cross-cultural evidence also shows higher rates of male aggression, as does most of the work on free-living primates. Specifically, the 3 observational studies of chimpanzees show considerably more aggression in males. Evidence for a hormonal contribution to male aggression is clear in animals and inconclusive in human beings, although the existing human findings are consistent with such a contribution. Recent evidence on the differential socialization of boys and girls supports our earlier view: that boys do not receive more reinforcement for aggression than girls, and that rates of punishment are also similar once the differential base rates in aggression are taken into account. The role of self-socialization (including choice of same-sex models) is discussed, and the view is expressed that this probably depends upon the development of certain cognitions about sex identity which normally do not develop until a later age than the age at which a consistent sex difference in aggression first appears. PMID- 7471932 TI - The development of perception and recognition of objects. AB - In this paper it is argued that the development of object perception in infancy involves the detection of structural invariants. Such detection is best understood in the context of dynamic events. Therefore, the role of continuous optical transformations in the infant's learning is emphasized. The effects of object motion and the infant's own activity are discussed in detail. PMID- 7471933 TI - The methodology of home-based intervention studies: problems and promising strategies. AB - Research on home-based interventions directed toward promoting positive interaction between mothers and their young children has become a less active field in spite of evidence that some programs sustain meaningful improvements in the lives of mothers and children. The purpose of this paper is to clarify some of the methodological problems of home-based intervention research and to suggest future directions. Several major projects are discussed to illustrate different perspectives, trade-offs, and promising strategies for dealing with key issues: (a) congruence between goals and methods, (b) sample selection and retention, and (c) measurement and design. The authors suggest that designing home-based interventions as longitudinal development research (rather than outcome evaluations) can provide valuable knowledge about ecological factors that affect the development of competence in parents and children and about effective ways to support families. PMID- 7471934 TI - [The distribution of authority in the hospital and the responsibility of the leading division chief]. PMID- 7471935 TI - [Experimental study of the optimization of the hip endoprosthesis]. AB - Clinical and laboratory investigations have resulted in a new design for the shaft prosthesis of the "St. Georg" model total hip prosthesis, the purpose of which was to minmize the risk of biomechanical complications. The original method of attachment and the same material for the limb were retained for our investigations. It was possible to test the bending strength, the length, and the geometry of the shaft as well as how structure of the bearing surface of the collar influences the force flow within the compound system. The distrubances produced in physiological force flow produced on insertion were also investigated. The results obtained enabled us to design a new shaft prosthesis. PMID- 7471937 TI - [Treatment of soft tissue infections with gentamycin-polymethylmethacrylate bead chains]. AB - Local antibiotic treatment with Septopal chains can also provide early control of soft tissue infections, provided the operative procedure is adequate. Of 80 patients treated this procedure was successful in 73% after the first operation and in 24% after reoperation. During the planning of treatment attention should be paid to the special requirements of this specific therapy; certain septic surgical methods have to be modified. Therefore limited extension of gentamycin diffusion has to be considered, and primary wound healing - if necessary by means of Epigard - should be attempted. PMID- 7471936 TI - [Surgical treatment of instable per- and subtrochanteric femure fractures with the 130 degrees double-T-plate. Clinico-experimental report]. AB - 1. Operative treatment of instable inter- and subtrochanteric fractures with a new 130 degrees double-T plate allowed, in spite of high-risk patients (age mean = 85 years, multimorbidity approximately 92%), full maximum stress by the third day after the operation and a shorter time of hospitalisation of 22.5 days. Lethality was 12.2%. - 2. Load experiments showed that the double-T-profiled 130 degrees plate with increased lateral moment of resistance could support a weight five times greater than that supported by 130 degrees implants with transversal blades, and had a resistance to bending two-and-a-half times greater than hip compression screws. In an 'intertrochanteric' position the plate could carry 3000 N and in a 'subtrochanteric' position 1000 N without permanent deformity. - 3. A review was carried out in 62.6% of the patients 6-32 months after double-T-plate osteosynthesis. Acceptable results were revealed in 80.7%. Of the 98 patients reviewed 44% had complete freedom of movement, 34.6% walked with a stick, 15.4% required crutches and 6% were unable to walk. PMID- 7471938 TI - [Amputation of the lower leg with formation of a musculo-cutaneous flap in vascular diseases]. AB - A report is presented on leg amputations with formation of a posterior musculocutaneous flap, as used since 1970 at the authors' clinic for vascular diseases. A total of 87 patients were operated on. In 74.7%, the amputation stump healed per primam; in 88.7%, movements of the knee joint were completely preserved. Rehabilitative walking on the prosthesis is possible very soon after the operation. The described method of amputation may also be recommended for patients with extensive loss of circulation of the lower extremities, including cases complicated by diabetes. PMID- 7471939 TI - [Surgical treatment of extensive cavernous hemangiomas of the face after arterial embolization]. AB - Six patients with voluminous cavernous hemangiomas on the face are reported. After devascularization according to the method of Djindjian the hemangiomas were removed surgically. The time interval between embolization and operation ranged from 2 to 7 days according to the hemodynamic situation of the angioma. The postoperative results are reported. PMID- 7471940 TI - [Percutaneous removal of intravascular fragments from infusion, angiography and cerebrospinal fluid drainage catheters with the loop-snare technic]. AB - Intravascular loss of catheter fragments (for infusion, angiographic, or CSF shunting purposes) represents a serious complication. There is a high morbidity rate and even mortality when the foreign body remains within the vascular system. Nonsurgical percutaneous removal of 16 catheter fragments from the venous side and 1 Gianturco coil from the abdominal aorta by use of a catheter sheath introducer system and the loop-snare technique is reported. If there is no local thrombosis, fragment recovery is relatively easy. Surgical intervention should be resorted to only when percutaneous removal turns out to be impossible. PMID- 7471941 TI - [Surgery of esophageal cancer]. AB - In the last 12 months 40 patients underwent surgery for carcinoma of the esophagus. In 18 cases the tumor was located in the lower third of the esophagus. An abdomino-thoracic approach was employed and a 2/3 resection of the esophagus and an esophagogastrostomy were performed. The resected area was bridged by an isoperistaltic gastric tissue tube and anastomosis was carried out using the EEA stapler. A telescope antirefluxplasty was performed to protect the anastomosis and to prevent a gastroesophageal reflux. One patient died of pneumonia. In all cases a secure anastomosis was achieved and in most cases gastroesophageal reflux was prevented. PMID- 7471943 TI - [Intraoperative marking of the radial nerve]. PMID- 7471942 TI - [Discopathy of the sternoclavicular joint]. AB - Among the joints of the shoulder region, the sternoclavicular joint (SJ) is under the most stress. Dragging pains emanating towards the neck and the lateral parts of the shoulder are typical complaints, although swelling of the SJ is rarely observed. Degeneration of the fibrocartilagenous disc should be considered when swelling is present, especially in the case of recurrent pains lasting for several months and swelling excluding inflammatory diseases, rheumatism and degenerating arthrosis. Above all we are interested in examining discopathy of the SJ joint and, in case of unsuccessful conservative measures, effective surgical therapy. We recommend the opening of the SJ, the resection of the degenerated disc and, finally, a tight capsulorrhaphy. Six patients operated in this way showed very positive post-operative results. PMID- 7471944 TI - [Blunt abdominal trauma and perforation by an intestinal foreign body]. PMID- 7471945 TI - [Implant and foreign body detector for surgery]. PMID- 7471946 TI - [Kuntscher nail extraction apparatus]. PMID- 7471947 TI - [Improved suspension for the Steinmann nail extension on the Weber table]. PMID- 7471948 TI - [An instrument for the blunt dissection of the esophagus]. PMID- 7471949 TI - [Surgical implants - risks and safety]. PMID- 7471950 TI - [Necessity and importance of explanation before surgery]. PMID- 7471951 TI - [Hemorrhagic diathesis and surgical intervention: emergency diagnosis and preoperative therapy]. PMID- 7471952 TI - [Intra- and postoperative measures in hemorrhagic diatheses]. PMID- 7471953 TI - [Effectiveness of continuous drainage in diffuse bacterial peritonitis]. AB - From 1975 to 1977 we carried out postoperative, continuous peritoneal lavage of th abdomen in 41 patients with diffuse bacterial peritonitis. Although the peritonitis was arrested more frequently and earlier, comparison to 71 patients treated conventionally since 1970 showed an increase of wound complications and a prolongation of hospital stay from 25 to 44 days. The higher frequency of complications led to increased lethality from 42% to 54%. The harm by continuous peritoneal lavage out-weights the advantages, except for stercoraceous peritonitis. PMID- 7471954 TI - [Congenital esophageal cysts]. AB - Congenital cysts of the esophagus are rare tumors of the mediastinum. They are classified as foregut-cysts. Most of them are found accidentally on routine chest X-ray. They are seldom diagnosed preoperatively. Exstirpation of the cysts presents no major problems. Five patients with congenital esophageal cysts are presented. PMID- 7471955 TI - [Annular pancreas - complications and therapeutic problems]. AB - Five cases of annular pancreas with symptoms and signs of high duodenal obstruction are reported. The diagnosis was established during the operation, and four of five patients were complicated by duodenal ulcer and a marked relapse of cholangitis. The results and the operative procedures are discussed. PMID- 7471956 TI - [Hemorrhoidectomy. Comparison of the Milligan-Morgan and Parks surgical methods]. AB - In 230 cases of patients operated on for hemorrhoids, the immediate postoperative course and final results are explained. A comparison of the Milligan-Morgan's and Parks' hemorrhoidectomy from different aspects, especially strictly defined indication, demonstrates that the Parks' operation shows better findings in both postoperative phase and final results. PMID- 7471957 TI - [Long-term experiences with sphincteroplasty using the Wreden-Stone method in the management of ano-rectal incontinence]. AB - After showing the reasons for anorectal incontinence and describing the different methods of surgical treatment, the sphincterplasty technique according to Wreden Stone is presented, as modified by the author in 1962: (1) a greater distance from the incision to the anus helps to reduce the risk of infection; (2) the strip of fascia lata winding around the anal canal is sutured to the musculus gluteus maximus on both sides. The functional results of the nine patients operated on by this method were subjectively much better than the objective results yielded by electro-myographic and manometric examinations; for only one patient were both kinds of results poor. Other methods are also discussed. At present the new treatment using smooth muscle from the large intestine is preferred, possibly combined with the sphincterplasty presented and additional use of striated muscle. PMID- 7471958 TI - [Experiences with the use of shoulder joint endoprosthesis]. AB - Total shoulder replacement has been successful as treatment for patients with irreparably damaged and painful glenohumeral joints, though improvement of joint motion is limited. Our experiences with 101 alloplastic operations on the human shoulder are reported for further design and construction of total shoulder joint prostheses. The type of prosthesis chosen depends on the degree of joint damage. A dorsal approach is recommended, carefully preserving or restoring function of the musculo-tendinous cuff mechanism. PMID- 7471960 TI - [Labor laws for assistant and head physicians]. PMID- 7471959 TI - [Patient education. Study of the interaction with surgical patients]. PMID- 7471961 TI - [Operative therapy of bone metastases]. PMID- 7471962 TI - [Surgery of lung metastasis]. PMID- 7471964 TI - [Value of circular mechanical colon and rectum anastomosis. Prospective study of 91 patients]. AB - A prospective study was carried out on 91 patients to evaluate colonic and rectal anastomoses constructed with an EEA stapling gun. The concept comprises the following: orthograde lavage, cecal fistula or colostomy, intraoperative sufficiency control of the anastomosis, intraoperative direct measurement of the height of the anastomosis, postoperative X-ray control of the anastomosis using standardized techniques, classification of the cases into three groups according to the height of the anastomosis: 3-5 cm, and over 9 cm. Characterization of the patients was accomplished biostatistically using the Median-Quartil system. Primary hospital letality of 2% was due to heart failure. Imparied wound healing occurred in 11%. Secondary suture dehiscence was roentgenologically identified in a total of 18% of which only 7% showed clinical symptoms. Stenosis was determined by X-ray in 11% with only 1% demonstrating a temporary clinical correlation. For especially low rectal anastomoses at 3-5 cm the EEA stapling gun appears to have a great technical advantage over hand-placed sutures, allowing extension of indications for low anastomoses. PMID- 7471963 TI - [Safe aboral distance in the sphincter-preserving resection of the rectum]. AB - The correlation between the aboral margin of clearance and the end results after anterior and deep anterior resection of the rectum were analyzed for 293 patients documentated in the Erlangen clinical-pathological registry of colorectal cancers. The distance between the aboral margin of the carcinoma and the resection line was measured under standardized conditions. The best prognosis (age-corrected 5-year survival rate and frequency of local recurrence) is given if the margin of clearance is more than 30 mm on resected fresh specimen without tension. This distance corresponds with 50 mm on the mobilized rectum in situ. This rule applied to carcinomas of Dukes' stage A and/or grade 1 as well as to more advanced tumors or tumors with a higher grade of malignancy. PMID- 7471965 TI - [Choice of operative technic in the treatment of liver diseases]. AB - Between 1974 and 1979 resection of the liver was carried out for malign or benign tumors in 42 patients. Lobectomy was performed in 17 and hemihepatectomy in 25 cases (11 times right-sided, 14 times left-sided). Hemihepatectomy with preliminary ligation of the vessels showed a lower rate of complication than the time-saving lobectomy. Mortality rate was 0% in left-sided hepatectomy, 12% in lobectomy, and 27% in right-sided hepatectomy. The cause of death was liver failure in three cases of hepatectomy because of preexisting damage of the remaining lobe showing no compensatory hypertrophy. In accordance with the literature, we conclude that right-sided hepatectomy can be performed only if hypertrophy of the left lobe is present. PMID- 7471967 TI - [Operative management of radius head fractures in children]. PMID- 7471966 TI - [Theoretical and clinical aspects of fibrin stabilization in the post-operative phase]. AB - The level of factor XIII circulating in the blood plasma was determined with a commercially available screening method of the Behringwerke, Marburg, Germany. In a randomized study, the blood of 76 of both sexes and different ages was analyzed before and after surgery. In most extensive laparotomies, factor XIII declined as an expression of the vascular processes during and after the operation. The same effect was demonstrated after surgical excision of stomach or of colon carcinomas. Concurrent with the factor XIII level, fibrinogen concentration was also reduced. While the replenishment of factor XIII extended over a period of several days postoperatively, the fibrinogen synthesis was increased and exceeded normal levels after 2 to 3 days. Inspite the operational activities, factor XIII was never so low that a substitution was indicated. PMID- 7471968 TI - [Definition of hepatic anatomical regions and their value during hepatectomy (author's transl)]. PMID- 7471969 TI - [Resection-anastomosis of the trachea for non-tumoural tracheal stenosis. 68 cases (author's transl)]. PMID- 7471970 TI - [Aberrant liver lobes, ectopic, and accessory livers. Two cases of small ectopic livers (author's transl)]. PMID- 7471971 TI - [Justification of the routine combination of partial mastectomy with axillary lymph node dissection followed by irradiation in the treatment of small carcinomas of the breast (435 cases, including 134 with more than 5 years follow up) (author's transl)]. PMID- 7471972 TI - [Double compression of the coeliac trunk and the superior mesenteric by the arcuate ligament (author's transl)]. PMID- 7471973 TI - [Pulmonary excisions and exacerbation of old peptic ulcer. Attempt at prevention by vagotomy-pyloroplasty (author's transl)]. PMID- 7471974 TI - [Leading principles for controlled hepatectomies (author's transl)]. PMID- 7471975 TI - [Complete haemostasis clamp for hysterectomy. New technique of hysterectomy for fibroids (author's transl)]. PMID- 7471976 TI - [Computer tomography examination of spinal fractures with nerve-root complications (author's transl)]. PMID- 7471977 TI - [Caroli's disease associated with intrahepatic lithiasis and gallbladder stones. Remission lasting more than three years after double drainage with seton (author's transl)]. PMID- 7471978 TI - [Angiofollicular lymphoid hyperplasia (Castleman's disease). First case of meso sigmoid localisation (author's transl)]. PMID- 7471979 TI - [Perineal approach for excision of villous tumors of the lower rectum and anal canal by anterior rectotomy after anal sphincter section (author's transl)]. PMID- 7471980 TI - [Gastrointestinal tumours of muscular origin. 11 cases (author's transl)]. PMID- 7471981 TI - [The treatment of peptic stenosis of the esophagus. 40 cases (author's transl)]. PMID- 7471982 TI - [Personal experience in the surgical correction of esophageal stenosis due to scarring (author's transl)]. PMID- 7471983 TI - [Esophagoplasty for caustic stenosis. Apropos of 21 cases]. PMID- 7471984 TI - [Mechanical anastomosis with the EEA stapler in esophageal surgery]. PMID- 7471985 TI - [Influence of organic disease in drivers on road accidents (author's transl)]. PMID- 7471987 TI - [Hydatid cysts of the lung. The problem of recurrence]. PMID- 7471986 TI - [Continent ileo-cecal bladder: 33 cases (author's transl)]. PMID- 7471988 TI - [Tomodensitometry in hydatid cysts]. PMID- 7471989 TI - [Leo Buerger's disease: a review and report on 23 cases (author's transl)]. PMID- 7471990 TI - [Current therapy of malpighian cancers of the anal canal: a report on 51 cases (author's transl)]. PMID- 7471991 TI - [High strictures of the thoracic esophagus and their surgical treatment (author's transl)]. PMID- 7471992 TI - [Peptic stenosis or esophageal carcinoma: difficulties of the pre-operative diagnostic and value of the surgical exploration (author's transl)]. PMID- 7471993 TI - [Repeat operations following surgery for abdominal cancers (author's transl)]. PMID- 7471994 TI - [Post-traumatic laryngo-tracheal disinsertion with fracture of the cricoid cartilage and tearing of the recurrent laryngeal nerves. Therapeutic considerations based upon an experience of 8 cases (author's transl)]. PMID- 7471995 TI - [Studies of the physiological variations of leukocyte in peripheral blood during pregnancy (author's transl)]. PMID- 7471996 TI - [Experimental studies of intra-amniotic pargyline and/or reserpine for termination of midterm pregnancy (author's transl)]. PMID- 7471997 TI - [Diagnosis and treatment of penetration of uterine wall by IUD (author's transl)]. PMID- 7471998 TI - [Experience in the treatment of large vesicovaginal fistula (author's transl)]. PMID- 7471999 TI - [Stress urinary incontinence in the female (analysis of 116 cases (author's transl)]. PMID- 7472000 TI - [Vaginoplasty (analysis of 52 cases) (author's transl)]. PMID- 7472001 TI - [Ureteral injuries in women (clinical analysis of 32 cases) (author's transl)]. PMID- 7472002 TI - [Ovarian cystic teratoma: clinico-pathological analysis of 547 cases (author's transl)]. PMID- 7472004 TI - [The diagnosis and management of uterine myoma complicated with pregnancy (analysis of 180 cases) (author's transl)]. PMID- 7472003 TI - [Yolk sac tumor of the ovary (a report of 6 cases) (author's transl)]. PMID- 7472005 TI - [The characteristics of ovarian serous tumors of borderline malignancy (author's transl)]. PMID- 7472006 TI - [Clinical observations on fetal heart rhythm (author's transl)]. PMID- 7472007 TI - [Occipitopubic position and occipitosacral position at pelvic inlet (clinical analysis of 45 cases) (author's transl)]. PMID- 7472008 TI - [The normal pelvis of Congo women (author's transl)]. PMID- 7472009 TI - [A brief review of the research work on toxemia of pregnancy in Shanghai (author's transl)]. PMID- 7472010 TI - [Clinical observation of amniotic fluid creatinine as an index of fetal maturity (author's transl)]. PMID- 7472011 TI - [Thoracic kyphosis complicating pregnancy---an analysis of 60 cases (author's transl)]. PMID- 7472013 TI - [Measurement of menstrual blood loss (author's transl)]. PMID- 7472012 TI - [Pulsogram in women: preliminary report of 1,137 cases (author's transl)]. PMID- 7472014 TI - [Serum hormonal levels during normal menstrual cycles and in menopausal women (author's transl)]. PMID- 7472015 TI - [The cytology of endometrial cavity with retained IUD (author's transl)]. PMID- 7472016 TI - [Diagnosis of intrauterine adhesion after uterine curettage---an analysis of 32 cases (author's transl)]. PMID- 7472017 TI - [Observation of change in serum purple colorimetric response during diagnosis and treatment of ovarian cancer (author's transl)]. PMID- 7472018 TI - [A clinico-pathological analysis of invasive carcinoma of the cervix (author's transl)]. PMID- 7472019 TI - [Carcinoma in situ of the uterine cervix---a clinical analysis of 166 cases (author's transl)]. PMID- 7472020 TI - [Preoperative radiation and surgical treatment of stage II carcinoma of uterine cervix (author's transl)]. PMID- 7472022 TI - Meiotic segregation of five different reciprocal translocations in the onion fly, Hylemya antiqua (Meigen). AB - For one translocation (T14) with short interstitial segments in Hylemya antiqua significant differences in segregation behaviour between males and females were observed. In males the ratio of alternate:adjacent 1:adjacent 2 was approximately 7:3:0 and in females about 8:1:3. This difference is attributed to the difference in type of chromosome association. Female meiosis is chiasmate and male meiosis is achiasmate. It is suggested that meiotic pairing in males results in relative short "Coorientation Determining Distances" (CDDs) between homologous centromeres which favours alternate and adjacent 1 segregation. In females because of non localized chiasmata on the average no differences in CDD between homologous and nonhomologous centromeres are expected. This might explain the occurrence of coorientation between non-homologous centromeres resulting in adjacent 2 segregations. Four other translocations with longer interstitial segments than T14 showed in males as well as females predominantly an alternate and adjacent 1 segregation, adjacent 2 was hardly found (0-3.6%). The longer distance between non-homologous centromeres is probably the reason. PMID- 7472021 TI - Localization and characterization of rDNA in Drosophila tumiditarsus. AB - Using in situ nucleic acid hybridizations, the genes that code for 28, 18 and 5S rRNA have been localized in the polytene chromosomes of Drosophila tumiditarsus. The 5S genes are found at a single site near the centromere of the second chromosome, whereas the 28 and 18S genes are found at the nucleolar organizer region of the dot chromosome. The dot chromosome has been previously described as alpha-heterochromatic. However, our cytochemical and autoradiographic results do not support such a conclusion. The autoradiographic results reveal that the dot chromosome is transcriptionally active and is not late-replicating, as is expected of alpha-heterochromatin. Further, the dot chromosomes possess none of the usual staining characteristics of heterochromatin except for its lack of polytene bands. Using rRNA-DNA filter hybridizations, we find that the rDNA of D. tumiditarsus salivary glands is under-replicated. This is the first species of Drosophila where the rDNA in not found on the sex chromosomes, and is the first report of an under-replicated autosomal locus which is not located in heterochromatic blocks. PMID- 7472023 TI - [Some special clinical manifestations of Behcet's disease---report of illustrative cases and review of literature (author's transl)]. PMID- 7472026 TI - [Clinical and immunological analyses in 25 cases of xerotic syndrome (author's transl)]. PMID- 7472024 TI - [Cardiac manifestations of systemic lupus erythematosus (author's transl)]. PMID- 7472025 TI - [Clinical analysis of 38 cases of lupus nephritis (author's transl)]. PMID- 7472027 TI - [Investigation of cellular immune state in patients with viral hepatitis-- observation on the delayed vaccine type skin hypersensitivity reaction (author's transl)]. PMID- 7472029 TI - [Chylus ascitis---report of 8 cases and discussion on its etiology (author's transl)]. PMID- 7472028 TI - [Clinical and pathological manifestations in 37 cases of catecholaminosis (author's transl)]. PMID- 7472030 TI - [A comment on analysis of fever types and drug reactions for differential diagnosis of unknown causes low grade fever (author's transl)]. PMID- 7472031 TI - [Alterations of ECG caused by bephenium (author's transl)]. PMID- 7472032 TI - [Treatment of schistosomiasis with furan-propylamine and dipterex--its reaction, effectiveness and evaluation (author's transl)]. PMID- 7472033 TI - [Discussion minutes: prevention and treatment of coronary heart disease, angina pectoris and arrhythmias with Chinese traditional and Western medicine (author's transl)]. PMID- 7472034 TI - [Immunology relevant to clinical medicine (author's transl)]. PMID- 7472035 TI - [A composite report on immunoblastic lymphadenopathy in 11 cases (author's transl)]. PMID- 7472036 TI - [Oral glucose tolerance test in 269 normal subjects aged over 40 and diagnostic criteria of diabetes mellitus (author's transl)]. PMID- 7472037 TI - [Recurrent polychondritis and its cyclophosphamide therapy (author's transl)]. PMID- 7472038 TI - [Diagnostic value of smooth tongue and fissure tongue in stomach diseases (author's transl)]. PMID- 7472040 TI - [Non-professional mercury poisoning (author's transl)]. PMID- 7472039 TI - [Measurement of P(A-a)O2 and its clinical significance (author's transl)]. PMID- 7472041 TI - [Clinical analysis of 20 cases of lymphosarcoma cell leukemia (author's transl)]. PMID- 7472042 TI - [Rosette formation by mice red cells and human lymphocytes (author's transl)]. PMID- 7472043 TI - [Clinical analysis of 21 cases of jaundice caused by nithiocyaminum (amoscanate, C9333 GO/CGP4540) (author's transl)]. PMID- 7472045 TI - [Comment: Some suggestions on the researches on prevention and treatment of diabetes mellitus (author's transl)]. PMID- 7472044 TI - [Correlation of immunology with clinical medicine (author's transl)]. PMID- 7472046 TI - [Complications of diabetes mellitus (author's transl)]. PMID- 7472048 TI - [Clinical significance of beat-to-beat variation in vegetative nervous system dysfunction in diabetes mellitus (author's transl)]. PMID- 7472047 TI - [Diabetes mellitus and coronary heart disease (author's transl)]. PMID- 7472049 TI - [Study on ecological characteristics of the chief malaria vector Anopheles balabacensis balabacensis in Hainan Island and the effects of control experiment (author's transl)]. PMID- 7472050 TI - [Antitoxic response of paraoxon-hemocyanin immunized rabbits (author's transl)]. PMID- 7472051 TI - [Toxicological studies of inorganic fluoride in drinking water (author's transl)]. PMID- 7472052 TI - [Serological survey of human influenza antibodies in domestic animals, poultry and wild birds (author's transl)]. PMID- 7472053 TI - [New methods for measurement and comparison of survival rates of neoplasms and immunization results (author's transl)]. PMID- 7472054 TI - [Decompression diseases: a report of 240 cases (author's transl)]. PMID- 7472055 TI - [Principles for determining fluoride standard in drinking water (author's transl)]. PMID- 7472056 TI - [Radiological analysis of 15 cases of silicosis complicated with middle-lobe syndrome (author's transl)]. PMID- 7472057 TI - [Food-borne endemic fluorosis (author's transl)]. PMID- 7472058 TI - [Accidental mushroom poisoning (author's transl)]. PMID- 7472059 TI - [Pneumoconiosis in fireworks workers (author's transl)]. PMID- 7472060 TI - [A survey of pneumoconiosis sufferers in a mica mine (author's transl)]. PMID- 7472061 TI - [Raising the level of surgical treatment of lung cancer (author's transl)]. PMID- 7472062 TI - [Surgical treatment of atrioventricular canal malformations (author's transl)]. PMID- 7472063 TI - [Congenital coronary arterial fistula and its treatment (author's transl)]. PMID- 7472064 TI - [The clinical use of high flow rate extracorporeal circulation in 100 cases (author's transl)]. PMID- 7472065 TI - [Operative treatment of 37 cases of bronchial carcinoma aged 70 or more (author's transl)]. PMID- 7472066 TI - [Pathologic factors influencing survival after resection of bronchogenic carcinoma (author's transl)]. PMID- 7472067 TI - [Evaluation of Barrett technique in 180 cases of pulmonary hydatid cyst (author's transl)]. PMID- 7472068 TI - [Application of mathematical statistics in differential diagnosis of acute intestinal obstruction (author's transl)]. PMID- 7472069 TI - [Experiences in surgical treatment of intrahepatic acute massive biliary tract hemorrhage: report of 23 cases (author's transl)]. PMID- 7472070 TI - [Pancreatoduodenectomy in the treatment of periampullary carcinoma: report of 23 cases (author's transl)]. PMID- 7472071 TI - [Choledocholithiasis with septic shock treated with combined traditional Chinese and western medicine: report of 111 cases (author's transl)]. PMID- 7472072 TI - [Preliminary report on spleno-cava venous shunt (author's transl)]. PMID- 7472073 TI - [Therapeutic results of left hemihepatectomy for primary hepatic carcinoma (author's transl)]. PMID- 7472074 TI - [Experiences in the diagnosis and treatment of carotid body tumors: report of 14 cases (author's transl)]. PMID- 7472075 TI - [Treatment of traumatic internal carotid-cavernous sinus fistula with electrothrombosis: report of a case (author's transl)]. PMID- 7472076 TI - [Intracranial arteriovenous malformations in children (author's transl)]. PMID- 7472077 TI - [Preliminary experiences in the treatment of hemorrhoids with laser (author's transl)]. PMID- 7472078 TI - [Combined treatment with traditional Chinese and Western medicine for chronic prostatitis (author's transl)]. PMID- 7472079 TI - [Experiences in the reconstruction of total and subtotal eyelid defects (author's transl)]. PMID- 7472080 TI - [Aponeurosis palmaris and palmar fascia spaces (author's transl)]. PMID- 7472081 TI - [TJ-bone cement: Preparation and clinical application (author's transl)]. PMID- 7472082 TI - [Four measurements of proximal end of femur in 279 normal adults (author's transl)]. PMID- 7472083 TI - [Autotransplantation of thumb: a case report (author's transl)]. PMID- 7472084 TI - [Prosthetic replacement of cervical vertebrae: result of long-term follow-up (author's transl)]. PMID- 7472085 TI - [Maffucci's syndrome: report of 2 cases (author's transl)]. PMID- 7472086 TI - [Some principles concerning application of compression plate (author's transl)]. PMID- 7472087 TI - [Clinical pathological conference: Jaundice, pancreatic atrophy, jaundice again after operation, sudden death (author's transl)]. PMID- 7472088 TI - [On how to perform clinical researches (author's transl)]. PMID- 7472089 TI - Role of prolactin and the renin-angiotensin system in mediating dopaminergic control of aldosterone secretion in the rat. AB - Blockade of dopaminergic pathways increases aldosterone levels by mechanisms that are not well delineated. Since both prolactin (PRL) and plasma renin activity (PRA) also increase after administration of dopaminergic antagonists, the aldosterone increments may be secondary to these changes. To address these questions, the relationship between plasma aldosterone (PA) and PRL responses to 2 different dopamine receptor antagonists, haloperidol and metoclopramide (MCP) was examined in rats. The PA response to MCP was compared before and after blockade of the renin-angiotensin system with saralasin and after pre administration of L-dopa. MCP administration produced significant and parallel increments in PA and PRL whereas haloperidol increased PRL without any change in PA or PRA. L-dopa pre-treatment suppressed the early PA response to MCP. Hypophysectomy prior to MCP administration eliminated the PRL response but did not significantly alter the PA response to MCP. Our findings suggest that dopamine has an inhibitory action on the adrenal gland production of aldosterone acting independently of changes in PRL and the renin-angiotensin system. PMID- 7472091 TI - What is the link between vascular smooth muscle sodium pumps and hypertension? PMID- 7472090 TI - Accumulation of norepinephrine by tissue slices from spontaneously hypertensive rats. AB - Accumulation of 3H-1-norepinephrine (3H-1-NE) by heart and spleen slices from different aged spontaneously hypertensive rats (SHR) was compared with accumulation by slices from comparably aged normotensive rats (Wistar-Kyoto, WKY). Between weeks 4 and 9 there was a significant increase in accumulation by heart slices of the WKY; there was no further increase at weeks 13 and 16. The developmental pattern in slices of WKY spleen was different. Accumulation was the same at weeks 4 and 9; subsequently, there were significant increases at weeks 13 and 16. Slices of heart from SHR showed the same pattern of development as those from WKY. However, accumulation by slices of SHR spleen was significantly different from that by slices of WKY spleen. At all ages studied, SHR slices accumulated less 3H-1-NE. Furthermore, there was no significant increase in accumulation with age. The data show: 1. the course of development of the adrenergic neuron transport system is different in heart and spleen, and 2. accumulation of 3H-1-NE by SHR spleen, but not heart slices, is different from accumulation by the respective control tissues. PMID- 7472092 TI - Effects of diet on exaggerated natriuresis in hypertension. AB - The effects of high and low salt diet on exaggerated natriuresis after volume expansion were examined in two groups of hypertensive patients, 6 with labile and 6 with fixed uncomplicated essential hypertension. Fixed hypertensives eliminated the administered salt-load faster than the labile hypertensives, and diet had no effect on the exaggerated natriuresis of both groups. No association was observed between plasma renin activity and natriuresis in both groups of patients. We conclude that, 1) diet did not affect the saline-induced natriuresis in hypertensive patients, 2) there was no apparent association between plasma renin activity and exaggerated natriuresis in hypertension, 3) other factors such as the capacitance system and vasopressin may play an important role in volume expansion natriuresis, especially in fixed hypertensives. PMID- 7472093 TI - Abnormal saphenous vein composition in human hypertension. PMID- 7472094 TI - Control of hindlimb vascular resistance and vascular responsiveness in DOCA-salt hypertensive dogs. PMID- 7472095 TI - Short flexible fiberoptic colonoscopy in routine office examinations. AB - Through experience with 1,464 examinations using the short flexible fiberoptic colonscope, the author concludes it to be the preferred instrument for endoscopy of the distal bowel. He finds it superior to the rigid instrument in detection of polyps and early malignant disease of the distal descending and sigmoid colons. The flexible instrument is generally less uncomfortable for the patient and is readily accepted as a routine part of a thorough colonic and rectal examination. PMID- 7472096 TI - A rectal sump in treatment of severe diarrhea. PMID- 7472097 TI - Anal complications in Crohn's disease. AB - Anal fissures, fistulas, and abscesses occurred as complications in 22 per cent of our population of 1,098 patients with Crohn's disease. Crohn's colitis was much more frequently associated with an anal lesion than Crohn's disease of the small bowel (52 per cent vs. 14 per cent). When an anal lesion is the manifesting sign, Crohn's disease will soon develop elsewhere in the intestine. Since these lesions frequently herald the onset of intestinal Crohn's disease, the physician must always be aware of the possibility of inflammatory bowel disease when dealing with suspicious anal lesions. PMID- 7472098 TI - Colorectal carcinoma in patients less than 40 years of age: pathology and prognosis. PMID- 7472099 TI - Polyps in a low-risk colonic cancer population in Colombia, South America. AB - Colonic polyps were systematically searched for in 508 specimens collected from unselected autopsies in individuals more than 10 years of age from Medellin, Colombia, a low-incidence geographic region for colorectal cancer. Adenomatous polyps have a low prevalence rate in both sexes, comparable to that found in Cali, Colombia and other populations with low risk for cancer. They were found more frequently in the right side of the colon and are rare in the sigmoid colon. They begin to appear in the third decade of life in both sexes. Prevalence rates for single and multiple polyps increase with age in both sexes, males showing higher rates than females at every age group. Adenomatous polyps in this population are usually less than 3 mm in diameter and do not show villous changes or epithelial atypia. Hyperplastic polyps were more frequently observed than adenomatous polyps and rates show a progressive increase with age in both sexes. Multiplicity also increases with age. Most polyps are located in the low rectum and sigmoid followed by the cecum and ascending colon. Usually hyperplastic polyps are smaller than 3 mm in diameter and do not show epithelial atypia. Juvenile polyps were infrequently found in this series because children less than 10 years old were excluded. Diverticular disease was also rare and, when found, was not associated with adenomatous polyps. The association between adenomatous polyps and colonic cancer is discussed based on findings in this report. PMID- 7472100 TI - Mucosal enteritis: a complication of the continent ileostomy. AB - Mucosal enteritis, a nonspecific inflammation of both the mucosa and submucosa, is a complication seen in 13 per cent to 43 per cent of patients with a continent ileostomy. Six cases are presented with emphasis on history, diagnosis, and treatment. Because the inflammation is secondary to an overgrowth of anaerobic bacteria in a functional blind loop, treatment consists of drainage and antibiotics. Metronidazole may be the drug of choice. Refractory cases will benefit only by resection and conversion to a Brooke ileostomy. PMID- 7472101 TI - Bile duct carcinoma associated with chronic ulcerative colitis. AB - Patients with chronic ulcerative colitis are prone to a variety of liver disorders. This case report illustrates development of bile duct carcinoma in a patient with long-standing inactive colitis. The report emphasizes the association of chronic ulcerative colitis with bile duct carcinoma and discusses the radiologic preoperative evaluation of the ulcerative colitis patient who develops jaundice. PMID- 7472102 TI - Small-bowel obstruction secondary to pericecal hernia. AB - Pericecal hernia is a rare type of internal hernia and, though congenital in origin, may occur at any age, usually with symptoms of acute intestinal obstruction. The pathophysiology and character of these defects are described, and the management of this situation in an 80-year-old patient is reported. Prompt operative intervention to avoid the associated high mortality secondary to strangulation is urged. It is suggested that closure of these defects be performed if incidentally found at laparotomy. PMID- 7472103 TI - Solitary perforated diverticulum of the ascending colon: report of two cases. AB - Diverticulosis of the ascending colon is rare. Perforation of a diverticulum is very uncommon and at times it is difficult to differentiate from carcinoma. Two unusual cases of perforated solitary diverticula of the ascending colon are reported, and literature is briefly reviewed. PMID- 7472104 TI - Angiography with epinephrine and beta receptor blocker in diagnosis of sigmoid lesions. AB - Various sigmoid lesions, seen on barium enema of 39 patients, were investigated with angiography of the inferior mesenteric artery using three series. The first was performed with contrast medium only, the second after use of epinephrine, and the third after a beta-blocking agent (propranolol) combined with epinephrine. The patients were followed for more than three years. Eleven carcinomas were detected, all diagnosed angiographically. Carcinoma, radiation fibrosis, and diverticulitis with fibrosis may appear similar on pharmacoangiography, but the inflammatory reaction seen, for example in diverticulitis, is different. These findings are important because they can have an influence on the indications and selection of the most suitable operation to be performed. PMID- 7472105 TI - The comparative electrophysiologic and hemodynamic effects of verapamil in puppies and adult dogs. AB - Verapamil has been studied extensively in adult animals. However, there is limited data about the effects of this antiarrhythmic agent in young animals. We studied the electrophysiologic and hemodynamic effects of verapamil in 5 awake, chronically instrumented puppies and in 6 anesthetized, acutely instrumented puppies. We compared the responses in puppies to those in 7 awake, chronically instrumented adult dogs and in 6 anesthetized, acutely instrumented adult dogs. In the awake, chronically instrumented animals, we measured heart rate and the effective refractory period of the atrio-ventricular specialized conduction system (ERP-AVSCS). In the anesthetized, acutely instrumented preparation, we measured heart rate, ERP-AVSCS, mean right atrial and systemic arterial blood pressure, and velocity of blood flow in the aorta, carotid and femoral arteries. We infused verapamil intravenously at doses of 50, 100, 200 and 400 microgram/kg. Verapamil (400 micrograms/kg) increased the ERP-AVSCS by 64% in awake, chronically instrumented puppies; 62% in awake, chronically instrumented adult dogs; 125% in anesthetized, acutely instrumented puppies, and 120% in anesthetized, acutely instrumented adult dogs. The effect of verapamil upon heart rate and ERP-AVSCS was more pronounced in anesthetized animals than in awake animals. Mean systemic arterial blood pressure was reduced by verapamil in anesthetized, acutely instrumented puppies and adult dogs. Verapamil reduced the velocity of aortic, femoral and carotid artery blood flow in anesthetized, acutely instrumented puppies. PMID- 7472106 TI - Prevention of umbilical artery catheter clots with heparinized infusates. AB - 49 neonates requiring umbilical artery catheters (UACs) were randomly assigned to receive standard or heparin-containing infusates. 3 of 23 (13%) of the patients receiving heparin had catheters removed because they became functionally occluded compared to 15 of 26 (58%) in the control group (p less than 0.005). 4 of 13 (31%) single injection aortograms obtained in control infants demonstrated thrombi, compared to none of 7 in the heparin group. 1 patient in the heparin group had an aortic clot demonstrated at post-mortum examination. There were neither clinical coagulopathies nor abnormalities of partial thromboplastin time attributable to the administration of heparinized fluids. Heparinization of UAC infusates appears to be a safe method of reducing the risk of catheter occlusion. Heparin effect on large vessel clot risk remains unproven. PMID- 7472107 TI - Age-dependent variations in the distribution of methaqualone in rat. AB - Rats at 1, 4, 8, 12, 20 and 60 postnatal days received orally 5 mg/kg 14C methaqualone (MTQ) and were sacrificed at 0.5, 1.5 and 6 h. Several tissues were analyzed for unchanged MTQ. In most cases, concentrations reached peak levels at 1.5 h. Highest concentrations were found in the liver followed by the lung, spleen, heart, kidney, eye, and brain. Levels of MTQ were higher in all organs of 1- to 12-day-old rats at all time intervals than the corresponding organs of 20- and 60-day-old rats, apparently resulting from initial higher accumulation and slower rate of disappearance. The 1.5-hour brain:plasma concentration ratio in 8 day-old rats was 0.82 and progressively declined to 0.27 in 60-day-old rats; a similar trend with respect to changes in concentration ratio also occurred for the eye, spleen, kidney, and heart. The fact that the age-related reduction in tissue:plasma concentration ratios occurred in several tissues including brain suggests generalized decreases in tissue accumulation of MTQ. The reduction in brain:plasma concentration ratios, therefore, cannot be ascribed to the blood brain barrier which develops progressively with the advancing age. PMID- 7472108 TI - Effect of ascorbic acid (vitamin C) on the epiphyseal plate of young guinea pigs receiving prednisolone. AB - The effect of excess ascorbic acid on the epiphyseal plate of young guinea pigs receiving prednisolone over a period of 28 days has been investigated. The results show that the structural and morphologic changes caused in the epiphyseal plate of the animals receiving prednisolone are prevented by the simultaneous administration of excess ascorbic acid. These data are consistent with the hypothesis that pharmacological doses of ascorbic acid given concurrently to children receiving prolonged treatment with corticosteroids may prevent suppression of growth in these children. PMID- 7472109 TI - Ascorbic acid treatment in nephropathic cystinosis in identical twins. AB - Metabolic studies were performed on two HLA identical 9 1/2-year-old twins with nephropathic cystinosis during 14 months' participation in a double-blind study to evaluate ascorbic acid treatment for cystinosis. Replacement therapy was identical in both children throughout the study except for the use of ascorbic acid. Leukocyte cystine content was markedly abnormal and elevated in both children prior to, during, and after the ascorbic acid study. We postulate that the lack of improvement in growth or chemical parameters in the twin treated with ascorbic acid was due to the ineffectiveness of ascorbic acid in decreasing intracellular cystine content. PMID- 7472110 TI - The comparative hemodynamic effects of isoproterenol in chronically instrumented puppies and adult dogs. AB - Although isoproterenol is the sympathomimetic amine used most often to augment cardiovascular function in infants and children, little is known about the hemodynamic effects of this drug on the immature cardiovascular system. To compare the hemodynamic effect of isoproterenol in the young animal with that in the mature animal, we infused the drug into 11 puppies of 12-38 days of age, 12 puppies of 56-89 days of age and 10 adult beagle dogs. The animals were chronically instrumented and nonsedated during the study. Isoproterenol was infused in doses of 0.05, 0.125, 0.25 and 0.5 microgram/kg/min. Isoproterenol decreased systemic arterial mean blood pressure, increased heart rate and had no effect on renal blood flow in all three groups of animals. While cardiac output was increased and systemic vascular resistance decreased in the adult dogs, neither changed in puppies. Stroke volume increased slightly in adult dogs, but decreased in puppies. This refractoriness to change of cardiac output in immature dogs may be a function of elevated resting cardiac output, developmental changes of contractility or ventricular compliance or developmental changes of vascular smooth muscle responsiveness. PMID- 7472111 TI - Maturity-onset diabetes of the young: an illustrative case for control of diabetes and hormonal normalization with dietary management. AB - The efficacy of diet therapy used for one patient with the newly classified maturity-onset diabetes of the young (MODY) is reported. Using the conventional American Diabetes Association's food exchange system with appropriate caloric restriction, the patient's weight was reduced from 25% to 6% above ideal body weight in the 14-wk study. Results of three meal tolerance tests compared with a group of lean, normal subjects revealed a steady improvement from baseline and postprandial hyperglycemia, with concomitant normalization of the counterregulatory hormones as well as hemoglobin A1, free fatty acids, and subsequent euglycemia. This case demonstrates the effectiveness of dietary manipulation as the major therapy in MODY with obesity. PMID- 7472112 TI - Nucleated red blood cells and artifactual hypoglycemia. AB - A patient with chronic hemolytic anemia presented with an acute hemolytic crisis, a high count of nucleated red blood cells (NRBC), and artifactual hypoglycemia. Temporal events showed a parallel relationship between an increased number of nucleated RBCs and an excess in vitro consumption of glucose. PMID- 7472113 TI - The problem of "cheating" in the diabetic child and adolescent. AB - The frequency, causes, and consequences of "cheating" in diabetic children and adolescents were studied during summer camp. A philosophical approach is proposed for its understanding and management. PMID- 7472114 TI - Classification of diabetes. PMID- 7472115 TI - Relationships between control and serum lipids in juvenile-onset diabetes. AB - The relationship between control and serum lipids was examined in 147 children with juvenile-onset diabetes (JOD). Control was assessed by fasting blood glucose, 24-h urine glucose, and fast hemoglobin. There was a significant positive correlation (P less than 0.001) between each of the measures of control. Serum cholesterol (P less than 0.001), triglycerides (P less than 0.001), and low density lipoprotein plus very low-density lipoprotein cholesterol (P less than 0.001) were strongly positively correlated with control for the whole group of patients. High-density lipoprotein was not significantly correlated with control when the data for the whole group were analyzed but was significantly positively correlated with fasting blood glucose when patients were analyzed individually. PMID- 7472116 TI - Nutrition therapy in non-insulin-dependent diabetes mellitus. AB - The purpose of this study was to evaluate nutritional therapy for non-insulin dependent diabetic subjects by examining the changes in, and interrelationship of, the outcome criteria of efficacy, namely, blood glucose, body weight and serum lipids, total cholesterol, and triglycerides. Eight-two patients, who represented all those seen by one nutritionist at a diabetic education center, were contacted for long-term follow-up data, which was obtained on 80.5% of the patients. They had short-term follow-up 66.0 +/- 4.5 days after the initial visit and long-term follow-up 31.0 +/- 1.1 mo later. Four treatment groups were defined: group 1, diet only at entry and follow-up; group 2, oral hypoglycemic agents at entry and follow-up; group 3, intensified treatment (diet or oral agents at entry and oral agents or insulin at follow-up; and group 4, decrease in intensity of treatment (oral agents at entry but not at long-term follow-up). Serum glucose fell significantly (P less than 0.05) at short- and long-term follow-up in groups 1, 2, and 3. Body weight reductions were significant at short term follow-up in all groups and were significant at long-term follow-up only in group 1. Serum cholesterol was significantly reduced at short-term follow-up in group 2. Serum triglyceride changes were significantly reduced at short-term follow-up in groups 1 and 2 and at long-term follow-up in group 1. Serum glucose changes correlated with changes in serum cholesterol and triglycerides but did not correlate with body weight changes. These findings demonstrate the effectiveness of nutritional counseling, show that lowering serum glucose did not correlate with weight reduction, and suggest that more attention should be given to factors reducing serum glucose and less attention to weight loss in the treatment of non-insulin-dependent diabetes. PMID- 7472117 TI - Nutrient intake in a partly westernized isolated Polynesian population: the Funafuti survey. AB - Dietary data were obtained from 110 adult Polynesians (indigenous Funafutians) during a diabetes epidemiologic survey in Tuvalu. Funafuti (the main island of Tuvalu) was chosen as representative of partly urbanized population that has seen progressive socioeconomic changes in the previous 30 yr. The distribution of calories in the Funafuti diet is almost identical to that reported in the urbanized Micronesian population of Nauru and in Caucasian populations. Prevalence of diabetes was found to be five times higher than that reported in caucasian populations. Similarly, the prevalence of hypertension was 11% (equal to Caucasian figures), whereas hypertension is virtually unknown in traditional living populations. Prevalence of diabetes, and degree of obesity, is significantly greater in women than in men. It is possible that this Polynesian group may have a genetic susceptibility to diabetes, which may have been unmasked by change from a traditional to western life-style. Marked obesity is now a characteristic of many Polynesian and Micronesian populations, and must rank as a major causative factor for the diabetes and hypertension explosion. PMID- 7472119 TI - [Lipid balance and good cholesterol]. PMID- 7472118 TI - Patient recall of self-care recommendations in diabetes. AB - Twenty-four insulin-dependent diabetic patients were interviewed immediately after a follow-up visit to an outpatient pediatric clinic to determine which of the recorded instructions delivered by professionals were recalled by patients. The health care team reported giving an average of seven recommendations per patient, or a total of 168 items listed by team members as important. Patients recalled an average of two recommendations, or a total of 50 items, 40% of which had not been recorded by team members. Recommendations concerning diet, insulin dosage and injections, urine testing, and exercise represented 80% of those recalled by patients and only 58% of those recorded by professionals. The results were interpreted to suggest that health care teams focus on fewer items to insure communication of the most appropriate recommendations for individual patients. PMID- 7472120 TI - [Chronic sub-dural hematoma: 87 observations]. PMID- 7472121 TI - [Poland's syndrome: one observation]. PMID- 7472122 TI - [Esophageal candidiasis in Dakar: a case with pseudo-tumorous form]. PMID- 7472123 TI - [Clinical examination of a patient with vertigo]. PMID- 7472124 TI - [Epidemiological and serological assessment of bilharziasis of students at a school in southern Togo]. PMID- 7472126 TI - [Histological aspects of nephrotic syndrome in adult Black Africans in the internal medicine service at Dakar (33 cases)]. PMID- 7472125 TI - [The place of nephropathies among the causes of secondary arterial hypertension in Senegal]. PMID- 7472127 TI - [Myoglobinuria and renal insufficiency in the course of tropical myositis]. PMID- 7472128 TI - [Acute renal insufficiency after retro-placental hematoma]. PMID- 7472130 TI - Adaptive programs for analysis of the visual field by automatic perimetry--basic problems and solutions. Efforts oriented towards the realisation of the generalised spatially adaptive Octopus program SAPRO. PMID- 7472129 TI - The Manschot--van Peperzeel concept of the growth and metastasis of uveal melanomas. PMID- 7472131 TI - Dynamic range and stray light. An estimate of the falsifying effects of stray light in perimetry. AB - The role of intraocular stray light upon contrast threshold of two skilled human observers was studied by perimetric methods using the automatic perimeter Octopus. Stray light falsifies the contrast sensitivity profile of the blind spot when a critical test stimulus luminance, which differs for various targets, is exceeded. At still higher test stimulus luminance, which differs for various targets, is exceeded. At still higher luminance levels, because stray light effects increase, the blind spot shrinks and finally disappears. A series of high resolution measurements of the blind spot with the automatic perimeter Octopus provide a quantitative answer concerning the commencement and amount of this disturbance as a function of target size and target luminance. The amount of stray light, when using targets varying from 0 to 5 (Goldmann standard) is related to luminous power (target luminance x target area). Using first order assumptions about the stray light emitting characteristics of the disc and empirical data, one may conclude that an increase in luminance of target 0 from 10(3) to 10(5) asb only increases the effective dynamic range by about 2 dB (= 0.2 log units 3, the standard target as compared with target) size used in Octopus perimetry, at a luminance level of 10(3) asb. Falsification of sensitivity gradients and underestimates of depths of scotomata due to stray light effects may be an ever present danger in perimetric determinations. The useful dynamic range in perimetry appears to be limited by photon noise and noise in the neurovisual system on the one hand and by stray light interference on the other. PMID- 7472132 TI - Different functional changes recorded in open angle glaucoma and anterior ischemic optic neuropathy. AB - The retinal ganglion cell exhibits two types of functional change. The difference in response appears to depend on whether or not the retrobulbar portion of the optic nerve is actively involved. This may imply differences in the myelinated and non-myelinated portions of the optic nerve and associated structures. In open angle glaucoma, alterations in the sustained- and transient-like functions but not in the flashing repeat static test are found. That is, there are changes in spatial neural interactions, but there is no evidence of a visual fatigue or saturation-like effect. In optic nerve radiation damage without evidence of retinal vascular changes, in optic neuritis secondary to multiple sclerosis, chiasmal lesions, and ischemic optic neuropathy (considered here) varying degrees of visual fatigue or saturation-like effects are demonstrated with little or no change in the sustained- or transient-like functions. PMID- 7472133 TI - Proteins in tears from healthy and diseased eyes. AB - The levels of total protein in tears from healthy donors, conjunctivitis vernalis patients, and conjunctivitis follicularis patients, were 625, 1370 and 1160 mg% respectively. Serum albumin accounted for 3.3%, 43% and 67% of the total protein of tears from these groups, and the level of proteins probably synthesized by the lacrimal gland, was in tears from conjunctivitis follicularis patients only half the level in normal tears. By crossed immunoelectrophoresis with intermediate gel, 10 antigenic species could be recognized in normal tears, and of these the following were identified: Lysozyme, IgA, lactoferrin and serum albumin. In tears from patients with conjunctivitis vernalis three more immunoprecipitates were observed, of which one was due to IgG. No lysozyme could be demonstrated in tears from a case of conjunctivitis sicca by immunoelectrophoresis. In tears from healthy donors the mean level of IgA was 20 mg%, of IgG 3 mg%, and IgM could not be demonstrated. Rabbit anti-tear immunoglobulin did not precipitate a standard of human IgM in double immunodiffusion. In cases of conjunctivitis vernalis and follicularis the mean levels were increased to 80 and 114 mg% IgG, and 11 and 14 mg% IgM, but IgA was increased only to 32 and 41 mg%. It is assumed that the level of IgA in normal tears is almost entirely due to local synthesis, while serum albumin and other immunoglobulins may have escaped from the circulation by molecular sieving. The increased levels of immunoglobulins in inflammatory diseases is probably due to transudation. However, in blepharoconjunctivitis patients several tear samples with a high IgM and a low or zero level of IgG could be demonstrated. Possible explanations for this phenomenon are discussed. PMID- 7472134 TI - Histology of small melanomas. AB - In a series of 193 choroidal melanomas we collected the histological data of 25 small and 31 medium-sized tumours enucleated on the basis of the results of the 32P-test. Thirty four (34%) percent showed epitheloid cells and 70% showed scleral involvement, both percentages being higher than in most reports. We do not advocate observation and delay of treatment in case of small melanomas until growth is established clinically. PMID- 7472135 TI - Echography of intraocular tumours. PMID- 7472136 TI - Pseudoregression in a case of choroidal melanotic tumor. PMID- 7472137 TI - Photocoagulation of choroidal melanomas. PMID- 7472138 TI - The place of radiotherapy in the treatment of uveal melanoma. PMID- 7472139 TI - Radiation therapy: before or after enucleation. PMID- 7472140 TI - The natural course of untreated uveal melanomas. PMID- 7472141 TI - The natural history of uveal melanomas and its therapeutic consequences. PMID- 7472142 TI - Phosphorus uptake test in choroidal melanomas, naevi and haemangiomas. PMID- 7472143 TI - Prediction of abnormal umbilical cord position and intrapartum cord problems from the nonstress test. AB - A prospective analysis of the nonstress test revealed a group of patients who demonstrate typical variable decelerations associated with fetal movement. Such variable decelerations were observed only in patients who had the test performed for suspected intrauterine growth retardation or for postdatism. Almost half of the suspected small-for-gestational-age fetuses and fetuses of a postdate gestation who demonstrated variable deceleration at nonstress testing also demonstrated an umbilical cord in an abnormal position at delivery. The position of the cord in utero of the remaining patients with variable deceleration at nonstress testing was not determined; however, 88% of this group demonstrated significant variable deceleration during labor. An abnormally positioned cord was noted in half of the patients undergoing cesarean section. It appears that patients with small-for-gestational-age fetuses and those with postdate gestation are at higher risk of intrapartum cord compression problems when variable decelerations are noted with fetal movements during a nonstress test. PMID- 7472144 TI - Malignant melanoma of the vulva. AB - Malignant melanoma of the vulva is reviewed and four cases are presented. Fewer than 400 cases have been reported, from 0.05 to 0.5% of female genital cancers. Radical vulvectomy with inguinal lymph node dissection is of uncertain value and the prognosis usually is poor. Wide local excision alone is suggested for level II melanomas, and in such instances the prognosis may be favorable. PMID- 7472145 TI - Gastrointestinal sediment layering. A useful sign for the prenatal sonographic diagnosis of fetal intestinal obstruction. AB - An instance of fetal intestinal obstruction uncovered during serial sonography in a pregnancy complicated by previous jejunoileal bypass is discussed. The presence of abnormal cystic structures with a gravity-dependent sediment layering effect within the fetal abdomen provided a useful, and possibly specific, sign in the diagnosis of fetal gastrointestinal obstruction. PMID- 7472146 TI - Focal pregnancy-like change of the breast. AB - Columnar secretory cells similar to those of lactating breast are occasionally seen in otherwise quiescent breast lobules from nonpregnant patients. These foci often contain vacuolated, enlarged, pleomorphic, and hobnail cells that may be confused with neoplastic cells. Nuclear features of malignancy are lacking, however. We present a 47-year-old woman who had focal pregnancy-like change near an excised fibroadenoma and review the literature concerning this finding. PMID- 7472148 TI - Frozen section examination of endometrial curettings. An aid to the rapid diagnosis of ectopic pregnancy. AB - The maternal mortality resulting from extrauterine pregnancy has been related to delayed intervention following the onset of symptoms. Therefore, methods that can rapidly include or exclude an ectopic pregnancy are of value. In selected cases dilatation and curettage, with frozen section examination of the curetted tissue, has been utilized to confirm an abnormal intrauterine pregnancy and vitually rule out an extrauterine pregnancy. PMID- 7472147 TI - Lymphography and CEA in the diagnosis of metastases in patients with stage IB cancer of the cervix. AB - Fifty-nine patients with Stage IB cancer of the cervix underwent lymphography and were screened for carcinoembryonic antigen (CEA) levels before radical hysterectomy with pelvic lymph node dissection. Very strict criteria were used in the evaluation of the lymphograms in order to avoid false positive results. CEA levels were determined in plasma with a modified CEA-Roche kit, and values over 5 micrograms/liter considered as positive. Lymphography: Approximately 20% of the pelvic lymph node metastases could be detected in films taken 24 hours after the injection of the contrast medium. In patients subjected to combined treatment with intracavitary radium followed by operation, control lymphograms were taken preoperatively, 6 weeks after the radium insertions. In these films the metastases detection rate was 55%, with no false positive lymphograms. CEA: Levels over 5 micrograms/liter were found in 13 patients. Pelvic lymph node metastases were found in 10 of them, giving a metastases detection rate of 42%. One patient with lung metastases and one patient with metastases in the ovaries also has elevated levels of CEA. There was one patient with "false positive" elevation of CEA. The combination of CEA determinations and lymphography 6 weeks after the radium treatment yielded a total pelvic lymph node metastasis rate of 85%. PMID- 7472149 TI - The course and outcome of drug addiction. A follow-up study of 178 cases in Madurai, South India. AB - One hundred and seventy-eight subjects diagnosed as dependent on various agents and first seen between 1970 and 1974 were followed up for 6 - 10 years and data collected in respect of the persistence of drug-taking behaviour, marital and occupational adjustment and mortality. Data were obtained 173 cases. Drug-taking persisted continuously in 87 and intermittently in 11 subjects and abstinence was achieved in 40 cases. The majority showed maladjustment in occupation and marital relations. There was a mortality of 17% (29 deaths) including 16 suicides. Twenty eight (17%) had made suicidal attempts. The observations are discussed from some Indian cultural points of view with reference to the follow-up characteristics of different types of addiction. PMID- 7472150 TI - The relative validity of a chronological drinking record. AB - The results of chronological and a daily drinking record are compared. The chronological record was completed by interviewers who probed for information on all drinking episodes during the last week. The daily record covered only the last day and was completed by the respondent without interviewer assistance. Because of the recency of the information, the daily record was expected to yield the more valid results, but this hypothesis was not supported by the findings. Total alcohol consumption reported by the chronological method was significantly higher because men, but not women, reported fewer drinking episodes with the daily record for the same period. Daily recording may have placed too heavy a burden on the most frequent drinkers, most of whom were men. PMID- 7472151 TI - Socio-familial characteristics of black male heroin addicts and their nonaddicted friends. AB - To determine whether family, peer, and school variables are associated with divergent heroin-use patterns or career outcomes, we interviewed 183 black male heroin addicts, experimenters, and non-heroin users from Chicago's south side. While subgroup differences on socio-familial variables generally were not statistically significant, we did observe a number of trends in the expected direction. For example, addicts were more likely than experimenters or nonusers to come from broken homes, to have friends who were involved in serious types of drug use and other illegal activities, and to drop out of high school. At the same time, the pervasiveness of some of our measures of "social disharmony" across the sample suggest that, at least in some inner-city black neighborhoods, these patterns are perhaps becoming the norm. PMID- 7472152 TI - Knowledge of drug names determines associations to drug names more than habits for the drugs. PMID- 7472153 TI - Marihuana use by pregnant women: neurobehavioral effects in neonates. AB - In a prospective study of 291 mothers-to-be, 20 per cent reported some use of marihuana before and/or during pregnancy. A significant increase in symptoms associated with nervous system abnormalities was observed among babies born to heavy regular marihuana users and visuals responses were affected in two- to three-day-old infants in a dose-related fashion. No relationship between marihuana use and maternal weight again, length of gestation, duration of labor or birthweight was found. PMID- 7472154 TI - Alcohol consumption and rates of alcoholism in Greece. AB - It appears that the incidence of alcoholism is relatively low in Greece, in spite of high consumption of alcohol and absence of restrictive measures for its use. This may be related to the traditional integration of alcohol use in the social ritualistic structures and to family and social support of susceptible individuals in the small Greek communities. The differences between urban and rural areas support these assumptions but should perhaps be taken as a danger signal. In order to establish effective measures of prevention, epidemiological studies are needed to confirm the trends and identify the risk factors. PMID- 7472155 TI - What can radiotherapy achieve now? PMID- 7472156 TI - [The value of influenza vaccination]. PMID- 7472157 TI - [Intravenous streptokinase infusions in acute myocardial infarction. Restoring patency of occluded thrombotic coronary vessels (author's transl)]. AB - Intravenous treatment over 30 minutes with 500 000 U streptokinase was given to 21 patients with acute myocardial infarction. Serial measurements of serum CK-MB activity made recanalisation of the infarcted artery likely in 3 patients. Selective coronarography was performed in 18 patients during the acute infarction phase. Angiographic follow-up investigations were done 24 hours and after 3 weeks. In 3 cases only subtotal occlusion of the infarcted coronary artery was seen during the acute phase. A thrombus demonstrated in two patients could be lysed. In 8 out of 15 patients with complete coronary artery occlusion the infarcted vessel was reopened 20-60 minutes after onset of intravenous streptokinase infusion. In a further 4 patients recanalisation was observed 60 140 minutes after passing a guide wire or during an additional intracoronary infusion of streptokinase. The infarcted vessel remained occluded in 3 patients, one of whom died following myocardial rupture. In 12 patients with recanalisation of the infarcted artery during the acute phase a premature maximum of serial CK MB serum activity was seen after 13.5 +/- 2.9 hours in comparison with 20.9 +/- 5.3 hours in 122 patients treated conventionally. Serial enzyme determinations allow indirect conclusions as to a rapid recanalisation of an infarcted blood vessel. PMID- 7472159 TI - [Surrendering medical records and giving information to public assistance offices]. PMID- 7472158 TI - [Differential diagnosis of creatine kinase elevation: hypothyroid myopathy (author's transl)]. AB - An increase of creatine kinase (CK) activity in serum as a result of myopathy of skeletal muscle can point to hypothyroidism. In the differential diagnosis from myocardial infarction determination of CK isoenzymes is the most suitable test. In three women (aged 56, 70 and 72 years) with hypothyroid myopathy demonstrated in this manner, administration of thyroid hormone preparations normalised CK activity within 2-4 weeks. A little later thyroid hormone levels in serum were also normal, while histological changes in the musculature persisted rather longer. PMID- 7472160 TI - [Sonographic demonstration of the pancreatic duct]. PMID- 7472161 TI - [Diagnosis of early stomach cancers]. PMID- 7472162 TI - [Heparin-calcium and psoriasis vulgaris]. PMID- 7472163 TI - [Vaccination against hepatitis B]. PMID- 7472164 TI - [Halothane hazards for the team in the operation theatre (author's transl)]. AB - Reports on undesirable side effects of halothane in patients undergoing anaesthesia and in persons with long-term exposure in the operation theatre require establishment of safe upper limits and of methods for monitoring. The main metabolite trifluoroacetic acid is most suitable for biological monitoring. During systematic exposure with graded halothane concentrations (8, 24 and 50 ppm, 6 hours daily, 5 days weekly, for one and two weeks) trifluoroacetic acid concentrations in blood and urine were determined regularly. It was shown that for biologic monitoring estimation in blood is clearly preferential to estimation in urine because of a narrower scatter. At the end of one week of exposure a steady state concentration is found in blood. There is a linear relation between its height and the inhalation concentration of halothane. A trifluoroacetic acid concentration of 2.5 micrograms/ml is considered as a risk-free value. This corresponds to a maximal theatre air concentration of 5 ppm halothane. PMID- 7472165 TI - [Conservative primary operation of carcinoma of the breast: subcutaneous mastectomy, lymphadenectomy and radiotherapy (author's transl)]. AB - So-called nipple-preserving operations were performed as primary treatment of mammary carcinoma, predominantly in stage T1, in 135 women. The operation consisted of subcutaneous mastectomy, axillary lymphadenectomy, nipple biopsy and X-ray therapy. Most of the patients in whom more than two lymph nodes had carcinomatous changes also received chemotherapy. Survival of more than five years was achieved in 48 patients in stage T1. One woman died of carcinoma; in six there was further growth with metastases. In no case was there further growth in the nipple region. Although the numbers are small one can already state that the results are no worse than after more radical operations. PMID- 7472166 TI - [The physician's duty for documentation and right of the patient to look at the medical record]. PMID- 7472167 TI - [Cell therapy: expensive placebo or dangerous natural treatment procedure?]. PMID- 7472169 TI - [Life expectancy and size of the liver in the presence of liver metastases: study of the diagnostic significance of palpatory findings (author's transl)]. AB - A retrospective analysis of survival time of 348 patients with liver metastases indicates that both the extent of liver enlargement and also its increased firmness and changes at its surface are easily obtainable findings of prognostic value. In the presence of liver metastases there is a significant correlation between median life expectancy and size of the liver. PMID- 7472168 TI - [Autotransplantation of cryopreserved human parathyroid tissue (author's transl)]. AB - Following the third operation, aparathyroidism occurred in a 70-year-old female with primary hyperparathyroidism due to parathyroid hyperplasia. Part of the parathyroid obtained during the last operation was cryopreserved immediately after resection and replanted successfully after 15 days into the lower arm muscles. The function of the transplanted tissue was demonstrated by successful stepwise reduction of hormone substitution. Parathyroid hormone concentrations in the blood of the veins draining the transplanted tissue were more than double in comparison with the contralateral side. Functional and morphological investigations of the defrosted tissue prior to transplantation showed vitality and intact morphology. PMID- 7472171 TI - [What is the role of total blood viscosity, erythrocyte deformation and erythrocyte aggregation?]. PMID- 7472170 TI - [Admissibility of physical immobilization of patients]. PMID- 7472174 TI - [Conservative therapy of stomach ulcer]. PMID- 7472172 TI - [Clinical importance of the estimation of thyrotropin displacing antibodies (author's transl)]. AB - In 69 hyperthyroid patients with diffuse uptake in the scintigram, thyrotropin displacing antibodies (TDA) were measured before, during and after thyrostatic treatment. In a further 42 patients antibody activity was investigated before and 6-12 weeks after radio-iodine treatment, and in 29 patients it was measured before and after subtotal thyroidectomy. Persistence of demonstrable TDA during and after the end of drug treatment was in all cases indicative of recurrence of hyperthyroidism. On the other hand, disappearance of TDA activity was not a guarantee for prolonged remission as five patients had a recurrence despite loss of TDA activity. In a further nine patients hyperthyroidism reappeared with recurrence of TDA after varyingly long periods of remission. Thyrostatic treatment should be performed until disappearance of antibodies. Prolonged demonstration of TDA should lead to surgical intervention. Besides the usefulness in establishing the diagnosis of immunogenic hyperthyroidism (Graves' disease) TDA estimation may be of some prognostic value in the use of thyrostatics. In contrast, after radio-iodine treatment or operation the functioning capacity of the remaining thyroid tissue seems to be more relevant for the success of treatment than the possibly longer acting antibodies. PMID- 7472173 TI - [Phenobarbital in febrile convulsions of children (author's transl)]. AB - Phenobarbital (Luminal) was given to 41 children (aged 11 months to 4 1/2 years) with uncomplicated febrile convulsions. The drug was administered orally or intramuscularly at an initial dose of 10 mg/kg or 15 mg/kg, respectively. Twelve hours later and then every 24 hours until the fever had subsided the dosage was 5 mg/kg orally or intramuscularly. The effective serum phenobarbital level of 15 micrograms/ml was reached with an oral dose of 10 and 15 mg/kg after 25 hours or 90 minutes, respectively, with an intramuscular injection at the same dose after 12 hours and 40 minutes, respectively. These results indicate that the initial dose of phenobarbital in the prevention of further febrile convulsions in children should be 15 mg/kg intramuscularly. Treatment can then be continued orally with 5 mg/kg. PMID- 7472175 TI - [16,16-Dimethyl-prostaglandin E2: protective action on the gastric mucosa]. PMID- 7472176 TI - [Sonographic pancreas diagnosis. Possibilities and limits]. PMID- 7472177 TI - [Pancreatic sonography in undefined upper abdominal symptoms (author's transl)]. PMID- 7472178 TI - [Sonographic demonstration of the pancreatic duct and its clinical relevance (author's transl)]. AB - Out of 19 patients with sonographically demonstrable pancreatic ducts, the Wirsungian duct was found to be dilated on endoscopic retrograde cholangio pancreatography (ERCP) in 18 cases and of normal calibre in only 1 case. The cause of dilatation of the pancreatic duct was chronic pancreatitis in 10 patients and pancreatic carcinoma in 8 patients. There were two differing forms of duct representation in the sonographic picture. Retrospective evaluation of sonographic and ERCP results in 43 patients showed that demonstration of a pancreatic duct of normal calibre is seldom possible using ultrasound. As dilated pancreatic ducts are demonstrable sonographically more readily such a finding must be taken as indicative of pancreatic disease. PMID- 7472179 TI - [Changes of medical in-patients from 1960 to 1977 (author's transl)]. AB - In comparison to 1960 double the number of patients were treated as in-patients in 1977 whereas the duration of stay was reduced by a third. Despite the more advanced age of patients the number of deaths diminished. Many of the present-day in-patients are older than previously. Cardiac arrhythmias, cardiac infarction, arterial vascular disease, diabetes, cancer and alcoholism have increased 2- to 6 fold. PMID- 7472180 TI - [Diagnosis of thymoma and CAT assisted transthoracic biopsy (author's transl)]. AB - Thymomas have an uncharacteristic symptomatology. Their spectrum is greatly enlarged by facultative parathymic syndromes. Despite repeated radiographic investigations thymomas can only be diagnosed with certainty by histological investigation of biopsy material. Method, indications and contraindications of transthoracic biopsy of the anterior mediastinum are demonstrated in 3 cases. This relatively simple method permits diagnosis even in inoperable patients, in whom mediastinoscopy and probatory thoracotomy are contraindicated. PMID- 7472181 TI - [Surgery of stomach ulcer]. PMID- 7472182 TI - [Diabetic amyotrophy--a polygenetic syndrome (author's transl)]. AB - Detailed clinical and biochemical investigations were done in 14 patients with the syndrome of diabetic amyotrophy. Three patients suffered from manifest insulin-dependent diabetes, five had only diminished glucose tolerance. Simultaneously, excretion of creatinine was reduced indicating reduction of healthy muscle mass. In these cases disturbance of glucose tolerance must be considered symptom and not cause of amyotrophy. Further investigations revealed neoplasia (n = 2), generalised amyloidosis (n = 3), polycythaemia vera (n = 1), chronic infectious disease (n = 3), and motor neuron degeneration (n = 3). The attribute "diabetic" must not bea considered representing a single cause in this connection. With treatment aimed at more than one factor, satisfactory results of therapy may be expected in the majority of cases. PMID- 7472184 TI - [Intraoperative autotransfusion]. PMID- 7472183 TI - [Allergic alveolitis following inhalation of mould spores from pot plant earth (author's transl)]. AB - Acute exogenous allergic alveolitis with the typical symptoms of unproductive cough, dyspnoea on exertion, fever, loss of weight, headache and limb pains was observed in a 24-year-old bank employee. Leucocytosis was accompanied by raised ESR; pronounced hypoxaemia and marked restrictive ventilatory defects were found. X-ray changes consisted of severely increased fine reticular interstitial shadowing. The differential diagnosis of allergic alveolitis was confirmed by the demonstration of precipitating antibodies in serum against Pullularia pullulans, Trichoderma viride, Cephalosporium acremonium and Aspergillus fumigatus, moulds with an ubiquitous occurrence. Typical changes of alveolitis were found histopathologically and immunohistologically in the lung tissue after a mini thoracotomy. The source of exposition leading to the disease were pot plant earth and containers in the patient's flat. Moulds could be isolated from these substrates. PMID- 7472185 TI - [Research and planning in social insurance]. PMID- 7472187 TI - [Family therapy--fashion or therapeutic breakthrough]. PMID- 7472186 TI - [Calcium dobesilate for diabetes mellitus?]. PMID- 7472188 TI - [Indications for tonsillectomy--today]. PMID- 7472189 TI - [Clinidine-induced bradycardia: studies of the effect on the human sinus node (author's transl)]. AB - Heart rate, atrioventricular conduction, sinus-node recovery time and effectiveness of carotid-sinus massage on heart rate were measured before and every five minutes after intravenous injection of 0.15 mg clonidine (Catapresan) in 11 patients (averaged age 60 years) with symptomatic bradycardia (dizzy spells, syncope, palpitations). Th results indicated the following significant (P less than 0.05) mean maximal effects: heart rate fell by 12% (from 59 to 52/min), capacity of atrioventricular conduction by 9% (from 132 to 121/min), while maximal sinus-node recovery time ("over-drive suppression") doubled (from 1704 to 3562 ms), heart rates of 120, 150 and 200/min being used for diagnostic atrial overdrive stimulation at each five-minute period after clonidine. During clonidine administration three patients developed a so-called hypersensitive carotid sinus reflex, in two patients the effectiveness of carotid sinus massage decreased, in three if increased, while in three it remained normal. It is concluded that (1) during bradycardia, initiated or increased by sympatholytic drugs, tests should be undertaken to exclude latent sinus-node syndrome and (or) hypertensive carotid-sinus reflex; (2) clonidine should not be used in patients with signs of sinus-node dysfunction. PMID- 7472190 TI - [Danger to health from alcoholic beverages]. PMID- 7472191 TI - [Flexible endoscopy with a piggyback dilatator]. PMID- 7472192 TI - [Cranial operations in Kenya]. PMID- 7472193 TI - [Surgery of Crohn's disease: a study of 155 patients after intestinal resection (author's transl)]. AB - Operative mortality after intestinal resection in 155 patients with Crohn's disease was 8.4%. In 12 of 13 the cause of death was septic complications, most of them related to prolonged pre-operative cortisone medication. Follow-up examination was possible in 90% of all discharges. Five-year recurrence rate was 37.6%. Recurrence was not prevented by long-term drug treatment. Purely macroscopic assessment of the resection margins would have led to excision within diseased gut: for this reason frozen sections are recommended before the anastomosis is made. Primary death-rate in 30 cases of colectomy with primary ileorectostomy was 3%. In three patients the rectum had to be excised, while in one the anastomosis had to be taken down because of recurrence. The failure rate was thus only 11%. If rectoscopy, stepwise biopsy and intra-operative frozen section indicate a normal rectum it is recommended that ileorectostomy be done rather than diversion ileostomy with occlusion of the rectum or even proctocolectomy. PMID- 7472194 TI - [Effect of clonidine on plasma-catecholamine levels at rest and on ergometric exercise (author's transl)]. AB - The effect of clonidine on peripheral venous-plasma catecholamine concentration was assessed in ten subjects with normal circulation, at rest and on submaximal bicycle exercise. Three hours after oral intake of 300 micrograms clonidine in single dose there was significant lowering of the plasma adrenaline and noradrenaline levels. Six hours after intake the lowest plasma catecholamine level was reached, but after 24 hours the clonidine effect had largely disappeared. The sympatholytic effect of clonidine was relatively less marked with bicycle exercise than at rest. When clonidine had been given for only a short period at moderate dosage, there was no exaggerated catecholamine rise. Clonidine administration did not effect plasma dopamine levels. These results indicate that clonidine decreases plasma adrenaline and noradrenaline levels without significant effect on circulatory regulation. PMID- 7472195 TI - [Direct M-mode and two-dimensional echocardiographic demonstration of acute pulmonary arterial embolism (author's transl)]. AB - Two weeks after hysterectomy, circulatory collapse with hypotension and tachycardia occurred in a 40-year-old woman. M-mode and 2-D echocardiography confirmed the clinical diagnosis of acute pulmonary arterial embolism. PMID- 7472196 TI - [Therapeutic problems in chronic active hepatitis]. PMID- 7472197 TI - [Acute kidney failure after major surgery]. PMID- 7472199 TI - [Acupuncture]. PMID- 7472198 TI - [Percutaneous liver biopsy under sonographic control]. PMID- 7472200 TI - [Heterologous insemination]. PMID- 7472201 TI - [Enteritis caused by atypical Yersinia (author's transl)]. AB - For the first time in the Federal Republic of Germany Yersinia species "frederiksenii", "kristensenii", "serovar 0 : 6" (each once) and "serovar 0 : 5" (twice) were found in the stool of 5 patients with gastroenteritic complaints. Serum agglutinins against these species could not be demonstrated. Other enteropathogenic microorganisms were excluded microscopically, by culture and serologically. These Yersinia isolates were resistant against penicillin G, ampicillin, carbenicillin, ticarcillin, and cefalotin. PMID- 7472202 TI - [Hypernatraemic hyperosmolar syndrome (author's transl)]. AB - Between 1972 and 1979 34 patients with severe hypernatraemia (serum sodium larger than or equal to 160 mmol/l) were admitted to the intensive care unit. Among these there were 45% of neuropsychiatric patients with reduced consciousness. Maximal serum sodium concentration was 168 (160--204) mmol/l, maximum serum osmolality 391 (340--520) mosm/kg H2O. Treatment consisted of infusion of hypotonic solutions and potassium substitution. Fluid balance was positive (5,9 during 43 hours) until normal serum sodium concentrations had been reached. The mortality of 77% was exclusively due to the underlying diseases. PMID- 7472204 TI - [Manifestations of porphyria cutanea tarda]. PMID- 7472203 TI - [Freedom of teaching, domestic authority and medical confidentiality of the university instructor]. PMID- 7472205 TI - [After blunt abdominal trauma: peritoneal lavage or sonography?]. PMID- 7472206 TI - [Diagnostic peritoneal lavage]. PMID- 7472207 TI - [Psychosomatic aspects of intensive medicine]. PMID- 7472208 TI - [Accuracy of endoscopic retrograde pancreatico-cholangiography in the diagnosis of pancreatic disease: an analysis of three thousand investigations (author's transl)]. AB - Endoscopic retrograde pancreatico-cholangiography (ERPC) gave the correct diagnosis in 83% of 269 cases of pancreatitis confirmed by operation, clinical features, laboratory findings and course. It provided the correct diagnosis in 85% of 82 cases of necrosis of the pancreas and pseudocysts confirmed at operation. ERPC is one of the most important methods of investigation in papillary stenosis, because it can demonstrate both ducts and can reveal functional changes, especially delayed emptying. The correct diagnosis of carcinoma of the pancreas, confirmed at operation or histologically, was made in 88% of 107 cases. Endoscopy correctly diagnosed 26 of 27 cases of papillary carcinoma. Biopsy with subsequent histological study and cytological analysis of pancreatic secretion further served to confirm the diagnoses established by ERPC. PMID- 7472209 TI - [Staging laparotomy for malignant lymphoma: interdisciplinary aspects and results (author's transl)]. AB - Between 1963 and 1979 a total of 357 patients with malignant systemic lymphatic disease underwent splenectomy in the course of exploratory laparotomy. There were three groups: 1. Hodgkin's disease (215 patients), 2. non-Hodgkin's lymphoma (93), and 3. chronic lymphoid leukaemia (49). Comparing pre- and postoperative findings, primary exploratory laparotomy remained the most exact foundation for the differentiated treatment of Hodgkin's disease in stages I-III. Systematic staging laparotomy was of less importance for non-Hodgkin's lymphoma and chronic lymphoid leukaemia because of the much more frequent invasion of bone marrow, spleen and liver. However, it should be considered more often in the management of such cases. PMID- 7472211 TI - [Chronic hepatic porphyria with uroporphyrinogen decarboxylase defect in four generations (author's transl)]. AB - A 54-year-old patient was found to have clinically manifest chronic hepatic porphyria (porphyria cutanea tarda) with characteristic skin findings and pathological porphyrinuria with dominance of uro- and heptacarboxyporphyrin in combination with only slight liver damage. Four generations of his family were investigated and an autosomal dominantly inherited uroporphyrinogen decarboxylase defect was found. In children and grandchildren the urinary porphyrin biochemistry also showed a chronic porphyrin metabolic disorder, in some cases with transition to the initial stages of chronic hepatic porphyria. The patient was treated with metabolic alkalisation using Uralyt-U. Skin changes became normal within 6 months. The chronic hepatic porphyria regressed through varyingly severe latent stages over two years until normal porphyrin in urine was reached. PMID- 7472210 TI - [Pancreatic lipase: reference values at 25 degrees C (author's transl)]. AB - Development of a new specific test for estimation of pancreatic lipase activity (EC 3.1.1.3.) required establishment of reference values. An upper reference range of 170 to 190 U/l at 25 degrees C was determined from a group of 368 females and 435 males aged 20 to 70 years. Values below 170 U/l are very likely to be normal, values above 190 U/l are considered pathological. Values between 170 and 190 U/l require further controls. For practical reasons age and sex dependency need not be accounted for as these influences are negligible. PMID- 7472212 TI - [Ideas about cancer]. PMID- 7472214 TI - [Determination of sinuatrial conduction time in hospital]. PMID- 7472213 TI - [Digestive function of the residual pancreas after partial duodenopancreatectomy for chronic pancreatitis (author's transl)]. AB - Forty-eight patients with chronic recurrent pancreatitis treated by resection of the head of the pancreas were restudied more than a year post-operatively. In addition to general features such as symptoms, alcohol consumption and work ability, faecal weight, its fat content, fat and fatty acid balance and faecal chymotrypsin were measured. According to the patients' own estimate, late results were good or very good in 70-90%. Faecal fat content and balance indicated high grade exocrine pancreatic insufficiency in 80 and 90%, respectively. But it was easily controlled by drugs. In a third of the cases there was the need to supplement the diet with medium-chain triglycerides. Its components are satisfactorily absorbed even when the fat utilisation is severely abnormal. PMID- 7472215 TI - [Prohibition of competition between physicians]. PMID- 7472216 TI - Digoxin dose precision: prescribing aids or intuition? PMID- 7472217 TI - Diarrhoea: causes and specific treatment. PMID- 7472218 TI - Medication and pregnancy. PMID- 7472219 TI - [Lymph node toxoplasmosis]. PMID- 7472220 TI - [Respiratory damage in patients with burns]. PMID- 7472221 TI - [Pneumatosis cystoides intestinalis]. PMID- 7472222 TI - [Eosinophilia and helminthiasis]. PMID- 7472223 TI - [Diabetic diet]. PMID- 7472225 TI - [Experimental animals in research]. PMID- 7472224 TI - [Artificial sweeteners in the diabetic diet]. PMID- 7472227 TI - [Risk of infection associated with umbilical vein catheterization and exchange transfusion in neonatal hyperbilirubinemia]. PMID- 7472226 TI - [Rapid and specific determination of serum HCG in gynecological emergencies]. PMID- 7472228 TI - [Problems in the choice of tranquilizing agents]. PMID- 7472229 TI - [Paraparesis as an initial sign in systemic lupus erythematosus]. PMID- 7472230 TI - [Treatment of colpitis]. PMID- 7472231 TI - [Clinical pathological conference LX. Progressive neurological symptoms and changes shown by computerized tomography]. PMID- 7472232 TI - Fatty acid accretion in the development of human spinal cord. AB - Lumbar and cervical segments of fetal and infant spinal cord were examined to determine the deposition rate of individual fatty acids in the spinal cord during the last trimester of fetal development and first 16 weeks of life. Fatty acid accretion rates were determined by regression analysis of spinal cord fatty acids at varying developmental ages. During the last trimester of fetal growth, accretion of major tissue fatty acids occurred earlier in cervical segments of spinal cord and at a greater rate than observed in lumbar regions. Postnatal accretion of fatty acids in cervical regions of spinal cord was minimal on a per gram of tissue basis with the exception of moderate increases in total omega-9 and saturated fatty acids, reflecting myelination. Significant postnatal accretion of fatty acids in lumbar regions of the spinal cord occurred some 4 weeks postnatally at rates significantly in excess of in utero rates. These developmental changes in fatty acid accretion are quantitatively relevant to estimates of fatty acid incorporation into spinal cord and parallel known processes of myelination and increasing neuro-motor activity in the developing infant. PMID- 7472233 TI - Assessment of individual differences among healthy newborns on the Brazelton scale. AB - Inter-individual differences and behavioral stability assessed with the Brazelton scale were examined in twenty 3-8-day-old full-term newborns at a German newborn nursery. The extent of variability measurable with Brazelton scores was determined by the relative mean information value "Hrel" in each variable, which indicates the discriminating power of each scale. The value "Hrel" was high in irritability, consolability, cuddliness and alertness. Inter-individual differences, illustrated by individual mean scores, were lowest in orientating behaviors. Differences in alertness correlated with several other reaction patterns. No significant correlations were found between the NBAS variables and neonatal sex, birth weight, birth duration, and maternal age. However, the relative weight gain between day 1 and day 8 was correlated with alertness, orientation to face and to rattle, irritability, rapidity of build-up and state lability. Individual behavioral stability over time was generally low in most variables. The low degree of behavioral stability and the low relative mean information in several variables reduce the applicability of the scale for the originally suggested purposes. PMID- 7472234 TI - Clinical assessment of gestational age in the newborn infant. Comparison of two methods. AB - A clinical assessment of gestational age was performed in a group of 92 newborn infants. The Dubowitz method (1970) based on 10 neurological and 11 "external" criteria and the Finnstrom method (1977) based on 7 "external" criteria have been applied. A comparison between these two methods showed a correct estimation of gestational age within +/- 2 weeks in 86.95% of cases with the Finnstrom method and in 80.72% of cases with the Dubowitz method. A correct estimation within +/- 3 weeks was found in more than 90% of the infants with both methods. No significant difference between the two methods was found. In about 10% of cases of Dubowitz method could not be applied because of the serious conditions of the infants. Assessment of gestational age with the Finnstrom method proves to be less stressful for the infant, less time consuming and feasible on a larger number of infants than the assessment with the Dubowitz method. For these reasons the Finnstrom method seems to be more advisable for clinical routine work. PMID- 7472236 TI - How long should a breast feed last? AB - Patterns of milk flow were studied in 50 mothers who were breast-feeding normal birth-weight babies on days 5-7 of the puerperium by progressive test weighing at 5-min intervals during two consecutive feeds. Compared with a regime in which mothers attempted to feed for 10 X 10 min on alternate breasts, a regime of 5 X 5 X 5 X 5 min increased the amount of milk taken in the first 10 min did not influence the final milk intake of the suckling-induced prolactin release. The wide variation of breast-feeding patterns between mothers was demonstrated in respect of the duration of the feed (mean 17.3 min; S.D. +/- 3.1; range 7-30 min), the initial rate of milk flow (mean 6 g/min; S.D. +/- 4.2, range 1-14 g/min) and the final milk intake (mean 70.9 g; S.D. +/- 20.5, range 42-125 g). The advice to breast-feed for 20 min was in appropriate for the majority of mothers because the nutritive feeding time was 15 min or less in 75% of the feeding episodes. The milk intake correlated with the initial rate of milk flow but not with the duration of the feed, the infant's birth weight, or the time since the last feed. It is suggested that the duration of a breast feed should be determined by the infant's response and not by an arbitrary time schedule. PMID- 7472235 TI - Fatty acid accretion in fetal and neonatal liver: implications for fatty acid requirements. AB - The fatty acid content of liver was determined during the last trimester of infant growth and first 4 months of life in order to estimate fatty acid levels of developing liver and to assess minimal fatty acid requirements for tissue synthesis. Accretion rates were computed by regression analysis on fatty acid determinants of total liver lipid extracts from infants of varying developmental ages. During the last trimester of liver growth, accretion of most fatty acids paralleled increases in liver weight with the exception of C18:3, omega-3. This fatty acid remained at consistently low levels during the last trimester and represented less than 10% of total omega-3 fatty acids present in liver, even though significant accretion of longer chain omega-3 homologues occurred during the final trimester. For the term infants studied, significant increase in liver weight did not occur during the early weeks of life. However, liver levels of major fatty acids declined during the first five weeks of life apparently reflecting mobilization of saturated, omega-9 and omega-3 fatty acids from liver. During the first four months of life C18:2, omega-6 and total omega-6 fatty acid content of liver increased about 3.5- and 2-fold, respectively. After the initial 5 week lag net accretion of other fatty acid components also occurred in the developing liver. These developmental changes in fatty acid components of liver are quantitatively relevant to estimating the magnitude of the potential essential fatty acid reserve that may be present in liver of the developing human neonate. PMID- 7472237 TI - Effects of father-infant postpartum contact on their interaction 3 months after birth. AB - One group of fathers were allowed to handle their newborn infant immediately after its Caesarean-section delivery, while another group were not permitted to do so. In a play situation performed 3 months later, the contact-group showed more touching behaviour towards their infants than the other group. PMID- 7472238 TI - Human fetal eye movements: detection in utero by ultrasonography. AB - Eye movements of the human fetus were detected in utero by means of real-time B scan and M-mode ultrasonography. In 14 low-risk fetuses studied between 32 weeks menstrual age and term either rapid or slow eye movements or a mixture of both types were detected in 60 +/- 10% (mean +/- SD) of 558 2 min scoring epochs. A comparison of direct echoscopic detection of eye movements in a 3-day-old neonate confirmed that the movements observed in the orbital region with the real-time scanner corresponded well with eye movements seen directly. Thus eye movements can be added to the list of phenomena which can be observed noninvasively for the study of human fetal behaviour. PMID- 7472239 TI - Industrial chemical safety testing: one company's approach. PMID- 7472240 TI - A multigeneration fish toxicity test as an aid in the hazard evaluation of aquatic pollutants. PMID- 7472242 TI - Methods of dose evaluation for incorporation of radionuclides: applicability to environmental chemicals. PMID- 7472241 TI - Risk/benefit analysis for toxic chemicals. PMID- 7472243 TI - The International Register of Potentially Toxic Chemicals (IRPTC). PMID- 7472244 TI - A proposed approach to chemical fate assessments. PMID- 7472245 TI - Structure-activity relationships and additivity in fish toxicities of environmental pollutants. PMID- 7472246 TI - The experimental approach to teratogenicity. PMID- 7472247 TI - Basis concepts of radiation protection. PMID- 7472248 TI - Experiences with the test scheme under the chemical control law of Japan: an approach to structure-activity correlations. PMID- 7472249 TI - The yardstick concept for the hazard evaluation of substances. PMID- 7472250 TI - Acceptability of data from other countries. PMID- 7472251 TI - Effect of combinations of Irideae carrageenan and cellulose on the absorption of some nutrients from the alimentary tract of rats. PMID- 7472252 TI - Cancer incidence among 5686 asbestos-cement workers followed from 1943 through 1976. PMID- 7472253 TI - Lead in Greenland snow. PMID- 7472254 TI - The teratogenic and mutagenic effects of dinitro-o-cresol-containing herbicide on the laboratory mouse. PMID- 7472256 TI - Recent activities of the "GESAMP" working group on the evaluation of hazardous substances carried by ships. PMID- 7472255 TI - A correlation study of biodegradability determinations with various chemicals in various tests. II. Additional results and conclusions. PMID- 7472257 TI - Accumulation and depuration of metals by duckweed (Lemna perpusilla). PMID- 7472258 TI - Aryl hydrocarbon hydroxylase in pigeon skin and its possible relevance to monitoring air pollution. PMID- 7472259 TI - Tetrahydro-beta-carbolines elevate plasma prolactin in male rats. AB - The effects of systemic administration of three tetrahydro-beta-carbolines (TH beta C's) on plasma prolactin were determined in male rats. After intraperitoneal administration, 6-methoxy-TH beta C (6-MeO-TH beta C; 80 mg/kg) significantly elevated prolactin levels for 6 min. After intravenous administration via an indwelling jugular cannula, 6-MeO-TH beta C (20 mg/kg but not 2.0 mg/kg), TH beta C (20 mg/kg) and 6-OH-TH beta C (20 mg/kg) all produced a significant elevation in plasma prolactin for 30 min. with the latter compound effective for 60 min. Previous evidence indicated that the TH beta C's interact relatively selectively with the serotonergic neurotransmitter system to increase serotonergic function. Thus, the present data are consistent with previous studies which indicated that increased serotonergic activity resulted in enhanced prolactin secretion. PMID- 7472260 TI - Effect of testosterone on the androgen receptor in the liver cytosol and nuclei of castrated male rats. AB - The androgen-binding sites in the cytosol and nuclei were decreased 1 week following castration and this reduced level was sustained for 2 weeks. A maximal increase of the androgen-binding sites in the cytosol was induced by a single injection of testosterone, while the sites in the nuclei were increased gradually with the duration of testosterone injection. These findings suggest that testosterone was able to induce the binding sites in the liver cytosol and nuclei which had been reduced after castration. PMID- 7472261 TI - On the mode of the prolactin release-inhibiting action of the serotonin receptor blockers metergoline, methysergide, and cyproheptadine. AB - Using animals with large electrolytic lesions of the median eminence-mediobasal hypothalamus, we confirmed earlier findings that metergoline (ME) and methysergide (MS) inhibit PRL secretion through activation of the dopamine receptors of the pituitary lactotrophs and established, in a quantitative manner, that their dopaminergic potencies are comparable to the potency of the dopamine receptor agonist, piribedil, with ED50 in the order of 0.35 to 0.22 mg/kg. Cyproheptadine (CYP), acting by an unknown mechanism, had only a weak inhibiting effect (ED50 greater than 20.0 mg/kg) in these experimental conditions. In the second part of the study, the PRL-inhibiting actions of ME, MS, CYP, and piribedil, respectively, were tested against the PRL release-stimulating effect of activation of the central serotonergic system that was induced by administration of a large dose of L-5-hydroxytryptophan (5HTP; 100 mg/kg), a small dose of 5HTP (15 mg/kg) in rats pretreated with fluoxetine, or by the serotonin receptor agonist quipazine (10.0 mg/kg, ip). The inhibiting potencies of ME (ED50 0.019, 0.014, and 0.048 mg/kg, respectively) against these three stimuli were much larger than in the lesioned animals or than the corresponding potencies of piribedil (ED50 2.2, 0.24, and 0.41 mg/kg, respectively). It is assumed that in these experimental conditions ME inhibited PRL release by blockade of the central serotonin receptors in addition to its dopaminergic effect and that at low doses (0.1 mg/kg or less) the entire inhibiting effect of ME was probably due to its antiserotonergic activity. With MS, which is a weaker serotonin receptor blocker than ME (ED50 0.178, 0.075, and 0.55 mg/kg, respectively, for the three serotonergic stimuli of PRL release), the antiserotonergic component in its PRL-inhibiting effect was evident but less clearly separable from the dopaminergic component in experiments with 5HTP and with fluoxetine plus 5HTP, whereas in experiments with quipazine the entire action could be accounted for by its dopaminergic activity. CYP was the least potent among the three blockers (ED50 0.6, 0.4, and 1.37 mg/kg, respectively, for the three serotonergic stimuli of PRL release), but appropriate tests indicated that it acted only as a serotonin receptor blocker and not by virtue of its antihistaminic, anticholinergic properties or by a direct action on the pituitary lactotrophs. SQ 10,631, another serotonin receptor blocker that was also tested, had no PRL-inhibiting activity. Because of the dual nature of the PRL-inhibiting mechanism of ME and MS and the low effectiveness of CYP, combined possibly with other actions, the serotonin receptor blockers have limited value in studies concerning the role of the central serotonergic system in the regulation of PRL secretion. PMID- 7472262 TI - Stimulation of gluconeogenesis by cortisol in fetal rat liver in organ culture. AB - Fetal rat liver explants (gestational day 20) depleted of glycogen converted L [14C]alanine into labeled glucose and glycogen. Cortisol 1) stimulated net glyconeogenesis about 2-fold at 22 h of incubation after a 7-h lag; 2) increased the total amount of glycogen in the explant 2-fold; 3) stimulated the net incorporation of label from [14C]alanine into both glucose and glycogen. Both [3H]glucose added to the medium and glucose generated by gluconeogenesis from [14C]alanine contributed to the glycogen formed. Cortisol led to greater 14C uptake than 3H uptake into glycogen. During gluconeogenesis, [14C]glycogen was first formed and subsequently broke down to [14C]glucose. In the absence of cortisol, glucose and glycogen reached equilibrium with each other. In the presence of cortisol, preferential uptake of alanine into glycogen persisted throughout the 22 h of incubation, and equilibrium was not reached. Gluconeogenesis from [14C]glycerol was greater than from [14C]alanine. Cortisol stimulated its conversion to glycogen but not to glucose. It was concluded that gluconeogenesis in fetal rat liver occurs under organ culture conditions, that its stimulation by cortisol is direct and is localized in two regions: conversion of L-alanine to triose phosphates and of glucose-6-phosphate to glycogen. PMID- 7472263 TI - Retarded growth rate caused by glucocorticoid treatment or dietary restriction: associated changes duodenal, jejunal, and ileal calcium absorption in the chick. AB - Calcium (Ca) absorption is reportedly stimulated in the ileum of glucocorticoid treated rats. Our study was designed to investigate this phenomenon in the chick; to determine to what extent any stimulation might compensate for inhibition of Ca absorption in the duodenum; and to compare any changes seen with those caused by growth retardation induced by dietary restriction alone. We assessed CA absorption from different sites in the intestine of chicks treated with betamethasone (BM; 25 micrograms/kg . day for 14 days), or diet-restricted to permit growth at the same rate as those treated with BM, using the ligated loop technique in vivo. There was a similar significant inhibition of Ca absorption in both BM-treated and diet-restricted chicks in both duodenum (to 73% and 67% of the control level, respectively) and jejunum (to 74% and 77% of the control level, respectively). Absorption of Ca from the ileum of BM-treated birds equaled that in controls, but diet-restricted chicks unexpectedly showed a 100% stimulation. We conclude that in the chick, BM treatment does not stimulate ileal calcium absorption, but diet-restriction does. The lack of effect of bM in the ileum may reduce the magnitude of the decrease in Ca absorption from the entire small intestine when compared to the duodenum and jejunum alone, but the stimulated absorption seen in the ileum of diet-restricted birds is almost sufficient to restore Ca absorption from the entire small intestine to the control level. PMID- 7472264 TI - Tumor-promoting phorbol esters affect production of prolactin and growth hormone by rat pituitary cells. AB - GH4C1 cells are strain of rat pituitary cells which synthesize and secrete both PRL and GH. Phorbol esters, which are potent tumor promoters in mouse skin and have diverse actions in a variety of cell and tissue systems, were found to alter hormone production by GH4C1 cells. 12-O-Tetradecanoylphorbol-13-acetate (TPA), the most potent phorbol ester, stimulated both PRL synthesis and release. The synthesis of PRL was stimulated 2.3- to 6.2-fold over control values after 3 days of treatment; the ED50 for TPA was 8.9 +/- 2.1 nM (5.5 +/- 1.3 ng/ml). The release of stored PRL was stimulated within 10 min of the addition of 100 ng/ml TPA to the culture medium. Maximum stimulation of release was 1.3- to 2.3-fold over control, and the ED50 for TPA was 13.2 +/- 2.3 nM (8.1 +/- 1.4 ng/ml). Another tumor-promoting phorbol ester, phorbol 12,13-didecanoate (PDD), stimulated PRL synthesis and release to the same maximum extent at TPA, but two biologically inactive compounds, 4 alpha-PDD and phorbol, had no effect. The stimulation of PRL release by TPA was a Ca++-dependent process; Co++, a competitive antagonist of Ca++, at 2.0 mM blocked completely the stimulation of PRL release by 100 ng/ml TPA, and this block was overcome by 2.0 mM Ca++, but not by 2.0 mM Mg++. Although phorbol esters stimulate prostaglandin (PG) synthesis in certain other cells types, PGs were not required for TPA action in GH4C1 cells. Indomethacin, a PG cyclooxygenase inhibitor, at 200 ng/ml did not inhibit the stimulation of PRL synthesis and release by TPA. Furthermore, there was no detectable (less than 50 pg/ml) PGE2 or PGF2 alpha in the medium of control or TPA-treated cells. In GH3 cells, a related cell strain, TPA (100 ng/ml) decreased GH production in both control and hydrocortisone-stimulated cells to 35% of untreated control values after 3 days of incubation. TPA also altered cell morphology, but had little or no effect on cell growth. We conclude that GH cells provide a useful system to study the mechanisms of phorbol ester action on differentiated functions. PMID- 7472265 TI - Multihormonal regulation of the estrogen receptor in rat liver. AB - Hepatic estrogen receptors were measured in ovariectomized female rats using isoelectric focusing in polyacrylamide gel. Hypophysectomy or adrenalectomy reduced the receptor level to approximately 10% and 42%, respectively, of the control value. Treatment of hypophysectomized (Hx) rats with dexamethasone alone was without effect. When Hx rats were given a pituitary transplant under the kidney capsule in order to induce liver estrogen receptors, dexamethasone treatment more than doubled the inductive effect of the pituitary transplant. The administration of human GH (hGH) in osmotic minipumps to Hx rats led to an induction of hepatic estrogen receptor levels to 48% of the control value. When hGH infusion was combined with dexamethasone treatment, a complete restoration of the receptor level to that in control rats was seen. When administered in minipumps, ovine PRL (oPRL) and bovine GH (bGH) as well as a combination of oPRL and bGH led to an induction of estrogen receptors in Hx rat livers when dexamethasone treatment was given simultaneously. However, oPRL, bGH, and oPRL plus bGH failed to give a complete restoration of hepatic estrogen receptor levels. It is concluded that the estrogen receptor in rat liver is under multihormonal control and that glucocorticoids in combination with a pituitary factor(s) with similar properties to hGH seem to be responsible for the maintenance of the normal level of the receptor in female rats. Possibly, both GH and PRL are involved in the control of the receptor. PMID- 7472266 TI - Elevation of serum 3,5,3'-triiodothyronine and thyroxine levels in rats fed Remington diets; opposing effects of nutritional deficiency and iodine deficiency. AB - In the course of experiments on iodine deficiency induced by Remington diets in rats, we observed that the Remington diet supplied by ICN Nutritional Laboratories, though very deficient in iodine (less than 20 micrograms I/kg), did not lead to the rapid loss of thyroid iodine and the rapid decrease in serum T4 expected on the basis of previous studies with a similarly iodine-deficient Remington diet from another source. In searching for an explanation for this observation, we noted that the ICN Remington diet was nutritionally much more deficient than Remington diets from other suppliers. We also noted that when rats were placed on the iodide-supplemented ICN Remington diet there was a marked increase in serum T3 and T4. In one experiment, rats receiving the ICN Remington diet plus KI in the drinking water for 16 days showed a serum T3 level of 109 +/- 16 ng/dl and a serum T4 level of 6.6 micrograms/dl compared to 52 +/- 7.8 and 4.4 +/- 0.8, respectively, in control rats on a stock diet. These elevations were not simply the result of increased binding to serum proteins. Serum protein-binding studies by the method of equilibrium dialysis showed a very slight decrease in the percent dialyzable fraction for T3 and T4. However, calculated free T3 levels were significantly elevated (P less than 0.001), and increases in free T4, though less striking, were also significant. These elevations were not accompanied by evidence of hyperthyroidism, as judged by measurements of O2 consumption or serum TSH. Although the specific nutritional factors and mechanisms have not yet been defined, our studies demonstrate that nutritional deficiencies in a Remington diet may act to oppose the effects of the iodine deficiency itself. Our observation that the iodide-supplemented ICN Remington diet has a marked serum T3 and T4-elevating effect offers a possible explanation for the blunted thyroidal responses of rats to the same diet lacking added iodide. Our studies also suggest that alteration of peripheral T4 and T3 conversion may not be the only mechanism by which nutritional factors affect serum T3 and T4 levels. PMID- 7472267 TI - Phenolic and nonphenolic ring iodothyronine deiodinases from rat thyroid gland. PMID- 7472268 TI - Autofeedback regulation of prolactin (PRL) secretion: effect of prolactin before suckling on the subsequent nursing-induced release of PRL in the lactating rat. AB - PRL can have an inhibitory effect upon its own release. The present investigation assessed quantitative and temporal relationships among exogenously raised systemic PRL concentrations, PRL discharged in situ, and PRL negative feedback control of PRL release. Lactating rats were administered PRL in amounts designed to increase systemic plasma PRL concentrations to levels that either exceed (3 mg ovine PRL injected sc) or fall within (five iv injections of 400 or 800 ng rat PRL given at 1-min intervals) the range of PRL concentrations normally produced by suckling stimulation. The 800-ng iv or 3-mg sc dose of PRL did not block the suckling-induced rise of plasma PRL concentration when given 30 min before suckling. When administered 4 h beforehand, however, 3 mg PRL reduced both the suckling- and ether-stimulated rises of plasma PRL. Although the 400- or 800-ng iv dose of PRL increased systemic plasma PRL concentrations to levels (280 and 389 ng/ml, respectively) that approximate those of the suckled rat, neither dose inhibited the suckling-induced release of PRL when given 4 h before nursing. These data indicate that high concentrations of PRL are required before PRL from the systemic circulation can activate PRL autoregulatory mechanisms, and that these mechanisms are not activated as an immediate response to rising plasma PRL titers. The suckling-induced release of PRL was also inhibited by 3 mg PRL given 16 h before nursing. The PRL discharged in situ by 15 min of nursing was not effective, however, in blocking the suckling-induced release of PRL 16 h later. Since large amounts of PRL can presumably reach PRL regulatory mechanisms during in situ release via retrograde pituitary secretion, we conclude that the functional significance of autofeedback control of PRL release may be limited to conditions where PRL secretion is prolonged or at an abnormally high rate. PMID- 7472269 TI - Enzymatic conversion of the major polypeptide chains of thyroglobulin. AB - The subunit composition of fully reduced bovine thyroglobulin consists of two polypeptide chains with molecular weights near 300,000. These two chains have been separated on sodium dodecyl sulfate gels by electrophoresis and were referred to as S and F. The effect of incubating native bovine thyroglobulin with a commercial preparation of horseradish peroxidase resulted in the conversion of S to F (and faster migrating polypeptides). Since this reaction was independent of the presence of H2O2 and iodide, it was necessary to demonstrate that the activity was present in the peroxidase enzyme which could be identified by its heme group. When the enzyme preparation was fractionated on Sephadex G-100, the activity responsible for the conversion of S to F was not associated with the heme peak but with another protein peak which eluted later on the column. Proteolytic inhibitors, such as phenylmethyl sulfonyl fluoride and pancreatic trypsin inhibitor, caused partial inhibition of the activity responsible for the conversion of S to F, whereas inhibitors of peroxidase activity had no effect. Guinea pig thyroglobulin was less susceptible to the proteolytic activity present in the peroxidase preparation than bovine thyroglobulin. The subunit composition of guinea pig thyroglobulin is different from that of bovine, since three bands of different sizes are present. The 300,000 mol wt subunit of guinea pig thyroglobulin (band A) ws largely degraded, whereas the 210,000 mol wt subunit (band B) was partially degraded and the 100,000 mol wt subunit (band C) was unaffected. PMID- 7472270 TI - An ovarian role in prolonging and terminating the two surges of prolactin in pseudopregnant rats. AB - Daily nocturnal (N) and diurnal (D) surges of PRL are characteristic of a pseudopregnancy (psp) induced by cervical stimulation (CS). These semicircadian surges persist for 13-14 days, after which time they cease, and psp ends. The purpose of this study was to describe the involvement of the ovarian steroids in the maintenance and termination of the N and D surges of PRL. CS of long term ovariectomized (OVX) rats resulted in N and D surges of PRL lasting 14 days. By day 16 after CS, neither surge of PRL was present. The sc placement of Silastic implants containing large quantities of progesterone in long term OVX-CS rats sustained and enhanced the N surge of PRL until at least day 16. The D surge was also sustained until day 16, but its magnitude was normal. Estradiol alone was capable of maintaining the D but not the N surge for this same time interval. However, pharmacological quantities of estradiol restricted the magnitude of the N surge generated in the progesterone-implanted animal. Since neither surge of PRL was present by day 14 after CS in intact rats, but at least the nocturnal surge was still present at this time in long term or acutely OVX rats, this implies an active role of ovarian steroids in the termination of PRL surges at the end of psp. At the end of psp, luteal progesterone secretion declines and follicular estradiol secretion increases. Varying sizes of Silastic implants containing progesterone or estrogen were placed into psp animals after acute ovariectomy on day 9 to approximate this ovarian steroid secretory pattern of intact psp rats. Only when the decrease in progesterone was coupled with a modest increase in estrogen was there a total cessation of the two daily surges of PRL in OVX-CS rats. The data suggest that the PRL surges end in the intact psp rat due to the waning response to CS as well as the shifting steroid ratio represented by the fall in progesterone in the presence of a small amount of estrogen. PMID- 7472271 TI - Effects of human chorionic gonadotropin in the rabbit corpus luteum: loss of estrogen receptor and decreased steroidogenic response to estradiol. AB - The effects of hCG on luteal estrogen receptor and steroidogenesis were examined in 9-day-pseudopregnant rabbits. Twenty-four hours after the injection of ovulatory dosages of hCG (10-100 IU), a dose-related loss of luteal estrogen receptor and in vitro progesterone production was observed. The declining progesterone production was not due to the increased conversion of progesterone to the major metabolite, 20 alpha-dihydroprogesterone. The loss of steroidogenesis induced by hCG was not permanent and could be reversed by estradiol treatment started within 24 h of hCG injection. In those animals with a higher luteal estrogen receptor content at the start of the estrogen treatment, the steroidogenic response was greater. These findings indicate that hCG-induced loss of steroidogenesis is associated with the loss of luteal estrogen receptor. The degree of restoration of progesterone synthesis induced by a 24-h period of estradiol treatment may be related to the content of unoccupied cytoplasmic estrogen receptor in the corpus luteum. PMID- 7472272 TI - Ovine maternal and fetal plasma oxytocin concentrations before and during parturition. AB - To study the relationships between fetal and maternal oxytocin (OT) levels and the initiation of labor in sheep, paired maternal and fetal plasma OT concentrations (microunits per ml) were measured by RIA. Samples were obtained daily from pregnant ewes and their fetuses for 5 days before spontaneous delivery and frequently during the first and second stages of labor and during the 3 h after delivery. The mean maternal plasma OT concentration during the first stage of labor was not different from that preceding labor. In contrast, the mean maternal plasma OT level during stage 2 of labor was significantly higher than the earlier baseline maternal values or the mean paired fetal concentration. There was no significant increase in the mean fetal plasma OT concentration before delivery. The newborn plasma OT concentration was elevated 15 min after delivery. From these data, we conclude that in the sheep, 1) the onset of labor is not associated with increased maternal plasma OT levels, 2) cervical or vaginal distension may be the stimulus for maternal OT release during stage 2 of labor, 3) an increase in the fetal plasma OT concentration does not occur before the initiation of labor or during the course of labor, and 4) stress in the final moments of labor or in early neonatal life may be responsible for elevated cord and early neonatal plasma OT levels. PMID- 7472273 TI - Human breast tumor estrogen receptor: effects of molybdate and electrophoretic analyses. AB - The largest and smallest discrete forms of the estrogen receptor in human breast tumor cytosol were characterized by competitive steroid binding, ultracentrifugation, gel filtration, and electrophoresis in polyacrylamide gels of several concentrations. Incubation of cytosol with [3H]estradiol and centrifugation in glycerol gradients containing 20 mM Na2MoO4 and 0 or 150 mM KCl revealed a 9-10S form of the receptor. It resembles the molybdate-stabilized complexes in cytosols of other human and rodent, malignant and healthy tissues, and the complex detected in breast tumor cytosol containing leupeptin, a bacterial protease inhibitor. Preservation of receptor integrity during purification and discrimination from serum steroid-binding components are facilitated by inclusion of molybdate in all buffers. Possible mechanisms of action of molybdate include the inhibition of ribonuclease action on RNA associated receptor forms and protection against specific proteolytic cleavage by stabilization of a phosphate group on the vulnerable residue or a neighboring one. During fractionation of tumor cytosol in the absence of molybdate, the receptor is converted to a mixture of fragments. The smallest that retains the bound steroid, the mero-receptor, resembles the products of endogenous and exogenous protease action on receptors for all classes of steroids in a wide range of tissues. The similarities between both the largest and the smallest known forms of the breast tumor estrogen receptor and corresponding forms of other receptors support the notion of the common architecture of steroid receptors in normal and malignant tissues of diverse origins. PMID- 7472274 TI - Species-related differences in steroid-binding specificity of glucocorticoid receptors in lung. AB - The steroid-binding affinities and specificities of pulmonary glucocorticoid receptors have been examined in different mammalian species. The receptors of all species examined are highly specific for binding glucocorticoids, the order of binding affinity being similar to the order of their biological potency. Progesterone also binds significantly to the receptors with an affinity varying between 16% and 41% of the corresponding affinity of corticosterone in different species. The affinity of triamcinolone acetonide for the receptor is highest in the rabbit, lowest in the guinea pig, and intermediate in the rat, mouse, and man, suggesting an inverse relationship between the affinity of glucocorticoids for the receptors and the level of glucocorticoids in blood. Glucocorticoid receptors of cortisol-secreting species (man, guinea pig, rabbit) bind cortisol with a much higher affinity than corticosterone, whereas glucocorticoid receptors of corticosterone-secreting species (rat, mouse) bind corticosterone with a much higher affinity than cortisol. In addition to cortisol, other 17 alpha hydroxylated steroids also bind receptors of cortisol-secreting species with a relatively higher affinity than receptors of corticosterone-secreting species. Thus, in a given species glucocorticoid receptors seem to bind steroids better that are normally secreted by that species than steroids that are not secreted. A change in the pattern of glucocorticoids secreted during development (e.g. the rabbit fetus secretes primarily cortisol whereas the adult rabbit secretes predominantly corticosterone) is not accompanied by a corresponding change in the binding specificity of glucocorticoid receptors. PMID- 7472275 TI - Sex steroids modulate prolactin action in spontaneously luteinizing porcine granulosa cells in vitro. AB - Prolactin (20-1000 ng/ml) stimulated progesterone secretion in monolayer cultures of granulosa cells isolated from mature (greater than 8 mm) porcine follicles. The action of 17 beta-estradiol alone and the interaction between estradiol and prolactin in controlling steroidogenesis were critically dependent upon the duration of hormone treatment. Acute administration (within 36 h) of 17 beta estradiol, but not 17 alpha-estradiol, suppressed both spontaneous and prolactin stimulated progesterone production. Continuation of estrogen treatment beyond 48 h markedly increased progesterone secretion per cell and with prolactin produced synergistic enhancement of progesterone accumulation. However, 5 alpha dihydrotestosterone also significantly augmented progesterone production after both acute and prolonged administration. The steroidogenic effect of prolactin was greater in the presence of 5 alpha-dihydrotestosterone, but did not elicit true synergism. Measurements of sex-steroids levels in porcine follicular fluids indicated that estrogen concentrations that were biologically active in vitro are similar to those attained in vivo. These observations suggest a major role for sex-steroid hormones both in the control of basal steroidogenesis and in the modulation of prolactin action in mature spontaneously luteinizing granulosa cells. PMID- 7472276 TI - A role for dopamine in growth hormone regulation in the dog. AB - The role of dopamine (DA) in the secretion of GH in most animal species is still controversial. We examined in the dog the effect on GH release of nomifensine, a drug which activates both noradrenergic and dopaminergic neurotransmission. Nomifensine (0.4-2.8 mg/kg, iv), administered into 12 unanesthetized male and female beagles, induced short-dose-related rises in canine GH (cGH) levels. Blockade of alpha-adrenergic receptors by phentolamine prevented the GH stimulatory effect of 2.8 mg/kg nomifensine, and an iv bolus injection of haloperidol (a neuroleptic which antagonizes both norepinephrine and DA receptor function) given 45 min before was equally effective. Selective blockade of DA receptors by pimozide significantly reduced the GH-releasing effect of nomifensine. In sum, these data indicated that the effect of nomifensine was the consequence of an enhanced noradrenergic and dopaminergic-neurotransmission. Pretreatment with domperidone, a DA receptor blocker unable to cross the blood brain barrier, failed to modify the GH-releasing effect of nomifensine, suggesting that the DA component subserving the neuroendocrine effect of the drug lies within the blood-brain barrier. Further evidence for a stimulatory role of DA on GH release was the fact that apomorphine, a direct stimulant of DA receptors, induced a rise in cGH levels when administered to dogs pretreated with domperidone. The latter drug was used to prevent emesis and distress due to activation of peripheral DA receptors by apomorphine. However, apomorphine was only active in the dog at doses (250 and 500 microgram/kg, sc) greatly exceeding those active in releasing GH in man, suggesting that the role of DA in cGH regulation is ancillary to that exerted by noradrenergic neurotransmission. In a final study, atropine, a muscarinic cholinergic receptor antagonist, abolished the neuroendocrine effect of nomifensine, a finding which suggests that cholinergic medication plays an important role in cGH regulation. PMID- 7472277 TI - Regulation of ovarian cholesterol metabolism: control of 3-hydroxy-3 methylglutaryl coenzyme A reductase and acyl coenzyme A:cholesterol acyltransferase. AB - Activities of enzymes involved in cholesterol metabolism were measured in ovaries of PMS-hCG-primed immature rats. Microsomal 3-hydroxy-3-methyglutaryl coenzyme A (HMG-CoA) reductase, a rate-limiting enzyme in sterol synthesis, was low during the period of maximal steroid production and cholesteryl ester storage. Acyl CoA:cholesterol acyltransferase (ACAT), the microsomal enzyme catalyzing sterol esterification and cytosolic cholesteryl ester hydrolase and mitochondrial cholesterol side chain cleavage activities, paralleled storage of sterol esters and levels of plasma progesterone, being highest on day 7 post hCG. Further experiments demonstrated that HMG-CoA reductase and ACAT are regulated by different mechanisms than the sterol esterase and cholesterol side chain cleavage system. When blood sterol levels were lowered by 4-aminopyrazolo (3,4 d)pyrimidine, plasma progestin concentrations fell and ovaries failed to accumulate sterol esters. HMG-CoA reductase increased more than 10-fold as a result of this treatment, while ACAT was 15% of that measured in controls. In contrast, cholesteryl esterase and the cholesterol side chain cleavage system were not affected. Intravenous infusion of human lipoproteins reversed the effects of 4-aminopyrazolo (3,4-d)pyrimidine. Raising blood levels up to 6-fold with either cholesterol-supplemented diets or intravenous injections of rat lipoproteins had no effect upon ovarian sterol metabolism, suggesting that the process by which blood cholesterol is utilized is saturated at normal blood sterol levels. Intravenous injection of LH on day 7 post hCG increased HMG-CoA reductase and decreased ACAT activity in addition to depleting cholesteryl esters. Prior treatment of rats with aminoglutethimide prevented the effects of LH on both HMG-CoA reductase and ACAT, indicating that changes in these enzymes were a consequence of the steroidogenic stimulus imposed by LH. Treatment with aminoglutethimide alone on day 7 post hCG did not further depress the already low HMG-CoA reductase activity, but ACAT and cholesterol ester storage were stimulated while sterol esterase activity was not altered. Aminoglutethimide also produced elevated ACAT activity and sterol ester storage in animals treated on day 2 post hCG, a time when there is normally little sterol ester accumulation. We conclude that HMG-CoA reductase and ACAT are regulated, in a reciprocal fashion, by ovarian cholesterol balance. When sufficient cholesterol is available, HMG-CoA reductase is suppressed and ACAT increases to facilitate esterification of sterol in excess of cell needs. When exogenous sterol supplies cannot meet ovarian demands, HMG-CoA reductase rises, and there is a concomitant decline in ACAT. Thus, gonadotropic control over these enzymes may be exerted, in part, through modulation of the supply of cholesterol to the ovary and/or through regulation of the rate of sterol utilization for hormone synthesis. PMID- 7472279 TI - Changes in an electroneutral transport process mediated by angiotensin II in the rat distal colon in vivo. AB - The purpose of these experiments was to determine if angiotensin II mediates the stimulation of water transport in the rat distal colon via an electroneutral mechanism. Amphotericin B (36 micrograms/ml) added to the mucosal surface of the colon significantly increased potential difference (pd) and short circuit current (scc) and significantly decreased resistance. Replacement of the luminal contents of the colon with 150 mM choline chloride decreased pd and scc while resistance remained unchanged. Increased colon resistance occurred on cooling the animals. Thus both increased and decreased pd, scc, and resistance could be measured with the preparation used in this study. The distal colon in vivo is sensitive to angiotensin II since low infusions (7 pmol/kg . min) stimulate water transfer 180% above controls. Transmural pd, resistance, and scc were measured in the distal colon, while the response to the hormone was monitored by measurement of jejunal water transport in the same animals, Intravenous infusion of angiotensin II at a subpressor dose (7 pmol/kg . min) significantly stimulated jejunal water absorption in the absence of any change in transmural pd, resistance, or scc measured in the distal colon. However, infusion of a pressor dose of angiotensin II (700 pmol/kg . min) significantly inhibited jejunal water absorption but again did not alter the electrical properties of the distal colon. These data suggest that angiotensin II can stimulate and inhibit intestinal water transport via electroneutral mechanisms. Thus, the actions of this hormone contrast with that of aldosterone, which stimulates intestinal sodium and water transport by electrogenic mechanisms. PMID- 7472278 TI - Facilitation and inhibition of the estrogen-induced luteinizing hormone surge in the rat by progesterone: effects on cytoplasmic and nuclear estrogen receptors in the hypothalamus-preoptic area, pituitary, and uterus. AB - The possibility that inhibition and facilitation of estrogen-induced LH surges by progesterone could be associated with effects of this steroid on the concentration, subcellular distribution, or molecular properties of estrogen receptors in the hypothalamus-preoptic area (HPOA) and pituitary was investigated. Immature rats (28 days old) that received Silastic capsules containing estradiol-17 beta in oil (150 micrograms/ml) at 0900 h had LH surges between 1700 and 2000 h on day 29. This treatment led to depletion of cytoplasmic estrogen receptors (to 25-35% of control levels) and their accumulation in the nucleus. Insertion of crystalline progesterone implants concomitantly with estradiol implants or 24 h later resulted in blockade or enhancement of the LH surge, respectively. Progesterone administered in either mode did not significantly after the levels of estrogen receptors in the HPOA and pituitary; however, progesterone did suppress the quantity of both cytoplasmic and nuclear estrogen receptors in the uterus when administered in conjunction with estradiol for 24 h oe for 8 h after 24-h estrogen priming. The binding affinity and sedimentation properties in sucrose gradients of cytoplasmic estrogen receptors were unchanged by progesterone treatment. The specificity of the effects of progesterone on LH secretion was examined. Testosterone, dexamethasone, and the synthetic progestin R5020 (17,21-dimethyl-19-nor-4,9-pregnadien-3,20-dione) also inhibited LH surges when injected 8 h after placement of estradiol implants in 28 day-old rats. Only progesterone and R5020 brought about premature and sustained LH release when given to estradiol-primed rats at 0900 h on day 29. None of these compounds interfered directly with the binding of [3H]estradiol to cytoplasmic or nuclear receptors. In conclusion, modulation by progesterone of the estrogen induced LH surge does not seem to result from effects on neural and hypophysial estrogen receptors. In contrast, suppression of uterine estrogen receptors by progesterone may account in part for antagonism by this steroid of estrogen stimulated uterine growth. PMID- 7472280 TI - Role of central nervous system-derived or circulating gamma-aminobutyric acid on prolactin secretion in the rat. AB - To investigate the respective role in PRL secretion of gamma-aminobutyric acid (GABA), either derived from the central nervous system or circulating in plasma, experiments were performed using ethanolamine-O-sulfate (EOS), a specific inhibitor of GABA catabolism. Intracerebroventricular injection of EOS (2 mg/kg) induced in unanesthetized male rats 2-8 h post injection a clear-cut rise in hypothalamic, anterior pituitary (AP), and plasma GABA concentrations. Rises in GABA titers occurred earlier in the hypothalamus and AP (2 h) than in the plasma (4 h). Concomitant to alterations of GABA, there was a striking lowering of plasma PRL evident at 2 h and still present 24 h after EOS administration. In contrast, systemic administration of graded doses of EOS (200-400 mg/kg, iv) did not induce significant changes in plasma GABA concentrations 4 h post injection; only the 600 mg/kg dose of EOS increased GABA concentrations 4 h post injection in the hypothalamo-AP system and decreased plasma PRL concentrations. Finally, in hypophysectomized rats bearing ecotopic pituitaries, despite the occurrence of rises in the hypothalamic GABA after intracerebroventricular or systemic (600 mg/kg) administration of EOS, AP, plasma GABA, and plasma PRL concentrations were not altered. In all these findings indicate that: 1) changes in plasma PRL are best correlated to variations in the amino acid titers occurring in the hypothalamo-AP systems; and 2) circulating GABA does not play a functional role in the control of PRL secretion. Finally, since alterations in blood GABA levels after central or systemic administration of EOS appear to reflect primary changes occurring in the brain concentration of the amino acid, circulating GABA may be a reliable indicator of central nervous system GABAergic function. PMID- 7472281 TI - 17 Alpha-ethinyl estradiol is more potent than estradiol in receptor interactions with isolated hepatic parenchymal cells. AB - Estradiol (E2) and 17 alpha-ethinyl estradiol (EE2) were compared for their abilities to promote nuclear translocation of the hepatic estrogen receptor. High levels of nonradioactive E2 and EE2 were incubated in vitro with isolated rat liver parenchymal cells. Cytosol and nuclear receptors were then partially purified and measured by exchange with [3H]E2. E2 was required in an approximately 100-fold higher concentration than EE2 for promotion of receptor translocation (for example, 10(-5) M E2 was equivalent to 10(-7) M EE2). When the metabolisms of the estrogens by parenchymal cells were compared, it was found that [3H]E2 was metabolized much more extensively than [3H]EE2 after a short time interval. Inhibitors of estrogen metabolism were used to test the hypothesis that the high dose requirement for receptor translocation may be due to metabolism. In isolated liver cells, SKF 525A (beta-diethylaminoethyl-2,2-diphenylpentanoate; an inhibitor of hepatic mixed function oxidase) could increase levels of unmetabolized EE2, cytosol receptor deletion, and nuclear receptor accumulation. SKF 525A alone was not sufficient to increase levels of unmetabolized E2 or receptor translocation in the presence of this estrogen. When testosterone (a transient inhibitor of the 17-oxidoreductase active on E2) was included in cellular incubations (using female liver cells) with E2 and SKF 525A, levels of unmetabolized E2 were substantially increased in conjunction with increased depletion of cytosol receptors and increased levels of nuclear receptors. These data suggest that hepatic metabolism may limit the availability of estrogens for binding to receptors. The present study suggests that E2 is more extensively metabolized than EE2 to metabolites which are less capable of promotion of receptor translocation. PMID- 7472282 TI - Effects of an inhibitor of hepatic drug metabolism, 2-diethylaminoethyl-2,2 diphenylvalerate HCl (SKF 525-A), on thyroxine metabolism in the rat. PMID- 7472283 TI - Evidence for 5 alpha-androstane-3 beta, 17 beta-diol binding to the estrogen receptor in the cytosol from male rat pituitary. AB - The binding of [3H]5 alpha-androstane-3 beta, 17 beta-diol (3 beta-Adiol) in the pituitary cytosol of prepubertal male rats was studied. The following criteria indicated that 3 beta-Adiol was bound to specific proteins and, in all probability, to the estrogen receptor. 1) The binding was of relatively high affinity and low capacity. 2) The complex was destroyed by proteolytic enzymes and by heating. 3) It was precipitable with protamine sulfate and 35% ammonium sulfate. 4) In sucrose linear gradients (5-20%), 3 beta-Adiol was bound in the 7S region. The 3-4S 3 beta-Adiol complex, previously found, was due to the dissociation of 3 beta-Adiol from the 7S complex and further association with nonspecific proteins. 5) Bound hormone was only displaced by 3 beta-Adiol itself and estrogens. 6) Translocation of the estrogen receptor from cytosol to nuclei resulted in a parallel decrease in the number of cytosol 3 beta-Adiol-binding sites. It was also shown that 3 beta-Adiol was not bound to contaminating serum proteins, particularly to alpha-fetoprotein. We interpret these results to indicate that in the male rat pituitary, 3 beta-Adiol is bound to the estrogen receptor. PMID- 7472284 TI - Characterization of long term hormonal cell cultures from the MtT/w15 prolactin- and growth hormone-secreting rat pituitary tumor. AB - PRL-secreting epithelial cell lines (PECL) derived from Furth's MtT/W15 transplantable rat pituitary tumor have been studied as monolayer cultures for up to 18 months. Tumor epithelial cells were mechanically dispersed, plated at high density, and selectively enriched by picking and pooling colonies, thereby avoiding single cell-cloning conditions, in order to study the long term natural evolution of tumor cell populations in vitro. Fibroblast-like cells, prevalent in primary and early passages, underwent a series of morphological changes, which have been interpreted by others as in vitro aging or differentiation, and were eliminated from the cultures within 9 months. PECL phenotypes were heterogeneous and differed from sc MtT/W15 tumors with respect to morphological differentiation and modal chromosome number (48 for solid tumors: 48-92 for PECL; 42 for diploid). PECL were devoid of secretory granules by electron microscopy, but accumulated neutral lipid, complex lysosomes, and endocytotic vesicles. PECL differed from one another in plating efficiency (0.6-7.3%) and basal secretion rates for PRL and GH. Population doubling times decreased during the culture interval from 100 to 24-37 h in different PECL. RIA of medium from early cultures (less than 7 months) disclosed that PRL always exceeded GH levels, mimicking the serum hormone ratio of MtT/W15 tumor-bearing Wistar-Furth rats. Basal hormone release declined with time from a maximal initial rate of 93 micrograms/mg cell protein.24 h for PRL and 2.3 micrograms/mg cell protein.24 h for GH. GH production ceased by 7 months in all PECL, but PRL secretion persisted for 18 months. PRL production increased during exponential growth as a direct function of cell number and protein, and reached a maximum during confluency. Immunoperoxidase-conjugated antibody staining localized PRL in the Golgi region but not in secretory granules, consistent with a rapid release rate for the hormone. In preliminary experiments, rat hypothalamic extract (one hypothalamic extract RP-1 equivalent per dish) and 17 beta-estradiol (1 X 10(-8) M), but not TRH (5 X 10(-8) M), stimulated PRL secretion from PECL/F3, whereas only hypothalamic extract stimulated total cell protein after exposure to the agents for 3-6 days. Long term cultures of PECL exhibited reduced oncogenicity when transplanted in W/Fu rats. During serial passages, in vivo, PRL secretion decreased from 154 microgram/ml serum to the normal range within the same time framework that PRL secretion from cultured PECL/F3 was observed to decrease and stop in vitro. We believe that this phenomenon represents an expressed program within the cells rather than being merely a reflection of the in vitro environment. PMID- 7472285 TI - Actions and interactions of delta 5-androstene-3 beta, 17 beta-diol and 17 beta estradiol in the immature rat uterus. AB - delta 5-Androstene-3 beta, 17 beta-diol (Adiol), in a dosage of 1000 microgram/24 h injected ip into immature female Wistar rats (21-23 days old), induced 72 h after the injection the same elevations of uterine weight, cytosol protein, nuclear DNA, cytosol estrogen and progesterone receptors, and nuclear estrogen receptor levels as did 17 beta-estradiol (E2) in a dosage of 2.5 microgram/24 h. Adiol appeared to translocate the estrogen receptor and induce the synthesis of estrogen and progesterone cytosol receptors in a manner almost identical to that of E2. Studies on the metabolism of tritiated Adiol ruled out the possibility that the estrogenic effects of Adiol might be mediated by conversion to E2. Small doses of Adiol (100 microgram), which alone did not result in estrogenic effects, enhanced the effect of 2.5 microgram E2 on uterine weight and progesterone receptor levels, whereas a dosage of 100 microgram dihydrotestosterone neither enhanced nor inhibited the uterine growth induced by 2.5 microgram E2. A role for the androgen receptor in the synergism between Adiol and E2, therefore, appears to be ruled out. When Adiol in a dosage of 1000 microgram was administered together with 2.5 microgram E2, there was no important difference compared to either steroid alone up to 6 h. Thereafter, a very pronounced and prolonged secondary wave of translocation of the E2 receptor to the nucleus occurred which can explain the synergism between E2 and Adiol that became apparent in the same time interval. This secondary wave of translocation is most likely related to the persistent presence of both Adiol and E2 at the time of replenishment of the cytosol receptor (3-6 h after the injection). It is concluded that delta 5 androstene-3 beta, 17 beta-diol acts in the immature rat uterus like a fully potent estrogen with a complete lack of antiestrogenic effects. In this respect it differs from androgens like testosterone and dihydrotestosterone which can display both estrogenic and antiestrogenic activities. PMID- 7472286 TI - Enzyme-generated intermediates derived from 4-androstene-3,6,17-trione and 1,4,6 androstatriene-3,17-dione cause a time-dependent decrease in human placental aromatase activity. AB - Kinetic evidence is presented for a time-dependent decrease in human placental aromatase activity by enzyme-generated intermediates derived from two widely used steroids previously described as competitive inhibitors of estrogen biosynthesis. Thus, 4-androstene-3,6,17-trione binds to the enzyme with an apparent Ki of 0.43 microM and has a pseudo-first order overall rate constant for decrease in activity of 4.03x10(-3)sec-1, while 1,4,6-androstatriene-3,17-dione has an apparent Ki of 0.18 microM and a pseudo-first order overall rate constant for decrease in activity of 1.10x10(-3)sec-1. These findings imply that the potent inhibition of estrogen biosynthesis caused by these steroids results primarily from a decrease in enzyme activity caused by enzyme-generated intermediates from the parent steroids. PMID- 7472287 TI - Changes in the pituitary-adrenocortical function after intracerebral administration of di-isopropyl-fluorophosphate and physostigmine in the rat. AB - Local administration of di-isopropyl-fluorophosphate (DFP) and physostigmine into the preoptic area produced a marked elevation of plasma corticosterone level in male albino rats. The drugs failed to change the pituitary-adrenocortical function if they were administered into the septum, hippocampus and posterior hypothalamus. The injections were performed through chronically implanted cannulas and the plasma corticosterone level was measured by competitive protein binding assay. Atropine pretreatment inhibited the physostigmine-induced increase of plasma corticosterone level but it did not prevent the changes induced by DFP administration. Dexamethasone pretreatment blocked the plasma corticosterone response level evoked by local application of acetylcholineesterase inhibitors into preoptic area. PMID- 7472288 TI - Studies on the effect of pentobarbiturate, ether and heparin on plasma thyroxine level in rats. AB - In groups of 9 to 20 adult male rats decapitated at 0, 15 or 60 min after i.p. injection of saline or at the same intervals after 2 min exposure to ether no differences in plasma thyroxine level were found. In groups of 5-6 animals a significant decrease of thyroxine level in plasma was observed at 30 min after i.p. injection of pentobarbiturate (40 mg kg-1) and unsignificant decrease after 800 and 2400 U heparin i.p. per animal. Finally, in 58 animals a linear decrease of plasma thyroxine level was found during 480 min beginning from about 60th to 90th min after the onset of pentobarbiturate anaesthesia (40 mg kg-1 followed by 20 mg kg-1 every 60-90 min). It was concluded that the blood clotting preventive dose of heparin (i.e. 300 U per 400 g animal) presumably does not interfere with the level of thyroxine in plasma, while pentobarbiturate apparently results in a considerable decrease of that. PMID- 7472289 TI - Modification of fat and carbohydrate metabolism by neurohypophyseal hormones. I. Effect of lysine-vasopressin on non-esterified fatty acid, glucose, triglyceride and cholesterol levels in the serum of female rats. AB - The effect of the subcutaneous administration of lysine-vasopressin, in doses of 0.625, 1.25, 2.5 and 5 I.U./kg on serum non-esterified fatty acid, triglyceride, cholesterol and glucose levels in fed and fasting female rats was determined. All the doses, in both fed and fasting animals, caused a marked drop in non esterified fatty acid levels about 15 min after starting the experiment. The only exceptions were fed females given doses 2.5 and 5 I.U./kg, in which non esterified fatty acid levels rose. Apart from animals given the smallest dose (0.625 I.U./kg), glucose levels rose during the same interval. In the case of triglyceride levels, a dose of 0.625 I.U./kg produced a decrease and the larger doses (1.25, 2.5 and 5 I.U./kg) an increase in fed females. In fasting animals, triglyceride levels fell (statistically non-significantly) after doses of 0.625, 1.25 and 5 I.U./kg, and rose after 2.5 I.U./kg. The cholesterol level showed a temporary tendency to fall in most of the experiments. PMID- 7472290 TI - Modification of fat and carbohydrate metabolism by neurohypophyseal hormones. II. Effect of lysine-vasopressin on non-esterified fatty acid, glucose, triglyceride and cholesterol levels in the serum of male rats. AB - The effect of subcutaneously injected lysine-vasopressin on serum non-esterified fatty acid, triglyceride, cholesterol and glucose levels in fed and fasting male rats given doses of 2.5 and 5 I.U./kg was determined. About 15 min after starting the experiment, non-esterified fatty acid levels abruptly fell and glucose levels rose in both fed and fasting animals. The only exception were fed males given a dose of 5 I.U./kg in which there was a delayed increase in the non-esterified fatty acid level. Triglyceride levels rose in fed males and fell in fasting males. The cholesterol level fell in both fed and fasting animals. PMID- 7472291 TI - Modification of fat and carbohydrate metabolism by neurohypophyseal hormones. III. Effect of oxytocin on non-esterified fatty acid, glucose, triglyceride and cholesterol levels in rat serum. AB - The authors determined the effect of s.c. injected oxytocin on serum non esterified fatty acid, triglyceride, cholesterol and glucose levels in fed and fasting rats of both sexes, in doses of 2.5 and 5 I.U./kg. No changes in non esterified fatty acid levels were found in fasting animals of either sex. In fed males, non-esterified fatty acid levels rose abruptly after both doses; in fed females they fell after the larger dose. In fed animals of both sexes, the smaller dose of oxytocin produced a decrease, and the larger dose an increase, in triglyceride levels. In fasting females, triglyceride levels fell after both doses; in fasting males a temporary increase was found after the larger dose. In the great majority of cases cholesterol levels displayed a tendency to fall, in both males and females. In the case of glucose, a transient increase was recorded in fed animals of both sexes after the smaller dose and a decrease in males after the larger dose. In fasting animals, glucose levels temporarily rose in every case. PMID- 7472292 TI - Effects of diet, exercise and anorexigenic drugs on serum thyroid hormones. AB - The serum levels of T4, T3 and rT3 were estimated in moderately obese women treated with diet (900-1000 kcal) and 4 h of exercise, supplemented with placebo or anorexigenic drugs (phenmetrazine, phentermine and mazindol). After 12 days of treatment, no change in serum T4 occurred in the placebo group (-4%), while in the groups with anorexigenic drugs a significant rise of T4 was observed (phenmetrazine +33%, phentermine +14% and mazindol +20%). Serum T3 decreased significantly in the placebo and the anorexigenic groups to a similar extent (from -21% to -27%) with the exception of mazindol. We did not notice any decrease of T3 in this group (-3%). A rising trend of serum rT3 was observed in all 4 groups, with a statistical significance for mazindol only. It is suggested that 1) a combined energy deficit produced by diet and exercise leads to a pattern of serum thyroid hormones similar to that observed in resting patients on a total fast; 2) the rise of serum T4 after the administration of anorexigenic drugs is due to their sympathomimetic effects; 3) the largest weight decrease observed in the mazindol group (6.7 kg) may be due to the unchanged serum T3 level, while in the other groups a decrease of serum T3 was observed. The lowest weight loss was observed in the placebo group (4.9 kg) with a pronounced decrease of T3 (-24%). PMID- 7472293 TI - [Effect of reducing diet on the pituitary gland TSH reserve in obesity]. PMID- 7472294 TI - [BLood serum content of various fatty acids in hyperthyroidism]. PMID- 7472295 TI - [Occurrence and possible activity of chromaffin cells in human ovary]. PMID- 7472296 TI - A simple unifying procedure for radioimmunoassay of thyroxine, triiodothyronine and reverse triiodothyronine in unextracted serum. PMID- 7472297 TI - [Isolation and properties of selected pituitary hormones]. PMID- 7472298 TI - Absence of seizures or mirror foci in experimental epilepsy after excision of alumina and astrogliotic scar. AB - In 15 rhesus monkeys (Macaca mulatta) made epileptic by the sensorimotor cortical injection of alumina, the roles of alumina and of "mirror foci" were investigated by serial surgical excisions of the granuloma, surrounding epileptic focus, and contralateral homotopic sensorimotor cortex. Electroencephalographic and electrocorticographic recordings documented foci and transmitted contralateral epileptic activity. After the granuloma was removed, seizures continued but without alumina. After the epileptic cortex was removed, no seizure activity remained and no contralateral independent foci occurred. These findings indicate that the epilepsy incident to alumina injection into the sensorimotor cortex in monkey is not dependent on the continual presence of alumina and is not associated with independent or "mirror foci." PMID- 7472299 TI - Natural history and prognosis of epilepsy: report of a multi-institutional study in Japan. The group for the study of prognosis of epilepsy in Japan. AB - A multi-institutional study on the prognosis of patients with epilepsy was performed in Japan from 1975 to 1977, including 20 institutions. The outcome of epileptic seizures and social adjustment 10, 5, and 3 years after the onset of the illness was studied in three different groups of epileptic patients, respectively. The number of successfully followed-up cases was 1,868 in total, follow-up rate being 42%. The data were analyzed statistically by use of analysis of variance. The rate of remission of seizures in all was 58.3% and the rate of normal social adjustment 62.6%. The remission rate showed no significant difference among the 10-, 5-, and 3-year outcome groups. The study on the outcome of seizure control indicated that the prognosis of seizure control is more favorable (a) in idiopathic than in residual or symptomatic epileptics; (b) in patients with onset before 10 years of age than in those with onset after 10 years; (c) in patients with less frequent seizures; (d) in sleep epilepsy than in waking and diffuse epilepsies; (e) in patients who started treatment within 1 year of onset of seizure; (f) in patients with single-type partial seizures than in those with partial seizures combined with tonic-clonic seizures; (g) in patients without high grade EEG background abnormality; and (h) in patients without neurological symptoms, intellectual deficits, personality disturbances, or psychotic symptoms. Factors that determine the prognosis of social adjustment were almost similar to those for remission of seizures. There was a close correlation between the outcome of seizure control and that of social adjustment. Correlation coefficient between follow-up rate and remission rate in 15 of the 20 collaborated institutions was 0.55 (p less than 0.05), showing a tendency that the remission rate becomes higher as the follow-up rate is raised. PMID- 7472300 TI - Hospitalization rates for epilepsy in the United States, 1973-1976. AB - Data on numbers of admissions and admission rates for all-listed and first-listed epilepsy in nonfederal short-stay hospitals are presented. These statistics are based on 232,000 hospital discharge records from a probability sample of approximately 7,000 hospitals. The patterns of admission rates for epilepsy are compared with the prevalence of epilepsy and trends of admission rates for all conditions. PMID- 7472301 TI - Changes in seizure frequency and test-retest scores on the Wechsler Adult Intelligence Scale. AB - Test-retest performance on the Wechsler Adult Intelligence Scale (WAIS) of two groups of adult epilepsy patients are presented and compared. In one group, Seizures Improved (SI) group, seizure frequency had decreased during the test retest interval, and in the other group, Seizures Unimproved (SU) group, the number of seizures had either increased or stayed the same over the test-retest interval. The SI group showed a significant test-retest improvement on WAIS Verbal IQ, Performance IQ, and Full Scale IQ, as well as on eight of 11 WAIS subtests. In comparison, the SU group showed significant increases only on the Performance IQ and Object Assembly subtest. Furthermore, differences between the two groups were observed in the pattern of test-retest changes seen on the Performance measures relative to the Verbal measures. The results suggest that change in seizure frequency is one of the factors associated with test-retest changes in the intellectual functioning of epilepsy patients. PMID- 7472302 TI - Caffeine- and aminophylline-induced seizures. AB - The epileptogenicity of caffeine and aminophylline was studied in rats. Intraperitoneal administration of caffeine produced an immediate excitation and seizures followed by an encephalopathy. A progression from encephalopathy to seizures was observed following aminophylline administration. During the development of behavioral seizures, the EEGs showed a progression from fast activity isolated or bursts of spikes and/or sharp waves to full-blown epileptiform discharges. A wide spectrum of cardiac arrhythmias was observed during and following the seizures. Epileptiform activities ranging from isolated or bursts of spikes and/or sharp waves to continuous epileptiform discharges were observed following local application of the drugs to the cerebral cortex. PMID- 7472303 TI - Power spectral analysis of EEG characteristics during sleep in epileptics. AB - Power spectral analysis of spontaneous EEG activity recorded during non-rapid eye movement (REM) sleep was employed in an effort to identify characteristic EEG substrate abnormalities in epileptics. All-night polygraphic recordings were obtained from 10 poorly controlled subjects with mixed and generalized seizures. Similar data were obtained from 10 matched nonepileptic subjects. Continuous 10 min samples of non-REM sleep were identified at the beginning and end of these all-night recordings. EEG activity recorded during these periods from left sensorimotor and parieto-occipital bipolar leads was digitized, subjected to the fast Fourier transform, and then sorted into successive 4 Hz frequency bands between 0 and 23 Hz. Mean spectral density was determined for each frequency band from both samples in both groups. Comparison of these values disclosed both similarities and differences between the two groups. All subjects showed a significant "time-of-night effect," with power at 0-3, 4-7, and 8-11 Hz greater at the beginning of the night. This finding was related to a similar observation in developing infants and attributed to a circadian influence on neuronal generators. In group comparisons, epileptics showed selective differences in common when compared with nonepileptics. These differences were reduced when data from a small group of unmedicated epileptic subjects were partialled out. Medicated epileptics showed differences restricted to the early morning non-REM sleep sample, which included significantly greater power at 4-7 Hz and significantly lower power at 12-15 Hz. Unmedicated subjects showed these differences in both samples and also showed significantly greater power in the 0 3 Hz band. It was suggested that anticonvulsant medications may exert a general normalizing influence on EEG characteristics during sleep, particularly at the beginning of the night in proximity to their administration. The documentation of significant EEG deviations restricted to two basic rhythmic patterns during sleep suggests a disturbance of underlying thalamocortical mechanisms. PMID- 7472304 TI - Deficits in neuropsychological functioning and psychopathology in persons with epilepsy: a rejected hypothesis revisited. AB - Previous investigations have failed to detect a relationship between the adequacy/inadequacy of neuropsychological functioning and psychopathology in persons with epilepsy. It is argued that these results were obtained because of the manner in which the measures of psychopathology, the MMPI, were analyzed. The MMPI data from two recent neuropsychological investigations of epilepsy were analyzed using a technique which has been found to be quite sensitive to various psychopathological categories, i.e., Goldberg's Rules. Comparison of subjects varying in degree of neuropsychological impairment revealed increased psychopathology in the patient groups with more widespread cognitive deficits. It is concluded that there is a relationship between deficits in neuropsychological functioning and psychopathology in epilepsy. This relationship is related to the larger literature of epilepsy and psychopathology. PMID- 7472305 TI - Diphenylhydantoin and primidone in tears. AB - Diphenylhydantoin (PHT) and primidone (PRM) were determined by the EMIT technique in the plasma, tears, saliva, and CSF of epileptic patients. Results indicate for PHT that tear values are more strictly correlated than are the saliva values to plasma and CSF concentrations. As for PRM, the data obtained show great interindividual variability of concentration in the different body fluids--in agreement with the wide range of both protein binding and half-life of this drug. PMID- 7472306 TI - An immunological model of epilepsy: seizures induced by antibodies to GM1 ganglioside. AB - Following observations that the intracerebral injection into rats of antiserum to brain gangliosides resulted in recurrent epileptiform activity and that seizure activity was not seen if antibodies were removed by absorption of the antiserum with pure GM1 ganglioside, a study was undertaken to establish characteristics of the immunological agents used to produce this model of epilepsy. It was determined that the potencies (antibody titers with GM1 ganglioside) of antiganglioside sera can be correlated with the intensities of epileptiform activity they induce; that immunoglobulin fractions from antiganglioside sera are even more effective biologically than the antisera; and that antibodies to GM1 ganglioside purified by affinity chromatography can also induce recurrent epileptiform discharges but are not as effective as either native antiserum or immunoglobulin fractions. PMID- 7472307 TI - Effects of age of onset of tonic-clonic seizures on neuropsychological performance in children. AB - Forty-eight children (aged 9 to 15 years) with tonic-clonic seizures were administered a neuropsychological test battery. The children with seizures of early onset (before age 5) were significantly impaired relative to the children with later onset on 8 of the 14 measures in the battery. The deficits were seen on tasks whose requirements included the repetition of a simple motor act, attention and concentration, memory, and complex problem solving. These findings emphasize the need for further research to determine the causal factors of the greater dysfunction seen in the early onset group. PMID- 7472308 TI - Effects of diazepam on hippocampal excitability in the rat: action in the dentate area. AB - Acute and chronic experiments were performed on rats to examine the effects of diazepam (Valium) on recurrent inhibition in the monosynaptic perforant path dentate synapse of the dentate area of the dorsal hippocampus. Evidence was obtained which indicated that diazepam facilitated a presumably GABA mediated post synaptic recurrent inhibition in both acute and chronic preparations at 1 and 2 mg/kg doses (IP). Acute studies also revealed that diazepam prolonged recurrent inhibition, possibly by lengthening the IPSP. An effect of the drug on cholinergically mediated positive feedback from the septum could not be ruled out, however. Taken together, this study extends the evidence that diazepam acts centrally on GABA mediated inhibition in the limbic system. Furthermore, the limbic action of diazepam revealed here suggests a mechanism for the anticonvulsant properties of diazepam in epilepsy involving subcortical (limbic) circuitry. PMID- 7472309 TI - Asbestos distribution in the Central Serpentine District of Maryland Pennsylvania. PMID- 7472310 TI - Phagocytosis by alveolar cells studied by electron microscopy and microanalysis: comparison of two types of particles. PMID- 7472311 TI - A comparison of skull and femur lead levels in adult rats. PMID- 7472312 TI - Primary pericardial mesothelioma following exposure to asbestos. PMID- 7472313 TI - Asbestosis in a Barcelona fibrocement factory. PMID- 7472314 TI - Factors influencing the transport of lead across the small intestine of the rat. PMID- 7472315 TI - Development of glycine- and GABA-gated currents in rat spinal motoneurons. AB - 1. Developmental changes in glycine- and gamma-aminobutyric acid (GABA)-activated currents were studied in spinal motoneurons of embryonic and neonatal rats with the use of whole cell recording techniques. 2. Pressure ejection of glycine or GABA onto motoneuron somata produced Cl(-)-mediated inward currents and membrane depolarizations. During embryonic development, the average amplitude of GABA gated currents was threefold larger than that of glycine-gated currents, but as a result of a large eightfold postnatal increase in glycine-activated currents, similar currents were produced by both amino acids after birth. 3. At all ages the decay of glycine- and GABA-gated currents best fit one-exponential curve, and their time constants were similar. The average decay time constant decreased by twofold after birth. 4. The ionic specificity of glycine- and GABA-gated channels was studied to determine whether the large amplitude of GABA-activated currents in embryonic motoneurons resulted from the contribution of an outward HCO-3 movement. Manipulations of Cl- and HCO-3 concentrations produced changes in the reversal potentials of glycine and GABA that were similar to the calculated changes in the equilibrium potentials of Cl-. This suggested that glycine- and GABA-gated currents were Cl- specific, and HCO-3 movement did not contribute more to the current generated by GABA than that produced by glycine. 5. Glycine- and GABA-gated currents were associated with severalfold increases in membrane conductance. The conductance increase generated by GABA in embryonic motoneurons was sevenfold larger than that generated by glycine, but similar conductance changes were produced by both amino acids after birth.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7472316 TI - Effects of Ca2+ channel blockers on directional selectivity of rabbit retinal ganglion cells. AB - 1. Extracellular recordings were made from ON-OFF directionally selective ganglion cells in superfused rabbit retinas in order to examine the effects of voltage-activated Ca2+ channel blockers on the response of these ganglion cells to a moving bar of light. 2. Bath application of Cd2+ (67-110 microM) abolished directional selectivity in the ganglion cells. That is, the cells gave nearly equal responses to the leading and trailing edges of a bar of light moved in the preferred and null directions. This effect of Cd2+ was rapidly reversible. 3. Directional selectivity in the ganglion cells was not affected by Ni2+ (120-440 microM), Co2+ (180-690 microM), or the L-type Ca2+ channel blockers nicardipine (7-29 microM) and methoxyverapamil (18-60 microM). These blockers did, however, reduce the responses of the ganglion cells to a bar of light moved in the preferred direction. 4. omega-Conotoxin MVIIC (130 nM-1.9 microM), which potently blocks N-type and Q-type Ca2+ channels, abolished directional selectivity in the ganglion cells. omega-Conotoxin MVIIC not only brought out large leading and trailing edge responses to movement of a bar of light in the null direction, but it also increased the leading and trailing edge responses to movement of the bar of light in the preferred direction. The effect of omega-conotoxin MVIIC was slowly reversible. 5. The N-type Ca2+ channel blocker omega-conotoxin GVIA (1.4 6.3 microM) did not abolish directional selectivity in the ganglion cells. This blocker did, however, bring out some response to the leading edge of a bar of a light moved in the null direction. This effect of omega-conotoxin GVIA appeared to be irreversible. 6. omega-Agatoxin IVA, a potent blocker of P-type Ca2+ channels, when bath applied at low concentrations (66-83 nM), increased the responses to movement of a bar of light in the preferred direction but brought out only small responses to movement of the bar of light in the null direction. At high concentrations (250-280 nM) that reportedly block Q-type Ca2+ channels by > or = 50%, omega-agatoxin IVA nearly abolished directional selectivity. This effect of omega-agatoxin IVA was slowly reversible. 7. These results indicate that omega-conotoxin MVIIC- and omega-agatoxin IVA-sensitive Ca2+ channels (possibly Q-type channels) play an important role in the generation of directional selectivity in rabbit retinal ganglion cells. PMID- 7472317 TI - Roles of specific metabotropic glutamate receptor subtypes in regulation of hippocampal CA1 pyramidal cell excitability. AB - 1. Metabotropic glutamate receptors (mGluRs) are coupled to various second messenger systems through guanosine 5'-triphosphate-binding proteins. To date, at least seven mGluRs have been cloned, and these mGluR subtypes can be divided into three major groups on the basis of similarities in amino acid sequence, coupling to second-messenger cascades in expression systems, and pharmacological profiles. These groups include group I (mGluR1 and mGluR5), group II (mGluR2 and mGluR3), and group III (mGluR4, mGluR6, and mGluR7). 2. On the basis of its selective activation of phosphoinositide hydrolysis in brain slices and its ability to activate mGluR1a expressed in Xenopus oocytes, others have suggested that 3.5 dihydroxyphenylglycine (DHPG) may be selective for group I mGluRs. Consistent with this hypothesis, we report that DHPG also activates mGluR5 expressed in oocytes, whereas it is inactive at mGluR4 and mGluR7 expressed in baby hamster kidney (BHK) cells. The compound (2S,1'R,2'R,3'R)-2-(2.3 dicarboxycyclopropyl)glycine (DCG-IV) activates both mGluR2 and mGluR3 at submicromolar concentrations, whereas it is inactive at mGluR4 and mGluR1, suggesting that this compound may be selective for group II mGluRs. Consistent with this hypothesis, we find that DCG-IV does not activate mGluR5 expressed in oocytes and does not activate mGluR7 expressed in BHK cells. These findings suggest that DHPG and DCG-IV are highly selective agonists for group I and group II mGluRs, respectively. 3. Previous studies that have examined the physiological roles of mGluRs have generally used agonists that do not differentiate between the various subtypes.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7472318 TI - Functional linkages between motor cortical cells and elbow flexor muscles. Evidence for and characteristics of postspike facilitation. AB - 1. Two monkeys (Macaca fascicularis) making high-level but submaximal isometric flexions of the elbow were investigated for the output effect of motor cortical cells on the electromyogram (EMG) activity of two main elbow flexors using the method of spike-triggered averaging of rectified EMGs (STAs). 2. Monkeys were trained to perform individual isometric contractions for > 2 s, and two series of > or = 20 contractions, the second series being at a greater force. EMG electrodes pairs were implanted in the biceps brachii and brachioradialis. A total of 257 cortical cells were found that discharged with the active and passive movements of the elbow. We examined the EMG postspike facilitations (PSFs) produced in either one or the two flexors for only those cells that discharged during the isometric contraction, and provoked PSFs in the two series of contractions. 3. The main characteristics of the EMG isometric contractions in the agonists were analyzed. Spectral analysis showed that the increases in the EMG median frequency with force stabilized at the force levels performed by monkeys. Cross correlation methods showed no cross talk between agonists. 4. The 26 selected cortical cells had a regular discharge frequency. Ten cells did not change frequency with a 22-30% force increase, 14 cells discharged at a higher frequency, and 2 cells discharged at a lower frequency. For single-cell frequencies of 5-65 Hz, interspike intervals < 10 ms were rare: the median and modal intervals were 20-30 ms. 5. The significance of PSFs with respect to the EMG background noise was estimated statistically. STAs from successive epochs under identical load conditions, and STAs performed at a distance from the trigger, showed that PSFs were authentic postspike effects and not sudden EMG changes synchronized by chance with the triggering cell. The features distinguishing PSF from secondary postspike EMG changes or coactivation and task related effects were studied in simultaneous STAs of flexors and autocorrelogram of cortical spikes. 6. The magnitude of the PSF was expressed as the percent peak amplitude above the mean EMG baseline. The mean percent amplitude of the 90 PSFs produced in both muscles and series was 4.0 +/- 2.4% (mean +/- SD). There was no difference in the average amplitude of PSFs in the two flexors, although the baseline voltages in the biceps brachii were higher. Neither was there any significant change with force while the baseline level increased by 29 +/- 10%, indicating that the absolute PSF amplitude increased in the same proportion as baseline.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 7472319 TI - Directionally selective motion detection in the sustaining fibers of the crayfish optic nerve: linear and nonlinear mechanisms. AB - 1. Directional selectivity of crayfish sustaining fibers was examined with drifting sine wave gratings and with intracellular and extracellular recordings. Directionality was measured for variations in stimulus contrast, spatial frequency, and temporal frequency. 2. Sustaining fibers exhibit directional selectivity in the magnitude of the compound postsynaptic potential (PSP), the impulse frequency modulation response, and the mean firing rate. The mean synaptic potential is insensitive to direction. The directionality of the mean impulse rate appears to arise by rectification in the voltage-to-impulse transduction. 3. The preferred directions of three identified sustaining fibers are similar to those of head-down optomotor neurons to which these sustaining fibers project. 4. The modulatory response, elicited by gratings drifting in the preferred direction, increased linearly with contrast until saturation (typically at a contrast of 0.5), where maximum directional selectivity obtains. 5. The magnitude of the directional response is a band-pass function of spatial and temporal frequency and exhibits reversal of directionality (i.e., aliasing) at high spatial and temporal frequencies. The results imply a spatial sampling interval of 4.5 degrees and a temperature-dependent inhibitory delay of 40-90 ms. The PSP modulation response shares several features with that of neighboring tangential (Tan1) neurons. 6. A qualitative model is proposed for the transformation of a phase-sensitive, linear directional response to a phase insensitive and nonlinear time-averaged response, based on the functional connections from Tan1 neurons to sustaining fibers to optomotor neurons. The model includes a threshold rectification, a synaptic band-pass filter, and differences in temporal phase among converging modulatory signals. PMID- 7472320 TI - Kinetic properties of NMDA receptors in embryonic Xenopus spinal neurons. AB - 1. The kinetic properties of embryonic Xenopus spinal cord neuron N-methyl-D aspartate (NMDA)-activated receptors (NMDAR) were examined at the single-channel level. These receptors have a main conductance state of 50 pS and make occasional sojourns to a subconductance level of approximately 40 pS. The open channel lifetime is 1.3 ms at -80 mV and in 1 mM Ca(2+)-, Mg(2+)-free solution. Extracellular Mg2+ blocks the channel at a rate of 2.9 x 10(8) M-1 s-1 and with a Kd of approximately 20 microM at -80 mV. In patches with only one channel active, the closed interval duration distribution requires at least four exponentials to be fit. The time constant of one closed interval component decreases with increasing NMDA concentration. Kinetic modeling indicates that these NMDA receptors open at a rate of 230 s-1 and close at a rate of 167 s-1, that in the absence of desensitization the maximum probability of being open is approximately 0.5, and that this probability is half-maximal at approximately 150 microM NMDA. PMID- 7472321 TI - Mnemonic firing of neurons in the monkey temporal pole during a visual recognition memory task. AB - 1. We examined single-neuronal activity in the temporal pole of monkeys, including the anterior ventromedial temporal (VMT) cortex (the temporopolar cortex, area 36, area 35, and the entorhinal cortex) and the anterior inferotemporal (IT) cortex, during a visual recognition memory task. In the task, a trial began when the monkey pressed a lever. After a waiting period, a visual sample stimulus (S) was presented one to four times on a monitor with an interstimulus delay. Thereafter, a new stimulus (R) was presented. The monkeys were trained to remember S during the delay period and to release the lever in response to R. Colored photographs of natural objects were used as visual stimuli. 2. About 70% of the recorded neurons (225 of 311) responded to at least one of the Ss tested. Thirty percent of these neurons (68 of 225) continued to fire during the subsequent delay periods. In 75% of these neurons (51 of 68), the firing during the delay period strongly correlated with the response to S. 3. The discharge rate during the delay period did not correlate with the monkey's eye movements, pressing or releasing of the lever, or the reaction time. 4. If the monkey erroneously released the lever in response to S or during the delay period, the firing disappeared after the erroneous lever release. If the monkey failed to release the lever in response to R, the firing persisted even after R was withdrawn. The discharge rate in incorrect trials was comparable with that in correct trials. The neurons were considered to fire for as long as the memory of S was necessary. 5. Firing persisted even when an achromatic version or half (even a portion) of S was presented, indicating that the color, a particular portion, or the entire shape of S was not always necessary to elicit firing. 6. An S that elicited firing during the delay period invariably elicited a visual response. Neurons that fired during the delay period showed a higher stimulus selectivity than other visually responsive neurons in the anterior VMT cortex. Thus neurons that fire during the delay period represent a subgroup of visually responsive neurons that are selectively tuned to a certain stimulus. 7. More neurons fired during the delay period in the anterior VMT cortex than in the anterior IT cortex. 8. We conclude that firing during the delay period by neurons in the temporal pole reflects the short-term storage of visual information regarding a particular S. PMID- 7472322 TI - Electrophysiological evidence that dentate hilar mossy cells are excitatory and innervate both granule cells and interneurons. AB - 1. The hypothesis that dentate hilar "mossy" cells are excitatory was tested by simultaneous intracellular recording in rat hippocampal slices. Mossy cells were recorded simultaneously with their potential targets, granule cells and interneurons. The gamma-amino-butyric acid-A (GABAA) receptor antagonist bicuculline was used in most experiments to block the normally strong inhibitory inputs to granule cells that could mask excitatory effects of mossy cells. Some cells were recorded with electrodes containing the marker Neurobiotin so that their identity could be confirmed morphologically. 2. A mossy cell action potential was immediately followed by a brief depolarization in a granule cell in 20 of 1,316 pairs (1.5%) that were recorded in the presence of bicuculline. The mean amplitude of depolarizations was 1.99 +/- 0.24 (SE) mV when the postsynaptic membrane potential was -55 to -65 mV. Depolarizations could trigger an action potential if the granule cell was depolarized from its resting potential so that its membrane potential was -50 to -60 mV. These data suggest that mossy cells excite granule cells monosynaptically. 3. Monosynaptic excitation of an interneuron by a mossy cell was recorded in 4 of 47 (8.5%) simultaneously recorded mossy cells and interneurons, also in the presence of bicuculline. The mean interneuron depolarization was 1.64 +/- 0.29 mV when the interneuron membrane potential was approximately -60 mV. When an interneuron was at its resting potential (-52 to -63 mV), action potentials were often triggered by the depolarizations. 4. Without bicuculline present, mossy cells had no apparent monosynaptic effects on granule cells, as has been previously reported. However, effects that appeared to be polysynaptic were observed in 5 of 92 pairs (5.4%). Specifically, a small, brief hyperpolarization occurred in granule cells 2.5-7.3 ms after the peak of a mossy cell action potential. Given the results indicating that mossy cells excite interneurons, and the long latency to onset of the hyperpolarization, one possible explanation for the hyperpolarization is that mossy cells excited interneurons that inhibited granule cells. 5. The results suggest that mossy cells are excitatory neurons. In addition, mossy cells appear to innervate both granule cells and interneurons that are located within several hundred micrometers of the mossy cell soma. The only detectable effect on granule cells in this area under normal conditions appears to be disynaptic and inhibitory. However, when GABAA-receptor-mediated inhibition is blocked, monosynaptic excitation of granule cells by mossy cells can be detected. PMID- 7472323 TI - Acetylcholine receptor desensitization induced by nicotine in rat medial habenula neurons. AB - 1. The activation and desensitization properties of nicotinic acetylcholine receptor (nAChR) channels were examined in acutely isolated medial habenula (MHb) neurons using whole cell patch-clamp recordings. nAChR-mediated currents were evoked by applying known concentrations of nicotinic agonists using rapid solution exchange techniques. 2. At a membrane potential of -60 mV, nAChR currents were observed above a concentration of approximately 100 mM nicotine. The peak current amplitude at low doses of agonist was proportional to the square of the concentration of nicotine, indicating that at least two molecules of agonist were required for channel opening. The concentration of nicotine required for half-maximal nAChR activation was estimated as 77 microM from a complete concentration-response curve. 3. During the continuous activation (2-5 s) of nAChRs by high concentrations of nicotine (300 microM), the current desensitized rapidly and extensively. The desensitization phase was described by the sum of two exponentials, with time constants of 210 and 1,435 ms. The fast component comprised 74% of the desensitizing phase of the current. Recovery from desensitization induced by 2- s applications of 300 microM nicotine was also fast and could be reasonably well described by a single exponential with a time constant of approximately 800 ms. Both the time courses of desensitization and recovery from desensitization were slightly slower at positive membrane potentials. 4. Incubation of neurons with low concentrations of nicotine (100 nM 10 microM) caused a slowly developing but pronounced desensitization of the nAChRs. In these cases desensitization was assessed from the reduction in the amplitude of the peak nicotinic current induced by repetitively applied pulses of a higher test concentration of agonist. A 5-min continuous exposure to 1 microM nicotine reduced the amplitude of the acetylcholine (30 microM, 1 s) test response to < 30% of its control value. As with higher concentrations of nicotine, the onset of the desensitization induced by 1 microM nicotine was biexponential, with fast and slow time constants of 15 s and 1.74 min, respectively. Recovery from the desensitization induced by these longer applications of nicotine was much slower than that observed with the brief pulses of high concentrations of nicotine. The concentration required for half-maximal desensitization after a 5-min incubation was approximately 300 nM. 5. Peak nAChR currents were approximately 85% smaller at +40 mV compared with -40 mV. The receptors that do not open at positive potentials desensitize almost as well as they would at negative potentials after channel opening.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 7472324 TI - Electrophysiological and pharmacological characterization of a mammalian Shaw channel expressed in NIH 3T3 fibroblasts. AB - 1. The Shaw-like voltage-activated potassium channel Kv3.1 is expressed in neurons that generate rapid trains of action potentials. By expressing this channel in a mammalian cell line and by simulating its activation, we tested the potential role of this channel in action potential repolarization. 2. NIH 3T3 fibroblasts were stably transfected with Kv3.1 DNA. Currents recorded in these cells had a threshold of activation at approximately -10 mV, showed little inactivation, and were very sensitive to blockade by 4-aminopyridine and tetraethylammonium. 3. Kv3.1 currents activated rapidly at the onset of depolarizing voltage pulses. After an initial rapid phase of activation, which could be fit by an n4 Hodgkin-Huxley model, Kv3.1 currents expressed in fibroblasts had a second, slower phase of activation, and, in some cells, a slower phase of partial inactivation, both of which could be fit with modified n4p models. 4. Cell-attached single-channel recordings indicated that the Kv3.1 channel displays two gating behaviors, a short-open-time pattern, which occurs only at the onset of depolarization, and a long-open-time pattern, which predominates during prolonged depolarizations. 5. The amplitude of Kv3.1 currents, and the probability of channel openings, was reduced by a phorbol ester activator of protein kinase C, and the action of this agent was blocked by preincubation with the protein kinase inhibitor H7 (1-[5-isoquinolinesulfonyl]-2 methyl piperazine). In contrast, the effects of dioctanoyl glycerol, which also attenuated the currents, could not be completely blocked by H7, suggesting that diacylglycerols may act on the channel by a kinase-independent pathway. 6. Incorporation of a current with the kinetics and voltage dependence of Kv3.1 currents into a model cell with a sustained inward current showed that, in contrast to other delayed-rectifier currents such as the Shaker-like Kv1.1 and Kv1.6 channels, the level of expression of Kv3.1 currents could be varied over a wide range without attenuation of action potential height. Our results suggest that the Kv3.1 channel may provide rapidly firing neurons with a high safety factor for impulse propagation. PMID- 7472325 TI - Effects of a spider toxin and its analogue on glutamate-activated currents in the hippocampal CA1 neuron after ischemia. AB - 1. We studied the effects of polyamine toxins derived from a spider venom on CA1 pyramidal neurons in gerbil hippocampal slices by patch-clamp recording. Joro spider toxin (JSTX) and its synthetic analogue, 1-naphthyl acetyl spermine (Naspm), which are known to block non-N-methyl-D-aspartate (non-NMDA) receptor in a subunit specific manner, were used. 2. Naspm depressed the excitatory postsynaptic currents (EPSCs) mediated by non-NMDA receptor channels. A further reduction of EPSCs occurred with addition of 6-cyano-7-nitroquin-oxaline-2,3- dione (CNQX). Conversely, when CNQX was applied first, no further depression of EPSCs occurred on addition of Naspm, indicating that Naspm blocks a fraction of the CNQX-sensitive non-NMDA-receptor-mediated currents. 3. After ischemia, the time course of EPSCs of CA1 pyramidal neurons was slowed and Naspm depressed the slow EPSCs more strongly than those in control neurons. 4. Analysis of single channel currents by outside-out patch-clamp recording from ischemic CA1 neurons revealed that Naspm blocked a subpopulation of alpha-amino-3-hydroxy-5 methylisoxazole-4-propionate- and kainate-induced single-channel currents. 5. Because the EPSCs in CA1 neurons after ischemia are mediated by Ca(2+)-permeable non-NMDA receptor-mediated conductances, the present results indicate that Naspm and JSTX are effective at blocking abnormal EPSCs that may induce Ca2+ accumulation leading to delayed neuronal death after transient ischemic insult. PMID- 7472326 TI - Kinematics of spontaneous, reflex, and conditioned eyelid movements in the alert cat. AB - 1. Upper eyelid position and velocity, and the electromyographic (EMG) activity of the orbicularis oculi muscle, were recorded bilaterally in alert cats during spontaneous, reflexively evoked, and conditioned eyelid movements. 2. Spontaneous blinks appeared randomly (0.2-0.5 per min) and consisted of a fast, large downward lid movement followed by a slower up phase. Blinks of smaller amplitude and slower velocity were also observed mainly accompanying behavioral movements, such as during peering and grimacing. 3. Eyelid response to air puffs applied to the cornea and tarsal lid skin consisted of a short-latency (9-16 ms), fast (up to 2,000 degrees/s) downward movement that lasted for 25-30 ms, followed by late, small downward sags that were sometimes still evident after stimulus offset. Blinks outlasted the duration of the stimulus by approximately 150 ms. Blinks elicited by flashes of light or tones showed longer latency (47.3 +/- 6.3 and 53.7 +/- 8.0 ms, mean +/- SD; respectively), smaller amplitude, and a quicker habituation than air-puff-evoked lid responses. 4. For the down phase of the blink, the peak velocity, but not its duration, increased linearly with blink amplitude. Because the rise time of the down phase remained constant, changes in blink amplitude seemed to be the result of increased blink velocity. The down phase of a typical 10 degrees blink was 10 times faster than the up phase of the same blink or than upward and downward lid saccades of the same amplitude. The peak velocity and duration of the up phases of reflex blinks and upward and downward lid saccades increased linearly with lid movement amplitude. 5. The initial down phase of air-puff-evoked blinks decreased in latency, increased in amplitude and peak velocity, and maintained the same rise time for increasing puff pressure. None of these parameters was dependent on puff duration. The duration of the blink also increased linearly with air puff duration. 6. The amplitude of air-puff-evoked blinks was inversely related to lid position, decreasing with further lid positions in the closing direction. In contrast, neither peak nor integrated EMG activity of the orbicularis oculi muscle was affected by lid position, being only a function of stimulus parameters and of the animal's level of alertness. 7. Air puffs > 20 ms and > 1 kg/cm2 evoked two successive bursts (R(ap) 1 and R(ap) 2) in the EMG activity of the orbicularis oculi muscle. Shorter and/or weaker stimuli evoked only the R(ap) 1 response.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 7472327 TI - Properties of transmission at a giant glutamatergic synapse in cerebellum: the mossy fiber-unipolar brush cell synapse. AB - 1. The synaptic activation by mossy fibers (MFs) of unipolar brush cells (UBCs) in the vestibular cerebellum (nodulus and uvula) was examined using patch-clamp recording methods in thin, rat cerebellar slices with Lucifer yellow-filled pipettes for subsequent fluorescence microscopic verification of the cell morphology. 2. UBCs were distinguished from adjacent granule cells in thin cerebellar slices in the uvula and nodulus regions by their larger soma diameters and short dendritic brush, greater whole-cell capacitance, and a prolonged, biphasic excitatory postsynaptic current (EPSC) to stimulation of MFs. 3. Thin section transmission electron micrographs of the MF-UBC synapse displayed an unusually extensive area of synaptic apposition estimated to measure 12-40 microns2. The majority of UBCs was innervated by a single MF. At high magnification, individual clusters of presynaptic vesicles could be discerned, separated by regions of presynaptic membrane lacking vesicles, but apposed to continuous regions of postsynaptic density. Thus, after release, transmitter diffusion from the synaptic cleft must traverse considerable stretches of postsynaptic membrane before escape into extracellular space. In contrast, MF granule cell synapses in these cerebellar regions resembled glutamate synapses in other brain regions in that the total synaptic area measured < or = 4 microns2. These synaptic junctions were flanked by short stretches of unspecialized plasma membrane, providing a short (0.5 micron) diffusional path from the site of neurotransmitter release to a branch point of the extracellular space. 4. The MF evoked EPSC in UBCs was composed of a fast (10-90% rise time: 0.70 ms) and slow (10-90% rise time: 395 ms; 10-90% decay time: 3.1 s) component. The fast component was blocked by the alpha-amino-3-hydroxy-5-methyl-4-isoxazolepropionic acid/kainate (AMPA/KA) antagonist 6-cyano-7-nitroquinoxaline-2,3-dione (10 microM) and displayed linear current-voltage (I-V) relations in the presence or absence of external magnesium. 5. The slow EPSC was also mediated by glutamate receptors, but in most neurons both AMPA/KA and N-methyl-D-aspartate (NMDA) receptors contributed to the slow EPSC, with the contribution of NMDA receptors predominating in the majority of cells. Consequently, although all cells displayed linear I-V relations in Mg(2+)-free saline, cells in which the slow EPSC was predominently mediated by NMDA receptors exhibited voltage-dependent rectification in the presence of external Mg2+ (1 mM). 6. With increasing postnatal age (10-30 d), the contribution made to the slow EPSC by NMDA receptors declined, with a reciprocal increase in the contribution being made by AMPA/KA receptors.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 7472328 TI - Taste responses in neurons in the nucleus of the solitary tract that do and do not project to the parabrachial pons. AB - 1. Mechanisms of neural coding of gustatory stimuli were studied in the nucleus of the solitary tract (NTS), the first relay in the neural pathway for gustation, in anesthetized rats. Taste-responsive NTS units were identified as "relay" or "nonrelay" based on the electrophysiological response to electrical pulses delivered to the parabrachial nucleus of the pons (PbN), the second relay in the neural pathway for gustation. Coding mechanisms in each group were analyzed separately. 2. Taste responses to sapid solutions of NaCl (0.1 M), HCl (0.01 M), quinine HCl (0.01 M), sucrose (0.5 M) and Na-saccharin (0.004 M) were recorded in single units in the NTS. After gustatory stimulation, electrophysiological responses to electrical stimulation of the taste-responsive part of the ipsilateral PbN were recorded. A 0.2-ms pulse was delivered at 75-250 microA at a rates of 1, 25, 50 and 100 pps through a bipolar stainless steel electrode. An antidromic response was defined as a time-locked spike that occurred at a fixed latency after PbN stimulation that followed high stimulation frequencies. A collision test also was performed. 3. Of 42 taste-responsive NTS units, 19 (45%) were relay units, 22 (52%) were nonrelay and 1 unit was activated orthodromically by PbN stimulation. Latencies of evoked spikes ranged from 1.75 to 4.0 ms 2.1 +/- 0.2 ms (mean +/- SE, median, 1.75 ms). 4. Examination of general response characteristics revealed few differences among relay and nonrelay units.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7472329 TI - Corticofugal influence on taste responses in the nucleus of the solitary tract in the rat. AB - 1. Previous work has revealed a pervasive influence of the gustatory neocortex (GN) on the electrophysiological responses to taste in the parabrachial nucleus of the pons (PbN), the second synapse in the central pathway for gustation. Subsequent experiments have further suggested that direct projections from the GN to the PbN are not sufficiently dense to account for the widespread effects of cortical input. Because the main source of input to the PbN, i.e., the nucleus of the solitary tract (NTS), also receives input from the GN, the present experiment was conducted to test the hypothesis that changes in taste responses in the PbN after temporary elimination of GN input may be a normal reaction to altered input originating in the NTS. 2. Fourty-three taste-responsive neurons in the NTS were isolated initially in urethan-anesthetized rats. Single units were then classified as "relay" (n = 12) or "nonrelay" (n = 13) on the basis of their electrophysiological response to electrical shocks delivered to the taste responsive portion of the PbN. After histological analyses, 18 units were classified as "unknown" because the PbN stimulating electrode was found to be outside the anatomically defined taste area in the pons. 3. Electrophysiological responses to sapid solutions of the NaCl (0.1 M), HCl (0.01 M), quinineHCl (0.01 M), sucrose (0.5 M), and Na-saccharin (0.004 M) were then recorded before and after recovery from infusions of procaineHCl into the GN. Both the ipsilateral and contralateral sides of the GN, in that order, received procaine infusions separated by a recovery period of at least 45 min. 4. Analysis of across-unit patterns of response was accomplished with the use of a vector space analysis. With this approach, the response of a given neuron to a given tastant is considered as a coordinate in n-dimensional space, where n is the number of neurons tested. The responses to each stimulus generate vectors whose length relates to the overall magnitude of response across the sample and whose relative directionality indicates similarity to other across-unit patterns. Measures derived from this type of analysis were used as input in a multidimensional scaling (MDS) analysis designed to summarize the organization of the across-unit patterns of response generated by the taste stimuli. This type of analysis creates a "taste space" in which similar across-unit patterns of response are placed close together and dissimilar patterns are placed far apart.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 7472330 TI - Site of interaction between saccade signals and vestibular signals induced by head rotation in the alert cat: functional properties and afferent organization of burster-driving neurons. AB - 1. Extracellular spikes of burster-driving neurons (BDNs) were recorded within and immediately below the prepositus hypoglossi nucleus in the alert cat. BDNs were characterized by short-latency activation after stimulation of the contralateral vestibular nerve (latency: 1.4-2.7 ms) and the ipsilateral superior colliculus (latency: 1.7-3.5 ms). Convergence of vestibular and collicular inputs was found in all of 85 BDNs tested. Firing of BDNs increased during contralateral horizontal head rotation and decreased during ipsilateral rotation. A burst of spikes was induced in association with contralateral saccades and quick phases of nystagmus. 2. BDNs showed irregular tonic discharges during fixation. There was no significant correlation between the firing rate during fixation and horizontal or vertical eye position in most BDNs. During horizontal sinusoidal head rotation, the change in firing rate was approximately proportional to and in phase with contralateral head velocity. The phase lag of the response relative to head angular velocity was 13.8 +/- 20.1 degrees (mean +/- SD) at 0.5 Hz and 7.2 +/- 13.5 degrees at 0.2 Hz on the average. The gain was 0.88 +/- 0.25 (spikes/s)/(degrees/s) at 0.5 Hz and 1.19 +/- 0.49 (spikes/s)/(degrees/s) at 0.2 Hz. 3. Quantitative analysis of burst activity associated with saccades or quick phases indicated that the ON direction of BDNs was contralateral horizontal. The number of spikes in the burst was linearly related to the amplitude of the contralateral component of rapid eye movements. The slope of regression line was, on the average, 1.14 +/- 0.48 spikes/deg. There was no significant difference between the mean slopes for saccades and quick phases. The number of spikes depended on the difference between initial and final horizontal eye positions and not on the absolute eye position in the orbit. The mean burst firing rate was proportional to the mean velocity of the contralateral component of rapid eye movements. The slope of the regression line was 0.82 +/- 0.34 (spikes/s)/(degrees/s). Significant correlation was also found between intraburst instantaneous firing rate and instantaneous component eye velocity. 4. Objects presented in the contralateral visual field elicited a brief burst of spikes in BDNs independent of any eye movement. Contralateral saccades to the target were preceded by an early response to the visual stimulus and subsequent response associated with eye movement. 5. Excitation of BDNs produced by stimulation of the ipsilateral superior colliculus was facilitated by contralateral horizontal head rotation. Therefore saccadic signals from the superior colliculus to BDNs may be augmented by vestibular signals during head rotation.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 7472331 TI - Cholinergic modulation of cortical oscillatory dynamics. AB - 1. The effect of cholinergic modulation on cortical oscillatory dynamics was studied in a computational model of the piriform (olfactory) cortex. The model included the cholinergic suppression of neuronal adaptation, the cholinergic suppression of intrinsic fiber synaptic transmission, the cholinergic enhancement of interneuron activity, and the cholinergic suppression of inhibitory synaptic transmission. 2. Electroencephalographic (EEG) recordings and field potential recordings from the piriform cortex were modeled with a simplified network in which cortical pyramidal cells were represented by excitatory input/output functions with gain parameters dependent on previous activity. The model incorporated distributed excitatory afferent input and excitatory connections between units. In addition, the model contained two sets of inhibitory units mediating inhibition with different time constants and different reversal potentials. This model can match effectively the patterns of cortical EEG and field potentials, showing oscillatory dynamics in both the gamma (30-80 Hz) and theta (3-10 Hz) frequency range. 3. Cholinergic suppression of neuronal adaptation was modeled by reducing the change in gain associated with previous activity. This caused an increased number of oscillations within the network in response to shock stimulation of the lateral olfactory tract, effectively replicating the effect of carbachol on the field potential response in physiological experiments. 4. Cholinergic suppression of intrinsic excitatory synaptic transmission decreased the prominence of gamma oscillations within the network, allowing theta oscillations to predominate. Coupled with the cholinergic suppression of neuronal adaptation, this caused the network to shift from a nonoscillatory state into an oscillatory state of predominant theta oscillations. This replicates the longer term effect of carbachol in experimental preparations on the EEG potential recorded from the cortex in vivo and from brain-slice preparations of the hippocampus in vitro. Analysis of the model suggests that these oscillations depend upon the time constant of neuronal adaptation rather than the time constant of inhibition or the activity of bursting neurons. 5. Cholinergic modulation may be involved in switching the dynamics of this cortical region between those appropriate for learning and those appropriate for recall. During recall, the spread of activity along intrinsic excitatory connections allows associative memory function, whereas neuronal adaptation prevents the spread of activity between different patterns. During learning, the recall of previously stored patterns is prevented by suppression of intrinsic excitatory connections, whereas the response to the new patterns is enhanced by suppression of neuronal adaptation. PMID- 7472332 TI - Development of voltage-activated potassium currents in cultured cerebellar granule neurons under different growth conditions. AB - 1. The functional expression of two potassium currents in cultured cerebellar granule cells was investigated with the whole cell patch-clamp technique in relation to development and growth condition. Cells were grown in medium containing different concentrations of potassium: 25 mM (K25) and 40 mM (K40), together referred to as "high K+"; 10 mM (K10) or "low K+"; and K10 with 100 microM N-methyl-D-aspartate (KNMDA). All conditions are known to influence maturation and survival of granule cells in culture. 2. At 2 days in vitro (DIV) the membrane capacitance, taken as index of membrane surface area, was the same for cells grown in each growth condition. At 7-9 DIV it had increased in each condition, but to a substantially larger extent in cells grown in KNMDA, K25, and K40 than in cells grown in K10. During development the input resistance only decreased in cells grown in KNMDA and high K+. 3. A delayed potassium current (IK) and a fast transient potassium current (IA) could both be recorded at 2 DIV in each growth condition, although a few neurons only expressed the IK. The IK was partially suppressed by tetraethylammonium (5 mM), whereas IA was predominantly sensitive to 4-aminopyridine (5 mM). 4. Normalized for cell capacitance, the specific IA conductance hardly changed during development in cells grown in high K+ and KNMDA. Cells in K10, however, displayed an IA with totally different properties in 23 of 24 cells; the specific IA conductance in these cells was considerably smaller at 7-9 DIV, suggesting a deletion of these channels during development.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7472333 TI - Motor neuronal control of tail-directed and head-directed siphon responses in Aplysia californica. AB - 1. Cutaneous stimulation of opposite ends of the body causes qualitatively different siphon responses: tail stimulation causes flaring and backward bending (the siphon T response), whereas head stimulation causes constriction and slight anterior bending (the siphon H response). This paper characterizes the motor neuronal control of siphon T and siphon H responses. 2. The siphon response to tail nerve (p9) shock in a semi-intact preparation was indistinguishable from the siphon T response in intact or parapodectomized animals. Similarly, the siphon response to head nerve (c2) shock in this preparation was indistinguishable from the siphon H response in intact or parapodectomized animals. 3. Central siphon motor neurons (SMNs) were found to cause a wider variety of movements than previously reported. The movements produced by the LFSB cells strongly resemble the flaring response of the siphon to tail or tail nerve stimulation. The movements produced by RDS and LDS1 resemble components of the constricting response of the siphon to head or head nerve stimulation. 4. Among central SMNs, the LFSB cells show the strongest activation by posterior stimulation, whereas RDS and LDS1 show the strongest activation by anterior stimulation. The LFSA cells, which produce much weaker siphon constriction, are only activated slightly by posterior stimulation and are inhibited by anterior stimulation. Peripheral SMNs are inhibited by stimulation of head and tail nerves, and thus their activity does not directly contribute to siphon T and H responses. 5. Artificially activating central SMNs with the pattern of activity previously exhibited after tail or head nerve stimulation indicated the sufficiency of the LFSB cells for the siphon T response, and of RDS and LDS1 for the siphon H response. 6. Dramatic behavioral deficits produced by hyperpolarizing the LFSB cells during tail nerve stimulation, or by hyperpolarizing RDS and LDS1 during head nerve stimulation, indicated the necessity of these cells for the expression of directed siphon responses to tail or head stimulation, respectively. 7. Because of their apparent necessity and sufficiency for directional siphon responses to anterior and posterior stimulation, these few cells provide well defined vantage points for studying neural mechanisms underlying the motor control and transformation of siphon responses. The four LFSB cells offer a special advantage for cellular analysis because they form a homogeneous functional unit in which any sampled LFSB cell can be used as a precise monitor of the total motor output underlying the siphon T response.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 7472334 TI - Discharge patterns of hippocampal theta-related cells in the caudal diencephalon of the urethan-anesthetized rat. AB - 1. Single-unit discharge patterns of cells in specific nuclei of the caudal diencephalon were characterized in relation to simultaneously recorded field activity from the stratum moleculare of the dentate gyrus according to the criteria that have been used previously to classify cells in the hippocampal formation (including entorhinal cortex), medial septum, and cingulate cortex. Theta (theta)-related cells were classified as 1) tonic theta-ON, if they discharged nonrhythmically and increased their discharge rates during hippocampal theta relative to large, irregular hippocampal field activity (LIA); 2) tonic theta-OFF, if they discharged nonrhythmically and decreased their discharge rates during theta relative to LIA; or 3) phasic theta-ON, if they discharged rhythmically and in phase with ongoing theta, but nonrhythmically during LIA. Cells not meeting any of the above criteria were classified as nonrelated. 2. Recordings were obtained in a total of 127 cells from the caudal diencephalon. Recordings were made in 54 cells from the posterior hypothalamic nucleus (PH), 16 from the supramammillary nucleus (SuM), 20 from the PH/SuM border, and 23 from the medial mammillary nucleus (MM). Recordings were also made from nine cells from the central medial nucleus of the thalamus (CM) and five from the dorsomedial hypothalamic nucleus (DMH). 3. Of the 54 PH cells, 43 (80%) were classified as tonic theta-ON and 11 (20%) as nonrelated. Tonic theta-ON cells in the PH discharged at significantly higher rates during theta, either occurring spontaneously (9.6 +/- 1.7 Hz, mean +/- SE) or elicited with a tail pinch (TP theta; 10.6 +/- 1.9 Hz), than during LIA (3.6 +/- 1.4 Hz). Of the nine CM cells, seven (78%) were tonic theta-ON and two (22%) were nonrelated. Tonic theta-ON cells discharged at significantly higher rates during theta (17.5 +/- 7.8 Hz) or TP theta (18.0 +/- 7.1 Hz) than during LIA (7.3 +/- 4.8 Hz). All DMH cells were nonrelated. 4. Of the 20 PH/SuM border cells, 15 (75%) were classified as tonic theta-OFF and discharged at significantly higher rates during LIA (5.3 +/- 1.5 Hz) than during theta (0.8 +/- 0.4 Hz) or TP theta (0.4 +/- 0.3 Hz). Five (25%) cells in the PH/SuM border were nonrelated. 5. All of the 16 cells (100%) recorded from the body of the SuM were phasic theta-ON. The discharge rates of these cells did not change significantly across hippocampal field states (LIA = 8.3 +/- 1.6; theta = 7.3 +/- 1.6; TP theta = 8.6 +/- 1.7 Hz).(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 7472336 TI - Action of locust neuromodulatory neurons is coupled to specific motor patterns. AB - 1. Many muscles of the locust are supplied by dorsal unpaired median neurons (DUM neurons) that release octopamine and alter the contractions caused by spikes in motor neurons. To determine when these neuromodulatory neurons are normally activated during behaviour, intracellular recordings were made simultaneously from them and from identified motor neurons during the specific motor pattern that underlies kicking. A kick consists of a rapid and powerful extension of the tibia of one or both hind legs that is produced by a defined motor pattern. Only 3 identified DUM neurons of the 20 in the metathoracic ganglion spike during a kick, and they supply muscles involved in generating the kick. Their spikes occur in a distinctive and repeatable pattern that is closely linked to the pattern of spikes in the flexor and extensor tibiae motor neurons. When the extensor and flexor muscles cocontract, these three DUM neurons produce a burst of spikes at frequencies that can rise to 25 Hz, and with the number of spikes (3-15) related to the duration of this phase of the motor pattern. The spikes stop when the flexor muscle is inhibited and therefore before the tibia is extended rapidly. The other DUM neurons which supply muscles that are not directly involved in kicking are either inhibited or spike only sporadically. 2. The activation of a specific subset of DUM neurons during kicking may thus be timed to influence the action of the muscles that participate in this movement and appear to be controlled by the same circuits that determine the actions of the participating motor neurons. These modulatory neurons thus have specific individual actions in the control of movement. PMID- 7472335 TI - NMDA-induced burst discharge in guinea pig trigeminal motoneurons in vitro. AB - 1. The responses of guinea pig trigeminal motoneurons (TMNs) to N-methyl-D,L aspartate (NMA) were studied using brain stem slice preparations and whole cell patch-clamp (n = 89) or conventional microelectrode (n = 22) recording techniques. The primary goals of this study were to determine whether N-methyl-D aspartate (NMDA) receptor activation would produce spontaneous bursting activity in TMNs and, if so, the underlying mechanisms responsible for the generation of these bursts. 2. Bath-applied NMA (100-300 microM, n = 80) in standard perfusion medium elicited depolarization, increase in apparent input resistance (Rinp), and rhythmic burst discharges (1-90 s in duration) from TMNs. These effects were blocked by the NMDA receptor antagonist DL-2-amino-5-phosphonopentanoic acid (AP5, 30 microM, n = 6), but not by the non-NMDA receptor antagonist 6-cyano-7 nitroquinoxaline-2,3-dione (CNQX, 5-10 microM, n = 10). Furthermore, the burst inducing effect of NMA was not mimicked by the non-NMDA receptor agonists kainate (KA, 5-10 microM, n = 6) and (+/-)-alpha-amino-3-hydroxy-5-methylisoxazole-4 propionic acid (AMPA, 5-10 microM, n = 5). 3. In tetrodotoxin (TTX) treatment conditions (n = 13), NMA elicited depolarization, an increase in apparent Rinp, and rhythmic membrane potential oscillations without action potential bursts (i.e., plateau potentials), suggesting that the effects of NMA observed in the TTX-free condition resulted from activation of postsynaptic NMDA receptors. 4. Graded depolarization of neurons (n = 20) by intracellular direct current injection generally led to a graded increase in frequency and duration of the NMA induced bursts and plateau potentials until these rhythmic events eventually became transformed into continuous spike discharge and maintained depolarization, respectively. Removal of Mg2+ from the perfusion medium (n = 11) also turned the bursts and plateau potentials into continuous spike discharge and maintained depolarization, respectively. 5. The effects of NMA on the current-voltage (I-V) curve after a depolarizing ramp voltage-clamp command (15-20 mV/s) were examined (n = 40). Under NMA (100-300 microM) conditions, the I-V relationship exhibited a region of negative slope conductance (NSC) between -60 and -35 mV, thus making the I-V relationship N-shaped. The NSC was abolished by AP5 (30 microM, n = 8), but not by CNQX (5-10 microM, n = 6). The I-V relationship in AMPA (3-10 microM, n = 5) or KA (3-10 microM, n = 5) was almost linear between -80 and -30 mV.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 7472337 TI - Phasic cutaneous input facilitates locomotor recovery after incomplete spinal injury in the chick. AB - 1. Walking and swimming of hatchling chicks was videotaped before hemisection of the left thoracic cord and thereafter at regular intervals, for up to 2 wk. With the use of kinematic techniques, movements of the left knee and ankle were quantified to assess recovery of the ipsilateral leg during walking and swimming trials. To study the effects of exteroceptive (cutaneous) feedback in the absence of limb loading, one group of animals was also provided with cutaneous stimulation during swimming in the form of neutrally buoyant tubes that only contacted the foot during the retraction (extension) phase of the swim cycle. 2. One day after hemisection, for both swimming and walking, the left knee failed to extend normally, and the ankle joint remained hyperextended. During walking, all chicks adopted an asymmetric gait, whereas during swimming the left leg remained retracted and motionless. 3. Over the next 2 wk, knee extension and ankle flexion during walking recovered to normal preoperative values, but neither measure returned to preoperative values during swimming trials. However, when chicks were provided with phasic cutaneous stimulation during swimming trials, they showed improvements in leg motion as soon as 5 days after hemisection. Temporary removal of the cutaneous stimulation during swimming (5 days after hemisection) resulted in reduced limb action. However, 14 days after hemisection, the improvement in limb motion was retained even when the cutaneous stimulation was not provided. 4. Improvement in leg motion after swim training with phasic cutaneous stimulation took the form of increased extension of the limb during retraction. Possible neurophysiological mechanisms for this behavior include reflex reinforcement of limb extensor activity in response to cutaneous stimulation of the foot. Repeated exposure to phasic stimulation during swimming trials results in a permanent alteration in limb action. Thus increased cutaneous afferent inputs, even in the absence of limb loading, can facilitate locomotor recovery after spinal cord injury. PMID- 7472339 TI - Prolonged GABA responses in dentate granule cells in slices isolated from patients with temporal lobe sclerosis. AB - 1. Medial temporal lobe sclerosis is a common pathological finding in patients with medically intractable temporal lobe epilepsy. This disease is characterized by extensive cell loss in the hilus and the hippocampal CA1 and CA3 cell fields in addition to synaptic reorganization throughout the dentate gyrus. 2. The dentate granule cells from hippocampal slices of patients diagnosed with medial temporal lobe sclerosis exhibit reduced synaptic inhibition with concommitant hyperexcitability. These physiological changes were studied relative to the hippocampi of patients with temporal lobe tumors in which the cell loss and synaptic reorganization are not seen. 3. We attempted to determine if this disinhibition was because of changes in the postsynaptic sensitivity to the inhibitory neurotransmitter gamma-aminobutyric acid (GABA) by studying the responses to exogenously applied transmitter. As in rodents, the GABA responses in human dentate granule cells studied at the resting membrane potential were depolarizing and were mediated primarily by GABAA receptors. In many cases, these depolarizing GABA responses could trigger action potentials. Thus in some situations, GABA could act as an excitatory neurotransmitter. 4. We found that GABAA receptor-mediated responses in the sclerotic hippocampi were approximately 80% longer than in the comparison population. This difference was not because of changes in either the GABA reversal potential or the GABA-induced conductance change. The data support the hypothesis that the GABA transport system is impaired in sclerotic tissue: application of the GABA uptake inhibitor NNC711 (a tiagibine derivative) greatly prolonged the GABA responses in the tumor-related temporal lobe epilepsy tissue, but had little effect on the sclerotic tissue. PMID- 7472338 TI - Voltage-gated calcium channels in CNS white matter: role in anoxic injury. AB - 1. The effect of Ca2+ channel antagonists on the extent of anoxia-induced white matter injury was studied in the rat optic nerve, a white matter tract. Compound action potentials (CAPs) were recorded before and after a standard 60-min anoxic period to assess the extent of anoxic injury. 2. The L-type Ca2+ channel antagonists verapamil (90 microM), diltiazem (50 microM), and nifedepine (2.5 microM) significantly protected the rat optic nerve from anoxic injury. Mean recovery of CAP area was 51.3 +/- 3.0% (mean +/- SE, n = 8, P < 0.01), 65.6 +/- 5.3% (n = 8, P < 0.01), and 54.3 +/- 6.1% (n = 8, P < 0.01), respectively. Mean CAP recovery under control conditions was 35.2 +/- 0.3 (n = 33). 3. Simultaneous block of L-type and N-type Ca2+ channels by coapplication of 50 microM diltiazem and 1 microM SNX-124 [synthetic omega-conotoxin (CgTx) GVIA], resulted in postanoxic CAP recovery of 73.6 +/- 6.0% (n = 12), significantly larger than CAP recovery in diltiazem alone (P < 0.001). Block of CgTx MVIIC-sensitive channels in addition to L-type and N-type channels by coapplication of 50 microM diltiazem + 1 microM SNX-230 + 1 microM SNX-124 failed to produce any additional increase in CAP recovery (71.3 +/- 5.6%, n = 8). Application of 1 microM SNX-124 alone did not significantly protect against anoxic injury (CAP recovery, 36.3 +/- 2.9%, n = 10).(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7472340 TI - Epileptiform activity in the dentate gyrus during low-calcium perfusion and exposure to transient electric fields. AB - 1. The dentate gyrus fails to develop epileptiform activity in many experimental models of epilepsy, including the in vitro low-Ca2+ model. Although manipulating the K+ concentration or osmolality of normal low-Ca2+ perfusion mediums can enhance the propensity of the dentate gyrus to develop seizure activity, the specific mechanisms contributing to this change are still under investigation. Identification of these mechanisms should improve our understanding of epileptogenesis and of the factors contributing to the propensity for seizure discharge in other tissues. 2. In the present experiments we used externally generated electric fields to depolarize the somata of large populations of dentate granule cells during exposure to a perfusion medium with no added Ca2+ (low-Ca2+ medium). Uniform electric fields were generated across an individual slice by passing current between two parallel AgCl-coated silver wires placed on the surface of the artificial cerebral spinal fluid. The wires were positioned to straddle the slice such that the current flow was parallel to the dendrosomatic axis of the cell population under investigation. 3. Under control conditions (low Ca2+ medium, no applied field), stimulation of the dentate hilus evoked a single antidromic population spike in 89% of the slices studied (n = 27). During application of electric fields the same stimulus evoked epileptiform activity in all trials. Well-formed bursts first occurred at an average field intensity of +22.9 +/- 2.5 (SE) mV/mm (n = 24). The amplitude of individual spikes and the total number of spikes, within a burst increased in a graded fashion as the magnitude of the applied field was increased. 4. High field intensities evoked epileptiform activity in the absence of a synchronizing antidromic stimulus. These field-induced bursts occurred after a progressive buildup of rhythmic activity recorded in the extrasomatic space and could persist for the entire duration of an applied field, lasting for several seconds. The average field intensity required to produce a threshold burst was +84.6 +/- 3.6 mV/mm (n = 24). 5. In 11% of trials (3 of 27) the dentate gyrus exhibited poorly developed antidromic bursting without the application of depolarizing electric fields. These bursts were completely suppressed by hyperpolarizing fields in the range of -10 to -20 mV/mm. 6. The results of this investigation support the hypothesis that granule cell sensitivity to nonsynaptic interactions is adequate to support bursting in a normal low-Ca2+ medium, but bursting fails to occur because these cells are normally too hyperpolarized relative to their action potential threshold.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 7472341 TI - Cerebellar and cerebral inputs to corticocortical and corticofugal neurons in areas 5 and 7 in the cat. AB - 1. In the parietal cortex (Px, areas 5 and 7), the organization and characteristics of cerebellar and cerebral inputs and their effects on efferent neurons were investigated with the use of intracellular and extracellular recording techniques in the anesthetized cat. 2. Evoked field potential analysis revealed that two regions of the Px, the caudal bank of the ansate sulcus (Ans. S.) and the crown of the suprasylvian gyrus (Ssyl. G.), received converging input from the dentate and the interpositus nucleus. The cerebellar input to the caudal bank of the Ans. S. was relayed via the ventrolateral region of the ventroanterior-ventrolateral (VA-VL) complex of the thalamus, whereas the cerebellar input to the crown of the Ssyl. G. was relayed via the dorsomedial region of the VA-VL complex. 3. A total of 176 neurons was recorded intracellularly in the Px to examine inputs from the cerebellum. Of these, 72 neurons were corticocortical neurons projecting to the motor cortex (Mx), and 48 were corticofugal neurons to the pontine nucleus (PN). Intracellular staining with horseradish peroxidase revealed that the former corticocortical neurons were layer III pyramidal neurons and the latter corticofugal neurons were layer V pyramidal neurons. 4. Stimulation of the brachium conjunctivum (BC) produced di- or polysynaptic excitatory postsynaptic potentials (EPSPs) in corticocortical neurons projecting to the Mx and corticofugal neurons to the pontine nucleus in the Px. The characteristics of BC-evoked EPSPs were different between the bank of the Ans. S. and the crown of the Ssyl. G. In the bank of the Ans. S., the slope of the rising phase of the BC-evoked EPSPs was steeper, and their minimum latency was shorter by 0.8 ms than those in the crown of the Ssyl. G. These differences may reflect differences in the terminal distribution and conduction velocity of the thalamocortical fibers relaying cerebellar input to these two parietal areas. 5. Stimulation of the Mx produced mono- or disynaptic EPSPs in both corticocortical neurons projecting to the Mx and corticofugal neurons projecting to the pontine nucleus in the Px. For each neuron, effective sites for inducing EPSPs were distributed very widely and sometimes covered both areas 4 and 6. Extensive corticocortical projection from the Mx to the Px was confirmed by injection of an anterograde tracer into the Mx. 6. These data indicate that neurons in the Px receive inputs from both the cerebellum and the Mx and send outputs to the Mx and the cerebellum.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 7472343 TI - Vertical vestibular input to and projections from the caudal parts of the vestibular nuclei of the decerebrate cat. AB - 1. To investigate the type of vestibular signals that neurons in the caudal parts of the vestibular nuclei transmit to the cerebellum and spinal cord, we studied their responses to natural vestibular stimulation in vertical planes in decerebrate cats with the caudal cerebellum removed. Most neurons were in the caudal half of the descending vestibular nucleus, the remainder at corresponding levels of the medial nucleus or the medial-descending border. 2. Dynamics of the responses of spontaneously firing neurons were studied with sinusoidal tilts delivered at 0.05-1 Hz near the plane of body rotation that produced maximal modulation of the neuron's activity (response vector orientation). For most neurons the predominant vestibular input could be identified as coming from otolith organs (46%) or vertical semicircular canals (37%). Some neurons had otolith+canal convergence (9%) and others either had such converging input or some other form of central processing (8%). 3. Gain and phase of the responses of otolith neurons were comparable with values obtained in earlier studies on Deiters' nucleus and the rostral descending nucleus. Many canal neurons had a steeper gain slope and more advanced phase than observed previously for more rostral neurons. This may be due to more irregular afferent input to many neurons or to the absence of the vestibulocerebellum. 4. Response vector orientations of canal neurons were closely bunched near the planes of the ipsilateral vertical canals. The small number of contralaterally projecting vectors showed evidence of convergence between the two contralateral vertical canals. As is the case elsewhere in the vestibular nuclei, there was no evidence of convergence from bilateral vertical canals. Response vector orientations of otolith neurons were restricted to the roll quadrants; the majority pointed ipsilaterally. 5. Antidromic stimulation with an electrode in the restiform body or with several electrodes in the dorsal half of the white matter of the upper cervical cord was used to identify neurons projecting to the cerebellum and spinal cord, respectively. A substantial number of spontaneously firing neurons projected to the cerebellum, but there were few spontaneously active vestibulospinal neurons. The properties of the vestibular input to cerebellar-projecting neurons were the same as those of the population as a whole, but the effect of tilt on vestibulospinal neurons appeared weak or absent. 6. Many neurons were inhibited by stimulation of the restiform body. We suggest that this is mainly due to stimulation of the axons of vestibulocerebellar Purkinje cells. 7. Our results demonstrate a robust vertical vestibular input to the caudal parts of the vestibular nuclei.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 7472342 TI - Human cortical regions activated by wide-field visual motion: an H2(15)O PET study. AB - 1. Several areas in the monkey dorsal visual pathway, including the dorsal part of the medial superior temporal area, have been found to contain cells responding to movements of a wide visual field and are suggested to be involved in analyzing self-induced motion information. In the present study, positron emission tomography was used to localize human cortical regions responding to wide-field visual motion. Changes in regional cerebral blood flow (rCBF) were measured when subjects maintained fixation and viewed low-contrast (0.15 log units brighter than the background) dots subtending 80 x 80 degrees and moving either coherently or incoherently. Brain foci were localized after activity in a fixation-only paradigm was subtracted from that in the two moving dot paradigms. 2. Both the coherent and incoherent movements significantly activated the primary/secondary visual cortex and surrounding visual areas in the cuneus and superior occipital gyrus. Subtraction of images between the coherent and incoherent movements showed that the activity caused by the two types of movement was comparable in these early visual cortical regions. 3. In the lateral occipitotemporoparietal cortex, the coherent movement specifically activated two separate areas; a posterior focus was located at the border of the right occipitotemporal gyri, and a dorsoanterior focus was located bilaterally in the temporoparietal cortex. The incoherent movement did not activate these regions. 4. A fine anatomic localization using individual magnetic resonance images was performed for the bilateral activation in the temporoparietal cortex, which was found to be located mainly in the depth of the inferior parietal lobule and a small portion of the superior and middle temporal gyri. 5. Both the coherent and incoherent movements activated a part of the superior parietal lobule located within the intraparietal sulcus (Brodmann area 7). The bilateral foci activated by the coherent movement were located more anteriorly than the focus activated by the incoherent movement. Subtraction images between the coherent and incoherent movements, however, did not reveal any significant rCBF increases in the superior parietal lobule. 6. Several other cortical regions known to be involved in visuospatial and visuomotor functions were also activated by the coherent movement, including the frontal eye field (Brodmann area 8) and premotor cortex (Brodmann area 6) in the frontal lobe. 7. The posteriorly located activation at the border of occipito temporal gyri corresponds to the homologue of the middle temporal area reported in previous activation studies using small to medium-sized motion stimuli. The bilateral activation in the inferior parietal lobule appeared to rely on wide field motion stimulation. PMID- 7472344 TI - Calcium channel involvement in GABAB receptor-mediated inhibition of GABA release in area CA1 of the rat hippocampus. AB - 1. Experiments were performed in rat hippocampal slices to examine the nature of GABAergic inhibition of inhibitory synaptic transmission. In these experiments the effects of the gamma-aminobutyric acid-B (GABAB) receptor agonist, baclofen, and of subtype-selective calcium channel blockers were tested with the use of intracellular recordings of evoked inhibitory postsynaptic potentials (IPSPs) and whole cell recordings of spontaneous GABAergic inhibitory postsynaptic currents (IPSCs). 2. Baclofen inhibited evoked and spontaneous (action-potential dependent) monosynaptic GABAA-mediated IPSPs and IPSCs but had no effect on the frequency of tetrodotoxin-resistant (action-potential-independent) miniature IPSCs recorded in CA1 pyramidal neurons. 3. Depolarizing GABAergic synaptic terminals by raising the extracellular potassium concentration caused an increase in action-potential-independent miniature IPSC frequency that could be inhibited by either baclofen or cadmium, a blocker of voltage-dependent calcium channels. In addition, under these depolarizing conditions, cadmium occluded the baclofen inhibition of miniature IPSCs. These data suggest that baclofen reduces only depolarization-induced, not quantal, GABA release and that it does so by decreasing presynaptic voltage-dependent calcium influx. 4. Experiments with subtype-selective calcium channel blockers demonstrate that the presynaptic action of baclofen was mediated through both omega-conotoxin-GVIA-sensitive and omega-agatoxin-IVA-sensitive, but not dihydropyridine-sensitive calcium channels. PMID- 7472345 TI - Distributed amine modulation of graded chemical transmission in the pyloric network of the lobster stomatogastric ganglion. AB - 1. In the pyloric network of the lobster stomatogastric ganglion, graded synapses organize the network output. The amines dopamine (DA), serotonin, and octopamine each elicit a distinctive motor pattern from a quiescent pyloric network. We have examined the effects of these amines on the graded synaptic strengths between the six major types of neurons of this network to understand how amine modulation of synaptic strength contributes to the amine-induced motor patterns. Here we tested amine affects at 10 different graded chemical synapses of the pyloric network. We show that each amine has a statistically different spectrum of distributed effects across the network synapses. 2. Under our control conditions (isolated pairs of neurons, removal of modulatory input), most of the graded chemical synapses were weak and some synapses were nonfunctional. The output synapses of the ventricular dilator (VD) neuron were significantly stronger than the other synapses. 3. DA altered the synaptic strength of every graded chemical synapse. This amine strengthened the weak chemical output synapses of the anterior burster (AB), lateral pyloric (LP), and pyloric constrictor (PY) neurons and weakened (and in some cases abolished) the strong chemical output synapses of the VD neuron. The AB-->inferior cardiac neuron (IC) and PY-->IC graded chemical synapses were nonfunctional under our control conditions; DA activated these silent synapses. 4. Serotonin enhanced the AB's output chemical synapses but weakened all the other graded chemical synapses examined. Octopamine's effects were much weaker than those of the other two amines. It enhanced the AB-->LP synapse and the LP's output synapses and weakly strengthened the AB-->PY, VD- >LP, and VD-->PY synapses. 5. The amines alter the input resistance of many of the pyloric neurons, and this could contribute to the observed changes in synaptic strength by altering passive current flow between input and output sites in the cells. However, the input resistance changes were relatively small compared with the changes in synaptic strength and cannot alone account for the synaptic modulation. In some cases the sign of the input resistance change was inconsistent with the change in synaptic strength. Thus the amines appear to modify synaptic transmission directly in this system. 6. This study completes our description of amine effects on all the graded synapses of the pyloric network. We summarize our present and earlier work to show that modulators can reconfigure the entire synaptic organization of a neural network by acting at many distributed synaptic sites.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 7472346 TI - 5-HT1B/1D agonist CGS-12066B attenuates clasp knife reflex in the cat. AB - 1. The effect of intrathecal injection of the selective serotonin (5-HT)1B/1D receptor agonist CGS-12066B maleate (825 nmol) was assessed on stretch-evoked clasp knife inhibition of hindlimb ankle extensor muscle reflex force in precollicular decerebrate cats in which neural transmission in dorsolateral spinal pathways was blocked bilaterally by focal cooling. 2. During cold block, ramp and hold stretches of the medial gastrocnemius muscle (MG) evoked only a brief reflex excitation that was followed by powerful, long-lasting inhibition (the clasp knife reflex). Both the amplitudes of peak force evoked by the ramp and sustained force output during the last 500 ms of the hold phase of the stretch were depressed by > 50%. 3. Reflex force output during the hold portion of stretch was significantly improved on postdrug cold block trials, although peak force remained depressed. CGS-12066B did not significantly alter stretch evoked force output in decerebrate cats when spinal cord neural transmission was unimpaired. 4. These data suggest that selective 5-HT1B/1D agonists may be of therapeutic usefulness in the treatment of reflex disorders arising from partial spinal cord injury. PMID- 7472347 TI - Vectorial coding of movement: vision, proprioception, or both? AB - 1. Subjects were asked to point toward visual targets without visual reafference from the moving hand in two conditions. In both conditions the pointing fingertip was viewed only before movement onset. 2. In one condition, the pointing fingertip was viewed through prisms that created a visual displacement without altering the view of the target. In another experimental condition, vision of the fingertip was not displaced. Comparison of these two conditions showed that virtually shifting finger position before movement through prisms induced a pointing bias in the direction opposite to the shift. The extent of this pointing bias was about one third of the prismatic shift applied to the fingertip. 3. Analysis of movement initial direction demonstrated that it was also less deviated than predicted from the prismatic shift. In addition, the reaction time and movement time of the reaching movement were increased. 4. This result is interpreted in the framework of the vectorial coding of reaching movement. Proprioception and vision provide two possible sources of information about initial hand position, i.e., the origin of the movement vector. The question remains as to how these two sources of information interact in specifying initial hand position when they are simultaneously available. 5. Our results are thus discussed with respect to a visual-to-visual movement vector hypothesis and a proprioceptive-to-visual vector hypothesis. It is argued that the origin of the putative movement vector is encoded by weighted fusion of the visual and the proprioceptive information about hand initial position. PMID- 7472348 TI - Withdrawal from the endogenous steroid progesterone results in GABAA currents insensitive to benzodiazepine modulation in rat CA1 hippocampus. AB - 1. The withdrawal properties of the endogenous steroid progesterone (P) were tested in female rats as a function of benzodiazepine modulation of gamma aminobutyric acid-A (GABAA)-gated current with the use of the whole cell patch clamp technique on acutely dissociated CA1 hippocampal neurons. In a previous study, this steroid was shown to exhibit withdrawal properties, behaviorally. 2. One day withdrawal from in vivo administration of physiological doses of P (5 mg ip, 5 days/wk for 3 withdrawal cycles) or its metabolite, the GABAA modulator 3 alpha-hydroxy-5 alpha-pregnan-20-one (3 alpha,5 alpha-THP or allopregnanolone, 20 mg/kg ip) prevented the normally potentiating effect of lorazepam (LZM; 10(-7) 10(-4) M) on GABAA-gated current. Withdrawal from 500 micrograms P administered concomitantly with 2 micrograms 17 beta-estradiol also markedly diminished LZM potentiation of GABAA current. This effect was seen only after three withdrawal cycles. 3. P withdrawal produced no inhibitory effect on either basal levels of GABAA-evoked current, the GABAA EC50, or barbiturate (+/-Pentobarbital, 10(-7) 10(-4) M) modulation of this parameter. 4. The effect of steroid withdrawal on LZM modulation of GABAA-evoked current was blocked by picrotoxin as well as by indomethacin, a drug that prevents conversion of P to its metabolite, the GABAA modulator 3 alpha,5 alpha-THP. These results suggest that the withdrawal properties of P may be due to changes in GABAA receptor function produced by 3 alpha,5 alpha-THP. PMID- 7472349 TI - Tonic transmitter release in a graded potential synapse. AB - 1. We studied graded synaptic transmission in the fly photoreceptor-interneuron synapse by using intracellular in situ recordings from pre- and postsynaptic cells. 2. A large presynaptic hyperpolarization after light adaptation, caused by the activation of the electrogenic Na+/K+ pump, drastically reduced the conspicuous postsynaptic dark noise. At the same time, the postsynaptic neurons depolarized, with an increase of input resistance of 5-10 M omega. 3. The spectral characteristics of the postsynaptic membrane noise in dark and during noise reduction, together with the other results, suggested that the transmitter release decreased dramatically approximately 12 mV below the resting potential of the presynaptic photoreceptors. 4. During the postsynaptic noise reduction, the saturated and subsaturated first-order visual interneuron responses were increased up to 9 mV with a time constant of recovery of approximately 10 s. This increase was shown to be caused by the negative shift of the reversal potential of the transmitter-gated (mainly Cl-) conductance, caused apparently by the reduced transmitter input. 5. The results strongly suggest that the photoreceptor transmitter release in fly is tonic, even in dark, and further support the modulation of the synaptic voltage transfer by postsynaptic Cl- extrusion. PMID- 7472350 TI - Preferential correlations of a medullary neuron's activity to different sympathetic outflows as revealed by partial coherence analysis. AB - 1. In vagotomized, paralyzed, decerebrate cats, simultaneous recordings were taken from one or more sympathetic nerves [cervical sympathetic (CS), inferior cardiac (IC), splanchnic (SP)] and from medullary neurons in vasomotor-related regions. Coherence analyses were used to ascertain the presence of sympathetic rhythms (2-6 Hz or "3-Hz rhythm," 7-13 Hz or "10-Hz rhythm") that were correlated between different signals. The occurrence of a significant peak at such a frequency in a unit-nerve coherence spectrum allowed the identification of a medullary neuron as sympathetic related. 2. A serendipitous example is given of a rostral ventrolateral medullary neuron that had significant unit-nerve 10-Hz coherence peaks for three sympathetic nerves (CS, IC, SP); but, as revealed by partial coherence analysis, the unit activity's correlation with one nerve's activity could be partially or completely dependent on its correlation with other nerve activities. Thus in this case the unit-CS and unit-IC coherences at 10 Hz were completely dependent on the SP rhythm, whereas the unit-SP coherence was not significantly affected by the CS and IC rhythms. This asymmetry suggests that the neuron was preferentially connected to SP-generating medullary circuits. 3. This example indicates the strength of partial coherence analysis as a means of studying differential connectivity between medullary sympathetic-related neurons and sympathetic output neuron populations. PMID- 7472352 TI - Fast imaging of [Ca]i reveals presence of voltage-gated calcium channels in dendritic spines of cultured hippocampal neurons. AB - 1. Cultured hippocampal neurons were recorded with a patch pipette containing 100 microM of the calcium indicator Fluo-3, and one of their dendrites, carrying dendritic spines, was visualized with a x100, 1.3-numerical aperture oil objective. Calcium spikes evoked by depolarizing the somata and changes in free dendrite and spine calcium concentrations ([Ca]d and [Ca]s, respectively) were monitored with a cooled charge-coupled device (CCD) camera, acquiring images at a rate of 17-20 ms per frame. In the majority of spine-dendrite pairs, [Ca]s rose faster and to a higher level than the adjacent [Ca]d. Likewise, topical application of glutamate evoked a faster and larger change in [Ca]s than in [Ca]d. The rise of intracellular calcium concentration in response to a depolarizing current pulse, but not in response to glutamate, was reduced in the presence of the calcium antagonist verapamil in both dendrites and spines. It is suggested that dendritic spines possess voltage-gated calcium channels. PMID- 7472351 TI - Odor-induced currents in Xenopus olfactory receptor cells measured with perforated-patch recording. AB - 1. Odor-evoked currents were recorded in Xenopus laevis olfactory receptor neurons (ORNs) by the use of conventional, as well as nystatin and gramicidin perforated, whole cell recording. The odor-evoked current ran down quickly in conventional, but not in perforated, whole cell recording. All three types of recording gave similar values for the amplitude, latency, time-to-peak, recovery time, and reversal potential of the odor-evoked current. 2. A secondary Cl current comprised a significant part of the odor-evoked current (55-65%). ECl measured by gramicidin perforation, which does not alter [Cl-]i, was -2.3 +/- 5.0 (SE) mV, indicating that these neurons maintain a high [Cl-]i and that the secondary Cl current plays an excitatory role in olfactory transduction. PMID- 7472353 TI - Cerebellar nitric oxide is necessary for vestibulo-ocular reflex adaptation, a sensorimotor model of learning. AB - 1. Nitric oxide (NO) production in the nervous system has been implicated in cellular mechanisms of learning and memory. Our study investigates an in vivo sensorimotor model of learning. It demonstrates that a localized vestibulocerebellar injection of the NO synthase inhibitor, L-NG-monomethyl arginine (L-NMMA), which specifically blocks NO production, inhibited the acquisition of adaptive vestibulo-ocular reflex (VOR) gain increases but not gain decreases in the goldfish. 2. Restoration of NO production by concomitant administration of L-arginine (the substrate for NO synthase) and L-NMMA suppressed the inhibitory effect of L-NMMA on adaptive gain increases. 3. This effect of L-NMMA was stereospecific because injection of D-NMMA did not suppress adaptive VOR gain increases. 4. Injection of L-NMMA after VOR adaptation had no effect on retention, failing to alter the postadaptive recovery after a VOR gain increase. 5. In conclusion, acquisition of adaptive VOR gain increases are affected by cerebellar NO inhibition. However, because gain decreases are not, they may involve either non-NO cerebellar or extracerebellar mechanisms. In addition, different processes for acquisition and retention of gain increases may be operating, because inhibition of cerebellar NO affects the acquisition but not the retention phase. PMID- 7472354 TI - Nine members of the myomodulin family of peptide cotransmitters at the B16-ARC neuromuscular junction of Aplysia. AB - 1. Neuromodulation by multiple related peptides with different spectra of physiological effects appears an effective way to integrate complex physiological functions. A good opportunity to examine this issue occurs in the accessory radula closer (ARC) neuromuscular circuit of Aplysia, where, extensive previous work has shown, acetylcholine-induced contractions of the muscle are variously modulated by several families of peptide cotransmitters released under appropriate behavioral circumstances from the muscle's own two motor neurons. 2. In this work we focused on the myomodulins (MMs) released from motor neuron B16. Previous work has characterized MMA (PMSMLRLamide) and MMB (GSYRMMRLamide). We now similarly purified from ARC neuromuscular material and sequenced MMC (GWSMLRLamide), MMD (GLSMLRLamide), MME (GLQMLRLamide), and MMF (SLNMLRLamide). Three additional MMs, MMG (TLSMLRLamide), MMH (GLHMLRLamide), and MMI (SLSMLRLamide), are encoded by a known MM gene. B16 probably synthesizes, and coreleases, all nine MMs. Further MMs have been found in other mollusks. All evidence indicates that the MMs are a major, widely distributed family of molluscan neuropeptides important as neuromuscular modulators and probably also central transmitters or modulators. 3. MM effects on motor neuron B16-elicited ARC muscle contractions were best analyzed as the sum of three distinct actions: potentiation, depression of the amplitude of the contractions, and acceleration of their relaxation rate. We compared the effectiveness of all nine MMs in these respects. We correlated this with their effectiveness in enhancing the L-type Ca current and activating a specific K current in voltage-clamped dissociated ARC muscle fibers, effects we previously proposed to underlie, respectively, the potentiation and the depression of contractions. 4. All nine MMs were similarly effective in enhancing the Ca current and, as far as it was possible to determine, potentiating the amplitude as well as accelerating the relaxation rate of the contractions. 5. In contrast, the MMs' ability to activate the K current and depress the contractions varied greatly. MMB and MMC, in particular, were weak, whereas the other seven MMs were considerably more effective in both respects. 6. Altogether, we were able to explain the potentiating and depressing strengths of the MMs by the magnitude of their modulation of the Ca and K currents, providing further support for our hypothesis that the effects on contraction amplitude are mediated by the effects on the two currents. 7. The net effect on contraction amplitude was determined by the balance between the potentiation and depression. Although most MM concentrations had both potentiating and depressing actions, potentiated contractions predominated at low and depressed contractions (but with accelerated relaxation rate) at high concentrations.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 7472355 TI - Activity of neurons in the medial pontomedullary reticular formation during orienting movements in alert head-free cats. AB - 1. Single unit activities of 236 neurons were recorded in the medial pontomedullary reticular formation during visually triggered orienting gaze shifts in 10 alert cats under head-free conditions using movable tungsten-needle electrodes attached to the skull. The activities were analyzed mainly in relation to the head movement that was triggered by presentation of a light-emitting diode (LED) in one of eight directions separated radially by 45 deg after fixation of the center LED. Of these, 120 neurons were recorded in the pontine reticular formation, chiefly in the nucleus reticularis pontis caudalis, and the remaining 116 were in the medullary reticular formation, chiefly in the nucleus reticularis gigantocellularis. Activities of 65 pontine and 65 medullary neurons were modulated in relation to the dynamic phase of orienting movements ("orienting related neurons"). Activities of the remaining neurons were modulated either irregularly or not at all during orienting movement ("irregular or no-response neurons"). Input from the contralateral superior colliculus and cerebral cortex and projections to the spinal cord were also investigated. 2. Among the orienting related neurons, 62 pontine and 55 medullary neurons showed increases in activity preceding the onset of eye and head movement by 0-155 ms ("pretype"). Three pontine and 10 medullary neurons showed increases in activity only after the onset of movement ("posttype"). Of the pretype neurons, 61 pontine and 51 medullary neurons showed directional preference of activity ("directional" neurons). One pontine and four medullary neurons were classified as "omnidirectional" because these neurons increased activity preceding movements in all directions tested, and no directional preference was apparent. 3. In the pretype-directional cells, the average firing frequency during bursts was correlated with amplitude and angular velocity of head movements. Activities of the directional neurons during movements in the eight different directions could be well fitted with cosine functions in the majority of cases. The preferred directions of most pontine neurons and of about half the medullary neurons, as determined by first-degree sinusoidal regression analysis, were distributed around the ipsiversive horizontal axis. However, there were also a considerable number of neurons whose preferred directions were upward, downward, contraversive, or oblique in the medulla. 4. Among the directional cells preferring ipsiversive horizontal movements, 11 pontine neurons showed activity, the onset of which was locked to visual stimuli with latencies of 40-70 ms, in addition to phasic discharges locked to the onset of movement. This "stimulus locked activity" was sometimes modulated depending on the attentional state of the animal.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 7472356 TI - Comparison of evoked potentials and high-frequency (gamma-band) oscillating potentials in rat auditory cortex. AB - 1. Transient and steady-state (40 Hz) evoked potentials, as well as spontaneous and click-evoked gamma-band oscillations, were recorded from 15 lightly anesthetized rats using an 8 x 8 electrode epipial array covering auditory cortex and adjacent areas to determine and compare the spatiotemporal distributions of these four phenomena. 2. The transient evoked response replicated earlier findings in our laboratory, consisting of an initial biphasic sharp wave in area 41, a similar but delayed biphasic sharp wave in area 36, and more widely distributed slow-wave components. Spatiotemporal analysis supported a model of parallel and asynchronous activation of distinct groups of thalamocortical projections underlying the neurogenesis of these temporal components of the middle-latency auditory evoked potential (MAEP) complex. 3. The 40-Hz response to click trains was superimposed on a steady potential shift (SP), both of which were localized within primary auditory cortex. Epipial distributions of the SP were similar to those of the shortest-latency negative peak in area 41 recorded in the same animals, suggesting similar neural generators. The 40-Hz response was more focal and dissimilar from the SP and any other temporal components of the MAEP complex, suggesting that a unique subpopulation of cells underlies its neurogenesis. 4. Spontaneous gamma-band activity, as assessed by power spectrum analysis, was localized to primary and secondary auditory cortex but had a variable distribution between rats that did not conform to the cytoarchitectonic boundaries within subdivisions of this region. Digital movies computed for individual bursts of gamma-activity indicated a high degree of spatiotemporal variability within and between bursts. 5. Single-trial spectral analysis of click responses indicated an inhibition of gamma-band oscillations during most of the MAEP complex, with subsequent enhanced gamma-activity during the 300- to 350-ms slow-wave component that outlasted the MAEP by approximately 500 ms. The epipial distributions of prestimulus and enhanced poststimulus gamma-oscillations were the same. In contrast to the 40-Hz response to click trains, phase-locking of gamma-oscillations by the single click stimulus was not observed. 6. These results suggest that both the MAEP complex and the steady-state 40-Hz response with its associated SP are highly stereotyped in lightly anesthetized rodent cortex. Their spatiotemporal distributions are probably determined in large part by asynchronous activation of parallel thalamocortical projection systems. Our data suggest no direct link between either the MAEP or the steady-state 40-Hz response to spontaneous or evoked gamma-band oscillations in auditory cortex.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 7472357 TI - Evidence that inhibitory mechanisms mask inappropriate somatotopic connections in the spinal cord of normal rat. AB - 1. The responses to stimulation of the sciatic and saphenous nerves have been studied in 65 pairs of spinal dorsal horn neurons simultaneously recorded at the L2 and L5-L6 lumbar segments of the rat's spinal cord. The neurons were recorded in anesthetized and paralyzed animals. 2. Five- or seven-barreled micropipettes were utilized for recording and for the application of drugs with iontophoresis or micropressure techniques. The drugs used were: strychnine, as a selective antagonist at glycine receptors; sodium glutamate and N-methyl-D-aspartate (NMDA), as agonists at excitatory glutamatergic receptors; glycine, as an agonist at the inhibitory glycine receptor; and the local anesthetic lidocaine, as a reversible local conduction blocker both in the periphery and in the spinal cord. 3. All neurons had cutaneous receptive fields in the ipsilateral hindpaw. Neurons responding exclusively to saphenous stimulation in L2 and to sciatic stimulation in L5-L6 were selected for this study. The responses consisted of bursts of > or = 5 spikes, often partially inserted in a field potential, with latencies of 5.0 +/- 1.1 (SD) and 5.2 +/- 1.2 ms, respectively. The thresholds of stimulation and the response latencies were controlled to be stable throughout the experiments. 4. Eighty-five percent (29 of 35) of the neurons tested in L5-L6 exhibited responses to saphenous stimulation during strychnine microejection in the recorded neurons. The neurons became again unresponsive to saphenous stimulation shortly after the end of strychnine ejection. 5. All the neurons tested in L5-L6 (n = 14) showed a significant increase in background activity and remained unresponsive to saphenous stimulation during glutamate microejection on the recorded neurons. 6. All the neurons tested in L5-L6 (n = 17) showed responses to saphenous stimulation after sciatic nerve block with local anesthetic. The responses to saphenous stimulation disappeared after the effect of local anesthetic ceased. 7. All the neurons tested in L5-L6 (n = 6), in rats with a local block of the sciatic nerve, showed a reversible increase in the background activity during NMDA microejection in the spinal saphenous area at L2. 8. In rats with a local block of the sciatic nerve, the responses evoked in L5-L6 neurons (n = 12) by saphenous stimulation were reduced both in field potential amplitude and in number of spikes during microejection of glycine, and were suppressed during microejection of lidocaine in the spinal saphenous area in L2.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 7472358 TI - Diverse current and voltage responses to baclofen in an identified molluscan photoreceptor. AB - 1. gamma-Aminobuturic acid-B (GABAB) receptors play a role in the mediation of slow inhibitory postsynaptic potentials in mammalian as well as some nonmammalian species. In identified photoreceptors from the marine mollusc Hermissenda, recent evidence has suggested that GABA, as well as the GABAB receptor agonist baclofen, might simultaneously modulate multiple conductances on the postsynaptic membrane. Here, using intracellular current-clamp and single-electrode voltage-clamp techniques, we have characterized responses to baclofen in the B photoreceptors of the Hermissenda eye. 2. Microapplication of baclofen (12.5-62.5 microM) to the terminal branches of the B photoreceptors induced a slow, concentration-dependent hyperpolarization (approximately 3-8 mV) that was accompanied by a cessation of spontaneous action potentials and a positive shift in firing threshold. Both the hyperpolarization and the shift in spike threshold in response to baclofen were attenuated largely by the K+ channel blocker tetraethylammonium chloride (TEA; 50 mM). 3. Bath application of baclofen (100 microM) decreased the amplitude, duration, and the afterhyperpolarization (AHP) of evoked action potentials. Although baclofen's effect on spike duration and amplitude persisted in the absence of extracellular Ca2+, the reduction of the AHP by baclofen was eliminated, suggesting that multiple conductances mediated the baclofen-induced modification of the action potential. 4. Using a single-electrode voltage-clamp technique, microapplication of baclofen to the terminal branches of the B photoreceptor produced a slow, net outward current (< 0.5 nA) that reversed near the equilibrium potential for K+ and shifted to more positive potentials when extracellular K+ was increased, in approximate agreement with the Nernst equation for K+. 5. Baclofen induced an increase in amplitude of the nonvoltage dependent leak conductance (IL), and the increase was blocked by TEA. The baclofen-induced increase of IL was accompanied by an increase in amplitude and a negative shift in the voltage dependence of a slow, steeply voltage-dependent K+ current (IK), which displays selective sensitivity to TEA but does not normally contribute to leak conductance. The amplitude and steady-state inactivation of a fast, transient K+ current, as well as the amplitude of an inwardly rectifying K+ current were unaffected by baclofen. 6. Both the rate of activation as well as the amplitude of a voltage-dependent Ca2+ current (ICa) were reduced by baclofen. The reduction of ICa resulted in a concomitant suppression of a Ca(2+)-dependent K+ current (IK-Ca) that was sufficient to account for the reduction of the AHP after evoked action potentials.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 7472360 TI - Depression of synaptic connections between identified motor neurons in the locust. AB - 1. The fast extensor tibiae motor neuron makes direct excitatory central connections with the posterior group of flexor tibiae motor neurons in the locust metathoracic ganglion. The flexor group has a slow, a fast, and an intermediate motor neuron. The motor neurons are involved in the motor program for kicking and jumping, the defensive and escape behaviors of the locust. An antidromic action potential in fast extensor tibiae motor neuron (FETi) results in a monosynaptic, glutamatergic excitatory postsynaptic potential (EPSP) in each of the flexor motor neurons. 2. A train of 10 antidromic spikes in FETi at frequencies of 1-20 Hz resulted in depression of the amplitude of the EPSP in each of the flexor motor neurons. The depression was not significantly different in the different flexor motor neurons. The depression was greater with higher frequency stimulation and was reduced in low calcium saline. 3. After stimulation at 20 Hz, the EPSP amplitude was depressed by approximately 80%. This did not change when the number of stimuli was increased to 20, when stimulation was done in high calcium saline, or when the frequency of stimulation was increased to 50 or 100 Hz. The recovery from depression was greater after 20-Hz stimulation than at lower frequencies, although the recovery was reduced when the number of stimuli was increased, and also in high calcium saline. 4. In normal saline the depression of the EPSP amplitude was associated with a reduction of the presynaptic spike amplitude at frequencies of > or = 5 Hz. In tetraethylammonium (TEA) saline the width of a TEA-broadened spike was also reduced. The reduction in spike amplitude and spike width correlated with the depression of the EPSP. 5. Certain of these results are consistent with a depletion model of synaptic depression, whereas others are not consistent with this model. The depression may be partly due to an initial depletion of transmitter stores, and partly to modulation of the presynaptic action potential that reduces calcium entry, and therefore transmitter release. The significance of the depression on the motor program for kicking and jumping is discussed. PMID- 7472359 TI - Opioid inhibition in locus coeruleus. AB - 1. Inhibition of locus coeruleus (LC) neurons by opioids is mediated by hyperpolarization associated with an increase in potassium conductance. However, opioids caused an outward current even at potentials more negative than the potassium equilibrium potential in the majority of LC neurons recorded in brain slices cut in the horizontal plane. 2. Whole cell and intracellular recording were made from LC neurons in the slice preparation and currents were measured with a switch clamp amplifier. 3. Local application of [Met]5enkephalin (ME) by iontophoresis resulted in two different effects depending on the site of application. Iontophoresis of ME close to the cell caused an outward current that reversed polarity near the potassium equilibrium potential. Whereas application at a distance > 200 microns resulted in an outward current that did not reverse polarity even at strongly negative potentials. 4. When potassium conductances were blocked with internal Cs and/or external Ba, the opioid current was reduced to < 15% of control. 5. Low sodium solutions were used to reduce sodium-dependent conductances. These solutions hyperpolarized LC neurons themselves, decreased outward currents caused by ME, and shifted the reversal potential of the ME induced current to less negative values. 6. Electrotonic coupling of LC neurons in neonatal animals is thought to synchronize spontaneous activity. In this study, synchronous activity in cells from adult animals also was observed, suggesting that electrotonic coupling in the LC persists into adulthood. 7. Carbenoxolone, an agent thought to block electrotonic coupling, had no effect on the action potential, spontaneous activity, or the resting conductance but blocked electrotonic coupling between LC neurons in neonates, blocked synchronous activity in LC neurons from adult animals, and shifted the reversal potential of the opioid current to the potassium equilibrium potential. 8. All results are consistent with the hypothesis that LC cells are electrotonically coupled. Such electrotonic coupling at the level of the nucleus locus coeruleus would mediate synchronous regulation of noradrenaline in widespread areas of the brain. PMID- 7472361 TI - Decreased monosynaptic GABAB-mediated inhibitory postsynaptic potentials in hippocampal CA1 pyramidal cells in the aged rat: pharmacological characterization and possible mechanisms. AB - 1. gamma-Aminobutyric acid (GABA)-mediated inhibitory postsynaptic potentials (IPSPs) were compared in young and aged rats in CA1 area of the rat hippocampus, with the use of the in vitro intracellular recording technique. D-2-Amino-5 phosphonovaleric acid (APV) and 6-cyano-7-nitroquinoxaline-2,3-dione (CNQX) were used to suppress synaptic potentials mediated by the excitatory amino acids. 2. Under these conditions, stimulation of the stratum radiatum elicited a monosynaptic fast GABAA (fIPSP) and a slow GABAB (sIPSP)-mediated IPSP. The fIPSP and the sIPSP were further isolated in the presence of the GABAB antagonist CGP 35348 or the GABAA antagonists bicuculline or picrotoxin. No age-related changes were observed in the amplitude and the duration of the fIPSP. In contrast, the amplitude (but not the duration) of the sIPSP was significantly reduced in the aged rat. 3. The postsynaptic hyperpolarization and increase in membrane conductance induced in pyramidal cells by bath application of the GABAB agonist baclofen were comparable in both groups of animals, indicating that the postsynaptic GABAB receptors are not altered in the aged rats. 4. Paired-pulse depression of IPSPs was used in young and aged rats to study possible alterations in GABA release or in presynaptic GABAB receptors that control GABA release. When fIPSPs were isolated by bath application of tetrahydro-9-aminoacridine (THA), no significant difference in the magnitude of the paired-pulse depression was observed between young and aged rats. A similar result was found with the paired pulse depression of isolated sIPSPs in the presence of bicuculline or picrotoxin.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7472363 TI - Gustatory neural coding in the monkey cortex: acid stimuli. AB - 1. We sought to define the gustatory neural code for acidic stimuli. Therefore we analyzed the responses of 44 single neurons in the insular cortex of four alert cynomolgus macaques in response to the oral application of four basic taste stimuli (glucose, NaCl, HCl, and quinine HCl) and fruit juice, and to a series of 20 additional acids. 2. Neurons responsive to gustatory stimulation were encountered within a volume of 38.2 mm3 (3.5 mm anteroposterior x 2.1 mm mediolateral x 5.2 mm dorsoventral). Taste cells constituted 81 (5.2%) of the 1,552 neurons whose sensitivities were tested. Of these, the activity of 44 was followed through at least one complete application of the stimulus series, and those responses compose the data of this study. Nongustatory cells included those responsive to mouth movements (36.3%), tactile stimulation within the mouth (2.1%), visual approach of the taste stimulus (1.4%), and extension of the tongue (0.1%). The functions of the remaining 54.8% were not determined. 3. The mean spontaneous discharge rate of these cortical taste cells was 3.0 spikes/s (range 0.0-14.4 spikes/s). The mean breadth of tuning coefficient was a moderate 0.72 (range 0.26-0.98). Most evoked activity was excitatory, although inhibition was a prominent response option for four (9%) taste cells. 4. There was no evidence that taste cells with similar functional characteristics were clustered within the cortex, i.e., there was no apparent topographic organization of taste quality. 5. Thirty-four of the 44 cells were divisible into three functional types on the basis of their response profiles to the four basic stimuli used here.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7472362 TI - Contractile properties of the tongue muscles: effects of hypoglossal nerve and extracellular motoneuron stimulation in rat. AB - 1. Stimulation of the whole hypoglossal (XIIth) nerve or its medial or lateral branch in the rat produced two major movements of the tongue as measured with a single force transducer attached to the tip of the tongue. Stimulation of the whole XIIth nerve or the lateral branch produced a retrusion of the tongue, whereas stimulation of the medial branch produced a protrusion. 2. The average retrusive twitch tension evoked by stimulation of the XIIth nerve (11.25 g) or the lateral branch (12.02 g) was significantly greater (P < or = 0.0001) than the protrusive twitch tension (1.05 g) elicited by medial branch stimulation. The tetanic tension produced by lateral branch stimulation (36.82 g) was significantly greater (P < or = 0.007) than the whole nerve tetanic tension (28.23 g). The greater tension elicited by stimulation of the lateral branch of the nerve when compared with the tension elicited by stimulation of the whole XIIth nerve was probably due to the absence of protrusive axons in the lateral branch of the nerve. Stimulation of the whole XIIth nerve activates axons innervating both protrusive and retrusive muscles, resulting in a weaker net retrusive force. 3. The contraction time of the tongue in response to medial branch stimulation (10.94 ms) was significantly faster than when the whole XIIth nerve (15.68 ms, P < or = 0.007) or lateral branch (13.36 ms, P < or = 0.05) was stimulated. The twitch contraction time of the tongue in response to whole XIIth nerve or lateral branch stimulation was not significantly different.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7472364 TI - Extracellular potassium, volume fraction, and tortuosity in rat hippocampal CA1, CA3, and cortical slices during ischemia. AB - 1. An in vitro slice model of ischemia was used to study changes in extracellular potassium concentration and diffusion properties in the stratum pyramidale of CA1 and CA3 regions of the hippocampus and in the cortex of the rat. Slices were submerged in artificial cerebrospinal fluid, and ischemia was induced by removing oxygen and glucose until anoxic depolarization occurred. 2. Extracellular potassium concentration was measured with a valinomycin-based ion-selective microelectrode. The bathing medium contained 5 mM potassium, and in vitro ischemia caused the potassium concentration to rise to 45 mM in CA1, 12 mM in CA3, and 32 mM in cortex. 3. Extracellular volume fraction and tortuosity were determined during normoxic conditions and in vitro ischemia by measuring the diffusion of tetramethylammonium. This cation was iontophoretically released into the extracellular space and its concentration as a function of time determined with an ion-selective microelectrode approximately 100 microns away from the source. 4. During normoxia the volume fraction was 0.14, 0.20, and 0.18, and tortuosity was 1.50, 1.57, and 1.62 in CA1, CA3, and cortex, respectively. These data confirm that the volume fraction of CA1 is smaller than in the two other regions. 5. During ischemia the volume fraction decreased to 0.05, 0.17, and 0.09 in CA1, CA3, and cortex, respectively. Only in CA3 did the tortuosity change significantly by increasing to 1.75. Because of limitations in the time resolution of the diffusion method, the changes in volume fraction and tortuosity during the anoxic depolarization phase of ischemia may have been underestimated.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7472365 TI - Extracellular inhibition and intracellular enhancement of Ca2+ currents by Pb2+ in bovine adrenal chromaffin cells. AB - 1. Effects of highly neurotoxic, inorganic lead ions (Pb2+) on voltage-dependent calcium channels were investigated with the use of the whole cell patch-clamp technique in bovine adrenal chromaffin cells maintained in short-term primary culture (1-5 days). 2. Extracellularly applied Pb2+ induced a concentration dependent, reversible inhibition of Ca2+ currents, with an estimated IC50 approximately equal to 3.0 x 10(-7) M free Pb2+. 3. Elevation of the intracellular free Ca2+ concentration above 10(-8) M dose-dependently reduced the amplitude of the initial Ca2+ current and increased the exponential rate of current rundown. 4. Intracellularly applied Pb2+ prevented the Ca(2+)-dependent reduction of the initial Ca2+ current amplitude and altered the current rundown kinetics from exponential to linear. The effect was dose dependent and saturable, with an estimated EC50 approximately equal to 2.0 x 10(-10) M free Pb2+. 5. These results indicate that in contrast to extracellular blockade, intracellular Pb2+ promotes Ca2+ currents by attenuating the Ca(2+)-dependent, steady-state inactivation of calcium channels. This provides a novel mechanism through which Pb2+ may disrupt calcium signaling in chronically lead-exposed cells. PMID- 7472366 TI - Involvement of cochlear efferent pathways in protective effects elicited with binaural loud sound exposure in cats. AB - 1. Studies in guinea pigs have shown that the crossed olivocochlear efferent [crossed olivocochlear bundle (OCB)] pathways can reduce the cochlear neural desensitization caused by loud sounds. In one experimental paradigm, binaural loud sound exposure produced less damage than did monaural exposure, and various tests confirmed that this could be attributed to the OCB. In contrast, a study in cats has shown no such protection from the OCB for binaural exposures. There are some methodological differences between the cat and guinea pig studies that could account for the difference. The present study was carried out to determine whether two factors, namely anesthetic or exposure frequency, could account for the difference. Experiments were carried out in cats using barbiturate anesthesia, as in the guinea pig experiments. 2. Using a unilateral middle ear muscle (MEM) tenotomy, it was confirmed that under Nembutal anesthesia the MEM did not affect the threshold losses to monaural or binaural exposure. However, comparing results for monaural versus binaural 11-kHz exposures, there were significantly less threshold losses to the binaural exposure, independent of the presence of the MEM. No such difference between monaural and binaural exposures was seen for 3-kHz exposures. 3. By employing unilateral surgical strategies such as MEM tenotomy and/or various brain stem incisions, it was confirmed that the protection with binaural compared with monaural 11-kHz exposure was due specifically to the OCB. In unilaterally deefferented animals, binaural 11-kHz exposure always produced lower threshold losses on the OCB-intact side than on the OCB-cut side, regardless of the status of the MEM. Brain stem cuts that affected other rostrally or laterally located structures but not the OCB produced similar threshold losses bilaterally after binaural exposure, and the losses were comparable with the protected levels seen in other cases with intact OCB and binaural exposure. These data confirmed that when using an exposure frequency very similar to that used in the guinea pig experiments, a protective OCB effect could be demonstrated in cats, as previously seen in guinea pigs. The MEMs appeared to be inactive in barbiturate-anesthetized animals and were not activated by 3-, 7-, or 11-kHz exposures at 100 dB SPL for 10 min. The companion manuscript demonstrates this same effect across a wider range of exposures.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 7472367 TI - Frequency and loss dependence of the protective effects of the olivocochlear pathways in cats. AB - 1. In the previous manuscript I suggested a frequency dependency to olivocochlear bundle (OCB)-mediated protection from loud sound by showing protection for binaural compared with monaural 11-kHz exposures but not 3-kHz exposures of the same intensity and duration. To determine whether this was the case, experiments were carried out in barbiturate-anesthetized cats using the paradigm of a unilateral brain stem incision to deefferent one cochlea in each animal before presentation of a binaural loud sound exposure. With equal-intensity, equal duration binaural exposures, in different groups protection in OCB-intact compared with OCB-cut ears was seen only for exposures at 11, 15, or 20 kHz, but not at 3 or 7 kHz, suggesting that OCB-mediated protection was found only for higher-frequency exposures. This would be consistent with the OCB-mediated protection in guinea pig studies where 10-kHz exposures were used and its absence in a study in cats where 6-kHz exposures were used. However, this conclusion had to be qualified by the fact that the lower-frequency exposures resulted in smaller threshold losses than did the higher-frequency exposures. 2. To determine whether OCB-mediated protection could be obtained for lower-frequency exposures that were made as damaging as or more damaging than the high-frequency exposures, longer-duration, lower-frequency exposures were used. OCB-mediated protection could then be obtained for exposure at 7 kHz, 100 dB SPL for 15 min but not at 3 or 5 kHz, 100 dB SPL for 20 min or at 3 kHz, 100 dB SPL for 40 min or 106 dB SPL for 20 min. Finally, when large threshold losses were produced with exposure at 3 kHz, 106 dB SPL for 40 min, OCB-mediated protection could be obtained for this low-frequency exposure too. These effects suggested that there were different "activation threshold" for OCB-mediated protection as a function of exposure frequency. To determine whether this also applied for the higher-frequency exposures (11, 15, and 20 kHz), all of which had elicited OCB-mediated protection when presented at 100 dB SPL for 10 min, these exposure frequencies were presented at 100 dB SPL for 7 min to produce low threshold losses. Now protection was found for the 11- and 15-kHz exposures but not for the 20-kHz exposure. 3. Thus the activation threshold for OCB-mediated protection varied in a frequency dependent manner.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 7472368 TI - Serial E-M and simulation study of presynaptic inhibition along a group Ia collateral in the spinal cord. AB - 1. A muscle spindle primary afferent (group Ia) was physiologically identified and labeled intracellularly with the use of horseradish peroxidase (HRP) in the cat lumbar spinal cord. Serial-section electron microscopy (EM) was used to examine and reconstruct an entire axon collateral and its branches within Clarke's column. In the present study the existence and location of presynaptic contacts on Ia afferent boutons along these collateral branches were determined from examination of the serial-section electron-micrographs. 2. Of 36 Ia boutons examined in serial sections along the branches of the same collateral, 3 presynaptic contacts were found. Two of these contacts were made with Ia boutons in a complex nodal region consisting of two unmyelinated side branches exhibiting a total of six Ia boutons. The other presynaptic contact was made with a Ia bouton in a nodal region consisting of two Ia boutons connected by a thin unmyelinated bridge. 3. Computer simulations, based directly on the serial section-reconstructions, were used to investigate the possible effects of these presynaptic contacts on membrane potential and on a propagating action potential along the Ia collateral. The effect of a presynaptic contact was modeled by a sustained gamma-aminobutyric acid-A (GABAA)-activated chloride conductance. 4. The simulation results indicated that the effect of a presynaptic contact on membrane potential and action-potential amplitude is likely to extend beyond the contacted bouton to other boutons occurring along the short unmyelinated branches arising from the same node.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7472369 TI - Modulation of high-frequency vestibuloocular reflex during visual tracking in humans. AB - 1. Humans may visually track a moving object either when they are stationary or in motion. To investigate visual-vestibular interaction during both conditions, we compared horizontal smooth pursuit (SP) and active combined eye-head tracking (CEHT) of a target moving sinusoidally at 0.4 Hz in four normal subjects while the subjects were either stationary or vibrated in yaw at 2.8 Hz. We also measured the visually enhanced vestibuloocular reflex (VVOR) during vibration in yaw at 2.8 Hz over a peak head velocity range of 5-40 degrees/s. 2. We found that the gain of the VVOR at 2.8 Hz increased in all four subjects as peak head velocity increased (P < 0.001), with minimal phase changes, such that mean retinal image slip was held below 5 degrees/s. However, no corresponding modulation in vestibuloocular reflex gain occurred with increasing peak head velocity during a control condition when subjects were rotated in darkness. 3. During both horizontal SP and CEHT, tracking gains were similar, and the mean slip speed of the target's image on the retina was held below 5.5 degrees/s whether subjects were stationary or being vibrated at 2.8 Hz. During both horizontal SP and CEHT of target motion at 0.4 Hz, while subjects were vibrated in yaw, VVOR gain for the 2.8-Hz head rotations was similar to or higher than that achieved during fixation of a stationary target. This is in contrast to the decrease of VVOR gain that is reported while stationary subjects perform CEHT.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7472370 TI - Pharmacological characterization of extracellular pH transients evoked by selective synaptic and exogenous activation of AMPA, NMDA, and GABAA receptors in the rat hippocampal slice. AB - 1. Inhibitors of extracellular carbonic anhydrase (CAo) offer much promise as diagnostic tools in the study of the synaptic basis of activity-induced alkaline transients in the brain. However, most of the present information related to the effects of CAo blockers in nervous tissue comes from experiments that involve simultaneous synaptic activation of various types of postsynaptic receptor channels. In the present work, double-barreled H(+)-selective microelectrodes were used to study alkaline shifts in extracellular pH (pHo) evoked by selective synaptic and pharmacological activation of glutamate and gamma-aminobutyric acid (GABA) receptors in the CA1 cell body layer in rat hippocampal slices. Inhibition of CAo was achieved with the use of the poorly permeant carbonic anhydrase inhibitor, benzolamide (10 microM; applied in the bath solution) or the impermeant macromolecular inhibitor, prontosil-dextran 5000 (PD 5000; applied in microdrops). 2. Alkaline transients that were exclusively attributable to synaptic activation of glutamate receptors were induced by stimulation of Schaffer collaterals in the presence of picrotoxin (PiTX, 100 microM). An enhancement by the CAo inhibitors of these alkaline transients took place at all stimulus frequencies (5-200 Hz) and stimulus train durations (0.5-20 s) examined. 3. Inhibition of CAo enhanced the alkaline transients evoked by selective synaptic activation of alpha-amino-3-hydroxy-5-methyli-oxazolate- 4-propionic acid (AMPA)/kainate receptors in experiments involving stimulation of Schaffer collaterals in the simultaneous presence of PiTX and D-2-amino-5 phosphonopentoate (AP5, 40-80 microM). 4. Alkaline shifts evoked by selective synaptic activation of N-methyl-D-aspartate (NMDA) receptors were enhanced after inhibition of CAo as seen in experiments where Schaffer collaterals were stimulated in the simultaneous presence of PiTX and 6-cyano-2,3-dihydroxy-7 nitroquinoxaline (CNQX, 20-40 microM) in an Mg(2+)-free solution. 5. Benzolamide and PD 5000 also enhanced the alkaline shifts seen upon pressure injection of glutamate, AMPA, or NMDA. The glutamate-induced alkaline shifts were inhibited by AP5+CNQX, suggesting that uptake of glutamate did not significantly contribute to their generation. 6. Stimuli applied at 5-10 Hz in stratum radiatum close (within 0.5 mm) to the recording site evoked alkaline shifts that were blocked by CNQX plus AP5. In the continuous presence of the two glutamate antagonists, PiTX sensitive alkaline transients were observed in response to brief high-frequency (20-100 Hz) trains consisting of 100 stimuli. Upon application of pentobarbital sodium (100 microM), these apparently monosynaptically evoked GABAA receptor mediated alkaline transients were evident also at low stimulation frequencies (5 10 Hz).(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 7472371 TI - Relative contributions of excitatory and inhibitory neuronal activity to alkaline transients evoked by stimulation of Schaffer collaterals in the rat hippocampal slice. AB - 1. The neuronal basis of alkaline shifts in extracellular pH (pHo) evoked by stimulation of Schaffer collaterals was studied by means of double-barreled H(+) selective microelectrodes in the area CA1 of rat hippocampal slices. 2. Alkaline transients in stratum pyramidale evoked by stimulation at a low frequency (5-10 Hz) were enhanced by pentobarbital sodium (100 microM). In the absence of the drug, inhibition of extracellular carbonic anhydrase (CAo) by benzolamide or by prontosildextran 5000 (PD 5000) resulted in an increase in the alkaline shifts. In contrast to this, alkaloses evoked by low-frequency stimulation in the presence of pentobarbital were attenuated by a subsequent inhibition of CAo. 3. Blockade of gamma-aminobutyric acid-A (GABAA) receptors with picrotoxin (PiTX; 100 microM) resulted in an enhancement of alkaline transients in s. pyramidale evoked by low-frequency stimulation (10 Hz) but suppressed alkaline shifts evoked by brief high-frequency (1 s, 100 Hz) trains of stimuli. 4. Application of trains of stimuli consisting of a constant number of pulses (50 or 100) revealed a striking dependence of the effect of benzolamide on stimulation frequency (10-200 Hz) in s. pyramidale: the enhancement of the alkaloses seen upon inhibition of CAo became progressively smaller with an increase in frequency, and at 100-200 Hz benzolamide produced a suppression or a complete block of the alkaline transients. However, alkaline transients evoked with the use of a constant train duration (5 s) were enhanced by benzolamide at all stimulation frequencies examined (5-200 Hz).(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7472372 TI - Diverse neuronal populations mediate local circuit excitation in area CA3 of developing hippocampus. AB - 1. Studies were undertaken to better understand why the developing hippocampus has a marked capacity to generate prolonged synchronized discharges when exposed to gamma-aminobutyric acid-A (GABAA) receptor antagonists. 2. Excitatory synaptic interactions were studied in small microdissected segments of hippocampal area CA3. Slices were obtained from 10- to 16-day-old rats. Application of the GABAA receptor antagonist penicillin produced prolonged synchronized discharges in minislices that were very similar, if not identical, to those recorded in intact slices. The sizes of minislices were systematically varied. Greater than 90% of those that measured 600 microns along the cell body layer produced prolonged synchronized discharges, whereas most minislices measuring 300 microns produced only brief interictal spikes. 3. Action potentials in the majority (75%, 158 of 254) of cells impaled with microelectrodes were able to entrain the entire CA3 population. They were also able to increase (on average 26%) the frequency of spontaneous population discharges. The population discharges were followed by a refractory period that lasted 5-60 s, during which single cells were unable to initiate a population discharge. 4. The majority (87%) of neurons with intrinsic burst properties were found to entrain the CA3 population. The electrophysiological characteristics of these cells were reminiscent of recordings obtained from more mature rats. Action potentials were quite prolonged and demonstrated a secondary shoulder or hump on the down-slope of the spike. 5. When bursting cells were filled with Lucifer yellow and imaged during recording sessions by videomicroscopy and later using confocal microscopy, they showed the anatomic features of CA3 hippocampal pyramidal cells. Confocal microscopy permitted detailed characterization of individual neurons and showed substantial variation in cellular microanatomy. 6. Another class of cells that were found to entrain the CA3 population but did not demonstrate intrinsic bursts were termed regular-firing cells. These cells possessed many of the anatomic and physiological features of bursting cells with the exception of burst firing. They were rarely encountered in intracellular recordings. 7. The third physiological class of cells was termed fast-spiking cells. These had action potentials that were shorter in duration than the other two cell types. They were distinct in the rapid rate of spike repolarization. They demonstrated modest degrees of spike frequency adaptation and fired repeatedly and at relatively high frequencies. Compared with reports on fast-spiking cells in mature hippocampus and neocortex, action potentials appear to be slower and repetitive discharging appeared to be of a lower frequency.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 7472373 TI - Enhancement of GABAA receptor-mediated conductances induced by nerve injury in a subclass of sensory neurons. AB - 1. The effects of axotomy on the electrophysiologic properties of adult rat dorsal root ganglion (DRG) neurons were studied to understand the changes in excitability induced by traumatic nerve injury. Nerve injury was induced in vivo by sciatic nerve ligation with distal nerve transection. Two to four weeks after nerve ligation, a time when a neuroma forms, lumbar (L4 and L5) DRG neurons were removed and placed in short-term tissue culture. Whole cell patch-clamp recordings were made 5-24 h after plating. 2. DRG neurons were grouped into large (43-65 microns)-, medium (34-42 microns)-, and small (20-32 microns)- sized classes. Large neurons had short duration action potentials with approximately 60% having inflections on the falling phase of their action potentials. In contrast, action potentials of medium and small neurons were longer in duration and approximately 68% had inflections. 3. Pressure microejection of gamma aminobutyric acid (GABA, 100 microM) or muscimol (100 microM) onto voltage clamped DRG neurons elicited a rapidly desensitizing inward current that was blocked by 200 microM bicuculline. To measure the peak conductance induced by GABA or muscimol, neurons were voltage-clamped at a holding potential of -60 mV, and pulses to -80 mV and -100 mV were applied at a rate of 2.5 or 5 Hz during drug application. Slope conductances were calculated from plots of whole cell current measured at each of these potentials. 4. GABA-induced currents and conductances of control DRG neurons increased progressively with cell diameter. The mean GABA conductance was 36 +/- 10 nS (mean +/- SE) in small neurons, 124 +/ 21 nS in medium neurons, and 527 +/- 65 nS in large neurons. 5. After axotomy, medium neurons had significantly larger GABA-induced conductances compared with medium control neurons (390 +/- 50 vs. 124 +/- 21; P < 0.001). The increase in GABA conductance of medium neurons was associated with a decrease in duration of action potentials. In contrast, small neurons had no change in GABA conductance or action potential duration after ligation. The GABA conductance of large control neurons was highly variable, and ligation resulted in an increase that was significant only for neurons > 50 microns. The mean action potential duration in large neurons was not significantly changed, but neurons with inflections on the falling phase of the action potential were less common after ligation. There was no difference in resting potential or input resistance between control and ligated groups, except that the resting potential was less negative in small cells after axotomy.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 7472374 TI - Directional variation of spatial and temporal characteristics of limb movements made by monkeys in a two-dimensional work space. AB - 1. The directional variation of kinematic and electromyographic (EMG) characteristics of two-joint arm movements made to targets in a two-dimensional work space was studied in monkeys trained to make targeted arm movements under different behavioral conditions. 2. In each animal, kinematic measures of movement (movement amplitude, movement time, peak velocity, and trajectory curvature) and endpoint spatial position within the target zone varied as a function of the direction of the target from the starting position. Movements made toward the body into the ipsilateral hemispace generally had the smallest amplitude, lowest peak velocity, and longest movement time. 3. Although the directional variation in peak velocity could partially be accounted for by predicted anisotropies in the inertial load imposed by the arm, deviations from these predictions suggest that movement amplitude is controlled more rigorously by the CNS. Adjustments in movement time may be used to compensate for inertial anisotropies. 4. The spatial characteristics of movements (amplitude, trajectory curvature, or endpoint error) were influenced little by the visibility of the target during movement, the advanced knowledge of target location, or the presence or absence of an external trigger cue. However, temporal characteristics (movement time, peak velocity, and for some animals, reaction time) varied more as sensory cues were changed. 5. The time of initial EMG activity in muscles acting around the shoulder varied systematically as a function of target direction. A cosine model accounted for a large fraction of the variability in initial onset time, as determined in a trial-by-trial analysis. The amplitude of the EMG activity was more narrowly tuned, however. Muscles acting at the elbow showed less activity and more variable directional tuning. 6. We conclude that directional variations in the kinematic characteristics of movement, and thus, the dynamic force requirements of the task, must be taken into consideration as contributors to the apparent directional coding described for neuronal populations in different portions of the CNS. PMID- 7472375 TI - Covert orienting of attention in macaques. II. Contributions of parietal cortex. AB - 1. To understand some of the contributions of parietal cortex to the dynamics of visual spatial attention, we recorded from cortical cells of monkeys performing attentional tasks. We studied 484 neurons in the intraparietal sulcus and adjacent gyral tissue of two monkeys. We measured phasic responses to peripheral visual stimuli while the monkeys attended toward or away from the stimuli or when attention was not controlled. Neurons were tested while the monkeys gazed at a spot of light (simple fixation task), actively attended to a foveal target (foveal attention task), performed a reaction time task (cued reaction time task), made saccadic eye movements to visual targets (saccade task), or responded to a repetitious peripheral target (probability task). 2. In a previous paper we demonstrated that monkeys, like humans, responded more quickly to visual targets when the targets followed briefly flashed visual cues (validly cued targets) (Bowman et al. 1993). It has been hypothesized that the cue attracts attention to its locus and results in faster reaction times (Posner 1980). In the present physiological studies, visual cues consistently excited these neurons when they were flashed in the receptive field. Such activity might signal a shift of attention. Visual targets that fell within the receptive field and that immediately followed the cue evoked relatively weak responses. This response was due to a relative refractory period. 3. Next we tested attentional processes in these tasks that were independent of the visual response to the cue. We placed the cue outside of the receptive field and the target within the receptive field. We found that 23% of these cells had a significant decrease in their firing rate to validly cued targets in their receptive fields under these conditions. Strong responses were evoked by the same target when the cue was flashed in the opposite hemifield (invalidly cued targets). Thus this group of neurons responded best when attention was directed toward the opposite hemifield. 4. For another group of parietal cells (13%) there was an enhanced response to targets in the visual receptive field when the cue was in the same hemifield. For the remaining 64% of the cells there was no significant modulation in this task. 5. The cued reaction time task involved exogenous control of attention; the sensory cue gave spatial and temporal direction to attention. We used several other tasks to test for endogenous control of attention.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 7472376 TI - Covert orienting of attention in macaques. III. Contributions of the superior colliculus. AB - 1. The present experiments were conducted to study physiological mechanisms in the superior colliculus and their relation to visual spatial attention. We used a cued reaction time task studied in detail previously (Bowman et al. 1993; Posner 1980). Monkeys learned to fixate a spot of light and release a bar when a target light appeared. Cues on the same side as the target (valid cue) were associated with faster reaction times than those on the opposite side (invalid cue). The difference in reaction times is hypothesized to be a measure of attention. 2. A total of 79 neurons within the superficial layers of the superior colliculi of two monkeys were studied. When the cues and targets were positioned so that both were within the visual receptive field, the cues excited the cells, and this produced a refractoriness to the targets for the following 400 ms. Both the ON and OFF responses to the cue were constant under all conditions. 3. These neurons were also tested with the cue just outside of the visual receptive field. This was done to avoid refractory effects from the cue; there was no significant modulation of the response to the target under these conditions. The visual responses of neurons in the intermediate layers of the superior colliculus also responded equivalently under these conditions. 4. When the activity of cells within the foveal representation was compared during the performance of three tasks, there was differential activity. The appearance of the fixation point during the performance of the cued reaction time task led to a strong, transient discharge.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7472377 TI - Electrophysiological evidence for overlapping dominant and latent inputs to somatosensory cortex in squirrel monkeys. AB - 1. The pattern of reorganization in area 3b of adult primates after median or ulnar nerve section suggests that somatic afferents from the dorsum of the hand, carried by the radial nerve, have preferential access to the cortical territories normally expressing glabrous inputs carried by the median and ulnar nerves. A likely mechanism underlying preferential access is preexisting, but silent, radial nerve inputs to the glabrous region of cortex. 2. We tested this by comparing the effects of electrical stimulation of median or ulnar versus radial nerves, on responses in the hand representation of area 3b. Laminar current source density and multiunit activity profiles were sampled with the use of linear array multicontact electrodes spanning the laminae of area 3b. Data were obtained from three squirrel monkeys anesthetized during recording. 3. Compared with colocated median or ulnar nerve responses, the radial nerve response had 1) an initial short-latency response in the middle laminae that was subtle; there was a small transmembrane current flow component without a discernable multiunit activity correlate; and 2) a laminar sequence and distribution of activity that was similar to those of the median or ulnar nerve responses (i.e., initial activation of the middle, followed by upper and lower laminae), but the significant current flow and multiunit response to radial nerve stimulation occurs 12-15 ms later. 4. Normal corepresentation of nondominant dorsum hand (radial) inputs with the dominant (median or ulnar) inputs in the glabrous hand surface representation provides a clear vehicle for the biased patterns of reorganization occurring after peripheral nerve section. The initial, "subtle" activity phase in the nondominant response is believed to reflect intracortical inhibition, and the later "significant" response phase, a rebound excitation, possibly compounded by an indirect or extralemniscal input. The spatiotemporal pattern of nondominant input is proposed to play a role in normal somatosensory perception. PMID- 7472378 TI - Information coding in the rodent prefrontal cortex. I. Single-neuron activity in orbitofrontal cortex compared with that in pyriform cortex. AB - 1. Extracellular spike activity was recorded from 1,942 single neurons in orbitofrontal cortex (OF) and 591 single neurons in pyriform cortex (PIR) over multiple sessions in rats performing an eight-odor discrimination task in which the stimulus sequence contained predictable associations between particular odor pairs. Neural firing patterns were examined in relation to task events in the current trial and variables associated with current sensory processing, events of recent past trials, and long-term associations involving the odor cues. 2. Overall, 34% of single neurons in OF and 30% of single neurons in PIR fired selectively during one or more salient trial events including trial initiation, odor sampling, performance of the discriminative response, and water consumption. The activity of other cells recorded in OF (13%) and PIR (10%) was suppressed for the duration of each trial. Although the proportion of some cell types differed between the two areas, the firing patterns of OF and PIR neurons were qualitatively indistinguishable. 3. Firing during odor sampling and the discriminative response was influenced by the identity of the current odor. Some cells fired selectively to a single odor, but most cells were coarsely tuned such that they fired to several of the eight odors to differing degrees consistent with previous reports. Considerable odor coding was observed in both OF and PIR. 4. Firing during trial initiation and odor sampling was also influenced by the identity and reward association of the odor presented in the immediately preceding trial. The influence of past odor identity and valence was observed in both OF and PIR. 5. Firing during trial events was also influenced by the acquired associations between odors and their assigned reward contingencies and between pairs of odors involved in predictive relationships. The reward valence of the current odor significantly influenced firing during odor sampling and the discriminative response; some cells responded preferentially to rewarded odors and others to nonrewarded odors. Firing during trial initiation and odor sampling reflected whether or not the odor in the current trial had been predicted by the odor in the preceding trial. In addition, firing during odor sampling reflected the expectation of reward in the following trial that could be inferred from the predictable associations between odors. Each of these properties was observed in both OF and PIR. 6. The findings in OF were consistent with the view that prefrontal subdivisions mediate the temporal organization of complex behaviors within specific informational domains. OF appears to be concerned with the specific domain of olfaction.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 7472379 TI - Information coding in the rodent prefrontal cortex. II. Ensemble activity in orbitofrontal cortex. AB - 1. Neural activity was recorded from the orbitofrontal cortex (OF) of rats performing an eight-odor discrimination task that included predictable associations between particular odor pairs. A modified linear discriminant analysis was employed to characterize the population response in each trial of the task as a point in an N-dimensional activity space with the firing rate of each cell in the population represented on one of the N dimensions. The ability of the ensemble to discriminate among conditions of a variable was reflected in the tendency of population responses to cluster together in this activity space for repetitions of a given condition. We assessed coding of several variables describing the period of odor sampling, focusing on aspects of current, past, and future events reflected in single-neuron firing patterns, in ensembles composed of 22-138 cells active during the period when the rats sampled the discriminative stimulus in each trial. 2. OF ensembles performed well at discriminating variables with relevance to task demands represented in single-neuron firing patterns, specifically the physical attributes and assigned reward contingency of the current odor as well as the expectation of reward in the following trial that could be inferred from the predictable associations between particular pairs of odors. OF ensembles were able to correctly identify the identity and assigned reward contingency of the current odor in up to 52% (chance = 12.5%) and 99% (chance = 50%) of all trials, respectively, such that the observed behavioral performance required a population of 5,364 odor-responsive cells in the case of odor identity and only 40 cells in the case of valence. Expectations regarding upcoming rewards based on both assigned response contingency and associations between particular pairs of odors were correctly classified in up to 67% (chance = 20%) of all trials such that the observed level of behavioral performance required a population of 3,169 cells. 3. Other information represented in the single-neuron firing patterns, such as the identity and reward contingency of the preceding odor and specific odor-odor associations, was poorly encoded by OF ensembles. Thus neural ensembles in OF may represent only some of the information reflected in single-neuron activity. Stable coding of only the most useful and relevant information by the ensemble might emerge from the tuning properties of single neurons under the influence of the task at hand, producing in the well trained animal the observed pattern of broad and diverse coding by single neurons and selective, task-relevant coding by neural ensembles in OF. PMID- 7472380 TI - Low-threshold neuronal activity of spinal dorsal horn neurons increases during REM sleep in cats: comparison with effects of anesthesia. AB - 1. Cats were prepared for chronic recordings from the lumbar enlargement of the spinal dorsal horn. At the beginning of each recording session, a tungsten microelectrode was advanced through the dura in a physiologically intact, awake, drug-free animal, until amplitude discrimination provided a single neuron with a receptive field on the hindquarters. 2. Extracellular recordings of activity of each neuron were made during receptive field stimulation with tactile and thermal nonnoxious and noxious stimuli. 3. Baseline responses obtained in the awake state were compared with responses of the same neurons during slow-wave or rapid-eye movement (REM) sleep. In a subpopulation of neurons, the effects of anesthesia (propofol, 7.5 mg/kg iv) were observed after the completion of sleep studies. 4. The low-threshold receptive fields of the seven neurons studied during REM sleep were all increased in size when compared with the baseline value. The average increase was 52.6% (range 26.2-96.7%). 5. The low-threshold receptive fields of the seven neurons studied during REM sleep were reduced by propofol anesthesia by an average of 49.1% (range 29-74%). 6. Neuronal response to receptive field brushing was observed in 15 neurons during REM sleep. The effect of propofol on receptive field brushing was observed in 8 of those neurons. In only one of those eight neurons were the effects of REM sleep and anesthesia in the same direction. 7. Changes in neuronal responses were less consistent during slow-wave sleep but still differed from changes induced by propofol.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7472381 TI - Involvement of NMDA receptors in the respiratory phase transition is different in the adult guinea pig in vivo and in the isolated brain stem preparation. AB - 1. We investigated the involvement of N-methyl-D-aspartate (NMDA) receptors in the respiratory pattern in an in vitro preparation of adult brain stem compared with in vivo conditions in the guinea pig. 2. In vivo, combining administration of the NMDA channel blocker dizocilpine (MK-801) (3 mg/kg) with a surgical section of the vagus nerves induced an apneustic type of respiration characterized by long inspiratory "holds," as has been shown in other species. The same effect was observed in hypothermic animals (30 degrees C). 3. The isolated in vitro brain stems from these apneustic animals did not present a prolonged inspiratory phase. A second dose of dizocilpine (100 microM perfused vascularly did not induce apneusis, even after increasing brain stem temperature to 35.5 degrees C. 4. In another group of isolated brain stems of adult guinea pigs anesthetized with pentobarbital sodium before decapitation, we perfused dizocilpine and NMDA through the basilar artery. The duration of periodic inspiratory motor activity recorded from the hypoglossal nerve was unaffected by dizocilpine (1-100 microM) or the competitive NMDA antagonist D- or DL-2-amino-5 phosphonopentanoic acid (100 microM and 1 mM), although respiratory frequency decreased. The increase in respiratory activity produced by vascularly perfused NMDA (25-100 microM) was blocked by dizocilpine (100 microM). 5. We conclude that the central mechanism of inspiratory termination in the vagotomized adult guinea pig requires the activation of NMDA receptors in vivo but not in vitro. This difference is not due to the hypothermic environment in vitro. Possible mechanisms for phase switching in vitro are discussed. PMID- 7472382 TI - Receptive field expansion in adult visual cortex is linked to dynamic changes in strength of cortical connections. AB - 1. Receptive field (RF) sizes of neurons in adult primary visual cortex are dynamic, expanding and contracting in response to alternate stimulation outside and within the RF over periods ranging from seconds to minutes. The substrate for this dynamic expansion was shown to lie in cortex, as opposed to subcortical parts of the visual pathway. The present study was designed to examine changes in cortical connection strengths that could underlie this observed plasticity by measuring the changes in cross-correlation histograms between pairs of primary visual cortex neurons that are induced to dynamically change their RF sizes. 2. Visually driven neural activity was recorded from single units in the superficial layers of primary visual cortex in adult cats, with two independent electrodes separated by 0.1-5 mm at their tips, and cross-correlated on-line. The neurons were then conditioned by stimulation with an "artificial scotoma," a field of flashing random dots filling the region of visual space around a blank rectangle enclosing the RFs of the recorded neurons. The neuronal RFs were tested for expansion and their visually driven output again cross-correlated. After this, the neurons were stimulated vigorously through their RF centers to induce the field to collapse, and the visually driven output from the collapsed RFs was again cross-correlated. Cross-correlograms obtained before and after conditioning, and after RF collapse, were normalized by their flanks to control for changes in peak size due solely to fluctuations in spike rate. 3. A total of 37 pairs of neurons that showed distinct cross-correlogram peaks, and whose RF borders were clearly discernible both before and after conditioning, were used in the final analysis. Of these neuron pairs, conditioning led to a clear expansion of RF boundaries in 28 pairs, whereas in 9 pairs the RFs did not expand. RFs that did expand showed no significant shifts in their orientation preference, orientation selectivity, or ocularity. 4. When the RFs of a pair of neurons expanded with conditioning, the area of the associated flank-normalized cross correlogram peaks also increased (by a factor ranging from 0.84 up to 3.5). Correlograms returned to their preconditioning values when RFs collapsed.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 7472383 TI - Effective synaptic current and motoneuron firing rate modulation. AB - 1. We used a modified voltage-clamp technique to measure the steady-state effective synaptic currents (I(N)) produced by activating four different input systems to cat hindlimb motoneurons: Ia afferent fibers, Ia-inhibitory interneurons, Renshaw interneurons, and contralateral rubrospinal neurons. In the same motoneurons, we measured the slope of the firing rate-injected current (f-I) relation in the primary range. We then reactivated these synaptic inputs during steady, repetitive firing to assess their effects on motoneuron discharge rate. 2. Our measurements of I(N) were derived from recordings made near the resting membrane potential, whereas the effects of the synaptic inputs on repetitive discharge were evaluated at more depolarized membrane potentials. Thus we adjusted the I(N) values for these changes in driving force based on estimates of the synaptic reversal potential and the mean membrane potential during repetitive discharge. 3. We found that changes in the steady-state discharge rate of a motoneuron produced by these synaptic inputs could be reasonably well predicted by the product of the estimated value of I(N) during repetitive firing and the slope of the motoneuron's f-I relation. Although there was a high correlation between predicted and observed changes in firing rate for our entire sample of motoneurons (r = 0.93; P < 0.001), the slope of the relation between predicted and observed firing rate modulation was significantly greater than 1. 4. The systematic difference between predicted and observed firing rate modulation observed in the overall sample was primarily due to the fact that our predictions underestimated the changes in firing rate produced by Ia excitation and Ia inhibition.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7472384 TI - Relation between cerebral activity and force in the motor areas of the human brain. AB - 1. Positron emission tomography (PET) studies were performed in six normal right handed male volunteers (age 30 +/- 3) to investigate the relationship between cerebral activation as measured by relative regional cerebral blood flow (rCBF) and force peak exerted during right index finger flexion. The purpose was to determine in which central motor structures activity is directly correlated with force for repeatedly executed movements. 2. Twelve PET rCBF measurements were performed in each volunteer with the use of H2(15)O as a perfusion tracer. Volunteers pressed a Morse-key repetitively with their right index finger for 2 min while lying in a supine position in the PET camera. The device was fitted with strain gauges to measure the force peaks exerted upon it. Scans were collected twice each at five different levels of exerted force peak and in a resting state. Individual and group results were co-registered with anatomic magnetic resonance images (MRI). 3. Group analysis revealed four major regions with a high correlation between rCBF and different degrees of repetitively exerted force peaks. One was located in the arm area of the left lateral surface [primary somatosensory and motor cortex (SI, MI)]. The second area was situated on the left mesial surface of the brain, posterior to the anterior commissure (AC) and encompassing the first gyrus dorsal to the cingulate sulcus. This area is thought to be homologous to the posterior part of the supplementary motor area (SMA) in the monkey. The third area was the dorsal bank of the posterior cingulate sulcus. The fourth area showing a significant correlation between rCBF and force peaks was in the cerebellar vermis. 4. Individual PET data were co registered with each individual's MRI in order to identify precisely the locations of structures demonstrating a positive correlation between rCBF and force peak. Activated areas on the mesial surface consisted of the same two distinct regions seen in the group data. In three subjects the focus on the lateral surface of the cortex appeared to extend into the caudal premotor area; in two it extended into the rostral part of the superior parietal area. In no subject did blood flow in the anterior cingulate areas and anterior SMA show a correlation with the force exerted. Cerebellar correlations were present in the vermis in all subjects.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 7472385 TI - Regional heterogeneity of pathophysiological alterations in CA1 and dentate gyrus in a chronic model of temporal lobe epilepsy. AB - 1. Extracellular and intracellular recording techniques were employed in brain slice preparations to characterize responses of hippocampal tissue in the post self sustaining limbic status epilepticus (post-SSLSE) model of chronic temporal lobe epilepsy (TLE) as compared with responses in slices from control animals. Experiments were performed > or = 1 mo, and up to 7 mo, after status epilepticus. Two regions of the hippocampal formation linked to different aspects of epileptogenesis, the CA1 region and the dentate gyrus (DG), were studied. In any given experiment, CA1 and DG were examined in different slices from the same animal. 2. Pyramidal cells in CA1 were activated by means of electrodes positioned over fiber bundles that monosynaptically project to these cells, either those located in the stratum lacunosum/moleculare or those in the stratum radiatum. Granule cells were similarly activated by electrodes positioned in the perforant path. Full input-output curves were determined by varying stimulus strength and charting the amplitudes of population spikes (PSs). 3. Two indexes, stimulus sensitivity and responsiveness, were quantified in control tissue and in post-SSLSE tissue by means of input-output curves to provide comparisons between normal and epileptic tissue. There were no changes in stimulus sensitivity, defined as the stimulus intensity required to evoke comparable responses in input output curves, between control and post-SSLSE tissue. However, responsiveness, defined as the number of extracellular PSs or intracellular action potentials (APs) elicited by a stimulus strength giving rise to maximal-amplitude PSs, proved a reliable method for identifying and categorizing epileptic responses. This index allowed for comparisons between anatomic regions within an experiment as well as among experiments for the same region. Both CA1 pyramidal cells and DG granule cells from post-SSLSE tissue showed hyperresponsiveness relative to control tissue. 4. Control tissue never exhibited > 2 PSs in either CA1 or DG in response to stimuli that produced maximal-amplitude PSs. Therefore a criterion of > or = 3 PSs was adopted to delineate tissue as hyperresponsive on the basis of extracellular responses. In CA1 about one half of the post-SSLSE slices displayed > or = 3 PSs with stimuli giving maximal-amplitude PSs, meeting the criterion for hyperresponsiveness; in DG about one fifth of the slices showed hyperresponsiveness. 5. CA1 and DG differed with respect to the spectrum of hyperresponsiveness they exhibited, this being more robust in CA1. The two regions studied also showed heterogeneity with respect to maximal PS amplitudes.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 7472386 TI - Changes in inhibitory neurotransmission in the CA1 region and dentate gyrus in a chronic model of temporal lobe epilepsy. AB - 1. In this report we compare changes in inhibitory neurotransmission within the CA1 region and the dentate gyrus (DG) in a model of chronic temporal lobe epilepsy (TLE). Extracellular and intracellular recordings were obtained in combined hippocampal-parahippocampal slices > or = 1 mo after a period of self sustaining limbic status epilepticus (SSLSE) induced by continuous hippocampal stimulation. 2. Polysynaptic inhibitory postsynaptic potentials (IPSPs) were induced by positioning electrodes to activate specific afferent pathways and evoking responses in the absence of glutamate receptor antagonists [D(-)-2-amino 5-phosphonovaleric acid (APV) and 6-cyano-7-nitroquinoxaline-2,3-dione (CNQX)]. Polysynaptic IPSPs were evoked in CA1 pyramidal cells from electrodes positioned in stratum radiatum and in stratum lacunosum/moleculare. Polysynaptic IPSPs were evoked in DG granule cells from electrodes positioned over the perforant path located in the subiculum. Monosynaptic IPSPs were induced by positioning electrodes within 200 microns of the intracellular recording electrode (near site stimulation) and stimulating in the presence of APV and CNQX to block ionotropic glutamate receptors. Monosynaptic IPSPs were evoked in CA1 pyramidal cells with electrodes positioned in the stratum lacunosum/moleculare and stratum pyramidale. Monosynaptic IPSPs were evoked in DG granule cells with electrodes positioned in the stratum moleculare. 3. Population spike (PS) amplitudes were employed to assure that a full range of stimulus strengths, from subthreshold for action potentials to an intensity giving maximal-amplitude PSs, was used to elicit polysynaptic IPSPs in CA1 pyramidal cells in both post-SSLSE and control slices. In control tissue, polysynaptic IPSPs were biphasic, composed of early and late events. In post-SSLSE tissue, polysynaptic IPSPs were markedly diminished. The diminution of polysynaptic IPSPs was detected at all levels of stimulus intensity. Both early IPSPs [mediated by gamma-aminobutyric acid-A (GABAA) receptors] and late IPSPs (mediated by GABAB receptors) were diminished. Polysynaptic IPSPs were diminished with both stratum radiatum and with stratum lacunosum/moleculare stimulation. 4. Reversal potentials for either polysynaptic early or polysynaptic late IPSPs evoked in CA1 pyramidal cells by stratum radiatum stimulation were not different in slices from post-SSLSE animals as compared with control animals. Likewise, reversal potentials for either polysynaptic early or polysynaptic late IPSPs evoked by stratum lacunosum/moleculare stimulation did not differ in the two groups. These findings excluded changes in driving force as an explanation for the diminished amplitude of IPSPs in CA1 pyramidal cells in the post-SSLSE model.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 7472387 TI - Changes in excitatory neurotransmission in the CA1 region and dentate gyrus in a chronic model of temporal lobe epilepsy. AB - 1. In this report we compare changes of excitatory neurotransmission within the CA1 region and the dentate gyrus (DG) in a model of chronic temporal lobe epilepsy (TLE). Extracellular and intracellular recordings were obtained from in vitro hippocampal-parahippocampal slices > or = 1 mo after a period of self sustaining limbic status epilepticus (SSLSE) induced by continuous hippocampal stimulation. Pyramidal cells in CA1 were activated by electrodes in the stratum lacunosum/moleculare or stratum radiatum. Granule cells in DG were similarly activated by electrodes positioned in the perforant path. 2. Monosynaptic excitatory postsynaptic potentials (EPSPs) evoked in CA1 pyramidal cells in post SSLSE tissue were always longer than those evoked in control tissue, irrespective of whether hyperresponsiveness was present or not. EPSPs elicited by stimulus subthreshold for action potentials (APs) in post-SSLSE and in control slices and matched in amplitude had a statistically greater duration in the post-SSLSE slices. Durations of monosynaptic EPSPs elicited by stimuli subthreshold for APs in DG granule cells in post-SSLSE slices were not longer than EPSPs of equal amplitude elicited in control slices. 3. Higher-intensity stimuli produced EPSPs with associated APs and, in certain cases in the post-SSLSE tissue, hyperresponsive events with multiple (> or = 3) APs. Durations of depolarizing profiles with stimuli producing APs were overall longer in both CA1 pyramidal cells and DG granule cells and correlated with the degree of hyperresponsiveness. 4. Neither the amplitudes nor the durations of monosynaptic EPSPs evoked in CA1 pyramidal cells in slices from control animals were affected by the addition of D(-)-2-amino-5-phosphonovaleric acid (APV), a blocker of the N-methyl-D-aspartate (NMDA) receptor, to the artificial cerebrospinal fluid (ACSF) bathing the slices. In contrast to the situation in control tissue, in post-SSLSE tissue APV shortened EPSPs evoked in CA1 pyramidal cells while not changing their amplitudes. After APV, inhibitory postsynaptic potentials (IPSPs) remained greatly diminished or absent in CA1 pyramidal cells. APV did not statistically decrease amplitudes of monosynaptic EPSPs evoked in DG granule cells in either control slices or post-SSLSE slices. APV decreased EPSP durations in both types of slices, more so in the post-SSLSE tissue. 5. In control slices, APV did not change the amplitudes or durations of depolarizing profiles of responses evoked by stimuli producing APs in CA1. Similarly, APV did not change the amplitudes of such responses in DG. However, APV did reduce the durations of such responses in DG in control slices.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 7472388 TI - Mechanical and electromyographic analysis of reciprocal inhibition at the human ankle joint. AB - 1. The purpose of the present study is to investigate how reciprocal inhibition influences the mechanical and electromyographic (EMG) properties of the ankle plantar flexors in humans during a voluntary contraction. 2. At different levels of maintained plantar flexion contractions ranging from 0 to 20 Nm, the size of the soleus EMG stretch reflex and the ankle joint stiffness (ration between the torque increment and the amplitude of the stretch) were measured in response to an imposed dorsiflexion. At matched plantar flexion contraction levels, stretch responses were compared before and after reversible block of the common peroneal nerve (CPN). Stretch responses were also measured during an attempted voluntary fictive dorsiflexion after CPN block. 3. In the preactivated soleus muscles, the phasic EMG response to stretch consisted of two peaks labeled M1 and M2. After CPN block, the M1 short-latency stretch reflex on average increased by 25 +/- 5.7%, mean +/- SD (P < 0.001), and the M2 stretch reflex increased on average by 29 +/- 13.0% (P = 0.002). 4. The total stiffness of the ankle joint during a stretch is the sum of the nonreflex and the reflex mediated stiffness. The total stiffness after CPN block increased on average by 13 +/- 2.7% (P = 0.002) and the estimated reflex stiffness by 33 +/- 6.5% (P < 0.001). 5. When the subjects were asked to make a strong dorsiflexion after CPN block, the soleus stretch reflex was depressed to the extent that the reflex mediated mechanical effect around the ankle joint was abolished.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7472389 TI - GABA- and glycine-activated currents in the rod bipolar cell of the rabbit retina. AB - 1. Voltage- and ligand-gated currents were recorded from solitary rabbit rod bipolar cells using the whole cell patch-clamp technique. The rod bipolar cell forms a single, stereotypical physiological and morphological class of cells that was easily identified from other neurons and support cells after enzymatic and mechanical dissociation from isolated retina. Protein kinase C immunoreactivity confirmed the validity of using a purely morphological identification of this cell type. 2. Voltage steps in 15-mV increments from a holding potential of -45 mV elicited a large outward current activated near -30 mV. These voltage-gated currents were eliminated by using equimolar substitutions of Cs+ and tetraethylammonium+ for K+ in the pipette, indicating that they represent a mixture of K+ currents. 3. The putative inhibitory neurotransmitters gamma aminobutyric acid (GABA) and glycine activated inward Cl- currents when pressure applied from pipettes placed near the axon terminals of rod bipolar cells, which were voltage-clamped at -45 mV. With changes in intracellular or extracellular Cl concentration, the reversal potential of these ligand-gated currents changed as predicted by the Nernst equation for Cl- activity. The dose-response curves for GABA and glycine were sigmoidal with saturating concentrations of 100 and 300 microM, respectively. 4. GABA-activated currents were 1) reversibly reduced by the allosteric inhibitor picrotoxin and the competitive antagonist bicuculline; 2) potentiated by the benzodiazepine diazepam and the barbiturate barbital sodium; and 3) indistinguishable from muscimol-activated currents. There was no response to the GABAB agonist baclofen. Collectively, these data strongly suggest that the GABA-activated currents in rabbit rod bipolar cells are mediated by the GABAA receptor. This is similar to the GABA-activated currents in other mammalian rod bipolar cells. 5. Application of the conformationally restricted GABA analogue cis-4-aminocrotonic acid (CACA) failed to elicit a response, whereas the conformationally extended GABA analogue trans-4-aminocrotonic acid (TACA) elicited a response similar to that of GABA. Although bicuculline appeared to suppress the GABA-activated current slightly more than the TACA-activated current (not significant using Student's t-distribution), GABA- and TACA-activated currents were equally suppressed by picrotoxin and equally enhanced by diazepam and barbital sodium. These data, coupled with the inefficacy of CACA, argue against the existence of a GABAC-type channel in the rod bipolar cell of the rabbit and suggest that GABA and TACA were activating the same GABAA receptor channel complex.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 7472390 TI - Low-voltage activated calcium currents increase in basal forebrain neurons from aged rats. AB - 1. Whole cell patch-clamp recordings were made of low-voltage-activated (LVA) calcium (Ca2+) currents using 2 mM barium (Ba2+) as charge carrier. Acutely dissociated neurons from medial septum (MS) and the nucleus of the diagonal band (nDB) were examined in young adult (1-3 mo) and aged (24-26 mo) Fischer 344 rats. 2. Most neurons in both age groups displayed LVA currents: 84% of young cells (110/131) and 87% in aged cells (62/71). Using cell capacitance as an indication of cell size, aged cells were significantly smaller (P < 0.05; 15.4 +/- 0.6 pF; mean +/- SE) than young cells (18.0 +/- 0.5 pF), although a single distribution of cell sizes was present in both populations. 3. The LVA currents were enhanced in cells from aged animals. When LVA currents were studied without activation of high voltage activated currents, the current density (pA/pF) was significantly (P < 0.05) increased at negative potentials in aged neurons (young: 4.92 +/- 0.35 pA/pF; Aged: 5.92 +/- 0.45 pA/pF, at a prepulse potential of -110 mV). No change in voltage-dependent activation or inactivation was seen. The time course of recovery from inactivation also was unchanged. 4. Kinetic parameters of LVA currents were compared in both age groups. No age-related difference in time dependent activation or inactivation was observed. A single distribution of decay time constants of LVA currents was present in both age groups. 5. These results show that MS/nDB cells maintain robust LVA currents and have increased current densities in very old rats. An increased LVA current in the aged neurons suggests that their ability to fire rhythmically or in bursts is retained or enhanced and that the resulting increase in intracellular Ca2+ may contribute to an altered Ca2+ homeostasis. PMID- 7472391 TI - Spreading depression in isolated spinal cord. AB - 1. Isolated spinal cord of amphibians is able to express Leao's spreading depression (SD). 2. SD can be evoked when the spinal cord is in an appropriated medium. There are two conditions that favor its elicitation: hypotonicity and low [Cl-] levels in the extracellular microenvironment. 3. Spinal cord SD is accompanied by a stereotyped negative extracellular voltage transient of 17.3 +/- 4.9 (standard deviation) mV amplitude and 1.2 +/- 0.5 min duration concomitantly with a reversible [K+]o increase up to 21.1 +/- 4.6 mM in the extracellular fluid. 4. Potassium salt solution can trigger SD waves that spread at 12 +/- 4.7 mm/min. 5. SD waves occur "spontaneously" in preparations superfused by Ringer solutions with very low [Cl-] (< 20 mM). The frequency of these waves decreases, and they may stop if [Mg2+] is increased. PMID- 7472392 TI - Nitric oxide is used as an orthograde cotransmitter at identified histaminergic synapses. AB - 1. Nitric oxide (NO) is produced by the enzyme nitric oxide synthase (NOS) and serves as an interneuronal messenger. Here, an identified histaminergic neuron (C2) in the mollusk Aplysia californica is shown to contain NOS with the use of NADPH diaphorase staining, suggesting that NO and histamine are used as cotransmitters by neuron C2. 2. The NOS containing neuron, C2, evokes a very slow excitatory postsynaptic potential in two of its identified postsynaptic follower neurons that are insensitive to H1 and H2 histamine receptor antagonists. 3. The very slow excitatory postsynaptic potential is blocked by inhibitors of NOS, such as nitroarginine methyl ester, and suppressed by the NO scavenger reduced hemoglobin. 4. Treatments with compounds that release NO, such as nitrosocysteine, mimic the membrane depolarization and the decrease in membrane conductance in the follower that are characteristic of the very slow excitatory postsynaptic potential induced normally by the presynaptic C2 neuron. 5. These results indicate that NO is used as an orthograde synaptic cotransmitter at synapses between histaminergic neuron C2 and its followers that receive the very slow excitatory postsynaptic potential. PMID- 7472393 TI - Role of metabotropic glutamate receptor subtypes in the patterning of epileptiform activities in vitro. AB - 1. Epileptiform activities were elicited from the in vitro guinea pig hippocampus by the addition of picrotoxin. Modification of the picrotoxin-induced activities by agents active at metabotropic glutamate receptors (mGluRs) was examined using intracellular and extracellular recordings. 2. Picrotoxin typically elicited synchronized discharges (epileptiform bursts) in CA3 neurons. These spontaneously occurred at regular intervals. In the presence of (+)-alpha-methyl-4 carboxyphenylglycine (MCPG; 700-1,000 microM), an antagonist at multiple mGluR subtypes, the frequency of spontaneous epileptiform bursts decreased. In contrast, when the mGluR agonists (1S,3R)-1-aminocyclopentane-1,3-dicarboxylic acid (ACPD; 5 microM) or (2S,3S,4S)-alpha-(carboxycyclopropyl)-glycine (L-CCG-I; 10 microM) were added to the incubating medium, the frequency of epileptiform bursts increased. No consistent change in membrane potential, burst duration, nor burst afterhyperpolarization was associated with the changes in burst frequency. 3. When spontaneous burst frequency was reduced in MCPG, stimulation at a higher frequency entrained bursts without failure. Bursts evoked in MCPG were similar in waveform and amplitude to those evoked in the control state. 4. (S)-4 carboxyphenylglycine (S-4CPG) and (R,S)-4-carboxy-3-hydroxyphenylglycine (RS 4C3HPG) are antagonists at mGluR subtypes 1 and 5 but agonists at mGluRs 2 and 3. Addition of either of these agents increased the frequency of epileptiform bursts. 5. These results suggest that sufficient glutamate is released during epileptiform activities to activate mGluRs. The overall effect is to increase the frequency of synchronized discharges. This modulatory action on burst frequency is probably mediated via the mGluR 2 and 3 receptor subclass. PMID- 7472394 TI - Motor unit activity during isometric and concentric-eccentric contractions of the human first dorsal interosseus muscle. AB - 1. Motor unit activity was recorded with intramuscular fine wire electrodes during isometric, concentric, and eccentric activity in the human first dorsal interosseus muscle. Twenty-one units from 11 subjects were sampled. 2. During isotonic cycles of shortening and lengthening, 18 of 21 units were recruited during the concentric phase, increased their discharge rates as the concentric movement progressed, then decreased their discharge rate during the eccentric phase, and were derecruited. 3. A different pattern of recruitment was observed in recordings from three units. These units were recruited during the eccentric phase, at a time when other units were decreasing their discharge rate or being derecruited. In two of the units selectively recruited during the eccentric phase, it was possible to determine their isometric thresholds, which were higher than those of units exhibiting the more common pattern of recruitment. 4. For two of the three units exhibiting selective recruitment during eccentric contraction, the unit was recorded simultaneously with different pairs of recording wires separated by 5-10 mm. Each discharge of these units was detected by both electrodes, making it unlikely that movement artifact was responsible for the initiation or cessation of discharge. 5. The recruitment patterns observed suggest that changes in the type or distribution of synaptic inputs to motoneurons during movement can, in some instances, override pre- and postsynaptic factors that shape recruitment order in isometric conditions. PMID- 7472395 TI - Postural and synergic control for three-dimensional movements of reaching and grasping. AB - 1. A fundamental question about motor control is related to the nature of the representations used by the nervous system to program the movement. Theoretically, arm displacement can be encoded either in task (extrinsic) or in joint (intrinsic) space. 2. The present study investigated the organization of complex movements consisting of reaching and grasping a cylindrical object presented along different orientations in space. In some trials, object orientation was suddenly modified at movement onset. 3. At a static level, the final limb angles were highly predictable despite the wide range of possible postures allowed by articular redundancy. Moreover, when object orientation was unexpectedly modified at movement onset, the final angular configuration of the limb was identical to that obtained when the object was initially presented along the orientation reached after the perturbation. 4. At a dynamical level, a generalized synergy was observed, and tight correlations were noted between all joint angles implicated in the movement with the exception of elbow flexion. For this joint angle, which did not vary monotonically, strong partial correlations were however observed before and after movement reversal. 5. These results suggest that natural movements are mostly carried out in joint space by postural transitions. PMID- 7472396 TI - Preferential neuronal loss in layer III of the medial entorhinal cortex in rat models of temporal lobe epilepsy. AB - We recently described a pronounced neuronal loss in layer III of the entorhinal cortex (EC) in patients with intractable temporal lobe epilepsy (Du et al., 1993a). To explore the pathophysiology underlying this distinct neuropathology, we examined the EC in three established rat models of epilepsy using Nissl staining and parvalbumin immunohistochemistry. Adult male rats were either electrically stimulated in the ventral hippocampus for 90 min or injected with kainic acid or lithium/pilocarpine. Animals were observed for behavioral changes for up to 6 hr and were killed 24 hr or 4 weeks after the experimental treatments. At 24 hr, all animals that had exhibited a bout of acute status epilepticus showed a consistent pattern of neuronal loss in the EC in Nissl stained sections. Neurodegeneration was most pronounced in layer III of the medial Ec at all dorsoventral levels. A few surviving neurons were frequently present in the lesioned area. An identical pattern of nerve cell loss was also seen in the EC of rats killed 4 weeks following the treatments. This lesion was completely prevented by an injection of diazepam and pentobarbital, given 1 hr after kainic acid administration. Immunohistochemistry demonstrated a relative resistance of parvalbumin-positive neurons in layer III of the medial EC. Taken together, these experiments indicate that prolonged seizures cause a preferential neuronal loss in layer III of the medial EC and that this lesion may be related to a pathological elevation of intracellular calcium ion concentrations. PMID- 7472397 TI - Distribution of an APP homolog, APLP2, in the mouse olfactory system: a potential role for APLP2 in axogenesis. AB - Deposition of beta-amyloid (A beta) in senile plaques is a major pathological characteristic of Alzheimer's disease. A beta is generated by proteolytic processing of amyloid precursor proteins (APP). APP is a member of a family of related polypeptides that includes amyloid precursor-like proteins APLP1 and APLP2. To examine the distribution of APLP2 in the nervous system, we generated antibodies specific for APLP2 and used these reagents in immunocytochemical and biochemical studies of the rodent nervous system. In this report, we document that in cortex and hippocampus, APLP2 is enriched in postsynaptic compartments. In the olfactory system, however, APLP2 is abundant in olfactory sensory axons, and axon terminals in glomeruli. Confocal microscopy revealed that APLP2 is present in both pre- and postsynaptic compartments in the olfactory bulb. Notably, mRNA encoding chondroitin sulfate glycosaminoglycan (CS GAG)-modified forms of APLP2 are enriched in the olfactory epithelium, relative to alternatively-spliced mRNA, encoding CS GAG-free forms of APLP2. In addition, we demonstrate that CS-modified APLP2 forms accumulate in the olfactory bulb. CS proteoglycans are known to play an important role in regulating cell migration and neuronal outgrowth. Since sensory neurons in the olfactory epithelium are in a state of continual turnover, axons of newly generated cells must establish synaptic connections with neurons in the olfactory bulb in adult life. The presence of APLP2 in olfactory sensory axons and glomeruli is consistent with the view that this protein may play an important role in axonal pathfinding and/or synaptogenesis. PMID- 7472398 TI - Persistent polyneuronal innervation in partially denervated rat muscle after reinnervation and recovery from prolonged nerve conduction block. AB - The contribution of activity to the long-term stability of synaptic connections is a subject of ongoing debate. In the present study we examined the effects of recovery from chronic disuse on the pattern of reinnervation of partially denervated adult rat skeletal muscles, using tension measurements, intracellular recordings, and observations of vital staining with activity-dependent styryl dyes. Fourth deep lumbrical muscles were partially denervated by crushing the lateral plantar nerve (LPN) bilaterally. Denervated muscle fibers became innervated by sprouts from the sural nerve (SN); 17-23 d after LPN crush, conduction in the right sciatic nerve was blocked by connecting an osmotic minipump containing tetrodotoxin to a cuff placed around the nerve. Distal muscles remained continuously paralysed for 10-19 d. After 2 weeks of nerve block the tension produced by stimulating the regenerated LPN axons had reached about 70% of the total. Regenerating axons in contralateral muscles reinnervated only about 55% of the muscle fibers. The level of dual innervation by both regenerating and intact axons reached about 50% of the total muscle fibers after 2 weeks of paralysis, but only about 20% in contralateral controls. We then measured the rate and amount of decline in motor unit tension and polyneuronal innervation in reinnervated muscles during an eight week period of recovery from nerve block. Some LPN and SN connections regressed within 2-4 weeks, but about 35% of the muscle fibers still retained convergent inputs from LPN and SN motor axons 8 weeks after activity had resumed. This was about twice the level observed in contralateral reinnervated muscles. Vital staining with the styryl dyes FM1-43 and RH414 confirmed that many of the reinnervated motor end-plates were convergently supplied by both SN and LPN axons. Intracellular recordings showed that most of the dually innervated fibers in paralyzed muscles were supplied by suprathreshold inputs from both LPN and SN axons. The increased excitability of these muscle fibers was partly explained by their two-fold increased input resistance. Input resistance recovered to control levels within 4 weeks of resumption of activity, but dually innervated fibers in previously blocked muscles still gave mostly suprathreshold responses to stimulation of both the LPN and the SN. We conclude that chronic nerve conduction block generates or sustains a local environment which allows some convergent synaptic inputs on reinnervated muscle fibers to become consolidated and strengthened, independent of subsequent neuromuscular activity. PMID- 7472399 TI - Localization of novel corticotropin-releasing factor receptor (CRF2) mRNA expression to specific subcortical nuclei in rat brain: comparison with CRF1 receptor mRNA expression. AB - Corticotropin-releasing factor (CRF) is the primary factor involved in controlling the release of ACTH from the anterior pituitary and also acts as a neurotransmitter in a variety of brain systems. The actions of CRF are mediated by G-protein coupled membrane bound receptors and a high affinity CRF receptor, CRF1, has been previously cloned and functionally characterized. We have recently isolated a cDNA encoding a second member of the CRF receptor family, designated CRF2, which displays approximately 70% homology at the nucleotide level to the CRF1 receptor and exhibits a distinctive pharmacological profile. The present study utilized in situ hybridization histochemistry to localize the distribution of CRF2 receptor mRNA in rat brain and pituitary gland and compared this with the distribution of CRF1, receptor expression. While CRF1 receptor expression was very high in neocortical, cerebellar, and sensory relay structures, CRF2 receptor expression was generally confined to subcortical structures. The highest levels of CRF2 receptor mRNA in brain were evident within the lateral septal nucleus, the ventromedial hypothalamic nucleus and the choroid plexus. Moderate levels of CRF2 receptor expression were evident in the olfactory bulb, amygdaloid nuclei, the paraventricular and suraoptic nuclei of the hypothalamus, the inferior colliculus and 5-HT-associated raphe nuclei of the midbrain. CRF2-expressing cells were also evident in the bed nucleus of the stria terminalis, the hippocampal formation and anterior and lateral hypothalmic areas. In addition, CRF2 receptor mRNA was also found in cerebral arterioles throughout the brain. Within the pituitary gland, CRF2 receptor mRNA was detectable only at very low levels in scattered cells while CRF1 receptor mRNA was readily detectable in anterior and intermediate lobes. This heterogeneous distribution of CRF1 and CRF2 receptor mRNA suggests distinctive functional roles for each receptor in CRF related systems. The CRF1 receptor may be regarded as the primary neuroendocrine pituitary CRF receptor and important in cortical, cerebellar and sensory roles of CRF. The anatomical distribution of CRF2 receptor mRNA indicates a role for this novel receptor in hypothalamic neuroendocrine, autonomic and general behavioral actions of central CRF. PMID- 7472400 TI - Cerebellar precursors transplanted to the neonatal dentate gyrus express features characteristic of hippocampal neurons. AB - During the development of the CNS, a salient issue is whether neuronal phenotype is defined by the lineage or by the environment of precursor cells. Transplants permit these two possibilities to be tested, as cell fate can be examined in a new location. Dissociated cerebellar cells from newborn rats treated with tritiated thymidine or from NSE-lacZ transgenic mice were grafted into the dentate gyrus of the developing hippocampus. Implanted cells integrated into the granule cell layer, which contains the cell bodies of host granule neurons. Immunohistochemistry showed that grafted cells in the granule cell layer, like the host hippocampal granule neurons, were calbindin positive and upregulated FOS in a seizure paradigm. Electron microscopic analysis also showed that cells grafted to the dentate gyrus share features with host dentate neurons. These assays indicate that transplanted cerebellar cells acquired morphological and antigenic features characteristic of hippocampal neurons. These results show that metencephalic precursors are capable of differentiating in response to signals in the telencephalon, suggesting that the environment controls the regional fate of neuronal precursor cells during neurogenesis. PMID- 7472401 TI - Upregulation of bax protein levels in neurons following cerebral ischemia. AB - The patterns of expression of the bcl-2, bax, and bci-X genes were examined immunohistochemically in neurons of the adult rat brain before and after 10 min of global ischemia induced by transient cardiac arrest. High levels of the cell death promoting protein Bax and concomitant low levels of the apoptosis-blocking protein Bcl-2 were found in some populations of neurons that are particularly sensitive to cell death induced by transient global ischemia, such as the CA1 sector of the hippocampus and the Purkinje cells of the cerebellum. Moreover, within 0.5 to 3 hr after an ischemic episode, immunostaining for Bax was markedly increased within neurons with morphological features of degeneration in many regions of the brain. Use of a two-color staining method for simultaneous analysis of Bax protein and in situ detection of DNA-strand breaks revealed high levels of Bax immunoreactivity in many neurons undergoing apoptosis. Postischemic elevations in Bax protein levels in the hippocampus, cortex, and cerebellum were also demonstrated by immunoblotting. At early times after transient ischemia, regulation of Bcl-2 and Bcl-x protein levels varied among neuronal subpopulations, but from 3 hr on, those neurons with morphological evidence of degeneration uniformly contained reduced levels of Bci-2 and particularly Bci-X immunoreactivity. The findings suggest that differential expression of some members of the bcl-2 gene family may play an important role in determining the relative sensitivity of neuronal subpopulations to ischemia and that postischemic alterations in the expression of bax, bcl-2, and bcl-x may contribute to the delayed neuronal cell death that occurs during the repurfusion phase after a transient ischemic episode. PMID- 7472402 TI - Mitochondrial production of reactive oxygen species in cortical neurons following exposure to N-methyl-D-aspartate. AB - Increasing evidence suggests that glutamate neurotoxicity is partly mediated by reactive oxygen species, formed as a consequence of several processes, including arachidonic acid metabolism and nitric oxide production. Here we used an oxidation-sensitive indicator, dihydrorhodamine 123, in combination with confocal microscopy, to examine the hypothesis that electron transport by neuronal mitochondria may be an important source of glutamate-induced reactive oxygen species (ROS). Exposure to NMDA, but not kainate, ionomycin, or elevated potassium stimulated oxygen radical production in cultured murine cortical neurons, demonstrated by oxidation of nonfluorescent dihydrorhodamine 123 to fluorescent rhodamine 123. Electron paramagnetic resonance spectroscopy studies using 5,5-dimethyl-1-pyrroline-N-oxide (DMPO) as a radical-trapping agent, also showed production of ROS by cortical neurons after NMDA but not kainate exposure. NMDA-induced ROS production depended on extracellular Ca2+, and was not affected by inhibitors of nitric oxide synthase or arachidonic acid metabolism. The increased production of ROS was blocked by inhibitors of mitochondrial electron transport, rotenone or antimycin, and mimicked by the electron transport uncoupler, carbonyl cyanide p-trifluoromethoxyphenylhydrazone. These data support the possibility that NMDA receptor-mediated, Ca(2+)-dependent uncoupling of neuronal mitochondrial electron transport may contribute to the oxidative stress initiated by glutamate exposure. PMID- 7472403 TI - The first retinal axon growth in the mouse optic chiasm: axon patterning and the cellular environment. AB - The retinofugal pathway is a useful model for axon guidance because fibers from each eye project to targets on both sides of the brain. Studies using static and real time analyses in mice at E15-17 demonstrated that uncrossed axons from ventrotemporal retina diverge from crossed axons in the optic chiasm, where specialized resident cells may direct divergence. Other studies, however, suggest that pioneering uncrossed retinal axons derive from a different retinal region, take a different course, and enter the ipsilateral optic tract independent of fiber-fiber interactions. We examine these differences by dye-labeling the earliest optic axons and immunocytochemically identifying cells in their path. The first optic axons arising from dorsocentral retina, enter the diencephalon at E12.5. All axons initially grow caudally, lateral to a radial glial palisade. In contrast to later growing axons, early uncrossed axons enter the ipsilateral optic tract directly. Crossed axons enter the glial palisade and course medially, then anteriorly, in a pathway corresponding to the border of an early neuronal population that expresses SSEA-1, CD44, and beta-tubulin. Axon patterning occurs independent of fiber-fiber interactions from both eyes, as the first uncrossed axons enter the optic tract before crossed ones from opposite eye. These analysis, in conjunction with our previous studies during the principal period of retinal axon growth in the diencephalon, suggest that the adult visual projection arises from age-dependent variations in the types and relative contribution of cues along the path through the emerging optic chiasm. PMID- 7472404 TI - Immunochemical identification and subcellular distribution of the alpha 1A subunits of brain calcium channels. AB - A site-directed anti-peptide antibody (anti-CNA1) directed against the alpha 1 subunit of class A calcium channels (alpha 1A) recognized a protein of approximately 190-200 kDa in immunoblot and immunoprecipitation analyses of rat brain glycoproteins. Calcium channels recognized by anti-CNA1 were distributed throughout the brain with a high concentration in the cerebellum. Calcium channels having alpha 1A subunits were concentrated in presynaptic terminals making synapses on cell bodies and on dendritic shafts and spines of many classes of neurons and were especially prominent in the synapses of the parallel fibers of cerebellar granule cells on Purkinje neurons where their localization in presynaptic terminals was confirmed by double labeling with the synaptic membrane protein syntaxin or the microinjected postsynaptic marker Neurobiotin. They were present in lower density in the surface membrane of dendrites of most major classes of neurons. There was substantial labeling of Purkinje cell bodies, but less intense staining of the cell bodies of hippocampal pyramidal neurons, layer V pyramidal neurons in the dorsal cortex, and most other classes of neurons in the forebrain and cerebellum. Scattered cell bodies elsewhere in the brain were labeled at low levels. These results define a unique pattern of localization of class A calcium channels in the cell bodies, dendrites, and presynaptic terminals of most central neurons. Compared to class B N-type calcium channels, class A calcium channels are concentrated in a larger number of presynaptic nerve terminals implying a more prominent role in neurotransmitter release at many central synapses. PMID- 7472405 TI - Biochemical properties and subcellular distribution of the neuronal class E calcium channel alpha 1 subunit. AB - Anti-peptide antibodies specific for the neuronal calcium channel alpha 1E subunit (anti-CNE1 and anti-CNE2) were produced to study the biochemical properties and subcellular distribution of the alpha 1E polypeptide from rat brain. Immunoblotting identified a single size form of 245-255 kDa which was a substrate for phosphorylation by cAMP-dependent protein kinase, protein kinase C, cGMP-dependent protein kinase, and calcium/calmodulin-dependent protein kinase II. Ligand-binding studies of alpha 1E indicate that it is not a high affinity receptor for the dihydropyridine isradipine or the peptide toxins omega-conotoxin GVIA or omega-conotoxin MVIIC at concentrations which elicit high affinity binding to other channel types in the same membrane preparation. The alpha 1E subunit is widely distributed in the brain with the most prominent immunocytochemical staining in deep midline structures such as caudate-putamen, thalamus, hypothalamus, amygdala, cerebellum, and a variety of nuclei in the ventral midbrain and brainstem. Staining is primarily in the cell soma but is also prominent in the dendritic field of a discrete subset of neurons including the mitral cells of the olfactory bulb and the distal dendritic branches of the cerebellar Purkinje cells. Our observations indicate that the 245-255 kDa alpha 1E subunit is localized in cell bodies, and in some cases in dendrites, of a broad range of central neurons and is potentially modulated by multiple second messenger-activated protein kinase. PMID- 7472406 TI - Role of calcium channel subtypes in calcium transients in hippocampal CA3 neurons. AB - Multiple subtypes of voltage-gated calcium channels are differentially localized in brain neurons suggesting that they serve distinct roles in neuronal excitation and signaling. In organotypic hippocampal slice cultures, class D (L-type) calcium channels are predominantly located in the cell bodies of CA3 neurons while class B (N-type) and class A (P or Q-type) are localized in dendrites and associated presynaptic terminals with relatively low somal expression. Using specific antagonists to inhibit calcium transients recorded in CA3 neuronal cell bodies, we found that L-type calcium channels have a predominant role in somal calcium transients elicited by trains of strong stimuli applied to either the soma or the distal apical dendrite while class A calcium channels make a smaller contribution. Presynaptic class B (N-type) and class A (P- and/or Q-type) calcium channels are critical for glutamate-mediated synaptic transmission onto the dendrites of CA3 neurons. Postsynaptic class A and B calcium channels detected on the dendritic shaft by immunocytochemistry were not found to contribute substantially to somal calcium transients during repetitive stimulation of distal dendrites, but sodium channels were required for calcium transients elicited by somatic or dendritic stimulation. Our results show that the different calcium channel subtypes serve distinct roles in cellular activation and transmission of signals in CA3 neurons, consistent with their differential subcellular localization. PMID- 7472407 TI - Axotomy-induced axonal degeneration is mediated by calcium influx through ion specific channels. AB - We examined the role of extracellular calcium entry, the possible involvement of axonal calcium channels, and the potential protective effect of calcium channel and calpain antagonists in axotomy-induced axonal degeneration using murine dorsal root ganglia in cell culture. We found that calcium entry is both necessary and sufficient to induce axonal degeneration after axotomy, and may be inhibited by cobalt, manganese, dihydropyridines, and bepridil. Tetrodotoxin and omega-conotoxin are ineffective in preventing axonal degeneration. The activation of calpains also appears to be necessary and sufficient for axonal degeneration to proceed, and can be blocked with membrane-permeant leupeptin analogs and the oxirane aloxistatin. Although other calcium-activated events may occur, it appears that inhibition of calpain is sufficient to preserve the axon at the light microscope level, and to prevent axonal cytoskeleton degradation as detected by immunofluorescent staining. Our results suggest that axonal degeneration after axotomy involves the following sequence of events: (1) a lag period after axotomy prior to the onset of axonal degeneration, (2) entry of calcium into the axon through an intact axolemma via a calcium-specific ion transport mechanism, (3) activation of calcium-dependent effector molecules such as calpains, (4) degradation of the axonal cytoskeleton. The details of the second step require further elucidation, and are of particular interest because this step is a potential target for therapies directed towards peripheral neuropathies. PMID- 7472408 TI - The nicotinic blocking agents d-tubocurare and alpha-bungarotoxin save motoneurons from naturally occurring death in the absence of neuromuscular blockade. AB - Motoneurons undergo a phase of target-dependent cell death during development. In chick embryos, motoneuronal death is blocked by the application of the nicotinic antagonists d-tubocurare (dTC) or alpha-bungarotoxin (alpha BTX). Paralytic doses of these drugs also increase intramuscular nerve branch formation. To investigate the possibility that a neuronal rather than a muscle-type nicotinic ACh receptor (nAChR) might be responsible for the toxin-induced arrest of naturally occurring motoneuronal death, we compared the doses of dTC and alpha-BTX required for paralysis with those needed to protect motoneurons from cell death. We also measured the effects of dTC on the survival of retrogradely labeled motoneurons in culture, and of various doses of dTC on intramuscular nerve branch formation. Subparalytic doses of dTC caused small but significant increases in nerve branch number, while higher doses produced larger effects. In contrast, motoneuronal survival was already maximal at doses of dTC or alpha-BTX below those needed for a visible effect on limb movement. Moreover, dTC increased motoneuron survival in culture, in the absence of muscle cells and muscle-type nAChRs. Nicotinic blocking agents can therefore rescue motoneurons with minimal depression of neuromuscular transmission, suggesting that this effect may be mediated through neuronal, rather than muscle-type. PMID- 7472409 TI - Transfer of coded information from sensory to motor networks. AB - During sensory-guided motor tasks, information must be transferred from arrays of neurons coding target location to motor networks that generate and control movement. We address two basic questions about this information transfer. First, what mechanisms assure that the different neural representations align properly so that activity in the sensory network representing target location evokes a motor response generating accurate movement toward the target? Coordinate transformations may be needed to put the sensory data into a form appropriate for use by the motor system. For example, in visually guided reaching the location of a target relative to the body is determined by a combination of the position of its image on the retina and the direction of gaze. What assures that the motor network responds to the appropriate combination of sensory inputs corresponding to target position in body- or arm-centered coordinates? To answer these questions, we model a sensory network coding target position and use it to drive a similarly modeled motor network. To determine the actual motor response we use decoding methods that have been developed and verified in experimental work. We derive a general set of conditions on the sensory-to-motor synaptic connections that assure a properly aligned and transformed response. The accuracy of the response for different numbers of coding cells is computed. We show that development of the synaptic weights needed to generate the correct motor response can occur spontaneously through the observation of random movements and correlation-based synaptic modification. No error signal or external teaching is needed during this process. We also discuss nonlinear coordinate transformations and the presence of both shifting and nonshifting receptive fields in sensory/motor systems. PMID- 7472410 TI - Phosducin and PP33 are in vivo targets of PKA and type 1 or 2A phosphatases, regulators of cell elongation in teleost rod inner-outer segments. AB - Teleost rod photoreceptors elongate in the light and shorten in darkness. We are investigating the role of cAMP-dependent protein kinase (PKA), phosphatases and target phosphoproteins in the regulation of photoreceptor cell shape. Preparations of rod fragments, consisting of the motile inner segment with attached photosensory outer segment (RIS-ROS), undergo light-stimulated elongation in culture. The PKA-selective inhibitor, H89, enhanced RIS-ROS elongation in both light and darkness, suggesting that elongation is associated with dephosphorylation of PKA substrates. Okadaic acid and calyculin A, inhibitors of type 1 and 2A phosphatases, blocked light-dependent and light independent elongation with relative potencies suggesting that elongation requires dephosphorylation by type 1 phosphatase in light and type 2A phosphatase in darkness. To identify targets of PKA and phosphatases, RIS-ROS were isolated from retinas prelabeled with 32P-orthophosphate, and then incubated in the presence of kinase inhibitors or phosphatase inhibitors. Two phosphoproteins, PP33 and PP35, were phosphorylated by PKA and dephosphorylated by type 1 or 2A phosphatases in light- and dark-cultured RIS-ROS. PP35 (but not PP33) was immunoprecipitated by an antibody to phosducin, a PKA-regulated modulator of phototransduction (Lee et al., 1992); PP35 was also phosphorylated in vitro by a Ca2+ calmodulin-activated kinase. PP33 further differed from PP35 in its phosphopeptide maps and phosphorylation by PKC. We conclude that RIS-ROS elongation is correlated with the dephosphorylation of PKA substrates by type 1 or 2A phosphatases. Candidate mediator proteins include PP35, a fish phosducin homolog, and PP33, a newly described photoreceptor phosphoprotein. PMID- 7472411 TI - Light removes inactivation of the A-type potassium channels in scallop hyperpolarizing photoreceptors. AB - The properties of light-activated and voltage dependent conductances in the hyperpolarizing photoreceptors of the isolated retina of the giant scallop (Patinopecten yessoensis) have been studied in whole-cell voltage-clamp recordings. Resting membrane potential of the cells was -35 to -60 mV in the dark. Large outward current was maintained during illumination when the cell was voltage clamped at the resting potential. Previous reports have shown that the hyperpolarizing response to light and the light-activated outward current are mediated by an increase of a conductance of K+ channels in the plasma membrane (Gorman and McReynolds, 1974; Gomez and Nasi, 1994a). This report shows that the light-activated K+ channels of this photoreceptor cell generate the voltage dependent transient outward current in response to depolarizing voltage steps in the dark. The characteristic of this current resembles the typical voltage dependent transient current (A-current) of molluscan neurons in the kinetics of activation and inactivation, and in the blockage by 4-aminopyridine (4-AP). However, the voltage-dependent transient current during illumination becomes a flat stable form which inactivation process disappeared. We report here that light removes inactivation of the K+ channels which generate the voltage dependent transient outward currents in the dark. PMID- 7472412 TI - Cloning and characterization of chi-1: a developmentally regulated member of a novel class of the ionotropic glutamate receptor family. AB - Ionotropic glutamate receptors are composed of homomeric or heteromeric configurations of glutamate receptor subunits. We have cloned a member of a novel class of the rat ionotropic glutamate receptor family, termed chi-1. This subunit exhibits an average identity of 27% to NMDA subunits and 23% to non-NMDA subunits. Regional transcript levels of chi-1 are elevated just prior to and during the first postnatal week, with the highest levels present in the spinal cord, brainstem, hypothalamus, thalamus, CA1 field of the hippocampus, and amygdala. The spatial distribution of chi-1 expression is similar from postnatal day 1 (P1) to adulthood. However, transcript levels decline sharply between P7 and P14 and remain attenuated into adulthood. Functional expression studies in Xenopus oocytes injected with in vitro transcribed chi-1 RNA did not demonstrate agonist-activated currents. Pairwise expression of chi-1 with members of the AMPA, KA, or delta class of glutamate recepto subunits either failed to generate agonist-activated currents or failed to alter the underlying current generated by the coexpressed subunit. However, coexpression of chi-1 with subunits forming otherwise functional NMDA receptors resulted in an inhibition of current responses. Since chi-1 did not alter the currents generated by non-NMDA subunits, this suggests that chi-1 may specifically interact with NMDA receptor subunits. Further characterization will be required to establish the precise role of this glutamate receptor subunit in neuronal signaling. PMID- 7472413 TI - Developmental and regional expression pattern of a novel NMDA receptor-like subunit (NMDAR-L) in the rodent brain. AB - A novel NMDA receptor-like (NMDAR-L) cDNA was isolated that contained an open reading frame coding for a predicted polypeptide of 1115 amino acids that shares approximately 27% identity with NMDA receptor subunits. In situ hybridization experiments indicated that NMDAR-L mRNA was expressed in the developing rodent CNS. On postnatal day 1 (P1), NMDAR-L mRNA expression was pronounced in the entorhinal cortex, the subiculum and the thalamus, in layer V of the developing neocortex, in the superior and inferior colliculi, and various regions of the hindbrain, excluding the cerebellum. On P5, NMDAR-L mRNA was expressed in layer V of the neocortex, in the entorhinal cortex, in the subiculum, and in the thalamus. On P14, NMDAR-L mRNA was expressed in layers II-VI of the neocortex, in the entorhinal and piriform cortex, in the subiculum and CA1 field, and in the nucleus of the lateral olfactory tract. In the adult brain, NMDAR-L mRNA was detected predominately in the nucleus of the lateral olfactory tract. Injection of NMDAR-L cRNA into Xenopus oocytes did not lead to the expression of homomeric glutamate-activated channels. However, coinjection of the triple combination of NMDAR-L with NMDAR1 and NMDAR2B cRNAs led to a striking decrease in the current magnitude compared to currents obtained after coexpression of the double combination of NMDAR1 with NMDAR2B. While the function of NMDAR-L remains to be established, its developmental and regional expression pattern suggests that NMDAR-L may influence axonal outgrowth and synaptogenesis during brain development. PMID- 7472414 TI - Sympathetic neurons in neonatal rats require endogenous neurotrophin-3 for survival. AB - Gene deletion of neurotrophin-3 (NT3) results in severe sensory and sympathetic deficits that are incompatible with postnatal life in mice. We have now addressed the question of whether NT3 plays a role in the postnatal animal. An antiserum specific for NT3 and capable of blocking the survival effect of the factor in vitro has been generated and given to neonatal rats. Antiserum administration during either or both of the first 2 postnatal weeks resulted in a 54-74% reduction in the size of the superior cervical ganglia, reflecting a loss of as many as 80% of all neurons, with a predominant effect on the neuropeptide Y containing subpopulation. The immunoreactivities of NPY, tyrosine hydroxylase, and p75 low affinity NGF receptor in nerve terminals within the mesenteric artery were also reduced, whereas that of the sensory neuron neuropeptide, calcitonin gene related peptide was less affected. These results demonstrate that the majority of sympathetic neurons of the neonatal rat are dependent on endogenous NT3 for their survival at a time when they are also dependent on another survival factor, NGF, thus apparently providing a clear example of a population of neurons requiring for their survival the simultaneous supply of more than one trophic factor. PMID- 7472415 TI - In a rat model of parkinsonism, lesions of the subthalamic nucleus reverse increases of reaction time but induce a dramatic premature responding deficit. AB - Lesions of the subthalamic nucleus (STN) have been found to reduce the severe akinetic motor symptom produced in animal models of Parkinson's disease, such as in monkeys treated with 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP) or in monoamine-depleted rats. However, little is known about the effect of STN exclusion on subtle motor deficits induced by moderate dopaminergic lesions in complex motor tasks. The present study was thus performed on rats trained in a reaction time (RT) task known to be extremely sensitive to variations of dopamine transmission in the striatum. Animals were trained to release a lever after the onset of a visual stimulus within a time limit to obtain a food reward. Discrete dopamine depletion produced by infusing the neurotoxin 6-hydroxydopamine (6-OHDA) bilaterally into the dorsal part of the striatum, produced motor initiation deficits which were revealed by an increase in the number of delayed responses (lever release after the time limit) and a lengthening of RTs. In contrast, bilateral excitotoxic lesion of the STN with ibotenic acid induced severe behavioral deficits which were opposite to those produced by the dopaminergic lesion, as shown by an increase in the number of premature responses (lever release before the onset of the visual stimulus) and a decrease of RTs. Surprisingly, the performance of the animals bearing a double lesion (striatal dopaminergic lesion followed 14 d later by STN ibotenic lesion) was still impaired 40 d after the ibotenic lesion. As expected, the 6-OHDA-induced motor initiation deficits were reversed by a subsequent STN lesion. However, the dramatic increase of premature responses contributing to major behavioral deficits induced by the STN lesion remained unchanged. Thus, the bilateral lesion of the STN was found to alleviate the motor deficits in this model of parkinsonism, but essentially produced over time, long lasting deficits that might be related to dyskinesia or cognitive impairment. The present results strongly support the recent concept of a predominant control of the STN on basal ganglia output structures. PMID- 7472416 TI - Expression of EPSP/spike potentiation following low frequency and tetanic stimulation in the CA1 area of the rat hippocampus. AB - Low frequency stimulation (LFS, 1 Hz for 15 min) has been shown to produce an NMDA receptor dependent homosynaptic long-term depression (LTD) of synaptic potentials in the CA1 area of the rat hippocampus. Here we describe experiments aimed at characterizing EPSP/spike (E/S) coupling associated with this form of LTD. Our data show that following LFS neurons have a higher probability of synchronous discharge in response to a population EPSP of fixed slope (E/S potentiation). This E/S potentiation was not significantly enhanced by a tetanic stimulation. When the protocol was reversed, that is, starting with a tetanic stimulus, E/S potentiation was observed which was unaffected by a subsequent LFS. Saturating these synaptic responses to either a maximal or a minimal value produced similar effects on E/S coupling. E/S depression was never encountered. Finally, we found that the expression of E/S potentiation did not require the activation of NMDA receptors. These data suggest that at the level of a local neuronal network in the CA1 area, LFS is not a very powerful tool since the synaptic depression is associated with a potentiation of the population response of these neurons. Furthermore, the expression of E/S dissociation seems different from that of homosynaptic long-term potentiation and LTD. PMID- 7472417 TI - Cannabinoids activate an inwardly rectifying potassium conductance and inhibit Q type calcium currents in AtT20 cells transfected with rat brain cannabinoid receptor. AB - Rat brain cannabinoid receptor (CB-1) was stably transfected into the murine tumor line AtT-20 to study its coupling to inwardly rectifying potassium currents (Kir) and high voltage-activated calcium currents (ICa). In cells expressing CB-1 ("A-2" cells), cannabinoid agonist potently and stereospecifically activated Kir via a pertussis toxin-sensitive G protein. ICa in A-2 cells was sensitive to dihydropyridines and omega CTX MVIIC, less so to omega CgTX GVIA and insensitive to omega Aga IVa. In CB-1 expressing cells, cannabinoid agonist inhibited only the omega CTX MVIIC-sensitive component of ICa. Inhibition of Q-type ICa was voltage dependent and PTX sensitive, thus similar in character to the well studied modulation of N-type ICa. An endogenous cannabinoid, anandamide, activated Kir and inhibited ICa as efficaciously as potent cannabinoid agonist. Immunocytochemical studies with antibodies specific for class A, B, C, D, and E voltage-dependent calcium channel alpha 1 subunits revealed that AtT-20 cells express each of these major classes of alpha 1 subunit. PMID- 7472419 TI - Pharmacological characterization of presynaptic calcium currents underlying glutamatergic transmission in the avian auditory brainstem. AB - We used whole-cell patch-clamp techniques on presynaptic terminals and postsynaptic neurons of the glutamatergic magnocellularis synapse in the chick auditory brainstem to study the effects of N, P, and L-type calcium channel blockers directly on presynaptic calcium currents and transmitter release. Presynaptic calcium currents and transmitter release were unaffected by nifedipine, blocked partially by omega-agatoxin IVA and completely by omega conotoxin GVIA. The presynaptic calcium current is a low voltage-activated, noninactivating current and its block by omega-agatoxin IVA was not reversed by repeated depolarization of the presynaptic terminal. Thus, the presynaptic calcium current that underlies transmitter release at the chick magnocellularis synapse differs in some respects from N and P-type calcium currents described in vertebrate neuronal cell bodies. PMID- 7472418 TI - Subthalamic nucleus lesions: widespread effects on changes in gene expression induced by nigrostriatal dopamine depletion in rats. AB - Lesions of the subthalamic nucleus block behavioral effects of nigrostriatal dopamine depletion in rats and primates, but the contribution of this region to the molecular effects of dopaminergic lesions is unknown. The effects of subthalamic nucleus lesions alone or in combination with a 6-hydroxydopamine induced lesion of the substantia nigra were examined in adult rats. Unilateral subthalamic nucleus lesions caused ipsiversive rotation after peripheral administration of apomorphine and a small decrease in glutamic acid decarboxylase (GAD) mRNA in the ipsilateral globus pallidus (external pallidum). Confirming previous results, nigrostriatal dopaminergic lesions caused contraversive rotation after apomorphine injection, and increased enkephalin mRNA in the striatum, GAD mRNA in the globus pallidus, and somatostatin mRNA in the entopeduncular nucleus (internal pallidum) ipsilateral to the lesion. In addition, the lesion decreased substance P mRNA in the ipsilateral striatum compared to the contralateral side, and GAD mRNA in the contralateral entopeduncular nucleus. These effects were abolished in rats with lesions of the subthalamic nucleus and substantia nigra on the same side. Thus, the subthalamic lesions prevented changes in gene expression induced by dopamine depletion, not only in regions receiving a direct input from the subthalamic nucleus (ipsilateral pallidum), but also in regions which do not (striatum and contralateral pallidum). This suggests that polysynaptic pathways regulated by the subthalamic nucleus contribute to the effects of dopaminergic lesions in many regions of the basal ganglia. This pivotal role of the subthalamic nucleus may account for the beneficial effects of subthalamic nucleus lesions on motor symptoms resulting from dopamine depletion. PMID- 7472420 TI - Inhibitors of cyclic-GMP phosphodiesterase alter excitation of Limulus ventral photoreceptors in Ca(2+)-dependent fashion. AB - We have examined the hypothesis that Ca(2+)-dependent cyclic-GMP metabolism may play a role in visual transduction in Limulus photoreceptors. Although phosphoinositide hydrolysis is central to phototransduction and phosphoinositide dependent Ca(2+)-mobilization seems to be required for transduction, the subsequent steps leading to ion channel gating (the immediate cause of excitation) are not understood. Channels normally opened in response to light can be opened in excised membrane patches by cGMP but not by Ca2+, suggesting that cGMP acts as a channel ligand in excitation. Using phosphodiesterase inhibitors, we investigated whether changes in cGMP metabolism could affect excitation. We report that zaprinast and IBMX increased the amplitudes and retarded the kinetics of physiological light responses. These effects were maximal for brightest stimuli. The effects were markedly enhanced in low Ca2+ conditions. In contrast, excitation induced by direct IP3-injection and by direct Ca(2+)-injection were inhibited. These observations suggest that PI-induced excitation is dependent on cGMP metabolism in a Ca(2+)-dependent manner, and they support the possibility that transduction involves modification of cGMP metabolism by Ca(2+)-release resulting from phosphoinositide hydrolysis. PMID- 7472421 TI - Cholinergic modulation of activity-dependent synaptic plasticity in the piriform cortex and associative memory function in a network biophysical simulation. AB - The effect of activation of cholinergic receptors on long-term potentiation (LTP) in rat piriform cortex pyramidal cells was studied using extracellular and intracellular recordings in brain slice preparations. The functional role of this modulation was studied in a realistic network biophysical stimulation. Repetitive stimuli were applied in two paradigms: one in which the recorded cell was held at its resting potential and one in which synaptic activity was superimposed on a depolarizing pulse strong enough to evoke four action potentials. In the absence of cholinergic modulation, stimulation at 5 Hz induced LTP primarily in the second condition (13.7%, n = 6 out of 9, measured at 10 min after tetanus). When stimuli were applied in the presence of the muscarinic agonist carbachol (20 microM), LTP of greater amplitude was induced in both paradigms (resting: 41.5%, n = 11 out of 16, depolarized: 36%, n = 5 out of 7, measured at 10 min after tetanus). Increases in excitatory postsynaptic potential (EPSP) amplitudes in the presence of carbachol were gradual, starting at the time 5 Hz stimuli were applied and continuing until an action potential was evoked synaptically. In the presence of the NMDA receptor antagonist 2-amino-5-phosphonovaleric acid (APV), LTP could not be induced. The muscarinic antagonist atropine also prevented LTP induction in the presence of carbachol. Cholinergic modulation of synaptic plasticity was examined in a previously developed realistic biophysical network simulation. In simulations, use of a gradual rate of synaptic modification prevented excessive strengthening of synapses, which could cause interference between stored patterns. The effect of excess synaptic strengthening can be avoided by introducing activity dependent depression of synaptic strength. Coactivation of learning and depression rules results in a stable system where no interference occurs, at any rate of learning. Implementing the depression rule only during recall does not improve the network's performance. This implies that reduction in the strength of synaptic connections should occur in the presence of ACh, more than in normal conditions. We propose that two effects of ACh- enhancement of LTP and enhancement of LTD--should act together to increase the stability of the cortical network in the process of acquiring information. PMID- 7472422 TI - A serine proteinase involved in contact mediated repulsion of retinal growth cones by DRG neurites. AB - Cultured retinal ganglion cell growth cones avoid neurites extending from PNS explants. Here we characterize a growth cone collapsing activity in detergent extracts of newborn calf adrenal plasma membranes that has characteristics expected for an avoidance cue on peripheral neurites. This adrenal derived activity induces the rapid and reversible collapse of retinal growth cones grown on either of two distinct adhesion substrata, mouse laminin or the chick cell surface axonal glycoprotein G4/NgCAM. The collapsing activity is inhibited by several different types of serine proteinase inhibitors, including the irreversible inhibitor PPACK (D-phenylalanyl-prolyl-arginine chloromethyl ketone). The activity is not inhibited by the specific thrombin inhibitor, hirudin. We have named the adrenal derived collapsing activity erase. PPACK blocks the collapse of temporal retinal growth cones on contact with DRG neurites, but does not block the collapse of the same growth cones on contact with nasal retinal neurites. These results support the hypothesis that a serine proteinase on peripheral axons serves as an avoidance cue which induces contact mediated collapse of retinal growth cones. PMID- 7472423 TI - Altered sexual partner preference in male ferrets given excitotoxic lesions of the preoptic area/anterior hypothalamus. AB - Numerous experiments suggest that perinatal exposure of male vertebrates to testosterone (T), or its estrogenic metabolites, masculinizes aspects of coital function, including males' characteristic preference to seek out and mate with a female as opposed to another male conspecific. Other research has shown that this perinatal action of sex steroids also masculinizes aspects of neuronal morphology in the medial preoptic area/anterior hypothalamus (mPOA/AH). We asked whether neurons of the mPOA/AH contribute to males' preference to mate with a female. The ferret is an ideal species in which to ask this question. When tested in a T-maze after gonadectomy and treatment with estradiol benzoate (EB), female ferrets prefer to approach and receive neck grips from a stimulus male whereas males prefer to approach and neck grip an estrous female. In the minority of trials when EB-treated males approach a stimulus male, they occasionally receive a neck grip to which they display receptive postures as opposed to agonistic behaviors. In Experiment 1 castrated, EB-treated male ferrets which received bilateral infusions of the NMDA excitotoxin, quinolinic acid aimed at the dorsomedial POA/AH, preferred to approach a stimulus male significantly more often than groups of control males which either received a sham lesion, received a unilateral mPOA/AH lesion or in which bilateral infusions of quinolinic aci produced no histologically detectible excitotoxic damage to the mPOA/AH. Males with bilateral mPOA/AH lesions also displayed neck gripping on a significantly lower percentage of trials than control males when they approached the stimulus female. Ovariectomized, EB-treated female ferrets with bilateral mPOA/AH lesions, like control females, preferred to approach and receive neck grips from a stimulus male. The males used in Experiment 1 had never experienced circulating levels of T characteristic of the breeding season. Therefore, in Experiment 2 prepubertally gonadectomized males and females were treated chronically with a high dose of T propionate (TP) and tested several times with a receptive female prior to brain surgery. Males which received bilateral excitotoxic lesions of the mPOA/AH neck gripped and mounted stimulus females significantly less than control males. Again, when given EB followed by T-maze tests of partner preference, males with bilateral mPOA/AH lesions, like sham-operated female controls, preferred to approach the sexually active stimulus male significantly more often than control males, which preferred to approach the stimulus female. Our results suggest that neurons in the mPOA/AH play an important role in the integration of sensory cues which determine heterosexual partner preference in the male ferret, in addition to facilitating masculine coital performance. PMID- 7472424 TI - Glutamate-gated inhibitory currents of central pattern generator neurons in the lobster stomatogastric ganglion. AB - Inhibitory glutamatergic neurotransmission is an elemental "building block" of the oscillatory networks within the crustacean stomatogastric ganglion (STG). This study constitutes the initial characterization of glutamatergic currents in isolated STG neurons in primary culture. Superfusion of 1 mM L-glutamate evoked a current response in 45 of 65 neurons examined. The evoked current incorporated two kinetically distinct components in variable proportion: a fast desensitizing component and a slower component. The current was mediated by an outwardly rectifying conductance increase and reversed at -48.8 +/- 5.3 mV. Reducing the external chloride concentration by 50% deflected the glutamate equilibrium potential (Eglu) by +14 mV, while increasing external potassium threefold shifted Eglu by up to +6 mV. Ibotenic acid fully activated both components of the glutamate response. Saturating concentrations of glutamate completely occluded neuronal responses to ibotenic acid, indicating that ibotenic acid was activating the same receptor(s) as glutamate. Millimolar concentrations of quisqualic acid, kainate, AMPA, and NMDA each failed to evoke any response. Picrotoxin (10(-4)M) completely blocked the glutamate response. Niflumic acid (100 microM) blocked > 80% of the desensitizing component and congruent to 50% of the sustained component. Reduction or elimination of extracellular calcium did not abolish the response. This study extends the ionic and pharmacological analysis of glutamatergic conductances in STG neurons. The currents described are consistent with glutamatergic inhibitory synaptic and agonist-evoked responses previously described in situ. We discuss their pharmacology, ionic mechanisms, and functional significance. PMID- 7472426 TI - Hippocampal CA1 interneurons: an in vivo intracellular labeling study. AB - Fast spiking interneurons in the CA1 area of the dorsal hippocampus were recorded from and filled with biocytin in anesthetized rats. The full extent of their dendrites and axonal arborizations as well as their calcium binding protein content were examined. Based on the spatial extent of axon collaterals, local circuit cells (basket and O-LM neurons) and long-range cells (bistratified, trilaminar, and backprojection neurons) could be distinguished. Basket cells were immunoreactive for parvalbumin and their axon collaterals were confined to the pyramidal layer. A single basket cell contacted more than 1500 pyramidal neurons and 60 other parvalbumin-positive interneurons. Commissural stimulation directly discharged basket cells, followed by an early and late IPSPs, indicating interneuronal inhibition of basket cells. The dendrites of another local circuit neuron (O-LM) were confined to stratum oriens and it had a small but high-density axonal terminal field in stratum lacunosum-moleculare. The fastest firing cell of all interneurons was a calbindin-immunoreactive bistratified neuron with axonal targets in stratum oriens and radiatum. Two neurons with their cell bodies in the alveus innervated the CA3 region (backprojection cells), in addition to rich axon collaterals in the CA1 region. The trilaminar interneuron had axon collaterals in strata radiatum, oriens and pyramidale with its dendrites confined to stratum oriens. Commissural stimulation evoked an early EPSP-IPSP-late depolarizing potential sequence in this cell. All interneurons formed symmetric synapses with their targets at the electron microscopic level. These findings indicate that interneurons with distinct axonal targets have differential functions in shaping the physiological patterns of the CA1 network. PMID- 7472425 TI - Restricted eating with weight loss selectively decreases extracellular dopamine in the nucleus accumbens and alters dopamine response to amphetamine, morphine, and food intake. AB - Weight loss is known to alter food intake and drug self-administration, but the neural basis of this is unknown. Therefore, we studied effects of weight loss on neurochemistry of a brain mechanism involved in behavior reinforcement. In rats reduced 20-30% below normal weight, basal extracellular dopamine (DA) in the nucleus accumbens (NAC) decreased up to 50% (p < 0.01), as measured by in vivo microdialysis. No such change was observed in dorsal striatum (STR) or medial prefrontal cortex. In underweight rats, systemic amphetamine (1.5 mg/kg i.p.) transiently restored extracellular DA, but only to basal normal levels. Morphine (20 mg/kg i.p.) or a meal also increased DA, but the percent increase was significantly smaller in underweight than normal weight animals. Amphetamine infused locally by reverse dialysis in the NAC increased extracellular DA more in underweight animals than controls, suggesting that DA had accumulated in the presynaptic terminals. This was confirmed by finding significantly more DA in homogenized NAC micropunches of underweight rats. Receptor counts in micropunches and quantitative receptor autoradiography showed 3H-SCH23390 and 3H-spiperone D1- and D2-type binding in the NAC, STR, frontal cortex and hypothalamus did not change significantly. Locomotor activity was depressed suggesting that low DA release in the NAC may be related to energy conservation during weight loss. Low extracellular DA may also underlie the increase in food and drug intake typically observed in underweight animals and humans when they attempt to restore extracellular DA levels by natural or artificial means. PMID- 7472427 TI - The adult CNS retains the potential to direct region-specific differentiation of a transplanted neuronal precursor cell line. AB - The chronic survival and differentiation of the conditionally immortalized neuronal cell line, RN33B, was examined following transplantation into the adult and neonatal rat hippocampus and cerebral cortex. In clonal culture, differentiated RN33B cells express p75NTR and trkB mRNA and protein, and respond to brain-derived neurotrophic factor treatment by inducing c-fos mRNA. Transplanted cells, identified using immunohistochemistry to detect beta galactosidase expression, were seen in most animals up to 24 weeks posttransplantation (the latest time point examined). Stably integrated cells with various morphologies consistent with their transplantation site were observed. In the cerebral cortex, many RN33B cells differentiated with morphologies similar to pyramidal neurons and stellate cells. In the hippocampal formation, many RN33B cells assumed morphologies similar to pyramidal neurons characteristic of CA1 and CA3 regions, granular cell layer neurons of the dentate gyrus, and polymorphic neurons of the hilar region. Identical morphologies were observed in both adult and neonatal hosts, although a greater percentage of beta galactosidase immunoreactive cells had differentiated in the neonatal brains. These results suggest that RN33B cells have the developmental plasticity to respond to local microenvironmental signals and that the adult brain retains the capacity to direct the differentiation of neuronal precursor cells in a direction that is consistent with that of endogenous neurons. PMID- 7472428 TI - Altered nerve terminal arborization and synaptic transmission in Drosophila mutants of cell adhesion molecule fasciclin I. AB - This work investigates the role of cell adhesion molecules in development of synaptic connections and functions through a genetic approach. Fasciclin I (Fas I) is an insect glycoprotein capable of mediating homophilic cell adhesion. It has been shown that Fas I is expressed in motor nerve axons and terminals that innervate larval body-wall muscles in Drosophila. Immunohistochemical analysis of these motor nerve terminals has revealed that nerve terminal arborization, quantified by the numbers of the nerve terminal branches and varicosities, is enhanced in the null mutant fas ITE. In contrast, the number of branches and varicosities are reduced in larvae that overexpress the Fas I molecule resulting from additional copies of the fas I transgene in P(fas I+) or the chromosome duplication in Dp(fas I) mutants. Although arborization is altered, the overall stereotypical pattern of nerve terminal innervation of the body-wall muscle fibers is preserved in all the Fas I mutants examined. The voltage-clamp analysis of excitatory junctional currents (ejcs) at the neuromuscular junction indicates that the amplitude of ejcs is reduced in fas ITE, but increased in P(fas I+) and Dp(fas I) compared to that in wild-type larvae. Further electrophysiological analysis shows that the quantal content and the evoked frequency-dependent response are affected in these mutants, indicating a defective presynaptic function in addition to the anatomic abnormality. Therefore, the cell adhesion molecule Fas I may not be essential for target recognition and synaptogenesis at the larval neuromuscular junction, but may play a role in fine-turning nerve terminal arborization and possibly in modifying, directly or indirectly, development of presynaptic functions. PMID- 7472429 TI - Interleukin-6 selectively enhances the intracellular calcium response to NMDA in developing CNS neurons. AB - Increasing evidence supports a role for cytokines as chemical signals in the CNS, either under normal conditions or in the pathologic state. CNS levels of the cytokine interleukin-6 (IL-6) are known to be elevated in several diseases associated with developmental disorders and may contribute to the pathological state. To investigate the potential role of IL-6 in such disorders, neuronal effects of IL-6 were examined during development using an in vitro model system, cultured rat cerebellar granule neurons. The cultures were prepared from 8 d postnatal rat pups and exposed chronically to IL-6 (5 ng/ml) by addition to the culture medium. Neuronal effects of IL-6 were assessed by a comparison of calcium signals produced in control and IL-6 treated neurons by the glutamate receptor agonists NMDA and domoate and by K+ depolarization. IL-6 treatment significantly enhanced the response to NMDA and altered the developmental pattern of NMDA sensitivity, whereas only minor changes were observed for the response to domoate and K+. Reducing extracellular calcium and depleting intracellular stores significantly decreased the amplitude of the response to NMDA in control and IL-6 treated neurons. However, the IL-6 treated neurons were significantly more sensitive to these treatments than control neurons. These results suggest that elevated levels of IL-6 can significantly alter CNS neuron development and response to excitatory transmitters, and that IL-6 pretreatment selectively enhances the intracellular calcium responses to NMDA by altering the relative contribution of extracellular calcium influx and release of calcium from stores to the calcium signal. PMID- 7472430 TI - Modeling direction selectivity of simple cells in striate visual cortex within the framework of the canonical microcircuit. AB - Nearly all models of direction selectivity (DS) in visual cortex are based on feedforward connection schemes, where geniculate input provides all excitatory synaptic input to both pyramidal and inhibitory neurons. Feedforward inhibition then suppresses feedforward excitation for nonoptimal stimuli. Anatomically, however, the majority of asymmetric, excitatory, synaptic contacts onto cortical cells is provided by other cortical neurons, as embodied in the Canonical Microcircuit of Douglas and Martin (1991). In this view, weak geniculate input is strongly amplified in the preferred direction by the action of intracortical excitatory connections, while in the null direction inhibition reduces geniculate induced excitation. We investigate analytically and through biologically realistic computer simulations the functioning of a cortical network based on massive excitatory, cortico-cortical feedback. The behavior of this network is compared to physiological data as well as to the behavior of a purely feedforward model of DS based on nonlagged input. Our model explains a number of puzzling features of direction selective simple cells, including the small somatic input conductance changes that have been measured experimentally during stimulation in the null direction, and the persistence of DS while fully blocking inhibition in a single cell. Although the operation at the heart of our network is amplification, the network passes the linearity test of (Jagadeesh et al., 1993). We make specific predictions concerning the effect of selective blockade of cortical inhibition on the velocity-response curve. PMID- 7472431 TI - Consequences and mechanisms of spike broadening of R20 cells in Aplysia californica. AB - We studied frequency-dependent spike broadening in the two electrically coupled R20 neurons in the abdominal ganglion of Aplysia. The peptidergic R20 cells excite the R25/L25 interneurons (which trigger respiratory pumping) and inhibit the RB cells. When fired at 1-10 Hz, the duration of the falling phase of the action potential in R20 neurons increases 2-10 fold during a spike train. Spike broadening recorded from the somata of the R20 cells affected synaptic transmission to nearby follower cells. Chemically mediated synaptic output was reduced by approximately 50% when recorded trains of nonbroadened action potentials were used as command signals for a voltage-clamped R20 cell. Electrotonic EPSPs between the R20 cells, which normally facilitated by two- to fourfold during a high frequency spike train, showed no facilitation when spike broadening was prevented under voltage-clamp control. To examine the mechanism of frequency-dependent spike broadening, we applied two-electrode voltage-clamp and pharmacological techniques to the somata of R20 cells. Several voltage-gated ionic currents were isolated, including INa, a multicomponent ICa, and three K+ currents--a high threshold, fast transient A-type K+ current (IAdepol), a delayed rectifier K+ current (IK-V), and a Ca(2+)-sensitive K+ current (IK-Ca), made up of two components. The influences of different currents on spike broadening were determined by using the recorded train of gradually broadening action potentials as the command for the voltage clamp. We found the following. (1) IAdepol is the major outward current that contributes to repolarization of nonbroadened spikes. It undergoes pronounced cumulative inactivation that is a critical determinant of spike broadening. (2) Activity-dependent changes in IK-V, IK-Ca, and ICa have complex effects on the kinetics and extent of broadening. (3) The time integral of ICa during individual action potentials increases approximately threefold during spike broadening. PMID- 7472432 TI - Elemental composition and water content of rat optic nerve myelinated axons and glial cells: effects of in vitro anoxia and reoxygenation. AB - Electron probe x-ray microanalysis was used to measure water content and concentrations (mmol/kg dry weight) of elements (Na, P, S, Cl, K, Ca, and Mg) in myelinated axons and glial cells of rat optic nerve exposed to in vitro anoxia and reoxygenation. In response to anoxia, large, medium, and small diameter fibers exhibited an early (5 min) and progressive loss of Na and K regulation which culminated (60 min) in severe depletion of respective transmembrane gradients. As axoplasmic Na levels increased during anoxic exposure, a parallel rise in Ca content was noted. For all axons, mean water content decreased progressively during the initial 10 min of anoxia and then returned toward normal values as anoxia continued. Analyses of mitochondrial areas revealed a similar pattern of elemental disruption except that Ca concentrations rose more rapidly during anoxia. Following 60 min of postanoxia reoxygenation, the majority of larger fibers displayed little evidence of recovery, whereas a subpopulation of small axons exhibited a trend toward restoration of normal elemental composition. Glial cells and myelin were only modestly affected by anoxia and subsequent reoxygenation. Thus, anoxic injury of CNS axons is associated with characteristic changes in axoplasmic distributions of Na, K, and Ca. The magnitude and temporal patterns of elemental Na and Ca disruption are consistent with reversal of Na(+) Ca2+ exchange and subsequent Ca entry (Stys et al., 1992). During reoxygenation, elemental deregulation continues for most CNS fibers, although a subpopulation of small axons appears to be capable of recovery. PMID- 7472433 TI - Localization of thioredoxin in the rat brain and functional implications. AB - The immunoreactivity for thioredoxin, which catalyzes protein disulfide reductions, has previously been shown to exist in nerve cells and their axons. Here we demonstrate the localization of thioredoxin mRNA as revealed by in situ hybridization in the rat brain. The gene is expressed in nerve cells of a variety of brain regions, for example, the cerebral cortex, the piriform cortex, the medial preoptic area, the CA3/CA4 region of the hippocampal formation, the dentate gyrus, the paraventricular nucleus of the hypothalamus, the arcuate nucleus, the substantia nigra pars compacta, the locus coeruleus, the ependyma of the 4th ventricle, and the epithelial cells of the choroid plexus. This distribution implicates an important function in nerve cell metabolism, especially in regions with high energy demands and indicates a role of the choroid plexus in nerve cell protection from environmental influences. It was found that after mechanical injury induced by partial unilateral hemitransection the thioredoxin mRNA expression is upregulated in the lesioned area and spreads to the cortical hemispheres at the lesioned level. This induction suggests a function of thioredoxin in the regeneration machinery of the brain following mechanical injury and oxidative stress. PMID- 7472434 TI - TOAD-64, a gene expressed early in neuronal differentiation in the rat, is related to unc-33, a C. elegans gene involved in axon outgrowth. AB - Using two-dimensional gel electrophoresis we previously identified membrane associated proteins that are upregulated over the course of neurogenesis. One of these, TOAD-64 (Turned On After Division, 64 kDa), is expressed immediately after neuronal birth and is dramatically downregulated in the adult. The gene encoding TOAD-64 has now been cloned, and its sequence shows homology to the unc-33 gene from C. elegans, mutations in which lead to aberrations in axon outgrowth. Northern and in situ hybridization show that TOAD-64 mRNA is enriched in the nervous system and is developmentally regulated in parallel with the protein. The expression of the TOAD-64 protein and gene coincident with initial neuronal differentiation and the downregulation when the majority of axon growth is complete suggests a role in axon elaboration. Three additional lines of evidence support this possibility: TOAD-64 is upregulated following neuronal induction of P19 and PC12 cells; the protein is found in lamellipodia and filopodia of growth cones; and axotomy of the sciatic nerve induces reexpression. While the sequence of TOAD-64 lacks a signal sequence and therefore is likely to encode a cytoplasmic protein, biochemical experiments demonstrate that the protein is tightly, but noncovalently, associated with membranes. The data presented here suggest that TOAD-64 could be a central element in the machinery underlying axonal outgrowth and pathfinding, perhaps playing a role in the signal transduction processes that permit growing axons to choose correct routes and targets. PMID- 7472435 TI - Tissue- and age-specific expression patterns of alternatively spliced agrin mRNA transcripts in embryonic rat suggest novel developmental roles. AB - Agrin is an extracellular matrix protein that mediates the nerve-induced clustering of nicotinic acetylcholine receptors on target muscle cells, and thus plays a key role in development of the neuromuscular synapse. Alternative exon usage within the rat agrin gene predicts numerous protein isoforms, which differ by the inclusion or exclusion of small inserts at three sites in the C-terminal half of the molecule; the insert status at two of these sites, termed Y and Z, profoundly influences the acetylcholine receptor clustering activity. We have examined the cellular expression patterns of agrin messenger RNA transcripts during rat embryogenesis by in situ hybridization with isoform-specific probes. Six 36-mer oligonucleotide probes were designed to distinguish between mRNA isoforms at either the Y site: the encoded protein contains either no insert (Y0) or a 4-amino acid insert (Y4), or the Z site: the encoded protein contains either no insert or one of 8 (Z8), 11 (Z11), or 19 (Z19) amino acids. Strikingly different expression patterns were observed for the individual Y- and Z-site encoding messages. While optional exon usage predicts the possibility of eight different agrin isoforms at the two splice sites, we detected only four isoforms in vivo: Y4Z0, Y0Z0, Y4Z8, and Y4Z19. The Y4Z0 transcript, which comprised the majority of the agrin expressed, was localized exclusively to nervous tissue and exhibited a distribution profile suggestive of a potential role in neurogenesis and/or neural differentiation. From embryonic day 13 to birth, Y4Z0 was found in mitotic ventricular zones, spinal, cranial, and sympathetic ganglia, and diffusely throughout the brain. In contrast, Y0Z0 was not expressed in neurons, but specifically labeled capillary endothelial cells within the developing nervous system. Y4Z8 and Y4Z19, the forms most active in acetylcholine receptor aggregation, were expressed at low levels only in spinal and brainstem motor neurons; Z19 expression declined from embryonic day 15 to adulthood, whereas Z8 expression increased slightly during this period. Transcripts encoding the Z11 insert could not be detected. These data suggest potential novel biological roles for agrin beyond that originally proposed in synapse formation. PMID- 7472436 TI - Glutamate receptors in the nucleus accumbens shell control feeding behavior via the lateral hypothalamus. AB - The nucleus accumbens in a brain region considered to be important in the regulation of appetitive behavior and reinforcement. The accumbens receives afferent input from corticolimbic and thalamic structures, which is primarily coded by excitatory amino acids (EAAs). The present studies investigated the role of EAA input to the nucleus accumbens in feeding behavior in rats, in two recently characterized subregions of the accumbens, the "core" and "shell". In the first series of experiments, it was shown that blockade of alpha-amino-3 hydroxy-5-methylisoxazole-4-propionic acid (AMPA) and kainate glutamate receptors in the medial part of the accumbens, corresponding to the medial shell subregion, resulted in a pronounced feeding response. Bilateral microinfusion of 6,7 dinitroquinoxaline-2,3-dione (DNQX, 0.25-0.75 micrograms/0.5 microliters), 6 cyano-7-nitroquinoxaline (CNQX, 0.75-1.5 micrograms), and 2,3-dihydroxy-6-nitro-7 sulfamoylbenzo-(F) quinoxaline (NBQX, 0.2-1.0 micrograms) markedly stimulated food intake immediately following infusion, in a dose-dependent manner. Infusion of DNQX into the central accumbens region, corresponding to the core, did not elicit feeding. Infusion of the NMDA antagonists 2-amino-5-phosphonopentanoic acid (AP-5) and MK-801 (dizocilpine maleate) did not elicit feeding in either region. The feeding response to DNQX was blocked by local coinfusion of AMPA. Systemic pretreatment with naltrexone (5 mg/kg) had no effect on the DNQX-feeding response; however, prior systemic administration of both D-1 and D-2 antagonists reduced the response by half, suggesting a modulatory role for dopamine in the response. Moreover, the feeding response was completely inhibited by concurrent infusion of the GABAA agonist muscimol (10, 25 ng) into the lateral hypothalamus, a major projection area of the accumbens shell. These findings demonstrate a selective role for non-NMDA receptors in the nucleus accumbens shell in ingestive behavior, and suggest an important functional link between two major brain regions involved in reward, the nucleus accumbens and lateral hypothalamus. PMID- 7472437 TI - Forebrain contribution to the induction of a cellular correlate of conditioned taste aversion in the nucleus of the solitary tract. AB - A conditioned taste aversion (CTA) is a form of classical conditioning in which animals avoid a taste (conditioned stimulus; CS) which has been previously paired with a treatment (unconditioned stimulus; US) that produces transient illness. Recently, a reliable cellular correlate of the behavioral expression of a CTA was identified using c-Fos immunostaining as a marker of neuronal activation. Exposure to a saccharin solution (CS) which had previously been paired with lithium chloride (LiCl; US) induced significant c-Fos-like immunoreactivity (c FLI) in the intermediate zone of the nucleus of the solitary tract (NTS), a response that was quite similar to that displayed following administration of LiCl alone. The present studies employed a variant of the chronic decerebrate rat preparation to explore whether circuitry intrinsic to the brainstem is sufficient for the induction of c-FLI in both as an unconditioned response to the LiCl and as a conditioned response to the saccharin. Using chronic hemidecerebrate rats, which have a unilateral brain transection at the level of the superior colliculus, we found that the unconditioned c-FLI to LiCI was unaltered by the transection, while the conditioned expression of c-FLI to the CS taste was evident only on the side of the NTS which retained neural connections with the forebrain. These findings strongly implicate forebrain input in this cellular correlate of CTA learning and also indicate that the pathways mediating the response to the US (LiCl) and the CS (saccharin) differ. PMID- 7472439 TI - kappa-opioid receptor agonists modulate excitatory transmission in substantia gelatinosa neurons of the rat spinal cord. AB - This study examined the effects of selective activation of kappa 1-opioid receptors on excitatory transmission in substantia gelatinosa (SG) using intracellular recordings from SG neurons in transverse slices of the young rat lumbar spinal cord. Monosynaptic and polysynaptic excitatory postsynaptic potentials (EPSPs) were evoked by orthodromic electrical stimulation of A delta or C primary afferent fibers in the dorsal root after blocking inhibitory inputs with bicuculline and strychnine, NMDA receptors with D-2-amino-5-phosphonovaleric acid and mu- and delta-opioid receptors with CTAP and ICI 174,864, respectively. Bath application of dynorphin A1-17 or U-69, 593 caused dual modulation of the peak amplitude of presumed monosynaptic AMPA receptor-mediated EPSPs, decreasing synaptic potentials at nanomolar concentrations in a majority of SG cells examined (dynorphin, 63%; U-69,593, 91%), and increasing EPSPs at micromolar concentrations. Only the inhibitory action of dynorphin A1-17 was consistently and completely blocked by norbinaltorphimine (nor-BNI). Since U-69,593 and nor BNI are selective for the kappa 1-opioid receptors, the depression of EPSPs is likely to be mediated by the kappa1-opioid receptors. Under conditions of blockade of synaptic transmission with TTX and mu-and delta-opioid receptors, dynorphin A1-17 and U-69,593 hyperpolarize most of SG neurons and decrease their membrane input resistance, the finding suggesting that direct interaction of kappa-agonists with a postsynaptic receptor is likely explanation for the inhibition of EPSPs. However, in some SG cells, the inhibition of EPSPs appears to be of presynaptic origin since dynorphin A1-17 and U-69,593 did depress the EPSPs in the absence of changes in passive membrane properties. Rp-cAMPS, a membrane permeant potent competitive inhibitor of cAMP-activated protein kinase, prevented the depressant effect of dynorphin A 1-17. This finding suggested a possibility that dynorphin A1-17, acting through a decrease in intracellular cyclic AMP levels, can reduce the synaptic responses of SG neurons. These results provide the first electrophysiological demonstration that the activation of kappa 1-opioid receptors inhibits AMPA receptor-mediated primary afferent neurotransmission in the substantia gelatinosa of the young rat spinal cord. This effect may mediate the ability of kappa-receptor agonists to produce antinociception. PMID- 7472438 TI - Developmental and regional expression of multifunctional Ca2+/calmodulin dependent protein kinase isoforms in rat brain. AB - Multifunctional Ca2+/calmodulin-dependent protein kinase (CaM kinase) participates in diverse calcium signaling pathways in neurons. The alpha- and beta-CaM kinase isoforms are neuron-specific and highly abundant in rat brain. The variable domain of CaM kinase is a potential site for the generation of isoform diversity by alternative spicing of its N- and/or C-terminal segments. We used specific PCR primers which span the variable domain of either alpha- or beta CaM kinase and isolated three new isoforms from rat brain, namely alpha B-, beta e- and beta'e-CaM kinase. alpha beta-CaM kinase contains 11 amino acids, likely inserted by alternative splicing, at the C-terminal segment of the variable domain. This insertion introduces a nuclear localization signal (NLS) that targets alpha B-CaM kinase to the nucleus of transfected cells; alpha-CaM kinase is excluded from the nucleus. The mRNA and the protein corresponding to this isoform are detected only in the diencephalon/midbrain regions. We have also identified two alternatively spliced isoforms of beta-CaM kinase that lack the 24 amino acid sequence at the N-terminal segment of the variable domain. Alternative splicing of these two isoforms occurs with a three base pair shift of the 3' splice site. Our analysis shows that these new beta-CaM kinase isoforms are expressed primarily in early developmental stages, and we therefore term them beta e - (embryonic) and beta' e-CaM kinase. Recombinant alpha B-, beta e and beta' e-CaM kinase expressed in COS-7 cells exhibit characteristic Ca2+/calmodulin-dependent protein kinase activity and autophosphorylation. PMID- 7472440 TI - The growth of the axon is not dependent upon net microtubule assembly at its distal tip. AB - Although there is agreement that the net addition of new microtubule polymer to the axon is required for its growth, controversy exists concerning the principal site in the neuron where this occurs. Some models hold that microtubule polymer is assembled within the cell body and translocated down the axon, while others hold that the net addition of polymer occurs at the distal tip of the axon. The foundation for the latter idea was a study in which anti-microtubule drugs were applied topically for 30 min to discrete regions of cultured sensory neurons (Bamburg et al., 1986). The axon continued to grow when the drugs were applied to the cell body, but stopped growing when the drugs were applied to the distal tip of the axon. Assuming that the sole action of the drug was to inhibit microtubule assembly, many workers have interpreted these findings as indicating that the growth of the axon requires net microtubule assembly at its distal tip. We repeated these experiments using a broader range of drug treatments, and evaluated using electron microscopy the effects of these treatments on microtubule levels. Our results indicate that the previous drug treatments went beyond inhibiting microtubule assembly, and also caused substantial microtubule disassembly. When the drug regime was altered so as to induce lower levels of microtubule disassembly in the distal region of the axon, the axon continued to grow. These results indicate that the growth of the axon is not dependent upon net microtubule assembly at its distal tip. PMID- 7472441 TI - The mechanisms of generation and propagation of synchronized bursting in developing networks of cortical neurons. AB - The characteristics and mechanisms of synchronized firing in developing networks of cultured cortical neurons were studied using multisite recording through planar electrode arrays (PEAs). With maturation of the network (from 3 to 40 d after plating), the frequency and propagation velocity of bursts increased markedly (approximately from 0.01 to 0.5 Hz and from 5 to 100 mm/sec, respectively), and the sensitivity to extracellular magnesium concentration (0-10 mM) decreased. The source of spontaneous bursts, estimated from the relative delay of onset of activity between electrodes, varied randomly with each burst. Physical separation of synchronously bursting networks into several parts using an ultraviolet laser, divided synchronous bursting into different frequencies and phases in each part. Focal stimulation through the PEA was effective at multiple sites in eliciting bursts, which propagated over the network from the site of stimulation. Stimulated bursts exhibited both an absolute refractory period and a relative refractory period, in which partially propagating bursts could be elicited. Periodic electrical stimulation (at 1 to 30 sec intervals) produced slower propagation velocities and smaller numbers of spikes per burst at shorter stimulation intervals. These results suggest that the generation and propagation of spontaneous synchronous bursts in cultured cortical neurons is governed by the level of spontaneous presynaptic firing, by the degree of connectivity of the network, and by a distributed balance between excitation and recovery processes. PMID- 7472442 TI - Impaired fear conditioning following unilateral temporal lobectomy in humans. AB - Classical fear conditioning was used in the present study as a model for investigating emotional learning and memory in human subjects with lesions to the medial temporal lobe. Animal studies have revealed a critical role for medial temporal lobe structures, particularly the amygdala, in simple and complex associative emotional responding. Whether these structures perform similar functions in humans is unknown. On both simple and conditional discrimination tasks, unilateral temporal lobectomy subjects showed impaired conditioned response acquisition relative to control subjects. This impairment could not be accounted for by deficits in nonassociative sensory or autonomic performance factors, or by differences in declarative memory for the experimental parameters. These results show that temporal lobe structures in humans, as in other mammals, are important components in an emotional memory network. PMID- 7472443 TI - Changes in neurotrophic factor expression and receptor activation following exposure of hippocampal neuron/astrocyte cocultures to kainic acid. AB - Neurotrophic factor expression in the adult mammalian CNS is largely neuronal. However, upon traumatic injury reactive astrocytes express a number of neurotrophic factors including ciliary neurotrophic factor (CNTF), fibroblast growth factor (FGF), and NGF. In this study, we examined whether the upregulation of neurotrophic factors in reactive astrocytes and cultured astrocytes is a consequence of separation from their neuronal counterparts, and whether neurotrophic factor levels can be regulated by placing astrocytes into coculture with neurons. We show that reintroduction of rat hippocampal neurons to rat hippocampal astrocytes in vitro leads to a time dependent downregulation in astrocytes of the neurotrophic factors CNTF, NGF, and neurotrophin 3 (NT-3). In contrast, brain-derived neurotrophic factor (BDNF) mRNA, which is only expressed in neurons in these cultures is slightly increased. Once neurotrophic factor levels in cocultures had reached a steady state in the neuron/glia cocultures, we initiated a traumatic event with the excitotoxin kainic acid. BDNF protein was rapidly upregulated within 24 hr after lesion, whereas CNTF protein upregulation was delayed reaching maximal levels by 3 d. Despite the endogenous upregulation of both of these trophic factors, no activation of their respective receptors, as measured by tyrosine phosphorylation, was detectable following kainate administration. However, following addition of exogenous CNTF at any time point up to 24 hr after kainate administration, the beta components of the CNTF receptor (LIFR beta and gp130) could be phosphorylated. Furthermore, although activation of neuronal LIFR beta and gp130 by exogenous CNTF declined during the period of neuronal death, these receptors reappeared on astrocytes and could be activated by CNTF. In contrast, phosphorylation of TrkB by exogenous BDNF was undetectable by 24 hr and could not be reactivated after this point. These data suggest that the intimate association of astrocytes and neurons in the CNS serves to suppress astrocyte-derived neurotrophic factor expression and that neuronal loss leads to a derepression of neurotrophic factor synthesis in astrocytes. However, the upregulation of endogenous BDNF and CNTF observed after excitotoxic lesion in this culture model are insufficient to activate signal transduction and protect against neuronal loss. PMID- 7472444 TI - Morphological evidence for the sprouting of inhibitory commissural fibers in response to the lesion of the excitatory entorhinal input to the rat dentate gyrus. AB - Recently a commissural fiber projection that terminates in the outer molecular layer of the fascia dentata was described in normal rats (Deller et al., 1995). In the present article, Phaseolus vulgaris leucoagglutinin (PHAL) tracing was used to analyze the contribution of this previously unknown projection to the commissural sprouting response after entorhinal cortex lesion. Rats 4-8 weeks after unilateral entorhinal lesion received a single PHAL deposit into the hilus of the fascia dentata contralateral to the lesion side. Unlesioned control animals received a similar PHAL deposit. The degree of axonal arborization and the bouton density per axon length were determined for individual PHAL-labeled commissural axons to the outer molecular layer of the dentate gyrus. A significant increase in both parameters was observed in the lesioned group. The PHAL-labeled commissural fibers established symmetric synapses in the denervated outer molecular layer. Postembedding immunocytochemistry indicated that some of these sprouting commissural fibers are GABAergic. Our findings provide morphological evidence for lamina-specific sprouting of an inhibitory commissural projection that normally terminates in the outer molecular layer. This suggests that inhibitory fibers participate in the replacement of the excitatory perforant pathway after entorhinal lesion. PMID- 7472445 TI - Multiple presynaptic metabotropic glutamate receptors modulate excitatory and inhibitory synaptic transmission in hippocampal area CA1. AB - The metabotropic glutamate receptors (mGluRs) have many important roles in regulation of neuronal excitability and synaptic transmission. In hippocampal area CA1, activation of mGluRs can reduce both excitatory and inhibitory synaptic transmission. The conventional view is that the presynaptic effects are mediated by L-2-amino-4-phosphonobutyric acid (L-AP4)-sensitive, or group III mGluRs (mGluR4, mGluR6, mGluR7, mGluR8). However, some studies suggest that other mGluR subtypes may also be involved in regulation of excitatory and inhibitory synaptic transmission in area CA1. We have found that two pharmacologically distinct presynaptic receptors are involved in the depression of excitatory transmission at the Schaffer collateral--CA1 synapse. Consistent with previous studies, one receptor subtype is an L-AP4-sensitive receptor that is pharmacologically similar to mGluR4 or mGluR7. However, we have found that a second mGluR subtype, which is pharmacologically similar to mGluR1 and mGluR5 (group I mGluRs), can also reduce excitatory synaptic transmission in area CA1. Analysis of effects of agonists of these two receptors on miniature EPSCs and paired-pulse facilitation suggest that both receptors are localized presynaptically. It is also shown that the mGluR that reduces transmission at inhibitory synapses in area CA1 is presynaptically localized, is insensitive to L-AP4, and is sensitive to agonists selective for mGluR1 and mGluR5. PMID- 7472446 TI - Development of glycinergic and glutamatergic synaptic transmission in the auditory brainstem of perinatal rats. AB - In contrast to our knowledge about the anatomical development of the mammalian central auditory system, the development of its physiological properties is still poorly understood. In order to better understand the physiological properties of the developing mammalian auditory brainstem, we made intracellular recordings in brainstem slices from perinatal rats to examine synaptic transmission in the superior olivary complex, the first binaural station in the ascending auditory pathway. We concentrated on neurons in the lateral superior olive (LSO), which in adults, are excited from the ipsilateral side and inhibited from the contralateral side. Already at embryonic day (E) 18, when axon collaterals begin to invade the LSO anlage, synaptic potentials could be evoked from ipsilateral, as well as from contralateral inputs. Ipsilaterally elicited PSPs were always depolarizing, regardless of age. They had a positive reversal potential and could be completely blocked by the non-NMDA glutamate receptor antagonist CNQX. In contrast, contralaterally elicited PSPs were depolarizing from E18-P4, yet they turned into "adult-like," hyperpolarizing PSPs after P8. Their reversal potential shifted dramatically from -21.6 +/- 17.7 mV (E18-P0) to -73.0 +/- 7.1 mV (P10). Regardless of their polarity, contralaterally elicited PSPs were reversibly blocked by the glycine receptor antagonist strychnine. Bath application of glycine and its agonist beta-alanine further confirmed the transitory depolarizing action of glycine in the auditory brainstem. Since the transient excitatory behavior of glycine occurs during a period during which glycinergic synaptic connections in the LSO are refined by activity-dependent mechanisms, glycinergic excitation might be a mechanism by which synaptic rearrangement in the contralateral inhibitory pathway is accomplished. PMID- 7472447 TI - Saccade target selection in frontal eye field of macaque. I. Visual and premovement activation. AB - We investigated how the brain selects the targets for eye movements, a process in which the outcome of visual processing is converted into guided action. Macaque monkeys were trained to make a saccade to fixate a salient target presented either alone or with multiple distractors during visual search. Neural activity was recorded in the frontal eye field, a cortical area at the interface of visual processing and eye movement production. Neurons discharging after stimulus presentation and before saccade initiation were analyzed. The initial visual response of frontal eye field neurons was modulated by the presence of multiple stimuli and by whether a saccade was going to be produced, but the initial visual response did not discriminate the target of the search array from the distractors. In the latent period before saccade initiation, the activity of most visually responsive cells evolved to signal the location of the target. Target selection occurred through suppression of distractor evoked activity contingent on the location of the target relative to the receptive field. The evolution of a signal specifying the location of the salient target could be dissociated from saccade initiation in some cells and could occur even when fixation was maintained. Neural activity in the frontal eye fields may participate in or be the product of the decision process guiding eye movements. PMID- 7472448 TI - Repeated exposure to one song leads to a rapid and persistent decline in an immediate early gene's response to that song in zebra finch telencephalon. AB - Conspecific song induces a prompt increase in the expression of the zenk gene in the caudo-medial neostriatum (NCM), a part of the auditory telencephalon of songbirds. To test the hypothesis that zenk gene induction in NCM is related to the acquisition of new song-related memories, we presented adult male zebra finches with repeated playbacks of one song. In response, zenk mRNA levels in NCM increased for the first 30 min, but then declined back to baseline levels despite continued stimulation with the same song. When a novel song was then introduced, however, a full zenk response was triggered once again. Even when a full day had passed between the last exposure to a song and the testing period, the song that had been rendered "familiar" by repetition caused no zenk response, whereas a novel song caused a full response. Quantitative analysis indicates that individual cells in NCM must have undergone a selective loss in their zenk responsiveness to the repeated song, while still maintaining their ability to respond to the novel song. These results support the hypothesis that the induction of zenk is related to the formation of long-term memories. Analysis of the stimulus-specific modulation of zenk responses, coupled with appropriate behavioral assays, should provide insight into neural mechanisms responsible for the discrimination and storage of complex perceptual information. PMID- 7472449 TI - Recovery of tail-elicited siphon-withdrawal reflex following unilateral axonal injury is associated with ipsi- and contralateral changes in gene expression in Aplysia californica. AB - Behavioral, cellular and molecular changes were examined following axonal injury in the marine mollusc Aplysia californica. Unilateral nerve injury was performed by crushing the pleural-pedal connective and the peripheral pedal nerves innervating one side of the posterior body wall and the tail. The injury procedure severs the axons of the pleural sensory neurons resulting in the blockade of the tail-elicited siphon-withdrawal reflex. Partial reflex recovery is observed within 3 d and reaches 50% of the pretest value by six weeks postinjury. Retrograde staining of injured nerves combined with electrophysiological recordings from siphon motor neurons show that axons can regenerate through the crushed site and reconnect with the tail by three weeks postinjury. Moreover, the behavioral and electrophysiological measurements suggest that the contralateral sensory neurons may contribute to the early recovery of the siphon-withdrawal reflex. The levels of mRNAs coding for actin and calreticulin are elevated while the mRNAs coding for intermediate filament protein, sensorin A, FMRFamide are reduced in the ipsilateral pleural ganglia as detected by Northern blots. In the contralateral pleural ganglia, the levels of mRNAs coding for actin, sensorin A and FMRFamide are elevated. These molecular changes in both the ipsi- and contralateral sides are consistent with the hypothesis that both sides are participating in the behavioral recovery following unilateral axonal injury. PMID- 7472450 TI - A comparison of the role of dynorphin in the hippocampal mossy fiber pathway in guinea pig and rat. AB - Several behavioral studies in rat (Gallagher, 1988) have suggested that opioids in the hippocampus could play an important role in learning and memory. However, in this species, very few reports specifically address the issue of physiological actions of opioids released by the mossy fibers which constitute the principal source of dynorphin and enkephalin in the hippocampus. In the guinea pig high frequency stimulation of mossy fibers causes a transient heterosynaptic inhibition of neighboring mossy fibers (Weisskopf et al., 1993) or perforant path synapses in the dentate (Wagner et al., 1993), which is mediated by the synaptic release of dynorphin that activates presynaptic kappa receptors. We show here that neither exogenous nor endogenous dynorphin affect mossy fiber excitatory postsynaptic potentials in the Sprague-Dawley rat, which is consistent with the finding that kappa receptor binding in the mossy fiber termination zone is dense in the guinea pig and sparse in this rat. More surprisingly, although kappa receptor binding is found in the rat dentate gyrus molecular layer and in the CA3 pyramidal cell layer, dynorphin had no action on perforant path field responses, somatic potassium currents or evoked monosynaptic inhibitory postsynaptic currents in CA3 cells. This lack of action appears to be an exception among rodents as dynorphin significantly inhibited mossy fiber responses in the hamster, mouse, and even another strain of rat, Long-Evans. Unlike the kappa mediated actions, the mu opioid receptor agonist DAMGO inhibited Sprague-Dawley mossy fiber responses, as it does in guinea pig. In contrast to other investigators, however, we found that the opioid receptor antagonist naloxone had no effect on Sprague-Dawley mossy fiber LTP. PMID- 7472451 TI - Coupling potentials in CA1 neurons during calcium-free-induced field burst activity. AB - Small amplitude depolarizations (fast prepotentials, spikelets) recorded in mammalian neurons are thought to represent either dendritic action potentials or presynaptic action potentials attenuated by gap junctions. We have used whole cell recordings in an in vitro calcium-free model of epilepsy to record spikelets from CA1 neurons of the rat hippocampus. It was found that spikelet appearance was closely correlated with the occurrence of dye coupling between pyramidal neurons, indicating that both phenomena share a common substrate. Spikelets were characterized according to waveform (amplitude and shape) and temporal occurrence. Spikelet amplitudes were found to be invariant with neuronal membrane potential, and their pattern of occurrence was indistinguishable from patterns of action potential firing in these cells. Voltage and current recordings revealed a spikelet waveform that was usually biphasic, comprised of a rapid depolarization followed by a slower hyperpolarization. Numerical differentiation of spike bursts resulted in waveforms similar to recorded spikelet sequences, while numerical integration of spikelets yielded waveforms that were indistinguishable from action potentials. Modification of spikelet waveforms by the potassium channel blocker tetraethylammonium chloride suggests that spikelets may arise from both resistive and capacitive transmission of presynaptic action potentials. Intracellular alkalinization and acidification brought about by perfusion with NH4Cl caused changes in spikelet frequency, consistent with reported alterations of field burst activity in this model of epilepsy. These results suggest that spikelets result from gap junctional communication, and may be important determinants of neuronal activity during seizure-like activity. PMID- 7472452 TI - GABA antagonists differentiate between recombinant GABAA/benzodiazepine receptor subtypes. AB - Seventeen rat GABAA receptor subtypes were transiently expressed in the human embryonic kidney 293 cell line from alpha 1, alpha 2, alpha 3, alpha 5, or alpha 6 variants with any of the three beta subunits and gamma 2S or gamma 3. We obtained fingerprints in the form of subtype characteristic concentration response curves of 35S-TBPS binding to GABA and the GABAA antagonists SR 95531 and bicuculline. alpha 3 beta 3 gamma 2S/3 and alpha 5 beta 3 gamma 2S/3 containing receptors effectively recognized 35S-TBPS but not when beta 3 was replaced by the beta 1 or beta 2 subunit. This indicates a specific interaction of alpha and beta variants to form high-affinity 35S-TBPS binding sites. At low levels GABA allosterically increased 35S-TBPS binding to all receptors with the concentration and magnitude depending on the subunit combination. Exchange of the beta variant did not alter the concentration-response curves for alpha 1 and alpha 6 containing receptors but did so for alpha 2 containing receptors. alpha 2 beta 3 gamma 3 receptors displayed strong GABA-induced stimulation of 35S-TBPS binding, whereas binding to alpha 2 beta 3 gamma 3 receptors was marginally increased. SR 95531 and bicuculline decreased 35S-TBPS binding to all gamma 3 containing receptors. In addition, bicuculline was effective on alpha 1 beta x gamma 2 receptors. SR 95531 was threefold more potent than bicuculline in reversing GABA-induced modulation of 35S-TBPS binding in most receptor types, but was 30-fold more potent on alpha 2 beta 1 gamma 3 and alpha 6 beta 1 gamma 2S receptors.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7472453 TI - Axonal regeneration and physiological activity following transection and immunological disruption of myelin within the hatchling chick spinal cord. AB - Transections of the chicken spinal cord after the developmental onset of myelination at embryonic day (E) 13 results in little or no functional regeneration. However, intraspinal injection of serum complement proteins with complement-binding GalC or 04 antibodies between E9-E12 results in a delay of the onset of myelination until E17. A subsequent transection of the spinal cord as late as E15 (i.e., during the normal restrictive period for repair) results in neuroanatomical regeneration and functional recovery. Utilizing a similar immunological protocol, we evoked a transient alteration of myelin structure in the posthatching (P) chicken spinal cord, characterized by widespread "unravelling" of myelin sheaths and a loss of MBP immunoreactivity (myelin disruption). Myelin repair began within 7 d of cessation of the myelin disruption protocol. Long term disruption of thoracic spinal cord myelin was initiated after a P2-P10 thoracic transection and maintained for > 14 d by intra-spinal infusion of serum complement proteins plus complement-binding GalC or 04 antibodies. Fourteen to 28 d later, retrograde tract tracing experiments, including double labeling protocols, indicated that approximately 6-19% of the brainstem-spinal projections had regenerated across the transection site to lumbar levels. Even though voluntary locomotion was not observed after recovery, focal electrical stimulation of identified brainstem locomotor regions evoked peripheral nerve activity in paralyzed preparations, as well as leg muscle activity patterns typical of stepping in unparalyzed animals. This indicated that a transient alteration of myelin structure in the injured adult avian spinal cord facilitated brainstem-spinal axonal regrowth resulting in functional synaptogenesis with target neurons. PMID- 7472454 TI - Genetic and pharmacological analysis of neurotransmitters controlling egg laying in C. elegans. AB - We have investigated the neurotransmitters used to control egg-laying in C. elegans. Previous studies suggested that 5-HT released by the HSN motor neurons stimulates egg laying, and that tricyclic antidepressants potentiate egg laying by blocking reuptake of 5-HT by the HSN neurons. We report studies of the wild type and a mutant that lacks detectable 5-HT that suggest 5-HT is not required for egg-laying. Furthermore, we find that ACh is required for egg laying in response to 5-HT, suggesting that 5-HT is not sufficient to activate egg laying. The dominant egl-2(n693) mutation, which causes animals to lay eggs in response to tricyclics but not 5-HT, also conflicts with the model for egg laying. Experiments in which the HSN neurons or 5-HT are removed from egl-2 animals indicate that the action of tricyclics cannot be explained by a block of 5-HT reuptake. We find that D2 family dopamine antagonists can also induce egg laying in egl-2(n693) mutants, and that dopamine inhibits egg laying in the wild type. These results suggest that dominant egl-2 mutations activate an inhibitory dopaminergic pathway that can be blocked by tricyclics and D2 antagonists. We also find that these drugs stimulate egg laying in mutants lacking 5-HT or the HSN neurons, consistent with a target on the egg-laying muscles. In contrast to tricyclics, fluoxetine and other selective 5-HT reuptake inhibitors appear to be specific for 5-HT reuptake in C. elegans egg laying. PMID- 7472455 TI - Strategies for the generation of neuronal diversity in the developing central nervous system. AB - During development, the neural tube produces a large diversity of neuronal phenotypes from a morphologically homogeneous pool of precursor cells. In recent years, the cellular and molecular mechanisms by which specific types of neurons are generated have been explored, in the hope of discovering features common to development throughout the nervous system. This article focuses on three strategies employed by the CNS to generate distinct classes of neuronal phenotypes during development: dorsal-ventral polarization in the spinal cord, segmentation in the hindbrain, and a lamination in the cerebral cortex. The mechanisms for neurogenesis exemplified by these three strategies range from a relatively rigid, cell lineage-dependent specification with a high degree of subservance to early patterns of gene expression, to inductions and cell-cell interactions that determine cell fates more flexibly. PMID- 7472456 TI - Excitotoxic degeneration is initiated at non-random sites in cultured rat cerebellar neurons. AB - Prolonged stimulation of cultured cerebellar neurons by kainic acid (KA) leads to death of neurons first evident from the swelling of soma and neurites. Stimulation is accompanied by increases in [Ca2+]i and [Na+]i as monitored using digital imaging microfluorimetry. "Blebs" tended to form on neurites with the highest increases in [Ca2+]i. Points of Ca2+ entry into neurites via glutamate receptor-gated channels predicted where approximately 80% of blebs would form tens of minutes later. These sites were close to neurite intersections where there was a high likelihood of synaptic contacts and were enriched in mitochondria as revealed by rhodamine 123 staining. Ca2+, but not Na+ entry, produced a loss of mitochondrial potential. Prolonged KA, but not 50K, applications could fully dissipate the neuronal Na+ gradient. Recovery of resting [Na+]i was delayed by Ca2+ loading. We propose that blebs form at certain synaptic regions due to localized ionic fluxes and local Ca2+ overloading. Increased [Ca2+]i may hamper restoration of normal [Na+]i permitting local osmotic swelling as well as activation of Ca(2+)-dependent enzymes and other processes. Na+ may slow, block, or reverse Na/Ca exchange and enhance swelling. These conditions could not be reproduced by global changes in ion concentrations produced by Ca2+ or Na+ ionophores. The earliest stages of excitotoxicity thus appear to be manifestations of localized disruptions of ionic homeostasis mediated by Ca2+ overload and Na+ influx. PMID- 7472457 TI - Pronounced cellular diversity and extrasynaptic location of nicotinic acetylcholine receptor subunit immunoreactivities in the chicken pretectum. AB - The diversity of nicotinic ACh receptor (AChR) expression in the chick lateral spiriform nucleus (SpL) was assessed using subunit-specific monoclonal antibodies (mAbs) and laser scanning confocal microscopy. The late embryonic SpL was immunoreactive for mAbs against the alpha 2, alpha 5, alpha 7, alpha 8, and beta 2 AChR subunits. Distinct neuronal cell classes were determined using pair-wise staining of mAbs. Approximately 90% of the neurons in the SpL contained both alpha 5-like immunoreactivity (LI) and beta 2-LI, with no neurons having only one of these subunit-LIs. Approximately 70% of the neurons contained alpha 2-LI. All alpha 2-LI neurons contained alpha 5/beta 2-LI; thus, neurons having alpha 2-LI are a subset of those having alpha 5- and beta 2-LI. Fewer neurons, approximately 20%, contained alpha 7-LI. A subset of alpha 7-positive neurons were immunoreactive for other subunits; for example, some alpha 7-positive neurons also contained alpha 2-LI. Fewer than 15% of the neurons contained alpha 8-LI. Some of the alpha 8-LI-containing neurons contained alpha 7-LI. The 14 week post hatch SpL resembles the late embryonic nucleus in the percentage of neurons immunoreactive for alpha 2, alpha 5, alpha 7, alpha 8, and beta 2 AChR subunits, and in the presence of multiple classes based on AChR subunit immunoreactivity. In addition, alpha 4-LI was found in about 20% of the 14 week SpL neurons. Double label immunofluorescence experiments with mAbs to AChRs and to synaptic vesicle antigens showed that most clusters of alpha 5-LI and beta 2-LI are extrasynaptic. The pronounced diversity of AChR subunit expression and the extrasynaptic location of AChR-LI suggest that AChR-like molecules in the SpL do not function solely to respond to transmitter focally released from presynaptic terminals. PMID- 7472458 TI - Repeated treatment with haloperidol and clozapine exerts differential effects on dye coupling between neurons in subregions of striatum and nucleus accumbens. AB - The delayed onset of action of antipsychotic drugs (APDs) during the treatment of schizophrenia has been hypothesized to temporally correlate with the induction of depolarization block in rat mesencephalic dopamine (DA) cell groups. Nevertheless, it is unknown whether these drugs also exert a delayed action on the dopaminoceptive postsynaptic target cells in the striatal complex. Using in vivo intracellular recording and dye labeling techniques, the effects of APDs on dye coupling were examined in subregions of the striatal complex defined by double staining for calbindin immunoreactivity. Rats treated repeatedly with APDs were found to exhibit a 66-71% higher incidence of coupling that occurred in a drug- and a region-specific manner, that is, both drug treatments increased dye coupling in the limbic-associated accumbens shell region whereas only haloperidol increased dye coupling in the motor-related striatal matrix and accumbens core regions. In addition, cells located in regions in which dye coupling was altered also showed significantly higher input resistance. These changes were not observed in response to DA receptor blockade by acute drug administration or when haloperidol was administered for a period sufficient to induce DA receptor supersensitivity but not DA cell depolarization block (i.e., 2 weeks). Therefore, alteration in dye coupling appears to be correlated temporally with the induction of DA cell depolarization block. The finding that both APDs exert a common action on neurons in the accumbens shell region is consistent with its identification as the site of therapeutic drug actions, whereas the capacity of haloperidol to also affect cells in the motor-related matrix and core regions correlates with its high propensity to induce extrapyramidal side effects. PMID- 7472459 TI - Development and regulation of dendritic stratification in retinal ganglion cells by glutamate-mediated afferent activity. AB - In the mature retina, the dendrites of retinal ganglion cells (RGCs) are segregated into either ON or OFF sublaminae of the inner plexiform layer (IPL), but early in development the dendritic processes of these cells are multistratified, ramifying throughout the IPL. We examined the time course of dendritic stratification in developing beta cells, the largest class of ganglion cells in the cat retina, by retrograde labeling of fixed tissue with Dil. Dendritic stratification begins in the central and peripheral retina by embryonic day 50, about 2 weeks before birth and is not fully completed until 5 months postnatally. A clear central-to-peripheral gradient in the incidence of stratified beta cells first becomes evident shortly after birth. This stratification process was effectively halted by short-term intraocular injections (4-11 d) of the glutamate analog 2-amino-4-phosphonobutyrate (APB), which hyperpolarizes rod bipolar cells and ON cone bipolar cells, thereby preventing the release of glutamate by these interneurons. APB treatment did not alter the somal sizes or the tangential extent of the dendrites of developing beta cells, nor did it cause abnormal loss of these neurons. The organization of the inner nuclear layer, containing the APB-sensitive bipolar cells, was also not compromised by such injections. When APB treatment was discontinued there was a rapid resumption of dendritic stratification resulting in a normal incidence of stratified RGCs. Thus, short-term APB treatment causes a delay rather than a permanent arrest of the stratification process.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7472460 TI - Differential expression of AMPA receptor subunits in NOS-positive neurons of cortex, striatum, and hippocampus. AB - AMPA receptor (AMPAR) subunits expression was studied in nitric oxide synthase (NOS)-positive neurons of the adult rat cortex, striatum, and hippocampus, by a double-labeling approach, combining nonradioactive in situ hybridization and immunocytochemistry. The majority of cortical and hippocampal NOS-immunopositive neurons were characterized by a predominant expression of GluR-A and -D mRNA and low or undetectable expression of GluR-B and -C mRNA. In the striatum, the expression profile of AMPAR subunits in NOS-positive neurons differed from that in the other two regions. This is reflected in the overall low expression of all AMPA receptor subunits and the paucity of GluR-D subunit expression that contrasts with the high expression of this subunit in NOS-positive cells in the hippocampus. Double-labeling experiments revealed a substantial correspondence between mRNA and protein levels of AMPAR subunits. Further evidence for the regional diversity of NOS-positive neurons is derived from the expression analysis of glutamate decarboxylase (GAD)-65 and -67 mRNAs. NOS-positive neurons expressed high levels of GAD-65, but not -67 in the cortex, high levels of both forms in the hippocampus, and low or undetectable levels of both mRNAs in the striatum. Despite of these differences, NOS-positive neurons share the common feature of low GluR-B subunit expression, suggesting the presence of AMPAR channels with high Ca2+ permeability, regardless of the regional location. The relative resistance of NOS-positive interneurons in neurodegenerative diseases suggests that glutamate receptor-mediated Ca2+ influx alone does not suffice to explain neuronal vulnerability, and additional factors have thus to be considered. PMID- 7472461 TI - Selective inhibition of axon outgrowth by antibodies to NGF in a model of temporal lobe epilepsy. AB - The molecular basis of the axonal reorganization that follows seizure-induced brain injury is unknown. Elevations in neurotrophins following seizures suggest that growth factors may play a role in this process. After pilocarpine-induced seizures, robust axonal sprouting from dentate granule cells and cholinergic forebrain neurons was evident in the inner molecular layer of the dentate gyrus. Intraventricular infusions of an NGF-specific antibody that blocks NGF biological activity in vitro attenuated the cholinergic axonal sprouting and the increases in cell body size of basal forebrain cholinergic neurons that followed seizure induced injury in vivo. In contrast to its effects on the cholinergic network, the NGF antibody did not decrease the sprouting of dentate granule cell axons into the inner molecular layer. These results suggest that NGF may have a functional and system-specific role in the remodeling of networks that follows repetitive seizures. PMID- 7472462 TI - An obligate role for oxygen in the early stages of glutamate-induced, delayed neuronal death. AB - In vitro models of hypoxic/hypoglycemic injury reveal common mechanisms with glutamate excitotoxicity, but glutamate-induced toxicity in the absence of oxygen has never been directly addressed. Therefore, we assessed neuronal survival and intracellular calcium concentrations ([Ca2+]i) in neonatal hippocampal cultures in response to glutamate in the presence and absence of oxygen. Twenty-four hours of hypoxia alone killed 40% of the initial population, attributable to glutamate receptor-stimulated osmotic lysis. A 5 min glutamate exposure in ambient air killed 80% of the initial population by 24 hr later. When cultures were deprived of oxygen during and for 2-24 hr after excitotoxin exposure, glutamate did not cause additional neuronal death beyond that induced by oxygen depletion alone. Toxicities caused by activation of NMDA, AMPA, or kainate receptors were each ameliorated by oxygen depletion. In the absence of oxygen, glutamate evoked normal increases in [Ca2+]i, indicating that glutamate receptors functioned normally. The glutamate-induced increases in [Ca2+]i were not toxic in the absence of oxygen. In a similar manner, oxygen-depletion prevented neuronal killing by the calcium ionophore, ionomycin. Neuronal death produced by hydrogen peroxide or iron sulfate was not ameliorated by oxygen removal. These oxidants maximally produced only a slow increase in [Ca2+]i as the plasma membrane permeability increased nonspecifically. Therefore, oxygen-based reactions were an essential component of calcium-mediated, delayed neuronal death. PMID- 7472463 TI - Interactions between location and task affect the spatial and directional firing of hippocampal neurons. AB - When rats forage for randomly dispersed food in a high walled cylinder the firing of their hippocampal "place" cells exhibits little dependence on the direction faced by the rat. On radial arm mazes and similar tasks, place cells are strongly directionally selective within their fields. These tasks differ in several respects, including the visual environment, configuration of the traversable space, motor behavior (e.g., linear and angular velocities), and behavioral context (e.g., presence of specific, consistent goal locations within the environment). The contributions of these factors to spatial and directional tuning of hippocampal neurons was systematically examined in rats performing several tasks in either an enriched or a sparse visual environment, and on different apparati. Place fields were more spatially and directionally selective on a radial maze than on an open, circular platform, regardless of the visual environment. On the platform, fields were more directional when the rat searched for food at fixed locations, in a stereotypic and directed manner, than when the food was scattered randomly. Thus, it seems that place fields are more directional when the animal is planning or following a route between points of special significance. This might be related to the spatial focus of the rat's attention (e.g., a particular reference point). Changing the behavioral task was also accompanied by a change in firing location in about one-third of the cells. Thus, hippocampal neuronal activity appears to encode a complex interaction between locations, their significance and the behaviors the rat is called upon to execute. PMID- 7472464 TI - PDGFs protect hippocampal neurons against energy deprivation and oxidative injury: evidence for induction of antioxidant pathways. AB - Platelet-derived growth factors (PDGFs) and PDGF receptors are expressed in brain, where their functions are largely unknown. We tested the hypothesis that PDGFs play a role in promoting the survival of neurons exposed to metabolic and oxidative insults. Exposure of rat and mouse hippocampal cell cultures to glucose deficient medium or the hydroxyl radical-promoting agent FeSO4 resulted in progressive neuronal loss. Pretreatment of cultures with PDGF-AA or PDGF-BB resulted in highly significant attenuation of glucose deprivation- and FeSO4 induced neuronal degeneration. In each injury paradigm the neuroprotective actions of the PDGFs were concentration dependent (3-100 ng/ml). In the case of glucose deprivation, significant protection was seen when cells were exposed to PDGFs prior to, or up to 8 hr following, the onset of glucose deprivation. Pretreatment with PDGFs was required for protection against FeSO4-induced oxidative injury. Western blot and immunocytochemical analyses demonstrated that cultured embryonic hippocampal neurons expressed both PDGF alpha- and beta receptors. PDGFs induced tyrosine phosphorylation of several proteins including a band at 180 kDa, the molecular weight of PDGF receptors. Induction of peroxide accumulation in neurons by FeSO4 was attenuated in cultures pretreated with PDGFs, suggesting that PDGFs enhanced cellular antioxidant mechanisms. Measurements of anti-oxidant enzyme activities in control and PDGF-treated cultures showed that both PDGF-AA and PDGF-BB increased both catalase and glutathione peroxidase activities, and PDGF-AA also increased superoxide dismutase activities. These findings suggest that PDGFs, which are widely expressed in brain and induced in response to injury, may play roles in protecting neurons against metabolic and oxidative insults. PMID- 7472465 TI - Cholinergic, GABAergic, and glutamate-enriched inputs from the mesopontine tegmentum to the subthalamic nucleus in the rat. AB - In order to clarify the origin and to examine the neurochemistry and synaptology of the projection from the mesopontine tegmentum (MTg) to the subthalamic nucleus (STN), rats received discrete deposits of anterograde tracers in different regions of the MTg. Anterogradely labeled fibers were examined in the light and electron microscopes. The distribution of GABA or glutamate immunoreactivity was examined by post-embedding immunocytochemistry. The anterograde tracing demonstrated that the projection to the STN arises from at least three divisions of the MTg: the area defined by the cholinergic neurons of the pedunculopontine region (PPN-Ch 5), the more medial and largely noncholinergic midbrain extrapyramidal area (MEA) and to a lesser extent the laterodorsal tegmental nucleus (LDTg). Post-embedding immunocytochemistry revealed that there are GABA immunopositive and immunonegative components to this projection and at least a proportion of the GABA-immunonegative component is enriched in glutamate immunoreactivity. The similarity of the morphology, trajectory and synaptology of the anterogradely labeled fibers and the choline acetyltransferase (ChAT) immunopositive fibers supports the proposal that at least part of the projection is cholinergic. The terminals anterogradely labeled from the MTg and the ChAT immunoreactive terminals form asymmetrical synapses with the dendrites and spines of subthalamic neurons. Both anterogradely labeled and ChAT-positive terminals make convergent synaptic contacts with GABA-immunoreactive terminals that form symmetrical synaptic contacts and are probably derived from the globus pallidus. Taken together these findings imply that the MTg sends cholinergic, GABAergic and glutamatergic projections to the STN where at least one of the functional roles is to modulate the indirect pathway of information flow through the basal ganglia that is carried via the pallidosubthalamic projection. PMID- 7472466 TI - Purinergic receptors and their activation by endogenous purines at perisynaptic glial cells of the frog neuromuscular junction. AB - Glial cells are closely associated with synapses and are modulated by neurotransmitters released during synaptic transmission. At many synapses, ATP is released during synaptic transmission and is involved in cell-cell signaling. Since glial cells have purinoceptors, it is possible that ATP mediates synaptic neuron-glia signaling. This work aims at determining which types of purinoceptors are present on perisynaptic Schwann cells, the perisynaptic glial cells at the frog neuromuscular junction, and test their sensitivity to endogenous purines by monitoring the relative changes of intracellular Ca2+. Local application of ATP induced the release of Ca2+ from internal stores. Adenosine induced Ca2+ responses that were blocked by A1 receptor antagonists and mimicked by an A1 receptor agonist and were caused by the release of Ca2+ from internal stores via a pertussis toxin-sensitive G-protein. A2 receptor antagonists had no effect on Ca2+ responses induced by adenosine. Me-S-ATP, an ATP analog, triggered Ca2+ release from internal stores via a pertussis toxin-sensitive G-protein, consistent with the activation of P2Y receptors. L-AMP-PCP, another ATP analog, induced Ca2+ entry mainly through L-type Ca2+ channels by a pertussis toxin insensitive mechanism, consistent with the activation of P2X receptors. Blockade of adenosine receptors did not affect glial Ca2+ responses induced by nerve evoked transmitter release. However, blockade of ATP receptors reduced the size and increased the delay of the responses. Hence, purinoceptors are present on the perisynaptic Schwann cells and are activated by endogenous ATP released during synaptic transmission. PMID- 7472467 TI - Dynamics of central nervous 5-HT1A-receptors under psychosocial stress. AB - It is well established that stress leads to changes in the serotonergic system. In order to gain a better understanding of the effects of recurrent stressful experiences on the serotonergic system, changes in the 5-HT1A-receptor system resulting from different periods of psychosocial stress (PSS) were analyzed in the present study. Male tree shrews (Tupaia belangeri) were submitted to subordination stress for 2, 10, 21, and 28 d. 5-HT1A-receptor binding was quantified by in vitro receptor autoradiography using the agonist 3H-8-OH-DPAT (3H-8-hydroxy-2-(di-n-propylamino)tetralin. PSS caused a downregulation of 3H-8 OH-DPAT binding sites in cortical areas and in the hippocampus. After 10, 21, and 28 d of PSS, the number of binding sites was reduced in layers V and VI of the posterior cingulate cortex (by 34%). After 28 d of PSS, the number of binding sites was reduced in the parietal cortex (by 18%), in the prefrontal cortex (by 16%), in the regio retrobulbaris (by 8%), and in region CA1 of the hippocampus (by 11%). In the raphe nuclei, no PSS-induced downregulation of 5-HT1A-receptors occurred. A transient increase in 3H-8-OH-DPAT binding was observed in the claustrum after 2 d of PSS (by 15%). There were also transient decreases in affinities for the radioligand probably representing receptor desensitization, for example, in the dorsal raphe nucleus after 28 d of PSS. In conclusion, the dynamic 5-HT1A-receptor changes occurring during PSS include downregulation and transient desensitization of receptors. They reflect regulatory mechanisms which probably lead to destabilization of the serotonergic system during prolonged PSS. PMID- 7472468 TI - Basic fibroblast growth factor increases division and delays differentiation of dopamine precursors in vitro. AB - Growth factors have been shown to be involved in the regulation of division and differentiation of neuroepithelial cells. In the present study we examined the ability of various factors to influence the development of dopamine precursors. Primary neuronal cultures were prepared from the embryonic day 12 (E12) rat ventral mesencephalon, a time and place which coincides with the beginning of the birth of the dopamine neurons of the substantia nigra pars compacta. At low plating density in serum-free medium, the dopamine precursors divide for approximately 1 d in vitro. We report here that basic fibroblast growth factor (bFGF) can expand the period of dopamine precursor division at least until day 8 in culture, which is well beyond the normal division of these cells. This increase in cell division was accompanied by a delay in differentiation as compared to untreated control cultures. Upon differentiation, the high-affinity dopamine uptake values in bFGF-treated cultures were 20 times maximal control values. Mature dopamine neurons appeared at the same time as astrocytes, which may be playing a role in inducing dopamine neuron differentiation. IGF-I, GDNF, and EGF were unable to mimic the effect of bFGF on division and differentiation of dopamine precursors. Expanding in vitro the number of dopamine precursors provides tissue that may be suitable for transplantation in patients with Parkinson's disease. PMID- 7472469 TI - Local modulation of hippocampal acetylcholine release by dopamine D1 receptors: a combined receptor autoradiography and in vivo dialysis study. AB - The modulation of in vivo hippocampal ACh release by dopaminergic D1 and D2 receptors was examined in this study. Additionally, in an attempt to ascertain the location of these receptors in relation to hippocampal cholinergic terminals, fimbriaectomy and quantitative autoradiography were used. Following unilateral fimbriaectomy, whereby at least 50% of hippocampal cholineacetyltransferase (ChAT) activity was lost, a significant ipsilateral decrease in D1/3H SCH23390 binding was observed in the molecular layer of the dentate gyrus while hippocampal D2/3H raclopride binding was unaffected. The effects of prototypical D1 and D2 receptor agonists and antagonists on hippocampal ACh release were examined next using in vivo dialysis in freely moving rats. The D1 agonist SKF 38393 (10 microM to 100 microM) administered directly into the hippocampus via the dialysis probe stimulated ACh release in a concentration dependent manner. The effect of the agonist was blocked by the coadministration of the D1 receptor antagonist SCH 23390 (1 microM), which by itself failed to modulate ACh release. In contrast, neither the D2 agonist quinpirole (1-10 microM) nor the D2 antagonist sulpiride (1-10 microM) had any direct effect on hippocampal ACh release. Additionally, the infusion of these D1 and D2 drugs in the septal area failed to affect hippocampal ACh release. Taken together, these results suggest that a proportion of hippocampal D1 receptors are located on cholinergic nerve terminals and that dopamine, acting via D1 receptors, can locally stimulate hippocampal ACh release. PMID- 7472470 TI - Intraneuronal delivery of protein kinase C pseudosubstrate leads to growth cone collapse. AB - Axonal navigation during development requires that cues present in the extracellular environment be capable of modifying the structure of the cone in a dynamic way. Protein kinase C (PKC) has long been suspected to be one of the multiple molecular relays present in the terminal structure of the developing axon and involved in the transduction of extracellular signals. The latter proposal is, however, based on the use of drugs or of protocols leading to pleiotropic and often nonspecific effects. In the present study, we have taken advantage of the discovery of a peptide capable of translocating across biological membranes and to accumulate in the cytoplasm and nucleus of cells in culture, to internalize a highly specific peptidic inhibitor of PKC. We demonstrate that linking the two peptides (vector and PKC inhibitor) allows the internalization of the latter in live cells, specifically inhibits PKC and provokes a rapid modification of growth cone morphology. This set of data thus establishes that a peptidic inhibitor of PKC activity, once internalized, provokes a change in growth cone morphology, reminiscent of the collapse phenotype. In addition, the present study describes a new efficient and harmless way to introduce pharmacologically active substances in neural cells in culture. PMID- 7472471 TI - Presynaptic glutamate levels in tonic and phasic motor axons correlate with properties of synaptic release. AB - Synaptic glutamate release involves the accumulation of cytoplasmic glutamate in synaptic vesicles, whereafter it is released by triggered exocytosis. As glutamatergic terminals are known to be functionally diverse it was of interest to examine whether the presynaptic glutamate supply differs between individual axon terminals with distinct release properties. The glutamatergic terminals in the crustacean neuromuscular system system comprise a "phasic" type which shows fatigue of release during repetitive stimulation, and a "tonic" type which can maintain transmission for long periods. Quantitative immunogold analysis showed that the axons in a tonic nerve innervating slow muscles in the abdomen contained two times higher levels of glutamate labeling over axoplasmic matrix and over mitochondria, as compared to the corresponding elements in a phasic nerve. Similar results were obtained when adjacent phasic and tonic axons in a mixed nerve innervating leg muscles were compared. In the terminal regions of tonic and phasic axons the glutamate labeling differed correspondingly over axoplasmic matrix and mitochondria, while the synaptic vesicles showed a similar strong accumulation of labeling in both types of terminal. The level of labeling for glutamine, a glutamate precursor, was closely similar in phasic and tonic axons. The axoplasmic glutamate concentration was estimated to be in the low millimolar range, through comparison with coprocessed conjugates with known glutamate concentration. These results show that fatigue-resistant tonic axons and terminals contain higher levels of glutamate than fatiguable phasic axons, presumably representing an adaptation to the markedly different impulse activities in the two types of neuron. The axonal glutamate concentrations are in the range of the Km value for vesicular glutamate transport. Thus in tonic axons the high glutamate level appears to promote an efficient refilling of synaptic vesicles during sustained release, while in phasic axons the refilling should be slower which is compatible with an infrequent release. PMID- 7472472 TI - Stress-induced sensitization and glucocorticoids. I. Sensitization of dopamine dependent locomotor effects of amphetamine and morphine depends on stress-induced corticosterone secretion. AB - Repeated exposures to stress sensitize motor and addictive effects of drugs of abuse. Recently, it has been shown that stress-induced behavioral sensitization depends on the secretion of glucocorticoids. We investigated if sensitization of dopamine-dependent effects of psychostimulants and opioids was influenced by glucocorticoid. Sensitization of the dopaminergic response to drugs is considered the neural substrate of behavioral sensitization and has been implicated in vulnerability to drug abuse. Dopamine-dependent effects of psychostimulants and opioids were evaluated by injecting either amphetamine into the nucleus accumbens (10 micrograms/side) or morphine into the ventral tegmental area (VTA) (1 microgram/side). The locomotor response to psychostimulants and opioids injected in these brain areas depends on the mesencephalic dopaminergic transmission. Drug induced locomotion was compared in male rats in which corticosterone secretion was either in +tct or experimentally suppressed by an adrenalectomy associated with a substitutive treatment reproducing basal levels of the hormone. Eight days of food restriction (80% of the initial body weight) were used as a stressor. Suppression of stress-induced corticosterone secretion abolished food restriction induced sensitization of the locomotor effects of intra-accumbens amphetamine and intra-VTA morphine. This effect was corticosterone dependent since the restoration of corticosterone levels in the range of those induced by stress totally reinstates sensitization. Our results suggest that glucocorticoids control stress-induced sensitization by changing the sensitivity of the mesencephalic dopaminergic transmission to drugs of abuse. Since dopaminergic effects of drugs are related to their addictive properties, secretion of glucocorticoids may be one of the factors determining the enhanced vulnerability to drugs observed in stressed subjects. PMID- 7472473 TI - Stress-induced sensitization and glucocorticoids. II. Sensitization of the increase in extracellular dopamine induced by cocaine depends on stress-induced corticosterone secretion. AB - Secretion of glucocorticoids seems to control stress-induced sensitization of the behavioral effects of drugs of abuse by acting on the mesencephalic dopaminergic transmission, the principal neural substrate of sensitization. In order to investigate the mechanisms of this interaction between glucocorticoids and dopamine, we studied the sensitization of the increase in extracellular concentration of dopamine induced by cocaine in male rats in which corticosterone secretion was either intact or blocked. Extracellular concentrations of dopamine were evaluated in the nucleus accumbens of freely moving animals by means of microdialysis. Metyrapone, an inhibitor of corticosterone synthesis, was used to block stress-induced corticosterone secretion. Food-restriction (90% of the initial body weight) was the stressor used to induce sensitization. It was found that metyrapone (100 mg/kg s.c. twice a day for 8 d) suppressed stress-induced sensitization of the increase in accumbens dopamine induced by cocaine (10 mg/kg, i.p.) and sensitization of cocaine-induced locomotion Metyrapone suppressed both the development and the expression of sensitization. Thus, sensitization was equally blocked when the metyrapone treatment started either 1 d before the start of food-restriction or 8 d later, that is, when food-restriction-induced sensitization to cocaine was already established. In conclusion, our results suggest that glucocorticoids modify sensitization of the behavioral effects of cocaine by acting on extracellular concentrations of dopamine. Since addictive properties of psychostimulants seem mediated by the increase in extracellular concentrations of dopamine they induce, these findings may have implications for the development of new therapeutic strategies of addiction. PMID- 7472474 TI - Cyclic-GMP enhances light-induced excitation and induces membrane currents in Drosophila retinal photoreceptors. AB - Phototransduction in the Drosophila retina appears to require the phosphoinositide signaling cascade following receptor/G-protein activation. Subsequent opening of membrane cationic channels causes excitation. The biochemical events underlying channel opening and regulation of sensitivity remain largely unknown. Evidence is mounting that phototransduction in Drosophila and other invertebrate species may additionally involve the second messenger, cyclic-GMP (cGMP). We report that exogenous cGMP influenced Drosophila retinal phototransduction in two ways. In whole cell tight-seal voltage-clamp experiments, membrane permeant cGMP analog, 8-bromo-cyclic-GMP (8-Br-cGMP), induced membrane currents and dramatically enhanced light-induced currents. The currents induced by 8-Br-cGMP possessed reversal potentials similar to those induced by light. The magnitudes of cGMP-induced currents exhibited marked dependence on intensity of background illumination. Potential direct or modulatory roles of cGMP in Drosophila phototransduction are discussed. PMID- 7472475 TI - Differential processing of semicircular canal signals in the vestibulo-ocular reflex. AB - Selective semicircular canal inactivation and three-dimensional eye movement recordings have been used to investigate the spatial organization of vestibular signals in the vestibulo-ocular reflex (VOR) of rhesus monkeys. In animals with one pair of semicircular canals inactivated, afferent activity no longer codes all spatial components of head angular velocity. if it were the activation pattern of semicircular canal afferents alone that determines VOR slow phase eye velocity, the head velocity components along the sensitivity vectors of the remaining intact semicircular canals would determine the orientation of slow phase eye velocity. Thus, angular head velocity and slow phase eye velocity would not necessarily always align. Alternatively, if vestibulo-ocular signals coded absolute angular head motion in space based on both semicircular canal and otolith afferent information, one might expect a spatial transformation of the encoded head angular velocity signals such that slow phase eye velocity and angular head velocity continue to spatially align. Examination of the VOR at different frequencies between 0.01 Hz and 1 Hz revealed a frequency-specific spatial organization of vestibulo-ocular signals. Mid and high frequency vestibulo-ocular responses were determined exclusively by the orientation of the sensitivity vectors of the remaining intact semicircular canals. In contrasts, low frequency vestibulo-ocular responses were largely determined by the orientation of the head relative to gravity. These low frequency responses after selective semicircular canal inactivation could be predicted and simulated by a simple model where semicircular canal signals are spatially transformed from a head-fixed to a space-fixed (inertial) representation of angular head velocity. These findings suggest that low frequency vestibulo-ocular responses are dominated by inertial vestibular signals that detect absolute head motion in space based on both semicircular canal and otolith afferent information. Inertial vestibular signals are likely to contribute to head control and motor coordination of gaze, head and body posture. PMID- 7472476 TI - Serotonin reduces inhibition via 5-HT1A receptors in area CA1 of rat hippocampal slices in vitro. AB - We studied the effects of serotonin (5-HT) on intrinsic and synaptic responses of hippocampal CA1 cells. The effects were partially mimicked by the 5-HT1A receptor agonist, 8-OH-DPAT, and prevented by the 5-HT1A receptor antagonist, NAN-190. Polysynaptic fast and slow inhibitory postsynaptic potentials (IPSPs) were reduced in amplitude by 60-70% following application of both 5-HT and 8-OH-DPAT. Monosynaptic fast IPSPs were reduced by 60% and slow IPSPs by 90% following application of both drugs. Since there is a temporal overlap of fast and slow IPSPs, the reduction in fast IPSPs could have arisen indirectly from the larger effect of 5-HT on slow IPSPs. To overcome this problem we blocked the slow IPSPs with new, potent GABA-B antagonists, but still observed a similar reduction in the fast IPSP with 5-HT and 8-OH-DPAT. However, the reductions in the fast IPSPs could also have arisen from the 5-HT-induced total conductance increases. Using single-electrode voltage clamp and intracellular K+ channel blockers we still observed similar changes. 5-HT and 8-OH-DPAT had no effect upon GABA-A-mediated currents evoked by iontophoretic GABA application to the dendrites or the soma of CA1 pyramidal cells, Putative inhibitory internuerons were hyperpolarized by 5-HT and their evoked EPSPs strongly reduced by 5-HT and 8-OH-DPAT. Our data indicate that 5-HT modulates fast and slow synaptic inhibition of principal cells using presynaptic mechanisms involving the inhibition of inhibitory interneurons. PMID- 7472477 TI - Impaired peripheral nerve regeneration in a mutant strain of mice (Enr) with a Schwann cell defect. AB - Schwann cell-axon interactions in the development, maintenance, and regeneration of the normal peripheral nervous system are complex. A previously described transgene-induced insertional mutation (BPFD#36), now referred to as Enervated (Enr), results in disrupted Schwann cell-axon interactions. In this report, after a crush or transection injury to Enr peripheral nerves, we demonstrate impaired nerve regeneration. There are fewer myelinated fibers per mm2 and thinner myelin sheaths surrounding regenerating axons in the nerves of homozygous mutant mice compared to wild type mice at 28 d after crush injury to the sciatic nerve. Abnormal Schwann cell-axon interactions remain in Enr/Enr animals as evidenced by the relatively frequent ultrastructural finding of unmyelinated large diameter axons in the regenerating nerves. Additionally, nerve graft experiments indicate that the impairment in regeneration is due to a Schwann cell defect. Morphologic and morphometric findings in conjunction with molecular analysis of regenerating nerves suggest that the Enr defect causes a disruption in the ability of "early" Schwann cells to differentiate to a more mature phenotype. In mutant homozygous and wild type nerves at 7 d after crush injury there are similar levels of mRNA for the low-affinity nerve growth factor receptor, but in the mutant homozygous regenerating nerves there is 11-fold less mRNA for glial fibrillary acidic protein, a more mature phenotypic marker of Schwann cells. This Schwann cell differentiation defect likely accounts for both the peripheral neuropathy and impaired nerve regeneration observed in Enr mice. PMID- 7472478 TI - Early patterns of migration, morphogenesis, and intermediate filament expression of subventricular zone cells in the postnatal rat forebrain. AB - Recent studies using retroviral labeling of subventricular zone (SVZ) progenitors in vivo in neonatal rats have directly demonstrated the generation of both astrocytes and oligodendrocytes from these progenitors. In the present study, we used a recombinant retroviral vector encoding beta-galactosidase, and analyzed brains within the first week after retroviral injection to trace the early routes that SVZ cells take as they migrate into white matter and cortex and characterized the early morphological and antigenic changes that accompanied their differentiation. SVZ cells follow specifically definable migratory routes as they colonize the cortex and subcortical white matter. Glial progenitors do not populate the cortex in a systematic, laminar fashion, as do neuroblasts. The abundance of labeled progenitors in radial arrangements and the close apposition of many immature cells to vimentin+ radial glial processes, suggest that glial progenitors migrate along radial glia. Labeled SVZ cells, which displayed a simple, unipolar or bipolar morphology, lacked detectable vimentin and nestin intermediate filaments. Similarly, beta-galactosidase-positive cells in white matter lacked these filaments. In contrast, labeled, multipolar cells in the cortex, and a few of the immature-appearing cortical cells expressed nestin and vimentin. At these early time points, GFAP was not detected in beta-galactosidase labeled cells. Multipolar cells in cortex frequently displayed processes extending toward and contacting blood vessels. These observations suggest that the expression of nestin and vimentin occurs after progenitors emigrate from the SVZ and that filament expression and contact with blood vessels represent an early stage of astrocyte differentiation. PMID- 7472479 TI - Delayed postischemic hypothermia: a six month survival study using behavioral and histological assessments of neuroprotection. AB - In the gerbil, brief global forebrain ischemia induces profound habituation and working memory impairments that stem from delayed hippocampal CA1 death. Short duration postischemic hypothermia has been shown to reduce CA1 loss, but such reports are controversial, as it is thought that protection may be transient. The purpose of this study was to investigate whether prolonged postischemic hypothermia provided long-term CA1 and functional neuroprotection. Previously, 90% of anterior CA1 neurons were rescued (30 d survival) when 24 hr of hypothermia (32 degrees C) was induced 1 hr following a 5 min occlusion that otherwise produced more than 95% loss (Colbourne and Corbett, 1994). We now find about 70% CA1 savings with this same hypothermic treatment in gerbils that survived for 6 months postischemia. While this is a significant reduction from 30 day survival (medial CA1 only), it nonetheless shows, for the first time, persistent, if not permanent neuroprotection, especially in middle and lateral CA1. In addition, in non-treated animals, ischemia impaired learning in an open field and T-maze for up to 6 months. Postischemic hypothermia significantly reduced these deficits. Hypothermia (32 degrees), when initiated 4 hr after ischemia, rescued approximately 12% of CA1 neurons at 6 months with a slight behavioral benefit. Milder hypothermia (34 degrees C, 1-25 hr postischemia, 30 d survival) also reduced habituation impairments and saved approximately 60% of CA1 neurons. Similar trends were found at more caudal CA1 levels. These results clearly show that postischemic hypothermia provides effective and long-lasting neuroprotection, which depends upon the delay to initiation, duration, and degree of cooling and survival time. The protracted functional and histological benefit observed justifies further basic and clinical investigation. PMID- 7472480 TI - Integration of chemosensory and hormonal cues is essential for mating in the male Syrian hamster. AB - Mating behavior in the male hamster is dependent upon both chemosensory and hormonal cues, and copulation is abolished if either signal is interrupted. Through reciprocal interactions of these signals, chemosensory stimuli increase circulating testosterone in the male, and the male's hormonal status influences his attraction to female pheromones. Furthermore, anatomical data suggest that these signals are transmitted through parallel pathways in separate subdivisions of the medial amygdaloid nucleus, bed nucleus of the stria terminalis (BNST) and medial preoptic area (MPOA). The aim of this study was to determine if the integration of chemosensory and hormonal cues is essential for mating. We combined an intracerebral implant of testosterone in BNST/MPOA with removal of a single olfactory bulb (UBx), ipsilateral or contralateral to the steroid implant. Previous studies have demonstrated that testosterone implants which stimulate androgen receptor-containing neurons in posteromedial BNST and MPOA can increase mounting in males castrated for 12 weeks. Moreover, unilateral bulbectomy alone does not prevent mating. In the present study, ipsilateral UBx prevents communication between hormonal and chemosensory circuits. Sexually experienced males were used. Twelve weeks after castration, a single olfactory bulb was removed, and each male received a testosterone-filled cannula (23 ga) directed at the MPOA. Two weeks later, sexual activity increased in six males with implants in BNST/MPOA and contralateral UBx, but copulation was not restored in eight males with ipsilateral UBx despite equivalent implant placement. This study demonstrates that communication between neurons receiving hormonal signals and chemosensory cues is required for mating behavior. PMID- 7472481 TI - A comparison of the effects of medial prefrontal, cingulate cortex, and cingulum bundle lesions on tests of spatial memory: evidence of a double dissociation between frontal and cingulum bundle contributions. AB - Rats were trained on an automated delayed nonmatching-to-position (DNMP) task. They then received cytotoxic lesions in either the medial prefrontal cortex (n = 13) or the cingulate and retrosplenial cortices (n = 8), or radiofrequency lesions in either the fornix (n = 6) or the cingulum bundle (n = 8). Twelve animals served as surgical controls. Only the fornical and medial prefrontal lesions disrupted DNMP performance, both groups showing a loss of accuracy and an increase in bias. The rats were then trained on a lever discrimination and reversal task, the medial prefrontal and fornical groups showing evidence of an increase in bias when compared with the cingulate cortex group. Finally, the rats were trained on a forced alternation task in a T-maze. Marked deficits were observed in the fornix and cingulum bundle groups, but the medial prefrontal and cingulate groups were unimpaired. The double dissociation between the effects of the prefrontal and cingulum bundle lesions highlights the very different nature of the two spatial tasks (DNMP and T-maze alternation), even though both involved a nonmatching rule. These findings may reflect the involvement of divergent outputs from the fornix-anterior thalamic pathway. One possibility is that anterior thalamic projections to the medial prefrontal cortex are concerned with processing egocentric information, while anterior thalamic projections to temporal regions via the cingulum bundle are concerned with allocentric information. The results also indicate that the effects of conventional lesions in the cingulate cortex and medial prefrontal cortex may be compromised by additional damage to the cingulum bundle. PMID- 7472482 TI - Age-associated impairments in a test of attention: evidence for involvement of cholinergic systems. AB - We trained three groups of rats, young (Y; 3 months at the start of study), middle aged (MA; 15 months), and aged (AG; 22 months), in the serial five-choice serial reaction time task, a test of attention. There were clear age-related differences in task acquisition: Y acquired the task quicker than MA rats, which learned faster than AG rats. A subgroup of AG rats [AG(I)] could not reach criterion (> 80% correct, < 20% omissions under standard conditions of 0.5 sec stimulus duration, 5 sec limited hold). Accordingly, they were tested under conditions of 1 sec stimulus duration. Having acquired the task, under standard conditions both MA and AG groups were slower to make a correct response but not to collect the food reward. Furthermore, parameter changes, particularly reductions in stimulus duration and intensity, revealed further age-related changes in accuracy. Following completion of these studies, animals were trained in a simpler one-choice task. Importantly, reducing stimulus duration/intensity in this task revealed no differences between Y and MA/AG groups, although AG(I) rats were impaired. This dissociation between MA/AG impairments in the one- and five-choice task suggests that these animals may show attentional deficits compared with Y rats, which are independent of changes in sensory (visual), motor function, or motivation. Finally, the MA deficit in attention was partially reversed by tacrine pretreatment (3 mg/kg). Also scopolamine (0.01-0.075 mg/kg) and mecamylamine (0.3-5 mg/kg) pretreatment impaired choice accuracy of MA but not Y rats. Taken together, the drug studies imply that the attentional deficits may at least be partially due to changes in cholinergic function. PMID- 7472483 TI - Differential susceptibility of human CNS-derived cell populations to TNF dependent and independent immune-mediated injury. AB - We examined whether oligodendrocytes, neurons, and astroglia derived from the human central nervous system differ in susceptibility to injury mediated by tumor necrosis factor (TNF)-alpha and by activated CD4+ T cells acting via a TNF independent mechanism. Injury was assessed either as cell membrane-directed (lysis), measured by 51chromium (Cr) or lactate dehydrogenase (LDH) release, or nucleus-directed (apoptosis), measured by morphologic features based on propidium iodide (PI) staining and by DNA fragmentation measured by a terminal transferase (TdT)-mediated dUTP biotin nick end labeling technique (TUNEL). TNF did not induce 51Cr or LDH release in any cell targets, but did induce nuclear (66 +/- 2% of cells) and DNA (68 +/- 2% of cells) fragmentation selectively in the oligodendrocytes over 96 hr. At this time, there was no significant loss of oligodendrocyte cell number. Nuclear injury could be induced in neurons by serum deprivation and in malignant astrocytes by the combination of TNF and low serum. CD4+ T cells activated with phytohemagglutin (pha) or anti-CD3 plus interleukin-2 induced significant 51Cr and LDH release in all target cells tested; only pha activated CD4+ T-cell cocultures showed reduced target cell numbers. Significant nuclear fragmentation was observed only for glioma cells (22 +/- 1% of cells). Differences in susceptibility to different immune effector mechanisms and in the nature of the injury response to the same effector mediator among human CNS derived neural cells will need to be considered in design of therapeutic strategies aimed at protecting or limiting target cell injury consequent to disease or trauma. PMID- 7472484 TI - Disruption of cerebellar maturation by an antimitotic agent impairs the ontogeny of eyeblink conditioning in rats. AB - This study represents an attempt to establish a relationship between maturation of the cerebellum and the ontogeny of eyeblink conditioning in the rat. Experiments 1 and 2 examined the effects of disrupting cerebellar maturation by neonatal exposure to the antimitotic agent methylazoxymethanol (MAM) on the ontogeny of eyeblink conditioning in infant rats. Experiment 1 demonstrated that neonatal exposure to MAM on Postnatal Day 4 (PND4) and 7 severely disrupted cerebellar maturation. This effect appeared to be specific in that there was no overt dysmorphology in other brain regions. MAM treatment also severely disrupted associative eyeblink conditioning in rats given training on PND24 and 25. However, exposure to MAM had no effect on the unconditioned response, T-maze delayed alternation, or conditioned suppression of ongoing behavior. In Experiment 2, MAM was given on PND4 and 7 and pups were tested behaviorally on PND17-18, 20-21, or 31-32. Cerebellar hypoplasia was most dramatic shortly after exposure. The cerebellar cortex continued to mature after exposure to MAM, but development of morphological endpoints examined here were static from PND19 to 33. Eyeblink conditioning was impaired at all ages, indicating that there was no functional recovery following neonatal exposure to MAM over the age range tested. These experiments suggest that normal cerebellar maturation may be important for the ontogeny of eyeblink conditioning. PMID- 7472485 TI - Basal forebrain cholinergic lesions disrupt increments but not decrements in conditioned stimulus processing. AB - Magnocellular neurons in the basal forebrain provide the major cholinergic innervation of cortex. Recent research suggests that this cholinergic system plays an important role in the regulation of attentional processes. The present study examined the ability of rats with selective immunotoxic lesions of these neurons (made with 192 IgG-saporin) to modulate attention within an associative learning framework. Each rat was exposed to conditioned stimuli (CS) that were either consistent or inconsistent predictors of subsequent cues. Intact control rats showed increased CS associability when that cue was an inconsistent predictor of a subsequent cue, whereas lesioned rats were impaired in increasing attention to the CS when its established relation to another cue was modified. In a separate experiment designed to test latent inhibition, it was shown that removal of the corticopetal cholinergic neurons spared a decrement in associability that occurs when rats are extensively preexposed to a CS prior to conditioning. These data indicate that the cholinergic innervation of cortex is critical for incrementing, but not for decrementing attentional processing. The specific behavioral tests used to assess the role of the basal forebrain cholinergic system in the present study were previously used to identify a role for the amygdala central nucleus in attention (Holland and Gallagher, 1993b). Those studies, together with the results in this report, indicate that regulation of attentional processes during associative learning may be mediated by projections from the amygdala to the basal forebrain cholinergic system. PMID- 7472486 TI - Hippocampal lesions disrupt decrements but not increments in conditioned stimulus processing. AB - Studies recording hippocampal neural activity show widespread registration of events during associative learning. Recent computational models of hippocampal function have stressed its role in attentional processes specified by well developed modern theories of associative learning. These modeling efforts are largely aimed at accounting for the behavioral outcomes of damage to the hippocampal system in terms of underlying changes in information processing. Two experiments examined the effects of neurotoxic lesions of the hippocampus on changes in attentional processing of a conditioned stimulus (CS) in appetitive Pavlovian conditioning in rats. In Experiment 1, hippocampal lesions eliminated the reduction in associability of a CS usually produced by preexposure to that cue (latent inhibition). In Experiment 2, hippocampal lesions interfered with the loss in associability of a CS normally produced when that CS consistently predicts another event. In contrast, in Experiment 2, hippocampal lesions did not prevent the enhancement of CS associability when a previously consistent predictive relation between two events was made inconsistent. This research supports previous claims that the hippocampus is involved in regulating the processing of CSs in Pavlovian conditioning, and provides new evidence for a hippocampal role in decremental, but not incremental, changes in attention. PMID- 7472487 TI - Chronic cocaine exposure in the fetal rhesus monkey: consequences for early development of dopamine neurons. AB - The effects of cocaine on the development of neurons containing tyrosine hydroxylase (TH) were investigated in fetal rhesus macaques. Pregnant monkeys were treated with cocaine 3 mg/kg or saline i.m., four times a day from day 18 of pregnancy until days 40 or 60. Cocaine concentration in plasma from females thus treated was approximately 800 ng/ml 10-20 min following injection. At the time of C-section, plasma levels of cocaine in fetal blood were 231 +/- 70 ng/ml. The brains from 40 and 60 d old fetuses were examined using immunocytochemistry and in situ hybridization. The appearance of neurons containing TH by day 40 of gestation was not different between control and cocaine-treated fetal monkeys. In both groups (N = 3 each) TH-IR neurons and TH mRNA were located in the mesencephalon and dorsal hypothalamus by day 40, and fiber projections extended to the developing striatum. Also in the day 60 fetuses, the TH-IR neurons were distributed similarly in both groups (N = 5 each), but the TH mRNA content, measured by quantitative in situ hybridization, was reduced in the substantia nigra (SN) and ventral tegmental area (VTA) after cocaine treatment. These data suggest that exposure to cocaine in fetal life does not affect the development of TH or the expression of its mRNA on day 40 of gestation. By day 60, however, the expression of TH mRNA was significantly reduced. This latter effect can be explained by reduced dopamine synthesis in the cocaine-treated fetuses. PMID- 7472488 TI - In vivo actions of insulin-like growth factor-I (IGF-I) on brain myelination: studies of IGF-I and IGF binding protein-1 (IGFBP-1) transgenic mice. AB - To study the effects and mechanisms of insulin-like growth factor I (IGF-I) on brain myelination in vivo, the morphology of myelinated axons and the expression of myelin specific protein genes have been examined in transgenic (Tg) mice that overexpress IGF-I and that those ectopically express IGF binding protein-1 (IGFBP 1), a protein that inhibits IGF-I actions when present in molar excess. Our data show that the percentage of myelinated axons and the thickness of myelin sheaths are significantly increased in IGF-I Tg and decreased in the IGFBP-1 mice. Cerebral cortical proteolipid protein (PLP) and myelin basic protein (MBP) mRNAs consistently exhibit approximately 200% increases in IGF-I Tg mice and approximately 50% decreases in IGFBP-1 Tg mice. The percentage of oligodendrocytes labeled with a PLP cRNA probe in the corpus callosum and cerebral cortex also is increased in IGF-I Tg mice and reduced in IGFBP-1 Tg mice, suggesting that IGF-I promotes oligodendrocyte survival and/or proliferation. The alterations in the number of oligodendrocytes, however, can not completely account for the changes in myelin gene expression. These results strongly indicate that IGF-I increases myelination by increasing the number of myelinated axons and the thickness of myelin sheaths, the latter by mechanisms that involve stimulation of the expression of myelin protein genes and increase of oligodendrocyte number. PMID- 7472489 TI - A timetable of events during programmed cell death induced by trophic factor withdrawal from neuronal PC12 cells. AB - We describe a timetable of events during programmed cell death (PCD) in neuronal PC12 cells, specifically, Ras signaling, immediate-early gene (IEG) expression, DNA fragmentation and commitment to PCD. Commitment occurs over a period from 10 20 hr after NGF withdrawal. Ras signaling declines rapidly after NGF removal, reaching minimal levels within 2-4 hr, well before the onset of commitment. DNA fragmentation, detected by TUNEL reaction, begins about 24 hr after NGF withdrawal, well after all cells are committed, but coincident with the onset of cell dissolution previously determined by trypan blue exclusion (Mesner et al., 1992). Among the IEGs studied here, c-jun and TIS21 are expressed within 6 hr after NGF withdrawal. Expression of c-fos, egr-1, and TIS11 does not begin until 20 hr after NGF withdrawal. IEG expression generally ends by 24 hr after NGF withdrawal. The IEGs TIS7 and nur77 are not expressed during PCD, yielding a pattern distinct from that following other stimuli. An identical pattern of IEG expression occurs in non-neuronal PC12 cells deprived of serum, although expression begins at 10-14 hr after serum withdrawal. A similar IEG expression pattern was observed in Rat-1 fibroblasts, with various genes expressed 6-18 hr after serum withdrawal. In none of these cell types did expression of the stress related gene Hsp70 change following trophic factor withdrawal. The distinctive pattern of IEG expression described here should facilitate identification of intracellular regulatory signals active during PCD. PMID- 7472490 TI - Neural network of structures in which GABAB receptors regulate absence seizures in the lethargic (lh/lh) mouse model. AB - In previous work we have shown that GABAB receptors are required for expression of absence seizures in the lethargic (lh/lh) mouse model; that lh/lh mice have increased numbers of GABAB binding sites compared to nonepileptic littermates (designated +/+); and that the magnitude of the increased number of GABAB receptors in lh/lh mice correlated positively with the frequency of absence seizures. We performed this study to delineate the neural network in which GABAB receptors regulate absence seizures in lh/lh mice. We designed three successive screens which had to be passed by a candidate neuronal population before it could be considered a member of the neural network in which GABAB receptors regulate absence seizures. First, the neuronal populations in lh/lh mice had to have enriched GABAB binding sites compared to homologous populations in matched nonepileptic controls; baclofen-displaceable 3H-GABA binding was measured in autoradiograms for this screen. Second, the candidate populations had to generate spike-wave discharges (SWDs) during absence seizures in lh/lh mice; bipolar recording electrodes implanted into candidate neuronal structures were used in this screen. Third, the candidate populations had to demonstrate GABAB receptor mediated regulation of absence seizures in lh/lh mice; microinjections of a GABAB agonist [(-)-baclofen] and antagonist (CGP 35348) were used for this screen. In this study we found that anterior ventral lateral thalamic nucleus (VLa), nucleus reticularis thalami (NRT), nucleus reuniens (RE) passed all three screens, and hence are members of the neural network in which GABAB receptors regulate absence seizures in lh/lh mice. PMID- 7472491 TI - Injury-induced upregulation of bFGF and CNTF mRNAS in the rat retina. AB - Focal mechanical injury to the retina has been shown to slow or prevent photoreceptor degeneration near the lesion site in two animal models of retinal degeneration, inherited retinal dystrophy in the Royal College of Surgeons (RCS) and light damage in albino rats. Thus, when injured, the rat retina activates a self-protective mechanism to minimize damage. To identify injury responsive factors and cells, we examined the mRNAs for the following factors and some of their receptors: basic and acidic fibroblast growth factors (bFGF, aFGF) and FGF receptor-1 (FGFR1); ciliary neurotrophic factor (CNTF) and CNTF receptor alpha (CNTFR alpha); brain-derived neurotrophic factor (BDNF) and its receptor trkB; and insulin-like growth factor-1 (IGF-1) and IGFR-1 receptor (IGF-1R). After a single mechanical lesion to the subretinal space and retina, there was a substantial increase in bFGF and CNTF expression that persisted for the entire 10 d period of study. The increase in bFGF mRNA after injury was prompt and great in amplitude, while the elevation of CNTF mRNA was relatively slower. In addition, there was a transient increase in FGFR1 mRNA. In situ hybridizations showed that the elevation of bFGF and CNTF was localized to the vicinity of the lesion. The expression of GFAP (glial fibrillary acidic protein) mRNA also increased in response to injury. These findings strongly suggest that increases in endogenous bFGF and/or CNTF play key roles in injury-induced photoreceptor rescue. PMID- 7472493 TI - Acetylcholine receptor aggregation at nerve-muscle contacts in mammalian cultures: induction by ventral spinal cord neurons is specific to axons. AB - We used a novel mammalian coculture system to study ACh receptor (AChR) redistribution and synaptic structure at nerve-muscle contacts. Ventral spinal cord (VSC) neurons were plated on cultures containing extensive myotubes but few fibroblasts. Neurite-induced redistribution of AChRs occurred within 6 hr after plating neurons and was maximal between 36-48 hr. This AChR redistribution appeared in two patterns: (1) AChR density at sites directly apposed to the neurite where neurites crossed preexisting AChR patches was sharply reduced, (2) Newly aggregated AChRs formed swaths lateral to the neurite path. VSC neurons induced more AChR aggregation than hippocampal, superior cervical ganglion and dorsal root ganglion neurons. The 43 and 58 kDa postsynaptic proteins were colocalized with AChR-enriched domains in all VSC neurite-induced aggregates whereas the colocalization of laminin was variable. Electron microscopy of regions with neurite-induced AChR aggregation showed postsynaptic membrane specializations characteristic of developing synapses and, in older cultures, features of more mature synaptic structure. Thus, the coculture system is useful for studying early stages of neuromuscular junction (NMJ) formation. Neurites in these cocultures were identified as axons or dendrites by morphological criteria and by their immunoreactivity for synaptophysin and phosphorylated heavy neurofilament subunits or for microtubule associated protein 2 (MAP2), respectively. Axons showed a 10-fold higher induction of AChR aggregation than did dendrites. Thus, at least one essential signaling molecule necessary for the induction of AChR aggregation at sites of interaction with muscle appears to be expressed in a polarized fashion in developing VSC neurons. PMID- 7472492 TI - Dye coupling between pyramidal neurons in developing rat prefrontal and frontal cortex is reduced by protein kinase A activation and dopamine. AB - During early postnatal development, lamina II/III pyramidal cells in rat neocortex are extensively coupled via gap junctions. The factors regulating gap junction permeability, as well as the mechanisms underlying the developmental uncoupling process are not understood. To investigate the influence of protein kinase A-mediated phosphorylation on dye coupling in the developing neocortex, pyramidal cells in slices of rat frontal and prefrontal cortex were injected intracellularly with the tracer neurobiotin. Control injections revealed clusters of about 30 dye-coupled neurons. Preincubation with forskolin or direct activation of protein kinase A with Sp-cAMPS reduced the number of coupled cells by about 70%. A significant reduction in dye coupling was also observed following incubation with dopamine. Application of receptor selective agonists and antagonists revealed that the uncoupling was mediated by both dopamine D1 and D2 receptors. The protein kinase A inhibitor Rp-cAMPS reduced the effect of dopamine, suggesting that the neurotransmitter regulates gap junction permeability via protein kinase A activation. In the presence of either forskolin, Sp-cAMPS, or dopamine, neurons displayed a significantly higher input resistance compared to control conditions. During the second postnatal week, transient application of forskolin to single neurons reversibly increased input resistance. At later developmental stages when coupling incidence had declined, this action of forskolin was no longer observed. Our data demonstrate a dependence of gap junction permeability on protein kinase A activity and on dopamine receptor activation in developing rat neocortical neurons. These mechanisms may modulate junctional permeability during the period of circuit formation. PMID- 7472494 TI - Interleukin-1-induced long-lasting changes in hypothalamic corticotropin releasing hormone (CRH)--neurons and hyperresponsiveness of the hypothalamus pituitary-adrenal axis. AB - Hypothalamic CRH neurons that control ACTH secretion from the pituitary gland have secretory terminals in the external zone of the median eminence (ZEME). These neurons can coproduce vasopressin (AVP), a neuropeptide that potentiates the ACTH releasing effects of CRH. Recently, we found increased AVP production in adult rats weeks after single exposure to a stressor, which may play a role in event-induced stress disorders. Here, we describe the long-term changes in the HPA axis of adult male rats following a single exposure to a stressor, the cytokine interleukin-1 beta (IL-1 beta). The effects on storage and release of AVP and CRH were established by quantitative immunocytochemistry, the effects on ACTH and corticosterone responses by radioimmunoassay. Single administration of IL-1 beta (5 micrograms/kg i.p.) induces a delayed (at least 4 d) and a long lasting (at least 3 weeks) increase of vasopressin (AVP) stores in CRH terminals of the ZEME without affecting the CRH stores, and a marked increase of the fraction of CRH terminals that costore AVP. Eleven days after IL-1 beta administration, a second IL-1 beta challenge causes a marked depletion of the AVP stores in the ZEME within 2 hr, which is not seen in rats treated with vehicle 11 d earlier. This is accompanied by twofold higher ACTH and corticosterone responses, as compared to those in vehicle pretreated rats. IL-1 beta-pretreated rats also showed increased ACTH and corticosterone responses to electric footshocks. We conclude that transient activation of the HPA axis by a single administration of IL-1 beta induces a delayed and long-lasting hyperproduction, hyperstorage, and hypersecretion of AVP from hypothalamic CRH neurons that results in hyperresponsiveness of the HPA axis to subsequent stimuli. PMID- 7472495 TI - Cholinergic basal forebrain neurons project to cortical microvessels in the rat: electron microscopic study with anterogradely transported Phaseolus vulgaris leucoagglutinin and choline acetyltransferase immunocytochemistry. AB - Physiological evidence indicates that central cholinergic pathways are involved in the regulation of cerebral cortical blood flow. We investigated the possible contribution of basal forebrain cholinergic neurons from the substantia innominata (SI) to the innervation of cortical microvessels. Basalo-cortical perivascular nerve terminals were detected by light and electron microscopic immunocytochemistry of anterogradely transported Phaseolus vulgaris leucoagglutinin (PHA-L) following its injection in the SI, and were compared to cortical perivascular cholinergic (immunoreactive for choline acetyltransferase (ChAT)) terminals. The basal forebrain origin of cholinergic terminals was evaluated after unilateral ibotenic acid lesion of the SI. PHA-L varicose fibers reached and surrounded microvessels in all cortical subdivisions examined. When studied at the ultrastructural level in the fronto-parietal and perirhinal cortices, perivascular PHA-L nerve terminals were located significantly closer to microvessels in the fronto-parietal than in the perirhinal cortex (respective average distance of 0.98 +/- 0.09 and 1.34 +/- 0.07 micron; p < 0.01). PHA-L and ChAT terminals in the fronto-parietal cortex compared very well in their perivascular distribution but not in the perirhinal cortex. In both cortices, perivascular PHA-L terminals were similar in size to, but engaged more frequently in synaptic contacts than their ChAT counterparts. Following SI lesion, the density of cortical ChAT terminals including those reaching microvessels decreased significantly (56 and 63%, respectively, p < 0.005) in the fronto parietal cortex, while the cortical and perivascular denervations were much less pronounced (26%, not significant and 35%, p < 0.05, respectively) in the perirhinal cortex. These results indicate that basal forebrain neurons project preferentially to fronto-parietal cortical microvessels and further show that a significant proportion of these projections are cholinergic. In addition, the difference in distribution and/or synaptic incidence between perivascular PHA-L and ChAT terminals suggested that noncholinergic SI neurons also contribute to these neurovascular associations, and more so in the perirhinal cortex, as further indicated by the lesion studies. Such finding corroborates recent physiological evidence for a functional innervation of the cortical microvascular bed by SI neurons, a role that might be relevant to the overall pathology of Alzheimer's dementia. PMID- 7472496 TI - Regulated secretion of beta-amyloid precursor protein in rat brain. AB - The beta-amyloid precursor protein (APP) is a ubiquitous, highly conserved secretory glycoprotein that is expressed at high levels in mammalian brain by neurons, astrocytes, and activated microglia. Secreted APP (APPs) is generated by the cleavage of APP within the beta-amyloid (A beta) portion of its ectodomain. The formation and secretion of APPs can be increased by activation of particular neurotransmitter receptors and subsequent protein phosphorylation. We found that tissue slices from rat cortex, hippocampus, striatum, and cerebellum secrete APPs in vitro. APPs secretion was enhanced by electrical stimulation, but was not associated with a general increase in the release of total protein, lactate dehydrogenase (LDH) activity, or neuronal cell adhesion molecules. The pharmacological profile of stimulation-induced APPs secretion suggests complex interactions between muscarinic receptor subtypes in the tissue slices: in the unstimulated state, activation of Muscarinic M1 receptors increased APPs release while nonspecific activation of multiple muscarinic receptors had little effect on APPs release; in electrically stimulated slices, nonspecific inhibition of muscarinic receptors blunted the increase in APPs secretion. The nonspecific muscarinic agonist carbachol increased APPs secretion only in the presence of an M2 receptor antagonist, suggesting that activation of M2 receptors suppresses APPs formation. These data indicate that secretory APP processing in brain includes depolarization-enhanced cleavage of the cell-associated holoprotein within its ectodomain, and that the net effect of depolorization involves several subtypes of acetylcholine receptors. PMID- 7472497 TI - Pharmacological characterization of the rhythmic synaptic drive onto lumbosacral motoneurons in the chick embryo spinal cord. AB - The isolated spinal cord of the chick embryo generates episodes of rhythmic bursting in which sartorius (hip flexor) and femorotibialis (knee extensor) motoneurons exhibit characteristic patterns of activity. At the beginning of each cycle both sets of motoneurons discharge synchronously. Following this brief synchronous activation sartorius motoneurons stop firing at the time of peak femorotibialis activity, producing a period of alternation between the two sets of motoneurons. Intracellular recording from motoneurons has suggested that the pause is mediated by a synaptically induced shunt conductance. However, the pharmacological basis for this shunt and the nature of the excitatory drive to motoneurons is unknown. To address these questions we have investigated the pharmacology of the rhythmic, synaptic drive to lumbosacral motoneurons using local and bath application of several excitatory and inhibitory antagonists, and documenting their effects on motor output in E10-E12 chick embryos. Local application of bicuculline or picrotoxin over sartorius motoneurons abolished the pause in firing recorded from the sartorius muscle nerve. As a consequence, the pattern of sartorius and femorotibialis activity was similar and the motoneurons were coactive. The pause in sartorius firing was shortened following local application of the glycine antagonist strychnine the nicotinic, cholinergic antagonists mecamylamine, and dihydro-beta-erythroidine and several excitatory amino acid antagonists. Application of the GABA uptake inhibitor nipecotic acid depressed the slow potentials and discharge recorded from the sartorius muscle nerve. These findings suggest that the pause is determined primarily by synaptic inputs acting at motoneuron GABAA receptors with contributions from glycinergic, cholinergic, and glutamatergic inputs. The actions of locally applied GABA onto spinal neurons are consistent with these findings because the neurotransmitter depolarizes spinal neurons and reduces their input resistance. Local application of bicuculline, but not strychnine, onto segments containing femorotibialis motoneurons altered the amplitude and duration of femorotibialis discharge and changed the profile of the slow potentials recorded from the muscle nerve. This finding implicates GABAergic inputs in the regulation of femorotibialis discharge. The pause in sartorius firing was still present and a pause in firing appeared in each cycle of femorotibialis discharge following bath application of bicuculline or strychnine. The pause in both sets of motoneurons could be abolished by local application of the NMDA receptor antagonist AP-5 onto the motoneurons, but not by local application of bicuculline. This action of AP-5 was in contrast to its activity in normal Tyrode's solution where it shortened the pause slightly.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 7472498 TI - Extracellular dopamine in the medial preoptic area: implications for sexual motivation and hormonal control of copulation. AB - Dopamine (DA) activity in the medial preoptic area (MPOA) contributes to the control of male rat sexual behavior. We tested (1) whether extracellular DA increases during precopulatory exposure to an estrous female and during copulation, (2) whether exposure to another male increases extracellular DA, (3) whether motor activity during copulation accounts for increased DA levels, and (4) whether concurrent or recent testosterone influences DA levels or copulation in castrates. Extracellular DA and its metabolites in male rats' MPOA were measured using microdialysis. DA level increased during precopulatory exposure to the female in all animals that subsequently copulated; this included all intact animals, all testosterone-treated castrates, and 9 of 14 1-week castrates treated with oil vehicle. DA levels did not increase in any animal that subsequently failed to copulate, including the remaining 1-week, and all 2-week, vehicle treated castrates. When the barrier was removed and the animals were allowed to copulate, levels of DA and its metabolites continued to rise in intact males and in castrates that copulated. The DA response to the estrous female could not be attributed to nonsexual social stimuli, since exposure to another male was ineffective. The DA response to copulation could not be attributed primarily to motor activity, since animals running voluntarily in a running wheel did not show significantly increased DA. These and previous data suggest that DA released in the MPOA in response to an estrous female may contribute to sexual motivation and copulatory proficiency. Testosterone may promote copulation in part through permissive actions on dopamine release. PMID- 7472499 TI - Attention and target selection for smooth pursuit eye movements. AB - Two rhesus monkeys were trained to track a small moving target in the presence of a moving distractor. The target and distractor were distinguished by their color. Smooth pursuit eye movements were quantified in terms of the latency of the eye movement and the open-loop eye acceleration profile. Smooth pursuit latencies for single targets were on the order of 100 msec. When the target was paired with a distractor moving in the same direction as the target, pursuit latencies decreased to roughly 85 msec. When the target was paired with a distractor moving in the opposite direction, pursuit latencies increased to roughly 150 msec. The motion of the distractor had no significant effect on the eye acceleration profile. Experiments were performed to dissociate visual search for the target from pursuit initiation by providing a spatial cue rather than the color cue. These experiments showed that visual search necessarily preceded pursuit initiation only when the distractor moved in the opposite direction relative to the target. In this case, visual search contributed about 25 msec to the overall latency of pursuit. Control experiments showed that the monkey need not attend to the distractor in order for it to influence the latency of pursuit. A network model was developed in which units that represent the motions of the target and distractor compete against one another. Attention serves to bias the outcome of this competition toward the direction of the selected target. The performance of this network exhibits a striking parallel to the effect of the distractor on smooth pursuit latency. PMID- 7472500 TI - Protein kinase C involvement in homologous desensitization of delta-opioid receptor coupled to Gi1-phospholipase C activation in Xenopus oocytes. AB - We have developed the coexpression system of both delta-opioid receptor (DOR1) and M2-muscarinic receptor (M2) which mediate agonist-evoked currents due to common post-receptor mechanisms including Gi1 and phospholipase C (PLC) activation in Xenopus oocytes reconstituted with Gi1 alpha. The DOR1-currents by 100 nM D-Ser2-leu-enkephalin-Thr6 (DSLET) were selectively desensitized by 10 nM phorbol 12-myristate 13-acetate (PMA). The PMA-desensitization of DSLET-currents was abolished in the presence of calphostin C, a protein kinase C inhibitor, or reversed by an intracellular injection of calcineurin, a protein phosphatase 2B. When a higher concentration (3 microM) of DSLET was used, DSLET-currents were rapidly desensitized by repeated challenges of DSLET itself. However, repeated challenges of 10 microM ACh caused no influence on such DSLET- or M2-currents. The desensitization of DSLET-currents was selectively reversed by protein kinase C inhibitors. Similar results were also obtained with various delta-opioid agonists. These results suggest that protein kinase C is involved in the homologous desensitization of delta-opioid receptors. PMID- 7472501 TI - Reevaluation of the growth-permissive substrate properties of goldfish optic nerve myelin and myelin proteins. AB - To determine whether optic nerve myelin of goldfish carries mammalian-like neurite growth inhibitory proteins which can be neutralized by the antibody IN-1, myelin fractions of fish optic nerves were used as substrates for fish retinal ganglion cell axons and rat dorsal root ganglia (DRG). Axonal growth was monitored and compared with that of IN-1 treated preparations. Growth of fish retinal axons and rat DRG neurites was substantial on goldfish optic nerve myelin and no improvement was observed with IN-1. In contrast, rat CNS myelin allowed only poor growth, and number of axons and length of DRG neurites increased significantly with IN-1. In addition, proteins of fish optic nerve myelin and bovine CNS myelin were extracted, reconstituted in liposomes and applied to growth cones. When goldfish myelin proteins in liposomes were seeded onto growth cones, 77% of fish and 89% of rat DRG growth cones continued to elongate, and the proportion of elongating fish growth cones (80%) did not significantly change when liposomes were pretreated with IN-1. But 73% of fish and 93% of rat growth cones collapsed with liposomes containing proteins from bovine CNS myelin. Upon IN-1 treatment, only 24% of fish growth cones collapsed. Thus, axon growth in vitro indicates that goldfish optic nerves, which permit successful axon regeneration in vivo, lack mammalian-like neurite growth inhibitors which are neutralized by IN-1. PMID- 7472502 TI - The immunosuppressant FK506 increases the rate of axonal regeneration in rat sciatic nerve. AB - The axonal regenerative properties of the new immunosuppressant drug FK506 (tacrolimus) are further explored in this continuing study. In an initial report (Gold et al., 1994a), we described the ability of FK506 to reduce the time until return of function in the hind feet of rats following a sciatic nerve crush. In the present study, we examined the morphological correlate underlying this enhancement of functional recovery. In rats receiving daily subcutaneous injections of FK506 (1.0 mg/kg) for 18 d following a sciatic nerve crush the regenerating axons appeared larger in size compared to saline-injected control animals. Morphometric analysis of axonal calibers in the soleus nerve demonstrated that mean axonal areas for the largest 30% of axons were increased over axotomized control values by 93% in the FK506-treated animals. Next, the rate of axonal regeneration was determined by radiolabeling the L5 dorsal root ganglion (DRG) at 9 and 14 d following axotomy. Regression analysis of the outgrowth distances for sensory axons between 10 and 15 d revealed a 16% increase in regeneration rate. Electron microscopy of intramuscular nerve branches in the interosseus muscles confirmed that the axons in the FK506-treated animals were further advanced toward their targets; in some instances, axons were shown to reinnervate muscle spindles. The results are discussed in terms of the known ability of FK506 to inhibit the activity protein phosphatase 2B (calcineurin). PMID- 7472503 TI - Transient versus persistent functional and structural changes associated with facilitation of Aplysia sensorimotor synapses are second messenger dependent. AB - Increases in activity of both protein kinase A (PKA) and protein kinase C (PKC) contribute to short-term facilitation of Aplysia sensorimotor synapses evoked by serotonin (5-HT). We report here that increasing levels of cAMP in sensory neurons evokes increases in both synaptic efficacy and in the number of sensory neuron varicosities contacting the major axons of motor cell L7 at intermediate times (3 hr) that persist for 24 hr. Treatment with phorbol esters results in a large transient increase in synaptic efficacy that is accompanied by a large transient increase in the number of sensory neuron varicosities with the newest varicosities most susceptible to elimination. The reversal of the synaptic facilitation and the structural changes does not appear to be the result of long term inhibitory actions of persistent PKC activation by phorbol esters, since changes in synaptic efficacy can be evoked by additional applications of either phorbol esters or 5-HT. The short-lived changes in structure evoked by phorbol esters occur in preexisting sensory neurites and not by new growth, since increases in PKC activity with phorbol esters lead to reductions in neurite extension and to retractions by sensory neuron growth cones. The action of phorbol esters on growth cone extension is reversible with washout. The results suggest that increases in PKA and PKC activities by 5-HT contribute to short (minutes) and intermediate (hours) forms of facilitation of sensorimotor synapses while increases in PKA activity also mediate long-term (days) maintenance of synaptic facilitation. PMID- 7472504 TI - Long-term potentiation of glutamatergic pathways in the lamprey brainstem. AB - Plasticity of synaptic transmission has been investigated in the lamprey brainstem. In this preparation, neurons of the vestibular nuclei make monosynaptic connections to reticulospinal neurons of the posterior (PRRN) and middle (MRRN) rhombencephalic reticular nuclei. This glutamatergic projection shows a long-lasting enhancement in efficacy following a brief high frequency (50 Hz) train of stimuli (tetanus). Enhancement was found on the inputs from the octavomotorius intermediate (nOMI) and octavomotorius posterior (nOMP) nuclei to the reticulospinal neurons of the PRRN and MRRN, and persisted for the entire duration of the recordings. The enhancement was limited to the pathway that received tetanic stimulation for the inputs to any given reticulospinal neuron. It depended upon the activation of postsynaptic processes at least in part; dialysis of the reticulospinal neuron by recording with a whole-cell patch pipette prevented its induction. This dialysis-dependent abolition of enhancement was shown to be dependent on a change in Ca2+ concentration in the postsynaptic neuron. The enhancement was not affected by blockade of NMDA receptors with D,L-2 amino-5-phosphonopentanoate (AP5) but was prevented by the metabotropic glutamate receptor (mGluR) antagonist (R,S)-alpha-methyl-4-carboxyphenylglycine (MCPG). In conclusion, this study demonstrates that vestibular inputs to reticulospinal neurons are capable of undergoing long-term potentiation (LTP) and that this LTP shows synapse specificity. Furthermore, this LTP is activated by synaptic stimulation of a subtype of the m-GluR family and its induction is mediated by changing Ca2+ concentrations in the postsynaptic neuron. PMID- 7472505 TI - Regulation of BDNF and trkB mRNA in rat brain by chronic electroconvulsive seizure and antidepressant drug treatments. AB - The influence of chronic electroconvulsive seizure (ECS) or antidepressant drug treatments on expression of brain-derived neurotrophic factor (BDNF) and its receptor, trkB, was examined by in situ hybridization and Northern blot. In frontal cortex, acute ECS increased BDNF mRNA approximately twofold, an effect significantly augmented by a prior course of chronic ECS treatment (10 d). In the hippocampus, the influence of chronic ECS varied between the major subfields. In the dentate gyrus granule cell layer, chronic ECS decreased the acute induction of BDNF and trkB mRNA by approximately 50%, but prolonged their expression: levels remained elevated two- to threefold 18 hr later after the last chronic ECS treatment, but returned to control 18 hr after acute ECS. In CA3 and CA1 pyramidal cell layers, chronic ECS significantly elevated the acute induction of BDNF, and tended to prolong the expression of BDNF and trkB mRNA. A similar effect was observed in layer 2 of the piriform cortex, where chronic ECS significantly increased the acute induction and prolonged the expression of BDNF and trkB mRNA. Chronic (21 d), but not acute (1 d), administration of several different antidepressant drugs, including tranylcypromine, sertraline, desipramine, or mianserin, significantly increased BDNF mRNA and all but mianserin increased trkB mRNA in hippocampus. In contrast, chronic administration of nonantidepressant psychotropic drugs, including morphine, cocaine, or haloperidol, did not increase levels of BDNF mRNA. Furthermore, chronic administration of ECS or antidepressant drugs completely blocked the down regulation of BDNF mRNA in the hippocampus in response to restraint stress. The enhanced induction and prolonged expression of BDNF in response to chronic ECS and antidepressant drug treatments could promote neuronal survival, and protect neurons from the damaging effects of stress. PMID- 7472506 TI - Synaptic plasticity in the basolateral amygdala induced by hippocampal formation stimulation in vivo. AB - Several studies suggest that axonal projections from the hippocampal formation (HF) to the basolateral amygdala (BLA) play a role in Pavlovian fear conditioning to contextual conditional stimuli. We have used electrophysiological techniques to characterize neuronal transmission in these projections in urethane anesthetized rats. Single-pulse electrical stimulation of the ventral angular bundle (VAB), which carries projections from the HF to the BLA, reliably evoked a biphasic extracellular field potential in the BLA that consisted of an early, negative and a late, positive component. The negative component of the field potential occurred at a short latency (3-8 msec), was both temporally and spatially correlated with VAB-evoked multiple-unit discharges in the BLA, and exhibited properties typical of a monosynaptic response. Infusion of lidocaine or glutamate receptor antagonists into the BLA attenuated VAB-evoked field potentials, indicating that they are generated by local synaptic glutamatergic transmission. Both paired-pulse stimulation and brief trains of high-frequency stimulation (HFS) induced a short-lasting facilitation of BLA field potentials, whereas longer and more numerous trains of HFS produced an enduring, NMDA receptor-dependent long-term potentiation (LTP) of the potentials. The induction of LTP was accompanied by a decrease in paired-pulse facilitation (PPF), suggesting a presynpatic modification underlying its expression. Electrolytic lesions placed in regions of the HF that project to the BLA or excitotoxic lesions placed in the BLA eliminated Pavlovian fear conditioning to a contextual conditional stimulus. The critical role of both structures in context conditioning implicates plasticity at HF-BLA synapses in this form of learning. PMID- 7472507 TI - Sympathetic axons pathfind successfully in the absence of target. AB - To determine whether sympathetic axons require the presence of a peripheral target to grow to the correct destination, we examined the developing footpad innervation in tabby mutant mice which lack sweat glands. Despite the absence of sweat glands, noradrenergic sympathetic axons are transiently present in the presumptive target area and avoid the more distal epidermal/dermal domain occupied by sensory axons. Since sympathetic axon pathfinding was not dependent upon the target tissue, we compared the subsequent development of sweat gland axons in tabby footpads with that in control footpads. In wild-type mice, the gland-associated axonal plexus expands considerably as the secretory tubule enlarges and coils. This expansion, however, does not occur in tabby mice. The sweat gland innervation of wild-type mice loses catecholamines and acquires AChE activity and vasoactive intestinal peptide immunoreactivity. In tabby mutant mice, catecholaminergic fibers remain in the glandless footpads for 2 weeks and fail to acquire AChE or vasoactive intestinal peptide. In contrast to the altered development of gland innervation in tabby, the development of the innervation of footpad blood vessels was unaffected. Our observations indicate that the target is not required to direct sympathetic axons to the presumptive gland region of the footpad. In the absence of the target tissue, however, gland-targeted sympathetic axons retain an immature morphology and transmitter phenotype and then disappear. PMID- 7472508 TI - Persistent cardiovascular and behavioral nociceptive responses to subcutaneous formalin require peripheral nerve input. AB - Hindpaw injection of formalin produces acute (Phase 1) and persistent (Phase 2) nociceptive behaviors. This model has provided critical evidence supporting a contribution of central sensitization (hyperexcitability of spinal neurons) to the expression of persistent pain. Here, we evaluated the contribution of ongoing peripheral nerve inputs to Phase 2 pain responses. In addition to pain behavior (flinching), we measured formalin-evoked increases in arterial pressure and heart rate; these cardiovascular responses were also biphasic in nature. The arterial pressure response correlated highly with behavior, and was dependent on formalin concentration (0.625-5.0%), indicating that it was largely driven by noxious input. Lightly anesthetized (0.7% halothane) rats exhibited robust increases in blood pressure in the absence of pain behavior, indicating cardiovascular responses did not reflect somatomotor-cardiovascular coupling. Animals obtained from Charles River exhibited slightly larger Phase 2 flinching and heart rate responses compared to those obtained from Bantin and Kingman, suggesting cardiovascular-related pain responses can vary with the source of animal. We next evaluated the contribution of ongoing peripheral nerve activity to the expression of the Phase 2 pressor, tachycardia, and flinch responses. After Phase 1 subsided, but before Phase 2 began, we locally anesthetized the ipsilateral or contralateral (control) hindpaw with a hydrophilic lidocaine derivative, QX-314 (2%). Intraplantar QX-314 blocked Phase 2 pressor, tachycardia and behavioral responses only when injected into the paw that received formalin (2.5% or 10.0%). We conclude that persistent ongoing activity in peripheral afferent fibers during Phase 2 is required for the persistent pain evoked by formalin. PMID- 7472509 TI - Cloning and characterization of two related serotonergic receptors from the brain and the reproductive system of Aplysia that activate phospholipase C. AB - Serotonin (5-HT) plays important roles in various behavioral and physiological processes in Aplysia californica. These include feeding, locomotion, circadian rhythm, learning and memory, synaptic plasticity, and synaptic growth. Serotonin modulates these various functions by interacting with different 5-HT receptor subtypes that are coupled to various second-messenger systems. We report here the isolation and characterization of the first two serotonergic receptors from Aplysia californica, Ap5-HTB1 and Ap5-HTB2, using a strategy based on the amino acid sequence homology among G-protein-coupled biogenic amine receptors. Ap5-HTB1 and Ap5-HTB2 are both intronless and highly homologous to each other, sharing 79.5% sequence identity at the amino acid level. Sequence comparison reveals that these receptors are 33.1 to 23.3% identical to the following 5-HT receptors: 5 HTdro1 > 5-HT6 > 5-HTlym > mouse 5-HT1B > 5-HTdro2A > mouse 5-HT7 > rat 5-HT2A. Both Ap5-HTB1 and Ap5-HTB2 encode functional 5-HT receptors. When expressed in cultured cells, these receptors stimulate phospholipase C in response to 5-HT in a dose-dependent manner. This stimulation can be blocked by specific 5-HT receptor antagonists. Using RT-PCR and Western blot analysis, we have detected these receptors in the CNS (Ap5-HTB2) and in the reproductive system (Ap5-HTB1). The nucleotide sequences of Ap5-HTB1 and Ap5-HTB2 were submitted to GenBank; the accession numbers are L43557 and L43558, respectively. PMID- 7472510 TI - Persistent activation of cAMP-dependent protein kinase by regulated proteolysis suggests a neuron-specific function of the ubiquitin system in Aplysia. AB - In response to the facilitating neurotransmitter serotonin (5-HT), the cAMP dependent protein kinase (PKA) acquires a special mnemonic characteristic in Aplysia sensory neurons. PKA becomes persistently activated at basal cAMP concentrations owing to a decreased regulatory (R) to catalytic (C) subunit ratio. We previously implicated ubiquitin-mediated proteolysis in this selective loss of R. Here we show that ubiquitin (Ub), Ub-conjugates and proteasomes are present in cell bodies, axon, neuropil and nerve terminals of Aplysia neurons. Because R subunits are not decreased in muscle exposed to 5-HT, comparison of the two tissues provides a tractable approach to determine how the Ub pathway is regulated. We compared the structure of M1, the muscle-specific R isoform, to that of N4, a major neuronal R isoform, to rule out the possibility that the differences in their stability result from differences in structure. We present evidence that N4 and M1 are encoded by identical transcripts; they also behave similarly as protein substrates for the Ub pathway in extracts of the two tissues. Nervous tissue contains 20-times more free Ub, but we present evidence that the susceptibility of R subunits to degradation in neurons relative to muscle results from the greater capacity of neurons to degrade ubiquitinated proteins through the proteasome. Thus, factors that regulate the activity of proteasomes could underlie the enhanced degradation of R subunits in long-term sensitization. PMID- 7472511 TI - The behavior of APO-SUS rats in animal models with construct validity for schizophrenia. AB - Schizophrenic patients are known to suffer from a number of information processing disturbances, including deficits in both prepulse inhibition of startle and latent inhibition. Since these behavioral phenomena can also be observed in animals, they represent an ideal starting point for developing animal models having construct validity for specific deficits observed in schizophrenia. The principal question is how to induce a condition in animals most similar to the schizophrenic deficit. In the present study, we have selected rats on the basis of their response to an open filed or to the dopaminergic agonist apomorphine, and evaluated their prepulse inhibition and latent inhibition. We used three different selection procedures (open field selection for novelty response, gnawing cage selection for apomorphine response, and pharmacogenetic selection for apomorphine response). The results show that, irrespective of the selection procedure used, rats with a high response to novelty or apomorphine susceptible (collectively called APO-SUS rats) show diminished prepulse inhibition of the acoustic startle response as compared to rats with a low response to novelty or apomorphine unsusceptible (collectively called APO-UNSUS rats). This difference was apparent only at low prepulse intensities. Moreover, these APO-SUS rats show diminished latent inhibition in a conditioned taste aversion paradigm as compared to APO-UNSUS rats. Given the fact that the pharmacogenetically bred APO-SUS rats show several central nervous, endocrinological, and immunological similarities to schizophrenic patients, they are hypothesised to represent an interesting nonpharmacological animal model for schizophrenia-prone patients. PMID- 7472512 TI - Biphasic changes in locomotor behavior and in expression of mRNA for NGFI-A and NGFI-B in rat striatum following acute caffeine administration. AB - The time course of expression of mRNA for NGFI-A and NGFI-B after a single intraperitoneal injection of saline or caffeine was examined using in situ hybridization. Administration of a high dose of caffeine (100 mg/kg) decreased locomotor behavior and increased NGFI-A and NGFI-B mRNA in the entire striatum. A lower dose of caffeine (50 mg/kg) caused a weak enhancement of both messages, which was confined to the lateral part of caudate-putamen. This dose increased horizontal, but not vertical, movement. In rats treated with the lowest dose of caffeine (25 mg/kg), the expression of both investigated genes tended to be lower than for saline-treated rats and both horizontal and vertical locomotor activity increased markedly. The reduction in the number of labeled neurons seemed to occur predominantly in enkephalin-containing neurons, which coexpress adenosine A2A receptors and dopamine D2 receptors. The decrease of mRNA for NGFI-A, NGFI-B, and jun B caused by caffeine (25 mg/kg) could be mimicked by the D2 agonist quinpirole (1 and 3 mg/kg). Moreover, caffeine could significantly decrease the expression seen following treatment with the D2 antagonist raclopride (2 mg/kg). In addition, in the parietal cortex, 25 mg/kg of caffeine caused a significant elevation of both examined immediate early genes. Thus, biphasic changes in locomotion induced by caffeine are paralleled by biphasic changes in mRNA for NGFI-A, NGFI-B, and jun B. The results also provide additional support for a functionally important interaction between adenosine and dopamine D2 receptors. PMID- 7472513 TI - Islet amyloid polypeptide (amylin) is expressed in sensory neurons. AB - Islet amyloid polypeptide (IAPP) or amylin is a hormone candidate predominantly expressed in insulin cells. A role for IAPP in the regulation of glucose homeostasis and the development of non-insulin-dependent diabetes mellitus has been proposed. IAPP is structurally related to the sensory neuropeptide calcitonin gene-related peptide. In the present study, using in situ hybridization, immunocytochemistry, and immunochemistry, the expression of IAPP in sensory neurons in the rat was investigated. IAPP was expressed in a population of small- to medium-sized nerve cell bodies in dorsal root ganglia from all levels and in the jugular-nodose and trigeminal ganglion; IAPP expressing nerve cell bodies constituted a subpopulation of those expressing calcitonin gene-related peptide. In addition, IAPP-like immunoreactivity occurred in nerve cell bodies storing substance P and pituitary adenylate cyclase activating polypeptide. IAPP-immunoreactive nerve fibers were encountered in the dorsal horns of the spinal cord, and to a lesser extent in peripheral tissues receiving sensory innervation; IAPP-immunoreactive fibers constituted a subpopulation of those containing calcitonin gene-related peptide and/or substance P. The immunochemical determinations demonstrated a low level of IAPP like immunoreactivity in the dorsal root ganglia and spinal cord, which chromatographically coeluted with authentic rat IAPP. We conclude that IAPP is expressed in sensory neurons, thus being a novel sensory neuropeptide candidate for which a physiological role remains to be identified. PMID- 7472514 TI - Substance P induced by peripheral nerve injury in primary afferent sensory neurons and its effect on dorsal column nucleus neurons. AB - Using in situ hybridization and the retrograde tracer, Fluorogold, we examined the expression of preprotachykinin (PPT) mRNA in the rat dorsal root ganglion neurons projecting to the gracile nucleus. Seven days after unilateral sciatic nerve transection, some medium- to large-sized neurons in the rat dorsal root ganglia projecting to the gracile nucleus express PPT mRNA, whereas very few gracile nucleus-projecting neurons on the contralateral side express PPT mRNA. Immunohistochemistry revealed an increase in substance P (SP) immunoreactivity in the gracile nucleus and large myelinated fibers in the dorsal root 2 weeks after unilateral sciatic nerve transection. The results suggest that medium to large DRG cells that project to the gracile nucleus express PPT mRNA de novo in response to peripheral nerve injury, and increased SP is transported to the gracile nucleus through large myelinated fibers. To determine whether the increased SP might affect the excitability of the gracile nucleus neurons postsynaptically, Fos expression after electrical stimulation of the injured sciatic nerve was examined. Multiple injections of the NK-1 receptor antagonist, CP-96,345, suppressed stimulus-induced Fos expression in gracile nucleus neurons including thalamic relay neurons. The inactive enantiomer, CP-96,344, had no effect on stimulus-induced Fos expression. These data indicate that the de novo synthesized SP in the lesioned primary afferent neurons may be involved in an augmentation of excitability in the dorsal column-medial lemniscus sensory pathway. This hyperexcitability may play a role in the pathogenesis of abnormal neuropathic sensations following peripheral nerve injury. PMID- 7472515 TI - Effects of delayed visual information on the rate and amount of prism adaptation in the human. AB - Accurate reaching towards a visual target is initially disturbed when the visual field is displaced by prisms, but recovers with successive trials. To determine how the improvement depends on the visual error signals associated with the motor output, the time course of prism adaptation was studied with delayed visual information on the error. Subjects were trained to reach rapidly at a target on a tangent screen. Vision was always blocked during the movement, and allowed again only after the index finger touched the screen. One experiment consisted of three sets of 30 trials. In the first set, the subject wore no prisms and vision was allowed without delay. In the second, the visual field was displaced by prisms, and vision was available only after a delay period of 0-10,000 msec while the subjects maintained their final pointing position. Initially, the subject misreached the target by about the amount of visual displacement (60 mm). Errors decreased with trials by an amount proportional to the error in the preceding trial. The rate of decrease of error was generally largest when the delay was 0 msec, became significantly smaller when the delay was 50 msec, and showed only gradual change with longer delays. In the third set, the subject wore no prisms and vision was allowed without delay. Initial misreaching in the direction opposite to the visual displacement, reflecting the amount of adaptation in the second set, was generally largest with no delay (median of 46 mm) and significantly smaller with 50 msec and longer delays (17-33 mm).(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7472516 TI - Behavioral role for nitric oxide in chemosensory activation of feeding in a mollusc. AB - A role for the NO-cGMP pathway in mediating chemosensory activation of feeding is suggested by intense NADPH diaphorase staining observed in nerve fibers that project from sensory cells in the lips to the CNS and by the presence in the CNS of a NO-activated guanylyl cyclase. In preparations reduced to isolated lips and CNS, intracellular recordings were made from motoneurons driven by the interneurons of the central pattern generator (CPG) for feeding. Fictive feeding in such preparations can be recorded from these motoneurons following the application of sucrose to the lips. Sucrose activation of fictive feeding is inhibited by the NO scavenger hemoglobin, the NO synthase inhibitor N omega-Nitro L-Arginine Methyl Ester (L-NAME) and by methylene blue, an inhibitor of guanylyl cyclase. Fictive feeding in isolated lip-CNS preparations can be activated without sucrose by superfusion of NO donor molecules such as SNAP and hydroxylamine and by the nonhydrolyzable analog of cGMP, 8-bromo-cGMP. The feeding CPG can also be activated centrally by depolarizing a modulatory interneuron, the slow oscillator (SO). When the CPG is activated in this way, fictive feeding is not susceptible to inhibition by hemoglobin, the most potent of the inhibitors of sucrose-activated fictive feeding. Behavioral experiments on intact snails confirm the findings from in vitro experiments and show that hemoglobin prevents feeding and methylene blue significantly delays the onset of feeding. These results indicate (1) that NO is a putative chemosensory transmitter in the snail L. stagnalis, (2) that the NO-cGMP pathway can mediate chemosensory activation of specific patterns of centrally generated behavior, (3) that NO is not involved in transmission within the central network of neurons responsible for the behavior, and more generally (4) that a freely diffusing and highly reactive gaseous signalling molecule can have restricted and specific behavioral functions. PMID- 7472517 TI - The chronology of oligodendrocyte differentiation in the rat optic nerve: evidence for a signaling step initiating myelination in the CNS. AB - In order to determine the signals that initiate axon myelination in the CNS, we have chronicled the differentiation of oligodendrocytes in the rat optic nerve and related this to the time course and spatial gradient seen for optic axon myelination. By using markers specific to the varying stages of oligodendrocyte differentiation we found that oligodendrocyte progenitor cells, present throughout the length of the nerve at postnatal day 2, mature into GC+ oligodendrocytes in a chiasm to eye progression. This gradient along the nerve of oligodendrocyte differentiation continues with oligodendrocytes near the chiasm expressing the genes and encoded proteins to MBP and PLP 3 d before oligodendrocytes near the eye. Although oligodendrocyte differentiation and maturation occurs in a chiasm to eye gradient along the nerve, optic axon segments near the eye are ensheathed with myelin before segments near the chiasm. This suggests that the myelination of optic axons is initiated by a signaling step that is independent of oligodendrocyte differentiation and is stronger near the eye than the chiasm region of the nerve. By examining proposed axonal signals, we found that the onset of myelination is independent of the electrical activity of an axon but can be correlated to the size of an axon. PMID- 7472518 TI - How retinal microcircuits scale for ganglion cells of different size. AB - Ganglion cell receptive field centers are small in central retina and larger toward periphery. Accompanying this expansion, the distribution of sensitivity across the centers remain Gaussian, but peak sensitivities decline. To identify circuitry that might explain this physiology, we measured the density of bipolar cell synapses on the dendritic membrane of beta (X) and alpha (Y) ganglion cells and the distribution of dendritic membrane across their dendritic fields. Both central and peripheral beta cells receive bipolar cell synapses at a density of approximately 28/100 microns2 of dendritic membrane; central and peripheral alpha cells receive approximately 13/100 microns2. The distribution of dendritic membrane across the dendritic field is dome-like; therefore, the distribution of bipolar cell synapses is also dome-like. As the dendritic field enlarges, total postsynaptic membrane increases with field radius, but only linearly. Consequently, density of postsynaptic membrane in the dendritic field declines, and so does density of synapses within the field. The results suggest a simple model in which the receptive field center's Gaussian profile and peak sensitivity are both set by the density of bipolar cell synapses across the dendritic field. PMID- 7472519 TI - A single histidine residue is essential for zinc inhibition of GABA rho 1 receptors. AB - The GABA rho 1 subunit, cloned from a human retina library, can form homooligomeric receptors with properties similar to GABAc receptors characterized in retinal cells. The divalent cation Zn2+, abundant in the CNS and retina, was found to inhibit GABA rho 1 receptors in a voltage-independent manner. Varying the extracellular pH from 7.4 to 5.6 significantly reduced this inhibitory effect. This pH profile suggested that one or more histidine residues might play a role in the interaction between Zn2+ and the GABA rho 1 receptor. Site-directed mutagenesis revealed that a single histidine residue (His 156) in the putative extracellular domain of rho 1 was critical for Zn2+ sensitivity. Substitution of this amino acid with tyrosine (H156Y) created a functional GABA receptor with agonist and channel properties indistinguishable from wildtype. However, the H156Y mutant was insensitive to Zn2+, even at concentrations as high as 1 mM. Mutation to aspartic acid, an amino acid that can interact with Zn2+ in other proteins, preserved sensitivity to Zn2+ but abolished the pH-dependent effect. This histidine residue is also involved in Ni2+ and Cd2+ interaction since the H156Y mutation completely suppressed the inhibition effects of these two cations. These data demonstrate that an extracellular histidine residue is critical for transition metal cation sensitivity of GABA rho 1 receptors. PMID- 7472520 TI - NMDA-mediated modulation of gamma-aminobutyric acid type A receptor function in cerebellar granule neurons. AB - GABAA receptors are ligand-gated CI- ion channels with multiple clinically relevant drug-recognition sites. We have previously shown that stimulation of N methyl-D-aspartic acid (NMDA)-specific glutamate receptors quantitatively alters selected GABAA receptor subunit mRNAs and proteins in primary cultures of rat cerebellar granule neurons. We used whole-cell recordings of GABA-elicited CI- currents and flunitrazepam binding experiments in granule cell cultures maintained in low K+ (12.5 mM), cells maintained in low K+ and treated with a single dose of NMDA (10 microM), and cell cultures maintained in depolarizing concentrations of K+ (25 mM). The EC50 obtained from the dose-response curves for GABA in eliciting a maximal response was comparable in neurons maintained in high K+ or in low K+ and treated with a single dose of NMDA, but that it increased significantly in cells maintained in low K+. The potentiation of GABA-gated CI- currents by flunitrazepam increased significantly, while the negative allosteric modulator methyl-6,7-dimethoxy-4-ethyl-beta-carboline-3-carboxylate (DMCM) was significantly more effective in cultures either maintained in high K+ or treated with NMDA. This was coincident with a twofold increase in the Bmax associated with flunitrazepam binding. To further characterize the receptor assemblies present in the depolarization and NMDA induced paradigms, the Zn(2+)-induced inhibition of GABA-gated CI- currents was reduced as was the inhibition mediated by furosemide. Our data indicate that GABAA receptor assemblies alter their composition in response to excitatory afferent receptor stimulation. PMID- 7472521 TI - Glutamate and non-glutamate receptor mediated toxicity caused by oxygen and glucose deprivation in organotypic hippocampal cultures. AB - In vitro ischemia models have utilized oxygen, or oxygen and glucose deprivation to simulate ischemic neuronal injury. Combined oxygen and glucose deprivation can induce neuronal damage which is in part mediated through NMDA receptors. Severe oxygen deprivation alone however can cause neuronal injury which is not NMDA mediated. We tested the hypothesis that NMDA, or non-NMDA receptor mediated mechanisms may predominate, to induce neuronal injury following severe oxygen deprivation depending on the presence of glucose. We found that NMDA receptor blockade using dizocilpine (MK-801), DL-2-amino-5-phosphonovaleric acid (APV), or CGS 19755, was highly effective in reducing CA1 injury in organotypic hippocampal cultures, caused by complete oxygen and glucose deprivation. Complete oxygen deprivation alone however, caused CA1 neuronal injury which was not diminished using NMDA receptor blockade alone with MK-801 or APV, or in combination with AMPA/kainate receptor blockade using 6-cyano-7-dinitroquinoxalone-2,3-dione (CNQX). Neuronal protective strategies which act primarily through non-glutamate dependent mechanisms, including hypothermia, low chloride and calcium, and the free radical scavenger, alpha-phenyl-tert-butyl nitrone (PBN), provided neuronal protection against complete oxygen, as well as combined oxygen/glucose deprivation. Raising the pH using Hepes buffer during complete oxygen deprivation did not result in neuronal protection by NMDA receptor blockade. Partial oxygen deprivation alone, partial oxygen deprivation combined with glucose deprivation, glucose deprivation alone, and also glutamate exposure, all produced neuronal damage that was reduced by NMDA receptor blockade. The presence of glucose during complete oxygen deprivation appears to prevent glutamate receptor blockade from reducing neuronal injury in organotypic hippocampal cultures. PMID- 7472522 TI - Cell surface morphology identifies microglia as a distinct class of mononuclear phagocyte. AB - To investigate differences among brain-derived microglia and other classes of immune cells, we compared the morphologies and growth properties of mononuclear phagocytes isolated from tissues of the newborn rat. Scanning EM shows that microglia from postnatal rat brain are covered with spines (typically > 20 per cell body) in a distinctive manner which contrasts the smooth surfaces of bone marrow cells and the ruffled surfaces of tissue macrophages from spleen, liver, and peritoneum. The spine-bearing surface of microglia is a specific cell marker, for it does not change with age or after exposure to cytokines or other immunostimulants. Approximately 99% of mononuclear phagocytes cultured from normal adult rat brain are spinous microglia. Five days after injury to rat brain, cells at sites of Wallerian degeneration are essentially all spinous ones while nearly 30% of cells found within areas of infarction or penetrating trauma are invading macrophages. In a similar way, nearly all cells isolated from normal, postmortem adult human brain are spine-bearing microglia (> 99% homogeneity). Brains from patients with amyotrophic lateral sclerosis contain only spinous microglia whereas cells from HIV-1 infected brains include significant numbers of invading ruffled macrophages. Cultured microglia, unlike cultured bone marrow precursors, monocytes, or tissue macrophages, spontaneously develop long, thin processes that extend hundreds of microns in length. Microglia retract these processes after exposure to fetal bovine serum, laminin, or such immunostimulants as recombinant murine interferon gamma (rmIFN gamma) and lipopolysaccharide. Of all types of mononuclear phagocytes tested, only microglia differentiate into quiescent ramified cells when in contact with astrocytes. Thus, microglia represent a unique class of cell maintained, in part, by astroglia as dormant, ramified mononuclear phagocytes in mature CNS. Application of cell surface criteria described here will allow study of distinct populations of mononuclear phagocytes associated with neurologic disorders. PMID- 7472523 TI - Bcl-2 overexpression prevents motoneuron cell body loss but not axonal degeneration in a mouse model of a neurodegenerative disease. AB - Bcl-2 and its analogs protect different classes of neurons from apoptosis in several experimental situations. These proteins may therefore provide a means for treatment of neurodegenerative diseases. We examined the effects of Bcl-2 overexpression in a genetic mouse model with motor neuron disease (progressive motor neuronopathy/pmn). Pmn/pmn mice lose motoneurons and myelinated axons, and die at 6 weeks of age. When these mice were crossed with transgenic mice that overexpress human Bcl-2, there was a rescue of the facial motoneurons with a concomitant restoration of their normal soma size and expression of choline acetyltransferase. However, Bcl-2 overexpression did not prevent degeneration of myelinated axons in the facial and phrenic motor nerves and it did not increase the life span of the animals. Since Bcl-2 acts strictly on neuronal cell body survival without compensating for nerve degeneration in pmn/pmn/bcl-2 mice, this proto-oncogene would not in itself be sufficient for treatment of neurodegenerative diseases where axonal impairment is a major component. PMID- 7472524 TI - GAT-1, a high-affinity GABA plasma membrane transporter, is localized to neurons and astroglia in the cerebral cortex. AB - High affinity, GABA plasma membrane transporters influence the action of GABA, the main inhibitory neurotransmitter. The cellular expression of GAT-1, a prominent GABA transporter, has been investigated in the cerebral cortex of adult rats using in situ hybridization with 35S-labeled RNA probes and immunocytochemistry with affinity purified polyclonal antibodies directed to the C-terminus of rat GAT-1. GAT-1 mRNA was observed in numerous neurons and in some glial cells. Double-labeling experiments were performed to compare the pattern of GAT-1 mRNA containing and GAD67 immunoreactive cells. The majority of neurons expressing GAT-1 mRNA also contained GAD67 immunoreactivity (ir), but GAT-1 mRNA was also observed in a few pyramidal neurons. GAT-1-ir was localized to numerous puncta and fibers and to astrocytic processes, was not observed in sections incubated in GAT-1 antibodies preadsorbed with rat GAT-1 C-terminal peptide, and was observed in sections incubated in GAT-1 antibodies preadsorbed with the C terminal portion of the related peptides rat GAT-3(607-627) or rat glycine transporter-1(625-633). The highest number of GAT-1-ir puncta was in layer IV, followed by layers II-III. GAT-1 positive puncta appeared to have a preferential relationship to the soma and proximal dendrites of unlabeled pyramidal cells. All GAT-1 positive axon terminals formed symmetric synapses. This study demonstrates that (1) GAT-1 is expressed by both neurons and astrocytes, (2) the majority of GAT-1 expressing neurons contain GAD67, and (3) GAT-1 uptake system is more extensive than the GABA synthetizing system. These observations support the hypothesis that, in addition to its role in terminating GABA action by uptake into GABAergic axon terminals, GAT-1 influences both excitatory and inhibitory transmission by modulating the "paracrine" spread of GABA (Isaacson et al., 1993), and suggest that astrocytes may play an important role in this process. PMID- 7472525 TI - Observation and quantitation of exocytosis from the cell body of a fully developed neuron in Planorbis corneus. AB - We have discovered a neuronal system that releases neurotransmitter via exocytosis from the cell body. In the large dopamine cell of the pond snail Planorbis corneus, depolarization induces rhythmic release of dopamine from the cell body. When a stimulant is applied extracellularly or intracellularly in situ to the cell body, transient dopamine concentration packets that appear in a bursting pattern are observed. Dopamine release is calcium dependent and release is on the time scale expected for exocytosis (2 to 4 msec rise times). Quantitation of individual events reveals an average of 818,000 molecules per exocytotic event. As many as 89,000 individual exocytotic events have been observed following a single stimulation of one cell. Neurotransmitter exocytosis from the neuronal cell body appears to represent an alternative form of neurocommunication to synaptic transmission. PMID- 7472526 TI - [Introduction: the status and perspective of human immunodeficiency virus infection and its neurologic consequences]. PMID- 7472527 TI - [The epidemiology and cost of AIDS in France]. AB - Throughout the world the AIDS epidemic continues and even seems to accelerate in regions previously thought to be untouched. The number of cases has increased by 60% in one year. With its 15,000 seropositive persons and 35,000 cases of declared AIDS, France is the most affected of all European countries. Sexual contamination remains predominant with a relative increase of heterosexual transmissions. However, particularly in the South, transmission is prevalent in intra-venous drug addicts. In all cases worldwide it is the socially deprived, marginal and fragile populations which are the most vulnerable. Various previsions are possible in France, depending on the prophylactic measures applied. In any event, the number of seropositive subjects should be levelling out at the end of this decade, and the prevalence of the disease should stabilized at between 15,000 to 20,000 patients. At present, the cost of AIDS exceeds 5.5 billion francs. PMID- 7472528 TI - [Clinical and paraclinical diagnosis of AIDS neurologic lesions]. AB - Seventy to eighty percent of HIV-infected patients exhibit neurological disorders at an advanced stage of the disease. In almost 90% of cases anatomical examination of brains shows histological lesions. Even when often reversible neurological disorders occur during the HIV primary infection, most of the manifestations of central nervous system (CNS) damage remains the prerogative of severe immunodepression. The principal CNS lesions associated with HIV infection are presented here with the clinical and biological elements that lead to the diagnosis. Cerebral toxoplasmosis holds a privileged place in these manifestations since it responds to an efficient curative and prophylactic treatment with a well-codified medical care based on the test treatment. Biological data, therefore, only have a contributing value. HIV encephalopathy is frequent, but the dementia syndrome is less frequent than the finding of associated imaging and pathological anatomy: atrophy and lesions of the white matter. Thus, the dementia complex is an elimination diagnosis. Cryptococcosis must be systematically considered, not only in patients with meningeal symptoms and headaches, but also with those with isolated fever. The demonstration of cryptococcus and cryptococcic antigen in the CSF has an almost absolute diagnostic value; imaging plays a very small diagnostic role, looking for an exceptional cryptococcoma. Multifocal progressive leukoencephalopathy benefits from the accuracy of MRI, and the diagnosis is usually based on clinical data, MRI and evidence of the virus in the CSF by PCR, even though the only mean of obtaining full proof is, in theory, stereotaxic biopsy. Primary cerebral lymphoma is the diagnostic alternative to toxoplasmosis.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7472529 TI - [Early cerebral lesions in HIV infection. Postmortem radio-pathologic correlations in non-AIDS asymptomatic seropositive patients]. AB - In order to evaluate the diagnostic and prognostic value of MRI in the very early stages of HIV infection, we have compared the results of postmortem brain MRI and neuropathological studies in 7 asymptomatic HIV seropositive individuals, 8 seronegative controls with similar cause of death and 6 patients who died of AIDS in the absence of focal cerebral changes (opportunistic infection or tumour). Cerebral atrophy was consistently evaluated by both techniques. Seropositive asymptomatic cases were significantly more atrophic than the seronegative controls and significantly less atrophic than AIDS patients. Small high signal intensity areas in the white matter and basal ganglia were not significantly more frequent in seropositives than in seronegatives. No corresponding lesion was found at neuropathological examination. Diffuse myelin pallor of the cerebral white matter on myelin preparation was somewhat more severe in seropositive asymptomatic cases than in seronegative controls and less than in AIDS cases. However, these differences were not statistically significant. No significant correlation could be found between neuropathological myelin pallor and diffuse signal abnormalities of the white matter on MRI. We conclude that brain abnormalities are present at the early asymptomatic stage of HIV infection. These include vasculitis with opening of the blood brain barrier and consequent myelin pallor and gliosis of the white matter, and moderate brain atrophy. However MRI correlates are discrete or non specific on post mortem examination, and some probably correspond to scars of transient vascular inflammation. It is very unlikely that MRI examination has any diagnostic or prognostic value at the early stages of the disease. PMID- 7472530 TI - [HIV and dementia: neuropathology]. AB - Cognitive disorders associated with HIV infection may be due to focal lesions (lymphoma, toxoplasmosis, progressive multifocal leukoencephalitis, etc.), metabolic encephalopathy (e.g. hepatic insufficiency) or psychiatric disorders (depression). In the absence of such causes a "cognitive and motor syndrome associated with HIV infection" has been defined on clinical criteria (Working group of the American Academy of Neurology, 1991). This syndrome is not consistently associated with any specific lesion. Neither the multifocal encephalitis of HIV or CMV infection nor the diffuse leukoencephalopathy associated with HIV are the only causes. The existence of a neocortical neuronal loss has been suggested by several retrospective studies, but our prospective study has not shown cortical or subcortical atrophy. Measurement of neuronal density in Brodmann's areas 4,9 and 40 has not revealed a significant loss either global, by layer, or by column. The only constant lesion was gliosis of the cortex and white matter. Neuronal loss, therefore, is not indispensable to the occurrence of cognitive disorders in AIDS. The mechanism of dementia might be: dysfunction of cortical neurons (dendritic abnormalities, virus/neurotransmitter competition); subcortical dysfunction, as suggested by the high density of microglial nodules in that region; white matter lesions which could be due to abnormalities in the blood-brain barrier. The expression of cell adhesion molecules (VCAM-1, VLA-4, ICAM-1 and LFA-1) by endothelial cerebral cells is not significantly different in AIDS patients, demented or not, and in patients with multiple sclerosis. In contrast, the expression of VCAM-1 by astrocytes is significantly increased in demented AIDS patients compared with non demented ones.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7472531 TI - [Imagery of human immunodeficiency virus (HIV) encephalitis]. AB - The authors present the different aspects of HIV encephalitis in CT and MR in a series of 15 patients with anatomopathological proof. Atrophy was the most commonly found lesion (12 patients) and could rapidly be evolutive. White matter lesions (9 patients) were more of ten nodular than diffuse. They were better demonstrated by MRI and did not enhance after intra-venous injection of DOTA gadolinium. A frequent association with opportunistic infections (toxoplasmosis: 10 patients, CMV:2 patients), lymphoma (4 patients), or PML (3 patients) was observed which emphasized the low specificity of brain imaging in patients with HIV encephalitis. PMID- 7472532 TI - [AIDS dementia. Prognosis and therapeutic perspectives]. AB - AIDS dementia complex is a severe complication related to HIV infection of the central nervous system. It has been estimated to affect about 7% of AIDS patients, with an increased frequency in very young and old patients. In most cases, it appears in severely immuno-deficient patients. Typically, AIDS dementia presents as a subcortical dementia with cognitive, behavioural and motor decline. The mechanism of the dementia remains unclear; the neuronal loss is controversial and a neurotoxicity due to NMDA receptor activation is likely. No treatment proved to be efficient. Beneficial effect of anti-NMDA receptor antagonists are under investigation. PMID- 7472533 TI - [MRI aspects of progressive multifocal leukoencephalopathy]. AB - Progressive Multifocal Leukoencephalopathy is a demyelinating disease. MRI shows high signal intensity areas on T2w sequence and low intensity aeras on T1w sequence, without enhancement after intravenous contrast injection. The involvement of arcuate fiber (U fibers) creates a sharp border with the cortex. There is no mass effect. Involvement of parieto-occipital areas is frequent. The lesions may be uni or bilateral, single or multiple; bilateral lesions are asymmetric. This typical appearance on MR images occurs in 90% of the patients with PML. Some atypical patterns may occur: focal hemorrhage, atrophy, faint peripheral enhancement and involvement of deep gray matter (basal ganglia). In most cases, the clinical and MR features provide the diagnosis. The main differential diagnosis, in MRI, is HIV-leukoencephalitis, but lesions are diffuse, less intense on T2w sequence and not visible on T1, without involvement of the arcuate fibers. Stereotactic biopsy should be performed only for atypical lesions, particularly in case of predominant involvement of deep gray structures. PMID- 7472534 TI - [Pathologic anatomy of cytomegalovirus encephalomyelitis and varicella-zona virus encephalomyelitis]. AB - Cytomegalovirus (CMV) infection of the nervous system is frequent in acquired immunodeficiency syndrome (AIDS) and can be responsible for encephalitis, encephalomyelitis, meningoradiculitis or polyradiculo-neuropathy. Encephalitis is characterized at microscopy by its periventricular and cerebellar location, and by the presence of cytomegalic cells, containing intranuclear and/or intracytoplasmic inclusions, microglial nodules and necrotic foci. The virus can infect almost all types of cells. Coexistence of CMV and HIV has been observed in giant cells of macrophagic origin. It has been suggested that the two viruses could act in synergy. The nervous system is seldom infected by the varicella zoster virus (VZV) in AIDS. The infection can be responsible for multifocal leukoencephalitis, ventriculitis, vascular lesions associated or not with cerebral infarction, or with meningomyeloradiculitis. In almost all cell types Cowdry's type A intranuclear inclusions have been found. The virus can be demonstrated by immunohistochemistry or in situ hybridization. VZV antigens have been reported in the walls of vessels damaged by a non inflammatory obliterating vasculopathy or by a granulomatous angiitis. Coexistence of VZV and HIV has been observed in giant cells of macrophagic origin, and synergy between those two viruses has been suspected. PMID- 7472535 TI - [CMV and VZV encephalitis in AIDS]. AB - MATERIAL AND METHODS: Eighty patients were followed up prospectively. Histological correlation was obtained in 25 cases. All MRI examinations were performed on at 0.5 Tesla in T1-weighted sequences with and without gadolinium injection, and in axial or frontal T2-weighted spin echo sequences. Histological confirmation was obtained 30 days on average after the last MRI examination. Immunohistochemical stainings were performed in search of CMV, VZV, toxoplasma, HIV antigen and lymphoma. RESULTS: CMV meningoencephalitis was found in 6 cases. In 3 of these it was manifested by atrophy, either isolated or associated with high signal intensity punctiform areas. Histology detected cortical or subcortical microglial nodules. In 2 cases MRI displayed abnormalities of subependymal nodular signals without enhancement, associated with punctiform abnormalities of subcortical signals. Histology showed subependymal foci of necrosis and abnormalities of white matter. In one case, MRI showed a ventriculitis confirmed by histology. VZV meningoencephalitis was diagnosed in 2 cases. MRI displayed abnormal basal ganglia related to meningitis (n = 1). All abnormalities were confirmed at histology. CONCLUSION: Some images (ventriculitis, infarction in basal ganglia, abnormal subependymal signal) would suggest VZV and CMV encephalitis. Other images (abnormalities of punctiform signals or atrophy) are not specific. PMID- 7472536 TI - [The impact of the prevention of cerebral toxoplasmosis]. AB - In France, where 70% of adults are latently infected by toxoplasma, from 20% to 40% of patients with AIDS developed toxoplasmic encephalitis until recently. The prophylactic use of drugs which are active against pneumocystis and toxoplasma has proven to be efficient. These drugs are trimethoprim-sulfamethoxazole or dapsone-pyrimethamine. With the extent of these primary prophylaxis, there is a decrease of risk of toxoplasma encephalitis; thus the rate of toxoplasma encephalitis among opportunistic infections has fallen off from 19% of the patients in 1988 to 6% in 1994, in the department of infectious diseases of the Pitie-Salpetriere hospital. However, toxoplasmic abscesses occurring despite the prophylaxis are frequently slow growing lesions which can become huge with a moderate mass effect, mimicking the pattern of primary cerebral lymphoma. The rule of antitoxoplasmic trial treatment must be strictly followed, even under prophylaxis. PMID- 7472537 TI - [Cerebral toxoplasmosis. Neuroradiologic diagnosis and prognostic monitoring]. AB - Cerebral toxoplasmosis remains the most frequent etiology of cerebral masses in AIDS. In most cases, the disclosure of multiple enhanced masses is suggestive of diagnosis of toxoplasmosis and leads to undertake presumptive therapy. Sometimes, the pattern is less suggestive, and the possibility of primary cerebral lymphoma (PCL) is a diagnostic dilemma, because this is a short term life-threatening lesion in the absence of an undelayed fitting therapy. However, apart from the periventricular form of PCL, where directly proceeding to biopsy should be suggested, there is no distinctive pattern of PCL. The most reliable features in distinguishing between PCL and toxoplasmosis are: solitary lesion, homogeneous enhancement of a lesion whose diameter is larger than 2 cm, situation in deep periventricular white matter and limited edema and mass effect. The presence of haemorrhagic foci in the lesion, if there is no steroid therapy is rather indicative of toxoplasmosis. The lack of enhancement on enhanced CT-scan does not allow to eliminate cerebral toxoplasmosis, and indicate MR examination, or empiric antitoxoplasmic treatment. The radiologic follow-up depends on clinical response and on initial pattern. The criteria of response to antitoxoplasmic treatment are: the decrease in volume or number of lesions, the decrease of edematous reaction and mass effect (if no steroid was administered). If the lesion was initially equivocal, the lack of growth in a 8-day delay is a criterion for toxoplasmosis, just as the occurring of haemorrhage in the lesions (if there is no steroid-therapy). PMID- 7472538 TI - [The epidemiology of cerebral lymphoma in AIDS]. AB - Cerebral lymphomas are one of the complications of AIDS. Their increasing frequency, especially in the last decade, has to be correlated with the incidence of systemic lymphomas and especially central nervous system (CNS) lymphomas in non immuno-compromised patients. CNS involvement is found in 40% of systemic lymphomas. Primary lymphomas have an incidence of 0.4-0.56% in epidemiological studies and 7.6% in post-mortem series. Primary lymphomas are noted in all patients, homosexuals, drug-addicts, children, with a lower rate in the haemophiliacs. Known risk factors are HIV itself Epstein-Barr virus almost constantly demonstrated in tumour cells. Epidemiological studies on cerebral lymphomas must still be completed as, until recently, they were included in the group entitled lymphomas, and risk factors have to be specified with more accuracy. PMID- 7472539 TI - [The diagnosis of primary cerebral lymphoma in AIDS. The contribution of imaging]. AB - Apart from the very frequent HIV encephalitis, which lays the foundation for opportunistic infections, the most common diseases encountered in HIV-infected patients are toxoplasmosis and lymphoma; the percentage of cases of other diseases is very small. It is capital to diagnose cerebral lymphoma at an early stage in these patients who already are in a precarious general and neurological state since this type of lesion usually occurs late in the natural course of AIDS. In the differential diagnosis between toxoplasmosis and lymphoma only stereotaxic biopsy enables a positive diagnosis to be made, but imaging methods, such as CT and MRI, provide data that help guide to a diagnosis which sometimes may be definitive. Suggestive of lymphoma is a single infiltrating lesion wider than 4 cm, which is paraventricular or located in the posterior fossa, has little perilesional alteration and a short tumoral doubling time during the imaging follow-up. Suggestive of toxoplasmosis are multiple, small, annular or nodular lesions with an important perilesional inflammation and usually located in basal ganglia. In addition, in MRI the kinetics of enhancement after contrast injection is intense and occurs early in lymphoma, in contrast with the toxoplasmic abscesses, and this should provide a more specific differential diagnosis. Scintigraphic studies with somatostatin or positron emission tomography, using fluorodeoxyglucose (FDG-PET scintigraphy), also seem to be an interesting mean of making a specific diagnosis of cerebral lesion, according to a principle that is close to dynamic MRI. In lymphoma, capturing of the tracer is about 3 times greater than in infective lesions, notably the toxoplasmic ones. Imaging, therefore, is provided with tools which permit an increasingly specific approach to the primary cerebral lymphoma of AIDS, the definitive diagnosis of which rests on stereotaxic biopsy. This high specificity facilitates a better selection of patients requiring this procedure and shortens the delay in its execution. PMID- 7472540 TI - [Cerebral lesions in AIDS: what can be expected from scintigraphy? Cerebral tomographic scintigraphy using thallium-201: a contribution to the differential diagnosis of lymphomas and infectious lesions]. AB - CT and MRI are not consistently able to differentiate central nervous system (CNS) lymphoma from focal cerebral infection such as toxoplasmosis in the human immunodeficiency virus (HIV) involvement. In this study thallium 201 (and/or technetium-MIBI) SPECT was performed in 6 patients suffering from AIDS and CNS lesions. An index based on the ratio of thallium uptake in the lesion vs the contralateral scalp was calculated. In 4 out of 5 patients with lymphoma (3 confirmed by biopsy, 2 highly suspected on CT and resistance to antitoxoplasmosis treatment) focal lesions showed high uptake of thallium. On the other hand one markedly necrotic lymphoma and all infectious lesions did not take up thallium. This suggests a role for thallium 201 brain SPECT in the workup of focal CNS lesions in AIDS. PMID- 7472541 TI - Responding to change: the challenge of the 1990s. AB - In 1995, neurosurgery in the United States is facing challenges and threats never before imagined. In its attempts to respond as a relatively small specialty, the neurosurgical community can no longer afford to remain a house divided. Neurosurgical unit has now become a critical matter. In particular, our neurosurgical leadership must minimize duplication of efforts and efficiently allocate its resources. Most importantly, it must unify its energies in directing the numerous joint American Association of Neurological Surgeons/Congress of Neurological Surgeons activities, as well as serving as a direct liaison with the emerging representative assembly under the Joint Council of State Neurosurgical Societies. The unification of the neurosurgical leadership will clearly enhance our ability to deal with all of our present challenges and future changes, but more importantly it will permit our specialty to truly speak with one voice. PMID- 7472542 TI - Long-term prognosis and effect of endarterectomy in patients with symptomatic severe carotid stenosis and contralateral carotid stenosis or occlusion: results from NASCET. North American Symptomatic Carotid Endarterectomy Trial (NASCET) Group. AB - The purpose of this study was to examine how the prognosis of patients who presented with a recent ischemic event referable to a 70% to 99% stenosis of one carotid artery (ipsilateral) was altered by stenosis and occlusion of the contralateral carotid artery. The benefit of performing carotid endarterectomy on the recently symptomatic artery, in the presence of contralateral artery disease, was also examined. A total of 659 patients were grouped into one of three categories according to the extent of stenosis in the contralateral carotid artery: less than 70% (559 patients), 70% to 99% (57 patients), and occlusion (43 patients). Strokes that occurred during the follow-up period were designated as ipsilateral if they arose from the same carotid artery as the symptom for which the patient had been entered into the study. Medically treated patients with an occluded contralateral artery were more than twice as likely to have had an ipsilateral stroke at 2 years than patients with either severe (hazard ratio: 2.36; 95% confidence interval (CI): 1.00-5.62) or mild-to-moderate (hazard ratio: 2.65; 95% CI: 1.43-4.90) contralateral artery stenosis. The perioperative risk of stroke and death was higher in patients with an occluded contralateral artery (4.0% risk) or mild-to-moderate (5.1% risk) contralateral stenosis. Regression analyses indicated that the results were not affected by other risk factors. An occluded contralateral carotid artery significantly increased the risk of stroke associated with a severely stenosed ipsilateral carotid artery. Despite higher perioperative morbidity in the presence of an occluded contralateral artery, the longer-term outlook for patients who had endarterectomy performed on the recently symptomatic, severely stenosed ipsilateral carotid artery was considerably better than for medically treated patients. PMID- 7472543 TI - Surgical outcome for intra- and extrapelvic femoral nerve lesions. AB - Seventy-eight traumatic neuropathies were seen in 94 patients with femoral nerve lesions; 54 of these were operated on because of persistent complete functional loss and/or pain. The most common mechanism of injury to the femoral nerve was iatrogenic due to inguinal herniorrhaphy, total hip replacement, intraabdominal vascular or gynecological operation, and, less commonly, appendectomy, lumbar sympathectomy, and laparoscopic procedures. Femoral nerve injuries also resulted from penetrating gunshot and stab wounds, laceration by glass, and stretch/contusive injuries associated with pelvic fractures. There were no signs of clinical or electrical recovery in 45 of 78 patients with traumatic nerve injuries. These and other partial injuries associated with pain were explored and evaluated by intraoperative nerve stimulation and recording of nerve action potentials (NAPs). Despite complete loss of nerve function preoperatively, 13 patients had recordable NAPs and underwent neurolysis; each recovered function to at least a Grade 3 level. Twenty-seven patients had sural graft repairs performed with graft lengths varying from 2.5 to 14 cm. Most patients had some nerve regeneration and regained function to Grade 3 to 4 levels by 2 years postoperatively. Four of five patients with suture repairs recovered to Grade 3 or better within 2 years postoperatively. Despite a proximal pelvic level for most of these injuries and, as a result, lengthy graft repairs, recovery of some useful function was the rule rather than the exception. Tumors involved the femoral nerve in 16 patients and included eight neurofibromas, four schwannomas, one neurogenic sarcoma, two ganglion cysts, and one leiomyosarcoma. All tumors were treated surgically and most were removed successfully. PMID- 7472544 TI - Prolonged external ventricular drainage with percutaneous long-tunnel ventriculostomies. AB - External ventricular drainage has been used extensively for management of several neurosurgical disorders. The main limitation of this procedure has been the high risk of infection, especially with prolonged drainage. In an effort to minimize the risk of infection, the authors have used a new ventriculostomy technique that involves tunneling the ventricular catheter subcutaneously to an exit site in the lower chest or upper abdomen. This report describes the results of this procedure on 100 consecutive cases. Patients requiring emergency ventriculostomies had short-tunnel ventriculostomies placed at the bedside that were converted to long tunnel ventriculostomies in the operating room within 5 days. The average duration of drainage was 18.3 days (range 5-40 days). Cerebrospinal fluid was routinely sent for Gram staining and culture to monitor for infection. Prophylactic antibiotic medications were administered only perioperatively. No infection was observed during the first 16 days of drainage in any patient. The overall incidence of infection was 4% and blockage occurred in 6% of the cases. In this series the incidence of ventricular infection was 2.37 per 1000 ventricular drainage days, one of the lowest reported incidences of infection in the literature. This procedure provides a simple and effective method of maintaining long-term ventricular drainage with a very low risk of infection or blockage. PMID- 7472545 TI - Localization of inactive cerebrospinal fluid fistulas. AB - Because of the importance of preoperative localization of dural fistulas, many imaging modalities have been critically evaluated for their role in pinpointing the site of cerebrospinal fluid (CSF) leakage. Twenty-one consecutive patients who were suspected of having a CSF fistula were studied to evaluate magnetic resonance (MR) imaging in locating the fistula. These patients were also studied independently by fine-slice computerized tomography (CT). The MR images demonstrated lesions compatible with dural fistulas in 19 patients, whereas CT demonstrated only seven of these lesions. All of these patients underwent surgical dural repair. The remaining two patients underwent surgical exploration on the basis of the CT findings but no dural fistula was found in either patient. All patients made a good postoperative recovery. One patient developed a postoperative wound infection and in another CSF leakage recurred. Although MR imaging was very precise in locating the CSF fistulas, CT missed a significant number of these lesions and was falsely positive in 9.5% of cases. Therefore, it is concluded that MR imaging is an essential investigation in patients with a suspected dural fistula and should be performed before embarking upon surgery and before assuming natural healing of the CSF fistula. PMID- 7472546 TI - Demonstration of neurovascular compression in trigeminal neuralgia with magnetic resonance imaging. Comparison with surgical findings in 52 consecutive operative cases. AB - Until recently, the inability to demonstrate neurovascular compression of the trigeminal nerve preoperatively resulted in surgery being offered only in cases of severe trigeminal neuralgia (TGN), frequently after a prolonged trial of medical treatment and following less invasive procedures, despite the fact that posterior fossa microvascular decompression gives long-term pain relief in 80% to 90% of cases. To assess whether vascular compression of the nerve could be demonstrated preoperatively, high definition magnetic resonance tomographic angiography (MRTA) was performed in 50 consecutive patients, five of whom had bilateral TGN, prior to posterior fossa surgery. The imaging results were compared with the operative findings in all patients, including two patients who underwent bilateral exploration. Vascular compression of the trigeminal nerve was identified in 42 of 45 patients with unilateral symptoms and on both sides in four patients with bilateral TGN. In the last patient with bilateral TGN, neurovascular compression was identified on one side, and on the other side the compressing superior cerebellar artery was separated from the nerve by a sponge placed during previous surgery. There was full agreement regarding the presence or absence of neurovascular compression demonstrated by MRTA in 50 of 52 explorations, but MRTA misclassified four vessels compressing the trigeminal nerve as arteries rather than veins. In two cases, there was disagreement between the surgical and MRTA findings. In the first of these cases, surgery revealed distortion of the nerve at the pons by a vein that MRTA had predicted to lie 6 mm remote from this point. In the second patient, venous compression was missed; however, this patient was investigated early in the series and did not have gadolinium-enhanced imaging. In nine cases, MRTA correctly identified neurovascular compression of the trigeminal nerve by two arteries. Moreover, MRTA successfully guided surgical reexploration in one patient in whom a compressing vessel was missed during earlier surgery and also prompted exploration of the posterior fossa in two patients with multiple sclerosis and one patient with Charcot-Marie-Tooth syndrome, in whom neurovascular compression was identified preoperatively. It is concluded that MRTA is an extremely sensitive and specific method for demonstrating vascular compression in TGN. As a result, open surgical procedures can be recommended with confidence, and microvascular decompression is now the treatment of choice for TGN at the authors' unit. They propose MRTA as the definitive investigation in such patients in whom surgery is contemplated. PMID- 7472547 TI - Late patency of long saphenous vein bypass grafts to the anterior and posterior cerebral circulation. AB - To evaluate the late results and the natural history of long saphenous vein bypass grafts (SVGs) between the extracranial and intracranial circulation, the authors retrospectively analyzed 202 consecutive SVGs performed at the Mayo Clinic from 1979 to 1992. The distal anastomosis was to the vertebrobasilar system in 98 patients and to the carotid artery system in 103 patients. Surgical indications were advanced cerebroocclusive disease in 63% (127 cases), giant aneurysm in 37% (74 cases), and neoplasm in one patient. In 125 patent SVGs follow-up information was obtained for longer than 1 year and in 23 patent SVGs it was over 10 years (maximum 13 years, median 6.5 years). Most of the graft failures (76%) occurred during the 1st year after surgery, with 42% of all graft failures found during the first 24 hours after operation. Late graft attrition occurred in only 10 patients (8%). Cumulative patency at 1 year was 86% +/- 3%, at 5 years 82% +/- 4%, and at 13 years 73% +/- 19%. Neurological worsening at the time of occlusion developed in 72% of patients with early occlusion, whereas 80% of patients with late graft occlusion had no new neurological symptoms. Long-term patency of SVGs for cerebral revascularization appears to be excellent, with an average failure rate of 1% to 1.5% per year following the 1st year after surgery. To minimize early graft thrombosis, meticulous attention must be paid to technical detail. PMID- 7472548 TI - Cerebral arterial aneurysm formation and rupture in 20,767 elderly patients: hypertension and other risk factors. AB - Cerebral arterial aneurysms are common in the general population and their rupture is a catastrophic event. Considerable uncertainty remains concerning the conditions that predispose individuals to aneurysm formation or rupture. The role of systemic hypertension in aneurysm formation and rupture has been especially controversial. Demographic variables have rarely been addressed because of the small sample sizes in previous studies. The authors describe the demographics and prevalence of hypertension in 20,767 Medicare patients with an unruptured aneurysm and compared these to a random sample of the hospitalized Medicare population. The prevalence of hypertension in patients with unruptured aneurysms was 43.2% compared with 34.4% in the random sample. Patients who survived their initial hospitalization were separated into two groups: those with an unruptured cerebral aneurysm as the primary diagnosis and those with an unruptured cerebral aneurysm as a secondary diagnosis. Follow-up data for 18,119 patients were examined to determine the risk of subarachnoid hemorrhage (SAH) associated with age, gender, race, hypertension, insulin-dependent diabetes mellitus, and surgical treatment. For patients with an unruptured cerebral aneurysm as the primary diagnosis, hypertension was found to be a significant risk factor for future SAH (risk ratio: 1.46, 95% confidence interval (CI): 1.01-2.11), whereas surgical treatment (risk ratio: 0.29, 95% CI: 0.09-0.97) had a significant protective effect. Advancing age had a small but significant protective effect in both groups. Elderly patients identified with unruptured aneurysms are more likely to have coexisting hypertension than the general hospitalized population. In elderly patients hospitalized with an unruptured cerebral aneurysm as their primary diagnosis, hypertension is a risk factor for subsequent SAH, whereas surgical treatment is a protective factor against SAH. PMID- 7472549 TI - The natural history of cerebral cavernous malformations. AB - To determine the natural history of brain cavernous malformations, the authors entered patients referred to their center into a prospective registry between 1987 and 1993. All patients underwent magnetic resonance imaging, which showed the typical appearance of this lesion, and conservative management was recommended in all. Patients or their referring physicians were contacted for follow-up data. The purpose of the study was to define the rate of symptomatic hemorrhage and to determine the outcome in those patients who had suffered seizures. Follow-up data were available for 122 patients with a mean age at entry of 37 years (range 4-82 years). The malformation was located in the brainstem in 43 cases (35%), the basal ganglia/thalamus in 20 (17%), and a hemispheric area in 59 (48%). Fifty percent of patients had never had a symptomatic hemorrhage, 41% had one bleed, 7% had two, and 2% had three. Seizures were reported in 23% of patients and headaches in 15%. Lesions were solitary in 80% of patients and multiple in 20%. The retrospective annual hemorrhage rate (61 bleeds/4550.6 patient-years of life) was 1.3%. The mean prospective follow-up period was 34 months. There were nine bleeds during this time, six with new neurological deficits. In patients without a prior bleed, the prospective annual rate of hemorrhage was 0.6%. In contrast, patients with prior hemorrhage had an annual bleed rate of 4.5% (p = 0.028). Patient sex (p = 0.97) or the presence of seizures (p = 0.11), headaches (p = 0.06), or solitary versus multiple lesions (p = 0.15) were not significant predictors of later hemorrhage. There was no difference in the rate of bleeds between brain locations. Four patients with seizures became seizure-free and four patients without seizures later developed seizures; only one patient developed intractable seizures. Fourteen had radiosurgery. No patient died in the follow-up period. This study indicates that conservative versus operative management strategies may need to be redefined, especially in patients who present with hemorrhage and who appear to have a significantly increased risk of subsequent rehemorrhage. PMID- 7472550 TI - Reduction of hemorrhage risk after stereotactic radiosurgery for cavernous malformations. AB - The benefits of radiosurgery for cavernous malformations are difficult to assess because of the unclear natural history of this vascular lesion, the inability to image malformation vessels, and the lack of an imaging technique that defines "cure." The authors selected for radiosurgery 47 patients who harbored a hemorrhagic malformation in a critical intraparenchymal location remote from a pial or ependymal surface. Of these, 44 patients had experienced at least two hemorrhages before radiosurgery. The mean patient age was 39 years; six patients had previously undergone attempted surgical removal. The malformation was located in the pons/midbrain in 24 cases, the medulla in three, the thalamus in nine, the basal ganglia in three, deep in a parietal lobe in four, and deep in a temporal lobe in four. Patients had sustained initial hemorrhages from 0.5 to 12 years prior to radiosurgery (mean 4.12 years). In these patients, who were not typical of the majority of patients with cavernous malformations, there were 109 bleeds before radiosurgery in 193 prior observation-years, for 56.5% annual hemorrhage rate (including the first hemorrhage), or an annual rate of 32% subsequent to the first hemorrhage. The mean follow-up period after radiosurgery was 3.6 years (range 0.33-6.4 years). The proportion of patients with hemorrhage after radiosurgery was significantly reduced (p < 0.0001), as was the mean number of hemorrhages per patient (p = 0.00004). In the first 2 years after radiosurgery, there were seven bleeds in 80 observation-years (8.8% annual hemorrhage rate). In the 2- to 6-year interval after radiosurgery, the annual rate decreased to 1.1% (one bleed). After radiosurgery, 12 patients (26%) sustained neurological worsening that correlated with imaging changes. In eight patients these deficits were temporary; two underwent surgical resection and died. Two patients had new permanent deficits (4%). A significant reduction was observed in the hemorrhage rate after radiosurgery in patients who had deep hemorrhagic cavernous malformations, especially after a 2-year latency interval. This evidence provides further support to the belief that radiosurgery is an effective strategy for cavernous malformations, especially when located within the parenchyma of the brainstem or diencephalon. PMID- 7472551 TI - Interim report on the radiosurgical treatment of cerebral arteriovenous malformations. The influence of size, dose, time, and technical factors on obliteration rate. AB - During the authors' initial 4-year experience with radiosurgery using the Leksell cobalt-60 gamma unit, they treated 121 patients with cerebral arteriovenous malformations (AVMs). The radiosurgical dose to the margin of the nidus was 20 Gy for lesions less than 2.0 cm in diameter (volume < or = 4.2 cm3); 18 Gy for malformations 2.1 to 3.0 cm in diameter (volume 4.2-14.1 cm3); and 16 Gy for malformations greater than 3.0 cm (volume > 14.1 cm3). Fifty-one patients underwent follow-up angiography between 1 and 3 years after treatment, and complete obliteration of the nidus was confirmed in 38 (74.5%) of these patients. Thirty-two (74.4%) of 43 AVMs with volumes of 10 cm3 or less and six (75%) of eight larger AVMs (volume 11-30 cm3) showed complete obliteration. Analysis of the time course of AVM nidus shrinkage and obliteration showed that most of the radiosurgically induced effect had occurred by 36 months after treatment. Retrospective analysis of the dose plans for 10 AVMs that were not obliterated by 36 months after gamma knife radiosurgery at the authors' institution (eight cases) or elsewhere (two cases) revealed that six AVMs had not been covered completely by the prescribed isodose. Six (5%) of the 121 patients developed neurological deficits as a direct result of radiosurgical treatment. The authors infer from these data that malformations up to 30 cm3 in volume (approximately 4.0 cm in average diameter) can be treated effectively with an acceptably low complication rate using a radiosurgical dose of 16 Gy to the margin of the nidus. The obliteration rate for the larger malformations that were treated with a dose of 16 to 18 Gy appears to be similar to that for smaller ones treated with 18 to 20 Gy. As more experience accrues using radiosurgery to treat AVMs, patient selection criteria and the variables associated with successful obliteration of the nidus should become more clearly defined. PMID- 7472552 TI - Treatment of carotid-cavernous sinus fistulas using a superior ophthalmic vein approach. AB - The authors describe the method and results of treatment of 12 consecutive patients with carotid-cavernous sinus fistulas (CCFs). Treatment was by embolization via a transvenous approach through the superior ophthalmic vein (SOV). The CCFs (two direct and 10 dural) had previously been treated unsuccessfully or, for mechanical reasons, could not be treated by the standard techniques of endoarterial balloon occlusion, particle or glue embolization of feeding vessels from one or both external carotid arteries, or transvenous occlusion of the fistula via the ipsilateral inferior petrosal sinus. All 12 patients were successfully treated either by advancement of a detachable balloon catheter through the ipsilateral SOV into the cavernous sinus with subsequent inflation and detachment of the balloon (11 patients) or by introduction of multiple thrombogenic coils into the fistula via the ipsilateral SOV (one patient). All patients had complete resolution of symptoms and signs after successful occlusion of the CCF. There were no intraoperative complications; however, one patient required postoperative embolization of a residual posteriorly draining fistula via the ipsilateral external carotid artery, and another developed a persistent abducens nerve paresis that eventually required surgical correction. Ten (83.3%) of the 12 patients underwent cerebral angiography 3 to 6 months after surgery, and none showed evidence of a recurrent fistula. Similarly, none of the 12 patients developed recurrent symptoms and signs suggesting recurrence of the fistula during a follow-up period that ranged from 6 months to 10 years (mean 64 months). It is concluded that the transvenous approach to the cavernous sinus through the SOV is a safe and effective treatment of both direct and dural CCFs that are not amenable to transarterial or other transvenous approaches. PMID- 7472553 TI - Neurosurgical management of cerebral aneurysms following unsuccessful or incomplete endovascular embolization. AB - Modern endovascular techniques permit treatment of intracranial aneurysms in many circumstances when surgery is associated with significant morbidity. Occasionally, embolization of aneurysms is unsuccessful or incomplete or followed by complications, in which case surgical management is required. Since 1986, 196 patients have undergone embolization of intracranial aneurysms at the authors' institution and 21 (11%) required subsequent surgical treatment. Attempted embolization failed in five patients (Group A). Ten patients (Group B) had only partial occlusion of the aneurysm or demonstrated recanalization on follow-up studies. Eight of these Group B patients underwent embolization with Guglielmi detachable coils (GDCs), representing 5.7% of the 141 GDC-treated patients in this experience. Surgical treatment in these two groups consisted of clipping (eight cases), surgical parent vessel occlusion (one case), and parent vessel occlusion with extracranial-intracranial bypass (six cases). Fourteen (93%) of the 15 patients in these two groups had an excellent or good outcome with complete aneurysm occlusion. Six patients underwent surgery to treat complications related to the endovascular procedure (Group C). Of these, four patients had neurological improvement compared to their preoperative state, and two died. This series of cases demonstrates that surgical treatment of aneurysms is usually possible with good results following incomplete embolization and emphasizes the need for close and continued neurosurgical involvement in the endovascular management of intracranial aneurysms. PMID- 7472554 TI - Functional MR and PET imaging of rolandic and visual cortices for neurosurgical planning. AB - Magnetic resonance (MR) imaging has recently been used to demonstrate physiological activation of the human brain. This development is of considerable interest to the neurosurgeon planning procedures near brain regions involving specific functions. In the present study, rolandic and visual cortices were imaged with a conventional 1.5-tesla clinical MR imager using a spoiled gradient recalled acquisition in the steady state sequence. Two patients, one with a right frontal astrocytoma and the other with a left parietal meningioma, underwent MR imaging of rolandic cortex while performing a repetitive finger apposition task. Two patients with complex partial seizures referable to the temporal and occipital regions underwent MR imaging of visual cortex while exposed to repetitive photic stimulation (8.3 Hz). Significant signal intensity changes up to 15% between the activation and rest conditions were observed near the surgical targets at the expected anatomical location of the rolandic and visual cortices. In two of these cases activation measured by MR was compared and found similar to the activation measured at the same plane by H2(15)O positron emission tomography (PET). These results suggest that functional MR and PET techniques can be used to obtain preoperative brain mapping in individual patients considered for neurosurgical procedures. PMID- 7472555 TI - Hemin activation of an inducible isoform of nitric oxide synthase in vascular smooth-muscle cells. AB - Hemin is a prominent breakdown product of hemoglobin, and high levels of hemin are found in the cerebrospinal fluid during subarachnoid hemorrhage-induced vasospasm. The possible role of hemin in modifying vascular function was examined in the present study by testing its effects on nitric oxide synthase (NOS) activity in cultured rat aortic smooth-muscle cells. Nitric oxide synthase activity was estimated from the amounts of accumulated nitrite and nitrate, which are oxidative products of nitric oxide (NO). Hemin (1-100 microM) increased the levels of nitrite and nitrate in culture medium in a dose- and time-dependent manner. The hemin-induced elevation of nitrite and nitrate was inhibited significantly by the NOS inhibitor, N omega-nitro-L-arginine (300 microM), and by the protein synthesis inhibitor, cycloheximide (5 micrograms/ml). These results indicate that hemin is capable of stimulating the expression of an inducible isoform of NOS (iNOS) in vascular smooth muscle. Transcriptional expression of iNOS is known to cause injurious effects on the maintenance of cellular homeostasis by generating extremely high levels of NO. The generation of hemin from methemoglobin during hemolysis of a subarachnoid blood clot could therefore stimulate an excessive production of NO in vascular smooth-muscle cells. It is postulated that this series of events contributes to the development of vascular injury associated with cerebral vasospasm after aneurysmal subarachnoid hemorrhage. PMID- 7472556 TI - Functional changes in cultured strips of canine cerebral arteries after prolonged exposure to oxyhemoglobin. AB - The present study was undertaken to determine whether oxyhemoglobin (OxyHb) is responsible for the functional alterations in the cerebral arteries observed during chronic vasospasm after subarachnoid hemorrhage. Vascular strips of canine basilar arteries were kept in organ culture for 3 days with or without repetitive exposure to OxyHb (OxyHb-treated and control strips). Contractions elicited by high levels of potassium (80 mM) and uridine 5'-triphosphate (3 x 10(-4) M) were reduced in the OxyHb-treated group in a concentration-dependent manner. The relaxations evoked by nitric oxide and 8-bromo-cyclic guanosine monophosphate (8 bromo-cGMP) were not affected. Relaxations elicited by the calcium channel blocker, diltiazem, were attenuated in the OxyHb-treated rings. When the extracellular calcium concentration ([Ca2+]e) was changed from a concentration in the external solution of 10(-8) M to 10(-3) M, myogenic tension developed. Myogenic tension, expressed as a percentage of the maximum contraction in each segment, was augmented in the OxyHb-treated group at [Ca2+]e of 10(-5) M and 10( 4) M. There were no significant differences in passive compliance of the arterial wall between the two groups. These results demonstrated that prolonged exposure to OxyHb in vitro results in a decrease in contractile capacity and an increase in sensitivity to [Ca2+]e, in agreement with previous findings in spastic arteries. By contrast, impairment of the 8-bromo-cGMP-mediated relaxation pathway and increased stiffness of the arterial wall, which have been reported to occur in spastic arteries, were not induced by prolonged exposure to OxyHb in vitro. PMID- 7472557 TI - Anesthetic-dependent pial arteriolar response to ethanol. AB - Anesthetic agents are often administered in the presence of ethyl alcohol, both in research and in the clinical setting. The authors tested the hypothesis that anesthetic agents may affect cerebrovascular responses to ethanol. A closed cranial window preparation in the rat was used to compare the response of pial arterioles to topically applied ethanol (0.01% to 1% vol/vol) in the presence of alpha-chloralose/urethane (50 and 600 mg/kg, respectively) or halothane (0.5% to 1%) anesthesia. Heart rate, mean arterial blood pressure, and blood gas levels were maintained stable and within the physiological range throughout each experiment. Ethanol induced significant vasoconstriction in alpha chloralose/urethane-anesthetized animals (multivariate analysis of variance (MANOVA), p = 0.039); conversely, ethanol induced significant vasodilation of the pial arterioles in halothane-anesthetized animals (MANOVA, p = 0.017). These responses were significantly different from one another (MANOVA, p = 0.001). Thus, the choice of anesthetic agent alters the cerebrovascular response to ethanol, and care should be taken to ascertain the influence of anesthesia in both research and clinical settings. PMID- 7472558 TI - Chronic postinjury administration of MDL 26,479 (Suritozole), a negative modulator at the GABAA receptor, and cognitive impairment in rats following traumatic brain injury. AB - The present experiment examined the efficacy of postinjury administration of MDL 26,479 (Suritozole), a negative modulator at the gamma-aminobutyric acidA (GABAA) receptor that enhances cholinergic function, in attenuating spatial memory deficits after traumatic brain injury in the rat. Two experiments were performed. In the delayed-dosing experiment, rats received a moderate level (2.1 atm) of fluid-percussion brain injury and were tested in the Morris water maze 11 to 15 days following injury. These rats were injected with either 5 mg/kg (eight rats) or 10 mg/kg (eight rats) of MDL 26,479 60 minutes before each water maze test. Additional rats were injured and treated with saline (eight rats) or were surgically prepared but not injured (eight rats). In the second experiment, an early postinjury dosing procedure was followed. Rats were injured in the same manner but drug treatment began 24 hours after injury and continued daily through Day 15. Results indicated that the rats in the delayed chronic dosing regimen did not differ from the injured, saline-treated rats in their latency to reach the goal platform (p > 0.05). However, those treated chronically beginning 24 hours after injury had significantly shorter latencies than the injured, saline-treated rats (p < 0.05). These results suggest that administration of agents that enhance cholinergic function may be an appropriate strategy for promoting cognitive recovery when given after traumatic brain injury. Furthermore, prolonged treatment may be necessary to elicit beneficial effects. PMID- 7472559 TI - Role of N-methyl-D-aspartate receptor in acute spinal cord injury. AB - To clarify the role of N-methyl-D-aspartate (NMDA) receptors in acute spinal cord injury, changes in the intraspinal microcirculation after acute spinal cord injury in rabbits were examined. Systemic administration of MK-801, an NMDA receptor antagonist, at a dose of 5 mg/kg, significantly improved motor recovery after injury and significantly reduced edema formation at the injured site without altering spinal cord blood flow or vascular permeability at the injured site. These findings indicate that excitatory amino acids contribute to secondary spinal cord damage, especially edema formation, mediated by NMDA receptors in the early stage after injury. PMID- 7472560 TI - Electrocardiographic markers of abnormal left ventricular wall motion in acute subarachnoid hemorrhage. AB - A reversible and presumably neurogenic form of myocardial dysfunction may occur following subarachnoid hemorrhage (SAH), but the relationship of this finding to electrocardiographic abnormalities remains unclear. To clarify this issue, serial electrocardiograms (ECGs, mean 6.2 per patient) and echocardiograms (mean 3.4 days after SAH) were obtained in 57 SAH patients without preexisting cardiac disease. The goal was to determine which specific electrocardiographic changes, if any, reflect abnormal left ventricular wall motion in acute SAH. Wall motion abnormalities were identified in five (8%) of 57 patients. Four of these affected patients experienced hypotension (systolic blood pressure < 100 mm Hg) and three exhibited pulmonary edema within 6 hours of SAH, compared to none of the 52 patients with normal wall motion (p < 0.0001). Patients with abnormal wall motion were more likely than patients with normal echocardiograms to have symmetrical T wave inversion (five of five vs. seven of 52, p < 0.001) and severe (> or = 500 msec) QTc segment prolongation (five of five vs. three of 52, p < 0.001) on serial ECGs. These associations maintained their significance with analysis limited to single ECGs performed on or near the day of echocardiography. Abnormal wall motion was also associated with borderline (2% to 5%) creatine kinase MB elevation (five of five vs. three of 52, p < 0.001) and poor neurological grade (p < 0.0001). Although no combination of findings on a single ECG resulted in 100% sensitivity for abnormal wall motion, the presence of either inverted T waves or severe QTc segment prolongation on serial ECGs was associated with 100% sensitivity and 81% specificity. These results demonstrate an association between reduced left ventricular systolic function, mild creatine kinase MB elevation, and electrocardiographic repolarization abnormalities in acute SAH. Symmetrical T wave inversion and severe QTc segment prolongation best identified patients at risk for myocardial dysfunction and may serve as useful criteria for echocardiographic screening following SAH. PMID- 7472561 TI - A synthetic dural prosthesis constructed from hydroxyethylmethacrylate hydrogels. AB - Hydroxyethylmethacrylate (HEMA) hydrogels were investigated for their suitability as a dural prosthesis. Poly-HEMA has many characteristics required for an artificial dural substitute: it is durable, flexible, easily prepared, inexpensive, easily sterilized and handled, easily shaped, and known to be chemically inert and nontoxic. Sheets made of plain HEMA were evaluated as dural substitutes in rats and rabbits after either craniotomy or laminectomy with durectomy. Histological evaluations of the prostheses and the underlying tissues were undertaken at various time points from 2 to 9 weeks postoperatively. There was minimal tissue response to the implanted HEMA gel in contrast to marked thickening of the overlying leptomeninges and cortical herniation in the control animals. It is concluded that HEMA gels fulfill the essential criteria for an effective dural substitute. PMID- 7472562 TI - The microsurgical anatomy of the jugular foramen. AB - The microsurgical anatomy of the jugular foramen was studied in 10 fixed cadavers, each cadaver consisting of the whole head and neck. Five of the cadavers were injected with latex. The jugular foraminal region was exposed using the infratemporal fossa type A approach of Fisch and Pillsbury in five cadavers (10 sides) and the combined cervical dissection-mastoidectomy-suboccipital craniectomy approach in five cadavers (10 sides). The right foramen was larger than the left in seven cases (70%), equal in two cases (20%), and smaller in one case (10%). The dura covering the intracranial portal of the foramen had two perforations, a smaller anteromedial perforation through which passed the ninth cranial nerve (CN IX), and a larger posterolateral perforation, through which passed the 10th and 11th cranial nerves (CNs X and XI) and the distal sigmoid sinus. The perforations were separated by a fibrous septum in 16 specimens (80%). After exiting the posterior fossa, CNs IX, X, and XI all lay anteromedial to the superior jugular bulb (SJB) within the jugular foramen. The inferior petrosal sinus (IPS) entered the foramen between CNs IX and X in most cases; however, in 10% of our cases it entered the foramen between CNs X and XI, and in 10% it entered the foramen caudal to CN XI. The IPS terminated in the SJB in 90% of our cases; in 40%, the IPS termination consisted of multiple channels draining into both the SJB and internal jugular vein. This study shows that the arrangement of the neurovascular structures within the jugular foramen does not conform to the hitherto widely accepted notion of discrete compartmentalization into an anteromedial pars nervosa containing CN IX and the IPS and a posterolateral pars venosa containing the SJB, CNs X and XI, and the posterior meningeal artery. PMID- 7472563 TI - Early discovery of a traumatic carotid-cavernous sinus fistula by jugular venous oxygen saturation monitoring. Case report. AB - This report describes the early diagnosis, due to an acute increase of jugular venous oxygen saturation occurring 20 hours after trauma, of a traumatic carotid cavernous sinus fistula after severe head injury. Hyperemia in severe head injury should be treated only after an intracerebral arteriovenous communication has been excluded. PMID- 7472564 TI - Primary intraosseous meningioma. Case report. AB - Primary intraosseous meningioma of the skull is an uncommon lesion often confused preoperatively with a primary bone tumor of the skull. The case of an intraosseous meningioma without dural connection or association with a cranial suture is reported that was radiographically diagnosed as Paget's disease and initially treated conservatively. Persistent headache prompted a biopsy, yielding a benign, heavily ossified, and psammomatous meningioma. The lesion was treated with wide surgical excision and acrylic cranioplasty. Subsequent review of the literature has revealed 35 additional cases of purely intraosseous meningioma. An analysis of these 36 cases demonstrates a mean patient age at diagnosis of 45 years and a 2:1 female preponderance, with the majority of lesions associated with cranial sutures. Sixty-four percent of the lesions were hyperostotic on plain skull x-ray films; the rest were osteolytic or a mixture of both. There was no relationship to prior trauma. The treatment of choice is wide surgical resection followed by cranial reconstruction. PMID- 7472565 TI - Accessory middle cerebral artery. Case report. AB - An anomalous vessel is reported in a patient with an anterior circulation aneurysm. The presence of an accessory middle cerebral artery arising from the postcommunicating segment of the anterior cerebral artery is described. The clinical relevance and possible etiology of the anomaly are reviewed. PMID- 7472566 TI - Intracranial facial nerve rhabdomyoma. Case report. AB - Nerve rhabdomyomas are exceedingly rare benign tumors of the peripheral nerves consisting of well-differentiated striated muscle fibers admixed with parental nerve fibers. Only one case of intracranial nerve rhabdomyoma has been described, which affected the trigeminal nerve. This report presents the detailed neuropathological description of a nerve rhabdomyoma arising in the schwannian portion of the facial nerve root in a 41-year-old Caucasian man. The nerve fibers were arranged chaotically as in a traumatic neuroma. Because of the intimate intermingling of this slow-growing tumor with the parental nerve fibers, complete excision should be avoided. PMID- 7472567 TI - Magnetic resonance imaging of a glioblastoma of the optic chiasm. Case report. AB - Malignant optic glioma causing blindness was difficult to diagnose prior to the introduction of computerized tomography (CT) and magnetic resonance (MR) imaging, because earlier neuroradiological procedures often gave negative results and the clinical symptoms for this entity are not specific. In such cases only a craniotomy or postmortem examination revealed the tumor. The authors found no precise description in the literature of a malignant optic glioma diagnosed with modern imaging methods. They present a patient in whom biopsy results confirmed the CT and MR findings of glioblastoma multiforme of the optic chiasm. PMID- 7472568 TI - Mandibular subluxation as an adjunct to exposure of the distal internal carotid artery in endarterectomy surgery. Technical note. AB - The location of the carotid artery bifurcation and the distance atherosclerotic disease extends distally in the internal carotid artery (ICA) are two factors that contribute to the technical difficulty of carotid endarterectomy. When the bifurcation is high (above C-3) or the disease extends distally, standard approaches may not provide adequate exposure for dissection of plaque or for arteriotomy repair. A simple method of mandibular subluxation is described for added exposure of the distal carotid artery. The criteria for use of this method include: a carotid bifurcation at or above C-2; disease extending to within 2 cm of the skull base; and a small-caliber distal ICA lumen with the expectation of a patch graft extending close to the skull base. In dentulous patients, the mandible is subluxed by attaching an intradental wire from the ipsilateral mandibular bicuspid to an intradental wire around the contralateral maxillary bicuspid. In edentulous patients, a wire is placed around the ipsilateral mandible and secured to a wire placed through the anterior nasal spine. The entire preoperative subluxation requires 10 to 15 minutes under anesthesia and an additional 1 to 2 minutes postoperatively to remove the wires. A single skin suture and an absorbable intraoral suture were placed in some edentulous patients. This technique has been evaluated over a 15-month reference period during which 115 carotid endarterectomies were performed. The criteria stated above were met in seven cases (six patients, 6%) and jaw subluxation was performed preoperatively. An additional 1 to 2 cm of distal exposure was obtained by using this technique and endarterectomy proceeded without complication. A slight "shift" of the standard anatomical landmarks occurred due to the movement of the mandible, which was easily recognized. There were no significant postoperative complaints related to the subluxation; specifically, no temporomandibular joint pain, no other postoperative pain, and no tooth damage were encountered. It is concluded that this relatively simple approach to mandibular subluxation provided significant added exposure to the distal ICA without notably increasing operative time. In addition, there was no morbidity and little additional care was needed when compared with other more radical approaches to high carotid artery exposure. PMID- 7472569 TI - Galen's anecdote of the fallen sophist: on the certainty of science through anatomy. AB - In this paper the author recounts an anecdote presented by Galen of Pergamum (circa 130-200 A.D.) about a sophist named Pausanias, who fell from his mount and struck his back against a rock. The patient developed a subsequent loss of sensation in the fingers of his left hand with complete sparing of motor function. Numerous medications were applied to his hand but to no avail. Galen stated that he applied the same medications to the original point of dorsal tenderness, resulting in the patient's dramatic and full recovery. Galen attributed the healing to local drug action at the site of a presumed spinal root injury, at the level of C-7. Galen repeated this anecdote elsewhere to illustrate the remote effects of spinal cord and nerve injury and the importance of treating the site of pathology, rather than its somatic manifestations. Galen's observation is interpreted in light of his earlier experiments on spinal cord and nerve transections in live animals and his evolving concepts of functional and correlative neuroanatomy. This anecdote is also discussed as a striking example of the dangers of conjecture and the temptation to confuse association with causation when interpreting the effects of therapy in light of widely accepted paradigms. PMID- 7472570 TI - Luigi Rolando and his pioneering efforts to relate structure to function in the nervous system. AB - The fissure separating the motor from the sensory cortex and the substantia gelatinosa capping the posterior horn of the spinal cord are still known by the name of the Italian anatomist Rolando, Luigi Rolando was born in Turin, Italy, in 1773 and died in 1831. His life was not easy, the first of his problems being the death of his father when Rolando was still very young. Three people were to be influential in his life and career: Father Maffei, his maternal uncle who raised him; Dr. Cigna, the anatomy professor who discovered his talent; and Dr. Anformi, a general practitioner who introduced him to the practice of medicine and to the best circles of the city. Forced to leave Turin by the Napoleonic invasion of the country, Rolando first stopped in Florence, where he learned about anatomical dissection, drawing, and engraving and studied the appearance of nervous tissue under the microscope. Later he went to Sardinia where, although cut off from European cultural circles, he developed his major theories. Rolando pioneered the idea that brain functions could be differentiated and located in specific areas and discovered the fixed pattern of cerebral convolutions, highlighting motor and sensory gyri. He demonstrated the complexity of the central gray matter of the spinal cord, describing the "substantia gelatinosa," and he deduced that nervous structures are connected in a network of nervous fibers linked by electrical impulses. Rolando had to struggle for recognition, however, as the priority of his discoveries was challenged by the almost contemporaneous work of Gall and Spurzheim on cerebral localization and of Flourens on cerebellar function. Nevertheless, his efforts contributed greatly to the clarification of brain function. His observations on nervous anatomy have been especially accurate, as shown by the nomenclature "fissure of Rolando." PMID- 7472571 TI - Lateral discs and nerve sheath tumors. PMID- 7472572 TI - CART for prediction of function after head trauma. PMID- 7472573 TI - PEG-SOD after head injury. PMID- 7472574 TI - Cervical spondylotic myelopathy. PMID- 7472575 TI - Ischemic basis for essential hypertension? PMID- 7472576 TI - The future supply of molybdenum-99. PMID- 7472577 TI - Special issue. Cardiovascular nuclear medicine 1995, II. PMID- 7472578 TI - Utility of iodine-123-BMIPP in the diagnosis and follow-up of vasospastic angina. AB - Myocardial damage caused by vasospastic angina (VSA) may be detected by [123I]BMIPP, a beta-methyl-branched fatty acid. We investigated whether BMIPP could be used in the diagnosis and follow-up of patients with VSA. METHODS: Thirty-two patients with VSA were studied with resting BMIPP-SPECT in comparison to stress perfusion imaging with either 201Tl or 99mTc-MIBI. During coronary arteriography, spasm was induced by provocative testing with acetylcholine or ergonovine, and only total or subtotal occlusion was considered positive. Decreased BMIPP uptake was semiquantitatively evaluated segmentally aided by polar map display. RESULTS: Reduced BMIPP uptake was observed in 25 of 32 patients (78%), with complete or partial agreement between the BMIPP abnormality and coronary territory seen in 23 patients (72%). In contrast, a perfusion abnormality was seen in only 10 patients (31%). In the repeat BMIPP study (n = 23) during the follow-up period (average 206 days), 11 of 14 patients who showed BMIPP improvement also had improved angina attacks. Conversely, two of nine patients with nonimproved BMIPP showed improved symptoms (p < 0.05). CONCLUSION: BMIPP can detect myocardial injury associated with VSA and may be useful in monitoring response to treatment. PMID- 7472579 TI - Motion correction in exercise first-pass radionuclide ventriculography without an external point source. AB - Exercise first-pass radionuclide ventriculography provides valuable diagnostic and prognostic information in patients with coronary artery disease. In this procedure, motion correction of the images is commonly performed using a second external point source attached to the chest wall during exercise (dual-isotope method). Recently, a motion correction algorithm without an external point source (single-isotope method) was developed and the results compared with those of the dual-isotope method. METHODS: To examine the accuracy of the motion correction method, a phantom study was performed using a moving cardiac phantom with a motion speed of up to 169 cycle/min and motion amplitude up to 6 cm. Count fluctuation in the phantom region by motion was calculated as a coefficient of variation (CV). In the clinical study, time-activity curves of the left ventricular phase were created for quantitative assessment of variation as CV values of the ejection fraction in the central five cardiac cycles after correction by the two methods during exercise radionuclide ventriculography in 17 patients. RESULTS: In the moving phantom, both the single- and dual-isotope methods reduced the CV values less than 10%. In the clinical study, the single isotope method provided less CV value of ejection fraction (9.8% +/- 5.6%) than the dual-isotope method (24.8% +/- 10.5%) (p < 0.01), indicating less individual variation of ejection fraction values. CONCLUSION: These data indicate that object motion can be accurately corrected in the moving phantom by both single- and dual-isotope methods. In clinical studies, the single-isotope method is more accurate. PMID- 7472580 TI - Enhanced detection of viable myocardium by technetium-99m-MIBI imaging after nitrate administration in chronic coronary artery disease. AB - The aim of this study was to assess whether nitrate administration improves the imaging capabilities of 99mTc-MIBI tomography in detecting viable myocardium in coronary artery disease (CAD). METHODS: Thirty-one patients with angiographically proven CAD and chronic LV dysfunction (ejection fraction 39% +/- 9%) underwent two 99mTc-MIBI studies on separate days: one under rest conditions and the other after nitroglycerine (0.005 mg/kg per os) administration. Within 1 wk, all patients also underwent rest-redistribution 201Tl imaging. Eight patients were also studied by echocardiography before and 5 +/- 3 mo after coronary revascularization. RESULTS: On resting 99mTc-MIBI images, 302 segments had normal uptake, 183 segments had moderately reduced uptake and 197 had severely reduced uptake. Of the segments with severely reduced uptake, 54 (27%) had increased uptake after nitroglycerine and were viable on 201Tl images. Of the 143 (73%) segments with severely reduced 99mTc-MIBI uptake and no change after nitroglycerine, 81% were nonviable on 201Tl images. In the eight patients studied before and after revascularization, 87% of segments with reversible 99mTc-MIBI defects and abnormal LV function demonstrated functional recovery after revascularization, whereas 89% of segments with irreversible 99mTc-MIBI defects did not. CONCLUSION: In patients with chronic ischemic LV dysfunction, nitrate administration improved the detection of severely hypoperfused but still viable myocardium on 99mTc-MIBI images. PMID- 7472581 TI - Myocardial viability index in chronic coronary artery disease: technetium-99m methoxy isobutyl isonitrile redistribution. AB - The purpose of this study was to evaluate whether an additional redistribution image after a rest 99mTc-MIBI injection enhances detection of viable myocardium and predicts functional recovery after coronary revascularization in patients with chronic coronary artery disease (CAD). METHODS: Thirty-one patients (29 men, mean age 55 +/- 10 yr) with proven CAD and left ventricular (LV) dysfunction (ejection fraction 39% +/- 9%) underwent resting 99mTc-MIBI tomography with initial (1 hr) and delayed (5 hr) images. Within 1 wk of MIBI imaging, all patients underwent rest-distribution 201Tl imaging. Eight patients also underwent two-dimensional echocardiography before and 5 +/- 3 mo after coronary revascularization. RESULTS: On the initial 99mTc-MIBI images, 302 myocardial segments were normal, 183 showed moderate and 197 severe reduction of tracer uptake. Of these 197 segments, 47 (24%) demonstrated increased tracer uptake (> or = 10% versus initial) on delayed images (from 43% +/- 8% to 60% +/- 8%, p < 0.001) and were considered as showing 99mTc-MIBI redistribution. These 47 segments were observed in 20 (65%) patients in whom 201Tl images detected viable myocardium in the same segments. In the eight patients studied before and after revascularization, 83% of segments with 99mTc-MIBI redistribution and abnormal LV function showed functional recovery after revascularization, while 96% of segments without 99mTc-MIBI redistribution did not show functional recovery. CONCLUSION: Resting 99mTc-MIBI redistribution frequently occurs in patients with chronic CAD. Acquisition of 99mTc-MIBI redistribution images enhances detection of viable myocardium and predicts functional recovery after revascularization. PMID- 7472582 TI - Myocardial viability assessment with technetium-99m-tetrofosmin and thallium-201 reinjection in coronary artery disease. AB - Exercise-rest 99mTc-tetrofosmin myocardial perfusion images with a 2-day protocol was compared to exercise-redistribution-reinjection 201Tl images to assess the ability of 99mTc-tetrofosmin to detect viable myocardium. METHODS: We studied 25 patients with coronary artery disease and regional or global left ventricular dysfunction. Myocardial SPECT images with 99mTc-tetrofosmin were obtained 10 min after injection during exercise and 1 and 3 hr after rest injection. Within 1 wk of the 99mTc-tetrofosmin study, exercise-redistribution-reinjection 201Tl SPECT imaging was performed. RESULTS: Visual analysis demonstrated concordance between 201Tl and 99mTc-tetrofosmin imaging for defect reversibility in 126 of 209 segments (60%), with initial defects on both exercise 201Tl and 99mTc-tetrofosmin images. In the remaining discordant 83 segments (40%), 73 (88%) appeared nonreversible on 99mTc-tetrofosmin imaging but were reversible on 201Tl imaging. CONCLUSION: On the basis of defect reversibility by visual analysis, exercise rest 99mTc-tetrofosmin imaging underestimates myocardial viability compared to 201Tl reinjection imaging. The identification of viable myocardium with both 99mTc-tetrofosmin and 201Tl can be greatly enhanced to a similar degree if the severity of reduction in activity within nonreversible defects is considered. These two agents may provide comparable information about myocardial viability by quantitative analysis of defect severity. PMID- 7472583 TI - Reverse redistribution in resting thallium-201 myocardial scintigraphy in chronic coronary artery disease: an index of myocardial viability. AB - The aim of this study was to evaluate whether segments with reverse redistribution on rest-redistribution 201Tl scintigraphy represent viable tissue or scar. METHODS: Nineteen patients (17 men, 2 women; mean age 53 +/- 8 yr) with coronary artery disease underwent rest-redistribution 201Tl study before coronary revascularization. Regional 201Tl uptake was analyzed quantitatively. Regional left ventricular wall motion was assessed before and after coronary revascularization using two-dimensional echocardiography and a three-point scale (1 = normal, 2 = hypokinetic, 3 = akinetic/dyskinetic). Two patterns of reverse redistribution were identified: pattern with normal 201Tl uptake in rest and abnormal in redistribution images and pattern with abnormal 201Tl uptake in rest and a significant decrease in redistribution images. RESULTS: Of the 247 segments analyzed, 85 were classified as normal, 37 as reversible defects, 83 as fixed defects and 42 as reverse redistribution (19 RR-A, 23 RR-B). Segments with RR-A differed from those with RR-B in wall motion score (1.4 +/- 0.7 versus 2.0 +/- 1.0). Electrocardiographic Q-waves were present in 26% of segments with RR-A and in 57% of segments with pattern B. After revascularization, all dyssynergic segments with pattern A showed improved wall motion, while only 40% of segments with pattern B and abnormal wall motion had such improvement. CONCLUSION: Our results suggest that dyssynergic segments with pattern A should be considered viable, while more caution should be used in classifying those with pattern B. PMID- 7472584 TI - Decreased uptake of iodinated branched fatty acid analog indicates metabolic alterations in ischemic myocardium. AB - We previously reported that uptake of 123I-labeled 15-iodophenyl 3-methyl pentadecanoic acid (BMIPP) was lower than that of thallium in ischemic myocardium. Such discordant findings between BMIPP and thallium were compared with those of PET using 18F-deoxyglucose (FDG) and 11C-acetate to assess metabolic alterations in such segment. METHODS: Sixteen patients with coronary artery disease underwent both BMIPP SPECT and PET. Relative FDG uptake (% FDG uptake) and the clearance rate constant (% Kmono) of 11C-acetate from the myocardium were calculated as markers of glucose and oxidative metabolism, respectively. RESULTS: Relative FDG uptake of the myocardial segments with reduced BMIPP uptake and normal thallium uptake (discordant segments) was similar (85.3 +/- 10.3) to that of the normal segments (86.5 +/- 11.7) but higher than that of segments with reduced uptake of both BMIPP and thallium (67.5 +/- 19.9). Similarly, the discordant segments showed a higher % Kmono value (77.8 +/- 13.1 versus 70.0 +/- 19.1) and FDG-to-perfusion ratio (1.15 +/- 0.08 versus 1.01 +/- 0.22) than in the concordantly reduced segments. CONCLUSION: BMIPP uptake appears to provide metabolic information independent of thallium uptake. Combined imaging of BMIPP and thallium may potentially identify ischemic but viable myocardium. PMID- 7472585 TI - Myocardial oxidative metabolism in hyperthyroid patients assessed by PET with carbon-11-acetate. AB - Hyperthyroid patients often complain of cardiovascular symptoms because of increased metabolism. This study was designed to quantitatively evaluate myocardial oxidative metabolism in these patients. METHODS: Dynamic PET with 11C acetate was performed in 19 patients who had not undergone treatment for hyperthyroidism. Eight were restudied 2 wk after oral administration of propranolol. The clearance rate constant of 11C-acetate (Kmono) was calculated with monoexponential fitting of tracer washout from the myocardium as a marker of myocardial oxidative metabolism. The results were compared with those in nine normal subjects both at rest and during dobutamine infusion. RESULTS: Kmono in our patients (0.109 +/- 0.028 min-1) was significantly increased compared to normal subjects (0.066 +/- 0.016 min-1) (p < 0.05). After propranolol treatment, Kmono decreased (0.082 +/- 0.014 min-1) but remained significantly higher in eight patients than normal subject levels (p < 0.05), while the rate pressure product decreased significantly (7500 +/- 1700) toward the normal range (7900 +/- 1500). CONCLUSION: These results suggest the possibility of excessive myocardial oxygen consumption in hyperthyroid patients. The clearance rate of 11C-acetate is a new and valuable index to assess myocardial oxidative metabolism not closely related to the pressure rate product or thyroid hormones in these patients. PMID- 7472586 TI - Prediction of improvement in left ventricular function with iodine-123-IPPA after coronary revascularization. AB - Iodine-123-phenylpentadecanoic acid (IPPA) is a synthetic fatty acid suitable for myocardial imaging. This study is the result of a Phase I/II trial to evaluate IPPA's ability to predict functional recovery in patients undergoing coronary revascularization. METHODS: Twenty-three patients with documented coronary disease underwent sequential SPECT imaging with IPPA before and radionuclide ventriculography both before and 8 wk after revascularization. Software was developed to evaluate myocardial IPPA metabolism and to determine the fraction of the left ventricle with intermediate metabolism. RESULTS: There was a significant correlation between initial IPPA uptake and final LVEF. The fractional area of the left ventricle demonstrating IPPA metabolism in the intermediate metabolic range was significantly higher in patients who demonstrated a 5% or greater increase in EF after revascularization (0.90 +/- 0.08 versus 0.78 +/- 0.17, p = 0.04). When only the patients who received complete revascularization were evaluated, there was a more significant difference (improved 0.92 +/- 0.05 versus 0.74 +/- 0.17, p = 0.011). Taking a lower limit of 1 s.d. from the mean, (87%) the six patients who had > or = 5% increase in LVEF after revascularization had more than 87% of the left ventricle in the intermediate metabolic range, whereas seven of ten patients whose change in LVEF was < 5% had less than 87% in the intermediate metabolic range (p = 0.011). CONCLUSION: In this initial experience, the amount of myocardium in the intermediate metabolic range is associated with improvement in LVEF after revascularization, especially in patients receiving complete revascularization. PMID- 7472587 TI - Technetium-99m-sestamibi imaging with nitrate infusion to detect viable hibernating myocardium and predict postrevascularization recovery. AB - We tested the relationship of nitrate-induced changes in 99mTc-sestamibi perfusion tomography and first-pass radionuclide angiocardiography (FPRNA) with postrevascularization functional recovery of asynergic territories. METHODS: Twenty-eight patients, all with prior infarction and left ventricular dysfunction, underwent two 99mTc-sestamibi rest studies: one under baseline conditions and the other with nitrate infusion. The baseline study was repeated after revascularization. Changes in global and regional perfusion and ventricular function were evaluated by perfusion tomography and FPRNA. Hibermating myocardium was identified by functional recovery in postrevascularization FPRNA. RESULTS: Eleven patients and 31 segments in 19 coronary territories had functional recovery. Nitrate-induced FPRNA changes showed poor agreement with postrevascularization modifications: kappa = 0.24, ns, for the global and kappa = 0.32, p < 0.01 for regional function. The agreement between nitrate-induced and postrevascularization perfusion changes was good both considering the patients (k = 0.57, p < 0.01) and the abnormal coronary territories (k = 0.63, p < 0.0005). The agreement of nitrate-induced perfusion changes with postrevascularization functional outcome was excellent on a patient (k = 1, p < 0.0005), and coronary territory basis (k = 0.82, p < 0.0005). Conversely, the agreement between nitrate and postrevascularization FPRNA was poor: k = 0.18, ns. CONCLUSION: These data suggest that the nitrate-induced changes in 99mTc-sestamibi perfusion imaging are useful to detect hibernating myocardium which recovers its function after revascularization. PMID- 7472588 TI - Preventing patient motion during tomographic myocardial perfusion imaging. AB - We evaluated whether use of a device that positions and supports the upper extremities and back during tomographic myocardial perfusion imaging reduces the incidence and severity of patient motion and patient motion artifact. METHODS: We enrolled 190 patients referred for stress/redistribution myocardial perfusion imaging. All patients were imaged once with the patient support device (PSD) and once without it. Patients were randomly assigned to use the PSD either during poststress or redistribution imaging. The presence and severity of patient motion was determined by visual inspection and quantitative motion detection. The presence of reconstruction artifact due to motion was detected visually and confirmed by motion correction. RESULTS: Use of the PSD reduced the incidence of motion from 38% +/- 3.7% to 26% +/- 3.3% (p < 0.05) and reduced the amount of motion by 5.3 +/- 2.2 mm (p < 0.05). Patients who did move, moved less when using the PSD by both visual (p < 0.02) and quantitative criteria (p < 0.05). Use of the PSD reduced the incidence of reconstruction artifacts to one third of control (p < 0.05). CONCLUSION: The use of this positioning and support device during tomographic myocardial perfusion imaging reduces the incidence and severity of patient motion and motion artifact. PMID- 7472589 TI - Pulmonary perfusion after endovascular stenting of pulmonary artery stenosis. AB - Pulmonary artery stenosis is a well-known condition after surgical correction of tetralogy of Fallot. Endovascular stenting of the stenosis is a new technique for correction without surgical intervention. Objective evaluation of the procedure, however, is often hampered by moderate or severe pulmonary valve insufficiency. This disadvantage does not apply to 99mTc-macroaggregates of albumin (MAA) scintigraphy of the lungs. Moreover, quantification can be performed relatively easy. METHODS: Seven patients with surgically corrected tetralogy of Fallot (4 men, 3 women, mean age 15.7 yr, range 5-24 yr) were studied. The mean diameter decrement of a pulmonary artery was 69% +/- 8.7%. Before and after stenting, relative uptake in the left and right lung was assessed after injection of 37-55 MBq 99mTc-MAA. Three patients were studied twice after stenting. RESULTS: In all patients, perfusion of the affected lung increased significantly: before 22.7% +/ 10.8%, after 38.6% +/- 12.3% (p < 0.0001). All patients claimed clinical improvement of their condition after stenting. The perfusion gain did not correlate with the prestenting diameter decrement or with the pressure gradient over the stenosis. Lung uptake in the patients studied twice after stenting was similar between the initial and poststenting study (< or = 2% difference). CONCLUSION: Quantitative 99mTc-MAA lung imaging is a noninvasive technique without patient discomfort that objectively measures the effect of stenting pulmonary artery stenosis. The final outcome of stenting with regard to increment of pulmonary blood flow is not predicted by the severity of the stenosis or the pressure gradient over the stenosis. PMID- 7472590 TI - Technetium-99m-tetrofosmin in dipyridamole-stress myocardial SPECT imaging: intraindividual comparison with technetium-99m-sestamibi. AB - Tetrofosmin is a new 99mTc-labeled myocardial perfusion imaging agent. Biodistribution studies suggest more favorable heart-to-adjacent organ biokinetics than for 99mTc-sestamibi after injection during exercise. The aim of this work was to determine intraindividually whether tetrofosmin is more suitable than sestamibi for pharmacological stress testing in a 1-day protocol. METHODS: Thirty subjects underwent two similar 1-day, rest and dipyridamole stress imaging protocols: one using tetrofosmin, the other using sestamibi. SPECT was performed 60 min after tracer administration. Myocardial images were analyzed both visually and quantitatively. RESULTS: Heart-to-liver activity ratios measured on the anterior SPECT projections were significantly higher for tetrofosmin than for sestamibi in the rest and stress studies. Heart-to-lung ratios were similar for both tracers. Significant linear correlations between tetrofosmin and sestamibi perfusion indices were found in normals and in patients with proven or suspected coronary artery disease. In segments showing abnormal uptake during stress, the perfusion indices were similar for tetrofosmin and sestamibi at rest and during stress. The degree of reversibility in these segments was also similar for both tracers. Finally, the extent, intensity and severity of perfusion defects were similar for both tracer studies. CONCLUSION: Tetrofosmin has a more optimal biodistribution than sestamibi when used in a 1-day, rest and dipyridamole stress myocardial SPECT imaging protocol. No significant difference in either the quality or diagnostic interpretation of the images could be demonstrated. PMID- 7472591 TI - Effect of caffeine on myocardial blood flow at rest and during pharmacological vasodilation. AB - Stress testing with intravenous injection of dipyridamole is frequently used for noninvasive detection of coronary artery disease (CAD) with PET or SPECT. Dietary intake of caffeinated food, beverages or medication might alter both resting and dipyridamole-induced hyperemic blood flow, thereby compromising the diagnostic sensitivity of dipyridamole stress testing. METHODS: To quantify the effect on myocardial blood flow at rest and during intravenous injection of dipyridamole, 12 healthy volunteers (mean age 27 +/- 6 yr) with low risk for CAD were studied with dynamic PET and a tracer kinetic model for 13N-ammonia after 24 hr of caffeine abstinence and after caffeine intake. RESULTS: Caffeine tended to increase the rate pressure product from 6873 +/- 1494 to 7566 +/- 1102 (p = 0.051), whereas resting myocardial blood flow remained unchanged (0.61 +/- 0.13 versus 0.58 +/- 0.07 ml/g/min, p = ns). The heart rate response to dipyridamole was inversely related to serum caffeine levels. Hyperemic blood flow (2.01 +/- 0.46 versus 1.31 +/- 0.0.38 ml/g/min; p < 0.001) and flow reserve (3.4 +/- 0.8 versus 2.3 +/- 0.7; p < 0.001) were inversely related to the caffeine dose. Coronary vascular resistance at rest tended to increase (132 +/- 32 versus 147 +/ 25 mmHg/ml/g/min; p = 0.06), whereas minimal coronary vascular resistance was significantly higher after caffeine (41 +/- 9 to 69 +/- 25 mmHg/ml/g/min; p < 0.01). CONCLUSION: Caffeine intake alters the coronary vasomotor tone at rest, which might lower the threshold for ischemic events in patients with CAD. It reduces hyperemic blood flow and flow reserve and the dipyridamole-induced increase in heart rate in a dose-dependent fashion. These findings emphasize the importance of carefully screening patients for intake of caffeinated food, beverages or medication prior to dipyridamole stress testing. PMID- 7472592 TI - Scintigraphic recording of blood volume shifts. AB - A scintigraphic device consisting of small scintillation probes tightly fixed to the skin was developed to record intravascular blood volume shifts continuously and simultaneously at several sites. The aim of the present study was to ascertain the reliability of the measurements obtained, show the blood shifts induced by common daily activities in healthy subjects and clarify the mechanisms responsible for hemodynamic changes. METHODS: Measurements were made in three fields of the right lung, the liver, thighs and calves of 16 men during Valsalva maneuver, hyperventilation, various posture changes and treadmill walking. Some tests were repeated. RESULTS: The measured blood volume shifts were always in the expected direction and in accordance with those reported in the literature; they also were reproducible. Therefore, the measurements were reliable. A pattern of blood volume changes peculiar to some common daily activities was recorded in subjects moving freely. Insights were obtained on the mechanisms responsible for the blood volume shifts. CONCLUSION: Our noninvasive technique provides reliable continuous measurements of blood volume changes at several sites during common daily activities and could be applied not only in healthy subjects but also in patients. PMID- 7472593 TI - Effect of aging on myocardial perfusion reserve. AB - Myocardial perfusion reserve (hyperemic divided by basal myocardial blood flow) describes vasodilator responsiveness of coronary-resistive vessels. The effect of aging and gender on myocardial perfusion reserve remains controversial. METHODS: We studied 56 normal volunteers (21 women, 35 men; aged 50 +/- 20 yr, range 21-86 yr) with 15O-water PET to measure myocardial blood flow during basal and hyperemic states with intravenous dipyridamole (0.56 mg/kg, n = 46) or adenosine (140 micrograms/kg/min, n = 10). For comparative analysis, patients were grouped according to age: < 30 yr (n = 11), 30-49 yr (n = 18), 50-69 yr (n = 15) and > or = 70 yr (n = 12). RESULTS: Overall, basal flow was 1.00 +/- 0.26 ml/min/g and hyperemic flow was 3.31 +/- 1.38 ml/min/g, resulting in a myocardial perfusion reserve of 3.38 +/- 1.35. There was an increase in basal flow with age (r = 0.45, p < 0.025), although hyperemic flow was only lower in patients > or = 70 yr, causing a significant reduction in myocardial perfusion reserve: 3.54 +/- 0.96 in < 30 yr, 4.23 +/- 1.35 in 30-49 yr, 3.51 +/- 1.21 in 50-69 yr and 1.94 +/- 0.46 in > or = 70 yr (p < 0.05 versus all groups < 70 yr). CONCLUSION: Myocardial blood flow during basal and hyperemia conditions are roughly comparable up to 60 yr of age. Above this age, there is significant increase in basal flow associated with an increase in systolic blood pressure. Above 70 yr, there is a significant reduction in hyperemic flow, and thus myocardial perfusion reserve independent of hemodynamic response to vasodilator stress. PMID- 7472594 TI - Enhanced cardiac clearance of iodine-123-MIBG in chronic renal failure. AB - We studied myocardial accumulation and clearance of [123I]metaiodobenzylguanidine (MIBG) along with measurements of plasma norepinephrine concentrations. METHODS: Myocardial imaging with MIBG and plasma norepinephrine concentration measurements were performed in 21 patients with chronic renal failure on dialysis and 11 control subjects. Dynamic acquisitions, begun immediately after MIBG injection, and planar images 15 and 150 min after injection were used to measure early and late myocardial uptake and clearance rates of MIBG from the heart. RESULTS: Early and late MIBG uptake was not significantly different from that in controls but MIBG clearance was significantly more rapid in controls. Based on echocardiographic findings, the patients were divided into three subgroups: those without left ventricular hypertrophy and dysfunction (n = 10), with hypertrophy (n = 4) and with dysfunction (n = 7). Among controls and the three patient subgroups, MIBG clearance was significantly higher in all the subgroups and also was significantly higher in patients with dysfunction than in those without hypertrophy and dysfunction. Plasma norepinephrine levels were significantly higher in patients than in controls and correlated significantly with the clearance rate. CONCLUSION: MIBG clearance from the heart was rapid in patients with chronic renal failure on dialysis, particularly those with left ventricular dysfunction or hypertrophy, suggesting cardiac sympathetic overactivity in these patients. PMID- 7472595 TI - Indium-111-antimyosin and iodine-123-MIBG studies in early assessment of doxorubicin cardiotoxicity. AB - Detection of myocyte cell damage with 111In-antimyosin and impairment of adrenergic neuron function with [123I]MIBG during doxorubicin administration may provide easy identification of patients at risk of significant functional impairment. METHODS: We studied 36 cancer patients who underwent chemotherapy, including doxorubicin, to assess [123I]MIBG and 111In-antimyosin uptake in the course of doxorubicin administration. MIBG scans, antimyosin scans and ejection fraction measurements were performed before chemotherapy, at intermediate cumulative doses and at maximal cumulative doses of doxorubicin. MIBG uptake was quantified by a heart-to-mediastinum ratio and antimyosin uptake was quantified by a heart-to-lung ratio. RESULTS: All patients had absent antimyosin uptake (mean ratio 1.40 +/- 0.06) with normal MIBG uptake (ratio 1.85 +/- 0.29) before chemotherapy; ejection fraction was 61% +/- 8%. With a 240-300 mg/m2 dose of doxorubicin, an increase in antimyosin uptake was observed with a ratio of 1.85 +/- 0.2 (p < 0.01), whereas a similar degree of MIBG uptake was observed (mean ratio of 1.80 +/- 0.2, p = ns); ejection fraction was 59% +/- 5% (p = ns). At 420 600 mg/m2, increased antimyosin uptake was observed with a ratio of 2.02 +/- 0.3 (p < 0.01), and a decrease in MIBG uptake was also observed (mean ratio of 1.76 +/- 0.2, p < 0.05); ejection fraction was 52% +/- 8% (p < 0.05). Patients with more intense antimyosin uptake at intermediate doses tended to be those with more severe functional impairment at maximal cumulative doses. CONCLUSION: At cumulative doses of 420-600 mg/m2, antimyosin and MIBG studies detect cell damage and impaired adrenergic neuron activity in patients with maintained or slightly decreased ejection fraction. PMID- 7472596 TI - Detection of perioperative coronary vasospasm on iodine-123-MIBG, thallium-201 and iodine-123-BMIPP myocardial SPECT images. AB - A 64-yr-old woman, who had no history of anginal attack, underwent cholecystectomy under general anesthesia. A few hours after successful surgery, the EKG showed T-wave inversions with QT-interval prolongations and torsades de pointes. The [123I]MIBG scintigraphic findings revealed marked denervation in the anteroseptal and inferoposterior myocardium, whereas 201TI myocardial SPECT showed only slightly reduced tracer uptake in those areas. The [123I]BMIPP scans showed abnormal fatty acid metabolism in the anteroseptal myocardium. Coronary angiogram detected no fixed stenosis, but coronary vasoconstriction in the left anterior descending artery was induced by intracoronary injection of acetylcholine. In our patient, the findings of cardiac imaging with [123I]MIBG, 201TI and [123I]BMIPP led to coronary angiography and the final diagnosis of probable perioperative coronary vasospasm. PMID- 7472597 TI - Reverse redistribution on dynamic exercise and dipyridamole stress technetium-99m MIBI myocardial SPECT. AB - Reverse redistribution on 201TI-chloride stress-redistribution myocardial scintigraphy has been associated with coronary artery stenosis. We report a patient whose two separate 99mTc-MIBI myocardial SPECT stress studies (dynamic exercise and dipyridamole) showed septal reverse redistribution and fixed inferior defect. Echocardiograms showed left ventricular (LV) hypertrophy and diffuse hypokinesis, especially in the inferior wall, and EKG showed LV hypertrophy and strain and inferior infarct. Coronary angiogram confirmed two vessel disease involving 80%-90% stenosis of the proximal second diagonal branch of the left anterior descending artery and 75%-90% stenosis of the right coronary artery as well as global left ventricular dysfunction. Reverse redistribution on 99mTc-MIBI myocardial SPECT occurring on dynamic or dipyridamole stress may indicate damaged but viable myocardium. PMID- 7472598 TI - Metabolic mismatch of septal beta-oxidation and glucose utilization in left bundle branch block assessed with PET. AB - Preserved septal uptake of the new long-chain fatty acid analog [18F]FTHA was found in a patient with three-vessel disease, a history of previous anterior myocardial infarction and left bundle branch block (LBBB), despite severely decreased septal [18F]FDG uptake that suggested scarred tissue. Nearly absent [19F]FDG uptake in the septum could not be explained by concordant reduction of septal perfusion as assessed by 99mTc-MIBI SPECT. These data may point to divergent metabolic effects of the conduction abnormality in LBBB with consecutively reduced septal exogenous glucose utilization but unaffected septal beta-oxidation. PMID- 7472600 TI - Effect of monounsaturation of a branched fatty acid on organ selectivity. AB - 1-[11C]-3-R,S-methylheptadecanoic acid (BMHA) is a branched chain fatty acid analog that is transported into the myocardium. Due to incomplete metabolism, however, radiolabeled products are trapped within myocytes. Recently, we demonstrated that this compound is an excellent tracer to monitor fatty acid metabolism. METHOD: To evaluate the effect of mono-unsaturation on myocardial substrate utilization, we prepared 1-[11C]-3-R,S-methyl-trans--heptadec-7-enoic acid (t-7-BMHA) and measured its biodistribution in rats. In addition, preliminary PET studies were performed on dogs. RESULTS: Biodistribution studies demonstrated that myocardial-to-lung and myocardial-to-blood ratios for t-7-BMHA are higher than those for BMHA. Fifteen minutes after injection, heart-to-lung ratios were 5.23 compared to 2.92 and heart-to-liver ratios were 3.07 compared to 1.41 for t-7-BMHA and BMHA. By 30 min postinjection heart-to-lung ratios were 7.03 compared to 5.88 and heart-to-liver ratios were 4.43 compared to 1.09. The heart-to-blood ratio of t-7-BMHA was greater than 11:1. PET imaging with 1-[11C] t-7-BMHA demonstrated high myocardial extraction, prolonged retention of radioactivity and excellent image quality. Accumulation of radioactivity in the myocardium reached a plateau within 10 min postinjection, with heart-to-blood ratios exceeding 20:1 and heart-to-lung ratios exceeding 10:1. Blood clearance of radioactivity was biphasic with half-times of 1.46 and 14.7 min, respectively. CONCLUSION: These data suggest that introduction of a trans-double bond in BMHA improves myocardial selectivity and results in a potentially superior imaging agent. PMID- 7472599 TI - Atrophic nerve fibers in regions of reduced MIBG uptake in doxorubicin cardiomyopathy. AB - A myocardial MIBG-SPECT examination was conducted 2 wk after doxorubicin chemotherapy on a 52-yr-old woman without cardiac symptoms. Despite normal 201TI scintigraphy, reduced MIBG uptake was detected in the apical anterior, inferior and lateral segments of the left ventricle. The patient died of congestive heart failure due to doxorubicin-induced cardiomyopathy 10 mo later. At necropsy, the left ventricle was markedly dilated and the apical anterior, inferior and lateral walls were thin, stiff and whitish. Nerve fibers in the apical inferior wall were atrophic and markedly fibrotic where MIBG uptake was most reduced. Nerve fibers in the septum were normal where MIBG uptake had remained normal. The histologic findings correspond with the findings on the MIBG image. MIBG imaging may detect cardiac sympathetic denervation in doxorubicin-induced cardiomyopathy before cardiac symptoms are manifest and cardiac function deteriorates. PMID- 7472601 TI - Assessment of adrenergic neuron function altered with progression of heart failure. AB - We used MIBG to evaluate cardiac adrenergic neuron integrity and function in congestive heart failure. METHODS: Rats were treated with adriamycin (2 mg/kg, s.c.) once a week for 7, 8 and 9 wk. In analyzing cardiac adrenergic neuron function, we assessed alterations of uptake-1, exocytotic release and nonexocytotic metabolic release in relation to progression of heart failure. RESULTS: LVEF progressively decreased. Cardiac MIBG accumulation (4 hr postinjection) decreased to 53% of control at 7 wk and markedly decreased to 14% of control at 9 wk, accompanied by massive pleural effusions. Reduction of MIBG accumulation in the lung and spleen, which are adrenergic-rich organs similar to the heart, were less pronounced compared to reduction in the heart. There was no difference in cardiac uptake of 3H-norepinephrine between the control and 8-wk groups. Cardiac uptake of 3H-norepinephrine decreased 91.0% in the control and 90.8% in the 8 wk group by pretreatment of desipramine, indicating no difference in the uptake-1 component. CONCLUSION: Congestive heart failure due to adriamycin cardiomyopathy progressively accelerates exocytotic release of norepinephrine predominantly from cardiac adrenergic neurons, but neuronal uptake function is not disturbed so long as heart failure is not advanced. In the advanced stage, nonexocytotic metabolic release is induced specifically in cardiac adrenergic neurons due to energy depletion and norepinephrine release markedly increases. PMID- 7472602 TI - Subcellular distribution of technetium-99m-N-NOEt in rat myocardium. AB - The aim of this study was to determine the subcellular distribution of bis(N ethoxy N-ethyl)dithiocarbamato nitrido technetium(V) (99mTcN-NOEt) in rat heart by differential centrifugation techniques. Extraction of the activity from homogenized rat heart tissue was also performed to assess whether myocardial retention might induce changes in the chemical identity of the complex. METHODS: Anesthetized rats were intravenously injected with 99mTcN-NOEt, the heart tissue was extracted and homogenized and tissue fractions were obtained by differential centrifugation. The efficiency of organelle separation was determined by assay of each centrifugal fraction using enzyme markers. Lactate dehydrogenase (LDH), acid phosphatase (ACP), alkaline phosphatase (ALP) and 5'-nucleotidase (5'ND) activities were assayed using standard spectrophotometric methods. Succinic dehydrogenase (SDH) activity was determined using a p-iodo-nitrotetrazolium linked assay. Severe cell membrane and organelle disruption were induced by prolonging the homogenization time and their effect on the subcellular distribution of 99mTcN-NOEt was studied. The activity from homogenized heart tissue was extracted using the Folch technique and analyzed by TLC and HPLC. RESULTS: Most of the 99mTcN-NOEt activity was found to be associated with the hydrophobic components of the cell. No evidence of specific association of activity with the cytosolic and mitochondrial components was observed. Organelle and membrane cleavage did not cause release of activity into the cytosol. Approximately 90% of 99mTcN-NOEt activity was extracted from ventricular tissue and the chemical nature of 99mTcN-NOEt was not altered by uptake by myocardium. CONCLUSION: Cell membranes are the most apparent site of localization of 99mTcN NOEt in heart tissue. PMID- 7472603 TI - Effects of scatter correction on the measurement of infarct size from SPECT cardiac phantom studies. AB - Thallium-201 and 99mTc-sestamibi images of the heart contain a significant amount of scattered events which degrade image quality. Newer generation gamma cameras exhibit enhanced energy resolution and hardware/software to perform scatter correction. The principal aim of this study was to evaluate the effects of these advances in instrumentation on the quantitation of defect size from tomographic images of the heart obtained from a cardiac phantom. METHODS: Tomographic images of a cardiac phantom containing no defect and defects of 5%-70% of total myocardial mass were acquired both with and without scatter correction for 201Tl and 99mTc studies. Data were acquired on a newer generation gamma camera with an energy resolution of 8.7% at 140 keV. From conventional short-axis slices of the heart, circumferential count profiles were generated from five representative slices. Defect size was computed from the fraction of radians that fell below a fixed threshold value in each of the five count profiles. The nadir value (min/max) of the count profiles in each study was used as an index of image contrast. RESULTS: For both 201Tl and 99mTc, threshold values between 55%-60% gave the best correlation (r > 0.99), with the lowest average absolute error in estimating defect size (< 2.1%). Scatter correction reduced the average absolute error to 0.8% for 99mTc and 1.4% for 201Tl, significantly reduced the nadir values for both isotopes (p < 0.0001 for both 201Tl and 99mTc and led to a marked improvement in image quality for both tracers. CONCLUSION: Scatter correction reduces the error associated with measurement of infarct size, increases image contrast and improves image quality for both 201Tl and 99mTc, as assessed in a phantom model. PMID- 7472604 TI - Quantification of myocardial blood flow using dynamic nitrogen-13-ammonia PET studies and factor analysis of dynamic structures. AB - In this study, factor analysis of dynamic structures (FADS) was used to extract the "pure" blood-pool time-activity curves (TACs) and to generate parametric myocardial blood flow (MBF) images (pixel unit: ml/min/g). METHODS: Ten dynamic 13N-ammonia dog PET studies (three baseline, five hyperemia and two occlusion) were included. Three factors (TACs) and their corresponding factor images (the right ventricular and left ventricular blood pools and myocardial activities) were extracted from each study. The left ventricular factors matched well with the plasma TACs. The factor images of myocardium were then converted to a parametric images of MBF using a relationship derived from a two-compartment model. RESULTS: MBF estimates obtained from FADS correlated well with MBF estimates obtained with the two-compartment model (r = 0.98, slope = 0.84) and microsphere techniques (r = 0.96, slope = 0.94). FADS-generated MBF parametric images have better image quality and lower noise levels compared to those generated with Patlak graphical analysis. CONCLUSION: Regional MBF can be measured accurately and noninvasively with 13N-ammonia dynamic PET imaging and FADS. The method is simple, accurate and produces parametric images of MBF without requiring blood sampling and spillover correction. PMID- 7472605 TI - Simplified PET quantitation of myocardial glucose utilization. AB - The purpose of this study was to validate experimentally a simple method to quantify tissue glucose utilization with the brain reference index (BRI) using 14C-deoxyglucose and assess its clinical feasibility for myocardial PET. METHODS: To validate the BRI method, glucose utilization in myocardial and skeletal muscle was studied in rats with 14C-deoxyglucose after increasing doses of oral glucose loading. To assess clinical feasibility of the method, the BRI was applied to nine patients undergoing myocardial PET and compared to rMGU measured by the deoxyglucose model of Sokoloff et al. and by Patlak graphical analysis. The normal range of myocardial FDG uptake expressed as the BRI was estimated with four normal volunteers. RESULTS: In skeletal muscle, a dose-dependent increase of glucose utilization was observed during oral glucose loading with doses up to 4 mg/g. In the myocardium, glucose utilization increased with a glucose loading dose of up to 1 mg/g without increasing further at greater glucose doses. Ratios of maximal glucose utilization in glucose-loaded rats to 19-hr fasted rats (controls), expressed as the BRI for left and right ventricular myocardium and skeletal muscle were 4.16, 3.74 and 7.39, respectively. Glucose utilization of right ventricular myocardium was approximately 70% of left ventricular myocardium for all glucose-loaded conditions. For patients, the BRI correlated with rMGU; four of these patients had a constant plasma glucose concentration. CONCLUSION: Myocardial BRI is a sensitive indicator of rMGU that does not require dynamic data acquisition or constant plasma glucose concentrations. PMID- 7472606 TI - Effects of no flow and reperfusion on technetium-99m-Q12 kinetics. AB - The purpose of this study were to determine if 99mTc-Q12 tracer kinetics could be affected by the insult of total no flow followed by reflow and the effects of viability on clearance. METHODS: In six control buffer perfused rat hearts, flow was maintained at 12 ml/min throughout uptake and clearance. In six hearts (IR30), 30 min of no flow was followed by 60 min of reflow. In six hearts (IR60), 60 min of no flow was followed by 60 min of reflow. One millicurie 99mTc-Q12 was injected in control hearts or during reflow in the IR30 and IR60 hearts and myocardial clearance was monitored for 1 hr using a Nal detector. RESULTS: Control hearts demonstrated biphasic 99mTc-Q12 myocardial clearance with an early rapid clearance phase ending 5-10 min after injection (73.5% +/- 1.5% retention) followed by a late slow clearance phase (90.5% +/- 0.2% retention). IR30 hearts demonstrated a near identical clearance curve (74.3% +/- 0.9% early retention, 90.9% +/- 0.6% late retention). IR30 heart electron micrographs demonstrated predominantly ischemia insulted but viable cells. IR60 hearts also demonstrated a biphasic myocardial clearance, with a late slow phase similar to controls (91.9% +/- 0.6% retention). The early rapid phase was significantly faster than controls (61.1% +/- 3.4%). IR60 heart electron micrographs demonstrated predominantly injured nonviable cells. Well counting confirmed decreased retention in the IR60 rats compared to controls and IR30 rats. CONCLUSION: Technetium-99m-Q12 myocardial clearance is normally biphasic, with an early rapid phase ending after 5-10 min and a late slow phase. Ischemicaly insulted but viable myocardium created by 30 min of no flow followed by reflow has no effect on either clearance phase. This tracer warrants further study to determine its potential utility in assessing myocardial viability. PMID- 7472608 TI - Three-dimensional demarcation of perfusion zones corresponding to specific coronary arteries: application for automated interpretation of myocardial SPECT. AB - In this study, three-dimensional maps of specific coronary artery territories were derived and combined with normal distribution maps as a reference for automated characterization of defects, including location and size. METHODS: One hundred sixty-eight 99mTc-sestamibi myocardial perfusion SPECT scans from normal patients and patients with single-vessel disease were selected according to angiographic data. Five separate groups were established for men and women: normal, proximal left anterior descending (PLAD), distal left anterior descending (DLAD), right coronary artery (RCA) and left circumflex (LCx). All myocardial perfusion studies were aligned and sized to the same three-dimensional orientation using a previously developed automated image registration technique. Mean and variation three-dimensional templates were constructed from stress images in each group. Normal templates were demarcated with hypoperfusion regions obtained from disease templates. The defects were detected in the individual patient's images by a region-growing algorithm which identified abnormal voxels by comparison to the corresponding voxels in the mean and variation templates. RESULTS: Defects were quantified with respect to volume, location relative to the expected hypoperfusion zones and severity index. Abnormal regions could be marked directly on tomographic slices and visualized in various orientations. Single defects greater than 2% of the myocardium positioned within demarcated perfusion territories were detected in 105/119 abnormal patients and in 3/49 normal patients. CONCLUSION: Maps of myocardial perfusion zones created from images of angiographically selected patients provide a reference for automated localization of myocardial perfusion defects. A template-based region-growing is a robust technique for volumetric quantification and localization of abnormal regions. PMID- 7472607 TI - Rest myocardial perfusion/metabolism imaging using simultaneous dual-isotope acquisition SPECT with technetium-99m-MIBI/fluorine-18-FDG. AB - The purpose of this study was to develop a dual-isotope simultaneous acquisition (DISA) protocol using a multihead SPECT camera equipped with an ultrahigh-energy (UHE) collimator to evaluate simultaneously rest cardiac perfusion and metabolism with 99mTc-MIBI/18FDG. METHODS: Physical measurements were first performed with phantoms to develop the acquisition protocol. Fifteen patients underwent DISA SPECT with 99mTc-MIBI/18FDG to validate the protocol. To evaluate the quality of the 99mTc-MIBI images acquired with the UHE collimator, four patients underwent a resting 99mTc-MIBI scan acquired with a high-resolution, low-energy collimator prior to DISA-SPECT. RESULTS: With a window of 20% for both photopeaks and a 99mTc/18F concentration ratio of 3.2:1, the spillover from 18F into the 99mTc window is 6% of the counts in the window for normal subjects. Phantom images clearly demonstrated defects measuring 2 x 1 and 2 x 0.5 cm. Technetium-99m-MIBI images obtained with the UHE and high-resolution collimators provided similar diagnostic information. Using a stenosis of > 70% as criteria to diagnose coronary artery disease, DISA-SPECT had a sensitivity of 100% and a positive predictive value of 93%. CONCLUSION: Simultaneous evaluation of rest myocardial perfusion/metabolism with a multihead SPECT camera equipped with an UHE collimator is possible using 99mTc-MIBI/18FDG with a dual-isotope simultaneous acquisition protocol. PMID- 7472609 TI - Operator-less processing of myocardial perfusion SPECT studies. AB - We have developed a completely automated algorithm to generate reoriented tomographic images from projections in myocardial perfusion SPECT. METHODS: The algorithm consists of three software modules. The first module determines reconstruction limits for the projection dataset using two-dimensional feature extraction techniques. The second module reconstructs the projection images into transaxial images using standard filtered backprojection. The third module reorients the transaxial images into short-axis images. RESULTS: The algorithm was validated on 350 rest 201Tl and 350 stress 99mTc-sestamibi studies acquired on a single-detector (178 studies), a 90 degrees dual-detector (230 studies) or a triple-detector camera (292 studies). The complete processing sequence was successful in 93.6% of the studies (166/178 + 216/230 + 273/292). As for the individual modules, myocardial boundaries were correctly determined in 96.3% of the studies (171/178 + 222/230 + 281/292), while reorientation was successful in 97.2% of the studies (166/171 + 216/222 + 273/281). No significant difference in success rates for 201Tl versus 99mTc-sestamibi images was found. CONCLUSION: Our automated approach to myocardial perfusion SPECT processing is highly successful, intrinsically reproducible and can produce time and cost savings while improving accuracy in a clinical or research environment. PMID- 7472611 TI - Automatic quantification of ejection fraction from gated myocardial perfusion SPECT. AB - We have developed a completely automatic algorithm to quantitatively measure left ventricular ejection fraction (LVEF) from gated 99mTc-sestamibi myocardial perfusion SPECT images. METHODS: The algorithm operates in the three-dimensional space and uses gated short-axis image volumes. It segments the left ventricle (LV), estimates and displays endocardial and epicardial surfaces for all gating intervals in the cardiac cycle, calculates the relative left ventricular cavity volumes and derives the global EF from the end-diastolic and end-systolic volume, all without operator interaction. The algorithm for measuring LVEF was tested in 65 clinical patients undergoing 16-interval and 8-interval rest-gated SPECT and validated against first-pass radionuclide ventriculography. RESULTS: Automatic segmentation and contouring of the LV was successful in 65/65 (100%) of the studies. Agreement between EFs measured from 8-interval gated SPECT and EFs calculated from first-pass data was high (y = 2.44 + 1.03x, r = 0.909, p < 0.001, s.e.e. = 6.87). Agreement between EF values measured from 16-interval and 8 interval gated SPECT was excellent (y = -2.7 + 0.97x, r = 0.988, p < 0.001, s.e.e. = 2.65), the latter being on average lower by 3.71 percentage points. CONCLUSION: Our automatic method is rapid and highly agrees with conventional radionuclide measurements of EF, thus providing clinically useful additional information to complement myocardial perfusion studies. PMID- 7472610 TI - Cardiac studies with metaiodobenzylguanidine: a critique of methods and interpretation of results. PMID- 7472612 TI - Building organizational capacity to engage in research utilization. AB - The Orange County Research Utilization in Nursing (OCRUN) Project developed a regional network of nursing service organizations (NSOs) with a shared commitment to building organizational capacity to engage in research use. Over 3 years, 20 nursing service and 6 academic institutions participated in the OCRUN Project, sending 374 nurses to tiered OCRUN continuing education courses that focused on role-specific research utilization competency development. The project influenced nurse executives' perceptions of organizational readiness for change and catalyzed changes in strategic goals, accountability, and resources, ultimately influencing each NSO's reported research utilization processes and outcomes. Recommendations stemming from the OCRUN Project address potential NSO strategies to enhance research use. PMID- 7472613 TI - A cost analysis of a nurse-managed, voluntary community health clinic. AB - Nurse-managed, voluntary community health centers are thought to represent cost effective means of providing healthcare to the medically indigent. A cost analysis of a community healthcare clinic indicates that the cost of a clinic visit may be more expensive than the cost of a community physician visit when all costs (real and donated) are included. Increasing the number of patients seen per hour may be necessary to achieve more cost-effective delivery. PMID- 7472614 TI - Nurse information needs for efficient care continuity across patient units. AB - Continuity of care can be costly, unless information systems incorporate comprehensive patient data from all types of nursing units. This study identified nurses' communication processes and content needs when receiving patients in a large medical center averaging 3750 patient transfers monthly. A survey of 197 registered nurses in perioperative, intensive care, medical-surgical, outpatient, acute psychiatric, and long-term care settings revealed that some assessment content is considered important to all nurses, although the importance of other information can vary by specialty practice. Cost-benefit implications and planning applications are discussed. PMID- 7472615 TI - Productivity of nurse researchers employed in clinical settings. AB - With limited resources, there is increased scrutiny of the contributions of leadership staff, such as nurse researchers employed in the clinical setting (NRECS). The authors describe activities that pertain to research, presentations, publications, and procurement of funds for studies. Major obstacles that impede NRECS from achieving their goals, job performance evaluation and indices (staff, organization, and personal) that NRECS use to determine success in enacting their role also are presented. PMID- 7472616 TI - Reduced resource utilization through standardized outcome-focused care plans. AB - When evaluating ways to improve client outcomes, home healthcare administrators must consider the amount of resources used to provide the outcomes. Previous research has shown that outcome-focused care plans (OCPs) result in significantly better quality indicator scores than the traditional process-focused care plans (PCPs). The authors report the amount of resources used by a home healthcare agency when nursing care was guided by PCPs versus OCPs. The results showed that nurse visit time was less when OCPs were used. Outcome-focused care plans offer home healthcare administrators a mechanism to enhance quality while reducing resource utilization. PMID- 7472617 TI - Creating a climate to promote internal motivation in employees. PMID- 7472618 TI - Decentralized organizational structures and first-line nurse managers. PMID- 7472619 TI - Balancing judicial interests in HIV cases. Part 1, Issues for nurse administrators. AB - Legal issues involving the human immunodeficiency virus are consuming an increasing amount of the attention of hospital and nurse administrators. When the legal interests of patients, staff, and administrators conflict, healthcare administrators must determine which interest should prevail. Recently, two courts have dealt with this problem. The author discusses the first case- antidiscrimination versus workplace safety. She identifies the legal dilemma and suggests ways nurse administrators can recognize and reconcile such conflicts. PMID- 7472621 TI - Expert panel method for nurse staffing and resource management. AB - The Expert Panel Nurse Staffing and Resource Management Method is a bold new approach for the identification of nurse staffing requirements and the management of resources. Expert panels comprised of those most knowledgeable of the patient population and the uniqueness of specific patient care areas identify staff needed to meet clinical, administrative, education, continuous quality improvement, and research needs. In addition, the expert panels explore opportunities for systems improvements, work redesign, and administrative restructuring within the context of budgetary realities. PMID- 7472620 TI - Building a learning organization beyond the walls. AB - Senge's model of the learning organization served as the conceptual framework for the Vermont Nursing Initiative during its 5-year effort to create change across Vermont's 15 community hospitals. The author describes the process of investing in the intellectual capital of these hospital nurses through the development of Senge's five disciplines. The unexpected outcome of this process was the development of a nursing learning organization that reaches beyond hospital walls. PMID- 7472622 TI - The season of change. AB - The American Organization of Nurse Executives (AONE) is the national organization of nurses in executive practice. AONE provides leadership, professional development, advocacy, and research to advance nursing practice and patient care, promote nursing leadership excellence, and shape healthcare public policy. This column, written by officers and executive staff members of AONE, raises issues and discusses trends of importance to nursing administrators. PMID- 7472623 TI - Nurse managers and their future in New South Wales. AB - The goal of International Perspectives is to share the views of nurse leaders, from countries other than the United States, on global healthcare issues. The focus is on identifying healthcare management problems and their implications, research priorities, policy development, and ethicalmoral dilemmas faced by nurse administrators. You are encouraged to send the names and addresses of international nurse leaders who might be interested in participating, as well as your ideas, for format and content of the department. PMID- 7472624 TI - Organizational culture and ethics work satisfaction. AB - Nurse executives have opportunities for creating constructive organizational cultures and for supporting ethical healthcare practice. Because culture and ethics are so central to effective nursing practice, this study explored relationships between them, measuring ethics as work satisfaction pertaining to ethics. This article provides initial assessment of one dimension of ethics, ethics work satisfaction, and examines its relationship to organizational culture. PMID- 7472625 TI - Peer social support: a key to care giver morale and satisfaction. AB - Social support is a group dynamic, but studies have not identified specific strategies for managers to use as direct environmental resources. The authors discuss a study that explored group social support as a component of job morale for bedside care givers. Specific unit-based factors are identified for managers to develop focused staff social support activities. These include means of universal recognition, use of common space, methods to develop team culture and styles, and methods of communication. PMID- 7472626 TI - Keeping the care in nursing care: our biggest challenge. AB - Although caring is the essence of nursing practice, many nurses are concerned that their ability to care is being compromised by the tumultuous changes occurring in the current healthcare environment. The author explores the challenge of caring in nursing practice, reviews what recent research has shown us about caring in nursing and identifies what we as nurse executives can do to enhance our own caring skills and the caring expertise of clinical nurse colleagues. PMID- 7472627 TI - Patient care coordinators: successfully merging utilization management and discharge planning. AB - Facilitating patient-focused, cost-effective care throughout the continuum is a challenge that requires creativity of healthcare administrators. At Children's Hospital, Boston, the patient care coordinator (PCC) role was developed to improve communication and linkage among healthcare providers, payer organizations, and families by blending the functions of utilization management and discharge planning into a unit-based leadership role. The authors describe the PCC role and illustrate its goals and outcomes with examples. PMID- 7472628 TI - Home health quality outcomes. Fee-for-service versus health maintenance organization enrollees. AB - Quality outcomes were compared between home health patients enrolled in the traditional Medicare fee-for-service (FFS) program versus a health maintenance organization (HMO) with a Medicare cost contract with the federal government. The quality outcome scores were similar between the two patient groups. However, other home health research showed superior quality outcomes for patients enrolled in the traditional Medicare FFS program versus an HMO with a risk contract with the federal government. Before signing contracts with HMOs, home health administrators will want to identify the type of Medicare contract the HMO has with the federal government. PMID- 7472630 TI - Advanced practice nursing: the winds of change. PMID- 7472629 TI - The cost-effectiveness of a special care unit to care for the chronically critically ill. AB - To assess the relative value of healthcare programs, technologic innovations, and clinical decisions, policymakers are searching for ways to evaluate cost effectiveness. What constitutes cost-effectiveness and how should it be measured? The authors discuss ways in which the cost-effectiveness of clinical programs can be measured and describes various methods of assessing both costs and effectiveness. Comparison of the cost-effectiveness of a nurse managed special care unit with that of traditional intensive care units illustrates some of these methods. PMID- 7472631 TI - Comfort at the end of life: palliative care policy. AB - Despite the lack of evidence for improvement in symptom relief, quality of life, or length of survival, patients who are terminally ill often are subjected to unnecessary invasive procedures and denied the symptom relief that modern technology and pharmacology make possible. Nursing administrators are in an ideal position to initiate interdisciplinary policy changes in this area and are encouraged to become familiar with the principles of palliative care. A recently developed policy is described and a review of supportive literature is given. PMID- 7472632 TI - Coaching practices of first-line nurse managers in hospital wards with highly satisfied personnel. PMID- 7472633 TI - Unlicensed assistive personnel: legal considerations. PMID- 7472634 TI - Balancing judicial interests in HIV cases: Part 2, Issues for nurse administrators. AB - Legal issues involving human immunodeficiency virus are consuming an increasing amount of the attention of hospitals and nurse administrators. At times, the legal interests of patients, staff, and administrators conflict, and it is the duty of healthcare administrators to determine which interests should prevail. Recently, two courts have dealt with this problem. In a previous article (JONA October 1995), the author discussed the first case, which involved antidiscrimination versus workplace safety. In this article, the author discusses the second case, which also involves issues of antidiscrimination and workplace safety, but with a decidedly different twist. She identifies the legal dilemma and suggests ways in which nurse administrators can recognize and reconcile such conflicts. PMID- 7472635 TI - Changing trends in healthcare: implications for baccalaureate education, practice and employment. AB - The authors surveyed 96 hospitals and agencies in Massachusetts to determine: 1) the effect of the changes in health care delivery on RN utilization, particularly baccalaureate nurses, and 2) to identify educational changes necessary to prepare baccalaureate nursing graduates for employment and practice in the future. An analysis of the results of the survey of the 50 nurse executives who responded to the study identifies the projected needs and requirements of the service sector for delivering nursing care, and translates them into suggestions for revisions of the baccalaureate curriculum. PMID- 7472636 TI - Improving examination performance using Exam Analysis. AB - The Exam Analysis is a diagnostic and prescriptive program in which a student and an instructor identify why the student failed to answer specific questions correctly on an examination. Problems related to incorrect answers are then categorized into five problem areas. This analysis is used to develop a plan which will help the student utilize specific skills in order to improve examination performance. This article describes the Exam Analysis program, gives specific information which will enable an instructor to assist a student in completing an Exam Analysis, and discusses the results of a survey in which 105 nursing students evaluated the impact of participating in the program on subsequent examination performance. A large majority of the students surveyed agreed that participation in the Exam Analysis program resulted in improved examination performance. Improved exam-taking skills, better study skills, decreased test anxiety and increased self-confidence were identified as Exam Analysis program outcomes by the students included in the study. PMID- 7472637 TI - Pain management instruction in nursing curricula. AB - The persistent problem of inadequate assessment of pain, undermedication for pain by nurses, and reports of nurses themselves that they are unprepared to manage various types of pain provided direction for this study. The purpose was to investigate time allocated to pain content, the nature of the theoretical content and clinical experiences in nursing curricula, and faculty satisfaction with their graduates' preparation for pain management in baccalaureate and associate degree programs. The 80% response rate included 177 associate degree and 174 baccalaureate nursing programs. No significant differences between associate degree and baccalaureate programs for the amount of time allocated to pain content, pharmacological management of pain and nonpharmacological pain relief methods were found. Nonpharmacological methods are introduced to students, but most programs do not provide in-depth instruction. The majority of respondents indicated they were satisfied with their graduates' preparation for pain management, but less satisfied with the amount of time allocated to pain content in the curriculum. Implications for education and research are discussed. PMID- 7472638 TI - A research development program for minority honors students. AB - This article describes an experiential undergraduate honors research program for minority students. Through a participatory and mentoring process students are able to complete a research project, present it at a national colloquium, and prepare it for publication in a 2-year program. A review of the literature on different methods to teach research was presented. The outcome of the project suggests the model was highly successful in achieving its goals. PMID- 7472639 TI - Transcultural mentoring: an experience in perspective transformation. AB - This article describes the phases of a transcultural mentoring relationship between Chinese nurses and Canadian nursing professors. Mezirow's adult learning theory of perspective transformation was used to guide this educational experience. An understanding of this learning process sensitized the mentors to the disorienting dilemma and culturally induced behaviors of their proteges. The aims of this article are to provide a description of the mentoring process using Mezirow's framework, describe how this process differed from the traditional mentoring relationship, identify the major factors that influenced the relationship, describe what was learned from the experience and to make recommendations based on the authors' experiences. PMID- 7472640 TI - Barriers and breakthroughs: substance abuse curricula in nursing education. PMID- 7472641 TI - Academic and demographic variables related to completion status of nursing students in master's degree programs. PMID- 7472642 TI - Evaluation of an educational program to prepare neonatal nurse practitioners. PMID- 7472643 TI - Proverb interpretation to assess mental status. PMID- 7472644 TI - Promoting healthy parenting through prenatal education. PMID- 7472645 TI - Processes of care. Comparisons of certified nurse-midwives and obstetricians. AB - Prenatal and intrapartum care provided to 1,181 women, all meeting risk requirements for nurse-midwifery care, by certified nurse-midwives (n = 471) and obstetricians (n = 710) are compared using indicators of physical and of educational/psychosocial components of maternity care. Data are from clinical records and questionnaires completed by the women. Bivariate analyses show that the two provider groups differ on some, but not all, processes of care. When the woman's evolving health status, personal characteristics, and preferences are controlled, there are significant differences that confirm two models of care. The nurse-midwifery approach emphasizes educational/psychosocial care and restrained, individualized use of technology. The obstetrics approach emphasizes more routine use of state-of-the-art technology. This study contributes new information to substantiate different models but also shows that both provider groups use elements of both. The difference in emphasis should encourage collaborative practice, given the shared basis for maternity care, whether it is provided by certified nurse-midwives or obstetricians. PMID- 7472646 TI - The essence of nurse-midwifery care. The woman's story. AB - The purpose of this qualitative research study was to discover the experience of the woman cared for by a nurse-midwife. Six variously situated and ethnically diverse women were interviewed about their experience with nurse-midwifery care. Using Colaizzi's phenomenological method, the data were analyzed extracting 151 significant statements that were clustered into nine themes. A picture of the essential structure of the women's experience of nurse-midwifery care unfolded and was returned to them for their comments and confirmation. Women talked about a relationship built on respect, trust, and alliance. It was the nurse-midwife herself, her qualities and behaviors, her concern and caring, and her respect for women that laid the foundation for how the women in this study perceived the experience. Repeatedly, women articulated the respect for their time, their families, their fears, and their need for information. Ultimately, it was this respect from the nurse-midwife that empowered the women to determine and direct their care. PMID- 7472648 TI - Delayed reporting of sexual assault. Implications for counseling. AB - The nurse-midwife may be called upon to counsel the victim of a sexual assault many months or years after the crime. It is important to assess which of these women need referral to a licensed therapist and which can benefit from empathy and skilled counseling techniques that nurse-midwives can add to their repertoire of expertise. PMID- 7472647 TI - Weight change in the postpartum period. A review of the literature. AB - Women who gain excessive weight during pregnancy are at risk for postpartum weight retention and long-term obesity. Familiarity with current research related to postpartum weight loss can aid the nurse-midwife in providing sound, clinical advice to women at risk for postpartum weight retention. This article provides a review of current literature and research studies related to postpartum weight loss, weight retention, and weight gain. An overview of gestational weight gain and fetal growth, recommendations of the Institute of Medicine, complications of maternal obesity, postpartum weight-loss patterns, and factors that affect postpartum weight loss are discussed. PMID- 7472649 TI - The transition from lay midwife to certified nurse-midwife in the United States. AB - This paper presents the results of a survey that was conducted between November 1992 and March 1993. Its target group consisted of midwives in the United States who began their careers as lay midwives and later decided to become certified nurse-midwives (CNMs). Questions address their demographic and socioeconomic characteristics to compile a profile that can be compared with other midwife populations. The survey elicited information regarding what motivated the decisions to change status and how these decisions have made an impact upon respondents' personal lives, family life styles, and income; identified how becoming a nurse-midwife changed the respondents' work in regard to their own community, the clientele they serve, and their style and site of practice, and how the change in identity affected their self-perceptions as midwives as well as their relationship to the health care system, the legal system, and the established midwifery system; it investigated how respondents viewed their preparation for midwifery and how they felt about the relationship of nursing to midwifery; and, it addressed how these midwives related to, were influenced by, and influenced the two major professional midwifery associations in the United States, the Midwives Alliance of North America and the American College of Nurse Midwives. The results showed that, overall, the respondents valued their lay midwifery background, felt positive about their CNM educational programs, and showed a preference for direct entry and apprenticeship programs in which the RN is not a prerequisite. PMID- 7472651 TI - Liver glucose-6 phosphatase activity is inhibited by refeeding in rats. AB - This study was conducted to determine whether inhibition of hepatic glucose-6 phosphatase is involved in the mechanism of suppression of hepatic glucose production during the postprandial period. We studied the time course of changes in the enzyme activity by refeeding food-deprived rats with nonpurified diet. The Vmax of the enzyme, assayed in homogenates from livers freeze-clamped in situ in anesthetized 48-h unfed rats (12.3 +/- 0.15 U/g wet liver, mean +/- SEM, n = 6) was progressively decreased upon refeeding: 11.1 +/- 0.5, 8.5 +/- 0.4 and 7.9 +/- 0.5 U/g, in rats refed for 90, 180 (P < 0.01) and 360 min (P < 0.01), respectively. The Km of the enzyme was not affected by refeeding. No inhibition of the enzyme was observed in microsomes purified from these homogenates, suggesting a metabolite-induced inhibition mechanism. To assess the role of insulin in the inhibition, we assayed the glucose-6 phosphatase activity in similarly processed liver homogenates from food-deprived rats perfused with insulin at physiological and supraphysiological concentrations, whereas plasma glucose was maintained at the basal level by adapted glucose perfusion (euglycemic clamps). No inhibition of glucose-6 phosphatase was found under these conditions, suggesting that insulin cannot by itself account for the inhibition observed in the refeeding experiments. These data constitute the first demonstration of the inhibition of glucose-6 phosphatase activity during the postprandial period. PMID- 7472650 TI - Midwifery in Germany. Its past and present. AB - This article introduces midwifery in Germany to the North American midwife. The development of the midwifery profession since the 12th century is presented. Some regulations that govern midwifery practice today were initiated by the clergy, city government, and physicians during past centuries. Present-day midwifery training and professional opportunities are described. The organization and peculiarities of the German health care system and some effects of German reunification that are relevant to midwifery practice are also discussed. Midwives work in several areas and are regulated by federal and state laws which support and secure their practice. The promotion of midwifery interests frequently encounters resistance by obstetricians. There is a long-standing conflict related to an overlap of skills and services that both professions claim as their own. In recent years, more women have demanded and used midwifery services. In 1992, 811,774 deliveries were performed by midwives in 1,176 German hospitals with varying styles of labor management. This variety ranged from university hospitals, which emphasized modern technologies, to institutions that supported a comprehensive midwifery approach. The author challenges the classical practice of midwifery and contrasts it with a delivery that incorporates several features in a comprehensive approach. PMID- 7472652 TI - Dietary methionine does not reduce penetrance in curly tail mice but causes a phenotype-specific decrease in embryonic growth. AB - The mouse mutation, curly tail, has incomplete penetrance and variable expression. Approximately 60% of the mice have a curly tail (CT), from which up to 20% may have lumbosacral spina bifida. Approximately 40% are normal, with a straight tail (ST). We tested whether L-methionine, which reduces the penetrance of neural tube defects in the Axd mouse mutant, has beneficial effects in the curly tail mutant. A single injection of L-methionine (200-1600 mg/kg body wt) on d 9 of pregnancy had no effect on the embryos, whereas there was a minor increase in penetrance at the highest dose. Chronic supplementation of L-methionine via the drinking water (1554 mg.kg body wt-1.d-1) did not shift penetrance. However, it decreased the weight of d 13 embryos from ST dams but not of those from CT dams. This phenotype-specific difference in response was evident and most unexpected. Mice from curly tail and other inbred strains were subjected to an L methionine loading test and serum homocysteine assay. The different strains varied in their basal serum homocysteine concentrations, and they had proportionate significant increases after L-methionine loading. In CT and ST mice, basal serum homocysteine concentrations as well as the levels after loading were similar to each other and intermediate in the range of the mice tested. We conclude that L-methionine does not reduce penetrance in the curly tail mouse and that this strain reflects no derangement in L-methionine handling. PMID- 7472653 TI - Nutritional modulation of guinea pig skin hyperproliferation by essential fatty acid deficiency is associated with selective down regulation of protein kinase C beta. AB - In a previous study we demonstrated that 13-hydroxyoctadecadienoic acid (13 HODE), a 15-lipoxygenase metabolite of linoleic acid is incorporated into epidermal phosphatidyl 4,5-bisphosphate (PtdIns 4,5-P2) and released as 13-HODE containing-diacylglycerol (13-HODE-DAG). In vitro, 13-HODE-DAG was shown to selectively inhibit epidermal total protein kinase C (PKC-beta) activity. To determine whether these observations are relevant in vivo, guinea pigs were made essential fatty acid deficient (EFAD) by feeding them a basal diet supplemented with 4% hydrogenated coconut oil for 8 wk. Tissue levels of putative 13-HODE-DAG, protein kinase C (PKC) isozymes and tissue hyperproliferation were determined in the epidermal preparations from skin of control safflower oil-fed guinea pigs, those fed EFAD diet and those fed EFAD diet followed by the control diet for 2 wk. Our data revealed that cutaneous 13-HODE and 13-HODE-DAG were significantly lower in EFAD animals than in safflower-fed controls. These reductions were associated with both elevated epidermal hyperproliferation and elevated expressions and activities of PKC-alpha and beta-isozymes. Refeeding the animals with safflower oil for 2 wk replenished tissue levels of 13-HODE-DAG, which inversely correlated with the selective down regulation of PKC-beta expression and activity and the reversal of hyperproliferation. In contrast, although, the expression and activity of PKC-alpha was elevated in the epidermis of the EFAD guinea pigs, this elevated PKC-alpha expression was not down regulated after refeeding the safflower oil diet to the animals.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7472654 TI - Pyridoxine and pyridoxine-5'-beta-D-glucoside exert different effects on tissue B 6 vitamers but similar effects on beta-glucosidase activity in rats. AB - Pyridoxine glucoside is a partially available form of vitamin B-6 present in plant-derived foods. In this set of three studies, rats were fed diets containing different concentrations of either pyridoxine or pyridoxine glucoside in the presence or absence of a fixed concentration of pyridoxine for 2 wk. The distribution of B-6 vitamers and beta-glucosidase activity in tissues was examined to determine the metabolic effects of chronic consumption of pyridoxine glucoside. Rats fed pyridoxine glucoside either with or without pyridoxine exhibited a significant increase in the amount of hepatic pyridoxine 5'-phosphate (not detected in rats fed pyridoxine alone), whereas hepatic pyridoxal 5' phosphate was decreased with increasing dietary pyridoxine glucoside. The activity of cytosolic beta-glucosidases in small intestine and kidney was affected by the dietary concentration of both pyridoxine and pyridoxine glucoside. Enzymatic activity capable of hydrolyzing pyridoxine glucoside was found in mucosal and intraluminal fractions of small intestine and in the kidney. Other tissues examined, including liver, spleen and stomach, did not hydrolyze pyridoxine glucoside in detectable quantities. These findings indicate that microbial and mucosal enzymes can participate in the intestinal hydrolysis of pyridoxine glucoside and that the kidney may contribute to postabsorptive hydrolysis. These findings further support the observations that dietary pyridoxine glucoside influences vitamin B-6 metabolism. PMID- 7472655 TI - Geraniol, an inhibitor of mevalonate biosynthesis, suppresses the growth of hepatomas and melanomas transplanted to rats and mice. AB - Farnesyl-pyrophosphate is required for the posttranslational modification of G proteins including p21 ras, prelamin A and lamin B, each of which plays an essential role in cell proliferation. As a consequence, competitive inhibitors of mevalonate synthesis, the rate-limiting substrate for the synthesis of the prenyl pyrophosphates, arrest cultured cells at the G1/S interface of the cell cycle and initiate apoptotic cell death. Geraniol, an acyclic monoterpenoid alcohol, suppresses 3-hydroxy-3-methylglutaryl coenzyme A (HMG-CoA) reductase activity and concomitantly arrests the growth of cultured tumor cells. We evaluated the impact of dietary geraniol on the growth of two tumors. In the first study, geraniol (23 mmol/kg diet, 350 mumol/d) was fed to male buffalo rats for 14 d before and for 42 d after the transplant of Morris 7777 hepatomas. Tumor growth was suppressed (P < 0.001). In the second study, the dose-dependent impact of geraniol on the growth of B16 melanomas was assessed. Dietary geraniol (0.65, 6.5 and 65 mmol/kg diet) was fed to female C57BL mice for 14 d before and for 21 d after tumor transplant. Tumor growth was suppressed (P < 0.02) by 6.5 and 65 mmol geraniol/kg diet. PMID- 7472656 TI - Insulin treatment affects brain protein synthesis rate in streptozotocin-induced diabetic rats. AB - The purpose of this study was to determine whether insulin treatment affected the rate of brain protein synthesis in diabetic rats. Experiments were conducted on three groups of rats given streptozotocin without insulin treatment (diabetes), treated with streptozotocin + insulin (diabetes + insulin) or treated with neither streptozotocin nor insulin (control). The fractional rates of protein synthesis in brain, liver and kidney of diabetic rats treated with insulin were significantly greater than in diabetic rats without insulin treatment. In brain, liver and kidney, the RNA activity [g protein synthesized/(g RNA.d)] was significantly correlated with the fractional rate of protein synthesis. The RNA concentration (mg RNA/g protein) was not related to the fractional rate of protein synthesis in any organ. The results suggest that insulin treatment of diabetic rats is likely to increase the rate of protein synthesis in the brain, and that RNA activity is related to the fractional rate of brain protein synthesis. PMID- 7472657 TI - Combined growth hormone (GH) and insulin-like growth factor-I (IGF-I) treatment is more effective than GH or IGF-I alone at enhancing recovery from neonatal malnutrition in rats. AB - The effect of hormonal therapy on enhancing recovery from neonatal malnutrition was assessed in rats. Malnutrition was induced by maternal food restriction (60% control intake) during lactation. On d 16 postpartum, restricted pups were refed by cross-fostering to control dams and were subcutaneously administered growth hormone (GH), insulin-like growth factor-I (IGF-I), GH + IGF-I or saline. At d 21, pups were weaned and continued hormonal treatments until d 39 postpartum. By d 39, body weight of GH (96% control) and GH + IGF-I- (98%) treated animals were normal, whereas IGF-I (88%) and saline- (85%) treated animals were still stunted. Hormone effectiveness was age dependent, with growth rates of GH + IGF-I-, IGF-I- and GH-treated pups being greatest between d 16 and 30, d 16 and 21 and after d 30, respectively. Protein accretion by d 39 was higher (P < 0.01) in GH and GH + IGF-I groups than saline- or IGF-I-treated groups. On d 39, serum IGF-I concentrations were as follows: GH + IGF-I > IGF-I > GH = saline (placebo), indicating that an elevated serum IGF-I was not required for GH-stimulated growth. Of the three hormonal treatments, GH + IGF-I therapy was most efficacious at promoting rapid and complete body weight recovery and supporting lean body mass accretion. PMID- 7472658 TI - Delaying the introduction of complementary food until 6 months does not affect appetite or mother's report of food acceptance of breast-fed infants from 6 to 12 months in a low income, Honduran population. AB - Low income, primiparous mothers who had exclusively breast-fed for 4 mo were randomly assigned to one of three groups: 1) continued exclusive breast-feeding to 6 mo (EBF), 2) introduction of complementary foods at 4 mo, with ad libitum nursing 4-6 mo (SF), and 3) introduction of complementary foods at 4 mo, with maintenance of base-line nursing frequency 4-6 mo (SF-M). After the intervention phase (4-6 mo; n= 141), home visits were conducted for a subsample at 9 (n = 60) and 12 (n = 123) mo. At each visit, an observer recorded infant food intake at the midday meal and interviewed the mother regarding usual feeding patterns and the infant's acceptance of 20 common food items. All but two infants (1.5%) were breast-fed to 9 mo and all but eight (6%) to 12 mo. There were no significant differences among groups in breast-feeding frequency, amount or number of foods consumed at the midday meal, percentage of food offered that was consumed, usual daily number of meals and snacks, number of food groups consumed, or overall food acceptance score. Frequency of consumption of foods from eight different food groups (dairy, meats, eggs, grains, beans, fruits, vegetables, tubers) was not significantly different among groups except that, at 9 mo only, the SF group (but not the SF-M group) consumed more vegetables than did the EBF group. These results indicate that delaying the introduction of complementary foods until 6 mo does not adversely affect appetite or food acceptance among breast-fed infants. PMID- 7472659 TI - Validation of the Radimer/Cornell measures of hunger and food insecurity. AB - Hunger and food insecurity have been identified as core indicators of an individual's nutritional state that should be assessed in nutrition surveillance activities. Such an assessment requires a valid measure of these phenomena. This paper describes further work on the construction of measures of hunger and food insecurity based on the Radimer/Cornell items and provides an assessment of their validity. A random sample survey of 193 households with women and children living at home was conducted in 1993 in a rural county in New York State. A questionnaire containing the Radimer/Cornell hunger and food insecurity items, information about demographic characteristics, frequency of fruit and vegetable consumption, and household food supplies was administered to subjects. Measures were constructed that identified households experiencing household- and individual-level food insecurity and households with hungry children. The construct and criterion-related validity of the measures was assessed. In relation to criterion-related validity, as food insecurity worsened, there was a significant and progressive increase in the percentage of subjects participating in food programs and having low income, education and employment and a significant decline in average household food availability and fruit and vegetable consumption. These results support the validity of the Radimer/Cornell measures and illustrate their ability to differentiate among groups of households experiencing increasingly severe food insecurity and hunger. PMID- 7472660 TI - A meta-analysis of the effect of calcium intake on bone mass in young and middle aged females and males. AB - Evidence of a direct relationship between dietary calcium and bone mass in young adults is inconsistent. Therefore, a meta-analysis was performed to evaluate the existing literature concerning this relation in premenopausal women and adult men between the ages of 18 and 50. A computer-aided search of published papers from 1966 through 1994 was conducted. The quality of each study was assessed by examining the study design. The results of the well-designed studies wer combined to obtain quantitative conclusions. A total of 33 eligible studies were identified in the literature, representing 27 cross-sectional studies, two longitudinal and four intervention studies. The results of the cross-sectional studies in permenopausal females showed a significant correlation coefficient (r = 0.13, 95%-confidence interval = 0.09-0.16) as well as partial correlation coefficient (partial r = 0.08, 95%-confidence interval = 0.05-0.12), both weighted by sample size. The intervention studies found that calcium supplementation of approximately 1000 mg/d in premenopausal women can prevent the loss of 1% of bone/y at all bone site except in the ulna. In conclusion, the studies published to date seem to offer overall evidence that calcium intake is positively associated with bone mass in premenopausal females. This association is fairly consistent across the different study designs and is strengthened by the fact that the results are based only on studies with a high methodological quality. In males, too few studies (only three) were published to draw firm conclusions. PMID- 7472661 TI - Short-term changes in calcium but not protein intake alter the rate of bone resorption in healthy subjects as assessed by urinary pyridinium cross-link excretion. AB - This study was conducted to determine whether the markers of bone resorption, pyridinium cross-links of collagen, are sensitive to changes in dietary protein and calcium intake. Fifteen young healthy subjects (7 males and 8 females) participated in three 5-d diet periods. Dietary intake during each dietary period consisted of: 1) low nitrogen and low calcium [0.49 +/- 0.11 g protein/ (kg.d), 429 +/- 190 mg calcium/d]; 2) low nitrogen and high calcium [0.44 +/- 0.08 g protein/(kg.d), 1643 +/- 171 mg calcium/d]; and 3) a high nitrogen and high calcium [2.71 +/- 0.75 g protein/(kg.d), 1589 +/- 633 mg calcium/d] diet, and this was compared with subjects' baseline dietary intake [0.99 +/- 0.51 g protein/(kg.d), 589 +/- 152 mg calcium/d]. The order of these diets was randomly assigned. Twenty-four-hour and 3-h urine samples were collected before and during each dietary period and were analyzed for pyridinium cross-links (pyridinoline, deoxypyridinoline), nitrogen and creatinine. The rate of pyrdinium cross-link excretion did not vary with protein intake but was approximately 33% lower (P < 0.01) during periods of high compared with low calcium intake. These data indicate that a short-term increase in calcium intake is accompanied by a reduced rate of bone resorption and that this effect is independent of dietary protein intake. PMID- 7472662 TI - Some aspects of neonatal essential fatty acid status are altered by linoleic acid supplementation of women during pregnancy. AB - To study the effect of maternal linoleic acid [18:2(n-6), LA] supplementation during pregnancy on neonatal essential fatty acid status, pregnant women with relatively low plasma linoleic acid concentrations before 16 wk of gestation (n = 21) were supplied with foods rich in linoleic acid, resulting in an additional intake of 10 g/d of linoleic acid from the 20th week of gestation until delivery. One of the two control groups consisted of pregnant women with comparably low plasma linoleic acid concentrations at the start of the study (LL-control group, n = 22); the other consisted of women with habitually high plasma linoleic acid concentrations (HL-control group, n = 21). The neonatal essential fatty acid status was assessed by determining the fatty acid composition of phospholipids (PL) isolated from umbilical plasma and umbilical vessel walls. The maternal linoleic acid status in the LA-supplemented group increased to a level comparable to that of the HL-control group, but the neonatal linoleic acid status did not differ from that of either control group. Linoleic acid supplementation did result in slightly, but significantly, higher total amounts of (n-6) long-chain polyenes in umbilical plasma and vein vessel wall phospholipids compared with the LL-control group. This increase was associated with significantly lower total amounts of (n-3) long-chain polyenes. In the HL-control group, the concentration of (n-3) long-chain polyenes in umbilical plasma and vessel walls was significantly lower than in the LA-supplemented and the LL-control group.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7472663 TI - Dietary soybean protein decreases plasma taurine in cats. AB - Commercial dry and canned diets fed to cats cause approximately two- and fourfold increase in the taurine requirement, respectively, as compared with that observed for purified diets. In two experiments, the effect of source and level of protein and fiber in the diet on the concentration of taurine in plasma and whole blood of cats was studied. All diets contained 1 g taurine/kg dry matter. When a casein based diet containing either 25% or 50% protein was given to cats for 6 wk, no difference in plasma taurine concentration was observed; however, substituting soybean protein for casein resulted in a significant (P < 0.01) decrease in plasma taurine concentration of cats in the 50% soybean protein group, but not in the 25% soybean protein group. In Experiment 2, the food intake of cats was limited [26 g dry matter/(kg body weight.d)], and the protein was 30 or 60% of the diet. Cats fed 60% soybean protein or casein diets had significantly lower plasma taurine concentrations than cats fed a 30% casein diet, with the 60% soybean protein diet causing the greater decrease. There was no effect of either 2 or 4% soybean fiber on plasma taurine concentration as compared with the same diet without the added fiber. The taurine concentration in plasma was higher (P < 0.05) in male cats than in female cats. Protein source, amount in the diet and gender did not affect the whole blood taurine concentration. Cats given diets containing 60% casein or soybean protein diets excreted a greater amount of fecal total bile acid and total taurine than cats given a 30% casein diet.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7472664 TI - Diet-dependent and diet-independent metabolic responses underlie growth stasis of pigs at weaning. AB - The weaning transition in domestic animals involves profound environmental and nutritional changes. Growth stasis is commonly observed in pigs during this period, resulting in significant losses to the swine industry. It has been suggested that the reduced growth rate reflects immune sensitivity to soy antigens in commercial diets; however, few studies have defined metabolic responses in pigs weaned to diets containing soybean meal. The impact of environmental and social changes at weaning on physiological functions, i.e., the metabolic adjustment to new nutritional substrates, has also been overlooked. Accordingly, the aim of this study was to distinguish diet-dependent and diet independent metabolic responses in pigs weaned to a commercially available corn soy diet. Focus was on the endocrine pancreas and cytokines associated with stress responses. Three-week-old crossbred pigs were weaned to a corn-soy (cereal) or a milk-based (milk) diet. Blood samples taken 0 (weaning), 1, 2, 5 and 7 d post-weaning demonstrated diet-independent responses including increased (P < 0.05) plasma glucagon concentrations, decreased (P < 0.05) glucose concentrations, increased (P < 0.05) interleukin-1 (IL-1) concentrations during the first 2 d post-weaning, and increased (P < 0.05) fibrinogen concentrations during the latter part of the study. In response to dietary treatment, milk-fed pigs had higher (P < 0.05) plasma insulin and glucose concentrations, reflective of higher food intake during the early post-weaning period. This clear distinction between diet-dependent and diet-independent metabolic responses at weaning suggests consideration of novel strategies to overcome the characteristic weanling growth stasis in pigs. PMID- 7472665 TI - Methylmalonic acid inhibits respiration in rat liver mitochondria. AB - Methylmalonic acid (MMA), which accumulates and is excreted in urine in mammals during vitamin B-12 deficiency, has been reported to inhibit succinate dehydrogenase, an enzyme involved in the mitochondrial tricarboxylic acid (TCA) cycle in rat liver. The enzyme inhibition by MMA may lead to various metabolic disorders as well as inhibition of mitochondrial energy generation in vitamin B 12-deficient mammals. To clarify the inhibition of succinate dehydrogenase by MMA in intact rat liver mitochondria, the effect of MMA on mitochondrial respiration was studied. When 6 mmol/L MMA was added to the reaction mixture for measuring mitochondrial respiration with succinate as a substrate, MMA was taken up and accumulated by the mitochondria (34-53 mmol/L). The accumulation of mitochondrial MMA was stimulated by the addition of ADP. Methylmalonic acid competitively inhibited State 3 mitochondrial respiration, and the Ki for the acid was 4.2 +/- 0.4 mmol/L. Although the respiratory control ratio decreased with increasing MMA concentration, the acid did not affect the phosphorus/oxygen ratio. Mitochondrial MMA accumulation secondary to vitamin B-12 deficiency inhibits succinate dehydrogenase and may contribute to various metabolic disorders associated with vitamin B-12 deficiency. PMID- 7472666 TI - Choline distribution and metabolism in pregnant rats and fetuses are influenced by the choline content of the maternal diet. AB - Choline supplementation of pregnant rats between d 12 and 17 of pregnancy permanently enhances the spatial memory of offspring; however, the mechanism is unknown. We examined the effect of choline supplementation on metabolism of orally ingested choline by nonmated rats and pregnant rats and their fetuses. We studied the metabolism of an acute oral dose of 14C-choline chloride in pregnant and nonmated rats with and without choline supplementation (25 mmol/L choline chloride in water) on d 12-17 of pregnancy. During the first 2 h after oral dosing, plasma radiolabeled choline was detectable, whereas plasma choline metabolites contributed little to total radioactivity at any time. The pattern of accumulation of label in placentas was similar in all groups. Fetal tissues (i.e., brain, liver and carcass remnant) contained primarily 14C phosphatidylcholine and 14C-phosphorylcholine. Also, we examined the fetal tissue distribution of isotopically labeled (deuterated) choline derived from the diet and from the dietary choline supplement. The distribution patterns for radiolabeled choline metabolites in fetuses of supplemented dams accumulated significantly (P < 0.01) more of their total choline and its metabolites than fetuses of control dams during d 12-17 of gestation (50 vs. 20%). In fetuses from supplemented dams, betaine concentrations were greater than in fetuses from control dams in all organs assayed (by 36-57%). Phosphorylcholine concentrations in brain of fetuses from supplemented dams were also greater. These experiments identify potential metabolites of choline that might mediate the observed effects on brain development in the rats. PMID- 7472667 TI - Developmental changes of free amino acid concentrations in fetal fluids of pigs. AB - Free amino acid concentrations were determined in amniotic and allantoic fluids and in fetal and maternal uterine plasma of pigs during gestation. Glycine and glutamine were the most abundant free amino acids in maternal uterine arterial and fetal umbilical venous plasma, respectively. Plasma fetal/maternal ratios were remarkably < 1 for glycine, > 2 for glutamine and = 1 for total free alpha amino acids. Glutamine was the most abundant free amino acid in amniotic fluid during the first 3 mo of gestation. The concentrations of most amniotic fluid amino acids progressively decreased with increasing gestational age from 45 to 110 d. Glycine was the most predominant amino acid in allantoic fluid on d 30 of gestation but decreased on d 45 and 60 of gestation. This decrease in allantoic fluid glycine coincided with a marked increase in concentrations of both ornithine and arginine, which accounted for 34-36% of total free alpha-amino acids in allantoic fluid. Such an abundance of ornithine and arginine in allantoic fluid has not been reported for any other biological fluid. On d 90 and 110 of gestation, glycine again became the most abundant amino acid in allantoic fluid, or 47 and 29% of total free alpha-amino acids, respectively. Urea concentrations in amniotic and allantoic fluids progressively increased during the second half of pregnancy, indicating enhanced fetal protein metabolism. Our results demonstrate the abundance of glutamine in fetal plasma, the unusual abundance of ornithine, arginine and glycine in allantoic fluid and developmental changes in plasma fetal/maternal ratios of amino acids.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7472668 TI - Dietary maltitol increases calcium content and breaking force of femoral bone in ovariectomized rats. AB - Maltitol is a disaccharide alcohol generated by hydrogenation of maltose and exhibiting resistance to intestinal disaccharidases. We demonstrated previously that maltitol stimulates transepithelial transport of calcium in the ileum, accompanied by an elevation of intestinal calcium absorption as well as calcium retention in the body. In this study, we examined whether the maltitol-induced increase in intestinal calcium absorption leads to an alteration of the physical properties of bones in rats subjected to ovariectomy. We used this study as a simulation model for postmenopausal females who are at risk for osteoporosis. Following the intake of a low-calcium diet for 28 d ovariectomized rats were fed diets containing either 10% maltose (control) or 10% maltitol, together with increased amounts of calcium (0.3% in Experiment 1 and 1.2% in Experiment 2) for 21 d. Balance studies performed during the final 5-d (Experiment 1) or 2-d (Experiment 2) period of the experiments showed that maltitol increased intestinal calcium absorption and retention. The breaking force of femoral bones was significantly elevated (by 5-7%) in animals fed the maltitol diet compared with that in rats fed the maltose diet. The calcium content in the femoral bones as well as the mineral bone density of the tibial metaphysis was also elevated in rats fed the maltitol diet. These results indicate that maltitol stimulates the intestinal absorption of dietary calcium leading to an increase in calcium content in the bone, and coinciding with the elevation of the breaking strength of the bone in ovariectomized rats. PMID- 7472669 TI - Zinc status specifically changes preferences for carbohydrate and protein in rats selecting from separate carbohydrate-, protein-, and fat-containing diets. AB - This study examined how macronutrient intake preferences were specifically altered in the loss of appetite caused by experimentally produced zinc deficiency. Outbred female rats were allowed to freely select from simultaneously provided carbohydrate-, protein-, and fat-rich diets to provide themselves with an acceptable total diet. Rats were divided into two groups and provided the three diets containing either adequate (30 mg/kg; Zn+) or deficient (1 mg/kg; Zn ) levels of zinc (Zn). After 28 d, rats offered the Zn- diet were returned to a Zn+ diet (Zn repletion). Intakes from each of the three macronutrient diets were measured to determine macronutrient preferences of Zn-adequate, Zn-deficient, and Zn-repleted rats. In two 28-d studies involving a total of 66 rats, total metabolizable energy intake in Zn deficient rats was between 20 and 35% lower than in Zn+ rats, and carbohydrate intake accounted for essentially 100% of the lower energy intake. Fat and protein intakes were not affected by Zn deficiency. When Zn-deficient rats were repleted with Zn by providing diets containing adequate Zn, carbohydrate intake was restored to normal levels after 1 d of feeding. A transient difference in protein intake was noted during the repletion period, peaking during d 2-4 of repletion. Protein intake increased by more than 50% during this period. We hypothesize that specific changes in macronutrient intake patterns during development and recovery from Zn deficiency may be reflections, at least in part, of Zn-mediated changes in the central control of appetite. PMID- 7472670 TI - Cecal infusion of nutrients improves nutritional status of rats. AB - The role of colonic fermentation in providing energy was investigated in rats with small bowel transection (T) or 80% resection (SBR). Rats were randomized to receive for 12 d either saline (S) or the enteral solution (E) through a cecostomy to meet 30% of energy requirement; the rest (70%) was provided by parenteral nutrition. Although SBR-S rats lost weight significantly compared with d 1 of the study, SBR-E rats gained. Significantly greater carcass wet weight and fat were found in SBR-E and T-E rats compared with SBR-S and T-S rats. SBR-E and T-E rats had significantly greater colonic mucosal dry weight and protein compared with SBR-S and T-S rats. Cecal short-chain fatty acid (SCFA) contents were also significantly higher in SBR-E and T-E rats compared with SBR-S and T-S rats. There was no significant effect of surgery (T vs. SBR) on any of the variables studied. These results suggest that the products of fermentation of an enteral solution infused through a cecostomy contribute substantially to energy requirement, maintenance of body composition and nutritional status of rats. PMID- 7472671 TI - Executive summary from the report: analysis of adverse reactions to monosodium glutamate (MSG) PMID- 7472672 TI - Susceptibility for hydroperoxide formation of phosphatidylcholine and phosphatidylethanolamine in liposomes. AB - To compare the peroxidative susceptibilities of phosphatidylcholine (PC) and phosphatidylethanolamine (PE) in liposomes, multilamellar vesicles (MLVs) were prepared with equimolar L-alpha-dilinoleoyl PC (DLPC) and L-alpha-dilinoleoyl PE (DLPE), and with soya PC and soya PE having a uniform constituent fatty acids. The hydroperoxide formation at 37 degrees C in the presence of a water-soluble radical initiator was examined by chemiluminescence-high-performance liquid chromatography (CL-HPLC), and the effect of heterogeneous distribution of PC and PE on peroxidation was investigated. No difference was found between the hydroperoxidation of PC and PE in MLVs systems, except that soya PC was more susceptible to peroxidation than soya PE in the L-alpha-dipalmitoyl PC (DPPC) based liposomes. No correlation was found between the amount of phospholipids distributed in the external leaflet of MLVs and hydroperoxide formation. This result suggested that the unsaturation of constituent fatty acids in phospholipids is more important than the difference in the polar head group of phospholipids regarding their peroxidizabilities in liposomes. PMID- 7472673 TI - Effects of fructooligosaccharides on the absorption of iron, calcium and magnesium in iron-deficient anemic rats. AB - We investigated the effects of fructooligosaccharides (FO)-feeding on the absorption of iron (Fe), calcium (Ca) and magnesium (Mg) and on the biochemical parameters in Fe-deficient anemic rats. Fe-deficient anemic rats were made by feeding an Fe-deficient diet for 3 weeks. Then these Fe-deficient rats were fed an experimental diet that contained one of two levels of Fe (15 or 30 mg/kg diet), in the form of ferric pyrophosphate, and one of two levels of FO (0 or 50 g/kg diet) for 2 weeks. After the rats were fed these experimental diets, FO feeding increased the hematocrit ratio, the concentration of hemoglobin and the hemoglobin regeneration efficiency during the first week. Also, the apparent absorption of Fe was increased by FO-feeding. The levels of Fe in the diet did not affect the absorption of Ca and Mg. However, FO-feeding increased the absorption of Ca and Mg. FO-feeding lowered the pH and raised the solubility of Fe, Ca and Mg in the cecal contents, suggesting that those increasing effects of FO-feeding on absorption of these minerals is correlated with fermentation of FO in the large intestine, namely, the cecum and colon. We concluded that FO-feeding improved recovery from anemia and increased the absorption of Fe, Ca and Mg in Fe deficient anemic rats. PMID- 7472674 TI - Repression of fat-dependent intestinal apo A-IV mRNA abundance by medium chain triacylglycerols and proteins, and elevation by carbohydrates of fat-dependent apo A-IV transport in suckling rat pups. AB - Nutrients regulating intestinal apo A-IV synthesis and secretion in developing rats have not been clearly defined. We examined the effect of ingestion of fats, carbohydrates and proteins on the abundance of intestinal apo A-IV mRNA and the serum concentration of apo A-IV in 14-day-old suckling rat pups fasted overnight. In pups ingesting long-chain fatty acid-fat (soybean oil, triolein: LCT), there was a prompt elevation of the mRNA at 1.5 h after ingestion, although a graded dose of soybean oil did not result in a comparable elevation of the apo A-IV mRNA. In pups on MCT, but not trilaurin, there was a repression of the LCT dependent elevation of the message. In pups on Intralipid (composed of soybean oil, lecithin and glycerol), mRNA was elevated at 6 h after ingestion. Administration of Intralipid with lactose, glucose, fructose and sucrose induced a rapid elevation of mRNA together with elevation of serum apo A-IV, although administration of casein, whey proteins and soybean proteins resulted in repression of the Intralipid-dependent mRNA elevation. The message correlated weakly to the serum apo A-IV and triacylglycerols and with no correlation to intestinal fat accumulation. These results suggest that metabolic events following the ingestion of milk components modulate intestinal apo A-IV expression in developing rats, possibly through mucosal fatty acid utilization. PMID- 7472675 TI - Immobilization of lipases on polyethylene and application to perilla oil hydrolysis for production of alpha-linolenic acid. AB - The conditions of immobilization of lipase on the polyethylene powder were examined and the immobilized lipase was used for effective production of alpha linolenic acid from perilla oil hydrolysis. Using the polyethylene powder containing micropores in its structure enabled immobilization of lipases. Ethanol pretreatment increased the immobilization efficiency. The efficiency of immobilization of six different lipases was 67-91%. On hydrolysis of perilla oil, the activity of the immobilized lipase was nearly the same as that of free lipase. Among the enzymes used, the Candida rugosa lipase hydrolyzed perilla oil most effectively. When the immobilized C. rugosa lipase was used 10 times for perilla oil hydrolysis by the batch reaction method (1 batch, 24-h reaction at 37 degrees C), there was little decrease in its activity. These results suggest that polyethylene-immobilized lipase can be used for the effective hydrolysis of perilla oil to produce alpha-linolenic acid. PMID- 7472676 TI - n-3 and n-6 fatty acid intake and serum phospholipid fatty acid composition in middle-aged women living in rural and urban areas in Okayama Prefecture. AB - Dietary fatty acids and serum lipids were evaluated in 68 middle-aged women living in the northern, rural area of Okayama Prefecture, and were compared with the values obtained from 65 urban women from the southern part of this prefecture. A higher level in HDL cholesterol and a lower atherogenic index were observed in the rural women. The percent of energy intake as fat was lower (20.4 +/- 0.8% vs. 23.2 +/- 0.7%) and that of carbohydrate was greater in the rural group. Eicosapentaenoic (EPA, 0.41 +/- 0.04 g/day) and docosahexaenoic acid (DHA, 0.70 +/- 0.08 g/day) intakes were significantly higher in the rural subjects than in the urban group. Significantly higher DHA levels and n-3/n-6 fatty acid ratios in serum total phospholipids were found in rural women in their fifties and the sixties compared to urban women. Dietary linoleic acid (LA) amounts were positively correlated with LA (p < 0.05), and negatively with the EPA (p < 0.05) and DHA (p < 0.01) contents of serum total phospholipids. These results suggest that the traditional Japanese diet, containing little fat but enriched in complex carbohydrates and n-3 fatty acids of marine origin, may be related to the low atherogenic index in this rural area. PMID- 7472677 TI - Amino acids and glucose differentially increased extracellular 5 hydroxyindoleacetic acid in the rat brain. AB - To reveal the role of serotonergic neurons in the regulation of feeding, the levels of 5-hydroxyindoleacetic acid (5-HIAA), a metabolite of serotonin, in the striatum and the hypothalamus were continuously monitored by an in vivo microdialysis technique. Intake of 20% casein diet did not induce significant changes in the 5-HIAA level in these regions. When rats were fed on 5% casein diet (83.5% carbohydrate diet) for 2 h, the level of 5-HIAA in the striatum gradually increased and reached a maximum (226 +/- 44% of basal level, M +/- SEM, n = 7) at 4 h after stopping the diet. In the medial hypothalamus, its level also increased to 183 +/- 19% (n = 10) at 2 h after starting the diet. On the other hand, a 60% casein diet increased the level of 5-HIAA in the lateral hypothalamus to 138 +/- 19% (n = 10) at 2 h after starting the diet. The intravenous infusion of each of these nutrients, glucose, amino acid mixture or lipid, produced more rapid elevation of the 5-HIAA level than oral intake of the diets. When rats were infused with glucose, its level in the striatum continued to be elevated. In the medial hypothalamus, glucose infusion increased 5-HIAA to the maximum (189 +/- 38%, n = 7) at 4 h after starting infusion. In contrast, serotonergic neurons in the lateral hypothalamus seemed to respond only to infusion of the amino acid mixture, and the level of 5-HIAA reached 163 +/- 14% (n = 5) of the basal level at 1 h after starting the infusion. These results suggest that rapid elevation of glucose or amino acids may independently stimulate serotonin metabolism in these brain areas, participating in the feedback regulation of nutrient intake. PMID- 7472678 TI - Effect of feeding clofibrate-containing diet on the hepatic NAD+ level in rats. AB - Feeding rats with a diet containing 0.25% clofibrate for 2 weeks elevated the hepatic NAD+ and total nicotinate levels significantly. Other peroxisome proliferators, such as 2-(4-chlorophenoxy)propionic acid and di(2 ethylhexyl)phthalate, had similar effects. When rats were fed the control diet without clofibrate for 1 week after 2 weeks of the clofibrate diet, the hepatic NAD+ level returned to the control value. Muscular NAD+ content was not affected by the peroxisome proliferators. The results were discussed in relation to induction of peroxisomal beta-oxidation enzymes by the peroxisome proliferators. PMID- 7472679 TI - Selenium levels and glutathione peroxidase activities in blood in an andean high altitude population. AB - Selenium (Se) levels in blood (whole blood, erythrocytes and serum) and blood glutathione peroxidase (GSH-Px) activity were investigated in people living at high altitude in Bolivia (4,000 m above sea level). These parameters were compared to those of people living at low altitude (300 m above sea level). The Se levels in whole blood of the high-altitude subjects did not differ significantly from those of the low-altitude subjects. However, the Se levels in erythrocytes were significantly lower in the high-altitude subjects than in the low-altitude subjects, whereas serum Se levels were higher in the high-altitude subjects than in the low-altitude subjects. GSH-Px activity (Unit/g Hb) was significantly lower in the high-altitude subjects than in the low-altitude subjects. The mean corpuscular Hb concentration (MCHC), an indicator of the age of erythrocytes, in the high-altitude subjects was significantly higher than in the low-altitude subjects. These results show that the GSH-Px activity in the blood of the high-altitude subjects is relatively low. This may be due to their aged erythrocytes and/or to relatively low Se intake in the high-altitude population compared with low-altitude population. PMID- 7472680 TI - Pyridoxal 5'-phosphate modulates expression of cytosolic aspartate aminotransferase gene by inactivation of glucocorticoid receptor. AB - The level of mRNA for cytosolic aspartate aminotransferase (cAST) in the liver of vitamin B6-deficient rats was found to be 7-fold higher than that of the control rats. The administration of hydrocortisone to adrenalectomized vitamin B6 deficient rats induced expression of hepatic cAST mRNA and the induction was suppressed by the simultaneous administration of pyridoxine. Since the 5' regulatory region of the rat cAST gene contains several sequences showing homology to glucocorticoid-responsive elements, we synthesized an oligonucleotide probe of glucocorticoid-responsive element sequence and assayed the binding activity of liver nuclear extract to the oligonucleotide by gel mobility shift analysis. We found that the binding activity of nuclear extract prepared from the liver of vitamin B6-deficient rats was far greater than that of the control rats, indicating that the DNA-binding activity of glucocorticoid receptor was enhanced by vitamin B6 deficiency. We further found that preincubation of the nuclear extract from the vitamin-deficient liver with pyridoxal 5'-phosphate brought about a rapid and extensive decrease in the binding of the extract to the glucocorticoid-responsive element. Congeners of pyridoxal phosphate, such as pyridoxamine 5'-phosphate, pyridoxal, pyridoxamine and pyridoxine, did not show an inhibitory effect. These observations suggest that pyridoxal 5'-phosphate modulates cAST gene expression by inactivating the binding activity of glucocorticoid receptor to glucocorticoid-responsive elements. PMID- 7472681 TI - Effects of chemical sympathectomy on lipoprotein lipase activities in peripheral tissues of rats fed high fat diets consisting of different fats. AB - We have previously reported that the feeding of a beef tallow diet gives lower lipoprotein lipase activities in the interscapuler brown adipose tissue, heart and soleus muscle compared with feeding of a safflower oil diet. We have also suggested that sympathetic activities (norepinephrine turnover rates and beta adrenergic receptor bindings) in these peripheral tissues were lower in the beef tallow diet group. To confirm the effects of dietary fats on the lipoprotein lipase activities in these tissues related to sympathetic activities, rats were chemically sympathectomized by treatment with 6-hydroxydopamine. The sympathectomized rats and control rats were meal-fed with a beef tallow diet or a safflower oil diet for 8 weeks. Sympathectomy abolished the differences in lipoprotein lipase activities in the interscapuler brown adipose tissue, heart and soleus muscle between two dietary groups. The percentage of body fat was increased by sympathectomy, resulting in no difference between the beef tallow diet group and the safflower oil diet group. These results suggest that intake of the beef tallow diet decreased lipoprotein lipase activities by reducing the sympathetic activities. PMID- 7472682 TI - Visible light-sensitized oxidation of arachidonic acid in the presence of inperatorin. AB - To understand the mechanism of the phototoxic effects of inperatorin, a psoralen derivative used as a pigmentation agent, we have investigated the photosensitized oxidation of arachidonic acid (ARA) by irradiation with visible light (> 400 nm) in the presence of inperatorin. HPLC and GC/MS analyses of the products showed the formation of many hydroperoxyeicosatetraenoic acids (HPETEs) including the products of lipoxygenase-catalyzed reactions such as 5- and 15-HPETEs, which are the precursors of chemical mediators such as leukotrienes and lipoxins, during the reaction. Active oxygen scavening agents such as D-mannitol, superoxide dismutase, and beta-carotene inhibited the formation of the oxidation products, indicating that the oxidation reaction was mediated by various active oxygen species. These results suggest that the phototoxic effects of inperatorin could also be induced by visible light and could be explained at least partially in terms of inflammation initiated by the biologically active HPETEs arising from photosensitized oxygenation reactions of ARA with the drug. PMID- 7472683 TI - Comparison of a disease-specific and a generic severity of illness measure for patients with community-acquired pneumonia. AB - OBJECTIVE: To compare the performances of a disease-specific severity of illness index and a prototypical generic severity of illness measure, MedisGroups Admission Severity Groups (ASGs), for patients with community-acquired pneumonia. DESIGN: A retrospective database study. PATIENTS: Adult patients (aged > or = 18 years) with an ICD-9-CM principal diagnosis of pneumonia in 78 MedisGroups Comparative Database hospitals. METHODS: The pneumonia severity of illness index (PSI) was developed to predict hospital mortality using logistic regression analyses in a 70% random sample of study patients. The performances of the PSI and the generic severity measure were assessed among the remaining 30% of patients by comparing observed mortalities within the five PSI and ASG severity classes, and areas under their receiver operating characteristic (ROC) curves. Both the PSI and the generic severity measure were used to estimate the 95% confidence interval of the expected number of deaths in each of the 78 study hospitals. Hospitals with an observed number of deaths outside these limits were identified as outliers. RESULTS: There were 14,199 study patients who had community-acquired pneumonia, and 1,542 (10.9%) died during hospitalization. In comparison with the generic severity measure, the PSI more accurately identified patients at extremely low risk of death, and had a larger area under its ROC curve (0.84 vs 0.79; p < 0.0001). Of the 78 study hospitals, 17 (21.8%) were classified as outliers for mortality by at least one severity adjustment system. Among the 11 low-outlier hospitals, six were classified by the generic severity measure alone, two by the PSI alone, and three by both systems; among the six high-outlier hospitals, one was classified by the generic measure alone, three by the PSI alone, and two by both systems. CONCLUSIONS: The PSI provided more accurate estimates of hospital mortality and classified different hospital outliers for mortality than did the generic severity of illness measure for patients with community-acquired pneumonia. PMID- 7472684 TI - "Strong medicine": Cambodian views of medicine and medical compliance. AB - OBJECTIVE: To identify important determinants of noncompliance among Cambodian refugees. DESIGN: Open-ended interviews. SETTING: University-affiliated ambulatory care clinic in an inner-city hospital. PARTICIPANTS: Thirty adult Cambodian refugees (15 men and 15 women) who were regular utilizers of a refugee clinic. MEASUREMENTS AND MAIN RESULTS: Sixty-seven percent of the participants described being noncompliant with at least one of their medications. Four common causes of noncompliance were identified: 1) misunderstanding the intent of the medication; 2) side effects; 3) concern about the effect of medication on "internal strength"; and 4) Cambodian ideas about pharmacokinetics. CONCLUSIONS: Noncompliance was commonly reported by the Cambodian respondents. Noncompliance was both intentional and unintentional; the unintended noncompliance derived from patients' attempting to comply with therapy according to Cambodian ideas about the body and Western medication. This article suggests clinical approaches to enhance compliance with prescribed regimens among recent Cambodian immigrants and refugees. PMID- 7472685 TI - Physicians' psychosocial beliefs correlate with their patient communication skills. AB - OBJECTIVE: To assess the relationship between physicians' beliefs about the psychosocial aspects of patient care and their routine communication with patients. PARTICIPANTS AND SETTING: Fifty community primary care physicians participating in a continuing medical education program and 473 of their patients in Portland, Oregon. METHODS: Routine office visits were audiotaped and analyzed for communication behaviors and emotional tone using the Roter Interactional Analysis System (RIAS). Physician beliefs about psychosocial aspects of care were measured using a self-report questionnaire with a five-point Likert scale. Attitudes were correlated with communication behaviors using the Pearson correlation coefficient. RESULTS: Physicians' attitudes toward psychosocial aspects of care were associated with both physician and patient dialogue in visits. The physicians who had positive attitudes used more statements of emotion (i.e., empathy, reassurance) (p < 0.05) and fewer closed-ended questions (p < 0.01) than did their colleagues who had less positive attitudes. The patients of the physicians who had positive attitudes more actively participated in care (i.e., expressing opinions, asking questions), and these physicians provided relatively more psychosocial and less biomedical information (p < 0.05). CONCLUSION: Physician beliefs about psychosocial aspects of patient care are associated with their communication with patients in routine office visits. Patients of physicians with more positive attitudes have more psychosocial discussions in visits than do patients of physicians with less positive attitudes. They also appear more involved as partners in their care. These findings have implications for medical educators, teachers, and practicing physicians. PMID- 7472686 TI - Predicting cutaneous hypersensitivity reactions to cotrimoxazole in HIV-infected individuals receiving primary Pneumocystis carinii pneumonia prophylaxis. AB - OBJECTIVES: To measure the incidence of cutaneous hypersensitivity reactions to cotrimoxazole in the setting of primary Pneumocystis carinii pneumonia (PCP) prophylaxis: to measure the incidence of severe reactions: and to identify predictors for these outcomes. DESIGN: Retrospective cohort study. SETTING: One university-based outpatient HIV clinic and one university-affiliated internal medicine and infectious disease medical practice. PATIENTS: Two hundred thirty six HIV-infected individuals receiving cotrimoxazole for primary PCP prophylaxis. INTERVENTIONS: None. MAIN OUTCOME MEASURE: Occurrence of a cutaneous hypersensitivity reaction, defined as rash, fever, or pruritus that resulted in permanent discontinuation of cotrimoxazole. Severe reactions were defined as those resulting in hospital admission or systemic treatment with a corticosteroid. Cox regression was used to calculate relative rates (RRs) and 95% confidence intervals (CIs) for a number of clinical and laboratory variables. MEASUREMENTS AND MAIN RESULTS: Forty-eight (20%) subjects developed cutaneous hypersensitivity reactions, with six (12.5%) of these being severe. In the unadjusted analysis, the following factors demonstrated at least borderline association: male gender [RR (95% CI) = 0.46 (0.21-0.99)], higher CD4 percentage [RR (95% CI) = 0.95 (0.90-1.00)], syphilis history [RR (95% CI) = 0.37 (0.13 1.04)], and higher total protein [RR (95% CI) = 0.70 (0.45-1.09)]. Adjustment for potential confounding by measured variables did not meaningfully change these results. CONCLUSIONS: Cutaneous hypersensitivity reactions to cotrimoxazole in the setting of primary PCP prophylaxis are common. Although male gender, higher CD4 percentage, syphilis history, and higher total protein have at least borderline associations with these reactions, routinely collected clinical and laboratory variables do not appear to be sufficiently associated with the reactions to permit development of a clinically useful prediction rule. PMID- 7472687 TI - A comparison of the methods and criteria used by traditional and primary care internal medicine programs to select residents. AB - OBJECTIVE: To determine whether there are differences in the methods and criteria used by primary care and traditional internal medicine programs to select first year residents. DESIGN: A questionnaire was sent to primary care and traditional internal medicine program directors, who were asked to rank in importance ten documents of an applicant's file and to score the relative importance, on a scale of -5 to +5, of 21 candidate traits of four types: academic, demographic, personal, and career goal. SETTING: Programs at institutions (n = 54) that have categorical residency programs in both traditional and primary care internal medicine. PARTICIPANTS: Of 108 questionnaires, the overall response rate was 81%, with 40 pairs (74%) of matched respondents. Seventy-two percent of the responding institutions were university-administered. RESULTS: Primary care and traditional programs use similar methods to process applicants, rank similarly ten documents in an applicant's file, and value academic success during the clinical years as the most important candidate trait. Compared with traditional tracks, primary care tracks place greater emphasis on a candidate's career goals and select for candidates planning to pursue primary care careers (3.9 +/- 1.4 vs 0.9 +/- 1.5, p < 0.001), enter practice (1.4 +/- 1.5 vs 0.1 +/- 1.2, p < 0.001), or serve medically indigent populations (2.7 +/- 1.5 vs 1.2 +/- 1.2, p < 0.001). Primary care programs rate negatively candidates who intend to subspecialize, whereas traditional programs view them almost neutrally (-1.8 +/- 2.2 vs 0.5 +/- 1.5, p < 0.001). CONCLUSION: Primary care and traditional track internal medicine programs use similar methods to select residents and both rank academic achievement during the clinical years as a candidate's most important attribute. However, only primary care programs strongly select for candidates on the basis of their career plans and in particular prefer candidates who are committed to pursuing primary care careers and serving the medically indigent. PMID- 7472688 TI - Drug companies and continuing medical education. PMID- 7472690 TI - The night float system of resident on call: what do the nurses think? AB - In 1989, the internal medicine residency training program at St. John's Episcopal Hospital changed from a standard one-in-four on-call system to a night float system of resident on call. Using a confidential questionnaire that assessed the opinions of medical nurses, the nurses were asked which system they preferred, and how the change to a night float system of resident on call affected medical resident performance. A significant majority of the nurses preferred night float over a standard system of resident on call. Most nurses responded that because night float allows residents to get more rest, residents make fewer mistakes and are easier to work with. Residents on night float were not considered to be more knowledgeable. The responders indicated that night float did not create confusion about which resident to call for a patient problem. PMID- 7472689 TI - Ability of primary care physicians to diagnose and manage Pneumocystis carinii pneumonia. AB - This study assesses the ability of primary care physicians to diagnose and manage Pneumocystis carinii pneumonia (PCP) in a standardized patient (SP) with unidentified HIV infection. One hundred thirty-four primary care physicians from five Northwest states saw an SP with unidentified HIV infection who presented with symptoms, chest radiograph, and arterial blood gas results classic for PCP. Seventy-seven percent of the physicians included PCP in their differential diagnoses and 71% identified the SP's HIV risk. However, only a minority of the physicians indicated that they would initiate an appropriate diagnostic evaluation or appropriate therapy: 47% ordered a diagnostic test for PCP, 31% initiated an antibiotic appropriate for PCP, and 12% initiated an adequate dose of trimethoprim - sulfamethoxazole. Only 6% of the physicians initiated adjunctive prednisone therapy, even though prednisone was indicated because of the blood gas result. These findings suggest significant delay in diagnosis and treatment had these physicians been treating an actual patient with PCP. PMID- 7472691 TI - Who leaves against medical advice? AB - Patients who leave hospitals against medical advice (AMA) frustrate physicians and may put themselves at medical risk. A case-control study was conducted to characterize the factors associated with AMA discharges from an impatient medical service. Logistic regression analysis indicated that not having a primary care physician and previous AMA discharge were significantly associated with leaving AMA. The patients most often stated that they were leaving because they "felt better" or had personal or financial obligations. However, the patients leaving AMA were more likely to return for care within the next week than were the control patients. PMID- 7472692 TI - Examining the validity of severity measures in today's health policy context. PMID- 7472693 TI - Different questions beg different methods. PMID- 7472694 TI - How important is context in teaching interviewing? PMID- 7472695 TI - Use of pneumococcal vaccine. PMID- 7472696 TI - Things that ring in the night. PMID- 7472697 TI - Benefits of computer diagnostic assistants. PMID- 7472698 TI - Comparison of a lifestyle modification program with propranolol use in the management of diastolic hypertension. AB - OBJECTIVE: To compare the management of mild diastolic hypertension (90 to 104 mm Hg) using a nonpharmacologic intervention with that using propranolol or placebo. DESIGN: Randomized, placebo-controlled trial with a 2 x 2 factorial design. SETTING: University-based ambulatory care center. PARTICIPANTS: Two hundred seven men and 105 women, 22 to 59 years of age, 73% white, who had mild diastolic hypertension untreated for at least eight weeks. INTERVENTIONS: 1) a multicomponent lifestyle modification intervention (lifestyle focus group, or LFG) administered in eight weekly meetings + placebo, 2) LFG + propranolol, 3) propranolol alone, and 4) placebo alone, followed for 12 months. MEASUREMENTS: Systolic blood pressure (SBP), diastolic blood pressure (DBP), and self-reported adverse effects at each of nine follow-up visits; fasting total cholesterol, triglycerides, and glucose at baseline and 12 months; 24-hour urine sodium (Na+) and potassium (K+), three-day food records and physical activity questionnaire at three and 12 months; and a quality of life questionnaire at 12 months. MAIN RESULTS: The mean decreases in DBP at 12 months were: 8.5 mm Hg in the LFG + propranolol group; 7.7 mm Hg in the propranolol-only group; 5.9 mm Hg in the placebo-only group; and 5.4 mm Hg in the LFG + placebo group. Repeated-measures analysis of covariance showed that level of baseline DBP (p < 0.0001), time of follow-up (p < 0.0001), and propranolol use (p < 0.0001) were significantly associated with a decrease in DBP at 12 months. Despite reductions in urinary Na+ (-35 mEq; 95% CI = -50, -19), dietary Na+ (-521 mg; 95% CI = -710, -332), total calories ingested (-238; 95% CI = -335, -140), and weight (-1.4 lb; 95% CI = 3.7, +0.8), and significant increases in dietary K+ (+294 mg; 95% CI = +107, +480) and in mets-minutes of exercise (+43; 95% CI = +20, +67) at three months, assignment to the LFG intervention had no effect on DBP at three or 12 months. The subjects assigned to take propranolol more frequently reported fatigue during ordinary activities, sleep disturbance, decrease in sexual activity, and depressed feelings, when compared with the subjects taking placebo, but the numbers of study withdrawals did not differ by drug assignment. No significant difference in total cholesterol and glucose levels was observed by group assignment. Triglycerides increased significantly in the subjects assigned to propranolol (mean difference = +20 mg/dL; 95% CI of difference +1.5, +39). There was no difference in the responses to 21 quality of life items between the subjects assigned to propranolol and those assigned to placebo. CONCLUSIONS: This multicomponent lifestyle modification intervention was unable to promote persistent behavior changes and thus was inferior to propranolol therapy for the treatment for mild diastolic hypertension. Future research should focus on single modifiable factors to lower blood pressure. PMID- 7472699 TI - Expert ratings of primary care goals and objectives. AB - OBJECTIVE: To develop consensus on proficiencies internal medicine residents should master in the area of primary and managed care. DESIGN: A draft compendium of primary care educational objectives including important clinical topics was developed at the Sepulveda Veterans Health Administration Medical Center Pilot Ambulatory Care and Education (PACE) Program as part of a local and regional primary care curricular review. Fifty-one experts, including leaders in the Society of General Internal Medicine, the Association of Program Directors in Internal Medicine, the American College of Physicians, general internal medicine division chiefs, and Veterans Affairs (VA) associate chiefs of staff for ambulatory care rated the compendium. MEASUREMENTS AND MAIN RESULTS: Eleven objectives and nine clinical topics were rated "critically important" (4.7 or above on a five-point scale). General internal medicine chiefs and associate chiefs of staff for ambulatory care judged them to be covered adequately in fewer than half of the 17 VA Western Region-affiliated internal medicine programs. Forty-five objectives and 77 clinical topics were considered at least somewhat important to the education of general internal medicine residents in primary care. The VA raters reported that in the prior academic year, their housestaffs had spent between 21% (postgraduate year I) and 33% (postgraduate year III) of their time in ambulatory care settings. CONCLUSION: With the emphasis on primary and managed care, there is a need for national consensus on educational objectives in primary care general internal medicine. This review provides educators with a benchmark to test the adequacy of their institutions' curricula in primary care internal medicine. PMID- 7472700 TI - How do medical residents discuss resuscitation with patients? AB - OBJECTIVE: To describe how medical residents discuss do-not-resuscitate (DNR) orders with patients. DESIGN: Prospective observational study. SETTING: Inpatient medical wards of one university tertiary care center, one urban city public hospital, and one Veterans Affairs medical center. PARTICIPANTS: Thirty-one medical residents self-selected 31 of their English-speaking, competent patients, with whom they had DNR discussions. MEASUREMENTS: Three independent observers rated audiotaped discussions about DNR orders between the medical residents and their patients. Ratings assessed whether the physicians met standard criteria for requesting informed consent (e.g., disclosed the nature, benefits, risks, and outcomes), addressed the patients' values, and attended to the patients' emotional concerns. MAIN RESULTS: The physicians often did not provide essential information about cardiopulmonary resuscitation (CPR). While all the physicians mentioned mechanical ventilation, only 55% mentioned chest compressions and 32% mentioned intensive care. Only 13% of the physicians mentioned the patient's likelihood of survival after CPR, and no physician used a numerical estimate. The discussions lasted a median of 10 minutes and were dominated in speaking time by the physicians. The physicians initiated discussions about the patients' personal values and goals of care in 10% of the cases, and missed opportunities to do so. CONCLUSIONS: Medical ethicists, professional societies, and the public recommend more frequent discussions about DNR orders. Even when housestaff discuss resuscitation with patients, they may not be accomplishing the goal of increasing patient autonomy. Research and education must focus on improving the quality, as well as the quantity, of these discussions. PMID- 7472701 TI - Policies, practices, and attitudes of North American medical journal editors. AB - OBJECTIVE: To describe U.S. and Canadian medical journals, their editors, and policies that affect the dissemination of medical information. DESIGN: Mailed survey. PARTICIPANTS: Senior editors of all 269 leading medical journals published at least quarterly in the United States and Canada, of whom 221 (82%) responded. MAIN MEASURES: The questionnaire asked about characteristics of journal editors and their journals and about journals' policies toward peer review, conflicts of interest, prepublication discussions with the press, and pharmaceutical advertisements. RESULTS: The editors were overwhelmingly men (96%), middle-aged (mean age 61 years), and trained as physicians (82%). Although 98% claimed that their journals were "peer-reviewed," the editors differed in how they defined a "peer" and in the number of peers they deemed optimal for review. Sixty-three percent thought journals should check on reviewers' potential conflicts of interest, but only a minority supported masking authors' names and affiliations (46%), checking reviewers' financial conflicts of interest (40%), or revealing reviewers' names to authors (8%). The respondents advocated discussion of scientific findings with the press (84%), but only in accord with the Ingelfinger rule, i.e., after publication of the article (77%). Fifty-seven percent of the editors agreed that journals have a responsibility to ensure the truthfulness of pharmaceutical advertisements, and 40% favored subjecting advertisements to the same rigorous peer review as scientific articles. CONCLUSIONS: The responding editors were relatively homogeneous demographically and professionally, and they tended to support the editorial status quo. There was little sentiment in favor of tampering with the current peer-review system (however defined) or the Ingelfinger rule, but a surprisingly large percentage of the respondents favored more stringent review of drug advertisements. PMID- 7472702 TI - Women's health and enhancing generalism in internal medicine. PMID- 7472703 TI - Accuracy and reliability of apical S3 gallop detection. AB - This study assessed physician performance in detecting the apical S3 gallop using a cardiology patient simulator. Six physicians (two cardiology fellows, two medicine residents, and two attending physicians) performed two sets of 24 cardiac examinations that included the presence or absence of an apical S3 gallop. All the examiners were able to significantly alter the prior odds of an apical S3 gallop's being present, but the cardiology fellows had higher sensitivities. Sensitivity was lower for detecting soft S3 gallops, and specificity was lower when a diastolic murmur was also present. Physician performance in detecting apical S3 gallops is variable, but can be excellent. PMID- 7472704 TI - Alcohol consumption among HIV-infected patients. AB - This prospective, cohort study analyzed the prevalence of alcoholism and patterns of alcohol intake over time in a cohort of HIV-infected patients, predominantly homosexual/bisexual men. One hundred eleven HIV-positive subjects were recruited from a comprehensive HIV clinic associated with a large Midwestern university hospital. Each participant completed the Michigan Alcoholism Screening Test (MAST) survey and a standardized quantity-frequency questionnaire on alcohol intake at enrollment. The quantity-frequency scale was repeated every six months for a total of 30 months. Forty-five of the 111 subjects (41%) met the criteria for alcoholism, as defined by a MAST score 5 or higher. There was a significant decrease in alcohol consumption over time, from 6.4 drinks/week in the initial time period to 3.9 drinks/week by the final time period (p < 0.001). PMID- 7472705 TI - Health insurance and utilization in Pneumocystis carinii pneumonia. AB - To examine the relationship between health insurance status and health services utilization among HIV-infected patients with active Pneumocystis carinii pneumonia (PCP), the discharge and outpatient databases of a large California medical facility were reviewed. Four hundred fifty records met the inclusion criteria. Logistic regression indicated that those who had publicly funded insurance were about half as likely (95% CI 0.28, 0.99) as were those who had private insurance to undergo bronchoscopy with bronchoalveolar lavage at the time of their first PCP episodes. Further research is required to identify the reasons for this difference. PMID- 7472707 TI - Disease management needs new paradigms. PMID- 7472706 TI - Cholesterol in patients with coronary heart disease: how low should we go? AB - CLINICAL PROBLEM: To examine the evidence supporting the recent National Cholesterol Education Program (NCEP) recommendation that low to moderate levels of cholesterol should be aggressively managed in patients with coronary heart disease (CHD). METHODS: Cohort studies and clinical trials with angiographic or clinical endpoints, that included CHD patients with low to moderate levels of cholesterol, were systematically identified through a MEDLINE search and critically reviewed. SYNOPSIS: None of the cohort studies show that a moderate level of cholesterol confers significantly increased risk of CHD death, although a pooled relative risk of 1.14 (95% CI 1.08 to 1.4) suggests that there may be a slight excess risk. Of five angiographic trials of CHD patients with moderate levels of cholesterol, two demonstrated no improvement in angiographic endpoints with intensive lipid-lowering therapy and the other three are difficult to interpret since they included other interventions in addition to the cholesterol lowering regimen. No large clinical trial with clinical endpoints has been reported for CHD patients with low to moderate levels of cholesterol. RECOMMENDATIONS: The recommendation to treat CHD patients who have low to moderate levels of cholesterol with diet or drugs is not based on convincing evidence of efficacy. This is in clear contrast to the recommendation for CHD patients with high levels of cholesterol, for whom there is definitive clinical trial evidence of benefit from cholesterol-lowering therapy. While we await clinical trial results for CHD patients with low to moderate levels of cholesterol, clinicians and patients must consider the possible disadvantages of therapy in relation to the uncertain benefit. PMID- 7472708 TI - Guardians of the lamp. PMID- 7472709 TI - How many Ryan Lukes? PMID- 7472710 TI - Marketing one, two, three. PMID- 7472711 TI - Why the interest in women's health? PMID- 7472713 TI - Insurers pursuing Medi-Cal patients. PMID- 7472712 TI - A rare breed. PMID- 7472714 TI - Ripe apple. PMID- 7472715 TI - The changing spectrum of pulmonary infections due to nontuberculous mycobacteria. AB - Although nontuberculous mycobacteria (NTM) infections are not common, there is evidence that they are becoming increasingly recognized and in some parts of the country occur more frequently than M. tuberculosis. The classic description of NTM pulmonary infection is that of a male patient with cavitary infiltrates and predisposing lung disease. With an increase in the number of reported cases of infection, there have been other syndromes described, particularly in elderly female patients without any predisposing conditions, those with asymptomatic pulmonary nodules, esophageal disease causing recurrent aspiration pneumonitis, and disseminated infection in AIDS. Therapy of M. avium complex (MAC) disease is far from definitive whereas infections due to M. kansasii can be effectively treated even in the presence of profound immunosuppression. Recognition of these infections and their recent change in epidemiology hopefully will encourage practitioners to enroll their patients into clinical protocols that will help to establish the optimal regimen(s). PMID- 7472716 TI - Benign aging or Alzheimer disease? AB - We show that in benign aging, normally functioning elders have minor neurobehavioral deficits in activities of daily living, and in their neurologic, motor and sensory status; hearing is peripherally and centrally impaired. Also, depression appears in 25%. Gentle physical exercise improves mobility, prevents falls, diminishes pain, wards off depression, reduces mortality, and increases cerebral blood flow and cognition. Diagnosis of Alzheimer disease consists of (1) proof of dementia, (2) meeting established clinical criteria, and (3) staging of severity. We describe dementia, giving tables to identify it and distinguish it from depression, and cite some reversible dementias. We report the accepted clinical inclusion criteria and exclusion criteria for this disease. We show, also with tables, the staging of severity of both dementia and Alzheimer disease as mild, moderate, or severe. PMID- 7472717 TI - Acute necrotizing fasciitis in childhood. AB - Acute necrotizing fasciitis is a rare but frequently catastrophic infection of the subcutaneous tissues and fascial sheaths. It is rare in infants and children. The case of a 3 1/2-year-old boy with acute necrotizing fasciitis caused by group A beta hemolytic streptococcus is described. PMID- 7472718 TI - AMA statement to the House Ways and Means Committee Subcommittee on Health. PMID- 7472719 TI - Coming soon: certified workplace medical plans. PMID- 7472720 TI - Is the practice of medicine getting harder? Or am I just getting older? PMID- 7472721 TI - The use of interferon for the treatment of viral hepatitis in pediatric liver transplant recipients. PMID- 7472722 TI - Drooby writes to Capitol Hill contingent. PMID- 7472723 TI - Surgical and prosthodontic training of general practitioners for single tooth implants: a study of treatments performed at four general practitioners' offices and at a specialist clinic after 2 years. AB - Both the surgical and the prosthodontic procedures will be performed by one and the same general practitioner (GP) for many implant patients, as access to specialists is limited or non-existent in a large number of places. However, treatment by GPs has not been documented to the same extent as treatment performed by specialists. This lack of knowledge was the reason for the present study, in which four GPs performed both the surgery and the prosthodontics on patients requiring single tooth implant replacements. The treatments were performed after an initial training of the GP and his/her dental assistant for 8 days. The treatments by the GPs exhibited clinical results corresponding to those achieved at a specialist clinic. The outcome indicates that complete single tooth implant treatments might be performed by GPs who have passed an initial training and demonstrated an interest in a close co-operation with specialists in oral surgery/periodontics and prosthodontics. Such a co-operation should be based on the distribution of complicated cases to the specialists and of non-complicated cases to the GPs. PMID- 7472724 TI - Self-assessment of pain and discomfort in patients with temporomandibular disorders: a comparison of five different scales with respect to their precision and sensitivity as well as their capacity to register memory of pain and discomfort. AB - Five different scales of self-assessment of pain were tested in patients with temporomandibular disorders. The precision and sensitivity and the capacity to register memory of pain and discomfort were compared for each of the five scales. The behaviour rating scale was found to be superior to the other four scales in respect of precision and sensitivity to pain and discomfort and when recording the memory of these two variables. This scale was also considered by the patients to be the most relevant and the simplest to understand. From these results, the behaviour rating scale can be recommended when measuring pain and discomfort in patients with temporomandibular disorders. PMID- 7472725 TI - Three-dimensional distortion of gold alloy castings and welded titanium frameworks. Measurements of the precision of fit between completed implant prostheses and the master casts in routine edentulous situations. AB - Thirty routine patients, provided with fixed prostheses supported by osseointegrated Branemark implants in edentulous lower jaws, were arranged into three different groups with regard to design of the metal framework. Ten patients received cast gold alloy frames and the other two groups were provided with two different designs of welded titanium frames. The fit of the completed prostheses was measured in three dimensions (3-D) in relation to the master cast, by means of a photogrammetric technique, prior to insertion. Mean 3-D distortion of the centre point of the gold cylinder was 42 (s.d. 8) microns for the cast framework. The corresponding mean distortion for the two designs of titanium frameworks was 43 (s.d. 16) and 36 (s.d. 10) microns, respectively. Least distortion was observed in vertical direction for all three designs. None of the different designs of metal frames showed a significantly better fit (P > 0.05), but the cast and oldest titanium framework design presented a much wider range of distortion. This indicated a higher risk of sectioning and resoldering during the fabrication of the prostheses as compared to the more consistently fabricated prostheses, with a new titanium framework design. PMID- 7472726 TI - The face height in adult dentate humans. A discussion of physiological and prosthodontic principles illustrated through a case report. AB - The adaptability of the stomatognathic system to alterations in the vertical dimension is discussed based on scientific reports and clinical experience. A case is reported demonstrating the stability of a therapeutically induced increase in the occlusal vertical dimension over a 10 year period. In conclusion, it is stated that moderate alterations in the occlusal vertical dimension of patients are generally well tolerated if necessary for therapeutic measures. PMID- 7472727 TI - Ceramic inlay systems: some clinical aspects. AB - Due to their inertness and unsurpassed aesthetic properties, ceramic materials are increasingly used as inlay materials. Based on a questionnaire, the frequency and character of complications related to ceramic inlay therapy were assessed and some important pitfalls and clinical relevant factors discussed. A majority of the dentists performed between 1 and 10 ceramic inlays per month using several different ceramic materials. As many as 85% of the dentists had observed complications in connection with ceramic inlays. Most frequency reported was hypersensitivity of a transient nature (15%). The material revealed an overall inlay fracture frequency of 5%, but the individual variations were high. Tooth fracture, loss of retention and other complications such as secondary caries, endodontic envolvement, colour mismatch and a surface and edge deterioration occurred with a varying but lower frequency. Failure analysis revealed a large number of causative factors among which cavity and inlay design, bonding procedures, selection of luting material and proper case selection were regarded as the most important. Due consideration to the strength of remaining tooth substance was found to be important in reducing the problem of hypersensitivity and the risk of tooth and inlay fracture. PMID- 7472728 TI - Odds ratio in the estimation of the significance of occlusal factors in craniomandibular disorders. AB - The usage of odds ratios for estimating the significance of occlusion is based on the assumption that the aetiological factors are additive and interchangeable in several different composite causes. If the factor studied is a necessary causal factor, odds ratios cannot be used in risk estimation. The present study analysed odds ratios in a sample of children half of whom had undergone an annual prophylactic elimination of occlusal interferences. The sample consisted of 178 children followed up for 5 years from the baseline age of 5 or 10. The distribution of interferences was computed for children with tenderness of the masticatory muscles ('cases'), and for children without ('controls'). Although the increase in the number of children free from interferences was not remarkable, the overall result was statistically significant (P < 0.01). None of the children free from interferences showed muscular tenderness at any of the annual examinations. Therefore, odds ratios could not be calculated in our study sample. It is concluded that estimation by means of odds ratios of the significance of occlusal factors in CMD may be misleading. PMID- 7472729 TI - Surface and colloid chemical aspects of saliva particle interactions. AB - Sections of flash-frozen saliva particle mixture were examined by transmission electron microscopy. The saliva samples used consisted of 40% by volume parotid and 60% by volume submandibular sublingual fractions. One mL of the saliva was mixed with 0.1 mL of either hydrophilic or hydrophobic particles of 0.9 microns diameter. The micrographs of the saliva and hydrophilic particles mixes showed the multiple spherical particles in the salivary network structures. Micrographs of the saliva and hydrophilic particle mixes showed the presence of few such objects; when observed, the particles were located only in the electron dense part of the salivary structures. PMID- 7472730 TI - Functional evaluation after TMJ surgery. AB - Oral function was evaluated in 30 patients (25 women and five men) after TMJ surgery by means of bite force measurements. Twenty-seven discectomies and three condylectomies had been performed. The follow-up period was, on average, 2.5 years. The mean bite force values on the function level 'biting as when chewing' was for the whole group 125.3 N on the right side and 128.8 N on the left side. The maximal bite force was on average 362.5 N and maximal endurance time on average 32 s. All test results showed big ranges. No significant differences between right and left side or operated and non-operated side could be found on the tested function level 'biting as when chewing'. The found bite force values were high in comparison with another similar study. The results were also compared with another group of patients (n = 6) who were examined before and 6 months after surgery. As far as bite force reflects oral function the results are indeed very encouraging. PMID- 7472731 TI - A 25 year longitudinal study of patients treated with removable partial dentures. AB - A 25 year longitudinal study was carried out on a number of patients fitted with removable partial dentures (RPDs) in 1969. Before the prosthetic treatment all patients were given oral hygiene motivation and instructions in order to create a high level of co-operation. The RPDs, most of which were lower bilateral distal extension dentures, were carefully planned and designed. During the first 10 years the patients were examined in our clinic at yearly intervals at which time encouragement and reinstruction regarding oral hygiene were given and various treatment procedures were performed as required. After the initial 10 years the patients were advised, for practical reasons, to continue to have yearly controls on their own initiative. Of the initial 30 patients from 1969, 23 were still alive in 1994, all of whom were examined. In five of those 23 the original RPD situation had changed more or less due mainly to general illness contracted and/or change to other therapies. In the remaining 18 patients wearing in total 20 RPDs, 13 (65%) of the original RPDs were still functioning. Seven RPDs were new with principally the same design as the original ones. Among these 18 patients the number of lost teeth, the number of new DF surfaces and the increased number of endodontically treated teeth were few. No apparent changes took place regarding the periodontal condition during the follow-up period.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7472732 TI - Occlusal interferences in retruded contact position and temporomandibular joint sounds. AB - In 193 non-patient adolescents, unilateral contacts in retruded contact position (RCP) were seen more often in girls than in boys (P < 0.001) and were more frequent in subjects with than without general joint instability (P < 0.05). A negative correlation (r = -0.70***) was found between the side of the temporomandibular joint sound and the side of unilateral contact in RCP. Boys with unilateral contacts in RCP had more non-reciprocal clicking than girls. No signs were found indicating that a unilateral contact in RCP is an aetiological factor for development of temporomandibular disorders. Unilateral contacts in RCP may in adolescents be considered a predictive factor for temporomandibular joint disturbance. Contradictory causes may determine the sagittal distance between RCP and ICP. PMID- 7472733 TI - The importance of a multidisciplinary approach to the assessment of patients with obstructive sleep apnoea. AB - Obstructive sleep apnoea may present with a wide range of symptoms resulting in a variety of referral pathways. A multidisciplinary approach to examination and diagnosis helps to determine the most appropriate treatment plan for each individual. The subject is seen by each member of the team, appropriate investigations undertaken and a further meeting arranged at which all opinions are discussed. A reasoned treatment regime is produced, taking into consideration the patient's wishes and overall medical condition. This paper describes the team approach currently employed in the Department of Thoracic Medicine at The Prince Charles Hospital, Brisbane, Australia. The thoracic physician and ENT surgeon work in close collaboration with their dental colleagues: an orthodontist, prosthodontist and a maxillofacial surgeon. An outline of the examination and investigations made by each is described and the multidisciplinary approach is illustrated by a description of the management of five subjects with suspected obstructive sleep apnoea. PMID- 7472734 TI - Temporomandibular disorders in the active phase of orthodontic treatment. AB - Subjective symptoms and clinical signs of temporomandibular disorders (TMD) as well as presence of headache, bruxism and occlusal interferences were examined in 50 patients (mean age 12.9 years) before, during and immediately after orthodontic treatment. The prevalences of signs and symptoms of TMD were high before the treatment in comparison with subjects in an epidemiological study. Except for TMJ sounds, signs and symptoms of TMD as well as presence of headache decreased during the treatment. The major factor for decrease of the dysfunction index during the treatment compared with before the treatment, was tenderness to palpation of the masticatory muscles. Although there was a high prevalence of occlusal interferences during treatment, they seemed to have little importance for development of TMD. One explanation may be that the orthodontically moved teet are sensitive to contact resulting in a decrease of oral parafunctions. Such a decrease was also reported by the patients. PMID- 7472735 TI - Ultrasound imaging of the masseter muscle in myotonic dystrophy patients. AB - Ultrasound technique was applied to measure the thickness and examine the internal structure of the masseter muscle in a group of 16 adult patients (nine women and seven men) with myotonic dystrophy (MyD) and 16 healthy individuals matched in age, sex and number of occluding teeth. The masseter thickness was measured bilaterally under both relaxed conditions and during maximal clenching. The error of measurement was found to be small, not exceeding 0.45 mm. The imaging characteristics of the masseter in most of the MyD patients was an obvious atrophy of the muscle with increased echointensity of the intramuscular tissue and loss of the internal structure concerning tendons and fasciae. The mean masseter thickness (+/- SD) in the MyD group was 10.4 (+/- 2.2) mm under relaxed conditions and 11.1 (+/- 2.4) mm during maximal clenching, compared with 13.3 (+/- 2.2) mm and 14.1 (+/- 2.4) mm, respectively, in the healthy group (P < 0.001). In conclusion, our results indicate that, in most of the myotonic dystrophy patients, the masseter muscle is atrophic with obvious signs of degeneration. Ultrasound is a useful method for both qualitative and quantitative evaluation of the condition of the masseter muscle. PMID- 7472736 TI - Anatomy of the moose craniomandibular joint (Alces alces, Linnaeus, 1758). AB - the craniomandibular joints (CMJ) were investigated in 10 Swedish mooses (eight cows: five 1-year-old, two two-year-old, and one 7-year-old) together with one bull moose (2 years) and a yearling. The CMJs, situated at a high distance from the occlusal plane of the tooth rows, comprised a temporal component, a disc and a condyle. The temporal component lacked a mandibular fossa as well as articular eminence. The joint surface was oval, almost flat and convex. The condyle had about two-thirds of its length medio-laterally, cylindrical in shape and from a posterior view medially inclined. The articulating surface was divided into a horizontal part located on the superio-anterior condyle, and a vertical part medially situated. The disc was oval in shape and revealed two synovial chambers totally surrounded by a soft tissue lining. PMID- 7472737 TI - Bite force and temporomandibular disorder in juvenile chronic arthritis. AB - The aim of this study was to investigate the functional condition of the stomatognathic system in children suffering from juvenile chronic arthritis, with respect to bite force and temporomandibular disorder in relation to radiographic abnormalities of the mandibular condyle, occlusal factors and systemic disease parameters. Thirty-five children with juvenile chronic arthritis were compared to 89 healthy children with an Angle Class I occlusion and 62 children with an Angle Class II malocclusion. Subjective symptoms and clinical signs of temporomandibular disorder and radiographic mandibular condylar changes were more common in children with juvenile chronic arthritis than in the two comparison groups. Maximal molar and incisal bite forces and maximal molar bite force endurance times were also significantly reduced in children with juvenile chronic arthritis. It is concluded that the differences between the groups are caused mainly by the systemic inflammatory disease itself, but a functional influence of weakened masticatory muscles cannot be excluded. PMID- 7472738 TI - Relationships between facial morphology and activity of orofacial muscles in patients with a complete upper and a partial lower denture. AB - In a sample of 30 partially edentulous subjects, provided with an immediate complete upper denture and a partial lower denture and followed for 2 years, a correlation analysis was performed between facial morphology, as measured by lateral head films, and EMG peak mean voltages of the anterior temporal and masseter muscles when clenched and during swallowing and chewing, and of the lip muscles during swallowing. At the pre-treatment stage, when the patients occluded on a residual anterior dentition, correlations between facial morphologic characteristics and peak mean voltages of the jaw closing muscles indicated stronger muscle activity in individuals with a square facial type. No significant associations were observed between muscle activity and the age of the subjects, which ranged from 36 to 71 years. After placement and use of the dentures a significant forward-upward rotation of the mandible with a decrease in overbite occurred, as a result of resorption of the edentulous maxillary ridge. At the 1 year stage of denture use, the elevator activity showed no significant correlations with facial morphology. On the other hand, negative correlations were observed especially between the anterior temporal activity and anterior overbite. Similar negative correlations were observed for the lip activity during swallowing, especially of the upper lip. These findings suggest occlusal contact between the incisors during the various functions in subjects with a small overbite. The findings further suggest that the anterior temporal and the upper lip muscles are sensitive to changes in anterior occlusal relations of the dentures. PMID- 7472739 TI - Dental state and functional capacity in 75-year-olds in three Nordic localities. AB - The aims of the study were to compare dental state in three Nordic 75-year-old populations and to evaluate dental state as a marker of functional ageing. The study is part of a Nordic comparative study of 75-year-olds in Glostrup, Goteborg and Jyvaskyla. In the dental studies, 411 subjects in Denmark, 308 in Sweden and 310 in Finland participated. Odontological history was obtained from interviews. Other methods, which are described elsewhere, were used to measure various functional parameters. In Goteborg, 23% of the 75-year-olds reported they were edentulous and 27% that they had more than 20 of their own teeth. The corresponding figures in Glostrup were 45% and 15% and in Jyvaskyla 58% and 9%, respectively. In all three localities, economic factors and lifestyle factors, such as tobacco smoking, low physical activity and low social activity, were significant predictors for impaired dental health. The number of teeth was significantly associated with, for example, spirometry parameters, reaction time, body extention and body flexion in males. In females, significant associations were obtained with spirometry parameters, reaction time, handgrip, body extension and body flexion. Internordic differences regarding dental state were obvious. Dental state was associated with functional capacities. We suggest that dental state is a marker of functional capacity in elderly populations. PMID- 7472740 TI - Implant prosthodontic treatment options for the edentulous patient. AB - Longitudinally documented benefits for edentulous patients of treatment outcomes of the osseointegration technique demonstrate compelling therapeutic results. Heterogeneous population groups, treated in different centres by various dental specialists, have provided impressive evidence of a minimal burden of illness associated with the procedure. A critical appraisal of the many components of any implemented treatment paradigm demands repeated analysis of the factors which enable patients' informed consent and dentists' optimal decision making. This paper reviews those salient aspects which impact upon decision making with implant-supported prostheses. It emphasizes the predominance of bone structure in selecting the most likely favourable treatment outcome. PMID- 7472741 TI - Comparison of the type-2 insulin-like growth factor receptor in normal osteoblasts and osteosarcoma-derived osteoblast-like cells. AB - Insulin-like growth factor-II is known to stimulate the proliferation and differentiation of osteoblasts in part through activation of the type-2 insulin like growth factor receptor. The present study examined the type-2 insulin-like growth factor receptors of three normal osteoblast-like cells and three osteosarcoma-derived osteoblast-like cells (OGA, SU, and IMAI) from humans. [125I]insulin-like growth factor-II was used for the binding studies. All of the cell types had high affinity binding sites for insulin-like growth factor-II (dissociation constants [Kd] < or = 1 nM). The concentration of these sites was 10 to 24-fold higher in normal osteoblasts than in the osteosarcoma cells studied. Unlabeled insulin-like growth factor-II inhibited the binding of [125I]insulin-like growth factor-II to the cells in a dose-dependent manner; however, unlabeled insulin-like growth factor-I and insulin were less effective. Covalent crosslinking of insulin-like growth factor-II binding sites gave molecular mass estimates of M(r) 250,000 in human osteoblast cells, 250,000 and 130,000 in OGA cells, 240,000 in SU cells, and 250,000 and 130,000 in IMAI cells. Unlabeled insulin-like growth factor-II inhibited all affinity labeling. In Northern blot analysis, the type-2 insulin-like growth factor receptor mRNA of normal osteoblasts was seen in greater abundance than it was in osteosarcoma cells. These results indicate that the numbers of type-2 insulin-like growth factor receptors differ between normal and transformed osteoblasts and that the differential expression of the receptor may be due to the differentiation of osteoblasts. PMID- 7472742 TI - De novo synthesis of type-I collagen in bone biopsy material. AB - A simple and rapid method was established for the cultivation of bone cell tissue. Human bone tissue derived from orthopaedic surgery was cultivated in the presence of 14C-proline and beta-aminopropionitrile. De novo synthesized collagen was extracted from the tissue and quantified by determination of radioactivity in the purified protein. Measurements of the oxygen consumption of the tissue provided evidence that the physiological conditions for the tissue were optimal. The tissue was vital over a period of as long as 7 days, showing normal respiration and a constant rate of collagen synthesis. The observed levels of alkaline phosphatase and acid phosphatase activity clearly demonstrated that mainly osteoblasts were involved in metabolic activity. The described system is suitable for investigations of bone cell metabolism under quasiphysiological conditions. PMID- 7472744 TI - Proliferative and phenotypic responses of bone-like cells to mechanical deformation. AB - Limited in vivo and in vitro experiments suggest that bone and bone-like cells respond to mechanical signals in a trigger-like rather than a dose-response fashion; i.e., they fail to respond until they have been stimulated with some given number of cycles of loading, and then once they respond, additional cycles produce little or no effect. To explore this notion, rat calvaria-derived osteoblast-like cells and the cell line MC3T3-E1 were plated at a high cell density (5,000 cells/mm2) on silicone membranes coated with type-I collagen and were allowed to attach for 24 hours. The membranes then were exposed to vacuum pressure (-1 kPa, 0.5 Hz) on a daily basis, and cultures were assayed every 2 days for 2 weeks. The proliferation of nontransformed cells increased 7-fold with as few as four daily cycles but not with one cycle per day. Furthermore, 1,800 cycles of vacuum did not result in a greater response than four cycles per day. We observed inverse phenotypic responses: the expression of osteocalcin was depressed compared with controls in the cultures of osteoblast-like cells that were strained with as few as four cycles per day. Alkaline phosphatase activity was depressed in the cultures of both the osteoblast-like cells and the MC3T3-E1 cells exposed to low vacuum pressures (-1 kPa) with four daily cycles of vacuum pressure. Increasing the vacuum magnitude did not affect the occurrence of a "trigger response" between one and four cycles of vacuum application.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7472743 TI - Involvement of platelets in stimulating osteogenic activity. AB - Osteoblast-like cells have been shown to be sensitive to the proliferative action of a wide variety of growth factors. Many of these growth factors have been isolated from platelets and are thought to be released at local sites in response to injury. In this study, we tested whether human platelet concentrate, as a supplement to basic medium, would support the proliferative and functional activity of human fetal osteoblast-like cells in both short-term and long-term culture. In short-term studies, uptake of [3H]thymidine was increased in platelet treated cultures by more than 4-fold compared with 10% serum-supplemented controls. When cultured for prolonged periods on coverslips, the cells formed multilayers, with a collagen-based matrix separating the layers. Long-term cultures that were treated with 1.5% (vol/vol) platelets in serum-supplemented medium showed increases in the depth of the multilayers of as much as 36-fold at 30 days after confluence, compared with the 10% serum-supplemented controls; this difference persisted until day 50. Incorporation of growth factor in the matrix was examined with the use of colloidal gold immunoelectron microscopy. Immunogold labeling intensities for transforming growth factor-beta 1 were significantly lower in the platelet-treated cultures at 20 days and then increased to a maximum level of 2.1-fold more than in the controls at 40 days. Labeling intensities for insulin-like growth factor-I and basic fibroblast growth factor were significantly lower in the platelet-treated cultures than in the controls at all stages of culture.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7472745 TI - Osteosarcoma oncogene expression detected by in situ hybridization. AB - Fifteen archival human osteosarcoma specimens were examined by in situ hybridization for the expression of human and mouse transforming growth factor beta (isoforms 1, 2, and 3), c-fos, and metalloproteinase (stromelysin-3 and matrilysin). Osteosarcoma subtypes were confirmed by review of patients' radiographs, histopathology, and age at diagnosis. The outcome and method of treatment were documented. The subtypes of osteosarcoma consisted of nine conventional osteosarcomas and two each of fibroblastic, telangiectatic, and post radiation osteosarcomas. Each specimen was histologically examined under light microscopy, and then adjacent paraffin sections were assayed with sense and anti sense RNA probes by in situ hybridization. The probes localized to the neoplastic cells, confirming the methodology of the technique. Human transforming growth factor-beta 1 had the most uniform binding affinity to the osteosarcomas examined and was more specific in binding than mouse transforming growth factor-beta 1. Specific mRNA encoding for the transforming growth factor-beta s, c-fos, and metalloproteinases are detectable in patterns within osteosarcoma cells, and collectively, their expression parallels the different histopathologic subtypes. The less differentiated subtypes (telangiectatic and post-radiation osteosarcomas) expressed the fewest molecular markers. Osteosarcoma is a heterogeneous tumor. Differential expression of matrilysin in osteosarcoma is the first reported detection of metalloproteinase activity in human skeletal sarcoma. PMID- 7472746 TI - Incorporation of polylactide-polyglycolide in a cortical defect: neoangiogenesis and blood supply in a bone chamber. AB - Erodible polymers are an alternative to metals for fracture fixation (for example, in the malleolus) and for maxillofacial reconstruction. In this study, the vascular response to eroding polylactide-polyglycolide copolymer threads was observed chronically in a bone chamber implant, with use of intravital microscopy. A bone chamber implant loaded with 100 microns thick polylactide polyglycolide threads was implanted into the right tibia in 15 mature female New Zealand White rabbits. Periodic intravital microscopic observations were performed from the third to the tenth or twelfth week after implantation. Vascularization, blood flow, and trabecular growth into the chambers from the medial cortex were recorded on videotape and analyzed using digital image processing. A statistically significant delay of neo-osteogenesis in the presence of this copolymer was described in an earlier report. The present report describes the measures of neoangiogenesis and blood supply; there was a significant delay in neoangiogenesis. It is suggested that both delayed angiogenesis and osteogenesis were secondary consequences of the macrophage response to slowly eroding poly-L-lactide crystal nanoparticles and the influence of reduced nutrient exchange. The lesser effect on blood supply and vascular volume fraction was seen to be linked to the slowing down of angiogenesis, as the latter allowed vessels to mature, with a widening of their calibers. This homeostatic adjustment was interpreted as being only partially successful in restoring control levels of oxygen delivery, because resulting increases in vessel surface area did not reach control levels. Thus, in the presence of eroding polylactide-polyglycolide, the oxygen supply and extravasation of other nutrients may be below normal during healing phases when the need is critical. PMID- 7472747 TI - Three-dimensional geometric and structural symmetry of the turkey ulna. AB - Structural models of long-bone preparations usually assume left-right symmetry of contralateral bones under normal (baseline) conditions. To obtain insight on how this assumption affects the detection of subtle changes (as from functional adaptation), we formally examined the three-dimensional geometric and structural symmetry of paired long bones, using contemporary image reconstruction and stress analysis techniques. Nine pairs of ulnae from normal male turkeys were reconstructed computationally from serial transverse images obtained by either (a) mechanical sectioning and digital photographic imaging or (b) computed tomography. Computed tomography scans allowed greater precision in reconstruction than did digitally imaged photographs. Left-right comparisons of parameters of geometric symmetry (from computed tomography reconstructions) revealed average differences in whole bone volume and whole bone principal moments of inertia of 3.6 and 3.0%, respectively. Differences in bone curvature were indexed as noncolinearity of left compared with (mirrored) right centroidal axes, giving a disparity of 0.7 +/- 0.3 mm. Within the longitudinal central 20% of the diaphysis (the customary region for histomorphometry), average left-right differences in cross-sectional area and area principal moments of inertia for computed tomography images were 4.7 and 5.0%, respectively. The overlap of longitudinally paired cross sections of the mid-diaphysis, aligned at common centroids and oriented in the respective principal inertial directions, was greatest (as much as 95%) in the central 20% of the diaphysis. Paired three-dimensional finite element models demonstrated nearly identical left and right stress/strain fields throughout the ulnar diaphyses for both compressive and torsional loading. Our data suggest that the assumption of contralateral geometric symmetry in long bones should be judged in the context of the specific attribute of symmetry under consideration; however, we conclude that for purposes of finite element modeling the assumption of symmetry is reasonable. PMID- 7472748 TI - Hindlimb suspension diminishes femoral cross-sectional growth in the rat. AB - Growth, functional adaptation, and torsional strength were examined in the femora of 39-day-old male Sprague-Dawley rats subjected to hindlimb suspension for 0, 1, 2, 3, or 4 weeks and were compared with measurements for age-matched control animals. Our goal was to understand the effect of reduced loading on the normal age-related changes in femoral properties during growth. The control animals exhibited growth-related increases in all geometric and torsional properties of the femur. The mean body mass and femoral length of the hindlimb-suspended rats were similar to those of the controls throughout the experiment. Over 4 weeks, the femoral cross-sectional and torsional measurements from the hindlimb suspended rats demonstrated increases in comparison with the basal values (+33% cross-sectional area, +64% polar moment of inertia, +67% ultimate torque, and +181% torsional rigidity), but the age-matched controls showed significantly greater growth-related increases (+71% cross-sectional area, +136% polar moment of inertia, +127% ultimate torque, and +367% torsional rigidity). The differences in femoral structural strength between the hindlimb-suspended animals and the age matched controls were attributable to differences in altered cross-sectional geometry. PMID- 7472749 TI - Uniformity of resorptive bone loss induced by disuse. AB - Strains induced in the skeleton by functional activity are critical to the homeostasis of bone tissue. An in vivo model of disuse osteopenia was used to examine whether the removal of these regulatory stimuli induces a uniform loss of cortical bone through the whole organ or whether the loss of bone is focused at specific sites of the cortex. The right radii of five adult male turkeys were isolated from their normal functional loading for 8 weeks. The corresponding left radius from each animal served as an intact contralateral control. An additional group of five turkeys was used as time-zero controls to assess the initial areal symmetry of the left and right radii. Areal properties were assessed at three sites at equal intervals spanning the middle 3 cm of the diaphysis. Adaptation was determined for each cross section as a whole, as well as specifically by site by division of each cross section into 12 equal angle sectors. The average across all experimental sections after 8 weeks of disuse was 12.1 +/- 1.9% (+/- SE) loss of bone mass. The change in mean cross-sectional area varied little between the three diaphyseal sites (-10.2 +/- 3.3%, -13.5 +/- 3.8%, and -12.6 +/- 4.0%) and occurred primarily (84%) by uniform expansion of the endosteal envelope. However, elevated intracortical porosity following 8 weeks of disuse was highly nonuniform, with 58% of the increased porosity preferentially located in the ventral/caudal cortex (representing only 25% of the cortical area).(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7472750 TI - Cortical bone perfusion in plated fractured sheep tibiae. AB - The limited contact dynamic compression plate and partial contact plate were designed to decrease contact with cortical bone in an attempt to decrease cortical ischemia, remodeling, and eventual porosis under the plate after use of standard dynamic compression plates. This study quantified cortical bone blood flow beneath the plate with these three different designs in a sheep tibia fracture model. In 18 skeletally immature sheep, the right tibia was fractured and then was internally fixed with an interfragmentary screw and a dynamic compression plate, limited contact dynamic compression plate, or partial contact plate. At 12 weeks, cortical bone perfusion was assessed with laser Doppler flowmetry in nine areas beneath the plate. The baseline (before fracture) cortical bone cell flux averaged 100 +/- 60 mV. After fracture, this decreased to 60 +/- 48 mV (p < 0.0003); immediately after plating, the perfusion averaged 29 +/- 25 mV (p < 0.01). Cortical bone perfusion then increased to 106 +/- 52, 165 +/- 71, and 163 +/- 71 mV at 2, 6, and 12 weeks after fracture (p < 0.001 for all when compared with values after plating). No significant differences in cortical perfusion were seen between the types of plate. Cortical porosity under the plate was assessed with digital density analysis of microradiographs of this region. No significant difference was seen between the types of plate in this analysis or in biomechanical and disulphine blue perfusion analysis. Thus, no significant advantage was seen for the new plate designs used in this model. This lack of advantage may be a result of the immature animals used in the study, the protocol for blood flow measurement, the invasive periosteal stripping employed to create the fracture, or all three. However, as advantages with the new plate designs have been seen in other studies, this area warrants further investigation. PMID- 7472751 TI - Evaluation of influence of 24-hour cold preservation on endothelin production and on endothelin receptors in the bone vasculature. AB - Endothelin-1 is a vasoactive peptide produced by the vascular endothelium. It is one of the most potent endogenous vascular smooth muscle constrictors. Two subtypes of the endothelin receptor have been cloned and sequenced and denoted endothelin-A and endothelin-B. The aim of this study was to define the influence of cold ischemia on the production of endothelin-1 and on the endothelin receptors. Two different preservation techniques (cold storage only and cold storage with microperfusion with University of Wisconsin solution) also were compared. The study was performed in an in vitro bone perfusion model to isolate the vascular endothelium from blood components. The production of endothelin-1 by the bone vasculature was not altered after 24 hours of cold ischemia. No contractions were observed with S6c, a selective endothelin-B agonist, and this effect was not influenced by cold ischemia. The response mediated by the endothelin-A receptor was increased significantly, an effect that was not influenced by preservation with University of Wisconsin solution. This latter finding was the only significant alteration in the vascular function detected in the in vitro model after 24 hours of cold ischemia. With regard to the pharmacologic properties of endothelin-1, this mediated response could be implicated in the pathogenesis of vasospasm. PMID- 7472752 TI - Correlation between synthetic activity and glycosaminoglycan concentration in epiphyseal cartilage raises questions about the regulatory role of interstitial pH. AB - Current data provide compelling evidence that the pH of the interstitial fluid of cartilage is an important determinant of the metabolic activity of chondrocytes, and this has served as the basis for a mechanistic proposal whereby chondrocytes could sense mechanical compression. The objective of the current study was to test this hypothesis further by examining biosynthetic activity in cartilage as a function of glycosaminoglycan content, which is the major determinant of interstitial pH. On the basis of previous data, increased biosynthetic activity would be anticipated to correlate with a decreased glycosaminoglycan content and an elevated interstitial pH. In contrast to our expectations, we found that the biosynthetic activity (monitored by measurement of incorporation of sulfate and proline) was positively correlated with the glycosaminoglycan content of tissue. These results raise doubt as to whether interstitial pH provides a dominant mechanism for controlling the metabolism of chondrocytes. PMID- 7472753 TI - Time-dependent changes in the response of cartilage to static compression suggest interstitial pH is not the only signaling mechanism. AB - The goal of the present study was to reexamine the role of interstitial pH in regulating the biosynthetic rate in cartilage tissue by addressing two research questions: (a) Do small, short-term changes in interstitial pH, induced independently by two different mechanisms (namely, by controlling the pH of the medium or by mechanical compression), result in biosynthetic rates commensurate with those expected from the "natural" relationship between interstitial pH and biosynthesis? and (b) Are the effects of changes in the pH of the medium or in compression the same for short-term (14-hour) and long-term (60-hour) exposures? Biosynthetic rates were estimated from incorporation of sulfate and proline into explants of bovine epiphyseal cartilage during the final 14 hours of culture. These rates decreased with decreasing pH of the medium, with increasing compression, and with decreasing native glycosaminoglycan content; or, expressed in terms of interstitial pH, acidification induced by compression or by lowering the pH of the medium resulted in a decreased biosynthetic rate, whereas interstitial acidification effected by increasing glycosaminoglycan content enhanced it. When the time for which tissue was exposed to changes in the pH of the medium was increased from 14 to 60 hours, the relationship between the biosynthetic rate and the pH remained constant whereas the relationship between the biosynthetic rate and compression was reversed. These data suggest that the transduction mechanisms underlying the response to pH of the medium and compression differ and that some adaptation or stimulation by modest levels of compression can occur with longer exposures. Interstitial pH is not the sole determinant of biosynthesis, and it cannot really account for the long-term response of cartilage tissue to static compression. PMID- 7472754 TI - Integrative repair of articular cartilage in vitro: adhesive strength of the interface region. AB - The objective of this study was to quantify the strength of the repair tissue that forms at the interface between pairs of cartilage explants maintained in apposition in an in vitro culture system. Articular cartilage explants were harvested from calves and from adult bovine animals, dissected into uniform blocks, and incubated in pairs within a chamber that maintained a 4 x 5 mm area of tissue overlap. Following 1-3 weeks of incubation, integrative repair was assessed by testing samples in a tensile single-lap configuration to estimate adhesive strength. After incubation in medium containing 20% fetal bovine serum, the adhesive strength between pairs of calf cartilage blocks and pairs of adult bovine cartilage blocks increased at a rate of 7.0 and 10.5 kPa/week, respectively. This repair process appeared to be dependent on viable cells, since lyophilization of adult bovine cartilage before incubation completely inhibited the development of an interface with a measurable adhesive strength. The repair process was dependent on serum components in the medium. Incubation of sample pairs for 3 weeks in medium supplemented with 20% fetal bovine serum resulted in a relatively high proteoglycan content as well as a relatively high adhesive strength (34 kPa), whereas incubation in basal medium with or without 0.1% bovine serum albumin resulted in a 54-70% lower proteoglycan content and a 65-88% lower adhesive strength. Samples incubated for 3 weeks with serum also had a 20% higher DNA content than samples maintained in basal medium. Histological analysis indicated some cell division at the free surfaces of the explant and also occasional cells within the interface region between explants. PMID- 7472755 TI - Transforming growth factor-beta 1 and fibroblast growth factors in rat growth plate. AB - Chondrocytes in the growth plate progress in an orderly fashion from resting through proliferating to hypertrophic cells. In the region of hypertrophic chondrocytes, the cartilage is invaded by capillary loops and endochondral ossification is initiated. It is currently believed that growth factors may regulate the proliferation and maturation of chondrocytes and the synthesis of extracellular matrix in the growth plate. The ordered sequence of proliferation and differentiation observed in the growth plate provides a unique opportunity to study the role of acidic fibroblast growth factor, basic fibroblast growth factor, and transforming growth factor-beta 1 in the regulation of these processes. In this study, expression of the mRNA of these growth factors was examined using total RNA extracted from the physis and epiphysis of rat tibias. Transforming growth factor-beta 1 mRNA was detected by Northern hybridization. Expression of the genes encoding acidic and basic fibroblast growth factors was demonstrated by polymerase chain reaction amplification. In addition, using polyclonal antibodies against these growth factors, we localized them by immunohistochemical analysis. Strong intracellular staining with a predominantly nuclear pattern was observed in chondrocytes from the proliferating and upper hypertrophic zones. In contrast, chondrocytes in the resting zone stained only faintly for the presence of these growth factors. Some chondrocytes in the resting zone adjacent to the proliferating zone stained with these antibodies, and the antibodies also stained cells in the zone of Ranvier, which regulates latitudinal bone growth. Lastly, the location of transforming growth factor-beta 1 was examined further with use of a polyclonal antipeptide antibody specific for its extracellular epitope.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7472756 TI - Primary osteoarthrosis in guinea pigs: a stereological study. AB - Volumes and surfaces of articular cartilage and subchondral bone of the proximal tibial epiphysis were measured by unbiased stereological methods on the light microscopic level in groups of 6, 12, and 30-month-old (adult, middle-aged, and old) guinea pigs with primary osteoarthrosis. At 12 months, structural changes similar to those of human osteoarthrosis had developed, predominantly on the central medial condyle, which was not covered with meniscus. The lateral condyle was virtually unaffected; this allowed separate analysis of age-related and disease-related changes. Fibrillation and destruction of cartilage was accompanied by a simultaneous increase of the volume of both cartilage (66%) and subchondral bone (50%). The epiphyseal volume increased by 27% at 12 months, predominantly on the medial (osteoarthrotic) condyle, whereas the volume of the lateral condyle increased only in the oldest age group; this indicated that the joint has a potential for growth and remodeling. Joint growth has been suggested as a pathogenic factor in osteoarthrosis. The 65% increase in thickness noted in subchondral bone was a further indication of the importance of proliferative bone changes in early osteoarthrosis. The low variability of osteoarthrosis in this animal model makes it possible to obtain stable quantitative data from relatively small groups of animals. PMID- 7472757 TI - Arthritis not immobilization causes bone loss in the carrageenan injection model of inflammatory arthritis. AB - One suggested cause of the high turnover osteopenia of experimental inflammatory arthritis is disuse of affected joints. To compare the influence of immobilization or disuse, or both, with that of inflammatory arthritis on bone turnover, rabbits were placed into four groups. In group 1, arthritis was induced in the right knee by seven intra-articular injections of 1% carrageenan, over 49 days; in group 2, a plaster cast was applied to immobilize the right hindlimb in flexion; in group 3, arthritis was induced and the hindlimb was immobilized; and in group 4, nothing was done (control). The fluorescent label calcein was administered in drinking water (0.05%) ad libitum to all groups on days 22-36. On day 49, specimens were prepared for analysis of bone volume and new bone volume at a near site (right femur) and at remote sites (contralateral femur and ipsilateral humerus). The data were analysed by multiple regression and Bonferroni tests. In group 1, new bone volume was three times higher than in group 2 or 4 (p < 0.05 for each comparison); this indicated increased bone remodeling in the right femur. This contrasted with group 2, in which neither index of bone remodeling was changed. The combination of immobilization with arthritis resulted in more intense osseous effects of inflammatory arthritis, with a one-quarter decrease in bone volume (group 3, 30.99 +/- 2.50; group 4, 42.07 +/- 2.38, p < 0.05), as well as a 4-fold increase in new bone volume (p < 0.001) compared with group 1.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7472758 TI - Laxity characteristics of normal and pathological murine knee joints in vitro. AB - The aim of this study was to validate a device developed previously to measure laxity of murine knee joints and to investigate whether experimentally induced pathological conditions result in measurable laxity. The laxity characteristics of normal murine knee joints were derived from measurements of 25 left knees of normal mice. Reproducible, nonlinear s-shaped load-displacement curves were determined, and parameters of anterior-posterior translation, varus-valgus rotation, and compliance were calculated from the curves. No differences were found between the left and right knee joints of eight mice. The average displacement between 0.8 N of anterior force and 0.8 N of posterior force was 0.47 +/- 0.10 mm. The endpoint compliances for anterior and posterior displacements were 0.16 +/- 0.03 and 0.16 +/- 0.04 mm/N, respectively. The average rotation between a 4 Nmm valgus moment and a 4 Nmm varus moment was 17.4 +/- 3.3 degrees. The endpoint compliances for varus and valgus rotations were 1.1 +/- 0.7 and 1.0 +/- 0.3 degrees/Nmm, respectively. Storage of the joints at -70 degrees C had no effect on laxity. We also studied the parameters of laxity after pathology of the knee joint was induced. Zymosan-induced or antigen-induced arthritis did not increase laxity of the joint. In an osteoarthritis model induced by injection of collagenase, laxity was markedly increased. In conclusion, laxity in the knees of mice can be measured reproducibly and changes in the characteristics of laxity due to pathological conditions can be quantified. PMID- 7472759 TI - Human monocyte response to particulate biomaterials generated in vivo and in vitro. AB - We studied the ability of four clinically relevant particle species to stimulate human peripheral blood monocytes to release bone-resorbing agents, including interleukin-1 (both interleukin-1 alpha and interleukin-1 beta), interleukin-6, and prostaglandin E2. The species studied were titanium-6% aluminum-4% vanadium (TiAlV), commercially pure titanium, fabricated ultrahigh molecular weight polyethylene, and polyethylene retrieved from interfacial membranes of failed uncemented total hip arthroplasties. For all species, the mean size was less than 1 micron. Human peripheral blood monocytes were challenged with these particles in a uniform manner on the basis of surface area. Phorbol 12-myristate acetate, zymosan, and nonphagocytosable titanium particles served as controls. Stimulation of human monocytes is a function of the composition and concentration of particles. In this study, TiAlV particles appeared to be the most competent to elicit the synthesis and release of inflammatory mediators. Particles of commercially pure titanium and of fabricated ultrahigh molecular weight polyethylene also could induce the release of various cellular mediators, albeit at a lower level, whereas the particles of polyethylene retrieved from interfacial membranes were less stimulatory in these short-term in vitro experiments. PMID- 7472760 TI - Role of muscles in lumbar spine stability in maximum extension efforts. AB - Many problems of the lumbar spine that cause pain are attributed to instability. The ligamentous spine (without muscles) is unstable at very low compressive loads. This study examined the hypothesis that instability of the lumbar spine is prevented under normal circumstances by the stiffness of spinal musculature, without active responses from the neuromuscular control system. The effect of muscle activity (force and stiffness) on the stability of the lumbar spine was analyzed for maximum voluntary extension efforts with different spinal postures in the sagittal plane. The analysis included realistic three-dimensional representation of the muscular anatomy with muscles crossing several motion segments. The stiffness of motion segments was represented using in vitro measured properties. Under a range of conditions with maximum extension effort, active muscle stiffness was required to prevent the lumbar spine from buckling. The dimensionless value of the muscle stiffness parameter q as a function of activation and length had to be greater than a critical value in the range of 3.7 4.7 in order to stabilize the spine. Experimentally determined values of q ranged from 0.5 to 42. These analyses demonstrate how changes in motion segment stiffness, muscle activation strategy, or muscle stiffness (due to degenerative changes, injuries, fatigue, and so on) might lead to spinal instability and "self injury." PMID- 7472761 TI - Frozen storage increases the ultimate compressive load of porcine vertebrae. AB - The use of freezing as a method of storage is commonplace in mechanical testing of biological tissues. The effects of freezing on tissues that comprise spinal segments have been examined separately, but little work has been done on intact specimens. We examined the effect of freezing on the structural properties of porcine cervical spines. The intact cervical spines of seven pigs (a total of 14 specimens--seven of C2-C4 and seven of C5-C7) were stored frozen (-20 degrees C) for 1 month. The ultimate compressive load, displacement, stiffness, and energy absorbed were obtained using a monotonic compressive load applied at 3,000 N/sec. The structural properties were compared with those of another 14 porcine cervical specimens (control group, matched for age and weight) that were tested in a fresh state. The frozen storage of the vertebral specimens significantly increased the ultimate compressive load (24%) and energy absorbed to failure (33%). The stiffness and displacement at failure were not affected. We concluded that the use of freezing as a storage medium should be of concern when the resulting measures are used to quantify the ultimate compressive load of the spinal motion segments. PMID- 7472762 TI - [Audiologic evaluation in a family showing diabetes mellitus and deafness associated with a mutation in mitochondrial DNA]. AB - An A to G transition at nucleotide 3,243 in the tRNA(Leu)(UUR) gene of mitochondrial DNA has recently been identified as a pathogenic point mutation which is associated with diabetes mellitus and sensorineural deafness in several pedigrees. We have also reported a family showing the association of deafness and diabetes mellitus as the predominant clinical features with this mutation. Audiologic data from two patients in this family are presented. Both had a bilaterally symmetrical sensorineural hearing loss at all frequencies. As is often the case with deafness associated with a mitochondrial disorder, the pure tone threshold values were maximal at high frequencies in both patients. The audiologic work-up presented not only cochlear characteristics but also signs suggestive of retrocochlear disturbance with poor speech discrimination scores as compared to pure-tone thresholds, although auditory brain-stem responses showed neither wave delay nor prolonged interpeak latencies. PMID- 7472763 TI - [Efficacy and complications of topical cocaine anesthesia in functional endoscopic sinus surgery]. AB - A 4% cocaine (total dose; 200 mg) solution with 1 mg epinephrine was used as a topical anesthetic for 178 cases undergoing functional endoscopic sinus surgery. Complications were investigated before and during surgery. Bleeding amount and operative duration were compared between patients with and without topical cocaine application. Tachycardia (26 cases) was the only complication encountered in this study. Tachycardia occurred at a high incidence in cases which used atropine as a premedication. Total bleeding amount did no differ significantly between those with and without topical cocaine. However, minimum bleeding cases (less than 20 ml) were found in the cocaine group more frequently than in the non cocaine group. Operative durations of cases with cocaine anesthesia were significantly shorter than cases without cocaine anesthesia. Those findings suggest that cocaine is effective for prevention of bleeding from micro-vessels and/or mucosal oozing, whereas bleeding from macro-vessels was not controlled by topical cocaine application. Topical cocaine anesthesia produced no major complications, even at 200 mg, in the presence of 1 mg epinephrine and when atropine use was avoided. PMID- 7472764 TI - [Usefulness of three-dimensional CT angiography for cervical tumor]. AB - In this study, the usefulness of three-dimensional angiographic imagings from helical scanning computed tomography (CT) enhanced by contrast media were evaluated based on results from three patients with a cervical mass, e.g. thyroid cancer and carotid body tumor. Three-dimensional CT (3D-CT) angiographic imagings in these cases clearly demonstrated secondary dislocation or distortion of the major cervical vessels caused by the tumor or metastatic lymph nodes. It was remarkable that in the highly vascularized tumor in case 3, all features of the tumor were delineated by 3D-CT angiography. Our results indicate that 3D-CT angiographic imagings are especially useful for preoperative evaluation of the relationship of the cervical mass to major cervical vessels as well as in the observation of tumors rich in vasculature. PMID- 7472765 TI - [The volume ratio between squamosal and petrosal air cells divided by Korner's septum]. AB - Pneumatized air cells of the temporal bone are embryologically divided into two portions; the squamosal portion and the petrosal portion. These two portions are anatomically separated by the petrosquamous fissure and petrosquamous lamina, the so-called Korner's septum. In this study, the volumes of the two portions were measured with Target Imaging CT in 5 normal ears (3 subjects) with a clearly recognizable Korner's septum. The combined volume of the two portions, in all CT slices, was also measured. The squamosal portion occupied 32.7% of the total air cell volume, on average, and the petrosal portion was 67.3%. A positive correlation was observed between the total air cell volume of the temporal bone and the volume ratio of the petrosal portion. When the volume of each part was examined according to the CT slice level, from high to low, the maximum volume of the squamosal portion was at the slice level of the epitympanic cavity. The volume of the petrosal portion was prominent at the slice level of the epitympanic cavity and the upper antrum. These results that the petrosal portion plays a more important role than the squamosal portion in pneumatization of the temporal bone. This analysis with Target Imaging CT provides useful information for understanding the middle ear pathology resulting from poor pneumatization of the temporal bone. PMID- 7472766 TI - [Revision endoscopic sinus surgery--long-term follow up and operative findings]. AB - Among 170 cases (301 sides) with chronic sinusitis who underwent endoscopic sinus surgery, 15 cases (25 sides) were reoperated on because of unsatisfactory results. We examined 12 such cases, periodically, with an endoscope following the first operation. Postoperative endoscopic management, including removal of polyps in the middle nasal meatus and irrigation of the maxillary sinus with sterilized saline water through the enlarged opening, had been done if necessary. However, stenosis of the middle nasal meatus had gradually progressed and mucous or purulent discharge had increased despite postoperative endoscopic treatment. The middle nasal meatus in 17 of the 25 sides had been closed by polyps or/and adhesion at the time of reoperation. The adhesion occurred between the lateral wall of the anterior ethmoid cavity and the middle turbinate. As a result, the opening of the frontal sinus became narrow or even closed, and pus retention was noted in 14 frontal sinuses during revision surgery. In contrast, only seven maxillary sinuses had pus retention. We concluded that adhesion in the middle nasal meatus holds the key to control of inflammatory changes in the paranasal sinuses. PMID- 7472767 TI - [Electric gustatory threshold in diabetics and its clinical significance]. AB - Electrogustometry was performed in diabetics and the correlation between the electric gustatory threshold and clinical factors of the patients (age, disease period and diabetic complications) were evaluated (n = 50). The results obtained were as follows: 1. The electric gustatory threshold rose with aging. The patient age range for further analysis was confined to 50 approximately 69 years old (n = 34) to exclude age dependency. 2. The patients with longer disease histories tended to show a higher electric gustatory threshold, indicating that taste abnormalities in diabetics progress with disease duration. 3. Elevation of the electric gustatory threshold in diabetics was observed before onset or at the earliest stage of the three major complications (diabetic neuropathy, retinopathy and nephropatho), and the threshold rose further with the progression of diabetic complications. Thus, the electric gustatory threshold is a sensitive indicator of diabetic nephropathy and was demonstrated to be a remarkably useful indicator for the prevention of diabetic complications, since it allows detection of the three major diabetic complications at their earliest stages. PMID- 7472768 TI - [Measurement of cochlear blood flow during exploratory tympanotomy in patients suspected of having perilymphatic fistulas]. AB - Since impairment of cochlear blood flow is one of the factors implicated in the pathophysiology of sudden deafness, we measured cochlear blood flow in nine patients suspected of having perilymphatic fistulas. All but one patient had sudden sensorineural hearing loss. The exceptional case had experienced rotatory vertigo immediately after nose-blowing. In the eight patients with sudden deafness, the average hearing level ranged from 30dB to 80dB. Among these patients, there was a first reported case of recurrent inner ear barotrauma due to diving. This case was a 41-year-old male who had a second episode of sudden deafness six years after the first sudden deafness episode affecting the same ear. During exploratory tympanotomy, the tip of a laser-Doppler probe was attached to the promontory near the anterior superior portion of the round window niche. The outer diameter of the probe was 1.7mm and the distance between the exciting and receiving optic fibers was 1.0mm. Pulsatory movement of the laser Doppler output was observed in all 9 patients even after the middle ear mucosa had been removed. We attempted to measure the effects of carbon dioxide on cochlear blood flow, using methods such as inhalation of carbogen and cessation of respiration. In most cases, however, no significant change in cochlear blood flow was observed with these procedures. PMID- 7472769 TI - [Effects of electrical promontory stimulation and band-noise masker in the suppression of tinnitus]. AB - The effects of electrical promontory stimulation (EPS) and band-noise masker (BNM) were examined in 102 patients suffering from tinnitus. The effects of EPS were as follows: complete suppression in 8 patients (8%), partial suppression in 53 patients (52%), no effect in 40 patients (39%), and exacerbation in 1 patient (1%). The effects of BNM were also studied in the same population of patients and the results were as follows: complete suppression in 8 patients (8%), partial suppression in 39 patients (38%) and no effect in 55 patients (54%). The change in the loudness of tinnitus was objectively evaluated, using the loudness-balance test (LBT), after EPS or BNM. The results of LBT correlated with the subjective complaints of tinnitus. In contrast, the pitch match test (PMT) failed to show any remarkable change after EPS or BNM. EPS was more effective in patients with sudden deafness than in those with presbycusis. On the other hand, BNM was more effective in the patients with presbycusis. The mechanisms of suppression of tinnitus by EPS and BNM are discussed. PMID- 7472770 TI - [Electrocochleographic analysis of the suppression of tinnitus by electrical promontory stimulation and changes in compound action potentials]. AB - We performed Electrical Promontory Stimulation (EPS) for the suppression of tinnitus in 52 patients and measured Compound Action Potentials (CAP) using electrocochleography before and after EPS. We examined the relation between residual inhibition and CAP. The purpose of this study was to investigate the origin and mechanism of suppression of tinnitus. Tinnitus was suppressed in 27 patients (52%) after EPS. In the group of patients in which we suppressed tinnitus, the CAP amplitudes increased from 3.0 +/- 1.5 microV to 3.2 +/- 1.6 microV (p < 0.05), the loudness of tinnitus was reduced from 7.3 +/- 4.7 dBSL to 5.6 +/- 5.1 dBSL (p < 0.05) and the CAP latencies did not exhibit remarkable change. In the group of patients in which we did not suppress tinnitus, the CAP amplitudes, latencies and the loudness of tinnitus did not change remarkably. These data indicate that EPS affects auditory nerve fibers directly. It was reported that EPS is effective for tinnitus of peripheral origin and not for that of central origin and, that in tinnitus of peripheral origin, the spontaneous discharges of the auditory nerve increase in number. It is hypothesized that EPS affects auditory nerve fibers and suppresses tinnitus by synchronizing these spontaneous discharges. Side effects of EPS were cough and discomfort of the throat, nose, lips and oral cavity. These effects were observed only during electrical stimulation and there were no permanent side effects. In conclusion, EPS affects auditory nerve fibers directly, is effective for tinnitus of peripheral origin and is not associated with serious complications. PMID- 7472771 TI - [An analytical study on recognition confusion of vowels in patients with 22 channel cochlear implant]. AB - Vowel confusion was studied in 16 postlingually deaf patients with 22 multi channel cochlear implants. Tests for vowel perception were performed on these patients after a 3 month training period, using/a,e,i,o,u/pronounced by a speech therapist. Analyses were conducted on the relationship between the vowel confusion matrix and the frequencies of the 1st and 2nd formants of vowels used in this study, and the relationship between the confusion matrix and the numbers in order of the activating electrodes. The results revealed that the higher percentages of errors in vowel recognition were as follows: /o/-->/a/ (12%),/o/- >/u/ (13%), and/e/-->/i/ (8%), possibly due to small differences found in the F1 and F2 frequency ratio or in the F2 frequency of the vowels, which results in a similar distance between two electrodes, pairs or similar stimulated places in the sense of the high frequency part of the cochlea. PMID- 7472772 TI - Retention of CD44 introns in bladder cancer: understanding the alternative splicing of pre-mRNA opens new insights into the pathogenesis of human cancers. PMID- 7472773 TI - Fractal patterns for cells in culture. AB - The present study aims to understand the growth of human malignant tumours in vitro, using the geometry of fractals as a method of analysis. The fractal dimensions of HN-5 and MDCK cell growth patterns have been measured. The first results may suggest the possibility of distinct growth processes characterized by different (time-dependent) effective fractal dimensions for MDCK and HN-5 cells. If this is true, the fractal dimensions may yet prove to be a useful discriminant for comparing different diagnostic categories. PMID- 7472774 TI - Unusual retention of introns in CD44 gene transcripts in bladder cancer provides new diagnostic and clinical oncological opportunities. AB - Increased and disorganised expression of CD44 variant exons has been demonstrated in biopsy samples of several types of human malignancy by many groups. This abnormality can be used to detect exfoliated tumour cells in the urine of bladder cancer patients. The present report demonstrates that the deranged activity of this gene in neoplasia also results in the accumulation of immature mRNA transcripts containing non-coding sequences (introns) from this gene in cancer tissues and cells. There is simultaneous overexpression of a newly identified 437 bp exon (coding sequence) located in the variably active region of the gene and of many abnormal variant exon combinations. This is the first report describing the specific retention of introns in gene transcripts in clinical diagnostic samples of tumour tissues and cells. The phenomenon was seen repeatedly in samples from cancer patients and in a cancer cell line, and thus could form the basis for a unique new and specific method of cancer diagnosis. PMID- 7472775 TI - Analysis of the BCL-6 gene configuration in diffuse B-cell non-Hodgkin's lymphomas and Hodgkin's disease. AB - BCL-6 is a novel proto-oncogene that codes for a zinc-finger protein sharing homologies with many transcription factors. It has recently been shown that BCL-6 is involved in chromosome band 3q27 aberrations in non-Hodgkin's lymphomas (NHLs) and BCL-6 rearrangements have been detected in 34-45 per cent of diffuse large cell lymphomas with B immunophenotype. We have studied the BCL-6 gene configuration by Southern blot analysis in 60 cases of B-cell NHL and in 17 cases of Hodgkin's disease (HD). BCL-6 was rearranged in 15/46 (32.6 per cent) diffuse B-large cell lymphomas, mainly with centroblastic morphology, and in 2/11 (18.2 per cent) follicular (centroblastic-centrocytic) lymphomas. Conversely, all cases of HD, including four cases of lymphocyte predominant, nodular type (nodular paragranuloma), had a germline configuration. These findings confirm that BCL-6 is rearranged in a significant percentage of diffuse B-large cell lymphomas, suggesting that this proto-oncogene might have a pathogenetic role in this subset of NHLs, but our preliminary analysis suggests that BCL-6 lesions are not involved in the pathogenesis of HD. However, further investigations using more sensitive techniques are required to confirm these findings. PMID- 7472776 TI - MDM2 expression in lymphoid cells and reactive and neoplastic lymphoid tissue. Comparative study with p53 expression. AB - MDM2 and p53 immunohistochemical protein expression was analysed in lymphocytes and in reactive and neoplastic lymphoid tissue. Phytohaemagglutinin (PHA) stimulated lymphocytes displayed MDM2 and p53 co-expression. In 8 of 8 tonsils, 24 of 24 Hodgkin's disease (HD), and 10 of 24 high-grade non-Hodgkin's lymphoma (HG-NHL) specimens, MDM2 paralleled p53 nuclear expression in non-tumour and tumour cells. The number of positive cells was greater and the staining intensity was stronger for p53 than for MDM2. In another nine of the 24 HG-NHL cases studied, dissociated expression was observed, with high p53 expression and very low or absent MDM2 expression. In five cases, both MDM2 and p53 were negative. The eight low-grade NHL (LG-NHL) cases were also MDM2- and p53-negative. MDM2 and p53 expression in PHA-activated lymphocytes and reactive lymphoid tissue is probably an expression of opposing biological signals regulating cell proliferation. Parallel MDM2 and p53 expression in all HD and in 10 out of 24 HG/NHL cases may indicate that this growth suppressive pathway is maintained in those cases. However, dissociated MDM2/p53 expression (nine cases) and the absence of expression of both proteins (five cases) may represent examples of deregulation of this growth control pathway. These findings are in agreement with previous in vitro studies in cell lines regarding the role of MDM2/p53 lymphoid tissue, suggesting a possible role for MDM2 deregulation in lymphomagenesis. PMID- 7472777 TI - High frequency of light chain restriction in labial gland biopsies of Sjogren's syndrome detected by in situ hybridization. AB - A well-recognized complication of Sjogren's syndrome (SS) is the development of malignant lymphoma, with a risk 44 times that of the general population. Although a few clinical signs may indicate the onset of lymphoma, there are few reliable laboratory markers which predict the development of neoplasia. A non-isotopic in situ hybridization technique has been applied to routinely processed labial salivary gland (LSG) biopsies of patients under investigation for SS. Serial section of 70 LSGs were examined for a kappa and gamma immunoglobulin light chain mRNA using digoxigenin-labelled oligonucleotide probes. As controls, 39 biopsies from non-SS-associated sialadenitis were also examined. Sections were analysed using computer-assisted quantification to determine the percentage of kappa expressing cells in each case. The range of kappa expression in the SS group was 24.1-93.4 percent and in the non-SS group 48.3-75.4 per cent. Light chain restriction was found in 13/70 (18.6 percent) cases from the SS group but in no cases of the control group. Of the SS cases showing restriction, 4/13 (30.7 percent) have subsequently developed extrasalivary gland lymphoma. Two patients not showing light chain restriction in LSG have subsequently developed lymphoma. The positive predictive value of this test to identify patients at risk of lymphoma was 30.7 percent with a detection rate (sensitivity) of 66.7 percent and a false-positive rate of 14.1 per cent(specificity 85.9 percent). This study has identified a high prevalence of light chain restriction in labial gland biopsies of patients with SS and provides objective quantitative criteria to identify those patients at greater risk of lymphoma development. PMID- 7472778 TI - CD25-negative hairy cell leukaemia: intracytoplasmic detection of Tac antigen and interferon-induced surface expression. AB - Hairy cell leukaemia (HCL) is a chronic lymphoproliferative disease of B-cell lineage. One of the peculiar immunophenotypic markers is the strong expression of the p55 chain of the interleukin-2 receptor (IL2R), recognized by anti-CD25 (or anti-Tac) monoclonal antibody. However, it is known that in rare cases CD25 may not be detectable, even when variant forms of HCL are excluded. The possibility has not been investigated that in these situations CD25 is present in the cytoplasm of the neoplastic cells. This paper describes a case in which the clinical, histological, and electron microscopic features were consistent with a typical HCL. Immunophenotype analysis showed the whole spectrum of markers of HCL, except for the expression of IL2R. The soluble form of the molecule was, however, increased in the patient's serum. Cytospin staining of the neoplastic B cells with anti-CD25 clearly demonstrated the presence of IL2R in the cytoplasm of hairy cells. When the cells were cultivated in vitro in the presence of interferon-alpha 2b, CD25 was detectable at the membrane level. These findings suggest that at least some cases of CD25-negative HCL may express cytoplasmic IL2R. PMID- 7472779 TI - Apoptosis suppressing protein bcl-2 is expressed in well-differentiated breast carcinomas with favourable prognosis. AB - The expression of bcl-2 protein was analysed immunohistochemically in 202 female breast carcinomas. The intensity of bcl-2 expression was inversely related to tumour grade (P < 0.0001), tumor necrosis (P < 0.0001), mitotic index (P < 0.0001), oestrogen receptor content (P < 0.0001), progesterone receptor content (P = 0.0007), S-phase fraction (P = 0.00047), and apoptotic index (P = 0.087). A high fraction of bcl-2-positive cells was related to ductal or lobular type (P = 0.03) and slight nuclear pleomorphism (P = 0.03). Heterogeneous expression of bcl 2 protein was associated with high grade (P = 0.02), severe nuclear pleomorphism (P = 0.02), DNA aneuploidy (P = 0.018), high S-phase fraction (P = 0.05), and early metastasis (P = 0.03). Intense expression of bcl-2 protein was significantly related to favourable outcome in the entire cohort (P = 0.0013), as well as in axillary lymph node-negative (ANN) tumours (P = 0.0124). Long recurrence-free periods in the entire cohort (P = 0.037) and in ANN tumours (P = 0.08) were confined to cases with intense expression of bcl-2 protein. In multivariate analysis, bcl-2 expression had no independent prognostic value in the entire cohort or in axillary lymph node-negative breast carcinomas, whereas it was a weak independent prognostic factor in axillary lymph node-positive breast carcinomas. PMID- 7472780 TI - Funding the clinical autopsy. PMID- 7472781 TI - Microvessel count predicts metastatic disease and survival in non-small cell lung cancer. AB - The growth of newly formed vessels, or neoangiogenesis, represents an important step in both physiological and pathological situations: in particular, tumour growth and metastasis require angiogenesis. Microvessel count (MC), which represents a measure of tumour angiogenesis, has been associated with metastatic spread in cutaneous, mammary, prostatic, head and neck, and early-stage lung cancer. In this study, the role of tumour angiogenesis as a prognostic indicator was examined in 253 primary non-small lung cancer (NSCLC) patients. Microvessels were counted by highlighting endothelial cells with anti-Factor VIII monoclonal antibody (Mab) in methacarn-fixed tumour samples. In univariat analysis, MC (P< 0.000001), sex (P=0.0036), histotype (P < 0.014), tumour status (P <0.007), and vessel invasion (P < 0.019) were significantly related to hilar and/or mediastinal nodal involvement. However, in the stepwise logistic regression analysis, MC (P<0.000003) retained the most important influence on nodal metastasis. The overall survival analysis calculated by the Kaplan-Meier method revealed that tumours with high MC ( > 25 vessels/field) were significantly associated with increased death risk (log-rank test P = 0.00067; Cox's test P = 0.00046; Gehan's Wilcoxon test P = 0.00108). In 94 patients, the development of metastatic disease during follow-up was significantly related to MC. Indeed, patients who developed metastasis during follow-up showed a higher MC, either as a dichotomous (P = 0.01) or as a continuous (P = 0.003) variable, than patients who had developed no metastasis at the time of the analysis. Moreover, in the stepwise logistic regression analysis, MC retained the most important influence on distant metastases.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7472782 TI - p53 immunoreactivity and mutation of the p53 gene in smooth muscle tumours of the uterine corpus. AB - Mutation and overexpression of p53 have been described in uterine malignant mixed Mullerian tumours and in endometrial adenocarcinoma, where it has been associated with a poor prognosis. This study examines p53 expression and mutation of the p53 gene in benign and malignant smooth muscle tumours of the uterine corpus. p53 expression was evaluated by immunohistochemistry in formalin-fixed, paraffin embedded tissue from 23 leiomyosarcomas, 10 tumours of uncertain malignant potential (TUMPs), and 18 leiomyomas. Single-stranded conformational polymorphism, (SSCP) analysis of exons 5-8 of the p53 gene was performed on 13 leiomyosarcomas, nine TUMPs, and eight leiomyomas. With microwave antigen retrieval, p53 immunoreactivity was seen in 13/23 microwave treatment, staining was abolished in three leiomyosarcomas, all immunoreactive TUMPs, and the single positive leiomyoma. SSCP analysis revealed mutation in three leiomyosarcomas. There was one mutation in exon 5 in a case with positive immunohistochemistry. Two cases with negative staining showed mutation, one in exon 7 and one in exon 8. Mutation was present in exon 7 in 4/9 and in exon 6 in 1/9 TUMPs. All of these cases showed positive immunohistochemistry. There was no significant difference in outcome between cases with and without positive immunohistochemistry. p53 expression is seen in a significant proportion of uterine leiomyosarcomas. Microwave antigen retrieval increases the proportion of positive cases and also results in positive staining in TUMPs. Mutation of the p53 gene occurs in only a minority of leiomyosarcomas and in a significant proportion of TUMPs. Positive immunohistochemistry does not, however, correlate with the presence of mutation and other factors may be responsible for p53 detection in many cases. PMID- 7472783 TI - Ultrastructure of rat pulmonary arterioles after neonatal exposure to hypoxia and subsequent relief and treatment with monocrotaline. AB - A group of rats was born in and spent the first 4 weeks of life at a simulated altitude of 3550 m. Two animals were killed immediately afterwards and the remaining 16 were allowed to recover for various times up to a maximum of 12 weeks at sea-level atmospheric pressure. On ultrastructural examination, the pulmonary arterioles of hypoxic rats showed muscularization, the new layer of mature smooth muscle cells containing abundant organelles and myofilaments. These cells were bounded by prominent elastic laminae. During the recovery period, the medial layer became progressively thinned, but the cells still retained some characteristics of smooth muscle by 12 weeks' recovery. When a similar group of ten hypoxic rats was allowed to recover for 12 weeks before being given monocrotaline, there was early enlargement of the residual smooth muscle cells in the media of pulmonary arterioles and within 5 weeks there was again a thick layer of medial smooth muscle. This was in contrast to the sparse, weakly muscularized arterioles seen in eight similarly treated rats born under normoxic conditions. The relevance is discussed of these findings to the rare occurrence of primary pulmonary hypertension in people who were born at high altitude but returned to sea-level during childhood. PMID- 7472784 TI - The hypotransferrinaemic mouse: ultrastructural and laser microprobe analysis observations. AB - Homozygote hypotransferrinaemic mice (hpx/hpx) have cytopathological features similar to those of human congenital atransferrinaemia, genetic haemochromatosis, and neonatal haemochromatosis. These conditions all have in common high levels of cytotoxic non-transferrin-bound serum iron. This study describes the ultrastructural features of iron overload in liver, pancreas, heart, and small intestine of 2- and 12-month-old hypotransferrinaemic mice. Electron microscopic studies of unstained sections showed early parenchymal cell siderosis, with accumulation of numerous ferritin particles and clusters in the cytosol, as well as ferritin and haemosiderin in lysosomes (siderosomes). In the 12-month-old animals, iron was also found in Kupffer cells and macrophages in other tissues. In addition, there were conspicuous iron-containing compounds in the bile canaliculi, and marked iron deposition in the pancreas and heart. Laser microprobe mass analysis (LAMMA) enabled localization and relative quantitation of iron deposition in subcellular compartments providing in situ documentation of iron accumulation in siderosomes and contributed in assessing total cytosolic iron in various cell types. Moreover, it demonstrated the importance and magnitude of the biliary route for iron excretion in these animals. PMID- 7472785 TI - An infarct-like myocardial lesion experimentally induced in Wistar rats with Africanized bee venom. AB - There is evidence in the literature that death following a bee or wasp sting may result from cardiac involvement. This study describes acute cardiac lesions experimentally induced in Wistar Rats submitted to intravenous inoculation of Africanized bee venom (ABV) and killed 1, 4, and 24 h after inoculation. Significant increases in serum enzyme levels were detected; light microscopy showed necrosis of the myocardium; and enzyme histochemistry showed inactivation of enzymes in and around the areas of necrosis. This is the first report of an acute necrotizing cardiac lesion, similar to human myocardial infarction, produced by the inoculation of ABV. PMID- 7472787 TI - Factors associated with length of survival among 1081 terminally ill cancer patients. AB - To improve their ability to estimate the survival of terminally ill cancer patients, palliative care physicians require accurate information on prognostic factors. The objective of this study was to assess the extent to which variables such as patient characteristics and primary tumor site affect the length of survival of terminally ill cancer patients. The study population consisted of 1081 cancer patients admitted for terminal care to a 15-bed palliative care unit from 1985 to 1991. Univariate Kaplan-Meier survival analysis and multivariate Cox regression analyses were used to examine the relationship between patient characteristics at admission and survival time. The factor most strongly associated with shorter survival was poor performance status; this strong relationship was not altered by taking into account sex and primary cancer site in the multivariate analysis. For patients who were bedridden at admission, the death rate was 5.5 times higher (95% confidence interval (Cl) 3.4-9.0) than that for ambulatory patients during the first four days of stay, and it was 2.8 times higher (95% Cl 2.0-3.9) subsequently (up to 19 days). The other prognostic factors significantly but slightly associated with poorer survival in the univariate analysis were primary lung cancer, male sex, and living with a spouse. These findings indicate that performance status is the main prognostic factor for accurately estimating the survival time of terminally ill cancer patients. PMID- 7472788 TI - Dying of non-malignant diseases in general practice. AB - Recent policy changes within the British National Health Service have encouraged the provision of care within the community. The objective of this study was to explore the needs of informal caregivers of people with non-malignant diseases who wished to remain in their own homes until death. Semi-structured interviews with a non-random sample of 20 bereaved carers from 14 surgeries, their general practitioners, and district nurses were analyzed to discover the extent and quality of the care provided by the primary care team. One quarter of the doctors considered that palliative care applied exclusively to people with cancer. One half of the doctors felt that palliative care was primarily concerned with pain relief. Both doctors and nurses believed that their role was that of coordinating care for the patient. Carers were reluctant to criticize professional caregivers, but the results of this study suggest that carers need help with the burden of caring, including emotional support, advice and information regarding social services and financial benefits, and recognition of their caring role. Palliative care is not exclusive to people with cancer but should be extended to people with non-malignant diseases, encompassing not only symptom relief but also explicitly including emotional or spiritual support, care of the family, and support in preparing for the end of life. PMID- 7472786 TI - What do palliative staff think about audit? AB - Forty-one of 68 palliative care clinicians responded to an anonymous questionnaire assessing the attitudinal, training, management, and support needs for introducing audit. Most respondents (88%) felt it was a good idea; reservations concerned the increase in work load. Although 66% of respondents had received audit training, only 39% were currently involved in audit. Respondents identified a need for more information and feedback, experienced assistance, and support. These concerns have to be addressed by health care managers if audit is to be successfully maintained. Current training sessions do not adequately meet staff needs; palliative care staff require an advanced audit training that includes staff motivation, analysis of results, and the provision of feedback. PMID- 7472789 TI - Is humanity too costly a place to keep open? PMID- 7472790 TI - Evaluation of bereavement anniversary cards. AB - The anniversary of a death is perceived as a difficult time by the majority of closely bereaved relatives and friends. Bereavement anniversary cards are supportive and useful in the context of wider follow-up. This study identified no strong predictors to indicate those for whom such a card would be inappropriate. Several recipients thought that the card solicited financial support for the hospice. PMID- 7472791 TI - Images of palliative nursing care. PMID- 7472792 TI - The role of volunteers in a hospital-based palliative care service. PMID- 7472793 TI - Scarborough's Palliative 'At-home' Care Team (PACT): a model for a suburban physician palliative care team. PMID- 7472794 TI - Directions for palliative care nursing in Canada: report of a national survey. AB - This paper reports the results of a national survey of palliative care nurses conducted following a meeting of nurses at the Fifth Canadian Palliative Care Association Conference in 1993. The intent of the survey was to obtain information about Canadian palliative care nurses's perceptions of practice and professional issues. Eighty percent of respondents believed that palliative care nurses should form a palliative care nurses' organization, with the majority recommending that this occur under the auspices of the Canadian Palliative Care Association. Key issues of importance to palliative care nurses were identified. The two major issues of concern were (a) the need to develop standards of practice and (b) educational needs of palliative care nurses. Respondents also emphasized the importance of maintaining and fostering an interdisciplinary approach to palliative care. Results of this survey are to be further discussed at the Sixth Canadian Palliative Care Association Conference to be held in Halifax in October 1995. PMID- 7472796 TI - Informal caregivers of persons with AIDS. PMID- 7472795 TI - Rehearsing one's death. PMID- 7472797 TI - When does palliative care begin? A needs assessment of cancer patients with recurrent disease. PMID- 7472798 TI - Restructuring the principle of medical futility. AB - This essay surveys the need for a clear and objective definition of medical futility. It is urged that once agreement is obtained for structuring operational guidelines for determining futility, a three-tier decisional structure can be developed for testing whether a given treatment falls within the scope of these guidelines. Under the first tier, the treating physician would be given the primary responsibility for making the determination to withhold treatment on the grounds of futility. While the physician would be under a duty not to prescribe treatment deemed futile, he would be obliged to inform the patient and his family of this decision, including the reasons for it, in order to allow, under the second tier, for an appeal to be taken by the patient or family to the hospital ethics committee. The third tier recognizes a right of limited appeal to the courts. PMID- 7472800 TI - Hydrogen bonding in a non-steroidal anti-inflammatory drug--Naproxen. AB - Photophysical properties of a non-steroidal anti-inflammatory drug, Naproxen (6 methoxy alpha-methyl-2-naphthalene acetic acid sodium salt), were investigated in solvents of different polarity, hydrogen donor ability and also in cyclodextrins. The results indicate that in all cases the emitting state is the 1L(b) singlet. In alcoholic solvents, an intermolecular hydrogen bond is responsible for the observed photophysical behaviour of the probe whereas in non-protic solvents (polar and weakly polar) an intramolecular hydrogen bond type is postulated to rationalize the data found. In water, the non-radiative rate constant has a value similar to those found in aqueous solutions of alpha- and beta-cyclodextrins where the probe form complexes. The behaviour in water is explained by a water structure enforced hydrophobic effect. The spectroscopic results are interpreted on the basis of a multiple-parameter model that considers specific solute-solvent interactions. These were also observed in the ground state and detected by Fourier transform infrared spectroscopy. Molecular mechanics (MM) and molecular orbital (AM1) calculations also support the existence of two conformations (rotamers) in Naproxen with non-equivalent intramolecular hydrogen bond-like formation. PMID- 7472799 TI - In vivo spectroscopic properties of the fluorescent pH indicator biscarboxyethyl carboxyfluorescein. AB - Transcutaneous detection of fluorescence from an injection of the pH-sensitive probe biscarboxyethyl carboxyfluorescein (BCECF) has been used to monitor plasma pH in conscious animals. The fluorescence signal must be calibrated with reference to a standard curve. This standard, calibration curve has been achieved using in vitro methods. Here it is shown that temperature influences the calibration curve determination in vitro and hence influences the pH determined from transcutaneous measurements. Two calibration curves have been obtained, one at room temperature (approximately 25 degrees C) and the other at 37 degrees C. At pH 7.01 the calibration curves intersect, so that, at more alkaline pH values, use of room temperature calibration data will lead to an overestimate of plasma pH. Below pH 7.01, plasma pH will be underestimated. Transcutaneous fluorescence spectra recorded from a mouse injected with BCECF are shown, indicating that baseline plasma pH was estimated about 0.3 pH units too high using room temperature calibration, and that the extent of acidification when the animal was allowed to breathe an atmosphere containing 15% CO2 was overestimated by 0.7 pH units. Additionally, it is shown that in vitro bovine serum albumin at concentrations comparable with albumin concentrations in vivo shifts the absorption, fluorescence excitation and emission spectra of BCECF. However, fluorescence spectra recorded in vivo show no such shift. The results indicate that in vitro calibration for transcutaneous fluorescence measurements in vivo can be misleading, and that in the case of pH measurement inattention to temperature can lead to spurious results. PMID- 7472801 TI - Visible light induced changes in the immune response through an eye-brain mechanism (photoneuroimmunology). AB - The immune system is susceptible to a variety of stresses. Recent work in neuroimmunology has begun to define how mood alteration, stress, the seasons, and daily rhythms can have a profound effect on immune response through hormonal modifications. Central to these factors may be light through an eye-brain hormonal modulation. In adult primates, only visible light (400-700 nm) is received by the retina. This photic energy is then transduced and delivered to the visual cortex and by an alternative pathway to the suprachiasmatic nucleus (SCN). The SCN is a part of the hypothalamic region in the brain believed to direct circadian rhythm. Visible light exposure also modulates the pituitary and pineal gland which leads to neuroendocrine changes. Melatonin, norepinephrine and acetylcholine decrease with light activation, while cortisol, serotonin, gaba and dopamine levels increase. The synthesis of vasoactive intestinal polypeptide (VIP), gastrin releasing peptide (GRP) and neuropeptide Y (NPY) in rat SCN has been shown to be modified by light. These induced neuroendocrine changes can lead to alterations in mood and circadian rhythm. All of these neuroendocrine changes can lead to immune modulation. An alternative pathway for immune modulation by light is through the skin. Visible light (400-700 nm) can penetrate epidermal and dermal layers of the skin and may directly interact with circulating lymphocytes to modulate immune function. However, even in the presence of phototoxic agents such as eosin and rose bengal, visible light did not produce suppression of contact hypersensitivity with suppresser cells. In contrast to visible light, in vivo exposure to UV-B (280-320 nm) and UV-A (320-400 nm) radiation can only alter normal human immune function by a skin mediated response. Each UV subgroup (B, A) induces an immunosuppressive response but by differing mechanisms involving the regulation of differing interleukins and growth factors. Some effects observed in humans are: inhibition of allergic contact dermatitis; inhibition of delayed hypersensitivity to an injected antigen; prolongation of skin-graft survival and induction of a tumor-susceptible state. The following article will review much of the progress in this field and explore possible areas of future research. PMID- 7472804 TI - Open questions in photobiology II. Induction of nicks by UV-A. PMID- 7472802 TI - Hydrolysis of RRR-alpha-tocopheryl acetate (vitamin E acetate) in the skin and its UV protecting activity (an in vivo study with the rat). AB - Vitamin E acetate is often used rather than vitamin E as an ingredient of skin care products and dermatological preparations, because it lacks the free phenolic OH group. However, because of this the acetate as such is biologically inactive. In spite of this intrinsic inactivity, the skin is protected against the harmful effects of sunlight after topical application of vitamin E acetate. Therefore it is supposed that hydrolysis takes place in the skin and that the reaction product, the radical scavenger vitamin E, is responsible for the protection observed. In this in vivo study with the rat, we have investigated the hydrolysis of RRR-alpha-tocopheryl acetate (vitamin E acetate) in the epidermis in relation to UV radiation protection. (As a measure of protection, we used the UV-induced binding of 8-methoxypsoralen to epidermal biomacromolecules.) After a period of 5 h from a single application of vitamin E acetate, hydrolysis into free vitamin E was not observed. No protection was found at this time point, corresponding with the absence of vitamin E. After treatment for 5 days, consisting of one topical application daily, the percentage of acetate present in the stratum corneum which was hydrolysed into free vitamin E was less than 1%, whereas the corresponding value for the viable layer of the epidermis was about 5%. The hydrolysis of vitamin E acetate in the epidermis proceeded very slowly. As a result, the absolute amount of free vitamin E, found in the total epidermis after treatment for 5 days with the acetate, was only a few times higher than the normal level. Yet, this very small amount of free vitamin E proved to be sufficient for maximal protection in this animal model. The results show that vitamin E acetate acts as a prodrug, which very slowly releases minute amounts of active vitamin E. PMID- 7472803 TI - Repetitive photodynamic therapy with topical delta-aminolaevulinic acid as an appropriate approach to the routine treatment of superficial non-melanoma skin tumours. AB - Photodynamic therapy (PDT) with topical delta-aminolaevulinic acid (ALA) is considered as a valuable tool for treating non-melanoma skin tumours, but there is no consensus about the methods of treatment, where they should be used and the rates of complete responses. This study reports the treatment of 50 actinic keratoses (AKs), 23 superficial basal cell carcinomas (BCCs), 30 nodular BCCs, 4 pigmented BCCs, 12 superficial squamous cell carcinomas (SCCs), 6 nodular SCCs, 6 Bowen's diseases and 4 keratoacanthomas (KAs). A 20% ALA emulsion was applied under an occlusive dressing for 6-8 h and the skin was then irradiated with 630 nm light from a dye laser. Treatments were delivered every other day until complete clinical disappearance of the lesions was observed. ALA-PDT was interrupted in the case of a partial response if, after two additional treatments, no further improvement was observed. All AKs (50/50), superficial BCCs (23/23), Bowen's diseases (6/6) and KAs (4/4) showed a complete response. In addition, 91.6% (11/12) superficial SCCs, 80.0% (24/30) nodular BCCs and 66.7% (4/6) nodular SCCs responded completely to the treatment. All 4 pigmented BCCs were resistant to the therapy. Some (59) of the treated areas that appeared completely responsive were excised for serial histopathological examination and the remaining 63 were followed-up for 24-36 months (median value, 29 months). Comprehensively, the rates of complete responses decreased after taking into consideration the results of these histological examinations and the long-term follow-up. The final response rates are as follows: 84.0% (42/50) AKs, 86.9% (20/23) superficial BCCs, 50% (15/30) nodular BCCs, 83.3% (10/12) superficial SCCs and 33.3% (2/6) nodular SCCs. The rates of complete responses of Bowen's diseases and KAs remained at 100%.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7472805 TI - Open questions in photobiology III. Melanin and photoprotection. PMID- 7472806 TI - On the role of protoporphyrin IX photoproducts in photodynamic therapy. PMID- 7472807 TI - Photophysical properties of porphyrins in biological membranes. AB - This review illustrates the photophysical properties of some porphyrins, especially those used for biomedical applications, in relation to their photosensitizing efficiency in biological membranes. Porphyrin absorption and luminescence properties are mainly examined. The factors influencing the affinity of porphyrins for biological membranes, including the dye hydrophobicity, the charge and aggregation state, the pH of the medium and the physicochemical properties of the dye environment, are discussed. These factors determine the differences in the photophysical properties of porphyrins in biological membranes. Particular attention is paid to the porphyrin aggregation state: only monomeric species and possibly planar end-to-end aggregates are endowed with significant photosensitizing ability. Many conclusions presented are based on data obtained on membrane model systems such as micelles or liposomes which can mimic specific situations occurring in cells. PMID- 7472808 TI - The reduction of thymine residues in DNA by the combined action of UV light and hypophosphite. AB - UV irradiation (lambda = 254 nm) of thymine and uracil in aqueous solution containing salts of phosphinic acid (hypophosphites) results in the formation of the corresponding dihydropyrimidine derivatives. The peculiarities of this new photochemical reaction consist of a specificity towards 2,4-dioxophyrimidines, a high quantum yield and a neutral pH optimum. The quantum yield of photoconversion of thymine at pH 7.0 is equal to 1.9 x 10(-2) in 1 M NaH2PO2; it is diminished to 8.5 x 10(-3) in 0.1 M NaH2PO2. The same decrease in quantum yield with concentration is also found for uracil and uridine; quantum yields of their transformations in 0.1 M NaH2PO2 at pH 7.0 are 6.6 x 10(-2) and 1.5 x 10(-1) respectively. The mild conditions and high quantum yields characteristic of the photoinduced reaction above open up the possibility of obtaining DNA molecules which contain among pyrimidine photoproducts mainly dihydropyrimidine residues. A correlation between various types of thymine modifications in UV-irradiated double-stranded DNA (dimers and dihydrothymidine residues) and the amplitude of the intense negative band in the circular dichroism (CD) spectra specific for DNA liquid crystalline dispersions has been established. A possible application of this base-specific modification to the investigation of nucleic acids is considered. PMID- 7472809 TI - Molecular mechanism of drug photosensitization. Part 6. Photohaemolysis sensitized by tolmetin. AB - The photosensitizing properties of tolmetin, 5-(p-toluoyl)-1-methyl-2 pyrrolyacetic acid (TLM), have been studied in vitro following the lysis of erythrocytes in phosphate buffer suspensions irradiated with UVA light in the presence of the drug. It was found that the phototoxic properties of the drug are negligible in nitrogen and significant in aerated medium, but that they decrease in oxygen-saturated solution. The investigation of the drug photolysis showed that TLM undergoes photodecarboxylation to p-tolyl 1,2-dimethyl-5-pyrrolyl ketone in nitrogen and to p-tolyl 1-methyl-2-hydroxymethyl-5-pyrrolyl ketone and 5-(p toluoyl)-1-methyl-2-pyrrole carbaldehyde in air. These photoproducts also undergo photodegradation. The comparison between the photohaemolysis and photolysis results and the effect of suitable additives such as sodium azide, mannitol, butylated hydroxy-anisole, reduced glutathione, superoxide dismutase and copper (II) suggest that the phototoxicity of TLM can be attributed essentially to singlet oxygen in the first step and to its photoproducts when they accumulate and compete with the starting drug in light absorption. Their phototoxic effect is much higher with respect to that of TLM, as shown by comparison of the doses needed to attain 50% photohaemolysis. PMID- 7472810 TI - Role of the centrosome in mitosis: UV micro-irradiation study. AB - Ultraviolet micro-irradiation (UV-MI) of the PK (pig kidney embryo) cell centrosome (lambda max = 280 nm, spot diameter 1.6 mm, exposure time 5-15 s) at metaphase and anaphase resulted in functional damage of the centrosome. After UV MI of the centrosome at early metaphase, chromosomes quickly (in 1-3 min) moved away from the irradiated pole and then encircled the non-irradiated pole. Within 10 min after UV-MI the spindle disassembled and chromosomes remained unseparated. The minimal dose inducing this effect in 90% of cells was accumulated in 5 s. After the same UV-MI at late metaphase, chromosomes shifted towards the non irradiated pole; however, anaphase started and chromosome motion towards the non irradiated pole continued normally. UV-MI of the centrosome at early anaphase for 5-15 s slowed down and then stopped chromosome motion towards the irradiated pole. This was a result of rapid (within 2-3 min) disorganization of the half spindle. Chromosomes continued to move towards the opposite pole normally, while cytokinesis was significantly retarded. No visible lesion was revealed by electron microscopy after 5 s UV-MI, while 15 s irradiation resulted in the truncation of the microtubule bundles 1.5-2 microns from the centrosome. We concluded that UV-MI inactivates the centrosome and induces disaggregation of microtubule initiation sites. The critical point (checkpoint) in mitosis up to which this damage induces mitotic arrest is mid-metaphase. PMID- 7472811 TI - Study of porphyrin fluorescence in tissue samples of tumour-bearing mice. AB - A time-gated fluorescence-imaging technique was applied to study the distribution of sensitizer in porphyrin-treated tumour-bearing mice. The animals were administered with either haematoporphyrin derivative (HpD) or Photofrin and sacrificed 4 or 12 h later. Fluorescence images were acquired from tumour, skin, muscle, fat, brain, lymph node, bowel and bone of both treated and untreated mice. The results obtained with HpD and Photofrin are similar. In images acquired 30 ns after excitation a bright exogenous fluorescence allows clear detection of the tumour. Nevertheless, the images show that porphyrins localize with different concentrations in all the examined tissues except the brain. Moreover, an appreciable long-living endogenous emission was observed in the bone. PMID- 7472812 TI - Effect of haematoporphyrin-induced photosensitization on lipid membranes. AB - In vitro cultured mouse myeloma (Sp-2/0-Ag14) cells and phosphatidylcholine liposomes were used to study the membrane effects of photosensitization with an He-Ne laser activated haematoporphyrin (HP). Lipophilic HP molecules, intercalated between the membrane lipid molecules, caused morphological changes of cell membranes on light activation. Steady state and time-resolved fluorescence spectroscopic studies of membrane-bound HP molecules provide information about the change in membrane lipid dynamics (fluidity). Increased HP fluorescence anisotropy was found after laser irradiation in the case of cell membrane. This finding can be related either to the increased rotation correlation time of the rotating fluorophore (HP) (decreased membrane fluidity) or to the decrease in the angular range of molecular rotation, which corresponds to an increased lipid order after photosensitization. Changes in the ratio of saturated:unsaturated fatty acid content of membrane lipids or other chemical events such as cross-linking of membrane components during the photosensitization process can also account for the observed effects. PMID- 7472813 TI - Treatment with 8-MOP and UVA enhances MHC class I synthesis in RMA cells: preliminary results. AB - The response of psoriasis and cutaneous T-cell lymphoma to treatment with 8 methoxypsoralen (8-MOP) and long wavelength ultraviolet light (UVA) is only partly understood. Psoralens form photoadducts within the DNA after activation by UVA and this damage leads to the inhibition of DNA synthesis. Additionally, it has been shown that different forms of DNA damage can induce a stress response, leading to upregulation of selected products. Among these are the major histocompatibility complex (MHC) class I genes. Thus the aim of the present study was to assess the rate of synthesis of MHC class I proteins in murine T-cell lymphoma cells (RMA) after treatment with 8-MOP and UVA. RMA cells were treated with 8-MOP (50-200 ng ml-1) and UVA (1 J.cm-2) and metabolically labelled with 35S-methionine 4 and 24 h after treatment. MHC class I synthesis was determined by immunoprecipitation of the cell lysates with an anti-Kb monoclonal antibody, Y3. After 4 h, treated and untreated cells demonstrated no differences in the rate of MHC class I synthesis. However, after 24 h a dose-dependent increase in MHC class I synthesis was observed. This increase in MHC class I expression could be responsible, at least partly, for the responses observed in patients treated with photopheresis. PMID- 7472814 TI - A recipe for the preparation of a rodent food that eliminates chlorophyll-based tissue fluorescence. PMID- 7472815 TI - Twin-to-twin transfusion syndrome: new perspectives. PMID- 7472816 TI - Use of pancreatic enzyme supplements for patients with cystic fibrosis in the context of fibrosing colonopathy. Consensus Committee. PMID- 7472817 TI - Discontinuing penicillin prophylaxis in children with sickle cell anemia. Prophylactic Penicillin Study II. AB - OBJECTIVE: To evaluate the consequences of discontinuing penicillin prophylaxis at 5 years of age in children with sickle cell anemia who had received prophylactic penicillin for much of their lives. DESIGN: Randomized, double blind, placebo-controlled trial. SETTING: Eighteen teaching hospitals throughout the United States. PATIENTS: Children with sickle cell anemia (hemoglobin SS or hemoglobin S beta 0-thalassemia) who had received prophylactic penicillin therapy for at least 2 years immediately before their fifth birthday and had received the 23-valent pneumococcal vaccine between 2 and 3 years of age and again at the time of randomization. Of 599 potential candidates, 400 were randomly selected and followed for an average of 3.2 years. INTERVENTIONS: After randomization, patients received the study medication twice daily--either penicillin V potassium, 250 mg, or an identical placebo tablet. Patients were either seen in the clinic or contacted every 3 months thereafter for an interval history and dispensing of the study drug. A physical examination was scheduled every 6 months. MAIN OUTCOME MEASURES: The primary end point was a comparison of the incidence of bacteremia or meningitis caused by Streptococcus pneumoniae in children continuing penicillin prophylaxis versus those receiving the placebo. RESULTS: Six children had a systemic infection caused by S. pneumoniae, four in the placebo group (2.0%; 95% confidence interval 0.5%, 5.0%) and two in the continued penicillin prophylaxis group (1.0%; 95% confidence interval 0.1%, 3.6%) with a relative risk of 0.5 (95% confidence interval 0.1, 2.7). All invasive isolates were either serotype 6(A or B) or serotype 23F. Four of the isolates were penicillin susceptible, and two (one from each treatment group) were penicillin and multiply antibiotic resistant. Adverse effects of the study drug were reported for three patients (nausea, vomiting, or both), one of whom was in the placebo group. CONCLUSION: Children with sickle cell anemia who have not had a prior severe pneumococcal infection or a splenectomy and are receiving comprehensive care may safely stop prophylactic penicillin therapy at 5 years of age. Parents must be aggressively counseled to seek medical attention for all febrile events in children with sickle cell anemia. PMID- 7472818 TI - Deficient hypoxia awakening response in infants of smoking mothers: possible relationship to sudden infant death syndrome. AB - OBJECTIVE: To determine whether fetal exposure to cigarette smoke impairs postnatal chemoreceptor control of ventilation and hypercapnic and hypoxic awakening responses. STUDY DESIGN: Smoking (n = 13) and nonsmoking (n = 34) women were recruited during pregnancy. Serum cotinine levels were measured to assess level of nicotine exposure. The infants were studied at 2 to 3 months of age for ventilatory and awakening responses to hypoxia (17%, 15% and 13% inspired O2) and hypercapnia 4%, 6%, and 8% inspired CO2). Continuous measures were analyzed with unpaired t tests and analysis of variance for repeated measures. Proportions of awakening and periodic breathing were analyzed by means of a comparison of proportions. RESULTS: The infants of smokers had lower birth weights (3022 +/- 566 vs 3518 +/- 491 gm; p < or = 0.005) and were older at the time of study (10.4 +/- 2.8 vs 8.7 +/- 1.3 weeks; p < 0.01) than the control infants. Maternal cotinine levels were higher in smokers (97.8 +/- 107 ng/ml vs no cotinine; p < 0.0001). More infants of smokers failed to awaken with hypoxia than did control infants (54% vs 15%; p = 0.006). The ventilatory responses to hypoxia and hypercapnia were similar in the two groups. All infants awakened with hypercapnia, and there was no difference in the awakening threshold for carbon dioxide (50.3 +/- 4.5 vs 48.3 +/- 5.4 mm Hg; p = 0.28). CONCLUSION: Infants of mothers who smoked during pregnancy have deficient hypoxic awakening responses, which may contribute to the increased risk of sudden infant death syndrome in infants of smoking mothers. PMID- 7472819 TI - Idiopathic steatohepatitis in childhood: a multicenter retrospective study. AB - OBJECTIVE: To describe the clinical, laboratory, and histopathologic features of idiopathic steatohepatitis in children. STUDY DESIGN: Retrospective review of all liver biopsies performed at Boston Children's Hospital, Massachusetts General Hospital, and the University of Massachusetts Medical Center from 1991 to 1994. Chart review was performed when biopsies demonstrated steatosis. RESULTS: Eighty two patients had biopsy-proven hepatic steatosis. Fourteen patients had fatty liver without evidence of inherited, infectious, autoimmune, endocrinologic, toxicologic, or iatrogenic causes. All 14 patients were obese, averaging 159% of ideal body weight (range, 121% to 222%). Nine patients initially had transient abdominal pain, two had hepatomegaly, and one was identified by incidental laboratory evaluation. These 12 patients had biopsies because of persistent elevations of aminotransferase levels. Two other patients without risk factors for steatosis were identified at staging laparotomy for Hodgkin lymphoma. The 10 boys and 4 girls had an average age of 13.5 years (range, 10 to 18 years). Aminotransferase elevations were modest, with aspartate aminotransferase and alanine aminotransferase values averaging 77 +/- 38 IU and 129 +/- 73 IU, respectively. All had imaging studies demonstrating diffuse fatty change. Histologic examination of biopsy specimens revealed varying degrees of steatosis with inflammation and fibrosis. CONCLUSION: Idiopathic steatohepatitis occurs predominantly or exclusively in obese peripubertal children. This entity represents a frequent reason for liver biopsy in this age group. The degree of steatosis, fibrosis, and inflammation does not correlate with symptoms or signs, and significant liver injury with bridging fibrosis may be present. PMID- 7472822 TI - Optic pathway tumors in children: the effect of neurofibromatosis type 1 on clinical manifestations and natural history. AB - OBJECTIVE: To distinguish the clinical manifestations and natural history of optic pathway tumors (OPT) associated with neurofibromatosis type 1 (NF-1 OPT) from that of OPT not associated with NF-1 (non-NF-1 OPT). METHODS: Two groups of children with OPT were compared: (1) 17 children with NF-1 OPT who were followed prospectively, and (2) 19 children with non-NF-1 OPT who were identified retrospectively by a review of medical records. RESULTS: Precocious puberty was a common initial sign in the children with NF-1 OPT (5/17), and was not found in any patients without NF-1. In contrast, children with non-NF-1 OPT had symptoms attributable to increased intracranial pressure (12/19 and nystagmus (5/19); these symptoms were not found in any patient with NF-1. Decreased visual acuity at the time of diagnosis was common in both groups. There was no significant difference between the children with NF-1 OPT and those with non-NF-1 OPT as to age at diagnosis or sex distribution. Optic nerve involvement was more common in NF-1 (p < 0.001). Both isolated and bilateral optic nerve tumors were found exclusively in children with NF-1, whereas chiasmal (p = 0.016) and optic tract involvement (p = 0.001) were more common in those with non-NF-1 OPT. Radiographic evidence of hydrocephalus was found in none of the children with NF-1 OPT compared with 79% of the non-NF-1 OPT group. Progressive disease was seen in 12% of patients with NF-1 OPT compared with 63% of those with non-NF-1 OPT. CONCLUSIONS: Differences exist between NF-1 OPT and non-NF-1 OPT both at the time of diagnosis and during follow-up. Optic pathway tumors caused by NF-1 and non-NF 1 OPT have different biologic properties that distinguish both their initial clinical manifestations and their natural history. PMID- 7472821 TI - Effect of high-affinity anti-Pseudomonas aeruginosa lipopolysaccharide antibodies induced by immunization on the rate of Pseudomonas aeruginosa infection in patients with cystic fibrosis. AB - Patients with cystic fibrosis (CF; N = 26) and with no prior history of infection with Pseudomonas aeruginosa were immunized with an octavalent O-polysaccharide toxin A conjugate vaccine. During the next 4 years, 16 patients (61.5%) remained free of infection and 10 (38.5%) became infected. Total serum antilipopolysaccharide (LPS) antibody levels induced by immunization were comparable in infected and noninfected patients. In contrast, 12 of 16 noninfected versus 3 of 10 infected patients (p = 0.024) mounted and maintained a high-affinity anti-LPS antibody response. When compared retrospectively with the rate in a group of age- and gender-matched, nonimmunized, noncolonized patients with CF, the rate at which P. aeruginosa infections were acquired was significantly lower (p < or = 0.02) among all immunized versus nonimmunized patients during the first 2 years of observation. Subsequently, only those immunized patients who maintained a high-affinity anti-LPS antibody response had a significant reduction (p < or = 0.014) in the rate of infection during years 3 and 4. Smooth, typeable strains of P. aeruginosa predominated among immunized patients; rough, nontypeable strains were most frequently isolated from nonimmunized patients. Mucoid variants were isolated from one immunized patient versus six nonimmunized patients. These results indicate that the induction of a high-affinity P. aeruginosa anti-LPS antibody response can influence the rate of infection in patients with CF. PMID- 7472820 TI - Correlation of sweat chloride concentration with classes of the cystic fibrosis transmembrane conductance regulator gene mutations. AB - OBJECTIVE: To compare differences in epithelial chloride conductance according to class of mutation of the cystic fibrosis transmembrane conductance regulator (CFTR) gene. METHODS: We evaluated the relationship between the functional classes of CFTR mutations and chloride conductance using the first diagnostic sweat chloride concentration in a large cystic fibrosis (CF) population. RESULTS: There was no difference in sweat chloride value value between classes of CFTR mutations that produce no protein (class I), fail to reach the apical membrane because of defective processing (class II), or produce protein that fails to respond to cyclic adenosine monophosphate (class III). Those mutations that produce a cyclic adenosine monophosphate-responsive channel with reduced conductance (class IV) were associated with a significantly lower, intermediate sweat chloride value. However, patients with the mutations that cause reduced synthesis or partially defective processing of normal CFTR (class V) had sweat chloride concentrations similar to those in classes I to III. CONCLUSION: Studies of differences in chloride conductance between functional classes of CFTR mutations provide insight into phenotypic expression of the disease. PMID- 7472823 TI - Neonatal hyperammonemia caused by a defect of carnitine-acylcarnitine translocase. AB - Carnitine-acylcarnitine translocase deficiency is a newly recognized inborn error of metabolism that involves transport of long-chain fatty acids into mitochondria, which in turn impairs mitochondrial beta-oxidation, and ketogenesis. We report a new familial example; the affected twins had neonatal distress, hyperammonemia, and transient intracardiac conduction defects. Clinical and biochemical analysis of both our patients and the two previously reported patients revealed that this inherited defect could be manifested during the neonatal period without any of the signs classically associated with fatty oxidation defects. In contrast, all four patients had sustained and "isolated" hyperammonemia, which could be misinterpreted as being caused by urea cycle defects. We conclude that carnitine-acylcarnitine translocase deficiency is a potential differential diagnosis in neonates with unexplained neonatal hyperammonemia. Cardiac and muscle involvement may represent further early pivotal symptoms. PMID- 7472824 TI - Final height after long-term growth hormone treatment in Turner syndrome. European Study Group. AB - OBJECTIVES: To study final height after long-term growth hormone (GH) treatment in girls with Turner syndrome (TS). PATIENTS: One hundred fifty three patients with TS, participating in five European trials, were included. They started GH treatment in 1987-1989 at an age of 10 years or older. Mean age at start of treatment ranged between 11.7 and 14.6 years among countries and mean bone age between 9.4 and 11.8 years. Fourteen girls were lost to follow-up, leaving 139 for analysis. Most girls have now attained final height (FH), defined as a linear growth velocity (GV) of 4 mm/yr or less, measured over at least 6 months (group 1, n = 56), or near-FH, defined as a GV of 5 to 9 mm/yr (group 2, n = 22). Sixty one girls were still growing 10 mm/yr or more. METHODS AND MAIN RESULTS: At the last measurement, mean (SD) height was 150.7 (4.9) cm in group 1 and 148.5 (5.1) cm in group 2. The differences between FH and projected final height based on extrapolation of the initial height-standard deviation score on Turner syndrome reference values, were 2.9 (3.8) and 3.0 (3.3) cm, respectively. The mean gain over the Bayley-Pinneau prediction of FH was 3.3 (3.9) cm in both groups. No significant differences between countries were found. The range of gains over projected height (-4.7 to 12.1 cm) was large, and 25% of gains were 5 cm or more. Gain over initial projection was strongly related to initial growth delay and to growth response during the first 2 years of treatment. A logistic regression model is presented that predicts gain of more than 5 cm with a positive predictive value of 62% and a negative predictive value of 84%. CONCLUSIONS: Long term GH treatment in girls with TS, starting treatment at a relatively advanced age ( > 10 years) resulted in a modest mean gain in FH of 3 cm, with wide interindividual variation. PMID- 7472825 TI - Duration of hospital stay for apparently healthy newborn infants. PMID- 7472826 TI - Systemic capillary leak syndrome in a child. AB - A child with idiopathic systemic capillary leak syndrome had four episodes between 3 and 3 years 9 months of age, all characterized by the sudden onset of hypovolemic shock without any obvious triggering factor. Between the attacks she is well and free of symptoms. Early establishment of intravenous access facilitates volume replacement during the acute episodes. PMID- 7472827 TI - Neurocognitive deficits in morbidly obese children with obstructive sleep apnea. AB - Neurocognitive abilities were measured in 14 morbidly obese children, five of whom had obstructive sleep apnea as determined by polysomnography. As in adults, children with obstructive sleep apnea had deficits in learning, memory, and vocabulary. Moreover, apneic/hypopneic events were inversely related to memory and learning performance among the entire sample. PMID- 7472828 TI - Prognosis of infantile seborrheic dermatitis. AB - Of 191 children who had had infantile seborrheic dermatitis, 88 were reexamined after 10 years. One child had psoriasis, four had atopic dermatitis, and seven had seborrheic dermatitis, which suggests a link with adult seborrheic dermatitis. A familial tendency toward infantile seborrheic dermatitis was noted, as was an increased incidence of allergy within the family. PMID- 7472829 TI - Cardiac complications in children with Staphylococcus aureus bacteremia. AB - Clinically silent endocarditis was detected in 4 (11%) of 36 hospitalized children with staphylococcal bacteremia who underwent echocardiographic examination. Pericarditis was detected in two further children. Only one child had underlying cardiac disease (patent ductus arteriosus). Echocardiography should be considered in children with staphylococcemia even if an obvious extracardiac focus is apparent. PMID- 7472830 TI - Association of hypertrophic maxillary frenulum with hypoplastic left heart syndrome. AB - We report a previously unrecognized association between a hypertrophic frenulum separating the alveolar portion of the maxillary palatine suture with hypoplastic left heart syndrome in one fetus and seven children. A hypertrophic frenulum was not found in 25 pediatric cardiology patients without hypoplastic left heart syndrome. This finding may provide a marker for suspecting hypoplastic left heart syndrome in the neonate. PMID- 7472831 TI - Demodicidosis in childhood acute lymphoblastic leukemia; an opportunistic infection occurring with immunosuppression. AB - We report demodicidosis in 11 children with acute lymphoblastic leukemia and a mildly pruritic, erythematous papular dermatitis that developed in areas rich in sebaceous glands. Dermodex eruptions were safely and effectively treated with 5% permethrin. Proliferation of commensal parasites of the skin, Dermodex folliculorum and Dermodex brevis may be an opportunistic infection of the skin in the immunocompromised host; the expected abrogation of cell-mediated immunity secondary to lymphocyte depletion predisposes some children given chemotherapy for leukemia to mite proliferation. PMID- 7472832 TI - Leukocyte adhesion deficiency mimicking Hirschsprung disease. AB - An infant had clinical signs suggestive of Hirschsprung disease as the initial manifestation of leukocyte adhesion deficiency. Chromosome studies showed a deletion of the distal third of the long arm of one chromosome 21, and flow cytometric studies confirmed the defective expression of CD18. PMID- 7472833 TI - Association between duration of neonatal hospital stay and readmission rate. AB - OBJECTIVE: A recent rapid decrease in the duration of neonatal hospital stay in Ontario, Canada, enabled us to study the association between healthy infants' age at neonatal hospital discharge and subsequent readmission rates. DESIGN: (1) Population-based and (2) single hospital-based retrospective studies. SETTING AND STUDY POPULATION: (1) A total of 920,554 healthy infants with a birth weight of 2500 gm or more, born in Ontario from 1987 to 1994. (2) Infants with a birth weight of 2500 gm or more, born during the same period, and readmitted before 15 days from home to the Hospital for Sick Children for jaundice or dehydration. MEASUREMENT: Duration of neonatal stay in the hospital and readmission rates were measured, and diagnostic codes were analyzed. Severity of illness was evaluated in infants readmitted to our hospital. RESULTS: In Ontario the mean length of stay decreased from 4.5 days to 2.7 days (p = 0.000), and the readmission rate during the first 2 weeks of life increased from 12.9 to 20.7 per 1000 (p = 0.000). Increased rates of readmission were most marked for jaundice and dehydration. The infants readmitted to our hospital had evidence of increased severity of illness as indicated by higher serum bilirubin and sodium concentrations. Two deaths occurred in infants with hypernatremic dehydration, one in 1992-1993 and another in 1993-1994. CONCLUSIONS: In Ontario, shorter neonatal hospital stay was associated with increased readmission rates for conditions that may not give rise to symptoms or signs on days 1 to 3 of life. In our hospital the severity of jaundice and dehydration in readmitted infants increased. The severity-of-illness data raise the question of whether shorter neonatal hospital stay of apparently healthy infants is always safe. Decisions to discharge infants should be based on rigorous evaluation of individual infants. PMID- 7472834 TI - Mechanisms for episodes of hypoxemia in preterm infants undergoing mechanical ventilation. AB - OBJECTIVE: To ascertain possible mechanisms implicated in the development of transient episodes of hypoxemia (oxygen saturation < 85%) frequently observed in preterm infants undergoing mechanical ventilation, even after the acute phase of respiratory failure has passed. STUDY DESIGN: Tidal flow, airway and esophageal pressure, and oxygen saturation were continuously recorded in 10 infants (mean +/ SD, birth weight 733 +/- 149 gm, gestational age 25.5 +/- 2.2 weeks, age 26.3 +/ 11.9 days) who had repeated episodes of hypoxemia without any evident cause. Measurements of minute ventilation (VE) inspiratory compliance (Ci), and inspiratory resistance (Ri) were compared before and during episodes of hypoxemia. RESULTS: All episodes of hypoxemia were preceded by an active exhalation that produced a mean decrease in end-expiratory lung volume of 6.4 +/- 2.8 ml/kg. The reduction in lung volume was immediately followed by a sudden decrease in tidal flow and volume, despite continuation of mechanical ventilation at the same rate and peak pressure. The resulting hypoventilation was associated with a drop in Ci to approximately one half and an increase in Ri to more than double the baseline values. Approximately 30 seconds after the beginning of hypoventilation, the arterial oxygen saturation reached a hypoxemic level (oxygen saturation < 85%)> CONCLUSION: Most hypoxemic episodes were triggered by an expiratory effort that produced a large decrease in lung volume. This reduction in lung volume probably leads to closure of small airways and the development of intrapulmonary shunts, which would explain the rapid development of hypoxemia. PMID- 7472835 TI - Early echocardiographic prediction of symptomatic patent ductus arteriosus in preterm infants undergoing mechanical ventilation. AB - OBJECTIVE: To identify early echocardiographic markers allowing prediction of subsequent symptomatic patent ductus arteriosus (PDA). METHODS: One hundred sixteen preterm infants ( < 1500 gm) requiring mechanical ventilation underwent echocardiography at a mean postnatal age of 19 hours (range, 7 to 31 hours). Four potential markers were studied: the left atrial to aortic root ratio, pulsed Doppler signal within the course of the duct (ductal diameter), and the direction of postductal aortic diastolic flow. Subsequent ductal closure or significant patency (if suspected clinically) was confirmed echocardiographically. RESULTS: A significant PDA developed in 42 infants (36%). Ductal diameter was the most accurate echocardiographic marker in predicting subsequent significant most accurate echocardiographic marker in predicting subsequent significant PDA. With a ductal diameter of 1.5 mm or greater there were 34 true-positive, 11 false positive, 63 true-negative, and 8 false-negative results, giving a positive likelihood ratio of 5.5 and a negative likelihood ratio of 0.22 for prediction of development of a PDA requiring treatment. The sensitivity was 81% and the specificity was 85%. Only one infant older than 28 weeks of gestational age had a significant PDA, and limiting the analysis to infants younger than 29 weeks of gestation further improved the predictive accuracy of ductal diameter. The positive likelihood ratio was 8.1 and the negative likelihood ratio was 0.19, with a sensitivity of 83% and a specificity of 90%. CONCLUSION: Color Doppler measurement of the internal ductal diameter allows early prediction of significant PDA in preterm infants. PMID- 7472836 TI - Hemodynamic prediction of complications in neonatal respiratory distress syndrome. AB - OBJECTIVES: We hypothesized that the absence of an increase in the Doppler derived aortopulmonary pressure gradient (APPG) across the ductus arteriosus, which reflects pulmonary artery pressure during the first day of life, can predict clinical complications in preterm infants with neonatal respiratory distress syndrome (RDS) STUDY DESIGN: Twenty-nine healthy preterm infants weighing 2210 +/- 244 gm (mean +/- SEM) and 63 infants with RDS weighing 1645 +/- 86 gm were studied with the Doppler ultrasound technique for measurement of the mean APPG at 2, 24, 48, and 72 hours of age. Of infants with RDS, 67% were treated with synthetic surfactant. RESULTS: Spontaneous closure of the ductus arteriosus on the first day of life led to exclusion of 11 infants with RDS and 17 control subjects from the study. Of the remaining 52 infants with RDS, 26 had an increase in the APPG from 2 to 24 hours of age, and APPG values remained low in the other 26 infants. Neonatal complications appeared more frequently (p < 0.001) in distressed infants with little change in APPG values (delta APPG) (22/26) than in infants with high delta APPG (6/26). Six of the infants with high delta APPG values (23%) needed medical or surgical closure of the ductus, but no other acute neonatal complications occurred; 14 (54%) of the infants with low delta APPG required ductal closure. Between the ages of 24 and 58 hours, five of the latter 26 infants had severe pulmonary hemorrhage, which was fatal in two cases. Severe (grade III or IV) intraventricular hemorrhage was seen in three infants with RDS and a low delta APPG; one of these infants also had severe pulmonary bleeding. Pneumothorax occurred in six infants with a low delta APPG. Infants without an increase in the APPG value during the first day of life required a significantly higher fraction of inspired oxygen. CONCLUSIONS: Noninvasive measurement of the change in APPG during the first day of life may provide a useful method for identifying infants with RDS at high risk of neonatal complications. PMID- 7472837 TI - Multiple organ involvement in perinatal asphyxia. AB - OBJECTIVES: (1) To evaluate the frequency and spectrum of severity of multisystem dysfunction after perinatal asphyxia and (2) to analyze the relationship between the clinical and biochemical markers of perinatal asphyxia and multiorgan involvement. STUDY DESIGN: Seventy-two consecutive term newborn infants with perinatal asphyxia were studied prospectively. Systematic neurologic, renal, pulmonary, cardiac, and gastrointestinal evaluations were performed. Involvement of each organ was classified as moderate or severe. RESULTS: Involvement of one or more organs occurred in 82% of the infants; the central nervous system (CNS) was most frequently involved (72%). Severe CNS injury (7 infants) always occurred with involvement of other organs, although moderate CNS involvement was isolated in 14 infants. Renal involvement occurred in 42%, pulmonary in 26%, cardiac in 29%, and gastrointestinal in 29% of the infants; 15% neonates had renal failure and 19% had respiratory failure. The Apgar scores at 1 and 5 minutes were the only perinatal factors related to the number of organs involved and the severity of involvement; the Apgar score at 5 minutes had the stronger independent association. No relationship or organ dysfunction was found with the umbilical cord arterial blood pH, meconium-stained amniotic fluid, umbilical cord abnormalities, presentation, or type of delivery. CONCLUSIONS: Our findings indicate that the Apgar score at 5 minutes, in infants who have other criteria for asphyxia, is the perinatal marker that may best identify infants at risk of organ dysfunction. PMID- 7472838 TI - Maternal and neonatal morbidity associated with in vitro fertilization. AB - OBJECTIVE: The purpose of this case-control study was to examine the maternal and neonatal morbidities associated with in vitro fertilization (IVF) in a single large teaching hospital. It was hypothesized that IVF mothers would have more perinatal complications and IVF infants would have higher mortality and morbidity rates than non-IVF control subjects. METHODS: One hundred forty-three gestations resulting from 101 IVF pregnancies, which included singletons (n = 62), twins (n = 72), and triplets (n = 9), were compared with equal numbers of non-IVF control subjects. Each pregnancy was matched by maternal age, race, insurance type, neonatal gender, order of gestation, order in delivery, and date of delivery (+/- 6 months). Among the 143 matched gestations, six IVF and seven control infants died, leaving 137 IVF and 136 control neonates for comparison. RESULTS: The IVF mothers had more pregnancy-induced hypertension (21% vs 4%), premature labor (44% vs 22%), labor induction (25% vs 1%), and preterm delivery (37% vs 21%). The IVF infant survivors had a lower mean (+/- SD) birth weight (2623 +/- 857 gm vs 3006 +/- 797 gm), more frequent occurrence of low birth weight (42% vs 27%), and shorter gestations (37 +/- 4 vs 38 +/- 3 weeks). The IVF infants had longer hospitalizations, more days of oxygen therapy, more days of continuous positive airway pressure, and increased prevalence of respiratory distress syndrome, patent ductus arteriosus, and sepsis. CONCLUSIONS: Couples who undergo IVF appear to be at increased risk of having low birth weight and preterm infants, and multiple gestations account for most of the neonatal morbidities. Both the mothers who conceive multiple gestations by means of IVF and their neonates are at an increased risk of having multiple morbidities. PMID- 7472839 TI - Diurnal variation in serum bilirubin concentrations in infants with neonatal jaundice. AB - We investigated whether there are independent intradaily changes in bilirubin levels in neonates. Healthy term newborn infants (n = 124) with neonatal jaundice bilirubin >/t171 mmol/L (10mg/dl) were studied for at least 3 consecutive days. Starting from the second day of life, consistent intradaily changes were observed in bilirubin levels (morning levels were greater than evening levels; p < 0.001), and body weight steadily increased. This diurnal variation in bilirubin levels should be considered in the follow-up and treatment of neonatal jaundice. PMID- 7472840 TI - Role of hemolysis in neonatal jaundice associated with glucose-6 phosphate dehydrogenase deficiency. AB - End-tidal carbon monoxide was measured in 108 newborn infants who had been screened for glucose-6-phosphate dehydrogenase (G6PD) deficiency. The mean +/- SD end-tidal carbon monoxide did not differ significantly between the G6PD-deficient and the normal neonates, 2.1 +/- 0.6 microliters/L and 2.0 +/- 0.5 microliters/L, respectively, within 12 hours of birth and 1.9 +/- 1.4 microliters/L and 1.5 +/- 0.7 microliters/L, respectively, at 48 to 72 hours after birth. On the basis of these measurements, hemolysis is not a sufficient explanation for jaundice in G6PD-deficient newborn infants in the transitional period. PMID- 7472841 TI - Favism in a female newborn infant whose mother ingested fava beans before delivery. AB - We describe a case of favism in a female newborn infant with glucose-6-phosphate dehydrogenase (G6PD) deficiency whose mother had ingested fava beans 5 days before delivery. At birth there were clinical and hematologic signs of hemolytic anemia, hemoglobinuria, and no blood group immunization. Study of the G6PD activity and 2-deoxy-glucose-6-phosphate utilization rate revealed that the infant and the mother were heterozygous for G6PD deficiency. PMID- 7472842 TI - Central hypothyroidism resulting from pituitary suppression and peripheral thyrotoxicosis in a premature infant born to a mother with Graves disease. AB - We observed the sequential changes in serum thyroid hormones and thyroid stimulating hormone receptor antibodies in an infant born at 30 weeks of gestation to a mother with florid Graves disease. Transient central hypothyroidism caused by pituitary suppression was observed after the resolution of peripheral thyrotoxicosis induced by thyroid-stimulating antibody. Central hypothyroidism became overt when the suppression of the pituitary gland after fetal thyrotoxicosis was combined with weak activity of thyroid-stimulating antibody after birth. PMID- 7472843 TI - Effects of L-thyroxine on serum lipid profiles in infants with congenital hypothyroidism. AB - Serum lipid profiles and the effects of thyroid hormone on them were studied in infants with congenital hypothyroidism. Serum total cholesterol, high-density lipoprotein cholesterol (HDLC), and low-density lipoprotein cholesterol levels were increased in infants with hypothyroidism;L-thyroxine therapy decreased the HDLC levels, leading to a reduction in serum concentrations of total cholesterol. It appears that thyroid hormone suppresses circulating HDLC, although the mechanism was not discovered in this study. PMID- 7472844 TI - Clearance of chronic hepatitis B virus infection in young children after alpha interferon treatment. AB - Nine children aged 18 months to 17 years (mean 5.7 years) with chronic hepatitis b virus infection and chronic active hepatitis were treated with 5 to 6 million units/m2 of body surface area of interferon -alpha 2b administered subcutaneously three times per week for 4 months (n = 1) or 6 months (n = 8). At 12 months after the start of therapy, six children less than 3 years of age responded to the treatment (three completely and three partially), whereas only one of three children older than 7 years of age responded. We conclude that IFN treatment may be effective in children with chronic HBV infection, especially when administered while they are young. PMID- 7472845 TI - Bone mineral density during treatment of central precocious puberty. AB - Treatment of adults with gonadotropin releasing hormone analogs has resulted in rapid loss in bone mineral density (BMD). We measured lumbar and femoral neck BMD by dual-energy x-ray absorptiometry during 2 years of depot leuprolide therapy in 13 girls (mean age, 7.5 years; mean bone age, 10.9 years). At baseline, BMD was elevated for age and concordant with the advanced skeletal age. During therapy with gonadotropin releasing hormone analog, BMD values increased and BMD standard deviation scores for age and skeletal age did not change. PMID- 7472846 TI - Prediction of outcome at school entry in neonatal intensive care unit survivors, with use of clinical and electrophysiologic techniques. AB - OBJECTIVE: To determine the predictive value of multimodality evoked potentials as well as the neonatal neurobehavioral assessment in neonatal intensive care unit survivors at school entry. STUDY DESIGN: In this prospective study, healthy (n = 24) and high-risk newborn infants (n = 78) were assessed in the newborn period with auditory brain-stem responses (ABRs) and somatosensory evoked potentials (SEPs), as well as the Einstein Neonatal Neurobehavioral Assessment Scale (ENNAS). Healthy and high-risk newborn infants were assessed in a blind fashion at 1, 3, and 5 years of age by a psychologist and a pediatric neurologist. Of those healthy (all 24) and high-risk newborn infants (72/78) with a neonatal ABR, SEP, or both, 62.5% were assessed at 5 years of age with the Wechsler Preschool and Primary Scale of intelligence, the Beery-Buktenica Test of Visual-Motor Integration, the Griffiths Locomotor Subscale, and neurologic examination. Chi-square analyses were carried out on neonatal and outcome measures, and sensitivity, specificity, and predictive values for each of the neonatal tests were ascertained. RESULTS: The ENNAS and the ABR had good negative predictive value for cognitive (85.7% to 93.3%), locomotor (83.3%), and visual motor (91.4% to 100%) performance. The ABR had good specificity, whereas the ENNAS was more sensitive. The SEP was an excellent prognostic tool, with high sensitivity (100%) and specificity (80% to 81.3%) for motor as well as cognitive domains. All infants with normal SEPs had favorable outcomes, whereas those with absent potentials did poorly. The SEP abnormalities were associated with findings on neurologic examination in all cases. CONCLUSIONS: Normal neonatal evoked potentials and ENNAS are associated with favorable outcomes. The SEP most accurately predicts neurodevelopmental status at school entry. PMID- 7472847 TI - Value of parents' estimates of children's developmental ages. AB - OBJECTIVE: To determine whether parents' estimates of children's developmental ages can function as a prescreening technique. DESIGN AND PARTICIPANTS: Parents of 234 children from birth to 77 months of age seeking well-child care in pediatric offices were queried in two separate studies. In the first study, parents were asked to give an estimate of their child's overall developmental age and, in the second study, to estimate ages in each of six developmental domains. Children were administered a range of screening measures of intelligence, speech language, and adoptive behavior. RESULTS: The overall age-estimate, if less than chronologic age, was 75% sensitive to likely developmental problems and, if equal to or greater than chronologic age, was 90% specific in identifying children likely to have typical development. Age estimates for each developmental domain were 81% sensitive to likely developmental problems if less than chronologic age in the domains of fine motor, language, grass motor, or behavior, and 62% specific if equal to or greater than chronologic age. Estimates at or below chronologic age in receptive language or personal-social domains were 90% sensitive and 43% specific in identifying likely behavior problems. There were no differences in the accuracy of parents estimates on the basis of children's age, gender, race, parents' level of education, or parenting experience. CONCLUSION: Parents' overall age-estimates provided a sensitive and specific indicator of global developmental status, but insufficient information about strengths and weaknesses to enable focused referrals for services. In contrast, discrete patterns of age estimates in each developmental domain sensitively discriminated children with developmental versus behavioral problems, although specificity was limited. Age estimates appear to be a potentially helpful method for identifying a subset of children in need of thorough screening, although further research is needed on a larger sample given diagnostic rather than screening tests. PMID- 7472848 TI - Paul Gyorgy, 1893-1976. PMID- 7472849 TI - Use of ipratropium for severe asthma. PMID- 7472850 TI - Circulatory effects of dopamine in neonates. PMID- 7472851 TI - Can treatment with recombinant granulocyte colony-stimulating factor transform neutropenia into leukemia? PMID- 7472852 TI - The evolution of virulence: a unifying link between parasitology and ecology. PMID- 7472853 TI - Lack of behavioral fever in nematode-parasitized Drosophila. AB - The parasitic nematode Howardula aoronymphium causes severe fertility reduction and sterility in females of several species of mycophagous Drosophila. Infected females of Drosophila putrida recover their fertility when kept at a postemergence temperature of 29 C but not when kept at 27 C or below. When given a choice of temperatures in a thermal gradient, neither this species, Drosophila falleni, nor Drosophila neotestacea manifested adaptive behavioral fever in response to parasitism. However, our conclusions concerning D. neotestacea and D. falleni remain tentative because of small sample sizes. PMID- 7472854 TI - Correlation between the level of infection with Bunodera inconstans and Neoechinorhynchus rutili and behavioral intensity in female brook sticklebacks. AB - We investigated the influence of 2 intestinal helminths, Bunodera inconstans and Neoechinorhynchus rutili, on the development of female behavioral and morphological characters in brook sticklebacks. The number of N. rutili per female is positively correlated with standard length. Both the number of B. inconstans per female and the number of N. rutili per female are negatively correlated with the intensity of a female's aggressive response to an intruding female and are positively correlated with the intensity and duration of female courtship. The results of this study indicate that infection with N. rutili and B. inconstans may influence a female brook stickleback's ability either to compete for a mate or to sample from a large subset of territorial males. PMID- 7472855 TI - Cyclic AMP mediates change in superoxide dismutase activity to monitor host parasite interaction in Leishmania donovani. AB - The present study is designed to understand the role of cyclic AMP (cAMP) in host parasite interaction involving Leishmania donovani, the causative agent for Kala azar. When Leishmania promastigotes or macrophages were pretreated with dibutyryl cAMP or theophylline and epinephrine, which are well defined initiators for cAMP release, a key enzyme of the oxygen defense system, superoxide dismutase (SOD), was inhibited. At the same time, parasite interaction was considerably reduced to the level of 54.5% and 46.2%, respectively, for pretreated promastigotes. Internalization of the organisms in phagolysosomes was similarly affected. Dibutyryl cAMP-treated promastigotes in the presence of SOD, on the other hand, restored in vitro infection to the normal level. At least 50% less cAMP entered into Leishmania promastigotes when SOD was added to the incubation system containing dibutyryl cAMP. Data reveal that cAMP perturbs the Leishmania macrophage interaction through inhibition of SOD, pointing to the importance of a promastigote enzyme for the survival of this pathogen within phagolysosomes. PMID- 7472856 TI - Infectivity of Trypanosoma rangeli in a promonocytic mammalian cell line. AB - The existence of an intracellular stage of Trypanosoma rangeli in the vertebrate host was evaluated by experimental infection of the U937 histiocytic cell line with the San Agustin strain and the Ub66-5b clone. The identity of the parasites at the beginning and end of the experiments was confirmed through biological behavior in the vector and mammal hosts, isoenzymes, polymerase chain reaction (PCR), and monoclonal antibodies. Infectivity to U937 cells of T. rangeli obtained from culture and salivary glands was evaluated under different experimental conditions. These included 34 C vs. 37 C, opsonized vs. nonopsonized parasites, and 2, 4, 6, 24, 48, and 72 hr of cell-parasite contact. Trypanosoma rangeli adopted a characteristic nondividing amastigote-like form within U937 cells, which was different in size (P = 0.001) from Trypanosoma cruzi amastigotes. Culture forms of T. rangeli were more infective than parasites from salivary glands (P = 0.049) but were less infective than T. cruzi (P = 0.0001). Variations in temperature (34-37 C) and complement opsonization did not affect infectivity. Viability of intracellular forms was confirmed by feeding Rhodnius prolixus with T. rangeli-infected cells. Resistance of T. rangeli to the intracellular milieu could be an important mechanism in producing chronic infections in mammals and in the infection of triatomines. PMID- 7472857 TI - Behaviors associated with egg and parasite deposition by gravid and Lambornella clarki-infected Aedes sierrensis. AB - Egg- and Lambornella clarki-deposition behaviors by the treehole mosquito Aedes sierrensis were monitored inside laboratory deposition containers using a video camera. Gravid and infected mosquitoes were seen probing the substrate and in the water with their abdominal tips. Both eggs and parasites were recovered from the containers. Gravid and infected mosquitoes performed the same or similar types of behaviors, but gravid mosquitoes spent significantly higher percentages of time engaged in exploratory probing movements than did infected mosquitoes. Gravid mosquitoes also spent significantly less percentage time resting and more percentage time grooming than did infected mosquitoes. It appears that, compared to gravid mosquitoes, infected mosquitoes spent less relative time in certain behaviors that may not benefit the parasite. PMID- 7472858 TI - Some effects of sarcoptic mange on dogs. AB - Sequential changes in pathology were examined for scabies-infested dogs to determine the effects of infestation with Sarcoptes scabiei var. canis. During 8 wk of infestation with S. scabiei, the progression of the disease was evaluated weekly by skin scrape, clinical examination, and blood analyses. At 8 wk, selected organs were microscopically examined for histopathology. All infested dogs developed an advanced level of scabies infestation by 8 wk. Of the 36 blood parameters evaluated, only values for erythrocyte sedimentation rate (ESR) deviated significantly from the normal ranges for dogs. However, infested dogs had significantly (P < 0.01) lower average hemoglobin and hematocrit concentrations after 8 wk of infestation compared to their values prior to infestation or to the values for the control dogs. Red blood cell levels for infested dogs dropped significantly (P < 0.01) from preinfestation concentrations by week 8. Conversely, by 8 wk total white blood cell and neutrophil concentrations were significantly (P < 0.01) greater than uninfested controls. Also, whereas average eosinophil concentrations were not statistically different for infested dogs compared to controls, some individual infested dogs exhibited eosinophilia at 4-8 wk of infestation. The ESRs for infested dogs were significantly (P < 0.01) greater at week 6 and 8 than for experimental dogs prior to infestation or control dogs. All parameters except neutrophils had returned to preinfestation levels by 2 wk after treatment for scabies. Neutrophil concentrations were no longer significantly different by 4 wk posttreatment. There were no significant differences in serum enzyme, biochemical and electrolyte concentrations between infested and control dogs.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7472859 TI - Avian malaria seroprevalence in Jackass penguins (Spheniscus demersus) in South Africa. AB - Blood samples of 191 adult Jackass penguins (Spheniscus demersus) from South Africa were tested by indirect enzyme-linked immunosorbent assays (ELISA) with Plasmodium falciparum antigen (R32tet32) for avian malaria antibodies (Ab). The samples originated from free-ranging penguins from offshore islands and southern coast colonies (3 groups, n = 110), from 2 penguin groups (n = 66) rescued after offshore oil-spill contamination and rehabilitated at the Rescue Station of the South African National Foundation for the Conservation of Coastal Birds (SANCCOB) in Cape Town, and from SANCCOB-resident penguins (n = 15). The total average malaria Ab seroprevalence was 39%, and the mean malaria seropositivity ranged from 20% to 62% among the 6 S. demersus groups, with a mean of 55% for oiled penguins, and 31% for the remaining birds. The total mean absorbance value was 0.57 for ELISA-positive penguins, 0.43 for birds kept at SANCCOB facilities, and 0.70 for the penguins from wild colonies. The 2 groups of oiled penguins exhibited higher malaria Ab seroprevalence (38% and 62%) than the 3 groups of non oiled birds (29%, 33% and 35%). Malaria Ab seroprevalence of free-ranging penguins was significantly lower (P < 0.01) than in the 3 groups of birds at SANCCOB facilities and did not differ significantly among 3 wild penguin colonies. PMID- 7472860 TI - Schistosoma japonicum, zoophilic strain, in Oncomelania hupensis chiui and O. H. formosana: miracidial penetration and comparative histology. AB - Miracidial penetration and intramolluscan development of a zoophilic strain of Schistosoma japonicum were studied in the highly susceptible Oncomelania hupensis chiui and a highly resistant population of Oncomelania hupensis formosana. When exposed to 20 miracidia, an average of 3.6 miracidia penetrated O. h. chiui and 3.4 penetrated O. h. formosana. The distribution of sporocysts in the 2 snail subspecies was similar: 70.4% migrated to the cephalopedal sinus, 14.2% to the anterior viscera, and 3.1-4.9% to the posterior viscera, tentacle, mantle collar, and foot. In O. h. chiui only 1 out of 118 sporocysts degenerated and there was no host cellular response. In O. h. formosana, all the sporocysts survived for 12 hr, but by 5 days postinfection (DPI) all were dead and by 10 DPI they had disintegrated completely; the encapsulation was encountered in only 3 out of 78 sporocysts or 3 out of 23 snails, suggesting that it played a minor role in the defense against S. japonicum. All of the O. h. formosana snails of 10 and 15 DPI showed pronounced leukocytosis and elevated phagocytosis in the heart and increased cell debris in tissue spaces, which coincided with the disappearance of the parasites. It is suggested that these phenomena were provoked in response to degradation products released by the dead parasites. PMID- 7472861 TI - Production of Schistosoma mansoni daughter sporocysts from mother sporocysts maintained in synxenic culture with Biomphalaria glabrata embryonic (Bge) cells. AB - In vitro production of Schistosoma mansoni daughter sporocysts (DS) from miracidium-derived mother sporocysts (MS) was achieved by synxenic larval cultivation with cells of the Biomphalaria glabrata embryonic (Bge) cell line. The in vitro growth and viability of MS cocultured with Bge cells or in Bge cell conditioned medium were significantly extended beyond that of larvae cultured in fresh medium alone. However, complete DS development and emergence from MS were achieved only in the presence of Bge cells. Introduction of either miracidia or previously transformed MS onto Bge cell monolayers resulted in an initial attachment of cultured cells to sporocysts, followed by a gradual encapsulation of larvae by multiple layers of Bge cells. Sporocysts and their encapsulating cells eventually formed large cellular aggregates, within which MS increased 4 fold in size during the first 20 days of cultivation. The timing of in vitro DS development was somewhat variable; however, in general, early embryo formation, i.e., germ cell aggregates with surrounding primitive epithelium, was first detected at 15-20 days of culture, whereas motile, intra-MS daughter stages were seen at 25-30 days and thereafter. Mature, first generation DS, measuring 136 +/- 46 microns long by 22 +/- 6 microns wide, emerged from MS starting at approximately 30-45 days of initial cultivation. Although the basic morphology and size of emergent, in vitro-derived DS were comparable to those propagated in vivo, there was a large reduction in the in vitro reproductive capacity of the MS and a delay in DS culture development.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7472862 TI - Sources and reservoirs of Toxoplasma gondii infection on 47 swine farms in Illinois. AB - Field studies were conducted on 47 swine farms in Illinois during 1992 and 1993 to identify sources and reservoirs of Toxoplasma gondii infection. Blood samples were obtained from swine and from trapped wildlife. Serum antibodies to T. gondii were determined using the modified agglutination test, incorporating mercaptoethanol. Antibodies to T. gondii (titer > or = 25) were found in 97 of 4,252 (2.3%) finishing pigs, 395 of 2,617 (15.1%) sows, 267 of 391 (68.3%) cats, 126 of 188 (67.0%) raccoons, 7 of 18 (38.9%) skunks, 29 of 128 opossums (22.7%), 6 of 95 (6.3%) rats, 3 of 61 (4.9%) white-footed mice (Peromyscus sp.), and 26 of 1,243 (2.1%) house mice (Mus musculus). Brains and hearts of rodents trapped on the farm were bioassayed in mice for the presence of T. gondii. Toxoplasma gondii was recovered from tissues of 7 of 1,502 (0.5%) house mice, 2 of 67 (3.0%) white footed mice, and 1 of 107 (0.9%) rats. Feces of 274 cats trapped on the farm and samples of feed, water, and soil were bioassayed in mice for the presence of T. gondii oocysts. Toxoplasma gondii was isolated from 2 of 491 (0.4%) feed samples, 1 of 79 (1.3%) soil samples, and 5 of 274 (1.8%) samples of cat feces. All mammalian species examined were reservoirs of T. gondii infection. All farms had evidence of T. gondii infection either by detection of antibodies in swine or other mammalian species, or by detection of oocysts, or by recovery from rodents by bioassay. The possibility of transmission of T. gondii to swine via consumption of rodents, feed, and soil was confirmed. PMID- 7472863 TI - Vertical transmission of Neospora caninum in mice. AB - Herein we report a murine model to examine transplacental transmission, transmammary transmission, or both, of Neospora caninum. Prevalence of transplacental transmission in outbred Swiss-Webster mouse pups was 85%, with 11 of 13 litters containing at least 1 transplacentally infected pup. Sixty-two percent of litters born to experimentally infected dams contained 85% or more transplacentally infected pups. Numbers of pups congenitally infected per litter was higher if dams were inoculated during the first half of pregnancy. Transplacental transmission decreased to 25% when a singly infected dam delivered a second litter. Transmammary transmission was observed in 1 of 51 pups suckling dams experimentally infected 5, 10, or 15 days postparturition. No pups were infected when cross-fostered onto chronically infected dams. PMID- 7472864 TI - Egg deposition of the intestinal nematode Heterakis spumosa: stimulation by soluble colon factors of mice. AB - The nematode Heterakis spumosa lives and develops in the upper colon of mice. The colon mucosa contains soluble factors that significantly increase release of eggs by female H. spumosa in vitro. Egg release is stimulated more when colon factors are prepared from mice harboring adult H. spumosa. Also, soluble factors stimulating egg release can be prepared from homogenates of H. spumosa but are not contained in H. spumosa-conditioned medium. Moreover, soluble factors from the small intestine of mice infected with Heligmosomoides polygyrus also stimulate egg release by H. spumosa females in vitro to about the same extent as soluble factors from the colon of H. spumosa-infected mice. Our data suggest that colon factors are involved in the regulation of egg deposition of H. spumosa, the efficiency of these factors is increased by intestinal inflammation, and that these factors are accumulated by H. spumosa worms. PMID- 7472865 TI - Risk factors for transmission of Toxoplasma gondii on swine farms in Illinois. AB - Two epidemiologic studies of risk factors for transmission of Toxoplasma gondii to swine were conducted for farms in Illinois. The first study was a cross sectional survey of swine farms from the state of Illinois pseudorabies testing program, in which farm owners or managers were interviewed by telephone regarding presence of risk factors for transmission of T. gondii on the farm. There were 123 farms surveyed that provided blood samples for at least 30 sows. The mean sow seroprevalence was 19.5% (median = 10.0%). Multiple regression analysis of the association of sow seroprevalence with outdoor housing of sows, cat access to sow areas, number of sows, open feed storage and water delivery, delayed removal of carcasses, and presence of rodents on the farm indicated that higher sow seroprevalence was associated with cat access to sows (P = 0.009) and fewer sows in the herd (P = 0.05). The second study was a field investigation of 47 swine farms (37 from the cross-sectional study). Data collection included obtaining blood samples from swine, cats, and rodents, and fecal samples from cats, heart and brain tissue from rodents, and feed, water, and soil samples for T. gondii examination. The risk of T. gondii transmission from cats and rodents to sows and finishing pigs was evaluated, taking into account housing conditions and herd size. Multiple regression analysis indicated that T. gondii seroprevalence in finishing pigs increased with more seropositive juvenile cats on the farm (P < 0.0001) and higher seroprevalence in house mice (P = 0.0023).(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7472866 TI - Serum-free culturing of adult Schistosoma mansoni in dialysis bags for the production of excretory/secretory antigens. AB - Antigenic excretory/secretory (E/S) products from Schistosoma mansoni are potentially important in the development of diagnostic assays used to detect circulating antigens in schistosomiasis. The E/S products to be used as antigen(s) for this development must, by necessity, be free of exogenous proteins. The ability to extend serum-free in vitro culture of adult worms is, therefore, essential. Adult worms were perfused from mice, washed in serum-free RPMI-1640 with antibiotics, and placed in sterile dialysis bags, molecular weight cut-off 10 kDa, at a concentration of 100 worms in 1 ml of serum-free, supplemented RPMI-1640. Each bag was then placed in a flask of supplemented RPMI 1640 with 10% fetal calf serum in a humidified incubator at 37 C, 7% CO2. At days 1, 3, 5, 8, and 12, worms were collected; E/S culture medium was analyzed by sodium dodecyl sulfate polyacrylamide gel electrophoresis. Worm survival rates were 85% after 1 day in culture, dropping gradually to 65% on day 8, and then to 38% on day 12. Silver stain for total protein and immunoblot exposed to positive human infection serum showed E/S culture media from days 3 and 5 having the least complex banding pattern. The quantitative specific activity of E/S, as measured by antigen-limiting Falcon assay screening test system-enzyme-linked immunosorbent assays against human infection serum, indicates E/S antigenicity closely follows the attrition of worms and, therefore, may be directed against the release of somatic antigens by dead worms. Culturing S. mansoni in dialysis tubing is useful in deriving E/S products. PMID- 7472867 TI - Control of ciliary activities of Schistosoma mansoni miracidia using Triton extracted parasites. AB - Extraction with 0.04% (w/v) Triton X-100 for 3 min removed the cell membrane from the locomotory cilia of Schistosoma mansoni miracidia while leaving the motile apparatus apparently intact. Immediately after Triton-extracted miracidia were treated by the reactivating solution containing ATP and magnesium ions (Mg2+) at pH 8.1, nearly 100% of Triton-extracted miracidia showed the ciliary beating and swam forward in a manner resembling that of a normal miracidium. In the standard reactivating solution (2 mM ATP, 2 mM Mg2+, pH 8.1), Triton-extracted miracidia swam at a speed of 580 microns/sec; the comparable value for live miracidia in dechlorinated tap water was 2,200 microns/sec. The swimming velocity of Triton extracted miracidia was dependent on ATP and Mg2+ concentration, pH, and salinity. In a solution containing 0.9% NaCl, Triton-extracted miracidia were not reactivated. Among the nucleotides tested, only ATP was found to induce a significant amount of ciliary motility. In terms of divalent cation specificity, only Mg2+ was capable of producing normal motility. Barium and calcium ions (at 0.5 mM CaCl2) also were capable of activating ciliary motility but were less effective stimulants than Mg2+. However, in 1 mM CaCl2, no ciliary reactivation was observed and cilia became detached from the body surface of the miracidia. Vanadium inhibited ATP-reactivated ciliary beating of Triton-extracted miracidia. PMID- 7472868 TI - Redescription of Argulus melanostictus (Branchiura: Argulidae), a parasite of California grunion (Leuresthes tenuis: Atherinidae), with notes regarding chemical control of A. melanostictus in a captive host population. AB - Adult male and female Argulus melanostictus Wilson, 1935 are redescribed based on detailed examinations of a syntype and recently obtained specimens of both sexes collected from California grunion, Leuresthes tenuis, captured in nearshore Pacific waters at Monterey, California. A 14-16-hr seawater bath containing 0.5 microliter/L trichlorfon administered once weekly for 3 wk killed A. melanostictus while not noticeably harming grunion. PMID- 7472869 TI - Serendipidae Evenhuis, 1994 (Insecta: Diptera) and Serendipidae Brooks and Barriga, 1995 (Platyhelminthes: Eucestoda): proposed removal of homonymy. PMID- 7472870 TI - Philometra (Ranjhinema) salgadoi n. sp. (Nematoda: Philometridae) from the ocular cavity of the red grouper Epinephelus morio (Pisces: Serranidae) from the yucatan peninsula, Mexico. AB - A new nematode species, Philometra (Ranjhinema) salgadoi n. sp., is described from females found in the ocular cavity of the red grouper (Epinephelus morio) from the Mexican coast of the Gulf of Mexico (type locality Telchac, Yucatan). The nematode is characterized by the presence of 8 conspicuously large, crescent shaped, fleshy cephalic papillae of the external circle and 2 small, subterminal papilla-like projections on the caudal end. Another philometrid species, Philometroides belousae n. sp., is established by indication for P. parasiluri (Yamaguti, 1935) sensu Ermolenko, 1984; its adult females are parasitic in the subcutaneous tissues of Percottus glehni in the Russian Far East. PMID- 7472871 TI - The genus Centrorhynchus (Acanthocephala) in North America with description of Centrorhynchus robustus n. sp., redescription of Centrorhynchus conspectus, and a key to species. AB - Reexamination of the type series of Centrorhynchus wardae Holloway, 1958, and Centrorhynchus conspectus Van Cleave and Pratt, 1940, revealed no consistent difference between the 2 purported species; therefore, C. wardae is considered a junior synonym of C. conspectus. Specimens from additional collections are consistent with this view and provided adequate material for redescription of the species. Centrorhynchus robustus n. sp. resembles C. conspectus in the number of hooks per Salient differences between the 2 species are size and body shape, with C. robustus having a robust trunk with the widest diameter anteriad and C. conspectus being elongate with no region of the trunk consistently widest. Additionally, a conspicuous posteriad swelling exhibited by females of C. conspectus is lacking in C. robustus. Centrorhynchus kuntzi is reported for the first time from Montana and Nebraska, and C. conspectus is reported for the first time from an eastern screech owl, Otus asio. PMID- 7472872 TI - Proteocephalus chamelensis n. sp. (Cestoda: Proteocephalidae) from the "guavina" Gobiomorus maculatus (Osteichthyes: Eleotrididae) in Chamela Bay, Jalisco, Mexico. AB - A new species of Proteocephalus inhabiting the eleotridid estuarine fish Gobiomorus maculatus is described from Chamela Bay, Jalisco State, Mexico. The new species and P. pugetensis are nearly identical in strobila length, presence of a vestigial apical organ, having vaginae only anterior to the cirrus sac, an average of 4-7 uterine diverticula per proglottis, and a ratio of cirrus sac length to proglottis width averaging approximately 1:4. Proteocephalus pugetensis differs from the new species by having an average of 35 rather than 68 testes per proglottis, vitelline follicles terminating posteriorly at the level of the anterior margin of the ovary rather than at the level of the posterior margin, and equatorial genital pores rather than genital pores located in the anterior 1/4 of the proglottis. Finally, Proteocephalus pugetensis occurs from estuarine habitats along the Pacific coast of the United States; the new species was collected from an estuarine habitat on the Pacific coast of Mexico. PMID- 7472873 TI - Potentiation of ionophorous anticoccidials with duokvin: battery trials against Cryptosporidium baileyi in chickens. AB - Three battery tests, employing 444 Arbor Acres chickens, were conducted to study the anticryptosporidial efficacy of the polyether ionophorous anticoccidials alone and in combination with a new dihydroquinoline antioxidant, duokvin. The following efficacy percentages for the ionophores at the recommended level were established on the basis of oocyst output: lasalocid 45, monensin 37, semduramicin 29, narasin 23, and maduramicin and salinomycin 21. The efficacy of the ionophores, with the exception of lasalocid, could be increased to 36-88% by combination with duokvin. However, these combinations resulted in a significant weight gain reduction due to toxic interaction. Efficacy showed a strong negative correlation with body weight gain and bursa weight, suggesting an indirect mode of action against the parasite. It has been concluded that neither ionophorous anticoccidials nor their combinations with duokvin can be recommended for chemoprophylaxis against avian cryptosporidiosis. PMID- 7472874 TI - Detection of Trichinella spiralis muscle larvae in naturally infected horses. AB - Human trichinellosis outbreaks related to horsemeat consumption have been reported in France and Italy in recent years. In order to determine if Trichinella is present in horses slaughtered at an abattoir in the State of Mexico, diaphragm muscle tissue samples (22-37 g) from 80 horses were examined by artificial digestion. Four of these samples had larvae that were characterized as Trichinella sp. by morphological criteria and as Trichinella spiralis by the polymerase chain reaction. PMID- 7472875 TI - Experimental oral inoculations in birds to evaluate potential definitive hosts of Neospora caninum. AB - Experimental oral inoculations to evaluate potential definitive hosts of Neospora caninum were conducted by feeding infected rodent tissues to 9 carnivorous birds of 4 species. Birds included 2 red-tailed hawks (Buteo jamaicensis), 2 turkey vultures (Cathartes aura), 2 barn owls (Tyto alba), and 3 American crows (Corvus brachyrhynchus). The rodents (mice or rats) had been inoculated with 100,000 culture-derived tachyzoites of N. caninum 1-6 mo before feeding to the birds. Fecal samples were collected from each bird daily for 1 mo after feeding rodents and examined for oocysts by fecal flotation. In addition, processed aliquots from all avian fecal samples were fed to BALB/c mice. Five weeks after feeding, mice were bled and sera were tested for antibodies against N. caninum. One to two months later, mice were killed and brain tissue was examined microscopically for protozoal cysts. While occasional oocysts were found in avian fecal samples, these were likely not N. caninum because they were not infective to BALB/c mice. It was concluded that the bird species tested are not likely to be definitive hosts of N. caninum. PMID- 7472876 TI - Recovery of helminth eggs from archeological excavations of the Grand Louvre (Paris, France). AB - Archeological excavations at the Grand Louvre in Paris, France, during 1985-1990, provided an opportunity for paleoparasitological investigation at various ancient sites of habitation, dating from the 11th to the 16th centuries. Eggs of helminths were recovered by means of sedimentation and flotation. The genera of helminths represented were Ascaris, Trichuris, Heterakis, Metastrongylus, Fasciola, and Dicrocoelium. PMID- 7472877 TI - Bot fly (Cuterebra fontinella fontinella) parasitism of cotton mice (Peromyscus gossypinus) on St. Catherines Island, Georgia. AB - From February 1993 through October 1994, 382 cotton mice (Peromyscus gossypinus) were live-trapped and examined for bot fly (Cuterebra fontinella fontinella) larvae. Thirty-nine (10%) of the mice were parasitized by a total of 44 bot fly larvae. All bots were situated in the inguinal or lower abdominal region of the mice. Monthly data strongly suggested that this bot is bivoltine at this latitude with distinct summer and winter population peaks. Prevalence of parasitism was highest in June 1994 when 56% of 16 mice examined were infested with a mean intensity of 1.3 bots. Adult flies were successfully reared from 10 bots that exited from mice; the time period (58-98 days, mean = 79.5) that these specimens spent in the soil as pupae further supports the contention that C. f. fontinella is bivoltine on St. Catherines Island. PMID- 7472878 TI - Survival and infectivity of Babesia in blood maintained at 25 C and 2-4 C. AB - Babesia microti-infected blood was stored at room temperature (approximately 25 C) or refrigerated (4 C) for 30 days. To assess viability of the parasites after storage at these 2 temperatures, a 0.25-ml aliquot was inoculated into each of 2 hamsters in 2 separate experiments at days 3, 7, 10, 14, 17, 21, 25, and 30. Blood films were prepared and examined weekly for the presence of parasites from all hamsters. Of hamsters inoculated with blood held at room temperature, only those inoculated at day 3 became positive, whereas 4/4 hamsters inoculated with refrigerated blood on day 17 became parasitemic and 1/4 hamsters inoculated with blood held for 21 days became parasitemic. These results indicate that under blood banking conditions, this intracellular protozoan parasite can remain infective and transfusion-acquired infection with this parasite could occur throughout most of the time that blood is normally stored. PMID- 7472879 TI - The role of tadpoles and frogs as paratenic hosts in the life cycle of Dracunculus insignis (Nematoda: Dracunculoidea). AB - The possibility exists that paratenic hosts play a role in the life cycle of various Dracunculus species. In the present study, we established that tadpoles of 2 genera of frogs (Xenopus and Rana) were capable of ingesting copepods infected with third-stage larvae (L3) of Dracunculus insignis. Once ingested, the L3s migrated from the gut to the somatic tissues of the tadpoles. In Xenopus, the dracunculid larvae persisted through the metamorphosis of the tadpoles into adult frogs. These observations confirm the concept that paratenic hosts, such as tadpoles or frogs, may be important means of transporting infective larvae of Dracunculus species up the food chain and facilitate transmission to the definitive hosts. PMID- 7472880 TI - Activity of benzimidazoles against cryptosporidiosis in neonatal BALB/c mice. AB - The need for an effective compound for the prevention and treatment of cryptosporidiosis in humans and animals has led to the testing of benzimidazoles based on reports that albendazole was clinically effective against related protozoan parasites causing microsporidiosis in humans. Albendazole and other benzimidazole derivatives were tested for prophylactic efficacy against cryptosporidiosis at dosage levels 1-3x the levels found effective for treatment of cattle or sheep for helminth infections. Daily dosage levels of thiabendazole, parbendazole, oxibendazole, mebendazole, and albendazole, as high as 200, 30, 10, 15, and 15 mg/kg of body weight, respectively, were not efficacious in neonatal mice. Although the number of parasites in histologic sections of intestine from mice mediated with 15 mg albendazole/kg of body weight was significantly lower than in unmedicated control mice, suggesting activity against the parasite, a high percentage of epithelial cells in the medicated mice were infected. PMID- 7472881 TI - New distribution record of Taenia mustelae Gmelin, 1790 (Cestoda) from the least weasel Mustela nivalis in Hokkaido, Japan. PMID- 7472882 TI - Density-dependent effects on Steinernema glaseri (Nematoda: Steinernematidae) within an insect host. AB - Increasing densities of Steinernema glaseri infective juveniles (IJs) in soil affected penetration efficiency and reproduction of the nematodes in larvae of the greater wax moth Galleria mellonella. The penetration efficiency and the proportion of penetrated IJs developing to adults decreased significantly with increasing numbers of IJs present in the soil and entering the hosts, respectively. The number of progeny produced/host cadaver initially increased, with the highest production being between 20.7 +/- 3.3 and 58.0 +/- 6.6 IJs established/host +/- SE and decreased at higher densities. Above 184.4 +/- 37.7 IJs established/host, no progeny emerged from the cadavers. PMID- 7472883 TI - Gnathostoma spinigerum: growth and development of third-stage larvae in vitro. AB - Advanced third-stage larvae of Gnathostoma spinigerum were cultured in RPMI-1640, with various supplements at 37 C under 5% CO2 in air for 300 days. The most suitable medium supplement for worm development was 10% fetal calf serum, 1% dog serum, and 0.25% dog hemolysate. After approximately 180 days of cultivation, some larvae molted to the fourth stage as distinguished by 8 transverse rows of cephalic hooklets and well differentiated sex organs. The maximum body length and width of these larvae were 18.6 mm and 1.1 mm, respectively. Six of 50 larvae (12%) developed to the fourth stage, with a 32% survival rate at the end of cultivation. Although the highest survival rate (70%) of the worms was observed in the medium supplemented with 25 mM NaHCO3, only 4% developed into fourth stage larvae. The addition of fetal calf serum, dog serum, and dog hemolysate was essential for growth and development. PMID- 7472886 TI - The first record of fish as paratenic hosts of Falcaustra spp. (Nematoda: Kathlaniidae). AB - Two types of third-stage nematode larvae (most of which were encapsulated) were collected from the abdominal cavity and liver of the freshwater fishes Etheostoma fonticola, Etheostoma lepidotum, Cichlasoma cyanoguttatum, Lepomis auritus, Lepomis sp., and Gambusia affinis in Texas and are referred to the genus Falcaustra. This is the first record of Falcaustra larvae from fishes, and their presence may reflect the importance of fishes as paratenic hosts of turtle parasites in this genus. PMID- 7472884 TI - Coccidia (Apicomplexa: Eimeriidae) from the big bend slider, Trachemys gaigeae (Testudines: Emydidae), in New Mexico. AB - Twenty-nine Big Bend sliders Trachemys gaigeae (Hartweg, 1934) were collected from Socorro County, New Mexico, and their feces examined for coccidial parasites. Three (10%) of the turtles were found to be infected with at least 1 coccidian. Seven Eimeria spp. (E. chrysemydis, E. graptemydos, E. marginata, E. pseudemydis, E. pseudogeographica, E. stylosa, and E. trachemydis) were harbored by T. gaigeae. All represent new host and distributional records for these previously described coccidians. In addition, a single sympatric western painted turtle (Chrysemys picta bellii) harbored E. chrysemydis, E. graptemydos, and E. trachemydis. The latter coccidian is reported for the first time from C. picta bellii. PMID- 7472885 TI - Sensitivity of antigen ELISA test for detecting Trypanosoma evansi antigen in horses in the subtropical area of Argentina. AB - The sensitivity of an antigen detection enzyme immunoassay (Ag-ELISA) based on a Trypanosoma brucei group-specific monoclonal antibody was evaluated to detect circulating Trypanosoma evansi antigen in horse sera. Three horses and 2 mules were experimentally infected with T. evansi. Circulating antigens were detected on 7 and 21 days postinfection. Antigen levels increased during the course of the illness and remained high even when parasitemia was low or when parasites could not be detected. Antigens were cleared from serum when drug treatment was effective but persisted when it was not. In 6 outbreaks of "mal de caderas" involving 125 horses, T. evansi was found in 78 horses using standard parasite detection methods and antigenemia was detected in 58 of them (74%). The Ag-ELISA sensitivity rate varied between 63% and 100% for the 6 different outbreaks. A combination of Ag-ELISA and parasitologic methods diagnosed a total of 93 infected animals. These results show that the Ag-ELISA test is useful both to diagnose T. evansi and to assess the efficacy of drug treatment in horses. PMID- 7472887 TI - A coprological survey of intestinal parasites of wild lions (Panthera leo) in the Serengeti and the Ngorongoro Crater, Tanzania, east Africa. AB - Fecal examination on 112 and a subset of 58 wild lions (Panthera leo) in the Serengeti National Park and the Ngorongoro Crater, Tanzania, East Africa revealed eggs, larvae, and protozoan cysts of 15 parasite taxa. The most prevalent were Spirometra sp. (63% prevalence), Taeniidae (58%), Ancylostoma sp. (56%), and a coccidian (53%). Three parasites were considered spurious. Of the hosts, 97.3% were infected with at least 1 species of parasite. Individual parasite taxa were aggregated among hosts. PMID- 7472888 TI - Invasion of Tilapia mossambica (cichlidae) viscera by the monogenean Enterogyrus cichlidarum. AB - The monopisthocotylean monogenean Enterogyrus cichlidarum, which normally inhabits the stomach and anterior intestinal lumina of tilapine cichlids, invaded the internal organs of juvenile Mozambique tilapia (Tilapia mossambica). Adult worms infected the intestinal mucosa, peritoneal cavity, liver, heart, blood vessels, swimbladder, and braincase. Most of the parasites were adults and contained eggs. Both adults and eggs incited a mononuclear inflammatory response. Infections were associated with chronic morbidity and mortality. Although affected fishes were overcrowded, the precise stress responsible for such an unusually invasive event is unknown. PMID- 7472889 TI - Host-induced morphological variability in adult Posthodiplostomum minimum (Digenea: Neodiplostomidae). AB - Posthodiplostomum minimum adults were collected from natural hosts at Lake Patzcuaro, Michoacan, Mexico and from experimentally infected hosts. Intraspecific variation was measured using principal component analysis (PCA). Morphological traits such as general body shape, size, and the specific location of certain distinctive organs and structures showed great variation among hosts as described by PCA. These results extend previous studies on host-induced variation in P. minimum in showing that some conservative traits such as vitelline follicle distribution in fore-and hindbody, could be used for an extensive and complete revision of the genus Posthodiplostomum. PMID- 7472890 TI - Hammondia hammondi cysts in cell cultures. AB - In an attempt to obtain the continuous development of Hammondia hammondi in vitro, culture of the parasite was performed in 3 cell lines. Although organisms were present in culture fluids of feline kidney cells (CRFK) for as long as 3 mo, continuous culture was not possible. However, for the first time, cysts of H. hammondi were observed in cell culture from day 6 after inoculation of sporozoites. Ultrastructure of the H. hammondi cysts was similar to that observed for in vitro-obtained Toxoplasma gondii cysts. Feeding a cat with these in vitro developed cysts resulted in oocyst shedding 5 days after ingestion. PMID- 7472891 TI - Repeated measurements of blood parasite levels reveal limited ability for host recovery in the common lizard (Lacerta vivipara). AB - I investigated longitudinal measures of haemogregarine load in the host lizard Lacerta vivipara over 2 yr (1992-1993). Lizards with heavy parasite infections in 1992 were still suffering from heavy parasite infections in 1993. Such data show that lizards have limited ability to recover from high levels of haemogregarine infection. Modifications of host behavior and of life-history traits may represent a more effective and less costly response to haemogregarine infection in this species. PMID- 7472892 TI - Age-dependent resistance to Cryptosporidium baileyi infection in chickens. AB - The age-dependent aspects of resistance to Cryptosporidium baileyi were studied in chickens orally inoculated with C. baileyi oocysts at 1 and 9 wk of age. The prepatent period was significantly shorter and the patency was significantly longer in younger birds. Chickens infected at 1 wk of age excreted 3 times the number of oocysts excreted by those inoculated at 9 wk of age. There was a good correlation between the length of the patent period and the total oocyst output of chickens. These results indicate that both innate and acquired resistance to C. baileyi are age-related. The numbers of animals needed per experimental group to estimate the length of the prepatent period, the patency, and the total oocyst output were assessed. Relatively small numbers of animals are required for the estimation of the mean length of the prepatent period, in contrast with the large numbers of birds needed for the estimation of the mean length of the patent period or that of the total oocyst excretion. PMID- 7472893 TI - Killing of Schistosoma mansoni sporocysts in Biomphalaria glabrata implanted with amoebocyte-producing organ allografts from resistant snails. AB - Schistosome-susceptible National Institutes of Health (NIH) albino Biomphalaria glabrata were implanted with the amoebocyte-producing organ (APO) from 4 types of donors: (1) exposed-resistant (eR), i.e., schistosome-resistant 13-16-R1 snails that had been exposed to miracidia of Schistosoma mansoni 30 or more days previously in order to verify their resistance, (2) exposed-susceptible (eS), i.e., NIH albino snails that had been similarly exposed to miracidia, (3) unexposed-resistant (uR), and (4) unexposed-susceptible (uS). Allograft recipients, along with unimplanted NIH albino and 13-16-R1 controls (cS and cR, respectively), were then challenged with 100 miracidia each of Schistosoma mansoni at 14-15 days postimplantation. Histological sections of tentacles fixed at 3 days postchallenge (PC) showed significantly fewer normal sporocysts and more numerous developmentally retarded sporocysts in cR snails than in the other 5 treatment groups, and significantly more killed sporocysts in both cR snails and recipients of eR APOs than in the other 4 groups. In addition, the histological condition of eR allografts in both unchallenged (at 1, 3, 7, 10, and 14 days postimplantation) and schistosome-challenged (at 3 days PC) NIH albino recipients was examined. Viable hematopoietic cells were found in 96% of implants, and in 86% of implants low numbers of mitotic figures were found among these cells, although no increased mitotic activity occurred in challenged recipients. These data suggest that lowered susceptibility to infection with S. mansoni in recipients of APO allografts results primarily from hemocyte-mediated resistance. PMID- 7472894 TI - Activity of the molluscicidal plant Millettia thonningii (Leguminosae) toward Biomphalaria glabrata eggs. AB - A dichloromethane extract of the seeds of the molluscicidal west African legume Millettia thonningii was tested for ovicidal activity toward Biomphalaria glabrata egg masses. The extract was found to be highly ovicidal at concentrations as low as 5 mg/L. Embryonic development of the snails in egg masses was monitored using photomicrographs from which embryonic diameters were estimated. Such measurements revealed that ovicidal effects were developmentally stage specific and normally induced a curtailment of development during the gastrula to trochophore transition. PMID- 7472895 TI - Parasitology year 2000. AB - We predict that in order for parasitology to thrive by the year 2000 the various subdisciplines of evolution, ecology, biosystematics, and genetics must develop holistic approaches and use parasite models to answer basic biological questions. The students of tomorrow must work as part of a multidisciplinary team; and their questions and answers must be conceptually integrated into the broader biological framework of evolution and ecology. PMID- 7472896 TI - Microbes to fight microbes: a not so novel approach to controlling diarrheal disease. PMID- 7472897 TI - New views on the pathogenesis of kwashiorkor: methionine and other amino acids. PMID- 7472898 TI - Etiology and epidemiology of persistent diarrhea in northeastern Brazil: a hospital-based, prospective, case-control study. AB - With the improved therapy for acute diarrhea, persistent diarrhea (> 14 days) is emerging as a major problem in developing countries. However, the etiologies and pathogenesis of persistent diarrhea remain poorly understood. We conducted a prospective case-control study in children < 3 years old presenting to the hospital with persistent diarrhea in Fortaleza, Brazil. Over the study period (August 1988 to March 1991), 56 children seen with persistent diarrhea, 52 children seen with acute diarrhea, and 42 controls attending the same hospital/clinic for illnesses other than diarrhea were enrolled. A potential pathogen was found in 91% of children with persistent diarrhea and 90% of those with acute diarrhea versus 45% of controls (both p's < 0.01). Thirty-four percent of persistent (19/56) and 38% of acute (20/52) diarrhea cases versus 2% (1/42) of controls (both p's < 0.01) had multiple pathogens. Enteroaggregative Escherichia coli (EAggEC) were found in 68% (38/56) of children with persistent diarrhea versus 31% (13/42) of controls (p < 0.01) and in 46% (24/52) of those with acute diarrhea. Furthermore, when the EAggEC were subdivided into aggregative adherence (AA) gene probe positive (18/56; 32%) and negative (20/56; 36%), both subgroups were still significantly different from controls [6/42 (14%) and 7/42 (17%), respectively; both p's < 0.05].(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7472899 TI - Effectiveness of nasogastric rehydration in hospitalized children with acute diarrhea. AB - The American Academy of Pediatrics recommends oral rehydration and early refeeding for management of infants with diarrhea and mild to moderate dehydration. However, intravenous rehydration is still widely used for treatment of infants hospitalized for dehydration. The administration of oral rehydration solution via continuous infusion through a nasogastric tube facilitates its delivery in hospitalized children. The purpose of this study is to compare intravenous and nasogastric rehydration in children hospitalized for mild to moderate dehydration. Infants who failed attempts at oral rehydration and were hospitalized for dehydration due to acute diarrheal illness were randomized to receive intravenous or nasogastric rehydration. Following rehydration, infants received soy formula and a maintenance oral electrolyte solution to replace ongoing stool losses, as directed by the attending physician. Patients were discharged from the hospital once oral feeding was tolerated, and the vomiting and diarrhea resolved. Twenty-four patients, from 2 to 19 months of age, were enrolled in the study. Rehydration was successful in 11 of 12 patients in the nasogastric rehydration group and in all 12 patients who received intravenous rehydration. The degree of dehydration, severity of vomiting and diarrhea, and duration of rehydration were similar in both groups. The duration and cost of hospitalization were less for patients receiving nasogastric rehydration compared to those who were rehydrated intravenously. Rehydration by infusion of oral rehydration solution via a nasogastric tube is a safe and effective treatment for infants with mild to moderate dehydration. Rehydration with infusion of oral rehydration solution through a nasogastric tube should be considered for in patient management of infants with diarrhea. PMID- 7472900 TI - Highly destructive perianal disease in children with Crohn's disease. AB - The perianal complications of Crohn's disease (CD) seen in children and adolescents include skin tags, anal fissures, fistulae, and abscesses. While these lesions are often chronic and variably responsive to medical therapy, only rarely are they severely destructive. In this report, we characterize the frequency, severity, and clinical course of a highly destructive form of perianal disease (HDPD) that we have noted in a number of children and adolescents with Crohn's disease. A database containing records from 350 children with inflammatory bowel disease was reviewed to identify all children with CD treated between 1970 and 1993. For each, the occurrence or absence of significant perianal pathology, including fistula, abscess, and HDPD, was determined. Pertinent clinical details were recorded for all patients. In addition, the clinical characteristics of those children with HDPD were compiled, and the courses of those with HDPD characterized. A search of the database identified 230 children and adolescents with CD followed for a total of 1,518 patient years. Sixty-seven of these patients (29% of the CD population) had significant perianal pathology. This included 6 with HDPD, 8 with complicated fistulae [rectourethroperineal (1), rectovaginal (1), rectolabial (2), and multiple communicating perineal (4)], and 53 with simple perianal fistulae or abscesses. All six with HDPD had deeply destructive perineal ulcerations, marked undermining of the perineal and perirectal tissues, and copious exudate, and often there was a deeply cleaved or fileted perineum on separating the buttocks. Two children with HDPD had fecal incontinence. PMID- 7472901 TI - Oxygen radical scavengers are protective against indomethacin-induced intestinal ulceration in the rat. AB - Nonsteroidal antiinflammatory drugs (NSAIDs) are widely used and may cause small intestinal inflammation and damage. Reactive oxygen metabolites are involved in various gastrointestinal inflammatory processes, but there is little information about their role in small intestinal mucosal damage induced by NSAIDs. We studied the effect of the oxygen radical scavengers superoxide dismutase (SOD), catalase (CAT), and allopurinol (ALLO) on indomethacin (INDO)-induced intestinal ulceration in the rat. Ulceration was produced by s.c. injection of 30 mg/kg of INDO 30 min after refeeding 24 h-fasted rats. Total ulcer area was measured 24 h after INDO administration. Study groups each consisted of eight animals which received either i.p. CAT, SOD, or both together, at a dosage of 5,000 U/kg each. All drugs were divided into five doses, given once an hour over a 4-h period, starting at the time of INDO injection. Another group received 100 mg/kg ALLO in two doses. Total ulcer area was reduced by SOD from 228 +/- 12 (sq mm, mean +/- SEM) to 153 +/- 12 (p < 0.001), by CAT to 179 +/- 13 (p < 0.01), and by both together to 95 +/- 5 (p < 0.0001). ALLO administration reduced the total ulcer area to 176 +/- 7 (p < 0.003). The protective effect of oxyradical scavengers supports the hypothesis that oxygen radicals are involved in the pathogenesis of INDO-induced small intestinal ulceration in the rat. PMID- 7472902 TI - Early proliferative events following intestinal resection in the rat. AB - Partial resection of the small intestine results in compensatory proliferation and adaptation in the remaining small intestinal mucosa. The molecular mechanisms governing the proliferative response are not known, nor has the timing of events associated with proliferation been adequately defined, particularly during the period just after resection. We designed experiments to characterize early (within 24 h) proliferative events associated with proximal intestinal resection and sought to determine the cell type that first responds to proliferative stimuli. Twenty-one day old male Sprague-Dawley rats underwent a 70% proximal intestinal resection or transection (control). Poly(A) RNA was isolated from the distal (ileal) remnants. Northern blots showed a marked induction of the immediate early genes zif-268, nup-475, and c-myc 1-3 h following resection, but not following transection. Immunohistochemical analysis of c-myc expression in ileal crypt epithelial cells showed a biphasic induction that was most marked 6 h after resection and less prominent 24 h after resection. Immunostaining with 5 bromodeoxyuridine (5-BrdU) was restricted to ileal crypt nuclei and was maximal 24 h after resection. All these events were observed in the absence of nutrient intake. Taken together, these data show that a potent nutrient-independent stimulus for intestinal epithelial cell proliferation occurs within minutes of partial small intestinal resection and that the first targets of this stimulus are crypt epithelial cells in the residual intestine. PMID- 7472903 TI - Effect of early dietary deficiency in polyunsaturated fatty acids on two lectin binding sites in the small intestine of postweanling rats. AB - This study was designed to determine whether dietary lipids influence the development of intestinal cell glycosylation, in relationship to diet-induced changes in phospholipid fatty acid composition. The ability of two different lectins, wheat germ agglutinin (WGA) and Maackia amurensis agglutinin (MAA), to combine specifically with particular carbohydrate residues was used to investigate the surface characteristics of epithelial cells of rats fed different dietary lipids from birth to 6 weeks of age. Diets contained 5% (weight) peanut oil (PO), rich in n-6 fatty acids; salmon oil (SO), rich in n-3 fatty acids; hydrogenated palm oil (HPO), deficient in both n-6 and n-3 fatty acids or a PO and rapeseed oil (RO) mixture (PRO), the control diet. Pieces of jejunal and ileal villi were excised from postweanling rats and prepared for lectin histochemical study. Concurrently, epithelial cells were removed from jejunal and ileal segments for determining their phospholipid fatty acid compositions. Polyunsaturated fatty acid (PUFA) deficiency was evidenced in the HPO group by the appearance of eicosatrienoic acid (20:3n-9) in both jejunal and ileal phospholipids, which paralleled the decrease in arachidonic acid content. Accretion of 18:1n-9 and 20:3n-9 in cell phospholipids of group HPO was not sufficient to match the unsaturation level in rats fed nonhydrogenated vegetable oils (PRO, PO) or fish oil (SO). The lectin histochemical study showed that WGA strongly labelled the brush border membrane microvilli whereas binding of MAA was specific to goblet cells and mucus. Regardless of the type of diet, WGA binding was weaker in the ileum than in the jejunum. In comparison to all other groups, WGA-labelling of villi was less intense in the jejunum and disappeared almost completely in the ileum of HPO-fed rats. Although SO- and PO-fed rats had, respectively, very low and high ratios of n-6 to n-3 in their intestinal phospholipids, binding of WGA in both groups was not markedly different from that in the control (PRO). MAA-labelling was very intense in jejunal and ileal villi of n-3-fed (SO) rats, whereas it was strongly attenuated in the n-3- and n-6 deficient (HPO) group. These results suggest that intestinal glycosyltransferase activities involved in cell differentiation were altered relative to the overall unsaturation index of dietary fatty acids. Alterations of epithelial glycosylation mainly resulted from a drop in total n-6 and n-3 fatty acids, although it may be speculated that there is a specific effect of n-3 fatty acids. PMID- 7472904 TI - Effect of a fermented infant formula containing viable bifidobacteria on the fecal flora composition and pH of healthy full-term infants. AB - We assessed the growth, tolerance, and acceptability as well as fecal flora composition and stool pH of 20 healthy full-term infants fed with a fermented whey-adapted infant formula containing viable bifidobacteria (10(6)/g of powder) during the first 2 months of life. This fermented infant formula, first biologically acidified by Streptococcus thermophilus and Lactobacillus helveticus, was compared to a whey-adapted, nonacidified, low-phosphate infant formula in a double-blind, randomized controlled study. The results were compared to a control group (n = 14) of fully breast-fed infants. The fermented whey adapted formula containing viable bifidobacteria induced a prevalence of colonization with bifidobacteria at 1 month of age similar to that of breast-fed infants (12/20 versus 8/14) but significantly higher than in the group fed the standard infant formula (4/20). The mean bacterial count of bifidobacteria was similar in all colonized infants; however, fecal pH was significantly lower in the breast-fed infants than in the nonacidified bottle-fed infants. This kind of infant formula was well tolerated and promoted a normal growth during the first 2 months. PMID- 7472906 TI - Determination of iron absorption using erythrocyte iron incorporation of two stable isotopes of iron (57Fe and 58Fe) in very low birthweight premature infants. AB - Due to limited erythropoiesis, iron needs for the premature infant during the first 2 months of life are low. With the potential for increased use of erythropoietin in the preterm infant, iron requirements may become markedly higher. The ability of the preterm infant to absorb iron, therefore, becomes crucial. Previous studies using single stable isotopes of iron without metabolic balances were unable to quantitatively determine iron absorption since the percentage of absorbed iron appearing in the erythrocyte could not be measured. The objective of the current study was to determine iron absorption using the method of erythrocyte iron incorporation of two stable isotopes of iron given by the enteral and parenteral route to very low birth weight (VLBW) infants. Results obtained were compared to iron absorption values from studies using single isotopes and metabolic balance techniques. Six VLBW premature infants (gestational age 26.8 +/- 1.7 weeks, postnatal age 3.6 +/- 1.9 weeks, birth weight 863 +/- 117 g) were studied. Iron dosages were (i.v.) 0.15 mg/kg (57FeSO4) and (enteral) 1.5 mg/kg (58FeSO4). Erythrocyte isotope ratios, 57Fe/54Fe and 58Fe/54Fe, were determined by inductively coupled plasma mass spectrometry (ICP/MS) from single blood samples (100 microliters) collected before and after concurrent enteral (58Fe) and parenteral (57Fe) administration of isotopes. Only 17.8% of the i.v.-infused iron dose was incorporated into hemoglobin on day 15. Using a correction factor based on the percentage of i.v. iron (57Fe) incorporated into erythrocytes, the corrected incorporation of 58Fe was calculated to be 26.3 +/- 13.0% of the enteral dose of 58FeSO4. PMID- 7472905 TI - Effect of three levels of vitamin D intake in preterm infants receiving high mineral-containing milk. AB - Very low-birth weight (VLBW) infants fed high-calcium and high-phosphorus (10.74 and 6.93 mmol/MJ; 180 and 90 mg/100 kcal, respectively) infant formulas were randomized to one of three levels of vitamin D intake to approximate 200, 400, and 800 IU/day. Sixty-two infants completed the study (24 to 29 days), with actual mean daily vitamin D intakes of 161, 361, and 766 IU, respectively. Outcomes were not different by group: gains in body weight, length and head circumference, serum calcium, magnesium, phosphorus, alkaline phosphatase, osteocalcin, 25-hydroxyvitamin D, 1,25-dihydroxyvitamin D, and urine calcium:creatinine and magnesium:creatinine ratios. There were no radiographic fractures and/or rickets. A subset of 19 infants was followed between 173 and 380 days to determine descriptively if there was any delayed effect of earlier manipulation of vitamin D intake. They were fed standard infant formulas with a vitamin D content of 400 to 420 IU/L. No significant differences were present among the three groups, and data were combined. Serum 25-hydroxyvitamin D increased (p < 0.05), osteocalcin decreased (p < 0.05), and 1,25-dihydroxyvitamin D decreased (p = 0.06) at follow-up. Thus, for VLBZW infants fed high-calcium and high-phosphorus milk, an average daily vitamin D intake as low as 160 IU maintains normal and stable vitamin D status and normal physical growth, biochemical and hormonal indexes of bone mineral metabolism, and skeletal radiographs versus randomized infants receiving about 400 or 800 IU of vitamin D per day. On follow-up, vitamin D status remained normal for > or = 6 months while infants received < 400 IU of vitamin D per day. PMID- 7472907 TI - Effects of a low birthweight infant formula containing human milk levels of docosahexaenoic and arachidonic acids. AB - Long-chain (LC) polyunsaturated fatty acids (PUFA) (LCP) are considered conditionally essential nutrients for low birth weight infants (LBWI). Therefore, enrichment of LBWI formulae with metabolites both linoleic (omega-6) and alpha linolenic (omega-3) acids at levels typical for human milk has been recommended. However, previous feeding trials with LCP-enriched formulae evaluated only a dietary supplementation with omega-3 LCP from fish oils alone or with both omega 3 and omega-6 LCP at levels considerably lower than usual human milk contents. We studied the effects of an LBWI formula providing the major omega-3 and omega-6 LCP, docosahexaenoic and arachidonic acids, in amounts similar to those in average human milk. Twenty-seven LBWIs were enrolled in this study when they tolerated full enteral feeding (> or = 130 ml milk/kg/day). Infants either received their own mother's milk (n = 8, birthweight 1218 +/- 146 g, gestational age 30.2 +/- 1.5 weeks, mean +/- SD) fortified with protein and minerals (FM-85, Nestle Ag, Munchen, Germany; dosage 5 g/100 ml milk) or were randomly assigned to blinded batches of an LBWI formula (Prematil, Milupa AG, Friedrichsdorf, Germany) without LCP (n = 10, 1280 +/- 229 g, 31.1 +/- 3.1 weeks) or with LCP (n = 9, 1253 +/- 334 g, 30.4 +/- 3.3 wks.). During the study period of 21 days, the three feeding groups did not differ in growth and feeding tolerances as assessed by occurrence of gastric residuals, spitting, or abdominal distention; however, firms stools were noted more frequently in the two formula groups.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7472908 TI - Tracer kinetic studies on a methionine-supplemented soy-based infant formula using 1-13C- and 15N-methionine as tracers. AB - A tracer-kinetic study using 1-13C- and 15N-labeled L-methionine was conducted in order to measure the retention rate of free methionine added to commercially produced soy-based infant formulas. Twelve male infants, fed on a soy formula, received a single-pulse labeling by oral administration of L-1-13C-methionine (5 mg/kg) and L-15N-methionine (10 mg/kg). The abundance of expired 13C-labeled CO2 was measured up to 7 h after administration at 15-, 30-, and 60-min intervals. Additionally, enrichment of total 15N and 15N in urinary ammonia were determined up to 48 h after administration. Retention rates of the labeled carboxyl group amounted to an average of 91.2% (SD 4.1) of the intake. A similar retention rate was measured for the 15N-label of methionine (90.0%, SD 4.3). The data point at the efficacy of methionine supplementation of soy-based infant formulas. PMID- 7472909 TI - Utility of duplex Doppler ultrasound in evaluating portal hypertension in children. AB - We measured the maximal velocity of the blood flow in the main portal trunk by duplex Doppler ultrasound in children suffering from a variety of liver diseases. The maximal velocity of the main portal vein in children with chronic active hepatitis and liver cirrhosis was decreased significantly as compared to those in control children (p < 0.01). The maximal velocity of the main portal trunk and K indocyanine green (ICG) had no significant correlation (r = 0.25, n = 27). The patient with the lowest portal velocity had esophageal and gastric varices with red color sign. We conclude that the measurement of the maximal velocity of the main portal trunk with duplex Doppler ultrasound is useful in evaluating portal hypertension in children. PMID- 7472910 TI - Plasma endothelin-1 levels of children with cirrhosis. AB - We measured plasma concentration of endothelin-1 in three children with Byler's disease, five with biliary atresia after portoenterostomy, and nine controls. No patients had ascites or hepatorenal syndrome. Plasma endothelin-1 levels were significantly higher in patients with Byler's disease than in the controls (5.19 +/- 0.90 versus 1.81 +/- 0.19 pg/ml, respectively; p < 0.01), but were normal in operated biliary atresia. Urinary concentrations of N-acetyl-beta-D glucosaminidase (NAG) were significantly higher in the patients with Byler's disease than in controls. Plasma endothelin-1 level correlated significantly with serum concentration of bile acid (r = 0.91; p < 0.01) and urinary concentration of NAG (r = 0.92; p < 0.01). We conclude that plasma endothelin-1 levels are high in patients with severe biliary cirrhosis and that endothelin-1 may partially contribute to development of renal injury in cirrhosis. PMID- 7472912 TI - Multiple chronic hemorrhagic ulcers of the small intestine in children: a report of two cases. PMID- 7472911 TI - Treatment of recurrent Clostridium difficile colitis with Lactobacillus GG. PMID- 7472913 TI - Ileal and colonic ulceration due to enterobiasis. PMID- 7472914 TI - Cow's milk protein sensitivity: a possible cause of nephrotic syndrome in early infancy. PMID- 7472915 TI - Serological diagnosis of celiac disease. PMID- 7472916 TI - Response to "The clinical significance of a prolonged stable pH around 4.0 in 24 H pH monitoring". PMID- 7472917 TI - Hepatitis C virus infection in pediatric cholestatic diseases. PMID- 7472918 TI - Heating characteristics of the TRIPAS hyperthermia system for deep seated malignancy. AB - A deep heating hyperthermia device TRIPAS (a triapplicator system) consisting of three independent, dielectrically loaded horn applicators operating in phase at 300 MHz was investigated. The heating characteristics produced by this hyperthermia system were analyzed by means of thermochromic liquid-crystal cards and a modified CDRH (Center for Devices and Regulatory Health) elliptical phantom. Both homogenous and inhomogeneous phantoms were used, simulating high and low permittivity tissues (muscle and lung). These equivalent tissues were made of polyacrylamide gel. The semiquantitative heating pattern analysis showed a central heating of 1/3 of maximum heating at a depth of 10 cm in both homogenous (muscle) and heterogenous (muscle/fat) phantoms. Also more uniform temperature/SAR distributions were generated in muscle equivalent material than those in lung. PMID- 7472919 TI - Alleged health effects of electromagnetic fields: the misconceptions continue. AB - Exposures to electric or magnetic fields (including microwaves), in residential or occupational settings, have been reported to be associated with health problems, particularly cancer and reproductive mishaps. Misconceptions about these alleged effects continue to be published in the medical and scientific literature, as well as in popular press reports. This paper is the third in a series of papers challenging invalid statements relating to these alleged effects. Reports dealing with epidemiologic studies (including exposure assessment, consistency, and public policy implications), the use of biomarkers, and risk assessment are analyzed. PMID- 7472920 TI - Adaptation of rabbit small intestinal brush-border membrane enzymes after extensive bowel resection. AB - Short lengths of small intestine may be resected without significantly affecting the digestive and absorptive capacity; however, extensive resection produces varying degrees of malnutrition. This study was undertaken to define the adaptive changes in the remaining small intestine of two of the jejunal and ileal mucosal brush-border membrane enzymes after extensive small bowel resection in rabbits. Thirty adult New Zealand White rabbits underwent a 50% to 60% jejuno-ileal enterectomy with end-to-end anastomosis. Maltase activity (UE/g protein) increased from 152 (preoperative) to 392 at 3 weeks in the proximal segment and from 85 to 259 in the distal segment; these levels decreased to 222 and 155 in the respective segments at 6 weeks. AOP activity (UE/g protein) increased from 154 (preoperative) to 171 in the proximal segment and 171 to 256 in the distal segment at 3 weeks, and was 131 and 288 in the respective segments at 6 weeks. This marked increase in the mucosal brush-border enzymatic activities at 3 weeks postoperatively was associated with a 28% increase in bowel length. By 6 weeks the enzyme levels had decreased slightly; however, there was a persistent 41% increase in bowel length over that immediately postoperation. The mucosal surface area increased from 295 mm2 immediately postoperation to 5,337 mm2 at 3 weeks and 7,635 mm2 at 6 weeks, a 250% increase. The authors conclude that there is an immediate compensatory increase in villus length as well as brush-border enzymatic expression in the remaining intestine that gradually declines as the small intestinal surface area continues to increase and the bowel lengthens with time. PMID- 7472921 TI - Impaired survival and growth in immunosuppressed young rats with lethal short gut syndrome and a small bowel transplant: an effect of cyclosporine. AB - Neonates and growing individuals have increased nutritional demands as compared with adults. To determine the functional ability of an intestinal graft to allow survival and growth, an otherwise lethal short gut model should be used (resection of both the entire small bowel and the cecum). In this study the authors investigated the survival and growth in young rats (80 to 125 g) with this lethal short gut syndrome (SGS) and either syngeneic or allogeneic segmental small bowel transplantation (SBTx). Additionally they sought to determine the effect of therapeutical doses of cyclosporine (CyA) in young, growing rats. To avoid total parenteral nutrition in rats undergoing SBTx, surgery was carried out in two steps: after segmental SBTx of a 25-cm jejunal graft, SGS was created 2 weeks later. Lewis rats underwent 1: Syngeneic segmental SBTx + SGS (n = 7); 2: Allogeneic segmental SBTx (donor: Lewis Brown Norway F1) + SGS + CyA (15 mg/kg/d for 7 days, then every other day for 21 days) (n = 9); 3: Syngeneic segmental SBTx + SGS + CyA as in group 2 (n = 5); 4: SGS alone (n = 5): 5: small bowel resection alone (n = 5); 6: sham laparotomy twice (n = 5); 7: sham laparotomy twice + CyA as in group 2 (n = 6). Weight, general condition, and nutritional serum variables were followed up regularly for 4 months. Rats with resection of small bowel survived but did not grow. Rats with small bowel resection + cecectomy died within 5 days.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7472922 TI - Association of imperforate anus with occult spinal dysraphism. AB - Neurological deficits may occur in some children with imperforate anus produced by unrecognized spinal cord anomalies amenable to neurosurgical correction. Although this association has been well described, its frequency has not been defined. The authors prospectively studied 45 infants with imperforate anus from 1985 to 1992 to establish the frequency of this association. Eighteen infants (40%) had high imperforate anus, 24 had low imperforate anus (53%), and 3 had anal stenosis (7%). All underwent radiographic spinal cord imaging in early infancy, which consisted of radiographs (45), ultrasound (26), magnetic resonance imaging (29), computed tomography (1), and myelography (1). Thirty-three percent of the patients with imperforate anus had bony sacral abnormalities. Eight infants were discovered to have spinal cord abnormalities. Two of those infants had obvious myelomeningocele at birth, and 6 infants (14%) had occult spinal dysraphism consisting of tethered cord with or without associated lipoma of the cord. There was no correlation between the presence of tethered cord and the gender of the infants, the type of malformation (high versus low imperforate anus), or the presence of sacral dysplasia (Fisher's exact test). Five of the infants who were found to have tethered cord underwent neurosurgical intervention and repair of the defect, and 1 infant was followed up clinically. The mean follow-up time of all the infants was 2.5 +/- 0.9 (+/- SD) years. No neurological sequelae occurred in the infants who had surgical correction. The frequency of occult spinal dysraphism with imperforate anus was 14%. It was independent of the type of imperforate anus or the coexistence of bony sacral anomalies.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7472923 TI - Cyclic GMP relaxes the internal anal sphincter in Hirschsprung's disease. AB - In Hirschsprung's disease (HD), the aganglionic colon and internal anal sphincter (IAS) fail to relax. Aganglionic colon of HD patients relaxes in response to exogenous nitric oxide (NO), whereas the IAS from HD patients does not relax. The authors hypothesized that the failure of IAS relaxation is caused by a local deficiency of cyclic guanosine monophosphate (cGMP), the final metabolite in NO mediated smooth muscle relaxation. To test this hypothesis, the authors measured the isometric tension of smooth muscle strips taken from the IAS and aganglionic colon of patients with HD before and after exposure to cGMP and compared this with ganglionic colon and IAS from normal controls. In HD patients both the IAS and aganglionic colon relaxed in response to cGMP (P < .05). The amount of relaxation observed in both the aganglionic colon and IAS was comparable to that measured in the normal controls. The observation that exogenous cGMP relaxes the IAS, whereas exogenous NO does not, suggests that mechanisms for relaxation may be different than those in the aganglionic colon and may explain persistent IAS dysfunction after resection of aganglionic colon. The defect of the IAS in HD may be the inability of the NO/cGMP pathway to induce smooth muscle cell relaxation rather than a defect in the smooth muscle cell. PMID- 7472924 TI - Primary laparoscopic pull-through for Hirschsprung's disease in infants and children. AB - Between November 1993 and September 1994, 12 primary laparoscopic colon pull through procedures were performed in infants and children. The patients' ages ranged from 3 days to 6 years. The primary diagnosis in all 12 patients was Hirschsprung's disease. All children had their operations without construction of preoperative or postoperative colostomy. Three 5-mm abdominal wall ports were used for access to the peritoneal cavity. The sigmoid colon and proximal rectum were mobilized laparoscopically. A submucosal sleeve was developed transanally to meet the dissection from above. The colon was then pulled down in continuity, divided above the transition zone, and secured to the anal mucosa 5 to 10 mm above the pectinate line. Mean postoperative stay was 4 days. Laparoscopic visualization provides clear delineation of pelvic structures even in small infants. Laparoscopic pull-through requires no more time than similar open procedures, averaging just over 2 hours. The morbidities associated with colostomy formation and closure and the inconvenience of colostomy care are avoided with a one-stage technique. These benefits combined with the advantages of minimally invasive surgery make primary laparoscopic pull-through a potential advance in the surgical treatment of Hirschsprung's disease. PMID- 7472925 TI - Enterocolitis associated with Hirschsprung's disease: a clinical histopathological correlative study. AB - Enterocolitis associated with Hirschsprung's disease (HEC) remains a major source of morbidity and even mortality, both before and after definitive surgical treatment. This study was undertaken to investigate whether histopathologic mucosal changes, in the absence of clinical manifestations of HEC, could predict the subsequent development of this complication. The clinical data and histopathology of 25 patients who eventually developed clinical HEC were compared with a control group of 25 age- and sex-matched patients with Hirschsprung's disease (HD) but with no clinical HEC either preoperatively or postoperatively. The histopathologic findings of tissue obtained by rectal biopsy or during laparotomy were graded from 0 to V according to severity and compared with the eventual clinical course. This study showed that (1) histopathologic findings of HEC appear to predict the clinical development, but not the severity, of future episodes of clinical HEC; (2) a histological grade of > or = II (> or = 2 crypt abscesses per high power field) should raise suspicion for subsequent occurrence of HEC, whereas a grade of > or = III (multiple crypt abscesses) places a child at high risk for development of clinical HEC; and (3) contrary to what is generally thought, histopathologic changes of HEC occur in both ganglionic and aganglionic segments. Based on these findings, it is recommended that histopathologic documentation of HEC and its grade should be an integral part of the tissue diagnosis of HD. PMID- 7472926 TI - Creation of myelomeningocele in utero: a model of functional damage from spinal cord exposure in fetal sheep. AB - A recent study in human fetuses with myelomeningocele (MMC) suggested that the primary malformation is not neural but a failed closure of the posterior vertebral column and paraspinal soft tissue, which leads to exposure and secondary destruction of the spinal cord. The goal of this study was to test whether chronic exposure of the normal spinal cord to the amniotic space produces a lesion similar to human MMC. In fetal sheep at 75 days' gestation (group A) and 60 days' gestation (group B) (term = 150 days), the lumbar spinal cord was exposed to the amniotic cavity by excising skin and paraspinal soft tissues, and by performing a laminectomy. Some animals from both groups were fetectomized and assessed morphologically at 100 days' gestation. The remainder were delivered near term and assessed clinically, electrophysiologically, and morphologically. In group A, all animals showed MMC-type pathology. The exposed spinal cord was herniated out of the spinal canal and rested on the dorsal membranes of a cystic sac. The neural tissue was stretched and flattened out. Histologically, the hallmarks of the spinal cord were not discernable and the cytoarchitecture was lost. These changes were less severe at 100 days than at term. The three survivors in group A were paraplegic. In group B, the two survivors and two fetuses harvested at 100 days had healed skin wounds and near normal spinal cord histology. The other animal harvested at 100 days had a MMC-type lesion with less severe histological changes. The two survivors had a mild paraparesis.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7472927 TI - Conservative surgical management of vaginal and vulvar pediatric rhabdomyosarcoma: a report from the Intergroup Rhabdomyosarcoma Study III. AB - The current management of pediatric vaginal and vulvar rhabdomyosarcoma (RMS) uses cytoreductive chemotherapy before surgical intervention. During the Intergroup Rhabdomyosarcoma Study (IRS)-III (1984 to 1988), 27 evaluable patients were entered on a preoperative chemotherapy protocol. Among 24 patients with vaginal primaries, 20 had an initial biopsy with gross residual (group III), 3 had resection with positive margins (group IIA) and 1 had metastatic disease (group IV). At subsequent surgery, 7 patients underwent partial or complete vaginectomy and 6 of them had no viable tumor identified in the specimen. Only 1 of these 7 patients underwent a cystectomy, whereas 5 underwent hysterectomy. Seventeen patients in this group have no evidence of disease 66 to 108 months after diagnosis; 2 died of chemotoxicity and 1 of unknown causes after achieving a complete response. Ten of these 17 patients were treated with biopsy and chemotherapy only. Four of these 10 had radiotherapy as well. All group IIA patients have no evidence of disease. The 1 group IV patient had rapidly progressive disease with early death. This primary chemotherapy protocol resulted in less need for operative intervention or irradiation compared with previous experience. There was no local recurrence, and 20 of 24 patients remain continuously relapse-free with no evidence of disease. All 3 patients with vulvar primaries were treated by wide local excision and chemotherapy and have no evidence of disease. Conservative surgical intervention for vaginal RMS with primary chemotherapy and adjunctive radiation when necessary appears to result in excellent disease-free survival. PMID- 7472928 TI - Neonatal soft tissue sarcomas: the influence of pathology on treatment and survival. Children's Cancer Group Surgical Committee. AB - INTRODUCTION: A multi-institutional study was conducted by the Children's Cancer Group (CCG) to evaluate all soft tissue sarcomas diagnosed within the first month of life. METHODS: A retrospective study by 11 CCG institutions of patient records from 1971 to 1991 were reviewed for demographic data, pathology, therapy, and outcome. RESULTS: 32 neonates with soft tissue sarcomas were identified. There were 21 boys and 11 girls. Pathology was equally divided into three groups: Congenital fibrosarcoma (CFS) (12), rhabdomyosarcoma (RMS) (11), and non-RMS soft tissue sarcomas (NRSTS) (9). Anatomic sites consisted of head/neck (11), extremity (9), trunk (8), pelvis (3), and unknown (2). Overall survival rate was 59% (19/32). CONCLUSION: Soft tissue sarcomas in the neonate comprise three general groups with survival rates dependent on pathology and extent of disease. PMID- 7472929 TI - Favorable outcome in children with Beckwith-Wiedemann syndrome and intraabdominal malignant tumors. AB - Children with Beckwith-Wiedemann syndrome (BWS) have an increased risk of developing Wilms' tumors, hepatoblastomas, and adrenal tumors. This study evaluates disease-free survival in children with BWS and intraabdominal tumors. Sixteen tumors occurred in 13 children with BWS (8 boys, 5 girls). Diagnoses included Wilms' tumor (10) (2 bilateral, 20%), hepatoblastoma (2), bladder rhabdomyosarcoma (1), and adrenal cortical tumor (1). In the 10 children with Wilms' tumor, the average age at diagnosis was 3.5 years (range, 7 months to 5 years). Nine of 10 had initial tumor resection, chemotherapy, and radiation therapy (when indicated). One child with bilateral disease had tumor biopsy, chemotherapy, and partial nephrectomy. Tumors were classified as stage I (5), stage II (2), stage IV (1) and stage V (2), all with favorable histology. Disease free survival rate was 100% with median follow-up of 9 years (range, 4 to 22 years). One patient had a left adrenal tumor detected during screening sonography 11 years after Wilms' tumor resection. Two infants with advanced-stage hepatoblastoma responded to chemotherapy, allowing subsequent complete hepatic resection. Both tumors had unfavorable histology. Both completed postoperative chemotherapy and have no evidence of disease (NED) with normal alpha-fetoprotein levels at 21 and 12 months, respectively, after tumor detection. One patient with stage III (group 3) bladder rhabdomyosarcoma underwent partial cystectomy following chemoradiation and is alive (NED) after 20 months. Children with BWS should be screened at regular intervals (every 3 to 6 months) for renal, adrenal, and hepatic tumors. The exact duration of screening is not yet determined.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7472930 TI - Fate of the reconstructed carotid artery after extracorporeal membrane oxygenation. AB - Reconstruction of the right common carotid artery has been shown to be feasible in neonates after extracorporeal membrane oxygenation (ECMO). However, the long term outcome after carotid artery reconstruction (CAR) remains unknown. The purpose of this study was to evaluate the natural progression of the anastomotic site after CAR. Between February 1990 and June 1993, 201 patients received ECMO. All veno-arterial (VA) ECMO patients (n = 172) were considered candidates for reconstruction unless a significant neurological event (ie, intracranial hemorrhage, stroke) had occurred; the duration of ECMO exceeded 10 days, making carotid mobilization difficult; or the patient's prognosis was deemed poor. Reconstruction was performed by excising the arteriotomy site, followed by primary end-to-end anastomosis. Reconstruction was abandoned and the artery ligated if an intimal flap, arterial thrombosis, or excessive tension was encountered. After reconstruction all patients had early carotid ultrasonography and either head computed tomography (CT) or magnetic resonance imaging (MRI). Subsequent ultrasound examinations were performed at approximately 6-month intervals. Diameter index (DI) (a measure of anastomotic narrowing) was calculated using ultrasound by dividing the anastomotic diameter by the diameter of the carotid artery 5 mm proximal to the anastomosis. Forty-three of 172 VA ECMO patients (25%) had successful reconstruction. Long-term follow-up data were available on 27 patients. These 27 patients had 39 ultrasound examinations, with an average follow-up time of 7.3 months (range, 4 days to 29 months). All carotid arteries were patent. Linear regression analysis showed significant improvement in the DI with time (P = .0001, r2 = .382).(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7472931 TI - The use of expandable metallic airway stents for tracheobronchial obstruction in children. AB - Expandable metallic angioplasty stents (Palmaz stent) have been implanted in the trachea and/or bronchi of seven children. Three children had severe tracheal stenosis after tracheoplasty for congenital tracheal stenosis repair, and four had tracheomalacia or bronchomalacia with or without vascular compression. The mean age at stenting was 9.7 months (range, 2 to 15 months). Balloon expandable stents were inserted into the trachea or bronchus through a 3.5-mm bronchoscope under fluoroscopic control. Initially a single tracheal stent was used for all patients except for one with obstruction in the trachea and both bronchi, in whom three stents were implanted. Three children had recurrent airway obstruction 1 month later; one was cured with a second stent; one child died 1 year later; and the other is being treated for heart disease. The others have no serious respiratory problems. The stents in all have been in place for 1 to 25 (mean, 11) months. No immediate complications were noted. Early and late bronchoscopy showed incomplete epithelialization of the stent and patches of granulation tissue on it. Two stents were removed bronchoscopically, one at the completion of treatment for tracheomalacia and the other at the time of recurrent airway obstruction. This preliminary experience indicates that expandable metallic stents have a useful role in the treatment of selected lower airway obstructions. PMID- 7472932 TI - Thoracoscopic closure of patent ductus arteriosus: a less traumatic and more cost effective technique. AB - The authors have developed a technique of thoracoscopic closure of patent ductus arteriosus (PDA) that significantly reduces the surgical morbidity, recovery time, and hospital costs traditionally associated with the standard open procedure. Ten patients have undergone the procedure, with nine completed successfully. One patient required conversion to an open thoracotomy. There were no operative complications, and closure of the ductus was confirmed in all cases with a postoperative echocardiogram. Eight of ten patients were discharged in under 24 hours, and hospital charges were on the average 30% to 40% less. PMID- 7472933 TI - A prospective analysis of factors influencing outcome after fundoplication. AB - Fundoplication remains a common operation in the brain-damaged pediatric patient, but recent reports suggest a poor outcome in these patients. The factors that might be associated with complications or recurrence after fundoplication have not been extensively examined. Fifty-six brain-damaged children, aged 6 months to 12 years, with documented gastroesophageal (GE) reflux underwent preoperative nutritional evaluations (percentage of ideal weight, albumin, nutrition risk index [NRI]) and documentation of medications (dexamethasone for bronchopulmonary dysplasia) before standard Nissen fundoplication. Hospital stay, intensive care unit (ICU) stay, and time on ventilator, as well as major postoperative complications (wound infection/dehiscence, pneumonia) were prospectively analyzed. Survival and recurrence rates 1 to 3 years postoperatively were also assessed. Eighty-two percent of patients were < 90% ideal weight, and 50% had NRI < 90 (normal = 100) and 29% had albumin < 3.5 g/dL. Albumin < 3.5 was significantly (P < .01) associated with prolonged hospitalization (26.8 + 2.2 versus 15.1 + 1.1 days) and ICU stay (13.8 + 1.0 versus 4.4 + .5 days) and time on ventilator (8.0 + 1.0 versus 1.8 + .4 days). NRI < 90 showed similar significant differences (P < .01). Ideal body weight < 90% was not significant. Major complications developed in 54% of patients; only two or more preoperative nutritional deficiencies, or a nutritional deficiency plus dexamethasone were significantly associated (P < .01). Recurrence occurred in 21% of patients and was significantly correlated with preoperative dexamethasone alone (P < .01), and especially when dexamethasone plus a nutritional deficit were present (low albumin, P < .001; low NRI, P < .005).(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7472934 TI - Comparison of open and laparoscopic gastrostomy and fundoplication in 120 patients. AB - The rapid development and incorporation of minimally invasive surgical techniques has abruptly changed adult surgical practices. These minimally invasive procedures are now being successfully applied to pediatric surgical problems. The anticipated benefits of these techniques include less postoperative pain, quicker return of bowel function, shorter hospital stay, and lower hospital costs, with a quicker return to normal activity. This report compares the first 60 infants and children to undergo laparoscopic gastrostomy and/or fundoplication at our institution with the same number of patients that underwent these procedures in the traditional open fashion. The two groups were similar with respect to age, sex, concurrent illness, presenting symptoms, neurological status, and procedures performed. Patients in the laparoscopic group were found to have shorter mean hospital and postoperative stays and tolerated feeding earlier. The mean hospital stay was 13.8 days for the laparoscopic group versus 16.4 days in the open group. The mean postoperative stay was 6.8 days for the laparoscopic group versus 10.7 days for the open group. The mean postoperative day on which feeding was tolerated was 2.3 in the laparoscopic group versus 4.8 in the open group. Postoperative complications were similar between the two groups. These results seem to reflect the less traumatic nature of the laparoscopic procedures as compared with the open procedures. Laparoscopic fundoplication and gastrostomy is an attractive alternative to open fundoplication and gastrostomy in infants and children. PMID- 7472935 TI - Children who are shot: a 30-year experience. AB - Three data sets describe the pattern of gunshot injuries to children from 1960 to 1993: The Harlem Hospital pediatric trauma registry (HHPTR), the northern Manhattan injury surveillance system (NMISS) a population-based study, and the National Pediatric Trauma Registry (NPTR). A small case-control study compares the characteristics of injured children with a control group. Before 1970 gunshot injuries to Harlem children were rare. In 1971 an initial rise in pediatric gunshot admissions occurred, and by 1988 pediatric gunshot injuries at Harlem Hospital had peaked at 33. Population-based data through NMISS showed that the gunshot rate for Central Harlem children 10 to 16 years of age rose from 64.6 per 100,000 in 1986 to 267.6 per 100,000 in 1987, a 400% increase. The case fatality for children admitted to Harlem Hospital (1960 to 1993) was 3%, usually because of brain injury, but the majority of deaths occurred before hospitalization. During the same period, felony drug arrests in Harlem increased by 163%. The neighboring South Bronx experienced the same increase in gunshot wound admissions and felony arrests from 1986 to 1993. The NPTR showed a similar injury pattern for other communities in the United States. In a case-control analysis. Harlem adolescents who had sustained gunshot wounds were more likely to have dropped out of school, to have lived in a household without a biological parent, to have experienced parental death, and to have known of a relative or friend who had been shot than community adolescents treated for other medical or surgical problems.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7472937 TI - Treatment of parenteral nutrition-associated cholestasis with cholecystokinin octapeptide. AB - The authors investigated whether parenteral nutrition-associated cholestasis (PNAC) in surgical neonates could be alleviated by the administration of cholecystokinin-octapeptide (CCK). Two groups of infants were studied, after major abdominal or cardiac surgery in the newborn period. The low-dose group consisted of three infants with PNAC who received cholecystokinin-octapeptide (Sincalide) at a dose of 0.02 micrograms/kg intravenously (IV), twice daily. The high-dose group comprised eight infants with PNAC who received an initial dose of 0.02 micrograms/kg IV or intramuscularly, three times daily on the first day, followed by a daily doubling of the dose up to as high as 0.32 micrograms/kg. In the low-dose group, direct bilirubin levels declined a mean of 50.2 +/- 14.5%. In the high-dose group, direct bilirubin levels declined a mean of 23.4 +/- 14.3%. In three patients in the high-dose group, no decline occurred. All three had clinical signs of overt liver failure and died of liver failure within 2 months after treatment with CCK. By excluding these patients from the high-dose group, the decline in bilirubin levels increased to 49.6 +/- 10.9%. Side effects from CCK occurred in two patients and consisted of abdominal pain and feeding intolerance. Treatment with CCK appears to be associated with a decline in direct bilirubin levels, provided overt liver failure has not developed. PMID- 7472936 TI - The Kasai procedure in the treatment of biliary atresia. AB - From 1978 to 1992, 62 patients were diagnosed as having extrahepatic biliary atresia (EHBA) at Childrens Hospital Los Angeles. The patients presented with either persistent jaundice, alcoholic stools, and/or hepatomegaly. Hepatobiliary IDA scans were performed in 47 of the patients; 46 had results typical of EHBA. Fifty-nine of the 62 patients underwent a Kasai portoenterostomy; three patients were more than 4 months of age at initial presentation and were referred directly for liver transplantation. The preoperative total bilirubin level for all patients averaged 8.6 mg/dL (range, 4.1 to 18.1). All patients underwent a standardized Kasai procedure using a 40-cm Roux-en-Y intestinal segment performed in the end-to-side fashion. Postoperative management included oral antibiotics and fat-soluble vitamins for at least 9 months. Long-term complications included cholangitis in 20 cases and portal hypertension in 25, which led to variceal hemorrhage in 12 cases. Growth rates were at or above the 50th percentile for age in 59% of the Kasai patients. Forty-one of the 59 Kasai patients survived (69.5%); six deaths occurred within 2 years after the Kasai procedure. Twelve patients were lost to follow-up within 2 years after surgery, and for calculation of mortality were presumed dead. Seventeen patients had follow-up for 5 or more years, 13 had follow-up for 2 to 5 years, and 29 had follow-up for less than 2 years. The average total bilirubin level for the patients with less than 2 years of follow-up was 7.9 mg/dL (0.3 to 20.8), and that for the patients with more than 2 years of follow-up was 1.6 mg/dL (0.3 to 18.1). Orthotopic liver transplantation was performed in 11 Kasai patients, in whom chronic liver failure eventually developed. Rejection occurred in one of these patients, which required retransplantation. Based on these results, the Kasai portoenterostomy procedure continues to offer palliation, if not long-term success, in a large percentage of patients with EHBA. PMID- 7472938 TI - Enteral feeding increases sepsis in infants with short bowel syndrome. AB - Sepsis secondary to bacterial translocation is common in infants with short bowel syndrome (SBS). Although early feeding is advocated to enhance adaptation in SBS, the effects of feeding on sepsis in SBS patients have not been examined. Twenty one infants and children (aged 2 months to 3 years) with SBS (< 80 cm small bowel length) from a variety of causes (15 necrotizing enterocolitis, 2 atresia, 2 gastroschisis, 2 volvulus) had follow-up prospectively for septic episodes before and after feedings were initiated, while still receiving total parenteral nutrition. The incidence and number of septic episodes and microbiology (blood cultures) were tabulated and compared with those of 20 patients with similar ages, and diagnoses without SBS. Statistically significant differences among infants with SBS were noted with respect to sepsis incidence (6 of 21 [29%] NPO v 16 of 21 [76%] feeding) number of septic episodes (1.3 +/- .2 NPO v 4.2 +/- .4 feeding), and presence of gram-negative rods causing bacteremia (1 of 6 [17%] NPO v 13 of 16 [81%] feeding) (all: P < .05). There were similar differences between SBS and non-SBS infants. These data show that enteral feeding increases the incidence and number of episodes of sepsis in SBS infants, but not in matched non SBS patients. The predominance of gram-negative organisms in sepsis in SBS suggests increased gut bacterial translocation in these patients, implying that selective gut decontamination may reduce the episodes of bacteremia. PMID- 7472939 TI - Preservation of postprandial intestinal hyperemic response after small bowel transplantation. AB - Postprandial intestinal hyperemia (PIH) is a normal physiological phenomenon and is thought to accommodate the increased intestinal metabolic demand during food digestion. Lack of such a response could lead to malabsorption. The purpose of this study was to determine whether transplanted intestine retains the ability to mediate PIH. Nineteen rats with syngeneic orthotopic small bowel transplantation (SBT) and 13 normal controls were further divided into two subgroups to receive gastric gavage of either 2 mL of Ensure Plus (EP) high-calorie liquid nutrition or 2 mL of normal saline. Intestinal blood flow was determined before, and 20 and 45 minutes after the gavage, employing a colored microsphere technique. The results showed that basal blood flow to the transplanted intestine (56.7 +/- 26.7 mL/min/100 g) was similar to that of normal intestine (64.2 +/- 35.8 mL/min/100 g; P > .1). EP feeding elicited a marked increase in intestinal blood flow (PIH phenomenon) in both normal and SBT animals. The magnitudes of flow increase were similar for the two groups (249 +/- 99% versus 241 +/- 76% of baseline 45 minutes after EP feeding in the normal and transplanted intestine, respectively; P > .1). The results indicate that hyperemic response to feeding is normal in syngeneic orthotopic SBT. PMID- 7472940 TI - Prevention of fetal bowel allograft rejection by combined treatment with anti ICAM-1 and anti-LFA-1 antibodies. AB - Prevention and treatment of allograft rejection remain the major issues in clinical small bowel transplantation. New strategies for manipulating immune responses using more powerful immunosuppressive agents continue to be evaluated. The fetal small bowel from BALB/c (H-2d) or C3H/He (H-2k) mice was transplanted into the space between the peritoneum and rectus abdominis of adult C3H/He (H-2k) recipient mice. Syngeneic (n = 6) and allogeneic transplant groups were made. In the allogeneic group, the recipient mice were subdivided into three groups, depending on the duration of combined treatment with anti-LFA-1 and anti-ICAM-1 monoclonal antibodies (MAbs): untreated (n = 10), 7-day course (n = 10), and 4 week course (n = 14). A dose of 50 micrograms/mouse/d each of both MAbs was given intraperitoneally, immediately after transplantation and on the consecutive days. All mice were killed 4 weeks after transplantation, and the graft as well as the recipient spleen were taken for histological examination, graft survival ratio, mixed lymphocyte reaction (MLR) assay and cytotoxic T lymphocyte (CTL) assay. All grafts in the syngeneic group survived with normal villi, whereas all grafts in the allogeneic group without treatment disappeared within 4 weeks. All grafts in the allogeneic group with a 7-day course of MAb treatment showed marked disruption of the mucosa with massive cellular infiltration. However, in the allogeneic group treated for 4 weeks, all allografts had adequate growth and demonstrated normal villi with minimal cellular infiltration. Splenocytes from allografted recipient mice without MAb treatment showed markedly increased MLR and CTL activity, compared with the activity seen in the syngeneic MLR and CTL.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7472941 TI - Successful prenatal management of hydrops, caused by congenital cystic adenomatoid malformation, using serial aspirations. AB - Congenital cystic adenomatoid malformation (CCAM) is a lung lesion that is now commonly diagnosed in utero with fetal ultrasonography. The described treatment of this lesion includes observation with treatment delivery, a single aspiration, thoraco-amniotic shunts, and fetal resection. This patient had an in utero diagnosis of a Stocker type I CCAM associated with hydrops. Fetal resection was not an option because of patient refusal. The fetus was treated with multiple serial aspirations. There was marked improvement of the anasarca, and subsequently the baby was born without respiratory distress. On the second day of life the CCAM began to expand, and the right lower lobe was resected. The baby's postoperative course was uneventful. CCAM with hydrops is associated with a high mortality rate. Current recommended therapy for these lesions is fetal resection or thoracoamniotic shunt. The authors' patient was treated with serial fetal thoracocenteses, with an excellent outcome. This therapy may be an alternative to fetal surgery or an adjunct to fetal surgery in selected cases. PMID- 7472942 TI - Aldosteronoma in childhood. PMID- 7472943 TI - Surgical and critical care management of children with life-threatening injuries: the Swiss experience. PMID- 7472944 TI - Pediatric surgery between 1860 and 1900. AB - The twentieth century begins the modern era of pediatric surgery, a time of innovation and vastly improved operative survival. Until then, many correctable congenital anomalies had been described in case reports only as medical curiosities, seemingly having no chance for surgical cure, but the challenge was available to those wishing to accept. The first texts devoted to surgical diseases of infancy and childhood were descriptive, emphasizing visible and palpable lesions, tuberculosis, musculoskeletal deformity, and major trauma, but also recognized that "surgical diseases of children by their number, their variety, and their character must, we think, be studied in a special manner and demand a special chapter in surgical books." The construction of Children's Hospitals during the nineteenth century identified the need to provide special facilities for children. Surgical antisepsis was practiced and general anesthesia used. Physicians such as Hirschsprung recognized a wide spectrum of treatable surgical conditions that would soon become the domain of pediatric surgeons. Indeed, at that moment in time at the very beginning of the twentieth century, Roentgen discovered the x-ray, Ladd received his MD degree at Harvard, the first pediatric appendectomies for appendicitis were successfully performed, and Fredet and Ramstedt recognized that pyloroplasty was not the preferred treatment for hypertrophic pyloric stenosis. Modern pediatric surgery had passed from embryo to fetus to newborn. PMID- 7472945 TI - Experimental fetal tracheal ligation and congenital diaphragmatic hernia: a pulmonary vascular morphometric analysis. AB - The authors have previously shown that fetal tracheal ligation (TL) reverses the pulmonary hypoplasia in experimental diaphragmatic hernia (DH) by accelerating fetal alveolar growth. The purpose of this study was to determine if growth of the accompanying macroscopic and microscopic pulmonary vasculature is also accelerated. Eighteen fetal lambs were divided into three experimental groups: diaphragmatic hernia (DH), DH and simultaneous tracheal ligation (DH/TL), and sham-operated controls (C). Animals were delivered near term, the lungs retrieved, and pulmonary capillary growth (5 to 50 microns in diameter) evaluated by standard morphometric techniques. Capillary ultrastructure was evaluated by electron microscopy. Nine additional fetal lambs of the same gestational age were equally divided into the same three groups and their lungs analyzed by pulmonary arteriography for evaluation of large vessel growth (< 100-microns diameter). Computer digital analysis of angiogram lung slices showed that the total area of large vessels was increased in DH/TL lungs when compared with DH lungs and decreased in DH lungs when compared with C lungs (P = .003); however, the ratio of large vessel area per unit of lung area was similar in all groups. Microscopic morphometry of the capillary bed showed that the total number of capillaries was increased in DH/TL lungs over both DH and C lungs (P = .0001); however, the number of capillaries per alveolus (cap/alv) was similar in all groups. In DH/TL lungs, electron microscopy showed normal capillary wall structure and normal thickness of the capillary-alveolar interface, whereas in DH lungs, capillary structure was abnormal and the capillary-alveolar interface was thickened.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7472946 TI - Pathophysiology of congenital diaphragmatic hernia. XI: Anatomic and biochemical characterization of the heart in the fetal lamb CDH model. AB - AIM: The purpose of this study was to determine whether the presence of bowel in the chest during development in the fetal lamb model of congenital diaphragmatic hernia (CDH) results in structural and/or biochemical hypoplasia of the left venticle. METHODS: The model was created at 80 days' gestation and delivered at term. The hearts were fixed in 4% formaldehyde solution, components weighed, and right ventricular (RV) and left ventricular (LV) wall thicknesses and both aortic (Ao) and pulmonary artery (PA) root diameters were measured. Fresh specimens were analyzed for protein, DNA, hydroxyproline, and elastin content. All CDH measurements are compared with littermate control tissues. RESULTS: There were no differences in body weight (kg) between CDH and control littermates (4.25 +/- 0.26 versus 3.71 +/- 0.24, P = NS). CDH lambs have significantly decreased total heart (4.88 +/- .25 versus 6.75 +/- .49, P < .05), left ventricular (1.65 +/- .11 versus 2.15 +/- .19, P < .05), septal (1.29 +/- .11 versus 1.99 +/- .21, P < .05), and combined atrial (0.68 +/- .06 versus 1.14 +/- .15, P < .05) weights (g/kg lamb) without differences in RV weights (1.26 +/- .07 versus 1.57 +/- .17, P = NS). LV and RV wall thickness, and Ao root diameters (cm) were found to be identical in both CDH and control lambs. However, PA root diameters (0.47 +/- .01 versus 0.38 +/- .01, P < .005) and ductus arteriosus diameters were increased in CDH (0.35 +/- .01 versus 0.22 +/- .02, P < .005). Total protein, DNA collagen, and elastin content and DNA/total protein ratios were identical in RV and LV in both CDH and control lambs. CONCLUSION: Newborn lambs with left-sided CDH have a significantly lower total heart, LV, septal, and atrial weights without differences of RV weight or ventricular wall thicknesses. Given these findings, the unchanged DNA/protein ratio implies that the left ventricle is hypoplastic in CDH. Ao/PA root ratios suggest that LV hypoplasia in utero may result in increased left atrial pressures, decreased right-to-left shunting through the foramen ovale, and increased PA pressures and flow, resulting in increased PA root and ductus arteriosus diameters. This model simulates the clinical data from human fetuses/neonates with CDH. Further investigations are necessary to determine the functional significance of these findings. PMID- 7472947 TI - Severity of hypoxia predicts response to nitric oxide in a porcine pulmonary hypertension model. AB - Although inhaled nitric oxide (NO) has been variably successful in resolving pulmonary hypertension in neonates, children, and adults, no parameters predictive of response to this therapy have been elucidated. We conducted an animal study to determine if severity of hypoxia can predict magnitude and sustenance of response to inhaled NO therapy. Seven Yorkshire swine weighing 11 to 20 kg underwent 16 experiments, each consisting of four phases: Phase 1: Control period of ventilation on FIO2 .3; phase 2: Hypoxic period on FIO2 .10 to .15, establishing pulmonary hypertension; phase 3: Treatment period with NO starting at five parts per million (ppm), doubling dose every 10 min to 80 ppm; phase 4: Posttreatment observation period after discontinuation of NO while maintaining hypoxia for 1 hour or until circulatory failure or pulmonary hypertension of pre-NO magnitude developed. Each animal underwent a maximum of three experiments in random order of hypoxia severity before sacrifice with pentobarbital overdose. Continuous hemodynamic parameters, intermittent cardiac output and pulmonary capillary wedge pressure, and intermittent arterial blood gas analyses were obtained through pulmonary and systemic artery catheters placed by femoral cutdown. Pulmonary and systemic vascular resistances (PVR and SVR) were calculated by standard formulas. Experiments were divided into two groups (n = 8 in each): group 1 with severe hypoxia (PaO2, 25 to 35) and group 2 with moderate hypoxia (PaO2, 36 to 65). Data for all hemodynamic parameters were expressed as mean percentage change from baseline (phase 1) +/- SEM under each set of conditions, and the two groups were compared by two-way analysis of variance and covariance adjusted for order of experimentation.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7472948 TI - Infants with metastatic neuroblastoma have improved survival with resection of the primary tumor. AB - Although infants (age less than 1 year) with neuroblastoma have a favorable overall prognosis, metastatic disease is associated with poorer treatment outcome. To assess the role of surgery in these patients, the authors reviewed survival data for 151 infants treated for neuroblastoma, focusing on patient and tumor characteristics, biological markers, and surgical management among the 99 patients with metastatic disease. Patients were divided into early (1961 to 1978) and contemporary (1979 to 1993) treatment eras. Potential prognostic factors were statistically tested to determine their significance in affecting survival. Five year survival by Pediatric Oncology Group stage was: A, 100% (+/- 0%); B, 94% (+/ 6%); DS, 77% (+/- 9%); C, 73% (+/- 9%); and D, 61% (+/- 8%). Survival for infants with metastatic disease (stages C, D, and DS) was affected significantly by treatment era (P = .0001). Analyses restricted to patients treated during the contemporary era showed prognostic significance for DNA index (P = .02), N-myc copy number (P = .007), serum lactate dehydrogenase level (P = .001), and extent of resection (P = .01). A > or = 95% resection of the primary tumor was found to be associated with improved survival. Significantly more surgical complications were associated with resections performed at the time of diagnosis (P = .007), and delaying surgery until after several courses of chemotherapy did not decrease survival. In conclusion, multiple factors affect the outcome of treatment for infants with metastatic neuroblastoma, and whenever feasible, a > or = 95% resection of the primary tumor should be performed in this patient subgroup. PMID- 7472949 TI - Conservative management of uterine pediatric rhabdomyosarcoma: a report from the Intergroup Rhabdomyosarcoma Study III and IV pilot. AB - Previous studies have suggested that women with uterine rhabdomyosarcomas (RMS) represent a distinct group of patients who present at an older age, are less responsive to treatment, and have a poorer prognosis than patients with vaginal RMS. During the Intergroup Rhabdomyosarcoma Study (IRS) III and the IRS IV pilot study, 14 patients were registered with uterine primary RMS. Three patients presented with cervical tumors that were completely removed (group 1). Eight patients had initial biopsies with gross residual disease (group 3), and 3 had metastatic disease at presentation (group 4). Of the 5 patients treated with primary chemotherapy or chemotherapy and radiation, 2 had delayed hysterectomy and vaginectomy, 1 had no further surgery, and 2 had exploratory laparotomy with no evidence of disease. There were no relapses or deaths in this group. One patient underwent initial resection of a broad ligament mass, experienced an early (3-week) recurrence of the mass while on chemotherapy, and progressed to developing distant metastases and death. Four patients died of chemotherapy toxicity or sepsis, one after achieving a complete response from chemotherapy and hysterectomy. This primary chemotherapy or chemotherapy and radiotherapy regimen resulted in 8 of 9 (89%) patients (not including those who died of chemotoxicity) surviving between 1.5 and 6 years without evidence of disease. Of the surviving patients, 2 had hysterectomy and vaginectomy, but pathological specimens showed only localized microscopic residual tumor. This report suggests that less vigorous operative resection may be possible in combination with primary chemotherapy when treating uterine rhabdomyosarcomas.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7472950 TI - Regenerative healing of incisional wounds in murine fetal lungs maintained in organ culture. AB - Although fetal dermal repair is known to be fundamentally different from adult healing, the response to wounding in other organs is less well characterized. Scarless repair in mid-gestation dermis with a transition to adult-type healing at term has been shown in fetal organ culture. A lung explant culture system was used to investigate whether wound repair in the fetal lung shows characteristics similar to those found in fetal dermis. Lungs from 14-day and 18-day Cd-1 murine fetuses and 2-day-old newborns, (term = 20 days, n = 24) were wounded by linear incision and incubated at 37 degrees C, in a 21% O2, 5% CO2 environment, in BGJb supplemented with vitamin C and antibiotics. Medium was changed daily. Samples were fixed at 7 days and embedded in paraffin. Sections were stained with hematoxalyn-eosin and Masson Trichrome. Additional 14-day and 18-day samples were frozen in freon and immunohistochemical staining for TGF-beta performed. Other frozen tissues from each time point were homogenized and used to assay for endogenous TGF-beta levels by Western blot analysis. Histology showed reconstitution of tissue architecture across the wound in 14-day and 18-day specimens. In representative histological sections, intact bronchial architecture developed across the previous wound site. No cellular inflammatory response was observed, and collagen deposition was undetectable at the site of the wound by Trichrome staining. By 22 days the lung explants showed a much less ordered repair, including disorganized collagen deposition.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7472951 TI - Hyaluronic acid metabolism in keloid fibroblasts. AB - Hyaluronic acid (HA), an important component of the tissue extracellular matrix, is a ubiquitous glycosaminoglycan (GAG) that forms a pericellular coat on the surface of cells. It has been speculated that this pericellular HA boundary may localize cytokines, such as transforming growth factor-beta 1, which is known to stimulate collagen production. The purpose of this study was to examine the role of HA and its cell surface receptor (CD44), an active participant in HA degradation, as they relate to keloid formation. Dermal excisions from both normal patients (n = 13) and keloid patients (n = 13) were analyzed for HA content using an alcian blue staining technique. Fibroblast cell cultures were used to quantitate HA synthesis and CD44 receptor density. Histological analyses showed a greater HA content in keloid tissue compared with normal dermal tissue. In agreement with this observation, keloid fibroblasts were found to synthesize significantly more HA than normal dermal fibroblasts (2469 +/- 483 cpm versus 1122 +/- 256 cpm, P = .02). Treatment of keloid fibroblasts with triamcinolone acetonide reduced the level of HA synthesis to that of normal fibroblasts (1560 +/- 477 cpm versus 1293 +/- 264 cpm, P = .6). However, there was no significant difference in HA receptor density on keloid cells compared with normal skin fibroblasts. Therefore, the increased HA deposits found in keloids are attributable to increased synthesis rather than to decreased degradation mediated by the CD44 receptor. PMID- 7472952 TI - Decreased cysteine and proline synthesis in parenterally fed, premature infants. AB - Little is known about the amino acid (AA) biosynthetic capacity and requirements of premature infants. This study assessed the synthesis of seven biochemically nonessential AA from a universal precursor, glucose, in stable, parenterally fed, premature neonates. Seven infants (six boys, one girl) were studied at a mean age of 6.3 +/- 0.6 (SEM) days; mean gestational age was 29.7 +/- 1.3 (SEM) weeks, and mean birth weight was 1,222.8 +/- 176.5 (SEM) grams. All infants were parenterally fed a mixture of 7.5% to 12.5% dextrose and 2.2% Trophamine, with or without lipid. Mean caloric intake was 93 +/- 8.4 (SEM) kcal/kg/d, and total AA intake was standardized at 2.86 g/kg/d AA, plus supplemental cysteine (30 mg/g AA/d). Each infant received a 4-hour continuous, unprimed intravenous infusion of a stable isotope tracer of D(-)[U13C] glucose (200 mg/kg). Blood samples were obtained before and at the end of the infusion. Conversion of the glucose tracer into seven biochemically nonessential AA (cysteine [Cys], proline [Pro], aspartate [Asp], serine [Ser], glutamate [Glu], alanine [Ala], and glycine [Gly]) was assessed by measuring their isotopic enrichment in plasma, using gas chromatography/mass spectrometry (GC/MS), and expressed as mole percent excess (MPE) (mean +/- SEM). The isotopic enrichment of plasma glucose was also measured using GC/MS. Free plasma AA concentrations (mean +/- SD) were measured using an automated amino acid analyzer. Mean MPE for M + 1, M + 2 and M + 3 Cys, and for M + 1 and M + 3 Pro were not significantly different from 0; M + 2 Pro barely achieved statistical significance (P = .048).(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7472953 TI - Weanling and adult rats differ in fatty acid and carnitine metabolism during sepsis. AB - Increased oxidation of fat is an important host response to sepsis, and carnitine is essential for long-chain fatty acid oxidation. Because neonates have low levels of carnitine, their ability to respond to a septic insult may be impaired. The purpose of this study was to compare fatty acid and carnitine metabolism in septic weanling (60 to 85 g) and septic adult (285 to 310 g) rats. Sepsis was induced in weanling and adult male Sprague-Dawley rats by cecal ligation and puncture (CLP). The rats were killed 16 hours after CLP or sham operation, and serum glucose, lactate, beta-hydroxybutyrate, fatty acid, carnitine, liver fatty acid, and tissue carnitine levels were measured. The data suggest that during sepsis weanling rats may be more dependent on fatty acid oxidation than adult rats are, as evidenced by their elevated serum fatty acid and acylcarnitine levels, and relative hypoglycemia and hyperketonemia. In addition, although total serum carnitine levels were increased in both adult and weanling septic rats, tissue carnitine levels of weanling rats became significantly depleted during sepsis, unlike in adult rats. This study supports further investigation regarding the role of exogenous carnitine in newborn sepsis. PMID- 7472954 TI - Octreotide does not prevent bacterial translocation in an infant piglet model of intestinal ischemia-reperfusion. AB - The process of bacterial translocation (BT) after ischemia/reperfusion (I/R) injury is reported to be mediated by local mucosal factors, the effects of pancreatic enzymes, epithelial disruption, and by dysfunctional intestinal motility. Octreotide (OCT), a somatostatin analog, has been postulated to protect against BT by influencing one or more of these factors. Twenty-two formula-fed piglets (weight, 3.5 +/- 0.5 kg; age, 20 +/- 5 days) were divided into four groups: control (no drug given; no I/R; n = 6), I/R (no drug given; n = 5), I/R plus low-dose OCT (LD OCT, 0.08 microgram/kg; n = 6), and I/R plus high-dose OCT (HD OCT, 8 micrograms/kg; n = 5). All experimental subjects had nonocclusive mesenteric ischemia induced by reversible pericardial tamponade with mesenteric flow decreased to 25 +/- 5% of baseline for 5 hours followed by 15 +/- 5 hours of reperfusion. Mesenteric lymph nodes (MLN), liver, spleen, blood, and peritoneum were harvested for blind microbial analysis. None of the animals in the control group experienced translocation to the tissues tested. All of the animals in the I/R group experienced BT to the MLN. The subjects in the LD OCT and HD OCT groups experienced BT to the MLN 66% and 80% of the time, respectively. Despite the reported clinical evidence that OCT can protect the intestinal mucosa from injury and increase the clearance of bacteria from the gastrointestinal tract, in this study in which variables other than I/R known to promote bacterial translocation were eliminated, OCT failed to modify or prevent the occurrence of translocation to the MLN after I/R injury. PMID- 7472955 TI - Laparoscopic cholecystectomy versus open cholecystectomy in children: which is better? AB - Twenty-nine consecutive laparoscopic cholecystectomies (LC) performed between April 1992 and December 1993 were compared with 23 consecutive open cholecystectomies (OC) performed between January 1991 and March 1992 with regard to clinical, surgical, and economic factors. Most patients were Caucasian (> 70%), and symptomatic nonhemolytic cholelithiasis was the most common indication for cholecystectomy. The introduction of LC did not significantly increase the number of cholecystectomies performed per annum. There is a learning curve to LC: the average length of operative time required during the first year was significantly longer than that of OC and the average time for LC during the second year (P < .01). By the second year, the average operative time of LC was not significantly different from OC. There was no conversion from LC to OC, and the complication rate was minor in both groups. The postoperative parenteral analgesic requirement for LC was significantly less than OC (P < .01). The mean length of hospitalization of LC was about three times shorter than that of OC (P < .01). Although the average operating cost per case of LC was significantly more expensive than OC, LC was significantly cheaper because the period of hospitalization was significantly shortened (P < .01). In conclusion, LC is the procedure of choice in the treatment of symptomatic cholelithiasis in children. PMID- 7472956 TI - Amniotic band syndrome in fetal lambs. I: Fetoscopic release and morphometric outcome. AB - A fetal lamb model of amniotic band syndrome (ABS) was developed to study the pathophysiology of banded extremities and evaluate the possibility of in utero treatment with the potential for functional recovery. Eight fetal lambs underwent banding of their extremities with umbilical tape at 100 days' gestation. Two lambs aborted after the open fetal surgery. The limbs of two unoperated newborn lambs served as controls in addition to five sham-operated control limbs that had no bands applied. Nine limbs were banded without reducing blood flow assessed by laser doppler (group 1), and 6 limbs were similarly banded and released fetoscopically at 125 days' gestation. Four limbs were banded, with a mean reduction in blood flow to the limb of 18.7% (group 2) by laser doppler flowmeter. Shortly after birth the lambs were killed, and segmental limb length, circumference, joint range of motion, and histology were evaluated. At birth, banded limbs showed marked brawny edema and absence of wool distal to the band. Segmental limb measurements showed shorter distal forelimbs in banded limbs compared with controls (10.97 +/- 0.59 versus 12.98 +/- 0.69, P < .05). Banded limbs were associated with progressive increase in hoof circumference (P < .03) and a decrease in joint range of motion (P < .003). In sharp contrast, there were no differences between fetoscopically released limbs and control limbs in any of the parameters measured. Histology of banded extremities showed edema, venous and lymphatic congestion, and fibrosis compared with controls. This model of ABS in fetal lambs is simple, reproducible, and replicates all the clinical features of extremity ABS.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7472957 TI - The natural history of meconium peritonitis diagnosed in utero. AB - The authors reviewed their experience with meconium peritonitis (MP) diagnosed in utero to define criteria for prenatal and postnatal management. Prenatal diagnosis was made by identifying abdominal calcification on serial ultrasound examinations in nine fetuses, between 18 and 37 weeks' gestation. Cases without associated bowel abnormalities were considered "simple MP" and those with bowel abnormalities were considered "complex MP." Five cases of simple MP were identified at 18, 23, 30, 34, and 37 weeks' gestation. These five fetuses were delivered at term and had normal abdominal examinations. Abdominal radiographs were obtained in three showing normal bowel gas patterns, and abdominal calcifications in only two. All five patients were fed uneventfully. Four cases of complex MP were identified at 26, 26, 31, and 31 weeks' gestation. All four fetuses had dilated loops of bowel. Two of the four had meconium cysts, one of which was associated with ascites and the other with polyhydramnios. Shortly after birth both infants with meconium cysts required ileal resection and ileostomy for ileal atresia and ileal perforation, respectively. The remaining two infants had no evidence of dilated bowel, meconium cyst, or ascites on postnatal radiograph and were fed uneventfully. These data suggest that only 22% of fetuses with a prenatal diagnosis of MP develop complications that require postnatal operation. Gestational age at diagnosis does not correlate with postnatal outcome. Fetuses with complex MP are at increased risk for postnatal bowel obstruction and perforation. PMID- 7472958 TI - Effect of intraamniotic dexamethasone administration on intestinal absorption in a rabbit gastroschisis model. AB - Infants with gastroschisis experience delayed intestinal motility and absorption for several weeks after birth. This intestinal dysfunction is believed to occur primarily in the third trimester and to be largely caused by the prolonged exposure of the intestine to amniotic fluid. Previous studies have shown that prenatal steroid administration will enhance mucosal disaccharidase activity and nutrient uptake. The present study evaluates the effects of dexamethasone on intestinal function in a rabbit fetal gastroschisis model. Thirty-four fetuses from 10 New Zealand white rabbits were divided into three groups: (1) gastroschisis group (GSC, n = 10), gastroschisis was created on gestational day (GD) 24 (term = 31 to 33 days); (2) dexamethasone group (GSD, n = 10), after the creation of gastroschisis, a small osmotic pump was placed into the rabbit doe for dexamethasone infusion into the fetal amniotic cavity for 7 days (0.2 microgram/g/d); (3) normal group (NF, n = 10), unoperated littermates from the GSC group. There were no maternal deaths, and fetal survival rate was 85%. The fetal small intestinal disaccharidase enzyme, lactase (UE/g protein), was markedly decreased in GSC fetuses. It was increased 70% in the GSD group but lower than in normal fetuses (GSC = 10.0 +/- 1.6; GSD = 17.3 +/- 1.6 [GSD versus GSC, P < .05]; NF = 48.0 +/- 6.7). Maltase activity in the GSD group was significantly increased (GSC = 7.2 +/- 1.1; GSD = 13.9 +/- 1.8 [GSD versus GSC, P < .05]; NF = 12.2 +/- 1.3).(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7472959 TI - Early postoperative alterations in infant energy use increase the risk of overfeeding. AB - AIM OF STUDY: Energy needs in infants are decreased after surgery because of growth inhibition (resulting from catabolic stress metabolism), decreased insensible losses, and inactivity. Using standardized formulas that account for growth, activity, and insensible losses during this stress period can lead to overfeeding in excess of 200% of the actual measured requirement. Overfeeding during this acute injury period can result in increased CO2 production from lipogenesis. This study determined the effects of a reduced rate of mixed caloric repletion on infant energy use during the early postoperative period. METHODS: C reactive protein (CRP), oxygen consumption (VO2), carbon dioxide production (VCO2), measured energy expenditure (MEE), and total urinary nitrogen (TUN) were measured serially in seven infants (average age, 78 days) during the first 72 hours after abdominal or thoracic surgery. Nonprotein respiratory quotient (RQnp), and values for oxidation of carbohydrate (Ce) and fat (Fe) were calculated. Injury severity was stratified based on serum CRP concentrations of > or = 6.0 mg/dL (high stress) or < 6.0 mg/dL (low stress). Recovery from acute stress was analyzed by comparing studies in which CRP had decreased to < or = 2.0 mg/dL (resolving stress group) with those in which CRP values were greater than 2.0 mg/dL (acute stress group). RESULTS: Average total caloric intake (64.56 +/- 18.51 kcal/kg/d; approximately 50% of predicted energy requirement) exceeded average MEE (42.90 +/- 9.98 kcal/kg/d) by approximately 50%. Average TUN was 0.18 +/- 0.07 g/kg/d (high stress 0.2 +/- 0.05 versus low stress 0.16 +/- 0.09 g/kg/d). Average RQnp was 1.05 +/- 0.13 and average Ce was 37.28 +/- 16.86 kcal/kg/d. The average calculated Fe was 0.0 +/- 12.27 kcal/kg/d, reflecting approximately equal amounts of fat oxidized compared with fat generated from excess glucose (lipogenesis). When individual studies were analyzed at a CRP cutpoint of 2.0 mg/dL, overfeeding (RQ > 1.0) was significantly less likely in the resolving (2/6 studies, 33.4%) versus acute stress (9/13 studies, 69.2%, Z test P < .001) group. Five of seven (5/7) patients (9/19 individual studies) had negative Fe values (average -9.89 +/- 10.02) reflecting net lipogenesis. The RQnp for these nine studies was 1.14 +/- 0.11 versus 0.97 +/- 0.09 for the remaining 10, and this difference was significant (P < .01). A significant correlation existed between carbohydrate intake and VCO2 (Pearson r = .6951, P < .01). In addition, there was a good correlation between carbohydrate intake and VCO2 (Pearson r = .6591, P < .01). The coefficient of variation for MEE was 8.0% (low stress) versus 30.2% (high stress). CONCLUSION: Lipogenesis with increased CO2 production is substantial, even at reduced caloric delivery rates that exceeded MEE by only 50%, during the early postoperative acute metabolic stress period in infants. These data suggest that caloric requirements during stress are likely equal to or only minimally in excess of actual MEE. Intersubject variability, especially in more severely stressed infants, underscores the importance of serial measurements of energy expenditure to enable precise caloric delivery and avoid overfeeding. In the absence of calorimetric measurement, the data suggest that PBMR (predicted basal metabolic rate) should be used to estimate caloric delivery until CRP values are < or = 2.0 mg/dL. PMID- 7472961 TI - Selective distal splenorenal shunts for intractable variceal bleeding in pediatric portal hypertension. AB - The treatment of portal hypertension in the pediatric population has undergone an evolution toward less invasive methods of care. With the advent of endoscopic sclerotherapy, surgery is less common in the acute care of these patients. Few reports deal with the role of portosystemic shunting in the emergent management of variceal hemorrhage in children. To address this issue, the authors studied the medical records of all pediatric patients at their institution who underwent placement of a shunt for portal hypertension during the last 10 years. Nine patients underwent a total of 10 emergent or semiurgent shunting procedures. Seven were boys and two were girls. Six patients had portal hypertension as a result of intrahepatic disease. Two had extrahepatic portal vein thrombosis. Five children had abnormal hepatic function. The median age at the time of the procedure was 9 years. The indication for surgical shunting in all cases was gastrointestinal hemorrhage not responsive to sclerotherapy. Eight patients underwent emergent distal splenorenal shunts (DSRS), and two underwent a nonselective mesocaval shunt, with one undergoing both. Postoperatively all patients had cessation of bleeding. Operative mortality was zero. Early complications included ascites (3), small bowel obstruction (1), and hepatorenal syndrome (1). The child who underwent a nonselective shunt procedure had encephalopathy. Two DSRS thrombosed, requiring reexploration; eight shunts remained patent. Three patients eventually had orthotopic liver transplantation (OLT) because of progressive hepatic failure. Two children died; neither death was shunt related.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7472962 TI - Persistent hyperinsulinemic hypoglycemia of infancy: experience with 28 cases. AB - Twenty-eight infants with persistent hyperinsulinemic hypoglycemia of infancy (PHHI) were seen during a 10-year period. There were 13 males and 15 females. Their age at time of presentation ranged from a few hours to 6 months. Consanguinity was reported in 20 cases (71.4%). One family had two affected siblings and two affected cousins, another had three affected siblings and one affected cousin, and three others had lost siblings because of hypoglycemia and seizures. The primary clinical presentation was jitters and seizures in association with hypoglycemia. The diagnosis was suspected when the therapeutic glucose requirement was found to be more than 12 mg/kg/min and also when there was a good response to glucagon after exclusion of metabolic and storage diseases. A high insulin-to-glucose ratio was noted for all patients. Twenty-two had near-total (90%) pancreatectomy; the result was excellent in all but four, who required supplemental medical therapy. Five patients were treated medically, and one patient's family refused treatment. Twelve patients sustained moderate to severe brain injury before referral. There were no deaths, and only one patient had evidence of malabsorption after the pancreatectomy. PHHI correlates well with consanguinity and family history. Clinical awareness is essential to permit early diagnosis and prompt medical and supportive therapy.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7472960 TI - Complications after surgical intervention for necrotizing enterocolitis: a multicenter review. AB - Necrotizing enterocolitis (NEC) is a serious condition affecting predominantly the premature infant. The purpose of this study is to report a multicenter experience of complications in 252 infants requiring surgical therapy for NEC. Data from eight institutions for the years 1980 through 1990 were collected and analyzed for infants undergoing surgical therapy for NEC. Records were reviewed for gestational age, birth weight, age at operation, indications for operation, degree of intestinal involvement, operation(s) performed, complications, and 30 day mortality rates. A total of 264 infants underwent surgical intervention for NEC during the study period. Complete information was available for 252 patients. The mean gestational age was 31 +/- 5 weeks and the mean birth weight was 1,552 +/- 823 g. The mean age at operation was 18 +/- 35 days. Pneumoperitoneum was the most common indication for operation (42%). The 30-day survival rate was 72%. Eighty-one percent of patients underwent primary laparotomy, whereas peritoneal drainage was performed in 48 (19%) patients. Postoperative complications were identified in 119 (47%) patients. The most common postoperative complications were sepsis (9%), intestinal strictures (9%), and short gut (9%). Wound infections occurred in 6%, and the incidence of intraabdominal abscess formation was only 2.3%. Gestational age < 27 weeks (P < .005) and birth weight < 1,000 g (P < .005) were associated with significantly increased mortality but no increase in postoperative morbidity. The incidence of complications was similar in the very low birth weight (< 1,000 g) infants (51%) compared with infants > or = 1,000 g (46%).(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7472963 TI - Vascular complications after pediatric liver transplantation. AB - From February 1986 to July 1994, 81 hepatic transplantations were performed in 73 children, with an overall patient survival rate of 83%. Forty-two patients received whole-liver grafts (WLG) and 39 had reduced-size grafts (RSG). The mean patient weight was 19.7 kg, with 29 patients weighing less than 10 kg. Seventeen vascular complications (21%) occurred in 13 children: 8 (10%) had hepatic artery thrombosis (HAT), 5 (6%) had portal vein thrombosis (PVT), 1 had both HAT and PVT (1%), and 3 (4%) had aortic conduit perforation (ACP). There was no significant difference in the incidence of HAT between RSG (5%) and WLG (14%) or between children weighing less than 10 kg (10%) and those weighing more than 10 kg (10%). The site of arterial reconstruction, end-to-end to the recipient common hepatic artery or end-to-side to the infrarenal aorta, had no significant effect on the occurrence of HAT (7% v 8%), but HAT occurred in 2 of 6 cases (33%) in which an aortic conduit was used. PVT documented in 5 cases (6%) was associated with technical complications (2), preduodenal portal vein (2), and a circulating cardiolipid antibody (1), and required thrombectomy, with no graft loss. Combined HAT and PVT was found in one patient 2 years postretransplantation for HAT. Although graft function is normal, portal hypertension persists. The aortic conduit, used in six patients, led to arterial perforation (3), HAT (2), and death (2). Of the 8 cases of HAT, 1 was diagnosed during autopsy and 7 occurred within 30 days and required retransplantation (6) or thrombectomy with rearterialization (1).(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7472964 TI - Hepatic artery ligation for hepatic vascular tumors with arteriovenous and arterioportal venous shunts in the newborn: successful management of two cases and review of the literature. AB - The authors report their experience with hepatic artery ligation in two newborns, as well as relevant findings from a literature review. A 2-day-old boy had progressive cardiac and respiratory difficulty. A firm liver was palpable, with an overlying thrill. Sonography and arteriography showed diffuse arteriovenous shunting in both liver lobes. Hepatic artery ligation provided remarkable hemodynamic and clinical improvement. Another boy was admitted 3 weeks after birth because of bilious vomiting with abdominal distension and bloody stools. Abdominal examination showed a large liver with a systolic bruit and thrill. X rays showed cardiac enlargement and dilated bowel loops with air-fluid levels. Arteriography and sonography showed arteriovenous and arterioportal venous shunting. Laparotomy was performed, and a large vascular malformation was palpated in both liver lobes. The entire bowel was congested and cyanotic, but there were no signs of obstruction. This patient had acute portal hypertension imitating intestinal obstruction. Ligation of the hepatic artery improved the color of the bowel, and the thrill disappeared. Five and nearly 4 years after the operation, both boys are growing normally without medication or diet. Sonography showed almost complete resolution of the hemangiomas. PMID- 7472965 TI - Necrotizing fasciitis in childhood. AB - Necrotizing fasciitis is a rare entity in the pediatric population. Five cases of this soft tissue infection were treated at the authors' institution between January and December 1993. Three of the children were profoundly neutropenic secondary to chemotherapy. All five were treated with aggressive surgical debridement, frequent dressing changes, broad-spectrum antibiotics, and nutritional support. In addition, the patients with neutropenia received a combination of granulocyte-colony stimulating factor and granulocyte transfusions. One child died of overwhelming sepsis and bone marrow graft failure. The others eventually made a complete recovery. Necrotizing fasciitis may be becoming a more common problem in children. Aggressive chemotherapeutic regimens and more frequent use of bone marrow transplantation could be a factor in this. Early diagnosis and aggressive surgical therapy is critical. However, mortality may be significant, especially in patients with neutropenia. Leukocyte response to the infection may be a prognostic marker. Pseudomonas and enteric gram-negative organisms are seen frequently in immunocompromised children with necrotizing fasciitis. Antimicrobial selection should supply adequate coverage of these organisms. PMID- 7472966 TI - Tobogganing injuries in children. AB - AIM: To undertake a retrospective review of tobogganing-related injuries in children requiring admission to the Hospital for Sick Children, Toronto. MATERIALS AND METHODS: Twenty-two children with tobogganing-related injuries were admitted between December 1991 and December 1993. There were 13 boys and 9 girls (median age, 10 years; range, 3 to 17 years). Their charts were reviewed using a structured proforma. RESULTS: Nine patients (41%) sustained injuries by striking a tree, 8 struck other objects, and 5 fell from the toboggan. Only 1 was wearing protective headgear. The initial site of impact was the head in 13 patients (59%), the trunk in 5, and the extremities in 4. Major injuries were sustained in all body systems. Thirteen patients (59%) required surgical treatment and two patients died, one of cerebral edema and the other of acute renal failure and subsequent multiorgan failure. CONCLUSION: Tobogganing-related injuries represent a small fraction of all injuries in children who require hospitalization. However, such injuries can be serious and can cause significant morbidity and mortality. Better public awareness of the risks of tobogganing is required and simple safety guidelines should be developed to reduce the risks of this popular winter pastime. PMID- 7472967 TI - An intraluminal model of necrotizing enterocolitis in the developing neonatal piglet. AB - The most common risk factors for the development of necrotizing enterocolitis (NEC) are prematurity and enteral feeding. Most models of NEC involve ischemic insult resulting in generalized necrosis, different from the classical ileocecal predilection of NEC. This anatomic predisposition is explained by dysmotility of immature gut, leading to bacterial overgrowth in the terminal ileum and colon. Infant formula containing lactose as the sole carbohydrate source overwhelms partially developed lactase activity, allowing enteric bacteria to ferment excess carbohydrate to short-chain fatty acids, decreasing intraluminal gut pH and predisposing to mucosal injury. Impaired clearance of intraluminal contents exacerbates this effect. In the present study the authors used a model of NEC, originally developed in rabbits and based on analysis of intestinal contents of NEC babies, modified and adapted here to neonatal piglets, the gastrointestinal tract of which more closely resembles the human neonate. METHOD: Piglets < 3 days old and 2 weeks old were laparotomized. Loops created from the distal ileum to the proximal colon were injected with isoosmolar acidified casein solution or 0.9% saline. Segments were harvested 3 hours later, sectioned for H&E, and graded from 0 (intact villi) to 4 (transmural necrosis). RESULTS: Acidified casein induced damage included areas of necrosis, submucosal edema, inflammatory cell infiltrate, and lymphatic distension. In younger animals, lesions were more pronounced (3.25 +/- 0.13 for the < 3-day-old v 2.43 +/- 0.14 for the 2-week-old piglets; P < .005). CONCLUSION: The authors believe that this piglet NEC model most closely approximates human NEC because it incorporates two of the most common risk factors: dysmotility (by creating intestinal loops) and enteral feeding (by intraluminal injection of acidified casein). PMID- 7472968 TI - The principles of normal and abnormal hindgut development. AB - In the past, several theories have been proposed to explain the occurrence of anorectal malformations. Most investigators believe that these malformations are the result of an impaired process of septation. However, in 1986 vd Putte challenged all theories that tried to explain anorectal malformations by a faulty fusion of lateral ridges of the cloaca. To elucidate the principles of normal and abnormal cloacal development, the authors studied the morphology of this region in normal embryos of rats and abnormal embryos of SD mice, which often have abnormal cloacas. Using scanning electron microscopy (SEM), 245 normal rat embryos and 80 abnormal SD-mice embryos were observed. The results were as follows. (1) In normal embryos the region of the future anal opening can be identified soon after the establishment of the cloacal membrane. This part is a fixed point in cloacal development. (2) In abnormal embryos the cloacal membrane is too short. The region of the future anal opening is missing. (3) In abnormal embryos a spectrum of malformed cloacas can be observed. This is in accordance with the spectrum of anorectal malformations clinically observed in humans. (4) The authors' observations support recent findings that the "fistula" in anorectal malformations resembles a normal anus at an ectopic position. PMID- 7472969 TI - Button-pexy fixation for repair of ileostomy and colostomy prolapse. AB - Intestinal stomas are used in the surgical management of a number of congenital and acquired gastrointestinal conditions. Minor stomal complications are relatively common and are often well managed with nonoperative measures. Stomal prolapse may not represent a serious problem, but occasionally it requires surgical correction because of severe skin excoriation, bleeding, or incarceration of the bowel. The authors describe the treatment of stomal prolapse with the technique of button-pexy fixation, as originally described by C.W. Mayo. PMID- 7472970 TI - The incidence and spectrum of neurological injury after open fetal surgery. AB - A preterm infant's immature brain is susceptible to both anoxic and hemorrhagic injury during periods of physiological stress. The advent of in utero surgery has created a new population of premature patients at risk for central nervous system (CNS) injury. The aim of this study was to evaluate the frequency and nature of CNS injuries in fetal surgical patients. Of 33 fetuses with known neurological outcome after fetal surgery, CNS injuries were identified in seven (21%). Of the seven, four had significant episodes of fetal bradycardia (3) or neonatal hypotension (1), which suggests that asphyxia contributed to the neurological injury. The CNS injuries in the other three patients occurred unexpectedly and without associated signs of fetal distress. The authors speculate that these injuries may have been caused by sudden fluxes in cerebral blood flow, induced by maternal hypoxia (1) or by maternally administered tocolytic drugs (2) used to treat postoperative preterm labor. PMID- 7472972 TI - Sclerosing of recurrent lymphangioma using OK-432. AB - The authors present two cases of lymphangioma of the cervicofacial region, treated with a new investigational drug in North America, OK-432 (picibanil), a sclerosing agent intralesionally injected. Both patients had been treated surgically and had recurrence of the tumor. Intralesional injection of OK-432 without aspiration was employed for the first patient, and after aspiration in the second patient. A change in consistency of the tumor, manifested by softening, was followed by marked shrinkage. No complication either locally or systemically has occurred during the follow-up period (10 to 16 months). In both cases, satisfactory results were obtained, resulting in definite reduction in size and improvement in cosmetic appearance. The authors recommend OK-432 intralesional injection for surgically challenging lymphangioma. Their results support those of a recent Japanese study using OK-432 as sclerosing therapy for unresectable lymphangioma. PMID- 7472971 TI - Effects of intraamniotic helium, carbon dioxide, and water on fetal lambs. AB - The hysteroscopic approach to the correction of congenital anomalies in utero may be helped by the temporary introduction of clear, electrically neutral fluids (liquids or gases) into the amniotic cavity. Accordingly, the authors determined the effects of three candidate fluids (helium, carbon dioxide, and water) on the oxygenation, acid-base balance, electrolyte concentrations, and hemodynamic function of nine sheep fetuses at 120 days' gestation (full term 145 days). Each fetus was exposed, in random order, to each of the three fluids, at an intraamniotic pressure of 15 mm Hg. The authors found that helium produced no changes in any of the measurements and was well tolerated by the fetus and the mother. Carbon dioxide decreased fetal arterial pH, and increased both arterial PCO2 and PO2. Although the decrease in pH was explained entirely by the increase in PCO2, the increase in arterial PO2 may have resulted from vasodilatation of the placental circulation or vasoconstriction of the fetal circulation with a redistribution of blood flow to the placenta. Water caused mild fetal hyponatremia and hypochloremia, without significant changes in acid-base balance or oxygenation. The results suggest that both helium and water may represent viable alternatives for amniotic fluid replacement during hysteroscopic surgery. PMID- 7472973 TI - Stratification of injury severity using energy expenditure response in surgical infants. AB - AIM OF STUDY: Injury severity stratification has important clinical outcome significance and can influence nutritional management. Although surgery alone has been shown not to increase measured energy expenditure (MEE) substantially, large increases in MEE can result from severe underlying acute illness, which frequently necessitates surgery (like sepsis or intense inflammation). The authors hypothesized that the magnitude and duration of the MEE response to surgery associated with a severe preoperative acute injury would exceed that of surgery in which no substantial preoperative stress was present, thus representing an index of overall injury severity in surgical infants. METHODS: MEE (kcal/kg/d) was determined on postoperative days (POD) 2, 5, and 8 in 12 infants (average age, 47 days) after two separate injury insults (at least 8 days apart). In each patient, one operation resulted in a peak serum C-reactive protein (CRP) concentration of less than 6.5 mg/dL (low stress), and the second operation, preoperatively associated with sepsis or a major inflammatory insult, resulted in a peak CRP of more than 6.5 mg/dL (high stress). Data were paired so that each child served as his or her own control. The initial basal protein calorie delivery was similar in both groups. MAIN RESULTS: The mean peak CRP values were 14.1 +/- 10.7 mg/dL (high stress) and 4.1 +/- 2.3 mg/dL (low stress) and returned to normal levels earlier (before POD 8) after injury insult in the low-stress group. Analysis of energy expenditure on POD 2 demonstrated significantly elevated mean MEE values in the high-stress group (58.0 +/- 12.2 kcal/kg/d v 39.4 +/- 9.5 kcal/kg/d in the low-stress group; P = .0001). In contrast, analysis of POD 8 energy expenditure showed significantly lower mean MEE values in the high-stress group (50.7 +/- 12.0 kcal/kg/d) v (66.4 +/- 15.1 kcal/kg/d in the low-stress group; P = .0118) group. CONCLUSION: The early (POD 2) hypermetabolic response to injury as determined by MEE effectively differentiated the two stress groups. This finding suggests that acute underlying illness is an important determinant of postoperative MEE. Furthermore, in the low stress group, serial CRP levels returned to normal earlier, associated with significantly greater late (POD 8) MEE values. Because MEE is directly proportional to growth rate in healthy infants, and growth is retarded during acute metabolic stress, these findings suggest that increased energy is utilized for growth recovery following the earlier resolution of the acute injury response in the low-stress group. These data indicate that serial postoperative MEE can be used to stratify injury severity and may be an effective parameter to monitor the return of normal growth metabolism in surgical infants. PMID- 7472974 TI - Fetal endoscopic ('Fetendo') surgery: the relationship between insufflating pressure and the fetoplacental circulation. AB - Application of video-endoscopic surgery to the gravid uterus provides a new treatment option for the fetus with a correctable congenital anomaly. "Fetendo" surgery requires temporary enlargement of the uterine cavity to create a working space. Volume expansion of the amniotic space raises intrauterine pressure, which could increase placental vascular resistance and thereby reduce placental blood flow. To test this hypothesis, the authors developed a fetal sheep model to examine the relationship between insufflating pressure and flow in the placental circulation. Fetoplacental blood flow was measured via ultrasonic flow probes placed around the fetal common umbilical artery and the maternal uterine artery in five anesthetized 120-day-gestation ewes. Invasive feto-maternal monitoring permitted synchronous measurement of fetal mean arterial pressure, fetal central venous pressure, maternal mean arterial pressure, amniotic pressure, and fetal oxygen saturation, with calculated values for fetal and maternal placental vascular resistance. Amniotic pressure was raised from 10 mm Hg to 40 mm Hg in 5 mm Hg increments by a combination of saline amnioinfusion and external uterine compression. At amniotic pressures of 20 mm Hg or less, placental blood flow was preserved; however, elevation of amniotic pressure above 20 mm Hg resulted in a significant decrease in placental flow, with concomitant fetal hypoxia. The authors conclude that the relationship between intrauterine pressure, flow in the placental circulation, and fetal oxygen delivery must be considered when selecting intrauterine insufflation pressures for hysteroscopic intervention. PMID- 7472975 TI - Gastroschisis: a simple technique for staged silo closure. AB - In conjunction with the Neonatology Department at Loma Linda University Children's Hospital, a new protocol has evolved for the management of infants with gastroschisis, which obviates both risks associated with primary and staged silo closure. After stabilization of the infant in the neonatal intensive care unit, under sterile conditions, a 5- or 7-cm SILASTIC silo with a spring-loaded ring is placed over the exposed viscera, under the fascial defect. No sutures are required. A fentanyl drip is given, and the bowel is gradually reduced over the next few days. The transparent material of the silo allows for continuous monitoring of the condition of the bowel. Second-stage closure in the operating room is performed using a purse-string suture in the fascia to create a pseudoumbilicus. From October 1992 to April 1994 the authors managed 10 infants using this protocol. The results are compared with those of infants with gastroschisis treated at the same institution between August 1982 and June 1993. Outcome parameters to be compared include time until closure, time on ventilation, days of total parenteral nutrition, time until start of oral feeding, time until toleration of full-volume oral feeding, and time until discharge. The authors conclude that silo closure in the neonatal intensive care unit is simple, quick, and effective. It eliminates multiple trips to the operating room, allows the natural accommodation of the bowel into the abdominal cavity with little edema and minimal vascular compromise, and has become the authors' treatment of choice for infants with gastroschisis. PMID- 7472976 TI - Reversible tracheal obstruction in the fetal sheep: effects on tracheal fluid pressure and lung growth. AB - Congenital diaphragmatic hernia (CDH) continues to carry high morbidity and mortality. A number of treatment modalities including extracorporeal membrane oxygenation and in utero repair have improved the mortality rate only minimally. With this condition, there is often insufficient lung mass at birth and persistent pulmonary hypertension postnatally. Experimental fetal tracheal ligation has been shown to increase lung growth in utero and to reduce the hernial contents in CDH. The purpose of this study was to determine the effect of reversible fetal tracheal occlusion on tracheal pressure and lung development. Nine fetal sheep were divided into two groups. Group 1 had intratracheal balloons placed, and the balloons were left inflated for 21 to 28 days. Group 2 consisted of littermates that served as controls. They either had uninflated balloons placed or were left unoperated. Tracheal pressure measurements were recorded periodically, and the amniotic fluid pressure served as a reference. The animals were killed near term, and the lungs, heart, and liver were weighed and corrected for body weight. Standard morphometry was used to compare the lungs further, and the lung DNA and protein content were measured. Tracheal damage from the balloon catheter also was assessed. The tracheal pressure was 3.85 (+/- .49 SE) mm Hg in experimental animals, and it averaged -0.27 (+/- .27 SE) mm Hg in controls (P < .0001). Tracheal occlusion increased lung weight and volume by two to three times (P < .0001 and P < .0006, respectively) while heart and liver weights remained similar to those of controls.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7472977 TI - Combined gas ventilation and perfluorochemical tracheal instillation as an alternative treatment for lethal congenital diaphragmatic hernia in lambs. AB - Tracheal instillation of perfluorochemical liquid (PFC) lowers surface tension in the lung and thus might reduce barotrauma commonly associated with conventional gas ventilation (GV) in highly immature and hypoplastic lungs. It could be a promising alternative treatment for congenital diaphragmatic hernia (CDH) when GV alone is proving inefficient. The authors compared data for eight newborn lambs with surgically induced CDH. The animals had GV and were studied (in 2 groups) for up to 3.5 hours. Group 1 (GV, n = 4) had gas ventilation only. In group 2 (PFC, n = 4), after 30 minutes of GV, 10 to 12 mL/kg of warmed, oxygenated PFC liquid (LiquiVent) was instilled into the lung via the trachea under pressure volume curve monitoring. Arterial pressure, blood chemistry, and pulmonary mechanics were evaluated serially; histological analysis was performed. One preassigned animal in group 1 died after 15 minutes. After 30 minutes of life, the cardiopulmonary profile of survivors was indicative of severe respiratory distress (Pao2 < 72 mm Hg with FIO2 at 1.0, PaCO2 > 90 mm Hg, compliance < 0.10 mL/cm H2O/kg) and not different between groups; the severity of pulmonary hypoplasia was further confirmed postmortem; the ratio of lung weight to body weight was 41% of that observed in control lambs, in both gas-only and combined gas/PFC-ventilated animals, compared with their respective controls. After instillation of PFC, there were dramatic improvements in acid-base status and pulmonary compliance in group 2. Survival at 3.5 hours also was markedly different (4 of 4 PFC animals and 1 of 3 GV animals).(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7472978 TI - Timing of repair of congenital diaphragmatic hernia requiring extracorporeal membrane oxygenation support. AB - Treatment of congenital diaphragmatic hernia (CDH) has undergone a revolutionary change in philosophy, from previous urgent repair to the present practice of stabilization and delayed repair. However, when extracorporeal membrane oxygenation (ECMO) is required, many people believe that the risk of postoperative pulmonary hypertension (PPHN) mandates hernia repair while on ECMO. This report details the experience in two ECMO centers with stabilization, ECMO if required, and CDH repair post-ECMO. All CDH patients symptomatic in the first hour of life with a gestational age of at least 34 weeks during the period were reviewed retrospectively. Standard criteria were used to select patients for ECMO. High-frequency jet or oscillating ventilators and nitric oxide were not routinely available throughout the study period, but were used in some of the more recent patients. A total of 60 patients presented to the two centers; 24 cases were stabilized with conventional management, repair of the CDH was done elective, and survival was 100%. Eight patients were referred after having repair elsewhere; six survived (75%). The two deaths were attributable to associated lethal lesions--complex cyanotic heart disease and alveolar capillary dysplasia. Eight patients who required ECMO were managed with the intention of repairing the defect on ECMO. Four survived (50%). Two patients died before repair. Twenty patients were managed with ECMO, with the intention of repairing the defect after decannulation. Overall survival was 13 (65%), deaths were caused by pre-ECMO hypoxia, pulmonary insufficiency, and associated cardiac disease. No patient had recurrent pulmonary hypertension after late repair.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7472979 TI - Survival in neonatal congenital hernia without extracorporeal membrane oxygenation support. AB - The experience with high-risk congenital diaphragmatic hernia (CDH) at an institution that does not offer extracorporeal membrane oxygenation (ECMO) was reviewed. Between January 1, 1983 and December 31, 1993, 38 children presented with Bochdalek-type CDH. Excluded were two infants with lethal cardiac anomalies and four presenting after 4 hours of age. Thus, the authors identified 32 high risk patients. All had early respiratory distress and were intubated within 5 hours of birth. Sixteen were inborn; 16 were referred to the Izaak Walton Killam Children's Hospital (IWK) within 24 hours of birth. There were 19 males and 13 females. Three died before surgery could be attempted. Twenty-two survived, giving an overall survival rate of 69% (22 of 32). For 28 of the 32, the best preoperative postductal Pao2 (BPDPao2) was recorded. Fifteen of the 28 children had a BPDPao2 of greater than 100 mm Hg. Survival in this group was 14 of 15 (93%). Thirteen of the 28 patients had a BPDPao2 of less than 100 mm Hg. Survival in this group was 5 of 13 (38%). These survival rates are comparable to those of centers offering ECMO. BPDPao2 appears to be a useful discriminating variable. PMID- 7472981 TI - Testicular fate after incarcerated hernia repair and/or orchiopexy performed in patients under 6 months of age. AB - This study assessed testicular viability after 476 inguinal procedures performed in 338 infants under 6 months of age, between 1974 and 1993. One hundred twenty one elective hernia repairs (contralateral explorations) were compared with 355 hernia repairs complicated by incarceration and/or orchiopexy. Clinical examination of 323 testes in the early postoperative period showed 20 atrophic testes. Since January 1994, 71 of the 338 patients have had testicular measurements obtained through ultrasonography (US). An additional 13 atrophic testes were found during US examination. Of these, nine were believed to be normal during early postoperative examination. Assuming that US examination will confirm atrophy in the 20 atrophic testes noted early in the postoperative clinical evaluation, and that all other testes not yet scanned are found to be normal, the minimal atrophy rate (MAR) would be 9.3% (33 of 355). Neither operative nor early postoperative testicular assessment correlates with ultimate testicular survival. Testicular pathology may become more evident after puberty, and the real incidence of atrophy may increase. PMID- 7472980 TI - Pathophysiology of congenital diaphragmatic hernia. XVI: Elevated pulmonary collagen in the lamb model of congenital diaphragmatic hernia. AB - The pathophysiologic features of congenital diaphragmatic hernia (CDH) include pulmonary hypoplasia, pulmonary hypertension, surfactant deficiency, and decreased pulmonary compliance. When the surfactant deficiency is corrected using exogenous surfactant therapy, the pulmonary compliance improves, but does not reach normal values. Quasistatic saline pressure-volume measurements, which eliminate the air-liquid interface, confirm that CDH lungs are intrinsically less compliant than control lungs. The authors hypothesized that this abnormal lung compliance results from elevated concentrations of collagen and/or elastin in the lung. Therefore, they measured the collagen and elastin concentrations in CDH and control lung tissue. Also measured was the collagen concentration in the kidney, intestine, and dissected third-generation arterioles, venules, and bronchioles, to characterize further the pathology of CDH. The CDH model was created on the left side of fetuses in pregnant ewes at 80 days' gestation. The fetuses were delivered and killed at 140 days (full term, 145). The concentrations of collagen (as hydroxyproline), elastin, DNA, and total protein were measured using standard techniques. Although there was significantly more collagen per gram of lung tissue in the CDH lungs (1.334 mg/g v 0.885 mg/g in the controls) the elastin concentrations were not different. The elevated collagen concentration was not associated specifically with the conducting airways or vasculature. The collagen concentrations in CDH kidneys and intestines were the same as those of controls. The DNA/total protein ratios in the CDH and control lungs were identical. The results suggest that the elevated collagen concentration was present only in the lungs of CDH lambs, and that it was not attributable to atrophy or hypertrophy of the lungs. Thus, increased collagen in the lung parenchyma may be responsible for the intrinsic stiffness and decreased compliance of the CDH lungs. PMID- 7472982 TI - Long-term venous access using endogenous splenic tissue: the 'spleen-o-port'. AB - The authors sought to determine whether endogenous splenic tissue placed in a subcutaneous pouch ("spleen-o-port") could function as a viable alternative to central venous catheters/ports for long-term venous access. A small transverse incision was made in the left upper quadrant of each puppy (n = 6) under general anesthesia. Using a stapler, the authors divided the splenic parenchyma. The superior portion was returned to its native location, and a subcutaneous pocket was created to house the inferior pole with its attached vascular supply. The fascial and muscular layers were closed with care to avoid compressing the blood supply to the spleen-o-port. Postoperatively the dogs resumed normal activity. There have been no deaths, infectious complications, splenic ruptures, or thromboses over a 6-month period. Under fluoroscopy, the dogs were imaged from postoperative day (POD) 10 to 177. Contrast agent entering the splenic parenchyma was promptly visualized in the splenic vein and then filled the portal vein. Electrolyte measurements from spleen-o-port blood samples were identical to those from peripheral venous samples. After gentamicin (mixed in a crystalloid solution) was infused through the spleen-o-port, the peak serum level corresponded to the therapeutic levels observed after standard intravenous administration. The spleen-o-port permits rapid infusion of drugs and crystalloid, and allows repetitive blood sampling while eliminating the foreign body that can promote septicemia in the immunocompromised patient. PMID- 7472983 TI - Pediatric laparoscopic splenectomy. AB - Laparoscopic surgery is becoming widely accepted as an alternative to conventional procedures. It is becoming more and more evident that laparoscopic techniques can be applied successfully to pediatric patients. Advantages of these techniques include less postoperative pain, decreased ileus, fewer pulmonary complications, and shorter hospital stays. Elective splenectomy for hematologic disease or for staging of Hodgkin's lymphoma also appears to be amenable to laparoscopic techniques. This report details 12 consecutive splenectomies successfully performed laparoscopically since July 1993. No case required conversion to laparotomy. Each case was reviewed with respect to operative time, estimated blood loss, identification of accessory spleens, time until full oral intake, analgesia requirements, and length of stay. Factors contributing to morbidity such as ileus, pulmonary complications, and would infections were evaluated. Documentation was also reviewed for late sequelae such as intestinal obstruction and incisional hernias. These patients were compared with 20 consecutively treated patients who underwent open splenectomy in the period immediately preceding the use of laparoscopic splenectomy. Laparoscopic splenectomy, in the authors' experience, is a safe alternative to open splenectomy, has few complications, is cost effective, and has been well accepted by patients and families. PMID- 7472984 TI - Gasless laparoscopy in infants: the rabbit model. AB - Laparoscopic operations can be performed in neonates and infants, but carbon dioxide pneumoperitoneum may be more dangerous than in adults. Therefore the concept of gasless laparoscopy is especially attractive for small children. The authors have developed an animal training model of gasless infant laparoscopy using the rabbit. Eleven New Zealand white rabbits (mean weight, 2.2 kg) were premedicated with fentanyl (10 micrograms intramuscularly) and maintained under inhalation anesthesia by mask, with halothane (1.0 to 2.5%) in oxygen and nitrous oxide (1:1). The animals were neither intubated nor mechanically ventilated. Heart rate and oxygen saturation were monitored by pulse oximetry. A supraumbilical incision was made through the peritoneum, and an abdominal wall elevator was inserted. A 4-mm-diameter 30 degrees endoscope was introduced at the same site, allowing excellent visualization of the abdominal cavity. Three additional 5-mm cannulas were placed, and a standard Nissen fundoplication was performed in all 11 animals. In seven of these, a left nephrectomy was also performed. Five-millimeter pediatric instruments and 1.7- and 2.7-mm laryngoscopic instruments were used. The rabbit can serve as a training model for laparoscopy in the infant. Gasless laparoscopy, using abdominal wall elevation, may be particularly useful in very small patients. PMID- 7472985 TI - Laparoscopic-assisted percutaneous endoscopic gastrostomy. AB - The authors report the successful use of laparoscopic-assisted percutaneous endoscopic gastrostomy (LAPEG) in two children. Attempts at simple percutaneous endoscopic gastrostomy in both patients had failed. Subsequently, LA-PEG was easily accomplished. This technique consisted of a combination of upper gastrointestinal endoscopy and laparoscopy. The gastrostomy was placed under direct vision in the lesser gastric curvature, hence minimizing the risk of developing gastroesophageal reflux. PMID- 7472986 TI - Thoracoscopy in the management of pediatric empyema. AB - The surgical management of empyema consists of (1) aggressive therapy with thoracotomy and decortication or (2) conservative treatment with chest tube drainage and intravenous antibiotics. Recently, Kern and Rodgers introduced thoracoscopic debridement as an adjunct to the management of children with empyema, with promising results. Hence, the authors report their experience with thoracoscopy in the management of pediatric patients with empyema. In the last years, 10 children have undergone thoracoscopic debridement (TD) for empyema. The average age was 6.9 years (range, 2 to 16). Children underwent TD an average of 14 days (range, 8 to 16) after initial presentation and 4 days (range, 2 to 6) after admission to the authors' hospital. Indications for TD were persistent requirement of supplemental oxygen and failure of conservative medical management that consisted of antibiotics and tube thoracostomy. Three children had positive pleural fluid cultures for Streptococcus pneumoniae. In all cases, preoperative ultrasound or chest computed tomography examination showed dense pleural fluid with septation. During surgery, TD allowed for lung expansion and precise chest tube placement in all patients except one who required conversion to minithoracotomy and decortication for persistent encasement with a thick pleural peel. There were no postoperative complications related to the procedure. After TD, all children had prompt clinical improvement. The patients were weaned from supplemental oxygen by postoperative day 2, and following early chest tube removal, nine children were discharged home by postoperative day 7 (range, 3 to 10). One child required further hospitalization for underlying renal failure. In the authors' hands, TD was effective in producing prompt clinical improvement in children with empyema.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7472987 TI - Colonic atresia combined with Hirschsprung's disease: a diagnostic and therapeutic challenge. AB - Three new cases of patients with co-occurrence of colonic atresia and Hirschsprung's disease and a review of the literature (n = 5) are presented in this report. All patients (n = 8) except one were full-term infants who had no other significant anomalies. The preterm infant had associated tetralogy of Fallot in addition to Hirschsprung's disease and colonic atresia. Six patients had atresia of the ascending colon, and two had atresia of the colon to splenic flexure. All colonic atresias were diagnosed neonatally; however, there was mean delay of 15.4 months (range, 1 mo to 5 years) in diagnosing associated Hirschsprung's disease. There were two deaths. A careful examination of the resected specimen to rule out Hirschsprung's disease is recommended. Performing a rectal biopsy must be considered for patients who initially were treated for colonic atresia and who have a slow return of normal gut function. PMID- 7472988 TI - The complete spectrum of neurocristopathy in an infant with congenital hypoventilation, Hirschsprung's disease, and neuroblastoma. AB - Neuroblastoma, Hirschsprung's disease, and central hypoventilation (Ondine's curse) are considered aberrations of neural crest cell growth, migration, or differentiation, and as such are considered to be under the general heading of neurocristopathy. Their combined occurrence in a newborn infant presenting with total colonic aganglionosis, central hypoventilation, and multifocal neuroblastoma had not been reported previously. A 2.3-kg white full-term girl required endotracheal intubation because of persistent apnea in the first hours of life. She had progressive abdominal distension and failure to pass meconium; a barium enema was performed, which showed microcolon with meconium pellets at the distal ileum. During laparotomy the distal ileum was found to be obstructed with inspissated meconium; an ileostomy and appendectomy were performed. The resected specimens were aganglionic. An additional 20 cm of aganglionic ileum was removed, and a normally innervated ileostomy was constructed. Numerous attempts at extubation failed because of apnea. The results of an extensive apnea workup, including electroencephalogram, magnetic resonance imaging (MRI), bronchoscopy, and pH probe study, were normal. Sleep studies showed congenital central hypoventilation syndrome, and the patient underwent a tracheostomy. At 3 months, an abdominal ultrasound examination performed within a septic workup showed a right suprarenal mass extending across the midline. Thoracic and abdominal MRI scans showed large bilateral adrenal and posterior mediastinal masses. The serum catecholamines and ferritin level were markedly elevated, suggestive of neuroblastoma. In light of the child's multiple problems, the family chose to forgo further workup (including a tissue biopsy) and therapy. In the following 2 months her tumor load rapidly progressed, and she died of respiratory insufficiency.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7472989 TI - Long-term complications in cases of isolated esophageal atresia treated with esophageal anastomosis. AB - Long-term follow-up examination with esophagogastroduodenoscopy was performed on seven patients who had undergone successful delayed anastomosis for isolated esophageal atresia. The follow-up period ranged from 1.2 to 11.3 years (mean, 5.3). All patients had undergone fundoplication because of symptomatic gastroesophageal reflux (GER). Three anastomotic strictures had to be resected. At the time of the last follow-up examination, the subjective results were excellent for five patients and good for two. The last endoscopy showed macroscopic esophagitis in three and normal mucosa in four. The fundoplication was partly disrupted in two patients. In three patients the fundoplication was competent but partly intrathoracic. Histological examination showed moderate esophagitis in one, mild esophagitis in one, and normal mucosa in five patients; however, four patients were on continuous medication for esophagitis. In conclusion, the subjective results of patients with isolated esophageal atresia treated with esophageal anastomosis are good. However, long-term complications caused by GER are common in these patients. Therefore, active search and treatment of reflux is necessary for these patients. PMID- 7472990 TI - Ambulatory 24-hour manometric and pH metric evidence of permanent impairment of clearance capacity in patients with esophageal atresia. AB - Dysphagia and gastroesophageal reflux (GER) probably caused by structural disorganization of the esophagus occur frequently after repair of tracheoesophageal fistula (TEF), and the extent to which they may improve beyond childhood is not known. The aim of the present study is to assess by combined ambulatory 24-hour manometry and pH-metry the esophageal peristaltic activity and acid clearing capacity in adolescents and adults who had been operated on for TEF at birth. Twenty-two patients, aged 17.1 +/- 4.5 years (mean +/- SD), were examined with combined three-channel manometry and two-channel pH-metry followed by endoscopy and biopsy. Although they considered themselves healthy, on careful interrogation, 16 (72%) were found to have dysphagia, 13 (59%) had heartburn, 10 (45%) had foreign body impaction, and 7 (31%) had chronic respiratory tract disease. GER was detected in 12 (54%) patients (5 with histological esophagitis), 10 of whom had a pattern of prolonged nocturnal episodes with very slow clearance. All patients had diminished contractile activity with low-amplitude and short-duration waves that decreased from 0.53 +/- 0.35 waves per minute to 0.28 +/- 0.2 waves per minute during sleep. Propulsive activity was uniformly disorganized, with peristaltic sequences being few (less than 50% overall) and incomplete (above 80%). Finally, the acid-clearing capacity was nil; the proportions of ineffective sequences were above 90% for all periods considered, including sleep and mealtimes. The motor behavior of nonrefluxing and refluxing patients was identical despite the differences in esophageal acid exposure.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7472991 TI - Application of Collis gastroplasty to the management of esophageal atresia. AB - Establishment of esophageal continuity in the presence of an isolated esophageal atresia frequently requires deferred esophageal substitution. The author describes the use of Collis gastroplasty as an esophageal lengthening technique for the distal esophagus. This technique permitted the performance of primary repair of the esophagus in an infant with a 6-cm gap that otherwise would have required an esophageal substitution procedure. Morbidity after this procedure is comparable to that occurring after a number of established techniques. PMID- 7472992 TI - Endoscopic closure of recurrent tracheoesophageal fistula using Tisseel. AB - The author reports on two patients who presented with recurrent tracheoesophageal fistula. They were treated with the endoscopic application of fibrin sealant (Tisseel) directly to the fistula tract. The technique may require multiple applications before complete fistula closure is achieved. The procedure is associated with low morbidity and may eliminate the need for a second thoracotomy. PMID- 7472993 TI - Isolated congenital esophageal stenosis. AB - The incidence of congenital esophageal stenosis (CES) is approximately 1 in 25,000 to 50,000 live births. There is associated esophageal atresia in one third of cases; the remainder are classified as isolated CES. Histologically, the anomaly may include tracheobronchial remnants, a membranous diaphragm, or diffuse fibrosis of the muscularis and submucosa. The authors report their experience with three patients who had isolated CES. The patients were free of symptoms for the first 6 months of life. All had difficulty with feeding from 6 to 12 months of age, which corresponded with the introduction of solids. Most solid feedings were regurgitated. Evaluation consisted of cine-esophagogram, pH monitoring, manometry, and endoscopy, with biopsies to exclude the diagnosis of gastroesophageal reflux. All patients underwent hydrostatic dilatation, but the benefit was only transient. The patients were referred for surgical correction when symptoms recurred. Limited resection of the esophageal stenosis with primary anastomosis was performed on all three patients via a left thoracotomy. The stenoses were located in the distal third of esophagus, near the junction with the middle third. Pathological examination showed tracheobronchial remnants in one patient and fibrotic muscle in the other two. The average age at the time of surgery was 19 months. The hospital stay averaged 8 days. The contrast study 1 week postthoracotomy showed esophageal patency in all patients, with no leakage. All were discharged from the hospital, tolerating solid food. CES should be sought for in patients who present with dysphagia to solid food that begins after the first 6 months of life.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7472994 TI - Colon patch esophagoplasty for caustic esophageal stricture. AB - Severe caustic injury with stricture may require esophageal reconstruction. The available methods of esophageal substitution do not satisfactorily replace the peristaltic and antireflux properties of the native esophagus. The authors report the results of two children treated with colon patch esophagoplasty and preservation of the injured esophagus. Both were 2 years of age when they accidently swallowed lye. Long esophageal strictures developed, which were resistant to repeated dilatation. Eleven and 13 months after the injury, the patients underwent colon patch esophagoplasty. A segment of the left colon was tailored to the length and shape of the opened stricture and was anastomosed side to-side to the esophagotomy. One child later had a short recurrent stricture and diverticulum, which required revision of the distal end of the anastomosis. Follow-up endoscopic examination has shown healing of the esophagus and resolution of the strictures; both children are eating normally 3 years after surgery, without symptoms of gastroesophageal reflux. Colon patch esophagoplasty is an alternative to esophageal replacement for long caustic esophageal strictures. Its advantage is preservation of the peristaltic and antireflux mechanisms of the conserved esophagus. PMID- 7472995 TI - The safety of home tracheostomy care for children. AB - Home tracheostomy care avoids prolonged hospitalization for tracheostomized children who must remain cannulated for a long time; however the safety aspects of this form of management are still under discussion. Since 1987 the authors have tutored parents in tracheostomy care. Once parental training was complete and the necessary equipment (aspirator, humidifier, etc) provided, children in stable condition were discharged and had period follow-up as outpatients. Thirty four children had a total of 710 months in home care; 11 of them had decannulation. The only reported complications were two partial obstructions of the cannula and two accidental decannulations. The low number of documented accidents and the parents' profound appreciation in having their children at home indicate that patients with a tracheostomy in need of long-term care can be safely managed at home. PMID- 7472996 TI - Effects of FMLP and LPS on [Ca2+]i of peritoneal exudate polymorphonuclear leukocytes following onset of inflammation. AB - Because a general study of activated neutrophils may have relevance to periodontal diseases and accompanying inflammation, we studied a function of mouse polymorphonuclear leukocytes (PMNs) that exude into the peritoneal cavity in response to inflammation caused by i.p. injection of 2% casein. The effects of E. coli-lipopolysaccharide (E-LPS) and a chemotactic factor, N-formyl-N-methionyl N-leucyl-L-phenylalanine (FMLP), on the level of intracellular calcium ([Ca2+]i) in these PMNs were examined. From analysis made with a laser cytometer (ACAS 570), the PMNs in exudates harvested 3-9 h after the onset of inflammation were shown to undergo [Ca2+]i elevation in response to 10(-6) M FMLP. The peak concentration of [Ca2+]i elicited by FMLP was highest in exudate cells 6 h after casein injection. In addition, about 65% of the PMNs in the 3-h exudate were FMLP sensitive displaying an elevated [Ca2+]i, whereas more than 85% of them in 6- and 9-h exudates became FMLP sensitive. Also, the maximum level of [Ca2+]i after FMLP stimulation was potentiated by pretreatment of the cells with E-LPS (0.2 microgram/ml). The present study suggests that PMNs induced by casein injection and appearing in mouse peritoneal exudate at different times possess significantly different ability to undergo [Ca2+]i elevation, and different susceptibility toward a chemotactic factor, FMLP. PMID- 7472997 TI - Presence of activated eosinophils, high IgE and sCD23 titers in gingival crevicular fluid of patients with adult periodontitis. AB - Our previous studies showed that the expression of CD23 on polymorphonuclear leukocytes (PMNLs) in gingival crevicular fluid (GCF) from adult periodontitis (AP) patients was higher than in autologous peripheral blood (PB). Percentages of eosinophils in GCF PMNLs ranged between 6 and 14%. The purpose of the present studies was to increase understanding of the potential role of eosinophils and their products, including CD23, in periodontal disease. We analysed the eosinophil fraction in GCF and PB by flow cytometry using monoclonal antibodies to CD23b (BB10), eosinophil cationic protein (ECP) in stored and secretory forms (EG1 and EG2), and CD67 (80H3). Simultaneously, we measured IgE and soluble CD23 titer and GCF and serum by ELISA. Flow cytometric analysis of BB10, EG2 and 80H3 binding showed that GCF eosinophils from AP were activated. A large BB10+ EG2+ cellular fraction was detected in GCF from AP whereas it was very low in autologous serum (9.30 +/- 2.460 vs 0.16 +/- 0.10, p < 0.001). GCF from gingivitis patients exhibited no flow cytometric evidence for the presence of BB10+ EG2+ cells. BB10+ EG1+ cells, or inactivated eosinophils rated lower in GCF than in PB both in gingivitis and periodontitis patients (0.45 +/- 0.63 vs 1.83 +/- 0.96 and 0.15 +/- 0.30 vs 1.30 +/- 0.20, p < 0.05, respectively). IgE titer in AP patients reached 1208.1 +/- 421.2 IU/ml in GCF while only 49.1 +/- 50.4 in sera. Soluble CD23 in GCF reached 236.1 +/- 81.3 ng/ml in GCF and 5.6 +/- 1.8 ng/ml in sera. GCF of gingivitis patients, however, contained no detectable sCD23.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7472998 TI - Polyamines found in gingival fluid enhance the secretory and oxidative function of human polymorphonuclear leukocytes in vitro. AB - Many bacterial and host cells contain large amounts of polyamines that can be released at infection sites as a result of cell lysis. Consequently, the putrescine and spermidine content of gingival fluid from inflamed periodontal pockets (0.1 to 1 mM) is sharply elevated in comparison to peripheral blood. At these levels, polyamines potentiated fMet-Leu-Phe-induced Ca2+ signaling in polymorphonuclear leukocytes (PMNs) in vitro. Consistent with the essential role of Ca2+ signaling in PMN activation, secondary granule release and superoxide anion production by fMet-Leu-Phe-stimulated PMNs was enhanced in the presence of polyamines. Thus, polyamines may play a local role in modulating the antimicrobial activity of PMNs in periodontal disease. PMID- 7472999 TI - Detection of antigenic proteins in Porphyromonas gingivalis using two-dimensional electrophoresis and western blots. AB - Using two-dimensional (2D) electrophoresis and western blot assay, we analyzed antigenic proteins in Porphyromonas gingivalis uniquely recognized by antibodies in sera of periodontitis subjects. Proteins in the total membrane fraction of P. gingivalis 381 were resolved into at least 70 protein spots by 2D electrophoresis. In the gel stained with silver, the substance around 47 kDa protein on the acidic side (at an isoelectric point of about 4.5) was stained as a smear. Antigenic substances were characterized using purified IgGs from sera of 16 adult periodontitis (AP), 19 rapidly progressive periodontitis (RPP) and 14 periodontally healthy volunteers. Western blots demonstrated that 75 kDa protein reacted with IgGs from 75% of AP patients (p < 0.001), the antigenic substance around acidic 47 kDa protein reacted with IgGs from 81.3% of AP (p < 0.01) and 68.4% of RPP patients (p < 0.01) and the acidic 47 kDa protein reacted with 87.5% of AP (p < 0.01) and 78.9% of RPP patients (p < 0.01). The reaction frequency was significantly different from that of the healthy volunteers. Also 51 kDa and 41 kDa proteins reacted with 47 and 43 of 49 IgG samples, respectively. The substance around acidic 47 kDa protein reacted with mouse antiserum to P. gingivalis-LPS. After treatment with pronase or heat, the antigenic reactions disappeared not only from the proteins, but also from the area around the acidic 47 kDa protein. When the fraction was digested with lipase, the antigenic reaction of the area decreased.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7473000 TI - Effect of substance P administration on vascular permeability in the rat oral mucosa and sublingual gland. AB - Neuropeptides, including substance P (SP) may play a role in neurogenic inflammation. Although SP-immunoreactive (SP-IR) axons are known to be present within the oral mucosa (OM) and salivary glands, the functional significance of SP in oral mucosa and sublingual salivary gland (SLG) is not fully understood. The present experiments were carried out to study the effects of SP infused into the left common carotid artery on vascular permeability in the OM and in the SLG of male rats. Vascular permeability was assessed on the basis of Evans Blue extravasation. Separate groups of animals received histamine (H1) receptor antagonist (chloropyramine, 10 mg kg-1 i.v.) or prostaglandin synthesis inhibitor (indomethacin, 4 mg kg-1 i.v.) prior to SP infusions. Infusion of SP in doses of 30 and 74 pmol min-1 increased the vascular permeability of OM by 162.3 +/- 16.3% (n = 8, p < 0.05) and 482.7 +/- 46.7% (n = 8, p < 0.001), respectively. SP in a dose of 15 pmol min-1 did not increase Evans Blue extravasation in OM (38.3 +/- 4.0 micrograms g-1, n = 8, compared to the control: 44.0 +/- 7.9 micrograms g-1, n = 8, NS).(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7473001 TI - Effects and interactions of tumour necrosis factor alpha and bradykinin on interleukin-1 production in gingival fibroblasts. AB - Effects of and interactions between tumour necrosis factor alpha (TNF alpha) and bradykinin (BK) on production of interleukin-1 (IL-1 alpha, IL-1 beta) in human gingival fibroblasts were studied. The cytokine TNF alpha induced production of cell-associated IL-1 alpha and IL-1 beta in gingival fibroblasts, with IL-1 beta being most abundant. Addition of BK, in the presence of TNF alpha, for 1 h and 6 h, respectively, synergistically enhanced the TNF alpha induced IL-1 beta production, whereas BK alone did not induce IL-1 production. Similar to BK, two phorbol esters, phorbol 12,13 dibutyrate (PDBu) and phorbol 12-myristate-13 acetate (PMA) which are known to stimulate protein kinase C (PKC), synergistically enhanced the TNF alpha induced IL-1 beta production in the gingival fibroblasts. On the contrary, a phorbol ester which does not activate protein kinase C, 13-phorbolacetate (13-PA), did not potentiate the TNF alpha induced IL-1 beta production. Similar to BK, the phorbol esters (PMA, PDBu, 13 PA) alone did not induce IL-1 beta production in the gingival fibroblasts. The results indicate that TNF alpha induces production of cell-associated IL-1 in gingival fibroblasts, which can be upregulated by a PKC dependent pathway. PMID- 7473003 TI - Influence of sE-selectin and L-selectin on the regulation of cell migration during chronic periodontitis. AB - Soluble endothelial leukocyte adhesion molecule-1 (sE-selectin) levels in peripheral blood (PB) and gingival capillary blood (GCB) of both healthy donors (HD) and patients with adult periodontitis (AP) were assayed by ELISA. Binding of sE-selectin to polymorphonuclear neutrophils (PMN) from PB, GCB and crevicular fluid (GF), and expression of L-selectin and sialyl-Lewisx (sLex) on these cells were analyzed by immunofluorescence and flow cytometry. No significantly enhanced serum levels of sE-selectin in patients with AP, compared to HD (28 +/- 5 ng/ml vs 19 +/- 3 ng/ml, respectively), and no differences in the concentration of sE selectin in GCB (16 +/- 1 ng/ml vs 16 +/- 2 ng/ml, respectively) were observed. On PB-PMN no significant differences in the expression of L-selectin and sLex were found and binding of sE-selectin to PB-PMN was comparable between HD and patients with AP. Binding of sE-selectin to GCB-PMN was significantly higher in patients with AP compared to HD (mean channel fluorescence (MCF) = 88.5 +/- 13.2 vs MCF = 24.2 +/- 5.3, respectively). The expression of sLex on GCB-PMN did not differ significantly between the two groups. A significant decrease in the expression of the adhesion molecule L-selectin on GCB-PMNs compared to PB-PMN was found in patients with AP but not in HD. CF-PMN showed decreased expression of both L-selectin and sLex compared to PMN from PB and GCB, both in HD and patients with AP. Taken together, these data suggest that PMN from patients with AP had reduced selectin-mediated adhesive capabilities to inflamed gingival endothelium. PMID- 7473002 TI - Relationship between oral pain and ethanol concentration in mouthrinses. AB - Previous reports have indicated that certain mouthrinses, even when used as directed can induce oral pain. In order to help determine the causal agent(s), various commercially available mouthrinses, as well as control solutions, were tested in a psychophysical study in which subjects rated categories of pain during and after mouthrinsing. More specifically, the studies tested the effects of ethanol concentration on induced pain. The results show that there is a direct relationship between ethanol content and the amount of induced pain. Furthermore, the amount of pain was found to increase with time of rinsing, and to slowly decrease after cessation of rinsing. Lastly, comparison of ethanol/water controls with a marketed product (Clear Choice) matched for ethanol content showed that, while ethanol was the key factor in mouthwash-induced oral pain, other presently unidentified agents can also add to the effect. PMID- 7473004 TI - Microbial vitality of supragingival dental plaque during initial stages of experimental gingivitis in humans. AB - Although vital plaque micro-organisms are part of the natural ecosystem in the oral cavity they are also the key factor in the development of diseases induced by the human dental plaque. In a previous study (9) the portion of vital bacteria related to the total number of plaque micro-organisms (i.e. the microbial vitality) appeared low in small plaque samples. The objective of this investigation was to determine the exact relationship of microbial vitality and age of supragingival plaque during the early phases of human dental plaque formation. Between intervals of optimal oral hygiene, thirteen participants refrained from all oral hygiene measures for periods of 1, 2, 4, 8, 24 and 72 h. Plaque was completely sampled from a defined area situated on the vestibular surface of the teeth 13, 14, 15, 23, 24 and 25. The pooled plaque from these areas was immediately processed. Total bacterial counts (BC) as enumerated by darkfield microscopy, and colony-forming units (CFU) were recorded. The microbial vitality was calculated indirectly as plating efficiency (PE = CFU per BC) and directly assessed using a vital fluorescence (VF) technique. In the 1 h old plaque samples the median values of PE and VF were 29% and 18%, respectively. Thereafter, the microbial vitality increased significantly with plaque age. The 24 h old plaque samples yielded values of 77% (PE) and 62% (VF). It was concluded that the microbial vitality of the early dental plaque investigated was considerably lower compared to that of a more mature plaque. PMID- 7473005 TI - Development and characterization of an in vitro gingival epithelial model. AB - A 3-dimensional gingival epithelial model has been developed and characterized. Oral epithelial cells and connective tissue fibroblasts were isolated from human gingival tissue and used to create an in vitro oral mucosa co-culture model. Fibroblasts were seeded on a scaffold of nylon mesh, allowed to proliferate and secrete collagen and extracellular matrix proteins to form a stroma capable of supporting the growth of epithelial cells. Epithelial cells were seeded on top of a confluent stromal layer, proliferated and differentiated to form a stratified squamous epithelium. Resident epithelial cells were stimulated, by manipulation of growth medium and culture conditions, to form a multi-layered oral mucosa-like tissue. Histologic analyses revealed cellular architecture exhibiting stromal epithelial interaction which supports the growth and differentiation of an epithelial layer. Immunohistochemical analyses confirmed production of types I and III collagen. Immunofluorescence of the stromal layer identified type IV collagen and fibronectin. Fibronectin was also detected on surface epithelium. Differentiation of basal, spinous and granular cells was observed, and the presence of differentiation markers, acidic (K10, 14-16, 19) and basic (K1-8) cytokeratins were confirmed using broad spectrum cytokeratin antibodies, AE1 and AE3. Development of a discontinuous basal lamina zone, with hemidesmosomes, was observed by electron microscopy. The co-culture was metabolically active, as measured by the thiazoyl blue (MTT) assay for mitchondrial function and [3H] thymidine incorporation into DNA. The human gingival epithelial co-culture model was viable up to 35 days post-epithelial seed. This model may offer opportunities for limited study of periodontal tissue responsiveness. PMID- 7473006 TI - Stimulation of human periodontal ligament fibroblast collagenase production by a gingival epithelial cell-derived factor. AB - To examine whether cell-to-cell interactions between human gingival epithelial cells (HGE) and periodontal ligament fibroblasts (PLF) or gingival fibroblasts (GF) take place in the periodontium, the effects on collagenase production by PLF and GF were analyzed after adding several concentrations of HGE-conditioned medium (HGE-CM) to PLF or GF culture. Collagenase production by both cell populations was stimulated by adding HGE-CM, which stimulated collagenase production by PLF to a greater extent than that by GF. The HGE-derived stimulatory factor had a molecular mass of approximately 20 kDa, and its stimulant effect was inhibited markedly in the presence of an anti-human interleukin-1 alpha (IL-1 alpha) neutralizing antibody, indicating that the factor was identical to, or antigenically cross-reactive with, IL-1 alpha. These results suggest that epithelial apical migration in the periodontium may occur after interstitial resident cells have released tissue-degrading enzymes, such as collagenase, and damaged the extracellular matrix, once a sufficient amount of IL 1 alpha-like factor for stimulating the production of proteolytic enzyme has been released by HGE in periodontal lesions. PMID- 7473007 TI - Understanding diagnostic testing for periodontal diseases. AB - Probability computations are used to characterize new periodontal diagnostic tests and are an integral facet of risk assessment for periodontal diseases. However, misinterpretation of these data can result in confusion and erroneous conclusions regarding the relationship of test results to the presence or absence of disease in a subject, or to episodes of periodontal destruction at specific sites. This paper was written to provide clinicians with a primer to help in understanding calculations used to evaluate diagnostic tests and risk assessment. PMID- 7473008 TI - Gingival interleukin-6 concentration following phase I therapy. AB - There is little information available on the effects of periodontal therapy on tissue levels of interleukin-6 (IL-6). This study compares IL-6 concentrations in gingiva and gingival crevicular fluid (GCF) from patients requiring surgical procedures following Phase I periodontal therapy. Sites requiring surgery due to "unresolved" intrabony pockets were compared to sites not requiring surgery ("resolved") in each patient. Resolved sites had minimal histologic evidence of chronic inflammation and IL-6 accumulation; unresolved sites contained a substantial number of chronic inflammatory cells and IL-6 was widely distributed. IL-6 levels in GCF were significantly greater at resolved than at unresolved gingival sites (P < 0.001); however, IL-6 tissue levels were significantly greater at unresolved than in resolved gingival sites (P < 0.01). These results suggest that IL-6 levels were elevated in gingival connective tissue adjacent to intrabony pockets which had not been resolved following Phase I therapy. Thus, intrabony pocket resolution may be affected by accumulation of IL-6 in the adjacent gingival connective tissue, which may result from increased rates of synthesis or reduced rates of release from gingiva into the GCF. PMID- 7473009 TI - Comparative effectiveness of various regenerative modalities for the treatment of localized juvenile periodontitis. AB - The objective of this study was to assess the effectiveness of surgical debridement versus various guided tissue regeneration techniques for the treatment of localized juvenile periodontitis (LJP). Seven patients, 14 to 18 years old, were selected on the basis of having; 1) vertical bone loss on at least 3 first molars; 2) probing attachment loss > or = 5 mm; 3) presence of Actinobacillus actinomycetecomitans (Aa) as determined by the DNA probe test; 4) no systemic diseases; and 5) not taken antibiotics for 6 months prior to treatment. Each patient was randomly assigned by first molar involvement to one of four treatment modalities: 1) surgical debridement alone (DEBR); 2) ePTFE membrane alone (G); 3) ePTFE membrane plus root conditioning (G + D); or 4) ePTFE membrane plus root conditioning plus composite graft (G + D + B). Standardized clinical attachment level measurements (CAL) were taken at baseline, 6, 12, and 18 months. Hard tissue measurements were evaluated radiographically (RAD) at baseline, 6, 12, and 18 months, and volumetrically at baseline and 12 months post surgery. At 12 months re-entry, all residual defects were re-treated with a composite graft consisting of calcium sulfate + DFDBA + doxycycline. Results were evaluated 6 months post-retreatment using CAL and RAD measurements. Statistical analysis of the data showed: 1) a significant gain in clinical attachment as well as increased volumetric bone fill of the defects 12 and 18 months following the various treatment modalities when compared to baseline (P < 0.05); and 2) no statistically significant differences among the tested treatment modalities and surgical debridement alone for all parameters (P > 0.05). PMID- 7473010 TI - Correlation between volatile sulphur compounds and certain oral health measurements in the general population. AB - Oral malodor was measured using a portable sulphide monitor in 2,672 individuals aged 18 to 64 years. In addition, dental (DMFT) and periodontal conditions (CPITN and attachment loss), dental plaque, and tongue coating status were assessed. Before clinical examination, subjects were interviewed about their oral health habits, smoking habits, and medical history. Data on volatile sulphur compounds (VSC) were analyzed by gender, age group, and time of measurement. There were no significant differences observed in the VSC between males and females in any age group. In each age group, the measured values of oral malodor were highest in the late morning group (58.6 ppb in average), followed by the late afternoon group (52.1 ppb), while lowest values were shown in the early afternoon group (39.4 ppb). Significant correlation was observed only between the VSC value and periodontal conditions and tongue coating status. The results also suggest that oral malodor might be caused mainly by tongue coating in the younger generation and by periodontal diseases together with tongue coating in older cohorts in the general population. Age was not a risk factor for increasing VSC. PMID- 7473011 TI - Effect of tetracycline HCl on periodontally-affected human root surfaces. AB - Scanning electron microscopy was used to evaluate surface characteristics of periodontitis-exposed instrumented human cementum and dentin surfaces following topical application of tetracycline HCl (TTC). Specimens were randomly assigned to application of sterile saline for 1 minute (control); TTC (10 mg/ml) for 1 minute and 4 minutes, respectively; and TTC (100 mg/ml) for 1 minute and 4 minutes, respectively. Solutions were applied with a cotton pellet using a burnishing technique. Control specimens exhibited an amorphous irregular surface smear layer. TTC treatment of cementum for 1 minute resulted in a relatively debris-free, nonhomogeneous surface. The 4-minute application resulted in a surface exhibiting a densely fibrillar, mat-like texture. Dentin specimens conditioned for 1 minute showed a smooth surface with many tubule openings partially occluded by debris. The 4-minute treatment exposed a 3-dimensional network of intertubular and peritubular collagen fibrils. No consistent morphologic differences were observed between cementum or dentin specimens treated with TTC at concentrations of 10 and 100 mg/ml, respectively. The results suggest that topical application of TTC produces morphologic alterations of periodontitis-exposed cementum and dentin that appear related to application interval rather than concentration of the drug. PMID- 7473012 TI - Evaluation of periodontal treatments using controlled-release tetracycline fibers: clinical response. AB - The purpose of this investigation was to evaluate the clinical efficacy of controlled-release tetracycline fiber therapy in adult periodontitis patients. One hundred-twenty-two (122) adult patients from 3 dental centers were enrolled at baseline for this study. each patient provided at least one site in each of four quadrants that was > or = 5 mm and bled on probing. One or two such sites were selected as test sites and were randomly assigned to receive one of four treatments: scaling and root planing (S), scaling and root planing plus tetracycline fiber for 10 days (SF), fiber therapy alone for 10 days (F), or two 10-day serial fiber applications (FF). After treatment, no periodontal maintenance or supportive care was provided until the end of this 12-month study. Probing depth (PD), clinical attachment level (CAL), plaque, and bleeding on probing (BOP) were measured at baseline and at 1, 3, 6, 9, and 12 months following treatment. Repeated PD and CAL measurements were taken at three locations within each site and averaged for each site. One hundred-sixteen (116) subjects completed the study. All treatments resulted in similar improvements in clinical parameters compared to baseline and were equally effective in the treatment of periodontitis as measured by probing depth reduction, clinical attachment level gain, and reduction of bleeding on probing. The clinical response, established primarily by 3 months following therapy, was generally sustained in all treatment groups for 12 months without the benefit of supportive maintenance therapy. PMID- 7473013 TI - Evaluation of periodontal treatments using controlled-release tetracycline fibers: microbiological response. AB - In a 12-month multi-center study of 116 adult periodontitis subjects, six putative periodontal pathogens were monitored by DNA probe methods in a subset of 31 subjects. Monitored species included Porphyromonas gingivalis (Pg), Prevotella intermedia (Pi), Fusobacterium nucleatum (Fn), Eikenella corrodens (Ec), Campylobacter rectus (Cr), and Actinobacillus actinomycetemcomitans (Aa) with an average detection limit of 1.8 x 10(4) bacterial colony forming units/sample. The microbiological response to four periodontal treatments was studied, one treatment in each quadrant; scaling and root planing (S), scaling and root planing with tetracycline (TC) fiber (SF), a single application of TC fiber (F) and two serial applications of TC fiber (FF). Generally two sites were sampled in each quadrant, however, in some quadrants only one site was selected. These treatments were evaluated at baseline; immediately following therapy; and post treatment at 1, 3, 6, and 12 months. The study was conducted with a split-mouth design with no maintenance therapy over a 12-month period. At baseline, 70.8% of sites had detectable Fn; 42.9% Pg; 63.5% Pi; 29.7% Ec; 28.3% Cr; and 5.5% Aa. No significant differences were seen in baseline proportions of these species between centers. Numbers and proportions of detectable pathogens (with the exception of Pg) exhibited a triphasic temporal response: a precipitous initial decrease immediately following therapy; a rise in proportions in the 1- to 3 month post-therapy period; and a spontaneous decline in the absence of therapy over the 3- to 12-month period.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7473014 TI - Evaluation of periodontal treatments using controlled-release tetracycline fibers: maintenance response. AB - The purpose of this investigation was to examine periodontal disease recurrence from 3 to 12 months following various treatments with scaling and root planing and controlled-release tetracycline fibers. One-hundred-twenty-two (122) adult volunteers with at least one bleeding pocket > or = 5 mm in each of four quadrants were enrolled in this study. One or two such sites in each quadrant were selected as test sites. Quadrants were randomly assigned to receive one of four treatments: scaling and root planing (S); scaling and root planing plus tetracycline fiber for 10 days (SF); fiber therapy alone for 10 days (F); or fiber therapy alone for 20 days (FF). After treatment, no supportive care was provided during the 12-month study period. Probing depth (PD), attachment loss (AL), plaque, and bleeding on probing were measured at baseline, and at 1, 3, 6, 9, and 12 months after treatment. PD and AL measures were taken at three locations within each site and averaged for each site. Disease recurrence was defined as > or = 1 mm mean attachment loss at a site during the 3- to 12-month period. One-hundred-sixteen (116) subjects completed the study. Sites treated with SF experienced significantly (P < 0.05) less disease recurrence (4%) than S, F, or FF (9%, 10%, and 12%, respectively). Results of this study suggest that, compared to S, F, or FF, scaling and root planing in conjunction with tetracycline fiber therapy for 10 days can significantly reduce disease recurrence 3 to 12 months following treatment in the absence of supportive care. PMID- 7473016 TI - Eosinophilic granuloma of the mandible involving the periodontal tissues. A case report. AB - This case reports on a 14-year-old boy with multiple radiolucent lesions widely distributed in the mandible, particularly in the right side (body-angle-ascending ramus). The lesion was first diagnosed as early-onset periodontitis and treated accordingly. It continued to grow very quickly, however, over a 6-month period, resulting in involvement of the entire mandible, with complete destruction of the lingual cortical bone, erosion of the external cortex with expansion in the neighboring soft tissues, and with a periosteal reaction in some areas. This case may serve to illustrate the rapid progress of eosinophilic granuloma, the importance and the difficulties of an early diagnosis, and the consequences of late diagnosis and treatment. PMID- 7473015 TI - The origin of fibroblasts and their role in the early stages of horizontal furcation defect healing in the beagle dog. AB - The origin of fibroblasts, their proliferative activity and roles in the early stages of periodontal repair were investigated in order to better understand the periodontal healing process in furcation defects of the beagle dog after guided tissue regenerative therapy. Newly divided cells were identified by immunolocalization of bromodeoxyuridine (BrdU) injected 1 hour prior to sacrificing the animals. At 1 and 2 weeks after creation of the defects, the lesions were occupied primarily by granulation tissue. Under this condition, periodontal ligaments (PDL) fibroblasts in a coronal portion of the remaining PDL close to wounds proliferated actively, migrated along the root surface and formed fibrous connective tissue on the surface. Similarly, the fibroblasts adjacent to the bone surface also showed proliferative activity and engaged in active formation of fibrous connective tissue on the bone surface. The majority of labeled cells in both areas were located in the extravascular area. At 3 and 4 weeks, the defects were filled with an increased amount of new connective tissue and bone. The labeled fibroblasts were preferentially found in the most coronal portion of connective tissue formed on the root surface that was in direct contact with inflamed tissue, and the collagen fibers projected into granulation tissue. In areas of active bone formation, numerous labeled fibroblasts were located in connective tissue adjacent to the newly-formed bone. However, fibroblasts in the endosteum of new bone were rarely labeled These results indicate that fibroblasts involved in periodontal repair originate primarily from both the remaining PDL and alveolar bone, and actively engage in fibrous connective tissue formation in the early stages of periodontal repair The ability of PDL fibroblasts to proliferate, migrate, and form connective tissue on the root surfaces in the early repair stages appears to play a crucial role in the formation of the PDL and cementum, and consequently, in periodontal regeneration in the absence of root resorption and ankylosis. As the formation of new connective tissue and bone continues, the precursor cells for fibroblasts and osteoblasts are supplied locally through the continued divisions of the fibroblastic cells in association with the newly-formed connective tissue. Paravascular and endosteal cells appear to be minor contributors to new cell population during furcation defect repair in the beagle dog. PMID- 7473017 TI - Benign mucous membrane pemphigoid with advanced periodontal involvement: diagnosis and therapy. AB - This paper describes a case of benign mucous membrane pemphigoid which affected the periodontal tissues and complicated ocular prosthesis placement. The clinical findings, diagnostic criteria, and therapeutic management of this disorder are reviewed. PMID- 7473018 TI - Iatrogenic gingival overgrowth in cardiac transplantation. AB - It is well established that both cyclosporin and nifedipine are associated with gingival overgrowth. Although both drugs are widely used in the management of organ transplant patients, there is little information on the prevalence and severity of this unwanted effect in cardiac transplant patients. This study evaluated the gingival health of 94 dentate cardiac transplant patients, all of whom were medicated with cyclosporin as a component of their immunosuppressive therapy. Sixty-three (63) of the patients were also medicated with nifedipine. Significantly higher gingival overgrowth scores (P < 0.0001) and periodontal probing depths (P = 0.001) were observed in patients medicated with the combination of cyclosporin and nifedipine than those medicated with cyclosporin alone. Likewise, there was a significantly greater need to carry out gingival surgery on patients taking the combination (62%), than those medicated with cyclosporin alone (25.8%) (P = 0.001). Patient's age, sex, duration of therapy, gingival bleeding index, and nifedipine therapy were important determinants for both the expression of gingival overgrowth and the need for surgery. Significant sequestration of nifedipine in the gingival crevicular fluid (GCF) was observed. The concentration of nifedipine in GCF did not relate to either the gingival changes or plasma concentration of the drug. Cardiac transplant patients are at risk of developing gingival overgrowth and approximately 50% require surgical intervention. This risk increases significantly when patients are medicated concomitantly with nifedipine. PMID- 7473019 TI - Vesiculo-ulcerative diseases and periodontal practice. PMID- 7473020 TI - Re: Local predisposing events leading to gingivitis and periodontitis. PMID- 7473021 TI - Additivity of nonconscious affect: combined effects of priming and exposure. AB - Affect deriving from 2 independent sources--repeated exposure and affective priming--was induced, and the combined effects were examined. In each of 4 studies, participants were first shown 72 Chinese ideographs in which the frequency of exposure was varied (0, 1, or 3). In the second phase participants rated ideographs that were primed either positively, negatively, or not at all. The 4 studies were identical except that the exposure duration--suboptimal (4 ms) or optimal (1 s)--of both the initial exposure phase and the subsequent priming phase was orthogonally varied. Additivity of affect was obtained only when affective priming was suboptimal, suggesting that nonconscious affect is diffuse. Affect whose source was apparent was more constrained. Interestingly, increases in liking generated through repeated exposures did not differ as a function of exposure duration. PMID- 7473022 TI - When less is more: counterfactual thinking and satisfaction among Olympic medalists. AB - Research on counterfactual thinking has shown that people's emotional responses to events are influenced by their thoughts about "what might have been." The authors extend these findings by documenting a familiar occasion in which those who are objectively better off nonetheless feel worse. In particular, an analysis of the emotional reactions of bronze and silver medalists at the 1992 Summer Olympics--both at the conclusion of their events and on the medal stand- indicates that bronze medalists tend to be happier than silver medalists. The authors attribute these results to the fact that the most compelling counterfactual alternative for the silver medalist is winning the gold, whereas for the bronze medalist it is finishing without a medal. Support for this interpretation was obtained from the 1992 Olympics and the 1994 Empire State Games. The discussion focuses on the implications of endowment and contrast for well being. PMID- 7473024 TI - Agreement among judges of personality: interpersonal relations, similarity, and acquaintanceship. AB - Personality judgments of 184 targets were provided by the self, college acquaintances, hometown acquaintances, parents, and strangers. Study 1 found that knowing the target in the same context enhanced but was not necessary for interjudge agreement and that acquaintances who had never met agreed with each other as well as those who had met. Study 2 found that personality judgements by acquaintances manifested much better interjudge and self-other agreement than did judgments by strangers. Acquaintances were not more similar to their targets than were strangers, and their accuracy derived more from their distinctive judgment of the target than from assumed similarity. These results rule out overlap, communication, and assumed similarity as necessary bases of interjudge agreement and thereby support the simpler hypothesis that interjudge agreement stems from mutual accuracy. PMID- 7473023 TI - Expert role assignment and information sampling during collective recall and decision making. AB - Previous research has found that decision-making groups do not effectively pool unshared information. This study examined how personal expertise facilitates the mentioning and validation of unshared information in collective recall and decision-making groups by increasing members' awareness of who holds what types of information. Assigned expertise increased substantially the proportion of unshared information mentioned during both collective recall and decision-making tasks. Two results supported the hypothesis that assigned expertise provides validation for the recall of unshared information. When expertise was assigned, (a) more of the unshared information mentioned during the recall task was retained on the collectively endorsed written protocol, and (b) unshared information that was mentioned in discussion was more likely to be correctly recognized by members after group interaction. PMID- 7473025 TI - Traits and metatraits: their reliability, stability, and shared genetic influence. AB - Metatraits measure individual differences in construct relevancy, whereas traits measure individual differences in construct extremity. Twenty-four traits and metatraits were examined in this study using 157 pairs of identical twins reared together, 95 pairs of identical twins reared apart, 211 pairs of fraternal twins reared together, and 228 pairs of fraternal twins reared apart obtained from the Swedish Adoption/Twin Study of Aging (see N. L. Pedersen et al., 1991). Reliability and stability analyses of the metatraits revealed generally lower reliability and stability across time compared to traits. Quantitative genetic analyses of the relationship between traits and metatraits indicated that approximately 69% of the metatrait's genetic variance is shared with the trait, with 31% of its genetic variance unique to the metatrait. These results suggest that metatraits provide a useful additional view of personality. PMID- 7473026 TI - Children's action-control beliefs about school performance: how do American children compare with German and Russian children? AB - Using the revised Control, Agency, and Means-ends Interview (T. D. Little, G. Oettingen, & P. B. Baltes, 1995), we compared American children's (Grades 2-6) action-control beliefs about school performance with those of German and Russian children (Los Angeles, n = 657; East Berlin, n = 313; West Berlin, n = 517; Moscow, n = 541). Although we found pronounced cross-setting similarities in the children's everyday causality beliefs about what factors produce school performance, we obtained consistent cross-setting differences in (a) the mean levels of the children's personal agency and control expectancy and (b) the correlational magnitudes between these beliefs and actual school performance. Notably, the American children were at the extremes of the cross-national distributions: (a) they had the highest mean levels of personal agency and control expectancy but (b) the lowest beliefs-performance correlations. Such outcomes indicate that the low beliefs-performance correlations that are frequently obtained in American research appear to be specific to American settings. PMID- 7473027 TI - The structure of psychological well-being revisited. AB - A theoretical model of psychological well-being that encompasses 6 distinct dimensions of wellness (Autonomy, Environmental Mastery, Personal Growth, Positive Relations with Others, Purpose in Life, Self-Acceptance) was tested with data from a nationally representative sample of adults (N = 1,108), aged 25 and older, who participated in telephone interviews. Confirmatory factor analyses provided support for the proposed 6-factor model, with a single second-order super factor. The model was superior in fit over single-factor and other artifactual models. Age and sex differences on the various well-being dimensions replicated prior findings. Comparisons with other frequently used indicators (positive and negative affect, life satisfaction) demonstrated that the latter neglect key aspects of positive functioning emphasized in theories of health and well-being. PMID- 7473028 TI - Individual differences in cardiac sympathetic control predict endocrine and immune responses to acute psychological stress. AB - Potential mechanisms coordinating individual differences in cardiovascular reactivity and endocrine and immune responses to acute psychological stress were examined. Twenty-three young, healthy women performed a mental arithmetic challenge while measures of cardiovascular, endocrine, and immune function were assessed. Results revealed that the acute stressor was associated with changes in the cardiovascular, endocrine, and immune systems. More important analyses revealed that individual differences in cardiovascular reactivity predicted stress-induced cortisol changes. Furthermore, cardiac sympathetic control, as indexed by preejection period, was specifically related to changes in natural killer cell activity. These results suggest that distinct physiological pathways are activated in response to acute psychological stress. PMID- 7473029 TI - The three faces of interpersonal dependency: structural analyses of self-report dependency measures. AB - Using the interpersonal circumplex and the 5-factor model of personality as a structural framework, the authors identified 3 aspects of interpersonal dependency in structural analyses of the universe of content defined by the combined item pool of widely used self-report dependency measures: love dependency, exploitable dependency, and submissive dependency. Markers of these aspects of interpersonal dependency were reconstituted, and their relations with neuroticism (N), conscientiousness (C), and openness to experience (O) were examined. All forms of interpersonal dependency are related positively with N. Exploitable and submissive dependency are increasingly negatively correlated with C and O, whereas love dependency is positively correlated with C and O. Implications regarding the assessment of dependency and multifactor models of dependency are discussed, and hypotheses for future research linking dependency and depression are proposed. PMID- 7473030 TI - Central and peripheral self-conceptions are differentially influenced by mood: tests of the differential sensitivity hypothesis. AB - This article proposes a differential sensitivity hypothesis, according to which central (i.e., relatively high in personal descriptiveness and importance) and peripheral (i.e., relatively low in personal descriptiveness and importance) self conceptions are differentially influenced by mood: Peripheral self-conceptions are subject to a mood-congruency bias, whereas central self-conceptions are unaffected by mood. In 4 experiments, participants were first placed into a sad, neutral, or happy mood state through a guided imagery task and later completed behavior self-descriptiveness ratings, trait self-descriptiveness ratings, and trait self-descriptiveness judgmental latencies. Strong support for the differential sensitivity hypothesis was obtained. Peripheral self-conceptions were influenced by mood because they were less elaborated and consolidated and were held with lower certainty, thus increasing the likelihood for the occurrence of constructive, affect-infusing processes (J.P. Forgas, 1995a). PMID- 7473031 TI - A prospective test of an integrative interpersonal theory of depression: a naturalistic study of college roommates. AB - This study tested an integrated interpersonal theory of depression, which combines J. C. Coyne's (1976b) interpersonal theory of depression with work on the interplay between self-enhancement and self-consistency theory. Students' (targets') and their same-gender roommates' appraisals of each other, depression and anxiety levels, reassurance seeking, and negative feedback seeking were assessed at Time 1 (T1), and again at Time 2 (T2), 3 weeks later. Consistent with the theoretical integration (a) Depressed targets reported engaging in more negative feedback seeking than nondepressed targets, and tended to report seeking more reassurance than nondepressed targets at T1; (b) For male (but not female) targets, the combination of negative feedback seeking, high reassurance seeking, and depression at T1 predicted T1 to T2 increases in rejection by roommates; and (c) Rejection effects applied to depressive symptoms, but not anxious symptoms or anhedonic mood. PMID- 7473032 TI - Stereotype threat and the intellectual test performance of African Americans. AB - Stereotype threat is being at risk of confirming, as self-characteristic, a negative stereotype about one's group. Studies 1 and 2 varied the stereotype vulnerability of Black participants taking a difficult verbal test by varying whether or not their performance was ostensibly diagnostic of ability, and thus, whether or not they were at risk of fulfilling the racial stereotype about their intellectual ability. Reflecting the pressure of this vulnerability, Blacks underperformed in relation to Whites in the ability-diagnostic condition but not in the nondiagnostic condition (with Scholastic Aptitude Tests controlled). Study 3 validated that ability-diagnosticity cognitively activated the racial stereotype in these participants and motivated them not to conform to it, or to be judged by it. Study 4 showed that mere salience of the stereotype could impair Blacks' performance even when the test was not ability diagnostic. The role of stereotype vulnerability in the standardized test performance of ability stigmatized groups is discussed. PMID- 7473033 TI - Subtyping versus subgrouping processes in stereotype representation. AB - Participants were instructed to organize information about group members either by distinguishing stereotype-consistent from stereotype-inconsistent individuals (subtyping instructions), by dividing the individuals into multiple groups on the basis of similarities and differences (subgrouping instructions), or with no explicit organizing instructions. Participants given the subtyping instructions showed greater perceived stereotypicality and homogeneity and perceived a greater difference in how typical the confirming versus disconfirming group members were, relative to subgroup participants. Study 2 demonstrated natural variation among participants in the perceived atypicality of the disconfirming relative to confirming individuals when learning about a gay activist group. Atypicality predicted perceptions of this group, even when prejudice and strength of stereotyping toward gays as a whole were statistically controlled. PMID- 7473034 TI - Change in job and marital experiences and change in psychological distress: a longitudinal study of dual-earner couples. AB - Are changes in job quality more closely linked to changes in distress for men than for women? Conversely, are changes in marital quality more closely linked to changes in distress for women than for men? These questions were addressed in a longitudinal analysis of a random sample of 210 full-time employed dual-earner couples. Change over time in job role quality was significantly associated with change over time in distress, and the magnitude of the relationship differed little, if at all, by gender. In contrast, change over time in marital role quality was also associated with change in distress, but the magnitude of the association depended on gender. Among full-time employed married women, change in marital experience was more closely linked to change in distress than among their husbands. PMID- 7473035 TI - Factors predicting the subjective well-being of nations. AB - Subjective well-being (SWB) in 55 nations, reported in probability surveys and a large college student sample, was correlated with social, economic, and cultural characteristics of the nations. The SWB surveys, representing nations that include three fourths of the earth's population, showed strong convergence. Separate measures of the predictor variables also converged and formed scales with high reliability, with the exception of the comparison variables. High income, individualism, human rights, and societal equality correlated strongly with each other, and with SWB across surveys. Income correlated with SWB even after basic need fulfillment was controlled. Only individualism persistently correlated with SWB when other predictors were controlled. Cultural homogeneity, income growth, and income comparison showed either low or inconsistent relations with SWB. PMID- 7473036 TI - A framework for studying personality in the stress process. AB - This article presents a framework for studying personality in the stress process. The framework specifies that personality may affect both exposure and reactivity to stressful events and that both processes may explain how personality affects health and psychological outcomes. The framework also specifies that personality differences in reactivity may be due to differential choice of coping efforts and differential effectiveness of those efforts. In a 14-day daily study of 94 students, this framework was used to analyze the links among neuroticism, daily interpersonal conflicts, coping with conflicts, and distress. Results showed that high-neuroticism participants had greater exposure and reactivity to conflicts. Furthermore, high- and low-neuroticism participants differed both in their choice of coping efforts and in the effectiveness of those efforts, a possibility not considered in previous models of personality in the stress process. PMID- 7473037 TI - Racial identity, academic achievement, and the psychological well-being of economically disadvantaged adolescents. AB - The relation between racial identity and personal psychological functioning was examined within the framework of the "racelessness" construct proposed by Fordham and Ogbu (S. Fordham, 1988; S. Fordham & J. U. Ogbu, 1986). These researchers have proposed that academically successful African American students achieve their success by adopting behaviors and attitudes that distance them from their culture of origin, resulting in increased feelings of depression, anxiety, and identity confusion. Studies 1 and 2 describe the development of the Racelessness Scale (RS) designed to test these assumptions. Study 2 also investigated Race X Achievement level differences in students' responses to the RS. In Study 3, correlations between the RS and measures of depression, self-efficacy, anxiety, alienation, and collective self-esteem were assessed. The pattern of results in Study 2 suggest that the behaviors and attitudes described by Fordham and Ogbu are common to high-achieving adolescents and not specific to African Americans. However, racial differences in the pattern of associations between the RS and measures of depression suggest that racelessness may have important psychological consequences for African American adolescents. PMID- 7473038 TI - Culture, gender, and self: a perspective from individualism-collectivism research. AB - Individualism and collectivism are often equated with independent vs. interdependent, agentic vs. communal, and separate vs. relational self construals. Although these same concepts have been used to characterize both cultural and gender differences, a perspective of cultural evolution suggests it is unlikely. A division of labor within society may produce gender differences, but this cannot explain cultural differences. A study of self-construal involving 5 cultures (Australia, the United States, Hawaii, Japan, and Korea) shows that differences between these cultures are captured mostly by the extent to which people see themselves as acting as independent agents, whereas gender differences are best summarized by the extent to which people regard themselves as emotionally related to others. PMID- 7473039 TI - Assessing the interpersonal transaction cycle: convergence of action and reaction interpersonal circumplex measures. AB - Contemporary interpersonal theory uses 2 models to represent the process and content of interpersonal transactions. The interpersonal transaction cycle is a dynamic model demonstrating the interaction between the overt behaviors and the covert intrapsychic experiences of 2 interacting individuals in action-reaction event chains. The interpersonal circle is a static model codifying individual differences across various interpersonal content domains (e.g., traits, acts, problems, impacts). The present study used 150 pairs of undergraduates to demonstrate cross-observer structural convergence of 2 separate interpersonal circumplex inventories measuring the "action" and "reaction" components of the transaction cycle, providing evidence that both components of the cycle can be captured by existing circumplex measures, and allowing researchers to proceed on to more comprehensive tests of interpersonal theory. PMID- 7473040 TI - Moderating role of trait aggressiveness in the effects of violent media on aggression. AB - Three studies were conducted to test the hypothesis that high trait aggressive individuals are more affected by violent media than are low trait aggressive individuals. In Study 1, participants read film descriptions and then chose a film to watch. High trait aggressive individuals were more likely to choose a violent film to watch than were low trait aggressive individuals. In Study 2, participants reported their mood before and after the showing of a violet or nonviolent videotape. High trait aggressive individuals felt more angry after viewing the violent videotape than did low trait aggressive individuals. In Study 3, participants first viewed either a violent or a nonviolent videotape and then competed with an "opponent" on a reaction time task in which the loser received a blast of unpleasant noise. Videotape violence was more likely to increase aggression in high trait aggressive individuals than in low trait aggressive individuals. PMID- 7473041 TI - Convergence between self and peer in the response-time processing of trait relevant information. AB - Research on "self-schemas" has extensively documented the effects of these hypothesized cognitive structures on the processing of self-knowledge, but it has neglected to establish whether these structures correspond to knowledge other persons possess about an individual's personality traits and whether this peer knowledge is represented by cognitive structures similar to self-schemas. In the present study, "schematic" measures such as response time in self-ratings were compared with traits attributed to participants by well-acquainted peers. Peer ratings were nearly as accurate as self-ratings in predicting response-time patterns in self-description. Moreover, extreme self-ratings by the target individual were predictive of extreme peer-ratings and quick peer-judgments for the trait in question. This pattern of convergence did not depend on any similarity between the peer's own self-ratings and those of the target. These results indicate that the content and processing characteristics of knowledge structures developed of individuals by well-acquainted peers are remarkably similar to the content and properties of the individual's own self-schemas. PMID- 7473042 TI - When avoiding unpleasant emotions might not be such a bad thing: verbal-autonomic response dissociation and midlife conjugal bereavement. AB - It has been widely assumed that emotional avoidance during bereavement leads to either prolonged grief, delayed grief, or delayed somatic symptoms. To test this view, as well as a contrasting adaptive hypothesis, emotional avoidance was measured 6 months after a conjugal loss as negative verbal-autonomic response dissociation (low self-rated negative emotion coupled with heightened cardiovascular activity) and compared with grief measured at 6 and 14 months. The negative dissociation score evidenced reliability and validity but did not evidence the assumed link to severe grief. Rather, consistent with the adaptive hypothesis, negative dissociation at 6 months was associated with minimal grief symptoms across 14 months. Negative dissociation scores were also linked to initially high levels of somatic symptoms, which dropped to a low level by 14 months. Possible explanations for the initial cost and long-term adaptive quality of emotional avoidance during bereavement, as well as implications and limitations of the findings, are discussed. PMID- 7473043 TI - Drinking to regulate positive and negative emotions: a motivational model of alcohol use. AB - The present study proposed and tested a motivational model of alcohol use in which people are hypothesized to use alcohol to regulate both positive and negative emotions. Two central premises underpin this model: (a) that enhancement and coping motives for alcohol use are proximal determinants of alcohol use and abuse through which the influence of expectancies, emotions, and other individual differences are mediated and (b) that enhancement and coping motives represent phenomenologically distinct behaviors having both unique antecedents and consequences. This model was tested in 2 random samples (1 of adults, 1 of adolescents) using a combination of moderated regression and path analysis corrected for measurement error. Results revealed strong support for the hypothesized model in both samples and indicate the importance of distinguishing psychological motives for alcohol use. PMID- 7473044 TI - [Immunohistochemical analysis of adhesion molecules in directional coronary atherectomy specimens]. AB - Chronic inflammatory cells are key components in the progression of atherosclerotic plaques and restenosis after coronary angioplasty. Adhesion molecules are fundamental in inflammatory processes. Therefore, the distributions of intercellular adhesion molecule-1 (ICAM-1) and vascular cell adhesion molecule (VCAM) were investigated in directional coronary atherectomy specimens obtained from 14 patients, in 6 with acute coronary syndromes (myocardial infarction and unstable angina within 1 month), 6 with old myocardial infarction and 2 with stable effort angina. There were eight primary lesions and six restenotic lesions. Atherectomy tissue fragments were snap frozen and cut into 4 microns thick cryostat sections for immunohistochemical staining by avidin-biotin complex immunoperoxidase techniques using adhesion molecule specific monoclonal antibodies BBIG-I1 (ICAM-1) and BBIG-V1 (VCAM). The cells of lesions were characterized in sequential sections by macrophage marker KP1 (CD68), endothelial marker JC/70A (CD31), and smooth muscle cell marker 1A4 (alpha-smooth muscle actin). Four restenotic lesions that had undergone a prior balloon angioplasty within a few months consisted of intimal proliferation and the other lesions were atherosclerotic plaque. Macrophage-rich areas were seen in the lesions from acute coronary syndromes and/or early restenotic lesions. Expression of ICAM-1 or VCAM was strongly associated with macrophage-rich areas, but VCAM staining was weaker than ICAM-1 except in one restenotic lesion. Macrophages that express ICAM-1 and/or VCAM may be important in the unstable plaques and restenotic lesions related to disease activity of ischemic heart disease. PMID- 7473045 TI - [Electrocardiographic characteristics of patients with left circumflex-related myocardial infarction in the acute phase without tented T waves or definite ST elevation]. AB - Acute myocardial infarction (AMI) causing ST depression and T wave inversion has been diagnosed as subendocardial or non-Q myocardial infarction. However, some patients eventually develop strictly posterior infarction with a lesion of the left circumflex coronary artery (LCX). This study attempted to determine the electrocardiographic (ECG) characteristics of such myocardial infarction in 32 patients with definite AMI in whom ECG showed no hyperacute T waves or ST elevation and the LCX was an infarct-related coronary artery. ECG on admission (< 6 hours from the onset), at 24 hours, and on the 14th day were analyzed to evaluate QRS, ST, T, and U waves using calipers and magnifying lens. Sixty-six patients with normal circulation served as controls. The characteristic ECG findings on admission were ST depression in chest leads, and prominent positive U waves in leads V2 or V3 with relatively small T waves. Based on these results we proposed new ECG criteria: 1) ST depression > or = 0.1 mV in 2 consecutive chest leads, 2) prominent positive U wave > or = 0.1 mV in leads V2 or V3, 3) T/U ratio in leads V2 or V3 < or = 4. Considering two of the above criteria as positive, the sensitivity was 71.9%, the specificity 97.0%, and the diagnostic accuracy 88.8%. In 85.2% of the patients, ST depression returned to the baseline by 24 hours. As the amplitude of the U waves decrease gradually, the T/U ratio increased. The R/S ratio in leads V1 or V2 became > or = 1 by 24 hours in 46.4% and the amplitude of R wave in lead V1 increased gradually. T waves in the right precordial leads increased with time. These findings were consistent with isolated strictly posterior myocardial infarction. From these results we identified new ECG criteria: 1) R/S ratio in leads V1 or V2 > or = 1, 2) R wave > or = 0.7 mV in lead V1, 3) T wave > or = 0.5 mV in lead V1. Considering any of the above criteria as positive, the sensitivity was 72.0%, the specificity 87.9%, and the diagnostic accuracy 86.7% on the 14th day. These new ECG criteria of strictly posterior myocardial infarction with the LCX as an infarct-related coronary artery apply at less than 6 hours or at 24 hours from the onset of the symptoms. PMID- 7473046 TI - Circadian variation in baroreflex sensitivity evaluated by beat-to-beat hemodynamic change in patients with essential hypertension. AB - The mechanisms underlying the higher incidence of cardiovascular events in the morning were investigated by studying the circadian variation in hemodynamics and baroreflex sensitivity (BRS) in 20 untreated inpatients with essential hypertension. Direct blood pressure (BP) and heart rate (HR) were recorded using telemetry. Cardiac output (CO) was measured by the dye dilution method. Beat-to beat stroke volume (SV) and total peripheral vascular resistance (TPR) were obtained using the pulse contour method. The coefficient of regression between HR and systolic BP (SBP) change (delta HR/delta SBP = Ahr) was calculated for in 5 consecutive heart beats during which BP decreased spontaneously and linearly (r > 0.9). Similarly, the delta SV/delta SBP (= Asv) and delta TPR/delta SBP (= Atpr) were also measured, and the negative values of these coefficients (-Ahr, -Asv and -Atpr) were calculated. Comparisons between morning (6-11 a.m.) and evening (4-9 p.m.) values showed no significant difference in mean BP (122 vs 127 mmHg) and HR (72 vs 73 bpm). However, CO (3.7 vs 4.2 l/min), -Ahr (0.28 vs 0.43 bpm/mmHg) and Asv (-1.5 vs 1.4 ml/mmHg) were lower in the morning than in the evening (p < 0.01). In contrast, TPR (40 vs 34 mmHg/l/min) and -Atpr (1.2 vs -1.4 min/l) were higher in the morning than in the evening (p < 0.01). These findings suggest that lower -Ahr and -Asv and higher TPR and -Atpr may cause stress to the cardiovascular system in the morning in patients with essential hypertension. PMID- 7473047 TI - [Isoproterenol induces ST-segment elevation without wall motion aggravation and myocardial ischemia in experimental myocardial infarction]. AB - The mechanism of exercise-induced ST-segment elevation in previous myocardial infarction was investigated using isoproterenol infusion with atrial pacing to simulate exercise hemodynamics in seven closed chest dogs with myocardial infarction. The myocardial infarction was induced by cellulose embolization of the left anterior descending artery. One week after infarction, 1) isoproterenol 0.3 microgram/min infusion, 2) right atrial pacing alone, and 3) isoproterenol infusion after the injection of propranolol 0.5 mg/kg were performed. The heart rate was kept constant by atrial pacing during the examination. Precordial ST segment deviation, left ventricular global and regional wall motions, lactate extraction ratio, and regional myocardial blood flow were measured. Isoproterenol induced a significant elevation of the ST-segment, i.e., the sum of ST-segment elevation at chest leads V2, V3 and V4, sigma STV234, increased from 0.32 +/- 0.11 (mean +/- SEM) to 0.82 +/- 0.22 mV (p < 0.01). However, pacing alone and isoproterenol with pretreated propranolol did not. During isoproterenol-induced ST-segment elevation, left ventricular ejection fraction increased (29.2 +/- 3.3 to 39.1 +/- 3.9%, p < 0.01), lactate extraction ratio decreased but within the normal range for lactate metabolism (30.2 +/- 7.0 to 16.7 +/- 4.2%, p < 0.01), and the inner/outer ratio of regional blood flow did not change significantly (0.81 +/- 0.18 to 0.97 +/- 0.13 in the non-infarct border area, NS; 0.37 +/- 0.04 to 0.42 +/- 0.07 in the infarct border area, NS). Isoproterenol enhanced ventricular wall motion in the border and non-infarct areas, but did not induce aggravation in the infarct area.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7473048 TI - [The relationship between the improvement of cardiac function and the myocardial uptake of I-123 metaiodobenzylguanidine in patients with dilated cardiomyopathy treated by beta-blocker]. AB - Chronic beta-blocker therapy improves hemodynamics and cardiac function in patients with idiopathic dilated cardiomyopathy. However, the change in myocardial uptake of I-123 metaiodobenzylguanidine (123I-MIBG) before and after treatment has not been determined. Myocardial imaging with 123I-MIBG was performed before and 2 or 3 months after beta-blocker (bisoprolol) therapy in 11 patients with dilated cardiomyopathy. The following parameters were compared before and after the treatment: 1) New York Heart Association functional class, 2) X-ray cardiothoracic ratio, 3) heart rate and blood pressure, 4) echocardiographic data (left ventricular end-diastolic and end-systolic diameters, and left ventricular ejection fraction), 5) plasma concentrations of epinephrine, norepinephrine and human atrial natriuretic peptide (HANP), and 6) exercise tolerance time by treadmill. The heart-to-mediastinum ratio of 123I-MIBG activities obtained 3 hours after intravenous injection (late H/M) and washout rate improved significantly after beta-blocker therapy. Cardiothoracic ratio, heart rate, echocardiographic parameters, HANP and exercise tolerance also improved significantly. Late H/M had no significant relationship with any of the clinical parameters, but washout rate was significantly related to left ventricular ejection fraction. These findings suggest that washout rate may be useful to assess the effect of short-term beta-blocker therapy in dilated cardiomyopathy patients. PMID- 7473049 TI - [Cardiac dysrhythmias related to extracorporeal shock wave lithotripsy using a piezoelectric lithotriptor in patients with kidney stones]. AB - Shock waves delivered by a spark-gap generator during extracorporeal shock wave lithotripsy (ESWL) for kidney stones frequently cause cardiac arrhythmias. However, detailed data about the arrythmogenicity of this therapy using a piezoelectric lithotriptor are not available even though this type of machine is generally not considered to induce cardiac arrhythmias. The prevalence and nature of cardiac dysrhythmias during piezoelectric ESWL were assessed in 34 patients with kidney stones in whom cardiac disease other than dysrhythmias had been excluded. Each patient underwent 24-hour ambulatory electrocardiography (ECG) recording 2 to 3 days before ESWL. On the day of ESWL, ECG was performed continuously from 4 hours before and throughout the procedure. The relationship between ESWL-related dysrhythmias and autonomic neural activity was also evaluated by heart rate spectral analysis. Piezoelectric ESWL was found to elicit new or worsened tachyarrhythmia originating from the atria or ventricles or both in 20 patients (59%), atrial premature contractions in 4, ventricular premature contractions in 7, both types in 7, atrial and ventricular premature contractions plus extreme sinus bradycardia in 1, and neurally mediated sinus bradycardia and hypotension in 1. However, piezoelectric ESWL-related dysrhythmias were not related to the dysrhythmias seen in the usual life. Heart rate spectral analysis suggested that changes in autonomic neural activity were involved in the mechanism(s) of ESWL-related dysrhythmia. Although lethal dysrhythmias have not yet been encountered at our institution, patients at risk of sustained life threatening cardiac dysrhythmias should be monitored closely even if ESWL for kidney stones is undertaken using a piezoelectric lithotriptor. PMID- 7473050 TI - [Greenfield filter placement following successful cardiopulmonary resuscitation in a patient with sudden death due to pulmonary embolization]. AB - A 54-year-old man was admitted as an emergency following an episode of syncope and severe dyspnea. The clinical presentation and laboratory data strongly suggested pulmonary embolism, so intravenous thrombolytic therapy was performed. Immediately following the initiation of thrombolytic therapy, the patient experienced cardiopulmonary arrest, but he was successfully resuscitated. Ascending venography of the left lower limb demonstrated a large intraluminal thrombus which was apparently floating between the femoral vein and inferior vena cava. In addition to the anticoagulation therapy, a Greenfield filter was implaced intravenously to prevent recurrent embolization. PMID- 7473051 TI - Formation and elution of toxic compounds from sterilized medical products: methylenedianiline formation in polyurethane. AB - The formation of a toxic and carcinogenic compound, methylenedianiline (MDA), in sterilized medical use polyurethane (PU) is discussed. Due to good biocompatibility and biostability, PU is widely used for blood-containing devices. There are two types of PU currently available for medical use. One is chain-extended thermoplastic PU, the other is thermosetting PU used for potting material connecting fibers and modules in artificial dialyzers and plasma separators. Both gamma-ray irradiation and autoclave sterilization are predominantly used for the sterilization of these devices. MDA formation in sterilized PUs by gamma-ray irradiation and by autoclave treatment is compared. The Delany clause in the USA prohibits the manufacture of medical devices producing any toxic compound during fabrication and sterilization, therefore, the formation and elution of MDA and other toxic compounds should be seriously considered. Although MDA formation at a concentration of a few to a few hundred ppb in autoclaved chain-extended thermoplastic PU has been reported, there have been no papers describing MDA formation in autoclaved thermosetting PU potting material, or describing MDA formation in gamma-ray irradiated chain-extended thermoplastic PU and thermosetting PU. We elected to determine whether MDA was in fact produced in Pus sterilized by gamma-ray irradiation or by autoclave sterilization. Our objective was to estimate the risk factor to the human patients or recipient. Our conclusion is to confirm which sterilization of gamma ray or autoclave is more appropriate. No formation of MDA was observed in autoclaved thermosetting PU potting material at 121 degrees C for 60 min. A few ppm of MDA was formed in irradiated potting material. MDA formation increased with increasing irradiation doses. MDA formed in irradiated potting material at 2.5 Mrad (less than one ppm) is not a significant risk to the recipient. The estimated cancer causing risk factor when absorbing one mg MDA/kg body weight/day is 0.29. Twenty-nine persons per 100 persons exhibit a cancer potential. More importantly, our experimental evidence indicated that serum extracts from irradiated potting material contained unknown compounds other than MDA. The presence of these compounds is considerably more significant and toxic. When tested, these compounds proved to be mutagenic in the absence of metabolic activity, thus promoting a need to identify and study further. Compounds indicating mutagenicity increased with increasing irradiation. Small amounts of mutagenic compounds were determined in extract from autoclaved potting material, smaller than that by gamma-ray irradiation. Therefore, autoclave sterilization is more appropriate, providing materials can withstand the process.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 7473052 TI - A review of the effects of gamma radiation on pharmaceutical materials. PMID- 7473053 TI - Sterilization and reprocessing of materials and medical devices--reusability. AB - Problems associated with reprocessing of disposable medical devices such as hemodialysers with resterilization for reuse and changes in material properties with resterilization of polymeric (PVC, polypropylene, polyester, polycarbonate) materials intended for development of disposable devices are reviewed. Reprocessing of hospital supplies, polystyrene microtiter plate and angiographic catheter for reuse is also discussed. PMID- 7473054 TI - [Stereochemistry of highly selective transition metal or Lewis acid-catalyzed asymmetric reactions]. AB - Recent progress of our studies on catalytic asymmetric synthesis is reviewed focused on transition metal-catalyzed intramolecular asymmetric reactions using chiral organo-sulfur groups as main chiral auxiliaries such as asymmetric vinyl cyclopropane-cyclopentene rearrangements and intramolecular asymmetric metallo ene reactions, Lewis acid-catalyzed intramolecular asymmetric ene reactions with chiral sulfinyl groups, and palladium-catalyzed intramolecular asymmetric allylations via chiral enamines. PMID- 7473055 TI - [Structure and function of IL-5 receptor]. AB - Interleukin-5 (IL-5) regulates the production and function of B cells, eosinophils and basophils. In particular, IL-5 plays a critical role in the development of CD5-positive B (B-1) cells. The pleiotropic activity of IL-5 on target cells is directly dependent on the initial binding to IL-5 specific cell surface receptor (IL-5R). The IL-5 signals are mediated through the high affinity IL-5R which is composed of two different polypeptide chains, alpha and beta. The alpha chain is a membrane-penetrated glycoprotein that specifically binds IL-5 and retains features common to the cytokine receptor superfamily. The beta chain by itself does not bind IL-5, but it can convert the low affinity IL-5R into the high affinity IL-5R and in indispensable for IL-5 signal transduction. The beta chain is shared among receptors for IL-5, IL-3 and GM-CSF and is called beta c. The cytoplasmic comains of both IL-5R alpha and beta c are essential for signal transduction. The membrane proximal proline-rich sequence of the cytoplasmic domain of IL-5R alpha was found to be essential for the IL-5-induced proliferative response, expression of nuclear proto-oncogenes such as c-jun, c fos and c-myc, and activation of Bruton's tyrosine and JAK2 kinases. Furthermore, JAK2 activation correlates with proline residues in Pro-Pro-X-Pro motif in the cytoplasmic domain of IL-5R alpha. These results indicate that activation of JAK2 and its substrate is critical to coupling IL-5-induced tyrosine phosphorylation and ultimately mitogenesis. I will discuss about molecular mechanisms of IL-5 signaling and B cell defect in X-linked immunodeficient mice in relation to IL-5 signaling. PMID- 7473056 TI - [New fluorogenic dienophile: synthesis, reaction with vitamin D, vitamin A and microcystins, and application to fluorometric assays]. AB - We designed and synthesized a new sensitive and highly reactive fluorogenic reagent (1a, DMEQ-TAD) targeting a conjugated diene. DMEQ-TAD reacted quantitatively with major vitamin D metabolites and synthetic analogs under a variety of conditions to yield the corresponding 6,19-cycloadducts as a mixture of the C(6) epimers. The stereochemistry of the adducts was determined by their CD spectra on the basis of the exciton chirality method. The fluorescent products can be quantified linearly down to 10 fmol by HPLC. The new fluorometric method was successfully used in the assays of 25-OHD3, 24,25-(OH)2D3 and 25,26-(OH)2D3 in the human plasma. The method was proved to be reliable and precise compared with the HPLC-UV assay method. Reactions of DMEQ-TAD with retinoic acid (21a) and its geometrical isomers (21b-d) having a conjugated pentaene system afforded 7,10 adducts (24a-d) as major products together with 5,8-adducts (26a-d) in about 9:1 ratio (70-95% yield) except for the 9Z-isomer (21e) which gave only a 5,8,11,14 bis-adduct (27). DMEQ-TAD reacted with microcystins LR, YR and RR, at the conjugated diene part to yield 4,7-cycloadducts as a pair of epimers in good yield. PMID- 7473057 TI - [Synthesis and antiplatelet activity of 2-amino-4,5-diphenylthiazole derivatives]. AB - A series of 4,5-diphenylthiazoles containing morpholinoalkyl amino groups at the 2 position was synthesized and their anti-platelet activities were evaluated. The inhibitory effects of the compounds in which phenyl groups at the 4 position were substituted by methoxy groups were more potent than those of aspirin and ibuprofen on the collagen-induced rabbit platelet aggregation. These compounds were also shown to have potent efficacies on the arachidonic acid-induced platelet aggregation. On the other hand, these compounds did not affect the ADP induced platelet aggregation similarly to non-steroidal anti-inflammatory agents. Moreover, their efficacies on the inhibition of prostaglandin synthesis were more potent than that of ibuprofen. Therefore, we thought that 2 (morpholinoalkyl)amino-4,5-diphenylthiazoles, in spite of their basic properties, inhibited the platelet aggregation based on cyclooxygenase. PMID- 7473058 TI - [Studies on attenuation of post-ischemic brain injury by kampo medicines inhibitory effects of free radical production. II]. AB - It is thought that highly reactive oxygen radicals generated at the ischemia reperfusion in case of strokes play an important role in damaging the brain. It is well known that lipid peroxidation is propagated by active oxygen radicals, and the the brain is susceptible to the lipid peroxidation. In the previous study, we found that several Chinese herbal medicines and kampo components, which were used for the attenuation of the post-ischemic brain injury, showed a free radical (OH., O2-. and DPPH) scavenging activity. However, it is not clear whether these Chinese herbal medicines can inhibit the lipid peroxidation reaction or not. In attempting to address this question, we have used three kinds of kampo formulations (Oren-gedoku-to (Huang-Lian-Jie-Du-Tang), Saiko-ka-ryukotsu borei-to (Chai-Hu-Jia-Long-Gu-Mu-Li-Tang) and Keishi-bukuryo-gan (Gui-Zhi-Fu-Ling Wan)) to measure the suppressive effect of the lipid peroxidation on the mouse cerebrum using the TBA technique in vitro and in vivo. In vitro experiments, all these Chinese herbal medicines decreased the levels of TBA-reactive substances concentration-dependently. In vivo studies, the levels of the TBA-reactive substance of the cerebrum homogenate of mice treated with these kampo formulations by p.o. for three weeks also decreased. From these results, we suggest that the pharmacological action of Chinese herbal medicines used for the attenuation of the post-ischemic brain damage not only have a free-radical scavenging activity, but also have a suppressive effect on the generation of the lipid peroxidation. PMID- 7473059 TI - [Study on baths with crude drug. II.: the effects of coptidis rhizoma extracts as skin permeation enhancer]. AB - Skin permeation and its enhancing activity of Coptidis Rhizoma (Coptis japonica Makino) were studied in regard to its application as a bath agent. As a result, methanol extracts and three alkaloids (berberine, coptisine, and palmatine isolated from Coptidis Rhizoma) enhanced effectively the skin permeation of 5 fluorouracil, which is taken as a hydrophilic permeant. Furthermore, it was observed that diffusion coefficient is almost constant on the skin permeation of 5-fluorouracil and was also observed that these three kinds of alkaloids did not penetrate through the skin, but adsorbed into the skin. These results suggest that these three protoberberine type alkaloids increase the concentration of polar drugs in the skin and enhance the skin permeation similarly to surfactants. PMID- 7473060 TI - [Inhibitory effects of hydroquinone-alpha-glucoside on melanin synthesis]. AB - Inhibitory effects of hydroquinone-alpha-glucoside (HQ-alpha-G) on the melanogenesis were investigated and compared with those of arbutin. The levels of inhibitory effects of HQ-alpha-G and arbutin on the tyrosinase activity were nearly the same. Inhibitory effects of both compounds on the melanogenesis, were studied using cultured B16 melanoma cells, and HQ-alpha-G was also found to have a similar effect to that of arbutin without inhibiting cell growth. In this experiment, while HQ-alpha-G hardly inhibited cell growth at 1 mM, arbutin inhibit it significantly at the same concentration. From these results it is suggested that HQ-alpha-G as well as arbutin inhibited the melanogenesis by affecting tyrosinase rather than by killing melanocytes. Furthermore, the melanogenesis of guinea-pigs with brown hair was reduced to about 80% by applying them each compound. The great differences in toxicity to normal human keratinocyte were not recognized between these two glucosides. It is, therefore, considered that HQ-alpha-G is an effective and safe ingredient for cosmetics. PMID- 7473061 TI - [Stereoselective N-demethylation of chlorpheniramine in rat liver microsomes: studies on age and sex differences]. AB - We have established a simple method for the chiral stationary-phase liquid chromatographic resolution of racemic chlorpheniramine (Chp) and its two N demethylated metabolites, monodesmethylchlorpheniramine (DMChp) and didesmethylchlorpheniramine (DDMChp), using an ovomucoid-conjugated column with respective quantitation limits of 5 ng/ml. The assay was used to study the age and sex difference in stereoselective N-demethylation of Chp by rat liver microsomes. The formation rate of each DMChp from racemic Chp was about 2.2 times faster with the S-(+)-enantiomer than the R-(-)-enantiomer in male rats at the age of only 8 weeks, whereas the rate of N-demethylation in male rats was not different between S-(+)- and R-(-)-enantiomers at the age of 3 and 24 weeks. The Vmax/Km value for the formation of S-(+)-DMChp increased with age in male rats, but the value in 8-week-old rats was lower than that in 3-week-old rats in the formation of R-(-)-DMChp. In female rats, on the other hand, the formation rate of DMChp from racemic Chp did not differ between S-(+)- and R-(-)-enantiomers at each age, suggesting a lack of stereoselectivity in the microsomal N demethylation. Moreover, the Km and Vmax values for N-demethylation of Chp enantiomers were nearly identical between two consecutive ages in female rats. Further metabolism of DMChp to DDMChp was not observed between both enantiomers in male and female liver microsomes at the age of 3, 8 and 24 weeks. PMID- 7473062 TI - [Effect of changes in the lipid composition on the plasma membrane of Saccharomyces cerevisiae through mutation of the phase transition and mixing behavior of the lipid fraction]. AB - Mutation of Saccharomyces cerevisiae leads to an alteration of the size and surface structure of the mutant cell. These phenomena are correlated with change in the lipid composition and hence membrane fluidity of the plasma membranes. Such alterations are in the fatty acyl constituents of phospholipids and glycolipids and include changes in the saturation or length of fatty acyl chains. Simultaneously, the "melting point" of phospholipid fractions, i.e. the temperature of the ordered-disordered phase transition and mixing behavior, changed. Consequently calorimetry on isolated phospholipid fractions extracted from plasma membranes is a useful analytical method to demonstrate directly such phase transitions and change in the mixing behavior of the cellular lipid fractions in connection with mutation. A correlation between the alteration of the lipid composition in the plasma membrane of strains of S. cerevisiae S 288 and its mutant VY 1160 is discussed. PMID- 7473063 TI - The fix Escherichia coli region contains four genes related to carnitine metabolism. AB - Anaerobic carnitine metabolism in Escherichia coli was recently shown to involve six genes organized in the cai operon and located at the first minute on the chromosome. The DNA sequence lying at the 5' end of the cai locus was further investigated. It contains four open reading frames organized as an operon. In vivo overexpression of this DNA region revealed four polypeptides with apparent molecular masses of 27, 33, 45 and 6 kDa. These proteins displayed significant amino acid sequence homologies with polypeptides encoded by the fixABCX operons from Azorhizobium caulinodans and Rhizobium meliloti. The four ORFs were thus named fixABCX. The first two gene products were also found to share a high degree of sequence similarity with the subunits beta and alpha, respectively, of mammalian electron transfer flavoproteins, suggesting a role for these proteins in a redox reaction. A singly polycistronic 5 kb mRNA transcript was detected in Northern blots under anaerobic conditions in the presence of DL-carnitine. Expression of a fixA-lacZ transcriptional fusion was induced by L(-)-carnitine and crotonobetaine but not by D(+)-carnitine, gamma-butyrobetaine, glycinebetaine and choline as found previously for the carnitine pathway. Similarly, the fix operon was repressed by glucose and nitrate. Moreover, expression of the fix operon was induced by the global regulatory proteins CRP and FNR and repressed by the histone-like protein H-NS. All these regulatory proteins have been shown also to control expression of carnitine enzymes. Results from Northern blots and lacZ fusion studies indicate a common regulation of expression of fix and cai operons, which implies a physiological linkage between these two loci. PMID- 7473064 TI - The properties of the multicopy suppressor of the ogd1 mutation in yeast. AB - The 8.1 kb chromosomal fragment partially suppressing the ogd1 mutation in Saccharomyces cerevisiae has been cloned. The molecular analysis revealed that its suppressor gene codes for a natural glutamine tRNA(CAG) and maps on chromosome XIII in the upstream region of the URA10 gene. The multicopy plasmids containing this tRNA gene also suppressed the standard trp1-1 amber mutation and conferred the sensitivity of yeast cells to paromomycin and increased temperature. PMID- 7473065 TI - Proteolytic activity of Aeromonas caviae. AB - Aeromonas strains produce a variety of virulence factors including proteases. Studies on the kinetics of growth of Aeromonas caviae NRRL B-966 and its proteases suggest that the proteolytic activities are produced throughout the growth phase, with peak level occurring at stationary phase. A. caviae synthesize both intracellular and extracellular proteases with the latter account for major portion of the total activity. Optimum pH for the A. caviae proteolytic activity is at 7.0. A. caviae produces a thermoresistant protease, whose activity is dependent on Mg++ and Ca++ ions. Inhibition of proteolytic activity by phenyl methyl sulfonyl fluoride suggest the presence of a serine protease in A. caviae. Nitrogenous compounds enhance the proteolytic activity while carbohydrates tested in this study inhibit the activity. PMID- 7473066 TI - Immunochemical features of a macromolecule of Treponema denticola. AB - In this study the extraction and the immunochemical features of a lipopolysaccharide-like (LPSL) macromolecule of T. denticola strains 35405, 35404, 33521 and 11 were investigated. The yield of LPSL molecule ranged between 0.5-0.9% of the cell dry weight, it possessed Limulus amebocyte lysate clotting activity, and it contained glucosamine, phosphate, heptose, glucose, small amounts of KDO, myristic and beta hydroxy myristic acid. Sera obtained from healthy individuals (ADA type I) periodontitis, from 3-8 month old infants, or the mouse monoclonal antibody, diluted 1:2, against T. pallidum did not react with the LPSL antigens of T. denticola strains 35405, 35404, 33521, and 11. Sera from patients with ADA type III-IV periodontitis were reactive with two 8-14 kDa bands even at serum dilutions of 1:2000. Sera from patients with ADA type II periodontitis showed good antibody response to the 8-14 kDa band at a dilution of 1:50, but were weekly reactive, or nonreactive at serum dilutions of 1:200. This study indicates that extraction of a lipopolysaccharide-like macromolecule is feasible from the assay spirochetes, and this macromolecule may be used as an antigen for the diagnosis of ADA types II-IV periodontitis. PMID- 7473067 TI - Toxicity study of europium chloride in rats. AB - EuCl3.6H2O was administered by gavage for 28 consecutive days to groups of 10 male and 10 female rats (Slc:Wistar strain) at doses of 0, 40, 200, or 1000 mg/kg/day. Additional groups of male and female rats receiving the 0 and 1000 mg/kg doses were used to assess recovery after 14 days subsequent to cessation of compound administration. Body weights and food consumption were measured, and hematological, clinical biochemistry, and histopathological examination were performed. The concentrations of europium (Eu) and of essential elements in organs were determined by ICP-MS or ICP-AES. In the rats of each sex dosed at 200 and 1000 mg/kg, the body weight gain significantly decreased because of reduction in food consumption. Hyperkeratosis of the forestomach and eosinocyte infiltration of the stomach submucosa were found in both sexes receiving the 1000 mg/kg dose group, suggesting an irritation effect by EuCl3.6H2O. The Eu levels increased dose dependently in the liver, kidneys, spleen, and femurs, and the accumulated volume of Eu in these organs was estimated to be about 1/100,000 of the total dosed amount. The administration of EuCl3.6H2O increased the serum iron concentration in males and the serum total iron binding capacity in each sex and decreased cholinesterase activity in females in the 1000 mg/kg dosed group. Iron concentrations in the spleen and strontium concentrations in the femurs of rats of both sexes dosed at 1000 mg/kg were significantly decreased. We concluded that the no-observed-effect level is 200 mg/kg/day. Our investigation demonstrated that elemental analyses of organs is a useful approach to toxicological study. PMID- 7473068 TI - A comparison of the concentrations of certain chlorinated hydrocarbons and polychlorinated biphenyls in bone marrow and fat tissue of children and their concentrations in breast milk. AB - Chlorinated hydrocarbon (CHC) and polychlorinated biphenyl (PCB) concentrations in the bone marrow of 57 children were compared with the concentrations in adipose tissue of 50 children and the concentrations in breast milk in the Federal Republic of Germany from 1984 to 1991. The concentrations of hexachlorobenzene (HCB), the dichlorodiphenyltrichloroethane (DDT)-metabolites, and polychlorinated biphenyl (PCB) congeners no. 138 and no. 153 were increased threefold, while the concentrations of several hexachloro-cyclohexane (HCH) isomers and PCB congener no. 180 were only increased twofold. Because breast feeding is the primary source of CHC and PCB in toddlers and infants we also compared the concentrations in bone marrow of children with the concentrations in breast milk and found approximately fourfold higher concentrations for the most highly chlorinated PCB congener no. 180, but only threefold higher concentration for PCB 138 and 153 and the DDT-metabolites. The concentrations of beta-HCH and HCB were only slightly higher in bone marrow. PMID- 7473069 TI - Histopathological effects of acute exposure to chlorine gas on Sprague-Dawley rat lungs. AB - It is now recognized that acute inhalational exposure to irritant gases such as chlorine can cause an asthma-like abnormality known as reactive airways dysfunction syndrome (RADS). The aim of this study was to evaluate the effects of exposure to various levels of chlorine on airway mucosa and lung parenchyma in an attempt to develop an animal model of this syndrome. Seventy-four Sprague-Dawley rats were exposed to air (controls) or to 50, 100, 200, 500, and 1500 ppm of chlorine for 2 to 10 min. Histological assessment was performed at 1, 3, 6, 12, 24, and 72 h after exposure. The results show that exposure to low concentrations (500 ppm) did not induce significant histological changes. Exposure to 1500 ppm for 2 min induced perivascular edema and the appearance of focal mild inflammation, whereas exposure to the same concentration for 10 min caused profound histological changes, including (1) 1 h: airspace and interstitial edema associated with bronchial epithelial sloughing; (2) 6 to 24 h: decreased edema and the appearance of mucosal polymorphonuclear leukocytes, maximal at 12 h; (3) 72 h: epithelial regeneration, manifested by hyperplasia and goblet cell metaplasia. We conclude that acute exposure to chlorine at a concentration of 1500 ppm for 10 min induces significant airway mucosal abnormalities that vary over a short period of time. Whether these abnormalities are related to the subsequent development of RADS awaits further experiments. PMID- 7473070 TI - Overexpression of c-erbB-2 and p21 oncoproteins in human radiation-induced skin ulcers. AB - We studied the overexpression of c-erbB-2 and p21 oncoproteins in human radiation induced skin ulcers using immunohistochemistry. We found that the positive rate of overexpression of c-erbB-2 and p21 oncoproteins was 92.0 and 92.9%, respectively. The overexpression of c-erbB-2 oncoprotein was observed mainly in the cell membrane of squamous epithelial cells and in the cytoplasm of fibroblasts, endothelial cells, leiomyocytes in the media, and in fibrocytes of the adventitia of mesenchymal arterioles. The location of the p21 oncoprotein overexpression was mostly similar to that of c-erbB-2 with stronger staining in the cytoplasm of squamous epithelial cells and weaker staining in mesenchymal arteriolar walls. The overexpression of c-erbB-2 and p21 oncoproteins may be corresponding to the cancer transformation and poor healing of radiation-induced skin ulcers. PMID- 7473071 TI - Abnormal location of p16 protein and overexpression of p53 protein in human radiation-induced skin cancer. AB - To evaluate the role of p16 multiple tumor suppressor (MTS1) and p53 protein in human radiation-induced skin cancer, we examined the expression of p16 and p53 in eight cases using immunohistochemistry. An abnormal location of p16 expression in the cytoplasm was found in seven of eight cases, but the expression of p53 in nuclei was noted in only three of eight cases. These findings suggest that the inactivation of p16 might be related to its binding with another unknown oncoprotein in the cytoplasm and thus lead to carcinogenesis. PMID- 7473072 TI - Molecular biology studies on mesothelioma tumor samples: preliminary data on H ras, p21, and SV40. AB - The ras gene is one of the oncogenes most commonly detected in human cancers; this protooncogene is converted to active oncogene by point mutations occurring at either codon 12, 13, or 61. SV40 is a DNA-transforming simian virus, a 105 bp sequence of which has been shown recently to be present in a significant fraction of human mesothelioma cells. Eleven human malignant mesotheliomas were examined for H-ras gene mutations at codon 12, 13, and 61 and for the presence of SV40 like sequences. DNA prepared from formalin-fixed and paraffin-embedded tissue was amplified by means of PCR and analyzed using designed restriction fragment length polymorphism. No mutation with respect to H-ras was found in any tumor sample, but the majority of mesothelioma cells contained SV40-like sequences. PMID- 7473073 TI - Benzo[a]pyrene-induced respiratory tract cancer in hamsters fed a diet rich in beta-carotene. A histomorphological study. AB - The effect of a high dietary level of beta-carotene on the formation of preneoplastic and neoplastic respiratory tract lesions was studied in hamsters intratracheally treated with benzo[a]pyrene (B[a]P) attached to ferric oxide (Fe2O3) and suspended in saline. In addition to conventional histopathological examinations, the expression of cytokeratins and the glutathione S-transferase isoenzyme Pi (GST-Pi) was determined in tracheal epithelium using immunocytochemical techniques. B[a]P treatment increased the expression of cytokeratins in tracheal mucous and ciliated epithelial cells as detected by antibody RCK102 (cytokeratins 5 and 8), which normally recognizes basal cells only. The expression of cytokeratins in mucous and ciliated cells as detected by antibody RGE53 (cytokeratin 18) was decreased by B[a]P treatment. Furthermore, the expression of the cytokeratin detected by antibody RKSE60 (cytokeratin 10), characteristic of metaplastic squamous cells, and the expression of the GST-Pi, characteristic of metaplastic changes, was increased in trachael epithelium of hamsters treated with B[a]P. There was no evidence for dietary beta-carotene affecting the expression of cytokeratins or GST-Pi. The incidence of preneoplastic changes and tumors of the respiratory tract was not reduced by dietary beta-carotene. On the contrary, the tumor response of the respiratory epithelium was almost twice as high in hamsters fed the high beta-carotene diet than in hamsters on the low beta-carotene diet. However, this difference was not statistically significant (P = 0.15); hence, the present study did not produce evidence for a clear effect of beta-carotene on B[a]P-induced respiratory tract cancer in hamsters. PMID- 7473074 TI - Studies on hepatic xenobiotic-metabolizing enzymes in rats treated with insecticide deltamethrin. AB - The effects of the synthetic dibromo-pyrethroid insecticide deltamethrin on some hepatic phase I and II enzyme activities were studied in rat liver. The animals were treated with daily doses of 5 and 10 mg/kg of both pure insecticide or its commercial formulation (Decis), administered i.p. in corn oil for 7 days. The following enzyme activities were studied: NADPH-cytochrome-P450 reductase, aryl hydrocarbon hydroxylase, aminopyrine N-demethylase, glutamyl cysteine synthetase, glutathione S-transferase, glutathione peroxidase, peroxisomal acyl-CoA oxidase, catalase, and urate oxidase. Both deltamethrin and its commercial formulation were effective in modifying the activities of several of these hepatic xenobiotic metabolizing enzymes. However, some differences in enzyme modifications were found between treatment with pure or commercial deltamethrin, the latter being more active. This effect could be ascribed to additives, solvents, and chemical intermediates present in the Decis formulation. These results suggest that exposure to this deltamethrin commercial formulation could be more dangerous than exposure to deltamethrin alone, both in terms of its hepatotoxicity and/or alterations in the hepatic biotransformation of other occupational/environmental xenobiotics. PMID- 7473075 TI - Predictors of lead stores in male veterans. AB - Thirty-two male veterans participated in a study to determine cumulative lead exposure in an urban population. Subjects were chosen on the basis of blood pressure status in order to attempt to compare lead exposure between patients with and without hypertension. Patients currently enrolled in hypertension clinic and on treatment were recruited and matched with controls for age, race, and socioeconomic status. Each subject underwent provocative chelation via slow intravenous infusion of CaNa2 EDTA and 6-h urinary lead measurement and completed an interviewer-administered questionnaire. Twenty blacks and 12 whites participated, with a median age of 52 years (range: 27 to 72). Urinary lead excretion ranged from below detection limits to frankly toxic levels in an individual with heavy moonshine ingestion. Lead levels were higher than reported in other non-workplace populations. The distribution of lead values was skewed, as expected, with a median excretion of 75 mcg lead/6 h (corresponding to a median 24 degrees post-chelation urinary lead excretion of 286 mcg) and modal values between 50 and 75 micrograms lead. Levels of 95 mcg lead/6 h (corresponding to 24 degrees levels of 333 mcg lead) and above were considered "high" (N = 11) and the remainder were "low" (N = 21). Among those able to recall various characteristics of their first childhood dwellings, the presence of flaking paint in a multiple family dwelling was strongly associated with "high" lead excretion (X2 = 9.32, p = 0.009). Hypertensives excreted slightly more lead than nonhypertensives, although the difference was not statistically significant in this small sample. Lead excretion was not associated with current (treated) blood pressure determinations among hypertensives. However, lead excretion was associated with systolic pressure as recorded on entry to the hypertension clinic (N = 21, R2 = 0.24, p = 0.03). PMID- 7473076 TI - The relationship between receptor-effector unit heterogeneity and the shape of the concentration-effect profile: pharmacodynamic implications. AB - The apparent concentration-effect relationship is the ensemble of many effector units (such as individual cells or channels) that do not always exhibit a uniform stimulus-effect relationship. This concept is substantiated by many observations of heterogeneity in receptor-effector populations including hormone secreting cells, response to hormonal stimuli, activity pattern of second messengers, stimulus-evoked synaptic currents, and single ion channels. The relationship between drug concentration and magnitude of pharmacologic response is commonly described by the sigmoidal Emax model which was derived from the Hill equation. The sigmoidicity factor (N) in this model is assumed to be a pure mathematical parameter without physiological connotations. This work demonstrates that the numerical value of N (measured empirically) is the product of two factors: (i) the degree of heterogeneity of the effector subunits, i.e., the elemental component that upon drug stimulus contributes its pharmacological effect independently and does not interact with other subunits (it could range from a single receptor up to a whole tissue), and (ii) value of N*--the shape factor of the subunits' concentration-effect relationship. A special case of this approach occurs when N* > 5, which is an on-off case. Here N is determined by the distribution (density equation) of the subunit values. In case of heterogeneity of the microparameters of the effector subunits the apparent N will always have a lower value than N*. According to this theory it can be concluded that without knowledge of the distribution of the microparameters no mechanistic interpretation can be deduced from the apparent N value. If in the future N* can be determined by theoretical or experimental methods, the distribution function relating N* to N can be calculated. The relevance of this theory is increased in view of the progress being made in advanced research techniques which may enable us to determine the concentration-effect relationship at the level of the individual effector unit. PMID- 7473077 TI - Pharmacodynamic model for joint exogenous and endogenous corticosteroid suppression of lymphocyte trafficking. AB - The circadian pattern of the immune system correlates with that of circulating T helper cells and inversely with cortisol concentrations. Corticosteroids, both endogenous and exogenous, cause lymphocyte dimunition in blood by retention of cells in the lymphatic circulation. A physiologic pharmacodynamic model was developed to describe changes in circulating lymphocytes as a function of both endogenous cortisol and methylprednisolone concentrations. The model was applied to T-helper and T-suppressor cell data collected from six asthmatic men during baseline, after single-dose, and after 6 days of 20 mg daily methylprednisolone. The model described all phases of the study well. Baseline circadian rhythm of lymphocytes was related to cortisol concentrations. Multiple-dosing of methylprednisolone caused apparent tolerance and decreased the sensitivity of lymphocytes to corticosteroids by 116% and markedly reduced endogenous cortisol concentrations. A 60% increase in circulating T-helper cells was observed which could be accounted for by dual changes in receptor sensitivity and endogenous cortisol. PMID- 7473078 TI - Pharmacokinetics of bupivacaine enantiomers in sheep: influence of dosage regimen and study design. AB - Bupivacaine is used as a racemate. In previous studies the mean total body clearance of R(+)-bupivacaine was found to be greater than S(-)-bupivacaine by 65% after iv bolus dose of separate enantiomers and by 20% after iv infusion to steady state of racemate. The present studies were performed to determine whether different study designs using different iv dosage regimens could influence the pharmacokinetic parameters determined for either bupivacaine enantiomer. rac Bupivacaine.HCl was administered iv to 6 adult Merino ewes by bolus, brief infusion, and prolonged infusion. Arterial blood concentrations of R(+)- and S(-) bupivacaine were measured by enantioselective HPLC. These regimens consistently produced lower arterial blood concentrations of R(+)-bupivacaine than S(-) bupivacaine due to R(+)-bupivacaine having a greater initial dilution volume by 16 (95% CI = 3-29)%, volume of distribution at steady state equilibrium by 32 (95% CI = 17-32)% and mean total body clearance by 28 (95% CI = 21-35)%. The slow half-life of R(+)-bupivacaine, however, was found to be 15 (95% CI = 0-31)% longer than that of S(-)-bupivacaine. The difference between enantiomers in mean total body clearance thus was similar to the previous study based upon infusion to steady state of rac-bupivacaine. Differences in pharmacokinetics attributable to the dosage regimen consisted of a greater mean total body clearance for R(+) bupivacaine along with a smaller terminal half life with the bolus regimen and a longer half-life of S(-)-bupivacaine after prolonged infusion. Differences in pharmacokinetics between the bupivacaine enantiomers occurred consistently in both distribution and clearance but the magnitude of the effect was less than 50% in each case. Systematic differences in pharmacokinetics associated with the dosage regimen were found mainly in terminal half-life. Dosage regimen, thus, was found to influence the pharmacokinetic results found experimentally and is therefore a significant variable in its own right. PMID- 7473079 TI - Pharmacodynamic analysis of hematologic profiles. AB - We discuss the analysis of the myelosuppressive effects of chemotherapy. Such analyses examine hematologic data that arise by monitoring patients after treatment with high doses of chemotherapy. We propose a flexible approach for modeling such information and, using data collected as part of a Phase I study of an anticancer agent, show some interesting aspects of the data that become available after fitting models this way. PMID- 7473080 TI - Comparison of some control strategies for three-compartment PK/PD models. AB - In drug therapy, effective dosage strategies are needed to maintain target drug effects. The relationship between drug dose and drug effect is often described by pharmacokinetic/pharmacodynamic (PK/PD) models where typically the PK model has a multicompartment form and the PD model is the sigmoidal Emax model. The parameters in the PK/PD model are generally unknown in the individual patient, although prior knowledge may be available and can be updated after measurements of drug effect are taken during the therapy. This fact, together with the complexity of the PK/PD model, makes the control problem complex. This paper investigates several control strategies in the framework of a three-compartment PK model plus an effect site with a PD model. Using computer simulations under different assumptions, we show that a MAP (maximum a posteriori) Bayesian type of strategy is effective, nevertheless in high-risk situations a stochastic control strategy hedging against estimation errors provides better performance at computational cost. PMID- 7473081 TI - Measuring switchability and prescribability: when is average bioequivalence sufficient? AB - Recent work, beginning with that of Anderson and Hauck in 1990, has led to a general acceptance of the need to ensure switchability in bioequivalence testing for approval of generic drugs. In other applications of bioequivalence testing, prescribability may be sufficient. However, there is less acceptance of the need to change statistical procedures and study designs from those currently used to assess the current criterion of average bioequivalence. We propose easily interpreted measures of switchability and prescribability. These measures provide bases for assessing conditions under which average bioequivalence is not sufficient to ensure switchability and prescribability, and hence for which a procedure for individual or population bioequivalence is required. The required conditions are sufficiently tight that they cannot be presumed to hold. Thus, there are reasonable conditions for which current practice is not sufficient. An outcome of this development is a connection between two current approaches for assessing individual bioequivalence. PMID- 7473082 TI - Simple approximate formulas for calculating the time to clear drug and the time to accumulate drug when the plasma disposition curve of the drug is multiexponential. PMID- 7473083 TI - 1995 William J. Stickel Gold Award. High strain rate tissue deformation. A theory on the mechanical etiology of diabetic foot ulcerations. AB - Foot ulcerations are one of the most common and dangerous complications associated with chronic diabetes mellitus. Many studies have focused on neuropathy, in conjunction with elevated ground reactive forces, as the principal cause of these ulcerations. The authors discuss the mechanical cause of diabetic ulcerations at the cellular level. It is hypothesized that increased rate of tissue deformation associated with foot slap secondary to progressive motor neuropathy is the actual culprit, and not the magnitude of local pressure applied. The authors present a cellular model that shows that high rates of tissue deformation may result in elevated intracellular calcium concentrations, which may lead to cellular death, while comparable loads gradually applied do not. Furthermore, there is no significant difference in the response observed at 5 psi and 10 psi. Based on these findings, it is hypothesized that techniques such as ankle foot orthoses, which control the velocity of foot strike, may be useful in treating diabetic foot ulcerations. PMID- 7473084 TI - 1995 William J. Stickel Silver Award. Structural analysis of absorbable pin and screw fixation in first metatarsal osteotomies. AB - The structural characteristics of 4.0-mm stainless steel screws compared with 4.0 mm poly-L-lactic acid absorbable screws and 2.0-mm stainless steel Steinmann pins compared with 2.0-mm poly-L-lactic acid absorbable pins in oblique closing base wedge osteotomies of the first metatarsal were evaluated. The authors performed oblique closing base wedge using an osteotomy guide in six matched pairs of fresh frozen first metatarsal bones. Fixation was achieved with either a 4.0-mm stainless steel screw or poly-L-lactic acid absorbable screw. An additional five pairs of matched specimens were used to compare 2.0-mm stainless steel and poly-L lactic acid absorbable pins with the same approach. Specimens were loaded to failure with the Bionix Material Testing System at a constant rate of 0.166 mm/sec. A Student's t-test for paired samples was used with a 95% confidence interval to measure differences in ultimate load, ultimate displacement, and structural stiffness. There was not a significant difference in any of the parameters evaluated in the 4.0-mm poly-L-lactic acid absorbable versus stainless steel screw comparison (P > 0.05). Stainless steel 2.0-mm pins had significantly greater structural stiffness (P = 0.032) and less ultimate displacement (P = 0.021) than their poly-L-lactic acid absorbable counterparts. There was not a significant difference in ultimate load in poly-L-lactic acid absorbable and stainless steel pins (P = 0.59). PMID- 7473085 TI - 1995 William J. Stickel Bronze Award. Prevalence of mixed infections in the diabetic pedal wound. A retrospective review of 112 infections. AB - This retrospective study reviewed the culture results of 112 admissions to a multidisciplinary diabetic foot care team with a primary diagnosis of infected diabetic pedal ulceration. An average of 1.5 +/- 0.9 species per patient (P < 0.0001) were isolated. Eighty-nine percent of wounds cultured grew two or fewer organisms. Anaerobic species were isolated in only 5% of all cultures. Of these isolates, the distinction between anaerobic colonization and true anaerobic infection is made. Results suggest that aggressive early hospitalization, coupled with aggressive intraoperative debridement, may yield less microbiologically complex infections that may be controlled with less expensive narrow spectrum antibiotic therapy. Diagnosis of the infected pedal ulceration of a patient with diabetes is a clinical one. If this diagnosis is combined with appropriate surgical intervention, microbiologic correlation, and antimicrobial therapy, the result may be a less complex hospital course and improved outcome. PMID- 7473086 TI - Misdiagnosed recalcitrant heel pain associated with HLA-B27 antigen. PMID- 7473087 TI - Osteochondroma of the distal tibia. A case study. AB - Osteochondromas can undergo malignant transformation, and although the incidence is low, it is important to recognize symptoms that might suggest transformation. Complaints of increasing pain or a dramatic increase in the size of the lesion must not be ignored. If the physician is confident that the lesion is benign and chooses not to resect it, the patient should be monitored periodically with serial radiographs in order to identify any potential degenerative changes that might develop. Since osteochondromas are usually associated with the growth plate of a long bone, the potential for progressive growth exists. This growth takes place through the process of enchondral ossification until closure of the physis occurs. After physiologic fusion of the physis, remnants of the cartilaginous cap may remain, possibly explaining the occasional development of peripheral chondrosarcomas later in life. PMID- 7473088 TI - Podiatric manifestation of Munchausen's syndrome. PMID- 7473089 TI - Squamous cell carcinoma of the nail bed. PMID- 7473090 TI - Melanoma appearing on the foot of an immunocompromised female. PMID- 7473091 TI - Peripheral nerve tumors. Large neurofibroma of the foot. AB - A neurofibroma of the foot is an uncommon finding and may be associated with von Recklinghausen's disease. A case of a large, solitary neurofibroma is presented with no apparent systemic involvement on history and physical examination. The surgical approach and perioperative treatment for this lesion is presented, and a review of peripheral nerve tumors and neurofibromatosis. A tumor of this size poses many unforeseen problems: large dead space, wound dehiscence, and infection caused by hematoma formation. Preoperative magnetic resonance imaging to determine position, depth, and involvements of vital structures is invaluable. The lazy S incision allows for greater exposure with less skin tension from retraction, thus reducing the risk of wound dehiscence. Careful dissection prevents cutting or injuring of underlying atrophied tissue. Two years after surgery, the plantar muscle has returned to normal strength and size and only a slight amount of sensory deficit exists. His gait is normal, without limp, and with normal heel-to-toe ambulation. This patient's life-style has returned to normal with no alterations. PMID- 7473092 TI - Ochronosis and alkaptonuria. Case report and literature review. PMID- 7473093 TI - Paget's disease of bone presenting as chronic ankle pain. PMID- 7473094 TI - Pruritic urticarial papules and plaques of pregnancy. PMID- 7473095 TI - Lancefield group C streptococcal arthritis and osteomyelitis. PMID- 7473096 TI - Indications and timing of cataract surgery. PMID- 7473097 TI - Atonic pupil. PMID- 7473099 TI - Scleral tunnel incision. PMID- 7473098 TI - Preventing internal wound gape with the scleral tunnel incision. PMID- 7473100 TI - Temporal approach with patients unable to lie flat. PMID- 7473101 TI - Early lens exchange for power calculation error. PMID- 7473102 TI - Planned eccentric lens implantation. PMID- 7473103 TI - Topical anesthesia in patients with communication deficits. PMID- 7473104 TI - Consultation section. A 73-year-old male with uncomplicated phacoemulsification in his right eye. PMID- 7473105 TI - Phaco sweep. AB - We present a variation on a technique for down slope sculpting designed to enhance the safety of sculpting deeply in the central nucleus. The phaco tip moves laterally to remove very deep nuclear material, increasing visualization with less risk of occlusion and break through to the capsule. PMID- 7473106 TI - Repositioning a tilted ciliary-sulcus-fixated intraocular lens. PMID- 7473107 TI - Deep, topical, nerve-block anesthesia. AB - Retrobulbar anesthesia produces profound anesthesia but involves risks such as hemorrhage and ocular tissue damage. Simple topical anesthesia is safer but does not produce the same depth of anesthesia. I have developed a technique that places a lidocaine-soaked sponge deep in the conjunctival fornices. This deep, topical, "nerve-block" technique produces a level of anesthesia previously seen only with injection techniques. I present the results of using this technique in 81 cataract extraction and intraocular lens implantation procedures. Supplemental injection was used in only two patients. The technique has advantages over injection and topical methods of anesthesia and is applicable to a variety of surgical procedures. PMID- 7473108 TI - Sutureless scleral tunnel cataract surgery using topical and low dose perilimbal anesthesia. AB - Because of risks and complications inherent to retrobulbar anesthesia, alternative techniques such as peribulbar and topical approaches have been devised. These also have associated problems. We have developed a technique that combines a single, minimal volume, perilimbal injection with topical anesthesia. It provides sutureless scleral tunnel cataract surgery without the risks of retrobulbar or peribulbar anesthesia, maximizing patient comfort and allowing immediate return of vision postoperatively. PMID- 7473109 TI - Myopic keratomileusis in situ combined with VISX 20/20 photorefractive keratectomy. AB - We present the results of myopic keratomileusis in situ combined with photorefractive keratectomy (PRK) in five eyes with high myopia and low astigmatism. Follow-up ranged from 10 to 12 months. A hinge was used to perform subtotal lamellar keratectomy and the spherical component of myopia was corrected by three-zone PRK. The mean preoperative spheroequivalent was -21.00 diopters (D) and the postoperative spheroequivalent was -1.50 D. In four eyes the best corrected visual acuity was unchanged or improved; in one eye it decreased because of epithelial ingrowth in the lamellar interface. Persistent oblique longitudinal folds were noted in the corneal disc in one eye, and induced cylinder was found postoperatively in one eye. PMID- 7473110 TI - Correcting simple myopic astigmatism with the excimer laser. AB - For one year we followed 46 eyes that had photoastigmatic refractive keratectomy with an excimer laser to correct simple myopic astigmatism. Mean preoperative cylinder was -2.50 +/- 0.70 diopters (D); mean preoperative best corrected visual acuity was 20/25 +/- 1 Snellen line. Mean postoperative refraction one year after surgery was -0.5 +/- 0.2, and mean uncorrected visual acuity was 20/25 +/- 1 Snellen line. In all eyes, the final axis of the residual cylinder was +/- 5 degrees around the preoperative axis. Uncorrected visual acuity equaled best preoperative acuity in 24 eyes. Eight eyes gained one line of best corrected visual acuity, six gained more than one line, and eight lost one line. No patient lost more than one line of uncorrected visual acuity. No significant complications such as haze, halos, eccentric ablation, or hypercorrection were observed. PMID- 7473112 TI - Small-diameter corneal inlay in presbyopic or pseudophakic patients. AB - We report the results of using the small-diameter corneal inlay to create a bifocal cornea. The inlay was implanted in five eyes in January 1993. At 12 months postoperatively, uncorrected near vision had improved from J4 to better than J2 in four of the five. The results indicate that the corneal inlay improves near vision and is compatible within the cornea. PMID- 7473111 TI - Corneal subepithelial infiltrates following excimer laser photorefractive keratectomy. AB - Corneal subepithelial opacification associated with pain, photophobia, and injection has been reported in the first to third day following photorefractive keratectomy (PRK). Regardless of treatment, the resolution has generally left the patient with permanent corneal scarring and a one to two line reduction in visual acuity. We surveyed 50 PRK surgeons and received responses from 17 on 30 cases of this complication, which occurs in approximately one in 300 cases. The etiology is unknown. Cases were first reported when nonsteroidal anti-inflammatory drugs, with or without occlusive contact lens, were substituted for conventional bandage occlusion in the postoperative protocol for PRK. PMID- 7473113 TI - Cataract surgery in eyes with early cataracts. AB - This prospective study assessed visual results and subjective rehabilitation after cataract surgery in eyes with a preoperative acuity of 0.4 (20/40) or better. Fifty eyes (50 patients) were entered in the study. Corrected and uncorrected visual acuity, lens opacity, glare disability, and contrast sensitivity were measured before surgery and four months after surgery. Patient satisfaction was also measured. Best corrected visual acuity was 0.54 +/- 0.12 before surgery and 0.95 +/- 0.15 four months after surgery. Uncorrected visual acuity was 0.24 +/- 0.18 before and 0.58 +/- 0.27 after surgery. Impairment of corrected visual acuity from glare decreased from 0.12 +/- 0.09 before surgery to 0.04 +/- 0.08 four months after surgery. Contrast sensitivity at 6 cycles/degree increased from 2.71 +/- 0.94 units to 4.73 +/- 1.09 units. After surgery, 62% of patients judged their vision to be greatly improved, 32% judged it improved, 6% judged it unchanged. The patient's satisfaction was partially related to the improvement in uncorrected visual acuity. We conclude that cataract surgery can benefit eyes with limited visual impairment if the surgery is uncomplicated and IOL power is calculated correctly. PMID- 7473114 TI - Combined phacoemulsification, intraocular lens implantation, and trabeculectomy with a modified scleral tunnel and single-stitch closure. AB - In most combined phacoemulsification and filtration surgery, the standard tunnel incision is altered to create a scleral flap, usually a limbal-based scleral flap with radial suture closure. We describe a new technique that has the advantages of both scleral tunnel architecture and horizontal suture closure. It creates a scleral groove tangential to the limbus and a standard scleral tunnel. Phacoemulsification and intraocular lens implantation are performed under the scleral tunnel. A 10-0 nylon horizontal suture is positioned for scleral tunnel closure and looped aside. A central "T" relaxing incision is made at the midpoint of the scleral tunnel up to the posterior insertion of conjunctiva. A punch sclerectomy is performed. The 10-0 nylon horizontal suture is tied. The conjunctiva is closed and bleb elevation is confirmed via deepening through a corneal paracentesis incision. The results in an initial series of 24 consecutive eyes followed for a mean of seven months are presented: Mean visual acuity improved from 20/70 to 20/30 and mean intraocular pressure reduction was 5.0 +/- 2.4 mm Hg (standard deviation). By preserving standard scleral tunnel architecture and single horizontal suture closure, this technique permits rapid visual recovery and accelerated return to full activity. It also permits postoperative titration of aqueous outflow by laser suture lysis, if indicated. Against-the-rule astigmatic drift associated with limbal-based scleral flaps and antimetabolite therapy may be minimized. PMID- 7473115 TI - Visual outcome following traumatic wound dehiscence after cataract surgery. AB - We reviewed the charts of 21 patients who had cataract surgery and subsequent ocular trauma with wound dehiscence. Eighteen patients were re-examined for this study; the other three were lost to follow-up. A complete examination of both eyes was performed 3 to 84 months after the trauma occurred. Patients were assigned to one of two groups: Group A, minor trauma (n = 14), or Group B, major trauma (n = 7). In Group A, visual acuity was 20/40 or better in four patients, 20/50 to 20/80 in five patients, and 20/300 or worse in three patients. In Group B, visual acuity was 20/60 or better in three patients and 20/300 or worse in three patients. Poor visual acuity was associated with endophthalmitis and bullous keratopathy in Group A and globe rupture or recurrent retinal detachment in Group B. In 14 of the 18 patients, final visual acuity in the traumatized eye was similar to that in the fellow eye, which had the same ocular pathology. This study suggests that pre-existing ocular disease is an important factor in final visual outcome in patients with traumatic wound dehiscence after cataract surgery. PMID- 7473116 TI - Anterior chamber fluid contamination after uncomplicated phacoemulsification. AB - This study determined the rate of anterior chamber aerobic bacterial contamination in 103 eyes that had uncomplicated posterior chamber phacoemulsification. Anterior chamber fluid was aspirated on completion of surgery. Cultures of sterile balanced salt solution served as a control. Of 103 anterior chamber cultures, 4 (3.9%) showed contamination; however, 3 of 103 control cultures (2.9%) also grew micro-organisms. These findings suggest a very low rate of aerobic bacterial contamination after uncomplicated phacoemulsification and underscore the importance of control cultures when determining contamination rates. PMID- 7473117 TI - Clinical comparison of Provisc and Healon in cataract surgery. AB - This prospective, randomized, multicenter clinical trial compared the safety and efficacy of the Provisc and Healon viscoelastics. Sixty-one eyes of 61 patients had an extracapsular cataract extraction with implantation of a posterior chamber intraocular lens with the aid of Provisc (n = 32) or Healon (n = 29). Mean changes in preoperative versus postoperative corneal thickness and intraocular pressure and the incidence and magnitude of postoperative corneal edema and iritis were not significantly different between the two groups. Observed complications were considered consequences of the surgery and unrelated to viscoelastic use. No adverse medical events occurred. These results indicate Provisc and Healon are clinically equivalent in terms of safety and efficacy when used as surgical aids. PMID- 7473118 TI - Postoperative complications of intraocular lens implantation in patients with Fuchs' heterochromic cyclitis. AB - We studied 29 eyes of 24 patients with Fuchs' heterochromic cyclitis who had extracapsular cataract extraction and posterior chamber intraocular lens implantation. Follow-up ranged from one to four years (mean two years). Intraoperative complications included mild hyphema (n = 4). Early postoperative complications included uveitis (n = 9), glaucoma (n = 7), pigment deposits on the lens surface (n = 8), vitreous opacities (n = 8), and cystoid macular edema (n = 1). Late postoperative complications included posterior capsule opacification (n = 6), recurrent uveitis (n = 4), and persistent glaucoma (n = 3). Best corrected visual acuity of 20/40 or better was achieved in 24 eyes (82.8%). PMID- 7473119 TI - Theoretical analysis of image defocus with intraocular lens decentration. AB - We used a theoretical eye model to evaluate the effect on peripheral images when planoconvex, biconvex, and meniscus intraocular lenses (IOLs) decenter 1 mm or 2 mm upward or downward from the pupillary center. A light source was projected from an angle of 0, 20, 28, or 40 degrees downward to a retina with a radius of curvature of 11 mm. When the IOL was centered, the light focused at a single point on the retina. When the IOL was decentered, the light became more dispersed as the incident angle of the light increased. The most dispersion was noted with the meniscus IOL; the least with the biconvex IOL. Astigmatism and aberration, which IOL decentration increases, were thought to cause the image defocus. These results suggest that IOL decentration may affect the sharpness of peripheral images. PMID- 7473120 TI - Effect of age and astigmatism on the AMO Array multifocal intraocular lens. AB - Multifocal intraocular lenses (MIOLs) have been designed to provide better uncorrected near visual acuity than monofocal IOLs. However, the increase in depth of focus is combined with a significant decrease in contrast sensitivity and glare. We implanted a new zonal-progressive MIOL (AMO Array) in 31 eyes of 25 patients whose mean age was 60.8 +/- 13.5 years. At 12.1 +/- 2.4 months postoperatively, the uncorrected visual acuity averaged 0.56 +/- 0.09 (20/32) for distance and 0.54 +/- 0.12 (20/40) for near focus. Seventy-five percent of patients achieved a best corrected distance visual acuity of > or = 0.9 (20/22). With distance correction only, mean near visual acuity was 0.72 +/- 0.17 (20/25). Best corrected near acuity reached 0.89 +/- 0.11 (20/22), requiring an additional near add of +1.0 to +2.0 diopters (D). We observed an age-dependent difference in the depth of focus. In younger patients (46 +/- 6 years), the pseudoaccommodation range was 6.25 D; acuity attained was > or = 0.4 (20/50). In older patients (71 +/- 7 years), visual acuity at the near focus was substantially reduced (two lines). Regan contrast sensitivity was lower for MIOL patients at all contrast levels, but the difference was statistically significant only at very low contrast (11%). Contrast sensitivity and glare were dependent on the patient's age but did not change with the amount of postoperative astigmatism. Younger patients were very satisfied with the results of MIOL implantation, and the small loss of contrast sensitivity was counterbalanced by the advantage of improved depth of focus and uncorrected near visual acuity. PMID- 7473121 TI - Complications of silicone disc intraocular lenses. AB - Through a 4.5 mm corneoscleral incision, we implanted one of two types of foldable, silicone disc, posterior chamber intraocular lenses (IOLs) in the bag in 35 eyes of 32 patients after phacoemulsification and capsulorhexis. Five lenses were explanted in the early postoperative period because of complications. We followed 24 eyes for an average of 38 months. At the end of follow-up, 5 of the lenses had decentered by 1 mm; 8 (33%) had developed clinically significant posterior capsule opacification (PCO). We believe the IOLs' disc shape may not allow firm fixation, resulting in opacification. On average, there were no significant differences in long-term results between the two silicone disc lens types. PMID- 7473122 TI - Transscleral fixation of posterior chamber intraocular lenses. AB - In a retrospective study of 75 consecutive eyes having transscleral suturing of a posterior chamber intraocular lens (IOL) (mean follow-up five months), 57% had improved best corrected visual acuity; 67% of those obtained a visual acuity of 20/40 or better. Thirty-seven eyes had an IOL exchange without penetrating keratoplasty (PKP) at the time of implantation, 15 had simultaneous PKP, and 23 had secondary lens implantation without IOL exchange or a corneal transplant. Transscleral fixation is an effective means of implanting a posterior chamber lens in eyes without capsular support. It allows secure two-point lens fixation with minimal risk of IOL tilting or suture erosion through the conjunctiva or sclera. PMID- 7473123 TI - Effect of indomethacin-coated posterior chamber intraocular lenses on postoperative inflammation and posterior capsule opacification. AB - We implanted indomethacin-coated intraocular lenses (IOLs) in rabbit eyes to ascertain how the sustained release of the drug affects postoperative inflammation and posterior capsule opacification (PCO). A preoperative resolution test showed that 1% indomethacin-coated IOLs placed in balanced salt solution released a mean 14.0 +/- 0.6 micrograms of the drug within 24 hours. Postoperatively, aqueous flare intensity in eyes with 0.1% indomethacin-coated IOLs and in eyes with 1.0% indomethacin-coated IOLs was significantly less at day 2 and at days 2 and 3, respectively, than in control eyes with uncoated lenses. In another group of eyes, histopathological examination showed that those with 1.0% indomethacin-coated lenses had significantly less PCO than the contralateral control eyes with uncoated IOLs. PMID- 7473124 TI - Comparison of the postoperative inflammatory response in the normal eye with heparin-surface-modified and poly(methyl methacrylate) intraocular lenses. AB - Evidence that poly(methyl methacrylate) (PMMA) is not inert in the eye has led to the introduction of intraocular lens (IOL) surface modification. In this prospective, double-blind, randomized study, we evaluated the effects of heparin surface modification (HSM) on anterior segment inflammation for one year after endocapsular cataract surgery. Fifty-four eyes were randomized to receive an HSM IOL (29 eyes) or a PMMA IOL (25 eyes) and were assessed postoperatively by corneal endothelial photography, laser flare and cell measurements, fluorophotometry, and IOL surface specular microscopy. Corneal endothelial loss, mean aqueous flare and cells, and fluorophotometry did not differ significantly between the groups, although fewer eyes in the HSM group had high flare values on the first postoperative day. The number of giant cells was significantly less in the HSM group for up to one year after surgery. Although there was no statistical difference in overall visual outcome, more patients in the HSM group (17%, 5/29) achieved a postoperative visual acuity of 20/15 than in the PMMA group (0). These results demonstrate that HSM enhances IOL biocompatibility and these effects are detectable in the low-risk, normal eye. PMID- 7473125 TI - Effects of the picosecond neodymium:YLF laser on poly(methyl methacrylate) intraocular lenses during experimental posterior capsulotomy. AB - We evaluated the effect of the neodymium:YLF picosecond laser on intraocular lenses (IOLs) during experimental posterior capsulotomy and established the minimum damage threshold for lathe-cut poly(methyl methacrylate) (PMMA) IOLs. Single-piece, biconvex, lathe-cut PMMA lenses were placed in a model eye chamber against a latex membrane with a central area of lens-capsule touch. Energy levels from 40 micro J to 360 microJ pulse (0.4 mJ to 3.6 mJ) with a 0.5 mm line and a spot pattern were tested using different focal offsets. Threshold damage level was determined under the scanning electron microscope after observing the minimal energy level that did not cause damage to or pits in the IOLs. Lenses were then analyzed for size and type of damage. The 0.5 mm line pattern at 160 microJ using 350 microns offset was safe and effective for posterior capsulotomy in vitro. spot pattern caused IOL damage in the form of small pits at much lower energy levels of 80 microJ using 400 microns offset than the line pattern. PMID- 7473126 TI - Anti-dopamine beta-hydroxylase immunotoxin-induced sympathectomy in adult rats. AB - Anti-dopamine beta-hydroxylase immunotoxin (DHIT) is an antibody-targeted noradrenergic lesioning tool comprised of a monoclonal antibody against the noradrenergic enzyme, dopamine beta-hydroxylase, conjugated to saporin, a ribosome-inactivating protein. Noradrenergic-neuron specificity and completeness and functionality of sympathectomy were assessed. Adult, male Sprague-Dawley rats were given 28.5, 85.7, 142 or 285 micrograms/kg DHIT i.v. Three days after injection, a 6% to 73% decrease in the neurons was found in the superior cervical ganglia of the animals. No loss of sensory, nodose and dorsal root ganglia, neurons was observed at the highest dose of DHIT. In contrast, the immunotoxin, 192-saporin (142 micrograms/kg), lesioned all three ganglia. To assess the sympathectomy, 2 wk after treatment (285 micrograms/kg), rats were anesthetized with urethane (1 g/kg) and cannulated in the femoral artery and vein. DHIT treated animals' basal systolic blood pressure and heart rate were significantly lower than controls. Basal plasma norepinephrine levels were 41% lower in DHIT treated animals than controls. Tyramine-stimulated release of norepinephrine in DHIT-treated rats was 27% of controls. Plasma epinephrine levels of DHIT animals were not reduced. DHIT-treated animals exhibited a 2-fold hypersensitivity to the alpha-adrenergic agonist phenylephrine. We conclude that DHIT selectively delivered saporin to noradrenergic neurons resulting in destruction of these neurons. Anti-dopamine beta-hydroxylase immunotoxin administration produces a rapid, irreversible sympathectomy. PMID- 7473127 TI - Interindividual variation in expression of P-glycoprotein in normal human liver and secondary hepatic neoplasms. AB - P-glycoprotein (Pgp), a drug transport protein, pumps many drugs out of hepatocytes. To begin to determine how variation in the level of human hepatic Pgp might influence individual differences in drug disposition, we have used Northern blot and immunochemical analysis to determine the variation in Pgp and in the mRNA for Pgp (MDR1) in liver from 41 individuals. These samples were divided into two groups, normal and perineoplastic (normal liver adjacent to secondary hepatic neoplasms). There was large variation in MDR1 mRNA and Pgp protein expression between all human liver samples. The average amount of Pgp was 2.5-fold greater in normal than in perineoplastic liver. Hepatic Pgp expression was associated with gender, with males expressing 2-fold higher amounts of Pgp than females. There was no correlation between expression of MDR1 and cytochrome P4501A1, but there was a trend toward Pgp and cytochrome P4503A proteins being inversely correlated, although it did not reach statistical significance. MDR1 expression was increased in three of four individuals who had previously received chemotherapy. Pgp expression appeared to be regulated developmentally as MDR1 mRNA was undetectable in six fetal livers, but Pgp was present as early as 1 month postnatally. The level of Pgp was then compared between nine paired samples consisting of seven secondary metastatic hepatic neoplasms, one primary heptocellular carcinoma, one hepatic adenoma and their adjacent normal perineoplastic liver. There was no consistent increase or decrease in Pgp expression in secondary hepatic neoplasms compared with paired perineoplastic liver.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7473128 TI - Amphetamine produces sensitized increases in locomotion and extracellular dopamine preferentially in the nucleus accumbens shell of rats administered repeated cocaine. AB - Alterations in dopamine transmission in the nucleus accumbens, which is composed of two anatomically distinct compartments termed the shell and core, contribute to the expression of cocaine-induced behavioral sensitization. To test potential presynaptic components of behavioral sensitization, the behavioral and neurochemical response to amphetamine administration in the accumbens shell and core was measured at early (days 1-3) and late (days 20-22) withdrawal in rats pretreated with systemic cocaine (15 mg/kg x 2 days, 30 mg/kg x 5 days) or saline. Behavioral sensitization was observed at late, but not early withdrawal when amphetamine was microinjected into the nucleus accumbens shell of cocaine pretreated rats. There were no significant differences between cocaine- and saline-pretreated animals when behavior was monitored after amphetamine injections into the core at either withdrawal period. After both withdrawal periods, the amphetamine-induced increase in extracellular dopamine was potentiated among cocaine-pretreated animals in the shell by the local administration of amphetamine (0.03, 0.3, 3.0 and 30 microM through the dialysis probe). In the core at early withdrawal there was tolerance to the amphetamine induced increase in extracellular dopamine in the cocaine group, whereas there was no difference between the repeated saline and cocaine groups at late withdrawal. In a second experiment designed to evaluate potential postsynaptic influences, the D1 partial agonist, SFK-38393 (0.01 or 0.1 microgram/side), was microinjected into the nucleus accumbens core or shell regions after behavioral sensitization to cocaine. Although there was a motor-stimulant effect of SKF 38393 at both withdrawal periods, there was no difference between rats pretreated with repeated cocaine or saline. Collectively, these results demonstrate that the augmentation in dopamine transmission in the nucleus accumbens that is associated with behavioral sensitization is more robust in the shell than the core. PMID- 7473129 TI - [3H]epibatidine labels nicotinic receptors in rat brain: an autoradiographic study. AB - Epibatidine, a frog skin alkaloid, is a nicotinic receptor agonist with potent analgesic activity. We used in vitro receptor autoradiographic techniques to map binding sites for [3H]epibatidine in rat brain, and those for another nicotinic agonist, [3H]cytisine. Both [3H]epibatidine and [3H]cytisine are excellent radioligands for autoradiography. [3H]Epibatidine in particular binds with very high affinity and extremely low nonspecific binding. In the present study, the distribution of binding was almost identical for both radioligands and closely resembled that of previously published autoradiographic studies with two other nicotinic ligands, [3H]acetylcholine and [3H]nicotine. Brain regions demonstrating the highest levels of [3H]epibatidine binding included the interpeduncular nucleus, medial habenular nucleus, fasciculus retroflexus, superficial gray layer of the superior colliculus and numerous thalamic nuclei, including the anteroventral, dorsal lateral geniculate and gelatinosus nuclei. Quantitative analysis revealed that [3H]epibatidine binding was greater than [3H]cytisine binding in several regions. In particular, [3H]epibatidine clearly labeled the optic nerve, optic chiasm and optic tract, whereas [3H]cytisine binding to these structures did not exceed background levels. Other regions in which [3H]epibatidine binding was greater than that of [3H]cytisine included the medial habenula, fasciculus retroflexus, olivary pretectal nucleus and superficial gray layer of the superior colliculus. The differences in labeling between these two nicotinic ligands may arise from receptor heterogeneity in brain nicotinic receptors and the ability of [3H]epibatidine to label more than one subtype of nicotine receptor. PMID- 7473130 TI - Quantification of steady state expression of mRNA for alpha-1 adrenergic receptor subtypes using reverse transcription and a competitive polymerase chain reaction. AB - Three distinct alpha-1 adrenergic receptor subtypes (alpha-1a, alpha-1b and alpha 1d) have been identified through molecular cloning. Expression of alpha-1 adrenergic receptor mRNA has been detected in several tissues, but previous studies with Northern blot analysis or RNase protection assays have provided only semiquantitative information. Furthermore, the mRNA distribution of these subtypes in many rat tissues is unknown. In the present study, we quantified alpha-1a, alpha-1b and alpha-1d adrenergic receptor mRNA in 19 adult rat tissues through the use of reverse transcription (RT) and a competitive polymerase chain reaction (PCR). This procedure used internal cRNA standards as competitive templates that contained sequences complementary to the primers for alpha-1a, alpha-1b and alpha-1d adrenergic receptors and that differed in molecular size from the native sequences. Total RNA was harvested from 19 freshly dissected organs of the rat. The PCR products were labeled by inclusion of alpha-32P-dCTP in the PCR. After electrophoresis, ratios of competing to native radio-labeled products were used to interpolate the amount of native RNA originally present in each sample. The results revealed that the relative abundance of alpha-1a adrenergic receptor mRNA among various tissues was as follows: vas deferens > colon > or = stomach > or = cerebral cortex > heart > or = small intestine > or = testis > prostate. The relative rank order of alpha-1b adrenergic receptor mRNA expression was heart > or = cerebral cortex > liver, whereas that of alpha-1d adrenergic receptor mRNA was vas deferens > cerebral cortex > or = aorta > adrenal gland.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7473131 TI - The immunosuppressant leflunomide inhibits lymphocyte proliferation by inhibiting pyrimidine biosynthesis. AB - Leflunomide is a novel immunosuppressive compound that is effective in the treatment of animal models of autoimmune disease and human rheumatoid arthritis. The mechanism of action is unknown. Here we show that leflunomide blocked 1) increases in nucleolar size and number, 2) upregulation of the nuclear protein antigens (PCNA and Ki-67), 3) increases in uridine incorporation and total RNA and DNA content, 4) cell cycle progression and 5) proliferation in mitogen stimulated rat spleen mononuclear cells and human peripheral blood mononuclear cells (HPBMC). Exogenous uridine reversed the leflunomide-dependent inhibition of the normal increase in total RNA and DNA content in mitogen-stimulated HPBMC and rat spleen cells. Uridine reversed the leflunomide-dependent inhibition of cell cycle progression in stimulated rat cell cultures. Either uridine or cytidine, which can be converted to uridine by cytidine deaminase, reversed the antiproliferative effect of leflunomide in HPBMC. Dihydroorotate accumulated in leflunomide-treated human T-lymphoblastoid cells, suggesting that the compound inhibited the fourth enzyme in the pyrimidine biosynthetic pathway, dihydroorotate dehydrogenase. The results support the hypothesis that the in vitro effects of leflunomide on T-lymphocytes are due to inhibition of de novo pyrimidine synthesis. PMID- 7473132 TI - Neurochemical and behavioral evidence that quipazine-ketanserin discrimination is mediated by serotonin2A receptor. AB - The purposes of this study were to determine: first, if animals could be trained to discriminate a serotonin (5-HT)2 receptor agonist from a 5-HT2 receptor antagonist; second, which 5-HT2 receptor subtype was mediating the cues; and third, the usefulness of this model for studying adaptive changes in the 5-HT2 receptor system. Rats were trained to discriminate quipazine (0.5 mg/kg) from ketanserin (1.0 mg/kg) on a variable interval-20 schedule of reinforcement. After acquisition, the quipazine and ketanserin dose-response curves were found to be orderly and reproducible. Additional 5-HT2 receptor agonists (2,5-dimethoxy-4 iodoamphetamine and MK 212) and antagonists (pizotifen, mianserin, pirenperone and MDL 100,907) were tested for generalization and found to substitute for the quipazine and ketanserin cues, respectively. In antagonist studies, MDL 100,907 potently blocked quipazine discrimination. Results of ex vivo binding studies designed to estimate occupancy of 5-HT2A and 5-HT2C receptors suggested that the training dose of ketanserin blocked only 5-HT2A receptors and not 5-HT2C receptors. The combined results from the substitution, antagonism and ex vivo receptor autoradiographic studies suggest that the discriminative stimuli of quipazine and ketanserin are mediated at least in part by the 5-HT2A receptor. Additional experiments were designed to study adaptive changes in the 5-HT2A receptor. A single large dose of quipazine produced a rebound ketanserin-like effect at 20 hr after administration; however, a single large dose of ketanserin (10 mg/kg) did not produce a rebound quipazine-like effect. PMID- 7473133 TI - Cyclopiazonic acid, inhibiting the endoplasmic reticulum calcium pump, reduces the canine colonic pacemaker frequency. AB - The slow wave frequency of the canine colon has previously been hypothesized to be paced by an intracellular biochemical clock. We investigated the relationship between the endoplasmic reticulum (ER) Ca++ and the periodicity of the biochemical clock. Cyclopiazonic acid, a specific inhibitor of the ER Ca++ pump, dose-dependently decreased the pacemaker frequency. Similarly, chelating cytosolic Ca++ with bis-(o-aminophenoxy)-ethane-N,N,N',N'-tetra-acetic acid (BAPTA) also decreased the pacemaker frequency. These observations suggest that delaying the Ca++ uptake into the ER decreases the pacemaker frequency. The pacemaker frequency was similarly decreased by neomycin [inhibiting inositol 1,4,5-triphosphate (IP3) synthesis] and by caffeine at concentrations higher than 5 mM (inhibiting the IP3-sensitive Ca++ channels in the ER membrane). Hence the IP3-sensitive Ca++ stores are involved in the biochemical clock. Ryanodine (up to 60 microM) did not affect the pacemaker frequency, which indicates that a ryanodine-sensitive store, if it exists, is not coupled to the biochemical clock. Electron microscopy showed that the smooth ER forms an extensive network of subsurface cisternae that is closely associated with large areas of the cytoplasmic face of the plasma membrane. These structures were the most extensive in interstitial cells of Cajal, slightly less in branching smooth muscle cells and far less in circular muscle cells. In summary, on the basis of these electrophysiological and morphological observations, we hypothesize that the Ca++ refilling cycle of the IP3-sensitive calcium stores associated with the plasma membrane determines the frequency of the pacemaker activity generated by the submuscular interstitial cells of Cajal-smooth-muscle network of the canine colon. PMID- 7473134 TI - Reye syndrome model in rats: protection against liver abnormalities by L carnitine and acetyl-L-carnitine. AB - The effects of L-carnitine (LCn) and acetyl-L-carnitine (AcLCn) were assessed on the liver alterations observed in Kilpatrick's model of Reye syndrome in rats; fasted rats were given lipopolysaccharide (LPS), 0.2 mg/kg i.p., 12 hr before they were sacrificed, plus acetylsalicylic acid (ASA), 50 mg/kg i.p., 11 hr before sacrifice; LCn or AcLCn were given twice, 500 mg/kg orally, 12 and 2 hr before sacrifice. LPS+ASA-treated rats showed a dramatic decrease of hepatic ketone bodies and acetyl-CoA and an increase of isobutyryl-CoA, isovaleryl-CoA and succinyl-CoA. Electron microscopy of LPS+ASA-treated rat liver showed a slight but significant alteration in mitochondrial inner structure. Because impairment of mitochondrial function in RS is associated with swelling, we investigated whether the microviscosity of mitochondrial lipids and the cholesterol-phospholipid ratio (CHOL/PL), were involved in the RS model used. Mitochondria from LPS+ASA-treated rats showed a decrease in lipid microviscosity, in CHOL/PL ratio and in CHOL/PL ratio of both inner and outer membrane fractions; these alterations suggested a general increase in membrane fluidity. LCn and AcLCn reversed the morphological alterations in mitochondria after LPS+ASA, observed by electron microscopy, the decrease in KB and the toxic increase in short-chain acyl-CoAs; AcLCn only reversed the decrease in acetyl-CoA. LCn and AcLCn prevented mitochondrial lipid alterations mainly in the inner membrane fraction.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7473135 TI - Effects of putative cannabinoid receptor ligands, anandamide and 2-arachidonyl glycerol, on immune function in B6C3F1 mouse splenocytes. AB - Anandamide (arachidonylethanolamide), isolated from the porcine brain, and 2 arachidonyl-glycerol (2-Ara-Gl), derived from the canine gut, are two recently identified putative endogenous cannabinoid receptor ligands. Both ligands have been reported to possess binding affinity for cannabinoid receptor subtypes, CB1 and CB2. The objective of the present studies was to investigate the immunomodulatory effects of both of these ligands in B6C3F1 mouse splenocytes. 2 Ara-Gl produced a marked and dose-related inhibition of the mixed lymphocyte response, anti-CD3 mAb-induced T-cell proliferation and LPS-induced B-cell proliferation, whereas having no inhibitory effect on phorbol-12-myristate-13 acetate/ionomycin-induced cell proliferation. Interestingly, the inhibitory effects by 2-Ara-Gl on proliferation were at least dependent in part on cell density. At high cell density, 2-Ara-Gl enhanced lymphoproliferation whereas exhibiting marked inhibitory activity at low cell density. Similarly, in vitro primary immunoglobulin M antibody-forming cell responses which are dependent on high cell density also were found to be enhanced by 2-Ara-Gl. Conversely, anandamide exhibited no inhibitory effects on cell proliferative responses to stimulation by anti-CD3 mAb, lipopolysaccharide or phorbol-12-myristate-13 acetate/ionomycin treatment. Anandamide also showed no effect on the in vitro sheep erythrocyte antibody-forming cell response. Although shown previously to markedly inhibit forskolin-stimulated cyclic AMP accumulation, 2-Ara-Gl exhibited no effect on basal adenylate cyclase activity in splenocytes. Additionally, anandamide showed negligible inhibitory effects at extremely high concentrations on forskolin-stimulated adenylate cyclase activity and no effect on basal adenylate cyclase activity in splenocytes.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7473136 TI - Disposition characteristics of recombinant human interleukin-11 after a bolus intravenous administration in mice. AB - The pharmacokinetics and disposition characteristics of recombinant human interleukin-11 (rhIL-11) after systemic administration of 10 to 1000 micrograms/kg was investigated in mice. After a bolus i.v. injection of 100 micrograms/kg of 111In-labeled rhIL-11, radioactivity disappeared rapidly from the circulation after a biexponential function. Plasma clearance profiles based on immunoreactivity and biological activity were identical to the disappearance pattern of radioactivity in the early phase after injection. In the range of 10 to 100 micrograms/kg, pharmacokinetic parameter estimates such as the T1/2 alpha and total body clearance were almost constant, suggesting that pharmacokinetics of rhIL-11 were linear within this dose range. 111In-labeled rhIL-11 distributed mainly to the kidney and liver; within 10 min, 40 and 20% of the dose, respectively, accumulated. On the other hand, the urinary excretion ratio was very small (ca., 1% of the dose), suggesting that rhIL-11 was cleared rapidly by glomeruler filtration followed by efficient tubular reabsorption. Pharmacokinetic analysis of the tissue distribution data demonstrated large kidney and hepatic uptake clearances. At the highest dose (1000 micrograms/kg), total body clearance significantly decreased, due primarily to saturation of hepatic uptake. These findings will provide useful information for the development of rhIL-11. PMID- 7473137 TI - Sensitization to the behavioral effect of kainic acid in the mouse is mediated by nitric oxide. AB - Kainic acid (KA)-sensitive receptors are located on primary afferent C-fibers. Behavioral sensitization to each of four repeated injections of KA appears to involve activation of primary afferent C-fibers based on its susceptibility to capsaicin pretreatment. Hyperalgesia, thought to involve transmission along C fibers, is sensitive to pharmacologic manipulation of nitric oxide (NO). We tested the hypothesis that KA activates C-fibers, either directly or indirectly, by a mechanism that involves NO. Pretreatment with N omega-nitro-L-arginine methyl ester (L-NAME), an inhibitor of NO synthesis, inhibited KA sensitization whereas D-NAME, the inactive isomer, failed to mimic this action. D-Arginine also inhibited sensitization to KA, whereas L-arginine, a NO precursor, was inactive when administered alone but reversed the inhibitory effect of L-NAME. Methylene blue, which inhibits guanylyl cyclase and NO synthase, attenuated KA sensitization, suggesting that cyclic GMP synthesis may also be involved in this phenomenon. Reduced hemoglobin, which sequesters NO in the extracellular space, attenuated KA sensitization, indicating that the effect of NO is brought about in structures adjacent to cells in which it is synthesized. This convergence of data is consistent with the mediation of behavioral sensitization to KA by NO. KA sensitization has been shown to involve an action of the NH2 terminus of substance P (SP) and NO may thus mobilize SP. Consistent with this, in the presence of SP(1-7), methylene blue was no longer able to inhibit sensitization to KA, suggesting that NO evokes, rather than results from, mobilization of SP.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7473138 TI - Nitric oxide mediation of cocaine-induced dopaminergic behaviors: ambulation accelerating activity, reverse tolerance and conditioned place preference in mice. AB - Intraperitoneal administration of N omega-nitro-L-arginine (L-NNA, 7.5 and 15 mg/kg), a nitric oxide synthase inhibitor, inhibited the cocaine-induced ambulation-accelerating activity and the L-NNA administered before and during the chronic injection of cocaine in mice blocked the development of reverse tolerance to the ambulation-accelerating effect of cocaine (15 mg/kg). Mice were rendered reverse tolerant to cocaine by a s.c. injection of cocaine (15 mg/kg) once daily for 6 days. The development of reverse tolerance was evidenced by an increased response to cocaine, and the inhibition of reverse tolerance was evidenced by a lesser ambulatory response. L-NNA (5 and 10 mg/kg) administered before and during cocaine conditioning inhibited the development of cocaine-induced conditioned place preference (CPP). Cocaine-induced CPP was developed by an injection of cocaine (15 mg/kg) once every other day for a 6-day period (3 times). The increase in postsynaptic dopamine receptor sensitivity was also blocked by L-NNA in both cocaine-induced reverse tolerant and CPP mice. The dopamine receptor sensitivity was increased in cocaine-induced reverse tolerant and CPP mice as evidenced by an enhanced ambulatory activity to apomorphine (2 mg/kg). Therefore, the present results suggest that these dopaminergic behaviors of cocaine may be mediated partially via the activation of the nitric oxide system and that the enhancement of postsynaptic dopamine receptor sensitivity may be an underlying common mechanism that mediates the cocaine-induced dopaminergic behaviors such as reverse tolerance and CPP. PMID- 7473139 TI - Supra-additive natriuretic synergism between bendroflumethiazide and furosemide in rats. AB - To examine the diuretic and natriuretic synergism between bendroflumethiazide (BFTZ) and furosemide (FUR), the acute diuretic and natriuretic response to BFTZ was compared when given alone, during acute and during chronic FUR infusion. Responses to diuretics were assessed in conscious, chronically instrumented rats and, to avoid confounding influences of diuretic-induced volume contraction, extracellular fluid volume was kept constant by a computer-driven servo-control technique. Two groups were pretreated with osmotic minipumps chronically infusing either FUR (0.25 mg/hr; group CF; n = 9) or vehicle (8% ethanolamine; group CV; n = 8) i.p. for 7 days before the experiment. During the experiment, two other groups received either acute infusion with FUR (0.25 mg/hr; group AF; n = 16) or acute infusion with vehicle (150 mM glucose; group AV; n = 11). After a 60-min control period, BFTZ (0.083 mg; 0.25 mg/hr) was administered for 90 min to all four groups. Whereas the diuretic and natriuretic response to BFTZ was similar in groups AF, AV and CV, a supra-additive response was observed in the CF group (vs. group CV: delta urine flow rate: +78%; delta urinary Na excretion: +69%). BFTZ had no effect on the fractional excretion of Li, which suggests that the supra additive effect was due to inhibition of an enhanced distal tubular Na reabsorption induced by chronic FUR administration. Because before BFTZ administration urinary Na excretion was 3-fold higher during acute than during chronic FUR infusion, an increased delivery of NaCl to the thiazide segment cannot by itself explain the exaggerated BFTZ response during chronic FUR treatment.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7473140 TI - Variability in human cytochrome P450 paclitaxel metabolism. AB - Formation of 6 alpha-hydroxypaclitaxel has been described as the primary biotransformation pathway for paclitaxel in vitro and in vivo, with additional formation of two other "minor" metabolites. Using a large group (n = 49) of human liver microsomes, and P450s heterologously expressed in cell lines, our aims were to elucidate the P450s responsible for and investigate variability in paclitaxel metabolite formation. Four metabolites of paclitaxel (6 alpha-hydroxypaclitaxel, metabolites B, C and A) were formed in vitro, via CYP2C8, 3A4, 3A4 and both 2C8 and 3A4, respectively. Although 6 alpha-hydroxypaclitaxel was predominant in the majority of livers, metabolites B and C (formed by CYP3A4) were predominant in 11/49 and 2/49 livers, respectively. Predominance of metabolite B over 6 alpha hydroxypaclitaxel was more likely in liver microsomes from donors known to be exposed to phenobarbital (P = .009), and tended to be more likely in diseased vs. normal livers (P = .047). Formation rates for 6 alpha-hydroxypaclitaxel, A, B, and C were lower in diseased liver vs. normal liver (P < .001). Rates of formation of metabolites B and C were highly correlated with each other (r2 = .91; P < .001) and with midazolam 4-hydroxylation (r2 = .87 & 0.86, respectively; P < .001). Inhibitor experiments suggest that typical CYP3A substrates/inhibitors (e.g., cyclosporin, epipodophyllotoxins) may significantly interact with paclitaxel in vivo. In a single patient in whom plasma samples were measured on two occasions, metabolite A (the dihydroxylate) was predominant, and systemic clearance of paclitaxel was lower in a course administered 1 day vs. 6 wk after a course of fluconazole therapy. We report that 6 alpha-hydroxypaclitaxel, formed via CYP2C8, is not the predominant paclitaxel metabolite in all individuals, and that CYP3A4 catalytic activity is important to overall paclitaxel metabolism in humans. PMID- 7473141 TI - Serotonin stimulates rapid increase of inositol 1,4,5-trisphosphate in ovine uterine artery: correlation with contractile state. AB - Serotonin (5-HT)-mediated inositol 1,4,5-trisphosphate (Ins(1,4,5)P3) synthesis and contractions were examined in isolated sheep uterine arteries. 5-HT stimulated a rapid increase of Ins(1,4,5)P3 production with the peak at 30 sec. The accumulation of Ins(1,4,5)P3 was transient and declined to a steady state slightly above the basal level at 4 min. The increase of inositol 1,3,4,5 tetrakisphosphate (Ins(1,3,4,5)P4) was also rapid, reaching the peak at 60 sec, and subsequently declining to the steady state at 4 min. Comparison of the time courses of 5-HT-induced Ins(1,4,5)P3 production with the force development indicated that increase of Ins(1,4,5)P3 content preceded the force development in the initial phasic component, but subsequently decreased, whereas the maximal tension was maintained. Consistent with the time courses, there was a nonlinear temporal relationship between Ins(1,4,5)P3 production and the force development measured simultaneously in the same tissue stimulated by 10 microM 5-HT. 5-HT stimulated Ins(1,4,5)P3 was concentration-dependent with EC50 of 0.48 microM. In accordance, 5-HT produced concentration-dependent contractions. The dissociation constant (KA) of 5-HT in the uterine artery was 0.52 +/- 0.08 microM. Plotting the relative responses as a function of the fractional receptor occupancy indicated a hyperbolic relationship for contractions, but a linear relationship for Ins(1,4,5)P3 production. Simultaneous measurement of contractions and Ins(1,4,5)P3 productions elicited by 5-HT (0.1-3 microM) revealed a significant linear correlation between these two events. The 5-HT-mediated Ins(1,4,5)P3 response was blocked by ketanserin (0.1 microM), but not by prazosin (0.1 microM). Pretreatment of tissues with pertussis toxin (200 ng/ml, 3 hr) failed to block 5-HT-induced inositol phosphates accumulation.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7473142 TI - HP-228, a novel synthetic peptide, inhibits the induction of nitric oxide synthase in vivo but not in vitro. AB - alpha-Melanocyte stimulating hormone has been shown to prevent endotoxin shock. A heptapeptide analog (HP-228) has recently been synthesized and shown to be an even more potent protective agent. Because the hypotensive and toxic actions of lipopolysaccharide (LPS) appear to involve the induction of type II nitric oxide synthase (iNOS), we have examined the actions of HP-228 on nitric oxide production using an endotoxemia model in conscious rats given E. coli LPS (5 mg/kg i.v.) and monitored for 6 h. A group of rats received HP-228 (30 micrograms/kg) 30 min before LPS. Using nitro L-arginine methyl ester-sensitive cGMP production as an estimate of nitric oxide synthase activity in aortic segments, ex vivo, we determined that LPS increases iNOS activity and that HP-228 pretreatment markedly reduces this response. Additionally, the rate of conversion of 3[H]-arginine to 3[H]-citrulline was significantly reduced in lung homogenates from HP-228-treated rats. HP-228 did not alter the activity of the constitutive nitric oxide synthase in aortic rings or in cerebella. In isolated rat aortic smooth muscle cells, LPS or interleukin-1 beta caused prominent rises in nitric oxide generated by iNOS. HP-228 did not antagonize the effect of these inducing agents. However, in these cells, plasma obtained from rats 1 h after administration of HP-228 prevented the induction of iNOS by both LPS and interleukin-1 beta. In conclusion, HP-228 prevents the in vivo induction of nitric oxide synthase by LPS.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7473143 TI - N-demethylation of amitriptyline in vitro: role of cytochrome P-450 3A (CYP3A) isoforms and effect of metabolic inhibitors. AB - Biotransformation of amitriptyline (AMI) to its demethylated product nortriptyline (NT) was studied in vitro with human liver microsomes from four different donors, preselected to reflect a range of metabolic rates. Reaction velocity versus substrate concentration was consistent with a sigmoid Vmax model. Vmax varied from 0.42 to 3.42 nmol/mg/min, Km from 33 to 89 microM AMI. Ketoconazole was a highly potent inhibitor of N-demethylation, with a mean Ki value of 0.11 +/- 0.013 microM (+/- S.D.), whereas quinidine (up to 50 microM), a CYP2D6 inhibitor, and alpha-naphthoflavone (up to 5 microM), a CYP1A2 inhibitor only at low concentrations, showed no effect. All selective serotonin reuptake inhibitors (SSRIs) tested had an inhibitory effect on the formation of NT, with mean Ki values of 4.37 (+/- 3.38) microM for sertraline, 5.46 (+/- 1.95) microM for desmethylsertraline, 9.22 (+/- 3.69) microM for fluvoxamine, 12.26 (+/- 5.67) microM for norfluoxetine, 15.76 (+/- 5.05) microM for paroxetine, and 43.55 (+/- 18.28) microM for fluoxetine. A polyclonal rabbit antibody against rat liver CYP3A1, in antibody/microsomal protein ratios varying from 1:1 to 10:1, inhibited N-demethylation of AMI to an asymptotic maximum of 60%. These results are consistent with several case reports describing impairment of AMI metabolism by SSRIs. Inhibition of AMI demethylation by low concentrations of ketoconazole and by anti-3A antibody supports an important role for CYP3A isoforms in mediating this reaction. PMID- 7473144 TI - Pharmacological characterization of GT-2016, a non-thiourea-containing histamine H3 receptor antagonist: in vitro and in vivo studies. AB - GT-2016, a non-thiourea-containing imidazole, has been developed as a histamine H3 antagonist. In vitro and in vivo studies in rats were conducted to characterize receptor selectivity and autoreceptor functionality for GT-2016. GT 2016 demonstrated high affinity (43.8 +/- 3.0 nM) and selectivity for the histamine H3 receptor in vitro. In vivo, GT-2016 (3, 10 and 30 mg/kg i.p. and p.o.) was shown to cross the blood-brain barrier and dose-dependently bind to cortical histamine H3 receptors. GT-2016 induced dose-dependent increases in histamine turnover at concentrations that exhibited significant histamine H3 receptor occupancy. Also, in vivo microdialysis experiments were conducted in awake, freely moving rats treated with GT-2016. GT-2016 (10 and 30 mg/kg i.p.) increased histamine release by approximately 75% above baseline within 1 hr, and elevated histamine release was observed for up to 2.5 hr after the higher dose. In contrast, GT-2016 was devoid of activity on histamine methyltransferase in vitro at concentrations up to 3 microM. Taken together, the results show that GT 2016 crosses the blood-brain barrier, binds to H3 receptors and increases the release of histamine in the cerebral cortex, consistent with blockade of presynaptic H3 autoreceptors. In summary, these findings allowed us to identify and characterize the in vitro and in vivo biochemical properties of a novel H3 receptor antagonist, GT-2016. PMID- 7473145 TI - Factors affecting serum protein binding of cocaine in humans. AB - The free (unbound) drug in serum is an important determinant of pharmacologic response. The present study was performed to more completely identify and evaluate factors affecting cocaine binding to human serum proteins. Protein binding was determined by ultrafiltration with [3H] cocaine. Cocaine binding parameters in serum from eight healthy volunteers were determined over a concentration range of 0.003 to 300 microM (0.001-100 micrograms/ml) and indicated cocaine binds to two classes of independent binding sites; one with high affinity [association constant (Ka) = 0.42 +/- 0.09 microM-1] and low capacity (N1 = 12.3 +/- 2.9 microM) and one with low affinity and high capacity (gamma = 0.41 +/- 0.05). Binding was concentration dependent with free fraction increasing from 0.16 +/- 0.05 to 0.68 +/- 0.02 over this concentration range. The binding capacity was significantly correlated with alpha-1-acid glycoprotein (AAG) concentration (r2 = 0.71, P = .0009). Binding studies were performed using AAG and human serum albumin (HSA) alone and together in phosphate buffer to determine the specific proteins responsible for cocaine binding. These studies revealed the binding of cocaine to AAG is potentiated by the presence of HSA as Ka for the first binding site increased from 0.08 microM-1 with AAG alone to 0.46 microM-1 with AAG combined with HSA 4 g/dl. Binding parameter estimates and cocaine free fraction in human serum and AAG 75 mg/dl plus HSA 4 g/dl in phosphate buffer were similar indicating that AAG and HSA are the principal binding proteins in serum.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7473146 TI - Effects of a selective DP receptor agonist (BW 245C) and antagonist (BW A868C) on the canine colonic epithelium: an argument for a different DP receptor? AB - Earlier studies showed that PGD2 produced both increases and decreases in short circuit current across the canine proximal colon. PGD2 metabolites had opposing effects: 11 beta-PGF2 alpha elicited only increases in Isc, and 13,14-dihydro-15 keto-PGD2 elicited only decreases. The stimulant effects involved FP receptors, but the receptors involved in mediating the inhibitory effects remained undefined. Here we show that the tissue, although it is capable of producing both PGD2 and PGE2, did not produce 11 beta-PGF2 alpha in measurable amounts. The selective DP receptor agonist BW 245C did not mimic the inhibitory effects of PGD2, producing only dose-dependent increases in short-circuit current. Further, these responses were not significantly inhibited by BW A868C. Cross desensitization experiments suggested that the stimulant effects of BW 245C involved the EP receptor. However, on a standard preparation (rabbit platelets), both PGD2 and BW 245C inhibited ADP-induced aggregation and were antagonized by BW A868C. 11 beta-PGF2 alpha had no effects. The decreases in short-circuit current across the canine colonic epithelium elicited by PGD2 and 13,14-dihydro 15-keto PGD2 are not mimicked by other prostanoids, nor by the selective agonist BW 245C, and thus appear to involve receptors other than the classical DP receptor. PMID- 7473147 TI - Antinociceptive effects of R(-)- and S(+)-flurbiprofen on rat spinal dorsal horn neurons rendered hyperexcitable by an acute knee joint inflammation. AB - The antinociceptive effects of the S(+)-enantiomer of flurbiprofen (potent inhibitor of cyclooxygenase) and the R(-)-enantiomer (500 times less potent) were investigated in the spinal cord of 20 anesthetized rats. In lumbar segments, 20 wide-dynamic-range dorsal horn neurons with knee joint input were recorded extracellularly. After induction of an acute inflammation in the knee joint by kaolin and carrageenan, the neurons developed hyperexcitability consisting of enhanced responses to stimuli applied to the inflamed knee and the noninflamed ankle and an expansion of receptive fields. Intravenous administration of R(-) flurbiprofen (1-9 mg/kg) and S(+)-flurbiprofen (0.3-9 mg/kg) at 5.5 to 8.5 h after kaolin, dose dependently reduced the neurons' responses to pressure applied to the inflamed knee (18 of 18 neurons) and the noninflamed ankle (17 of 17 neurons) and paw (8 of 8 neurons). R(-)-flurbiprofen decreased the receptive field size in 8 of 16 neurons, S(+)-flurbiprofen in 10 of 16 neurons. The suppressive effects started 3 to 6 min and reached a maximum 9 to 15 min after i.v. administration. S(+)-flurbiprofen was more potent than the R(-)-enantiomer. When injected directly into the knee joint, S(+)-flurbiprofen (50 and 80 micrograms), but not the R(-)-enantiomer (100 and 180 micrograms) reduced the hyperexcitability in 12 of 12 neurons. These results suggest a central site of antinociceptive action for R(-)-flurbiprofen and S(+)-flurbiprofen and an additional peripheral site for S(+)-flurbiprofen. The doses used in these experiments did not produce any sedative effects in rats subjected to behavioral testing. PMID- 7473148 TI - Effects of intrathecal and intracerebroventricular buprenorphine on a C-fiber reflex in the rat. AB - A C-fiber reflex elicited by electrical stimulation within the territory of the ipsilateral sural nerve, was recorded from the biceps femoris muscle in anesthetized rats. The temporal evolution of the reflex was studied using a constant level of stimulus intensity (3 x threshold) and recruitment curves were built by varying stimulus intensity from 0 to 7 x threshold. Intrathecal doses of buprenorphine in the 0.2 to 10 micrograms range, elicited a facilitation of the C fiber reflex in a dose-dependent manner. A large dose (100 micrograms) depressed but did not block the reflex. Intracerebroventricular doses of buprenorphine in the 0.1 to 10 micrograms range, facilitated the C-fiber reflex. A higher dose (100 micrograms) elicited a biphasic effect: depressive when the stimulus intensity was weak and facilitatory when the stimulus intensity was strong. It is concluded that the antinociceptive properties of buprenorphine cannot be related to a direct or indirect depressive spinal effect. In terms of spinal and supraspinal effects of buprenorphine, it is likely that buprenorphine facilitates the C-fiber reflex via a supraspinal mechanism that acts on sensory and/or motor components of the reflex arc although the depression of the reflex involves a spinal mechanism. The lipophilic properties of buprenorphine could explain a substantial diffusion from its spinal injection site to the brain. From a clinical standpoint, this study confirms that intrathecal administration of buprenorphine is an inadequate way of accessing spinal opioid receptors. PMID- 7473149 TI - Allosteric interactions at L-type calcium channels between FPL 64176 and the enantiomers of the dihydropyridine Bay K 8644. AB - Functional interactions between the enantiomers of the dihydropyridine 1,4 dihydro-2,6-dimethyl-5-nitro-4-[2-(trifluoromethyl)-phenyl]-3-pyridi ne carboxylic acid methyl ester (Bay K 8644) and the benzoylpyrrole methyl 2,5 dimethyl-4-[2(phenylmethyl)benzoyl]-H-pyrrole-3-carboxylate (FPL 64176) were investigated on L-type Ca++ channels in guinea pig ileal longitudinal smooth muscle. The effects of these drugs, when applied individually, were as described in earlier studies. For instance, both (-)-(S)-Bay K 8644 and FPL 64176 caused concentration-dependent contraction, which is consistent with Ca++ channel activation, whereas (+)-(R)-Bay K 8644 gave concentration-dependent relaxation, which is consistent with Ca++ channel inhibition. The activities of the different drugs were dependent on the extracellular levels of KCI. When applied in combination, however, the responses evoked were not those predicted from the effects of the drugs applied individually. Contractions produced by FPL 64176 (25 nM to 1 microM) were abolished in the presence of 100 nM (-)-(S)-Bay K 8644 but were potentiated by 10 to 150 nM (+)-(R)-Bay K 8644 and inhibited by 1 microM (+) (R)-Bay K 8644. Conversely, contractile responses to (-)-(S)-Bay K 8644 were abolished by 100 nM FPL 64176. In the presence of 1 microM FPL 64176, however, ( )-(S)-Bay K 8644 gave concentration-dependent relaxation of the muscle, which is consistent with Ca++ channel inhibition.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7473150 TI - Cardiovascular responses to cigarette smoke exposure in restrained conscious rats. AB - The effect of exposure to cigarette smoke on cardiovascular function was examined in conscious, restrained Sprague-Dawley rats. Rats were exposed to 3, 6 and 9 puffs of either air or cigarette smoke during the "break in" period and to 10 puffs on the day of the experiment (day 4). HR, cardiac output and mean arterial pressure were recorded continuously throughout the experimental period. Rats exposed previously to cigarette smoke generated from either low-nicotine (0.16 mg/cig.) or high-nicotine (2.45 mg/cig.) cigarettes showed a dose-related decrease in HR in response to restraint stress. In addition, exposure to cigarette smoke produced a further decrease in HR and cardiac output and an increase in mean arterial pressure. This effect by cigarette smoke was dose dependent (dependent on the cigarette nicotine content) and was antagonized by intra-arterial pretreatment with the nicotinic antagonists mecamylamine and hexamethonium and also with the ganglionic blocker chlorisondamine. Intra arterial pretreatment with atropine methyl bromide blocked the bradycardia in response to both restraint stress and cigarette smoke. Furthermore, pretreatment with an arginine vasopressin antagonist, d(CH2)5Tyr(Me)arginine vasopressin, significantly attenuated the increase in mean arterial pressure and total peripheral resistance and the decrease in HR and cardiac output due to cigarette smoke. On the other hand, pretreatment with the opioid receptor antagonist naloxone had no effect on cardiovascular parameters in response parameters in response to cigarette smoke. These results implicate arginine vasopressin, in addition to the activation of both sympathetic and parasympathetic systems, in mediating cardiovascular responses to cigarette smoke. PMID- 7473151 TI - Depletion of hepatic 3'-phosphoadenosine 5'-phosphosulfate (PAPS) and sulfate in rats by xenobiotics that are sulfated. AB - Sulfation is considered a high-affinity but low-capacity conjugation mechanism that is limited by the availability of 3'-phosphoadenosine 5'-phosphosulfate (PAPS), the cosubstrate for sulfation. Salicylamide, phenol and 1-naphthol are all known substrates for the sulfation reaction. This study was conducted to determine whether the xenobiotics that are sulfated when administered to rats will lower hepatic PAPS and its precursor, sulfate. Urinary sulfate excretion was reduced 85% to 95% by these compounds. Hepatic PAPS was reduced 73%, 39%, and 87% by salicylamide, phenol and naphthol, respectively, 2 hr after administration of 2 mmol/kg. These compounds also decreased serum sulfate concentrations by 45% to 86% and lowered hepatic sulfate concentrations. In summary, these studies demonstrate that salicylamide, phenol and 1-naphthol lower hepatic PAPS and sulfate concentrations, as well as serum sulfate concentrations. These findings imply that increased sulfation, as a result of the sulfation of xenobiotics, results in depletion of hepatic PAPS concentrations, possibly because the utilization of PAPS by the sulfotransferases exceeds its generation via sulfate activation. Thus the capacity-limited sulfation of high dosages of xenobiotics appears to be due to the reduced availability of hepatic PAPS, which in turn is limited by the availability of sulfate. PMID- 7473152 TI - Coronary and hemodynamic effects of S 16257, a new bradycardic agent, in resting and exercising conscious dogs. AB - The effects of (7,8-dimethoxy 3-[3-([(1S)-(4,5-dimethoxybenzocyclobutan-1 yl)methyl]methylamino) - propyl]1,3,4,5-tetrahydro-2H-benzapin 2-one hydrochloride) (S 16257), a new bradycardic agent acting through inhibition of the hyperpolarization-activated I(f) current in the sino-atrial node, on arterial blood pressure, cardiac output, left ventricular pressure and dP/dt, epicardial coronary artery diameter and coronary blood flow velocity were investigated at graded doses (0.1 to 1 mg/kg) and compared to those of propranolol (1 mg/kg) and saline in chronically instrumented conscious dogs. At rest, S 16257 induced a dose-dependent bradycardia that was significant at 0.5 (-16% +/- 3%) and 1 mg/kg (-23% +/- 3%). At 0.5 mg/kg, S 16257 decreased both resting HR and treadmill exercise-induced tachycardia to a similar extent as propranolol. However, in contrast to propranolol, S 16257 (0.5 mg/kg) did not affect the increase in mean coronary blood flow velocity and the decrease in coronary vascular resistance observed during a control exercise performed under saline. In addition, S 16257 did not affect resting epicardial coronary artery diameter and only attenuated its increase during exercise, whereas propranolol reduced it at rest and maintained a significant constriction of these large vessels throughout the exercise period. Propranolol, but not S 16257, decreased left ventricular dP/dt at each stage of exercise.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7473153 TI - Different effects of indomethacin and nabumetone on prostaglandin-mediated gastric responses to central vagal activation in rats. AB - Intracisternal injection of a stable thyrotropin-releasing hormone (TRH) analog increases gastric prostaglandins release and mucosal resistance to injury through central vagal pathways. The effects of two nonsteroidal anti-inflammatory drugs, indomethacin (INDO) and nabumetone on intracisternal injection of various doses of TRH-induced gastric acid secretion and changes in mucosal resistance were investigated in urethane-anesthetized rats. Doses of INDO (5 mg/kg) and nabumetone (13.75 mg/kg) producing similar acute anti-inflammatory response in the carrageenin-induced paw edema were injected i.p. in all studies. INDO potentiated the acid secretion induced by intracisternal injection of TRH at 25, 50 and 200 ng by 5.1-, 1.9- and 1.4-fold, respectively, whereas nabumetone did not modify the secretory response to TRH. Moderate erosions were observed in 100% of rats treated with the combination of INDO and TRH (200 ng) whereas no erosions were observed when TRH or INDO were given alone or TRH in combination with nabumetone. TRH at 7 ng reduced mucosal damage induced by intragastric administration of ethanol (60%, 1 ml/kg) by 63%. The mucosal protective action of TRH was abolished by INDO but not altered by nabumetone pretreatment. These data indicate that at comparable anti-inflammatory doses, nabumetone, unlike INDO, neither blocks the protection against ethanol injury induced by low doses of TRH injected intracisternally nor potentiates the gastric acid secretion or lesions induced by higher dose of TRH. We speculate that these differences reflect reduced inhibition of gastric prostaglandins by nabumetone. PMID- 7473154 TI - Role of second messengers in agonist up-regulation of 5-HT2A (5-HT2) receptor binding sites in cerebellar granule neurons: involvement of calcium influx and a calmodulin-dependent pathway. AB - The present study was undertaken to examine the involvement of 5-HT2A receptor mediated second messengers in the agonist-induced up-regulation of 5-HT2A receptors in cerebellar granule cells. Stimulation of these cells with a 5-HT2A/5 HT2C agonist, (+/-)-(2, 5-dimethoxy-4-iodophenyl)-2-aminopropane for 16 hr resulted in a marked increase in [3H]ketanserin binding to 5-HT2A receptors in intact cells. (+/-)-(2, 5-dimethoxy-4-iodophenyl)-2- aminopropane up-regulated, but not basal levels of 5-HT2A binding sites, were largely attenuated by actinomycin D and cycloheximide, suggesting an essential role for de novo RNA and protein synthesis in this up-regulation process. This receptor up-regulation was 5-HT2A receptor-mediated but was unaffected by short-term pretreatment with phorbol dibutyrate to attenuate (+/-)-(2, 5-dimethoxy-4-iodophenyl)-2 aminopropane-induced phosphoinositide hydrolysis or by treatment with staurosporine to inhibit protein kinase C. In contrast, blockade of Ca2+ channels by LaCl3 or SK&F 96365 and depletion of extracellular Ca2+ by EGTA preferentially decreased the up-regulated 5-HT2A receptor levels, suggesting a role of Ca2+ influx in the receptor up-regulation. The (+/-)-(2, 5-dimethoxy-4- iodophenyl)-2 aminopropane up-regulation was also prevented by inhibitors of calmodulin, calmidazolium and W-7. Moreover, the effect of (+/-)-(2, 5-dimethoxy-4 iodophenyl)-2-aminopropane was blocked by KN-62, a selective Ca2+/calmodulin kinase inhibitor and by H-7 and staurosporine at concentrations high enough to be nonselective for a specific type of protein kinase.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7473155 TI - Pinacidil relaxes porcine and human coronary arteries by activating ATP-dependent potassium channels in smooth muscle cells. AB - We investigated the effect of the potassium channel opener pinacidil on ATP dependent K+ channels (KATP) in the relaxation of porcine and human coronary arteries by means of isometric contraction experiments in arterial rings. We also measured whole cell currents in freshly isolated porcine and human coronary artery vascular smooth muscle cells with patch clamp. We first characterized serotonin-induced precontractions in our vessels and proved that the contractions were mediated by Ca2+ influx through voltage-dependent Ca2+ channels. Similarly, we observed that serotonin-induced contractions were strongly enhanced by small K(+)-induced depolarizations. Pinacidil completely relaxed rings preconstricted with 5 microM serotonin and produced dose-dependent relaxations of 5 microM serotonin-preconstricted rings, with an IC50 of 1.26 microM. Similar results were observed (IC50 = 1.15 microM) when the endothelium was removed. The KATP blocker glibenclamide (3 microM), inhibited pinacidil-induced relaxations (5-10 microM) by approximately 25% although the KATP blocker tetrapentylammonium (10 microM), inhibited pinacidil-induced (5-10 microM) relaxations completely. Pinacidil 10 microM had only a minimal effect on rings precontracted with a 50 mM external K+ concentration (IC50 = 60 microM). Porcine and human arterial rings did not differ qualitatively in their responses. Moreover, in the patch clamp experiments pinacidil (1 microM and 20 microM) induced a large, nonrectifying, outward current in both human and porcine cells. The reversal potential was close to the K+ equilibrium potential, suggesting an induction of pinacidil-activated K+ current. The pinacidil-induced (1 microM) current was strongly inhibited by glibenclamide (3 microM). These data show that the relaxation of porcine and human coronary arteries by pinacidil is primarily induced by an opening of KATP in smooth muscle cells. Furthermore, the vasorelaxant effect of pinacidil is not endothelium dependent. PMID- 7473156 TI - Anticonvulsant tolerance and withdrawal characteristics of benzodiazepine receptor ligands in different seizure models in mice. Comparison of diazepam, bretazenil and abecarnil. AB - The use of benzodiazepines (BDZs) in the long-term treatment of epilepsy is greatly restricted by their capacity to induce tolerance and dependence. Thus, the development of new BDZ-related therapeutic agents should be directed by strategies that minimize tolerance- and dependence-inducing properties. Experimental procedures used to determine the success of such strategies often rely on a single assay procedure (e.g., one seizure model), which might lead to false predictions. Furthermore, the different types of tolerance, i.e., "pharmacological" (metabolic or functional) and "behavioral" ("learned" or "contingent"), are often not dealt with in such studies. This prompted us to compare the chronic anticonvulsant efficacy and withdrawal characteristics of diazepam and two novel BDZ receptor ligands, i.e., the partial agonist bretazenil and the subtype-selective agonist abecarnil, in different seizure models in mice. Myoclonic, clonic and tonic seizures were induced by i.v. infusion of pentylenetetrazol and by transcorneal or transauricular application of electrical stimuli. Prolonged administration of diazepam (5 mg/kg twice daily for 6 days) resulted in marked anticonvulsant effects on myoclonic, clonic and tonic seizure thresholds at the onset of treatment, but pronounced tolerance developed rapidly during subsequent treatment. The time course and extent of tolerance was similar with most seizure models. Tolerance characteristics were not affected by study design, i.e., use of separate or the same animals for each seizure induction, indicating that learned or contingent tolerance was not significantly involved under these experimental conditions. After termination of treatment with diazepam, significant seizure threshold decreases were determined, indicating withdrawal hyperexcitability in response to physical dependence. During prolonged administration of abecarnil (10 mg/kg twice daily for 6 days), some anticonvulsant tolerance was seen with electroshock seizures, but not with pentylenetetrazol seizures; no withdrawal hyperexcitability was determined upon termination of treatment. Bretazenil (10 mg/kg twice daily for 6 days) produced no tolerance in any of the seizure models, but a significant decrease in electroshock seizure threshold was seen in the withdrawal period. The data indicate that tolerance and withdrawal characteristics of BDZ receptor partial and subtype-selective agonists in mice depend on the experimental model used, whereas the influence of the experimental protocol is less critical in the case of a full BDZ receptor agonist such as diazepam. PMID- 7473157 TI - Inhibition of lipopolysaccharide-induced pulmonary edema by isozyme-selective phosphodiesterase inhibitors in guinea pigs. AB - There is a need for pharmacological agents for the treatment of pulmonary edema associated with the adult respiratory distress syndrome. Therefore, we examined the effects of isozyme-selective cyclic AMP phosphodiesterase (cAMP PDE) inhibitors, as well as aminophylline and dexamethasone, on the pulmonary edema, protein leakage into the airways and airway neutrophilia induced by aerosolized lipopolysaccharide (LPS) in intact guinea pigs. Twenty-four hours after LPS exposure lung wet/dry weight ratios increased from 4.9 +/- 0.004 to 5.8 +/- 0.02. Rolipram (PDE4 selective), CI-930 (PDE3 selective), aminophylline and dexamethasone (given p.o. 1 hr before and 4 hr after LPS exposure) inhibited pulmonary edema formation with ED50 values of 1.7, 0.5, 31 and 2.8 mg/kg, respectively. Maximum inhibition occurred with rolipram at 10 mg/kg (70 +/- 17%), CI-930 at 10 mg/kg (101 +/- 4%), aminophylline at 50 mg/kg (88 +/- 14%) and dexamethasone at 3 mg/kg (64 +/- 6%). Denbufylline and milrinone also inhibited pulmonary edema formation at 10 mg/kg i.p., supporting the inhibition of PDE4 and PDE3 as the mechanisms of action of rolipram and CI-930, respectively. Rolipram, CI-930, aminophylline and dexamethasone (at maximum doses for inhibiting pulmonary edema) inhibited the 3-fold increase in bronchoalveolar lavage albumin concentration 24 hr after LPS exposure (42 +/- 14%, 98 +/- 2%, 70 +/- 9% and 53 +/- 13%, respectively). However, none of these compounds (at maximum doses for inhibiting pulmonary edema) inhibited the corresponding 400-fold increase in lavage neutrophil counts.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7473158 TI - Prostaglandin E2 contracts vascular smooth muscle and inhibits potassium currents in vascular smooth muscle cells of rat tail artery. AB - There is evidence to suggest that PGE2 plays an important role in the regulation of vascular smooth muscle tone. To determine the cellular basis of this action, we studied the effect of PGE2 on force in helical muscle strips from rat tail artery. PGE2 evoked a sustained contractile response. The contractile response was concentration-dependent, with an EC50 value of 9.6 microM. Patch-clamp studies were conducted to investigate the effects of PGE2 on K channels in isolated vascular smooth muscle cells from rat tail artery. Current-clamp studies showed that PGE2 (1 microM) depolarized the membrane by 15.9 +/- 1.3 mV. Under voltage-clamp conditions, a voltage-dependent, delayed outward rectifier K current was generated by stepwise depolarization from a holding potential of -80 mV. The current, which was activated at -45 to -40 mV and showed almost no inactivation, was inhibited by 45% using 10 mM TEA. PGE2 inhibited the outward K current in a concentration-dependent manner, with EC50 values of 3.5 microM and 4.9 microM in primary and subcultured cells, respectively. The PGE2 receptor antagonist sodium meclofenamate abolished the PGE2-induced K current inhibition. Furthermore, the intracellular application of guanosine 5'-O(-)[2 thiodiphosphate] (GDP beta S), a G protein inhibitor, and pretreatment of the cells with cholera toxin prevented the PGE2-induced inhibition, whereas application of pertussis toxin did not.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7473159 TI - Galanin receptors in human basal forebrain differ from receptors in the hypothalamus: characterization using [125I]galanin (porcine) and [125I]galantide. AB - Galanin, a 29-amino acid peptide, is uniquely distributed in human basal forebrain and may play a role in cholinergic cell dysfunction in Alzheimer's disease. We report a detailed evaluation of galanin receptors in human basal forebrain (67 +/- 12 years) and hypothalamus (67 +/- 15 years) with radioligand binding techniques. The binding of [125I]galanin (porcine) (agonist) or [125I]galantide [GAL (1-3)-substance P (5-11)-NH2] (putative antagonist) saturated in 2 hr, and only 15% to 30% of either radioligand was removed in the presence of unlabeled peptide. [125I]Galanin or [125I]galantide binding in basal forebrain revealed similar Bmax values, with [125I]galanin having a higher affinity for the galanin receptor. In contrast, [125I]galanin showed a lower affinity and labeled 42% more receptors than [125I]galantide in the hypothalamus. Differences were noted in competition studies of galanin and galanin chimeric peptides (M15, M35, M40 and C7) between [125I]galanin and [125I]galantide binding and in both regions. M35, M40 and C7 showed high affinity for galanin receptors in the hypothalamus with Hill coefficients close to unity, whereas in the basal forebrain these peptides competed differently. 5'-Guanylylimidodiphosphate reduced the specific binding of either radioligand in both regions. Based on the derived data, both radioligands irreversibly bind with high affinity and act as agonists at galanin receptors in human basal forebrain and hypothalamus. Galanin and galanin chimeric peptides compete differently for galanin receptors depending on the radioligand and region tested, suggesting subtype differences. PMID- 7473160 TI - Effects of (-)-S-2,8-dimethyl-3-methylene-1-oxa-8-azaspiro[4,5]decane L-tartrate monohydrate (YM796), a novel muscarinic agonist, on disturbance of passive avoidance learning behavior in drug-treated and senescence-accelerated mice. AB - Effects of YM796 (-)-S-2,8-dimethyl-3-methylene-1-oxa-8-azaspiro[4,5]decane L tartrate monohydrate; a novel muscarinic agonist, were observed on disturbance of passive avoidance learning behavior in drug- (protein synthesis inhibitor and anticholinergic drugs) treated and senescence-accelerated mice in comparison with those of a muscarinic agonist (AF102B) and acetylcholinesterase inhibitors (E2020 (1-benzyl-4-[(5,6-dimethoxy-1-indanone-2-yl) methyl] piperidene hydrochloride), NIK247 [9-amino-2,3,5,6,7,8-hexahydro-1H-cyclopenta(b)-quinoline monohydrate hydrochloride], THA (9-amino-1,2,3,4-tetrahydroacridine) and physostigmine). All tested drugs administered before training significantly prolonged the shortened latency of step-through induced by the protein synthesis inhibitor cycloheximide (150 mg/kg s.c.). This shortened latency was also significantly prolonged when YM796 was administered immediately after training, but not when administered before the test trial. The ameliorating effect of YM796 on the impairment in learning behavior by cycloheximide was significantly suppressed by pirenzepine (0.1 micrograms/mouse i.c.v.). When administered before training, all test drugs prolonged the shortened latency of step-through induced by treatment with the anticholinergic drugs [scopolamine (1 mg/kg s.c.) and hemicholinium-3 (0.3 microgram/mouse i.c.v.)], suggesting that they ameliorated the impairment of learning behavior. This shortened latency in scopolamine-treated mice was also significantly prolonged by YM796, AF102B, E2020, NIK247 and physostigmine when administered immediately after training, but not when administered before the test trial. The pharmacological actions of YM796 administered immediately after training and before the test trial in hemicholinium-3-treated mice were similar to those in scopolamine-treated mice.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7473161 TI - Pharmacological characterization of LY303870: a novel, potent and selective nonpeptide substance P (neurokinin-1) receptor antagonist. AB - LY303870 [(R)-1-[N-(2-methoxybenzyl)acetylamino]-3-(1H-indol-3-yl)-2-[N-(2-(4- (piperidin-1-yl)piperidin-1-yl)acetyl)amino]propane] is a new, potent and selective nonpeptide neurokinin-1 (NK-1) receptor antagonist. LY303870 bound selectively and with high affinity to human peripheral (Ki = 0.15 nM) and central (Ki = 0.10 nM) NK-1 receptors. LY303870 inhibited [125I]substance P (SP) binding to guinea pig brain homogenates with similar affinity; however, it had approximately 50-fold lesser affinity for rat NK-1 sites. The less active enantiomer, LY306155 [(S)-1-[N-(2-methoxybenzyl)acetylamino]-3-(1H-indol-3-yl)-2 [N-(2-(4- (piperidin-1-yl)piperidin-1-yl)acetyl)amino]-propane], was 1,000- to 15,000-fold less potent in all the species examined. LY303870 antagonized in vitro NK-1 receptor effects as demonstrated by blockade of SP-stimulated phosphoinositide turnover in UC-11 MG human astrocytoma cells (Ki = 1.5 nM) and interleukin-6 secretion from U-373 MG human astrocytoma cells (Ki = 5 nM). In addition, LY303870 inhibited SP-induced rabbit vena cava contractions (pA2 = 9.4) with high (50,000-fold) selectivity vs. NK-2 or NK-3 receptor-mediated responses. In vivo, LY303870 inhibited [Sar9,Met(O2)11]-SP induced guinea pig bronchoconstriction (ED50 = 75 micrograms/kg i.v.) and pulmonary microvascular leakage in the bronchi (ED50 = 12.8 micrograms/kg i.v.) and trachea (ED50 = 18.5 micrograms/kg i.v.). Therefore, LY303870 is a potent and selective NK-1 receptor antagonist in vitro and in vivo. The use of LY303870 will facilitate a better understanding of NK-1 receptors in physiological processes. PMID- 7473162 TI - Cardiovascular effects of the novel dual inhibitor of neutral endopeptidase and angiotensin-converting enzyme BMS-182657 in experimental hypertension and heart failure. AB - Combined neutral endopeptidase (NEP) and angiotensin-converting enzyme (ACE) inhibition produces greater acute hemodynamic effects than either treatment alone. We investigated whether BMS-182657 (BMS), which bears inhibitory activities against both NEP and ACE, elicited similar enhanced effects. BMS inhibited NEP and ACE, in vitro (IC50 = 6 and 12 nM, respectively) and the pressor response to Ang I in rats. In deoxycorticosterone acetate (DOCA)-salt hypertensive rats sensitive to NEP inhibition but not to ACE inhibition, BMS at 100 mumol/kg i.v. lowered mean arterial pressure (MAP) from 180 +/- 6 to 151 +/- 5 mm Hg. In sodium-depleted, spontaneously hypertensive rats (SHR) sensitive to ACE inhibition but not to NEP inhibition, BMS at 100 mumol/kg p.o. lowered MAP from 151 +/- 4 to 123 +/- 5 mm Hg. Cardiomyopathic hamsters with heart failure were administered vehicle or one of the following (30 mumol/kg i.v.): the ACE inhibitor enalaprilat; the NEP inhibitor SQ-28603; or BMS. Enalaprilat and SQ 28603 had minimal hemodynamic effects. BMS decreased left ventricular end diastolic pressure by 12 +/- 2 and 10 +/- 1 mm Hg and left ventricular systolic pressure by 27 +/- 2 and 23 +/- 3 mm Hg at 30 and 60 min, respectively (P < .05 vs. each other group). These changes were associated with a 40% increase in cardiac output, a 47% decrease in peripheral vascular resistance and a lowering of MAP by 21 +/- 3 mm Hg at 60 min (P < .05 vs. each other group). There were no significant differences in the changes in heart rate or left ventricular stroke work index among the four groups. Hence, BMS-182657 is a dual inhibitor of NEP and ACE, is antihypertensive irrespective of the activity of the renin angiotensin system and has acute hemodynamic effects in hamsters with heart failure greater than those produced by selective inhibition of NEP or ACE. The NEP and ACE inhibitory activities of BMS-182657 act synergistically and mimic the interaction resulting from combining selective inhibitors of these enzymes. PMID- 7473163 TI - NMDA antagonist properties of the putative antiaddictive drug, ibogaine. AB - Both anecdotal reports in humans and preclinical studies indicate that ibogaine interrupts addiction to a variety of abused substances including alcohol, opiates, nicotine and stimulants. Based on the similarity of these therapeutic claims to recent preclinical studies demonstrating that N-methyl-D-aspartate (NMDA) antagonists attenuate addiction-related phenomena, we examined the NMDA antagonist properties of ibogaine. Pharmacologically relevant concentrations of ibogaine produce a voltage-dependent block of NMDA receptors in hippocampal cultures (Ki, 2.3 microM at -60 mV). Consistent with this observation, ibogaine competitively inhibits [3H]1-[1-(2-thienyl)-cyclohexyl]piperidine binding to rat forebrain homogenates (Ki, 1.5 microM) and blocks glutamate-induced cell death in neuronal cultures (IC50, 4.5 microM). Moreover, at doses previously reported to interfere with drug-seeking behaviors, ibogaine substitutes as a discriminative stimulus (ED50, 64.9 mg/kg) in mice trained to discriminate the prototypic voltage-dependent NMDA antagonist, dizocilpine (0.17 mg/kg), from saline. Consistent with previous reports, ibogaine reduced naloxone-precipitated jumping in morphine-dependent mice (ED50, 72 mg/kg). Although pretreatment with glycine did not affect naloxone-precipitated jumping in morphine-dependent mice, it abolished the ability of ibogaine to block naloxone-precipitated jumping. Taken together, these findings link the NMDA antagonist actions of ibogaine to a putative "antiaddictive" property of this alkaloid, its ability to reduce the expression of morphine dependence. PMID- 7473164 TI - U-90042, a sedative/hypnotic compound that interacts differentially with the GABAA receptor subtypes. AB - U-90042 is a structurally novel compound that has comparable affinities for binding to three recombinant subtypes of the gamma-aminobutyric acidA receptor: alpha 1 beta 2 gamma 2, alpha 3 beta 2 gamma 2 and alpha 6 beta 2 gamma 2. The relatively high affinity for the alpha 6 beta 2 gamma 2 subtype is similar to the benzodiazepine (BZ) partial inverse agonist Ro 15-4513 and different from BZ sedative/hypnotics such as diazepam and zolpidem. In the present study, U-90042 (3 mg/kg i.p.) suppressed locomotor activity and impaired rotarod performance in mice. These effects were not antagonized by flumazenil. The sedative effect was further confirmed in rats (10 mg/kg i.p.) and monkeys (1 mg/kg p.o.) by an increase of behavioral sleep and a corresponding electroencephalographic frequency spectral shift. Unlike the BZ hypnotics, U-90042 (10 mg/kg i.p.) produced no amnesia in the one-trial passive avoidance response in mice but antagonized diazepam-induced amnesia. In rats trained to discriminate an injection of diazepam from saline, U-90042 produced predominantly vehicle appropriate responses, even at depressant doses. The in vivo diazepam-antagonist effect of U-90042 is consistent with its low intrinsic activity and diazepam antagonism at the gamma-aminobutyric acidA alpha 1 beta 2 gamma 2 and alpha 3 beta 2 gamma 2 receptor subtypes. The receptor mechanism for the sedative/hypnotic effect is not clear at this time. PMID- 7473165 TI - Effect of oleanolic acid on hepatic toxicant-activating and detoxifying systems in mice. AB - We have previously shown that oleanolic acid (OA) protects mice against the hepatotoxicity of carbon tetrachloride, acetaminophen, bromobenzene, thioacetamide, furosemide, phalloidin, colchicine, cadmium, D-galactosamine and endotoxin. This study was designed to examine whether OA modulates hepatic toxicant-activating and detoxifying systems as a means of protection. Mice were treated with OA (100 and 200 mumol/kg s.c.) for 3 days, and liver microsomes and cytosols were prepared 24 hr after the last dose. OA produced a dose-dependent reduction in liver microsomal cytochrome P450 (P450) levels (25-37%) and cytochrome b5 (15-21%) content, but had no effect on NADPH-cytochrome c reductase activity. OA treatment also decreased several P450 enzyme activities, such as coumarin 7-hydroxylation (45%), 7-pentoxyresorufin O-dealkylation (35%), 7 ethoxyresorufin O-dealkylation (25%) and chlorzoxazone 6-hydroxylation (20%). Treatment of mice with OA decreased caffeine N3-demethylation (40%), but had no effect on caffeine 8-hydroxylation. OA treatment decreased testosterone 6 alpha- and 15 alpha-hydroxylation (40-50%) and androstenedione formation (35%), but slightly increased testosterone 1 alpha/beta-, 2 beta- and 6 beta-hydroxylation. Consistent with enzyme activities, OA decreased the amounts of mouse liver CYP1A and CYP2A enzymes, but had no appreciable effect on CYP3A enzymes, as determined by immunoblotting with antibodies against rat P450 enzymes. OA treatment slightly increased liver glutathione (GSH) content and the activity of GSH S-transferases toward 1-chloro-2,4-dinitrobenzene, but had no effect on GSH peroxidase and GSH reductase. The activities of superoxide dismutase and DT-diaphorase were unaffected by OA treatment. At the high dose of OA, catalase activity was decreased by 20%.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7473166 TI - Increased function of voltage-dependent Ca++ channels and Ca(++)-activated K+ channels in resting state of femoral arteries from spontaneously hypertensive rats at prehypertensive stage. AB - The present study examined the possible role of voltage-dependent Ca++ channels (VDCs) and Ca(++)-activated K+ (KCa) channels in the regulation of resting tone of arteries from spontaneously hypertensive rats (SHR) at a prehypertensive stage. Differences in the effects of agents that interact with these channels were assessed in endothelium-denuded strips of femoral arteries isolated from 4 week-old SHR and age-matched normotensive Wistar-Kyoto rats (WKY). Systolic blood pressures at this age were not significantly different between SHR and WKY. The arterial strips from SHR maintained a myogenic tone in the resting state; that is the resting tone in the SHR artery was abolished when either the bathing solution was replaced with a Ca(++)-free solution or 10(-7) M nifedipine was added. Studies using 1- or 5-min pulse labeling of the arteries with 45Ca showed that the resting Ca++ influx was significantly increased in SHR when compared with WKY, and this increase in SHR was abolished by 10(-7) M nifedipine. In strips preloaded with fura-PE3, the addition of 3 x 10(-6) M verapamil to resting muscles decreased the resting cytosolic Ca++ level and caused a relaxation. These effects of verapamil were more evident in SHR than in WKY. The addition to the strips of charybdotoxin and iberiotoxin, blockers of large conductance KCa channels, caused a concentration-dependent contraction, which was significantly greater in SHR than in WKY.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7473167 TI - Chronic neurosteroid treatment decreases the efficacy of benzodiazepine ligands and neurosteroids at the gamma-aminobutyric acidA receptor complex in mammalian cortical neurons. AB - The effect of chronic 5 alpha-pregnane-3 alpha-ol-20-one (5 alpha 3 alpha; neurosteroid) treatment was investigated on the gamma-aminobutyric acid (GABA), 5 alpha 3 alpha, and ligands that bind to the benzodiazepine (BZ) site on GABA induced [36Cl-]influx in intact cultured mammalian cortical neurons. Chronic 5 alpha 3 alpha treatment (1 microM; 5 days) decreased the efficacy of GABA, because its Emax (maximal response) value was decreased, whereas the EC50 (potency) value was not altered. Chronic 5 alpha 3 alpha treatment also decreased the Emax value of BZ agonists like diazepam to potentiate GABA-induced [36Cl-] influx, and decreased the -Emax (maximal inhibitory response) value of inverse agonists like methyl-6,7-dimethoxy-4-ethyl-beta-carboline-3'-carboxylate to inhibit GABA-induced [36Cl-] influx, whereas not altering their EC50/IC50 values. Furthermore, chronic 5 alpha 3 alpha treatment decreased the Emax value of 5 alpha 3 alpha to potentiate GABA-induced [36Cl-] influx, without altering its EC50 value. The decreased efficacy of GABA and 5 alpha 3 alpha were reversed by concomitant exposure of the neurons to R 5135 (3 alpha-hydroxy-16-imino-5 beta-17 androstan-11-one; a competitive GABA antagonist). Taken together, these findings suggest that chronic 5 alpha 3 alpha treatment produces decreased efficacy of GABA, ligands that bind to the BZ site, and neurosteroids at the GABAA-BZ receptor complex. The decreased efficacy is heterologous in nature and involves mediation via the GABAA receptor site. PMID- 7473168 TI - Critical role for glucocorticoid receptors in stress- and ethanol-induced locomotor sensitization. AB - Locomotor sensitization, the augmentation of the locomotor-activating effects of stimuli with repeated exposure, is being evaluated as a partial model for several phenomena including drug addiction. Alteration of dopaminergic systems has been found in sensitized animals and dopamine neurotransmission appears to be crucial for the expression of sensitized behaviors. However, stress hormones, which are released after exposure to many of the stimuli that produce sensitization, may also be involved in the development of this phenomenon. Corticosterone appears to be important in the development of amphetamine sensitization and glucocorticoid receptors (GR) have been hypothesized to mediate this effect. The purpose of these experiments was first, to determine whether repeated restraint stress sensitizes DBA/2J mice to the activating effect of ethanol (EtOH), and second, to explore the role of GR in stress- and EtOH-induced sensitization with the GR antagonist, RU 38486. This antagonist was administered before restraint or i.p. EtOH (1.5 g/kg) on each of 10 consecutive days of pretreatment. In addition, plasma corticosterone levels were determined at various points throughout the pretreatment period and on test days. The results demonstrated that 10 consecutive days of 2-hr restraint sensitized mice to EtOH's locomotor stimulating effect. Both stress- and EtOH-induced sensitization were attenuated by administration of RU 38486 during the pretreatment phase.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7473170 TI - KR-30450, a newly synthesized benzopyran derivative, activates the cardiac ATP sensitive K+ channel. AB - KR-30450 (2-(2"(1",3"-dioxolone)-2-methyl-4-(2'-oxo-1'-pyrrollidinyl) -6- nitro 2H-1-benzopyren) is a newly synthesized benzopyran derivative. We examined the effect of KR-30450 on the action potential duration of isolated rat papillary muscle and on the ATP-sensitive K+ channel (KATP) activity in single rat ventricular myocytes with 3 M KCl-filled conventional microelectrode and patch clamp techniques. KR-30450 (10(-7) approximately 10(-5) M) reduced the action potential duration in a concentration-dependent manner and this was inhibited by 3 microM glibenclamide, suggesting that KATP was involved. In cell-attached patches, KR-30450 (10(-5) M) in the pipette activated the KATP which was closed by 3 microM glibenclamide. In inside-out patches, the effects of KR-30450 on KATP activity were examined before and after run-down of the channel. Before run-down, KR-30450 increased the KATP activity only in the presence of ATP and shifted the [ATP]i-KATP activity relationship to the right. After run-down, KR-30450 did not affect the KATP activity either in the presence or absence of 3 mM UDP, but increased the UDP-induced KATP activity in the presence of 1 mM ATP-gamma-S. From these results, we conclude that KR-30450 antagonizes the inhibitory effect of ATP on the KATP in a competitive manner. These effects of KR-30450 are similar to those of ER-001533 and HOE-234, but different from those of pinacidil and lemakalim. PMID- 7473169 TI - EUK-8, a synthetic superoxide dismutase and catalase mimetic, ameliorates acute lung injury in endotoxemic swine. AB - Reactive oxygen metabolites are believed to be important mediators of sepsis- or lipopolysaccharide (LPS)-induced adult respiratory distress syndrome. EUK-8 is a novel, synthetic, low-molecular-weight salen-manganese complex that exhibits both superoxide dismutase and catalase activities in vitro. We hypothesized that treatment with EUK-8 would ameliorate pulmonary dysfunction in a porcine model of LPS-induced adult respiratory distress syndrome. At T = -18 h, pigs received an intravenous priming dose of LPS (20 micrograms/kg). Anesthetized ventilated swine were randomized to receive 1) no further treatment (n = 5); 2) LPS (250 micrograms/kg from T = 0 to 60 min, n = 6); 3) LPS and a low dose of EUK-8 (10 mg/kg bolus at T = -15 min and 1 mg/kg.h from T = 0 to 240 min, n = 6) or 4) LPS and a higher dose of EUK-8 (10-mg/kg bolus and 3 mg/kg.h, n = 6). Treatment with EUK-8, particularly at the higher dose, significantly attenuated many of the features of LPS-induced acute lung injury, including arterial hypoxemia, pulmonary hypertension, decreased dynamic pulmonary compliance and pulmonary edema. LPS caused an increase in lung tissue malondialdehyde content that was abrogated in both EUK-8-treated groups. EUK-8 treatment had no effect on circulating plasma levels of tumor necrosis factor-alpha, thromboxane B2 or 6 keto-prostaglandin F1 alpha. We conclude that EUK-8 prevents many of the manifestations of LPS-induced adult respiratory distress syndrome in pigs by detoxifying reactive oxygen metabolites without affecting the release of other important proinflammatory mediators. PMID- 7473171 TI - Effects of photodegradation products of nifedipine: the nitroso-derivative relaxes contractions of the rat aortic strip induced by norepinephrine and other agonists. AB - Effects of four photodegradation products of nifedipine (nitroso-, nitro-, azoxy- and N,N'-dioxide-derivatives) were studied in the rat aortic strip suspended in the bath medium. In the CaCl2-free bath medium containing 40 mM KCl, CaCl2 (2.5 mM) induced contraction, which was relaxed by nifedipine (-log [EC50] = 8.66 M) and all the four photodegradation products (-log[EC50] = 4.15-4.82 M). The EC50 values for all these substances were not affected by denuding the vessel of endothelium. In the bath medium containing KCl (15 mM), Bay K-8644 (2 x 10(-6) M) induced contraction, which was relaxed by nifedipine (10(-9)-10(-6) M) and its four photodegradation products (3 x 10(-6)-10(-4) M). Norepinephrine (10(-7) M) induced contraction, which was relaxed by the nitroso-derivative (10(-4) M) noncompetitively (pIC50 = 4.37) but not by three other photodegradation products. Both methylene blue and hemoglobin had no effect on the relaxation produced by the nitroso-derivative. Phorbol 12-myristate 13-acetate induced contraction, which was inhibited completely by the nitroso-derivative (10(-4) M) but only by 42% by nifedipine (10(-4) M). Prostaglandin F2 alpha (3 x 10(-6) M), endothelin-1 (10(-9) M) and 5-hydroxytryptamine (10(-6) M) also induced contraction, which was relaxed effectively by the nitroso-derivative (10(-4) M) but not by any of the three other photodegradation products.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7473173 TI - Plasma digoxin immunoreactivity and arterial potassium relaxation after quinapril therapy in hypertensive rats. AB - Plasma digitalis-like substance and altered function of arterial Na+,K(+)-ATPase have both been linked with elevated blood pressure, but the influence of antihypertensive therapy on these factors remains unknown. Therefore, we treated spontaneously hypertensive rats and normotensive Wistar-Kyoto rats with the angiotensin-converting enzyme inhibitor quinapril for 10 weeks. The therapy markedly reduced blood pressure and plasma digoxin immunoreactivity, and it normalized the elevated plasma Na+:K+ ratio in the hypertensive animals. Relaxations of endothelium-denuded denervated arterial rings induced by return of potassium to the organ bath upon precontractions elicited by potassium-free solution were used to evaluate the function of vascular Na+,K(+)-ATPase. The rate of potassium relaxation was faster in quinapril-treated hypertensive rats and in both Wistar-Kyoto groups than in the hypertensive controls. Potassium relaxation was also effectively inhibited by the Na+,K(+)-ATPase inhibitor ouabain in all groups. In addition, arterial contractions to potassium chloride and relaxations to nitroprusside were examined. The contractions to lower concentrations of potassium chloride (20 mM) were enhanced in untreated hypertensive rats when compared with the other groups, although the maximal responses were corresponding in all groups. The time to reach base-line tension after washout of potassium chloride (125 mM) and the relaxations to nitroprusside did not differ in the study groups. In conclusion, the present results showed that long-term angiotensin-converting enzyme inhibition in parallel reduced plasma digoxin-like factor, enhanced arterial potassium relaxation (probably reflecting enhanced function of Na+,K(+)-ATPase) and normalized plasma Na+:K+ ratio in this type of genetic hypertension. PMID- 7473172 TI - A drug discrimination analysis of the actions of novel serotonin1A receptor ligands in the rat using the 5-HT1A receptor agonist, 8-hydroxy-2-(di-n propylamino)tetralin. AB - In the present study, using a two-lever drug discrimination procedure, we characterized the effects of a series of chemically-diverse, novel 5-HT1A receptor agonists and antagonists in rats trained to discriminate the serotonin (5-HT)1A receptor agonist, 8-hydroxy-2-(di-N-propylamino) tetralin (8-OH-DPAT) (0.31 mg/kg, i.p.) from saline. In analogy to the 5-HT1A receptor agonists, ipsapirone (a pyrimidinylpiperazine) and flesinoxan (a benzodioxane), the arylpiperazine derivatives, WY-48,723 and WY-50,324, as well as the methoxynaphtylpiperazines, S 14506 and S 14671, substituted 100% for the discriminative stimulus (DS) effects of 8-OH-DPAT, with the latter two displaying remarkable potency [50 effective doses (ED50s): 1.25, 0.34, 0.05, 0.69, 0.009 and 0.006 mg/kg, s.c., respectively]. In contrast, an additional pyrimidinylpiperazine, zalospirone, failed to fully mimick the training drug (maximal effect: 40%) even at a dose markedly disrupting response rates (2.5 mg/kg, s.c.). The potency of agonists in generalizing to 8-OH-DPAT correlated significantly (P < .05) with their affinity at rat hippocampal 5-HT1A receptors in vitro (r = .78), and with their potency to induce hypothermia in the rat (r = .96). S 15535 and S 15931, novel benzodioxopiperazines possessing mixed 5-HT1A autoreceptor agonist/postsynaptic 5-HT1A receptor antagonist properties, antagonized (approximately 75%) the discriminative stimulus (DS) properties of 8 OH-DPAT (ED50s: 6.9 and 0.97 mg/kg, s.c., respectively), although their structural analogue, S 14489, reduced by only 50% the action of 8-OH-DPAT. The 5 HT1A receptor antagonists, (+/-)-pindolol (-70%; ED50: 0.65 mg/kg, s.c.), (-) alprenolol (-67%; ED50: 7.1) and NAN-190 (-80%; ED50: 1.5), all blocked the 8-OH DPAT DS. Likewise, several novel antagonists at 5-HT1A autoreceptors and postsynaptic 5-HT1A receptors; the benzoisothiazolpiperazine, SDZ 216-525 (-83%; ED50: 0.64), the aryloxyalkylamine, (-)-tertatolol (-83%; ED50: 7.7) and the methoxyphenylpiperazine, (+)-WAY 100,135 (-80%; ED50: 17.0), antagonized the 8-OH DPAT cue. Antagonist potency correlated significantly with affinity at 5-HT1A receptors (r = .83) and potency for antagonism of 8-OH-DPAT-induced hypothermia (r = .83). Antagonists showed only variable and not significant (P > .05) generalization rates (13-50%) when tested alone.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 7473174 TI - Muscarine induces an anomalous inhibition of synaptic transmission in rat auditory thalamic neurons in vitro. AB - Muscarinic receptor-mediated inhibition of central synaptic transmission was studied in a monosynaptic pathway connecting the inferior colliculus and the auditory thalamus in in vitro rat brain explants. Extra- and intracellular synaptic responses were recorded by sharp electrode and whole-cell patch clamp techniques in the ventral nucleus of the medial geniculate body after electrical stimulation of the brachium of the inferior colliculus. Stimulation of tectal afferents evoked either a high-frequency burst or a single-spike synaptic response in ventral geniculate neurons. Bath application of muscarinic receptor agonists abolished responses consisting of a high-frequency burst, but not responses consisting of a single spike. In the majority of single-spike cells muscarinic agonists often induced a synaptic facilitation. The burst blocking effect was mimicked by a moderate elevation of extracellular potassium. Intracellular recordings showed that the burst synaptic responses similar to that recorded extracellularly were induced by an excitatory postsynaptic potential. This synaptic potential, by first activating a low-threshold spike, was able to evoke a burst of sodium spike discharges. Muscarinic agonists caused a slow membrane depolarization that inactivated the low-threshold spike, leading to a blockade of the burst response. This mechanism is tentatively termed here as EPSP LTS decoupling. Our results therefore support the hypothesis that part of the muscarinic receptor-mediated synaptic inhibition previously reported in anesthetized animal preparations in vivo represents a membrane depolarization rather than pre- or postsynaptic inhibition. PMID- 7473175 TI - (+)-N-allylnormetazocine enhances N-acetyltransferase activity and melatonin synthesis: preliminary evidence for a functional role of sigma receptors in the rat pineal gland. AB - In the present study, to evaluate the role that sigma receptors play in the physiology of the pineal gland, we assessed the effects of the sigma receptor ligand (+)-N-allylnormetazocine on the gland activity during either the day or the night. As compared to saline, (+)-N-allylnormetazocine enhanced the physiological increases in both pineal N-acetyltransferase (NAT) activity and melatonin content at night, but it did not affect the biosynthetic activity of the gland during the day. Moreover, (+)-N-allylnormetazocine potentiated the enhancement of NAT activity and pineal melatonin content induced by isoproterenol administration during the day. The nocturnal stimulation of pineal NAT activity and melatonin levels by (+)-N-allylnormetazocine was prevented by pretreatment with rimcazole, a specific sigma receptor antagonist. These results demonstrate that sigma receptor activation by (+)-N-allylnormetazocine is not able, by itself, to stimulate pineal melatonin production, whereas it potentiates the biosynthetic activity of the pineal gland when this is stimulated noradrenergically. PMID- 7473176 TI - Effects of methylprednisolone and 21-aminosteroids on mitogen-induced interleukin 6 and tumor necrosis factor-alpha production in human peripheral blood mononuclear cells. AB - The compounds U-74389G (16-desmethyl tirilazad) and U-74500A are two of the novel series of nonglucocorticoid 21-aminosteroids (or lazaroids) which mimic the high dose neuroprotective pharmacology of the glucocorticoid methylprednisolone (MP) in the injured CNS. Despite structural analogies to MP, it has been shown previously for a variety of endpoints that lazaroids are devoid of classical glucocorticoid effects. Our objective here was to measure the immunosuppressive effects of these lazaroids directly. Specifically, we have compared the in vitro effects of MP, U-74389G, and U-74500A on the mitogen-induced cytokine production in human peripheral blood mononuclear cells, which is known to be very sensitive and perhaps the most clinically relevant parameter reflecting immunomodulation. We show that, in contrast to the glucocorticoid MP, both lazaroids at therapeutically relevant concentrations have no significant inhibitory effects on stimulated interleukin-6 and tumor necrosis factor-alpha production, neither via residual glucocorticoid receptor-mediated activities nor via direct physicochemical effects on cellular membranes. These results strongly support the view that lazaroids lack glucocorticoid activities, but rather exert their tissue protective effects via mechanisms that are independent of glucocorticoid-receptor binding. PMID- 7473177 TI - Moexipril, a new angiotensin-converting enzyme (ACE) inhibitor: pharmacological characterization and comparison with enalapril. AB - The pharmacodynamic profile of the new angiotensin-converting enzyme (ACE) inhibitor moexipril and its active diacid, moexiprilat, was studied in vitro and in vivo. In vitro, moexiprilat exhibited a higher inhibitory potency than enalaprilat against both plasma ACE and purified ACE from rabbit lung. Upon oral administration of moexipril (10 mg/kg/day) to spontaneously hypertensive rats, plasma angiotensin II concentration decreased to undetectable levels, plasma ACE activity was inhibited by 98% and plasma angiotensin I concentration increased 8.6-fold 1 h after dosing. At 24 h, plasma angiotensin I and angiotensin II concentrations had returned to pretreatment levels, whereas plasma ACE activity was still inhibited by 56%. Four-week oral administration of moexipril (0.1-30 mg/kg/day) to spontaneously hypertensive rats lowered blood pressure and differentially inhibited ACE activity in plasma, lung, aorta, heart and kidney in a dose-dependent fashion. Equidose treatment (10 mg/kg/day) with moexipril and enalapril over 4 weeks led to comparable decreases in blood pressure, inhibition of plasma ACE and reduction of plasma angiotensinogen and to a similar attenuation of the pressor responses to angiotensin I and potentiation of the depressor responses to bradykinin. In contrast, ACE inhibition in aorta, heart and lung was significantly greater with moexipril than with enalapril, whereas in the kidney both drugs inhibited ACE activity to a similar extent. In summary, moexipril is an orally active ACE inhibitor that is comparable to enalapril in potency and duration of antihypertensive activity. The results of the present study demonstrate that 1) the antihypertensive potency of a given ACE inhibitor cannot be predicted from its in vitro characteristics and 2) the degree of blood pressure reduction does not correlate with tissue ACE inhibition. PMID- 7473179 TI - Regulation of striatal cyclic-3',5'-adenosine monophosphate accumulation and GABA release by glutamate metabotropic and dopamine D1 receptors. AB - In this study, the regulation of striatal cyclic-3',5'-adenosine monophosphate (cAMP) formation and GABA release by dopamine D1 and metabotropic glutamate receptors (mGluR) was studied in brain slices. In the absence of adenosine A2 receptor blockade, the mGluR agonist, 1-aminocyclopentane-1S,3R-dicarboxylic acid (1S,3R-ACPD) stimulated cAMP accumulation through a pertussis toxin-insensitive mechanism that could be blocked by L-serine-o-phosphate, but not by L(+)-2-amino 3-phosphonopropionic acid. However, in the presence of the adenosine antagonist, 3-isobutyl-1-methylxanthine, 1S,3R-ACPD had no significant effect on basal cAMP, but it inhibited cAMP formation stimulated by the D1 agonist, SKF 38393. This inhibitory response was prevented by pertussis toxin pretreatment and mimicked by L(+)-2-amino-3-phosphonopropionic acid, but it was unaffected by L-serine-o phosphate. Thus, 1S,3R-ACPD was determined to activate distinct mGluRs in the striatum that mediate either inhibition or activation of cAMP accumulation, with the latter effect being dependent on the activation of adenosine A2 receptors. A potential physiological role for the interaction between the D1 and adenosine dependent stimulatory metabotropic receptor was sought by examining this interaction on striatal GABA release. SKF 38393 and 1S,3R-ACPD together were found to potentiate striatal GABA release induced by 15 mM K+. The potentiation was blocked by the D1 antagonist, SCH 23390. However, this effect was only partially mimicked by a high concentration of forskolin (100 microM) and was not blocked by L-serine-o-phosphate, thereby suggesting that the stimulatory mGluR does not mediate this potentiation.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7473178 TI - Closed- and open-state binding of 4-aminopyridine to the cloned human potassium channel Kv1.5. AB - The effect of 4-aminopyridine (4-AP) on membrane and gating currents of Kv1.5 channels was studied in a human cell line. The rank order of block was cell attached > whole cell >> outside-out macropatches, which suggested that 4-AP blocked channels from the cytosolic face. Stimulation after exposure to 4-AP during rest resulted in a "supernormal" current immediately after the onset of depolarization, followed by open channel block during maintained depolarization. 4-AP remained trapped in closed channels, and unblock after drug washout required depolarization. At higher concentrations, 4-AP induced channel block by binding to closing or nonconducting channels. This effect could be reduced by hyperpolarization and higher pulsing rates. Block of Kv1.5 channels under steady state conditions was correlated with a dose-dependent reduction in gating charge movement. The normalized voltage dependence of gating was shifted to more negative potentials by 4-AP. A model incorporating both open and closed channel binding of 4-AP reproduced data, including the "supernormal" current, by shifting the Po - V curve slightly to the left. The results show that 4-AP can both enhance and block Kv1.5 current and suggest that 4-AP can bind with different affinities to more than one site on Kv1.5 channels. PMID- 7473180 TI - Functional correlates of dopamine D3 receptor activation in the rat in vivo and their modulation by the selective antagonist, (+)-S 14297: 1. Activation of postsynaptic D3 receptors mediates hypothermia, whereas blockade of D2 receptors elicits prolactin secretion and catalepsy. AB - Using [125I]-iodosulpride as a radioligand, the novel naphthofurane, (+/-)-S 11566 [(+/-)-[7-(N,N-dipropylamino)-5,6,7,8-tetra-hydro- naphtho(2,3b)dihydro,2,3 furane]) showed a marked preference for human, recombinant D3 as compared with D2 receptors stably transfected into Chinese hamster ovary cells (Kis = 24/529 nM). This activity resided in its (+)-eutomer, (+)-S 14297 (13/297 nM) as compared with its (-)-distomer, (-)-S 17777 (406/3544 nM). In contrast, (+)-AJ 76 manifested only a mild 2-fold preference for D3 sites (70/154 nM), whereas haloperidol and six additional antagonists showed a mild (2-7-fold) preference for D2 sites. As concerns agonists, (+)-7-OH-DPAT, (+/-)-CGS 15855A, quinelorane, (-)-quinpirole and N-0434 displayed a preference (6-40-fold) for D3 receptors, whereas piribedil showed a slight, 2-fold, preference for D2 sites (243/126 nM). (+)-S 14297 showed low (> 1.0 microM) affinity at rat D1 and D2 sites and at cloned, human D4 and D5 receptors and only low affinity (145 to > 10,000 nM) at all other sites examined. In vivo, administered s.c., (+)-7-OH-DPAT, CGS 15855A, quinelorane, (-)-quinpirole and N-0434 potently evoked hypothermia. Across all (8) agonists tested, potency correlated significantly with affinity at D3 sites (r = .84, P < .001) but not D2 sites (r = .50, P > .05). (+)-S 14297 (0.16-1.25 mg/kg, s.c.) blocked the induction of hypothermia by (+)-7-OH-DPAT, CGS 15855A and (-)-quinpirole, but not by the alpha 2-adrenergic agonist, clonidine, without influencing core temperature alone. In contrast, (-)-S 17777 (10.0 mg/kg, s.c.) was only partially active. Across all (9) antagonists, potency for inhibition of (+)-7-OH-DPAT-induced hypothermia correlated more strongly with affinity at D3 (r = .96, P < .001) than D2 (r = .75, P < .02) sites. Whereas haloperidol and the other antagonists provoked prolactin secretion and elicited catalepsy, (+)-S 14297 and (+/-)-S 11566 at doses of up to 10.0 and 40.0 mg/kg, s.c., respectively, were not significantly effective (P > .05). Across all antagonists, potency for eliciting prolactin secretion and catalepsy correlated better with affinity at D2 (r = .95 and .96) than D3 (r = .76 and .91) sites. In conclusion, these data demonstrate that the novel naphtofurane, (+)-S 14297, is a selective ligand (antagonist) at dopamine D3 receptors and suggest that their activation mediates hypothermia in the rat.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 7473181 TI - Functional correlates of dopamine D3 receptor activation in the rat in vivo and their modulation by the selective antagonist, (+)-S 14297: II. Both D2 and "silent" D3 autoreceptors control synthesis and release in mesolimbic, mesocortical and nigrostriatal pathways. AB - The preferential dopamine (DA) D3 versus D2 receptor agonist, (+)-7-OH-DPAT, dose dependently decreased DA synthesis in the nucleus accumbens, olfactory tubercles, striatum and frontal cortex. This action was potently mimicked by several other high-potency D3 agonists: CGS 15855A, (-)-quinpirole, quinelorane and N-0434. In contrast, piribedil, which displays a mild preference for D2 sites, was less active. Across eight agonists, potency for inhibition of DA synthesis correlated more potently to affinity at D3 (r = .82 +/- .04) than D2 receptors (r = .60 +/- .06, P < .05). Correlations were also marked to potency for induction of a further D3-mediated response, hypothermia (r = .93 +/- .02). The novel and selective D3 versus D2 antagonist, (+/-)-S 11556, attenuated the action of (+)-7 OH-DPAT in each structure. This action was shared by its active (+)-eutomer, (+) S 14297, whereas its inactive (-)-distomer, (-)-S 17777, was ineffective. (+)-S 14297 similarly attenuated the inhibitory action of CGS 15855A and (-)-quinpirole upon DA synthesis, whereas it failed to modify inhibition of striatal DA synthesis by the alpha 2-adrenergic receptor agonist, clonidine. As compared with the D2/D3 receptor antagonist, haloperidol, neither (+/-)-S 11566 nor (+)-S 14297 modified DA turnover upon administration alone. Furthermore, across (nine) antagonists, potency in facilitating DA synthesis more powerfully correlated to affinity at D2 (r = .94 +/- .01) than D3 (r = .73 +/- .01) sites (P < .01). Correlations were also marked to potency for induction of catalepsy (r = .91 +/- .01) and prolactin secretion (r = .89 +/- .01) but not for antagonism of (+)-7-OH DPAT-induced hypothermia (r = .60 +/- .01). In freely moving rats, (+)-7-OH-DPAT dose-dependently reduced dialysate concentrations of DA in the nucleus accumbens and contralateral striatum: this action was potently mimicked by CGS 15855A, but only weakly so by piribedil. (+)-S 14297 markedly attenuated the action of (+)-7 OH-DPAT, whereas (-)-S 17777 was inactive. In contrast, haloperidol completely blocked the action of (+)-7-OH-DPAT. Finally, in distinction to haloperidol, upon administration alone, (+)-S 14297 did not significantly enhance the release of DA. In conclusion, these data suggest that D3 (auto)receptors control synthesis and release of DA in dopaminergic pathways innervating the limbic system, cortex and striatum.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 7473182 TI - Telemetry monitoring of hemodynamic effects induced over time by adenosine agonists in spontaneously hypertensive rats. AB - Using the telemetry system in spontaneously hypertensive rats (SHR), we evaluated the hemodynamic effects of four adenosine analogs having different selectivity for A1 and A2a adenosine receptor subtypes. The selective A2a agonists, 2-hexynyl 5'-N-ethylcarboxamidoadenosine (2HE-NECA) and 2-[4-(2-carboxyethyl) phenethylamino]-5'-N-ethylcarboxamidoaden osi ne (CGS 21680), the selective A1 agonist, 2-chloro-N6-cyclopentyl-adenosine (CCPA) and the nonselective agonist, 5'-N-ethyl-carboxamidoadenosine (NECA), were administered i.p. to conscious spontaneously hypertensive rats. For comparison, the calcium channel blocker, felodipine, was included. CCPA and 2HE-NECA were tested also by the oral route. Systolic and diastolic blood pressure and heart rate were recorded every 5 min for 24 hr after drug administration. Data were analyzed using the curve fitting model recently elaborated. All compounds caused dose-dependent antihypertensive effects. The i.p. dose inducing a decrease of 50 mm Hg (ED50) was calculated for both systolic and diastolic blood pressure. As regards diastolic blood pressure, ED50 values were: CCPA, 0.019 (0.013-0.027) mg/kg, 2HE-NECA, 0.009 (0.002-0.03) mg/kg, CGS 21680, 0.155 (0.084-0.246) mg/kg, NECA, 0.008 (0.004-0.016) mg/kg and felodipine, 5.16 (4.18-7.18) mg/kg. At equiactive doses, the antihypertensive effects of adenosine agonists were shorter lasting [t1/2 for DBP were: CCPA, 54 (44-76) min, 2HE-NECA, 57 (46-71) min, CGS 21680, 45 (21-94) min, NECA, 61(38-97) min] than those of felodipine [t1/2 = 233 (182-274) min]. After oral administration (0.3, 1 and 3 mg/kg), hypotension induced by 2HE-NECA was longer lasting than that of CCPA. 2HE-NECA, CGS 21680 and felodipine caused tachycardia.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7473183 TI - Toxicodynamic analysis of cough and inflammatory reactions by angiotensin converting enzyme inhibitors in guinea pig. AB - Angiotensin-converting enzyme (ACE) inhibitors are one of the first drugs of choice for the treatment of hypertension. However, there have been many reports of persistent chronic dry cough and inflammatory skin reactions (rash and/or angioedema, etc.) induced by ACE inhibitors. In this study, in order to evaluate the cough and inflammatory reaction, we measured the number of citric acid induced coughs and the intradermal inflammation with ovalbumin in guinea pigs consecutively treated with ACE inhibitors (lisinopril, enalaprilat and imidapril) for 3 days. The number of citric acid-induced coughs and the inflammatory responses were significantly enhanced by treatment with lisinopril and enalaprilat, whereas imidapril produced no change in either response. These results correspond to the frequency of adverse effects in clinical practice, which suggests that imidapril has the least ability to induce the inflammatory skin response and cough. Furthermore, the enhancement produced by the ACE inhibitors in the number of coughs and the inflammatory responses were significantly reduced by pretreatment with indomethacin (prostaglandin synthesis inhibitor). This finding suggests that PGs at least participate in the mechanism for ACE inhibitor-induced cough and inflammatory skin response. PMID- 7473184 TI - Peritubular paraquat transport in isolated renal proximal tubules. AB - To better understand the characteristics of peritubular transport of organic cations (OCs), the uptake of the polyvalent OC dimethylbipyridinium (paraquat) and the structurally similar monovalent OC 1-methyl-4-phenylpyridinium (MPP+) was measured in suspensions of rabbit renal proximal tubules. Compared to the uptake of MPP+, the uptake of paraquat across the peritubular membrane was a low affinity, low capacity carrier-mediated process with a Jmax of 0.52 +/- 0.19 nmol.mg of protein.-1 min-1 and a Km of 162 +/- 25 microM. The uptake of MPP+ was a carrier-mediated process with a measured Jmax and Km of 1.8 +/- 0.09 nmol.mg of protein.-1min-1 and 28 +/- 8 microM, respectively. To determine whether paraquat is a substrate for the monovalent OC pathway, the effect of unlabeled MPP+ and tetraethylammonium (TEA) on paraquat uptake was examined. A 1 mM concentration of the monovalent OC MPP+ and TEA reduced the uptake of [14C]paraquat and [3H]MPP+ by approximately 30 and 90%, respectively, whereas 1 mM paraquat had no effect on [3H]MPP+ or [14C]TEA uptake. Thus, MPP+, but not paraquat, appears to interact with the monovalent OC transporter. On the other hand, the polyvalent OC substrates, including the polyamines putrescine and spermine, the herbicide diquat and the divalent hexamethonium bromidehydrate had no effect on either paraquat or MPP+ uptake. However, the synthetic polyamine methylglyoxal bis(guanyl-hydrazone)dihydrochloride (MGBG; 1 mM) reduced both paraquat and MPP+ uptake (by 60 and 90%, respectively). The ability of MGBG, unlike the other polyvalent substrates, to interact with paraquat transport may be related to structural similarities in the relative location of the two charged nitronium moieties in paraquat and MGBG.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7473185 TI - Membrane permeability and lipophilicity in the isolated perfused rat liver: 5 ethyl barbituric acid and other compounds. AB - The distribution kinetics of 5-ethyl barbituric acid (EBA) has been examined in the rat liver. The isolated in situ liver (n = 4) was perfused at a constant rate (15.1 +/- 0.2 ml/min, mean +/- S.D.) with protein-free Krebs bicarbonate medium in a single-pass mode. [14C]Sucrose (extracellular reference) and [14C]EBA were injected separately as bolus doses into the portal vein. The outflow data were analyzed using the axial dispersion model. The one-compartment dispersion model adequately described the data for sucrose, with a dispersion number (DN) of 0.25 +/- 0.04 and a volume of distribution (VH) of 0.14 +/- 0.01 ml/g liver. The two compartment dispersion model, which incorporates a cellular permeability barrier, provided a better description of the EBA outflow data. The estimated VH, influx and efflux rate constants and permeability-surface area product (PS) for EBA were 0.37 +/- 0.04 ml/g liver, 0.028 +/- 0.004 sec-1, 0.019 +/- 0.001 sec-1 and 3.4 +/ 0.5 ml/min, respectively. Despite low hepatocyte membrane permeability, the DN value for EBA (0.28 +/- 0.03) was not significantly different from that of sucrose, which supports the concept that dispersion of compounds in the liver is primarily determined by the heterogeneity of the hepatic microvasculature. The relationship between PS values in the perfused rat liver (either abstracted or taken from the literature data) and physicochemical properties for 17 compounds has been explored. There appears to be a continuous relationship between PS and logD, a measure of lipophilicity that takes into account the degree of ionization in the perfusate. The PS value for EBA is close to that expected based on its physicochemical properties. PMID- 7473186 TI - Pretreatment with a blocking monoclonal antibody to P-selectin accelerates pharmacological thrombolysis in a primate model of arterial thrombosis. AB - It has been established that the fibrin content of a developing thrombus can be dramatically reduced with the use of the GA6 monoclonal antibody, which is directed against P-selectin (CD62p). This effect is probably related to diminished tissue factor activity on monocytes in the presence of P-selectin antagonism. Therefore, we hypothesized that an occlusive arterial thrombus formed in the presence of a P-selectin monoclonal antibody would be more susceptible to lysis with standard thrombolytic therapy. To test this hypothesis, 22 male cynomolgus monkeys were anesthetized and instrumented for induction of thrombosis of a femoral artery. Endothelial injury was induced by passing a 150-microA anodal current through a small electrode that was placed in the femoral artery. Blood flow through the artery was continuously monitored using an ultrasonic transit-time flowmeter. The GA6 monoclonal antibody (1 mg/kg) or control, isotype matched mouse IgG1 (P23 or P7) was administered i.v. 1 hr before electrolytic endothelial injury. In the P23 group (n = 11), an occlusive thrombus formed in 52.1 +/- 8.5 min, and in the GA6 group (n = 11), an occlusive thrombus formed in an average time of 52.0 +/- 8.1 min. After formation of an occlusive thrombus, the current was terminated and intravenous heparin (100 U/kg + 50 U/kg/hr) was administered to prevent clot extension.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7473187 TI - Mechanisms of ethanol-induced suppression of a primary antibody response in a mouse model for binge drinking. AB - A previous study in this laboratory demonstrated that a single dose of EtOH causes thymic atrophy and that elevated levels of endogenous glucocorticoids are responsible, at least in part, for this effect. In the present study using the same animal model, the effect of EtOH on the T-dependent antibody response to sheep red blood cells (SRBC) was evaluated. In addition, the effects of EtOH on the SRBC-induced expression of selected cytokines known to enhance or regulate humoral responses were evaluated. Administration of EtOH at 5.0 to 7.0 g/kg to B6C3F1 mice significantly and dose-responsively inhibited the generation of antibody-forming cells (AFC) after immunization in vivo or in vitro in Mishell Dutton cultures. In contrast, direct addition of EtOH or its major metabolites to Mishell-Dutton cultures did not decrease the AFC response. In vivo administration of EtOH caused substantial decreases in the SRBC-induced expression of interleukin (IL)-1-beta, IL-2 and IL-4 mRNA in the spleen, as indicated by reverse transcription-polymerase chain reaction. The suppressed AFC responses and suppressed cytokine gene expression were prevented by RU 486, a potent glucocorticoid antagonist. Thus, suppression of cytokine gene expression by elevated levels of endogenous glucocorticoids may play a role in EtOH-induced suppression of the primary humoral response to SRBC. However, differences in dose response patterns for suppression of cytokine gene expression and suppression of the AFC response suggest the involvement of additional factors. PMID- 7473188 TI - Mechanisms of absorption enhancement by medium chain fatty acids in intestinal epithelial Caco-2 cell monolayers. AB - Sodium salts of medium chain fatty acids (MCFAs) enhance the absorption of hydrophilic drugs across the intestinal mucosa, but the mechanism behind the effect is largely unknown. In this study, the dose-dependent effects of the sodium salts of four MCFAs, C6 (caproate), C8 (caprylate), C10 (caprate) and C12 (laurate), on the permeability of the hydrophilic marker molecule [14C]mannitol were studied in monolayers of the human intestinal epithelial cell line, Caco-2, grown on permeable supports. C8, C10 and C12, but not C6, enhanced the permeability of [14C]mannitol in a dose-dependent manner. Comparison of the cellular effects of the MCFAs at concentrations that gave comparable (8.1- to 8.5 fold) absorption enhancement showed that: 1) C8 was active as absorption enhancer only when the tonicity of the medium was increased; 2) absorption enhancement mediated by C10 was related to a redistribution of the cytoskeleton and structural dilatations in the tight junctions; and 3) C12 was without effect on the cytoskeleton and cellular morphology. Studies on C10 under anisotonic conditions showed that deviations from isotonicity enhanced its effect. These results suggest that structurally similar MCFAs display dramatic differences in their mechanism of action. In addition, the effects of osmolality provide an explanation for the previously reported variability in the efficacy of MCFAs as absorption enhancers. PMID- 7473189 TI - Tissue distribution of cocaine methyl esterase and ethyl transferase activities: correlation with carboxylesterase protein. AB - The tissue distribution of cocaine methyl esterase and ethanol-dependent ethyl transferase activities was determined in the rat and compared to the tissue distribution of three distinct non-specific hydrolases. Rates of formation of benzoylecgonine from cocaine and cocaethylene from ethanol and cocaine were measured in serum and tissue homogenate-supernatants of the brain, heart, kidney, liver, lung and spleen. The tissue distribution of three nonspecific esterases, A, B and C, was defined by nondenaturing gel electrophoresis and measuring the hydrolysis of 4-methylumbelliferyl acetate in the gels. Immunoreactive protein was localized by using Western blot analysis with polyclonal rabbit antihuman liver cocaine methyl esterase antibody after denaturing and nondenaturing gel electrophoresis. The rat liver, lung, kidney and heart exhibited cocaine methyl esterase and ethyl transferase activities and immunoreactive protein. The brain had cocaine methyl esterase activity but no ethyl transferase activity; neither activity was found in serum or spleen. The dominant immunoreactive bands in the liver, lung, kidney and heart comigrated with the 59 kD band of purified human liver cocaine methyl esterase. The rat liver, lung and kidney exhibited a band of nonspecific esterase activity that migrated with purified human liver cocaine methyl esterase and rat hydrolase A. These observations suggest that rat hydrolase A is similar to human cocaine methyl esterase. The lack of straight forward correlation between cocaine methyl esterase activity and immunoreactive protein and nonspecific esterase activity suggests that more than one enzyme catalyzes the hydrolysis of cocaine to benzoylecgonine in the rat. PMID- 7473190 TI - Age- and sex-related alterations of microsomal drug- and testosterone-oxidizing cytochrome P450 in Sprague-Dawley strain-derived dwarf rats. AB - Effect of growth hormone (GH) on the age-related changes in hepatic cytochrome P450 (P450) was studied using GH-deficient dwarf and parental Sprague-Dawley rats. Microsomal testosterone (T) T2 alpha- and T2 beta-hydroxylations were lower in livers of mature male dwarf rats than the normals, whereas T16 beta hydroxylation was rather higher in male dwarf rats. Although T2 alpha-, T2 beta-, T6 beta-, T16 alpha- and T16 beta-hydroxylations were barely detectable in senescence normal rats (24 months old), considerable levels of T6 beta-, T16 alpha- and T16 beta-hydroxylations were maintained in senescence dwarf rats (after 22 months old). These results are caused by the alteration of specific P450 forms including CYP2B1, CYP2B2, CYP2C11 and CYP3A2 in dwarf rats. Appearance of male-specific CYP2C11 and CYP3A2 and high levels of CYP2B1 and CYP2B2 in female dwarf rats indicate the role of pituitary GH on liver of normal rats. However, the additional role of a factor other than GH was suggested on the sex related differences and age-associated alterations of specific P450 contents in dwarf rats. CYP2C11 appears in dwarf female rats with the same developmental profile as observed in normal male rats. This form appears apparently with the development of GH receptor in livers, suggesting the possibility that a factor independent from androgen and GH governs the ontogeny of this P450 in the liver. A female-specific protein, CYP2C12, in normal rat livers, also appeared in both sexes of senescence dwarf rats, suggesting the role of non-GH factor on the expression of this P450 in liver. PMID- 7473191 TI - Characterization of alpha 2 adrenoceptors and other adrenoceptors in membranes isolated from dog mesenteric nerve axons. AB - The axonal membranes from dog mesenteric nerves were isolated and characterized. They had a high density of [3H]saxitoxin, [3H]rauwolscine and [125I]iodocyanopindolol binding sites, but low densities of [125I] omega conotoxin (GVIA) and [3H]prazosin binding sites. The microsomal membranes had one of the higher densities of [3H]rauwolscine binding sites in centrifugation fractions. These sites were characterized by saturation, kinetic and competition studies. In competition with oxymetazoline, WB 4101, prazosin, ARC-239, SK&F 104078 and SK&F 104856, these rauwolscine binding sites were alpha 2A-like. Chloroethylclonidine pretreatment did not affect the ability of SK&F 104078 or SK&F 104856 to compete with rauwolscine binding sites on axonal membranes. Bmax values were unchanged by chloroethylclonidine under conditions that also failed to reduce Bmax values of rauwolscine binding to membranes from human platelets. These receptors appear to be alpha 2A adrenoceptors and may represent sites of prejunctional control of neurotransmitter release in the dog. This membrane preparation may be useful in biochemical studies of prejunctional neural receptors. PMID- 7473193 TI - Anti-inflammatory properties of Go 6850: a selective inhibitor of protein kinase C. AB - Protein kinase C (PKC) regulates a variety of signal transduction events implicated in the pathogenesis of inflammation, including the biosynthesis of inflammatory cytokines and superoxide and the activation of phospholipase A2. Because of the significant role of PKC in these inflammatory processes, we evaluated a specific and potent inhibitor of C kinase for efficacy in several in vitro and in vivo murine models of inflammation. Unlike the relatively nonspecific kinase inhibitor staurosporine, the bisindolylmaleimide 3-[1-[-3 (dimethylaminopropyl]-1H-indol-3-yl]- 4-(1H-indol-3-yl)-1H-pyrrole-2,5-dione monohydrochloride (Go 6850) demonstrated increased selectivity for C kinase in purified enzyme assays (respective IC50 values (microM) for Go 6850 and staurosporine: protein kinase C (0.032, 0.009); myosin light-chain kinase (0.6, 0.01); protein kinase G (4.6, 0.018); protein kinase A (33, 0.04); tyrosine kinase1 (94, 0.4); tyrosine kinase2 (> 100, > 1)). Topically applied Go 6850 inhibited phorbol myristate acetate-induced edema, neutrophil influx and vascular permeability in murine epidermis in a dose- and time-dependent manner at levels comparable to indomethacin. In a murine model of delayed type hypersensitivity, Go 6850 inhibited dinitrofluorobenzene-induced contact dermatitis with and ID50 value of 150 micrograms/ear. Cellular studies in mouse peritoneal macrophages demonstrated that Go 6850 was a potent inhibitor of phorbol myristate acetate induced prostaglandin E2 production. Superoxide production in phorbol myristate acetate-stimulated murine neutrophils was also inhibited by Go 6850 (IC50 = 88 nM).(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7473192 TI - Mechanisms of up-regulation of neuronal nicotinic acetylcholine receptors in clonal cell lines and primary cultures of fetal rat brain. AB - There is a consensus that high-affinity [3H]-L-nicotine binding sites in the mammalian brain, which are thought to represent a predominant form of central nervous system nicotinic acetylcholine receptor (nAChR) composed of alpha 4 and beta 2 subunits, are increased in number after chronic nicotine exposure. However, mechanisms responsible for this effect have not yet been elucidated. To evaluate this issue, we have used, as models, primary cell cultures of fetal rat brain cortex, in which high-affinity [3H]-L-nicotine binding sites are naturally expressed, and clonal cell cultures of fibroblasts stably transfected to express nAChR composed of transgenic chick alpha 4 and beta 2 subunits under control of dexamethasone-inducible promoters. Chronic nicotine exposure induced an approximately 2.5-fold increase in high-affinity [3H]-L-nicotine binding sites in M10 cells maintained in the presence of dexamethasone or in primary fetal rat brain cortical cultures. Up-regulation of [3H]-L-nicotine binding sites was evident for M10 cells treated at nicotine concentrations as low as 10 nM (EC50 and EC100 values were 100 nM and 10 microM, respectively). Scatchard analyses of [3H]-L-nicotine binding data in M10 cells indicated a change in Bmax with no significant change in affinity for radioligand (KD = 2.5 +/- 0.5 nM in control cells vs. 2.0 +/- 0.4 nM in nicotine-treated cells). Northern blot analyses indicated that nicotine treatment alone had no direct effect on the promoter driving transgenic nAChR subunit gene transcription in M10 cells and that steady state levels of fetal rat brain cortical cell or M10 cell nAChR alpha 4 or beta 2 mRNAs were unaffected under conditions of chronic nicotine treatment that produced up-regulation of high-affinity [3H]-L-nicotine binding sites.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7473194 TI - Influence of permeating ions on potassium channel block by external tetraethylammonium. AB - 1. Block by external tetraethylammonium (TEA) was examined on currents carried by potassium (K+) and sodium (Na+) through the cloned delayed rectifier K+ channel Kv2.1. 2. In the presence of high intracellular K+ and high extracellular Na+, currents through the Kv2.1 channel were carried almost exclusively by K+. In the absence of both intracellular and extracellular K+, large currents were carried by Na+ through the Kv2.1 channel. 3. External application of 30 mM TEA blocked K+ currents through Kv2.1 by 87%. The same external concentration of TEA had no effect on Na+ currents through this channel. 4. As the K+ concentration was increased between 0 and 140 mM, the percentage of current blocked by TEA progressively increased from 0 to 87%. 5. These data indicate that block of K+ channels by external TEA depends on the permeating ion. Furthermore, these data indicate that in the presence of Na+ and absence of K+, TEA does not bind to the channel, and that addition of low concentrations of K+ facilitates TEA binding. PMID- 7473195 TI - Histamine and ATP mobilize calcium by activation of H1 and P2u receptors in human lens epithelial cells. AB - 1. Cytoplasmic calcium concentration ([Ca2+]1) of single superfused tissue cultured human lens epithelial cells (HLEC) was monitored using the fluorescent dye fura-2; the resting values were low and stable for several hours ([Ca2+]i = 96 +/- 20 nM; mean +/- S.D., n = 16). 2. Continuous superfusion with either ATP or histamine (0.1-10 microM) produced regular oscillations in [Ca2+]i that could be maintained for a short time in the absence of external calcium. 3. Short (30 s) pulses of histamine (0.1-100 microM) induced a transient rise in [Ca2+]i, the time course of which was insensitive to the removal of external calcium. The rate of rise and the amplitude of the response were very sensitive to agonist concentration, whereas the rate of recovery was relatively constant. 4. The responses to long pulses of histamine (> 100 s) consisted of an initial transient followed by a maintained [Ca2+]i which returned to baseline on removal of external calcium. 5. The kinetics of the responses to short and long pulses of ATP (0.1-100 microM) were very similar to those of histamine and showed a similar sensitivity to the presence or absence of external calcium. 6. The histamine responses were abolished by triprolidine (1 microM), but unaffected by ranitidine (1 microM), indicating that an Hi receptor subtype is activated by histamine. 7. The ATP responses were reversibly inhibited by suramin and the potency sequence for a range of agonists was ATP = UTP = ATP gamma S > ADP = GTP >> AMP = adenosine, indicating that activation of a P2u receptor subtype was responsible for the increase in [Ca2+]i. 8. Both histamine and ATP responses were abolished by thapsigargin (100 nM), confirming that calcium release from intracellular stores was responsible for the initial peak of the response. Application of either agonist during the plateau phase of the thapsigargin response often led to a marked, but reversible, decline in [Ca2+]i, indicating the presence of a further, normally hidden, calcium regulatory factor associated with the presence of the agonist. 9. Maximal concentrations of either histamine or ATP totally emptied the calcium store as a subsequent application of the other agonist (or thapsigargin), in the absence of external calcium, failed to induce a further increase in the calcium signal. PMID- 7473196 TI - Imaging of calcium variations in living dendritic spines of cultured rat hippocampal neurons. AB - 1. Cultured rat hippocampal neurons were loaded with the Ca2+ indicator fura-2 through micropipettes and visualized with an inverted microscope equipped with a high power objective and a cooled CCD camera. The responses of dendritic spines and their parent dendrites to stimuli which evoke a rise of [Ca2+]i were monitored. 2. NMDA caused a rapid and transient rise in [Ca2+]i, which was more evident in the spine than in the parent dendrite. The recovery in both compartments had the same time course, and was dependent on normal [Na+]o. 3. Application of alpha-latrotoxin, which causes release of neurotransmitters from terminals, produced a rise of [Ca2+]i in the dendritic spines, more than in their parent dendrites. Prolonged exposure to the drug eliminated the spine/dendrite disparity. 4. The presence of voltage-gated calcium channels in dendritic spines is indicated by the enhanced calcium rise in spines rather than dendrites of cells depolarized by either intracellular current injection or by raising [K+]o. This rise was attenuated by nifedipine or verapamil, both L-type channel blockers. 5. It is suggested that the dendritic spine constitutes an independent calcium compartment that is closely linked to the parent dendrite. PMID- 7473197 TI - Intracellular spermine confers rectification on rat calcium-permeable AMPA and kainate receptors. AB - 1. Whole-cell recordings were made from cerebellar granule cells cultured in high K+ medium to induce expression of Ca(2+)-permeable AMPA receptors. Current voltage (I-V) plots of agonist-evoked responses showed varying degrees of inward rectification, but became linear within 5-10 min. 2. Recombinant Ca(2+)-permeable kainate receptors, composed of GluR6(Q)/KA-2 subunits, exhibited rectifying whole cell I-V plots that became linear in outside-out patches. 3. Loss of rectification in granule cells was prevented by including 100 microM spermine in the pipette; the degree of rectification was then correlated with Ca2+ permeability. 4. Spermine also prevented loss of rectification in patches containing GluR6(Q)/KA-2 receptors (IC50, 1.7 microM). 5. We suggest that spermine, or a similar cellular constituent, may act as a cytoplasmic factor conferring inward rectification on Ca(2+)-permeable non-NMDA receptors, and that 'washout' of this factor underlies the observed loss of rectification. PMID- 7473198 TI - Block of native Ca(2+)-permeable AMPA receptors in rat brain by intracellular polyamines generates double rectification. AB - 1. The influence of intracellular factors on current rectification of different subtypes of native alpha-amino-3-hydroxy-5-methyl-4-isoxazoleproprionate receptors (AMPARs) was studied in rat brain slices by combining fast application of glutamate with patch pipette perfusion. 2. The peak current-voltage (I-V) relation of the AMPARs expressed in Bergmann glial cells of cerebellum and dentate gyrus (DG) basket cells of hippocampus was weakly rectifying in outside out patches and nystatin-perforated vesicles, but showed a doubly rectifying shape with a region of reduced slope between 0 and +40 mV in nucleated patches. The I-V relation of AMPARs expressed in hippocampal CA3 pyramidal neurones was linear in all recording configurations. 3. Intracellular application of 25 microM spermine, a naturally occurring polyamine, blocked outward currents in outside out patches from Bergmann glial cells and DG basket cells in a voltage-dependent manner, generating I-V relations with a doubly rectifying shape which were similar to those recorded in nucleated patches. AMPARs in CA3 pyramidal cell patches were unaffected by 25 microM spermine. 4. The half-maximal blocking concentration of spermine at +40 mV was 0.3 microM in Bergmann glial cell patches and 1.5 microM in DG basket cell patches, whereas it was much higher (>> 100 microM) for CA3 pyramidal cell patches. Spermidine also affected current rectification, but with lower affinity. The block of outward current by polyamines following voltage jumps developed within < 0.5 ms. 5. We conclude that current rectification, rather than being an intrinsic property of the Ca(2+) permeable AMPAR channel, is generated by polyamine block. PMID- 7473199 TI - Voltage-dependent calcium currents are enhanced in dorsal root ganglion neurones from the Bio Bred/Worchester diabetic rat. AB - 1. Whole-cell, high-threshold, voltage-dependent calcium currents (ICa) were enhanced in acutely dissociated, capsaicin-sensitive dorsal root ganglion neurones from diabetic Bio Bred/Worchester (BB/W) rats, compared with those from age-matched, non-diabetic controls. The magnitude of the enhancement increased with the duration of diabetes, and reached significance at diabetic durations of 6 months (diabetic: 6.3 +/- 0.4 nA; current density (CD), 157 +/- 12 pA pF-1; means +/- S.E.M., n = 9, P < 0.01; control: 3.9 +/- 0.6 nA; CD, 116 +/- 11 pA pF 1; n = 18) and 8 months (diabetic: 7.6 +/- 0.4 nA; CD, 177 +/- 25 pA pF-1; n = 11, P < 0.005; control: 5.1 +/- 0.5 nA; CD, 111 +/- 26 pA pF-1; n = 15). Low threshold, voltage-dependent ICa were also enhanced in neurones from animals diabetic for 8 months (diabetic: 2.5 +/- 0.7 nA, n = 4, P < 0.05; control: 0.7 +/ 0.5 nA, n = 6). 2. The ICa enhancement was prevented by long-term treatment of diabetic animals with an aldose reductase inhibitor (ARI; peak ICa at 6 months: 4.41 +/- 0.48 nA, n = 2; at 8 months: 4.32 +/- 0.60 nA, n = 9). 3. The ICa enhancement was not due to a shift in the voltage dependence of either the current-voltage relationship or steady-state inactivation. 4. The L channel antagonist nifedipine and preferential N channel antagonist omega-conotoxin GVIA (omega-CgTX) caused a greater inhibition of high-threshold ICa in diabetic neurones compared with controls (nifedipine: control: 25 +/- 3%, n = 26; diabetic: 36 +/- 7%, n = 11; omega-CgTX: control: 40 +/- 4%, n = 21; diabetic: 50 +/- 7%, n = 7). Diabetic neurones also demonstrated a significantly greater residual current (2.44 +/- 0.34 nA, n = 7) in the presence of both antagonists vs. controls (1.28 +/- 0.30 nA, n = 8, P < 0.05), suggesting that N-, L- and additional non-N-, non-L-type high-threshold ICa were enhanced. PMID- 7473200 TI - ATP-activated inward current and calcium-permeable channels in rat macrophage plasma membranes. AB - 1. To study mechanisms of receptor-operated Ca2+ influx in non-excitable cells, membrane currents of rat peritoneal macrophages were recorded using whole-cell cell-attached and outside-out configurations of the patch clamp technique. Under whole-cell recording conditions, ATP applied in micromolar concentrations elicited an inward current response when the bath solution contained Ba2+, Ca2+ or Na+ as the only permeant cations. 2. Increasing the Mg2+ concentration had an inhibitory effect on the ATP-induced inward current indicating that the active form of ATP responsible for the cation entry is ATP4-. The nucleotide potency order was ATP > ATP gamma S > ADP. UTP was completely ineffective (n = 19). The data obtained are consistent with the ATP receptor being of the P2Z type. 3. The macrophage plasma membrane was impermeable to Tris+ during the ATP-induced current at ATP4- concentrations varying from 0.07 to 500 microM. At higher concentrations, ATP produced a large inward steady-state current, which could be attributed to membrane permeabilization. 4. Activity of single channels was recorded when ATP was applied to the external surface of the patch membrane both in cell-attached and outside-out experiments. A specific property of the channels appeared to be the existence of at least four conductance sublevels. With 105 mM Ba2+ as the permeant cation, the conductance sublevels were 3.5, 7, 10 and 15 pS. With 10 mM Ca2+ the sublevel conductances were equal to 4, 9, 13 and 17 pS. 5. The unitary conductance estimated from the whole-cell current noise analysis (3.5 4.5 pS for 105 mM Ba2+) was significantly lower than that obtained from single channel measurements at the main (3rd) current level, but it was very close to the conductance of the minimum (1st) level. 6. Extrapolated reversal potential values estimated from current-voltage curves for predominant conductance levels were equal to +40 and +26 mV for 105 mM Ba2+ and 10 mM Ca2+, respectively. The permeability ratios fell in the sequence: PCa:PBa:PK = 71.:29:1. Thus, ATP activated channels in the macrophage membrane are rather selective for divalent vs. monovalent cations, with the predominant permeability being for Ca2+. PMID- 7473201 TI - ATP-operated calcium-permeable channels activated via a guanine nucleotide dependent mechanism in rat macrophages. AB - 1. To elucidate the possible involvement of a G protein in ATP-evoked Ca(2+) permeable channel activity, membrane currents of rat peritoneal macrophages were recorded using inside-out and cell-attached configurations of the patch clamp technique. 2. In inside-out experiments with a pipette solution containing 105 mM Ba2+, application of 100 microM GTP or GTP gamma S to the internal surface of the membrane elicited a rise in channel activity. This effect was observed in 49% of the patches investigated (n = 69). The mean value of NPo (N, number of open channels; Po, channel open probability) was equal to 0.49 +/- 0.27 (mean +/- S.E.M.; n = 16). The delay in the activity development was 21 +/- 8 s (n = 18) with 200 microM ATP added to the pipette solution and about 4 min (n = 5) without agonist in the pipette. Similar results were obtained with 10 mM Ca2+ as the only permeant cation. 3. Properties of GTP gamma S-evoked channels were identical to those of channels activated by extracellular application of ATP. The channels exhibited at least four conductance sublevels, the 4th one being the least frequent. With 105 mM Ba2+ as a permeant cation, sublevel conductances were 3.5, 7, 10 and 15 pS. Corresponding values for 10 mM Ca2+ were about 4, 9, 13 and 17 pS. Extrapolated reversal potential (Er) values were about +40 and +25 mV for Ba2+ and Ca2+, respectively. 4. The activity of channels with similar characteristics could be induced by the extracellular application of fluoride in cell-attached experiments without any agonist in the pipette solution.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7473202 TI - Control of Ca2+ entry into rat lactotrophs by thyrotrophin-releasing hormone. AB - 1. Lactotrophs are adenohypophysial cells that synthesize and secrete prolactin (PRL), a hormone principally involved in mammalian milk production. An increase in the intracellular Ca2+ concentration ([Ca2+]i) is an important signal for PRL secretion. Thyrotrophin-releasing hormone (TRH) generates Ca2+ signals derived from both the release of Ca2+ from intracellular stores and the entry of extracellular Ca2+, the latter being particularly important for PRL secretion. The identity of this TRH-sensitive Ca2+ entry pathway is unknown and therefore the subject of the present study. 2. [Ca2+]i was measured by video imaging of fura-2 loaded into single rat anterior pituitary cells. Ca2+ influx was detected by quenching of fura-2 fluorescence by external Mn2+. All data are from lactotrophs isolated from lactating female rats. Individual lactotrophs were identified by postexperimental immunofluorescent detection of PRL in fixed cells. 3. TRH induced the release of Ca2+ from intracellular stores and also stimulated Mn(2+)-permeable Ca2+ influx. U73122 (1 microM), a phospholipase C inhibitor, prevented the Ca(2+)-mobilizing actions of TRH. The chemically similar but inactive analogue, U73343 (1 microM), had no effect on TRH responses. U73122 did not act as a global G protein inhibitor because the reduction of basal [Ca2+]i by dopamine (1 microM, a G protein-mediated event) was not affected. 4. TRH stimulated Mn2+ influx occurred either immediately after addition of TRH (early entry) or after a delay of about 130 s (late entry). There were no statistically significant differences in the magnitude or temporal characteristics of the Ca2+ signals evoked from cells showing early or late Mn2+ entry. 5. The identity of Ca2+ channels permeable to Mn2+ was investigated. Cell depolarization with 50 mM KCl stimulated Ca2+/Mn2+ influx and was prevented by nifedipine (1 microM). Bay K 8644 (1 microM) also stimulated Mn2+ influx. Thus, the presence of Mn(2+) permeable L-type voltage-operated Ca2+ channels is likely. A second Mn(2+) permeable pathway was present in lactotrophs. Depletion of Ca2+ stores by thapsigargin (1 microM) stimulated a Ca2+ signal and Mn2+ influx. This 'capacitative entry pathway' was insensitive to nifedipine (1 microM), indicating that putative L-type Ca2+ channels were not activated. 6. TRH-stimulated Mn2+ influx was not prevented by nifedipine (1 microM). TRH added during KCl-induced Mn2+ influx reduced the quench rate within the time frame of the TRH-induced Ca2+ spike. TRH may therefore inhibit putative L-type Ca2+ channels. 7. Addition of thapsigargin in Ca(2+)-free medium transiently increased [Ca2+]i and prevented subsequent Ca2+ responses to TRH.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 7473203 TI - Amino acid-induced [Ca2+]i oscillations in single mouse pancreatic islets of Langerhans. AB - 1. The effects of amino acids on cytosolic free calcium concentration ([Ca2+]i) were measured, using fura-2 fluorescence imaging, in mouse pancreatic islets of Langerhans. 2. Slow [Ca2+]i oscillations appeared when isolated islets were incubated with a solution containing a mixture of amino acids and glucose at concentrations found in the plasma of fed animals. 3. In the presence of 11 mM glucose, alanine (5 mM) and arginine (10 mM) induced a transient rise in [Ca2+]i followed by an oscillatory pattern, while leucine (3 mM) and isoleucine (10 mM) triggered the appearance of slow [Ca2+]i oscillations. 4. Also in the presence of glucose (11 mM), tolbutamide (10 microM) increased the duration of the glucose induced [Ca2+]i oscillations. While tolbutamide (10 microM) did not modify the leucine-induced slow oscillatory pattern, addition of diazoxide (10 microM) resulted in the gradual appearance of [Ca2+]i oscillations which resembled the glucose-induced fast oscillations. 5. Like stimulatory glucose concentrations (11 mM), glyceraldehyde (10 mM) induced fast oscillations of [Ca2+]i. 6. Fluoroacetate (2 mM) transformed leucine-induced slow [Ca2]i oscillations into fast [Ca2+]i oscillations. Iodoacetate (1 mM) completely inhibited any oscillatory pattern. 7. It is suggested that mitochondrially generated signals, derived from amino acid oxidative metabolism, acting in conjunction with glucose signalled messengers, are very effective at closing ATP-dependent K+ channels (KATP+). 8. We propose that metabolic regulation of KATP+ channels is one of the mechanisms underlying the modulation of the oscillatory [Ca2+]i response to nutrient secretagogues. PMID- 7473204 TI - Stretch-induced enhancement of contractions in uterine smooth muscle of rats. AB - 1. We studied the effect of servo-controlled stretch of smooth muscle strips from rat uterus on tension and intracellular Ca2+ concentration ([Ca2+]i, using fura-2 as an indicator) at 30 degrees C. 2. When quiescent uterine muscle strips were stretched at a ramp time of 0.5 s by multiples of 5% of the resting muscle length (L0) up to 40%, forty-two out of sixty muscle strips responded with a transient active contraction and a [Ca2+]i increase. The minimum excursion of stretch required for contraction was 26.3 +/- 7.5% of L0 (mean +/- S.D.). The peak response had an all-or-none property and was almost independent of the duration of stretch. 3. Stretches of 30 or 35% of L0 induced contraction in most cases when rapidly applied in 0.2-0.5 s, but slowly applied stretch (ramp duration of 5 10 s) rarely induced contraction. 4. The stretch-induced response was inhibited by the removal of extracellular Ca2+ or by the addition of 10 nM nicardipine. However, it was unaffected by 1 microM tetrodotoxin, 1 microM atropine or by 10 microM cyclopiazonic acid, an inhibitor of Ca2+-ATPase in intracellular Ca2+ stores. 5. When a stretch of 15-35% of L0 was applied during the relaxation phase of 10 nM oxytocin-induced rhythmic contractions, the first contraction after the stretch occurred earlier than that expected from the control rhythm. However, the frequency of the subsequent rhythm returned to almost the control level even during continued application of stretch, although the half-width of rhythmic contractions was increased during stretch. 6. The present study demonstrates that stretch of uterine muscle induces a transient contraction due to Ca2+ influx, which is myogenic and dependent on the excursion and velocity of stretch. The all or-none property of the stretch-induced contractions suggests initiation of Ca2+ spikes. Furthermore, stretch modulates the oxytocin-induced rhythmic contractions. PMID- 7473205 TI - Properties and roles of an intramembranous charge mobilized at high voltages in frog skeletal muscle. AB - 1. Membrane Ca2+ currents (ICa), intramembranous charge movement currents and changes in intracellular Ca2+ concentrations were recorded in voltage clamped cut skeletal muscle fibres of the frog. Intra- and extracellular solutions, designed to prevent ionic current, and use of the saponin-permeabilization procedure made possible the measurement of transfer of intramembranous charge up to high positive potentials. 2. Substantial charge moved at positive potentials. This charge was shown to be intramembranous in four tests of charge conservation, demonstrating that the total displacement of charge depended only on the initial and final voltages, and not on the history or pathway of intermediate voltages. 3. On average, in twenty-three cells, the charge moved at 50 mV was 31 +/- 1.9 nC microF-1 (mean +/- S.E.M.), and at 0 mV was 25 +/- 1.5 nC microF-1. Approximately one-fifth of the total charge moved above 0 mV. 4. The charge that moved at high voltage could be fitted, in most cases, with a Boltzmann distribution function. In twenty of twenty-three cells, the total charge distribution could be fitted as the sum of two Boltzmann terms; the high voltage term was centred at 11 +/- 3.9 mV, with a steepness factor of 12 +/- 1.6 mV and a magnitude of 8.6 +/- 1.1 nC microF-1. The low voltage term was centered at -43 +/- 2.1 mV, with a steepness factor of 7.7 +/- 0.6 mV and a magnitude of 22 +/- 1.8 nC microF-1. Thus, the high voltage component comprised about one-quarter of the mobile charge. In four cells it was possible to fit the sum of three Boltzmann terms to the distribution of mobile charge; the parameters of the high voltage term then were similar to those found by fitting the sum of two Boltzmann terms to the same data. 5. The voltage dependence of activation of ICa was determined in a buffered 2 mM Ca2+ external solution, from the tails of ionic current at -30 mV, after activating pulses to various voltages, the duration of which was sufficient to reach the peak of inward current. The voltage dependence was described by a Boltzmann function centred at 2.6 +/- 6.9 mV (n = 6), with a steepness factor of 20 +/- 1.4 mV. The voltages at which the high voltage charge moved were roughly the same as those at which ICa was activated. 6. Calcium release from the sarcoplasmic reticulum was determined from the Ca2+ transients. Calcium release continued to increase at potentials above 0 mV.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 7473206 TI - Action potential generation in rat slow- and fast-twitch muscles. AB - 1. In skeletal muscle fibres, voltage-gated sodium channels are concentrated at the neuromuscular junction. The effect of this accumulation of sodium channels on action potential generation was investigated in rat slow- and fast-twitch muscle fibres. 2. Intracellular microelectrodes were used to generate and record action potentials, from an imposed membrane potential of -75 and -90 mV, in junctional and extrajunctional regions of the muscle fibre. To identify junctional regions, preparations were incubated with 5 x 10(-7) M d-tubocurarine (dTC) to block muscle contraction in response to nerve stimulation whilst allowing endplate potentials (EPPs) to be recorded. Injection of rectangular depolarizing current pulses initiated action potentials at the endplate with threshold values several millivolts lower than those generated elsewhere in the fibre. In addition, the maximum rate of rise of the action potential was greater at the endplate than in extrajunctional regions. 3. In other muscles, neuromuscular transmission was partially blocked with dTC (2 x 10(-7) M), such that repetitive nerve stimulation evoked action potentials and EPPs in the same fibre. The threshold of these nerve evoked action potentials was approximately 50% lower than values derived from action potentials generated by current injection. 4. It is concluded that the threshold for action potential generation is significantly lower at the neuromuscular junction than in extrajunctional regions of skeletal muscle fibres. Furthermore, nerve-evoked current is more effective at generating an action potential than is injected current. PMID- 7473207 TI - Burst kinetics of single NMDA receptor currents in cell-attached patches from rat brain cortical neurons in culture. AB - 1. The patch-clamp technique was used to record single-channel currents from cell attached patches on rat brain cortical neurons in culture. The composition of the open and shut intervals during bursts of openings was studied in N-methyl-D aspartate (NMDA) receptors exposed to 1 microM NMDA and 10 microM glycine at a membrane potential of -70 mV. 2. Open interval histograms were constructed for openings at each position (first, second, third, etc.) during all bursts. Distributions from openings two to five were fitted with similar (two or three) exponential components. The first opening from all bursts tended to be of shorter duration than the other openings. 3. Bursts were sorted according to the number of openings they contained and duration histograms were obtained for the openings at each position (one to five) during bursts of up to five openings. For bursts containing two or more openings, the distribution of open durations obtained at a given position were similar to each other regardless of the number of openings in the burst. 4. In bursts of two or more openings (up to five), duration histograms from the openings at each position in the burst were fitted with two or three exponential components that were similar for each opening. Bursts consisting of a single opening had a different distribution, having a relatively larger component of short duration. 5. Shut intervals during bursts were described by two exponential components with average time constants (and relative areas) (means +/ S.E.M.) of 0.06 +/- 0.01 ms (0.59 +/- 0.02) and 0.64 +/- 0.02 ms (0.41 +/- 0.02). Their distribution was independent of the numbers of openings in the bursts, their position within the burst, and the types of openings (long or short duration) contained within the burst. 6. These results suggest that each opening in bursts of two or more openings is kinetically similar to every other opening regardless of burst length. Analogously, each shut period during a burst was similar to every other. A kinetic model with three open and four closed intraburst states is shown to be consistent with these results for bursts of two or more openings. PMID- 7473208 TI - Contribution of SK and BK channels in the control of catecholamine release by electrical stimulation of the cat adrenal gland. AB - 1. Transmural electrical stimulation (10 Hz, 1 ms, 40 V for 10 s) of cat adrenal glands perfused at room temperature with Krebs-Hepes solution produced catecholamine secretory responses which were reproducible when stimulations were applied at 5 min intervals. Such responses were inhibited about 20% by atropine (1 microM) and 80% by hexamethonium (30 microM). Apamin (100 nM) increased the secretory response 2.5-fold in the presence of atropine and 8-fold in the presence of hexamethonium. 2. Potentiation by apamin of secretory responses evoked by 100-pulse trains was similar at 5, 10 and 20 Hz (about 2-fold). When glands were continuously stimulated at 3 Hz, apamin increased 4-fold the initial secretion plateau. Continuous stimulation at a higher frequency (20 Hz) produced a sharp secretory peak followed by a small, sustained plateau; apamin did not alter this plateau. Apamin also enhanced the secretory responses obtained with sustained stimulation with acetylcholine (10 or 200 microM). 3. Secretion peaks induced by brief acetylcholine pulses (10 microM for 10 s) applied to isolated and superfused cat adrenal chromaffin cells were enhanced more than 3-fold by 100 nM apamin. Charybdotoxin (10 nM) did not enhance these secretory peaks. 4. In perfused cat adrenal glands, charybdotoxin (10 nM) affected neither the secretion evoked by trains of electrical stimulation applied at different frequencies nor the secretion evoked by acetylcholine pulses. 5. In 0.5 mM [Ca2+]o, apamin enhanced 3-fold the secretion evoked by electrical stimulation trains of 100 pulses (10 Hz, 10 s) and almost 6-fold the acetylcholine (10 microM for 10 s) induced secretion. In 5 mM Ca2+, apamin enhanced the secretory responses to electrical stimulation and acetylcholine 2- and 10-fold, respectively. Charybdotoxin enhanced 2.5-fold the secretory response to electrical stimulation in 0.5 mM Ca2+, although this effect was not statistically significant. A synergistic interaction between the two toxins on catecholamine release induced by electrical stimulation was observed at low but not at high [Ca2+]o. 6. Simultaneous release of acetylcholine and catecholamines upon electrical stimulation was achieved in glands in which the endogenous acetylcholine stores in the splanchnic nerve terminals had been prelabelled by perfusion with [3H]choline. While apamin enhanced more than 2-fold the postsynaptic release of catecholamines, the presynaptic release of acetylcholine remained unaffected. 7. The results are compatible with the hypothesis that, under physiological conditions, Ca(2+)-activated SK channels present in chromaffin cells control the firing patterns of action potentials induced by the acetylcholine released from splanchnic nerves during stress.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 7473209 TI - A-current modifies the spike of C-type neurones in the rabbit nodose ganglion. AB - 1. In the rabbit nodose ganglion, C-type fibre neurones (C neurones) can be divided into two subtypes according to their after-hyperpolarizing potential (AHP) i.e. those with a fast AHP only and those with a fast AHP and a subsequent slow AHP produced by a slow calcium-dependent potassium current. In addition we have shown that some C neurones can be divided into two groups according to the effect of membrane hyperpolarization on their spikes i.e. type 1 in which duration and amplitude do not change and type 2 in which duration and amplitude decrease significantly. 2. In the present report we studied the effect of A current (IA) on spike duration, amplitude and slow AHP using intracellular recording techniques. 3. To detect the presence of IA, we first applied a series of increasing rectangular hyperpolarizing pulses to remove IA inactivation and then a short depolarizing pulse to trigger a spike. In type 1 C neurones the lag time of the spike in relation to hyperpolarization remains constant whereas in type 2 C neurones the spike only appears after IA inactivation and lag time in relation to hyperpolarization is lengthened. Thus, type 2 C neurones have an IA while type 1 C neurones do not. The fact that addition of cadmium did not change the lag time in type 2 C neurones shows that the IA is not calcium dependent. 4. Nodose neurones can be orthodromically activated by stimulation of the vagal peripheral process. In this way, after a hyperpolarizing pulse, IA can be fully activated by the orthodromic spike itself. Under these conditions it is possible to analyse the effects of IA on the spike. This was done by increasing either the hyperpolarizing potential, pulse duration, or the delay of the spike after the end of the pulse. We observed that maximum IA inactivation removal was always associated with the lowest duration and amplitude of the spike. 5. When IA inhibitors, 4-aminopyridine (4-AP) or catechol, were applied to type 2 C neurones, the delay of the spike after the hyperpolarization-depolarization test was no longer observed. In addition 4-AP abolished the shortening of the duration of the spike induced by steady hyperpolarization. 6. In type 2 C neurones with slow AHP, the IA-related decrease in spike duration was associated with a disappearance of the slow AHP. This indicates that IA decreases the calcium influx during the spike.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 7473210 TI - Different types of ganglion cell in the cardiac plexus of guinea-pigs. AB - 1. Intracellular recordings were made from the parasympathetic ganglion cells that lie in the epicardium of the left atrium of guinea-pig heart near the interatrial septum. 2. Three distinct types of neurone were identified on the basis of their electrophysiological properties. In one group of neurones, S cells, somatic action potentials were followed by brief after-hyperpolarizations. In the other two sets of neurones, somatic action potentials were followed by prolonged after-hyperpolarizations. The neurones with prominent after hyperpolarization were further subdivided: one group of neurones, P cells, showed inward rectification at membrane potentials near the resting membrane potential whilst neurones in the other group, SAH cells, did so only at more negative potentials. 3. In the group of neurones that displayed inward rectification at potentials near rest, rectification resulted from the activation of an inward current, which resembled the hyperpolarization-activated inward current present in cardiac muscle pacemaker cells. 4. The three different types of neurone received different patterns of synaptic input. Each SAH cell received a synaptic excitatory connection from the vagus which in most cells released sufficient transmitter to initiate an action potential in that cell; several SAH cells also received a separate connection, which could be activated by local stimulation. Although most S cells failed to receive a synaptic input from the vagus, all of those tested received an excitatory synaptic input which could be activated by local stimulation. Virtually all P cells failed to receive a synaptic input from the vagus; in addition, local stimulation failed to initiate synaptic potentials in P cells. 5. When the structure of cardiac ganglion cells was determined, by loading the cells with either biocytin or neurobiotin, it was found that most cells lacked extensive dendritic processes. S cells were invariably monopolar, most P cells were dipolar or pseudodipolar, whereas many SAH cells were multipolar. 6. In many neurones an on-going discharge of action potentials was detected in the absence of obvious stimulation. In S and SAH cells, the action potentials resulted from an on-going discharge of excitatory synaptic potentials. However, when a spontaneous discharge of action potentials was detected in P cells a discharge of excitatory synaptic potentials was not detected. 7. The results are discussed in relation to the idea that the three different types of cell may have different functions and that some of the cells may be organized in such a way as to permit the local handling of neuronal information within the heart. PMID- 7473211 TI - Neuropeptide gene expression and capsaicin-sensitive primary afferents: maintenance and spread of adjuvant arthritis in the rat. AB - 1. Many experimental and clinical arthritides are characterized by their bilateral nature. There is strong evidence to suggest that this bilateral spread may be mediated by a neuronal mechanism. We have previously shown early and sustained induction of mRNAs encoding preprotachykinin (PPT) and calcitonin gene related peptide (CGRP) in dorsal root ganglion (DRG) neurons innervating an inflamed, arthritic joint. We have now investigated the involvement of capsaicin sensitive primary afferents and the expression of neuropeptide mRNAs in the maintenance and bilateral spread of mild adjuvant-induced arthritis in the rat. 2. Capsaicin was applied perineurally to either the left (Cap-L) or right (Cap-R) sciatic nerve of halothane-anaesthetized male Han Wistar rats. Two weeks after capsaicin lesioning, arthritis was induced by injection of Freund's complete adjuvant (FCA) around the left ankle at a dose that caused inflammation of the left ankle joint, and a delayed (14 days) contralateral (right) ankle arthritis. Arthritis was monitored for 15 days after injection, when animals were killed and the lumbar DRG dissected. PPT, CGRP, somatostatin (SS), and vasoactive intestinal polypeptide (VIP) mRNA expression was determined in L5 DRG using in situ hybridization. 3. Spread of inflammation/arthritis to the right limb was associated with bilateral rises in PPT and CGRP mRNA expression in L5 DRG. SS mRNA expression in right DRG was unaffected by spread of inflammation. FCA-L+Cap L reduced left joint swelling and prevented spread of arthritis to the right joint when assessed by joint swelling. This inhibition of spread of arthritis was associated with significant reductions in all left L5 DRG neuropeptide mRNAs compared with controls, and the rise in right L5 DRG PPT mRNA expression seen in FCA-L-alone animals was blocked. FCA-L+Cap-R also reduced left joint swelling and prevented the spread of inflammation to the right ankle. This lesion prevented the rise in PPT and CGRP mRNA expression seen in right DRG with FCA-L alone. 4. These findings suggest a role for capsaicin-sensitive primary afferents and the primary afferent neuropeptides encoded by PPT and CGRP mRNA in the maintenance and spread of arthritis. PMID- 7473212 TI - Chemical mediators of spinal inhibition of rat sympathetic neurones on stimulation in the nucleus tractus solitarii. AB - 1. This study was undertaken to gain more direct evidence of the pathways and neurochemical mediators of a spinally mediated baroreceptor inhibition of sympathetic preganglionic neurones (SPNs). 2. For this purpose, single-pulse electrical stimulation within identified vasodepressor regions of the nucleus tractus solitarii (NTS) was used together with extracellular recordings of single antidromically identified SPNs in the T2 segment of the spinal cord of anaesthetized rats. 3. The actions of agonists and antagonists of inhibitory amino acids on the NTS-induced inhibitions were determined, when they were iontophoretically applied in the vicinity of SPNs via a multibarrel micropipette assembly. 4. Extracellular recordings were made from sixty-nine SPNs. In forty four SPNs, NTS stimulation elicited a period of inhibition of activity in both spontaneous and 'D,L-homocysteic acid-driven' SPNs with a latency to onset of 60 +/- 6 ms and a magnitude of 80 +/- 3%. 5. In six out of eight neurones, the NTS induced inhibition was reduced by 74 +/- 16% during the application of the glycine antagonist strychnine (0-10 nA, 5-10 min) with doses that selectively blocked the inhibitory effect of iontophoretically applied glycine. 6. In nine out of nine neurones, the NTS-induced inhibition was reduced by 38 +/- 6% during the application of the GABAA antagonist bicuculline (5-15 nA, 4-14 min) with doses that selectively blocked the inhibitory effect of iontophoretically applied GABA. 7. In two SPNs, the actions of strychnine and bicuculline were shown to be additive in blocking the NTS inhibition. 8. The selective GABAB antagonists, CGP 35348 (20-50 nA, 6-25 min) and CGP 55845A (10 nA, 11 min) did not antagonize the NTS-induced inhibition. 9. It is suggested that GABA and glycine interneurones are activated by a baroreceptor bulbospinal pathway to inhibit sympathetic preganglionic neurones in the spinal cord. PMID- 7473213 TI - Nitric oxide as a retrograde messenger in the nucleus tractus solitarii of rats during hypoxia. AB - 1. We examined the role of nitric oxide (NO) in respiratory regulation in the nucleus tractus solitarii (NTS), where L-glutamate release associated with peripheral chemoreceptor activation modulates the hypoxic ventilatory response. 2. Experiments were performed in unanaesthetized freely moving rats. First, the effects on the hypoxic ventilatory response of sodium nitroprusside (SNP, a NO donor) or NG-monomethyl-L-arginine (L-NMMA, a NO synthase inhibitor), microinjected into the NTS, were investigated. Second, using in vivo microdialysis, changes in extracellular L-glutamate during hypoxia were examined in the presence of L-NMMA. Third, the effect of L-NMMA on ventilatory augmentation by exogenous L-glutamate was examined. Furthermore, we measured extracellular L-citrulline concentration changes during hypoxia in the NTS to assess NO formation indirectly and also examined the effect of MK-801 (an NMDA receptor antagonist) on L-citrulline levels during hypoxia. 3. SNP increased ventilation during both normoxia and hypoxia. L-NMMA did not alter ventilation or L-glutamate levels during normoxia but significantly attenuated the hypoxic ventilatory response and the increase in L-glutamate during hypoxia. The inhibition by L-NMMA was blocked by L-arginine. The ventilatory augmentation by exogenous L-glutamate was attenuated by L-NMMA. L-Citrulline increased during hypoxia, and this increase was inhibited by MK-801. 4. We provide the first in vivo evidence that, in the NTS, NO works as a retrograde messenger in an L glutamate-releasing positive feedback system contributing to the augmentation of ventilation during hypoxia. PMID- 7473215 TI - Acceleration of puberty onset in female mice by male urinary proteins. AB - 1. Puberty onset in female mice is accelerated by exposure to conspecific adult male urine, which acts through the vomeronasal organ and the accessory olfactory system. A distinctive component of adult male mouse urine is the major urinary protein complex (MUP), which is a lipocalin; it has a hydrophobic pocket that binds small endogenous volatile molecules. The MUP gene family also codes for a hexapeptide, which has four residues in common with the N-terminal region of MUP. 2. MUP, the volatiles bound to MUP and the MUP-related hexapeptide have been tested for the induction of puberty acceleration by measuring the increase in uterus weight related to the first pro-oestrus phase. MUP, together with its bound volatiles, induces puberty acceleration. Its activity is retained even when the volatiles have been removed either by organic extraction or competition displacement with a high-affinity ligand. 3. MUP-related hexapeptide also induces puberty acceleration in female mice. In contrast, the odorants bound to MUP do not exert this effect. 4. It is proposed that the vomeronasal organ contains receptors that recognize the short N-terminal consensus sequence, N-Glu-Glu-Ala-X Ser (where X is a polar residue), common to both MUP and the hexapeptide. PMID- 7473214 TI - Visceral vasodilatation and somatic vasoconstriction evoked by acid challenge of the rat gastric mucosa: diversity of mechanisms. AB - 1. Acid back-diffusion through a disrupted gastric mucosal barrier increases blood flow to the stomach without any change in systemic blood pressure. This study was undertaken to examine the gastric acid-evoked changes in blood flow in a number of visceral and somatic arterial beds and to elucidate the mechanisms which lead to the regionally diverse haemodynamic responses. 2. The gastric mucosa of urethane-anaesthetized rats was challenged with acid by perfusing the stomach with ethanol (15%, to disrupt the gastric mucosal barrier) in 0.15 M HCl. Blood flow was estimated by laser Doppler flowmetry, the hydrogen clearance method or the ultrasonic transit time shift technique. 3. Gastric acid challenge increased blood flow in the gastric mucosa and left gastric artery while blood flow in the femoral artery and skin declined. 4. Afferent nerve stimulation by intragastric administration of capsaicin enhanced blood flow in the left gastric artery but did not diminish blood flow in the femoral artery when compared with the vehicle. 5. The gastric acid-evoked dilatation of the left gastric artery was depressed by acute extrinsic denervation of the stomach, capsaicin-induced ablation of afferent neurones or hexamethonium-induced blockade of autonomic ganglionic transmission. 6. The gastric acid-induced constriction of the femoral artery was attenuated by acute extrinsic denervation of the stomach but left unaltered by capsaicin, hexamethonium, guanethidine, indomethacin, telmisartan (an angiotensin II antagonist), [d(CH2)5(1), Tyr(Me)2, Arg8]-vasopressin (a vasopressin antagonist), bosentan (an endothelin antagonist) and acute ligation of the blood vessels to the adrenal glands. 7. These data show that acid challenge of the gastric mucosa elicits visceral vasodilatation and somatic vasoconstriction via divergent mechanisms. The gastric hyperaemia is brought about by extrinsic vasodilator nerves, whereas the reduction of somatic blood flow seems to be mediated by non-neural, probably humoral, vasoconstrictor messengers that remain to be identified. PMID- 7473216 TI - The effect of osmolality and carbohydrate content on the rate of gastric emptying of liquids in man. AB - 1. The effect of osmolality and carbohydrate content on the rate of gastric emptying was assessed by using the double sampling gastric aspiration technique to measure the rate of gastric emptying of isoenergetic and isosmotic solutions of glucose and glucose polymer. Six healthy male subjects were each studied on four separate occasions using a test drink volume of 600 ml. 2. The half-emptying time (t1/2, mean +/- S.E.M.) for a dilute (40 g l-1) solution of glucose (LG, 230 mosmol kg-1) was 17 +/- 1 min. This was greater than that (14 +/- 1 min) for a glucose polymer solution with the same energy content (LP, 42 mosmol kg-1). A concentrated (188 g l-1) glucose polymer solution (HP, 237 mosmol kg-1) emptied faster (t1/2 = 64 +/- 8 min) than the corresponding isoenergetic glucose solution (HG, 1300 mosmol kg-1, t1/2 = 130 +/- 18 min). 3. The dilute (40 g l-1) glucose solution emptied faster than the concentrated (188 g l-1) glucose polymer solution with the same osmolality (LG, 230 mosmol kg-1; HP, 237 mosmol kg-1). 4. The two dilute solutions (40 g l-1) delivered a similar amount of carbohydrate to the small intestine, whereas the concentrated (188 g l-1) glucose polymer solution delivered a greater amount of carbohydrate at 20, 40 and 50 min than the isoenergetic glucose solution. 5. These results indicate that both osmolality and carbohydrate content influence gastric emptying of liquids in man, but the carbohydrate content appears to have greater influence than osmolality.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7473217 TI - Modulation of the cGMP-gated ion channel in frog rods by calmodulin and an endogenous inhibitory factor. AB - 1. Outer segment patches excised in the light were used to investigate the effects of exogenous calmodulin and an endogenous inhibitory factor on the cGMP gated channel of frog rods. 2. Calmodulin shifted to the right the dose-response relation for activation of the channels by 8-Br-cGMP, but did not change the maximum current or the form of the relation. Reversal of this effect by removal of calmodulin was accelerated by brief exposure to saturating [8-Br-cGMP]. Inhibition by calmodulin required calcium and gave as much as a 5-fold decrease in current for an [8-Br-cGMP] functionally comparable to the presumed physiological [cGMP]. 3. Exposure to low [Ca2+]i (tens of nanomolar) appeared to irreversibly remove or inactivate an endogenous channel inhibitory factor from the patches, increasing the current at low [8-Br-cGMP]. Like calmodulin, this factor slowed the voltage-dependent channel-gating kinetics and did not change the maximum current. However, unlike calmodulin, the endogenous factor remained stably associated with the patches at high [Ca2+]i (1 microM), even with exposure to saturating [8-Br-cGMP]. 4. After the low-Ca2+ treatment increased the current, calmodulin reduced the current to about the same level as it had before the low Ca2+ treatment, giving a larger fractional suppression. Furthermore, patches with high initial sensitivity to 8-Br-cGMP had small low-Ca2+ effects and large calmodulin effects, while the reverse was true for patches with low initial agonist sensitivity. 5. Application of trypsin to the intracellular surface of the patch prevented the responses to calmodulin and to low [Ca2+]i, suggesting involvement of a cytoplasmic portion of the channel. However, trypsin also reduced the total agonist-induced patch current. 6. Our results are consistent with a model in which calmodulin and an endogenous calcium-binding protein compete for the same site, inhibiting channel opening or cGMP binding. The tight association of the endogenous factor with the channel even at relatively low [Ca2+]i suggests that in the transducing rod it may inhibit the channels most of the time in darkness and in dim light, preventing any potential inhibitory effects of calmodulin. The endogenous factor would be expected to leave the channel only in bright or prolonged light, when the [Ca2+]i is thought to be very low. PMID- 7473218 TI - Internal Ca2+ stores involved in anoxic responses of rat hippocampal neurons. AB - 1. During whole-cell recordings from CA1 neurons of rat brain slices with electrodes containing only KMeSO4 and Hepes, brief anoxia (2-3 min) consistently evoked a hyperpolarization (delta V approximately 14 mV) and reduction in input resistance (delta R approximately -20%). 2. As in previous intracellular recordings, Dantrolene sodium (10 microM) suppressed the anoxic delta V and delta R, confirming the release of internal Ca2+ is a major component of the anoxic response. 3. To identify the relevant intracellular Ca2+ store, other blockers of Ca2+ release were applied either externally (in the bath) or internally, by addition to the contents of the recording electrode. 4. The anoxic hyperpolarization was abolished or much reduced by heparin (10-20 micrograms ml 1, internal), thapsigargin (10 microM, external), Ruthenium Red (50 microM, internal) and external procaine (0.5-2 mM), but not by internal procaine (0.5-1 mM) or ryanodine (10 microM, external). 5. The anoxic fall in resistance was also abolished or reduced by heparin, thapsigargin and external procaine, but not by ryanodine, internal procaine or Ruthenium Red. 6. In addition, external procaine (0.5-2 mM) eliminated the early (transient) depolarization and reduced the post anoxic hyperpolarization by 60 +/- 22%. 7. None of these agents consistently changed the resting potential, but the input resistance was significantly increased by Dantrolene and external procaine. 8. In view of the marked effects of heparin and thapsigargin, but not ryanodine and internal procaine, we conclude that the anoxic response seen in such whole-cell recordings is initiated predominantly by Ca2+ release from an internal store that is InsP3 sensitive rather than Ca2+ sensitive. 9. Comparable but less pronounced effects of external procaine were seen during intracellular recordings with 3 M KCl-containing electrodes. The dose-dependent suppression of various features of the anoxic response by external procaine (EC50 approximately 0.2 mM) is presumed to be mediated by a superficial membrane trigger or modulating site. PMID- 7473219 TI - A highly calcium-selective cation current activated by intracellular calcium release in MDCK cells. AB - 1. The whole-cell patch clamp technique and fluorescence microscopy with the Ca2+ indicators fura-2 and fluo-3 were used to measure the whole-cell current and the free intracellular Ca2+ concentration ([Ca2+]i) in Madin-Darby canine kidney (MDCK) cells. 2. In a Ca(2+)-free bath solution, thapsigargin (TG) caused a transient increase of [Ca2+]i. Subsequent addition of Ca2+ caused a long lasting elevation of [Ca2+]i. 3. In a Ca(2+)-free bath solution, extracellular application of TG, ATP or ionomycin, or intracellular application of inositol 1,4,5-trisphosphate (IP3), caused a small but significant inward current (Iin) and a transient outward Ca(2+)-dependent K+ current (IK(Ca)), consistent with intracellular Ca2+ release. Subsequent addition of Ca2+ induced a prominent Iin with a current density of -4.2 +/- 0.7 pA pF-1. This Iin was unaffected by inositol 1,3,4,5-tetrakisphosphate (IP4). 4. Na+ replacement by mannitol, N methyl-D-glucamine+ (NMG+), aminomethylidin-trimethanol+ (Tris+) or choline+ reduced Iin by 54, 65, 52 and 56%, respectively. This indicates an apparent Ca2+ selectivity over Na+ of 26:1. Iin was, however, unaffected by replacing Cl- with gluconate- or by the K+ channel blocker charybdotoxin (CTX). 5. Iin was completely blocked by La3+ (IC50 = 0.77 microM). Consistently, La3+ completely reversed the TG-induced elevation of [Ca2+]i. SK&F 96365 (1-[3-(4-methoxyphenyl) propoxyl]-1-(4-methoxy-phenyl)-ethyl-1H-im idazole) HCl did not inhibit the TG induced Iin. It did, however, exhibit a biphasic effect on [Ca2+]i, consisting of an initial Ca2+ decay and a subsequent Ca2+ elevation. La3+ completely reversed the SK&F 96365-induced elevation of [Ca2+]i. 6. In the absence of Na+, Iin was dependent on the bath Ca2+ concentration (EC50 = 1.02 mM). Ca2+ replacement by Ba2+ or Mn2+ resulted in a reduction of Iin by 95 and 94%, respectively. 7. From these experiments we conclude that Ca2+ release from intracellular Ca2+ stores, induced by different independent methods, stimulates La(3+)-inhibitable Ca2+ entry in MDCK cells. Ca2+ entry is at least, in part, mediated by a cation current, which is highly, but not exclusively, selective for Ca2+ over Na+ and insensitive to SK&F 96365. PMID- 7473220 TI - Voltage-gated calcium currents in the magnocellular neurosecretory cells of the rat supraoptic nucleus. AB - 1. Whole-cell patch-clamp techniques were used to analyse voltage-dependent calcium currents in acutely isolated somata of magnocellular neurosecretory cells (MNCs) from the supraoptic nucleus of the hypothalamus of adult rats. Currents were characterized on the basis of their rates of inactivation and their sensitivity to a series of calcium channel blocking agents. 2. Curve fitting analysis of series of long lasting depolarizing voltage steps from a holding potential of -80 mV revealed three current components with different voltage dependences and rates of inactivation (n = 36). These include a low threshold ( 60 mV), rapidly inactivating (tau = 42 +/- 3 ms at -10 mV) component, a high threshold (-30 mV), slowly inactivating (tau = 1790 +/- 70 ms) component and a component with an intermediate threshold (-50 mV) and rate of inactivation (tau = 187 +/- 15 ms). There is also a non-inactivating portion of evoked calcium current with a threshold of -50 mV. 3. Based on its voltage dependence, rate of inactivation, greater sensitivity to the divalent cation nickel than to cadmium and insensitivity to omega-conotoxin GVIA (omega-CgTX), the low threshold current appears to be a T-type calcium current. The rate of inactivation, voltage dependence, and sensitivity to omega-CgTX of the slowly inactivating component suggests that it is an N-type current. The characteristics of the intermediate component do not correspond to any identified calcium current type. 4. Portions of the non-inactivating calcium current are sensitive to nifedipine (23 +/- 2% of the total non-inactivating current at -10 mV; n = 10), suggesting the presence of L-type currents, omega-agatoxin-IVA (omega-Aga-IVA; 20 +/- 6% of total; n = 11), suggesting the presence of P-type channels, and omega-CgTX (39 +/- 3% of total; n = 19), suggesting the presence of a non-inactivating N-type current. The non inactivating component at low potentials (> or = -50 mV) was selectively blocked by nifedipine, suggesting the presence of a novel, low threshold L-type current. 5. We conclude that MNC soma express T-, N-, L-, and P-type calcium currents, as well as a novel low threshold nifedipine-sensitive current and an unidentified inactivating component. This complement of currents is different from that seen in the terminals of these cells, suggesting a spatial and functional segregation of calcium current types in MNCs. PMID- 7473221 TI - Estimate of net calcium fluxes and sarcoplasmic reticulum calcium content during systole in rat ventricular myocytes. AB - 1. The experiments were performed on voltage-clamped cells in which intracellular calcium concentration ([Ca2+]i) was measured with the fluorescent indicator indo 1 (acetoxymethyl ester (AM) loading). When cells were stimulated with a short (100 ms) depolarizing pulse, following a rest, the magnitude of the first systolic calcium transient was greater than that in the steady state (rest potentiation) and decayed to its steady level over a few stimuli. If a longer pulse (800 ms) was used then the systolic calcium transient was either unaffected or increased in magnitude following a rest. During constant stimulation, if the length of the pulse is decreased, then the magnitude of the calcium transient decreased reversibly over several beats. 2. The calcium entry into the cell was measured from the integral of the inward calcium current and the efflux from the Na(+)-Ca2+ exchange current on repolarization. During the negative staircase the calcium current was approximately constant whilst the Na(+)-Ca2+ exchange current decayed in parallel with the systolic calcium transient. A net loss of calcium from the cell can be calculated from the extra Na(+)-Ca2+ exchange current following the initial pulses. 3. The application of caffeine produces a transient increase of both [Ca2+]i and an inward Na(+)-Ca2+ exchange current. The integral of this current can be used to estimate the caffeine-releasable calcium content of the sarcoplasmic reticulum (SR), which decreases following stimulation with short compared to long pulses. This difference in SR calcium content is quantitatively similar to that estimated from the sarcolemmal currents. 4. At a given membrane potential, the relationship between [Ca2+]i and current during the caffeine exposure can be used to estimate the Na(+)-Ca2+ exchange flux from the measured [Ca2+]i and thence the Na(+)-Ca2+ exchange flux during depolarization. 5. For a long depolarizing pulse the extrusion of calcium from the cell on Na(+) Ca2+ exchange is comparable to the entry on the calcium current. In contrast, for short pulses the extrusion of calcium on the Na(+)-Ca2+ exchange immediately after the pulse is greater than the entry during the pulse on the calcium current. 6. These results show that rest potentiation can be correlated with changes in the amount of calcium stored in the SR and this, in turn, can be accounted for by sarcolemmal fluxes. PMID- 7473222 TI - Activation mechanism of Ca(2+)-sensitive transient outward current in rabbit ventricular myocytes. AB - 1. The mechanism of activation of the Ca(2+)-sensitive and 4-aminopyridine (4-AP) insensitive transient outward current, I(to)(Ca), was examined in single rabbit ventricular myocytes using the whole-cell patch-clamp technique. 2. When the steady-state intracellular Ca2+ (Ca2+i) concentration ([Ca2+]i) was < 1 nM, I(to)(Ca) could not be activated by applying pulses at 0.1 Hz. When [Ca2+]i was increased to > or = 10 nM, I(to)(Ca) was activated by 0.1 Hz depolarizing pulses in all control experiments. 3. I(to)(Ca) was completely blocked by an anion transport blocker, DIDS, or by replacement of NaCl with sodium aspartate. Upon changing extracellular [Cl-], the reversal potential was shifted as predicted for a chloride-selective conductance. When intracellular K+ was replaced with Cs+, I(to)(Ca) was also observed. From these results it was concluded that I(to)(Ca) was carried by Cl-. 4. Anion selectivity of I(to)(Ca) was investigated by the replacement of C.- with various anions. The sequence of permeability was SCN- > I > Br- > Cl-. 5. The amplitude of I(to)(Ca) was enhanced in a [Ca2+]i-dependent manner between 10 nM and 1 microM Ca2+i, while steady-state inactivation curves and the voltage-dependent activation curves were unchanged. The half-inactivation and half-activation potentials were -35 mV and +37 mV, respectively, at all [Ca2+]i. 6. I(to)(Ca) was inhibited by blocking Ca2+ influx or Ca2+ release from sarcoplasmic reticulum, suggesting that a 'Ca(2+)-induced Ca(2+)-release' mechanism is essential for the activation of I(to)(Ca). 7. A steady-state Ca(2+) activated Cl- current with a linear I-V relationship was observed at 1 microM Ca2+, while the current activated by depolarization was strictly dependent on Ca2+ entry or Ca2+ release from the sarcoplasmic reticulum. These results suggest that the I(to)(Ca) channel is purely ligand (Ca2+) gated and its time course reflects the concentration of Ca2+i. PMID- 7473223 TI - Target-derived factors regulate the expression of Ca(2+)-activated K+ currents in developing chick sympathetic neurones. AB - 1. The functional expression of Ca(2+)-activated K+ currents (IK(Ca)) and voltage activated Ca2+ currents (ICa) was examined using whole-cell recordings from chick lumbar sympathetic neurones developing in situ and under various conditions in vitro. 2. Macroscopic IK(Ca) was expressed at low current density (< 0.01 mA cm 2) in neurones isolated at embryonic days 9-16 (E9-16). IK(Ca) was expressed at high densities (> 0.04 mA cm-2) at E17-19. By contrast, there was no significant difference in ICa density between sympathetic neurones isolated at E13 and E18. 3. When sympathetic neurones were isolated at E13 and maintained in vitro for 5 days, IK(Ca) was expressed at a significantly lower density (< 0.01 mA cm-2) than in neurones isolated acutely at E18 (> 0.04 mA cm-2). There was no difference in ICa density between neurones that developed in vitro and in situ. 4. When E13 sympathetic neurones were cultured for 5 days in the presence of a confluent layer of ventricular myocytes, they expressed IK(Ca) at a high density (> 0.04 mA cm-2), similar to that of E18 neurones that developed entirely in situ. Cardiac cell-conditioned medium produced similar effects. However, co-culture of sympathetic neurones with spinal cord explants did not allow for normal IK(Ca) expression in vitro. 5. Culturing sympathetic neurones in the presence of 5 ng ml 1 nerve growth factor (NGF) caused a significant increase in IK(Ca) density but this effect was only seen in 50% of cells examined. 6. The largest developmental changes in macroscopic IK(Ca) occur several days after other K+ currents and ICa are expressed at maximal density. The normal developmental expression of IK(Ca) is dependent upon extrinsic factors, including target-derived differentiation factors. PMID- 7473224 TI - Characteristics of Na(+)-Ca2+ exchange in frog skeletal muscle. AB - 1. Fluxes studies were carried out to investigate the Na(+)-dependent outward movement of Ca2+ in intact frog sartorius muscle from Leptodactylus ocellatus, a preparation for which published data on the subject are sparse. 2. Under normal resting conditions the Na(+)-Ca2+ exchange was not readily detectable. 3. When muscles were exposed to 4 mM caffeine, the rate of fractional loss of Ca2+ (kCa,o) increased by about 50%. Most of this increase exhibits characteristics typical of the Na(+)-Ca2+ antiport working in the forward mode found in other cells. 4. The increase in kCa,o promoted by caffeine was decreased by: (a) 72% in the absence of external Na+ (Nao+); (b) 73% in Na(+)-loaded muscles ([Na+]i = 98 mM); (c) 70% when fibres were depolarized to -27 mV ([K+]o = 50 mM); and (d) 80% in the presence of 5 mM amiloride. 5. Ni2+ (5 mM), an inhibitor of the Na(+)-Ca2+ exchanger current, unexpectedly increased the caffeine-promoted rise in kCa,o. This effect of Ni2+ was associated with a concomitant caffeine-stimulated Ni2+ influx. In the absence of caffeine Ni2+ did not affect kCa,o. 6. It was concluded that: (a) under resting conditions the sarcolemmal Ca2+ pump suffices to handle the cytosolic calcium concentration ([Ca2+]i); (b) Na(+)-Ca2+ activity becomes apparent when [Ca2+]i is substantially increased by caffeine-induced Ca2+ release from the sarcoplasmic reticulum; and (c) the blocking effect of Ni2+ on the current generated by a Na(+)-Ca2+ exchange with a coupling ratio > 2 might actually represent a shift of the antiport mode toward an electroneutral 1 Ni(2+) 1Ca2+ exchange. PMID- 7473225 TI - Modification by protons of frog skeletal muscle KATP channels: effects on ion conduction and nucleotide inhibition. AB - 1. The molecular mechanisms underlying pH regulation of skeletal muscle ATP sensitive K+ (KATP) channels were studied using the patch clamp technique in the inside-out configuration. Two effects of intracellular protons were studied in detail: the decrease in magnitude of single-channel currents and the increase in open probability (Po) of nucleotide-inhibited channels. 2. The pH dependence of inward unit currents under different ionic conditions was in poor agreement with either a direct block of the pore by protons or an indirect proton-induced conformational change, but was compatible with the protonation of surface charges located near the cytoplasmic entrance of the pore. This latter electrostatic mechanism was modelled using Gouy-Chapman-Stern theory, which predicted the data accurately with a surface charge density of about 0.1 negative elementary charges per square nanometre and a pK (pH value for 50% effect) value for protonation of these charges of 6.25. The same mechanism, i.e. neutralization of negative surface charges by cation binding, could also account for the previously reported reduction of inward unit currents by Mg2+. 3. Intracellular alkalization did not affect Po of the KATP channels. Acidification increased Po. In the presence of 0.1 mM ATP (no Mg2+), the channel activation vs. pH relationship could be fitted with a sigmoid curve with a Hill coefficient slightly above 2 and a pK value of 6. This latter value was dependent on the ATP concentration, decreasing from 6.3 in 30 microM ATP to 5.3 in 1 microM ATP. 4. Conversely, the channel inhibition vs. ATP concentration curve was shifted to the right when the pH was lowered. At pH 7.1, the ATP concentration causing half-maximal inhibition was about 10 microM. At pH 5.4, it was about 400 microM. The Hill coefficient values remained slightly below 2. Similar effects were observed when ADP was used as the inhibitory nucleotide. 5. These results confirm that a reciprocal competitive link exists between proton and nucleotide binding sites. Quantitatively, they are in full agreement with a steady-state model of a KATP channel possessing four identical protonation sites (microscopic pK, 6) allosterically connected to the channel open state and two identical nucleotide sites (microscopic ATP dissociation constant, approximately 30 microM) connected to the closed state. PMID- 7473226 TI - beta-adrenergic and cholinergic modulation of the inwardly rectifying K+ current in guinea-pig ventricular myocytes. AB - 1. Whole-cell patch-clamp technique was used to study the beta-adrenergic and cholinergic regulation of the inwardly rectifying K+ conductance (gK1) in isolated guinea-pig ventricular myocytes. 2. In Cl(-)-free solutions or in the presence of 9-anthracenecarboxylic acid or Co2+, bath-applied isoprenaline (Iso) partially inhibited the steady-state whole-cell conductance (gss) calculated from the steady-state current (Iss)-voltage (Iss-V) curve at membrane voltages (Vm) negative to the equilibrium potential for potassium (EK). Iss was also inhibited at Vm positive to EK when the extracellular [K+] was 20 mM. The Iso-sensitive component of gss exhibited the characteristics of the inwardly rectifying K+ conductance (gK1). 3. The Iso-induced inhibition of gK1 was reversible, concentration dependent, blocked by propranolol, mimicked by both forskolin and dibutyryl cAMP, and prevented by including a cAMP-dependent protein kinase (PKA) inhibitor in the pipette solution. These findings suggest that PKA mediates the Iso-induced inhibition of gK1. 4. The apparent dissociation constant (KD) for the concentration dependence of Iso-induced inhibition was 0.035 microM and the Hill coefficient was approximately 1.0. A maximal Iso concentration (1 microM) inhibited gK1 by 40 +/- 4.1% (mean +/- S.E.M.; n = 13). 5. Bath application of acetylcholine (ACh, 0.1 microM or more) antagonized the Iso-induced (1 microM) inhibition of gK1; [ACh] > 1.0 microM antagonized 88 +/- 2.1% (n = 10) of the inhibition. ACh increased the KD for Iso to inhibit Iso-sensitive gK1 and also reduced the maximal Iso-induced inhibition. 6. ACh-induced antagonism could be abolished by pre-incubating myocytes with pertussis toxin (PTX), suggesting that a muscarinic receptor-coupled, PTX-sensitive G protein, Gi, is involved. 7. ACh (10 microM) also antagonized approximately 70% of the dibutyryl cyclic AMP (1 mM) induced inhibition of gK1 (n = 3), suggesting that the ACh-induced antagonism involves more than simply inhibiting the Iso-mediated activation of adenylyl cyclase via the activated Gi. 8. Intracellularly applied okadaic acid (OkA, 1 microM) did not alter gK1 (control = 134 +/- 5.1 nS vs. OkA = 136 +/- 6.1 nS), but the Iso-induced decrease in gK1 was less (P < 0.001) with OkA present (42.1 +/- 2.4 nS, n = 5) than when absent (54.0 +/- 2.2 nS, n = 10). However, ACh (10 microM) failed to antagonize Iso-induced inhibition with OkA present, suggesting involvement of a protein phosphatase. PMID- 7473227 TI - Beta-adrenergic and cholinergic modulation of inward rectifier K+ channel function and phosphorylation in guinea-pig ventricle. AB - 1. To clarify the nature of the inhibition of whole-cell inwardly rectifying K+ current (IK1) by isoprenaline (Iso) and its antagonism by acetylcholine (ACh), we studied the effects of Iso and ACh and their surrogates on single channel currents (iK1) carried by inwardly rectifying K+ channels in cell-attached and excised inside-out patches obtained from guinea-pig ventricular myocytes. 2. Bath application of Iso suppressed iK1 channel activity in cell-attached patches. This was inhibited by propranolol. Bath-applied forskolin or dibutyryl cAMP mimicked the effect of bath-applied Iso. 3. Exposure of the cytosolic face of inside-out patches to purified catalytic subunit of the cAMP-dependent protein kinase (PKA) also suppressed iK1 channel activity, mimicking the effect of bath-applied Iso on iK1 recorded from cell-attached patches. 4. When applied directly to cell attached patches via the patch pipette solution, ACh antagonized Iso-induced (1 microM applied via the bath) suppression of iK1 channels. In contrast, bath applied ACh (10 microM) partially antagonized the effect of low concentrations of Iso (e.g. < 50 nM) on iK1 channels in cell-attached patches but had no detectable effect when 1 microM or more Iso was used. 5. In myocytes pretreated with pertussis toxin (PTX), ACh failed to antagonize Iso-induced suppression of iK1 channels. When inside-out patches were used, bath-applied preactivated exogenous inhibitory G protein subunit, G1 alpha, antagonized the suppression of iK1 channels induced by bath-applied catalytic subunit of PKA (PKA-CS), suggesting that a PTX-sensitive G1 alpha mediates ACh-induced antagonism of Iso-induced suppression of iK1. 6. Neither GTP gamma S nor G1 alpha antagonized the suppression of iK1 produced by bath-applied PKA-CS in inside-out patches when okadaic acid was present in the bath. In addition, bath application of alkaline phosphatase also reactivated iK1 channels suppressed by PKA-CS. 7. Findings in guinea-pig ventricular myocytes suggest that iK1 can be suppressed by a PKA mediated phosphorylation of the iK1 channel occurring in response to Iso-induced beta-adrenergic receptor activation and that ACh can antagonize the suppression by mechanisms that involve both intracellular and membrane-delimited pathways. The membrane-delimited pathway appears to involve M2-cholinergic receptors, their associated G protein, G1, and a protein phosphatase, all located in the sarcolemma in close proximity to the involved iK1 channels. PMID- 7473228 TI - Slowing of shortening velocity of rat cardiac myocytes by adenosine receptor stimulation regardless of beta-adrenergic stimulation. AB - 1. Single ventricular myocytes were enzymatically isolated, incubated with the A1 purinergic and beta-adrenergic receptor-specific agonists N6-cyclopentyladenosine (CPA) and isoprenaline (Iso), and then rapidly skinned. Ca2+ sensitivity of isometric tension and unloaded shortening velocity (Vo) were measured, and protein kinase A (PKA)-specific phosphorylations of troponin I (TnI) and C protein were assessed by back-phosphorylation of cell suspensions with [gamma 32P]-ATP. 2. Isoprenaline treatment decreased the Ca2+ sensitivity of isometric tension relative to propranolol-treated controls, as did simultaneous stimulation with Iso and CPA (Iso + CPA). CPA alone had no effect on Ca2+ sensitivity. Vo was greater in Iso-treated cells than in paired controls, while Vo was significantly less than control in both Iso + CPA-treated and CPA-treated cells. 3. Phosphorylation of TnI and C-protein was increased by Iso treatment and also when Iso and CPA were simultaneously applied. CPA alone caused a significant decrease in the phosphorylation state of these two proteins. 4. From these results we conclude that A1-purinergic receptor stimulation does not inhibit beta-adrenergic receptor-mediated phosphorylation of myofilament proteins, nor does it alter the Ca2+ sensitivity of isometric tension at the level of the myofilaments. However, A1-receptor stimulation does decrease Vo at the level of the myofilaments by a mechanism that is independent of beta-adrenergically mediated phosphorylation of TnI and C-protein. PMID- 7473229 TI - Reduced effect of pH on skinned rabbit psoas muscle mechanics at high temperatures: implications for fatigue. AB - 1. Inhibition of actomyosin function by decreased pH has been proposed to account for much of the depression of muscle function during fatigue. The clearest support for this hypothesis has been from studies of skinned skeletal muscle fibre mechanics at low temperatures (< or = 15 degrees C). 2. We re-examined the effect of decreased pH (7.0-6.2) on skinned mammalian skeletal fibre mechanics at low (10 degrees C) and high (30 degrees C) temperatures, using recently developed protocols that allow reproducible mechanical data to be obtained at higher temperatures. 3. At 10 degrees C we duplicated previous observations of a significant inhibition of maximum shortening velocity (Vmax) and isometric tension (Po) by acidosis. In contrast, at the higher temperature, we found only a very minimal effect of acidosis on Vmax and a threefold reduction in the decrease in Po. 4. Thus at temperatures only slightly below physiological for mammalian skeletal muscle systems, pH plays a much less important role in the process of muscle fatigue at the cross-bridge level than has been suggested by data obtained at physiologically unrealistic temperatures. PMID- 7473230 TI - Effects of Ca2+ channel blockers on transmitter release and presynaptic currents at the frog neuromuscular junction. AB - 1. The effects of the calcium channel blockers, funnel-web spider toxin (FTX), omega-agatoxin IVA (omega-Aga IVA) and omega-conotoxin GVIA (omega-CgTX), were tested on transmitter release and presynaptic currents in frog motor nerve endings. 2. Evoked transmitter release was blocked by FTX (IC50 = 0.02 microliter ml-1) and omega-CgTX (1 microM) but was not affected by omega-Aga IVA (0.5 microM). When FTX (0.1 microliter ml-1) was assayed on spontaneous release either in normal Ringer solution or in low Ca(2+)-high Mg2+ solution, it was found not to affect miniature endplate potential (MEPP) amplitude but to increase MEPP frequency by approximately 2-fold in both conditions. 3. Presynaptic calcium currents (ICa), measured by the perineurial technique in the presence of 10 mM tetraethylammonium chloride (TEA) and 200 microM BaCl2 to block K+ currents, were blocked by omega-CgTX (5 microM), partially blocked by FTX (1 microliter ml-1) and not affected by omega-Aga IVA (0.5 microM). 4. The presynaptic calcium activated potassium current (IK(Ca)) measured by the perineurial technique in the presence of 0.5 microM 3,4-aminopyridine (DAP) to block voltage-dependent K+ currents, was strongly affected by charybdotoxin (ChTX) (300 nM) and completely abolished by BaCl2 (200 microM). This current was also blocked by omega-CgTX (5 microM) and by CdCl2 (200 microM) but was not affected by FTX (1 microliter ml 1). The blockade by omega-CgTX could not be reversed by elevating [Ca]o to 10 mM. 5. The results suggest that in frog synaptic terminals two omega-CgTX-sensitive populations might coexist. The transmitter release process seems to be mediated by calcium influx through a omega-CgTX- and FTX-sensitive population. PMID- 7473231 TI - Failure of P-glycoprotein (MDR1) expressed in Xenopus oocytes to produce swelling activated chloride channel activity. AB - 1. P-glycoprotein, the protein product of the multidrug resistance (MDR1) gene, has ATP-dependent transporter activity. It has been suggested that P-glycoprotein may also function as a volume-regulated chloride channel or chloride channel regulator. To assess the chloride channel function of P-glycoprotein, we examined swelling-activated chloride conductances in Xenopus oocytes injected with human MDR1 cRNA. 2. Functional expression of P-glycoprotein in Xenopus oocytes was confirmed using Western blot analysis and by assessing transport of the P glycoprotein substrate, calcein AM. 3. Endogenous, swelling-activated chloride conductances were virtually absent by the time P-glycoprotein expression was confirmed. Thus, this expression system afforded the advantage of assessing putative MDR1-associated chloride currents in the absence of background currents. 4. The currents activated by hypotonic shock (50%) in both MDR1-injected and control (water-injected) oocytes were not significantly different. The swelling response was due in part to the activation of a potassium-selective conductance which could be inhibited by barium. No chloride-selective currents were activated by hypotonic shock in the presence or absence of barium. Therefore, we conclude that P-glycoprotein expression does not produce a swelling-activated chloride conductance in the Xenopus oocyte expression system. PMID- 7473232 TI - Activation of a Cl- conductance by SCN- in single proximal tubule cells isolated from Rana temporaria. AB - 1. This study investigated the effect of the anion, SCN-, on the conductive properties of single proximal tubule cells isolated from frog kidney. Ionic currents were measured using the conventional whole-cell patch clamp technique. 2. Addition of SCN- to the bath solution alone had a biphasic effect; there was an initial rapid rise, followed by a slower secondary increase, in both outward and inward conductances. However, when SCN- was added to the bath in the presence of pipette SCN-, such that the concentration gradient for movement of SCN- into the cell was abolished, only the fast changes in conductance were observed. 3. Cells did not discriminate between cations and anions under nominally K(+)-free control conditions. However, in the presence of intracellular SCN- cells became anion selective. Taken together with the increased conductance, this suggests activation of a Cl- channel (gSCN). 4. Niflumic acid, a Cl- channel blocker, inhibited gSCN in a dose-dependent manner, with a half-maximal blocking concentration (Ki) of 28 +/- 2.8 microM and a Hill coefficient of 1.2 +/- 0.3 (n = 6). 5. The activation of gSCN by intracellular SCN- was dose dependent and showed positive co-operativity, with a Ki of 62.3 microM and Hill coefficient of 4.0. 6. Plasma and urine levels of SCN- range between 10 and 70 microM, thus the conductance described here may play a role in the regulation of Cl- handling by the kidney. PMID- 7473233 TI - Erosion of inhibition contributes to the progression of low magnesium bursts in rat hippocampal slices. AB - 1. Bathing slices of rat hippocampus in media containing no magnesium ions results in epileptic discharges that originate in hippocampal area CA3. These discharges increase in severity gradually over periods of hours. 2. The progression of epileptic activity was much slower than the equilibration of extracellular magnesium activity and the resulting increase in strength of monosynaptic NMDA receptor-mediated excitation. Its time course matched that of a progressive decrease in pharmacologically isolated, evoked GABAA receptor mediated inhibitory postsynaptic current (IPSC) in the CA3 pyramidal cells. Conductance decreased to 37 +/- 6% of control values after 4 h. Responses to exogenous GABA application decreased to 52 +/- 12%. 3. Maximal IPSC conductance in 0 mM extracellular Mg2+ ([Mg2+]o) also decreased gradually when epileptic activity was abolished by bath application of 20 microM 6-cyano-7 nitroquinoxaline-2,3-dione (CNQX) and 50 microM D-2-amino-5-phosphonovaleric acid (D-APV) throughout the 4 h incubation period. It reached 61 +/- 8% of control values, a significantly smaller decline than that seen after 4 h of epileptic activity. 4. The decrease in mean IPSC conductance only partially reversed when the recording electrode contained 100 mM Mg2+. Complete recovery of IPSC strength occurred when electrodes also contained either 50 mM MgATP or 20 mM BAPTA. Reintroduction of 1 mM [Mg2+]o rapidly abolished epileptic activity and caused a slow, partial increase in IPSC conductance. 5. In the presence of 1 mM [Mg2+]o, GABAA receptor-mediated inhibition had to decrease to 17 +/- 11% of control values, in the presence of 4-7 microM bicuculline, to reach threshold for epileptic activity. 6. These data demonstrate a postsynaptic decrease in GABAA receptor-mediated inhibition in the in vitro low magnesium model of epilepsy. We propose that the apparent leaching of intracellular Mg2+ ([Mg2+]i) from cells leads to loss of ATP and consequent partial dephosphorylation of the GABAA receptor and that this is exacerbated by epileptic activity. PMID- 7473234 TI - Rhythm generation in the segmented hindbrain of chick embryos. AB - 1. The embryonic hindbrain of chick is segmented until stage 24, when it starts to generate rhythmic activities in cranial nerves. In order to recognize a possible influence of segmentation on the organization of neuronal systems generating motor rhythms, the activity of trigeminal, facial, glossopharyngeal, vagal and hypoglossal nerves was studied during embryonic stages 24-36, by simultaneous recording of different cranial nerves in an isolated, superfused chick hindbrain preparation. 2. Highly correlated recurrent episodes of cyclical burst discharges occurred in all nerves studied (correlation coefficients, 0.8 +/ 0.1) throughout stages 24-36. 3. Such coactivation is unlikely to be due to monosynaptic connections between widely divergent premotor neurons and motoneurons, or between motoneurons themselves, because no short-term correlation was apparent in the millisecond range between activities of different motor nerves. 4. Complete transverse or midsagittal sectioning of the hindbrain disrupted coactivation of nerves located at distinct rostrocaudal levels or occupying an ipsi- or contralateral position, respectively, while sparing the ability of individual nerves to generate rhythmic activity. Each hindbrain segment thus contains bilaterally the motor nuclei together with their own rhythm generator. Coactivation of motor patterns appears to result from intersegmental and cross-median connections between these rhythm generators. 5. The results are in keeping with the hypothesis of a segmental organization of the primordial hindbrain rhythm generator and give further support to the early determination of both the anatomical and the functional fate of neurons in this region of the vertebrate central nervous system. PMID- 7473235 TI - Repetitive firing properties of developing rat brainstem motoneurones. AB - 1. The repetitive firing properties of neonatal and adult rat hypoglossal motoneurones (HMs) were investigated in a brainstem slice preparation. Neonatal HMs could be classified into two main groups: (1) neurones with a decrementing or adapting firing pattern (type D); exhibiting an early and a late phase; and (2) neurones with an incrementing or accelerating firing pattern (type I). 2. The pattern of repetitive firing changed markedly during development. While most HMs recorded from young rats (< postnatal day (P) 4) were type D, the majority of HMs recorded during the second postnatal week were type I. In adults (> P21), nearly all HMs had a decrementing firing pattern, characterized by a brief period of adaptation and high steady-state firing rates. 3. The calcium-dependent after hyperpolarization (AHP) was shortest in type I neonatal HMs, and decreased in amplitude during trains of action potentials (APs). In type D neurones, these same trains caused a slight enhancement of AHP amplitude. In adult HMs, with a decrementing firing pattern, trains of APs also caused summation of the AHP. 4. Type D neonatal HMs showed a progressive prolongation of the AP during repetitive firing. In contrast, type I neonatal HMs had almost no change in AP duration. In adult HMs the AP was short and experienced only a modest increase in duration during fast repetitive firing. 5. The function relating steady-state firing frequency to injected current (f-I curve) was linear. The mean steady-state f-I slope was significantly higher in neonates than in adults (approximately 30 vs. approximately 20 Hz nA-1), and was weakly correlated with input resistance. The f I slope was negatively correlated with AHP duration in neonatal HMs only. In addition, for a given AHP duration the slope was higher in neonatal HMs. 6. Two threshold behaviours were observed among neonatal HMs: (a) a progressive rhythmic firing threshold, and (b) a sudden transition from subthreshold to regular repetitive firing. Current threshold for repetitive firing was strongly correlated with cell input conductance. Type I neonatal HMs had higher minimal steady firing rates (fmin) than type D HMs. In neonates, fmin was strongly correlated with AHP duration. Adult HMs showed a weaker correlation between these two parameters, and fmin was higher than predicted by AHP duration. 7. In summary, HMs responded to depolarizing current pulses with different firing patterns during postnatal development.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 7473236 TI - Non-uniform conduction time in the olivocerebellar pathway in the anaesthetized cat. AB - 1. It has recently been demonstrated that conduction velocities of cerebellar climbing fibre afferents in the rat are tuned according to fibre length such that conduction time between their origin in the inferior olive and their target cortical Purkinje cells is constant. Here we have examined the situation in the cat, where individual climbing fibres are substantially longer. Complex spike responses of Purkinje cells located at various depths in the vermis (zones a and b) were evoked by electrical stimulation of olivocerebellar fibres close to their origin and were recorded either extra- or intracellularly. 2. The onset latencies of directly evoked complex spikes ranged from 2.6 to 6.9 ms. A consistent trend in each electrode penetration was that the complex spike latencies were longer for the superficially encountered cells (where olivocerebellar fibre length is greatest) and shorter for deeper cells (where olivocerebellar fibre length is shorter). 3. Linear regression analysis suggests that conduction time in olivocerebellar fibres in the cat is not fixed but varies linearly with conduction distance. Our findings would be consistent with a uniform conduction velocity in olivocerebellar fibres of about 6.6 m s-1. PMID- 7473238 TI - Evidence favouring different descending pathways to soleus motoneurones activated by magnetic brain stimulation in man. AB - 1. In resting subjects low-intensity magnetic stimulation of the brain evoked an inhibition of the soleus H reflex at short latency (conditioning-test interval, 2 to +1 ms) followed approximately 10 ms later by a period of facilitation. During voluntary dynamic or tonic plantar flexion the same stimulus evoked a facilitation with a shorter latency than the inhibition (conditioning-test interval, -5 to -1 ms). 2. At the onset of ramp-and-hold plantar flexion the short-latency facilitation was seen at lower intensities of stimulation than the long-latency facilitation in six of seven subjects. At rest and/or during tonic plantar flexion the opposite was observed in four of the subjects, whereas the two facilitations had approximately the same threshold in the remaining subjects. 3. The short-latency facilitation decreased approximately 100 ms after the onset of ramp-and-hold plantar flexion in all of eight subjects. The long-latency facilitation, in contrast, either had the same size throughout the ramp phase or even increased around the end of the ramp phase. 4. The short-latency facilitation of the reflex was significantly larger at the onset of a fast ramp and-hold plantar flexion (10 N m (150 ms)-1) than at the onset of a slow contraction (10 N m (600 ms)-1), whereas the opposite was the case for the long latency facilitation. 5. As the short- and long-latency facilitations had different thresholds and were differently regulated during voluntary movement, it is suggested that they are caused by activation of different descending pathways by the magnetic stimulus. PMID- 7473237 TI - Involvement of neurokinin receptors in the induction but not the maintenance of mechanical allodynia in rat flexor motoneurones. AB - 1. Intrathecal (i.t.) injections of the (tachykinin) NK1 receptor agonist, substance P methyl ester (SPME; 20 pmol), or the NK2 receptor agonist, neurokinin A (NKA; 20 pmol), substantially decreased the cutaneous mechanical threshold and markedly enhanced the touch-evoked response of posterior biceps femoris semitendinosus (PBF-ST) spinal flexor motoneurones in decerebrate-spinal rats. This cutaneous mechanical reflex allodynia was prevented by pretreatment with the NK1 antagonist RP 67580 (2.28 nmol, i.t.) and the NK2 antagonist MEN 10376 (0.7 nmol, i.t.), respectively. 2. Electrical stimulation of the sural nerve at C fibre strength or cutaneous application of the irritant, mustard oil, produced prolonged cutaneous mechanical allodynia in PBF-ST motoneurones (15 min and > 1 h, respectively). Pretreatment with RP 67580 but not MEN 10376 prevented this, but when RP 67580 was administered 25 min after the application of mustard oil, the established hypersensitivity of the flexor motor reflex was not reversed. The enantiomer of RP 67580, RP 68651 was without effect. 3. Injection of bradykinin (60 microM, 80 microliters) into the gastrocnemius muscle increased the cutaneous mechanical hypersensitivity of PBF-ST flexor motoneurones for 40-50 min. MEN 10376, but not RP 67580, prevented this, but only when administered prior to the bradykinin injection. 4. These results suggest that the induction, but not the maintenance, of cutaneous mechanical allodynia in flexor motoneurones is NK receptor dependent, with cutaneous C fibre conditioning inputs acting via NK1 and muscle C fibre conditioning inputs via NK2 receptor subtypes. PMID- 7473239 TI - Ingestion of branched-chain amino acids and tryptophan during sustained exercise in man: failure to affect performance. AB - 1. An increased uptake of tryptophan in the brain may increase serotoninergic activity and recently has been suggested to be a cause of fatigue during prolonged exercise. The present study, therefore, investigates whether ingestion of tryptophan or the competing branched-chain amino acids (BCAAs) affect performance. Ten endurance-trained male athletes were studied during cycle exercise at 70-75% maximal power output, while ingesting, ad random and double blind, drinks that contained 6% sucrose (control) or 6% sucrose supplemented with (1) tryptophan (3 g l-1), (2) a low dose of BCAA (6 g l-1) or (3) a high dose of BCAA (18 g l-1). 2. These treatments greatly increased the plasma concentration of the respective amino acids. Using the kinetic parameters of transport of human brain capillaries, BCAA supplements were estimated to reduce brain tryptophan uptake at exhaustion by 8-12%, while tryptophan ingestion caused a 7- to 20-fold increase. Exercise time to exhaustion was not different between treatments (122 +/- 3 min). 3. The data suggest that manipulation of tryptophan supply to the brain either has no additional effect upon serotoninergic activity during prolonged exhaustive exercise or that manipulation of serotoninergic activity functionally does not contribute to mechanisms of fatigue. PMID- 7473240 TI - Ryanodine receptor-mediated intracellular calcium release in rat cerebellar Purkinje neurones. AB - 1. Ryanodine receptor-mediated Ca2+ release was investigated in Purkinje neurones of rat cerebellar slices by using whole-cell patch-clamp recordings combined with fluorometric digital imaging of cytoplasmic Ca2+ concentration ([Ca2+]i). 2. Caffeine caused a transient increase in [Ca2+]i in the somata and dendrites of Purkinje neurones. Caffeine-induced Ca2+ transients were not associated with a membrane inward current and persisted in Ca(2+)-free external solutions, indicating that they are caused by Ca2+ released from intracellular stores. The amplitudes of the caffeine-mediated elevations in [Ca2+]i were strongly dependent on the baseline level of [Ca2+]i. 3. Intracellular application of Ruthenium Red through the patch pipette blocked caffeine-induced Ca2+ transients in Purkinje neurones. Ryanodine when applied either intra- or extracellularly caused a use dependent block of caffeine-induced Ca2+ release. 4. Depolarization-induced Ca2+ transients were strongly prolonged by caffeine. Several lines of evidence suggest that these prolongations reflect Ca(2+)-induced Ca2+ release. 5. Despite the presence of skeletal muscle type ryanodine receptors in Purkinje neurones, depolarizing pulses failed to induce any changes in [Ca2+]i when the influx of Ca2+ through voltage-gated channels was prevented by using Ca(2+)-free solution, or when applying blockers of voltage-gated Ca2+ channels. 6. Dendritic Ca2+ transients produced by stimulation of the excitatory climbing fibre synaptic input were also prolonged by caffeine, indicating that ryanodine receptor mediated release of Ca2+ may be involved in synaptic signalling in cerebellar Purkinje neurones. 7. Ryanodine receptor-mediated release of Ca2+ in cerebellar Purkinje neurones can be explained by a model in which release of Ca2+ is strongly facilitated by the co-operative action of Ca2+, caffeine and/or ryanodine. Our results suggest that Ca2+ release in these central neurones becomes prominent only during episodes of intensive electrical activity associated with increased Ca2+ entry. PMID- 7473241 TI - Different effects on gating of three myotonia-causing mutations in the inactivation gate of the human muscle sodium channel. AB - 1. Three mutations at the same site in the inactivation gate of the alpha-subunit of the human muscle Na+ channel, G1306E, G1306V and G1306A, cause three phenotypes of K(+)-aggravated myotonia: G1306E as the most severe and G1306A as the most benign form. 2. Recombinant wildtype (WT) and mutant (G1306E, G1306V and G1306A) human Na+ channels were expressed in human embryonic kidney cells (HEK293). G1306E and G1306V channels showed a distinct increase in the time constants of inactivation (tau h1 and tau h2) and in the ratios of steady-state to peak currents (Iss/Ipeak) (e.g. at 0 mV, G1306E vs. WT; tau h1, 1.29 +/- 0.10 vs. 0.52 +/- 0.01 ms; Iss/Ipeak, 2.90 +/- 0.40 vs. 0.93 +/- 0.19%). G1306A channels showed only an increase in tau h1 (0.74 +/- 0.07 ms). For G1306E and G1306V channels, the steady-state inactivation curves, as well as the voltage dependence of the rate of recovery from inactivation, were shifted by +15 mV. For G1306A the h infinity curve was shifted by only +5 mV. 3. G1306E and G1306V channels showed prolonged current rise times and later first openings suggesting slowing of activation. For G1306E channels only, the steady-state activation curve was shifted by -7 mV. For all mutants the deactivation time constants were increased. 4. We conclude that (i) the combination of alterations in inactivation and activation produces the slowing of the current decay, (ii) the slowed inactivation is most responsible for myotonia, and (iii) the shift of the steady state activation curve, seen only with G1306E channels, may explain the severity of this phenotype. 5. The results suggest that two of the mutations in the Na+ channel inactivation gate also alter channel activation and deactivation. PMID- 7473242 TI - Decreased calcium currents in motor nerve terminals of mice with Lambert-Eaton myasthenic syndrome. AB - 1. The effects of immunoglobulin G (IgG) from patients with Lambert-Eaton myasthenic syndrome on Ca2+ currents in mammalian motor nerve terminals are unknown. Therefore, we recorded these currents in phrenic nerves of mice injected with serum from either LEMS patients, myasthenia gravis patients, or healthy control individuals. 2. In control preparations, the endplate currents induced by repetitive stimulation at > or = 20 Hz were depressed as expected. However, in the LEMS animals quantal content decreased and either depression did not occur or synaptic facilitation occurred. 3. Ca2+ currents were smaller in LEMS animals. At 0.5 Hz stimulation frequency, normalized Ca2+ currents in LEMS animals were 57 +/ 14% of those in control. At higher frequencies, Ca2+ currents become smaller in control but not in LEMS animals. 4. Ca2+ currents in controls were unaffected by addition of nifedipine but were reduced by 37% upon addition of omega-conotoxin GVIA. In LEMS animals, however, the currents were depressed by 43% by nifedipine but were unaffected by omega-conotoxin GVIA. Thus, LEMS is associated with reduced Ca2+ currents and a shift to dihydropyridine sensitivity. PMID- 7473243 TI - Synaptic input from CA3 pyramidal cells to dentate basket cells in rat hippocampus. AB - 1. Excitatory inputs from CA3 pyramidal cells to dentate basket cells were examined using the whole-cell recording technique in neonatal (10-16 days) rat hippocampal slices to characterize this unexpected feedback pathway. 2. Minimal electrical stimulation of the CA3 pyramidal layer evoked in basket cells short latency (5.2 +/- 0.4 ms) glutamate receptor-mediated excitatory postsynaptic currents (EPSCs) with fast rise times (at -70 mV, 0.9 +/- 0.2 ms), fast decay time constants (3.6 +/- 0.6 ms), and small amplitudes (-14 +/- 3.4 pA). Minimal electrical stimulation evoked monosynaptic EPSCs in only 48 +/- 9.2% of the trials suggesting that the CA3 pyramidal cell to basket cell pathway was unreliable. 3. CA3 pyramidal cell layer stimulation did not antidromically or synaptically activate granule cells but did evoke polysynaptic IPSCs in granule cells, suggesting that the net effect of CA3 pyramidal cell firing on the dentate gyrus was granule cell inhibition. 4. Stimulation of the CA3 pyramidal cell layer evoked both monosynaptic and polysynaptic EPSCs in basket cells, which were eliminated by a knife lesion separating CA3 from the dentate gyrus. The latencies of the EPSCs evoked in 0.6 mM extracellular calcium were the same as the earliest latencies of EPSCs in 1.5 mM calcium, suggesting that those EPSCs were monosynaptic. The polysynaptic input was more prominent in the presence of 10 microM bicuculline, implying that inhibitory GABAergic circuits normally limit this feedback from CA3 to basket cells. 5. In recordings from 103 pairs of CA3 pyramidal cells and dentate basket cells from 11 slices, two polysynaptic connections were found that were active only when the presynaptic CA3 pyramidal neuron fired in bursts. No monosynaptic connections between CA3 pyramidal cells and basket cells were identified indicating that connections between the two cell types may be sparse. 6. Raising the external potassium concentration from 3.5 to 8.5 mM, which elicited burst firing in CA3 pyramidal cells, resulted in a barrage of EPSCs and action potentials in basket cells. In contrast, granule cells neither fired action potentials nor exhibited increased EPSC frequency in elevated potassium but instead received a higher frequency of bicuculline sensitive IPSCs, consistent with interneuron firing. The CA3 pyramidal cell to basket cell monosynaptic pathway exhibited paired-pulse facilitation as manifested by an increased probability of release, which supports the idea that basket cells were better activated by short trains of action potentials than by single inputs.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 7473244 TI - Effects of angiotensin II in fetal sheep and modification of its actions by indomethacin. AB - 1. Angiotensin II (AII) was infused I.V. into seven chronically catheterized fetal sheep (gestational age, 120-136 days). The effects of short-term infusions of 6 and 12 micrograms kg-1 h-1 for 1.5 h were compared with the effects of infusing 6 micrograms kg-1 h-1 for 3 or 5 days (long-term infusion). AII produced an immediate rise in fetal arterial blood pressure (P < 0.025). When infused for 3 or 5 days, 6 micrograms kg-1 h-1 AII caused a greater increase in arterial blood pressure (P < 0.05). 2. Infusions of 6 micrograms kg-1 h-1 AII for 1.5 h had no effect on fetal placental blood flow or on flow to the fetal membranes, but after AII infusion for 3 or 5 days both flows were reduced (P < 0.01 and P < 0.005, respectively). Fetal blood gas status and pH were maintained. The only change in fetal renal function observed with short-term infusions of AII was a rise in sodium excretion when 12 micrograms kg-1 h-1 AII was given (P < 0.05). Infusion of 6 micrograms kg-1 h-1 for 3 or 5 days also caused a rise in sodium excretion (P < 0.025) because total and proximal fractional sodium reabsorptions were depressed (P < 0.01). Infusions of AII had no effects on the volume of lung liquid produced or on its composition. 3. Administration of indomethacin to the ewe (10 mg kg-1) and to the fetus (12 mg kg-1), during the infusion of AII, caused a rise in maternal arterial pressure (P < 0.01) but no change in fetal arterial pressure. 4. After indomethacin, umbilicoplacental blood flow rose (P < 0.05), as did fetal arterial PO2 (P < 0.05). Fetal arterial PCO2, pH and bicarbonate levels fell (P < 0.01). Glomerular filtration rate (GFR) rose (P < 0.01); there was a natriuresis (P < 0.01), chloriuresis (P < 0.01) and a kaliuresis (P < 0.05) but urine flow rate did not change. Lung liquid flow fell (P < 0.01). 5. It is concluded that in the fetus, long-term infusions of AII at a constant dose rate cause a progressive rise in arterial pressure. In addition, effects of AII on placental blood flow and on renal function develop. Thus, short term infusions of AII cannot be used to predict the renal and cardiovascular effects of sustained high levels of this peptide in the fetus.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 7473245 TI - Concentrations of Na+ and Cl- in transplacental ultrafiltrate in sheep. AB - 1. Maternal blood was made transiently hypertonic by rapid I.V. infusion of a concentrated mannitol solution into pregnant ewes bearing lambs with an indwelling flow sensor and vascular catheters. 2. The transplacental flows of water and of Na+ and Cl- were calculated from the umbilical arteriovenous differences in the concentrations of 125I-labelled albumin and electrolytes, and the fetal placental blood flow. 3. The reflection coefficients of Na+ and Cl- were calculated by means of the Patlak equation and found to be 0.85 +/- 0.04 and 0.68 +/- 0.04 (means +/- S.E.M.). The filtration coefficient was 1.02 x 10(-7) +/ 0.12 x 10(-7) cm5 dyne-1 s-1. 4. The results fitted best to an equivalent pore radius in the placental barrier smaller than the currently accepted 0.44 nm but not less than 0.35 nm. PMID- 7473246 TI - Fluid uptake from the renal medulla into the ascending vasa recta in anaesthetized rats. AB - 1. We have investigated fluid movements between superficial ascending vasa recta (AVR) and the interstitium of exposed papillae of the renal medullae in 15-day old Sprague-Dawley rats anaesthetized with Hypnorm and Hypnovel. 2. Using a development of the red cell micro-occlusion technique, fluid filtration and reabsorption rates per unit area of vessel wall (Jv/A) were determined in 54 single perfused AVR at known microvascular pressures (Pc). The relation between Jv/A and Pc was non-linear suggesting hydraulic permeabilities (Lp) of 50-100 x 10(-7) cm s-1 cmH2O-1 when Pc was between 0-10 cmH2O and 150-200 x 10(-7) cm s-1 cmH2O-1 when Pc was 10-15 cmH2O. 3. Rates of fluid reabsorption into the AVR estimated by a densitometric technique in a further fourteen vessels were consistent with Lp values of 50-100 x 10(-7) cm s-1 cmH2O-1 when Pc was -2 to 0 cmH2O. 4. The effective oncotic pressures of perfusates containing bovine serum albumin (BSA) were consistent with minimum values for the reflection coefficients of the walls of the AVR to BSA of between 0.59 and 0.72. 5. The concentration of native serum albumin in the papillary interstitial fluid was 9.1 +/- 0.6 mg ml-1 (mean +/- S.E.M., n = 16, from 9 rats), which is approximately 25% of the plasma level. 6. After their microinjection into the medullary interstitium, Patent Blue V and Evans Blue-albumin cleared within 1 min. There was no evidence of preferential movement of either dye towards the base of the exposed renal medulla. 7. Because Lp of the AVR is high, mean pressures of only approximately 3 cmH2O are necessary to account for the total clearance of fluid from the medullary interstitium into the AVR. From published data and from our own observations, it appears that differences in hydrostatic and oncotic pressure across the walls of the AVR are more than sufficient to provide this driving force. The possibility of the clearance of protein from the interstitium into the AVR is discussed. PMID- 7473247 TI - The multi-ion nature of the cGMP-gated channel from vertebrate rods. AB - 1. Native cGMP-gated channels were studied in rod outer segments of the larval tiger salamander, Ambystoma tigrinum. The alpha-subunit of the cGMP-gated channel, here referred to as the wild type (WT), and mutant channels were heterologously expressed in Xenopus laevis oocytes. These channels were studied in excised membrane patches in the inside-out configuration and were activated by the addition of 100 or 500 microM cGMP. The current carried by monovalent cations was measured under voltage-clamp conditions. 2. In the presence of 110 mM Na+ in the extracellular medium and different amounts of Na+ in the intracellular medium, the I-V relations of the native channel could be described by a single site model with a profile of Gibbs free energy with two barriers and a well. A similar result was obtained in the presence of 110 mM Li+ in the extracellular medium and different amounts of Li+ in the intracellular medium. The well depth was 1.4RT (where R is the gas constant and T is the absolute temperature) for both Li+ and Na+. 3. The I-V relations of the native channel in the presence of 110 mM Na+ on one side of the membrane and 110 mM Li+ on the other side could not be described by the same single-site model with identical values of barriers and well obtained in the presence of Li+ or Na+ alone: the well for Li+ had to be at least 4RT. 4. In the presence of mixtures of 110 mM Li+ and Cs+ on the cytoplasmic side of the membrane, an anomalous mole fraction effect was observed both in the native and the WT channel. No anomalous behaviour was seen in the presence of Li(+)-Na+ and Li(+)-NH4+ mixtures. 5. The anomalous mole fraction effect with mixtures of Li+ and Cs+ was not observed in the channel where glutamate 363 was mutated to a glutamine (E363Q) or an asparagine (E363N). When glutamate 363 was mutated to an aspartate (E363D), the anomalous mole fraction effect with mixtures of Li+ and Cs+ was still observed, although significantly reduced. 6. When lysine 346, arginine 369, aspartate 370 and glutamate 372 were neutralized by mutation to glutamine, the ion permeation through the mutant channels and the WT channel had largely similar properties.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 7473248 TI - Modulation of Na(+)-H+ exchange by altered cell volume in perfused rat mandibular salivary gland. AB - 1. Intracellular pH (pHi) was measured by spectrofluorometry in perfused mandibular salivary glands isolated from the rat and loaded with the pH-sensitive fluoroprobe 2',7'-bis(2-carboxyethyl)-5(6)-carboxyfluorescein (BCECF). Cell volume changes were estimated from changes in intracellular water content measured by proton NMR spectroscopy. 2. Stimulation with 1 microM acetylcholine (ACh) led to a 15 +/- 2% decrease in cell volume. A transient decrease in pHi was followed by a sustained increase (0.17 +/- 0.03 pH units) that has previously been attributed to the upregulation of the Na(+)-H+ exchanger. 3. Increasing perfusate osmolarity by addition of 60 mM sucrose caused a 19 +/- 2% decrease in cell volume and a sustained increase in pHi (0.12 +/- 0.01 pH units) that was abolished by 1 mM amiloride. Acid loading experiments indicated that the increase in pHi was due to an alkaline shift in the pH dependence of the Na(+)-H+ exchanger. 4. A 20% reduction in perfusate osmolarity prevented the cell shrinkage normally associated with ACh stimulation and largely abolished the ACh induced increase in pHi. 5. Steady-state Na(+)-H+ exchanger activity, estimated from the initial rate of change in pHi following addition of amiloride, increased 9-fold during stimulation with ACh. When cell shrinkage was prevented by simultaneous exposure to the hypotonic solution, the activity of the exchanger still increased 7-fold in response to ACh. 6. We conclude that, although cell shrinkage leads to upregulation of the Na(+)-H+ exchanger, this factor alone is insufficient to account for the marked increase in exchanger activity that follows muscarinic stimulation. PMID- 7473249 TI - Ankle extensor group I afferents excite extensors throughout the hindlimb during fictive locomotion in the cat. AB - 1. The effects of stimulating hindlimb extensor nerves (100-200 ms trains, 100 Hz, < or = 2 times threshold) during the flexor and extensor phases of the locomotor step cycle were analysed in the decerebrate, paralysed cat during fictive locomotion evoked by stimulation of the mesencephalic locomotor region. 2. Stimulation during extension of either the medial gastrocnemius (MG), lateral gastrocnemius-soleus (LGS) or plantaris (Pl) nerves was equally effective in increasing the duration and amplitude of electroneurogram (ENG) activity recorded in ipsilateral ankle, knee and hip extensor nerves. Enhancement of extensor ENG activity could be evoked with near threshold stimulation intensity and appeared within 10-40 ms of the onset of ankle extensor nerve stimulation. Stimulation of anterior biceps during extension occasionally evoked a modest increase in the duration of activity of hip, knee and ankle extensors. Stimulation of quadriceps during extension enhanced the activity of proximal extensors and soleus, but inhibited other ankle extensors. 3. Selective activation of ankle extensor Ia spindle afferents by muscle stretch also enhanced ipsilateral extension. It is argued that both muscle spindle and tendon organ afferents can contribute to the increase in extensor nerve activity evoked by group I stimulation intensity during fictive locomotion. 4. During flexion, stimulation of either the MG, Pl or LGS nerves at group I strength terminated on-going activity in ipsilateral flexors and initiated a burst of activity in ipsilateral hip, knee and ankle extensors, i.e. reset the step cycle to extension. 5. Low strength stimulation of the mixed muscle and cutaneous nerve innervating the plantar aspect of the foot produced extension enhancement and resetting similar to that evoked by group I muscle afferent stimulation. Stimulation of the cutaneous nerve supplying the dorsal aspect of the foot during extension enhanced extensor activity, and during flexion, enhanced the activity of flexors. 6. The effects reported here during fictive locomotion may also occur during overground locomotion with natural activation of group I muscle spindle and tendon organ afferents. Extensor spindle and tendon organ afferents may thus serve as an excitatory reflex system helping to shape the amplitude, duration and timing of ipsilateral extensor activity. Increased or unexpected activation of group I ankle extensor afferents or plantar foot afferents during locomotion could also compensate for increased loading of the limb. PMID- 7473250 TI - Effects of stimulation of hindlimb flexor group II afferents during fictive locomotion in the cat. AB - 1. This study examines the effects of electrical stimulation of hindlimb flexor nerves on the fictive locomotion pattern. Locomotion was initiated by stimulation of the mesencephalic locomotor region in the decerebrate paralysed cat and monitored by recording the electroneurogram from selected hindlimb flexor and extensor muscle nerves. Flexor nerves were stimulated using short trains (20-50 stimuli at 100 Hz) during either the flexor or the extensor phase of the fictive locomotor cycle. 2. Stimulation of tibialis anterior (TA), posterior biceps and semitendinosus (PBSt) or sartorius (Sart) nerves at 5 times threshold (T) during the flexor phase of the fictive locomotor cycle terminated on-going activity in flexor nerves and initiated activity in extensors. Thus, flexor nerve stimulation during flexion shortened the locomotor cycle by resetting to extension. The failure of lower intensity (2T) stimulation of PBSt or Sart nerves to reset the step cycle to extension suggests that group II afferents are responsible for these actions. Resetting evoked by 2T stimulation of the TA nerve may be due to a high proportion of group II afferents with low electrical threshold. 3. During extension, stimulation of TA and PBSt nerves at 5T did not perturb the locomotor rhythm whereas Sart stimulation prolonged the locomotor cycle. 4. Stimulation of cutaneous or knee joint afferents failed to produce effects similar to those evoked by stimulation of flexor muscle nerves at group II strength. These findings are at odds with those obtained elsewhere in the acute spinal, DOPA fictive locomotion preparation. The possibility that group II resetting during fictive locomotion is not mediated by flexion reflex pathways but by previously unknown pathways released in the present preparation is discussed. 5. Since many of the flexor afferents recruited by 5T electrical stimulation are the length sensitive group II fibres, spindle secondaries may act to regulate the duration and onset of flexor and extensor activity during real locomotion. The resetting from flexion to extension also suggests that unexpected or enhanced activity of flexor secondaries during swing would promote a switch of the step cycle to stance. PMID- 7473251 TI - Reciprocal inhibition between wrist flexors and extensors in man: a new set of interneurones? AB - 1. Interneurones mediating reciprocal inhibition between wrist flexors and extensors in man are characterized using both Renshaw cells and transarticular group I afferent activation. 2. Renshaw cells were activated by reflex discharges evoked by a tendon tap. The tendon tap was applied to the tendon of the muscles from which the Ia fibres responsible for the reciprocal inhibition originated. Contrary to what was observed both in the cat hindlimb and in human elbow muscles, this Renshaw cell activation never resulted in a long depression of the reciprocal inhibition between wrist flexors and extensors. 3. Convergence from group I elbow muscle afferents and antagonistic group I afferents onto interneurones mediating reciprocal inhibition between wrist muscles was revealed in post-stimulus time histogram (PSTH) experiments using the technique of spatial facilitation. 4. The characteristics of the interneurones mediating reciprocal inhibition between wrist flexors and extensors could therefore be summarized as follows: (a) they are fed by antagonistic group I afferents and group I afferents originating from both flexor and extensor elbow muscles; (b) they are not inhibited by Renshaw cells; (c) they are not excited by low threshold cutaneous afferents; and (d) they are probably interposed in a disynaptic pathway. 5. It is therefore concluded that interneurones mediating reciprocal inhibition between wrist flexors and extensors in man differ both from Ia interneurones and from interneurones interposed in the Ib reflex pathways and these characteristics are related to the complex circumduction movements developed in the wrist. PMID- 7473252 TI - Cutaneomuscular reflexes recorded from the lower limb in man during different tasks. AB - 1. Cutaneomuscular reflexes have been recorded in ten adult subjects from extensor digitorum brevis (EDB), tibialis anterior (TA), soleus (Sol), quadriceps femoris (Quad) and erector spinae (ES) following electrical stimulation of the digital nerves of the second toe. 2. Recordings were made while subjects were instructed to activate voluntarily the relevant muscles and also when these muscles were active posturally. 3. Reflex responses could comprise three components: an initial increase in EMG (E1), followed by a decrease (I1), followed by a second increase (E2). E1 and I1 were confined to muscles acting at the ankle and in the foot. E2 was most pronounced in EDB but also found in TA, Sol, Quad (1 subject) and ES. No responses were recorded contralateral to the stimulus. 4. E2 was significantly larger when the reflex was recorded during voluntary contraction of the muscle, rather than when the muscle was active posturally. 5. E1 and I1 components are mediated via spinal pathways. E2 requires the integrity of the dorsal columns, sensorimotor cortex and corticospinal tract (Jenner & Stephens, 1982; Rowlandson & Stephens, 1985b). The present study suggests that one or more of these supraspinal pathways is more active during voluntary contraction of lower limb muscles than when these muscles are active posturally. PMID- 7473253 TI - Skin strain patterns provide kinaesthetic information to the human central nervous system. AB - 1. We investigated the contribution of skin strain-related sensory inputs to movement perception and execution in five normal volunteers. The dorsal and palmar skin of the middle phalanx and the proximal interphalangeal (PIP) joint were manipulated to generate specific strain patterns in the proximal part of the index finger. To mask sensations directly related to this manipulation, skin and deeper tissues were blocked distal to the mid-portion of the proximal phalanx of the index finger by local anaesthesia. 2. Subjects were asked to move their normal right index finger either to mimic any perceived movements of the anaesthetized finger or to touch the tip of the insentient finger. 3. All subjects readily reproduced actual movements induced by the experimenter at the anaesthetized PIP joint. However, all subjects also generated flexion movements when the experimenter did not induce actual movement but produced deformations in the sentient proximal skin that were similar to those observed during actual PIP joint flexion. Likewise, the subjects indicated extension movement at the PIP joint when strain patterns corresponding to extension movements were induced. 4. In contrast, when the skin strain in the proximal part of the index finger was damped by a ring applied just proximal to the PIP joint within the anaesthetized skin area, both tested subjects failed to perceive PIP movements that actually took place.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7473254 TI - Nitric oxide evokes pain at nociceptors of the paravascular tissue and veins in humans. AB - 1. Nitric oxide (NO) evokes pain on intracutaneous application, apparently by exciting cutaneous nociceptors. To look for similarities in the responsiveness and sensitivity of other nociceptive systems to NO we determined pain intensity concentration relations for NO applied to paravascular tissue and veins in humans. 2. NO solutions (0.4-2.0 mM) were either injected paravascularly or perfused through a vascularly isolated hand vein segment. The subjects rated pain continuously with the help of an electronically controlled visual analog scale, which made it possible to determine both the time course (latency, duration) and the intensity of NO-evoked pain. 3. Regardless of where it was applied, at concentrations above 0.7 mM NO always evoked pain of similar time course and concentration dependence. Pain increased proportionally to the concentration of applied NO, reaching subjects' tolerance maximum at four to five times the threshold concentration. 4. Pain intensity-NO concentration relations were congruent, indicating that the respective nociceptive systems are equally sensitive to NO. 5. Our observations are consistent with the hypothesis that NO is a chemical link in peripheral nociception. PMID- 7473255 TI - Investigating the origin of cyclic changes in limb volume using mercury-in silastic strain gauge plethysmography in man. AB - 1. Vasomotion, a phenomen frequently observed in skeletal muscle microcirculation, has been observed under physiological conditions and found enhanced during critical reduction of tissue perfusion due to hypoxic hypoxia, haemorrhage and local hypotension. We used a computer assisted non-invasive plethysmographic method to investigate periodic changes of limb volume (volumotion), which we previously found in critically ill patients. The current study was designed to investigate the origin of volumotion. 2. Simultaneous recordings of limb circumference of both calves were obtained. In patients with peripheral vascular disease and patients with minor surgery the cross-correlation with spontaneous breathing was investigated. In patients who had undergone major abdominal or vascular surgery we analysed for cross-correlations between MSG plethysmography signals of both legs and changes in central venous, arterial and ventilation pressures. In this group the effects of positive-pressure ventilation and positive end-expiratory pressure (PEEP) were also studied. 3. No ventilation related volumotion was observed during spontaneous breathing. During positive pressure ventilation a ventilation-related peak was found in the Fourier transform in agreement with the ventilation frequency applied. The amplitude of ventilation-related volumotion decreased significantly after a pressure cuff applied to the thigh was inflated above central venous pressure. We observed a significant increase in the amplitude of ventilation-related volumotion when PEEP was applied. 4. Ventilation-related volumotion showed significant cross correlation between both legs. Slow wave volumotion (0.5-7 cycles min-1) was frequently observed, but always appeared unilaterally. Whilst we looked for correlations between slow wave volumotion and changes in central venous pressure and arterial blood pressure, respectively, significant cross-correlation with the MSG plethysmography recordings was only observed at the frequency of the positive pressure ventilation. 5. The fact that periodic changes of limb circumference matching the frequency of the positive-pressure ventilation were detectable with the MSG plethysmography system demonstrates that small volume changes (less than 0.02 ml (100 ml tissue)-1) can be assessed using this system. As slow wave volumotion was observed unilaterally and revealed no correlation either with breathing, ventilation or arterial and central venous pressure, we suggest that slow wave volumotion is a local event most likely reflecting arteriolar vasomotion. PMID- 7473256 TI - Role of airway receptors in the reflex responses of human inspiratory muscles to airway occlusion. AB - 1. This study was designed to investigate the role of airway receptors in the responses of a range of inspiratory muscles to airway occlusion. The occlusion had a rapid onset (< 10 ms), lasted 250 ms and produced only a slight impediment to inspiration. 2. Based on analysis of single trials and averaged rectified electromyographic responses (EMG) in six subjects, there was a major inhibition (IR) with an onset at 34 +/- 2 ms and a trough at 65 +/- 2 ms, and an excitation (ER) with a peak at 105 +/- 2 ms. These two responses are reflex given that voluntary reaction times to a tap on the chest wall occurred at latencies longer than the peak of ER. 3. The responses to airway occlusion did not appear in limb muscles which contracted phasically with inspiration. 4. Anaesthesia of the surface receptors of the upper airway did not attenuate the responses to occlusion. Because this procedure does not eliminate the inputs from muscle and deep laryngeal pressure receptors, two subjects were tested when intubated with a cuffed endotracheal tube so that the occlusion was delivered only to structures below the level of the trachea. Responses to airway occlusion were preserved when all upper airway receptors were 'bypassed'. 5. Responses to airway occlusion also remained after prolonged inhalation of nebulized lidocaine (lignocaine) sufficient to block the cough reflex. 6. The receptors mediating the responses to airway occlusion are therefore likely to reside in inspiratory muscles acting on the chest wall. If so, the short-latency inhibition contrasts with the excitatory stretch reflex responses observed in limb muscles. PMID- 7473258 TI - Calcium current reactivation after flash photolysis of nifedipine in skeletal muscle fibres of the frog. AB - 1. L-type calcium currents were activated by depolarization of cut muscle fibres of the frog. The current was blocked by the dihydropyridine compound nifedipine (5-10 microM) and reactivated by flash photolysis of the drug. 2. In the presence of nifedipine, increasing the time interval between the onset of depolarization and the flash resulted in progressively faster kinetics of the flash-induced current. This change developed with a slow time course similar to that of normal current activation. 3. A fast gating mode of the normally slow L-type channel was induced by conditioning activation (500 ms prepulses) applied 80 ms before a test step to the same potential. After block by nifedipine, flash-photolysis was carried out 40 ms before the end of the long conditioning pulse. The flash induced current had the same rapid time course as the current activated by the subsequent test voltage step. 4. Similarly, the time course of current activation was comparable for the voltage-induced fast mode activation (flash applied 5 ms before the test step) and the flash-induced activation 40 ms after the onset of the test depolarization. 5. Our data suggest that in frog skeletal muscle nifedipine inhibits calcium current activation by blocking a rapid channel gating step while the slow conformational change that normally limits the rate of activation of the L-type calcium channel remains unaffected. UV flash illumination results in a fast reactivation indicating that the channels need not be inactivated to be blocked by nifedipine. PMID- 7473257 TI - KATP channel mediation of anoxia-induced outward current in rat dorsal vagal neurons in vitro. AB - 1. Thin brainstem slices (150 microns thickness) were taken from mature rats, and membrane potentials (Em) and currents (Im) in the dorsal vagal neurons (DVN) were analysed with whole-cell patch clamp techniques during anoxia. 2. At a holding potential (Vh) of -50 mV, a sustained anoxia-induced outward current (AOC) of 92 +/- 44 pA (reversal potential (Erev), -78 +/- 12 mV) and a concomitant increase of membrane conductance (gm) from 2.2 +/- 0.45 to 5.9 +/- 2.4 nS were revealed in 40% of 142 DVN analysed. The AOC led to a hyperpolarization of the cells by 14.4 +/- 6.1 mV from a mean resting Em of -51 +/- 6 mV, and to blockade of spontaneous action potential discharges. In the remaining DVN, anoxia had almost no effect on Em, Im or gm and did not block spontaneous action potential discharges. 3. The AOC was not affected by 0.5 microM tetrodotoxin (TTX), 2 mM Mn2+, 50 microM cyanonitroquinoxaline dione (CNQX) or 100 microM bicuculline. 4. Elevation of the extracellular [K+] from 3 to 10 mM resulted in a positive shift of Erev of the AOC by 23 mV, whereas an increase in the [Cl-] of the patch pipette solution from 5 to 144 mM had no effect on Erev. 5. In DVN responding with an AOC, addition of 200 microM diazoxide, an activator of ATP-sensitive K+ (KATP) channels, to oxygenated solutions elicited a similar outward current (Erev = -79 +/- 5.5 mV, n = 12) and increase in gm. Diazoxide did not affect Em, Im or gm in cells which did not show an AOC. 6. In a subpopulation of DVN (n = 26), spontaneous activation of a KATP current with an Erev of -80 +/- 6 mV was observed. As analysed in four of these cells, an AOC was revealed during the initial phase of development of the spontaneous outward current but not under steady-state conditions. 7. The AOC, the diazoxide-induced current, and the spontaneous outward current were completely blocked upon bath application of the KATP channel blocker tolbutamide (100-200 microM). 8. The results indicate that the sustained anoxia-induced outward current of dorsal vagal neurons is due to activation of KATP channels. A possible physiological role of functional inactivation of these cells during metabolic disturbances is discussed. PMID- 7473259 TI - Phosphoarginine stimulation of Na(+)-Ca2+ exchange in squid axons--a new pathway for metabolic regulation? AB - 1. [Na+]o-dependent Ca2+ efflux (forward Na(+)-Ca2+ exchange), [32P]ATP wash-out curves and [ATP] were measured in internally dialysed squid giant axons at 17-18 degrees C. 2. We found that dialysing squid axons without ATP and with [Ca2+]i around 1 microM the basal levels of the [Na+]o-dependent Ca2+ efflux were significantly higher in the presence of N omega-phosphoarginine (PA). Phosphocreatine, a related phosphagen, is without effect. 3. PA stimulation of the Na(+)-Ca2+ exchange occurs in the complete absence of ATP (< 1 microM), being independent of, and additive to, the ATP-stimulated [Na+]o-dependent Ca2+ efflux. PA stimulation of [Na+]o-dependent Ca2+ efflux is fully and rapidly reversible with a Km around 7.7 mM. Activation by saturating [PA] is equivalent in magnitude to that of ATP. 4. PA stimulation of Na(+)-Ca2+ exchange is markedly dependent on intracellular Ca2+ and Mg2+ ions. Below 0.5 microM Ca2+i PA effect is negligible, becoming noticeable between 0.8 and 2 microM. In addition, Ca2+i considerably increases the rate at which PA activates the Na(+)-Ca2+ exchange. Although there is no absolute requirement of the PA effect for Mg2+ ions, this divalent cation largely stimulates the PA effect. 5. This work demonstrates, for the first time, the presence in squid axons of a new form of metabolic regulation of the Na(+) Ca2+ exchange directly and solely related to PA and different from that of MgATP. This novel mechanism is likely to play a physiological role in Ca2+ extrusion through the Na(+)-Ca2+ exchanger, particularly at micromolar [Ca2+]i. PMID- 7473262 TI - Rehabilitation research and development progress reports. 1994. PMID- 7473260 TI - Characterization of single voltage-gated Na+ and Ca2+ channels in apical dendrites of rat CA1 pyramidal neurons. AB - 1. We have used dendrite-attached patch-clamp techniques to record single Na+ and Ca2+ channel activity from the apical dendrites (up to 350 microns away from soma) of CA1 pyramidal neurons in rat hippocampal slices (ages: 2-8 weeks). 2. Na+ channels were found in every patch examined (range: 2 to > 20 channels per patch). Channel openings, which had a slope conductance of 15 +/- 0.3 pS (mean +/ S.E.M.), began with test commands to around -50 mV and consisted of both early transient channel activity and also later occurring prolonged openings of 5-15 ms. All Na+ channel activity was suppressed by inclusion of TTX (1 microM) in the recording pipette. 3. Ca2+ channel activity was recorded in about 80% of the patches examined (range: 1 to > 10 channels per patch). Several types of channel behaviour were observed in these patches. Single channel recordings in 110 mM BaCl2, revealed an approximately 10 pS channel of small unitary current amplitude (-0.5 pA at -20 mV). These channels began activating at relatively hyperpolarized potentials (-50 mV) and ensemble averages of this low voltage-activated (LVA) channel activity showed rapid inactivation. 4. A somewhat heterogeneous population of high voltage-activated, moderate conductance (HVAm; approximately 17 pS), Ca2+ channel activity was also encountered. These channels exhibited a relatively large unitary amplitude (-0.8 pA at 0 mV) and ensemble averages demonstrated moderate inactivation. The HVAm population of channels could be tentatively subdivided into two separate groups based upon mean channel open times. 5. Less frequently, HVA, large conductance (27 pS) Ca2+ channel activity (HVA1) was also observed. This large unitary amplitude (-1.5 pA at 0 mV) channel activity began with steps to approximately 0 mV and ensemble averages did not show any time-dependent inactivation. The dihydropyridine Ca2+ channel agonist Bay K 8644 (0.5 or 1 microM) was found to characteristically prolong these channel openings. 6. omega-Conotoxin MVIIC (10 microM), did not significantly reduce the amount of channel activity recorded from the LVA, HVAm or HVA1 channel types in dendritic patches. In patches from somata, omega-conotoxin MVIIC was effective in eliminating a significant amount of HVAm Ca2+ channel activity. Inclusion of 50 or 100 microM NiCl2 to the recording solution significantly reduced the amount of channel activity recorded from LVA and HVAm channel types in dendritic patches. A subpopulation of HVAm channels was, however, found to be Ni2+ insensitive. Dendritic HVA, channel activity was unaffected by these low concentrations of Ni2+.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 7473263 TI - Burning mouth syndrome: the relationship between the HAD scale and parafunctional habits. AB - This study investigated 84 patients with burning mouth syndrome (BMS), who were asked to complete a hospital anxiety and depression (HAD) scale questionnaire. A control group of 69 patients was also included. All patients were interviewed regarding parafunctional habits and were subjectively examined for signs of occlusal wear of the natural teeth or dentures. The results demonstrated that parafunctional habits were present in 61% of patients with BMS. There was a statistically significantly relationship between parafunctional habits and anxiety as indicated by the HAD scale, but not with depression. PMID- 7473261 TI - Single voltage-activated Na+ and K+ channels in the somata of rat motoneurones. AB - 1. Voltage-activated Na+ and K+ channels were investigated in the soma membrane of motoneurones using the patch-clamp technique applied to thin slices of neonatal rat spinal cord. 2. One type of TTX-sensitive Na+ channel, with a conductance of 14.0 pS, was found to underlie the macroscopic Na+ conductance in the somata of motoneurones. These channels activated within a potential range between -60 and -20 mV with a potential of half-maximal activation (E50) of -38.9 mV and steepness factor (k) of 6.1 mV. 3. Kinetics of Na+ channel inactivation could be fitted with a single exponential function at all potentials investigated. The curve of the steady-state inactivation had the following parameters: a half-maximal potential (Eh,50) of -81.6 mV and k of -10.2 mV. 4. Kinetics of recovery of Na+ channels from inactivation at a potential of -80 mV were double exponential with fast and slow components of 16.2 (76%) and 153.7 ms (24%), respectively. It is suggested that the recovery of Na+ channels from inactivation plays a major role in defining the limiting firing frequency of action potentials in motoneurones. 5. Whole-cell K+ currents consisted of transient (A)- and delayed-rectifier (DR)-components. The A-component activated between -60 and +20 mV with an E50 of -33.3 mV and k of 15.7 mV. The curve of steady-state inactivation was best fitted with an Eh,50 of -82.5 mV and k of 10.2 mV. The DR-component of K+ current activated smoothly at more positive potentials. E50 and k for DR-currents were +1.4 and 16.9 mV, respectively. 6. The most frequent single K+ channel found in the somata of motoneurones was the fast inactivating A-channel with a conductance of 19.2 pS in external Ringer solution. In symmetrical high-K+ solutions the conductance was 50.9 and 39.6 pS for inward and outward currents, respectively. The channel activation took place between -60 and +20 mV. The curve of steady-state inactivation of single A-channels had an Eh,50 of -87.1 mV and k of -12.8 mV. In high-Ko+ solution A-channels demonstrated a rapid deactivation at potentials between -110 and -60 mV. The time constant of the channel deactivation depended on the membrane potential and changed from 1.5 ms at -110 mV to 6.3 ms at -60 mV. 7. Delayed-rectifier K+ channels were found in the soma membrane at a moderate density.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7473264 TI - Detection of cytomegalovirus and Epstein-Barr virus in labial salivary glands in Sjogren's syndrome and non-specific sialadenitis. AB - To investigate the role of herpes viruses in Sjogren's syndrome, minor (labial) salivary gland tissues from Sjogren's syndrome and from non-specific sialadenitis were examined for Epstein-Barr virus (EBV) and human cytomegalovirus (HCMV) DNA by the polymerase chain reaction. Almost half of all salivary glands studied contained EBV and/or HCMV. There was, however, no significant difference between the detection of EBV or HCMV in salivary glands from patients with Sjogren's syndrome or non-specific sialadenitis. The findings are consistent with the persistence of EBV and HCMV in minor salivary glands following primary infection, but do not indicate a direct role for either virus in the aetiology of Sjogren's syndrome, and do not exclude reactivation of the viruses in this disease. PMID- 7473265 TI - Proteolytic enzymes in salivary extravasation mucoceles. AB - We examined the content of type IV collagenases and plasminogen activators (PAs) in luminal fluid of four extravasation mucoceles by means of gelatin or casein zymography. Immunohistochemical staining for type IV collagenases and PAs was performed to identify the possible source of these enzymes. The luminal fluid examined by gelatin zymography showed a high level of type IV collagenases compared with saliva from Wharton's duct. PAs were detected in only one of the cases by casein zymography, and none was detected in the saliva. By immunohistochemical staining, type IV collagenases and PAs were detected in duct and myoepithelial cells of the adjacent salivary gland and in macrophages and fibroblasts of the cyst wall. The results suggest that proteolytic enzymes are involved in the pathogenesis of mucoceles. PMID- 7473267 TI - Vacuolated cells and mucous metaplasia in the epithelial linings of radicular and residual cysts. AB - The purpose of this study was to determine whether a consistent association exists between mucous cells and clear or vacuolated cells in the epithelial lining of radicular and residual cysts and to consider whether the vacuolated cells may represent a stage in the histogenesis of mucous metaplasia in these linings. Single sections from each of 154 mandibular radicular and residual cysts were stained with periodic acid-Schiff (PAS) after diastase digestion. Fifteen cases which showed mucous metaplasia were included in the study and were examined for the presence of vacuolated cells associated with the mucous cells. Mucous cells were present singly or in groups within all layers of the stratified squamous epithelial lining except the basal cell layer. In nearly all instances small to large ovoid vacuolated cells were found closely associated with the mucous cells. Occasional vacuolated cells contained sparse mucin granules or a delicate network of PAS-positive, diastase-resistant material. It is suggested that the clear cells may represent a stage in the histogenesis of mucous metaplasia. PMID- 7473266 TI - Effects of testosterone on tumor induction and epidermal growth factor production in the mouse submandibular gland. AB - To determine whether testosterone administration was capable of modifying salivary gland carcinogenesis, female mice were given 1 mg of 9,10-dimethyl-1,2 benzanthracene (DMBA) into the left submandibular gland and then Group 1 mice received 5 mg of testosterone propionate and Group 2 mice received vehicle, olive oil, subcutaneously for 8 weeks. Twelve weeks after the start of the experiment, the weight of the left submandibular gland of the Group 2 mice was greater than that of the Group 1 mice. The incidences of submandibular gland carcinoma in Groups 1 and 2 were 41% (12/29) and 57% (17/30), respectively. Epidermal growth factor (EGF) levels of the left submandibular gland were significantly higher in Group 1 as compared with Group 2. These findings indicate that testosterone increases the production of EGF in the DMBA-injected submandibular gland, but does not promote the development of submandibular gland carcinoma. PMID- 7473268 TI - Proliferating cell nuclear antigen (PCNA) expression in oral squamous cell carcinomas. AB - Proliferating cell nuclear antigen (PCNA) is a nuclear protein synthesized in the late G1 and S phase of the cell cycle which can be detected immunohistochemically in paraffin-embedded tissue as a useful marker for the proliferating fraction of cells in tissue specimens. Thirty-eight cases of oral squamous cell carcinoma were studied in groups according to clinical staging and histological grading (UICC system), these included 5 cases of T1, 13 cases of T2, and 20 cases of T3, and 19 cases of G1, 12 cases of G2, and 7 cases of G3. The mean percentage of PCNA positively stained tumor cells in this series was 29.2. The mean percentages of PCNA-positive cells in each stage and grade were: T1 (13.7%), T2 (24.4%), and T3 (34.3%); G1 (30.5%), G2 (24.8%), and G3 (33.2%). The results indicate that the percentage of PCNA positively stained tumor cells in oral squamous cell carcinoma was significantly greater when tumor size was larger than 2 cm (T1 versus T2/T3). However, there was no significant difference between T2 and T3. There was no correlation between the number of PCNA-stained tumor cells and histological grade. PMID- 7473269 TI - Immunohistochemical demonstration of epithelial glutathione S-transferase isoenzymes in normal, benign, premalignant and malignant human oral mucosa. AB - The expression and localization of glutathione S-transferase (GST) isoenzymes in the epithelium of normal oral mucosa (n = 9), overlying reactive fibrous hyperplasia (n = 9), and of potentially malignant [leukoplakia (n = 25), submucous fibrosis (n = 12), verrucous hyperplasia (n = 16)] and malignant [squamous cell carcinoma (n = 36), verrucous carcinoma (n = 13)] oral lesions were examined immunohistochemically using polyclonal antibodies raised against GST isoenzymes (alpha, mu and pi) with the standard avidin-biotin-peroxidase complex (ABC) method. GST alpha, mu and pi were almost completely absent in the epithelium of normal oral mucosa and overlying benign fibrous tissues. GST alpha staining was cytoplasmic and focally positive, while GST mu staining was similar to but weaker than that seen for GST alpha. GST pi showed both cytoplasmic and nuclear staining and was expressed in 60% of leukoplakias with mild dysplasia (n = 15), 80% of leukoplakias with moderate to severe dysplasia (n = 10). 75% of submucous fibrosis samples (n = 12), 75% of verrucous hyperplasias (n = 16), 77% of verrucous carcinomas (n = 13), 81% of well-differentiated squamous cell carcinomas (n = 26) and 70% of moderate- to poorly-differentiated squamous cell carcinomas (n = 10). In addition, GST pi expression was independent of the state of differentiation of oral cancers. Since GST pi was significantly over-expressed in the oral premalignant and malignant lesions, the kinetics of GST pi-positive cells and the value of GST pi as a tumor marker in oral carcinogenesis need further investigation. PMID- 7473270 TI - Immunocytochemical detection of ornithine decarboxylase (ODC) in paraffin embedded tissues as a possible prognostic indicator for oral lesions. AB - Intracellular levels of ornithine decarboxylase (ODC) are raised following mitogenic stimulus and in neoplasia. Because lesions of the oral cavity are often difficult to assess histologically, we have determined the value of immunocytochemical detection of ODC as a prognostic indicator in 74 routinely fixed and paraffin-embedded oral biopsies using peroxidase-antiperoxidase and immunogold-silver staining. The latter proved more sensitive, yielding positive reactions in 32 of 43 oral carcinomas (11/14 well differentiated, 16/21 moderately differentiated and 3/5 poorly differentiated) and 7/11 potentially malignant lesions, compared with 19/45 carcinomas and 1/15 potentially malignant lesions, by peroxidase anti-peroxidase. Hyperplastic lesions (n = 7) and normal non-keratinized buccal mucosa (n = 7) were all negative. Follow-up was possible in 13 of the carcinoma patients. Of 7 positive ODC reactions but clinically node free at biopsy, 2 died and 2 had recurrences within 3 years, whereas all of 6 with no immunoreactivity were symptom-free after 3-5 years. Immunostaining for ODC may be helpful for the prognostic assessment of routinely processed oral lesions and in choosing treatment. PMID- 7473271 TI - A polymerase chain reaction (PCR) investigation of oral verrucae which contain HPV types 2 and 57 by in situ hybridization. AB - The polymerase chain reaction (PCR) and direct DNA sequencing have been used to identify strain variants of HPV types 2a/57 in formalin-fixed sections of human oral verrucae, where the virus had previously been detected by both immunofluorescence and in situ hybridization. By employing type-specific and type common PCR primers we show that these lesions contain a mixture of viral DNAs which vary by up to 27% in DNA sequence, in a region where the variation between HPV types 2a and 57 is only 4%. The extra discriminatory power of fluorescent sequencing indicates that the lesions may also contain wild-type HPV2a/57 DNA which could provide a helper function for defective viral DNA molecules or indicate a mosaic origin for the lesions. PMID- 7473272 TI - [Optical measurement of intracellular calcium concentration]. PMID- 7473274 TI - The accuracy of a high-precision digitizer for CAD/CAM of crowns. AB - An alternative method for manufacturing fixed prostheses that uses machine digitalization and a combination of milling and spark erosion has been developed. The digitizer used has a contact probe with a ball-shaped tip. This study evaluated the accuracy of the device and the transfer of data from the individual die to the computer. The dimensions of a square gauge as read by the machine were compared with the known dimensions of the object with a computer, which allowed determination of the accuracy of the digitalization. Procera machining and the implications for reliable prosthodontic treatment are discussed. PMID- 7473273 TI - Secondary caries around amalgam restorations. AB - Secondary caries is one of the greatest causes for replacement of amalgam restorations. This study verified whether marginal defects in class I restorations could be a determining factor in the development of secondary caries. Fifty-five extracted premolars and molars whose occlusal surfaces were already restored with amalgam were selected. A macroscopic examination of these teeth was carried out to see whether there was a difference in the prevalence of caries beneath ditched and nonditched margins. To assess the presence of caries, a section was ground in each tooth so that the cut passed through a ditched margin and a better margin. This permitted assessment of the two types of margins that had been exposed to the same oral environment. Macroscopic examination revealed caries in 3.6% of the nonditched margins and caries in 20% of the ditched margins. Microscopic examination revealed caries in 47.06% of the nonditched margins and caries in 58.82% of the ditched margins. Statistical analysis supports the conclusion that the marginal defect itself is insufficient to determine the presence of secondary caries that surround the amalgam restoration on the occlusal surface. PMID- 7473275 TI - Tensile bond strength of a composite resin cement for bonded prosthesis to various dental alloys. AB - The development of composite resin cements that chemically bond to dental alloys has improved the construction of resin-bonded prostheses. Composite resins can be selected for various situations, but specific clinical situations may require different alloys. This study evaluated the ability of a composite resin cement to bond to various dental alloys of different compositions. Ten pairs of disks for each alloy (two NiCr, two NiCrBe, one CuAl, one gold type IV, and one gold for metal ceramic) were bonded to a composite resin cement after air abrasion was performed with aluminum oxide. The disks were then rinsed in tap water and were ultrasonically cleaned in distilled water for 2 minutes. The tensile tests exhibited greater values for alloys ultrasonically cleaned, and the best results were recorded by NiCr and NiCrBe alloys. PMID- 7473276 TI - O-ring coping attachments for removable partial dentures. AB - Clinical experiences for the past 9 years have demonstrated the practicability of the O-ring coping attachment, a modification of the telescopic crown-and-sleeve coping retainers, for removable partial dentures. A circumferential groove placed in the primary coping receives an elastomeric O-ring that fits into a corresponding groove made in the internal surface of the telescopic crown. The O ring not only provides controllable retention but also acts as a shock absorber. Long-term retention of the prostheses can be easily maintained by periodic replacements of the O-rings. Sophisticated procedures and expensive machines are not required to make the prostheses. Excellent patient acceptance and the versatility in clinical applications make this system one of the winning designs for removable partial prosthodontics. PMID- 7473277 TI - Adhesion of denture tooth porcelain to heat-polymerized denture resin. AB - Use of porcelain denture teeth may be desirable in many clinical situations, including implant-supported prostheses. However, lack of space because of frameworks often precludes the use of conventional retention by diatorics and pins. Adhesion of porcelain denture teeth to denture resin could also stiffen and possibly strengthen dentures and decrease stain ingress between porcelain teeth and resin denture bases. Unlike previous studies that investigated the bond between conventional feldspathic metal-ceramic porcelain and bis-GMA based composite resin, this study investigated adhesion of denture tooth porcelain to polymethyl methacrylate (PMMA). High-energy air abrasion, hydrofluoric acid etching, and the use of a general purpose bonding agent resulted in an improved bond strength of heat-cured denture PMMA bonded to denture tooth porcelain. Silane coating did not improve bond strengths, and conventional air abrasion was no more effective than polishing with 600-grit silicon carbide. Storage in water and artificial aging substantially decreased bond strengths. The strongest bond strengths were achieved by a high-energy-abrasion + etching + multi-purpose bonding-agent treatment, but a simpler etching + multiple-purpose bonding-agent treatment also produced reliable results. A laboratory technique was suggested. The role of surface treatment in the mechanism of adhesion was examined with scanning electron microscopy. High-energy abrasion produced a slightly more detailed initial topography than conventional air abrasion, but after etching, the high-energy topography became much more detailed. Surface topography alone did not account for all differences found. PMID- 7473279 TI - Measurement (in vitro) of the amount of force required to dislodge specific clasps from different depths of undercut. AB - A fundamental principle in clasp selection for a specific abutment is the reduction of the transmission of excessive forces to that abutment. The flexibility of a clasp was believed to directly affect the reduction of such forces. This study compares the average measurements of forces required to dislodge two kinds of circumferential clasps in different amounts of undercuts: one with a half-round retentive arm and the other with a round retentive arm under tensile load. Three commonly used undercuts were created on six cast crowns, premolars, and molars. Each clasp was exposed to adequate tensile load until dislodgment occurred. The test was run six times for the same clasp. The means of tensile load required to dislodge each of the different clasps were compared statistically. However, no significant difference was detected for the main effects of the three variables studied: tooth type, undercut, and amount and clasp type. Findings indicated the possible use of cast round clasps where advantages of clinical fit and reduction of transmitted forces to the abutments can be gained. PMID- 7473278 TI - Mechanical properties of laser-welded cast and wrought titanium. AB - This study evaluated the mechanical properties of laser-welded cast and wrought titanium base and compared them with those of a brazed type IV casting gold alloy. Ultimate tensile strength, 0.2% yield strength, and percent elongation were recorded for joined and unjoined bars of the previously mentioned materials. Sections of titanium bars were laser-welded, and gold alloy bars were brazed. Both joining methods significantly reduced the ductility of the material. The strength of the cast gold alloy was superior to that of titanium. However, the strength of the laser-welded titanium equaled that of the brazed gold alloy, which suggests that dental restorations made of cast and wrought titanium would satisfy ordinary clinical requirements. PMID- 7473281 TI - Comparison of screw loosening, rotation, and deflection among three implant designs. AB - A common problem associated with single tooth implant restorations is abutment screw loosening. Manufacturers of implants have attempted to overcome this problem by incorporating antirotational design characteristics into their systems. Micromovement and torque levels required to loosen abutment screws for straight and angled antirotational screw-retained abutment/implant combinations from three different manufacturers were examined in this in vitro investigation. A custom-built machine was used and each sample was subjected to compressive horizontal reciprocal movements over a 25-degree incline for a simulated 1-month period. Data were generated that showed movements of the crown/abutment complex during force application. The amount of torque necessary to loosen the abutment screws before and after testing was also recorded and compared for each system. The results indicated no significant differences (p < 0.05) among all the straight and angled abutments for the variables studied. PMID- 7473280 TI - Denture replacement during a 14-year period in Alberta's universal dental plan for the elderly. AB - Since 1973 Alberta's dental plan for the elderly has made government-sponsored, premium-free, comprehensive dental care available from both dentists and denturists in private practice to all residents older than 64 years. This article is based on an analysis that covered 260,000 patients from 1978 to 1992. It presents the frequency, nature, and cost of denture replacements to patients and pays particular attention to replacements administered within the allowable 5 year time limit stated by the plan for complete and removable partial dentures. During the 14-year period, about 55,000 dentures were replaced; 47% of these were replaced in the year after the individual's 5-year time limit had expired. Within the 5-year limit 1974 dentures were replaced at a nominal cost of +1.09 million, with nearly one half of these costs being attributed to denturists. Relative to the total number of dentures provided during the 14-year period, the denture replacement rate for dentists was 7% to 8%, and for denturists it was nearly three times greater at 21% to 22%. The replacement process included many crossovers by patients between dentists and denturists; for example, nearly one quarter of the 190 complete dentures initially provided by dentists were replaced by denturists within 5 years. PMID- 7473282 TI - Computer simulation of occlusal discrepancies resulting from different mounting techniques. AB - The effect of arbitrary mounting of maxillary casts on occlusal relationships was investigated in this study. Maxillary casts of 31 volunteers were mounted on an articulator by use of two split cast bases. This mounting was done first with the arbitrary face bow and second with a hinge bow. Three reference points were defined and measured on each maxillary cast with a three-dimensional digitizer. The measurements were taken from the arbitrarily mounted cast and from the cast mounted according to the hinge axis. Opening and closing movements that were transferred according to the hinge axis. Opening and closing movements that were transferred from the articulator to the mouth of the patient were simulated by a computer based on measurements of the reference points. The results revealed that the use of an arbitrary face bow causes a deviation of the hinge-axis points from the precise axis of more than 5 mm in 77% of the cases. Resulting occlusal errors depended on the angles between the arbitrary and precise axes and the direction of the axis shifts. The occlusal error is roughly proportional to the shift or tilting of the hinge axis in millimeters or degrees. For a given deviation of the arbitrary and precise axes, the occlusal error is proportional to the record height. For a record height of 2 mm or more, an occlusal error of more than 0.1 mm will occur. An average occlusal error of more than 0.1 mm would most likely lead to the necessity of extensive selective grinding of occlusal discrepancies in the patient's mouth. PMID- 7473283 TI - Stereo laser-welded titanium implant frameworks: clinical and laboratory procedures with a summary of 1-year clinical trials. AB - A new technology for implant framework fabrication (Procera) has been introduced to North America after initial clinical trials in Sweden. This technique is unique because it eliminates the conventional approach to framework fabrication with the lost wax casting technique. Clinical and laboratory procedures associated with the Procera technique are described in this article, as are the most recent developments in this rapidly emerging technology. A report of 1-year results of the first 10 patients treated with the Procera technique at the University of Washington is described. Treatment outcome has been favorable, with no clinical evidence of prosthodonic or soft-tissue complications noted. All 50 of the implants placed in the 10 patients remain integrated after 1 year of service. The Procera technology offers an alternative to current conventional framework fabrication techniques. Continued follow-up of this patient series is pursued to explore the treatment outcome for a longer interval of time. PMID- 7473284 TI - Release of asbestos fibers during casting ring liner manipulation. AB - Civil lawsuits have been filed that allege wrongful deaths of at least one dentist and one dental technician from asbestos exposure. However, no known published scientific studies support such allegations. To assess what potential exposure could occur, air samples were collected while casting rings were lined with asbestos liner. Fibers were collected on filters and counted by use of phase contrast microscopy and transmission electron microscopy. The concentrations of fibers 5 microns long or greater on all filters was less than allowed by federal guidelines. Based on this simulation the potential health risk from using asbestos ring liner seems low. PMID- 7473285 TI - Dynamic viscoelastic properties of processed soft denture liners: Part II--Effect of aging. AB - The proper functioning of soft denture liners depends to a great extent on their mechanical properties. As with many polymers these materials are affected by aging. Twelve soft denture liners were processed by a laboratory according to the manufacturers' directions. Five specimens of each material were tested without aging. Five additional specimens of each material were subjected to 900 hours of accelerated aging in a Weather-Ometer instrument. These were tested with a dynamic viscoelastometer at three frequencies and two temperatures, and data for 37 degrees C and 1 Hz was obtained. Two of the ethyl methacrylate resins demonstrated the largest increases in storage (E') and loss moduli (E") after aging. These materials also showed the greatest overall E' and E". One denture liner material exhibited 673% and 488% increases in E' and E", and other materials showed smaller increases. The effects of aging on the damping factor (tan delta) were varied and five materials showed increased tan delta. Only two ethyl methacrylate resins developed lower tan delta. All the silicone and polyphosphazine rubbers showed small changes after aging and had the lowest tan delta values. Significance of differences between materials and treatments was tested with ANOVA, Scheffe intervals, and t-tests at a = 0.05. The ethyl methacrylate soft denture liners were affected the most by accelerated aging, and the silicones and polyphosphazine were least affected. The ethyl methacrylate resins also had the greatest values of E', E", and tan delta after aging. PMID- 7473286 TI - Copper band matrix for large Class V amalgam restorations. AB - Class V cavities present special problems in restoration and maintenance. The subsequent health of the gingival tissue and the relative immunity toward recurrent caries depends not only on the age of the patient and patient's oral hygiene but to a great extent on the excellence of the restoration. This excellence is achieved by the operator's detailed design of the preparation, moisture control, and restoration and material finishing. Dental amalgam continues to be the material of choice in many clinical situations. The restoration of a large class V lesion that results from replacement of a defective restoration or extensive caries benefits greatly when a copper band matrix is used because of its ability to be shaped and contoured and its rigidity. In large class V cavities, the copper band creates a "false" wall that is required for confinement of the amalgam as it must be packed under pressure. This confinement of the cavity is necessary because it restricts the amount of excess material which ultimately is important in the restoration of the tooth and its long-term success. PMID- 7473287 TI - In-depth disinfection of acrylic resins. AB - This study demonstrated that bacteria penetrate three kinds of dental acrylic resin after a short time period. Samples of acrylic resin were contaminated with a variety of bacteria and were then placed in three different disinfecting solutions as directed by the manufacturers. After the specific dilution and immersion time, cultures were made from the resin samples. The only effective disinfectant was a 0.525% solution of sodium hypochlorite at a 10-minute immersion. It disinfected not only the surfaces but also the bacteria that penetrated the surfaces to a depth of 3 mm. PMID- 7473288 TI - Simplified one-step procedure for making impressions and jaw relation records of implant-supported reconstruction. PMID- 7473289 TI - Use of acrylic resin copings as adjuncts for groove placement during tooth preparation. PMID- 7473291 TI - Esthetic consideration of a labial bar major connector. PMID- 7473292 TI - Possible acceleration of age effects on cognition following menopause. AB - Many cognitive functions have been shown to deteriorate with age. Because of the importance of the menopause as a milestone in the life cycles of women, we examined whether the aging-over-time process in some cognitive functions differs between women of reproductive age and postmenopausal women. It is demonstrated here that in some cognitive tests, including driving simulation, reaction time and some visuospatial tests, there is a significant acceleration in deterioration of functioning following menopause. It is suggested that this acceleration might be associated with the lack of gonadal hormones or other reproduction-related factors which may play a protective role against age-related deterioration in some cognitive functions in women. PMID- 7473290 TI - A fast and efficient method for cast core provisional restoration of anterior teeth. PMID- 7473293 TI - Relapse and recurrence following a course of ECT: reasons for concern and strategies for further investigation. AB - Patients treated with electroconvulsive therapy (ECT) have a more severe illness. This severity is evidenced in the clinical profile of the illness, in the course of the illness, in the resistance to pharmacotherapy, and in the high relapse and recurrence rates after the course of ECT. In this article the authors explore some of the issues, particularly those of biological correlates of severe mood disorders and the continuation of treatment after ECT, which may be factors in this increased severity. The authors propose avenues for the investigation and treatment of severe mood disorders treated with ECT. PMID- 7473294 TI - Hemispheric asymmetries of function in patients with major affective disorders. AB - Several investigators have suggested that affectively disordered patients have dysfunction of the nondominant cerebral hemisphere. The present study tested this hypothesis using psychometrically matched measures of verbal and visual-spatial skills. A sample of 64 psychiatric in-patients and out-patients was interviewed using an expanded and modified version of the NIMH Diagnostic Interview Schedule and diagnosed using the criteria of DSM-III-R, the Research Diagnostic Criteria, and the Feighner et al. system. Patients diagnosed as affectively disordered by at least one system were administered a pair of psychometrically matched measures: a measure of Word-Finding modeled after the Boston Naming Test, and a measure of visual-spatial functioning adapted from the Dot-Localization task. Patients with diagnoses of major depression by any of the three systems showed significantly poorer performance on Dot Localization than on Word Finding. Differences for patients with bipolar diagnoses were in the same direction but fell short of significance. These results support the hypothesis that patients with major depressive disorders may show impaired right-hemisphere functioning. PMID- 7473295 TI - The Neurobehavioral Rating Scale: reliability in patients with dementia. AB - Precise measurement of cognitive, psychiatric, and behavioral symptoms is essential to understanding clinical, pathophysiologic, and treatment aspects of Alzheimer's disease and other dementing illnesses. The Neurobehavioral Rating Scale (NRS) is a 28-item observer-rated instrument that measures a broad range of cognitive and noncognitive symptoms. The interrater reliability of the NRS was examined in 15 patients with dementia. The correlation coefficient for the NRS total scores was .93. Coefficients for NRS factor scores and individual item scores were also satisfactory. Correlations for measures of subjectively experienced symptoms were acceptable, but less robust than measures of cognition and observable behavior. These results support the reliability of the NRS for multidimensional assessment of patients with dementia. PMID- 7473296 TI - Preferred pathways of behavioral contagion. AB - A behavioral disorder is "contagious" if the risk to a given individual increases when someone in that person's vicinity, family, or social group develops the disorder. So understood, behavioral contagion may be involved in criminality, conduct disorder, drug abuse, suicide, and teenage pregnancy. Previous papers have shown that contagion generates highly distinctive result patterns, from which its presence may be inferred. The patterns concern prevalence in sibships of different size and, in case-control designs, the number of susceptible sibs that affected and unaffected individuals have. The present paper extends the analysis by allowing the likelihood of transmission from one sib to another to vary according as the two sibs are of the same or opposite gender, male or female, single borns or co-twins, fraternal or identical twins. The extension is illustrated by application to data on criminality in Danish twins previously reported by other workers. We will show that the distribution of criminality by gender and zygosity is better explained in terms of behavioral contagion than by previous analyses. PMID- 7473297 TI - The severity of dieting and bingeing behaviors in college women: interview validation of survey data. AB - A combined survey and interview study was conducted to validate a categorical Dieting and Bingeing Severity Scale (DBSS), and to estimate the prevalence of eating disorders in young women. We hypothesized that assignment to the DBSS categories would be confirmed by clinical interviews such that interview diagnosed eating disorders would be found with increasing frequency and severity at the upper end of the DBSS. Freshmen college women (n = 1367) completed a survey instrument addressing the frequency and severity of dieting, binge-eating, and other behaviors and attitudes related to weight control. Random stratified sampling procedures were used to select a subset of women (n = 306) from each DBSS category for structured clinical interviews for DSM-III-R (SCID). Survey respondents were assigned to one of six mutually exclusive DBSS categories: non dieters (9% of sample), casual dieters (26%), moderate dieters (23%), intense dieters (21%), dieters at-risk (19%), and probable bulimia nervosa (2%). The DBSS effectively rank-ordered subjects according to the risk of having interview diagnosed eating disorders. Women in the three most severe DBSS categories were significantly more likely to have current subthreshold and threshold level eating disorders, in particular bulimia nervosa and eating disorder not otherwise specified (EDNOS). The estimated prevalence of current bulimia nervosa was approximately 2% by both survey and interview methods. The prevalence of current EDNOS was 13%, more than six times greater than the prevalence of bulimia nervosa. The DBSS was found to be a reliable and valid measure of dieting and bingeing severity. The survey instrument may be useful in measuring the extent of, and changes in, pathological dieting in community-based samples of young women, and in studying comorbidity of dieting and bingeing severity with other psychiatric conditions including depression and substance use. The DBSS may also be useful in identifying risk factors associated with the onset of eating disorders. PMID- 7473298 TI - Depleted red cell membrane essential fatty acids in drug-treated schizophrenic patients. AB - The fatty acid composition of red blood cell membranes from 23 drug-treated schizophrenic patients was measured and compared with a healthy control group. There were substantial depletions of fatty acids from the n6 and n3 series, particularly arachidonic and docosahexanoic acid. Significant negative correlations between depleted n6 fatty acids and plasma levels of thiobarbituric acid reactive substances suggests that depletion is caused by increased breakdown of these fatty acids rather than by impaired incorporation of fatty acids into membranes. Arachidonic and docosahexanoic acids appear to show a bimodal distribution. We propose that this may be a metabolic abnormality which is of aetiological importance in schizophrenia. PMID- 7473299 TI - Cerebrospinal fluid levels of monoamine metabolites and gamma-aminobutyric acid in neuroleptic malignant syndrome. AB - To investigate the pathophysiology of neuroleptic malignant syndrome (NMS), we measured various cerebrospinal fluid (CSF) parameters in 11 patients and compared them with 8 age-matched normal controls. Concentrations of homovanillic acid (HVA) were significantly decreased during the active phase of NMS. This finding indicates that the dopaminergic nervous system of the patients was in a state of hypofunction in this phase and supports the central dopamine blockade theory of NMS. Reduced CSF HVA levels were also found after recovery from NMS, suggesting that hypofunction of the dopaminergic system may continue subclinically. The levels of 5-HIAA were decreased in the active phase, but the change was not significant. Therefore, a relationship between the development of NMS and disturbances of serotonin metabolism remains unclear. The levels of noradrenaline and its major metabolite, 3-methoxy-4-hydroxyphenylethyleneglycol increased significantly during the active phase, but returned to normal after recovery. These results show the existence of sympathetic nervous system hyperactivity during the active phase of NMS. The levels of gamma-aminobutyric acid (GABA) were significantly lower in the patients than in the controls. These findings suggest that there is a GABAergic deficiency in NMS. Thus, our study indicates that, in addition to dopamine hypoactivity, disturbances in various neurotransmitter systems are involved in the pathophysiology of NMS. PMID- 7473301 TI - Chaotic analysis in psychology and psychoanalysis. AB - The rise of chaos theory in physics has been accompanied by attempts to use chaos as an explanatory tool in psychology. A review of how chaos theory is used in psychology reveals two relatively distinct efforts: chaos as a mathematical model of psychological phenomena and chaos as a metaphor for psychological phenomena. A discussion of recent articles reveals that most chaotic analysis fails to respect the minimum qualifications for data subjected to such analysis. Further, uses of chaos as an analogy for psychological phenomena are rife with misunderstandings of chaos. PMID- 7473300 TI - Lack of effect of short-term fasting on cognitive function. AB - In a study designed to assess the effects of short-term food deprivation on cognitive function, a sample of female subjects (N = 21) was tested on a number of measures of cognitive function after three levels of food deprivation (miss one meal, miss two meals or miss all food for 24 h prior to testing) and a condition in which they ate normally for 24 h prior to testing. There was found to be no significant effects of food deprivation on sustained attention, attentional focus, simple reaction time or immediate memory. However, performance on a low processing load tapping task was significantly poorer when the subjects were deprived of food for 24 h prior to testing, and heart rate was significantly higher when they were non-deprived. These results stand in contrast to the impairments in cognitive function previously found to be associated with spontaneous dieting behaviour (using essentially the same task battery). PMID- 7473303 TI - Assessing filial maturity through the use of the Filial Anxiety Scale. AB - This study was an investigation of a late-in-life stage of development known as filial maturity (Blenkner, 1965). The Filial Anxiety Scale (Cicirelli, 1988) was used in a cross-sectional design to assess whether the filial anxiety of adult caregivers of aging parents varied in a manner consistent with Blenkner's concept. Results clearly supported Blenkner's position that there is a late-in life period of development during which some individuals experience heightened levels of filial anxiety, or a filial crisis, prompted by the caregiving demands of an aging parent. This crisis is then followed by a significant decrease in anxiety and results in the attainment of a developmental period characterized by filial maturity. PMID- 7473302 TI - The beliefs of members of the American Psychiatric Association on the etiology of male homosexuality: a national survey. AB - This study is an analysis of psychiatrists' (N = 508) attitudes with regard to the etiology of male homosexuality. A random sample of members of the American Psychiatric Association revealed an endorsement of a biological perspective. Attitudes were further analyzed with regard to variation by school of thought, practice setting, and number of years practicing psychiatry. PMID- 7473304 TI - Relationships among three general approaches to personality description. AB - The present study was designed to investigate relationships among three basic models used in research on personality description: Wiggins's (1979) Circumplex, Goldberg's (1992) Big-Five, and Mehrabian's (1978, 1980) PAD Temperament models. Wiggins's Nurturance and Dominance scales were highly reliable and nearly independent, as were Mehrabian's Trait Pleasure, Trait Arousability, and Trait Dominance scales. The Big-Five scales exhibited lower reliabilities and an average interscale correlation of .21 for the Opaque and .39 for the Transparent scales. Regression and factor analyses showed that basic factors of pleasantness (Goldberg's Agreeableness, Wiggins's Nurturance, and Mehrabian's Trait Pleasure), dominance (Wiggins's Dominance and Mehrabian's Trait Dominance), and arousability (Mehrabian's Trait Arousability) provided a sufficient and reasonably parsimonious description of the variance in all the scales investigated. PMID- 7473305 TI - Visual pigments and spectral sensitivity of the diurnal gecko Gonatodes albogularis. AB - The visual pigments and oil droplets in the retina of the diurnal gecko Gonatodes albogularis were examined microspectrophotometrically, and the spectral sensitivity under various adapting conditions was recorded using electrophysiological responses. Three classes of visual pigments were identified, with lambda max at about 542, 475, and 362 nm. Spectral sensitivity functions revealed a broad range of sensitivity, with a peak at approximately 530-540 nm. The cornea and oil droplets were found to be transparent across a range from 350 700 nm, but the lens absorbed short wavelength light below 450 nm. Despite the filtering effect of the lens, a secondary peak in spectral sensitivity to ultraviolet wavelengths was found. These results suggest that G. albogularis does possess the visual mechanisms for discrimination of the color pattern of conspecifics based on either hue or brightness. These findings are discussed in terms of the variation in coloration and social behavior of Gonatodes. PMID- 7473306 TI - Postmetamorphic changes in auditory sensitivity of the bullfrog midbrain. AB - During metamorphosis, the lateral line system of ranid frogs (Rana catesbeiana) degenerates and an auditory system sensitive to airborne sounds develops. We examined the onset of function and developmental changes in the central auditory system by recording multi-unit activity from the principal nucleus of the torus semicircularis (TSp) of bullfrogs at different postmetamorphic stages in response to tympanically-presented auditory stimuli. No responses were recorded to stimuli of up to 95 dB SPL from late-metamorphic tadpoles, but auditory responses were recorded within 24 hours of completion of metamorphosis. Audiograms from froglets (SVL < 5.5 cm) were relatively flat in shape with high thresholds, and showed a decrease in most sensitive frequency (MSF) from about 2500 Hz to about 1500 Hz throughout the first 7-10 days after completion of metamorphosis. Audiograms from frogs larger than 5.5 cm showed continuous downward shifts in MSF and thresholds, and increases in sharpness around MSF until reaching adult-like values. Spontaneous activity in the TSp increased throughout postmetamorphic development. The torus increased in volume by approximately 50% throughout development and displayed changes in cell density and nuclear organization. These observations suggest that the onset of sensitivity to tympanically presented airborne sounds is limited by peripheral, rather than central, auditory maturation. PMID- 7473307 TI - Termination of leech swimming activity by a previously identified swim trigger neuron. AB - Cell Tr2 is a neuron in the subesophageal ganglion of the leech that can trigger swim episodes. In this report, we describe the ability of Tr2 to terminate ongoing swim episodes as well as to trigger swimming. Stimulation of Tr2 terminated ongoing swim episodes in nearly every preparation tested, while Tr2 stimulation triggered swim episodes in only a minority of the preparations. We suggest that the primary role of Tr2 is in the termination rather than the initiation of swimming activity. The swim trigger neuron Tr3 and a swim-gating neuron, cell 21, hyperpolarized during Tr2-induced swim termination. Another swim gating neuron, cell 204 was sometimes slightly excited, but more often, hyperpolarized during Tr2-induced swim termination. In contrast to these cells, Tr2 stimulation excited another swim-gating neuron, cell 61. The responses of the swim-gating cells were variable in amplitude and sometimes not evident during Tr2 induced swim termination. Hence, the effects of Tr2 stimulation on swim-gating neurons seem unlikely to be the direct cause of swim termination. Oscillator cells examined during Tr2-induced swim termination include: 27, 28, 33, 60, 115, and 208. The largest effect seen in an oscillator neuron was in cell 208, which was repolarized by up to 10 mV during Tr2 stimulation. Tr2 stimulation did not produce any obvious synaptic effects in motor neurons DI-1, VI-1, and DE-3. Our findings indicate that other, yet undiscovered, connections are likely to be important in Tr2-induced swim termination. Therefore, we propose that cell Tr2 is probably a member of a distributed neural network involved in swim termination. PMID- 7473308 TI - Serving time: psychiatric mental health nursing in corrections. AB - 1. The opposing philosophies of custody and caring result in direct implications for nurses working in the correctional environment. Nurses constantly "walk in line" between the requirement of security, health care, and client advocacy. 2. Deinstitutionalization, increased rates of incarceration for interpersonal violence, and substance abuse related crimes have resulted in a rapidly expanding prison population. Of all offenders, 10% to 15% have a major mental illness, leading to many treatment challenges for the nurse. 3. Therapy issues with this population are complicated. Correctional nurses must have a good understanding of the therapeutic nurse-client relationship, with particular attention to therapeutic impasses, transference, and countertransference. 4. Historically, role development for nurses working in corrections has been particularly difficult. Correctional nurses must grapple with issues surrounding professional identity, professional isolation, and territoriality. PMID- 7473309 TI - Geropsychiatric public sector nursing: placement challenges. AB - 1. The nursing home eligible resident, who has experienced long periods of institutional living in state hospitals and many unsuccessful placements in nursing home facilities, presents a challenge in developing a viable community treatment plan. 2. Mental health consultations with nursing home staff are valuable resources for developing and implementing additional behavioral management interventions. 3. Collaborative efforts of public sector health care agencies have resulted in a cost effective approach to decreasing hospital admissions and recidivism and increasing community tenure of geropsychiatric residents. PMID- 7473310 TI - Jailed mothers: further explorations in public sector nursing. AB - 1. The burgeoning population of women, particularly pregnant women, being incarcerated increases the need for psychosocial and maternal-child nurses employed in prisons. 2. The potential paradox of incarcerated women receiving better care in prisons than when free suggests the need for public sector psychosocial nurses to address issues of social justice. 3. The great majority of informants enthusiastically responded to the idea of a mother-infant corrections residential program. They suggested that more attention be given to prenatal care and preparation for labor and delivery than pregnant women historically received in prisons. PMID- 7473311 TI - An integrated cooperation model for long-term mentally ill in the community. AB - 1. This article describes a Swedish model of community support for patients with a long-term mental illness. 2. The result confirms that the staff-psychiatric practical nurses and aids are important in the care with support and coping as essential elements. 3. The staff members do require supervision from a nurse with knowledge about society and psychiatric care. PMID- 7473312 TI - Cost containment detrimentally affects patient care. PMID- 7473313 TI - Seven days and counting: how inpatient nurses might adjust their practice to brief hospitalization. AB - 1. Inpatient nurses can help create a seamless health care system by designing groups and specific interventions that begin at the time of admission and continue after discharge. 2. Inpatient nurses must clearly articulate how they intervene for the benefit of patients and how they engineer the coordinated staff effort that maintains the basic safety and structure of the environment. 3. Nurses should learn how to organize and report observations so they formulate a concise assessment summary while also collecting data that are useful to the unit's outcome evaluation. PMID- 7473314 TI - Public sector psychosocial nursing. PMID- 7473315 TI - The transition from victim to victor: application of the theory of mastery. AB - 1. Serious, chronic disease, such as sarcoidosis, evokes life-changes that may require mastering stress, recovering self-esteem and finding alternative sources of satisfaction for what is lost. 2. The theory of mastery offers patients and nurses an explanation of how to emerge ever stronger and more resilient in dealing with the self and the environment. 3. Over time, four stages of the theory of mastery may be encountered: certainty, change, acceptance, and growth. Nurses may assist patients to see how they may "master" each stage to achieve personal growth, hence a greater quality of life. PMID- 7473316 TI - Lego therapy. PMID- 7473317 TI - Changing populations in the public sector: responding to the needs of Texas youth. AB - Treatment of sexually abused and abusing youth requires careful program planning and implementation. Led by its Quality Improvement Council and with considerable staff involvement, Waco Center for Youth in Waco, Texas developed a dual-track approach to treatment of this underserved, often misunderstood, adolescent client population. The sexual abuse survivors' program focuses on the issues of trust, safety, self-esteem, assertiveness, and education. The sex offender program is built on the concepts of self-responsibility; diminishing cognitive distortions; identification of stressors, and their relationship to the offending cycle; and relapse prevention. The staff of Waco Center for Youth are engaged in ongoing analysis of behavioral indicators for successful treatment of sexually abused and abusing adolescents. PMID- 7473318 TI - Alcohol and the heart in perspective. Sensible limits reaffirmed. A Working Group of the Royal Colleges of Physicians, Psychiatrists and General Practitioners. PMID- 7473319 TI - Incontinence. Causes, management and provision of services. A Working Party of the Royal College of Physicians. PMID- 7473320 TI - Alcohol liaison in action. The Canterbury experience. AB - In the changing health service, cooperation between disciplines is essential and innovation important to improve the service for people whose health is at risk or damaged. We describe our response to the challenge of liaison between different disciplines when faced with a multifaceted problem that needs the contribution of medical and psychiatric teams and community workers. Although this method of work has been applied to alcohol misuse, it could also be applied to other clinical problems. Liaison can be successful if these is an identified need and the different groups share a desire to work together. To stimulate the liaison and maintain momentum there need to be driving forces, be they clinical, practical or economic, also a shared philosophy and language, a conceptual base in the care of that group of patients, mutual respect and agreed working arrangements. Psychiatry and medicine operating together can ensure that these patients are treated with fairness and respect. They can tap into resources not easily available to either discipline separately, and so create original and exciting services. They can discover other agencies and individuals in their communities who will cooperate with them in obtaining funds and in pressurising statutory bodies to develop much needed services. PMID- 7473321 TI - Copper: not too little, not too much, but just right. Based on the triennial Pewterers Lecture delivered at the National Hospital for Neurology, London, on 23 March 1995. AB - Copper is an essential trace element with a well established mechanism for maintaining balance of the metal in the body, which is controlled by at least two genes. Disruption of one gene on the X chromosome determines a defect in the process of copper absorption, with consequent deficiency of available copper at the cellular level. This results in abnormalities of collagen formation and brain maturation, leading to early death. As yet there is no effective treatment. Disruption of the other copper controlling gene, located on chromosome 13, is associated with accumulation of excess copper in the body, first in the liver leading to cirrhosis, and then in the brain giving rise to destruction of the centre of motor control and, frequently, changes in personality. Copper excess can also cause renal damage, osteoarticular changes and joint pains. If the renal lesion leads to calcium loss in the urine and other tubular defects, there may be osteoporosis or, occasionally, osteomalacia with pathological fractures. In this disease, the abnormal stores of copper can be mobilised with chelating agents or depleted by the administration of zinc salts or thiomolybdate. This usually, but not invariably, leads to improvement or even complete reversal of symptoms. Brain lesions, as demonstrated by computed tomography or magnetic resonance imaging, may also resolve. The mechanism of this phenomenon is obscure but suggests that the long held doctrine that there is no recovery in the central nervous system may have to be reviewed. PMID- 7473322 TI - How do we manage the first seizure in adults? AB - This paper reports an audit of the management of a first generalised seizure in a small district general hospital and describes the management policies of 95 general physicians and 82 neurologists. The audit studied 56 patients who presented in 1990-93 after witnessed generalised convulsions without focal neurological signs or previous history of seizures. Their ages ranged from 16 to 89 (mean 38) and 47 of them were men. In 31% alcohol was incriminated in the seizure. Blood tests were done in most patients but gave little useful information. Skull and chest radiographs were taken in fewer patients but were unhelpful. An electroencephalogram was done in 77% but failed to influence management in any, and only 2 of the 50 computed tomography scans performed led to a change in management. Only 21% received the correct advice about driving. A questionnaire sent to 130 general physicians and 109 neurologists sought their policy on the management of patients after a first generalised seizure. Completed forms were received from 95 physicians and 82 neurologists (response rate 74%). Half of physicians and neurologists perform a computed tomogram on all patients while one-third scan patients only above a certain age. Neurologists perform significantly more electroencephalograms (86% vs 65%) while many more physicians order a chest radiograph (73% vs 16%). About 80% of physicians compared with about 30% of neurologists ask for routine blood tests of haematology and biochemistry. Only 4% of physicians and 1% of neurologists routinely start anticonvulsant treatment. PMID- 7473323 TI - What factors influence patient preferences regarding cardiopulmonary resuscitation? AB - The aims of this study were to investigate the impact of medical and non-medical factors on the cardiopulmonary resuscitation (CPR) preferences of patients, to determine which of them are the most important to patients when considering CPR, and to compare the views of older (> or = 70 years) and younger (< 70 years) patients. We interviewed 180 patients, 86 of whom were aged 70 years or older. 'I do not want to be a burden on my family' was the most important factor for older patients, and they were more ready to leave the decision to the doctor than were the younger patients. 'I want to retain my capacity to think clearly' was most important to younger patients. In general, younger patients gave higher ratings in favour of CPR than older patients. Increased age, drug abuse, dementia, pain, poor functional status and a low likelihood of success were associated with a lower preference rating for CPR in both age groups. PMID- 7473324 TI - Death from phaeochromocytoma: lessons from a post-mortem survey. AB - Despite advances in biochemical assessment and imaging, phaeochromocytoma remains a difficult diagnosis. Using the names of patients whose death certificate listed phaeochromocytoma as a cause of death, a retrospective survey of 62 deaths from phaeochromocytoma (48 benign, 14 malignant) was carried out. All deaths occurred between 1981 and 1989, so the pitfalls uncovered reflect recent practice. A substantial proportion presented with abdominal pain and vomiting, dyspnoea, left ventricular failure or hypotension rather than the classical symptoms. These presentations were more common in this autopsy series than in prospective series of consecutive patients. Diagnosis in the presence of classical symptoms was often delayed but, once it was made, elective excision was relatively safe. A personal or family history of symptoms suggesting inherited diseases associated with phaeochromocytoma was not always given due weight. Biochemical tests, particularly 24 hour urinary vanillyl mandelic acid, often gave contradictory results; the limits of their predictive power should be better appreciated. Anaesthesia and surgery in the presence of undiagnosed phaeochromocytoma was the cause of death in 16 of 62 cases. Recommendations to improve the accuracy of diagnosis are made. PMID- 7473325 TI - Content of a discharge summary from a medical ward: views of general practitioners and hospital doctors. AB - The objective of this study was to seek the views of general practitioners (GPs), hospital physicians and junior hospital doctors about the relative value of different items of clinical information in discharge summaries from medical wards, and so form a minimum and recommended data set for the purposes of clinical audit. GPs were selected randomly from five family health services authorities in England, and hospital consultants and junior hospital doctors were randomly selected from all 14 former health regions. Postal questionnaires were then sent to a sample of 400 GPs, 400 hospital consultants and 400 junior hospital doctors. The results have been tabulated. 'Details of drugs at discharge' (including frequency, dosage and proposed length of treatment), 'significant results of investigations, both positive and negative', 'suggested or made arrangements for follow up', and 'information given to patient about diagnosis' were ranked particularly high by all three groups of respondents. PMID- 7473326 TI - A study of appropriateness of acute geriatric admissions and an assessment of the Appropriateness Evaluation Protocol. AB - We studied the appropriateness of admissions to our acute geriatric unit to determine whether they were clinically indicated and to test the Appropriateness Evaluation Protocol (AEP), an instrument previously validated for general hospital admissions. An admission is appropriate according to the AEP if at least 1 of its 16 criteria is met. At the next ward round, a consultant would judge, given the clinical details available on admission, whether the admission was appropriate, regardless of the AEP. All emergency admissions during a three week period were eligible. Of 186 admissions, 146 (78.5%) were available for analysis. The rates of inappropriateness according to consultants and AEP were 13% and 11% respectively. The AEP had a sensitivity of 97%, specificity of 63%, positive and negative predictive values of 95% and 75% respectively. The overall agreement between consultants and AEP was 92% (kappa = 0.62). There are no data on the optimal level of appropriateness but our rate of 87% (89% using the AEP) appears favourable. This study also shows the AEP to be a suitable tool to monitor the appropriateness of acute geriatric admissions. PMID- 7473327 TI - Why do research ethics committees disagree with each other? AB - Research ethics committees have to ensure that research projects are asking sensible questions and are designed so that the questions will be answered, that the research subjects who are involved will not come to unnecessary harm, and that the autonomy of the research subjects will be respected. Where research proposals cannot fulfill all these criteria, research ethics committees have to perform a delicate balancing act amongst competing moral claims. These arise when, for example, the research is non-therapeutic or the research subjects are incompetent to give consent. Given that the balance of conclusions is so sensitive, it is hardly surprising that different committees sometimes disagree with each other. PMID- 7473328 TI - Setting up interdepartmental peer review. The British Thoracic Society's scheme. AB - In 1992 the British Thoracic Society arranged a series of voluntary interdepartmental peer reviews. Their prime objective was to help to improve services by enabling clinicians to share experiences and ideas on the organisational aspects of the service and training provided. This paper outlines the planning and implementation of the scheme and some of the key results, and discusses the benefits and difficulties of such 'peer reviews'. Much of the benefit comes from the free exchange of ideas at the time of the reviews and 82% of the participants felt that they had picked up new ideas during the review. In their reports, reviewers outlined the strengths and weaknesses of the units and put forward a list of their key recommendations for change. Importantly, in half of the reviews the reviewers identified factors requiring change that had not been anticipated by the reviewed units. In most cases, the reports were subsequently shown to the managers concerned. One year after the review, 53% of 144 major recommendations had been achieved or were imminent. This form of voluntary peer review benefits both the reviewers and the reviewed and could well be adopted by other specialties. PMID- 7473329 TI - Achieving an ethical health service: the need for information. HM Queen Elizabeth the Queen Mother Lecture. PMID- 7473330 TI - Opportunistic infections in HIV seropositive individuals. PMID- 7473332 TI - The 1995 Advanced Medicine Conference. PMID- 7473331 TI - Non-Hodgkin's lymphoma. PMID- 7473333 TI - Clinical computing: friend or foe? PMID- 7473334 TI - Partners in care. PMID- 7473335 TI - Clearing the killing fields. PMID- 7473336 TI - Moses Maimonides, prince of physicians. PMID- 7473337 TI - Management of heart disease in the elderly in the Plymouth health district. PMID- 7473338 TI - The psychological care of medical patients. PMID- 7473339 TI - Mega-trials. PMID- 7473340 TI - Public health implications of ageing. PMID- 7473341 TI - Equity or equality? PMID- 7473343 TI - Effects of prenatal exposure to low-dose beta radiation from tritiated water on the neurobehavior of mice. AB - Pregnant adult C57BL/6J mice, randomly assigned to 1 of 4 groups, 3 of them were irradiated with beta-rays from tritiated water (HTO) by a single intraperitoneal injection on the 12.5th day of gestation. Their offspring received cumulative doses of 0, 5, 10 or 30 cGy in utero. Male pups were trained and examined using a set of behavioral tests that included avoidance acquisition and avoidance maintenance, open field test, hole-board dipping, a water maze, and a food labyrinth. Results were found for most parameters in the 10 and 30 cGy groups that differed significantly from results for the controls, indicating that the behavioral teratogenic effect of prenatal exposure to chronic beta-ray radiation from HTO may be greater than the same dose of acute X- or gamma-irradiation and that 10 cGy may be the lowest detectable dose level at which behavioral changes is detactable under the conditions used in this experiment. PMID- 7473342 TI - More general physicians or specialists? PMID- 7473344 TI - Immediate-early, transient induction of the interleukin-1 beta gene in mouse spleen macrophages by ionizing radiation. AB - In murine spleen cells, x ray irradiation induces the expression of the IL-1 beta gene at multiple phases of the peak time. We analyzed the immediate-early phase of IL-1 beta mRNA accumulation. To determine the lineage of cells that showed the immediate response to irradiation, normal spleen cells were analyzed by Northern blotting and in situ hybridization after separation by magnetic antibodies against specific cell-surface antigens. Although most of the spleen macrophages continuously expressed a low level of IL-1 beta mRNA, a portion of the macrophage population transiently accumulated large amounts of IL-1 beta message immediately after irradiation. A macrophage-like leukemia cell line that resembles these inducible macrophages was identified. A similar immediate-early and transient increase in the IL-1 beta mRNA level occurred when cultured spleen cells were irradiated with a low dose (3 Gy) of x rays. In contrast, the x ray-inducible expression of the IL-1 beta gene was immediate and continuous, not transient, in spleen cells from whole-body irradiated mice. Results of the run-on transcription assay and the determination of the decrease in the message using cultured spleen and macrophage-like leukemia cells indicated that x ray irradiation appears to activate the transcription of the IL-1 beta gene and partially stabilize the message. The results show that the x ray-induced immediate-early accumulation of IL-1 beta mRNA is regulated at both the transcriptional and post-transcriptional levels in an as yet unidentified population of spleen macrophages. PMID- 7473345 TI - Expression of IL-1 beta mRNA in mice after whole body X-irradiation. AB - IL-1 beta is a stimulator of hematopoietic and inflammatory systems, and also acts as a radioprotector. After whole-body exposure to sublethal doses of ionizing radiation, the IL-1 beta mRNA level in spleen cells increases for a short time prior to regeneration of the spleen. We analyzed spleen cells of C3H/He mice after whole-body irradiation with 3 Gy x-rays to determine the cause of this short-term increase in the transcription level. An increase in the level of the message in spleen cells, found by Northern blot hybridization, reached its peak 5 to 7 days after irradiation. There was a low correlation between the curves of the mRNA level and the ratio of monocyte/macrophage lineage cells; a typical source of the message. Spleen macrophages that produce a large amount of the message were found 7 days after irradiation in an in situ hybridization experiment in which heterogeneous spleen cell populations were used. In contrast, spleen cells had no detectable levels of macrophages rich in IL-1 beta mRNA before and 17 days after irradiation. Additionally, the population of message rich cells was 9.4% of the total number of monocytes/macrophages in the spleen. These results suggest that the short-term increase in IL-1 beta mRNA is a result of the heterogeneous differentiation of a subpopulation of spleen macrophages before regeneration of the spleen. PMID- 7473346 TI - Induction of leg-shaking, knock-down and killing responses by gamma-ray irradiation in Shaker mutants of Drosophila melanogaster. AB - Induction of membrane-associated responses, a leg-shaking, a knock-down and a killing, by gamma-ray irradiation was investigated in Shaker (Sh) mutants of Drosophila melanogaster in which the gene cords for the A-current K+ channel. Sh mutants were more sensitive in the knock-down response after gamma-ray irradiation than wild types. There were a great amount of sex difference in the knock-down response, males being more sensitive than females, but not in the killing response. The sex difference was larger than gene dosage effect on X chromosome in females. Genetical analysis revealed that the sensitivity of the knock-down response is an incompletely dominant character without maternal effects. The leg-shaking response, which had previously been reported to be induced by ether treatment, was demonstrated in the head-removed flies of Sh mutants. It was found to be the most sensitive among the responses tested, and may involve changes in K+ channel. The knock-down response may be related to expansion of the leg-shaking response. The killing response should have causes different from the leg-shaking and the knock-down responses judging from the lack of correlation with them. PMID- 7473347 TI - Transfer factors of some selected radionuclides (radioactive Cs, Sr, Mn, Co and Zn) from soil to leaf vegetables. AB - Transfer factors of radionuclides from soil to leaf vegetables (cabbage, Chinese cabbage, komatsuna, spinach and lettuce) have been studied by radiotracer experiments using Andosol as a representative of Japanese soils. The transfer factors of radioactive Cs, Sr, Mn, Co and Zn for edible parts of vegetables (average of five vegetables) were 0.11, 0.24, 0.61, 0.05 and 0.52, respectively. These values should be used in safety assessment for Japanese agricultural environment. The transfer factors of Mn, Co and Zn for spinach were higher than those for the other vegetables. The transfer factors of Cs for different organs of the leaf vegetables were rather homogeneous. The transfer factors of Sr and Mn were higher for older (outer) leaves than younger (inner) ones. In contrast to Sr and Mn, transfer factors of Zn for younger leaves were higher than those for older ones. The distribution ratios of the elements between soil-solution and soil were in the order Sr > Mn > Cs > Co > Zn, whereas the distribution ratios of the elements between plant and soil-solution were in the order Zn > Cs > Mn > Co > Sr. These results indicate that the selectivity for Sr by plants from the soil solution was low and that for Zn was very high. PMID- 7473348 TI - Effect of antihistamines, disodium cromoglycate (DSCG) or methysergide on post irradiation cerebral blood flow and mean systemic arterial blood pressure in primates after 25 Gy, whole-body, gamma irradiation. AB - Exposure to ionizing radiation causes hypotension, cerebral ischemia and release of histamine (HA) and serotonin (5-HT). To investigate the relationship among these responses, rhesus monkeys (Macaca mulatta) received physiological saline (i.v.), disodium cromoglycate (DSCG), antihistamines (AH, mepyramine and cimetidine), or methysergide (METH), then were given 25 Gy whole-body irradiation. Monkeys receiving DSCG, AH or METH had higher post-irradiation mean arterial blood pressure (MBP) than saline-treated controls. Compared to levels in controls, post-irradiation hippocampal blood flow (rCBF) levels were higher in monkeys receiving DSCG, AH or METH. Treatment with the 5-HT2 receptor antagonist methysergide was the most effective in maintaining both rCBF and MBP after irradiation. Results support the hypothesis that the irradiation-induced cerebral ischemia and, to some extent, the hypotension is mediated by serotonin through 5 HT2 receptor sites. PMID- 7473349 TI - Neutron generator at Hiroshima University for use in radiobiology study. AB - A neutron generator (HIRRAC) for use in radiobiology study has been constructed at the Research Institute for Radiation Biology and Medicine, Hiroshima University (RIRBM). Monoenergetic neutrons of which energy is less than 1.3 MeV are generated by the 7Li(p,n)7 Be reaction at proton energies up to 3 MeV. The protons are accelerated by a Schenkel-type-accelerator and are bombared onto the 7Li-target. An apparatus for the irradiation of biological material such as mice, cultured cells and so on, was designed and will be manufactured. Neutron and gamma-ray dose rates were measured by paired (TE-TE and C-CO2) ionization chambers. Contamination of the gamma ray was less than about 6% when using 10 microns-thick 7Li as a target. Maximum dose rates for the tissue equivalent materials was 40 cGy/min at a distance of 10 cm from the target. Energy distributions of the obtained neutrons have been measured by a 3He-gas proportional counter. The monoenergetic neutrons within an energy region from 0.1 to 1.3 MeV produced by thin 7Li or 7LiF targets had a small energy spread of about 50 keV (1 sigma width of gaussian). The energy spread of neutrons was about 10% or less at an incident proton energy of 2.3 MeV. We found that HIRRAC produces small energy spread neutrons and at sufficient dose rates for use in radiobiology studies. PMID- 7473350 TI - [Magnetic resonance imaging]. PMID- 7473352 TI - [Scanners]. PMID- 7473351 TI - [Global view on medical imaging. Apropos of the RSNA 1994]. PMID- 7473353 TI - [Conventional and digital radiology]. PMID- 7473354 TI - [Digital vascular radiology]. PMID- 7473355 TI - [Echography]. PMID- 7473356 TI - [Films and associated devices]. PMID- 7473357 TI - [Novelties in nuclear medicine]. PMID- 7473359 TI - [The point of view of the radiologist]. PMID- 7473358 TI - [Imaging of cerebral function: new trends in PET, MEG and MRI]. PMID- 7473361 TI - [All digital radiology, is it acceptable today?]. PMID- 7473360 TI - [PACS (Picture Archiving and Communication System) networks and teleradiology at the RSNA'94]. PMID- 7473362 TI - [Responsibility and development in medical imaging]. AB - The recent evolution in medical law, permitting a responsibility without objective fault, underlines obligations in radiology. The radiologist is a doctor, specialist, doing an act of radiodiagnosis of which he is responsible, so much in his decision that in his realisation. This concept is important because of the diversity of technical meanings, and the surgical consequences, realised by the radiologist. The medical fault must be examined by the radiologist, by degrees, according to the different responsibilities. Then, we describe a logical realisation of the radiodiagnosis act. At last, we explored the condition of the complain and jurisdictional followings. PMID- 7473363 TI - [Pulmonary embolism: contribution of spiral x-ray computed tomography]. AB - PURPOSE: Spiral computed tomography was compared retrospectively with digital subtraction pulmonary angiography (PA) in 45 patients suspected of having acute or chronic pulmonary embolism. MATERIALS AND METHODS: 45 patients in whom the presence of acute or chronic pulmonary embolism was suspected underwent examination by spiral CT and PA. Diagnosis of pulmonary embolism was based on the direct visualization of intraluminal clots. The study of the agreement between the two methods was based on the Kappa test. In 35 cases, pulmonary emboli were proved. Acute pulmonary emboli were present in 28 cases and chronic in 7 cases. RESULTS: Spiral computed tomography represents an excellent way to detect acute pulmonary embolism. In the chronic form, spiral CT is better than PA to detect intraluminal clots. However, Spiral CT can fail to detect small embole in the peripheral arterial bed. In the 10 patients without pulmonary embolism, the spiral CT proved diagnosis pulmonary oedema (n = 3), lymphangitic carcinoma (n = 4), pleural effusion (n = 3). CONCLUSION: This study suggest that the spiral CT examination is accurate for diagnosis of pulmonary embolism specifically in case of suspected important embolism. The advantages of spiral CT are multiple (non invasive, wide diagnosis spectrum). However, may be a limitation to is use is insufficient distal thrombi detection. This eventuality (5 to 10% in the Pioped study) justify the practice of pulmonary angiography. Spiral CT improvements should reduce this insufficiency in the next future. PMID- 7473364 TI - [Thoracic standard radiology and x-ray computed tomography in mediastino pulmonary tuberculosis in infants]. AB - Primary mediastino pulmonary tuberculosis in infancy is increasing in France. Bacteriological diagnosis is difficult. The aim of the study is to assess the value of chest radiograph and CTscan for diagnosis of tuberculosis. We studied 15 infants, 3 month old to 18 month old, between November 1990 and June 1992. On plain film, lymphadenopathies were present in 13 cases and parenchymal disease in 10 cases, parenchymal nodular densities in 6 cases and pleural effusion in one case, displacement of the trachea in 6 cases. We found CTscan to be useful in delineating the lymphadenopathies and parenchymal changes. In 2 cases lymphadenopathies and in 2 cases alveolar condensation were discovered on CTscan as plain film was normal. In all cases lymphadenopathies were present in CTscan, often in right side and in all cases, lymphadenopathies were hypodense with peripheral enhancement. In conclusion, the association of right paratracheal nodes with displacement of trachea and alveolar consolidation is common in tuberculosis in infants. These findings when seen on chest radiographs must lead to perform a CTscan and suggest the diagnosis. PMID- 7473365 TI - [Unilateral inguinal hernia in infants: costs, risks and benefits of herniography? Methodology]. AB - AIM: To assess the validity and the interest to health of systematic herniography in cases of unilateral inguinal hernia in children under two years of age. METHOD: Decision theory was used to assess the effect of herniography on individual health by estimating gonadic benefit and establishing a benefit-risk ratio, and on collective health by the use of cost-effectiveness analysis. RESULTS: a decision tree and algorithms were developed. DISCUSSION: Decision analysis is used in complex medical decision making, clarifying choices and subdividing a problem into several more manageable sub-problems. The threshold approach pinpoints factors requiring more information. PMID- 7473366 TI - [Unilateral inguinal hernia in infants: costs, risks and benefits of herniography? Results]. AB - AIM: To assess the advantages of systematic herniography in cases of unilateral inguinal hernia in infants. METHOD: Decision analysis presented in the first part of this article was used. A preliminary retrospective study of 348 cases as well as a literature review were utilized to determine the probabilities required in order to create and run the decision aid algorithm. Gonadic benefit was used as the health indicator. RESULTS: For males, gonadic benefit procured by the herniography depends on the risk of strangulation in cases of groin hernia and on the frequency of testicular atrophy after inguinal hernioplasty. The evaluation of these two elements largely determines the choice of medical and surgical practices. For a 0.44% risk of post-operative testicular atrophy and a risk of strangulated hernia estimated at 20%, the cost-effectiveness ratio is FF, 199681 to save a testicle. One testicle is saved every 455 herniographies, with 24 possible post-examination complications. For female, herniography provides no gonadic benefit. DISCUSSION: Decision analysis permits the quantification of results, leading to improved clinical judgement and facilitating the evaluation of medical practice. PMID- 7473367 TI - [Hemoperitoneum caused by spontaneous rupture of hepatocellular carcinoma. Value and prognostic factors of hepatic artery embolization]. AB - To assess efficiency and to determine prognostic factors in emergency embolization for intraperitoneal hemorrhage from hepatocellular carcinoma we retrospectively analyzed 12 consecutive procedures. In order to evaluate the extension of tumor thrombus in the intrahepatic portal system before embolization, a portal perfusion rate was defined as the relative patency of the segmental portal veins demonstrated at arteriography. Serum bilirubin level and portal perfusion rate were correlated with length of survival. Successful hemostasis was achieved in all patients. Mean length of survival was 149 days. Two patients are still alive 545 and 195 days, respectively, after the procedure. A significant correlation (p = 0.005) between portal perfusion rate and length of survival was obtained. There was no correlation between serum bilirubin level and prognosis. Emergency embolization is an effective treatment in patients with intraperitoneal hemorrhage from hepatocellular carcinoma. Prognosis depends of portal perfusion rate. PMID- 7473368 TI - [Mucormycosis: an uncommon case of cellulitis of the face and multiple facial lymphadenopathies]. AB - An unusual case of mucormycosis is presented: a cutaneous form characterized by an extension to the soft tissues of the face with a noticeable involvement of facial lymph nodes. The successive CT studies enable the description of the lesions and their follow-up. PMID- 7473369 TI - [Spinal aneurysmal cyst]. AB - The authors report one case of aneurysmal rachidian cyst who concern a young girl of eleven years old presenting lumbagos with pyramidal irritation on a L2 and L3 lesion. The radiological exploration (CT, MRI) has a double interest: to find again lytical pictures of the rear arch blowing the cortical, liquids levels, logets which remind the macroscopical appearance of the aneurysmal rachidian cyst; to specify the regional extension of preoperative interest. The histological confirmation of the aneurysmal rachidian cyst after a surgical treatment and the conformity of anatomo-radiological descriptions, confirm the predictive value of the semeiology described in case of aneurysmal rachidian cyst. PMID- 7473370 TI - [Dental-radicular cysts invading the maxillary sinus]. AB - The authors report three cases of radicular cyst developing into the maxillary sinus which was partially or completely obscured. Looking for a thin rim of cortical bone should be undertaken in case of opacity of maxillary sinus. This rim correspond to the floor of the sinus, which is lifted up by the cyst. If the attenuation value of the intrasinusal process is near of that of water, the diagnosis of radicular cyst invading the maxillary sinus is very likely and may indicate intraoral surgical approach. PMID- 7473371 TI - [Tracheal carcinoma infiltrating the thyroid gland and mimicking subacute thyroiditis. Contribution of Doppler to differential diagnosis]. AB - In the present case Color Coded Doppler showed a marked increase in vascularization in the right lobe of the thyroid with high systolic velocities (1 m/s). This sign was only doubtful compared with clinical, biological and isotopic data. Because of dramatic laryngeal dyspnea, surgery was conducted which allowed the final diagnosis of thyroid invasion by tracheal epidermoid carcinoma. This demonstrates the particular role of Color Coded Doppler in the management of inflammatory thyroid diseases. PMID- 7473372 TI - [Cutaneous appearance of dorsal spinal cord lesions in MRI]. PMID- 7473373 TI - [Superinfection in chronic ischemic colitis. A radiological case]. PMID- 7473374 TI - [Congenital neck masses. Embryonic origin and diagnosis. Report of the CIREOL]. AB - Various congenital cervical anomalies are found in the neck region including defects of the branchial apparatus (branchial, thymic and parathyroid anomalies) and vascular anomalies. They manifest as cystic masses, sinuses, fistulas and as ectopic glands. This multicentric retrospective imaging study done in 5 different radiological centers (4 adult radiological departments and 1 pediatric radiological department) shows the result in understanding the congenital cervical anomalies and include 63 patients. The age of the patients varied between 24 days-81 years with a mean age of 23 years. This study included 27 patients having congenital branchial pouch anomalies (4 cases of anomalies of obliteration of the 4 th arch), 14 cases of cervical cystic hygromas, 11 thyroglossal tract cyst cases, 1 congenital laryngocele case, 1 case of jugular ectasia, 3 cases of capillary haemangioma. The embryologic basis of these different malformations were reviewed. Their characteristic findings and sites were illustrated together with their typical et atypical appearances. The frequency of occurrence of each branchial anomaly were plotted, the second branchial cleft cyst being by far the most common congenital cystic neck mass (70%). The study revealed the role of different imaging modalities in the diagnosis of various congenital cervical anomalies, especially in some particular complicated cases of congenital neck masses presenting in adult. Imaging study helps the clinician to anticipate any difficulties in unforeseen circumstances that may arise including infection haemorrhage, or parapharyngeal extension. Understanding the various radiologic appearances of these anomalies is greatly aided by familiarity with their embryologic origin. Moreover, considering the anatomic location and radiologic appearance, the precise embryologic origin can be accurately predicted. PMID- 7473375 TI - [Comparative studies of the tolerability of gadodiamide, dimeglumine gadopentetate and meglumine gadoterate in MRI tests of the central nervous system]. AB - The development of new non-ionic magnetic resonance (MR) contrast media as gadodiamide injection increased the choice of paramagnetic contrast agents available in MR of the central nervous system (CNS). The purpose of our paper was to compare at the dose of 0.1 mmol/kg b.w. the safety of gadodiamide (Gd-DTPA BMA) to gadopentetate dimeglumine (Gd-DTPA) and to gadoterate meglumine (Gd-DOTA) in two multicentric double-blind studies. A total of 551 patients were enrolled with 143 patients in the Gd-DTPA group, 132 patients in the Gd-DOTA group and 276 patients in the Gd-DTPA-BMA group. Safety was assessed by recording the adverse events up to 24 hours after the injection. One or more adverse events were recorded in 14% of the Gd-DTPA patients, in 15.1% of the Gd-DOTA patients and in 11.6% of the Gd-DTPA-BMA patients. These reactions were related to the contrast media in 9.1%, 13.6% and 8.7% of the cases respectively. Their intensity was defined as mild in 8.4% of the patients in the Gd-DTPA group, in 13.6% of the patients in the Gd-DOTA group and in 8.3% of the patients in the Gd-DTPA-BMA group. No severe reaction or death were recorded. An injection-site reaction (heat, coldness, pain) has been observed in 43% of the cases although an adverse event other than local reactions (headache, dizziness, nausea) has been noticed in 57% of the cases. No significant statistical difference was observed between the groups. Gadodiamide is a safe and effective contrast agent in MRI of the CNS in comparison with Gd-DTPA and Gd-DOTA currently in routine use. PMID- 7473376 TI - [Electrocorticographic evaluation of the neurologic tolerability of iobitridol (Xenetix), a new non-ionic contrast medium in rabbits]. AB - Electro-corticographic tracings (two longitudinal leads, bipolar assembly) were recorded from curarized rabbits (5/group) receiving selective internal carotid artery injections of either iobitridol, a new non-ionic contrast medium, or iohexol, the high osmolar diatrizoate or hypertonic mannitol (isotonic to the non ionic agents). A further group was submitted to the surgical preparation but was not injected. The solutions were injected at a dose of 2.5 ml during 30 seconds. The animals were anaesthetized (halothane) during the surgical period. The permeability of the blood-brain barrier was assessed by means of the extravasation of Evans'blue. Tracings were visually assessed and a semi quantitative method for blind evaluation of fast/slow rhythms was used. This method was pharmacologically validated by the use of pentobarbital and pentylenetetrazole. Diatrizoate and iohexol induced respectively 3 and 2 paroxystic tracings during or immediately after the injection period. Iobitridol and hypertonic mannitol did not cause such effect. Paroxystic tracings in the iohexol group were not associated with extravasation of Evans'blue in the cerebral parenchyma. Blood brain barrier was disrupted in all rabbits receiving the high osmolar agent diatrizoate. Tracings of the control group were characterized by a progressive increase of fast rhythms, as those of the iobitridol and mannitol groups. On the contrary, iohexol and especially diatrizoate induced an increase in the proportion of slow waves. Taken together, these data suggest that iobitridol shows an excellent tolerability potential for clinical use. PMID- 7473377 TI - [Clinical evaluation of the tolerability of gadodiamide, a new nonionic contrast agent in MRI of the central nervous system]. AB - Gadodiamide injection (Gd-DTPA-BMA) is a new non-ionic paramagnetic contrast agent for which the safety at the dose 0.1 mmol/kg was evaluated during a European multicentre study on a large population of adult patients who had an MR examination of the central nervous system with contrast medium. The safety analysis was performed on 2,102 patients by recording the adverse events observed during injection and up to 24 hours after the injection. Adverse events due or probably due to gadodiamide injection were observed in 102 patients (4.4%) with injection-site associated discomfort (heat, coldness, pain at the injection site) in 37 patients (1.8%) and adverse events other than discomfort (headache, nausea, vomiting) in 35 patients (3.1%). No adverse events of severe intensity or death were reported during the trial. Gadodiamide injection was shown to be safe and well tolerated and represents a non-ionic alternative to the current products in the field of MR imaging of the central nervous system. PMID- 7473378 TI - [Hypersensitivity pneumopathies: x-ray computed tomographic aspects]. AB - Chest radiographs and computed tomographic scans obtained from nineteen patients with hypersensitivity pneumonitis were reviewed retrospectively. Thirteen of them had a subacute form. Six of them had a chronic form. In subacute forms, CT scans were normal in two patients. In the 11 other cases, CT findings were ground glass attenuation (n = 10), nodules (n = 7) and linear opacities (n = 5). Ground glass attenuation was diffuse or patchy. Two patterns are quite suggestive of hypersensitivity pneumonitis: the association of micronodules and ground glass attenuation and the presence of hyperlucent secondary lobules within the ground glass. After cessation of exposure, CT showed a return to normal or improvement. Ground glass opacities, nodules and also linear opacities disappeared. CT scans were always abnormal in chronic forms. CT findings were linear opacities (n = 6), ground glass attenuation (n = 4), micronodules (n = 2) and cysts (n = 3). PMID- 7473379 TI - [Value of calculation of the kappa coefficient in the evaluation of an imaging method]. AB - Radiologists are often confronted to the choice of the most appropriate statistic tool for evaluating diagnostic imaging methods. Using a radiological literature example, the Kappa agreement test is herein described and its applications are determined. Although widely performed to determine the inter-rater agreement, this test is also suited for the confrontation of two or more diagnostic imaging methods applied on the same subjects and providing categorical data. It procures the degree of agreement between the different methods. PMID- 7473380 TI - [Preoperative diagnosis of xanthogranulomatous cholecystitis]. AB - Xanthogranulomatous cholecystitis (XGC) is a benign chronic inflammation of the gallbladder, rarely described in the radiologic literature. Like xanthogranulomatous pyelonephritis, it can clinically and radiologically mimic carcinoma. This unusual entity is characterized morphologically by a broad spectrum of xanthogranulomatous changes seen from a small limited focus within yellow nodule in the gallbladder wall, to diffuse involvement of the entire gallbladder with extension of the fibrosis into surrounding tissues. It is clear that recurrent inflammation and calculi are important for the pathogenesis, which is not well understood. The clinical presentation and radiologic findings of XGC are non specific. Irregular thickening of the gallbladder wall and local extension of the process can mimic carcinoma. Diagnosis of XGC is always established by histological examination, characterized by the infiltration of round cells, lipid laden histocytes and multinucleated giant cells in the muscle layer. We report a case of 76-year-old woman who had an episode of epigastric and right upper quadrant pain, 4 months before admission. Physical examination demonstrated a palpable mass in gallbladder region. Echography and computed tomography showed a large gallbladder, a thickened wall and an infiltration of the adjacent liver. The relatively well defined gallbladder internal border and the absence of biliary tract's dilatation allowed us to suggest the diagnosis of XGC, which was confirmed intraoperatively by frozen section histology. Once the diagnosis was established, cholecystectomy was performed. Occasionally, the inflammatory reaction is so severe that a subtotal cholecystectomy is required. Postoperative recovery was, as usually, uneventful. Although a rare entity, XGC should be considered in the differential diagnosis of complex right upper quadrant masses, as well as neoplastic gallbladder disease. PMID- 7473381 TI - [Circumscribed myositis ossificans: bone changes detected by MRI]. AB - The authors report a case of ossificans myositis, in which MRI showed inflammatory changes of the adjacent bone. T1 weighted fat saturation sequence with gadolinium injection showed enhancement of medullary and cortical bone. This potentially mistaking pattern must be known, to avoid misdiagnosing with malignant osseous tumor, specially before achievement of the characteristic pattern of zonal maturation and its calcified rim. PMID- 7473382 TI - [Spinal cord toxoplasmosis and AIDS]. AB - Toxoplasmosis is the most common brain parasitic infection in acquired immunodeficiency syndrome (AIDS). Spinal cord localisations are still rare (2 cases with cerebral involvement, 2 cases without). A case of both spinal cord and cerebral involvement is reported. MR imaging was performed because of sensory level (L1). A focal conus medullaris enlargement was seen, iso intense on T1 weighted images. This lesion was hyperintense on T2 weighted sequence, and was homogeneously enhanced after Gadolinium on T1 weighted images. A medullary oedema was noted. A toxoplasmosis treatment was initiated, without corticotherapy. MR imaging performed one month later (D30), while important clinical improvements were seen, pointed out normal thickness of conus medullaris, without enhancement after Gadolinium. Disease lesions in AIDS with focal spinal cord processes are reviewed, and diagnostic work-up is discussed. Spinal cord single lesion, associated or not with brain involvements should be treated as a toxoplasmic infection, with MR imaging follow up. This work up should avoid medullary biopsy, still required in case of treatment failure. Cerebral involvements, with multiples lesions, can mask medullary localisation. PMID- 7473383 TI - [Economic stakes and prescription of iodinated contrast media]. PMID- 7473384 TI - [Disappearance of a 35 mm film in coronarography: chronicle of an announced demise]. PMID- 7473385 TI - [Radiologists: moods and questions]. PMID- 7473386 TI - [Contribution of the cervical scanner with injection in the diagnosis of vertebral artery dissection]. AB - Vertebral artery dissection is the first diagnosis that must be suspected by a 40 years old patient with posterior fossa ischemic signs. CT data from six cases of spontaneous or post-traumatic vertebral artery dissections are reviewed. Cervical CT is performed after contrast medium injection from C7 to C0 with 1.5 mm thick slices and a 2 mm gap or with a spiral mod including millimetric reconstruction. The enhancement of the vascular wall, the hypodense hematoma surrounding a stenotic and eccentric lumen, and the enlargement of the artery are typical for a vertebral artery dissection. CT findings were correlated with angiography that is still considered to be the gold standard for this diagnosis. Among noninvasive technics including MRI, duplex scanning and CT, the latter appears very accurate to diagnose vertebral artery dissection. PMID- 7473387 TI - [Septic thrombosis of the cavernous sinus complicating malignant staphylococcal infection of the face]. AB - Septic thrombosis of the cavernous sinuses most commonly follows staphylococcal infections of the middle third of the face. Orbital symptoms are constant. We report six cases diagnosed only by CT. At present, MR imaging is the diagnostic procedure of choice. Its findings are thought to reflect the septic nature of the thrombosis. PMID- 7473388 TI - [Role of MRI in spinal and spinal cord emergencies]. AB - Out of 137 cases concerning the spine and spine cord, the authors demonstrate that MRI is nowadays fully integrated in urgency investigations. In all cord compression suspicions and in spine trauma investigations, it's place is found. Coil and software improvements allow to put CT and MR scanning delays on a line. This study underlines the necessity of being equipped with non magnetic life support systems allowing management of very fragile patients. In traumas, MR provides the lesion's authentification, it's type, it's unique localisation or not, and therefore an early functional prognosis. The discovery of spine cord involving lesions in neoplastic conditions allows to follow a therapeutic strateging for maintaining the motor function. In the end, the authors discuss about practical utility of MR in scuba diving decompression accidents. No lesion of the spine cord has been demonstrated in 25 patients, probably because of technical insufficiency. PMID- 7473389 TI - [The double band sign: pleural tuberculosis in x-ray computed tomography]. AB - We described on computed tomography, two cases of pleural tuberculosis with parietal pleural enhancement and thickening associated with another external high density line. We called this parietal pleural and extra pleural change, the "double band sign". After study of the regional anatomy, we think that this external line is the endothoracic fascia. PMID- 7473391 TI - [Post-radiation necrosis of the cricoid cartilage: an uncommon case]. AB - Radiation therapy is considered as the treatment of choice for early stage laryngeal cancer. However, a few patients, after several years, develop severe life threatening complications, like edema or chondronecrosis. The clinical examination or endoscopy follow up may be difficult. CT and MR imaging can show the exact diagnosis depicting exquisitely the anatomy of the larynx. But at the beginning, the abnormalities are radiographically subtle and their diagnosis may be difficult because of artefacts due to patient dyspnea and mucous secretions. The main suggestive imaging features for suspecting a chondronecrosis seem to be the lost of normal signal hyperintensity on T1 MR imaging or a focal lack of cortical bone on CT scans, just adjacent to a focal swelling of the pharyngo larynx mucosa. The barium pharyngography remains the best imaging diagnosis procedure for depicting associated abnormal communication between the digestive tract and the adjacent spaces or the cartilages of the larynx. Therefore, when the diagnosis is delayed, only salvage total laryngectomy can stop the infectious process. We report one case of a cricoid cartilage chondronecrosis eleven years after radiation therapy for laryngeal cancer. PMID- 7473390 TI - [Hepatosplenic silicosis]. AB - A case of hepatosplenic silicosis complicating a pulmonary silicosis is reported. Numerous calcified splenic nodules seen with conventional radiology, ultrasound or computed tomography, hepatic microcalcifications and "egg shell" abdominal lymph nodes best seen with computed tomography represent the radiologic signs. However, hepatosplenic silicosis can only be confirmed by pathological examination of liver biopsies showing birefringent particles within hyalinized nodules. PMID- 7473392 TI - [Diagnosis of extrapontine myelinolysis preceding central pontine myelinolysis]. AB - An MR study performed on a patient who presented a tetraplegia after liver transplantation, revealed bilateral hypersignal of internal globus pallidus on T2 weighted images. Diagnosis of extrapontine myelinolysis was proposed based on clinical presentation and lesions topography, although absence of natremic disorders. Confirmation of diagnosis was obtained on an MR study performed a month later, which demonstrated a pontine extension. Extrapontine myelinolysis can precede pontine involvement. Diagnosis of myelinolysis should be evoked although MR normal appearance of the pons. PMID- 7473393 TI - [Imaging of osteomedullary infarctions and their complications]. AB - Radiographic, CT, bone scan and MR patterns of the infarcts of the metaphyses and diaphyses of tubular bones and their main complications are presented. Bone marrow infarcts are located in the medullary cavity of long bones. They most often involve the lower limbs. They are commonly associated with necrosis of the epiphyses and have common origin. In contrast to necrosis of the epiphyses, bone infarcts may present specific complications. They are most often detected incidentally, because of their clinical latency. Radiographic and CT changes appear late, showing localized areas of increased density with irregular rims, or sharply demarcated shells of calcification. Earlier changes are demonstrated by bone scan and MRI. MR pattern is characteristic, showing an area of hyposignal on T1- and T2-weighted sequences, with irregular rims, and sometimes small areas of fat signal. Their complications include cystic formation and malignant degeneration (sarcoma and particularly malignant fibrous histocytoma). An unusual complication, infection involving bone infarcts is also presented. PMID- 7473394 TI - [Endoluminal 3D reconstruction of the tracheo-bronchial tree (virtual bronchoscopy)]. PMID- 7473395 TI - [All you want to know about the Clinton reform without ever having the courage to ask. Presentation of the principles and necessary glossary for understanding the jargon]. PMID- 7473396 TI - [Imaging of osteosclerotic metastases]. AB - Carcinomas of the prostate and breast are the most common sources of osteosclerotic metastases. The osteoblastic response is related to stromal bone formation and reactive bone formation. Purely osteosclerotic or mixed osteolytic osteosclerotic lesions are encountered. Frequent differential diagnoses are enostoses and Paget's disease. The classical MRI pattern is a signal of low intensity on T1 and T2- weighted sequences, but some osteoblastic metastases have an inhomogenous signal. Evaluation of the response to treatment is difficult and requires confrontation with clinical and biological data. PMID- 7473397 TI - [Anatomy of the circle of Willis with 3D time of flight magnetic resonance angiography and analysis of partitions]. AB - The purpose of this study was to analyse with 3D TOF MR angiography the anatomical variants of the circle of Willis (CW) in 109 consecutive patients. In all cases, the quality of exams was sufficient to analyse the anatomy of CW. Nevertheless, MRA over estimated hypoplastic segments because incomplete CW were more frequently demonstrated with MRA than in anatomic literature. This lack of MRA sensitivity was related to artifacts as MIP artifacts and blood flow artifacts. MRA sensitivity was improved by analysis of partitions with MIP reconstructions. PMID- 7473398 TI - [MRI of the semicircular canals in cranio-facial asymmetries and craniomandibular syndrome]. AB - The cranio-mandibular syndrome (CMS) is often secondary to an cranio-facial asymmetry (CFA); it associate a painful masticatory dysfunction syndrome with functional and aching clinical signs in relation with a functional torticollis required for oculo-labyrinthal equilibration maintenance. Our anatomical and functional MRI protocol uses tridimensional study of semi-circular canals (SCC) up relation to the medial cerebral axis (MCA), and permit to measure precisely these CFA and propose, with a multispecialist approach, an adapted treatment of the SCM. Application of MRI to SCC asymmetry looks to its future use for spatial intracranial referencial of cerebral structures. PMID- 7473399 TI - [Value of the "Dentascanner"al in dento-maxillary diseases (infectious and cystic)]. AB - MATERIAL AND METHODS: From July 1993 to April 1994, forty dentascans were performed at Bicetre Hospital, on an Elscint Elite Plus scanner for: chronic sinusitis (18 cases); cystic pathologies (8 cases); pre-operative assessment for dental implantation (14 cases). RESULTS: Dentascan images permitted a precise analysis of dentomaxillary anomalies. In all cases they demonstrated the anatomical relationships of the lesions, permitting a determination of potential complications. In chronic sinusitis dental pathology was demonstrated including peri-apical granulomas or cysts breaks or elevations of the floor of the maxillary sinus, malplaced dental filling material in the maxillary antrum and fistula tracks. Among the 14 pre-operative assessments, four cases of infectious lesions were demonstrated, thereby counter-indicating dental implantation. CONCLUSION: The dentascan appears to be very useful for dentomaxillary pathology by facilitating diagnosis and the formulation of treatment and operative strategies. PMID- 7473400 TI - [Spontaneous disappearance of herniation pit on the femoral neck]. AB - The authors present a case of a 39 years old patient who had a herniation pit of the left femoral neck, 5 years ago. This lesion disappeared spontaneously. To our knowledge, no report of healing of a herniation pit has been previously described. PMID- 7473401 TI - [Imaging of lipoma of the pericardium]. AB - Pericardial lipoma is a benign mediastinal tumours. Its location into the pericardial space is well demonstrated by MRI or CT if pericardium is visible; otherwise, diagnosis is evoked on the displacement or compression of cardiac cavities. PMID- 7473402 TI - [Cat-scratch disease presenting as a pseudotumor. Imaging by x-ray computed tomography and MRI]. AB - Cat-scratch disease is usually a self-limiting illness but it can initially present as a tumor. A case of a lesion of the arm is described, with CT and MR findings. The radiologic appearance is not characteristic but the diagnosis can be suggested in a predisposing clinical setting. PMID- 7473403 TI - [Normal and pathological MRI aspects of the posterolateral corner of the knee]. AB - The purpose of the study is to compare normal PLC anatomy and its MRI appearance, with the various lesions observed in MRI, from the simple popliteus tendinous contusion to the complete PLC rupture. For this specific work on PLC lesions, we selected 61 examinations among the traumatic knees explored during the last 3 years. Surgical correlation is obtained for the 61 patients. MRI examinations are performed on a 0.5 T. unit with gradient echo T1, T1 and T1 GD-DOTA IV. Normal PLC anatomy is compared to the dissection of 4 anatomic subjects. Normal MRI slices are evaluated with this reference analysis. The principle anatomical structures of the PLC include the lateral collateral ligament, the popliteus tendon, the arcuate ligament, the fabello fibular ligament, the posterolateral condylar capsule, and the posterior horn of the lateral meniscus. Surgical findings confirm PLC lesion for 58 patients with 3 false positive. Diagnosis of these lesions is important because chronical posterolateral laxity is secondary to the destabilisation of lateral condyle. Unrecognised and untreated posterolateral instability may result in failure of ACL reconstruction. When clinical tests are doubtful or complex, or the examination very painful, MRI evaluates completely the traumatic knee and particularly the PLC. PMID- 7473404 TI - [The temporomandibular joint meniscus in MRI]. AB - The continuing improvement of the MRI's equipment and software led us to a continuous adaptation of the exam technique. We use today a bilateral coil technique, 3 inches in diameter. Four slices are performed for each articulation (4 mm thick) in the true sagittal plane. A fast spin echo, balanced in proton density, for one minute, allows the study of 15 steps inside a full open/close cycle. Oblique coronal cuts are also performed parallel to the long axis of the condyle in a T1-weighted spin echo sequence in close and half-open positions. The analysis of 350 subjects allows to detail more precisely the meniscal displacement. Beside the classic anterior luxation, the meniscus slips medially, laterally and may rotate. These different situations may be isolated or combined. The dynamic study is specifically useful in the displacement with reduction exams. It enables permits to point out the timing of the disk disturbance: this information may be of great help in therapeutic decisions. PMID- 7473405 TI - Peripheral excitatory effects of two enkephalinase inhibitors, acetorphan and thiorphan, and an enkephalin analogue, [D-Ala2-Met5]-enkephalinamide, on uterine motility in periparturient rats in vivo and in vitro. AB - The effects of two enkephalinase inhibitors, acetorphan and thiorphan, and the enkephalin analogue [D-Ala2-Met5]-enkephalinamide (DAMEA), on spontaneous uterine contractions were studied at day 21 of pregnancy in rats following treatment in vivo or in vitro. Acetorphan (10 mg kg-1) and thiorphan (1 mg kg-1), immediately after their i.v. administration, increased the duration of spontaneous contractions 3.4- and 4.6-fold, respectively, but did not modify the maximum amplitude. Similarly, thiorphan (40 mumol l-1) increased the duration of contractions when administered in vitro. Thiorphan was ineffective during the first 30 min when given into the cerebral ventricles (50 micrograms per rat). These results suggest that the enkephalinase inhibitors are acting via a peripheral opioid pathway; and this conclusion is supported by the observation that thiorphan potentiated the stimulatory effect of a submaximal dose of DAMEA administered in vitro. The excitatory effects of DAMEA and the enkephalinase inhibitors were blocked by naloxone. This antagonistic effect of naloxone on uterine motility in the periparturient rat uterus, induced by either acetorphan and thiorphan or DAMEA, seems to be regulated by peripheral opiate receptors. Naloxone (10 mg kg-1 s.c.) increased both the amplitude and duration of uterine motility in vivo; however, naloxone (26 mumol l-1 and 52 mumol l-1) produced a paradoxical dose-dependent biphasic effect in vitro. PMID- 7473406 TI - Annual changes in serum concentrations of prolactin in captive male black bears (Ursus americanus). AB - Prolactin may be involved in the regulation of reproduction in black bears (Ursus americanus) as it is a mediator of photoperiodic changes in a number of species. The objectives of this study were to validate a radioimmunoassay to measure prolactin in bear serum and to describe seasonal changes in serum prolactin concentrations in captive male bears. Serum samples were obtained nine times during a year from three captive male black bears that were denning between November and March and active during the other months. The heterologous prolactin radioimmunoassay, using pig 125I-labelled prolactin and goat anti-pig prolactin as a primary antibody, was validated. Injection of thyrotrophin-releasing hormone into the three male bears in June resulted in a rapid increase in serum concentrations of prolactin (t = 0, 11.4-14.8 ng ml-1; t = 15-30 min, 18.4-28.7 ng ml-1). The sensitivity of the assay was 0.08 ng per tube. Intra- and interassay coefficients of variation were 5.5% (n = 6) and 5.7% (n = 6), respectively. Serum concentrations of prolactin changed seasonally, with the lowest concentrations in December (mean +/- SD = 1.1 +/- 0.1 ng ml-1); this was followed by a gradual increase between January (2.6 +/- 0.6 ng ml-1) and April (6.4 +/- 1.2 ng ml-1) and the highest concentrations in May (17.6 +/- 4.7 ng ml 1), preceding peak testosterone concentrations in June. The observation that prolactin secretion increased with increasing daylength suggests that photoperiod may be an external regulator. The presence of high concentrations of prolactin before peak testosterone concentrations suggests that prolactin may play a role in regulating seasonal changes in the testes. PMID- 7473407 TI - Total number of immunocompetent cells in the normal rat epididymis and after vasectomy. AB - The aim of this study was to make unbiased quantitative estimates of the number of T8 cytotoxic cells and T4 helper cells and macrophages in the head and tail of rat epididymides, using newly developed design-based stereological methods in fertile control animals, and to compare these with results from vasectomized animals. The location of these cells within the intra- and interepithelial compartments of the epididymis was also studied. In control animals, both the T4 helper cells and macrophages and T8 cytotoxic cells were found in significantly higher numbers (P < 0.01) in the head interepithelial location than in the intraepithelial site. In addition, there were significantly more (P < 0.01) T8 cytotoxic cells in the tail interepithelial region. Within the head region of vasectomized rats, there was a significant increase in T8 cytotoxic cells in the intraepithelial location (P < 0.01), and a decrease in T4 helper cells and macrophages in the interepithelial region (P < 0.05). Serum samples from vasectomized animals, when compared with those from control animals, showed a significant increase in cytotoxic antisperm antibody. This suggests that, in response to the increased spermatozoal antigenic absorption that takes place in the epididymis after vasectomy, the immune system redistributes its T cell subsets to modify the immune tolerance that may occur. PMID- 7473408 TI - Response of prolactin to different photoperiods after surgical disconnection of the hypothalamus and pituitary in sheep fetuses. AB - The aim of this study was to determine the impact of surgical disconnection of the fetal hypothalamus and pituitary on the fetal prolactin response to different photoperiods. Disconnection of the hypothalamus (HPD) or a sham operation was carried out at around day 110 of gestation (term = 145 +/- 3 days). Before surgery, pregnant ewes were maintained under a photoperiod of 12 h light:12 h dark. After surgery, ewes carrying HPD fetuses (n = 10) or intact fetuses (n = 13) were exposed to either a long day (16 h light:8 h dark) or a short day (8 h light:16 h dark) regimen until day 143 of gestation. Thyrotrophin-releasing hormone (50 micrograms) was administered intrafetally at days 130-135 of gestation and chlorpromazine (a dopaminergic antagonist) was administered to all sheep fetuses at days 141-142 of gestation. Mean fetal prolactin concentrations were significantly higher in the long day group (HPD: 37.3 +/- 11.3 ng ml-1; intact: 71.0 +/- 16.2 ng ml-1) than in the short day group (HPD: 9.0 +/- 4.8 ng ml-1; intact: 34.2 +/- 16.0 ng ml-1). In the intact group, fetal prolactin concentrations increased significantly between day 6 and day 30 of exposure to either photoperiod. However, in the HPD group, fetal prolactin increased with increasing exposure to the long day photoperiod and decreased with increasing exposure to the short day photoperiod.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7473409 TI - Increased numbers of Sertoli and germ cells in adult rat testes induced by synergistic action of transient neonatal hypothyroidism and neonatal hemicastration. AB - The combined effects of transient neonatal hypothyroidism and neonatal hemicastration were investigated to see whether they were additive. Hypothyroidism was induced in litters of ten male rats for 25 days from the day of birth by administration of 0.1% (w/v) 6-propyl-2-thiouracil in the mother's drinking water; hemicastration was performed on the day of birth. Controls included both normal and sham-operated animals. Numbers of Sertoli cells and round spermatids were quantified at age 135 days using stereological methods. Sham-operation had no effect on testis mass, or numbers of Sertoli or germ cells. Transient neonatal hypothyroidism resulted in an increase in testicular mass of 27% (P < 0.05), whereas neonatal hemicastration resulted in a 33% (P < 0.05) increase over control; the combination of the two procedures resulted in a 62% (P < 0.05) increase. There were corresponding significant increases in the number of Sertoli cells: 82% with hypothyroidism, 18% with hemicastration and 123% with the combination of the two procedures. Numbers of round spermatids showed similar increases: 59% with hypothyroidism, 45% with hemicastration and 95% with the combination of the two procedures. It is concluded that the effects of the combination of transient neonatal hypothyroidism and hemicastration are additive with respect to testicular mass, and numbers of Sertoli and germ cells. PMID- 7473410 TI - The effect of duration of exposure to short days on the gonadal response to long days in male starlings (Sturnus vulgaris). AB - Photorefractory male starlings (Sturnus vulgaris) were transferred from a photoperiodic schedule of 18 h light:6 h dark to 8 h light:16 h dark for 2, 4, 6 or 10 weeks, and then photostimulated by exposure to 18 h light:6 h dark. Testicular dimensions were measured at regular intervals by laparotomy. There was no change in testicular volume in birds photostimulated after 2 weeks under a photoperiod of 8 h light:16 h dark, but a cycle of testicular growth followed by regression occurred in the other groups. Testicular volume was increased significantly by 20 days under 18 h light:6 h dark in all three groups; regression began after 45, 65 and 75 days in birds pre-exposed to 8 h light:16 h dark for 4, 6 and 10 weeks, respectively. These results are discussed in relation to other evidence for the progressive development of photosensitivity. PMID- 7473411 TI - Nuclear transfer of putative rabbit embryonic stem cells leads to normal blastocyst development. AB - Rabbit embryonic stem-like cells, characterized by embryoid body formation and differentiation into cell types representative of all three germ layers, were studied for their ability to promote early embryonic development after nuclear transfer. After culture of the reconstructed embryos, 23% (n = 35) developed successfully into morulae or blastocysts, compared with 34% (n = 62) for cloned embryos derived from nuclear transfer with embryonic blastomeres. The cloned embryos from the embryonic stem-like cells appeared normal, with an average of 26% inner cell mass cells, similar to that of control non-manipulated embryos (25%) or cloned embryos from blastomeres (25%). Thus, nuclear transfer of rabbit embryonic stem-like cells leads to early embryonic development that is indistinguishable from blastomere fusion. These results have implications for the development of gene targeting in a species (rabbit) that may be a more suitable model for studying certain human diseases. In addition, this technique may be applicable to other species from which putative embryonic stem cells have been derived, particularly agriculturally important animals. PMID- 7473412 TI - Changes in insulin-like growth factor-binding protein-2 and -3 in follicular fluid during atresia of follicles grown after ovulation in pigs. AB - In pigs, medium-sized follicles (3-6 mm in diameter), lost during preovulatory maturation, are rapidly replenished after ovulation, increasing tenfold in number between day 3 and day 8 of the oestrous cycle. The incidence of atresia in medium sized follicles was previously found to increase between day 5 and day 7 after the onset of oestrus (day 0). This study was conducted to determine whether changes in concentrations of insulin-like growth factor binding proteins (IGFBPs) in pig follicular fluid from medium-sized follicles were related to increased incidence of atresia. Individual follicles were isolated on days 5, 6 and 7 after oestrus from eleven pigs. Follicles were defined as atretic when they contained > or = 10% apoptotic granulosa cells (cells containing subdiploid amounts of DNA fluorescence determined by flow cytometry). Concentrations of IGFBP-2 and -3 were determined by ligand blot analysis with 125I-labelled insulin-like growth factor II and quantified by densitometry. The mean percentage of apoptotic cells per follicle increased (P < or = 0.01) from 3.6 to 35.0 and the mean concentration of IGFBP-2 increased (P < or = 0.001) 3.3-fold between day 5 and day 7 after oestrus. The concentration of IGFBP-3 did not differ significantly among days. Mean oestradiol concentration in follicular fluid decreased (P < or = 0.05) from 52.3 to 0.9 ng ml-1 between day 5 and day 7. Androstenedione concentrations in follicular fluid also decreased (P < or = 0.01) between day 5 and day 7, but progesterone did not differ significantly among days.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7473413 TI - The effect of collection temperature, cooling rate and warming rate on chilling injury and cryopreservation of mouse spermatozoa. AB - The experiments presented here identify several factors that affect survival (motility) of cryopreserved mouse spermatozoa after freezing and thawing. Among these factors are: (i) the temperature at which spermatozoa are collected, (ii) the cooling rate to 0 degrees C and (iii) the warming rate from -196 degrees C to ambient. When excised epididymides were cooled to near 0 degrees (1-4 degrees C) and spermatozoa collected and mixed with cryoprotectant at that temperature, motilities after subsequent freezing and thawing were 8-10 times higher than when the spermatozoa were collected from the epididymides at 22 degrees C. In addition, the survival rates of spermatozoa warmed at rates ranging from 150 to 2000 degrees C min-1 were about five times higher than those in suspensions warmed at about 7500 degrees C min-1. The combination of a low collection temperature and the lower warming rates resulted in approximately 50% motility relative to unfrozen controls. Motility was reduced to 6-8% when the collection temperature was 22 degrees C, and to approximately 10% when frozen suspensions of spermatozoa collected in the cold were rapidly warmed from -196 degrees C. When spermatozoa collected at 22 degrees C were abruptly cooled to 0 degrees C, 40-80% of the cells suffered an irreversible loss of motility after warming. In contrast, when spermatozoa were cooled to 0 degrees C at 1 degree C min-1 and warmed (either rapidly or slowly), motilities were similar to those of uncooled controls (75-90%).(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7473414 TI - In vitro and in vivo evidence on the site of neutralization of equine chorionic gonadotrophin (eCG) by an eCG antiserum. AB - This study was designed to determine whether the major site of eCG neutralization by an antiserum to the hormone is at the peripheral or ovarian level. Hamsters hypophysectomized at oestrus were injected s.c. with 25 iu eCG. Three days later, preovulatory follicles were dissected and cultured for 5 h and the medium was changed every hour. At the end of the first hour of incubation, oestradiol and androstenedione accumulation was high, with a sharp drop over the next 4 h, whereas progesterone concentrations did not change over the entire period. Addition of eCG antiserum to the incubated follicles did not affect steroidogenesis. Addition of 1.0 iu eCG in the second hour or every hour sustained oestradiol production at supraphysiological amounts. However, addition of eCG plus eCG antiserum every hour eliminated the stimulatory effects of eCG on oestradiol production. In another experiment, hamsters injected with eCG were treated 3 days later by i.p. injection of eCG antiserum and groups of animals were killed over the next 8 h. Serum samples before and after injecting eCG antiserum were incubated overnight with a goat anti-rabbit immunoglobulin to separate free, unbound eCG from bound eCG. At time zero (before injecting the antiserum) free eCG was increased, but within 1 h after eCG antiserum there was an eightfold decrease of the hormone, and these concentrations were maintained over the next 7 h. The fall in unbound eCG in vivo coincided with the decay in serum oestradiol and androstenedione.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7473415 TI - Melatonin-binding sites in the gonadotroph-enriched zona tuberalis of ewes. AB - The presence of melatonin-binding sites in the ovine pars tuberalis is well established, but data on melatonin binding in the pars distalis are inconsistent. The distribution of melatonin-binding sites in the ovine hypophysis was investigated using in vitro autoradiography and the high-affinity, high specific activity ligand 2-[125I]iodomelatonin. The histology of sections was visualized with Heidenhain's azan stain and sections were immunoreacted against ovine LH (beta-subunit) using standard immunocytochemical techniques. Melatonin binding in the hypophysis was restricted to the pars tuberalis and the zona tuberalis. The zona tuberalis is histologically similar to the pars tuberalis and appears to be a ventral extension of this region, although the shape and size of the zona tuberalis are extremely variable between individuals. Like the anteroventral pars tuberalis, there is a high concentration of immunoreactive gonadotrophs in the zona tuberalis. The density of immunoreactive gonadotrophs alone is sufficient to discriminate between the zona tuberalis and the pars distalis. Our data suggest that the zona tuberalis and the pars tuberalis are part of the same endocrine tissue and that melatonin-binding sites are not present in the pars distalis proper. PMID- 7473416 TI - Ratio of inner cell mass and trophoblastic cells in demi- and intact pig embryos. AB - Pig morulae, early blastocysts and blastocysts were microsurgically bisected to produce zona-free demi-embryos or remained nonbisected with or without zona pellucida, and the presence of inner cell mass cells was determined using a differential fluorochrome staining technique. After 24 h of in vitro culture, all demi-embryos were classified into three categories, based on morphological criteria: 1, excellent; 2, fair; and 3, degenerated. The average number of total cells and inner cell mass cells in intact embryos cultured without zona pellucida for 24 h was higher (P < 0.05) than that for those with zona pellucida in morulae and early blastocysts. The percentage of demi-embryos without inner cell mass cells in these different morphological categories was 18.7%, 22.2% and 29.8% for morulae, respectively; 3.8%, 16.7% and 30.8% for early blastocysts, respectively; and 3.7%, 32.0% and 36.4% for blastocysts, respectively. The percentage of demi embryos without inner cell mass cells was lower (P < 0.01) in demi-embryos classified in category 1 compared with category 3 in early blastocysts and in category 1 compared with categories 2 and 3 in blastocysts. Significant differences in the total number of cells and the number of inner cell mass cells were apparent among the three morphological categories of demi-embryos derived from morulae, early blastocysts and blastocysts. The ratio of total cells to inner cell mass cells was similar among intact pig embryos and the different morphological categories of demi-embryos derived from morulae, early blastocysts and blastocysts, with the exception of that between demi-blastocysts of category 1 and the other groups.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7473417 TI - Effects of insulin-like growth factor I and interactions with transforming growth factor alpha and LH on proliferation of chicken granulosa cells and production of progesterone in culture. AB - The effects of insulin-like growth factor I (IGF-I), alone, and in combination with LH or transforming growth factor alpha (TGF-alpha), on replication and progesterone production by cultured avian granulosa cells obtained from the three largest (F1-F3) follicles were studied. IGF-I and TGF-alpha stimulated proliferation of granulosa cells in a dose-dependent manner, and responsiveness decreased as the cells matured. IGF-I stimulated progesterone production from granulosa cells of all the follicles with no change in ED50 value during follicular maturation; however, the maximum response was from cells derived from F1 follicles. IGF-I plus LH had an additive effect on progesterone production by cells from all follicles. In contrast, TGF-alpha inhibited basal and LH- and IGF I-stimulated progesterone production. These data show that IGF-I and TGF-alpha may interact with each other during granulosa cell maturation, such that efficacy of IGF-I increases, while that of TGF-alpha decreases before ovulation. Furthermore, both growth factors interact with LH, either to enhance or inhibit progesterone production by granulosa cells. However, LH, IGF-I and TGF-alpha combine to stimulate proliferation of granulosa cells. PMID- 7473418 TI - Effect of testicular damage induced by cryptorchidism on insulin-like growth factor I receptors in rat Sertoli cells. AB - The distribution and density of functional insulin-like growth factor I (IGF-I) receptors in cryptorchid and scrotal rat testes and epididymides during gonadal development were studied. Cryptorchidism was induced by unilateral gubernaculectomy in 4-day-old animals, and organs were studied at 15, 30, 60 and 90 days of age. Tissue membranes were assayed for 125I-labelled IGF-I binding. Characterization and specificity of binding sites showed that both normal and contralateral undescended testes and epididymides exhibited typical type 1 IGF receptors. In normal testes, IGF-I receptor density was 20.6 nmol g-1 wet mass at day 15, and decreased to 12.8 nmol g-1 wet mass at adult age (day 90). Cryptorchid testes showed IGF-I receptor concentrations similar to normal testes at day 15 and day 30, but in postpubertal stages displayed a divergent pattern, with a continuous increase at day 60 and day 90, reaching a higher density than those found for immature ages (62 nmol g-1 wet mass). Both normal and cryptorchid epididymides had a similar concentration and a comparable decrease in IGF-I receptors throughout development. In studies with immunohistochemical techniques (alpha IR-3 antibody), IGF-I receptors were found in primary spermatocytes, Sertoli cells and Leydig cells. Cryptorchid tubules showed a lack of germinal epithelium and a marked increase of immunoreactive IGF-I receptors in Sertoli cells, compared with normal tubules from scrotal testes. Intense immunoreactivity for IGF-I receptors was present in the principal cells of epididymal tubules in both normal and cryptorchid organs.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7473419 TI - Chromatin organization, meiotic status and meiotic competence acquisition in mouse oocytes from cultured ovarian follicles. AB - Changes in chromatin organization, meiotic status and the development of meiotic competence in oocytes retained within mouse ovarian follicles from day 0 to day 6 in culture were examined. The effects of exposure for 24 h to human luteinizing hormone (hLH) during the last day in culture was also determined. Preantral follicles from 22- to 24-day-old (prepubertal) mice develop antra and undergo significant growth from day 0 to day 4 in culture, after which the growth rates slow. The statistical significance of meiotic progression was examined using exact logistical regression analysis, which is particularly useful when the data are sparse and unbalanced. The transition from rimmed to unrimmed germinal vesicle stages was found to occur between day 2 and day 4 of follicle culture and was not influenced by exposure to hLH. Treatment with hLH caused a significant increase in the proportion of intrafollicular oocytes resuming meiosis. Assays of meiotic competence performed in vitro in oocytes retrieved from cultured follicles demonstrated that the transition from an unrimmed to a rimmed state is closely coincident with the acquisition and expression of meiotic competence. Forty-six per cent of competent oocytes from follicle cultures at day 3 progressed to metaphase II. These results indicate that the follicle culture system used in these studies supports the transformation of enclosed oocytes from a precompetent to a competent state and can maintain meiotic arrest for up to 6 days in culture. However, an increasing proportion of oocytes exhibit abnormal meiotic progression with continued follicle culture beyond 4 days. PMID- 7473420 TI - In vitro bioactivity of gonadotrophin surge attenuating factor is not affected by an antibody to human inhibin. AB - The effect of anti-inhibin antibodies on gonadotrophin surge attenuating/inhibiting factor (GnSAF/GnSIF) and its effect on gonadotrophin secretion in a pituitary cell bioassay were determined by culturing rat pituitary cells in a serum-free medium to which inhibin and a partially purified preparation of gonadotrophin surge inhibiting factor (GnSIF) were added. The samples were treated with an anti-inhibin antiserum and basal and GnRH-stimulated FSH and LH secretion were measured by radioimmunoassay of the culture supernatant. Inhibin had a dose-dependent inhibitory effect on both basal and GnRH-stimulated FSH secretion and also on GnRH-stimulated LH secretion. Anti inhibin antibody blocked the inhibition of FSH secretion, except at the highest two doses of human inhibin (1.25 ng ml-1, P = 0.04; 2.5 ng ml-1, P = 0.0004). The GnSIF preparation inhibited basal and GnRH-stimulated FSH and GnRH-stimulated LH secretion and was not affected by the anti-inhibin antibody. The novel hormone, GnSIF, is different from inhibin. PMID- 7473421 TI - Evidence for a local role of inhibin or inhibin alpha subunits in compensatory ovarian hypertrophy. AB - The aim of this study was to determine whether immunoneutralization of inhibin altered compensatory ovarian hypertrophy. Crossbred postpubertal gilts actively immunized with a synthetic bovine inhibin peptide fragment (bINH) conjugated to human alpha globulins (HAG, n = 4 gilts) or HAG alone (control; n = 5) were unilaterally ovariectomized at mid-cycle. After unilateral ovariectomy, the remaining ovary was removed between day 8 and day 12 of the subsequent oestrous cycle. The number of corpora lutea per ovary was determined at each ovariectomy. Blood samples were collected at frequent intervals beginning 1 h before and continuing until the first oestrus after unilateral ovariectomy, and serum concentrations of FSH, LH, progesterone and oestradiol were determined. Inhibin antibody titres were estimated from the percentage of 125I-labelled bINH bound to serum diluted 1:4000. At unilateral ovariectomy, the number of corpora lutea per ovary was similar for bINH:HAG-immunized and control gilts (8.6 +/- 0.7 versus 7.6 +/- 0.6). During the next oestrous cycle after unilateral ovariectomy, the number of corpora lutea on each remaining ovary had doubled (P < 0.05) in controls compared with the number of corpora lutea per ovary in the previous cycle. In contrast, the number of corpora lutea remained unchanged in bINH:HAG immunized gilts. Titre of anti-inhibin antibodies in bINH:HAG-immunized gilts was 9 +/- 1% at unilateral ovariectomy compared with 0% for controls. Alterations in serum concentrations of hormones after unilateral ovariectomy did not differ between treatment groups. Compensatory ovarian hypertrophy was blocked after unilateral ovariectomy in immunized gilts independent of alterations in serum hormones, duration of oestrous cycle, or normal ovulation rate per ovary.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7473422 TI - The presence of water channel proteins in ram and human sperm membranes. AB - Ram and human spermatozoa have a high coefficient of osmotic water permeability (Pf) with a low activation energy (Ea), suggesting the presence of water channels within their plasma membranes. Sperm membranes were examined for the presence of two known water channel proteins, CHIP28 and glucose transporters belonging to the GLUT family of proteins. The water permeability of ram spermatozoa was not inhibited by mercuric chloride to which the CHIP28 channel is sensitive. The CHIP28 protein was not located in western blots of ram sperm membrane preparations that used an anti-CHIP28 antibody. The water permeability of ram and human spermatozoa was inhibited in the presence of phloretin, an inhibitor of glucose transport. Rabbit spermatozoa, which have a low Pf and a high Ea value, suggesting a non-porous membrane, were unaffected by phloretin. These results indicate that the erythrocyte and proximal tubule water channel, CHIP28, is not present in sperm membranes but that sperm membrane glucose transporters may have a secondary water channel function. PMID- 7473423 TI - Functional analysis using chlortetracycline fluorescence and in vitro fertilization of frozen-thawed ejaculated boar spermatozoa incubated in a protein free chemically defined medium. AB - Cumulus-enclosed pig oocytes were matured in vitro, freed from cumulus cells, and inseminated with frozen-thawed ejaculated spermatozoa in a chemically defined protein-free medium containing 37.0 mmol NaHCO3 l-1 and 5 mmol caffeine l-1. When the medium was supplemented with 1 mg polyvinylalcohol (PVA) ml-1, more penetrated oocytes were observed 14 h after insemination with 7-12 x 10(6) cells ml-1 than with 4-5 x 10(6) cells ml-1 and the incidence of polyspermy reflected the sperm concentration used. Varying the NaHCO3 concentration but maintaining the sperm concentration at 8 x 10(6) cells ml-1 resulted in significantly more oocytes being penetrated in media containing 45.83-50.25 than 37.0-41.42 mmol NaHCO3 l-1; there were no significant differences in the incidence of either male pronuclear formation or polyspermy. In medium containing 45.83 mmol NaHCO3 l-1, the inclusion of PVA at 0-5 mg ml-1 had no effect on proportions of penetrated oocytes, male pronuclear formation or polyspermy. However, when spermatozoa from three different boars were evaluated, the penetration and male pronuclear formation rates were highly variable, unlike the incidence of polyspermy. Penetration of cumulus-free oocytes was first detected at 6 h. When spermatozoa were incubated for 6 h in the absence of oocytes, motility, but not vitality, decreased whether or not PVA was included in the medium. Chlortetracycline (CTC) fluorescence analysis of the capacitation state indicated a rapid decline in the proportion of live uncapacitated, acrosome-intact cells and a rapid rise in the proportion of live capacitated, acrosome-reacted cells during the first hour.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7473424 TI - Influence of protein and hormone supplementation on in vitro maturation and fertilization of domestic cat eggs. AB - Immature eggs were recovered from freshly excised ovaries from domestic cats, and initially 931 eggs with a compact cumulus and uniform cytoplasm were cultured in 1 of 15 treatments. Eagle's minimum essential medium containing glutamine and pyruvate was supplemented with 5% (v/v) fetal calf serum (FCS), 4 mg bovine serum albumin (BSA) ml-1 or 2 mg polyvinyl alcohol ml-1 (PVA; non-protein control). Within each of these supplement groups, eggs were cultured with: no hormone; LH+FSH; LH+FSH+oestradiol; or LH+FSH+oestradiol+progesterone. After incubation for 52 h, eggs were inseminated with conspecific fresh spermatozoa, cultured and examined for stage of meiotic maturation and in vitro fertilization (IVF). There were fewer (P < 0.05) eggs maturing to metaphase II in vitro in FCS compared with BSA or PVA, the last two treatments producing similar (P < 0.05) results. Gonadotrophins in concert with oestradiol or oestradiol+progesterone improved the incidence of maturation (P < or = 0.01) compared with no added hormones. The incidence of fertilization and cleavage in vitro ranged from 5.2 to 33.9% and varied (P < 0.05) with hormone subtreatment. Adding FSH+LH+oestradiol consistently increased the incidence of IVF approximately twofold compared with controls with no added hormones. Although it inhibited the ability of eggs to reach metaphase II, FCS in the presence of gonadotrophins and oestradiol allowed > 60% of mature eggs to fertilize in vitro (P < 0.05, compared with PVA and BSA). Inhibitory effects on egg maturation were further evaluated by testing four FCS batches from three commercial sources against BSA.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7473425 TI - Effects of macrophage colony-stimulating factor on folliculogenesis in gonadotrophin-primed immature rats. AB - The effect of macrophage colony-stimulating factor (M-CSF) on folliculogenesis and ovulation was studied. Folliculogenesis and ovulation were induced in immature female rats with a s.c. injection of equine chorionic gonadotrophin (eCG), followed 48 h later by human chorionic gonadotrophin (hCG). The ovulation rate was measured after the following treatments. (1) Graded doses of human M-CSF (1-300 x 10(3) iu per rat) were administered i.p. daily for 3 consecutive days. (2) M-CSF (100 x 10(3) iu) was administered i.p. at designated times between 96 h before and 10 h after hCG injection. (3) Rabbit anti-human M-CSF polyclonal antibody (5 micrograms) was administered into the left ovarian bursa at designated times between 49 h before and 10 h after hCG injection. In addition, the effect of M-CSF on ovarian macrophages was investigated using immunohistochemistry with mouse anti-rat macrophage monoclonal antibody, TRPM-3. The treatment with M-CSF (> 30 x 10(3) iu per rat) significantly increased the ovulation compared with controls in a dose-dependent manner. This stimulatory effect of M-CSF was observed when it was administered between 96 h and 49 h before hCG injection. The ovarian intrabursal administration of anti-M-CSF antibody significantly inhibited the number of ovulated ova from the treated ovaries compared with either those from control rats or from the contralateral untreated ovaries between 24 h before and 3 h after hCG injection. The immunohistochemistry revealed that M-CSF increased the number of ovarian macrophages in growing follicles.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7473426 TI - Ultrastructural and morphometrical analyses of Leydig and Sertoli cells in the testes of rats with hereditary polydactylism. AB - Rats of the polydactylous (PD) strain carry an autosomal recessive gene pd that causes polydactylism in homozygotes (pd/pd). Male homozygotes are sterile owing to an abnormality of spermatogenesis. In the present study, Leydig and Sertoli cells of 12-week-old pd/pd male rats were examined for ultrastructural alterations in an attempt to clarify the cause of the abnormal spermatogenesis. The relative volumes of the organelles were also determined with morphometry. Phenotypically normal pd/+ males served as controls. No morphological or morphometrical abnormalities were noted in the Leydig cells. However, two different types of Sertoli cell were evident: light cells and dark cells. The incidence of the dark Sertoli cells in pd/pd males was high (27%) compared with that in pd/+ males (6%). These dark cells contained quantities of lipid droplets in the cytoplasm and exhibited a significant increase in the relative volume of lipid droplets compared with the value for the light cells. The nuclei of the dark Sertoli cells were irregular in shape and were invaginated. These results suggest that dark Sertoli cells may have lower lipid metabolism, and in pd/pd males, the high number of these dark Sertoli cells may be related to abnormal spermatogenesis. PMID- 7473427 TI - Effect of acute immunoneutralization of inhibin in ewes during the late luteal phase of the oestrous cycle on ovarian hormone secretion and follicular development during the subsequent follicular phase. AB - Ewes with ovarian autotransplants received either inhibin antiserum (10 ml i.v.; n = 6) or sheep serum (10 ml i.v.; n = 5) on day 10 of the luteal phase with additional daily injections (1 ml i.v.) from 48 h after the initial injection until the end of blood sampling, 9 days later. Luteal regression was induced by injection of prostaglandin F2 alpha (PGF2 alpha) 4.2 days after the initial plasma injection. Jugular and ovarian venous blood samples were taken every 4 h over the experimental period, and more frequent samples (every 10-15 min for 2-3 h) were taken to examine pulsatile secretory responses from the ovary to GnRH induced (150 ng i.m. 1 and 4 days after initial treatment) or endogenous LH pulses (24 and 48 h after injection of PGF2 alpha). Plasma FSH concentrations, ovarian steroid secretion and ovarian follicular development were measured. Immunization against inhibin resulted in a two- to threefold increase (P < 0.001) in plasma FSH concentrations, which remained higher than controls until injection of PGF2 alpha. Within 24 h of immunization, there was an increase in the number of small ovarian follicles (P < 0.01) and by 4 days after treatment, immunized ewes had four or five (P < 0.01) large ovarian follicles and, despite little change in the basal steroid secretion, a four- to sixfold increase (P < 0.05) in the amplitude of the steroidogenic response to a GnRH-induced LH pulse.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7473428 TI - Analysis of sperm quality in recombinant inbred mouse strains: correlation of sperm head shape with sperm abnormalities and with the incidence of supplementary spermatozoa in the perivitelline space. AB - Recombinant inbred strains were developed from reciprocal crosses between two inbred strains of mice (CBA and KE) differing in sperm head shape, proportion of normal sperm heads (CBA, 95%; KE, 78%) and fertilization efficiency (CBA, 100% of fertilized ova; KE, 72%), to determine whether the indices of sperm morphology and function were correlated. The following parameters were analysed in recombinant inbred and progenitor strains: index of sperm head shape (head width in the middle of its length/head length), percentage of abnormal sperm heads, percentage of spermatozoa with progressive movements, efficiency of penetration of hyaluronic acid polymer (Sperm Select) and percentage of fertilized ova after mating males from the tested strains with females from an outbred stock. For each investigated character, recombinant inbred strains, recombinant inbred EXCB and CBXE, could be divided into at least three categories: KE-like, CBA-like and intermediate, suggesting that in each case a minimum of two genes was involved. Recombinant strains derived from the reciprocal crosses of progenitor strains differed only with respect to the proportion of abnormal sperm heads, showing the involvement of the Y chromosome in determining this character. Penetration into Sperm Select was significantly correlated both with fertilization efficiency and sperm motility, while correlation with the proportion of normal spermatozoa did not reach the level of significance. However, there was a significant negative correlation of both sperm abnormalities and the incidence of supplementary spermatozoa in the perivitelline space with the index of sperm head shape.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7473429 TI - Intravenous immunoglobulin infusion therapy in women with recurrent spontaneous abortions of immune etiologies. AB - We have investigated clinical effectiveness of intravenous immunoglobulin G infusion (IVIg) on antiphospholipid antibody titers in five women with evidence of antiphospholipid antibody-associated recurrent spontaneous abortions and one with antinuclear antibody who became refractory to conventional autoimmune treatment during pregnancy and experienced pregnancy complications. Three women developed intrauterine growth retardation and three had complicated twin pregnancies with rising autoantibody titers. Antiphospholipid antibody and antinuclear antibody titers were tested pre and 2 weeks after each IVIg infusion. We report that: (i) IgG antiphospholipid antibody titers were significantly suppressed after each IVIg infusion (P < 0.05); (ii) IgM antiphospholipid antibody titers were also significantly suppressed after each IVIg infusion (P < 0.0001); (iii) decreased titers of autoantibodies paralleled increased levels of maternal IgG which lasted for at least 30 days; the autoantibodies showed a definite rise again prior to the next infusion; (iv) antinuclear antibody titers were effectively suppressed; and (v) rising autoantibody titers combined clinical manifestation of intrauterine growth retardation and women with complicated twin pregnancies. We conclude that IVIg infusion effectively suppresses IgM and IgG autoantibodies to phospholipids and antinuclear antibody in autoimmune women with a history of recurrent spontaneous abortions and refractory to conventional anticoagulation or immunosuppressive treatment. PMID- 7473430 TI - Indomethacin blocks the immunosuppressive activity of rat testicular macrophages cultured in vitro. AB - Macrophages are a prominent resident cell type in the interstitial tissue of the testis in several mammalian species. This presence in an immunologically privileged site prompted an investigation of their ability to initiate and regulate lymphocyte proliferation in vitro. Isolated rat peripheral blood lymphocytes (PBL) were cultured either directly with isolated rat testicular (Tm) or peritoneal (Pm) macrophages or with the conditioned medium from cultures of these cells (Ts or Ps). The presence of Tm and Ts reduced the proliferative response of PBL to 65% +/- 3% and 65% +/- 4% of that observed in the control cultures. Stimulation of the Tm with lipopolysaccharide (LPS) did not significantly alter this effect. Dialysis of Ts (to remove molecules < 14,000 MW) before addition to PBL cultures did significantly reduce the amount of inhibition, with PBL proliferation reaching 93% +/- 4% of the control. LPS in conjunction with indomethacin (IDM) or interferon gamma (IFN gamma) induced PBL proliferation at levels comparable to or significantly greater than those of the controls (104% +/- 4% and 113% +/- 6%, respectively). The collective addition of IDM, IFN gamma and LPS to Tm cultures increased PBL proliferation over control levels (119% +/- 5% for Ts and 133% +/- 6% for Tm). Prostaglandin levels in macrophage-conditioned medium were measured by specific radioimmunoassay and were significantly greater in Ts (13.1 +/- 0.4 ng/ml PGE2 and 16.8 +/- 0.6 ng/ml PGF2 alpha) than in Ps (both below the assay minimum sensitivities). The results indicate that the rat testicular macrophages produce high basal levels of prostaglandin E2 and F2 alpha. These products appear largely responsible for the inhibition by these cells of mitogen-induced PBL proliferation in vitro, and may contribute to both the immune-privileged status and the normal physiology of the rodent testis. PMID- 7473431 TI - Evidence that secretory products of the reticuloepithelial cells of the rat thymus modulate the secretion of gonadotrophins by rat pituitary cells in culture. AB - It is now well accepted that the endocrine system can be strongly influenced by the immunological status of the body. This laboratory has provided documented evidence that the rat thymus produces a biologically active secretion, with a molecular weight of approximately 30 kDa, that augments the gonadotrophin releasing hormone (GnRH) stimulated secretion of gonadotrophins by isolated pituitary cells in culture. To determine the cells of origin of the factor involved, the reticulo-epithelial cells from the thymus were incubated in vitro to provide media for the examination of possible effects on the secretion of gonadotrophins by the rat pituitary. The enrichment of the culture with reticulo epithelial cells was confirmed using anticytokeratins antibodies. It was found that the addition of such conditioned medium increased the secretion of luteinizing hormone (LH) and follicle stimulating hormone (FSH), with a greater effect on the former. The increment in gonadotrophin secretion was time- and dose dependent. This same conditioned medium also potentiated the effect of GnRH on the cultured pituitary cells. The fractionation of thymic-conditioned medium using selective molecular membrane filters, provided evidence that the active factor or factors involved in the enhanced secretion of gonadotrophins are larger than 10 kDa MW. Present findings show that thymic reticulo-epithelial cells are responsible for this activity. PMID- 7473432 TI - Phenotypical heterogeneity of testicular macrophages/dendritic cells in normal adult mice: an immunohistochemical study. AB - The distribution of macrophage/dendritic cell antigens was investigated immunohistochemically in frozen testis sections of normal A/J mice using a panel of rat monoclonal antibodies against murine macrophage/dendritic cell antigens (F4/80, BM8, MP23, MOMA1, MOMA2, M5/114, BMDM1 and NLDC145). Sections of lymphoid organs were employed as positive controls. Stained-macrophages/dendritic cells in the testicular interstitium were various in shape, such as irregular-, elongate-, round- or oval-shaped cytoplasm with short or long cytoplasmic protrusions, but, the most conspicuous morphology was irregular. The testicular macrophages/dendritic cells were subdivided into four types of cells by their localization; various-shaped cells associated with Leydig cells and other interstitial cells (Type I); stretched cells attached to the seminiferous tubular wall (Type II); stretched cells attached to the blood vessel wall (Type III); and stretched cells attached to the inner side of the testicular capsule (Type IV). Examination of the staining pattern of the various antibodies showed that the macrophage/dendritic cell antigens differed from each other in amount present and localization, indicating phenotypical heterogeneity of the testicular macrophages. Additionally, a few macrophages were identified in the testicular capsule and in the tubuli recti but never in the seminiferous tubules. PMID- 7473433 TI - Expression of endogenous HIV-1 crossreactive antigens within normal human extravillous trophoblast cells. AB - Expression of intact endogenous retroviruses by normal placental villous trophoblast and immuno-crossreactivity of villous trophoblast with anti retroviral antisera have been documented. The nature and/or potential function of these particles/proteins has not yet been fully defined. We previously reported that monoclonal antibodies directed against HIV-1 envelope and gag proteins react with normal human villous trophoblast. In this study, we report that extravillous trophoblast (EVT) from second- and third-trimester tissue are also cross-reactive with anti-HIV-1 gp120/160 and p17/18 antibodies. We document a differential expression of such cross-reactive epitopes between mononuclear EVT and placental bed giant cells. Mononuclear EVT principally displayed reactivity throughout the cytoplasm with little or no difference between cells, whereas placental bed giant cells displayed distinct localization of labeling to limited areas of cytoplasm. This pattern of reactivity apparently correlates with trophoblast morphological differentiation and with our earlier observations concerning villous trophoblast. These data illustrate that retrovirus-associated epitopes are expressed by trophoblast throughout the normal human placenta and that this distribution is related to morphologic differentiation of these cells. PMID- 7473434 TI - Systemic and mucosal antibody responses to group B streptococci following immunization of the colonic-rectal mucosa. AB - The cervico-vaginal mucosa is poorly designed for inducing a mucosal immune response, but it can effect such a response evoked at other mucosal sites. This study was undertaken to determine whether colonic-rectal immunization with group B streptococci (GBS) might induce a local cervico-vaginal immune response. Mice were immunized with either fragmented GBS rectally, whole GBS rectally, or whole GBS subcutaneously. Cholera toxin (CT) was used as an adjuvant for the rectal immunizations. Following colonic-rectal immunization with whole GBS, the mean anti-GBS IgA antibody level in vaginal secretions was 735 kU/ml, with individual values reaching 3480 kU/ml. Corresponding levels of IgA antibodies never exceeded 10 kU/ml in serum and intestinal secretions, or 90 kU/g in feces. In vaginal secretions IgA antibodies to GBS also constituted a much larger fraction of total IgA than in serum, intestinal secretions and feces. Immunizations with fragmented GBS produced much lower IgA responses. Anti-GBS IgA response at the inductive site in the colon-rectum was not significant, as opposed to a strong anti-CT IgA response. Except in serum, the anti-GBS IgG responses to colonic-rectal immunizations were generally low, or absent. The results may provide a basis for the development of mucosal vaccines against GBS-infection. PMID- 7473435 TI - Effects of testicular cytokines on proliferation of rat T-leukaemic lymphoblasts in vitro. AB - A rat acute lymphoblastic leukaemia (ALL) model was used to study the mechanisms involved in the tendency to testicular relapse of ALL in boys. Previous studies have indicated that the infiltration and growth of leukaemic lymphoblasts in the testis are influenced by the same endocrine and paracrine control systems that regulate normal testicular function. In the present study the effects of aqueous extracts of scrotal, abdominal and estrogen-treated postpubertal rat testes on rat-leukaemic lymphoblast proliferation were evaluated. The effects of recombinant cytokines analogous to those observed in the testis on leukaemic cell DNA-synthesis were also evaluated since changes in the levels of these factors have been observed in association with cryptorchidism and low levels of gonadotropins. Transforming growth factor-beta 1 (TGF-beta1), significantly inhibited the proliferation of leukaemic rat lymphoblasts after 24 h of culture, whereas TGF-beta 2, interleukin-1 alpha (IL-1 alpha), IL-1 beta, IL-6 or combinations of them were inactive. Extracts of estrogen-treated testes and abdominal testes of unilaterally cryptorchid animals inhibited leukaemic T-cell proliferation significantly more than extracts of normal testes. The inhibitory activity in abdominal testes could be neutralized by anti-TGF-beta 1 antibodies. These results suggest that testicular TGF-beta 1 may influence growth of leukaemic lymphoblasts in the testis but also that other as yet unknown, testicular factors are involved in the regulation of leukaemic cell function in the testis. PMID- 7473436 TI - Surgical management of benign cystic teratoma. Laparoscopy vs. laparotomy. AB - OBJECTIVE: To compare laparoscopy to laparotomy for the surgical treatment of benign cystic teratomas. STUDY DESIGN: This was a retrospective, case series comparison of 20 patients who had surgery at Rochester General Hospital from June 1991 to January 1993 for benign cystic teratomas. Statistical comparisons were made by Student's t test or chi 2 analysis. RESULTS: Eight patients had surgery via laparoscopy and 12 via laparotomy. Laparoscopic oophorectomy and cystectomy resulted in significantly shorter hospital stays and decreased hospital costs, but the surgery time was significantly increased. The patients' fertility status influenced the choice of conservative cystectomy or nonconservative oophorectomy as the surgical procedure. There were no serious complications in any patients, including those with laparoscopic cystectomies and intraperitoneal spill. CONCLUSION: Oophorectomy and ovarian cystectomy via operative laparoscopy appear to be reasonable options for the surgical treatment of benign cystic teratoma. PMID- 7473437 TI - Chronic pelvic pain in adolescent women. Diagnostic laparoscopy and ultrasonography. AB - OBJECTIVE: To evaluate the correlation of preoperative pelvic examination and ultrasonography with the laparoscopic findings in 45 adolescent women with chronic pelvic pain. STUDY DESIGN: Between January 1, 1989, and December 31, 1992, 45 adolescent women underwent diagnostic laparoscopy following a detailed clinical workup for evaluation of chronic pelvic pain. RESULTS: Thirty-five of 45 pelvic examination findings correlated with the laparoscopic findings. The predictive values of normal and abnormal findings at pelvic examination were 42.8% and 93.5%, respectively. Ultrasonography (US) correlated with laparoscopy in 39 of 45 cases. The predictive value of normal findings at US was 60%, and that of abnormal findings was 94.2%. When both pelvic examination and US were normal, 50% of cases were abnormal at laparoscopy. However, laparoscopy revealed abnormality in 100% of patients with abnormal pelvic examinations and abnormal US findings. CONCLUSION: Clinical evaluation of chronic pelvic pain, when combined with US, is highly predictive but needs confirmation, best provided by laparoscopy. PMID- 7473438 TI - Genuine stress urinary incontinence with low urethral pressure. Five-year follow up after the Ball-Burch procedure. AB - OBJECTIVE: To evaluate long-term results of the Ball-Burch procedure in women with genuine stress incontinence (GSI), low urethral pressure type. STUDY DESIGN: Sixty women with a clinical and urodynamic diagnosis of GSI, low urethral pressure type, underwent the Ball-Burch procedure between May 1987 and February 1988. They were evaluated preoperatively and one and five years after surgery. The urogynecologic workup consisted of a history, physical examination, urine culture, neurologic examination, urethrocystoscopy, cotton swab test, urethrocystometry, urethral profilometry and uroflowmetry. Forty-eight women were available at the one-year follow-up, and 32 of the 48 (66.7%) were available after five years; the mean age at the time of surgery was 58.7 years and mean parity, 2. Twenty-seven women were premenopausal and 33 postmenopausal; at the five-year follow-up all but 2 were postmenopausal. All postmenopausal women received estrogen replacement therapy prior to urodynamic evaluation. Cure was defined as absence of loss of urine by history (subjective) and during urodynamic evaluation (objective). The t test and paired t test were used for statistical analysis. RESULTS: Forty-two of 48 women (87.5%) were cured after one year, and 27 of 32 (84%) were cured after five. There was a statistically significant difference between the preoperative and five-year follow-up values for the swab test, maximum urethral closure pressure and pressure transmission ratio at the proximal and mid urethra. Twenty-six of 29 patients with a negative swab test at the one-year follow-up were cured after five years, resulting in a predictive value of 89% for cure after a negative test. CONCLUSION: In our series the Ball Burch procedure was an effective surgical treatment for patients with GSI and low urethral pressure. The swab test, when negative one year after surgery, was predictive of a successful long-term outcome. PMID- 7473439 TI - Alternative prenatal care. Impact of reduced visit frequency, focused visits and continuity of care. AB - OBJECTIVE: To investigate the impact of an alternative prenatal care program for low-risk patients. STUDY DESIGN: Five hundred forty-nine low-risk pregnant women were allocated to the study and control groups. The study group received, on average, eight visits, all of them with one of nine study providers. Each study visit had specific objectives and accompanying targeted patient education handouts. The control group received the usual prenatal care (on average, 13 visits) with different providers, according to the customary schedule. RESULTS: There were no significant pregnancy outcome differences between the groups. The study vs. control group differed significantly (P < .0001) in patient satisfaction regarding the number of prenatal visits. There was a higher level of satisfaction in the study group concerning continuity of care (P < .0001). The alternative prenatal care program reduced the number of prenatal visits by 27% and was not associated with any change in maternal or perinatal outcomes. Patient satisfaction parameters were either maintained or improved with alternative prenatal care. CONCLUSION: An alternative prenatal care program for low-risk patients reduced resource utilization without adversely affecting prenatal care process variables, pregnancy outcome or patient satisfaction. PMID- 7473440 TI - Vibratory acoustic stimulation and fetal gasping. AB - OBJECTIVE: Vibratory acoustic stimulation (VAS) has been used to shorten the duration of antepartum fetal heart rate monitoring. Questions have been asked regarding the possible effects of VAS on the fetus. VAS was used to investigate whether the acute effect of VAS is associated with gasping in the normal fetus. STUDY DESIGN: Thirty-seven normal patients at 37-42 weeks' gestation with singleton fetuses were studied. All had normal amniotic fluid volume on ultrasound. These 37 subjects had a five-minute baseline sonographic assessment of fetal breathing and body movements. A fetal gasp was defined as an isolated, slow, irregularly occurring, deep, inspiratory-type movement, or series of them, of the chest and abdomen. Three-second VAS was then delivered with an artificial larynx placed directly over the fetal head. Fetal breathing, body and gasping movements were assessed at the time of this stimulus. RESULTS: A significant increase in fetal body movements (P < .001), a significant decrease in fetal breathing (P < .003) and absence of gasping movements were noted. CONCLUSION: The results suggest that gasping does not occur in response to VAS in the normal, term fetus. PMID- 7473441 TI - Alternatives in plasma gonadotropin and sex steroid levels in obese ovulatory and chronically anovulatory women. AB - OBJECTIVE: To evaluate the alterations in plasma gonadotropin and sex steroid levels in obesity. STUDY DESIGN: A study was conducted on 77 obese women, 30 with ovulatory menstrual cycles (group I) and 47 with chronic anovulation (group II), who were compared to 15 women of normal weight and with ovulatory cycles (group C). Obesity was determined on the basis of body mass index. Blood samples were obtained during the follicular phase of the cycle for measurement of follicle stimulating hormone (FSH), luteinizing hormone (LH), prolactin (PRL), testosterone (T), androstenedione (A), dehydroepiandrosterone sulfate (DHEAS), estrone (E1) and estradiol (E2). RESULTS: Elevated LH, T, A and E1 levels were detected in group II when compared to group I and group C. A levels were also elevated (P < or = .05) in group I when compared to group C. There was no significant difference between the three groups in FSH, PRL, DHEAS or E2 levels. CONCLUSION: The elevated A levels in group I obese women show that even when ovulating, obese women are hyperandrogenic. This excessive A may activate the aromatizing system in adipose tissue, generating E1. When this occurs, the increase in E1 may trigger an increase in LH, with consequent ovarian hyperstimulation, leading to increased T and anovulatory cycles. PMID- 7473442 TI - Efficacy of oral micronized progesterone in the treatment of luteal phase defects. AB - OBJECTIVE: Vaginal progesterone suppositories are an accepted treatment for infertility attributed to luteal phase defects. Although oral micronized progesterone may be preferable to suppositories for many patients, there are no studies on its use for patients with luteal phase defects. This study evaluated the efficacy of oral micronized progesterone for the treatment of luteal phase defects. STUDY DESIGN: Seven women with luteal phase defects previously corrected by vaginal suppositories were administered oral micronized progesterone (200 mg by mouth three times a day). Endometrial biopsies were performed to evaluate treatment efficacy. Questionnaires were used to assess side effects, including sedation. RESULTS: On oral micronized progesterone, all patients had in-phase endometrial biopsies. Despite complaints of drowsiness, the majority of patients preferred the oral formulation over the vaginal route of administration. CONCLUSION: We conclude that oral micronized progesterone is efficacious in the treatment of luteal phase defects. PMID- 7473443 TI - Suspected ectopic pregnancy. What to do when human chorionic gonadotropin levels are below the discriminatory zone. AB - OBJECTIVE: To assess the value of a noninvasive diagnostic strategy in cases of suspected ectopic pregnancy when transvaginal sonography (TVS) and serum human chorionic gonadotropin (hCG) measurement show inconclusive findings. STUDY DESIGN: Prospective study of 85 patients without pregnancy detectable on initial TVS together with low hCG levels (< 1,500 IU/L). The diagnostic strategy combined the results of TVS, hCG measurements and time, according to a strict algorithm. RESULTS: Overall test results of the strategy were excellent: sensitivity, 0.94; specificity, 1.00; likelihood ratio of a positive test result, infinity; likelihood ratio of a negative test result, 0.06. The strategy revealed a new diagnostic entity, trophoblast in regression (TIR). TIR represents patients with declining hCG levels and unknown locations of their pregnancies, presenting a benign clinical picture, and constituted 54% of the population studied. CONCLUSION: This noninvasive diagnostic strategy proved to be very reliable. Because the strategy integrated expectant management of patients with declining hCG levels, it prevented many unnecessary surgical procedures. This was of great benefit for both patients with early intrauterine pregnancies and those with TIR. PMID- 7473444 TI - Ability of visual tests to predict underlying cervical neoplasia. Colposcopy and speculoscopy. AB - OBJECTIVE: To describe the attributes of colposcopy and a low-power, magnified examination that utilizes chemiluminescent illumination (speculoscopy) in the visualization of cervical epithelium in a predefined, high-risk population and to compare how the two tests predict cervical histology. STUDY DESIGN: During this multicenter, prospective study, 395 women who were referred to our colposcopy clinic underwent a repeat cervical smear and speculoscopy followed immediately by colposcopy. Abnormal colposcopic lesions were biopsied and endocervical curettage performed when indicated. Histologic diagnoses were compared with cytology, speculoscopy and colposcopy results. RESULTS: Colposcopy was more sensitive than speculoscopy in the detection of cervical neoplasia (97% vs. 82%) (P < .001) and was superior in visualizing focal lesions and vascular patterns. An antecedent acetowhite abnormality detected during speculoscopy was highly predictive of subsequent abnormal colposcopy (97% positive predictive value). The "overall" rate of acetowhite lesions during speculoscopy was nearly half the rate during colposcopy (P < .001). CONCLUSION: Colposcopy is better suited than speculoscopy to the follow-up of patients with abnormal cervical cytology because it facilitates lesion grading and assists in directing biopsies. Speculoscopy is best utilized as a dichotomous screening test based on the presence or absence of at least one well-demarcated acetowhite lesion and may be more suitable than colposcopy as an adjunct to cervical cancer screening due to its lower overcall rate. The biophysical properties of blue-white chemiluminescent light as it relates to the diagnosis of cervical neoplasia are discussed. PMID- 7473445 TI - Prenatal diagnosis of partial trisomy through in situ hybridization on amniocytes with whole chromosome and centromere-specific DNA probes. A case report. AB - BACKGROUND: DNA probes specific for whole chromosomes or portions of chromosomes can provide important information to aid the clinician in managing pregnancy and the geneticist in relaying accurate recurrence risk information to the patient. CASE: In this case, sonography was ordered because of a low fundal height in a 29 year-old primigravida at 35 weeks' gestational age; it revealed major fetal anomalies. A small supernumerary marker was seen in some cultured amniocytes. Metaphase spreads were analyzed by means of fluorescence in situ hybridization using a centromere probe specific for chromosome 22 and a whole chromosome probe for the 11 chromosome. In situ hybridization showed that the marker chromosome was a derivative of chromosome 22 with 11q material attached near the centromere. The fetal karyotype was 47,XY,+der(22) t(11;22)(q23.3;q11.2)mat. The mother was later found to be a balanced translocation carrier. CONCLUSION: It was possible to offer rapid prenatal diagnosis for this family using interphase analysis with the 22 centromere probe. The patient had chorionic villus sampling, and DNA probes were used to analyze cells directly from the biopsy. The signal representing the supernumerary marker was not observed. Karyotype analysis later showed that the fetus was normal but a translocation carrier. This report illustrates that rapid in situ hybridization can provide important information in known cases of translocation carriers. PMID- 7473446 TI - Endometriosis with a pleural effusion and ascites. Report of a case treated with nafarelin acetate. AB - BACKGROUND: Although endometriosis is generally confined to the pelvis, it may occur at remote sites with unusual manifestations. Rare examples include pulmonary endometriosis and endometriosis associated with ascites. These complications represent diagnostic and therapeutic dilemmas, especially when both occur concurrently in the same patient. CASE: A 21-year-old nulligravida had a history of endometriosis and complained of severe dyspnea and weight loss. Pleural effusion and pelvic ascites associated with recurrent endometriosis were found. The patient was treated with thoracentesis and nafarelin acetate nasal spray. A remarkable improvement in her condition occurred, and the side effects of the spray were minimal during treatment. CONCLUSION: Gonadotropin-releasing hormone agonist therapy is recommended as the initial treatment for endometriotic ascites and/or pleural effusion. PMID- 7473447 TI - The Zavanelli maneuver for fetal shoulder dystocia. Three cases with poor outcomes. AB - BACKGROUND: There are several reports on the management of severe fetal shoulder dystocia by pushing the head back into the vagina and delivering the infant by cesarean section; this procedure is called the Zavanelli maneuver. CASES: Three cases were reviewed to determine the outcomes of the infants. The three neonates all demonstrated severe perinatal hypoxia that ultimately resulted in brain damage and/or death. CONCLUSION: Management of shoulder dystocia using the Zavanelli maneuver carries a high risk of severe infant perinatal hypoxia and damage. PMID- 7473448 TI - Abnormal uterine bleeding. Accuracy of vaginal probe ultrasound in predicting abnormal hysteroscopic findings. AB - OBJECTIVE: To determine the accuracy of vaginosonography in predicting the presence of abnormal intrauterine findings at hysteroscopy in women presenting with abnormal uterine bleeding. STUDY DESIGN: Two hundred thirty-eight women between the ages of 25 and 75 were evaluated for abnormal uterine bleeding by a combination of vaginal probe ultrasound, hysteroscopy and suction curettage. Ultrasound findings were classified as normal, abnormal or equivocal, and hysteroscopic findings as normal or abnormal. The ability of findings on ultrasound to predict a normal or abnormal hysteroscopic examination was evaluated. RESULTS: Ultrasound examination was normal, equivocal and abnormal in 57 (24%), 78 (33%) and 103 (43%) patients, respectively. If equivocal scans are excluded, ultrasound had a positive predictive value of 87%, negative predictive value of 89%, sensitivity of 94% and specificity of 89%. If equivocal scans are considered abnormal, the positive predictive value of an abnormal ultrasound examination was 75%, negative predictive value was 89%, sensitivity 96% and specificity 53%. Vaginosonography detected 99% of submucous myomas and 89% of endometrial polyps. CONCLUSION: Normal and abnormal vaginal probe ultrasound examination of the uterus is effective in predicting the presence or absence of endometrial pathology as determined by hysteroscopy and suction curettage. Vaginal probe ultrasound is an excellent screening examination for the presence of intrauterine pathology and can assist in planning further evaluation and treatment. PMID- 7473449 TI - Unipolar electrosurgery in operative laparoscopy. Capacitance as a potential source of injury. AB - OBJECTIVE: To determine whether capacitive "stray" currents that can be generated during unipolar laparoscopic electrosurgery are likely to cause unintentional organ injury. STUDY DESIGN: In a controlled laboratory setting, the passage of capacitive current across the insulated portion of unipolar, 5-mm laparoscopic instruments was assessed for its ability to cause damage to biologic tissues. RESULTS: Modulated (coagulating) current output at power settings as low as 20-25 W led to apparent passage of current across the insulated portion of some laparoscopic instruments with thin insulation. This was accompanied by thermal damage to biologic tissues and instrument insulation. CONCLUSION: Our findings can explain unintentional injury during unipolar electrosurgery. The risk of such injury can be reduced by surgeon awareness, low generator output power and use of instruments with ample insulation. Thin insulation is more likely to be encountered with disposable than with multiuse instruments. PMID- 7473450 TI - Pipelle endometrial sampling. Sensitivity in the detection of endometrial cancer. AB - OBJECTIVE: To evaluate the sensitivity of the Pipelle endometrial suction curette in the detection of endometrial carcinoma in patients with known endometrial cancer. STUDY DESIGN: A study was conducted using patients with known endometrial cancer undergoing a hysterectomy as part of a formal staging procedure. Endometrial biopsies were performed prior to each surgical procedure. Biopsy results were compared to the hysterectomy specimen for specimen adequacy and final histologic diagnosis. RESULTS: The Pipelle biopsy was adequate for analysis in 63 of 65 patients (97%). Malignancy was detected by biopsy in 54 of 65 patients, for a sensitivity of 83 +/- 5% (mean +/- SD). Of the 11 patients with false negative results, 5 had tumors present in only an endometrial polyp. Three of the 11 patients had disease localized to < 5% of the surface area of the endometrium. CONCLUSION: The Pipelle endometrial suction curette is an effective office device for evaluating patients at risk of endometrial cancer; however, tumors localized to a polyp or small area of endometrium may go undetected. PMID- 7473451 TI - Circulating hormone levels in menopausal women receiving different hormone replacement therapy regimens. A comparison. AB - OBJECTIVE: To measure and compare plasma levels of sex hormones after the administration of different hormone replacement therapy (HRT) regimens. STUDY DESIGN: Ninety women with natural menopause were randomized into this comparative study. Eighty-five women completed one year of follow-up. Patients were randomly assigned to five groups. The first received 0.6 mg/d of conjugated equine estrogen (CEE) cyclically (n = 15). The second received 50 micrograms/d of transdermal estradiol (E2) cyclically (n = 17), and the third received 0.6 mg/d of CEE continuously (n = 17). All these groups also received 2.5 mg of medroxyprogesterone acetate (MPA) sequentially for the last 12 days of HRT, while the fourth therapy group received 0.625 mg/d of CEE and 2.5 mg/d of MPA continuously (n = 19). The fifth group constituted a treatment-free control group (n = 22). Levels of follicle-stimulating hormone (FSH), luteinizing hormone (LH), E2, estrone (E1), prolactin (PRL), testosterone (T), androstenedione (A4), dehydroepiandrosterone sulfate (DHEA-S) and sex hormone binding globulin (SHBG) were determined prior to HRT and during the last week of the 6th and 12th months of HRT, between days 21 and 24 of estrogen administration. RESULTS: After HRT we found decreases in FSH, LH and PRL levels, increases in E2, E1 and SHBG, and no modifications in T, A4 and DHEA-S plasma levels. There were no significant differences between the treatment groups in FSH, LH, E2, PRL, T, A4 or DHEA-S. E1 and SHBG were significantly higher in the groups with oral HRT. CONCLUSION: All the observed changes in hormone levels are to be expected after HRT except for the decrease in PRL levels. Finally, although MPA dosage was not the focus of the present study, our results suggest that the dosage of 2.5 mg/d of MPA in sequential regimens is clearly inadequate to protect the endometrium from hyperplastic changes. PMID- 7473452 TI - Intrapartum amniotic fluid index for predicting fetal acidemia. Results of receiver operating characteristic curve analysis. AB - OBJECTIVE: The objective of this study was to determine if the discriminatory intrapartum amniotic fluid index can reliably predict respiratory or metabolic acidosis in the newborn. STUDY DESIGN: Among 256 parturients at 37 weeks' gestation or more, the intrapartum amniotic fluid index was obtained in early labor, and umbilical arterial blood gas analysis was performed within 30 minutes of delivery. The mean (+/- SD) amniotic fluid indices in labor among newborns with no acidosis versus those with respiratory or metabolic acidosis were compared. A receiver operating characteristic curve was used to determine the best discriminatory intrapartum amniotic fluid index for respiratory and metabolic acidosis. RESULTS: Statistical analysis revealed that the mean intrapartum amniotic fluid index among 12 newborns with respiratory acidosis and 26 with metabolic acidosis was significantly lower than the mean index in 218 newborns without fetal acidemia. Despite the differences in the mean intrapartum amniotic fluid index in the three groups analyzed, the receiver operating characteristic curve indicated that an intrapartum amniotic fluid index alone is a poor test for predicting acidosis (respiratory or metabolic) in the neonate. CONCLUSION: The mean intrapartum amniotic fluid index in labor among newborns with respiratory or metabolic acidosis was significantly lower than that among nonacidotic newborns. The receiver operating characteristic curve indicated that the intrapartum amniotic fluid index cannot reliably distinguish nonacidotic infants from those with respiratory or metabolic acidosis. PMID- 7473453 TI - Ultrasonographic characteristics of first-trimester gestations in recurrent spontaneous aborters. AB - OBJECTIVE: To compile, for the first time, serial ultrasonographic findings during the first trimester of pregnancy in women with a history of primary recurrent spontaneous abortion so as to define the dynamics of early normal and abnormal gestations in this category of gravidas. STUDY DESIGN: Transvaginal ultrasonograms were obtained weekly from 5 to 12 weeks' gestational age in 40 women, 10 each of four groups: recurrent spontaneous aborters and primiparas (controls), with both successful and failed gestations. RESULTS: Embryonic heart motion was detected in 40-50% of successful pregnancies during the fifth week of gestation and in the balance by the sixth week, while heart motion was detected in no more than 50% of pregnancies that later failed. Of the failed pregnancies, all were evident by the eighth week of gestation, including those with previously documented viability. The gestational sac size and crown-rump length were smaller than expected in both failed groups, with the sac size difference evident as early as week 5 and the crown-rump length difference apparent by week 7. CONCLUSION: Appropriate timing of the initial ultrasonogram in recurrent aborters (i.e., 8 weeks' gestational age) can identify, by means of heart motion and gestational sac features, all pregnancies that will ultimately fail. PMID- 7473454 TI - Racial and ethnic correlates of tobacco, alcohol and illicit drug use in a pregnant population. AB - OBJECTIVE: To compare risk factors for tobacco, alcohol and illicit drug use prior to entry into prenatal care and lifetime illicit drug use in a large sample of pregnant women stratified by race/ethnicity. STUDY DESIGN: University clinic based study comparing risk factors for substance use among 899 white, black and Mexican-American patients, aged 12-41 years. Crude and adjusted risk factors and 95% confidence limits were calculated for substance use prior to the first prenatal visit and life-time illicit drug use. RESULTS: Prior physical assault placed patients from each racial/ethnic group at risk of using all substances during pregnancy and of lifetime illicit drug use. Other common risk factors included partner's use of alcohol or illicit drugs and previous pregnancy. Risk factors that varied by racial/ethnic group included school enrollment and single marital status. CONCLUSION: Effective interventions will require that practitioners be sensitive to the unique risk factors for substance use among women of different cultural backgrounds. PMID- 7473455 TI - Histologic effects of different suture materials in microsurgical anastomosis of the rat uterine horn. AB - OBJECTIVE: To evaluate the histologic effects and permeability rate of different suture materials in microsurgical anastomosis of the left uterine horn in Wistar rats. STUDY DESIGN: In this histologic study, 81 female Wistar rats were used. A transverse section and anastomosis were performed using three surgical techniques. The rats were divided into 15 experimental groups based on evaluation of three variations in microsurgical technique: (1) one-layer transmucosal anastomosis, (2) one-layer extramucosal anastomosis, and (3) two-layer transmucosal anastomosis. With each technique, five suture materials were used: 7/0 polyglycolic acid, 6/0 polyglactin, 7/0 polydioxanone, 7/0 silk and 7/0 polypropylene. RESULTS: The permeability rate was 86%. Polydioxanone persisted in 41% of cases at 90 days. Abnormal proliferation of mucosal epithelium was verified in 10% of cases. Polypropylene was responsible for the majority of foreign-body granulomas. Polydioxanone produced the smallest inflammatory reaction. CONCLUSION: The type of suture material did not influence the permeability rate. Use of nonabsorbable suture material altered the histologic structure the most and produced the greatest tissue reaction. The best results were obtained with polydioxanone. Monofilial sutures gave rise to fewer structural alterations than did multifilament sutures. PMID- 7473456 TI - Laser-assisted tubal anastomosis. AB - OBJECTIVE: To determine if laser-assisted anastomosis is superior to microsurgical anastomosis and laser welding for tubal reconstruction. STUDY DESIGN: Sixty uterine tubes from 30 rabbits were transected and then anastomosed with a microsurgical technique, laser welding and laser-assisted anastomosis using a microscope. The rabbits were mated one month postoperatively and examined for implantation sites and adhesion formation three to seven days postpartum. RESULTS: The amount of time required to perform laser-assisted anastomosis as well as laser welding was significantly shorter than for microsurgery. All the tubes withstood the distension pressure of pregnancy with the exception of 1 of the 20 laser-welded tubes, which had 30% of its circumference dehisced. There was no difference in the number of implantation sites per tube between the different groups. The anastomotic sites were well healed and were nonidentifiable except for holding stitches and microsurgical sutures. CONCLUSION: Laser-assisted anastomosis and laser welding took less time to perform than microsurgery. Laser assisted anastomosis resulted in excellent healing, as did microsurgery. With the protection of serum albumin, laser-assisted anastomosis did not cause any thermal damage, and the anastomotic sites could tolerate the distension pressure of pregnancy and parturition without problems. Laser welding without protection of serum albumin could cause thermal damage and dehiscence. The implantation and pregnancy rates were comparable The implantation and pregnancy rates were comparable with all three types of procedure. PMID- 7473457 TI - Discrimination against men in infertility treatment. AB - OBJECTIVE: To evaluate the scope of the medical effort invested in the study and treatment of male infertility, some of its sources and implications. STUDY DESIGN: The study reviewed the range of medical treatments of male and female infertility and their effectiveness. It also examined the funding policy in Ontario, Canada, as well as patients' responses to this policy. RESULTS: Male infertility therapy is relatively lacking and is not funded by the Ontario Health Insurance Plan. Neither professionals nor patients, however, challenge the unequal policy. CONCLUSION: The traditional association of infertility with the female partner prevails among professionals, policy makers and patients. It explains both the relative lack of therapy and the absence of public resentment. This traditional attitude hinders the prospects that infertile men will be assisted by biomedicine. PMID- 7473458 TI - Epidemiology of uterine leiomyomas. With an etiologic hypothesis. AB - OBJECTIVE: The few previous epidemiologic studies of uterine myomas have relied on clinical evaluation to select controls, but we previously showed that myomas may be present in more than 75% of such uteri. STUDY DESIGN: We therefore attempted to evaluate risk factors using age-matched controls whose uteri were serially sectioned to exclude the presence of myomas. RESULTS: The small study size precluded a meaningful evaluation of most parameters but tended to confirm the negative association of myomas with cigarette smoking (P = .07). CONCLUSION: Using monoclonal smooth muscle proliferation in human atherosclerotic plaques as a model, we suggest that excessive injury to and repair of the endometrial lining of the uterus may promote monoclonal expansion of smooth muscle cell populations in the uterine wall (i.e., myomas). This theory is largely compatible with the estrogen hypothesis, but fundamental principles of tumorigenesis and previous epidemiologic data on myomas suggest that nutritional factors should be scrutinized as possible initiators (DNA-damaging substances) in the pathogenesis of uterine myomas. PMID- 7473459 TI - Puerperal inversion of one horn of a bicornuate uterus. A case report. AB - BACKGROUND: Acute uterine inversion is a rare (1:2,500-25,000 deliveries) but potentially life-threatening obstetric complication. Uterine malformation can make it difficult to diagnose and treat this emergency. CASE: Following a normal delivery in a nullipara, laparotomy was required to establish the diagnosis and treat an inversion of one horn of a bicornuate uterus. CONCLUSION: Inversion of one horn of a bicornuate uterus presents a diagnostic dilemma. When physical examination reveals a palpable abdominal "fundus" and a mass protruding through the cervix in a patient who is experiencing uterine hemorrhage, the cause may be inversion of one horn of a bicornuate uterus. Laparotomy may be required for definitive diagnosis and treatment. PMID- 7473460 TI - Pasteurella multocida isolation from a tuboovarian abscess. A case report. AB - BACKGROUND: Human infection with Pasteurella multocida occurs most commonly following a nonhuman animal bite wound or cat scratch, with local skin and soft tissue infections the most frequent posttraumatic manifestations. Genitourinary tract pathology attributable to this organism occurs infrequently, with only one previous reported case of P multocida infection presenting in the form of a tuboovarian abscess. CASE: A 44-year-old woman with an acute abdomen underwent exploratory laparotomy, revealing a right-sided tuboovarian abscess. Following a total abdominal hysterectomy and bilateral salpingo-oophorectomy with a seven-day postoperative course of intravenous metronidazole, ampicillin and gentamicin, the patient was sent home on a regimen of cephradine, only to return with sepsis three days later. A course of intravenous aqueous penicillin sodium, metronidazole and cefotaxime was administered for the treatment of P multocida sepsis since this organism was identified in the intraoperative pelvic fluid culture 24 hours after the patient's initial discharge. After 13 days of the above regimen, the patient achieved full defervescence and was discharged on hospital day 15. CONCLUSION: In the setting of a tuboovarian abscess, the clinician should consider P multocida as a potential etiologic agent, especially in a patient with extensive exposure to nonhuman animals. In the treatment of an acute adnexal infection secondary to this organism, one should employ perioperative therapy with the appropriate antibiotics for a duration of at least 14 days. PMID- 7473461 TI - Cornual heterotopic pregnancy and cornual resection after in vitro fertilization/embryo transfer. A report of two cases. AB - BACKGROUND: Heterotopic pregnancy occurs in about 1% of pregnancies achieved with assisted reproductive technologies. The incidence of cornual pregnancy is approximately 1% of all ectopics. CASES: Two patients became pregnant after treatment with IVF-ET. Both presented during the first trimester with evidence of an acute abdomen and ultrasonographic suspicion for cornual heterotopic pregnancies. The diagnoses were confirmed by laparoscopy, and treatment was undertaken with laparotomy with cornual resection. Both delivered viable infants by cesarean section; the first patient delivered a twin gestation at 28 weeks and the second a singleton pregnancy at 37 weeks. All infants were doing well two months after delivery. CONCLUSION: Abnormal pregnancies can be detected after IVF ET with careful follow-up, transvaginal ultrasound studies, serial hCG measurements and pelvic examinations. Cornual heterotopic pregnancies can be successfully treated with cornual resection, especially in cases presenting during the second trimester or when there is clinical evidence of an acute abdomen. All deliveries should be performed by cesarean section at term prior to labor or when tocolysis for premature labor has failed. PMID- 7473462 TI - Classification criteria for childhood arthritic diseases. PMID- 7473463 TI - Neurogenic influences in arthritis: potential modification by capsaicin. PMID- 7473464 TI - HLA-DRB1 genotyping in patients with rheumatoid arthritis in Taiwan. AB - OBJECTIVE: To investigate the relation of HLA-DR alleles to the development of rheumatoid arthritis (RA) and the correlations between HLA-DR alleles and clinical manifestations of patients with RA in Taiwan. METHODS: The HLA-DRB1 alleles were studied in 144 patients with RA and 154 healthy controls using polymerase chain reaction/sequence specific oligonucleotide probe methods. The subtypes of HLA-DR4 were detected by cloning sequencing. RESULTS: The prevalence of HLA-DR4 was significantly higher in patients with RA (46.53%) than in healthy controls (25.97%). HLA-DRB1*0405 was more common in patients than in controls (83.58 vs 50%), while the prevalence of DRB1*0403 in patients (4.48%) was significantly lower than in controls (27.5%). There was no significant difference in frequencies of HLA-DR alleles between DRB1*0405 negative patients and DRB1*0.05 negative controls. We also noted a positive correlation in patients between HLA-DR4 and bone erosion, but not HLA-DR4 and age of onset, seropositivity, or extraarticular involvement. There was no significant difference in seropositivity, extraarticular involvement, and bone erosion between homozygous and heterozygous DR4 patients. CONCLUSION: HLA-DRB1*0405 is related to the development of RA in Taiwan, while DRB1*0403 was a negative risk factor. HLA-DR4 was associated with bone erosion in patients. PMID- 7473465 TI - Sex hormone status and bone mineral density in men with rheumatoid arthritis. AB - OBJECTIVE: Sex hormone status has been proposed as an important determinant of low bone mineral density (BMD) in both men and women. Our objective was to study the relationship between sex hormones and BMD in patients with rheumatoid arthritis (RA), and how steroid therapy affects both. METHODS: We studied 99 men with RA to assess their BMD and sex hormone status. A comparative group of 68 age paired men was used. We made comparative tests, linear correlations, and multiple regression analysis. RESULTS: We found significant reductions in lumbar BMD (p = 0.0005), femoral BMD (p < 0.0005), salivary testosterone (p = 0.01), androstenedione (p = 0.007), and dehydroepiandrosterone sulfate (DHEAS) (p = 0.03) in patients with RA. In contrast, serum testosterone concentrations were normal. Salivary testosterone showed correlation with femoral BMD (r = 0.36; p < 0.001). By multiple regression analysis, weight, serum testosterone concentrations, and the cumulative dose of corticosteroids were significant predictors of lumbar BMD (r = 0.41; p = 0.001). Weight, age, androstenedione concentrations, and the cumulative dose of corticosteroids were the significant predictors of femoral BMD (r = 0.79; p < 0.0005). CONCLUSION: We confirm there is reduced BMD in men with RA; corticosteroids contribute, but are not the only factor in the pathogenesis of low BMD in patients with RA; we found decreased levels of androstenedione, DHEAS, and salivary testosterone in men with RA; and salivary testosterone, as with free testosterone, is correlated with BMD in patients with RA, with lower levels contributing to low BMD. PMID- 7473466 TI - Technetium-99m labelled polyclonal human IgG scintigraphy before and 26 weeks after initiation of parenteral gold treatment in patients with rheumatoid arthritis. AB - OBJECTIVE: To determine effectiveness of technetium-99m labelled polyclonal human immunoglobulin G (99mTc-IgG) scintigraphy to monitor variation in arthritis activity in patients with rheumatoid arthritis (RA). METHODS: The results of semiquantitative 99mTc-IgG scintigraphy were compared with those of examination before and 26 weeks after initiation of parenteral gold treatment in 19 patients with RA. RESULTS: Clinical and laboratory variables of arthritis activity as well as the scores of 99mTc-IgG scintigraphy were significantly lower after gold treatment compared to the scores before treatment. However, the difference between the mean scores of 99mTc-IgG scintigraphy before and after treatment was statistically significant for more joints than such difference in scores for joint pain and joint swelling. CONCLUSION: 99mTc-IgG scintigraphy is able to reflect variations in arthritis activity in patients with RA. PMID- 7473467 TI - Is it possible to predict the first year extent of pain and disability for patients with rheumatoid arthritis? AB - OBJECTIVE: To investigate prediction of the extent of suffering during the first year of rheumatoid arthritis (RA) with patient characteristics found to be capable of predicting short term or longterm endpoint disability. METHODS: Ninety five patients with recently diagnosed RA, participating in a prospective clinical trial, were seen every 3 mo during 1 year. The 12 month and cumulative values of disability (Dutch version of the Health Assessment Questionnaire) and pain (visual analog scale) were related to demographic, clinical, laboratory, and psychological mood variables in correlation and regression analyses. RESULTS: Baseline values of disability and pain were related most strongly to 12 mo and cumulative values of disability; baseline pain was virtually the only important predictor of 12 mo and cumulative pain. Other baseline characteristics had virtually no additional predictive power. CONCLUSION: The short term disease course of RA in terms of disability and pain is most strongly related to the baseline values of these variables, and cannot reliably be predicted with frequently recognized longterm prognostic factors, such as rheumatoid factor status or sex. PMID- 7473468 TI - Trends in hospitalizations for gastrointestinal bleeding among patients with rheumatoid arthritis in Rochester, Minnesota, 1950-1991. AB - OBJECTIVE: Estimates of the incidence of significant gastrointestinal (GI) bleeding among patients with rheumatoid arthritis (RA) vary and population based estimates of this event have been unavailable. We examine the incidence of this event in a well defined cohort of patients with RA. METHODS: The study cohort consisted of patients with RA incident in Rochester, MN, between 1950 and 1974 and reported by Linos, et al (Am J Epidemiol 1980;111:87-98). These patients were followed for up to 40.5 yrs, and retrospective analysis of hospitalization for GI bleeding was performed. RESULTS: A total of 58 patients with RA were hospitalized for GI bleeding, including 46 patients who had a first episode of bleeding after the diagnosis of RA was made, for a first episode incidence rate of 0.52% per person-year of followup. These 46 patients experienced 72 hospitalizations for GI bleeding. Patients diagnosed with RA after 1963 and patients older than 53 yrs of age at the time of diagnosis of RA had a higher incidence of hospitalization for GI bleeding. Overall survival among all patients with RA was slightly less than expected in the general population, while survival in patients hospitalized for GI bleeding was significantly lower than that of nonhospitalized patients. In a nested case-control study, nonsteroidal antiinflammatory drug use was found to be a significant risk factor for hospitalization for GI bleeding (odds ratio 3.25:1 compared with nonusers). CONCLUSION: Estimates of the overall incidence of hospitalization for GI bleeding in this population based cohort of patients with RA may be somewhat lower than some reported for referral populations, but an upward trend in recent years is noted. PMID- 7473469 TI - Risk of malignancy in women with systemic lupus erythematosus. AB - OBJECTIVE: To estimate the risk of malignancy in a cohort of patients with systemic lupus erythematosus (SLE) from a university medical center. METHODS: The cohort consisted of patients with lupus who were residents of Allegheny County and who were seen at the University of Pittsburgh, 1981-91. Cases of cancer were identified by a postal survey and review of medical records. The expected number of malignancies in the lupus cohort were estimated from age, sex, and race adjusted census and cancer incidence data from Allegheny County. Standardized incidence ratios and 95% confidence intervals for all cancers were calculated. RESULTS: Six (2.7%) malignancies were observed in the 219 lupus patients from Allegheny County during the observation interval (1981-91) and after the first visit at the University of Pittsburgh. The expected number of malignancies was 4.42. The standardized incidence ratio for cancer in the lupus cohort from Allegheny County was 1.36 (95% confidence interval 0.50-2.96). The frequency of malignancy was not increased in the small number of patients who had received immunosuppressive drugs before the diagnosis of cancer. Non-Hodgkin's lymphoma occurred in one patient, who also had Sjogren's syndrome, and in one patient before entry into this study who was treated with FK-506 following renal transplant. No bladder cancers were observed during the study interval. CONCLUSION: The overall frequency of malignancy was not increased in this medical center cohort of patients with lupus during a mean followup interval of 5.2 yrs. Longterm followup and a multicenter effort is needed to refine risk estimates of cancer in patients with lupus. PMID- 7473470 TI - Reduced muscle uptake of oxygen during exercise in patients with systemic lupus erythematosus. AB - OBJECTIVE: To assess the factors limiting aerobic exercise capacity in patients with systemic lupus erythematosus (SLE). METHODS: The anaerobic threshold (AT) and O2 pulse, i.e., VO2/heart rate (HR), were measured in 21 patients with SLE without cardiopulmonary complications by the analysis of expired gas during incremental work load in exercise testing. The relationships between work rate (WR) and VO2, HR and VO2/weight (Wt), and lupus activity index (LAI) and AT were analyzed. RESULTS: The AT of the patients with SLE was significantly lower than for a control group. Little increase in O2 pulse and very low delta VO2/delta WR and delta VO2/Wt@delta HR were found during work load in some patients with SLE, especially those with a low AT and high LAI. Patients with SLE with a high LAI tended to show a low AT. CONCLUSION: The low aerobic exercise capacity of patients with SLE appeared to be mainly due to a small increase in O2 pulse. This may have resulted from impaired oxygen diffusion in the inflamed peripheral muscles in patients with active SLE. Low AT may explain in part why patients with SLE become easily fatigued. PMID- 7473471 TI - Interleukin-2 receptor expression in salivary glands of patients with Sjogren's syndrome. AB - OBJECTIVE: To study interleukin-2 receptor (IL-2R) expression in lymphocytic and epithelial cells in salivary glands of patients with Sjogren's syndrome (SS). METHODS: Minor salivary glands of 29 patients with primary SS, 28 with secondary SS, 19 with autoimmune diseases without SS, and 17 controls were studied using immunohistochemical techniques. Sections were analyzed with 3 different antibodies reacting with CD25 molecule. RESULTS: The presence of IL-2R (CD25) was observed in minor salivary gland lymphocytic infiltrate in 26 (89.6%) primary SS, 24 (85.7%) secondary SS, and 16 (84%) autoimmune diseases without SS. CD25 antibody was expressed in epithelial cells in 12 (41%) patients with primary SS and 11 (39%) with secondary SS, but was absent in the group of autoimmune diseases without SS and the control group. CONCLUSION: The lymphocytic infiltrate of minor salivary gland expressed IL-2R in patients with primary SS, secondary SS and autoimmune diseases without SS. IL-2R expression in epithelial cells was only present in patients with primary or secondary SS. PMID- 7473472 TI - Deflazacort versus methylprednisolone in polymyalgia rheumatica: clinical equivalence and relative antiinflammatory potency of different treatment regimens. AB - OBJECTIVE: To assess the clinical efficacy and equivalence of a daily vs alternate day regimen, and the potency ratio between 2 glucocorticoids, deflazacort and 6-methylprednisolone. METHODS: Thirty-one patients with recent onset polymyalgia rheumatica (PMR) were randomly assigned to deflazacort (n = 16) or 6-methylprednisolone (n = 15), according to a 2 period (duration of each period = 6 weeks), crossover, open design for comparing 2 dose regimens (daily vs alternate day), and according to a between-patients, double blind design for comparing the therapeutic effects of the 2 glucocorticoids (deflazacort vs 6 methylprednisolone). The patients, either during alternate day or daily regimen, were treated with fixed oral doses for the first 2 weeks (assuming a potency ratio deflazacort/6-methylprednisolone of 1.5 mg/1.0 mg: deflazacort 24 mg or 6 methylprednisolone 16 mg, daily; deflazacort 48 mg or 6-methylprednisolone 32 mg, alternate day), and with titrated doses for the next 10 weeks. RESULTS: Two patients dropped out during the first 6 week period. The time course and extent of the improvement of disease activity indices (limb-girdle pain, morning stiffness, erythrocyte sedimentation rate, C-reactive protein, and plasma fibrinogen) were not statistically different under the 2 regimens. The satisfactory equivalent glucocorticoid response observed in 12 pairs of patients treated with deflazacort and 6-methylprednisolone progressed significantly from baseline to the end of the study, under daily or alternate day regimen, with no significant difference between the 2 glucocorticoids. Deflazacort proved less potent than 6-methylprednisolone (1.78 mg: 1.0 mg minimum effective daily dose; 1.68: 1.0, alternate day), but equally effective. CONCLUSION: The 2 dose regimens, 6-methylprednisolone and deflazacort showed no significant differences in terms of efficacy during the short term treatment of PMR. Deflazacort proved less potent than initially estimated. PMID- 7473473 TI - Prevalence of psoriatic arthritis in Italian psoriatic patients. AB - OBJECTIVE: To evaluate the prevalence of psoriatic arthritis (PsA) in Italian patients with psoriasis and to compare the Moll and Wright criteria, the European Spondylarthropathy Study Group (ESSG) criteria, and Amor criteria when applied to this patient population. METHODS: We examined 205 unselected patients with psoriasis. The diagnosis of PsA was based upon the clinical experience of a rheumatologist. After, we tested these classification criteria. HLA class I and II antigens were also analyzed. RESULTS: Thirty-six percent (75) of psoriatic patients were considered by a clinical expert to have PsA. Moll and Wright criteria identified 46 patients (22%) with PsA; 49 patients (24%) were identified by ESSG criteria and Amor criteria; 12 patients identified by Amor criteria but not by ESSG criteria presented enthesitis or dactylitis; 10 patients identified by ESSG but not by Amor criteria had peripheral synovitis. In patients with peripheral arthritis and psoriasis, the evaluation of NSAID response was critical to fulfilling Amor criteria. However, it was not easy to retrospectively evaluate NSAID response using these criteria. The sensitivity was low for each of the 3 classification criteria (from 61 to 65%), whereas the specificity was high (from 99 to 100%). CONCLUSION: Our study confirms a high prevalence of PsA among an unselected population of Italian patients with psoriasis. Our data reveal the inadequacy of the existing criteria for PsA, including the ESSG criteria and Amor criteria for the classification of spondyloarthropathy. PMID- 7473475 TI - Patterns of joint involvement in osteoarthritis of the hand: the Chingford Study. AB - OBJECTIVE: The hand is the major site of joint involvement in osteoarthritis (OA), but the clustering of this condition at different joint sites within the hand as part of an entity known as generalized OA remains contentious. We examine this issue in a population sample of 967 peri and postmenopausal women. METHODS: The pattern of radiographic involvement of hand joints was examined using the Kellgren and Lawrence system in a sample of women aged 45-64 years, selected from the general population in Chingford, East London. Log linear modelling techniques were used to examine clustering of OA in different joint sites and to examine whether this clustering remained significant after age adjustment. RESULTS: There was clear evidence of clustering in joint involvement: Thus 20 women had 4 or more affected joints compared with only 2 expected (chi 2 = 72.0, 4df, p < 0.001). The risk of multiple distal interphalangeal (DIP) joint involvement, after age adjustment, in a woman with a single affected DIP joint was significantly increased (OR 10.0, 95% CI 7.3-13.7) and was substantially greater than the corresponding risk for proximal interphalangeal joint (OR 3.1, 95% CI 1.4-6.8) involvement. Symmetry was the strongest determinant of pattern with an OR 38.8 (95% CI 14.5-103.5) for corresponding DIP joint involvement in the contralateral hand. CONCLUSION: These data provide clear evidence for a polyarticular subset of hand OA in women. There are 3 major determinants of the pattern of polyarticular involvement; symmetry, clustering by row and clustering by ray, in descending order of importance. PMID- 7473474 TI - D-lactic acid in synovial fluid. A rapid diagnostic test for bacterial synovitis. AB - OBJECTIVE: To analyze the usefulness of D-lactic acid levels in synovial fluid (SF) as a rapid test to support the early diagnosis of bacterial arthritis (BA). METHODS: A simple modification of the enzyme method used for measuring L-lactic acid was used to analyze levels of D-lactic acid in SF from 20 cases of BA. Results were compared with those from 99 noninfectious arthritis, which included 90 inflammatory SF samples. Total white blood cell count (WBC), percentage of polymorphonuclears (% PMN) and gram stains were also determined. RESULTS: D lactic acid levels were significantly higher in BA than in noninfectious arthritis. Using a cutoff value of 0.05 mM, 85% of the SF samples from BA had a positive test for D-lactic acid compared with 4% of the control group. The overall sensitivity of the assay was 85% with a specificity of 96%, showing a positive predictive value for BA of 81% and a negative predictive value of 97%. CONCLUSION: The data presented suggest that D-lactic acid is an accurate, easy test that can be carried out in any laboratory, to support the early diagnosis of BA. PMID- 7473477 TI - Interleukin-1 reversibly inhibits the synthesis of biglycan and decorin in intact articular cartilage in culture. AB - OBJECTIVE: To study the effect of interleukin-1 (IL-1) on the synthesis of proteoglycans biglycan (DSPG-I) and decorin (DSPG-II) in intact bovine articular cartilage. METHODS: Cartilage bearing sesamoid bones from the metacarpophalangeal joint were cultured with 10 ng/ml IL-1 for 2 days and labelled with [35S] sulfate. One sesamoid bone from each animal had been labelled ex vivo. The remaining 2 were cultured with IL-1 and allowed to recover in control medium before labelling. Control cultures were maintained in medium without IL-1 and labelled concurrently with the experimental series. The dermatan sulfate proteoglycans were purified from 4 M guanidinium chloride extracts of the cartilage by gel filtration on Sepharose CL-2B and CL-4B, on which they appeared as a single peak. Biglycan and decorin were separated by sodium dodecyl sulfide polyacrylamide gel electrophoresis in high salt. Individual lanes from the gel were cut in slices, which were dissolved and counted for radioactivity. RESULTS: Ex vivo, biglycan accounted for 4% and decorin for 2% of total incorporated sulfate. IL-1 reduced the synthesis of biglycan to 77% of the level of cultured controls and that of decorin to 73%. The synthesis of both proteoglycans returned to the control levels when the IL-1 was removed. IL-1 (10 ng/ml, 2 days) had no significant effect on total proteoglycan synthesis. CONCLUSION: The inhibition of synthesis of biglycan and decorin by IL-1 might be important in the pathophysiology of cartilage destruction in rheumatic diseases. PMID- 7473476 TI - Characterization of an additional articular cartilage vesicle fraction that generates calcium pyrophosphate dihydrate crystals in vitro. AB - OBJECTIVE: We previously identified a unique fraction of porcine articular cartilage vesicles, sedimentable at 8 x 10(6) g/min, which generate calcium pyrophosphate dihydrate crystals (CPPD) in vitro. We sought to identify and characterize other fractions of articular cartilage digest, sedimentable at lower g forces, which may also contain mineralizing vesicles. METHODS: Electron microscopy and alkaline phosphatase and nucleoside triphosphate pyrophosphohydrolase (NTPPPH) assays were used to analyze each fraction. Radiometric mineralization assays, Fourier transform infrared (FTIR) spectroscopy, and compensated polarized light microscopy were used to analyze crystals formed by these fractions. RESULTS: Vesicles of varying sizes identical to epiphyseal cartilage matrix vesicles were seen in all sedimentable fractions examined, but were the exclusive component of fractions sedimentable at 3 x 10(6) g/min, termed the heavy vesicle fraction (HVF), and at 8 x 10(6) g/min, now termed the light vesicle fraction (LVF). All vesicle containing fractions supported ATP dependent calcium pyrophosphate precipitation. The HVF and LVF precipitated 30 x more calcium than vesicle poor supernatant (p < 0.01) and 1.5-4 x more than cell-free unfractionated digest (p < 0.01). HVF differed from LVF in that it contained 3-4 x higher NTPPPH specific activity (p < 0.05). HVF resembled LVF in that both precipitated crystals consistent with CPPD by FTIR spectroscopy and compensated polarized light microscopy. CONCLUSION: These data expand our previous estimate of the total number of vesicles available for biologic mineralization and demonstrate heterogeneity of vesicle fractions. They support a key role for vesicles in CPPD crystal formation. PMID- 7473478 TI - Hydroxychloroquine in the treatment of erosive osteoarthritis. AB - OBJECTIVE: Hydroxychloroquine has been used successfully in the treatment of inflammatory arthritides such as rheumatoid arthritis and systemic lupus erythematosus, and less commonly in the seronegative spondyloarthropathies. Our objective was to determine the effectiveness of hydroxy-chloroquine in inflammatory or erosive osteoarthritis (OA). METHODS: We retrospectively reviewed clinical records of 8 patients prescribed hydroxychloroquine therapy whose symptoms of erosive OA were unresponsive to nonsteroidal antiinflammatory drugs (NSAID). RESULTS: Hydroxychloroquine was effective in 6 of the 8 patients treated; it was well tolerated by all patients. CONCLUSION: The use of hydroxychloroquine in patients with erosive OA unresponsive to NSAID appears promising. Prospective studies are needed to confirm our observations. PMID- 7473479 TI - Interobserver agreement on iliac crest pain syndrome in general practice. AB - OBJECTIVE: To investigate the degree of interobserver agreement on the iliac crest pain syndrome (ICPS) in patients with nonspecific low back pain in general practice. METHODS: Sixty-one patients with nonspecific low back pain were recruited by 11 general practitioners. All patients answered a questionnaire and underwent a standard examination. Each patient was examined by 2 independent observers. RESULTS: Kappa for ICPS was 0.57 (95% CI: 0.34-0.79). For the "localized tenderness" and "typical pain" criteria, kappa was 0.57 (95% CI: 0.40 0.73) and 0.66 (95% CI: 0.48-0.83), respectively. CONCLUSION: The presence of ICPS can be judged with good interobserver agreement. Clear definition of the diagnostic criteria and examining techniques, and multiple training sessions before and during the study are prerequisites. ICPS represents a group of patients with low back pain with the same symptoms and signs. A homogeneous group facilities studies on natural history, prognosis, and therapy. PMID- 7473481 TI - 31P NMR spectroscopy and electromyography during exercise and recovery in patients with fibromyalgia. AB - OBJECTIVE: To investigate whether patients with fibromyalgia (FM) have normal motor unit recruitment in relation to muscle metabolism during exhausting exercise and recovery, and whether the reduced voluntary muscle force normally seen is related to a smaller muscle size. METHODS: Female patients with FM and sedentary controls were examined using simultaneous 31P nuclear magnetic resonance spectroscopy (NMR) and surface electromyography (SEMG) during exhaustive static exercise of the anterior tibial muscle and during recovery. The maximum voluntary contraction force was estimated, and the maximum cross sectional muscle area was evaluated using 1H NMR imaging. The sedentary controls were matched to patients for sex, age and, as far as possible, daily physical activity levels. RESULTS: Patients with FM had reduced maximum voluntary contraction force in relation to the sedentary controls, despite having similar muscle size. In general the myoelectrical-metabolic relation during exercise and recovery was normal in patients with FM. CONCLUSION: The less extreme changes in motor unit recruitment and metabolism during exhaustive exercise indicated a lower exercise tolerance that could be connected with the lower physical activity levels. PMID- 7473480 TI - Patient utilities in fibromyalgia and the association with other outcome measures. AB - OBJECTIVE: To compare in patients with fibromyalgia (FM) utilities derived by rating scale and standard gamble methods; to gain insight into construct validity by relating utility values to other outcome measures; to assess the sensitivity to change of utilities. METHODS: A total of 73 patients with FM were randomized into one of 3 groups: low impact fitness training, biofeedback, or controls. At baseline and after 6 mo the Maastricht Utility Measurement Questionnaire was applied. By means of both the rating scale and standard gamble method patients were asked to value their own health status. Construct validity of patient utility measurements was evaluated by Spearman correlation and multiple regression of baseline values with pain, stiffness, patient's global assessment, Sickness Impact Profile (SIP), modified Health Assessment Questionnaire and Arthritis Impact Measurement Scale (AIMS). Sensitivity to change was assessed against changes in these outcomes. RESULTS: Rating scale utilities correlated significantly (p < 0.05) with patient's global assessment (rs = 0.53), pain (rs = -0.47), SIP (rs = -0.43), and with 9 of 11 dimensions of the AIMS (rs ranging from 0.23 to 0.62). Standard gamble utilities correlated significantly with mobility, pain, and arthritis impact of the AIMS scale (rs from 0.22 to 0.36) and with pain by visual analog scale (rs = -0.24) and patient's global assessment (rs = 0.32). Multiple regression analysis showed that patient's global assessment explained 41% (rating scale) and 10% (standard gamble) of total variance in baseline utilities. Also, 16% of the variance in change in rating scale utility values was explained by changes in patient's global assessment. In contrast, variance of changes in standard gamble utility values was not explained significantly by changes in other disease outcomes. CONCLUSION: Rating scale utilities correlated more strongly with disease outcome measures than standard gamble utilities. Also, construct validity for the rating scale was better than for the standard gamble. In FM, utility measurement is sensitive to the method chosen to elicit patient priorities. PMID- 7473482 TI - Pain coping mechanisms in fibromyalgia: relationship to pain and functional outcomes. AB - OBJECTIVE: To evaluate the factor structure of the Coping Strategies Questionnaire (CSQ) in patients with fibromyalgia (FM) and to compare the factors derived from this measure, along with the active and passive pain coping scales of the Pain Management Inventory (PMI) in predicting pain, depression, quality of well being (QWB), and pain behavior concurrently and over time. METHODS: One hundred twenty-two patients with FM were recruited from medical clinics, the community, and support groups. Eligible patients completed a battery of self report measures of pain and psychosocial functioning at baseline assessment before random assignment to a clinical trial. A subset of 69 patients who completed the clinical trial were readministered the same battery 3 mo later. Data were analyzed within the baseline period, and from the baseline period to posttreatment to evaluate the predictive effects of coping strategies on clinical outcomes. RESULTS: Principal components analysis of the CSQ revealed Coping Attempts (CA) and Pain Control and Rational Thinking (PCRT) factors, which have been found in other patient populations with chronic pain. Hierarchical multiple regression analyses revealed that high active coping and low PCRT contributed to higher concurrent pain, while low active coping and high passive coping were related to greater concurrent depression and pain behavior, respectively. Controlling for baseline scores on criterion measures, longitudinal multiple regression analyses demonstrated that high active coping and low PCRT scores contributed to greater pain, greater depression, and lower QWB at posttreatment, while low PCRT alone predicted greater pain behavior. CONCLUSION: The results show the import of the pain coping construct in FM and highlight the negative contribution of low perceived control over pain and high active coping to a range of pain outcomes. The findings on low perceived control converge with data on other chronic pain populations, while the role of active coping appears to be detrimental in FM, in contrast to its positive effects in patients with rheumatoid arthritis. PMID- 7473483 TI - A Grip Ability Test for use in rheumatology practice. AB - OBJECTIVE: To develop and evaluate a Grip Ability Test (GAT) for clinical evaluation of hand function in patients with rheumatoid arthritis (RA) by modifying a general test for hand function based entirely on activities of daily living, the Grip Function Test (GFT). METHODS: The GAT items were selected from those in the GFT that were sensitive to change in a hand training program of patients with RA. Based on a multivariate analysis, 3 items were chosen with an optimal representation of different grip types. RESULTS: The reliability was high: intraobserver test r = 0.985, p < 0.001; interobserver test r = 0.948, p < 0.001; internal consistency (Cronbach's alpha) 0.65. All items discriminated (p < 0.001) between patients with RA and healthy controls. The sensitivity to change in a hand training program in 24 patients with RA was significant for the total GAT (p < 0.001), as well as for the 3 items separately (p < 0.01, 0.01, and 0.05, respectively). CONCLUSION: The GAT for patients with RA was reliable, valid, and sensitive to change. It does not require sophisticated equipment and takes 5 min to perform. PMID- 7473484 TI - Proposal for the development of classification criteria for idiopathic arthritides of childhood. PMID- 7473485 TI - Influence of food on the bioavailability of oral methotrexate in children. AB - OBJECTIVE: To determine the bioavailability of oral methotrexate (MTX) in patients with juvenile rheumatoid arthritis in the fasting and fed states. METHODS: Each patient randomly received their usual weekly MTX dose either orally (po) after an overnight fast, po immediately after a breakfast of their choice, or intravenously (iv) on 3 consecutive weeks. Blood samples were taken at 0, 0.5, 1, 1.5, 2, 3, 4, and 6 h after po and 0, 0.08, 0.25, 0.5, 1, 1.5, 2, 3, 4, and 6 h after iv administration. RESULTS: Fourteen patients (10 female) aged 2.8 to 15.1 yrs completed the study; the results of 13 patients were evaluable. The mean elimination rate constant was 0.27 +/- 0.065, 0.26 +/- 0.067, and 0.25 +/- 0.11 h 1 after po fasting, po fed, and iv administration, respectively. The total area under the serum concentration vs time curve was 1.87 +/- 0.83, 1.50 +/- 0.51, and 1.85 +/- 0.80 mumol/l.h after po fasting, po fed, and iv administration, respectively. The maximum serum MTX concentration (Cmax) was 0.65 +/- 0.33 and 0.39 +/- 0.18 mumol/l after po fasting and po fed administration, respectively (p = 0.0022). The time to Cmax was 0.94 +/- 0.40 and 1.32 +/- 0.68 h after po fasting and po fed administration, respectively (p = 0.1464). The bioavailability of oral MTX while fasting was 1.1 +/- 0.51, while that after a meal was 0.88 +/- 0.35 (p = 0.0211). CONCLUSION: These data indicate greater oral bioavailability of MTX in the fasting state. We recommend that children receive MTX on an empty stomach. PMID- 7473486 TI - Frequency of relapse after discontinuation of methotrexate therapy for clinical remission in juvenile rheumatoid arthritis. AB - OBJECTIVE: To investigate the outcome of children with juvenile rheumatoid arthritis (JRA) who discontinued taking methotrexate (MTX) therapy after the achievement of clinical remission. METHODS: We conducted a retrospective review of the clinical course of all consecutive patients with JRA treated with MTX at our department. RESULTS: Seventeen of the 30 patients who responded to treatment entered clinical remission 6 to 30 mo after beginning MTX therapy. All patients were discontinued from MTX within 2 to 5 mo. Five patients relapsed within 9 mo, whereas 12 patients had sustained remission for 12 mo or longer. Four of 5 children with extended pauciarticular JRA relapsed early after MTX discontinuation. These patients were less responsive to a further cycle of MTX, and 3 showed progression of radiographic joint lesions. CONCLUSION: These findings suggest that MTX should be continued in patients with extended pauciarticular JRA even after the achievement of clinical remission. PMID- 7473487 TI - Elevated serum prolactin levels in children with juvenile rheumatoid arthritis and antinuclear antibody seropositivity. AB - OBJECTIVE: To determine if antinuclear antibody seropositivity in girls with juvenile rheumatoid arthritis (JRA) is associated with elevated serum levels of the anterior pituitary hormone prolactin. METHODS: Nineteen premenarchal girls meeting ACR classification criteria for JRA were evaluated for disease activity, antinuclear antibodies (ANA) seropositivity, and serum concentrations of prolactin, estrogen, and thyroid stimulating hormone. RESULTS: The mean serum prolactin concentration of ANA seropositive patients with JRA (10.9 +/- 1.9 micrograms/l) was significantly higher than that for ANA negative patients (5.7 +/- 1.0 micrograms/l; p = 0.043) and an age matched control group (5.8 +/- 1.3 micrograms/l; p = 0.048). CONCLUSION: Children with ANA seropositive JRA have elevated serum levels of the immunostimulatory hormone prolactin. PMID- 7473488 TI - Elevated circulating interleukin-7 levels in patients with systemic juvenile rheumatoid arthritis. AB - OBJECTIVE: Since interleukin-7 (IL-7) has been shown to induce monocyte production of IL-1, IL-6, IL-8, and tumor necrosis factor-alpha, we investigated plasma and synovial fluid levels of IL-7 in patients with juvenile rheumatoid arthritis (JRA). METHODS: IL-7 levels were measured using an ELISA in 60 patients with JRA and in 25 patients with other rheumatic disorders (ORD). RESULTS: In patients with systemic JRA, but not in patients with polyarticular or pauciarticular JRA or with ORD, plasma IL-7 levels were significantly higher (p < 0.0001) than those of healthy controls. IL-7 was undetected in all synovial fluid tested. Among patients with systemic JRA, those with persistent systemic symptoms had IL-7 levels significantly higher (p < 0.01) than those of patients in which systemic symptoms subsided. CONCLUSION: Plasma IL-7 levels are elevated in patients with systemic JRA and are associated with the presence and severity of systemic symptoms. PMID- 7473489 TI - Catastrophic antiphospholipid syndrome with fatal acute course in rheumatoid arthritis. AB - A 34-year old woman, with a 3 yr history of severe seropositive rheumatoid arthritis (RA) with lupus anticoagulant and anticardiolipin antibodies, developed a massive anterior myocardial infarction and ischemia of the lower extremities, with disseminated intravascular coagulation resulting from extensive tissue damage. Seven days after admission, she died of severe heart failure complicated by ventricular fibrillation. To our knowledge, this is the first documented case of fatal acute antiphospholipid syndrome in RA. PMID- 7473491 TI - Progressive multifocal leukoencephalopathy complicating treatment for Wegener's granulomatosis. AB - We describe a man treated with cyclophosphamide and prednisone for Wegener's granulomatosis who developed biopsy proven progressive multifocal leukoencephalopathy (PML). Upon discontinuation of immunosuppression he made a dramatic recovery. As immunosuppressant use is increasing we draw attention to this potentially fatal, but reversible neurologic disease. PMID- 7473492 TI - Charles Bonnet syndrome in temporal arteritis. AB - An 87-year-old woman presented with Charles Bonnet syndrome--the occurrence of formed visual hallucinations in sane aged individuals. This was followed by headaches and unilateral visual loss, and the diagnosis of temporal arteritis (TA) was confirmed by biopsy. Steroid therapy resulted in disappearance of hallucinations, which recurred 7 mo later, responding to an increase in steroid dosage. Charles Bonnet syndrome may be an early sign of decreasing visual acuity in aged individuals; thus, diagnosis of TA or exacerbation of established TA should be considered in such patients. PMID- 7473490 TI - Primary antiphospholipid syndrome evolving into systemic lupus erythematosus. AB - Since 1983 we have followed a total of 165 patients with antiphospholipid syndrome (APS). During the median followup period of 78 mo (range 12-336 mo), 3 of 80 patients with primary APS subsequently developed features of systemic lupus erythematosus (SLE) or lupus-like disease. One patient developed lupus-like disease 4 yrs and the other 2 developed full blown SLE more than 10 yrs after initial presentation of primary APS. Tissue typing in patients who developed SLE showed HLA antigens A2, A3, B35, Bw6, Cw4, DR7, DRw53, and DQ2 (Case 2); and A1, A3, B7, B8, Bw6, Cw7, DR4, DR15, DR51, DRw53, and DQ1 (Case 3). We report clinical features and genetic associations of these 3 patients. PMID- 7473493 TI - The beneficial effect of the estrogen antagonist tamoxifen on experimental systemic lupus erythematosus. PMID- 7473494 TI - Sex ratio in ankylosing spondylitis. PMID- 7473495 TI - Septic arthritis due to Mycobacterium marinum. PMID- 7473496 TI - A new scoring system for the Bath Ankylosing Spondylitis Metrology Index (BASMI) PMID- 7473497 TI - The Department of Health Cardiovascular Review Group and public health policy: an alternative view. PMID- 7473498 TI - The feeding of patients in hospital: a review. AB - The feeding of patients in hospitals is a topical issue. This review considers the development of feeding practices in British hospitals and highlights disturbing aspects which may have a bearing on recovery. The issues highlighted may also exacerbate the problem of hospital-induced malnutrition. There is little evidence that the provision of food per se is a cause for concern. It is rather the many other factors which may contribute to the patients consuming an in adequate diet. PMID- 7473499 TI - Cigarette smoking and cardiovascular disease. PMID- 7473500 TI - The role of UK tobacco advertising: no effect on the nation's health? PMID- 7473501 TI - Rheumatic heart disease in India. AB - A cross-sectional study was carried out in rural areas of Jawan Block, District Aligarh, Uttar Pradesh, India, covering a total population of 3760 drawn from 11 villages. The total number of confirmed cases of rheumatic heart disease was 24 with a prevalence rate of 6.4 per 1000 of the general rural population. The prevalence of rheumatic heart disease increased with age until the age of 25 years. Females were more prone to rheumatic heart disease compared to males. Socio-economic class had a direct impact on the occurrence of rheumatic heart disease. PMID- 7473502 TI - Surveillance of tuberculosis caused by Mycobacterium bovis in Slovakia. AB - The elimination of tuberculosis (TB) among cattle was claimed in Czecho-Slovakia in the middle of the sixties. Experiences from some countries which eliminated TB among cattle earlier than Czecho-Slovakia indicated a sporadic occurrence of TB caused by M. bovis. A long-term investigation of the occurrence of this 'diminishing zoonosis' has been carried out by a special group of experts (epidemiologists, clinicians, bacteriologists, epizootiologists and veterinary surgeons) on the whole territory of Slovakia. During the period of years 1972 1990 there were discovered 68 new cases of TB in humans caused by M. bovis. This paper analyses the results of the epidemiological investigation of this group of patients from several aspects: eg age, sex, occupation and geographical distribution. PMID- 7473503 TI - Mothers' attitudes to children's chest infections in India. AB - Acute Respiratory Infections (ARIs) account for a very high morbidity and mortality amongst children in the developing countries. A knowledge, attitude and practice study in relation to the literacy status of mothers whose children suffered from ARI was conducted. A sample of 140 mothers who had 265 children were selected for the study. The majority of literate mothers (75%) had complete knowledge regarding management of ARI. Literacy alone was not the only factor responsible for developing a positive attitude and adopting correct practices during ARI. Mass media and health personnel played an equally important role. Most women (89.3%) had obtained their knowledge regarding ARI through media and paramedical staff. Most mothers (96.4%) were concerned about the health of their children when they suffered from episodes of ARI and the majority of them (87.2%) were worried because they felt that their children or contacts could be adversely affected. Nearly 72% of mothers took early action during an episode of ARI. The majority of the medical practitioners practised non-allopathic medicine but all of them were prescribing allopathic drugs. Most of the mothers (70%) had no problem in taking their children to the desired health centers when needed. PMID- 7473504 TI - Grass-root health education strategies in Malawi. PMID- 7473505 TI - The physiological threat of compulsive syndromes. AB - Compulsive syndromes, especially obsessive-compulsive disorder, are much more common than previously believed. The behaviours that are a part of these syndromes can lead to physiologically threatening conditions whose medical consequences range from minor to fatal. It is crucial that not only mental health professionals, but all health professionals better understand the nature and consequences of compulsive disorders and their importance to general health. PMID- 7473506 TI - Care at home and quality of life: an overview of the issues. PMID- 7473507 TI - Independent sector home care: current issues and uncertainties. PMID- 7473508 TI - The great train crash approaches. PMID- 7473509 TI - AIDS' nomenclature's spelling or style. PMID- 7473510 TI - Vitamin C, smoking and SIDS. PMID- 7473511 TI - Certificate in basic oral health promotion. PMID- 7473512 TI - Proceedings of a symposium on cardiovascular research: Past triumphs and future prospects. Glasgow, Scotland, 7-8 October 1994. A festschrift for AF Lever, JJ Brown and JIS Robertson. PMID- 7473513 TI - Evolution of blockade of the renin-angiotensin system. PMID- 7473514 TI - Evidence for cardiovascular effects of prorenin. AB - This report investigates whether there is evidence that prorenin has cardiovascular effects. The report concludes that prorenin may indeed have cardiovascular effects, causing regional vasodilation by counteracting the vasoconstrictor effect of renin, and thereby maintaining blood flow to vital organs. This view is based on several observations. In direct contrast to renin, high levels of prorenin are not associated with vasoconstriction. (1) Prorenin is expressed almost exclusively in tissues with extraordinarily high levels of blood flow: pregnant uterus, placenta, ovaries, kidneys, eyes. (2) In the ovaries, the higher the prorenin level the higher the level of steroid biosynthesis, consistent with a increased tissue perfusion or metabolism. (3) Blood pressure gradually falls when prorenin is infused into monkeys, even when the source of prorenin is contaminated with renin which should increase blood pressure. (4) Prorenin levels positively correlate with renal blood flow under several experimental conditions. (5) Salt sensitivity of blood pressure, a renal vasoconstricted state, is associated with subnormal prorenin levels. (6) Diabetes mellitus and pregnancy, two vasodilated states, are associated with high plasma prorenin levels. (7) Prorenin binds to a membrane receptor that also recognizes renin. Thus, prorenin could vasodilate by competing with the specific uptake of renin and thereby reduce the level of regional vasoconstriction. Altogether, the cardiovascular haemodynamic setting associated with high levels of prorenin is the opposite of that associated with high levels of renin and is consistent with a vasodilator effect in the kidneys and reproductive organs and perhaps elsewhere. PMID- 7473515 TI - Renin-angiotensin-aldosterone system for blood pressure and electrolyte homeostasis and its involvement in hypertension, in congestive heart failure and in associated cardiovascular damage (myocardial infarction and stroke). AB - The renin-angiotensin-aldosterone hormonal servo-control system plays a major role in defending normotension and sodium and potassium balance. Derangements of the system involving either excess renin-angiotensin vasoconstriction or too much sodium-volume retention for the plasma-renin level can be implicated in sustaining the hypertension of most human hypertensive disorders. When plasma renin levels are inappropriately or inordinately increased, ischemic vascular injury to the heart, brain and kidneys becomes increasingly evident. In this setting, pharmacologic suppression or blockade of plasma renin activity protects from further cardiovascular damage in experimental models, in human hypertensive disorders and in human congestive heart failure. Still other studies suggest that excessive plasma prorenin levels may be involved in the vasodilator hyperperfusion vascular injury of diabetes mellitus. Recent findings provide considerable promise for pharmacologic strategies that suppress or block plasma renin activity in patients with hypertension or with congestive heart failure for achieving long-term protection from the major cardiovascular sequellae that shorten useful life. Future research to define better the factors that control renal renin gene expression and renal renin secretion could lead to even better treatment and prevention strategies. PMID- 7473516 TI - Mildly raised corticosterone excretion rates in patients with essential hypertension. AB - In a large group of patients with untreated hypertension, excretion rates of tetrahydrocorticosterone (THB), allotetrahydrocorticosterone (alloTHB) and tetrahydro-11-dehydrocorticosterone (THA) were all significantly higher than in a group of matched normotensive controls. Using the sum of these metabolites as an index of corticosterone secretion rate suggests that this variable is also higher. The increase was small (all excretion rates remained within the normal range) and corticosterone has low glucocorticoid and mineralocorticoid potency calling the clinical significance of this finding into question. However, it is possible that in combination with mildly deficient 11 beta-hydroxysteroid dehydrogenase activity such increases in corticosterone levels might affect blood pressure. PMID- 7473517 TI - Experimental studies on cortisol-induced hypertension in humans. AB - Studies in humans have shown that cortisol administration (200 mg/day) increases cardiac output, renal vascular resistance, glomerular filtration rate, plasma volume, extracellular fluid volume, exchangeable sodium, plasma glucose, insulin, renin substrate and atrial natriuretic peptide concentrations as well as urinary kallikrein excretion. Cortisol treatment decreases renin and angiotensin II concentrations while catecholamines and vasopressin are decreased or unchanged. We have clear evidence from a number of studies that cortisol-induced hypertension is modulated by, but not dependent on, exogenous sodium. The increase in cardiac output normally seen with cortisol administration is not essential for the blood pressure rise. The role of the increase in renal vascular resistance in the genesis of the hypertension is unclear. Studies using measurements of noradrenaline spillover and assessment of reflex function have not shown any increase in sympathetic nervous system activity but changes in vascular responsiveness, particularly to phenylnephrine and noradrenaline are marked. Cortisol is known to have a variety of effects on brain, heart, kidneys, blood vessels and body fluid volumes. To what extent the observed changes are epiphenomena, amplifiers or modulators, or are causal is unclear. Cortisol hypertension may reflect a complex interplay of these factors varying with the steroid concentrations achieved, underlying genetic factors and the particular experimental circumstances. PMID- 7473518 TI - Hypertension trial results: consensus and conflicts. AB - In severe and accelerated hypertension the benefits of treatment are clearcut. In patients < 60 years of age with mild hypertension the main benefit is reduction of stroke by about 40%. The death rate from stroke is declining in many affluent countries for reasons which can only be partly explained by mass treatment of hypertension. In the MRC trial in patients < 60 years old it took 2500 patient/years of treatment to save one stroke. If the number of strokes is declining for other reasons the number of patient/years to save one stroke may be increasing. In older patients the absolute benefit is greater because they suffer more stroke events and because treatment also reduces coronary events. PMID- 7473519 TI - Adjustment of the apparent benefits of treatment on stroke risk in the MRC mild hypertension trial using data from the placebo-treated group. AB - Clinical trials show that drug treatment significantly decreases stroke risk in hypertension. The benefit as measured in clinical trials may be affected by changes of blood pressure at entry and by departure from randomised treatment, but the magnitude of such effects is disputed. We have assessed benefit from reduction of stroke using data from the MRC Trial of mild to moderate hypertension, taking these factors into account, and have studied the likely effect of recent guidelines. The original analysis suggested that 850 patients needed treatment for 1 year to prevent one stroke. Under the more conservative of two assumptions made about the effect of treatment, this falls to 695 patients when allowance is made for reduction of stroke in placebo group patients withdrawn and actively treated, to 680 patients when allowance is made for the fall in blood pressure after entry, and to 556 patients with allowance for both. When benefit is assessed in patients whose entry diastolic blood pressure was > or = 100 mm Hg, 557 patients require treatment annually per stroke saved and this is decreased to 360 patients when allowance is made for withdrawal and active treatment of placebo group patients. These results suggest that benefit from reduction of stroke was underestimated in the MRC trial and that this is likely to be present in most trials. Changes to diagnostic criteria for hypertension in new management guidelines are likely to have significant effects on the number of patients treated per stroke prevented. PMID- 7473520 TI - Ambulatory blood pressure monitoring: from research to clinical practice. AB - Evidence from the medical literature is reviewed to indicate that ambulatory monitoring of blood pressure (ABPM) is a better predictor of target organ damage and clinical outcome in the hypertensive patient than clinic measurements of blood pressure (BP). A re-analysis of the documented BPs from the placebo limb of the Medical Research Council's treatment trial of mild hypertension is presented to indicate the difficulties inherent in the advice given by the various published guidelines on the diagnosis and management of hypertension. Finally, it is argued that because ABPM identifies a similar proportion of patients for treatment as a more prolonged follow-up, its use should be considered in the evaluation of all patients with mild hypertension as they can be categorised rapidly with less risk of being 'lost to follow-up'. PMID- 7473521 TI - Should cholesterol be measured in all hypertensives? AB - Guidelines for the management of hypertension, and those for hyperlipidaemia, advocate measurement of cholesterol in all hypertensive subjects. It is suggested that knowledge of serum cholesterol should influence the choice of anti hypertensive agent because thiazides and beta-blockers may influence lipids adversely. However, the changes in lipids associated with low-dose thiazides and beta-blockers are small, not sustained and do not appear to affect prognosis adversely. Some believe that knowledge of serum cholesterol may help target the treatment of mild hypertension more accurately by predicting an increased risk of vascular complications of hypertension. However, serum cholesterol does not predict the risk of cardiovascular complications in hypertensive women. It does predict coronary heart disease (but not stroke) in men, but coronary risk can be estimated satisfactorily without knowledge of serum cholesterol. Suggestions that cholesterol should be lowered in hypercholesterolaemic hypertensive patients to reduce the incidence of coronary heart disease are understandable. However, the diet recommended for this has no useful effect on serum cholesterol, and the benefit of lipid-lowering drugs for cholesterol reduction only exceeds the risk in patients at very high risk of coronary mortality, for example those who have had a myocardial infarction. Hypertension alone will not place individuals at high enough risk to warrant drug therapy for cholesterol reduction. We conclude that the current guidelines are incorrect, and that routine measurement of cholesterol in all hypertensive patients is not justified. PMID- 7473522 TI - Analysis of trough:peak ratio and the assessment of anti-hypertensive drug action. AB - The conventional blood pressure (BP) measurement is essentially a 'snapshot' at a single time point in any given 24 h period. In the clinical management of the hypertensive patient, however, it is assumed that this snapshot is representative of the BP throughout 24 h and indicative of the consistency of the control produced by anti-hypertensive drug treatment. However, anti-hypertensive drugs vary in the duration and consistency of their anti-hypertensive effect and a single measurement of BP can only be reliably indicative if the drug effect is known to be consistently maintained throughout the 24 h. Following the deliberations of the Food and Drug Administration in the USA, the calculation of a trough:peak ratio has been proposed as an index of the consistency of the anti hypertensive response and the suitability of an anti-hypertensive drug for its chosen dose and dose interval. In brief, to confirm that the magnitude of the BP is relatively consistent throughout 24 h it is recommended that the magnitude of the BP reduction at the end of the dose interval (at trough) should be at least 50% of the BP reduction measured at peak, namely a trough:peak of 50%. The trough:peak ratio provides a clinically relevant index for assessing whether an anti-hypertensive drug is likely to provide a consistent anti-hypertensive effect throughout the 24 h. PMID- 7473523 TI - Delivery of care for hypertension. AB - To overcome shortcomings in the delivery of care for hypertension various approaches have been developed including a system whereby care of hypertension is shared in a formal manner between general practitioners and hospital specialists. The feasibility, acceptability and cost-effectiveness of a computerised model of shared care were investigated in three matched groups of patients attending hypertension clinics in Glasgow. Glasgow Blood Pressure Clinic attenders considered suitable for shared care by their consultants were randomised to shared care (n = 277) or continued clinic follow-up; a further control group (n = 277) was identified from an independent nurse-practitioner clinic. After 2 years of follow-up, feasibility was estimated by the proportion of patients who had undergone an adequate review (blood pressure, serum creatinine and electrocardiograph); acceptability to general practitioners and patients was assessed by questionnaires; cost-effectiveness was calculated as the cost (to National Health Service and patient) per adequate review. The drop-out rate for shared care over 2 years was 3% compared with 14% for the outpatient clinic and 9% for the nurse-practitioner clinic. In year 2, rates of adequate reviews were 82%, 52% and 75%, respectively. Blood pressure control was similar in the groups. Of 297 general practitioners invited to participate, 85% agreed and 68% wished to continue participation after 2 years. About 50% of shared care patients preferred this method of follow-up compared with their earlier experience of clinic attendance. Shared care was more cost-effective than either conventional or nurse practitioner clinic follow-up, especially with respect to cost to the patient; costs per adequate review (pound sterling) were 28.96, 50.55 and 30.95, respectively.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7473524 TI - ACE inhibitors for the prevention and treatment of heart failure: why are they 'under-used'? AB - Currently there is concern that ACE inhibitors are not being utilised to their maximum potential for the prevention or treatment of heart failure. Underutilisation of ACE inhibitors has been attributed to a lack of appropriate medical education of physicians. However, an alternative explanation is that the trials have failed to answer important questions about how ACE inhibitors should be used in heart failure and that physician prescribing is appropriate to the current state of knowledge. The lack of evidence of benefit in patients with heart failure over the age of 75 years and of any substantial controlled trial in patients with apparent heart failure and well-preserved left ventricular systolic function, a common finding in elderly patients, may be common reasons why ACE inhibitors appear to be prescribed infrequently for heart failure. PMID- 7473525 TI - Mineralocorticoid receptor blockade in chronic heart failure. AB - The effects of ACE inhibitors on plasma aldosterone in chronic heart failure are poor, variable and unsustained. This is important to recognise because aldosterone has many harmful properties of its own. Aldosterone causes magnesium loss, potentiates catecholamines, induces ventricular arrhythmias, alters baroreflex function and stimulates myocardial fibrosis. Strategies to reduce these effects of aldosterone are being explored. PMID- 7473526 TI - Sick vessel syndrome: vascular changes in hypertension and atherosclerosis. AB - Hypertension and atherosclerosis are associated with structural and functional changes that may be collectively described as a 'sick vessel syndrome'. Structural changes in blood vessels (remodelling and hypertrophy) may be protective and adaptive. Functional changes in blood vessels include impairment of endothelium-dependent relaxation and impaired relaxation in response to activation of ATP-sensitive potassium channels. In general, vasorelaxation in response to direct activation of adenylate and guanylate cyclase is preserved in chronic hypertension and atherosclerosis. Vasoconstrictor responses to selected stimuli, such as serotonin, may be greatly potentiated. Impairment of endothelial function in combination with exaggeration of vasoconstrictor responses may predispose to vasospasm particularly during atherosclerosis. PMID- 7473527 TI - Effects of ACE inhibition on the heart. AB - Inhibition of the angiotensin converting enzyme (ACE) results in suppression of the formation of angiotensin II and delay of the degradation of bradykinin. Hence, the pharmacological effects of ACE inhibitors are attributable to both of these mechanisms. This is a brief review of the haemodynamic, neurohumoral and metabolic alterations following ACE inhibition, with an attempt to separate those attributable to the angiotensin-mediated actions and those attributable to bradykinin-mediated actions as they relate to prevention or attenuation of cardiac damage in hypertension, myocardial ischaemia and congestive heart failure. PMID- 7473528 TI - Endogenous endothelin generation maintains vascular tone in humans. AB - Endothelin-1 is a potent endothelium-derived vasoconstrictor and pressor peptide with uniquely sustained activity. We have examined the contribution of endogenously-generated endothelin-1 to the maintenance of basal vascular tone in healthy subjects. In these studies, on separate occasions, a combined inhibitor of endothelin converting enzyme (ECE) and neutral endopeptidase (NEP), phosphoramidon, a selective inhibitor of NEP, thiorphan, and a selective ETA receptor antagonist, BQ-123, were given via the brachial artery. Big endothelin 1, the precursor to endothelin-1, caused a slow onset dose-dependent forearm vasoconstriction, the magnitude of which was consistent with about 10% conversion to mature endothelin-1 in the forearm. Vasoconstriction to big endothelin-1 was abolished by co-infusion of phosphoramidon, whereas vasoconstriction to endothelin-1 was unaffected. Phosphoramidon caused progressive vasodilatation when infused alone, with blood flow increasing by 37% at 90 min (P = 0.02), whereas thiorphan caused vasoconstriction, consistent with NEP inhibition exerting its major effect on degradation of constrictor peptides, such as angiotensin and endothelin-1. Vasoconstriction to endothelin-1 was completely abolished by coinfusion of BQ-123, and BQ-123 alone produced progressive forearm vasodilatation, with blood flow increasing by 64% after 60 min (P = 0.001). These results demonstrate that endogenous production of endothelin-1 acts to sustain vascular tone in humans and indicate that ECE inhibitors and endothelium receptor antagonists may have therapeutic potential as vasodilators. PMID- 7473529 TI - Intracellular effects of free radicals and reactive oxygen species in cardiac muscle. AB - Oxygen-derived free radicals (FRs) and other reactive oxygen species (ROS) have been implicated in the deleterious aspects of myocardial infarction, neutrophil infiltration and post-ischaemic reperfusion. We studied their actions on the main intracellular organelles of Ca-compartmentation and force production (the sarcoplasmic reticulum (SR) and myofilaments) in rat heart preparations by using two forms of chemical 'skinning'. We recorded Ca(2+)-activated isometric tension or, in saponin-treated trabeculae where SR function is maintained, either tension alone or tension and [Ca2+] transients evoked by caffeine. A single, brief application of xanthine/xanthine oxidase (generating superoxide; O2-) rapidly and irreversibly inhibits Ca(2+)-activated force with a dose- and time-dependent action. The kinetics of residual force production are slowed. Rigor induction (by ATP withdrawal) before and during exposure to .O2- prevents this action, suggesting the .O2(-)-sensitive site is occluded in rigor. Myofilament Ca sensitivity and SR function were unaffected by .O2- or physiologically relevant [H2O2] (< 10 microM). Briefly applying 10-50 microM hypochlorous acid (HOCl) increased Ca-sensitivity and resting tension, but reduced Ca-activated force, in a manner consistent with 'rigor-like' crossbridges being involved. HOCl also provoked spontaneous Ca-release but reduced net SR Ca-uptake. Electron microscopy reveals that the myofilament lattice suffers a characteristic disruption by HOCl but not by .O2-. We conclude that FRs and ROS associated with myocyte dysfunction, reperfusion and inflammation could contribute to post-ischaemic myocardial dysfunction. PMID- 7473530 TI - Vascular structure, smooth muscle cell phenotype and growth in hypertension. AB - The media of most blood vessels is occupied by vascular smooth muscle cells. In areas they may be replaced by glomus cells, pericytes or renin-secreting myoepithelioid cells. These cells may be regarded as phenotypic variants of vascular smooth muscle cells. In disease, ordinary vascular smooth muscle cells may also assume a spectrum of phenotypes ranging from contractile to largely secretory types. Therefore, the phenotype of smooth muscle cells that is involved in a disease process such as hypertension or one of its complications is likely to affect the structural outcome in blood vessels. PMID- 7473531 TI - Resistance vessel growth and remodelling: cause or consequence in cardiovascular disease. AB - Haemodynamic evidence indicates that in essential hypertension, minimum vascular resistance and vascular pressor response are increased. According to the original concept of Folkow, this is most easily explained in terms of a generalised narrowing of the vasculature and an increase in the ratio of the vessel wall (more specifically, tunica media) thickness to lumen diameter. These predictions have been confirmed, both by histological and in vitro studies of resistance vessels. Moreover, the evidence indicates that these changes are associated mainly with remodelling (rearrangement of the same amount of material) of the vessels rather than growth. However, although the alteration in small artery structure is usually appropriate to the actual blood pressure (BP), the structure is not only a secondary adaptation but is also dependent on other factors, including neurohumoral and genetic factors. Furthermore, although the resistance vessel structure clearly plays an important role in determining BP, it does not appear to be a prime determinant of BP. It thus appears that more emphasis should be placed on Folkow's original idea that resistance vessel structure, like neurohumoral drive, is a mediator of the mechanisms which determine BP. Therefore, although evidence is available that the structure of certain specific parts of the vasculature (namely the renal afferent arterioles) could play a dominant function in the development of hypertension, it is suggested that resistance vessel structure should not in general be considered to have this role. PMID- 7473532 TI - Does the rapidity with which hypertension reverses on unclipping a clipped kidney and on stopping long-term infusion of angiotensin II make unlikely an important role for vascular hypertrophy in maintaining the hypertension seen before reversal? PMID- 7473533 TI - Relation between renin and prorenin in plasma from hypertensive patients and normal people: evidence for different renin:prorenin ratios. AB - Renin and prorenin concentrations in plasma correlate closely, but the proportion of active renin to prorenin shows some variation in different forms of hypertension. The proportion of active renin to prorenin is higher in normal renin essential hypertensives and in patients with renal hypertension than in normal subjects, and it is lower in low renin essential hypertension and primary hyperaldosteronism. Plasma deficient in plasma prekallikrein shows a lower proportion of active renin than might be expected. Active renin and plasma angiotensin II concentration show a strong correlation while prorenin correlates weakly with plasma angiotensin II concentration. PMID- 7473534 TI - Vascular smooth muscle polyploidy in genetic hypertension: the role of angiotensin II. AB - Flow cytometry DNA analysis has been used to measure the percentage of aortic vascular smooth muscle cells in G2 + M phase of the cell cycle in mature stroke prone spontaneously hypertensive rats (SHRSP). The effects of three different pharmacological interventions on the cell cycle parameters have also been studied. Vascular smooth muscle cells isolated from SHRSP have significantly elevated G2 + M phase of the cell cycle compared with cells from the normotensive reference strain, Wistar-Kyoto (WKY). This observation reflects an increased tetraploid and octaploid cell populations in vivo. Treatment with a combination of hydralazine and hydrochlorothiazide had no effect on the percentage of cells in G2 + M phase of the cell cycle. Treatments with angiotensin converting enzyme inhibitor, perindopril or AT1 receptor antagonist, losartan, resulted in an equivalent blood pressure-lowering effect to that obtained with hydralazine/hydrochlorothiazide. In contrast to hydralazine/hydrochlorothiazide, these two treatments resulted in a highly significant regression of vascular smooth muscle polyploidy in the SHRSP. We hypothesise that angiotensin II plays an important role in cell cycle regulation in that, alone or in conjunction with one of the inhibitory proteins, it is able to stop the cell cycle progression after endoduplication but before the cytoplasmic division. Pharmacological interventions which remove an excess of angiotensin II may allow the cells to re enter the cell cycle thus resulting in the regression of vascular smooth muscle polyploidy and improved arterial compliance. PMID- 7473535 TI - SA gene and hypertension. AB - The SA gene was initially identified by differential hybridisation because of its higher expression in the kidney of the spontaneously hypertensive rat (SHR) compared with the normotensive Wistar-Kyoto rat. In subsequent studies, the allele of the SA gene from the SHR and several other genetically hypertensive strains was found to cosegregate with increased blood pressure in F2 progeny derived from crosses with normotensive rats. The increased expression of the SA gene in the kidney of the SHR occurs early, before the rapid rise in blood pressure in this model, is genotype-dependent and localised to the proximal tubule. Although the functions of its protein product remain to be elucidated, these findings raise the exciting possibilities that: (i) SA represents a major component of an important novel system regulating blood pressure, and (ii) it underlies a primary renal mechanism predisposing to hypertension. PMID- 7473536 TI - Dexamethasone-suppressible hyperaldosteronism: clinical, biochemical and genetic relations. AB - Clinical, biochemical and molecular data on five kindreds with dexamethasone suppressible hyperaldosteronism are reviewed. The clinical phenotype varies from severe, early onset hypertension to much milder blood pressure elevation; hypokalaemia is usually mild. The genetic basis of the syndrome reflects the presence of a chimaeric gene derived from an unequal crossover between CYP11B1 and CYP11B2, resulting in ACTH-sensitive aldosterone synthase activity. In five kindreds, at least three different mutations have been identified, suggesting that allelic predisposition might lead to increased geographical prevalence of the condition in Celtic populations. PMID- 7473537 TI - Salt intake: potential deleterious effects excluding blood pressure. AB - Increasing evidence from animal, epidemiological, intervention studies and treatment trials all clearly point to the important role that salt intake plays in determining blood pressure (BP). Of more current interest is whether salt may have other deleterious effects independent of its role in high BP. For instance, salt intake plays an important role in left ventricular hypertrophy independent of its effect on BP. Experimental evidence and some epidemiological evidence also suggest that salt intake may have an adverse effect on stroke mortality which may be independent of its effect on BP. In animal models of renal failure, dietary salt restriction has been found to slow the progression of the renal impairment, but no studies as yet have been reported in humans. Salt intake has also been associated with asthma, stomach and nasopharyngeal cancer. Increasing salt intake produces changes in the chemical composition of urine, particularly an increase in calcium excretion which will predispose to kidney stone formation and has also been shown to increase hydroxyproline excretion indicating increased bone resorption. It is likely that a high salt intake may be one of several factors aggravating osteoporosis. This may be particularly relevant in patients with essential hypertension who already have an increased urinary calcium excretion and may in the long term be at greater risk of osteoporosis. Restriction of salt intake reduces urinary calcium excretion and, perhaps, like thiazide diuretics may be beneficial in the long-term prevention of bone demineralisation. PMID- 7473538 TI - Salt and blood pressure revisited. AB - Modern attempts to link intake of salt with population blood pressure (BP) show striking similarity to early 19th century attitudes which attributed human disease and premature death to inappropriate diet. Under these circumstances there is a danger that conviction of the truth of a hypothesis may lead to selection of evidence. A pooled meta-analysis of rigorously conducted studies of the effect of salt restriction on BP has been carried out. This indicates small but real effects in hypertensive patients although the degree of heterogeneity cannot be assessed. The effect in normotensive subjects is minimal and throws doubt on the value of general reduction in salt intake in lowering population BP. PMID- 7473539 TI - Therapeutic approaches related to amyloid-beta peptide and Alzheimer's disease. PMID- 7473540 TI - Alkane-6,8-diol: inhibitor of tumor promotion in two-stage carcinogenesis in mouse skin. PMID- 7473541 TI - Phenoxypropylamines: a new series of squalene synthase inhibitors. PMID- 7473542 TI - Synthesis of novel 2,4-diaminopyrrolo-[2,3-d]pyrimidines with antioxidant, neuroprotective, and antiasthma activity. PMID- 7473543 TI - Cyclosporin analogs modified in the 3,7,8-positions: substituent effects on peptidylprolyl isomerase inhibition and immunosuppressive activity are nonadditive. AB - Four analogs of cyclosporin A (CsA) were synthesized to determine if the biological activities of CsA analogs generated by multiple amino acid replacements are predictable from the effects on biological activity of analogs with single residue changes. CsA analogs [Phe7]CsA (8a), [D-MeAla3,Phe7]CsA (8b), [D-Ser8,Phe7]CsA (8c), and [D-MeAla3,Phe7,D-Ser8]CsA (8d) were designed by modification of positions 3, 7, and 8, which are adjacent to one effector region of the cyclophilin-bound CsA complex. The syntheses of CsA analogs 8a-d were carried out by suitable modifications of the reported strategy. Each analog was characterized by NMR in deuterated chloroform and DMSO solutions, and their biological activities as inhibitors of cis-trans-peptidyl prolyl isomerase (PPIase), inhibitors of proliferation in BDF1 mouse spleen cells stimulated with concanavalin A (Con A), and inhibitors of IL-2 release stimulated with PMA/ionomycin by Jurkat cells were determined. Incorporation of the phenylalanine residue in position 7 diminished activities 5-8-fold. Substitution at position 3 decreased activity nearly 2-fold, and substitution at position 8 did not lower activities. However, when all three modifications (D-MeAla3,Phe7, and D-Ser8) were incorporated into one molecule, the resulting analog, 8d, was found to bind more tightly to cyclophilin than CsA (Ki = 3 +/- 1.5 vs 6 +/- 2 nM) and to produce the full immunosuppressive effect in the other assay systems. Our structure-activity results show that combinations of substitutions that individually lower PPIase or immunosuppressive activity produce fully active analogs when combined in a single compound. These results suggest that other, multimodified CsA derivatives may be discovered that possess excellent or improved immunosuppressive activities even though they contain a substitution that otherwise reduces immunosuppressive activity. PMID- 7473544 TI - Hydroxylated aromatic inhibitors of HIV-1 integrase. AB - Efficient replication of HIV-1 requires integration of a DNA copy of the viral genome into a chromosome of the host cell. Integration is catalyzed by the viral integrase, and we have previously reported that phenolic moieties in compounds such as flavones, caffeic acid phenethyl ester (CAPE, 2), and curcumin confer inhibitory activity against HIV-1 integrase. We now extend these findings by performing a comprehensive structure-activity relationship using CAPE analogues. Approximately 30 compounds have been prepared as HIV integrase inhibitors based on the structural lead provided by CAPE, which has previously been shown to exhibit an IC50 value of 7 microM in our integration assay. These analogues were designed to examine specific features of the parent CAPE structure which may be important for activity. Among the features examined for their effects on inhibitory potency were ring substitution, side chain length and composition, and phenyl ring conformational orientation. In an assay which measured the combined effect of two sequential steps, dinucleotide cleavage and strand transfer, several analogues have IC50 values for 3'-processing and strand transfer lower than those of CAPE. Inhibition of strand transfer was assayed using both blunt ended and "precleaved" DNA substrates. Disintegration using an integrase mutant lacking the N-terminal zinc finger and C-terminal DNA-binding domains was also inhibited by these analogues, suggesting that the binding site for these compounds resides in the central catalytic core. Several CAPE analogues were also tested for selective activity against transformed cells. Taken together, these results suggest that the development of novel antiviral agents for the treatment of acquired immune deficiency syndrome can be based upon inhibition of HIV-1 integrase. PMID- 7473545 TI - Effective inhibitors of hemagglutination by influenza virus synthesized from polymers having active ester groups. Insight into mechanism of inhibition. AB - Highly effective sialic acid-containing inhibitors of influenza virus X-31 were synthesized using poly[N-(acryoyloxy)succinimide] (pNAS), a polymer preactivated by incorporation of active ester groups. Polymers containing two and three different components were prepared by sequential reaction of pNAS with two and three amines, respectively. This preparation of co- and terpolymers was synthetically more efficient than methods involving copolymerization of different monomers and gave polymers that were more easily compared than those generated by copolymerization. Polymers in this study (prepared from a single batch of pNAS) had a constant degree of polymerization (DP approximately 2000) and probably had a distribution of components that was more random than analogous polymers prepared by copolymerization. Use of C-glycosides of sialic acid made it possible to investigate inhibition by different polymers at temperatures ranging from 4 to 36 degrees C without artifacts due to the hydrolytic action of neuraminidase. The inhibitors were, in general, more effective at 36 degrees C than at 4 degrees C. The hemagglutination (HAI) assay was used to measure the value of the inhibition constant KHAIi each polymer. The value of KHAIi for the two-component polymer containing 20% sialic acid on a polyacrylamide backbone at 4 degrees C was 4 nM (in terms of the sialic acid moieties present in solution) and was approximately 50-fold more effective than the best inhibitors previously described and 25-fold more effective than the best naturally occurring inhibitor. The most effective inhibitor synthesized in this work contained 10% benzyl amine and 20% sialic acid on a polyacrylamide backbone, and its value of KHAIi was 600 pM at 36 degrees C. Approximately 100 polymers that differed in one or two components were assayed to distinguish between two limiting mechanisms for inhibition of the interaction between the surfaces of virus and erythrocytes: high-affinity binding through polyvalency, and steric stabilization. The results suggest that both mechanisms play an important role. The system comprising polyvalent inhibitors of agglutination of erythrocytes by influenza provides a system that may be useful as a model for inhibitors of other pathogen-host interactions, a large number of which are themselves polyvalent. PMID- 7473546 TI - Esters of 3-pyridylacetic acid that combine potent inhibition of 17 alpha hydroxylase/C17,20-lyase (cytochrome P45017 alpha) with resistance to esterase hydrolysis. AB - Esters of 3- and 4-pyridylacetic acid have been prepared and tested for inhibitory activity toward the human testicular 17 alpha-hydroxylase/C17,20-lyase and human placental aromatase enzymes. The structural features required for optimal inhibition of the hydroxylase/lyase enzyme were a 3-pyridine ring, methyl substitution alpha to the carbonyl group, and a bulky alkoxycarbonyl substituent. The compounds with the greatest selectivity were isopinocampheyl 2-methyl-2-(3 pyridyl)propanoate, 9, 1-adamantyl 2-methyl-2-(3-pyridyl)propanoate, 12, and 2 methyl-2-adamantyl 2-methyl-2-(3-pyridyl)propanoate, 14, which, while inhibiting the aromatase activity with IC50 values of 30, 35, and 40 microM, respectively, exhibited IC50 values toward hydroxylase/lyase of between 13 and 90 nM. For comparison, ketoconazole gave an IC50 value of 15 microM against aromatase and values of 65 and 26 nM for inhibition of the hydroxylase and lyase activities, respectively. Some of the structural features required for enzyme inhibition also conferred resistance to esterase hydrolysis, in vitro using rat liver microsomes as a source of the esterase activity. Therefore these esters are lead compounds in the development of inhibitors of androgen biosynthesis for the treatment of hormone-dependent prostatic cancer. PMID- 7473547 TI - Piperazinylalkyl heterocycles as potential antipsychotic agents. AB - We recently reported on a series of pyrrole Mannich bases orally active in inhibiting the conditioned avoidance response (CAR) in rats. These compounds exhibit affinity for both D2 and 5-HT1A receptors, and some are noncataleptogenic. Such a profile suggests that they may be potential antipsychotic agents which lack the propensity for causing extrapyramidal side effects and tardive dyskinesias in humans. One of these compounds, 1-[[1-methyl-5 [[4-[2-(1-methylethoxy)phenyl]- 1-piperazinyl]methyl]-1H-pyrrol-2-yl]methyl]-2 piperidinone (RWJ 25730, 1), was chosen for further development but found to be unstable in dilute acid. In order to improve stability, we replaced the pyrrole methylene linkage to the piperazine ring with ethylene, employed ethylene and dicarbonyl as linkers between the lactam and the pyrrole ring, placed electron withdrawing groups on the pyrrole ring, and substituted acyclic amide for lactam. In addition, we replaced the pyrrole segment with other heterocycles including thiophene, furan, isoxazole, isoxazoline, and pyridine. Generally, replacement of the N-methylpyrrole segment with thiophene, furan, isoxazoline, or pyridine afforded compounds equipotent with 1 in CAR, which were more stable in dilute acid. In the case of side chain or lactam modifications, CAR activity was significantly decreased or abolished, with the exception of 6. For the most part, the modifications to 1 resulted in the decrease or loss of D2 receptor binding. However, within this series, 5-HT1A receptor binding was greatly increased, with thiophene 40 exhibiting an IC50 of 0.07 nM. The CAR activities of pyrroles 6 and 12, thiophene 40, furans 44-47, isoxazolines 49 and 50, and pyridine 54 coupled with their weak or nonexistent D2 binding and strong 5-HT1A binding suggest that they may be acting via a nondopaminergic mechanism or that dopaminergic active metabolites are responsible. Pyrrole 6 and furans 44 and 47 show promise as antipsychotic agents based on their CAR activity, receptor-binding profile, and solution stability. PMID- 7473548 TI - N-aryl-N'-benzylpiperazines as potential antipsychotic agents. AB - N1-(2-Alkoxyphenyl)piperazines additionally containing an N4-benzyl group bearing alcohol, amide, imide, or hydantoin functionalities were prepared and evaluated in the conditioned avoidance response (CAR) test predictive of clinical antipsychotic activity and in in vitro receptor-binding assays. Certain of the compounds display high affinity for the D2, 5-HT1A, and alpha 1-adrenergic receptors. Structures bearing acyclic amide, lactam, and imide functionalities display good biological activity, with a preference for the 1,3-disubstituted phenyl ring relative to the 1,4- and 1,2-congeners (7 vs 10 and 12). Every possible position of hydantoin attachment was investigated (e.g., substitution at N1, N3, and C5). The hydantoin involving attachment to N1 (24) was found to have good biological activity, whereas those hydantoins with attachment to N3 or C5 (22, 23, and 25) were inactive. Several of the smaller acetylated derivatives (30 and 33) have fair in vivo activity, which was lost in the case of the larger benzoyl analog 31. Uracil congener 34 had modest affinity for the D2 receptor (65 nM) as well as excellent in vivo activity. Benzylamino compounds display (viz. 27 and 35-38) moderate CAR activity but have surprising receptor affinity, often greater than those of comparable structures bearing a carbonyl (36 vs 7). Benzyl and benzhydryl alcohol compounds 40-48 are more active than amino structures 27 and 35-38 and also exhibit excellent in vivo activity in the CAR test with modest D2 and 5-HT1A receptor binding. PMID- 7473549 TI - Antifungal agents. 9. 3-Aryl-4-[alpha-(1H-imidazol-1-yl)arylmethyl]pyrroles: a new class of potent anti-Candida agents. AB - A new class of potent antifungal agents, namely, 3-aryl-4-[alpha-(1H-imidazol-1 yl)arylmethyl]-pyrroles, is described. These compounds are related to bifonazole and pyrrolnitrin, two compounds belonging to the class of antimycotic drugs. The synthesis of the title pyrroles has been performed starting from 1,3-diaryl-2 propen-1-ones, which were reacted with tosylmethyl isocyanide to give 3-aroyl-4 arylpyrroles. Reduction of the resulting compounds by lithium aluminum hydride furnished the related alcohols, which were treated with 1,1'-carbonyldimidazole to afford the required imidazole derivatives. Forty-four new pyrroles which incorporate an (arylmethyl)imidazole moiety in the 3-arylpyrrole structure were prepared by the above procedure and tested in vitro against Candida albicans and Candida spp. Among test compounds, 10 were found to be highly active against C. albicans. The most active derivative (27) was twice as potent (MIC90) as bifonazole, and its activity was 4 times greater than those of miconazole and ketoconazole. The other nine compounds showed antifungal activity of the same order of that of bifonazole and were ca. 2 times as active as miconazole and ketoconazole. Derivatives 21 and 27 tested in vivo against C. albicans A170 were shown to be highly effective in rabbit skin candidosis. Pharmacological studies on compounds 27 and other related pyrroles (19, 35, 36, 38, 39, and 49) are in progress to select one of them as a potential candidate for clinical experiments. PMID- 7473550 TI - Synthesis and antitumor activity of 4- and 5-substituted derivatives of isoquinoline-1-carboxaldehyde thiosemicarbazone. AB - Various substituted isoquinoline-1-carboxaldehyde thiosemicarbazones (12 compounds) have been synthesized and evaluated for antineoplastic activity in mice bearing the L1210 leukemia. Condensation of 4-bromo-1-methylisoquinoline (4) with ammonium hydroxide, methylamine, ethylamine, and N-acetylethylenediamine gave the corresponding 4-amino, 4-methylamino, 4-ethylamino, and 4-N (acetylethyl)amino derivatives, which were then converted to amides and subsequently oxidized to aldehydes followed by condensation with thiosemicarbazide to yield thiosemicarbazones 8a-c, 9a-c, and 16. Nitration of 4, followed by oxidation with selenium dioxide, produced aldehyde 18, which was then converted to the cyclic ethylene acetal 19. Condensation of 19 with morpholine followed by catalytic reduction of the nitro group and treatment with thiosemicarbazide afforded 5-amino-4-morpholinoisoquinoline-1-carboxaldehyde thiosemicarbazone (22). N-Oxidation of 1,5-dimethylisoquinoline, followed by rearrangement with acetic anhydride, gave, after acid hydrolysis, 1,5-dimethyl-4 hydroxyisoquinoline, which was converted to its acetate and then oxidized to yield 4-acetoxy-5-methylisoquinoline-1-carboxaldehyde (32). Sulfonation of 1,4 dimethylisoquinoline, followed by reaction with potassium hydroxide, acetylation, and oxidation, gave 5-acetoxy-4-methylisoquinoline-1-carboxaldehyde (40). Condensation of compounds 32 and 39 with thiosemicarbazide afforded the respective 4- and 5-acetoxy(5- and 4-methyl)thiosemicarbazones 33 and 40, which were then converted to their respective 4- and 5-hydroxy derivatives 34 and 41 by acid hydrolysis. The most active compounds synthesized were 4-aminoisoquinoline-1 carboxaldehyde thiosemicarbazone (9a) and 4-(methylamino)isoquinoline-1 carboxaldehyde thiosemicarbazone (9b), which both produced optimum % T/C values of 177 against the L1210 leukemia in mice when used at a daily dosage of 40 mg/kg for 6 consecutive days. Furthermore, when 9a was given twice daily at a dosage of 40 mg/kg for 6 consecutive days, a T/C value of 165 was obtained and 60% of the mice were 60-day long-term survivors. PMID- 7473552 TI - Microsphere formation in a series of derivatized alpha-amino acids: properties, molecular modeling, and oral delivery of salmon calcitonin. AB - A series of benzoylated and phenylsulfonylated amino acids are novel, low molecular weight, self-assembling molecules. At low pH, these compounds form microspheres that dissolve readily under neutral conditions. In a given synthetic series, those molecules with low aqueous solubility formed microspheres more readily than did the molecules possessing high water solubility, suggesting that the hydrophobicity of these compounds contributes to the ability to form microspheres. In addition, molecular modeling studies on selected compounds have shown that microsphere formation may depend also on various aromatic ring and dipole-dipole interactions, which could effect the extent and types of favorable stacking conformations between molecules. The microspheres prepared from these compounds have been used to effect the oral delivery of salmon calcitonin, a model protein drug, in both rodents and primates. PMID- 7473551 TI - 2-Substituted penems with amino acid-related side chains: synthesis and antibacterial activity of a new series of beta-lactam antibiotics. AB - A new series of 6-(hydroxyethyl)penems 2-substituted with amino acid-related side chains was synthesized. The nature of the amino acyl derivative proved to be crucial both from a synthetic point of view, as beta-lactam ring opening can compete with C-2 nucleophilic substitution, and for antibacterial activity. Primary amino acid amides emerged as the most suitable side chains for enhancing permeability through a Gram-negative outer membrane. In vitro activity of the new 2-[(aminoamido)methyl]penems 3a-u was influenced by the nature and position of the amide moiety, the ring size for cyclic amides, and the configuration of the amino acid. Compounds bearing amides derived from small N-methyl amino acids (such as 3a) or from cyclic amino acids (such as prolinamide 3p and 4 hydroxyprolinamide 3r) showed broad spectrum in vitro activity against both Gram positive and Gram-negative microorganisms. PMID- 7473553 TI - N-acylated alpha-amino acids as novel oral delivery agents for proteins. AB - A series of N-acylated alpha-amino acids were synthesized and shown to improve the oral delivery of two protein drugs, salmon calcitonin (sCT) and interferon alpha. Forty-five compounds in this series were tested in vivo in rats and primates. A significant positive correlation was found between the log P of the acylated amino acids and the decrease in serum calcium following oral dosage of sCT in rats. Such a correlation was not found for interferon-alpha. These derivatized amino acids only weakly inhibited the activity of trypsin or leucine aminopeptidase. Histological examinations of rat intestinal tissue after oral dosing of acylated amino acid/protein combinations revealed no detectable pathology. PMID- 7473554 TI - L-O-(2-malonyl)tyrosine: a new phosphotyrosyl mimetic for the preparation of Src homology 2 domain inhibitory peptides. AB - Inhibition of Src homology 2 (SH2) domain-binding interactions affords one potential means of modulating protein-tyrosine kinase-dependent signaling. Small phosphotyrosyl (pTyr)-containing peptides are able to bind to SH2 domains and compete with larger pTyr peptides or native pTyr-containing protein ligands. Such pTyr-containing peptides are limited in their utility as SH2 domain inhibitors in vivo due to their hydrolytic lability to protein-tyrosine phosphatases (PTPs) and the poor cellular penetration of the ionized phosphate moiety. An important aspect of SH2 domain inhibitor design is the creation of pTyr mimetics which are stable to PTPs and have reasonable bioavailability. To date, most PTP-resistant pTyr mimetics which bind to SH2 domains are phosphonates such as (phosphonomethyl)phenylalanine (Pmp, 2), [(monofluorophosphono)methyl]phenylalanine (FPmp, 3) or [(difluorophosphono)methyl]-phenylalanine (F2Pmp, 4). Herein we report the incorporation of a new non-phosphorus-containing pTyr mimetic, L-O-(2 malonyl)tyrosine (L-OMT, 5), into SH2 domain inhibitory peptides using the protected analogue L-N alpha-Fmoc-O'-(O",O"-di-tert-butyl-2-malonyl)tyrosine (6) and solid-phase peptide synthesis techniques. Five OMT-containing peptides were prepared against the following SH2 domains: the PI-3 kinase C-terminal p85 SH2 domain (Ac-D-(L-OMT)-V-P-M-L-amide, 10, IC50 = 14.2 microM), the Src SH2 domain (Ac-Q-(L-OMT)-E-E-I-P-amide, 11, IC50 = 25 microM, and Ac-Q-(L-OMT)-(L-OMT)-E-I-P amide, 14, IC50 = 23 microM), the Grb2 SH2 domain (Ac-N-(L-OMT)-V-N-I-E-amide, 12, IC50 = 120 microM), and the N-terminal SH-PTP2 SH2 domain (Ac-L-N-(L-OMT)-I-D L-D-L-V-amide, 13, IC50 = 22.0 microM). These results show that peptides 10, 11, 13, and 14 have reasonable affinity for their respective SH2 domains, with the IC50 value for the SH-PTP2 SH2 domain-directed peptide 13 being equivalent to that previously observed for the corresponding F2Pmp-containing peptide. OMT may afford a new structural starting point for the development of novel and useful SH2 domain inhibitors. PMID- 7473555 TI - Identification of efficient pentapeptide substrates for the tyrosine kinase pp60c src. AB - The development of inhibitors of protein tyrosine kinases (PTKs) is a promising approach to obtaining new therapeutic agents for a variety of diseases, particularly cancer. However, the discovery of peptide-based inhibitors has been hindered by the lack of small peptide substrate sequences (i.e. five residues or less) with which a variety of inhibitor designs could be readily evaluated by replacing the Tyr with natural and unnatural amino acids. These prototypical small peptide inhibitors could then form the basis for designing analogous conformationally constrained, peptide-mimetic or non-peptide inhibitors with improved therapeutic potential. In this study we have identified the best known small peptide substrate for the PTK pp60c-src, which is the parent of the src family of nonreceptor PTKs. This pentapeptide substrate, Ac-Ile-Tyr-Gly-Glu-Phe NH2, has a Km of 368 microM and Vmax of 1.02 mumol/min/mg when tested utilizing the assay methodology of Budde et al. (Anal. Biochem. 1992, 200, 347-351) after a series of modifications were made to more closely simulate the conditions inside a typical mammalian cell. This substrate was designed from information obtained by Songyang et al. (Nature 1995, 373, 536-539) with a 2.5 billion member combinatorial library of peptide substrates for pp60c-src. A second pentapeptide substrate, Ac-Glu-Asp-Ala-Ile-Tyr-NH2, with a weaker binding affinity (Km = 880 microM) but improved Vmax (1.86 mumol/min/mg), was also identified. This peptide was designed from the pp60c-src autophosphorylation sequence and information obtained by Songyang et al. (Ibid.) and Till et al. (J. Biol. Chem. 1994, 269, 7423-7428) with combinatorial libraries of peptide substrates. These new substrates provide sufficient binding affinities and rates of phosphorylation to be utilized for evaluating the relative effectiveness of various reversible and mechanism-based irreversible inhibitor designs for pp60c-src while appended to easily prepared small peptides. PMID- 7473556 TI - (+/-)-(N-alkylamino)benzazepine analogs: novel dopamine D1 receptor antagonists. AB - (+/-)-(N-Alkylamino)benzazepine analogs were prepared as novel dopamine D1 receptor antagonists to further elucidate the role of these receptor subtypes in the pharmacology and toxicology of cocaine. In the first series of compounds, (+/ )-7-chloro-8-hydroxy-3- [6-(N,N-dimethylamino)-hexyl]-1-phenyl-2,3,4,5-tetrahydro 1H-3-benzazepi ne (15) showed the highest affinity (Ki = 49.3 nM) and subtype selectivity for dopamine D1 over dopamine D2, 5-HT2a, and 5-HT2c receptors. Compounds 7a [(+/-)-7-Chloro-8-hydroxy-3-[4-(N,N-dimethylamino)butyl]-1-phenyl- 2,3,4,5-tetrahydro-1H-3-benzazepine], 11 [(+/-)-7-chloro-8-hydroxy-3-[6-[(N,N dimethylamino)hexyl]-1- phenyl-2,3,4,5-tetrahydro-1H-3-benzazepine-cyanoborane], and 15 were moderately potent dopamine D1 receptor antagonists as evidenced by their ability to block dopamine-stimulated adenylyl cyclase activity in rat caudate (predicted Ki values = 60, 34, and 21 nM, respectively). Compound 7a appears to be unique in that, despite its relatively potent inhibition of dopamine stimulated adenylyl cyclase, it demonstrated relatively weak binding affinity at the dopamine D1 receptors (Ki = 811 nM). Unlike previously reported N alkylbenzazepines, where a significant loss in dopamine D1 receptor binding affinity was observed when successive increases in the alkyl side chain size at the benzazepine nitrogen were made, several of these novel N-alkylamino analogs demonstrated high-affinity binding with an optimal chain length of six carbons. This initial series of compounds appears to be identifying another binding domain on the dopamine D1 receptor protein that has not previously been characterized and that accepts an amino function. Further, these compounds may serve as templates for the design of peripherally active dopamine D1 receptor antagonists. PMID- 7473557 TI - Improving the affinity and selectivity of a nonpeptide series of cholecystokinin B/gastrin receptor antagonists based on the dibenzobicyclo[2.2.2]octane skeleton. AB - We have recently described a novel series of nonpeptidic cholecystokinin-B (CCKB)/gastrin receptor antagonists based on a dibenzobicyclo[2.2.2]octane skeleton. We wish now to report on compounds arising out of our earlier work which have substantially greater affinity as antagonists for the CCKB/gastrin receptor system and which maintain, or improve on, the already high selectivity with respect to CCKA receptors. Thus, cis-7-[[[(1S)-[[3,5-dicarboxy phenyl)amino]carbonyl]-2- phenylethyl]amino]carbonyl]-8-[[(1 adamantylmethyl)amino]- carbonyl]-2,3:5,6-dibenzobicyclo[2.2.2]octane expressed a pKi of 8.80 in mouse cortical membranes at CCKB/gastrin receptors. The selectivity for these receptors over CCKA receptors was in the order of 1000 fold. PMID- 7473558 TI - A series of N4-imidoethyl derivatives of 1-(2,3-dihydro-1,4-benzodioxin-5 yl)piperazine as 5-HT1A receptor ligands: synthesis and structure-affinity relationships. AB - A series of unsubstituted and substituted succinimido, maleimido, and glutarimidoethyl derivatives of eltoprazine (3) was synthesized and tested for affinity for the 5-HT1A receptor in rat brain homogenates. The unsubstituted compounds have a moderate affinity for the receptor, while the affinity considerably increases by substitution at or enlargement of these cyclic ring systems. A good correlation was found between the inhibition constant Ki (expressed as pKi) and the lipophilicity (clogP). No correlation was observed between the pKi or pKi+ (local inhibition constant) and the basicity of the N4 nitrogen atom. PMID- 7473559 TI - Structure-activity relationships of cyclic pentapeptide endothelin A receptor antagonists. AB - Analogues of the natural product endothelin A (ETA) receptor antagonists cyclo(-D Trp1-D-Glu2-Ala3-D-Val4-Leu5-) (1) and cyclo(-D-Trp1-D-Glu2-Ala3-D-alloIle4-Leu5 ) (2) were prepared and tested for inhibitory activity against [125I]endothelin (ET-1) binding to protein ETA receptors. The DDLDL chirality sequence of the natural products appeared to be critical for inhibitory activity because conversion of either D-Trp or D-Glu (or both) in 1 to the corresponding L isomer(s) abolished this property. Systematic modifications at each position of the natural products clarified the structure-activity relationships and led to highly potent and selective ETA receptor antagonists. Most replacements of D-Trp1 and Leu5 with other amino acids caused a significant loss of inhibitory activity. In contrast, replacement of D-Glu2 with D-Asp2 enhanced the activity. With regard to the Ala3 position, all analogues with imino acids, independent of being cyclic or acyclic, showed higher affinities than did the amino acid analogues. In addition, most replacements with amino acids, which had various functional groups in their side chains, did not significantly modify ETA binding affinity. The D Val4/D-alloIle4 position was very important for inhibitory activity, and a beta position branched D-amino acid or a D-heteroarylglycine was preferable at this position. Among synthesized cyclic pentapeptides, compound 36 (BQ-518) was the most potent ETA receptor antagonist, with a pA2 of 8.1 against ET-1-induced vasoconstriction in isolated porcine coronary arteries. This compound also showed the greatest selectivity between ETA and ETB receptors (IC50 for human ETA = 1.2 nM, IC50 for human ETB = 55 microM). In contrast, compound 8 (BQ-123) is a highly soluble, potent, and selective ETA receptor antagonist (pA2 = 7.4, IC50 for human ETA = 8.3 nM, IC50 for human ETB = 61 microM). The sodium salt of 8 is practically freely soluble in saline. These compounds are useful tools for not only in vitro but also in vivo pharmacological studies. PMID- 7473561 TI - Mechanism of inhibition of poly(ADP-ribose) glycohydrolase by adenosine diphosphate (hydroxymethyl)pyrrolidinediol. AB - Adenosine diphosphate (hydroxymethyl)pyrrolidinediol (ADP-HPD), a nitrogen-in-the ring analog of ADP-ribose, was recently shown to be a potent and specific inhibitor of poly(ADP-ribose) glycohydrolase. Analysis of the inhibition kinetics of the hydrolase by ADP-HPD using the method of Lineweaver and Burk yields a noncompetitive double-reciprocal plot. Both the intercept (1/V) versus [inhibitor] replot and the slope (Km/V) versus [inhibitor] replot are hyperbolic, indicating partial noncompetitive inhibition. Inhibitor dissociation constants Kii = 52 nM and Kis = 80 nM were determined for ADP-HPD by analysis of the intercept versus [inhibitor] and slope versus [inhibitor] replots. These results show that although ADP-HPD is extremely potent in inhibiting poly(ADP-ribose) glycohydrolase, its effectiveness is limited by its partial inhibition. ADP-HPD was significantly less potent as an inhibitor of the NAD glycohydrolase from Bungarus fasciatus venom. Analysis of the inhibition kinetics using the Lineweaver and Burk method indicated that ADP-HPD was a linear-competitive inhibitor of the NAD glycohydrolase with a Ki of 94 microM. The results indicate that at low concentration ADP-HPD will be a selective inhibitor of poly(ADP ribose) glycohydrolase; however, complete inactivation of the activity will be difficult to obtain. PMID- 7473560 TI - Structure-taste correlations in sweet dihydrochalcone, sweet dihydroisocoumarin, and bitter flavone compounds. AB - The dihydrochalcone derivatives of the bitter flavonoids naringin and neohesperedin are intensely sweet. Phyllodulcin is as sweet as the dihydrochalcones with similar taste properties although its structure apparently resembles that of bitter flavanone or flavone. Multifaceted approaches, including X-ray crystal structure analysis, energy calculation, and structure comparison, have been employed to clarify the structure-taste correlations in these classes of compounds. In the crystal, naringin dihydrochalcone assumes a 'J'-shaped conformation with a fully-extended dihydrochalcone moiety while neohesperidin dihydrochalcone assumes the same overall conformation but with a partially extended moiety. A 2D conformational energy map of dihydrochalcone obtained using molecular mechanics revealed nine local minima. The pseudoequatorial and pseudoaxial forms of phyllodulcin have the same AM1 energies with a low energy barrier between them. The partially-extended form of dihydrochalcone and the pseudoequatorial form of phyllodulcin which are the maximally superposable conformers are proposed to be the active conformers. The major difference between the structures of flavone and phyllodulcin is not in the overall planarity but in the relative orientation of the pyrone and phenyl ring systems. PMID- 7473562 TI - Synthesis of isoornithines and methylputrescines. An evaluation of their inhibitory effects on ornithine decarboxylase. AB - 2-(Aminomethyl)-4-aminobutyric acid (isoornithine), 3-methylisoornithine, and 2,3 dimethylisoornithine were not decarboxylated by liver ornithine decarboxylase (ODC, EC 4.1.1.17) of thioacetamide-treated rats but were good competitive inhibitors of the enzyme (Ki ranged from 0.72 to 1.79 mM). When assayed in vivo in the treated rats, the above mentioned isoornithines were also found to inhibit liver ODC when administered 1 h before sacrifice. When the methylputrescines formally derived from the decarboxylation of several isoornithines were assayed on rat liver ODC, it was found that only 2,3-dimethylputrescine decreased the enzymatic activity. When assayed in vivo, it was found to decrease ODC activity by 60%, when the latter was measured 1 h after administration. The effect was reverted 4 h after administration of the drug. Isoornithines were not taken up by H-35 hepatoma cells; hence they did not affect their ODC activity. 2,3 Dimethylputrescine however, was transported into the cells and significantly decreased its ODC activity. PMID- 7473563 TI - Nitric oxide: a new paradigm for second messengers. PMID- 7473564 TI - Drug leads from combinatorial phosphodiester libraries. PMID- 7473565 TI - Synthesis and structure-activity relationships of substituted 1,4 dihydroquinoxaline-2,3-diones: antagonists of N-methyl-D-aspartate (NMDA) receptor glycine sites and non-NMDA glutamate receptors. AB - A series of mono-, di-, tri-, and tetrasubstituted 1,4-dihydroquinoxaline-2,3 diones (QXs) were synthesized and evaluated as antagonists at N-methyl-D aspartate (NMDA)/glycine sites and alpha-amino-3-hydroxy-5-methylisoxazole-4 propionic acid-preferring non-NMDA receptors. Antagonist potencies were measured by electrical assays in Xenopus oocytes expressing rat whole brain poly(A)+ RNA. Trisubstituted QXs 17a (ACEA 1021), 17b (ACEA 1031), 24a, and 27, containing a nitro group in the 5 position and halogen in the 6 and 7 positions, displayed high potency (Kb approximately 6-8 nM) at the glycine site, moderate potency at non-NMDA receptors (Kb = 0.9-1.5 microM), and the highest (120-250-fold) selectivity in favor of glycine site antagonism over non-NMDA receptors. Tetrasubstituted QXs 17d,e were more than 100-fold weaker glycine site antagonists than the corresponding trisubstituted QXs with F being better tolerated than Cl as a substituent at the 8 position. Di- and monosubstituted QXs showed progressively weaker antagonism compared to trisubstituted analogues. For example, removal of the 5-nitro group of 17a results in a approximately 100-fold decrease in potency (10a,b,z), while removal of both halogens from 17a results in a approximately 3000-fold decrease in potency (10v). In terms of steady-state inhibition, most QX substitution patterns favor antagonism at NMDA/glycine sites over antagonism at non-NMDA receptors. Among the QXs tested, only 17i was slightly selective for non-NMDA receptors. PMID- 7473566 TI - Enhanced D1 affinity in a series of piperazine ring substituted 1-piperazino-3 arylindans with potential atypical antipsychotic activity. AB - A study of the effect of aromatic substitution on D1 and D2 affinity in a series of previously reported trans-1-piperazino-3-phenylindans shows similar structure activity relationships for the two receptor sites. 6-Substituted derivatives have affinity for both receptors, and 6-chloro-or 6-fluoro-substituted derivatives show preference for D1 receptors. D1 affinity and selectivity are significantly increased in a series of new piperazine ring substituted derivatives. Potent D1 and D2 antagonism in vivo are confined to derivatives with relatively small substituents in the 2-position of the piperazine ring (e.g. 2-methyl,2,2 dimethyl, 2-spirocyclobutyl or 2-spirocyclopentyl). Consequently, the effect of aromatic substitution is examined in a series of 1-(2,2-dimethylpiperazino)-3 arylindans. All these compounds except the 4-, 5-, 7- and 4'-chloro-substituted derivatives have potent D1 affinity (IC50's below 10 nM) and the majority of the compounds antagonize SK&F 38393-induced circling in 6-OHDA-lesioned rats with ED50 values about 1 mumol/kg. In vitro all compounds show preference for D1 receptors, but in vivo they are equally effective as D1 and D2 antagonists. The compounds have high affinity for 5-HT2 receptors and selected compounds show high affinity for alpha 1 adrenoceptors. Furthermore, a subgroup consisting of (-)-38, (-)-39, (-)-41, and (-)-54 does not induce catalepsy in rats. These compounds have the potential of being "atypical" antipsychotics and have consequently been selected for further studies. The non-receptor-blocking enantiomers are shown to be inhibitors of DA and NE uptake in accordance with previous observations in compounds unsubstituted in the piperazine ring. Two compounds, (+)-38 and (+)-40, block DA uptake with IC50 values below 10 nM. Finally, the observed structure activity relationships are discussed in relation to previously published pharmacophore models for D2 and 5-HT2 receptors. It is concluded that the piperazine substituents might induce a different binding mode at the dopamine receptor sites, perhaps only at the D1 receptor site. PMID- 7473567 TI - Pyrrolo[2,1-c][1,4]benzothiazines: synthesis, structure-activity relationships, molecular modeling studies, and cardiovascular activity. AB - The synthesis and pharmacological evaluation of a series of pyrrolo[1,4]benzothiazine derivatives are described. These compounds, related to diltiazem, have been shown to be representative of a novel series of calcium channel antagonists. The IC50S for inhibition of [3H]nitrendipine binding calculated by radioreceptor assay on rat cortex and rat heart homogenates showed that some of the described compounds possess an affinity equal to or higher than those of the reference calcium antagonists verapamil and cis-(+)-diltiazem. Furthermore, the alteration of the benzothiazepinone system of diltiazem to the pyrrolo[1,4]benzothiazine system of the title compounds resulted in a clear-cut selectivity for cardiac over vascular tissue, as shown in functional studies. In fact comparison of calcium antagonist activity on guinea pig aorta strips with the negative inotropic activity, determined by using an isolated guinea pig left atrium, revealed that the compounds examined displayed higher selectivity than the reference standard, within a wide variation of data. A number of structure activity relationship trends have been identified, and possible explanation is advanced in order to account for the observed differences in selectivity. Prerequisite for in vitro calcium channel-blocking activity is the presence of two pharmacophores, namely, the substitution at C-4 and the substitution on the pyrrole ring. Two of the tested compounds, 8b and 28a, were identified as potent calcium antagonists selective for cardiac over vascular tissue. PMID- 7473568 TI - Synthetic and structure/activity studies on acid-substituted 2-arylphenols: discovery of 2-[2-propyl-3-[3-[2-ethyl-4-(4-fluorophenyl)-5- hydroxyphenoxy] propoxy]phenoxy]benzoic acid, a high-affinity leukotriene B4 receptor antagonist. AB - Structural derivatives of LY255283 have been studied as receptor antagonists of leukotriene B4. Substitution of the 2-hydroxyacetophenone subunit of 1 (LY255283) with a 2-arylphenol group provided entry into several new series that feature various mono- and diacidic core functionality. These new analogues, the subject of a broad structure-activity investigation, displayed significantly increased in vitro and in vivo activity as receptor antagonists of LTB4. A series of diaryl ether carboxylic acids demonstrated especially interesting activity and led to the discovery of compound 43b, 2-[2-propyl-3-[3-[2-ethyl-4-(4- fluorophenyl)-5 hydroxyphenoxy]-propoxy]phenoxy]benzoic acid (LY293111), a 2-arylphenol substituted diaryl ether carboxylic acid which displayed potent binding to human neutrophils (IC50 = 17 +/- 4.6 nM) and guinea pig lung membranes (IC50 = 6.6 +/- 0.71 nM), inhibition of LTB4-induced expression of the CD11b/CD18 receptor on human neutrophils (IC50 = 3.3 +/- 0.81 nM), and inhibition of LTB4-induced contraction of guinea pig lung parenchyma (pKB = 8.7 +/- 0.16). In vivo, 43b demonstrated potent activity in inhibiting LTB4-induced airway obstruction in the guinea pig when dosed by the oral (ED50 = 0.40 mg/kg) or intravenous (ED50 = 0.014 mg/kg) routes. A specific LTB4 receptor antagonist, 43b had little effect on inhibiting contractions of guinea pig lung parenchyma induced by leukotriene D4 (LTD4), histamine, carbachol, or U46619. Compound 43b has been chosen as a clinical candidate and is currently in phase I studies for a variety of inflammatory diseases. PMID- 7473569 TI - A 3-amino-4-hydroxy-3-cyclobutene-1,2-dione-containing glutamate analogue exhibiting high affinity to excitatory amino acid receptors. AB - The syntheses of several novel N-(hydroxydioxocyclobutenyl)-containing analogues of gamma-amino-butyric acid and L-glutamate were undertaken to test the hypothesis that derivatives of 3,4-dihydroxy-3-cyclobutene-1,2-dione (squaric acid), such as 3-amino-4-hydroxy-3-cyclobutene-1,2-dione, could serve as a replacement for the carboxylate moiety in neurochemically interesting molecules. The syntheses were successfully accomplished by preparation of a suitably protected diamine or diamino acid followed by reaction with diethyl squarate. Subsequent deprotection resulted in the isolation of the corresponding N (hydroxydioxocyclobutenyl)-containing analogues 13, 14, and 18. These analogues were screened as displacers in various neurochemical binding site assays. The L glutamate analogue 18, which showed high affinity as a displacer for kainate and AMPA binding, was also examined for agonist potency for CA1 pyramidal neurons of the rat hippocampal slice preparation. It rivaled AMPA as one of the most potent agonists for depolarizing pyramidal neurons in medium containing 2.4 mM Mg+2 ions in which kainate/AMPA receptors are active but NMDA receptors are inhibited (IC50 = 1.1 microM). It was 1 order of magnitude less potent for depolarizing pyramidal neurons under conditions in which kainate/AMPA receptors were inhibited by 10 microM CNQX but NMDA receptors were active in 0.1 mM Mg(+2)-containing medium (IC50 = 10 microM). Compound 18 did not induce sensitization of CA1 pyramidal cells to depolarization by phosphonate analogues of glutamate (the QUIS-effect). PMID- 7473570 TI - Kynurenic acid derivatives inhibit the binding of nerve growth factor (NGF) to the low-affinity p75 NGF receptor. AB - The ability of a series of substituted kynurenic acids, thienopyridinonecarboxylic acids, and related compounds to inhibit the binding of nerve growth factor (NGF) to the p75 NGF receptor (NGFR) was evaluated in a radioligand binding assay that utilized a biotinylated derivative of the extracellular domain of p75 NGFR (p75ext) fixed to streptavidin-coated plastic wells. Two compounds, 6-aminokynurenic acid (5h) and the 3-methyl ester of 4,7 dihydro-2-methyl-7-oxothieno[3,2-b]pyridine-3,5-dicarboxylic acid (16), were found to inhibit the binding of [125I]NGF to p75ext with IC50 values in the low micromolar range. Other amino-substituted kynurenic acids also possessed activity at slightly higher concentrations. Several structural features seem to be essential, including the carboxylic acid, a polar group on the benzene ring (or thiophene ring, in the case of analogues of 16), and the C-4 carbonyl group in the pyridinone ring. These compounds were also found to inhibit the binding of [125I]NGF to its receptors in membranes from PC12 cells (which express p75 as well as trka receptors for NGF) and DG44-CHO cells (transfected with full length p75 NGFR). The available data for 5h and 16 do not allow the determination of whether the effects of these compounds are mediated by their interaction with NGF or the NGF receptors. PMID- 7473571 TI - Structure-activity study of tripeptide thrombin inhibitors using alpha-alkyl amino acids and other conformationally constrained amino acid substitutions. AB - In our continuing effort to design novel thrombin inhibitors, a series of conformationally constrained amino acids (e.g. alpha-alkyl, N-alkyl cyclic, etc.) were utilized in a systematic structure-activity study of the P3, P2, and P1 positions of tripeptide arginal thrombin inhibitors. Early examples of this effort include: D-MePhe-Pro-Arg-H (15), Boc-D-Phg-Pro-Arg-H (18), D-1-Tiq-Pro-Arg H (23, D-1-Tiq = D-1,2,3,4-tetrahydroisoquinolin-1-ylcarbonyl), and Boc-D-Phe-Pro Arg-H (25).10a,20 The current work clarifies the contribution of each residue of the tripeptide arginals toward the potent and selective inhibition of thrombin relative to that of t-PA and plasmin. The alpha-methylarginal modification in the P1 residue resulted in analogs 30 (D-MePhe at P3) and 32 (D-1-Tiq at P3) which had lower potency toward thrombin while exhibiting improved selectivity. Analogs modified at the P2 site were found to be very sensitive to the conformational changes induced by variations in side chain ring size with the flexible pipecolinic acid 31 being 2 orders of magnitude less potent at thrombin inhibition than the conformationally constrained azetidine analog 20. Examination of the P3 binding region indicated that alpha-alkylphenylglycine residues resulted in a tendency to exhibit substantial improvements in selectivity over the nonalkylated residues. Combinations of optimal P3 and P2 changes led to compounds TFA-D-Phg(alpha Et)-Azt-Arg-H (16), TFA-D-Phg(alpha Me)-Azt-Arg-H (17), Ac-D-Phg(alpha Me)-Azt-Arg-H (21), TFA-D-Phg(alpha Me)-Pro-Arg-H (27), 30, and 32, which are clearly more selective for thrombin versus plasmin than the nonconformationally constrained compounds. PMID- 7473572 TI - Quantitative structure-activity relationships (QSARs) of N-terminus fragments of NK1 tachykinin antagonists: a comparison of classical QSARs and three-dimensional QSARs from similarity matrices. AB - The ability of three-dimensional quantitative structure-activity relationships (QSARs) derived from classical QSAR descriptors and similarity indices to rationalize the activity of 28 N-terminus fragments of tachykinin NK1 receptor antagonists was examined. Two different types of analyses, partial least squares and multiple regression, were performed in order to check the robustness of each derived model. The models derived using classical QSAR descriptors lacked accurate quantitative and predictive abilities to describe the nature of the receptor-inhibitor interaction. However models derived using 3D QSAR descriptors based on similarity indices were both robust and significantly predictive. The best model was obtained through the statistical analysis of molecular field similarity indices (n = 28, r2 = 0.846, r(cv)2 = 0.737, s = 0.987, PRESS = 7.102) suggesting that electronic and size-related properties are the most relevant in explaining the affinity data of the training set. The overall quality and predictive ability of the models applied to the test set appear to be very high, since the predicted affinities of three test compounds agree with the experimentally determined affinities obtained subsequently within the experimental error of the binding data. PMID- 7473574 TI - Synthesis of substituted 1-norbornylamines with antiviral activity. AB - The reaction of (+/-)-camphor (7) with triflic anhydride (Tf2O) yields the bridgehead triflate 8. The Nametkin rearrangement of 8 to 3 was realized by treatment with triflic acid (TfOH). The solvolysis of the bridgehead triflates 3 and 8 in acetonitrile affords the N-acetyl-1-norbornylamines 4 and 9. The Pd(0) catalyzed hydrogenation of 4 and 9 gives the amides 5 and 10. The corresponding 1 norbornylamines 2 and 13 and the N-ethyl derivatives 1, 6, 11, and 12 were obtained by basic hydrolysis or reduction with LiAlH4, respectively, of the amides 4, 5, 9, and 10. The antiviral activity of the hydrochlorides of some of the obtained 1-norbornylamines was evaluated against influenza A, herpes simplex 2, and African swine fever virus. Particularly noticeable is the activity of the hydrochlorides of 1 and 11 against influenza A virus (SI (selectivity index) = 1000). PMID- 7473573 TI - Structure-based design of nonpeptidic HIV protease inhibitors from a cyclooctylpyranone lead structure. AB - Recently, the novel cyclooctylpyranone HIV protease inhibitor 1 was identified in our labs, and an X-ray structure of this inhibitor complexed with HIV-2 protease was obtained. This crystal structure was used to develop two strategies for creating derivatives of 1 with enhanced enzyme inhibitory activity. The first strategy, substitution on the cyclooctyl ring, met with limited success, but provided some interesting information about the conformationally-flexible cycloocytyl ring on the inhibitors. The second strategy, substitution at the meta position of the aryl ring, was far more successful and generated compounds, such as the carboxamide derivatives 41 (Ki = 3.0 +/- 0.4 nM) and 36 (Ki = 4.0 +/- 0.8 nM), which were significantly more active than the corresponding unsubstituted cycloocytlpyranone 2 (Ki = 11.7 +/- 4.7 nM). An X-ray crystal structure of 36 complexed with HIV-1 protease indicated the increase in binding affinity is most likely due to the additional interactions between the amide substituent and the S3 region of the protease. PMID- 7473575 TI - The synthesis, structure-activity, and structure-side effect relationships of a series of 8-alkoxy- and 5-amino-8-alkoxyquinolone antibacterial agents. AB - A series of 1-cyclopropyl-6-fluoro-8-alkoxy (8-methyoxy and 8-ethoxy)-quionoline 3-carboxylic acids and 1-cyclopropyl-5-amino-6-fluoro-8-alkoxyquinoline-3 carboxylic acids has been prepared and evaluated for antibacterial activity. In addition, they were also compared to quinolones with classic substitution at C8 (H, F, Cl) and the naphthyridine nucleus in a phototoxicity and mammalian cell cytotoxicity assay. The series of 8-methoxyquinolones had antibacterial activity against Gram-positive, Gram-negative, and anaerobic bacteria equivalent to the most active 8-substituted compounds (8-F and 8-Cl). There was also a concomitant reduction in several of the potential side effects (i.e., phototoxicity and clonogenicity) compared to the most active quinolones with classic substitution at C-8. The 8-ethoxy derivatives had an even better safety profile but were significantly less active (2-3 dilutions) in the antibacterial assay. PMID- 7473576 TI - Hypoxia-selective agents derived from 2-quinoxalinecarbonitrile 1,4-di-N-oxides. 2. AB - Hypoxic cells are an important target for antitumor therapy because tumors are typically characterized by such cells. Virtually all tumors which are present as solid masses contain hypoxic cells, while normal cells generally have an adequate supply of oxygen. Accordingly, antitumor agents can be made selective for tumors by virtue of high activity under hypoxic conditions. The initial purpose of this work was to determine the influence of different groups in position 3. Thus, the synthesis of some 3-NH-substituted derivatives (2a, 3a, 4a) starting from 3-amino 2-quinoxalinecarbonitrile 1,4-di-N-oxide (1a) is described. Reductive deamination of compounds 1a-k provides the 2-quinoxalinecarbonitriles 5a-k, which are more potent, while selectivity is maintained or increased in some derivatives. The compound 7-(4-nitrophenyl)-2-quinoxalinecarbonitrile 1,4-di-N-oxide (5k) is 150 fold more potent than tirapazamine (3-amino-1,2,4- benzotriazine 1,4-di-N-oxide), which has been used as a standard. Three derivatives (5g,i,k) show a hypoxic cytotoxicity ratio (HCR) > or = 200, better than that of tirapazamine (HCR = 75) in V79 cells. Replacement of the 3-amino group by chlorine affords the potent but nonselective 3-chloro derivatives 6a-k showing similar toxicities under both aerobic and hypoxic conditions. These compounds were used as intermediates for the synthesis of a new series of water-soluble compounds derived from 3-[[(N,N- dialkylamino)alkyl[amino]-2-quinoxalinecarbonitrile 1,4-di-N-oxides 10a-i and 11a i. The 7-chloro and the 7-trifluoromethyl derivatives 10b,f have demonstrated high potency (0.4 and 0.3 microM) and excellent selectivity (HCR = 250 and 340). Several 7-chloro analogues, 12b, 13b.1,b.2, and 14b, and the dimer 16b have been prepared and evaluated in order to determine the optimum lateral chain in position 3, which appears to be the [(N,N-dimethylamino)propyl]amino moiety. PMID- 7473577 TI - 5-Arylthio-substituted 2-amino-4-oxo-6-methylpyrrolo[2,3-d]pyrimidine antifolates as thymidylate synthase inhibitors and antitumor agents. AB - Classical antifolate inhibitors of thymidylate synthase (TS) often require the reduced folate uptake system in order to exert their antitumor effects. In addition, these analogues are polyglutamylated via the enzyme folylpoly-gamma glutamate synthetase (FPGS), which prevents analogue efflux from the cell and usually increases their inhibitory potency against TS. Impaired function of the reduced folate uptake system and that of FPGS are potential sources of resistance to such antifolates. We designed and synthesized a classical 6-5 ring-fused analogue N-[4-[(2-amino-6-methyl-3,4-dihydro-4-oxo-7H-pyrrolo[2,3- d]pyrimidin-5 yl)thio]-benzoyl]-L-glutamic acid (5) and a nonclassical 6-5 ring-fused analogue 2-amino-6-methyl-5-(pyridin-4-ylthio)-3,4-dihydro-4-oxo-7H-pyrrolo [2,3- d]pyrimidine (6) as TS inhibitors and antitumor agents. The syntheses of analogues 5 and 6 were achieved via the oxidative addition of the sodium salt of ethyl 4-mercaptobenzoate or 4-mercaptopyridine to 2-(pivaloylamino)-6-methyl-3,4 dihydro-4-oxo-7H-pyrrolo[2,3-d]pyri midine (17) in the presence of iodine. For the synthesis of 5 the ester obtained from the reaction was deprotected and coupled with diethyl L-glutamate followed by saponification. Compound 5 was a potent inhibitor of human and bacterial TS with IC50 values of 42 and 21 nM, respectively. Compound 6 was 10-fold less potent than 5 against human TS but more than 4700-fold less potent than 5 against Lactobacillus casei TS. The classical analogue 5 was neither a substrate nor an inhibitor of human FPGS derived from CCRF-CEM cells. Compound 5 was cytotoxic to CCRF-CEM and FaDu tumor cell lines as well as to an FPGS-deficient subline of CCRF-CEM. Thymidine protection studies established that TS was the primary target of 5. PMID- 7473578 TI - Topoisomerase II-mediated DNA cleavage by adocia- and xestoquinones from the Philippine sponge Xestospongia sp. AB - Investigation of an orange Xestospongia sp. sponge collected at Cape Bolinao in northern Luzon, Philippines, yielded the known compounds adociaquinones A and B (1, 2) and six new metabolites, secoadociaquinones A and B (3, 4), 14 methoxyxestoquinone (5), 15-methoxyxestoquinone (6), 15-chloro-14 hydroxyxestoquinone (7), and 14-chloro-15-hydroxyxestoquinone (8). All compounds showed inhibition of topoisomerase II in catalytic DNA unwinding and/or decatenation assays. Furthermore, adociaquinone B showed activity in a KSDS assay, suggesting it inhibits the enzyme by freezing the enzyme-DNA cleavable complex. Interestingly, adociaquinone B did not displace ethidium bromide from DNA or unwind supercoiled DNA, implying it does not intercalate DNA. PMID- 7473579 TI - New orally active serine protease inhibitors: structural requirements for their good oral activity. AB - Synthesis and structural requirements for good oral activity of a series of para substituted benzoyl esters of 4-hydroxybenzamidine serine protease inhibitors are described. The structure required for good oral activity was found to be general formula II whose corresponding ester has to be hydrolyzed in the intestine before absorption through the mucous membranes or in plasma after absorption. Biological evaluation of oral absorption using plasma anti-trypsin activity was useful for rapid evaluation. By measuring their actual plasma concentrations after oral administration, compounds 14 and 16b were confirmed to show good area under the plasma concentration-time curves (AUC). Their plasma concentrations corresponded to their plasma anti-trypsin activity. Structure-oral activity relationships are discussed. PMID- 7473580 TI - Steroidal spiro-gamma-lactones that inhibit 17 beta-hydroxysteroid dehydrogenase activity in human placental microsomes. AB - The important enzyme 17 beta-hydroxysteroid dehydrogenase (17 beta-HSD) is known to regulate intracellular levels of biologically active steroids, namely, androgens and estrogens. In an effort to develop potent inhibitors of 17 beta-HSD for reducing the levels of active steroids, we found that steroidal spiro-gamma lactones inhibit 17 beta-HSD activity. In this report, we describe the synthesis of 11 spiro-gamma-lactone analogs containing a steroidal C-18 or C-19 nucleus and compare their relative inhibitory effects on 17 beta-HSD activity in the human placenta microsomes that catalyze the interconversion of androgens and estrogens. To void the interaction of the cytosolic 17 beta-HSD activity that is specific for the interconversion of estrone and estradiol, we used 4-androstenedione as substrate. Analysis of the inhibitory effect exerted by these analogs on microsomal 17 beta-HSD activity indicates that spiro-gamma-lactones containing the C-18 nucleus are more potent inhibitors than C-19 nucleus analogs. The best inhibition was obtained with the phenolic spiro-gamma-lactone 7 (3-hydroxy-19-nor 17 alpha-pregna-1,3,5(10)-triene 21,17-carbolactone), which has an IC50 value of 0.27 microM, and was much lower than the competitive effect of the unlabeled substrate 4-androstenedione, which has an IC50 value of 1.40 microM. Preincubation with lactone 7 did not inactivate 17 beta-HSD activity. The results thus suggest that lactone 7 is a reversible in inhibitor. Lactone 7 is selective for microsomal 17 beta-HSD activity, as no inhibition was observed for cytosolic 17 beta-HSD activity. PMID- 7473581 TI - 1 alpha,25-dihydroxyvitamin D3 analogs featuring aromatic and heteroaromatic rings: design, synthesis, and preliminary biological testing. AB - Aromatic compounds 2a-c, analogs of 1 alpha, 25-dihydroxyvitamin (calcitriol, 1), and heteroaromatic compounds 4a-c and 5a-c, analogs of 19-nor-1 alpha, 25 dihydroxyvitamin D3 (3), were designed to simulate the topology of their biologically potent parent compounds while avoiding previtamin D equilibrium. Convergent and facile total syntheses of the analogs (+)-2b, (+)-2c, (-)-4b, and (-)-5b were achieved via carbonyl addition of regiospecifically formed organolithium nucleophiles to the enantiomerically pure C,D-ring ketone (+)-17, characteristic of natural calcitriol (1). Likewise, hybrid analogs 20a-c were prepared to determine whether incorporation of a known potentiating side chain would lead to increased biological activity. Preliminary in vitro biological testing showed that aromatic analogs (+)-2b, (+)-2c, and 20a-c as well as heteroaromatic analogs (-)-4b and (-)-5b have very low affinities for the calf thymus vitamin D receptor but considerable antiproliferative activities in murine keratinocytes at micromolar concentration. No biological advantage was observed in this keratinocyte assay for the doubly modified hybrid analogs 20a-c over the singly modified parent (+)-2b. Analog (+)-2b, but surprisingly not the corresponding analog 20b differing from (+)-2b only in the side chain, showed considerable activity in nongenomic opening of calcium channels in rat osteosarcoma cells. PMID- 7473582 TI - Thiopyranol[2,3,4-c,d]indoles as inhibitors of 5-lipoxygenase, 5-lipoxygenase activating protein, and leukotriene C4 synthase. AB - The attachment of an arylacetic or benzoic acid moiety to the thiopyrano[2,3,4 c,d]indole nucleus results in compounds which are highly potent and selective 5 lipoxygenase (5-LO) inhibitors. These compounds are structurally simpler than previous compounds of similar potency in that they contain a single chiral center. From the data presented, 2-[[1-(3-chlorobenzyl)-4-methyl-6-[(5 phenylpyridin-2-yl)methoxy]- 4, 5-dihydro-1H-thiopyrano[2,3,4-c,d]indol-2 yl]methoxy]-phenylacetic acid, 14b, was shown to inhibit 5 hydroperoxyeicosatetraenoic acid (5-HPETE) production by human 5-LO (IC50 of 18 nM). The acid 14b is highly selective as an inhibitor of 5-LO activity when compared to the inhibition of ram seminal vesicle cyclooxygenase (IC50 > 5 microM) or human leukocyte leukotriene A4 (LTA4) hydrolase (IC50 > 20 microM). In addition, 14b was inactive in a 5-lipoxygenase-activating protein (FLAP) binding assay at 10 microM. In vivo studies showed that 14b is bioavailable in rat and functionally active in the hyperreactive rat model of antigen-induced dyspnea (74% inhibition at 0.5 mk/kg po; 2 h pretreatment). In the conscious squirrel monkey model of asthma, 14b showed excellent functional activity at 0.1 mg/kg against antigen-induced bronchoconstriction (94% inhibition of the increase in RL and 100% inhibition in the decrease in Cdyn; n = 4). Resolution of this compound gave (-)-14b, the most potent enantiomer (IC50 = 10 nM in the human 5-LO assay), which was shown to possess the S configuration at the chiral center by X-ray crystallographic analysis of an intermediate. Subsequent studies on the aryl thiopyrano[2,3,4-c,d]indole series of inhibitors led to the discovery of potent dual inhibitors of both FLAP and 5-LO, the most potent of which is 2-[[1-(4 chlorobenzyl)-4-methyl-6-(quinolin-2-ylmethoxy)-4, 5-dihydro-1H-thiopyrano[2,3,4 c,d]indol-2-yl]methoxy]phenylacetic acid, 19. Acid 19 has an IC50 of 100 nM for the inhibition of 5-HPETE production by human 5-LO and is active in a FLAP binding assay with an IC50 of 32 nM. Furthermore, thiopyrano[2,3,4-c,d]indoles such as 1 and 14b are capable of inhibiting the LTC4 synthase reaction in a dose dependent manner (IC50s of 11 and 16 microM, respectively, compared to that of LTC2 at 1.2 microM) in contrast to other, structurally distinct 5-LO inhibitors. It has also been observed that the thiopyrano[2,3,4-c,d]indole class of compounds strongly promotes the translocation of 5-LO from the cytosol to a membrane fraction in the presence or absence of the ionophore A23187.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 7473583 TI - Neurosteroid analogues. 3. The synthesis and electrophysiological evaluation of benz[e]indene congeners of neuroactive steroids having the 5 beta-configuration. AB - A series of 7-(2-hydroxyethyl)benz[e]indene analogues of 3 alpha-hydroxy-5 beta pregnan-20-one (7), a neuroactive steroid known to be a positive allosteric modulator of GABAA receptor function, was prepared. Electrophysiological measurements carried out on cultured rat hippocampal neurons were used to evaluate the modulatory effects of the analogues on GABAA receptor function. Analogues were tested for their ability to potentiate 1 microM GABA-mediated chloride currents and for their ability to directly gate chloride currents at this ligand-gated ion channel. Active analogues typically enhanced GABA-mediated currents at concentrations below those required to directly gate chloride currents. The dose-response relationships for potentiation of 1 microM GABA mediated chloride currents were studied for [3S-(3 alpha, 3a alpha, 5a beta, 7 beta, 9a alpha, 9b beta)]-1- [dodecahydro-7-(2-hydroxyethyl)-3a-methyl-1H benz[e]inden-3- yl]ethanone (3), steroid 7, 3 alpha-hydroxy-5 alpha-pregnan-20 one (5), and the analogous 7 alpha-(2-hydroxyethyl)benz[e]indene analogue of steroid 5 (compound 1). Compound 3 was the most active potentiator (EC50 = 0.017 microM) of GABA-mediated current. The direct gating actions of compound 3 were not observed at a concentration of 1 microM, but were observed at a concentration of 10 microM. PMID- 7473584 TI - alpha-Hydroxy phosphinyl-based inhibitors of human renin. AB - The design and application of alpha-hydroxy phosphonates, a new class of transition state analogs, toward the discovery of novel and potent inhibitors of the aspartyl protease renin is described. Tripeptidic alpha-hydroxy diethyl phosphonate 3, the first example in this series, was found to be a good inhibitor of human renin (IC50 = 29 nM), and preliminary studies led to the choice of alpha hydroxy dimethyl phosphonate 15 (IC50 = 16 nM) as a base-line compound for further structure-activity relationship study. Corresponding phosphinate (28-30) and phosphine oxide (23 and 24) analogs of 15 were prepared to assess the steric and electronic requirements around the phosphorus center. Evaluation of these analogs suggested that the presence of at least one alkoxy group on phosphorus was a critical requirement for good activity. Inhibitors with leucine at P2 possessed better in vitro activity than the corresponding P2 histidine analogs (15, IC50 = 16 nM vs 37, IC50 = 220 nM; 33, IC50 = 8.5 nM vs 40, IC50 = 41 nM). Compound 34 (IC50 = 31 nM), the P3 aminocaproic analog of 15, showed complete and long-lasting inhibition of plasma renin activity while eliciting a 10-15 mmHg drop in mean arterial pressure when administered intravenously at 1 mumol/kg in conscious, sodium-depleted, cynomolgus monkeys. In summary, the alpha-hydroxy phosphonates represent a promising and structurally novel class of transition state analog inhibitors of human renin. PMID- 7473585 TI - 1,2-Diarylcyclopentenes as selective cyclooxygenase-2 inhibitors and orally active anti-inflammatory agents. AB - A series of 1,2-diarylcyclopentene methyl sulfones and sulfonamides have been shown to be remarkably potent and selective cyclooxygenase-2 (COX-2) inhibitors. The methyl sulfone analogs 7 showed excellent COX-2 activity, with IC50s ranging from 0.003 (7f,n) to 0.87 (7o) microM. In addition, most analogs of 7 showed no activity (IC50 > 100 microM) against the COX-1 enzyme. Replacement of the methyl sulfone moiety with a sulfonamide group gave a slightly more potent (typically 2 5-fold) but less selective COX-2 inhibitor, mainly due to an increase (20- > 100 fold) in COX-1 activity. However, in vitro COX-1/COX-2 selectivity for the sulfonamides 8 could be increased in many cases by simply incorporating a substituent at the 3-position of the phenyl group. Furthermore, in vitro selectivity increased with the size and number of substituents, as demonstrated in the selectivity trend of 8k (8000) > 8j (1900) > 8i (500) > 8h (100). More importantly, the sulfonamide COX-2 inhibitors showed greatly enhanced oral activity in the rat model of established adjuvant-induced arthritis, with inhibition values of 79.0% (8a), 81.5% (8c), and 83.0% (8g) at 1 mg/kg. On the basis of its overall biological profile, sulfonamide 8c was evaluated as a potential clinical candidate, displaying an ED50 of 22 mpk in the rat carrageenan induced paw edema model and an ED50 of 0.16 mpk in the rat established adjuvant induced arthritis model with no indication of gastrointestinal toxicity in rats and mice at 200 mpk. In addition, a preparative-scale synthetic route to sulfonamide 8c has been developed. PMID- 7473586 TI - Y1 and Y2 receptor selective neuropeptide Y analogues: evidence for a Y1 receptor subclass. AB - Neuropeptide Y (NPY), a 36-residue polypeptide produced abundantly in both nervous and peripheral tissues, appears to play a significant role in the regulation of diverse biological processes, including feeding behavior and cardiovascular and psychotropic functions. The actions of NPY are mediated through effective binding to specific receptors of which two, designated Y1 and Y2, have been well characterized. A shortened cyclic analogue of NPY, des-AA10-17 cyclo-7/21[Cys7,21]NPY, was shown to retain high affinity for both human neuroblastoma SK-N-MC and SK-N-BE2 cell types (expressing Y1 and Y2 receptors, respectively). Increasing the size of the ring (des-AA10-17-cyclo 2/27[Cys2,27]NPY) in the present study produced a high-affinity analogue (Ki = 3.0 vs 0.3 nM for NPY) that bound exclusively to Y2 receptors. Using the feedback from structure-activity relationships, we also describe the optimization of specific substitutions and bridging arrangements leading to the production of other truncated, high-affinity Y1 selective analogues which bind, as does NPY itself, in the low-nanomolar range. Of greatest significance, des-AA10-17-cyclo 7/21[Cys7,21,Pro34]NPY (11) was found to possess agonistic properties with an affinity comparable to that of the native NPY molecule when tested for its ability to inhibit norepinephrine-stimulated cAMP release in SK-N-MC human neuroblastoma cells. Compound 11 also caused an increase in blood pressure in anesthetized rats. However, in two central nervous system models of Y1 receptor function, stimulation of feeding and anxiolytic activity, this analogue was inactive, which suggests the presence of a new subclass of receptors. In summary, the present results demonstrate that residues 10-17 of NPY are not directly involved in either Y1 or Y2 receptor recognition or activation. This suggests that the selectivity of NPY receptors is highly dependent on subtle conformational changes such as the substitution of residue 34 to a proline or the introduction of intramolecular constraints. Additionally, we have produced an analogue of NPY that selectively activates peripheral NPY Y1 receptors. PMID- 7473587 TI - Structure-activity relationships of cytotoxic cholesterol-modified DNA duplexes. AB - Short DNA duplexes with cholesterol linked at the 3'-terminus of each strand have unique, selective cytotoxic properties. The structural requirements for biological activity were explored through chemical synthesis of analogs and testing in cultured hepatoma cells. Effects of modifications to the sequence, backbone, 3'-sterol, 3'-linker, and 5'-terminus were evaluated. Self complementary 3'-modified oligodeoxynucleotide (ODN) 10-mers were prepared from solid supports bearing the modification and linker of interest. Any changes to the normal phosphodiester backbone were poorly tolerated. The presence of cholesterol or a closely related sterol was an absolute requirement for activity. The length and position of attachment of the linker to cholesterol was important, with longer linkers showing reduced activity. Large, lipophilic groups at the 5' terminus gave reduced cytotoxicity and poor solubility properties. The short length and unique structure of these ODNs allowed efficient automated synthesis on a 400 mumol scale and simplified purification. PMID- 7473589 TI - Prospects for improved antidepressants. PMID- 7473588 TI - Inhibition of E-selectin-, ICAM-1-, and VCAM-1-mediated cell adhesion by benzo[b]thiophene-, benzofuran-, indole-, and naphthalene-2-carboxamides: identification of PD 144795 as an antiinflammatory agent. AB - It was previously reported that 3-alkoxybenzo[b]thiophene-2-carboxamides exemplified by 1, 5-methoxy-3-(1-methylethoxy)benzo[b]thiophene-2-carboxamide, decreased the adherence of neutrophils to activated endothelial cells by inhibiting the upregulation of the adhesion molecules E-selectin and ICAM-1 on the surface of the endothelium. This finding is extended here to a series of 3 thiobenzo[b]thiophene-2-carboxamides and also heterocyclic analogs of 1, including benzofurans, indoles, and napthalenes. The compounds that inhibited the expression of E-selectin and ICAM-1 had the same effect on the expression of VCAM 1. PD 144795, 5-methoxy-3-(1-methylethoxy)benzo[b]thiophene-2-carboxamide 1-oxide (44), the sulfoxide analog of 1, was orally active in several models of inflammation. The in vitro and in vivo activity of PD 144795 resided predominately in the S-enantiomer. PMID- 7473590 TI - 1-(1-[4-[(N-acetyl-4-piperidinyl)oxy]-2-methoxybenzoyl]piperidin-4- yl)-4H-3,1 benzoxazin-2(1H)-one (L-371,257): a new, orally bioavailable, non-peptide oxytocin antagonist. PMID- 7473591 TI - A novel strategy for improving ligand selectivity in receptor-based drug design. AB - A major desirable characteristic of many drugs is their ability to interact specifically with only one variety of the target receptor among many others. It is remarkable that, even when accurate three dimensional structures for the target biomolecules are available, there is no well-established methodology to describe their differences and use them for the design of selectively-interacting compounds. This work presents a novel method that uses multivariate GRID descriptors and principal component analysis (PCA) with the aim of revealing the most relevant structural and physicochemical differences between biomacromolecules related to receptor selectivity. The methodology is described through an example involving the study of bacterial (Escherichia coli) and recombinant human varieties of the dihydrofolate reductase (EC 1.5.1.3, DHFR) enzyme. This analysis easily unveils the most important regions on these biomolecules which should be taken into consideration for the design of selectively interacting compounds. PMID- 7473592 TI - Design, synthesis, and structure-activity relationship of novel dinucleotide analogs as agents against herpes and human immunodeficiency viruses. AB - A new acyclic nucleoside phosphonate (13) containing an adenine moiety was synthesized, which acted as an excellent inhibitor of calf mucosal adenosine deaminase. This inhibitory property allows it to exert great synergistic effect on certain antiviral agents (e.g., ara-A, 37). Phosphonate 13 was not phosphorylated by the bovine brain guanylate kinase nor by 5-phosphoribosyl 1 pyrophosphate synthetase. Syntheses of biologically active nucleotide phosphonate 40 and its phosphonoamidate derivative 42 were accomplished, which showed remarkable activity against herpes viruses and exhibited low host cell toxicity. 3'-Azido-nucleoside phosphonate 20 and 3'-fluoronucleoside phosphonate 32, as well as the corresponding dinucleotide analogs 47 and 48, and their respective phosphonoamidates 53-56 were also synthesized as new compounds, among which phosphonoamidates 53-56 showed potent activity against human immunodeficiency virus. Phosphonoamidates 55 and 56 bearing a methyl D-alaninate moiety exhibited less cellular toxicity than 53 and 54 bearing a methyl L-alaninate moiety. Nucleotide phosphonate 40 as well as dinucleotide phosphonates 47 and 48 were found susceptible to degradation by phosphodiesterases. Their respective phosphonoamidates 42 and 53-56, however, were completely resistant to snake venom and spleen enzymes. PMID- 7473593 TI - Differences in backbone structure between angiotensin II agonists and type I antagonists. AB - Type I angiotensin II antagonists with O-methyl-L-homoserine [HSer(gamma-OMe)] and delta-methoxy-L-norvaline [Nva(delta-OMe)] at position 8 have been prepared by the solid-phase method, purified by reverse-phase HPLC, and bioassayed in the rat uterus, and their backbone conformational properties were investigated by nuclear Overhauser effect (NOE) spectroscopy. [Sar1,HSer-(gamma-OMe)8]ANGII, [HSer(gamma-OMe)8]ANGII, [Des1,HSer(gamma-OMe)8]ANGII, [Sar1,Nva(delta-OMe)8] ANGII, and [Des1,Nva(delta-OMe)8]ANGII had, respectively, the following antagonist activities, pA2: 7.6, 7.5, < 6.0, 7.1, and 6.9. Analogs of [Sar1]ANGII with delta-hydroxy-L-norvaline [Nva(delta-OH)], delta-methoxy-L-norvaline [Nva(delta-OMe)], 4'-carboxyphenylalanine [Phe(4'-COOH)], and 4' (trifluoromethyl)phenylalanine [Phe(4'-CF3)] at position 4 were also prepared by solid phase and bioassayed in the rat uterus. [Sar1,Nva(delta-OH)4]ANGII, [Aib1,Nva(delta-OMe)4]ANGII, [Sar1,DL-Phe(4'-COOH)4]ANGII, and [Sar1,DL-Phe(4' CF3)4]ANGII had, respectively, agonist activities as follows: 4%, 1.5%, 3%, < 0.1%, and < 0.1%. These data emphasize that replacement of Ile8 in Sarilesin with the higher homologs HSer(gamma-OMe) and Nva(delta-OMe) does not greatly alter the structural requirements necessary for expression of type I antagonist activity, while replacement of the tyrosine hydroxyl in [Sar1]ANGII by the carboxylate or the trifluoromethyl group abolishes activity, suggesting that the tyrosinate pharmacophore cannot be replaced by any negatively charged or electronegative group. Conformational investigation of the ANGII type I antagonists [HSer(gamma OMe)8]ANGII and [Sar1Nva(delta-OMe)8]ANGII in DMSO by 1D-NOE spectroscopy revealed that the Tyr-Ile-His bend, a conformational property found in ANGII and [Sar1]ANGII (J. Biol. Chem. 1994, 269, 5303) is not present in type I antagonists, providing for the first time an important conformational difference between angiotensin II agonists and type I antagonists. PMID- 7473594 TI - 4-(Heteroarylthio)-2-biphenylyltetrazoles as nonpeptide angiotensin II antagonists. AB - A series of 4-(heteroarylthio)-2-biphenylyltetrazoles was prepared, and the compounds were examined for their ability to displace [3H]AII from angiotensin II receptors. Analogues that exhibited significant receptor binding affinities at less than 10 microM were investigated further for potential antagonism of angiotensin II-mediated contraction of rabbit isolated aortic rings. Three 4 (heteroarylthio)-2-biphenylyltetrazoles were identified that exhibited sub micromolar angiotensin II receptor binding affinities. These compounds and two reference agents, saralasin and losartan (DUP-753), exhibited concentration dependent reversal of angiotensin II contraction in isolated aortic rings parallel to their receptor binding affinities. Molecular modeling studies were conducted to examine the conformational effects of the novel sulfide bridging unit contained in these 4-(heteroarylthio)-2-biphenylyltetrazoles. The biological effects of the sulfide bridge as well as alterations in the heteroaromatic moiety were investigated, and the resulting structure--activity relationships are discussed. PMID- 7473595 TI - A new series of pyridinone derivatives as potent non-nucleoside human immunodeficiency virus type 1 specific reverse transcriptase inhibitors. AB - 4-(Arylthio)-pyridin-2(1H)-ones variously substituted in their 3-, 5-, and 6 positions have been synthesized as a new series of 1-[(2-hydroxyethoxy)methyl]-6 (phenylthio)thymine (HEPT)-pyridinone hybrid molecules. Biological studies revealed that some of them show potent HIV-1 specific reverse transcriptase inhibitory properties. Compounds 16 and 7c, the most active ones, inhibit the replication of HIV-1 at 3 and 6 nM, respectively. PMID- 7473596 TI - A novel class of cyclic beta-dicarbonyl leaving groups and their use in the design of benzisothiazolone human leukocyte elastase inhibitors. AB - Human leukocyte elastase (HLE) has been proposed to be a primary mediator of pulmonary emphysema, and inhibitors of this enzyme should be effective in the treatment of emphysema and other pulmonary diseases. We have discovered a novel class of alicyclic and heterocyclic leaving groups which share one common structural feature, a cyclic beta-dicarbonyl. This design concept for leaving groups has not been previously reported. A structure-activity relationship has been developed and the concept extended to several types of alicyclic and heterocyclic beta-dicarbonyl systems. This work led to the identification of a potent (K*i of 0.066 nM) and tissue stable (in vitro: blood t1/2 = 160 min, liver t1/2 > 240 min) benzisothiazolone HLE inhibitor, WIN 65936 (13b). PMID- 7473597 TI - Inhibition of the EGF-stimulated cellular proliferation of ER 22 cells by hydroxybiphenyl derivatives. AB - Several series of hydroxybiphenyl compounds substituted by a hydrophobic group (tert-butyl or phenyl) and bearing a free or protected carboxylic moiety were synthesized. The compounds were tested for their ability to inhibit the intrinsic tyrosine protein kinase activity of the EGF-receptor in vitro and the EGF stimulated DNA-synthesis by ER 22 cells. Although the compounds of each series had poor in vitro inhibitory potencies (IC50 >> 100 microM), most of them inhibited the EGF-dependent cellular proliferation of ER 22 cells at relatively low doses (IC50 = 1.1 microM for compound 14). Structure-activity studies based on the cellular results showed that the most interesting series was the linear terphenyl series B of 2'-hydroxy-1,1':4',1"-terphenyl-4-carboxylates. The availability of the hydroxyl group, either protected or unprotected, the linear arrangement of the hydrophobic moiety, the biphenyl skeleton, and the carboxylic group seem to be essential for the activity of the compounds. PMID- 7473598 TI - Evidence for a carbocation intermediate during conversion of bipinnatin-A and -C into irreversible inhibitors of nicotinic acetylcholine receptors. AB - The lophotoxins are naturally occurring antagonists of nicotinic acetylcholine receptors. These toxins are small diterpenes that irreversibly inhibit nicotinic receptors by specific covalent modification of Tyr190 in the alpha-subunits of the receptor. The naturally occurring lophotoxin analogs, bipinnatin-A and -C, are inactive protoxins. Activation of these toxins occurs spontaneously in buffer and involves replacement of the C2 acetate ester with a hydroxyl group. The mechanism involved in conversion of the inactive bipinnatins into their biologically active solvolysis products was investigated in this study. Solvolysis of bipinnatin-A in buffer containing [18O]water demonstrated that the C2 hydroxyl of the biologically active solvolysis product originated from the solvent. The rates of solvolysis of bipinnatins-A and -C were not affected by sodium azide. However, in the presence of azide, solvent products decreased and new azide-containing products appeared. Thus azide acted as a nucleophile after a rate-limiting step, such as the formation of a carbocation intermediate. The kaz/ks values for bipinnatin-A (2900 M-1) and bipinnatin-C (1450 M-1) suggest that the carbocation intermediates are relatively stable. Compounds capable of spontaneously generating carbocations may represent a novel new class of active site-directed affinity reagents that can be applied to other receptors and enzymes. PMID- 7473599 TI - Synthesis and antimicrobial and toxicological studies of amino acid and peptide derivatives of kanamycin A and netilmicin. AB - Amino acid and peptide derivatives of aminoglycosides have been obtained by substitution of the 1-N or 6'-N amino functions of kanamycin A and netilmicin via the temporary complexation of vicinal and nonvicinal amino and hydroxy functions by copper ion [1-N kanamycin A derivatives: L-Ala (6a), D-Ala (6b), Gly (6c), L Asp (6d), L-Ala-L-Ala (6e). 6'-N kanamycin A derivatives: L-Ala (3a), D-Ala (3b), Gly (3c), L-Ala-L-Ala (3e), L-Leu (3f). 6'-N netilmicin derivatives: L-Ala (9a), D-Ala (9b), Gly (9c), L-Asp (9d), L-Ala-L-Ala (9e)]. Characterization was made by FAB-MS, IR, 1H-NMR, and 13C-NMR. All derivatives were essentially inactive. The nephrotoxic potential of the derivatives obtained in sufficient quantities (3b,e and 9a-e) was assessed by measuring their inhibitory potential toward the activity of lysosomal phospholipase A1 acting on phosphatidylcholine embedded in negatively-charged membranes. One compound, 6'-N-L-Ala-netilmicin (9a), showed a 2-fold decrease of inhibitory potency compared to its parent drug. A conformational analysis revealed that it adopts two equally probable conformations and orientations when interacting with phosphatidylinositol. The first in which the drug lies parallel to the hydrophobic-hydrophilic interface, is similar to that of netilmicin. The second, in which the drug inserts itself in the bilayer across the hydrophilic/hydrophobic interface, is similar to that described for streptomycin, an almost non-nephrotoxic aminoglycoside. PMID- 7473600 TI - Topographical modification of melanotropin peptide analogues with beta methyltryptophan isomers at position 9 leads to differential potencies and prolonged biological activities. AB - We have introduced topographical constraints at the 9 position of a superpotent cyclic alpha-melanotropin analogue, Ac-Nle4-Asp5-His6-DPhe7-Arg8-Trp9-Lys10-NH2, by incorporating a methyl group at the beta-carbon of Trp9. These studies were performed on the Trp side chain pharmacophore to identify the bioactive topography of the indole moiety with melanocortin MC1 receptors. The four beta MeTrp9 isomers, in addition to the stereochemical controls L- and DTrp9, were used to probe differential receptor molecular recognition of the tryptophan moiety in two bioassay systems. Approximately a 460-fold difference in potency was observed between the diastereoisomeric peptides in the frog skin bioassay, with only 33- and 10-fold efficacy differences observed in binding and intracellular cAMP accumulation, respectively, on the human melanocortin receptor, hMC1R. The relative orders of potencies in the frog skin bioassay were 2R,3S > 2S,3S = 2R,3R >> 2S,3R and for the hMC1R were 2S,3S > 2R,3R > 2R,3S >> 2S,3R. Of particular interest is the ability of these topographically constrained ligands to differentially affect prolonged biological activity. The 2R,3R diastereoisomeric peptide possessed superprolonged activity, whereas the 2S,3S peptide lacked any residual activity in the frog skin bioassay. However, on the melanocortin receptor, the 2S,3S diastereoisomeric peptide maintained slow dissociation rates (t1/2 = 7 h), while the other diastereoisomeric peptides possessed dissociation t1/2 rates of ca. 2 h. These data strongly implicate ligand-receptor interactions and kinetics as contributing to the observed prolonged biological activities and clearly illustrate topographical recognition differences between these two peripheral MC1 receptors involved in skin pigmentation. This study also demonstrates that topographical modifications of pharmacophore side chain residues, in addition to identifying preferential side chain orientation, can be a useful strategy for the design of peptides to increase the duration of biological activity, relative to the native ligand. PMID- 7473601 TI - A concerted study using binding measurements, X-ray structural data, and molecular modeling on the stereochemical features responsible for the affinity of 6-arylpyrrolo[2,1-d][1,5]benzothiazepines toward mitochondrial benzodiazepine receptors. AB - The 7-(acyloxy)-6-arylpyrrolo[2,1-d][1,5]benzothiazepine derivatives have been recently proposed as a new class of ligands specific for the mitochondrial benzodiazepine receptor (Fiorini et al. J. Med. Chem. 1994, 37, 1427-1438) (Greco et al. J. Med. Chem. 1994, 37, 4100-4108). In this paper we report the X-ray crystallographic structures of three potent (1-3) and two inactive (4 and 5) previously described benzothiazepines, as well as binding affinity constants for two newly assayed analogs in which the acyloxy side chain was replaced by a methoxy group (6) or removed (7). Structure-affinity relationships and molecular mechanics calculations performed using crystal structures as references have led to a revised 3D pharmacophore model accounting for all the data available up until now. Interestingly, the hypothetical receptor-bound conformations of 1-3 display a considerable degree of similarity with their crystal geometries. Additional calculations have confirmed that the poor affinities of benzothiazepines bearing an aroyloxy group (4 and 5) should be ascribed to the steric and/or electronic features of the side chain aryl moieties rather than to unfavorable conformational properties. PMID- 7473602 TI - Inhibition of lecithin: cholesterol acyltransferase activity in human blood plasma by cigarette smoke extract and reactive aldehydes. AB - Cigarette smoking is a risk factor for atherosclerosis. It is conceivable that reactive chemical components in cigarette smoke may adversely affect reverse cholesterol transport at the level of lecithin: cholesterol acyltransferase (LCAT) and promote atherogenesis. Hence, the effect of cigarette smoke extract (CSE) on the activity of LCAT in human plasma was studied. When incubated with plasma, CSE caused both concentration- and time-dependent losses of LCAT activity. Addition of glutathione, but not ascorbate, to plasma prevented loss of LCAT activity caused by CSE. Incubation of plasma with some reactive aldehydes known to be present in cigarette smoke also inhibited LCAT activity. Among five aldehydes tested, acrolein was the strongest inhibitor of LCAT, with complete enzyme inhibition occurring at 1 mM. Acetaldehyde was the weakest inhibitor of LCAT, with 85% enzyme inhibition at 50 mM. Hexanal, formaldehyde, and malondialdehyde completely inhibited LCAT activity at 10, 50, and 50 mM, respectively. When plasma was incubated with 1 mM acrolein in the presence of 2.5 mM glutathione or dihydrolipoic acid, 100 and 57% of LCAT activity, respectively, remained after incubation. This finding suggest that reactive aldehydes may form adducts with certain free sulfhydryl groups functioning in the active site of LCAT to inhibit enzyme activity. It is concluded that reactive aldehydes are at least partially responsible for the reduction in LCAT activity in plasma treated with CSE. PMID- 7473603 TI - The effects of hexachlorobenzene on circulating levels of adrenal steroids in the ovariectomized rat. AB - Hexachlorobenzene (HCB), is a global pollutant that resists degradation and possesses a propensity to bioaccumulate. However, the effect of HCB on adrenal function remains largely unknown. Thus, circulating levels of adrenal steroids in HCB-exposed (0.0, 1.0, 10.0, or 100.0 mg/kg/day--for 30 days by gavage) adult ovariectomized Sprague-Dawley rats (N = 32) were investigated. A terminal blood sample was collected for HCB residue analysis, and levels of circulating progesterone (P4), corticosterone (CS), and aldosterone (ALD) were quantified. Mean serum CS levels were significantly (P = 0.02) reduced by HCB exposure, starting with the lowest dose group (1.0 mg/kg/day for 30 days), whereas no differences in mean serum P4 and ALD levels were observed. Since it has been argued that the rodent possesses the ability to produce small amounts of cortisol and that levels of this glucocorticoid are altered in pathological states, serum cortisol (C) levels were also measured. Circulating levels of C were significantly lower (P < 0.05) in the highest dose group compared with controls. The presence of C in serum was confirmed by reverse-phase HPLC. These data suggest that even at the lowest dose studied, HCB exposure induces alterations in steroidogenesis of cells of the adrenal cortex inner zone. PMID- 7473604 TI - Nucleotide changes in oxidatively stressed lymphocytes. AB - Similar to HIV-1-induced suppression of thymus-derived lymphocytes (T cells), oxidatively stressed T cells show inhibited DNA synthesis and proliferation. The influence of oxidative stress on nucleotide pools was explored using 3H-uridine addition to OKT3-stimulated peripheral blood lymphocytes. The cells were preincubated and stimulated in the presence of 1 mM buthionine sulfoximine to inhibit GSH synthesis. This treatment gave rise to a significant reduction in dUDP and TTP biosynthesis following 18-32 hours stimulation, indicating possible impairment of ribonucleotide reductase activity. PMID- 7473605 TI - Paradoxical effect of Sudan III on the in vivo and in vitro genotoxicity elicited by 7,12-dimethylbenz(a)anthracene. AB - Effect of the induction of drug metabolizing enzymes by Sudan III on the in vivo and in vitro genotoxicity elicited by 7,12-dimethyl-benz(a)anthracene (DMBA) was investigated. A significant suppression of DMBA-induced micronucleated reticulocytes was observed in C57BL/6 mice treated with Sudan III intraperitoneally for 3 or 5 days before injection of the DMBA. However, the preincubation of DMBA with hepatic microsomes from Sudan III-treated rats caused a marked increase in the in vitro mutagenicity in the Ames assay, paradoxically. Sudan III was found to induce CYP 1A1, 7-ethoxycoumarin O-deethylase activity as well as both UDP-glucuronyl transferase and glutathione S-transferase activities. The increase of mutagenicity of DMBA observed in the Ames assay using hepatic microsomes from Sudan III-treated rats was inhibited by the addition of uridine 5'-diphosphoglucuronic acid or reduced glutathione with cytosol. Mutagenic metabolites of DMBA formed by CYP1A1 appeared to be effectively detoxified by these phase II enzymes. The results of this study suggest that Sudan III-induced prevention of in vivo mutagenesis is due to the induction of both CYP 1A1 and detoxifying phase II enzymes. The induced CYP1A1 may accelerate formation of active metabolic intermediates, but phase II enzymes are also induced and detoxify these intermediates to inactive metabolites. This would reduce residence time of the carcinogen in the body and the time of exposure to active metabolites for target organs. PMID- 7473606 TI - Characterization of the Ah receptor from human placental tissue. AB - The rate of thermal inactivation of the unliganded human Ah receptor, studied by sucrose density gradient centrifugation, with respect to loss of ligand binding ability, was found to be greater than those of most rodents at 20 degrees C, but the temperature coefficient of the rate constant was much smaller than for the rodent species. This implies that the unliganded human Ah receptor would be thermally more stable than the rodent analogs at physiological temperatures. The liganded form of the human Ah receptor was found to be less stable with respect to ligand release than the rodent receptors. These differences in behavior between human and rodent Ah receptors underline the difficulties in using rodent data in the development of receptor-based models of dioxin toxicity. Attempts to develop an alternative to sucrose density gradient centrifugation, comparable with the hydroxylapatite adsorption method used to assay rodent hepatic Ah receptor, were unsuccessful. PMID- 7473607 TI - Alteration of Ca2+ homeostasis of sea urchin embryos by retinoid CD 367, dual effect on egg cleavage and embryonic development. AB - The effect of a light stable retinoid (CD 367) was studied on sea urchin embryos. CD 367 did not affect sperm-egg interaction. In a range of concentrations between 10 and 100 microM, CD 367 delayed the first and the second cleavages. When added after fertilization, micromolar amounts of CD 367 delayed hatching and produced embryonic abnormalities in a dose-dependent manner. Mesodermal cells, primary (PMC) and secondary (SMC) mesenchyme cells migration was particularly disturbed, leading to exogastrulations and calcified spicules malformations. Concentrations of CD 367 higher than 8 microM were embryolethal. Micromolar amount of CD 367 increased plasmalemma Ca2+ permeability of fertilized eggs but not of unfertilized eggs. CD 367 inhibited ATP-dependent intracellular sequestration of Ca2+ in a range of concentrations similar to those affecting egg cleavage and embryonic structures. Since we were unable to detect nuclear receptors for CD 367 in sea urchin eggs and ovocytes, these effects probably are not related to interaction of the retinoid with members of the RAR family, to which CD 367 has a high affinity, but rather to its toxicity by the means of some unknown mechanisms. PMID- 7473608 TI - Gender differences in hepatic expression of flavin-containing monooxygenase isoforms (FMO1, FMO3, and FMO5) in mice. AB - Hepatic flavin-containing monooxygenase (FMO) activity of microsomes from adult CD-1, Swiss-Webster, C57BL/6, and DBA/2 mice was found to be significantly higher in females than in males. Based on protein and mRNA levels in CD-1 mice, FMO forms responsible for the gender difference in FMO activity were FMO1 and FMO3. FMO1 expression was two to three times higher in female mice compared with males; FMO3, however, which was expressed at levels equivalent to FMO1 in female mice, was not detected in males. The expression of FMO5 was approximately equal in both sexes. FMO2 and FMO4 transcripts were not evident in hepatic mRNA from mice. Protein and mRNA levels appear to be coregulated with regard to gender-selective or gender-specific expression of FMO1 or FMO3, respectively. FMO5, which demonstrates no gender-selective expression in mice, may be regulated by different mechanisms. Examination of protein levels among Swiss-Webster, C57BL/6, and DBA/2 strains revealed a gender-dependent expression of FMO isozymes identical to the CD-1 strain. PMID- 7473609 TI - Host-feeding patterns of Culex pipiens and Cx. antennatus (Diptera: Culicidae) from a village in Sharqiya Governorate, Egypt. AB - Host-selection patterns of mosquitoes were determined over a 1-yr period at Abu Heif, a village in Sharqiya Governorate, Egypt, that has had a history of Rift Valley fever virus (RVFV) transmission. Culex pipiens L. and Cx. antennatus Becker were the most common mosquito species collected, and 8,252 blood meals from both species were analyzed by a precipitin technique. Host availability was estimated by a monthly census of human and animal populations. Both mosquito species exhibited opportunistic endophagic behavior. In bedrooms, 79% of Cx. pipiens fed on humans, compared with 53% of the Cx. antennatus. In animal sheds, 35% of Cx. pipiens and 68% of Cx. antennatus fed on sheep or goats. Cx. pipiens was primarily anthropophilic (forage ratio = 2.7) whereas Cx. antennatus was mainly an ovine feeder (forage ratio = 2.4). These findings indicated that both species probably were involved in the transmission of RVFV in Sharqiya Governorate during the epidemics of 1977 and 1978, Cx. pipiens being mainly responsible for transmission to humans, and Cx. antennatus for transmission among domestic animals. The persistent custom of keeping sheep and goats inside human dwellings, combined with the opportunistic host selection by the local mosquitoes, continues to make this area receptive to RVFV transmission. PMID- 7473610 TI - Bacteriocidal qualities of ixodid tick (Acarina: Ixodidae) salivary cement plugs and their changes under the influence of a viral tick-borne pathogen. AB - The abundance of bacteriocidal compounds contained in the salivary cement plug of ixodid ticks was changed because of the reproduction of tick-borne encephalitis virus (TBEV) in their bodies. The size of TBEV-infected Ixodes persulcatus Shulze lytic zone surrounding the cement plug enlarged to that of naive ticks, whereas Micrococcus lysodeikticus (Cohn) lytic zones induced by the cement plugs of TBEV infected Amblyomma hebraeum Koch nymphs or Rhipicephalus appendiculatus Neumann females were reduced, compared with those produced by noninfected specimens. It is possible that an increase of lysozyme production by the primary TBEV vectors (Ixodinae) infected salivary gland cells, compared with suppression of the bacteriocidal qualities of saliva of ticks that are not TBEV vectors in nature (Amblyomminae), is an indication of the specificity of a I. persulcatus-TBEV interface. PMID- 7473611 TI - Laboratory comparison of feeding success among Triatoma infestans, T. guasayana, and T. sordida (Hemiptera: Reduviidae). AB - Success in obtaining a blood meal and rapid access to hiding places after feeding are the principal requirements triatomines have as they colonize artificial ecotopes. Feeding success and postfeeding location of 3rd and 5th instars of Triatoma sordida (Stal), of T. guasayana Wygodzinsky & Abalos, and T. infestans (Klug) were studied in an experimental box in which triatomines and a chicken were left to interact overnight. The bird was enclosed in a glass cube, slightly raised to allow triatomines to get in and out, turning the space outside into an extensive refuge area. Four bunches of dry corn husks and a wooden panel were also offered as shelters. The number of live and dead insects and their locations at the end of the experiment were recorded. Predation--as the percentage of missing insects--and success--as the percentage of insects alive and fed--were calculated. Interactions with the host were different among species and sometimes also between individuals of different life stages of the same species. Predation was always > 20%. T. sordida was the best exploiter of the blood source, because 3rd instars were more successful and 5th instars were as successful as the corresponding T. infestans stages. Performance of T. guasayana was equivocal because survival and feeding success were different for both instars. T. infestans showed a pronounced tendency to climb, and 3rd instars of T. guasayana were equally distributed in the upper half of the box and in the corn husks, whereas the majority of T. sordida and 5th instars of T. guasayana sheltered in the husks. PMID- 7473612 TI - Effects of two insect growth regulators on ecdysteroid production in Aedes aegypti (Diptera: Culicidae). AB - Diflubenzuron and OMS 2017 are insect growth regulators that affect larval to adult development in Aedes aegypti (L.) by altering ecdysis. When larvae were exposed to sublethal concentrations, surviving adults express reduced reproductive potential. In mosquitoes, ecdysteroids are important in larval and adult ovarian development. We applied 30% emergence reduction concentrations (EI30) of OMS 2017 and diflubenzuron to 4th-instar Ae. aegypti to determine if changes in ecdysteroid production may explain these physiological effects. Ecdysteroid concentrations were measured in the larvae, pupae, and adults after treatment with both IGRs. After treatment with OMS 2017, the 1st peak of ecdysteroid production in larvae was totally inhibited, but after pupation, ecdysteroid concentrations were the same as in untreated controls. In diflubenzuron-treated larvae, the second peak of production was delayed and the ecdysteroid concentrations of the larvae, pupae, and adults were depressed slightly when compared with untreated controls. The production of ecdysteroids by the ovaries was not altered by sublethal larval treatment with both IGRs. Although OMS 2017 and diflubenzuron belong to the same chemical family, their mode of action apparently is different. PMID- 7473613 TI - Sylvatic triatomines (Hemiptera: Reduviidae) in Bolivia: trends toward domesticity and possible infection with Trypanosoma cruzi (Kinetoplastida: Trypanosomatidae). AB - The risk of domestic transmission of Trypanosoma cruzi (Chagas) by sylvatic triatomines was assessed in an isolated area of the subandean region of Bolivia. None of the 390 residents examined had serological evidence of infection. Two sylvatic triatomine species, Eratyrus mucronatus (Stal) and Triatoma sordida (Stal), were found in houses and in peridomestic structures. The collection of nymphal instars of both species from some houses indicated possible domesticity. Microscopic examination of feces from 92 insects showed no parasites, and cultures from the guts of 30 insects were negative. Nevertheless, a polymerase chain reaction (PCR) test performed on the same fecal samples showed the presence of T. cruzi DNA in 19.1 and 12.5% of E. mucronatus and T. sordida, respectively. These 16 PCR-positive samples were hybridized with 2 T. cruzi-specific probes known from the domestic cycle in Bolivia (clones 20 and 39). At least 1 of these clones was identified in 7 bugs (5 E. mucronatus and 2 T. sordida). Moreover, no hybridization was observed with these probes in S E. mucronatus and 1 T. sordida samples that showed an amplified band by PCR. These data indicated that T. cruzi clones, genetically unrelated to clones 20 and 39, also were circulating in this area. Based on these results, the 2 sylvatic triatomine species encountered in Apolo should not be overlooked as possible local vectors of T. cruzi. PMID- 7473614 TI - Aedes albopictus (Diptera: Culicidae) eggs: field survivorship during northern Indiana winters. AB - Aedes albopictus (Skuse) is an Asian mosquito that recently has colonized North America via used tire transport. Temperate Ae. albopictus populations overwinter as diapausing eggs, but tropical populations cannot diapause. Eggs of tropical (SABAH) and temperate (INDY) Ae. albopictus were obtained in diapause-inducing conditions and placed inside a scrap tire to monitor overwintering survival in northern Indiana during the winters of 1989-1990 and 1990-1991. Diapause eggs of Ae. triseriatus (Say), a native North American mosquito, were included for comparison. Tropical Ae. albopictus from Malaysia did not survive either winter. Temperate Ae. albopictus from Indianapolis, IN, did not survive the winter of 1989-1990, but 78% survived the winter of 1990-1991. In contrast, 92 and 96% of Ae. triseriatus survived the winters of 1989-1990 and 1990-1991, respectively. Neither mean temperature nor absolute minimum temperature (a winter's lowest temperature) accurately predicted Ae. albopictus overwintering survivorship in the field. The possible effect of snow and other insulating materials on the overwintering survivorship of Ae. albopictus eggs is discussed. PMID- 7473615 TI - Nocturnal activity patterns of Lutzomyia longipalpis (Diptera: Psychodidae) at an endemic focus of visceral leishmaniasis in Colombia. AB - Nocturnal activity of the sand fly Lutzomyia longipalpis (Lutz & Neiva) was studied from August 1991 to July 1992 in a small rural community in Colombia where American visceral leishmaniasis is endemic. During 2 or 3 nights each month, sand flies were collected with hand-held aspirators each hour between 1730 and 0630 hours, from a pigpen and a cattle corral located 30 m apart. Host seeking activity of L. longipalpis adults was characterized by 2 general patterns: (1) adult sand fly activity increased shortly after sunset and continued until just after sunrise, and (2) peak sand fly activity was greatest early in the evening (1830-2330 hours) and then declined steadily toward morning. Female L. longipalpis activity generally increased after 2030 hours, whereas that of males remained constant or declined as the evening progressed. There were seasonal differences in sand fly abundance between the 2 sites: peak abundance in the cattle corral occurred during hot, dry periods, whereas maximum abundance in the pigpen occurred when relative humidity was higher. Influence of relative humidity on activity varied with season. Sand fly activity tended to decrease at temperatures below 24 degrees C and increase in the presence of moonlight. PMID- 7473617 TI - Antibody responses in cattle infested with Haematobia irritans irritans (Diptera: Muscidae). AB - Antibody response to feeding by the horn fly, Haematobia irritans irritans (L.), was measured in naturally infested cattle as well as in controlled infestations. Humoral antibody level was not correlated with fly abundance in natural infestations. Correlations between the antibody response and fly abundance were extremely variable among animals. Antibody response to horn fly salivary antigens peaked within 4 wk after fly abundance reached 150 flies per animal. In controlled infestations, antibody response was weak during an initial exposure to 200 flies per animal, but increased after flies were removed from the cattle. Horn fly salivary antigen may have an immunomodulatory effect on the host. PMID- 7473618 TI - Effect of temperature on Culex tarsalis (Diptera: Culicidae) from the Coachella and San Joaquin Valleys of California. AB - The F1 progeny of Culex tarsalis Coquillett females from the Coachella (CV) and San Joaquin (SJV) valleys collected during April 1991, and April, July, and October 1993 were reared and maintained as adults at 5 constant temperatures ranging from 14 to 38 degrees C. CV F1 progeny exhibited smaller body size, enhanced survival during spring, and higher autogeny rates than SJV F1 progeny; however, upper and lower thermal tolerance limits, immature developmental rates and survivorship, and adult life table parameters were relatively similar for both strains. Mosquitoes from both sites exhibited midsummer changes in immature developmental rates and survivorship, adult wing length, life expectancy at emergence, and generation time. These data indicated that temperature may select for both spatial and temporal changes in mosquito biology. PMID- 7473616 TI - Age structure, blood-feeding behavior, and Leishmania chagasi infection in Lutzomyia longipalpis (Diptera: Psychodidae) at an endemic focus of visceral leishmaniasis in Colombia. AB - Ecological studies on the sand fly Lutzomyia longipalpis (Lutz & Neiva) were conducted during 1990-1992 in a small rural community in Colombia where American visceral leishmaniasis (AVL) is endemic. Subsamples of sand flies collected weekly from pigpens, the interior of houses, and natural outdoor resting sites were dissected to determine physiological age and Leishmania chagasi Cunha & Chagas infection rates. Eleven female L. longipalpis had flagellates in their gut, 2 of which were successfully cultured and identified as Leishmania chagasi. The reproductive status, stage of ovarian development, and trophic history of female sand flies varied among sites, habitats, and time of collection. The percentage of parous females ranged from about one-third to two-thirds overall and varied seasonally. Of most relevance to AVL transmission was the finding that 8% of L. longipalpis females were multiparous. In addition, our data suggest that L. longipalpis rest inside houses after blood-feeding outdoors, and that this species can blood-feed more than once during a single gonotrophic cycle. PMID- 7473620 TI - Prey density effect on cannibalism by Toxorhynchites towadensis (Diptera: Culicidae). AB - The effect of prey density on cannibalism by the larvae of Toxorhynchites towadensis (Matsumura) was studied in the laboratory. Frequency of cannibalism was compared among pairs of all possible instar combinations of Tx. towadensis in the presence and absence of Aedes albopictus (Skuse). Cannibalism occurred despite the presence of prey larvae, but more frequently between different instars than the same instars. Cannibalized larvae were always younger than or the same age as their attackers. Six predatory larvae were reared in a container with 30, 60, or 120 prey throughout their development, and cannibalism and development were observed. No cannibalism was observed when prey density was 120. However, in the continuous presence of 30 prey, cannibalism occurred on 8 occasions. Cannibalism at a prey density of 60 was intermediate in frequency. Most (92%) cannibalism occurred when 1st or 2nd instars attacked the same or younger instars. The time in cohabitation of different instars was shorter with increasing prey density. PMID- 7473619 TI - Effects of seven antibiotics on the growth and development of Phaenicia sericata (Diptera: Calliphoridae) larvae. AB - Maggot therapy is used for patients with severe tissue destruction, who often receive antibiotics concurrently. Therefore, we studied the effects on maggots of ampicillin, cefazolin, ceftizoxime, clindamycin, gentamicin, mezlocillin, and vancomycin in concentrations of 1, 10, 100, and 1,000 times the average minimum bactericidal or bacteriostatic concentration against highly susceptible organisms. There was a reduction in larval survival in media with gentamicin at concentrations of 1,000 times the average pharmacologic level, or 4,000 micrograms/ml (2.7% survival), versus lower concentrations (80-88% survival). Maturation of the surviving pupae to adults also was decreased at this concentration. Media with cefazolin concentrations of 100 times the average bacteristatic level, or 800 micrograms/ml, also led to a significant decrease in larval survival (70% versus 80-88%). There were no differences in larval survival, rate of maturation, or pupal weights for organisms reared on media containing ampicillin, ceftizoxime, clindamycin, mezlocillin, or vancomycin. P. sericata matured normally--and thus can be used therapeutically--when exposed to standard pharmacologic levels of the 7 antibiotics tested. Furthermore, the methods employed herein may be helpful to forensic entomologists attempting to develop models for drug ingestion by maggots. PMID- 7473621 TI - Short-term reproductive diapause by Culex tarsalis (Diptera: Culicidae) in the Coachella Valley of California. AB - Culex tarsalis Coquillett from the Coachella Valley in southeastern California enter a short-term reproductive diapause that is terminated by changes in photoperiod at or shortly after the winter solstice. Reproductively active and unfed resting females exhibited respective increases and decreases in parity rates during December that were characteristic of diapause induction, although reproductively active females, larvae, and males were collected during every month throughout winter. Laboratory experiments demonstrated that females from Coachella Valley enter and maintain a facultative reproductive diapause similar to females from the more northern San Joaquin Valley when exposed to simulated winter conditions. In environmental chambers, diapause termination was not related to temperature accrual because females terminated diapause at approximately the same time regardless of temperature regimens. A field experiment showed that female cohorts emerging in the Coachella Valley from late October through December entered a reproductive diapause that was terminated synchronously by changing photoperiod in late December. PMID- 7473622 TI - Genetic differentiation and diagnostic loci of Anopheles nuneztovari, An. trinkae, and An. rangeli (Diptera: Culicidae). AB - Samples of Anopheles rangeli Gabaldon, Cova Garcia & Lopez, An. trinkae Causey, and An. nuneztovari Gabaldon from Venezuela, Ecuador, Brazil, and Bolivia were analyzed for genetic variability at 24 enzyme loci. Estimates of genetic variability for An. rangeli and An. trinkae from Ecuador and for An. nuneztovari in Venezuela had the following ranges: 46-58% polymorphic loci, 1.7-2.0 (SEM = 0.1-0.3) mean number of alleles per locus, and 0.069-0.113 (SEM = 0.03-0.04) expected mean heterozygosity. Genetic variability estimates of An. rangeli from Bolivia were 20.8-29.2% polymorphic loci, 1.2-1.6 (SEM = 0.1-0.2) mean number of alleles per locus, and 0.037-0.054 (SEM = 0.02-0.03) expected mean heterozygosity. The estimated genetic distance between An. rangeli and An. trinkae ranged from 0.149 to 0.197. The genetic distance between these 2 species and An. nuneztovari ranged from 0.319 to 0.440. Although there were allele frequency differences at some loci between samples of An. nuneztovari sampled from either side of the Andes Mountains in Venezuela, there were no diagnostic loci and the estimated genetic distance was only 0.023. Seven enzyme loci were diagnostic between An. nuneztovari and one or both of its sister species: Acon-2, Ao, Hk-1, Idh-2, Me, Pgi, and Pgm. The diagnostic loci Hk-1 and Acon-2 were not polymorphic in any species. An. rangeli and An. trinkae can be distinguished by the diagnostic loci Ao, Idh-2, and Me-1, and with a 97% probability by Pgm. Distance Wagner and unweighted pair-group method with arithmetic averaging analyses support a close phylogenetic relationship between An. trinkae and An. rangeli. PMID- 7473623 TI - Pterin quantity and gonotrophic stage as indicators of age in Musca autumnalis (Diptera: Muscidae). AB - Spectrofluorimetric methods were used to measure pterin quantities in the head capsules of adult face flies, Musca autumnalis De Geer. Quantities at eclosion were greater in males than in females, and were proportional to the head capsule width in each sex. Pterin accumulated as a linear function of time at constant temperature, at rates of 3.8 relative fluorescence units per degree-day in males and 1.68 units in females, both above a common base of 9.8 degrees C. A mark release-recapture experiment revealed that pterin accumulated at a rate of 6.40 units per degree-day among males, twice as fast as in the laboratory. In contrast, accumulation among females was 1.35 units per degree-day, approximating their laboratory rate. Gonotrophic age grading of recaptured females indicated median development rates corresponded with expectations based on earlier laboratory studies, but substantial variation among females led to imprecise estimates of chronological age. Calibration curves for each sex are presented to estimate degree-day age from head capsule width and pterin content. The pterin age-grading method allows the field study of longevity of male face flies, and complements gonotrophic methods for the study of longevity and reproductive success of females. PMID- 7473624 TI - Age structure and reproductive composition of summer Musca autumnalis (Diptera: Muscidae) populations estimated by pterin concentrations. AB - Pterins in face flies, Musca autumnalis DeGeer, were measured spectrofluorimetrically to estimate the chronological ages of adults in natural summer populations. The age distributions showed a systematic undersampling of young males. Exponential, Gompertz, and Weibull models were fitted to the age distributions. A 1-parameter exponential model suggested mean daily survival rates of 0.83 for female face flies, and a mean life expectation of 5.5 d. But the Weibull model best fit the age distributions, providing mean life expectations of 10.1 and 11 d for males and females, respectively. Ovarian dynamics were examined with respect to chronological age and delays were detected in vitellogenesis and oviposition. Face fly populations seemed to reproduce at less than the maximum rate allowed by prevailing temperatures, at which it was estimated that an average female would undergo 2.2 ovarian cycles and have a lifetime fecundity of 28.5 female progeny. PMID- 7473625 TI - Diagnosis by random amplified polymorphic DNA polymerase chain reaction of four cryptic species related to Anopheles (Nyssorhynchus) albitarsis (Diptera: Culicidae) from Paraguay, Argentina, and Brazil. AB - Random amplified polymorphic DNA polymerase chain reaction (RAPD-PCR) analysis was applied to samples from widespread populations of the poorly characterized Anopheles (Nyssorhynchus) albitarsis Lynch-Arribalzaga species complex, and 4 genetically differentiated species were distinguished. A screen of 65 random decamer oligonucleotide primers identified 12 primers, which produced 19 reproducible species-specific genetic markers and 4 markers common to 2 or more species. These markers were correlated in nearly all individuals of each species throughout the ranges sampled, including populations as far apart as 2,500 km. Each individual analyzed was from a different isofemale progeny brood, with associated morphological specimens. These specimens will facilitate studies to relate these species to previously reported chromosomal and enzymatic variation as well as to their feeding behavior and potential as malaria vectors. We hypothesize that 3 of the species have recognized valid names: An. (Nys.) albitarsis Lynch-Arribalzaga, An. (Nys.) marajoara Galvao and Damasceno, and An. (Nys.) deaneorum Rosa-Freitas, whereas the 4th is undescribed. PMID- 7473626 TI - Field evidence for multiple host contacts during blood feeding by Culex tarsalis, Cx. restuans, and Cx. nigripalpus (Diptera: Culicidae). AB - Field studies were conducted in Manitoba, Canada, and Florida, United States to evaluate the frequency of multiple host contacts by Culex tarsalis Coquillett, Cx. restuans Theobald, and Cx. nigripalpus Theobald, primary and secondary vectors of several encephalitis viruses in North America. Blood-fed mosquitoes were collected from box traps, each baited with a pair of quail. One quail of each pair was injected with rubidium and the other with cesium to permit the determination of the source(s) of each blood meal. Approximately 5% of all blood fed Cx. tarsalis, Cx. restuans, and Cx. nigripalpus ingested blood from both quail during overnight exposure. The frequency of multiple feeding by these species ranged from 0 to 18.5%, 0 to 33.3%, and 0 to 17.6%, respectively. Of the 331 mosquitoes that ingested blood from 2 quail, 57 (17.2%) were not fully blood fed. Also, 1.0-3.5% of blood-fed mosquitoes had ingested blood before entering the quail-baited traps. Any increase in the number of host contacts as a result of multiple feeding, however modest, may increase disproportionately the rate at which virus transmission occurs. PMID- 7473627 TI - Peritrophic envelopes of Lutzomyia spinicrassa (Diptera: Psychodidae). AB - The secretion, morphology, and chemical composition of the peritrophic envelope were studied in the phlebotomine sand fly, Lutzomyia spinicrassa Morales, OsornoMesa, Osorno & Hoyos, a suspected vector of Leishmania braziliensis in Colombia and Venezuela. Viewed under light microscopy, the envelope matured rapidly and could be dissected from the blood bolus as early as 12 h and until 36 h after feeding; subsequently it began to degrade. The envelope was initially a closed sac around the blood meal, but opened posteriorly in most flies by 6 h. The posterior opening may facilitate the migration and establishment of Le. braziliensis in the hindgut. Secretion of envelope precursors was from the entire midgut epithelium. Electron microscopy revealed that electron-dense precursor material (possibly chitin) was present, bathing the microvilli during the first 12 h after blood feeding. This secretion appeared to originate from the bases of the microvilli. From 1 to 36 h, an electron-lucid precursor material (possibly protein) was secreted from the entire length of microvilli and from their bases. Both precursors appeared to be formed at the epithelial surface, not associated with secretory vesicles. The envelope developed rapidly from precursor material, and by 6 h a defined electron-lucid structure was present above the microvilli. Most mature envelopes (12-36 h) were 0.5-2.1 microns thick, multilayered, wholly electron-lucid, and composed of microfibrils and granules. Electron-dense components were seen in some envelopes at 24-36 h. An anterior hyaline plug was present from 12 to 36 h. Envelopes were composed of chitin, protein, and glycoprotein, based on chemical and histochemical tests. The likely presence of several amino acids (lysine, aspartic acid, and glutamic acid) that may cross link chitin and protein was demonstrated by a positive ninhydrin-Schiff test. This study constitutes the first ultrastructural investigation of peritrophic envelope development by a New World sand fly. PMID- 7473628 TI - Status of Haematobia irritans (Diptera: Muscidae) insecticide resistance in northeastern Mexico. AB - Horn fly, Haematobia irritans (L.), populations in northeastern Mexico were surveyed for resistance to fenvalerate and coumaphos. Compared with a susceptible strain of horn flies, resistance levels among field populations for the pyrethroid, fenvalerate, ranged from 36 to 199 times at the LD50 level, indicating that horn fly control with pyrethroids would be unsatisfactory. Resistance to the organophosphate compound, coumaphos, was less evident, with flies at only 1 of 5 locations showing levels of tolerance that might interfere with adequate control. PMID- 7473629 TI - Suppression of subadult Ixodes scapularis (Acari: Ixodidae) following removal of leaf litter. AB - Removal of leaf litter in wooded areas of a forested residential community significantly reduced the abundance of questing blacklegged tick, Ixodes scapularis Say, nymphs throughout their peak activity period. Removal of leaf litter, using hand rakes and leaf blowers during the early spring (March) and early summer (June) resulted in reductions in nymphal tick density ranging from 72.7 to 100%. Subsequent sampling of the I. scapularis population during the summer revealed similar rates of suppression of larvae. Although leaf removal provides levels of suppression similar to more traditional methods, including chemical acaricides, it is labor intensive and not free of possible nontarget effects. The potential role of leaf litter removal in an integrated program to control I. scapularis is discussed. PMID- 7473630 TI - Dirofilaria (Filarioidea: Onchocercidae) infection in Aedes albopictus (Diptera: Culicidae) collected in Louisiana. AB - First-stage Dirofilaria larvae were found infecting the Malpighian tubules in 3 of 163 Aedes albopictus collected from New Orleans, LA, during June-July 1994. Further studies will be necessary to provide definitive identification of the parasite species, and to better describe the vectorial status of Ae. albopictus. PMID- 7473631 TI - Incompetence of white-tailed deer as amplifying hosts of vesicular stomatitis virus for Lutzomyia shannoni (Diptera: Psychodidae). AB - Sand flies, Lutzomyia shannoni Dyar, were allowed to feed on 3 white-tailed deer, Odocoileus virginiana Zimmermann, that previously had been infected with the New Jersey serotype of vesicular stomatitis (VSNJ) virus. Flies fed in the lower abdominal area of each deer on days 1-5 postinfection. A blood sample, nasal swab, and throat swab were taken during each feeding trial and examined for virus. Blood-fed flies were held for 4-5 d following the bloodmeal and tested for VSNJ virus infection. VSNJ virus was never detected in blood or from swabs taken from infected deer nor from any of the sand flies that fed on deer. The findings suggest that white-tailed deer do not fulfill the traditional concept of amplifying hosts of VSNJ virus. PMID- 7473632 TI - Incompetence of domestic pigs as amplifying hosts of vesicular stomatitis virus for Lutzomyia shannoni (Diptera: Psychodidae). AB - Seven domestic pigs, Sus scrofa L., were infected by intradermal inoculation at 3 different sites with the New Jersey serotype of vesicular stomatitis (VSNJ) virus. Laboratory-reared Lutzomyia shannoni Dyar sand flies, a suspected biological vector of VSNJ virus, were allowed to feed on pigs at the lower abdomen or at sites of their own selection on days 1-7 and on day 10 postinfection. Blood samples were taken from infected swine concomitant with most feeding trials and tested for the presence of virus. Sand flies were held for up to 5 d following ingestion of blood and tested for VSNJ virus infection. Virus was not recovered from the blood of infected pigs or from any of the flies that fed on these pigs. The findings suggest that domestic pigs do not fulfill the traditional concept of amplifying hosts of VSNJ virus. PMID- 7473634 TI - Occurrence of Lutzomyia anthophora (Diptera: Psychodidae) in Arizona. AB - Males and females of Lutzomyia anthophora Addis were collected by vacuum aspiration from woodrat, Neotoma albigula Hartley, and rock squirrel, Citellus variegatus Bailey, nests along Arivaca Creek in Pima County, Arizona. Additional flies were collected from the same location using CDC miniature light traps supplemented with CO2. These collections extend the recorded geographic distribution of this vector of Leishmania mexicana Biagi westward by approximately 724 km and place Lu. anthophora in a Sonoran desert habitat. PMID- 7473633 TI - Human furuncular myiasis caused by Hermetia illucens (Diptera: Stratiomyidae). AB - Cutaneous myiasis caused by Hermetia illucens (L.) has not been reported previously. We present a case of facultative furuncular myiasis characterized by infestation with a single larva in a woman from Seattle, WA, who had traveled to East Africa. PMID- 7473635 TI - On giving preference to prior volunteers when allocating organs for transplantation. PMID- 7473636 TI - What can medical ethics learn from history? PMID- 7473637 TI - Join the club: a modest proposal to increase availability of donor organs. AB - The shortage of suitable donor organs is the most significant single limiting factor in transplant programmes. More lives could be saved or immeasurably improved if more organs were available. I look at two traditional solutions to the shortfall, and suggest that they are ineffective and/or offensive, and consider the features common to any answer to the problem. I then suggest a third solution: that admission to future transplant lists be conditional on registration as a potential organ donor, outlining its benefits, and defending it against one possible objection. PMID- 7473638 TI - The proof of the vegetable: a commentary on medical futility. AB - Patients with 'persistent vegetative state' (PVS) are often cited in the discussions of ethicists as examples of human beings who are unconscious and do not experience life, and a number of theoretical and practical recommendations have been made on that basis. This article examines the evidence and the theoretical rationale for the belief that people with PVS are unconscious and finds them wanting. This conclusion is related to the practice of the discipline of ethics. PMID- 7473639 TI - Smokers, virgins, equity and health care costs. PMID- 7473640 TI - The annual reports of Local Research Ethics Committees. AB - Each Local Research Ethics Committee (LREC) is expected to produce an annual report for its establishing authority. Reports from 145 LRECs were examined with regard to (a) whether the committees were working within the terms of the most recent guidelines from the Department of Health and (b) observations on the role of LRECs with particular reference to accountability. Most LRECs had produced a report, although their length varied greatly. Most reports showed how seriously the committee took its task. Most committees met many of the guidelines; for example, almost all had two or more lay-members. The guideline most frequently not met was that committees should have no more than 12 members. Many committees review very large numbers of projects (maximum 351). Approximately two-thirds provide details in the annual report of individual project titles, their author and the committee decision; all reports should contain this information. Although it may in fact happen more generally, only 23 per cent of the reports referred to any form of monitoring of the eventual outcome of the research. A significant issue to arise from the reports is the extent to which the framework for the operation of LRECs has been confused by the development of the purchaser-provider split. The paper concludes with suggestions for remedying the situation. PMID- 7473641 TI - The elimination of morality. AB - Anne Maclean has written a book which purports to identify and then definitively to reject the claims which she perceives to be the claims of bioethics. If she is right, then the enterprise to which this journal is dedicated is misconceived and worthless. In this paper, I attempt to show why so far from being not right she is comprehensively wrong, both in her understanding of the nature of bioethics and in the specific moral claims she makes about those she terms 'bioethicists'. Since much of her book-length study is devoted to a criticism of my own work, this paper analyses the extent to which Maclean's criticisms of me, and by extension, of other bioethicists, are sustainable. PMID- 7473642 TI - Aftercare for participants in clinical research: ethical considerations in an asthma drug trial. AB - The issue of aftercare for participants in clinical research was explored in the context of an asthma drug trial. Although there may be financial constraints and practical difficulties with implementation, the results show that it may be feasible for clinical investigators and commercial sponsors to take on some limited responsibility for the medical care of research subjects after clinical trials. However, the ethical implications for this practice remain unclear. On the one hand, society may have a moral obligation to compensate and reward some of its members who assume the risk of research subjects for the benefit of society as a whole. On the other hand, the promise of aftercare may provide an inducement to volunteers which, under certain conditions may be considered morally wrong and scientifically unsound. PMID- 7473643 TI - Ethics-committee authorization in Germany. AB - On 9 August 1994 the German legislature revised the German Drug Law (AMG). Included in the revision is a passage requiring, for the first time, that the sponsors and investigators of clinical studies involving human subjects first obtain the approval of an ethics committee before carrying out such studies. According to the legislation, which takes effect on 17 August 1995, approval is to come from 'an independent ethics committee, set up and administered according to state law [emphasis added]' (1). Although it is clear according to the text that the 16 federal states have been empowered to establish ethics committees within their jurisdictions, this does not mean that the state governments are free to transfer exclusive authority in the matter to their respective medical associations, a step that would effectively abolish Germany's private ethics committees. First, the legislation does not rule out the authorization of private ethics committees. Second, as legal scholars attest, the exclusive control of ethics committees by the medical associations would constitute an illegal monopoly. Third, it is arguable that medical-association ethics committees fail to meet the one prior federal requirement, that of independence. There is a great deal of confusion in Germany today about which kinds of ethics committees (public and/or private) the states will sanction before 17 August 1995. In an attempt to sort things out we present a brief explanation of how ther came to be two kinds of ethics committees in Germany, review the legal battle between the two over the issue of authorization, point out how the German legislature, in passing the recent bill, has missed an opportunity to clarify the issue and, finally suggest why the administration of ethics committees by the medical associations may be incompatible with the requirement that ethics committees be independent. PMID- 7473644 TI - Teaching ethics in psychiatry: a one-day workshop for clinical students. AB - In this paper we describe the objectives of teaching medical ethics to undergraduates and the teaching methods used. We describe a workshop used in the University of Liverpool Department of Psychiatry, designed to enhance ethical sensitivity in psychiatry. The workshop reviews significant historical and current errors in the ethical practice of psychiatry and doctors' defence mechanisms against accepting responsibility for deficiencies in ethical practice. The workshop explores the student doctors' own group ethos in response to ethical dilemmas, and demonstrates how the individual contributes to and is responsible for the group ethos through participation and also through nonparticipation. The student feedback about the workshop is reviewed. The Toronto Ethical Sensitivity Instrument was used to assess whether or not the workshop altered sensitivity. Compared to a control group the attenders' sensitivity was significantly increased (on Student's t-test p equals or is less than 0.002). PMID- 7473645 TI - Chinese Confucian culture and the medical ethical tradition. AB - The Confucian culture, rich in its contents and great in its significance, exerted on the thinking, culture and political life of ancient China immense influences, unparalleled by any other school of thought or culture. Confucian theories on morality and ethics, with 'goodness' as the core and 'rites' as the norm, served as the 'key notes' of the traditional medical ethics of China. The viewpoints of Confucianism on benevolence and material interests, on good and evil, on kindheartedness, and on character cultivation were all inherited by the medical workers and thus became prominent in Chinese traditional medical ethics. Hence, it is clear that the medical profession and Confucianism have long shared common goals in terms of ethics. Influenced by the excellent Confucian thinking and culture, a rather highly-developed system of Chinese traditional medical ethics emerged with a well-defined basic content, and the system has been followed and amended by medical professionals of all generations throughout Chinese history. This system, just to mention briefly, contains concepts such as the need: to attach great importance to the value of life; to do one's best to rescue the dying and to heal the wounded; to show concern to those who suffer from diseases; to practise medicine with honesty; to study medical skills painstakingly; to oppose a careless style of work; to comfort oneself in a dignified manner; to respect local customs and to be polite; to treat patients, noble or humble, equally, and to respect the academic achievements of others, etc. Of course, at the same time, Confucian culture has its own historical and class limitations, which exerted negative influences on traditional medical ethics. Now, if we are to keep up with the development of modern medicine, a serious topic must be addressed. That is how to retain the essence of our traditional medical ethics so as to maintain historic continuity and yet, at the same time, add on the new contents of medical ethics so as to incorporate modern features into our system. Therefore, when trying to reform medical practice in China, we are faced with the urgent need to inherit and promulgate the essence of Confucian ethics--discarding its obsolete concepts--while at the same time building up the new medical ethics that can meet the requirements of the outside world and the future. PMID- 7473646 TI - The association of combined alpha and beta fibrinogen genotype on plasma fibrinogen levels in smokers and non-smokers. AB - OBJECTIVE: To determine in healthy men: (1) whether an extended genotype of the fibrinogen gene cluster using the G/A-455 and the BclI polymorphism of the beta fibrinogen gene and TaqI of the alpha fibrinogen gene explains a significantly larger proportion of variance in plasma fibrinogen levels in either smokers or non-smokers than a single polymorphism (G/A-455); (2) whether there is any evidence for genotype-smoking interaction in the determination of fibrinogen levels. DESIGN: A cross sectional study of healthy, white men recruited at the screening for entry into the Thrombosis Prevention Trial. SETTING: The subjects were drawn from four general practices in the United Kingdom. RESULTS: The frequency of the rare alleles in the sample was 0.19 for the G/A-455 polymorphism (A-455), 0.15 for BclI (B+), and 0.27 for TaqI (T+) alleles. BMI and age made significant contributions to the variance in plasma fibrinogen levels only in non smokers of 5.4% and 2.3% respectively and, in the group as a whole, smoking accounted for 6.6% of the variance. In the non-smokers, of the individual polymorphisms only the G/A-455 showed a significant association with plasma fibrinogen levels (p = 0.03). The mean fibrinogen in non-smokers homozygous for the G-455 allele was 2.54 g/l v 2.85 g/l in those homozygous for the A-455 allele, with the polymorphism explaining 3.6% of the variance in plasma fibrinogen levels in this group. On investigation of the association of fibrinogen levels with combined genotypes, the most significant effect was seen with the combination of the G/A-455 and TaqI polymorphisms, with those with no "fibrinogen raising alleles" having a mean fibrinogen of 2.57 g/l v 3.10 g/l for those with four "fibrinogen raising alleles" (p = 0.0036), and this combination explained 8.9% of the variance in plasma fibrinogen levels (p < 0.005). Although the contribution to variance was greater with the G/A-455/TaqI combination than the G/A-455 polymorphism alone (8.9% v 3.6%), this did not reach significance (p = 0.09). By contrast, in the smoking group, the only significant contribution to the difference in plasma fibrinogen levels was the G/A-455 genotype alone which, after adjustment for BMI and age, contributed 3.8% to the variance (p < 0.05). No interaction was shown between smoking and genotype. CONCLUSION: These data suggest that in non-smokers an extended genotype using the G/A-455 beta fibrinogen gene polymorphism and the TaqI alpha fibrinogen gene polymorphism explains a larger proportion of the variance in plasma fibrinogen levels than any one polymorphism alone, but that smoking has an overriding effect so that other variables such as age and BMI make little additional contribution. PMID- 7473647 TI - Spinocerebellar ataxia 1 (SCA1) in the Japanese in Hokkaido may derive from a single common ancestry. AB - Spinocerebellar ataxia 1 (SCA1) is caused by expansion of an unstable CAG triplet repeat located on the short arm of chromosome 6. Precise mapping has shown a positional relationship to closely linked markers in the order of D6S109-D6S274 D6S288-SCA1-AM10GA-D6S89+ ++-EDN1 from centromere to telomere. The haplotype which cosegregated with the disease was determined in 12 Japanese pedigrees with SCA1. Although the alleles of the SCA1 haplotype varied from pedigree to pedigree, depending on the distance from the SCA1 locus, the affected and presymptomatic subjects carried the same alleles at D6S288 and D6S274. All the families with SCA1 had migrated from either Miyagi or Yamagata Prefectures, neighbouring areas in the Tohoku District, the northern part of Honshu which is the main island of Japan. It seems highly likely that SCA1 in the Japanese, at least those residing in Hokkaido, derives from a single common ancestry. PMID- 7473648 TI - Comparison of CTG repeat length expansion and clinical progression of myotonic dystrophy over a five year period. AB - Myotonic dystrophy (DM) is associated with an underlying CTG trinucleotide repeat expansion at a locus on chromosome 19q13.3. We have determined the repeat length in 23 DM patients with varying clinical severity of symptoms and various sizes of repeat amplification. We confirm that as in previous studies there is no strong correlation between repeat length and clinical symptoms but find that the repeat length in peripheral blood cells of patients increases over a time span of five years indicating continuing mitotic instability of the repeat throughout life. Repeat length progression does not appear to be indicative of clinical progression but age probably is. The degree of expansion correlates with the initial repeat size and 50% of the patients with continuing expansions showed clinical progression of their disease symptoms over the five year study period. PMID- 7473649 TI - Attitudes towards Down's syndrome: follow up of a cohort of 280 cases. AB - A follow up study of a cohort of 280 Down's syndrome (DS) fetuses was initiated in order to estimate the percentage of terminations of pregnancy, the prevalence at birth, the survival of DS children, the attitude of the parents at birth, and the medical and surgical care provided. The present study is a preliminary analysis of the data collected up to the age of 1 year. It shows that 43% of the 280 DS fetuses had died by the end of the first year of life, owing to termination of pregnancy (27%), late spontaneous abortion or stillbirth (4%), or death during the first year of life (12%). Among the 33 children who died, 12 had a common atrioventricular canal, six had another major malformation, three died from infection, one from respiratory distress, two were cot deaths, and one was an infanticide, but eight deaths were unexplained, occurring in children with no known malformation or disease. Among the 185 children still alive after 2 days, 23 (12%) were available for adoption, their mothers having elected to remain anonymous. These results show that in some instances parents or professionals feel justified on one hand in not providing DS children with the necessary care and on the other hand to transfer their responsibilities to the public health system. The influence of prenatal diagnosis of chromosome disorders as a determining factor of the social acceptance of DS is still questionable. PMID- 7473650 TI - Neuropsychological characteristics of Huntington's disease carriers: a double blind study. AB - A cohort of 33 people at risk for Huntington's disease (HD), applying for genetic testing, were tested with a battery of neuropsychological tests covering attentional, visuospatial, learning, memory, and planning functions. A psychiatric rating scale, SCL-90R, was also applied, mainly as a control, since cognitive dysfunction could be ascribed to functional disorders as well as neurodegenerative processes. Self-rating did not indicate any psychiatric symptoms in carriers or non-carriers. However, significantly inferior cognitive functioning in the gene carriers was disclosed by the neuropsychological tests. Primarily, attentional, learning, and planning functions were affected. It is concluded that premorbid cognitive decline occurs in HD. PMID- 7473651 TI - Stringent delineation of Pallister-Hall syndrome in two long surviving patients: importance of radiological anomalies of the hands. AB - We report two unrelated, long surviving patients (2 and 17 years) with syndromal hypothalamic hamartoblastoma. Both showed mild facial dysmorphism (downward slanted palpebral fissures, ptosis, microretrognathia), cleft epiglottis, and developmental delay. The younger child had stenosis of the pulmonary arteries, complex urogenital malformations, and anal atresia. In the oldest patient, the hamartoma caused precocious puberty of the central type, combined with complete hGH deficiency. Both patients showed bony anomalies of the extremities: variable proximal synostosis between central (2nd to 4th) metacarpals or intercalary polydactyly with generalised brachydactyly, severe brachytelephalangism, syndactyly, and nail hypoplasia. Together with the absence of anomalies of cholesterol metabolism, a combination of oral frenula, laryngeal malformations, digestive abnormalities, intercalary polysyndactyly, generalised brachytelephalangism, and nail hypoplasia should allow the delineation of Pallister-Hall syndrome, even when a CNS tumour is absent. The radiological abnormalities are helpful in differentiating Pallister-Hall syndrome from the other syndromes in which hypothalamic hamartoblastoma is observed. This is of major importance for genetic counselling, since Pallister-Hall syndrome may be a dominantly inherited disorder, thus contrasting with most of the other disorders with the CAVE phenotype, which are recessively inherited. PMID- 7473652 TI - Velocardiofacial syndrome: learning difficulties and intervention. AB - Velocardiofacial syndrome, delineated in 1978 by Shprintzen, is a multiple genetic disorder, characterised primarily by cleft palate, cardiovascular anomalies, typical facies, and learning disorders. This syndrome with its recurrent pattern of anomalies has significant implications for education as it is linked to a variety of educationally relevant problems including learning difficulties, behaviour disorders, and speech and language deficits. While clinical research advances and new findings in cytogenetics enable early diagnosis and genetic counselling, ramifications of the syndrome in the field of education are still unknown. Learning difficulties should not be viewed solely as a pathological condition and with data on success of intervention, parents and teachers can be apprised of the information. This paper discusses academic characteristics and the effects of intervention with emphasis on interactive computer based instruction on the development of reading, language, spelling, and numeracy skills. Positive effects of computer based instruction on students' self esteem, motivation, and competence in computer operational skills were also observed and noted. We are encouraged by the enthusiasms of the students for the system, the development of interest in reading, and the transfer of remedial instruction to classroom performance. Comparison of pretest and post-test results indicated significant improvement in reading ability as measured using Neale Analysis. This improvement is in accordance with results obtained using computer testing of specific subskills in reading/language and mathematics. PMID- 7473653 TI - Chromosome 1p terminal deletion: report of new findings and confirmation of two characteristic phenotypes. AB - We report three unrelated patients with small terminal deletions involving 1p36.22-->pter that occurred de novo and compare our patients to the 10 previously reported cases. Although our patients have an identical cytogenetic deletion, patients 1 and 2 share similar clinical features that differ substantially from patient 3. Our patients confirm the existence of two characteristic phenotypes in 1p36.22-->pter deletion. Both phenotypes share some dysmorphic features, but are differentiated by characteristics of growth failure versus macrosomia. In addition, we report the new finding of cardiomyopathy and hydrocephalus in the phenotype associated with growth failure. It is possible that different phenotypic subgroups may exist because of differences in the parental origins of the deleted chromosome or of variations in undetectable amounts of genetic material. PMID- 7473654 TI - Consanguinity among the Saudi Arabian population. AB - This study was conducted on 3212 Saudi families to investigate the prevalence of consanguineous marriages. The families were interviewed and the information on the relationship between the husband and wife was obtained. The overall rate of consanguinity shows that 57.7% of the families screened were consanguineous. The most frequent were first cousin marriages (28.4%) followed by distant relative marriages (15.2%) and second cousin marriages (14.6%). The families were grouped according to the province of their origin and the consanguinity rates were calculated accordingly. There were slight differences in the consanguinity rates in the five provinces, which ranged from 52.1% to 67.7%. In each province first cousin marriages were the most frequently encountered pattern, ranging from 17.9% to 40.9%. The inbreeding coefficient (F) was calculated for each province and ranged from 0.020 to 0.030. Within each province, there were several significant differences among the populations in the different areas. The highest rate of consanguinity was 80.6% in Samtah and the lowest rate was around 34% in Abha in the South Western province. These results place Saudi Arabia among the countries of the world with a high rate of consanguinity. The possible consequences of increased consanguinity are presented and discussed. PMID- 7473655 TI - Congenital anomalies and genetic disorders in families of children with central nervous system tumours. AB - Medical genetic histories of 165 children with primary central nervous system (CNS) tumours and 4599 relatives of these probands were examined to identify birth defects or genetic disorders that may be associated with the aetiology of CNS tumours. Twelve primary malignancies were found in 329 (4%) of the parents of probands. Two of 99 half sibs but no full sibs had malignancies. Twenty-four percent of families had histories warranting consultation for an inherited disorder or birth defect. Single instances of malformations or genetic disorders were reported in 36 families and several disorders were reported in more than one family, including familial hypercholesterolaemia (4), olivopontocerebellar atrophy (2), and familial abdominal aortic aneurysm (2). Although recurring abnormalities were not identified in probands, it is possible that one or more of the birth defects or genetic disorders observed in probands or relatives may be associated with CNS tumourigenesis. PMID- 7473656 TI - Congenital heart disease in spondylothoracic dysostosis: two familial cases. AB - Two familial cases of spondylothoracic dysostosis are reported. Both cases had severe congenital heart disease in addition to the skeletal malformations which are characteristic of the condition. PMID- 7473657 TI - Developmental delay and dysmorphic features associated with a previously undescribed deletion on chromosome 1. AB - We report an 18 month old girl with developmental delay, dysmorphic features, and a karyotype 46,XX,del (1) (p32.1p32.3). To our knowledge the clinical features associated with this deletion have not been reported previously. PMID- 7473658 TI - Anonychia and absence/hypoplasia of distal phalanges (Cooks syndrome): report of a second family. AB - We describe a second family with four affected members in three successive generations with a rare nail dysplasia syndrome. The syndrome, which was first reported by Cooks et al in 1985, is characterised by bilateral nail hypoplasia of digits 1-3, with absence of nails of digits 4-5 of the hands, and total absence of all toe nails. In addition, there is absence/hypoplasia of the distal phalanges of the hands and feet. Our family confirms this syndrome as a distinct entity with autosomal dominant inheritance. PMID- 7473659 TI - Apolipoprotein E epsilon 4 allele is a risk factor for familial and sporadic presenile Alzheimer's disease in both homozygote and heterozygote carriers. AB - While apoliprotein E (ApoE) epsilon 4 allele is now a well established risk factor for familial and sporadic senile Alzheimer's disease (AD), its role in the development of the rarer presenile or early onset type is controversial. Early studies showed no association; later ones found enrichment for the epsilon 4 allele in familial or sporadic types or both. We have ApoE genotyped a series of Scottish people (n = 85) with early onset AD. We find highly significant enrichment for both homozygote and heterozygote ApoE epsilon 4 allele carriers in familial and sporadic early onset AD with a pattern closely resembling that in late onset AD. PMID- 7473660 TI - Direct molecular analysis of myotonic dystrophy in the German population: important considerations in genetic counselling. AB - Myotonic dystrophy (DM) is associated with the expansion and instability of a trinucleotide (CTG) repeat at the DM locus on chromosome 19. Direct genomic analysis in the German population was carried out on 18 DM families, six families with equivocal diagnosis, 69 subjects with equivocal clinical diagnosis, and 100 controls using the polymerase chain reaction (PCR) and a refined Southern protocol. In the majority of the cases molecular analysis confirmed the clinical diagnosis. These included seven cases of congenital DM (CDM) with widely differing gene expansions and instabilities. In most DM families the expanded fragment became larger in successive generations, but we also identified four families with contractions and two families that showed stability of the enlarged fragment during transmission. In four clinically defined DM patients we were unable to detect enlarged CTG repeats. Sequencing of each exon of the DM gene in two of these patients failed to show any mutations. Our cases have important implications for genetic counselling of DM families, highlighting both the diagnostic value of direct genomic analysis and its limitations. PMID- 7473661 TI - Double non-disjunction in maternal meiosis II giving rise to a fetus with 48,XXX,+21. AB - We describe a prenatally detected case of double trisomy involving chromosome 21 and the X chromosome (48,XXX,+21) along with determination of the segregation errors responsible for the double aneuploidy. The patient was ascertained as a result of an abnormal maternal serum analyte screen showing an increased risk for fetal Down's syndrome. Following determination of the abnormal karyotype, pregnancy termination was elected. Microsatellite polymorphisms and cytogenetic heteromorphisms were used to determine that both aneuploidies arose as a result of non-disjunction in maternal meiosis II. These results support hypotheses that a segregation defect at a cellular level may cause non-disjunction involving more than one chromosome. PMID- 7473662 TI - Point mutation of the mitochondrial tRNA(Leu) gene (A 3243 G) in maternally inherited hypertrophic cardiomyopathy, diabetes mellitus, renal failure, and sensorineural deafness. AB - The A 3243 G mutation of the mitochondrial tRNA(Leu) gene was found to segregate with maternally inherited diabetes mellitus, sensorineural deafness, hypertrophic cardiomyopathy, or renal failure in a large pedigree of 35 affected members in four generations. Presenting symptoms almost consistently involved deafness and recurrent attacks of migraine-like headaches, but the clinical course of the disease varied within and across generations. The A 3243 G mutation has been previously reported in association with the mitochondrial encephalomyopathy, lactic acidosis, and stroke-like episode syndrome (MELAS) and with diabetes mellitus and deafness. To our knowledge, however, hypertrophic cardiomyopathy is not a common feature in people with the A 3243 G mutation and renal failure has not been hitherto reported in association with this mutation. The present observation gives additional support to the variable clinical expression of mtDNA mutations in humans. PMID- 7473663 TI - Recurrence of DiGeorge syndrome: prenatal detection by FISH of a molecular 22q11 deletion. AB - We report on a prenatal diagnosis by FISH of a familial 22q11 deletion associated with DiGeorge syndrome (DGS). The deletion was seen in the proband with symptoms of full DGS, in the physically normal father, and in a subsequent pregnancy. After birth this child showed hypocalcaemia, a T cell deficit, and a right sided aortic arch. PMID- 7473664 TI - Filippi syndrome: a new case with skeletal abnormalities. AB - We report on a 9 year old girl, born to consanguineous parents, with major microcephaly, cutaneous syndactyly of the toes, and moderate mental retardation with marked speech involvement. In addition, moderate dysmorphic features and skeletal abnormalities were noted. This multiple congenital anomalies/mental retardation pattern very much resembles that described by Filippi. This observation confirms that this syndrome is a distinct, probably autosomal recessive entity. PMID- 7473665 TI - Report on MDA workshop on myotonic dystrophy, 10 October 1994, Montreal, Quebec, Canada. PMID- 7473666 TI - Anophthalmia with cleft palate and micrognathia: a new syndrome or an unusual presentation of Rubinstein-Taybi syndrome? PMID- 7473667 TI - Pallister-Hall and McKusick-Kaufmann syndromes. PMID- 7473668 TI - Idiopathic hypoparathyroidism in two patients with 22q11 microdeletion. PMID- 7473669 TI - Prototype sequence clues within the Fanconi anaemia group C gene. PMID- 7473670 TI - The COX8 gene is not the disease gene of the CMH4 locus in familial hypertrophic cardiomyopathy. PMID- 7473671 TI - Tarnished gold: sixty years of antimicrobial drug use and misuse. PMID- 7473672 TI - Human immunity to rotavirus. AB - Rotaviruses are the most important cause of severe gastro-enteritis in infants and young children. However, the determinants of protective immunity are poorly understood. Human immunity to rotavirus can be acquired passively or actively. It may be humoral or cell-mediated, protective or non-protective, homotypic or heterotypic and mucosal or systemic, or any combination of these. Mucosal immunity is protective against rotavirus illness, but not against infection, whereas systemic immunity reflects exposure, but probably has little if any role in protection. Both local and cell-mediated immunity are likely to be important in protection. However, there is no agreement as to a reliable surrogate marker of small intestinal protective immunity, and little is known about small intestinal cell-mediated immunity in man, especially infants. Passive mucosal immunity, but not systemic immunity, may contribute to protection in breast-fed infants, and in those at increased risk of serious illness who have been given oral immunoglobulin, either as prophylaxis or therapeutically. Animal and adult studies may have only limited relevance to those who are at greatest risk of serious illness. However, it is probably from such studies that hypotheses about small intestinal cell-mediated immunity in the protection of infants against rotavirus infection in man remain unclear, and this continues to hinder vaccine research. PMID- 7473673 TI - Potential pathogenic properties of members of the "Streptococcus milleri" group in relation to the production of endocarditis and abscesses. AB - The "Streptococcus milleri" (SMG) group have been shown to possess factors in vitro that may be involved in pathogenesis. All SMG strains are able to bind fibronectin via a cell-surface protein; the binding ranged from 12 to 198 mol/cell. Strains also bound to platelet-fibrin or fibrin clots and fibrinogen, giving maximum adhesion values of 16.5%, 21.8% and 151 mol/cell respectively. Members of the species S. constellatus produced thrombin-like activity. Lancefield group C SMG aggregated rat platelets, a bacterial cell-surface protein acting as mediator in the reaction. Most of the in-vitro factors did not correlate with each other, an indication that SMG strains possess a wide variety of pathogenic properties that may be involved in the production of abscesses or endocarditis. However, there was a correlation between the binding of large amounts of fibrinogen ( > 100 mol/cell) and the ability to aggregate platelets. This suggests that fibrinogen binding may aid in platelet aggregation. PMID- 7473674 TI - Ultrastructural variation of cultured Ehrlichia chaffeensis. AB - The ultrastructure of Ehrlichia chaffeensis (Arkansas strain) was studied in non irradiated and irradiated monolayers of mouse embryo, Vero, BGM and L929 cells, and in non-irradiated DH82 cells. Within the intracellular parasitophorous vacuoles (morulae), two types of ehrlichial cells were found regularly--those with uniformly dispersed nucleoid filaments and ribosomes (dense-cored cells), which represent the normal life cycle of ehrlichiae. In addition, large reticulate cells were observed, forming long projections of the cell wall, protrusions of cytoplasmic membrane into the periplasmic space, or budding of protoplast fragments (minute forms) into the periplasmic space. Ehrlichiae with abnormalities of protoplast fission were found, apparently leading to formation of giant, multilobular or elongated rod-like ehrlichiae. Morulae were usually surrounded by cisterns of granular endoplasmic reticulum and mitochondria and often contained vesicles, long tubules 25nm in diameter, probably originating from the ehrlichial cell wall, and fibrillar ehrlichial antigen apparently shed from the surface of the cell wall. Some cells contained, in addition to normal morulae, a whole morula that had become dense and contained degenerating ehrlichiae. These results indicate that as well as normal growth and reproduction, ehrlichiae exhibit pathological events: they can be remarkably damaged inside the host cell vacuoles, presumably phagolysosomes, or enter a process morphologically similar to bacterial L-transformation. PMID- 7473675 TI - Molecular mimicry by Mycoplasma pneumoniae to evade the induction of adherence inhibiting antibodies. AB - Specific regions of adherence binding sites and epitopes of the P1 adhesin of Mycoplasma pneumoniae were synthesised as octapeptides and used as targets in a modified enzyme-linked immunosorbent assay. Acute phase and convalescent sera from 10 patients with M. pneumoniae infection were tested for antibody reactivity to these octapeptides. In convalescent sera, antibody activities were directed against octapeptides of the epitope regions, whereas no antibody activity was found in acute or convalescent sera to octapeptides of adherence-mediating binding sites could be explained partially from the results of cross-reactivity experiments with adherence-inhibiting anti-P1 adhesin monoclonal antibodies (MAbs). Two of these MAbs showed cross-reactions with intracellular antigens of eukaryotic cell lines in immunofluorescence microscopy experiments. The cross reacting antigens were isolated and characterised as glyceraldehyde-3-phosphate dehydrogenase and 2-phospho-D-glycerate hydrolyase. Antigenic mimicry of eukaryotic structures by functional sites of the P1 adhesin of M. pneumoniae may influence the pathogenesis of M. pneumoniae infection. PMID- 7473676 TI - Comparison of detection of human papillomavirus 16 DNA in cervical carcinoma tissues by Southern blot hybridisation and nested polymerase chain reaction. AB - An association between human papillomavirus (HPV) and cervical neoplasia has been widely reported and HPV DNA is commonly detected in cervical carcinoma tissues. However, estimates of the prevalence of HPV infection differs among various detection methods. Seventy cases of cervical carcinoma were screened for HPV 16 infection by Southern blot hybridisation (SBH) and nested polymerase chain reaction (PCR). According to SBH, the prevalences of HPV 16 DNA in stage I (n = 40) and stage II (n = 30) cervical carcinomas were 52.5 and 63.3%, respectively, and the overall prevalence was 57.1% (40 of 70). By nested PCR, the prevalences of HPV 16 infection in stage I and II cervical carcinomas were 87.5 and 93.3%, respectively, and the overall prevalence was 90.3%. The prevalence of HPV DNA detected by nested PCR was significantly greater than that detected by SBH. The combined concordance of positive and negative results between SBH and nested PCR was 61.4%. The discrepancy resulted mainly from 25 cases (35.7%) that were positive by PCR but negative by SBH. A small copy number of HPV DNA in the these 25 cases was documented by a semi-quantitative PCR method. The nested PCR was more sensitive than SBH and detected cases with low amounts of HPV DNA. The detection of HPV infection varied between these two prevailing detection methods and this should be kept in mind in assessing various epidemiological data concerning HPV infection. PMID- 7473678 TI - Enterotoxigenic Clostridium perfringens as a cause of sporadic cases of diarrhoea. AB - The purpose of this study was to investigate the incidence of cases of sporadic diarrhoea associated with enterotoxigenic Clostridium perfringens. Cases were identified by detection of C. perfringens enterotoxin with the Oxoid RPLA kit, with confirmation by ELISA, in faecal specimens from isolated incidents of diarrhoea and from which no other enteropathogen had been isolated. In a 2-month study, 65 (18%) of 370 specimens were enterotoxin positive. There was no predominant age group or sex in the enterotoxin-positive group, but higher proportion (79%) was resident in the community than were enterotoxin-negative cases (34%). Only four of the 65 enterotoxin-positive patients had received antibiotic therapy. Spore counts in most enterotoxin-positive patients were < 10(5)/g, indicating that detection of high numbers of C. perfringens is not useful in determining the aetiology of sporadic diarrhoea. Diagnosis should be confirmed by the detection of enterotoxin, but further work is required to assess whether an acceptable accuracy is obtained with the RPLA kit or whether ELISA is needed in all cases. PMID- 7473677 TI - Characterisation of Burkholderia cepacia from cystic fibrosis patients living in Wales by PCR ribotyping. AB - Polymerase chain reaction (PCR) ribotyping detects differences in the intergenic spacer between the 16S and 23SrRNA genes. This method was applied to Burkholderia cepacia isolates from 16 Welsh cystic fibrosis (CF) patients attending three different clinics. Amplification of the intergenic spacer followed by an additional digestion step with TaqI restriction endonuclease identified seven distinct electrophoretic patterns among the patient isolates. Each of the seven patterns was distinct from that of the so called "epidemic strain" commonly isolated from patients attending clinics elsewhere in the UK. Two environmental isolates from the hospital clinics and four NCTC reference strains gave different patterns. The simplicity of the method lends itself to use in a general microbiological laboratory. PMID- 7473679 TI - Automated laser fluorescence analysis of randomly amplified polymorphic DNA: a rapid method for investigating nosocomial transmission of Acinetobacter baumannii. AB - A rapid method for genotyping Acinetobacter baumannii based on PCR-fingerprinting with fluorescent primers was evaluated. Automated laser fluorescence analysis (ALFA) enabled on-line generation of high resolution DNA-fingerprints during polyacrylamide gel electrophoresis of randomly amplified polymorphic DNA (RAPD) products. The results were in concordance with macro-restriction fragment patterns produced by pulse-field gel electrophoresis (PFGE) of ApaI digests of chromosomal DNA. RAPD-ALFA was able to identify homologous strains suggestive of horizontal transmission in < 8 h after colonies were obtained on solid media, whereas PFGE analysis took c. 90 h. Speed and digitised data format renders RAPD ALFA attractive for routine in-house epidemiological screening of isolates from intensive care and other hospital units. PMID- 7473680 TI - Candida albicans isolates from HIV-infected and AIDS patients exhibit enhanced adherence to epithelial cells. AB - The increased prevalence of oral candidosis associated with HIV infection must be intrinsically related to immunological changes in the host, but might also involve alterations to the infecting strains of yeast. This study aimed to determine if strains of Candida albicans isolated from asymptomatic HIV-infected individuals or AIDS patients possessed altered adherence properties in an in vitro buccal epithelial cell (BEC) adherence assay. C. albicans isolates from 49 patients with HIV infection or AIDS adhered to BEC in significantly higher numbers than isolates from 49 control subjects (p < 0.001). No significant differences in adherence were detected between strains isolated from HIV-infected or AIDS subjects, or between strains isolated from C. Albicans carriers (low salivary C. albicans counts) or subjects with oral candidosis. The presence of whole saliva significantly inhibited the binding of candida to BEC (p < 0.001), but the significant difference in adherence between the HIV/AIDS and control isolates was maintained. The effect of saliva was independent of salivary candida antibodies and was abolished by treatment with protease or neuraminidase, suggesting the involvement of salivary mucins. The results of this study suggest that HIV infection is associated with the selection of strains of C. albicans with and increased ability to adhere to oral mucosa. PMID- 7473681 TI - Potentiation of the effects of chlorhexidine diacetate and cetylpyridinium chloride on mycobacteria by ethambutol. AB - Ethambutol enhanced the effects of chlorhexidine diacetate and cetylpyridinium chloride against Mycobacterium avium, M. bovis BCG, M. fortuitum and M. phlei. The findings show that it is possible to increase the susceptibility of mycobacteria to agents that normally exhibit poor activity against these organisms because of their reduced cellular penetration. PMID- 7473683 TI - Reconstitution of hepatic uricase in planar lipid bilayer reveals a functional organic anion channel. AB - Rat renal proximal tubule cell membranes have been reported to contain uricase like proteins that function as electrogenic urate transporters. Although uricase, per se, has only been detected within peroxisomes in rat liver (where it functions as an oxidative enzyme) this protein has been shown to function as a urate transport protein when inserted into liposomes. Since both the uricase-like renal protein and hepatic uricase can transport urate, reconstitution studies were performed to further characterize the mechanism by which uricase may function as a transport protein. Ion channel activity was evaluated in planar lipid bilayers before and after fusion of uricase-containing proteoliposomes. In the presence of symmetrical solutions of urate and KCl, but absence of uricase, no current was generated when the voltage was ramped between +/- 100 mV. Following fusion of uricase with the bilayer, single channel activity was evident: the reconstituted channel rectified with a mean slope conductance of 8 pS, displayed voltage sensitivity, and demonstrated a marked selectivity for urate relative to K+ and Cl-. The channel was more selective to oxonate, an inhibitor of both enzymatic uricase activity and urate transport, than urate and it was equally selective to urate and pyrazinoate, an inhibitor of urate transport. With time, pyrazinoate blocked both its own movement and the movement of urate through the channel. Channel activity was also blocked by the IgG fraction of a polyclonal antibody to affinity purified pig liver uricase. These studies demonstrate that a highly selective, voltage dependent organic anion channel is formed when a purified preparation of uricase is reconstituted in lipid bilayers. PMID- 7473684 TI - The gating of the sheep skeletal sarcoplasmic reticulum Ca(2+)-release channel is regulated by luminal Ca2+. AB - The effects of changes in luminal [Ca2+] have been investigated in sheep skeletal sarcoplasmic reticulum (SR) Ca(2+)-release channels after activation of the channels by different ligands from the cytosolic side of the channel. Native heavy SR membrane vesicles were incorporated into planar phospholipid bilayers under voltage-clamp conditions. Experiments were carried out in symmetrical 250 mM Cs+. Lifetime analysis indicates that channels activated solely by cytosolic Ca2+ exhibit at least two open and five closed states. The open events are very brief and are close to the minimum resolvable duration. When channels are activated solely by cytosolic Ca2+, luminal Ca2+ does not appear to exert any regulatory effect. The Po and duration of the open and closed lifetimes are unchanged. However, if channels are activated by ATP alone or by ATP plus cytosolic Ca2+, increases in luminal [Ca2+] produce marked increases in Po and in the duration of the open lifetimes. Our results demonstrate that maximum activation of the skeletal SR Ca(2+)-release channel by ATP cannot be obtained in the absence of millimolar luminal [Ca2+]. PMID- 7473682 TI - Molecular mechanisms in exocytosis. PMID- 7473686 TI - Magnitude and modulation of pancreatic beta-cell gap junction electrical conductance in situ. AB - The parallel gap junction electrical conductance between a beta-cell and its nearest neighbors was measured by using an intracellular microelectrode to clamp the voltage of a beta-cell within a bursting islet of Langerhans. The holding current records consisted of bursts of inward current due to the synchronized oscillations in membrane potential of the surrounding cells. The membrane potential record of the impaled cell, obtained in current clamp mode, was used to estimate the behavior of the surrounding cells during voltage clamp, and the coupling conductance was calculated by dividing the magnitude of the current bursts by that of the voltage bursts. The histogram of coupling conductance magnitude from 26 cells was bimodal with peaks at 2.5 and 3.5 nS, indicating heterogeneity in extent of electrical communication within the islet of Langerhans. Gap junction conductance reversibly decreased when the temperature was lowered from 37 to 30 degrees C and when the extracellular calcium concentration was raised from 2.56 to 7.56 mM. The coupling conductance decreased slightly during the active phase of the burst. Activation of adenylate cyclase with forskolin (10 microM) resulted in an increase in cell-to-cell electrical coupling. We conclude that beta-cell gap junction conductance can be measured in situ under near physiological conditions. Furthermore, the magnitude and physiological regulation of beta-cell gap junction conductance suggest that intercellular electrical communication plays an important role in the function of the endocrine pancreas. PMID- 7473687 TI - The permeation of organic cations through cAMP-gated channels in mammalian olfactory receptor neurons. AB - The permeation of monovalent organic cations through adenosine 3',5'-cyclic monophosphate-(cAMP) activated channels was studied by recording macroscopic currents in excised inside-out membrane patches from the dendritic knobs of isolated mammalian olfactory receptor neurons (ORNs). Current-voltage relations were measured when bathing solution Na+ was replaced by monovalent organic cations. Permeability ratios relative to Na+ ions were calculated from changes in reversal potentials. Some of the small organic cations tested included ammonium (NH4+), hydroxylammonium and formamidinium, with relative permeability ratios of 1.41, 2.3 and 1.01 respectively. The larger methylated and ethylated ammonium ions studied included: DMA (dimethylammonium), TMA (tetramethylammonium) and TEA (tetraethylammonium) and they all had permeability ratios larger than 0.09. Even large cations such as choline, arginine and tris(hydroxymethyl)aminomethane (Tris) were appreciably permeant through the cAMP-activated channel with permeability ratios ranging from 0.19 to 0.7. The size of the permeating cations, as assessed by molecular weight, was a good predictor of the permeability. The permeability sequence of the cAMP-activated channel in our study was PNH4 > PNa > PDMA > PTMA > PCholine > PTEA. Higher permeability ratios of hydroxylammonium, arginine and tris(hydroxymethyl)aminomethane cannot be explained by ionic size alone. Our results indicate that: (i) cAMP-activated channels poorly select between monovalent cations; (ii) the pore dimension must be at least 6.5 x 6.5 A, in order to allow TEA and Tris to permeate and (iii) molecular sieving must be an important mechanism for the permeation of large organic ions through the channels with specific ion binding playing a smaller role than in other structurally similar channels. In addition, the results clearly indicate that cyclic nucleotide-gated (CNG) channels in different cells are not the same, the olfactory CNG channel being different from that of the photoreceptors, particularly with respect to the permeation of large organic cations, which the ORN channels allow to permeate readily. PMID- 7473685 TI - Effects of cysteine modification on the activity of the cGMP-gated channel from retinal rods. AB - The effect of sulfhydryl reagents on the activity of the cGMP-gated channel from bovine retinal rods was studied by measurements of 8-Br-cGMP-(cGMP)-induced calcium efflux from rod membrane vesicles and records of 8-Br-cGMP-dependent sodium currents through channels incorporated into planar lipid bilayers. N ethylmaleimide and mersalyl (thiol blockers) as well as diamide (dithiol disulfide conversion agent) have a dual effect on the channels activity: at low concentration, they increase the apparent affinity for cyclic nucleotide ("activation") at the same time inducing a loss of cooperativity for nucleotide binding; at higher concentration, N-ethylmaleimide and diamide produce a reduction of the amplitude and initial rate of the calcium release at saturating nucleotide concentration, while mersalyl is shown to reduce the activity of the channels in bilayer experiments ("inhibition"). Nitric oxide precursors have no effect. The results suggest that blocking at least 1 of the 3 cytoplasmic cysteine residues situated close to the cGMP-binding site in each channel subunit by N-ethylmaleimide, mersalyl, or diamide (forming a dimer between 2 subunits) increases the affinity for the nucleotide. Inhibition is produced by blocking at least one of the 2 other cytoplasmic sulfhydryl groups (N-ethylmaleimide, mersalyl, oxidized glutathione) or the 2 others (diamide, intrasubunit bridge), and may concern a process of channel inactivation. The 3 cytoplasmic sulfhydryl groups are accessible when the channels are in the open state, but not (or much less) accessible when the channels are in the closed state. PMID- 7473688 TI - Relationship of alcohol-induced changes in Mg(2+)-ATPase activity of rabbit intestinal brush border membrane with changes in fluidity of its lipid bilayer. AB - We examined the effects of seven n-alkyl alcohols (from n-butyl to n-undecyl alcohol), isoamyl alcohol and benzyl alcohol on the activity of membrane enzyme Mg(2+)-ATPase of the rabbit small intestinal brush border membrane. Their relationships with the changes in the fluidity of the membrane lipid bilayer were examined through studies on the fluorescence anisotropies of diphenylhexatriene (DPH) and its ionic derivatives. Good linear correlations were found both between the partition coefficients of the alcohols and their concentrations causing similar decreases in the activity of Mg(2+)-ATPase and between their partition coefficients and the alcohol-induced changes in fluorescence anisotropies. Within the concentration range of the alcohols tested, including isoamyl alcohol and benzyl alcohol, the decreases in activity of the membrane enzyme Mg(2+)-ATPase clearly corresponded with the decreases in fluorescence anisotropy of DPH, which is thought to be located within the hydrophobic core of the membrane. From these findings, one possible explanation is that inhibition of this enzyme by the alcohols is due to perturbation of the lipid bilayer of the brush border membrane. PMID- 7473689 TI - Calcium-sensitivity of the plasmalemmal delayed rectifier potassium current suggests that calcium influx in pulvinar protoplasts from Mimosa pudica L. can be revealed by hyperpolarization. AB - Isolated protoplasts from pulvinar motor cells of Mimosa pudica were studied using conventional whole-cell patch clamp techniques. With internal solutions weakly buffered for Ca2+ (0.2 mM EGTA), a rundown of the outward delayed rectifier K+ current was induced by hyperpolarizing the holding potential, and this effect was strongly promoted by high external Ca2+ concentrations. This rundown could be reversed by coming back to less hyperpolarized holding potentials or by lowering the external [Ca2+]. Such rundown was absent when pipette internal solutions strongly buffered (10 mM EGTA) for Ca2+ were used. Ionomycin induced rundown of the K+ current with internal solutions containing 0.2 mM but not 10 mM EGTA. The hyperpolarization-associated rundown was reversibly blocked by Gd3+ and La3+. PMID- 7473692 TI - A non-invasive photographic modification for recording early vascular development in the chick. AB - The chick embryo shell-less culture technique allows continuous observation and access to the developing vessels of the area vasculosa (AV); hence, its value as an angiogenesis assay system. The main drawback of the method is that adequately contrasted photographs cannot be achieved without injection of a contrast medium which kills the embryo making further observation of the same specimen impossible. Furthermore, injection prior to 72 h incubation has a high failure rate. In an attempt to overcome these problems, we explored the possibility of using a high-contrast photographic process. Embryos were explanted into shell less culture after 48 h incubation and photographed through a stereo microscope. After trials with different films we found that Kodalith ortho Type 3 (Kodak) and Technical Pan film (Kodak) produced images of the vasculature which were identical in contrast and detail to India-ink-injected specimens photographed with conventional film. In addition, excellent images could be recorded as early as 48 h of incubation. PMID- 7473691 TI - Double-layer coating for high-resolution low-temperature scanning electron microscopy. AB - Specimen damage caused by mass loss due to electron beam irradiation is a major limitation in low-temperature scanning electron microscopy of bulk specimens. At high primary magnifications (e.g., 100,000 x) a hydrated sample is usually severely damaged after one slow scan (about 3000 e-nm-2). The consequences of this beam damage are significantly reduced by coating the frozen-hydrated sample with a 5-10-nm-thick carbon layer. Since this layer covers up surface details, the sample is first unidirectionally shadowed with a thin heavy metal layer (e.g., 2 nm of platinum) that is in close contact with the biological surface (double layer coating). This heavy metal layer can be visualized in field emission scanning electron microscopy with the material-dependent backscattered electron signal. The method allows for routine observation of large frozen hydrated samples. By use of an in-lens field-emission SEM and a sensitive backscattered electron detector, structural information comparable to that obtained with the transmission electron microscopy freeze-fracture replica technique can be achieved. PMID- 7473690 TI - Lumenal calcium modulates unitary conductance and gating of a plant vacuolar calcium release channel. AB - The patch clamp technique has been used to investigate ion permeation and Ca(2+) dependent gating of a voltage-sensitive Ca2+ release channel in the vacuolar membrane of sugar beet tap roots. Reversal potential measurements in bi-ionic conditions revealed a sequence for permeability ratios of Ca2+ approximately Sr2+ approximately Ba2+ > Mg2+ >> K+ which is inversely related to the size of the unitary conductances K+ >> Mg2+ approximately Ba2+ > Sr2+ approximately Ca2+, suggesting that ion movement is not independent. In the presence of Ca2+, the unitary K+ current is reduced in a concentration- and voltage-dependent manner by Ca2+ binding at a high affinity site (K0.5 = 0.29 mM at 0 mV) which is located 9% along the electric field of the membrane from the vacuolar side. Comparison of reversal potentials obtained under strictly bi-ionic conditions with those obtained in the presence of mixtures of the two ions indicates that the channel forms a multi-ion pore. Lumenal Ca2+ also has an effect on voltage-dependent channel gating. Stepwise increases of vacuolar Ca2+ from micromolar to millimolar concentrations resulted in a dramatic increase in channel openings over the physiological voltage range via a shift in threshold for channel activation to less negative membrane potentials. The steepness of the concentration dependence of channel activation by Ca2+ at -41 mV predicts that two Ca2+ ions need to bind to open the gate. The implications of the results for ion permeation and channel gating are discussed. PMID- 7473693 TI - Signal standardization of the secondary ion mass spectrometry (SIMS) microscope for quantification of halogens and calcium in biological applications. AB - The secondary ion mass spectrometry (SIMS) microscope is able to map chemical elements in tissue sections. Although absolute quantification of an element remains difficult, a relative quantitative approach is possible for soft tissue by using carbon (12C) as an internal reference present at large homogeneous and constant concentration in specimen and embedding resin. In this study, this approach is used to standardize the signal of an SIMS microscope for the quantification of halogens (9F-, 35Cl- and 79Br-) and calcium (40Ca+). Standard preparation was determined based on homogeneity and stability criteria by molecular incorporation (halogens) or mixing (calcium) in methacrylate resin. Standard measurements were performed by depth analysis on areas of 8 microns (halogens) and 150 microns (calcium) in diameter for 10-30 min, under Cs+ (halogens) or Ox+ (calcium) bombardment. Results obtained from 100-120 measurements for each standard dilution show that the relationship between the signal intensity measured and the elemental concentration (micrograms/mg of wet tissue or mM) is linear in the range of biological concentrations. This quantitative approach was applied firstly to bromine of the 5-bromo-2' deoxyuridine (BrdU) used as nuclear marker of rat hepatocytes in proliferation. The second model concerns depletion of calcium concentration in cortical compartment in Paramecium tetraurelia during exocytosis. Then signal standardization in SIMS microscopy allows us to correlate quantitative results with those obtained from other methods. PMID- 7473694 TI - Vitrification of articular cartilage by high-pressure freezing. AB - For more than 20 years, high-pressure freezing has been used to cryofix bulk biological specimens and reports are available in which the potential and limits of this method have been evaluated mostly based on morphological criteria. By evaluating the presence or absence of segregation patterns, it was postulated that biological samples of up to 600 microns in thickness could be vitrified by high-pressure freezing. The cooling rates necessary to achieve this result under high-pressure conditions were estimated to be of the order of several hundred degrees kelvin per second. Recent results suggest that the thickness of biological samples which can be vitrified may be much less than previously believed. It was the aim of this study to explore the potential and limits of high-pressure freezing using theoretical and experimental methods. A new high pressure freezing apparatus (Leica EM HPF), which can generate higher cooling rates at the sample surface than previously possible, was used. Using bovine articular cartilage as a model tissue system, we were able to vitrify 150-micron thick tissue samples. Vitrification was proven by subjecting frozen-hydrated cryosections to electron diffraction analysis and was found to be dependent on the proteoglycan concentration and water content of the cartilage. Only the lower radical zone (with a high proteoglycan concentration and a low water content compared to the other zones) could be fully vitrified. Our theoretical calculations indicated that applied surface cooling rates in excess of 5000 K/s can be propagated into specimen centres only if samples are relatively thin (< 200 microns). These calculations, taken together with our zone-dependent attainment of vitrification in 150-micron-thick cartilage samples, suggest that the critical cooling rates necessary to achieve vitrification of biological samples under high-pressure freezing conditions are significantly their (1000 100,000 K/s) than previously proposed, but are reduced by about a factor of 100 when compared to cooling rates necessary to vitrify biological samples at ambient pressure. PMID- 7473695 TI - Cervical discogenic syndrome: a cause of chronic head and neck pain. AB - In patients with persistent pain of the neck, head, shoulder, and periscapular area, the diagnosis of cervical discogenic syndrome should be considered. The intervertebral disc can be a source of persistent pain and the severity of the symptoms should dictate the treatment. Surgical intervention should only be considered as a last resort in someone with significant pain who has failed prolonged conservative therapy. The success of the surgical treatment of this entity is most closely linked to adequate patient selection and rigorous patient preparation. PMID- 7473696 TI - DNA ploidy of thyroid lesions. PMID- 7473697 TI - Transcript: remarks on Medicare reform from the AMA House of Delegates' annual meeting. PMID- 7473698 TI - Quality time. PMID- 7473699 TI - Cardiovascular disease in Mississippi. PMID- 7473700 TI - Physical therapy as an effective change agent in the treatment of patients with urinary incontinence. PMID- 7473701 TI - You've come a long way, baby! PMID- 7473702 TI - AMA applauds President's call for curbs on teen use of tobacco. PMID- 7473703 TI - Mid-life crisis at age thirty. PMID- 7473704 TI - Delegate's AMA report 1995 annual meeting of the House of Delegates. PMID- 7473705 TI - Effects of base substitutions on the binding of a DNA-bending protein. AB - In order to investigate whether the 2-amino group of guanine, which lies in the minor groove of the B-form helix, can directly influence DNA flexibility and major groove recognition by proteins we have examined the properties of DNA molecules containing inosine and/or 2,6-diaminopurine (DAP) residues. Appropriately substituted tyrT(A93) DNA fragments were prepared by the polymerase chain reaction. Their mobility in non-denaturing gels was affected, consistent with changed anisotropic flexibility leading to increased curvature due to G-->I substitution and decreased curvature due to replacement of adenine with DAP. Band shift assays of FIS protein binding revealed facilitated interaction with inosine containing DNA and markedly reduced binding to DAP-containing DNA, attributable to altered bendability. DNase footprinting experiments confirmed that fewer sites would bind FIS in DAP-containing DNA at a given protein concentration, whereas higher levels of binding occurred with inosine-containing molecules. Thus base substitutions which affect the placement and presence of the purine 2-amino group in the minor groove can affect both the intrinsic curvature and the bendability of DNA. PMID- 7473706 TI - Solution structure of the anticodon-binding domain of Escherichia coli lysyl-tRNA synthetase and studies of its interaction with tRNA(Lys). AB - A protein domain corresponding to residues 31 to 149 of the E. coli Lysyl-tRNA synthetase species corresponding to the lysS gene was expressed and 15N-labelled. 1H and 15N NMR resonance assignments for this domain were obtained by two dimensional and three-dimensional homonuclear and heteronuclear spectroscopy. Using distance geometry and simulated annealing, a three-dimensional structure could be calculated using 701 NOE and 86 dihedral angle restraints. It is composed of a five-stranded antiparallel beta-barrel capped by three alpha helices at its ends. This structure closely resembles that of the N-terminal domain of the other E. coli lysyl-tRNA synthetase species expressed from the lysU gene and is highly homologous to the fold observed for the corresponding region of aspartyl-tRNA synthetase. It is shown that the isolated N-terminal fragment of lysyl-tRNA synthetase can interact with tRNA(Lys) as well as with poly (U), which mimics the anticodon sequence. Amino acid residues involved in these interactions were identified and, in the case of poly-U, a number of specific protein-RNA contacts were characterized. Specific recognition of tRNA(Lys) involves a cluster of four structurally well-defined aromatic residues, anchored on the beta strands, and basic residues located on the surrounding loops. This organization is reminiscent of other RNA binding proteins, such as the U1A small nuclear ribonucleoprotein. PMID- 7473707 TI - Prediction of protein three-dimensional structures in insertion and deletion regions: a procedure for searching data bases of representative protein fragments using geometric scoring criteria. AB - The prediction of protein structure in insertion/deletion regions (referred to as indels) is an important part of protein model building by homology. Here we combine cluster analysis with data base search procedures. Initially, data bases of representative protein fragments are constructed using two different clustering algorithms. In the HCAPD (hierarchical clustering after preliminary division) approach, all protein fragments are divided into classes with similar anchor region structures (a protein fragment consists of two anchoring regions and a central region). Within these classes the fragments are further clustered using a hierarchical cluster algorithm. The DCANN (deterministic clustering by assignment of all nearest neighbours) approach is a variant of the k-nearest neighbours cluster algorithm. Only geometric scoring criteria are used for data base searching. The main advantage of a non-redundant data base is the ability to provide structurally different fragments during the search process, which leads to an improvement in structure prediction. Both methods have been tested on 71 insertions and 74 deletions with lengths between one and eight residues. PMID- 7473708 TI - X-ray crystal structure analysis of the catalytic domain of the oncogene product p21H-ras complexed with caged GTP and mant dGppNHp. AB - The X-ray structures of the 1:1 complexes formed between p21H-ras (residues 1 to 166) and the nucleotides P3-1-(2-nitrophenyl)ethyl guanosine triphosphate ("caged GTP"; pure R- and S-diastereomers) and 3'-O-(N-methylanthraniloyl)-2' deoxyguanosine 5'-(beta, gamma-imido)-triphosphate ("mant dG-ppNHp"), have been refined to an R-factor of 21.4% (R-caged GTP, 1.85 A resolution), 18.9% (S-caged GTP, 2.5 A resolution) and 17.6% (mant dGppNHp, 2.7 A resolution), respectively. Details of the structure determination, refinement and the structures themselves are presented. The overall structures of the complexes are identical in terms of the general organization of their secondary structure elements and are also identical to that reported for the analogous complex of p21H-ras with GppNHp. The binding of the GTP part is not significantly affected by the additional aromatic group (cage and mant, respectively) in contrast to the original observation on p21:caged GTP using the racemic mixture of R- and S-caged GTP. The main differences in the structures are observed in the region of loop L2 (residues Glu31 to Thr35) where the additional aromatic group attached to the nucleotide comes very close to the side-chain of Tyr32, including backbone displacements of 2.6 A, 2.2 A and 0.3 A for the residues from Glu31 to Thr35 for R-caged, S-caged GTP and mant dGppNHp, respectively. The refined structures provide additional data for the design of new nucleotide analogs and the importance of their stereochemistry as well as for the design of new mutant forms of p21H-ras for further biochemical investigations. The binding mode of mant dGppNHp reveals significant features for the understanding of the fluorescence signals observed in solution. PMID- 7473709 TI - Studies on the lumazine synthase/riboflavin synthase complex of Bacillus subtilis: crystal structure analysis of reconstituted, icosahedral beta-subunit capsids with bound substrate analogue inhibitor at 2.4 A resolution. AB - The lumazine synthase/riboflavin synthase of Bacillus subtilis is a bifunctional enzyme complex catalysing the formation of riboflavin from 5-amino-6-(D ribitylamino)-2,4(1H,3H)-pyrimidinedione and L-3,4-dihydroxy-2-butanone-4 phosphate via 6,7-dimethyl-8-ribityllumazine. The complex is composed of 3 alpha (riboflavin synthase) subunits and 60 beta (lumazine synthase) subunits and has a relative mass of 1 MDa. The 60 beta subunits are arranged in an icosahedral capsid enclosing the alpha trimer in the central core. The protein was crystallised, and an X-ray structure of the icosahedral capsid was obtained at 3.3 A resolution with a crystallographic R-factor of 0.33. Hollow, icosahedral capsids consisting of 60 beta subunits can be obtained by inhibitor-driven renaturation of isolated beta subunits. They catalyse the formation of 6,7 dimethyl-8-ribityllumazine at the same rate as the native alpha 3 beta 60 complex and can be crystallised in two different hexagonal and one monoclinic form. Crystallographic intensity data of the monoclinic crystals to a resolution of 2.4 A were obtained using synchrotron radiation and an image plate detector system. The orientation of the icosahedral molecules in the monoclinic cell was deduced by real space vector search procedures from a 3.5 A Patterson map. Phases were calculated from the model of the alpha 3 beta 60 protein and were extended by cyclic averaging exploring the 30-fold redundancy of the electron density. The 2.4 A map allowed us to refine the existing atomic model of lumazine synthase. The refined model includes 154 amino acid residues, one inhibitor molecule, 58 water molecules and one phosphate ion. Applying non-crystallographic-symmetry restraints the crystallographic R-factor is 16.7% for 100,092 reflections between 10 and 2.4 A. The chain folding of the beta subunits is closely similar to the native alpha 3 beta 60 enzyme. The lumazine synthase bears resemblance to the sugar binding proteins. The significantly higher resolution compared to the alpha 3 beta 60 structure determination allows a detailed description of the substrate analogue binding site. The environment of the 5-nitro-6-(D-ribitylamino) 2,4(1H,3H)-pyrimidinedione inhibitor is particularly rigid, and the chain segments involved in forming the active site are highly conserved for lumazine synthases of different species. A residual density feature in the final map is interpreted as a bound phosphate which mimics the binding of the second substrate. We discuss the reaction mechanism on this structural basis. PMID- 7473710 TI - The 2.0 A crystal structure of Scapharca tetrameric hemoglobin: cooperative dimers within an allosteric tetramer. AB - The crystal structure of the allosteric tetrameric hemoglobin from Scapharca inaequivalvis (HbII) has been determined in the carbonmonoxy liganded state using a combination of anomalous scattering and molecular replacement. The molecular model has been refined at 2.0 A resolution to a conventional R-factor of 0.173 and a free R-factor of 0.244. The tetramer is formed from two identical heterodimers. Each heterodimer is assembled with intersubunit contacts involving the E and F helices and heme groups in a manner that is very similar to that of the cooperative Scapharca homodimeric hemoglobin. In addition, the ordered water structure observed in these dimeric interfaces is quite similar. These structural similarities strongly suggest that the dimers within the Scapharca tetramer are cooperative. Subunits assemble into a tetramer in a distinctly non-tetrahedral arrangement, with the pseudo 2-fold axes of the heterodimer oriented at an angle of 74.5 degrees relative to the molecular 2-fold. This arrangement requires that two subunit types have distinct locations and contacts, despite the very similar tertiary structures. HbII polymerizes to higher-order assemblages in a ligand, proton and anion dependent fashion. The lattice contacts in the HbII-CO crystal suggest possible modes for this association. PMID- 7473711 TI - Myotube-specific activity of the human aldolase A M-promoter requires an overlapping binding site for NF1 and MEF2 factors in addition to a binding site (M1) for unknown proteins. AB - The human aldolase A gene is expressed in several tissues through the use of three alternative promoters. The activity of one of the promoters, pM, is restricted to skeletal muscle. We reported previously that a proximal 280 bp pM fragment confers tissue-specific expression to a CAT reporter gene in transgenic mice. This small regulatory region directs expression to muscle composed mainly of fast-twitch fibers. Here we show that a minimal promoter fragment from base pairs -164 to +45 is sufficient to highly active pM during myoblast differentiation in cell culture and demonstrate that two DNA elements play a major role in this activation. These elements consist of a binding site (M1) for unknown ubiquitous proteins and an overlapping binding site for MEF2 and NF1 families of transcription factors. The NF1 factor constitute the main binding activity on the MEF2/NF1 site and, interestingly, some of the DNA-protein complexes that form with muscle nuclear extracts on the NF1 element differ from those that form with non-muscular extracts. PMID- 7473712 TI - Refined crystal structure of recombinant murine interferon-beta at 2.15 A resolution. AB - The crystal structure of recombinant murine interferon-beta (reMuIFN-beta) has been refined at 2.15 A resolution using newly collected synchrotron data. Based on 11,228 reflections (8.0 to 2.15 A), a final R-factor of 19.1% (with a free R factor of 25.8%) was obtained with a model obeying standard geometry within 0.013 A in bond lengths and 1.4 degrees in bond angles. Compared with the previously reported model, several amino acid residues in helix A are frame-shifted, the conformations are changed for parts of loops AB and BC, helix C is extended and a new short helix exists in loop CD. Evolutionary considerations taken together, the type I interferons appear to share common structural features with respect to the chain-folding topology and the hydrogen-bond networks between various polypeptide segments. Specifically, the disposition of the C-terminal segment of loop AB (after Arg33), known to be an important receptor-binding site, seems to be strictly maintained among the type I interferons. The exposed amino acid residues on helices A and C, which have recently been implicated as the binding site for another receptor molecule, are less well conserved. This may be responsible for varied cellular effects among the subtypes of type I interferons. PMID- 7473713 TI - Elucidation of crystal packing by X-ray diffraction and freeze-etching electron microscopy. Studies on GTP cyclohydrolase I of Escherichia coli. AB - A monoclinic crystal modification of GTP cyclohydrolase I (space group P2(1), a = 204.2 A, b = 210.4 A, c = 71.8 A, alpha = gamma = 90 degrees, beta = 95.8 degrees) was studied by freeze-etching electron microscopy and by Patterson correlation techniques. The freeze-etched samples were either shadowed with Pt/C or decorated with monolayers of gold, silver or platinum. Correlation averaged electron micrographs of decoration replicas indicated 5-fold molecular symmetry. In conjunction with the molecular mass of the active GTP cyclohydrolase I enzyme complex of about 210,000 Da, which had been reported in the literature, and a molecular mass of the protomers of 24,700 Da, the electron microscopic observation suggests that the enzyme is a decamer with 5-fold symmetry. The processed images of decorated crystal surfaces also showed that the four protein multimers in the crystal unit cell are related by 4-fold pseudosymmetry. A Patterson analysis of the X-ray data showed two non-crystallographic 5-fold axes, inclined at 12 degrees to each other, thus confirming and extending the electron microscopic findings. Additionally, local 2-fold axes were found in planes perpendicular to the 5-fold particle axes. Thus, the combined X-ray and electron microscope data indicate that GTP cyclohydrolase I is a decamer with D5 symmetry. A procedure for hkl assignments of the crystal planes observed in electron micrographs was developed. On this basis, it was possible to determine the approximate molecular positions in the ab plane. Independent information on the crystal packing was obtained by single isomorphous replacement and electron density averaging. The 5-fold averaged 6 A electron density shows that the GTP cyclohydrolase I decamer is torus-shaped with an approximate diameter of 100 A and a thickness of 65 A. The study demonstrates that the combination of freeze etching electron microscopy with Patterson analysis of X-ray data is a powerful approach for the solution of complex crystallographic problems. The procedure for this analysis as well as possible pitfalls are discussed in detail. PMID- 7473714 TI - High angle and ligand-induced low angle DNA bends incited by OccR lie in the same plane with OccR bound to the interior angle. AB - The OccR protein of Agrobacterium tumefaciens activates transcription of the occQ operon in response to octopine. Octopine shortens the DNase I footprint of OccR and relaxes an OccR-incited DNA bend. In this study, we used hydroxyl radical footprinting to show that OccR contacts only one face of the operator. This contact spans 50 nucleotide pairs in the absence of octopine, and 39 nucleotide pairs in the presence of octopine. Octopine enhanced protection of the central region of the footprint. OccR blocked intercalation by methidiumpropyl EDTA along most of the operator, but did not block intercalation at the bend center. OccR protected DNA against both reagents far more completely at the promoter-distal half of the operator than at the promoter-proximal half. Experiments to alter the phasing between the operator bend and an adjacent sequence-directed bend indicated that the high angle DNA bend in the absence of octopine, and the low angle bend in the presence of octopine, lie in the same plane. Operator DNA was bent toward the helical face that OccR protected from hydroxyl radicals, indicating that OccR occupies the interior angle of this bend. PMID- 7473715 TI - Purification of KorA protein from broad host range plasmid RK2: definition of a hierarchy of KorA operators. AB - The KorA protein of promiscuous plasmid RK2 is encoded in the central control operon, which coordinates expression of genes for replication, transfer and partitioning. KorA is known to repress transcription from seven promoters on the plasmid genome but at the trfA promoter, for the vegetative replication genes, it also causes derepression of trbAp, the first promoter for the operon whose genes are required primarily for mating pair formation prior to conjugative transfer. We have overproduced and purified KorA (101 amino acid residues). Crosslinking indicates that it exists largely as a dimer in solution. Western blotting with specific antibodies suggests that there are approximately 4000 monomers per cell in exponential phase and about 600 in stationary phase. Footprinting confirmed the expected location of the KorA operator, and indicated that KorA and RNA polymerase can bind simultaneously in promoter regions. Gel retardation experiments with DNA fragments carrying each of the seven KorA operators revealed that there is a hierarchy of binding affinities. Highest affinity (Kapp = 12 to 20 nM) occurs with operators containing the 12 bp inverted repeat GTTTAGCTAAAC (klaAp, korAp and trfAp), while lower affinities (Kapp = 136 to 272 nM) occur with less perfect repeats (klcA, kleA, kleC, kfrA). In addition, specific DNA sequences flanking the 12 bp are necessary for the characteristic type I KorA binding affinity. This hierarchy may be important in providing a graded response in expression of the operons controlled by KorA as its concentration varies, as for example on transition from exponential to stationary phase. PMID- 7473716 TI - Distinctive sequence features in protein coding genic non-coding, and intergenic human DNA. AB - We have studied the behavior of a number of sequence statistics, mostly indicative of protein coding function, in a large set of human clone sequences randomly selected in the course of genome mapping (randomly selected clone sequences), and compared this with the behavior in known sequences containing genes (which we term genic sequences). As expected, given the higher coding density of the genic sequences, the sequence statistics studied behave in a substantially different manner in the randomly selected clone sequences (mostly intergenic DNA) and in the genic sequences. Strong differences in behavior of a number of such statistics are also observed, however when the randomly selected clone sequences are compared with only the non-coding fraction of the genic sequences, suggesting that intergenic and genic non-coding DNA constitute two different classes of non-coding DNA. By studying the behavior of the sequence statistics in simulated DNA of different C+G content, we have observed that a number of them are strongly dependent on C+G content. Thus, most differences between intergenic and genic non-coding DNA can be explained by differences in C+G content. A+T-rich intergenic DNA appears to be at the compositional equilibrium expected under random mutation, while C+G richer non-coding genic DNA is far from this equilibrium. The results obtained in simulated DNA indicate, on the other hand, that a very large fraction of the variation in the coding statistics that underlie gene identification algorithms is due simply to C+G content, and is not directly related to protein coding function. It appears, thus, that the performance of gene-finding algorithms should be improved by carefully distinguishing the effects of protein coding function from those of mere base compositional variation on such coding statistics. PMID- 7473717 TI - Structural studies on human rhinovirus 14 drug-resistant compensation mutants. AB - Structures have been determined of three human rhinovirus 14 (HRV14) compensation mutants that have resistance to the antiviral capsid binding compounds WIN 52035 and WIN 52084. In addition, the structure of HRV14 is reported, with a site directed mutation at residue 1219 in VP1. A spontaneous mutation occurs at the same site in one of the compensation mutants. Some of the mutations are on the viral surface in the canyon and some lie within the hydrophobic binding pocket in VP1 below the ICAM footprint. Those mutant virus strains with mutations on the surface bind better to cells than does wild-type virus. The antiviral compounds bind to the mutant viruses in a manner similar to their binding to wild-type virus. The receptor and WIN compound binding sites overlap, causing competition between receptor attachment and antiviral compound binding. The compensation mutants probably function by shifting the equilibrium in favor of receptor binding. The mutations in the canyon increase the affinity of the virus for the receptor, while the mutations in the pocket probably decrease the affinity of the WIN compounds for the virus by reducing favorable hydrophobic contacts and constricting the pore through which the antiviral compounds are thought to enter the pocket. This is in contrast to the resistant exclusion mutants that block compounds from binding by increasing the bulk of residues within the hydrophobic pocket in VP1. PMID- 7473718 TI - Assembly in vitro of bacteriophage HK97 proheads. AB - Bacteriophage HK97 is a lambdoid phage with a head assembled from 415 copies of a 42 kDa subunit arranged in an icosahedrally symmetrical lattice with a triangulation number of 7. Prohead I, the first shell structure in the assembly pathway, is composed of 42 kDa coat protein subunits that have not yet undergone the proteolytic cleavage, conformational changes, and covalent cross-linking steps that occur later in the assembly of mature heads. Prohead I can be efficiently dissociated into capsomeres by treatment with 2 M KCl. The resulting capsomeres are a mixture of two species, identified as pentamers and hexamers of the 42 kDa subunit. These capsomeres were also detected as the products of chaperonin-assisted renaturation of 42 kDa polypeptide in vitro at room temperature or in the course of self folding and assembly in vitro at 0 degrees C. Pentamer and hexamer capsomeres can be interconverted in vitro by manipulating solvent conditions, and this makes it possible to carry out the in vitro shell assembly reaction at different input ratios of hexamer to pentamer. The Prohead I structures produced are always the normal (T = 7) size regardless of the input pentamer to hexamer ratio. Assembly is most efficient when the pentamer to hexamer ratio is 1:5 (a mass ratio of 1:6), or the same as the capsomere ratio in a T = 7 shell. PMID- 7473719 TI - Sequence-specific interactions between promoter DNA and the RNA polymerase sigma factor E. AB - In order to determine which amino acyl residues in a secondary sigma factor govern its specificity of recognition at the -35 region of promoters, we examined the effects of amino acid substitutions in sigma E in Bacillus subtilis that made the sequence of its putative -35 recognition region more similar to another sigma factor in B. subtilis, sigma K. We found that a single amino acid substitution at position 217 of sigma E resulted in a sigma factor that could direct transcription from sigma K-dependent promoters. Furthermore, we tested whether this amino acid substitution in sigma E had changed the specificity of interactions of the sigma with -35 region sequences by examining the activity of the mutant sigma E on derivatives of sigma E-dependent promoters that contained single base-pair substitutions. We found that this substitution in sigma E specifically suppressed the effect of a single base-pair substitution at position -31 in a sigma E-dependent promoter spoIIID. The amino acyl residue at another position (219) on sigma E affected the specificity of interaction with position 33 in spoIIID promoter. The amino acyl residues at the two positions in sigma E, 217 and 219, that determine the specificity of interactions between the sigma and base-pairs in the -35 region of its cognate promoters (positions -33 and -31, respectively, in the spoIIID promoter) probably closely contact these base-pairs. PMID- 7473720 TI - Proteolytic and conformational control of virus capsid maturation: the bacteriophage HK97 system. AB - Bacteriophage capsid assembly pathways provide excellent model systems to study large-scale conformational changes and other mechanisms that regulate the formation of macromolecular complexes. These capsids are formed from proheads: relatively fragile precursor particles which mature by undergoing extensive remodeling. Phage HK97 employs novel features in its strategy for building capsids, including assembly without a scaffolding protein, and the formation of a network of covalent cross-links between neighboring subunits in the mature virion. In addition, proteolytic cleavage of the capsid protein from 42 kDa to 31 kDa is essential for maturation. To investigate the structural bases for proteolysis and cross-linking, we have used cryo-electron micrographs to reconstruct the three-dimensional structures of purified particles from four discrete stages in the assembly pathway: Prohead I, Prohead II, Head I and Head II. Prohead I has icosahedral T = 7 packing of blister-shaped pentamers and hexamers. The pentamers are 5-fold symmetric, but the hexamers exhibit an unusual departure from 6-fold symmetry, as if two trimers had undergone a shear dislocation of about 25 A. Proteolytic conversion to Prohead II leaves the outer surface largely unchanged, but a major loss of density from the inner surface is observed, which we infer to represent the excision of the amino-terminal domains of the capsid protein. Upon expansion to the Head I state, the capsid becomes markedly larger, thinner walled, and more polyhedral: moreover, the capsomer shapes change radically; especially notable is the disappearance of the large hexon dislocation. No differences between Head I and the covalently cross-linked Head II could be observed at the current resolution of about 25 A, from which we infer that it is the conformational rearrangements effected by expansion that create the micro-environments needed for the autocatalytic formation of the isodipeptide bonds found in the mature virions ("pseudo-active sites"). PMID- 7473721 TI - Framework residues 71 and 93 of the chimeric B72.3 antibody are major determinants of the conformation of heavy-chain hypervariable loops. AB - Structural analysis derived from the crystallographic study of the chimeric B72.3 antibody illustrated that some heavy-chain framework residues having atomic interactions with heavy-chain CDR residues may directly affect the conformation of CDR loops. For example, an alanine residue at H71 provides room for packing CDR2/CDR1 and lysine residues at H73 and H93 contribute a salt-bridge to aspartic acid at H55 in CDR2 and a hydrogen bond to the carbonyl group at H96 in CDR3, respectively. We have analysed the contribution of these framework residues to the TAG72-binding affinity. We altered these framework residues by site-directed mutagenesis, and determined the affinity of these mutant chimeric antibodies for the TAG72 antigen by solid phase radioimmunoassay. We found that a single amino acid substitution of alanine by phenylalanine at H71 or lysine by isoleucine at H93, significantly reduced the binding affinity for the TAG72 antigen by 12 and 20-fold, respectively, whereas the substitution of lysine by alanine at H73 reduced the binding affinity only two-fold. Our results indicate that heavy-chain framework residues alanine at H71 and lysine at H93 of the chimeric B72.3 antibody are the major determinants of the conformation of heavy-chain CDR2/CDR1 and CDR3 loops, whereas the salt-bridge between lysine at H73 and aspartic acid at H55 is less important. The hydrogen bond between two framework residues, glutamine at H5 and serine at H25 does not affect any CDR conformation. Our results will thus be of importance especially when the humanized B72.3 antibody is constructed by grafting the CDR loops to a human framework. The important framework region interactions must be maintained in the final humanized antibody. PMID- 7473722 TI - A simple protocol for identification of helical and mobile residues in membrane proteins. AB - A simple molecular dynamics (MD) simulations is protocol shown to predict whether a residue is in a structured alpha-helical segment or in a mobile loop or terminal segment of a membrane protein. The results are verified by comparisons with experimental NMR data. The protocol consists of performing several independent MD simulations on a polypeptide sequence of interest in a dielectric continuum with a relative permittivity epsilon = 2. The time histories of the individual angles between NH bond vectors at time 0 and time t later are calculated, and then Gaussian smoothing of typically 50 ps is applied. The smoothed data are subtracted from the original data to yield the short time fluctuations of the NH bond vectors, and then the rms deviations of the angles are calculated and compared to experimental NMR results. Pf1 coat protein and the c subunit of the E. coli F1F0 ATP synthase are used as examples of membrane proteins. The calculated NH bond rms fluctuations are in qualitative agreement with experimental NMR data in showing that each of these proteins has a mobile segment connecting two helices, as well as mobile N and C-terminal regions. This MD simulations protocol can demonstrate the presence of both the amphipathic and hydrophobic helices while hydropathy plots are able to detect only the hydrophobic helices present in membrane proteins. PMID- 7473723 TI - Transcriptional regulation of PS-IAA4/5 and PS-IAA6 early gene expression by indoleacetic acid and protein synthesis inhibitors in pea (Pisum sativum). AB - The transcription of two genes, PS-IAA4/5 and PS-IAA6, in pea is induced by indoleacetic acid (IAA) and protein synthesis inhibitors such as cycloheximide (CHX) and anisomycin (ANI). Induction by IAA is rapid, taking 5 and 7.5 minutes for PS-IAA4/5 and PS-IAA6, respectively, and is independent of IAA concentration and whether IAA has a free or esterified carboxyl group (ethyl-IAA). The rate of mRNA accumulation, however, is dependent on hormone concentration, and is greater with IAA than with ethyl-IAA. The turnover rates (t1/2) of the PS-IAA4/5 and PS IAA6 mRNAs are 60 and 75 minutes, respectively, and are not affected by IAA. CHX or ANI induce the transcription of PS-IAA4/5 and PS-IAA6 more slowly than IAA (5 to 10 minutes for PS-IAA4/5 and 20 minutes for PS-IAA6). While protein synthesis inhibitors stabilize both mRNAs, the rapidity of induction by CHX and ANI cannot be accounted for solely by mRNA stabilization. The relationship between mRNA induction and protein synthesis inhibition does not obey Michaelis-Menten kinetics, but rather is best described by a hyperbolic curve, suggesting the release of transcriptional repression by the inhibition of protein synthesis. RNA expression experiments with transgenic tobacco seedlings or with transfected pea protoplasts using PS-IAA4/5 promoter GUS or CAT fusions reveal that CHX transcriptionally activates PS-IAA4/5 gene expression. Thus, protein synthesis inhibitors have a dual effect on PS-IAA4/5 and PS-IAA6. (1) They stabilize both mRNAs (possibly by a translational arrest-linked process or by preventing the synthesis of a labile nuclease(s)). (2) They activate transcription (possibly by preventing the synthesis or function of a repressor). PMID- 7473724 TI - The non-inducible nature of super-repressors of the gal operon in Escherichia coli. AB - We isolated and characterized mutant repressors (GalR) of the gal operon in Escherichia coli. These repressors (super-repressors), called GalRs, have a non inducible phenotype. Repression of the gal operon by super-repressors cannot be lifted by inducer. The mutant galR genes, galRs, have been cloned and the mutational changes determined. Two of them, galRuv7s and galR78s, were located in the proposed sugar binding domains of the repressor. The repressor from wild-type (galR+), as well as from mutant galRuv7s, was purified and characterized biochemically. The results showed that, like wild-type GalR+, GalRuv7s binds to DNA normally and represses transcription from the P1 promoter and stimulates that from the P2 promoter of the gal operon. Nevertheless, compared to GalR+, GalRuv7s is much less sensitive to the presence of the inducer, D-galactose. The affinity of D-galactose to GalRuv7s is 10 to 30-fold lower, as measured by the effect of the inducer on GalR tryptophan fluorescence; GalR complexes with DNA and on GalR repression of transcription. Our results suggest that the super-repressor phenotype of GalRuv7s is because of a defect in D-galactose binding rather than a defect in the ligand-induced allosteric change or increased affinity for the operator. PMID- 7473726 TI - Energetics of coupled twist and writhe changes in closed circular pSM1 DNA. AB - The extent of local denaturation in closed circular pSM1 DNA depends upon the linking difference, delta Lk, and the temperature, t. We have determined the denaturation profiles, using gel electrophoresis, over the ranges -37 < or = delta Lk < or = +16 and 25 degrees C < or = t < or = 65 degrees C. We have applied statistical mechanical methods to these data to evaluate the free energies of superhelix formation, of the twisting of single strands around each other, and of the initration of local denaturation. Because the complete nucleotide sequence is needed for this analysis, the complete pSM1 DNA sequence was determined and is reported here. The values of the free energy parameters found in this work agree closely with those previously obtained from experiments with pBR322 DNA, suggesting that there is little dependence of these values on the particular DNA sequence. We find the temperature dependence of these free energies by the appropriate statistical mechanical analysis of the temperature dependent denaturation profiles produced by supercoiling. Calculations of the transition probability profiles indicate that the course of local denaturation in pSM1 DNA involves a complex competition among several sites of comparable susceptibility. This contrasts with the melting of pBR322 DNA, in which one principal site dominates. In both molecules the sites of predicted denaturation occur at or near regulatory regions, suggesting that duplex destabilization may be associated with their biological activities. PMID- 7473727 TI - Structure, thermodynamics and cooperativity of the glucocorticoid receptor DNA binding domain in complex with different response elements. Molecular dynamics simulation and free energy perturbation studies. AB - Molecular dynamics simulations and free energy perturbation calculations have been performed on the glucocorticoid receptor DNA-binding domain (GRDBD) in complex with three different half-site response elements (RE). These are the glucocorticoid receptor RE (GREH), the estrogen receptor RE (EREH) and an "intermediate" RE (GRE2H), where the base-pair 6A.T in GREH has been replaced by 6G.C. The molecular dynamics/free energy perturbation calculations reveal the same order of stability of the three complexes, i.e. GREH-GRDBD > GRE2H-GRDBD > EREH-GRDBD, as found experimentally, with reasonable quantitative agreement of the relative stabilities with the experiments. The more favourable entropy in associating GRDBD to GREH than to GRE2H could be explained by the additional non polar surface that is removed from the solvent in the former complex formation. Part of this additional surface originates from the 6T methyl group in GREH. Sequence-dependent hydration patterns of the DNA major groove, which is interrupted by the GRDBD binding, could also contribute to the relatively more favourable entropy in forming GREH-GRDBD than GRE2H-GRDBD. The tenfold reduced cooperativity in the dimeric GRDBD binding to a partially palindromic ERE (palERE) compared to palGRE/GRE2 could be attributed to conformational changes in the residues of the "D-box" (Cys476 to Cys482) observed in the molecular dynamics simulations of EREH-GRDBD, but not in the other two complexes. Moreover, the conformations of the base-pairs at positions 5 and 6 are distorted (compared to free B-DNA) in GREH and GRE2H, due to the binding of GRDBD. Since the corresponding conformational distortions were not found in EREH-GRDBD, it is plausible that the distortions mediate cooperative binding through a propagation of the distortions to the other half site, thus facilitating binding of the second GRDBD. PMID- 7473725 TI - Localization of sequences required for size-specific splicing of a small Drosophila intron in vitro. AB - Many introns in Drosophila and other invertebrates are less than 80 nucleotides in length, too small to be recognized by the vertebrate splicing machinery. Comparison of nuclear splicing extracts from human HeLa and Drosophila Kc cells has revealed species-specificity, consistent with the observed size differences. Here we present additional results with the 68 nucleotide fifth intron of the Drosophila myosin heavy chain gene. As observed with the 74 nucleotide second intron of the Drosophila white gene, the wild-type myosin intron is accurately spliced in a homologous extract, and increasing the size by 16 nucleotides both eliminates splicing in the Drosophila extract and allows accurate splicing in the human extract. In contrast to previous results, however, an upstream cryptic 5' splice site is activated when the wild-type myosin intron is tested in a human HeLa cell nuclear extract, resulting in the removal of a 98 nucleotide intron. The size dependence of splicing in Drosophila extracts is also intron-specific; we noted that a naturally larger (150 nucleotide) intron from the ftz gene is efficiently spliced in Kc cell extracts that do not splice enlarged introns (of 84, 90, 150 or 350 nucleotides) derived from the 74 nucleotide white intron. Here, we have exploited that observation, using a series of hybrid introns to show that a region of 46 nucleotides at the 3' end of the white intron is sufficient to confer the species-specific size effect. At least two sequence elements within this region, yet distinct from previously described branchpoint and pyrimidine tract signals, are required for efficient splicing of small hybrid introns in vitro. PMID- 7473728 TI - Calculation of relative binding free energies and configurational entropies: a structural and thermodynamic analysis of the nature of non-polar binding of thrombin inhibitors based on hirudin55-65. AB - Free energy calculations were carried out on a series of exosite-binding inhibitors of thrombin. These inhibitors are based on the C-terminal fragment of hirudin and have the sequence Phe-Glu-Glu-IleH59-Pro-Glu-Glu-Tyr- Leu, where the superscript over Ile indicates its relative position in the natural sequence of hirudin. In this study, the effect of replacing IleH59 with ten other non-polar amino acids was examined. Three preferred interaction sites for methyl/methylene groups for the various XaaH59 side-chains in the complex were identified from conformational search calculations. The corresponding thermodynamic changes were determined using a combination of systematic search and energy minimization in a manner that locates the local minima in the system and in the process simultaneously builds up the partition function. The free energy, internal energy and entropic contributions are readily calculated from the partition function. Very good agreement in the resulting relative binding free energies was obtained between theory and experiment. The calculations allowed us to dissect out the enthalpic, entropic and solvation contributions to delta delta G. The contribution from desolvation was found to be relatively weak. The binding of these non-polar side-chains to thrombin is found to be driven mainly by favorable protein-ligand interactions rather than by the desire for non-polar groups to be desolvated. We also find that the configurational entropy contributes about 0.48 kcal/mol (0.81 kappa T) in average for each torsional angle "frozen" in binding. PMID- 7473729 TI - Disulfide mutants of barnase. I: Changes in stability and structure assessed by biophysical methods and X-ray crystallography. AB - In this series of papers, we examine the effects of introducing disulfide bonds on the properties, structure and thermodynamics of a small globular protein, barnase. Three mutants have been made, in each of which a single crosslink confers different properties. Two of the disulfide bonds, between residues 43 and 80 (43-80) and between residues 85 and 102 (85-102), stabilise the protein, relative to both wild-type and the corresponding (reduced) dithiol forms: 85-102 is more stable than predicted from the entropic destabilisation of the unfolded state; 43-80 is less stable than predicted. The third disulfide bond, between residues 70 and 92 (70-92) destabilises the protein relative to both wild-type and the corresponding dithiol form, implying significant disruption of the folded protein on formation of the disulfide bond. Crystal structures of the three mutant proteins have been solved. All three proteins have essentially the same fold as wild-type, but with left-handed disulfide bonds, which have dihedral geometries that have not been observed in naturally occurring disulfides. In the very stable mutant 85-102, there is no significant difference between the mutant and wild-type structures: these data do not explain the large stability of this protein. The disulfide bond at 43-80 induces small structural rearrangements close to the site of the disulfide bond, associated with some local disorder: the crosslink appears to decrease the stability of the native form of the protein. The destabilising disulfide bond at 70-92 induces considerable structural change, with displacement of a loop and consequent disruption of a stabilising salt bridge. Our studies do not support the view that the conformation of the disulfide bond is crucial in determining the stability of the mutant proteins. PMID- 7473730 TI - Disulfide mutants of barnase. II: Changes in structure and local stability identified by hydrogen exchange. AB - The hydrogen exchange behaviour of two stabilised disulfide mutants of barnase has been examined using NMR H/2H exchange measurements. The choice of experimental conditions is crucial in experiments to study the effects of mutations on local and global stability and dynamic behaviour of proteins. If exchange conditions allow both local and global unfolding events to be examined, then a comparison of three proteins (two mutants and wild-type) allows the effect of a mutation on the folded state to be ascribed to specific increases or decreases in local stability. This method was used to examine the effect of an introduced crosslink on the folded state of two different disulfide mutants of barnase, and the results are related to structural studies. It is found that disulfide bonds can stabilise or destabilise local structures as well as having an effect upon global stability. The effects of the mutations on exchange rate constants of protons are compared with the effects on the structure upon the introduction of the disulfide bonds. In the protein with a disulfide bond between residues 43 and 80, some local exchange rate constants are higher, reflecting local destabilisation at the site of the disulfide bond, associated with small structural rearrangements. In the protein with a disulfide bond between residues 85 and 102, amide protons of the adjacent loop are protected to a considerable extent. This is not associated with a structural rearrangement yet indicates that this disulfide bond has an effect on the stability of this loop. PMID- 7473731 TI - Helix-destabilizing activity of phi 29 single-stranded DNA binding protein: effect on the elongation rate during strand displacement DNA replication. AB - The single-stranded DNA (ssDNA)-binding protein (SSB) of bacteriophage phi 29 is one of the virus-encoded proteins required for viral DNA replication. We have found that phi 29 SSB has helix-destabilizing activity since it removes secondary structure of the ssDNA in phi 29 replicative intermediates, as revealed by electron microscopy, and displaces oligonucleotides annealed to M13 ssDNA. To investigate the mechanism of the SSB-dependent stimulation of phi 29 DNA replication we have characterized the helix-destabilizing activity of phi 29 SSB and measured its effect on the DNA elongation rate by phi 29 DNA polymerase, which does not require an accessory helicase. The use of replication reactions where strand displacement is either required (phi 29 DNA replication) or not (conversion of primed M13 ssDNA into double-stranded DNA (dsDNA)) has allowed us to find that (1) strand displacement DNA replication was affected by lowering the temperature or by increasing the salt concentration, since the DNA elongation rate on the phi 29 template was three to fourfold slower than on primed M13 ssDNA, (2) under those conditions, addition of phi 29 SSB stimulated to different extents the DNA elongation rate during phi 29 DNA replication, whereas it had a marginal effect on primed M13 ssDNA replication, and (3) phi 29 SSB increased four to sixfold the phi 29 DNA elongation rate by phi 29 DNA polymerase strand displacement mutants, reaching approximately 50% the rate of the wild-type enzyme. The implications of the helix-destabilizing properties of the phi 29 SSB under conditions in which DNA opening is impaired are discussed. PMID- 7473732 TI - A model for transmembrane signalling by the aspartate receptor based on random cassette mutagenesis and site-directed disulfide cross-linking. AB - Extracellular information is transduced by transmembrane receptors into the inside of the cell across a membrane barrier. To understand the molecular basis of transmembrane signalling, we replaced the transmembrane segment 2 (TM2) of the Escherichia coli aspartate receptor, Tar, with random sequences that are 21 amino acid residues in length and consist of Arg, Gly, Ser, Cys, Val, Leu, Ile and Phe at each position. From this ensemble for recombinant molecules, functional receptors were recovered as clones that could bind aspartate and transmit a signal to the intracellular domain. Restricted average hydrophobicity values were observed for functional transmembrane domains, and support the observation that transmembrane segments typically have hydrophobicity values greater than 1.6. However, non-functional transmembrane domains with greater hydrophobicity than 1.6 indicate that hydrophobicity is not a sole determinant for its function. Fourier transform analysis of the functional TM2 sequences suggests that the transmembrane segment has an alpha-helical structure with three distinct faces. Cross-linking of the faces to transmembrane segment 1 (TM1) mimics the "locked" signalling phenotypes of the wild-type receptor. The results are consistent with a model in which TM2 rotates in the plane of the lipid bilayer, and the rotation becomes locked at one face of the alpha-helix in the presence of attractant and at another face in the presence of repellent. This dynamic movement of the transmembrane domain may be a common signalling mechanism of homologous membrane receptor molecules such as the insulin receptor. Random-cassette mutagenesis and disulfide cross-linking provide powerful strategies for examining the structure and function of transmembrane segments. PMID- 7473733 TI - Radial mass analysis of the flagellar filament of Salmonella: implications for the subunit folding. AB - X-ray fiber diffraction patterns of the R-type straight flagellar filament of Salmonella typhimurium SJW1655 strain showed layer-lines with an axial spacing of 1/437 A-1, which could be resolved only due to very small disorientation angles (< 2 degrees) of the filaments in oriented sol specimens. Although the equatorial layer-line was situated between the relatively strong first layer-lines right above and below it, these small disorientation angles and a new method of two dimensional angular deconvolution allowed us to determine the equatorial layer line intensities quite accurately. The equatorial data were phased by using the amplitude difference between the native flagellar filament and its heavy atom derivatives. One of the heavy-atom derivatives was prepared by introducing a cysteine residue by site-directed mutagenesis and applying a mercury compound. From the equatorial structure factors, the radial density distribution of the filament was calculated at 11 A resolution. A prominent feature was two pairs of high density peaks at radii of around 25 and 45 A and a deep density trough between them, which corresponds to the concentric double tubular structure in the core region that has been found in the density map recently deduced by helical image reconstruction from electron micrographs of frozen hydrated filaments. The molecular masses were estimated for four radial segments that correspond to the morphological domains identified in the map of helical image reconstruction. Then the domains were assigned to sequence positions by correlating the estimated masses with those of proteolytic fragments of flagellin. The assignment is consistent with the distributions of secondary structures and in particular alpha helical coiled-coils that were predicted from the sequence. It also helps to understand how the polymerization behaviour is affected by truncation of the disordered terminal regions of flagellin and why mutations in a specific region are responsible for changes in the polymorphic shape of the filament. PMID- 7473734 TI - The crystal structure of the antibody N10-staphylococcal nuclease complex at 2.9 A resolution. AB - The three-dimensional structure of the antibody N10 Fab fragment complexed with staphylococcal nuclease (SNase) has been determined to 2.9 A resolution. Eighteen residues from six complementarity-determining regions (CDR) recognize an epitope of five distinct SNase segments with a total of 17 residues. The overall shape of the antibody-antigen interface is U-shaped rather than the more or less rectangular interface seen in other antibody-protein antigen interfaces. Despite the U-shaped interface, the amount of surface buried in the complex, 828 A2 for SNase and 793 A2 for N10, is typical of antibody-protein antigen complexes. Contributing to the shape of the interface is the shortest antibody heavy chain CDR3 loop reported to date, which probably allows access of bulk solvent in the center of the "U" interface. Another unusual feature of the N10 antibody is the 15 residue antibody light chain-CDR1, a length seen in only three other reported antibodies. Antibody light chain-CDR1 displays a previously unobserved conformation in its distal portion. Finally, although some of the movement observed in the antibody-bound SNase may be due to crystal contacts, it is clear that some side-chain rearrangements are the result of antigen-antibody interaction. PMID- 7473735 TI - Mechanism of hydrogen-deuterium exchange in trp repressor studied by 1H-15N NMR. AB - Amide proton exchange rates have been measured for fast-exchanging amides in trp aporepressor, and compared with the rates measured in the holorepressor. The results indicate that the presence of the ligand stabilizes all of the amide protons in the molecule against exchange, not just those whose access to solvent it directly hinders. This global hindering of the exchange process by tryptophan implies that there is a non-random element in the transmission mechanism, so that damping of the exchange in one part of the molecule also damps exchange in another region. This damping at a distance is not associated with any measurable changes in the intervening average secondary structure. This suggests the existence of a concerted dynamic process in the protein backbone that is modulated by ligand binding and in turn affects the observed backbone proton exchange. PMID- 7473736 TI - Determination of the three-dimensional solution structure of scyllatoxin by 1H nuclear magnetic resonance. AB - The three-dimensional solution structure of Scyllatoxin (leiurotoxin I) a venom peptide from the scorpion Leiurus quinquestriatus hebraeus was determined at 1 A resolution by homonuclear proton n.m.r. methods at 500 MHz. Data analysis and structure calculation followed conventional protocols inherent to DIANA and related programs with two exceptions. First, distance constraints were obtained from two-dimensional nuclear Overhauser spectra by a previously described partial relaxation matrix approach. Second, since the pairing pattern of the six cysteine residues was not established a priori, the unequivocal assignment of the disulfide bridges was achieved exclusively from the n.m.r. data by a statistical analysis of preliminary DIANA structures. In the final calculation we used 227 upper distance constraints, 67 torsional constraints from vicinal coupling constants and ten stereospecific assignments of beta-methylene protons. Scyllatoxin folds into a compact ellipsoidal shape. An alpha-helix (defined with 0.24 A resolution) spanning 2.5 turns from Leu5 till Ser14 is stabilized by Cys8 Cys26 and Cys12-Cys28 disulfide bridges to the carboxy-terminal strand of an anti parallel beta-sheet. The antiparallel beta-sheet (defined at 0.66 A resolution) extends from Leu18 to Val29 with a tight turn at Gly23-Asp24 and displays a right handed twist. Scyllatoxin competes with the toxins apamin from Apis mellifera mellifera and P05 from Androctonus mauretanicus mauretanicus for the same or similar high conductance calcium-activated potassium channels. Consideration of presently known biological activities and three-dimensional structures of these toxins suggest a different toxin-receptor interaction of scyllatoxin as compared to apamin and P05. PMID- 7473737 TI - Mutations at positions 153 and 328 in Escherichia coli alkaline phosphatase provide insight towards the structure and function of mammalian and yeast alkaline phosphatases. AB - In order to understand some of the differences between human placental, human, Saccharomyces cerevisiae and Escherichia coli alkaline phosphatases in specific activity, activation by magnesium, and pH versus activity profiles, the X-ray crystal structures of three mutant E. coli alkaline phosphatases have been determined. The aligned sequences of alkaline phosphatases from mammalian, yeast and E. coli show that 25 to 30% of the amino acids are absolutely conserved and the active site residues are completely conserved with the exception of residues 153, 328 and 155. The bacterial enzyme has a salt-bridge, Asp153/Lys328, near the third metal binding site which, based on sequence homology, is apparently absent in the yeast and mammalian enzymes. The human enzymes have histidine at positions 153 and 328, and the yeast enzyme has histidine at position 328. In the E. coli enzyme, Asp153 was replaced by histidine (D153H), Lys328 was replaced by histidine (K328H), and a double mutant (DM) was constructed containing both mutations. The structure of the K328H enzyme was refined using cross-validation to a resolution of 2.3 A with a working R-factor of 0.181 and a free R-factor of 0.249. The DM structure was determined to a resolution of 2.5 A with a working R factor of 0.166 and a free R-factor of 0.233. The structure of the D135H enzyme, which has been reported to a resolution of 2.4 A, has been re-refined using cross validation to a working R-factor of 0.179 and a free R-factor of 0.239 for controlled comparisons with the two new structures. In all three structures the most significant changes are related to the bound phosphate inhibitor and the identity of the metal ion in the third binding site. The changes in the position of the phosphate group and the alterations at the third metal binding site indicate the structural basis for the variations in the steady-state kinetic parameters previously reported for these enzymes. PMID- 7473738 TI - Structure-guided analysis reveals nine sequence motifs conserved among DNA amino methyltransferases, and suggests a catalytic mechanism for these enzymes. AB - Previous X-ray crystallographic studies have revealed that the catalytic domain of a DNA methyltransferase (Mtase) generating C5-methylcytosine bears a striking structural similarity to that of a Mtase generating N6-methyladenine. Guided by this common structure, we performed a multiple sequence alignment of 42 amino Mtases (N6-adenine and N4-cytosine). This comparison revealed nine conserved motifs, corresponding to the motifs I to VIII and X previously defined in C5 cytosine Mtases. The amino and C5-cytosine Mtases thus appear to be more closely related than has been appreciated. The amino Mtases could be divided into three groups, based on the sequential order of motifs, and this variation in order may explain why only two motifs were previously recognized in the amino Mtases. The Mtases grouped in this way show several other group-specific properties, including differences in amino acid sequence, molecular mass and DNA sequence specificity. Surprisingly, the N4-cytosine and N6-adenine Mtases do not form separate groups. These results have implications for the catalytic mechanisms, evolution and diversification of this family of enzymes. Furthermore, a comparative analysis of the S-adenosyl-L-methionine and adenine/cytosine binding pockets suggests that, structurally and functionally, they are remarkably similar to one another. PMID- 7473739 TI - Automatic analysis of protein conformational changes by multiple linkage clustering. AB - An automatic algorithm is presented for analyzing protein conformational changes such as those occurring upon substrate binding or in different crystal forms of the same protein. Using, as sole information, the atomic coordinates of a pair of protein structures, the procedure first generates structure alignments, which optimize the root-mean-square deviation of the backbone atoms. To this end, equivalent secondary structures and/or loops from both proteins are combined by a multiple linkage hierarchic clustering algorithm, which generates several intertwined clustering trees. Automatic analysis of these clustering trees is used to dissect the mechanism of the conformational change. It allows the identification of the static core, representing the collection of secondary structures which undergo no structural changes, as well as other entities which move like rigid bodies. It also permits the description of the movement of secondary structures or loops relative to this core or entities. USing this information, it can be inferred whether a particular conformational change involves shear or hinge motion, or components of both. The algorithm is applied to the analysis of the conformational changes of citrate synthase, lactate dehydrogenase, lactoferrin and beta-glucosyltransferase, representing typical examples of shear- and hinge-type mechanisms, and a varied range in movement size. The results are shown to be in excellent agreement with previous analyses, and to provide additional information which gives a more complete and objective picture of the conformational change. Using our automatic algorithm, we find that any conformational change may be viewed as having components of both shear- and hinge-type motion. Determining which of these is most appropriate requires the combination of the information provided by our procedure with detailed knowledge of the protein tertiary structures. PMID- 7473740 TI - Different subdomains are most protected from hydrogen exchange in the molten globule and native states of human alpha-lactalbumin. AB - alpha-Lactalbumin (alpha-LA) is a two-domain, calcium-binding protein that forms one of the best studied molten globules. We present here amide hydrogen exchange studies of the molten globule formed by human alpha-LA at pH 2 and compare these results with a similar study of the native state at pH 6.3. The most persistent structure in the molten globule is localized in the helical domain, consistent with previous results. However, the helices most protected from hydrogen exchange in the molten globule are, in the native state, less protected from exchange than other regions of the protein. The molten globule appears to contain major elements of the native fold, but formation of the fully native state requires stabilization of structure around the calcium-binding site and domain interface. PMID- 7473741 TI - Site-directed mutagenesis of proteolytic antibody light chain. AB - Amino acid residues in a proteolytic antibody light chain selected by molecular modeling were substituted with Ala by site-directed mutagenesis. Hydrolysis of vasoactive intestinal polypeptide (VIP), the immunogen employed to elicit the antibody light chain, was reduced by > 95% by replacement of Ser27a or His93 by Ala residues. Similar reductions in the activity were observed using synthetic protease substrates containing Arg-methylcoumarinamide (MCA) and Lys-MCA bonds. Turnover of the Ser27a and His93 mutants was lower than that of wild-type protein by about two orders of magnitude. The activity of the wild-type protein was inhibited selectively by diisopropylfluorophosphate (DFP), a serine protease inhibitor, but the residual activity of the Ser26 mutant was refractory to DFP. The affinity of the wild-type light chain for the substrate ground state was nearly unaffected by mutations at Ser27a and His93. In contrast, a Ser26 single mutant and a His27d/Asp28 double mutant displayed increased Km (by about tenfold) and increased turnover (by about tenfold) using VIP as substrate. The kinetic constants for these mutants and the wild type protein were essentially identical with Boc-Glu-Ala-Arg as substrate. Thus, two types of residues participating in catalysis by the light chain have been identified. Ser27a and His93 are essential for catalysis but not for initial high affinity complexation and substrate. Ser26 and His27d or Asp28 participate in VIP binding and limit turnover indirectly. PMID- 7473742 TI - Activity of a C. elegans GATA transcription factor, ELT-1, expressed in yeast. AB - The GATA motif (WGATAR) is found in the promoter regions of numerous Caenorhabditis elegans genes, including two intestine-specific genes, vit-2 and ges-1, in which it has been shown to be required for promoter function. The protein ELT-1, encoded by a single-copy gene homologous to the GATA family of vertebrate transcription factors, is potentially capable of interacting with this element. In order to determine whether ELT-1 is a transcriptional activator that recognizes this sequence, we have expressed it under the control of the GAL1 promoter in yeast. lacZ driven by the CYC1 promoter lacking an upstream activation sequence (UAS) but containing GATA sequences was used as a reporter. beta-Galactosidase was expressed upon induction only when GATA sequences were present, and expression was increased dramatically by additional binding sites. Deletion analysis demonstrated that the C terminus, containing only one of the two zinc fingers, is sufficient for activation. In addition, the DNA-binding domain and two transactivation regions were identified by fusing these isolated domains to previously defined domains of heterologous transcription factors. While most single base alterations in the GATA core sequence eliminated activity, an A to C change in position four, creating a GATC core, was found to increase activity significantly. The deleted ELT-1 protein containing only the C-terminal Zn finger was sufficient for activation in response to GATA, but both fingers were required for activation at GATC. A variety of sites with non-optimal sequences surrounding the GATA core also were found to be excluded better by the protein containing both Zn fingers. Furthermore, a fusion protein containing the entire ELT-1 DNA binding domain fused to the VP16 activation domain was found to have an even greater preference for the GATC core, as well as the optimal flanking bases. We conclude that, although ELT-1 having only its C-terminal finger is capable of activation in response to the WGATAR site, the presence of the upstream finger supplies additional base specificity. PMID- 7473743 TI - Functional redundancy of promoter elements ensures efficient transcription of the human 7SK gene in vivo. AB - Deletion and mutation studies of the human 7SK gene transfected into HeLa cells have identified three functional regions of the promoter corresponding to the TATA box at -25, the proximal sequence element (PSE) between -49 and -65 and the distal sequence element (DSE) between -243 and -210. These elements show sequence homology to equivalent regions in other snRNA genes and are functionally analogous. Unlike the DSEs of many snRNA genes however, the 7SK DSE does not contain a consensus binding site for the transcription factor Oct-1 but rather, contains two non-consensus Oct-1 binding sites that can function independently of one another to enhance transcription. Unusually, the 7SK PSE can retain function even after extensive mutation and removal of the conserved TGACC of the PSE has little effect in the context of the whole promoter. However, the same mutation abolishes transcription in the absence of the DSE suggesting that protein/protein interactions between DSE and PSE binding factors can compensate for a mutant PSE. Mutation of the 7SK TATA box allows snRNA type transcription by RNA polymerase II to occur and this is enhanced by the DSE, indicating that both the DSE and the PSE can also function with pol II. In addition, mutation of the TATA box does not abolish pol III dependent transcription, suggesting that other sequence elements may also play a role in the determination of polymerase specificity. Although the human 7SK gene is transcribed efficiently in Xenopus oocytes, analysis of the 7SK wild-type gene and mutants in Xenopus oocytes gives significantly different results from the analysis in HeLa cells indicating that the recognition of functional elements is not the same in the two systems. PMID- 7473744 TI - The sixty nucleotide OccR operator contains a subsite essential and sufficient for OccR binding and a second subsite required for ligand-responsive DNA bending. AB - OccR is a transcriptional regulatory protein of Agrobacterium tumefaciens that activates the occQ operon in response to octopine, an arginine derivative released from plant tumors. OccR binds to its operator with similar affinity and the same stoichiometry in the presence or absence of octopine, but octopine shortens the protein's DNase I footprint and partially relaxes an OccR-incited DNA bend. In this study, resections and other alterations of the operator were used to demonstrate that 19 nucleotides near the end of the operator furthest from the occQ promoter were essential for high affinity OccR binding. This sequence, denoted the high affinity subsite, was sufficient for binding, provided that the deleted operator sequences were replaced with vector-derived DNA. The same number of OccR monomers bound to resected operators as to the wild-type operator, and OccR was able to protect vector-derived sequences adjacent to the high affinity subsite. Sequences at the promoter proximal end of the operator were required for wild-type patterns of ligand-responsive DNA bending. A sequence alteration at the end furthest from the high affinity subsite caused a partially locked low angle DNA bend, while two more centrally localized mutations caused fully or partially locked high angle bends. This suggests that the promoter proximal half of the operator may contain at least two sites required for wild type ligand-responsive DNA bending. These mutations also provided evidence that the partial relaxation of this bend by octopine may be essential for occQ activation. PMID- 7473745 TI - Genetic dissection of intersubunit contacts within human protein kinase CK2. AB - The activity of the catalytic alpha subunits of protein kinase CK2 is modulated by interaction with the regulatory beta subunits. In order to define the domains involved in intersubunit contacts, we have applied the two-hybrid system, which a is yeast-based genetic method for the detection of protein-protein interactions in vivo. The data demonstrate that the alpha and beta subunits interact with each other and that the beta subunits, but not the alpha subunits, are able to self associate. This suggests that the beta subunits play a bridging role in the architecture of the CK2 holoenzyme by linking two alpha:beta heterodimers into a tetrameric complex. Analysis of truncated alpha and beta subunits was used to delimit the subregions necessary for complex formation. The data reveal that the beta subunit is modular in structure, with the two fully separable domains involved in homomeric beta:beta and heteromeric alpha:beta interactions, respectively. Also, beta subunits lacking the autophosphorylation sites in the N termini are able to associate with both the alpha and beta subunits. Furthermore, we find that the N terminus and the evolutionarily less conserved C terminus of the alpha subunit are dispensable for establishing heterodimeric alpha:beta structures. PMID- 7473746 TI - The 60 kDa heat shock proteins in the hyperthermophilic archaeon Sulfolobus shibatae. AB - One of the most abundant proteins in the hyperthermophilic archaeon Sulfolobus shibatae is the 59 kDa heat shock protein (TF55) that is believed to form a homo oligomeric double ring complex structurally similar to the bacterial chaperonins. We discovered a second protein subunit in the S. shibatae ring complex (referred to as alpha) that is stoichiometric with TF55 (renamed beta). The gene and flanking regions of alpha were cloned and sequenced and its inferred amino acid sequence has 54.4% identity and 74.4% similarity to beta. Transcription start sites for both alpha and beta were mapped and three potential transcription regulatory regions were identified. Northern analyses of cultures shifted from normal growth temperatures (70 to 75 degrees C) to heat shock temperatures (85 to 90 degrees C) indicated that the levels of alpha and beta mRNAs increased during heat shock, but at all temperatures their relative proportions remained constant. Monitoring protein synthesis by autoradiography of total proteins from cultures pulse labeled with L(-)[35S]methionine at normal and heat shock temperatures indicated significant increases in alpha and beta synthesis during heat shock. Under extreme heat shock conditions (> or = 90 degrees C) alpha and beta appeared to be the only two proteins synthesized. The purified alpha and beta subunits combined to form high molecular mass complexes with similar mobilities on native polyacrylamide gels to the complexes isolated directly from cells. Equal proportions of the two subunits gave the greatest yield of the complex, which we refer to as a "rosettasome". It is argued that the rosettasome consists of two homo-oligomeric rings; one of alpha and the other of beta. Polyclonal antibodies against alpha and beta from S. shibatae cross-reacted with proteins of similar molecular mass in 10 out of the 17 archaeal species tested, suggesting that the two rosettasome proteins are highly conserved among the archaea. The archaeal sequences were aligned with bacterial and eukaryotic chaperonins to generate a phylogenetic tree. The tree reveals the close relationship between the archaeal rosettasomes and the eukaryotic TCP1 protein family and the distant relationship to the bacterial GroEL/HSP60 proteins. PMID- 7473747 TI - A double mutation in subunit c of the Na(+)-specific F1F0-ATPase of Propionigenium modestum results in a switch from Na+ to H(+)-coupled ATP synthesis in the Escherichia coli host cells. AB - The in vivo synthesis of an F1F0-ATPase hybrid in Escherichia coli strain PEF42 which harbours the genes for the Propionigenium modestum subunits a, b, c, and delta, a gene for hybrid alpha subunit with the N-terminal portion (amino acids 1 to 173) of P. modestum and the C-terminal region (amino acids 176 to 513) from E. coli, and the genes for the E. coli subunits beta, gamma and epsilon, yielded a functional enzyme complex. This hybrid ATPase coupled ATP synthesis to Na+ transport and required Na+ for growth on succinate. After random mutagenesis of the P. modestum genes of strain PEF42, clones were selected that grew on succinate in the absence of Na+. A double-mutation cPhe84Leu, cLeu87Val that was found in several of these clones, was introduced by site specific mutagenesis into the parent strain PEF42. The resulting strain E. coli MPC8487 also exhibited Na(+)-independent growth on succinate, showing that the double mutation is the only reason for the new phenotype. The mutation causes a change of the coupling ions of the hybrid ATPase from Na+ in strain PEF42 to H+ in strain MPC8487. This conclusion was supported by the biochemical properties of the ATPase from strain MPC8487. Unlike the parent enzyme, the mutated ATPase was not activated by Na+, but retained activation by Li+. The pH optimum of the mutated ATPase (in the absence of Na+ or Li+) was shifted from pH 6.5 to pH 7.5, and the specific ATPase activity of the cell membranes increased about fourfold over that found in membranes of the parent cells. The mutated ATPase pumped protons or Li+ after reconstitution into proteoliposomes, and the transport of both cations was not affected by Na+. The double mutation in the c subunit thus results in the loss of Na+ binding, retention of Li+ binding and an improvement of H+ binding. PMID- 7473748 TI - Autocatalytic gene expression occurs via transertion and membrane domain formation and underlies differentiation in bacteria: a model. AB - When bacteria contain two chromosomes, two or more copies of the same gene are present in the same cytoplasm and, if these copies are subject to negative regulation in trans and positive (autocatalytic) regulation in cis, one copy will be expressed at the expense of the other copy(ies). This autocatalytic process depends on the coupled transcription, or translation and insertion of nascent proteins into the membrane, or transertion. Transertion is responsible for looping genes out of the nucleoid and increasing their accessibility to transcription factors. Transertion of proteins with lipid preferences creates proteolipid domains in the membrane. These domains fuse to give two types of large domains, each associated with the expression of a particular set of genes. These large domains organize kinases, proteases and transcription factors and result in the expression of one set of genes encoding proteins with common lipid preferences from one chromosome and expression of a different set from the other. These intracellular differences underlie the production of different progeny by cell division that follows, for example, reception of extracellular signals, and that constitutes differentiation in bacteria. PMID- 7473749 TI - Three-dimensional solution structure of bombyxin-II an insulin-like peptide of the silkmoth Bombyx mori: structural comparison with insulin and relaxin. AB - The three-dimensional solution structure of bombyxin-II, an insulin-like two chain peptide produced by the brain of the silkworm Bombyx mori, has been determined by simulated annealing calculations based on 535 distance constraints and 24 torsion-angle constraints derived from NMR data and three distance constraints of the disulfide bonds. To our knowledge, this is the first three dimensional structure determined for an invertebrate insulin-related peptide. The root-mean-square deviations between the best 10 structures and the mean structure are 0.58(+/- 0.15) A for the backbone heavy atoms (N, C alpha, C) and 1.03(+/- 0.18) A for all non-hydrogen atom if less well-defined N and C termini (A1, A20, B(-2) to B4 and B23 to B25) are excluded. The overall main-chain structure of bombyxin-II is similar to that of insulin. However, there are significant conformational and functional differences in their B-chain C-terminal parts. The B-chain C-terminal part of bombyxin-II adopts an extension of the B-chain central helix like that of relaxin and is not required for bombyxin activity, while the corresponding part of insulin adopts a sharp turn and a beta-strand and is essential for insulin activity. This structure demonstrates that bombyxin-II is more closely related to relaxin than to insulin, and suggests that insulin might have evolved the additional receptor-recognition site in the B-chain C-terminal beta-strand to distinguish itself from bombyxin and relaxin. The structure of bombyxin-II thus provides novel insights into the receptor recognition and divergent molecular evolution of insulin-superfamily peptides. PMID- 7473750 TI - Identification of the receptor-recognition surface of bombyxin-II, an insulin like peptide of the silkmoth Bombyx mori: critical importance of the B-chain central part. AB - Bombyxin-II, a brain-secretory peptide of the silkmoth Bombyx mori, shares 40% sequence identify and the characteristics core structure with human insulin. In spite of the structural similarity, no cross-activity is observed between them. To localize the active region of bombyxin-II, we have synthesized chimeric molecules of bombyxin-II and human insulin, and examined their bombyxin activity. Two chimeric molecules, which were sequentially identical except for the B-chain central part, showed significantly different potencies in bombyxin activity. Solution structure determination of these chimeric molecules revealed that their B-chain central parts took similar main-chain conformation, but formed dissimilar patches on their molecular surfaces. Therefore, the surface patch formed by the central part of the bombyxin-II B-chain is of critical importance for recognition of the bombyxin receptor. The above results, together with other data on the structure-activity relationships of bombyxin, indicate that the receptor recognition surface of bombyxin-II includes the A-chain N and C, termini in addition to the B-chain central part. Though bombyxin-II, human insulin and human relaxin 2 use the common surface as their receptor-recognition sites, each of the surface patches is characterized by the variety of involved side-chains. Insulin and relaxin involve additional parts for receptor recognition, particularly the B chain C-terminal part and the extended A-chain N-terminal helix, respectively. In conclusion, these ligands have evolved their own specific mechanisms for receptor recognition while retaining the major recognition surface. PMID- 7473751 TI - An integrated kinetic analysis of intermediates and transition states in protein folding reactions. AB - Relaxation rates for folding and unfolding of two proteins have been measured over a range of denaturant concentrations to examine the reaction pathways leading to the late transition state. The proteins were chosen for their marked differences in both kinetic and structural properties. Results for the N-terminal domain of phosophoglycerate kinase (N-PGK), from Bacillus stearothermophilus, reveal the existence of a single intermediate (pathway = U-I-F), and obey the general relationship: kobs = k(U-I) + k(I-U)/[1 + 1/KU/I]. Hen egg white lysozyme folds through two intermediates (pathway = U-I-I.-F) and the relaxation kinetics for formation and decay of the fully folded state are described by: kobs = k(F I.)+KI.-F)/[1+ 1/KI./I+ 1/(KI/U.KI./I)]. Rate constants apply to the first step in unfolding and the last step in folding, respectively, these being rate limiting in the stated directions. Equilibrium constants describe the stability of transient intermediates, as indicated by the subscripts. Rate constants alter with denaturant according to the generalized equation k = kw.exp((mg-mt).D), where kw is the rate constant in water, mg and mt are parameters describing the relative solvent exposures of the ground and transition state conformations respectively, and D is the calculated denaturant activity. The same principle applies to equilibrium constants for rapid steps, i.e. for a process A = B; KA/B = KA/B(w).exp((mB-mA).D). The combined application of these relationships allows measurement of the relative free energy and degree of solvation or compactness of intermediates and transition states in folding pathways from a single set of kinetic data. In the case of lysozyme, the fast but measurable rates of interconversion of intermediate states (I and I.) have been examined by use of a sequential mixing technique, so providing additional information on a transition state which is not rate-limiting in the overall pathway. The analysis of rate profiles for folding and unfolding of these proteins yields parameters which are in precise agreement with those derived from equilibrium data. PMID- 7473753 TI - Ionisation of cysteine residues at the termini of model alpha-helical peptides. Relevance to unusual thiol pKa values in proteins of the thioredoxin family. AB - The physical basis of the unusually low pKa values of an active site cysteine thiol group in proteins with the thioredoxin fold is unknown. The electrostatic field associated with an alpha-helix pointing with its N terminus towards the cysteine residue has been implicated to lower the thiol pKa value by up to 5 pH units in glutaredoxin and DsbA. Here, the influence of the presence of an alpha helical conformation on the ionisation of a cysteine thiol group located at or near the helix terminus is investigated in highly helical synthetic peptides with the generic sequence Ac-AAAAAAAAARAAAARAAAARAA-(NH2). The thiol pKa values have been determined by monitoring the pH dependence of the absorbance at 240 nm, of the alpha-helix content measured by the mean residue ellipticity at 222 nm, and of the chemical shifts of protons close to the sulphur atom of the cysteine residue. The favourable interaction between the thiolate anion at the N terminus and the alpha-helix decreases the thiol pKa value by up to 1.6 pH units when compared to a normal thiol pKa value measured in an unfolded control peptide, corresponding to a stabilisation energy of 2.1 kcal/mol. At the C terminus, the thiol pKa value is increased, but by only 0.2 pH units. The observations are consistent with an interaction of the alpha-helix dipole with the cysteine thiolate anion, involving both its charge and hydrogen-bonding. Subtle conformational effects in different model peptides appear to influence the ionisation of the thiol group significantly, with an N terminal Cys-Pro sequence having the most favourable interaction with the alpha-helix. PMID- 7473752 TI - Thermodynamic investigation of hirudin binding to the slow and fast forms of thrombin: evidence for folding transitions in the inhibitor and protease coupled to binding. AB - Temperature dependent studies of the interaction of the clotting enzyme thrombin with the potent natural inhibitor hirudin reveal a large negative heat capacity change of -1.7(+/- 0.2) kcal/mol per K associated with the formation of the thrombin-hirudin complex, independent of the allosteric state of the enzyme. Binding of N-terminal fragments of hirudin (hir1-49 and hir1-43) is characterized by heat capacity changes of -1.2(+/- 0.1) and -0.9(+/- 0.1) kcal/mol per K, respectively. The magnitude of these heat capacity changes is unprecedented for protease-inhibitor interactions. A thermodynamic analysis based on observed heat capacity and entropy changes predicts that binding is accompanied by substantial coupled folding transitions in both hirudin and thrombin. In the absence of a structure of free thrombin, analysis of differences in the predicted number of residues which fold upon binding hirudin and its fragments leads to the following structural model: three surface loops in thrombin (W60d, W148 and fibrinogen binding loops) are disordered in the free state and fold upon formation of the thrombin-hirudin complex. Molecular dynamics simulations, run over a time scale of 5 ps, are consistent with the hypothesis of large scale coupled folding transitions in both hirudin and thrombin upon formation of the complex. Comparison of the thermodynamics for the interaction of hirudin with the slow and fast forms of thrombin allows dissection of the coupling free energy for allosteric switching. The coupling free energy for the slow-->fast transition increases linearly, in absolute value, with temperature. The coupling enthalpy and entropy terms for hirudin were found to be delta Hoc = 12(+/- 1) kcal/mol and delta Soc = 47(+/- 4) cal/mol per K. Preferential interaction with the fast form is therefore due to the balance of two opposite forces, both quite large in magnitude. The contribution of enthalpic effects opposes the slow-->fast transition and stabilizes binding to the slow form. The contribution of entropic effects favors the slow-->fast transition and stabilizes binding to the fast form. In the physiological temperature range the entropic effects prevail and result in preferential binding of hirudin to the fast form. The region of thrombin recognizing the N-terminal domain of hirudin contains most of the residues that are energetically linked to the slow-->fast transition. This region is part of the "allosteric core" of thrombin and includes the W60d loop, shaping the specificity site S2, and the Na+ binding loop connecting the last two beta strands of the B chain. PMID- 7473754 TI - Identification in the human genome of mobile elements spread by DNA-mediated transposition. AB - We have identified in the human genome two families of mobile elements possessing the sequence characteristics of transposons that move directly from DNA to DNA rather than requiring the reverse transcription of an RNA intermediate. One type of element is closely related to the autonomous transposable element, mariner, and comprises a coding region for a transposase protein flanked by short terminal inverted repeat sequences (TIRs) of 31 or 32 bp. Elements of the second type form a family of short interspersed repetitive elements (SINEs) that are composed simply of two 37 bp TIRs surrounding six unique bps. The TIRs of the human mariner family are identical in all but one position to those of the SINE family, suggesting that the inverted-repeat SINEs represent non-autonomous transposable elements dependent on mariner-type transposase for mobility. Evidence for the mobility of both types of element is provided by examples of their integration into other repeat sequences and by the comparison of orthologous sites in cattle and human genomes. This evidence also shows that these elements have been active in DNA-mediated transposition at some point in the mammalian lineage. Therefore, it appears that the process of DNA-mediated transposition has occurred in mammalian cells and that its maximal cis-requirements are contained in the 80 bp consensus sequence of the human inverted-repeat SINE family. PMID- 7473755 TI - Dynamics of bacteriophage T4 DNA polymerase function: identification of amino acid residues that affect switching between polymerase and 3' --> 5' exonuclease activities. AB - Many DNA polymerases are multifunctional with the ability to replicate DNA as well as to proofread misincorporated nucleotides. Since polymerase and 3'--> 5' exonuclease activities appear to reside in spatially distinct active centers, there must be some mechanism for coordinating replication with proofreading and for transferring DNA between the two active centers. We have designed a genetic selection scheme to isolate bacteriophage T4 mutant DNA polymerases that are defective in "switching" between polymerase and exonuclease activities. Amino acid residues that affected active-site-switching were identified in four regions of the T4 DNA polymerase: two regions in the proposed exonuclease domain. Representative mutant DNA polymerases from each region were purified for biochemical studies. We propose that amino acid substitutions identified by mutational analysis affect critical contacts between T4 DNA polymerase and DNA that are required for transfer of DNA between the polymerase and exonuclease active centers. PMID- 7473756 TI - Anti-cruciform monoclonal antibody and cruciform DNA interaction. AB - Cruciform DNA structure, as a structural feature, has been associated with regulation of transcription, recombination and replication. Previously used to successfully modify DNA replication and affinity-purify origins and autonomously replicating sequences. Using enzyme protection assays, their binding activity has been localized to the base (elbow) of the cruciform stem. We report here the hydroxyl radical footprinting of 2D3 (kappa IgG1) anti-cruciform monoclonal antibody on a stable cruciform structure created by heteroduplexing fragments from two plasmids, identical except for two centrally located palindromes of different sequence. The footprinting was performed at near-physiological salt concentrations, conditions favouring the stacked X-structure of the cruciform. Our data show that binding by the antibody occurs at the four-way junction (elbows) of the stable cruciform. The binding of the antibody seems also to cause associated structural distortions in the heteroduplex, which generally result in greater sensitivity to hydroxyl radicals at the tips of the cruciforms. The data are consistent to hydroxyl radicals at the tips of the cruciforms. The data are consistent with the binding of a single antibody to an antigen-combining site. The results of this study compare favourably with the hydroxyl radical footprinting studies reported recently for a human cruciform binding protein (CBP), which binds at the base of the stem-loop structure and causes similar distortions of the stable cruciform structure. These studies indicate that the four-way junction of the cruciform possesses certain unique structural qualities that are antigenic; the association of this structural determinant with DNA replication and the existence of a novel cellular protein, CBP, of similar binding specificity as the antibody specificity support a role for cruciforms as important regulatory recognition signals in replication. PMID- 7473757 TI - The mutagenic processing of psoralen photolesions leaves a highly specific signature at an endogenous human locus. AB - To assess the role of a given genotoxic agent in the etiology of human cancers, it is useful to establish the mutational specificity of this agent. The aim of this study was to investigate whether the processing of psoralen photolesions, interstrand cross-links (CL) and monoadducts (MA), leaves a specific molecular signature in the mutational events produced at an endogenous locus, HPRT. Human lymphoblasts were treated by 4,5',8-trimethylpsoralen (Me3Pso) in association with a double irradiation protocol (365 plus 365 nm) which allows us to increase the proportion of CL for a given constant number of total photoadducts. The molecular spectrum of mutations at the HPRT locus induced in these conditions was compared to the previously reported spectra of mutations induced by the same psoralen in combination with a single irradiation of either 365 nm (induction of MA and a low proportion of CL) or 405 nm (producing almost exclusively MA). In all treatment conditions, base substitutions constitute the major type of Me3Pso photoinduced mutations. The majority of base substitutions involve a T residue preferably within a 5'-TpA sequence which corresponds to the favoured sites of psoralen photoadducts. In other words, the Me3Pso photolesions induce at the endogenous HPRT locus a high specific signature. Moreover, base substitutions have been essentially found in the non-transcribed strand of the HPRT gene suggesting that the psoralen photolesions are preferentially removed from the transcribed strand. In spite of the considerable difference between the proportion of lesions of both types (CL or MA) induced in different treatment conditions, the kind of mutations and their sequence distribution are similar suggesting that the mutagenic processing of psoralen CL and MA is similar at least for the steps resulting in base substitutions. PMID- 7473758 TI - A theory of DNA dissociation from the nucleosome. AB - Previous analysis of an elastic model of the nucleosome indicated that 10 bp end segments of DNA can exist in a continuum of mechanically stable trajectories ranging from complete winding on the histone octamer to complete unwinding. Stable states of 20 bp and 40 bp end segments, however, are grouped in bands separated by gaps where DNA trajectories are unstable. We extend these results to cover the entire range up to a complete nucleosomal turn, 80 bp. We find that 10 to 60 bp segments have states intermediate between fully wound and fully unwound that are mechanically stable. In striking contrast, there is no stable intermediate trajectory for 70 bp or 80 bp segments. Segments of these lengths constitute a two-state system. A 70 or 80 bp segment is either fully wound or fully unwound, and the population of these states is governed by Boltzmann's thermal distribution. We have found a plausible dissociation pathway from the fully wound to the fully unwound state for the 80 bp segment. In a ponderous breathing motion that breaks all contacts with the histone ponderous breathing motion that breaks all contacts with the histone surface, the segment climbs to an activation peak of about 12 kcal/mol, then rapidly straightens away from the histone core to complete dissociation. PMID- 7473759 TI - Requirement of a 3'-terminal stem-loop in in vitro transcription by an RNA dependent RNA polymerase. AB - Partially purified RNA-dependent RNA polymerase (RdRp) isolated from plants infected with turnip crinkle virus (TCV) is capable of template-dependent synthesis of TCV-associated RNAs. To determine the cis-sequences required for the synthesis of TCV satellite (sat-) RNA C (-) strands in vitro, templates containing interior deletions were subjected to transcription using RdRp-active fractions. Results indicated that the promoter for (-)-strand synthesis was contained within the 3'-terminal 29 bases of the (+)-strand. Structural probing by enzymatic digestion and chemical modification revealed the presence of a hairpin structure within this terminal region. Compensatory exchanges of four bases in the lower stem or alterations in the sequence and size of the loop region did not affect in vitro transcription, implying that the primary sequence in the loop and lower part of the stem is not important for interaction with the viral RdRp. However, single mutations in the base of the stem or double mutations in the upper stem strongly reduced template activity in vitro, suggesting that the stability of the hairpin is an important functional consideration. Relocation of the 3'-terminal 37 bases containing this stem-loop to inactive template RNA rendered the resultant hybrid RNA competent for in vitro transcription by RdRp activity, suggesting that the promoter for (-)-strand synthesis in vitro is completely contained within the 3'-terminal region. PMID- 7473760 TI - Contribution of hydrophobic residues to the stability of human lysozyme: calorimetric studies and X-ray structural analysis of the five isoleucine to valine mutants. AB - In order to understand the contribution of hydrophobic residues to the conformational stability of human lysozyme, five Ile mutants (Ile --> Val) in the interior of the protein were constructed. The thermodynamic parameters characterizing the denaturation of these mutant proteins were determined by scanning calorimetry, and the three-dimensional structure of each mutant protein was solved at high resolution by X-ray crystallography. The thermodynamic analyses at 64.9 degrees C and at pH 2.7 revealed the following. (1) The stabilities of all the mutant proteins were decreased as compared with that of the wild-type protein. (2) The changes in the calorimetric enthalpies were larger than those in the Gibbs energies, and were compensated by entropy changes. (3) The destabilization mechanism of the mutant proteins differs, depending on the location of the mutation sites. X-ray analyses showed that the overall structures of all the mutant human lysozymes examined were identical to that of the wild type protein, and only small structural rearrangements were observed locally around some of the mutation sites. The most striking change among the mutant proteins was found in the mutant protein, 159V, which contains a new water molecule in the cavity created by the mutation. The thermodynamic stabilities of the mutant proteins are discussed in light of the high-resolution X-ray structures of the wild-type and five mutant human lysozymes examined. PMID- 7473761 TI - Decomposition of interaction free energies in proteins and other complex systems. AB - A recent analysis of Mark and van Gunsteren has questioned the validity of separating different free energy components in proteins, or indeed in any complex system. The separability of free energy terms is re-examined from both a theoretical and a numerical perspective. Using a power series expansion of the free energy, it is found that the leading terms are free energy components that arise from individual contributions to the Hamiltonian ("in situ" free energies). The energetic part of an in situ free energy component is given by the ensemble average of the corresponding Hamiltonian component, while the leading term in the entropic part, which was missing in the analysis of Mark and van Gunsteren, is given by the mean square fluctuation. In addition there are correlations between fluctuations in each Hamiltonian component, which give rise to a coupling, or correlation entropy. A simple system, whose configurational degrees of freedom can be completely sampled, was examined in order to determine the relative sizes of these different contributions to the free energy. Under certain conditions, the change in system free energy observed when a particular component of the Hamiltonian is removed or altered is well approximated by the change in the in situ free energy of that component. In practical terms, this allows one in these cases to separate out different free energy contributions. PMID- 7473762 TI - Crystal structure of the SH2 domain from the adaptor protein SHC: a model for peptide binding based on X-ray and NMR data. AB - Src homology 2 domains (SH2) are protein molecules found within a wide variety of cytoplasmic signalling molecules that bind with high affinity to phosphotyrosyl (pY)-containing protein sequences. We report here for crystal structure of the SH2 domain from the adaptor protein SHC (Shc), which has been refined by restrained least-squares methods to an R-factor of 17.3% to 2.7 A. The overall Shc architecture is essentially similar to that determined in other SH2 domains but it shows significant differences in a number of loops, thus providing a molecular surface with no obvious secondary pocket. Based on the knowledge of the crystal structure of the protein a model for a low affinity Shc-bound peptide has been generated from nuclear magnetic resonance data in solution using transferred nuclear Overhauser enhancements as intramolecular distance restraints. The model shows that the tyrosine moiety binds Shc in a rather similar way to that observed for other SH2-peptide complexes, but that the residue in position +3 does not seem to make specific contact with the protein. An intermolecular crystallographic interaction occurs between the pY-binding site and the C terminal residues of a symmetry-related molecule. This crystal packing interaction suggests how inhibitory regulation could play a role in SHC activity. PMID- 7473763 TI - Strange bedfellows: interactions between acidic side-chains in proteins. AB - The oxygen atoms of two acidic side-chains are frequently found within hydrogen bonding distance of each other in proteins. Two distinct types of cases are common. In metal-binding sites, the oxygen atoms are brought near (average closest approach 3.0 A) by their common role as metal ligands. In a different location, either buried or on the protein surface, the two acidic groups can share a proton. The corresponding O-O distances in the latter case are shorter (usually 2.7 or less), and the geometry is typical of hydrogen-bonding interactions. The glucose/galactose-binding protein of Salmonella typhimurium provides an example of a well-ordered Asp-Glu pair on the surface of a protein with a very short O-O distance, at a pH of 7.0. Other instances have been found at pH values as high as 8.0, suggesting substantial alteration of the pKa involved. These observations have implications for the study of enzymes that use pairs of acidic residues in binding and catalysts. PMID- 7473764 TI - Ultrastructure and function of cellular components of the intercentral joint in the percoid vertebral column. AB - The intervertebral joint of the teleost, Perca flavescens, is formed by opposing amphicoelus centra whose rims are connected by external ligaments. The tissue, located within the space formed by these structures, is derived from the notochord and consists of the elastic externa, the fibrous sheath, and the notochordal cells. The cellular tissue within the joint has many characteristics of a stratified epithelium, and when examined with the transmission electron microscope, at least three morphologically distinct regions can be recognized. First, a peripheral single layer of columnar to squamous-shaped cells lies on a basement membrane immediately deep to the fibrous sheath. Second, several layers of cells, each containing a large central vacuole, occur. Third, in the deepest part of the joint, several layers of attenuated cells surround intracentral fluid filled lacunae and form a transverse septum across the joint. All cells in this tissue are interconnected by numerous desmosomes. Further, an extensive intermediate filament network exists in all three types of cells. The intermediate filament network in the vacuolated cells is arranged cortically around a membrane-bound vacuole, and suggests that these cells may act as passive cellular hydrostats. The squamous cells surrounding the joint lacunae are structurally similar to mammalian epidermal cells, and the intermediate filament network within them is layered parallel to the surface of the lacunae. The organization of these cells suggests that they are the tensile component of extracellular hydrostats within the intercentral joint. These cellular and extracellular hydrostats within the intercentral joint would function to resist the compressive and tensile stresses encountered during undulatory swimming. PMID- 7473765 TI - Incisal biting in the mountain beaver (Aplodontia rufa) and woodchuck (Marmota monax). AB - Analysis of synchronously recorded cine-radiographs and electromyograms in two rodents (Aplodontia rufa and Marmota monax) demonstrates that jaw movements and muscle activities during incisal functions are distinctly different from those found during mastication. Movements during incisal biting are primarily along the midline, accompanied by symmetrical activity of the jaw adductor muscles. Most biting cycles do not end in contact between upper and lower incisors. When contact does occur, the lower incisors are dragged along the lingual surfaces of the upper incisors. Cropping, or tip-to-tip occlusion of upper and lower incisors, was not observed. Sharpening of the lower incisors, a behavior which may be unique to the Rodentia, was recorded in both A. rufa and M. monax. During sharpening, the lingual surface of the lower incisor is dragged across the tip of the upper incisor producing a lingual wear facet. Like incisal biting, sharpening movements are primarily confined to the midline, although there may be lateral movements in some sharpening cycles. Sharpening cycles are among the most rapid cyclic movements recorded in mammals, as the mean frequencies of sharpening are 11 cycles/s in A. rufa and 8 cycles/s in M. monax. PMID- 7473766 TI - Cytological aspects of vascular invasion by the trophoblast of Calomys callosus in hepatic tissues. AB - Ectoplacental cones isolated from embryos on the seventh day of pregnancy were transplanted beneath the hepatic capsule of recipient adult animals to document the morphological patterns of vascular invasion by the trophoblast in the absence of the maternal environment and the influence of its peculiar vasculature. Females, and females and males of Calomys callosus, a cricetid rodent, were used, respectively, as embryo donors and recipient animals. Three to 5 days later, the grafted regions were excised and processed for light and electron microscopy. Invasion of the liver parenchyma by the trophoblast progressed along the vascular beds, associated with gradual phagocytosis of hepatic cells, greatly favoring the morphological characterization of invasive steps exhibited by the trophoblast to access the different kinds of vessels, to trespass the various vascular components and the different levels of the surrounding hepatic parenchyma. It is possible that either in utero during the establishment of embryomaternal circulation in early pregnancy or ex utero under experimental conditions, the trophoblast exhibits similar vascular invasion behavior. In view of this, our findings may contribute to a better understanding of trophoblast cell migration to the maternal blood supply as well as the role of the trophoblast in the establishment of the placental circulation during pregnancy. PMID- 7473767 TI - Distribution of cranial and rostral spinal nerves in tadpoles of the frog Discoglossus pictus (Discoglossidae). AB - We studied the peripheral nervous system of early tadpoles of the frog Discoglossus pictus using whole-mount immunohistochemistry. Double-labeling of muscles and nerves allowed us to determine the innervation of all cranial muscles supplied by the trigeminal, facial, glossopharyngeal, vagal, and hypoglossal nerves. The gross anatomical pattern of visceral, cutaneous, and lateral-line innervation was also assessed. Most muscles of the visceral arches are exclusively supplied by posttrematic rami of the corresponding branchiomeric nerves, the only exceptions being some ventral muscles (intermandibular, interhyoid, and subarcual rectus muscles). In the mandibular arch, the pattern of motor ramules of the trigeminal nerve prefigures in a condensed form the adult pattern, but the muscles of the hyoid arch are innervated by ramules of the facial nerve in a pattern that differs from that of postmetamorphic frogs. With respect to the nerves of the branchial arches, pretrematic visceral rami, typical of other gnathostomes, are absent in D. pictus. Instead, we find a separate series of posttrematic profundal visceral rami. Pharyngeal rami of all branchial nerves contribute to Jacobson's anastomosis. We provide a detailed description of the lateral-line innervation and describe a new ramus of the middle lateral-line nerve (ramus suprabranchialis). We confirm the presence of a first spinal nerve and its contribution to the hypoglossal nerve in D. pictus tadpoles. PMID- 7473768 TI - Progression of cardiac potassium current modification after brief exposure to reactive oxygen. AB - We reported previously that singlet oxygen (1O2), generated by illuminating the photosensitizer rose bengal (RB), suppressed the delayed rectifier potassium current (IK) in single frog atrial cells. Considering the brief lifetime of 1O2, one might expect IK modification to reach a steady-state soon after a brief exposure to RB-generated 1O2. Here we report that, contrary to expectations, tens of seconds can be required for IK to reach a new steady-state. We will use the term "progression" to refer to the component of current modification which occurs after cessation of illumination. To gain insight into the mechanism of progression, we investigated how its time course and magnitude were affected by (1) membrane potential during and following RB illumination, and (2) the level of IK activation during illumination. We found that conditions which favored the open state of the potassium channel also favored progression, increasing both its time course and magnitude. Illumination while IK was activated produced significant progression having a very slow time course (tens of seconds). By comparison, illumination when IK was not activated produced no progression: IK modification was completed during the 2 s illumination period. These findings suggest progression results from the kinetics of potassium channel state transitions rather than from a long-lived reactive intermediate produced during the initial 1O2 exposure. PMID- 7473769 TI - Asymmetrical electrically induced injury of rabbit ventricular myocytes. AB - Strong defibrillation-type electric field stimulation may injure myocytes when transmembrane potentials during the pulse exceed the threshold for membrane permeabilization. The location of injury may depend on intrinsic transmembrane potential or influx of calcium by "electro-osmosis" during the stimulation pulse in addition to the transmembrane potential changes induced by the pulse. We have studied injury by examining contracture and changes in transmembrane potential sensitive dye fluorescence induced by electric field stimulation (St) with a duration of 20 ms and strength of 16-400 V/cm in isolated rabbit ventricular myocytes. St of 100-150 V/cm produced injury in myocytes oriented parallel to the St field frequently without injuring myocytes oriented perpendicular to the field. Injury required calcium in the solution and was asymmetric, occurring first at the myocyte and facing the St anode in 100% of injured myocytes in normal Tyrode's solution. Injury depended significantly on whether the product of the electric field strength and myocyte length exceeded a threshold of 1.1 V (P < 0.05). Asymmetric injury at the end facing the anode was still present in 96% of injured myocytes for stimulation after depolarization by an action potential or 20 mM or 125 mM potassium, suggesting that intrinsic transmembrane potential is not responsible for asymmetry. In 125 mM potassium, eliminating calcium from the bathing solution during the St pulse and introducing calcium after the pulse decreased the fraction of injured myocytes in which injury occurred at the end facing the anode to 62%, suggesting that calcium influx by "electro-osmosis" at the myocyte end facing the anode contributes to asymmetry. Asymmetric injury at the end facing the anode was still present in 100% of injured myocytes after adding 1 mM tetraethylammonium chloride, indicating that asymmetry is not sensitive to the potassium channel blockade. For stimulation pulses stronger than 50 V/cm given after depolarization by an action potential, transmembrane potentials at both myocyte ends decayed after the initial deflection indicating that permeabilization occurred at both ends. In conclusion, injury depends on myocyte orientation and is asymmetric occurring first at the myocyte end facing the anode. Asymmetric injury is not explained by asymmetric permeabilization, is independent of the intrinsic transmembrane potential and may result from "electro osmosis" during the stimulation pulse. PMID- 7473770 TI - Ion specificity and stoichiometry of the cardiac inositol transporter. AB - Cardiac myocytes have a high affinity. Mg(2+)-dependent, electrogenic, Na inositol co-transporter on the plasma or sarcolemma (SL) membrane (Rubin and Hale, 1993). The ion specificity and stoichiometry of this transport process is unknown. Using bovine cardiac SL vesicles, we have determined that transmembrane movement of myo-[3H]inositol requires Na+ and is not supported by K+ or Li+. Furthermore, replacing Cl- with a non-permeant anion has no effect on inositol transport. Carrier-mediated inositol efflux indicated that efflux was Na(+) dependent and electrogenic but independent of extravesicular Mg2+ in the efflux media. The transport stoichiometry of 1 Na+ for one inositol was derived by determining the null point of myo-[3H]inositol flux under conditions where the inside and outside concentration (ratio) of myoinositol and Na+ were controlled. The results suggest that Na+, inositol, and Mg2+ bind to the same side of the membrane for transport to occur and that the stoichiometry of Na+ and inositol transport is 1:1. PMID- 7473771 TI - Messenger RNA for the guanine nucleotide-binding regulatory protein (G protein) is reduced in the acute ischemic myocardium. AB - It has been reported that the function of the guanine-binding regulatory protein (G protein), especially the alpha subunit of the stimulatory G protein (Gs alpha), in myocardium is decreased with acute ischemia. However, it is unclear whether this decrease is due to transcriptional or post-transcriptional changes. Moreover, no studies have examined the distribution of G protein mRNA in ischemic myocardium using in situ hybridization. The purpose of this study was to explore alterations in mRNA of G proteins (Gs and Gi) in ischemic hearts using in situ hybridization. We measured the levels of mRNA for Gs alpha and Gi alpha in ischemic and non-ischemic myocardium by in situ hybridization using a radioisotope imaging system. We compare these mRNA levels in ischemic and non ischemic myocardium with Northern blot analysis and the protein levels of G proteins by Western blot analysis. The mRNA for Gs alpha and Gi alpha was distributed diffusely in normal hearts. Levels of mRNA detected by in situ hybridization were substantially reduced by acute ischemia, and these results were confirmed by Northern and Western blot analysis. These results suggest that decreased levels of mRNA and protein for G proteins may underlie the impaired function of the receptor--G protein--adenylate cyclase system in ischemic myocardium. In addition, quantitative evaluation of mRNA is possible by in situ hybridization and correlates well with Northern analysis. PMID- 7473772 TI - Decreased contractile responses to isoproterenol in isolated cardiac myocytes from aging guinea-pigs. AB - We have characterized the age-related changes of contractility and beta adrenoceptor function in isolated cardiac myocytes from guinea-pigs. We used either adult animals from 2 to 14 weeks of age, where body weight increases linearly with age, or senescent ones aged between 53-65 weeks. There was some indication of a decrease in contractility in maximum Ca2+ with age, with significant differences between a young (< or = 4 weeks, weight < 400 g) and aged (> or = 8 weeks, weight > 600 g) group in contraction amplitude expressed as percentage shortening (but not when expressed as micron change in length) or contraction and relaxation velocities. This decline was continued into senescence, and ANOVA showed a significant difference between the three groups for contraction amplitude (percentage shortening, 12.2 +/- 0.9%, young, n = 31; 9.5 +/- 0.6%, n = 28 aged; 6.7 +/- 0.8%, n = 6, senescent; P = 0.005), and contraction or relaxation velocities (P < 0.001). There was a more pronounced decline in maximum response to isoproterenol with age. ANOVA for the maximum isoproterenol response for the three divisions showed significant differences for percentage shortening (11.8 +/- 0.7%, n = 30, young; 7.9 +/- 0.5%, n-28, aged and 5.5 +/- 1.1%, n = 6, senescent; P < 0.001), velocities of contraction (P < 0.001) and relaxation (P < 0.001), and normalized velocities of contraction (P < 0.001) and relaxation (P < 0.01) at maximum isoproterenol, as well as in ISO EC50 (P < 0.001) and isoproterenol/Ca2+ ratio (P < 0.02). A general decrease in contractility of the myocyte occurs as the animal ages, with maximum contraction amplitude being reduced and velocity of contraction and relaxation slowed. The effect was more pronounced for beta-adrenoceptor stimulation than for high Ca2+, suggesting a specific lesion in the adenylate cyclase related pathway. Much of the change occurred between the young adult (< or = 4 weeks) and the aged adult (> or = 8 weeks), although the trend was continued in senescent animals (> 52 weeks). PMID- 7473773 TI - Angiotensin II is a potent stimulator of MAP-kinase activity in neonatal rat cardiac fibroblasts. AB - We have previously shown that angiotensin II (AII) is a mitogen for neonatal rat cardiac fibroblasts. However, the signaling events that lead to fibroblast cell growth in response to AII remain to be elucidated. Mitogen-activated protein (MAP) kinases are cytosolic serine/threonine kinases which have been shown to be activated in quiescent cells by diverse growth stimuli, thereby being linked to growth regulatory pathways. This study was designed to determine whether MAP kinase activation occurred in response to AII/receptor coupling in neonatal rat cardiac fibroblasts and the role of MAP-kinase activation in the AII-induced proliferation of these cells. Immunoblot analysis of MAP-kinase isoforms revealed predominantly p44 with less p42 MAP-kinase in rat cardiac fibroblasts. Both isoforms were activated upon stimulation of the cells with AII for 5 min or platelet derived growth factor-BB for 10 min. Angiotensin II stimulated MAP kinase in a dose-dependent fashion with an EC50 of 2.5 nM. Two minutes following stimulation with 1 microM AII MAP-kinase activity increased from 90 +/- 17.9 to 477.5 +/- 75.9 pmol/min/mg protein, P < 0.05, n = 4. A smaller, sustained, secondary increase in MAP-kinase activity from 37.7 +/- 5.3 to 110.9 +/- 15.3 pmol/min/mg protein, P < 0.05, n = 4, was observed in response to AII between 120 150 minutes following receptor occupancy. The responses to AII were markedly attenuated by the AT1 receptor antagonist EXP3174. Stimulation of the cells with carbachol induced the first but not the second phase of MAP-kinase activity and this compound had no effect on cellular growth. The second phase of MAP-kinase activity 2-2.5 h after AII stimulation, paralleled data demonstrating that a 2-3 h receptor occupancy with AII was necessary to induce DNA synthesis and fibroblast proliferation. These results indicate that AII stimulates a biphasic activation of MAP-kinase by the AT1 receptor and that this pathway may participate in the AII induced mitogenic response in cardiac fibroblasts. PMID- 7473774 TI - Two different mechanisms of noradrenaline release during normoxia and simulated ischemia in human cardiac tissue. AB - Species-related differences in the mechanisms of noradrenaline release during normoxia and myocardial ischemia emphasize the need for studies on human hearts. Therefore, the mechanisms of noradrenaline release were investigated during normoxia and energy depletion in incubated human atrial tissue and compared to the release characteristics in normoxic and ischemic rat heart. Potential differences of atrial versus ventricular myocardium were assessed by comparing catecholamine release during electrical stimulation and ischemia in isolated rat atrium with release characteristics in the intact perfused heart. The overflow of endogenous noradrenaline and its deaminated metabolite dihydroxyphenylethyleneglycol (DOPEG) were determined by high pressure liquid chromatography and electrochemical detection. During normoxia noradrenaline release was evoked by electrical field stimulation. Stimulation-induced noradrenaline release depended on the extracellular calcium concentration in both species and was almost completely suppressed under calcium-free conditions. The release was significantly inhibited by neuronal (N-type) calcium channel blockers such as omega-conotoxin (100 nmol/l) and cadmium chloride (100 mumol/l), indicating a predominant role of N-type calcium channels in exocytotic noradrenaline release from sympathetic neurons in human and rat heart. Desipramine (100 nmol/l) enhanced the overflow of noradrenaline evoked by electrical stimulation in both species by blocking neuronal catecholamine uptake (uptake1). Myocardial ischemia was caused by interruption of perfusion flow in rat heart and simulated by anoxic and glucose-free incubation in human and rat atrial tissue. Ischemia- and anoxia-induced noradrenaline release in rat heart and human atrial tissue was unaffected by varying extracellular calcium concentrations and occurred even after omission of calcium and addition of EGTA (1 mmol/l). In both species neither omega-conotoxin (100 nmol/l) nor cadmium chloride (100 mumol/l) affected ischemia-induced noradrenaline overflow in both rat heart and atrium as well as in human atrium. In human and rat atrial tissue, blockade of energy metabolism in the presence of oxygen (cyanide model) resulted in a desipramine-sensitive release of noradrenaline, which was accompanied by DOPEG overflow, indicating increased axoplasmic noradrenaline concentration. The data imply a dual mechanism of noradrenaline release in the human heart. During normoxia noradrenaline release is modulated by neuronal calcium influx indicating exocytotic release. Ischemia-induced noradrenaline release, however, is independent of calcium and inhibited by uptake1 blockade suggesting nonexocytotic release mechanism. The characteristics of noradrenaline release in human atrial tissue provide evidence for carrier-mediated release of noradrenaline from sympathetic neurons operative in the ischemic human myocardium. PMID- 7473775 TI - Isolation of rat heart endothelial cells and pericytes: evaluation of their role in the formation of extracellular matrix components. AB - In order to facilitate investigation of the cells responsible for overproduction of type VI collagen in the extracellular matrix surrounding the capillaries of diabetic rat myocardium, procedures have been developed for the isolation from this tissue of endothelial cells as well as a cell type identified as pericytes. This was accomplished by enzymatic and mechanical disruption of ventricles from young rats (125 g) followed by removal of myocytes through their nonadherence to tissue culture surfaces. Endothelial cells were separated by fluorescence activated cell sorting after staining with rhodamine-labeled acetylated low density lipoprotein and were identified by their monolayer growth pattern, reaction with anti-von Willebrand factor and the ability to form capillary-like tubes induced by low serum concentration. Pericytes were purified by selective scraping for removal of other cell types and were identified by their irregular shape, overlapping growth pattern at confluence, reaction with anti-smooth muscle actin and content of GLUT4 glucose transporter. Fibroblasts, visualized after staining with rhodamine-labeled alpha 2-macroglobulin, were only rarely detected. Analysis of collagen by immunoblotting indicated formation by both cell types of alpha 1(IV) collagen as well as the three subunits of type VI (alpha 3 at 205 kDa and alpha 1 plus alpha 2 at 150 kDa). Both endothelial cells and pericytes demonstrated transcripts for types VI, IV and I collagen, as well as fibronectin, but while the level of the mRNA for type IV collagen was higher in pericytes than in endothelial cells, the reverse was true for collagens VI and I and fibronectin. These observations suggest that both endothelial cells and pericytes contribute to formation of the myocardial capillary matrix, but that changes involving only type VI collagen, such as occur in diabetic cardiomyopathy, may reflect a response primarily of endothelial cells. PMID- 7473777 TI - Mechanisms of the circadian regulation of beta-adrenoceptor density and adenylyl cyclase activity in cardiac tissue from normotensive and spontaneously hypertensive rats. AB - Circadian variation in beta-adrenoceptor density and adenylyl cyclase activity was studied in myocardium from normotensive Wistar-Kyoto rats (WKY) and spontaneously hypertensive rats (SHR). In SHR cardiac beta-adrenoceptor density was significantly lower than in WKY. This reduction in total beta-adrenoceptors was exclusively due to a loss in the beta 1-subtype. Total and beta 1 adrenoceptor density in ventricles from both strains exhibited significant circadian variation with peaks occurring in the middle of the light and dark periods, whereas the beta 2-subtype did not show rhythmicity. Similarly, stimulation of adenylyl cyclase via total beta-adrenoceptors and via the beta 1 subtype was circadian time dependent, but circadian peaks occurring at the beginning of the light and dark periods were 6 h apart from those in beta adrenoceptor density. Rhythmicity in the formation of cAMP was observed under basal conditions and after stimulation by isoprenaline or a forskolin derivative, whereas addition of manganese-ions abolished the circadian variation. In conclusion, the present study demonstrates that in ventricular tissue from WKY and SHR circadian rhythms observed in total beta-adrenoceptor density are due exclusively to variations in the beta 1-subtype. Since peaks in cAMP formation coincided with troughs in beta-adrenoceptor number, cAMP mediated phosphorylation of beta-adrenoceptors enhancing their down-regulation, could be involved in the circadian regulation of myocardial beta-adrenoceptor density. Rhythmicity in cAMP formation itself seems to involve coupling of G-proteins, because manganese ions abolished the circadian variation. PMID- 7473776 TI - Cardiac Na+/K+ ATPase activity and its relation to myocardial glutathione status: studies in the rat. AB - Cardiac Na+/K+ ATPase (which has previously been suggested to be regulated by a thiol-dependent process) is inactivated during ischemia. Moreover, myocardial glutathione (which controls the redox state of protein-thiol groups) is depleted during ischemia and thus may contribute to the changes in Na+/K+ ATPase activity. The objectives of the present study were to use the rat to: (1) pharmacologically manipulate myocardial glutathione to mimic ischemia-reperfusion-induced glutathione depletion and (2) determine (in the absence of ischemia) the relationship between myocardial glutathione content and Na+/K+ ATPase activity. Tissue glutathione was depleted by injecting rats with diethylmaleate (0, 55, 110, 215, 430 or 860 mg/kg; i.p.) 30 min before study. Total glutathione content fell from 1.72 +/- 0.03 to 1.66 +/- 0.04, 1.50 +/- 0.05, 0.93 +/- 0.03, 0.21 +/- 0.02 mumol/g wet weight, respectively. There was a linear correlation (r = 0.96) between Na+/K+ ATPase activity and glutathione content in diethylmaleate-treated animals. A separate group of animals were treated with phorone (0, 25, 50, 100, 150, 200 or 250 mg/kg; i.p. 120 min before study) which also depletes glutathione. Myocardial glutathione fell from its control value of 1.74 +/- 0.03 to 1.52 +/- 0.04, 1.35 +/- 0.06, 1.14 +/- 0.05, 0.84 +/- 0.04, 0.64 +/- 0.04, 0.54 +/- 0.03 mumoles/g wet weight, respectively. In these animals Na+/K+ ATPase activity was also linearly-related (r = 0.98) to glutathione content. We also characterized the temporal relationship between diethylmaleate-induced glutathione depletion and Na+/K+ ATPase activity. Diethylmaleate (216 mg/kg; i.p.) was given to rats and at various times (0-200 min), after administration the hearts were removed and assayed for glutathione. During the first 30 min after administration glutathione fell from 1.74 +/- 0.03 to its lowest value of 0.90 +/- 0.04 mumol/g wet weight. It then progressively recovered to within control levels by 150 min after administration. Na+/K+ ATPase activity paralleled the recovery of glutathione status. In additional studies, the cell-permeant glutathione analogue YM 737 (glutathione isopropyl ester: 1 mmol/kg, i.p.) accelerated the recovery of tissue glutathione and evoked a commensurate and parallel increase in the rate of recovery of Na+/K+ ATPase activity. Our studies suggest there is a close coupling between tissue glutathione content and Na+/K+ ATPase activity and that this may be an important factor in ischemia and reperfusion-induced changes in Na+/K+ ATPase activity. PMID- 7473778 TI - Molecular changes of titin in left ventricular dysfunction as a result of chronic hibernation. AB - Cardiomyocytes of chronic hibernating myocardium are affected by partial to complete loss of sarcomeres, accumulation of glycogen, adaptations in size and shape of mitochondria, reorganisation of nuclear chromatin and depletion of sarcoplasmic reticulum. The nature of these changes, which from a purely morphologic viewpoint are akin to dedifferentiation, needed further clarification at the molecular level. For this purpose we have studied the expression and reorganization of titin, one of the earliest markers of cardiomyocytes differentiation. By use of monoclonal antibodies, recognizing different epitopes distributed over the whole length of the titin molecule, we were able to detect changes in its molecular organization as a result of chronic hibernation. The epitopes of the titin molecule attached to the Z-disc and those present close to the M-line remained detectable at all stages of hibernation, while epitopes at the A-I junction and parts of the myosin anchoring region of the molecule became masked or were lost. A fragmented or punctuated appearance of the titin staining pattern with antibodies to A-I junction related epitopes is found in cells which we consider to represent a more advanced stage of dedifferentiation. Changes in the distribution of the titin molecule or its molecular environment in hibernating myocardium resemble at least in part changes occurring during muscle cell differentiation, although in reversed order. PMID- 7473779 TI - Myocardial metabolism and function in acutely ischemic and hypoxemic isolated rat hearts. AB - We tested the hypothesis that residual oxygen supply during acute low-flow ischaemia or hypoxemia is a major regulator of myocardial performance, metabolism and recovery. Rat hearts were exposed for 20 min to either ischemia (coronary flow reduced to 10% of baseline), hypoxemia (oxygen content reduced to 10% baseline) or a "mixed" condition (combined ischaemia and hypoxemia). The oxygen supply (coronary flow x oxygen content) was matched in all groups (n = 16 per group). Hypoxemic hearts had the highest performance (systolic and developed pressures, +/- dP/dtmax and oxygen uptake) and content of IMP and AMP. Ischaemic hearts had the highest content of ATP, phosphocreatine, adenine nucleotides and purines. As flow and/or oxygenation were restored, post-ischemic hearts showed better functional and metabolic recovery than post-hypoxemic ones. "Mixed" hearts were more similar to hypoxemic ones during oxygen shortage but to ischemic ones during recovery. We conclude that as oxygenation is critically limiting, coronary flow is relatively more important than oxygen supply in determining myocardial function, metabolism and recovery, most likely secondary to changes in the metabolism of diffusible substances. PMID- 7473780 TI - Dioctanoyl-glycerol inhibits L-type calcium current in embryonic chick cardiomyocytes independent of protein kinase C activation. AB - Diacylglycerol analogs and phorbol esters are used as protein kinase C (PKC) activators to investigate the effect of PKC on L-type calcium current [ICa(L)] in cardiomyocytes. 1-2-dioctanoyl-sn-glycerol (diC8) is a potent analog of diacylglycerol (DAG) which produces positive inotropic effects in guinea-pig atria cardiomyocytes via PKC activation. DiC8 effect on ICa(L), recorded (at 25 degrees C) in whole-cell voltage-clamp, was measured in 17-day-old embryonic chick cardiomyocytes in culture. ICa(L) was recorded in Na+, K(+)-free solution (external) and Ca(2+)-, K(+)-free solution (pipette), with depolarizing steps (to +10 mV) applied from a holding potential of -40 mV. Perfusion with different concentrations of diC8 (from 0.1 to 100 microM) inhibited ICa(L) in a dose dependent manner, with half-maximal inhibition occurring at 12.5 microM. The effect of diC8 occurred rapidly, the effect beginning within 2 min and being completed within 5 min. In order to determine if the inhibitory effect of diC8 on ICa(L) was through activation of PKC, 25 microM diC8 was applied after pre incubation of the cardiomyocytes with the PK inhibitors staurosporine (1 microM) or H-7 (50 microM). The effect of diC8 was not prevented by staurosporine or H-7. To further rule out the involvement of PKC in the action of diC8, experiments were performed using another analog of DAG, 1-oleyl-2-acetyl-glycerol (OAG, 50 microM) and Angiotensin-II (A-II, 0.1 microM). OAG failed to produce any effect on ICa(L). A-II, believed to act by activation of PKC did not affect ICa(L) within a test period of 8 min.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7473781 TI - The effects of compensated cardiac hypertrophy on dihydropyridine and ryanodine receptors in rat, ferret and guinea-pig hearts. AB - The number of dihydropyridine and ryanodine receptors (DHP-R and RyR) has been measured in control and hypertrophied ventricles from rats, guinea pigs and ferrets to determine whether these two channels contribute to the alterations in excitation-contraction coupling (ECC), and in Ca2+ transient during compensated cardiac hypertrophy. We found that ventricular hypertrophy did not change the density of DHP-R. Mild hypertrophy did not alter the density of RyR in the rat but decreased it in the guinea-pig and in the ferret (30% and 36%, respectively). Severe hypertrophy decreased the density of RyR by 20% in the rat and by 34% in the guinea-pig. Therefore, the decrease is greater in ferret and guinea-pig hearts than in rat heart. We conclude that the sarcoplasmic reticulum (SR) Ca2+ release channels but not the L-type Ca2+ channels could contribute to the slowing of intracellular Ca2+ movements and to the reduced velocity of shortening of the hypertrophied hearts. We suggest that, in the guinea pig and ferret hearts which express only the beta myosin heavy chain (MHC) isoform, the reduced velocity of shortening during hypertrophy is related to the decrease in RyR density, whereas in the rat, it is regulated primarily via a shift in the MHC isoform, except in severe hypertrophy in which the moderate decrease in RyR would also be involved. PMID- 7473782 TI - The effect of insulin and FFA on myocardial glucose uptake. AB - In addition to direct stimulation of glucose uptake and metabolism in cardiac myocytes, insulin inhibits lipolysis and, thereby, reduces serum free fatty acid (FFA) concentrations. This, in turn, has been suggested to enhance myocardial glucose utilization. To study the mechanism of insulin action on myocardial glucose uptake (MGU) in vivo, five patients with stable coronary artery disease were studied with positron emission tomography (PET) and [18F]FDG. All patients underwent two PET studies after a 12-h fast, once during low serum FFA but high insulin concentrations (during insulin clamp), and once during low serum FFA and low insulin concentrations (in the fasting state after two oral doses of 250 mg of an antilipolytic drug, acipimox). The MGU in the normal myocardium was measured using dynamic PET imaging. Plasma glucose concentrations were comparable during the insulin clamp and after administration of acipimox (5.0 +/- 0.4 v 5.2 +/- 0.3 mmol/l, n.s.). Serum insulin concentrations were high during clamp but remained in low fasting concentrations after acipimox (74 +/- 9 mU/l v 6 +/- 5 mU/l, P = 0.0001). Serum FFA concentrations were similar during both approaches (230 +/- 110 v 200 +/- 40 mumol/l, respectively, n.s.). No difference in cardiac work load was detected between the approaches. The calculated MGU values in normal myocardium were similar during both approaches (57 +/- 23 mumol/min/100 g v 61 +/- 14 mumol/min/100 g, respectively, n.s.). The MGU values correlated inversely to serum FFA concentration (r = -0.87, P = 0.001) and directly to myocardial work load (r = 0.73, P = 0.016) but not to serum insulin concentration.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7473783 TI - Is lactate-induced myocardial ischaemic injury mediated by decreased pH or increased intracellular lactate? AB - The detrimental effect of exogenous lactate during ischaemia on post-ischaemic contractile function may be mediated either by a lactate-induced intracellular H+ load or by an increase in intracellular lactate. To distinguish between these two mechanisms, isolated rat hearts were perfused with lactate or pyruvate during low flow ischaemia, the rationale being that both would decrease H+ efflux via lactate/H+ cotransport and lead to decreased pH, but only exogenous lactate would decrease lactate efflux and lead to increased intracellular lactate. 31P NMR spectra were acquired sequentially while hearts were subjected to 32 min low flow (0.5 ml/min) ischaemia and 32 min reperfusion. During ischaemia, hearts were perfused with Krebs-Henseleit buffer containing 11 mM glucose (controls) or 11 mM glucose plus either 10 mM lactate or 10 mM pyruvate. Reperfusion of all hearts was with buffer containing only glucose. Intracellular volume, estimated to be 0.52 ml/heart using 31P NMR spectroscopy with phosphonate space markers, did not change under any of the ischaemic conditions during the protocol. Control and pyruvate hearts recovered approximately 85% of pre-ischaemic contractile function, but there was no recovery of function in lactate hearts. This lack of recovery correlated with a 57% loss of ATP during ischaemia, which was significantly greater (P < 0.001) than the 41% loss of ATP in control and pyruvate-perfused hearts. End-ischaemic intracellular pH was 6.60 in both lactate perfused and control hearts, but significantly lower (P < 0.05) at pH 6.43 in pyruvate-perfused hearts. Both exogenous pyruvate and lactate should have decreased H+ efflux, however the higher pH in the lactate-perfused hearts could be explained by a 60% inhibition of glycolysis, determined by measurement of myocardial lactate production. Thus, the intracellular pH during ischaemia does not necessarily predict the extent of myocardial injury. We propose that lactate induced damage is a consequence of increased intracellular lactate leading to inhibition of glycolysis, presumably via an increased NADH/NAD ratio. This study highlights the important role of glycolysis in the ischaemic rat heart. PMID- 7473784 TI - The unique amino-terminal peptide of cardiac troponin I regulates myofibrillar activity only when it is phosphorylated. AB - Protein kinase A (PKA) dependent phosphorylation of C-protein and cardiac troponin I (cTnI) is known to be associated with a reduced sensitivity to Ca2+. We have investigated the relative importance of each of these sites of phosphorylation in this effect by use of extraction/reconstitution experiments and mutagenesis of recombinant cTnI. Conditions developed for extraction of troponin (Tn) complex also resulted in extraction of C-protein. A truncated cTnI (cTnI/NH2) lacking the 32 amino acids in the unique amino terminal extension of cTnI was engineered and expressed. In contrast to native cTnI, cTnI/NH2, which lacks Ser23 and Ser24, was not phosphorylated by PKA either in pure form or after incorporation into the myofilament lattice. The relation between pCa (-log molar free Ca2+ concentration) and MgATPase activity of non-phosphorylated native myofibrils or non-phosphorylated myofibrils reconstituted with cTnI, but lacking C-protein, was the same and could not be distinguished from that of control or PKA-treated myofibrils into which we exchanged cTnI with cTnI/NH2. However, PKA dependent phosphorylation of either native myofibrils or reconstituted myofibrils containing cTnI but lacking C-protein resulted in an identical and significant rightward shift of pCa50 (half-maximally activating pCa) in the pCa-activity relationship. Our results strongly indicate that phosphorylation of cTnI at Ser residues in the unique amino terminal extension of the molecule is both necessary and sufficient for the decrease in myofilament Ca(2+)-sensitivity associated with PKA-dependent phosphorylation. PMID- 7473785 TI - Proliferating cell nuclear antigen (PCNA) detection of cellular proliferation in hypothyroid and hyperthyroid rat hearts. AB - Immunocytochemical techniques examining the expression of cell proliferation related markers such as proliferating cell nuclear antigen (PCNA), may be employed to provide visual and quantitative evidence of cell proliferation. The efficacy of this method in frozen samples was tested on sections of 2-day-old rat heart. Mouse monoclonal anti-PCNA and goat anti-mouse IgG2a peroxidase-conjugated antibodies were applied to tissue cross-sections. PCNA positive (PCNA+) cells were evident in these sections as intensely stained, dark brown nuclei. This method was then applied to the hearts of four groups of adult rats (1) 1 month hyperthyroid, (2) 1 month hypothyroid, (3) 6 days hyperthyroid following 1 month hypothyroidism, or (4) euthyroid, in order to assess the ability of identifying and quantifying PCNA+cell types (myocyte, endothelial, or remaining, "other" cells). Serial cross-sections were prepared, half of the sections processed using the PCNA technique, while corresponding sections were stained with hematoxylin and eosin. Identification of PCNA+cell types was made by comparison of PCNA with H&E sections. PCNA labeling indices for a given cell type were expressed as the proportion of PCNA+nuclei per 1000 nuclei. No PCNA+myocyte nuclei were observed in any of the adult hearts. Only hypothyroid/hyperthyroid demonstrated significantly higher PCNA labeling of endothelial and "other" nuclei compared to euthyroid. This finding agrees with previous morphometric data which suggested capillary growth in these hearts. Results demonstrate that quantitation of PCNA expression on frozen heart tissue is possible, and can be used to provide direct, discernible evidence for cell proliferation. PMID- 7473787 TI - Rapid conversion from beta-MHC to alpha-MHC mRNA expression in embryonic rat ventricle cultured in oculo is not dependent on thyroid hormone or testosterone. AB - We investigated the expression of myosin heavy chain (MHC) isoenzymes in embryonic rat ventricles cultured in the anterior eye chamber of an adult rat. In oculo, these grafts beat and mature in an environment where the hormonal milieu can be manipulated. S1 nuclease protection assays were performed on pooled samples of ventricle grafts and compared to normally growing ventricles. At the time of grafting (embryonic day 12, E-12), 23 +/- 4% of the MHC mRNA was of the alpha isoform. While the proportion of ventricular alpha-MHC mRNA did not increase in utero, embryonic ventricles cultured in oculo showed a rapid increase in the relative amount of alpha-MHC mRNA expression (to 84 +/- 10% by 3 days and 86 +/- 5% by 8 days in oculo). alpha-MHC mRNA expression predominated through 8 weeks of culture in oculo, being 76% at 8 weeks in oculo. Additional experiments were performed to determine whether the rapid conversion to alpha-MHC expression resulted from exposure to adult levels of testosterone or thyroid hormone. Reduction of testosterone exposure to nondetectable levels by host orchiectomy did not affect the rapid conversion to alpha-MHC mRNA expression. Exposure to a hypothyroid milieu (i.e., PTU-treated hosts) decreased but did not prevent the conversion from beta- to alpha-MHC mRNA expression at 8 days in oculo; with 83% of the MHC mRNA being of the alpha isoform in hypothyroid hosts compared to 95% in euthyroid hosts. After 8 weeks of culture in hypothyroid hosts, however, alpha MHC mRNA expression was undetectable in grafted ventricles. These data suggest that E-12 myocardial grafts respond to the hormonal milieu of an adult rat with rapid conversion from beta- to alpha-MHC mRNA expression and that alpha-MHC expression in early developing heart may show reduced sensitivity to downward modulation by a hypothyroid hormonal milieu. PMID- 7473786 TI - Direct detrimental effects of L-arginine upon ischemia--reperfusion injury to myocardium. AB - The effects of L-arginine on recovery of myocardial contractile function and oxidative metabolism were investigated in a model of reversible global normothermic, ischemic injury using an isolated, buffer-perfused rabbit heart preparation. One mM L-arginine was infused into hearts for 2 min at the onset (group 1) of a 35 min period of ischemia or at the onset of reperfusion (group 2). In non-ischemic hearts, L-arginine caused a slight increase in developed pressure but had no effects on diastolic pressure, oxygen consumption (MVO2), coronary flow, or lactate production. When administered either before or after ischemia-reperfusion. L-arginine caused a significant increase in the diastolic pressure-volume relationship (PVR) and decline in systolic function when compared to untreated control hearts receiving the same ischemic injury. Recovery of MVO2 and high energy phosphates (phosphocreatine and ATP), measured by 31P-NMR spectroscopy, were significantly impaired in L-arginine-treated hearts compared to reperfused control hearts. Lactate release on reperfusion was also higher in both arginine-treated groups. Nitric oxide release into the coronary circulation (measured in separate experiments by the conversion of [15N]L-arginine to [15N]nitrate/nitrite using gas chromatography/mass spectroscopy) was not increased by L-arginine administration. Thus, we conclude that L-arginine acts synergistically with ischemia reperfusion to augment myocardial injury, which includes inhibition of oxidative metabolism and mitochondrial function. PMID- 7473788 TI - Gene expression in a swine model of right ventricular hypertrophy: intercellular adhesion molecule, vascular endothelial growth factor and plasminogen activators are upregulated during pressure overload. AB - We have investigated the molecular changes which occur during pressure overload hypertrophy of the RV in swine. Animals were banded on the pulmonary artery so that right ventricular pressure was increased two-fold. The heart was harvested at 3, 7, 24 and 72 h after surgery. Between 7 and 72 h there was evidence of muscle damage and inflammation. Northern blot experiments showed that pressure overload induced a transient increase in the expression of the immediate early genes and in the developmentally regulated atrial natriuretic factor and skeletal muscle alpha actin genes. Consistent with the histological observations of inflammation, increases in the expression of the gene for intercellular adhesion molecule, which encodes a protein involved in the binding of leukocytes by endothelial cells and myocytes, was observed between 3 and 24 h. In addition, the expression of vascular endothelial growth factor, a growth and permeability factor specific for endothelial cells was increased at 3 and 7 h of pressure overload. An increase in the expression of urokinase plasminogen activator and its inhibitors, plasminogen activator inhibitors I and II, was also observed between 3 and 24 h. This was associated with an increase in urokinase activity in the myocardial tissue. These results indicate that hypertrophy in a large mammal such as swine induces a program of gene expression similar to that previously described in rodents and suggests that up-regulation of a variety of other genes is an early response to pressure overload. PMID- 7473789 TI - Differential influence of extracellular and intracellular pH on K+ accumulation in ischaemic mammalian cardiac tissue. AB - The separate and the combined influence of lowering extracellular and intracellular pH on the extracellular accumulation of K+ ions in ischaemic mammalian cardiac tissue was investigated. Isolated guinea-pig papillary muscles were superfused in vitro while micro-electrode measurements of the transmembrane potentials and of the extracellular pH and K+ activity and of the intracellular pH and Na+ activity were performed. Muscles were reversibly subjected to spaced episodes of simulated ischaemia, and ischaemic K+ accumulation was measured. During normal superfusion, acidification of the intracellular pH (pHi approximately 6.8) effected by transient exposure to NH+4 containing superfusate was associated with an amiloride-sensitive rise of the intracellular Na+ activity (aiNa) from 5.1 +/- 0.5 to 19.5 +/- 2.4 mM. This increase was associated with a transient hyperpolarization of the membrane potential and shortening of the action potential duration from 190 +/- 7 to 111 +/- 16 ms. When a similar intracellular acidosis was induced 3 to 5 min before imposing ischaemia, extracellular K+ accumulation was increased compared to ischaemia alone. Surface K+ activity (asK) measured after 10 min of ischaemia was 9.6 +/- 0.8 mM following intracellular acidification v 6.4 +/- 0.4 mM following control superfusion. When the extracellular pH (pHo) was decreased to 6.85 prior to the ischaemic insult, extracellular K+ accumulation was not different from that observed during ischaemia after control superfusion (5.1 +/- 0.5 v 4.7 +/- 0.3 mM, respectively, after 10 min of ischaemia). When combined intracellular and extracellular acidosis was produced before ischaemia, the enhancing influence of lowered pHi on K+ accumulation was not observed. In these acid loading conditions, aiNa rose to the same level as observed following NH+4 withdrawal without simultaneous external acidification (26.9 +/- 2.0 mM v 23.9 +/- 2.0 mM, respectively). The results indicate that decreasing pHi before ischaemia accelerates the ischaemic increase of extracellular K+. On the other hand, although prior extracellular acidification by itself does not directly influence ischaemic K+ accumulation, it is in some way protective by reducing the intracellular proton-stimulated K+ efflux. This reduction seems not to be due to decreased cellular Na+ loading. PMID- 7473790 TI - The future of our water supply. PMID- 7473791 TI - The decline of Braille. PMID- 7473792 TI - Helping the person who is visually impaired: concerns, questions, remedies, and resources. AB - Information that you need to know to help meet the needs of individuals who are blind or visually impaired can be obtained by contacting appropriate local private or public agencies. A comprehensive analysis of losses associated with visual deterioration provides the foundation to restore deficits. A holistic approach to helping restore the person with visual impairment to independence includes skill development, sensory training, emotional strengthening, and sighted-world integrating. PMID- 7473793 TI - Changing practice patterns in general ophthalmology. AB - 1. The orientation of general ophthalmic practice in the United States has undergone a dramatic metamorphosis from frequent use of hospital inpatient services, to primarily an outpatient subspecialty. 2. Improved surgical techniques and instrumentation have resulted in excellent outcomes while limiting impatient resource utilization. 3. Screening for appropriate outpatient candidates, in addition to patient education, has been demonstrated to positively influence outcomes. PMID- 7473794 TI - Advances in vitreoretinal surgery: macular hole repair and perfluorocarbon liquids. AB - 1. Idiopathic macular hole was once thought to be an untreatable disorder. During the past several years, surgical techniques have evolved that allow for closure of the macular hole, with restoration of vision in many cases. 2. Perfluorocarbon liquids are an important class of compounds that are under investigation for use in vitreoretinal surgery. The use of these compounds facilitates the performance of intraocular surgical maneuvers in certain difficult-to-manage vitreoretinal disorders. These substances have a high specific gravity and are able to displace water within the eye, facilitating reattachment of the retina in certain types of retinal detachments such as those associated with proliferative vitreoretinopathy and giant retinal tears. PFCLs also aid in the removal of intravitreal lens fragments or foreign bodies and in the repositioning or removal of dislocated intraocular lenses. 3. Ophthalmic nurses and technicians can play an important role in initial screening for these patients. Detection of symptoms may lead to early diagnosis of idiopathic macular hole and subsequent treatment. Patient education emphasizing topics such as the importance of strict postoperative face down head positioning may be critical for successful surgery. PMID- 7473795 TI - Evaluating diplopia with the Maddox rod, Risley's prism, and red glass. AB - 1. Using Maddox rod, Risley's prism, and red glass techniques can provide the doctor with the information needed to diagnose the reasons for diplopia. 2. The Maddox rod and Risley's prism help to determine misalignment in primary position. The red glass test provides valuable information on the type of muscle problems in all other fields of gaze. 3. Both tests should be used for any patient complaining about recent symptoms of diplopia. PMID- 7473796 TI - What's your diagnosis? Partial right third nerve palsy with retinal pathology. PMID- 7473797 TI - Contact lenses: where do we go from here? PMID- 7473798 TI - Treatment with thyrotropin-releasing hormone (TRH) in patients with traumatic spinal cord injuries. AB - Numerous preclinical studies have demonstrated that posttraumatic treatment of spinal cord injury (SCI) with thyrotropin-releasing hormone (TRH) or TRH analogs improves long-term behavioral recovery. The purpose of the present study is to provide preliminary data regarding the safety and potential efficacy of TRH in patients with acute SCI. A total of 20 patients with SCI were classified by clinical examination into complete and incomplete injury groups within 12 h of trauma and randomly assigned in double-blinded fashion to treatment with either TRH (0.2 mg/kg intravenous bolus followed by 0.2 mg/kg/h infusion over 6 h) or vehicle (equal volume physiological saline) placebo. A variety of physiological variables were followed during treatment. Clinical examination included motor and sensory testing, as well as assigning a Sunnybrook score based upon level of function. Patients were examined at 24 h, 72 h, 1 week, 1 month, 4 months, and 12 months after injury. TRH infusions were well tolerated. There appeared to be no discernible treatment effect in patients with complete injuries although data were available from only six such patients at 4 months. For the incomplete injury group, a total of 6 treated and 5 placebo patients had 4-month evaluations. TRH treatment was associated with significantly higher motor, sensory, and Sunnybrook scores than placebo treatment. Because of patients lost to subsequent follow-up, 12-month data were not highly informative. These observations must be interpreted with considerable caution because of the small patient numbers, but together with extensive animal studies they support the need for a larger multicenter clinical trial of TRH. PMID- 7473799 TI - Lack of effect of postinjury treatment with methylprednisolone or tirilazad mesylate on the increase in eicosanoid levels in the acutely injured cat spinal cord. AB - Methylprednisolone (MP) improves motor recovery in spinal cord-injured patients when administered in a 24 h intensive high dose regimen beginning within 8 h after spinal cord injury (SCI). The rationale for this regimen has been based upon the need for high doses (i.e., 30 mg/kg initial IV dose) to inhibit posttraumatic lipid peroxidation (LP) in the injured spinal segment. However, injury also triggers the immediate calcium-mediated activation of phospholipase A2 (PLA2), the release of arachidonic acid, and the enzymatic formation of potentially deleterious prostaglandins (PGE2 alpha, PGE2), thromboxane A2 (TXA2), and leukotrienes (LTs). Thus, in view of the glucocorticoid receptor-mediated inhibition of PLA2 that underlies much of MP's antiinflammatory actions, an additional neuroprotective mechanism may relate to an inhibition of eicosanoid formation. Using the cat spinal cord compression model (180g x 5 min at L3; Na pentobarbitol anesthesia), we examined whether 30 min postinjury dosing with MP (30 mg/kg IV) could attenuate spinal tissue eicosanoid levels measured by enzyme immunoassay at 1 h (Experiment 1). Pial blood flow was measured over the dorsal columns at the injury site using laser doppler flowmetry to monitor posttraumatic hyperperfusion as an index of the microvascular pathophysiology of acute SCI. In vehicle treated animals at 1 h postinjury, there was a significant increase in the tissue levels of PGF2 alpha (+290%), PGE2 (+260%), TXB2 (stable analog of TXA2, +126%), and LTB4 (+73%) in comparison to sham, uninjured animals. However, 6-keto-PGF1 alpha (stable analog of prostacyclin or PGI2) and LTC4 did not increase. Methylprednisolone did not reduce the increase in eicosanoid production. In the case of LTB4 and LTC4, MP actually increased the levels further. In addition, we examined the effects of a double dose MP regimen (30 mg/kg IV at 30 min plus 15 mg/kg IV at 2.5 h postinjury) on spinal cord eicosanoid levels at 4 h postinjury (Experiment 2). At 4 h postinjury, significant increases in PGF2 alpha, PGE2, TXB2, and 6-keto-PGF1 alpha were observed, and with the exception of PGE2, no MP attenuation of the increased eicosanoids was seen. These results fail to provide evidence that postinjury administration of high dose MP exerts a significant anti-PLA2 action. On the other hand, MP effectively inhibited secondary spinal cord pial hyperperfusion, which is believed to be largely mediated by free radical-lipid peroxidative mechanisms. Thus, it seems likely that the protective action of MP on the acute microvascular pathophysiology of SCI is mediated by its well-documented effects on posttraumatic LP.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 7473800 TI - Immunolocalization of endothelin in the traumatized spinal cord: relationship to blood-spinal cord barrier breakdown. AB - The purpose of this study was to determine the relationship between endothelin-1 (ET-1), a prominent vasoactive agent, and the breakdown of the blood-spinal cord barrier along the axis of the cord after a moderate spinal cord injury. In the first study rats (n = 10) were euthanized 24 h after spinal cord injury and compared to sham (n = 5) and unoperated (n = 10) controls. Endothelin and immunoglobulins (IgG) were immunolocalized in adjacent sections of spinal cord using semiquantitative immunocytochemical techniques. In the second study animals were pretreated with the endothelin antagonist Bosentan (n = 6) or vehicle (n = 6) prior to spinal cord injury. Animals were euthanized at 24 h postinjury. Ten minutes prior to euthanasia animals were given horseradish peroxidase (HRP) intravenously. After perfusion fixation sections of cord were prepared for quantitative HRP histochemistry. After spinal cord injury there was enhanced staining for endothelin along the axis of the cord that correlated with the anatomical pattern of barrier breakdown to IgG. In those animals that were pretreated with Bosentan, there was a significant reduction in barrier breakdown along the axis of the injured cord as compared to those animals that received vehicle only. Taken together, this data implicate involvement of endothelin in the axial pattern of barrier breakdown after spinal cord injury. PMID- 7473801 TI - Changes of beta-amyloid precursor protein after compression trauma to the spinal cord: an experimental study in the rat using immunohistochemistry. AB - We evaluated by immunohistochemistry the changes of beta-amyloid precursor protein (beta APP) and beta-amyloid peptide (beta A) in the spinal cord of rats with compression injury at Th8-9 of mild, moderate, and severe degrees. The spinal cord of normal rats and animals with laminectomy revealed immunoreactivity to beta APP in nerve cell bodies, the initial part of a few axons of the gray matter, and in scattered glial cells. At 4 h after compression, beta APP immunoreactivity occurred in a few swollen axons of the longitudinal tracts; such beta APP-immunoreactive axons remained throughout the experimental period of 9 days. The number of immunoreactive axons and the intensity of their immunoreactivity were increased in rats with moderate and severe compression. The caudal Th10 segment exhibited more pronounced accumulation of beta APP immunoreactivity than the cranial Th segment. There was no evidence of beta A accumulation after compression injury. In conclusion, there is a rapidly occurring, long-lasting accumulation of immunoreactive beta-amyloid precursor protein after compression injury of rat spinal cord. This accumulation is related to the degree of impact to the cord. PMID- 7473802 TI - The effect of the glycine site-specific N-methyl-D-aspartate antagonist ACEA1021 on ischemic brain damage caused by acute subdural hematoma in the rat. AB - Acute subdural hematoma (SDH) complicates about 20% of severely head-injured patients, and death and severe disability frequently result, yet over half of these patients may have been conscious, at some time after injury, implying secondary mechanisms of brain damage. Drugs that block the "excitotoxic" effects of glutamate at the N-methyl-D-aspartate (NMDA) receptor have generally been effective in reducing ischemic brain damage associated with SDH in animal models, yet these agents all appear to be associated with major behavioral side effects, in conscious patients, at neuroprotective doses. We therefore evaluated the effects of treatment with a novel antagonist for the glycine binding site of the NMDA receptor (ACEA1021) upon ischemic brain damage, in the rat SDH model. ACEA1021 may be free of psychomotor effects, and may thus permit high dose therapy in conscious trauma and stroke patients. SDH was produced by the slow injection of 0.4 mL autologous blood into the subdural space overlying the parietal cortex. brain damage was assessed histologically at 8 coronal planes, in animals sacrificed 4 h after induction of hematoma. Both pre- and posttreatment with ACEA1021 significantly reduced hemispheric ischemic damage produced by SDH. The magnitude of neuroprotection with this compound (26 to 39% reduction in infarct size) is similar to other NMDA antagonists, and the robust posttreatment effect implies that human studies with this compound should be performed in head injured patients, subject to completion of toxicology testing. PMID- 7473803 TI - Posttraumatic brain hypothermia provides protection from sensorimotor and cognitive behavioral deficits. AB - The purpose of this study was to determine the degree of sensorimotor and cognitive protection conferred by posttraumatic brain hypothermia. Baseline measurements were taken on sensorimotor tasks involving forelimb placing and beam walking, as well as on a spatial navigational task utilizing the water maze. Twenty-four hours after the last baseline measurements, normothermic (37 degrees C) animals were subjected to a fluid percussion pulse (1.9-2.4 atm) over the right parietal sensorimotor cortex. Following trauma, brain temperature was maintained for 3 h at either normothermic (37 degrees C, group TBI-N, n = 12) or hypothermic levels (30 degrees C, group TBI-H, n = 11). Shams (n = 10) underwent all surgical procedures including posttraumatic brain injury (TBI) temperature manipulation, but were not subjected to the fluid percussive pulse. Beam-walking and forelimb placing measures were begun 24 h post-TBI and continued for 2 weeks. Animals were tested on the water maze task for 2 days beginning 24 h post-TBI. TBI produced substantial deficits in contralateral limb placing, which recovered over approximately one week. Hypothermia provided partial protection from these deficits, with TBI-H animals exhibiting intermediate scores that differed from both sham and TBI-N animals (p < 0.03). In the water maze, there was a distinction between groups in the ability to navigate 48 h after TBI. TBI-N animals performed significantly worse than sham and TBI-H animals (both p < 0.01), whereas there was no significant difference between the scores of sham and TBI-H animals. The present data demonstrate that moderate postinjury brain hypothermia can provide protection from sensorimotor and cognitive behavioral deficits as well as neuropathology in a model of traumatic brain injury associated with early neuronal and microvascular injury. PMID- 7473804 TI - Expression of growth inhibitory factor mRNA following cortical injury in rat. AB - Growth inhibitory factor (GIF) inhibits survival and neurite formation of cortical neurons in vitro and is found abundantly in the normal human brain. The role of GIF is still obscure, although it is reported to decrease in the brain in Alzheimer's disease. We examined changes in GIF mRNA expression in a rat cortical ablation model with the aid of an in situ hybridization technique. In sham operated animals, the GIF mRNA was expressed consistently in the cerebral cortex, hippocampus, and thalamus. One day after cortical ablation of the left somatosensory cortex, the expression tended to decrease in the cortex ipsilateral to the injury. Four days after surgery, it increased markedly in the affected cortex and thereafter returned to the level of the control animals except for the area surrounding the injury, where GIF mRNA again increased 2 to 3 weeks after ablation. The transient increase in GIF mRNA expression may reflect efforts to inhibit excessive sprouting of neurites. We also studied the effect of topically applied basic fibroblast growth factor (bFGF), which has a range of neurotrophic effects, on GIF mRNA expression. Topically applied bFGF enhanced the suppression of GIF at 1 day after surgery, though it did not affect the subsequent response. GIF can therefore be assumed to affect the outgrowth of injured neurites and might play a major role in maintenance of the neuronal network in cooperation with other trophic factors. Modification of these factors may be the key to improve neuronal damage after injury. PMID- 7473805 TI - Impaired gustatory neophobia following traumatic brain injury in rats. AB - To investigate the function of the amygdala following traumatic brain injury (TBI), rats were tested on a gustatory neophobia task that is sensitive to amygdala and hippocampal damage. Rats were either injured at a moderate level of fluid percussion injury (2.1 atm) or surgically prepared but not injured (sham injury). Seven days after injury (n = 8) or sham injury (n = 9), rats were habituated to the testing chamber without food items present for 30 min. All rats were then food deprived. Twenty-four hours later, rats were placed in the testing chamber for 30 min and allowed to eat freely from four dishes of different foods: rat chow, raisins, potatoes, and cookies. Results showed that injured and sham injured rats did not differ in their ability to find hidden food, suggesting that TBI does not produce an enduring impairment of olfaction. There was also no difference in the total amount of food eaten between injured and sham groups (p > 0.05). The percentage of each type of food consumed did differ between the two groups with sham controls consuming more familiar food (rat chow) compared to the unfamiliar foods (p < 0.01). The injured animals distributed their eating evenly among the four foods with no particular preference for any one food (p < 0.05). This pattern of eating behavior in injured animals is similar to animals that have lesions to both the hippocampus and amygdala (Sutherland and McDonald, 1990). Therefore, the results of this experiment suggest that, in addition to the hippocampus, the amygdala may also contribute to the behavioral changes observed following TBI. PMID- 7473806 TI - The blood-brain barrier disruption to circulating proteins in the early period after fluid percussion brain injury in rats. AB - Breakdown of the blood-brain barrier (BBB) immediately after traumatic brain injury is not clearly understood. In the present study we focused on the integrity of the BBB to circulating proteins within the first hour after injury. For this purpose, vascular permeability to endogenous albumin and to the exogenous protein tracer horseradish peroxidase (HRP) was examined after a lateral fluid percussion brain injury in rats. Albumin was immunolocalized in brain sections at 3 and 60 min after impact. This distribution was compared with the histochemical localization of HRP given before impact at the same time points. In a separate experiment HRP was given prior to sacrifice to determine the time course for the barrier disruption. Permeability to this protein was assessed at 13, 30, and 60 min after impact. Prominent extravasation of albumin occurred within 3 min of injury and was present in multiple foci within the injured hemisphere. At 60 min the extravasated albumin was present in the same sites, where it was widely distributed. Throughout the related brain parenchyma, little difference was found between the extravascular distribution of albumin and HRP. In the delayed administration paradigm breakdown of the BBB was noted in the impact site, hemorrhagic site in the deep cortical layer, hippocampus, thalamus, and midbrain at 13 min after injury. This injured barrier was restored in most regions by 30 min. However, the impact site and hemorrhagic site remained permeable up to 60 min postinjury. In addition, newly developed barrier disruption to HRP occurred in the parasagittal cortex at 30 and 60 min. In conclusion, widespread breakdown of the BBB to circulating proteins occurred within a few minutes after traumatic brain injury. The time course for this barrier disruption is characterized by three different patterns: (1) transient, (2) prolonged, and (3) delayed opening. This variation in the development of barrier disruption may be related to the secondary barrier failure as well as the primary opening after injury. PMID- 7473808 TI - 3rd International Neurotrauma Symposium. Toronto, Ontario, Canada, July 22-27, 1995. Abstracts. PMID- 7473807 TI - A new model for rapid stretch-induced injury of cells in culture: characterization of the model using astrocytes. AB - The purpose of this study was to develop a simple, reproducible model for examining the morphologic, physiologic, and biochemical consequences of stretch induced injury on tissue-cultured cells of brain origin. Rat cortical astrocytes from 1- to 2-day-old rats were cultured to confluency in commercially available 25-mm-diameter tissue culture wells with a 2-mm-thick flexible silastic bottom. A cell injury controller was used to produce a closed system and exert a rapid positive pressure of known amplitude (psi) and duration (msec). The deformation of the membrane, and thus the stretch of the cells growing on the membrane, was proportional to the amplitude and duration of the air pressure pulse. Extent of cell injury was qualitatively assessed by light and electron microscopy and quantitatively assessed by nuclear uptake of the fluorescent dye propidium iodide, which is excluded from cells with intact membranes. Lactate dehydrogenase (LDH) enzyme release was measured spectrophotometrically. Cell injury was found to be proportional to the extent of the silastic membrane deformation. Increasing cell stretch caused mitochondrial swelling and vacuolization as well as disruption of glial filaments. Stretching also caused increased dye uptake, with maximum dye uptake occurring with a 50 msec pressure pulse duration, whereas deformations produced over longer periods of time (seconds) caused little dye uptake. With increasing postinjury survival fewer cells took up dye, implying cell repair. LDH release was also proportional to the amplitude of cell stretch, with maximum release occurring within 2 h of injury. In summary we have developed a simple, reproducible model to produce graded, strain-related injuries in cultured cells. Our continuing experiments suggest that this model can be used to study the biochemistry and physiology of injury as well as serve as a tool to examine the efficacy of therapeutic agents. PMID- 7473809 TI - Retinoids and apoptosis: implications for cancer chemoprevention and therapy. PMID- 7473810 TI - A new generation of monoclonal antibodies arrives at the clinic. PMID- 7473811 TI - Chemotherapy's benefits bring hope of progress for NSCLC. PMID- 7473812 TI - Probing for DNA clues for cancer diagnosis, treatment, and beyond. PMID- 7473813 TI - Panel endorses behavioral therapy for cancer pain. PMID- 7473814 TI - Local control following breast-conserving surgery for invasive cancer: results of clinical trials. AB - Prospective, randomized clinical trials have demonstrated that the alternatives of mastectomy or conservative surgery plus radiation therapy provide equivalent survival for patients with invasive breast cancer. The identification of a subset of women who could undergo conservative surgery without radiotherapy would avoid the costs, inconvenience, and complications of radiotherapy and is an important research goal. Four randomized trials comparing conservative surgery alone with conservative surgery plus radiotherapy have demonstrated an average reduction in the risk of disease recurrence in the breast of 84% with the use of radiotherapy. No significant differences in survival have been observed, although the available studies lack sufficient numbers of patients to demonstrate a potential small, but clinically important, survival advantage for patients treated with radiotherapy. Subset analysis in the randomized trials and prospective studies of highly selected patients have failed to consistently identify a group of patients who do not benefit from radiation therapy. Any recurrence of breast cancer is psychologically devastating, and fewer than one half of the patients who have had disease recurrence after conservative surgery alone have undergone further breast conserving treatment. At present, a group of patients who do not require radiotherapy has not been reproducibly identified, and radiotherapy should remain a part of breast-conserving therapy for invasive carcinoma. PMID- 7473815 TI - Growth inhibition and induction of apoptosis by fenretinide in small-cell lung cancer cell lines. AB - BACKGROUND: Lung cancer is the major cause of cancer-related death in the United States, with small-cell lung cancer (SCLC) constituting approximately 20% of all cases of lung cancer. Numerous epidemiologic and molecular studies have suggested that alterations in retinoid-signaling pathways play a role in the pathogenesis of lung cancer. Fenretinide [N-(4-hydroxyphenyl)retinamide; HPR] is a synthetic retinoid with minimal toxicity and favorable pharmacokinetics during long-term administration to patients in clinical trials. PURPOSE: The aim of this investigation was to study the effect of HPR on the growth of SCLC cells in vitro. METHODS: Seven SCLC cell lines (NCI-H69, NCI-H82, NCI-H146, NCI-H209, NCI H345, NCI-H446, and NCI-H510A) were exposed continuously to a broad range of concentrations of HPR or all-trans-retinoic acid (RA), and cell viability was determined on day 3 and day 7 by the trypan blue dye exclusion assay. The growth of these cells was compared with that of control vehicle-treated cells to determine survival fraction and the dose resulting in a 50% inhibition of growth when compared with growth of control cells (IC50). The induction of apoptosis was evaluated by fluorescent microscopy, DNA content analysis, and a terminal deoxyribonucleotidyl transferase-based assay that labels 3'-hydroxyl ends of DNA fragments (TUNEL assay) combined with flow cytometric analysis. RESULTS: HPR inhibited growth of a panel of SCLC cell lines at IC50 values that ranged from 0.1 to 3.0 microM (concentrations that are clinically achievable). In all cell lines tested, HPR was a more potent growth inhibitor than RA. By use of fluorescent microscopy, HPR was found to induce morphologic changes consistent with apoptosis in NCI-H82 SCLC cells, including cellular shrinkage, chromatin condensation, and nuclear fragmentation. Flow cytometric analysis revealed decreased DNA content, and TUNEL assay showed increased digoxigenin-uridine triphosphate incorporation in HPR-treated NCI-H82 SCLC cells; these findings are consistent with the induction of apoptosis. CONCLUSIONS: HPR inhibited the in vitro growth of SCLC cells. In NCI-H82 cells, HPR inhibited growth via the induction of apoptosis. PMID- 7473817 TI - Determinants of black/white differences in colon cancer survival. AB - BACKGROUND: Blacks have lower survival rates for colon cancer than whites, possibly related to more advanced stages of disease at diagnosis and to socioeconomic differences between blacks and whites. While the black/white difference in colon cancer survival is well documented, the few studies that have investigated this difference have been limited by the modest number and type of explanatory factors that were considered. PURPOSE: We analyzed data from the National Cancer Institute Black/White Cancer Survival Study to determine 1) what characteristics might contribute to the racial difference in colon cancer survival and 2) if a survival disparity remained between black and white patients after adjustment was made for these characteristics. METHODS: This prospective study included 454 blacks and a stratified random sample of 521 whites, aged 20 79 years, with cancer of the colon diagnosed from January 1, 1985, through December 31, 1986, and who were residents of the metropolitan areas of Atlanta, New Orleans, and San Francisco/Oakland. Follow-up was truncated on December 31, 1990. Cox proportional hazards regression was used to estimate the death rate among blacks relative to that among whites after adjustment for potential explanatory factors, including sociodemographic factors, concurrent (comorbid) medical conditions, stage at diagnosis, tumor characteristics, and treatment. All P values were calculated from two-tailed tests of statistical significance. RESULTS: After adjustment for age, sex, and geographic area, the black-to-white mortality hazard ratio (HR) was 1.5 (95% confidence interval [CI] = 1.2-1.9), indicating that the risk of death among black patients was 50% higher than that among white patients. Further adjustment for stage reduced the excess cancer mortality to 20% (HR = 1.2; 95% CI = 1.0-1.5), decreasing the overall racial difference in excess mortality from 50% to 20% or to a 60% reduction in excess mortality. Although adjustment for poverty reduced the excess mortality by 20%, adjusting for both stage and poverty did not further reduce the racial difference. Among patients with stages II and III disease, blacks had lower survival rates than whites (HR = 1.8; 95% CI = 1.0-3.1 and HR - 1.5; 95% CI = 1.0 2.3, respectively). Among those patients with metastatic disease (stage IV), survival was similar for whites and blacks. CONCLUSIONS: Stage at diagnosis accounted for more than half of the excess colon cancer mortality observed among blacks. Poverty and other socioeconomic conditions, general health status, tumor characteristics, and general patterns of treatment did not further explain the remaining survival disadvantage among blacks. IMPLICATIONS: Because the racial disparity was confined to earlier stages, future studies should investigate whether blacks have more advanced disease at diagnosis and whether less aggressive treatment is provided because of understanding. PMID- 7473816 TI - Proportion of breast cancer cases in the United States explained by well established risk factors. AB - BACKGROUND: Few estimates of the fraction of cases of breast cancer attributable to recognized risk factors have been published. All estimates are based on selected groups, making their generalizability to the U.S. population uncertain. PURPOSE: Our goal was to estimate the fraction of breast cancer cases in the United States attributable to well-established risk factors (i.e., later age at first birth, nulliparity, higher family income, and first-degree family history of breast cancer), using data from the first National Health and Nutrition Examination Survey (NHANES I) Epidemiologic Follow-up Study (NHEFS), the survey and follow-up of a probability sample of the U.S. population. METHODS: From a cohort of 7508 female participants surveyed in the early 1970s, and followed up between 1982 and 1984 and again in 1987, 193 breast cancer cases were accrued for study. We calculated incidence rates, relative risks (RRs), and population attributable risks (PARs) for breast cancer risk factors and extended our results to the U.S. female population by using sample weights from the NHANES I survey. RESULTS: Our PAR estimates suggest that later age at first birth and nulliparity accounted for a large fraction of U.S. breast cancer cases, 29.5% (95% confidence interval [CI] = 5.6%-53.3%); higher income contributed 18.9% (95% CI = -4.3% to 42.1%), and family history of breast cancer accounted for 9.1% (95% CI = 3.0% 15.2%). Taken together, these well-established risk factors accounted for approximately 47% (95% CI = 17%-77%) of breast cancer cases in the NHEFS cohort and about 41% (95% CI = 2%-80%) in the U.S. population. CONCLUSIONS: The RRs for most of these risk factors were modest, but their prevalence as a group was high, leading to estimates that suggest that a substantial proportion of breast cancer cases in the United States are explained by well-established risk factors. IMPLICATIONS: Elucidation of the determinants underlying recognized factors and study of other factors that may confer risk or protection are needed in efforts to advance understanding of breast cancer etiology and to aid in devising strategies for prevention. PMID- 7473818 TI - Characterization of extensive genetic alterations in ductal carcinoma in situ by fluorescence in situ hybridization and molecular analysis. AB - BACKGROUND: The molecular genetic analysis of invasive breast cancer has identified breast cancer as a genetically complex disease. Ductal carcinoma in situ (DCIS) is thought to represent a preinvasive step in breast cancer progression, yet we know little about its biologic behavior or the genetic alterations present. Because of the increasing diagnosis of DCIS by mammography screening and the debate over how DCIS should be managed, there is a clear need to define the molecular events underlying the development of DCIS. PURPOSE: Our purpose was to identify patterns of genetic alterations in DCIS. METHODS: A group of 30 formalin-fixed, paraffin-embedded blocks of tissue collected from 1987 through 1989 from 21 patients with DCIS was studied. Chromosomal imbalances were determined by interphase cytogenetic analysis using the fluorescence in situ hybridization (FISH) technique. DNA probes were used that recognize chromosome specific repetitive sequence loci at the centromeres of chromosomes 1, 3, 4, 6, 7, 8, 9, 10, 11, 16, 17, and 18. FISH was also used to detect ERBB2 gene amplification in DCIS. To complement the FISH studies, microsatellite analysis of markers near the BRCA1 region of chromosome 17 was done on tissue microdissected from multiple areas of DCIS. Chromosomal imbalances were determined by comparisons of chromosomal indices (total number of hybridization spots per total number of nuclei counted) of normal and DCIS tissue, using the two-sided Mann Whitney test. RESULTS: Using FISH, we have identified patterns of DNA loss and gain of certain chromosome-specific centromeric markers in DCIS. We observed frequent gains of markers on chromosomes 3, 10, and 17 as well as loss of chromosome 18-specific centromeric sequences. ERBB2 gene amplification was detected in tumors from four of 15 patients studied and was clearly limited to the tumor cells within the ducts. Because of the availability of topologically distinct regions of tumors from individuals, we were able to show that paired tumor specimens from individuals share genetic alterations and also have unique ones, suggesting clonal diversity within tumors. The combination of FISH and microsatellite analyses suggested that alterations in chromosome 17 may be quite complex; three of five patients whose samples were analyzed had allelic imbalance at markers on the long arm of chromosome 17. CONCLUSIONS: FISH and microsatellite analyses are useful in detecting extensive genetic alterations in DCIS. Examinations of DCIS tissue using these techniques have identified chromosomes 1, 3, 10, 16, 17, and 18 as candidate sites worthy of immediate study. IMPLICATIONS: This approach may give direction to future research aimed at precisely mapping loci altered in DCIS and help in understanding the biologic events associated with tumor progression or recurrence. PMID- 7473819 TI - Preferential association of human papillomavirus with high-grade histologic variants of penile-invasive squamous cell carcinoma. AB - BACKGROUND: Human papillomavirus (HPV) is causally associated with cervical squamous cell carcinoma (SCC) and its precursor lesions. By analogy, HPV is believed to play a role in penile cancer through progression of HPV-associated penile squamous intraepithelial lesions (SIL). HPV DNA has been reported to be present in 100% of high-grade penile SIL, but the percentage of invasive or infiltrating penile SCC that was positive for HPV DNA has varied from study to study (positivity values ranging from 32% to 82%). PURPOSE: To ascertain whether HPV is associated with penile cancer, we used a polymerase chain reaction (PCR) based assay to test specimens of penile SCC for the presence of HPV DNA. METHODS: A total of 117 formalin-fixed, paraffin-embedded specimens of penile cancer from an equal number of patients who had been diagnosed either at the Memorial Sloan Kettering Cancer Research Center in New York City between 1964 and 1992 or the Universidad Nacional de Asuncion in Paraguay between 1980 and 1992 were analyzed. Specimens were examined without prior knowledge of the histology of the lesions. Methods were used that minimized sample contamination, thus avoiding false positive results. PCR and Southern blot analyses were used to determine HPV type. The presence of HPV DNA was studied for association with the tumor properties histopathology, growth pattern, tumor grade, regional lymph node status, and anatomic location. Two-sided statistical tests were used to determine P values. RESULTS: HPV DNA was detected in 26 (22.2%) of 117 specimens. In 23 (88.5%) of the 26 HPV-positive specimens, HPV type 16 (only) was identified. HPV DNA was frequently associated with SCC in areas showing basaloid and/or warty changes (nine [47.4%] of 19 specimens were HPV positive; P = .0125). More highly significant was the association of virus with basaloid SCC (nine [75%] of 12 specimens were HPV positive; P = .0005). However, HPV was not found to be associated with typical SCC of the penis (five [11.1%] of 45 specimens were HPV positive). Virus DNA was more often associated with high-grade tumors (P = .0278) exhibiting aggressive growth (P = .0382) localized to the glans penis (P = .0324). Stepwise logistic regression analysis revealed that only tumor histopathology was a significant predictor of an HPV association. CONCLUSIONS: The presence of HPV DNA was found to be significantly associated only with those penile SCC exhibiting basaloid changes. Furthermore, HPV DNA sequences tended to be associated with higher grade and more aggressive tumor localized to the glans penis. The low frequency of HPV in penile SCC implies that only a small proportion of these cancers arise from HPV-associated penile SIL. PMID- 7473820 TI - Randomized trial of adjuvant human interferon gamma versus observation in high risk cutaneous melanoma: a Southwest Oncology Group study. PMID- 7473821 TI - Re: Inbred strains of laboratory animals: superior to outbred mice? PMID- 7473823 TI - Minimal requirements for characterization of cytokine gene-transduced tumor cells: a proposal. PMID- 7473822 TI - Re: Macrophage colony-stimulating factor complementary DNA: a candidate for gene therapy in metastatic melanoma. PMID- 7473824 TI - Helicobacter pylori and gastric atrophy--cancer paradoxes. PMID- 7473825 TI - Diet and colorectal cancer: still an open question. PMID- 7473827 TI - Monoclonal antibodies progress to the next generation. PMID- 7473826 TI - Does U.S. lag behind Europe in organ-conserving cancer treatment? PMID- 7473828 TI - Whatever happened to beta carotene? PMID- 7473829 TI - The trials of olestra: is a fat-free lunch too costly? PMID- 7473830 TI - Prostate cancer meeting confronts controversies. PMID- 7473831 TI - Recruiting minorities into clinical trials: toward a participant-friendly system. AB - The purpose of this review is to describe the state of the art in recruiting participants for clinical trials designed to test new methods of treatment or disease prevention. The ultimate objective of this review is to provide a summary of key issues in recruiting diverse populations into clinical trials, particularly ethnic and racial minorities. An overview of general issues related to clinical trial participation is followed by a detailed discussion of specific issues that must be addressed by investigators as they recruit minority populations for clinical trial. To date, the majority of clinical trials have included a limited segment of the U.S. population: middle-class, married white males. These trials have faced many problems in their efforts to recruit participants, including barriers to physician participation, barriers to subject participation, barriers to planning and implementing effective recruitment strategies, and costs of the recruitment phase of clinical trials. Following this general introduction is a discussion of the definition of diverse populations. The first step investigators must take as they prepare to recruit study participants is to develop a relevant definition of the subject populations. A detailed review of investigators' experiences in recruiting minorities into clinical trials is presented, including barriers to minority recruitment, barriers inherent in study design, researcher bias, barriers to minority physician participation, as well as strategies for minority recruitment, modifications of study design, and cost issues. Formal evaluation of recruitment strategies has been limited. Most investigators present descriptive reports of their experience in recruiting diverse populations into clinical trials. Research into the issues presented and rigorous testing of specific strategies is needed. A series of steps that are essential to effective clinical trials recruitment of diverse populations is presented. PMID- 7473832 TI - Randomized trial of intake of fat, fiber, and beta carotene to prevent colorectal adenomas. AB - BACKGROUND: Epidemiologic evidence of associations between the high intake of fat and low intake of dietary fiber, beta carotene, and other dietary constituents and the risk of colorectal neoplasia has been inconsistent and has not provided a sufficient basis for recommendations concerning the dietary prevention of large bowel cancer in humans. PURPOSE: We conducted a clinical trial to assess the effects on the incidence of adenomas of reducing dietary fat to 25% of total calories and supplementing the diet with 25 g of wheat bran daily and a capsule of beta carotene (20 mg daily). METHODS: We performed a randomized, partially double-blinded, placebo-controlled factorial trial in which half the patients were assigned to each intervention, resulting in seven intervention groups and one control group. Eligibility criteria included histologic confirmation of at least one colorectal adenoma and confidence expressed by the colonoscopist that all polyps had been removed. Dietary changes were individually initiated and monitored by dietitians and research nurses. At surveillance colonoscopy, the size and location of all polyps were recorded, and their histology was later centrally reviewed. Among 424 patients who were randomly assigned in the trial, 13 were found to be ineligible upon histologic review. Among the remaining 411, complete outcome data were collected from 390 at 24 months and from 306 at 48 months. All P values are from two-sided tests of statistical significance. RESULTS: There was no statistically significant prevention of total new adenomas with any of the interventions. We found a statistically non-significant reduced risk of large adenomas (> or = 10 mm) with the low-fat intervention: At 24 months, the odds ratio (OR) adjusted for potential confounders = 0.4 and 95% confidence interval (CI) = 0.1-1.1; at 48 months, OR = 0.3 and 95% CI = 0.1-1.0. Less and statistically nonsignificant reductions in the risk of large adenomas were found with wheat bran: At 24 months, OR = 0.8 and 95% CI = 0.3-2.2; at 48 months, OR = 0.8 and 95% CI = 0.3-2.5. Patients on the combined intervention of low fat and added wheat bran had zero large adenomas at both 24 and 48 months, a statistically significant finding (P = .03). CONCLUSIONS: Because only small numbers of patients were studied, our finding that the combination of fat reduction and a supplement of wheat bran reduced the incidence of large adenomas in this randomized, controlled trial must be treated with caution. The results do suggest, however, that these interventions may reduce the transition from smaller to larger adenomas, a step that may critically define those adenomas most likely to progress to malignancy. PMID- 7473833 TI - Intake of carotenoids and retinol in relation to risk of prostate cancer. AB - BACKGROUND: Several human studies have observed a direct association between retinol (vitamin A) intake and risk of prostate cancer; other studies have found either an inverse association or no association of intake of beta-carotene (the major provitamin A) with risk of prostate cancer. Data regarding carotenoids other than beta-carotene in relation to prostate cancer risk are sparse. PURPOSE: We concluded a prospective cohort study to examine the relationship between the intake of various carotenoids, retinol, fruits, and vegetables and the risk of prostate cancer. METHODS: Using responses to a validated, semiquantitative food frequency questionnaire mailed to participants in the Health Professionals Follow up Study in 1986, we assessed dietary intake for a 1-year period for a cohort of 47,894 eligible subjects initially free of diagnosed cancer. Follow-up questionnaires were sent to the entire cohort in 1988, 1990, and 1992. We calculated the relative risk (RR) for each of the upper categories of intake of a specific food or nutrient by dividing the incidence rate of prostate cancer among men in each of these categories by the rate among men in the lowest intake level. All P values resulted from two-sided tests. RESULTS: Between 1986 and 1992, 812 new cases of prostate cancer, including 773 non-stage A1 cases, were documented. Intakes of the carotenoids beta-carotene, alpha-carotene, lutein, and beta cryptoxanthin were not associated with risk of non-stage A1 prostate cancer; only lycopene intake was related to lower risk (age- and energy-adjusted RR = 0.79; 95% confidence interval [CI] = 0.64-0.99 for high versus low quintile of intake; P for trend = .04). Of 46 vegetables and fruits or related products, four were significantly associated with lower prostate cancer risk; of the four--tomato sauce (P for trend = .001), tomatoes (P for trend = .03), and pizza (P for trend = .05), but not strawberries--were primary sources of lycopene. Combined intake of tomatoes, tomato sauce, tomato juice, and pizza (which accounted for 82% of lycopene intake) was inversely associated with risk of prostate cancer (multivariate RR = 0.65; 95% CI = 0.44-0.95, for consumption frequency greater than 10 versus less than 1.5 servings per week; P for trend = .01) and advanced (stages C and D) prostate cancers (multivariate RR = 0.47; 95% CI = 0.22-1.00; P for trend = .03). No consistent association was observed for dietary retinol and risk of prostate cancer. CONCLUSIONS: These findings suggest that intake of lycopene or other compounds in tomatoes may reduce prostate cancer risk, but other measured carotenoids are unrelated to risk. IMPLICATIONS: Our findings support recommendations to increase vegetable and fruit consumption to reduce cancer incidence but suggest that tomato-based foods may be especially beneficial regarding prostate cancer risk. PMID- 7473834 TI - Helicobacter pylori and atrophic gastritis: importance of the cagA status. AB - BACKGROUND: Infection with Helicobacter pylori is a major risk factor for the development of atrophic gastritis and gastric cancer. H. pylori strains can differ with respect to the presence of cagA (cytotoxin-associated gene A), a gene encoding a high-molecular-weight immunodominant antigen. H. pylori strains possessing cagA have been associated with enhanced induction of acute gastric inflammation. PURPOSE: We investigated the relationship between cagA status and the development of atrophic gastritis in a cohort of subjects infected with H. pylori. METHODS: Gastrointestinal endoscopy with biopsy sampling was used to study the natural history of gastritis in 58 subjects infected with H. pylori. Biopsy specimens were obtained before and after a mean follow-up period of 11.5 years (range, 10-13 years). The cagA status of each individual was determined at the follow-up visit with the use of an enzyme-linked immunosorbent assay designed to detect the presence of serum immunoglobulin G directed against the CagA protein. Two-sided Fisher's exact tests, McNemar's tests, Student's t tests, and Wilcoxon sum rank tests were used to analyze the data. RESULTS: Twenty-four (41%) of the 58 evaluated subjects had serum antibodies against CagA (i.e., they were cagA positive), and 34 subjects were cagA negative. At the initial visit, moderate to severe atrophic gastritis was observed in eight (33%) of the cagA positive subjects and in six (18%) of the cagA-negative subjects. At that time, positive cagA status and gastric atrophy were not significantly related (P = .22; Fisher's exact test; odds ratio [OR] 2.33; 95% confidence interval [CI] = 0.58 9.65). During follow-up, 16 (36%) of the 44 initially atrophy-negative subjects developed atrophic gastritis (eight [50%] of 16 cagA-positive subjects versus eight [29%] of 28 cagA-negative subjects; P = .20, Fisher's exact test; relative risk [RR] = 1.75; 95% CI = 0.82-3.76). In six of these 16 subjects (five cagA positive versus one cagA negative), atrophic gastritis was accompanied by the development of intestinal metaplasia (i.e., a change in the type of specialized cells present) (P = .02; Fisher's exact test; RR = 9.06; 95% CI = 1.16-71.0). One of the initially atrophy-negative, cagA-positive subjects developed early gastric cancer. Four (29%) of the 14 subjects initially diagnosed with atrophic gastritis showed regression of atrophy during follow-up (one cagA positive and three cagA negative). Therefore, at the end of follow-up, 15 (62%) of the 24 cagA-positive subjects had atrophic gastritis compared with 11 (32%) of the 34 cagA-negative subjects (P = .02; Fisher's exact test; OR = 3.48; 95% CI = 1.02-12.18). CONCLUSION: Infection with cagA-positive H. pylori strains is associated with an increased risk for the eventual development of atrophic gastritis and intestinal metaplasia. PMID- 7473835 TI - Effect of beta-carotene supplementation on indices of colonic cell proliferation. AB - BACKGROUND: Epidemiologic studies have shown that consuming foods containing beta carotene is associated with a decreased incidence of colon cancer. The validity of this association has recently been questioned. It is not known if the rate of colonic cell proliferation differs among individuals with or without a history of colonic polyps or cancer and if proliferation changes in response to beta carotene. PURPOSE: This study was intended to (a) determine whether differences exist in colonic cell proliferation in individuals with and without prior colonic polyps or tumors, (b) demonstrate that beta-carotene accumulates in colonic mucosa following dietary supplementation, and (c) determine whether mucosal beta carotene accumulation influences colonic cell proliferation. METHODS: Subjects were enrolled in the phase I study from June 1991 until February 1994. The participants included 20 individuals (11 males and nine females, aged 62.3 +/- 8.9 years [means +/- SD]) with normal colons (as judged by recent colonoscopy), 40 (24 males and 16 females, aged 59.6 +/- 10.1 years) with a history of colonic polyp(s), and 41 (30 males and 11 females, aged 67.2 +/- 9.7 years) with prior colon cancer. The subjects in the last two groups consumed either 30 mg of beta carotene or placebo each morning for 3 months. This dose of beta-carotene has no known toxic effects, but it can increase the serum level by approximately 10 fold. beta-carotene concentration in serum and colonic tissue was quantitated by high-pressure liquid chromatography in samples collected before and after supplementation with beta-carotene or placebo. Cellular proliferation was assessed on the basis of tissue ornithine decarboxylase activity, urinary polyamine excretion, and proliferating cell nuclear antigen expression. The differences in colonic cell proliferation parameters due to beta-carotene supplementation, within and among different groups, were evaluated by the Wilcoxon matched-pairs signed ranked test and the Mann-Whitney test, respectively. All statistical tests were two-sided. RESULTS: Colonic cell proliferation did not differ in samples obtained from individuals with and without prior colonic polyp(s) or cancer. beta-carotene concentrations in serum and colonic tissue were significantly increased in groups receiving beta-carotene (P < .001). However, cell proliferation did not differ, as judged by any of the three measures, among samples from all experimental groups collected before and after supplementation with beta-carotene. CONCLUSIONS: Dietary supplementation with beta-carotene for a period of 3 months does not alter colonic cell proliferation in individuals with a history of colonic polyps or cancer. IMPLICATIONS: The mechanism by which beta-carotene might reduce colon cancer incidence does not appear to involve or result in a change in cell proliferation in the normal colonic mucosa as studied in individuals with a history of colonic polyps or cancer. PMID- 7473837 TI - Re: Tumor angiogenesis as a prognostic assay for invasive ductal breast carcinoma. PMID- 7473836 TI - Inhibition of growth of human osteosarcomas by antagonists of growth hormone releasing hormone. AB - BACKGROUND: Insulin-like growth factor I (IGF-I) may be involved in the proliferation of human osteosarcomas. Most of the IGF-I found in blood is produced in the liver, where transcription of the IGF-I gene is regulated by growth hormone (GH). Recently, we synthesized various potent antagonists of GH releasing hormone (GH-RH), including [Ibu0, D-Arg2, Phe(4-Cl)6, Abu15, Nle27]hGH RH(1-28)Agm, which is also called MZ-4-71. PURPOSE: We investigated the effects of this antagonist on the growth of the human osteosarcoma cell lines SK-ES-1 and MNNG/HOS, transplanted into nude mice or cultured in vitro. METHODS: Nude male mice bearing SK-ES-1 and MNNG/HOS tumors were treated for 4 and 3 weeks, respectively, with MZ-4-71 administered from osmotic minipumps at a dose of 40 micrograms per animal per day. Tumor volume, tumor weight, and levels of receptors for IGF-I were determined. IGF-I levels in serum, tumor, and liver tissue were measured by radioimmunoassay. In other experiments, tumor-bearing nude mice were treated subcutaneously for 3 weeks with the GH-RH agonist hGH-RH(1 29)NH2 or with MZ-4-71 for 13 days at doses of 50 micrograms per animal per day. Effects of MZ-4-71, hGH-RH(1-29)NH2, and human GH (hGH) on cell proliferation and on the production of IGF-I and cyclic adenosine monophosphate were also evaluated in SK-ES-1 and MNNG/HOS cells in vitro. RESULTS: The growth of SK-ES-1 and MNNG/HOS tumors in nude mice was significantly inhibited by MZ-4-71, as measured by a reduction in tumor volume and weight (all P values < .05). MZ-4-71 treatment of either SK-ES-1 or MNNG/HOS tumor-bearing animals decreased tumor tissue IGF-I levels. The growth of MNNG/HOS xenografts was stimulated by hGH-RH(1-29)NH2 (P < .01). IGF-I levels in serum of tumor-bearing nude mice treated subcutaneously for 13 days with MZ-4-71 were decreased (both P values < .01). High-affinity binding sites for IGF-I were demonstrated on cell membranes of SK-ES-1 and MNNG/HOS tumors. In cell cultures of both osteosarcomas, IGF-I production was stimulated by 25 ng/mL hGH but was not changed by 10 ng/mL hGH-RH(1-29)NH2 or 5 microM MZ-4 71. Incorporation of [3H]thymidine into DNA in SK-ES-1 (but not MNNG/HOS) cells was increased by 25 ng/mL IGF-I (P < .01). The proliferation rate of the two cell lines was not affected by 5-50 ng/mL hGH-RH(1-29)NH2 or 1-80 ng/mL hGH but was suppressed by 10(-6)-10(-5) M MZ-4-71. CONCLUSIONS: Our findings demonstrate that the GH-RH antagonist MZ-4-71 can significantly inhibit the growth of SK-ES-1 and MNNG/HOS osteosarcomas in mice. PMID- 7473838 TI - Re: Tumor angiogenesis as a prognostic assay for invasive ductal breast carcinoma. PMID- 7473839 TI - Re: Tumor angiogenesis as a prognostic assay for invasive ductal breast carcinoma. PMID- 7473840 TI - Re: Detection of a point mutation in NQO1 (DT-diaphorase) in a patient with colon cancer. PMID- 7473841 TI - Active tumor-specific immunization of the bone marrow (progenitor stem cell) donor. PMID- 7473842 TI - Does paclitaxel penetrate into brain tumor tissue? PMID- 7473843 TI - Cooperative research sought by the Prof. N.N. Petrov Research Institute of Oncology, St. Petersburg. PMID- 7473844 TI - AIDS is a multifactorial neuroimmunoendocrine disorder due to energy crisis in the 'Milieu Interieur'. PMID- 7473845 TI - Health care for the 21st century: preserving what's right. PMID- 7473846 TI - Today's physician, tomorrow's world. PMID- 7473847 TI - Neuroepidemiology of vascular and Alzheimer's dementia among African-American women. AB - To better understand risk factors for Alzheimer's disease and vascular dementia, demographic, medical, and other epidemiological factors were compared for 83 African-American women with Alzheimer's disease and 46 with vascular dementia. Overall, the risk-factor profiles for Alzheimer's disease and vascular dementia were similar to those in other studies. However, Alzheimer's patients had a high frequency of hypertension and a relatively high frequency of diabetes mellitus. The presence of such risk factors raises the possibility that there is a vascular component to the dementia in these African-American women with Alzheimer's disease. Neuropathological studies are needed to help answer this question. PMID- 7473848 TI - Hypercalcemia associated with adult T-cell leukemia/lymphoma (ATL). AB - Hypercalcemia is a frequent manifestation of human T-cell lymphotrophic virus type I (HTLV-I)-associated adult T-cell leukemia/lymphoma (ATL). Human T-cell lymphotrophic virus type I infection is endemic in the Caribbean, Japan, Melanesia, and Africa. This article presents two cases of ATL to increase awareness of the disease by primary care physicians. The management of hypercalcemia is discussed. PMID- 7473849 TI - The significance and work-up of minor gastrointestinal bleeding in hospitalized nursing home patients. AB - Twenty-seven consecutive admissions from nursing homes who underwent a gastrointestinal (GI) consult for coffee ground vomitus or occult GI bleeding to evaluate the outcome were reviewed retrospectively to determine whether a GI work up was or would have been useful. There were 15 deaths, all associated with severe infection or respiratory failure. Endoscopy, barium studies, and a history of nonsteroidal anti-inflammatory drug use or peptic ulcer disease did not affect the management or outcome. No patient developed major GI bleeding. When nursing home patients present with coffee ground vomitus or newly found occult blood in the stool, efforts should be made to identify and vigorously treat any acute underlying infection or respiratory failure. Endoscopy is not helpful in this clinical situation. Both the primary care physician and the GI consultant should be aware of these associations and should focus on the underlying etiology. PMID- 7473850 TI - Barriers to treatment for addicted African-American women. AB - This article describes barriers to treatment identified by a sample of substance abusing/addicted women (mostly African-American). A self-administered questionnaire, the Allen Barriers to Treatment Instrument, was submitted to 97 substance-abusing/addicted women throughout the state of Illinois who were not in treatment. Results showed that the barriers most identified by these subjects included responsibility for child care, lack of insurance or money, and community issues. PMID- 7473851 TI - Assessing the risk of gestational diabetes in twin gestation. AB - This study examines the hypothesis that twin gestation is a risk factor for gestational diabetes. In a retrospective analysis, the incidence of gestational diabetes in twin and singleton pregnancies was determined in groups matched for maternal age, weight, and parity. One-hour oral glucose challenge tests (50 g) were used to screen 9185 pregnant women. Gestational diabetes was diagnosed when abnormal screens (> or = 130 mg/dL) were followed by two or more abnormal values on a 3-hour (100 g) glucose tolerance test using National Diabetes Data Group (NDDG) criteria. A twin gestation was identified in 1.5% (138/9185) of the pregnancies. Gestational diabetes was diagnosed in 5.8% (8/138) and 5.4% (439/9047) of the twin and singleton pregnancies, respectively. The incidence of gestational diabetes is similar for singleton and twin gestations. PMID- 7473852 TI - The impact of transrectal ultrasound on stage migration in a predominantly African-American population. AB - An important question left unanswered is whether transrectal ultrasound will result in earlier diagnosis in African Americans. Tumor registry data for 1985 and 1990 for a predominantly African-American population were reviewed to determine whether transrectal ultrasound influenced the stage at diagnosis of prostate cancer. Diagnosis by ultrasound increased from 0% to 60% of cases in those 5 years. Curable diseases (stages A and B) increased from 38% to 57% of cases. It is concluded that transrectal ultrasound can increase the diagnostic yield of potentially curable disease in a predominantly African-American population. PMID- 7473853 TI - End-stage renal disease in patients infected with human immunodeficiency virus: a retrospective review of 38 cases. AB - A retrospective review was conducted to evaluate the influence of risk factors for human immunodeficiency virus (HIV) infection on the outcome of patients with end-stage renal disease (ESRD). The records of all patients seen at Howard University Hospital between February 1984 and July 1994 with a diagnosis of HIV infection were reviewed. Two hundred seventy-eight patients had a diagnosis of renal failure; 38 of these patients developed end-stage renal failure requiring dialysis. Risk factors for HIV infection in these patients were intravenous drug abuse, homosexual behavior, bisexual preference, and blood transfusion. None of these factors consistently influenced the survival of HIV-infected patients with ESRD. PMID- 7473854 TI - Massive lower gastrointestinal bleeding due to 'Dieulafoy's vascular malformation' of the jejunum: case report. AB - Dieulafoy reported three cases of massive gastric hemorrhage due to a dilated submucosal artery in 1898, and since then, more than 100 cases of this gastric vascular malformation have been reported in the literature. These same pathologic lesions are even a rarer occurrence in the small bowel. This article reports a 38 year-old hypotensive male who presented to the hospital after an acute onset of massive lower gastrointestinal hemorrhage; superior mesenteric angiography demonstrated an actively bleeding lesion in a proximal jejunal branch. Intraoperative small bowel endoscopy via an enterotomy demonstrated a 4 mm bleeding submucosal lesion 30 cm distal to the ligament of Treitz. A literature review revealed six other cases of Dieulafoy's vascular malformation that occurred in the small bowel, with the lesions located in the proximal jejunum between 15 cm and 45 cm distal to the ligament of Treitz. The cause of these lesions is unknown. This case demonstrates the importance of preoperative angiography and intraoperative endoscopy when massive lower gastrointestinal hemorrhage is suspected to be from a small bowel source. PMID- 7473855 TI - Crisis in radiology: are radiologists really necessary? PMID- 7473856 TI - Evidence for common binding of acetaminophen and bromobenzene to the 58-kDa acetaminophen-binding protein. AB - Acetaminophen (APAP) toxicity has been closely associated with covalent binding to a cytosolic protein of approximately 58 kDa (58-ABP). To determine if metabolites of other toxicants might also selectively target this protein, studies were conducted with bromobenzene (BrB). Mice were given phenobarbital (80 mg/kg/d x 4 d) and were killed 4 h after challenge with 800 mg BrB/kg. Liver cytosols from BrB-treated or naive mice were incubated with an APAP activating system. Cytosolic fractions were analyzed for APAP binding by Western blotting with anti-APAP antibody. Binding to 58-ABP was selectively decreased in liver cytosol from BrB-treated mice while binding to other targets was minimally affected. Western blotting of the same samples with anti-58-ABP antisera showed that this decrease in binding did not result from diminished 58-ABP content. HPLC analysis of APAP-N-acetyl cysteine conjugate formation in vitro indicates that APAP activation was not altered in the incubates with cytosol from BrB-treated mice. These results suggest that the 58-ABP may be a common target for electrophiles in reactive intermediate toxicity. PMID- 7473857 TI - Induction of cytochrome P-450 in the Norway rat, Rattus norvegicus, following exposure to potential environmental contaminants. AB - Cytochrome P-450 (CYP) induction (consisting of increases in cellular RNA and protein content and associated catalytic activities) occurs predominantly in the liver, but also in small intestine, lung, kidney, and placenta, of Norway rats (Rattus norvegicus) exposed to certain types of potential environmental contaminants. The specific isoform(s) induced in the rat and the magnitudes of the increases observed depend upon the chemical nature of the xenobiotic. For instance, the predominant isoforms induced by nonhalogenated polycyclic aromatic hydrocarbons, such as petroleum derivatives and coal-tar constituents such as the benzopyrenes and the anthracenes, are those of the CYP1A subfamily. Polyhalogenated aromatic hydrocarbons, such as the halogenated dibenzodioxins, dibenzofurans, and biphenyls, may cause the induction of predominantly the CYP1A subfamily, predominantly the CYP2B subfamily, or mixed CYP1A- and CYP2B-type induction, depending upon the halogen substitution pattern. In contrast, the chlorinated hydrocarbon pesticides, such as DDT, dieldrin, chlordane, and mirex, cause almost exclusively the induction of isoforms of the CYP2B (and to a lesser extent the CYP3A) subfamilies. The commonly employed plasticizing agent di-(2 ethylhexyl)phthalate elicits predominantly induction of the CYP4A subfamily. Those xenobiotics that would be expected to be the most pervasive environmental contaminants are typically those that have also been found to cause the most profound CYP induction responses. Such chemicals are extremely lipophilic and tend to accumulate in animal tissues, especially fatty tissues such as the liver. The hepatic CYP induction response to such potential environmental contaminants is typical of the animals' response to lipophilic xenobiotics in general, and serves as a mechanism by which the excretion of such compounds from the body is facilitated. PMID- 7473858 TI - Organochlorine residues in human adipose and hepatic tissues from autopsy sources in northern Italy. AB - Organochlorine compounds have been analyzed in human samples from residents of Genoa, a city in Northern Italy. Twenty-eight specimens of adipose tissue from 17 males and 11 females deceased from accidental causes in March and April 1989 were examined. In 12 of the 17 males, liver tissue specimens were also analyzed. DDE was the major organochlorine pesticide (OCP) found in all human adipose tissues (395 +/- 264 ng/g); additional OCPs found in our series of human subjects with an occurrence greater than 80% included p,p-DDT (64 +/- 31 ng/g), lindane (104 +/- 93 ng/g), and beta-BHC (213 +/- 260 ng/g). Statistical analysis showed positive correlation of DDE and p,p-DDT with age with an annual increase of 7 ng/yr and 0.9 ng/yr, respectively. No statistically significant differences were found for OCPs concentration between sexes. In DDE to sigma DDT (o,p-DDT + p,p-DDT + o,p TDE + p,p-TDE) ratio changed in the last two decades from 0.95 (1966) to 5.2. Significant correlation was found between paired liver-adipose tissues concentrations of DDE and p,p-DDT (1:2 ratio). No correlation was found between age of subjects and the DDE to sigma DDT ratio. This study indicates that the ban on DDT use as a pesticide has been largely effective in Liguria, as supported by the lower concentrations of the compound in youngsters and the increase in the DDE to sigma DDT ratio with time. However, other data, such as the lack of correlation between age of subject and the DDE to sigma DDT ratio, indicate that intake of minimal amounts of DDT may have taken place even recently. This intake could depend on contamination of food imported from abroad or on illegal, or incorrect, use of these compounds in Italy. PMID- 7473859 TI - Rainbow trout hepatic mixed-function oxygenase induction by polychlorinated dibenzo-p-dioxins (PCDDs) as a function of time and tissue concentration. AB - Rainbow trout, dosed orally with 0.060-84 micrograms/kg of [3H]-2,3,7,8 tetrachlorodibenzo-p-dioxin (TCDD), [14C]-1,2,3,4,7,8-hexachlorodibenzo-p-dioxin (HxCDD), or [14C]-1,2,3,4,6,7,8-heptachlorodibenzo-p-dioxin (HpCDD) showed dose dependent increases in hepatic ethoxyresorufin O-deethylase (EROD) activity, up to 250-fold, after 2-16 d. Induction of EROD activity was a sensitive and rapid indicator of exposure to PCDDs. The effects of time after exposure were not dramatic, but generally EROD activity after 2 d was lower than EROD activity after 16 d. Slopes of dose-response curves relating EROD activity to hepatic concentrations of PCDDs significantly decreased and intercepts significantly increased with increasing time of exposure. When fish were grouped by oral doses, only the low doses of TCDD and HpCDD exhibited time-dependent EROD induction, with significantly greater activity 16 d after dosing compared to 2 d. In most cases, hepatic and muscle concentrations of PCDDs did not significantly change over time. Concentrations of PCDDs in liver and muscle accounted for up to 7% of the orally administered dose. Hepatic levels of PCDDs ranged from 20-100 pg/g at the lowest doses to about 1000 pg TCDD/g, 2000 pg HxCDD/g, or 40,000 pg HpCDD/g at the highest doses. Of the PCDD in the liver, approximately one-third to one half was associated with the postmitochondrial supernatant (PMS). PCDD concentrations in muscle did not span as wide a range of concentrations as did PCDD in liver; fish given low doses had 10 pg/g in muscle whereas fish given high doses had several hundred picograms per gram in muscle for all three PCDDs. PMID- 7473860 TI - Ketone potentiation of haloalkane-induced hepato- and nephrotoxicity. II. Implication of monooxygenases. AB - Previous results in Sprague-Dawley rats indicate that acetone (A), methyl ethyl ketone (MEK), and methyl isobutyl ketone (MiBK) pretreatment (3 d, po) at dosages of 6.8 and 13.6 mmol/kg potentiate CCl4 hepatotoxicity and CHCl3 nephrotoxicity, respectively. The potentiation potency profile observed was MiBK > A > MEK for liver and A > MEK > or = MiBK for kidney toxicity (Raymond & Plaa, 1995). In the present study, hepatic and renal microsomes from A-, MEK-, and MiBK-pretreated rats (6.8 or 13.6 mmol/kg) were examined for cytochrome P-450 content, substrate specific monooxygenase activity (aminopyrine and benzphetamine N-demethylase, aniline hydroxylase) and in vitro covalent binding of 14CHCl3 and 14CCl4. Of the three ketones, only MiBK significantly increased P-450 content of liver and renal cortical microsomes. Similarly, 14CCl4 covalent binding under aerobic and anaerobic conditions was significantly increased by MiBK pretreatment only. 14CHCl3 covalent binding by renal cortical microsomes was significantly increased only under aerobic conditions by MiBK pretreatment. MiBK (13.6 mmol/kg) increased (threefold) aminopyrine N-demethylation in both liver and kidney, but only benzphetamine N-demethylation (two-fold, at 6.8 and 13.6 mmol/kg) in liver; A and MEK had no effect on either monooxygenase. All ketones at dosages of 6.8 and 13.6 mmol/kg increased aniline hydroxylation in liver (two-fold) and kidney (fivefold). Comparable profiles for P-450 induction, haloalkane covalent binding, and aminopyrine or benzphetamine N-demethylase activity were observed in liver and kidney microsomes. This profile was consistent with the ketone potentiation potency ranking profile observed in vivo for liver but not kidney injury. These findings affirm the importance of ketone-enhanced bioactivation for potentiation of CCl4 hepatotoxicity but suggest an alternative mechanism for CHCl3 nephrotoxicity. PMID- 7473861 TI - Effects of ozone on the epithelial and inflammatory responses in the airways: role of tumor necrosis factor. AB - We have investigated the possibility that tumor necrosis factor alpha (TNF) plays a role in the increased airway permeability and an inflammatory response following an acute ozone (O3) exposure. Male Sprague-Dawley rats were injected, intraperitoneally, with either rabbit anti-mouse antibody to TNF (anti-TNF) or preimmune rabbit serum, 2 h before a 3-h exposure to O3 or purified air. Permeability, as determined by [99mTc] diethylenetriamine pentaacetate (DTPA) transport, total protein and albumin concentrations in the bronchoalveolar lavage (BAL), and the inflammatory cell response in the BAL were assessed 10 h after the exposure was completed. The O3-exposed group that was injected with anti-TNF showed a significant decrease in permeability to DTPA in comparison to the O3- exposed group injected with preimmune rabbit serum. There was no difference between the anti-TNF group and the purified air group. In contrast, the total protein and albumin levels in the BAL were significantly greater in both of the O3-exposed groups than in the purified air group. The concentrations of protein and albumin in the anti-TNF group did, however, show an attenuating trend when compared to the preimmune O3-exposed group. The polymorphonuclear leukocytes (PMNs) in BAL of the anti-TNF group also showed an attenuating trend when compared to the preimmune O3-exposed group, but both of these O3-exposed groups were significantly greater than the purified air group. Lung sections stained with naphthol AS-D chloroacetate esterase showed an increase in the number of stained PMNs in the anti-TNF group in comparison to the preimmune O3- and air exposed groups. These data suggest that TNF plays a role in the increase in tracheal permeability as determined by DTPA transport, while the contributing role that TNF plays in bronchoalveolar permeability and the inflammatory response seen following an acute exposure to 0.8 ppm O3 is less evident. PMID- 7473862 TI - Mechanisms of hepatic methylmercury uptake. AB - The mechanism by which methylmercury is cleared from hepatic portal blood was examined in isolated rat livers perfused single-pass with Krebs-Henseleit buffer. [203Hg]Methylmercury (0.24-24 microM) was infused over a 30-min interval, followed by a 30-min washout, as a complex with the endogenous ligands L-cysteine (CH3Hg-L-cys), glutathione (CH3Hg-SG), and serum albumin (CH3Hg-albumin), or as a complex with dithiothreitol (CH3Hg-DTT), chloride (CH3HgCl), and the D-enantiomer of cysteine (CH3Hg-D-cys). The sulfhydryl-containing compounds were added at a 10 fold molar excess. When administered as the albumin complex, only a small fraction of the [203Hg]methylmercury was cleared from perfusate (approximately 8%) and excreted into bile (0.7%). Hepatic uptake and biliary excretion of methylmercury was considerably higher for the other complexes: The percent of the dose recovered in liver tissue and bile was, respectively, CH3Hg-albumin, 6.9 and 0.7; CH3Hg-L-cys, 15.7 and 2.3; CH3Hg-D-cys, 27.1 and 2.8; CH3Hg-SG, 17.7 and 2.1; CH3HgCl, 66.5 and 3.2; and CH3Hg-DTT, 70.1 and 19.8. For the dithiothreitol complex, hepatic extraction of methylmercury was nearly complete during single pass perfusion. A comparison of hepatic removal of increasing doses of CH3Hg-L cys and CH3Hg-D-cys revealed little difference in uptake between these two enantiomers. Moreover, the fraction of methylmercury removed was similar when infused at concentrations of 0.24, 2.4, and 24 microM, indicating no saturability of uptake within this dose range. Methylmercury was not hepatotoxic at concentrations up to 24 microM if administered as a mercaptide; however, the chloride complex (CH3HgCl) produced cholestasis and an increase in perfusion pressure at a concentration of only 0.24 microM. These findings indicate that hepatic methylmercury uptake and toxicity are dependent on the chemical form in blood plasma. Uptake was faster when methylmercury was present as a cysteine or glutathione complex, as compared to the albumin complex; however, the lack of stereoselectivity indicates that the uptake process may be relatively unselective. PMID- 7473863 TI - Oral bis(tri-n-butyltin) oxide in pregnant mice. I. Potential influence of maternal behavior on postnatal mortality. AB - Pregnant Swiss mice were treated with 0, 5, 10, 20, and 30 mg/kg body weight of bis(tri-n-butyltin) oxide (TBTO) on d 6-15 of gestation. At birth litters were normalized to eight pups, and postnatal evaluation of pup growth rate and behavioral observations of dams were carried out. Litters were sacrificed on postnatal days (pnd) 7, 14, and 21, to perform hematological analysis, in connection with another study. Dam weight gain was impaired in all the treated groups (except at the lowest dose level) in the late phase of gestation. A high incidence of anticipated or delayed parturitions, without any correlation with fetal mass, was observed in the treated groups. All the treated dams showed a significant increase in resorptions, and a decrease in body weight gain between gestational day (gd) 6 and pnd 1. At birth, only the 20 and 30 mg/kg dose groups showed reduced litter size and reduced pup weight. Body weight gain reduction of pups persisted in wk 1 of life only in the 10 and 20 mg/kg dose groups. In addition, the maternal weight trend was affected during the lactation period in the higher dose groups. Postnatal death rate and growth rate of treated pups were affected by an altered maternal behavior; pups, apparently viable and with normal weight, were found often scattered throughout the cage with signs of wounds, and the percentage of dams that had not built a nest increased in the 10, 20, and 30 mg/kg dose groups. Total absence of parental care was noted in many litters, and many infanticidal events were reported. Our results seem to confirm low TBTO embryofetotoxicity, and strongly support the assumption that TBTO's toxicity to the mother is much stronger than its embryo-fetotoxic potential. Most of the reproductive parameters examined in this study were unaffected in the low-dose group, while some indices, such as gestation length and maternal weight gain between gd 6 and pnd 1, were markedly altered also at the 5 mg/kg dose level and appear to be sensitive parameters in assessing maternal toxicity. PMID- 7473864 TI - Oral bis(tri-n-butyltin) oxide in pregnant mice. II. Alterations in hematological parameters. AB - The effect of in utero TBTO exposure on blood composition in neonates, pups during weaning, and dams in the same period were investigated in mice. The dams were gavaged at dose levels 0, 5, 10, and 20 mg TBTO/kg body weight on gestational days 6-15. At birth, litters were culled to eight pups, and blood analysis was performed on excess pups. On d 7, 14, and 21 the entire litters were sacrificed and blood (WBC, RBC, HGB, HCT, MCV, MCH, MCHC, and differential leukocyte count) of dams and pups was analyzed. Blood analysis of neonates revealed enhanced WBC count and altered red-cell parameters. Increase in WBC persisted until postnatal day (pnd) 21, while red-cell parameters regained normal values on pnd 7. No significant differences were found in thymus and spleen weights in the treated groups. Dams presented no alteration in blood composition or of spleen or thymus weights. The results of this investigation revealed that in utero exposure to TBTO induces a nonspecific alteration of hematological parameters in mice, but further studies are necessary to understand whether our results are either dependent on maternal status or a direct effect of TBTO on embryonic tissue. PMID- 7473865 TI - Interactions of rat red blood cell sulfhydryls with arsenate and arsenite. AB - Arsenic-thiol interactions were investigated by determining changes in rat blood sulfhydryls after exposure to arsenate, As(V), or arsenite, As(III). Incubation with As(V) resulted in time- and dose-dependent depletion of nonprotein sulfhydryls (NPSH), specifically glutathione (GSH). At the highest As(V) concentration (10 mM), significant loss of glutathione was only observed after 3 h of incubation, but by 5 h 0.5 mM As(V) and higher was sufficient to deplete GSH. As(V) was reduced to As(III) at all dose levels, indicating a redox interaction with GSH, but oxidized glutathione (GSSG) was not formed in sufficient quantities to account for losses in GSH. This may be due to formation of another oxidized species such as a protein-mixed-disulfide (ProSSG). Further evidence that glutathione reduces arsenate was obtained by pretreating cells with the sulfhydryl derivatizing agent N-ethylmaleimide (NEM). Removal of thiols with NEM severely inhibited the formation of As(III) in these incubations, indicating that the main pathway for arsenate reduction in red cells is sulfhydryl dependent. As(III) demonstrated a completely different profile of sulfhydryl interaction. Sulfhydryls (NPSH and GSH) were depleted but the losses were primarily accounted for by oxidation to GSSG. As(III) was also a more potent sulfhydryl depleting agent, requiring only 0.1 mM As(III) to significantly reduce GSH after 5 h of incubation. Significant levels of GSSG formed at all doses of As(III). Evidence is presented to suggest that As(III) also formed mixed complexes with protein and glutathione. Samples that were acid precipitated displayed loss of cytosolic glutathione, which could be reversed if NEM was added prior to protein precipitation. Arsenic was detected in high quantities in the protein precipitates, and this was also found to be reversible by NEM treatment. The fact that both GSH depletion and protein binding were reversible by NEM treatment points to formation of a mixed complex of protein, GSH, and As(III), possibly ProS-As-(SG)x. Arsenic affinity chromatography and polyacrylamide gel electrophoresis were used to characterize arsenic binding proteins in red-cell cytosol. The main arsenic binding protein appeared to be hemoglobin. PMID- 7473866 TI - The microbial flora associated with stable endosseous implants. AB - Eleven patients with 36 stable endosseous implants in function for at least six months were selected for culturing in this study. Each patient had been restored with a mandibular implant-borne bar overdenture. The bacterial samples were taken from the superior aspect of the implants after bar and abutment removal. The plaque specimens contained predominantly Gram-positive microflora, although Gram negative species were present in low numbers. This study serves to corroborate previously published reports regarding the characterization of the microflora associated with stable endosseous implants. PMID- 7473867 TI - Will the real implantologist please stand up? PMID- 7473868 TI - Levels of osseointegration of blade-/plate-form implants. AB - The blade-/plate-form implant is the only endosseous implant which has proved to be successful under three different types of integration: fibrous integration, when the layer of fibrous or collagenous tissue interspersed between implant and bone is less than 125 microns; fibro-osseous integration, when there is a combination of thinly defined fibrous connection with 22% or less direct bone contact with the implant; and osseous integration, when there is 23% or more direct bone-to-implant contact, with no evidence of fibrous connective tissue anywhere. The blade implant is an unique versatile device which has passed with success the test of time (some of them being in function for as long as 28 years) and deserves more attention from implantologists around the world. PMID- 7473869 TI - The subperiosteal implant as a viable long-term treatment modality in the severely atrophied mandible: a patient's 40-year case history. AB - Subperiosteal implants have been a treatment modality for atrophied edentulous mandibles for almost 50 years. A case is presented tracing a patient's 40-year history with treatment of his severely atrophied fully edentulous mandible with a subperiosteal implant and removable implant-borne prosthesis. The subperiosteal implant should be considered as a long-term treatment option in those patients who are unable to wear a lower full denture due to resorption of the ridge and osseous defects that provide inadequate bone for placement of endosseous blade- or root-form implants. This case demonstrates the long-term viability of the subperiosteal implant. PMID- 7473870 TI - The press-fit/tap-in, anti-rotational vent-plant cylinder implant. AB - An unique press fit/tap-in, anti-rotational cylinder implant is described. It has an apical core and side vents which encourage bone growth, and its tapered fins provide a self-tapping mechanism, thus creating a firm anti-rotational mechanical anchorage from the moment of its insertion. Preliminary clinical trials have proved to be very successful, especially in poor bone density areas. PMID- 7473871 TI - Implant isotopy (II). AB - Because nature has given humans from 14 to 16 teeth per arch, the author maintains that an implantologist ought to replace each missing tooth with an implant and calls this philosophical creed "implant isotopy". Three different prosthetic solutions are suggested for the cases of either complete or partially edentulous arches, where from 12 to 14 abutments are emerging from the gingiva. The first type consists of parallel protruding posts. The second type is a modification of the first method through the connection of all the posts (although they are not perfectly parallel) via a titanium wire-drawn bar to be welded via endoral welding. Finally, the third type consists of a laboratory cast titanium denture composed of different sections that can be connected to one another and to the implant abutments via endoral welding. PMID- 7473872 TI - Oral-burn syndrome. AB - A possible major etiology is presented for previously unexplained throat and oral ulceration, inflammation and scarring, and also tissue loss associated with dental implant fixtures, oral surgery and periodontal flaps, and diffuse facial and pulpal pain. A simple solution is suggested. PMID- 7473873 TI - Prospective investigation of the single-crystal sapphire endosteal dental implant in humans: ten-year results. AB - This study conducted a longitudinal prospective clinical study of the single crystal sapphire (Al2O3) cylindrical screw-shaped endosteal dental implant, and attempted to establish clinical parameters to evaluate implant success or failure. Twenty-eight mandibular implants (17 patients) were placed. After six weeks' healing, 23 implants in 15 patients served as distal abutments for fixed prostheses (baseline). Implants were evaluated for bleeding index, crevicular fluid volume index, plaque accumulation index, radiographic index, mobility index, and patient comfort. Any implant failing in three of these criteria or implants removed were judged as failures. After 10 years, of the 21 baseline implants recalled (two implants were lost to recall), 17 were fully functional, for an 81% success rate. The use of qualitative and quantitative clinical evaluation parameters as utilized in this study appears to be important and useful in assessments of the clinical serviceability of dental implants. These parameters can be used in human clinical trials as well as in experimental animal studies. PMID- 7473874 TI - Generation of cell lines from embryonic quail retina capable of mature neuronal differentiation. AB - The avian embryonic retina is widely used as a model system for cellular and molecular studies on central nervous system neurons. We aimed at the generation of cell lines from the early embryonic quail retina by retroviral oncogene transduction. For this, we made use of the retina organ culture system which exhibits both proliferation, necessary for stable oncogene transduction, and initial neuronal differentiation, a prerequisite for the generation of cell lines with mature neuronal properties. The oncogene myc was chosen as it is both proliferation-inducing and differentiation-compatible. A chimeric gene, mycER, containing v-myc and the hormone-binding domain of the estrogen receptor, was used for transduction in order to allow for hormone regulation of myc activity. Transduced organ-cultured cells from temporal and nasal retina were passaged into sparse single cell cultures. From these, colonies of rapidly dividing cells were isolated and the progeny expanded as cell lines. The lines contained cells with features of neuroepithelial cells, showing vimentin and A2B5. They also contained spontaneously differentiated neuronal cells showing neurofilament L and N-CAM180. A subpopulation of the neuronal cells exhibited the morphological characteristics of retinal ganglion cells, i.e., large pear-shaped somata each emitting one long process with a distinct growth cone. In addition, they showed the marker profile of retinal ganglion cells, i.e., expression of Thy-1, G4, DM-GRASP, Nr-CAM, neurofilament H, and tau. Neuronal differentiation could be induced by the addition of db cAMP and retinoic acid. The mature neuronal features of the lines open new possibilities to study properties of retinal neurons, including ganglion cells, in a defined and manipulable experimental system. PMID- 7473875 TI - Protein kinase C modulates exogenous acetylcholine current in Xenopus oocytes. AB - The modulation of acetylcholine-activated current (IACh) by protein kinase C (PKC) was studied in Xenopus laevis oocytes microinjected with either mRNA extracted from C2C12 myotubes (C2C12 mRNA) or RNAs encoding murine alpha beta gamma delta subunits of the nicotinic ACh receptor (nAChR). Voltage-clamped oocytes were treated for 90 sec with 12-O-tetradecanoylphorbol-13-acetate (TPA, 300 nM), a potent PKC activator. Transient increase in the amplitude and acceleration in the decay of IACh were invariably observed within minutes of TPA application, and were independent of extracellular Ca2+ concentration. Both parameters recovered to control within 20-30 min; then a slight depression of IACh developed. By this time, an initial PKC down regulation was observed. At the peak of TPA-induced potentiation, dose-response relations suggested an increased binding affinity of nAChR for the neurotransmitter. 4 alpha-phorbol 12,13 didecanoate (300 nM), a biologically inactive analogue of TPA, did not affect IACh, while staurosporine (5-10 microM), a potent inhibitor of PKC activity, suppressed the action of TPA on IACh. In oocytes co-injected with C2C12 mRNA and with rat brain mRNA, IACh was potentiated by 5-hydroxy-tryptamine (10 microM), whose receptors are coupled to phosphoinositide hydrolysis. The nAChR-channel activity in cell-attached patches increased when TPA was applied to the oocytes. In 50% of the oocytes examined, a sustained depression of the single channel activity followed. We conclude that in Xenopus oocytes an endogenous PKC system regulates the function of embryonic-type muscle nAChRs. PMID- 7473876 TI - Glial fibrillary acidic protein mRNA isotypes: expression in vitro and in vivo. AB - Glial fibrillary acidic protein (GFAP) and its mRNA, primarily expressed in astrocytes, are also expressed in peripheral nervous system Schwann cells as well as in certain non-neural tissues. Schwann cells express a GFAP mRNA (GFAP-beta) which differs from the CNS-type mRNA (GFAP-alpha) by the presence of an extended 5' untranslated region. We have developed a polymerase chain reaction assay which allows distinction of these two GFAP mRNAs, as well as quantitative analysis of their levels. In the cultured rat Schwannoma cell line RT4-D6, GFAP-beta was the major GFAP mRNA species, accounting for at least 75% of total GFAP (alpha + beta) mRNA. GFAP-beta was also detected in primary rat astrocyte cultures, where it constituted approximately 5% of the total GFAP mRNA, as well as in RNA samples prepared from normal rat cerebral cortex, and from hamster and human brain. In rat cortex, the temporal expression of GFAP-beta mRNA paralleled that of total GFAP mRNA, with plateau levels reached between postnatal days 15 and 20. In astrocyte cultures, the relative levels of GFAP-alpha and -beta mRNAs were differentially regulated by exposure to interferon-gamma (10 to 25 units/ml), which caused an increase in GFAP-beta levels while at the same time no change or a small decrease in total GFAP levels. In rat brain cortical slices, 4 hr exposure to 25 units/ml interferon-gamma decreased total GFAP mRNA levels over tenfold, while GFAP-beta levels were unaffected. These data indicate that a second form of the GFAP mRNA is expressed in astrocytes both in vivo and in vitro and provide evidence for independent regulation of these two GFAP mRNA species. PMID- 7473877 TI - Expression of the nerve growth factor gene is controlled by the microtubule network. AB - Colchicine, nocodazol, and vinblastine, three microtubule-disrupting drugs, were shown to increase the levels of both nerve growth factor (NGF) mRNA and cell secreted NGF protein in L929 cells, with levels of glyceraldehyde-3-phosphate dehydrogenase (GAPDH) or amyloid precursor protein (APP) mRNAs remaining unaffected. Northern blot analysis demonstrated that colchicine also increased NGF mRNA levels in rat primary astrocytes and mouse skin fibroblasts. The specificity of the effects observed was assessed by the fact that the microtubule stabilizing agent Taxotere, a semisynthetic compound structurally related to taxol, suppressed the effects of colchicine, whereas lumicolchicine, a colchicine derivative that has no action on the microtubule network, had no influence on NGF expression. Likewise, the disruption of the microfilament network by cytochalasin B did not increase NGF mRNA levels in L929 cells. Furthermore, the increase in NGF gene expression observed following microtubule disruption depended on a cascade of events involving at least one protein kinase, which is not down regulated by phorbol ester, and on a pertussis toxin sensitive step. These results support the concept that tubulin and/or the microtubule cytoskeleton play an active role in the regulation of the NGF gene. PMID- 7473878 TI - Alteration of corticotropin-releasing factor immunoreactivity in MPTP-treated rats. AB - A decrease of corticotropin-releasing factor (CRF) concentration has been reported in patients with Parkinson's disease (PD). The present study further examined the role of CRF in an animal model of parkinsonism induced by 1-methyl-4 phenyl-1,2,3,6-tetrahydropyridine (MPTP). Results indicated that both subchronic (2 days) and chronic (7 days) MPTP treatments decreased the number of CRF immunoreactive neurons in both the paraventricular nucleus (PVN) of the hypothalamus and the central nuelcus of the amygdala (ACN). This effect lasted for almost a month after withdrawal of chronic MPTP injections. In addition, nomifensine pretreatment protected against MPTP's toxicity on DA neurons, as assessed by tyrosine hydroxylase immunoreactivity in the substantia nigra. However, the same treatment did not prevent the toxicity of MPTP on CRF neurons. Further, no significant difference was notable in the number of CRF immunoreactive neurons between normal young adult and normal middle-aged rats in both the PVN and the ACN. These results suggest that MPTP also produces a neurotoxicity on CRF neurons, and this effect is not secondary to MPTP's effect on DA neurons. Besides, altered CRF neuronal activity is involved in the process of pathological ageing, but not physiological ageing. Further, reduced CRF immunoreactivity in the PVN and ACN may imply alterations of neuroendocrine, autonomic as well as central functions caused by MPTP. PMID- 7473879 TI - Collagen implants and cortico-spinal axonal growth after mid-thoracic spinal cord lesion in the adult rat. AB - We describe an experimental model to study regeneration of lesioned corticospinal tract (CST) fibers in the adult rat spinal cord. After transection of all CST fibers at mid-thoracic level the gap is grafted with a sterile, cell-free collagen matrix. Two methods of collagen-application are used: 1) injection of a fluid collagen solution into the lesioned area which self-assembles in situ and 2) implantation of a solid collagen gel. At 4 weeks post-implantation CST axons are anterogradely labelled with horseradish-peroxidase (HRP). The collagen implant is evaluated for ingrowth of CST axons. The histopathological reaction (gliotic response) around the lesion and within the matrix is also studied. After application of a fluid collagen solution into the lesion area HRP-labelled CST axons can be visualized within the implant. In addition, astroglial and reactive microglial cells invade the collagen-matrix. On the other hand, if collagen is implanted as an already self-assembled gel, no ingrowth of labelled CST axons nor of astroglial/reactive microglial cells is observed. Both methods of collagen application result in a considerable reduction of the gliotic response as compared to the ungrafted animals. We conclude that the method of application of collagen (i.e., fluid or gel) considerably affects the response of lesioned CST axons. The application of a fluid collagen graft which in situ self-assembles is beneficial for the regrowth of lesioned CST axons in rat spinal cord. In this respect the formation of an astroglial scaffolding structure within the (fluid) collagen, probably due to optimal integration between host and graft, is very important. The inability of injured CST fibers to enter the solid collagen graft may be related to the absence of an astroglial scaffolding structure within the implant. PMID- 7473880 TI - Gangliosides have a bimodal effect on DNA synthesis in U-1242 MG human glioma cells. AB - GM1, GD1a, and GT1b inhibit both PDGF-stimulated and serum-stimulated DNA synthesis in Swiss 3T3 cells and the human glioma cell line U-1242 MG in a dose dependent manner. The ganglioside inhibitory effect is counteracted in a dose responsive fashion by serum such that ganglioside-induced inhibition is essentially abolished in 10% serum. Because of the potentially important role that gangliosides play in growth regulation of human gliomas, this phenomenon was studied in detail using U-1242 MG cells. Stimulation of DNA synthesis by low doses of serum in U-1242 MG cells is inhibited in a dose-responsive fashion by ganglioside GM1. However, serum itself counteracts the inhibitory effect of ganglioside in a dose responsive way. Kinetic analyses demonstrate that GM1 competes with some components of serum for sites on U-1242 MG cells (Kb of GM1 = 12.5 microM). On the other hand, GM1, GD1a, and GT1b stimulate DNA synthesis in quiescent U-1242 MG cells in both sparse and confluent conditions, indicating that ganglioside-stimulated DNA synthesis is dependent on the phase of cellular growth rather than cellular density. This growth stimulatory effect of gangliosides is more potent on quiescent, confluent cells than quiescent, sparse cells. These results demonstrate that exogenously added gangliosides can have opposite (bimodal) effects on progression of human glioma cells through the cell cycle depending upon the growth phase of the cells. PMID- 7473881 TI - Laminin oligopeptide derivatized agarose gels allow three-dimensional neurite extension in vitro. AB - The phenotypic expression of various neural cells is influenced by extracellular matrix (ECM) molecules. This study aims to develop a three-dimensional gel tailored to support neurite extension from neural cells. Laminin-derived (LN) oligopeptides CDP-GYIGSR, a 19-mer IKVAV containing sequence, GRGDSP, a cocktail of the three aforementioned LN peptides (PEPMIX), and a control peptide sequence GGGGG were covalently linked to an agarose hydrogel backbone using the bi functional coupling agent 1'1, carbonyldiimidazole. Embryonic day 9 chick DRGs and PC12 cells were suspended in three dimensions in underivatized and derivatized agarose gels and neurite extension was analyzed. Agarose gels derivatized with CDPGYIGSR and PEPMIX enhanced neurite outgrowth from DRGs while GRGDSP and IKVAV derivatized gels inhibited neurite extension when compared to underivatized agarose gels. The IKVAV derivatized gels significantly enhanced neurite outgrowth from PC12 cells in comparison to underivatized and other LN peptide derivatized gels. Agarose hydrogels carrying covalently immobilized LN oligopeptides thus evoke specific responses from cells which contain receptors to the peptides used. Agarose hydrogels derivatized with neurite promoting peptide sequences may find applications in various models of in vivo regeneration of nervous tissue. PMID- 7473882 TI - Membrane properties of cultured glial cells obtained mechanically or by enzymatic dissociation: differential alterations by gangliosides. AB - Rat astroglial cells were obtained either mechanically or by enzymatic dissociation and some properties of the primary and secondary cultures were studied. Differences in the ganglioside amount, taurine uptake, membrane fluorescence anisotropy, or their respective modulation by total brain gangliosides confirmed that although the two types of cultures are morphologically similar, they are biochemically distinct. PMID- 7473883 TI - Signal transduction alterations in peripheral nerves from streptozotocin-induced diabetic rats. AB - We have previously determined the presence of muscarinic receptors and the expression of several G proteins in homogenates and myelin fractions from rat sciatic nerves. In the present study we investigated whether changes in several signal transduction pathways in peripheral nerves might be responsible for some of the biochemical abnormalities (e.g., phosphoinositide metabolism) present in sciatic nerves from streptozotocin-induced diabetic rats. Sciatic nerves from 5 week diabetic rats that were prelabelled with [3H]-myo-inositol displayed a significant increase in the basal release of inositol mono- and bis-phosphate, while carbamylcholine-stimulated release was significantly smaller. Basal- and forskolin-stimulated adenylyl cyclase activity was significantly decreased in sciatic nerve homogenates from diabetic animals. However, we were unable to detect any significant differences in the levels of cAMP in intact nerves or in nerve segments that were incubated in the presence or absence of forskolin. ADP ribosylation experiments showed that in sciatic nerves from experimentally diabetic rats there was a significant increase in the ADP-ribosylation catalyzed by cholera and pertussis toxins. Measurements of the levels of alpha-subunits of G proteins revealed that the expression of Gq/11 alpha, Gs alpha, and Gi-3 alpha was increased by 30 to 50%. These results indicate that during the course of experimental diabetes, peripheral nerves exhibit an abnormal production of inositol phosphates and cAMP, together with an abnormal expression and/or function of G proteins. One of the consequences of such alterations is the diminished release of inositol phosphates triggered by muscarinic agonists in diabetic sciatic nerves. PMID- 7473884 TI - Syngeneic bone marrow transplantation eliminates V beta 8.2 T lymphocytes from the spinal cord of Lewis rats with experimental allergic encephalomyelitis. AB - Experimental allergic encephalomyelitis (EAE), an animal model for multiple sclerosis (MS), is a paralytic disease of the central nervous system (CNS) mediated by T-lymphocytes reactive to myelin basic protein (MBP). Lewis rats actively immunized with fragment 68 to 82 of guinea pig MBP develop a monophasic disease with spontaneous recovery. Lymphocyte recognition of the primary encephalitogenic sequence of MBP (fragment 68 to 82) is V beta 8.2 T cell receptor (TCR) skewed [1-3]. Lewis rats in clinical remission at 1 month and 3 months after spontaneous resolution of EAE retain V beta 8.2 T-lymphocytes in the CNS when analyzed by reverse transcriptase polymerase chain reaction or in situ hybridization. In contrast, 1 and 3 months after clinical remission from syngeneic bone marrow transplantation, V beta 8.2 T lymphocytes are absent from the CNS. During clinically active EAE and inflammatory breakdown of the blood brain barrier, immune ablation and reconstitution with syngeneic bone marrow results in clinical tolerance of the new immune system to myelin. PMID- 7473885 TI - Spatial distribution of omega-agatoxin IVA binding sites in mouse brain slices. AB - A peptide toxin derived from funnel-web spider venom, omega-agatoxin IVA, blocks voltage-sensitive calcium channels. Many pharmacological and electrophysiological studies have shown that these channels are widely distributed in both the central nervous system (CNS) and neuromuscular junctions. However, a direct morphological demonstration of the binding sites of this toxin is still lacking. To identify which cells have the binding sites, a biologically active, biotin-conjugated omega-agatoxin IVA was applied to mouse cerebellar and hippocampal slices. Confocal microscopy revealed that omega-agatoxin IVA binding sites were distributed on the somata of Purkinje cells, cerebellar granule cells and interneurons, as well as on the dendrites of Purkinje cells. In the hippocampus, the binding sites were localized on the somata of pyramidal cells of the CA1-CA4 region and on the somata of granule cells in the dentate gyrus. A sequential competitive reaction confirmed the specificity of the binding in the cerebellum and CA1 pyramidal cells, and also suggested a difference in the binding affinity between CA1 and CA3 pyramidal cells. Since a high concentration of omega-agatoxin IVA (2 microM) was needed for the present study, the omega-agatoxin IVA binding sites presented in this study may represent "P-type" and "Q-type" calcium channels. PMID- 7473886 TI - Platelet-derived growth factor is a survival factor for PSA-NCAM+ oligodendrocyte pre-progenitor cells. AB - Mature oligodendroglia, which synthesize and express lipids and proteins characteristic of myelin, are generated from precursor cells which are formed in germinal matrix, then migrate widely through the neuraxis. We now demonstrate that these precursor cells can be recognized at a very early stage by their surface expression of polysialylated neural cell adhesion molecules (PSA-NCAM), and only later bind anti-ganglioside antibodies that had previously been used to recognize "O-2A" oligodendroglial precursor cells. PSA-NCAM expression by these cells is likely to be of functional significance, since a recent study demonstrated that O-2A cells become immobile when stripped of PSA-NCAM. Platelet derived growth factor (PDGF) proved to be a survival factor for these PSA NCAM+cells, and in a defined medium, PDGF was sufficient to ensure maturation of immunopurified PSA-NCAM+cells to oligodendroglia. PMID- 7473887 TI - Notochord and floor plate stimulate oligodendrocyte differentiation in cultures of the chick dorsal neural tube. AB - The regionalization of oligodendrocyte potentialities and the cellular interactions leading to the expression of the oligodendrocyte phenotype have been analyzed in the embryonic chick spinal cord. Dorsal and ventral regions of the spinal cord of 4-day-old embryos (E4) were cultivated separately. Oligodendrocyte differentiation was monitored at various times after explantation, using specific oligodendrocyte markers. After 2 weeks, several hundreds of differentiated oligodendrocytes were invariably observed in ventral cultures whereas significant numbers of oligodendrocytes failed to develop in dorsal spinal cord cultures. However, the E7 dorsal spinal cord was found to produce large numbers of oligodendrocytes, indicating that the ventral restriction of oligodendrocyte potentialities is transient. To test whether ventrally derived signals might influence oligodendrocyte differentiation, E4 dorsal spinal cord microexplants were cocultivated with notochord segments or with floor plate tissue. Numerous oligodendrocytes were found in dorsal explants associated with either tissue, notochord or floor plate, but not in dorsal explants cultivated alone, indicating that cells competent to be induced along the oligodendrocyte phenotype exist in the dorsal spinal cord. These results show that oligodendrocyte differentiation potentialities are initially restricted to the ventral spinal cord and suggest that ventrally derived signals from notochord and floor plate influence oligodendrocyte differentiation in the embryonic spinal cord. PMID- 7473888 TI - beta-Amyloid peptide fragment 25-35 potentiates the calcium-dependent release of excitatory amino acids from depolarized hippocampal slices. AB - beta-Amyloid protein (beta AP) has been frequently associated with the neuropathology of Alzheimer's disease (AD), although the mechanisms by which it can induce neurodegeneration are still unknown. Some studies in hippocampal cultured neurons suggest that beta AP, particularly its fragment 25-35, may induce neural growth or render neurons more vulnerable to excitotoxic insults by a mechanism involving intracellular Ca2+ dyshomeostasis. We have studied the effect of fragment 25-35 on the release of endogenous amino acids from hippocampal slices of young adult (3-3.5-month-old) and aged (23-25-month-old) rats, under basal, K(+)-depolarization, and post-depolarization conditions, in the presence and absence of Ca2+. In both young and aged tissue, the basal release of amino acids was not affected by the peptide. By contrast, 1-hr preincubation of slices from young animals with 10 microM 25-35 fragment resulted in a 140% increase of glutamate and aspartate release stimulated by K+ depolarization, compared with the control-stimulated release. These effects were strictly dependent on external Ca2+. Neither the K(+)-stimulated release of gamma amino butyric acid (GABA) nor the release of glycine, glutamine, taurine, or alanine, which was not stimulated by high K+, were affected. Substance P and a scrambled sequence of the 25-35 fragment were without any effect per se, but substance P blocked the stimulatory effect of fragment 25-35 on glutamate and aspartate release. In slices from aged rats the basal release of glutamate was significantly higher (260%) than that in young tissue, and the K(+)-induced release of both aspartate and glutamate was also higher.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7473889 TI - Concentrations of metal elements in mouse organs after intratracheal administration of coal fly ash. AB - Size-density fractionated coal fly ash particles, 1-5 microns in diameter and 2.4 2.8 g/cm3 in density, were administered to mouse lungs intratracheally. The change in the concentrations of metal elements in the lungs, kidneys, livers and spleens was measured by inductively coupled plasma atomic emission spectroscopy and atomic absorption spectroscopy up to 16 weeks after administration. In the lungs, each elemental concentration increased 1 week after administration and decreased thereafter with time after administration. For example, the concentrations of Al were 1018 +/- 272 micrograms/g tissue at 1 week and decreased thereafter to 556 +/- 177 micrograms/g tissue at 16 weeks after administration. The concentrations of Ca, Mn and Ni decreased more rapidly than the other elements, suggesting that Ca, Mn and Ni were dissociated from the alumino-silicate matrix of the fly ash particles in the lungs. In the kidneys, the concentration of Ni significantly increased at 2, 8 and 16 weeks after administration; the concentration of Ni showed an increase of 0.4 +/- 0.2 micrograms/g over the control mouse at 16 weeks after administration. In the livers, increased concentrations of Ca and Mn were observed at 2, 4 and 16 weeks. On the other hand, no changes in the concentrations of these elements were observed in the spleen. PMID- 7473891 TI - Effect of alkaline ionized water on reproduction in gestational and lactational rats. AB - Alkaline ionized water (AKW) produced by electrolysis was given to gestational and lactational rats, and its effect on dams, growth of fetuses and offsprings were investigated. The results showed that the intake of food and water in dams increased significantly when AKW was given from the latter half of the gestation period and from the former half of the lactation period. Body weight of the offsprings in the test group, both males and females, increased significantly from the latter half of the lactation period. During the lactation period and after weaning, the offsprings in the test group showed significantly hastened appearance of abdominal hair, eruption of upper incisors, opening of eyelids and other postnatal morphological developments both in males and females, as well as earlier separation of auricle and descent of testes in males compared with the control was noted. As mentioned above, it was suggested from the observations conducted that the AKW has substantial biological effects on postnatal growth, since intake of food and water and body weight of the offsprings increased and postnatal morphological development was also accelerated. PMID- 7473890 TI - Reproductive and developmental toxicity studies of toluene. I. Teratogenicity study of inhalation exposure in pregnant rats. AB - Toluene is a widely used solvent in industry which is the subject of abuse among the younger generation. A teratogenicity study of toluene by inhalation exposure was carried out in Sprague-Dawley rats and the effects on dams, fetuses and offspring were assessed. Pregnant females were exposed to 600 or 2000 ppm toluene for 6 h/day from day 7 to day 17 of pregnancy. The control group inhaled conditioned clean air under the same exposure conditions. Maternal exposure to 2000 ppm toluene caused significant toxic effects such as body weight suppression of dams and offspring, high fetal mortality and embryonic growth retardation, but no external, internal or skeletal anomalies were observed in the fetuses of any treated group. In addition, there were no differences in the results of pre- and postweaning behavioral tests of the offspring. However, no toxic or teratogenic changes which could be related to toluene exposure were apparent in the 600 ppm group. Further studies are warranted with toluene at higher concentrations applied during the period of organogenesis. PMID- 7473892 TI - Antidotal action of atropine sulfate against insecticide benfuracarb poisoning in rats. AB - The antidotal action of atropine sulfate and 2-pyridine aldoxime methiodide (2 PAM) against poisoning attributable to the new procarbamate insecticide benfuracarb [(ethyl N-[2,3-dihydro-2,2-dimethylbenzofuran-7-yloxycarbonyl (methyl) aminothio]-N-isopropyl-beta-alaninate] was compared utilizing rats as our experimental model. Both the intraperitoneal and oral administrations of these antidotes were examined after five, ten, fifteen and thirty minutes exposure periods, following treatment with benfuracarb at dose levels approximating LD50 and LD100. The results obtained demonstrate that both the intraperitoneal and oral administrations of atropine sulfate blocked or significantly reduced the toxic effects of benfuracarb and protected the animals from death. The intraperitoneal administration route appears to be more effective than was the oral route. In addition, the administration of atropine sulfate after the shorter period (up to 15 minutes), following exposure to benfuracarb, improved antidotal action, particularly with the LD100 dose of benfuracarb. It is suggested that atropine sulfate antagonizes benfuracarb poisoning by blocking acetylcholine (ACh) receptors, as many other carbamate insecticides, since benfuracarb was an in vivo cholinesterase (ChE) inhibitor and the toxic effect of benfuracarb was reduced by atropine sulfate. 2-PAM, however, did not significantly block or reduce the toxic effects of benfuracarb. PMID- 7473893 TI - Radiosensitization by dimethylbenzanthracene, diphenylcyclopropenone and aminoanthraquinones. AB - Radiosensitization of mice by dimethylbenzanthracene, diphenylcyclopropenone and aminoanthraquinones was investigated in a model where survival time after lethal radiation was scored. Survival time was shortened by nontoxic doses of the chemicals. The used in vivo system confirmed the radiosensitizing potential of dimethylbenzanthracene reported previously with in vitro studies. Moreover, radiosensitizing properties of diphenylcyclopropenone and aminoanthraquinones could be demonstrated. The sensitizing interaction of these chemicals with radiation adds a new facet to their toxicological spectrum and could, by enhancing radiation effects, influence estimates of risk. On the other hand, diphenylcyclopropenone or aminoanthraquinones deserve consideration as topical sensitizers in conditions where radiation is indicated to treat cutaneous malignancies. PMID- 7473894 TI - Effects of medroxyprogesterone acetate and beta-estradiol on interleukin-6 production from osteoblasts and bone marrow macrophages of Wistar rats of different ages. AB - Using 18-week-old and 52-week-old Wistar rats, we examined interleukin-6 (IL-6) production from osteoblasts and bone marrow macrophages treated with medroxyprogesterone acetate (MPA) and/or beta-estradiol. The level of IL-6 production by osteoblasts was increased by treatment with MPA and, reversely, decreased by treatment with beta-estradiol. These changes were especially remarkable in osteoblasts of 52-week-old rats. Additionally, and in contrast, the production of IL-6 by bone marrow macrophages was not significantly changed by treatment with both agents. These data suggest that because the increased production of IL-6 by osteoblasts treated with MPA in opposition with beta estradiol, MPA should be careful for osteoporosis dependent upon osteoclasts activated by IL-6. Finally, there was a marked difference in the amount of IL-6 produced between osteoblasts and bone marrow macrophages. PMID- 7473895 TI - Depletion of glutathione and hepato-toxicity caused by vinyl ethers in mice. AB - 4-Nitrophenyl vinyl ether (NPVE) and phenyl vinyl ether (PVE) administered i.p. in mice lowered hepatic non-protein sulfhydryl (NP-SH) content, but did not elevate the serum glutamate pyruvate transaminase (GPT) activity. n-Butyl vinyl ether (BVE) showed no significant effects either on the NP-SH content or on the serum GP activity. Mice pretreated with buthionine sulfoximine were sensitive to the potential toxicity of NPVE. These results showed that aryl vinyl ethers, NPVE and PVE, are more toxic than the alkyl vinyl ether, BVE, and that glutathione plays an important role on the protection of hepatic injury by reactive metabolite(s) derived from vinyl ethers. PMID- 7473896 TI - Thoughts suggested by a recent paper: questions on non-parametric analysis of quantitative data. PMID- 7473897 TI - Predictability of clinical adverse reactions of drugs by general pharmacology studies. PMID- 7473898 TI - Effects of the neonatal vitamin A exposure on behaviors of adult rats. AB - The effects of neonatal vitamin A exposure on six behavioral parameters in adult Wistar rats were investigated. Newborn male rats were injected intraperitoneally with 80,000 IU/kg/day of retinol palmitate (vitamin A) dissolved in physiological saline or vehicle alone (controls) during 1-5 days after birth. The animals were examined using the open field test (at 36 weeks of age), the rotarod test (38 weeks), the Biel water-maze test (40 weeks), the conditioned avoidance test (42 weeks), the running-wheel activity with pentobarbital challenge test (44 weeks), and the tail flick test (45 weeks). The vitamin A-exposed rats made significantly more errors and took significantly longer times in the Biel water-maze test, and showed significantly slower responses in the tail flick test than the controls. No significant effects of treatment with vitamin A were observed in any of the other behavioral tests. These results indicate that a long-lasting defect in learning in water-maze task and depression of the heat-pain response are induced in rats exposed neonatally to vitamin A. PMID- 7473899 TI - Salt and solution: physician heal thyself. PMID- 7473900 TI - Reprogrammed macrophage tumor necrosis factor and interleukin-1 release with inflammatory pretreatment: differential regulation by endotoxin and zymosan. AB - OBJECTIVE: To determine whether different reprogrammed alterations in endotoxin (lipopolysaccharide, LPS)-stimulated tumor necrosis factor (TNF) and interleukin 1 (IL-1) release are seen following pretreatment with endotoxin (LPSp) or pretreatment with the particulate inflammatory stimulus zymosan. METHODS: Murine peritoneal macrophages (M phi) pretreated for 24 hours in vitro with medium, LPSp, zymosan, latex beads, or killed Escherichia coli. After 24 hours M phi were restimulated with medium, LPSa, zymosan, latex beads, or E. coli, M phi supernatant TNF and IL-1 were measured after 24 hours and mRNA levels determined after 6 hours with reverse-transcriptase polymerase chain reaction. RESULTS: Pretreatment with low dose LPSp markedly inhibited TNF release by both LPSa or zymosan, while pretreatment with zymosan increased LPSa-stimulated TNF release. Pretreatment with both LPSp and zymosan augmented LPSa and zymosan-stimulated IL 1. Zymosan pretreatment augmentation of TNF and IL-1 was accompanied by lower than basal levels of cytokine message. CONCLUSION: Reprogrammed macrophage TNF and IL-1 release was differentially regulated by distinct inflammatory stimuli. Understanding reprogrammed macrophage cytokine regulation may enable specific therapy to modify dysregulated cytokine release during sepsis and trauma. PMID- 7473901 TI - Early neutrophil sequestration after injury: a pathogenic mechanism for multiple organ failure. AB - Polymorphonuclear neutrophils (PMNs) play a pivotal role in the inflammation that precedes multiple organ failure (MOF). In a rat model of MOF, PMNs become primed for enhanced superoxide anion (O2-) release and CD11b expression, sequester in end organs, and produce organ failure. Therefore, we hypothesized that circulating PMNs harvested in the first 24 hours after injury from trauma patients at risk for MOF would (1) exhibit a primed O2- release, (2) upregulate CD11b expression, and (3) show evidence of sequestration in tissues. Extracellular PMN O2- release and CD11b receptor expression were measured at 3, 6, 12, and 24 hours after injury in 33 torso trauma patients with Injury Severity Scores > 15; eight patients (24%) developed MOF. Healthy adults served as controls. PMNs after injury were primed for enhanced in vitro O2- release at 3, 6, 12, and 24 hours after injury, indicating prior in vivo priming. CD11b expression was also increased at 6, 12, and 24 hours after injury. Circulating PMN numbers increased sharply at 3 hours after injury, before decreasing dramatically at 6 and 12 hours, suggesting end organ sequestration. At 12 hours after injury, declines in circulating PMNs were significantly greater in MOF than in non-MOF patients (p < 0.05). These data indicate that PMNs are quickly mobilized into the circulation after injury and then primed for enhanced O2- release and CD11b expression. PMN priming appears to be a necessary preamble to PMN sequestration in patients with major torso trauma. Upregulation of PMN function, accompanied by subsequent end organ sequestration, may represent an important early event in the pathogenesis of MOF after injury. PMID- 7473902 TI - Inhaled nitric oxide variably improves oxygenation and pulmonary hypertension in patients with acute respiratory distress syndrome. AB - OBJECTIVE: The purpose of this study was to determine the effect of inhaled nitric oxide (NO) on oxygenation, hemodynamics, and ventilation in patients with adult respiratory distress syndrome (ARDS). METHODS: Sixteen trials of inhaled NO were performed in 14 ARDS patients (age 42.9 +/- 6) who had significant pulmonary hypertension (mean pulmonary artery pressure > 30 mm Hg). Patients were monitored with arterial and pulmonary artery catheters. Volume ventilation was performed by a strict protocol. Data were collected at baseline and 30 minutes after 20 and 40 ppm NO. RESULTS: Overall, the Pao2/FIO2 ratio increased from 69.5 +/- 3.9 to 100.8 +/- 9.5 at 20 ppm, a 42.9 +/- 8.7% increase, and to 97.7 +/- 13.1 at 40 ppm, a 44.1 +/- 14.2% increase over baseline (p = 0.001). However, 5/16 trials demonstrated minimal improvement (Pao2/FIO2 ratio increase < 20%), 5 moderate improvement (20-50%), and 6 marked improvement (> 50%). Overall the mean pulmonary artery pressure (MPAP) decreased from 41.1 +/- 1.8 mm Hg to 34.3 +/- 1.3 at 20 ppm and 33.5 +/- 1.8 mm Hg at 40 ppm, a 15.0 +/- 3.7% decrease, p = 0.002. However, 5/16 trials demonstrated minimal improvement (MPAP decrease < 10%), 7 moderate improvement (10-20%), and 4 marked improvement (> 20%). At 40 ppm NO, 4 patients with moderate improvement in MPAP at 20 ppm NO had marked improvement. There were no changes in systemic oxygen delivery and consumption, intrapulmonary shunt, or lung compliance at 20 or 40 ppm (p > 0.05). Following collection of these data, prolonged NO inhalation (2-20 ppm) was done in 10 of 14 patients for 4.5 +/- 0.7 days (range 1-10 days). Overall, there were 7 deaths (mortality 50%). In patients receiving prolonged NO, there were 3 deaths (mortality 30%). All 4 patients who failed to respond to inhaled NO subsequently died (mortality 100%). CONCLUSIONS: Inhaled NO improves oxygenation and pulmonary hemodynamics in patients with ARDS. However, the improvement to NO is variable. PMID- 7473904 TI - Clearing the cervical spine in obtunded patients: the use of dynamic fluoroscopy. AB - OBJECTIVE: Obtunded patients (Glasgow Coma Scale score of < 13) with normal cervical roentgenograms remain in collars until they can be clinically evaluated. Cervical collars provide incomplete immobilization and have complications. Our hypothesis was that cervical spines could be evaluated in obtunded patients with normal cervical roentgenograms using dynamic fluoroscopy. DESIGN: This study was a prospective clinical evaluation. METHODS: Obtunded trauma patients with normal cervical roentgenograms underwent fluoroscopic examination of the cervical spine through a full range of motion. RESULTS: Fluoroscopic evaluations were done in 116 patients. There were 113 true negative examinations. Two patients had facet fractures not diagnosed on cervical roentgenograms, and no instability on fluoroscopy. One patient had a positive exam, with 2 mm of subluxation. There were no neurologic complications. Decubiti were present in 44% and were more frequent when the collar was on > 5 days (p = 0.029). CONCLUSIONS: Dynamic fluoroscopy can safely and effectively clear the cervical spine in obtunded patients. Earlier removal of the collar decreases decubiti. PMID- 7473903 TI - Portal triad injuries. AB - OBJECTIVE: Injuries to the portal triad are a rare and complex challenge in trauma surgery. The purpose of this review is to better characterize the incidence, lethality, and successful management schemes used to treat these injuries. DESIGN: A retrospective review of the experience of eight academic level I trauma centers over a combined 62 years. RESULTS: A retrospective review of the experience of eight anatomical structures of the portal hepatis: 118 injuries to the anatomical structures of the portal hepatis: 55 extrahepatic portal vein injuries, 28 extrahepatic arterial injuries, and 35 injuries to the extrahepatic biliary tree. Sixty-nine percent of the injuries were by penetrating mechanism and 31% were by blunt mechanism. All patients had associated injuries with a mean Injury Severity Score of 34 in blunt trauma patients. Overall mortality was 51%, rising to 80% in patients with combination injuries. Sixty-six percent of deaths occurred in the operating room, primarily from exsanguination; 18% of deaths occurred within 48 hours of injury from refractory shock, coagulopathy, or cardiac arrest; 16% occurred late. Ten percent of patients undergoing portal vein ligation survived, compared to 58% managed by primary repair. Survival after hepatic artery ligation was 42%, compared to 14% after primary repair. Survival after biliary-enteric anastomosis as treatment of extrahepatic bile duct injury was 89%, compared to 50% after primary repair and 100% after ligation of lobar bile duct injuries. Missed bile duct injuries had a high (75%) severe complication rate. CONCLUSIONS: Injuries to the anatomical structures of the portal triad are rare and often lethal. Intraoperative exsanguination is the primary cause of death, and hemorrhage control should be the first priority. Bile duct injuries should be identified by intraoperative cholangiography and repaired primarily or by enteric anastomosis; lobar bile ducts can be managed by ligation. PMID- 7473905 TI - Open fractures of the patella: long-term functional outcome. AB - Seventy-nine open patella fractures in 76 patients were treated between 1986 through 1994, with an 80% incidence of multiple injuries. All were treated with irrigation and debridement, open reduction, internal fixation, and reconstruction of the extensor mechanism. In no case was a primary patellectomy performed, even with severe comminution. There were three failures of initial fixation and one asymptomatic nonunion. Average range of motion for all groups was 112 degrees, at an average follow-up of 21 months. Secondary surgical procedures were performed in 65% of knees, the majority for symptomatic hardware. To determine long-term functional outcome, a modified Hospital for Special Surgery knee score was used. At an average of 36 months, good to excellent knee scores were observed in 17 of 22 patients. We conclude that all attempts for preservation of bone substance and precise reconstruction of the extensor should be attempted, reserving total patellectomy as a salvage procedure. PMID- 7473907 TI - Efficacy and safety of pneumococcal revaccination after splenectomy for trauma. AB - OBJECTIVE: To assess the outcome of patient education after splenectomy and vaccination and to determine the safety and efficacy of pneumococcal revaccination 2 or more years after primary vaccination. MAIN OUTCOME MEASURES: Titers to serotype no. 6 and no. 23 pneumococcus and cutaneous and systemic reaction to revaccination. RESULTS: A total of 112 consecutive postsplenectomy patients receiving pneumococcal vaccine were identified; 45 were contacted and offered revaccination; 24 patients demonstrated a lack of understanding of the postsplenectomy state (unaware of splenectomy n = 2, unaware of splenectomy risk n = 8, unaware of vaccine n = 23); 3 patients had infections requiring hospitalization (pneumonia, strep throat and tonsillitis, pneumonia and bacteremia); 40 patients agreed to revaccination, and 33 patients returned for follow-up titers; 16 of 33 (48%) demonstrated at least a two-fold increase in at least one titer. Only 15% described the revaccination as worse than a tetanus shot. CONCLUSIONS: (1) Despite physician-patient conversations, pamphlets, and Medic Alert bracelets, patient retention was poor. (2) All splenectomy patients should be revaccinated and reeducated between two and six years after splenectomy. (3) Revaccination after two years was well tolerated. (4) There were no fatal episodes of pneumococcal sepsis in over 200 patient years. PMID- 7473906 TI - Bioavailability of fluconazole in surgical intensive care unit patients: a study comparing routes of administration. AB - OBJECTIVE: Anticandidal therapy is commonly used in the surgical intensive care unit (SICU). Unfortunately, it is expensive because it is generally given intravenously, as acute trauma and abdominal surgery are often accompanied by impaired gastrointestinal function. We compared the systemic availability of fluconazole given enterally or intravenously in trauma and surgery SICU patients to determine the reliability of enteral administration. METHODS: Nine adult trauma (Injury Severity Score (> or = 18) and nine adult abdominal surgery SICU patients were randomized to receive fluconazole 100 mg via the intravenous (IV) or enteral route. Patients with a bilirubin > 4.0 mg/dL or creatinine clearance < 60 mL/min were excluded. Enteral fluconazole was crushed, dissolved, and flushed through a nasogastric or feeding tube. Eleven serial blood samples were drawn over 72 hours. Area under the curve (AUC), elimination rate constant (Ke), and terminal half-life (T1/2) were determined and compared (t test). Relative bioavailability was estimated (AUC(enteral)/AUCIV). RESULTS: Peak concentrations occurred within 2 hours after enteral dosing and 15 minutes after IV dosing. The relative bioavailability was 77%. Weight, AUC, Ke, and T1/2 did not differ between enteral and IV dosing. CONCLUSIONS: Fluconazole is significantly absorbed when crushed, dissolved, and given via a nasogastric or feeding tube in SICU patients. Nonsignificant trends toward lower systemic availability with enteral administration can be overcome with slightly higher doses. Since enteral administration of fluconazole costs 10% of fluconazole given intravenously, more liberal use of enteral administration offers tremendous savings. Such savings moderate the cost concerns of antifungal therapy. PMID- 7473908 TI - PASG: does it help in the management of traumatic shock? AB - A prospective, randomized study was designed to determine the efficacy of pneumatic antishock garment (PASG) in the treatment of traumatic shock in a medium-size urban community. A total of 291 traumatic shock patients were assigned to either the PASG or No-PASG treatment groups. Of these, data from 248 patients were analyzed in detail. Analysis of demographic factors--such as age, sex, and mechanism of injury--as well as prehospital evaluative tools--such as trauma and CRAMS scores, and injury severity scores--revealed that the two groups were well-matched. This study did not demonstrate significant differences in hospital stay or mortality between PASG and No-PASG patients. Similarly, in the subset of patients with blunt trauma, PASG was not found to be beneficial. PMID- 7473909 TI - Trauma care systems in urban Latin America: the priorities should be prehospital and emergency room management. AB - Trauma is a significant cause of premature death in developing nations, but financial resources to deal with it are extremely limited. To determine which segments of a developing nation's trauma system would be most amenable to improvements, we compared management and outcome of all seriously injured patients (Injury Severity Score of > or = 9 or died) treated over 1 year by the trauma systems associated with an urban hospital in Latin America, Regional Trauma Center 21 (n = 545) in Monterrey, Mexico, and a level I trauma center in the United States, Harborview Medical Center (n = 533) in Seattle, Wash. Mortality was higher in Monterrey (55%) than in Seattle (34%, p < 0.001), because of a preponderance of prehospital and emergency room (ER) deaths. In Monterrey, 40% of seriously injured patients died in the field and 11% in the ER, compared with 21% in the field and 6% in the ER in Seattle (p < 0.001). There were significant differences in prehospital care between the two trauma systems. Scene and transport times were < 30 minutes for 47% of Monterrey cases vs. 75% in Seattle (p < 0.001). For patients with arrival blood pressure < 80, prehospital intubations had been performed on 5% of Monterrey patients vs. 79% in Seattle (p < 0.001) and en route fluid resuscitation administered to 70% of Monterrey patients vs. 99% in Seattle (p < 0.001). The observed mortality patterns indicate that priorities for trauma system improvement in urban Latin America should focus on more rapid prehospital transport and improved en route and ER resuscitation.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7473910 TI - Cerebral blood flow and blood volume in response to O2 and CO2 changes in normal humans. AB - Changes in cerebral blood volume (CBV) after head injury may be an important determinant of intracranial pressure (ICP). To determine the normal response of CBV to hypoxemia, hypercapnia, and hypocapnia, eight normal subjects (5 males and 3 females; ages 25 to 43) were studied under these conditions. Cerebral blood volume was measured using an external collimated gamma detector to determine 99m Tc-labeled red blood cell (RBC) activity in the intracranial vascular pool, and cerebral blood flow (CBF) was determined by internal carotid artery duplex scanning. Hypocapnia (Paco2 = 26.0 +/- 1.7 mm Hg, mean +/- SE) was achieved by hyperventilation, hypercapnia (Paco2 = 47.8 +/- 1.5 mm Hg) was achieved by inhalation of 6% CO2, and hypoxemia (Pao2 = 38.1 +/- 1.1 mm Hg, O2 saturation = 76.7 +/- 2.0%) was achieved by inhalation of 10% O2. Changes in CBF and CBV were determined by comparing the values in each condition to the immediately preceding period of normoxia and normocapnia. For conditions of hypocapnia, hypercapnia, and hypoxemia, the percentage of change in CBV was: -7.2 +/- 0.01, 12.8 +/- 0.01, and 5.2 +/- 0.03, respectively. The simultaneous percentage of change in CBF for the same conditions was -30.7 +/- 4.0, 29.5 +/- 9.2, and 18.4 +/- 6.9, respectively. For all conditions, changes in CBF were greater than changes in CBV; however, this was most pronounced during hypocapnia induced by hyperventilation. Because the change in CBV reflects the potential change in ICP in response to treatment, therapeutic hyperventilation may impair CBF to a greater degree than it reduces ICP.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7473911 TI - U83836E reduces secondary brain injury in a rabbit model of cryogenic trauma. AB - Oxygen-derived free radicals have been implicated in secondary brain injury after head trauma. Although oxygen-free radicals may be derived from multiple sources, the activation and accumulation of neutrophils in areas of brain injury is a potential source of deleterious inflammatory mediators, including free radicals, which may exacerbate the primary insult. Neutrophils may also impede the cerebral microcirculation by congestion and occlusion of postcapillary venules. The lazaroid U83836E is a potent-free radical scavenger that inhibits lipid peroxidation. U83836E was studied in a cryogenic model of cerebral injury in pretreatment paradigm. Fifteen rabbits received a unilateral cryogenic brain lesion. Six animals received U83836E, and nine animals received vehicle only. Arterial blood gases, hematocrit, temperature, and mean arterial pressure were controlled in physiologic range throughout the experiment. Intracranial pressure was continuously monitored. Lesion size was determined by triphenyltertrazolium chloride staining and planimetric image analysis. Neutrophil aggregation and oxygen-free radical generation was assessed in whole blood by impedance aggregometry and simultaneous luminol-mediated chemiluminescence, respectively. Myeloperoxidase assay was used to determine neutrophil accumulation within the brain. Elevations in intracranial pressure from baseline values were significantly greater in the untreated (control) animals at all time points (p < 0.0001). Mean lesion size (% hemisphere +/- SEM) was significantly reduced in animals receiving U83836E (4.41 +/- 0.64, n = 6) when compared with controls (15.09 +/- 2.28, n = 9, p < 0.001). A reduction in both the release of oxygen free radicals and in neutrophil aggregation following the cryogenic injury in the U83836E animals correlated with a smaller lesion volume.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7473912 TI - Extraluminal, transluminal, and observational treatment for vertebral artery injuries. AB - Injury to the vertebral artery following penetrating trauma is rare and treatment is usually surgical ligation. Recent liberal use of angiography in the evaluation of penetrating neck trauma has identified increasing numbers of patients with this challenging injury. This report describes our recent experience in treating patients with vertebral artery injuries. The purposes of this study were (1) to review the outcome of our patients with vertebral artery injuries, and (2) to develop an approach for managing these patients. Sixteen patients were treated over a 9-year period. Three patients underwent emergent operative exploration for bleeding, three underwent transcatheter embolization alone, and ten were managed conservatively by close clinical observation. No deaths occurred. Ligation was performed for injuries discovered during neck exploration, however, bleeding was sometimes persistent despite proximal control. In our center, where radiological support is readily available, temporary control of bleeding by packing with hemostatic agents allowed subsequent transcatheter embolization of the injured artery. Pseudoaneurysms, arteriovenous fistulae, and extravasations discovered angiographically were usually managed by transcatheter embolization. Patients with vertebral artery narrowings or occlusions were managed by close clinical observation. PMID- 7473913 TI - The new epidemic in children: penetrating injuries. AB - OBJECTIVE: It is our hypothesis that there has been a dramatic increase in penetrating injuries in children. The purpose of this study is to verify this "new" epidemic in children and to note some of its characteristics. METHODS: We performed a 7-year retrospective review of the trauma registry at our urban pediatric (< 16 years of age) trauma center (UPTC). RESULTS: The percentage of admissions due to penetrating injuries at our UPTC has gradually risen over the past 7 years primarily due to gunshot wounds. In 1992 and 1993, compared to 1987 and 1988, the incidence of penetrating injuries has increased from 20 to 36% in all children and from 45 to 66% in the 12-to-15-year-old age group. CONCLUSION: Injuries due to penetrating trauma have, indeed, increased to epidemic proportions. PMID- 7473914 TI - A prospective study of surgeon-performed ultrasound as the primary adjuvant modality for injured patient assessment. AB - Ultrasound diagnostic imaging, having been used in Germany in the trauma setting for more than 15 years, has unique qualities that give it distinct advantages over other tests (DPL, CT), and is gradually gaining acceptance by surgeons in the United States. In this prospective study, experienced surgeon sonographers successfully used ultrasound as the primary adjuvant modality to detect hemoperitoneum and pericardial effusion in injured patients. The ultrasound evaluations of 371 patients demonstrated that in 65 patients with significant injuries, ultrasound detected 53, that is, had an 81.5% sensitivity and 99.7% specificity. They conclude that ultrasound should be the primary adjuvant instrument for the evaluation of injured patients because it is rapid, accurate, and is potentially cost-effective. PMID- 7473915 TI - Laparoscopy in 121 consecutive patients with abdominal gunshot wounds. AB - OBJECTIVE: The purpose of this study was to evaluate the sensitivity, specificity, and predictive value of diagnostic laparoscopy (DL) in a large group of stable patients with abdominal gunshot wounds (ABGSWs). DESIGN: This study was a prospective case series developed by management protocol. MATERIALS AND METHODS: In a 2 1/2-year period, DL was performed in 121 consecutive patients who were hemodynamically stable with ABGSWs and met protocol criteria. This represented 18% of all patients with ABGSWs seen in this period at the Ryder Trauma Center. The evaluation was conducted to determine peritoneal violation, the presence of intra-abdominal blood, and the need for exploration. MEASUREMENTS AND MAIN RESULTS: There were 42 (35%) positive and 79 (65%) negative DLs. In patients with positive DL, 39 (92.8%) had exploratory laparotomy. In this group, 32 (82%) had therapeutic laparotomy, 6 (15.4%) had nontherapeutic laparotomy, and 1 (2.5%) had a negative laparotomy. In this patient, DL was felt to be inadequate by the attending surgeon, although no penetration or intraperitoneal blood were present, and a negative laparotomy was done. This represents a failure rate of 0.8%. There were 3 (7.2%) positive DLs, in whom laparotomy was not performed. These patients had isolated nonbleeding liver injuries, and nontherapeutic laparotomy was successfully avoided. The negative DL group was divided into 47 patients (60%) with isolated ABGSWs, and 32 patients (40%) with associated injuries, mostly orthopedic and thoracic. There were no false-negative DLs and no delayed laparotomies in these 121 patients. There was no mortality in this study group. The sensitivity for peritoneal penetration was 100%, and the specificity was 98.7%. The positive predictive value was 97.6%, and the negative predictive value was 100%. In deciding on need for laparotomy (i.e., injury requiring repair), DL had a positive predictive value of 82%; more importantly, the negative predictive value was 100%. CONCLUSIONS: In stable patients with ABGSWs and questionable intra-abdominal injury, DL can be safely used. It is highly sensitive and specific. It can effectively reduce the incidence of negative and nontherapeutic laparotomies, and the overall morbidity and hospital stay in this group of patients. PMID- 7473917 TI - Computed tomography is inaccurate in estimating the severity of adult splenic injury. AB - Computed tomography (CT) is increasingly utilized in evaluation of adult splenic injury (SI). CT correlation with operative findings, CT relationship to successful nonoperative (NO) management, and CT reading reproducibility were examined. Records of patients > or = 15 years old admitted over a 3-year period were reviewed. Computed tomography scans were graded by two radiologists blinded to clinical results. Computed tomography scans were performed on 49 of 77 patients with SI. Eighteen underwent initial operation (OR) and 31 initial NO. Operative patients had higher Injury Severity Scores and Abdominal Abbreviated Injury Scale scores (p < 0.0001). Grade II readings predominated in the NO group (55%). Nonoperative management was successful for 9 grade III and 3 grade IV readings. Computed tomography matched OR grade in 10 readings, underestimated it in 18, and overestimated it in 6. Computed tomography missed SI in five patients. Radiologists disagreed on 9 of 45 (20%) scans. Computed tomography poorly predicted operative findings. Interobserver variability was common. SI management should not be based solely on CT severity. PMID- 7473916 TI - Management of blunt splenic trauma: computed tomographic contrast blush predicts failure of nonoperative management. AB - Nonoperative management of blunt splenic trauma is widely accepted; however, reported failure rates have ranged as high as 40%. There are few factors available to identify failures reliably. To characterize failures of nonoperative management better, we retrospectively reviewed 309 blunt splenic injuries treated at our level I trauma center over a 5-year period. Eighty-nine patients were initially managed nonoperatively (29%), and 12 patients failed this approach (13%). Upon review of the initial computed tomography scans, a hyperdense collection of contrast media in the splenic parenchyma, or "contrast blush," was noted in 8 of 12 (67%) patients who failed and in 5 of 77 (6%) of those who were successfully managed nonoperatively (p < 0.0001). These data suggest that the presence of a contrast blush is an important consideration when deciding the method for management of the splenic injury. If these results are confirmed in a prospective fashion, the failure rate of nonoperative management of blunt splenic trauma could be reduced by identification of the contrast blush. PMID- 7473918 TI - Adverse consequences of increased intra-abdominal pressure on bowel tissue oxygen. AB - OBJECTIVE: Demonstrate the effect that increased intra-abdominal pressure (IAP) has on visceral oxygen delivery and bowel tissue oxygenation (TPO2). METHODS: Six Duroch swine underwent abdominal insufflation with helium to pressures of 15 and 25 mm Hg for 1 hour. Animals were instrumented with a pulmonary artery flotation catheter to measure cardiac output and calculate systemic oxygen delivery. Fluorescence quenching optodes were implanted in the terminal ileum and the subcutaneous tissue of an axillary fold to measure bowel and systemic (control) tissue oxygen levels, respectively. RESULTS: Bowel tissue oxygen fell from 43 +/- 12 mm Hg at baseline to 31 +/- 12 mm Hg, with 15 mm Hg of abdominal pressure at 60 minutes. With 25 mm Hg IAP, bowel TPO2 fell from 24 +/- 12 to 12 +/- 8 mm Hg (p < 0.02). No change in axillary TPO2 was observed during either period of increased IAP. Cardiac output (CO), systemic oxygen delivery, and mixed-venous oxygen saturation (Svo2) also declined, although blood pressure and oxygen consumption remained constant. CONCLUSIONS: Increased IAP produces significant decreases in bowel submucosal TPO2 without similar changes in extra-abdominal (subcutaneous) TPO2. This decline is dependent on the extent and duration of the elevation in IAP. Readily accessible parameters, such as CO and Svo2, also decline with increased IAP and may be useful variables to monitor as relative indicators of bowel hypoperfusion and TPO2. PMID- 7473919 TI - Increased survival after major thermal injury: the effect of growth hormone therapy in adults. AB - BACKGROUND: Advances in the management of patients with major thermal injury have resulted in a progressive increase in survival rates. We report preliminary data evaluating the safety and potential efficacy of human growth hormone (HGH) administration in a high-risk population of burned patients. METHODS: From 1989 to 1993, 69 patients sustaining major burns (defined as patient age plus percentage of body surface area with deep second- and third-degree burns > or = 90) were evaluated. Patients routinely received anti-inflammatory pharmacotherapy including antioxidants, an endotoxin binder, and cyclooxygenase blockade. Half of the 54 patients who survived more than 7 days received HGH to enhance wound healing. Injury severity, morbidity, and mortality for patients receiving HGH was compared to the 27 patients not receiving HGH. RESULTS: For the entire population (n = 69), average age was 56 +/- 23 years, body surface area burned was 58% +/- 24%, and 30% sustained smoke inhalation. Actual mortality was 41%, significantly less than the more than 70% mortality rate predicted from reported outcome data. Patients receiving HGH were well matched with the group not receiving HGH with respect to extent of injury, burn management, pharmacotherapy, and in-hospital morbidity. Mortality of the patients receiving HGH was 11%, significantly less than the 37% mortality rate of the patients without HGH (p = 0.027). CONCLUSION: Compared to standard predictors of burn mortality our small patient group appears to have an improved survival rate, suggesting that the use of anti-inflammatory agents appears safe and potentially beneficial. Patients receiving HGH exhibited minimal drug-related complications and mortality rates were improved when this population was compared with both predicted mortality rates and a well-matched control population of concurrently treated patients. Prospective blinded trials are now necessary to confirm these findings in a larger patient group. PMID- 7473920 TI - Seroprevalence of human immunodeficiency virus, hepatitis B virus, hepatitis C virus, and rapid plasma reagin in a trauma population. AB - We evaluated the presence of human immunodeficiency virus (HIV), hepatitis B virus (HBV), hepatitis C virus (HCV), and rapid plasma reagin (RPR) among patients admitted to our trauma unit from April 15 to June 30, 1993. Of 984 patients tested, we found 255 (26%) had evidence of exposure to one or more of these agents: HIV, 4%; HBV, 20%; HCV, 14%; and RPR, 1%. Thirty-eight percent of patients had more than one positive serology, 75% of the HIV patients, 49% of the HBV patients, and 66% of the HCV patients. There was no difference between penetrating and nonpenetrating trauma with respect to any of the viruses. The risk factors for HIV-positive patients were non-White race, positive drug screen, positive alcohol screen, and city resident. Risk factors for HBV patients were non-White race, positive drug screen, and city resident. Risk factors for HBC patients were male sex, non-White race, positive alcohol screen, positive drug screen, and city resident. The risk of blood-borne infections in this group of patients is substantial. PMID- 7473921 TI - Surgical management of abdominal wall disruption after blunt trauma. AB - Abdominal wall disruption following blunt trauma is a rare but challenging injury, both in the acute and convalescent phases. The present report describes the recent experience with this injury at a single adult trauma center. In a 22 month period, nine patients with traumatic abdominal wall disruption were managed. Flank and anteroinferior abdominal wall defects were most common. Associated injuries included 6 patients with a pelvic fracture and 4 patients with rectosigmoid injuries. Immediate primary repair of the defect was attempted in seven cases at the time of trauma laparotomy, but was difficult and often unsuccessful because of the related tissue destruction. Delayed abdominal wall repair was performed in patients with symptomatic disability (n = 5) and, if required, restoration of intestinal continuity was performed at a separate operation before abdominal wall repair. Delayed repair with autogenous tissue included the use of tensor fascia lata, rectus femoris muscle, rectus abdominis fascia, and latissimus dorsi muscle. Reconstruction with prosthetic mesh was required in two patients. One early and one late recurrence occurred, resulting in reoperation. In conclusion, traumatic abdominal wall disruption represents a complex challenge for both general and plastic surgeons. The key to successful surgical management seems to be a delayed staged repair with autogenous tissue when feasible. PMID- 7473922 TI - Major trauma in Australia: a regional analysis. AB - This study was undertaken to evaluate the frequency, distribution, cause, pattern, and outcome of patients suffering from major trauma in the State of Victoria over a 1-year period. No previous study in Australia has attempted a comprehensive regional analysis of major trauma. All major trauma admissions resulting from blunt, penetrating, and burns injury were identified, and data collected from emergency departments and intensive care log books at 25 major metropolitan and rural hospitals from the January 3, 1992 to February 28, 1993 by onsite data collectors. The total number of patients admitted into the study was 2,944. There were 1,076 major trauma cases with an Injury Severity Score greater than 15 in a population of 4.2 million people. The type of injury was predominantly blunt (87.5%), with only a small percentage of penetrating injuries (6.4%) and burns (6%). Major trauma in pediatric cases is less common (132 cases). The most common causes of injury were road transport (56%) and falls (22%). The overall outcome of the group was favorable when compared with the Major Trauma Outcome Study group (Z = 1.4, M = 0.93, W = 0.52). There was an unexpectedly low number of patients suffering from major trauma. Outcome using Trauma and Injury Severity Score methodology was favorable when compared with North America. PMID- 7473924 TI - Tangential excision and skin grafting for ash burns of the foot in children: a preliminary report. AB - In India, isolated burns of the feet in children are common, but people seldom seek specialized treatment. These burns result from careless disposal of hot ash or embers after cooking or religious functions. Five children were treated for acute burns of the feet, with tangential excision and skin grafting. Early surgical treatment of these seemingly innocuous burns offers distinct advantages, i.e., avoidance of painful dressings, the need for reconstructive procedures, and reduced hospital stay. It also helps in early return to childhood activities and normal growth of the foot. A new concept of excising deep dermal burns of the "sole" is emphasized to preserve sensate unburned dermis. PMID- 7473925 TI - The role of toxic oxygen metabolites in a young model of thermal injury. AB - Production of oxygen-free radicals has been proposed as one pathophysiologic mechanism for postburn cardiac contractile dysfunction in adults. To examine this hypothesis in young subjects, we studied the cardiac effects of polyethylene glycol-superoxide dismutase (PEG-SOD) and PEG-catalase (PEG-CAT), each given as 20 U/g of body weight with fluid resuscitation (Parkland formula), after a third degree burn constituting 33% of the total body surface area in young (6- to 7-day old) guinea pigs (group 3, n = 12). Fluid-treated burns without scavenger therapy (group 2, n = 15) and sham burn controls (group 1, n = 15) were included. Animals were killed 24 hours postburn, and hearts were studied in vitro (Langendorff). Compared with sham burn controls, fluid-treated burns (group 2) had significant cardiac dysfunction as indicated by a lower peak systolic left ventricular (LV) pressure (LVP: 67 +/- 2 vs. 57 +/- 4 mm Hg, p = 0.01, mean +/- SEM), maximal rate of LV pressure development (+dP/dt max: 1169 +/- 45 vs. 988 +/- 45 mm Hg/second, p = 0.01), and fall (-dP/dt max: 1109 +/- 45 vs. 919 +/- 49 mm Hg/second, p = 0.01). In addition, LV function curves calculated for group 2 were shifted downward and to the right of those calculated for sham burn controls in the direction of contractile depression, p = 0.01. PEG-SOD/PEG-CAT treatment in burns did not significantly improve LVP (60 +/- 5 mm Hg), but scavenger therapy improved +/-dP/dt max values (1112 +/- 74 and 988 +/- 98 mm Hg/second, respectively).(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7473923 TI - Health care professional attitudes toward donation: effect on practice and procurement. AB - Failure to procure organs, tissues, and corneas for transplantation can be attributed to a number of factors. The existing literature is largely speculative concerning why health care professionals (HCPs) fail to approach the families of medically suitable donors and why requests for donation are not successful. This study is based on the direct examination of 1,797 HCP attitudes and knowledge about donation in conjunction with how HCPs performed when faced with a donor eligible patient. HCP attitudes, rather than knowledge, are more important to the successful procurement of organs. The HCPs with more positive attitudes about donation and their role in the procurement process were more likely to request donation. HCPs were more successful in obtaining consent to donation when they believed that the donation process would benefit the donor family and that their efforts to procure organs would be successful. These results indicate that educating HCPs about the donation process to make them more comfortable with it is crucial. PMID- 7473926 TI - Arterial gunshot injuries of the extremities: a South African experience. AB - This is a retrospective study of 173 patients with gunshot injuries of the major arteries of the extremities. A selective policy for the use of angiography was followed. The arterial repair was achieved by various means: primary end-to-end anastomosis, vein interposition graft, and polytetrafluoroethylene (Teflon) interposition grafts. Overall, there were nine amputations in the lower limb. Ninety-eight percent of the patients had a palpable pulse on discharge. There were 3 preoperative deaths, 1 intraoperative death, and 5 postoperative deaths (overall perioperative mortality, 3.5%). We conclude that the results of vascular extremity gunshot injuries are satisfactory when standard methods of management are used. Morbidity and mortality can be further reduced by prompt admission to appropriate centers. PMID- 7473928 TI - Axillary artery injuries secondary to anterior dislocation of the shoulder. AB - We describe a case report of an axillary artery injury after recurrent anterior dislocation of the shoulder. Review of 22 reported cases reveals that 27% are recurrent dislocations, 86% occurred in patients older than 50, 86% of the injuries are in the third part of the axillary artery, and 68% presented with an axillary mass. PMID- 7473927 TI - Blunt gallbladder injuries: presentation of twenty-two cases with review of the literature. AB - Blunt injuries to the gallbladder are very uncommon, whereas penetrating gallbladder trauma occurs more frequently. Over the last 19-year period through 1994, 22 cases with blunt gallbladder trauma were treated. Avulsion (partial or complete) and contusion were observed in 10 cases each (45%). Perforation of the gallbladder and traumatic cholecystitis with necrosis were seen in one case each. Associated intra-abdominal trauma was present in 100% of the cases and accounted for both mortalities. Gallbladder injury was diagnosed after celiotomy, except for one case that was diagnosed and treated nonoperatively. Based on the experiences gained herein and an extensive review of the literature, a classification scheme for gallbladder trauma and its treatment is presented. PMID- 7473929 TI - Venovenous bypass as an adjunct for the management of a retrohepatic venous injury in a child. AB - We report the successful utilization of venovenous bypass in the case of an 8 year-old-boy who suffered a major hepatic injury with a retrohepatic venous injury after a kick by a horse. Initial nonoperative management was successful, but a delayed hemorrhage required operation. Vascular access was obtained via the peripheral circulation, and venovenous bypass was performed without systemic heparinization. Venovenous bypass improved vascular control and allowed the repairs to be performed in a relatively bloodless field, while not compromising venous return. PMID- 7473930 TI - Delayed hemothorax resulting from stab wounds to the internal mammary artery. AB - Massive delayed hemothorax as a result of stab wounds to the internal mammary artery have not recently been reported. A patient who died after such an injury prompted a 10-year retrospective review. During that time period, 88 patients with stab wounds to the chest were identified. Thirty-five of these involved injuries to the parasternal region. This region is described as being located below the clavicles, between the midclavicular lines, and above the costal margins. We identified 5 patients with internal mammary artery injuries: 4 had massive delayed bleeding and 2 died. Our current recommendations for managing stable patients with parasternal stab wounds include maintaining a high index of suspicion, intensive care monitoring, and placement of chest tubes for monitoring. Should delayed bleeding occur, a prompt thoracotomy is mandatory. PMID- 7473931 TI - Distal innominate artery transection and cervical spine injury. AB - Distal innominate artery injury secondary to blunt trauma had not previously been reported. We present a case of distal innominate artery transection associated with a cervical spine dislocation. A mechanism for this unusual vascular injury is proposed. An organized diagnostic and treatment plan is emphasized. PMID- 7473932 TI - Pseudoaneurysms of the intraparenchymal splenic artery after blunt abdominal trauma: a complication of nonoperative therapy and its management. AB - The presentation, diagnosis, and successful management of posttraumatic pseudoaneurysms of the intraparenchymal splenic artery after nonoperative therapy in an adult patient is described. Pseudoaneurysm formation of the intraparenchymal splenic artery is a rare complication of traumatic splenic injury, which is a potential mechanism of delayed splenic rupture and demonstrates the importance of follow-up in the nonoperative therapy of blunt injury to the spleen. PMID- 7473933 TI - Shotgun wounds and pellet emboli: case reports and review of the literature. AB - Shotguns produce extensive tissue damage when victims are injured at close range. Arterio-arterial and veno-venous pellet emboli may result when major vessels or their tributaries are injured. Management of the pellet embolus is dictated by whether it is symptomatic or asymptomatic. Regardless of symptoms, the embolus is a sign of possible major vascular or venous tributary injury. We recently managed two cases of pellet emboli produced by close range shotgun wounds. This report describes our management of these injuries and collectively reviews the past 30 years of literature on shotgun wounds with arterio-arterial and veno-venous pellet emboli. Recommendations for management are suggested from collating the cases reported in the literature. PMID- 7473934 TI - Small bowel laceration from a penetrating extraperitoneal gunshot wound: a case report. AB - Direct kinetic energy transfer from the path of a bullet may cause intraperitoneal injury even though the path of the bullet remains extraperitoneal. A 45-year-old man sustained a single through-and-through gunshot wound to the abdomen. The path of the bullet appeared superficial but clinically the patient had an acute abdomen. An exploratory laparotomy revealed no penetration of the peritoneum, but did reveal a perforation and contusion to the jejunum. To our knowledge this is the first reported case of a penetrating extraperitoneal gunshot wound with a blast injury to the small bowel. PMID- 7473935 TI - Posttraumatic inferior vena caval thrombosis: case report and review of the literature. AB - This report describes an autopsied case of inferior vena caval thrombosis that occurred secondary to blunt trauma in a 21-year-old man. Fourteen days after the injury, he died of massive thromboembolism of the main pulmonary artery. Implications of this case and a review of the literature are presented. PMID- 7473936 TI - Nosocomial meningitis caused by multiresistant enterococcus as a life-threatening complication of pelvic injury: case report. AB - Bacterial meningitis after pelvic trauma has never been described. We recently treated a patient who developed, during the course of his hospitalization, multiresistant enterococcal meningitis after severe pelvic injury, including a comminutive sacral fracture. Dural tear may have been the main factor leading to secondary infection of the cerebrospinal fluid. Treatment with intravenous continuous infusion of vancomycin plus rifampin, associated with closed subarachnoid drainage, resulted in a complete cure. Therapeutic cerebrospinal fluid levels of vancomycin were obtained only during the first 8 days of treatment. Use of glycopeptides in meningitis and the role of cerebrospinal fluid drainage are discussed. Physicians should be aware of the diagnosis and therapeutic features of this life-threatening complication. PMID- 7473937 TI - Biliary tract injury following blunt abdominal trauma: case reports. AB - This report reviews three cases of biliary tract injury following blunt abdominal trauma. During diagnostic evaluation, computerized tomography failed to delineate the injuries, but endoscopic retrograde cholangiopancreatography (ERCP) aided in the diagnosis in two patients and pre-operative biliary stent placement facilitated operative intervention. Surgical exploration was required to fully characterize the injury in all three patients. PMID- 7473938 TI - Candiduria as an early marker of disseminated infection in critically ill surgical patients. PMID- 7473939 TI - Electrical burns associated with head injuries. PMID- 7473940 TI - Morbidity and mortality of rib fracture. PMID- 7473941 TI - The comparison of injury severity instrument performance using likelihood ration and ROC curve analyses. PMID- 7473942 TI - Thermodilution evaluation of right ventricular performance. PMID- 7473943 TI - Delayed presentation of splenic artery pseudoaneurysms following blunt abdominal trauma. PMID- 7473944 TI - The Sickness Impact Profile as a tool to evaluate functional outcome in trauma patients. AB - Because the ultimate goal of trauma care is to restore injured patients to their former functional status, reliable evaluation of functional status is needed to assess fully the effectiveness of trauma care. We hypothesized that the Sickness Impact Profile (SIP), a widely used measure of general health status, would be a useful tool to evaluate the long-term functional outcome of trauma patients and that the SIP would identify unexpected problems in the recovery process and groups of patients at high risk for long-term disability. A prospective cohort of 329 patients with lower extremity fractures admitted to three level I trauma centers were interviewed using SIP at 6 and 12 months postinjury. Patients with major neurologic injuries were excluded. Overall SIP scores and each of the component subscores may range from 0 (no disability) to 100. In this series, the mean overall SIP was 9.5 at 6 months and 6.8 at 12 months, compared with a preinjury baseline of 2.5. At 12 months, 52% of patients had no disability (SIP 0 to 3), 23% mild disability (4 to 9), 16% moderate disability (10 to 19), and 9% severe disability ( > or = 20). Disability was widely distributed across the spectrum of activities of daily living, including physical functioning (mean score of 5.5), psychosocial health (mean score of 5.5), sleeping (mean score of 10.0), and work (mean score of 21.0). The SIP scores did not correlate with Injury Severity Score.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7473945 TI - Emergency department care and hospitalization as predictors of disability. AB - Duration of complete and partial disability for work, school, and home activities and activities of daily living during the first 18 months after injury were compared for 2,043 emergency department (ED) patients and 151 hospitalized patients from 22 northwestern Vermont communities who received their initial medical care for injury at the Medical Center Hospital of Vermont and were subsequently interviewed. Larger proportions of hospitalized patients than ED patients had any disability or prolonged disability. During almost all time frames, and even among patients who still had ongoing disability at 18 months, the majority of persons with disability had required ED treatment only. These data suggest that, based on disability, ED patients should not necessarily be considered to have merely minor injuries. PMID- 7473946 TI - Increased levels of circulating interleukin-8 in patients with large burns: relation to burn size and sepsis. AB - Large burns are followed by significant trauma-induced immunomodulation, and activated neutrophils can be demonstrated in the circulation of burn patients shortly after injury. Interleukin-8 (IL-8) is a recently described molecule with neutrophil activating properties and in the present study we have measured the concentration of this cytokine in plasma from 27 patients with large burns during hospitalization using an enzyme linked immunosorbent assay (ELISA). The mean patient plasma concentration of IL-8 at admission was about 60 times higher than that of healthy controls. Furthermore, patients with total body surface area burn of more than 40% had significantly higher IL-8 concentrations in plasma than patients with smaller burns. For patients without serious infectious complications, the IL-8 concentration fell gradually after injury, whereas in patients with complicating sepsis a second peak of IL-8 was demonstrated. Thus, the increased IL-8 concentrations seem to be related to burn size and to have a role in the pathophysiology of sepsis in patients with large burns. The large amounts of circulating IL-8 following thermal injury may contribute to the strong and sustained activation of neutrophils reported earlier in patients with large burns. PMID- 7473947 TI - Effects of trauma on leukocyte intercellular adhesion molecule-1, CD11b, and CD18 expressions. AB - BACKGROUND: During traumatic injury, a multitude of events, including ischemia, may cause leukocyte adhesion and margination. In this study, alterations of surface receptors involved in leukocyte adhesion were studied in traumatized patients. In an attempt to discern the role of hypoxia, additional experiments were conducted in which normal human leukocytes were subjected to hypoxic stress in vitro. METHODS: Venous blood was obtained from 10 trauma patients within 2 hours of blunt injury (mean Injury Severity Score of 17 +/- 8) and from 8 normal volunteers (controls). Leukocytes were isolated from patients and controls. To assess the effect of hypoxia, normal leukocytes were placed in hermetically sealed environments containing 100% nitrogen. All leukocytes were labeled with phycoerythrin- or fluorescein-bound monoclonal antibodies to intercellular adhesion molecule-1 (ICAM-1), or to integrins CD18 and CD11b. Receptor concentration was measured by flow cytometry. Results were expressed as percentage of receptor-positive cells (%) and mean fluorescence channel units, which directly correlate with monoclonal antibody cell surface density. Significance of differences was tested by analysis of variance/Kruskal-Wallis test. RESULTS: Compared with the normal controls, circulating leukocytes obtained from traumatized patients showed decreased expression of ICAM-1, CD11b, and CD18 2 hours after injury. In contrast, normal leukocytes exposed to hypoxic stress in vitro exhibited a marked increase in CD11b and CD18 expression and no change in ICAM-1 expression. CONCLUSIONS: Leukocytes obtained from traumatized patients showed a significant decrease in cell surface expression of adhesion receptors. This phenomenon is unlikely to be a direct consequence of hypoxia alone, because exposure to isolated hypoxia in vitro actually increased expression of CD11b and CD18. PMID- 7473948 TI - Effect of surgical trauma on splenocyte and peritoneal macrophage immune function. AB - Although previous studies have shown that simple laparotomy produces a depression in peritoneal macrophage (Mphi) antigen presentation capacity, it remains unknown whether the adverse effects of laparotomy are limited to peritoneal Mphi or whether such an insult also affects splenocyte immune function. To study this, mice were anesthetized and a 1-inch midline abdominal incision was made, followed by abdominal closure. At 2 and 24 hours after the surgical procedure, the animals were killed, splenocyte cultures established and stimulated for 48 hours with concanavalin A (2.5 micrograms/mL), while peritoneal macrophage cultures were stimulated with LPS (10 micrograms/mL). The proliferative capacity of the splenocytes, as well as their ability to release interleukin-2 and interleukin-3, was markedly decreased at 2 as well as 24 hours after laparotomy. Furthermore, the release of interleukin-6 by splenic and peritoneal macrophages from animals that underwent laparotomy were also significantly depressed at both 2 and 24 hours. These results support the concept that surgical stress in the form of midline laparotomy per se is sufficient to produce a significant impairment in cell-mediated immunity, thus setting the stage for increased incidence of postoperative complications. PMID- 7473949 TI - Asymptomatic deep venous thrombosis in the trauma patient: is an aggressive screening protocol justified? AB - The incidence and sequelae of deep venous thrombosis (DVT) in trauma patients are unclear because the majority of patients who develop DVT are asymptomatic. This study evaluated the incidence, risk factors, and efficacy of prophylaxis for DVT in trauma patients over a 5-year period. Trauma patients at high risk for DVT were evaluated biweekly with lower extremity venous duplex scans. The DVT prophylaxis was instituted on admission with low-dose heparin therapy and pneumatic compression. The incidence of asymptomatic DVT identified by duplex screening was 10% (45 of 458); one pulmonary embolus occurred. Five variables were significant from bivariate and multiple logistic regression analysis: age (p = 0.005), Injury Severity Score (p = 0.005), length of stay (p = 0.004), Trauma and Injury Severity Score (p = 0.01), and spinal injury (p = 0.014). This analysis documents that trauma patients with these risk factors are at increased risk for the development of asymptomatic DVT, despite prophylaxis, and warrant surveillance with venous duplex sonography. PMID- 7473950 TI - Soluble thrombomodulin increases in patients with disseminated intravascular coagulation and in those with multiple organ dysfunction syndrome after trauma: role of neutrophil elastase. AB - OBJECTIVE: Our goal was to investigate the role of soluble thrombomodulin (TM) and neutrophil elastase in patients with trauma. DESIGN: This study is a prospective case-control study. MATERIALS AND METHODS: Forty-seven trauma victims, 14 with disseminated intravascular coagulation (DIC), 5 with multiple organ dysfunction syndrome (MODS), and 28 control patients without DIC or MODS were the participants. Soluble TM and neutrophil elastase (elastase-alpha1 proteinase inhibitor complex) were measured on the day of the injury, and on the first, third, and fifth days after admission. The results of these measurements and demographic data were compared among the groups, and correlations between the soluble TM and the neutrophil elastase were examined. The DIC patients were classified into subgroups of survivors (n = 5) and nonsurvivors (n = 9), and the changes of the soluble TM between the subgroups were then studied. MEASUREMENTS AND MAIN RESULTS: A high incidence of DIC patients encountered MODS complications (12 of 14, 86%). The DIC patients had higher Injury Severity Scores (ISSs) than the other patients. The levels of soluble TM and neutrophil elastase significantly increased on the day of admission in the patients with DIC and also in those with MODS (p < 0.05 vs. control patients) and continued to show markedly high values until the fifth day of admission in the patients with DIC. In the DIC patients, the levels of soluble TM were higher in the nonsurvivors than in the survivors (p < 0.05 on the third and the fifth days of admission). In all patients, there was weak but statistically significant correlation between peak levels of soluble TM and ISS (r2 = 0.125, p < 0.025). Comparison of the levels of soluble TM and neutrophil elastase in the patients with DIC or MODS demonstrated an excellent correlation (r2 = 0.718 and r2 = 0.714, respectively). CONCLUSIONS: Soluble TM as a novel endothelial cell injury marker increases in patients with DIC and also in those with MODS after trauma. Neutrophil elastase may be involved in the pathogenesis of the injury. Soluble TM is a marker of the severity of injury and is a good predictor of MODS. PMID- 7473951 TI - Bile and bilirubin excretion in relation to hepatic energy status during hemorrhagic shock and hypoxemia in rabbits. AB - OBJECTIVE: We investigated the relation between in vivo hepatocyte excretion of bile and bilirubin and hepatic energy status in rabbit models of hemorrhagic shock and hypoxemia. DESIGN: Randomized animal study. MATERIALS AND METHODS: After creation of a total biliary fistula, hemorrhagic shock with mean pressure of 50 mm Hg (10 rabbits) or hypoxemia with Pao2 at 35 mm Hg (8 rabbits) was induced for 60 minutes. We determined bile flow, excretion of bilirubin and total bile acids, the plasma level of bilirubin, and arterial ketone body ratio, which reflects hepatic mitochondrial function. MEASUREMENTS AND MAIN RESULTS: Both the hemorrhagic shock and the hypoxemic models showed decreases in bile flow and excretion of bilirubin and total bile acids as well as increase in the plasma level of bilirubin in association with decreases in the hepatic energy charge and the arterial ketone body ratio. CONCLUSIONS: Bile flow and the excretion of bilirubin were correlated with the hepatic energy status. PMID- 7473952 TI - Venovenous bypass for major hepatic and caval trauma. AB - Severe trauma to the liver and juxtahepatic cava require vascular isolation of the lesion. In addition to the techniques of vascular clamping and atriocaval shunting, venovenous bypass is a viable alternative to repair such injuries. We herein report its use in five consecutive patients. The technique obviates the hemodynamic sequelae of uncompensated caval occlusion and the technical dangers of atriocaval shunting. PMID- 7473953 TI - Liver injury as a model of uncontrolled hemorrhagic shock: resuscitation with different hypertonic regimens. AB - Using a standardized liver injury model of uncontrolled hemorrhage, we tested the effect of different hypertonic solutions on mortality, blood pressure, intra abdominal bleeding, and circulating blood volume. After liver injury, rats were randomized to 4 groups: lactated Ringer's (LR, n = 10), Isosal (ISO, n = 10), hypertonic saline (HS, n = 10), and hypertonic sodium acetate (HA, n = 10). In all resuscitation groups, 4 mL/kg was infused at a rate of 0.4 mL/min. Blood volume was evaluated both directly and by estimation. Mortality was highest after HA resuscitation (40%) and lowest after HS resuscitation (0%), but this difference was not significant. Blood pressure was significantly higher after HS resuscitation, and this difference was sustained for 4 hours. The HA resuscitation did not increase blood pressure compared with LR resuscitation. Intraperitoneal blood volume was significantly higher with HS (25.5 +/- 0.7 mL/kg) and HA (26.8 +/- 1.2 mL/kg) than with LR (22.5 +/- 0.4 mL/kg). The HA resuscitation led to a significantly larger drop from baseline values of estimated terminal circulating blood volume than LR resuscitation. Nonparametric analysis combining survival time and directly measured change in blood volume demonstrated a significant advantage to HS, compared with LR. HA and HS resuscitations increased bleeding from uncontrolled solid viscus injury. The HS resuscitation restored blood pressure better than the other hypertonic solutions and maintained circulating blood volume in spite of increased bleeding. The HA and ISO resuscitations did not exhibit any advantage over LR in resuscitation of solid viscus injury. PMID- 7473954 TI - Traumatically injured Jehovah's Witnesses: a sixteen-year experience of treatment and transfusion dilemmas at a level I trauma center. AB - Traumatically injured Jehovah's Witnesses pose difficult management problems because of their refusal to accept blood transfusions. This retrospective review of all inpatient traumatically injured Jehovah's Witnesses at a level I trauma center over the past 16 years revealed 77 patients with 92% blunt and 8% penetrating injuries. The primary physician was aware of their unique religious status in only 32% of cases. Transfusion was performed in only 4 (5.2%) cases even though it was desired by the physician in 11 (14%) cases. One transfusion was performed against the patient's will. One minor was transfused using a court order. Two transfusions were performed in the trauma room before the patients' religious status was known. Major changes in therapeutic plans were made as a result of the patients' Jehovah's Witness status in 10 cases (13%). Early knowledge of the patient's religious status is essential to optimize patient care. PMID- 7473955 TI - Safe and normothermic massive transfusions by modification of an infusion warming and pressure device. AB - A fluid-warming pressure infusion device (H-500/H25i, Level 1) was modified to meet the demands for safe, normothermic, and effective massive transfusions. By incorporating an autoventing 40 microns filter (Pall AV-SP), which was originally manufactured for use in an extracorporal circulation, the risk of accidental air embolism can be eliminated. Feasibility and efficiency of this model were tested in ten patients. The mean volume transfused and infused (packed red cells and colloids) was 6750 mL (SD +/- 2519) during a mean period of acute volume resuscitation of 55 minutes (SD +/- 30). Calculated flow rates averaged 140 mL/min, which were sufficient to stabilize all but one patient. This patient subsequently died because of uncontrollable surgical bleeding. Body temperature remained stable with a minimal mean drop of -0.3 degrees C. Supplying the Level 1 warming and pressure device with a Pall AV-SP filter allows for safe, effective, and demand-adapted massive transfusions in a large number of trauma patients at a reasonable cost. PMID- 7473956 TI - Large-volume intraoperative peritoneal lavage with an assistant device for treatment of peritonitis caused by blunt traumatic rupture of the small bowel. AB - The benefits of large-volume intraoperative peritoneal lavage (IOPL), with an assistant lavage device, were evaluated retrospectively in 114 patients with peritonitis caused by blunt traumatic rupture of the small bowel. Postoperative complications caused by infection were a major problem after rupture of the small bowel (46 of 114, 39.4%). Both prolongation of the interval between injury and laparotomy and rupture of the lower part of the small bowel were risk factors for postoperative complications caused by infection. Large-volume IOPL (25.2 +/- 2.1 L) with an assistant lavage device reduced the rate of complications caused by infection from 30 of 58 (51.8%) to 15 of 56 (26.8%). The volume used for IOPL was closely related to the occurrence of postoperative complications resulting from infection. No complications from infection occurred in patients who received lavage with of 28.3 +/- 2.7 L of saline, whereas complications occurred in those patients treated with a smaller volume of lavage fluid (18.0 +/- 2.5 L). Large volume IOPL should be considered in patients with blunt rupture of the small bowel who are at risk for infection, and the assistant device for IOPL may be useful for such treatment. PMID- 7473957 TI - Iliofemoral venous injuries: an indication for prophylactic caval filter placement. AB - Prophylactic placement of vena caval filters is recommended in trauma patients at high risk for pulmonary embolism (PE). We present a group of patients with iliofemoral venous trauma, and subsequent complications of deep venous thrombosis (DVT) and PE. Of twelve patients with iliac or common femoral venous injuries, seven underwent primary repair. All received DVT/ PE prophylaxis with mini-dose heparin and/or sequential compression hose. In spite of this, two patients suffered DVT, one patient had DVT and PE and one patient had clinical evidence of PE but did not undergo confirmatory testing. Three patients underwent prophylactic caval filter placement without complication. The DVT/PE complication rate in this small group was at least 43% (3 of 7). Patients with repaired iliofemoral venous injuries represent a high risk subset for DVT/PE and prophylactic caval filter placement is recommended. PMID- 7473958 TI - Diagnosis and management of minor head injury: a regional multicenter approach in Italy. AB - Two series of patients admitted to the hospital after a minor head injury were collected in two different periods (1985 and 1989) in a regional hospital with a 24-hour computed tomography (CT) service, but without a neurosurgical unit. In 1988, a regional protocol on the management of patients with minor head injury (based on the presence of skull fractures in adults and on clinical parameters in children) was adopted. There was a 21% reduction in hospital admission in adults, and the number of skull x-ray films performed in children decreased significantly (p < 0.01). A more liberal use of CT examinations in asymptomatic patients with skull fractures produced an earlier identification of patients with extradural hematomas who were sent to neurosurgery before clinical deterioration with good results. Detection of cerebral contusions was clinically less important. Based on the availability of CT scanners in our area and on the results of our study, we have proposed new guidelines in management of minor head injury. The CT scans are obtained in patients with a Glasgow Coma Scale score of 13 or less. Skull x-ray films are obtained in patients older than 10 years with a Glasgow Coma Scale score of 14/15. If a fracture is found, the patient is sent to the nearest regional center for CT examinations. Children younger than 10 years are sent to a regional hospital with 24-hour CT availability for clinical observation or other indicated studies. PMID- 7473959 TI - Fractures of the floor of the anterior cranial fossa. AB - We treated 28 patients with anterior cranial fossa floor fractures. Computed tomography (CT) scans adjusted to bone density disclosed three fracture types: (1) penetrating fractures through the orbita or ethmoid sinus; (2) simple or multiple linear fractures; and (3) extensive comminuted anterior cranial fossa floor fractures. Thirteen patients underwent emergent surgery for treatment of open depressed fractures (most common in type 3 fractures), for foreign bodies (in type 1 fractures), and for optic canal decompression. Large dural lacerations were always present in patients with type 3 fractures, and repairs were made with dural substitutes. Only one patient developed postoperative cerebrospinal fluid leakage. Nine (32%) of the patients in our series had visual involvement, but visual acuity recovered or improved in six patients. Our study shows that initial neuroradiologic evaluation with CT scans is important in patients with frontobasal fractures, and that secure dural repair during primary operation helps prevent cerebrospinal fluid leakage. PMID- 7473960 TI - Outcome of tibial shaft fractures with severe soft tissue injury treated by unreamed nailing versus external fixation. AB - MATERIALS: Between 1987 and 1991, 114 fresh tibial shaft fractures with severe soft tissue injury, which met the inclusion criteria, were treated and retrospectively reviewed. There were 48 cases in the unreamed nail (unreamed tibial nail (UTN)) group and 66 cases in the external fixation (EF) group. METHODS: Soft tissue injury was classified by the Gustilo and Tscherne methods. There were 18 AO type A, 56 type B, and 40 type C fractures. There were no significant difference in fracture type, soft tissue injury, age, and additional injury between the groups. Outcome was assessed using the Karlstrom score after 28 months. RESULTS: There were a significantly higher number of reoperations in the EF group. Mean time to bony healing was 25.8 weeks. In the EF group, there was a 26% rate of pin tract infection, whereas 19% of cases had bolt breakages in the UTN group. In the UTN group, 40% had good results, compared with 27% in the EF group. In the UTN group, there were significantly fewer ante- and recurvatum deformities of more than five degrees. CONCLUSIONS: Treatment of tibial fractures by UTN, compared with EF, gave a lower reoperation rate and better functional outcome. PMID- 7473961 TI - Does lung contusion affect both the traumatized and the noninjured lung parenchyma? A morphological and morphometric study in the pig. AB - Isolated unilateral lung contusion (LC) was induced in 12 pigs to determine the pathophysiological role of LC in the high mortality after multiple injury. The Horovitz quotient, pulmonary vascular resistance, mean pulmonary artery pressure, mixed venous oxygen consumption, inspiratory pressure and compliance were significantly decreased in the LC group as compared to controls. The number of polymorphonuclear granulocytes, the microvascular permeability of albumine and the Wilhelmy balance as determined by bronchoalveolar lavage were significantly increased after lung contusion. As indicators of a systemic reaction we found elevated plasma levels of the terminal complement complex and decreased levels of the complement factor 3a after LC. The morphological assessment revealed changes such as those encountered during the early phase of adult respiratory distress syndrome, with granulocyte sticking, endothelial cell adhesion and transendothelial diapedesis. Morphometric analysis demonstrated a significant decrease in alveolar diameter in both the injured and the contralateral lung due to impaired surfactant surface activity. A distinct increase in septal diameter, related to edema and caused by increased microvascular permeability, was found in the injured lungs. These findings emphasize that LC leads to a generalized impairment of the entire lung, which may lead to progressive lung failure. PMID- 7473962 TI - Comparative analysis of radiographic interpretation of orthopedic films: is there redundancy? AB - Cost containment is becoming the watchword in today's medical care environment. In an effort to determine possible areas of unnecessary patient cost secondary to redundant services, we decided to compare prospectively interpretations of plain orthopedic films by radiologists and orthopedists. Without performing a physical examination of the patient, orthopedic surgical attendings and radiology attendings independently read 507 consecutive radiographic studies of acute orthopedic injuries sustained by 438 patients. All readings were dictated, and the reports were reviewed by the senior author and statistically analyzed. The cost of the radiologists' readings was computed. Analysis of the two types of readings showed that both were highly sensitive and very specific, and that there was no statistically significant difference (p = 1.0) between them. The average cost of the radiologists' readings in the local area was approximately $16,100. There was no fee for orthopedic interpretations in this study. The authors conclude that because the two interpretations were accurate and not statistically different, interpretation of orthopedic films by a radiologist seems to be an unnecessary expense. PMID- 7473963 TI - Unnecessary use of pelvic x-ray in blunt trauma. AB - Pelvic radiography in blunt trauma patients is routinely used in most trauma centers. The purpose of this review was to evaluate the ability of physical examination alone to detect pelvic fractures. Among patients with blunt trauma admitted to the University Hospital del Valle in Cali, Colombia, over a 3-month period, 608 adult patients, with hemodynamic stability, without spinal involvement, and with a Glasgow Coma Scale score greater than 10 were evaluated. All patients had physical examination, pelvic x-ray and adequate follow-up. Fifty nine (9.7%) patients had pelvic fractures, with 57 of these fractures detected by physical findings. The remaining two patients had stable fractures that required no treatment. After careful analysis, we conclude that a negative physical examination following blunt trauma has a negative predictive value of 99% probability in excluding pelvic fracture, provided that the patient is not a child, is not in coma, is hemodynamically stable without evidence of blood loss, and has no spinal cord injury. A selective use of pelvic x-ray in patients with blunt trauma is a cost-effective policy. PMID- 7473964 TI - Bier's block: a change of injection site. AB - Bier's block or intravenous regional anesthesia is a well-proven technique that is useful in the traumatic setting to provide upper limb anesthesia in the manipulation of distal radial fractures. In this situation, the traditional injection site in the dorsum of the hand can present difficulties with subsequent plaster application and with venous access caused by swelling or pain. One hundred patients were randomly allocated into two groups of 50. The first group was injected into the dorsum of the hand and the second into the antecubital fossa, in each case on the injured side. Effectiveness of anesthesia at both the fractures site and the cuff was assessed using a visual analog scale. Results analyzed using Student's t test, showed no difference in anesthesia between the two groups. In addition, there were fewer technical problems associated with venous access and subsequent plaster application in the antecubital fossa group. PMID- 7473965 TI - Pediatric cervical-spine immobilization: achieving neutral position? AB - This study was designed to evaluate prospectively the ability of current spine immobilization devices to achieve radiographic-neutral positioning of the cervical spine in pediatric trauma patients. All trauma patients who required spinal immobilization and a lateral cervical spine radiograph were included in the study. A lateral cervical spine radiograph was obtained while the child was immobilized. The Cobb angle (C2-C6) was measured using a handheld goniometer. The method of immobilization, age at injury, and Cobb angle were compared. One hundred and eighteen patients with an average age of 7.9 years were enrolled. The majority were males (71%). The most frequent mechanisms of injury included motor vehicle accidents (35%) and falls (32%). The average Glascow Coma Scale score was 14. Although 31% of the children complained of neck pain, 92% were without neurologic deficits. The Cobb angles ranged from 27 degree kyphosis to 27 degree lordosis, and only 12 of the patients presented in a neutral position (0 degrees). Greater than 5 degrees of kyphosis or lordosis was observed in 60% of the children. Thirty-seven percent of the patients had 10 degrees or greater angulation. The most frequent methods of immobilization included a collar, backboard, and towels (40%), and a collar, backboard, and blocks (20%), but these techniques provided < 5 degrees kyphosis or lordosis in only 45% and 26% of the children respectively. No single method or combination of methods of immobilization consistently placed the children in the neutral position.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7473966 TI - Croatian experience in the treatment of 629 urogenital war injuries. AB - During the war in Croatia, from April 1991 until November 1993, 33,468 wounded persons were reported, and 24,865 of them were hospitalized. Out of the total number of the hospitalized persons 588 wounded suffered 629 injuries of the urogenital system. This study reports on the experience in the treatment of such injuries at 17 urological and surgical institutions. PMID- 7473967 TI - Severe skiing injuries: a retrospective analysis of 361 patients including mechanism of trauma, severity of injury, and mortality. AB - All ski accident patients requiring an inhospital treatment at our institute from 1984 to 1992 (n = 2,053) were analyzed retrospectively. The incidence, pattern, and severity of the injuries, as well as the 30-day mortality comparing two time slots (1984 to 1988 and 1989 to 1992) were analyzed for the 361 cases classified as "serious" injuries according to the following definitions and groupings: group 1, multiple trauma (Injury Severity Score > or = 18) and patients with multiple fractures (n = 179; group 2, abdominal or thoracic single trauma (Abbreviated Injury Scale (AIS) score > or = 2, n = 58); and group 3, isolated head injuries (AIS score > or = 2, n = 124). Serious injuries were observed in 19% in the first period compared to 16% in the second period (not significant). Two hundred thirty eight of 361 patients injured themselves by just falling, while 117 collided with some sort of obstacle like other skiers (45), trees or rocks (27), posts, pylons, barriers (20), and moving objects (25) such as piste machines, ski lifts, automobiles, and in one case a helicopter. Six skiers were caught on the ski run by an avalanche. Comparing the two time periods, trauma circumstances did not change significantly. The number of most severe head injuries (AIS score > or = 4) increased from 11.6 to 19.3% (p < 0.05). The overall mortality increased from 2 to 7% (p < 0.05). Group 2 had the best prognosis with no mortality, while group 3 was rated worst with 8%. Skiing remains a major source of serious trauma in winter resort areas.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7473968 TI - Relevance of carbohydrate-deficient transferrin as a predictor of alcoholism in intensive care patients following trauma. AB - Every second traumatized patient is a chronic alcoholic. Chronic alcoholics are at risk due to an increased morbidity and mortality. Reliable and precise diagnostic methods for detecting alcoholism are mandatory to prevent posttraumatic complications by adequate prophylaxis. The patient's history, however, is often not reliable, and conventional laboratory markers are not sensitive or specific enough. The aim of this study was to investigate whether carbohydrate-deficient transferrin (CDT) is a sensitive and specific marker to detect alcoholism in traumatized patients. One hundred and five male traumatized patients or their relatives gave their written informed consent to participate in this institutionally approved study. All patients were transferred to the intensive care unit after admission to the emergency room, followed by surgical treatment. Diagnostics included an alcoholism-related questionnaire, conventional laboratory markers (mean corpuscular volume, gamma-glutamyltransferase, aspartate aminotransferase, and alanine aminotransferase), and CDT sampling (microanion exchange chromatography, turbidimetry, and radioimmunoassay, respectively). Only patients in whom a reliable history could be obtained were included. Alcoholism was diagnosed if the patients met the Diagnostic and Statistical Manual of Mental Disorders criteria for chronic alcohol abuse or dependence. The administration of fluids before CDT sampling was carefully documented. Patients did not differ significantly regarding age, Trauma and Injury Severity Score, and Acute Physiology and Chronic Health Evaluation score. The sensitivity of the CDT research kit was 70% and of the commercially available kit CDTect was 65%. Early sampling in the emergency room and before administration of large volumes of fluid increased the sensitivity to 83% for the CDT research kit and 74% for CDTect, respectively.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7473969 TI - Acute trauma alters morphine clearance. AB - Trauma is accompanied by changes in liver perfusion and acute phase proteins. Such changes have the potential to alter drug metabolism. There are few studies describing drug disposition in acute trauma. We determined the pharmacokinetics of an intermediate extraction drug, morphine, in trauma patients. Nine patients with an Injury Severity Score (ISS) > or = 16 were studied within 48 hours of trauma. Morphine 5 mg was given intravenously and serial blood and urine samples were drawn to derive pharmacokinetic parameters. Alpha acid glycoprotein (AAG) levels were determined. Total morphine clearance (CL) and volume of distribution (Vss) were decreased compared to established literature values. Area under the curve (AUC) and terminal half-life (T 1/2 alpha) were increased. AAG levels were higher than reference range. Elimination half-life was increased. The decrease we observed in Vss may be attributed to increased binding of morphine by AAG, which is increased after trauma as in our patients. Decreased clearance and increased half-life of an intermediate extraction drug may be explained by increased protein binding, decreased liver blood flow, and reduced hepatocellular function. Decreased clearance of the magnitude observed in these patients could result in drug accumulation. Better understanding of the effects of trauma on the pharmacokinetics of low, high, and intermediate extraction drugs will prevent excessive or suboptimal drug dosing. PMID- 7473970 TI - Acute repair of aorta-innominate vein fistulas. AB - Penetrating wounds of the central vasculature are highly lethal. Ten cases of aorta-left innominate vein fistulas have been reported in the past, but most have presented late after injury. We report three successfully managed cases that had a short interval between injury and repair, and where we used cardiopulmonary bypass with total circulatory arrest. Preoperative arteriography facilitated planning the operative approach. The ready availability of complete radiologic and surgical resources at a trauma center were responsible for the successful outcome of these highly lethal central vascular injuries. PMID- 7473971 TI - Temporary vascular continuity during damage control: intraluminal shunting for proximal superior mesenteric artery injury. AB - "Damage control" is an accepted technique for the treatment of the patient with exsanguinating injuries to the abdomen and intraoperative coagulopathy. We describe the use of an intraluminal shunt to maintain temporary vascular continuity of the superior mesenteric artery during rewarming and correction of coagulopathy in the intensive care unit following a gun shot wound to the abdomen. Successful complex reconstruction was achieved at definitive laparotomy using an autogenous vein interposition graft. PMID- 7473972 TI - Traumatic injury to the azygous vein: case report. AB - Injury to the azygous vein is rare in blunt chest trauma. A review of the world's literature revealed only 10 cases of trauma-related azygous vein injury. We report another patient who survived azygous vein rupture secondary to thoracic rib injury. With prompt recognition, emergency thoracotomy and proper therapy, patients with azygous vein injury have a high survival rate. PMID- 7473973 TI - Carotid artery pseudoaneurysm and pellet embolism to the middle cerebral artery following a shotgun wound of the neck. AB - Arterial missile embolism is a rare complication of penetrating vascular trauma. We report a case of middle cerebral artery pellet embolism and delayed appearance of a carotid artery pseudoaneurysm following a shotgun wound of the neck. The pseudoaneurysm was repaired. Because the patient had no associated neurologic deficits, the pellet embolus was left within the patient middle cerebral artery. He remains well 4 years after injury. A selective approach to the management of a pellet embolus to the middle cerebral artery based on clinical signs or symptoms and status of arterial patency is recommended. In addition, several principles are suggested to improve the reliability of arteriography for shotgun wounds of the neck. PMID- 7473974 TI - Hypopituitarism secondary to transfacial gunshot wound. AB - Hypopituitarism secondary to penetrating head trauma is extremely rare, and its diagnosis may be delayed for several years. We present a patient who developed hypopituitarism secondary to a transfacial gunshot wound and who experienced damage to the hypophysis secondary to transmission of energy and bullet fragments. The importance of a computerized tomographic scan of the head in facial gunshot wound is discussed. PMID- 7473975 TI - Diagnosis of coronary artery dissection following blunt chest trauma by transesophageal echocardiography. AB - How to differentiate relevant from trivial cardiac injury in blunt chest trauma has been an ongoing debate. In a 32-year-old victim of a motorcycle crash, the electrocardiographic pattern of an acute anterior wall myocardial infarction was identified as being due to a dissection, after an intimal flap in the proximal left anterior descending artery was noted on transesophageal echocardiography. PMID- 7473976 TI - Tension pneumopericardium following blunt injury. AB - Pneumopericardium caused cardiac tamponade in a patient who was struck in the chest by a motor vehicle. Subxiphoid pericardial window and pericardial drainage successfully treated this condition. Diagnosis of this rare form of tamponade depends on clinical examination supported by chest radiographic findings. PMID- 7473977 TI - Late repair of chest deformity secondary to traumatic manubriosternal disruption: case report. PMID- 7473978 TI - Lethal Waterhouse-Friderichsen syndrome in posttraumatic asplenia. AB - We herein report a case of fulminant lethal Waterhouse-Friderichsen syndrome in an elderly female patient seven years after posttraumatic splenectomy. In contrast to various reports, this patient had not been vaccinated against Streptococcus pneumoniae, Neisseria meningitidis, or Haemophilus influenzae, respectively, although infections with these microorganisms are known to cause the main lethal diseases in asplenic patients. Again, we recommend obligatory vaccinations against the mentioned bacteria for it is known that this decreases the risk of fatal septic events in these patients. To optimize prevention, it is imperative to vaccinate patients undergoing splenectomy before discharge from hospital. PMID- 7473979 TI - Compartment syndrome in the critically injured following massive resuscitation: case reports. AB - Compartment syndrome of the extremities is usually associated with locally mediated events such as ischemia or injury. Three cases are presented of patients who did not sustain direct extremity trauma, but who developed compartment syndrome following massive fluid resuscitation. Global ischemia/reperfusion may be responsible for the observed morbidity. Patients undergoing large volume resuscitation should be closely monitored for this entity. PMID- 7473980 TI - Calcific myonecrosis following compartment syndrome: report of two cases, review of the literature, and recommendations for treatment. AB - Calcific myonecrosis of lower extremity muscles has been identified as an uncommon late sequela of posttraumatic compartment syndromes or ischemic myonecrosis. Previous reports have detailed a small number of cases with similar clinical and radiographic features. Two cases of calcific myonecrosis diagnosed more than 30 years following trauma to an extremity are presented. Treatment consisted of serial debridement of all infected and necrotic tissue followed by wound closure using a rectus abdominis muscle microvascular transplant in one patient and delayed primary closure in the other. Both patients have resumed active lifestyles with no recurrence of drainage or infection. Aggressive debridement and definitive soft tissue coverage are the mainstay of effective treatment for this entity. PMID- 7473981 TI - Neuroleptic malignant syndrome in a trauma patient. AB - Neuroleptic malignant syndrome is a rare complication of antipsychotic medications. This paper describes the case of a trauma patient who received haloperidol and droperidol for severe agitation, developed neuroleptic malignant syndrome, and was treated successfully with dantrolene sodium, and also provides a review of the literature. PMID- 7473982 TI - Intracranial hypertension and adult respiratory distress syndrome: usefulness of tracheal gas insufflation. AB - The management of increased intracranial pressure (ICP) in patients with an associated acute lung injury is difficult. High levels of PaCO2 as tolerated for permissive hypercapnia are deleterious for cerebral circulation. In such circumstances, tracheal gas insufflation (TGI), which was recently proposed to reduce PaCO2, may be of benefit. We report the cases of two patients with severe adult respiratory distress syndrome and head trauma complicated with elevated ICP. The introduction of TGI decreased PaCO2 by 17 and 26%, decreased ICP, and increased calculated cerebral perfusion pressure. We conclude that TGI could be added to a pressure-targeted strategy of ventilatory management when severe adult respiratory distress syndrome was associated to an intracranial hypertension. PMID- 7473983 TI - Rhabdomyolysis following spinal cord injury: case report. AB - Rhabdomyolysis is commonly seen in association with multisystem trauma. We report an unexpected case of rhabdomyolysis and subsequent renal insufficiency after spinal cord injury unassociated with acute muscular injury. Immobilization, in conjunction with mild systemic hypoperfusion, was the suspected cause. Due to the likelihood of muscle catabolism, the difficulties of diagnosing soft tissue injury, and the impact of any decrease in renal reserve in patients with spinal cord injuries, surveillance for laboratory evidence of rhabdomyolysis in patients with neurologically complete spinal cord injuries appears warranted. PMID- 7473984 TI - Candida mediastinitis and septic shock following occult esophageal perforation in a patient with posttraumatic quadriplegia. AB - Unexplained septic shock was ultimately shown to be caused by Candida mediastinitis after perforation of the cervical esophagus by a dislodged cervical methylmethacrylate construct in a 25-year-old patient with traumatic quadriplegia. Communication between the prevertebral abscess and pleural space further led to the formation of a esophageal-pleural-cervical fistula. Despite antibiotics, surgical removal of the construct, and drainage of the esophagus and mediastinum, the patient died from refractory shock and respiratory failure. PMID- 7473985 TI - Femoral head necrosis after intertrochanteric fracture: a case report after synthesis with Ender nails. AB - A case of femoral head necrosis in a patient who suffered an intertrochanteric fracture, treated with Ender nails 1 year before, is presented. The rarity, and possible mechanisms (reduction and type of osteosinthesis) of necrosis are discussed. PMID- 7473986 TI - Low-velocity gunshot fractures of the radius and ulna: case report and review of the literature. AB - This report describes a case of right radial-ulnar fracture with bone loss, and left olecranon fracture with radial nerve palsy resulting from a small caliber, low-velocity gunshot. Initial wound debridement and extraskeletal fixation were done to fix the radial-ulnar fracture. Wound debridement, bullet removal, and tension band wiring were performed on the left forearm. Definitive operations with rigid plate fixation and full segmental iliac bone grafting of the right radial-ulnar fracture were done 2 months later. Left radial nerve palsy resolved 9 months after the index injury. The fractures were fully united and both forearm functions were in normal range 3 years postoperatively. Implications of this case and a review of the literature are presented. PMID- 7473987 TI - Delayed embolization of a shotgun pellet from the chest to the middle cerebral artery. AB - Embolization of metallic missiles into the cerebral circulation is a rare occurrence. Most of the cases reported were due to gunshot wounds and shotgun wounds to the neck and face. Embolization from injuries sustained to the chest are extremely rare. We report a case of delayed pellet embolization to the middle cerebral artery resulting from a shotgun injury to the left chest. PMID- 7473988 TI - Rupture of the deep femoral artery from blunt trauma. AB - Massive hemorrhage from the deep femoral artery is an uncommon entity in the setting of blunt extremity trauma without femur fracture. A case of deep femoral artery injury causing massive hemorrhage treated by angiographic embolization is reported. In this patient, persistent unexplained hypotension warranted angiographic analysis of a pelvic fracture. Because of a swollen right thigh, the negative pelvic angiogram was extended to include the lower extremity, confirming the diagnosis of a ruptured branch of the deep femoral artery. Bleeding was controlled with embolization that promptly resolved the patient's hemodynamic instability. The salient feature in common with previous reported cases of deep femoral artery injury was hemodynamic instability beyond accountable blood loss. We recommend angiographic analysis with radiological and surgical intervention in the setting of thigh swelling without femur fracture and unexplained hypotension. This management strategy was well tolerated, and the patient received minimal transfusions. PMID- 7473989 TI - Blunt carotid artery injury: devastating consequences of undetected pseudoaneurysm. AB - A case of delayed embolization of a traumatic pseudoaneurysm of the right common carotid artery, resulting in fatal cerebral infarction, is reported. This case emphasizes the importance to detect occult lesions to the carotid arteries after blunt neck injuries and to treat aggressively pseudoaneurysms located upstream to a cerebral vessel to prevent embolic complications. PMID- 7473990 TI - Steal phenomenon in a traumatic carotid-cavernous fistula. AB - A patient with head injury presented with computed tomography findings of a diffuse severe infarction of the left cerebral hemisphere in which the cerebral hemodynamics can be evaluated by transcranial Doppler sonography. Serial angiograms revealed a carotid-cavernous fistula, with a complete steal phenomenon. The unusual complication of a traumatic carotidcavernous fistula is discussed. PMID- 7473992 TI - Diverse burdens to health care causes by weapon violence. PMID- 7473991 TI - Internal mammary artery injury, anterior mediastinal hematoma, and cardiac compromise after blunt chest trauma. AB - A rare case of blunt chest trauma resulting in internal mammary artery hemorrhage and cardiac tamponade is presented. Thoracotomy revealed anterior mediastinal hemorrhage but no pericardial hematoma. The significance of chest wall vessel hemorrhage as a cause of widened mediastinum is reiterated. The importance of accurate angiographic assessment and vigilant care of victims of blunt chest trauma who present with a widened mediastinum is emphasized. PMID- 7473993 TI - A stake through the heart of colostomy. PMID- 7473994 TI - Trauma data bases. PMID- 7473995 TI - On timeless principles in changing times. PMID- 7473996 TI - Nonoperative salvage of computed tomography-diagnosed splenic injuries: utilization of angiography for triage and embolization for hemostasis. AB - OBJECTIVES: The aims of this study were to determine if angiographic findings can be used to predict successful nonoperative therapy of splenic injury and to determine if coil embolization of the proximal splenic artery provides effective hemostasis. METHODS: Splenic injuries detected by diagnostic imaging between 1981 and 1993 at a level I trauma center were prospectively collected and retrospectively reviewed after management by protocol that used diagnostic peritoneal lavage, computed tomography (CT), angiography, transcatheter embolization, and laparotomy. Computed tomography was performed initially or after positive diagnostic peritoneal lavage. Angiography was performed urgently in stabilized patients with CT-diagnosed splenic injuries. Patients without angiographic extravasation were treated by bed rest alone; those with angiographic extravasation underwent coil embolization of the proximal splenic artery followed by bed rest. RESULTS: Patients (172) with blunt splenic injury are the subject of this study. Twenty-two patients were initially managed operatively because of associated injuries or disease (11 patients) or because the surgeon was unwilling to attempt nonoperative therapy (11 patients) and underwent splenectomy (17 patients) or splenorrhaphy (5 patients). One hundred fifty of 172 consecutive patients (87%) with CT-diagnosed splenic injury were stable enough to be considered for nonoperative management. Eighty-seven of the 90 patients managed by bed rest alone, and 56 of 60 patients treated by splenic artery occlusion and bed rest had a successful outcome. Overall splenic salvage was 88%. It was 97% among those managed nonoperatively, including 61 grade III and grade IV splenic injuries. Sixty percent of patients received no blood transfusions. Three of 150 patients treated nonoperatively underwent delayed splenectomy for infarction (one patient) or splenic infection (two patients). CONCLUSIONS: (1) Hemodynamically stable patients with splenic injuries of all grades and no other indications for laparotomy can often be managed nonoperatively, especially when the injury is further characterized by arteriography. (2) The absence of contrast extravasation on splenic arteriography seems to be a reliable predictor of successful nonoperative management. We suggest its use to triage CT-diagnosed splenic injuries to bed rest or intervention. (3) Coil embolization of the proximal splenic artery is an effective method of hemostasis in stabilized patients with splenic injury. It expands the number of patients who can be managed nonoperatively. PMID- 7473997 TI - Functional status following orthopedic trauma in young women. AB - OBJECTIVE: To evaluate the general health status and sexual function of women following serious orthopedic injury. METHODS: Women aged 16-44 who were treated at a level I trauma center between 1986 and 1992 for a fracture to the pelvis or lower extremity were interviewed by telephone. The interview included the Medical Outcomes Study 36-Item Short Form Health Survey (SF-36) supplemented by questions about sexual function. The SF-36 is a measure of outcome from the respondent's point of view and consists of 36 items representing eight health concepts. RESULTS: Of 289 eligible women, 233 (81%) were interviewed (123 pelvic fracture; 110 lower extremity fracture). Their mean Injury Severity Score was 17.9. Compared to age- and gender-standardized norms, study patients as a group scored significantly worse (lower scores) on all dimensions of the SF-36 except mental health (p < 0.05). Of the women interviewed, 45% reported feeling less sexually attractive due to their injury, and 39% reported a decrease in sexual pleasure. Women who reported arthritis that was attributed to their fracture had significantly poorer health outcomes than study subjects who did not. The most significant predictor of deviations from SF-36 norms was the presence of one or more comorbid chronic conditions. CONCLUSION: The results underscore the importance of considering comorbidities when evaluating health outcomes following major trauma. In addition, the relatively high rates of reported change in sexual function after injury argue for more attention to these issues in both clinical practice and outcomes research. PMID- 7473998 TI - Functional outcome of patients with unstable pelvic ring fractures stabilized with open reduction and internal fixation. AB - An unstable pelvic ring fracture represents a severe injury and is associated with high morbidity and mortality. Little data are available assessing the long term functional limitations, including disability, in a patient with an unstable pelvic ring fracture. The purpose of this study was to describe the impairment and functional outcome (disability) for patients with unstable pelvic ring fractures managed with open reduction and internal fixation (ORIF). Disability was measured at a minimum of 1 year postinjury using the Sickness Impact Profile (SIP), a measure of the health-related quality of life as perceived by the patient. Of the 230 consecutive patients with a pelvic ring fracture, 54 had unstable fractures requiring ORIF; 48 patients were available at a 1 year follow up. The follow-up roentgenograms confirmed an osseous union and an anatomic alignment of the pelvis. Thirty-seven (77%) of the patients had mild disability (total SIP < 10); 11 (23%) of the patients had moderate disability (SIP > 10) at 1 year. Of the patients who were employed preinjury, 76% were employed 1 year postinjury; 62% had returned to full time work and 14% had returned with job modification. Of the 7 patients who had been in school, 6 had returned full time and 1 student returned part time. Mean SIP scores for subcategories were: physical health = 6.8 +/- 9.4, psychosocial health = 7.4 +/- 12.7, work = 17.6 +/ 25.5, home management = 8.3 +/- 13.0, ambulation = 10.7 +/- 13.7, and mobility = 5.3 +/- 13.0. Despite the magnitude of the bony injuries, the majority of patients with unstable pelvic ring fractures managed with ORIF had mild disability 1 year postinjury; the majority of the patients had returned to work. PMID- 7473999 TI - Permissive hypercapnia in trauma patients. AB - The use of a normal tidal volume in patients with progressive loss of alveolar airspace may increase inspiratory pressure and overdistend remaining functional alveoli. Permissive hypercapnia (PH) is a ventilator management technique that emphasizes control of alveolar pressure, rather than PCO2. The purpose of this study was to determine if the use of PH is associated with an improved outcome from adult respiratory distress syndrome (ARDS). Over a 2-year period, 39 trauma patients were treated for ARDS. Permissive hypercapnia was used in 11, and the remaining patients were treated conventionally. Demographics and risk factors were well matched in PH patients and controls. The duration of mechanical ventilation was greater in PH patients [49.2 +/- 15.2 vs. 20.8 +/- 10 days (p < 0.01)]. Survival was also greater in the PH group [91% vs. 48% (p < 0.01)]. A reduction in intensity of mechanical ventilation is associated with a prolongation of ventilatory support and an improved outcome from ARDS. PMID- 7474000 TI - Permissive hypercapnia as a ventilatory strategy in burned children: effect on barotrauma, pneumonia, and mortality. AB - OBJECTIVE: To document the incidence of barotrauma, pneumonia, and respiratory death associated with a mechanical ventilation protocol based on permissive hypercapnia in pediatric burn patients. DESIGN: Retrospective review. MATERIALS AND METHODS: Patients were managed using a mechanical ventilation protocol based on permissive hypercapnia, tolerating moderate (pH > 7.20) respiratory acidosis to keep inflating pressures below 40 cm H2O. MAIN RESULTS: Over a 2.5-year interval, 54 burned children (11% of 495 acute admissions) with an average age of 6.5 years (range 5 weeks to 17 years), average burn size of 44% (range 0 to 98%), and median burn size of 46% required mechanical ventilatory support for an average of 12.5 days (range 1 to 56 days). Inhalation injury was diagnosed in 34 (63%) of the children and 72% percent were admitted within 24 hours of injury. Overt barotrauma occurred in 5.6% of the patients, pneumonia in 32%, and respiratory death in 0%. CONCLUSIONS: A conventional ventilation protocol based on permissive hypercapnia is associated with acceptable rates of barotrauma and pneumonia. The low incidence of respiratory death associated with this strategy suggests that it also minimizes ventilator-induced lung injury. PMID- 7474001 TI - Neurogenic pulmonary edema in fatal and nonfatal head injuries. AB - Impaired pulmonary function is a frequent but poorly understood complication of acute head injury (HI). A potential early contributor to the pulmonary dysfunction seen in HI patients is neurogenic pulmonary edema (NPE). We hypothesized that NPE would occur early after HI and that it would have a continuum of clinical severity depending on the severity of the HI and associated intracranial hypertension. A large autopsy data base and inpatient HI data base were used to search for cases of NPE. Patients in the autopsy data base were stratified according to injury type and whether they died at the scene or within 96 hours of injury. There were significant (p < 0.0001, analysis of variance) elevations in lung weights in patients dying at the scene and within 96 hours from HI, compared with those dying from other noncentral nervous system injuries. No other organs studied showed significant weight increases. The incidence of NPE in isolated HI patients dying at the scene was 32%. In patients with isolated HI dying within 96 hours, the incidence of NPE was 50%. We found an inverse correlation (r = 0.62; p < 0.0014) between the initial cerebral perfusion pressure and the PaO2/FIO2 ratio despite a normal-appearing chest x-ray film. We conclude that NPE occurs frequently in HI patients. The process of edema formation begins early in the clinical course and is isolated to the lung.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7474002 TI - Burn-induced nitric oxide release in humans. AB - Nitric oxide (NO) generation in a series of 20 burn patients was studied with a novel anion exchange high-performance liquid chromatographic method for the simultaneous determination of nitrite (NO2-) and nitrate (NO3-), the stable metabolic endproducts of NO. The NO values within our survivor group (n = 17) were significantly altered at days 1, 6, and 12 postburn in contrast to controls (n = 23) (p < or = 0.03). NO2- values were significantly depressed in both plasma and urine, whereas NO3- values were significantly elevated in contrast to control values (p < or = 0.03). The ratio of NO2-:NO3- was significantly lower for burn patients versus controls in both plasma and urine (p < 0.01). The NO generation seemed in part to be dependent on the percentage of total body surface area burn, most dramatically elevated in patients with burns of 10 to 40% total body surface area. In subjects who did not survive beyond 36 hours postinjury because of irreversible shock (n = 3), the production of NO was significantly depressed in contrast to survivors and controls (p < 0.0001). However, the NO2-:NO3- ratio (0.001) was relatively unchanged, with reflection of a global depression in NO formation with no change in the individual component release. Burn injury resulted in an increased release/production of NO that in the first postburn week is maximally elevated immediately postinjury. NO release, although decreased at day 6 relative to the day 1 values, remained elevated into the second week postinjury when there was evidence for a further increase in NO production. The enhanced NO3- formation may well result from NO reacting with oxygen-free radicals counteracting superoxide anion-induced destruction of tissue, thereby potentially functioning as a protectant molecule. PMID- 7474003 TI - Release of anti-inflammatory mediators after mechanical trauma correlates with severity of injury and clinical outcome. AB - Excessive synthesis of proinflammatory cytokines [tumor necrosis factor (TNF) alpha and interleukin (IL)-1 beta] after trauma has been correlated with poor outcome. Recently, naturally occurring inhibitors of TNF-alpha and IL-1 beta have been characterized such as soluble TNF receptors (sTNFRs) and IL-1 receptor antagonist (IL-1ra). The present study was undertaken to determine whether injury results in a rise of circulating sTNFRs and IL-1ra. If so, whether plasma levels of these anti-inflammatory mediators correlate with severity of injury and clinical outcome of these patients. Injured patients (n = 213) showed significantly increased sTNFR and IL-1ra plasma levels throughout the observation period of 14 days, compared with healthy volunteers (n = 127). Patients with severe injury (Injury Severity Score > 16 points) revealed higher levels (p < 0.05) of sTNFRs and IL-1ra than patients with minor trauma (Injury Severity Score < or = 16 points). Patients who died from injury demonstrated increased (p < 0.05) sTNFR p55 and IL-1ra plasma levels, compared with survivors. Thus, anti inflammatory mechanisms are activated after trauma dependent on severity of injury. Because increased plasma levels of anti-inflammatory reacting proteins portended poorly for patient survival, these mediators may contribute to prediction of outcome after severe injury. PMID- 7474004 TI - Time course of the early pituitary-adrenal and metabolic responses to accidental injury. AB - We have made serial measurements of the plasma adrenocorticotrophic hormone (ACTH), cortisol, glucose, lactate, glycerol, nonesterified fatty acid (NEFA), and beta-hydroxybutyrate concentrations in 14 recently injured patients. We took 6 to 9 blood samples at about 20-minute intervals from each patient within the first 3 hours after accidental injury of moderate severity (Injury Severity Score of 9 to 13). Plasma ACTH varied greatly both with time and between patients; in the majority, it fell throughout the sampling period and in the others, the pattern varied. The changes in plasma cortisol were mostly appropriate to those in ACTH. Plasma glucose and glycerol did not change systematically with time. In contrast, plasma lactate fell, and plasma NEFA, beta-hydroxybutyrate, and the NEFA:glycerol ratio rose, throughout the sampling period. Analysis of variance for these data showed that time after injury made a contribution that was highly significant, although smaller than that arising from differences between patients. PMID- 7474005 TI - Primary repair of colon injuries: a prospective randomized study. AB - Due to the results of a 6-year experience with civilian penetrating colon injuries at Mount Carmel/Grace Hospital, in Detroit, Michigan, which had favored primary repair of colon injuries, a prospective randomized study was performed. Seventy-one patients with penetrating colon injuries were entered in a prospective randomized study. Forty-three patients were treated with primary repair or resection and anastomosis, and 28 patients were treated with diversion. The average Penetrating Abdominal Trauma Index score was 25.5 for the primary repair and 23.4 for the diversion groups. The majority of injuries as assessed by the Colon Injury Score (CIS) for the primary repair group were grades 2 (58%) and 3 (28%). The diversion group predominantly had grades 2 (64%) and 3 (25%). There was no significant difference between the two groups. There were 8 (19%) patients with colon and noncolon-related complications in the primary repair group, and 10 (36%) patients with colon, noncolon, and colostomy-related complications in the diversion group. In addition, there were 2 (7%) patients with complications following colostomy reversal. Independent risk factors for adverse outcomes were compared and used to calculate the probability for adverse outcomes with respect to the mode of treatment. The probability for adverse outcomes was statistically greater in the diversion group. An analysis was also made within the primary repair group comparing the subgroups of primary repair with, and without, resection of colon. It appears that the primary repair with resection of colon may have fewer complications; however, this conclusion is based on a statistically insufficient sample size. The authors contend that primary repair or resection with anastomosis is the method of choice for treatment of all penetrating colon injuries in the civilian population despite any associated risk factors for adverse outcomes. PMID- 7474006 TI - Use of echocardiography to detect occult cardiac injury after penetrating thoracic trauma: a prospective study. AB - One hundred five hemodynamically stable patients with penetrating thoracic trauma were prospectively evaluated for occult cardiac injury. All patients underwent transthoracic echocardiography (ECHO) and subxiphoid exploration (SXE). Those with positive SXE underwent exploration. Results are calculated based on operative findings of significant injuries. For the entire group, SXE was more sensitive than ECHO (sensitivity, 100% vs. 56%; specificity, 92% vs. 93%; and accuracy, 92% vs. 90%, respectively). The SXE missed no injuries in this group. Eight of 17 patients, however, with positive SXEs had insignificant injuries at sternotomy. Seven patients had positive ECHO, but no significant injury. The ECHO missed four significant injuries. When comparing SXE with ECHO in patients without hemothorax, however, sensitivity (100% vs. 100%), specificity (89% vs. 91%), and accuracy (90% vs. 91%) were comparable between SXE and ECHO. We conclude that ECHO has significant limitations in identifying serious cardiac injuries in patients with hemothorax. For hemodynamically stable patients without hemothorax, ECHO missed no significant injuries and may be an acceptable diagnostic option for detecting significant cardiac trauma in patients with injuries in proximity to the heart. PMID- 7474007 TI - Surgical indications in low lumbar burst fractures: experiences with Anterior Locking Plate System and the reduction-fixation system. AB - A study of 14 surgically treated patients with low lumbar burst fractures (L3 to L5) is reported. Initially, 13 patients presented with incomplete neurologic deficits below the level of injury. The follow-up period ranged from 24 to 40 months, with an average of 30 months. In group 1, seven patients underwent one stage anterior decompression, fusion, and Anterior Locking Plate System (ALPS) fixation. In Group 2, seven patients had posterior surgery with decompression, portero-lateral fusion, and a short segment fixation, one above and one below the injuried level, with the reduction-fixation transpediculate system. Indications for one-stage anterior surgery consisted of patients with incomplete neurologic deficits, segmental kyphotic deformity caused by loss of anterior vertebral height (< 50%), or canal encroachment < 50%. When there is significant radiologic evidence of lamina fractures, displaced or greenstick type, or injuries that occurred at the L5 level, the posterior approach was undertaken initially. This may be followed by anterior surgery if satisfactory vertebral height restoration or canal clearance was not achieved. Five dura tears (36%) were noted during surgery: two tears in group 1 and three tears in group 2. There were two patients in group 2 (29%) who had screws broken; however, none in group 1 demonstrated implant failure. In the low lumbar region, the step-off ALPS plate is preferable to the straight plate because of the increasing size of the vertebral body toward the caudal area. A low profile and easy application were its advantages. However, the ALPS could not be distracted or compressed anteriorly. PMID- 7474008 TI - Prediction of rib fracture injury outcome by an artificial neural network. AB - Outcome-based therapy is becoming the standard for assessing patient care efficacy. This study examines the ability of an artificial neural network to predict rib fracture injury outcome based on 20 intake variables determined within 1 hour of admission. The data base contained 580 patient records with four outcome variables: Length of hospital stay (LOS), ICU days, Lived, and Died. A 522-patient training set and a 58-patient test set were randomly selected. Nine networks were set up in a feed-forward, back-propagating design with each trained under different initial conditions. These networks predicted the test set outcome variables with an accuracy as high as 98% at the 80% testing level. Internal weight matrix examination indicated that age, ventilatory support, and high trauma scores were strongly associated with both ICU days and mortality. Being female, injury severity, and injury type were associated with increased LOS. Smoking and rib fracture number were low-level predictors of the four outcome variables. PMID- 7474009 TI - Triage in an established trauma system. AB - OBJECTIVE: The goal of this study was to determine patient and injury characteristics that predict undertriage and overtriage. DESIGN: This study was a retrospective analysis of admissions for acute injury. MATERIALS AND METHODS: All admissions for acute injuries in a 2 1/2-year period were included (N = 26,025). ICD-9 clinical modification codes were converted to Injury Severity Scores. MAIN RESULTS: Seventy-nine percent of severely injured patients were admitted to level I trauma centers. Severely injured patients admitted to other hospitals (undertriage) were more likely elderly (odds ratio = 5.44) and less likely had multisystem injuries (odds ratio = 0.55). One-fourth of patients with minor injuries were admitted to level I trauma centers (overtriage). Overtriaged patients were more likely intoxicated, obese, or had an injury to the head or face. CONCLUSIONS: In a developed trauma system, severely injured elderly trauma patients (especially females) are at risk for undertriage. The characteristics of patients at risk for overtriage reflect the difficulties of prospective out-of hospital triage. PMID- 7474010 TI - General surgeons and the Advanced Trauma Life Support course: is it time to refocus? AB - OBJECTIVE: The aim of this study was to assess Advanced Trauma Life Support (ATLS) training status of general surgeons, its perceived utility, and its relation to clinical trauma practice. METHODS: A national sample of 1300 general surgeons was surveyed by mail about trauma training, ATLS status, trauma call, and confidence in clinical trauma care abilities. RESULTS: Response rate was 61%. Respondents most commonly (67%) felt they learned a great deal about trauma care in residency training; 13% responded similarly regarding ATLS. Course participation within 4 years of the survey was reported by 33% of respondents. Nearly 75% of those not taking the course cited primary reasons related to relevance (30%), redundancy (29%), and credentialing (15%). Inaccessibility, inconvenience, and cost were lesser factors. Of those expressing extreme confidence with trauma resuscitation, 40% had taken ATLS; 15% of those expressing a lesser degree of confidence had taken ATLS. CONCLUSIONS: The ATLS course represents a standard of initial trauma care education in which only one-third of surgeons report current participation. Many view ATLS as not relevant or useful, yet take trauma call. To ensure standard education and patient care, an ATLS course curriculum specifically geared to the general surgeon should be developed and made a mandatory component of residency training or a requirement for board certification and trauma call credentialing. PMID- 7474011 TI - Adequacy of hospital discharge data for determining trauma morbidity patterns. AB - To determine the adequacy of hospital discharge data for describing the nature and severity of multiple trauma, injury data coded from full hospital records were compared with injury data coded from discharge summaries for 83 plane crash survivors admitted to 14 different hospitals. Full hospital records indicated that 33% sustained intracranial, 39% thoracic, and 27% abdominal/pelvic injuries. Thirty-five percent had spinal fractures, 40% upper limb fractures, and 70% lower limb fractures. Discharge summary data missed 52% of the intracranial injuries, 28% of the thoracic injuries, and 30% of the spinal fractures, yet only missed 5% of the lower limb and 15% of the skull, pelvic, and upper limb fractures. These differences point to concerns regarding the use of discharge data for determining the nature, severity, and sequelae of injuries. The observed underreporting of neurological injury suggests a need to re-evaluate discharge data systems for measuring brain and spinal cord injury patterns, particularly given the increasing reliance on these systems for health policy and programmatic decision making. PMID- 7474012 TI - An analysis of Hospital Discharge Index as a trauma data base. AB - STUDY OBJECTIVE: To document the validity of a Hospital Discharge Index (HDI) as a data base on injured patients. DESIGN: Patient information in trauma registries was compared with information in HDI. POPULATION: Injured patients admitted to trauma centers. METHODS: Patients in HDI were crossmatched with individuals in one or two trauma registries using deterministic matching techniques. Agreement regarding the presence and severity of injury was assessed. RESULTS: A comprehensive trauma registry from a level I trauma center and HDI agreed on the presence of an injury in each of 6 body regions over a range of kappa values from 0.17 to 0.71. The severity of injury score assigned by the two data bases demonstrated agreement over a range of intraclass correlation values from 0.12 to 0.82. CONCLUSION: HDI provides adequate information concerning injury for the majority of hospitalized patients, but was primarily limited by incomplete information. Efforts to improve HDI should focus on guidelines for data abstraction. PMID- 7474013 TI - The Injury Impairment Scale as a measure of disability. AB - Scores on the Injury Impairment Scale (IIS) were compared with reported disability for work or school, household activities, and activities of daily living during the first 18 months after injury for 2,194 persons treated in an emergency department or hospitalized for their injuries. Persons whose most severe injury was more life-threatening or who had larger numbers of injuries more often had higher IIS scores for their most life-threatening injury than did persons with fewer or less life-threatening injuries. Hospitalized patients had higher IIS injuries more often than did emergency department patients. As IIS increased, duration of disability also increased. However, many high IIS patients either reported no disability or disability of only short duration, suggesting only limited association between disability and IIS as it is currently designed. PMID- 7474015 TI - Treatment charges, payment sources, and disability from alcohol-related trauma. AB - We examined treatment charges and who paid them and disability for work, school, household activities, and activities of daily living according to identified use of alcohol before injury among 2,416 patients age 15 years or older from 22 northwestern Vermont communities. Over 90% received emergency department treatment only. Among patients reported to have consumed this drug mean hospital and physician charges, respectively, were $2,482 and $565; 31.7% did not pay any portion of their hospital bills, and 27.8% of their total bills were unpaid. Among patients not known to have consumed alcohol, mean hospital and physician charges were $601 and $158, respectively; 10.7% made no payments and 11.3% of their bills were unpaid. Bills of alcohol users more often were paid by Medicaid and less often by Workmen's Compensation or commercial insurance than were those of patients without alcohol. Users also were less likely than the nonalcohol group to pay physicians' bills for hospital care. Alcohol users experienced longer postinjury disability than did persons not known to have been drinking. PMID- 7474014 TI - Analysis of preventable trauma deaths and inappropriate trauma care in a rural state. AB - OBJECTIVE: The goal of this study was to determine the rate of preventable mortality and inappropriate care in cases of traumatic death occurring in a rural state. DESIGN: This is a retrospective case review. MATERIALS AND METHODS: Deaths attributed to mechanical trauma throughout the state and occurring between October 1, 1990 and September 30, 1991 were examined. All cases meeting inclusion criteria were reviewed by a multidisciplinary panel of physicians and nonphysicians representing the prehospital as well as hospital phases of care. Deaths were judged frankly preventable, possibly preventable, or nonpreventable. The care rendered in both preventable and nonpreventable cases was evaluated for appropriateness according to nationally accepted guidelines. MEASUREMENTS AND MAIN RESULTS: The overall preventable death rate was 13%. Among those patients treated at a hospital, the preventable death rate was 27%. The rate of inappropriate care was 33% overall and 60% in-hospital. The majority of inappropriate care occurred in the emergency department phase and was rendered by one or more members of the resuscitation team, including primary contact physicians and surgeons. Deficiencies were predominantly related to the management of the airway and chest injuries. CONCLUSIONS: The rural preventable death rate from trauma is not dissimilar to that found in urban areas before the implementation of a trauma care system. Inappropriate care rendered in the emergency department related to airway and chest injury management occurs at a high rate. This seems to be the major contributor to preventable trauma deaths in rural locations. Education of emergency department primary care providers in basic principles of stabilization and initial treatment may be the most cost effective method of reducing preventable deaths in the rural setting. PMID- 7474016 TI - Does the potential for organ donation justify scene flights for gunshot wounds to the head? AB - OBJECTIVE: The goal of this study was to evaluate helicopter transport to an urban level I trauma center from the scene of injury for patients with self inflicted gunshot wounds to the head. DESIGN: This study is a retrospective review of the prehospital, hospital, and billing records. MEASUREMENTS AND MAIN RESULTS: Despite the fact that 10 of 28 patients (36%) had an airway established by the medical flight crews, scene flights did not enhance survival. Twenty-seven of 28 patients (96%) died. The remaining patient's survival could not be attributed to the scene flight. We estimated that 27 of 28 patients would have arrived at the trauma center sooner if they had been transported by the first responder emergency medical services ground unit. Flight service charges were approximately one-third of the hospital charges. As a group, patients with a self inflicted gunshot wound to the head had the highest rate of organ donation in this trauma center (26%). Twenty-nine organs were harvested from the seven donors. CONCLUSIONS: The use of helicopter scene flights from the scene of injury for patients with a self-inflicted gunshot wound to the head provides no medical advantage to the victims, but provides a high-yield source of desperately needed organs. The prompt establishment of an airway in the field may prolong patient survival long enough to allow evaluation for organ donation. Helicopter transport of these patients is justified only as a means of rapidly delivering the personnel capable of providing advanced airway skills to the scene. Patients requiring CPR in the field after isolated gunshot wounds to the head will not live long enough to become organ donor candidates; therefore, there is no benefit to helicopter transport for these patients. PMID- 7474017 TI - The evaluation of a two-tier trauma response system at a major trauma center: is it cost effective and safe? AB - OBJECTIVE: To determine the efficacy and safety of a two-tier trauma response, using prehospital criteria for matching trauma center assets with severity of injury. DESIGN: A prospective iterative study on a consecutive sample of patients to test the hypothesis. MATERIAL AND METHODS: Criteria were developed whereby in hospital response was determined by information provided by prehospital personnel. Two modifications of these criteria were introduced at 6 and 9 months. Triage and response accuracy were evaluated using outcome variables. Cost savings were estimated using differences between the full and modified teams. Chi-squared analysis was used. MEASUREMENTS AND MAIN RESULTS: Of 1,479 patients evaluated over a 9-month period, 682 (46%) received a full trauma team response, and a modified trauma team responded to 794 (54%). When compared with final designation by outcome variables, the sensitivity, specificity, and accuracy were significantly improved after the first modification of criteria. After the second modification, there was no significant improvement; however, the number of undertriaged patients increased significantly. Estimated cost savings were about $178,000 over the 9-month period. CONCLUSIONS: Utilization of a two-tier response to trauma patients is effective, safe, and results in substantial cost savings. PMID- 7474018 TI - Esmarch closure of laparotomy incisions in unstable trauma patients. AB - Fascial closure after laparotomy may be time-consuming and extremely difficult, especially in the setting of massive bowel edema. In the trauma patient with deteriorating hemodynamic status, hypothermia, or worsening hypoxia, expeditious abdominal wall closure is essential to facilitate rapid transport to the intensive care unit for further stabilization. With the increasing utilization of the abbreviated laparotomy in unstable trauma patients, innovative techniques for speedy fascial closure must be evaluated. We developed the Esmarch closure--a simple, rapid method for closing the abdominal wall at the end of abbreviated laparotomies. PMID- 7474019 TI - Improvement in cost recovery at an urban level I trauma center. AB - With escalating health care costs and health reimbursement reorganization, the greatest danger to trauma centers will remain expensive uncompensated care. This is caused primarily by costs incurred in treating complex, life-threatening injuries and to the large population of trauma patients with no, or inadequate, means of compensation. In 1986 and 1987, this urban level I trauma center experienced an operating loss totaling $2,335,200. To attempt to reverse this expense, annual tracking of the trauma service's financial performance was begun in 1989. In addition, changes were made. Early multidisciplinary baseline assessment of each admission was instituted for financial profiling and discharge planning. Attempts were made on admission to identify health and vehicular insurance information. Processing for Medicaid and Medicare reimbursement was begun as soon as possible, and coding for diagnoses was checked by medical records personnel and the trauma nurse coordinator. If appropriate, Missouri Crime Victims Compensation was sought. Base on costs incurred in providing trauma services, as required by the state of Missouri and the American College of Surgeons, a trauma response charge was developed and instituted. Over a 5-year period, 1989 to 1993, financial audits were conducted. The cost recovery ratio (CRR) (collections/cost) was utilized as the measure of financial success. The CRR improved from 0.74 in 1989 to 0.93 in 1993, and in 2 years, 1991 and 1992, was 1.03 and 0.99. Over this period, the acuity of injury, as measured by the Injury Severity Score, remained essentially the same, but length of hospital stay decreased from 10.0 to 8.7 days. The CRR was greatest for private insurance.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7474020 TI - Permissive hypercapnia. AB - Traditional practice of mechanical ventilation includes tactics to reduce lung injury, such as avoidance of excessive airway pressure, patient distress, and tidal volume. Gas exchange objectives have received priority, however, and a degree of lung injury has been accepted as inevitable. The current trend toward increasing use of permissive hypercapnia is based on the recognition that lung injury induced by mechanical ventilation may be reduced by compensated hypercapnia with few serious adverse effects and contraindications. PMID- 7474021 TI - Dislocation of the shoulder and ipsilateral fracture of the humeral shaft: case reports and literature review. AB - Shoulder dislocation associated with ipsilateral humeral shaft fracture is an uncommon combination. Two cases treated, one by open and the other by closed methods, are presented. The mechanism of this complex injury is detailed and the treatment modalities are discussed. The relevant literature is reviewed. PMID- 7474022 TI - Posterior tibial tendon tear combined with a fracture of the accessory navicular: a new subclassification? AB - A 58-year-old woman with a diagnosis of a tear of the posterior tibial tendon associated with the os accessory navicular fracture was treated surgically. We believe that the dysfunction of the posterior tibial tendon was associated with an os accessory navicular fracture. This case is proposed as a new subclassification that was separate from the traditional classification of the tear of the posterior tibial tendon. PMID- 7474023 TI - Isolated avulsion fracture of the lesser tuberosity of the humerus. AB - Six cases of isolated avulsions of the lesser tuberosity of the humerus were encountered within a short period of time. Although it seldom has been described in the literature, it might be a more common fracture than suspected. Physical examination revealed a painful shoulder with limited function in all six patients. In retrospect, all diagnoses could have been made on the primary plain roentgenograms. However, the fragment of the lesser tuberosity was easily misdiagnosed. An axillary view showed the fragment clearly in all cases. Computed tomography is useful for making an accurate diagnosis because it establishes the size and displacement of the fragment and determines additional injuries, such as a ruptured biceps tendon. Five patients with displacement of the lesser tuberosity were operated on after several weeks or months. The lesser tuberosity was reattached in all cases. After 6 months three patients had slight impairment of elevation and external rotation, and one of them suffered from pain during movements of the arm. The patient with no fracture displacement was treated conservatively with a good result. In conclusion, we recommended an axillary roentgenogram as part of the posttrauma radiologic series of the shoulder. A nondisplaced avulsion fracture of the lesser tuberosity of the humerus can be treated conservatively. We recommend reattachment of displaced fractures. PMID- 7474024 TI - The 1994 distinguished lecturer in substance abuse. PMID- 7474025 TI - A psychoanalyst in methadonia. AB - This paper argues that the contemporary psychoanalytic model of addiction is compatible with the biochemical deficit model of Dole and Nyswander, the basis of methadone pharmacotherapy. The author explains the self-object function of methadone in the repair of self-deficits, specifically in the area of affect regulation, and he expands the dynamics of self-repair to the relational and programmatic "context of care" of methadone maintenance. Clinically, the problems of resistance and transference are examined as they arise and are resolved within an intersubjective counseling relationship. The treatment process in methadone maintenance is then reconceptualized as the gradual transformation of the central organizing principles of the addicted person as a result of the experience of psychophysiological stability, complemented by acceptance and empathy within a secure context of care. In closing, the need for an individual, self-focused approach to deciding between maintenance and detoxification is suggested. PMID- 7474026 TI - The development of the Comprehensive Addiction Severity Index for Adolescents (CASI-A). An interview for assessing multiple problems of adolescents. AB - The Comprehensive Addiction Severity Index for Adolescents (CASI-A) is a 45 to 90 minute comprehensive, semi-structured clinical interview for evaluating adolescents who present for treatment at various provider agencies. CASI-A modules and their individual items were selected and revised based on theory, clinical wisdom, and adolescent experiences obtained during pilot interviews and focus groups. The CASI-A assesses known risk factors, concomitant symptomatology, and consequences of adolescent alcohol/drug use within seven primary areas of functioning: education status, alcohol/drug use, family relationships, peer relationships, legal status, psychiatric distress, and use of free time. The CASI A is not a diagnostic or screening instrument, but rather a clinical assessment tool that obtains clinically pertinent information designed to guide treatment planning and to evaluate treatment outcome. The CASI-A's design makes it suitable for administration in a variety of settings, for repeat administration at posttreatment follow-up evaluations, and for assessment of virtually all adolescents in treatment regardless of their admission problem. Overall, the CASI A has encouraging but preliminary evidence of validity and internal consistency. Information collected soon after admission during administration of the CASI-A by nonclinical interviewers corresponded quite well with that obtained over the course of the adolescent's treatment stay by the entire treatment team. Revisions to the instrument are being made in those areas where correspondence between information on the CASI-A and that extracted from clinical records dropped below 75%, or in those early subscales, where alpha coefficients dropped below .6. As a result of the encouraging results reported in this paper, we are beginning additional psychometric testing, refining the proposed scoring system, and developing a computerized data entry, scoring, and report system. PMID- 7474028 TI - Effects of an appointment reminder call on patient show rates. AB - A pilot study (N = 80) was conducted to determine if (1) prospective substance dependent patients randomly selected to be reminded (TC) of their scheduled intake evaluation the day before their first appointment would have a higher show rate than those not contacted (NC); and (2) if TC subjects administered a satisfaction questionnaire 1-3 days after intake would exhibit higher treatment retention rates at one week and one month posttreatment entry than NC subjects not exposed to the questionnaire. The findings suggest that reminding prospective patients of their initial scheduled appointments and following up with phone calls to those who fail to show can improve the rate at which patients will initiate treatment, provided initial appointments are scheduled in a timely manner (7 days or less). Similarly, the combination of the reminder call and the satisfaction questionnaire were associated with higher treatment retention rates for those whose initial appointments were scheduled in a timely manner. PMID- 7474027 TI - Acupuncture for the treatment of cocaine addiction. Investigation of a needle puncture control. AB - We conducted a 6-week, single-blind study of acupuncture for cocaine dependence in methadone-maintained patients (N = 40) for the purpose of identifying an appropriate needle puncture control for use in future large-scale clinical trials. Patients were randomly assigned to receive daily acupuncture in three auricular sites plus one body site (LI-4), or in control sites within 2-3 mm of the four active sites. Overall, there was a positive response to treatment on a variety of drug-related and psychosocial measures. Cocaine use decreased significantly for patients in both needle puncture groups. The only statistically significant difference between the two types of needle puncture was on ratings of craving. Subjects rated each type of needle puncture as equally credible and perceived no significant differences on the acute effects of the two types of needle insertions. Power calculations based on these findings suggest that very large sample sizes would be required to detect treatment differences between this control condition and active acupuncture. Alternative controls are suggested, and the challenges inherent in implementing controlled clinical trials of acupuncture are discussed. PMID- 7474029 TI - Individual Assessment Profile (IAP). Standardizing the assessment of substance abusers. AB - The Individual Assessment Profile (IAP), a structured intake assessment interview instrument in the public domain, was designed for use with substance-abusing populations in several large-scale and community-based drug abuse treatment projects underway in the United States. Background information is presented, including the content-based item-selection process used during the pretests and pilot testing. Validity and test-retest reliability data are also presented, along with descriptions of studies using the IAP for clinical, research, and management information purposes. Concordance between biological measures and self reports of recent drug use, measures of internal consistency, and test-retest reliability coefficients were generally good. A computer-assisted personal interview version of the IAP and an automated reporting system were subsequently developed for clinical and management reporting purposes and used in a large scale research demonstration project. An intreatment version of the IAP has also been developed to collect information on treatment services provided and to assess changes in behaviors after 3, 6, and 12 months of treatment. These instruments (the IAP intake and intreatment interviews) provide a comprehensive system to assess substance-abusing populations. PMID- 7474030 TI - Program monitoring for clinical practice. Specimen positivity across urine collection methods. AB - Urinalysis is a primary component of methadone programming. A clinic-based study was conducted to determine whether unsupervised urine collection is as reliable as other methods for detecting unauthorized drug use among methadone treatment clients. Drug positivity rates compared over a 30-week period revealed no significant differences as a function of collection method. Results confirm the efficacy of unsupervised urine collection using a heat sensitive strip or temperature indicator. Study findings also offer evidence to support clinical practice designed to ensure client privacy and dignity while maintaining standards for effective program monitoring. PMID- 7474031 TI - Combining behavioral and self-help approaches in the inpatient management of dually diagnosed patients. AB - The combination of a token economy and a self-help model made it feasible to organize and maintain an intensive dual-diagnosis treatment program in a public municipal hospital acute psychiatric ward. The program described here, which integrates the token economy and 12-step approaches, fosters voluntary and active patient participation in the process of simultaneous recovery from mental illness and addiction; it also networks the patient with community resources. Implementation of the clinical program involved addressing multiple clinical and organizational obstacles, including a multihandicapped and often nonmotivated patient population, a lack of psychiatric staff trained in managing substance abuse, and the need to integrate mental health with substance abuse clinical interventions. The program has been operating more than 5 years and has treated more than 1,000 patients. As the token economy became an integral part of the ward routine, violence on the ward declined substantially, and patient participation in group activities increased significantly. PMID- 7474032 TI - The physiologic and pathologic roles of Tamm-Horsfall glycoprotein in urinary system. AB - Tamm-Horsfall glycoprotein (THG), a unique renal glycoprotein, is synthesized exclusively by the epithelial cells of the thick ascending loop of Henle and early segment of distal convoluted tubules. The protein is the primary constituent of urinary hyaline cast. The aggregation and gel formation of THG in response to increasing concentrations of electrolytes and low pH within physiologic ranges may affect the permeability of water in this segment. The dual effect of THG on mononuclear cells renders the molecule to become a potent immunomodulator. The cytokine-binding activity of THG lets the kidney an important tissue for cytokine catabolism. The aggregation characteristics of THG also plays a role in pathologic states and leads to the persistence of interstitial deposits in several tubulointerstitial diseases. The induction of immune responses to this protein, including urinary tract infection, reflux nephropathy chronic active hepatic damage, elicites immune complex tubulointerstitial nephritis. It is expected that the detection of THG or its antibodies may aid in the diagnosis of renal tubular disorders. PMID- 7474033 TI - Application of natural and amplification-created restriction sites in prenatal and preimplantation diagnosis of beta-thalassemia. AB - We attempted the strategy of natural and amplification-created restriction sites for early prenatal and preimplantation diagnosis of Chinese beta-thalassemia. Mutagenesis primers were designed for 11 mutations reported for the Chinese population. The diagnosis was established after polymerase chain reaction and digestion of products by specific enzymes. The results were confirmed by direct sequencing of enzymatically amplified double-stranded DNA. The beta-globin gene was amplified from triploid embryos and isolated blastomeres using mismatched primers. The mutant and normal alleles can be distinguished clearly by this new method. Early prenatal diagnosis was successfully achieved in 9 cases. The beta globin gene was successfully amplified from single blastomeres and tripronuclear embryos with mismatched primers. Natural and amplification-created restriction sites are a reliable method for rapid prenatal diagnosis of beta-thalassemia. Furthermore, the strategy provides a possible approach for the preimplantation diagnosis of beta-thalassemia. PMID- 7474034 TI - A molecular study of ocular herpes simplex infection in Kaohsiung, Taiwan. Part I: Cleavage pattern analysis. AB - In order to investigate the relationship between genotypes of herpes simplex virus (HSV) and its clinical pictures in herpes simplex virus keratoconjunctivitis (HSK), fifty nine strains of HSV isolates, collected in the Kaohsiung Medical College Hospital during January 1981 to July 1991, and F strain (ATCC VR-73), a standard HSV-1, were studied. The viruses were isolated from HSK patients. All 59 strains were identified as HSV-1 using immunofluorescence monoclonal antibody method. The HSV DNAs were extracted and digested by restriction enzymes, BamH I, Kpn I and Sal I, followed by electrophoresis, and photographing. Six kinds of DNA cleavage patterns were found in Taiwan. The majority were E subtype (56.4%) then A subtype (27.3%), D subtype (7.3%), F subtype (5.4%), G subtype (1.8%) and H subtype (1.8%). Subtypes B and C were not found in this study. The cleavage patterns of two recurrent cases belonged to E and A subtypes. The identical cleavage patterns of two bilateral infections all belonged to the E subtype. PMID- 7474035 TI - Inhibitory effect of ethanol on voltage-dependent potassium currents in single aortic smooth muscle cells. AB - The effect of ethanol on voltage-dependent K+ currents was examined in single aortic smooth muscle cells. The aortic smooth muscle cells were isolated from guinea pigs. The tight-seal patch clamp technique in whole cell configuration was employed. Ethanol (500 mM) reversibly inhibited voltage-dependent transient outward K+ current, the spontaneous transient oscillatory K+ current (STOC) and steady-state K+ current, whereas ethanol (50 mM) did not produce any change in these K+ currents. When the peak-height amplitudes of STOC were analyzed, the application of ethanol (50 mM) was noted to produce significant inhibition on both amplitude and frequency of STOC evoked at the holding potential of +10 mV. The results shown in the present study indicate that ethanol-induced inhibition of the voltage-dependent K+ channel, which is not required for the elicitation of either GTP-binding protein(s) or inositol triphosphate, may contribute to its vasoconstrictor effect because the inhibition of the voltage-dependent K+ channel may lead to the increase of cell excitability in vascular smooth muscle. PMID- 7474036 TI - The relationship of alveolar permeability and pulmonary inflammation in patients with systemic lupus erythematosus. AB - The alveolar epithelium permeability (AP) in 34 patients with systemic lupus erythematosus (SLE) was measured by Tc-99m DTPA radioaerosol inhalation lung scintigraphy. The degree of AP damage in SLE was represented as the slope of the time-activity curve from the dynamic lung imagings. The patients were divided into two subgroups according to the following two criteria. [A] clinical feature: they were separated as stable or flare stage; and [B] chest X-ray findings: they were separated as positive or negative results. Meanwhile, the quantitative Gallium-67 (Ga) lung scan was performed as Ga uptake index (GUI) to evaluate the severity of inflammation in the lungs in SLE. The results show that [1] there were no significant statistical differences in the degree of AP damage between the two subgroups according to clinical features or chest X-ray findings, and [2] no good correlation between the degree of AP damage and inflammation of the lungs was found. In conclusion, the change of AP in SLE may be a mechanism not related to the presentation of clinical features or chest X-ray findings, and the degree of AP damage in SLE is not related to the severity of pulmonary inflammation. PMID- 7474037 TI - Hepatocellular carcinoma with metastasis to right atrium--a report of three cases. AB - Hepatocellular carcinoma (HCC) is one of the most common neoplasma in Taiwan. The tumor itself has the tendency of extension into the venous system, such as to the portal vein, hepatic vein and inferior vena cava (IVC), but intra-atrial metastasis is unusual. Antemortem diagnosis was difficult before the availability of two-dimensional echocardiography (2-DE). Sometimes, the first symptoms and signs are cardiogenic manifestations such as dyspnea on exertion, syncope, edema of the lower legs, and shock. Clinicians may mistakenly make the wrong diagnosis of heart failure. Because of this, we hereby report three cases of HCC with right intra-atrial metastasis to raise the physician's awareness. All three cases initially presented as right side heart failure. Imaging study revealed hepatocellular carcinoma with right intra-atrial metastasis. Two of the three cases died within one month after diagnosis. PMID- 7474038 TI - [The incidence and mortality rates of head injuries in Kaohsiung City, Taiwan (1991-1992)]. AB - Head injury is a leading cause of mortality, morbidity and disability in Taiwan. This study was based on data from the investigation of head injuries in Kaohsiung City, Taiwan, during the period July 1, 1991 to June 30, 1992. The 3729 cases identified represented an overall incidence rate of 267/100,000--359/100,000 for males and 170/100,000 for females. The highest incidence rate was seen in the elderly group, followed by the 20-29 year age group. The population with the highest risk was males aged 20-29 years, with an incidence rate of 585/100,000. Seventy percent of all cases were from motor vehicle-related causes, followed by falls (15.3%) and assaults (8.7%). Of the motor vehicle-related head injuries, 70.9% resulted from motorcycle accidents. The incidence rate of motor vehicle head injuries was 188/100,000--248/100,000 for males and 125/100,000 for females. At all ages motor vehicle head injuries were higher in males than in females. Mortality rates from head injuries were 26/100,000--38/100,000 for males and 14/100,000 for females. The overall case fatality rate was 10%, and 61% of all deaths occurred prior to hospitalization. The case fatality rate was highest in older age groups. Initial clinical assessment was recorded using the Glasgow Coma Scale. Eighty-one percent of the cases were considered mild, and 19% moderate to severe in degree including 367 deaths. The males and the aged not only had a higher incidence of head injuries but also had a higher proportion of more severe ones. These findings indicate that head injuries are nonrandom and likely to occur in certain types of individuals under certain circumstances.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7474039 TI - [Secretory carcinoma of the breast--a case report]. AB - Secretory breast carcinoma is a very rare tumor of the breast and has distinctive pathological characteristics such as (1) the presence of large amounts of intracellular and extracellular secretion and (2) granular eosinophilic cytoplasma of the cells. It usually occurs in children or adolescents and is known to have a better prognosis than the usual ductal carcinoma. The preoperative aspiration cytology is also distinctive and it could permit planning the optimal surgical therapy such as breast conservation surgery for young patients. We report a case of secretory breast carcinoma in a 35 year-old female and her preoperative aspiration cytology in Liu's stain. To our knowledge, Secretory breast carcinoma has not been reported previously in Taiwan. PMID- 7474040 TI - Influence of a 36-h fast on the central and peripheral components of VO2 during submaximal exercise and peak oxygen uptake. AB - To determine if a 36-h fast affects the central and peripheral components of VO2 during submaximal exercise and peak oxygen uptake (VO2 peak), 10 physically healthy males exercised first for 10 min at approximately 60% of their VO2 peak and then to fatigue. Their cardiovascular responses were compared with those of 10 physically healthy controls (males) who were individually matched by VO2 peak values. Fasting had no significant effect on the central (heart rate, HR; stroke volume, SV; cardiac output, Q) and peripheral (arteriovenous oxygen content difference, a-vO2 diff.) components of VO2 during submaximal exercise. The respiratory exchange ratio (RER) during submaximal exercise was significantly decreased in the fasted subjects. Based on the RER, fasting altered substrate utilization of the muscle and thereby increased the contribution of fat during submaximal exercise. Fasting had no significant effect on VO2 peak, HR or O2 pulse, but RER was significantly lower during incremental exercise to fatigue. These results demonstrate that VO2 during submaximal exercise in fasted subjects is achieved by similar central and peripheral responses as found in non-fasted subjects, and that fasting does not reduce VO2 peak. PMID- 7474041 TI - Influence of ingesting a carbohydrate-electrolyte solution on endurance capacity during intermittent, high-intensity shuttle running. AB - The aim of this study was to examine the effects of ingesting a carbohydrate electrolyte solution on endurance capacity during a prolonged intermittent, high intensity shuttle running test (PIHSRT). Nine trained male games players performed two exercise trials, 7 days apart. On each occasion, they completed 75 min exercise, comprising of five 15-min periods of intermittent running, consisting of sprinting, interspersed with periods of jogging and walking (Part A), followed by intermittent running to fatigue (Part B). The subjects were randomly allocated either a 6.9% carbohydrate-electrolyte solution (CHO) or a non carbohydrate placebo (CON) immediately prior to exercise (5 ml kg-1 body mass) and every 15 min thereafter (2 ml kg-1 body mass). Venous blood samples were obtained at rest, during and after each PIHSRT for the determination of glucose, lactate, plasma free fatty acid, glycerol, ammonia, and serum insulin and electrolyte concentrations. During Part B, the subjects were able to continue running longer when fed CHO (CHO = 8.9 +/- 1.5 min vs CON = 6.7 +/- 1.0 min; P < 0.05) (mean +/- S.E.M.). These results show that drinking a carbohydrate electrolyte solution improves endurance running capacity during prolonged intermittent exercise. PMID- 7474042 TI - The influence of strength training, swim training and ageing on the Achilles tendon and m. soleus of the rat. AB - Different age groups of male Wistar rates were submitted to intense strength training, swim training or no training. It was found that the tetanic of the m. soleus decreased with age. This was counteracted by strength training, whereas swim training had no effect on tetanic tension. Force at ultimate failure and yield point in the Achilles tendon decreased with age, but were not influenced by strength training. Swim training, on the other hand, appeared to compensate for the ageing process in the tendon. We conclude that tendon strength decreases with age and that tendon strength does not reflect muscle strength. The absolute tensile strength of tendons appears to be influenced by physical activity of endurance-type exercise. However, it is not known whether the tissue responds to the number of mechanical muscle contractions or physiological factors connected with endurance training. Although it is questionable to extrapolate from the rat to humans, it is speculated that intensive muscle strength training should be accompanied by endurance exercise in order to prevent the muscles from damaging connective tissue structures, such as the tendons and ligaments. PMID- 7474043 TI - Recreational exercise participation and aerobic fitness in men and women: analysis of data from a national survey. AB - We compared the relationship between predicted VO2 max (ml kg-1min-1) and recreational exercise patterns, using secondary data analysis of a comprehensive national survey (18,293 subjects aged 15-69 years). Exercise participation and predicted VO2 max data were available for about 50% of this sample (4933 females, 4738 males). As expected, VO2 max was significantly lower in the females than in the males at any age (P < 0.0001). Age was the most significant predictor of VO2 max (r = -0.71 for males, r = -0.73 for females). Adjusting the data for the body mass index (BMI) increased this relationship only slightly in the males (R = 0.75) and females (R = 0.79). The simultaneous inclusion of exercise participation data (intensity, duration, energy expenditure) did not increase the predictions meaningfully (R = 0.78 for the males, R = 0.81 for the females). These exercise participation parameters concomitantly accounted for only a very slight amount of the variance of VO2 max in both the females (3.0%) and males (4.5%). To minimize the effects of age, the data were analysed using 5-year intervals. Again, the exercise participation parameters accounted for only a small part of the variance in VO2 max (< 10%), except in the 15- to 19-year-old males (24%). These data suggest that VO2 max is not associated with participation in recreational exercise. PMID- 7474044 TI - Influence of aerobic versus anaerobic exercise on the relationship between reproductive hormones in men. AB - This study examined the effects of equal anaerobic and aerobic total work outputs on the relationship between reproductive hormones in men. Nine subjects performed three randomized trials on separate days: (1) 1 h period of rest (control), (2) 1 h continuous aerobic exercise (65% VO2 max), and (3) 1 h intermittent anaerobic exercise (which included 2 min exercise periods at 110% VO2 max). The total work output of the aerobic and anaerobic trials were equated. For the 8 h after each experimental trial, blood samples were collected hourly and analysed for testosterone, luteinizing hormone (LH), follicle-stimulating hormone (FSH), prolactin and cortisol. Diet, physical activity and circadian influences were all controlled. Compared with the control, the aerobic and anaerobic trials significantly (P < 0.05) elevated testosterone, prolactin and cortisol; however, these changes were transient and returned to control levels within 1-2 h of recovery. Neither exercise produced significant (P > 0.05) changes from control for LH and FSH. The area under the hormonal response curves (AUC) was calculated for the 8-h recovery period. The testosterone and LH AUC results did not differ (P > 0.05) among the trials (the FSH AUC was not calculated). The prolactin AUC for the aerobic and anaerobic trials were greater (P < 0.01) than the control trial. The cortisol AUC for the anaerobic trial was greater than both the control and aerobic trials (P < 0.05), but the aerobic and control trials did not differ from one another.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7474045 TI - Shooting performance is related to forearm temperature and hand tremor size. AB - The changes in postural tremor of the hand and the subsequent effect on shooting performance produced by moderate cooling and heating of the forearm were studied in six subjects. Cooling produced a large decrease in tremor size of the ipsilateral hand, whereas warming the limb produced an increase in tremor size. Cooling or warming the forearm did not change the peak frequency of tremor significantly, which was quite stable for each subject. The improvement in shooting performance after cooling the forearm, as measured by grouping pattern of the shots, reached statistical significance and warming caused a significant worsening. This measure of performance was shown to correlate (r = 0.776) inversely with tremor size. The causes and implications of these changes are discussed. It is suggested that local cooling may be useful for people who wish temporarily to reduce tremor in order to improve dexterity for shooting and for other purposes. PMID- 7474046 TI - Determination of physiological data from a mathematical analysis of the running performance of elite female athletes. AB - A method of analysis developed and applied to male athletes is used to determine physiological data for female athletes. The analytical method rests on the construction of an energy balance incorporating a mathematical representation of each of the major terms in the balance. By recasting the energy balance, a relationship between race distance and running time is derived. Excellent agreement with results for female athletes from recent Olympic Games for distances from 100 to 10,000 m is established. From the analysis, representative values of the maximum anaerobic power, the maximum aerobic power and the rate of degradation of mechanical energy into heat are derived for elite female athletes. PMID- 7474047 TI - The effect of a 12-over spell on fast bowling technique in cricket. AB - Nine members of the Western Australian Cricket Association fast bowling development squad were selected to determine the effects that a 12-over spell would have on fast bowling technique and selected physiological variables. Three high-speed cameras operating at 100 Hz filmed the subjects bowling the 5th and 6th balls of their 1st, 6th, 10th and 12th overs. Blood lactate and heart rate were recorded and the bowlers were shown to work at between 80.3% (1st over) to 84.7% (12th over) of their maximum heart rate during the 12-over bowling spell. Data from the 5th and 6th deliveries from each over were averaged to provide representative data, as no significant differences were evident between these two deliveries for the selected kinematic variables. For all subjects, no significant differences were recorded for the selected kinematic variables throughout the duration of the 12-over spell, indicating that in general fast bowling technique does not change over this length of spell. In addition, there was some evidence of change in technique for the bowlers who used a front-on action, where counter rotation of the shoulders was found to increase; however, this could only be confirmed by a more comprehensive study. PMID- 7474048 TI - The augmenting role of biofeedback: effects of autogenic, imagery and music training on physiological indices and athletic performance. AB - In this study, three psychoregulative procedures of relaxation and excitation were provided in combination with biofeedback to examine their role on physiological and athletic performance variables. Thirty-nine college students were randomly assigned to three groups of psychoregulatory treatment (autogenic and imagery training, AT+IT; music and imagery training, M+IT; autogenic, music and imagery training, AT+M+IT), one placebo group and the control group. Imagery was related to a 100-m run. The treatment and control conditions lasted 13 sessions of 20 min each. During the first seven sessions, the subjects in the treatment groups underwent 10 min of relaxation followed by 10 min of excitation. During the last six sessions, similar treatment was provided accompanied by frontalis EMG biofeedback. Heart rate, the galvanic skin response, EMG and breathing frequency (fb) were recorded three times during each session. In addition, an athletic task (100-m run) was examined at the outset, after seven sessions (no biofeedback) and after an additional six sessions (with biofeedback). Biofeedback was found to have a significant augmenting effect on physiological components and athletic performance when accompanied by autogenic, imagery and music training. Soft music was found to be as beneficial as other relaxation techniques. The results are compared with similar studies applying mental techniques with biofeedback, and new directions of investigation in the psychophysiological domain are suggested. PMID- 7474050 TI - Advances in ultrasound technology. PMID- 7474049 TI - Calibrating skinfold calipers. AB - While athletes are routinely assessed for changes in subcutaneous adipose tissue with skinfold calipers, absolute dynamic calibration of caliper jaw compression is currently not possible. The first part of this study describes how dynamic compression of foam rubber blocks can be used to monitor the relative calibration of a single pair of calipers as springs fatigue, or to alert an investigator to variations in measurement values between different calipers. The second part of the study, carried out on 10 female athletes, demonstrated that the significant differences established by the foam block method of calibration also translated into a significant difference for the sum of seven subcutaneous skinfolds. Foam blocks can be used as a simple, inexpensive method to establish a calibration range and can also be used to recheck calipers periodically, depending upon their use. PMID- 7474051 TI - Ultrasonography in the detection of aneuploidy in the first trimester. AB - Ultrasonography has been used increasingly in the first trimester to identify fetal anomalies as early as possible in gestation. First trimester sonographic screening may detect a greater proportion of aneuploid fetuses than sonography performed later in pregnancy, and it may allow determination of potential markers for prediction of chromosomal anomalies. PMID- 7474052 TI - Nuchal translucency thickness and crown-rump length in twin pregnancies with chromosomally abnormal fetuses. AB - We retrospectively examined the crown-rump length and nuchal translucency thickness of each fetus in eight twin pregnancies in which karyotyping at 10 to 14 weeks' gestation demonstrated that at least one of the fetuses was chromosomally abnormal. Eight fetuses had trisomy 21 and two had trisomy 18. The nuchal translucency thickness was more than 2.5 mm in nine (90%) of the trisomic fetuses and in one of the chromosomally normal ones. In contrast, the crown-rump length was below the fifth percentile in only one of the fetuses with trisomy 18; all other measurements were within the normal range. PMID- 7474053 TI - Fetal exomphalos at 11 to 14 weeks of gestation. AB - In an ultrasonographic screening study at 11 to 14 weeks' gestation involving 9885 singleton pregnancies, the prevalence of exomphalos was 0.11% (11 cases) and the prevalence of trisomy 18 or 13 was 0.35% (35 cases). The mean maternal age of the screened population was 35 years (range, 15 to 47 years) and a significant association was found between maternal age and both the prevalence of trisomies and the prevalence of exomphalos. Because the frequency of exomphalos in fetuses with trisomy 18 or 13 was 17% and in those with no evidence of these trisomies it was 0.05%, the risk for trisomies in fetuses with exomphalos is 340 times higher than in those without exomphalos. PMID- 7474054 TI - Pilot screening for fetal malformations: possibilities and limits of transvaginal sonography. AB - Two thousand and ninety-seven unselected pregnant women bearing 2114 fetuses were examined by transvaginal sonography at 14 weeks of gestational age and rescreened via transabdominal sonography at 21 weeks (excluding those who chose termination of pregnancy). Twenty-five of 47 anomalies were correctly identified at the first scan, 15 malformations were missed and detected only during the scan, two were identified later in pregnancy, and five were identified after birth. Ten of 12 abnormalities were detected correctly on the basis of ultrasonographic findings. Transvaginal sonographic screening for fetal malformation, in our experience, permits the detection of more than 50% of all fetal structural defects and 75% of all aneuploidies early in pregnancy. PMID- 7474055 TI - Early prenatal diagnosis of axillary cystic hygroma. AB - Fetal axillary cystic hygroma has been reported rarely and only as a sonographic finding late in gestation. A retrospective study of 19,200 early second trimester screening ultrasonograms for anomalies revealed five cases of axillary cystic hygroma. All of the axillary hygromas were small, transient, nonloculated cysts. Three cysts were associated with chromosomal abnormalities: trisomy 21 in two cases and a single case of trisomy 18 in which structural abnormalities also were detected. Early fetal axillary cystic hygroma appears to be different from postnatal axillary cysts on the basis of differences in prevalence, structure, and appearance. It also may be a normal variant in the development of the fetal lymphatic system. PMID- 7474056 TI - Early sonographically guided amniocenteses with filtration technique: follow-up on 249 procedures. AB - Fetal and neonatal outcome is reported for 249 consecutive low-risk pregnancies in which early amniocentesis was carried out with filter technique, at a mean gestational age of 12.5 weeks, to improve the yield in cell cultures. Three pregnancies (1.2%) were terminated because of abnormal test results, four stillbirths (1.6%) occurred between weeks 33 and 38, and five pregnancies were lost spontaneously after sampling, corresponding to 2% of unintended fetal losses before 28 weeks of gestation. Of the live-born infants, 2.9% were delivered prematurely, all between weeks 32 and 37. Malformations and orthopedic postural deformities were not associated with leakage after amniocentesis and were found at expected rates. None of the cases was lost to follow-up. The increase in abortion rate caused by early amniocentesis with filter technique is likely to be within the same range as that seen after routine amniocentesis and transabdominal chorionic villus sampling, although the sample is too small for a proper risk evaluation. PMID- 7474057 TI - Transvaginal sonographic diagnosis of fetal finger abnormalities in early gestation. AB - The prenatal sonographic diagnosis of fetal finger abnormalities was recorded in a study population of 20,000 pregnant patients. All scans were performed with a vaginal transducer at 13 to 17 weeks' gestation. The following abnormalities were observed: six cases of polydactyly, three of syndactyly, nine cases of overlapping fingers, two cases of cleft hands, two cases of adactyly, one case of aphalangia, and one case of clasped thumb. In 15 of the 25 fetuses with finger abnormalities, we observed associated malformations or chromosomal disorders, or both. We therefore recommend that examination of the fingers and hands should become an integral part of the early ultrasonographic evaluation of the fetus. PMID- 7474058 TI - Amniotic fluid accumulation in the first trimester. AB - Amniotic fluid volume was estimated sonographically in 507 of 977 first trimester pregnancies with living fetuses and in 99 of 227 instances of missed abortion. Volume is correlated with fetal length, cross referenced by average heart rate for living fetuses. Fluid volume increases exponentially after 9 weeks, consistent with the onset of renal function. Fluid volume tends to be excessive with missed abortion. Free fluid volume is a physiologic variable, determined from morphologic features. Standards are proposed for amniotic fluid volume dynamics in the first trimester. PMID- 7474059 TI - First and early second trimester diagnosis of anomalies of the central nervous system. AB - To study central nervous system development, 18 normally developing embryos or fetuses were examined sonographically between 6 and 16 weeks of pregnancy. Furthermore, in a 5 year period, 693 fetuses with an elevated risk for congenital defects were examined before 16 weeks of gestation. Abnormal development of the central nervous system was found in 28 fetuses. Early in pregnancy, normal central nervous system formation can be recognized, and pathologic cerebral development can be diagnosed already in the first trimester of pregnancy. We recommend routine use of safe and noninvasive transvaginal ultrasonography early in pregnancy for patients at risk. PMID- 7474060 TI - Early diagnosis of conjoined twins in triplet pregnancy after in vitro fertilization and assisted hatching. PMID- 7474061 TI - Congenital cytomegalovirus infection with oligohydramnios and echogenic bowel at 14 weeks' gestation. PMID- 7474062 TI - First trimester diagnosis of holoprosencephaly with a Dandy-Walker malformation by transvaginal ultrasonography. PMID- 7474063 TI - Fetal tricuspid valve dysplasia and pulmonary atresia at 14 weeks. PMID- 7474064 TI - Early prenatal diagnosis of Larsen's syndrome by transvaginal sonography. PMID- 7474065 TI - First trimester diagnosis of cyclopia and holoprosencephaly. PMID- 7474066 TI - Detection of twin-twin transfusion syndrome by first trimester ultrasonography. PMID- 7474067 TI - Natural history of antenatal omphalomesenteric duct cyst. PMID- 7474068 TI - First trimester transvaginal sonographic diagnosis of body stalk anomaly [correction of anatomy]. PMID- 7474069 TI - Cross-clade neutralization of primary isolates of human immunodeficiency virus type 1 by human monoclonal antibodies and tetrameric CD4-IgG. AB - We have tested three human monoclonal antibodies (MAbs) IgG1b12, 2G12, and 2F5) to the envelope glycoproteins of human immunodeficiency virus type 1 (HIV-1), and a tetrameric CD4-IgG molecule (CD4-IgG2), for the ability to neutralize primary HIV-1 isolates from the genetic clades A through F and from group O. Each of the reagents broadly and potently neutralized B-clade isolates. The 2F5 MAb and the CD4-IgG2 molecule also neutralized strains from outside the B clade, with the same breadth and potency that they showed against B-clade strains. The other two MAbs were able to neutralize a significant proportion of strains from outside the B clade, although there was a reduction in their efficacy compared with their activity against B-clade isolates. Neutralization of isolates by 2F5 correlated with their possession of the LDKW motif in a segment of gp41 near the membrane spanning domain. The other two MAbs and CD4-IgG2 recognize discontinuous binding sites on gp120, and so no comparison between genetic sequence and virus neutralization was possible. Our data show that a vaccine based on the induction of humoral immunity that is broadly active across the genetic clades is not impossible if immunogens that express the epitopes for MAbs such as 2F5, 2G12, and IgG1b12 in immunogenic configurations can be created. Furthermore, if the three MAbs and CD4-IgG2 produce clinical benefit in immunotherapeutic trials in the United States or Europe, they may also do so elsewhere in the world. PMID- 7474070 TI - A nonproducer, interfering human immunodeficiency virus (HIV) type 1 provirus can be transduced through a murine leukemia virus-based retroviral vector: recovery of an anti-HIV mouse/human pseudotype retrovirus. AB - The expression of a human immunodeficiency virus (HIV) type 1 provirus (F12-HIV) cloned from a nonproducer, chronically infected CD4 down-regulated Hut-78 cell clone (F12) does not lead to the formation of viral particles and, upon transfection in HeLa CD4+ cells, confers resistance to HIV superinfection without affecting the CD4 receptor exposure. In an attempt to transfer the anti-HIV properties of F12-HIV into human primary cell, we constructed a Moloney murine leukemia virus-based retroviral vector containing an F12-HIV genome lacking the 3' long terminal repeat and part of the nef gene, which was expressed under the control of its 5' long terminal repeat. The F12-HIV genome was inserted in the orientation opposite to that of the murine leukemia virus transcriptional unit and was designated the N2/F12-HIV nef-antisense vector. Lymphoblastoid CEMss cells, as well as human peripheral blood lymphocytes, were successfully transduced by the recombinant retrovirus emerging from the producer PA317 clones. CEMss clones expressing the F12-HIV nef-antisense vector became resistant to HIV superinfection even at the highest utilized multiplicity of infection (10(5) 50% tissue culture infective doses per 10(6) cells). In transduced CEMss cells the viral interference induced by the F12-HIV expression is not due to CD4 HIV receptor down-regulation. Nonproducer, interfering HIV proviruses transduced into retroviral vectors may, therefore, provide an alternative strategy for the protection of CD4+ human primary cells from HIV infection, which strategy may be used in designating a safe and efficient gene therapy protocol for patients with AIDS. PMID- 7474071 TI - Constitutive episomal expression of polypeptide IX (pIX) in a 293-based cell line complements the deficiency of pIX mutant adenovirus type 5. AB - The human adenovirus type 5 capsid is composed of a number of distinct polypeptides. It has been shown previously that one of these, polypeptide IX (pIX), is not absolutely required for the production of viable virus. However, viruses lacking this polypeptide have a significantly reduced packaging limit and, in the one case studied, also show a thermolabile virion phenotype. This report describes the use of eukaryotic episomal vectors based on the Epstein-Barr virus replicon to generate cells which stably express pIX. These cells provide pIX that is efficiently incorporated into virions that are genetically pIX-; such enhanced thermostability. These cells have also been used to isolate a genetically pIX- virus having a genome of length some 2.3 kbp in excess of the previously defined packaging limit for pIX- virus; the resulting virions have wild-type thermostability. These cells expand the theoretical capacity of adenovirus vectors for foreign DNA to around 9.2 kbp and may therefore be useful in gene therapy applications in which vector capacity is limiting. PMID- 7474072 TI - Interactions of the transcription factor AP-1 with the long terminal repeat of different human immunodeficiency virus type 1 strains in Jurkat, glial, and neuronal cells. AB - Human immunodeficiency virus type 1 (HIV-1) infection of the neuronal and astroglial cells of the central nervous system has been proposed to contribute to HIV-1-associated dementia. Recently it was shown that differences in the nucleotide sequence of the long terminal repeat (LTR) of different HIV-1 strains govern the tissue-specific pattern of viral expression. The LTR from central nervous system-derived HIV-1 strains JR-FL and JR-CSF directs expression in the neurons of transgenic mice, in contrast with the lymphotropic LAI strain. By in vitro footprinting, gel retardation, and methylation interference experiments, we have studied the interactions of host cell proteins from human neuronal, glial, HeLa, and Jurkat T cells with the LTRs from the neurotropic JR-FL and JR-CSF strains, compared with the LAI strain. Proteins belonging to the nuclear receptor family bind with different affinities to variant -352 to -324 sites. Gel supershift assays with Jun and Fos antibodies showed that the AP-1 transcription factor present in the various cell types was unable to recognize the -352 to -324 and -306 to -285 AP-1 putative binding sites. Interestingly, Jun and Fos components of AP-1 interact with the variant TGGCTCA sequence located in the -247 to -222 region of both neurotropic strains. These interactions were cell type specific, since they were detected only with extracts from glial and HeLa cells and not from neuronal or Jurkat cells. Cotransfection experiments further revealed that the -247 to -222 sequence is able to mediate AP-1-induced transcriptional activation in glial and not neuronal cells. PMID- 7474073 TI - Studies of the membrane fusion activities of fusion peptide mutants of influenza virus hemagglutinin. AB - Influenza virus hemagglutinin (HA) fuses membranes at endosomal pH by a process which involves extrusion of the NH2-terminal region of HA2, the fusion peptide, from its buried location in the native trimer. We have examined the amino acid sequence requirements for a functional fusion peptide by determining the fusion capacities of site-specific mutant HAs expressed by using vaccinia virus recombinants and of synthetic peptide analogs of the mutant fusion peptides. The results indicate that for efficient fusion, alanine can to some extent substitute for the NH2-terminal glycine of the wild-type fusion peptide but that serine, histidine, leucine, isoleucine, or phenylalanine cannot. In addition, mutants containing shorter fusion peptides as a result of single amino acid deletions are inactive, as is a mutant containing an alanine instead of a glycine at HA2 residue 8. Substitution of the glycine at HA2 residue 4 with an alanine increases the pH of fusion, and valine-for-glutamate substitutions at HA2 residues 11 and 15 are without effect. We confirm previous reports on the need for specific HAo cleavage to generate functional HAs, and we show that both inappropriately cleaved HA and mutant HAs, irrespective of their fusion capacities, upon incubation at low pH undergo the structural transition required for fusion. PMID- 7474074 TI - Functions of the internal pre-S domain of the large surface protein in hepatitis B virus particle morphogenesis. AB - The large hepatitis B virus (HBV) surface protein (L) forms two isomers which display their N-terminal pre-S domain at the internal and external side of the viral envelope, respectively. The external pre-S domain has been implicated in binding to a virus receptor. To investigate functions of the internal pre-S domain, a secretion signal sequence was fused to the N terminus of L (sigL), causing exclusive expression of external pre-S domains. A fusion construct with a nonfunctional signal (s25L), which corresponds in its primary sequence to sigL cleaved by signal peptidase, was used as a control. SigL was N glycosylated in transfected COS cells at both potential sites in pre-S in contrast to s25L or wild-type L, confirming the expected transmembrane topologies of sigL and s25L. Phenotypic characterization revealed the following points. (i) SigL lost the inhibitory effect of L or s25L on secretion of subviral hepatitis B surface antigen particles, suggesting that the retention signal mapped to the N terminus of L is recognized in the cytosol and not in the lumen of the endoplasmic reticulum. (ii) SigL was secreted into the culture medium even in the absence of the major HBV surface protein (S), while release of an L mutant lacking the retention signal was still dependent on S coexpression. (iii) s25L but not sigL could complement an L-negative HBV genome defective for virion secretion in cotransfections. This suggests that the cytosolic pre-S domain, like a matrix protein, is involved in the interaction of the viral envelope with preformed cytosolic nucleocapsids during virion assembly. PMID- 7474075 TI - Role of T-lymphocyte subsets in recovery from respiratory syncytial virus infection in calves. AB - The role of T-cell subsets in respiratory syncytial virus (RSV) infection was investigated by using monoclonal antibodies (MAbs) to selectively deplete gnotobiotic calves of CD4+, CD8+, or WC1+ gamma delta T-cell receptor+ lymphocytes. Injection of these MAbs produced specific reductions of the target cell populations in the circulation and tissues. Ten days after RSV infection, immunoglobulin M (IgM), IgG1, and IgA antibodies were detected in sera and lung washings from control calves. Depletion of CD8+ T cells had no effect on either the serum or local antibody responses to RSV, whereas depletion of CD4+ T cells suppressed the antibody responses in two of three calves. The IgM and IgA responses were significantly increased in the lung washings of calves from which WC1+ T cells were depleted. Depletion of CD4+ or WC1+ T cells caused no significant delay in virus clearance, although an increase in the extent of pneumonic consolidation was observed in anti-CD4-treated calves. Nasopharyngeal excretion of RSV was prolonged in calves depleted of CD8+ T cells, and virus was isolated in high titers from lung washings of these animals 10 days after infection, whereas virus had been cleared from lung washings of all other animals. The delayed virus clearance was associated with an increase in the severity of pneumonic consolidation in three of four of the calves from which CD8+ T cells were depleted. This study shows that CD8+ T cells play a dominant role in the recovery of calves from RSV infection. PMID- 7474076 TI - Hierarchy among multiple H-2b-restricted cytotoxic T-lymphocyte epitopes within simian virus 40 T antigen. AB - Simian virus 40 large tumor (T) antigen contains three H-2Db-restricted (I, II/III, and V) and one H-2Kb-restricted (IV) cytotoxic T lymphocyte (CTL) epitopes. We demonstrate that a hierarchy exists among these CTL epitopes, since vigorous CTL responses against epitopes I, II/III, and IV are detected following immunization of H-2b mice with syngeneic, T-antigen-expressing cells. By contrast, a weak CTL response against the H-2Db-restricted epitope V was detected only following immunization of H-2b mice with epitope loss variant B6/K-3,1,4 cells, which have lost expression of CTL epitopes I, II/III, and IV. Limiting dilution analysis confirmed that the lack of epitope V-specific CTL activity in bulk culture splenocytes correlated with inefficient expansion and priming of epitope V-specific CTL precursors in vivo. We examined whether defined genetic alterations of T antigen might improve processing and presentation of epitope V to the epitope V-specific CTL clone Y-5 in vitro and/or overcome the recessive nature of epitope V in vivo. Deletion of the H-2Db-restricted epitopes I and II/III from T antigen did not increase target cell lysis by epitope V-specific CTL clones in vitro. The amino acid sequence SMIKNLEYM, which species an optimized H-2Db binding motif and was found to induce CTL in H-2b mice, did not further reduce epitope V presentation in vitro when inserted within T antigen. Epitope V-containing T-antigen derivatives which retained epitopes I and II/III or epitope IV did not induce epitope V-specific CTL in vivo: T-antigen derivatives in which epitope V replaced epitope I failed to induce epitope V specific CTL. Recognition of epitope V-H-2Db complexes by multiple independently derived epitope V-specific CTL clones was rapidly and dramatically reduced by incubation of target cells in the presence of brefeldin A compared with the recognition of the other T-antigen CTL epitopes by epitope specific CTL, suggesting that the epitope V-H-2Db complexes either are labile or are present at the cell surface at reduced levels. Our results suggest that processing and presentation of epitope V is not dramatically altered (reduced) by the presence of immunodominant CTL epitopes in T antigen and that the immunorecessive nature of epitope V is not determined by amino acids which flank its native location within simian virus 40 T antigen. PMID- 7474077 TI - Mucosal model of immunization against human immunodeficiency virus type 1 with a chimeric influenza virus. AB - Previously, we constructed a chimeric influenza virus that expresses the highly conserved amino acid sequence ELDKWA of gp41 of human immunodeficiency virus type 1 (HIV-1). Antisera elicited in mice by infection with this chimeric virus showed neutralizing activity against distantly related HIV-1 isolates (T. Muster, R. Guinea, A. Trkola, M. Purtscher, A. Klima, F. Steindl, P. Palese, and H. Katinger, J. Virol. 68:4031-4034, 1994). In the present study, we demonstrated that intranasal immunizations with this chimeric virus are also able to induce a humoral immune response at the mucosal level. The immunized mice had ELDKWA specific immunoglobulins A in respiratory, intestinal, and vaginal secretions. Sustained levels of these secretory immunoglobulins A were detectable for more than 1 year after immunization. The results show that influenza virus can be used to efficiently induce secretory antibodies against antigens from foreign pathogens. Since long-lasting mucosal immunity in the genital and intestinal tracts might be essential for protective immunity against HIV-1, influenza virus appears to be a promising vector for HIV-1-derived immunogens. PMID- 7474078 TI - Genetic analysis of human immunodeficiency virus type 1 integrase and the U3 att site: unusual phenotype of mutants in the zinc finger-like domain. AB - Retroviral integration is the step which leads to establishment of the provirus, cis- and trans-acting regions of the human immunodeficiency type 1 (HIV-1) retrovirus genome, including the attachment site (att) at the ends of the unintegrated viral DNA and the conserved domains within the integrase (IN) protein, have been identified as being important for integration. We investigated the role of each of these regions in the context of an infectious HIV-1 molecular clone through point mutagenesis of the att site and the zinc finger-like and catalytic domains of IN. The effect of each mutation on integration activity was examined by using a single-step infection system with envelope-pseudotype virus. The relative integration efficiency was estimated by monitoring the levels of viral DNA over time in the infected cells. The integration activities of catalytic domain point mutants and att site deletion mutants were estimated to be 0.5 and 5% of wild-type activity, respectively. However, in contrast with previous in vitro cell-free integration studies, alteration of the highly conserved CA dinucleotide resulted in a mutant which still retained 40% of wild type integration activity. The relative levels of expression of each mutant, as measured by a luciferase reporter gene, correlated with levels of integration. This observation is consistent with those of previous studies indicating that integration is an obligatory step for retroviral gene expression. Interestingly, we found that three different HIV-1 constructs bearing point mutations in the zinc finger-like domain synthesized much lower levels of viral DNA after infection, suggesting impairment of these mutants before or at the initiation of reverse transcription. Western blot (immunoblot) analysis demonstrated wild-type levels of reverse transcriptase within the mutant virions. In vitro endogenous reverse transcription assays indicated that all three mutants in the zinc finger like domain had wild-type levels of reverse transcriptase activity. These data indicate that in addition to integration, IN may have an effect on the proper course of events in the viral life cycle that precede integration. PMID- 7474079 TI - Cytomegalovirus infection induces high levels of cyclins, phosphorylated Rb, and p53, leading to cell cycle arrest. AB - Human cytomegalovirus (HCMV) infection stimulates cellular DNA synthesis and causes chromosomal damage. Because such events likely affect cellular proliferation, we investigated the impact of HCMV infection on key components of the cell cycle. Early after infection, HCMV induced elevated levels of cyclin E, cyclin E-associated kinase activity, and two tumor suppressor proteins, p53 and the retinoblastoma gene product (Rb). The steady-state concentration of Rb continued to rise throughout the infection, with most of the protein remaining in the highly phosphorylated form. At early times, HCMV infection also induced cyclin B accumulation, which was associated with a significant increase in mitosis-promoting factor activity as the infection progresses. In contrast, the levels of cyclin A and cyclin A-associated kinase activity increased only at late times in the infection, and the kinetics were delayed relative to those for cyclins E and B. Analysis of the cellular DNA content in the infected cells by flow cytometry showed a progressive shift of the cells from the G1 to the S and G2/M phases of the cell cycle, leading to an accumulation of aneuploid cells at late times. We propose that these HCMV-mediated perturbations result in cell cycle arrest in G2/M. PMID- 7474080 TI - Human immunodeficiency virus type 1 viral protein R (Vpr) arrests cells in the G2 phase of the cell cycle by inhibiting p34cdc2 activity. AB - The Vpr accessory gene product of human immunodeficiency virus types 1 and 2 and simian immunodeficiency virus is believed to play a role in permitting entry of the viral core into the nucleus of nondividing cells. A second role for Vpr was recently suggested by Rogel et al. (M. E. Rogel, L. I. Wu, and M. Emerman, J. Virol. 69:882-888, 1995), who showed that Vpr prevents the establishment in vitro of chronically infected HIV producer cell lines, apparently by causing infected cells to arrest in the G2/M phase of the cell cycle. In cycling cells, progression from G2 to M phase is driven by activation of the p34cdc2/cyclin B complex, an event caused, in part, by dephosphorylation of two regulatory amino acids of p34cdc2 (Thr-14 and Tyr-15). We show here that Vpr arrests the cell cycle in G2 by preventing the activation of the p34cdc2/cyclin B complex. Vpr expression in cells caused p34cdc2 to remain in the phosphorylated, inactive state, p34cdc2/cyclin B complexes immunoprecipitated from cells expressing Vpr were almost completely inactive in a histone H1 kinase assay. Coexpression of a constitutively active mutant p34cdc2 molecule with Vpr relieved the G2 arrest. These findings strongly suggest that Vpr arrests cells in G2 by preventing the activation of the p34cdc2/cyclin B complex that is required for entry into M phase. In vivo, Vpr might, by preventing p34cdc2 activation, delay or prevent apoptosis of infected cells. This would increase the amount of virus each infected cell produced. PMID- 7474081 TI - Quantitative measurement of paramyxovirus fusion: differences in requirements of glycoproteins between simian virus 5 and human parainfluenza virus 3 or Newcastle disease virus. AB - To compare the requirements for paramyxovirus-mediated cell fusion, the fusion (F) and hemagglutinin-neuraminidase (HN) glycoproteins of simian virus 5 (SV5), human parainfluenza virus 3 (HPIV-3), and Newcastle disease virus (NDV) were expressed individually or coexpressed in either homologous or heterologous combinations in CV-1 or HeLa-T4 cells, using the vaccinia virus-T7 polymerase transient expression system. The contribution of individual glycoproteins in virus-induced membrane fusion was examined by using a quantitative assay for lipid mixing based on the relief of self-quenching (dequenching) of fluorescence of the lipid probe octadecyl rhodamine (R18) and a quantitative assay for content mixing based on the cytoplasmic activation of a reporter gene, beta galactosidase. In these assays, expression of the individual F glycoproteins did not induce significant levels of cell fusion and no cell fusion was observed in experiments when cells individually expressing homologous F or HN proteins were mixed. However, coexpression of homologous F and HN glycoproteins resulted in extensive cell fusion. The kinetics of fusion were found to be very similar for all three paramyxoviruses studied. With NDV and HPIV-3, no cell fusion was detected when F proteins were coexpressed with heterologous HN proteins or influenza virus hemagglutinin (HA). In contrast, SV5 F protein exhibited a considerable degree of fusion activity when coexpressed with either NDV or HPIV-3 HN or with influenza virus HA, although the kinetics of fusion were two- to threefold higher when the homologous SV5 F and HN proteins were coexpressed. Thus, these data indicate that among the paramyxoviruses tested, SV5 has different requirements for cell fusion. PMID- 7474082 TI - The role of CD8+ T lymphocytes in coxsackievirus B3-induced myocarditis. AB - Coxsackievirus infections have previously been shown to cause acute or chronic myocarditis in humans, and several mouse models have been established to study the pathology of this disease. Myocardial injury may result from direct viral effects and/or may be immune mediated. To determine the relative roles of these processes in pathogenesis, we have compared coxsackievirus B3 (CVB3) infections of normal and immuno-compromised transgenic knockout (ko) mice. CVB3 was able to infect all strains used (C57BL/6, CD4ko, and beta-microglobulin ko [beta 2Mko]), and following intraperitoneal injection, two disease processes could be distinguished. First, the virus caused early (3 to 7 days postinfection) death in a viral dose-dependent manner. Immunocompetent C57BL/6 mice were highly susceptible (50% lethal dose = 70 PFU), while immunodeficient transgenic ko mice were less susceptible, showing 10- and 180-fold increases in the 50% lethal dose (for CD4ko and beta 2Mko mice, respectively). Second, a histologic examination of surviving CD4ko mice at 7 days postinfection revealed severe myocarditis; the inflammatory infiltrate comprised 40 to 50% macrophages, 30 to 40% NK cells, and 10 to 20% CD8+ T lymphocytes. The infiltration resolved over the following 2 to 3 weeks, with resultant myocardial fibrosis. In vivo depletion of CD8+ T lymphocytes from these CD4ko mice led to a marked reduction in myocarditis and an increase in myocardial virus titers. beta 2Mko mice, which lack antiviral CD8+ T cells, are much less susceptible to early death and to the development of myocarditis. We conclude that our data support a strong immunopathologic component in CVB3-induced disease and implicate both CD4+ and CD8+ T cells. Compared with immunocompetent animals, (i) mice lacking CD4+ T cells (CD4ko) were more resistant to virus challenge, and (ii) mice lacking CD8+ T cells (beta 2Mko and in vivo-depleted CD4ko) showed enhanced survival and a reduced incidence of the later myocarditis. Nevertheless, the picture is complex, since (iii) removal of the CD4+ component, while protecting against early death, greatly magnified the severity of myocarditis, and (iv) removal of the CD8+ cells from CD4ko mice, although protecting against early death and later myocarditis, led to markedly increased virus titers in the heart. These data underscore the complex balance between the costs and benefits of effective antiviral immune responses. PMID- 7474083 TI - Double-stranded RNA bacteriophage phi 6 protein P4 is an unspecific nucleoside triphosphatase activated by calcium ions. AB - Double-stranded RNA bacteriophage phi 6 has an envelope surrounding the nucleocapsid (NC). The NC is composed of a surface protein, P8, and proteins P1, P2, P4, and P7, which form a dodecahedral polymerase complex enclosing the segmented viral genome. Empty polymerase complex particles (procapsids) package positive-sense viral single-stranded RNAs provided that energy is available in the form of nucleoside triphosphates (NTPs). Photoaffinity labelling of both the NC and the procapsid has earlier been used to show that ATP binds to protein P4 and that the NC hydrolyzes NTPs. Using the NC and the NC core particles (NCs lacking surface protein P8) and purified protein P4, we demonstrate here that multimeric P4 is the active NTPase. Isolation of multimeric P4 is successful only in the presence of NTPs. The activity of P4 is the same in association with the viral particles as it is in pure form. P4 is an unspecific NTPase hydrolyzing ribo-NTPs, deoxy NTPs, and dideoxy NTPs to the corresponding nucleoside diphosphates. The Km of the reaction for ATP, GTP, and UTP is around 0.2 to 0.3 mM. The NTP hydrolysis by P4 absolutely requires residual amounts of Mg2+ ions and is greatly activated when the Ca2+ concentration reaches 0.5 mM. Competition experiments indicate that Mg2+ and Ca2+ ions have approximately equal binding affinities for P4. They might compete for a common binding site. The nucleotide specificity and enzymatic properties of the P4 NTPase are similar to the NTP hydrolysis reaction conditions needed to translocate and condense the viral positive-sense RNAs to the procapsid particle. PMID- 7474084 TI - A single retroviral gag precursor signal peptide recognized by FBL-3 tumor specific cytotoxic T lymphocytes. AB - Several dominant T-cell receptors of cytotoxic T-lymphocyte (CTL) clones specific for FBL-3 tumor antigen were clonally amplified in mixed lymphocyte tumor cell cultures derived from an individual immune mouse. Every CTL clone analyzed had a common specificity for a single epitope in the precursor to cell membrane associated nonstructural gag-encoded protein, Pr75gag, which can be minimally identified by nine amino acid residues, SIVLCCLCL. This epitope is located within the hydrophobic signal sequence motif that mediates translocation of the protein into the endoplasmic reticulum. These novel observations suggest that expression of Pr75gag in FBL-3 tumor cells led to the amplification of CTLs which recognize the signal sequence of the nonstructural gag-encoded glycoprotein precursor. PMID- 7474085 TI - Ribosomal frameshifting during translation of measles virus P protein mRNA is capable of directing synthesis of a unique protein. AB - Members of the Paramyxoviridae family utilize a variety of different strategies to increase coding capacity within their P cistrons. Translation initiation at alternative 5'-proximal AUG codons is used by measles virus (MV) to express the virus-specific P and C proteins from overlapping reading frames on their mRNAs. Additional species of mRNAs are transcribed from the MV P cistron by the insertion of extra nontemplated G residues at a specific site within the P transcript. Addition of only a single nontemplated G residue results in the expression of the V protein, which contains a unique carboxyl terminus. We have used an Escherichia coli system to express MV P cistron-related mRNAs and proteins. We have found that ribosomal frameshifting on the MV P protein mRNA is capable of generating a previously unrecognized P cistron-encoded protein that we have designated R. Some ribosomes which have initiated translation of the P protein mRNA use the sequence TCC CCG AG (24 nucleotides upstream of the V protein stop codon) to slip into the -1 reading frame, thus translating the sequence as TC CCC GAG. The resulting R protein terminates five codons downstream of the frameshift site at the V protein stop codon. We have gone on to use a chloramphenicol acetyltransferase reporter system to demonstrate that this MV specific sequence is capable of directing frameshifting during in vivo translation in eukaryotic cells. Analysis of immunoprecipitated proteins from MV infected cells by two-dimensional gel electrophoresis allowed detection of a protein species consistent with R protein in MV-infected cells. Quantitation of this protein species allowed a rough estimation of frameshift frequency of approximately 1.8%. Significant stimulation of ribosomal frameshift frequency at this locus of the MV P mRNA was mediated by a downstream stimulator element which, although not yet fully defined, appeared to be neither a conventional stem loop nor an RNA pseudoknot structure. PMID- 7474086 TI - Characterization of a 100-kilodalton binding protein for the six serotypes of coxsackie B viruses. AB - Viral infection of host cells primarily depends on binding of the virus to a specific cell surface protein. In order to characterize the binding protein for group B coxsackieviruses (CVB), detergent-solubilized membrane proteins of different cell lines were tested in virus overlay protein-binding assays. A prominent virus-binding protein with a molecular mass of 100 kDa was detected in various CVB-permissive human and monkey cell lines but was not detected in nonpermissive cell lines. The specificity of CVB binding to the 100-kDa protein on permissive human cells was substantiated by binding of all six serotypes of CVB and by competition experiments. In contrast, poliovirus and Sendai virus did not bind to the 100-kDa CVB-specific protein. A fraction of HeLa membrane proteins enriched in the range of 100 kDa showed functional activity by transforming infectious CVB (160S) into A-particles (135S). In order to purify this CVB-binding protein, solubilized membrane proteins from HeLa cells were separated by preparative sodium dodecyl sulfate-polyacrylamide gel electrophoresis followed by elution of the 100-kDa protein. Amino acid sequence analysis of tryptic fragments of the CVB-binding protein indicated that this 100 kDa CVB-specific protein is a cell surface protein related to nucleolin. These results were confirmed by immunoprecipitations of the CVB-binding protein with nucleolin-specific antibodies, suggesting that a nucleolin-related membrane protein acts as a specific binding protein for the six serotypes of CVB. PMID- 7474088 TI - Neuronal expression of human immunodeficiency virus type 1 env proteins in transgenic mice: distribution in the central nervous system and pathological alterations. AB - It is now well documented that human immunodeficiency virus type 1 (HIV-1) induces encephalopathy in patients with AIDS. In vitro studies have implicated the envelope protein (gp120) as a factor which causes neuronal death. To better evaluate the role and elucidate the mechanisms of gp120 neurotoxicity, we have developed transgenic mice carrying a segment of the HIV-1 genome that expresses the viral gp160 protein under the control of the human neurofilament light gene promoter. In two separate lines of transgenic mice, the Env protein was found to be expressed in several nuclei of the brain stem and in the anterior horns of the spinal cord. The two lines showed identical patterns of Env expression. Neuropathological evaluation revealed numerous abnormal dendritic swellings in the immunostained motor neuron structures. Large and numerous neuritic swellings were also prominent in the nucleus gracilis and in the gracilis and cuneate fascicles. In addition, reactive astrocytosis was observed in several immunoreactive areas of the central nervous system. These transgenic mice offer a unique model to further investigate the role of HIV-1 Env protein in neuronal toxicity and to help elucidate the mechanisms that are involved. PMID- 7474087 TI - Functional characterization of human immunodeficiency virus type 1 nef genes in patients with divergent rates of disease progression. AB - We have studied the sequence and function of the human immunodeficiency virus type 1 (HIV-1) nef genes from nine patients with highly divergent rates of disease progression enrolled in a longitudinal study of HIV disease. Over an average of 7.8 years of follow-up, three patients had net positive changes in CD4+ T-cell counts, three patients had net negative changes in CD4+ T cells but did not develop AIDS, and three patients progressed to AIDS. The nef gene from each of these patients was amplified and cloned, and the sequence of 8 to 10 clones was determined. Only 2 of 88 (2.3%) nef genes recovered from these nine patients were grossly defective. Moreover, there was no relationship between the phylogeny of nef sequences and the corresponding rates of disease progression from these patients. Representative nef genes from all nine patients were tested for their abilities to downregulate cell surface CD4 in a transient-transfection assay. There was no correlation found between the functions of the nef genes from these patients and their corresponding rates of disease progression. We conclude that the nef gene is not a common mediator of the rate of HIV disease progression in natural infection. PMID- 7474089 TI - Role of the virion host shutoff (vhs) of herpes simplex virus type 1 in latency and pathogenesis. AB - The herpes simplex virus type 1 (HSV-1) UL41 gene product, virion host shutoff (vhs), has homologs among five alphaherpesviruses (HSV-1, HSV-2, pseudorabies virus, varicella-zoster virus, and equine herpesvirus 1), suggesting a role for this protein in neurotropism. A mutant virus, termed UL41NHB, which carries a nonsense linker in the UL41 open reading frame at amino acid position 238 was generated. UL41NHB and a marker-rescued virus, UL41NHB-R, were characterized in vitro and tested for their ability to replicate in vitro and in vivo and to establish and reactivate from latency in a mouse eye model. As demonstrated by Western blotting (immunoblotting) and Northern (RNA) blotting procedures, UL41NHB encodes an appropriately truncated vhs protein and, as expected for a vhs null mutant, fails to induce the degradation of cellular glyceraldehyde-3-phosphate dehydrogenase mRNA. The growth of UL41NHB was not significantly altered in one step growth curves in Vero or mouse C3H/10T1/2 cells but was impaired in corneas, in trigeminal ganglia, and in brains of mice compared with the growth of KOS and UL41NHB-R. As a measure of establishment of latency, quantitative DNA PCR showed that the amount of viral DNA within trigeminal ganglia latently infected with UL41NHB was reduced by approximately 30-fold compared with that in KOS-infected ganglia and by 50-fold compared with that in UL41NHB-R-infected ganglia. Explant cocultivation studies revealed a low reactivation frequency for UL41NHB (1 of 28 ganglia, or 4%) compared with that for KOS (56 of 76, or 74%) or UL41NHB-R (13 of 20 or 65%). Taken together, these results demonstrate that vhs represents a determinant of viral pathogenesis. PMID- 7474091 TI - Fast and high-affinity binding of B-lymphotropic papovavirus to human B-lymphoma cell lines. AB - Binding of B-lymphotropic papovavirus (LPV) to host cells differing in susceptibility to viral infection was determined by a newly established, direct, nonradioactive virus binding assay, which allows quantitative description of the binding characteristics by receptor saturation and Scatchard analysis. LPV binding to the highly susceptible human B-lymphoma cell line BJA-B K88 is specific, saturable, and noncooperative. Binding occurs very fast, with an association rate constant (k1) of 6.7 x 10(7) M-1s-1, and is of high affinity, with a dissociation constant (Kd) of 2.9 x 10(-12) M; and the virus-receptor complex is stable, with a half life of 70 min. The binding affinities of receptors on four other highly, moderately, or weakly susceptible human B lymphoma cell lines were similar, with up to twofold variation around a mean Kd value of 3 x 10(-12) M, suggesting the presence of the same LPV receptor on all of these cell lines. This view is further supported by the finding that in all cases a terminal sialic acid is necessary for LPV binding. Tunicamycin has been shown to drastically induce LPV susceptibility and LPV binding in weakly and moderately susceptible B-lymphoma cell lines (O.T. Keppler, M. Herrmann, M. Oppenlander, W. Meschede, and M. Pawlita, J. Virol. 68:6933-6939, 1994). The hypothesis that the constitutively expressed and tunicamycin-induced LPV receptors are identical is strengthened by our finding that both receptor types displayed the same high affinity. LPV susceptibility of different B-lymphoma cell lines was correlated with receptor number but not with receptor affinity. The numbers of receptors per cell on highly and moderately susceptible cell lines ranged from 2,000 to 400 and were directly proportional to LPV susceptibility. This indicates that the number of high-affinity receptors per cell is a key regulating factor for the LPV host range. PMID- 7474090 TI - Negative regulation of the adeno-associated virus (AAV) P5 promoter involves both the P5 rep binding site and the consensus ATP-binding motif of the AAV Rep68 protein. AB - Transcript levels from the P5 promoter of adeno-associated virus type 2 (AAV) are negatively regulated by the AAV Rep78 and Rep68 proteins in the absence of helper virus. We have identified a Rep-responsive negative cis element of the P5 promoter between the P5 TATA box and transcription start site by using 5' and 3' deletions of the P5 promoter fused to the chloramphenicol acetyltransferase gene. This element contains four imperfect GAGC repeats similar to the Rep recognition sequences (RRSs) in the AAV inverted terminal repeats and in the AAV preferred integration locus in chromosome 19. Band shift analyses showed that human 293 cell nuclear extracts containing Rep68 or Rep68/K340H, a putative nucleoside triphosphate (NTP)-binding-site mutant of Rep68, formed Rep-specific complexes with this P5 RRS DNA. Within the P5 RRS, mutation of a cytosine at position 273 in the AAV sequence to guanine abolished Rep68 binding to the DNA. A mutation in the P5 RRS within a full-length AAV genome, which abolished Rep binding, resulted in a 40 to 50% reduction in the ability of wild-type Rep68 to inhibit the accumulation of P5 transcripts in vivo. In contrast, the Rep68/K340H mutant was unable to down-regulate this mutated promoter. These results indicate that there are at least two mechanisms involved in the negative regulation of P5 transcript levels by Rep68; one involves Rep68 binding to the P5 RRS, and another requires the region of Rep68 containing the consensus NTP-binding motif. Furthermore, our studies of AAV genomes containing mutated RRS- and/or YY1-binding elements suggest that transcription factor YY1 binding to the transcription start site of P5 interferes with Rep68 repression of the P5 promoter. PMID- 7474092 TI - Extreme fitness differences in mammalian and insect hosts after continuous replication of vesicular stomatitis virus in sandfly cells. AB - Continuous, persistent replication of a wild-type strain of vesicular stomatitis virus in cultured sandfly cells for 10 months profoundly decreased virus replicative fitness in mammalian cells and greatly increased fitness in sandfly cells. After persistent infection of sandfly cells, fitness was over 2,000,000 fold greater than that in mammalian cells, indicating extreme selective differences in the environmental conditions provided by insect and mammalian cells. The sandfly-adapted virus also showed extremely low fitness in mouse brain cells (comparable to that in mammalian cell cultures). It also showed an attenuated phenotype, requiring a nearly millionfold higher intracranial dose than that of its parent clone to kill mice. A single passage of this adapted virus in BHK-21 cells at 37 degrees C restored fitness to near neutrality and also restored mouse neurovirulence. These results clearly illustrate the enormous capacity of RNA viruses to adapt to changing selective environments. PMID- 7474093 TI - p6Gag is required for particle production from full-length human immunodeficiency virus type 1 molecular clones expressing protease. AB - The human immunodeficiency virus type 1 (HIV-1) Gag protein precursor, Pr55Gag, contains at its C-terminal end a proline-rich, 6-kDa domain designated p6. Two functions have been proposed for p6: incorporation of the HIV-1 accessory protein Vpr into virus particles and virus particle production. To characterize the role of p6 in the HIV-1 life cycle and to map functional domains within p6, we introduced a number of nonsense and single and multiple amino acid substitution mutations into p6. Following the introduction of the mutations into the full length HIV-1 molecular clone pNL4-3, the effects on Gag protein expression and processing, virus particle production, and virus infectivity were analyzed. The production of mutant virus particles was also examined by transmission electron microscopy. The results indicate that (i) p6 is required for efficient virus particle production from a full-length HIV-1 molecular clone; (ii) a Pro-Thr-Ala Pro sequence, located between residues 7 and 10 of p6, is critical for virus particle production; (iii) mutations outside the Pro-Thr-Ala-Pro motif have little or no effect on virus assembly and release; (iv) the p6 defect is manifested at a late stage in the budding process; and (v) mutations in p6 that severely reduce virion production in HeLa cells also block or significantly delay the establishment of a productive infection in the CEM (12D-7) T-cell line. We further demonstrate that mutational inactivation of the viral protease reverses the p6 defect, suggesting a functional linkage between p6 and the proteolytic processing of the Gag precursor protein during the budding of progeny virions. PMID- 7474094 TI - Cell proteins bind to a linear polypyrimidine-rich sequence within the 5' untranslated region of rhinovirus 14 RNA. AB - Members of the picornavirus family initiate translation of their RNA genomes by a cap-independent mechanism in which ribosomes bind to an internal site in the 5' untranslated region (5'-UTR). This unique process requires an internal ribosome entry site (IRES), a highly structured RNA whose function is mediated in part by interactions with cell proteins. The IRES element of human rhinovirus 2 (HRV-2) extends from nucleotide (nt) 10 to between nt 544 and 568 and has been shown to interact with two cell proteins, pyrimidine tract-binding protein (pPTB) and p97. To map the specific regions of HRV-14 RNA that bind cell proteins, mobility shift, UV cross-linking and Western immunoblot analyses were performed. The results indicate that an RNA sequence from nt 538 to 591 interacts with pPTB and La, two proteins previously shown to functionally interact with the IRES elements of several picornaviruses. Two additional proteins, p97 and p68, were also cross linked to nt 538 to 591 RNA. These four proteins interact with a putatively unstructured portion of the 5'-UTR that contains a polypyrimidine tract and has been shown to be present at the 3' border of sequences that are essential for IRES function of HRV-2. These protein-RNA interactions are likely to play a role in internal initiation of translation. PMID- 7474095 TI - A new cis-acting element for RNA replication within the 5' noncoding region of poliovirus type 1 RNA. AB - Mouse cells expressing the human poliovirus receptor (PVR-mouse cells) as well as human HeLa cells are susceptible to poliovirus type 1 Mahoney strains and produce a large amount of progeny virus at 37 degrees C. However, the virus yield is markedly reduced at 40 degrees C in PVR-mouse cells but not in HeLa cells. The reduction in virus yield at 40 degrees C appears to be due to a defective initiation process in positive-strand RNA synthesis (K. Shiroki, H. Kato, S. Koike, T. Odaka, and A. Nomoto, J. Virol. 67:3989-3996, 1993). To gain insight into the molecular mechanisms involved in this detective process, naturally occurring heat-resistant (Hr)-mutants which show normal growth ability in PVR mouse cells even at 40 degrees C were isolated from a virus stock of the Mahoney strain and their mutation sites that affect the phenotype were identified. The key mutation was a change from adenine (A) to guanine (G) at nucleotide position (nt) 133 within the 5' noncoding region of the RNA. This mutation also gave an Hr phenotype to the viral plus-strand RNA synthesis in PVR-mouse cells. Mutant Mahoney strains with a single point mutation at nt 133 (A to G, C, or T or deletion) were investigated for their ability to grow in PVR-mouse cells at 40 degrees C. Only the mutant carrying G at nt 133 showed an Hr growth phenotype in PVR-mouse cells. These results suggest that a host cellular factor(s) interacts with an RNA segment around nt 133 of the plus-strand RNA or the corresponding region of the minus-strand RNA, contributing to efficiency of plus-strand RNA synthesis. PMID- 7474096 TI - Insertions within the hepatitis B virus capsid protein influence capsid formation and RNA encapsidation. AB - Hepatitis B virus (HBV) capsid proteins, termed core proteins, with two- to four amino-acid insertions were assessed for capsid formation, RNA encapsidation, and the ability to support reverse transcription of the pregenome by the polymerase molecule. Velocity sedimentation analysis of insect cell-expressed recombinant core proteins revealed that only two of the nine insertion mutant proteins formed capsids with the tight banding patterns of wild-type capsids. The remaining mutant core proteins were spread over the gradients, suggesting aggregate formation, or at the top of the gradients, suggesting lack of stable capsid formation. The mutant capsid proteins were coexpressed in Huh7 cells with an HBV genome lacking a functional core gene to test for trans complementation of HBV replication. Three of the mutant core proteins formed capsids containing HBV RNA, but only two of these contained reverse-transcribed HBV DNA. While the core protein has shown resiliency in capsid formation following insertion of foreign residues into the major B-cell epitope, several of the small insertions severely reduced the efficiency of capsid formation and inhibited capsid function. PMID- 7474097 TI - E- vectors: development of novel self-inactivating and self-activating retroviral vectors for safer gene therapy. AB - We have developed novel self-inactivating and self-activating retroviral vectors based on the previously observed high-frequency deletion of direct repeats. We constructed spleen necrosis virus (SNV)-based viral vectors that contained large direct repeats flanking the viral encapsidation sequence (E). A large proportion of the proviruses in the target cells had E and one copy of the direct repeat deleted. Direct repeats of 1,333 and 788 bp were deleted at frequencies of 93 and 85%, respectively. To achieve a 100% deletion efficiency in target cells after ex vivo infection and drug selection, we constructed a self-activating vector that simultaneously deleted E and reconstituted the neomycin phosphotransferase gene. Selection of the target cells for resistance to G418 (a neomycin analog) ensured that all integrated proviruses had E deleted. The proviruses with E deleted were mobilized by a replication-competent virus 267,000-fold less efficiently than proviruses with E. We named these self-inactivating vectors E- (E-minus) vectors. These vectors should increase the safety of retroviral vector-mediated gene therapy by preventing the spread of vector sequences to nontarget cells in the event of coinfection with helper virus. We propose that direct-repeat deletions occur during RNA-dependent DNA synthesis and suggest that template switches occur without a requirement for RNA breaks. The minimum template dissociation frequency was estimated as 8%/100 bp per replication cycle. These vectors demonstrate that large direct repeats and template-switching properties of reverse transcriptase can be utilized to delete any sequence or reconstitute genes during retroviral replication. PMID- 7474098 TI - Immunogenic determinants of a neuropathogenic murine leukemia virus. AB - Previous studies of Cas-Br-M murine leukemia virus (MuLV) (Cas-MuLV) infection demonstrated that cytotoxic T cells (CTL) of the CD8+ phenotype play a role in resistance to the neuropathogenic effects of the virus in NFS/N mice. In the current study, we sought to identify the Cas-MuLV epitopes that are immunogenic for the CTL response. Infection of adult NFS/N mice with a well-characterized neuropathogenic variant of Friend MuLV, PVC-211 MuLV (PVC-MuLV), was not immunogenic for MuLV-specific CTL. Therefore, we constructed chimeric viruses between Cas-MuLV and PVC-MuLV. Infectious chimeras contained the Cas-MuLV env gene on a PVC-MuLV background (PVC-CasenvMuLV) and the PVC-MuLV env gene on a Cas MuLV background (Cas-PVCenvMuLV). Cas-MuLV-specific CTL were found following inoculation of both the chimeric viruses and the parental Cas-MuLV but not the parental PVC-MuLV, despite evidence of antibody responses to both parental and chimeric MuLV. CTL generated in response to infection with PVC-CasenvMuLV and Cas PVCenvMuLV were exclusively of the CD8+ phenotype. These results indicate that both the env and gag-pol regions of Cas-MuLV express epitopes that are immunogenic for CTL. PMID- 7474100 TI - Human immunodeficiency virus type 1 Vpr arrests the cell cycle in G2 by inhibiting the activation of p34cdc2-cyclin B. AB - Human immunodeficiency virus type 1 (HIV-1) vpr inhibits the replication of tumor cell lines and peripheral blood mononuclear cells. Here it is demonstrated that expression of vpr, either in the context of a provirus or from an independent genetic element, induces a discrete cell cycle arrest, with cells containing 4N DNA. Low cyclin B-associated kinase activity, as well as the status of p34cdc2 and cdc25C phosphorylation, indicates that the cascade of reactions which drives the cell into mitosis has not been initiated. The phosphatase inhibitor okadaic acid releases the block, suggesting that Vpr perturbs upstream regulatorsof the G2-M transition. These studies demonstrate that HIV-1 vpr has profound effects on the cellular factors which control entry into mitosis and indicate vpr's potential contribution to the cellular pathology associated with HIV-1 infection. PMID- 7474099 TI - Polypyrimidine tract-binding protein and heterogeneous nuclear ribonucleoprotein A1 bind to human T-cell leukemia virus type 2 RNA regulatory elements. AB - Efficient expression of human T-cell leukemia virus (HTLV) and human immunodeficiency virus structural proteins requires Rx and Rev proteins, respectively. Decreased expression of Gag and Env appears to be due, in part, to intragenic RNA sequences, termed cis-acting repressive sequences (CRS), and may be mediated by binding of specific cellular factors. We demonstrated previously that two cellular proteins, p60CRS and p40CRS, interact with HTLV type 2.5' long terminal repeat CRS RNA and that the interaction of both proteins with CRS RNA correlates with function (A. C. Black, C. T. Ruland, J. Luo, A. Bakker, J. K. Fraser, and J. D. Rosenblatt, Virology 200:29-41, 1994). By radioimmunoprecipitation of HeLa nuclear proteins UV cross-linked to CRS RNAs with murine monoclonal antibodies, we now show that p40CRS is heterogeneous nuclear ribonucleoprotein (hnRNP) A1 and p60CRS is polypyrimidine tract-binding protein or hnRNP I. These immunoprecipitation results were confirmed by an immunobinding assay with hnRNP I and hnRNP AI antibodies and by cross-competition electrophoretic mobility shift experiments. In addition, we mapped a putative hnRNP A1 binding site in U5 RNA and demonstrated that p40CRS (hnRNP A1) binding to that site correlates with CRS function. Since both hnRNP I and hnRNP A1 have been shown to influence splicing and potentially other steps in RNA processing, the binding of both hnRNP I and hnRNP A1 to HTLV RNA regulatory elements may alter retrovirus RNA processing and may be involved in regulation by Rex. PMID- 7474101 TI - Differential regulation of human papillomavirus type 6 and 11 early promoters in cultured cells derived from laryngeal papillomas. AB - Cells cultured from laryngeal papillomas contain episomal human papillomavirus type 6 or type 11 (HPV-6/11) DNA. We developed a sensitive RNase protection assay to simultaneously measure expression from the HPV E6, E7, and E1 promoters (P1, P2 and P3, respectively) in this manipulable culture system and found that P1, P2 and P3 transcript abundances could be independently modulated by culture medium composition and culture substrate. In undifferentiated cells grown in a low calcium, serum-free medium, P1 transcripts commonly predominated over those from P2, P3 transcripts were often undetectable, and high concentrations of retinoic acid were able to selectively decrease P2 transcript abundance. When cultures were allowed to stratify and differentiate by growth on a collagen gel at he air liquid interface, total HPV RNA increased up to sixfold because of selective increases in abundances of P1 and P3 transcripts. High-calcium submerged cultures also showed easily detectable P3 transcripts, and isolated suprabasal cells contained almost exclusively these transcripts. Growth arrest alone was not sufficient to induce P3 transcripts. Thus, in contrast to the HPV-6/11 E6 and E7 promoters, the E1 promoter was utilized primarily in a differentiation-specific manner. We also show that increased HPV gene dosage will not necessary bring about increased HPV transcript abundance, suggesting that other viral and cellular factors are responsible for regulation of total transcript levels as well as specific promoter usage. PMID- 7474102 TI - A leucine triplet repeat sequence (LXX)4 in p6gag is important for Vpr incorporation into human immunodeficiency virus type 1 particles. AB - Incorporation of Vpr into human immunodeficiency virus type 1 (HIV-1) virions is mediated by the Gag protein, independently of other viral components. We have coexpressed Vpr and Gag constructs in a vaccinia virus expression system in order to map the region of Gag involved in Vpr packaging. Deletion of the carboxyl terminal p6 region of Gag impaired the ability of Gag to package Vpr. To confirm the role of p6 in Vpr packaging, Rous sarcoma virus (RSV)-HIV chimeras containing HIV-1 p6 were constructed. Although RSV Gag does not package Vpr into virus particles, a chimera containing HIV-1 p6 is sufficient for Vpr incorporation. To map the region of p6 involved in Vpr packaging, a series of p6 point mutations and deletion mutations was analyzed. Mutations in the N-terminal p6 proline-rich domain, for which preliminary evidence shows a marked decrease in virion incorporated RNA, did not affect Vpr incorporation. Deletion of residues 1 to 31 of HIV-1 p6 did not affect Vpr packaging, but residues 35 to 47, including an (LXX)4 domain, were required for Vpr incorporation into virus particles. PMID- 7474103 TI - High-level expression of adeno-associated virus (AAV) Rep78 or Rep68 protein is sufficient for infectious-particle formation by a rep-negative AAV mutant. AB - Adeno-associated virus (AAV) codes for four closely related nonstructural proteins (Rep) required for AAV DNA replication and gene regulation. In vitro studies have revealed that either Rep78 or Rep68 alone is sufficient for AAV DNA replication. Rep52 and Rep40 are not required for DNA replication but have been reported to enhance the efficiency of accumulation of single-stranded progeny DNA. Previous studies on rep-expressing cell lines had indicated that only a subset of the four Rep proteins are required for the production of infectious AAV. We therefore set out to determine the minimal set of Rep proteins sufficient for the generation of infectious AAV. Transient cotransfections in HeLa cells of constructs for high-level expression of individual Rep proteins with a rep negative AAV genome revealed that either Rep78 or Rep68 alone could complement for a full replication cycle yielding infectious virus. This result was confirmed by transfection studies in the cell line HeM2, which selectively expresses Rep78 at rather low levels under the control of the glucocorticoid-responsive mouse mammary tumor virus long terminal repeat (C. Holscher, M. Horer, J. A. Kleinschmidt, H. Zentgraf, A. Burkle, and R. Heilbronn, J. Virol. 68:7169-7177, 1994). Increasing the level of Rep78 expression by transfection of a glucocorticoid receptor expression construct resulted in a higher level of DNA replication of a cotransfected rep-negative AAV genome and in the production of infectious rep-negative AAV particles. We further report on the generation of a new rep-expressing cell line, HeCM1, which was obtained by stable supertransfection of a construct for constitutive Rep40 expression into HeM1 cells (Holscher et al., J. Virol. 68:7169-7177). Transfection of rather large amounts of rep-negative AAV DNA led to detectable virus production in HeCM1 cells even in the absence of the cotransfected glucocorticoid receptor expression construct, but higher yields were obtained after increasing the Rep78 level by coexpression of the glucocorticoid receptor. These data demonstrate that all Rep functions required for the productive replication of AAV in HeLa cells are contained within both Rep78 and Rep68. PMID- 7474104 TI - Transfer of the minus strand of DNA during hepadnavirus replication is not invariable but prefers a specific location. AB - The current model for replication of duck hepatitis B virus has reverse transcription initiating and copying a UUAC motif within the encapsidation signal, epsilon, near the 5' end of the RNA template. This results in synthesis of four nucleotides of DNA. This short minus-strand DNA product is then transferred to a complementary position, at DR1, near the 3' end of the RNA template. Elongation of minus-strand DNA then ensues. We have examined the transfer of minus-strand DNA during replication of duck hepatitis B virus in cell culture. The initial aim of this work was to examine the effect of mutations at DR1 on the transfer process. We found that when mutations were introduced into the UUAC motif overlapping DR1, the 5' end of minus-DNA no longer mapped to position 2537 but was shifted two or four nucleotides. Mismatches were predicted to exist at the new sites of elongation. Elongation from nucleotide 2537 could be restored in these mutants by making compensatory changes in the UUAC motif within epsilon. This finding led us to examine limitations in the shifting of the site of transfer. When the UUAC motif in epsilon was changed to six different tetranucleotide motifs surrounding position 2537, transfer of minus-strand DNA shifted predictably, albeit inefficiently. Also, when multiple UUAC motifs were introduced near DR1, the UUAC motif at nucleotide 2537 was used preferentially. Overall, our findings confirm the current minus-strand DNA transfer model and demonstrate a marked preference for the site of the transfer. PMID- 7474105 TI - Characterization of double-stranded RNA satellites associated with the Trichomonas vaginalis virus. AB - Three small and distinct satellite double-stranded RNAs (dsRNAs) denoted s1, s1', and s2 were recently described for a Trichomonas vaginalis isolate harboring a dsRNA virus. Since characterization of these satellite dsRNAs might provide insight into the virus replication cycle and virus-host interactions, full-length cDNAs to s1 and s1' dsRNAs were synthesized and sequenced. s1 dsRNA has 688 bp, and s1' dsRNA has 616 bp. A 228-bp open reading frame that begins at nucleotide 37 was detected on a putative sense strand of s1. All satellite RNAs were found associated with RNA-dependent RNA polymerase (RDRP) activity that banded on CsCl gradients. Within carrier trichomonads, satellite RNAs synthesized single stranded replicative intermediates. An in vitro assay was established to assess replication of satellite RNAs. Transcripts generated from s1 cDNA, for example, served as a template for the viral RDRP. These templates had a polarity similar to that of the replicative intermediate found in the satellite-harboring parasites. Importantly, the recognition of s1 RNA was shown to be specific, since unrelated RNAs did not serve as templates for RDRP under the same experimental conditions. The data indicate that the cDNA of s1 has a specific and essential sequence needed for recognition by the viral RDRP and for subsequent RNA synthesis. Both s1 and s1' have conserved domains, albeit of unproven function, but which may be required for replication. PMID- 7474106 TI - Functional epitope analysis of the human CD4 molecule: antibodies that inhibit human immunodeficiency virus type 1 gene expression bind to the immunoglobulin CDR3-like region of CD4. AB - We recently demonstrated that monoclonal antibody (MAb) 13B8-2, specific for the immunoglobulin (Ig) complementary determining region 3 (CDR3)-like region of the CD4 molecule, inhibits viral transcription in human immunodeficiency virus (HIV) infected CEM cells and HIV type 1 (HIV-1) promoter activity. Here, we have studied the capacity of several MAb specific for the D1 domain of CD4, including anti-CDR2-like (Leu-3a and ST4) and anti-CDR3-like (13B8-2 and ST40) MAb, and for the D2 domain of CD4 (BL4) to inhibit both provirus transcription in HIV-1LAI infected CEM cells and transcription of the chloramphenicol acetyltransferase (CAT) gene under control of the HIV-1 long terminal repeat in transiently transfected CEM cells. We found that HIV-1 promoter activity and provirus transcription are inhibited only by MAb that bind to the CDR3-like region in domain 1 of CD4. Moreover, we demonstrated that the Fab fragment of an anti-CDR3 like region-specific anti-CD4 MAb is a powerful inhibitor of HIV-1 promoter activity. These results have implications for understanding the role of the CDR3 like region in CD4 T-cell signaling, which controls provirus transcription. PMID- 7474107 TI - The cytoplasmic tail of CD4 is required for inhibition of human immunodeficiency virus type 1 replication by antibodies that bind to the immunoglobulin CDR3-like region in domain 1 of CD4. AB - Monoclonal antibodies (MAb) directed against the immunoglobulin complementary determining region 3 (CDR3)-like region of the CD4 molecule inhibit human immunodeficiency virus type 1 (HIV-1) transcription. We report here data showing that the cytoplasmic tail of CD4 is required for such inhibition to be achieved. To this aim, we studied the effect of MAb 13B8-2 treatment on (i) HIV-1 production in A2.01 cells, which express different forms of the CD4 gene, (ii) Tat-induced HIV-1 promoter activation, and (iii) mitogen-activated protein kinase (MAPK) activation, which is induced in CD4-positive cells by HIV-1 cross-linking of CD4. Inhibition of HIV production by 13B8-2 MAb treatment was consistently observed in cells expressing wild-type CD4 and cells expressing a hybrid CD4-CD8 molecule (amino acids 1 to 177 of CD4 fused to the hinge, transmembrane, and cytoplasmic domains of CD8). However, no delay in HIV-1 production was observed in cells expressing a truncated CD4 which lacks the cytoplasmic domain (CD4.401). Chloramphenicol acetyltransferase assays demonstrated that Tat-dependent activation of the HIV-1 long terminal repeat promoter was inhibited by MAb 13B8-2 in A2.01/CD4 and A2.01/CD4-CD8 but not in A2.01/CD4.401 cells. Finally, we found that MAb 13B8-2 treatment inhibited the activation of MAPK induced in A2.01/CD4 and A2.01/CD4-CD8 following cross-linking of CD4 by HIV-1. PMID- 7474108 TI - Intrahost human immunodeficiency virus type 1 evolution is related to length of the immunocompetent period. AB - The antigenic diversity threshold theory predicts that antigenic sites of human immunodeficiency virus type 1, such as the V3 region of the external glycoprotein gp120, evolve more rapidly during the symptom-free period in individuals progressing to AIDS than in those who remain asymptomatic for a long time. To test this hypothesis, genomic RNA sequences were obtained from the sera of 44 individuals at seroconversion and 5 years later. The mean number of nonsynonymous nucleotide substitutions in the V3 region of the viruses circulating in 31 nonprogressors (1.1 x 10(-2) +/- 0.1 x 10(-2) per site per year) was higher than the corresponding value for 13 progressors (0.66 x 10(-2) +/- 0.1 x 10(-2) per site per year) (P < 0.01), while no difference between the mean numbers of synonymous substitutions in the two groups was seen (0.37 x 10(-2) +/- 0.1 x 10( 2) and 0.51 x 10(-2) +/- 0.2 x 10(-2) per site per year for nonprogressors and progressors, respectively; P > 0.1). The mean ratios of synonymous nucleotide p distance to nonsynonymous p distance were 0.35 for nonprogressors and 0.62 for progressors. The number of nonsynonymous substitutions was not associated with virus load or virus phenotype, which are established predictors of disease progression, but correlated strongly with the duration of the immunocompetent period (r2 = 0.41; P = 0.001). This indicates that there is no causative relationship between intrahost evolution and CD4+ cell decline. Our data suggest that intrahost evolution in human immunodeficiency virus type 1 infection is driven by selective forces, the strength of which is related to the duration of the immunocompetent period. PMID- 7474109 TI - Recombinant junctions formed by site-specific integration of adeno-associated virus into an episome. AB - A model system using an episomal Epstein-Barr virus shuttle vector was recently developed to study the adeno-associated virus (AAV) site-specific integration event in chromosome 19q13.3-qter (C. Giraud, E. Winocour, and K.I. Berns, Proc. Natl. Acad. Sci. USA 91:10039-10043, 1994). In this study, we analyze the recombinant junctions generated after integration of the AAV genome into an Epstein-Barr virus shuttle vector carrying 8.2, 1.6, or 0.51 kb of the chromosome 19 preintegration sequence (AAVS1 locus). In most of the recombinants, one end of the viral genome was joined to a portion of the AAVS1 DNA previously shown to be a minimum target for AAV integration. Within this AAVS1 segment, the AAV insertion points were strikingly clustered around a binding site for the AAV regulatory protein. In all cases, the second junction with AAV occurred with vector DNA outside of the AAVS1 segment. With respect to the viral genome, one junction with the shuttle vector DNA occurred either within the AAV inverted terminal repeat (itr), or near the P5 promoter, approximately 100 nucleotides distal to a modified itr. The modified itr in 5 of 11 recombinants involved a head-to-tail organization. In one such instance, the AAV insert contained slightly more than one genome equivalent arranged in a head-to-tail manner with a junction close to the P5 promoter; the AAV insert in this recombinant episome could be rescued by adenovirus infection and replicated to virus particles. The significance of the head-to-tail organization is discussed in terms of the possible circularization of AAV DNA before or during integration. PMID- 7474111 TI - A 48-amino-acid region of influenza A virus PB1 protein is sufficient for complex formation with PA. AB - The concerted activity of four influenza virus proteins, PB1, PB2, PA, and NP is necessary and sufficient for transcription and replication of the viral genome in the nucleus of the cell. The three P proteins form a heterotrimeric complex in virions and the nuclei of infected cells. Biochemical analyses have shown specific interactions between PB1 and PA as well as PB1 and PB2, indicating that PB1 is the backbone of the complex. To identify domains of PB1 involved in binding PA, a two-hybrid system adapted for mammalian cells (CV-1) was implemented. First, we demonstrate the ability of PB1 and PA to interact efficiently and specifically in reciprocal combinations of two-hybrid reporter moieties, suggesting that transcription factor module fusion did not interfere sterically or allosterically with interaction between PB1 and PA. Subsequent analyses with a set of chimeric proteins with truncations of the PB1 C termini, N termini, or internal sequences led to the identification of a region at the N terminus of PB1 responsible for binding PA. Forty-eight amino acids at the N terminus of PB1 were sufficient for binding PA in vivo with the same efficiency as the complete PB1 protein. This region of PB1 responsible for binding PA does not overlap with other previously described PB1 functional domains involved in nuclear transport and RNA polymerization. We propose to name this region of interaction with PA domain alpha, to differentiate it from other functional domains described for PB1. PMID- 7474110 TI - Contact of human immunodeficiency virus type 1-infected and uninfected CD4+ T lymphocytes is highly cytolytic for both cells. AB - Individuals infected with the human immunodeficiency virus (HIV) experience a marked loss of CD4+ T lymphocytes, leading to fatal immunodeficiency. The mechanisms causing the depletion of these cells are not yet understood. In this study, we observed that CD4+ T lymphocytes from HIV type 1 (HIV-1)-infected and uninfected individuals rapidly lysed B lymphoblasts expressing the HIV-1 envelope glycoprotein on the cell surface and Jurkat cells expressing the complete virus. Contact of uninfected CD4+ T cells with envelope glycoprotein-expressing cells also resulted in the lysis of the uninfected CD4+ T cells. Cytolysis did not require priming or in vitro stimulation of the CD4+ T cells and was not restricted by major histocompatibility complex molecules. Cytotoxicity was inhibited by soluble CD4 and anti-CD4 monoclonal antibodies that block binding of CD4 to gp120. In addition, neutralizing anti-CD4 and anti-gp120 monoclonal antibodies which block postbinding membrane fusion events and syncytium formation also inhibited cell lysis, suggesting that identical mechanisms in HIV-infected cultures underlie cell-cell fusion and the cytolysis observed. However, cytotoxicity was not always accompanied by the formation of visible syncytia. Rapid cell lysis after contact of uninfected and HIV-1-infected CD4+ T cells may explain CD4+ T-cell depletion in the absence of detectable syncytia in infected individuals. Moreover, because of its vigor, lysis of envelope-expressing targets by contact with unprimed CD4+ T lymphocytes may at first glance resemble antigen specific immune responses and should be excluded when cytotoxic T-lymphocyte responses in infected individuals and vaccinees are evaluated. PMID- 7474112 TI - Evidence that a cell cycle regulator, E2F1, down-regulates transcriptional activity of the human immunodeficiency virus type 1 promoter. AB - Proliferation of eukaryotic cells is orchestrated by a series of cellular proteins which participate in various stages of the cell cycle to guide the cell through mitosis. Some of these proteins, including E2F1, play a critical role in G1 and S phases by coordinately regulating expression of several important cell cycle-associated genes. On the basis of recent observations indicating a block in human immunodeficiency virus type 1 (HIV-1) replication in cells arrested in G1/S phase of the cell cycle, we sought to evaluate the regulatory action of E2F1 on transcription from the HIV-1 long terminal repeat (LTR). Results from transient transfection of cells with an E2F1 expression plasmid indicated that E2F1 has the ability to suppress basal transcriptional activity of the LTR and to diminish the extent of the Tat-induced activation of the viral promoter. Deletion analysis of the HIV-1 LTR in transfection studies revealed the presence of two major elements responsive to E2F1 repression located distally (-454 to -381) and proximally ( 117 to -80) with respect to the +1 transcription start site. E2F1-mediated suppression of LTR activity was observed in a wide range of human cell lines. Expression of E2F1 by a transgene showed an inhibitory effect on the levels of reverse transcriptase activity obtained upon introduction of the proviral genome into cells. The data presented in this study suggest that cellular regulatory proteins involved in the progression of cells through the mitotic cycle could play crucial roles in determining the efficiency of HIV-1 replication during the various stages of infection. The possible roles of these factors in viral latency and activation are discussed. PMID- 7474113 TI - The inhibition of cultured myoblast differentiation by the simian virus 40 large T antigen occurs after myogenin expression and Rb up-regulation and is not exerted by transformation-competent cytoplasmic mutants. AB - We have investigated the mechanism by which the simian virus 40 large T antigen (SVLT) interferes with the differentiation of C2 myoblasts. SVLT mutants, defective either in the Rb binding site, near the N-terminal end, in a region that affects binding to p53, or in the nuclear transport signal, were also employed to determine whether the interference was especially dependent on these functional domains. It was found that wild-type (wt) SVLT strongly inhibited the terminal differentiation of mouse C2 myoblasts, but this arrest occurred only after the synthesis of myogenin, an initial step in biochemical differentiation. Neither the synthesis nor some basic activities of MyoD appeared to be affected by wt SVLT. In these transformants, mitogen depletion elicited an increase in the Rb level comparable to that in normal C2 cells; wt SVLT, however, promoted the phosphorylation of a large part of the induced Rb. Mutations affecting nuclear transport were far more critical for the ability to interfere with myogenic differentiation than were those affecting the transforming potential; cytoplasmic SVLT expression was fully compatible with the terminal differentiation of C2 cells, despite enabling them to grow in semisolid medium, thus showing that the myogenesis-inhibiting property can be dissociated from transforming competence. The remaining SVLT mutants presented different degrees of ability to inhibit differentiation (as shown by the expression of tissue-specific markers in transformants). The inhibiting mutants, including the Rb binding site mutant, were able to promote a higher state of Rb phosphorylation than that observed in either normal cells or cytoplasmic-SVLT transformants. PMID- 7474114 TI - Chromosome mapping of Rmp-4, a gonad-dependent gene encoding host resistance to mousepox. AB - DBA/2 (D2) mice are susceptible and C57BL/6 (B6) mice are resistant to lethal mousepox. A congenic resistant strain, D2.B6-Rmp-4r (D2.R4), was developed by serially backcrossing male mice that survived ectromelia virus infection with D2 mice, beginning with (B6 x D2)F1 mice. Male D2.R4 mice were at least 300-fold more resistant to lethal mousepox than male D2 mice. Female D2.R4 mice were 100 fold more resistant than male D2.R4 mice and 500-fold more resistant than female D2 mice. Neonatal gonadectomy prevented development of resistance in D2.R4 mice of both sexes. Differences in resistance between strains and between sexes correlated with restriction of virus replication in spleen and liver, but gender differences were less evident in liver than in spleen. High-resolution interval mapping of the 19 autosomes of D2.R4 mice using dispersed informative microsatellites as marker loci revealed a segment of distal chromosome 1 to be of B6 origin. Haplotypes for a marker locus, D1Mit57, from the differential segment were determined in (D2.R4 x D2)F1 x D2 backcross mice, which were then infected with ectromelia virus. Significantly more heterozygotes than homozygotes survived ectromelia virus infection in both sexes. Whereas nearly all surviving males were heterozygotes, 44% of surviving females were homozygotes. These results indicate that resistance in D2.R4 mice is determined by a gonad-dependent gene on distal chromosome 1, provisionally named Rmp-4, and by an ovary-dependent factor that is not genetically linked to Rmp-4. PMID- 7474115 TI - Mutations that specifically impair the DNA binding activity of the herpes simplex virus protein UL42. AB - The herpes simplex virus DNA polymerase is a heterodimer consisting of a catalytic subunit and the protein UL42, which functions as a processivity factor. It has been hypothesized that UL42 tethers the catalytic subunit to the DNA template by virtue of DNA binding activity (J. Gottlieb, A. I. Marcy, D. M. Coen, and M. D. Challberg, J. Virol. 64:5976-5987, 1990). Relevant to this hypothesis, we identified two linker insertion mutants of UL42 that were unable to bind to a double-stranded-DNA-cellulose column but retained their ability to bind the catalytic subunit. These mutants were severely impaired in the stimulation of long-chain-DNA synthesis by the catalytic subunit in vitro. In transfected cells, the expressed mutant proteins localized to the nucleus but were nonetheless deficient in complementing the growth of a UL42 null virus. Thus, unlike many other processivity factors, UL42 appears to require an intrinsic DNA binding activity for its function both in vitro and in infected cells. Possible mechanisms for the activity of UL42 and its potential as a drug target are discussed. PMID- 7474116 TI - Differences in the capacity of reovirus strains to induce apoptosis are determined by the viral attachment protein sigma 1. AB - Reoviruses are important models for studies of viral pathogenesis; however, the mechanisms by which these viruses produce cytopathic effects in infected cells have not been defined. In this report, we show that murine L929 (L) cells infected with prototype reovirus strains type 1 Lang (TIL) and type 3 Dearing (T3D) undergo apoptosis and that T3D induces apoptosis to a substantially greater extent than T1L. Using T1L x T3D reassortant viruses, we found that differences in the capacity of T1L and T3D to induce apoptosis are determined by the viral S1 gene segment, which encodes the viral attachment protein sigma 1 and the non virion-associated protein sigma 1s. Apoptosis was induced by UV-inactivated, replication-incompetent reovirus virions, which do not contain sigma 1s and do not mediate its synthesis in infected cells. Additionally, T3D-induced apoptosis was inhibited by anti-reovirus monoclonal antibodies that inhibit T3D cell attachment and disassembly. These results indicate that sigma 1, rather than sigma 1s, is required for induction of apoptosis by the reovirus and suggest that interaction of virions with cell surface receptors is an essential step in this mechanism of cell killing. PMID- 7474117 TI - Isolation and characterization of a new simian T-cell leukemia virus type 1 from naturally infected celebes macaques (Macaca tonkeana): complete nucleotide sequence and phylogenetic relationship with the Australo-Melanesian human T-cell leukemia virus type 1. AB - A study of simian T-cell leukemia virus type 1 (STLV-1) infection in a captive colony of 23 Macaca tonkeana macaques indicated that 17 animals had high human T cell leukemia virus type 1 (HTLV-1) antibody titers. Genealogical analysis suggested mainly a mother-to-offspring transmission of this STLV-1. Three long term T-cell lines, established from peripheral blood mononuclear cell cultures from three STLV-1-seropositive monkeys, produced HTLV-1 Gag and Env antigens and retroviral particles. The first complete nucleotide sequence of an STLV-1 (9,025 bp), obtained for one of these isolates, indicated an overall genetic organization similar to that of HTLV-1 but with a nucleotide variability for the structural genes ranging from 7.8 to 13.1% compared with the HTLV-1 ATK and STLV 1 PTM3 Asian prototypes. The Tax and Rex regulatory proteins were well conserved, while the pX region, known to encode new proteins in HTLV-1 (open reading frames I and II), was more divergent than that in the ATK strain. Furthermore, a fragment of 522 bp of the gp21 env gene from uncultured peripheral blood mononuclear cell DNAs from five of the STLV-1-infected monkeys was sequenced. Phylogenetic trees constructed with the long terminal repeat and env (gp46 and gp21) regions demonstrated that this new STLV-1 occupies a unique position within the Asian STLV-1 and HTLV-1 isolates, being, by most analyses, related more to the Australo-Melanesian HTLV-1 topotype than to any other Asian STLV-1. These data raise new hypotheses on the possible interspecies viral transmission between monkeys carrying STLV-1 and early Australoid settlers, ancestors of the present day Australo-Melanesian inhabitants, during their migrations from the Southeast Asian land mass to the greater Australian continent. PMID- 7474118 TI - Retrovirus infection: effect of time and target cell number. AB - Using a model amphotropic recombinant retrovirus encoding the Escherichia coli lacZ gene and quantitative assays to measure virus infection, we have determined the effects of time and target cell number on infectivity. Infection of various numbers of NIH 3T3 fibroblasts showed that the extent of lacZ virus infection was dependent on virus concentration and independent of target cell number. These results demonstrate that multiplicity of infection is not an accurate predictor of the efficiency of retroviral infection. Varying the time of viral infection revealed that maximal infection occurred after greater than 24 h of exposure of the cells to the lacZ virus. Half-maximal infection occurred after 5 h of exposure. After 2 h of adsorption at 37 degrees C, the majority of infectious virus was not adsorbed to cells but was unbound and able to infect other cells. These results are discussed in terms of both their relevance to the fundamental biology of retrovirus infection and the use of recombinant retroviruses for retrovirus-mediated gene transfer with purposes of gene therapy. PMID- 7474119 TI - Inhibitory effect of nitric oxide on the replication of a murine retrovirus in vitro and in vivo. AB - Nitric oxide (NO) exerts microbicidal effects on a broad spectrum of pathogens, including viruses, but its antiretrovirus properties have not yet been described. The purpose of this study was to determine whether NO inhibits murine Friend leukemia virus (FV) replication in vitro and to what extent NO may play a role in defenses against FV infection in mice. Three NO-generating compounds were studied: 3-morpholino-sydononimine (SIN-1), sodium nitroprusside (SNP), and S nitroso-N-acetylpenicillamine (SNAP). The effects of these three compounds were compared with those of their controls (SIN-1C, potassium ferricyanide, and N acetylpenicillamine, respectively), which do not generate NO and with that of sodium nitrite (NaNO2). SIN-1, SNP, and SNAP inhibited FV replication in dunni cells in a concentration-dependent manner. In contrast, no significant inhibitory effect was observed with the three controls or NaNO2. Furthermore, the addition of superoxide dismutase did not alter the inhibitory effect of SIN-1, which is also known to generate superoxide anions. No dunni cell toxicity was observed in the range of concentrations tested. We also assessed the effect of NO produced by activated macrophages on FV replication. Macrophages activated by gamma interferon and lipopolysaccharide inhibited FV replication in a concentration dependent manner. This inhibition was due in part to NO production, since it was reversed by NG-monomethyl L-arginine, a competitive inhibitor of NO synthase. In vivo administration of NG-nitro-L-arginine methyl ester, a competitive inhibitor of NO synthase, significantly increased the viral load in spleen cells of FV infected mice. These results suggested that NO may play a role in defenses against the murine Friend leukemia retrovirus. PMID- 7474121 TI - Postoligomerization folding of human cytomegalovirus glycoprotein B: identification of folding intermediates and importance of disulfide bonding. AB - Human cytomegalovirus glycoprotein B (gB or UL55) has been demonstrated to be a disulfide-linked homodimer within the envelope of mature virions. Previously, it has been shown that gB undergoes a rapid dimerization nearly coincident with its synthesis. Following dimerization, the molecule slowly folds into a form which can be transported from the endoplasmic reticulum. In this study we have examined the prolonged folding of gB by using a set of defined gB-reactive murine monoclonal antibodies and gB expressed as a recombinant protein in the absence of other human cytomegalovirus proteins. Our results have documented a folding pathway consistent with the relatively rapid dimerization of the translation product followed by delayed conversion into a fully folded molecule. Assembly of the dominant antigenic domain of gB, AD-1, preceded dimerization and folding of the molecule. The fully folded dimer was heat stable, but its conformation was altered by treatment with 2% sodium dodecyl sulfate (SDS), whereas an oligomeric folding intermediate was both heat and SDS stable. Postoligomerization disulfide bond formation could be demonstrated during folding of gB, suggesting that the formation of these covalent bonds could contribute to the prolonged folding of this glycoprotein. PMID- 7474120 TI - Effects of anti-E2 monoclonal antibody on sindbis virus replication in AT3 cells expressing bcl-2. AB - Antibodies directed to Sindbis virus (SV) envelope protein E2 are able to control virus replication in vivo and in persistently infected cultures of neurons in vitro. We investigated the mechanisms by which anti-E2 monoclonal antibody (MAb) alters virus replication by using AT3 rat prostatic carcinoma cells expressing the inhibitor of apoptosis bcl-2. Treatment of SV-infected AT3-bcl-2 cells with anti-E2 MAb G5 for 2 h decreased the rate of virus release for 6 to 8 h after removal of the antibody. Electron microscopic analysis of MAb-treated cells revealed that failure of virus release was linked to a defect in the budding process. The decrease in extracellular virus particles occurred despite continued formation of nucleocapsids and synthesis of envelope glycoproteins. MAb treatment delayed the inhibition of K+ influx and shutoff of host cell protein synthesis by SV infection in a dose-dependent manner. Synthesis of host cell factors and of nonstructural polyprotein precursors required for the formation of initial replication complexes was also prolonged, causing a slower shutdown of overall viral RNA synthesis. We conclude that one mechanism by which anti-E2 MAb treatment down-regulates SV replication is by reestablishing certain critical host cell functions in infected cells. PMID- 7474123 TI - Mutagenic analysis of human immunodeficiency virus type 1 Vpr: role of a predicted N-terminal alpha-helical structure in Vpr nuclear localization and virion incorporation. AB - The Vpr gene product of human immunodeficiency virus type 1 is a virion associated protein that is important for efficient viral replication in nondividing cells such as macrophages. At the cellular level, Vpr is primarily localized in the nucleus when expressed in the absence of other viral proteins. Incorporation of Vpr into viral particles requires a determinant within the p6 domain of the Gag precursor polyprotein Pr55gag. In the present study, we have used site-directed mutagenesis to identify a domain(s) of Vpr involved in virion incorporation and nuclear localization. Truncations of the carboxyl (C)-terminal domain, rich in basic residues, resulted in a less stable Vpr protein and in the impairment of both virion incorporation and nuclear localization. However, introduction of individual substitution mutations in this region did not impair Vpr nuclear localization and virion incorporation, suggesting that this region is necessary for the stability and/or optimal protein conformation relevant to these Vpr functions. In contrast, the substitution mutations within the amino (N) terminal region of Vpr that is predicted to adopt an alpha-helical structure (extending from amino acids 16 to 34) impaired both virion incorporation and nuclear localization, suggesting that this structure may play a pivotal role in modulating both of these biological properties. These results are in agreement with a recent study showing that the introduction of proline residues in this predicted alpha-helical region abolished Vpr virion incorporation, presumably by disrupting this secondary structure (S. Mahalingam, S. A. Khan, R. Murali, M. A. Jabbar, C. E. Monken, R. G. Collman, and A. Srinivasan, Proc. Natl. Acad. Sci. USA 92:3794-3798, 1995). Interestingly, our results show that two Vpr mutants harboring single amino acid substitutions (L to F at position 23 [L23F] and A30F) on the hydrophobic face of the predicted helix coded for relatively stable proteins that retained their ability to translocate to the nucleus but exhibited dramatic reduction in Vpr incorporation, suggesting that this hydrophobic face might mediate protein-protein interactions required for Vpr virion incorporation but not nuclear localization. Furthermore, a single mutation (E25K) located on the hydrophilic face of this predicted alpha-helical structure affected not only virion incorporation but also nuclear localization of Vpr. The differential impairment of Vpr nuclear localization and virion incorporation by mutations in the predicted N-terminal alpha-helical region suggests that this region of Vpr plays a role in both of these biological functions of Vpr. PMID- 7474122 TI - Translation of the human papillomavirus type 16 E7 oncoprotein from bicistronic mRNA is independent of splicing events within the E6 open reading frame. AB - In this study we investigated the translational capacities of bicistronic and spliced mRNAs originating from the E6 and E7 regions of the high-risk genital human papillomavirus type 16 (HPV-16) and the low-risk HPV-11. For HPV-16 it was found, unexpectedly, that E7 protein could be translated from full-length bicistronic E6-E7 mRNAs. E6*I and E6*II splicing events were not required for E7 synthesis, nor did splicing increase the efficiency of E7 translation significantly. In cells, E7 synthesis from all known naturally occurring mRNA structures was very inefficient compared with that from synthetic monocistronic controls, suggesting that HPV-16 employs translational mechanisms to restrict E7 protein levels. For HPV-11, only RNAs initiated at the P264 promoter, located within the E6 open reading frame, were capable of providing an efficient template for E7 synthesis. P264-initiated mRNAs were as efficient in vivo as monocistronic controls, suggesting that the low-risk HPV-11 does not limit E7 synthesis by translational mechanisms. A detailed analysis of HPV-16 templates by using site directed mutagenesis showed that the majority of ribosomes which ultimately translate E7 have not reinitiated after translating some or all of the upstream open reading frames. The data support a model in which the failure of 40S ribosomal initiation complexes to recognize the E6 AUG renders them capable of proceeding efficiently to translate E7. PMID- 7474124 TI - Differences in the role of the cytoplasmic domain of human parainfluenza virus fusion proteins. AB - We have investigated the roles of the cytoplasmic domains of the human parainfluenza virus type 2 (PI2) and type 3 (PI3) fusion (F) proteins in protein transport and cell fusion activity. By using the vaccinia virus-T7 transient expression system, a series of F protein cytoplasmic tail truncation mutants was studied with respect to intracellular and surface expression and the ability to induce cell fusion when coexpressed with the corresponding hemagglutinin neuraminidase (HN) proteins. All of the cytoplasmic tail truncation mutants of PI2F were expressed at high levels intracellularly or on cell surfaces as measured by immunoprecipitation and cell surface biotinylation assays. In addition, when coexpressed with PI2HN, these truncation mutants of PI2F were all found to be essentially unimpaired in the ability to induce cell fusion as measured by a quantitative cell fusion assay. In contrast, surface expression and cell fusion activity were found to be eliminated by a mutant of PI3F in which the entire cytoplasmic tail was deleted, and the mutant protein appeared to be unable to assemble into a high-molecular-weight oligomeric structure. To further investigate whether there is a specific sequence requirement in the cytoplasmic tail of PI3F, a chimeric protein consisting of the PI3F extracellular and transmembrane domains and the PI2F cytoplasmic tail was constructed. This chimeric protein was detected on the surface, and it was capable of inducing cell fusion when expressed together with PI3HN, although the fusogenic activity was reduced compared with that of wild-type PI3F. These results demonstrate that although PI2 and PI3 viruses belong to the same parainfluenza virus genus, these viruses show marked differences with respect to functional requirements for the cytoplasmic tail of the F glycoprotein. PMID- 7474125 TI - A transcript from the long terminal repeats of a murine retrovirus associated with trans activation of cellular genes. AB - Infection of human or murine cells with murine leukemia viruses rapidly increases the expression of a number of genes that belong to the immunoglobulin superfamily and are involved in T-lymphocyte activation, including the class I major histocompatibility complex antigens. We have reported recently that the long terminal repeat (LTR) of Moloney murine leukemia virus encodes a trans activator which induces transcription and expression of class I major histocompatibility complex genes and certain cytokine genes. The portion of the LTR responsible for trans activation was mapped by deletions to lie within the U3 region. We demonstrate here that a transcript is initiated within the U3 region and that its presence correlates with the trans-activating activity. Analysis of the LTR region reveals a potential internal promoter element for RNA polymerase III transcription within the U3 region. Studies with polymerase inhibitors suggest that this LTR transcript, designated let (LTR-encoded trans activator), is a product of RNA polymerase III. The mechanisms whereby RNA leukemia viruses cause lymphoid neoplasia after a long latent period have been extensively studied but are only partially understood. The region of the LTR identified here as being important in trans activation has recently been shown to be a critical determinant of the leukemogenicity and latency of Moloney murine leukemia virus. These findings suggest a novel mechanism of retrovirus-induced activation of cellular gene expression, potentially contributing to leukemogenesis. PMID- 7474127 TI - Characterization of ts 16, a temperature-sensitive mutant of vaccinia virus. AB - We have characterized a temperature-sensitive mutant of vaccinia virus, ts16, originally isolated by Condit et al. (Virology 128:429-443, 1983), at the permissive and nonpermissive temperatures. In a previous study by Kane and Shuman (J. Virol 67:2689-2698, 1993), the mutation of ts16 was mapped to the I7 gene, encoding a 47-kDa protein that shows partial homology to the type II topoisomerase of Saccharomyces cerevisiae. The present study extends previous electron microscopy analysis, showing that in BSC40 cells infected with ts16 at the restrictive temperature (40 degrees C), the assembly was arrested at a stage between the spherical immature virus and the intracellular mature virus (IMV). In thawed cryosections, a number of the major proteins normally found in the IMV were subsequently localized to these mutant particles. By using sucrose density gradients, the ts16 particles were purified from cells infected at the permissive and nonpermissive temperatures. These were analyzed by immunogold labelling and negative-staining electron microscopy, and their protein composition was determined by sodium dodecyl sulfate-polyacrylamide gel electrophoresis. While the ts16 virus particles made at the permissive temperature appeared to have a protein pattern identical to that of wild-type IMV, in the mutant particles the three core proteins, p4a, p4b, and 28K, were not proteolytically processed. Consistent with previous data the sucrose-purified particles could be labelled with [3H]thymidine. In addition, anti-DNA labelling on thawed cryosections suggested that most of the mutant particles had taken up DNA. On thawed cryosections of cells infected at the permissive temperature, antibodies to I7 labelled the virus factories, the immature viruses, and the IMVs, while under restrictive conditions these structures were labelled much less, if at all. Surprisingly, however, by Western blotting (immunoblotting) the I7 protein was present in similar amounts in the defective particles and in the IMVs isolated at the permissive temperature. Finally, our data suggest that at the nonpermissive temperature the assembly of ts16 is irreversibly arrested in a stage at which the DNA is in the process of entering but before the particle has completely sealed, as monitored by protease experiments. PMID- 7474126 TI - Persistent infection of macaques with simian-human immunodeficiency viruses. AB - Chimeric simian-human immunodeficiency viruses (SHIV) containing the human immunodeficiency virus type 1 (HIV-1) tat, rev, env, and, in some cases, vpu genes were inoculated into eight cynomolgus monkeys. Viruses could be consistently recovered from the CD8-depleted peripheral blood lymphocytes of all eight animals for at least 2 months. After this time, virus isolation varied among the animals, with viruses continuing to be isolated from some animals beyond 600 days after inoculation. The level of viral RNA in plasma during acute infection and the frequency of virus isolation after the initial 2-month period were higher for the Vpu-positive viruses. All of the animals remained clinically healthy, and the absolute numbers of CD4-positive lymphocytes were stable. Antibodies capable of neutralizing HIV-1 were generated at high titers in animals exhibiting the greatest consistency of virus isolation. Strain-specific HIV-1 neutralizing antibodies were initially elicited, and then more broadly neutralizing antibodies were elicited. env sequences from two viruses isolated more than a year after infection were analyzed. In the Vpu-negative SHIV, for which virus loads were lower, a small amount of env variation, which did not correspond to that found in natural HIV-1 variants, was observed. By contrast, in the Vpu-positive virus, which was consistently isolated from the host animal, extensive variation of the envelope glycoproteins in the defined variable gp120 regions was observed. Escape from neutralization by CD4 binding site monoclonal antibodies was observed for the viruses with the latter envelope glycoproteins, and the mechanism of escape appears to involve decreased binding of the antibody to the monomeric gp120 glycoproteins. The consistency with which SHIV infection of cynomolgus monkeys is initiated and the similarities in the neutralizing antibody response to SHIV and HIV-1 support the utility of this model system for the study of HIV-1 prophylaxis. PMID- 7474128 TI - Glycoproteins E and I facilitate neuron-to-neuron spread of herpes simplex virus. AB - Two herpes simplex virus (HSV) glycoproteins E and I (gE and gI) form a heterooligomer which acts as an Fc receptor and also facilitates cell-to-cell spread of virus in epithelial tissues and between certain cultured cells. By contrast, gE-gI is not required for infection of cells by extracellular virus. HSV glycoproteins gD and gJ are encoded by neighboring genes, and gD is required for both virus entry into cells and cell-to-cell spread, whereas gJ has not been shown to influence these processes. Since HSV infects neurons and apparently spreads across synaptic junctions, it was of interest to determine whether gD, gE, gI and gJ are also important for interneuronal transfer of virus. We tested the roles of these glycoproteins in neuron-to-neuron transmission of HSV type 1 (HSV-1) by injecting mutant viruses unable to express these glycoproteins into the vitreous body of the rat eye. The spread of virus infection was measured in neuron-rich layers of the retina and in the major retinorecipient areas of the brain. Wild-type HSV-1 and a gJ- mutant spread rapidly between synaptically linked retinal neurons and efficiently infected major retinorecipient areas of the brain. gD mutants, derived from complementing cells, infected only a few neurons and did not spread in the retina or brain. Mutants unable to express gE or gI were markedly restricted in their ability to spread within the retina, produced 10-fold-less virus in the retina, and spread inefficiently to the brain. Furthermore, when compared with wild-type HSV-1, gE- and gI- mutants spread inefficiently from cell to cell in cultures of neurons derived from rat trigeminal ganglia. Together, our results suggest that the gE-gI heterooligomer is required for efficient neuron-to-neuron transmission through synaptically linked neuronal pathways. PMID- 7474129 TI - Syncytium induction in primary CD4+ T-cell lines from normal donors by human immunodeficiency virus type 1 isolates with non-syncytium-inducing genotype and phenotype in MT-2 cells. AB - Human immunodeficiency virus type 1 (HIV-1) isolates classified as syncytium inducing (SI) or non-SI (NSI) in the MT-2 T-cell line exhibit characteristic sequence differences in the V1-V2 and V3 regions of the env gene. Seven HIV-1 isolates were phenotyped as NSI or SI in the MT-2 cell line. Unexpectedly, all four NSI viruses induced large syncytia 4 to 8 days postinoculation in a panel of five primary CD4+ T-cell lines (including two clones) generated from the peripheral blood of normal donors by exposure to infectious HIV-1, inactivated HIV-1, or Epstein-Barr virus. The primary T-cell lines yielded neither HIV-1 provirus nor infectious HIV by PCR analysis or exhaustive coculture with phytohemagglutinin-treated blast cells. Three isolates (TC354, PK1, and PK2) were biologically cloned and retained their SI or NSI phenotypes in MT-2 and primary T cell lines. The biologically cloned provirus DNA was also used to clone and sequence the relevant V2 and V3 regions of the env genes. The amino acid sequences of the V2 and V3 regions were characteristic of patterns already reported for the NSI, switch NSI, and SI phenotypes, respectively. This evidence precludes the possibility that these results were due to contamination of the NSI isolates with SI virus. The results unequivocally indicate that HIV-1 isolates with the NSI genotype and phenotype in MT-2 cells may actively induce syncytia in cloned CD4+ T cells in vitro and support the view that direct cytopathic effects may contribute to the steady decline in CD4+ T cells in asymptomatic HIV-1 seropositive patients without detectable SI virus. PMID- 7474130 TI - Interaction between duck hepatitis B virus and a 170-kilodalton cellular protein is mediated through a neutralizing epitope of the pre-S region and occurs during viral infection. AB - Identification of cell surface viral binding proteins is important for understanding viral attachment and internalization. We have fused the pre-S domain of the duck hepatitis B virus (DHBV) large envelope protein to glutathione S-transferase and demonstrated a 170-kDa binding protein (p170) in [35S]methionine-labeled duck hepatocyte lysates. This glycoprotein was found abundantly in all extrahepatic tissues infectible with DHBV and in some noninfectible tissues, though it is not secreted into the blood. The interaction of pre-S fusion protein with p170 was competitively inhibited by wild-type DHBV in a dose-dependent manner. In addition, infection of hepatocytes with DHBV blocked the binding of pre-S fusion protein to p170, which suggests a biological role for p170 during natural infection. The p170 binding site was mapped to a conserved sequence of 16 amino acid residues (positions 87 to 102) by using 24 pre-S deletion mutants; this binding domain coincides with a major virus neutralizing antibody epitope. Furthermore, site-directed mutagenesis revealed that an arginine residue at position 97 is critical for p170 binding. p170 was purified by a combination of ion-exchange and affinity chromatographies, and four peptide sequences were obtained. Two peptides showed significant similarities to human and animal carboxypeptides H, M, and N. Taken together, these results raise the possibility that the p170 binding protein is important during the replication cycle of DHBV. PMID- 7474131 TI - Release of the catalytic domain N(o) from the herpes simplex virus type 1 protease is required for viral growth. AB - The herpes simplex virus type 1 (HSV-1) protease and its substrate, ICP35, are involved in the assembly of viral capsids and required for efficient viral growth. The full-length protease (Pra) consists of 635 amino acid (aa) residues and is autoproteolytically processed at the release (R) site and the maturation (M) site, releasing the catalytic domain No (VP24), Nb (VP21), and a 25-aa peptide. To understand the biological importance of cleavage at these sites, we constructed several mutations in the cloned protease gene. Transfection assays were performed to determine the functional properties of these mutant proteins by their abilities to complement the growth of the protease deletion mutant m100. Our results indicate that (i) expression of full-length protease is not required for viral replication, since a 514-aa protease molecule lacking the M site could support viral growth; and that (ii) elimination of the R site by changing the residue Ala-247 to Ser abolished viral replication. To better understand the functions that are mediated by proteolytic processing at the R site of the protease, we engineered an HSV-1 recombinant virus containing a mutation at this site. Analysis of the mutant A247S virus demonstrated that (i) the mutant protease retained the ability to cleave at the M site and to trans process ICP35 but failed to support viral growth on Vero cells, demonstrating that release of the catalytic domain No from Pra is required for viral replication; and that (ii) only empty capsid structures were observed by electron microscopy in thin sections of A247S-infected Vero cells, indicating that viral DNA was not encapsidated. Our results demonstrate that processing of ICP35 is not sufficient to support viral replication and provide genetic evidence that the HSV-1 protease has nuclear functions other than enzymatic activity. PMID- 7474132 TI - Processing of the envelope glycoprotein gp160 in immunotoxin-resistant cell lines chronically infected with human immunodeficiency virus type 1. AB - We describe the isolation and characterization of variant cell lines which are chronically infected with the human immunodeficiency virus (HIV) and resistant to the action of immunotoxins directed against the HIV envelope protein. These variants all produce normal levels of HIV proteins, budding virions, and the envelope protein precursor gp160. Two of the variants, 10E and 11E, contain a mutation within the env gene which results in the production of a truncated precursor and altered processing and transport of the protein to the cell surface. Variants B9 and G4 are defective in gp160 cleavage and do not efficiently transport the envelope protein to the cell surface. There are no mutations in the expressed viruses of B9 and G4. These cell lines express higher levels of CD4 protein and mRNA than H9/NL4-3. Thus, 10E, 11E, B9, and G4 have escaped immunotoxin action by downmodulating the envelope protein from their cell surfaces. None of these variants produce infectious HIV. Two other immunotoxin resistant variants, E9-3 and 41-17, produce normal levels of gp160, efficiently transport the cleaved and processed subunits to the cell surface, and secrete infectious HIV. These studies identify alterations in gp160 processing that underscore the importance of the relationship between HIV and the cell that it infects. PMID- 7474133 TI - Evi-5, a common site of retroviral integration in AKXD T-cell lymphomas, maps near Gfi-1 on mouse chromosome 5. AB - We have identified a novel common site of retroviral integration, Evi-5, in AKXD T-cell lymphomas. All proviruses located at Evi-5 are clustered within a 7-kb genomic region and, where determined, are oriented in the same transcriptional direction. Interspecific backcross analysis localized Evi-5 to mouse chromosome 5, where it cosegregated with another common viral integration site, Gfi-1. Gfi-1 encodes a novel zinc finger transcription factor whose expression is thought to be important for interleukin-2 signaling. Physical mapping studies showed that Evi-5 is located approximately 18 kb upstream of Gfi-1, and Southern analysis showed that Gfi-1, like Evi-5, is a common integration site in AKXD T-cell tumors. With one exception, Evi-5 and Gfi-1 integrations were mutually exclusive. Ten of the tumors with Evi-5 or Gfi-1 integrations also harbored viral integrations at other common integration sites causally associated with T-cell disease. These results are consistent with the hypothesis that T-cell lymphomagenesis is a multistep disease and that viral integration at Evi-5 or Gfi 1 is causally associated with this disease process. PMID- 7474135 TI - Emergence of virus escape mutants after immunization with epitope vaccine. AB - BALB/c and C57BL/6J mice were immunized with recombinant vaccines consisting of lymphocytic choriomeningitis virus CD8+ T-lymphocyte epitopes and a carrier protein. During challenge infection with WE strain lymphocytic choriomeningitis virus, mutants with alterations in distinct amino acid residues of the epitopic nonapeptides appeared and multiplied. Splenocytes from WE-infected BALB/c mice lysed cells coated with the WE-type epitope; lysis was considerably less effective when the epitopic nonapeptide with which the syngeneic cells had been sensitized was the mutated form. Neither target was lysed by splenocytes from BALB/c mice infected with the variant virus. Mutants were not detected in F1 hybrid mice immunized with two viral epitopes that were restricted by class I molecules of both parents. PMID- 7474134 TI - Frequent disruption of the Nf1 gene by a novel murine AIDS virus-related provirus in BXH-2 murine myeloid lymphomas. AB - Evi-2, a common site of viral integration in BXH-2 myeloid lymphomas, is located within a large intron of the Nf1 tumor suppressor gene. Viral integration at Evi 2 appears to induce disease by disrupting normal Nf1 expression. During our attempts to characterize the nature of the proviruses located at Evi-2, we found that approximately half of the proviruses were defective nonecotropic proviruses (A. M. Buchberg, H. G. Bedigian, N. A. Jenkins, and N. G. Copeland, Mol. Cell. Biol. 10:4658-4666, 1990). This was surprising, since most proviruses characterized at other BXH-2 common integration sites are full-length ecotropic viruses. In the studies described here, we found that this defective provirus carries two large deletions, one in pol and one in env, and is structurally related to another murine retrovirus, the murine AIDS retrovirus. By using oligonucleotide probes specific for this defective provirus, designated MRV, we showed that MRV-related proviruses are carried as endogenous germ line proviruses in most inbred strains. In addition, we identified the endogenous MRV provirus that gives rise to the defective proviruses identified at Evi-2. We present a model that accounts for the positive selection of MRV proviruses at Evi-2, which may allow selective identification of common viral integration sites harboring tumor suppressor genes. PMID- 7474136 TI - Protective efficacy of nonneutralizing monoclonal antibodies in acute infection with murine leukemia virus. AB - We have used an experimental retrovirus infection to study the roles played by different antibodies in resistance to both infection and disease. A molecularly cloned chimeric murine leukemia virus was used to induce acute lethal neurological disease in neonatal mice. A panel of monoclonal antibodies directed against the Gag and Env proteins was tested for protective efficacy. In vitro neutralization assays demonstrated that anti-Env antibodies gave different degrees of neutralization, while no anti-Gag neutralized the virus. In vivo experimental endpoints were onset of clinical signs and premoribund condition. As expected, different anti-Env antibodies demonstrated different degrees of protection which correlated with their neutralizing abilities. Surprisingly, anti Gag antibodies directed against both p15 (MA protein) and p30 (CA protein) were also protective, significantly delaying the onset of disease. No protection was seen with either of two control antibodies. The protection with anti-Gag was dose related and time dependent and was also produced with Fab fragments. Treatment with anti-Gag did not prevent viremia but resulted in a slight slowing in viremia kinetics and decreased levels of virus in the central nervous systems of mice protected from disease. These data indicate that nonneutralizing antiretroviral antibodies can influence the outcome of retroviral disease. The data also suggest a functional role for cell surface expression of Gag proteins on murine leukemia virus-infected cells. PMID- 7474137 TI - 3C-like protease of rabbit hemorrhagic disease virus: identification of cleavage sites in the ORF1 polyprotein and analysis of cleavage specificity. AB - Rabbit hemorrhagic disease virus, a positive-stranded RNA virus of the family Caliciviridae, encodes a trypsin-like cysteine protease as part of a large polyprotein. Upon expression in Escherichia coli, the protease releases itself from larger precursors by proteolytic cleavages at its N and C termini. Both cleavage sites were determined by N-terminal sequence analysis of the cleavage products. Cleavage at the N terminus of the protease occurred with high efficiency at an EG dipeptide at positions 1108 and 1109. Cleavage at the C terminus of the protease occurred with low efficiency at an ET dipeptide at positions 1251 and 1252. To study the cleavage specificity of the protease, amino acid substitutions were introduced at the P2, P1, and P1' positions at the cleavage site at the N-terminal boundary of the protease. This analysis showed that the amino acid at the P1 position is the most important determinant for substrate recognition. Only glutamic acid, glutamine, and aspartic acid were tolerated at this position. At the P1' position, glycine, serine, and alanine were the preferred substrates of the protease, but a number of amino acids with larger side chains were also tolerated. Substitutions at the P2 position had only little effect on the cleavage efficiency. Cell-free expression of the C-terminal half of the ORF1 polyprotein showed that the protease catalyzes cleavage at the junction of the RNA polymerase and the capsid protein. An EG dipeptide at positions 1767 and 1768 was identified as the putative cleavage site. Our data show that rabbit hemorrhagic disease virus encodes a trypsin-like cysteine protease that is similar to 3C proteases with regard to function and specificity but is more similar to 2A proteases with regard to size. PMID- 7474139 TI - An active-site mutation in the human immunodeficiency virus type 1 proteinase (PR) causes reduced PR activity and loss of PR-mediated cytotoxicity without apparent effect on virus maturation and infectivity. AB - Infectious retrovirus particles are derived from structural polyproteins which are cleaved by the viral proteinase (PR) during virion morphogenesis. Besides cleaving viral polyproteins, which is essential for infectivity, PR of human immunodeficiency virus (HIV) also cleaves cellular proteins and PR expression causes a pronounced cytotoxic effect. Retroviral PRs are aspartic proteases and contain two copies of the triplet Asp-Thr-Gly in the active center with the threonine adjacent to the catalytic aspartic acid presumed to have an important structural role. We have changed this threonine in HIV type 1 PR to a serine. The purified mutant enzyme had an approximately 5- to 10-fold lower activity against HIV type 1 polyprotein and peptide substrates compared with the wild-type enzyme. It did not induce toxicity on bacterial expression and yielded significantly reduced cleavage of cytoskeletal proteins in vitro. Cleavage of vimentin in mutant-infected T-cell lines was also markedly reduced. Mutant virus did, however, elicit productive infection of several T-cell lines and of primary human lymphocytes with no significant difference in polyprotein cleavage and with similar infection kinetics and titer compared with wild-type virus. The discrepancy between reduced processing in vitro and normal virion maturation can be explained by the observation that reduced activity was due to an increase in Km which may not be relevant at the high substrate concentration in the virus particle. This mutation enables us therefore to dissociate the essential function of PR in viral maturation from its cytotoxic effect. PMID- 7474138 TI - Poliovirus protein 3AB forms a complex with and stimulates the activity of the viral RNA polymerase, 3Dpol. AB - Poliovirus protein 3B (also known as VPg) is covalently linked to the 5' ends of both genomic and antigenomic viral RNA. Genetic and biochemical studies have implicated protein 3AB, the membrane-bound precursor to VPg, in the initiation of genomic RNA synthesis. We have purified 3AB to near homogeneity following thrombin cleavage of purified glutathione S-transferase-3AB. When added to transcription reaction mixtures catalyzed by poliovirus RNA polymerase (3Dpol), 3AB stimulated RNA synthesis up to 75-fold with oligo(U)-primed virion RNA, globin mRNA, and unprimed synthetic, full-length minus-strand viral RNA as the templates. Synthetic VPg also stimulated RNA synthesis but was only 1 to 2% as effective as 3AB on a molar basis. The increased level of transcription was not the result of enhancing the elongation rate of the polymerase. No evidence was found for uridylylation of 3AB or for covalent linkage to RNA transcription products. 3AB sedimented as a multimer in glycerol gradients. In the presence of the polymerase, the sedimentation rate of both proteins increased, suggesting the formation of a complex. Detergent prevented both multimerization and complex formation. The polymerase also bound to immobilized glutathione S-transferase 3AB; this procedure was used to purify the polymerase to near homogeneity. These results suggest a mechanism for bringing together 3AB, 3Dpol (or its precursor 3CD), and viral RNA in host cell membranous vesicles in which all viral RNA synthesis occurs. PMID- 7474141 TI - Biological activity of human immunodeficiency virus type 1 Vif requires membrane targeting by C-terminal basic domains. AB - Human immunodeficiency virus type 1 (HIV-1) encodes a Vif protein which is important for virus replication and infectivity. Vif is a cytoplasmic protein which exists in both membrane-associated and soluble forms. The membrane associated form is an extrinsic membrane protein which is tightly associated with the cytoplasmic side of membranes. We have analyzed the mechanism of membrane targeting of Vif and its role in HIV-1 replication. Mutagenesis studies demonstrate that C-terminal basic domains are required for membrane association. Vif mutations which disrupt membrane association also inhibit HIV-1 replication, indicating that membrane localization of Vif is likely to be required for its biological activity in vivo. Membrane binding of Vif is almost completely abolished by trypsin treatment of membranes. These results demonstrate that membrane localization of Vif requires C-terminal basic domains and interaction with a membrane-associated protein(s). This interaction may serve to direct Vif to a specific cellular site, since immunofluorescence staining and plasma membrane fractionation studies show that Vif is localized predominantly to an internal cytoplasmic compartment rather than to the plasma membrane. The mechanism of membrane targeting of Vif is different in some respects from that of other extrinsic membrane proteins, such as Ras, Src, and MARCKS, which utilize a basic domain together with a lipid modification for membrane targeting. Membrane targeting of Vif is likely to play an important role in HIV-1 replication and thus may be a therapeutic target. PMID- 7474140 TI - Herpes simplex virus trans-regulatory protein ICP27 stabilizes and binds to 3' ends of labile mRNA. AB - Previous work demonstrated that a herpes simplex virus type 1 (HSV-1) immediate early function up-regulates beta interferon but not chloramphenicol acetyltransferase reporter genes driven by the strong simian virus 40 (SV40) or cytomegalovirus promoter-enhancer regions in both transient assays and stable cell lines. The different 3' mRNA stabilization and RNA-processing signals from these two reporter genes appeared to be primarily responsible for this phenomenon. We now report that the HSV-1 ICP27 itself is sufficient to stimulate both steady-state accumulation and increased half-life of beta interferon reporter gene mRNA. Furthermore, the ability to respond directly to cotransfected ICP27 can be transferred to chloramphenicol acetyltransferase reporter genes by replacement of their SV40-derived splicing and poly(A) signals with the 3' AU rich and poly(A) RNA-processing signals from the normally highly labile beta interferon and c-myc mRNA species. ICP27 expressed in bacteria bound specifically to in vitro-generated RNA from both the beta interferon and c-myc intronless AU rich 3' RNA-processing regions, but not to the SV40-derived early-region splice signal and poly(A) sequences. By site-specific mutagenesis, we also show that individual ICP27 C-terminal amino acid residues that are positionally conserved in ICP27 homologs in other herpesviruses (D-357, E-358, H-479, C-400, C-483, and C-488) are critical for trans-regulatory activity. Importantly, several of these positions match mutations that are known to be essential for the role of ICP27 in the early-to-late switch during the virus lytic cycle. Therefore, our findings support the notion that HSV ICP27 modulates gene expression posttranscriptionally in part by targeting RNA. PMID- 7474142 TI - The amphotropic and ecotropic murine leukemia virus envelope TM subunits are equivalent mediators of direct membrane fusion: implications for the role of the ecotropic envelope and receptor in syncytium formation and viral entry. AB - The murine leukemia virus (MuLV) envelope protein was examined to determine which sequences are responsible for the differences in direct membrane fusion observed with the ecotropic and amphotropic MuLV subtypes. These determinants were studied by utilizing amphotropic-ecotropic chimeric envelope proteins that have switched their host range but retain their original fusion domain (TM subunit). Fusion was tested both in rodent cells and in 293 cells bearing the human homolog of the ecotropic MuLV receptor. The results demonstrate that the amphotropic TM is able to mediate cell-to-cell fusion to an extent equivalent to that mediated by the ecotropic TM, indicating that their fusion domains are equivalent. The "murinized" human homolog of the ecotropic receptor supports syncytium formation as well as the native murine receptor. These findings suggest that interactions between the ecotropic envelope protein and conserved sequences in the ecotropic receptor are the principal determinants of syncytium formation. The relationship of the fusion phenotype to pH-dependent infection and the route of viral entry was examined by studying virions bearing the chimeric envelope proteins. Such virions appear to enter cells via a pathway that is directed by the host range determining region of their envelope rather than by sequences that confer pH dependence. Therefore, the pH dependence of infection may not reflect the initial steps in viral entry. Thus, it appears that both the syncytium phenotype and the route of viral entry are properties of the viral receptor, the amino-terminal half of the ecotropic envelope protein, or the interaction between the two. PMID- 7474144 TI - Hepatitis delta virus mutant: effect on RNA editing. AB - During the replication cycle of hepatitis delta virus (HDV), RNA editing occurs at position 1012 on the 1679-nucleotide RNA genome. This changes an A to G in the amber termination codon, UAG, of the small form of the delta antigen (delta Ag). The resultant UGG codon, tryptophan, allows the translation of a larger form of the delta Ag with a 19-amino-acid C-terminal extension. Using HDV cDNA transfected cells, we examined the editing potential of HDV RNA mutated from G to A at 1011 on the antigenome, adjacent to normal editing site at 1012. Four procedures were used to study not only the editing of the A at 1012, but also that of the new A at 1011: (i) nucleotide sequencing, (ii) a PCR-based RNA editing assay, (iii) immunoblot assays, and (iv) immunofluorescence. Five findings are reported. (i) Even after the mutation at 1011, editing still occurred at 1012. (ii) Site 1011 itself now acted as a novel RNA-editing site. (iii) Sites 1011 and 1012 were edited independently. (iv) At later times, both sites became edited, thereby allowing the synthesis of the large form of the delta Ag (delta Ag-L). (v) Via immunofluorescence, such double editing became apparent as a stochastic event, in that groups of cells arose in which the changes had taken place. Evaluation of these findings and of those from previous studies of the stability of the HDV genomic sequence (H.J. Netter et al., J. Virol. 69:1687-1692, 1995) supports both the recent reevaluation of HDV RNA editing as occurring on antigenomic RNA (Casey and Gerin, personal communication) and the interpretation that editing occurs via the RNA-modifying enzyme known as DRADA. PMID- 7474143 TI - Selective infection of human T-lymphotropic virus type 1 (HTLV-1)-infected cells by chimeric human immunodeficiency viruses containing HTLV-1 tax response elements in the long terminal repeat. AB - Previous studies have suggested that the human immunodeficiency virus long terminal repeat (HIV LTR) enhancer/promoter sequences contribute to the replication ability of HIV in different T-cell lines; mutation of these sequences can alter HIV tropism. We have utilized site-specific mutagenesis to generate variants of HIV that exhibit specific tropism for human T-lymphotropic virus type 1 (HTLV-1) Tax-expressing CD4+ T cells. The wild-type HIV LTR NF-kappa B and Sp1 sites in an infectious molecular clone of HIV type 1 were replaced with sequences derived from the 21-bp Tax response elements (TRE) from the HTLV-1 LTR to generate TRE-containing chimeric HIVs (TRE-HIVs). The TRE-HIVs exhibit selective replication and cell killing in HTLV-infected human CD4+ T cells, but not in HTLV negative T cells. Transient transfections suggested that Tax-TRE interactions could account for the observed replication specificity. The TRE-containing HIV LTRs were synergistically activated by the HIV Tat and HTLV-1 Tax transactivators. These results demonstrate that it is possible to specifically target HIV replication and cytotoxicity to HTLV-1+, CD4+ human T cells, on the basis of Tax-TRE interactions, and provide a model for the development of specific, cytotoxic, retroviral gene therapy vectors for HTLV-1-infected cells based on alterations of the LTR transcriptional regulatory elements. They also suggest that HIV Tat can cooperate with heterologous transcriptional activators, such as Tax, which act through upstream binding sites without directly binding to DNA. PMID- 7474146 TI - Avian homologs of the mammalian low-density lipoprotein receptor family bind minor receptor group human rhinovirus. AB - Avian oocyte-specific very low density lipoprotein receptor specifically binds human rhinovirus of the minor receptor group on ligand blots and in solution. The solubilized receptor protects cell against infection in a dose-dependent manner. PMID- 7474145 TI - Evidence that flavivirus NS1-NS2A cleavage is mediated by a membrane-bound host protease in the endoplasmic reticulum. AB - Previous deletion mutagenesis studies have shown that the flavivirus NS1-NS2A clevage requires the eight C-terminal residues of NS1, constituting the cleavage recognition sequence, and sequences in NS2A far downstream of the cleavage site. We now demonstrate that replacement of all of NS1 upstream of the cleavage recognition sequence with prM sequences still allows cleavage in vivo. Thus, other than the eight C-terminal residues, NS1 is dispensable for NS1-NS2A cleavage. However, deletion of the N-terminal signal sequence abrogated cleavage, suggesting that entry into the exocytic pathway is required. Cleavage in vivo was not blocked by brefeldin A, and cleavage could occur in vitro in the presence of dog pancreas microsomes, indicating that NS1-NS2A cleavage occurs in the endoplasmic reticulum. Four in-frame deletions in NS2A were cleavage defective in vitro, as were two mutants in which NS4A-NS4B sequences were substituted for NS2A, suggesting that most of NS2A is required. A series of substitution mutants were constructed in which all Asp, Cys, Glu, His, and Ser residues in NS2A were collectively replaced; all standard proteases require at least one of these residues in their active sites. No single mutant was cleavage defective, suggesting that NS2A is not a protease. Fractionation of the microsomes indicated that the lumenal contents were not required for NS1-NS2A cleavage. It seems most likely that NS1-NS2A cleavage is effected by a host membrane-bound endoplasmic reticulum-resident protease, quite possibly signalase, and that NS2A is required to present the cleavage recognition sequence in the correct conformation to the host enzyme for cleavage. PMID- 7474147 TI - Human T-lymphotropic virus type 1 in coastal natives of British Columbia: phylogenetic affinities and possible origins. AB - Human T-lymphotropic virus type 1 (HTLV-1) infection has been discovered recently in people of Amerindian descent living in coastal areas of British Columbia, Canada. DNA sequencing combined with phylogenetic analysis and restriction fragment length polymorphism (RFLP) typing of HTLV-1 strains recovered from these British Columbia Indians (BCI) was conducted. Sequence-based phylogenetic trees distributed the BCI isolates among the Japanese subcluster (subcluster B) and the geographically widely distributed subcluster (subcluster A) of the large HTLV-1 cosmopolitan cluster. Long terminal repeat (LTR) RFLP typing revealed three distinct, equally frequent LTR cleavage patterns, two of which were of previously recognized Japanese and widely dispersed cosmopolitan types. A third, new cleavage pattern was detected which may have arisen by recombination between two other HTLV-1 genotypes. Our results suggest multiple origins for HTLV-1 in BCI, which are equally consistent with (i) a cluster of recent sporadic infections, (ii) ancient endemic vertical transmission through Amerindian lineages, or (iii) both. PMID- 7474148 TI - Differential downregulation of CD46 by measles virus strains. AB - Recently, we found that several lymphotropic wild-type isolates of measles virus (MV) did not lead to the downregulation of CD46 following infection. We hypothesized that either the site of virus isolation, e.g., throat swab versus peripheral blood mononuclear cells, or the cell type used for the isolation may exert selective pressure on a mixed population of viruses, resulting in isolates with the differential properties observed. This hypothesis has been tested by simultaneously isolating MV from a throat swab and peripheral blood mononuclear cells from a single patient by cultivation on B95 and Vero cells. We report that neither the source of MV nor the cell type used for isolation directly influenced the capacity for CD46 modulation of these MV isolates. PMID- 7474149 TI - The S2 subunit of the murine coronavirus spike protein is not involved in receptor binding. AB - The receptor-binding capacity of the S2 subunit of the murine coronavirus S protein was examined by testing the inhibition of virus-receptor binding. Sp-4 virus and S1N(330), which consists of the N-terminal 330 amino acids of the S1 protein, both of which exhibited receptor-binding capacity, were able to prevent the binding of cl-2 virus to the receptor, while the mutant protein S1N(330)-159, which failed to bind to the receptor protein, and S2TM-, which lacks the transmembrane and cytoplasmic domains normally existing in the S2, were unable to prevent the binding of cl-2. By using cultured DBT cells, it was revealed that the infection of cells by cl-2 virus was significantly inhibited by S1N(330) but not by S2TM-. These results indicate that the S2 protein is not involved in the receptor binding of murine coronaviruses. PMID- 7474150 TI - Active foamy virus proteinase is essential for virus infectivity but not for formation of a Pol polyprotein. AB - To analyze proteolytic processing of foamy (spuma) retroviruses, two mutations were generated in the presumed active-site triplet Asp-Ser-Gly in the predicted proteinase (PR) region of the human foamy virus (HSRV). The mutations changed either the presumed catalytic aspartic acid residue to a catalytically incompetent alanine or the adjacent serine to a threonine found in most cellular and retroviral proteases at this position. Both mutations were cloned into the full-length infectious HSRV DNA clone. Wild-type and S/T mutant genomes directed the synthesis of particles with similar infectious titers, while the HSRV D/A PR mutant was noninfectious. Immunoblot analysis of transfected cells revealed identical patterns for the wild-type and for the S/T PR mutant. HSRV D/A mutant transfected cells expressed only a single Gag polyprotein of 78 kDa instead of the 78-kDa-74-kDa doublet found in HSRV-infected or wild-type-transfected cells. Analysis with pol-specific antisera yielded a protein of approximately 120 kDa reactive with antisera against pol- but not gag-specific domains. No Gag-Pol polyprotein was detected in this study. Electron microscopy analysis of transfected cells showed heterogeneous particle morphology in the case of the D/A mutant, with particles of normal appearance and particles of aberrant size and shape. These results indicate that foamy viruses have an aspartic PR that is essential for infectivity but not for formation of the 120-kDa Pol polyprotein. PMID- 7474151 TI - A novel V beta 2-specific endogenous mouse mammary tumor virus which is capable of producing a milk-borne exogenous virus. AB - We have previously reported new Mtv loci, Mtv-48 and -51, in the Japanese laboratory mouse strains CS and NC. Here we show by backcross analysis that both Mtv-48 and -51 cosegregate with very slow deletion of T cells bearing V beta 2. The nucleotide sequences of the open reading frames in the 3' long terminal repeats of Mtv-48 and -51 were very similar to those of Mtv-DDO, mouse mammary tumor virus C4 [MMTV(C4)], and MMTV(BALB/cV), which encode V beta 2-specific superantigens. Furthermore, backcross female mice carrying Mtv-48 but not Mtv-51 were found to be able to produce milk-borne MMTV(CS), which can vigorously stimulate V beta 2-expressing T cells after local injection in vivo in an I-E dependent manner. On the other hand, mice carrying Mtv-51 but not Mtv-48 could not produce such an MMTV in milk. The nucleotide sequences of MMTV(CS) open reading frame were completely matched with those of Mtv-48. These results indicate that the provirus Mtv-48 but not Mtv-51 is capable of producing a milk borne virus of which the superantigen stimulates V beta 2-expressing T cells. PMID- 7474152 TI - Peptide vaccine against canine parvovirus: identification of two neutralization subsites in the N terminus of VP2 and optimization of the amino acid sequence. AB - The N-terminal domain of the major capsid protein VP2 of canine parvovirus was shown to be an excellent target for development of a synthetic peptide vaccine, but detailed information about number of epitopes, optimal length, sequence choice, and site of coupling to the carrier protein was lacking. Therefore, several overlapping peptides based on this N terminus were synthesized to establish conditions for optimal and reproducible induction of neutralizing antibodies in rabbits. The specificity and neutralizing ability of the antibody response for these peptides were determined. Within the N-terminal 23 residues of VP2, two subsites able to induce neutralizing antibodies and which overlapped by only two glycine residues at positions 10 and 11 could be discriminated. The shortest sequence sufficient for neutralization induction was nine residues. Peptides longer than 13 residues consistently induced neutralization, provided that their N termini were located between positions 1 and 11 of VP2. The orientation of the peptides at the carrier protein was also of importance, being more effective when coupled through the N terminus than through the C terminus to keyhole limpet hemocyanin. The results suggest that the presence of amino acid residues 2 to 21 (and probably 3 to 17) of VP2 in a single peptide is preferable for a synthetic peptide vaccine. PMID- 7474153 TI - A point mutation in the VP4 coding sequence of coxsackievirus B4 influences virulence. AB - In analyzing the molecular determinants of virulence of coxsackievirus B4, chimeric viruses were constructed from avirulent and virulent viruses. The vCB424 recombinant contained a single nucleotide substitution on an avirulent genetic background, resulting in replacement of Ser-16 of VP4 with Arg-16. Mice infected with vCB424 displayed an intermediate phenotype. PMID- 7474154 TI - Activation of human immunodeficiency virus type 1 in monocytoid cells by the protozoan parasite Leishmania donovani. AB - In this study, we demonstrated that the protozoan parasite Leishmania donovani and one of its major surface molecules, the lipophosphoglycan (LPG), can induce human immunodeficiency virus type 1 (HIV-1) expression in U1 and OM-10.1, two cell lines of monocytoid origin latently infected with HIV-1. Treatment of U1 cells with various concentrations of LPG (1, 5, and 10 microM) resulted in a dose dependent secretion of tumor necrosis factor alpha (TNF-alpha). Suppression of LPG-induced HIV-1 expression by polyclonal anti-TNF-alpha antibodies further confirmed the involvement of this cytokine. Results from these studies indicate that the protozoan parasite L. donovani can induce the secretion of TNF-alpha that will function in an autocrine or paracrine manner to upregulate HIV-1 expression. Our data suggest for the first time that this protozoan parasite can be viewed as a potential cofactor in the pathogenesis of AIDS. PMID- 7474155 TI - Gamma interferon is critical for resistance to Theiler's virus-induced demyelination. AB - Administration of neutralizing monoclonal antibody to gamma interferon increased Theiler's virus-induced demyelination and virus antigen persistence in the spinal cord in susceptible SJL/J mice and completely abrogated resistance such that all C57BL/10SNJ mice developed demyelination. These experiments support the hypothesis that gamma interferon is critically important for resistance to Theiler's virus-induced disease but is not required for myelin destruction. PMID- 7474156 TI - Initiator elements and regulated expression of the herpes simplex virus thymidine kinase gene. AB - Regulated expression of thymidine kinase mRNA in herpes simplex virus mutants harboring thymidine kinase promoters that lacked functional TATA boxes was largely unaffected by additional sequence alterations around the transcriptional start site. A strong initiator element increased the regulated expression of a TATA-containing promoter by 50% but did not affect that of the TATA-less promoter. Thus, initiator elements exert only small effects in this promoter context. PMID- 7474157 TI - Cell cycle dependence of foamy retrovirus infection. AB - In common with oncoviruses but unlike the lentivirus human immunodeficiency virus type 1, foamy (spuma) viruses require host cell proliferation for productive infection. We show that human immunodeficiency virus type 1 replicates in RD-CD4 cells regardless of the growth arrest condition of the cells, while murine leukemia virus is unable to infect growth-arrested RD-CD4 cells or cells progressing through a partial cell cycle that includes S phase but not mitosis. Human foamy virus, like murine leukemia virus, does not productively infect G1/S or G2 growth-arrested cells. Two other foamy viruses, simian foamy virus type 1, isolated from a macaque, and simian foamy virus type 6, isolated from a chimpanzee, also fail to establish productive infection in G1/S-arrested cells. PMID- 7474158 TI - Inhibition of murine retrovirus-induced neurodegeneration in the spinal cord by explant culture. AB - The neurovirulent chimeric mouse ecotropic retrovirus FrCasE causes a rapid neurodegenerative disease of the central nervous system (CNS) characterized by the appearance of spongiform lesions in motor areas 10 days after neonatal inoculation. To study the details of the pathogenic process, we examined the ability of an ex vivo spinal cord model to recapitulate disease. Organotypic spinal cord slice cultures were established from IRW mice 7 days after neonatal inoculation. This corresponds to a time when virus expression in the CNS is first detectable but spongiform changes have yet to evolve. Infectivity associated with these cultures peaked at 7 days in vitro and persisted at this level for 6 weeks. FrCasE infection of the spinal cord slices was primarily found associated with microglial cells. Infection of neurons, astrocytes, oligodendroglia, and endothelial cells was not observed; however, significant astrogliosis was found. Despite the presence of extensive microglial infection in close association with spinal motor neurons in organotypic cultures, no virus-specific spongiform degenerative changes were observed. These results suggest that removal of motor neurons from the developing CNS, despite maintaining the local cytoarchitectural relationships, prevents the virus from eliciting its pathological effects. Possible reasons for the interruption of lesion development are discussed. PMID- 7474159 TI - Elements of the polyomavirus replication origin required for homologous recombination mediated by large T antigen. AB - We introduced various elements of the polyomavirus origin of DNA replication into the genome of rat cells, and we analyzed their capacity to elicit rearrangements within the integrated sequences when exposed to large T antigen. The cis-acting sequences required for homologous recombination were those that make up a functional replication origin. PMID- 7474160 TI - The lytic cycle of Epstein-Barr virus in the nonproducer Raji line can be rescued by the expression of a 135-kilodalton protein encoded by the BALF2 open reading frame. AB - In Epstein-Barr virus (EBV)-carrying nonproducer Raji cells, the induction of the viral replicative cycle by chemical treatment is limited to only the early stage and viral DNA synthesis is totally inhibited. We previously showed the absence of two messenger RNAs that are encoded by the BamHI-A fragment of the EBV genome and that correspond to open reading frames BALF2 and BARF1 in chemically induced Raji cells. Since the BALF2 gene encodes a 135-kDa DNA-binding protein which was immunoprecipitated by antibody against ICP8 protein, a key protein in herpes simplex virus replication, we asked whether the lack of productive cycle in Raji cells is due to the absence of expression of the BALF2 gene. We transfected the Raji cell line with the BALF2 gene. After chemical induction, the BALF2 transfected cells expressed not only early antigens but also late antigens. In these cultures, the viral particles were detected by electron microscopy. The expression of late antigens was completely inhibited by arabinofuranosylthymine, which is a specific inhibitor of viral DNA replication. The BALF2 gene might play an essential role in the induction of the EBV-lytic cycle. PMID- 7474161 TI - Identification of a major T-cell epitope within VP3 amino acid residues 24 to 37 of Theiler's virus in demyelination-susceptible SJL/J mice. AB - Intracerebral inoculation of susceptible strains of mice with Theiler's murine encephalomyelitis virus (TMEV) results in a chronic, immunologically mediated demyelinating disease that shares many features with human multiple sclerosis. CD4+ T lymphocytes play a critical role in the pathogenesis of virus-induced demyelinating disease. We have identified a region within amino acid residues 24 to 37 of the VP3 capsid protein of TMEV (VP3(24-37)) that is recognized by T lymphocytes from the demyelination-susceptible SJL/J strain of mice. The T-cell response to VP3(24-37) represents a predominant Th-cell response against the virus from either TMEV-immunized or TMEV-infected SJL/J mice, and viral epitopes VP1(233-250), VP2(74-86), and VP3(24-37) account for most of the Th-cell response to TMEV. PMID- 7474162 TI - The packaging phenotype of the SE21Q1b provirus is related to high proviral expression and not trans-acting factors. AB - The avian packaging cell line SE21Q1b produces particles which encapsidate cellular RNAs. Such RNAs can be reverse transcribed by endogenous polymerase and integrated into the genomes of newly infected cells (M. Linial, Cell 49:93-102, 1987). Genomic RNA is not packaged because the packaging (psi) region of the provirus is deleted. The provirus also lacks the negative-strand primer binding site, which prevents efficient reverse transcription of randomly packaged genomic RNA. Previous work from our laboratory suggested that the trans-acting defect which allows packaging of cellular mRNA mapped to the provirus but did not map to the nucleocapsid region of the gag gene (D.J. Anderson, P. Lee, K. L. Levine, J. Sang, S. A. Shah, O. O. Yang, P. R. Shank, and M. L. Linial, J. Virol. 66:204 216, 1992). We have found, using proviral recombinants between SE21Q1b and wild type Rous sarcoma virus, that packaging of cellular RNAs does not map to the gag gene. Rather, the propensity of SE21Q1b particles to package cellular mRNA is a function of the high level of particle production in these cells and not of any specific viral structural proteins. PMID- 7474163 TI - Enhanced splicing of nonconsensus 3' splice sites late during adenovirus infection. AB - The adenovirus major late transcription unit is an example of an alternatively spliced gene, in which a common 5' splice site can be spliced to more than 15 different 3' splice sites. Here we show that the specificity in 3' splice site recognition changes during virus infection, such that 3' splice sites with long consensus-type polypyrimidine tracts are repressed while 3' splice sites with short atypical polypyrimidine tracts, which bind U2AF65K inefficiently, are enhanced in splicing in late virus-infected nuclear extracts. On the basis of these experiments, we discuss a mechanism that helps to explain how the complex pattern of major late mRNAs is produced late during virus infection. PMID- 7474164 TI - Detection of feline immunodeficiency virus in semen from seropositive domestic cats (Felis catus). AB - Electroejaculates from experimentally infected domestic cats were evaluated for the presence of feline immunodeficiency virus (FIV). Virus was isolated from cell free seminal plasma and seminal cells by cocultivation with a feline interleukin 2-dependent CD4+ T-cell line, in which productive infection was demonstrated by syncytium formation and FIV gag p26 antigen secretion. In addition, an 868-bp segment of the FIV gag provirus gene was identified in cocultured cells by PCR and Southern analysis. A 582-bp fragment of the FIV gag provirus genome was detected by nested PCR and Southern analysis in nonfractionated seminal cells and in sperm purified by a swim-up procedure. This is the first report describing the detection of replication-competent FIV in cell-free and cell-associated forms in domestic cat semen. PMID- 7474165 TI - Determination of adeno-associated virus Rep68 and Rep78 binding sites by random sequence oligonucleotide selection. AB - To further define the canonical binding site for the P5-promoted Rep proteins of the adeno-associated virus, a modified random oligonucleotide selection procedure was performed, using purified recombinant Rep protein. These results may explain the effects of Rep on cellular gene expression. PMID- 7474166 TI - Point mutations in the herpes simplex virus type 1 Vmw110 RING finger helix affect activation of gene expression, viral growth, and interaction with PML containing nuclear structures. AB - Herpes simplex virus type 1 immediate-early protein Vmw110 (also known as ICP0) has been implicated in the control of the balance between the lytic and latent states, but the precise mechanisms by which it exerts its effects are unknown. Vmw110 includes a characteristic zinc binding domain, termed the C3HC4 domain or RING finger, which is essential for its function. The solution structure of a related herpesvirus RING finger domain suggested that an amphipathic alpha helix might be an important functional component of the RING finger. In this paper, we show that the equivalent region of Vmw110 is important for virus growth in tissue culture and for the normal interaction of Vmw110 with nuclear structures which include the PML protein. PMID- 7474167 TI - Antiviral activity of alpha interferon in Sindbis virus-infected cells is restored by anti-E2 monoclonal antibody treatment. AB - Pretreatment of AT3 rat prostatic carcinoma cells expressing the inhibitor of apoptosis bcl-2 (AT3-bcl-2 cells) with alpha interferon (IFN-alpha) affected replication of a virulent strain of Sindbis virus (SV) but did not protect against virus-induced cell death. Treatment of cells with IFN-alpha late during infection affected ongoing SV replication very little. Previous studies have shown that cross-linking of the viral glycoprotein E2 with antibody delays the inhibition of K+ influx by improving the function of Na+K+ATPase and the Na(+) K(+)-2Cl-cotransport system in SV-infected cells (P. Despres, J. W. Griffin, and D. E. Griffin, J. Virol. 69:7006-7014, 1995). In these studies, we have shown that treatment of infected cells with anti-E2 monoclonal antibody also restored the ability of IFN-alpha to induce antiviral activity in infected cells late during infection. The very low rate of virus release in SV-infected cells treated simultaneously with anti-E2 monoclonal antibody and IFN-alpha was postulated to be linked to inhibition of virus maturation. Synergistic effects of antibody and IFN-alpha are likely to be important for control of SV replication in vivo. PMID- 7474169 TI - Mode of spread to and within the central nervous system after oral infection of neonatal mice with the DA strain of Theiler's murine encephalomyelitis virus. AB - Theiler's murine encephalomyelitis virus is a neurotropic enterovirus known to cause biphasic neural disease after intracerebral inoculation into adult mice. The present study characterizes a neonatal mouse model with a high disease incidence for the study of the acute phase of the pathogenesis of the DA strain of Theiler's murine encephalomyelitis virus after oral infection. The route of viral spread to and within the central nervous system (CNS) was determined by examining the kinetics of viral replication in various organs and by performing histopathological analysis. Viral antigen was detected widely in the neonatal CNS, mainly in the gray matter, and it was asymmetrical and multifocal in its distribution, with considerable variation in lesion distribution from animal to animal. Necrotizing lesions appeared to expand by direct extension from infected cells to their close neighbors, with a general disregard of neuroanatomical boundaries. The diencephalon showed particular susceptibility to viral infection. Other areas of the CNS, including the cerebellum and dentate gyrus of the hippocampus, were consistently spared. Neurons with axons extending peripherally to other organs or receiving direct input from the peripheral nervous system were not preferentially affected. The kinetics of viral replication in the liver, spleen, and CNS and the histopathological findings indicate that viral entry to the CNS is via a direct hematogenous route in orally infected neonatal mice and that the disease then progresses within the CNS mainly by direct extension from initial foci. PMID- 7474168 TI - Regulatory genes of simian immunodeficiency viruses from west African green monkeys (Cercopithecus aethiops sabaeus). AB - The high seroprevalence of simian immunodeficiency viruses (SIVs) in African green monkeys (AGMs) without immunological defects in their natural hosts has prompted consideration of SIV-infected AGMs as a model of apathogenic SIV infection. Study of the molecular mechanisms of SIVagm asymptomatic infection could thus provide clues for understanding the pathogenesis of human immunodeficiency viruses. Regulatory genes could be candidates for genetic control of SIVagm apathogenicity. We have characterized Vpr, Tat, Rev, and Nef genes of two SIVagm strains isolated from naturally infected sabaeus monkeys captured in Senegal. The results provide further evidence that SIVagm from West African green monkeys is the most divergent class of AGM viruses, with structural features in long terminal repeat sequences and Vpr and Tat genes that distinguish them from viruses isolated from other AGM species (vervet, grivet, and tantalus monkeys). PMID- 7474171 TI - The varicella-zoster virus (VZV) open reading frame 47 (ORF47) protein kinase is dispensable for viral replication and is not required for phosphorylation of ORF63 protein, the VZV homolog of herpes simplex virus ICP22. AB - To investigate the role of varicella-zoster virus (VZV) open reading frame 47 (ORF47) protein kinase during infection, a VZV mutant was generated in which two contiguous stop codons were introduced into ORF47, thus eliminating expression of the ORF47 kinase. ORF47 kinase was not essential for the growth of VZV in cultured cells, and the growth rate of the VZV mutant lacking ORF47 protein was indistinguishable from that of parental VZV. Nuclear extracts from cells infected with parental VZV contained several phosphorylated proteins which were not detected in extracts from cells infected with the ORF47 mutant. The herpes simplex virus type 1 (HSV-1) UL13 protein (the homolog of VZV ORF47 protein) is responsible for the posttranslational processing associated with phosphorylation of HSV-1 ICP22 (the homolog of VZV ORF63 protein). Immunoprecipitation of 32P labeled proteins from cells infected with parental virus and those infected with ORF47 mutant virus yielded similar amounts of the VZV phosphoproteins encoded by ORF4, ORF62, ORF63, and ORF68 (VZV gE), and the electrophoretic migration of these proteins was not affected by the lack of ORF47 kinase. Therefore, while the VZV ORF47 protein is capable of phosphorylating several cellular or viral proteins, it is not required for phosphorylation of the ORF63 protein in virus infected cells. PMID- 7474170 TI - Herpes simplex virus capsids assembled in insect cells infected with recombinant baculoviruses: structural authenticity and localization of VP26. AB - Recently, recombinant baculoviruses have been used to show that expression of six herpes simplex virus type 1 genes results in the formation of capsid-like particles. We have applied cryoelectron microscopy and three-dimensional image reconstruction to establish their structural authenticity to a resolution of approximately 2.7 nm. By comparing capsids assembled with and without the expression of gene UL35, we have confirmed the presence of six copies of its product, VP26 (12 kDa), around each hexon tip. However, VP26 is not present on pentons, indicating that the conformational differences between the hexon and penton states of the major capsid protein, VP5, extend to the VP26 binding site. PMID- 7474174 TI - AMA's Science Reporters Conference. PMID- 7474172 TI - Isolation of a highly cytopathic lentivirus from a nondomestic cat. AB - A feline immunodeficiency virus-like virus (FIV-Oma) isolated from a Pallas' cat (Otocolobus manul) is highly cytopathic in CrFK cells, in contrast to the chronic, noncytolytic infection established by an FIV isolate from a domestic cat (FIV-Fca). The virions have typical lentivirus morphology, density, and magnesium dependent reverse transcriptase activity. The major core protein is antigenically cross-reactive with that of FIV-Fca; however, FIV-Oma transcripts do not cross hybridize with FIV-Fca. A conserved region of the FIV-Oma pol gene has 76 to 80% nucleic acid identify with the corresponding pol regions of other feline lentiviruses and 64 to 69% identity with those of human, ovine, and equine lentiviruses. PMID- 7474173 TI - The bovine herpesvirus 1 maturational proteinase and scaffold proteins can substitute for the homologous herpes simplex virus type 1 proteins in the formation of hybrid type B capsids. AB - We determined the nucleotide sequence of a 3.5-kb region of the bovine herpesvirus 1 (BHV-1) genome which contained the complete BHV-1 homologs of the herpes simplex virus type 1 (HSV-1) UL26 and UL26.5 genes. In HSV-1, the UL26 and UL26.5 open reading frames encode scaffold proteins upon which viral capsids are assembled. The UL26-encoded protein is also a proteinase and specifically cleaves both itself and the UL26.5-encoded protein. The overall BHV-1-encoded amino acid sequence showed only 41% identity to the HSV-1 sequences and was most divergent in the regions defined to be involved in the scaffolding function. We substituted the proteins encoded by the BHV-1 homologs of the UL26 and UL26.5 open reading frames, expressed in baculovirus, for the corresponding HSV-1 proteins in an in vitro HSV-1 capsid assembly system. The proteins expressed from the BHV-1 UL26 and UL26.5 homologs facilitated the formation of hybrid type B capsids indistinguishable from those formed entirely with HSV-1-encoded proteins. PMID- 7474175 TI - Another health care idea: disease management. PMID- 7474176 TI - ACIP moves ahead with plans for use of IPV. PMID- 7474177 TI - From the Agency for Health Care Policy and Research. PMID- 7474178 TI - From the Centers for Disease Control and Prevention. Use of mammography services by women aged > or = 65 years enrolled in Medicare--United States, 1991-93. PMID- 7474179 TI - From the Centers for Disease Control and prevention. Hypertension among Mexican Americans--United States, 1982-84 and 1988-91. PMID- 7474180 TI - Reducing dietary intake of fat and cholesterol in children. PMID- 7474181 TI - Reducing dietary intake of fat and cholesterol in children. PMID- 7474182 TI - Reducing dietary intake of fat and cholesterol in children. PMID- 7474183 TI - Medicare funding of nursing education. PMID- 7474184 TI - Insulin resistance and non-insulin-dependent diabetes. PMID- 7474185 TI - A 72-year-old man with localized prostate cancer. PMID- 7474186 TI - Diagnosis of Lyme disease. PMID- 7474187 TI - Family violence and the insurance industry: education, not discrimination. PMID- 7474188 TI - The effect of gaps in health insurance on continuity of a regular source of care among preschool-aged children in the United States. AB - OBJECTIVE: To estimate the prevalence and length of gaps in health insurance coverage and their effect on having a regular source of care in a national sample of preschool-aged children. DESIGN: Follow-up survey of a nationally representative sample of 3-year-old children in the US population by phone or personal interview. PARTICIPANTS: A total of 8129 children whose mothers were interviewed for the 1991 longitudinal Follow-up to the National Maternal and infant Health Survey. MAIN OUTCOME MEASURES: Report of any gap in health insurance for the children, the length of the gap, and the number of different sites where the children were taken for medical care as a measure of continuity of a regular source of care. RESULTS: About one quarter of Us children were without health insurance for at least 1 month during their first 3 years of life. Over half of these children had a health insurance gap of more than 6 months. Less than half of US children had only one site of care during their first 3 years. Children with health insurance gaps of longer than 6 months were at increased risk of having more than one care site (odds ratio = 1.52; 95% confidence interval, 1.19 to 1.96). This risk further increased when an emergency treatment was discounted as a multiple site of care. CONCLUSIONS: Having a gap in health insurance coverage is an important determinant for not having a regular source of care for preschool-aged children. This finding is of concern, given the sizable percentage of children in the United States who lacked continuous health care coverage during a critical period of development. PMID- 7474189 TI - Outcomes of patients with hypertension and non-insulin dependent diabetes mellitus treated by different systems and specialties. Results from the medical outcomes study. AB - OBJECTIVE: To compare the outcomes of patients with hypertension and non-insulin dependent diabetes mellitus (NIDDM) who were cared for in three different systems of care and by generalist and subspecialist physicians. DESIGN: An observational study with follow-up at three periods: (1) a 2-year study of 532 patients with hypertension and 170 patients with NIDDM who had entrance and exit histories, physical examinations, and laboratory tests; (2) a 4-year follow-up of 1044 patients with hypertension and 317 patients with NIDDM based on patient-reported functional status; and (3) 7-year mortality for 1296 patients with hypertension and 424 patients with NIDDM. SETTING AND PARTICIPANTS: Patients sampled from health maintenance organizations, large multispecialty groups, and solo or single specialty group practices in Boston, Mass, Los Angeles, Calif, and Chicago, Ill. Patients were designated as belonging to one of three systems of care: fee for service; prepaid patients in solo or small single-specialty groups or in large multispecialty group practices, referred to as independent practice associations; and staff-model health maintenance organizations. The principal providers were family practitioners, general internists, cardiologists, or endocrinologists. MAIN OUTCOME MEASURES: Physiological, functional, and mortality. For hypertension, we measured blood pressure and stroke incidence. For NIDDM, we measured blood pressure, glycosylated hemoglobin level, visual function, vibration sense, ulcers and infections in the feet, and albumin excretion rate. Functional outcomes were assessed using the Medical Outcomes Study 36-Item Short Form Health Survey (SF-36). Mortality was assessed for the 7 years following the entrance examination. RESULTS: We found no evidence that any one system of care or physician specialty achieved consistently better 2-year or 4-year outcomes than others for patients with NIDDM or hypertension. Endocrinologists appeared to achieve better foot-ulcer and infection outcomes for patients with NIDDM, particularly when compared with family practitioners. However, no other specialist differences were found in any individual measures for either condition. Moreover, no adjusted mortality differences among systems or among physicians specialties were observed in the 7-year follow-up period. CONCLUSION: No meaningful differences were found in the mean health outcomes for patients with hypertension or NIDDM, whether they were treated by different care systems or by different physician specialists. Although prepaid medicine relies more heavily on generalist physicians than does fee for service, there is no evidence from these analyses that the quality of care of moderately ill patients with these two common disease was adversely affected. These findings must be viewed in light of the historically higher costs of fee-for-service medicine and of subspecialty physician practice. PMID- 7474190 TI - Incidence of prostate cancer diagnosis in the eras before and after serum prostate-specific antigen testing. AB - OBJECTIVE: To estimate the incidence of prostate cancer in Olmsted County, Minnesota, from 1983 through 1992 to describe the secular changes that have occurred since the introduction of serum prostate-specific antigen (PSA) testing to the community medical practice in 1987. DESIGN: Population-based, descriptive epidemiological study with ecological and individual level comparisons over time. STUDY SETTING: Olmsted County, Minnesota, where the Rochester Epidemiology Project provides passive surveillance of the population for health outcomes. SUBJECTS: All 511 biopsy-proven incident cases of adenocarcinoma of the prostate diagnosed from 1983 through 1992. The community inpatient and outpatient medical records of all incident cases were reviewed to evaluate the presenting characteristics of men at the time of diagnosis. RESULTS: The age-adjusted incidence of biopsy-proven prostate cancer increased from 64 per 100,000 person years in 1983 to 216 per 100,000 person-years in 1992. The increase occurred primarily between 1987 and 1988 and was predominately for organ-confined tumors. The age-specific incidence increased dramatically in this same period among men aged 50 years and older. Among men aged 70 years and older, however, prostate carcinoma incidence rates declined after 1990 following the initial increase. This decline among older men contrasted with community-based estimates of PSA utilization rates, which demonstrated consistent increases since 1987 to nearly 50% of the older population in 1992. CONCLUSION: These results support the premise that the recent increase in prostate cancer is due in part to the increased utilization of serum PSA testing. Further, the increased incidence appears to be a transient phenomenon due to the depletion of previously undiagnosed cases from the previous pool. Finally, these data suggest that, in terms of stage at diagnosis, early detection efforts may be effective in identifying more early stage (smaller) cancers. PMID- 7474192 TI - Subverting randomization in controlled trials. AB - Recent empirical evidence supports the importance of adequate randomization in controlled trials. Trials with inadequate allocation concealment have been associated with larger treatment effects compared with trials in which authors reported adequate allocation concealment. While that provides empirical evidence of bias being interjected into trials, trial investigators rarely document the sensitive details of subverting the intended purpose of randomization. This article relates anonymous accounts run the gamut from simple to intricate operations, from transillumination of envelopes to searching for code in the office files of the principal investigator. They indicate that deciphering is something more frequent than a rate occurrence. These accounts prompt some methodological recommendations to help prevent deciphering. Randomized controlled trials appear to annoy human nature--if properly conducted, indeed they should. PMID- 7474191 TI - Body weight and low-density lipoprotein cholesterol changes after consumption of a low-fat ad libitum diet. AB - OBJECTIVE: To assess the effects of a diet restricted in fat, saturated fat, and cholesterol, under weight-maintenance and ad libitum conditions on body weight and plasma lipid levels in hypercholesterolemic subjects. DESIGN: Dietary intervention study. SETTING AND PARTICIPANTS: Twenty-seven free-living, healthy middle-aged and elderly men (n = 13, age range, 41 to 81 years) and women (n = 14, age range, 52 to 79 years) with moderate hypercholesterolemia (low-density lipoprotein cholesterol [LDL-C] > or = 3.36 mmol/L [130 mg/dL]) participated in the study. INTERVENTION: Subjects underwent three dietary phases. First, subjects were provided with a diet similar to the average US diet (baseline diet; 35.4% total fat, 13.8% to 14.1% saturated fat, and 30 to 35 mg/1000 kJ [128 to 147 mg/1000 kcal] cholesterol). During the second dietary phase, subjects consumed a low-fat diet (15.1% total fat, 5.0% saturated fat, 17 mg/1000 kJ [73 mg/1000 kcal] cholesterol). During the baseline and low-fat diet phases, which lasted 5 to 6 weeks each, the energy intake was adjusted to keep body weight constant. During the third diet phase (low-fat ad libitum diet) subjects were given the same low-fat diet for 10 to 12 weeks, but could adjust their intake between 66% and 133% of the energy required to maintain body weight. MAIN OUTCOME MEASURES: Body weight and plasma lipid levels. RESULTS: Consumption of the low-fat diet under weight-maintenance conditions had significant lowering effects on plasma total cholesterol (TC), LDL-C, and high-density lipoprotein cholesterol (HDL-C) levels (mean change, -12.5%, -17.1%, and -22.8%, respectively). This diet significantly increased plasma triglyceride levels (+47.3%) and the TC/HDL-C ratio (+14.6%). In contrast, consumption of the low-fat ad libitum diet was accompanied by significant weight loss (3.63 kg), by a mean decrease in LDL-C (124.3%), and by mean triglyceride levels and TC/HDL-C ratio that were not significantly different from values obtained at baseline. CONCLUSIONS: Our results indicate that a low-fat ad libitum diet promotes weight loss and LDL-C lowering without adverse effects on triglycerides or the TC/HDL-C ratio in middle aged and elderly men and women with moderate hypercholesterolemia. PMID- 7474193 TI - Intrafamilial transmission of hepatitis-C virus among the population of an endemic area of Japan. AB - OBJECTIVES: To assess the role of intrafamilial transmission of hepatitis C virus (HCV) among general populations. DESIGN AND SETTING: Cross-sectional study in an HCV-endemic area of Japan. PARTICIPANTS: A total of 1122 residents (mean age, 41.7 years; range, 0 to 80 years), including 359 mother-child pairs and 234 pairs of spouses. MAIN OUTCOME MEASURES: Antibody to HCV (anti-HCV) was examined using second-generation anti-HCV testing by passive hemagglutination assay. Hepatitis C virus RNA was detected by polymerase chain reaction with primers deduced from the 5'-noncoding region and HCV genotypes by reaction with type-specific primers deduced from the HCV core gene. RESULTS: Prevalence of anti-HCV was 14.1% (158/1122), and HCV RNA was detected in 82.9% of those who tested positive for anti-HCV. Prevalence of anti-HCV increased with advancing age, but no anti-HCV was found among 312 persons younger than 20 years. Of the 53 children with anti HCV-positive mothers, three were positive for anti-HCV, all were older than 20 years, and all had a history of surgical treatment. Hepatitis C virus RNA was found in one mother-child pair but with different HCV genotypes. Both spouses were positive for anti-HCV in 17 of 234 pairs. Of the 11 pairs of spouses in whom the HCV genotypes of both were determined, five had different HCV genotypes. CONCLUSIONS: While HCV is highly endemic in this area, neither vertical nor horizontal transmission between spouses seems to play an important role in its spread. The incidence of intrafamilial transmission of HCV seems to be low. PMID- 7474194 TI - Rationale for treatment of patients with chronic heart failure with adrenergic blockade. AB - OBJECTIVE: Chronic heart failure is the leading cause of hospital admissions in patients older than 65 years. Heart failure due to systolic dysfunction is accompanied by activation of the sympathetic nervous system that contributes to progressive symptoms and an increased risk of death. While several clinical trials have suggested that antagonizing this sympathetic activation with beta blocking agents may provide clinical benefit, no clear consensus exists regarding use of beta-blockers for congestive heart failure. Therefore, we review the pathophysiology of the sympathetic nervous system as a basis for examining these clinical trials in order to understand the rationale for beta-blockade as a treatment for heart failure. DATA SOURCE: English language journal articles and reviews from a MEDLINE search and abstracts published at major cardiology meetings that related either to pathophysiology of the sympathetic nervous system or to therapy of patients with chronic heart failure with beta-blockers. STUDY SELECTION: Uncontrolled trials describing the initial use of this therapy and the subsequent randomized, placebo-controlled trials of beta-blockers were included. CONCLUSIONS: Sympathetic nervous system activation in patients with chronic heart failure is a major contributor to the severity of disease as well as its progression over time. Antagonism of its effects, via beta-blocker therapy, appears overall to improve both quality of life and survival. However, its place as a cornerstone in the therapy of this disease depends on the results of large scale, randomized, placebo-controlled trials. PMID- 7474195 TI - The effect on children of curtailing Medicaid spending. PMID- 7474196 TI - Uninsured children. An unintended consequence of health care system reform efforts. PMID- 7474197 TI - Physician specialty, systems of health care, and patient outcomes. PMID- 7474198 TI - Health system deregulation. Some aspects of health care system reform need not be held hostage. PMID- 7474199 TI - New techniques resculpt the cornea. PMID- 7474200 TI - Proposed flat tax on vaccines seen likely to pass. PMID- 7474202 TI - From the Centers for Disease Control and Prevention. Progress toward poliomyelitis eradication--South East Asia region, 1988-1994. PMID- 7474201 TI - From the Joint Commission on Accreditation of Healthcare Organizations. PMID- 7474203 TI - From the Centers for Disease Control and Prevention. State-specific changes in physical activity among persons aged > or = 65 years--United States, 1987-1992. PMID- 7474204 TI - From the Centers for Disease Control and Prevention. State-specific pregnancy and birth rates among teenagers--United States, 1991-1992. PMID- 7474205 TI - Patching for corneal abrasions. PMID- 7474206 TI - Respiration disorders in panic attacks. PMID- 7474207 TI - Respiration disorders in panic attacks. PMID- 7474208 TI - Endarterectomy for asymptomatic carotid artery stenosis. PMID- 7474209 TI - Endarterectomy for asymptomatic carotid artery stenosis. PMID- 7474210 TI - Endarterectomy for asymptomatic carotid artery stenosis. PMID- 7474211 TI - Endarterectomy for asymptomatic carotid artery stenosis. PMID- 7474212 TI - Domestic violence against women. PMID- 7474213 TI - Domestic violence against women. PMID- 7474214 TI - Domestic violence against women. PMID- 7474215 TI - Geriatric psychiatry and the limits of MeSH. PMID- 7474216 TI - Human foamy virus and familial Mediterranean fever in Japan. PMID- 7474217 TI - Health care actors: the patients' feet are tied. PMID- 7474218 TI - Cognitive effects after epidural vs general anesthesia. PMID- 7474219 TI - Health-related quality of life in primary care patients with mental disorders. Results from the PRIME-MD 1000 Study. AB - OBJECTIVE: To determine if different mental disorders commonly seen in primary care are uniquely associated with distinctive patterns of impairment in the components of health-related quality of life (HRQL) and how this compares with the impairment seen in common medical disorders. DESIGN: Survey. SETTING: Four primary care clinics. SUBJECTS: A total of 1000 adult patients (369 selected by convenience and 631 selected by site-specific methods to avoid sampling bias) assessed by 31 primary care physicians using PRIME-MD (Primary Care Evaluation of Mental Disorders) to make diagnoses of mood, anxiety, alcohol, somatoform, and eating disorders. MAIN OUTCOME MEASURES: The six scales of the Short-Form General Health Survey and self-reported disability days, adjusting for demographic variables as well as psychiatric and medical comorbidity. RESULTS: Mood, anxiety, somatoform, and eating disorders were associated with substantial impairment in HRQL. Impairment was also present in patients who only had subthreshold mental disorder diagnoses, such as minor depression and anxiety disorder not otherwise specified. Mental disorders, particularly mood disorders, accounted for considerably more of the impairment on all domains of HRQL than did common medical disorders. Finally, we found marked differences in the pattern of impairment among different groups of mental disorders just as others have reported unique patterns associated with different medical disorders. Whereas mood disorders had a pervasive effect on all domains of HRQL, anxiety, somatoform, and eating disorders affected only selected domains. CONCLUSIONS: Mental disorders commonly seen in primary care are not only associated with more impairment in HRQL than common medical disorders, but also have distinct patterns of impairment. Primary care directed at improving HRQL needs to focus on the recognition and treatment of common mental disorders. Outcomes studies of mental disorders in both primary care and psychiatric settings should include multidimensional measures of HRQL. PMID- 7474220 TI - In utero exposure to phenobarbital and intelligence deficits in adult men. AB - OBJECTIVE: To test whether exposure to phenobarbital in utero is associated with deficits in intelligence scores in adult men and whether the magnitude of the postnatal effect is mediated by exposure parameters and/or postnatal environmental factors. DESIGN: Two double-blind studies were conducted on independent samples of adult men prenatally exposed to phenobarbital and matched control samples using different measures of general intelligence. Based on data from control subjects, regression models were built relating intelligence scores to relevant pre-exposure matching variables and age at testing. Models generated predicted scores for each exposed subject. Group mean differences between the individually predicted and observed scores estimated exposure effects. SETTING: Copenhagen, Denmark. PARTICIPANTS: Exposed subjects were adult men born at the largest hospital in Copenhagen between 1959 and 1961 who were exposed to phenobarbital during gestation via maternal medical treatment and whose mothers had no history of a central nervous system disorder and no treatment during pregnancy with any other psychopharmacological drug. Study 1 included 33 men and study 2, 81 men. Controls were unexposed members of the same birth cohort matched on a wide spectrum of maternal variables recorded prenatally and perinatally. Controls for studies 1 and 2 included 52 and 101 men, respectively. MAIN OUTCOME MEASURES: In study 1: Wechsler Adult Intelligence Scale (Danish version); in study 2: Danish Military Draft Board Intelligence Test (Borge Priens Prove). RESULT: Men exposed prenatally to phenobarbital had significantly lower verbal intelligence scores (approximately 0.5 SD) than predicted. Lower socioeconomic status and being the offspring of an "unwanted" pregnancy increased the magnitude of the negative effects. Exposure that included the last trimester was the most detrimental. CONCLUSION: Phenobarbital exposure during early development can have long-term deleterious effects on cognitive performance. Detrimental environmental conditions can interact with prenatal biological insult to magnify negative outcomes. Physicians are urged to use increased caution in prescribing such medications during pregnancy. PMID- 7474221 TI - Total plasma homocysteine and cardiovascular risk profile. The Hordaland Homocysteine Study. AB - OBJECTIVE: To estimate the relations between established cardiovascular risk factors and total homocysteine (tHcy) in plasma. DESIGN: Health examination survey by the Norwegian Health Screening Service in 1992 and 1993. SETTING: General community, Hordaland County of Western Norway. PARTICIPANTS: A total of 7591 men and 8585 women, 40 to 67 years of age, with no history of hypertension, diabetes, coronary heart disease, or cerebrovascular disease were included. MAIN OUTCOME MEASURE: Plasma tHcy level. RESULTS: The level of plasma tHcy was higher in men than in women and increased with age. In subjects 40 to 42 years old, geometric means were 10.8 mumol/L for 5918 men and 9.1 mumol/L for 6348 women. At age 65 to 67 years, the corresponding tHcy values were 12.3 mumol/L (1386 men) and 11.0 mumol/L (1932 women). Plasma tHcy level increased markedly with the daily number of cigarettes smoked in all age groups. Its relation to smoking was particularly strong in women. The combined effect of age, sex, and smoking was striking. Heavy-smoking men aged 65 to 67 years had a mean tHcy level 4.8 mumol/L higher than never-smoking women aged 40 to 42 years. Plasma tHcy level also was positively related to total cholesterol level, blood pressure, and heart rate and inversely related to physical activity. The relations were not substantially changed by multivariate adjustment, including intake of vitamin supplements, fruits, and vegetables. CONCLUSIONS: Elevated plasma tHcy level was associated with major components of the cardiovascular risk profile, ie, male sex, old age, smoking, high blood pressure, elevated cholesterol level, and lack of exercise. These findings should influence future studies on the etiology and pathogenesis of cardiovascular disease. PMID- 7474222 TI - Perspectives on the physician workforce to the year 2020. AB - Physician supply and demand for the period extending to 2020 were assessed from three perspectives: physician utilization in group- and staff-model health maintenance organizations, physician distribution, and the future supply of nonphysician clinicians. The national norm for physician demand in 1993 was estimated to be 205 per 100,000 population. Demand is projected to increase 18% by 2020, because of both an expansion in beneficial services and a reduction in physician work effort. Supply initially will increase more rapidly, resulting in a surplus of 31,000 physicians (5% of patient care physicians) in the year 2000 and increasing to 62,000 physicians (8%) in 2010, after which the gap will narrow. Similar results were obtained when previous studies that had projected surpluses of 73,000 to 165,000 physicians (15% to 30%) in the year 2000 were reevaluated. However, physician distribution is not homogeneous, and the number of physicians per capita currently varies by more than twofold among states. Relative to the national norm, surpluses already exist in some states and shortages in others. In addition, the supply of nonphysician clinicians with independent practice authority is increasing. Their numbers are projected to double by 2010, equaling 60% of the number of patient care physicians. Measured as physician equivalents, their growth will equal the growth in physician supply. Thus, in terms of physicians alone, there is no evidence of a major impending national surplus. Local surpluses will be influenced principally by how physicians distribute themselves geographically. The major determinant of overall physician surpluses in the future will be the extent to which patients continue to seek physicians for services that also will be offered by an expanded workforce of nonphysician clinicians. Policy is needed that encompasses the universe of clinicians who will be providing care to patients in the next century. PMID- 7474223 TI - Potentially ineffective care. A new outcome to assess the limits of critical care. AB - OBJECTIVE: To examine the limits of the effectiveness of critical care through the study of patients for whom it was ineffective. DESIGN: We studied the relationship between resource use and long-term outcome (2-year follow-up) in 402 consecutively admitted critical care patients to develop a benchmark for ineffective applications of critical care. We defined an outcome called potentially ineffective care (PIC), developed and evaluated a model with an independent data set to predict PIC from a patient's response to treatment, and estimated the economic effects of limiting care after a prediction of PIC. SETTING: The combined medical and surgical intensive care unit at a 600-bed university teaching hospital. PATIENTS: Two groups of 402 consecutively admitted critical care patients, one from 1989, the other from 1991. MAIN OUTCOME MEASURES AND RESULTS: Based on observations from a two-dimensional plot of resource use vs benefit for 402 critical care patients, PIC was defined as resource use in the upper 25th percentile and survival for less than 100 days after discharge. Thirteen percent of the patients fell into the PIC category and used 32% of the resources. A product of the APACHE risk estimates on days 1 and 5 of at least 0.35 predicted 37% of PIC outcomes with a specificity of 98%. In a second data set, PIC outcome prediction had a sensitivity of 43% and a specificity of 94%, and a positive predictive value of 80%. For the hospital studied, reduction of intensity of treatment after a prediction of a PIC outcome would result in a reduction of hospital charges in the range of $1.8 million to $5 million per year. CONCLUSION: Patients in the PIC category consumed a large portion of the resources devoted to critical care at an academic teaching hospital. We suggest a change in focus from assessment of the quality of critical care and risk-adjusted mortality to an assessment of ineffective care based on outcome and resource use and a patient's response to treatment over time. PMID- 7474224 TI - Dehydration. Evaluation and management in older adults. Council on Scientific Affairs, American Medical Association. AB - OBJECTIVE: To review published literature regarding dehydration in older individuals and formulate a consensus on the evaluation and treatment of this unrecognized cause of hospitalizations, morbidity, and mortality. DATA SOURCES AND STUDY SELECTION: The literature concerning dehydration in the elderly population from MEDLINE was reviewed from 1976 through 1995. Search terms included dehydration, elderly, evaluation, hospitalization, and treatment. Particular emphasis was placed on articles describing original research leading to the development of new information on the evaluation and treatment of dehydration and review articles relating to the epidemiology, detection, treatment and health outcomes of this syndrome common in the geriatric population, including frail, institutionalized individuals. DATA EXTRACTION: Data contributing to a broad scientific understanding of dehydration were initially grouped according to topic areas of the physiology of normal aging, illness associated clinical reports of dehydration in the elderly population, and diagnostic and therapeutic interventions. The authors developed a consensus based on the weight of evidence presented and the authors' experience in the field. CONCLUSIONS: Early diagnosis is sometimes difficult because the classical physical signs of dehydration may be absent or misleading in an older patient. Many different etiologies place the elderly at particular risk. In patients identified as being at risk for possible dehydration, an interdisciplinary care plan with regard to prevention of clinically significant dehydration is critical if maximum benefit is to result. PMID- 7474225 TI - Mental disorders. Quality of life and inequality of insurance coverage. PMID- 7474226 TI - Estimating physician workforce requirements. The devil is in the assumptions. PMID- 7474227 TI - Traffic safety experts prescribe treatment, sanctions to improve driving. PMID- 7474228 TI - Surgeons offer 'new lease' after domestic abuse. PMID- 7474230 TI - AIDS and human rights. PMID- 7474229 TI - Not everyone wants IPV; opposition gears up. PMID- 7474231 TI - From the Food and Drug Administration. PMID- 7474232 TI - From the Centers for Disease Control and Prevention. Unintentional carbon monoxide poisonings in residential settings--Connecticut, November 1993-March 1994. PMID- 7474233 TI - Quality of outpatient care: diabetes. PMID- 7474234 TI - Quality of outpatient care: diabetes. PMID- 7474235 TI - Quality of outpatient care: diabetes. PMID- 7474236 TI - Quality of chart review for quality of care. PMID- 7474237 TI - Surfing the Internet for patient information: the personal clinical web page. PMID- 7474238 TI - Rocky Mountain spotted fever: legal duty to warn for noncontagious disease. PMID- 7474239 TI - Medicine--more than molecules and money. PMID- 7474241 TI - Medicine--more than molecules and money. PMID- 7474240 TI - Medicine--more than molecules and money. PMID- 7474242 TI - Costs of oral anticoagulation treatment after myocardial infarction. PMID- 7474244 TI - Race, treatment, and long-term survival from prostate cancer in an equal-access medical care delivery system. AB - OBJECTIVE: To evaluate long-term survival of black and white prostate cancer patients in an equal-access medical care system to help distinguish biological from medical and social explanations of mortality differences. DESIGN AND SETTING: Retrospective study of US Department of Defense tumor registry patients with prostate cancer. Ethnicity, age, diagnosis, staging, risk factors, treatment, and survival end points were extracted. PATIENTS: Prostate cancer patients (N = 1606; 7.5% black, 92.5% white) who were active-duty personnel, dependents, or retirees eligible for care in the military medical system. MAIN OUTCOME MEASURES: Racial differences in tumor stage and grade, risk factors, recurrence, and treatment wait time (time between initial diagnosis and initial treatment); influence of stage, grade, treatment, wait time, age, and race on survival. RESULTS: No differences were found in behavioral risk factors or tumor grade or size, but blacks entered active treatment (P < .001) and exhibited a higher relative risk of cancer (P = .01) in younger age groups, presented with higher stage (P < .001), and demonstrated increased progression in distant metastatic disease (P = .01). No significant differences were detected in overall wait time. When adjusted for stage, no difference was found in type of treatment. Overall, stage, grade, and age were found to affect survival (P = .04 to P < .001), but race did not. When analyzed by stage, blacks demonstrated a clear trend of longer survival for distant metastatic disease (P = .04 to P = .06). This trend was confirmed using Kaplan-Meier estimates (P = .04, likelihood ratio). CONCLUSIONS: This analysis suggests that in an equal-access medical care system there are no stage-specific differences in treatment between black and white prostate cancer patients. Survival among blacks is similar to that among whites and may surpass it for high-stage disease. PMID- 7474243 TI - A controlled trial to improve care for seriously ill hospitalized patients. The study to understand prognoses and preferences for outcomes and risks of treatments (SUPPORT). The SUPPORT Principal Investigators. AB - OBJECTIVES: To improve end-of-life decision making and reduce the frequency of a mechanically supported, painful, and prolonged process of dying. DESIGN: A 2-year prospective observational study (phase I) with 4301 patients followed by a 2-year controlled clinical trial (phase II) with 4804 patients and their physicians randomized by specialty group to the intervention group (n = 2652) or control group (n = 2152). SETTING: Five teaching hospitals in the United States. PATIENTS: A total of 9105 adults hospitalized with one or more of nine life threatening diagnoses; an overall 6-month mortality rate of 47%. INTERVENTION: Physicians in the intervention group received estimates of the likelihood of 6 month survival for every day up to 6 months, outcomes of cardiopulmonary resuscitation (CPR), and functional disability at 2 months. A specifically trained nurse had multiple contacts with the patient, family, physician, and hospital staff to elicit preferences, improve understanding of outcomes, encourage attention to pain control, and facilitate advance care planning and patient-physician communication. RESULTS: The phase I observation documented shortcomings in communication, frequency of aggressive treatment, and the characteristics of hospital death: only 47% of physicians knew when their patients preferred to avoid CPR: 46% of do-not-resuscitate (DNR) orders were written within 2 days of death; 38% of patients who died spent at least 10 days in an intensive care unit (ICU); and for 50% of conscious patients who died in the hospital, family members reported moderate to severe pain at least half the time. During the phase II intervention, patients experienced no improvement in patient-physician communication (eg, 37% of control patients and 40% of intervention patients discussed CPR preferences) or in the five targeted outcomes, ie, incidence or timing of written DNR orders (adjusted ratio, 1.02; 95% confidence interval [CI], 0.90 to 1.15), physicians' knowledge of their patients' preferences not to be resuscitated (adjusted ratio, 1.22; 95% CI, 0.99 to 1.49), number of days spent in an ICU, receiving mechanical ventilation, or comatose before death (adjusted ratio, 0.97; 95% CI, 0.87 to 1.07), or level of reported pain (adjusted ratio, 1.15; 95% CI, 1.00 to 1.33). The intervention also did not reduce use of hospital resources (adjusted ratio, 1.05; 95% CI, 0.99 to 1.12). CONCLUSIONS: The phase I observation of SUPPORT confirmed substantial shortcomings in care for seriously ill hospitalized adults. The phase II intervention failed to improve care or patient outcomes. Enhancing opportunities for more patient-physician communication, although advocated as the major method for improving patient outcomes, may be inadequate to change established practices. To improve the experience of seriously ill and dying patients, greater individual and societal commitment and more proactive and forceful measured may be needed. PMID- 7474245 TI - Defensive medicine and obstetrics. AB - OBJECTIVE: To test the hypothesis that physicians with greater malpractice claims exposure, either through personal experience or in their practice environment, will use more prenatal resources and have a higher cesarean delivery rate than physicians with lesser claims exposure. DESIGN: Retrospective cohort study using county malpractice defendant rate data from the Washington State Physicians Insurance and Exchange Association and prenatal care, delivery method, and self reported obstetric suit experience data from the Content of Obstetrical Care Study database. SETTING: Washington State obstetric practices. PARTICIPANTS: Stratified random samples of obstetrician-gynecologists and family physicians. MAIN OUTCOME MEASURES: The rates of obstetric ultrasound use, referral and consultation, prenatal care resource use, and cesarean delivery. RESULTS: After controlling for patient, physician, and sociodemographic characteristics, we found no difference in prenatal resource use or cesarean delivery rate for low risk patients between physicians with more and less exposure to malpractice claims. CONCLUSIONS: This study does not support an association between the malpractice experience or exposure of individual physicians and an increase in the use of prenatal resources or ceserean deliveries for the care of low-risk obstetric patients. PMID- 7474247 TI - Carbon monoxide poisoning among recreational boaters. AB - OBJECTIVE: To describe the case characteristics of a series of patients poisoned with carbon monoxide (CO) while boating for recreation. DESIGN: Cases of patients referred for treatment of CO poisoning with hyperbaric oxygen were reviewed. Those cases that occurred during recreational boating were selected for analysis. SETTING: A private, urban, tertiary care center studied from July 1984 to June 1994. PATIENTS: Thirty-nine patients ranging in age from 6 months to 69 years who were poisoned in 27 separate incidents. MAIN OUTCOME MEASURES: Characteristics of the poisoning incidents were assessed at initial patient presentation, immediately following treatment, and with follow-up telephone interviews. RESULTS: Of 512 patients treated for acute unintentional CO poisoning, 39 cases (8%) occurred in 27 incidents related to recreational boating activities. Individuals typically lost consciousness as a result of the poisoning. Most cases occurred aboard a boat that was older than 10 years, had an enclosable cabin, was longer than 22 feet, was powered by a gasoline engine, and was without a CO detector on board. CONCLUSIONS: Carbon monoxide poisoning is a serious hazard associated with recreational boating. The installation of CO detectors aboard boat types typically associated with this syndrome should be strongly encouraged. PMID- 7474246 TI - Pharmacogenetic explanation for excessive beta-blockade following timolol eye drops. Potential for oral-ophthalmic drug interaction. AB - OBJECTIVE: To determine whether the effects of topically administered timolol in an individual would be dependent on the presence or absence in that individual of the P-450 enzyme CYP2D6 and whether the effects of topically administered timolol would be increased and its metabolism decreased by the oral administration of quinidine, a known inhibitor of CYP2D6. DESIGN: Single-blind randomized crossover comparison of topical timolol, placebo, and the effects of inhibition of timolol metabolism by oral quinidine. SETTING: Clinical research center of an academic medical center. PARTICIPANTS: Eight male extensive metabolizers (EMs) and five male poor metabolizers (PMs) of debrisoquin. INTERVENTION: Two drops of 0.5% timolol or artificial tears were administered into each nostril in random order, and placebo or 50 mg of quinidine was administered orally to the EMs in random order, followed 30 minutes later by either the timolol or placebo drops. MAIN OUTCOME MEASUREMENT: Plasma timolol concentrations were measured by high-pressure liquid chromatography, while the extent of beta-blockade was determined by the suppression of exercise-induced rise in heart rate. RESULTS: The exercise heart rate was reduced following timolol eye drops compared with placebo in both EMs (P < .001) and PMs (P < .001) with significantly greater heart rate reduction (P = .01) and higher plasma timolol concentration in PMs compared with EMs (P = .03). Administration of quinidine with timolol eye drops to EMs resulted in a further significant reduction in heart rate (P = .02) and increase in plasma timolol concentration (P = .04). CONCLUSIONS: An individual's debrisoquin phenotype is an important determinant of beta-blockade following timolol eye drops, and metabolism of timolol is inhibited and beta-blockade increased by coadministration of oral quinidine. Clinicians should be aware of the potential for drug interactions that occur when orally administered drugs inhibit the metabolism of a topically administered drug. PMID- 7474248 TI - The effect of sugar on behavior or cognition in children. A meta-analysis. AB - OBJECTIVE: To examine the effects of sugar on the behavior or cognition of children by using meta-analytic techniques on reported studies. DATA SOURCES: Studies were identified through a literature search of the MEDLINE and PsychINFO databases and the authors' files using sugar, sucrose, and attention deficit disorder as the search terms. STUDY SELECTION: Studies were required to (1) intervene by having the subjects consume a known quantity of sugar, (2) use a placebo (artificial sweetener) condition (3) blind the subjects, parents, and research staff to the conditions; and (4) report statistics that could be used to compute the dependent measures effect sizes. DATA EXTRACTION: Variables included publication year, study setting, subject type and number, gender, age, sugar and placebo type and dose, prior dietary condition, measurement construct, means and SDs for the sugar and placebo conditions, and direction of effect. DATA SYNTHESIS: Sixteen reports met the inclusion criteria for a total of 23 within subject design studies. The weighted mean effect size and related statistics for each of the 14 measurement constructs revealed that although the range for these means was from -0.14 for direct observations and up to +0.30 for academic tests, the 95% confidence interval for all 14 mean effect sizes included 0. CONCLUSION: The meta-analytic synthesis of the studies to date found that sugar does not affect the behavior or cognitive performance of children. The strong belief of parents may be due to expectancy and common association. However, a small effect of sugar or effects on subsets of children cannot be ruled out. PMID- 7474249 TI - Appropriateness of antiepileptic drug level monitoring. AB - OBJECTIVES: To develop explicit, reliable appropriateness criteria for antiepileptic drug level monitoring and to assess the appropriateness of monitoring in one tertiary care institution. DESIGN: Appropriateness criteria derived from the literature and through expert opinion were used to evaluate a stratified random sample of antiepileptic drug level determinations obtained from chart review. SETTING: Tertiary care center performing more than 10,000 antiepileptic drug level determinations per year. PATIENTS: A total of 330 inpatients in whom antiepileptic drug levels were measured a total of 855 times. METHODS: Drug levels were assessed at least 200 times for each of four antiepileptic drugs (phenytoin, carbamazepine, phenobarbital, and valproic acid). MAIN OUTCOME MEASURES: The proportion of antiepileptic drug levels with an appropriate indication and, of those, the proportion sampled appropriately. RESULTS: Overall, 27% (95% confidence interval, 24% to 30%) of levels had an appropriate indication. Interrater agreement for appropriateness was substantial (kappa = 0.61). There was no significant difference in the appropriateness rate among the four drugs (range, 25% to 29%). Of the 624 antiepileptic drug level determinations considered inappropriate (73%), only four (0.6%) were more than 20% higher than the upper limit of normal, and none of the four patients had clinical signs of drug toxicity. A median of six levels (range, one through 69) was determined per patient, and the median interval between level determinations was 24 hours. Of the 27% of level determinations with an appropriate indication, 51% were sampled correctly, resulting in an overall appropriateness rate of 14%. CONCLUSIONS: Only 27% of antiepileptic drug level determinations had an appropriate indication, and half of these were not sampled correctly. Routine daily monitoring without pharmacological justification accounted for most of the inappropriate drug level determinations. Efforts to decrease inappropriate monitoring may result in substantial cost reductions without missing important clinical results. PMID- 7474250 TI - Statement on use of apolipoprotein E testing for Alzheimer disease. American College of Medical Genetics/American Society of Human Genetics Working Group on ApoE and Alzheimer disease. AB - OBJECTIVE: To evaluate the published data on the association between apolipoprotein E genotype (APOE) and Alzheimer disease (AD) and determine whether the data support the use of genetic testing for diagnosis or prediction of disease. This statement is intended for neurologists, psychiatrists, geneticists, primary care providers, diagnostic laboratories, and the public. PARTICIPANTS: The joint American College of Medical Genetics (ACMG) and American Society of Human Genetics (ASHG) Test and Technology Transfer Committee developed a 10 member ACMG/ASHG Working Group to assess available data on the association of AD with APOE alleles. To ensure inclusion of clinical specialists primarily involved with AD patients and families, the American Academy of Neurology (AAN) and the American Psychiatric Association (APA) appointed liaisons to the Working Group. EVIDENCE: Peer-reviewed journal publications obtained from an Index Medicus search or known to members of the Working Group were the source of data on which the statement is based. CONSENSUS PROCESS: Following discussions with all members of the Working Group, a draft statement was prepared by the chair and circulated among all members until a consensus was reached. The consensus draft was sequentially reviewed and endorsed by the appropriate scientific and executive committees of the ACMG, ASHG, AAN, APA, and the National Institutes of Health Department of Education Working Group on Ethical, Legal, and Social Implications of Human Genome Research. In some instances, suggestions from these committees were incorporated into the final statement. CONCLUSIONS: There is general consensus that APOE epsilon 4 is strongly associated with AD and that when present may represent an important risk factor for the disease. However, at the present time it is not recommended for use in routine clinical diagnosis nor should it be used for predictive testing. Studies to date indicate that the APOE genotype alone does not provide sufficient sensitivity or specificity to allow genotyping to be used as a diagnostic test. Because AD develops in the absence of APOE epsilon 4 and because many with APOE epsilon 4 seem to escape disease, genotyping is also not recommended for use as a predictive genetic test. The results of a collaborative study under way will clarify some of these issues. Whether APOE genotypes have other uses in the management of AD will become apparent over the next few years. PMID- 7474251 TI - Users' guides to the Medical Literature. VIII. How to use clinical practice guidelines. B. what are the recommendations and will they help you in caring for your patients? The Evidence-Based Medicine Working Group. PMID- 7474252 TI - A cautionary note on market reforms in health care. PMID- 7474253 TI - Improving care near the end of life. Why is it so hard? PMID- 7474254 TI - The public's view of the future of Medicare. PMID- 7474255 TI - A piece of my mind. Another 'taste of lemonade'. PMID- 7474256 TI - Questions on genetic testing services. PMID- 7474257 TI - Military physicians lend healing hands to Haiti. PMID- 7474258 TI - Journal's aversion to prepublication publicity silences researchers at human genetics meeting. PMID- 7474259 TI - From the Centers for Disease Control and Prevention. Outbreak of acute febrile illness and pulmonary hemorrhage--Nicaragua, 1995. PMID- 7474260 TI - From the Centers for Disease Control and Prevention. Outbreak of salmonellosis associated with beef jerky--New Mexico, 1995. PMID- 7474261 TI - From the Centers for Disease Control and Prevention. Hospitalization for epilepsy -United States, 1988-1992. PMID- 7474262 TI - Preoperative cardiac evaluation and perioperative monitoring for noncardiac vascular surgery. PMID- 7474263 TI - Preoperative cardiac evaluation and perioperative monitoring for noncardiac vascular surgery. PMID- 7474264 TI - Myocardial injury in critically ill patients. PMID- 7474265 TI - Myocardial injury in critically ill patients. PMID- 7474266 TI - Handyperson's hazard: crawl space sporotrichosis. PMID- 7474267 TI - Complication rates as a measure of quality of care. PMID- 7474268 TI - The Postmenopausal Estrogen/Progestin Interventions Trial. PMID- 7474269 TI - The Postmenopausal Estrogen/Progestin Interventions Trial. PMID- 7474270 TI - Antilymphoblastic globulin in renal transplant patients: no allergic reactions: a correction. PMID- 7474271 TI - Inadequate functional health literacy among patients at two public hospitals. AB - OBJECTIVE: To determine the ability of patients to complete successfully basic reading and numeracy tasks required to function adequately in the health care setting. DESIGN: Cross-sectional survey. SETTING: Two urban, public hospitals. PATIENTS: A total of 2659 predominantly indigent and minority patients, 1892 English-speaking and 767 Spanish-speaking, presenting for acute care. MAIN OUTCOME MEASURE: Functional health literacy as measured by the Test of Functional Health Literacy in Adults (TOFHLA), an instrument that measures ability to read and understand medical instructions and health care information presented in prose passages and passages containing numerical information (eg, prescription bottle labels and appointment slips). RESULTS: A high proportion of patients were unable to read and understand written basic medical instructions. Of 2659 patients, 1106 (41.6%) were unable to comprehend directions for taking medication on an empty stomach, 691 (26%) were unable to understand information regarding when a next appointment is scheduled, and 1582 (59.5%) could not understand a standard informed consent document. A total of 665 (35.1%) of 1892 English speaking patients and 473 (61.7%) of 767 Spanish-speaking patients had inadequate or marginal functional health literacy. The prevalence of inadequate or marginal functional health literacy among the elderly (age > or = 60 years) was 81.3% (187/230) for English-speaking patients and 82.6% (57/69) for Spanish-speaking patients, and was significantly higher (P < .001) than in younger patients. CONCLUSIONS: Many patients at our institutions cannot perform the basic reading tasks required to function in the health care environment. Inadequate health literacy may be an important barrier to patients' understanding of their diagnoses and treatments, and to receiving high-quality care. PMID- 7474272 TI - Vitamin D deficiency in homebound elderly persons. AB - OBJECTIVE: To assess the vitamin D status in homebound, community-dwelling elderly persons; sunlight-deprived elderly nursing home residents; and healthy, ambulatory elderly persons. DESIGN: A cohort analytic study. PARTICIPANTS: Of 244 subjects at least 65 years old, 116 subjects (85 women and 31 men) had been confined indoors for at least 6 months, either in private dwellings in the community (the Hopkins Elder Housecall Program) or in a teaching nursing home (The Johns Hopkins Geriatrics Center). The 128 control subjects, a healthy ambulatory group, came from the Baltimore Longitudinal Study on Aging. All subjects were free of diseases or medications that might interfere with their vitamin D status. MAIN OUTCOME MEASURES: Serum levels of 25-hydroxyvitamin D (25 OHD) and 1,25-dihydroxyvitamin D (1,25-[OH]2D) were measured in all subjects. In a subgroup of 80 subjects, serum levels of intact parathyroid hormone (PTH), ionized calcium, and osteocalcin and intake of vitamin D (through 3-day food records) were assessed. A randomly selected cohort of sunlight-deprived subjects also had serum levels of vitamin D binding protein measured. RESULTS: In sunlight deprived subjects overall, the mean 25-OHD level was 30 nmol/L (12 ng/mL) (range, < 10 to 77 nmol/L [< 4 to 31 ng/mL]) and the mean 1,25-(OH)2D level was 52 pmol/L (20 pg/mL) (range, 18 to 122 pmol/L [7 to 47 pg/mL]). In the sunlight-deprived subjects, 54% of community dwellers and 38% of nursing home residents had serum levels of 25-OHD below 25 nmol/L (10 ng/mL) (normal range, 25 to 137 nmol/L [10 to 55 ng/mL]). A significant inverse relationship existed between 25-OHD (ie, Log [25-OHD]) and PTH when they were analyzed together (r = -0.42; R2 = 0.18; P < .001) and for each cohort separately. All other parameters measured, except ionized calcium, differed significantly from the Baltimore Longitudinal Study Group means. The mean (SD) daily intakes of vitamin D (121 [132] IU) and calcium (583 [322] mg) were below the recommended dietary allowance only in the community dwelling homebound population. The mean vitamin D binding protein level in the sunlight-deprived subgroup was in the normal range. CONCLUSIONS: Despite a relatively high degree of vitamin supplementation in the United States, homebound elderly persons are likely to suffer from vitamin D deficiency. PMID- 7474273 TI - Pitfalls in nonrandomized outcomes studies. The case of incidental appendectomy with open cholecystectomy. AB - OBJECTIVES: To assess the short-term outcomes of incidental appendectomy through analysis of hospital administrative data and determine the consistency and plausibility of the observed results. DESIGN: Population-based historical cohort study. SETTING: All general hospitals in Ontario between 1981 and 1990. PATIENTS: Patients undergoing open primary cholecystectomy with (7846 exposed) and without (191,599 unexposed) incidental appendectomy. MAIN OUTCOME MEASURES: In-hospital fatality rates, complication rates, and lengths of hospital stay. RESULTS: Crude comparisons showed a striking and paradoxical reduction in mortality after cholecystectomy when incidental appendectomy was performed (odds ratio [OR], 0.37; 95% confidence interval [CI], 0.23 to 0.57; P < .001); mean length of stay was also lower by -0.46 day (P < .001). After adjustment for confounding differences, such as comorbidity and nonelective surgery, mortality and lengths of stay were similar for exposed and unexposed patients; but exposed patients showed a significant increase in nonfatal complications (OR, 1.53; 95% CI, 1.39 to 1.68; P < .001). Adverse effects from incidental appendectomy emerged consistently for all three outcomes only after restricting the analysis to subgroups of patients at low surgical risk. The increased mortality for exposed patients was largest among low-risk groups; for example, among those younger than 70 years undergoing elective surgery, the OR was 2.65 (95% CI, 1.25 to 5.64; P < .001). CONCLUSION: These findings suggest that incidental appendectomy is associated with a small but definite increase in adverse postoperative outcomes. However, plausible and consistent findings were only obtained after restricting the analysis to low-risk subgroups in which unmeasured differences in patients' baseline characteristics were less likely to confound adjusted outcome comparisons. This exercise highlights the potential pitfalls in nonrandomized outcomes comparisons using data sources with limited clinical detail, such as hospital discharge abstracts. PMID- 7474274 TI - Nonsexual assaults to the genitals in the youth population. AB - OBJECTIVE: To assess the frequency with which youth suffer nonsexual assaults to the genitals and their context and consequences. DESIGN: Telephone survey with follow-up interview 1 year later. SETTING: General population of the United States living in households with telephones. PARTICIPANTS: Random sample of 1042 boys and 958 girls aged 10 through 16 years. RESULTS: A nonsexual assault to the genitals was experienced by 9.2% of the boys and 1.0% of the girls in the year prior to the initial interview and 9.1% of the boys and 2.2% of the girls in the year prior to the follow-up interview. Among the boys, about a quarter of the assaults involved some injury, but only one in 50 needed medical attention. The most common assailants were same-aged peers. The assaults occurred in a variety of contexts including gang attacks, peer fighting, bullying, and some situations in which girls retaliated against the genitals of harassing boys. Boy victims of nonsexual genital assault had significantly higher levels of posttraumatic and depression symptomatology than boys without such assaults. CONCLUSIONS: Nonsexual genital violence needs additional clinical and research attention. Youth should be educated about its possible consequences. Clinicians should ask about nonsexual genital violence when taking a history, particularly with youth who have experienced other kinds of assaults. PMID- 7474275 TI - Folate levels and neural tube defects. Implications for prevention. AB - Using data from a recent case-control study, a woman's risk of having a child with a neural tube defect (NTD) was found to be associated with early pregnancy red cell folate levels in a continuous dose-response relationship. These findings were used to calculate the reduction in NTD cases that would be expected under two different strategies to raise folate levels. Targeting high-risk individuals has a small effect on the population prevalence but can substantially change an individual's risk. Targeting the population produces a small change in individual risk but has a large effect on the population prevalence. Supplementation of high risk women would be the most efficient method to implement the high-risk strategy, while food fortification would be preferable for the population approach. The current guidelines for the prevention of NTD are for an increased folic acid intake of 0.4 mg per day. This would result in a 48% reduction in NTDs, which may be near optimal. The two intervention strategies should be considered complementary in prevention of NTDs. PMID- 7474276 TI - Utility of follow-up tests for detecting recurrent disease in patients with malignant melanomas. AB - OBJECTIVE: To determine the effectiveness of follow-up tests for signaling recurrences in patients with intermediate- and high-risk malignant melanomas treated with curative intention. DESIGN: Retrospective analysis of prospectively collected data. SETTING: North Central Cancer Treatment Group. PATIENTS: A total of 261 patients with resected local (> or = 1.69 mm) and regional nodal malignant melanomas who were enrolled in a single prospective adjuvant trial were studied. All patients were scheduled to be followed up monthly for 2 months, then every 2 months for the first year, every 4 months the second year, every 6 months the next 3 years, and annually thereafter, with each visit consisting of a history, physical examination, complete blood cell count, blood chemistry panel, and a chest x-ray. RESULTS: Of the 145 evaluable patients who developed recurrent melanomas, 99 patients (68%) developed symptoms that signaled the diagnosis of recurrent disease. Physical examination of asymptomatic patients led to the diagnosis of recurrent disease in 37 patients (26%). The other nine patients (6%) with recurrent disease had abnormal chest x-rays. Laboratory results were never a sole indicator of recurrent disease. CONCLUSION: The majority of recurrences following resection of primary melanomas are discovered by history and/or physical examination despite the frequent use of other follow-up tests. The present data indicate that routine blood analyses and chest x-rays have limited value in the postoperative follow-up of patients with resected intermediate- and high-risk melanomas. PMID- 7474277 TI - Antibiotic treatment of adults with infective endocarditis due to streptococci, enterococci, staphylococci, and HACEK microorganisms. American Heart Association. AB - OBJECTIVE: To provide guidelines for the treatment of endocarditis in adults caused by the following microorganisms: viridans streptococci and other streptococci, enterococci, staphylococci, and fastidious gram-negative bacilli of the HACEK group. PARTICIPANTS: An ad hoc writing group appointed by the American Heart Association under the auspices of the Committee on Rheumatic Fever, Endocarditis, and Kawasaki Disease, Council on Cardiovascular Disease in the Young. EVIDENCE: Published studies of the treatment of patients with endocarditis and the collective clinical experience of this group of experts. CONSENSUS PROCESS: The recommendations were formulated during meetings of the working group and were prepared by a writing committee after the group had agreed on the specific therapeutic regimens. The consensus statement was subsequently reviewed by standing committees of the American Heart Association and by a group of experts not affiliated with the working group. CONCLUSIONS: Sufficient evidence has been published that recommendations regarding treatment of the most common microbiological causes of endocarditis (viridans streptococci, enterococci, Streptococcus bovis, staphylococci, and the HACEK organisms) are justified. There are insufficient published data to make a strong statement regarding the efficacy of specific therapeutic regimens for cases of endocarditis due to microorganisms that uncommonly cause endocarditis. As a useful aid to the practicing clinician, the writing group developed a consensus opinion regarding management of endocarditis caused by the most commonly encountered microorganisms and regarding those cases due to infrequent causes of endocarditis. PMID- 7474278 TI - Female genital mutilation. Council on Scientific Affairs, American Medical Association. AB - Female genital mutilation is the medically unnecessary modification of female genitalia. Female genital mutilation typically occurs at about 7 years of age, but mutilated women suffer severe medical complications throughout their adult lives. Female genital mutilation most frequently occurs in Africa, the Middle East, and Muslim parts of Indonesia and Malaysia, and it is generally part of a ceremonial induction into adult society. Recent political and economic problems in these regions, however, have increased the numbers of students and refugees to the United States. Consequently, US physicians are treating an increasing number of mutilated patients. The Council on Scientific Affairs recommends that US physicians join the World Health Organization, the World Medical Association, and other major health care organizations in opposing all forms of medically unnecessary surgical modification of the female genitalia. PMID- 7474280 TI - Patients who can't read. Implications for the health care system. PMID- 7474279 TI - Urgent need to increase folic acid consumption. PMID- 7474281 TI - Academic physicians' participation in organized medicine. PMID- 7474282 TI - Medical students at the frontiers of research. PMID- 7474283 TI - The legacy of medical students in medical research. PMID- 7474284 TI - MD-PhD students in the 1990s: finding a niche for the dual degree. AB - Given the success of MD-PhD students in the past and the uncertain future of science and clinical medicine, this article has attempted to review the progress of MD-PhD programs and examine the experiences of some of today's MD-PhD students. Most candidates responding to the questionnaire remain committed to research and its applications. Their future plans are in line with the paths their predecessors have chosen, namely, careers devoted to academic medicine. For many of these dual-degree candidates, however, academia encompasses basic research and patient care. There seems to be little doubt that MD-PhD graduates will continue to seed the faculties of leading US medical schools and research institutes. However, some dual-degree recipients will opt for strictly clinical careers, perhaps leaving the laboratory altogether. Indeed, according to the author's survey, 14% of respondents intend to concentrate primarily on patient care. A few of these students seem disenchanted with the benefits conferred by a dual degree. "I don't see the advantages of a combined program that I saw when I was applying," one student explained. "The PhD degree confers no benefit as far as I can tell in how one practices medicine or in how well someone will conduct basic research." Detractors of combined-degree programs have questioned how anyone can succeed in science or medicine if not exclusively dedicated to one program. That achieving this balance was the foremost concern of the students surveyed suggests that doubt lingers for them as well. "I have often wished for more MD-PhD role models," wrote another student. "What have other people done with their combined degree? My career objectives are still vague because I am still not sure of what niche I will fill as an MD-PhD. Perhaps we can fill any niche." Perhaps that was the idea all along. PMID- 7474285 TI - Medicine in the humanities: recovering a tradition. PMID- 7474286 TI - Counselling: the big gap in disease management in Pakistan. PMID- 7474288 TI - Age and intraocular pressure: how are they correlated? AB - BACKGROUND: The relationship between intraocular pressure and age is not fully understood. Intraocular pressure may increase or decrease with age. PURPOSE: Study was planned to determine the effect of aging on intraocular pressure of an apparently healthy population of Karachi. SUBJECTS AND METHODS: Depending upon age, 8036 subjects were divided into seven groups. All were examined according to standard protocols. Measurements were taken by Goldmann applanation tonometer, after a complete rest for at least 15 minutes and always first in the right eye. RESULTS: Mean intraocular pressure increases progressively with age until 60 years. When one age group is compared with its preceding, the increase is non significant until 40 years but highly significant (P < 0.001) in the age groups of 41-50 and 51-60 years. The data shows non-significant decrease in the age group of 61-70 but an increase in the age group of above 70 years. CONCLUSION: This study supports the conclusion that as age increases, intraocular pressure also increases, with an average of 0.28 mmHg per decade. Knowledge of the normal range of intraocular pressure in various age groups will help glaucoma screeners. PMID- 7474287 TI - Ciprofloxacin in multi-resistant infections in childhood: an audit. AB - Ciprofloxacin is a new orally administrable fluoroquinolones, with considerable efficacy against multiresistant organisms. Its use in the paediatric age group however, is controversial because of the risk of potential articular toxicity. We retrospectively reviewed ciprofloxacin usage over a 32 week periods (June, 1991 September, 1993) in paediatric inpatients at The Aga Khan University Hospital. Ciprofloxacin was used in 21 cases, singly in 11 (52%) and in combination with other antibiotics in a further 10 (48%). The response to therapy was adjudged as 'good' or 'fair' in 13 (62%) cases. Ciprofloxacin was the only sensitive antibiotic in 4 (19%) and resistance to it was detected in another 4 (19%) cases. Despite all efforts, adequate follow-up could only be achieved in a third of the patients. Although no toxic or side effects were detected, in view of poor follow up and emergence of ciprofloxacin resistant strains, our experience highlights the need to regulate ciprofloxacin use in the paediatric age group. PMID- 7474289 TI - Effect of some psychoactive drugs on stress induced alteration in plasma corticosterone level. AB - Psychoactive drugs such as chlorpromazine, fluphenazine, haloperidol, propranolol and diazepam were evaluated for their ability to block stress induced changes in Wistar albino rats. The stress induced changes were monitored as the difference in plasma corticosterone (PCS) levels, before and after the administration of minimum effective doses of psychoactive drugs. Significant results were obtained with diazepam at the dose of 5-10 mg/kg and to a lesser extent with propranolol 20 mg/kg. Other drugs, at their minimum effective doses showed no significant change in plasma corticosterone levels. PMID- 7474290 TI - A new breakthrough in treatment of visceral leishmaniasis in children. AB - Fifty cases of visceral leishmaniasis were admitted in Children's Hospital, Islamabad. Common clinical features were fever (100%), splenomegaly (100%), hepatomegaly (100%), anaemia (96%), abdominal distension (40%), bronchopneumonia (26%) and bleeding diathesis (22%). Hb was below 7.0 G/dl in 80%, white cell count below 4 x 10(9)/cmm in 88% and platelet count below 100 x 10(9)/c4mm in 86%. All the patients showed leishmania donovani bodies in the marrow smears. Fourteen patients were treated with aminosidine (15 mg/kg), intramuscularly daily for 4 weeks. All responded dramatically and none of them went into relapse in a year's follow-up. No side-effects were observed. Aminosidine can therefore, be recommended as a treatment of choice for visceral leishmaniasis in children. PMID- 7474291 TI - Relative viscosity of plasma--evaluation of a simple technique. PMID- 7474292 TI - Effect of single dose of secnidazole in treatment of intestinal amoebiasis. PMID- 7474293 TI - Salicylate poisoning. AB - Salicylate poisoning continues to be an important cause of drug related mortality. The zero order kinetics of salicylates at high doses is responsible in part for cases of iatrogenic poisoning. Serum levels tend to correlate with severity of poisoning in acute overdose cases only. Clinical manifestations include involvement of nervous system, hepatic and pulmonary systems along with metabolic disturbances. Measures at enhancing elimination and reducing absorption, while providing supportive care form the basis of management. PMID- 7474294 TI - Sarcoidosis with skin manifestations--two case reports and review of literature. PMID- 7474295 TI - Hodgkin's disease presenting as nephrotic syndrome: a case report. AB - Nephrotic syndrome is a rare presentation of Hodgkin's disease. Majority of these cases have minimal change disease and some cases present as membranous glomerulopathy. This report presents the case of a child with nephrotic syndrome associated with Hodgkin's disease. PMID- 7474296 TI - Barium stone impaction in parkinsonism. AB - Autonomic symptoms such as orthostatic hypotension, abnormal sweating and constipation occur frequently in Parkinson's disease. In our case, barium meal used for upper gastrointestinal study caused barium stone formation and a paralytic-ileus-like syndrome. Therefore, attention should be paid while using barium meal for diagnostic purpose in Parkinsonism. PMID- 7474297 TI - Distal vasoconstriction following coronary angioplasty--comparison of emergent and elective PTCA. AB - To examine the occurrence of distal vasoconstriction following primary percutaneous transluminal coronary angioplasty (PTCA) for acute myocardial infarction, quantitative coronary angiography was performed during and after acute myocardial infarction. An assessment of coronary artery vasoconstriction was made in patients undergoing primary and elective PTCA. The primary PTCA group was subdivided into two groups based on initial antegrade flow characteristics: group 1 (TIMI grade flow 0/1) and group 2 (TIMI grade flow 2/3). All balloon angioplasties were performed during continuous administration of nitrates. There was no significant difference with relation to lesion diameter, lesion stenosis, or diameter of the left circumflex coronary artery between the acute and chronic phase in both groups. The distal diameter, however, was smaller during the acute phase than in the chronic phase in both groups (1.7 +/- 0.5 vs 2.0 +/- 0.4 mm, p < 0.001; 2.0 +/- 0.4 vs 2.2 +/- 0.3 mm, p < 0.05, respectively). The percent vasoconstrictive response of the distal segment was greater in the primary PTCA group than in the elective PTCA group (11.2 +/- 18.1 vs -1.9 +/- 7.1%, p < 0.0001, respectively). In conclusion, nitrate-resistant coronary artery vasoconstriction following primary PTCA occurs distal to the site of balloon dilation. PMID- 7474298 TI - CETP and LCAT activities are unrelated to smoking and moderate alcohol consumption in healthy normolipidemic men. AB - To distinguish between the effects of smoking and drinking on lipid metabolism, we conducted a cross-sectional study using 52 healthy normolipidemic subjects who either smoked or drank, but not both. The subjects were divided into three groups: Group S, smokers who did not drink (n = 20); Group D, drinkers who did not smoke (n = 12); and Group C, controls (n = 20), who neither drank nor smoked. High density lipoprotein cholesterol (HDL-C) levels and plasma cholesteryl ester transfer protein (CETP) and lecithin: cholesterol acyltransferase (LCAT) activities were measured in all of the subjects, and the values obtained in Group S and Group D were compared to those of controls. Group S had lower (p < 0.01) HDL2-C and HDL3-C levels, and Group D had higher (p < 0.05) HDL2-C and HDL3-C levels than the controls, but there were no significant differences between groups with respect to CETP and LCAT activities. Thus, in healthy normolipidemic men, both smoking and drinking affect HDL-C levels as expected, but do not affect plasma CETP or LCAT activity levels. PMID- 7474299 TI - Determinants of exercise capacity in the hypertensive elderly--a study based on the assessment of cardiac reserve by dipyridamole radionuclide angiography. AB - To evaluate the determinants of exercise capacity in the hypertensive elderly with an assessment of cardiac reserve by dipyridamole radionuclide angiography, 31 elderly subjects (aged 62 to 78 years, 15 hypertensive and 16 normotensive) underwent symptom-limited maximum treadmill exercise with measurement of maximal oxygen consumption along with radionuclide angiography in list mode at rest and after dipyridamole infusion. In the normotensive elderly, maximal oxygen consumption correlated with relative filling volume during the rapid filling period at rest (r = 0.58; p < 0.05), and correlated inversely with the percent change in the peak ejection rate with dipyridamole (r = -0.63; p < 0.01). In contrast, maximal oxygen consumption correlated with atrial contribution at rest (r = 0.69; p < 0.005), and correlated inversely with the percent change in atrial contribution with dipyridamole (r = -0.87; p < 0.0001) in the hypertensive elderly. These results indicate that the mechanisms for maintaining exercise capacity are different in elderly subjects with or without hypertension. PMID- 7474300 TI - A novel modified tissue-type plasminogen activator (t-PA), E6010, reduces reperfusion arrhythmias induced after coronary thrombolysis--comparison of native t-PA and urokinase. AB - The aim of this study was to compare the effects of a novel modified tissue-type plasminogen activator (t-PA), E6010, to those of native t-PA and urokinase on reperfusion arrhythmias (premature ventricular complexes: PVC) and the mortality rate after coronary thrombolysis. The frequency of PVC and the mortality rate were evaluated in anesthetized dogs which had 1-, 3- or 6-h-old thrombi induced by a copper coil in the coronary artery. Thrombolytic treatment with a bolus intravenous (iv) injection of E6010 was compared with the continuous iv administration of native t-PA or urokinase. The frequency of PVC was significantly lower in the E6010 group than in the native t-PA and urokinase groups (P < 0.05). The mortality rate in the E6010 group (0.0%) tended to be lower than those in the native t-PA group (10.7%) and the urokinase group (7.1%). These results indicate that the bolus iv injection of E6010 reduced both PVC and the mortality rate, compared with the continuous iv administration of native t-PA or urokinase for coronary thrombolysis; Therefore, E6010 may have beneficial effects in prehospital thrombolysis. PMID- 7474301 TI - Effects of suprachiasmatic lesions on circadian rhythms of blood pressure, heart rate and locomotor activity in the rat. AB - To determine whether the circadian rhythms in blood pressure (BP), heart rate (HR) and locomotor activity are controlled by an internal biological clock located in the suprachiasmatic nucleus (SCN), we continuously measured these parameters in SCN-lesioned rats using a newly developed implantable radiotelemetry device and a computerized data collecting system. Although SCN lesioned rats showed a weak but significant 24-h periodicity in BP and HR under light-dark (LD) cycles, BP, HR and locomotor activity became completely aperiodic under constant dark (DD) conditions. The amount of locomotor activity was significantly reduced in SCN-lesioned rats compared to that in intact rats. BP tended to be higher in SCN-lesioned rats, but the differences were significant only in the comparison of systolic blood pressure (SBP) under LD and DD (p < 0.05) and of mean blood pressure (MBP) under LD (p < 0.05). HR in SCN-lesioned rats was significantly lower under LD (p < 0.05), but not under DD. The standard deviation and the variation coefficient of MBP, as indices of short-term variability of this parameter, were significantly larger in SCN-lesioned rats than in intact rats, while those of HR and locomotor activity did not differ significantly between SCN-lesioned and intact rats. These results indicate that the SCN is important not only for generating circadian rhythms of BP, HR and locomotor activity, but also for buffering the short-term variability of BP in rats. PMID- 7474302 TI - Two adult cases of unilateral absence of the right pulmonary artery with markedly different clinical presentations. AB - Two adult cases of unilateral absence of the right pulmonary artery with markedly different clinical presentations are reported. One patient was a 21-year-old female without any known history of a coexisting congenital anomaly. An abnormal chest roentgenogram (small right hemithorax, deviation of the mediastinum toward the right side and a dilated left pulmonary artery) was noted and prompted further evaluation. No pulmonary hypertension was noted and the patient remained asymptomatic. The other patient was a 42-year-old male who had unilateral absence of the right pulmonary artery and a peripheral stenosis of the left pulmonary artery. The clinical course of this patient had been complicated by impaired exercise tolerance and occasional hemoptysis since adolescence. At the age of 29 years, a cardiac catheterization revealed pulmonary hypertension, but no left-to right shunt. Progressive respiratory failure resulted in a premature death at the age of 42 years. The prognosis of patients with unilateral absence of the pulmonary artery largely depends on the coexisting cardiac anomaly (left-to-right shunt) and pulmonary hypertension. A combination of unilateral absence of the pulmonary artery and contralateral peripheral pulmonary arterial stenosis is very rare, but is an important cause of pulmonary hypertension and gives a worse prognosis for this entity. PMID- 7474303 TI - A case of right atrial myxoma--effect of large myxoma in the right atrium studied by M-mode and Doppler echocardiography. AB - Doppler profiles are rarely used to assess cardiac function that has been partially impaired by a sizeable myxoma in the right atrium or to evaluate the improvement caused by extirpation of the tumor. In a 54-year-old man with a large right atrial myxoma (6.5 x 5.5 x 4.0 cm) along with first-degree atrioventricular (AV) block, M-mode and pulsed Doppler echocardiography were used to evaluate the left ventricular systolic and diastolic function before and 1 month after surgical removal of the myxoma. End-diastolic left ventricular (LV) and left atrial diameters increased postsurgically from 47 to 51 mm and from 38 to 41 mm, respectively, while end-systolic LV remained unchanged. In the LV inflow pattern, peak early filling velocity (E) increased substantially (preoperative 31, postoperative 58 cm/sec), with no change in peak late filling velocity (A) (53 cm/sec), which gave a favorable E/A ratio (from 0.58 to 1.09). First-degree AV block resolved after tumor resection (PR interval: 0.23 vs 0.20 sec). Improved LV diastolic function associated with natural recovery from the myxoma was ascribed to the restoration of preload and recovery of systolic function. The results of this study show that removal of a large myxoma in the right atrium is important not only for preventing possible obstruction of the tricuspid orifice, eliminating pulmonary emboli, and maintaining systolic function, but also for restoring LV diastolic function. PMID- 7474304 TI - [The analgesic effect of iontophoresis with magnesium sulfate]. AB - The analgesic effect of iontophoresis with magnesium sulfate was studied in fourteen healthy adult volunteers. Magnesium sulfate 0.5 mg.5 ml-1 was locally administered by the iontophoresis with 1.0 mA electric current for 10 min. The pain recognition time was measured by the pain thermometer before and after 5, 15, 30, 60 and 120 min of iontophoresis. The pain recognition time was significantly extended and lasted for 60 min. These results support the view that the iontophoresis with magnesium sulfate has analgesic effect and produces a good pain relief clinically. PMID- 7474305 TI - [The effects of sevoflurane on the coronary circulation in the instrumented dog with or without coronary stenosis]. AB - The effects of sevoflurane on the coronary circulation were investigated in the instrumented dogs with or without coronary stenosis. A-V block was produced by injection of formalin into A-V node to control heart rate at 120 beats.min-1 by pacing and mean systemic blood pressure was held constant at 65 mmHg with the pressurized blood reservoir. Left anterior descending coronary artery was cannulated with stainless steel tube supplied from the femoral artery. Total coronary flow was measured with an electromagnetic flow probe and the regional myocardial blood flow was calculated using color microsphere technique. Coronary resistance was reduced with the increasing end-tidal concentrations of sevoflurane up to 1.2 MAC. An adjustable screw clamp was used to reduce the coronary flow down to 50% of control value. Inner/outer ratio of regional myocardial blood flow decreased with stenosis, but was unaffected with 1.2 MAC of sevoflurane. We concluded that 1.2 MAC of sevoflurane was not strong enough to dilate the coronary artery to produce the transmural steal phenomenon in the dog with coronary stenosis. Further investigations are needed to clarify how hemodynamic factors influence the transmural steal phenomenon in myocardium. PMID- 7474306 TI - [Role of antigen-antibody reaction in pulmonary hypertensive reaction after antigen challenge]. AB - Pulmonary arterial pressure, airway pressure, and lung weight increase were studied after antigen (human O-N type erythrocytes) challenge in isolated perfused sensitized rabbit lungs. To investigate whether these hemodynamic changes are mainly caused by antigen-antibody reaction or by the other mechanisms, we measured changes of pulmonary arterial and airway pressures, and lung weight gain after antigen challenge in perfused nonsensitized rabbit lungs. Thirteen nonsensitized rabbits were divided into 2 groups; in N group (n = 5), antigen was given into Krebs Hanseleit perfusate; in Ab group (n = 8), antibody was given into reservoir 30 min prior to antigen challenge. Pulmonary arterial pressure in Ab group was higher than in N group after antigen challenge. Maximal increase in pulmonary arterial pressure after antigen challenge depended on agglutinin titer of antibody in perfusate (delta Ppa = 0.068 x [titer]-0.146, r2 = 0.929). However, there were no significant differences between the two groups in changes of airway pressure and lung weight gain after antigen challenge. Although agglutinin titer of perfusate in Ab group was higher than our previous study, pulmonary hypertensive reaction to antigen in Ab group was significantly lower. It is concluded that the other mechanism besides antigen-antibody reaction itself can be responsible for hemodynamic changes after antigen challenge. PMID- 7474307 TI - [The effects of systemic bupivacaine on baroreflex sensitivity in dogs]. AB - To evaluate the effects of systemic bupivacaine on the baroreflex control of heart rate, we investigated the baroreflex sensitivity assessed with a pressor and a depressor test using phenylephrine and nitroglycerin in pentobarbital anesthetized dogs (n = 12). Intravenous injection of bupivacaine (mean plasma concentration of 2.4 +/- 0.9 micrograms.ml-1) caused a significant suppression of the baroreflex sensitivity, defined by the slopes of regression line (in msec of RR interval change per mmHg increase or decrease in systolic blood pressure). The sensitivity obtained with the pressor and the depressor tests decreased from 6.0 +/- 2.3 to 3.5 +/- 1.7, from 2.4 +/- 1.3 to 1.3 +/- 0.8 msec.mmHg-1, respectively (P < 0.01). Suppression of the baroreflex sensitivity during epidural anesthesia with bupivacaine could be due not only to cardiac sympathectomy but also to a direct effect of bupivacaine on the reflex arch including the receptors, the afferent nerve pathways, the CNS, the efferent pathway, and the effector organs. Therefore, the hemodynamic responses to reduction of blood pressure are likely to be inhibited by epidural bupivacaine. PMID- 7474308 TI - [Intraoperative changes in blood lactate level and body temperature in younger and elderly adults]. AB - Sequential changes in arterial blood lactate concentration and central-peripheral temperature gradient during anesthesia were measured in 10 younger patients (46 63 yr) and 10 elderly patients (66-92 yr) undergoing abdominal surgery. A central temperature probe was attached at the rectum and peripheral probe on the palm of the hand. Anesthesia was induced with thiopental and succinylcholine and maintained with isoflurane and nitrous oxide in oxygen. There was a marked increase in lactate values intraoperatively in the elderly as compared to the younger patients. Difference between central and peripheral temperature increased significantly in the elderly patients, while there was no change in the temperature gradient in the younger patients. Central-peripheral temperature gradient is an indicator of peripheral circulation, and the results imply the presence of anaerobic oxidation in elderly patients during operation. In elderly patients, the functional reserve of organ system and the tolerance to surgical stress are considered to be reduced. Intraoperative elevation of blood lactate levels might be caused either by impairment of peripheral circulation or by metabolic deficit due to age-related decline in hepatic function. PMID- 7474309 TI - [Change of the cognitive function after open heart surgery]. AB - The changes of cognitive function after open heart surgery was examined using the Hasegawa's Dementia Scale (max. 30 points) in 47 patients with average age of 61.7 years. The patient's score was obtained preoperatively, and 1st and 7th postoperative days. Patients with postoperative scores above 24 points were classified as Non-Decline group, and those below 23 points as Decline group; comparative analyses were done on the relationship between various perioperative factors on both groups. Anesthesia was induced with low-dose fentanyl and isoflurane-nitrous oxide in oxygen. The average operating time was 331 minutes, and the cardiopulmonary bypass time was 108 minutes with normothermia. There were 31 Non-Decline cases, and 16 (34%) Decline cases on the 1st and 5 (10 %) on the 7th postoperative day. The largest decrease in the Decline group was seen on the 1st postoperative day, and there were 4 cases of dementia. The factors in which a significant difference was observed between the 2 groups were age, amount of transfusion, extubation time, preoperative complications and postoperative cardiac index, and these was not related to the operation or ECC time. Moreover there were high incidences of postoperative complications, especially LOS, in the Decline group. It was concluded that one of the largest contributing factors of abnormality of the cognitive function postoperatively was age, and other factors such as pre- and postoperative general conditions of the patients, especially cardiac function, might also be related. The length of hospitalization in the cases of decreased cognitive function was longer. PMID- 7474310 TI - [Postoperative management following open heart surgery by thoracic epidural anesthesia]. AB - The significance of thoracic epidural anesthesia as a postoperative analgesic sedative was evaluated. Ninety-seven open heart surgery patients with average age of 64 years, were classified into: Non-Epidural (N) group, Epidural with butorphanol (B) group and Epidural with morphine (M) group, in each of which the pulmonary, circulatory and sedative states were studied. Anesthesia was induced with low-dose fentanyl and isoflurane-nitrous oxide-oxygen, and epidural catheter was inserted through Th 7-10 on the day before the operation for Epidural groups. Butorphanol 0.5-1.0 mg was administrated in B group, and morphine 2-5 mg in M group. The Epidural groups required only small amount of fentanyl, 5 micrograms.kg-1, were extubated earlier compared to N group, and showed higher postoperative cardiac index. As for the respiratory state, although PaCO2 was slightly higher in the Epidural group, there was no difference in oxygenation. The incidence of administration of postoperative analgesic-sedative was extremely low in the Epidural especially in M group. As long as the indication is appropriately selected, the epidural anesthesia will not have negative effect on the postoperative pulmonary and circulatory state, allows for an earlier extubation and has excellent sedative effect. It is an effective method for patient management following open heart surgery. PMID- 7474311 TI - [Anesthetic management for laparoscopic cholecystectomy in a patient with dilated cardiomyopathy]. AB - A 55-year-old female with dilated cardiomyopathy was scheduled for laparoscopic cholecystectomy under general anesthesia. Pre-operative tests revealed abnormal ECG and hypokinesis of left ventricular wall motion on echocardiography. A detailed medical examination resulted in the diagnosis of dilated cardiomyopathy, although there was a very mild decrease in cardiac function. We determined that laparoscopic cholecystectomy could be safely carried out under general anesthesia if pneumoperitoneum pressure was kept low. Anesthesia was induced with diazepam 3.75 mg, fentanyl 75 micrograms, thiopental sodium 25 mg and vecuronium 6 mg. Anesthesia was maintained with isoflurane-nitrous oxide-oxygen. From the induction to the end of the operation, dopamine was infused. Increased heart rate and blood pressure were noted for a short time at the induction and at the time of first pneumoperitoneum. Thereafter, no considerable hemodynamic changes occurred. We therefore consider that laparoscopic cholecystectomy under low pressure pneumoperitoneum is appropriate for dilated cardiomyopathy when there is no accompanying heart failure or when only mildly decreased cardiac function exists, such as in this case. PMID- 7474312 TI - [Perioperative management of a patient with severe bronchial asthma attack]. AB - We report an anesthetic management for Miles' operation in a 50-year-old female who had frequent severe bronchial asthmatic attacks prior to surgery. Because the surgical field was in the lower abdomen, we selected spinal anesthesia combined with epidural anesthesia. For spinal anesthesia, 15 mg of hyperbaric tetracaine with epinephrine was used. Moreover, 3 ml of 2% mepivacaine with epinephrine was injected via an epidural catheter, and then analgesia up to Th 6 was obtained. In order to prevent an asthmatic attack during surgery, the following cares were taken: (1) administration of moisturized oxygen by nebulizer via a nasal canula, (2) intravenous administration of steroid, (3) addition of epinephrine to the local anesthetics, and (4) sedation by music. The surgery was completed safely without asthmatic attacks. Postoperatively, pain control with buprenorphine was managed with a continuous extradural infusion. The patient was discharged 50 days after surgery, and was in a favorable condition without bronchial asthmatic attacks. PMID- 7474313 TI - [Two patients who recovered fully from profound circulatory and respiratory failure after cardiac arrest]. AB - Two patients who were both 62-year-old males, also with various complications fully recovered from profound circulatory and respiratory failure. Both patients had experienced cardiac arrest one week or three months previously. Ischemic insult of brain produced from the prior cardiac arrest may be related to these excellent outcomes, because induction of tolerance to brain ischemia was demonstrated in animals which had undergone prior sublethal brain ischemia. PMID- 7474314 TI - [The effect of indomethacin suppository in preventing mesenteric traction syndrome]. AB - Mesenteric traction syndrome consists of cutaneous hyperemia with hypotension and tachycardia. NSAIDs could inhibit the phenomenon, but there are few reports about when to administer these drugs. In this study, we evaluated the effect of indomethacin on preventing mesenteric traction syndrome when administered preoperatively and just after induction of anesthesia. Thirty-six patients scheduled for abdominal hysterectomy were studied. Patients were randomized into three groups. Group C (n = 12); control, group T (n = 12); indomethacin 50 mg suppository just after induction of anesthesia, group P (n = 12); indomethacin 50 mg suppository about 90 min before incision. The effect of indomethacin was evaluated from the extent of cutaneous hyperemia. MTS was suppressed in group P, but not in group T (P < 0.05). We concluded that indomethacin suppository just after induction could not prevent mesenteric traction syndrome. PMID- 7474315 TI - [A case of atopic dermatitis treated with stellate ganglion block--the change of serum IgE and blood eosinophil levels]. AB - Stellate ganglion block (SGB) therapy was tried on a patient with severe adult type atopic dermatitis. SGB was performed 102 times in total and clinical symptoms improved gradually. Serum IgE and blood eosinophil levels, which correlate with disease severity, increased gradually following repeated SGB. After a series of SGB was stopped, clinical symptoms became worse and serum IgE and blood eosinophil levels increased again. We conclude that although SGB is presumably one of the effective therapies for severe adult type atopic dermatitis, it might be difficult to improve atopic constitution entirely because the patient was not cured completely following repeated SGB. PMID- 7474316 TI - [Convulsive seizure after craniotomy--comparison of isoflurane anesthesia and sevoflurane anesthesia]. AB - The incidence of postoperative convulsive seizure was compared retrospectively during either isoflurane or sevoflurane anesthesia in patients after craniotomy. Overall, 5 of a total of 125 patients suffered convulsive seizures after craniotomy: 2 (4%) of 45 patients with isoflurane anesthesia and 3 (4%) of 80 patients with sevoflurane anesthesia. No significant differences were observed between these two groups. The duration of anesthesia and duration of the MAC.h were similar between these two groups. The occurrence of postoperative convulsive seizure was not related with the inhalational anesthetic agents, the underlying disorders and patient characteristics. It is suggested that either isoflurane or sevoflurane anesthesia in neurosurgical patients may have no specific relationship with the postoperative convulsive seizures. PMID- 7474317 TI - [Anesthetic management of a patient with microcephaly associated with cerebral atrophy]. AB - We describe the management of a 35-year-old female with microcephaly, cerebral atrophy especially in frontal lobes, dwarfism and severe mental retardation. The patient underwent open reduction of fracture of the humerus neck suffered from traffic accident. Authors could not find any article on the anesthetic management of microcephaly but could locate some about mentally retarded patients claiming the difficulty in anesthetic induction as well as emergence and frequent incidence of convulsions. We anesthetized the patient with modified NLA with nitrous oxide-oxygen and interestingly very small doses of fentanyl (1.7 micrograms.kg-1) were required. The anesthetic and surgical courses were uneventful. The patient emerged immediately with intravenously administered naloxone and flumazenil with no excitement nor convulsion. We conclude that fentanyl is the drug of choice for mentally retarded patient with microcephaly and smaller doses per kg.weight are sufficient for orthopedic surgery. PMID- 7474318 TI - [Assessment of ventriculoarterial coupling in humans by transesophageal echocardiography and radial artery pressure tracing]. AB - To investigate coupling between the heart and arterial system in patients undergoing elective non-cardiac surgery, we determined both the ventricular elastance and the effective arterial elastance in two groups of subjects: normal group, 68 subjects without heart disease; and cardiac group, 33 subjects with heart disease. Left ventricular end-systolic (Ves) and end-diastolic (Ved) volumes were determined by transesophageal echocardiography. By assuming that left ventricular end-systolic pressure (Pes) is close to mean arterial blood pressure (MAP) and that x-axis intercept (Vo) is zero, the ventricular elastance (E'max) was approximated as MAP/Ves, and the effective arterial elastance (Ea) as MAP/(Ved-Ves). In 222 (74%) of the 299 measurements obtained in normal group, Ea/E'max was nearly 0.5, which is a condition for a maximal mechanical efficiency, while in 61 measurements (20%) Ea was almost equal to E'max (Ea/E'max = 1), which is a condition for maximal stroke work from a given end-diastolic volume. In contrast, in cardiac group, Ea/E'max was nearly 0.5 in 56 (41%) of the 137 measurements, while in 42 measurements (31%) Ea/E'max was nearly 1. In addition, although the value of Ea/E'max over 2, which represents severe heart failure, was not observed in normal group, Ea/E'max was over 2 in 10 measurements (7%) in cardiac group. Thus, the present results suggest that, as reported previously in awake patients, ventriculoarterial coupling is set toward higher left ventricular work efficiency in surgical patients without heart disease, whereas in patients with heart disease, ventricular and arterial properties are so matched as to maximize stroke work at the expense of the work efficiency. PMID- 7474319 TI - [The recent trend in preoperative fasting]. AB - It has been generally practiced that patients are restrained from any foods and water 4-6 hr prior to anesthesia. Recent trends of anesthesiologist, however, are to permit the patients to take fluid until 2-3 hr before inducing anesthesia, as described in the guidelines from Canadian Anaesthetists' Society in 1990, and in the authoritative textbooks of anesthesia. We compared effects of the preoperative fasting between age-matched groups of patients constrained to: 1) 6 hr absolute preoperative fasting, and 2) 6 hr fasting but free for taking clear fluid until 2 hr before operation. Residual gastric volume and pH immediately after inducing anesthesia were monitored. In the respective age groups from infant to the aged, no significant changes were observed between 1) and 2). Considering that light meal leaves the stomach in 1.5-3 hr and clear fluid almost immediately, these results accurately explain and support relevance of the current concept concerning the preoperative fasting. Furthermore, since actual incidence of aspiration pneumonia during anesthesia is sufficiently low, strict preoperative fasting may be unreasonable. We consider that reducing the fasting time may not increase the risk of pulmonary aspiration and recommend our practical method consisting of preoperative 6 hr withholding solid foods while allowing clear fluid ingestion until 2-3 hr before inducing anesthesia. PMID- 7474320 TI - [The utility of thermistor attached tracheal tube for pediatric patients in cardiac surgery]. AB - During cardiac surgery, esophageal, rectal, or bladder temperature is usually monitored as an index of core temperature; however, these methods are invasive, and often inconsistently reflect the central body temperature, especially in pediatric patients. The purpose of this study was to evaluate the utility of tracheal temperature monitoring during cardiac surgery in pediatric patients. Fifteen children (ages; 8 m.-7 yr.) undergoing cardiac surgery with cardio pulmonary bypass (CPB) were studied. Anesthesia was induced and maintained with high doses of fentanyl and intermittent doses of midazolam. After anesthetic induction, esophageal, rectal, bladder, tympanic, and forehead deep temperatures were monitored with a Core Temp Monitor (CTM-205, Terumo Co.). Simultaneously, the tracheal temperature was monitored with a specially made tracheal tube. A thermistor was attached by medical glue (Loctite Prism, Loctite Co.) at the anterior surface of a tracheal tube (Trachelon, Terumo Co.) without cuff (inside diameter 4.0-6.0 mm) where the tracheal tube tightly fits against the trachea. The inspired gas was warmed to 35 degrees C and humidified. During CPB, the blood temperature at the inlet to the patients was also recorded simultaneously. All indicated temperatures (Y) during CPB were analyzed for correlation with the blood temperature to the patients (X). The tracheal temperature had the highest correlation with the blood temperature from the patients (Y = 0.68X + 10.60, r = 0.89). There were also good correlations of the esophageal as well as bladder temperatures with blood temperature. There were no patients who suffered tracheal inflammation or laryngeal edema from the thermistor. Monitoring tracheal temperature is not only valuable for monitoring the core temperature, but also is convenient for pediatric patients in cardiac surgery. PMID- 7474321 TI - [The plasma concentration measurement of local anesthetics, thiobarbiturates and mexiletine by HPLC with automatic pre-treatment system]. AB - The concentrations of lidocaine, its metabolites (MEGX, GX), mepivacaine, bupivacaine, thiamylal, thiopental and mexiletine in plasma have been determined by HPLC method with automatic pre-treatment system. The drugs carried by pre treatment mobile phase are trapped by hydrophobic interaction on the pre treatment column (4 mm i.d. x 30 mm L., tetramethylolmethanetriacrylate resin packing). As sending analytical mobile phase which was conditioned with 0.1 M sodium phosphate buffer (pH 2.1) and acetonitrile by switching the flow changeover valve, the absorped drugs were eluted quickly and introduced to the analytical column (6 mm i.d x 150 mm L., octadecylsilan chemical bond sillica packing). On this column, the drugs were separated and analyzed, then measured by a UV spectrophotmetric detector. The time of switching valve, the wave length of detector for measurement and the buffer/acetonitrile ratio of analytical mobile phase were specific for the drugs, respectively. And high recovery and reproducibility were obtained for using auto-injector, so it has been possible to determine the concentrations of drugs in plasma with absolute calibration curves. PMID- 7474323 TI - [Lytic action of cefminox against slowly growing bacteria]. AB - Lytic action of cefminox (CMNX) against slowly growing Escherichia coli (E. coli) K-12 JE1011 was compared with that of the related cephamycin cefmetazole (CMZ) or ampicillin (ABPC). The growth rate in doubling time was 30 min. at 37 degrees C. Minimal concentrations of CMNX, CMZ and ABPC showing clear lysis were 0.39, 3.13 and 12.5 micrograms/ml, respectively, under these conditions. When E. coli was cultured at 12 degrees C, the doubling time was 720 and 816 min. which were 24 and 27.2 fold as slow as those at 37 degrees C, respectively. In 12 degrees C culture, the minimal concentrations showing clear lysis were 0.78 (2 fold at 37 degrees C), 25 (8 fold) and 100 micrograms/ml and higher (8 fold or more) with CMNX, CMZ and ABPC, respectively. Therefore, it appeared that the lytic activity of CMNX was not significantly affected by bacterial growth rate and was demonstrated effectively against slowly growing bacteria at a low concentration. We determined release activity of cell wall peptidoglycan fragments by 5 beta lactam antibiotics in addition to the above-mentioned 3 antibiotics. The results showed that the release activity was not proportional to the MIC and that of CMNX was the strongest among those antibiotics. The rapid and strong bactericidal action of CMNX against bacteria at the early stationary phase was suggested to result from the characteristic lytic action against slowly growing bacteria. PMID- 7474322 TI - [Use of Finapres during warming for postoperative patients]. AB - We used Finapres during rewarming after extracorporeal circulation in 16 patients who underwent cardiac surgery. In patients with passive rewarming there were no differences in blood pressure measured with a Finapres and that by an invasive monitor. On the other hand, in patients rewarmed with the aid of nitroglycerin, there were statistically significant differences between blood pressures measured with indirect and direct methods. PMID- 7474324 TI - [Antimicrobial activities of norfloxacin against clinical isolates from ocular infections]. AB - In order to evaluate antimicrobial activity of norfloxacin (NFLX), minimum inhibitory concentration (MICs) of NFLX and control drugs were determined against clinical isolates from ocular infections that were obtained in our laboratory from July, 1993 to December, 1994. The results are summarized as follows; 1. Compared to MIC distributions of NFLX against clinical isolates from ocular infections studied in 1986 and 1987, the MIC80 of NFLX against Corynebacterium spp., Enterobacter spp., Serratia spp., Burkholderia cepacia, Flavobacterium spp., Alcaligenes spp. increased 8 times. Almost all of NFLX-resistant strains among them were ofloxacin (OFLX)-resistant, new quinolones resistant strains, and a part of them were aminoglycosides, beta-lactams-resistant as well, thus all of these strains were multiple drug resistant. 2. MIC of NFLX against Pseudomonas aeruginosa were lower than that of OFLX. 3. NFLX showed strong antimicrobial activities against so-called "particular bacteria" including Staphylococcus aureus subsp. aureus, Moraxella spp., Haemophilus spp., and P. aeruginosa from ocular infections. And MIC80 of NFLX against these bacteria was 0.05-1.56 microgram/ml. We observed that NFLX eye drops was administered so that concentrations above the MIC against these clinical isolates were maintained. PMID- 7474325 TI - [Study of clinically isolated new quinolones-resistant Haemophilus influenzae. Part 1]. AB - A study was done to determine susceptibilities of Haemophilus influenzae that were obtained in our laboratory in 1994 to new quinolones (NQ) and other drugs. The results were as follows; 1. Among the 300 isolates, the detection frequency of NQ-resistant strains was 8.7% (26 strains), including isolates from chronic lower respiratory tract infections (22 strains) and those from middle meatus of nose (2 strains), etc. NQ-resistant strains were not isolated from children. 2. The cross resistance was studied for different NQs against NQ-resistant strains. Clavulanic acid/amoxicillin, cefteram, cefpodoxime, cefditoren, cefodizime (CDZM) and cefpirome showed strong antimicrobial activities against NQ-resistant strains. MIC90 of CEPs against all isolated strains including NQ-resistant strains and beta-lactamase producers was low. And the MIC90 of CDZM was < or = 0.025 microgram/ml, which was the lowest among all the antibiotics tested. 3. We found 47 strains (15.7%) of beta-lactamase producers among the 300 isolates, the frequency of beta-lactamase producing strains was high among strains obtained from children. PMID- 7474326 TI - [Susceptibilities of bacteria isolated from patients with respiratory infectious diseases to antibiotics (1991)]. AB - Isolated bacteria from respiratory tract infections were collected since 1981 in cooperation with institutions located throughout Japan, and have been investigated for their sensitivities to various antibacterial agents and antibiotics and reported by IKEMOTO, et al. Relationships between these isolates and backgrounds of the patients were also studied each year. These results are discussed in detail in this report. In 20 institutions around the entire Japan from October 1991 to September 1992, 631 strains of bacteria were isolated mainly from sputa of 529 patients with respiratory tract infections and tentatively determined to be etiological agents. MICs of various antibacterial agents and antibiotics against 96 strains of Staphylococcus aureus, 112 strains of Streptococcus pneumoniae, 111 strains of Haemophilus influenzae, 114 strains of Pseudomonas aeruginosa (non-mucoid), 41 strains of Moraxella subgenus Branhamella catarrhalis, 39 strains of Pseudomonas aeruginosa (mucoid), Klebsiella pneumoniae and some others, were determined, and the drug sensitivities of these strains were determined except for the strains that had been killed during transportation: 1. S. aureus. S. aureus strains for which MICs of methicillin were higher than 4 micrograms/ml (methicillin-resistant S. aureus) accounted for 58.3% and the frequency of the drug resistant bacteria increased over previous year's 42.5%. As shown by the MICs, arbekacin was active as vancomycin against all the strains on S. aureus. 2. S. pneumoniae: Benzylpenicillin among the penicillins showed a potent activity against S. pneumoniae. Cefuzonam, cefmenoxime, cefozopran and cefotaxime among the cephems showed excellent antimicrobial activities against S. pneumoniae. Imipenem; a penem antibiotic, showed the most potent activity with MIC80 of 0.03 micrograms/ml. 3. H. influenzae: Activities of all drugs were excellent against H. influenzae strains tested. Ampicillin showed MIC80 of 1 micrograms/ml against H. influenzae. Cefuzonam showed the most potent activity among cephems, it completely killed all bacteria at MIC 0.06 micrograms/ml. Cefotaxime and cefmenoxime showed next most potent activities with MIC80s of 0.06 micrograms/ml. The antimicrobial activity of ofloxacin was equivalent to those of cephems. 4. P. aeruginosa (mucoid). Ciprofloxacin and tobramycin showed the most potent activities against P. aeruginosa (mucoid), and their MIC80s were 4 micrograms/ml. 5. P. aeruginosa (non mucoid): Similarly, ciprofloxacin and tobramycin showed the most potent activities against P. aeruginosa (non-mucoid) with MIC80 of 2 micrograms/ml. Comparing to activities against P. aeruginosa (mucoid), all the drugs tested showed lower activities against P. aeruginosa (non-mucoid). 6. K. pneumoniae: The activities of all drugs except for penicillins were very high against K. pneumoniae.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 7474328 TI - [Significant role of azole antifungal agents: focused on itraconazole]. PMID- 7474327 TI - [Pharmacokinetic study of sparfloxacin in patients receiving regular hemodialysis]. AB - We investigated the in vivo kinetics of sparfloxacin (SPFX), an oral quinolone, in the hemodialytic patients. SPFX was orally administrated in a single dose of 200 mg to each of five hemodialytic patients, on a day that they were not receiving hemodialysis therapy. After dosing, the blood samples were collected periodically at 2, 4, 6, 8, 12, 24, 25, 27, 29, 48, 72 hours. Concentration of the unchanged SPFX in the plasma samples were measured by HPLC. The peak plasma levels of SPFX in the hemodialytic patients did not differ greatly from patients with renal failure (5 < Ccr<20 ml/min). Average T 1/2 beta of SPFX was 25.7 hours, and it was prolonged compared to the patients with renal failure (who were not receiving hemodialysis) and a group of young healthy volunteers. Based on these results, we believe those hemodialytic patients may be administrated with the drug once every other day, as long as normal doses are used. PMID- 7474329 TI - [Pharmacokinetic and clinical studies with azithromycin (fine granule) in the pediatric field. Pediatric Study Group of Azithromycin]. AB - Azithromycin (AZM) in 10% fine granules, a newly developed azalide antibiotic, was administered at a standard dose of 10 mg/kg once daily for 3 to 5 days (89.5% received 3 day administration) to children with infectious diseases and the efficacy and the safety of AZM were investigated. In addition AZM concentrations were determined in blood samples from 18 patients and in urine samples from 17 patients to examine o pharmacokinetic characteristics of AZM. 1. Absorption and excretion: Cmax's in 16 patients who received 10 mg/kg and 2 patients who received 20 mg/kg were 0.29 +/- 0.24 micrograms/ml and 0.75 micrograms/ml, respectively, while T 1/2's were 42.0 +/- 11.8 hours for the former and 51.3 hours for the latter. AUC(0 to approximately infinity)'s were 10.72 +/- 5.00 micrograms x hr/ml in the former and 28.83 micrograms x hr/ml in the latter. Urinary concentrations of AZM peaked at 48 to 72 hours after the administration of 10 mg/kg AZM in 14 patients, while it peaked at 24 to 48 hours in the patients who received 20 mg/kg. Urinary recovery rates in the first 120 hours after the start were 9.1 +/- 2.6% for 10 mg/kg and 10.8 +/- 3.4% for 20mg/kg. 2. Clinical efficacy: The study received 619 entries and 564 cases were evaluated for drug efficacy. The remaining were not evaluated because of dropout or exclusion. The efficacy rate, combining both "Excellent" and "Good" cases was 94.3% in 246 cases where pathogens were identified, classified as Group A. The efficacy rate was 90.7% for the remaining 321 cases, classified as Group B, where causative pathogens were unidentified. The difference between the two groups was no statistical significance. The combined efficacy rate was 92.2%. For the 116 cases where the patients had failed to respond to previous chemotherapies instituted for 3 days or longer, the efficacy rate for AZM was 94.0%. 3. Adverse reactions and abnormal laboratory tests: Incidents of diarrhea, soft stool, skin rashes, or vomiting were found in 15 patients (2.5%) of 596 cases eligible for evaluation. These reactions, however, were all transient and mild to moderate in severity in the 15 patients including 4 patients for whom the treatment was discontinued, all resolved in time. Abnormal changes in laboratory tests were found as follows: decrease in WBC in 23 patients (5.6%), increase in eosinophils in 28 (7.1%), increase in platelet count in 2 (0.5%), decrease in platelet count in 1 (0.3%), elevation of GOT in 3 (0.8%), and elevation of GPT in 6 (1.6%).(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 7474330 TI - [Pharmacokinetic and clinical studies with azithromycin (capsule) in the pediatric field. Pediatric Study Group of Azithromycin]. AB - Azithromycin (AZM) in 100 mg capsules, a newly developed azalide antibiotic, was administered at a standard dose of 10 mg/kg once daily for 3 to 5 days (89.9% received 3 day administration) to children with infectious diseases and the efficacy and the safety of AZM were investigated. In addition, AZM concentrations were determined in blood samples from 9 patients and in urine samples from 12 patients to examine pharmacokinetic characteristics of AZM. 1. Absorption and excretion: Cmax was 0.45 +/- 0.28 micrograms/ml, T 1/2 was 52.7 +/- 20.2 hours, and AUC(0 approximately to infinity) was 12.09 +/- 4.93 micrograms.hr/ml in the 9 patients each of whom received 8.5 to 14.3 mg/kg AZM. Urinary concentrations of AZM peaked at 48 to 72 hours after the administration of 8.5 to 14.7 mg/kg AZM in 12 patients and the average urinary recovery rate in 120 hours was 7.3 +/- 2.8%. 2. Clinical efficacy: The study received 139 entries and 119 cases were evaluated for drug efficacy. The remaining were not evaluated because of dropout or exclusion. The efficacy rate combining both "Excellent" and "Good" cases, was 100% for 40 cases in which pathogens were identified, classified as Group A. The efficacy rate was 97.5% for the remaining 79 cases, classified as Group B, where causative pathogens were unidentified. The difference between the two groups was no statistical significance. The combined efficacy rate was 98.3%. For the 31 cases where the patients had failed to respond to the previous chemotherapies instituted for 3 days or longer, the efficacy rate for AZM was 93.5%. 3. Adverse reactions and abnormal laboratory tests: 8 incidents of diarrhea, skin rashes, urticaria, or vomiting were found in 7 patients (5.4%) of 130 cases eligible for evaluation. These reactions, however, were all transient and mild to moderate in severity in the 7 patients including 2 patients for whom the treatment was discontinued, all resolved in time. Abnormal changes in laboratory tests were found as follows: decrease in WBC in 10 patients (9.3%), an increase in eosinophils in 12 (11.4%), an increase in platelet count in 1 (1.0%), an elevation of GOT in 3 (3.1%), an elevation of GPT in 6 (6.2%), and an elevation of LDH in 1 (1.1%). The abnormalities were transient and did not require particular intervention. Moreover, none of the patients indicated clinical signs associated with the abnormal changes of laboratory tests.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7474332 TI - [Effects of azithromycin on fecal flora of healthy adult volunteers]. AB - New macrolide antibiotic, azithromycin (AZM), was administered to six healthy male volunteers and its effects on their intestinal microflora were investigated. Each volunteer was given 500 mg of AZM orally, once a day for 3 consecutive days. Stool samples were obtained from them prior to the medication and 1, 7, 14 and 28 days after the third day the medication. A slight decrease in the total aerobic bacterial count was observed. Also, several species of anaerobic bacteria showed slight decreases through the 14th day post medication. Individual variances were observed, however. A marked decrease in the number of Bifidobacterium was found for each of the volunteers. Clostridium difficile was detected from one of the volunteers on the 28th day post medication without diarrhea related symptoms. PMID- 7474333 TI - [Comparative studies on activities of antimicrobial agents against causative organisms isolated from urinary tract infections (1989). I. Susceptibility distribution]. AB - The frequencies of isolation and susceptibilities to antimicrobial agents were investigated on 1,032 bacterial strains isolated from patients with urinary tract infections in 10 hospitals during the period of June 1989 to May 1990. Of the above total bacterial isolates, Gram-positive bacteria accounted for 30.8% and a majority of them were Enterococcus faecalis. Gram-negative bacteria accounted for 69.2% and most of them were Escherichia coli. 1. Enterococcus faecalis Imipenem (IPM) showed the highest activity against E. faecalis isolated from patients with urinary tract infections. The followings, ampicillin (ABPC) and vancomycin (VCM) showed potent activities, with the MIC90S of 2 micrograms/ml. Piperacillin (PIPC), minocycline (MINO) and chloramphenicol (CP) were also active with the MIC90S of 8 micrograms/ml. The others were not so active with the MIC90S of 32 micrograms/ml or above. 2. Staphylococcus aureus VCM showed the highest activity against S. aureus with MIC90 of 1 microgram/ml. Dicloxacillin (MDIPC) and arbekacin (ABK) were active with the MIC90S of 2 micrograms/ml. MINO showed the MIC90 of 4 micrograms/ml. All other agents except ciprofloxacin (CPFX) showed lower activity. 3. Staphylococcus epidermidis MINO showed the highest activity against S. epidermidis. Its MIC90 was 0.25 microgram/ml. The followings, ABK and VCM were also active with the MIC90S of 0.5 microgram/ml, 2 micrograms/ml, respectively. The others except CPFX were not so active. 4. Coagulase-negative staphylococci (CNS) Most of the agents were active against CNS. IPM, ABK and MINO showed the highest activities with the MIC90S of 0.125 microgram/ml or below. MDIPC, cefazolin (CEZ), cefotiam (CTM) and VCM were also active with the MIC90S of 1 microgram/ml. Clindamycin (CLDM) showed lower activity, with the MIC90 of 128 micrograms/ml. 5. Streptococcus agalactiae CEZ, cefuzonam (CZON), IPM and CLDM showed the potent activity, all strains were inhibited at the MIC of 0.125 microgram/ml or below. The followings, cefmenoxime (CMX) and erythromycin (EM) were active with the MIC90S of 0.125 microgram/ml or below. PIPC and VCM were also active with the MIC90S 0.25 microgram/ml and 0.5 microgram/ml, respectively Amikacin (AMK) showed lower activity. Escherichia coli IPM, CTM, flomoxef (FMOX), CMX, carumonam (CRMN), norfloxacin (NFLX), ofloxacin (OFLX) and CPFX showed the highest activities against E. coli. The MIC90S of them were .125 microgram/ml or below. ceftazidime (CAZ) and CZON were also active with the MIC90S of 0.25 microgram/ml. Penicillins except mecillinam (MPC) were not so active showing the MIC90S of 32 micrograms/ml or above. 7. Klebsiella pneuminiae FMOX, CMX, cefixime (CFIX), IPM, CRMN and NFLX showed the highest activities against K. pneumoniae.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 7474334 TI - [Comparative studies on activities of antimicrobial agents against causative organisms isolated from urinary tract infections (1989). II. Background of patients]. AB - Clinical background was investigated on 1,197 bacterial strains isolated from patients with urinary tract infections (UTIs) in 10 hospitals during the period from June, 1989 to May, 1990. 1. Distribution of sex and age A majority of female patients with UTIs accounted for a twenties and thirties. The proportion of male patients with UTI has increased with age, accounting for majority in patients 70 years or older. 2. Distribution of infection types and age Most cases among twenties or thirties were uncomplicated UTIs. Fifties and older cases were most frequent in complicated UTIs. 3. Distribution of isolated bacteria and age Escherichia coli was most frequent on the whole, followed by Enterococcus faecalis, Pseudomonas aeruginosa, Staphylococcus spp. and Klebsiella spp.. E. coli had declined with age, and on the other hand, E. faecalis and P. aeruginosa had increased with age. 4. Administration of antibiotics and pathogens isolated from UTIs. In uncomplicated UTIs, pathogens, after administration of antibiotics, isolated from patients have obviously decreased from 439 to 4. Before an administration of antibiotics, E. coli was mainly detected, but after an administration of antibiotics, E. coli has decreased into two strains. After administration of antibiotics, pathogens of complicated UTIs, without indwelling catheter, have decreased from 324 to 19: E. coli has decreased from 86 to 2, but E. faecalis, Staphylococcus spp., Enterobacter spp. and P. aeruginosa have increased. Pathogens of complicated UTIs with indwelling catheter, have decreased from 156 to 14 after administration of antibiotics, and mainly isolated pathogens were E. faecalis and P. aeruginosa. PMID- 7474331 TI - [Comparative clinical study of azithromycin with tosufloxacin tosilate in the treatment of acute odontogenic infection]. AB - To objectively assess azithromycin (AZM) for its clinical efficacy, safety and usefulness in the treatment of acute odontogenic infections (periodontitis, pericoronitis and osteitis of the jaw), a double-blind, randomized, multi-center trial was conducted in which tosufloxacin tosilate (TFLX) was used as the control drug. AZM was administered to 90 patients at a once-daily 500 mg dose for 3 days, while TFLX was given to 90 patients at a 150 mg t.i.d. dose for 7 days. 1. The clinical efficacy rates calculated according to evaluation at an endpoint set on the 3rd day of treatment by a committee of experts were 85.9% (73/85) in the AZM group and 78.9% (71/90) in the TFLX group. No statistically significant difference between the treatment groups was detected, and clinical equivalence was verified (p = 0.002). 2. The clinical efficacy rates according to evaluations made by investigators at the end-of-tail point was 87.1% (74/85) in the AZM group and 73.3% (66/90) in the TFLX group. The efficacy rate in the AZM group was higher than that in the TFLX group, and the difference was statistically significant (p = 0.006). 3. The bacteriological elimination rate in the AZM group was 97.5% (39/40) and that in the TFLX group was 85.7% (30/35), but the difference was deemed statistically not significant. 4. Adverse reactions were observed in 11 of 88 cases (12.5%) in the AZM group and 5 of 90 cases (5.6%) in the TFLX group. Six of 85 cases (7.1%) in the AZM group and 5 of 85 cases (5.9%) in the TFLX group showed laboratory abnormalities. However, neither adverse reactions nor laboratory abnormalities showed any differences in statistical significance between the treatment groups. 5. The safety rates, expressed as percentages of cases with no adverse events and no laboratory abnormalities, was 84.1% (74/88) in the AZM group and 90.0% (81/90) in the TFLX group. The difference between the two groups was found to be statistically insignificant. 6. The usefulness rates, the ratio of cases rated as either "Very useful" or "Useful", was 83.9% (73/87) in the AZM group, and it was statistically higher (p = 0.025) than 72.2% (65/90) obtained for TFLX group. Judging from the above results, it has been concluded that AZM is as useful as TFLX in the treatment of acute dental infections. PMID- 7474335 TI - [Comparative studies on activities of antimicrobial agents against causative organisms isolated from urinary tract infections (1989). III. Secular changes in susceptibility]. AB - Susceptibilities of Citrobacter spp., Enterobacter spp., Escherichia coli, Klebsiella spp., Proteus mirabilis, Pseudomonas aeruginosa, Serratia spp. isolated from patients with urinary tract infections (UTIs) in 10 hospitals during June 1989 to May 1990 to various antimicrobial agents were compared with those in the same period of previous years according to a classification, uncomplicated UTIs, complicated UTIs without indwelling catheter, and complicated UTIs with indwelling catheter. As for Citrobacter spp., P. mirabilis and Serratia spp., which were detected very few in 1989, their susceptibilities were not observed an obvious change. As for Enterobacter spp., the susceptible strains to flomoxef, cefixime, cefuzonam and ceftazidime increased in complicated UTIs with indwelling catheter. The susceptibilities of E. coli to penicillins increased slightly in complicated UTIs with indwelling catheter. Against Klebsiella spp., a good activity of minocycline or cephems was found. The susceptibilities of P. aeruginosa to ciprofloxacin and new quinolones increased in uncomplicated UTIs. These data should be considered in clinical treatment of various urinary tract infections. PMID- 7474336 TI - [Frequency of clinical isolation of glucose non-fermentative gram-negative rods and their susceptibilities to antibacterial agents]. AB - A comparison was made for frequencies of isolation o glucose non-fermentative Gram-negative rods ((G)NF-GNR) from clinical specimens during a period from July, 1986 to June, 1987 (the first period) and that from January, 1994 to December, 1994 (the second period). Also, minimum inhibitory concentrations of principal drugs were determined against these isolates. The obtained results are summarized as follows: 1. Thirty four (34) species of (G)NF-GNR were found from 35,200 clinical specimens in the two periods. Numbers of strains of (G)NF-GNR obtained were 4,575 during the first period and 4,704 during the second period, thus no significant difference existed in numbers of strains isolated in the two periods. 2. Among the 34 species to which the 4,704 strains were classified into, Pseudomonas aeruginosa comprised 68.4%, Stenotrophomonas maltophilia 6.9%, Acinetobacter baumannii 5.6%, Burkholderia cepacia 3.1%, Acinetobacter Iwoffii 2.6%, Alcaligenes xylosoxidans subsp. xylosoxidans 2.4%, Flavobacterium indologenes 1.7%, Pseudomonas putida 1.1%, Acinetobacter junii 1.1% and Moraxella subgenus Moraxella lacunata 0.9%. When these frequencies of isolation were compared with those in the first period, it was found that B. cepacia decreased significantly (P < 0.01) and that S. maltophilia increased significantly (P < 0.001). 3. MIC determinations revealed multiple drug resistance strains in many different species of bacteria. Minocycline, however, were active against many such strains, and ofloxacin was found to have strong antibacterial activity against some strains. PMID- 7474337 TI - [Pharmacokinetics of balofloxacin in the intraocular tissues of pigmented rabbits]. AB - We performed the pharmacokinetics of balofloxacin in plasma and intraocular tissues in pigmented rabbits upon its oral administration. The penetrations of balofloxacin into plasma and aqueous humor of the anterior chamber reached their peaks of 6.46 micrograms/ml and 0.70 microgram/ml, respectively, and the ratio of drug concentration in aqueous humor to that of plasma was 0.12 at 1 hour after an oral administration of 20 mg/kg. Concentrations in plasma and aqueous humor of the anterior chamber reached 0.27 microgram/ml and 0.18 microgram/ml, respectively, at The T 1/2 (alpha) and T 1/2 (beta) of the drug in aqueous humor were longer than those in plasma The T 1/2 (alpha) and T 1/2 (beta) of the drug in aqueous humor were longer than those in plasma at a dose of 20 mg/kg. The concentrations in iris and ciliary body were much higher than in any other intraocular tissue in 24 hours after single administration of 20 mg/kg, and those concentrations remained at high levels for a long time. PMID- 7474339 TI - [Shibasaburo Kitasato's discovery of plague bacillus and its historical background--on the occasion of the centennial anniversary]. PMID- 7474338 TI - [Pathogenicity of Shigella flexneri and its regulation]. PMID- 7474340 TI - [Pathogenicity of Yersinia pestis: microbiological and molecular aspect]. PMID- 7474341 TI - [Molecular biology of Candida albicans]. PMID- 7474343 TI - [Q fever and Coxiella burnetii]. PMID- 7474342 TI - [Attempts to elucidate the reasons why mycobacterial infections are generally intractable]. PMID- 7474344 TI - [Macrolides, the attractive antibiotics--from a structural and functional aspect]. PMID- 7474345 TI - [Regulation of the toxin production in Clostridium perfringens]. PMID- 7474346 TI - [Comparative biology of Legionella species--with special reference to host parasite relationships]. PMID- 7474347 TI - [Principles and practice of control of hospital infection]. PMID- 7474348 TI - [Helicobacter pylori and gastroduodenal diseases]. PMID- 7474350 TI - [Are cytokines factors for host defense against infection?]. PMID- 7474349 TI - [Influenza-bacillus: who discovered the organism?]. PMID- 7474351 TI - [Validly published pathogenic bacterial names and new biosafety level of National Institute of Health]. PMID- 7474352 TI - [Lists of bacterial names validated in 1994]. PMID- 7474353 TI - [Cattle botulism in Australia]. PMID- 7474354 TI - [Causative agent of the so-called "light disease of shrimps" is luminescent Vibrio cholerae non-O1]. AB - A number of luminous fresh-water shrimps were found in a fish preserve in Lake Biwa, Shiga Prefecture, in the middle of July, 1994, and most of them died within several hours after collection (the so-called "light disease of shrimp"). Four luminous organisms were isolated from a dead shrimp. Although the phenotypic properties of these strains were similar to those of V. cholerae or V. mimicus, a representative strain, 838-94, was shown to have a high level (79%) of DNA homology with V. cholerae type strain, ATCC 14035 and a low level (45%) of relatedness to V. mimicus type strain, ATCC 33653. Therefore, these isolates were identified as V. cholerae. The four strains fell into serogroup O28 of V. cholerae. On the other hand, none of the isolates had CT nor NAG-ST genes. The results obtained herein clearly demonstrate that these organisms isolated from luminous shrimps are luminescent V. cholerae serogroup O28. PMID- 7474355 TI - [Autobacteriographic studies on the distribution and localization of Staphylococcus aureus in mice]. AB - Autobacteriography was proposed as a bacteriological method to follow distribution and localization of bacteria in experimentally infected animals. Infectious organisms were restricted to rifampicin-resistant strains to prevent contamination during autobacteriography. Mice were infected with Staphylococcus aureus Smith diffuse type RFPr (rifampicin-resistant) by the intravenous route and frozen at various intervals after infection. Whole body sections (40-microns thick) of the mice were transferred onto selective agar medium containing rifampicin to incubate at 37 degrees C. On day 1 after infection, dense colonies of the infecting organism on the sections were distributed in the whole body. On day 3, few organisms were detected in the liver and many were observed in the spleen, kidney and intestinal tract. On days 7, 14 and 21, the organisms in the liver and spleen disappeared, and those in the kidney and intestine remained. The remaining infectious organisms were demonstrated in the kidney and intestinal tract by autobacteriography of mice infected with S. aureus Smith compact type RFPr. By cultivation of the homogenate of the gastrointestinal tissues and their contents, the infectious organisms were detected mainly in the lower small intestines, cecum and large intestines. PMID- 7474356 TI - [Characterization by RFLP of DNAs from Campylobacter jejuni Lior serotype reference strains and clinical isolates and detection of C. jejuni by DNA-probe]. AB - The Lior serotype reference strains and clinical isolates of Campylobacter jejuni were compared in the restriction fragment length polymorphism (RFLP) pattern to distinguish C. jejuni strains. These reference strains showed RFLP patterns different from one to another, while the patterns of some isolates were not coincident with those of the same serotype reference strains. Furthermore, we tried to hybridize HindIII-digested fragments from these strains with the DNA probe encoding the 46-kDa protein of C. jejuni by Southern and slot blottings. The 1.8-kbp fragments from all strains of C. jejuni hybridized with this probe, but those from other species of Campylobacter or enterobacteria did not. These results indicate that the Lior serotype is unrelated with the RFLP pattern of DNA of C. jejuni strains, but the DNA probe is useful to detect C. jejuni. PMID- 7474357 TI - Usefulness of hyperventilation thallium-201 single photon emission computed tomography for the diagnosis of vasospastic angina. AB - To establish a safe and sensitive diagnostic procedure for detecting coronary vasospasm, we utilized 201-thallium myocardial SPECT combined with hyperventilation (HV-SPECT) in 29 patients with vasospastic angina (VAP) and 11 controls. Twenty-five of 29 patients with VAP and 5 of 11 controls developed transient perfusion defects on HV-SPECT, resulting in a sensitivity and specificity calculated at 86% and 55%, respectively. Overall accuracy in identifying corresponding vessels with coronary vasospasm, respectively. Coronary vasospasm tended to be identified more accurately in the left anterior descending branch and the right coronary artery than in the circumflex branch (75%, 71% and 50%, respectively). The hyperventilation test induced ischemic ECG changes in 11 of 29 patients with VAP, yielding a sensitivity of 38%. Analyzing the washout rate of HV-SPECT in patients with VAP, both the extent and severity scores of patients with ischemic ECG changes were larger than those of patients without. No serious complications occurred during HV-SPECT. In conclusion, HV-SPECT was a safe and sensitive procedure as a primary diagnostic approach for VAP. From the results of washout analysis, HV-SPECT could detect more mild myocardial ischemia than could the ECG, and seemed quite useful especially for detecting coronary vasospasm accompanied by minimal ischemic ECG changes. PMID- 7474358 TI - Plasma levels of endogenous opioid peptides in patients with acute myocardial infarction. AB - There is substantial evidence that cardiac opioid receptors are activated during arrhythmias induced by administration of opioid peptides or myocardial ischemia, supporting the hypothesis that endogenous opioid peptides (EOP) are involved in myocardial infarction. This prospective clinical trial is designed to determine whether the ischemia-induced arrhythmias and extent of the infarct are related to the release of the EOP beta-endorphin in patients with acute myocardial infarction. Two groups were included in the study, patients with acute myocardial infarction, and healthy volunteers who served as controls. The results indicate that, compared to the controls, there was augmentation of ischemic arrhythmias and ischemic damage as assessed by serum creatine kinase activity, accompanied by an elevated level of beta-endorphin, in patients with acute myocardial infarction. The above data strongly indicate that EOP are indeed involved in the pathophysiology of myocardial infarction, and suggest these peptides have an important role in ischemic heart disease. PMID- 7474359 TI - Glomerular hemodynamics during supraventricular tachycardia. AB - Supraventricular tachycardia (SVT) was induced in 7 patients by programmed cardiac stimulation via an esophageal lead. Blood pressure, renal function, and hormonal factors were measured before, during, and after SVT. The glomerular filtration rate increased during SVT, while renal blood flow did not change. The parameters of glomerular hemodynamics were calculated according to the method of Gomez. Glomerular pressure was 64 +/- 3 (mean +/- SE) mmHg before SVT, and rose significantly (p < 0.01) to 76 +/- 5 during SVT. This rise in glomerular pressure was associated with a decrease in afferent vascular resistance (from 3355 +/- 610 to 1770 +/- 517 dynes x sec/cm5, p < 0.05) and an increase in efferent vascular resistance (from 3726 +/- 758 to 4814 +/- 780 dynes x sec/cm5, p < 0.05). Since atrial natriuretic peptide (ANP) increased during SVT (from 40 +/- 15 to 208 +/- 72 pg/ml, p < 0.01), these changes in glomerular hemodynamics may be attributed to the physiologic action of ANP. Despite the changes in glomerular hemodynamics during SVT, natriuresis appeared after SVT and not during SVT. This suggests that natriuresis accompanying SVT could not be attributed to the changes in glomerular hemodynamics. PMID- 7474361 TI - Effects of trandolapril on cardiac angiotensin I converting enzyme activity in rats with chronic heart failure following myocardial infarction. AB - The effects of long-term treatment with a novel angiotensin I converting enzyme (ACE) inhibitor, trandolapril, on ACE activity and cardiac function in rats with chronic heart failure (CHF) were examined and compared with those of captopril and enalapril. Left coronary artery ligation of rats resulted in decreases in mean arterial pressure, left ventricular systolic pressure, dP/dt, cardiac output and stroke volume indices, and increases in left ventricular end-diastolic pressure and systemic vascular resistance 12 weeks after the operation. A significant increase in ACE activity of the myocardium, but not that of serum or other tissues, was detected in the CHF rat 12 weeks after the operation. Oral treatment with ACE inhibitors (10 mg/kg/day captopril, 10 mg/kg/day enalapril or 3 mg/kg/day trandolapril) from the 2nd to 12th week, attenuated the changes in cardiac output and stroke volume indices, left ventricular end-diastolic pressure and systemic vascular resistance of the CHF rat. Treatment also attenuated the increase in the cardiac. ACE activity of CHF rats. A close relationship between the decrease in cardiac output index and the increase in cardiac ACE activity was detected. The results suggest that trandolapril, like other ACE inhibitors, exerts a beneficial effect on cardiac function in the CHF rat and that one of the mechanisms for this effect is attenuation of elevated cardiac ACE activity. PMID- 7474360 TI - Effects of atrioventricular intervals on right atrial monophasic action potential duration during pacing or reciprocating tachycardia. AB - We evaluated the relationship between changes in the atrioventricular (AV) intervals and right atrial monophasic action potential duration at 90% repolarization (RA-MAPD90) in humans. In 4 patients, RA-MAPD90 was measured during AV reciprocating tachycardia (AVRT, n = 2) or AV node reentrant tachycardia (AVNRT, n = 2) and atrial or ventricular pacing at similar cycle lengths. In 10 patients, RA-MAPD90 was measured during continuous atrioventricular sequential pacing at AV intervals of 0, 50, 100, 150, 200, 250, 300, 350 msec for 90 sec at a cycle length of 333-400 msec. In patients with AVRT, the RA-MAPD90 during tachycardia was shorter than RA-MAPD90 during RA or RV pacing. In patients with AVNRT, RA-MAPD90 during tachycardia was similar or longer than RA-MAPD90 during RA or RV pacing. As the AV interval was increased from 0 msec to 150 msec, the peak right atrial pressure decreased from 8-12 mmHg to 2-3 mmHg. RA-MAPD90 decreased to its minimal value at an AV interval of 150 msec. The maximal difference in the RA-MAPD90 obtained by changing the AV interval was 22.5 +/- 3.0 msec. We conclude that 1) a very short AV or VA interval causes an increase in RA-MAPD90 which correlates with an increase in RA pressure, and 2) RA-MAPD90 during supraventricular tachycardia is different from RA-MAPD90 during atrial or ventricular pacing. Thus, a contraction-excitation feedback mechanism may exist in the human atrium. PMID- 7474362 TI - Effects of ajmaline on non-sodium ionic currents in guinea pig ventricular myocytes. AB - The lack of currently available data stimulated us to investigate the electrophysiological effects of ajmaline, a classical class Ia antiarrhythmic agent, on various currents responsible for the action potential plateau and repolarization phases. The whole cell patch clamp recording technique was applied to guinea pig ventricular myocytes. Ajmaline suppressed the Ca2+ current (Ica) in a dose-dependent manner (Kd = 1.2 x 10(-5) M) without affecting the steady-state inactivation kinetics and the voltage dependency of the current-voltage relationship. Ajmaline inhibited the inward portion of the inward rectifying K+ current (IKl). Ajmaline decreased the delayed rectifier K+ current (IK) without altering the activation or deactivation time courses. All these inhibitory effects of ajmaline prolonged the action potential duration in a dose dependent manner. The inhibitory actions of ajmaline on the action potential upstroke and various currents responsible for the plateau and repolarization may contribute to the observed suppression of depolarization-induced abnormal automaticities by this agent. PMID- 7474363 TI - Nicorandil suppresses a hump on the monophasic action potential and torsade de pointes in a patient with idiopathic long QT syndrome. AB - The patient was a 71-year-old female with Torsade de Pointes (TdP) associated with idiopathic long QT syndrome. TdP and polymorphic nonsustained VT were frequently observed at bedside and an electrophysiologic study was performed. The QT (and QTU) interval was abnormally prolonged, and alteration of the QT interval was also recorded on the electrocardiogram. Monophasic action potential (MAP) from the right ventricle showed a hump on the falling limb of the MAP following a long RR interval of more than 1.0 sec. Intravenous administration of nicorandil (2 mg) resulted in disappearance of the hump, and ventricular arrhythmia was no longer observed. The QT interval at a PP interval of 720 msec was slightly shortened. She was treated with a DDD-pacemaker and given nicorandil. No recurrence of TdP was observed during the follow-up period of 8 months. This drug might be effective in patients with idiopathic long QT syndrome. PMID- 7474364 TI - Sustained-release procainamide-induced reversible granulocytopenia after myocardial infarction. AB - A 71-year-old man with paroxysmal atrial fibrillation who had a previous anterior myocardial infarction exhibited granulocytopenia 8 days following the administration of oral sustained-release procainamide (750 mg/day). The plasma concentrations of procainamide and N-acetyl procainamide were at subtherapeutic levels. Discontinuation of procainamide led to complete recovery. A bone marrow aspiration showed slight hypoplasia with normocellular marrow. Lupus erythematosus (LE) and antinuclear antibody (ANA) tests were negative. The frequency and relationship of granulocytopenia caused by sustained-release procainamide in patients with tachyarrhythmias are briefly discussed, and prior reported cases are reviewed. Precautionary measures for the early recognition of this grave hazard in exposed patients are advocated. The physician should be aware of this complication before in initiating treatment with this drug. PMID- 7474365 TI - [Past, present and future of enzyme immunoassay]. AB - Enzyme immunoassay (EIA) is one of the most popular non-isotopic immunoassay methods, with various clinical applications. After radioimmunoassay was developed by Yalow and Berson, its competitive binding principle was combined with enzyme labeling techniques and its application has been expanded by developing various procedures. Theoretically, EIA is more sensitive than RIA, being able to measure levels as minute as some attomole levels. Automated enzyme immunoassay systems have further improved its intra-laboratory variation, but relatively large variations among different reagent kits must be improved. Highly sensitive EIA systems should allow wider clinical applications. PMID- 7474366 TI - [Various types of immunoassay]. AB - Five types of immunoassay, enzyme immunoassay (EIA), radioimmunoassay (RIA), fluoroimmunoassay (FIA), chemiluminescent immunoassay (CLIA) and counting immunoassay (CIA), are generally used. Radioimmunoassay was first developed but it needs specific facilities and the half life of radioisotope is not long. Enzyme immunoassay is at present most popular in Japan. The goal of development of immunoassay is improvement of sensitivity and automation. In order to improve sensitivity of an assay, immunoenzymometric assay (IEMA), which is a non competitive method, was developed and fluorescent materials and chemiluminescent materials are used for the detection of enzyme activity. Automation of immunoenzymometric assay could be done with solid phase antibody method. PMID- 7474367 TI - [Competitive immunoassay and immunometric immunoassay]. AB - Radioimmunoassays are simple and quite useful, being easily applied on a commercially serviceable basis. Furthermore radioimmunoassays are extremely sensitive and quite precise. Two site immunoradiometric assay in which one antibody was immobilized and another labelled with 125I becomes gradually popular, due to its higher sensitivity and the development of monoclonal antibody production by the hybridoma technique. Alternative analytical methods other than radioimmunoassays (radioimmunoassay, immunoradiometric assay) have been developed, in which the alternative labels to substitute for radioisotopes are enzymes, luminescent compounds and fluorescent probes. PMID- 7474368 TI - [An immunoblot technique]. AB - After a protein is separated by various types of gel electrophoresis, it is transferred onto a nitrocellulose membrane, where a specified protein is detected through immunological reactions. This procedure, called an immunoblot technique, combines gel electrophoresis, which is capable of separating materials and immunological detection with specificity and high sensitivity. It has recently been used widely as a useful method for analyzing proteins in the fields of medicine, biochemistry, molecular biology and human genetics. The aim of this technique is to transfer a target protein, which is present in the gel, as efficiently as possible onto a membrane so that it can react uniquely on the membrane. To achieve this end, it is necessary to select the following; 1) a transfer system, 2) a transfer membrane, 3) a transfer buffer and additive, 4) optimal transfer conditions, and 5) a specific and highly sensitive means of immunological detection, to be combined with the technique. Since the immunoblot technique is, in principle, capable of analyzing all types of proteins as long as there is an antibody specific to the target protein, it can be used widely to determine the physiopathology of human proteins. PMID- 7474369 TI - [Dynamics of antigen-antibody interaction--an introduction]. AB - Dynamic or thermodynamic theory for antigen-antibody interaction was discussed in this chapter. The reaction between antibody and antigen may be simplistically described by the law of mass action. The strength of a reaction between monovalent antigen and monovalent antibody is referred to as affinity. Antibody affinity is influenced by many factors, which include the degree of the stereochemical fit between the combing site and the epitope and the size of the region. In this paper, I would like to discuss how to detect and quantitatively express about antigen-antibody interaction using by several theories. PMID- 7474370 TI - [Enzyme kinetics for enzyme immunoassay]. AB - The rate of reaction involving an enzyme is greatly influenced by temperature, pH, concentration of the substrate, and a number of other factors. Therefore, an accurate measurement of enzyme-catalyzed reaction is important for enzyme immunoassay. The substrate concentration needed for half-maximum velocity (1/2 Vmax) is called the Km value (Michaelis constant) and is expressed in units of substrate concentration (moles per liter or M). Km may be considered an approximate measure of affinity of an enzyme for its substrate: the lower the Km, the higher is the affinity. At times, optimum conditions cannot be used, and compromises in optimum assay conditions must be made. PMID- 7474371 TI - [The effects of solid phase preparation on sensitivity of enzyme immunoassay]. AB - The preparation of solid phase immobilized antibody or antigen was physical adsorption or chemical binding using coupling reagent such as glutalaldehyde etc. The concentration of antibody or antigen on solid phase was effective to sensitivity of enzyme immunoassay. When the nonspecific binding of enzyme conjugate to solid phase was minimized, the sensitivity of enzyme immunoassay will be succeed in higher. PMID- 7474372 TI - [Factors that affect analytical results in an enzyme immunoassay]. AB - Factors that affect an enzyme immunoassay were reviewed from the practical point for clinical laboratory uses: these include prozone effect or hook effect, affinity and avidity, cross-reactivity and specificity of the antibody, and other non-specific factors such as pH, temperature, rheumatoid factor, complement, heterophile antibody and others. Precise mechanisms still remained unclear for these factors involved in non-specific reactions in the assay. PMID- 7474373 TI - [General properties and kinds of antibodies for enzyme immunoassay- characteristics and advantages of polyclonal and monoclonal antibodies]. AB - General properties and advantages between monoclonal and polyclonal antibodies are described with regard to their specificity, cross reaction, practical consideration, affinity and isotype. Although monoclonal antibodies are superior in specificity, they have less affinity than those in polyclonal antibodies. Since monoclonal antibodies usually react with a single epitope, they sometimes show a particular characteristic to an antigen. It should be emphasized that investigation of required properties is important to obtain the best combination of antibodies for immunoassay purposes. PMID- 7474374 TI - [Properties of enzymes for enzyme immunoassay]. AB - Enzyme selection is very important to establish enzyme immunoassay system. The appropriate enzyme selected to label an immunochemical reagent must have certain qualities as follows: 1) highly purified and high specific activity, 2) inexpensive, 3) stable during conjugation and for a long time in a proper storage condition, 4) easily conjugated, measured, and applied to highly sensitive assay. Several enzymes fulfill most of the above requirements and have been successfully used in enzyme immunoassay. These are horseradish peroxidase, alkaline phosphatase, beta-galactosidase, glucose oxidase, glucose-6-phosphate dehydrogenase, etc. As for enzyme activity measuring procedures, chemiluminescence and bioluminescence assays generally exhibit more sensitive detection limits than fluorescent or colorimetric assays. In the future, ultrasensitive luminescent assays and production of excellent recombinant enzymes are expected. PMID- 7474375 TI - [Enzyme labeling methods and it's specificities]. AB - Enzyme labeled antigen for use in ELISA of Hapten (steroids, prostanoid, carbohydrate, nucleic acid, peptide, herbicide, insecticide and antibiotic) have usually been prepared by condensation of carboxy group of hapten with amino groups of lysine residue in enzyme. The horseradish peroxidase (HRP) is best suitable as labeling enzyme, therefore it is small molecular, and substrate turnover is much higher compared to the other enzymes. The mixed anhydride and carbodiimide methods have mainly been used the preparation of hapten conjugate BSA, but not satisfactory for enzyme labeling. The N-hydroxysuccinimide ester (NHS: active ester) method is satisfactory with respect to reproducibility and sensitivity. The sensitivity is related to the bridging phenomenon: One of the disadvantages of the homologous labels is that the antibody shows an affinity not only for the Hapten but also for the bridge which connects the Hapten to carrier protein. We have developed a sensitive bridge heterologous EIA for progesterone (P) using geometrical isomers of P-3 (E/Z) (O-carboxymethyl) oxime-N hydroxysuccinimide esters [ef Ab of P-3 (E) CMO-BSA/P-3 (Z) CMO-HRP]3). The sensitivity of heterologous proved to be higher than a homologous EIA or a conventional RIA. It seem like that a 1:1 steroid-enzyme conjugate is suitable for obtaining a high sensitivity. The avidin-biotin (AB) system provides great versatility, since by conjugation with an appropriate label, the AB assay can be used with any chosen detector. Furthermore, IgG can be labels with biotin without significantly influencing their immunological activity.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7474376 TI - [Characteristics of enzyme substrate used for enzyme immunoassay]. AB - Enzyme immunoassay in measured its enzyme activity as a signal. This signal detection method have been progressing from colorimetric to fluorometric determination, and most recently amongst for more sensitive substrate chemiluminescent determination is popular. The methods, colorimetric determination is recognized to be less sensitive. We confirmed that the alkaline phosphatase labeled enzyme, cycling method significantly increased its sensitivity by changing the substrate. If the minimum detection limit is not sensitive enough by colorimetric determination, fluorometric is useful, and chemiluminescence can be applied when ultra sensitive method is required. We briefly described the characteristics of the main substrates which may very the sensitivity in these assays. PMID- 7474377 TI - [Characteristics of various labeled chemiluminescent compounds]. AB - A chemiluminescence has a diverse range of analytical applications and an attractive analytical method for the third generation analysis because a high sensitive luminometer is now available for use with chemiluminescent reactions. A combination of chemiluminescence and a high technology provides a high sensitive assay system for biological substances such as peptide, hormones, antigen and antibody. Characteristics of luminol-dioxetane- and acridium-derivatives as a labeled chemiluminescent compound are reviewed for applications on CLIA and CLEIA. PMID- 7474379 TI - [Kinds and characteristics of solid-phase enzyme immunoassay]. AB - Technology with solid-phase enzyme immunoassay was markedly progressed. Kinds of solid-phase are as follows; (1) tube, (2) beads, (3) latex particle, (4) microtiter plate, (5) ferrite particle, (6) membrane. Recently, application of avidin/biotin system to the tube method was accomplished. As one of the B/F separation with beads method, spin washing method was also developed. Automated open system with the beads method was also developed. Microtiter plate using sponges as the solid-phase was routinely used for allergy tests (CAP-RAST). Multi Access Open System (MAOS), applicable for various kinds of ELISA kit, will appear in Japan near future. Latex particle with ferrite plating and ferrite particle with glass coating were also used as the solid-phase of EIA. HCV-RIBA II and III systems using nitro-cellulose membrane as solid-phase were also described. PMID- 7474378 TI - [B/F separation systems in enzyme immunoassay]. AB - Methods for bound/free (B/F) separation in enzyme immunoassay are reviewed. In liquid-phase enzyme immunoassays, double antibody methods are used in small laboratories. In solid-phase enzyme immunoassay, which is a more popular and convenient method, antibody (antigen) is usually bound to the support by physical adsorption, and B/F separation is accomplished by washing the solid phase. Covalent binding of proteins, DNA, and oligosaccharides is also possible by using chemically modified supports. Indirect binding of antibody (antigen) to the support mediated by avidin-biotin methods has several merits in comparison with direct methods and double antibody methods. The immune complex transfer method has been used for ultrasensitive assays. The use of magnetic particles as a support can accelerate the immune complex formation, and is becoming more popular in clinical laboratories. A rapid sandwich enzyme immunoassay, using a porous filter as a support, has been developed for dry chemistry. PMID- 7474380 TI - [B/F non-separative method]. AB - In homogeneous enzyme immunoassay without requirement of separation of free and bound label, the concentration of substances in samples can be detected by measurement of enzyme activity in the mixtures of samples (antigen) and reagents (labeled antigens antibodies, and enzyme substrates). The methods described herein offer the virtues of simplicity and rapidity by use of general-purpose automated clinical analyzer. The concentrations of some drugs, hormones, antibiotics, and very small amount of certain proteins are now detected by the routine examination in these techniques employing commercial diagnostic kits offered by several companies. In this manuscript, we attempted to give an overview of the principle of various methods by use of commercial diagnostic kit applied in routine clinical examinations. PMID- 7474381 TI - [The assay of enzyme activity in EIA--colorimetric, fluorometric and luminometric method]. AB - The method of enzyme activity related in enzyme immunoassay was described. The detection methods were colorimetric, fluorometric and chemi- or bioluminometric assay for enzyme, and the sensitivity of three detection methods were compared. The order of detection limit was chemi- or bioluminometry > fluorometry > colorimetry. But it is important that the selection of detection method of enzyme will make fit on your desirable sensitivity of analysis. PMID- 7474382 TI - [Enzyme amplification in enzyme immunoassay]. AB - In enzyme immunoassay, the catalytic activity is determined by photometric assay of the substrate, and in a broad sense, it also includes fluorometric and chemiluminescent assays. Since enzyme immunoassay excells in specificity and detectability, it is widely applied to clinical tests which analyze various specimens. Here, we describe some of enzyme amplification methods including cyclic reactions, avidin-biotin reactions and enhanced chemiluminescent enzyme immunoassays. These methods have resulted in highly sensitive immunoassays, the level of which ranges from a attomole (1 x 10(-18) mol) to a zeptomole (1 x 10( 21) mol). PMID- 7474383 TI - [Automated analyzer of enzyme immunoassay]. AB - Automated analyzers for enzyme immunoassay can be classified by several points of view: the kind of labeled antibodies or enzymes, detection methods, the number of tests per unit time, analytical time and speed per run. In practice, it is important for us consider the several points such as detection limits, the number of tests per unit time, analytical range, and precision. Most of the automated analyzers on the market can randomly access and measure samples. I will describe the recent advance of automated analyzers reviewing their labeling antibodies and enzymes, the detection methods, the number of test per unit time and analytical time and speed per test. PMID- 7474384 TI - [Quality control of enzyme immunoassay--assurance and control of analytical sensitivity, accuracy and precision]. AB - Although the basic concept for quality control of enzyme immunoassay (EIA) is the same as that of quality control of the usual chemical analysis, immunological analytic methods including enzyme immunoassay have some specific features depending on the method as compared with the chemical method. In addition to the usual QC method, therefore, it is necessary to add some QC techniques to control the variables derived from the method. In accuracy control of EIA, standard materials, reagents and analytical equipment will be the major sources of error. Points to be considered in accuracy control are described. In precision control of EIA, between-run precision is generally larger than within-run precision. In consideration of the characteristics, mean-Rs-R control chart method and Z-V control chart method appropriate for immunological method are described. Analytical sensitivity expressed as detectability and detection limits is very important in EIA. A procedure to determine the detection limits is described in detail. Between reagent lot difference in immunological reagent is considerably larger than that of chemistry reagent and a shift of the point on the control chart is not adequate to assess the reliability of the patient data. Therefore, an evaluation method of the between reagent lot difference is described. General procedures of precision control are not described here because the procedures are well known. PMID- 7474386 TI - [Calibration and standardization in enzyme immunoassay]. AB - Calibration is performed using multipoints of the calibrator in enzyme immunoassay (EIA). Calibration curves in EIA are frequently represented by non linearity. Therefore, calibration in EIA is fitted with suitable functions. These functions are demonstrated with logistic curve, logit-log conversion, spleen function, etc. Consequently, the calibration procedure is indicated for the manufacturer's instruction, and also the calibrator is prepared as a commercial available reagent kit. To minimize EIA measurements among laboratories, and among methods, standardization with a common suitable reference material is needed. Reference materials for EIA, international standards are prepared by the expert committee of WHO. These standards must be transferred to national standards, manufacturer's standards and calibrators of commercial available reagent kits. Finally, the measured value from the routine method is maintained with commutability and traceability. PMID- 7474385 TI - [Problems related to determination of reference interval and clinical decision limit in enzyme immunoassay]. AB - It is generally difficult to determine reference interval in enzyme immunoassay (EIA) at an individual laboratory because of many sources of variations for analytes measured by EIA, low analytical precision and high reagent cost of EIA. Large within-kit variations can be reduced by strict control of assay procedures so that the reference interval can be determined jointly and transferred among users of the same kit. To set clinical decision limits (CDL) it is necessary to consider low frequency of abnormality in tumor markers and hormones when they are measured for case finding. The CDL which minimize an expected cost can be determined when costs for false positive/negative results are given together with pretest probability, sensitivity and specificity. PMID- 7474387 TI - [Cause and countermeasure for appearance of false positivity or false negativity in enzyme immunoassay (EIA)]. AB - The appearance of false positivity or false negativity in EIA (enzyme immunoassay) brings often trouble of judgment for diagnosis or monitoring of disease. Most of the appearance can be found out by the dilution test of test sample or the recovery test by addition of standard antigen to test sample. It was pointed out that the assay conditions, the assay method, the dose of enzyme labeled antibody and the enzyme employed in EIA system might affect the appearance, and the autoantibody to antibody employed in the system, the enzyme and the antibody of idiotype in test sample might become factors of the appearance. Furthermore, the countermeasure for the appearance was concretely described in the paper. PMID- 7474388 TI - [Present status and future of enzyme immunoassay--hormone]. AB - Recently various non-radioisotopic immunoassays have been developed instead of radioisotopic assay and widely used in the laboratory. Enzyme immunoassay (EIA) is most popular in use including chemiluminescent enzyme immunoassay (CLEIA). At present few hormones (thyroid related and gonadotropic hormones) are measured by EIA with semi or full automatic analysers and satisfactory specificity and sensitivity were already obtained. However, most hormones are still measured by RIA or IRMA. EIA for these hormones should be developed. Standardization of reference interval and calibrators would also be needed for the compatibility between different assays. PMID- 7474389 TI - [Isozyme analysis by enzyme immunoassay (EIA)--clinical significance and future prospect]. AB - Serum isozymes are widely used for estimating the origin or the degree of injury. The significance and future prospect of enzyme immunoassay (EIA) for the determination of isozymes are summarized in this paper. EIAs were developed for isozymes, e.g., unstable or labile, and indistinguishable by electrophoresis or ion-exchange column chromatography EIA can measure isozyme fragments with no catalytic activity. The catalytic activity of isoenzymes released from tissues disappears immediately but immunologically active isozymes or its fragment may remain in the blood for a longer period. The determination of isozymes by EIA provides an accurate mass, released from the injury. The combination assay of catalytic activity with isozyme mass by EIA may open a new approach for the disease. PMID- 7474390 TI - [Enzyme immunoassay (EIA) as an assay system of tumor markers]. AB - To detect a small amount of tumor marker in the serum, the development of a sensitive assay system has been tried. In the beginning radioimmunoassay was mainly employed. But the use of radioisotope has been undesirable because the special facilities to treat radioactive substance or to discard it are required. For this reason the various assay systems without the radioisotope have been developed. Among them EIA system is in the mainstream, especially for the determination of CEA and AFP. Recent analysis shows the increasing trend of the employment of EIA for the determination of other tumor markers. In EIA system it is critical to keep the regulation of reaction time to be obtain the accurate value. Autoassay system is favorable for this problem. It is also another problem that discrepancies of assayed value are seem among the different assay kits. In the clinical use of tumor marker the comparison of the serial data, even if they are obtained from the different hospitals, is important. So overcoming this problem is desirable. PMID- 7474391 TI - [Enzyme immunoassay of substances related to allergy]. AB - The substances related to allergy that we can measure by EIA were described. Allergic reactions begin from the reaction of an antigen and IgE on the membrane surface of basophils and mast cells. The chemical mediators are released from the cells activated by the reaction of antigen and IgE. The chemical mediators proceed to the reactions of focal or total body inflammation. Measurements of serum IgE, chemical mediators, and cytokines are important parameters for the diagnosis and therapy of allergic disease. Antigen analyses are possible by application of EIA. It is also possible to apply EIA in the prevention of allergic diseases. PMID- 7474392 TI - [Enzyme immunoassay of blood coagulation and fibrinolysis factors]. AB - Various types of immunoassays are at present applied to inspection items in clinical aspect of blood coagulation and fibrinolysis, in which EIA is acting as one of the major methods. Commercial EIA kits, available in Japan, can be itemized into categories of blood coagulation, fibrinolysis, endothelial cell injury, and platelet activation systems, along with other types of immunoassays and enzymatic activity assays, itemized for comparison. Assay methods suitable for an objective antigen should be selected, based on its biological, biochemical characteristics, such as plasma concentrations, and even clinical demands. The matters to be solved range over making reliable standard materials, using monoclonal antibody to detect absolute concentrates of specific antigen, and then further discovery of molecular markers that specifically reflect coagulative or fibrinolytic activation, leading to thrombotic or bleeding tendency. PMID- 7474393 TI - [ELISA for diagnosis of infections by viruses]. AB - The assay used most widely to detect or diagnose virus infection, especially infection of blood borne viruses e.g. HBV, HCV, HIV and HTLV, is the enzyme linked immunosorbent assay (ELISA), whose sensitivity and practicability have rendered it the most common primary screening assay. ELISA can be mass screening used automatic or semiautomatic machines. ELISAs can be indirect assay, competition assays or sandwich assays. In indirect and sandwich assays, the development of color indicates the presence of antigen or antibody, whereas in competition assays the absence of color development signifies a positive reaction. Alkaline phosphatase and horseradish peroxidase the most commonly used enzymes, are associated with their respective substrates, usually p-nitro-phenyl phosphate and hydrogen peroxide. The ELISA for antigen detection is used polyclonal antibody or monoclonal antibodies. The other hand, the ELISA for antibody screening is used whole virus, synthetic peptides or recombinant antigens. PMID- 7474394 TI - [Enzyme-immunoassay for autoantibodies using recombinant antigens]. AB - The Enzyme-immunoassay (EIA) is known as safe and sensitive quantitative-method to detect autoantibodies. However, it has not been used routinely because of difficulty to prepare a large amount of native autoantigens. Many investigators have established EIA system using recombinant proteins to overcome this problem. And some of them have been useful for the diagnosis of disease and evaluation of disease activity. However, some sera positive for autoantibodies to nuclear antigens such as U1 RNP and Sm, do not react with recombinant proteins because those autoantibodies often recognize the conformational profile of the autoantigen. Therefore, using both double immunodiffusion and EIA is recommended to diagnose the patient with connective tissue disease and to evaluate the disease activities, and may clarify the clinical significance of EIA using recombinant antigens. PMID- 7474395 TI - [Enzyme immunoassay for cytokines]. AB - Cytokines produced by different types of cells function by cell to cell interaction both in vitro and in vivo. Over thirty different molecules react in almost all physiological phenomena, such as in the fields of immunology, hematology, endocrinology, neurology, cardiology and nephrology. Previous observations indicate that the abnormal production of cytokines plays a part of pathogenesis of diseases. Therefore, it is necessary to know the amount of cytokines in the biological specimens for recognizing the pathogenic significance of cytokines. In this paper, we show the EIA-Kits of cytokines which are purchased. And we demonstrate the methods, advantages and disadvantages in the different enzyme immunoassays (EIA, DELFIA and Chemiluminescen-EIA) of interleukin 6 which we extensively investigated from basic to clinical experiments may be routinely used in the clinical examination in the near future. PMID- 7474396 TI - [Detection of bacterial protein toxins by a bead-ELISA]. AB - A highly sensitive bead-enzyme-linked immunosorbent assay to detect bacterial protein toxins was developed. Fab' of anti-toxin IgG was conjugated with horseradish peroxidase by the maleimide method and tetramethylbenzidine was used as a substrate. As the solid phase, a 6.5 mm diameter polystyrene bead was used and this was coated with the anti-toxin IgG. The sensitivities of the bead-ELISA for various bacterial protein toxins were as follows: less than 40 pg/ml for cholera enterotoxin (CT), less than 20 pg/ml for VT1 and less than 6 pg/ml for VT2 of enterohemorrhagic Escherichia coli. The bead ELISA was evaluated for direct detection of CT from stool specimens of patients with acute secretory diarrhea. Of the 75 stool samples examined, 59 yielded biochemically and serologically confirmed strains of Vibrio cholerae O1. The bead ELISA was positive for CT in stool supernatants in 50 (84.7%) of the 59 samples from which V. cholerae O1 was isolated. In addition, the bead ELISA was positive for three stool specimens which were negative by culture. These data indicate that the bead ELISA is a sensitive and simple method for direct detection of CT in nonsterile stool samples. PMID- 7474397 TI - [Enzyme immunoassay of drug in blood (serum)]. AB - Antiepileptic drugs, cardiac glycosides (digoxin, etc.), theophylline, antiarrhythmic agents (quinidine, procainamide, lidocaine, etc.) and aminoglycosides have narrow therapeutic range and large interindividual variations have been reported for their pharmacokinetics. Therefore, to obtain the appropriate therapeutic effects without undesirable effects, monitorings of above drug concentrations are useful (therapeutic drug monitoring). There are many assay methods (high-performance liquid chromatography, immunoassay, enzyme immunoassay, etc.) to determine drugs in serum. In those assays various enzyme immunoassays, enzyme-multiplied immunoassay technique (EMIT), enzyme-linked immunosorbent assay (ELISA), apoenzyme reactivation immunoassay (ARIS), radial partition immunoassay (RPIA), etc., are developed and widely used in many clinical laboratories. These assay systems offer convenience and great rapidity of assay and satisfactory accuracy and reproducibility but lack a little specificity of antibody. PMID- 7474398 TI - [Enzyme immunoassay of pesticide residues in foods]. AB - Analytical techniques for pesticide residues in food include gas chromatography (GC), gas chromatography/mass spectrometry (GC-MS) and high performance liquid chromatography (HPLC). These procedures are time consuming, cumbersome and require specialized instrumentation. A number of enzyme immunoassay methods for pesticide have appeared in recent years. In addition, nowadays a lot of immunoassay kits for various kinds of pesticide are in the market. The assay offers alternative means that is more sensitive, faster and cost-effective than previous methods. However, it is subject to problems that the assay does not suit multi-residue analysis and has possibility to encounter interferences which would affect the assay precision in case of some crops. Assuming that researcher knows which pesticide were applied to the analyzed crops beforehand, the pesticide would be detected easily by immunoassay. Also, when suitable sample kit for analyzed pesticide is available, the assay could be an excellent screening tool for large number of food samples. In conclusion, it will be necessary to consider analytical purpose to confirm utility of immunoassay when applying to pesticide analysis. PMID- 7474399 TI - [Detection of food additives by enzyme immunoassay]. AB - The analysis of synthesized food additives is generally performed by chromatography or spectrophotometry. However, the analytical procedures for natural food additives have been little reported so far because they are difficult to analyse chemically. We have attempted to apply enzyme immunoassay (EIA) to the analysis of natural food additives. Hen egg white lysozyme, as a food preservative, was determined by the competitive EIA, using mouse anti-HEL ascites. Carminic acid (CA), which is the main component of cochineal color, was determined by the competitive EIA, using monoclonal anti-CA antibody. Phycocyanin, which is the main component of spirulina color, was determined by the avidin-biotin sandwich EIA, using double monoclonal anti-phycocyanin antibodies. PMID- 7474400 TI - [Application of enzyme-linked immunosorbent assay (ELISA) to the medico-legal identification]. AB - Medico-legal identification of saliva stains and bloodstains was performed by an enzyme-linked immunosorbent assay (ELISA) using a horseradish peroxidase conjugate in combination with the use of monoclonal antibodies. Activity of alpha amylase in the stains was measured for an identification of saliva using an anti human amylase antibody, and secretory IgA was detected for a species identification using an anti-IgA antibody. ABO and Lewis blood group antigens were detected using anti-A, anti-B, anti-H, anti-Lea and anti-Leb antibodies. After the solubilization of ABH antigens in blood stains with octyl-beta-D glucose solution, ABO typing has been performed using fully automated system composed of a unit for blood group reaction based on ELISA and a unit for reading of result based on photometric analysis. PMID- 7474401 TI - [Towards immuno-modulation through the molecular mechanism of lymphocyte activation]. AB - Recognition of the antigen/MHC complex by the T cell receptor (TCR)-CD3 complex in T cells triggers early activation events such as tyrosine phosphorylation, phosphatidylinositol turnover, intracellular Ca2+ mobilization or activation of protein kinases, and finally exhibits effector functions such as lymphokine secretion by helper T cells or cytotoxicity by killer T cells as late activation events. Several key molecules have been shown to engage in these signaling cascades. In addition to the TCR-CD3 molecules, other surface molecules such as CD28 or LFA-1 contribute to the regulation of T cell activation as a co stimulator. Growing knowledge about the downstream of antigen recognition is promoting the attempt to modulate the signal transduction by specific drugs, mAbs, altered peptides or cytokines. Further investigations on the molecular mechanism of T cell activation will provide clinical successes to control immune responses. PMID- 7474402 TI - [New anti-HIV drug which binds the oligosaccharides of HIV envelope glycoprotein]. AB - The virion surface of the human immunodeficiency virus (HIV-1) is covered with an envelope glycoprotein gp120. Study of the oligosaccharide structures of gp120 suggests that the high mannose type of oligosaccharides are essential for HIV-1 infection. Pradimicin A, an antifungal antibiotic isolated from Actinomadura hibisca, and the derivative BMY-28864 have the ability to inhibit HIV-1 infection in vitro. The inhibitory effect was suppressed by addition of high mannose type oligosaccharides of gp120. BMY-28864 bound directly to gp120, mannose-BSA, and neoglycolipids containing high mannose type oligosaccharides but not to natural mammalian glycoproteins. The binding was Ca2+ dependent and was inhibited by mannose. BMY-28864 is a unique carbohydrate-binding antibiotic which has never been reported. It is possible to block HIV-1 infection by targeting oligosaccharide chains of the envelope glycoprotein. PMID- 7474403 TI - Infrequent mutations of the WT1 gene in primary cancers of the adult urinary tract. AB - Polymerase chain reaction-single strand conformation polymorphism (PCR-SSCP) analysis of all of the ten exons of the WT1 gene and restriction fragment length polymorphism (RFLP) analysis of the WT1 locus were performed on primary urinary tract cancers: seven renal pelvic cancers, one ureteral cancer, 11 bladder cancers, and 22 renal cell cancers. Four human bladder cancer cell lines (T24, JTC30, JTC32, and HUB41) and three human prostate cancer cell lines (PC-3, DU145, and LNCaP) were also examined. None of the primary cancers showed any apparent mutations of the gene, whereas one base substitution of exon 5 was found in DU 145 and gross alteration of the gene was recognized in HUB41. Heterozygosity of polymorphic exon 7 was retained in all of the 12 informative cases, and none of 10 informative cases showed loss of heterozygosity at the WT1 locus in RFLP analysis. It is concluded that mutations of the WT1 gene may not be involved in the formation of malignant tumors of the adult urinary tract. PMID- 7474404 TI - Significance of serum neuron-specific enolase as a predictor of relapse of small cell lung cancer. AB - We conducted a prospective study to evaluate the significance of serum neuron specific enolase (NSE) as a predictor of relapse of small cell lung cancer (SCLC). Patients entered into the study were drawn from those who had shown a complete or partial response to first-line chemotherapy with a concurrent decline in the NSE level to less than 10 ng/ml. When the serum NSE level increased to more than 15 ng/ml, the patient was restaged on the basis of clinical, radiological, and bronchoscopic examinations. During the period from August 1988 to December 1990, 57 patients with SCLC were enrolled and followed up until May 1992. Of these patients, 45 had clinical relapses, and 14 (31%) of them showed a clear elevation of the serum NSE level prior to the clinical recognition of relapse. Although one false-positive case was noted, this involved only a transient elevation of the NSE level. In patients who showed increased NSE levels, the relapses occurred in more difficult to detect silent sites such as the adrenal gland, liver, and deep lymph nodes. In addition, the percentage of patients demonstrating high NSE levels who were able to benefit from salvage chemotherapy was higher than for those who did not (RHO < 0.05). Our results indicate that serial NSE measurements are useful for the early prediction of SCLC relapse and should help to facilitate early administration of salvage chemotherapy for affected patients. PMID- 7474405 TI - Role of endoscopic retrograde cholangiopancreatography in laparoscopic cholecystectomy. AB - Endoscopic retrograde cholangiopancreatography was performed before laparoscopic cholecystectomy in 312 patients, with successful results in 287 patients (92.0%). Common bile duct stones were diagnosed in 26 of the 287 patients (9.1%). In eight of these patients, the diagnosis had not been suspected from the clinical or laboratory data or results of ultrasonography. Endoscopic sphincterotomy with stone removal was successful in all 23 patients who underwent this procedure. Anatomical variations were detected in 7 patients (2.4%), including origin of the cystic duct from the right hepatic duct and the accessory bile duct, and anomalous arrangement of the pancreaticobiliary ducts and choledochocele. Pancreaticobiliary malignancies were also diagnosed in 3 patients. The morbidity rate with diagnostic ERCP was only 0.7%. Preoperative ERCP proved highly useful and sale for determining biliary anatomy, and for detecting unsuspected stones or malignancies. PMID- 7474407 TI - Relation of family history of cancer and environmental factors to the risk of colorectal cancer: a case-control study. AB - The relation of a family history of cancer and environmental factors to colorectal cancer was investigated in a case-control study conducted from 1992 to 1994 at 10 medical institutions in Japan using a self-administered questionnaire, and 363 cases of colorectal cancer were compared with 363 controls matched for sex and age. A family history of colorectal cancer was positively associated with colon cancer (odds ratio (OR) = 2.0, 95% confidence interval (CI) 1.03-3.87) and rectal cancer (OR = 2.1 CI 0.94-4.48), but a family history of other cancers did not increase the risk. The proportion of patients with a family history of colorectal cancer within first-degree relatives was 12.4%--appreciably higher than figures previously reported in Japan. On the other hand, the incidence of hereditary non-polyposis colorectal cancer was 1.4%, and lower than previous estimates. Among dietary factors, a western-style diet significantly increased the risk of both colon and and rectal cancer (OR = 2.3 CI 1.30-3.88 and OR = 2.1 CI 1.26-3.63, respectively). Consumption of rice was protective against both colon and rectal cancer (OR = 0.5 CI 0.31-0.82 and OR = 0.3 CI 18-0.65, respectively). Animal meat, oily food, fish, vegetables and fruit were shown to affect the risk, but no statistically significant correlation was found. Among other factors, constipation increased the risk of colon cancer (OR = 2.0 CI 1.02 3.76) and consumption of coffee raised the risk of rectal cancer (OR = 1.7 CI 1.07-2.82). Our findings suggest that a family history of colorectal cancer is an important risk factor for this disease, and does not contradict the hypothesis that the risk of colorectal cancer in Japan may be influenced by westernization of lifestyle. However, we were unable to find conclusive evidence that familial clustering of this disease is strongly affected by environmental factors or genetic diseases such as hereditary non-polyposis colorectal cancer. PMID- 7474408 TI - Intensive chemotherapy for anaplastic thyroid carcinoma: combination of cisplatin, doxorubicin, etoposide and peplomycin with granulocyte granulocyte colony-stimulating factor support. Chemotherapy Committee, The Japanese Society of Thyroid Surgery. AB - The Japanese Society of Thyroid Surgery undertook a pilot study of treatment for anaplastic thyroid carcinoma in a cooperative setting. The treatment consisted of cisplatin 40 mg/m2 drip intravenous infusion (div), day 1, adriamycin 60 mg/m2 iv, day 1, etoposide 100 mg/m2/day div, days 1-3, peplomycin 5 mg/body/day sc, days 1-5 and granulocyte colony-stimulating factor (G-CSF) 2 micrograms/kg/day sc, days 6-14. This was scheduled to be repeated every 3 weeks. Local radiation therapy was added for patients in whom it was indicated. A total of 17 patients (mean age, 66 yr) were enrolled. Ten patients had advanced disease with measurable lesions and 2 patients experienced partial remission lasting 2 and 3 months, respectively. Six of 7 patients were treated with the same modality of treatment as an adjuvant. Three died of progressive disease after 3-7 months and three others have survived for 3-11 months. The toxicities of the chemotherapy were mainly bone marrow suppression, despite G-CSF support. Transient liver dysfunction was also noticed. These results indicate that this combined a treatment can be given to patients with acceptable toxicity. The degree of leukopenia was greater than expected, partly due to the advanced age of the patients and the low dose of G-CSF. In addition, 8 available thyroid specimens were examined for the mdr 1 gene and P-glycoprotein, but all were negative. Further study of anaplastic thyroid carcinoma by this cooperative group will be carried out. PMID- 7474406 TI - Malignant tumors of the nasal cavity: review of a 60-case series. AB - Sixty cases of primary malignant tumor of the nasal cavity treated in our hospital between 1962 and 1993 were reviewed. Males were affected 2.8 times more frequently than females. The age at the first consultation ranged from 11 to 92 years, with a mean of 55.1 years. The peak distribution was seen in the 6th decade. Twenty-six cases were epithelial malignancies (squamous cell carcinoma 15; adenocarcinoma 3; adenoid cystic carcinoma 3; undifferentiated carcinoma 3; mucoepidermoid carcinoma 1; malignant mixed tumor 1), while 34 cases were non epithelial malignancies (malignant melanoma 14; malignant lymphoma 14; plasmacytoma 3; olfactory neuroblastoma 2; rhabdomyosarcoma 1). The most common symptom on presentation was nasal obstruction (66.7%), followed by epistaxis (55.0%). The first recurrence was local in 19 patients, whereas cervical lymph node metastasis occurred in three patients, bone metastasis in two, intraperitoneal metastasis in two, and brain metastasis in one. The overall five year cumulative survival rate was 48.0%. The five-year survival rates for squamous cell carcinoma, malignant melanoma, and malignant lymphoma were 57.0%, 31.0%, and 40.0%, respectively. PMID- 7474409 TI - Intensive chemotherapy with cisplatin, doxorubicin, cyclophosphamide, etoposide and granulocyte colony-stimulating factor for advanced thymoma or thymic cancer: preliminary results. AB - A study was conducted to evaluate the impact of cisplatin, doxorubicin, cyclophosphamide and etoposide (PACE) with granulocyte colony-stimulating factor (G-CSF) on advanced thymoma or thymic cancer. Between August 1989 and December 1994, 14 patients with invasive, metastatic or recurrent thymoma or thymic cancer were treated with cisplatin (80 mg/m2, on day 1), doxorubicin (45 mg/m2, on day 1), cyclophosphamide (800 mg/m2, on day 1) and etoposide (80 mg/m2, on day 1-3) with G-CSF (90 micrograms/m2, on day 5-18) at the National Cancer Center Hospital, Tokyo. Courses were repeated every 3 or 4 weeks for a maximum of 4 cycles. Twelve patients were treated with 2 or more courses of PACE. Two patients were treated with only one course, one refused and another required emergency thoracic radiotherapy after one course of PACE. Six patients had partial responses (3 thymomas and 3 thymic cancers) but there were no complete remissions (response rates, 42.9%; 95% confidence interval, 17.7% to 71.1%). Moderate hematological toxicities were observed: grade 3 or 4 leukopenia, neutropenia, anemia and thrombocytopenia in 10, 13, 8 and 6 patients, respectively. Six patients developed infections that required antibiotics. Surgical resection or thoracic radiotherapy after PACE treatment was performed in 2 and 7 patients, respectively. The overall median survival time was 14.7 months (range, 5.9 to 59.7 months). For 9 patients who had received no prior treatment before chemotherapy, the median survival time was 8.9 months, and one patient survived for 4 years and is still alive. In conclusion, PACE with G-CSF frequently produces objective remissions in patients with advanced thymoma or thymic cancer. A large-scale intergroup study is necessary to determine the impact of this regimen on advanced thymoma and thymic cancer. PMID- 7474411 TI - Pneumocystis carinii pneumonia during treatment for recurrent breast cancer: a case report. AB - We describe a case of Pneumocystis carinii pneumonia (PCP) in a woman with recurrent breast cancer without human immunodeficiency virus (HIV) infection. The PCP was associated with severe lymphocytopenia due to treatment with anticancer agents in combination with granulocyte colony-stimulating factor (G-CSF). Despite the severe lymphocytopenia, the total leucocyte count never fell below 3000/mm3 during the treatment. It was difficult to determine whether the patient's respiratory failure was caused by severe infectious pneumonia, hypersensitivity pneumonia or pneumonitis carcinomatosis. She was treated with steroid for suspected drug-induced hypersensitivity pneumonia. However, as her condition did not improve, PCP was suspected, and sulfamethoxazole-trimethoprim was administered. At the same time, anticancer drugs were administered to half the progression of the cancer, since lymphangitis carcinomatosa was also suspected. The severe respiratory failure did not improve, and the patient died on day 23 after admission. At autopsy, the cause of death was confirmed to respiratory failure due to PCP. PMID- 7474410 TI - Patient with extensive small cell lung cancer given moderately dose-intensified chemotherapy and peripheral blood stem cell support. AB - Although recent progress in multi-drug chemotherapy has increased the median survival of patients with extensive Small cell lung cancer (SCLC) to 7 months, 2 year survivors are still exceptional. We describe a 40-year-old man with extensive SCLC involving the lungs and systemic nodes who was initially treated with 6 cycles of an alternating combination regimen consisting of cyclophosphamide, doxorubicin, vincristine, cisplatin and etoposide. After the second cycle of the regimen, Peripheral blood stem cells (PBSC) were harvested and frozen. The patient then received a moderately dose-intensified regimen consisting of cisplatin (80 mg/m2) and etoposide (500 mg/m2 X 3), followed by infusion of the thawed PBSC. Hematopoietic recovery was rapid, and the patient remained tumor free for 12 months until multiple tumor recurrence. PMID- 7474412 TI - Report of the Eighth International Symposium of the Foundation for Promotion of Cancer Research: basic and clinical research in colorectal cancer. PMID- 7474413 TI - [Indication of bone marrow transplantation]. PMID- 7474414 TI - [Tests for donors of blood and bone marrow]. PMID- 7474415 TI - [Clinical and histologic diagnosis of graft-versus-host disease]. PMID- 7474416 TI - [Detection of hematopoietic stem cells using automated blood cell counter and flow cytometry]. PMID- 7474417 TI - [Hemopoietic colony formation in semisolid and liquid culture system]. PMID- 7474418 TI - [Splenic colony forming method]. PMID- 7474419 TI - [Gene diagnosis for patients with leukemia]. PMID- 7474420 TI - [Laboratory examinations and therapy for infections after bone marrow transplantation]. PMID- 7474421 TI - [Allogenic bone marrow transplantation]. PMID- 7474422 TI - [Blood level monitoring of immunosuppressive agent]. PMID- 7474423 TI - [Collection of bone marrow fluid]. PMID- 7474424 TI - [Mobilization and harvest of peripheral blood stem cells]. PMID- 7474425 TI - [Preservation of hematopoietic stem cells]. PMID- 7474426 TI - [Ex vivo purging for autologous hemopoietic progenitor cell transplantation]. PMID- 7474427 TI - [Utilization of bone marrow donor bank]. PMID- 7474428 TI - [Function of the bio-clean room for bone marrow transplantation]. PMID- 7474429 TI - [Serological typing of HLA]. PMID- 7474430 TI - [Autologous bone marrow transplantation]. PMID- 7474431 TI - [DNA typing of HLA]. PMID- 7474432 TI - [Serum lipoprotein and atherogenicity--relationship with particle size and composition of low density lipoproteins, and their modifications by oxidation and glycation]. AB - Low density lipoprotein (LDL) has been recognized commonly as an atherogenic lipoprotein. Especially, small dense LDL is considered to be more atherogenic than large buoyant LDL. It is contained in polydisperse LDL with various sizes and densities. Although the causative mechanism of LDL heterogeneity has not been elucidated clearly yet, it is supposed to be derived from the heterogeneous mechanism of metabolic processing of the VLDL(very low density lipoprotein)-LDL cascade different from the common pathway. It might be due to quantitative and qualitative abnormalities of secreted VLDL, different susceptibilities to lipoprotein lipase as well as hepatic lipase, and the change of receptor binding abilities of LDL. The development of polydisperse LDL has a genetic background. However, it must be influenced by some environmental factors, since the heterogeneity of LDL is restored completely to a normal monodisperse pattern concomitantly with the decreased serum triglyceride level by dietary caloric restriction. LDL is well known to be oxidized or glycated spontaneously in vivo and in vitro. Such modified LDL loses the binding affinity to the classical LDL receptor and is taken up by monocyte-derived macrophages through scavenger receptors. Since the scavenger receptor system is not down regulated, it promotes the formation of foam cells in the arterial wall accumulating cholesterol and other lipids by excess LDL uptake. Small dense LDL is considered to be easily oxidized or glycated compared with large buoyant LDL. Therefore, it might increase further the atherogenic potency by such modifications. PMID- 7474433 TI - [Gene technology to overproduce the enzymes useful as diagnostic reagents]. AB - We cloned the genes encoding the microbial enzymes described below and discussed the possibility as the diagnostic reagents: Creatinase(CR) and sarcosine oxidase(SOX) are useful for a diagnostic measurement of creatinine in combination with creatinase. Bacillus sp. B-0618 produces both CR and SOX enzymes when grown in the presence of an inducer, choline chloride. We cloned these two genes encoding CR and SOX by using recombinant DNA techniques. We found that the SOX encoding gene is located upstream from the CR-encoding gene. When the CR- and SOX encoding genes were independently inserted into the pUC-based vector and introduced into Escherichia coli, the transformants produced 15.5-fold more CR and 50-fold more SOX than Bacillus sp. B-0618, respectively, in the absence of the inducer. Although the Bacillus SOX is a flavoprotein, we created an FAD-free SOX by site-directed DNA mutagenesis. The mutant protein no longer expressed the SOX activity. Cholesterol esterase is useful for a diagnostic measurement of total cholesterol in combination with cholesterol oxidase. We cloned a gene encoding the cholesterol esterase enzyme. When the gene was introduced into the non-producing Streptomyces strain, the transformant produced the cholesterol esterase. Phospholipase A2 is a diverse family of enzymes that hydrolyze the sn-2 fatty acyl ester bond of phosphoglycerides producing free fatty acids and lysophospholipids. The enzymes are present in pancreatic juice and in the venoms of snakes and bees, where they serve digestive functions.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7474434 TI - [A PCR method for examining diarrhea--Escherichia coli]. AB - To identify diarrhea--Escherichia coli(E. coli), especially Enteroinvasive E. coli(EIEC), Verocytotoxin-producing E. coli(VTEC), and Enterotoxigenic E. coli(ETEC), confirmation of pathogenic factors is necessary. We developed a system using the polymerase chain reaction(PCR) that simultaneously confirms the pathogenic genes of these E. coli, and have used it in routine examination. This method is simple and does not require examination for each pathogenic factor. Multiple pathogenic genes could be rapidly detected. PMID- 7474435 TI - [Thalassemia]. AB - Gene analysis of thalassemia started more than 10 years ago. The analysis has been considerably facilitated by the advent of PCR. There are over 20 kinds of mutations in Japanese beta-thalassemia known to date, among which 8 mutations comprise 75% of all families with beta-thalassemia. Several simplified methods are available for these "common" mutations. Here, are introduced two simple methods; amplification refractory mutation system(ARMS) and gap PCR for analyses of beta-thalassemia(one-point mutation) and alpha-thalassemia(large DNA deletion), respectively. These methods are easily applicable to routine DNA analysis. PMID- 7474436 TI - [Diabetogenes; which cause type II diabetes mellitus]. AB - Glucose homeostasis depends upon a balance between glucose production by the liver and glucose utilization by insulin-dependent tissues, such as muscle and fat. Insulin, secreted by the pancreatic B cell, inhibits hepatic glucose output and facilitates glucose utilization in the muscle and fat tissues. In the diabetic patients, there are three types of pathological defects; (1) inability of the B cell to secrete normal insulin in an accurate fashion, (2) increase in the hepatic glucose output, and (3) decrease in glucose metabolism at the muscle and fat tissues. Candidate genes which cause the first defect are the insulin gene, the glucokinase gene and the mitochondrial tRNA gene. The glucokinase gene causes the second defect, too. Those which cause the third defect are the insulin receptor gene and glycogen synthase gene. However, these genes in total account for 3% or less of type II diabetes. Further analyses of systems of both insulin secretion and insulin receptor-signaling at a molecular level will help elucidate the other diabetes candidate genes. PMID- 7474437 TI - [Determination of gene mutation of silent serum cholinesterase and its epidemiologic characters in the Japanese]. AB - We detected 121 individuals with silent type of serum cholinesterase from 36 families in Japan. DNA analysis totaling 37 members of eleven blood unrelated families were carried out by four useful methods, namely, 1) PCR-SSCP analysis, 2) dot blot hybridization (DBH) with the use of synthetic oligonucleotide probe, 3) restriction endonuclease analysis (REA) and 4) direct sequencing analysis. Their mutations were classified into four groups, namely, 1) a G-->C transversion at codon 365, 2) a frameshift mutation with insertion of an extra A at codon 315, 3) an A-->G transition at codon 128 and 4) a C-->A transition at codon 400. The three procedures including (PCR-SSCP, DBH, REA) without the use of radio labeled materials (non-RI) are recommendable for the analyses. However, the direct sequencing analysis of bases with RI might be, at present, necessary for the final identification. PMID- 7474438 TI - [Application of molecular diagnosis to human lung cancer]. AB - Recent developments in the molecular genetics of human cancers shows the importance of multiple genetic alterations in the pathogenesis of these lesions. DNA diagnostic techniques are being introduced rapidly into the clinical laboratory setting. 1) In lung cancer, several oncogenes and tumor suppressor genes, such as ras, myc, p53, RB, allelic loss of chromosomes, play very important roles. These genetic changes are being applied to cancer diagnosis, prediction of prognosis or disease metastasis, or response to treatment. 2) Drug resistance is one of the major problems of current lung cancer chemotherapy. Identification of the molecular marker for drug resistance, like DNA topoisomerase gene mutation, in clinical samples will be of great help for choosing chemotherapy regimens. 3) Interindividual differences in susceptibility to lung cancer may be screened using genotyping of the P450IA1 and GSTmu genes. To develop newer diagnostic and therapeutic approaches, detailed investigation of the molecular pathogenesis of lung cancer using clinical samples is essential. I review the present status on these applications of genetic markers to lung cancer diagnosis in this article. PMID- 7474439 TI - [Gene diagnostics of gender verification test in competitive sports]. AB - The gender verification test (GV) was carried out by gene diagnostics at the 12th Asian Games in Hiroshima, 1994. Human genomic DNA samples were extracted from the hair root of female competitors. Polymerase chain reaction (PCR) was used to prove the absence of the sex determining region of Y-chromosome (SRY) in order to confirm the femininity, and detection of pseudo autosomal boundary region of Y chromosome (PABY) in order to confirm that the DNA is extracted from the hair root and the femininity can again be confirmed. A total of 1,300 female competitors were screened. We conclude that this testing is useful for the GV, because hair sampling is easy, the method is simple and there is little contamination. PMID- 7474440 TI - [Quality control of fine needle aspiration cytology of breast lesions]. AB - From 1988 to 1993 we treated 1,310 cases of fine needle aspiration cytology of breast lesions. The results were positive for malignancy in 218, suspicious in 134, negative in 857, and inadequate specimen in 101. Histological examinations were performed in 461 cases; 243 malignancies and 218 benign lesions. The cytological diagnosis of these cases was positive in 211, suspicious in 91, negative in 142 and inadequate specimen in 17. The cytologic diagnosis of the cases of histological malignancy was positive in 198, suspicious in 24, negative in 17 and inadequate specimen in 4. Therefore we have 17 false negative cases. The histological diagnoses in these cases were scirrhous carcinoma (7), papillotubular carcinoma (6), invasive lobular carcinoma (2), mucinous carcinoma (1) and angiosarcoma (1). There were 13 cases of cytologically false positive cases; the histological diagnoses were fibroadenoma (4), mastopathy (4), papilloma (3), adenosis (1) and xanthogranuloma (1). For over six years, we have used a computer system incorporating both cytologic and histologic records. This system is very useful in quickly review the correlations of cytologic-histologic diagnoses. In addition to this system, a close relationship between clinical to pathological department is necessary in the quality control of diagnostic cytology. PMID- 7474441 TI - [Exercise test in respiratory system]. AB - A ramp exercise test was thought to be adequate especially for evaluation of patients with respiratory system diseases. First, we told about the significances, the methods and apparatus, the arrangements for workload and the matters that demand special attentions of this exercise test. Next, various indices which were obtained during ramp exercise and the estimations of them were explained. Furthermore, we referred to the exercise-limiting factors for patients with COPD and pulmonary fibrosis as symmetrical respiratory system diseases. PMID- 7474442 TI - [Exercise testing in cardiac patients]. AB - Symptom-limited incremental exercise tests have been used, to estimate the severity of cardiovascular disease and the patients' daily activity. However, there is a considerable amount of interest in obtaining submaximal measurements of aerobic function rather than parameters requiring maximal exercise effort. We compared the parameters obtained during the incremental exercise with those during the 6 minutes of moderate constant work rate exercise. Peak oxygen uptake (VO2) and the anaerobic threshold were significantly decreased in patients with cardiovascular disease as compared to normal subjects. The slope of the increase in carbon dioxide output (VCO2) to the increase in VO2 (delta VCO2/delta VO2) above the anaerobic threshold was significantly increased and the slope of the increase in VO2 to the increase in work rate (delta VO2/delta WR) was significantly decreased in patients with cardiovascular disease. The anaerobic threshold was found to occur at the work rate above which left ventricular function decreased during exercise in these patients. The time constant of VO2 during and following recovery from 6 minutes of 50 watts of constant work rate exercise was significantly longer (the kinetics of VO2 were slower) in patients with cardiovascular disease than in normal subjects. The time constant of VO2 was significantly negatively correlated with peak VO2 and maximum work rate obtained during the incremental exercise.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7474443 TI - [Changes in coagulation parameters during the clinical courses of recipients of living-related partial liver transplantation]. AB - We had rare opportunities to examine changes in fibrin degradation products (FDP) D-dimer (DD), thrombin-antithrombin III complex (TAT), plasmin-alpha 2-plasmin inhibitor complex (PIC) and other coagulation parameters during the clinical courses of living-related partial liver transplantation (LRPLT). In seven out of eight recipients without severe rejection and/or disseminated intravascular coagulation (DIC), FDP-DD values reached their maximum at 5 to 10 days after transplantation, then gradually decreased. On the other hand, TAT values rose to the maximum at anhepatic or reperfusion phase of liver transplantation. These data represent hypercoagulation in consequence of tissue thromboplastin activation after extensive operation. Changes in PIC, tissue-type plasminogen activator, and plasminogen activator inhibitor-1 (PAI-1) in the clinical course of case 1 suggested that fibrinolysis was suppressed by relatively elevated level of PAI-1 around the operation, but thereafter was adversely accelerated by relatively lower levels of PAI-1. In comparison with patients with DIC, TAT was much higher but PIC was significantly lower in recipients of LRPLT. These findings indicated that marked hypercoagulation and mild to moderate hyperfibrinolysis occurred in recipients of LRPLT. PMID- 7474444 TI - [Detection of hepatitis B virus by using polymerase chain reaction and nonradioactive DNA probes. I. Identification of mutations in the precore region by PCR-direct sequencing and ASO probe method]. AB - Seroconversion from hepatitis B e antigen (HBeAg) to anti-HBe antibody (anti-HBe) frequently occurs in hosts chronically infected with hepatitis B virus (HBV). Further, this phenomenon is related to a point mutation from guanine to adenine at nucleotide 83 in the precore region of HBV, which, converts codon 28 for tryptophan (TGG:W) to a translational, stop codon (TAG:X). Therefore, we decided to examine HBV in sera from patients for mutations in the precore region by a simple allele-specific oligonucleotide (ASO) probe method. Direct sequencing was first performed on DNA fragments amplified by the polymerase chain reaction in order to establish whether there were mutations in the precore region. Subsequently, specific DNA probes were applied to detection of mutations in the precore region. Subsequently, specific DNA probes were applied to detection of mutations in the precore gene. Five unknown mutations (I10N, C12W, C14S, V17F and A19D), three known mutations (I9V, W28X and G29D) and for novel nucleotide insertions were identified in anti-HBe positive sera. By using seven nonradioactive probes, we could determine the mutations at codons 9, 28 and 29 in anti-HBe positive sera. The W28X mutation was found in anti-HBe positive but not in any of HBeAg positive sera. Meanwhile, wild-type strains of HBV were detectable in sera from patients who were positive to HBeAg or anti-HBe. This ASO probe assay could determine in a few days the mutations in the precore region of HBV, especially including the defect to prohibit the synthesis and secretion of HBeAg. PMID- 7474445 TI - [Evaluation of the polymerase chain reaction for diagnosis of tuberculous meningitis]. AB - Since Shankar et al used polymerase chain reaction (PCR) for detection of Mycobacterium tuberculosis (M. tuberculosis) in cerebrospinal fluid (CSF), there have been numerous reports on PCR assay for diagnosis of tuberculosis meningitis (TBM). The PCR assay have been recognized to be a rapid method for diagnosis of TBM, however there are problems of PCR sensitivity when dealing with CSF samples containing small amount of M. tuberculosis DNA. Comparing direct PCR with nested PCR, we studied on the evaluation of PCR for diagnosis of TBM. In present study the nested PCR were positive in all CSF specimens from 4 patients with TBM, but we could not detect M. tuberculosis DNA by only the direct PCR. Because nested PCR amplification improved the sensitivity and specificity, the nested PCR assay will be necessary for rapid diagnosis of TBM on the clinical laboratory. PMID- 7474446 TI - [MALT lymphoma of large intestine as multiple large polypoid lesions]. AB - We report a case of mucosa associated lymphoid tissue (MALT) lymphoma in the large intestine in a 38-year-old Japanese female. She developed a dull pain in the right lower abdomen and was found to have ileocecal intussusception. The terminal ileum, cecum and ascending colon were resected. Macroscopically, multiple polypoid lesions were found. Although some authors reported that MALT lymphoma of the colon tend to be solitary, the present case showed seven lesions. Two of the polypoid lesions in the present case were marked large. No such large polypoid MALT lymphoma has been described to our knowledge. A histological and immunohistochemical study revealed those seven lesions to be low grade B cell lymphomas of MALT type. PMID- 7474447 TI - [Detection of the exoskeleton of Sarcoptes scabiei in histological sections using Fungiflora Y]. AB - Fungiflora Y was applied to a deparaffinized section of a biopsied skin with scabies, a section of a brain infected with Cysticercus cellulosae, a section of a lung infected with Pneumocystis carinii and a section of a cockroach. The exoskeleton of the mite and the cockroach were stained with Fungiflora Y and were visualized as a greenish-blue fluorescence under incident illumination with ultraviolet light, whereas the Cysticercus cellulosae and the Pneumocystis carinii were not visualized at all. Fungiflora Y, which has been reported to stain fungi, is likely to detect arthropods exoskeleton in histological sections. PMID- 7474448 TI - [Ictal electroencephalographic recordings of patients with seizure]. AB - We analyzed the ictal electroencephalographies (EEGs) in 75 seizures of 73 patients. Eighty percent of the patients were below the age of 20. The seizures consisted of 35 generalized seizures, 24 partial seizures and 16 pseudoseizures. The ictal EEG of true seizures showed changes with a spike in 23 cases, and change without spike (eg. rhythmic activity, desynchronization) in 32 cases. Four ictal records could not be evaluated due to artifacts. Because half of the patients with pseudoseizures also had epilepsy, their ictal EEG examinations were very useful. The ictal EEG examination could be performed successfully on 96% of the patients with a seizure frequency of more than six a day, 52% of them with one to five seizures a day and eleven percent of them with less than one seizure a day. Seventy-five percent of ictal EEG recordings were performed when the patient was awake. The cassette of electrodes, originally made in our department, was very useful in recording the ictal EEG of children when they were awake. PMID- 7474449 TI - [Laboratory evaluation of autonomic function]. AB - The sympathetic and parasympathetic systems both contain efferent and afferent neurons. The efferent pathway consists of preganglionic neurons with myelinated axons the cell bodies of which are within the central nervous system and postganglionic neurons with unmyelinated axons the cell bodies of which are peripheral. The preganglionic sympathetic fibers arise in the intermediolateral column of the thoracic and upper lumber cord segments, and reach the sympathetic chain whence postganglionic fibers are distributed to the whole body, where they have particular influence on the control of blood pressure, heart rate, swating and pupillary responses. The parasympathetic outflow is grouped into both brainstem nuclear complex and spinal sacral segments. The activities of the sympathetic and parasympathetic systems are partially reflex in nature, which, however, are controlled by numerous descending influences integrated in the hypothalamus and reticular formation. Orthostatic hypotension is the most dramatic circulatory expression of autonomic failure. As one stands, the pull of gravity pools blood in distensible veins below the heart. Diminished venous return reduces cardiac output and unless autonomic compensatory mechanism baroreflex function performs, subsequent blood pressure fall occurs. This hemodynamic responses to upright posture can be evaluated by the head-up tilt test. More than a 30-mmHg fall of the systolic blood pressure is thought to be an abnormal response; orthostatic hypotension. The next test to study the orthostatic hypotension is to evaluate the site of the baroreflex arc lesions. Plasma noradrenaline (NA) and vasopressin (AVP) responses to the head-up tilt test are available to detect the site of the lesion.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7474450 TI - [Clinical investigation for polymyositis and related disorders]. AB - The clinical investigation for inflammatory myopathies, which include polymyositis (PM), dermatomyositis (DM) and others, was outlined. The serum creatine kinase (CK) activity increases in the majority of cases of inflammatory myopathies. However, the cases of myositis associated with connective tissue diseases tend to show normal or moderately elevated CK activity. Among the isoenzymes of CK, the MB fraction can increase in the course of treatment as it can originate from regenerating muscle fibres. The macro CK type 1 was reported to appear in association with myositides. Varieties of autoantibodies in the serum such as Jo-1 and Ku have been studied. The Jo-1 antibody is frequently detected in the cases of PM associated with interstitial pulmonary fibrosis. Examination of the heart and lungs is necessary, and so is a search for malignant neoplasms in the cases of DM. Muscle biopsy is mandatory for diagnosing PM, DM and other inflammatory myopathies. Among the latter, inclusion body myositis and granulomatous myopathy need to be identified before treatment as they generally respond poorly. Histological changes of inflammatory myopathies are often distributed unevenly. The magnetic resonance image and ultrasonography are helpful in estimating the distribution of the lesion and therefore in deciding the site of biopsy. Ultrastructural observation of the muscle showed invasion of activated lymphocytes under the basement membrane of the muscle fibres causing degeneration of the myofibrils. The subset analyses of infiltrating cells revealed considerable alterations after the steroid pulse therapy. PMID- 7474451 TI - [Neuroimaging and pathology of the spinal cord in compressive cervical myelopathy]. AB - Magnetic resonance imaging (MRI) has enabled us to see the spinal intramedullary pathology as differences in signal intensity. Intramedullary high intensity lesions were observed on T2-weighted MRI in patients with cervical spondylotic myelopathy (20.0%) and ossification of the posterior longitudinal ligament (OPLL) of the cervical spine (25.7%). The frequency of this findings was proportional to the clinical severity of myelopathy and degree of spinal cord compression. The pathophysiological basis of such signal abnormality was presumed to vary from acute edema to chronic myelomalacia. The intramedullary lesion on MRI is considered to be the main site of lesion responsible for the neurological symptom because of a good correlation between the neurological level and high intensity level. We found from nine autopsy cases of OPLL that there are distinct differences in severity and extent of pathological changes between the spinal cord with a boomerang-shaped cross-section and that with a triangular-shaped cross-section. In the boomerang-shaped cases, major pathological changes were restricted to the gray matter and the white matter was relatively well preserved. Secondary wallerian degeneration was restricted to the fasciclus cuneatus the fibers of which were derived from the affected segments. In the cases of a triangular shape, pathological changes were more severe, both white and gray matter were involved. There were severe pathological changes over more than one segment, and both descending degeneration of the lateral pyramidal tracts and ascending degeneration of the posterior column, including the fasciclus gracilis, were observed. In conclusion, it is clinically very important to understand the pathological basis of the compressed spinal cord on neuroimages. PMID- 7474452 TI - [Biochemical studies on AIDS dementia complex--possible contribution of quinolinic acid during brain damage]. AB - AIDS dementia complex (ADC) is a complex, progressive neuropsychiatric syndrome seen in 60-70% of the patients with AIDS. The structural and functional changes associated with ADC may be the result of a variety of indirect mechanisms mediated via activated brain cells or/and virus that produce neurotoxins including N-methyl-D-aspartate receptor agonist (eg, quinolinic acid, glutamate), cytokines, gp 120 and nitric oxide. The level of the neurotoxin and kynurenine pathway metabolite, quinolinic acid, is increased in the brain and CSF of HIV-1 infected patients, and is correlated with quantitative measures of neurologic impairment. Importantly, increased CSF and brain levels of QUIN also occur in other inflammatory neurologic diseases (bacterial, viral, fungal and parasitic infections, meningitis, autoimmune diseases and septicemia), independent of HIV-1 infection. Therefore, QUIN and other neuroactive kynurenine pathway metabolites may be final common mediators of neurologic dysfunction in a broad spectrum of inflammatory neurologic diseases. Conversion of L-tryptophan to QUIN has also been demonstrated in vitro in both brain tissue following macrophage infiltration, and in macrophages stimulated by interferon-gamma or HIV infection. Macrophages in vitro have a high capacity to synthesize QUIN following exposure to interferon-gamma, tumor necrosis factor-alpha, IL-1 beta and IL-6, compared to cells derived from other tissues. Notably, the concentrations achieved in the macrophage incubates exceeded the levels found in the CNS of HIV-1-infected patients, and exceeded the concentrations shown to be neurotoxic in vitro. We hypothesize that increased kynurenine pathway metabolism following inflammation reflects the presence of macrophages and other reactive cell populations at the site of brain infection. Strategies to attenuate the neurotoxic effects of kynurenines, such as inhibitors of kynurenine pathway metabolism and cytokine antibodies may offer new approaches to therapy. PMID- 7474453 TI - [Pathophysiology on the disorder of neuromuscular transmission defects]. AB - Neuromuscular transmission defects are often puzzling but challenging diseases for the pediatric and adult neurologists. Some of these disorders entail severe or even life-threatening disability: Most are treatable, but effective therapy requires precise diagnosis sometimes difficult to make especially in congenital defects. The diagnosis rests on the combination of clinical data, the electromyogram and additional studies that may include microelectrode analysis of neuromuscular transmission, ultrastructural and cytochemical studies of neuromuscular junction (NMJ) and biochemical/molecular genetic studies on muscle specimens. Understandably, these studies often depend on the collaboration of several investigators. In each myasthenic disorder, an abnormality affects neuromuscular transmission directly or causes secondary derangements that eventually affects transmission. Namely, lesions of both the presynaptic and/or postsynaptic area manifest a variety of clinical findings. Here was discussed the pathophysiology on myasthenia gravis and Lambert-Eaton myasthenic syndrome of abnormal autoimmunity, congenital myasthenic syndromes of a chromosomal defect, and poisoning of organophosphorus, botulism and some other toxins. PMID- 7474455 TI - [Morphologic analysis of blood cells]. AB - Accompanying hematology laboratory automation, the frequency of morphological observation of blood cells tends to decrease. Although 5-part differentials are performed by automated systems, counting of abnormal cells such as immature cells or leukemia cells still must be performed manually using smears. In the automated hematology laboratories, technicians more frequently find abnormal cells than before. Therefore, poor microscopic observation of blood cells markedly affects the diagnosis. Continuous training of the technologist is mandatory. Also, preparation of smear samples is important, i.e, well prepared slides must be stained properly with an appropriate dye. Technicians must be familiar with staining technology together with the knowledge of the characteristics of several staining methods. On the other hand, the technician's ability to differentiate white blood cells depends on the instructor's knowledge. This makes quality control and the standardization of differentials difficult. Still the problems must be solved under the cooperation of the various societies concerned. PMID- 7474454 TI - [Present and future of classification for white cells and leukemic cells in the routine clinical laboratory]. AB - Renovation of laboratory medicine has been greatly advanced in the past decade. White cell differentiation has been done with the automated blood cell counter with high reliability with the combination of flow cytometry. Specific monoclonal antibodies have made FAB classification more precise, while the traditional cell classification still gives some extent of uncertainty for the decision of cell typing. High resolution chromosome analysis and DNA analysis enable one to define genetical leukemia types and patients' prognosis. Clinically the hematological data reported from the routine laboratory are regarded as very reliable, but an interpretation should be made carefully to select the patient treatment and prediction of prognosis. Here, four speakers were selected to show the present problems and some resolution for identification of white cells and leukemia cells. The importance of standardization work was also discussed for identification of white cells and leukemia cells examined by the traditional Romanovsky stain, automated blood cell counter, flow cytometric method, and chromosomal and DNA analytic methods. We concluded that all those methods should be standardized and interpretation would be harmonized to give common answers for clinical decisions. PMID- 7474456 TI - [Further evolution and leukocyte differential using an automated blood cell counter]. AB - In the leukocyte differential, morphological differential has been usually performed with chromatoaffinity of blood cells, but physical differential is recently performed with biophysicochemical characteristics of blood cells using an automated blood cell counter. The trend of the leukocyte differential has changed to physical differential (automated count method) from morphological differential (eye count method). In the automated count method, leukocytes are differentiated into neutrophils, lymphocytes, monocytes, eosinophils and basophils according to the predetermined region of each leukocyte population on the cytogram using an automated blood cell counter. An automated count method is very useful for the screening test in the laboratory because of its high precision and accuracy for the normal samples. However, it is not easy to identify abnormal cells such as leukemia cells by an automated count method. Therefore, an automated count method dealing with various suspect flags generated using an automated blood cell counter must be used for such abnormal samples. In case of absolutely few leukemia cells in the peripheral blood during complete remission, the automated count method can not detect a leukemia cell and no suspect WBC flags are recognized by the automated blood cell counter. Development of an automated blood cell counter with a higher efficiency than now and a standard automated count method is awaited in the clinical laboratory. PMID- 7474457 TI - [Immunophenotypic analysis of lymphocyte subpopulation and hematological malignancies]. AB - Determination of the human lymphocyte subpopulation selected by immunofluorescence and the phenotypic analysis of hematological malignant cells by laser flow cytometry have become popular and useful tests in various laboratories. However, several lines of evidence have questioned the accuracy and reproducibility of these analysis. We examined the problems of laser flow cytometric analysis to measure the lymphocyte subpopulation and determine the phenotypic expression of hematological malignancies. In lymphocyte subset analysis, no survey has been applied to reveal the accuracy and reproducibility of these tests. We compared the accuracy of gating events and the ratio of lymphocytes using leuco GATE/simul SET analysis to those by manual gate method analysis. We found that there were some patients with SLE in which the accurate lymphocyte subpopulation was difficult to calculate due to the gating of lymphocytes by either method. Furthermore, apparent differences in the lymphocyte population were observed between these methods. In the phenotypic analysis of hematological malignancies, there have been several problems over 30% of the total cells had to be abnormal cells. Second, the malignant cells were difficult to gate unless the information of the size, shape and cellular density were obvious. Third, the phenotype of malignant cells were often different from that of the normal matured cells in the some lineage. However, flow cytometric analysis was useful to determine the cell lineage of peroxidase-negative cells and to diagnose the hybrid leukemia. In summary, the phenotypic analysis using flow cytometry and various monoclonal antibodies are clinically useful tests to diagnose the immunological disorders and hematological malignancies. However, there remain several problems to be solved in the near future. PMID- 7474458 TI - [Detection of Mycobacterium tuberculosis by amplification of mycobacterial DNA from pleural fluid and gastric juice: the comparison of IS6110 gene with Amplicor mycobacterium]. AB - The polymerase chain reaction (PCR) using oligonucleotides based on the repetitive sequence (IS6110) of Mycobacterium tuberculosis and Amplicor Mycobacterium, which combines a PCR with the hybridization assay, were evaluated for detection of M. tuberculosis in pleural fluid and gastric juice. The detection limits of these two assay systems for cultured M. tuberculosis were less than 2 cells/ml, as compared with 200 cells/ml by culture. A total of 42 pleural fluid and 94 gastric juice specimens were examined. A total of 5 pleural fluid and 5 gastric juice were culture positive for M. tuberculosis. Only the PCR gave positive results in 2 pleural fluid of which laboratory findings are characteristic of tuberculous pleuritis, and in one gastric juice of the patient who was diagnosed as having pulmonary tuberculosis. Sensitivity, specificity and positive predictive value for IS6110 gene were 75%, 94% and 71.1%, respectively, in pleural fluid. Two of three positive specimens for IS6110 gene from pleural fluid were negative for Amplicor Mycobacterium. These specimens resulted in positive with Amplicor Mycobacterium when solvent extracted DNA was used. Both the PCR systems had the same sensitivity (80%), specificity (98.8%) and positive predictive value (83.3%) in gastric juice. The PCR systems would be useful for rapid detection of M. tuberculosis without long term culture and for detection of non-cultured one from pleural fluid as well as gastric juice. However, when the presence of an inhibitor of PCR is suspected in a specimen, DNA should be purified from the specimen before amplification. PMID- 7474459 TI - [Changes in plasma tissue plasminogen activator (t-PA) and plasminogen activator inhibitor 1 (PAI-1) by the interferon treatment for chronic hepatitis C]. AB - To determine the effects of interferon (IFN) treatment for chronic hepatitis C on vascular endothelium, we measured the concentrations of tissue plasminogen activator (t-PA) and its inhibitor (PAI-1) in the plasma from patients before and after IFN treatment for 14 consecutive days. The plasma t-PA and PAI-1 levels were measured before and after treatment. The plasma t-PA level was significantly increased after IFN treatment (p < 0.01) but no significant difference in plasma PAI-1 level was observed before and after treatment. The ratio of t-PA/PAI-1 was significantly increased after IFN treatment (p < 0.05). These changes may be caused by the effect of IFN on endothelium, leading to an activation of the endothelium derived fibrinolysis factors. Increase in plasma t-PA concentration may induce hyperfibrinolysis which may be one of the causes of suborbital hemorrhage. Further study on the fibrinolysis pathway in the blood is necessary to elucidate the mechanisms of the many side effects observed during IFN treatment. PMID- 7474460 TI - [The effects of fitness-type exercise on iron status and hematological status for female college students]. AB - Forty-two female college students underwent fitness-type exercise over a 13 week period. The students exercised an average of two times a week for 30 minute periods at 50% levels of VO2 . max. Iron status and hematological laboratory data were monitored at three points in time; at the initiation of exercise, at the termination of exercise and 3 months later. Serum ferritin levels in a third of the students were lower than 12ng/ml. Red blood cell counts decreased during exercise, and returned to the pre-exercise levels 3 months later. The number of circulatory reticulocytes and mean corpuscular hemoglobin concentration were increased significantly after the exercise and the effects were still observed 3 months later, independent of the iron status. In the lower ferritin group, erythropoietin levels decreased significantly during the post-exercise period. The average serum ferritin levels while showing no significant changes during exercise, increased significantly during the post-exercise period, in the normal ferritin group. We discussed possible mechanisms, among them iron metabolism, by which physical training may facilitate erythropoiesis. However, no definite conclusions have been reached. PMID- 7474461 TI - [A study on immunohistochemical detection of estrogen receptor in the breast cancer tissue--a comparison between routinely paraffin-embedded and frozen sections]. AB - On immunohistochemical detection of estrogen receptor in the breast cancer tissue, we examined whether ER1D5 [Anti-estrogen receptor monoclonal antibody, clone ER1D5(MBL)] using routinely fixed paraffin-embedded section may take the place of ERICA [Estrogen receptor immunocytochemical assay kit (DAINABOT)] using frozen one or not. With 63 breast cancer cases, estrogen receptor was positive in 63.4% of the cases by ERICA, while in 25.4% and 28.6% by ER1D5 in which enhancement of the antigen with microwave heating (high frequency power 500W 5 minutes) and autoclave heating (121 degrees C, 20 minutes) had been carried out. At present, it is unreasonable for ER1D5 to take the place of ERICA. However, it is noteworthy that estrogen receptor is detectable with paraffin-embedded section although the rate of positivity is low. It is desirable that further development of the antibody for estrogen receptor in the breast cancer tissue with paraffin embedded section will be continued. PMID- 7474462 TI - [Excitability of motor cortex with transcranial magnetic double stimulation in the intact man]. AB - To evaluate the excitability of central motor tract, we studied a transcranial magnetic double stimulation with short conditioning-test (C-T) interval of 1-10ms in eight normal volunteers. In addition, H-reflex of the forearm muscle was used to study the effect of the magnetic cortical conditioning stimulus on alpha motoneuron, and the test response evoked by electrical cortical stimulation was also used to examine the effect of the magnetic cortical conditioning stimulus. The subthreshold conditioning and suprathreshold test stimuli were applied, and compound muscle responses were recorded in the relaxed abductor pollicis brevis muscle. There was a decrease of the test response size by the conditioning stimulus at C-T interval of 1-5ms. This attenuation was probably caused by intracortical inhibition. Because the identical magnetic cortical conditioning stimulus produced increase in H-reflex size. Moreover, the test response evoked by electrical cortical stimulus was not suppressed by the magnetic cortical conditioning stimulus; whereas, response evoked by the magnetic cortical test stimulus was suppressed at C-T intervals of 2ms. With the technique of transcranial magnetic double stimulation, therefore, it is possible to evaluate the inhibitory function in the motor cortex. The technique may be of use for pathophysiology, diagnosis and estimation of treatment in the diseases. PMID- 7474463 TI - [Recent advance in surgical treatment for inflammatory bowel disease]. PMID- 7474465 TI - [Role of platelet activating factor on the formation of ischemic colitis]. AB - Role of platelet activating factor (PAF) on the formation of rat ischemic colitis was investigated. High incidence of the formation of ischemic colitis was observed after ligation of marginal vessels of the rat with stricture of the rectum, however, administration of PAF-inhibitor (TCV) decreased incidence of the lesion. Only the lesion in the mucosa occurred after clamping of the marginal vessels of the rat with stricture of the rectum and administration of TCV did not affect the incidence of the lesion. In conclusion, the etiological involvement of PAF on deterioration of the ischemic colitis was suspected. PMID- 7474464 TI - [Influence on glycemic control of improved diabetic gastroparesis by long-term cisapride therapy]. AB - To investigate the effect on glycemic control of improving diabetic gastroparesis, we evaluated symptoms (scored), gastric motor functions (solid and liquid gastric emptying studies and electrogastrography), and glycemic control in 11 patients with diabetic gastroparesis (5 men, 6 women, 50.4 +/- 4.5 years old) before and after treatment with cisapride (15 mg/day p.o., 12 weeks). None of the patients had organic abnormalities on gastrointestinal endoscopy. The dysmotility symptom score (maximum: 18) on cisapride significantly improved from 13.1 to 4.0 (p < 0.01). Retention rates at 15 and 80 minutes after ingestion improved in a solid-food gastric emptying study using a test meal of instant noodles labeled with 37 MBq (1 mCi) technetium-99m (both p < 0.05). Liquid gastric emptying, evaluated using a sulfamethizole technique, also improved but not significantly. Electrogastrography revealed no significant changes after treatment, but the postprandial rate of normal frequency waves tended to increase. Glycemic control was assessed based on HbA1C, fructosamine and M value. There were no significant changes in glycemic control after treatment with cisapride. We conclude that long term administration of cisapride reduced dysmotility symptoms and improved solid and liquid gastric emptying without adversely affecting glycemic control. PMID- 7474466 TI - [Fatty liver and obesity in university students]. AB - Obesity is often associated with coronary heart disease and metabolic disorders. In this study, the relationship between obesity and metabolic disorders and between obesity and fatty liver by ultrasonography was investigated in 307 university students (18-20 years old, men: 196, women; 111). The correlation between Body Mass Index (BMI) and the thickness of subcutaneous fat (ST) was significant between BMI and the ratio between waist and hip circumference (WHR) was more significant in male students (r = 0.838, p < 0.001) than in female students (r = 0.639, p< 0.001). The incidence of fatty liver was significantly higher in male obese students (68.6%) than in female obese students (27.3%). After adjustment for BMI, ST, WHR and sigma glucose, the mean values for serum transaminase, cholinesterase, total cholesterol, uric acid, fasting plasma insulin and sigma insulin were significantly higher in male obese students with fatty liver than in male obese students without fatty liver. The present study suggested that male obese students with fatty liver are more likely to have metabolic disorders than male obese students without fatty liver. PMID- 7474467 TI - [Percutaneous ethanol injection therapy for hepatocellular carcinoma in hepatic dome directly under right diaphragma--the trial of artificial ascites method]. AB - We tried to infuse 40 approximately 100 ml saline solution into abdominal cavity (artificial ascites method) in case of percutaneous ethanol injection therapy (PEIT) for hepatocellular carcinoma (HCC) in hepatic dome directly under right diaphragma (hepatic dome) which is considered as dead space on ultrasonography. Consequently broad visual field was acquired on ultrasonogram and puncture to HCC in hepatic dome was able to be performed very easily. There were no side effects such as difficult hemostasis and peritoneal dissemination. It was concluded that artificial ascites method is very useful to widen visual field of hepatic dome on ultrasonogram and therefore dead space on PEIT disappeared by this method. PMID- 7474468 TI - [Endoscopic lithotomy using sublingual nitroglycerin]. AB - After medical sphincter dilation using sud-lingual nitroglycerin, endoscopic removal of common bile duct stones in 15 patients was carried out. Complete stone removal was obtained in 14 of 15 patients. Only 0.3 mg to 0.6 mg nitroglycerin was needed to give an enough dilation of the orifice for cannulating the basket catheter into the bile duct. Mechanical lithotriptor was smoothly cannulated in one patient because of possible impaction. Blood pressure was dropped transiently in one patient by nitroglycerin, however, the general condition of the patient was stable. This procedure is safe, easy and effective in preserving sphincter function. Acute cholecystitis, a common complication of persistent gallbladder stone after endoscopic sphincterotomy, may be avoided by employing this method. More clinical studies, on long term and in a large scale of patients, are necessary to reduce complications that usually occur after sphincterotomy. PMID- 7474469 TI - [A case of vigorous achalasia, successfully treated with isosorbide dinitrate spray]. PMID- 7474470 TI - [Secondary gastrointestinal amyloidosis: successful treatment with TPN, prednisolone and colchicine therapy. Report of a case]. PMID- 7474471 TI - [A case of primary biliary cirrhosis presenting histopathological similarity to idiopathic portal hypertension with huge splenomegaly]. PMID- 7474472 TI - [A resected case of focal nodular hyperplasia of the liver which could be diagnosed before operation]. PMID- 7474473 TI - [A simultaneous operation for esophageal and cholangiocellular carcinomas--a case of report]. PMID- 7474474 TI - [A case report of the double bile duct associated with malfusion of the pancreatobiliary duct]. PMID- 7474475 TI - [A case of pancreatic arteriovenous malformation--understanding of its pathophysiology with a color Doppler ultrasonography]. PMID- 7474476 TI - [A case report of the asymptomatic islet cell carcinoma]. PMID- 7474477 TI - [Serum complement cold activation is vanished by the interferon therapy for chronic active hepatitis C patients]. PMID- 7474478 TI - [Genetic alteration of gastrointestinal cancers: new tumor marker]. PMID- 7474479 TI - [Clinical evaluation of endoscopic variceal ligation (EVL) using an intensive ligation method]. AB - Endoscopic variceal ligation using an intensive ligation method, or intensive EVL, as we call it, was performed in 17 cases of esophageal varices, from August 1993 to November 1994. Intensive EVL involves making as many ligations as possible from just above the esophago-gastric junction at every session for the complete eradication of varices (F0 and negative Red color sign). Our success rate for the complete eradication of varices was 82.4% (14 out of 17 cases) with the mean number of sessions at 2.2 +/- 0.5 times. The total number of rubber bands used was 20.2 +/- 8.4, with 14.1 +/- 6.0 being used in the initial session alone. The rate of recurrence in complete eradicated cases was 50% (7 out of 14 cases) and the period to recurrence was 6.3 +/- 0.7 months. Although EVL using the intensive ligation method is useful for complete eradication of varices, a high incidence of recurrence in the short term is a concern. We conclude therefore that it is difficult to achieve long term complete eradication of varices with EVL alone, and that additional therapy is needed for long term control. PMID- 7474480 TI - [A study of the relationship between nocturnal intragastric pH and sleep stages of peptic ulcer]. AB - We studied the correlation of the gastric acid secretion and the depth of sleep at night with no medication in 32 cases (19 gastric ulcer, 6 duodenal ulcer patients and 7 healthy controls). Using Memory PH: Monitor intragastric pH values were consecutively measured without antiulcer drugs. The depth of sleep were monitored continuously by recording without any sleeping drugs. In healthy controls, the pH values of awake phase was significantly lower than that of sleep stage 1, 4 and rapid eye movement (REM) phases. The pH values of REM phases were significantly high rather than that of sleep stage 1 and 2 phases. In patients with gastric ulcer, the pH values of awake phases were significantly lower than that of sleep stage 1 and REM phases. The pH values of REM phases were significantly high rather than that of the other sleep stages. On the other hand, in patients with duodenal ulcer, the obvious changes of the pH values were not seen with sleep stages. The pH values of healthy controls and gastric patients were changed with sleep stages. But the pH of duodenal ulcer patients were not changed obviously. Therefore, it is suggested that the dysfunction of autonomous nervous system was influenced the nocturnal intragastric pH in the patients with duodenal ulcer and played one of the important role in the occurrence of duodenal ulcer. PMID- 7474481 TI - [Evaluation of early colorectal cancer with submucosal invasion based on histological architecture]. AB - To clarify the clinicopathological features of tumors with submucosal invasion, especially superficial elevated and sessile ones, histological architectures of 32 cases of submucosal invasive cancers were analyzed. They were classified into 3 types based on cross-section view: PG and NPG of Shimoda's classification and PG'. Histological architectures were drawn in accordance with these findings. In 7 PG-Ca which consisted of PG only, no apparent correlation was found between the tumor sizes and invasion depth of submucosal layer. However, 12 NPG-Ca which consisted of NPG only and 12 Mixed-Ca which include various cross-section views both showed massive invasion into the submucosa with 1 cm or more in tumor size. Therefore these two types were similar in biological behavior in terms of invasion depth. And degree of submucosal invasion tended to increase in the order of PG-Ca, Mixed-Ca and NPG-Ca. Examinating histological architectures of the Mixed-Ca tumors in details, all of these cancers were consisted of both PG and PG'. Of the 12 Mixed-Ca, 91.7% were proved to be PG dominant type. Macroscopically, I s and II a contained 88.9% and 40.0% of Mixed-Ca, respectively. In conclusion, these results suggest that PG' is a subtype of PG, and PG-Ca have a correlation between tumor invasion and alteration from PG to PG' in histological architectures among submucosal cancers. It is important to clarify morphological features of PG' in margin of I s and II a tumors in diagnosing the depth of early colorectal cancers. PMID- 7474482 TI - [Clinical evaluation of a recurrent mode after treatment in patients with small hepatocellular carcinoma]. AB - A total of 96 patients with small hepatocellular carcinoma (HCC) with a maximal diameter of below 2 cm were analyzed according to recurrent factors and recurrent modes after the first treatment. In this study, recurrences were divided into two groups based on the style of recurrences provisionally: intrahepatic metastases (IM, 35 cases) and multicentric recurrences (MC, 22 cases). In patients with IM, the majority of recurrences were observed within 2 years after the first treatment and the primary HCC lesions had strong relation to the degrees of malignancy. On the other hands, there were no association with recurrent intervals and the degrees of malignancy, although the recurrent rate was high in patients with anti-HCV in MC group. The prognosis after the second treatment in MC group was superior to that in IM group (p < 0.05). These results indicate MC group have a good prognosis if early detection and the second accurate treatment are performed. PMID- 7474483 TI - [Transjugular intrahepatic portosystemic shunt for upper gastrointestinal bleeding due to portal hypertension: 2-year results]. AB - We performed transjugular intrahepatic portosystemic shunt (TIPS) in 25 patients with liver cirrhosis who developed upper gastrointestinal bleeding due to portal hypertension. The procedure was successful in 23 patients, in whom portal pressure decreased by 51%. Bleeding was stopped in all 5 emergency for whom other treatments, including sclerotherapy, had failed. During the follow-up period (mean: 367 days), shunt dysfunction due to stenosis was observed in 75% of the patients. However, by percutaneous transluminal angioplasty, the patency could be maintained in 85% of the cases after 1 year and 73% of the cases after 2 years. Re-bleeding was observed in 13% of the cases after both 1 and 2 years. Color Doppler ultrasonography was useful for detecting shunt dysfunction. TIPS seems to be an effective treatment of upper gastrointestinal bleeding due to portal hypertension. PMID- 7474484 TI - [Endoscopic placement of self-expandable metallic stents (Wallstent) for malignant biliary stenosis, with special reference to preventive endoscopic nasal bile drainage against acute obstruction after stenting]. AB - From December 1993 to December 1994, we inserted 14 biliary endoprostheses (Wallstent) endoscopically in 12 consecutive patients (5 men and 7 women; 55-91 years [mean 69.8 years] of age) with malignant biliary stenosis. Successful placement of Wallstent was achieved in all 12 patients. Procedure related death was encountered in one patient (8.3%) who was complicated with emphysematous cholecystitis, combined with a slipping migration of the stent. In 11 of the 12 patients, serum bilirubin levels reduced to less than 2.0 mg/dl. Acute obstruction, which was defined as the obstruction of the stent within 24 hours after the stent placement, occurred in 2 patients (16.7%). Endoscopic nasal bile drainage (ENBD) following stent placement was performed in 4 patients and acute obstruction was not developed in these cases. After endoprosthesis 8 patients were discharged with 6-47 hospital days and the average hospitalized period excluding a case with cerebral infarction was 12.7 days. Excluding one procedure related death, 6 patients died of primary malignancy 39-189 days (average 90.7 days) after endoprosthesis and in 2 cases recurrence of jaundice was observed. The other 4 patients have been alive for 60-174 days (average 102.5 days) without jaundice. The survival rate of the all patients was 64% for 3 months and 32% for 6 months, respectively. In conclusion, we suggested that the endoscopic Wallstent placement would contribute to the improvement of the quality of life for patients with malignant biliary stenosis. However, acute obstruction may be developed in some cases. For the purpose of managing acute obstruction, we recommend ENBD for a few days following the placement of a Wallstent. PMID- 7474485 TI - [Papuloerythroderma associated with gastric cancer; report of a case]. PMID- 7474486 TI - [A case of schwannoma in the transverse colon]. PMID- 7474487 TI - [Interferon induced IBD-like acute colitis--two cases of patients with chronic active hepatitis]. PMID- 7474488 TI - [A case of cholangiocarcinoma with temporary subsegmental fatty liver and intrahepatic portal aneurysm]. PMID- 7474489 TI - [A case of early carcinoma of the gall-bladder arising in adenomyomatosis detected by endoscopic ultrasonography]. PMID- 7474490 TI - [A case of intramural cyst in the gallbladder]. PMID- 7474491 TI - [A case of hemobilia successfully treated with transhepatic embolization]. PMID- 7474492 TI - [A case of occult insulinoma localized by intra-arterial stimulation with calcium and venous sampling technic]. PMID- 7474493 TI - [A case of intraductal papillary adenocarcinoma in adenoma of the pancreas presenting a mutation of Ki-ras codon 12 in the duodenal juice collected during a secretin test]. PMID- 7474494 TI - [Subclinical neuro-psychobehavioral effects in occupational, environmental and community health: methodology and recent findings]. AB - Evoked potentials (short-latency somatosensory, visual and brainstem auditory evoked potentials), event-related potential (P300), electrocardiographic R-R interval variability, peripheral nerve conduction velocity, distribution of nerve conduction velocities, postural balance, neurobehavioral test batteries, and questionnaires and psychiatric interview methods appear to be promising techniques for assessment of subclinical neuro-psychobehavioral effects in occupational, environmental and community health. In this paper, an overview of recent methodology and findings is provided. The available data indicate that these techniques are valid, reliable and standardized, and are easily applied in field studies. PMID- 7474495 TI - [Risk factors for mortality and mortality rate of sumo wrestlers]. AB - We compared the mortality rate of sumo wrestlers with that of the contemporaneous Japanese male population, and inferred the usefulness of an index for predicting longevity in sumo wrestlers. The standardized mortality ratios (SMR) for sumo wrestlers were very high in each period, and also high for ages from 35 to 74. Cox's proportional hazards model analysis revealed that the variables in "nyuumaku" entry year and BMI were statistically significant (p < 0.05) factors in mortality. In the survival curves, the lower BMI group had good life expectancy compared with the higher BMI group. In conclusion, the higher rate of mortality in sumo wrestlers seems to be due to the markedly higher rate of mortality from 35 to 74 years old. In sumo wrestlers, also, this study provides evidence that the higher overweight groups have substantially higher risks for mortality. PMID- 7474496 TI - [Trends of air pollution versus those of consultation rate and mortality rate for bronchial asthma in individuals aged 40 years and above in the Yokkaichi region]. AB - We performed correlation analysis on the relationship between changes in air pollution and the consultation rate for bronchial asthma in the Yokkaichi region, taking effects of various socioeconomic factors into consideration. The effects of changes in air pollution on the mortality rate due to bronchial asthma were also evaluated. 1. Evaluation of annual changes in the simple correlation coefficient between the consultation rate and the concentration of each pollutant showed no significant correlation with a decrease in the air pollutant concentration in the age group less than 10 years old. However, in the middle advanced male and female groups aged 40 years and above, the influence of past air pollution still remained. In addition, the partial correlation coefficients between the consultation rate for bronchial asthma and the degree of pollution, socioeconomic factors, and the rate of heavy smokers were calculated. A significant correlation was observed between the consultation rate for the females in each age group and the rate of patients receiving public assistance. 2. The mortality rate due to bronchial asthma in the polluted area increased rapidly with a time lag of several years after the peak of air pollution but decreased gradually thereafter. Presently, the mortality rate in the polluted area is similar to that in the non-polluted (control) area. 3. The mean age of death due to bronchial asthma was elevated because of a decrease in the deaths of those aged less than 60 years. As a result, the difference in the mean age of death due to bronchial asthma between the polluted area and the control area disappeared. With the recent remarkable alleviation of air pollution, the consultation rate and mortality rate due to bronchial asthma have decreased considerably. However, differences are still observed compared with the control area in some age levels so that continuation of monitoring of air pollution as well as consultation and mortality rates is considered necessary. PMID- 7474497 TI - [Relationship between vibratory sense threshold and blood lead concentration in ceramic color workers and transfer printing manufacturers]. AB - We measured vibratory sense thresholds (VSTs) at 63Hz and 125Hz on the third fingertip of the right hand and on the third toe of the right foot of 74 male workers. The subjects were workers engaged in manufacturing ceramic color and transfer printing paper, whose blood lead (Pb-B) levels were 2-58 micrograms/dl. They were divided into three groups according to the Pb-B levels, namely, below 9, 10-19, and 20 micrograms/dl or more. For statistical analysis, simple and partial correlations, and Scheffe's multiple comparison between the least squares means were used. The VSTs on the fingertip as well as on the toe showed a significant correlation with age. The VSTs at 125Hz on the fingertip were also significantly correlated with alcohol consumption and cigarette smoking. Controlling for age, systolic blood pressure, alcohol consumption and smoking habit, a significant dose-effect relationship was observed between the VSTs at not only 63Hz but at 125Hz on the fingertip, and each of the corresponding Pb-B levels. A similar tendency was also observed at the two frequencies on the toe. The measurement of VSTs was considered to be an effective screening test for sensory nerve disorders caused by lead poisoning. PMID- 7474498 TI - [The effects of long-term intake of restricted calcium, vitamin D, and vitamin E and cadmium-added diets on the bone mass of mouse femoral bone: a microdensitometrical study]. AB - Effects of undernutrition on development of cadmium (Cd) poisoning were investigated in mice over a long experimental period ranging from 29 days to 24 months. The study was designed to examine low in calcium, low in vitamin D and low in vitamin E diets, which were considered neither to induce lesions of Ca, vitamin D and vitamin E deficiency nor in nutrition. Twenty-nine-day-old female ICR strain mice were separated into 7 groups and different diets were given to each group. Group I a commercial diet (Intact), Group II a low in Ca and low in D diet, Group III a low in Ca, low in D and low in E diet, Group IV a low in Ca, low in D and 20ppm Cd diet, Group V a low in Ca, low in D and 40ppm Cd diet, Group VI a low in Ca, low in D, low in E and 20ppm Cd diet and Group VII a low in Ca, low in D, low in E and 40ppm Cd diet. Using X-rays of the hind legs, the bone density was measured at the middle of the femur employing a densitometer; effects of the undernutrition and the cadmium-added diet on bone density were investigated referring to five indices: 1) peaks of the density of the middle of the bone cortex (h1, h2) (the maximum density of the bone cortex (h1 + h2)/2 [hmax]), 2) the lowest density of the middle of the bone marrow (h3) (the minimum density of the bone marrow [hmin]), 3) the bone cortical width (d1 + d2), 4) the bone marrow width (d) and 5) the cortical bone index (the ratio of cortical bone width to bone width. [CBI]). The long-term effects of cadmium. a) Comparisons among groups II, IV and V.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7474499 TI - [The effects of long-term intake of restricted calcium, vitamin D, vitamin E and cadmium-added diets on the skeletal muscles of mice: an enzyme histochemical study]. AB - To clarify the effect of chronic cadmium exposure on muscles, 29-day-old female ICR strain mice were separated into 7 groups and various diets were given to each group. Group I a commercial diet (Intact), Group II a diet low in calcium and low in vitamin D, Group III a diet low in Ca, low in D and low in vitamin E, Group IV a low in Ca, low in D and 20ppm cadmium-added diet, Group V a low in Ca, low in D and 40ppm Cd diet, Group VI a low in Ca, low in D, low in E and 20ppm Cd diet and Group VII a low in Ca, low in D, low in E and 40ppm Cd diet. The levels of vitamin D and vitamin E were designed to be low in each diet but their amounts fulfilled the minimum nutritional requirements. The experimental period was long, ranging from 29 days to 24 months. Using a microscope, two skeletal muscles, the soleus (red muscle) and the gastrocnemius (white muscle) of mice with chronic Cd effects were observed after staining with H. E. and analyzed after enzyme histochemical reaction to ATPase. Using ATPase stain, type I muscle fibers were distinguished from type II muscle fibers. The long-term effects of cadmium. 1) Comparisons among groups II, IV and V. After 18 months it was found in group IV that myopathic muscle damage (increased random variation of fiber size [Size] and widening of interstitial tissue [Widening]) were more prominent that in group II. In group V myopathic muscle damage (Size and Widening) and type II atrophy were more prominent that in group IV. At 24 months it was found in group IV that myopathic muscle damage (Size, internal nuclei [IN] and Widening) and type I and type II atrophy were more prominent than in group II. In group V myopathic muscle damage (Size, IN and Widening) and type I and type II atrophy were more prominent than in group IV. 2) Comparisons among group III, VI and VII.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 7474500 TI - Seasonal fluctuations of Dermatophagoides mite population in house dust. AB - To evaluate seasonal fluctuations of Dermatophagoides species (sp.) in residential houses in Mie Prefecture, Japan, we employed the ELISA inhibition method with rabbit polyclonal antibody to eggs and adult mites. House dust accumulated for seven days in vacuum cleaners was collected from 14 houses of atopic patients every month for one year. The one-week-dust samples weighed from 2.5 to 117 g (mean 35.9 g). The density of Dermatophagoides spp. in the dust samples were 5 to 755 mites per gram (mean 109). During one week, 100 to 20,400 (mean 4,200) mites of Dermatophagoides sp. were collected. Among 142 samples in one year, 50 (35.2%) contained more than 100 mites per gram, which is a critical number for atopic symptoms. From December to February, 12 (34.5%) out of 35 samples contained more than 100 mites per gram. These results suggest that Dermatophagoides sp. have become a year-round allergen in Japan. PMID- 7474501 TI - Induction of IgE antibody production in mice with different DPT-vaccine preparations. AB - Antibody production in mice after immunization with diphtheria-pertussis-tetanus (DPT) vaccine was investigated. Six lots of the vaccine produced in the same year by six manufacturers in Japan were chosen. Production of IgE antibody specific to either diphtheria or tetanus toxoid varied among vaccine preparations used, although there was no apparent difference in IgG antibody production after immunization. In addition, the level of IgE antibody specific to diphtheria toxoid was correlated with that of IgG1 antibody and inversely with that of IgG2 antibody. These results suggest that each vaccine preparation may induce a distinct pattern of antibody production and, therefore, the type of immune response induced by vaccination may vary among vaccine preparations. PMID- 7474502 TI - Apoptosis of HIV-infected cells following treatment with Sho-Saiko-to and its components. AB - Baicalein and baicalin are components of Sho-saiko-to (SST), a Chinese medical drug which is claimed to be therapeutically effective in treating HIV-infected patients. Although 20 micrograms/ml of baicalin was not cytotoxic to CEM cells, a cultured T cell line, it proved to be cytotoxic to HIV-infected CEM cells (CEM HIV) with a higher HIV-releasing capacity and DNA fragmentation was detected within 24 hr of incubation. However, after incubation of CEM-HIV with a lower dose of baicalin (0.1, 0.3 and 2 micrograms/ml) for 24 and 48 hr, the viable cell number increased by about 25% and the p24 release into the medium was 25% lower than that of the control. After further incubation in the presence of the agent for 6 and 9 days, only cells with a lower HIV-releasing capacity survived. Baicalin might selectively induce apoptosis of CEM-HIV cells which have a high virus-releasing capacity, and stimulate proliferation of CEM-HIV which have a relatively lower capacity of HIV-production. PMID- 7474503 TI - Mechanism of chloramphenicol-induced modulation of mouse ileal motility. AB - The effect of chloramphenicol (CAP) on the intestinal motility of mice was studied. Acute and chronic CAP treatment significantly increased the food transit time. CAP produced concentration-dependent inhibition of motility of the isolated ileum of mice. Prazosin, propranolol, atropine, ouabain and chlorpromazine all failed to modulate or counteract the CAP-induced inhibition of ileal motility. However, naloxone and hexamethonium slightly modified the inhibitory response of CAP. The inhibitory response of CAP was markedly counteracted by cystine, a guanylate cyclase inhibitor. CAP increased the activity of Ca(++)-ATPase in the ileum in all experiments. Our results suggest that the CAP-induced inhibition of the intestinal motility is not mediated through adrenergic, cholinergic and cAMP or through inhibition of the electrogenic pump. Compared to thiamphenicol (TAP), CAP, with a p-NO2 group in its structure, exhibited more pronounced alteration of both intestinal motility and Ca(++)-ATPase activity. We, therefore, suggest that greater inhibition of ileal motility induced by CAP is possibly a p-NO2-cGMP Ca(++)-ATPase-mediated mechanism. PMID- 7474504 TI - [Nephritogenic immune response]. PMID- 7474505 TI - [Exercise intensity and renal hemodynamics]. AB - This study investigated the effects of exercise intensity on renal hemodynamics. Three healthy male subjects underwent exercise tests on a bicycle ergometer at 7 different work loads for 15 min. The indicators of exercise intensity employed in this study were the percentage of maximal oxygen uptake (%VO2max) and percentage of ventilatory threshold (%VT). As renal clearance parameters, para aminohippurate clearance (CPAH) and inulin clearance (CIN) were measured by the continuous infusion technique. Indicators of renal hemodynamics during exercise were the percentage of CPAH, CIN and filtration fraction (FF) at rest. Plasma norepinephrine (NE), plasma epinephrine (E) and plasma renin activity (PRA) were measured. The best regression models between renal clearance parameters and exercise intensity were selected using Akaike's information criterion (AIC). 1) The renal clearance parameters used during exercise were %CPAH, which determined the cubic regression model and %CIN, which determined the quadratic regression model as the exercise intensity increased using AIC. 2) The percentage of maximal oxygen uptake at the onset of decrease in %CPAH and %CIN were 35 and 49%VO2max, respectively, while %CPAH and %CIN began to decrease at 75 and 105%VT, respectively. Intensity of exercise at the onset of increase in %FF was 49%VO2max or 106%VT. 3) The renal clearance parameters during exercise decreased linearly as NE, E and PRA increased. The increase in NE contributed mainly to a change in renal clearances shown by multiple regression analysis. The above results suggest that the relationship among renal plasma flow (RPF), glomerular filtration rate (GFR) and exercise intensity were demonstrated by the cubic regression model and quadratic regression model using AIC, respectively.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7474506 TI - [Treatment of secondary hyperparathyroidisms of predialysis chronic renal failure with low doses of 1.25(OH)2D3]. AB - Although disorders of renal calcitriol synthesis play an important role in the pathogenesis of secondary hyperparathyroidism in the early and moderate phase of chronic renal failure, the treatment of secondary hyperparathyroidism with vitamin D metabolite has not attained consensus from the view point that is accelerates the progression of renal disease. The aim of this study was to evaluate the efficacy and adverse effect of low-dose daily oral treatment of 1.25 vitamin D for patients with mild to moderate renal failure. Fifteen chronic renal failure patients with serum creatinine ranging from 2.5 to 6.1 mg/dl and serum intact parathyroid hormone ranging from 100 to 450 pg/ml, were treated with oral 0.25-0.5 micrograms of 1.25 vitamin D for six months, after a six month control periods. In the six months control periods, serum intact parathyroid hormone, alkaline phosphatase activity, and bone gla protein increased significantly, however after the treatment of 1.25 vitamin D, serum intact parathyroid hormone, alkaline phosphatase activity, and bone gla protein decreased significantly. Serum calcium concentration increased significantly after the initiation of 1.25 vitamin D treatment, so it could not be ascertained whether or not 1.25 vitamin D directly suppressed parathyroid hormone synthesis. Bone mineral densities did not change within one year. Renal function was evaluated from the slopes of the reciprocal serum creatinine concentration versus time. The slopes did not change after the administration of 1.25 vitamin D. In conclusion, 1.25 vitamin D treatment of secondary hyperparathyroidism in patients with mild to moderate renal failure had beneficial therapeutic effect on humoral bone parameters, and did not show any adverse effect on renal function. PMID- 7474507 TI - [Relationship between vasoactive substances and changes in blood pressure during hemodialysis]. AB - In order to clarify the relationship between vasoactive substances and changes in blood pressure (BP) during hemodialysis (HD) in patients with end-stage renal disease, we measured plasma renin activity (PRA), plasma concentration of aldosterone (PAC), and the plasma concentrations of epinephrine (E), norepinephrine (NE), dopamine (DA) and atrial natriuretic peptide (ANP) in patients on HD. Fourty-eight patients, consisting of 24 males and 24 females, were included in this study. Their mean age was 63.8 years, and their mean HD duration was 55.9 months. In 9 patients without diabetes mellitus (DM) and whose BP was stable during HD, PRA and plasma concentrations of E and NE increased significantly during HD, and that of DA decreased during HD. In 9 patients without DM and whose BP fell during HD, PRA and plasma concentrations of E and NE showed no significant response to the decrease in body weight during HD. In spite of the increase in NE concentration during HD in 8 patients without DM and whose BP was usually hypotensive, BP remained low. This might be due to the decrease in sensitivity of their peripheral autonomic receptors to NE. In the 9 patients with DM whose BP was stable and 13 patients with DM whose BP fell during HD, vasoactive substances made almost no effective response to the decrease in body weight during HD. In conclusion, we must take into consideration the fact that both hypotensive patients during HD and diabetic patients on HD might exhibit an abnormal response of their vasoactive substances. PMID- 7474508 TI - [Binding of IgA1 to monocyte/macrophage cell lines (THP-1, U937) in IgA nephropathy--a possible role of O-glycan in the IgA1 molecule]. AB - IgA1 is an exceptional serum glycoprotein because it has O-glycans in the hinge region. It has been observed that the monocyte/macrophages infiltrate within the glomeruli in IgA-N. On the other hand, it is well established that the carbohydrate side chains (N-glycans) of the IgG molecule play a role in the binding of IgG to Fc receptors. Therefore, the binding of IgA1 to monocyte/macrophage cell lines was observed with the aim of clarifying the role of the O-glycan side chains in IgA-N in the initial mechanism of glomerular damage due to the interaction between the O-glycan chains on the IgA1 molecule and infiltrating monocyte/macrophages. Two human myelomonocytic cell lines, THP-1 and U-937, were activated and incubated with separated IgA1 (IgA-N, n = 16; other glomerulonephritides (other GN), n = 15; healthy controls, n = 9). The binding attitude of IgA1 to the cell lines was observed by flow immunofluorometry using a FACScan. FACScan showed that the binding of IgA1 to both of the stimulated monocyte/macrophage cell lines was increased in IgA-N compared to the normal controls and other GN. The binding of IgA1 to THP-1 was partially, but definitely inhibited by adding 100 mM melibiose (19.6 +/- 7.7%) and galactose (13.1 +/- 2.9%), but not glucose (2.9 +/- 2.2%), lactose (4.7 +/- 4.7%) and mannose (3.3 +/ 3.3%). These results suggested that THP-1 had a receptor that recognized the O glycan in the IgA1 hinge region.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7474509 TI - [Long-term hemodialysis treatment using femoral vein puncture method (FV-method) as blood access in 12 patients]. AB - It is well known that blood access is essential for long-term hemodialysis treatment. Arteriovenouos fistula (AVF) is the most widely used method. However, this method of access frequently fails (access failure) as a result of stenosis. We attempt simple femoral vein puncture (FV-method) instead of AVF in such patients and have experienced 12 patients who were undergoing hemodialysis treatment using the FV-method, three times a week for more than one year. We devised special needles (18- and 19-gauge) for the FV-method. Generally, we use a 19-gauge needle with 4 side holes. We discuss here the results of 12 patients consisting of 4 males and 8 females with a mean age of 57.9 years, a mean duration of dialysis of 10.0 years, and a mean duration of FV-method of 3.5 years. Their underlying diseases were chronic glomerulonephritis (9 patients), diabetic nephropathy (2 patients) and nephrosclerosis (1 patient). Before the use of the FV-method, AVFs were attempted a man of 3.8 times and an artificial graft, 4 times in 3 patients. Ten patients were outpatients and 2 were inpatients. As for the indications of the FV-method, 11 patients had access failure and another had suffered from heart failure resulting from an over flow of blood through AVF. KT/V, PCR and TACBUN were measured monthly and were within the normal range in almost all of the patients. Concerning complications of the FV-method, hematoma formation after detachment of the needle at the end of dialysis and pain at needle puncture were sometimes noted.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7474511 TI - [Endocrinological analysis before and after living-related renal transplantation in a patient of Bartter's syndrome]. AB - In order to clarify endocrinological changes before and after living-related renal transplantation in a patient of Bartter's syndrome involving chronic renal failure, serial quantitative determinations of the renin-angiotensin-aldosterone system and 24-hour urinary excretion of 6-keto-prostaglandin F1 alpha and kallikrein were performed. A male patient was admitted to hospital because of a pale face and short stature at the age of 13 years. He was 126 cm in height (M 3.8SD). Blood pressure was 110/60 mmHg and edema was not observed. Laboratory findings showed that his hematocrit was 22.1%, serum potassium 2.9 mEq/1, creatinine clearance was 30.7 ml/min/1.73m2 and beta 2-microgobulin was elevated to 39.9 mg/1 in urinalysis. Plasma renin activity and aldosterone were remarkably elevated to 24.23 ng/ml/hr and 738 pg/ml, respectively. The kidney biopsy specimen showed diffuse glomerulosclerosis and hypertrophic change of the juxtaglomerular apparatus was also demonstrated. He was diagnosed as Bartter's syndrome with short stature and chronic renal failure. At the age of 18, he was introduced on hemodialysis and the living-related renal transplantation was performed the next year. Two weeks after the transplantation, plasma renin activity, angiotensin, I,II and aldosterone were markedly changed from 37.8 to 2.3 ng/ml/hr, 2400 to 220 pg/ml, 256 to 17 pg/ml and 3700 to 110 pg/ml, respectively. Urine prostaglandin F 1 alpha was improved from 860 to 321 ng/day and kallikrein was also changed from 400 to 25.2 micrograms/day. These results indicated that abnormalities of several hormones in Bartter's syndrome could be normalized by living-related renal transplantation.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7474510 TI - [Clinical analysis of 14 patients with anti-myeloperoxidase antibody positive rapid progressive glomerulonephritic syndrome]. AB - We investigated the clinical features and outcome of 14 patients with anti myeloperoxidase antibody (MPO-ANCA) positive rapidly progressive glomerulonephritic syndrome. Underlying diseases included microscopic polyarteritis in 6 patients, idiopathic crescentic glomerulonephritis with lung hemorrhage in 2 patients, idiopathic glomerulonephritis in 3 patients, rapidly progressive glomerulonephritic syndrome without renal biopsy in 1 patient, crescentic glomerulonephritis associated with Sjogren syndrome and progressive systemic sclerosis in 1 patient and crescentic glomerulonephritis associated with sarcoidosis in 1 patient. Five patients were male (mean age, 59.2 years) and 9 were female (mean age, 54.0 years). On admission, most patients had anemia, leukocytophilia, and marked elevation of C-reactive protein (CRP). Average hemoglobin, white blood cell count and CRP levels on admission were 8.1 mg/dl, 11,500/mm3 and 14.7 mg/dl, respectively. Average serum creatinine was 4.0 mg/dl. All patients were treated with steroids either with or without cyclophosphamide. As the patients recovered clinically, the MPO-ANCA titers declined. Although most patients responded well to immunosuppressive therapy, some died of serious complications (such as acute respiratory distress syndrome, fungal infection, and pneumocystis pneumonia). The prognosis of patients with severe renal failure was especially poor. We conclude that early diagnosis, treatment and intensive care during immunosuppressive therapy are very important in the management of MPO-ANCA positive rapidly progressive glomerulonephritis. PMID- 7474512 TI - [Glomerular lesion in patients with type II mixed cryoglobulinemia having antibodies to hepatitis C virus]. AB - We report here two cases of mixed cryoglobulinemia showing renal involvement associated with hepatitis C virus (HCV) infection. The subjects were 62- and 63 year-old males. Both patients presented with purpura on the legs, which was diagnosed as allergic vasculitis by skin biopsy. Case 1 followed a clinical course of progressive nephrotic syndrome with mild hematuria. He also had diabetes mellitus and hypertension. In contrast, case 2 showed only mild hematuria without proteinuria at the time of the renal biopsy. Both cases had immunological disarrangements, such as severe hypocomplimentemia and seropositive rheumatic factor. Recently, it was reported that patients with type II mixed cryoglobulinemia had HCV seropositivity, and revealed membranoproliferative glomerulonephritis. These facts strongly suggested that renal lesions are the result of direct damage mediated by cryoglobulinemia and an activated complement pathway through an immune complex mechanism related to HCV. PMID- 7474513 TI - [A case of renovascular hypertension with nephrotic syndrome]. AB - A 68-year-old male patient with renovascular hypertension (RVHT) and nephrotic syndrome (NS) is described. He was admitted to our hospital for detailed investigation of severe hypertension and massive proteinuria. After admission, a diagnosis of RVHT with a right non-functional kidney and NS was made. Nephrectomy and contralateral renal biopsy were performed for refractory hypertension and detailed investigation of the NS, respectively. The renal biopsy showed focal segmental glomerulosclerosis (FGS) in the left kidney, whereas the nephrectomised kidney exhibited only ischemic change. After the operation, his blood pressure became stable without anti-hypertensive agents, but proteinuria remained in the nephrotic range. Six months later, proteinuria had disappeared and his renal function was stable. These findings suggest that NS and FGS might have resulted from an activated renin-angiotensin-axis and that the prolonged NS was due to severe glomerular injury. Although there have been many reports describing the relationship between RVHT and FGS in an experimental environment, this relationship is very rare in clinical cases. Therefore we present this case to increase understanding of the cause of FGS. PMID- 7474514 TI - [Studies on functions of neutrophils in MPO-ANCA related glomerulonephiritis and plasma effect on MPO release from neutrophils]. AB - In order to examine the role of myeloperoxidase (MPO) and neutrophil activation in the pathogenesis of MPO-ANCA related necrotizing crescentic glomerulonephritis (GN), we investigated MPO release and superoxide anion (O-2) production from the neutrophils in MPO ANCA related GN. Plasma effect on MPO release from neutrophils was also studied in MPO-ANCA related GN. Neutrophils and plasma were obtained from 10 patients with MPO-ANCA-related GN (A), 10 patients with MPO-ANCA non related GN (B) and 10 healthy controls (C). MPO release and O-2 production were assayed by using chemotactic factor, N-formyl-methionyl-leucyl-phenyalanine (FMLP) and cytochalasin B (CB). In MPO-ANCA related GN, MPO release and O-2 production were significantly higher than in the other groups. [MPO release (%) A: 43.02 +/- 18.33, B: 33.26 +/- 11.58, C: 19.73 +/- 8.38, sensitivity to FMLP (%) A: 894.56 +/- 381.96, B: 512.86 +/- 314.91, C: 301.12 +/- 139.8, O-2 production (nmoles/min/10(6) cells) A: 13.06 +/- 5.28, B: 3.57 +/- 2.02, C: 2.83 +/- 1.16] Furthermore, MPO release, sensitivity to FMLP and O-2 production were shown to increase in parallel with clinical disease activity. Plasma from MPO ANCA related GN increased sensitivity to FMLP in neutrophils obtained from healthy controls [plasma concentration 1%, A: 311.5 +/- 110.3, B: 227.2 +/- 101.4, C: 215.5 +/- 93.7], and plasma from MPO-ANCA related GN suppressed %MPO release from neutrophils with MPO-ANCA related GN [plasma concentration 0.1%, A: 11.56 +/- 6.17, C: 15.51 +/- 8.01]. It was concluded that in patients with MPO ANCA related GN, large amounts of MPO can be easily released from activated neutrophils and this might be influenced by some stimulating and suppressing factors contained in the plasma. PMID- 7474515 TI - [Agents preventing decrease in ultrafiltration in a rat model with peritoneal hyperpermeability]. AB - Ultrafiltration failure has been one of the major causes of drop out from CAPD treatment. The present study was designed to develop an ultrafiltration failure model in the rat and to assess whether various agents could prevent a decrease in ultrafiltration capacity in this rat model. Peritoneal hyperpermeability was induced by repeated intraperitoneal injection of 15 ml of 4.25% dextrose dialysate for 7 days. Some of the rats received agents, such as phosphatidylcholine, chondroitin sulfate, siliac acid, heparan sulfate, keratan sulfate, sodium sulfate, sodium phosphate, and heparin sodium, simultaneously administered with 4.25 % dextrose dialysate for the same duration. Ultrafiltration volume, D4/D0 ratio of glucose and peritoneal net fluid absorption were evaluated by 4-hour dwelling of 30 ml of 2.5% dextrose dialysate. Sodium phosphate and heparan sulfate prevented peritoneal hyperpermeability due to repeated injection of hypertonic dialysate. Sodium phosphate, sodium sulfate, heparin and heparan sulfate suppressed peritoneal net fluid absorption, resulting in an increase in ultrafiltration. These findings may be applicable to CAPD patients with ultrafiltration loss. PMID- 7474516 TI - [Role of renal thromboxane in salt-sensitive blood pressure rise in DOCA salt hypertension]. AB - Urinary excretion and release of prostaglandins (PG) from isolated glomeruli and renal papilla of deoxycorticosterone acetate (DOCA)-treated rats fed with a normal salt (0.6% NaCl) diet and a high salt (4%NaCl) diet were determined. Mean blood pressure was significantly higher in the high salt diet group than in the normal salt diet group (146.2 +/- 2.3 vs 118.6 +/- 1.9 mmHg, p < 0.01). Urinary excretion of thromboxane (Tx) B2, stable metabolite of Tx A2, and 6-keto PG-F1 alpha, a stable metabolite of prostacyclin, increased significantly in the high salt diet group compared to the values of the normal salt diet group increased significantly by 104%, 55%, and 74% compared to those of the normal salt diet group, respectively. Release of 6-keto-PG F1 alpha from renal papilla of the high salt diet group decreased significantly, but there were no intergroup differences in the release of PG E2 and Tx B2. Stepwise multiple linear regression analysis showed that the significant contributory factors underlying the mean blood pressure were urinary excretion of Na (F = 14.187, p < 0.01) and release of Tx B2 from isolated glomeruli (F = 4.135, p < 0.05). These findings suggest that the manifestation of Tx synthesis in renal glomeruli has a predominant role in the salt sensitive pressor response of DOCA salt hypertension in rats. PMID- 7474517 TI - [Role of Helicobacter pylori in gastro-duodenal mucosal lesions in patients with end-stage renal disease under dialysis treatment]. AB - It is known that Helicobacter pylori (H. pylori) plays an important role in gastritis and peptic ulcer disease in the general population. Although upper gastrointestinal mucosal lesions have been one of the most common complications in patients with chronic renal failure, quite few reports are available regarding the prevalence of H. pylori and its influence on the upper gastro-intestinal tract. The present study was conducted to examine whether H. pylori is involved in the pathogenesis of upper gastrointestinal mucosal lesions in dialysis patients. The subjects consisted of 43 dialysis patients with upper gastro intestinal tract symptoms. Thirty-four patients without any known kidney disease were used as controls. Gastric mucosa and gastric juice were obtained endoscopically. For the determination of H. pylori, culture of biopsy specimens from the gastric mucosa and histopathological examination with hematoxylin-eosin stain were used. Concentrations of serum gastrin and gastric juice ammonia were also measured. H. pylori was observed in 53.5% of the dialysis group and 64.0% of the controls. Gastro-duodenal lesions in H. pylori-positive dialysis patients included atrophic gastritis, superficial gastritis, erosive gastritis, and gastric ulcer. In the dialysis group, ammonia concentrations in the gastric juice were higher in patients with H. pylori than in those without H. pylori (489.1 +/- 35.8 mu g/ml vs 67.0 +/- 19.2 mu g/ml, p < 0.001). The former value was also higher than that seen in the H. pylori-positive controls (152.4 +/- 18.7 mu g/ml, p < 0.01). Serum levels of gastrin were significantly higher in patients with H. pylori than in patients without H. pylori (ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7474518 TI - [A new protein titrator tape for self-assessment by outpatients with proteinuria]. AB - l- have invented a new dipstick (protein titrator tape) for measuring the volume of protein excreted in the 24-hour urine. The principle of the method is based on the protein error of indicators with the modification of a conventional dipstick test. The dipstick consists of two thick filter papers, containing differently adjusted pH indicators of tetrabromphenol blue, making it possible to detect a wide range of protein concentrations in the urine using a standard color chart that includes twenty color blocks. Two hundred and ninety outpatients had their urine samples assessed with this method as well as with the pyrogallol red test as a comparative study for quantitative measurement of protein concentrations. The new-type dipstick method exhibited good correlation with the results of the pyrogallol red test, especially in the range of protein concentrations from 50 mg/dl to 400 mg/dl, showing the linear equation of "y (Pyrogallol red) = 10.5 + 0.99 x (Dipstick) (r = 0.91, P < 0.01)". Although there was good correlation with the pyrogallol red test at higher concentrations from 400 mg/dl to 1,000 mg/dl, the dipstick method tended to exhibit lower concentrations than those indicated by the counterpart method. The rate of consistency between observers was quite high. This new-type dipstick method will offer a reliable method for patients or their family to assess their protein excretion in the urine every 24 hours at home using a portable urine sampling device. PMID- 7474519 TI - [A case of nephrotic syndrome associated with severe interstitial pneumonitis due to cyclophosphamide whose life was saved by steroid pulse therapy]. AB - A 53-year-old man was admitted for treatment of nephrotic syndrome due to membranous nephropathy. Since he did not improve with prednisolone alone, cyclophosphamide was administered concomitantly. However, this induced severe interstitial pneumonitis, which was not cured by discontinuation of the drug, but disappeared completely with six episodes of methylprednisolone pulse therapy. The incidence of cyclophosphamide-induced interstitial pneumonitis is low, but the mortality is high. Since cyclophosphamide is often used for the treatment of nephrotic syndrome, attention should be focused on the incidence of interstitial pneumonitis. Discontinuation of cyclophosphamide and intensive therapy are required when it occurs. PMID- 7474521 TI - Role of hepatic receptors in controlling body fluid homeostasis. PMID- 7474520 TI - [A case of hemolytic uremic syndrome associated with circulating anticoagulant]. AB - A previously well 66-year-old woman was admitted to our hospital because of severe abdominal pain, oliguria, jaundice and hemoglobinuria. At admission, the following values were obtained: red blood cell count 261 x 10(4)/mm3, serum creatinine 2.8mg/100ml, total bilirubin 8.0mg/ml, indirect bilirubin 5.7mg/100ml, LDH 13886 WU, negative direct and indirect Coomb's test and peripheral blood smear showing schistocytosis. She was diagnosed as hemolytic uremic syndrome and was successfully treated with plasma infusion, plasma exchange and hemodialysis. The results of the tests for verotoxin titer of E. coli O157: H7 and circulating anticoagulant were positive. The test result for anticoagulant eventually became negative during her clinical course. In this case, circulating anticoagulant might have contributed to the pathogenesis of hemolytic uremic syndrome. PMID- 7474522 TI - Effects of Ca2+ and Mg2+ on dynamics of the polar head group of phosphatidylserine bilayers. AB - The effects of Ca2+ and Mg2+ on the molecular motion of the polar head group in phosphatidylserine (PS) bilayers were measured by the time-resolved fluorescence depolarization method probed by 1,2-dihexadecanoyl-sn-glycero-3-phospho-[N-(4 nitrobenzo- 2-oxa-1,3-diazole)]ethanolamine [formula: see text] (NBD-PE). By this method, the rate and width of the molecular motion at the fluorescent moieties in the probe molecules could be evaluated as the wobbling diffusion rate (Dw, s-1) and the half cone angle of the wobbling cone (theta c, degree). The values of Dw and theta c measured for NBD-PE embedded in bovine brain phosphatidylserine bilayers were 3.7 x 10(7) s-1 and 46 degrees in the absence of divalent cations at 25 degrees C. When 3 mM of Ca2+ was added, both Dw and theta c distinctly dropped to 1.7 x 10(7) s-1 and 38 degrees, respectively. By the addition of 3 mM of Mg2+, however, only Dw decreased to 2.7 x 10(7) s-1 and theta c remained unchanged. These results show that both Ca2+ and Mg2+ decrease the rate of motion at the head part in PS molecules, but only Ca2+ narrows the distance between the neighboring head groups. Since Mg2+ does not promote vesicle fusion, it appears that the deformation at the head group region in the bilayer structure induced by Ca2+ is an important step in the membrane fusion process. PMID- 7474523 TI - Effects of alpha-adrenergic activation on the shift of electrolytes and fluid after hemorrhage in rats. AB - To assess the effects of alpha-adrenergic activation on hemorrhage-induced shifts of K+, Na+ and fluid between the intravascular and extravascular spaces, we continuously measured changes in plasma concentrations of K+ and Na+ (delta[K+]p, delta[Na+]p) and blood volume (delta BV) over 60 min after hemorrhage in nephrectomized rats. Hemorrhage was conducted over 5 min at the level of 0.5, 1.0 and 1.5% of body weight (H0.5, H1.0, H1.5), and the result was compared with hemorrhage of 1.0% body weight after administration of alpha 1-adrenoreceptor antagonist prazosin (0.2 mg/kg, H1.0P). delta[K+]p increased significantly (p < 0.05) after hemorrhage, and the peak value was proportional to the level of hemorrhage (0.18 +/- 0.08, 0.62 +/- 0.22 and 1.64 +/- 0.19 meq/l in H0.5, H1.0 and H1.5, respectively). delta[Na+]p decreased significantly (p < 0.05) after hemorrhage, and the decrease was sustained until the end of the experiment (-0.8 +/- 0.6, -1.0 +/- 0.5 and -2.2 +/- 0.5 meq/l in H0.5, H1.0 and H1.5, respectively). In H1.0P, the increase in delta[K+]p (0.25 +/- 0.09 meq/l at the peak) and the decrease in delta[Na+]p (-0.2 +/- 0.1 meq/l at the bottom) were suppressed significantly (p < 0.05) compared to H1.0. Although delta BV was greater in H1.0P than H1.0, plasma K+ content was not different between the groups. In H1.0, the calculated concentrations of K+ and Na+ in the fluid which shifted into the intravascular space ([K+]sf, [Na+]sf) in the first 30 min after hemorrhage were higher in [K+]sf (6.25 +/- 0.70 meq/l) and lower in [Na+]sf (128.0 +/- 3.2 meq/l) than the pre-hemorrhage plasma level. With regard to H1.0P, [K+]sf and [Na+]sf were not different from the pre-hemorrhage level of plasma. These results suggest that alpha-adrenergic activation after hemorrhage induces K+ movement into plasma to increase [K+]p, which might be related to the fluid shift from the intracellular to the extracellular space. PMID- 7474524 TI - Effects of inhibitors of ouabain-sensitive Na+, K(+)-ATPase and Li+ ions on the neuromuscular transmission of the frog. AB - The effect of blockade of ouabain-sensitive alpha 2 and alpha 3 (neural type) isozymes of Na+, K(+)-ATPase was investigated on frog neuromuscular preparations by recording the frequency augmentation-potentiation (FAP) of the endplate potential, an electrophysiological and neuropharmacological technique to analyze the drug actions on the release process of the readily releasable transmitter quanta. Erythrosin B, which was thought to selectively inhibit the neural type Na+, K(+)-ATPase, pivoted the log-linear FAP relation counterclockwise without altering the intercept on the ordinate. Chlormadinone had a similar action. An increase in the concentration of extracellular K+ ions pivoted the FAP relation clockwise with a concomitant upward shift of the intercept on the ordinate, and low K+ Ringer's solution produced an inverse effect. In contrast, Li+ ions shifted the FAP relation upwards dose-dependently leaving its slope unchanged. Cinnarizine, a blocker for inositol-1,4,5-trisphosphate-induced Ca2+ release, and 5,5'-dimethyl-1,2-bis(2-amino-phenoxy)ethane-N,N,N',N'-tetraacetic acid, a specific intracellular Ca2+ chelator, significantly antagonized the potentiating action of Li+. The ouabain-sensitive neural type Na+, K(+)-ATPase isozyme, which is abundant in neural tissues, seems to play an important role in stimulation frequency-dependent modulation of the quantal transmitter release such as FAP. PMID- 7474525 TI - Phosphomonoesters in the guinea-pig taenia caeci: pH-dependency of the phosphomonoester peaks in 31P-NMR. AB - In the smooth muscle of guinea-pig taenia caeci, unlike cardiac and skeletal muscles, two prominent peaks are constantly observed in the phosphomonoester (PME) region (at 6.8 and 6.3 ppm) of 31P-NMR spectra. Tissue extracts, whose pH were adjusted to the intracellular pH of 7.1, also showed such peaks. According to the pH-dependency of the chemical shifts of known PMEs, these peaks were identified as phosphorylethanolamine (PEt) and phosphorylcholine (PCh), the intermediary metabolites of membrane phospholipid turnover. In normal solution, the intracellular pH (pH(i)) which was estimated from the chemical shifts of PMEs (PEt 7.07; PCh 7.17) agreed with that from P(i) (7.10). When preparations were exposed to hypoxia and high-Mg2+ conditions, a reasonable correlation was found between estimations of pH(i) from the PME and P(i) peaks. The chemical shifts of these PME peaks may be used for estimation of pH(i), at least, when there is a need to know relative changes of pH(i), as a complementary purpose, and also whenever the P(i) peak is not clearly observed. PMID- 7474526 TI - Hemodilution-induced inhibition of cardiovascular responses to some vasoactive agents in anesthetized cats. AB - Cardiovascular responses to adrenaline and acetylcholine (ACh) were investigated in anesthetized, artificially ventilated cats in control and after induction of acute normovolemic hemodilution. Progressive replacement of blood by high molecular weight dextran was performed in three steps of 20% each of the total estimated blood volume. Hemodynamic responses were recorded at four stages: the control stage and after the 1st, 2nd, and 3rd exchanges of blood for dextran. With the fall in hematocrit (Ht) there was a corresponding significant (p < 0.05) increase in heart rate (HR), cardiac output (CO), and stroke volume (SV), and a decrease in systemic vascular resistance (TPR). However, left ventricular systolic pressure (LVSP), left ventricular contractility (LV dP/dtmax), mean arterial pressure (MAP), and right atrial pressure (RAP) did not show any significant (p > 0.05) change due to hemodilution. The cardiovascular responses of intravenously administered adrenaline and ACh were significantly (p < 0.05) attenuated. Responses to sodium nitroprusside (SNP), a potent vasodilator and an exogenous source of nitric oxide, were also attenuated after hemodilution. The increase in SV and HR seem to be the contributing factors to the CO response. Our results indicate that the cardiovascular responsiveness to adrenaline, ACh and SNP is reduced during acute hemodilution which could be due to inadequate myocardial and vascular O2 supply. The possibility of a modulatory role of an endothelium-dependent mechanism and reflex regulatory responses by arterial baroreceptors during hemodilution also exists. PMID- 7474527 TI - GABA-induced chloride current in catfish horizontal cells mediated by non-GABAA receptor channels. AB - GABA-induced currents were recorded in cone-driven horizontal cells dissociated from the catfish, using the patch clamp technique in the whole-cell configuration. GABA-induced current consisted of two components; approximately 80% of the current was blocked by 100 microM picrotoxin (PTX), and the remaining 20% of the current was blocked when extracellular Na+ was replaced with Li+ or choline. The PTX-sensitive current was carried by Cl-. When methanesulfonate was substituted for the intrapipette Cl-, the reversal potentials of the PTX sensitive current shifted to a more negative potential close to the Cl- equilibrium potential. The PTX-sensitive current was unaffected either by bicuculline (up to 500 microM), pentobarbital (100 microM), or diazepam (100 microM). These observations suggest that the PTX-sensitive current is not mediated via GABAA receptors. Baclofen, a GABAB receptor agonist, had no effect, suggesting the absence of GABAB receptors. Cis- or trans-4-aminocrotonic acid (CACA and TACA), GABAC receptor agonists, evoked currents in a dose-dependent manner. The potency sequence was TACA > GABA > muscimol > CACA. These observations suggest that the PTX-sensitive current of cone-driven horizontal cells is carried via the GABAC receptor channel. The Cl- equilibrium potential in intact cells was estimated to be approximately -30 mV by recording the GABA induced voltage response of dissociated cells with a conventional intracellular microelectrode. The intracellular Cl- concentration seemed to be approximately 40 mM. From these results, it is suggested that GABAC receptor channels shape the kinetics of light-induced responses of horizontal cells, and mediate a chemical coupling between neighboring horizontal cells. PMID- 7474528 TI - Actions of nipecotic acid and SKF89976A on GABA transporter in cone-driven horizontal cells dissociated from the catfish retina. AB - Whole-cell voltage-clamp recordings were made from dissociated horizontal cells of the catfish retina. In the presence of picrotoxin (PTX, 100 microM), GABA evoked a sustained inward current at negative holding potentials. Dose-response measurements were well fitted by a logistic curve with a Hill coefficient of 1.11 and EC50 of 9.76 microM. When external Na+ was replaced with Li+, this GABA induced current was eliminated. The substitution of methanesulfonate for Cl- also suppressed the current. This current was blocked by either nipecotic acid or SKF89976A. However, the mechanisms by which these drugs suppress the GABA-induced current differ. Intracellularly applied SKF89976A blocked the GABA-induced current, while nipecotic acid intracellularly had no effect. beta-Alanine at concentrations greater than 1 mM exerted a slight inhibitory effect. Extracellularly applied SKF89976A produced no current by itself but suppressed GABA-induced currents. Dose-response curves showed that SKF89976A has an IC50 of 0.93 microM and a Hill coefficient of 2.68. Nipecotic acid evoked a current response, like GABA. A Hill coefficient was 1.64 and an EC50 was 7.69 microM. This nipecotic acid-induced current was blocked by substituting Li+ for Na+ or by the addition of SKF89976A. This result is consistent with other studies indicating that nipecotic acid is transported in place of GABA. Extracellular Na+ was required for the prolonged suppression by extracellularly applied SKF89976A, while the extracellular Cl- depletion has no influence on the suppression. The pharmacological profile of this GABA transporter fits the neuronal rather than the glial type of cloned transporters. PMID- 7474529 TI - Parasympathetic reflex salivary secretion in the cat parotid gland. AB - The aim of the present study was to compare and to characterize the secretory and vasodilator effects in the parotid gland of sympathectomized cats elicited reflexly by electrical stimulation of the central cut ends of the chorda tympani (or glossopharyngeal) nerves (physiologically conveying gustatory stimuli), vagus nerve (visceral stimuli), and the lingual nerve after cutting the chorda tympani nerve (somatic stimuli). The threshold stimulus-intensity needed to elicit the salivary secretion and a blood flow increase was markedly lower for stimulation of the chorda tympani nerve and glossopharyngeal nerve (2 V for each nerve) than for the lingual nerve, inferior alveolar nerve, and vagus nerve (10 V for each nerve). The salivary and vasodilator responses to stimulation of each afferent nerve were all abolished by prior treatment with hexamethonium at a dose of 5 mg/kg, i.v. Prior treatment with scopolamine completely abolished the salivary secretion elicited by lingual nerve and glossopharyngeal nerve stimulation, but had less effect on the vasodilator response in the parotid gland. These results indicate that the secretory and vasodilator responses evoked by afferent traffic in the above nerves are mediated via an activation of the autonomic nervous system, particularly by a parasympathetic mechanism. They suggest that there is a marked difference in the ability of different sensory modalities to induce reflex salivary secretion and vasodilatation, there being low threshold nerve fibers in the chorda tympani and glossopharyngeal nerves and high threshold nerve fibers in the lingual nerve, vagus nerve, and inferior alveolar nerve. PMID- 7474530 TI - Neuronal nicotinic acetylcholine receptors of ganglion cells in the cat retina. AB - Ganglion cells, retrogradely labeled with DiI, were dissociated from the cat retina. Under the whole-cell clamp mode using a patch pipette, ACh induced a transient current (IACh) in all cells tested. The soma diameter of ACh-responsive cells distributed from 11 to 23 microns, indicating that the preparation included Y, X, and W cells. IACh was inward rectifying. The reversal potential of IACh was close to the zero-current potential predicted from the intra- and extracellular monovalent cation concentrations by the Goldman-Hodgkin-Katz equation. The concentration-response curve gave a Hill coefficient of 0.89 and an EC50 of 3.2 x 10(-5) M. Nicotine (10(-4) M) induced a transient current, while muscarine (10( 4) M) was ineffective. The nicotinic antagonist, hexamethonium, reduced the amplitude of IACh in a concentration-dependent manner with an IC50 of 1.3 x 10( 6) M. The muscarinic antagonist, atropine, showed a concentration-dependent reduction in the amplitude of IACh with an IC50 of 4.4 x 10(-6) M. The amplitude of IACh was dependent on the extracellular Ca2+ concentration ([Ca2+]O); small at low [Ca2+]O and large at high [CA2+]O. The time course of desensitization was also dependent on [Ca2+]O; fast at low [Ca2+]O and slow at high [Ca2+]O. The replacement of extracellular Ca2+ to Sr2+ increased the amplitude of IACh, but replacement to other divalent cations reduced it. The results indicate the presence of functional neuronal nicotinic ACh receptors in Y, X, and W cells. PMID- 7474531 TI - Augmentation and suppression of the excitatory and inhibitory neuromuscular transmission induced by the ionophores, X-537A and A23187, in the red muscle of carp, Cyprinus carpio. AB - The effects of the ionophores, X-537A and A23187, on excitatory junction potentials (ejp), inhibitory junction potentials (ijp), diphasic junction potentials (diphasic jp) composed of ejp and ijp, and miniature excitatory junction potentials (mejp) were examined in the red muscles of carp, Cyprinus carpio. When 25 microM X-537A and 5 microM A23187 were applied, the amplitude of ejp and ijp increased transiently, then decreased gradually, and finally disappeared. The duration of ejp and ijp was little affected by the ionophores. The ionophores induced a transient increase in the frequency of mejp, which then decreased gradually after the maximum increase was attained, and finally fell below the control level. The ionophores had little effect on the resting membrane potential, membrane resistance of the muscle fiber, and amplitude of the compound action potential recorded from nerve bundles innervating this muscle. The ionophores caused ACh-evoked potentials to decrease gradually and finally disappear. These results suggest that the increase in the amplitude of ejp and ijp and in the frequency of mejp might be mainly due to the augmentation of ACh release from nerve terminals caused by an elevation of the intracellular Ca2+ concentration. On the other hand, the decrease and the abolition in the amplitude of ejp and ijp and in the frequency of mejp might be mainly due to the desensitization of the postsynaptic membrane. PMID- 7474532 TI - Physical and motor fitness in twins. AB - Thirty pairs of monozygotic (MZ) and 20 pairs of dizygotic (DZ) twins, of both sexes, were studied to estimate the role of genetic and environmental factors determining fitness measurements comprising vital capacity (VC), vertical jump, flexibility, agility, and PFI score. Intra-pair variances for VC, vertical jump, and heart rate were significantly smaller (p < 0.05-0.01) in MZ than in DZ twins, where values for flexibility, agility, and PFI were insignificant. Intra-pair correlations for VC, vertical jump, and heart rate were higher (p < 0.01) in MZ than in DZ twins where the opposite was found in the case of flexibility and agility. The correlation value for PFI was higher in both MZ and DZ twins. Following adjustment for physical characteristics including age, weight, and height, none of the results of all the residual fitness measurements were similar to the unadjusted values. Both unadjusted and residual variables with smaller intra-pair variances showed higher heritability estimates. These results suggest that VC, vertical jump, and heart rate are influenced more by genetic factors than environmental factors where the inverse is true of flexibility, agility, and PFI. Thus, the genetic and environmental influence of all the fitness measurements, except VC, more or less depend upon physical characteristics in this study. PMID- 7474533 TI - Logistic characterization of left ventricular isovolumic pressure-time curve. AB - Although some investigators have attempted to express the left ventricular pressure-time curve by mathematical functions such as exponential and sinusoidal functions, none of them reasonably fits the left ventricular pressure-time curve. In the present study, we hypothesized that a ventricular isovolumic pressure-time curve could be expressed as the difference between two S-shaped curves for pressure rising and falling, and proposed a new "hybrid logistic" function to express the left ventricular isovolumic pressure-time curve. We investigated how well this hybrid logistic function fits left ventricular isovolumic pressure curves experimentally observed under physiological preload and contractility in the excised cross-circulated left ventricles of 10 dogs. The new function precisely fitted the isovolumic pressure curves regardless of preload and contractility with correlation coefficients above 0.9996, much better than the previously proposed functions. The observed values characterizing the magnitude and time course of the isovolumic pressure curve such as peak +/- dP/dt also closely correlated with the corresponding theoretical values calculated by the present best-fit function. We conclude that our new hybrid logistic function reasonably characterizes the canine left ventricular isovolumic pressure-time curve within physiological ranges of preload and contractility. The present results indicate that this hybrid logistic function is useful to evaluate left ventricular contraction and relaxation comprehensively. PMID- 7474534 TI - Effect of exercise on endotoxin shock with special reference to changes in concentration of cytokines. AB - The effects of exercise on susceptibility to endotoxin shock and the serum levels of interleukin-1 (IL-1) and interleukin-6 (IL-6) were studied in mice primed with Propionibacterium acnes (P. acnes, 1.0 mg). Endotoxin shock was induced by injection of lipopolysaccharide (LPS, 0.2 mg) 7d after the priming injection of P. acnes. The exercise-loaded mice performed voluntary exercise for 120 min on a running-wheel and LPS was injected into the tail vein immediately after the end of exercise. Two of the nine animals in the exercised group survived, whereas all the mice in the control group died. The mean survival time after LPS injection in the exercised group (10.75 +/- 4.51 h) was significantly longer than that in the control group (3.3 +/- 1.34h). The serum concentrations of IL-1 (2,883 +/- 1,542 U/ml) and IL-6 (966 +/- 619 ng/ml) in the exercised group, sampled 2h after LPS challenge, were significantly lower than those in the control group (6,571 +/- 2,766 U/ml and 2,428 +/- 1,228 ng/ml, respectively). It is suggested that the exercise-induced inhibition of endotoxin shock may be caused, at least partly, by the decreased production of IL-1 and IL-6. PMID- 7474535 TI - The effects of mild, moderate, and severe exercise on intraocular pressure in glaucoma patients. AB - BACKGROUND: Previous studies have shown a reduction in intraocular pressure (IOP) from many means of exertion, ranging from walking to exhausting exercise in both normal and glaucoma subjects. The variability in their results may be due to several variables that can affect IOP significantly. PURPOSE: To investigate the effects of common means of exertion on IOP in normal and glaucoma subjects, after elimination of factors that can affect IOP. SUBJECTS AND METHODS: The effects of walking, jogging, and running fast until exhaustion on IOP were noted in seven normal and seven open-angle glaucoma subjects. Intraocular pressures were measured with the Goldmann applanation tonometer during and after exercise. RESULTS: The mean differences between before and after walking, jogging, and running exercises were -2.43 +/- 0.30, -3.85 +/- 0.55, and -4.0 +/- 0.37 mmHg in normal subjects. In glaucoma patients these differences were -7.72 +/- 1.25, 10.86 +/- 2.12, and -12.86 +/- 2.05 mmHg, respectively. After 5 min of walking and jogging, in all subjects, IOP decreased significantly (by 56 to 61% of total decreases). CONCLUSION: Regardless of the means of exertion, in every subject tested there was a drop in intraocular pressure. Glaucoma patients had a greater drop and longer duration of post-exercise recovery as compared to normal subjects. It would seem reasonable, at present, not to discourage patients who have glaucoma from light exercises such as walking; perhaps, on the contrary, it should be encouraged. PMID- 7474536 TI - The sweating responses of athletes trained on land and in water. AB - In order to examine whether different sweating responses of athletes trained on land and in water may be ascribed to changes in the central sudomotor mechanisms and/or those of the peripheral mechanisms of sweat glands, we measured the local sweating rate at the left forearm (mswf) and the left scapula (msws), the frequence of sweat expulsion (Fsw) and body temperatures (mean skin temperature and rectal temperature: Tre) in six runners and five soccer players (R group) and six swimmers (S group) during progressive thermal stress at rest (2 degrees C increase in ambient temperature every 15 min from 35 to 45 degrees C RH = 30 40%). Tre and heart rate at the end of experiment did not differ significantly between the groups (37.31 +/- 0.04 degrees C, 74.5 +/- 7.9 beats.min-1 in the S group and 37.27 +/- 0.07 degrees C, 71.1 +/- 9.0 beats.min-1 in the R group, respectively). The msws and mswf at any given mean body temperature (Tb) were greater in the S group than in the R group. Although the regression line showing the relationship between Fsw and Tb in the S group was shifted to the left of that in the R group, there was no significant difference in the slope of the lines. The msws-Fsw or mswf-Fsw regression line was not different between the two groups. These results indicate that the higher sweating rate in the S group may be ascribed to a difference in the centrally derived sudomotor neural activity, but not to that in the peripheral mechanisms of sweat gland activity. PMID- 7474537 TI - A real-time observation of X-ray diffraction from frog skeletal muscle during and after slow length changes. AB - X-ray diffraction patterns from a frog skeletal muscle were recorded in real time using an X-ray image intensifier and a CCD video camera with a time resolution of 1/60s. The muscle was stimulated and then released or stretched by 6% at a constant speed. The equatorial (1,1) reflection decreased in intensity during stretches, suggesting a conformational change of cross-bridges. The integrated intensity of the 3rd meridional reflection from the thick filament decreased during both releases and stretches. The decrease was larger in stretches than in releases and at higher velocities. The intensity change correlated well with the change in tension, showing that the axial mass distribution of cross-bridges becomes wider as the tension increases. The axial Bragg spacing of the 3rd meridional reflection tended to increase in stretches and decrease in releases by less than 0.1%, possibly reflecting the elasticity in the thick filament backbone. The only significant effect of a length change remaining at the end of the stimulation was the broadening of the 3rd meridional reflection, suggesting non-uniformity of sarcomere lengths or axial misalignment of thick filaments as causes of enhancement and deficit of tension. PMID- 7474538 TI - Histamine H2 receptor-mediated endothelium-dependent relaxation in canine spinal artery. AB - The mechanisms of histamine-induced relaxation were investigated in isolated canine spinal branches of intercostal artery (SBICA). Histamine (10(-7) to 10( 4)M) and dimaprit (3 x 10(-6) to 3 x 10(-4)M) produced concentration-dependent relaxation in the SBICA which had been contracted by 5 x 10(-6)M norepinephrine, whereas 2-pyridylethylamine caused only a very small relaxation. The histamine- and dimaprit-induced relaxation were inhibited by famotidine but not by diphenhydramine. In the SBICA without endothelium, histamine elicited very little relaxation. Aspirin (5 x 10(-5)M) and AA 861 (10(-5)M) did not affect the histamine-induced relaxation. Treatment with L-NMMA (3 x 10(-5)M) or methylene blue (10(-5)M) significantly suppressed the acetylcholine-induced relaxation of the SBICA but not the histamine-induced one. These results suggest that histamine produces relaxation in the isolated canine SBICA through stimulation of H2 receptors on the endothelium. The relaxing mediator(s) released by histamine seems to be neither endogenous prostaglandins, lipoxygenase products, nor nitric oxide. PMID- 7474539 TI - Ca2+ increase and pH decrease induced by hypochlorous acid in single quiescent myocytes isolated from rat ventricles. AB - Cytosolic free calcium concentration ([Ca2+]i) and intracellular pH (pHi) were simultaneously measured with a fluorescence technique using fura-2 and BCECF. Hypochlorous acid (HOC1) increased [Ca2+]i and decreased pHi of single quiescent myocytes isolated from rat ventricles. The HOC1-induced changes in [Ca2+]i and pHi (delta [Ca2+]i and delta pHi) were 129 +/- 18nM and 0.18 +/- 0.02 (mean +/- SD), respectively, with 200 microM HOC1. A positive linear correlation was obtained between delta [Ca2+]i and delta pHi under various extracellular Ca2+, pH, and Na+ conditions. Chelation of extracellular Ca2+ depressed only one-fourth of delta [Ca2+]i, and Ca2+ antagonists (verapamil and nifedipine) did not reduce delta [Ca2+]i, indicating that delta [Ca2+]i originates mainly from intracellular Ca2+ stores. A disulfide-reducing reagent, 1,4-dithiothreitol (DTT), recovered the increased [Ca2+]i and decreased pHi to the control levels. The simultaneous changes in [Ca2+]i and pHi induced by HOC1 and the simultaneous restoration of the normal [Ca2+]i and pHi in cardiac myocytes. PMID- 7474540 TI - In vitro [U-14C]glucose utilization by tissues of weanling rats with lateral hypothalamic area lesions one month after lesion production. AB - The role of the lateral hypothalamic area (LHA) in intermediary metabolism was investigated by quantitation of [U-14C]glucose oxidation to 14CO2 and 14C incorporation into the glycogen and lipid fraction of the liver, epididymal fat pad, and diaphragm. Weanling male Sprague-Dawley rats received bilateral electrolytic lesions in the LHA (LHAL rats). Sham operated rats were either fed ad libitum (CON-ADLIB) or pair-gained to the LHAL rats (CON-PG). The experiment was terminated 1 month after lesion production. LHAL rats were significantly (SIG) lighter and shorter and ate less than CON-ADLIB; LHAL rats were also SIG shorter than CON-PG, pointing to a food intake-independent lesion effect. Both LHAL and CON-PG rats had SIG less percent carcass fat than CON-ADLIB, but there was no SIG difference between LHAL and CON-PG rats. Also, LHAL rats had a SIG higher percentage of carcass protein than both CON-ADLIB and CON-PG. Furthermore, LHAL rats incorporated SIG less glucose into liver glycogen than CON-ADLIB but SIG more into CON-PG, whereas CON-PG rats incorporated SIG less into liver glycogen than CON-ADLIB, again suggesting a food intake-independent effect. There was no difference among the groups in glucose oxidation and incorporation into lipids and glycogen in both diaphragm and epididymal fat pads and liver total lipid. However, livers of CON-PG metabolized SIG more [U-14C]glucose to CO2 than did livers of CON-ADLIB, suggesting a food intake-dependent effect. There was no difference between LHAL and CON-PG rats in this parameter.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7474541 TI - Effects of fasting and refeeding on some metabolic characteristics of rat brown adipose tissue. AB - To investigate the effects of fasting and refeeding on rat interscapular brown adipose tissue (BAT), we determined in vitro oxygen consumption, in vitro lipolysis, DNA content and composition of fatty acid (FA). Fasting decreased the weights of body and BAT, and the DNA content of BAT. Fasting reduced both basal oxygen consumption and basal lipolysis of BAT. Fasting also suppressed the noradrenaline- and glucagon-induced increases in both oxygen consumption and lipolysis of BAT. In both triglyceride and phospholipid FA of BAT the extent of unsaturation was increased by fasting. Refeeding restored all of these parameters to the control level. These observations indicate that fasting reduces both oxygen consumption and lipolysis of BAT, resulting in reduced thermogenesis, while fasting could stimulate BAT in certain phases of its function as observed in FA composition. PMID- 7474542 TI - Effect of a training session of endurance running on anserine and carnosine contents in fast and slow muscles of young rats. AB - The effect of a training session of endurance running on anserine and carnosine contents in extensor digitorum longus (EDL) and soleus muscles (SOL) of rats was studied during aging from 5 to 20 weeks. During aging from 5 to 14 weeks, the endurance training suppressed the developmental increases of body, EDL, and SOL weights in a similar way and of the anserine and carnosine contents in the two types of muscles to a greater extent than their weights. This indicates that the suppression of the developmental increase of the anserine and carnosine contents in the muscles by the training is not only due to their slow growth, but also to some, probably adaptational change of the muscle to the training. The suppressing effect of the training during the subsequent aging appeared to be equal to or even weaker than the before in these muscles. PMID- 7474543 TI - Inhibitory effect of light of different wavelengths on the fall of core temperature during the nighttime. AB - Nocturnal core temperature fall was significantly inhibited by green, blue, and red light exposure with 1,000 lx from 21:00 h to 02:00 h. The core temperature in red became identical from that in control during the following sleep period, but not in green and blue. These findings are discussed in terms of urinary melatonin behavior. PMID- 7474544 TI - GTP-binding protein-mediated production of superoxide anion in rabbit gastric parietal cells. AB - We found that guanosine-5'-O-(3-thiotriphosphate) (GTP gamma S) increased superoxide production in the suspension rich in rabbit gastric parietal cells that were permeabilized by streptolysin O. In this experiment, oxidation of non fluorescent dihydrorhodamine 123 into the fluorescent rhodamine 123 by oxygen radicals was measured. Results support that GTP gamma S closes a housekeeping C1- channel via intracellular production of superoxide anion in the parietal cell. PMID- 7474545 TI - A simple construction of dual-slit red cell velocimeter. AB - A simple method of building a dual-slit red cell velocimeter is introduced. The basis of this technique is to utilize the body of a 35-mm photomicrography camera as the photometric device, exactly centering a pair of highly sensitive photodiodes on the film plane of the camera. By this arrangement, the microvascular image can be accurately focused on the sensing windows of the photodiodes. Because this velocimeter is not based on videophotometry, it was possible to measure red cell velocity up to 30 mm/s in the benchmark test. The maximum red cell velocity actually measured in the rat mesenteric microvessels (6.0 to 48 microns in diam) was 9 mm/s. PMID- 7474546 TI - Two different endothelin B receptor subtypes mediate contraction of the rabbit saphenous vein. AB - To study endothelin receptor subtypes that mediate venous smooth muscle contraction, effects of some endothelin receptor agonists and antagonists on the rabbit lateral saphenous vein were examined and compared with those on the saphenous artery. In the artery, endothelin (ET)-1 elicited concentration dependent contractions, while selective ETB-receptor agonists, IRL1620 (Suc [Glu9,Ala11,15]ET-1(8-21)) and sarafotoxin 6c (S6c) had almost no effect. The ET 1-induced responses shifted in parallel to the right by BQ-123 (cyclo (-D-Trp-D Asp-Pro-D-Val-Leu-)), an ETA-receptor antagonist, or PD142893 (Ac-D-Dip-Leu-Asp Ile-Ile-Trp), an ETA/ETB-receptor antagonist, indicating the involvement of the ETA receptor in this response. In the saphenous vein, not only ET-1 and ET-3, but also ETB-receptor agonists, IRL1620, S6c and [Glu9]sarafotoxin 6b ([Glu9]S6b), produced concentration-dependent, BQ-123-insensitive contractions. PD142893 did not affect the ET-1-induced contraction, but it shifted greatly the IRL1620 induced concentration-response curve in parallel to the right. The major components of ET-3-, S6c- and [Glu9]S6b-induced contractions were resistant to PD142893. These results indicate that two different vasoconstrictive ETB-receptor subtypes, ETB1 (sensitive to IRL1620 and PD142893) and ETB2 (insensitive to IRL1620 and PD142893), are located on the smooth muscle of the saphenous vein. PMID- 7474547 TI - Bronchodilator and cardiovascular effects of NKH477, a novel water-soluble forskolin derivative, in guinea pigs. AB - The bronchodilator and cardiovascular effects of NKH477 (6-(3 dimethylaminopropionyl)forskolin hydrochloride) were evaluated. In anesthetized guinea pigs, i.v. bolus injections of NKH477 (1-100 micrograms/kg) inhibited the bronchoconstriction induced by inhaled leukotriene D4, increased the heart rate (HR) and decreased the diastolic arterial blood pressure (DBP) in a dose dependent manner. The bronchodilator effect of NKH477 was 1500 times more potent than that of aminophylline and 17 times less potent than that of isoproterenol. The selectivity of NKH477 for bronchodilation vs an increase in HR was 15 times higher than that of isoproterenol and similar to that of aminophylline; and vs a decrease in DBP, the selectivity was 4 times higher than that of aminophylline and similar to that of isoproterenol. I.v. infusion of NKH477 (0.1-3 micrograms/kg/min) for 2 hr dose-dependently inhibited the bronchoconstriction induced by i.v. histamine. Isoproterenol (0.1 microgram/kg/min, i.v.) enhanced the bronchoconstriction after termination of the infusion, whereas NKH477 did not. In conscious guinea pigs, inhalation of NKH477 (0.1-5 mg/ml) concentration dependently inhibited the bronchoconstriction induced by inhaled histamine, and a high concentration of NKH477 (35.4 mg/ml) increased the HR. The bronchodilator effect of inhaled NKH477 was 15 times less potent than that of isoproterenol. The selectivity of inhaled NKH477 was similar to that of isoproterenol. These results indicate that NKH477 may be useful as a bronchodilator. PMID- 7474549 TI - Effect of 5,6,7,8-tetrahydroneopterin on the bovine endothelial cell injury induced by cumene hydroperoxide. AB - Neopterin is an 2-amino-4-hydroxypteridine derivative and a precursor of biopterin, which is derived from guanosine triphosphate. Previously, we have reported that 5,6,7,8-tetrahydroneopterin (NPH4), a reduced form of neopterin, possesses an antioxidant activity in various systems. In this study, we investigated the activity in more detailed manner and discussed the possible applications of this antioxidant. Analysis by electron spin resonance spectrometry indicated that NPH4 scavenged superoxide anion radicals and hydroxyl radicals as well. Moreover, NPH4 protected the rat brain homogenate from autoxidation. Next, we examined the effect of NPH4 on the cell injury induced by cumene hydroperoxide (CHP) in cultured bovine artery endothelial cells. The activity of lactate dehydrogenase, a marker enzyme of cell injury, was elevated by CHP in a dose-dependent manner, and this elevation was dose-dependently suppressed by NPH4. The elevation of lipid peroxide content was also inhibited by NPH4 in the same fashion. These data suggest that NPH4 would be effective against various diseases whose pathogenesis is active oxygen-related. PMID- 7474548 TI - Alteration of bone status with ascorbic acid deficiency in ODS (osteogenic disorder Shionogi) rats. AB - Rats with hereditary defects in ascorbic acid (AsA) synthesis (ODS rats) subjected to AsA-deficiency for 3 weeks showed reductions of plasma alkaline phosphatase and dry and ash weights of the tibia, but no body weight alteration. In accordance with the chemical changes, bone loss and decrease of bone formation by AsA deficiency but not by malnutrition were observed in contact microradiographs of the tibia and by a tetracycline double labeling technique, respectively. The mechanical properties of femora measured by a three point bending procedure were also altered by AsA deficiency for 3 weeks and showed decreases of 59% in toughness, 32% in strength, 32% in ductility and 22% in stiffness. The biomechanical changes by AsA deficiency were greater than the chemical changes in bone, indicating the usefulness of measuring mechanical properties as a sensitive method for the evaluation of the bone status. The second moment of the area of the femur was not changed by AsA deficiency. These results suggest that AsA deficiency in ODS rats causes marked bone loss and reduction in bone formation, which is accompanied by a greater reduction in biomechanics of the femur without causing macroarchitectural changes. PMID- 7474550 TI - Inhibitory action of indomethacin on neutrophil infiltration in monosodium urate induced pleurisy in rats. AB - The effects of indomethacin on the production of cytokines at inflammatory sites were investigated in the monosodium urate (MSU) pleurisy model characterized by both cellular influx and edema. Indomethacin (10 mg/kg) orally administered 0.5 hr prior to MSU injection into the pleural cavity significantly inhibited MSU induced neutrophil accumulation in the cavity. In addition, the drug slightly enhanced the level of MSU-induced tumor necrosis factor production without affecting interleukin-1 production. Furthermore, indomethacin inhibited both the levels of MSU-induced rat cytokine-induced neutrophil chemoattractant (CINC/gro) and interleukin-6 (IL-6) production by 78.3% at 3 hr and 45.8% at 4 hr post injection, respectively. Although intrapleural injection of CINC/gro induced neutrophil infiltration in a dose-dependent manner, IL-6 did not affect the action of CINC/gro on neutrophil influx. These findings suggest that the inhibitory action of indomethacin on neutrophil infiltration is, at least, partly mediated by a decrease in the MSU-induced CINC/gro content in this model. PMID- 7474551 TI - Non-selective effects of amiloride and its analogues on ion transport systems and their cytotoxicities in cardiac myocytes. AB - The effects of amiloride and its analogues (3',4'-dichlorobenzamil (DCB), 2',4' dimethylbenzamil (DMB), 5-(N-ethyl-N-isopropyl)amiloride (EIPA) and 5-(N-methyl-N isobutyl)amiloride (MIBA)) on cardiac ion transporters (Na+/Ca2+ exchanger, Na+/H+ exchanger, Na+ pump and Ca2+ pump) and their cytotoxicities were tested in cardiac myocytes. All the tested compounds showed concentration-dependent inhibitory effects on the ion transporters studied in canine cardiac sarcolemmal vesicles. The concentrations (microM) of amiloride, DCB, DMB, EIPA and MIBA required to produce 50% inhibition were > 1000, 19, 10, 83 and 84, respectively, for the Na+/Ca2+ exchanger; 130, 73, 63, 16 and 14 for the Na+/H+ exchanger; > 1000, 72, > 300, > 300 and > 300 for the Na+ pump; and > 1000, 37, 93, 90 and 70 for the Ca2+ pump, respectively. Furthermore, these agents induced cell death in isolated rat cardiac myocytes and the 50% lethal concentrations (microM) were > 1000, 9.2, 30, 16 and 17, respectively. These findings demonstrate that amiloride and its analogues have non-selective inhibitory effects on cardiac ion transporters and cytotoxicity in cardiomyocytes. When these drugs are employed as experimental tools to investigate the involvement of ion transporters in cell functions, the results must be interpreted with caution. PMID- 7474552 TI - Healing-promoting action of the zinc-cimetidine complex on acetic acid-induced gastric ulcers in rats. AB - We investigated the effect of the zinc-cimetidine complex on the healing of acetic acid-induced gastric ulcers in rats. When the effects of test drugs were assessed on the 15th day after acetic acid injection, the zinc-cimetidine complex at oral doses of 15.0 (11.4 mg as cimetidine), 30.0 and 60.0 mg/kg twice daily promoted the ulcer healing in a dose-dependent manner. Cimetidine was effective at oral doses of over 45.4 mg/kg twice daily. ZnCl2 was ineffective on all ulcer parameters. The effect of the combination of cimetidine and ZnCl2 was similar to that of cimetidine alone. The zinc-cimetidine complex had already inhibited the increase in thiobarbituric acid reactants in the ulcerated region before the ulcer-healing effect of this compound was recognized. A single oral administration of the complex at 15 and 30 mg/kg to normal rats was ineffective in inhibiting acid secretion and in increasing serum gastrin levels, although cimetidine was markedly effective on both parameters. These results indicate that the zinc-cimetidine complex at about 1/4 the dose of cimetidine was as effective as cimetidine when the ulcer-healing effects of both compounds were compared with the same dose of cimetidine. In addition, the ulcer-healing effect of this complex may be due, at least in part, to the inhibition of lipid peroxidation but not due to the inhibition of acid secretion or the trophic effect of gastrin. PMID- 7474553 TI - The uricosuric effect in rats of E5050, a new derivative of ethanolamine, involves inhibition of the tubular postsecretory reabsorption of urate. AB - N-[3-[4'-(2",6"-Dimethylheptyl)phenyl]butanoyl]ethanolamine (E5050), a newly synthesized compound, was shown recently to induce uricosuria in humans via inhibition of the postsecretory reabsorption of urate. We examined the effects of this compound on urate excretion in rats loaded with oxonate and compared these effects with those of the uricosuric drugs trichlormethiazide and probenecid. When administered i.p., E5050 (0.3-15 mg/kg) increased the urinary excretion rate of urate and the ratio of urate clearance to inulin clearance in a dose-dependent manner, while the urine volume increased only slightly, and the glomerular filtration rate and plasma urate level were not changed. No paradoxical effect on urate excretion was observed. In contrast, trichlormethiazide and probenecid had a biphasic effect on urate excretion. In a pyrazinoic acid suppression test, the uricosuric effect of E5050 was completely inhibited by pretreatment with pyrazinoic acid. In a phenolsulfonphthalein (PSP) test, E5050 did not affect urinary PSP excretion, while probenecid strongly decreased such excretion. Thus, E5050 also appears to be uricosuric in rats. PMID- 7474554 TI - Smooth muscle relaxing and hypotensive activities of synthetic calciseptine and the homologous snake venom peptide FS2. AB - The biological activities of synthetic calciseptine and FS2, a homologous peptide from snake venom, were determined using in vitro and in vivo preparations. Calciseptine and FS2 produced dose-dependent relaxation in pre-constricted rat aorta, pulmonary artery and trachea. The onset and duration pattern of these relaxing effects were similar to those caused by nifedipine, an L-type Ca2+ channel blocker. Calciseptine relaxed the contraction of rat aorta provoked by an L-type channel agonist, Bay K 8644. This relaxation was not affected by NG-nitro L-arginine, indomethacin or propranolol. Calciseptine and FS2 inhibited the contraction caused by acetylcholine in guinea pig ileal longitudinal muscle. In case of in vivo study using anesthetized rats, calciseptine, FS2 and nifedipine showed depressor effects. The hypotensive effects of the two peptides were more potent and sustained than that of nifedipine. These findings show that both synthetic calciseptine and FS2 have similar biological activities like nifedipine, an L-type Ca2+ channel blocker. In addition, these two peptides with large molecular weights may be unique and useful tools for studying the Ca2+ channel. PMID- 7474555 TI - Potentiation of excito-toxicity by glutamate uptake inhibitor rather than glutamine synthetase inhibitor. AB - The neuroprotective functions of glia cells in the presence of excessive amounts of extracellular glutamate (Glu) were examined using glia-rich and glia-poor cultured cerebellar granule cells that contained the same number of neurons. In order to focus on the metabolic enzyme glutamine synthetase (GS) and the uptake system in glia cells, selective inhibitors such as L-methionine sulfoximine (MSO) and 4-acetamido-4'-isothiocyanostilbene-2,2'-disulfonic acid (SITS) were used as pharmacological tools. The increased amount of lactate dehydrogenase (LDH) leakage induced by 50 microM Glu and SITS was equivalent to that of 1 mM Glu. However, the simultaneous treatment with 50 microM Glu and 5 microM MSO did not increase the LDH leakage. The larger quantities of extracellular Glu were sustained in both glia-rich and glia-poor cultures. After the administration of Glu and MSO, however, the larger quantities of Glu were not sustained. Taking these results into consideration, the Glu uptake system in glia cells seems to be more important than the Glu metabolic enzyme system in the regulation of neuronal protection from Glu toxicity. PMID- 7474556 TI - Antioxidative action of the beta-adrenoceptor antagonist bopindolol and its metabolite 18-502. AB - The antioxidative effects of beta-adrenoceptor antagonists and related compounds were investigated. Among the beta-adrenoceptor antagonists, the agents with a potent membrane-stabilizing activity such as bopindolol and propranolol strongly inhibited the hydrogen peroxide (H2O2)-induced lipid peroxidation of liver microsomes. Fifty percent inhibition concentration values for the lipid peroxidation of bopindolol, 18-502 (metabolite of bopindolol) and propranolol were calculated to be 1.8 microM, 10 microM and 2.3 microM, respectively. The same potency order of the agents for the inhibition of lipid peroxidation was observed in rat heart homogenates. Furthermore, cytochrome P-450-catalyzing lipid peroxidation in microsomes and H2O2-induced lipid peroxidation in coronary arteries or cardiac muscles of pigs were also inhibited by bopindolol, whereas propranolol was less effective. Bopindolol and 18-502, but not propranolol, scavenged a stable free radical 1,1-diphenyl-2-picrylhydrazyl. Thus it was concluded that bopindolol that has membrane-stabilizing and radical scavenging activities is a more potent antioxidant than propranolol and may produce a beneficial effect for the treatment of ischemic cardiac diseases. PMID- 7474557 TI - Role of endothelium in oscillatory contractile responses to various receptor agonists in isolated small mesenteric and epicardial coronary arteries. AB - Employing the isometric tension recording method, we studied the role of endothelium or endothelium-derived relaxing factor (EDRF) in the generation of rhythmic oscillations observed in contractile responses to various receptor agonists in isolated rabbit small mesenteric and epicardial coronary arteries. Norepinephrine (NE, 0.1-10 microM) generated oscillatory contraction in endothelium-intact strips from the mesenteric arteries. Similarly, acetylcholine (ACh, 10 microM), histamine (10 microM) and serotonin (10 microM) generated oscillatory contraction in endothelium-intact strips from the coronary arteries. These agonist-induced oscillations in both arteries were consistently eliminated by either endothelial denudation or EDRF pathway inhibitors including NG-nitro L arginine (30 and 100 microM), oxyhemoglobin (3 and 10 microM) and methylene blue (3 and 10 microM). In contrast, EDRF releasers such as ACh or A23187 augmented the oscillations in the endothelium-intact strips. SNP (0.03-30 microM) failed to generate oscillations in NE (10 microM)-preconstricted endothelium-denuded strips from the mesenteric arteries. In conclusion, these agonist-induced oscillations are probably mediated through EDRF. The inability of SNP to generate oscillations suggests the obligatory role of the endothelium in generation of the oscillations. The oscillatory release of EDRF by endothelial cells may be responsible for generation of the oscillations. PMID- 7474558 TI - OPC-21268 antagonizes arginine vasopressin-induced vasoconstrictor response in the spinally-anesthetized dog. AB - We studied the antagonistic actions of OPC-21268 (1-(1-[4-(3-acetylaminopropoxy) benzoyl]-4-piperidyl)-3,4-dihydro- 2(1H)-quinolinone) on the arginine vasopressin (AVP)-induced vasoconstrictor response in the spinally-anesthetized dog. OPC 21268 at doses of 0.3, 1.0 and 3.0 mg/kg, i.v. produced a rightward parallel shift of the dose-response curves for AVP in a dose-dependent manner. The doses of OPC-21268 were similar to those that inhibited the AVP-induced vasoconstrictor response in the rat. This observation suggests that OPC-21268 acts as a V1-AVP receptor antagonist in peripheral resistance vessels in dogs as well as in rats. PMID- 7474559 TI - The effects of thiamine and oxythiamine on the survival of cultured brain neurons. AB - The effects of treatment with thiamine (Vitamin B1) alone or together with its antagonist oxythiamine on the survival of brain neurons in primary culture were investigated. Treatment with thiamine significantly promoted the survival of hippocampal neurons in high cell density culture, but had no effects on the neuronal survival in low cell density culture. In addition, the survival promoting activity exerted by thiamine was remarkably decreased by the co application of oxythiamine, although oxythiamine used alone revealed neither a trophic nor toxic effect on the neurons of examined brain regions. The neurotrophic function of thiamine may be due to its coenzymatic role in a biochemical reaction and/or its specific function on neurotransmission and nerve conduction. PMID- 7474560 TI - Inhibitory effect of KBT-3022, a new anti-platelet agent, on infiltration of polymorphonuclear leukocytes induced by leukotriene B4 or formyl-methionyl-leucyl phenylalanine in mice. AB - We devised a method for evaluating polymorphonuclear leukocyte (PMN) infiltration in vivo employing an air bleb technique combined with measurement of myeloperoxidase (MPO) activity, and the effects of some anti-platelet agents were evaluated. KBT-3022 (ethyl 2-[4,5-bis(4-methoxyphenyl)thiazol-2-yl]pyrrol-1 ylacetate) and cilostazol inhibited the increase in MPO activity in the connective tissue around the air bleb induced by leukotriene B4 (LTB4) and formyl methionyl-leucyl-phenylalanine (fMLP). Indomethacin inhibited only the fMLP induced increase in MPO activity, but ticlopidine hydrochloride and acetylsalicylic acid had no effect. Histologic observation confirmed the inhibition of PMN infiltration by KBT-3022. These results indicate that KBT-3022 may be a potent inhibitor of both LTB4- and fMLP-induced infiltration of PMNs. PMID- 7474561 TI - [Respiratory disorders during sleep in Duchenne muscular dystrophy]. AB - Respiratory disorders during sleep were studied in 42 patients with Duchenne muscular dystrophy (DMD) (mean age 18.4 years). Chest and abdominal movement, nasal airflow, snoring sounds, eye movement, and oxygen saturation were monitored during sleep. Three patterns of disorders were found: obstructive apnea, central apnea, and paradoxical respiration without upper airway obstruction (non obstructive paradoxical respiration). Of these three patterns, obstructive apnea was the most common. Hypertrophy of the tongue and collapsibility of the upper airway seemed to be responsible for the obstructive apnea in these patients. The relationships between PaCO2 while breathing room air and the various indices of respiratory disorders were studied. The index of central apnea differed significantly between patients in whom PaCO2 was less than 50 Torr (early disease, n = 22) and those in whom PaCO2 was greater than or equal to 50 Torr (advanced disease, n = 20), but the indices of obstructive apnea and non obstructive paradoxical respiration did not differ between those two groups. In conclusion, sleep disorders were common in patients with DMD, and the most common was obstructive apnea. In the patients with advanced DMD, blood gas analysis showed hypercapnia, and the index of central sleep apnea was high, probably because of respiratory muscle weakness or abnormalities in the respiratory center. PMID- 7474562 TI - [Small airway changes in acute interstitial pneumonia and acute exacerbation of chronic interstitial pneumonia]. AB - To study pathologic changes in small airways in acute interstitial pneumonia (AI), changes in the terminal or respiratory bronchioles in one patient with AI and in three patients with acute exacerbations of chronic interstitial pneumonia (grouped together as the AI group) were compared to those in five patients with chronic idiopathic interstitial pneumonia (IIP) without acute exacerbation. Specimens obtained during autopsy were examined by light microscopy. In the AI group, mononuclear cells had infiltrated the lumen in 47 (23.7%) of the 198 bronchioles examined, and granulation tissue was found in 101 (51.0%). In the IIP group, no changes were found in 37 (97.4%) of the 38 bronchioles examined. Therefore, mononuclear cell infiltration and granulation tissue are common in the bronchioles of patients with acute interstitial pneumonia, but not in those of patients with chronic interstitial pneumonia without acute exacerbation. PMID- 7474563 TI - [Primary pulmonary alveolar proteinosis--clinical observation of 68 patients in Japan]. AB - The clinical features of 68 Japanese patients (53 men and 15 women; mean age 44 years) with primary pulmonary alveolar proteinosis were reviewed. Pulmonary alveolar proteinosis was diagnosed from histologic findings after open lung biopsy (n = 7) or transbronchial lung biopsy (n = 61). Major symptoms were a dry cough (24.2%) and dyspnea or shortness of breath on exertion (51.5%), but one third of the patients were asymptomatic. Crackles were audible in 30% of the patients, but clubbing (6%) and cyanosis (4%) were rare. Ten patients had been occupationally exposed to dust. Slightly less than half (46%) of the patients first presented with symptoms, and the remainder (54%) first presented with abnormal pulmonary infiltrates seen on chest roentgenograms taken during general health examinations. Many patients had abnormally high levels of LDH and CEA in serum (62% and 63%, respectively). Restrictive pulmonary dysfunction (%VC < 80%) was seen in 31% of the patients, an abnormally low DLco (%DLco < 70%) was seen in 62%,m and hypoxemia (PaO2 < 80 mmHg) was seen in 67%. Arterial blood gas tension was closely correlated with the severity of disease in these patients. Chest roentgenograms usually showed bilateral symmetric alveolar infiltrates, mainly distributed from hilar areas toward the pleura, but on CT scans many of the shadows were mixed with alveolar and interstitial infiltrates of various extent along the pulmonary arteries and bronchi. There was no apparent relation between chest roentgenographic findings and chest CT findings in these patients. Neither the extension nor other characteristics of shadows in the chest roentgenograms and chest CT scans were closely related to symptoms, laboratory data, or pulmonary function in these patients. Symptoms were alleviated and chest roentgenographic findings improved in 82% of the 51 patients who underwent therapeutic bronchoalveolar lavage, and in 94% of the 17 patients who did not undergo that procedure. In patients who underwent therapeutic bronchoalveolar lavage and also in those who recovered spontaneously, both diffusing capacity and blood gas values improved significantly. When compared to the patients who did not undergo therapeutic bronchoalveolar lavage, significantly more of those who did undergo that procedure has initial PaO2 values below 60 mmHg, and fewer of them had values greater than 80 mmHg. Thus, a PaO2 below 60 mmHg may be an indication for therapeutic bronchoalveolar lavage in patients with this disease. During the follow-up period (mean 5 years, range 2 months to 23 years), four patients had pneumothorax and none died of pulmonary alveolar proteinosis. PMID- 7474564 TI - [Thyroid hormones in patients with acute exacerbations of pneumoconiosis]. AB - Thyroid hormone levels in serum were measured in 34 patients with acute exacerbations of pneumoconiosis. Levels of free T3 were below the normal range in 26 patients, and 12 of those patients (46.2%) died. Levels of free T3 were within the normal range in 8 patients, and only of those patients died. Having a T3 level within the normal range was associated with a good outcome. Levels of free T3 measured during acute exacerbations were significantly lower than those measured when the disease was clinically stable. Patients who died tended to have lower levels of free T3 than did those who survived. These results suggest that levels of free T3 in serum samples taken during an acute exacerbation can helpful in predicting the outcome of pneumoconiosis. PMID- 7474565 TI - [Serum levels of soluble receptors for tumor necrosis factor in sarcoidosis]. AB - Tumor necrosis factor alpha (TNF-alpha) has been shown to play an important role in granulomatous diseases, including sarcoidosis. TNF-alpha starts affecting cell function by binding to specific, high-affinity receptors on the cell surface, and two types of TNF-alpha receptors have been identified. Recently, soluble forms derived these cell surface receptors (sTNF-R type I and type II) have been shown to exist and have been investigated in several diseases. The levels of sTNF-R type I and type II in serum from patients with sarcoidosis were measured, and the clinical significance of sTNF-R was evaluated. The levels of both sTNF-R type I and type II were significantly higher in serum from patients with sarcoidosis (n = 36) than in serum from control subjects (n = 15): type I, 1.93 +/- 1.28 ng/ml vs 1.31 +/- 0.40 ng/ml, p < 0.01; type II, 3.48 +/- 2.60 ng/ml vs 1.56 +/- 0.35 ng/ml, p < 0.001. The levels of these receptors were significantly higher in patients with active sarcoidosis than in those with inactive sarcoidosis: type I, 2.43 +/- 1.83 ng/ml vs 1.57 +/- 0.45 ng/ml, p < 0.05; type 4.71 +/- 2.24 ng/ml vs 2.25 +/- 0.77 ng/ml, p < 0.01. The levels of sTNF-R type I and type II correlated significantly with the level of ACE, r = 0.70, p < 0.01; and r = 0.55, p < 0.05, respectively. We conclude that measurement of the levels of both types of sTNF-R may be useful in the evaluation of disease activity in sarcoidosis. PMID- 7474566 TI - [Effects of a beta 2-agonist, sodium cromoglycate, and an anticholinergic agent on hyperventilation-induced bronchoconstriction in sensitized rabbits]. AB - Hyperventilation can induce bronchoconstriction in ovalbumin-sensitized rabbits. To investigate the roles of the beta-receptor parasympathetic nervous system and of chemical mediators in hyperventilation-induced bronchoconstriction (HIB), the effects of a beta 2-agonist, of sodium cromoglycate, and of an anticholinergic agent on HIB were studied. Rabbits were divided to four groups and treated as follows. Group 1: Control (n = 7, 0.9% saline); Group 2: Procaterol (n = 4, 50 micrograms/l); Group 3: Sodium cromoglycate (n = 4, 10 mg/ml); and Group 4: Ipratropium bromide (n = 6, 1 mg/ml). Each drug was inhaled for 1 min via an ultrasonic nebulizer. Then, for the eucapnic hyperventilation challenge, sensitized rabbits were mechanically hyperventilated for 15 min (120 breaths/min, tidal volume = 7 ml/kg) with dry air containing 5% CO2 at room temperature. Total lung resistance and dynamic compliance were measured before, and 0, 5, 15, and 30 min after hyperventilation. The mean percent change in resistance measured 5 min after the hyperventilation was +49% in group 1, -6% in group 2, +23% in group 3, and +1% in group 4. The changes in groups 2 and 4 were significantly less than in group 1 (p < 0.05). In conclusion, HIB is mainly caused by bronchial smooth muscle constriction, and chemical mediators and the parasympathetic nervous system may play important roles in the development of HIB in sensitized rabbits. PMID- 7474568 TI - [Effect of adhesiveness of white blood cells on pulmonary vascular permeability and resistance]. AB - We studied pulmonary vascular injury evoked by mechanically activated white blood cells (WBCs). In isolated perfused rat lungs, changes in pulmonary vascular resistance were measured, and a gravimetric method was used to measure the pulmonary vascular filtration coefficient, which was taken as an index of pulmonary vascular injury. WBCs were activated by gentle agitation in a glass container for 10 sec. The filtration coefficient was measured under baseline conditions, and then inactive or activated WBCs were added to the perfusate. The perfusion was stopped for 90 minutes and then the lungs were reperfused. The expression of CD18 on the surface of WBCs was measured by flow cytometry. The filtration coefficient and the pulmonary vascular resistance in lungs exposed to activated WBCs were about 2.5 times and 3.3 times higher, respectively, than those in the lungs exposed to inactive WBCs. These results indicate increases in resistance and in pulmonary vascular permeability. The results of flow cytometry indicated that the mechanical agitation increased the expression of CD18 on the surface of WBCs. In conclusion, WBCs in which adhesiveness has been increased can induce pulmonary vascular injury and can increase pulmonary vascular resistance. PMID- 7474569 TI - [Squamous cell carcinoma of the lung with high CT number, but without calcification]. AB - Squamous cell carcinoma of the lung was diagnosed in a 64-year-old man. Chest roentgenographic and tomographic examinations did not show areas of high-density in the tumor, but chest CT scan showed diffuse, scattered, high-density nodules in the tumor. Histologic examination did not show calcification, and calcium staining (Kossa method) was negative. Examination of the thin-slice CT, the soft X-ray film, and the resected slice of the lung indicated that the high-density nodules seen in the tumor on the chest CT scan corresponded to markedly fibrotic lesions with severe anthracosis. This case shows that high-density areas on a CT scan may reflect non-calcified lesions. PMID- 7474570 TI - [Remission and relapse of acute eosinophilic pneumonia]. AB - A 46-year-old man presented with progressive dyspnea of acute onset, nonproductive coughing, and a high fever. He had been in his usual good health until symptoms began on the previous day. The chest roentgenogram revealed Kerley A and B lines, perivascular cuffing, hilar haze, and bilateral pleural effusins. Body temperature was 37 degrees C and PaO2 was 42 Torr. All clinical and radiographic signs improved. On the 12th hospital day, dyspnea and diffuse infiltration shadows on the chest roentgenogram suddenly occurred again. Acute eosinophilic pneumonia was strongly suspected because of increasing peripheral eosinophilia, and from the results of histologic examination of a specimen obtained by transbronchial lung biopsy. Without steroid treatment, the patient's condition gradually improved. Most cases of eosinophilic pneumonia have been diagnosed as pulmonary infiltration with eosinophilia (PIE). However, this patient did not have PIE syndrome, but instead was given a diagnosis of acute eosinophilic pneumonia, which was first described in 1990. This case may help establish criteria for the diagnosis and steroid treatment of acute eosinophilic pneumonia. PMID- 7474567 TI - [Effects of prostaglandin E1 (PGE1) on pulmonary hypertension and lung vascular remodeling in a rat monocrotaline model of human pulmonary hypertension]. AB - Prostaglandin E1 (PGE1) has been shown to be a potent pulmonary vasodilator in humans and in many animals. The effects of PGE1 on the development of pulmonary hypertension and on pulmonary vascular remodeling were studied in a rat monocrotaline (MCT) model of human pulmonary hypertension. By 3 weeks after injection, MCT (80 mg/kg S.C.) had resulted in high values of mean pulmonary arterial pressure and of the ratio of right ventricular weight to left ventricle+septum weight (RV/LV+S). PGE1 inhibited the development of pulmonary hypertension (300 micrograms/kg) and right ventricular hypertrophy (300 and 100 micrograms/kg) induced by MCT. Three weeks after the injection, the media walls of pulmonary arteries in lungs from rats given MCT were significantly thicker than those from lungs of control rats. PGE1 (300, 100, and 30 micrograms/kg) resulted in significantly less of this morphologic change, in a dose-dependent manner. These results indicate that PGE1 inhibits the development of pulmonary hypertension associated with lung vascular thickening induced by MCT. PGE1 may be useful for the treatment of pulmonary hypertension in humans. PMID- 7474571 TI - [Puerperal secondary pulmonary hypertension in a patient with mixed connective tissue disease]. AB - A woman with mixed connective tissue disease (MCTD) developed pulmonary hypertension after delivery of a child, but had little evidence of parenchymal lung disease. This 29-year-old woman had been given a diagnosis of MCTD when she was 19 years old. She was admitted to our department two days after delivery of a child, because of dyspnea on exertion. Acute thromboembolism was suspected because of: (1) chest roentgenogram showing cardiomegaly and enlargement of the left main pulmonary artery, (2) a lung perfusion scan showing a segmental defect in the left S6 and S8 areas, (3) laboratory studies showing abnormally high WBC, LDH, FDP, and D-D dimer, and (4) arterial blood gas analyses showing mild hypoxemia and hypocapnia. Thrombolytic therapy with heparin and urokinase was begun, and was followed by a loop diurtic and anticoagulation with warfarin. One month after admission, cardiac enlargement and the A-aDO2 were found to have decreased. At that time, cardiac catheterization was done and revealed pulmonary hypertension (mean PA pressure: 45 mmHg) and low cardiac output with no detectable thrombosis in the left pulmonary artery. The patient was subsequently treated with a calcium antagonist and a prostacyclin derivative, and her condition was stable for 5 months. Then her exercise tolerance gradually decreased due to shortness of breath, and cardiomegaly gradually increased over the next 3 months. Eight months after delivery of the child, the patient died of right heart failure. In clinically stable patients with MCTD, delivery of a child may lead to pulmonary thromboembolism and pulmonary hypertension. PMID- 7474572 TI - [Pulmonary endometriosis with recurrent catamenial hemoptysis]. AB - A 32-year-old woman with a history of dilation and curettage for missed abortion at 31 years old had sudden hemoptysis during menstruation on May 25, 1993. She had catamenial hemoptysis again, and was admitted to our hospital on July 21, 1993. After admission, catamenial hemoptysis occurred, and a nodular shadow was noted in the right S4 area on chest roentgenogram and computed tomogram. A few days after menstruation, hemoptysis and the abnormal shadow on chest roentgenogram disappeared. Pulmonary endometriosis was diagnosed by clinical course, past history, and chest roentgenographic findings. During the next menstruation, hemoptysis and a nodular shadow in the right S4 area on chest roentgenogram recurred, so transbronchial lung biopsy and laparoscopy were done. TBLB specimens revealed macrophages with phagocytosed hemosiderin, and laparoscopy revealed pelvic endometriosis. She was treated with buserelin acetate. Catamenial hemoptysis and the nodular shadow on the chest roentgenogram disappeared after treatment. PMID- 7474573 TI - [Giant cell interstitial pneumonia in a metal grinder with an abnormally high level of serum CA19-9]. AB - Interstitial pneumonia and recurrent pneumothorax developed in a 48-year-old man who had worked as a metal grinder. He died of respiratory failure despite having received antibiotics and steroids, and despite having undergone pleural sclerosis therapy. Giant cell interstitial pneumonia was diagnosed; innumerable bizarre giant cells engulfing black granules were found within the alveoli. The results of high-energy dispersion X-ray microanalysis indicated that the patient had hard metal pneumoconiosis associated with tungsten in the black granules. When he was admitted to the hospital, his serum CA19-9 and SLEX concentrations were abnormally high (2600 and 200 ng/ml, respectively). Immunohistochemical analysis of lung tissue was done with anti-CA19-9 and SLEX antibodies. CA19-9 staining revealed strong bronchialization and squamous metaplasia in contrast to type II hyperplasia. SLEX staining showed strong type II hyperplasia. Further investigations will be needed to determine the mechanism of elevated tumor associated carbohydrate antigens in serum. PMID- 7474574 TI - [Sarcoidosis with hypercalcemia and increased 1 alpha, 25-dihydroxyvitamin D in lung tissue]. AB - A 60-year-old woman was admitted to our hospital complaining of nausea, thirst, and dyspnea on exertion. She had skin eruptions on her back and right knee. Her serum calcium level was 12.6 mg/dl. The chest roentgenogram showed diffuse nodular shadows in both lung fields. The level of angiotensin-converting enzyme in serum was markedly high, and epithelioid cell granulomas were found in the specimens obtained by skin biopsy and by transbronchial lung biopsy. Sarcoidosis was diagnosed. After administration of prednisolone, the serum calcium level decreased to within the normal range and the shadows on the chest roentgenogram resolved. Hypercalcemia complicated with sarcoidosis is caused by overproduction of 1 alpha,25-dihydroxyvitamin D in granulomas. The content of 1 alpha, 25 dihydroxyvitamin D in the transbronchial lung biopsy specimen from this patient was higher than that from a sarcoidosis patient whose serum calcium level was normal. A few case reports of sarcoidosis with hypercalcemia have been published in Japan. The association with HLA type and the possibility of overintake of dietary vitamin D are discussed in reference to hypercalcemia in this patient. PMID- 7474575 TI - [A case of large cell carcinoma of the lung arising from the inner surface of a pulmonary bulla and complicated by hematoma]. AB - A 64-year-old man with a history of smoking was admitted to our hospital, because he was noted to have a solitary mass lesion at the apex of the right lung on a chest roentgenogram. Eight months before admission, he had come to our hospital because of hemoptysis. At that time, however, no abnormal shadow was seen on his chest roentgemogram, except for multiple bullae at both apexes. Based on chest CT findings on admission, the tumor appeared to be a hematoma growing inside the bulla. The resected tumor was found to contain a large amount of coagulated blood in the bulla. Histopathological examination of the bulla revealed a proliferation of large atypical cells from the inner surface of the bulla toward the inner space. Thus, the diagnosis was large cell carcinoma within the wall of the pulmonary bulla, the inside of which was filled with hematoma. We believe that the hematoma in the bulla allowed us to make an early diagnosis, and thus to succeed in curing the patient. PMID- 7474577 TI - [Analysis of tracheostomy for patients after thoracic esophageal cancer resection]. AB - Tracheostomy is an available method for respiratory management, but a consensus has not been reached with regard to the significance of postoperative respiratory management. So, we analyzed postoperative tracheostomy patients of esophageal cancer and clarified its significance. We analyzed 26 patients from among 85 who underwent resection for thoracic esophageal cancer between April 1989 and December 1992. We divided them into 3 groups according to the indications of tracheostomy, group I: cases with postoperative pulmonary complications (n: 17), group II: cases of prophylaxis against pulmonary complications (n: 5) and group III: cases with dyspnea deriving from postoperative recurrent nerve palsy (n: 4). Tracheostomy was most often performed between 3 and 4 POD in group I, up to 1 POD in group II and from 5 POD to 7 POD in group III. Long-term respiratory support was needed in 13 cases in group I and 2 cases in group II. The rate of tracheostomy was higher in the case who had postoperative recurrent nerve palsy and received preoperative chemoradiotherapy. All cases excluding a case of MOF had a good clinical course after tracheostomy. We were able to clarify the significance of tracheostomy by dividing tracheostomy patients into 3 groups according to the indications for the procedure. It was concluded that tracheostomy is a very useful technique in the respiratory management of resected thoracic esophageal cancer patients. PMID- 7474576 TI - [Pulmonary malignant lymphoma associated with bilateral diffuse linear, reticular, and consolidated shadows]. AB - A 72-year-old asymptomatic man was admitted to our hospital with abnormal chest radiographic findings: bilateral diffuse linear, reticular, and consolidated shadows. Malignant lymphoma (diffuse small-cell B-cell lymphoma) was diagnosed histologically after an open lung biopsy and was confirmed by a genetic examination. The lymphoma cells had infiltrated widely in various regions of the lung, including the alveolar spaces and walls, the bronchial and vascular spaces, and the pleura. This diffusion of the lymphoma cells might have accounted for the chest X-ray findings. Malignant pulmonary lymphoma should be included in the differential diagnosis patients with diffuse linear, reticular, and consolidated shadows on chest X-ray films. PMID- 7474579 TI - [The long-term results of isolated aortic valve replacement with 19 mm prostheses]. AB - Twenty-five patients underwent isolated aortic valve replacement with 19 mm valves (8 cases with St. Jude Medical prostheses, 7 cases with Carpentier Edwards Pericardial prostheses and 10 cases with Bjork-Shiley prostheses). Postoperative echocardiography revealed data as follows. Peak pressure gradients (mmHg) were 40.4 +/- 14.4 (mean +/- S.D.) in the SJM group, 23.4 +/- 9.8 in the CEP group and 50.5 +/- 16.7 in the BS group. End-diastolic left ventricular dimension in the BS group and interventricular septum thickness in the CEP group was reduced significantly. All but one case were in NYHA functional class I or II, and clinical improvement was almost satisfactory in all groups. However, in the BS group, 4 cases showed peak pressure gradients more than 50 mmHg. We conclude that 19 mm aortic prostheses in this study can provide satisfactory results. However, as for mechanical valve, it may be undesirable to choose BS prosthesis. PMID- 7474580 TI - [An autopsy case of immature teratoma with choriocarcinoma in the mediastinum]. AB - The autopsy of a 28-year-old Japanese male patient, which revealed immature teratoma combined with choriocarcinoma in the mediastinum, is presented. The tumor, which was removed after chemotherapy, was localized in the superior anterior mediastinum. Chemotherapy was performed several times after operation. However, HCG increased again, and a roentgenogram revealed metastatic shadows in both lungs. The metastatic tumors in the lungs showed only the choriocarcinoma. PMID- 7474578 TI - [Supra-annular mitral valve replacement for a patient with prosthetic valve endocarditis]. AB - Supraanular mitral valve replacement with a 29 mm CarboMedics prosthetic valve was performed for a 58-year-old patient of prosthetic valve endocarditis. He has received mitral valve replacement 2 years ago, and was admitted because of high fever and severe heart failure diagnosed mitral regurgitation due to valve dehiscence. An annular abscess was observed and the new 29 mm CarboMedics prosthetic valve was sutured to the left atrial wall approximately 10 mm above the original annulus in order to secure the firm suturings for valve implantation and to left the infected annulus intact. The patient, however, underwent reoperation because of valve dehiscence from atrial wall 6 months after surgery. Infectious abscess at the native mitral annulau was observed to be completely cured, and a mitral prosthesis was implanted at the annulus. The patient is discharged under satisfactory condition. PMID- 7474581 TI - [Thoracoscopic partial lung resection using staplers and polyglycolic acid (PGA) felts in an 85-year-old patient]. AB - An 85-year-old man was admitted to our hospital with complaints of dyspnea and palpitations. His chest roentgenogram and CT scan showed emphysematous lung and a right pneumothorax with bulla. In cases of emphysematous lung disease, it is well known that postoperative pulmonary leakage tend to be prolonged, and that control of such leakage is often difficult. A felt-like bioabsorbable prosthesis (polyglycolic acid sheet: NEOVEIL) was used for partial lung resection under thoracoscopic guidance. On each side of the stapler, PGA strips (1 cm wide x 3.5 cm long) were fixed with absorbable sutures. We found that it was difficult to cut with the stapler knife and that the second staple line could not easily cut across the first. However, there was no postoperative air-leakage and his postoperative course was uneventful. PMID- 7474582 TI - [Combination therapy with arbekacin and fosfomycin against postoperative severe mixed-pneumonia of MRSA in primary lung cancer patients]. AB - We experienced successful treatment of postoperative severe pneumonia of Methicillin-resistant Staphylococcus aureus (MRSA) with combination therapy of Arbekacin (ABK) and Fosfomycin (FOM) in three lung cancer patients. Case 1 was a advanced age of seventy-nine man who had had right upper lobectomy. Case 2 was a 61-year-old man who had had left lower lobectomy and extended bilateral mediastinal lymph-node dissection through the median sternotomy. And case 3 was a 59-year-old man who had suffered from pulmonary embolism after right pneumonectomy and partial resection of left atrium and superior vena cava. All cases were immuno-compromised patients and super-infected with Gram-negative rods, and Pseudomonas aeruginosa in case 1 and case 3. Clinical symptoms were improved after the start of administration of ABK and FOM inspite of ineffectiveness of prior treatment with other antibiotics. We added staggered chemotherapy of Sulbactam/Cefoperazone (SBT/CPZ) and Ceftazidime (CAZ) for case 1 and case 3 respectively. Thus, the combination therapy of ABK and FOM might be useful for severe pneumonia of MRSA in the immunocompromised patients, and the combined staggered chemotherapy of beta-lactum agents and above would be the first choice in the treatment for the case involving Pseudomonas aeruginosa. PMID- 7474583 TI - [Surgical management of vascular ring in 5 cases]. AB - Five cases of vascular ring are presented. Three cases have double aortic arch and two cases have right aortic arch with aberrant left subclavian artery. Other congenital heart diseases are complicated in four cases. MRI was useful for the evaluation of tracheal and esophageal compression. PMID- 7474584 TI - [Surgical treatment of incomplete endocardial cushion defect in adult patients]. AB - Seven adult patients (> or = 40 years old) and six pediatric patients with incomplete endocardial cushion defect (ECD) underwent corrective surgery in our institution. Preoperative catheterization study showed that the left-to-right shunting rate and the pulmonary-to-systemic flow ratio were greater in adult patients than those in pediatric patients, but systolic pulmonary arterial pressure was not high and similar in the two groups. The degree of mitral regurgitation by left ventriculography was mild or moderate in both groups. However, in adult patients, a variety of arrhythmias were observed. Postoperatively, NYHA grades and arrhythmia improved markedly in the adult patients. Surgical correction for ECD is recommended even in adult patients before aggravation of cardiac failure and arrhythmia. PMID- 7474585 TI - [A case report of prosthetic valve replacement for malfunction of the Hancock valve in mitral position associated with recurrent peptic ulcer and renal dysfunction]. AB - A 62-year-old woman was admitted with malfunction of the Hancock valve in mitral position. She had been suffering from gastroduodenal ulcer for about ten years. She couldn't take warfarin after 5 years later of the initial operation due to recurrent gastrointestinal bleeding. Judging from her age and renal dysfunction, we preferred mechanical valve to avoid the risks for the reoperation. After confirming the healed ulcer with administering omeprazole, we performed prosthetic valve replacement with SJM 29 M successfully. Postoperative course was uneventful and recurrence of the ulcer was not observed. PMID- 7474586 TI - [A case of Ebstein's anomaly treated by tricuspid valve replacement]. AB - A 8-year-old girl with Ebstein's anomaly was operated on because of progressive heart failure caused by severe tricuspid regurgitation. At the operation, the tricuspid leaflets were found to be destroyed, and the annulus was very enlarged. The enlarged true tricuspid annulus was constricted using the DeVega method, and the tricuspid valve was replaced on the true annulus using a porcine valve. To avoid injury to the auriculoventricular bundle, the suture line was shifted to the false annulus at the portion close to the course of His' bundle. Postoperative course was uneventful in satisfactory condition. The role of valve replacement in the treatment of Ebstein's anomaly of the tricuspid valve is not only to improve valve formation, but also to secure the function of the right ventricle. It is ideal to replace the tricuspid valve on the true annulus provided that the injury to the His' bundle is safely avoided. PMID- 7474588 TI - [Coronary artery bypass grafting for a patient with left ventricular ejection fraction of 16%: a case report and review of the Japanese literature]. AB - A 65-year-old man with left ventricular ejection fraction (LVEF) of 16% successfully underwent coronary artery bypass grafting (CABG). CABG used to be contraindicative for patients with LVEF below 20%. However, such patients are recently involved to indication of CABG if viability of the myocardium is proved. We consider CABG is indicated for a patient who keeps a sufficient cardiac output despite a low level of LVEF. Intraaortic balloon pumping which was started immediately after the induction of anesthesia was effective. Only eight patients with LVEF below 20% have undergone CABG in Japan. Thus, we reported our patient with some discussion on operative indication and perioperative management. PMID- 7474587 TI - [A case of type A acute aortic dissection successfully treated with a ringed intraluminal graft: a new technique of graft insertion]. AB - A-77-year old woman was hospitalized at our hospital presenting with severe chest and back pain. A computed tomographic scan revealed acute type A aortic dissection and intraoperative ultrasound showed an entry near the brachiocephalic artery. Selective cerebral perfusion using flexible 12 Fr balloons was performed for brain protection. The distal aorta was trimmed just proximal to the entry using felt strips and a 24 mm ringed intraluminal graft (ILG) was inserted under it successfully. The patient did well after the operation. Insertion of a ringed ILG is a simple and easy technique, however if an entry is located near the aortic arch, insertion is difficult because of intimal retraction to the arch. And an additional intimal tear sometimes occurs at the site of tape ligation. The method which we presented seemed to provide safer insertion of the proximal ring without these problems. PMID- 7474589 TI - [Two cases of left atrial myxoma originating from mitral annulus]. AB - We experienced two cases of left atrial myxoma originating from mitral annulus. Mitral valve replacement and mitral valvulo-annuloplasty were performed in each case, because of the presence of mitral leaflet defect due to removal of the myxoma. Two dimensional echocardiography was a useful method to confirm the location and decide the surgical approach. PMID- 7474590 TI - [Aortic valve replacement for rheumatoid aortic valve regurgitation]. AB - Valve replacement for aortic regurgitation in patient with rheumatoid arthritis was rare in Japan. We report a successful case in which aortic regurgitation necessitated aortic valve replacement. A 62-year-old woman was admitted for shortness of breath, chest pain and progressive edema. She had been treated for rheumatoid arthritis for more than ten years using steroids. The aortic valve was successfully replaced by a prosthetic mechanical valve. The histopathological examination of the excised aortic valve demonstrated rheumatic nodules in the right- and non-coronary cusp. It was supported that these changes caused shrinkage of cusp and resulted in aortic regurgitation. This patient was discharged on the 20th postoperative day. There was no evidence of detachment of the prosthetic valve. PMID- 7474591 TI - [A trial of antegrade perfusion to the lower half of the body with selective cerebral perfusion for repair of aortic arch aneurysm combined with atherosclerosis obliterance]. AB - A 73-year-old man with aortic arch aneurysm combined with atherosclerosis obliterance underwent aortic arch replacement. As he had bilateral femoral artery obstruction, we instituted ECC with arterial perfusion through the ascending aorta. Then, the left ventricle started dilating due to mild degree of aortic regurgitation. So with selective cerebral perfusion, we perfused the lower half of the body through a Foley balloon catheter which was inserted from the aortic arch antegradely. We could perfuse with the catheter for 20 minutes for core cooling and then during repair until the catheter disturbed the operation. The Foley catheter had given us some difficulties for handling during the procedure. Based on this experience, we have made a catheter which had both a balloon for aortic occlusion and a tube for perfusion and which we could keep away from operative fields, similar to the way we used antegradely insertable aortic occlusion balloon catheter. PMID- 7474592 TI - [A case of left atrial myxoma with remarkable elevation of serum alkaline phosphatase level]. AB - A 53-year-old woman was admitted suffering from high grade fever. Laboratory findings revealed leukocytosis, high CRP, high erythrocyte sedimentation rate and remarkable elevation of serum alkaline phosphatase level. Echocardiogram demonstrated left atrial tumor which was later diagnosed as myxoma. Although remarkable elevation of serum alkaline phosphatase level often indicates malignant disease, tumor makers were normal level and whole body computerized tomography couldn't detect other noncardiac diseases. Because of patient's deterioration due to sustained high grade fever, the left atrial tumor was excised. After the excision of the left atrial myxoma, patient's clinical signs and symptoms disappeared, and the serum alkaline phosphatase level was gradually normalized. We conclude that the cause of remarkable elevation of serum alkaline phosphatase level was the left atrial myxoma. PMID- 7474594 TI - [A case of primary cryptococcosis of the lung]. AB - A 37-year-old man was pointed out having an abnormal shadow in the right lung field at a medical checkup in July 1992. There was 5-year life history of rearing about 60 macaws since 10 years before. Upon operation a tumor 1.5 cm in diameter was palpated in the right S6. Intraoperative frozen section diagnosis revealed no malignancy. Partial excision was performed. The excised specimen showed a 1.5 x 1.4 x 1.0 cm, well-defined and yellowish white tumor. Histologically it was granuloma with a thin fibrous capsula, and most of the granuloma necrotized involving cryptococcus formans. Postoperative serum cryptococcus antigen and antibody titers were negative. There has been no sign of recurrence, as of 8 months after the operation. No adjuvant chemotherapy has been needed. PMID- 7474593 TI - [A technique for removal of entrapped intra-aortic balloon catheter through the left axillary artery]. AB - A case of intra-aortic balloon pumping (IABP) entrapment in which the ruptured catheter could be removed through the left axillary artery was reported. A 68 year-old man was admitted with persistent chest pain, and because of his poor cardiac condition, an IAB catheter was inserted by the Seldinger technique. During attempted extraction of the IAB catheter resistance was encountered and it turned out to be entrapped. Then a guide-wire was passed through the IAB lumen to the left axillary artery, the artery was exposed, and the tip of the guide-wire and IAB catheter were led to the arteriotomy site. After the balloon was dissected, the IAB catheter could be removed through the axillary artery. Finally, left femoro-left branchial artery bypass was performed with e-PTFE graft because of the arterial injury and ischemia of the involved arm. The patient recovered from this episode. PMID- 7474595 TI - [A case of localized fibrous mesothelioma arising from visceral pleura of the right lower lobe]. AB - A 56-year-old male who had been followed for chronic hepatitis had cough and hemosputum. Chest X-ray and CT films showed an abnormal mass shadow in the right thorax. On suspicion of intrathoracic tumor, resection was performed. The tumor, which was on the diaphragm without invasion, was pedunculated and arose from the bottom surface of visceral pleura of the right lower lobe. From the microscopic findings and immunohistochemical staining, the tumor was diagnosed as localized fibrous mesothelioma. Some localized fibrous mesothelioma exhibit as malignant tumor. Therefore, complete resection and intensive follow-up should be done. PMID- 7474596 TI - [Lung cancer manifested as cardiac tamponade: a case report]. AB - A case of lung cancer manifested as cardiac tamponade was reported. A 64-year-old male had anterior chest pain and dyspnea. A chest X-ray films showed an enlarged heart with a solitary round shadow in the left lung hilus. Echocardiogram disclosed a large amount of pericardial effusion. Emergency pericardiectomy with drainage was accomplished. The patient lived for 8 months without effusion accumulation. Histologically adenocarcinoma of the lung with prominent lymph nodes metastases was disclosed. Surgical decompression such as pericardiectomy or pericardial fenestration is a valuable selection in the treatment of pericardial effusion and tamponade. PMID- 7474597 TI - [A resected case with lung cancer rapidly progressed to the mediastinum]. AB - A 66-year-old man was revealed an abnormal shadow in the chest roentgenogram. Definitive diagnosis was not obtained by chest CT and bronchofiberscopy. Three months later, the shadow showed a rapid increase. We made a diagnosis of lung cancer by needle biopsy. Extended radical operation for the tumor was performed. The tumor with invasion to the chest wall and the mediastinum was completely resected with reconstruction of the chest wall and SVC. Postoperative pneumonia and respiratory failure were occurred, and patient died four months later. PMID- 7474599 TI - [Effect of acupuncture on the overactive bladder]. AB - BACKGROUND: We examined the effect of acupuncture for the overactive bladder. METHODS: Eleven patients (9 males and 2 females) with the overactive bladder were treated with acupuncture. The age of the patients ranged from 51 to 82 years (mean 71 years). Nine patients complained of urge incontinence and 2 patients of urgency. Uninhibited contraction was observed in all patients before the acupuncture. A disposable needle (0.3 mm in diameter, 60 mm in length) was inserted into bilateral BL-33 points at the depth of 50 to 60 mm and was rotated for 10 minutes manually. The treatment was performed 4 to 12 (average 7 times). RESULTS: Urge incontinence was controlled completely in 5 and partially in 2 of 9 patients. In 2 patients who complained urgency complete response was obtained after the treatment. Uninhibited contraction disappeared in 6 patients after the treatment. Acupuncture induced an increase of maximum bladder capacity and bladder compliance with statistical significance (p < 0.01 and p < 0.05), respectively. CONCLUSION: Acupuncture at the BL-33 point was effective for controlling the overactive bladder. PMID- 7474598 TI - [Surgical treatment of advanced renal cell carcinoma]. AB - BACKGROUND: We studied the indication and the limitation of surgical treatment for advanced renal cell carcinoma (RCC) based upon the survival rate for 48 patients (recurrent in 37, stage 4B in 11) who received the surgical therapy for the metastatic lesions. METHODS: We examined many factors including performance status (P.S.), laboratory findings, initial stages, surgical curability, operated organs, combined treatments with operation and grade of primary lesion in subjected patients. RESULTS: Regarding the performance status (P.S.) of the patients, those with P.S. 0 showed a favourable survival compared with those with P.S. 1 and 2, and significant differences were observed. Furthermore, the patients with P.S. 1 showed a favourable survival compared with those with P.S. 2, and a significant difference was observed between these two groups. The patients who manifested 2 or more abnormalities of the acute phase reactants showed unfavourable survival compared with those who manifested 1 or less abnormalities of the acute phase reactants, and a significant difference was observed between these two groups. As to the stage of the patients with stages 1 and 2 at their initial presentation, the patients who received the surgical treatment showed a favourable survival compared with those who did not, and significant differences were observed. As to the patients with stage 4B, although their clinical background was quite different, the patients who received the surgical treatment also showed a favourable survival compared with those who did not, and a significant difference was observed. In an analysis of the curability obtained by surgical procedures, the patients who were thought to have the curable surgery showed a favourable survival compared with those who were thought to have non-curable surgery, but no significant difference was observed between these two groups. In an analysis of the organs excised as a treatment for metastasis, the patients who underwent the operation for the affected adrenal gland and lung showed a relatively favourable survival. On the other hand, the patients who underwent the operation for the contralateral adrenal gland and pancreas showed an unfavourable survival. Regarding the combination treatment with operation, the patients who received the interferon and interleukin-2 treatment showed a favourable survival. No significant difference was observed among any combination treatments if operation was omitted from the combination. Regarding the grade of primary lesion, no consistent trend was observed. CONCLUSION: These results suggest that the surgical treatment should be indicated based upon the P.S. of the patients, the findings of acute phase reactants and the stage at the initial presentation of patients. Furthermore, the metastatic organs and the application of cytokine must be considered in the treatment of advanced RCC. PMID- 7474600 TI - [Clean intermittent catheterization in neurogenic bladder patients with vesicoureteral reflux]. AB - BACKGROUND: For neurogenic bladder (NGB) patients with vesicoureteral reflux (VUR), renal deterioration constitutes a primary threat to survival. Although anti-reflux surgery for those patients was reported to be effective, the efficacy of clean intermittent catheterization (CIC) in such patients still remains to be clarified. METHODS: Sixteen neurogenic bladder patients with VUR were treated with CIC. Eight of them had spina bifida and 3 had radical hysterectomy for uterine cancer. Other 5 patients had spinal cord injury, spondyrocace, measles encephalopathy, and 2 unknown diseases, respectively. Hyperactive bladder was noted in 11 patients, whereas hypoactive bladder was noted in 3. Three patients were not evaluated. RESULTS: VUR was improved in only 3 patients (3 ureters), whose VUR grades were not more than III. Nine patients (13 ureters) had to have antireflux surgery. Although 3 of them needed bladder augmentation afterwards, the success rate of the antireflux surgery was 84.6%. CONCLUSION: We concluded that CIC alone was not effective to control VUR in neurogenic bladder patients. Nevertheless, CIC remained a good treatment option when VUR was managed surgically. PMID- 7474601 TI - [The relationship between serum PSA value and the weight of the inner gland of the prostate in the cases of benign prostatic hyperplasia]. AB - BACKGROUND: To study reasons why serum PSA value (Markit-F) elevates in benign prostatic hyperplasia (BPH). METHODS: The relationship between total prostatic weight and the outer gland weight in 48 cases of BPH undergoing subcapsular prostatectomy was studied by ultrasound measurement. PSA was compared with prostatic weight, inner gland weight, outer gland weight, growth speed of the inner gland and age in the 48 cases. RESULTS: 1) The weight of the outer gland increased slightly in proportion to the development of BPH. However, the increase was not significant statistically. 2) PSA correlated with the inner gland weight and growing speed of the inner gland well. 3) PSA did not correlate with the outer gland weight and age. CONCLUSIONS: It is well known that PSA elevates not only in the cases of prostatic cancer but also in the cases of BPH and that PSA correlates with the whole prostatic weight. This study revealed that the elevation of PSA in the cases of BPH was caused by enlargement of the inner gland. PMID- 7474603 TI - [Clinical characteristics of 7 renal cell carcinoma patients developing a solitary pancreatic metastasis after nephrectomy]. AB - BACKGROUND: A solitary metastasis from renal cell carcinoma is rarely occurred evidence. Therefore, we intended to study on the clinical characteristics of those patients who developed a solitary pancreatic metastasis after nephrectomy. METHODS: Of seven hundred and fifty-two patients with renal cell carcinoma (RCC) who had been treated during the past 38 years, 7 patients developed a solitary pancreatic metastasis after nephrectomy. We studied on the clinical characteristics, treatment modalities and prognosis of these 7 patients. RESULTS: Regarding the age distribution at the time of nephrectomy, the median age was 48 years old. Therefore, the median age of patients developing a pancreatic metastasis was 10 years younger than that of the whole group of RCC. Regarding the sex ratio of these 7 patients, there was no difference between the male and female group (male in 4, female in 3). Therefore, relatively higher rate of pancreatic metastasis in female patients was observed compared with the male counterpart. Furthermore, no difference in the affected side of the primary tumour was observed in these 7 patients (left in 4, right in 3). Regarding the stages at the time of nephrectomy, a majority of patients (85.7%) showed low stages (stages 1 and 2). As to the grade of primary lesion, a majority of these tumours (85.7%) showed low grades (grades I and II). In an analysis of the interval from the time of nephrectomy to the pancreatic metastasis, the median interval was 140 months. Therefore, the percentage of the late recurrent patients was 71.4%. As to the site of metastasis in the pancreas, various sites in the pancreas was affected, with dominant site being its body. The median size of metastatic lesion was 4.25 cm. Regarding the treatment of pancreatic metastasis, resection of the metastasis was performed in 5 patients, and the remaining 2 patients received chemotherapy in combination with administration of interferon. As to the clinical courses of these 7 patients, other organs became involved with metastases in 6 patients, and 5 of them died with the median survival of 18 months. Furthermore, of the remaining 2 patients, one survived for 73 months with persistent cancer, and the other survived for 4 months without cancer. CONCLUSION: In conclusion, renal cell carcinoma accompanied by later development of pancreatic metastasis tend to occur in younger patients and have higher incidence in females than in males when compared with otherwise similar renal cell carcinomas. Furthermore, they tend to be of low stage and grade, have longer intervals to the occurrence of metastasis and carry a poor prognosis. PMID- 7474602 TI - [Clinicopathological evaluation of etoposide or estramustine phosphate in castrated patients with advanced prostatic cancer]. AB - BACKGROUND: We conducted a multicentric randomized trial to compare bilateral orchiectomy versus bilateral orchiectomy plus etoposide or estramustine phosphate as first-line therapy for advanced prostatic cancer (stage D2). METHODS: From January 1991 to December 1992 a total of 46 newly diagnosed cases (registered cases) of advanced (stage D2) prostatic cancer was randomized into 3 groups as follows; Group A: bilateral orchiectomy and 25 mg/day of etoposide every 2 weeks for 6 months. Group B: bilateral orchiectomy and 560 mg/day of estramustine phosphate for 6 months. Group C: bilateral orchiectomy alone. One of group A and one of group B were ineligible cases, so 44 were eligible. In the eligible cases, ages were ranged from 54 to 90 (mean of 71.2) years old. No significant difference of patients' characteristics was found among 3 groups and median follow-up period was 25 months. Response was evaluated based on the response criteria according to Japanese urological association. Specifically, a central pathologist who blinded to the treatment was employed for evaluating pathological response at six months. RESULTS: Of the 44 eligible patients, 33 and 25 were evaluated for clinically and pathological analyses, respectively. Clinical response rates were 80% (12/15) of group A, 100% (4/4) of group B and 78.6% (11/14) of group C. No significant difference in the clinical response and survival rate was shown among the three groups. Significantly higher frequencies of side effects were noted in the grop B compared to the other two groups (p < 0.05) and cardiovascular complications were the most frequent in group B. Favorable pathological response was obtained in all of group B, but not statistically significant compared with 7/21 (33.3%) of response rate in group A and C. The pathological response was significantly correlated with the clinical one in all patients (p < 0.01). While 8 of 11 patients (73%) with pathological response grade 1, 2 and 3 achieved clinical PR (partial response) or CR (complete response), only 5 of 14 (36%) with grade 0 received PR or CR. CONCLUSIONS: We conclude that low dose administration of etoposide or estramustine phosphate dose not improve clinical response and survival in a short term in castrated patients, but increases the adverse effects due to the drugs in these patients. In addition, the pathological evaluation at 6 months after treatment appears to reflect the clinical response at that time in newly diagnosed patients with advanced prostatic cancer. PMID- 7474604 TI - [Clinical significance of proliferating cell nuclear antigen (PCNA) and argyrophilic nucleolar organizer region (AgNOR) in testicular tumors]. AB - BACKGROUND: Proliferative potential of testicular tumor was assessed by immunohistochemistry using anti-proliferating cell nuclear antigen (PCNA) monoclonal antibody and silver staining of argyrophilic nucleolar organizer region (AgNOR), and the results were compared with clinical course. METHODS: Fourty-five patients with testicular tumor and 10 normal testicles were investigated. All specimens were fixed with 10% buffered formaliun within 24 hours. The interval between patient's recognition of testicular swelling for the first time and the date for operation was designated as M (months), and the specimen weight was designated as g (gram). Growth rate of the primary tumor was estimated roughly by g/M. RESULTS: PCNA positive rates in seminomas, non seminomas and normal testes were 70.5 +/- 19.1% (mean +/- S.D.), 80.4 +/- 10.5% and 17.7 +/- 7.8%, respectively. PCNA positive rate in stage I seminoma (64.4 +/- 19.9%) was significantly lower than stage II and III seminoma (83.6 +/- 7.3%) (p < 0.05), but not significant between stage I non-seminoma and stage II, III non seminoma. The mean numbers of AgNOR per nucleus in seminomas, non-seminomas and normal testes were 8.09 +/- 1.35 (mean +/- S.D.), 6.89 +/- 1.43, 4.18 +/- 1.60, respectively and significantly higher in testicular tumors than normal testes. There were, however, no significant difference between their clinical stages. Logarithmic significant correlation was observed between PCNA positive rate and the growth rate in primary lesion, but no correlation was found between mean number of AgNOR per nuclei and the growth rate. Of the 10 patients with stage I non-seminomas received surveillance policy, four relapsed. Of the 10 patients, 4 of the 6 patients with higher than 75% PCNA positive rate relapsed, whereas, none of the 4 patients with lower than 75% PCNA positive rate relapsed. There were no significant correlation between PCNA positive rate and number of AgNOR. CONCLUSION: PCNA is more accurate indicator of growth potential in testicular tumor than AgNOR and may predict recurrence in stage I non-seminomatous testicular tumor patients followed by surveillance policy. PMID- 7474605 TI - [Benefits and adverse effects of post-operative radiation therapy after radical cystectomy for patients with advanced bladder cancer]. AB - BACKGROUND: The benefits and adverse effects of post-operative irradiation for advanced bladder cancer patients were investigated. METHODS: Ten patients with pT3b, pT4 or pN+ bladder cancer who underwent radical cystectomy at Yamanashi Medical University hospital during 7 years and 3 months from October 1983 to December 1991 received adjuvant chemotherapy and radiotherapy (Group 1). During the same period, six patients with recurrent tumor after radical cystectomy were treated by radiotherapy (Group II). Stages of the primary tumors were pT2 in 1, pT3a in 2, pT3b in 6 and pT4 in 7 cases. In addition, 10 of 16 patients (63%) had positive nodes. RESULTS: During the follow up period, seven patients died of cancer, and one died of other cause. As a result eight patients (5 in Group I, 3 in Group II) are alive. The cumulative 5-year survival rate is 50%. However, nine of the 16 patients (56%) suffered from the small bowel obstruction as an adverse effect of irradiation. Six patients required resection of the small bowel or bypass surgery. CONCLUSIONS: Radiation after radical cystectomy seemed to be effective for the local control of the tumor, but the adverse effect to the digestive system was very severe and common. PMID- 7474606 TI - [5-Fluorouracil and alpha-2a interferon in patients with hormone-refractory prostate cancer]. AB - BACKGROUND: Hormone-refractory metastatic prostate cancer remains a disease for were limited therapeutic options are available. Therefore, the establishment of newly, more effective chemotherapy is expected. Experimental data suggest that PC 3, a human hormone refractory prostate cancer cell line, showed a 2-fold increase in 5-Fluorouracil (5FU) sensitivity in the presence of alpha-2a Interferon (IFN alpha 2a) at 100 IU/ml, compared to that without IFN alpha 2a. Based on this data, we treated 11 patients with 5FU and IFN alpha 2a in order to determine the clinical response and toxicity of this combination chemotherapy. METHODS: One course of this combination chemotherapy consisted of a continuous intravenous infusion of 5FU at 600 mg/m2/day for 5 days (D1-D5) with IFN alpha 2a 3 million units (MU) intramuscularly 3 times weekly (D1, D3, D5) followed by a bolus injection of 5FU at 600 mg/m2 and IFN alpha 2a at 3 MU/day on D15 and D22. RESULTS: Based on the Response Criteria for Prostate cancer Treatment, one of 9 patients with bony metastasis had partial response, 2 patients with nodal disease on the CT scan obtained partial response. Six of 11 patients had more than 50% decrease in post-therapy prostatic antigen levels, 3 of whom obtained complete response. Significant myelosuppression did not occur. There were no chemotherapy related deaths. CONCLUSION: These results suggest that the combination of 5FU and IFN alpha 2a, although preliminary, is an active regimen against hormone refractory metastatic prostate cancer. PMID- 7474607 TI - [New nocturnal penile tumescence recorder]. AB - BACKGROUND: Records of nocturnal penile tumescence are still important test and necessary to distinguish organic impotence from psycogenic impotence. METHODS: We developed the nocturnal penile tumescence recorder with a disposab indiumgallium straingauge, and we examined its clinical usefulness for volunteers with normal erectile function. RESULTS: The results were that the indium-gallium straingauge had almost the same resistance change with extension, and that it showed almost the same resistance value at respective circumferences with consecutively ten times measurements. Only two of thirty-nine volunteers experienced the snapping of the indium-gallium strain-gauge. None of the volunteers complained of discomfort during measurements. The nocturnal penile tumescence of five volunteers were recorded with a conventional mercuric straingauge and a newly developed indium-gallium straingauge simultaneously, and there were no significant differences between the results of the two materials. This recorder is very simple to manage, and the softwares for preservation of data and making graph are also simple. This new straingauge is clean and dose not require maintenance because of the disposable use. CONCLUSION: We concluded that this recorder system was clinically useful for t examination of nocturnal penile tumescence. PMID- 7474609 TI - [A study on the relationship between sensitivity to CDDP and inducibility of metallothionein and glutathione of genitourinary tumors]. AB - BACKGROUND: In this paper we demonstrate the relationship between the antitumor activity of cis-diamminedichloroplatinum (II) (CDDP) and inducibility of metallothionein (MT) and glutathione (GSH) of genitourinary tumors. METHODS: The chemosensitivity test was performed in athymic mice bearing tumors derived from the human tumor cell lines: ACHN (renal cell carcinoma), NMB-1 (urinary bladder transitional cell carcinoma), and NMT-1 (testicular embryonal cell carcinoma). A single dose of CDDP (25 mumol/kg body weight), was administered i.p. to athymic tumor bearing mice. Concentrations of platinum, MT, and GSH were measured in organ and tumor homogenates 24 h after CDDP administration. RESULTS: We observed that tumors derived from NMB-1 and NMT-1 were very sensitive to CDDP, but ACHN derived tumors were resistant to CDDP. Measurement of platinum concentrations in tumor tissues revealed no correlation to the observed chemosensitivities of the tumors. Furthermore, 24 h after CDDP administration, the levels of MT and GSH in NMB-1 and NMT-1 derived tumors were lower them or equal to those of control mice. In contrast, mice bearing tumors derived from ACHN exhibited a 1.7-fold and a 2.1 fold increase in MT and GSH, respectively, as compared to control mice. CONCLUSION: These findings suggest that the inducibility of MT and GSH in tumor tissues following CDDP administration may be a contributing factor in the development of CDDP resistance in renal cell carcinoma. PMID- 7474610 TI - [A case of sarcomatoid carcinoma of the urinary bladder]. AB - We reported a case of sarcomatoid carcinoma of the bladder in a 62-year-old man admitted to our hospital because of intermittent macroscopic hematuria. Under the diagnosis of invasive bladder tumor, the patient underwent radical cystectomy and construction of an ileal conduit. Microscopic examination showed a tumor composed of spindle cells that are arranged in a storiform pattern reminiscent of that seen in malignant fibrous histiocytoma without any component of obvious epithelial tumors. Immunohistochemical examination showed that the spindle cells were positive for cytokeratin and we diagnosed the tumor as sarcomatoid carcinoma. The patient has remained well without any evidence of recurrence for 30 months after radical cystectomy. The distinction between sarcomatoid carcinoma that lacks conventional epithelial component and MFH is very difficult and important. PMID- 7474608 TI - [The adrenoceptor and calcitionin-gene related peptide receptor in the striated urethral sphincter in male rabbit]. AB - BACKGROUND: The objective of this study is to analyze the adrenoceptor and calcitonin-gene related peptide receptor in the isolated striated urethral sphincter from a male rabbit. METHODS: The striated urethral sphincter preparations were suspended in 2ml tissue chamber filled with Krebs Ringer solution, and the changes of isometric twitch contraction induced by electrical field stimulation (EFS-contraction) were recorded in the presence of 3-isobutyl-1 methyl-xanthine (IBMX, 10(-5) M). RESULTS: The EFS-contraction was almost completely attenuated by tetrodotoxin (TTX, 10(7) M), vecronium (10(-4) M) and suxamethonium (10(-4) M). Norepinephrine (NE, 10(-8) M-10(-4) M) did not affect the EFS-contraction, but increased the tonic contraction in a dose-dependent manner. The tonic contractions induced by NE was significantly blocked by phentolamine (10(-6) M). Clonidine (10(-7) M), yohimbin (10(-7) M) and propranolol (10(-9) M-10(-6) M) did not affect the EFS-contraction. Isoproterenol (10(-9) M-10(-6) M) did not reduce the carbachol (10(-5) M) induced tonic contraction. Calcitonin-gene related paptide (CGRP, 10(-7) M-3 x 10(-6) M) did not affect the EFS-contraction and did not increase the tonic contraction. CONCLUSION: These results suggested that alpha 1-adrenoceptors of the striated urethral sphincter play a role to modulate the resting tension level, but alpha 2 adrenoceptors, beta-adrenoceptors and CGRP receptors play no role in this regard. PMID- 7474611 TI - [A case of urachal carcinoma associated with elevated levels of CA19-9]. AB - A 53-year-old man was admitted with the chief complaint of gross hematuria. Cystoscopic examination revealed a non-papillary tumor at the dome of the bladder. The plasma level of CA19-9 was markedly elevated before treatment. Total cystectomy with en bloc removal of the tumor and radical lymphadenectomy were performed under the diagnosis of urachal carcinoma. The plasma CA19-9 level decreased to the normal level after operation and was well correlated with postoperative course. The monitoring of CA19-9 may be useful for follow up in patients showing a high elevation of CA19-9. PMID- 7474612 TI - [A retroperitoneal cyst with elevated level of CA19-9 in its content]. AB - A 52-year-old man, who had been treated for left mega-hydronephrosis (5.6L) 25 years ago in our hospital, recently complained of left lumbago and fever. An abdominal CT scan revealed a cystic mass partially surrounded by calcification in the retroperitoneal space, which appeared to be a hydronephrosis. Percutaneous puncture of the cystic mass was performed and 670 ml, brownish-yellow purulent fluid containing a high level of CA19-9 (over 10,000 U/ml) was obtained. Cytologic diagnosis of the fluid revealed class II cells. Within 7 months after the puncture, recurrence developed twice. Percutaneous instillation of 95% ethanol or Minocycline hydrochloride was instilled, but neither of them prevented recurrence. Ultimately, excision of the cystic mass was performed. Histopathological findings of the cystic mass showed no evidence of malignancy. The patient is follow-up with periodic CT scan showed no evidence of recurrence for more than 3 years. PMID- 7474613 TI - [Influence of pneumoperitoneum on respiratory function during urological laparoscopic surgery]. AB - PURPOSE: To determine the influence of pneumoperitoneum on respiratory function during urological laparoscopic surgery. MATERIALS AND METHODS: We performed laparoscopic surgery on 82 patients, 45 of these patients were examined in detail of the effect of pneumoperitoneum on the respiratory function. Of these 45 patients, 14 patients received lymphadenectomy and 31 patients received adrenalectomy. For comparative analysis, 4 patients with laparoscopic cholecystectomies were also examined. Under general anesthesia, carbon dioxide tension (PaCO2) was analyzed before, during, and after the pneumoperitoneum. The data were analysed along with 4 factors; the patient's spirographic parameters, alveolar-arterial gas difference (AaDO2), insufflation pressure for the pneumoperitoneum, and operating time. RESULT: After starting the pneumoperitoneum, PaCO2 levels rose significantly. The PaCO2 levels of patients underwent laparoscopic adrenalectomy were higher than that of patients underwent laparoscopic cholecystectomy. The preoperative vital capacity and forced expiratory volume in one second (FEV1.0) did not show a significant difference of increase in the degree of PaCO2 increase. In contrast, the important factors that were closely related to the elevation of PaCO2 were preoperative AaDO2 and intraoperative insufflation pressure. Patients with high preoperative AaDO2 (> 10 torr) showed a significant increase in PaCO2 levels after pneumoperitoneum. When insufflation pressure was high (> = 10 mmHg), there was a significant rise of PaCO2 after pneumoperitoneum. In all patients whose operating time was 300 minutes or more, high levels of PaCO2 continued on the next day after surgery. CONCLUSION: We should still carefully observe the insufflation pressure, operating time, and other ventilation conditions during laparoscopic surgery. PMID- 7474614 TI - [A study of calcium metabolism in calcium-containing stone formers]. AB - To clarify the pathogenesis of calcium-containing urinary stone, calcium metabolism of the 113 patients with calcium-containing stone was studied by fasting and oral calcium loading test. The stone patients were classified into four groups. Those were normocalciuria (NC, n = 60), absorptive hypercalciuria-1 (AH-1, n = 26), absorptive hypercalciuria-2 (AH-2, n = 16) and renal hypercalciuria (RH, n = 11). In the AH-1 group, hypercalciuria resulted from enhancement of intestinal calcium (Ca) absorption. The increase in serum Ca from absorbed Ca increased renal filtered load of Ca. Urinary excretion of Ca was correlated to that of sodium (Na) in fasting and Ca load, and both increments were also correlated in Ca load. As of this fact, the increase in Na excretion was responsible for a cause of hypercalciuria in the AH-1 group. In the RH group, the serum ionized Ca level and % tubular reabsorption of Ca were significantly lower than those of other groups. Urinary excretion of Ca and Na ratio was significantly higher than that of the NC and AH-1 groups. This condition was due to an impaired renal tubular reabsorption of Ca and the resulting secondary hyperparathyroidism. These findings suggested was response disorder of distal tubular to parathyroid hormone. Urinary excretion of phosphorus (P) was correlated to that of Na in fasting and Ca load, and both increments were also correlated in Ca load in the AH-1 group. The increase in urinary excretion of P resulted from the secondary hyperparathyroidism in the RH group.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7474615 TI - [Prognosis after radical surgery in prostatic cancer patients with lymph nodes metastases]. AB - We investigated prognosis of clinically localized prostatic adenocarcinoma patients who revealed to have had lymph nodes metastases by undergoing radical surgery. Eighty six patients were operated during the last 15 years under the clinical diagnosis of A2, 9 patients, B1, 15 B2, 13 and C, 49, respectively. Total prostatectomy was done to 51, total cystoprostatectomy to 33 and total pelvic excentration to 2 patients. Of these patients, 22.2% with stage A2, 20.2% with B1, 7.7% with B2 and 43.8% with C had positive nodes and the rate of positive nodes in stage C was significantly higher than that in other stages (p < 0.01). Regarding histological differentiation, 15.4% of well, 23.7% of moderate and 51.6% of poor by differentiated had positive nodes and the rate of positive nodes in poor by differentiated was significantly higher (p < 0.01). In 2 of 21 cases whose lymph nodes were dissected to the level of the aortic bifurcation, positive nodes were detected only in the external and common iliac areas. These two cases were missed, i.e., "false negative" if limited nodes dissection was performed. All patients with positive nodes were treated with hormonal therapy. The 5-year cancer specific survival rate of patients with positive (n = 27) and negative (n = 59) nodes were 66.4% and 92.4%, respectively. The prognosis of patients with positive nodes were significantly worse than that of patients with negative nodes (p < 0.001). Among 27 patients with positive nodes, significant prognostic factor was not number or extent of positive nodes, but histological differentiation.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7474616 TI - [Clinical study on stage PT1 and grade 3 transitional cell carcinoma of the urinary bladder]. AB - Thirty-three patients who had transitional cell carcinoma of the bladder with stage PT1 and grade 3 components were treated between January 1980 and December 1992. Clinical study was done for these 33 patients on treatment modalities and it's problem, intravesical recurrence, disease progression and prognosis. Twenty five of 33 patients underwent bladder-sparing surgeries and total cystectomy was performed for another 8 patients as initial treatment. Intravesical recurrence was observed in 18 (72%) of 25 patients who underwent initial bladder-sparing surgeries. Recurrence-free rate was 44% for 1 year and mean period to recurrence from initial therapy was 13.1 months. Although some adjuvant therapy after initial treatment was performed for 20 patients, it's efficacy in preventing intravesical recurrence could not be confirmed. Disease progression was recognized in 10 (40%) of 25 patients initially treated by bladder-sparing surgeries after median latent period of 33.7 months, whereas only one (12.5%) of another 8 patients who underwent total cystectomy showed progression, and 5-year progression-free rate after initial bladder-sparing surgeries was 55.6%. Three and five-year actural survival rate in all cases were 81.5% and 65.1%, respectively. According to initial treatment, 5-year survival rate was 62.2% for the patients treated by bladder-sparing surgeries, whereas it was 83.3% for the cases who underwent total cystectomy. However statistical significant difference was not recognized. In view of disease progression after initial bladder-sparing surgeries, patients with progression yealed a 45.7% 5-year survival rate compared to 81.5% for the patients without progression, and significant difference was demonstrated between them.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7474617 TI - [A study of middle-high aged impotent patients]. AB - Background factors, which are causes of functional and organic impotence, of 729 impotent patients over 50 years old were evaluated. They were classified into 3 major categories, i.e., stresses at job, stresses at home, and diseases or accidents. Over 30% of the patients had 2 or more categories. A retirement from office and troubles at job were most frequent among stresses at job. As to stresses at home, marital problems such as wife's death and remarriage were most common. With respect to diseases or accidents, they were observed in 84 percent of all patients, and hypertension (HT) or diabetes mellitus (DM) were most common and the rate of medication was considerably high. The percent of clearly organic impotence was quite low (22%). Hormonal environment of 303 over 50 aged impotent patients was checked and compared with 120 impotent patients from 20 to 49 years old. Serum testosterone (T) levels in patients over 70 years of age decreased significantly. Lutenizing hormone (LH) and follicle stimulating hormone (FSH) levels in patients after age 50 progressively increased. Patients administered anti-androgenic agenst tended to show lower T and higher LH, FSH, and prolactin (PRL) levels than non-administered. Patients with psychotropic drugs showed significantly higher PRL levels. Hormonal therapy (mainly T replacement therapy) tended to be more effective in patients of low serum T levels before therapy. However, some patients with normal T improved. In 141 impotent patients, 83 cases of which were after age 50, the degrees of their penile arterial impairment were tested using penile brachial index (PBI) and pulse volume recordings (PVR). PVR waveforms were classified into 3 groups, i.e., normal, slightly abnormal, and markedly abnormal. PBI was significantly lower in abnormal groups than in normal group. Between each parameter of PVR and PBI, statistically significant correlation and relevancy were found. Crest time were significantly longer, and PBI and angle of rise significantly lower in over 50 aged patients than in 20-49 aged. We evaluated the risk factors to penile arterial impairment, such as DM, HT, smoking, and cardiovascular disorders. Each of these risk factors was minor to age factor itself. PVR proved to be useful, simple, and non-invasive method for the screening of vascular impotence. In conclusion, degenerative changes occur about hormonal environment and penile blood flow according to aging, and many kinds of background factors have direct or indirect influence to occurrence of impotence. We emphasize such matters should be considered at examination of middle-high aged impotent patients. PMID- 7474618 TI - [Treatment of advanced renal cell carcinoma with a combination of interferon alpha and gamma]. AB - A total of 29 patients with advanced renal cell carcinoma entered a pilot study of combination therapy with interferon alpha (IFN-alpha) and interferon gamma (IFN-gamma). IFN-alpha (HLBI: 3 x 10(6) IU, BALL 1:5 x 10(6) IU, IFN-alpha-2a: 9 x 10(6) IU or IFN-alpha-2b: 6 x 10(6) IU) was given intramuscularly every day and IFN-gamma (IFN-gamma-1a: 3 x 10(6) JRU) was given intravenously by drip infusion 3 times a week (every 2-3 days). The treatment was continued for 3 months as the induction therapy, and then the tumor response was evaluated. Of the 22 evaluable patients, 4 achieved a partial response (PR), 10 showed no change (NC), and in 8 the disease had progressed (PD) during the therapy. Thus, the overall response rate was 18.2% [95% confidence interval (CI) 2.1-34.3%]. A favorable response tended to be obtained in patients with good performance status or small pulmonary metastases, or in those who had no prior therapy with IFN-alpha, who received this treatment immediately subsequent to radical nephrectomy, or who received IFN gamma as much as possible according to this regimen. Toxicity was evaluated in 28 patients: fever, general fatigue, anorexia, leukocytopenia and impaired liver function were frequently noted, and 3 patients were withdrawn from the study because of such adverse effects. In patients who had a PR or NC, the same dosage of IFN-alpha was continued to be given intramuscularly 2-3 times a week (every 2 4 days) as the maintenance therapy.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7474619 TI - [Flow cytometric analysis of the DNA content of the urinary bladder cancers- study on the DNA heterogeneity in bladder cancers]. AB - PURPOSE: To evaluate the clinical relevance between the DNA ploidy and histopathology, and the incidence of the DNA heterogeneity in patients with bladder cancers. METHODS: Flow cytometry (FCM) was used to study the DNA ploidy in 63 patients who underwent total cystectomy. The DNA ploidy and DNA index were analyzed by FCM in total 328 paraffin embedded samples (5.2 samples per case on the average). RESULTS: The DNA ploidy of 52 bladder cancers, that had coexisted after total cystectomy, showed that 24 cases, 46% were DNA aneuploid and 18 cases, 35% had DNA heterogeneity. The DNA ploidy of 11 cases that were eradicated after cystectomy was all DNA diploid. There were significantly good correlation among DNA ploidy pattern and intravesical involvement (lymph duct involvement and venous involvement), but were not among the DNA ploidy pattern and tumor grade and stage. With regard to the evaluation of two vertical divided samples of tumors, DNA aneuploid had been not always recognized in the deeper sample, therefore, we did not determine that there was good correlation between the DNA ploidy and the tumor invasion. CONCLUSION: These data suggest that although the incidence of DNA heterogeneity in bladder cancers (35%) is thought to be relatively small, the DNA ploidy will be able to the important prognosticating factor in bladder cancers. PMID- 7474620 TI - [How does thermotherapy effectively work on benign prostatic hyperplasia--an experimental study]. AB - Isometric contractile force of rabbit prostatic tissue in response to electric field stimulation (EFS), KCl, and phenylephrine were measured at incubation temperature of 37 degrees C, before and after thermal exposure to 42 degrees C, 45 degrees C, 48 degrees C and 50 degrees C for 30 minutes. The contractile force in response to EFS decreased after thermal exposure above 45 degrees C, and the contractile force in response to KCL or phenylephrine decreased after thermal exposure above 48 degrees C. All the contractile response abolished after thermal exposure to 50 degrees C. The results indicate that the nerve is more hear sensitive than the smooth muscle in the prostate. Histological examination revealed shrinkage of cell body and dark staining of nuclear chromatin of the smooth muscle cells after thermal exposure above 48 degrees C. The same histological change of the smooth muscle as well as degenerative change of the nerve cells was observed on the prostate 3-7 months after clinical thermotherapy. From these results, it is suggested that clinical effect of thermotherapy is brought about from both neural and muscular damage of the prostate. Since the least temperature to cause an irreversible tissue damage ranges from 48 degrees C through 50 degrees C, we believe it is ideal to heat the prostate around 50 degrees C to obtain a good clinical effect of thermotherapy on benign prostatic hyperplasia as a minimum invasive treatment. PMID- 7474621 TI - [Value of PSA/gamma-Sm ratio (P/S ratio) for diagnosis of prostate cancer in patients with urinary retention]. AB - It is known that serum prostate specific antigen (PSA) in urinary retention due to benign prostatic hypertrophy (BPH) increase. To evaluate prognostic value of the ratio of serum PSA to gamma-seminoprotein (P/S Ratio) for prostate cancer (PC) in patients with urinary retention, we have studied the P/S Ratio at the initial examination in 33 patients with untreated PC (10 with and 23 without urinary retention) and 193 patients with untreated BPH (38 with and 155 without urinary retention) histopathologically diagnosed at our hospital between January, 1992 and December, 1993. The results were as follows: 1) The mean P/S ratio of PC patients was significantly higher than that of BPH patients in both groups with and without urinary retention. 2) When the cut off value of P/S Ratio was determined to be 1.35, the highest efficiency, 59.3% was obtained in the group without urinary retention. The sensitivity and specificity were 65.2% and 91.0%. 3) In the group with urinary retention, the efficiency was also the highest, 80.0% with a cut off value of 1.35. The sensitivity and specificity were 80.0% and 100%. 4) In all patients, the efficiency was 64.6%, the sensitivity was 69.7%, and the specificity was 92.7% with a cut off value of 1.35. 5) Positive rate of serum PSA in BPH patients with urinary retention was 47.4% and that in BPH patients without urinary retention was 17.4%. The mean P/S ratio of the BPH patients with urinary retention was significantly lower than that of BPH patients without urinary retention, which suggested that the serum free PSA increase in the former.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7474622 TI - [Clinical investigation of renal pelvic and ureteral cancer with special reference to adjuvant chemotherapy]. AB - Clinical investigation of 93 patients with histologically confirmed renal pelvic and ureteral cancer were performed. These patients consisted of 55 males and 38 females with a mean age of 64.8 years. There were 61 cases of renal pelvic cancer, 55 cases of ureteral cancer and 23 with cancers of both sites. Thirty four cases were associated with bladder cancer and 41 of 82 patients had multiple tumors. The overall 5-year survival rate was 46.0%. 5-year survival of stages pTa, pT1, pT2, pT3, and pT4, was 93.3%, 71.8%, 37.5%, 30.4% and 10.5%, respectively. In this report, we evaluated various prognostic factors according to the survival rate. Sex, age, tumor localization, multiplicity, associated bladder cancer and concomitance of CIS had no influence on survival. In the ABC analysis, the B group showed a tendency for a poor prognosis. However it may be explained from the fact that the B group contained more patients at advanced stages than the other groups. Tumor grade, tumor stage, pV factor and pL factor had a significant effect on survivals. But tumor grade, pV and pL factors were closely related to the tumor stage. Thus the stage was thought to be the most important factor in the prognosis of upper urinary tract cancer. Different surgical procedures and irradiation also did not affect the prognosis of the patients with the same degree of invasion. Chemotherapy for all stages had no effect on survivals compared with non-chemotherapeutic group. However only for pT3 and higher stage cases, cisplatin-based chemotherapy improved the prognosis compared with patients not given chemotherapy. In conclusion, chemotherapy containing cisplatin should be considered for treatment of high stage upper urinary tract cancer. PMID- 7474623 TI - [A clinical study of associated bladder tumor in patients with renal pelvic and ureteral tumor]. AB - We investigated the incidence of associated bladder tumor and prognosis in 101 cases with a pathological diagnosis of transitional cell carcinoma, selected from those with renal pelvic and ureteral tumor whom we had encountered over the 18 years between April 1976 and March 1993. Among these 101 cases, the incidence of associated bladder tumor was noted in 42 (41.6%), 23 (22.8%) with coexistence and 19 (18.8%) with subsequence. As for the primary site of renal pelvis and ureter, the coexistence was 15.4% and subsequence 20.5% in renal pelvis, and the coexistence was 24.6% and subsequence 19.3% in ureter, and the coexistence was 60.0% and subsequence 0.0% in both renal pelvis and ureter. The incidence of coexistent bladder tumor was high in both renal pelvis and ureter, but no significant difference was noted. As for the stage, the incidence of coexistence was high in T1, while subsequence was high in T2, but no significant difference was noted. As for the grade, the incidence of coexistence was high in G2, but no significant difference was noted. The 5 year survival rate was 58.2% in those without, 54.2% with coexistence, and 82.5% with subsequent bladder tumor, with a significant difference (p < 0.05) between the last two groups. The interval of subsequent bladder tumor ranged from 4 to 164 months (mean 27.7 months), with the incidence within 2 years being approximately 70.0%. It was found that the renal pelvic and ureteral tumors are frequently associated bladder tumor while associated bladder tumor dose not appear to have an ill effect on the prognosis. Therefore it is necessary that patients with renal pelvic and ureteral tumor be observed closely for 5 years, especially for the initial 2 years. PMID- 7474624 TI - [Urinary tract abnormalities associated with anorectal malformations]. AB - Anorectal malformation (ARM) is often associated with urological problems such as congenital urogenital anomalies, recto-urinary fistula, neurogenic bladder due to vertebral anomalies and operative complications. We analyzed 57 cases of ARM and discussed about the management of associated urogenital anomalies during neonatal and infantile period. The incidence of urogenital anomalies was 85.7% in high type, 65.5% in intermediate type and 38.1% in low type. Among these urinary tract anomalies, VUR was most common and was documented in 38.6% of ARM patients. Renal dysplasia, PUJ stenosis, megaureter and urethral stenosis was also common in these patients. Renal dysfunction was documented in 5 cases (2 in high type, 2 in intermediate and 1 in low type), mainly due to VUR and renal aplasia. These results show the need for evaluation of urinary tract during the neonatal and early infantile period even in low type ARM. The management of urinary tract anomalies associated with ARM is firmly related with the management of ARM itself, and we must be closely in co-operation with pediatric surgeons. PMID- 7474625 TI - [A case of huge renal hamartoma]. AB - We experienced a case of renal hamartoma of mesothelial origin. A 21-year-old was referred to our hospital with a complaint of growing abdominal mass. The diagnosis of right renal tumor was confirmed based on the radiographic examinations. The renal tumor was removed by partial nephrectomy. Histological examination revealed the tumor composed of fibroblasts, smooth muscle and a few tubule-like epithelial tissue. Thus, this case was classified as renal hamartoma of mesothelial origin. PMID- 7474626 TI - [Primary adenocarcinoma of the tunica vaginalis testis expressing CA19-9 antigen]. AB - A 47-year-old man presented with a 10-year history of left scrotal swelling. No solid mass detected on sonogram, dark-red colored fluid aspirated, equivocal cytology, and elevated serum CA19-9 level prompted surgical exploration. Frozen section reported no malignancy of thickening of the tunica vaginalis. The final pathology adenocarcinoma, however, necessitated left radical orchiectomy and hemiscrotal resection, demonstrating no malignancy at the left testis nor epididymis. CA19-9 level was high (104,200 U/ml) in the fluid obtained at the first surgery and the tumor cells were positively stained for CA19-9. Imaging study including abdominal CT scan and upper G-I series and normalization of CA19 9 level denied other tumor existing. In conclusion, this is to our knowledge the first case report of adenocarcinoma of the tunica vaginalis expressing CA19-9 antigen. PMID- 7474627 TI - [Squamous cell carcinoma of the ureter as a late complication of ureterocutaneostomy--a case report]. AB - A case of squamous cell carcinoma of the ureter was reported. A 62-year-old woman, who underwent left nephrectomy and right ureterocutaneostomy at the age of thirty-one because of tuberculosis of the urinary tract, visits our hospital periodically for an ureteral catheter change. An urgent nephrostomy was performed because of ureteral stricture on March 8, 1994. Three months later tenderness and induration appeared at the former ureteral stoma. The resected specimen was histologically diagnosed as squamous cell carcinoma. Right palliative ureterectomy did not produce the desired effects due to the invasion of the tumor into the surrounding organs. In this rare case, it is suggested that chronic infection and irritation of the indwelling catheter might be causative of squamous cell carcinoma of the ureter. PMID- 7474628 TI - [Predicting renal stone fragility against ESWL]. AB - BACKGROUND: The mineral content (MC) and mineral density (MD) of renal stones, measured by dual energy X-ray absorptiometry, were compared with the experimental and clinical results of stone disintegration by shock wave. METHODS AND RESULTS: In the experimental study, the MC and MD values correlated significantly with the total shock wave energy used, with a correlation coefficient (r) for MC being 0.814 and for MD, 0.706 (n = 46). The mean MC and MD values were 0.521 +/- 0.387g and 0.317 +/- 0.121 g/cm2 (n = 14) respectively in patients undergoing clinically successful ESWL, while those were 4.27 +/- 6.19g and 0.626 +/- 0.302 g/cm2 (n = 6) respectively in patients undergoing clinically unsuccessful ESWL. Those values differed significantly between the two groups (p < 0.05, p < 0.01, respectively). The combination of MC, MD and area values of a stone was shown to be the best predictor of treatment outcome, with a predictive value of incorrect discrimination being 17% and that of correct discrimination, 85% using linear discriminant analysis. CONCLUSION: The mineral content and mineral density, measured by dual energy X-ray absorptiometry, may provide the best prediction of outcome of patients undergoing ESWL treatment. PMID- 7474629 TI - [Clinical assessment for prognostic factors related to recurrence and progression in superficial bladder cancer]. AB - PURPOSE: The objective of this assessment is to identify prognostic factors of recurrence and disease progression of primary superficial transitional cell carcinoma of the bladder. PATIENTS AND METHODS: TUR-Bt was performed in 150 patients with initial superficial bladder cancer, of which pathological diagnosis was pTa or pT1 transitional cell carcinoma. The recurrence and progression free survival was examined. The clinicopathological factors analyzed were as follows: grade, pT, tumor size, tumor number and tumor form. RESULTS: Median follow-up period was 39 months (range 3-184). The age distribution was from 25 to 98 years old with the average of 67. The patients were 122 males and 28 females. Recurrence was observed in 72 patients. The 5-year recurrence free rate for all cases were 41.6%. The factors found to be of significance for the prognosis for recurrence were tumor size (1 cm <) and tumor multiplicity (p < 0.01). Progression was seen in 15 patients; 8 invasive tumors, 2 lymph node metastases and 5 distant metastases. Twelve cases of those died of cancer. The 5-year progression free rate for all cases were 87.0%. The prognostic factors related to progression found in this study were G3, pT1 and sessile form (p < 0.01). There was no significant difference in recurrence free rate between cases with and without postoperative therapy; UFT p.o., ADM i.i. (intravesical instillation), ADM + Ara-C i.i. and PEP + 5-FU + Ara-C i.i. CONCLUSION: Tumor size more than 1 cm and tumor multiplicity were the high risk factors about recurrence. So were G3, pT1 and sessile form about progression. PMID- 7474631 TI - [A study on cathepsin B-like substance in cancer of the urinary tract]. AB - Cathepsin B, a cysteine proteinase, has been shown to be increased in tumor tissue and to be responsible for the process of invasion and metastasis. To study the biochemical characteristics of cathepsin B-like substance in the tissue of a non-cancerous kidney and some urologic cancer tissues, we extracted cathepsin B like substances from a non-cancerous human kidney tissue, human renal tumor, human renal pelvic tumor and human bladder tumor. The cathepsin B-like substance extracted from the non-neoplastic human kidney has a molecular weight of approximately 28,000. On the result of western blotting with anti-human liver cathepsin B antibody, two types of protein were observed in the fraction extracted from urologic cancer tissues. The molecular weights of them were 28,000 and 35,000, respectively. Both types of cathepsin B-like substances had the same activities for benzyloxy-carbonyl-L-arginine-L-arginine-4-methyl-7-coumarylamide, and both activities were inhibited by (L-3-transcarboxyoxiran-2-carbonyl)-L leucyl-agmatin. These findings suggest that both cancer cells of the renal parenchyma and urothelium secrete cathepsin B-like protease which express not only a normal cathepsin B but also a higher molecular weight of cathepsin B-like protease and that a higher one plays an important role in a process of invasion. PMID- 7474630 TI - [Expression of GP160 mRNA in renal cell carcinoma: an attempt to predict the sensitivity of IFN-alpha therapy]. AB - BACKGROUND: Alpha-interferon (IFN-alpha) has significant antitumor activity against advanced renal cell carcinoma (RC). Although numerous clinical trials reported major responses in approximately 15% of patients subjected to INF-alpha therapy, it has been impossible to predict which patients will respond to IFN alpha. RC cell lines could be subdivided into those that are inhibited by IFN alpha and those that are not. The expression of GP160 (Aminopeptidase a/kidney associated differentiation antigens) correlated with resistance to IFN-alpha. In RC xenografts using a mouse model, though GP160(+) RC cell lines were resistant to IFN-alpha, GP160(-) RC cell lines were markedly sensitive to IFN-alpha. METHOD: In this study, we assayed 26 RC patients for the expression of GP160 in mRNA level using RT-PCR (reverse transcription polymerase chain reaction). RESULTS & CONCLUSION: In 23 (88.5%) patients GP160 mRNA expression are remarkable as in normal kidney tissue, but in 2 (7.7%) patients, GP160 mRNA expression was weak. And in 1 (3.8%) patient, GP160 mRNA expression was hardly detectable, and in this patient complete remission was achieved with IFN-alpha therapy for advanced lung and mediastinal metastatic RC. PMID- 7474632 TI - [Flow cytometric analysis of DNA content of urinary bladder cancers--study on primary tumors and metastasized lymph nodes]. AB - PURPOSE: This study was designed to evaluate DNA ploidy patterns and metastatic patterns between primary tumors and metastatic lymph nodes in bladder tumor patients with lymph node metastases. METHODS: Flow cytometry (FCM) was used to study the DNA ploidy. The DNA ploidy patterns in 16 lymph node metastases in relation to the degree of ploidy in the primary bladder tumor were evaluated in 63 patients who underwent total cystectomy. RESULTS: The primary tumor that had metastasized was G3 tumor in grade and over pT2 in stage in many cases. Thirty nine diploid tumors had given raise to lymph node metastases in only 5 cases (13%), whereas 11 cases (46%) of aneuploid tumors had metastasized (p < 0.01). With regard to ploidy patterns between primary tumors and the corresponding lymph node metastases, four patterns were noted, namely D-->D (5 cases), D + A-->D (4), A-->A (5) and A-->D (2) (D: DNA diploid, A: DNA aneuploid). The DNA index between the primary tumors and the corresponding lymph node metastases was the same in all but 2 cases (14/16.88%). In cases with lymph node metastases, the prognosis was very poor whether or not the DNA ploidy of the primary tumors or the metastatic tumors was DNA aneuploid. CONCLUSION: These data suggest that a malignant cell on the primary tumor metastasized to the lymph node in many cases. PMID- 7474633 TI - [A semi-quantitative measurement of glycocalyx and an ATP bioluminescent assay for the analysis of Pseudomonas aeruginosa biofilm]. AB - BACKGROUND: In order to analyze each element of a bacterial biofilm, a semi quantitative measurement of glycocalyx using toluidine blue and an improved ATP bioluminescence assay were developed. RESULTS: In the toluidine blue assay, a linear correlation was obtained between optical density and the amount of gellan gum, which is an exopolysaccharide derived from Pseudomonas elodea. By using the improved ATP assay, ATP at concentration of 1 x 10(-4) mol/l or floating bacterial cells of 10(3) cfu could be detected. The applicability of these two methods was examined in evaluating a total amount of glycocalyx and ATP content of Pseudomonas aeruginosa biofilms, which were developed or treated with antimicrobial agents in a modified Robbins device. CONCLUSION: The changes in optical density and ATP luminescence were reasonable, and well correlated with those obtained by scanning electron microscopy. These results indicate that the toluidine blue assay and ATP assay are useful in analyzing bacterial biofilms. PMID- 7474634 TI - [Blood levels of cyclosporine, acute rejections and the prognosis of the allografts in pediatric renal allograft recipients]. AB - BACKGROUND AND METHODS: Results of a total of 46 pediatric (mean age; 9.0 years), living-related, cyclosporine-treated renal allograft recipients were analyzed retrospectively to study 1), the effects of an acute rejection on the long-term allograft function and 2), the relationship between the cyclosporine blood levels and the frequency of acute rejections. In addition, sequential allograft biopsies (100 days, one, two and three years post-transplant) were performed in all cases to see if clinical symptoms correlated with histological changes. RESULTS: After a mean follow up of 3.3 (range 1-6) years, 15 of 22 (68%) recipients who had no signs of acute rejection, 6 of 12 (50%) who had acute rejections but responded completely to therapy, and none of the remaining 12 cases who failed to recover completely from acute rejections showed good allograft functions and normal histology on biopsy. Thirty-four of 46 cases had no clinical signs of rejection at 100-day allograft biopsy. Yet, histological changes compatible with acute rejection were noted in 11 of 34. Treatments of the rejection were promptly started on these patients and the prognosis was excellent in six cases. The mean blood trough levels of cyclosporine were significantly lower in patients who developed an acute rejection during the first week following transplantation as compared to those without rejection episodes during the same period, 165.5 ng/ml and 204.1 ng/ml, respectively. Furthermore, the mean blood cyclosporine trough levels in patients who experienced rejections during two, three and five weeks after transplantation were definitely lower than those who did not experience such episodes; 124.5 ng/ml, 101 ng/ml and 149.3 ng/ml versus 241.7 ng/ml, 217.7 ng/ml and 201.1 ng/ml respectively, although the number of the cases was too small to reach statistical significance. CONCLUSION: The results indicate that the long-term allograft function would depend on whether there were episodes of acute rejections, and/or how the allograft responded to the treatments. Since the introduction of cyclosporine has made it difficult to detect acute rejections by clinical findings alone such as blood chemistry or urinalysis, the role of routine allograft biopsies is very important in renal transplantation. It seems likely that low blood levels and/or acute decline of cyclosporine during early postransplant period could increase the possibility of episodes of acute rejection. Since it is well documented that cyclosporine induces less renal parenchymal damages in children, the blood through levels of the drug should be kept at between 200 and 300 ng/ml within 5 weeks of transplantation in order to reduce the risk of acute rejections and to improve the chance of long-term allograft survival. PMID- 7474635 TI - [Comparisons of histological grading between biopsy and radical prostatectomy specimens in prostatic adenocarcinoma]. AB - PURPOSE: To examine how accurately biopsy specimens reflect the corresponding excised specimens in terms of histologic grades of prostatic adenocarcinoma, histologic grades of biopsy specimens were compared to those of radical prostatectomy specimen in 29 patients who underwent radical prostatectomy without any prior treatment. METHODS: For the assessment of histological grading, Japanese General Rule for Prostatic Cancer (JGRPC), in which adenocarcinoma was graded as being either well, moderately or poorly differentiated, and Gleason's system were applied. RESULTS: According to JGRPC, histologic grades of biopsy specimens were identical to those of the prostatectomy specimens in 83% of cases, with biopsy specimens given higher grades than those of the prostatectomy specimen in 4 (14%), and lower grades in 1 (3%). According to Gleason score histological grade of biopsy specimens were identical to those of the prostatectomy specimens in only 52%. However, the grouping category of Gleason score (2 to 5, 6, 7 and 8 to 10) assigned to the biopsy was lower than that of prostatectomy specimen in only 3 cases (10%) and was greater than that of prostatectomy specimen in 5 cases (17%). CONCLUSION: These results suggest that histological grades based upon biopsy specimens will predict those of the corresponding prostatectomy specimens in the majority (72% to 83%) but not all of the cases. PMID- 7474636 TI - [A study on international prostate symptom score in the assessment of therapeutic effects and severity of symptoms due to benign prostatic hyperplasia]. AB - BACKGROUND: Benign prostatic hyperplasia (BPH) is a common disease among elderly men in Japan. Recently, various non-invasive treatment modalities have been developed, however the criteria for the diagnosis and therapeutic effects of BPH have not been established yet. World Health Organization sponsored International Prostate Symptom Score (I-PSS) was devised. The criteria, however, were limited to evaluate the frequency of the subjective symptoms of BPH. In addition, this criteria did not include the quality of life (QOL) score. PATIENTS AND METHODS: We examined I-PSS and a new by modified symptom score for patients who underwent TUR-P. RESULTS: The efficacy of TUR-P could be confirmed by I-PSS, however the criteria proposed in I-PSS and in our modified symptom score were shown to be inadequate to evaluate QOL, and to be not specific scale for BPH. It was demonstrated that objective findings such as peak flow rate, residual urine volume and prostate volume did not correlate with the QOL score, however each objective and subjective significantly improved after TUR-P. CONCLUSION: These results suggested that further effort should be made to establish reliable criteria for the diagnosis and evaluation of therapeutic efficacy considering QOL in BPH patients. PMID- 7474637 TI - [Expression of proliferating cell nuclear antigen (PCNA) in renal cell carcinoma]. AB - PURPOSE: The objective of this study is to evaluate the relationship among PCNA positive ratio, pathological findings, nuclear DNA contents, and prognosis in renal cell carcinoma. Immunohistochemical analysis using the monoclonal antibody of PCNA were performed on a total number of 151 formalin-fixed paraffin-embedded samples (1-7 samples with a mean of 3.8) from 40 renal cell carcinomas. The percentage of PCNA positive cancer cells to the total amount of cancer cells was expressed as labeling index (LI; %). By means of flow cytometry we analysed nuclear DNA contents from the same samples. RESULTS: 1) LIes in pathological grades were 2.79 +/- 3.33% (mean +/- SD) for grade 1 (n = 16), 5.63 +/- 4.08% for grade 2 (n = 20), and 9.95 +/- 4.59% for grade 3 (n = 4). There was significant difference between grade 1 and grade 3 (p < 0.05). 2) LIes in pathological stage were 3.22 +/- 3.05% for pT2 (n = 22) and 7.01 +/- 4.99% for pT3 and pT4 (n = 18) (p < 0.05). 3) There was no significant correlation between LI and lymph node involvement. And there was no significant correlation between LI and distant metastasis, either. 4) LIes in nuclear DNA contents were 2.41 +/- 3.14% for DNA diploid (n = 17) and 6.80 +/- 4.29% for DNA aneuploid (n = 23) (p < 0.05). 5) It was suggested that LI was shown to vary according to the parts in a given tumor examined in parallel with tumor heterogeneity of nuclear DNA content. 6) The 5 year cause-specific survival rate of the patients with LI > or = 5.0% (n = 15) was 40%, while that with LI < 5.0% was 74% (n = 25) (p < 0.05). CONCLUSION: These findings suggest that LI, which was determined by immunohistochemical analysis using PCNA antibody indicates a growth potential of renal cell carcinoma and is available for estimating malignant potential. PMID- 7474638 TI - [An analysis of familial prostate cancer]. AB - BACKGROUND: Clinical and pathological features of familial prostate cancer patients were examined. METHODS: We compared 7 pairs (14 patients) of familial prostate cancer patients (Group F) with those of 1741 prostate cancer patients diagnosed in around Gunma Prefecture between 1987 and 1993 (Group G). Relative survival rates were compared because the average age was different in two groups. RESULTS: In Group F and Group G, age at diagnosis ranged from 54 to 86 and 47 to 97 years old, and the averages were 68.1 +/- 8.5 (S.D.) and 74.2 +/- 8.3, respectively. The average age tended to be younger in Group F. The ratio of early stage cancer tended to be higher and that of poorly differentiated cancer tended to be lower in Group F. The 3- and 5-year relative survival rate was 82.4% and 57.6% in Group F, and 84.3% and 73.9% in Group G, respectively. The 5-year survival rate tended to be lower in Group F, but almost no difference was observed between two groups on the whole. In Group F, of 6 patients whose causes of death were clear, 4 (66.7%) died of prostate cancer, while in Group G, of 398 patients whose causes of death were elaar, 224 (56.3%) died of prostate cancer. Family history of other cancer sites was positive in 3 of 6 families in Group F. CONCLUSIONS: Familial prostate cancer patients tended to be young, in early stage and low ratio of poorly differentiated cancer. PMID- 7474639 TI - [Continuous subcutaneous injection therapy with interferon-alpha for renal cell carcinoma patients with bone metastasis]. AB - Thirteen patients with renal cell carcinoma who had proven bony metastases were treated with multimodal treatment including surgery, radiotherapy and immunotherapy in the form of subcutaneous continuous injection of by natural type interferon-alpha (INF). The mode of administration of IFN was as follows: IFN, 2,5000 x 10(4) unit dissolved in 60 ml saline, was continuously injected (0.5 ml/hr) via a subcutaneous route as one course of the treatment and was given two courses in two weeks preoperatively. Postoperatively, IFN was given every week and the number of courses totally amounted to 15. In some cases IFN was given thereafter either every week or every other week. In four patients whose serum concentration of IFN was measured during and after administration of continuous IFN, the concentration of IFN rose after injection and showed 40.5 IU/ml in average 24 hours later. The concentration was kept measurable in six to eight days long and the maximum concentration was 167 IU/ml. In IFN-treated patients nine survived including two CRs, two NCs, five PDs and four deaths. The five year survival rate was 53%. Continuous subcutaneous injection of IFN in combination with surgery and/or radiotherapy is effective in the treatment of bony metastasis from renal cell carcinoma. PMID- 7474640 TI - [Tuberculous spondylitis after intravesical BCG instillation: a case report]. AB - We report a case of tuberculous spondylitis after intravesical bacillus Calmette Guerin (BCG) instillation. A 71-year-old man was administered BCG (80 mg per week) for 8 weeks for prophylactic treatment of bladder cancer. After the first instillation he experienced miction pain, pollakisuria, and febrile episodes. Two months after the completion of BCG instillation, he complained to back pain and spinal X-ray showed a lytic lesion of Th7 vertebra. Diagnosis of metastatic transitional cell carcinoma was made based on MRI and bone scan. But pathological findings at laminectomy revealed tuberculous spondylitis. Antituberculous therapy (SM, RFP, and INH) was instituted and anterior supine fusion was performed. Now he is free from bladder cancer and tuberculous infection. Intravesical BCG instillation is effective for superficial bladder cancer, but it should be kept in mind that complications related to this treatment could occur and the adequate antituberculosis treatment has to be insisted if indicated. PMID- 7474641 TI - [Primary seminoma of the prostate and embryonal cell carcinoma of the left testis in one patient: a case report]. AB - A 27-year-old Japanese male presented with gross hematuria and right frank pain, and was diagnosed of having a right ureteral stone by IVP. Though the stone was passed spontaneously, gross hematuria continued, and his urinary stream became week. The prostate was moderately enlarged. The prostate biopsy was initially thought to represent prostate rabdomyosarcoma. He underwent a pelvic extenteration followed by chemotherapy with dacarbazing and ADM. Three years after the pelvic exenteration, coin lesions were found on a chest X-ray film. He was diagnosed as embryonal cell carcinoma of the left testis with pulmonary metastasis and received 3 courses of PVB therapy. At this time, the former diagnosis of prostatic rabdomyosarcoma was changed to prostatic rabdomyosarcoma was changed to prostatic seminoma, because placental-ALP staining was positive. This patient had two distinct germinal cell tumors in the prostate and the left testis. He was alive without any tumor recurrence 20 months after PVB chemotherapy. To the best of our knowledge our report represents the first patient of seminoma of the prostate in the world. PMID- 7474642 TI - [Radical prostatectomy in a patient with hemophilia B]. AB - We describe our experience with radical prostatectomy in a patient with hemophilia B. whose factor IX activity was 7% and partial thromboplastin time was 55.9 sec. A total of 43,000 units of factor IX concentrate (PCC) was given perioperatively for 2 weeks. No abnormal hemorrhage was noted. An algorithm for perioperative management of hemophilia B patient is discussed. PMID- 7474643 TI - Influence of nocodazole on the development of donor blastomeres from 16-cell stage bovine embryos in nuclear transfer. AB - The aim of the present study was to establish a reliable procedure with nocodazole treatment for the synchronous cleavage of blastomeres of bovine embryos used as nuclear donors for nuclear transfer. Sixteen-cell stage embryos derived from in vitro-maturation, fertilization and culture were used. In three initial experiments, embryos were incubated in mTCM-199+FCS with various concentrations (0-20 microM) of nocodazole under 5% CO2 in air. The concentrations required to arrest the blastomeres in the mitotic phase were examined. The effects of 10 microM nocodazole were also examined by observation of the division rate of blastomeres after the removal of nocodazole. Ninety percent (90%) of the blastomeres were arrested in the mitotic phase when embryos were exposed to 10 and 20 microM nocodazole. Exposure to 10 microM nocodazole had the highest blastomere-cleavage rate (47%). When the exposure period to 10 microM nocodazole was prolonged to 36 hr, the division rate of the blastomeres decreased. Furthermore, the effects of 2 culture conditions (mTCM-199 under 5% CO2 in air vs modified synthetic oviduct fluid medium under 5% CO2, 5% CO2 and 90% N2) were compared on the division rate of blastomeres of embryos exposed to 10 microM nocodazole for 12 hr. When the embryos were exposed to nocodazole in mSOF, the division rate of blastomeres was improved to about 60%. The blastomeres produced by this treatment condition were used as nuclear donors and the developmental potential of the reconstituted embryos was investigated. The developmental rate to the blastocyst stage was 30.1% (58/193). Five embryos were transferred to 5 recipient cows and 2 of the 5 recipients (40%) became pregnant. Subsequently, one normal calf was born. PMID- 7474644 TI - Preliminary efficacy trial of Cymelarsan in mice artificially infected with Trypanosoma brucei brucei isolated from a dog in Zambia. AB - An efficacy trial of Cymelarsan on a Zambian strain of Trypanosoma brucei brucei was done. Twenty-five male mice were infected intraperitoneally with 10(6) of T. b. brucei isolated from a dog. Five groups of 5 mice were treated with 0 (control), 0.25, 0.5, 1.0 and 2.0 mg/kg Cymelarsan, respectively. The target was to achieve aparasitaemia for 30 days post-treatment, euthanising those that remained parasitaemic or relapsed before then. The 0.25 and 0.5 mg/kg groups remained parasitaemic although the parasitaemic levels were reduced. The 1.0 mg/kg group had a proportion of aparasitaemic mice. However, all mice in the 2.0 mg/kg group remained aparasitaemic until day 20 when 2 mice relapsed. These results suggested that more than 2.0 mg/kg was required to eliminate this strain. PMID- 7474645 TI - Suspected nutritional myopathy in two captive Bennett's wallabies (Macropus rufogriseus). AB - Between January and April, 1992, two captive Bennett's wallabies (Macropus rufogriseus) from a wildlife park in Iwamizawa City, Hokkaido, Japan, were diagnosed as having nutritional myopathy. One wallaby, four-year-old adult male, had mild fresh lesions, confined to the skeletal muscles, with poor cell reaction. The other wallaby, 8-month-old female joey, had marked and extensive lesions with cellular reactions and regenerative muscle fibers in the locomotor muscles. Involvement of other striated muscles such as those of the tongue, diaphragm and heart was mild with minimal cellular reaction. Age susceptibility may account for the difference in the extent of severity of the lesions between these two animals. PMID- 7474646 TI - How to differentiate ejection murmurs from systolic regurgitant murmurs. AB - Although it is not possible to distinguish the shape of a murmur as being crescendo-decrescendo as in ejection murmurs from a plateau-shaped regurgitant systolic murmur by hearing the murmur alone, it is possible to distinguish them by auscultation with the use of other characteristics that give clues to their shape or their origin. With the use of accompanying heart sounds, the peak of the crescendo-decrescendo effect of ejection murmurs makes a recognizable rhythm which is absent in the plateau-shaped regurgitant murmur. When no heart sounds are present, the increase in loudness after a sudden long diastole of only the ejection murmur can help to separate the two types of murmur. The term "midsystolic" as applied to the ejection murmur is misleading and should be avoided because it is based on controversial hemodynamics and it obstructs teaching the significance of early, mid, and late peaks to ejection murmurs. PMID- 7474647 TI - Collagen metabolism in the fibroblasts derived from the skin of a 16-year-old patient with Marfan syndrome: a decreased rate of intracellular degradation of collagen. AB - The rate of collagen synthesis in the cell layer fraction of the skin fibroblasts derived from a patient with Marfan syndrome was greater than that in the normal fibroblasts. Neither the relative content of type III collagen nor the level of prolyl and lysyl hydroxylations in type I collagen changed. Intracellular degradation rate of collagen in Marfan syndrome fibroblasts was decreased to one fifth of that in the normal fibroblasts. These results suggest that abnormal collagen metabolism may be related to the pathogenesis of clinical manifestations of Marfan syndrome. PMID- 7474648 TI - Management of acute pancreatitis: critical assessment. PMID- 7474649 TI - Growth hormone and calcitriol as modifiers of bone formation in renal osteodystrophy. PMID- 7474650 TI - Apoptosis, nucleosomes, and nephritis in systemic lupus erythematosus. PMID- 7474651 TI - NF-kappa B and transcriptional control of renal epithelial-inducible nitric oxide synthase. AB - Expression of the inducible isoform of nitric oxide synthase (iNOS) is subject to strict tissue specific transcriptional control. Recently, the NF-kappa B/Rel family of transcription factors, and particularly c-rel, was shown to mediate bacterial lipopolysaccharide (LPS) induction of iNOS in macrophages. Since LPS is only a weak inducer of iNOS in most nonimmune cells, we investigated the role of NF-kappa B in the regulation of iNOS expression in mouse renal epithelial cells. We report that LPS activates NF-kappa B in renal epithelium, but that this is not sufficient for induction of iNOS activity. The NF-kappa B complexes activated by LPS in renal epithelium differ from those in macrophages in that they lack c-rel, which may explain the absence of iNOS induction in renal epithelium. Conversely, LPS and interferon-gamma (IFN) synergize to induce renal epithelial iNOS. Functional iNOS promoter analysis indicate that this synergistic induction requires NF-kappa B. We conclude that NF-kappa B is necessary but not sufficient for the induction of renal epithelial iNOS expression, and that in contrast to macrophages, c-rel does not appear to play a major role in the regulation of renal epithelial iNOS. PMID- 7474652 TI - High glucose augments angiotensin II action by inhibiting NO synthesis in in vitro microperfused rabbit afferent arterioles. AB - Preglomerular afferent arteriole (Af-Art) is a crucial vascular segment in the control of glomerular hemodynamics. We have recently reported that vascular reactivity of Af-Art is modulated by nitric oxide (NO). However, little is known about its reactivity under pathophysiological conditions such as diabetes, which is often accompanied by abnormal glomerular hemodynamics. In the present study, we examined the direct effects of high glucose, the hallmark of diabetes, on the vascular reactivity of Af-Art. Rabbit Af-Arts were microperfused for three hours with medium 199 containing either normal (5.5 mM; NG-Af-Arts) or high concentrations (30 mM; HG30-Af-Arts) of glucose, and then vascular reactivity was examined. Sensitivity to angiotensin II (Ang II) was significantly higher in HG30 Af-Arts than in NG-Af-Arts. Ang II began to cause significant constriction from 10(-9) M in NG-Af-Arts (18 +/- 3%, N = 6, P < 0.01) and from 10(-11) M in HG30-Af Arts (9 +/- 2%, N = 6, P < 0.01). NO synthesis inhibition with 10(-4) M nitro-L arginine methyl ester (L-NAME) increased the sensitivity to Ang II in NG-Af-Arts without affecting Ang II action in HG30-Af-Arts. In L-NAME-pretreated NG-Af-Arts, Ang II began to cause constriction from 10(-11) M (11 +/- 3%, N = 6, P < 0.01). Thus, pretreatment with L-NAME abolished the difference in sensitivity to Ang II between NG- and HG30-Af-Arts, suggesting impaired NO synthesis in HG30-Af Arts.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7474653 TI - Cellular localization of tissue kallikrein and kallistatin mRNAs in human kidney. AB - The renal kallikrein-kinin system has been implicated in the regulation of blood pressure and sodium/water excretion. The activity of renal kallikrein is controlled by a number of factors in vivo. Kallistatin is a newly identified serine proteinase inhibitor (serpin) which binds to tissue kallikrein and inhibits its enzymatic activity in vitro. To understand the role of kallistatin in modulating tissue kallikrein's function in vivo, we examined the anatomical relationship between human tissue kallikrein and kallistatin in the kidney by in situ hybridization histochemistry. Tissue kallikrein and kallistatin gene transcripts were identified using digoxigenin-labeled riboprobes at the cellular level. Antisense and sense riboprobes corresponding to the 3' region of the human kallikrein and kallistatin mRNAs were synthesized by in vitro transcription and used for hybridization. Using an antisense kallikrein riboprobe, sites of kallikrein synthesis were localized in the distal tubules, collecting ducts and Henle's loops of the kidney. To a lesser degree, juxtaglomerular cells were also stained. Kallistatin mRNA was found at the same sites where kallikrein mRNA was localized. The most intense signals of both kallikrein and kallistatin were seen in the distal tubules and collecting ducts. Hybridization was specific for the target mRNA since sense kallikrein or kallistatin riboprobe did not bind to the sections. Immunoreactive human renal kallikrein and kallistatin levels were measured in the kidney and urine by immunoassays using specific antibodies. Co localization of kallikrein and kallistatin mRNA in the kidney suggests a potential role of kallistatin in regulating tissue kallikrein's function. PMID- 7474654 TI - Tumor necrosis factor-alpha augments the pro-inflammatory interaction between PMN and GBM via a CD18 dependent mechanism. AB - Acute glomerulonephritis is frequently associated with intraglomerular neutrophil (PMN) accumulation and the intensity of the inflammatory reaction is correlated with elevated concentrations of pro-inflammatory cytokines such as tumor necrosis factor-alpha (TNF alpha). PMN are thought to damage glomeruli due to a combination of reactive oxygen species and proteolytic enzymes. Using an in vitro model of anti-GBM nephritis the effects of TNF alpha on GBM damage by PMN were evaluated. The interaction of GBM and PMN resulted in a low grade respiratory burst that was significantly augmented by the addition of TNF alpha. Luminol dependent chemiluminescence (LCL) was increased from 2.4 x 10(6) to 48.1 x 10(6) (P < 0.05). The GBM induced LCL could be > 85% inhibited by blocking with monoclonal antibodies (mAbs) to the common beta chain of the PMN beta 2 integrin family (CD18), but was unaffected by mAbs to CD11a or CD11b subunits. Degradation of GBM, however, was not influenced by either TNF alpha priming of PMN or anti beta 2 integrin mAbs. When PMN were incubated with GBM-anti-GBM IgG complex they underwent an increase in LCL from 2.4 x 10(6) to 31.1 x 10(6). They also degraded more GBM than controls (10.1% vs. 1.8%). These aspects of PMN activation were Fc receptor mediated, dependent upon anti-GBM IgG being bound to GBM and inhibited by mAb to the PMN Fc receptor. These studies show that TNF alpha can modulate the inflammatory response of PMN in contact with GBM in a CD18 dependent manner. In contrast, Fc receptor mediated events are uninfluenced by TNF alpha. PMID- 7474655 TI - Human IgG anti-DNA antibodies deposit in kidneys and induce proteinuria in SCID mice. AB - We investigated the capacity of five human monoclonal IgG anti-DNA antibodies derived from lupus patients to produce glomerular immune deposits. The hybridomas secreting these antibodies were administered intraperitoneally to severe combined immunodeficiency (SCID) mice. Three of the five antibodies (B3, 35.21, 33.C9) were detected in the kidneys, but only one (33.C9) deposited exclusively in the glomeruli in the mesangium and capillary wall, whereas the other two antibodies bound to nuclei both in the kidney and in other organs. The antibodies were tested against a variety of autoantigens by ELISA, the only unique feature of 33.C9 was that it also bound strongly to histones. There were no particular amino acid motif that was related to immunoglobulin deposition in the kidney. All the mice that had immunoglobulin deposited in the kidney, either extracellularly or intranuclearly developed 2 to 3+ proteinuria, whereas the other mice had only trace amounts of proteinuria. This study demonstrates that some human monoclonal IgG anti-dsDNA antibodies are capable of binding to the glomerulus while others can penetrate cells and bind to nuclei in vivo. Although no abnormal pathology was observed, proteinuria was detected, perhaps representing an early phase of disease. These results indicate that the affinity for dsDNA is not the sole determining factor governing the biological properties of human anti-DNA antibodies in vivo. PMID- 7474656 TI - Effects of chronic hyperfiltration on proximal tubule bicarbonate transport and cell electrolytes. AB - The compensatory response to unilateral nephrectomy (UNX) was investigated by a combination of renal clearance, microperfusion, electron microprobe, and morphological techniques. Filtration rate was significantly elevated 21 days following UNX and associated with a marked stimulation of bicarbonate and fluid absorption in the proximal tubule. Analysis of kinetic data of bicarbonate transport demonstrated strong flow-dependent activation of bicarbonate absorption in both control and experimental condition. The bicarbonate level at which half saturation (apparent Kd) of transport occurred decreased uniformly at higher flow rates, but maximal transport rates (apparent Vmax) in the proximal tubule doubled in the remnant kidney. The flow dependence of bicarbonate transport in control and experimental conditions can be explained by an apparent unstirred layer effect modifying radial bicarbonate gradients in the tubule. Both Na/H-exchange and electrogenic H secretion contribute to bicarbonate absorption, but only Na/H exchange increased significantly in proximal tubules of UNX rats. Cell ion concentrations after UNX were unchanged in cortical tubules, consistent with proportionately enhanced apical and basolateral ion transport. Proximal tubule cell rubidium concentration measured after a 30-second rubidium infusion as an index of basolateral Na,K-ATPase activity was unchanged in UNX rats. Inasmuch as cell volume increased significantly (25%), these data are consistent with a proportionate and similar stimulation of rubidium uptake and Na,K-ATPase activity. PMID- 7474657 TI - Cultured proximal cells derived from transgenic mouse provide a model to study drug toxicity. AB - The effects of gentamicin on N-acetyl-beta-D-glucosaminidase (NAG) and acid phosphatase (AcP), two lysosomal enzymes present in proximal renal tubule cells, were studied in the PKSV-PCT cell line derived from proximal convoluted tubules from the kidney of a transgenic mouse carrying SV40 large T antigen under the control of the L-type pyruvate kinase gene. Gentamicin (400 micrograms/ml for 72 hr) did not alter cell viability, but significantly reduced cell growth and favored the formation of myeloid bodies. Gentamicin (50 to 800 micrograms/ml for 72 hr) decreased in a dose-dependent manner the cellular NAG in PKSV-PCT cells and stimulated its secretion by 20 to 60%. Chloroquine (50 to 100 microns) and ammonium chloride (NH4Cl, 30mM), two lysosomotropic amines known to stimulate the secretion of lysosomal enzymes in fibroblasts and macrophages, also stimulated secreted NAG in PKSV-PCT cells. However, the effect of chloroquine was less marked in PKSV-PCT cells than in cultured mouse 3T3 fibroblasts. Gentamicin induced lysosomal alkalinization but, in contrast to chloroquine and NH4Cl, the aminoside strongly stimulated the secretion of AcP. The secretion induced by gentamicin was nonpolarized, since the percentage of secreted NAG significantly increased from both the apical and basal sides of PKSV-PCT cells grown on permeable filters. Thus, these data suggest that gentamicin alters the secretion of NAG and AcP by a non-specific pathway and indicate that the PKSV-PCT cell line is a suitable system to examine the cellular action of drugs in kidney proximal tubule cells. PMID- 7474658 TI - An evaluation of the role of polyamines in different models of kidney hypertrophy in mice. AB - The role of ornithine decarboxylase (ODC) and polyamines in kidney hypertrophy is controversial. Since part of this controversy could be related to differences in the model system used by the different authors, we studied the changes in renal ODC and polyamines in six different models of kidney hypertrophy in mice, including compensatory renal hypertrophy produced by unilateral nephrectomy, experimental diabetes, potassium depletion and treatment with hormones such as testosterone, thyroxine and fluorocortisone. Only in the case of renal hypertrophy produced by testosterone administration was there a significant increase in ODC activity and putrescine content in the kidneys. However, the concomitant treatment with difluoromethylornithine (DFMO), an irreversible inhibitor of ODC, as a 2% solution in the drinking water completely abolished the increase of renal ODC, but the kidney weights increased and other androgenic effects, such as the induction of renal beta-glucuronidase, were not affected. Moreover, DFMO-treatment did not prevent the kidney enlargement produced in other types of hypertrophy, even in the cases associated with hyperplasia. The present results support the premise that, at least in mice, the increase in ODC activity and polyamine biosynthesis is not required for kidney growth, and also that in most cases renal enlargement is not accompanied by any increase in the polyamine content. PMID- 7474659 TI - TGF-beta and phorbol esters inhibit mitogenesis utilizing parallel protein kinase C-dependent pathways. AB - Transforming growth factor (TGF)-beta mediates matrix production in both mesangial and vascular smooth muscle cells. Both TGF-beta and phorbol-12 myristate-13-acetate (PMA) exert both positive and negative effects on mitogenesis in these as well as other cell types. Phorbol esters act through stimulation of protein kinase C (PKC) and are among the most potent tumor promoters known. The present study was conducted to determine if the growth inhibitory effect of TGF-beta parallels that of the phorbol esters and whether this effect of TGF-beta is dependent on activation of PKC. We show that, in vascular smooth muscle cells stimulated to divide by the addition of the serum component basic fibroblast growth factor (bFGF), TGF-beta1 inhibits mitogenesis in a dose-dependent manner, by a maximum of 79% when applied at a concentration of 1 ng/ml. Furthermore, the inhibitory effect on mitogenesis of either TGF-beta1 or PMA, when added four hours after bFGF, are 71% and 84% respectively. Both TGF beta1 and PMA cause translocation of celllular PKC with similar time courses, while neither PKC-alpha nor PKC-betaII are increased in quantity in response to TGF-beta1. In addition, down-regulation of PKC by 24 hours incubation with PMA abolishes TGF-beta's inhibitory effect in bFGF-stimulated cells. We conclude that (i) the signaling pathway utilized by TGF-beta resulting in inhibition of mitogenesis parallels that of PMA, and (ii) the inhibitory effect of TGF-beta1 on bFGF-induced mitogenesis is partially due to activation of PKC. These data suggest that TGF-beta may be an endogenous activator of the growth-inhibitory pathway of PKC, and, since cellular differentiated functions generally occur when the cells are proliferation-inhibited, PKC may be a modulator of extracellular matrix deposition. PMID- 7474660 TI - AVP inhibits EGF-stimulated MAP kinase cascade in Madin-Darby canine kidney cells. AB - We investigated the effects of epidermal growth factor (EGF) and arginine vasopressin (AVP) on Raf-1-MAP kinase cascade, including Raf-1-kinase (Raf-1-K), MAP kinase kinase (MAPKK), MAP kinase (MAPK) and S6 kinase (S6K) in Madin-Darby canine kidney (MDCK) cells. In a dose-dependent manner (10(-10) M to 10(-6) M), EGF increased autophosphorylation of Raf-1-K and activated MAPKK, MAPK and S6K. Sequential activation of these kinases was indicated by their peak times of activation (Raf-1-K 5 min; MAPKK 10 min; MAPK 15 min; and S6K 30 min). AVP (10( 9) M to 10(-6) M) inhibited EGF-stimulated MAP kinase cascade. 8-Bromo-cyclic AMP (cAMP) could mimic the inhibitory effect of AVP on EGF-stimulated MAP kinase cascade. These results were confirmed using H-89, an inhibitor of protein kinase A (PKA) that blocked the effect of AVP on EGF-stimulated MAPK activity. We conclude that AVP inhibits EGF-stimulated Raf-1-K, MAPKK, MAPK, and S6K activity via cAMP in MDCK cells. Our results indicate that MAP kinase cascade may play an important role in integrating the effects of AVP and EGF on distal tubule function. PMID- 7474661 TI - Local macrophage proliferation in the progression of glomerular and tubulointerstitial injury in rat anti-GBM glomerulonephritis. AB - The aim of this study was to examine the contribution of local proliferation in the development of macrophage accumulation and macrophage-mediated injury in rat anti-GBM glomerulonephritis. Using double immunohistochemistry staining of monocyte/macrophages plus the proliferating cell nuclear antigen (PCNA) or bromodeoxyuridine (BrdU) incorporation, we found that the initial accumulation of ED1+ macrophages in the kidney on day 1 of disease was due to an influx of circulating monocytes. However, large numbers of proliferating macrophages (ED1+PCNA+cells), including mitotic macrophages, were present within the glomerulus and interstitium during disease progression (days 7 to 21), accounting for up to 62% of the total macrophage population and giving an excellent correlation with total macrophage accumulation (glomerulus, r = 0.92; interstitium, r = 0.94; both P < 0.001). These proliferating cells had a monocyte phenotype (ED1+ED2-ED3-), but this marked proliferative activity was restricted to the diseased kidney since no PCNA expression or BrdU incorporation was evident within circulating blood monocytes. Proliferating macrophages were almost exclusively localized in areas of severe tissue damage and they correlated significantly with glomerular and tubulointerstitial lesions (P < 0.001), proteinuria (P < 0.001) and creatinine clearance (P < 0.01). In marked contrast, glomerular PCNA- macrophages failed to correlate with these parameters. In conclusion, this study has demonstrated that local macrophage proliferation is the major mechanism of macrophage accumulation during the progression of rat anti GBM glomerulonephritis. Furthermore, it suggests that proliferating macrophages are potent local effector cells in the mediation of progressive renal injury in this disease. PMID- 7474662 TI - Critical subcellular targets of cisplatin and related platinum analogs in rat renal proximal tubule cells. AB - The clinical usefulness of chemotherapeutic agents containing the platinum moiety is often limited by their nephrotoxicity. To investigate the mechanism of nephrotoxicity, and to assess the effects of platinum analogs on specific organelles and basal protein synthesis, biochemical and ultrastructural analyses were performed in rat renal proximal tubule cells (RPTCs). Neutral red (NR) uptake was used to measure lysosomal function, and conversion of MTT to formazan used to assess mitochondrial function. Despite their differential toxicity, cisplatin, carboplatin and CI-973 caused similar progressive inhibition of specific functions, suggesting they may share a common mechanism of nephrotoxicity. Protein synthesis was the earliest indicator of toxicity, followed by NR uptake and MTT conversion. Fluorescent probes for lysosomes (acridine orange) and mitochondria (rhodamine 123) confirmed that cisplatin's toxicity to RPTCs was delayed and cumulative. Condensation of nucleolar components and fragmentation of RER were observed in RPTCs treated for as little as two hours. Since the nucleolus is the site of ribosome biogenesis, the early inhibition of protein synthesis by cisplatin may arise from disruption of this region. In contrast, mitochondrial dysfunction and swelling were late-stage events, and are therefore unlikely to be the primary targets of nephrotoxic platinum compounds. PMID- 7474663 TI - Immunohistochemical detection of advanced glycation end products in dialysis related amyloidosis. AB - beta 2-Microglobulin (beta 2m) isolated from the amyloid deposits in patients with dialysis-related amyloidosis (DRA) has been demonstrated to be modified with advanced glycation end products (AGE). We produced a monoclonal anti-AGE antibody which localized AGE to amyloid deposits in patients with DRA by immunohistochemistry. The connective tissues in the carpal tunnel were obtained from surgical specimens in six patients with DRA. AGE were localized to the beta 2m-positive amyloid deposits in these patients using the monoclonal anti-AGE antibody. AGE were also detected in infiltrating cells surrounding the amyloid deposits. The AGE-positive cells were identified as macrophages, since they showed positive staining with anti-CD68 antibody. In conclusion, AGE were demonstrated by immunohistochemical technique to be present in both beta 2m positive amyloid deposits and surrounding macrophages in patients with DRA. PMID- 7474665 TI - Accuracy of urea removal estimated by kinetic models. AB - The most accurate method for assessing the dialysis dose delivered during high efficiency/flux hemodialysis has not been established. Most current indices of dialysis dose are based on blood-side urea measurements, and thus estimate urea removal. Unfortunately, these methods may lead to inappropriately short dialysis during high flux or high efficiency dialysis, perhaps because of inaccuracies in estimating the amount of urea removal. It is unknown whether these clearance based approaches can accurately predict either absolute or fractional net urea removal, the latter being equivalent to the solute removal index (SRI). Therefore, we compared the urea removal calculated by five blood-side kinetic methods: (1) urea reduction ration, (2) 1-pool, (3) 2-pool models, and the (4) Smye and (5) Daugirdas formulae. These were compared with the gold standard measurement by direct dialysate quantification. Eight stable patients receiving high-flux hemodialysis were studied over four sessions each. BUN was measured at 0, 45 minutes, 90 minutes, end dialysis, one hour after dialysis (equilibrium value), and 48 hours later. Total body water was determined from the dialysate urea removal; the urea generation rate was calculated using one hour post dialysis and 48-hour BUN values. Both the total body water and urea generation rate were provided to the 1- and 2-pool models to optimize accuracy. The urea reduction ratio overestimated SRI. The 1-pool model overestimated both absolute urea removal and SRI in 28 of 32 sessions. The 2-pool model slightly underestimated both absolute urea removal and SRI. In contrast, the Smye and Daugirdas formulas accurately estimated SRI.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7474664 TI - Evidence that reduced lipoprotein lipase activity is not a primary pathogenetic factor for hypertriglyceridemia in renal failure. AB - The aim of the study was to document postheparin plasma lipoprotein lipase (LPL) and hepatic lipase activities and relate these to serum lipid, lipoprotein and apolipoprotein concentrations in 85 patients with kidney function ranging from normal to dialysis dependency. Strict selection criteria were applied in order to exclude conditions other than renal failure which may influence lipid metabolism. Stress was laid on minimizing proteinuria and inflammatory activity. The changes in the lipoprotein pattern were numerically strikingly modest compared to those previously reported. This probably reflected the intention to elucidate the contribution of reduced renal function as such to the dyslipoproteinemia of renal failure, a condition often associated with confounding factors. Significant increases in serum concentrations of triglycerides and apolipoprotein CIII were already observed in moderate renal failure, whereas serum concentrations of high density lipoprotein cholesterol and plasma LPL activities were decreased only in severe renal failure. Plasma LPL activities were not significantly reduced in hemodialysis patients (probably due to anticoagulation with low molecular weight heparin), but serum concentrations of triglycerides and apolipoprotein CIII were significantly increased. A multiple regression analysis, taking glomerular filtration rate, LPL and apolipoprotein CIII into account, showed that both plasma LPL activity and serum apolipoprotein CIII concentration independently predicted serum triglyceride concentration. However, serum apolipoprotein CIII concentration was a much stronger predictor than plasma LPL activity. Thus, a decrease in LPL activity does not seem to be a prerequisite for the hypertriglyceridemia of uremia, but it probably accentuates this condition.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7474666 TI - Antiphospholipids in hemodialysis patients: relationship between lupus anticoagulant and thrombosis. AB - Vascular access thrombosis (VAT) is frequent in some hemodialysis patients. Antiphospholipid antibodies (APL) have been involved in thrombosis, and have been reported to be present in a high proportion of patients with chronic renal failure. We studied the relationship between APL and thrombosis in 97 hemodialysis patients (HD). Lupus anticoagulant (LA) was assessed by activated partial thromboplastin time (APTT) and by tissue thromboplastin inhibition assay (TTI). IgG-anticardiolipin (ACA) was measured by a solid phase ELISA. The prevalence of APL was 31%; LA was found in 16.5% and was detected in all cases by TTI. Only one patient was positive for APTT. ACA was found in 15.5%. Only one patient was positive for LA and ACA. We found no relation between APL and age, length of time on dialysis, sex, type of dialysis membrane, drugs, and chronic B and C hepatitis. A high prevalence of APL was found in patients with undetermined nephropathy. When histories of thrombosis were examined, VAT was found to be significantly more frequent in patients with LA than in patients without LA (62% vs. 26%; P = 0.01). This relation was not present with ACA. Since VAT is one of the most frequent causes of morbidity for HD, diagnostic evaluation of VAT in HD should now include assay for LA. PMID- 7474667 TI - Decreased frequency of HLA-DR13DR6 in Wegener's granulomatosis. AB - The pathogenesis of Wegener's granulomatosis, microscopic polyangiitis and idiopathic rapidly progressive glomerulonephritis (RPGN) is still unclear; in vitro data support both humoral and cellular autoimmune mechanisms. An association of Wegener's granulomatosis with HLA antigens has been described, with conflicting results concerning the antigens involved. We have performed serological HLA typing of patients at two different laboratories within the Netherlands (N = 118 and N = 106,N respectively). A significant decrease in the frequency of HLA-DR13DR6 was present in both patient groups in comparison to controls (chi 2 = 21.9; corrected P value < 0.004 for both groups together). There were no differences in the distribution of HLA-antigens between patients with Wegener's granulomatosis and microscopic polyangiitis, between cANCA (cytoplasmic anti-neutrophil cytoplasmic antibodies) and pANCA (perinuclear ANCA, anti-MPO) positive patients, and between patients with and without relapsing disease. PMID- 7474668 TI - Erythropoietin enhances vascular responsiveness to norepinephrine in renal failure. AB - The mechanism of hypertension induced by recombinant human erythropoietin (rHuEPO) is unclear but may include an increase in peripheral vascular resistance. We studied changes of arterial pressure and plasma endothelin in nine consecutive hemodialysis patients before, and 6 and 12 weeks after, starting rHuEPO. In six patients, changes in cardiac index (CI), stroke index (SI) and total peripheral resistance index (TPRI) were measured by bioimpedance, and forearm vascular responsiveness to intra-arterial norepinephrine (30 to 240 pmol/min) and endothelin-1 (5 pmol/min) were assessed. Six healthy age and sex matched subjects also underwent assessment of forearm vascular responsiveness to norepinephrine and endothelin-1. Treatment with rHuEPO significantly increased hemoglobin and mean arterial pressure (MAP). TPRI also increased by 35 +/- 11%. Plasma endothelin, although elevated basally, remained unchanged. Intra-arterial infusion of norepinephrine caused a maximal increase in forearm vascular resistance (FVR) of 17 +/- 9% before rHuEPO, significantly less than the 32 +/- 5% increase in healthy control subjects (P = 0.04). The response increased to 65 +/- 15% (P = 0.03) after 12 weeks rHuEPO treatment (P = 0.51 vs. controls). Endothelin-1 caused a maximal increase of FVR at 60 minutes of 45 +/- 24% before rHuEPO, which was not significantly different from controls, and tended to decrease with rHuEPO therapy. The response to endothelin-1, but not norepinephrine, correlated inversely with MAP (r = -0.52; P = 0.03) and TPRI (r = -0.51; P = 0.04). In conclusion, these studies show that anemia in chronic renal failure is associated with depressed vascular responsiveness to norepinephrine which is restored by rHuEPO therapy.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7474669 TI - Long-term effects of reduced renal mass in humans. AB - The long-term risks of kidney donation have not been well defined. We carried out meta-analysis of investigations that examined the long-term effects of reduced renal mass in humans. We used multiple linear regression to combine studies and adjust for differences in the duration of follow-up, the reason for reduced renal mass, the type of controls, age and gender. We analyzed 48 studies with 3124 patients and 1703 controls. Unilateral nephrectomy caused a decrement in glomerular filtration rate (-17.1 ml/min; 95% confidence interval -20.2 to -14.0 ml/min) that tended to improve with each 10 years of follow-up (1.4 ml/min/decade; 0.3 to 2.4 ml/min/decade). Patients with single kidneys had small, progressive increases in proteinuria (76 mg/day/decade; 52 to 101 mg/day/decade), but proteinuria was negligible after nephrectomy for trauma or kidney donation. Nephrectomy did not affect the prevalence of hypertension, but there was a small increase in systolic blood pressure (2.4 mm Hg; -0.3 to 5.1 mm Hg, P > 0.05) which rose further with duration of follow-up (1.1 mm Hg/decade; 0.0 to 2.2 mm Hg/decade). Diastolic blood pressure was higher after nephrectomy (3.1 mm Hg; 1.8 to 4.4 mm Hg), but this increment did not change with duration of follow-up. Thus, in normal individuals, unilateral nephrectomy does not cause progressive renal dysfunction, but may be associated with a small increase in blood pressure. PMID- 7474671 TI - Opposite effects of urea on hemoglobin-oxygen affinity in anemia of chronic renal failure. AB - We studied the action of urea on the spin-spin relaxation rate of 2,3 diphosphoglycerate (2,3-DPG) phosphorus atoms in normal and uremic erythrocytes. At concentrations from 10 to 60 mM, urea increased the relaxation rates of 2,3 DPG P-3 phosphorus atoms. This evidenced a stronger binding of 2,3-DPG to hemoglobin (Hb), suggesting that the deoxyform of Hb was stabilized. This hypothesis was confirmed by measurements of the association constant of oxygen to hemoglobin (K) in normal erythrocytes in presence of urea concentrations in the range of those observed in uremic patients (30 mM). Indeed, the observed decrease in K suggests that the T structure of hemoglobin is stabilized. By contrast, with higher urea concentrations (120 mM), measurements of P50 showed an increase in the hemoglobin affinity for oxygen (decrease in P50). Moreover, the relaxation rates of 2,3-DPG P-3 phosphorus atoms were not modified, which is consistent with the simultaneous increase of K. This may be attributed to the formation of carbamylated hemoglobin in presence of urea. These results suggest two opposite effects of urea on Hb-O2 affinity: the first reinforces 2,3-DPG-Hb binding and leads to a decrease in O2 affinity; the second, mediated by carbamylation of Hb, hinders the binding of 2,3-DPG and increases the O2 affinity. These findings are consistent with the fact that, despite the presence of carbamylated hemoglobin, uremic patients do not present increased Hb-O2 affinity. PMID- 7474670 TI - High-resolution B-mode ultrasonography in evaluation of atherosclerosis in uremia. AB - We sought to determine whether artherosclerosis may be accelerated in uremic patients on maintenance hemodialysis and investigated the risk factors for carotid and femoral atherosclerosis in such patients. High-resolution B-mode ultrasonography was used to determine the intima-media thickness (IMT) of the carotid and femoral arteries in 199 hemodialysis patients and 81 age-matched healthy controls subjects. The IMT values of the carotid and femoral arteries in the hemodialysis patients were significantly higher than in age-matched control subjects in most age groups. The IMT values of the carotid or femoral artery were significantly correlated with age in both the hemodialysis patients and the control subjects. There was a significant relationship between the IMT values of the two arteries in the hemodialysis patients (r = 0.418, P = 0.0001) and in the control subjects (r = 0.321, P = 0.0037). Multiple regression analysis showed that age, cigarette smoking, and uremic state were independent risk factors for atherosclerosis of both arteries in the patients and the control subjects (R2 = 0.174, P < 0.0001; R2 = 0.205, P < 0.0001, respectively). In the hemodialysis patients, the independent risk factors associated with the extent of the IMT of the carotid artery were age, cigarette smoking, and serum phosphorus level (R2 = 0.230, P < 0.0001), while those associated with the extent of the IMT of the femoral artery were age, cigarette-smoking, and serum m-PTH level (R2 = 0.230, P < 0.0001).(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7474672 TI - Estimating equilibrated Kt/V from an intradialytic sample: effects of access and cardiopulmonary recirculations. AB - The Smye method has been proposed to estimate the equilibrated post-dialysis BUN based on an additional intradialytic sample obtained approximately one hour into dialysis. However, the effects of access recirculation (AR) and cardiopulmonary recirculation (CPR) on the Smye computation and the corresponding details of how blood is sampled have not been studied. We examined the accuracy of two variations of the Smye technique. In one method, the intradialytic and postdialysis blood samples were obtained at full blood flow. In the other, the samples were obtained after two minutes of slow flow, to obviate the effects of both AR and CPR. Seventeen patients undergoing high efficiency dialysis and three to four-hour treatment times were studied, in whom substantial AR was excluded based on two-minute slow flow urea rebound measurements during and just after dialysis. In this group equilibrated Kt/V (eKt/V) values computed using the Smye derived equilibrated postBUN estimates (full flow samples, 1.22 +/- 0.058 SEM, slow flow samples, 1.23 +/- 0.064) were similar to eKt/V calculated from the 30 minute postdialysis BUN specimen (1.23 +/- 0.049, P = NS). In eight other patients with severe AR (mean 35% +/- 4.5), the accuracy of the full flow Smye estimates was poor when the degree of AR was not constant throughout the dialysis session. Accuracy of the slow flow Smye estimates of eKt/V was unaffected by the presence of severe AR. One advantage of using the full flow Smye method, however, was that a large delta Kt/V value ( < -0.40) was highly suggestive of severe AR.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7474673 TI - Calcium-regulated parathyroid hormone secretion in adynamic renal osteodystrophy. AB - Hypercalcemia and low serum parathyroid hormone (PTH) levels are features of the adynamic lesion (AD) of renal osteodystrophy, but there is little information about parathyroid gland function in this disorder. Therefore, the four parameter model was used to evaluate calcium-regulated PTH release in patients with either adynamic bone or secondary hyperparathyroidism (OF) as documented by bone biopsy and in normal volunteers (NL). Patients had undergone CCPD for 20 +/- 4.2 months, and all received calcium carbonate as the sole phosphate-binding agent. During two hours infusions of sodium citrate, the rate of decline in serum ionized calcium levels did not differ among groups; serum PTH levels rose from 136 +/- 38 to 342 +/- 140 pg/ml in AD and from 691 +/- 99 to 869 +/- 121 pg/ml in OF. Maximum PTH levels were 322 +/- 42% of baseline values in AD but only 146 +/- 9.7% of baseline in OF (P < 0.001), and the increase above baseline levels in AD did not differ from that in NL (300 +/- 25%, NS). During calcium infusions, serum PTH levels fell from 164 +/- 75 to 39 +/- 11 pg/ml in AD and from 622 +/- 76 to 171 +/- 29 pg/ml in OF; minimum serum PTH levels, expressed as a percentage of pre-infusion values, were 25 +/- 2% in AD and 26 +/- 5% in OF (NS).(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7474674 TI - The PiZ gene of alpha 1-antitrypsin as a determinant of outcome in PR3-ANCA positive vasculitis. AB - We have previously demonstrated that a strong correlation exists between systemic vasculitis with proteinase 3-anti-neutrophil cytoplasm antibodies (PR3-ANCA) and heterozygosity for alpha 1-antitrypsin (alpha 1-AT) deficiency, PiZ. In the present study we characterized the PiZ-positive subgroup by laboratory findings, clinical features and outcome. The series studied consisted of 18 PiZ-positive and 81 PiZ-negative PR3-ANCA patients, comparable in sex ratio, age, C-reactive protein concentrations and renal function at diagnosis, and treatment. PiZ positive patients had a more disseminated disease as reflected by the number of affected organs (P < 0.01). We found no group differences in relapse tendency. Overall mortality was 39% (7 of 18) in the PiZ-positive and 16% (13 of 81) in the non-PiZ group (P = 0.048). When survival analysis was restricted to 66 patients included in the study at disease onset, the group difference was significant (P = 0.016). The results suggest that the subnormal response of plasma alpha 1-AT seen in PiZ-heterozygotes enhances the risk of dissemination of the vasculitic process and the risk of a fatal outcome. We consider alpha 1-AT phenotyping to be justified in cases of PR3-ANCA-positive vasculitis. Treatments decreasing plasma alpha 1-AT (such as plasmapheresis without plasma replacement) may be deleterious. PMID- 7474675 TI - "Strict" blood pressure control and progression of renal disease in hypertensive nephrosclerosis. AB - Hypertensive nephrosclerosis is a progressive renal disease and the leading cause of end-stage renal disease (ESRD) in blacks in the United States. It is generally believed that hypertensive renal injury is responsible for progressive renal failure; however, it is not known whether pharmacologic lowering of blood pressure to any level prevents progression of renal disease. Accordingly, we performed a long-term prospective randomized trial to determine whether "strict" [diastolic blood pressure (DBP) 65 to 80 mm Hg] versus "conventional" (DBP 85 to 95 mm Hg) blood pressure control is associated with a slower rate of decline in glomerular filtration rate. Eighty-seven non-diabetic patients (age 25 to 73; 68 black, 58 male) with long-standing hypertension (DBP > or = 95 mm Hg), chronic renal insufficiency (GFR < or = 70 m/min/1.73 m2) and a normal urine sediment were studied. DBP was pharmacologically lowered to < or = 80 mm Hg (3 of 4 consecutive measurements at 1 to 4 weeks intervals) after which patients were randomized. DBP and GFR (renal clearance of 125I-iothalamate) were measured at baseline, at three months and every six months post-randomization. The rate of decline in GFR (GFR slope, in ml/min/1.73 m2/year), estimated by the method of maximum likelihood in a mixed effects model, was the primary outcome variable. In a secondary analysis, 50% reduction in GFR (or a doubling of serum creatinine) from baseline, ESRD and death were combined.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7474676 TI - Naloxone inhibits renal hemodynamic effect of head-out water immersion in humans. AB - Head-out water immersion (HOI) is followed by renal vasodilation and natriuresis, in association with a fall in blood pressure. The latter suggests an exaggerated sympathetic suppression. We studied the role of endogenous opioids on the renal response to HOI. Six healthy subjects underwent four- hour clearance studies during: (1) time control; (2) naloxone 0.1 mg/kg i.v. bolus, followed by 0.1 mg/kg/hr; (3) HOI; and (4) concomitant HOI and naloxone administration. Compared to the time control study, naloxone had no effects on mean arterial pressure (MAP), plasma renin activity (PRA), aldosterone, catecholamines, atrial natriuretic peptide (ANP), glomerular filtration rate (GFR), estimated renal plasma flow (ERPF), and sodium excretion. HOI caused significant decrements of MAP, PRA, aldosterone, and catecholamines, and increased ANP, GFR, from 94 +/- 5 to 102 +/- 5 ml/min (P < 0.01), ERPF, from 529 +/- 30 to 616 +/- 35 ml/min (P < 0.01), and sodium excretion. Renal blood flow increased as well, and calculated renal vascular resistance decreased from 99 +/- 6 to 77 +/- 5 mm Hg.min.liter-1 (P < 0.01). HOI during concomitant naloxone administration had similar effects on MAP and humoral factors, however, caused no change in GFR, ERPF and renal blood flow, and the fall in renal vascular resistance, from 98 +/- 6 to 83 +/- 5 mm Hg.min.liter-1 (P < 0.05) was significantly less than found in the absence of naloxone (P < 0.05). The natriuretic effect was undisturbed. These data suggest that endogenous opioids play a role in the response to HOI, in particular, potentiate the renal vasodilatory response. PMID- 7474678 TI - The chemistry of urinary stones around 1800: a first in clinical chemistry. AB - At the end of the 18th century, as soon as modern chemistry was created, dedicated physicians tried to apply it to medicine. A rewarding field was that of urinary lithiasis. Stones offered a sufficient amount of a relatively pure chemical present in the body. Indeed, urine and the solidified matter was within the analytical grasp of the existing techniques. The first step was made by Scheele in Sweden who identified uric acid in calculi and normal human urine. During the following thirty years, Fourcroy and Vauquelin in Paris and Wollaston, Pearson, Marcet and Prout in London identified the various salts, uric acid, urate, various phosphates, oxalate, calcium, ammonium and magnesium forming current calculi. The conditions of their solubility in vitro were described. Even rare components such as cystine and xanthine were unraveled in London. The impetus given by these studies offered a good start to clinical chemistry in general and to the understanding of urinary lithiasis in particular. This led to the discovery of the corresponding solutes in the urine of such patients. Unfortunately, during the following decades nothing was added to these chemical investigations, and most of what had been acquired was forgotten. Research was concentrated on clinical pathology. The implementation of chemistry to medicine had to wait. PMID- 7474677 TI - The standard peritoneal permeability analysis: a tool for the assessment of peritoneal permeability characteristics in CAPD patients. AB - Peritoneal transport characteristics in CAPD patients are often assessed by the peritoneal equilibration test (PET), which uses a four hour dwell with glucose 2.27% dialysate. From the test, the dialysate/plasma ratio of creatinine (D/PCr), the dialysate/initial dialysate ratio of glucose (D/Do) and net ultrafiltration (NUF, drained minus instilled volume) are calculated. The standard peritoneal permeability analysis (SPA) is a modification and extension of the PET: glucose 1.36% dialysate is used, to which dextran 70 (1 g/liter) is added for the calculation of fluid kinetics. Mass transfer area coefficients (MTAC's) of low molecular weight solutes, clearances of proteins and the change in intraperitoneal volume (delta IPV) can be assessed. In this study the SPA was analyzed, and a comparison with the PET was made. A total number of 138 SPA's was analyzed in 86 different clinically stable patients. Normal values were calculated for both SPA and PET parameters in the same tests. Median (ranges) of comparable transport parameters from SPA and PET were: MTACCr, 10.4 ml/min (5.7 to 19.3); glucose absorption, 61% (35 to 87); delta IPV, 9.5 ml (-761 to 310); D/PCr, 0.76 (0.53 to 1.14); D/D0, 0.37 (0.13 to 0.56); NUF, -75 ml (-675 to 450). The agreement between SPA and PET was analyzed using the method of Bland and Altman. A fairly good agreement was present between NUF and delta IPV. Systematic errors were found when D/PCr and MTACCr were compared: D/P overestimated MTAC mainly in the low range, whereas in the high range values were underestimated. A similar pattern was seen for the transport parameters of glucose. In 40 patients negative net ultrafiltration was present, and possible reasons for this were assessed. In 9 patients no reason could be identified. It can be concluded that the SPA provides useful and extensive information on peritoneal transport parameters. Compared to the PET, the SPA has better discriminative power for the transport of glucose and creatinine. PMID- 7474680 TI - Pathogenesis of diabetic nephropathy: experimental approaches. The Second Hyonam Kidney Laboratory International Symposium. Seoul, Korea, January 21, 1995. Proceedings. PMID- 7474679 TI - Hantavirus nephropathy. PMID- 7474681 TI - Effects of high glucose concentration on c-fos: role of protein kinase C. PMID- 7474682 TI - Role of oxidative stress in the development of diabetic nephropathy. PMID- 7474683 TI - Effect of high glucose concentration on phospholipase A2 activity in mesangial cells. PMID- 7474684 TI - Calcium signaling and contractile response of diabetic glomerular mesangial cells. PMID- 7474685 TI - High glucose activates protein kinase C and stimulates fibronectin gene expression by enhancing a cAMP response element. PMID- 7474686 TI - Role of transforming growth factor-beta in diabetic glomerulosclerosis and renal hypertrophy. PMID- 7474687 TI - Growth hormone expression is required for the development of diabetic glomerulosclerosis in mice. PMID- 7474688 TI - Effect of glucose and cytokines on the expression of cell adhesion molecules on mesangial cells. PMID- 7474689 TI - Advanced glycation in diabetic renal and vascular disease. PMID- 7474690 TI - Endothelins in diabetic kidneys. PMID- 7474691 TI - Does nitric oxide, an inflammatory mediator of glomerular mesangial cells, have a role in diabetic nephropathy? PMID- 7474692 TI - Effect of HMG CoA reductase inhibition on TGF-beta 1 mRNA expression in diabetic rat glomeruli. PMID- 7474693 TI - [Difficulties, errors and complications in laparoscopic cholecystectomy]. AB - The work deals with the specific features of surgical techniques in laparoscopic cholecystectomy. It discusses in detail the most frequently encountered difficulties, errors, and complications developing at various stages of the operation. From their own experience the authors describe the causes and mechanisms of their occurrence. A greater part of the article is concerned with rational surgical tactics in a complicated course of laparoscopic cholecystectomy. It is emphasized that the indications and contraindications for laparoscopic cholecystectomy must be strictly observed, all stages of the intervention must be technically carried out with great care and scrupulously, the surgeon must evaluate adequately his own possibilities, sufficient technical equipment must be provided. PMID- 7474694 TI - [Acute cholecystitis as a problem of laparoscopic surgery]. AB - The article deals with experience in 89 operations of laparoscopic cholecystectomy in patients with acute cholecystitis aged 17 to 82 years. A total of 92 attempts were made, 3 patients underwent laparotomy. Forty-five patients had catarrhal, 35 phlegmonous, and 9 gangrenous cholecystitis. In the preoperative period 37 patients were subjected to endoscopic retrograde pancreaticocholangiography, endoscopic papillosphincterotomy was performed in 28 of them, in 9 the operation was complemented by nasobiliary drainage. As a result intraoperative cholangiography had to be conducted only in 3 patients. Intraoperative complications developed in 42 and postoperative in 18 patients (local in 15 and general in 3). There were no fatal outcomes. PMID- 7474695 TI - [Technical principles of laparoscopic cholecystectomy]. AB - The technical principles of laparoscopic cholecystectomy are described from experience in 87 operations in acute and chronic appendicitis. The authors discuss the stages of the operation and the peculiarities of the technical procedures at each stage, and give recommendations on the use of the instruments and indications for abdominal drainage. PMID- 7474696 TI - [Experience in 735 cases of laparoscopic cholecystectomy]. AB - In the period from December, 1991 to December, 1993, 735 successful operations of laparoscopic cholecystectomy (LC) were conducted in the III Surgical Clinic of the Semmelweis Medical University, Budapest, Hungary. Among the patients were 154 females and 581 males 14 to 82 years of ago. Conversion of laparotomy was undertaken in 58 (7.9%) patients because of technical difficulties and the development of complications. Intraoperative complications were encountered in 197 (26.8%) and postoperative complications in 38 (5.17%) patients. PMID- 7474697 TI - [Failures, errors and complications of laparoscopic cholecystectomy]. AB - Failures and complications are analysed from experience in 325 operations of laparoscopic cholecystectomy. Change to laparotomy was undertaken in 5 (1.5%) patients. The total amount of complications was 45.2%, among them 36.5% were intraoperative and 8.7% postoperative, but only 3.8% were hazardous for the patients. Mortality was 0.3% (one patient dies from cardiovascular failure). A classification of complications and the technical principles of the operation are discussed. PMID- 7474698 TI - [Immediate, early and late results of endoscopic papillosphincterotomy]. AB - Clinico-instrumental examination of 226 patients showed that there were no fatal outcomes directly after endoscopic papillosphincterotomy, the number of complications was approximately the same as after transduodenal operations on the major duodenal papilla but they took a much easier course. The high effectiveness of an endoscopic operation was confirmed in the immediate postoperative period (absence of biliary hypertension, jaundice, pancreatitis, cholangitis). In the late-term periods after EPST restenosis was rarely encountered (3.9%); duodeno biliary reflux caused no serious disorders, while the functional activity of the sphincter Oddi which had been operated on was partly maintained in 84.2% of patients. PMID- 7474699 TI - [Role and perspectives of the use of laparoscopic surgery in colo-proctology]. AB - A comparative analysis of the performance of 51 operations for resection of various parts of the colon, among which were 24 operations conducted by laparoscopy, was undertaken. Certain advantages of laparoscopic resection of the colon, as well as the possibility of lymph dissection, which meets the principles of oncological surgery, were revealed. PMID- 7474700 TI - [Immediate results of laparoscopic hernioplasty in inguinal and femoral hernias]. AB - In 1992-1994 laparoscopic hernioplasty was performed in 403 patients with inguinal (371) and femoral (32) hernias. Mean duration of hospital-stay 2.3 days, period of incapacity 9.7 days. Complication were encountered in 25 (3.7%) patients, including perforation of the small intestine and commissural ileus for which laparotomy had to be performed. Recurrent hernias occurred in 7 (1.7%) patients in periods of 2 weeks to 1.5 month due to poor fixation of the implant and choice of an inadequate operative method. PMID- 7474701 TI - [Video thoracoscopic sympathectomy]. AB - Twenty operations of video thoracoscopic sympathectomy (VTS) were performed on 17 patients in 1992-1994. The indications for the operation are Raynaud's disease and some obliterating occlusions of the upper limb arteries in which a reconstructive operation is not possible. The VTS operation is technically simple and atraumatic. Its effectiveness is determined by the disappearance of pain, warmer hands, and the results of histological study. The authors experience has shown that VTS does not need special instruments. At the same time, they believe that cooperation with specialists in thoracic and vascular surgery is very important and obligatory in development of complications. PMID- 7474702 TI - [Spectroscopic diagnosis of stomach diseases based on laser-induced fluorescence of endogenous porphyrins]. AB - For higher efficacy of early gastric carcinoma diagnosis by endoscopy, the authors suggest a method of autofluorescent diagnostics. It is d=based on the property of endogenous porphyrins contained in the tumor to emit light under the effect of laser. The suggested method was used in examination of 54 patients. The experimental group was composed of 35 patients with gastric carcinoma, the control group was formed of 21 patients with benign diseases gastric ulcer, polyps, chronic gastritis). The sensitivity of the method was 94%, th specificity 90.5%. It is shown that the method of autofluorescent diagnostics may be applied in cases which are difficult for diagnosis. PMID- 7474703 TI - [Clinical fluorescent diagnosis of tumors using photosensitizer photogem]. AB - During clinical trials of photodynamic therapy (PDT) the tissue fluorescence spectra under 510 nm laser excitation were recorded in 22 patients with tumors of the lungs, larynx, skin, gastric and esophageal carcinoma, and cancer of the gynecological organs before (autofluorescence) and after drug administration. A spectroscopic fluorescence detection system, developed at the General Physics Institute, Russian Academy of Sciences, was applied for tumor diagnostics. The system has been used for clinical fluorescence diagnostics with the aid of HPD type photosensitizer Photogem (Moscow Institute of High Chemical Technologies) marked by selective retention in malignant tumors. Methods and equipment for spectra analysis aimed at estimating drug distribution in the near-surface layers of tissues, determining the tumor boundaries before photodynamic treatment, as well as studying drug pharmaco kinetics have been developed and tested. The results demonstrate the possibility of tumor detection by fluorescence even in low selectivity of drug accumulation which appears to be dependent on the stage and type of the disease and the organ involved. PMID- 7474704 TI - [Lymph node dissection in patients with cancer of the proximal part of the stomach]. AB - The authors describe 694 patients who underwent operation in 1985-1993 for carcinoma of the proximal part of the stomach with various degree of involvement of the adjacent parts of the stomach and esophagus. The tumors differed in histological structure. The main principles of classification of the groups of regional lymph nodes, taking into account the localization of the tumor in the stomach, are shown. These Principles are applied by Japanese surgeons in performing lymph node dissection and were modified in the Clinic od Thoracic Oncology of the Scientific Research Institute, Oncological Center, Russian Academy of medical Sciences. The localization and incidence of metastases in disseminated carcinoma of the proximal stomach with involvement of the esophagus were studied. The sequence of the stages of intraabdominal lymph node dissection in proximal gastric resection and gastrectomy is discussed. The tendency towards improvement on the indices of 3-year survival after expanded operations is noted. Expanded operations with scrupulous morphologic examination of the removed lymph nodes is very important for more precise determination of the stage and individual prognostication of the choice of the postoperative methods of treatment. In palliative interventions the operation is cytoreductive in essence and may create to a certain measure better conditions for chemotherapy. PMID- 7474705 TI - [Precardial proximal vagotomy with pyloroplasty in surgery of chronic duodenal ulcers]. AB - The performance of a new operation, precardial proximal vagotomy, for treatment of chronic duodenal ulcer is substantiated, described, and its results are discussed. It consists in division of the proximal vagal branches, stretching to the gastric acid-producing zones, at the level of the cardia, without cutting the blood vessels and sympathetic fibres. This operation in combination with pyloroplasty and temporary drug blockade of the celiac plexus excels in efficacy all the other methods of surgical management of duodenal ulcer. According to wide indications, it was conducted on 149 patients, in 68 of them in combination with the above-named additional interventions. During a 7-year follow-up period of the late results, a recurrent ulcer was encountered in one patient and group III visick trouble in another one. PMID- 7474706 TI - [Clinical-functional classification of combined occlusive lesions of the arterial system]. AB - For a complex approach to the clinico-functional appraisal of the vascular system the authors suggest dividing the arterial system into 4 conventional functional basins: the coronary region (1), the brachiocephalic (2), the visceral (3), and the arterial basin of the lower extremities. In accordance with the 4 distinguished anatomo-functional vascular basins, the classification is designated by their first letters: HBVL, where H stands for the heart, B for the brain, V for the visceral vessels, and L for the lower extremities. Next to each letter is shown the functional class of arterial insufficiency of the vascular basin. The figure 0 denotes the absence of arterial stenosis. All the other figures from 1 to 4 reflect the functional severity of arterial insufficiency. The 1st functional class is characterized by the absence of a clinical picture of the disease in arterial stenosis, the 2nd class by the stage of subcompensation of circulation, and the 3rd and 4th classes by decompensation of circulation. The criteria for separating the degree of ischemia into functional classes are suggested. The HBVL classification is a working one because it not only illustrates and indicates fully the condition of arterial circulation in the vessels of the region but determines the tactics of treatment of the patient. The possibilities of the classification are illustrated by concrete clinical cases. PMID- 7474707 TI - [Characteristics of interrelations of external respiration, gas exchange and hemodynamics in patients with bronchopulmonary diseases]. AB - In 60 patients with stage II-III bronchogenic carcinoma of the lung the following procedures were included in the complex of preoperative functional examination: spiro-body-plethysmography, catheterization of the right heart, and simultaneous examination of gas exchange using a DELTATRAC MBM-100 metabolic monitor. To evaluate the lung diffusion capacity and the dimensions of the arteriovenous shunt, oxygen mixtures differing in O2 concentration were used. The results were interpreted within the framework of a homogeneous model of lung gas exchange realized on a PC/AT personal computer. The interrelationship of ventilation disorders with hemodynamic and lung gas--exchange function was studied. Arterial hypoxemia was mainly observed in obstructive and mixed disorders of ventilation function and was more marked in the latter. Primary disturbance of the lung diffusion capacity was found to be the leading link in the development of acute respiratory insufficiency after the operation. PMID- 7474708 TI - [Electric stimulation of the spinal cord in diffuse occlusive lesions of peripheral arteries]. PMID- 7474709 TI - [Plastic surgery in stenoses and intraoperative injuries of the urinary tract]. AB - The author showed the practical significance and expediency of performing reconstructive-plastic operations (57 patients) in surgical correction of severe pathology of the urinary tract. The indications for plastic operations were stenoses of various etiology, intraoperative traumas of the urinary tract, contracted bladder and the consequent ureterohydronephrosis, renal insufficiency. The choice of the tactics and method of surgical treatment should be strictly substantiated and based on the individual features of the urinary system pathology, the character of the intraoperative trauma of the ureter and bladder and its possible after-effect on the kidney. Taking into account the variability of the pathology and the length of the cicatricial narrowing of the pelvic ureteral segment and ureter, and the degree of kidney constriction, variations of plastic operations differing in the complexity of the techniques were applied. These were small in volume flap plastics of the pelvic-ureteral segment and ureter (Calp-de-Virda, Baori, Demela) and intestinal plastics of the bladder in one of the modifications. The possibility of plastic operations in restoration of physiological urination and social rehabilitation of patients with urinary fistulas was determined. According to early and late-term postoperative follow up, the results of the operation were favorable in 91.3% of cases. Postoperative mortality in different periods (7 days to 3 years) was 8.7%. PMID- 7474710 TI - [Injuries of the ducts in surgery of the biliary tract]. AB - The authors analysed 92 reconstructive and restorative operations on the biliary tract injured at a high level during emergency operations. Such complications occur most frequently when the surgeon is insufficiently experienced or due to unavailability or incomplete use of the radiological method of intraoperative diagnostics. The expediency of reinfusion of the lost bile into the gastrointestinal tract in the diagnosis of injury to the biliary tract is pointed out. It is emphasized that patients must be transferred early to a specialized medical establishment for a reconstructive operation. The creation of hepato digestive anastomoses on a transhepatic drain produces better results than the other drainage methods. In the group of 92 patients 77 recovered and 15 died in different periods after the operation. PMID- 7474711 TI - [Dynamic control of acute crisis of rejection of the transplanted heart based on echocardiography]. PMID- 7474712 TI - [Abdominal endoscopic surgery: brief history and present-day state]. PMID- 7474713 TI - [Acute appendicitis in a patient with fibroma of the cecum]. PMID- 7474714 TI - [Polypoid carcinoids of the stomach and rectum]. PMID- 7474715 TI - [First experience with laparoscopic vagotomy in the treatment of peptic ulcer]. PMID- 7474716 TI - [The transurethral use of laser therapy in urology]. AB - The utilization of laser energy plays an ever increasing role in medicine, surgical practice in particular. This is most clearly expressed in the field of urology where endoscopic access to the entire urogenital system is possible against the background of a continuous improvement of the surgical armamentarium over the past few years. The various types of lasers available, qualities of the laser source, and the physical and biophysical principles of their action are comprehensively described. The creation of this particular type of energy (laser), transmissible along flexible quartz fibers and not absorbed by water, considerably increases the interest in laser treatment of diseases of the urinary bladder and urethra. Laser therapy has important clinical implications, and broadens the scope of endoscopic treatment of a variety of urological diseases involving the upper urinary ways. PMID- 7474717 TI - [The organization and function of donor transplantation units]. AB - The donor unit are organized in hospital centers for brain death patients. They function as emergency centers with the possibility to admit patients presenting multiple injuries and neurosurgical problems. Multidisciplinary teams aided by the local coordinator take part in the process of explantation and transplantation. Emphasis is laid on the implantation role played by anesthetist resuscitator, surgeon and local coordinator. PMID- 7474718 TI - [The diagnosis, biopsy and treatment follow-up by ultrasound in patients with prostatic carcinoma]. AB - Prostate carcinoma accounts for 1-5 per cent of all neoplasms affecting men, and represents 29.2 per cent of the tumors involving the urogenital system. It is a disease encountered in advanced age mainly--between 60 and 70 years, and recently even earlier. It is pointed out that transrectal ultrasound examination is usually used for diagnosing prostate carcinoma, determination of its stage, and follow-up study of the treatment effect. The clinical relevance of transrectal echography augments whenever performed in conjunction with biopsy of the prostate under ultrasound control, especially in the event of impalpable neoformation, or in uncertain negative, digitally oriented biopsies. The screening method proposed is capable to provide for a sufficiently accurate orientation towards the site for performing control biopsy of the prostate, but it could hardly serve as a reliable, independent criterion for estimating the effect of the nonoperative treatment in course. PMID- 7474719 TI - [Subvesical obstruction following chronic bacterial prostatitis]. AB - Twenty-eight men below 50 years of age, presenting obstructive miction complaints subsequent to chronic bacterial prostatitis cure, are subjected to operation and follow-up study over a 6-year period. The test of Meares (1991) is used as a diagnostic method to rule out chronic bacterial prostatitis. The patients are distributed according to microbiological findings in the ejaculate in the stage of prostatitis treatment. Obstruction of the lower urinary tracts is diagnosed on the ground of past history data, uroflowmetry, residual urine assessment, miction cystourethrography and urethrocystoscopy under regional anesthesia. In nine cases (32.1 per cent) transurethral resection of the neck of the urinary bladder is performed, and in the remainder (19 cases or 67.9 per cent)--transurethral incision of the cervix vesicae at 5-7 hours. In the postoperative period, 75-89 per cent of patients are free of any miction complaints. PMID- 7474720 TI - [A comparative assessment between transrectal echography, venous urography, CAT and morphology in patients with prostatic carcinoma]. AB - By incidence rate prostate gland carcinoma ranks second among all neoplastic diseases. As all tumorous neoformations, it is conspicuous for its progressive character. Clinical, laboratory and roentgen methods, thus far routinely used in practice, are hardly in a position to establish correct and accurate diagnosis alone. The diagnostic potentials of transrectal ultrasound examination, intravenous urography, CAT and morphological patterns are comparatively studied in patients presenting prostate gland carcinoma. The obtained results point to a greater diagnostic relevance of transrectal ultrasound examination, by comparison with CAT and intravenous urography. It is a matter of a noninvasive and highly effective procedure for primary diagnosis of the neoplastic process. Emphasis is laid on the necessity to confirm the echographic diagnosis in all instances by performing morphological study in conjunction with the remaining diagnostic methods. PMID- 7474721 TI - [Endovesical surgery via the suprapubic approach]. AB - Experience had with suprapubic cystoscopy and endovesical surgery, performed in a series of twenty-two patients presenting various diseases of the bladder--tumors, mainly of the anterior wall, calculi, foreign bodies, bladder neck sclerosis, defective Foley catheter--is shared. As a sole complication a case of penetrating the abdominal cavity without inflicting lesion to the intestine, is described. The suprapubic approach significantly broadens the scope of transurethral surgery, preserves its advantages, and is furthermore free of the shortcomings associated with open operations. PMID- 7474722 TI - [The C-reactive protein level of patients after the extracorporeal lithotripsy of urinary calculi]. AB - Within 24 to 48 hours of sustaining a variety of severe clinical conditions, such as general bacterial infection, tissue necrosis, acute rheumatism, myocardial infarction, active pulmonary tuberculosis and neoplastic processes, the serum level of C-reactive protein may show a considerable increase. It may also serve as a criterion for the evolution of the disease, and for the effect of the therapy in course. The task undertaken is to use the serum level of C-reactive protein as a criterion for evaluating the extent of renal parenchyma damage following extracorporeal lithotripsy of urinary calculi (ELUC). Proceeding from the results of the study, it is assumed that ELUC is a nontraumatic operative intervention for the renal parenchyma. In case of extreme increase in C-reactive protein level, a higher incidence rate of post-ELUC complications may be anticipated. PMID- 7474723 TI - [The vascular aspect of kidney transplantation]. AB - The vascular aspect of renal transplantation is a factor of major importance throughout the various phases of surgery. It includes nephrectomy from living donors, or intact cadaveric kidney with a sufficiently long renal artery and vein (with or without patch), organic ex situ surgery, and implantation surgical technique. In 1994 fifteen renal transplantations are performed in the Clinical Center of Urology. In thirteen patients the kidneys transplanted are obtained from living donors, and in two-from cadavers. The vascular surgery techniques used, and the problems faced during their application are surveyed. Vascular surgery performed successfully is among the factors guaranteeing good survival of the organ transplanted. PMID- 7474724 TI - [Benign neoplasms of the urethra in women]. AB - This is a report on the treatment of urethral neoplasms in a series of 331 with ages ranging from 21 to 88 years. The most common symptoms of manifestation of the lesion include: bleeding, burning and stinging during urination, eventually pain associated with movements. In forty-one patients the polyps are removed operatively through excision, in 185-through electrocoagulation, and in eighteen excision with ensuing electrocoagulation Eighty-seven patient are subjected to conservative treatment because of the presence of urethral mucosa prolapse. The histological findings show evidence of polyps of the urethra in 206 cases, and urethral caruncle in thirty-six. The obtained results are estimated as good. Mass screening examinations of women are recommended. PMID- 7474725 TI - [Vascular complications in kidney transplantation (a review of the literature and the authors' own experience)]. AB - Vascular complications following renal transplantation are by no means rare. Usually they are multifactorial in nature, and eventually require operative management. Arterial and venous thromboses, arterial stenosis and hemorrhages are among the most frequently encountered vascular complications. In 1994, in the renal transplantation department of the Clinical Center of Urology three patients presenting vascular complications after renal transplantation are observed and treated. Two of them undergo surgery: resection of the stenotic segment of arteria renalis is done in one of them, and explantation-in the second. In the third case it is a matter of tense terminoterminal anastomosis due to technical reasons with ensuing venous thrombosis-here no operative correction is considered owing to intercurrent ischemic attack. In patients with transplantation the timely made diagnosis of vascular complications, and their surgical management prolongs the survival term. PMID- 7474726 TI - [The laparoscopic plastic repair of inguinal hernias]. AB - Laparoscopic plasty of inguinal hernia is an operation performed for the first time in this country. The results of ten operated hernias in 8 patients are analyzed. The operative intervention consists in the application of a special polypropylene sheet over the direct and indirect opening of the inguinal canal, attached with a clasping instrument. The peritoneum is dissected free in advance, and thereafter used for covering the sheet. No complications are observed. PMID- 7474727 TI - [Current surgery of the nasal septum]. AB - Rhinoseptoplasty, septoplasty in particular, is among the most commonly used operations in otorhinolaryngology. So far, over a long period of time, a variety of operative procedures successful for their time have been developed. The present report deals with a total of 4700 patients, operated on over an 18-year period (1977 through 1994). Pertinent physiological and anatomical data concerning the nasal septum are briefly outlined. A number of operative methods are analyzed. Proceeding from the postoperative results, preference is given to Cottle's procedure. PMID- 7474728 TI - [A case of an impossible cannulation of the v. jugularis and v. subclavia in a female patient with longstanding myasthenia gravis and immunosuppressive therapy]. AB - The report deals with a female patient presenting myasthenia gravis of 8-year duration, maintained on continuous APV, repeatedly cannulated central veins, and immunosuppressive therapy with corticosteroids--Imuran and Sandimmun. Repeated CT examination of the mediastinum fails to detect any process. The consecutive scanning is necessitated because of the impossibility to cannulate v. jugularis and v. subclavia at the last attempt. Cytological evidence of a thymosarcoma is established during examination. PMID- 7474729 TI - [A case of generalized convulsion during induction with Diprivan]. AB - The case report concerns a male patient presenting oligophrenia and epilepsy of long duration, adequately prepared for anesthesia with anticonvulsants, sustaining generalized convulsion during anesthesia induction with Diprivan. On deepening Diprivan anesthesia, the convulsion subsides. The patient runs an uneventful postoperative course, free of convulsions. The issue of small doses Diprivan giving rise to convulsions is comprehensively discussed. PMID- 7474730 TI - [Accidentally discovered kidney carcinoma during autopsy]. PMID- 7474731 TI - [An instrument for placing ligatures during liver resection]. PMID- 7474732 TI - [The characteristics of ureteral lithiasis]. AB - The study covers 2630 patients with ureteral concrements, eliminated operatively, through extracorporeal lithotripsy (ESWL), or spontaneously as the result of drug treatment. Attention is called to the left-side location of ureteral concrements, regardless of the mode of elimination, with the clinical symptomatology of the disease being comprehensively discussed. Clinical and chemical characterization of ureteral concrements (type, size and the like) is also done separately with a special reference to the method of removal employed. In concrements eliminated spontaneously a correlative dependence with the annual season is sought; in the winter months a rather elevated frequency of elimination is recorded, irrespective of the type of concrements. PMID- 7474734 TI - [Nonparasitic liver abscesses]. AB - Abscess of the liver is a comparatively rate, but very serious disease, necessitating the undertaking of diagnostic and therapeutic measures on an emergency basis. The principal diagnostic method currently in use includes two imaging techniques--echography and CT. In therapeutic respect the target puncture under echographic or CT control has rather limited scope of application. Surgical therapeutic approach is the leading one--laparotomy, drainage of the abscess, and arbitrary use of a Kerr drainage for the choledochus and regional antibiotic therapy. PMID- 7474733 TI - [The restoration of urinary tract continuity in kidney transplantation]. AB - In 1994, fourteen kidney transplantations are done in the Clinical Center of Urology--FAH over a 4-month period. The patients are with age ranging from 11 to 44 years. In twelve cases the kidneys are obtained from living related donors, and in the remainder--from cadavers. The surgical techniques used to restore urinary tract continuity include: ureteroneocystostomy--in 7 cases, pyeloureterostomy--six, and ureteroureterostomy--one. The choice of a surgical method in transplantation depends on the state of the urinary tract. Assessment of the efficacy of these surgical procedures is practicable after long-term follow-up with a view to eventual late urological complications development. PMID- 7474735 TI - [Our experience with treating acute noncalculous cholecystitis]. AB - Based on a comprehensive literature survey, personal experience with the management of 89 patients, presenting acute noncalculous cholecystitis, is shared. All patients are clinically observed in the acute phase of the disease. In 60 cases the latter becomes manifest with recurrent attacks, and in 29 there are no previous signs or symptoms of biliary diseases. Diagnosis is made on the ground of active echographic and clinical observation. Conservative therapy is effective in 28 cases. The remainder (61) are operated at 1 to 4-day intervals after the onset of the disease because of persisting symptomatology and/or deterioration of the patient's condition. During laparotomy the gallbladder presents marked enlargement with thickened, exudate-imbibed wall. Cholecystectomy is done in all 61 cases. In twelve of them, owing to dilatation of the common bile duct and minimal serum bilirubin increase, stenosing papillitis is diagnosed by intraoperative cholangiography, treated by external biliary drainage and endoscopic sphincterotomy, undertaken after improvement of the general condition. Acute inflammation, free of fibrosis, with isolated foci of gangrene and necrosis of the bladder wall are histologically documented. In four senile patients presenting serious concomitant diseases, with exceptionally high operative risk, echographic percutaneous transhepatic cholecystectomy is resorted to, followed by antibiotic bladder instillations with a very good outcome. The summed up results of acute noncalculous cholecystitis treatment are estimated as very good. PMID- 7474736 TI - [The treatment of acute cholecystitis complicated by mechanical jaundice]. AB - After analyzing the summed up treatment results in 368 patients with acute cholecystitis, complicated by obstructive jaundice, the inference is reached that it is a matter of a complication, unfavourable in terms of prognosis, with a definite impact on both immediate and long-term therapeutic results. Early operative intervention following intensive preoperative preparation (including ERHPG with endoscopic papillosphincterotomy and ensuing biliary decompression, or echographic percutaneous cholecystectomy) contribute to control the acute inflammatory process in the gallbladder, and preclude the development of liver (or hepato-renal) failure against the background of obstructive jaundice. Proceeding from the severe, irreversible changes in liver and kidney, hardly responding to treatment, and persisting deterioration of the general condition, acute cholecystitis complicated by obstructive jaundice is defined as an emergency life-endangering condition, requiring a priority operative approach. PMID- 7474737 TI - [Longmayr's operation in malignant jaundice with a poor prognosis]. PMID- 7474738 TI - [Hallstedt's choledochostomy]. PMID- 7474739 TI - [A new method for the surgical treatment of hepatic echinococcosis]. AB - A procedure for operative management of liver echinococcosis (hydatid disease) is implemented in practice, consisting in resection of the "protruding dome", resection of the altered and thinned surrounding liver tissue, and omentoplasty using tissue adhesive. The method proposed has definite advantages, namely: readily performed with minimal traumatism, the likelihood of a nonparasitic cyst formation is ruled out, shortened postoperative period and quicker recovery of the patients with resuming of the previous jobs. The indications and contraindications for performing the operation, and possible postoperative complications are discussed. PMID- 7474740 TI - [The Echinococcus and complicated forms of hepatic echinococcosis--their surgical treatment]. AB - A series of 137 patients presenting Echinococcus of which 132 (96.3 per cent) with hepatic location, operated in the clinic of hepatobiliary and pancreatic surgery over the period 1984 to 1994, are analyzed. Right-side location is documented in 59.1 per cent, left-side--in 26.5 per cent, and bilateral--in 14.4 per cent of the patients operated on. In 18.2 per cent it is a matter of a multiple process, and in 27.3 per cent--complicated course. Conservative operative procedures are used in 56.1 per cent, mixed--in 18.9 per cent, and radical--in 25 per cent. Overall mortality in the series amounts to 3.03 per cent; in patients subjected to radical operation--0 per cent, and in cases with serious postoperative complications requiring relaparotomy--similarly 3.03 per cent. Biliary fistulas as postoperative complications are noted in 4.5 per cent, and recurrences in the late postoperative period--in 1.5 per cent. The clinical relevance of the diagnostic methods used, and the indications for performing various operative procedures are discussed. The early and long-term postoperative results are assayed against the background of current literature reports on the issue. PMID- 7474741 TI - [Benign and iatrogenic damages to the bile ducts]. PMID- 7474742 TI - [Our experience with the surgical treatment of cholelithiasis]. AB - The treatment results in 312 patients with cholelithiasis, covering the period 1989 to 1993, are retrospectively analyzed. Overall postoperative lethality amounts to 4.4 per cent. The obtained results are fully comparable in terms of both missed concrements left behind within the extrahepatic bile ducts, and in terms of postoperative complications, associated with the indications for choledochotomy. This is attributed to the therapeutic approach algorithm adopted with respect to the indications for intraoperative cholangiography and choledochotomy. PMID- 7474743 TI - [Chronic recurrent acalculous cholecystitis--apropos 6 clinical cases]. AB - Over the period 1991 through 1994, a total of 120 patients with chronic cholecystitis are operated on. In six of them (5 per cent) no concrements in the gallbladder and biliary ducts are discovered. In 83.3 per cent the echographic study fails to detect concrements in the gallbladder and biliary ducts. All patients are subjected to cholecystectomy, and in all of them markedly expressed adhesions are found. The early and long-term operative results are estimated as very good. The positive therapeutic effect is durable and persists for many years. PMID- 7474744 TI - [The surgical treatment of acute necrotic pancreatitis (the experience of 37 patients)]. AB - The results of the operative management of 37 patients with acute necrotic pancreatitis, covering a ten-year period (1984-1993), are discussed. It is stressed that regardless of the complex conservative and subsequent precisely indicated operative management, acute necrotic pancreatitis remains an exclusively serious and problem disease, accounting for a still very high overall lethality--35.1 per cent according to the case material analyzed. In the authors opinion the most opportune time for surgical intervention is within 5-7 days after the onset of the disease. PMID- 7474745 TI - [The possibilities for organ-preserving operations on the spleen]. AB - Experience had with application of spleen salvaging operations in adult patients is shared. They are used traumatic lesions of the spleen in eleven patients, employing the method of Delany, as modified with the Bulgarian polyamide meshwork Ampoxen. In two of them the hemostasis fails, and splenectomy is resorted to. Preservation of the spleen is accomplished in one patient with chronic pancreatitis where left pancreatectomy with spleen preservation according to the technique of Pradere et al is done. In another female patient, presenting echinococcus of the spleen, echinococcectomy with preservation of the organ is performed. Based on updated knowledge of the role played by the spleen in promoting protection against infection, the strive for its salvage whenever possible is fully justified. PMID- 7474746 TI - [Pancreatoduodenal resection for carcinoid of Vater's papilla]. AB - Papilla Vateri carcinoid is among the very rarely occurring locations of the lesion. The small lumen of the papilla is the underlying cause of becoming clinically manifest with icterus rather early. Accessibility of the process for endoscopic biopsy facilitates the diagnosis. Even histologically it is very difficult to differentiate benign from malignantly altered carcinoids. Only the existence of metastases confirm with certainty their malignant nature. An observation on papilla Vateri carcinoid in a 60-year-old woman, proved preoperatively through cytological study, is reported. Intraoperative--no metastases are discovered. Sparing pancreatoduodenal resection is performed with a good functional outcome, recorded at the checkup examination after two years. PMID- 7474747 TI - [A rare case of gallbladder interposition in the extrahepatic bile ducts]. AB - This is a report on an exclusively rarely met with anomaly, consisting in interposition of the gallbladder within the extrahepatic bile ducts where ducti hepatici open into the gallbladder. Ductus cysticus assumes the role of ductus choledochus, and terminates in papilla duodeni major. Ductus hepaticus communis is completely lacking. The aforementioned anomaly of the biliary system in the female patient reported on is supplemented by a vascular variation, consisting in the following: a. hepatica dex. passes over the isthmus of the gallbladder, and after forming a kink it is directed to hilus hepatis. At the apex of the kink a. cystica divides with blood supply being secured by the latter and the branches of a. hepatica dex. A similar anomaly is described by Nell Krug (2), whereas J. Denes (1) reports on a case where only ductus hepaticus dex. opens into the gallbladder. PMID- 7474748 TI - [Complications of cholelithiasis: a case report]. PMID- 7474749 TI - [Liver resections. Malignant diseases of the liver]. AB - Proceeding from current literature data, personal experience had with resection surgery of the liver, carried out in a series of 64 patients, operated in the clinic of hepatobiliary and pancreatic surgery over the period 1984 through 1994, is analyzed. Thirty-four patients are operated on for benign, and 30--for malignant conditions, with five of the latter group presenting primary tumor of the liver. Seven partial resections, 33 segmentectomies and 24 lobectomies--7 typical and 17 atypical; 6 right and 18 left resections, are performed. Operative lethality in the total series amounts to 0 per cent. The early postoperative lethality in benign diseases is 0 per cent, and in complicated cases--2.9 per cent. In patients with malignant lesions--6.7 and 10 per cent, respectively. The indications for operative treatment, and the possibilities of other therapeutic approaches are discussed. PMID- 7474750 TI - [The strategy and procedure in the surgical treatment of thoracic aortic aneurysms]. AB - This is a summed up report on experience had with patients operated for aneurysms of the thoracic aorta since 1994. Between March 1994 and March 1994, a total of 192 patients undergo surgical treatment for aneurysms of the thoracic aorta (mean age 51.2 years). Of them 152 are men (79.1 per cent). Of the total number 109 cases have acute dissecting aneurysms, and 31--chronic aneurysms. In 114 cases the aneurysms is limited to the ascending aorta only, and in 22-in the descending aorta. The transverse aortic arch is involved in 56 patients. The prevailing preoperative clinical symptoms include: chest pain (171 cases), aortic valve incompetence (108), peripheral ischemia (79) and congestive heart failure (63). Hypertension (148 cases) is the commonest underlying cause encountered. Prosthetic replacement of the ascending or descending aorta alone is done in 32 and 22 patients, respectively. Separate graft replacement of ascending aorta and aortic valve are performed in 54 patients. Bentall-De Bono's operation is used in 43 cases. Partial arch replacement is done in 25 patients, and total arch replacement--in sixteen. The early postoperative mortality in patients presenting acute dissecting aneurysms of the aorta is significantly higher (24.77 per cent), as compared to that in patients with chronic aneurysms (6.02 per cent). There is no significant difference in early postoperative lethality, attributable to the location of the process. PMID- 7474751 TI - [Surgical treatment in patients with diabetic macroangiopathy]. AB - Diabetes mellitus is a disease distributed on a worldwide scale. Although the therapeutic approach to diabetes mellitus and its complications in current use brings about considerable prolongation of the life expectancy in diabetics, the frequency of cardiovascular complications is constantly augmenting; the latter are taken to be the underlying cause of death in this contingent of patients. The lower extremities are usually affected by chronic arterial insufficiency. The aim of this study is to analyze the results of surgical treatment of the occlusive arterial lesions in the lower extremities of diabetics, and specify the indications and type of intervention. Over a 6-year period (1987-1992), 760 arterial operations of the lower extremities are performed. Ninety-six reconstructions are done in diabetics-12.15 per cent. The youngest patient is 39 years of age, and the eldest--80 years. 75 per cent of the patients are given treatment with oral antidiabetic drugs. Most of them have concomitant diseases- 72 cases (75 per cent). Seventy-one (73.96 per cent) present clinical manifestation of rest pain and/or ulcerations, and only 25 cases (26.4 per cent)- intermittent claudication according to R. Fontain. Those with arterial lesions in both segments prevailed--aorto- or iliofemoral lesion. Doppler sonography is performed in all patients with the exception of two studied by angiography. The operative procedures performed include: femoropopliteal--35, aorto-bifemoral--33, aorto- or ilio-femoral bypass-9, arterial reconstructions in both segments--14. The postoperative results are good and fair in 85 cases (88.5 per cent).(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7474752 TI - [The postoperative complications and risk factors in heart operations with extracorporeal circulation]. AB - The analysis covers 150 patients subjected to open-heart surgery in the "St. Catherine" Hospital during the period January through April 1994. The postoperative complications are evaluated with a special emphasis on the risk factors involved. The timing of operation, cardiopulmonary bypass duration, and other predisposing factors are considered. In 105 patients (70 per cent) no complications are recorded in the early postoperative period (12-day postoperative hospitalization, and 30 days after discharge). In the remainder 45 patients (30 per cent), bacterial and abacterial complications develop in 23 cases (15.3 per cent) and 26 cases (17.3 per cent), respectively. Combined bacterial and abacterial complications are observed in 4 cases (2.7 per cent). The outcome of operation is significantly influenced by the timing of surgical intervention, duration of operation and cardiopulmonary bypass, intubation an chest re-exploration. The rest of the factors considered have no essential practical bearing on postoperative complications. In 17.7 per cent it is a matter of noninfectious complications where antibiotic treatment is unnecessary. Subfebrility as a single symptom after the sixth postoperative day is noted in many patients free of complications, and should be by no means considered as an indication for antibiotic therapy. PMID- 7474753 TI - [The results of resection treatment in infrarenal aneurysms of the abdominal aorta over the last 5 years (1989-1993)]. AB - Abdominal aortic aneurysms (AAA) are a major problem in vascular surgery owing to their ever increasing incidence, asymptomatic course and disastrous complications. The positive postoperative results of elective treatment account for a lowered mortality rate, with life expectancy prolonged as much as twice. However, the results of the operative treatment of AAA on an emergency basis are still unsatisfactory. It is the purpose of this paper to describe hitherto experience and success of the clinic with AAA treatment, and inform the medical circles about the increased potentialities of the management of this unexplored, difficult but still curable disease. Seventy-three patients presenting AAA, operated on during the period 1989-1993, are analyzed. The clinical signs and associated diseases, frequency of clinical forms, preoperative factors influencing the risks of forthcoming operations and the long-term results are comprehensively discussed. An asymptomatic clinical form is observed in 60.5 per cent of the patients (Pt = 0.05). Operative lethality is significantly increased in the event of rupture of the aneurysm, its diameter, angina pectoris symptoms and hypertension (P = 0.01). A standard operative technique, described by Creech, is used. Haemonetics Cell Saver is also employed. Thus, the amount of donor blood is reduced by one third of the volume required without any accidents or fatalities. The causes of postoperative death include: ischemic colitis, myocardial infarction and bronchopneumonia. Thanks to the experience and skill of the surgical and anesthesiological teams, the last five years are marked by a low rate of mortality - 3.7 percent for the elective group, and 12.5 percent for the emergency cases.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7474754 TI - [The surgical treatment of vasorenal hypertension]. AB - Reconstructive procedures of renal arteries are routinely advocated for patients presenting documented renovascular hypertension (RVH). It is the purpose of this study to analyze experience had to date, and the results of operative reconstruction of 52 arteries in forty-seven patients, covering a 10-year period. Twenty-seven patients are operated on for RVH. In fourteen patients simultaneous procedures are performed for occlusive disease of the abdominal aorta, three patients are operated simultaneously--ACB, and two--carotid arteries. Aorto-renal bypass is the procedure of choice. In 11 patients TEA is performed for renal artery stenosis. In 45 patients (95.7 per cent) there is a significant improvement of hypertension, and in none of them is renal function lost postoperatively. Hospitalization is prolonged because of postoperative bleeding- two cases, and diffuse peritonitis--one patient, dying in the early postoperative period. Experience with the operative management of RVH shows that operative reconstruction of renal arteries is a method of choice in selected patients, and its indications may be extended to patients undergoing vascular reconstruction of the abdominal aorta. PMID- 7474755 TI - [A comparative assessment of the results of end-to-end and end-to-side anastomoses in arterial reconstructions of the aorto-iliac segment]. AB - The aim of this study to evaluate comparatively the early and long-term outcome of end-to-end (E-E) vs end-to-side (E-S) anastomoses in arterial reconstructions for aorto-iliac occlusive disease (AIOD). The early and late postoperative results of 150 arterial reconstructions for AIOD are evaluated, covering the period 1990 through 1993. All patients present AIOD II-IV clinical stage, randomized in three groups according to type of arterial reconstruction/anastomosis. Group A consists of 60 patients (pts) undergoing aortobifemoral/biprofunda bypass grafting (Ao-2F/P), with all proximal and distal anastomoses performed in the classical E-S fashion. Group B (40 pts)--Ao-2F/P replacement grafting (RG), with all proximal and distal anastomoses performed E E, and simultaneous reconstruction of at least one hypogastric artery and/or inferior mesenteric artery (AMI). Group C (50 pts)--Ao2F/P or unilateral aorto femoral/profunda bypass grafting (Ao-F/P) with EO-S proximal and E-E all distal anastomoses. The aforementioned groups are then compared to 30 pts (group O) free of vascular diseases. All patients are subjected to 18 months average follow-up study (range 6 to 32 months). The postoperative effect, including evaluation of the complete and regional hemodynamics by clinical and Doppler ultrasonography (DS) methods, is assayed. In some cases control angiography is performed as well. The complication rate--restenoses, rethromboses, false aneurysms and amputations- is estimated in percents. Statistically significant higher restenosis and rethrombosis rate is found in group A pts (56 per cent), compared to B (20 per cent) and C (25 per cent) which correlates with the early postoperative results, referred to in the authors' previous study.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7474756 TI - [The 11th anniversary of the founding of the Sveta Ekaterina State University Hospital]. PMID- 7474757 TI - [One-stage operations in pathology of the heart valves and coronary vessels and other general surgical diseases in 124 patients]. AB - Over the last ten years (1984-1994), 124 patients undergo one-stage operations for diseases of the heart valves and coronary arteries and other surgical conditions. This makes 3.38 per cent of the total of 3661 patients subjected to open-heart surgery in the observation period. The indications for undertaking surgical treatment in the series of 124 patients reviewed are classified in three groups, as follows: Group A. Life-threatening conditions due to concomitant surgical disease (bleeding from the gastrointestinal tract, acute calculus cholecystitis, obstructive jaundice, peritonitis, end-stage pregnancy)--16 patients. Group B: Serious non-heart surgical diseases (malignancy, hypersplenism, aneurysm of the abdominal aorta)--47 patients. Group C. Non-heart diseases giving rise to serious complaints and life style deterioration (advanced inguinal hernia, hiatal hernia with gastroesophageal reflux, duodenal ulcus, thyroidism, etc.)--61 patients. The early postoperative mortality and complications rates--2.41 per cent and 3.22 per cent, respectively--do not differ essentially from those in patients with open-heart surgery alone. One-stage surgical procedures after careful assessment of the indications are recommended. PMID- 7474758 TI - [The surgical correction of the right aortic arch with aberrant left subclavian artery and left ligamentum arteriosum--a rare case from clinical practice]. AB - Reports on the surgical management of the vascular ring formed by the right aortic arch with aberrant retroesophageal left subclavian artery and left ligamentum arteriosum are relatively few. The authors present a case of a thirty three-year old woman with cough, dysphonia and stridor without dysphagia. Surgical management which was performed through a bilateral thoracotomy with transversal sternotomy, included division of the ligamentum arteriosum and of the aberrant left subclavian artery and suture of the latter. There was no need to resect the Kommerell's diverticulum, because of lack of any dysphagia despite of the well documented severe compression of the esophagus. The authors experience proved that in difficult cases the bilateral thoracotomy and transversal sternotomy is an useful approach to perform complete surgical repair of this abnormality. PMID- 7474759 TI - [The costoclavicular neurovascular compression syndrome]. AB - This is a description of a great variety of abnormal compressions of nerve, arterial, and rather seldom venous structures at the base of the neck or thoracic outlet, such as supernumerary cervical rib. abnormal ligaments, musculus scalenus anterior hypertrophy and positional alterations, interfering with the normal inter-reactions between first rib and structures overlying it. With a special reference to the hypothetical mechanism of the symptomatology a wide range of denominations of this particular syndrome have been adopted, namely: scalenus anticus syndrome, cervical rib syndrome, costoclavicular syndrome, hyperabductions syndrome and thoracic outlet syndrome--a term gaining widest popularity in the Anglo-Saxon literature. It is the purpose of this study to present experience hitherto accumulated worldwide with the diagnosis and treatment of such pathology, and analyze the authors' experience based on the results of 13 operative interventions for decompressions of the costoclavicular groove against the background neurovascular costoclavicular syndrome. Special attention is focused on the surgical management of this particular pathology using the modern transaxillary approach for resection of supernumerary first rib, thoracic extrapleural sympathectomy--a method successfully introduced in Bulgaria by the authors' team for the first time in 1984. The early and long-term results in 13 patients (7 of them women) operated through transaxillary access over a 10 year period, the complication ensuing and their treatment, as well as comparative assessment with other operative procedures described in the literature, are discussed. PMID- 7474760 TI - [The summary results of percutaneous transluminal coronary angioplasty at the Sveta Ekaterina State University Hospital in 1991-1994]. AB - OBJECTIVES: The article reviews the results of PTCA in 164 patients in Hospital "St Ekaterina" in the period January 1994-April 1994. METHODS: Patients were divided in four groups: I Stable angina II Unstable angina a) primary stabilized b) refractory to medical treatment III Acute myocardial infarction (AMI) IV Silent ischaemia Patients were followed for: 1.) Acute complications a) mortality b) myocardial infarction (MI) c) emergency CABG 2.) Late complications a) recurrent angina b) MI c) elective CABG d) re-PTCA e) late mortality Follow-up period was 6-12 months RESULTS: I group 1) acute complications--early mortality- 0%; acute MI--0%; emergency CABG--5%; 2) Late complications--recurrent angina- 8.4%; MI--1.5%; late mortality--0%; elective CABG--0%; re-PTCA--6.5%. I group 1) acute complications--early mortality--0%; acute MI--6%; emergency CABG--3%; 2) Late complications--late mortality--6%; recurrent angina--10%; MI--6%; elective CABG--1%; re-PTCA--14%. III group 1) acute complications-early mortality--0%; acute MI--17%; emergency CABG--17%; 2) Late complications--recurrent angina--17%; MI--17%; late mortality--17%; elective CABG--0%; re-PTCA--0%. IV group 1) acute complications--early mortality--0%; acute MI--0%; emergency CABG--0%; 2) recurrent angina--18%; late MI--0%; late mortality--0%; elective CABG--0%; re PTCA--18%. CONCLUSIONS: PTCA is a safe and reliable method for revascularization not only in patients with stable angina, but also in patients with acute coronary syndromes. PMID- 7474761 TI - [Intraoperative transesophageal echocardiography in patients with surgical valve repair]. AB - For a period of 13 months (from April 1993 till May 1994) 14 patients (5 men and 9 women) have been examined intraoperatively with transesophageal echocardiography. Nine of the patients were with mitral valve lesion, three of them with mitral valve lesion, complicated with tricuspidalisation, one patient with mitral-aortic valve lesion and high grade tricuspid regurgitation and one patient with aortic valve disease. The purpose of the intraoperative transesophageal echocardiography was to surgical valve repair by measuring the residual stenosis and regurgitation. Intraoperative transesophageal echocardiography evaluation was made by biplane two-dimensional echocardiography, colour Doppler mapping and pulse wave Doppler after CPB before sternum closure. In 13 of the examined patients the effect of the surgical valve repair was assessed by transesophageal echocardiography as very good. In one of the cases intraoperative transesophageal echocardiography indicated aortic and mitral valve replacement because of high grade aortic and mitral replacement, persisting after surgical valve repair of both valves. Intraoperative transesophageal echocardiographic findings were compared with postoperative transesophageal echocardiography before dehospitalisation of the patients. The results of both echocardiographic examinations, correlate to a great extent. The existing difference in assessment of the degree of valvular lesions is due to the specific intraoperative haemodynamic situation. Intraoperative transesophageal echocardiography is a valuable method in assessment of surgical valve repair. PMID- 7474762 TI - [The anatomical characteristics of the a. mammaria interna and their importance for the surgical results in aortocoronary bypass (a report on 2 clinical cases with large intercostal branches of the a. mammaria interna)]. AB - OBJECTIVES: This study attempted to determine the possibility for steal phenomenon after coronary surgery using internal mammary artery (IMA) as graft with unligated big side branches. The internal mammary artery (IMA) is considered to be the conduct of choice in coronary artery bypass grafting (CABH). The more IMA grafts are used, the more increases the significance of IMA anomalies and notably the large side branches for the surgical results. The importance of coronary subclavian steal syndrome is well known. We represent two clinical cases with big intercostal branches visualised at the postoperative coronary angiography, performed to evaluate the coronary and graft status cause of recurrent postoperative anginal pain. After cardiac catheterization we performed ECG stress test as follows: 1. A conventional stress test on treadmill (for control) and 2. Modified stress test loading the chest wall muscles in spine position. We expected to provoke and to evaluate steal phenomenon by the main IMA graft to the big lateral branches. At both of the patients on the arteriographic study IMA graft was patent and both had unusual large (> 50% of the IMA lumen) side branches, reducing the blood flow in the IMA graft. The modified stress test loading the chest wall muscles didn't provoke anginal pain at both patients. At the first (50 years old white male, CABG x 3-July 1193) it was evident non significant ST depression (< 2 mm) in V4-V% (suspicion of silent ischemia). At the second (70 years old white male, anterior MI-1990, GABG x 4-1992) the stress test was interrupted because of fatigue at the arm. No evident significant ecg changes. Despite we couldn't prove steal phenomenon through the angiographic study and the modified ECG stress test we suggest several logic. PMID- 7474763 TI - [An analysis of our experience using the bilateral arteria thoracica interna (ATI) in patients undergoing aortocoronary bypass]. AB - The well established long-term patency of the internal thoracic artery (ITA) makes it a conduit of choice for coronary revascularization. Recent data suggest that patients with bilateral ITA grafts have a greater survival benefit and improved life-style quality, but it is worth noting that increased use of both ITA may have an adverse effect on morbidity and mortality. It is the purpose of this report to assay the first series of patients with bilateral ITAs, and analyze morbidity and mortality rates. In the period April 1992 through January 1994, ten patients undergo coronary revascularization using both ITAs, supplemented by saphenous grafts. In the series analyzed hospital mortality amounts to zero, and there is no perioperative myocardial infarction--neither respiratory deficiency, nor reoperations for bleeding. There is a single case of sternal infection--10 per cent. As shown by the early results, the use of bilateral ITA for coronary artery bypass grafting is a safe procedure, and does not increase operative mortality and morbidity. PMID- 7474764 TI - [The surgical reconstruction of the chest wall using muscle flaps in patients with mediastinitis following heart operations]. AB - An assessment is made of the results of plastic closure of the chest in cases presenting mediastinitis. The analysis covers nineteen patients with a history of mediastinitis following open-heart surgery, treated by the same method of plastic reconstruction of the chest. The technique of plastic closure of the chest using a rotation muscle flap (pectoralis and right rectus abdominis muscles) is described. PMID- 7474765 TI - [Hemophilia B and aortocoronary bypass]. AB - We describe a case of a patient with moderate hemophilia B, undergoing CABG. Substitution therapy was realized with factor IX concentrate (Bebulin-TIM4 IMMUNO). We discuss which of the following factors: hypothermia, hemodilution of heparinisation has a substantial role in alteration of FIX coagulant activity during cardiopulmonary bypass. PMID- 7474766 TI - [The dynamics of the more important indices of the external respiration following heart surgery operations]. AB - A retrospective study comprising patients undergone cardiac surgery is done. In the study some measurable parameters of external breathing are followed up. The measurements are carried out by portable respiratory monitor. The following parameters are included in the study: FVC, FEV1, PEF and FEF 25-75%. The lowering values of these parameters is taken into account and the degree and way to restore these values is discussed. PMID- 7474767 TI - [A dynamic study of plasma fibronectin in patients undergoing heart operations under extracorporeal circulation]. AB - In a preliminary analysis the level of plasma fibronective as well as the total protein and albumin were followed dup in cardiac patients. The dynamic changes of values of the parameters perioperatively and up to the 96-th postoperative hour were taken into consideration. The authors make the conclusion that the fibronectine reduction is not specific and reflects not so much the destruction of the heart muscle but the reparation processes following the surgical trauma. PMID- 7474768 TI - [Anesthesiological and resuscitation problems in the surgical treatment of acute massive pulmonary thromboembolisms]. AB - Eleven patients operated on under extracorporal circulation (ECC) following acutely occurring massive pulmonary emboli were studied. The values of the following parameters were registered: pulmonary artery pressure, pulmonary capillary wedge pressure, intrapulmonary shunt and pulmonary vascular resistance. The measurements were carried out before the surgical procedure and in the postoperative period. The authors defend the opinion that the accurate and timely diagnostics and the correct perioperative management has crucial importance for the determination of the moment for surgical treatment and its successful outcome. PMID- 7474769 TI - [Rupture of the sinus of Valsalva--the anesthesiological and resuscitation problems]. AB - The rupture of the sinus of Valsalva is a rare heart disease with a heavy clinical manifestation, the treatment of which is surgical. It is diagnosed with echocardiography and cardiac catheterisation. The anesthesiologist has to answer many and important questions about anesthetic treatment. There is a little information is the literature on the anesthetic consideration of the patients with a ruptured sinus of Valsalva. On the basis of the results in our hospital the authors want to present the clinical treatment and management of these patients in the perioperative period and to compare these results with other cases like these in world literature. PMID- 7474770 TI - [The use of continuous epidural analgesia in the combined treatment of chronic arterial obstruction in the lower extremities]. AB - Experience had with the application of continuous epidural analgesia (CEDA) in the complex treatment of 33 patients with advanced stage of chronic arterial obstruction in the lower extremities is presented. In fourteen of them (42.4 per cent) CEDA is effected using implantable catheterization system "Periplant", and in the remainder 19 (57.6 per cent)--through nonimplantable "Periflex++". The technique of application of the systems is described. The therapeutic effect is estimated on the basis of: 1) quantitative evaluation of pain perception by a 5 point rating system, 2) acral skin thermometry, and 3) Doppler ultrasound tonometry. Changes in the parameters listed are compared with their counterparts in a control group of 33 patients, and subjected to statistical processing. Proceeding from the obtained results the inference is reached that CEDA is very effective in terms of main relief, and improves perfusion of the ischemic limbs. The implantable catheterization systems contribute to the patient's comfort for rather prolonged period of time, and reduce infection hazards in the vicinity of the spinal cord. PMID- 7474771 TI - [Blood autotransfusion in vascular surgery (predeposited blood)]. AB - The study covers 100 patients, candidates both anamnestically and clinically for autologous blood transfusion. Practically elective patients for reconstructive vascular surgery are concerned. The amount of predonated blood (500 or 1000 ml) depends on the type of operation. A number of indices are followed up: Hb alterations--baseline and after 500 and 1000 ml blood predeposition, Ht alterations--baseline and after 1000 ml blood predeposition, as well as changes in blood protein level following 1000 ml blood predonated. Also, an assessment is made of coagulation factors before and after 1000 ml blood predonation, as well as the optimal longevity of blood storage in refrigerator (the so-called "blood aging"), expressed by K+ levels is stored blood. The studies are carried out in the clinic of vascular surgery of the St. Catherine over the period 1991-1992. As shown by the results autohemotransfusion is a safe, practical and effective procedure, accomplished in one or two stages--only 500 ml blood predonated with 2 3 days interval until operation, and 1000 ml blood predonated in two stages--2-3 days between first and second exfusion, and 3-4 days between the latter and operation. Autohemotransfusion is well tolerated and useful when properly indicated. PMID- 7474772 TI - [Kinetic fundus controlled perimetry with the scanning laser ophthalmoscope]. AB - BACKGROUND: The detection of circumscribed visual field defects due to pathologic macular findings may be difficult even when modern conventional perimetry procedures are used. Using conventional clinical methods there exists no reliable correlation between morphology and function for follow-up and documentation. The Laser Scanning Ophthalmoscope enables a fundus-based examination and therefore allows exact comparison between morphologic appearance and corresponding function. PATIENTS AND METHODS: 50 eyes with pathologic macular findings were examined using a newly developed kinetic perimetry method based on the Scanning Laser Ophthalmoscope. The results were compared to those obtained with other clinical examinations such as Goldmann or Octopus visual field and Amsler-chart. RESULTS: Fundus-based examination with the SLO is preferable for detection especially when small pathologic findings at the posterior pole like macular holes or small serous detachments are combined with reduced stability of fixation. The detection of sharp borders of the scotomas is much better than with conventional perimetry. CONCLUSION: The Scanning Laser Ophthalmoscope allows several different types of fundus-based examinations. Concerning kinetic perimetry we were able to find the borders of scotomas within very short examination times. This method therefore is of valuable help in patients with circumscribed scotomas at the posterior pole combined with deterioration of the stability of fixation and allows documentation for follow-up and further comparison with other clinical findings. PMID- 7474774 TI - [Progress in eyeglass optics]. AB - In this article product developments for ophthalmic lenses are discussed: new materials, designs and coatings. High-index plastic substrates allow to offer corrections which are simultaneously light and thin and for the first time there are high performant plastic photochromic lenses. Head and eye movements with latest generation's progressives are very similar to natural vision behaviour. Special aspheric designs have been developed for comfortable vision for near and intermediate distances. Finally there are new coatings which protect the high quality surfaces of plastic lenses distinctly longer than before. PMID- 7474773 TI - [Congenital familial cornea plana with ptosis, peripheral sclerocornea and conjunctival xerosis]. AB - BACKGROUND: Cornea plana is an extremely rare, congenital hereditary malformation of the corneo-scleral shape. The curvatures of cornea and sclera are nearly equal with an indistinct limbus. In addition to the flatness, there is a peripheral sclerocornea that produces a pseudomicrocornea. The low corneal refraction and the short anterior segment often result in hyperopia. Myopia is also described. Usually the posterior segment is not involved. MATERIALS AND METHODS: A young man of 22 years, his three- and five-year-old sons, and his newborn daughter showed this hereditary abnormality of the cornea. In this uncommon anomaly we measured corneal curvature, refraction, diameter and in three of the four patients echographical length of the bulbi. RESULTS: The family showed an autosomal dominant inheritance of the cornea plana. The corneal refraction was less than 32 diopters. The scleral encroachment caused an oval cornea measuring horizontally between 5 and 6.5 mm, vertically 4 to 5 mm. Additionally a pseudoblepharoptosis and a conjunctival xerosis of the father and his sons was observed, which is not regularly found. A-scan measuring of the bulbi revealed age-related normal values. CONCLUSIONS: There is no evidence for progression of this anomaly during life. No therapeutical consequences are necessary. To preserve a satisfactory function a conscientious orthoptical maintainance should be guaranteed. PMID- 7474775 TI - [Congenital nystagmus in the ophthalmologic automobile driving fitness verdict]. AB - BACKGROUND: A driver with congenital nystagmus (CN) is supposed to have a certain minimum visual acuity and a minimum velocity of reading, with the head directed straight ahead or 10 degrees laterally. The ability to perform different oculomotor tasks depends, however, on individual properties of nystagmus and compensatory mechanisms, as is demonstrated in 4 cases. METHODS: Detailed electrooculographic (EOG) recordings were performed in 4 CN patients, in addition to the examination of visual functions. RESULTS: The visual acuity of patient 1 is sufficient (0.6 on both eyes), yet his retinal image is intermittently destabilized by a periodic alternating nystagmus. Case 2 with high myopic astigmatism and severely reduced visual acuity (right eye: finger counting; left eye: 0.2) compensates for his vertical pendular nystagmus by head nodding and for his manifest-latent horizontal nystagmus by a head turn. The optimal visual acuity of patient 3 (horizontal pendular nystagmus) is 0.3, with an oblique head position and convergence. In other head and gaze positions he has a coarse jerk nystagmus beating in the direction of gaze. Patient 4, with a high myopic astigmatism, microstrabism and manifest-latent fixational nystagmus, has a visual acuity between 0.2 and 0.3 in both eyes. He had been a licensed professional car driver, in spite of contradicting national and international criteria: but his license was refused later. CONCLUSIONS: In patients with CN, the influence of head and gaze position, monocular fixation, convergence and self or object motion on nystagmus intensity and wave forms could be important for driving, because it might influence visual acuity and motion perception. The latter hypothesis still has to be proven in further studies, performed under conditions relevant for traffic. PMID- 7474776 TI - [Keratitis punctata superficialis caused by self-injury]. AB - PATIENT: A 57-year-old patient with diabetes mellitus presented with therapy resistant superficial punctate keratitis in his only eye. Finally self-injury was suspected. After unsuccessful medical treatment of three weeks' duration the disease could be healed promptly after application of an eyepad: the patient admitted self-manipulation. CONCLUSION: In all patients with ocular pathology of uncertain origin, especially in corneal lesions, self-inflicted injury should be part of the diagnostic consideration. PMID- 7474777 TI - [Unexpected clinical course of keratoacanthoma]. PMID- 7474778 TI - [Comment on: "Phototherapeutic keratectomy in recurrence of granular corneal dystrophy after keratoplasty"]. PMID- 7474779 TI - [Clinical experiences with the hand-held keratometer]. AB - BACKGROUND: In this investigation the clinical usefulness of an automatic handkeratometer was compared to well-known methods of corneal power measurement. Moreover, we examined the influence of the patient's position on the results. PATIENTS AND METHODS: One investigator measured 31 eyes of children in the age of 4-11 years (mean 6.3 y.) and 60 eyes of adults in the age of 22-71 years (mean: 48.3 y.) without corneal disorders consecutively by handkeratometer standing and lying, by autokeratometer and computerized videokeratography. Steepest (r1) and flattest (r2) radius, cylinder power and corresponding axis were subjected to statistical analysis. RESULTS: The mean values of the corneal radii (r1/r2) of the children were 7.63/7.82 mm for handkeratometer standing, 7.63/7.84 mm for handkeratometer lying, 7.62/7.84 mm for autokeratometer and 7.70/7.87 mm for videokeratoscope. The cylinder power was 1.11 dpt for handkeratometer standing, 1.09 dpt for handkeratometer lying, 1.23 dpt for autokeratometer and 0.95 dpt for videokeratoscope. Analysis of intraindividual variability produced by the instruments concerning astigmatism (dpt) showed in substraction for comparison of handkeratometer standing with handkeratometer lying 0.02 +/- 0.29 SD, of handkeratometer standing with autokeratometer -0.12 +/- 0.37 and of handkeratometer standing with videokeratoscope 0.16 +/- 0.35. A further comparison concerning cylinder (dpt) of handkeratometer lying with autokeratometer was -0.13 +/- 0.42 and handkeratometer lying with videokeratoscope 0.14 +/- 0.43. Difference comparison of astigmatism measurements in the 95% confidence interval (aspired difference of zero) demonstrated a high statistically significant relationship of handkeratometer standing (p < or = 0.0001) and handkeratometer lying (p < or = 0.0002) with the videokeratoscope. Mean values of the adult eyes (r1/r2) valued 7.56/7.76 mm in handkeratometer standing, 7.56/7.76 mm in handkeratometer lying, 7.57/7.78 mm in autokeratometer and 7.6/7.76 mm in videokeratoscope. The mean cylinder power with the corresponding axis was 1.15 dpt/80.5 degrees in handkeratometer standing, 1.15 dpt/77.8 degrees in handkeratometer lying, 1.15 dpt/78.3 degrees in autokeratometer and 0.95 dpt/75.2 degrees in videokeratoscope. CONCLUSIONS: The results and standard deviations of corneal power measurements of the handkeratometer were independent on patient's position and very similar to those of a common autokeratometer and videokeratoscope. Because of its quick and simple operation the handkeratometer was well accepted for corneal examination of children and can therefore be a useful alternative. PMID- 7474780 TI - [Adhesive eyelid weights ("reversible lidloading") for treatment of lagophthalmos]. AB - BACKGROUND: In facial palsies with lagophthalmic keratopathy an hour-glass dressing is indicated, which has the disadvantage of a moisture chamber with visual impairment by steaming up. After weeks of placement dermal irritations are possible. An alternative is the gravity dependent lidloading, using gold implantation in cases of irreversible lagophthalmos. METHODS AND PATIENTS: Lead weights of 0.8 to 2.0 g are glued to the upper lid with an adhesive layer, that is well tolerated by the skin (Combihesive*), or by a simple foil that is glued on both sides (Tesafix). This results in a lidclosure without impairment of lid opening. After tests on 10 normal persons the described method was placed on 22 patients with lagophthalmos and compared with hour-glass dressing. The new method was applied to 36 additional patients. RESULTS: The dynamic lead weight is effective and well-tolerated. It was more accepted cosmetically especially during the day because of better cosmetic appearance and better visual function than was the hour-glass dressing. These latter dressings are still important for use in serious cases during the night. CONCLUSION: lidloading with lead weights that are glued by an adhesive layer to the upper lid can be recommended as a new method in cases of reversible lagophthalmos or as a preparatory step before gold implantation. PMID- 7474781 TI - [Granuloma annulare of the eyelid]. AB - BACKGROUND: Granuloma annulare is a chronic inflammatory disease of connective tissue, manifested clinically by small, firm nodules. It is observed only rarely around the eyes. PATIENTS AND METHODS: We present four patients aged 33 to 66 years with papulomatous lesions of the eyelids. The lesions were mildly or not symptomatic. Biopsies were taken. There was no association with any systemic inflammatory disease. RESULTS: Histology was characteristic for granuloma annulare in all four cases. The clinical course was benign. CONCLUSION: When faced with papulomatous lesions of the eyelids, the ophthalmologist may consider a diagnosis of granuloma annulare in patients of any age. While granuloma annulare is generally observed in young persons, one of our patients was aged 66 years. PMID- 7474782 TI - [Therapeutic hydrophilic bandage lenses after perforating keratoplasty in severe eye chemical burns]. AB - BACKGROUND: The prognosis of penetrating keratoplasty after severe eye burns is uncertain. Beside of immune reactions the outcome is determined by surface problems. PATIENTS AND METHODS: Between July 1991 and October 1993 in 15 patients (16 eyes) with grade IV eye burns penetrating keratoplasties with large diameters (11-16 mm) were carried out. Cultured corneas were used with an intact epithelium. In 9 eyes hydrophilic bandage lenses (Geaflex 70, Fa. Wohlk, Kiel) were applied initially, in the remainder 7 eyes within 7 days postoperatively. RESULTS: The lens radius best suited was found out by trial, because corneal topography was not possible for many weeks. In 12 (75%) eyes the lens had to be fitted steep, with a radius from < or = 8.7. A change in lens radius during the healing course was rare. The frequent use of artificial tear drops was important, because many eyes showed clinical manifestations of dry eye syndrome. Complications during soft contact lens wearing were rare. In 5 eyes microbiological examination was positive, but successfully treated. Under the protection of soft contact lenses 9 eyes maintained an intact epithelium. In the remainder eyes severe vascularisation with large persistent epithelial defects occurred mostly as a consequence of immune reactions. These keratoplasties developed on growth of inflammatory pannus or progressive ulceration. The average of the follow-up time was 22 months. CONCLUSION: The use of large diameter keratoplasties in combination with hydrophilic bandage lenses proved to be successful to maintain the integrity of the epithelium in these high-risk keratoplasties. The prognosis of these transplants is, however, determined by immune reactions. PMID- 7474783 TI - [Development of pars plana vitrectomy. My personal contribution]. PMID- 7474784 TI - [Intraocular silicone lenses and silicone oil]. AB - BACKGROUND: Foldable silicone intraocular implants are becoming more popular in conjunction with small incision phacoemulsification. PATIENTS: We observed three patients with silicone oil droplets adherent to the posterior surface of the silicone implant, following silicone oil removal after preceding vitreoretinal surgery with the installation of silicone oil. These droplets could not be dislodged intraoperatively either with focal aspiration or irrigation. They interfered with the patients subjective visual acuity. For this reason, we performed an exchange of the silicone intraocular implant against a PMMA one piece lens in all three cases. RESULTS: This procedure resulted in an increase of the visual acuity. The mechanism of adherence between the two silicone polymers is finally not known. CONCLUSIONS: With increased use of implantable silicone intraocular lens and silicone oil this complication may be encountered more frequently. Therefore, implantation of silicone lens in vitreoretinal high risk eyes should avoided. PMID- 7474785 TI - [Endothelial keratopathy in pseudoexfoliation syndrome: quantitative and qualitative morphometry using automated video image analysis]. AB - This prospective study intended to quantify and classify morphological changes of the corneal endothelium in pseudoexfoliation syndrome (PSX) after having tested reproducibility and validity of a new automated technique for analysing corneal endothelium. PATIENTS AND METHODS: We used a contact specular microscope combined with a video camera (Tomey EM-1000) and a computer (IBM compatible PC, 486DX33) with suitable software (Tomey EM-1100, version 0.94). Video images of corneal endothelium (area: 0.312 mm2) are passed directly into the computer input by means of a frame grabber and are automatically processed. Missing or falsely recognized cell borders are corrected using the mouse. We examined 85 eyes with PSX and 33 healthy control eyes. At first, retest-stability and validity of the cell density measurements were assessed in the PSX-eyes. A qualitative analysis of the corneal endothelium followed. RESULTS: The cell density measurements showed a high retest-stability (reliability coefficient r = 0.974). The values of the automated method (2040 +/- 285 cells/mm2) and those of manual cell counting (2041 +/- 275 cells/mm2) did not differ significantly (p = 0.441). The mean difference was 3.1 +/- 2.4%. Comparing the 85 PSX-eyes (2052 +/- 264 cells/mm2) to the 33 control eyes (2372 +/- 276 cells/mm2), there was a significant reduction of cell density (p < 0.001). The cell density of the 69 PSX-eyes with glaucoma (2014 +/- 254 cells/mm2) was significantly lower than that of the 16 PSX eyes without glaucoma (2214 +/- 251 cells/mm2) (p = 0.008). Eighty-five percent of the 85 PSX-eyes showed polymegalism, 77% pleomorphism; 68% had white deposits and 42% guttae. White deposits and guttae were significantly more frequent and more intensive in PSX-eyes than in control eyes. PSX-eyes with and those without glaucoma showed no significant differences concerning the four qualitative parameters. CONCLUSION: The automated method for analysing corneal endothelium quickly provides reproducible and valid results using the correction mode of the software. Semiquantitative analysis of qualitative parameters permits a more differentiated assessment of keratopathy in pseudoexfoliation syndrome than does mere consideration of endothelial cell density. Both evaluations are recommended to assess the risk of a diffuse endothelial decompensation before intraocular surgery. PMID- 7474786 TI - [Induced astigmatism in extracapsular cataract extraction with tunnel incision and various wound closures]. AB - BACKGROUND: For planned extracapsular cataract extraction the no-stitch technique with 11-mm tunnel width has been well established. Four modifications of wound closure were performed to further reduce surgically induced astigmatism. PATIENTS AND METHODS: In this prospective study we controlled 250 eyes of 250 consecutive patients 4 months after surgery with four different wound constructions: sutureless wound closure (n = 70), singular perpendicular suture (n = 100), cross suture (n = 40) in 12 o'clock position or sutureless wound closure in temporal position (n = 40). RESULTS: Surgically 'Induced Astigmatism' was for eyes with preoperative 'With the Rule Astigmatism' (vs Against the Rule astigmatism), operation in 12 o'clock position and sutureless wound closure 2.22 +/- 0.77 D (1.66 +/- 0.94 D), with perpendicular suture 1.66 +/- 0.93 D (1.24 +/- 0.82 D), and with cross suture 1.47 +/- 0.96 D (0.9 +/- 1.13 D). Temporal incision was only performed in preoperative 'Against-the-Rule-Astigmatism' eyes and resulted in 0.6 D of 'Induced Astigmatism'. Preoperative average astigmatism was 0.86 +/- 0.68 D (1.01 +/- 0.95 D). CONCLUSIONS: For preoperative 'With the Rule Astigmatism', operation in 12 o'clock position and singular perpendicular suture and for 'Against the Rule Astigmatism' (especially > 1.5 D) temporal incision is recommended. PMID- 7474787 TI - [2-dimensional mapping and retinal and papillary microcirculation using scanning laser Doppler flowmetry]. AB - PURPOSE: To present clinical applications of a new non-invasive method imaging in a high-definition the topography of perfused retinal vessels. METHOD: By a combination of a laser Doppler flowmeter with a scanning laser system the perfusion of the retina and the optic nerve head is visualized and quantified. The principles of measuring blood flow by Laser Doppler Flowmetry are based on the optical Doppler effect: laser light scattered by a moving particle is shifted in frequency by an amount delta f. Our data acquisition and evaluation system is a modified laser scanning tomograph. The technical data are: retinal area of measurement 2.7 mm x 0.7 mm, 10 degree-field with 256 points x 64 lines, measurement accuracy 10 microns, wavelength 670 nm and 790 nm, light power 100 microW, data acquisition time 2,048 s. Every line is scanned 128 times by a line sampling rate of 4,000 Hz. By performing a discrete Fast Fourier Transformation over 128 intensities of each retinal point the laser Doppler-shift is calculated for each retinal point. With these data a 2-D map with 256 x 64 points of the retinal perfusion is created. The brightness of the picture-point is coded by the value of the Doppler shift. We estimated the reliability and the validity of the method. Perfusion-pictures of the superficial retinal layer and in the optic nerve head were presented. RESULTS: The reliability-coefficients r1 of "Flow", "Volume" and "Velocity" were 0.85, 0.83, and 0.85 respectively. The blood flow measurements by the presented method ("Scanning Laser Doppler Flowmetry") in an artificial capillary gave a linear relationship (r-value 0.973, p < 0.00001) between defined blood velocities and the measured blood flow. By the confocal technique, dependent on the focus, capillaries of the retinal superficial vasculature of the optic nerve head became visible with a high resolution. Off line the blood flow of areas of 110 microns x 110 microns were calculated in terms of laser Doppler flowmetry. CONCLUSION: "Scanning Laser Doppler Flowmetry" facilitates the visualisation of perfused retinal capillaries and vessels in high resolution. The representation of the function of the retinal circulation by SLDF leads to an image similar to the anatomical situation. The 2-dimensional mapping of local blood flow leads to a physiological picture of the retinal perfusion with visible vessels and capillaries. PMID- 7474788 TI - [Biological corneal replacement--an alternative to keratoplasty and keratoprosthesis? A pilot study with heterologous hyaline cartilage in the rabbit model]. AB - BACKGROUND: In patients with corneal opacity caused by diseases like okular pemphigoid, Stevens-Johnson-Syndrome or burns, visual rehabilitation can only be achieved with keratoplasty or keratoprosthesis. The first has generally a poor prognosis in these circumstances, and the latter bears several problems in the postoperative course as well. A biological, corneal replacement could possibly combine the advantages offered by keratoplasty (no interaction between host tissue and plastic) and keratoprosthesis (more permanent transparency). Theoretical considerations and practical experience suggest that the best candidate would be hyaline cartilage. MATERIAL AND METHODS: Bovine sterni were used to obtain circular cartilage slices which were approximately 60 microns thick, having a diameter of approximately 6 mm. These slices were implanted in one eye each of 4 rabbits instead of the natural cornea. The eyes were enucleated after 8 to 29 days. RESULTS: The cartilage remained so transparent that the iris and to a certain extent the fundus blood vessels could be visualized. Connective tissue served to bind the host cornea to the cartilage, but did not infiltrate the cartilage. Perforation was observed in 3 animals. The extent of epithelialization of the cartilage slices ranged form ca. 30% to 95%. None of the animals exhibited an intraocular inflammation. The cartilage did not proliferate and remained avascular. Secondary intraocular changes, with the exception of anterior synechiae, could not be detected. CONCLUSIONS: A tissue for biological corneal replacement has to fulfill many requirements which to a certain extent can be achieved with the use of hyaline cartilage. Despite numerous, unsolved problems, a biological corneal replacement as an alternative to keratoplasty and keratoprosthesis in cases of corneal blindness does not seem out of imagination. PMID- 7474789 TI - [Lens astigmatism in connection with implantation of a foldable one-piece silicone lens in a patient]. AB - BACKGROUND: The use of one-piece plate-haptic Silicon lenses has become more frequent especially in combination with small incision cataract surgery. Different complications as before are to be expected. CASE REPORT: Six weeks after cataract surgery and implantation of a one-piece plate-haptic Silicon lens a lens astigmatism of 3.5 dpt was noted. The astigmatism occurred after successfully performed retinal detachment surgery in this eye. The development of the astigmatism was considered to be due to a capsular shrinkage of the anterior capsular ring. The retinal detachment surgery was performed with the use of SF6 gas, which is known as a factor for increasing inflammation. After a Nd: YAG incision of the anterior capsule the astigmatism disappeared immediately. CONCLUSION: Capsular shrinkage can induce a significant lens astigmatism in combination with soft one-piece plate-haptic Silicon lenses. Intraocular inflammation seems to play an important role. Astigmatism after implantation of one-piece plate-haptic Silicon lenses in combination with signs of capsular shrinkage can be explained by lens astigmatism only. PMID- 7474790 TI - [Simulation of anterior ischemic optic neuropathy by optic nerve sheath meningioma]. AB - BACKGROUND: The optic nerve sheath meningioma usually presents with a diffuse atrophy of the disc and a diffuse depression of the visual field. Applying this rule too rigidly can lead to a false diagnosis. PATIENT: A 45y-old-woman presented with atrophy of the upper disc and a corresponding altitudinal field defect in her right eye. AION was diagnosed. Seventeen years later the patient returned with an exophthalmus. The disc was atrophic and a shunt vessel connected the central retinal vein with the choroid. CT revealed an orbital mass. The diagnosis of an optic nerve sheath meningioma was made and confirmed at surgery. DISCUSSION: The meningioma may have occluded an upper posterior ciliary artery, mimiking an AION. Alternatively, the meningioma may have destroyed the fibers running through the upper half of the disc and caused swelling of the remaining fibers in the lower half of the disc. CONCLUSION: In the combination of swelling and atrophy of one disc an optic nerve sheath meningioma should be considered, even if the two components are located in different sectors. In optic nerve sheath meningioma vision decreases insidiously as opposed to the acute drop typical for anterior ischemic optic atrophy. PMID- 7474792 TI - [Varicose vein of the iris]. AB - A 56-year-old man presented with a pigmented iris nodule which showed prominent vessels and enlargement within 6 weeks. The tumour caused recurrent hyphemata. Differential diagnosis included a pigmented iris hemangioma and a malignant melanoma. A sectoriridectomy was performed. Histology revealed a stromal cavity filled with fibrin and abundant erythrocytes. A diagnosis of an iris varix was established. However, it has to be pointed out that the distinction between a varix of the iris and an iris hemangioma may be difficult not only clinically but also histologically, and that iris melanomas may produce hemangioma-like patterns. PMID- 7474791 TI - [Evaluating vision after photorefractive keratectomy within the scope of legal police service requirement 300]. AB - BACKGROUND: The Polizeidienstvorschrift (PDV) 300 is a regulation which prescribes visual requirements for applicants to the German Police Service. Among other demands, uncorrected visual acuity (VA) of both eyes must be at least 0.5 before age 18 and at least 0.3 after age 21. The spherical equivalent of spectacle correction is ordained to fall into the range between -2.0 and +3.0 dpt. To date, recommendations for the assessment of excimer laser photorefractive keratectomy (PRK) with a view to Police Service applicants are still lacking. PATIENTS: Two applicants with myopic astigmatism were treated by PRK in order to meet the refractive requirements for entering the German Police Service. The first patient was judged fit for service two weeks after PRK was performed on his right eye. At this time uncorrected VA was 1.0 VA under glare conditions and contrast sensitivity had not been tested. The second patient had a stable postoperative refraction with an uncorrected VA of 1.0 in both eyes at six months postoperatively. VA under glare conditions and contrast sensitivity were normal. However, the medical service of the police authority demanded an additional follow-up of six months before judging this applicant fit for service. CONCLUSIONS: Before excimer laser PRK is performed in myopic police service applicants, a careful examination should rule out disorders of the visual system which are not related to the refractive error and may preclude admission into the police service. After PRK in this group of persons, a postoperative standard follow-up of at least six months should be demanded. Since fitness for night driving, self-defence ability and good orientation under difficult visual conditions are mandatory for police officers, any ophthalmologic certificate should make definite statements about glare and contrast sensitivity. PMID- 7474793 TI - [Color doppler ultrasonography in diagnosis of ocular and orbital tumor in children]. AB - PURPOSE: To evaluate the usefulness of USG-CD in the assessment of ocular and orbital tumors in children. METHODS: 24 children with focal changes in eyeballs and orbita were examined. The results of USG-CD were compared with those of computer tomography and angiography. Diagnosis was confirmed during surgery or by biopsy. RESULTS: Typical USG-CD pictures of haemangioma, melanoma, retinoblastoma and also pseudotumors caused by haematoma and inflammation were described. The sonographic examination enabled to determine the placement and extent of the lesions, the Doppler option allowed to evaluate their blood-supply and in some cases, e.g. in haemangioma, to state a diagnosis. PMID- 7474795 TI - [The role of Chlamydia trachomatis in the etiology of conjunctivitis]. AB - PURPOSE: To turn attention to the etiological importance of chlamydia in chronic conjunctivitis. METHODS: From December 1993 to November 1994 the authors examined 95 patients with chronic conjunctivitis for the presence of Chlamydia trachomatis antigens in conjunctival epithelial cells using direct immunofluorescent antibody tests. RESULTS: The test was positive in 23 patients and most of them responded positively to general tetracycline treatment combined with locally used Sulfacetamide or Floxal. CONCLUSION: Chlamydia trachomatis are common causes of conjunctivitis in adults. PMID- 7474794 TI - [Surgical methods in patients with cataract and myopia]. AB - PURPOSE: To present own experiences in cataract surgery in myopic eyes. METHODS: 72 eyes of patients, aged 41-84, with 4-year follow-up were analysed. According to the degree of myopia, the patients were divided into 3 groups: MP up to -4.0 D (29 eyes), mp from -4.25 D to -8.0 D (27 eyes) and MP > -8.0 D (16 eyes). Extracapsular cataract extraction was performed in 55 eyes (76.4%) and intracapsular in 11 (15.2%). Posterior IOLS were implanted in 30 eyes (41.6%), anterior IOLS (including 4 secondary ones) in 26 (36.1%) and 16 eyes (22.2%) were operated on without IOLS implantation. RESULTS: Good visual acuity (> 0.5) was achieved in 73.6% of cases, useful visual acuity (> 0.1) in 89%. The results as well as the postoperative complications were compared with the control group of non-myopic eyes. Glaucoma was observed in 18% of myopic eyes and in 10% of non myopic but it was not significant. CONCLUSION: The safest method of cataract surgery in myopic eyes is extracapsular extraction with posterior IOL implantation. These eyes should be operated on by very experienced surgeons. PMID- 7474796 TI - [Incidence of hypersensitivity to antibiotics and preservatives in eye drops]. AB - PURPOSE: To elaborate a set of contact allergens present in eye drops which could precipitate allergic conjunctivitis. Such set has not been yet described in literature. METHODS: Basing on the most frequently used preparations, a set of 15 different substances: drugs and preservatives, was prepared to test the patients. RESULTS: Among the 97 persons who underwent the testing, positive results were observed in 35.5%. The hypersensitivity was found for 11 of the allergens, the most often for neomycin, gentamycin, merthiolate, benzalkonium chloride, and then consecutively for kanamycin, sulfathiazole, oxytetracycline, chloramphenicol, penicillin, boric acid and parabens. PMID- 7474797 TI - [Results of strabismus surgery in adolescents and adults: cosmetic or functional recovery?]. AB - PURPOSE: To answer the question, whether it is possible to achieve a functional improvement after strabismus surgery in adults. METHODS: The study comprised 71 patients, aged 13-53, in whom after strabismus surgery, deviation was reduced to +/- 4 degrees. The degree of binocular vision was examined with weak and strong dissociating tests. RESULTS: Complete binocular vision was achieved in 25% of cases, partial with peripheral fusion in 51%. The degree of binocular vision depended on the following factors: type of squint and time of its development, degree of amblyopia, refraction and presence of binocular vision before the surgery. In some cases binocular vision occurred immediately after surgery, in others in a long follow-up. Stabilization of surgical effect was observed in the cases with peripheral fusion. CONCLUSION: Strabismus surgery in adults could be a chance for functional recovery and not only for aesthetic improvement. PMID- 7474798 TI - [Advantages and disadvantages of periocular analgesia]. AB - PURPOSE: To evaluate the advantages of local analgesia in the form of posterior periocular blockade in comparison with retrobulbar blockade, used till now. METHODS: During last 5 months 277 operations were performed in local analgesia, including 225 periocular and 52 retrobulbar. RESULTS: Periocular analgesia is sufficient in cataract and antiglaucomatous surgery. The following complications were observed: palpebral haematoma, subconjunctival infiltration and transient visual loss. CONCLUSION: In comparison with retrobulbar periocular technique of analgesia is less painful, easier to perform and safer in terms of perforation of the eyeball. Yet its inconvenience is that it must be done about 20 minutes before the surgery, connected with the delay effect of Bupivacaine. PMID- 7474800 TI - [Twenty year observation of a severe case of bilateral retinoblastoma]. AB - PURPOSE: To present a case of a girl with bilateral retinoblastoma, who, despite of the continuing development of the disease, survived for twenty years. METHODS: Neoplasmatic lesions, diagnosed in the child's left eye at age of 2, were so advanced that it was necessary to enucleate the eyeball. Relatively late, i.e. after 7 years, the tumour appeared in the second eye. This eye was treated with xenon-arc photocoagulation and 60Co irradiation. RESULTS: In spite of many years of treatment the lesions were progressing. Enucleation of the second eye was much delayed due to the lack of consent of the parents. 15 months after enucleation a recurrence appeared in the eye socket. Surgical treatment was supplemented by irradiation of the eye socket and by chemotherapy. After 20 years of intensive treatment the patient died because of generalized neoplasmatic disease. CONCLUSION: This case confirms the necessity of prolonged, careful observation of patients with retinoblastoma. PMID- 7474801 TI - [Hamartoma of the optic disc in a 17-year old boy]. AB - BACKGROUND: Hamartoma of the optic disc is a rare disease but a very important one for differential diagnosis with choroidal melanoma. METHODS: A case of a boy, 17, referred to the clinic with suspicion of intrabulbar tumor is presented. Basing on the clinical picture ultrasonography and fluorescein angiography studies, diagnosis of retinal and pigment epithelium hamartoma of the optic disc was established. Laser treatment was applied. RESULTS: During 4-year follow-up visual acuity and ophthalmoscopic picture of the lesion has not deteriorated. PMID- 7474802 TI - [Retinal detachment after accidental intra-vitreous Depo-Medrol injection. A case report]. AB - BACKGROUND: Accidental Depo-Medrol injection into vitreous cavity is a very rare complication and early retinal detachment, associated with it, has a very poor prognosis. METHODS: A case of a women, 34, treated on account of anterior uveitis, in whom during Depo-Medrol injection perforation of the eyeball was done is presented. The drug was injected into the vitreous and retinal detachment developed. The severe changes observed in the fundus of the eye are described. In the treatment vitrectomy, encirclement, radial buckling and endolaser were applied. RESULTS AND CONCLUSION: Retinal attachment with good functional results was achieved, which indicates the importance of early surgical treatment in such cases. Vitrectomy allows to remove the noxious substance from the eye, to see and evaluate the retinal changes and to apply a proper treatment. PMID- 7474799 TI - [Retinitis due to cytomegalovirus in AIDS]. AB - Clinical course of 4 AIDS patients suffering from CMV retinitis is presented. Two patients were treated with ganciclovir, one died before starting the treatment and one was vitrectomized with silicone oil injection. Of the two patients treated with ganciclovir one improved significantly when the other continued to deteriorate and died after 8 months of follow-up. Early diagnosis and treatment of patients with zone I involvement gives the best chances for improvement. Mean life expectancy in aids patients after diagnosis of CMV retinitis is 7-10 months. PMID- 7474803 TI - [Retinal antigens in eye diseases--clinical value of diagnostic tests]. AB - Current review of data concerning the role of antigen S, IRBP and antigen CAR in various ocular diseases, including, uveitis of unknown etiology, birdshot choroid retinopathy, vasculitis, diabetic retinopathy and retinitis pigmentosa. PMID- 7474805 TI - [Results of radial keratotomy as evaluated by patients]. AB - PURPOSE: To present the preliminary report concerning the problem how the patients who underwent radial keratotomy evaluate the surgery itself and its results. METHODS: Questionnaire forms were sent to 60 patients operated on in the years 1990-1993. There were 11 questions about the decision of operation, its course, possibly complications and results. RESULTS: Forms were returned by 49 patients (81.7%). 94% of patients were satisfied with the results achieved. There were great differences between the patients and doctors as to the medical and professional indications for radial keratotomy. According to patients they exist in 73.5% cases, from the medical point of view--only in 45%. PMID- 7474804 TI - [Contribution of immunologic mechanisms in endogenous uveitis]. AB - It has long been known that eye is an immunologically privileged organ. This is concerned with central and peripheral mechanisms that control the eye immunological tolerance. New models of human inflammatory eye disorders were induced in laboratory animals by immunization with photoreceptor antigens (EAU) and retinal pigmented epithelium antigens (EAAU and EAPU). Continual investigations on eye immunological response mechanisms indicated the central role of lymphocyte T-helper, although reactions with circulating immune complexes cannot be excluded. Individual susceptibility to autoimmune uveitic disorders is genetically controlled and is regulated by MHC-antigens and by non-MHC dependent factors, and the latter ones seem to play a more important role. PMID- 7474806 TI - [Optimal ocular correction for computer operators]. AB - Different methods of correction of refractive errors with glasses were discussed in the paper. The problem of the use of antireflection coating and tinting or color coating of the lenses was also presented. PMID- 7474807 TI - [Role of acid-base balance of the blood in the pathogenesis of acute gastroduodenal ulcers in surgery patients]. PMID- 7474808 TI - [Tactics of treatment of patients with arrhythmia]. PMID- 7474809 TI - [Prophylactic use of lidocaine in the acute period of myocardial infarction]. PMID- 7474810 TI - [Disseminated intravascular coagulation syndrome in surgical practice]. PMID- 7474811 TI - [Treatment of respiratory insufficiency in chronic nonspecific lung diseases]. PMID- 7474812 TI - [Current concepts concerning functional dyspepsia syndrome]. PMID- 7474814 TI - [Bone marrow transplantation in beta-thalassemia major]. PMID- 7474813 TI - [Gastric erosion, erosive gastritis: their forms and approaches to treatment]. PMID- 7474815 TI - ["Klinicheskaia Meditsina" is 75-year-old]. PMID- 7474816 TI - [Use of epithalamin in climacteric myocardiopathy]. PMID- 7474817 TI - [Effectiveness of systemic thrombolytic therapy in acute myocardial infarct]. PMID- 7474818 TI - [Use of bonnecor in the treatment of ventricular arrhythmia]. AB - The effectiveness of the new antiarrhythmic drug bonnecor was tried in 59 patients with ventricular arrhythmia. High antiarrhythmic activity of the drug was recorded in the course doses 150 to 250 mg/day. Bonnecor effect is dose dependent: the number of ventricular extrasystoles reduced by 75% in 62 and 74% of the patients receiving 150 mg/day and 20 mg/day, respectively. The highest effect was achieved in high-grade ventricular extrasystole. Side effects were occasional: headache lasting 1-2 hours 40-50 min after bonnecor administration, slow atrioventricular conduction (2 cases), sleep disorders (2 cases). PMID- 7474819 TI - [Effects of smoking and coffee consumption on indicators of arterial pressure in 24-hour monitoring]. AB - 27 essential hypertensives were included in the study to evaluate the influence of cigarette smoking and coffee drinking on 24-hour blood pressure and its variability. The patients were untreated with antihypertensive drugs for at least 4 weeks. All the patients were coffee drinkers (4-5 cups a day) and cigarette smokers (during 25.2 +/- 7.4 years before study and about 10-20 times a day during last year). 24-hour ambulatory blood pressure monitoring (ABPM) was performed 3 times a week at one-day intervals. During the first and third ABPM the amount of coffee and cigarettes as well as schedule of both were equal, during the 2nd ABPM-smoking and coffee were prohibited. There were no significant differences between mean BP values and variability during the 1st and 3d ABPM. Variability of 24-h and daytime systolic blood pressure (SBP) was higher in the 1st--and 3d vs. 2nd ABPM. The 2nd ABPM revealed a significant SBP decrease at daytime (152.5 +/- 9.2 mm Hg vs. 147.2 +/- 8.9 mm Hg, p < 0.05) without changes in nighttime as well as in the whole 24-h period. Values of diastolic BP did not change. The results suggest that cigarette smoking and coffee drinking increase mean daytime and overall 24-hour SBP values and variability as well as heart rate, while diastolic BP is more stable. PMID- 7474820 TI - [Functional evaluation of the condition of patients with peptic ulcer and its changes during treatment]. PMID- 7474821 TI - [Genetic aspects of heart lesions in patients with chronic alcoholism]. AB - Parents and their children from 40 families of the first generation and 38 families of the second generation were examined genetically for catalase activity in the serum. It was found that children of the first generation suffered from chronic alcoholism more frequently in the families in which either mother or both parents abused alcohol. Low activity of serum catalase occurred in those children of the second generation, whose both parents were chronic alcoholics and had low catalase activity, especially of their mother. If the son was a chronic alcoholic, the inheritance of low catalase activity from his mother caused alcoholic damage to the heart. The risk was still higher in the second generation, especially in girls because of X-chromosome effect. PMID- 7474822 TI - [Hyperinsulinemia and severity of cardiovascular disorders in generalized lipodystrophy syndrome]. AB - The examination of 40 patients with generalized lipodystrophy elucidated the dependence of the severity of cardiovascular disorders in these patients on the immunoreactive insulin/C-peptide index. In high values of the latter cardiovascular disorders occur more frequently. The role of insulin in pathogenesis of essential hypertension, chronic IHD is assessed. PMID- 7474824 TI - [Differential diagnosis of exudative pleurisy]. AB - The authors propose signs of high informative value (eosinophilia of the pleural fluid, high amylase concentrations in the pleural exudate) to be used as help in differential diagnosis of pleurisy. Eosinophilic pleurisies are caused by injuries and helminthic invasions, whereas elevated levels of amylase in the pleural fluid are indicative of chronic pancreatitis. PMID- 7474825 TI - [Effects of smoking on motility of the upper gastrointestinal tract in healthy persons and patients with peptic ulcer]. AB - 59 male volunteers (16 healthy and 43 ulcer subjects) entered the study on effects of smoking on gastrointestinal motility. The latter was assessed within 12-50 hours by means of probes fitted with tensiometers. Smokers, especially heavy smokers, regardless of the disease presence, had more regular phase alterations in the cycles of the motility and more regular rhythm of phase III, more frequent involvement of the stomach in frontal motor activity. A direct effect of the cigarette smoked on gastric and intestinal motility was associated with condition of the patient and time after meal. PMID- 7474823 TI - [Chronosensitivity to adelphane-esidrex and sinepres and effectiveness of treatment of patients with hypertension]. AB - 24-h trends in central and peripheral circulation in response to pharmacological tests were registered in 40 patients with stage II essential hypertension (EH). Rhythmic patterns of hemodynamic parameters (heart rate, systolic pressure, stroke volume, etc.) sensitivity to adelphane-esidrex were found. Hypotensive effect of this drug was maximal in morning hours because of the fall in arterial pressure due to reduced peripheral resistance. Sinepres chronotherapy given to 18 EH patients turned out superior to conventional treatment (1 pill 3 times a day) given to 20 EH patients. When taken into consideration, chronosensitivity to the above drugs may bring about a reduction in daily and course doses and earlier occurrence of a hypotensive effect. PMID- 7474826 TI - [Lipid content of the bile in diseases of the biliary tract in male and female Evenkis]. AB - A total of 156 Evens and 162 Evenks were examined in the reindeer-breeding regions of Evenkia and Yakutia in the Extreme North of the country. The Evens suffering from cholecystitis presented with higher saturation of the B and C duodenal bile with cholesterol than patients with dyskinesias. Such regularities were not observed in the Evenks. These findings permit a conclusion that the ethnic and geographic factors influence the lipid composition of the bile in indigenous population of the Extreme North in case of diseases of the biliferous ducts. PMID- 7474827 TI - [Therapeutic tactics and results of prostatic adenomectomy in the presence of aggravating factors]. AB - Outcomes of radical surgery for prostatic adenoma have been analyzed for 426 patients divided into 3 groups. Group 1 included 160 patients subjected to urgent adenomectomy for acute urine retention, group 2 comprised 166 patients with severe intercurrent diseases subjected to one- and two-stage radical surgery, group 3 of 100 patients was free of intercurrent diseases. Urgent and delayed (within 72 hours and 5-10 days, respectively) radical adenomectomies are thought valid in management of acute urinary retention in patients with prostatic adenoma. Adequate preoperative preparation in deficient circulation, diabetes mellitus and other preexisting or concurrent diseases increased by 20-30% indications to the operation. Good outcomes were reported in 90%, 81.4% and 94% of patients from group 1, 2 and 3, respectively. 98.7% of 388 patients avoided suprapubic fistula and resumed natural urination. Only 1.3% of patients were discharged with cystostomy drainage of the bladder because of urethral stricture, urethrorectal fistula, renal failure and pyelonephritis. The differential approach provided good outcomes in 87.6% of seriously ill patients in overall hospital lethality 8.9%. The majority of the lethal outcomes resulted from purulent pyelonephritis (71%). The rest cases were due to acute affections of circulation, respiration and digestion. PMID- 7474828 TI - [Use of hemodiafiltration in the treatment of patients with uremia]. AB - 21 patients on programmed hemodialysis for chronic renal insufficiency underwent hemodiafiltration (HDF) using dialyzers with highly-penetrable membrane made of polysulfone (Hemoflow F-40 and Hemoflow F-60). HDF has improved the patients' condition, eradicated skin itch, reduced polyneuropathy. Compared to standard hemodialysis, blood levels of creatinine and urea dropped by 6.9 and 3.7% more significantly. beta 2-microglobulin in the blood went down by 25.4 and 42.4%. The technique is provided of the removed filtrate replacement at the expense of the dialyzing liquid. The technique warrants a 60% decrease per session in blood beta 2-microglobulin. PMID- 7474829 TI - [Errors in the diagnosis and treatment of hormonally active pancreatic tumors]. PMID- 7474830 TI - [Clinical significance of alpha-1 acid glycoprotein and antiprotease blood proteins in salmonellosis and food poisoning]. PMID- 7474831 TI - [Effects of solar activity on clinical pathology]. PMID- 7474832 TI - [Effects of erythrocytopheresis on the state of the left heart in patients with polycythemia vera: echocardiographic findings]. AB - Functional response of the left ventricle (LV) and left atrium to erythrocytapheresis (EA) was studied in 20 patients. At admission, the patients had enlarged LV and low myocardial contractility. By cardiac and stroke volume indices two types of central hemodynamics were identified (hyperkinetic and hypokinetic) dependent on PV duration and concomitant cardiac pathology. These correlations were inferred basing on comparisons of echo-CG findings to data on the patients' age, sex, PV history, associated heart disease, packed cell volume. EA produced positive changes in echo-CG indices with complete normalization of cardiac output in some cases. This is supposed to be due to EA ability to limit foci of myocardial damage and to mobilize cardiac reserves. PMID- 7474833 TI - [Clinical features of gluten enteropathy]. PMID- 7474834 TI - [A case of Madelung's syndrome]. PMID- 7474835 TI - [Garcin's syndrome in a patient with malignant chemodectoma of the neck]. PMID- 7474836 TI - [A case of arrhythmogenic effects of quinidine, digoxin and finoptin combination]. PMID- 7474837 TI - [A complex of breathing exercises (hatha yoga) to arrest the developing attacks of dyspnea in bronchial asthma]. PMID- 7474838 TI - [Comparative study of the effectiveness of immunomodulating preparations in combined therapy of chronic obstructive bronchitis]. PMID- 7474840 TI - [Errors in the diagnosis of perforating gastroduodenal ulcers in the prehospital stage]. PMID- 7474839 TI - [Use of a new hemostatic preparation kaprofer in therapeutic endoscopy]. PMID- 7474841 TI - [Evaluation of work capacity of patients with histiocytosis X]. PMID- 7474842 TI - [Comparative effectiveness of anthelmintic preparations in trichocephaliasis]. PMID- 7474843 TI - [Problems of non-healing stomach ulcers: their classification and treatment]. PMID- 7474844 TI - [Effects of traditional antiulcer therapy on the motor-secretory function and microcirculation in the gastric and duodenal wall in patients with duodenal ulcer]. PMID- 7474845 TI - [Chrono-pharmacodynamic effects of sinepres in patients with hypertension]. PMID- 7474846 TI - [Clinical significance of detecting discrete antigens of group A Streptococcus and their antibodies in the blood of erysipelas patients]. PMID- 7474847 TI - [Dependence of long-term results of suturing perforating gastroduodenal ulcers on the occupation of patients]. PMID- 7474848 TI - [Effects of changes in central hemodynamics and hormonal systems of its regulation on the etiology of portal hypertension complications in patients with liver cirrhosis]. PMID- 7474849 TI - [Regulation of homeostasis in acute viral hepatitis B with protracted convalescence]. PMID- 7474850 TI - The effect of Antineoplaston, a new antitumor agent on malignant brain tumors. AB - Antineoplaston (Ap), a new antitumor agent, was clinically tested for effects on malignant brain tumors. The materials were 3 cases of glioblastoma (G,B), 2 cases of anaplastic astrocytoma, 1 case of pontine glioma, 2 cases of metastatic brain tumor and 1 case of medulloblastoma. All patients underwent radiochemotherapy and surgical resection of the tumors except the cases of pontine glioma, metastatic brain tumor and anaplastic astrocytoma. For gliomas, radiochemotherapy was used with Hu-IFN-beta. Ap was administered at a dose of 7-10 g/day in combination with remission maintenance therapy of gliomas. Complete response was obtained in one anaplastic astrocytoma. Partial response was obtained in 2 cases, a pontine glioma and a metastatic brain tumor. No change was observed in 2 cases, an anaplastic astrocytoma and a multiple brain metastasis. Progression of the disease was observed in 4 cases, 3 glioblastomas and 1 medulloblastoma, which showed continuous increase in tumor size. The effects of Ap on malignant brain tumors were considered due to synergy, since it was administered with other drugs and acceleration of tumor cellular differentiation. Ap is useful as an approach to remission maintenance therapy for brain tumors. PMID- 7474851 TI - Hormone receptors in pulmonary lymphangiomyomatosis. AB - Biochemical assays have demonstrated the existence of hormone receptors in lung tissue from patients with pulmonary lymphangiomyomatosis (LAM). This finding is the biologic corollary to the finding that LAM responds to hormonal therapy. We have examined lung tissue from two patients with LAM for estrogen and progesterone receptors by immunohistochemistry and a biochemical assay. Although specimens from both patients were negative for estrogen receptor activity by the biochemical assay, positive stain of estrogen receptors was observed in both patients with immunohistochemistry. PMID- 7474852 TI - Early mucosal gastric cancer with lymph node metastasis. AB - From 1976 to 1992, a total of 358 cases of mucosal early gastric cancer (m cancer) were treated by resection in the First Department of Surgery, Kurume University Hospital. Eight (2.2%) of these 358 cases of m-cancer were associated with lymph node metastasis. In these 358 cases, the mean diameter of the tumor was 2.75 cm, and the mean age was 59.6 years. In the 8 cases of m-cancer with positive lymph node metastasis (n(+)), the mean diameter was 5.2 cm and the mean age was 51.6 years. Seven (87.5%) out of the 8 cases of m-cancer with n(+) were IIc macroscopic cancer type and were associated with ulcer and/or ulcer scar. PMID- 7474853 TI - A novel adult T cell leukemia-derived cell line (SALT-3) susceptible to human immunodeficiency virus type 1 infection. AB - A novel human T cell line (SALT-3) was established from the pleural effusion of a patient with adult T cell leukemia (ATL) of lymphoma type. SALT-3 showed atypical T cell markers such as CD1-CD2-CD3-CD4+CD5+CD7+CD8-CD19-CD20-CD25+HLA-DR+. T cell receptor alpha/beta and gamma/delta were undetectable. Human T cell lymphotropic virus type 1 (HTLV-I) particles were seen on SALT-3 cells by electron microscopic analysis. HTLV-I gag p19, proviral DNA and mRNA of HTLV-I genes were also detected in the cells. Chromosome analysis showed abnormal karyotypes as 47, XY, partial trisomy of No.3 chromosome, and trisomy of No. 7 chromosome. Furthermore, SALT-3 were susceptible to the infection of human immunodeficiency virus type 1 (HIV-1) and the cells were rapidly killed after HIV-1 infection. This newly established HTLV-I-infected human T cell line would be a useful tool to study biological activities of atypical type of ATL cells and to examine the cytotoxic effects of HIV-1 and it's modulators. PMID- 7474854 TI - Blood flow velocity waveforms of the fetal middle cerebral artery in pregnancies complicated by diabetes mellitus. AB - Fetal blood flow velocity waveforms of the middle cerebral artery were measured by pulsed Doppler ultrasound in 43 pregnant women with diabetes mellitus (33 class B, 3 class C, 6 class D and 1 class R). The recordings were performed between 24 and 38 weeks of gestation. A total of 122 recordings were analyzed prior to establishing the resistance index (RI = peak systolic velocity minus diastolic velocity/peak systolic velocity). The mean maternal serum glucose was 124.3 mg/dl (range: 72.7 to 281.5 mg/dl), the mean hemoglobin A1c was 6.03% (range: 3.3 to 11.0%) and the mean fructosamine level was 255.97 mumol/L (range: 205 to 397 mumol/L). The resistance index did not differ between the fetuses of the diabetic mothers in our study population. Additionally, no significant correlation was noted between RI values and serum glucose levels (r = 0.03), hemoglobin A1c levels (r = 0.13) or fructosamine levels (r = 0.04) during the period of 26 to 34 gestational weeks. These data indicate that the RI within the fetal cerebral artery is unaffected by the maternal glycemic control when mean serum glucose levels are below 280 mg/dl. PMID- 7474856 TI - Murine monoclonal antibodies (MCS-1 and MCS-2) reactive with human myeloid leukemia cells. AB - In an attempt to identify the antigens expressed on human myeloid leukemia cells, two murine monoclonal antibodies (mAb) designated as MCS-1 (isotype; IgG3) and MCS-2 (IgG1) were raised. MCS-1 reacted with peripheral blood granulocytes, but not with monocytes, whereas MCS-2 reacted with both granulocytes and monocytes. The incidence of MCS-1 and MCS-2 reactivity with the cells from a total of 121 patients with various type of leukemias was as follows: 25/46 (54%) and 39/47 (83%) in acute myeloblastic leukemia and acute monoblastic leukemia, 8/16 and 16/16 in chronic myeloid leukemia in blastic crisis (CML-BC) of myeloid type, 0/7 and 2/7 in CML-BC of lymphoid type, 0/26 and 2/32 in acute lymphoblastic leukemia (ALL), 7/7 and 7/7 in chronic phase of CML, respectively. Two cases of MCS-2 positive ALL had a Philadelphia chromosome marker (Ph1). In contrast, neither MCS 1 nor MCS-2 reacted with lymphocytes or any of leukemic and non-leukemic lymphoid cell lines tested. These results indicate that MCS-1 and MCS-2 mAb recognized two different differentiation antigens expressed on granulocytes and monocytes. These mAb would be useful reagents to determine differentiation antigens on the cells in granulocyte and monocyte lineage. PMID- 7474855 TI - Neurokinin A depolarizes neurons of bullfrog dorsal root ganglia by suppressing K+ conductances. AB - The effect of neurokinin A (NKA) on neurons of bullfrog dorsal root ganglia (DRG) in primary culture was examined by using whole-cell patch-clamp methods. Application of NKA (1 microM) depolarized the DRG neurons, resulting in spontaneous firing of action potentials. Under voltage-clamp condition, NKA (3 nM 1 microM) caused an inward current (INKA) associated with decreased membrane conductance. The INKA reversed its polarity at the equilibrium potential for K+. The INKA was blocked by extracellular Ba2+ (1 mM) but not by nominally 0 mM Ca2+, tetraethylammonium (40 mM), 4-aminopyridine (2 mM) or apamin (50 nM). Intracellular Cs+ blocked the INKA. NKA depressed a voltage-dependent non inactivating K+ current, the M-current (IM), at potentials more positive than 55mV. NKA reduced the maximum M-conductance (GM) without changing the kinetics of M-channels. NKA also depressed a voltage- and time-independent background K+ current, IK(B). It is concluded that the INKA is produced by suppression of both IM and IK(B) in bullfrog primary afferent neurons. PMID- 7474857 TI - [A very delicate piece of work]. PMID- 7474858 TI - [The helping relationship with patients whose physical integrity is affected. Learning on site]. PMID- 7474859 TI - [The lessons of a voyage. Everyday life and health of the Mongol Nomads]. PMID- 7474860 TI - [Care and therapy of eczemas. So the itch remains bearable]. PMID- 7474861 TI - [The first interview as a basis for good relations with the patient. The decisive first interview]. PMID- 7474862 TI - [With music everything works better--or does it?]. PMID- 7474863 TI - [Considerations on a clinical experience. Evaluation at the patient's rhythm]. PMID- 7474864 TI - [SGGP conference: how much and which medicine do we need? The superfluous in medicine and care]. PMID- 7474865 TI - [How much care do we need? Care between too much and not enough]. PMID- 7474866 TI - [Sometimes we may use validation instead of valium]. PMID- 7474867 TI - [Professional politics. How are the professional interests represented?]. PMID- 7474868 TI - Hemorrhagic shock in rats. PMID- 7474869 TI - Mycobacterial infections of animals: pathology and pathogenesis. PMID- 7474870 TI - In vitro effects of Mycoplasma pulmonis on murine natural killer cell activity. AB - We evaluated the effect of coculture of virulent and avirulent strains of Mycoplasma pulmonis with mononuclear cells from resistant and susceptible strains of mice on natural killer (NK) cell activity against YAC-1 cells in a standard 4 h 51Cr-release assay. Endogenous NK activity was minimal in the specific-pathogen free-mice without an external stimulus. There was no correlation between in vivo virulence of the mycoplasmas or host resistance and the in vitro stimulation of NK cell activity. Only two of the avirulent strains of M. pulmonis tested induced significant increases in NK cell activity, and virulent M. pulmonis increased activity only in cells from susceptible C3H/HeN mice. PMID- 7474871 TI - Pancreatic adenocarcinoma in two grey collie dogs with cyclic hematopoiesis. AB - A 3-year-old male grey collie and 4-year-old female grey collie were part of a cyclic hematopoiesis study. Both dogs had experienced numerous bacterial infections, and both dogs were receiving various treatment regimens, including lithium and hematopoietic growth factors, to control the cyclic hematopoiesis. The first dog was presented in acute collapse and had a rapid clinical course. The second dog was presented with pyrexia and subsequently developed anorexia, disorientation, tremors, mild diarrhea, and bruising at venipuncture sites. Postmortem examination revealed pancreatic adenocarcinoma with metastasis in both cases. Pancreatic adenocarcinoma is a rare neoplasm in dogs. The incidence of pancreatic adenocarcinoma noted in this report is more than 150 times that previously reported in dogs. The cause of the increased incidence of pancreatic malignancy in these grey collies is unknown; possible factors include chronic inflammation or infections, chronic drug therapy, or genetic predisposition. Development of an uncommon neoplasm in two young grey collies may offer an opportunity to study the mechanisms of carcinogenesis. PMID- 7474872 TI - Thymoma in two ferrets. AB - A cranial mediastinal mass was observed radiographically in two 5-year-old adult male ferrets (Mustela putorius furo), both with histories of chronic episodic vomiting, dyspnea, and lethargy. Malignant lymphoma, a common neoplasm observed in ferrets, was considered the most likely diagnosis. Cardiomegaly and splenomegaly were also present. Histologically both mediastinal masses were composed of thymic epithelial cells and small lymphocytes, establishing a diagnosis of thymoma. Thymoma should now be included in the differential diagnosis of a cranial mediastinal mass in ferrets. PMID- 7474874 TI - Acute Mycoplasma pulmonis infection associated with coagulopathy in C3H/HeN mice. AB - Experimentally induced infection with high doses of Mycoplasma pulmonis results in acute pneumonia characterized by severe pulmonary hemorrhage, edema, and, often, death in C3H/HeN mice. To determine whether specific disease manifestations were associated with coagulopathy, we measured serum fibrin, fibrinogen degradation products, and plasma fibrinogen concentrations in C3H/HeN mice infected with high doses of a virulent strain of M. pulmonis. We also examined the lungs and other tissues from infected mice for the presence of intravascular fibrin clots and other lesions. Increased concentrations of fibrinogen degradation products indicated that coagulopathy occurs in acute M. pulmonis infection; however, intravascular fibrin clots were not present. Rather than decreasing, as might be expected during a consumptive coagulopathy, fibrinogen concentrations increased. The hyperfibrinogenemia probably is associated with an acute phase response to M. pulmonis infection. PMID- 7474873 TI - Acute pneumonia in a Syrian hamster: isolation of a Corynebacterium species. AB - A respiratory tract illness was detected in a 1-year-old male Syrian hamster; after it failed to respond to antibiotic therapy, the hamster was euthanized by CO2 administration. Postmortem examination revealed acute edematous pneumonia, and Corynebacterium paulometabulum was isolated from the lungs. PMID- 7474875 TI - Use of antibiotics to prevent hepatitis and typhlitis in male scid mice spontaneously infected with Helicobacter hepaticus. AB - The use of antibiotic combinations to prevent acute and progressive chronic hepatitis and proliferative typhlitis associated with Helicobacter hepaticus infection in male scid/NCr mice was evaluated. The drug combinations used were amoxicillin-metronidazole-bismuth, tetracycline-metronidazole-bismuth, amoxicillin-neomycin, neomycin alone, and amoxicillin alone. Treatments were administered per os for 14 days beginning at 4 weeks of age. All mice remained clinically normal throughout the study. Specimens from mice were evaluated histologically at 21, 60, 90, and 120 days after initiation of the antibiotic treatments. Results of histologic examination and use of special stains indicated that the antibiotic regimens containing amoxicillin prevented progressive chronic hepatitis and typhlitis. Helical bacteria were not observed histologically in the liver or cecum of amoxicillin-treated mice. Helical bacteria were observed in the liver and cecum of untreated mice and in the cecum of mice treated with antibiotic regimens not containing amoxicillin. Untreated mice and those treated with amoxicillin were evaluated by culture for presence of H. hepaticus at 60 and 90 days and by polymerase chain reaction at 90 days after initiation of the antibiotic treatment. All untreated mice were test-positive by fecal/cecal culture, and three of five were positive by polymerase chain reaction. All mice treated with amoxicillin were negative for H. hepaticus by results of culture and polymerase chain reaction. The oral administration of amoxicillin to young scid mice via the drinking water prevents hepatitis and typhlitis caused by H. hepaticus. PMID- 7474876 TI - Detection of Clostridium difficile toxins from the small intestine and cecum of rabbits with naturally acquired enterotoxemia. AB - Four specific-pathogen-free rabbits with anorexia died peracutely; decreased fecal output, nasal exudate, and labored breathing were the only other clinical abnormalities observed in two of the rabbits before death. The animals, three juveniles and one adult, were on a standard polyclonal antibody production regimen and had received immunizations approximately 2 weeks before presentation. External examination revealed distended abdomen and perineal fecal staining. At necropsy the small intestine was distended with fluid, and the cecum was distended with chyme. The small intestines and cecum had marked serosal hyperemia. Anaerobic bacterial culture techniques were used to isolate Clostridium difficile from the small intestine (3/4) and cecum (2/4). In all cases C. difficile toxin B was detected at high titers (10(2) to > 10(5)) in the small intestine by cytotoxicity assay with HeLa 229 cell culture. In two of the four rabbits C. difficile was isolated, and cytotoxin titers were detected at 10(1) and 10(4) in the cecum of affected rabbits. Toxin B was neutralized with C. sordellii antiserum but not C. spiroforme antiserum. In addition, toxin A was detected in each of the cytotoxin B-positive samples by a commercial toxin A enzyme immunosorbent assay. In vitro production of toxins A and B was detected from each culture isolate after incubation in chopped meat broth. These cases are noteworthy because spontaneous (nonantibiotic-associated) C. difficile enterotoxemia has not been previously reported in rabbits. Also the toxins of clostridial organisms are usually documented in the cecum, not the small intestine, of rabbits.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7474877 TI - New mutant rabbit strain with hypercholesterolemia and atherosclerotic lesions produced by serial inbreeding. AB - We have investigated the characteristics of the Kurosawa and Kusanagi hypercholesterolemic (KHC) rabbit, a strain manifesting inherited and persistent hypercholesterolemia and produced by inbreeding from mutants discovered in 1985. These KHC rabbits had abnormally high serum cholesterol, triglyceride, and phospholipid concentrations 8 to 10 times greater than the concentrations in clinically normal Japanese white rabbits (Oryctolagus cuniculus). The KHC rabbits also had decreased serum high-density lipoprotein cholesterol concentration, about one-third the value in clinically normal rabbits. In addition, the serum lipoprotein electrophoretic patterns were characterized by a strong, broad beta lipoprotein band and a diminished alpha-lipoprotein band. Fractionation of lipoprotein lipids revealed increased cholesterol, phospholipid, and triglyceride in the low-density lipoprotein fraction; increased cholesterol and phospholipid in the very low-density lipoprotein fraction; and decreased cholesterol and triglyceride in the high-density lipoprotein fraction. On the basis of incidence of hypercholesterolemia in mating experiments, this abnormality is thought to have been inherited as a single autosomal recessive gene mutation, and analysis of the low-density lipoprotein receptor indicated that the KHC rabbit has a 12 base pair deletion in the low-density lipoprotein receptor mRNA. Macroscopic analysis of the aorta revealed the atheromatous lesions at 2 months of age, drastically increased lesional areas in the total aortic surface at 8 months of age, and a high incidence of coronary atheromas and xanthomas. Light microscopy revealed two typical lesions in the aorta fatty plaque and medical calcification.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7474878 TI - Development of a rat model for iron deficiency and toxicological studies: comparison among Fischer 344, Wistar, and Sprague Dawley strains. AB - We carried out a comparative study to develop iron deficiency in three strains of rats: Fischer 344, Wistar, and Sprague Dawley. Considering that certain ingredients in the diet, such as sucrose, may inhibit enzymes of drug metabolism, which we planned to study during an iron deficiency state, the diet was modified to contain 35% each of sucrose and starch. Results of hematologic studies at the end of 6 weeks of iron deprivation clearly indicated that all clinical signs of iron deficiency anemia--hemoglobin concentration, protoporphyrin-to-heme ratio, and serum and liver iron concentrations--were affected in the three strains. However, the deficiency was less pronounced in Sprague Dawley than in Wistar and Fischer 344 strains. Results of this study suggest that rat strains respond differently to dietary iron inadequacy; this variability could be exploited to suit specific experimental needs. PMID- 7474879 TI - Development of a swine animal model for the study of sudden infant death syndrome. AB - An animal model with selective cardiac autonomic denervation was developed in neonatal swine for the purpose of future studies concerning the possible role of abnormal cardiac innervation in the etiology of sudden infant death syndrome. Twenty-six 4- to 8-day old piglets were randomly assigned to one of the following treatment groups: right stellate ganglionectomy (RSG), left stellate ganglionectomy (LSG), right cardiac vagotomy (RCV), and sham-operated control group. Piglets were anesthetized with Saffan and mechanically ventilated on room air. The thoracic wall was opened, and the sternum was divided, permitting exposure of pleura, heart, and lungs. After completion of one of these operative procedures, the thoracic wall was closed in layers. On successful recovery from anesthesia and extubation, piglets were returned to their sows. Postoperative administration of antibiotics was performed during the first week. Electrocardiograms were recorded once a week over an 8-week period. The sham operated control group had a postnatal increase in QTc and decrease in heart rate. The RSG group had significantly longer QTc than the sham-operated control group QTc, whereas the RCV group had significantly shorter QTc interval than sham operated controls. Surgical complications caused the death of two piglets (hemorrhage in one and aspiration in the other), infection caused the death of one piglet 2 weeks after surgery, and three died of unknown causes (one from the RSG group and two from the RCV group). The cause of death for these three animals could not be determined (necropsy was not revealing) because they were not monitored at the time of their demise.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7474880 TI - Use of surgical intrauterine insemination to manage infertility in a colony of research German shepherd dogs. AB - A colony of German shepherd dogs with inherited ventricular arrhythmias and sudden death had infertility that was resolved by intrauterine insemination. Mating of German shepherd males to related German shepherd females (40 cycles) by vaginal artificial insemination resulted in a low pregnancy rate of 35% and a small median litter size of 3.5. When these same German shepherd males were bred to female beagles by vaginal artificial insemination, the pregnancy rate of 100% (P = 0.02) and median litter size of 7.0 were significantly (P = 0.04) greater. Therefore, inadequate fertility existed when the German shepherds were mated. Because matings between these dogs were necessary, surgical intrauterine insemination of fresh semen was instituted in eight German shepherd females over nine cycles. In bypassing the cervix with this method, German shepherd fecundity increased significantly, with a pregnancy rate of 100% (P = 0.002) and median litter size of 8.0 (P = 0.001). Surgical intrauterine insemination may be an important method for management of canine infertility in the research environment. PMID- 7474881 TI - Retrospective evaluation of production characteristics in Sinclair miniature swine--44 years later. AB - Three hundred seventy-one litter records collected between 1985 and 1993 from 156 Sinclair S-1 miniature sows, a Hormel-derived strain of miniature swine, were retrospectively analyzed and compared with published records for 1950 to 1952 and 1963 to 1965. The effect of several variables such as season and month of parturition, age of sow, parity, and litter size on reproductive parameters of the Sinclair miniature swine were evaluated. The mean and standard error of the mean for litter size, number of liveborn, number of stillborn, and litter size at weaning of the Sinclair S-1 miniature swine were 7.20 +/- 0.12, 6.57 +/- 0.12, 0.63 +/- 0.06, and 5.75 +/- 0.12 piglets respectively. From a total of 2,436 liveborn piglets, 2,133 (87.56%) were weaned. The litter size at birth was similar to that previously reported for this strain of swine, whereas the litter size at weaning increased (P < 0.001) from 4.7 piglets during 1963 to 1965 to 5.8 piglets in our study. The average birth weight decreased (P < 0.001) from 0.90 kg in 1950 to 1952 and 0.72 kg in 1963 to 1965 to 0.59 kg in our study. Reproductive variables that had a considerable effect on the miniswine reproduction included month of parturition, age of sow, parity, and litter size. Primiparous sows had the smallest litter size and lowest number of weaned piglets. Sows during their second and third parity or sows between 2 and 4 years old had the best reproductive performances. Litter size and number of stillborn increased with parity and age of sow, resulting in fewer piglets weaned in older sows.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7474882 TI - Tissue preparation technique for microsphere assays of blood flow. AB - A simple method for preparing standardized tissue samples for microsphere assay of tissue blood flow is described. By reduction of tissue samples into a liquid state and centrifugation of the microspheres to the bottom of conical-shaped counting vials, the microspheres within all tissue and blood reference samples attain the same counting geometry. The validity and reproducibility of this technique has been established for planar-configured gamma-ray detectors in an in situ porcine kidney model. A positive correlation coefficient of r = 0.99 was observed between the total kidney blood flow, as measured by a calibrated Transonic flow probe, and the microsphere reference sample technique. PMID- 7474884 TI - Cardiopulmonary bypass with circulatory arrest in swine: echocardiographic evaluation of left ventricle function and pulmonary vein flow. AB - Complete two-dimensional echocardiography and Doppler flow studies were performed on 17 swine as part of an experimental protocol involving cardiopulmonary bypass with circulatory arrest. Indices of left ventricular function and pulmonary vein morphology and flow velocity were taken before (17/17) and after bypass (11/17). A total of 70 measurements were available for analysis. Doppler flow velocity within the right pulmonary vein was 0.5 +/- 0.06 m/sec and was 0.52 +/- 0.05 m/sec in the left pulmonary vein. The swine were found to have a marked reduction in left ventricle function compared with that in previous reports, though no statistical difference was seen between pre- and post-cardiopulmonary bypass studies (preoperative shortening fraction was 21 +/- 5% and ejection fraction was 41 +/- 8%; postoperative shortening fraction was 23 +/- 5% and ejection fraction was 42 +/- 7%). Swine with shortening and ejection fractions of < 20% were found to have more unstable hemodynamics during cardiopulmonary bypass. PMID- 7474885 TI - Diagnostic exercise: ataxia and incoordination in ferrets. PMID- 7474886 TI - Case report of listerial keratoconjunctivitis in hairless guinea pigs. PMID- 7474887 TI - Retrospective observation of the effect of accidental exposure to organophosphates on the success of embryo transfer in mice. PMID- 7474883 TI - Comparison of the effects of five adjuvants on the antibody response to influenza virus antigen in guinea pigs. AB - Five adjuvants were tested for their effect on the immune response in guinea pigs to the hemagglutinin antigen of influenza virus strain B/Panama. Vaccines containing 924 micrograms of hemagglutinin antigen/ml were prepared at high and low doses of Freund's complete and incomplete adjuvants, Syntex adjuvant, RIBI's adjuvant, TiterMax adjuvant, and aluminum phosphate adjuvant. Responses to these vaccines were compared with those to a control vaccine containing influenza virus B/Panama hemagglutinin antigen and saline. On day 28, vaccines containing the following adjuvant doses had significantly higher titers than the titer for the control: Freund adjuvants at high and low doses, RIBI at high dose, TiterMax at high and low doses, and aluminum phosphate at high dose. On day 42, vaccines containing the following adjuvant doses had significantly higher titers than that for the control: Freund adjuvants at high and low doses, RIBI at high dose, TiterMax at high dose, and aluminum phosphate at high dose. Freund adjuvants at high and low doses, RIBI adjuvant at high dose, and aluminum phosphate at high dose caused significantly greater swelling at the inoculation site than did the control vaccine. TiterMax adjuvant at high and low doses, and aluminum phosphate at low dose caused minor swelling at the inoculation site, but it was not significantly different from the swelling caused by the control vaccine. Syntex adjuvant at high and low doses, RIBI at low dose, and control (saline/antigen) at high and low doses caused no swelling after inoculation. Overall, the high dose of adjuvants caused greater tissue swelling than did the low dose of adjuvants.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7474888 TI - Urinary calculus caused by plant material in a cynomolgus monkey. PMID- 7474889 TI - Development of ocular hypertension in congenitally buphthalmic rabbits. PMID- 7474890 TI - Pathogenesis of acute necrotic hepatitis in rabbit hemorrhagic disease. PMID- 7474891 TI - Lipid status and fatty acid metabolism in phenylketonuria. AB - Owing to dietary treatment, patients with phenylketonuria (PKU) are supplied with only small amounts of animal fats and show low blood cholesterol levels and a poor long-chain polyunsaturated fatty acid (LC-PUFA) status. Endogenous synthesis should compensate for the physiological need for cholesterol for growth, but supplementary exogenous LC-PUFA seems advisable. PKU infants could develop depletion of docosahexaenoic acid, fundamental for the functional development of the central nervous system. The availability of arachidonic acid from plasma in PKU has been found to be related to dietary compliance, and seems to influence the synthesis of arachidonate-derived eicosanoids. Trials of dietary supplementation of PKU patients with oils containing LC-PUFA are in progress, but the functional consequences of these supplementations need further investigation. PMID- 7474892 TI - Identification of two new aberrant splicings in the ornithine carbamoyltransferase (OCT) gene in two patients with early and late onset OCT deficiency. AB - Ornithine carbamoyltransferase (OCT) is a liver-specific enzyme located in the mitochondrial matrix. OCT deficiency is an X-linked disease with a heterogeneous phenotype, even in affected males. We studied two male patients (K.M., K.G.) with early and late onset, respectively. OCT activity was zero in the autopsied liver of patient K.M. and was 6% of control in the biopsied liver of K.G. Sequencing of OCT cDNAs revealed exon 5 skipping in K.M., resulting from a T-to-C transition of the initial dinucleotide of the 5' splicing donor site of intron 5, and a G-to-T transversion at position +45 in exon 9 (L304F) in K.G., providing three OCT mRNAs of different lengths: a normally spliced transcript, 23 bp insertion of intron 8 and the first 50bp missing within exon 9. Exon 5 skipping and two other aberrant splicings produced stop codons early downstream in mature OCT mRNAs. Expression study of a missense allele, L304F, transfected to cultured Cos 1 cells revealed a 34.4% value of the control. The difference of OCT activities between the patient liver and transfected cells (6% vs. 34%) can be explained by this splicing abnormality. PMID- 7474893 TI - Defective molybdopterin biosynthesis: clinical heterogeneity associated with molybdenum cofactor deficiency. AB - A patient with molybdenum cofactor deficiency (producing the biochemical abnormalities associated with deficiencies of sulphite oxidase and xanthine dehydrogenase) clinically expressed Marfan-like habitus with dislocated lenses, vertebral abnormality, learning disability, moderate hemiplegia, increased medial lentiform MRI signal and intermittent microscopic haematuria. S-Sulphocysteine was present in plasma and urine, and the oxidized derivative of a molybdopterin precursor (precursor Z), together with xanthine and hypoxanthine, were elevated in urine. Blood uric acid was < 1 mg/dl, while urinary urothione was not detected. These data indicate a functionally inadequate terminal enzyme for converting precursor Z to active molybdopterin (complementation group B of general molybdenum cofactor deficiency). Although the biochemical parameters were indicative of a severe deficiency state, the patient has survived into the third decade with a less severe clinical spectrum than has generally been associated with this disease. PMID- 7474894 TI - A new diagnostic technique for adenylosuccinate lyase deficiency. AB - A convenient and simple method of diagnosing adenylosuccinate lyase (ASL) deficiency is described. This method consists of (1) isolation of SAICA riboside and S-Ado with a cation exchange resin; (2) measurement of the UV absorbance of the ammonia eluate at 270 and 250 nm; (3) calculation of the A270/A250 ratio. If the value of this ratio is less than 0.45, the patient has a normal level of ASL activity. If the value of this ratio is greater than 0.70, the patient has ASL deficiency. PMID- 7474895 TI - Prenatal diagnosis and therapy for a patient with vitamin B12-responsive methylmalonic acidaemia. AB - The prenatal therapy is described of a patient with vitamin B12-responsive methylmalonic acidaemia during the last 10 days of gestation with oral administration of vitamin B12 (20 mg/day) given to a mother did not normalize her urinary excretion of methylmalonic acid (MMA), which was 14.5 mmol/mol creatinine at 32 weeks of gestation. Before delivery, the mother was excreting 18.9 +/- 3.3 mmol MMA/mol creatinine (mean value at 7 days after vitamin B12 therapy), as well as at 32-37 weeks of gestation with no therapy. After birth, the level of MMA in the infant's urine was remarkably elevated (500-700 mmol/mol creatinine); the level of MMA in maternal urine decreased dramatically after delivery. Compared with two previous reports, the length of administration was not sufficient to reduce maternal MMA excretion. In future, the length of the therapy, route of administration and total dose of vitamin B12 to maintain an efficient level of vitamin B12 in an affected fetus should be considered. PMID- 7474896 TI - Glycine and L-carnitine therapy in 3-methylcrotonyl-CoA carboxylase deficiency. AB - Genetic deficiency of 3-methylcrotonyl-CoA carboxylase (3-MCC) is a rare inborn error of leucine metabolism producing an organic acidaemia. With accumulation of 3-methylcrotonyl-CoA, there is increased production of 3-hydroxyisovaleric acid, the glycine conjugate (3-methylcrotonylglycine), and the carnitine conjugate (3 hydroxyisovalerylcarnitine). The conjugates represent endogenous detoxification products. We studied excretion rates of these conjugates at baseline and with glycine and carnitine therapy in an 8-year-old girl with 3-MCC deficiency. Her preadmission diet was continued. Plasma and urine samples were obtained after 24 h of each of the following: L-carnitine 100 mg/kg per day and glycine 100, 175 and 250 mg/kg per day. Plasma and urinary carnitine levels were reduced by 80% and 50%, respectively with abnormal urinary excretion patterns. These normalized with carnitine therapy. Acylcarnitine excretion increased with carnitine therapy. The glycine conjugate, 3-methylcrotonylglycine (3-MCG), was the major metabolite excreted at all times and its excretion increased with glycine therapy. Clearly, in 3-MCC deficiency the available glycine and carnitine pools are not sufficient to meet the potential for conjugation of accumulated metabolites, suggesting a possible therapeutic role for glycine and carnitine therapy in this disorder. PMID- 7474897 TI - Trimethylaminuria, fish odour syndrome: a new method of detection and response to treatment with metronidazole. AB - Trimethylaminuria is an autosomal recessive disorder involving deficient N oxidation of the dietary-derived amine trimethylamine (TMA). TMA, a volatile tertiary amine, accumulates and is excreted in urine of patients with deficient TMA oxidase activity. Treatment strategies for this condition are limited. We report a new stable-isotope dilution method for rapid sequential analysis of TMA concentrations and the clinical and biochemical response to treatment with metronidazole. PMID- 7474898 TI - Heterozygotes for plasmalemmal carnitine transporter defect are at increased risk for valproic acid-associated impairment of carnitine uptake in cultured human skin fibroblasts. AB - One of the mechanisms by which chronic valproic acid (VPA) therapy induces serum and tissue depletion of carnitine in normal controls is through inhibition of plasmalemmal carnitine uptake (Tein et al 1993). To determine the effect of VPA on proven heterozygotes for the plasmalemmal carnitine transporter defect, we studied this system in cultured human skin fibroblasts with reduced Vmax for the carnitine transporter using L-[3H]carnitine. There was en exponential dose dependent decrease in carnitine uptake with increasing VPA concentrations and the relative inhibitory effect was the same for all three carnitine concentrations for a given cell line. Importantly, the lower the maximal velocity of carnitine uptake of the heterozygote, the lower the number of carnitine transporters and the lower the carnitine uptake per given concentration of VPA. The degree of inhibition was also directly proportional to the time of VPA preincubation up to a specific maximal saturation time. The maximal effect of VPA exposure time was reached by 10 days in the control cell line and by 3 days in the two heterozygote lines, probably reflecting earlier saturation. We conclude that patients who are heterozygous for the plasmalemmal carnitine transporter defect are at increased risk for VPA-associated serum and tissue depletion of carnitine through inhibition of plasmalemmal carnitine uptake. PMID- 7474899 TI - A new Japanese case of succinyl-CoA: 3-ketoacid CoA-transferase deficiency. PMID- 7474900 TI - Metachromatic leukodystrophy among southern Alaskan Eskimos: molecular and genetic studies. AB - Metachromatic leukodystrophy (MLD) is an autosomal recessive disorder resulting from the inability to metabolize sulphatide, an important component of myelin. Although there is significant clinical variability between patients, most have the late-infantile form. It is one of the most common lysosomal disorders involving mental deterioration and is found throughout the world. The great majority of the cases have a deficiency of arylsulphatase A activity. Accurate diagnosis of MLD is complicated by the presence of so-called pseudodeficiency alleles and the need to receive specimens for biochemical testing within 24-48 h of collection. We report the identification of the mutation (a g-to-a transition in the first nucleotide of intron 4) in the arylsulphatase A gene causing late infantile MLD among the Eskimo population of southern Alaska. As all patients and family members from living and deceased patients had the same mutation, a mutation-based test was developed to identify patients and carriers that can be done on dried blood spots sent via regular mail service. A possible genetic link between this population and the Navajo Indians of the southwestern United States is proposed. PMID- 7474901 TI - Familial isolated vitamin E deficiency. Extensive study of a large family with a 5-year therapeutic follow-up. AB - A major neurological deterioration, beginning with ataxia, led to the diagnosis of familial vitamin E deficiency in a girl. Based upon vitamin E determinations, 4/8 members of the (consanguineous) sibship were considered to be homozygous. Homozygosity was also found for the alleles of six markers linked to the AVED locus, recently identified in similar Tunisian or Sicilian families on chromosome 8q. Measures of vitamin E in lipoprotein fractions and in liver biopsy after vitamin E oral load suggested that free diffusion of vitamin E between the different compartments was possible and even increased. However, a high-affinity ligand seemed to be lacking, either in the hepatic recycling of vitamin E or in both the hepatic and the other vitamin E compartments. The 5-year substitutive treatment was successful only in the pre- or paucisymptomatic patients. Serum vitamin E must be measured in any unexplained progressive ataxia. PMID- 7474902 TI - Inborn errors of metabolism in Singapore. AB - Between 1992 and 1994 the Paediatric Metabolic Centre screened 403 patients for inherited metabolic disease. Seventeen patients were diagnosed, including 7 with organic acidurias, 5 with mucopolysaccharidoses, 2 with amino acid disorders, 2 with carbohydrate disorders and 1 with hyperlipidaemia. The ethnic distribution of disorders in Singapore is compared with incidences reported by others in this part of the world. It is likely that further studies will indicate which diagnostic or screening tests would best serve this community. PMID- 7474904 TI - Immunological abnormalities in a patient with tyrosinaemia type III. PMID- 7474903 TI - Phenylketonuric patients decades after diet. AB - Nineteen early-treated phenylketonuric patients, whose diet was discontinued between 4.5 and 13 years of age, and who have been off the diet for 12-28 years, were reassessed in 1992-93. There was little change in mean IQ between end of diet and follow-up, less than one IQ point on the average, with no change for any individual exceeding 12 IQ points. Both prior and current IQ correlated slightly negatively with mean phenylalanine (Phe) concentration, and positively with parents' education. The phenylalanine level at follow-up was significantly lower on average by about 900 mumol/L. Five of the subjects (26%) have evidence of mental disease. However, the data suggest that the discontinuation of the diet did not cause intellectual deterioration. Nonetheless, the patients' intellect cannot be the only consideration for maintenance of diet. The occurrence of psychopathology among phenylketonuric patients and the possible unknown effects of toxic elevation of phenylalanine during their lifetime suggest the need to maintain the diet. The use of DNA for diagnostic and prognostic purposes might assist in decisions about dietary quality and duration, and in anticipation of psychopathology. PMID- 7474905 TI - A novel point mutation at codon 269 of the ornithine transcarbamylase (OTC) gene causing neonatal onset of OTC deficiency. PMID- 7474906 TI - Glutaric aciduria mediated by gut bacteria. PMID- 7474907 TI - Neuronal ceroid lipofuscinosis with prominent accumulation of ceroid lipofuscin in the myocardium. PMID- 7474908 TI - Progressive pulmonary hypertension: a fatal complication of type I glycogen storage disease. PMID- 7474909 TI - Infantile form of carnitine palmitoyltransferase II deficiency in a girl with rapid fatal onset. PMID- 7474910 TI - Prenatal diagnosis of Menkes disease by mutation analysis. PMID- 7474911 TI - Medium-chain acyl-CoA dehydrogenase deficiency is not a cause of previously diagnosed Reye syndrome. PMID- 7474912 TI - Organ donation by a maple syrup urine disease patient. PMID- 7474913 TI - Mutational analysis of Turkish galactosaemia patients. PMID- 7474914 TI - Transient neonatal hyperphenylalaninaemia is not related to mutations at the phenylalanine hydroxylase gene. PMID- 7474915 TI - Apolipoprotein E and amyloidogenesis. PMID- 7474916 TI - The renal toxicity of sustained glomerular protein traffic. PMID- 7474917 TI - Association of apolipoprotein E with murine amyloid A protein amyloid. AB - BACKGROUND: Experimental amyloid A protein (AA) amyloidosis in mice is the most rapid type of amyloid formed, thus providing a valuable model to study amyloid formation. Recent studies have suggested the importance of apolipoprotein E (apoE) in Alzheimer's disease and systemic amyloidoses. To help understand the role of apoE in amyloidoses, we examined amyloid tissue for the presence of apoE in mouse AA amyloid. EXPERIMENTAL DESIGN: Mice were injected with amyloid enhancing factor and silver nitrate to induce amyloid deposition. Spleens were examined for Congo red staining and serum amyloid A, serum amyloid P component, and apoE immunostaining. In addition, RNA analysis was performed to measure the expression of apoE in various tissues after amyloid induction. RESULTS: We have found that apoE is associated with mouse amyloid. Ab to apoE consistently detected the presence of increased levels of apoE in amyloid tissue. Immunohistochemical analyses confirmed that the apoE immunoreactivity co associated with AA and serum amyloid P in the amyloid fibrils. Northern blot analysis of amyloid tissue showed an increase in apoE messenger RNA compared with control tissue. CONCLUSIONS: This is the first demonstration of apoE in mouse amyloid tissue. The data presented here, along with previous studies, suggest that apoE may be involved in amyloidogenesis. These studies validate the mouse model for studying the role of apoE in amyloid fibrillogenesis. The use of transgenic and gene-inactivated mice will help to elucidate the role and mechanism of apoE in amyloid formation. PMID- 7474918 TI - Identification of lysyl oxidase and other platelet-derived growth factor inducible genes in vascular smooth muscle cells by differential screening. AB - BACKGROUND: Atherosclerosis and arterial injury are characterized by vascular smooth muscle cell (VSMC) migration and growth and an increase in synthesis of extracellular matrix. Platelet-derived growth factor (PDGF) has been implicated in these processes. This study was designed to identify additional PDGF-regulated genes in VSMC. EXPERIMENTAL DESIGN: A cDNA library prepared from PDGF-stimulated rat aortic VSMC was screened by differential hybridization to identify clones representing PDGF-inducible genes. The time course of growth factor-induced changes in gene expression was examined by RNA blot hybridization. Assays of protein activity were also performed for selected gene products. RESULTS: Four PDGF-regulated cDNA clones were identified by DNA sequencing. These encoded the extracellular matrix proteins lysyl oxidase (LO), thrombospondin, and osteopontin and the intracellular enzyme lactate dehydrogenase (LDH). Levels of mRNA corresponding to all four genes were low in quiescent VSMC and were markedly induced by PDGF, angiotensin II, and 10% calf serum. The regulation of LO and LDH mRNA by these agonists in VSMC has not been previously reported. LO enzymatic activity in the culture media was increased by approximately equals to 700% after exposure to PDGF. In contrast, LDH activity was not increased by PDGF treatment. CONCLUSIONS: The induction of LO mRNA and its secretion by VSMC is an early event accompanying growth factor stimulation and may contribute to organization of the extracellular matrix. PMID- 7474919 TI - Expression of the hepatocyte growth factor/scatter factor receptor tyrosine kinase is localized to epithelia in the adult mouse. AB - The met proto-oncogene receptor tyrosine kinase has been identified as a receptor for hepatocyte growth factor (HGF)/scatter factor (SF). HGF/SF is a multifunctional cytokine that stimulates mitogenesis, dissociation, and motility of a broad spectrum of epithelial and endothelial cells in culture, promotes the progression of carcinoma cells to a more invasive phenotype, and acts as a morphogenic factor for tubular epithelia. HGF/SF is predominantly expressed by mesenchymal cells, whereas the met/HGF/SFR is predominantly expressed by epithelial and carcinoma cells in culture. We have shown by Northern analyses that the met/HGF/SFR is expressed in many adult mouse tissues. To elucidate the normal physiologic role for the met/HGF/SFR and the possible pathologic consequences of deregulation of this pathway, we have examined the expression of the met/HGF/SFR in adult mouse tissue by in situ hybridization. We show that the met/HGF/SFR is generally expressed in epithelia, including hepatocytes, epithelial cells that line the proximal and distal convoluted tubules of the kidney, epithelia of stomach, esophagus, uterus, lung and skin, as well as in granulosa cells of developing and mature oocytes. By reverse transcriptase PCR amplification, we show that the HGF/SF gene is expressed at low levels in many of these tissues. Our data support a possible role for the met/HGF/SFR in epithelial cell growth and tissue organization. PMID- 7474920 TI - Hodgkin cells accumulate mRNA for bcl-2. AB - BACKGROUND: The bcl-2 oncogene is able to prevent cells from apoptosis. Overexpression of the bcl-2 protein seems to be important for the pathogenesis of follicular center cell lymphomas, in which both protein and mRNA usually show high levels. In addition, the expression of the Epstein-Barr virus-encoded late membrane protein up-regulates the bcl-2 protein in cell lines. The aim of the current study was to investigate the expression of the bcl-2 oncogene in Hodgkin's disease both at the protein and mRNA level in correlation with the expression of the late membrane protein. EXPERIMENTAL DESIGN: Thirteen cases of all histologic types of Hodgkin's disease, six cases of chronic nonspecific lymphadenitis, three tonsils with follicular hyperplasia, seven cases of follicular small cleaved cell lymphoma, and six cases of follicular large cell lymphoma, were analyzed. We designed a novel digoxigenin-labeled oligonucleotide probe complementary to bcl-2 mRNA for nonisotopic in situ hybridization. Bcl-2 oncoprotein and late membrane protein expression were determined by immunohistochemistry. The presence of the 14;18 translocation was analyzed by PCR for the major breakpoint region. RESULTS: The main finding was that, irrespective of subtype, the vast majority of Hodgkin cells express abundant bcl-2 mRNA. Oncoprotein expression, however, varied from case to case, with the highest prevalence in the nodular sclerosing subtype, and showed no strict correlation with the late membrane protein. In our case, no 14;18 translocation could be found in Hodgkin's disease. CONCLUSIONS: Hodgkin cells in all types of Hodgkin's disease demonstrated high levels of bcl-2 mRNA, while the bcl-2 protein expression was inhomogenous. In nodular lymphocyte predominant type, the bcl-2 mRNA and protein pattern is comparable to germinal center cells. This finding is a further argument for the germinal center cell origin of this type of Hodgkin's disease. PMID- 7474921 TI - The quantity of nucleolar proteins nucleolin and protein B23 is related to cell doubling time in human cancer cells. AB - BACKGROUND: The quantity of the silver-stained nucleolar proteins (AgNOR proteins) measured in situ in cytohistologic preparations is related to the rapidity of cell proliferation. The term "AgNOR proteins" comprises several proteins. The relationship between the individual AgNOR protein amount and cell proliferating activity is not yet known. We studied the quantitative distribution of the individual AgNOR proteins, with specific attention to the two major AgNOR proteins, nucleolin and protein B23, in seven human cancer cell lines characterized by different cell doubling times. DESIGN: The doubling time of cancer cells was measured by counting the asynchronously growing cells at regular time intervals. The AgNOR proteins were quantified in situ, after a specific one step staining procedure, by computerized image analysis. For the quantitative evaluation of nucleolin and protein B23, two methods were followed. Nuclear proteins after separation by SDS-PAGE were transferred onto nitrocellulose membranes and were either: 1) stained by the silver staining procedure for AgNOR proteins or 2) treated with anti-nucleolin and anti-protein B23 mAb followed by reaction with secondary Ab linked to peroxidase and revealed by chemiluminescence and autoradiography. In both cases, measurement of individual AgNOR protein and nucleolin and protein B23 amount was carried out using computerized densitometric analysis. RESULTS: Integrated density values of the silver-stained bands at 105 kDa (nucleolin) and 38 to 39 kDa (protein B23) represented, in all cell lines, more than 60% of the total silver-stained band value. A relationship was found between the densitometric values of silver-stained nucleolin and protein B23 and rapidity of cell proliferation (r = 0.85 and r = 0.86, respectively, p < 0.05). The values of nucleolin and protein B23 obtained using the Western blots were strictly related to the rapidity of cell proliferation (r = 0.93 and 0.96, respectively, p < 0.001). Finally, a good correlation was observed between the mean AgNOR protein area value, as defined in cytologic preparations in situ, and nucleolin and protein B23 amounts as evaluated in silver-stained nitrocellulose membranes (r = 0.92 and r = 0.90, respectively, p < 0.01) and in Western blots (r = 0.95 and r = 0.94, respectively, p < 0.001). CONCLUSIONS: These data indicate that the quantitative changes of AgNOR proteins observed in cytohistologic preparations in situ mainly reflect the quantitative changes of nucleolin and protein B23 and demonstrate that nucleolin and protein B23 amounts are inversely related to cell doubling time in human cancer cells. PMID- 7474922 TI - Second primary head and neck squamous cell carcinoma predicted by the glutathione S-transferase expression in healthy tissue in the direct vicinity of the first tumor. AB - BACKGROUND: Glutathione S-transferases (GST) are known to play a role in the detoxification of carcinogens. Individual isoenzymes of the alpha-, mu-, and pi class vary in substrate specificities, tissue distribution, and activities among individuals. GST-pi expression has been shown to be increased in preneoplastic and neoplastic lesions. GST-mu is known to play a role in detoxification of epoxides released from cigarette smoke, and individuals with low GST-mu activity have a relatively high risk to develop smoking-related lung and laryngeal cancer. The occurrence of a second primary tumor (SPT) in the whole respiratory and upper aerodigestive tract is an important factor for mortality in head and neck squamous cell carcinoma (HNSCC), and, at present, there are no markers that are available to predict which patient has increased chances of developing an SPT. Risk-assessment by use of biomarkers, particularly the ones that can be obtained with noninvasive techniques, are of great value in predicting prognosis and hence possibly more aggressive treatment and follow-up in selected patient groups. EXPERIMENTAL DESIGN: In a nested case control study, 20 patients who had previous history of oral cancer were used; 10 of the 20 had developed an SPT, and the other 10 patients were minimally 7 years free of disease. The expression of GST pi, GST-mu, and GST-alpha was immunohistochemically analyzed using apparently normal oral mucosa, free of tumor or dysplasia, obtained from the resection edges around the primary tumor. In another experiment, the three GST isoenzymes were immunohistochemically analyzed using exfoliated cells, obtained noninvasively from several sites of the upper aerodigestive tract of the apparently normal looking mucosa of HNSCC patients (n = 25) and of control individuals (n = 10). RESULTS: The expression of all GST was significantly higher (p < 0.001) in the suprabasal and superficial layers of the mucosa at risk. Also, in cell scrapes of clinically healthy mucosa of HNSCC patients, we observed a significantly higher expression (p < 0.001) of GST-pi and GST-mu compared with their matched controls. For GST-alpha, we observed a more heterogenous expression pattern in these exfoliated cells. CONCLUSIONS: Expression of GST-pi, -mu, and -alpha in normal tissue in the direct vicinity of the first tumor seems to have predictive value for the development of an SPT. PMID- 7474923 TI - Loss of follicular dendritic cells in murine-acquired immunodeficiency syndrome. AB - BACKGROUND: The disease caused by HIV-1 leads to the destruction of follicular dendritic cells (FDC) and the follicular architecture in secondary lymphoid tissues. The murine acquired immunodeficiency syndrome (MAIDS, caused by LP-BM5) serves as an animal model for study of mechanisms involved in development of retrovirus-induced immunodeficiencies. The present study was undertaken to determine whether LP-BM5 infection leads to the destruction of murine FDC and the normal follicular architecture in secondary lymphoid tissues. EXPERIMENTAL DESIGN: Mice were infected with LP-BM5, and the follicular architecture and FDC networks were assessed. The pathologic changes observed were correlated with FDC function. RESULTS: Three weeks after infection, FDC networks were present, and they often appeared hyperplastic. However, by 1 month after infection, distorted lymphoid follicles were apparent, and the intensity of FDC labeling began to decline. FDC disappeared first in the spleen, and in hyperimmunized mice, FDC in draining lymph nodes disappeared before FDC in nondraining lymph nodes. By 4 months, the normal follicular localization of B cells was missing, and FDC were not detectable in most tissues. As the FDC and the normal lymphoid architecture degenerated, extrafollicular foci of immunoblasts and plasma cells appeared in areas typically reserved for T cells, and the Thy 1.2+ T cells were dispersed. Of interest, the total number of Ig-producing cells increased as the disease progressed; in contrast, the number of anti-human serum albumin-producing cells in mice immunized with human serum albumin before infection decreased. CONCLUSIONS: These data indicate that, like HIV-1 infection, LP-BM5 infection leads to the loss of FDC and the normal follicular architecture. However, morphologic changes were not observed until after FDC had lost their normal ability to trap and retain Ag. These data indicate that retroviral infections may cause FDC dysfunctions long before FDC are destroyed. PMID- 7474924 TI - In situ behavior of human Langerhans cells in skin organ culture. AB - BACKGROUND: Epidermal Langerhans cells (ELC) play a critical role in the initiation of cutaneous immune responses. ELC are characterized by the expression of major histocompatibility complex (MHC) class II Ag and a number of adhesion/costimulatory molecules. Evidence suggests that cytokines induced within the epidermis regulate the functions of ELC, including their phenotypic expression. In the human system, no information is available regarding the behavior of the ELC in situ: their changes in morphology, expression of functional molecules or migration within the microenvironment. In the present study, using an ex vivo human skin organ culture model, we addressed the above questions and also examined the phenotypic modulation of ELC in situ by cytokines. EXPERIMENTAL DESIGN: Skin explants were cultured either in a Trowell type method or free in the medium. Skin explants were cultured with and without cytokines and were processed for light and electron microscopy and for immunohistochemical definition of ELC phenotypes. RESULTS: In the Trowell-type skin organ culture, morphologic integrity of ELC, CD1a molecule, and Birbeck granules could be preserved intact up to 3 to 4 days in culture. During the first 3 days of culture, the intensity of MHC-II (HLA-DR, DP, and DQ) and CD1a expression on ELC increased sharply, and the dendritic appearance of ELC became more prominent at Day 3. Adhesion molecules, ICAM-1, LFA-3, HECA-452, sLx, and B7/BB1 were also spontaneously acquired in varying amounts by CD1a+ ELC after 3 days in culture. Significant increase of CD1a and ICAM-1 expression on ELC was observed within 12 hours, when skin explants were cultured free in the medium with GM-CSF and TNF-alpha, respectively. Further, we demonstrated spontaneous migration of ELC within the epidermis and then to the dermis during the Trowell type skin culture. We also showed the migration of ELC out of the human skin when skin explants were cultured directly in the medium. CONCLUSIONS: Human ELC showed significant phenotypic changes within the epidermis and acquired migratory capacity during the skin organ culture. ELC in skin organ culture appear to undergo a phenotypic maturation within the epidermis. ELC in situ rapidly respond to GM-CSF and TNF-alpha by increasing the expression of CD1a and ICAM-1 molecules, respectively. These results suggest the modulation of phenotypic characteristics of ELC and their migration in response to the changes of epidermal microenvironment and cytokines and implicate the potential use of skin organ culture model to elucidate the role of human ELC in the immunopathology of skin diseases. PMID- 7474925 TI - Stromal cells from subcutaneous adipose tissue seeded in a native collagen/elastin dermal substitute reduce wound contraction in full thickness skin defects. AB - BACKGROUND: Dermal substitutes seeded with cultured fibroblasts have been developed to improve dermal regeneration in full thickness wounds. Because of cell cultivation, 3 weeks are required before patients can be treated with these autologous adipose tissue. This substitute is easily fabricated within hours, which allows immediate treatment of full thickness defects. EXPERIMENTAL DESIGN: Porcine full thickness wounds were substituted with native collagen/alpha-elastin hydrolysate matrices. One group of matrices was left unseeded as negative control. The second was seeded with cultured dermal fibroblasts as positive control. The third was seeded with a stromal-vascular-fraction of adipose tissue, and the fourth was seeded with a stromal fraction with few vascular fragments (SF). All substitutes were covered with split skin mesh grafts and were protected against dehydration and infection with a microporous polyether urethane membrane. For 8 weeks, weekly biopsies were taken, myofibroblasts and fibroblasts were counted, thickness of the granulation tissue band was measured, and wound contraction and histology were evaluated. RESULTS: Negative control and stromal vascular-fraction substitutes were invaded by high numbers of myofibroblasts and fibroblasts. They did not reduce wound contraction, and scar tissue was formed. SF substitutes reduced the accumulation of myofibroblasts and fibroblasts and prevented the formation of granulation tissue. As a result, dermal regeneration improved, and wound contraction was less than by the other substitutes. CONCLUSIONS: Adipose tissue cell isolates included vascular fragments containing endothelial cells. Seeded in dermal substitutes, these vascular fragments induced hypergranulation tissue formation and caused wound contraction. SF substitutes contained few endothelial cells. As a result, the contraction arresting effect of the seeded stromal cell fraction was effective. Our concept of a cellular dermal substitute seeded with stromal cells from adipose tissue is feasible and allows immediate treatment of full thickness skin defects. PMID- 7474926 TI - Suppressive effect of anti-alpha 1-antichymotrypsin serum on pulmonary fibrosis induced by phorbol myristate acetate in vivo. AB - BACKGROUND: PMA induces pulmonary fibrosis in the rabbit (1). Pulmonary fibrosis induced by PMA occurs in the alveolar wall and has the same pattern as idiopathic pulmonary fibrosis (IPF)(2), so this system can be used as an animal model for IPF. PMA also increases the content of alpha-1-antichymotrypsin (ACT) in cultured alveolar macrophages of bronchoalveolar lavages (BAL), and dexamethasone inhibits this PMA-induced increase (3). Here we investigated the role of ACT in pulmonary fibrosis induced by PMA. EXPERIMENTAL DESIGN: Rabbits were treated intratracheally for 6 days with saline, dimethyl sulphoxide (DMSO) used as a solvent of PMA, PMA dissolved in DMSO or PMA plus anti-ACT rabbit serum. BAL samples were obtained. ACT in cell pellet and cell-free fluid of BAL were assayed by radioimmunoassay. Sections of the lung were examined histologically by a point count method. The ratio of fibrosis to elastosis (fibrotic ratio) was evaluated for each rabbit by the ratio of total points of collagen stained by the Azan Mallory method to those of elastic fiber stained by the Elastica van Gieson method. Hydroxyproline (HP) was assayed biochemically, and the amount of HP in the alveolar wall for each rabbit was calculated using the assayed values of HP and the ratio of histologic collagen points in the alveolar wall to those in the lung tissue by a point count method. RESULTS: The fibrotic ratio of the PMA group increased fourfold compared with that of the saline group. The ratio of the PMA plus anti-ACT group decreased and was similar to that of the saline group. The ratio of the DMSO group was about two times as much as that of the saline or the PMA plus anti-ACT groups. The calculated amount of hydroxyproline in the alveolar wall of the PMA group increased and was approximately 1.5-fold compared with that of the saline group. The amount of HP of the PMA plus anti-ACT group decreased and was similar to that of the saline group. In the BAL, the amount and the percentage of ACT in cell pellet per macrophage of the PMA group increased more than those of the saline and DMSO groups. The amount and percentage of the PMA plus anti-ACT group were significantly less than those of the PMA group. Those of the DMSO group were similar to those of the saline group. CONCLUSIONS: These findings suggest that anti-ACT has a suppressive effect on pulmonary fibrosis induced by PMA and that ACT is important in the PMA model of pulmonary fibrosis. PMID- 7474927 TI - Distribution and ontogenesis of tenascin in normal and cystic human fetal kidneys. AB - BACKGROUND: Tenascin is a mesenchymal extracellular matrix glycoprotein transiently expressed during development, mainly at the site of epithelial mesenchymal interactions. It is thought to play a key role in morphogenesis. Little is known about the distribution of tenascin in normal human fetal kidney, and, so far, no data have been reported concerning the distribution of the protein in fetal cystic kidneys. EXPERIMENTAL DESIGN: Using specific mAb and the immunofluorescence technique, we analyzed the distribution of tenascin in normal human embryonic (n = 3), fetal (n = 15), and mature kidneys (n = 4) and in fetuses affected with autosomal recessive polycystic disease (n = 3), autosomal dominant polycystic disease (n = 3), and cystic dysplasia (n = 3). We compared the distribution of this protein with that of fibronectin and types I, III, V, and VI collagens. RESULTS: In normal developing kidneys, tenascin is present in the uninduced blastema and in the mesenchyme around differentiating nephrons. It is homogeneously distributed in the inner cortex and in the medulla. During maturation, tenascin expression persists in the medulla but progressively decreases in the cortex. Tenascin is present in the mesangial area from the S shaped body stage. Both types of polycystic diseases are characterized by a marked and diffuse increase in cortical and medullary expression of tenascin as well as types III, V, and VI collagen. In cystic dysplasia, two types of changes were observed: (a) increased tenascin and interstitial collagen expression in the subcapsular strips of condensed mesenchyme; and (b) heterogeneous medullary tenascin distribution with positive labeling of the condensed mesenchyme surrounding cysts and primitive ducts and negative labeling of the loose interstitial mesenchyme, contrasting with the diffuse accumulation of types III, V, and VI collagen. CONCLUSIONS: In the human fetal kidney, tenascin is expressed by blastema cells and disappears when converted to epithelium. In polycystic diseases, an early increase in tenascin and interstitial collagen expression suggests that renal mesenchyme per se may contribute to the progressive alteration of the kidney. In cystic dysplasia, phenotypic changes in metanephric blastema indicate inappropriate commitment of blastema cells into interstitial cells, leading to the definitive arrest of nephrogenesis; the heterogeneity in tenascin medullary expression underlines the heterogeneity in the mesenchymal cell population. PMID- 7474928 TI - The influence of diets and gut microflora on lectin binding patterns of intestinal mucins in rats. AB - BACKGROUND: The mechanisms responsible for the biosynthesis, storage, secretion, or degradation of intestinal mucins are still unclear. Little is known about the carbohydrate composition of mucins in response to changes in the intestinal lumen, so lectin histochemical techniques were used to study the alterations in glycoconjugate synthesis of mucins in rats under different diets and microfloras. EXPERIMENTAL DESIGN: Nine-week-old germ-free and conventional rats were given either a purified diet of finely powdered ingredients, including cellulose as a source of fiber, or a more coarsely ground commercial diet of natural ingredients containing crude fiber of cereal origin. To mimic the human situation more closely, a group of rats born germ-free, inoculated with a suspension of human feces, and fed a purified diet were used as an experimental model. RESULTS: In rats fed a commercial diet, the surface goblet cells in the small intestine were more intensely labeled with N-acetyl-glucosamine and sialic acid-linked D galactose-specific lectins than in rats fed the purified diet. A similar increased staining with a N-acetylgalactosamine-specific lectin was observed in the large intestine of rats fed a commercial diet. The microbial flora modified the crypt-surface glycosylation of fucosyl and sialic acid residues in the large intestine. The human flora specifically altered the goblet cell glycoconjugates in the surface epithelium. CONCLUSIONS: The significant changes in goblet cell glycoconjugates reflect the adaptation of the intestinal mucosa to different diets and microbial populations. An overall reduction in sialic acid-linked D galactose residues in conventional rats and a loss of crypt-to-surface gradient of fucosyl expression in the large intestine of human flora rats are likely to be due to differing strains of glycosidases in the two microflora. PMID- 7474929 TI - Changes in organization and composition of the extracellular matrix underlying cultured endothelial cells exposed to laminar steady shear stress. AB - BACKGROUND: In blood vessels, the extracellular matrix (ECM) underlying the endothelium supports endothelial cell (EC) attachment, spreading, migration, and proliferation. The structure and composition of the ECM may be modulated by hemodynamic shear stress, which may play a role in the pathogenesis of vascular diseases such as atherosclerosis. EXPERIMENTAL DESIGN: In this study, in vitro effects of fluid shear stress on the ECM of EC were investigated. Cultured bovine aortic EC (BAEC) were exposed to a steady laminar shear stress of 30 dyn/cm2 from 3 to 48 hours, using a parallel-plate flow chamber. Parallel control cultures were maintained under static conditions. The organization of fibronectin (Fn), laminin (Ln), collagen type IV (Col IV), and vitronectin (Vn) was analyzed by immunofluorescence microscopy. Changes in the profile of proteins present in the deoxycholate-insoluble ECM fraction of EC were determined using two-dimensional gel electrophoresis, and the levels of Fn, Ln, and Vn were determined by Western blotting. RESULTS: Fn, Ln, and Col IV exhibited both a granular pattern in cell perinuclear areas and a fibrillar pattern localized underneath EC. On exposure of bovine aortic EC to shear stress, Fn fibrils grouped into thicker tracts of fibrils, and there was a tendency for some of these tracks of fibrils to align with the direction of flow. Ln and Col IV also grouped into thicker fibers, which, in contrast to Fn, were randomly oriented. Vn exhibited a diffuse granular pattern, which did not change in response to shear stress. Consistent increases in the levels of four unidentified acidic proteins (mol wt/pI = 52/4.9, 70/4.7, 70/5.5, and 110/4.4) were observed after 3 to 6 hours of exposure to flow. The level of Fn present in the ECM was decreased twofold 12 hours after exposure of the cell monolayer to flow, and then increased after 24 and 48 hours. The level of Ln showed a twofold increase after 24 and 48 hours of flow, whereas the level of Vn was not altered by shear stress. CONCLUSIONS: These changes in organization and composition observed in the ECM of cultured EC may play a significant role in shear stress-induced morphologic alterations in EC and may represent relevant events in the initiation of atherosclerotic lesions by influencing both EC and smooth muscle cell function. PMID- 7474930 TI - In situ amplification of measles virus RNA by the self-sustained sequence replication reaction. AB - BACKGROUND: The self-sustained sequence replication (3SR) reaction is an isothermal method for nucleic acid amplification that has several features that make it an attractive alternative to PCR. We have studied the feasibility of the in situ 3SR reaction in cells using a measles virus-infected cell line as a model. EXPERIMENTAL DESIGN: The study was carried out in four steps. First, using RNA extracted from a measles-infected Vero Green monkey kidney cell line, conditions for the in vitro amplification of a segment of the nucleocapsid portion of the RNA viral genome were optimized for 420- and 119-bp 3SR products, and the results were compared. Second, 3SR was performed on intact infected cells in suspension, and the amount of RNA product was compared with infected cells without 3SR. Then, the 3SR reaction was conducted on cytospin preparation slides, followed by in situ hybridization for detection of the amplification product. Finally, 3SR was carried out on sections of formalin-fixed, paraffin-fixed, paraffin-embedded cells, and the degree of amplification as detected by ISH was quantified and compared between infected cells with and without 3SR reaction. RESULTS: Specific amplification of measles was observed in each of these types of preparations with an 8.5-fold rate of amplification in paraffin sections of formalin-fixed cells (a mean of 272.5 +/- 65.3 grains/cell after 3SR amplification in comparison to 31.97 +/- 4.2 grains/cell without amplification). CONCLUSIONS: A significant amount of amplification of RNA is possible with in situ 3SR (IS-3SR) and, in combination with ISH, offers several advantages compared with in situ PCR (IS-PCR), such as ease of use, lack of conditions that lead to cell damage, and a specificity for RNA amplification. This is the first report of specific amplification of RNA within cells using the IS-3SR procedure, a technique that has a wide range of potential applications in pathology and molecular biology. PMID- 7474932 TI - Germ cell tumors: unraveling a biological paradox. PMID- 7474931 TI - Enhancement of mRNA in situ hybridization signal by microwave heating. AB - BACKGROUND: Optimization of in situ hybridization protocols is of real interest when trying to detect small amounts of mRNA or when using low concentrations of probes. To enhance the hybridization signal, we have developed a modification of an in situ hybridization (ISH) protocol with radiolabeled cRNA probes. The detailed protocol of ISH used for paraffin sections is also described. EXPERIMENTAL DESIGN: In microwave (MW) heating, the tissue sections are heated in a sodium citrate buffer (0.01 M, pH 6). The effects of the pretreatment with MW were studied on kidneys and adrenals of young rats and on human pathologic samples using [35S]-RNA probes complementary to the mRNAs of some components of the renin angiotensin system. RESULTS: The heating pretreatment with the MW permitted us to obtain an enhancement of the hybridization signal, especially when using low doses of radioactive probes. This enhancement could be evaluated to 60 to 120% by computer-assisted quantification of the signal. Furthermore, the histologic structures and the staining with toluidine blue were not impaired by the heating treatment. CONCLUSIONS: The enhancing effect of the hybridization signal obtained using MW allows shorter autoradiographic exposure times and/or the use of lower concentrations of radioactive probes for the detection of mRNA or the detection of mRNA expressed at the threshold of detection with usual protocols. PMID- 7474933 TI - Pathogenesis of nonimmune glomerulosclerosis: studies in animals and potential applications to humans. PMID- 7474934 TI - Molecular genetic changes in human male germ cell tumors. AB - BACKGROUND: An isochromosome for the short arm of chromosome 12, i(12p), is the most common and characteristic cytogenetic aberration in testicular germ cell tumors. Little is known about the molecular genetic abnormalities of these neoplasms. EXPERIMENTAL DESIGN: A total of 32 loci were studied in DNA from 31 primary testicular germ cell tumors and compared with corresponding normal DNA. The loci map to 17 different chromosome arms, including seven that contain known tumor suppressor genes. Southern blot analysis and PCR-based methods were used. Several microsatellite loci were included to investigate instability (seen as new alleles) at repeat loci. The TP53 tumor suppressor gene was analyzed for point mutations by constant denaturant gel electrophoresis and for expression by immunohistochemistry. Histologic sections of the tumor biopsies were evaluated with regard to components and percentage of intact tumor cells. The growth fraction, representing one component of proliferative activity of the tumor, was assessed by the Ki-67 index. RESULTS: Changes were found at all chromosome arms investigated but at very different frequencies, 5-56% of all tumors. The most frequently affected chromosome arms, those showing loss of heterozygosity or allelic imbalance in more than 40% of the tumors, were 2q, 3p, 3q, 11p, 12p, 18q, and 22q. Complete loss of one allele was often seen at 3p and 11p loci, whereas allelic imbalances dominated on the 2p, 3q, 12p, 18q, and 22q loci tested. No mutations were detected within four known mutational hot spots of TP53, but positive immunostaining with two TP53 Ab was seen in 9 of 14 tumors. Most tumors (26 of 31) showed positive immunostaining with Ki-67. Microsatellite instability was not observed. CONCLUSIONS: High frequencies of loss of heterozygosity and allelic imbalance at several loci indicate that inactivation of several tumor suppressor genes may be of importance in developing testicular germ cell tumors. The increase in copynumber of 12p alleles seen in several tumors is likely to reflect one or more 12p isochromosomes. Our findings do not indicate that TP53 plays any major pathogenic role in this tumor type, nor was there any indication that defect repair genes, causing microsatellite instability in other cancers, participate in the progression of testicular cancer. PMID- 7474936 TI - Transforming growth factor-beta 1: comparative immunohistochemical localization in human primary and metastatic prostate cancer. AB - BACKGROUND: We have shown previously that primary prostate cancer demonstrates significant extracellular accumulation of transforming growth factor-beta 1 (TGF beta 1). To further investigate the potential role of TGF-beta 1 in prostate cancer progression, we evaluated an expanded series of primary prostatic carcinomas and associated lymph node metastases. EXPERIMENTAL DESIGN: Prostate tissue samples from 37 patients were examined. Three were organ donors, all less than 30 years of age, whose prostates were included as normal controls. Eighteen had undergone radical prostatectomy and pelvic lymph node dissection. Eleven were without evidence of metastasis, whereas seven were found to have prostate cancer within at least one of their pelvic lymph nodes. Sixteen had undergone transurethral resection of the prostate for benign disease, yet all had prostate cancer within the resected specimen (15 were stage T1b; 1 was stage T1a). Twelve of the patients with stage T1b disease underwent pelvic lymph node dissection and implantation of radioactive gold seeds. All 12 had prostate cancer in at least one lymph node. All specimens were examined for the level of expression and localization of TGF-beta 1 by immunohistochemistry using Ab that distinguish intracellular from extracellular TGF-beta 1. RESULTS: Normal prostate tissue and benign prostatic hyperplasia demonstrated negative or weak intracellular and extracellular staining for TGF-beta 1. By comparison, 29 of 34 primary prostate cancers showed extensive extracellular TGF-beta 1 staining with pronounced intracellular accumulation within epithelial cells in 13 of 34 patients. There was no difference in the staining pattern for extracellular TGF-beta 1 between primary cancers with and without pelvic lymph node metastases. However, in primary cancers without pelvic lymph node metastases, only 1 of 15 patients showed strong intracellular staining for TGF-beta 1 compared with 12 of 19 primary tumors with metastatic disease. In addition, only 2 of 19 lymph node metastases demonstrated weak extracellular TGF-beta 1 staining, but all 19 contained intracellular TGF-beta 1. CONCLUSIONS: These findings confirm our previous observation that prostate cancer exhibits enhanced intracellular and extracellular accumulation of TGF-beta 1 relative to normal prostate tissue and benign prostatic hyperplasia. In addition, our study documented a significantly more pronounced accumulation of intracellular TGF-beta 1 in primary prostate cancer with metastasis than in primary tumors without metastasis. Moreover, although the pattern of intracellular TGF-beta 1 staining observed in the primary tumor is maintained in the metastasis, a lack of extracellular accumulation of TGF-beta 1 in the metastatic site was noted. This differential pattern may be biologically important and could conceivably reflect a role for TGF-beta 1 in disease progression. PMID- 7474935 TI - Tissue and cellular distribution of an adhesion molecule in the carcinoembryonic antigen family that serves as a receptor for mouse hepatitis virus. AB - BACKGROUND: The receptor for the murine coronavirus mouse hepatitis virus (MHV) A59, called MHVR or Bgp1a, is a glycoprotein in the carcinoembryonic antigen family of the Ig superfamily. Biliary glycoprotein (Bgp) isoforms play a role in cell adhesion, bile acid transport, and ecto-ATPase activity. MHV-resistant SJL/J mice express a different allele of Bgp1 called Bgp1b. Analysis of the tissue and cellular distribution of Bgp1 proteins can therefore provide new insight on both cellular functions and MHV-A59--induced pathogenesis. EXPERIMENTAL DESIGN: Bgp1 expression was analyzed in the digestive, respiratory, endocrine, urinary, and central nervous systems of adult BALB/c and SJL/J mice by immunocytochemistry and immunoelectron microscopy using a monoclonal Ab specific for the N-terminal domain of the Bgp1a proteins and polyclonal rabbit anti-Bgp1, which recognizes both the Bgp1a and Bgp1b proteins. The function of Bgp1 proteins as viral receptors was tested on tissue sections by a virus binding assay. MHV-A59 replication was analyzed by immunocytochemistry. RESULTS: Bgp1 expression was observed on membranes of epithelial cells (including hepatocytes, intestinal, endocrine, and respiratory epithelial cells), kidney proximal tubules, and endothelial cells in many tissues. It was usually localized at the apical pole of the cells and, when present, on the brush borders and the cilia. A new direct virus binding assay showed that MHV attachment onto cells correlates with Bgp1 expression. Viral infection was detected in hepatocytes, lymphoid tissue, and the exocrine pancreas but not in endocrine cells, enterocytes, kidney, or respiratory cells. In the central nervous system, no immunolabeling of neurons or glial cells was found with anti-Bgp1 Ab. CONCLUSIONS: Bgp1 proteins are present on BALB/c and SJL/J epithelia and endothelia. These glycoproteins might be involved in cell-to cell contacts, for example between hepatocytes. On BALB/c mice, Bgp1a expression is consistent with the tropism of MHV-A59 for the liver. However, Bgp1a was also expressed on cells that were not infected by MHV-A59. PMID- 7474937 TI - Therapy of experimental human brain tumors using a neuroattenuated herpes simplex virus mutant. AB - BACKGROUND: Engineered herpes simplex virus (HSV) strains previously have been shown to offer a potential therapeutic alternative to conventional treatment modalities for brain tumors. Because HSV Type 1 strain 1716 has a deletion in the gamma 34.5 neurovirulence gene that renders it avirulent in the mouse central nervous system, we have assessed its potential to induce selective lysis of tumor cells versus neurons in vitro and in vivo. EXPERIMENTAL DESIGN: To do this, we studied parental HSV-1 strain 17+ and strain 1716 using human embryonal carcinoma cells (NT2 cells). These cells resemble neuronal progenitor cells and can be induced to differentiate into neurons (NT2N) with retinoic acid. Intracerebral grafts of NT2 cells into the brains of nude mice resulted in lethal brain tumors, and grafts of NT2N cells resulted in the integration of NT2N cells. RESULTS: In vitro studies showed that strain 1716 replicates in and spreads on monolayers of NT2 cells but not in NT2N cells. In vivo, strain 1716 replicated preferentially in NT2 tumors as evidenced by immunohistochemical staining for viral antigens, by in situ hybridization for HSV-specific transcripts, and by titration of virus from brains with tumor after intracranial injection of the virus into these mice. The temporal regression of NT2 tumors in mice treated with strain 1716 was demonstrated in vivo by magnetic resonance imaging. Electron microscopy and studies of DNA fragmentation suggested that regression of NT2 brain tumors in strain 1716-treated mice was mainly caused by a nonapoptotic, lytic mode of cell death. Finally, strain 1716-treated NT2 tumor-bearing mice survived more than twice as long as mock-treated tumor-bearing mice, and these differences in survival (25 vs. 9 weeks) were statistically significant (p < 0.03). CONCLUSIONS: We conclude from these studies that strain 1716 induces regression of human neural tumors established in the brains of nude mice, resulting in their prolonged survival. PMID- 7474939 TI - Rat aortic smooth muscle cells become pericytes during angiogenesis in vitro. AB - BACKGROUND: We previously reported that the intimal endothelium of the rat aorta switches to a microvascular phenotype during angiogenesis in vitro. The microvessels formed by the rat aortic endothelium are coated with pericytes. The purpose of this study was to evaluate the relation of the pericytes to the angiogenic process and to identify the site of origin of these cells in the aortic wall. EXPERIMENTAL DESIGN: Rings of rat aorta were cultured in collagen gel under serum-free conditions. The formation of a pericyte coating around aorta derived microvessels was evaluated by counting pericytes and microvessels in the living cultures. Pericytes and endothelial cells were studied by immunohistochemistry, lectin labeling, electron microscopy, 3H-thymidine labeling followed by autoradiography, and time-lapse video microscopy. The capacity of aortic smooth muscle cells to differentiate into pericytes was studied by coculturing intimal- or medial-derived smooth muscle cells with endothelial cells in a collagen gel overlay assay that induced reorganization of endothelial cells into microvessels. RESULTS: Microvessels during the early stages of angiogenesis were composed primarily of endothelial cells. As vascular proliferation decreased, the microvessels became coated with pericytes. The pericytes migrated from the root to the tip of the microvessels using the endothelium as a surface for attachment, proliferation, and contact guidance. The pericytes were continuous with the myointimal endothelial cells of the cultured aorta. Pericytes and myointimal cells were positive for alpha-smooth muscle actin and vimentin and were actively engaged in DNA synthesis. Treatment of the cultures with heparin caused a marked reduction in the number of pericytes. Smooth muscle cells isolated from the intimal aspect of the rat aorta migrated toward the endothelium and differentiated into pericytes when cocultured with microvessels formed by isolated endothelial cells in a collagen gel overlay assay. Conversely, smooth muscle cells isolated from the deep layers of the media had no significant endothelial tropism and failed to differentiate into pericytes. CONCLUSIONS: This study demonstrates that the rat aorta contains a subpopulation of intimal/subintimal smooth muscle cells that differentiate into pericytes during angiogenesis in vitro. These cells have a distinct endothelial tropism and respond to endothelial cues by contributing to the differentiation and maturation of microvessels. Smooth muscle cells of rat aortic intimal/subintimal origin can be used as a source of pericytes for the in vitro assembly of histotypic microvessels. PMID- 7474938 TI - Expression of the basement membrane heparan sulfate proteoglycan (perlecan) in human synovium and in cultured human synovial cells. AB - BACKGROUND: Perlecan is a heparan sulfate proteoglycan that has a core protein of 466 kDa and is composed of five modules, four of which share a high degree of homology with a variety of biologically important molecules including the low density lipoprotein receptor, laminin, and the neural cell adhesion molecule. Previously, this specialized proteoglycan, characteristic of the basement membrane, was not identified as a constitutive extracellular component of human synovium. EXPERIMENTAL DESIGN: Using human synovium and cultured human synovial cells, we examined perlecan gene expression in human synovium, a tissue lacking a classic basement membrane. The modulation of gene expression of the perlecan core protein by transforming growth factor-beta (TGF-beta) and basic fibroblast growth factor (FGF) was examined in cultures of normal human synovial cells, and by metabolic labeling using radioactive sodium sulfate, the question of whether this specialized protein is produced by synovial cells as a proteoglycan was addressed. RESULTS: Cultures of normal human synovial cells were shown to contain the large 14.5 kb perlecan mRNA and produced substantial amounts of perlecan core protein as shown by immunohistochemistry employing specific human perlecan Ab. Immunohistochemical detection showed intense staining in the intimal and subintimal layers of human synovial membrane. Moreover, the perlecan core protein was shown to be up-regulated by TGF-beta and down-regulated by basic FGF. In addition, a sizable portion (approximately 25%) of the synovial cell-produced proteoglycan was shown to contain heparan sulfate, providing evidence that synovial cell perlecan is produced as a proteoglycan. CONCLUSIONS: The presence of perlecan in human synovium and the modulation of the biosynthesis of its core protein by TGF-beta and FGF suggest that, in addition to its structural role, this unique heparan sulfate proteoglycan may be involved in normal synovial membrane function and in the pathogenesis of arthritis. PMID- 7474940 TI - The effects of high glucose on human endothelial cell growth and gene expression are not mediated by transforming growth factor-beta. AB - BACKGROUND: Because accumulation of extracellular matrix is a prominent characteristic of the microangiopathy that complicates long-term diabetes, a pathogenetic role for transforming growth factor beta (TGF-beta) is being considered. Having observed that glucose levels mimicking diabetic hyperglycemia induce in vitro endothelial cell overexpression of extracellular matrix molecules, decreased replication, and increased levels of TGF-beta mRNA, we have examined whether the effects of high glucose are mediated by autocrine TGF-beta. EXPERIMENTAL DESIGN: TGF-beta levels were measured by bioassay in the media conditioned by human umbilical vein endothelial cells cultured in the presence of high (30 mM) or normal (5 mM) glucose concentrations. The effect of high glucose was tested on the proliferation of two epithelial cell lines, one (Mv1Lu) exquisitely sensitive to TGF-beta and the other (DR mutants) insensitive to the cytokine. To examine whether high glucose and TGF-beta affect cellular programs in a similar manner, the effects of high glucose and exogenous TGF-beta were compared on proliferation and gene expression of endothelial cells. RESULTS: Media conditioned by endothelial cells cultured in high or normal glucose contained similar amounts of TGF-beta (4.9 +/- 3.5 and 3.7 +/- 2.5 ng/10(6) cells, respectively (mean +/- SD)), all in the latent form. The replication of parental Mv1Lu cells and their DR mutants was decreased by high glucose to the same extent. Whereas the inhibitory effect of high glucose on endothelial cell replication was reversible, that of TGF-beta was not. Both perturbations induced up-regulation of fibronectin expression, but the effects were additive. Only TGF beta induced overexpression of Type IV collagenase. CONCLUSIONS: These combined observations indicate that (a) endothelial cells exposed to high glucose do not secrete TGF-beta in excess of control cells, (b) there are growth-inhibitory effects of high glucose that are independent of TGF-beta, and (c) high glucose and TGF-beta exert their effects through distinct pathways and at different loci. PMID- 7474941 TI - Immunogold localization of high-affinity glucose transporter isoforms in normal rat kidney. AB - BACKGROUND: Facilitative glucose transporters (GLUT) have unique kinetic characteristics and distributions suited to the functions of the tissues in which they reside. However, little is known about their individual roles in renal glucose metabolism, and previous investigations of renal GLUT expression have been extensive only with respect to their mRNA levels. We provide here a complete analysis of three GLUT isoforms along the nephron using a sensitive immunodetection method. EXPERIMENTAL DESIGN: Normal rat kidneys were harvested, fixed in paraformaldehyde, and embedded in either paraffin or resin as required for immunogold labeling of individual GLUT Isoforms 1, 3, and 4. Samples were evaluated by light microscopy and selected regions analyzed by high resolution optical scanning with computer-assisted detection of immunogold-labeled GLUT at the subcellular level. We describe, compare, and related to the local patterns of glucose metabolism the cellular and subcellular expression patterns of these GLUT along the nephron. RESULTS: GLUT1 was most intensely labeled in the medullary thick ascending limbs of Henle, cortical collecting ducts, and inner medullary collecting ducts. In contrast, GLUT3 was most prominent in the inner medullary collecting ducts and GLUT4 in medullary thick ascending limbs of Henle. All three GLUT were detected in glomerular tufts, and GLUT1 was also detected in parietal epithelial cells. The predominant subcellular distributions in tubule cells were: basolateral and basolateral/cytoplasmic for GLUT1; basolateral and cytoplasmic for GLUT3; and perinuclear/cytoplasmic for GLUT4. GLUT 1 and 3 expressions were confirmed in specific regions by immunoblotting. CONCLUSIONS: 1) GLUT 1, 3, and 4 are expressed in both glomeruli and renal tubules. 2) The unique GLUT expression patterns along the renal tubules suggests unique functional roles for these isoforms. 3) The renal cortex demonstrates lesser labeling intensity for the high affinity GLUT compared with the medulla, where higher rates of glucose oxidation and glycolytic metabolism are paralleled by higher GLUT labeling intensities. PMID- 7474942 TI - CD26 expression in leprosy and other granulomatous diseases correlates with the production of interferon-gamma. AB - BACKGROUND: Leprosy represents a spectrum of clinical manifestations that reflect the immune response to antigens of Mycobacterium leprae. The tuberculoid form of leprosy, which is characterized by an organized development of granulomas, has recently been correlated with a Th1-like immune response. The lepromatous form of leprosy, with a characteristic lack of cellular immunity, has been correlated with a Th2-like immune response to mycobacterial antigens. Dipeptidylpeptidase IV (CD26) is an ectopeptidase that is expressed in various tissues; in the hemopoietic system, it is predominantly expressed by T cells. EXPERIMENTAL DESIGN: We stained frozen sections of skin biopsies obtained from patients with different forms of leprosy, sarcoidosis, and Piringer's lymphadenitis. Sections were stained for interferon-gamma (IFN-gamma) and CD26 with the alkaline phosphatase anti-alkaline phosphatase technique and in two-color stainings by immunofluorescence. RESULTS: We found strong signals for IFN-gamma and for CD26 in all investigated cases of tuberculoid leprosy. In contrast, in all biopsies taken from patients with lepromatous leprosy, we found no or very weak signals for these antigens. By immunofluorescence double-labeling, we could show that IFN gamma and CD26 were expressed by the identical cell population. We confirmed this correlation of CD26 expression and IFN-gamma production in other granulomatous inflammatory reactions such as sarcoidosis and Piringer's lymphadenitis. CONCLUSIONS: From our results, we conclude that a high expression of CD26 may be suggestive of Th1-like immune reactions. PMID- 7474943 TI - Pathology of experimental inhalation anthrax in the rhesus monkey. AB - BACKGROUND: The inhalation form of anthrax, although rare, is nearly always fatal because of the rapid progression of the disease with little host response until the terminal stages of the disease. The Gulf War heightened the concern that anthrax could be used as a biologic weapon. Past studies modeling pathologic changes in human inhalation anthrax have used the rhesus monkey. EXPERIMENTAL DESIGN: We studied pathologic changes in the rhesus monkey model of inhalation anthrax. Gross examination as well as light and electron microscopy were used to define pathologic alterations. Immunolabeling techniques were used to identify the anthrax bacillus by light and electron microscopy. RESULTS: Gross changes included hemorrhage in mesenteric (54%) and tracheobronchial (46%) lymph nodes, meninges (38%), lungs (31%), and small intestinal serosa (31%). Histopathologic changes included suppurative meningitis (77%); hemorrhages in the meninges (54%), neuropil (31%), and pulmonary alveoli (31%); and pneumonia (15%). Spleens and various lymph nodes from all monkeys had one or more of the following changes: hemorrhage, acute inflammation, extracellular bacilli, lymphocytic depletion, and histiocytosis. Spleens of two monkeys were devoid of extracellular bacilli, but degraded intrahistiocytic bacilli reacted with Ab to Bacillus anthracis cell wall polysaccharide. CONCLUSIONS: In our study, compared with previous reports, meningitis and mesenteric lymph node hemorrhages were more common, whereas mediastinal and tracheobronchial lymph node hemorrhages were less common. Immunostaining highlighted intracellular bacilli that would have been otherwise missed by light microscopic examination. PMID- 7474944 TI - Amyloid fibril composition and transthyretin gene structure in senile systemic amyloidosis. AB - BACKGROUND: In many different forms of amyloidosis, transthyretin (TTR) comprises the major amyloid fibril protein. In the familial forms, various TTR mutations are linked to disease. This study was designed to characterize the components of the TTR-derived amyloid fibril protein in senile systemic amyloidosis and to determine whether any mutation in the TTR gene was present. EXPERIMENTAL DESIGN: Heart tissues from two patients with advanced senile systemic amyloidosis were studied. Amyloid fibrils were extracted and the amyloid fibril protein purified. The relationship between full-length and fragmented TTR and the amino acid sequence of the TTR were determined. The TTR gene was studied by single-strand conformation polymorphism analysis or direct sequencing. RESULTS: In both cases, the amyloid deposits contained full-length TTR and a complex mixture of TTR fragments. The fragments, most of which had their N-termini at positions 46-52, predominated. No amino acid substitution was identified. The N-terminal fragment (1-45) was not identified in either patient. In each case, the four exons of the TTR gene were of normal sequence. CONCLUSIONS: In familial amyloidosis resulting from deposition of TTR Met 30 (Swedish-type familial amyloidosis), full-length TTR molecules (some mutant) usually predominate, and TTR fragments lacking three of the eight beta-strands (nonmutant) form a major part of the fibril in senile systemic amyloidosis. This may indicate a difference in the fibrillogenesis between these two forms of TTR-derived amyloidosis. We propose that the name senile systemic amyloidosis be used only for normal-sequence TTR amyloidosis occurring in advanced age. PMID- 7474945 TI - Relaxin depresses platelet aggregation: in vitro studies on isolated human and rabbit platelets. AB - BACKGROUND: Relaxin, a peptide hormone of ovarian origin, has been shown to cause a striking dilatory action on microvessels in different organs. In our recent studies, relaxin has been shown to stimulate the production of nitric oxide, a powerful vasodilatory agent, in several targets. Nitric oxide also inhibits platelet aggregation. This prompted us to search for a role of relaxin in platelet function. EXPERIMENTAL DESIGN: The effect of relaxin on platelet aggregation was studied in isolated human and rabbit platelets. The samples were incubated with relaxin at different concentrations and then stimulated with collagen or thrombin. Aggregation and intracellular levels of cGMP and Ca2+ were determined. In some experiments, inhibitors or potentiators of nitric oxide activity were also used to clarify whether the mechanism of action of relaxin involves the L-arginine-nitric-oxide pathway. Electron microscopy of platelets treated and not treated with relaxin was also carried out. RESULTS: Preincubation of the platelets with relaxin before stimulation with proaggregants resulted in a significant, concentration-dependent inhibition of platelet aggregation, accompanied by an elevation of intraplatelet cGMP and a decrease in the rise of cytosolic Ca2+ levels. The effect of relaxin appeared to be mediated through nitric oxide. Ultrastructurally, relaxin was shown to hinder the conformational changes and granule exocytosis usually occurring in platelets during aggregation. CONCLUSIONS: This newly recognized antiaggregatory property of relaxin, together with the vasodilatory and hypotensive activities of the peptide demonstrated in previous studies, allows for this hormone to be regarded as a protective agent against thrombotic and hypertensive disorders of pregnancy and cardiovascular diseases. PMID- 7474946 TI - Induction of EGF-receptor tyrosine kinase during early reparative phase of gastric mucosa and effects of aging. AB - BACKGROUND: Although the gastric mucosa of adult healthy animals possesses a remarkable capacity to promptly repair its mucosal architecture after an acute injury, aging attenuates this process. We hypothesize that certain tyrosine kinases (Tyr-k), specifically the enzyme associated with EGF-receptor (EGF-R), may play a role in this process. The present investigation was undertaken to evaluate the role of this enzyme in the early reparative phase of the gastric mucosa in young and aged rats. EXPERIMENTAL DESIGN: In our initial effort to test the hypothesis, we examined the changes in both total and EGF-R-associated Tyr-k activities in the gastric mucosa of young adult rats (4-months old) during the first 60 minutes after hypertonic saline (2 M NaCl; 1.5 ml/130 g body weight) induced injury. Because the maximal stimulation (90-100% over the controls) in both total and EGF-R-associated Tyr-k occurred at 30 minutes after injury, we used this time point to perform the next experiment, in which groups of young and aged rats were given (intragastically) 2 M NaCl or water. One of the young and aged groups of rats was also injected (i.p.) with the Tyr-k inhibitor tyrphostin 51 (300 micrograms/kg body weight) 60 minutes before injury. The gastric mucosa was assayed for EGF-R Tyr-k activity and tyrosine phosphorylation and expression of EGF-R, phospholipase C (PLC) activity and relative concentration and tyrosine phosphorylation of PLC-gamma 1, as well as transforming growth factor-alpha (TGF alpha) levels. RESULTS: Basal EGF-R Tyr-k activity and the extent of tyrosine phosphorylation of EGF-R, as well as PLC activity, were all found to be higher in the gastric mucosa of aged than in young rats. Although 30 minutes after injury, EGF-R Tyr-k activity, tyrosine phosphorylation of EGF-R, and relative abundance of the receptor were all increased in the gastric mucosa of both young and aged rats, the magnitude of stimulation of each of the parameters was found to be considerably lower in aged than in young rats, compared with the corresponding basal levels. A similar phenomenon was also observed for PLC activity and tyrosine phosphorylation of PLC-gamma 1. The relative concentration of mucosal PLC-gamma 1 level was, however, not affected by injury in either young or aged rats. Tyrphostin greatly attenuated the injury-induced increases in the above mentioned parameters in both young and aged rats. In young but not in aged rats, injury caused a significant increase in mucosal TGF-alpha levels. CONCLUSIONS: We conclude that (a) activation of EGF-R Tyr-k is an important event in the early reparative process of the gastric mucosa, and (b) local production of TGF-alpha may play an important role in regulating the activation of EGF-R Tyr-k. PMID- 7474947 TI - In situ detection of cyclosporin A: evidence for nuclear localization of cyclosporine and cyclophilins. AB - BACKGROUND: The immunosuppressant cyclosporin A (CsA) forms a trimolecular complex with cyclophilin (CPH) and calcineurins (CN) and inhibits CN phosphatase activity. Inhibition of CN phosphatase by CsA prevents the dephosphorylation of a nuclear factor in the cytosol and its nuclear translocation to the nucleus. EXPERIMENTAL DESIGN: The intracellular distribution of CPH and CN was investigated in permeabilized Jurkat T lymphocytes and MRC fibroblasts using biochemical techniques and confocal microscopy. The site of CsA binding was identified in situ using a photoaffinity label derivative of CsA followed by immunodetection. RESULTS: Cyclophilin A (CPH-A) and CN display essentially a cytosolic localization by immunofluorescence, and additional nuclear CPH-A and CN are evidenced by Western blot analysis of purified nuclei and immunofluorescence. By contrast, cyclophilin B (CPH-B) has a punctuate and reticular distribution pattern in cytoplasm, indicating an association with the endoplasmatic reticulum, but its main location is in the nuclear matrix, sparing the nucleolar region. For the in situ detection of CsA binding sites, a photolabile cyclosporine derivative (PL-CS) was used that allowed the detection of covalently bound CsA by Ab. Using the biologically active PL-CS, a punctate cytoplasmatic and nuclear immunoreactivity was obtained, which was specific and competed only with active cyclosporine derivatives. Nuclear CPH-A and -B were labeled by PL-CS, and trimolecular complexes of labeled CPH and CN were obtained by chemically cross linking nuclear extracts. CONCLUSIONS: We describe herein the accessibility of CsA to the nucleus, the presence and labeling in situ of nuclear CPH and CN. The current models of CsA action predict that CsA-CPH complexes inhibit CN activity in the cytosol. However, our present findings invite the hypothesis that CPH may capture the drug into the nucleus and target regulatory proteins or transcriptional control elements. PMID- 7474948 TI - Automatic vessel segmentation and quantification of the rat aortic ring assay of angiogenesis. AB - BACKGROUND: Pharmacologic control of angiogenesis is a promising new approach to the treatment of a variety of pathologic conditions including cancer. The recently developed in vitro rat aortic ring model provides a simple, reproducible assay for discovering angiogenic agonists and antagonists. However, quantification of results in this assay is time consuming, tedious, and subjective, because it involves visual inspection of images and manually counting the newly formed microvessels extending from the cultured aortic ring. This report describes an automated image analysis-based procedure for quantification of this assay that overcomes these difficulties. EXPERIMENTAL DESIGN: The designed image processing algorithm segments the vessels from gray scale images. A high-pass filter is used, and the results are separated into nonvascular and vessel compartments based on object size and shape. Quantification relies on identification of vessels intersecting a closed transect set a fixed distance from the aortic ring. The number and the total area of these vessels are determined. The entire operation has been automated and packaged in an application called Vessels. RESULTS: The correlation between computer-determined vessel area/vessel number and visual microvessel count is high (r2 = 0.91 and r2 = 0.86, respectively). CONCLUSION: Vessels offers high-speed, fully automatic batch processing including production of a hard copy for documentation. The application runs on the Apple family of computers. On a Quadra 800, the application can process approximately 30 images/hour, which is approximately 2.5 times faster than manual quantification of this assay. PMID- 7474949 TI - How's your domestic violence I.Q.? PMID- 7474950 TI - A call for physicians to help end the cycle of domestic violence. PMID- 7474951 TI - Protecting women and children from physical, sexual, and psychological abuse. AB - Domestic violence is a serious public health problem. The "epidemic" is becoming too obvious to hide, too widespread to deny, and too much of a health hazard to ignore. The victims are children, women, the elderly, and the society as a whole. Family members, government, educational and penal agencies, civic and medical societies, physicians, and citizens at large are all involved in domestic violence prevention. By having a high index of suspicion, physicians are in an ideal position to prevent, assess, identify, and treat victims of domestic violence and its associated problems. PMID- 7474952 TI - Responding to child maltreatment. PMID- 7474953 TI - Elder abuse. PMID- 7474955 TI - Breaking the cycle of violence. PMID- 7474954 TI - Domestic violence: impact on psychiatric medicine. AB - The 1984 Attorney General's Task Force Report on Domestic Violence attested: Anyone who lives in a violent home experiences an essential loss. The one place on earth where they should feel safe and secure has become a place of danger...the shadow of domestic violence has fallen across their lives and they are forever changed. One report states that the victims of the crime of domestic violence include "not only the people who die from injuries, but the family members who daily endure the psychological, emotional and spiritual abuse and pass on the emotional scars and violent behavior to one generation after another." If we are to stop this cycle of violence, we as physicians must step forward and assume our personal and professional responsibilities. PMID- 7474956 TI - Let's focus on prevention, not treatment, of child abuse and neglect. PMID- 7474957 TI - The clinical studies trust fund--your charity. PMID- 7474958 TI - What is your diagnosis? Multiple osteolytic myeloma with fracture. PMID- 7474959 TI - Intestinal permeability testing in dogs with diet-responsive intestinal disease. AB - Fifteen dogs with signs of small and, or, large bowel disease that responded clinically to an exclusion diet were studied, using differential sugar absorption as an objective parameter of the mucosal response to the diet. Intestinal permeability and function were assessed by determining the urinary excretion ratios of lactulose/rhamnose and xylose/3-O-methylglucose, respectively, following oral administration of a mixture of these four sugars. Five dogs, all retrievers, were tentatively diagnosed as having dietary hypersensitivity, based upon resolution of clinical signs and normalisation of high intestinal permeability following an exclusion diet and recurrence of signs (in four of five dogs) upon challenge with the original diet. The fifth dog did not become symptomatic when challenged, but intestinal permeability increased. The remaining 10 dogs were diagnosed as having food intolerance, based upon clinical improvement on an exclusion diet, relapse on challenge with their original diet, but lack of improvement in intestinal permeability. These findings suggest that a differential sugar absorption test may be useful to determine the reasons for clinical response to exclusion diets. Demonstration of increased intestinal permeability with subsequent normalisation following an exclusion diet may be useful in the diagnosis of dietary hypersensitivity, while persistent abnormalities in intestinal permeability are suggestive of underlying intestinal disease and food intolerance. PMID- 7474960 TI - Measurement of skin hydration in normal dogs and in dogs with atopy or a scaling dermatosis. AB - The role of the stratum corneum both as a barrier to water loss from the body and as a biological microenvironment is well recognised, although most discussion of these topics is expressed in qualitative terms. This paper describes the use of techniques for the measurement of some aspects of the skin microclimate, and their application to canine skin. The techniques used were the assessment of skin moisture, hygroscopicity and water-holding capacity by measurement of skin electrical capacitance. Meaningful deductions regarding the water dynamics of certain skin sites can be drawn, and it was shown that significant differences in hydration exist between different skin sites. The techniques were also used to show that scaly skin is 'drier' than normal skin, and that the water dynamics of the non-lesional skin of atopic dogs does not differ from that of normal dogs. PMID- 7474961 TI - Incidence of the gene mutation causal for rod-cone dysplasia type 1 in Irish setters in the UK. AB - A survey to establish the UK prevalence of the gene mutation causing the rod-cone dysplasia type one (rcd1) form of generalised progressive retinal atrophy (gPRA) in Irish setters was carried out. The dogs were selected by members of two Irish setter breed societies to provide examples from most of the main breeding lines in the UK. A total of 210 Irish setters were tested and one bitch was found to be a carrier of the rcd1 mutation. These results show that although a confirmed case of rcd1 has not been reported in Irish setters in the UK for over a decade the gene is still present in the gene pool. PMID- 7474962 TI - Antebrachial deformities in the dog: treatment with external fixation. AB - The results of 12 dogs with antebrachial deformity treated by ulnar ostectomy, radial osteotomy and external skeletal fixation are presented. Postoperative complications were seen in only one dog; a delayed union requiring placement of autogenous cancellous bone graft at a second surgery to achieve healing. Postoperatively, owners reported limb function was good in almost all cases, although they were less pleased with the cosmetic appearance. PMID- 7474963 TI - Vaginal cysts causing tenesmus in a bitch. AB - An eight-year-old female border collie was referred with a two-month history of faecal tenesmus. Ultrasonography and radiology identified fluid-filled masses in the cranial vagina. The fluid was drained from the vaginal cysts via an episiotomy and the clinical signs resolved. Histological examination confirmed that the cysts were of urogenital origin. This is believed to be the first report of vaginal cysts causing faecal tenesmus in the dog. PMID- 7474964 TI - Giant cell tumour of bone in a cat. AB - A domestic shorthaired cat had a firm, non-painful swelling of the left antebrachium. Radiographs showed an expansile, multiloculated lesion in the radius. The mass gradually enlarged over the following four and a half months and the animal became completely lame. The histological diagnosis was giant cell tumour of bone; this is a rare neoplasm of bone in veterinary patients. Giant cell tumour of bone should be considered in the differential diagnosis of bone lesions having an expansile, multiloculated radiographic appearance. PMID- 7474965 TI - Use of a pedicled rectus abdominis muscle flap for repair of a failed vesicourethral anastomosis in a dog. AB - A 10-year-old mixed breed dog underwent a prostatectomy for an adenocarcinoma. Within three days the dog had signs of a uroperitoneum. Re-exploration revealed that a portion of the vesicourethral anastomosis had failed. Primary suturing of the tissue along the site of dehiscence following debridement of the devitalised tissue was impossible. A rectus abdominis muscle flap was created and successfully used to repair the site of leakage. PMID- 7474966 TI - Acute intrinsic renal failure and blood coagulation disorders after a snakebite in a dog. AB - Acute intrinsic renal failure was diagnosed in a two-year-old, male, German shepherd dog following a Vipera aspis bite. Clinical signs included depression, hypersalivation, vomiting, tachypnoea, abdominal pain, splenomegaly, oliguria with haematuria and haemolysed serum. Leucocytosis with a shift to the left, thrombocytopenia, prolonged coagulation times (activated partial thromboplastin time, prothrombin time and thrombin time), hypofibrinogenaemia, azotaemia and hyposthenuria were the most prominent laboratory abnormalities. Histopathological evaluation of the kidneys showed a discrete glomerular hypercellularity, mesangial lysis and renal tubules filled with many hyaline casts and some necrotic cells. PMID- 7474967 TI - Rabies in a changing world. PMID- 7474968 TI - Part-word repetitions by persons who stutter: fragment types and their articulatory processes. AB - One of the core features of developmental stuttering is part-word repetition followed by fluent production (resolution) of the target word. The purpose of the study is to compare the spectro-temporal dimensions of fragments with the spectro temporal dimensions of the resolution to define fragment types and develop articulatory interpretation of termination-restart cycles. From the acoustic recordings of six adult males who stutter, 142 stuttering events on words beginning with stops were isolated, excerpted, and digitized. The stuttering events occurred during either spontaneous speech or reading. The consecutive articulatory phases of fragments were classified and compared with the corresponding phases in resolutions. We found that (a) there are two basic types of fragments--those with vowels and those without, (b) the type of fragment produced is strongly influenced by the voicing status of the stop, (c) the fragments without vowels tend to have longer stop closure duration than the stops in the resolutions, (d) there are two subtypes in fragments with vowels--those with vowels shorter than and those with vowels longer than the vowels in the resolutions, (e) the shorter vowels differ spectro-temporally from the vowels in resolutions, and (f) the longer vowels differ temporally from the vowels in the resolutions. We discuss the articulatory implications of the acoustic data for each fragment type in the context of speech as an act designed to achieve contextually conditioned acoustic goals. PMID- 7474969 TI - Phonatory and articulatory changes associated with increased vocal intensity in Parkinson disease: a case study. AB - This study examined changes in voice and speech production in a patient with Parkinson disease as he increased vocal intensity following 1 month of intensive voice treatment. Phonatory function and articulatory acoustic measures were made before and after treatment as well as 6 and 12 months later. Pre- to post treatment increases were documented in sound pressure level in sustained phonation, syllable repetition, reading, and monologue. Consistent with mechanisms of intensity change reported in normal speakers, corresponding improvements were measured in estimated subglottal pressure, maximum flow declination rate, laryngeal airway resistance, open quotient, EGGW-25, harmonic spectral slope, and maximum vowel duration. Measures of phonatory stability in sustained phonation and semitone standard deviation in reading and speaking showed changes accompanying increased vocal intensity. In addition, changes were measured in articulatory acoustic parameters (vowel and whole word duration, transition duration, extent and rate, and frication duration and rise time) in single-word productions. These findings indicate that this patient increased his vocal intensity using phonatory mechanisms that have been associated with the nondisordered larynx. In addition, the increased vocal intensity led to changes in articulation that were not targeted in treatment. PMID- 7474970 TI - Pathologic voice type and the acoustic prediction of severity. AB - We hypothesized that acoustic measures would predict dysphonic severity with differential results for pathological voice types. An instructional program based upon synthesized voice signals was developed to facilitate an awareness of prototypical voice types. Eighty phonatory samples representing normal subjects as well as patients with unilateral vocal fold paralysis, vocal nodules, and functional dysphonia were analyzed acoustically on the basis of four measures: average fundamental frequency (F0), jitter, shimmer, and harmonic/noise ratio (H/N ratio). Following training, 29 listeners classified 62% of the phonatory samples on the basis of breathy, hoarse, rough, and normal. Dysphonic severity of rough voices was predicted more successfully by H/N ratio (r2 = .73) than by shimmer (r2 = .43). Dysphonic severity of breathy voices was predicted only by the combined features of less jitter, more shimmer, and lower H/N ratio (r2 = .74). No combination of acoustic variables was successful in the prediction of the hoarse voice type. PMID- 7474971 TI - Activation of lip motor units with variations in speech rate and phonetic structure. AB - The purpose of this study was to describe the activation characteristics of lip muscle single motor units in relation to speech rate and phonetic structure. Repeated experiments were carried out on three adult subjects from whom recordings of lower lip EMG and two-dimensional displacement were obtained. Single motor unit recordings were obtained from the orbicularis oris inferior (OOI), depressor labii inferior (DLI), and mentalis (MENT) muscles. Subjects' tasks involved repeating CV syllables at 1 to 4 syllables per second (syl/sec). The distribution of interspike intervals and corresponding firing rates were obtained on 11 motor units. The firing rates of OOI and MENT motor units increased as syllable rate changed from 1 to 3 syl/sec, but firing rates tended to be equivalent at 3 and 4 syl/sec. DLI and tonic motor units showed little or no modulation in their firing rates with speech rate. Firing rate data and related observations on lip movement and EMG spike count levels suggest that distinct neuromechanical processes control lip movements at low and high speech rates. Both kinematic and EMG data support the expectation that phonetic structure has its greatest effects on lip opening compared to lip closing movements in CV syllables. OOI and MENT activation levels tended to be highest for /p/ productions compared to /w/ and /f/. This may be related to the requirements for complete lip closure and elevated levels of intraoral pressure for production of /p/. PMID- 7474972 TI - Variations across time in acoustic and electroglottographic measures of phonatory function in women with and without vocal nodules. AB - The purpose of this investigation was to identify variations over time in phonatory function of women with and without vocal nodules using acoustic and electroglottographic measures. Subjects were 10 women with vocal nodules (mean age = 22.1, range = 19-25) and 10 women with healthy larynges (mean age = 25.0, range = 18-32). Electroglottographic and audio recordings of speech were obtained for each subject over 3 consecutive days at three target times: morning, afternoon, and evening. Estimates of fundamental frequency (Hz), jitter (msec), shimmer (dB), and signal-to-noise ratio (dB) were made from a 1000 msec midportion of the vowel /a/ produced in a carrier phrase. In addition, a closed to-open ratio was derived from the EGG duty cycle of the same 1000 msec segment and used to estimate timing characteristics of vocal fold vibration. The results showed no significant differences between the groups regarding a pattern of change in the acoustic or the EGG measures across times throughout the day. Furthermore, the experimental group demonstrated significantly lower closed-to open ratios than the control group. With this exception, no other statistically significant differences between the groups were found. PMID- 7474973 TI - Some spectral correlates of pathological breathy and rough voice quality for different types of vowel fragments. AB - This study deals with the relation between listeners' ratings of pathological breathiness and roughness and certain characteristics of the voice spectrum. Two general research questions were addressed: First, which spectral parameters may serve as useful predictors of breathiness and roughness? Second, does the type of speech fragment used for analysis have an effect on the obtained regression model? Listener ratings of breathiness and roughness were obtained for three types of vowel fragments: a vowel onset segment, a mid-vowel (post-onset) segment, and a vowel segment covering the onset and the acoustically more stable post-onset parts. Results indicated that the harmonics-to-noise ratio was the best single predictor of both rated breathiness and roughness, explaining up to 54% of the true rating variance. By combining different predictors, between 75% and 80% of the breathiness variance could be explained for all three types of fragments. For roughness, a strong effect of fragment type was observed, with most variance explained in vowel onset fragments (71%), and least in post-onset fragments (52%). The effect of fragment type was also observed when regression analyses were performed with six predictors based on a factor analysis of the acoustic data. PMID- 7474974 TI - Parameter estimation of labial movements in speech production: implications for speech motor control. AB - Central to theories of speech motor control are estimates on magnitudes of lip activity expressed in terms of central tendency, variability, and interrelatedness. In fact, the tenability of each of two competing theories of motor control for speech production rests solely on the observation of the predicted direction of the correlation coefficient (one positive and one negative) that indexes the relationship of concurrent lip activity. Each theory, however, predicts a relationship that is the complete opposite of the relationship predicted by the other. That is, one theory proposes that the labial system functions on the basis of complementary variation, whereas the other assumes positive covariation, or complementary modulation. In apparent contradiction, each prediction has been observed under laboratory conditions. The explanation for this apparent contradiction resides in the small sample sizes upon which each estimate was based. The minimum number of observations that are necessary to achieve accurate estimates of lip displacement parameters has remained unclear. This paper addresses three fundamental questions: (a) how many observations of on-task behavior are necessary to accurately estimate mean and variance values for the magnitude of upper lip displacement in a speech production experiment?, (b) what is the analogous number of observations for estimating the same values of lower lip displacement (together with the mandible) in the same context?, and (c) how many observations are necessary to accurately estimate the correlation coefficient indexing the relationship of lip displacements during the production of speech? Answers to these questions are accomplished through a review of estimator properties, a Monte Carlo computer simulation, and through laboratory observations.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7474975 TI - Using regions and indices in EPG data reduction. AB - This note describes how dynamic electropalatography (EPG) can be used for the acquisition and analysis of articulatory data. Various data reduction procedures developed to analyze the electropalatographic data are reported. Specifically, these procedures concern two interesting areas in EPG data analysis--first, the novel use of speaker-specific articulatory regions and second, the development of arithmetic indices to quantify time-varying articulatory behavior and reflect reduction and coarticulation. PMID- 7474976 TI - Traditional and phonological treatment for teaching English fricatives and affricates to Koreans. AB - Non-native speakers of English are being seen for accent reduction in speech language pathology clinics although there is little evidence that treatment is effective. Two types of treatment were evaluated. Traditional articulation-based treatment was successful for 2 native Korean speakers in improving English fricatives and affricate production. A second treatment using phonologically based contrasts was also successful for the same sounds with 2 other Korean speakers. Some issues in foreign accent reduction are discussed. PMID- 7474977 TI - A phonologically based analysis of misspellings by third graders with disordered phonology histories. AB - Misspellings evidenced in written responses of third graders during administration of a battery containing 25 true words and 20 nonsense syllable items were analyzed phonologically. Children with histories of disordered phonologies evidenced more phonologically based deviations in their written misspellings than did their peers who had experienced normal phonological development. In addition, the children with disordered-phonology histories relied more on less productive spelling strategies and evidenced poorer phonological awareness skills than their peers with normal-phonology histories. PMID- 7474978 TI - Specific language impairment as a period of extended optional infinitive. AB - English-speaking children with specific language impairment (SLI) are known to have particular difficulty with the acquisition of grammatical morphemes that carry tense and agreement features, such as the past tense -ed and third-person singular present -s. In this study, an Extended Optional Infinitive (EOI) account of SLI is evaluated. In this account, -ed, -s, BE, and DO are regarded as finiteness markers. This model predicts that finiteness markers are omitted for an extended period of time for nonimpaired children, and that this period will be extended for a longer time in children with SLI. At the same time, it predicts that if finiteness markers are present, they will be used correctly. These predictions are tested in this study. Subjects were 18 5-year-old children with SLI with expressive and receptive language deficits and two comparison groups of children developing language normally: 22 CA-equivalent (5N) and 20 younger, MLU equivalent children (3N). It was found that the children with SLI used nonfinite forms of lexical verbs, or omitted BE and DO, more frequently than children in the 5N and 3N groups. At the same time, like the normally developing children, when the children with SLI marked finiteness, they did so appropriately. Most strikingly, the SLI group was highly accurate in marking agreement on BE and DO forms. The findings are discussed in terms of the predictions of the EOI model, in comparison to other models of the grammatical limitations of children with SLI. PMID- 7474979 TI - Speaking rate, articulatory speed, and linguistic processing in children and adolescents with severe traumatic brain injury. AB - Two studies were conducted to examine speaking rate following traumatic brain injury (TBI) in childhood and adolescence. Study 1 focused on longitudinal changes in speaking rate in 9 subjects with severe TBI and their age-matched control subjects. Physical measurements of speaking rate (in syllables/sec) were made from spontaneous speech samples obtained from each subject during three sampling sessions over a 13-month period. Although the average speaking rate of the group with TBI was slower than that of the control group at all three sampling sessions, an examination of the data from individual subject pairs revealed markedly slower speaking rates in only 5 of the 9 subjects with TBI at the final sampling session. The perceptual significance of slowed speaking rates in these 5 subjects was confirmed through subjective ratings by naive listeners. In Study 2, the contributions of two potential causes of slowed speaking rate were explored: reduced articulatory speed and increased pausing believed to be associated with linguistic processing difficulties. It is hypothesized that articulatory speed and linguistic processing speed may contribute independently to slowed speaking rates more than 1 year after TBI. PMID- 7474980 TI - Metrical patterns of words and production accuracy. AB - This investigation examined the influence of metrical patterns of words (syllable stress and serial position) on the production accuracy of 20 children (22 to 28 months). The data were productions of six pairs of individualized two-syllable experimental words that referred to unfamiliar objects. Members of each pair differed only in the placement of stress (e.g., ['soti] vs. [so'ti]). Unstressed syllables were much more likely to be omitted, particularly at the beginning of words. Very few stressed syllables and unstressed second position syllables were omitted. One fourth of the word initial unstressed syllables were omitted. Consonant omissions, though few in number, tended to occur in initial position. Assimilation errors were not influenced by stress or serial position. When segmental errors due to syllable omissions were excluded, other consonant errors were not affected by stress or serial position. These findings indicate that young children's productions of syllables are influenced by the metrical patterns of words. However, the trochaic pattern of English is a statistical tendency, not an absolute constraint on two-syllable words. Metrical pattern also does not affect the consonant accuracy in syllables produced. PMID- 7474981 TI - Discourse comprehension of synthetic speech across three augmentative and alternative communication (AAC) output methods. AB - The purpose of this investigation was to determine the relative effects of three different Augmentative and Alternative Communication (AAC) speech output methods (word, sentence, mixed words and letters) on a listener's ability to summarize paragraph-length texts. Based on previous work on the discourse processing of synthetic speech, a monotonic trend in a listener's ability to accurately summarize texts produced by different output methods was hypothesized (word > sentence > mixed). Thirty able-bodied adults were individually tested over a 2 day period, listening to four texts produced by a DECtalk speech synthesizer at a speech presentation rate of 7.5 wpm. Using a discourse summarization taxonomy developed by Higginbotham, Lundy, and Scally (1993), the experimental hypothesis was confirmed. Word-method listeners produced significantly more accurate renditions of the original texts than sentence-method listeners, who, in turn, did better than mixed-method listeners. Statistically significant differences also were found for the topic familiarity of the texts. The role of the above variables on AAC device comprehension and technology design is discussed. PMID- 7474982 TI - Adult partner-augmented communication input to youth with mental retardation using the System for Augmenting Language (SAL). AB - The primary purpose of this study was to characterize the frequency and nature of augmented input that adult partners provided to 13 youth with mental retardation as they began to use the System for Augmenting Language (SAL). Analyses of youth partner interactions revealed differences in the frequency with which home and school partners provided augmented input and in the manner and style of home and school partners' augmented input, particularly in directiveness and position of lexigram symbols within Utterances. Overall, partners naturally provided augmented input in a manner likely to promote youth's learning of the SAL. PMID- 7474984 TI - Childhood hearing impairment: auditory and linguistic interactions during multidimensional speech processing. AB - Children with mild-severe sensorineural hearing losses often use hearing aids and aural/oral language as their primary mode of communication, yet we know little about how speech is processed by these children. The purpose of this research was to investigate how the multidimensional information underlying accurate speech perception is processed by children with mild-severe hearing impairments. The processing of the auditory and linguistic dimensions of speech was assessed with a speeded selective-attention task (Garner, 1974a). Listeners were required to attend selectively to an auditory dimension (gender of the talker) and ignore a linguistic dimension (word) and vice versa. The hypothesis underlying the task is that performance for the target dimension will be unaffected by what is happening on the nontarget dimension if the dimensions are processed independently. On the other hand, if the dimensions are not processed independently, listeners will not be able to attend selectively and performance for the relevant dimension will be affected by what is happening on the irrelevant dimension (termed "Garner" interference). Both children with normal hearing (N = 90) and children with hearing impairment (N = 40) showed some degree of Garner interference, implying that the dimensions of speech are not processed independently by these children. However, relative to the children with normal hearing, the children with hearing impairment showed normal Garner interference when attending selectively to the word dimension (normally effective at ignoring talker-gender input) and reduced Garner interference when attending selectively to the talker-gender dimension (more effective at ignoring word input). This pattern of results implies that the auditory dimension has a normal strength-of-processing level that makes it normally distracting and that the linguistic dimension has an underdeveloped strength-of-processing level that makes it easier to ignore in children with hearing impairment. PMID- 7474983 TI - Derivation of frequency-gain characteristics for maximizing speech reception in noise. AB - Hearing aid gain-assignment schemes known as "prescriptions" were not designed for fitting hearing aids that modify their frequency responses to reduce background noise interference. Rather, prescriptions were developed for hearing aids having single, fixed frequency responses and aim to optimize speech reception in relatively quiet environments. Even though prescriptions do not apply to noisy conditions specifically, they embody the trade between maximizing speech audibility and maintaining loudness comfort that is critical to frequency gain characteristic selection independent of whether noise is present or absent. The articulation index (AI) was used to examine the extent to which prescriptions' deference to loudness comfort causes them to fall short of maximizing speech spectrum audibility, thereby revealing (roughly) the magnitude of the loudness control built into prescriptions. AIs for speech amplified by an AI-maximizing rule (MAX AI) (Rankovic, Freyman, & Zurek, 1992) and according to several prescriptions were calculated as a function of hearing loss degree and configuration for quiet and noisy conditions. In quiet, AIs for prescriptions were similar to one another when presented with the same audiogram but were drastically smaller than MAX AIs, implying that prescriptions limit speech audibility to a large extent to prevent loudness discomfort. In noise, maximizing the AI required frequency-gain characteristics that were substantially different from prescription-assigned characteristics and that were unique to each noise/audiogram combination. A loudness constraint for the MAX AI scheme was developed to account for the gain discrepancy between prescription AIs and MAX AIs observed in the quiet condition, based on the highest comfortable loudness (HCL) equations presented by Cox (1989) in combination with a loudness model (von Paulus & Zwicker, 1972). The MAX AI scheme with the new loudness control was extended to specify frequency-gain characteristics expected to be optimal for several conditions containing noise, and examples are presented. PMID- 7474985 TI - Temporary and permanent threshold shift caused by hearing aid use. AB - Excessive amplification by hearing aids causes temporary threshold shift (TTS) and permanent threshold shift (PTS). This investigation addressed the question whether it might be possible to predict the eventual amount of PTS caused by excessive amplification from the amount of TTS it causes after a day of hearing aid use. Asymptotic TTS (ATS) to be expected as a result of hearing aid use was predicted for 8 children with sensorineural hearing loss and the predicted ATS was compared with observed permanent deterioration of their thresholds attributed to hearing aid use. There was good agreement between the predicted ATS and observed PTS at 500 to 2000 Hz. It follows that, for prediction of PTS caused by hearing aid use, the mean of the sound levels produced in the ear by the hearing aid is the correct equivalent continuous level (ECL) to use and that the Modified Power Law (MPL) is the appropriate method of adjusting the predictions for sensorineural hearing loss, because these have been shown to be appropriate for prediction of TTS caused by hearing aid use. Predictions of the PTS to be expected for the children that were carried out using the MPL and the mean level as the ECL were in good agreement with the observed PTS at 500 to 2000 Hz, whereas predictions of PTS based on an alternative method of correction for sensorineural hearing loss (the Continuation Hypothesis) were significantly less than the observed amounts. The results of the PTS predictions therefore confirmed the conclusions drawn from the results of the ATS predictions. PMID- 7474986 TI - Physical activity and high density lipoprotein cholesterol. PMID- 7474987 TI - Relationship between strength qualities and sprinting performance. AB - The purpose of this study was to investigate the relationship between strength measures and sprinting performance, and to determine if these relationships varied for different phases of sprint running. Twenty (11 males and 9 females) elite junior track and field athletes served as subjects. Athletes performed maximum sprints to 50 m from a block start and time to 2.5, 5, 10, 20, 30, 40 and 50 m were recorded by electronic timing gates. The resultant forces applied to the blocks were obtained from two force platforms. Twenty-seven measures of strength and speed-strength (absolute and relative to bodyweight) were collected from the height jumped and the force-time curve recorded from the takeoff phase of vertical jumping movements utilizing pure concentric, stretch shortening cycle (SSC) and isometric muscular contractions. Pearson correlation analysis revealed that the single best predictor of starting performance (2.5 m time) was the peak force (relative to bodyweight) generated during a jump from a 120 degree knee angle (concentric contraction) (r = 0.86, p = 0.0001). The single best correlate of maximum sprinting speed was the force applied at 100 ms (relative to bodyweight) from the start of a loaded jumping action (concentric contraction) (r = 0.80, p = 0.0001). SSC measures and maximum absolute strength were more related to maximum sprinting speed than starting ability. It was concluded that strength qualities were related to sprinting performance and these relationships differed for starting and maximum speed sprinting. PMID- 7474988 TI - Energy specificity of rock climbing and aerobic capacity in competitive sport rock climbers. AB - Over the past few years, competitive rock climbing has experienced increased popularity world wide. In 1989, the first six-event World Cup competition was held with all events contested on artificial modular walls. The aim of this study was to determine the extent to which oxidative metabolism is utilized in competitive rock climbing with regard to the climber's maximal O2 consumption (VO2max). VO2max--was measured with two direct triangular protocols: the first from running ("running" VO2max) and the second from pull offs performed with arms and before arms ("pulling" VO2). Moreover, VO2 was also before measured during two competitive climbing routes difficulty quantified 7b on the European numerical scale ranging from 5 to 9. However these routes had different profiles: route 1 was more complex from the informational aspect, holds being smaller and more difficult to see even though the second route was presumed harder from the physical point of view, the holds being bigger but the profile being steeper. The first and the second route involved only 45.6% and 37.7% of the "running" VO2max but 111.6% and 92.3% of the "pulling" VO2max. Heart rates (HR) were equal to 176 bpm and 159 bpm i.e. 85.5% and 77% of maximal HR respectively. Blood lactate collected three minutes after the end of the two ascents were 5.7 mmol.1(-1) and 4.3 mmol.1(-1). The paired "t" test indicated no significant differences in heart rates for the two exercises condition i.e. climbing route. These results suggest that the competitive rock climbing elicit particularly arms since heart rate is high for a relatively low value of VO2.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7474989 TI - Cross-section areas of calf muscles in athletes of different sports. AB - Differences in cross-section areas of calf structures were studied in different sportsmen groups and controls: sprinters, long-distance runners, free-climbers, and sport non active persons. The cross-section areas of calf tissues were established by magnetic resonance imaging. There were no statistically significant differences between the groups in single muscle areas. The only statistically significant difference was found in subcutaneous fat between sportsmen and non active persons. Results show a general adaptive tendency of the muscle to greater activity, without exhibiting sport-group specificity differences. PMID- 7474990 TI - Electromyographic responses at the neuromuscular fatigue threshold. AB - The purposes of this investigation were to: (a) examine the electromyographic (EMG) responses from the vastus lateralis during a continuous ride to exhaustion at the neuromuscular fatigue threshold (EMGFT) and (b) determine what percentage of the power output at VO2max the EMGFT represents. Fifteen adult males (mean +/- SD = 22 +/- 2 years) volunteered to perform an EMGFT test and a continuous ride to exhaustion at the EMGFT on separate days. A subsample of ten subjects completed an incremental cycle ergometer test for the determination of VO2max. The mean (+/- SD) EMGFT for the total sample (n = 15) was 286 +/- 40W and the mean time to exhaustion at the EMGFT was 250 +/- 85 sec. For each subject, the slope of the iEMG versus time relationship during the continuous ride to exhaustion at the EMGFT was significantly (p < 0.05) greater than zero slope. For the subsample (n = 10), the mean EMGFT (296 +/- 45W) was not significantly (p > 0.05) different from the power output at VO2max (282 +/- 47W). These findings did not support the contention that the EMGFT can be sustained without electromyographic evidence of neuromuscular fatigue and indicated that the EMGFT was comparable to the power output at VO2max. PMID- 7474991 TI - Ratings of perceived exertion at maximal exercise in children performing different graded exercise test. AB - The purpose of this study was to examine the rating of perceived exertion (RPE) at maximal exercise in children performing three different graded exercise tests on a treadmill. Eighteen children with a mean +/- SD age of 10.3 +/- 1.4 years volunteered for this study. Each subject performed three different graded exercise test protocols on a treadmill. The protocols consisted of walking only, running only, and combination walk/run. Mean VO2max (ml.kg-1.min-1) during the walking trial (47.3 +/- 5.0) was significantly (p < 0.05) less than during the running (52.1 +/- 4.9) and combination trials (51.9 +/- 5.1). Mean HRmax (bpm) was significantly (p < 0.05) lower during the walking test (195.2 +/- 7.9) compared to the running (202.2 +/- 7.4) and combination tests (202.3 +/- 6.6). Mean RPE at maximal exercise was 18.1 +/- 2.1 in the walking trial, 16.8 +/- 3.0 in the running trial, and 16.8 +/- 2.6 in the combination trial, and was significantly (p < 0.05) greater in the walking trial compared to the other two testing protocols. These data indicate that although VO2max during a walking test is lower, effort sensation is higher compared to protocols that use running as the mode of exercise. The factors involved with RPE selection in this age-group remain to be established. PMID- 7474992 TI - The metabolic responses of slideboard exercise in females. AB - Recently, slideboard exercise training has been introduced as a mode of aerobic training. The purpose of the investigation was to determine the acute cardiovascular and metabolic responses to slideboard exercise. Subjects consisted of 20 healthy females with a mean age of 26.5 +/- 7.9 years, and a VO2max of 40.8 +/- 5.6 ml.kg-1.min-1. The results of the investigation showed that 10 min of a commercially produced slideboard routine elicited mean heart rate (HR) responses ranging from 160 to 168 beats.min-1 and VO2 values of 27.5 to 33.5 ml.kg-1.min-1. A sub-group of 10 subjects was also evaluated while performing sliding exercise at different sliding rates (SR). Subjects performed the standard slide technique at metronome cadences of SR-40, SR-50, and SR-60 slide.min-1. As sliding rate increased so did HR and VO2. The subjects demonstrated mean responses of approximately 55 to 81% of VO2max for the slower to the most rapid sliding rates, respectively. Compared to treadmill exercise at the same oxygen consumption levels, the heart rates for sliding were approximately 15% higher. Slideboard exercise can be an alternative mode of aerobic training. PMID- 7474993 TI - Acute effects of walking on serum lipids and lipoproteins in women. AB - The purpose of this study was to characterize and evaluate the acute effects of walking performed of fairly light (50% VO2max) and moderate (70% VO2max) intensities on serum lipids and lipoproteins in a group of premenopausal (n = 11) and a group of postmenopausal (n = 10) women. Premenopausal women were (x +/- SEM) 34.5 +/- 1.1 years of age, had 22.8 +/- 1.7% body fat and a 2.47 +/- 0.08 l.min-1 VO2max. Postmenopausal women were 54.8 +/- 2.5 years of age, had 37.9 +/- 0.9% body fat and a 2.06 +/- 0.15 l.min-1 VO2max. All subjects walked on a motor driven treadmill at each respective intensity of exercise for a total duration sufficient to expend 350 kcal of energy. Dependent variables included total cholesterol (TC), high-density lipoprotein cholesterol (HDL-C) and its subfractions HDL2-C and HDL3-C, low-density lipoprotein cholesterol (LDL-C), and triglycerides (TG). Blood samples were obtained at baseline (pre-exercise), immediately post-exercise (IPE), and at 24 hours and 48 hours post-exercise. A repeated measures design was employed controlling for diet, menstrual cycle periodicity, natural menopause, and plasma volume shifts. A 2 x 4 ANOVA was used to test for differences among means for each group separately. Significant (p < 0.05) time exercise intensity interactions were found for TC and LDL-C for the premenopausal women. This non-parallel change across exercise intensity condition created significant differences at IPE for both TC and LDL-C. Furthermore, an IPE increase in TG (p < 0.05) was observed.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7474994 TI - A multivariate analysis of kinanthropometric profiles of elite female orienteers. AB - The purpose of this study was to examine the relevance of various kinanthropometric features of elite female orienteers in terms of success at the Student World Orienteering Championships (SWOC)(1992). Participants at this competition were divided into successful (final placing in top 35; n = 12) and less successful (final placing below 35; n = 11) competitors. A non-orienteering reference group was used to enable the identification of any sport specific factors. Measurements of the orienteering group were taken during the week of the SWOC and included 5 skinfold thicknesses, 2 limb girths, 2 bone breadths, 3 proportional lengths and 3 physical tests (grip strength, leg strength, flexibility). Using the results of these, percent adiposity, somatotypes and various anthropometric indices were also obtained. Age and adiposity variables were found to correlate significantly with competitive performance (p < 0.01). Discriminant function analysis successfully distinguished between the successful and less successful orienteers (Wilks' Lambda = 0.603; p < 0.01). There was no difference (p > 0.05) between the orienteering group and the reference group for the majority of the proportionality measures and similarily in the results of the physical tests. Adiposity values were higher in the non-orienteers. In conclusion, body composition measures discriminated between successful and less successful orienteers at a major event at an elite level. The primary difference between the orienteers and the reference group was in body composition rather than linear or proportional measures. PMID- 7474995 TI - The concept of lactate threshold. A short review. AB - The anaerobic threshold (AT) is a widely used tool for investigating aerobic performance characteristics in physiological of pathological conditions. The aim of the present paper is to show that, when the lactate concentration in blood [Lab] is constant in time, regardless of its absolute level, the whole body energy sources for muscular work are entirely aerobic. In fact, [Lab] can remain constant if, and only if, La production is equal to La removal. Since this last is an entirely aerobic process, it can be shown that the net anaerobic energy yield from La production is nil, even if some muscle fibres are indeed producing La at a non trivial rate. These conditions will be defined as "unevenly aerobic" to distinguish them from: (1) the traditional "evenly aerobic" ones, in which the net La production is also zero, but because neither La production nor La removal are significantly increased, and (2) "true anaerobic" conditions wherein La production exceeds La removal and therefore [Lab] increases continuously in time. Comparison of unevenly versus evenly aerobic conditions shows that in the former case the depletion of the glycogen stores is faster in the muscles (or muscle fibres) that are producing La than in those which remove it. Hence the La producing fibres may become crucial in setting the duration (or intensity) of performance. AT, irrespective of its precise mode of assessment, is presumably a measure of the exercise intensity corresponding (or close) to the transition between evenly and unevenly aerobic conditions, thus explaining why AT is a good estimate of the subjects' training status and/or performance capacity. PMID- 7474996 TI - Body composition measurement in elite heavyweight oarswomen: a comparison of five methods. AB - The study was designed to evaluate the range of body composition in elite heavyweight oarswomen as well as the level of agreement between various methods used to measure this variable. Percent body fat was determined at the start of the competitive season by densitometry, taken to represent the reference standard, and measurement of total body potassium, skinfold thicknesses, bioelectrical impedance analysis and body mass index. The athletes were studied within a two week period with all measurements in any individual taken during one morning. We demonstrated a surprisingly large range of percent fat between these oarswomen, 13.6 to 29.3% by densitometry, which was a feature common to all methods. Percent body fat by total body potassium was lower (p < 0.05) while from body mass index higher (p < 0.01) than the reference value from densitometry. Similar methodologies generated significantly different estimates of % fat (SFT1 versus SFT2, p < 0.01 and BIAv versus BIAB, p < 0.01) highlighting the potential problems that may arise with the use of different regression equations to convert primary measurements into % fat. The limits of agreement between various methods were wide and reflect the large variability about the estimated mean bias. Practically this negates the correction of "non reference" values by adding or subtracting the mean difference or bias between the techniques in individuals. These methodological problems need to be considered when setting specific body composition targets for an athlete. PMID- 7474997 TI - Measurement of tissue viability using intravital microscopy and fluorescent nuclear dyes. AB - Intravital microscopy used with fluorescent vital stains provides the opportunity to measure the temporal and spatial extent of tissue injury following disease processes. However, this assumes that prolonged exposure to such dyes does not alter microvascular perfusion or cellular viability. To test this hypothesis, the extensor digitorum longus (EDL) muscle in 24 male Wistar rats, anesthetized with sodium pentobarbital (Somnotal, 65 mg/kg, ip), were prepared for microscopy. The EDL was either bathed continuously (n = 6) in Krebs solution containing bisbenzimide (5 micrograms/ml; labels nuclei of all cells) and ethidium bromide (5 micrograms/ml; labels nuclei of injured cells) or had dyes topically applied 1 hr (n = 4) and 4 hr (n = 4) following dissection of the muscle. Noxious stimuli (i.e., hypoxia:FiO2 of 8-10% (n = 3), 95% ethanol (n = 3), and 2 hr ischemia followed by 90 min reperfusion (n = 4) were used to test the ability of ethidium bromide, when used in conjunction with intravital microscopy, to differentiate injured tissue. Video recordings at the surface of the EDL muscle were made every 30 min for 5 hr from which the number of perfused capillaries was counted (NCper). The numbers of bisbenzimide- and ethidium bromide-labeled nuclei were counted at the surface of the muscle and at two to three additional locations within the muscle (to a maximum depth of approximately 120-160 microns). The average NCper (19.05 +/- 1.7) remained constant over 5 hr, while the number of nuclei stained by bisbenzimide increased linearly with time from an initial value of 1218 +/- 125.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7474998 TI - Recovery of synergistic skeletal muscle function following ischemia. AB - This study tests the hypothesis that if the fast twitch muscles in a synergistic group were more susceptible to ischemia/reperfusion injury, then the slow twitch muscle would compensate functionally during recovery. Rat hindlimb fast twitch gastrocnemius and plantaris muscles and slow twitch soleus muscle were studied. In the experimental (E) group of rats, the right hindlimbs had 2 hr of pressure controlled (300 mmHg) tourniquet ischemia. The masses and the maximal isometric tetanic forces of the three muscles were evaluated at 1, 3, 5, and 7 weeks in E (n = 24) and a control (C) group of rats (n = 24). Gastrocnemius mass and plantaris mass were both reduced (at 1, 3, and 5 weeks and at 1 and 3 weeks, respectively), whereas there were no significant changes in the mass of the soleus. The maximal isometric tetanic forces (N) measured at 1 week of recovery were reduced to 52, 53, and 67% of C values for the gastrocnemius, plantaris, and soleus, respectively. However, at 1 week the normalized isometric tetanic forces (N/g) for all three muscles were reduced to 66-69% of the C values. By Week 3, the tetanic forces (N or N/g) of all muscles had recovered to control values. On average, the gastrocnemius, plantaris, and soleus muscles of the C groups contributed 68, 24, and 8%, respectively, of the total synergistic tetanic force. These values were unaffected by ischemia. From this experiment, it is concluded that 2 hr of tourniquet ischemia resulted in a selective decrement in mass of the fast twitch muscles.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7474999 TI - Development of circulatory and metabolic shock following transient portal triad occlusion. AB - Liver ischemia is purposefully induced by portal triad occlusion (PTO) in several clinical situations including liver surgery for trauma, tumor, and transplantation. Despite significant morbidity from PTO, the hemodynamic and metabolic effects of PTO have not been evaluated relative to duration of ischemia. We investigated this using a total hepatic ischemia model. Rats received isoflurane anesthesia, carotid artery and jugular vein cannulation, and serial measurements of cardiac output (CO), mean arterial pressure (MAP), heart rate (HR), central venous pressure (CVP), stroke volume (SV), systemic vascular resistance (SVR), superior mesenteric artery blood flow (SMAF), intestinal vascular resistance (IVR), pH, pCO2, pO2, lactate, glucose, hematocrit (HCT), white blood cell count (WBC), and total neutrophils. Each group received 0, 15, 30, 45, or 60 min of PTO followed by 2 hr of reperfusion. All sham ischemia animals remained hemodynamically stable throughout the study. However, in the ischemic groups, there were significant time-dependent decreases in MAP, HR, CO, CVP, SV, SMAF, and pH, and increases in SVR, IVR, HCT, and lactate, while pCO2, pO2, glucose, and WBC remained stable. All of the ischemic animals survived except those that received 60 min of PTO. In this group, all of the animals survived the ischemic period; however, only one animal survived beyond 60 min of reperfusion. These data demonstrate a time-dependent circulatory and metabolic shock following PTO heralded by intestinal venous pooling and loss of intravascular fluid, and culminating in death. Careful hemodynamic monitoring and restoration of blood volume in the trauma patient may reduce morbidity and mortality. PMID- 7475000 TI - Reconstituted high-density lipoprotein reduces LPS-stimulated TNF alpha. AB - Reconstituted high-density lipoprotein (rHDL), an artificial lipoprotein consisting of apolipoprotein A-I and phosphatidylcholine (1:150, molar ratios) dose-dependently reduces lipopolysaccharide (LPS)-induced tumor necrosis factor alpha (TNF) production in in vitro, ex-vivo, and in-vivo model systems. In an in vitro whole blood assay, rHDL (1 mg/ml) added concomitantly with LPS increased cellular resistence to LPS stimulation approximately 1000-fold. Even with extremely high levels of LPS (10 micrograms/ml), rHDL > or = 0.5 mg/ml caused > 50% decrease in TNF production. Preincubation of rHDL with LPS was not required for activity. rHDL (> or = 1 mg/ml) reduced TNF production by 50% even when added to cultures 2 hr after their stimulation with LPS (10 micrograms/ml). In an ex vivo study, rabbits were infused with rHDL at doses of 25, 50, and 75 mg/kg. Blood was drawn and stimulated with LPS ex vivo and bioactive TNF was assessed using the L929 cytotoxicity assay. Fifteen minutes after rHDL infusion, there was a significant difference in ex-vivo-induced TNF activity between groups (750 +/- 160, 170 +/- 40, 80 +/- 30, 60 +/- 30 pg TNF/ml, for the control, 25, 50, and 75 mg/kg rHDL dose groups, respectively; P < 0.0001). The duration of ex-vivo TNF inhibition was dependent on the dose of rHDL. Even at 2 hr, rHDL showed a pronounced TNF inhibition (control: 950 +/- 120 pg TNF/ml; 75 mg/kg: 140 +/- 60 pg TNF/ml). Further studies showed that a prophylactic infusion of rHDL diminished LPS-induced TNF production in a rabbit endotoxemia model.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7475001 TI - Role of leukocyte activation in patients with venous stasis ulcers. AB - Alteration in leukocyte activation has been implicated as an etiological factor in the development of chronic venous stasis ulcers (CVSU). The purpose of this study was to determine differences in expression of cell surface activation markers on circulating leukocytes and systemic, soluble, serum cytokine levels between healthy controls and patients with CVSU. Twenty-three patients were separated into two groups. Group I consisted of 12 healthy, adult, age-matched male patients with no venous disease. Group II consisted of 11 adult male patients with CVSU who underwent air plethysmography (APG) and duplex scanning to determine the severity of venous insufficiency. All patients had measurements of systemic, serum-based, soluble IL-1 beta, IL-2, IL-6, TNF-alpha, and beta 2 microglobulin levels. Using fluorescence flow cytometry, we measured the percentage of lymphocytes (CD3), monocytes (CD14), and granulocytes (CD15) expressing various cell surface activation markers. By APG and duplex scan, all group II patients exhibited venous insufficiency, with a mean venous filling index of 6.9 +/- 3.9 sec. Relative to group I, group II patients demonstrated a decreased expression of the CD3+/DR+ (13.3 +/- 1.5, P < or = 0.01) and CD3+/CD38+ (31.1 +/- 2.1, P < or = 0.04) markers on T-lymphocytes and an increased expression of CD14+/CD38+ (99.6 +/- 0.2, P < or = 0.008) markers on monocytes. Circulating neutrophils showed no evidence of activation. In addition, a significant elevation in the T-helper to T-suppressor ratio (2.9 +/- 0.6, P < or = 0.0001) between groups I and II was observed.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7475002 TI - Experimental tracheal replacement using a revascularized ileal segment for transplantation in rats. AB - The clinical field of tracheal reconstruction is still the subject of intensive investigation. Numerous attempts to replace extensive circumferential tracheal defects have failed to demonstrate long-lasting reliable results. Commonly used alloplastic prostheses are missing a mucosal barrier which clears mucus secretions and protects against contamination by infectious organisms. Tissue granulation and formation of scar strictures at the suture lines are common sequels and mainly contribute to graft failure. We hypothesized that the small bowel could adapt and comply with the functional and physiological requirements of the respiratory tract. To further clarify this study objective, a "two-step" surgical procedure was carried out on LEW inbred rats. First, an ileal segment 6 cm in length was dissected from the intestinal continuity. The divided segment was then sidepassed in a Roux-en-y fashion. The exposed segment was stabilized by a ring-enforced PTFE (polytetrafluoroethylene) prosthesis which was cut open and wrapped around the antimesenteric outside surface of the bowel segment. After 21 days the alloplastic prosthesis was completely incorporated into the adjacent small bowel tissue. Afterward, a tracheal resection of 10 cartilage rings was performed in a second syngeneic animal and the composite graft prepared in animal one was interposed for reconstruction in animal two. In a series of 10 successful experiments, air tightness and a normal respiratory tract without signs of mucus congestion could be confirmed. A gap-free epithelial lining at the suture lines reliably prevented bacterial contamination, subsequent tissue granulation, and formation of scar stricture within the graft.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7475003 TI - Skeletal muscle ventricles: improved left ventricular performance associated with a prolongation of ventricular relaxation. AB - Skeletal muscle ventricles (SMV) have been shown to pump blood effectively in both acute and chronic models. In this study SMVs were placed in circulation in series with the descending thoracic aorta to function as diastolic counterpulsators. The SMVs were stimulated to contract at a 1:2 ratio with the heart. The R-wave delay and the burst duration were varied, both as a percentage of the total R-R' interval to determine the effect of the SMV on left ventricular (LV) relaxation in relation to other parameters of LV function. Optimal hemodynamic benefit was obtained using a 40% R-wave delay and a 30% burst duration. At these settings, the time constant of LV relaxation (tau) increased from a control value of 29.1 +/- 1.6 to 39.8 +/- 2.1 msec (P < 0.001). The tension time index decreased from a control value of 15.24 +/- .31 to 13.88 +/- .7 mm Hg-sec-1 (P < 0.001). The endocardial viability ratio, an indicator of myocardial blood flow, improved from 1.12 +/- .11 to 1.55 +/- .16 (P < 0.001) and LV dP/dt increased from 1555 +/- 180 to 2320 +/- 210 mm Hg-sec-1 (P < 0.05). In conclusion, this study demonstrates SMV diastolic counterpulsation results in improved LV performance which is associated with a prolongation of LV relaxation. A 40% R-wave delay interval and a 30% burst duration were the stimulation parameters which benefitted myocardial function optimally in this study. PMID- 7475004 TI - Prolonged survival of vascularized limb tissue allografts by donor irradiation. AB - The clinical feasibility of transplantation of a vascularized limb tissue allograft depends upon reducing immunosuppression and its associated toxicity for the graft recipient. Donor or allograft irradiation would eradicate allogeneic marrow and provide a possible alternative or beneficial addition to host immunosuppression. The effect of irradiation on survival of limb tissue allograft was investigated in this study. In a rat model, knee allografts consisting of bone, cartilage, and soft tissues were transplanted across a strong histocompatibility barrier by femoral vascular microanastomoses. The grafts were harvested 1 and 2 weeks postoperatively for histologic and immunologic assays. Gamma irradiation of the graft immediately before transplant decreased graft rejection slightly. However, total body irradiation of the donor followed by a "waiting period" of 2 or more days before transplant significantly delayed rejection. After such a 6-day and a 2-day pretreatment protocol, the host cellular immune responses were not observed 1 and 2 weeks after transplantation and eventual progression toward graft rejection coincided with emergence of host antibody production. Rejection of vascularized limb tissue allografts after donor irradiation thus appeared to be mediated primarily through the humoral pathway. We conclude that donor irradiation is a potentially useful adjunct in prolonging survival of vascularized limb tissue allografts. PMID- 7475005 TI - Significance of hyaluronic acid for evaluation of hepatic endothelial cell damage after cold preservation/reperfusion. AB - Serum hyaluronic acid (HA) levels during or after liver transplantation can reflect graft viability in animal models. Decreased HA clearance in nonviable grafts seems to reflect hepatic endothelial cell damage induced by cold preservation/reperfusion. Therefore, we examined the relationship between HA clearance and endothelial cell damage, using an isolated perfused rat liver (IPRL) model. The grafts were separated into four groups, according to cold preservation time: minimal storage ( CONTROL: n = 9), 4-hr preservation (n = 9), 5-hr preservation (n = 8), 6-hr preservation (n = 9). After cold storage, grafts were put on the recirculating IPRL system and then reperfused for 120 min with 37 degrees C oxygenated Krebs-Henseleit buffer, containing HA and sodium taurocholate. To examine the function of sinusoidal endothelial cells and hepatocytes, serial measurements of HA, total bile acids, and conventional parameters in the perfusate were taken. After the perfusion, a trypan blue exclusion test was done to assess the microscopic sinusoidal lining cell (SLC) damage. HA clearance from the perfusate showed a preservation time-dependent decrease which clearly distinguished between 4- and 6-hr preserved grafts. Trypan blue uptake ratio of SLC increased in accordance with preservation time. HA clearance at 120 min after reperfusion showed a highly significant correlation with histologically assessed SLC damage. These results suggest that HA is useful to evaluate the extent of endothelial cell damage after cold storage and reperfusion. The significance of HA as a predictor of graft viability is also confirmed. PMID- 7475006 TI - Fibronectin on the surface of biliary drain materials--a role in bacterial adherence. AB - The present study deals with the demonstration of deposited fibronectin (Fn) on the surfaces of implanted biliary drain materials and the role of deposited Fn in promotion of bacterial adherence. Rubber pieces that had been implanted in the biliary tracts of rats for 4 weeks were retrieved and the following approaches employed for further investigations: (1) adherence of [methyl-3H]thymidine labeled Escherichia coli to implanted and unimplanted rubber pieces; (2) blocking the adherence of radiolabeled bacteria with anti-Fn antibodies; (3) detection of deposited Fn by 125I-labeled anti-Fn IgG; and (4) immunoblotting of the surface eluate from implanted rubber pieces. The results show that in the presence of serum, plasma, or bile, the number of E. coli cells adherent to implanted rubber pieces was 10 times higher than that adherent to the unimplanted pieces (P < 0.001) and that the adherence was reduced by pretreatment of implanted pieces with anti-Fn antibodies. Furthermore, the implanted pieces appeared to have a high affinity for 125I-labeled rabbit anti-Fn IgG rather than the 125I-IgG without anti-Fn fraction. Fn was also found in the surface eluate of implanted pieces by immunoblotting of the eluate. The results in the present study suggest that Fn may be involved in implant-associated infections in the biliary tract. PMID- 7475007 TI - Fibrin glue and conventional sutured vasal anastomosis in the rat. AB - The present study was undertaken to evaluate the use of fibrin glue (Tisseel, Immune U.S., Inc) for vasovasostomy and to compare this technique to conventional sutured vasovasostomy. Thirty immature Sprague-Dawley rats, weighing from 60 to 80 g, were used in this study. A conventional one-layer sutured anastomosis (Ethilon 10-O) in 10 rats was compared to a fibrin glue technique of vasal anastomosis (10 rats). The fibrin glue technique was performed without sutures and was unstented. The biological glue was utilized to seal both ends of the vas. The contralateral vas was ligated with 5-O prolein. In the control group (10 rats) the left vas was ligated in the same way, and only the contralateral vas was explored. After 2 months, one male and two female rats were placed in a cage for a further 2 months. At the end of this period, the fertility rate was 80% (n = 8) of the control group, 60% (n = 6) of the conventional anastomosis, and 70% (n = 7) of fibrin glue groups, respectively. The testes and vasal specimens were evaluated for the presence of sperm granuloma, and histologic studies were performed. The incidence of sperm granuloma after vasovasostomy was 20% (n = 2) for the fibrin glue group and 30% (n = 3) for the sutured group. The sperms were seen in the proximal and distal side of the vasal anastomosis in 10 rats in the control group, in 8 in the glue group, and in 8 in the conventional sutured anastomosis group.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7475008 TI - Acute effects of insulin-like growth factor I on interorgan glucose and lactate flux in protein-catabolic dogs. AB - Insulin-like growth factor-I (IGF-I) is a potent protein-anabolic hormone with a glucose-lowering effect and is therefore a possible agent for treating catabolic patients. In this study we investigated the effect of recombinant human (rh) IGF I on the interorgan flux of glucose under hypo- and normoglycemic conditions in catabolic, anaesthetized, and catheterized dogs. We administered a primed (40 micrograms/kg) continuous (1.5 micrograms.kg-1.min-1) infusion of rhIGF-I (Kabi Biopharma, Stockholm, Sweden) for 180 min together with either a saline (0.9% NaCl) or an amino acid solution (2.2 mg AA.kg-1.min-1 solution of Vamin, Kabi Nutrition, Stockholm, Sweden). RhIGF-I administration lowered plasma glucose levels for approximately 50% of the baseline (P < 0.001) and stimulated glucose uptake from skeletal muscle about twofold (P < 0.01), but did not modify glucose balances across the gut and liver. The same effects were found when infusing rhIGF-I together with AA. A co-infusion of rhIGF-I and glucose to maintain normoglycemic conditions stimulated glucose uptake from skeletal muscle by about fivefold (P < 0.001) and glucose uptake across the gut by about 50%, but reduced the hepatic glucose liberation (-65%; P < 0.01). The rhIGF-I infusion did not alter arterial lactate levels, but stimulated lactate release from skeletal muscle (P < 0.05) and lactate uptake across the liver (P < 0.05). We conclude that rhIGF-I reduces plasma glucose levels mainly by stimulating glucose uptake across skeletal muscle.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7475009 TI - Sterilization of partially demineralized bone matrix: the effects of different sterilization techniques on osteogenetic properties. AB - Transplantation of allogenic bone requires the thorough examination of donors as well as the careful processing and storage of samples in order to minimize potential infection. Other problems associated with allogenic transplants such as low osteoinductive properties and immunological reactions led to the development of partially demineralized bone matrix (PDBM). This highly osteogenic bone extract is largely free of antigens and easy to produce. However, in order to exclude the potential risk of infection, PDBM should be sterilized prior to implantation. It was the purpose of this study to investigate the influence of various sterilization techniques on the osteoinductive properties of PDBM. Seventy-six drill defects with a diameter of 0.6 cm in the tibia of 11 Merino sheep were filled with PDBM as well as autogenic or allogenic cancellous bone. Prior to implantation the PDBM was sterilized using autoclavation, gamma irradiation, ethylene oxide, or ethanol. Twelve empty drill holes served as controls. The extent of new bone formation was ascertained by histological, fluorescent-optical, and microradiographical examinations 3 and 6 weeks postoperatively. Furthermore, the amount of newly formed bone was measured quantitatively. Apart from autoclaved PDBM, all matrix grafts showed excellent new bone formation after sterilization, exceeding the results of allogenic cancellous bone. PMID- 7475010 TI - Integrating an acute geriatric teaching service with a general medicine program in a university hospital. PMID- 7475011 TI - Telephone access to a university HIV/AIDS clinic. PMID- 7475012 TI - Prehospital blunt traumatic arrest: survival and analysis. PMID- 7475013 TI - Aspirin for prevention of myocardial infarction. PMID- 7475014 TI - Loss prevention case of the month. He said--they said. PMID- 7475015 TI - Life-threatening hemolysis following a brown recluse spider bite. PMID- 7475017 TI - Breast cancer: a significant public health concern. PMID- 7475016 TI - Calciphylaxis. PMID- 7475018 TI - Managed care: lessons from the California experience. PMID- 7475019 TI - The expanded role of coronary angioplasty during pregnancy. PMID- 7475020 TI - Bronchiolitis obliterans organizing pneumonia: a community hospital experience. PMID- 7475021 TI - "Back mice"--a prevalence study. PMID- 7475022 TI - Accelerated progressive massive fibrosis and mica dust. PMID- 7475023 TI - A man with fever and arthralgias. PMID- 7475024 TI - Sustained monomorphic ventricular tachycardia: a case of cardiac sarcoidosis. PMID- 7475025 TI - Delay is deadly. PMID- 7475026 TI - Managed care revisited. PMID- 7475027 TI - Validation of daily self-reported alcohol consumption using interactive voice response (IVR) technology. AB - OBJECTIVE: This study assesses the validity of daily self-reported drinking data obtained using an automated touch-tone interactive voice response (IVR) system. METHOD: Subjects (N = 30) reported alcohol consumption daily for 28 days using the IVR system. Concurrently, breath and saliva samples were obtained each night for objective determination of blood alcohol concentrations (BACs). Partners living with the subjects provided collateral reports daily. Retrospective drinking records were obtained from both partners at the outset of the study and from the target subjects at the end of the study, using timeline follow-back procedures referencing the target subjects' drinking over the previous 28-day period. RESULTS: Subjects reported drinking on 55.2% of the 840 possible subject days, and positive BAC readings were obtained on 25.9% of these days. The overall correlation between self-report and measured BAC was .72. Within-subject correlations between daily IVR reports and measured BACs ranged from -.07 to .92, with a mean of .57. The correlations between self-reported drinking and the collateral reports ranged from .18 to 1.0, with a mean of .89. Correlations between the daily self-reports and the timeline follow-back records obtained at the end of the study ranged from -.22 to .96, with a mean of .51. CONCLUSIONS: IVR technology provides an innovative, user-friendly methodology for obtaining valid measures of daily alcohol consumption. The validity of these measures may be differentially highest for frequent, heavy drinkers, a group for whom traditional assessment methods often produce the most biased underestimates. PMID- 7475028 TI - Cycles of alcohol dependence: frequency-domain analyses of daily drinking logs for matched alcohol-dependent and nondependent subjects. AB - OBJECTIVE: This study was undertaken to evaluate possible differences in the patterns of daily alcohol use, in terms of periodicity or rhythmicity of consumption, between alcohol-dependent and nondependent drinkers otherwise matched on the basis of quantity and frequency of alcohol use. METHOD: Frequency domain time-series analysis was applied to daily drinking logs of six alcohol dependent and six nondependent male subjects matched for consumption quantity and frequency who had participated in a larger 112-day study of alcohol consumption patterns using an innovative touch-tone telephone method of daily self-report. Spectral analysis partitioned variance in the drinking logs among statistically independent frequency cycles of relatively heavier and lighter drinking. The percentage of variance accounted for by fast (< 4 days), medium (7 +/- 3 days) and slow (> 10 days) cycle periods was compared for diagnostic group differences. RESULTS: Effective matching precluded consumption quantity, frequency or variability differences between the groups. Spectral analysis of the time-series data revealed group differences in the rhythmicity, or cyclical characteristics, of the drinking logs. The drinking logs of alcohol-dependent subjects exhibited more pronounced weekly cycles, whereas drinking logs of nondependent subjects fluctuated over longer periods. CONCLUSIONS: Temporal patterns of regular alcohol use may be an important behavioral variable influencing the seriousness of consequences associated with drinking, which may partly elucidate the relatively weak relationship between dependence symptomatology and consumption quantity frequency. PMID- 7475029 TI - Reliability of individual diagnostic criterion items for psychoactive substance dependence and the impact on diagnosis. AB - OBJECTIVE: Reliability of diagnostic criterion items for psychoactive substance dependence and the impact of each on the reliability of the diagnosis were analyzed. METHOD: As part of a reliability study for a new interview developed for the multisite Collaborative Study on the Genetics of Alcoholism (COGA), data were collected from both within-center and across centers. The impact of each diagnostic item on the reliability of the substance dependence diagnosis was studied by forcing each item to be reliable one at a time and recomputing the kappa statistic for the diagnosis. RESULTS: Findings indicated that the majority of individual diagnostic criterion items were reliable; 87% and 81% were in the fair or better range of reliability for the within- and cross-center studies, respectively. Individual kappa estimates were statistically similar for the two studies. Reliability findings for two classes of substance, alcohol and cocaine, were good, while those for stimulants were less satisfactory. CONCLUSIONS: Forcing items one at a time to be reliable did not affect reliability of the overall substance dependence diagnosis, because more than one criterion item changed from Time 1 to Time 2. Because no single item was influential, weighting criteria equally, as is done in the DSM and ICD classification systems, appears to be a reasonable approach. PMID- 7475030 TI - Role of alcohol in traffic crashes involving women: passenger car fatalities in northern Sweden. AB - OBJECTIVE: Since increased alcohol consumption and increased driving among women may lead to a growing mortality due to drunken driving among women, the study analyzed gender differences among traffic fatalities. METHOD: Traffic fatalities (597 victims, 159 women) in northern Sweden were investigated, using autopsy and police reports, for a 10-year period, 1980-1989. RESULTS: The incidence of inebriated female driver fatalities was 0.51 per 100 million km (men, 0.66). Eighty-six percent of the female inebriated drivers (98% of men) and 68% of female sober drivers (78% of men) initiated the crash. Blood alcohol was detected in 10% of the women (men, 32%) with a mean blood alcohol concentration of 1.1 g/kg (men, 1.9 g/kg). Of the women, 13% had liver steatosis (men, 28%). The proportion of victims with liver steatosis increased with increasing blood alcohol concentration. Only one case of liver cirrhosis was found (a man). The majority of the inebriated victims were killed from May through October (women, 73%; men, 76%), and from Fridays through Sundays (women, 87%; men, 70%). Only 27% of the inebriated women crashed between 9 PM and 6 AM, compared to 62% of the men. The body location of fatal trauma was similar in men and women and was not influenced by alcohol. CONCLUSIONS: Female traffic fatalities differ from those of men in several respects. No evidence was found for an increase in the number of women in alcohol-related traffic fatalities in Sweden over the 10-year period studied. Drunken driving and alcohol abuse in traffic is still mainly a male problem. PMID- 7475031 TI - Demographic characteristics, drinking patterns and drink-driving behavior of injured male drivers and motorcycle riders. AB - OBJECTIVE: The demographic characteristics, usual drinking and drink-driving patterns, incidence of problem drinking, precrash drinking behavior and attitudes towards drink-driving of crash-involved male drivers and riders are described and related to blood alcohol concentration (BAC) on admission to hospital. A better understanding of these issues will enable drink-driving countermeasures to target more specifically those individuals who are most at risk of being killed or injured in alcohol-related road crashes. METHOD: Between June 1985 and April 1987 interviews were conducted with a sample of 302 male drivers and motorcycle riders admitted to the Royal Adelaide Hospital in Adelaide, South Australia. RESULTS: With one exception, the likelihood of having a high BAC (i.e., 80 mg/dl or above) did not differ with demographic profile. As BAC increased, there was a significant increase in: various indices of quantity and frequency of drinking; beer being the preferred beverage; percentages drinking alone, in a hotel, in a vehicle and for various less socially acceptable reasons; frequency of drink driving; likelihood of previous drink-driving suspension; and, more liberal attitudes towards drink-driving. About 25% of those with a BAC of at least 150 mg/dl were probably experiencing alcohol-related problems prior to the crash, compared with only a very small proportion of those with lower BACs. Precrash drinking most commonly involved drinking in a hotel, drinking with friends and drinking beer, with no significant differences between BAC groups. CONCLUSIONS: The results suggest that usual drinking and drink-driving patterns, as well as attitudes to drink-driving, become more extreme as the BAC of male crash-involved drivers and riders increases. PMID- 7475032 TI - Incidence and characteristics of spinal cord injured patients with a family history of alcoholism. AB - OBJECTIVE: Of the many factors that affect the occurrence of and rehabilitation from spinal cord injury, alcohol use and abuse are particularly prominent. Persons with a family history of alcoholism are at greater risk for alcohol abuse and alcoholism. Preliminary data show a much higher than normal proportion of these persons in the spinal cord injured population. If these data are reliable, they may show that family history of alcoholism is a premorbid or comorbid factor to the occurrence of accidents that results in catastrophic injuries. Our investigation represented an evaluation of this premise. METHOD: Male volunteer subjects (N = 100) were selected from inpatient and outpatient populations of a Veterans Affairs spinal cord injury service. Subjects received a comprehensive interview and completed two standardized questionnaires and a general information form designed to evaluate their past and present use of alcohol and the use of alcohol by their parents and relatives. RESULTS: The incidence (45%) of spinal cord injured patients with a family history of alcoholism was over four times that found in the general population. Spinal cord injured subjects with a family history of alcoholism reported significantly more problems with alcohol before, at the time of, and after their injury. CONCLUSIONS: The data suggest a relationship between the predisposition to alcoholism and accidents that result in catastrophic injury. PMID- 7475033 TI - The role of stress and family relationships in mediating problem drinking and fathers' personal adjustment. AB - OBJECTIVE: To test a stress process model in which family stress (negative life events) and fathers' family system resources (marital adjustment and positive father-child relationships) were evaluated as mediators of the relationship between problem drinking (fathers' and mothers') and fathers' personal adjustment. METHOD: Structural equation modeling was used to analyze data from fathers and mothers in 93 two-parent families with early adolescent children. RESULTS: Fathers' problem drinking contributed marginally to family stress and directly to fathers' diminished personal adjustment. Family stress was related to reduced marital adjustment and personal adjustment. Mothers' problem drinking contributed only to less positive father-child relationships. CONCLUSIONS: Stress process models for fathers and mothers differ; in particular, family relationships do not appear to play a significant mediational role for fathers whereas they do for mothers. Interventions for symptomatic fathers might best concentrate on alleviating problem drinking and extrafamilial sources of stress. PMID- 7475034 TI - Stress, vulnerability and adult alcohol relapse. AB - OBJECTIVE: Alcoholics experiencing highly threatening or chronic psychosocial stress following treatment are more likely to relapse than abstaining individuals not experiencing such stress. Expanding upon this stress-relapse hypothesis, we predicted that individual risk and protective characteristics would contribute to vulnerability to relapse in alcoholic men confronted with significant life adversity. The present investigation examined the relationship between psychosocial vulnerability and return to drinking. METHOD: A group of abstinent male alcoholics (N = 67) who experienced marked life adversity that posed a severe and/or chronic threat participated in this study. Men completed a psychosocial assessment first as an inpatient in treatment for alcohol dependence, and again at 3 months and 1 year following discharge. RESULTS: Among alcoholic men exposed to severe psychosocial stressors, those with higher composite psychosocial vulnerability scores were more likely to subsequently relapse than those with lower vulnerability scores. Additionally, men who improved in psychosocial functioning following treatment had better outcomes than men whose vulnerability increased. In particular, coping, self-efficacy and social support most consistently predicted relapse among this sample of severely stressed abstaining alcoholics. CONCLUSIONS: These findings supported the stress vulnerability model of relapse. Results indicated that improvement in psychosocial domains (e.g., coping skills, social networks, perceived ability to tolerate relapse-risk situations) enhanced the ability of these men to remain abstinent despite severe stress. This study highlights the importance of cognitive and behavioral interventions for increasing improvement in these psychosocial domains. PMID- 7475035 TI - Stress and alcohol: the moderating effect of chronic stress on the acute stress- intoxication relationship. AB - OBJECTIVE: The present study evaluated whether chronic stress levels moderated the impact of laboratory stressors on subjective and behavioral responses to alcohol. METHOD: Healthy volunteers (N = 60; 30 male) completed measures of background stress levels (e.g., major life events). In addition, subjects were exposed to two laboratory stressors (i.e., cold pressor or film stressor task) or a control condition after consuming a 0.7 g/kg dose of alcohol. RESULTS: Regression analyses showed that the combination of high background stress levels and exposure to a lab stressor reduced two measures of perceived intoxication (i.e., Sensation Scale, Visual Analog Intoxication Scale). CONCLUSIONS: These data are consistent with a biobehavioral model of alcohol use where acute and chronic stressors are associated with a diminished response to alcohol. The possible mechanisms that may underlie this sobering effect include stress-related cognitive deficits and situation specific tolerance associated with high chronic stress levels. PMID- 7475036 TI - Gender roles and alcohol abuse across the transition to parenthood. AB - OBJECTIVE: The transition to parenthood has been occurring within the context of the increasing labor force participation of women and potential stress-related costs of "nontraditional" as well as "traditional" gender role choices of new parents. This study addressed the extent to which gender role-related stressors lead to increased problem drinking across the transition to parenthood. METHOD: Fifty-five married couples expecting their first child were obtained from seven Chicago area hospitals and surveyed with a mailed questionnaire at two points in time: (1) during the second trimester of pregnancy and (2) at 6 months following childbirth. The study examined both wives' and husbands' changes in problem related drinking by comparing problem drinking during the 6 months prior to pregnancy (retrospectively assessed at Time 1) with drinking during the 6 months following childbirth using a modified form of the Michigan Alcoholism Screening Test. In addition, the study contrasted problem drinkers with nonproblem drinkers following parenthood with regard to role overload (overwhelming demands from work and family roles), role deprivation (loss of work role following parenthood) and role deterioration (decreased support from role relationships). RESULTS: The data show increased problem drinking for both mothers and fathers but fail to support either role overload or role deprivation hypotheses. However, the data are congruent with the role deterioration perspective. CONCLUSIONS: If these findings are replicated, future studies of gender roles and alcohol abuse should focus greater attention on the perceived quality of social roles beyond noting their presence or absence. PMID- 7475037 TI - Changes in drinking patterns among whites, blacks and Hispanics, 1984-1992. AB - OBJECTIVE: This article reports a longitudinal study of drinking patterns among whites, blacks and Hispanics between 1984 and 1992. METHOD: A probability sample including 1,777 whites, 1,947 blacks and 1,453 Hispanics in the U.S. adult household population was interviewed in 1984. In 1992 a subsample consisting of 788 whites, 723 blacks and 703 Hispanics was reinterviewed. Interviews averaging 1 hour in length were conducted in respondents' home by trained interviewers. RESULTS: Abstention increased in all groups with the exception of Hispanic women, among whom it remained stable. Heavy drinking decreased only among white men (from 19% to 12%). Among men, the incidence of heavy drinking was 7% among whites, 10% among blacks and 17% among Hispanics. The stability of heavy drinking was greater among black (51%) and Hispanic men (43%) than among white men (32%). The best predictor of drinking in 1992 was drinking in 1984. CONCLUSIONS: Reductions in heavy drinking observed among whites were not observed among blacks and Hispanics. The greater stability of heavy drinking in the minority groups helps to explain higher rates of problems reported in the literature for these two groups. Prevention efforts targeting drinking and heavy drinking among blacks and Hispanics must be renewed and intensified. PMID- 7475038 TI - Changes in alcohol consumption resulting from the elimination of retail wine monopolies: results from five U.S. states. AB - OBJECTIVE: The role of publicly owned and operated retail alcohol monopolies is currently under debate in many Western and former Eastern-bloc countries. We studied the effects of privatizing wine sales in five U.S. states. METHOD: Data on monthly sales of alcoholic beverages were collected for each of the five states, for all states bordering each of the five states and for the U.S. as a whole for the period from 1968 through 1991. Beer and spirits sales data were collected for comparison with wine sales. A quasiexperimental interrupted time series design was used, including comparison groups consisting of border states, all other U.S. states and beer and spirits sales within the focal state. Box Jenkins time-series statistical modeling was used to control for intra- and cross series dependencies and to estimate the net effect of privatization on wine sales. RESULTS: After controlling for both nationwide and state-specific trends, we found significant increases in wine sales after privatization of 42% in Alabama, 150% in Idaho, 137% in Maine, 75% in Montana and 15% in New Hampshire. The increases in liters of pure ethanol per year in the form of wine were 621,000 in Alabama, 432,000 in Idaho, 364,000 in Maine, 363,000 in Montana and 171,000 in New Hampshire. CONCLUSIONS: The structure of the retail alcohol distribution system has a significant effect on alcohol sales. We recommend that the social costs associated with increased alcohol use be carefully considered before such major policy changes are contemplated. PMID- 7475039 TI - An investigation of gender differences in alcohol-related aggression. AB - OBJECTIVE: The majority of the research on alcohol-related aggression has been conducted on men. This bias has persisted despite mounting evidence indicating gender differences in aggression. As such, the purpose of this study was to investigate gender differences in alcohol-related aggression. METHOD: Subjects were 64 men and 64 women who competed against either male or female fictitious opponents on a modified version of the Taylor aggression paradigm in which electric shocks were received from and administered to a fictitious opponent during a competitive task. Aggression was operationalized as the intensity and duration of the shocks selected by the subjects. Subjects were assigned to either an alcohol, a placebo or a sober group. RESULTS: Alcohol increased both shock intensity and duration in men; however, it only increased shock duration in women. Men behaved more aggressively toward men, whereas women displayed equal levels of aggression toward both genders. Alcohol expectancies did not appear to have an effect on aggressive behavior for either gender. CONCLUSIONS: The findings of this investigation indicate that alcohol differentially affects aggressive responding in men and women. It is suggested that men are likely to express alcohol-related aggression in direct (shock intensity) and indirect (shock duration) forms, whereas women are most likely to use indirect forms (shock duration). PMID- 7475040 TI - College students' decisions to intervene in alcohol-related situations. AB - OBJECTIVE: In reviewing research on college students' intervention practices in alcohol-related situations, Seibold and Thomas recently advanced a transactional influence reconceptualization of intervention episodes. This study sought to explicate the factors contributing to college students' decision to intervene or not to intervene in alcohol-related situations. METHOD: In order to empirically explore this area, 489 students from three U.S. universities were surveyed about factors affecting their decisions to intervene or not in two types of alcohol related situations: a drunk-driving situation and an alcohol abuse situation. RESULTS: Students' intervention decisions were motivated primarily by concerns for the target and for potential consequences if they did not intervene. In contrast, decisions not to intervene were motivated primarily by an absence of a strong relationship with the target, their own condition at the time, and the potential impact of the intervention attempt on their image. Gender differences moderated many of these findings, however. CONCLUSIONS: Results are discussed in terms of approaching the study of alcohol interventions from a transactional influence perspective. PMID- 7475041 TI - Systemic and coronary hemodynamic actions and left ventricular functional effects of levosimendan in conscious dogs. AB - We examined the effects of levosimendan, a new myofilament Ca2+ sensitizer with phosphodiesterase (PDE)-inhibiting properties, on systemic and coronary hemodynamics and left ventricular (LV) systolic and diastolic function in conscious dogs with intact and blocked autonomic nervous system (ANS) reflexes. Twenty experiments were conducted in 10 dogs chronically instrumented for measurement of aortic and LV pressure, the peak rate of increase and decrease in LV pressure (+dP/dtmax and -dP/dtmin), subendocardial segment length, diastolic coronary blood flow (CBF) velocity, and cardiac output (CO). The slope (Mw) of the regional preload recruitable stroke work relation was used to assess myocardial contractility. Diastolic function was evaluated by -dP/dtmin, a time constant of isovolumic relaxation (tau), maximum segment lengthening velocity during rapid ventricular filling (dL/dtmax), and a regional chamber stiffness constant (Kp). Dogs were randomly assigned to receive levosimendan (0.5, 1.0, 2.0, and 4.0 micrograms.kg-1.min-1) with or without ANS blockade. On separate experimental days, systemic and coronary hemodynamics and LV pressure-segment length diagrams and waveforms were recorded after 10-min equilibration at each dose in the conscious ANS-intact or ANS-blocked state. Levosimendan increased heart rate (HR), CO, mean and diastolic CBF velocity, and pressure-work index (PWI, an estimate of myocardial oxygen consumption) and decreased LV end diastolic pressure (EDP), systemic vascular resistance (SVR), end-systolic and end-diastolic segment length, and mean and diastolic coronary vascular resistance (CVR) in dogs with intact ANS function. Levosimendan-induced increases in HR and PWI and decreases in SVR were attenuated by ANS blockade. Levosimendan caused equivalent dose-dependent increases in Mw in ANS-intact and ANS-blocked dogs, consistent with a positive inotropic effect independent of ANS activity. Levosimendan decreased tau (e.g., 35 +/- 1 ms during control to 29 +/- 1 ms at the high dose) and increased the magnitude of LV -dP/dtmin in dogs with intact but not blocked ANS reflexes, suggesting that relaxation was enhanced by favorable changes in systemic hemodynamics or ANS activation and direct effects of this drug on lusitropic state. Levosimendan also increased dL/dtmax to a greater degree in ANS-intact dogs, indicating that improvement of rapid ventricular filling was also partially dependent on ANS tone. No changes in Kp were observed in either experimental group. The results indicate that levosimendan decreases preload and afterload and has positive inotropic and lusitropic properties. The actions of levosimendan on diastolic function are largely mediated by the ANS. PMID- 7475042 TI - Effects of KRN4884, a novel K channel opener, on the cardiovascular system in anesthetized dogs: a comparison with levcromakalim, nilvadipine, and nifedipine. AB - Pharmacological profiles of KRN4884, 5-amino-N-[2-(2-chlorophenyl)ethyl]-N'-cyano 3-pyridinecarboxamidine+ ++, were evaluated in in vitro and in vivo experiments. In rat isolated aorta, KRN4884 (10(-10)-10(-5) M) produced a concentration dependent relaxation. Pretreatment with glibenclamide (10(-7)-10(-6) M) produced a rightward shift of the concentration-response curve for KRN4884. In anesthetized dogs, KRN4884 (3 and 10 micrograms/kg intravenously, i.v.), levcromakalim (3 and 10 micrograms/kg i.v.), nilvadipine (1-10 micrograms/kg i.v.), and nifedipine (1-10 micrograms/kg i.v.) produced decreases in mean blood pressure (MBP), total peripheral vascular resistance (TPR), and coronary vascular resistance (CVR), and increases in aortic blood flow (AoF) and coronary blood flow (CBF). The percentage decrease in CVR was greater than that in TPR with KRN4884 and levcromakalim, but nilvadipine and nifedipine showed no significant differences between CVR and TPR in percentage decreases. Heart rate (HR) was slightly increased by KRN4884 but was not affected by levcromakalim, nilvadipine, or nifedipine. Left ventricular dP/dt (LVdP/dt) was reduced only by nifedipine in a dose-dependent manner. The duration of the hypotensive action of KRN4884 was longer than those of levcromakalim and nifedipine and was similar to that of nilvadipine. The duration of the decreases in TPR and CVR induced by KRN4884 was longer than those induced by levcromakalim and nifedipine and shorter than that induced by nilvadipine. These results suggest that the cardiovascular effects of KRN4884 are very similar to those of the K channel opener levcromakalim and Ca channel blockers such as nilvadipine and nifedipine. However, the hypotensive effects of KRN4884 are long-acting in comparison with those of levcromakalim and the selective effect of KRN4884 on coronary vasculature is greater than those of nilvadipine and nifedipine. PMID- 7475043 TI - Effect of almokalant, a selective potassium channel blocker, on the termination and inducibility of paroxysmal supraventricular tachycardias: a study in patients with Wolff-Parkinson-White syndrome and atrioventricular nodal reentrant tachycardia. Almokalant PSVT Study Group. AB - Almokalant is a newly developed selective blocker of the delayed outward K+ current and exhibits the electrophysiological properties of a class III antiarrhythmic agent. In a Scandinavian multicenter, placebo-controlled trial, the antiarrhythmic efficacy of almokalant was investigated in patients with paroxysmal supraventricular tachycardia: 87 patients with mean age of 50 +/- 14 years (range 21-71 years), with reciprocating tachycardia due to either Wolff Parkinson-White (WPW) syndrome (n = 58) or atrioventricular nodal reentry tachycardia (AVNRT) (n = 29) were studied with transesophageal atrial stimulation. After a baseline procedure, during which sustained tachycardia was induced and overdrive terminated, tachycardia was reinduced and an intravenous (i.v.) infusion of either placebo or almokalant (aiming at a pseudoequilibrium plasma level of 20, 50, 100, or 150 nM) (Cpl 20-Cpl 150), was administered. Each patient was studied at two Cpl. Thirty-nine patients were randomly assigned in a double-blind fashion to either placebo+almokalant at Cpl 20 or Cpl 20 + Cpl 50; 26 patients were studied openly at Cpl 50 + Cpl 100, and 22 were studied openly at Cpl 100 + Cpl 150 almokalant. The antiarrhythmic efficacy was assessed as the ability to terminate induced tachycardia and to suppress inducibility: The proportion of patients in which the tachycardia was terminated was placebo 3 of 20 (15%); Cpl 20, 7 of 36 (19%): Cpl 50, 10 of 36 (28%); Cpl 100, 14 of 35 (40%); and Cpl 150, 5 of 9 (56%).(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7475044 TI - Progression and regression by verapamil of vitamin D3-induced calcific medial degeneration in coronary arteries of rats. AB - Vitamin D3-induced mural calcification represents an animal model for investigating experimental calcium (Ca) overload and calcification of arterial walls. In this study, long-term progression of calcific degeneration in coronary arteries of rats after one intoxication with vitamin D3 was examined, as well as possible regression of preestablished mural Ca overload with the Ca antagonist verapamil. Sprague-Dawley rats were treated with one intramuscular (i.m.) overdose of vitamin D3 [300,000 IU/kg body weight (b.w.)]. Oral verapamil therapy (100 mg/kg/day b.w. for 24 weeks) was initiated 14 days after the vitamin D3 intoxication. Arteriosclerotic alterations were verified by microchemical analyses of tissue Ca and of cholesterol contents with atomic absorption spectroscopy (special graphite tube technique) and gas chromatography, respectively, and by standard histological techniques. Serum lipids were determined by sequential ultracentrifugation. Between week 3 and week 26 after the vitamin D3 injection, a progressive Ca incorporation from 448.8 +/- 110 to 1,310 +/- 166.3% of control values (i.e., coronary Ca content in 32-week-old untreated control rats = 100%) was observed, associated with calcific morphological lesions, and reactive intimal plaque formation. Verapamil prevented this progression and induced a regression of preestablished mural Ca overload. Therefore, the coronary Ca content after 24 weeks of verapamil treatment amounted to only 146.3 +/- 53.8% of controls. The data indicate that an initial calcific lesion of coronary arteries may serve as crystallization nucleus for advancing Ca overload and morphological alterations.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7475045 TI - Effects of the angiotensin II receptor antagonist losartan on 24-hour blood pressure profiles of primary and secondary hypertensive rats. AB - Primary and secondary hypertension differ with regard to circadian blood pressure (BP) profiles. To evaluate the contribution of the renin-angiotensin system (RAS) to circadian BP regulation, we studied cardiovascular effects of the angiotensin II (AII) receptor antagonist losartan and the angiotensin-converting enzyme (ACE) inhibitor enalapril in animal models of primary and secondary hypertension after morning and evening dosing. Systolic/diastolic BP (SBP/DBP) and heart rate (HR) were measured telemetrically in spontaneously hypertensive rats (SHR) and transgenic hypertensive rats (TGR[mRen-2]27). Losartan (0.3 to 30 mg/kg) or enalapril maleate (10 mg/kg) were injected intraperitoneally (i.p.) either at 0700 or 1900 h. Baseline SBP/DBP and HR showed significant circadian rhythmicity in both strains. The 24-h means in SBP/DBP were 190/127 mm Hg in SHR and 200/139 mm Hg in TGR. TGR showed a reversed circadian profile in BP, with peaks occurring during the daily resting period, whereas HR peaked at night. Losartan reduced BP dose dependently; reductions in TGR were significantly greater and obtained at 30 fold lower doses than in SHR. Maximum decreases induced by losartan were similar to those induced with enalapril 10 mg/kg. Both drugs reduced BP in TGR more effectively when applied at 0700 than at 1900 h, resulting in a normalized circadian BP profile. Our results demonstrate that the RAS is involved in both the pathomechanism of hypertension and in the inverse circadian BP pressure pattern in TGR. PMID- 7475046 TI - Amiodarone is a dose-dependent noncompetitive and competitive inhibitor of T3 binding to thyroid hormone receptor subtype beta 1, whereas disopyramide, lignocaine, propafenone, metoprolol, dl-sotalol, and verapamil have no inhibitory effect. AB - The cardiovascular and electrophysiological effects of amiodarone resemble those of hypothyroidism. The drug has a structural resemblance to thyroid hormone (T3). Previous studies indicate that amiodarone exerts its major effect through antagonism of T3, probably as a result of inhibition of ligand binding to the thyroid hormone receptor (ThR). There are five subtypes of ThR, of which the beta 1 is the most prominent in the human heart. Our first aim was to investigate whether ThR is involved in a general antiarrhythmic mechanism for antiarrhythmic drugs or whether this action is specific for amiodarone. Therefore, we studied the affinity of one antiarrhythmic drug from every Vaughan-Williams group on T3 binding to human ThR beta 1 (hThR beta 1). Second, we wished to investigate whether amiodarone is a competitive or noncompetitive inhibitor. hThR beta 1, expressed in insect cells using a recombinant baculovirus, was used in regular binding competition assays. Disopyramide, lignocaine, propafenone, metoprolol, dl sotalol, and verapamil had no effect on T3 binding to hThR beta 1. Amiodarone showed a noncompetitive binding pattern at low concentrations (0.25-2 microM) and a competitive binding at high concentrations (2-8 microM). Among the antiarrhythmics tested, only amiodarone had affinity for hThR beta 1. This may represent a novel type of antiarrhythmic mechanism. The finding that amiodarone, in concentrations corresponding to therapeutic range in plasma, shifts from a noncompetitive to a competitive inhibitor, is of clinical interest in comparisons of low- and high-dose treatment. PMID- 7475047 TI - Effects of a new inotropic drug (DN-9693) on Ca2+ transients and contraction in ferret ventricular muscles. AB - We investigated the mechanisms of the effects of a newly synthesized cyclic AMP phosphodiesterase inhibitor (PDEI: 1,5-dihydro-7-(1-piperidinyl)imidazo[2,1 beta]quinazolin- 2(3H)-one dihydrochloride hydrate, DN-9693) on intracellular Ca2+ transients and tension in ferret papillary muscles. We used the aequorin method to measure Ca2+ transients in intact preparations. DN-9693 (1-100 microM) increased the peaks of both Ca2+ transients and tension. The potentiation of Ca2+ transients by DN-9693 was significant at lower concentrations (1-10 microM) but not at higher concentrations (50 and 100 microM). The peak of tension was significantly increased when the concentration of DN-9693 was increased (1-100 microM). In addition, the time to peak light was shortened at lower concentrations (1 and 5 microM). Other parameters of Ca2+ transients were not significantly altered. The time course of tension was not affected by DN-9693. In Triton X-100-treated skinned preparations, DN-9693 decreased the Ca2+ concentration required for half-maximal activation of the pCa-tension relation. These results suggest that the positive inotropic effect of DN-9693 is attributable to the alteration in Ca2+ handling of sarcoplasmic reticulum (SR) and Ca2+ sensitization of the myofilaments. PMID- 7475048 TI - Effects of losartan on angiotensin and bradykinin peptides and angiotensin converting enzyme. AB - Antagonists of the type 1 (AT1) angiotensin II (Ang II) receptor increase renin secretion and plasma Ang II levels, and the increased Ang II levels may counteract the effects of the antagonist. Moreover, other investigators have suggested that the reactive increase in Ang II levels may increase bradykinin (BK) levels through stimulation of the type 2 Ang II receptor (AT2). We investigated the acute effects of the AT1 receptor antagonist losartan (intraarterial injection of 10 mg/kg every 12 h) in male Sprague Dawley rats by measuring circulating angiotensin and BK peptides at 6, 12, and 24 h. Whereas acute losartan administration increased blood angiotensin levels four- to sixfold, blood BK levels were unchanged. We also investigated the effects of losartan administered for 8 days (10 mg/kg every 12 hours, by intraperitoneal injection) on circulating and tissue levels of angiotensin and BK peptides, and angiotensin-converting enzyme (ACE). Losartan increased plasma renin levels 100 fold; plasma angiotensinogen levels decreased to 24% of control; and plasma aldosterone levels were unchanged. Ang II levels in plasma, adrenal, lung, heart, and aorta were increased 25-, 8-, 3.5-, 2.4-, and 14-fold, respectively, by losartan administration. By contrast, kidney Ang II levels decreased to 71% of control, accompanied by a decrease in kidney levels of BK-(1-7) and BK-(1-9). No other tissue showed a change in BK peptide levels, except for a reduction in blood levels of BK-(1-8) to 43% of control. Plasma ACE increased by 13-50%, but tissue ACE levels were unchanged. These data demonstrate that losartan has tissue specific effects on endogenous levels of angiotensin and BK peptides and indicate that increased BK levels do not contribute to the actions of losartan. The absence of a reactive increase in endogenous kidney levels of Ang II indicates that this tissue is likely to be the most sensitive to AT1 receptor antagonism. PMID- 7475050 TI - Effects of acetaldehyde on the isolated papillary muscle of diabetic rats. AB - The effects of acetaldehyde (ACA) were examined in isolated electrically driven papillary muscle preparations from normal and streptozotocin-treated diabetic rats. Muscles from diabetic rats developed greater tension than those from normal rats. In muscles from both groups, ACA caused concentration-dependent negative inotropic effects that were independent of cholinergic or purinergic mechanisms and were not attributable to nitric oxide (NO) release. ACA was three to five times more potent with regard to its negative inotropic effect in diabetic than in normal rat muscles. A propranolol-sensitive, sympathetically mediated positive inotropic effect occurred at certain concentrations. Decreasing [Ca2+]o from 2.7 to 0.5 mM reduced basal developed force to a significantly greater extent in muscles from normal rats than in those from diabetic rats. In low [Ca2+]o, concentration-response curves to CaCl2 in diabetic muscles were displaced to the left of that in normal muscles, suggesting that diabetic muscles are more sensitive to the positive inotropic effect of added CaCl2 at low [Ca2+]o, whereas at higher [Ca2+]o (> 1 mM), normal muscles developed more force in response to added CaCl2. ACA 10 and 30 mM more readily inhibited CaCl2-induced positive inotropic effect in normal than in diabetic muscles. Force-frequency curves, (negative staircase response) were recorded in both normal and diabetic muscles. In diabetic muscles, the curve exhibited a positive component at the lowest frequencies applied and was displaced to the right of that in normal muscle. ACA concentration-dependently inhibited force development, and diabetic muscles were more susceptible to the negative inotropic effect of ACA, when the stimulation frequency was increased.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7475049 TI - Monoclonal antibody [7E3 F(ab')2] prevents arterial but not venous rethrombosis. AB - The role of the platelet glycoprotein IIb/IIIa (GPIIb/IIIa) receptor and the efficacy of GPIIb/IIIa receptor antagonists on reocclusion after arterial versus venous thrombolysis is unknown. We used a canine model of simultaneous carotid artery and jugular vein thrombosis to evaluate the effect of the murine monoclonal antibody 7E3 [7E3 F(ab')2] on intravascular thrombotic reocclusion after thrombolysis in both vessels. The left carotid artery and right jugular vein were instrumented with flow probes, a critical stenosis, and an electrode through which an anodal current was applied to induce formation of an occlusive thrombus. After persistent occlusion of both vessels, 7E3 (0.8 mg/kg, n = 7) or saline (n = 10) was administered intravenously (i.v.) immediately after the local administration of anisolylated plasminogen streptokinase activator complex (APSAC, 0.1 U/kg). Ex vivo platelet aggregation in response to ADP or arachidonic acid was inhibited completely, and bleeding time was increased threefold by 7E3. The administration of 7E3 did not affect the activated partial thromboplastin time as compared with control values. The time to reperfusion of either vessel was not altered significantly in the presence of 7E3. Arterial or venous thrombolysis after APSAC was achieved in 10 of 10 and 8 of 10 control animals, respectively, but rethrombosis occurred in both the carotid artery and jugular vein in each of the saline-treated animals. Treatment with 7E3 prevented cyclic flow variations and reocclusion in the carotid artery (p < 0.05) in all treated animals (n = 7). In contrast, 7E3 did not suppress cyclic flow variations in the jugular vein, and rethrombosis occurred in five of six vessels despite the presence of platelet inhibition as assessed ex vivo. Residual arterial thrombus weights were reduced by pretreatment with 7E3 (saline = 24 +/- 4 mg, 7E3 = 11 +/- 3 mg; p < 0.05), whereas residual venous thrombus weights were not affected (saline 25 +/- 5 mg and 7E3 26 +/- 11 mg). The results indicate that inhibition of the platelet GPIIb/IIIa receptor does not prevent jugular vein rethrombosis despite its ability to prevent rethrombosis in the carotid artery. Different thrombotic mechanisms are involved in forming arterial and venous thrombi. Interventions that modulate arterial thrombotic events need not affect occlusive thrombotic activity in the venous circulation. PMID- 7475051 TI - Use-dependent effects of the class III antiarrhythmic agent NE-10064 (azimilide) on cardiac repolarization: block of delayed rectifier potassium and L-type calcium currents. AB - We studied the effects of NE-10064 (azimilide), a new antiarrhythmic agent reported to be a selective blocker of the slowly activating component of the delayed rectifier, IKs. In ferret papillary muscles, NE-10064 increased effective refractory period (ERP) and decreased isometric twitch tension in a concentration dependent manner (0.3-30 microM). Increases in ERP showed reverse use-dependence, and were greater at 1 than at 3 Hz. In contrast, changes in tension were use dependent, with larger decreases observed at 3 than at 1 Hz. In guinea pig ventricular myocytes, NE-10064 (0.3-3 microM) significantly prolonged action potential duration (APD) at 1 Hz. At 3 Hz, NE-10064 (0.3-1 microM) increased APD only slightly, and at 10 microM decreased APD and the plateau potential. NE-10064 potently blocked the rapidly activating component of the delayed rectifier, IKr (IC50 0.4 microM), and inhibited IKs (IC50 3 microM) with nearly 10-fold less potency. NE-10064 (10 microM) did not block the inward rectifier potassium current (IKl). NE-10064 (10 microM) blocked the L-type calcium current (ICa) in a use-dependent manner; block was greater at 3 than at 1 Hz. We conclude that (a) NE-10064's block of potassium currents is relatively selective for IKr over IKs, (b) NE-10064 inhibits ICa in a use-dependent fashion, and (c) NE-10064's effects on ERP and tension in papillary muscle as well as APD and action potential plateau level in myocytes may be explained by its potassium and calcium channel blocking properties. PMID- 7475052 TI - Effect of nitric oxide donors on neointima formation and vascular reactivity in the collared carotid artery of rabbits. AB - Intimal thickening in arteries is considered a site of predilection for atherosclerosis. We investigated whether oral application of the nitric oxide (NO) donors SPM-5185 [N-nitratopivaloyl-S-(N'-acetylalanyl)-cysteine ethylester, 10 mg/kg body weight twice daily (b.i.d.)] and molsidomine (10 mg/kg body weight/day) can retard neointima formation and changes in vascular reactivity induced by a nonocclusive, soft silicone collar positioned around the left carotid artery of rabbits. The contralateral carotid artery was sham operated and served as a control. Drug and placebo (diet without drug) treatments were initiated 7 days before placement of the collar. At the end of the experiments, two segments were cut from each collared and sham-treated artery, one for measurement of the cross-sectional area of intima and media and the other for isometric tension recording. Sham treatment did not result in intimal thickening in either group. In contrast, the intima/media (I/M) ratio was considerably increased after 14 days of collar treatment as a result of neointima formation. Intimal thickening was significantly inhibited by SPM-5185 (I/M ratio 0.05 +/- 0.01 vs. 0.11 +/- 0.02, p < 0.05), but not by molsidomine (0.06 +/- 0.02 vs. 0.08 +/- 0.02, p = 0.49), which is a donor of both NO and superoxide anions. Neither collar nor NO donor treatment altered the area of the media. SPM-5185 did not alter the percentage of replicating smooth muscle cells (SMC) in the media after collar treatment, as demonstrated by their immunoreactivity for proliferating cell nuclear antigen (PCNA).(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7475053 TI - Pharmacological analysis in rat of the role of the ATP-sensitive potassium channel as a potential target for antifibrillatory intervention in acute myocardial ischaemia. AB - We tested the hypothesis that blockade of the ATP-sensitive K+ channel (IK(ATP)) is an antiarrhythmic mechanism in acute myocardial ischaemia, using an opener of the channel (10 microM RP 49356, RP) and a blocker of the channel (10 microM glibenclamide, GL) and a combination of the two drugs (GL+RP, 10 microM each) in a randomised blinded study. Isolated rat hearts (n = 8 per group) were subjected to 30-min left regional ischaemia. GL and GL+RP widened QT interval after 10-min ischaemia (197 +/- 39 and 203 +/- 20 ms, respectively vs. 154 +/- 12 ms in controls), whereas RP significantly shortened QT interval (123 +/- 6 ms). GL and GL+RP decreased coronary flow (p < 0.05). RP caused slight increase in flow during ischaemia. These effects are all consistent with modulation of vascular and cardiac IK(ATP). RP alone had no effect on ischaemia-induced arrhythmias. Neither did GL have any effect on the incidence of ventricular fibrillation (VF: 88 vs. 100% in controls). However, GL reduced the incidence of sustained VF (VF lasting continuously for > 2 min) to 14% vs. 88% in controls (p < 0.05). Therefore, GL had defibrillatory activity. Surprisingly, in view of these findings, the GL+RP combination significantly reduced the incidence of VF to 25% (from 100% in control hearts, p < 0.05) i.e., had an antifibrillatory effect. So, two agents that produce pharmacological effects attributable to block and opening of IK(ATP) when administered singly had no effects on the incidence of ischaemia induced VF.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7475054 TI - Pharmacologic profile of the dihydropyrimidine calcium channel blockers SQ 32,547 and SQ 32,926 [correction of SQ 32,946]. AB - SQ 32,926 and SQ 32,547, two dihydropyrimidine calcium channel blockers, were characterized as potent inhibitors of depolarization-induced contractions of isolated smooth muscle preparations. In rat aorta, the IC50 values were 5.5 nM for SQ 32,547 and 8.1 nM for SQ 32,926, values which compare favorably with that of 2.9 nM for nifedipine. The dihydropyrimidines were also tested in a model of stable angina: pacing-induced ischemia in dogs. Both SQ 32,547 and SQ 32,926 reduced the ST-segment elevation observed in vehicle-treated animals. No significant changes in hemodynamic status were detected, suggesting that a reduction in cardiac work secondary to afterload reduction was probably not a major contributor to the protective effects. Neither was increased coronary blood flow important for the antiischemic outcome because no significant effects of the dihydropyrimidines were observed on ischemic regional blood flow. SQ 32,547 was also studied in globally ischemic, isolated rat hearts. In this model, SQ 32,547 was protective because it significantly reduced contracture formation and lactate dehydrogenase (LDH) release. Washing out the effect of SQ 32,547 in isolated hearts and smooth muscles was difficult, presumably due to its lipophilicity. In the smooth muscle preparations, the effects of both nifedipine and SQ 32,926 were much more easily washed out. As with other calcium channel blockers, increasing the antiischemic effects of SQ 32,547 was associated with a higher cost in terms of cardiac function. In summary, the two novel dihydropyrimidines, SQ 32,547 and SQ 32,926 showed antiischemic properties in animal models. PMID- 7475055 TI - Cardiovascular effects of the novel potassium channel opener UR-8225. AB - We compared the hemodynamic and electrophysiological effects of UR-8225, a new potassium channel opener with those of levcromakalim. UR-8225 and levcromakalim (0.03-1 mg/kg) dose-dependently decreased mean arterial pressure (MAP) in conscious spontaneously hypertensive rats (SHR) and deoxycorticosterone-salt (DOCA) hypertensive rats and in conscious and anesthetized normotensive rats. The decrease in MAP was accompanied by a dose-dependent increase in heart rate (HR). Levcromakalim was about twice as potent as an antihypertensive agent, and its hypotensive and tachycardic effects were of longer duration than those of UR 8225. In conscious normotensive rats, the hypotensive response to UR-8225 (1 mg/kg) did not diminish with repeated dosing for 16 days. In conscious SHR, propranolol antagonized the tachycardic response of UR-8225 without affecting its hypotensive response. In anesthetized rats, glibenclamide (3, 10, and 20 mg/kg) dose-dependently reduced the hypotensive effect of the drug. In anesthetized dogs, UR-8225 and levcromakalim (0.01-30 micrograms/kg) dose-dependently decreased MAP and total peripheral resistance (TPR): UR-8225 was approximately 10 times less potent than potent than levcromakalim. UR-8225 had no effect on HR, left ventricular end-diastolic pressure (LVEDP), stroke volume (SV), or cardiac output (CO) but decreased rate-pressure product (RPP) and cardiac work (CW). In contrast, levcromakalim decreased HR, RPP, and CW and increased SV and LVEDP, whereas CO remained unaltered. UR-8225 and levcromakalim (0.1-10 microM) had no effect on the ventricular action potentials (APs), whereas at higher concentrations they shortened the AP duration (APD), an effect that was antagonized by glibenclamide. UR-8225 and levcromakalim decreased the Vmax and shortened the APD of the slow ventricular APs elicited by isoprenaline in K(+) depolarized ventricular muscle fibers. These results indicated that UR-8225 decreases blood pressure (BP) by reducing TPR. Its electrophysiological and hemodynamic effects were blocked by glibenclamide, which suggests that they may be mediated largely through the activation of ATP-sensitive K+ channels. PMID- 7475056 TI - Insulin sensitivity in obese hypertensive dyslipidemic patients treated with enalapril or atenolol. AB - We evaluated the effects of enalapril [angiotensin converting enzyme (ACE) inhibitor] in comparison with atenolol (beta-blocker) on insulin sensitivity and serum lipoprotein concentration in obese hypertensive dyslipidemic patients. Twenty-eight hypertensive [mean blood pressure (MAP) 152 +/- 3/103 +/- 1 mm Hgl], obese [mean body mass index (BMI) 30 + 1 kg/m2A], dyslipidemic [total triglycerides 2.0 +/- 0.2 mM and/or high density lipoprotein (HDL) cholesterol 1.1 +/- 0.1 mM and low density lipoprotein (LDL) cholesterol 4.5 +/- 0.2 mM] outpatients were randomized in two groups receiving enalapril or atenolol for 12 weeks, in an investigator-blinded, parallel, comparative two-center trial. Insulin sensitivity was assessed by a modified insulin suppression test. Blood pressure (BP), insulin sensitivity, and serum lipoprotein concentrations were compared before and after each treatment and between the two treated groups. BP decreased significantly and comparably during enalapril and atenolol treatment (p < or = 0.01). The sensitivity to insulin improved by 15% (p = 0.03) in the enalapril group and worsened by 17% (p < or = 0.01) in the atenolol group. Serum lipoprotein concentrations were not modified by any treatment. The improvement in insulin sensitivity caused by enalapril treatment appears to be an advantage as compared with atenolol treatment in hypertensive obese and dyslipidemic patients, whereas the BP-lowering efficacy of the two drugs is similar. Because this effect has been reported with other ACE inhibitors, it appears to be characteristic of the entire class of ACE inhibitors. PMID- 7475057 TI - Differential effects of the calcium antagonist mibefradil in epicardial and intramyocardial coronary arteries. AB - The efficacy of vasodilation depends on the drug and the contractile agonist involved. Furthermore, vasodilation also may be detrimental (i.e., coronary steal), possibly depending on the anatomic site and/or action of the vasodilator. We investigated the effects of Ca2+ antagonism in porcine epicardial and intramyocardial coronary arteries suspended in organ chambers filled with physiological salt solution (95% O2/5% CO2, 37 degrees C); isometric tension was measured. In epicardial vessels contracted with KCl, the thromboxane analogue U 46619, or endothelin-1 (ET-1), relaxations to the Ca2+ antagonist mibefradil were most effective after precontraction with KCl, followed by U 46619 and ET-1. In intramyocardial vessels, the contraction to KCl, U 46619, and particularly ET-1, was much more effectively inhibited by mibefradil than in epicardial arteries. In vessels preincubated with mibefradil, the drug was moderately effective in preventing the initiation of contractions. Preincubation with the inhibitor of nitric oxide production (NO), L omega-nitro arginine methyl ester (L-NAME) or endothelium removal, did not increase relaxations to mibefradil. However, epicardial but not intramyocardial vessels incubated with mibefradil exhibited enhanced relaxations to bradykinin (BK) and sodium nitroprusside (SNP) as compared with control. Thus, Ca2+ antagonism is particularly effective in intramyocardial coronary arteries. In addition to its inhibitory effects on contractility, calcium antagonism facilitates the effects of endothelium-derived NO in epicardial coronaries at the level of vascular smooth muscle (VSM). PMID- 7475058 TI - Electrophysiological and arrhythmogenic effects of the histamine type 1-receptor antagonist astemizole on rabbit Purkinje fibers: clinical relevance. AB - Astemizole is a potent histamine H1-antagonist that has been associated with cases of life-threatening cardiac arrhythmias, including torsade de pointes and atrioventricular (AV) block. However, its effects on cardiac action potential (AP) has not been described. We examined the electrophysiological effects of astemizole on rabbit Purkinje fibers using conventional glass microelectrodes in parallel with the effects of the widely used histamine H2-antagonist cimetidine, selected because it has no known cardiac arrhythmic toxicity. Astemizole (0.01-3 microM) exerted a concentration-dependent prolonging effect on final repolarization that did not reach steady state after 3 h of exposure. This effect was more pronounced at low stimulation frequency and was less marked at high stimulation frequency. In addition, early afterdepolarizations (EADs) occurred in one third of the fibers. Increasing extracellular concentration of KCl (2.7-5.4 mM) or MgCl2 (1-5 mM) suppressed EADs and reversed the prolonging effect that was conversely exaggerated by decreasing KCl (4-2.7 mM) or MgCl2 (1-0.5 mM) concentration. At higher concentrations (3-30 microM), astemizole induced an increasing depressant effect on the maximal rate of depolarization (Vmax) that became more pronounced with high stimulation frequency. All parameters were strongly depressed at 10 microM astemizole, leading to cellular inexcitability in 5 of 12 fibers when exposed to 30 microM astemizole. In comparison, cimetidine induced minor changes on AP characteristics, i.e., a prolongation in plateau duration at high (30-100 microM) concentrations. These results provide evidence that astemizole exerts quinidine-like effects on cardiac APs that are compatible with the occurrence of the clinically observed arrhythmias. PMID- 7475059 TI - Depressed sodium-calcium exchange in human umbilical arteries from women with preeclampsia. AB - Responses to low-sodium solution were studied in helical preparations of human umbilical arteries from women with preeclampsia and age-matched normotensive women. Reduction of extracellular sodium concentration (replaced by N-methyl-D glucamine, NMG) and consequent restoration of extracellular sodium-evoked contractions and relaxations, respectively, in the arteries pretreated with phenoxybenzamine, caffeine, ryanodine, and verapamil. The rate and magnitude of the contraction induced by low-sodium solution and the rate of the relaxation induced by the restoration were less in preeclampsia than in normotension. Therefore, the activity of Na-Ca exchange is depressed in human umbilical arterial smooth muscle in preeclampsia, suggesting a possibility that the depressed activity leads to an abnormal regulation of arterial tone in preeclampsia. PMID- 7475060 TI - Relaxant responses to prostaglandin F2 alpha and E2 of isolated human uterine arteries. AB - We wished to determine the action of prostaglandins (PG) and to analyze pharmacologically the mechanisms of their action in isolated human uterine arteries in special reference to mediators liberated from the endothelium and subendothelial tissues. Helical strips of the human uterine artery with and without the endothelium were suspended in the Ringer-Locke solution for isometric tension recording. The relaxant response to PGF2 alpha was reversed to a contraction by cyclooxygenase inhibitors and suppressed by tranylcypromine, a PGI2 synthase inhibitor, but was not influenced by endothelium denudation. Relaxations induced by PGE2 and beraprost, a PGI2 analogue, were augmented by cyclooxygenase inhibitors and tranylcypromine but were not affected by ONO3708, an antagonist of vasoconstrictor prostanoids, and endothelium denudation. The potentiating effect of indomethacin was observed in the strips both with and without the endothelium and was antagonized by treatment with beraprost. The relaxation caused by PGF2 alpha apparently is mediated by PGI2 released from subendothelial tissues, whereas the PGE2-induced relaxation is due to the direct action on smooth muscle; the action may be eliminated by the basal release of PGI2 from subendothelial tissues. PMID- 7475061 TI - Effect of nitric oxide synthase inhibition on bleeding time in humans. AB - Endothelium-derived nitric oxide (NO) is a potent in vitro inhibitor of platelet adhesion and aggregation, but its role in regulating platelet reactivity in vivo and in humans in particular is undefined. Our primary objective was to determine whether the in vivo inhibition of NO production shortens template bleeding time (BT). The hemodynamic and platelet effects of NG-mono-methyl-L-arginine (L-NMMA), an NO synthase inhibitor, were studied in 12 normal volunteers. Forearm template BT was determined before and 15 min after the intravenous (i.v.) infusion of 4.3 mg/kg L-NMMA. L-NMMA infusion increased mean arterial pressure from 88 +/- 4 to 99 +/- 3 mm Hg (p = 0.0001). Plasma NO levels, determined by chemiluminescence, decreased 65 +/- 10% from basal values (p < 0.05), confirming inhibition of endogenous NO production. Intravenous L-NMMA shortened BT from 5.5 +/- 0.9 to 4.0 +/- 0.6 min (p = 0.026), or by 24 +/- 8% (p = 0.008). L-NMMA infusion did not significantly change ex vivo platelet aggregation. To determine whether vasoconstriction affected BT, we investigated forearm blood flow (FBF; determined by venous occlusion plethymography), and template BT in 3 subjects after the local infusion of L-NMMA (2-4 mg/min intraarterially, i.a.). Intraarterial administration of L-NMMA caused a 39 +/- 3% (p = 0.006) reduction in FBF in the infused arm but did not change BT. These data show that systemic inhibition of NO production shortens BT in humans. However, the precise mechanism by which L-NMMA shortens BT is not completely defined. PMID- 7475062 TI - Triad for change--a new concept for cancer prevention, control, treatment, and rehabilitation. PMID- 7475063 TI - Analysis of proliferative activity using antiproliferating cell nuclear antigen antibody in colorectal cancer. AB - Using anti-PCNA antibody (PC10), an immunohistochemical study of the expression of PCNA in formalin-fixed and paraffin-embedded materials of colorectal cancer patients was performed and correlation of PCNA expression with clinicopathological findings and DNA ploidy pattern was studied. PCNA labeling rate (PCNA LR) was estimated in the advancing margin of the tumor and ranged from 23.8% to 77.9%. There was a significant difference in lymphatic vessel invasion, liver metastasis and Dukes' stage between the groups with high (> 48.7) and low (< 48.7) PCNA LR (P < 0.05). No differences were seen in tumor size, histological type, lymph node metastasis or DNA ploidy pattern. In patients with younger age, infiltration to neighboring organs, a high degree of venous invasion, and peritoneal dissemination, the frequency of high PCNA LR tended to be higher (P < 0.1). The results above suggest that a high proliferative activity as defined by evaluation of the PCNA LR at the advancing margin of the tumor may be one of the parameters of malignant potential and helpful as a predictor of liver metastasis in colorectal cancer. PMID- 7475064 TI - Effects of intraportal administration of chemoimmunotherapeutic agents on natural killer cell activity in the rat liver. AB - To evaluate the effects of continuous intraportal chemotherapy on tumor immunity of the liver, natural killer (NK) cell activity was estimated in the rat liver. In an in vitro study, NK-enriched mononuclear cells collected from rat liver were incubated with 5-fluorouracil (5-FU) alone and with a 5-fluorouracil/mitomycin C (5-FU/MMC) combination at different concentrations. NK activity was measured after 24-hour incubation. In an in vivo study, the continuous intraportal administration of chemotherapeutic (5-FU/MMC) and chemoimmunotherapeutic (5 FU/MMC/lentinan) agents was carried out in rats for 5 days, and NK-enriched mononuclear cells were then collected from the liver for the measurement of NK activity. Neither 5-FU alone nor the 5-FU/MMC combination affected NK activity in vitro. In the in vivo study, however, the 5-FU/MMC combination significantly decreased NK activity, and the addition of lentinan recovered the activity to the control level. It can thus be concluded that the intraportal administration of chemotherapeutic agents reduces NK activity in the liver and the addition of an immunostimulator to such agents prevents this reduction. PMID- 7475065 TI - Argyrophilic nucleolar organizer region (AgNOR) area per nucleus as a prognostic factor in breast cancer. AB - Whether argyrophilic nucleolar organizer region (AgNOR) is a predictor of prognosis in breast cancer is controversial. The semiquantitative procedure used to calculate interphasic argyrophilic dots optically is neither reliable nor reproducible. We measured the mean area of AgNOR dots, the mean AgNOR area per nucleus (MA), and the mean AgNOR number per nucleus (MN) by automated image analysis in specimens from 131 patients with breast cancer. Higher MA (> 7.41 microns2), detected in 22 (16.8%) of 131 patients, did not correlate with clinicobiologic variables, except for DNA ploidy status. Overall survival was significantly better in patients with lower MA than in those with higher MA. On multivariate analysis, MA was a significant independent factor, followed by nodal status. These findings indicate that expression of AgNOR should be evaluated in terms of area occupied and that the mean area of AgNOR per nucleus (MA) is an important prognostic factor in the overall survival of breast cancer patients. PMID- 7475066 TI - Mastectomy as the preferred treatment for breast cancer among new immigrants from the former USSR. AB - Sixty-four new immigrant patients with primary breast cancer who arrived in Israel since 1990 were compared to 322 Israeli women with primary breast cancer. New immigrant patients had a more advanced stage at diagnosis (stage III+IV 28% vs. 13% respectively, P = 0.00005); larger mean tumor size (3.08 +/- 1.7 cm vs. 2.25 +/- 1.4 cm respectively, P = 0.00002); and more involved lymph nodes (3.8 +/ 5.3 vs. 1.8 +/- 3.3 P = 0.0002). Forty-seven percent of new immigrants had breast-conserving surgery compared to 69% of the Israelis (P = 0.0004). In stage II 51% of the new immigrants had conservative surgery compared to 74% in the Israeli group (P = 0.03). Mastectomy was recommended to 39 Israeli patients and to six immigrants. Only 19% of the Israeli group requested mastectomy compared to 44% in the new immigrant group. Most of the new immigrant patients chose mastectomy although breast-conserving surgery was an equal alternative. The reasons for choosing mastectomy as the preferred mode of treatment were related to economic problems, different life concepts, and other priorities. Constant surveillance and considerable enlightenment are needed in the new immigrant population, so that they will be able to separate the disease and its treatment from their paramedical problems. PMID- 7475067 TI - Follow-up of patients resected for gastric cancer. AB - In this study we used a cost-outcome analysis to evaluate our follow-up protocol for patients who had been resected for gastric cancer. We designed a descriptive cross-sectional trial through consecutive sampling of patients who had undergone resection of gastric carcinoma and were followed in our outpatient department during 1991. Serological (CEA) and or imaging procedures were pathologic at least two months prior to the onset of symptoms in 33% of recurrences. No significant correlation was found between serum CEA levels and CEA tumor tissue staining in patients who recurred. Only 17% of patients who relapsed underwent further treatment (surgery and chemotherapy) with no improvement found in terms of survival. The overall cost per year has been estimated at US$ 6118. Our results show that serological levels of CEA and available imaging techniques for routine follow-up provide little advantage in diagnosing gastric cancer recurrence over clinical surveillance alone. PMID- 7475068 TI - The combination 5-fluorouracil/levamisole induces enhanced rat Kupffer cell mediated cytotoxicity in vitro against the syngeneic colon adenocarcinoma cell line CC531. AB - The mode of action of the combination treatment 5-fluorouracil (5-FU) and levamisole in colorectal cancer patients is unknown. It is postulated that the beneficial effect may be explained by an immunomodulatory effect on Kupffer cell (KC) cytotoxicity. We evaluated the effect of levamisole (200 micrograms/ml) and 5-FU (10 microM) on rat KC cytotoxicity against syngeneic CC531 tumor cells. Viability of KCs was unaffected by 5-FU and/or levamisole. The combination did not enhance growth inhibition of CC531 compared to 5-FU alone. A significant increase in KC cytotoxicity was observed after 24-hr incubation with 5 FU/levamisole especially at an effector/target ratio of 10 (P < 0.05). 5-FU alone had no effect on KC cytotoxicity, while levamisole induced only a slight increase. Our in vitro data suggest that the additive effect of the combination 5 FU/levamisole on KC cytotoxicity may attribute to the beneficial effect of the adjuvant treatment in colorectal cancer patients. PMID- 7475069 TI - Intraoperative histologic assessment of surgical margins and lymph node metastasis in breast-conserving surgery. AB - The diagnostic value of intraoperative histologic examination of frozen sections of surgical margins and axillary lymph nodes (AX) was investigated in 95 patients with breast cancer who underwent breast-conserving surgery. The periphery of the excised breast tissue was peeled like an orange and examined histologically by frozen section. The results were compared with examination by permanent section. Evaluation of surgical margins by frozen section resulted in a diagnostic accuracy of 87%, a sensitivity of 96%, and a specificity of 84%. Enlarged or hardened AXs were sampled from the axillary pad which was derived from a complete AX dissection. Histologic examination using frozen section was performed during surgery. After the operation, the remaining AXs were removed from the axillary pad by hand dissection and histologically examined on permanent section. A diagnostic accuracy of 97%, a sensitivity of 77%, and a specificity of 100% were achieved in the diagnosis of AX involvement on frozen section. It was therefore concluded that intraoperative histologic examination of frozen sections may be useful in the determination of involvement of the surgical margins and the AXs in patients with breast cancer. PMID- 7475070 TI - Long-term follow-up of patients with superficial transitional cell carcinoma of the urinary bladder treated by intravesical mitomycin C. AB - Prophylactic intravesical chemotherapy (IVC) reduces recurrence rates of superficial transitional cell carcinoma (TCC) of the urinary bladder. The patient cohort existed of 86 individuals (stage TaN0M0 or T1N0M0) superficial carcinoma of various grades of malignancy. Following initial transuretheral resection or diagnostic cystoscopy, mitomycin C (MMC), 20 mg dissolved in 50 ml saline, was instilled intravesically by catheter over 1 hr at 2-week intervals, initially and then four more times followed by diagnostic cystoscopy (one course = 12 weeks). Two similar courses were administered thereafter for a total period of 36 weeks. For patients in remission, installations continued in monthly fractionations for 9 more months (cystoscopy every 3 months) and then at 2-month intervals for 12 more months (cystoscopy every 6 months). When cystoscopy revealed recurrence, IVC was repeated from the beginning. No symptoms of MMC-related toxicity were observed. Cystoscopic follow-up evaluations showed a complete response rate of 84% at 3 years and 81% at 5 years after initial therapy. Twenty-seven patients who had not responded to previous treatment with other drugs responded to MMC. PMID- 7475071 TI - Venous gas embolism during cryosurgery for bone tumors. AB - Cryosurgery using liquid nitrogen is a method for treating benign- and low malignant skeletal tumors. The advantage of preserving the supportive function of bone should be compared to the risk for its complications; postoperative fracture is well known, but less so the occurrence of intraoperative venous gas embolism. This paper describes 17 patients: 2 patients who had serious hemodynamic complications during cryosurgery and a study of 15 patients in whom end-tidal N2 tension was measured in an attempt to investigate the clinical incidence of venous gas embolism during cryosurgery. In the 15 cases analyzed, we did not detect any exhaled N2 during cryosurgery. PMID- 7475072 TI - Absorption cytometric DNA analysis of sections of gastric carcinomas and a comparison with cytofluorometry of single-cell suspensions and flow cytometry. AB - Measurements were made of cellular deoxyribonucleic acid (DNA) content in paraffin-embedded materials from 44 gastric carcinomas, using three different methods; static cytometric light absorbance technique using tissue sections (SCM abs), static cytometric fluorescence technique using single-cell suspensions (SCM flu), and flow cytometry (FCM). The DNA ploidy determined by SCM-abs was grouped into low or high ploidy according to dispersion on the DNA histogram, and the DNA ploidy determined by SCM-flu and FCM was grouped into diploidy or aneuploidy, according to the peak DNA value. Accord between SCM-abs and SCM-flu or between SCM-abs and FCM was confirmed when low ploidy by SCM-abs showed diploidy by SCM flu or FCM and high ploidy by SCM-abs showed aneuploidy by SCM-flu or FCM. An incomplete but good accord of DNA ploidy was obtained with these methods. PMID- 7475073 TI - Substernal goiter. AB - The management of a substernal goiter is a problem which has challenged surgeons since its first description in 1749. While the overall incidence in the United States has decreased with the routine use of iodized salt, the development of large multinodular substernal goiters in the rest of the world is still common. In addition, even in those regions where they are less common, knowledge of their treatment is important as they can represent up to 7% of mediastinal tumors. Certainly, the majority are large, benign masses found in the superior and anterior mediastinum, although from 3 to 15% can be malignant in nature. The presenting symptoms generally relate to the compressive nature of the mass on nearby structures. Up to 90% of patients report some form of respiratory symptoms in association with these masses. Diagnostic evaluation should include chest x ray and computed tomographic (CT) scan. Needle aspiration biopsy should be avoided due to its dangerous substernal location. The treatment is surgical, as medical therapy is generally unsuccessful. Perioperative management should include careful evaluation of the airway as the extent of compression and deviation caused by the mass can lead to a difficult intubation. The vast majority of substernal goiters can be removed via a cervical incision; occasionally sternotomy or thoracotomy is necessary. Although rare, tracheomalacia secondary to prolonged compression of the trachea by the mass needs to be watched for postoperatively. Overall, the results of surgical treatment are excellent, as morbidity and mortality are minimal and patients can expect full relief of symptoms secondary to these mediastinal masses. PMID- 7475074 TI - Familial adenomatous polyposis and lung cancer. AB - A 43-year-old man presented with a lung nodule 19 years after undergoing a total colectomy for familial adenomatous polyposis (FAP). There had been no evidence of malignant transformation in the colectomy specimen, and current gastrointestinal investigation did not reveal evidence of tumor. Pathological analysis of the lung nodule demonstrated adenocarcinoma of the lung of the fetal type. This is the first reported case of a lung neoplasm in a patient with FAP. The development of an unusual lung tumor in a patient with FAP, a condition associated with other extracolonic tumors, suggests that there may be an association between the two conditions. PMID- 7475075 TI - Technical details facilitating resection of large numbers of tumors metastatic to lungs. PMID- 7475076 TI - The role of the ocular lens in the regulation of intraocular pressure. AB - The relationship between lenticular colloid osmotic (oncotic) pressure and intraocular pressure is discussed, with particular reference to a postulated buffering reservoir action of the lens in protecting against acute changes in intraocular pressure. A hypothesis is developed regarding acute lenticular volume changes in those species manifesting change in lens shape during accommodation. The unusually high colloid osmotic pressure of fish lenses is discussed and related to their unique means of accommodation and their susceptibility to cataract formation. PMID- 7475077 TI - A computer graphic simulation of squamous epithelium. AB - An epithelium maintains its integrity through the organized growth and orderly differentiation of a transient cell population derived from stem cells. This organization is dependent upon both physical mechanisms such as cell adhesion and attraction and the relationship between differentiation and cell division. The interactions between these processes are complex and difficult to conceptualize from a purely mathematical approach. We have therefore set out to develop a graphic model of an epithelium controlled by rules that can be modified. We have chosen to model epidermis, the most superficial part of skin, with cells differentiating from a stem cell population and being lost from the surface of the model. The model is novel not only in the rules that govern cell behaviour, but also because it does not require a predefined lattice to assign the position of cells. Each cell assumes a position depending upon the balance of adhesive and repulsive forces that it experiences. Chemical factors which affect the differentiation of individual cell types are assumed to be produced both by cells within the model and externally from the underlying connective tissue. These "chemical factors" diffuse through the model with a concentration that declines as an inverse square with distance from the source. The rules allow the model to grow from a single stem cell to reach a steady state. At steady state the pattern and clonal structure is strikingly similar to that seen in a range of normal epithelia. Furthermore, if part of the model is removed it is capable of regenerating itself without additional rules. The model allows the visualization of the effects of introducing new rules and modifying the interaction between chosen rules. This study demonstrates that a set of simple rules can be used to make a dynamic flexible model resembling skin. PMID- 7475078 TI - Blood urea clearance with microencapsulated urease. AB - The response of a kidney patient to treatment using a microencapsulated urease artificial kidney (MUAK) system was modelled. The model was used to simulate the patient's response and reactor performance for an initial blood urea concentration of 10 mM and a MUAK void volume of 0.5. The performance of the reactor was strongly dependent on the enzyme activity. An optimal activity of 10 mM sec-1 was achieved in the analysis. After operation for 4 h at a flow rate of 200 ml min-1, the reduction in blood urea concentration for reactor dimensions of 2 x 10 cm, 2 x 20 cm, 4 x 10 cm and 4 x 20 cm were 38%, 52%, 60% and 62%, respectively. The effect of flow rate on the performance of urea removal was studied using the reactor dimensions of 4 x 10 cm (optimal design). The results for flow rates of 100, 200, 300 and 400 ml min-1 predicted blood urea reductions of 38, 60, 70 and 76%, respectively. Although, the conversion of urea in the reactor decreased from 100% to 96% for the respective flows of 100 to 400 ml min 1, the high turnover of reactor volume at a higher flow rate was responsible for the improved reduction of the patient's blood urea level. The model has the ability to predict the performance of the MUAK and the patient's blood urea level simultaneously, at various operating conditions. PMID- 7475079 TI - A note on population dynamics. AB - Population dynamics is analyzed by means of the power series expansion of a decay ratio F(P) with respect to the continuation probability, P, of survival. The truncation of the higher terms of the series yields the Verhulst equation as a first approximation. The approximation for higher age yields a simple exponential decay law of population, while the younger-age approximation recovers Gompertz's empirical law of human mortality. It is shown that there exists a finite survival probability in the limit of t = infinity. The validity of the present result is examined with real population dynamics of centenarians. In order to construct a commensurable definition of aging, an aging phenomenon in decay process is considered on the assumption of an ideal society, from which a simple relationship is derived between the extent of advancement of aging and the continuation probability. PMID- 7475080 TI - Immune responses against multiple epitopes. AB - The current understanding of antigenic escape dynamics is based on models with single epitopes. The usual idea is that a mutation which enables a pathogen (virus, bacteria, etc) to escape from a given immune response confers a selective advantage. The "escape mutant" may then increase in abundance until it induces a new specific response against itself. In this paper a new picture is developed, based on mathematical models of immune responses against several epitopes; the simplest such models can have very complicated dynamics, with some surprising features. The emergence of an escape mutant can shift the immunodominant response to another epitope. Even in the absence of mutations, antigenic oscillation is found, with distinct peaks of different virus variants and fluctuations in the size and specificity of the immune responses. The model also provides a general theory for immunodominance in the presence of antigenic variation. Immunodominance is determined by the immunogenicity and by the antigenic diversity of the competing epitopes. Antigenic oscillations and fluctuations in the cytotoxic T-lymphocyte response have been observed in infections with the human immunodeficiency virus (HIV). Shifting the immune responses to weaker epitopes can represent a mechanism for disease progression based on evolutionary dynamics and antigenic diversity of the virus. PMID- 7475081 TI - George Price's contributions to evolutionary genetics. AB - George Price studied evolutionary genetics for approximately seven years between 1967 and 1974. During that brief period Price made three lasting contributions to evolutionary theory; these were: (i) the Price Equation, a profound insight into the nature of selection and the basis for the modern theories of kin and group selection; (ii) the theory of games and animal behavior, based on the concept of the evolutionarily stable strategy; and (iii) the modern interpretation of Fisher's fundamental theorem of natural selection, Fisher's theorem being perhaps the most cited and least understood idea in the history of evolutionary genetics. This paper summarizes Price's contributions and briefly outlines why, toward the end of his painful intellectual journey, he chose to focus his deep humanistic feelings and sharp, analytical mind on abstract problems in evolutionary theory. PMID- 7475082 TI - The nature of selection. (Written circa 1971, published posthumously) AB - A model that unifies all types of selection (chemical, sociological, genetical, and every other kind of selection) may open the way to develop a general "Mathematical Theory of Selection" analogous to communication theory. [Note added by S. A. Frank: This previously unpublished manuscript was found among Dr. Price's papers when he died in 1975. In this paper Dr. Price did not provide a complete, general theory of selection. Rather, he argued why such a theory is needed and what some of its properties might be. The accompanying article provides commentary on this paper and describes Dr. Price's significant contributions to evolutionary genetics (S. A. Frank, 1995, J. theor. Biol. 175, 373-388).] PMID- 7475083 TI - Quantal transmission at purinergic synapses: stochastic interaction between ATP and its receptors. AB - Monte Carlo methods are used to analyze the stochastic interaction between ATP, released in a packet at a bouton, and the underlying patch of purinoceptors. The time-course of the average quantal current recorded with an intracellular electrode in the peripheral and central nervous systems is reconstructed, given the geometry of the synapse and the known kinetics of ATP action. This leads to certain restrictions on the possible numbers of ATP molecules in a quantum and the density of purinoceptors at the synapses. The addition of an ectoenzyme into the synaptic cleft with the known kinetics of ATPase gives rise to a quantal current of appropriate time-course if the number of ATP molecules in a quantum is increased over that in the absence of the ATPase. The stochastic variability in the quantal current is less than 0.1 for a given size quantum. PMID- 7475084 TI - How much are homologous peptides homologous? AB - A statistical analysis designed to deal with the problem of identifying homologous pairs of "short sequences of amino-acids" (= peptides) belonging to different proteins is presented. The conceptual novelty of the searching strategy proposed here lies in the fact that both the degree of homology of the two peptides of the pair (measured by a suitably defined affinity score) and the level of statistical significance of its occurrence are taken into account on equal footing. They are combined in a sort of "biological indicator", characterising each pair. Pairs for which the value of the biological indicator is larger than an appropriate threshold are taken as statistically significant and (putatively) biologically relevant. The method is employed in various test cases and proves to be reliable and efficient. In particular we have studied the cases in which the known existence of an auto-immune response has lead to the identification of homologous peptide pairs between human and viral or bacterial proteins. The detection efficiency of the algorithm in these cases turns out to be especially good when the most naive affinity table, the Identity matrix, is employed to measure the similarity of amino acidic pairs. In contrast, when the 250-PAM mutation matrix is used, the detection efficiency goes to zero. PMID- 7475085 TI - Search of hidden periodicities in DNA sequences. AB - The hidden periodicities in various genomes can be identified via a Fourier transform of genomic DNA sequences. The typical search strategies are described and the corresponding statistical criteria are presented. The methods are applied to the structural analysis of genome for bacteriophage PHIX174. The results indicate the important role of the coherent structural modifications along the several main periods in the formation of the genome. Some possible mechanisms for the features observed are discussed. PMID- 7475086 TI - The effect of mating structure and progeny distribution on heterozygosity versus the number of alleles as measures of variation. AB - Heterozygosity and the number of alleles are both measures of the genetic variation of a population. They are qualitatively similar if the distribution of the number of progeny is Poisson, but not necessarily for other distributions. In particular, selfing decreases heterozygosity and also decreases the number of alleles when the progeny distribution is Poisson, but decreases heterozygosity and increases the number of alleles when there are two progeny per individual. This is because heterozygosity is closely related to the breeding structure (inbreeding) of the population, whereas the number of alleles is more influenced by the variation of the sampling process. Branching processes are employed to model the dynamics of mutant alleles, with drift and subsequent mutation contributing to extinction. In populations of four individuals, double first cousin mating has greater heterozygosity, but fewer alleles, than half-sib mating. PMID- 7475087 TI - Simple physical constraints in hemolysis. AB - The percentage of normal, red blood cells that are hemolyzed when placed in hypotonic solutions depends on a variety of factors, two important ones being the initial sphericities of the cells and the tonicities to which they are subjected. Other, less well-understood factors that are important in hemolysis are the initial cell volumes, how much free water they contain and the elasticity of the cell membranes. The purpose of this work is to identify the constraints a red cell must satisfy in order to be hemolyzed. Human erythrocyte data is used in a physical model that compares the balance of hydrostatic stresses in sphered cells that are on the verge of hemolysis. For hemolysis to occur we find there is a critical sphericity index that must be exceeded. It depends on tonicity, the initial, fractional water volume in the cells and the maximum fractional area dilation the cell membranes can withstand. Membrane tensile strength and the non ideal osmotic behavior of hemoglobin are of relatively minor importance. But when they are taken into account, the hemolysis constraint, in the form of a remarkably simple inequality, compares favorably with clinical tests of erythrocyte osmotic fragility. PMID- 7475088 TI - Drosophila melanogaster syncytial nuclear divisions are patterned: time-lapse images, hypothesis and computational evidence. AB - Time-lapse microscopy of biological systems has provided new and exciting information about the dynamics of cellular and developmental events. However, these events are often complex and difficult to analyze. This paper describes a study in which computation was indispensable for formulating and evaluating a cellular/developmental hypothesis directly from observations of time-lapse fluorescence images. Previous analyses of time-lapse microscopy sequences of Drosophila melanogaster embryonic syncytial nuclear cycles 10-13, when the nuclei form an evenly spaced monolayer at the surface of the embryo, have failed to identify any pattern in these divisions. However, computational analysis of the data has provided evidence that the direction of syncytial nuclear mitosis is not random, but is clearly influenced by the relative positions of neighboring nuclei. An approximate law governing mitotic direction that is based on a scheme that compromises among "votes" made by neighboring nuclei is introduced. PMID- 7475089 TI - Analytical solutions of the dinucleotide probability after and before random mutations. AB - The mutation process is a classical evolutionary genetic process mainly based on the (random) substitutions of one base (A = Adenine, C = Cytosine, G = Guanine, T = Thymine) for another. Two analytical solutions derived here allow us to analyse in genes the occurrence probabilities of motifs (e.g. dinucleotides) after substitutions (in the evolutionary sense: from the past to the present) and, unexpectedly, also before substitutions (after back substitutions, in the inverse evolutionary sense: from the present to the past). We generalize on the alphabet [A, C, G, T] of the analytical solutions and of the properties derived on the alphabet [R, Y] (R = purine = A or G, Y = pyrimidine = C or T). Application of the theory is based on the analytical solution giving the probabilities of the 16 dinucleotides AA, ..., TT in the protein (coding) genes of (nuclear) eukaryotes, viruses and prokaryotes and in (eukaryotic) introns after back substitutions (called primitive genes). After back substitutions, four of 16 dinucleotides--CG, TA, GT and AC--occur with low probabilities in each of these four primitive gene populations, except for CG in the primitive prokaryotic protein genes. In the primitive eukaryotic protein genes, the dinucleotide AT has also a significant low probability. We present the properties of the two analytical solutions, and the functions which may have these five dinucleotides in primitive genes are described in terms of biological signals. PMID- 7475090 TI - Characteristics of atrial re-entry and meander computed from a model of a rabbit single atrial cell. AB - A two-dimensional excitable medium model of atrial tissue has been constructed by incorporating the Earm-Hilgemann-Noble excitation equations for membrane voltage dependent ionic currents and pump exchanger currents, time-dependent changes in intracellular and extracellular ionic concentrations, and storage and release of Ca2+, for an atrial cell in an homogeneous partial differential equation. A diffusion coefficient of 1.25 cm2 sec-1 gives the conduction velocity of a solitary planar wave as 0.6 m sec-1. A spiral wave with a period of 78 msec develops from a broken wavefront, and initially rotates around a circular core of 2 mm diameter. After 1.5 sec of rotation, a biperiodic meander, with 6 epicycles/cycle, develops. This simple meander pattern is reflected in a period-6 modulation of the intervals between successive wavefronts. PMID- 7475091 TI - Population dynamics of feline immunodeficiency virus within cat populations. AB - A deterministic model was constructed for studying the circulation of Feline Immunodeficiency Virus (FIV), a feline retrovirus homologous to Human Immunodeficiency Virus (HIV), within populations of domestic cats. The model has been tested with data generated by a long-term study of several natural cat populations. Simulations and a study of stability show that once introduced, the retrovirus is maintained within the population, with a stable equilibrium stage reached by both numbers of susceptible and infected individuals. An estimation of parameters indicates that the transmission rate is low and depends on the structure of the population. In addition, FIV has a low impact on the population in that the total number of cats at equilibrium when this virus is present is almost always equal to the habitat carrying capacity in the absence of the virus. Those results, in agreement with other observations, suggest that FIV originally arose in the distant past. PMID- 7475092 TI - Towards a general function describing T cell proliferation. AB - A new function is proposed for describing the rate of T cell proliferation in response to peptides on antigen-presenting cells. The model improves an earlier model of ours by allowing for a true maximum proliferation rate of the T cells. This is achieved by a simple change of variables that markedly relaxes the conditions for a conventional quasi-steady-state assumption. The new model has the same "ecological" properties as the previous one. Thus the natural competition in the model allows for regulation of T cell population size in the presence of continuous stimulation by antigen. An important feature is the competitive exclusion of T cell clones recognizing the same peptide with different affinities allowing for "affinity selection". Models for the population dynamics of experienced, naive and activated T cells are also developed. These T cell subpopulations compete with one another for antigen. In models with lymphokine production a "proliferation threshold" is obtained that allows for tolerance. PMID- 7475093 TI - A model of non-exclusive binding of agonist and antagonist on G-protein coupled receptors. AB - A modified concept of functioning of G-protein coupled receptors is presented, on the assumption that agonistic and antagonistic effects of drugs are related to their interaction with two separate receptor sites that exist simultaneously on a single receptor molecule and possess different ligand-specificity patterns. This proposal distinguishes between agonists and antagonists as binding at one of these sites triggers the receptor response, whereas the other site elicits another response leading to the receptor blockade. As these sites are simultaneously present on a single receptor molecule the formation of a ternary complex between agonist, antagonist and receptor is possible. Practical consequences of this concept are analysed with reference to experimental data on muscarinic acetylcholine receptors. PMID- 7475094 TI - Contamination of the genome by very slightly deleterious mutations: why have we not died 100 times over? AB - It is well known that when s, the selection coefficient against a deleterious mutation, is below approximately 1/4Ne, where Ne is the effective population size, the expected frequency of this mutation is approximately 0.5, if forward and backward mutation rates are similar. Thus, if the genome size, G, in nucleotides substantially exceeds the Ne of the whole species, there is a dangerous range of selection coefficients, 1/G < s < 1/4Ne. Mutations with s within this range are neutral enough to accumulate almost freely, but are still deleterious enough to make an impact at the level of the whole genome. In many vertebrates Ne approximately 10(4), while G approximately 10(9), so that the dangerous range includes more than four orders of magnitude. If substitutions at 10% of all nucleotide sites have selection coefficients within this range with the mean 10(-6), an average individual carries approximately 100 lethal equivalents. Some data suggest that a substantial fraction of nucleotides typical to a species may, indeed, be suboptimal. When selection acts on different mutations independently, this implies too high a mutation load. This paradox cannot be resolved by invoking beneficial mutations or environmental fluctuations. Several possible resolutions are considered, including soft selection and synergistic epistasis among very slightly deleterious mutations. PMID- 7475095 TI - Alpha-fetoprotein signal sequences: a proposed mechanism for subcellular localization and organelle targeting. AB - Alpha-fetoprotein (AFP) is an oncofetal protein, classified in an "albuminoid" superfamily (with albumin and Vitamin-D binding (Gc) protein) comprising molecules with three characteristic globular domains. The cellular uptake and internalization of AFP and its subcellular compartmentalization has previously been reported in a multitude of cell types. Studies have also emerged which have detected and characterized binding proteins complexed to AFP in various cell membranes and intracellular compartments. However, the literature is devoid of any attempts to relate these binding proteins to possible intracellular trafficking interactions of AFP. Recombinant DNA mutation/deletion technology has provided a means to pinpoint the amino acid sequence location of organelle localization signals on various transcription factors and/or receptors. Several subdomain regions on AFP have been reported to mimic heptad dimerization regions of the steroid/thyroid nuclear receptors. In light of these transcription factor like docking motifs reported for AFP, the present report purposes various subdomain regions which might constitute basic amino acid sequences resembling recognition signals for binding (dimerizing) proteins. AFP appears to possess multiple prototypic amino acid sequence cassettes on each domain which consist of (i) classical short, compact sequences found on both steroid and thyroid receptors; (ii) proto-signals resembling a steroid receptor type; and (iii) degenerative signal sequence similar to the thyroid/retinoic acid receptor type. The concepts identifying binding or escort proteins for AFP together with organelle signal sequences on AFP have been reconciled in a composite hypothesis to formulate a mechanism which could account for some of the growth-regulating properties described for AFP during the last decade. PMID- 7475096 TI - Michaelis-Menten mechanism reconsidered: implications of fractal kinetics. AB - The Michaelis-Menten formalism assumes that the elementary steps of an enzymatic mechanism follow traditional mass-action kinetics. Recent evidence has shown that elementary bimolecular reactions under dimensionally-restricted conditions, such as those that might occur in vivo when reactions are confined to two-dimensional membranes and one-dimensional channels, do not follow traditional mass-action kinetics, but fractal kinetics. A Michaelis-Menten-like reaction operating under conditions of dimensional restriction is shown to exhibit new types of synergism and noninteger kinetic orders. These properties are likely to have an important influence on the behavior of intact biochemical systems, which is largely dependent upon the kinetic orders of the constituent biochemical reactions. PMID- 7475097 TI - A speed limit for evolution. AB - An upper bound on the speed of evolution is derived. The bound concerns the amount of genetic information which is expressed in observable ways in various aspects of the phenotype. The genetic information expressed in some part of the phenotype of a species cannot increase faster than a given rate, determined by the selection pressure on that part. This rate is typically a small fraction of a bit per generation. Total expressed genetic information cannot increase faster than a species-specific rate--typically a few bits per generation. These bounds apply to all aspects of the phenotype, but are particularly relevant to cognition. As brains are highly complex, we expect large amounts of expressed genetic information in the brain--of the order of 100 kilobytes--yet evolutionary changes in brain genetic information are only a fraction of a bit per generation. This has important consequences for cognitive evolution. The limit implies that the human brain differs from the chimpanzee brain by at most 5 kilobytes of genetic design information. This is not enough to define a Language Acquisition Device, unless it depends heavily on pre-existing primate symbolic cognition. Subject to the evolutionary speed limit, in changing environments a simple, modular brain architecture is fitter than more complex ones. This encourages us to look for simplicity in brain design, rather than expecting the brain to be a patchwork of ad hoc adaptations. The limit implies that pure species selection is not an important mechanism of evolutionary change. PMID- 7475098 TI - The frequency response of ion channel currents in a combined diffusion and barrier type model. AB - The dynamics of ion transport through channels has been calculated using a small signal analysis method. This is applied to the equations describing the rates of ion transport through channels represented by a pair of entrance barriers in series with an interior diffusion regime. The responses to combinations of applied DC and AC electric fields, assumed to be constant in space, have been calculated and are represented as complex admittances. On examining the admittance properties, the channel is found to behave electrically as a double reactance network, which can give rise to resonance effects. A further interesting property is the limiting slope of the current amplitude at high frequencies, which is found to exhibit an f-2/3 dependence for small entrance barriers and an f-1 dependence for large entrance barriers. This points to the possibility of using noise measurements for drawing conclusions about ion transport properties through channels. PMID- 7475099 TI - The use of regular isotopy in the topological description of site-specific DNA recombination processes. PMID- 7475100 TI - The ontogeny of Gaia: the role of microorganisms in planetary information network. AB - The Gaia theory states that the whole surface layer of the globe, including organisms, atmosphere, hydrosphere and lithosphere, can be viewed as a self evolving homeostatic system. Bacteria play a principal role in this formative process. This line of thinking should answer several questions: Is the biosphere simply a set of essentially selfish individuals, each testing its fitness? Is memory in the biosphere confined to genes? Is the composition of ecosystems and their behavior only kind of dynamic equilibrium? Or is there a huge amount of epigenetic information that enables organisms to interpret properly their situation and behave accordingly? It is suggested here that it is the information flow, rather than pure thermodynamics and natural selection, that enables living beings not only to "read" their environment, interpret their situation and behave accordingly to this understanding, but also to establish geophysiology, to become part of Gaia. This flow comprises genes as well as signals, diffusible as well as permanent. From an analogy with the situation in the multicellular organism, three types of information mediators inside the body of Gaia are suggested: ultrastructure, diffusible signals and gene flux. With a shared gene pool, intricate ultrastructure and quickly diffusible signals, it is legitimate to consider Gaia a superorganism with an ontogeny. PMID- 7475101 TI - Microscopic self-organization in living cells: a study of time matching. AB - Biochemical subsystems of a living cell may operate with only a few thousand enzyme molecules and their response can be triggered by the entrance of individual molecules of a certain species. This mode of operation is not described by classical chemical kinetics, which deals with large numbers of reacting molecules. Theoretical estimates for the characteristic times of enzymic reactions in small cells and cellular compartments show that any two macromolecules within a micrometer-size volume meet each other each second and that the transit time, required for a mediator molecule to meet a target enzyme, is comparable to the duration of a catalytic round for a single enzyme molecule. When these conditions are satisfied, an enzymic subsystem represents a coherent molecular network with persistent strong temporal correlations between the catalytic events of individual enzyme molecules. PMID- 7475102 TI - Fluctuations in living cells and intracellular traffic. AB - Theoretical estimates reveal the presence of strong thermal hydrodynamic fluctuations in the cytoplasm of living cells. It is suggested here that these intensive fluctuations may be utilized by biological vesicles inside the cells to produce their directed drift to the targets. A possible mechanism of partial rectification of thermal fluctuations is based on the effect of active navigation: i.e. on the controlled response of vesicles to velocity fluctuations. PMID- 7475103 TI - Homeostasis perturbation (physiological stress) and the mechanism of biological evolution--the possible role of active oxygen species. PMID- 7475104 TI - The evolutionarily stable strategy under individual condition and tactic frequency. AB - A proof is presented to show that, when fitnesses from alternative tactics within a population depend on both their frequency and the phenotypic condition of individuals, there will be a unique ESS switchpoint s* that determines both the condition at which an individual will switch between tactics and the resulting frequency of the tactics in the population. For an individual at the ESS s*, the fitnesses of the alternative tactics will be equal. When fitness is averaged over the population, however, the average fitnesses of the alternative tactics will not be equal. PMID- 7475105 TI - Compact and scattered gap junctions in diffusion mediated cell--cell communication. AB - Gap junctions have been thought of as prime sites for the passive communication of small molecules between two adjacent cells. The sizes of a gap junction plaque may range from a few junctional particles to several thousands particles on the interface of two cells, with little being known about the relative contribution of different patterns of junctional plaques on diffusion mediated cell--cell communication. In this paper, by constructing a simple cubic lattice model of a two-cell system, the effects on communication efficiency of two extreme distribution patterns of individual junctional particles rae investigated. This model indicates that scattered gap junctions are an order of magnitude higher in communication efficiency than compact junctional plaques of equal number of total junctional particles. Thus, it is possible that modulation of cell-cell communication may rely on the interconversion of these two states with compact plaques existing as an inactive communication form. Any correlation of gap junctional membrane with cell-cell communication should therefore also take into account its configuration. It is possible that an increase in number and area of tightly packed gap junctional plaques may actually decrease, instead of increase, cell-cell communication. PMID- 7475106 TI - A generalized epistasis analysis as a method of reconstruction of developmental process and its application to Drosophila dorso-ventral axis formation. AB - Epistasis analysis has been used in molecular genetics to infer cause and effect relationships among loci that decide a developmental fate. In this paper, in order to investigate the logical background of the rule that relates epistasis and causal relation, a mathematical epistasis analysis is formulated. An "orderliness index" is introduced by which the cause-and-effect relation can systematically be extracted from genotype-phenotype data. This method is applied to Drosophila melanogaster dorso-ventral axis formation and it is shown that the method of reconstructing the framework of the causal relation between a locus and its sign (activation or inhibition) as an "orderliness diagram" from eight groups of the genes. From the diagram it can be seen that the dorsal gene works as a morphogen. The difference between orderliness diagrams of the dorsal and ventral sides of the embryo shows that the cells on the ventral side receive the signal and the fate of the cells on the dorsal side is default in the wild-type embryo. PMID- 7475107 TI - Physiological time and the measurement of glomerular filtration rate. AB - Diverse physiological timescales in different animals can have an important influence on the measurement of biological variables. In vertebrates, the absolute and relative rates of glomerular filtration span several orders of magnitude. These differences have important implications for the logistics of measurement protocols in different species. A simple model is used to examine the importance of physiological time to the measurement of glomerular filtration rate. PMID- 7475108 TI - A model of the virus-type strategy in the early stage of encoded molecular evolution. AB - Recent advances in evolutionary molecular engineering have revealed that the essential nature of a "virus" in the evolutionary aspect is its bonding strategy for assignment of the phenotype to its genotype. Based on the definition of "virus"-type and "cell"-type of the assignment strategy, we propose a virus early/cell-late model of the history of life. The first encoded protein is assumed to be a cofactor of replication ribozyme in the RNA world and to be bound to its genetic RNA. As such a virus-type strategy could introduce the Darwinian selection process into the hypercycle with translation, a hypercycle with virus like members could make the replicase protein and the translation system gradually evolve together out of the RNA world without a proto-cell. Moreover, they could evolve much faster by this virus-type strategy than by a primitive cellular organism. PMID- 7475109 TI - Cellular segregation and engulfment simulations using the cell programming language. AB - In developmental biology, modeling and simulation play an important role in understanding cellular interactions. In this paper a simple language, the Cell Programming Language (CPL), is suggested for writing programs that describe this behavior. Using these programs, it is possible to simulate and visualize intercellular behavior. CPL is used to model cellular segregation based upon the differential adhesion hypothesis. Results indicate that a high degree of segregation can be produced in a mixture of cells by allowing random motion. The engulfment of a tissue by a less adhesive tissue is also observed when the two tissues are placed in contact. Both these simulations utilize only local interactions and random motion of cells. Earlier simulations used long-range interactions to observe similar effects. The present simulations prove that random motion of cells can produce long-range effects. PMID- 7475110 TI - Evolutionary cycling in predator-prey interactions: population dynamics and the red queen. AB - This paper describes the coevolution of phenotypes in a community comprising a population of predators and of prey. It is shown that evolutionary cycling is a likely outcome of the process. The dynamical systems on which this description is based are constructed from microscopic stochastic birth and death events, together with a process of random mutation. Births and deaths are caused in part by phenotype-dependent interactions between predator and prey individuals and therefore generate natural selection. Three outcomes of evolution are demonstrated. A community may evolve to a state at which the predator becomes extinct, or to one at which the species coexist with constant phenotypic values, or the species may coexist with cyclic changes in phenotypic values. The last outcome corresponds to a Red Queen dynamic, in which the selection pressures arising from the predator-prey interaction cause the species to evolve without ever reaching an equilibrium phenotypic state. The Red Queen dynamic requires an intermediate harvesting efficiency of the prey by the predator and sufficiently high evolutionary rate constant of the prey, and is robust when the model is made stochastic and phenotypically polymorphic. A cyclic outcome lies outside the contemporary focus on evolutionary equilibria, and argues for an extension to a dynamical framework for describing the asymptotic states of evolution. PMID- 7475111 TI - Fixation of epigenetic states in a population. AB - Multiple steady-state levels of autoregulatory transcription factors constitute alternative epigenetic states, and could help specify the heritable phenotype of an organism. In this work, the inheritance of autoregulatory transcription factors through sexual reproduction is modeled to determine the conditions under which different alternative epigenetic states become fixed in a population. The influence of transient environmental perturbations on the fixation of states is discussed in the context of speciation. PMID- 7475112 TI - Status signaling games in animal contests. AB - This paper provides evolutionary game-theoretic models for the status signaling problem in animal contests. Two stage status signaling game models are introduced and evolutionarily stable strategies and their modifications are characterized for the games. It is shown that in the stable long-run equilibria, animals must make use of signals systematically to settle the contest peacefully. PMID- 7475113 TI - Simulation of chromosomal homology searching in meiotic pairing. AB - A cellular automaton model has been developed to simulate some aspects of chromosome behaviour during meiotic prophase when homologous chromosomes search for each other so that they can pair. Simulations allow one to compare the relative efficiencies of random searching by chromosome shuffling along the inner nuclear membrane or within the lumen of the nucleus, the effects of biased movements and of chromosome clustering (bouquet or synizetic knot), different numbers of homology recognition sites per chromosome and different chromosome numbers on the efficiency of the pairing process. Parameters of the simulated models can be easily adjusted to fit experimentally obtained figures. PMID- 7475114 TI - Intracellular transport mechanisms: a critique of diffusion theory. AB - It is argued that Brownian motion makes a less significant contribution to the movements of molecules and particles inside cells than is commonly believed, and that the numbers of similar molecules and particles within any near-homogeneous subcompartment of the cell internum are insufficient to justify the statistical assumptions implicit in the derivation of the diffusion equation. For these reasons, it is contended that, contrary to accepted opinion, diffusion theory cannot provide an explanation for intracellular transport at the molecular level. Although attempts have been made to adapt diffusion theory to complex media, the conclusion is that none satisfactorily overcomes the problem of applying the theory to cell biology. However, the heuristic influence of the theory on cellular biophysics and physiology is noted, and possible alternative frameworks for interpreting the valuable experimental data obtained from such studies are outlined. PMID- 7475115 TI - The developmental asynchrony hypothesis for sex ratio manipulation. AB - This paper presents the hypothesis that developmental asynchrony between embryos of both sexes, on the one hand, and offspring development and responsiveness of maternal reproductive organs, on the other, may result in significant sex ratio biases at birth in mammals, and in birds. It is argued that the developmental asynchrony hypothesis may account for a significant proportion of findings on sex ratio skews, including the inconsistency of evidence and the lack of successful selection attempts. This mechanism could be used by the maternal organism adaptively to alter the sex ratio by control of the relative time of insemination within the oestrous cycle or by alteration of reproductive hormone levels around the time of ovulation and early embryonic development. However, the developmental asynchrony hypothesis may also imply that many of the sex ratio biases can be viewed as by-products of constraining selection on control of fertility and fecundity and on sexual dimorphic growth rates. PMID- 7475116 TI - The dynamics of Helicobacter pylori infection of the human stomach. AB - Helicobacter pylori is a bacterial pathogen of the gastrointestinal tract of humans causing chronic superficial gastritis which persists for decades. The mechanism by which H. pylori is able to persist, despite environmental constraints, remains unknown. Therefore, a model is proposed describing the interactions of H. pylori with its host, involving an autoregulatory network in which inflammation leads to nutrient release. A determinist mathematical model examining the interactions necessary to maintain chronic infection indicates that this proposed autoregulatory network can produce steady-state solutions, and the model is robust in encompassing biological variations. PMID- 7475117 TI - A mathematical framework for describing and analysing gene regulatory networks. AB - This paper presents a mathematical framework for describing and analysing gene regulatory networks by autonomous differential equations. It represents an improvement on existing frameworks in that it may handle a wider range of gene regulatory mechanisms. Gene regulatory networks are frequently threshold dominated, i.e. genes are activated only when the concentration of certain gene products lie between definite thresholds. Here, the concept of regulatory domain is introduced to describe these regions in the phase space. To each regulatory domain is associated an indicator function whose value is 1 inside and 0 outside the domain. The indicator functions thus reflect the logical structure of the network. The sharp borders between the regulatory domains may be smoothed by replacing the logical step functions by continuous sigmoids or so-called logoid functions. A logoid function coincides with the step function outside a narrow interval around the threshold, and rises continuously from 0 to 1 inside it. Using logoids, the task of finding steady states is considerably simplified. A list of regions in phase space comprising all steady states lying close to a threshold is obtained by examining a certain type of matrix called the Logoid Jacobian. In addition, this matrix leads to the conditions necessary for stability of the steady states. External signals may be conveniently incorporated in the form of Boolean variables. Thus the framework is well suited for studying gene regulatory networks both in single cells and multicellular systems. PMID- 7475118 TI - Anti-ulcerogenic evaluation of the methanolic extracts of some indigenous medicinal plants of Pakistan in aspirin-ulcerated rats. AB - Anti-ulcerogenic activity of the methanolic extracts of 4 medicinal plants were studied in aspirin-induced gastric ulcers in rats. Their effects on the volume of gastric juice secreted, acid output, peptic activity, mucin activity and curative ratio were recorded. Bauhinia racemosa (flower buds) decreased the ulcer index significantly, and Moringa pterygosperma (flower buds) showed some decrease in the ulcer index. Trianthema pentandra (whole plant) did not show any decrease in the acid or pepsin content or any increase in mucin; however, it showed a highly significant decrease in the ulcer index. Cordia latifolia (ripened fruit) did not however decrease the ulcer index. PMID- 7475119 TI - Anti-giardial activity of gastrointestinal remedies of the Luo of east Africa. AB - Activity in an in vitro assay with Giardia lamblia provided a test of the validity of a quantitative methodology used in an ethnobotanical survey of the Luo people of the Lake Victoria basin of Kenya and Tanzania. Forty-five taxa of remedies for gastrointestinal problems were reported by four or more independent informants and a log-linear model was used to calculate a statistical measure of informant consensus. Methanolic extracts of 21 of 36 taxa assayed were lethal or inhibited growth of Giardia trophozoites at 1000 ppm; 7 species were lethal at 500 ppm. Non-cathartic species are more likely to be active than cathartics. Lethal species of non-cathartics are reported by informants more frequently than non-lethal species although the lack of statistical significance did not provide satisfactory support for the validity of the quantitative methodology as a predictor of efficacious remedies. PMID- 7475120 TI - Xanthine oxidase inhibitory activity of some Panamanian plants from Celastraceae and Lamiaceae. AB - Thirty four crude extracts of Panamanian plants, from nine species of Celastraceae and Lamiaceae, were assayed for xanthine oxidase (XO) inhibitory activity. The enzymatic activity was estimated by measuring the increase in absorbance at 290 nm due to uric acid formation. Eighty five percent of the crude extracts were found to possess XO inhibitory activity at 50 micrograms/ml and all the extracts of the species from Lamiaceae were active even at 1 micrograms/ml. The ethanol extracts of Hyptis obtusiflora Presl ex Benth. (Lamiaceae) and H. lantanaefolia Poit. (Lamiaceae) exhibited the highest activity with an inhibition of approximately 40% at 1 micrograms/ml. PMID- 7475121 TI - Screening of hundred Rwandese medicinal plants for antimicrobial and antiviral properties. AB - A series of 100 Rwandese medicinal plants (267 plant extracts), used by traditional healers to treat infections, were screened for antibacterial, antifungal and antiviral properties. The results of the testing showed that 45% were active against Staphylococcus aureus, 2% against Escherichia coli, 16% against Pseudomonas aeruginosa, 7% against Candida albicans, 80% against Microsporum canis and 60% against Trichophyton mentagrophytes. Not less than 27% of the plant species exhibited prominent antiviral properties against one or more test viruses, more specifically 12% against poliomyelitis, 16% against coxsackie, 3% against Semliki forest, 2% against measles and 8% against herpes simplex virus. PMID- 7475123 TI - Effect of different formulations of propolis on mice infected with Trypanosoma cruzi. AB - Propolis, a bee product, can be regarded as one of the potential natural sources in folk medicine, displaying strong antimicrobial activity. Previous work showed that propolis extracts exhibited in vitro activity against Trypanosoma cruzi (Higashi and de Castro, 1994). Different formulations of propolis were administered to experimentally Trypanosoma cruzi-infected mice and the parasitemia kinetics and survival rate were monitored. The oral administration of ethanolic extracts up to 1.2 g propolis/kg per day or propolis offered ad libitum in the drinking water (up to 4 g/kg per day) or added to the food (up to 5 g/kg per day) did not interfere with both parameters. The differences between in vitro and in vivo trypanocidal activity of propolis and future perspectives are discussed. PMID- 7475122 TI - Pharmacological effects of dai-saiko-to on lipid biosynthesis in cultured human hepatocyte HepG2 cells. AB - The pharmacological effects of Dai-saiko-to, a Japanese and Chinese traditional medicinal mixture (Kampohozai), on lipid biosynthesis were investigated in cultured human hepatocyte HepG2 cells. The addition of Dai-saiko-to (0.5 mg/ml), which had no significant effect on cell proliferation, caused a marked decrease in the intracellular triglyceride content with no significant changes in the other lipid fraction. At the same time, the incorporation of 14C-acetate or 3H glycerol into the triglyceride or diglyceride fractions also decreased significantly. These results suggest that Dai-saiko-to decreases hepatic triglyceride biosynthesis, which might contribute to a reduction in plasma VLDL levels. PMID- 7475124 TI - Inhibitory effects of Cistus populifolius on contractile responses in the isolated rat duodenum. AB - The medicinal plant Cistus populifolius L., shows an important dose-dependent spasmolytic activity. The ability of the Cistus extract to inhibit both acetylcholine (ACh) (3.4 x 10(-8)-6.8 x 10(-5) M) and CaCl2 (2 x 10(-4)-1.28 x 10(-2) M)-induced contractions and the relaxing effect on K+ (75 mM)-induced contractions may indicate a non-specific receptor antagonist. However, this action may be related to the influx of extracellular Ca2+. These antispasmodic effects are partly consistent with the use of C. populifolius in folk medicine for certain gastrointestinal disorders. PMID- 7475125 TI - Vasoactive effects of aqueous extracts from five Mexican medicinal plants on isolated rat aorta. AB - The present investigation describes the effects of aqueous extracts from Chiranthodendron pentadactylon flowers, Galphimia glauca leaves and flowers, Ipomoea stans roots, Juglans regia leaves and Taxodium mucronatum aerial parts on isolated rat thoracic aorta precontracted by noradrenaline (NA). In all cases, the aqueous extracts (0.5-12 mg/ml) significantly inhibited, in a concentration dependent manner, the maximal contractile response induced by NA in rat aorta. The most active extract was that of G. glauca flowers. These findings indicate that the active principle(s) present in the crude extracts can exert a vasorelaxant effect. PMID- 7475126 TI - Antiinflammatory activity of a Ghanaian antiarthritic herbal preparation: III. AB - alpha-Amyrin palmitate, present in a Ghanaian antiarthritic herbal preparation of Alstonia boonei, Elaies guineensis and Rauvolfia vomitoria, was synthesised and tested on complete Freund's adjuvant-induced arthritic rats. Administered orally at 56 mg/kg body weight (BW) daily for 8 days from days 11 to 18 post adjuvant (acute) or at 66 mg/kg BW every 48 h for 5 days from days 32 to 40 (chronic), the drug returned the increases in serum hyaluronate and blood granulocytes towards non-arthritic levels and corrected the moderate anaemia of adjuvant arthritis. Histological examinations of the proximal interphalangeal foot joints showed reduced synovial proliferation and invasion of joints and reduced leucocyte infiltration of bone marrow and periarticular tissue in treated rats. The results suggest that alpha-amyrin palmitate contributes to the previously shown antiarthritic effect of the herbal preparation. PMID- 7475127 TI - Natural premedication for mast cell proliferative disorders. PMID- 7475129 TI - The role of coronary revascularization in recipients of an implantable cardioverter-defibrillator. AB - The impact of adjuvant coronary revascularization was studied in a group of 138 recipients of an implantable cardioverter-defibrillator, all of whom had ischemic heart disease as the cause of their arrhythmias. Patients chosen for revascularization had more severe anatomic, symptomatic, or physiologic evidence of active ischemia. There were no operative deaths among 23 patients who actually underwent coronary artery bypass combined with cardioverter-defibrillator implantation; however, operative mortality by the intention-to-treat principle was 8% (2/25). Total cardiac survival was better for patients who underwent revascularization than for those patients who had "high-risk" characteristics and did not undergo revascularization. Stratified subgroup analysis demonstrated significant survival advantages favoring revascularization in patients with three vessel or left main coronary artery disease, class III or IV angina, and an ejection fraction greater than 25%. Multivariate analysis revealed that low ejection fraction and left main coronary artery disease were independent predictors of decreased survival. PMID- 7475130 TI - Age-related development of human anti-pig xenoantibody. AB - Human cytotoxic natural xenoantibodies are believed to be of the immunoglobulin M class in nature. However, a thorough understanding of the development of these natural antixenodonor xenoantibodies remains incomplete. In this study, serum samples were obtained from newborn, infant, and adult human beings. An enzyme linked immunosorbent assay was used to determine the binding of human natural immunoglobulin M xenoantibodies to pig aortic endothelial cells and pig lymphocytes. A complement-mediated cytotoxicity assay was used to measure the cytotoxicity of newborn, infant, and adult serum to cultured pig aortic endothelial cells and pig lymphocytes. Adult human serum contained both natural immunoglobulin M and immunoglobulin G xenoantibodies to pig endothelial cells and lymphocytes, whereas newborn infant serum contained only immunoglobulin G xenoantibodies. Only adult human serum was cytotoxic to pig endothelial cells and lymphocytes. Human immunoglobulin M xenoantibodies became detectable by age 1 month. By age 2 months these natural anti-pig xenoantibodies reached serum levels equivalent to those in the human adult and resulted in similar cytotoxicity to that of adult human serum. These findings indicate that (1) natural anti-pig immunoglobulin M xenoantibodies are absent from newborn infant human serum, (2) newborn human serum is not cytotoxic to pig endothelial cells and lymphocytes despite the presence of immunoglobulin G xenoantibodies that bind to pig endothelial cells and lymphocytes, and (3) natural anti-pig immunoglobulin M xenoantibodies begin to develop as early as age 1 month and by age 2 months attain a circulating level comparable to that found in the adult. PMID- 7475128 TI - Reactivity of human saphenous veins at arterial perfusion pressures. AB - Vasospasm of human saphenous vein grafts has been reported after aorta-coronary bypass operations. However, it is unknown whether veno-arterial grafts are inherently responsive to vasoconstrictor stimuli after implantation into the arterial circulation or whether their vasomotion is secondary to hemodynamic changes. Thus in this study we used in vitro methods to directly evaluate whether isolated human saphenous vein segments respond to vasoconstrictor agents at arterial pressure levels. External diameter and intraluminal flow were monitored in 12 human saphenous vein segments, which were perfused at 30 ml/min with physiologic salt solution at 90, 70, and 50 mm Hg. Increasing intraluminal pressure higher than 50 mm Hg or exposing the vein to Ca(2+)-free media did not increase vessel external diameter or intraluminal flow, which suggests that human saphenous veins were fully distended at pressures of 50 mm Hg or greater. However, all human saphenous veins were activated by a 1 mumol/L dose of norepinephrine at 50 mm Hg and dilated during subsequent intraluminal infusion of a 1 mumol/L dose of acetylcholine, showing intact vascular smooth muscle and endothelial cell function. In the same vessels, a 1 mumol/L concentration of 5 hydroxytryptamine constricted human saphenous veins by 19%, 22%, and 26% at intraluminal pressures of 90, 70, and 50 mm Hg, respectively, and reduced vessel flow by 6%, 24%, and 42% at the same pressure levels. Similarly, a 1 mumol/L concentration of norepinephrine constricted vessels pressurized at 90, 70, and 50 mm Hg by 9%, 12%, and 17%, respectively, and attenuated vessel flow by as much as 32%. We conclude that human saphenous vein segments are fully distended at perfusion pressures greater than 50 mm Hg, but can dynamically constrict to vasoactive agonists and regulate graft flow at intraluminal pressures as high as 90 mm Hg. Our findings in isolated human saphenous vein segments lend support to clinical observations that human saphenous vein grafts should be regarded as vasoactive conduits after implantation at arterial pressure levels. PMID- 7475131 TI - Single-stage treatment of sternal wound complications in heart transplant recipients in whom pectoralis major myocutaneous advancement flaps were used. AB - Treatment of sternal wound complications is controversial, particularly in immunosuppressed heart transplant recipients. Regardless of the severity of infection, we combine immediate, aggressive debridement with bilateral pectoralis major myocutaneous advancement flaps in a single procedure. Compared with management with pectoralis major turnover flaps or distant pedicled muscle flaps, treatment of these sternal wounds with pectoralis major myocutaneous advancement flaps is simpler and quicker and provides better aesthetic results. Furthermore, because pectoralis major myocutaneous flaps are based on the thoracoacromial arteries, whether or not the internal mammary arteries have previously been harvested for coronary grafts is irrelevant. Twenty consecutive heart transplant recipients with sternal wound complications were treated with this technique. No intraoperative or perioperative deaths occurred. The morbidity rate was 30%, with seroma treated by needle aspiration in four patients (20%) being the most common complication. Only one patient had a postoperative wound infection. All patients had excellent functional and aesthetic results. PMID- 7475132 TI - Experimental tracheal and tracheoesophageal allotransplantation. Paris-Sud University Lung Transplantation Group. AB - We investigated the effects of allograft perfusion with a preservative technique and of combined thyrotracheoesophageal implantation on airway epithelium of long segments of thyrotracheal grafts allotransplanted on their own vascular pedicles into immunosuppressed pigs. Four groups of five animals each underwent heterotopic (into the neck) thyrotracheal (group 1) and thyrotracheoesophageal (group 2) and orthotopic thyrotracheal (group 3) and thyrotracheoesophageal (group 4) allotransplantation. Allograft revascularization included (1) interposition of donor right subclavian artery--incorporating the inferior thyroid artery--to recipient right carotid artery (end-to-end fashion) and (2) end-to-side anastomosis of donor anterior vena cava to recipient right external jugular vein. All thyrotracheoesophageal blocks were harvested after inferior thyroid artery perfusion with 4 degrees C Euro-Collins solution. The overall lengths of tracheal and esophageal grafts were 10.7 +/- 2.7 cm and 13.4 +/- 3.6 cm, respectively. In the heterotopic groups, all allografts were viable and histologically normal at postmortem examination and the incidence and severity of airway ischemia and rejections (at equal residual levels of cyclosporine) were not different between groups 1 and 2. In the orthotopic groups, the first two pigs died of airway collapse with histologically normal grafts. In the remaining pigs, temporary airway stenting was inserted and allografts remained viable and histologically intact for their entire length 30 days after transplantation. Transplanted tracheal smooth muscles had concentration-dependent contractions and relaxations similar to those of nontransplanted (native) tracheas. This study documents the feasibility of allotransplanting long tracheal and esophageal segments on their own vascular pedicles and demonstrates that allograft preservation and thyrotracheoesophageal transplantation are equally effective in minimizing airway ischemia. Thyrotracheoesophageal transplantation does not enhance recipient alloimmune response compared with thyrotracheal transplantation alone. PMID- 7475133 TI - Constitutive nitric oxide release is impaired after ischemia and reperfusion. AB - Myocardial ischemia and reperfusion may result in endothelial dysfunction and reduced release of nitric oxide. With the use of an amperometric sensor, the first direct measurements of constitutive nitric oxide release from a beating heart were measured from the coronary effluent of isolated working rat hearts subjected to ischemia and reperfusion. Rats, six to eight per group, were randomly studied as follows: control (no pretreatment) and pretreatment with the nitric oxide donor L-arginine (3 mmol/L), its enantiomer D-arginine (3 mmol/L), nitric oxide inhibitor N omega-nitro-L-arginine methyl ester (100 mumol/L), and combined N omega-nitro-L-arginine methyl ester/L-arginine. Isolated hearts were pretreated for 10 minutes before 30 minutes of global ischemia and 30 minutes of reperfusion. A nonischemic control group (n = 4) was continuously perfused with oxygenated unsupplemented buffer. After ischemia/reperfusion, hearts supplemented with L-arginine recovered significantly (p < 0.05) increased developed pressure, first derivative of the aortic pressure (dP/dtmax), and aortic flow compared with all other hearts that underwent ischemia/reperfusion. In addition, nitric oxide release was significantly (p < 0.05) increased during reperfusion in the L arginine group. During reperfusion, the recovery of aortic flow correlated with nitric oxide release (r = 0.81, p < 0.0001). We conclude that after ischemia/reperfusion, endothelial dysfunction results in decreased nitric oxide release, which can be ameliorated with L-arginine pretreatment. The direct cytoprotective properties of nitric oxide may contribute to improved functional recovery in hearts pretreated with L-arginine. Augmentation of the L arginine/nitric oxide pathway may provide a new approach for improved recovery after cardiovascular operations. PMID- 7475134 TI - Simultaneous manipulation of the nitric oxide and prostanoid pathways reduces myocardial reperfusion injury. AB - The effects of aspirin and L-arginine (biological precursor of nitric oxide) on the production of hydroxyl radicals, cyclic guanosine monophosphate levels, vascular tone, and the recovery of the ischemic myocardium were investigated in isolated rat hearts subjected to ischemia and reperfusion. After 30 minutes of perfusion, hearts were arrested with St. Thomas' Hospital cardioplegic solution, global ischemia was induced at 37 degrees C for 45 minutes, and the hearts were then reperfused at 37 degrees C for 30 minutes. The percent change in recovery of pulse pressure and maximal change of this pressure with time were better in the group perfused with Krebs-Henseleit solution containing aspirin plus L-arginine (17% +/- 23%, p = 0.001, and 10% +/- 25%, p = 0.002, respectively) compared with these values in the control group perfused with Krebs-Henseleit solution alone ( 7% +/- 14% and -11% +/- 16%, respectively). Coronary vascular resistance before and after ischemia were lower in the aspirin plus L-arginine group (0.19 +/- 0.03 dynes.sec/cm5, p = 0.001, and 0.23 +/- 0.04 dynes.sec/cm5, p = 0.01, respectively) compared with those of the control group (0.24 +/- 0.02 and 0.28 +/ 0.07 dynes.sec/cm5, respectively). Cyclic guanosine monophosphate levels increased from 22.5 +/- 6 pmol/100 mg of tissue in the control group to 37.1 +/- 8.9 pmol/100 mg (p = 0.002) in the aspirin plus L-arginine group. Adding N omega nitro-L-arginine methyl ester to the perfusion medium caused a deterioration in pulse pressure and maximal change of this pressure with time, a decrease in cyclic guanosine monophosphate, and a rise in coronary vascular resistance. The addition of L-arginine to the solution in the Krebs-Henseleit solution plus aspirin group increased the production of hydroxyl radicals from 0.32 +/- 0.18 nmol/gm per 3 minutes to 0.75 +/- 0.33 nmol/gm per 3 minutes (p = 0.03). Despite the association of nitric oxide with increased hydroxyl radical production, it appears that nitric oxide has an overall beneficial effect on the recovery of the ischemic myocardium. The synergism between aspirin and arginine may be caused in part by the scavenging of hydroxyl radicals. Alternatively, by inhibiting the prostaglandin pathway, aspirin may reduce the generation of superoxide anion, a free radical that inactivates nitric oxide. The prolonged half-life of nitric oxide may explain the increased levels of cyclic guanosine monophosphate seen in the group perfused with Krebs-Henseleit solution plus aspirin plus L-arginine. Aspirin and L-arginine, both readily available, may be useful adjuncts to clinical cardioplegia strategy. PMID- 7475135 TI - Protective effect of nicorandil as an additive to the solution for continuous warm cardioplegia. AB - Experiments were designed to assess whether (1) nicorandil given before global low-flow ischemia or (2) included in low-flow continuous cardioplegia improved the recovery of cardiac function in the isolated rat heart. The first investigated the effect of nicorandil (2, 10, or 100 mumol/L), given for 3 minutes before 30 minutes of normothermic global ischemia, on recovery after 30 minutes of reperfusion. In aerobically perfused hearts, doses of 10 and 100 mumol/L significantly increased coronary flow; the dose of 100 mumol/L exerted a negative inotropic effect. These doses shortened the time to contractile arrest (282 +/- 18 and 276 +/- 22 seconds versus 354 +/- 16 seconds in the control hearts with unmodified ischemia; p < 0.05 in both instances). Nicorandil also improved the postischemic recovery of coronary flow (79.1% +/- 1.7% and 78.0% +/- 1.6%, respectively, versus 71% +/- 1.8%; p < 0.05). However, there was no significant improvement in recovery of contractile function, creatine kinase leakage, or tissue adenosine triphosphate and creatine phosphate contents. Second, pretreatment with nicorandil (10 mumol/L) was shown to increase susceptibility of the hearts to reperfusion-induced ventricular fibrillation from 0% (n = 8) in control hearts to 50% in the drug-treated group (p < 0.05). Third, nicorandil (10 mumol/L) was added to cardioplegic and noncardioplegic solutions infused into the coronary tree throughout 100 minutes of low-flow (0.7 ml/min) ischemia: in eight of nine control hearts electrical activity was maintained throughout, whereas in all nicorandil-treated hearts electrical activity was suppressed for at least part of the time. Nicorandil also reduced the prevalence of ischemic contracture to 0% during continuous infusion of cardioplegic solution (compared with 30% in nicorandil-free control hearts) and improved the recovery of contractile function after 40 minutes of reperfusion. Thus, in the noncardioplegia groups, left ventricular developed pressure recovered to 77.8% +/ 4.0% versus 51.7% +/- 2.6% in control hearts (p < 0.05) and in the cardioplegia groups to 96.2% +/- 4.2% versus 79.7% +/- 5.5% (p < 0.05). Ventricular compliance (the ventricular volume required to achieve a left ventricular end-diastolic pressure of 4 mm Hg) was better preserved in the nicorandil-containing noncardioplegia group (133 +/- 6 microliters) than in the control group (88 +/- 10 microliters; p < 0.05). In conclusion, nicorandil has been shown to (1) reduce ischemic contracture, (2) lessen the effects of ischemic arrest, and (3) improve the postischemic recovery of contractile function. In this species and preparation it may, however, enhance vulnerability to reperfusion-induced arrhythmias. PMID- 7475136 TI - Adenosine triphosphate-sensitive K+ channels mediate postcardioplegia coronary hyperemia. AB - The purpose of the present study was to examine the role of adenosine triphosphate-sensitive potassium channels in mediating the coronary hyperemic response after crystalloid cardioplegia. Thirteen pigs were placed on normothermic cardiopulmonary bypass support. Hearts were arrested with cold (4 degrees C) crystalloid ([K+] 25 mmol/L) cardioplegic solution for 60 minutes. In seven of these pigs, hearts were then reperfused for 60 minutes with warm blood, and the animal was separated from cardiopulmonary bypass. The in vivo responses to the intracoronary administration of the K+ adenosine triphosphate channel blocker glibenclamide (50 gm/kg per minute) or the K+ adenosine triphosphate channel opener pinacidil (2 gm/kg per minute) were evaluated before cardiopulmonary bypass (baseline) and after 2 minutes and 60 minutes of reperfusion in the cardioplegia-reperfusion group. Under baseline conditions, glibenclamide and pinacidil induced a respective decrease and increase in coronary blood flow and an increase and a decrease in coronary vascular resistance. Coronary responses to glibenclamide and pinacidil were markedly enhanced after 2 minutes or 60 minutes of postcardioplegia reperfusion. In vitro responses of coronary arterioles (90 to 180 microns) were examined in a pressurized, no-flow state with video microscopy. The contractile response of coronary arterioles to glibenclamide and the relaxation response to pinacidil were significantly enhanced 2 minutes or 60 minutes after reperfusion (all p < 0.05 versus control). The response to pinacidil was markedly inhibited by glibenclamide, which confirms these antagonistic effects on K+ adenosine triphosphate channels. Decreased tissue concentrations of adenosine triphosphate in the coronary arterial smooth muscle and myocardium were observed after cardioplegia and persisted for up to 60 minutes of reperfusion (both p < 0.05 versus control). These results suggest that coronary hyperemia associated with postischemic cardioplegia is mediated in part by activation of K+ adenosine triphosphate channels in the coronary microcirculation. PMID- 7475138 TI - Acadesine: a new drug that may improve myocardial protection in coronary artery bypass grafting. Results of the first international multicenter study. Multinational Acadesine Study Group. AB - The effect of acadesine, an adenosine-regulating agent, on the incidence of myocardial infarction, all adverse cardiovascular outcomes (myocardial infarction, cardiac death, left ventricular dysfunction, life-threatening arrhythmia, or cerebrovascular accident) and mortality was assessed in 821 patients undergoing coronary artery bypass grafting. Patients were prospectively stratified to a high-risk group (age > 70 years, unstable angina, previous coronary bypass, unsuccessful angioplasty, or ejection fraction < 30%) or a non high-risk group. They were randomized in a double-blind manner to placebo (n = 418) or acadesine (n = 403) by intravenous infusion over 7 hours (0.1 mg/kg per minute) and in the cardioplegic solution (placebo or acadesine; 5 micrograms/ml). Acadesine did not significantly affect the incidence of myocardial infarction in the overall study population, but it significantly reduced the incidence of Q wave myocardial infarction in high-risk patients (placebo, 19.7%; acadesine, 10.0%; p = 0.032). The incidences of all adverse cardiovascular outcomes (placebo, 19.4%; acadesine, 18.4%) and overall mortality (placebo, 3.4%; acadesine, 2.7%) were similar between the two treatment groups. However, acadesine reduced the incidence of cardiac related events that contributed to deaths occurring during the first 3 postoperative days so that the incidence of death in this period was lower (placebo, 1.9%; acadesine, 0.2%; p = 0.038). No adverse events were related to acadesine treatment. Although overall there were no statistically significant between-group differences for the primary study end points, a secondary analysis in a prospectively defined high-risk subgroup suggests that acadesine may be beneficial in some patients. PMID- 7475137 TI - Hyperpolarized cardiac arrest with a potassium-channel opener, aprikalim. AB - Cardioplegic solutions that arrest the heart at or near the resting membrane potential may provide better myocardial protection than standard depolarizing hyperkalemic cardioplegia by reducing both metabolic demand and harmful transmembrane ion fluxes. This hypothesis was investigated in an isolated, blood perfused, rabbit heart Langendorff model during 30 minutes of normothermic global ischemia. Hyperpolarized cardiac arrest induced by aprikalim, an opener of adenosine triphosphate-dependent potassium channels, was compared with hyperkalemic depolarized arrest and with unprotected global ischemia. Left ventricular pressure was recorded over a wide range of balloon volumes before ischemia and 30 minutes after reperfusion. End-diastolic pressure versus balloon volume data were fitted to a two-coefficient exponential relationship. Changes in the diastolic compliance of the left ventricle were assessed by comparison of preischemic and postischemic coefficients within each cardioplegia group. Postischemic recovery of developed pressure was used to assess changes in left ventricular systolic function. The tissue water content of each heart was also determined. Myocardial protection with aprikalim resulted in better postischemic recovery of developed pressure (90% +/- 9%) than either protection with hyperkalemic cardioplegia (73% +/- 11%) or no protection (62% +/- 9%). Myocardial tissue water content in hearts protected with hyperkalemic cardioplegia (77.4% +/ 1.4%) was less than the tissue water content of either unprotected hearts (79.4% +/- 1.2%) or hearts protected with aprikalim (78.7% +/- 0.9%). Despite these differences, neither hyperkalemic cardioplegia (p = 0.15) nor aprikalim cardioplegia (p = 0.30) was associated with a significant postischemic decrease in ventricular compliance. By contrast, unprotected global ischemia was associated with a significant decrease in ventricular compliance (p < 0.001). PMID- 7475140 TI - Elective pneumonectomy for benign lung disease: modern-day mortality and morbidity. AB - This retrospective study of elective pneumonectomy for complicated inflammatory lung disease was done to define modern-day mortality and morbidity. One hundred twenty-four patients received elective pneumonectomy. Patient ages ranged from 6 months to 71 years. Past, recurrent, or new pulmonary tuberculosis was present in 107 patients (86.3%). Clinical presentation involved recurrent infections or severe suppurative sequelae (abscess, empyema). Forty-seven patients had chronic hemoptysis and 25 patients had past or recent massive hemoptysis (> 600 ml of hemoptysis fluid within 24 hours). Nutritional deficiencies were common. One hundred six patients (85.5%) had end-stage destroyed lungs. Evaluative bronchoscopy showed inflammatory endobronchial changes in 106 patients (85.5%), bronchial strictures in 4, and indolent endobronchial tumor in 2. Lung separation was by double-lumen tube in 96 patients, single lung-single tube in 6, bronchus blocker in 6, and prone posture in 9. Extrapleural pneumonectomy was done in 83 patients (66.9%). Fifty-seven of these procedures were left sided and 26 were right sided. Standard transpleural pneumonectomy was done in 41 patients (33.1%): 30 left sided and 11 right sided. Nine pneumonectomies were conducted with the patient in the prone position. Four patients had completion pneumonectomy. Hospital mortality was three deaths (2.4%). Morbidity included postpneumonectomy empyema in 19 patients (15.3%). Seven postoperative bronchopleural fistulas occurred. Empyema in most patients was managed by open pleural drainage (thoracostoma) and later space closure. Pneumonectomy proved effective therapy with low mortality but postpneumonectomy empyema posed serious morbidity. PMID- 7475139 TI - Aprotinin versus desmopressin for patients undergoing operations with cardiopulmonary bypass. A double-blind placebo-controlled study. AB - BACKGROUND: Aprotinin reduces blood loss in operations done with cardiopulmonary bypass, whereas the use of desmopressin remains controversial. We compared aprotinin, desmopressin, and placebo in a double-blind, randomized trial to evaluate bleeding and transfusion requirements. METHODS AND RESULTS: One hundred forty-nine patients (48 received aprotinin, 50 desmopressin, 51 placebo) were included. Blood loss and transfusion requirements were recorded and levels of Factor VIII coagulant activity, von Willebrand's factor, thrombin-antithrombin complexes, and D-dimer were measured. Overall blood loss was 195 +/- 146 ml/m2 in the aprotinin group, 400 +/- 192 ml/m2 in the desmopressin group, and 489 +/- 361 ml/m2 in the placebo group (95% confidence intervals: difference between desmopressin and aprotinin 98 to 312 ml/m2, p < 0.001; difference between placebo and aprotinin 190 to 398 ml/m2, p < 0.001). Twenty-six percent of patients treated with aprotinin, 66% of those treated with desmopressin, and 56% of those treated with placebo were given transfusion (95% confidence intervals: difference between aprotinin versus placebo plus desmopressin 51% to 71%, p < 0.001). Fibrinolytic activation throughout cardiopulmonary bypass was markedly higher with placebo or desmopressin administration. D-dimer level correlated with overall blood loss in patients receiving desmopressin or placebo, but not in those receiving aprotinin. CONCLUSION: Aprotinin administration reduces blood loss and transfusion requirements in cardiopulmonary bypass. This benefit may be explained by a lower activation of fibrinolysis. PMID- 7475141 TI - Completion pneumonectomy: indications and outcome. AB - Completion pneumonectomy has been associated with higher rates of morbidity and mortality and this is reflected in the selection of cases and the indications for the procedure. During a period of 14 years from January 1980 to November 1993, 38 completion pneumonectomies were done by our surgical team, representing 5.1% of all pneumonectomies. There were 24 right and 14 left completion pneumonectomies done in 26 male and 12 female patients with an average age of 61 years (range from 29 to 77 years). Lung malignancy accounted for 26 of these cases in which the indication included local recurrence in 10, second primary tumor in 9, malignancy that developed after resection for benign disease in 2, and pulmonary metastasectomy in 5 cases. Benign diseases were the indication in 12 cases: tuberculosis in 4, bronchiectasis in 4, aspergillosis in 1, and postoperative complications in 3. Additional surgical procedures were necessary in 7 cases: chest wall resection with insertion of prosthesis in 3, thoracoplasty in 2, and omental flap in 2. There was 1 early postoperative death after 5 weeks from adult respiratory distress syndrome. There was no occurrence of bronchopleural fistula, and the 18% associated morbidity rate was a result of bleeding necessitating reexploration in 3 cases, prolonged ventilation in 2, and chronic empyema in 2. Six of these complications (86%) occurred in the group with benign disease. Completion pneumonectomy can be done with an acceptable morbidity in selected patients. Careful technique is important to secure hemostasis and to avoid fistulas. The complication rate is higher when infective disease is involved. PMID- 7475142 TI - Metastatic pattern in adenocarcinoma of the lung. An autopsy study from a cohort of 137 consecutive patients with complete resection. AB - A cohort of 137 patients with completely resected stage I or II adenocarcinoma of the lung was observed from the time of operation; the metastatic pattern determined at autopsy is described in relation to clinical, histologic, and laboratory variables. The pretreatment variables evaluated were performance status, age, gender, lactate dehydrogenase, stage, degree of differentiation, and histologic subtype of adenocarcinoma of the lung. Patients who survived longer than 30 days after operation were eligible for analysis, and 35 autopsies were performed in this patient group (autopsy rate: 39.8%). The most common intrathoracic metastatic sites were mediastinal lymph nodes (43%), lung (31%), pleura (20%), pericardium (9%), and heart (6%). The most common extrathoracic sites were liver (37%), brain (33%), bones (21%), adrenals (17%), and kidneys (17%). Patients undergoing resection for stage I disease had significantly fewer intrathoracic metastases than patients with stage II disease (p = 0.01). Patients who survived less than 1 year had significantly more extrathoracic metastases than patients who survived for a longer period (p = 0.01). Patients with highly differentiated tumors had fewer extrathoracic metastases than patients with less differentiated tumors. No other statistically significant differences were observed. Overall, patients with stage I adenocarcinoma of the lung had better local control of the disease at autopsy than those with stage II disease, but distant metastases are a large problem despite the favorable prognosis of this patient group. The extrathoracic metastatic potential was greatest for less differentiated tumors. An active adjuvant systemic therapy after resection is needed in selected patients with poorly differentiated adenocarcinomas of the lung, even in those with stage I disease. PMID- 7475143 TI - Traumatic aortopulmonary communication: clinicopathologic correlation. PMID- 7475144 TI - Bronchopleural fistula after pneumonectomy: repair and prevention with a deepithelialized latissimus dorsi myocutaneous island flap. PMID- 7475145 TI - Bronchopleural fistula and mechanical ventilation: a simple technique of management? PMID- 7475146 TI - Acquired aortopulmonary fistula in pseudoaneurysm of the aorta six years after a Bentall operation. PMID- 7475147 TI - Balloon-expandable metallic stents in the management of tracheomalacia in neonates. PMID- 7475148 TI - Truncus repair with a valveless conduit in neonates. PMID- 7475149 TI - Biventricular repair in patients with left isomerism. PMID- 7475150 TI - Microembolization during coronary artery bypass grafting. PMID- 7475152 TI - Aorta-coronary bypass in patients with coronary artery disease who do not have angina: a brief follow-up fifteen years after the last case reported. PMID- 7475151 TI - The scientific development of dynamic cardiomyoplasty. PMID- 7475153 TI - Cardiac status and health-related quality of life in the long term after surgical repair of tetralogy of Fallot in infancy and childhood. AB - The long-term results of surgical repair of tetralogy of Fallot were assessed by means of extensive cardiologic examination of 77 nonselected patients 14.7 +/- 2.9 years after surgical repair of tetralogy of Fallot in infancy and childhood. Because of the frequent use of a transannular patch (56%) for the relief of right ventricular outflow tract obstruction, the prevalence of elevated right ventricular systolic pressure was low (8%), but the prevalence of substantial right ventricular dilation with severe pulmonary regurgitation was high (58%). The exercise capacity of patients with a substantially dilated right ventricle proved to be significantly decreased (83% +/- 19% of predicted) when compared with that of patients with a near normal sized right ventricle (96% +/- 13%). Eight out of 10 patients who had needed treatment for symptomatic arrhythmia had supraventricular arrhythmia, which makes supraventricular arrhythmia--in numbers- a more important sequela in the long-term survivors than ventricular arrhythmia. Older age at the time of the operation and longer duration of follow-up were not associated with an increase in prevalence or clinical significance of sequelae. PMID- 7475154 TI - Reoperation after the arterial switch operation for transposition of the great arteries. AB - Although most children after an arterial switch operation for transposition of the great arteries have normal development and cardiac function, a few require reoperation. During the last 10 years, 68 of 753 patients who underwent arterial switch operations (9.3%) underwent 75 reoperations. Thirty underwent early reoperation (< 30 days or during the same hospital stay) and 38 underwent late reoperation. Causes for reoperation included pacemaker insertion (n = 5), left diaphragm plication (n = 4), revision for hemostasis (n = 1), mediastinitis (n = 2), superior vena cava thrombosis (n = 9), subvalvular pulmonic stenosis (n = 5), supravalvular pulmonic stenosis (n = 16), residual atrial (n = 2) or ventricular (n = 8) septal defects, isolated mitral valve insufficiency (n = 2), aortic valve insufficiency (either isolated [n = 1] or in association with mitral incompetence [n = 1] or stenosis [n = 1]), left coronary artery ostial stenosis (n = 1), and recurrent aortic (n = 6) or neoaortic (n = 4) aortic coarctation. In all but 27 patients, the residual defects were already present immediately after the completion of the arterial switch operation; however, only patients with critical lesions were reoperated on early. Interventional catheterization procedures were performed when indicated; however, they only postponed inevitable reoperation. Successful relief of superior vena cava thrombosis was achieved by atriojugular bypass grafting in two patients, by early open thrombectomy in six patients, and by direct patch angioplasty of the superior vena cava once. Patch plasty for subvalvular or supravalvular pulmonic stenosis was carried out in 21 patients, septal defect closure was carried out in nine patients, and pulmonary artery banding was performed in one patient with criss-cross atrioventricular relationship and multiple ventricular septal defects. Valve repair was performed in all five patients with either isolated or combined aortic and mitral valve dysfunction. One patient with left coronary ostial stenosis underwent a patch enlargement of this ostium. Recoarctation was repaired by end-to-end anastomosis in eight patients and by a subclavian flap and a patch angioplasty in one patient each. Seven patients underwent a second reoperation for supravalvular pulmonary stenosis (n = 3), mitral valve replacement (n = 1), ventricular septal defect closure (n = 1), and recurrent coarctation (n = 2). There were six intraoperative (8.8%) and two late deaths. All early deaths occurred after early reoperations.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 7475155 TI - Elevated arginine vasopressin and lowered atrial natriuretic factor associated with hypertension in coarctation of the aorta. AB - Impairment of humoral and neural regulation of blood pressure may contribute to preoperative and postoperative hypertension in coarctation of the aorta and may also affect the release of vasopressin and atrial natriuretic factor. Because vasopressin and atrial natriuretic factor have potent vasoactive effects, we measured plasma vasopressin and atrial natriuretic factor levels by radioimmunoassay before operation and for 5 days after operation in 11 patients aged 9 months to 12 years undergoing coarctation repair and in 12 control patients undergoing other cardiovascular operations. Six patients in the coarctation group required minimal antihypertensive therapy (group I) and five required prolonged intravenous antihypertensive therapy (group II). Before operation, vasopressin levels correlated with systolic blood pressure for all patients in the coarctation group (r = 0.83, p < 0.01) whereas atrial natriuretic factor levels did not. Before operation, atrial natriuretic factor levels were lower (28 +/- 5 vs 41 +/- 7 and 50 +/- 8 pg/ml, p < 0.05) and vasopressin levels were higher (28 +/- 6 vs 5.4 +/- 0.9 and 7 +/- 3 pg/ml, p < 0.05) in group II than in group I or control patients. Vasopressin levels were higher (p < 0.05) on the day of operation and on postoperative days 2 through 5 in group II than in group I and in control patients. Atrial natriuretic factor levels were lower during the day of operation in group II than in group I or in control patients (26 +/- 7 vs 51 +/- 16 and 50 +/- 7 pg/ml, p < 0.05) and remained lower than control values on postoperative days 1 and 3 through 5. Elevated vasopressin and lowered atrial natriuretic factor levels may contribute to preoperative and postoperative hypertension in coarctation. PMID- 7475156 TI - Modified surgical techniques for relief of aortic obstruction in [S,L,L] hearts with rudimentary right ventricle and restrictive bulboventricular foramen. AB - Modified techniques of aortopulmonary anastomosis were performed in six neonates with atrioventricular and ventriculoarterial discordance [S,L,L], double-inlet left ventricle, and restrictive bulboventricular foramen area (mean index 1.10 cm2/m2) with unobstructed aortic arch (n = 3) or with hypoplasia (n = 2) or interruption (n = 1) of the aortic arch. In cases of unobstructed aortic arch, a flap of autogenous aortic tissue was used to augment the posterior aspect of the anastomosis of the main pulmonary artery to the ascending aorta, thus creating the potential for anastomotic growth; this technique is applicable regardless of the position of the ascending aorta relative to the main pulmonary artery. In case of levo-transposition of the aorta with hypoplasia or interruption of the aortic arch, a modified Norwood procedure was performed, in that the proximal ascending aorta was divided at the same level as the main pulmonary artery with subsequent homograft patch augmentation from the main pulmonary artery-ascending aorta anastomosis to the level of the proximal descending aorta; this technique avoids a spiraling incision of the aorta and therefore reduces the risk of torsion of the aortic root with its inherent risks of obstruction of the coronary circulation and aortic or pulmonary valve regurgitation. There was no early or late mortality. At a mean follow-up of 16 months, in all patients, there was unobstructed aortic outflow, as evidenced by echocardiographic absence of a significant ventricular-aortic systolic gradient (mean 4.5 +/- 4 mm Hg) and absence of distal aortic arch obstruction. There was no evidence of aortic or pulmonary valve regurgitation. The reported modified techniques provide effective relief of restrictive bulboventricular foramen and aortic obstruction in [S,L,L] hearts. PMID- 7475157 TI - Surgical repair in children with the Budd-Chiari syndrome. AB - Membranous obstruction of the inferior vena cava at the level of the diaphragm is a rare cause of Budd-Chiari syndrome in children. Medical therapy usually fails. Surgical intervention aims at reestablishing patency of the inferior vena cava and hepatic venous outflow to the right atrium. We report on the management of this condition in 19 children of whom 7 were treated surgically. Indications for operation were persistent ascites, deteriorating liver function, and hepatic and inferior vena caval obstruction without significant collateral circulation. Three pathologic types were identified by ultrasonography and cavography and were confirmed at operation. These were type I (4 cases), with a thin membrane occluding the inferior vena cava at the level of the diaphragm; type II (12 cases), with segmental fibrotic obstruction of the inferior vena cava with variable involvement of hepatic veins; and type III (3 cases), with complete absence or nonvisualization of the inferior vena cava. All procedures were done with an extended midline sternotomy incision, cardiopulmonary bypass, core cooling to 16 degrees to 20 degrees C, and periods of circulatory arrest. Type I lesions necessitated membranectomy; type II lesions necessitated transcaval resection of the occluded confluence of the inferior vena cava and the hepatic vein with repair of the defect with an autogenous pericardial patch. One type II lesion, in addition, called for use of a 14 cm polytetrafluoroethylene tube graft to restore inferior vena caval flow. After the operation, marked clinical improvement was observed with an immediate reduction in liver and spleen size and resolution of ascites. Repeat cavography 10 to 30 days after the operation revealed complete patency in four cases and residual stenosis, which required transiliac balloon angioplasty to normalize the inferior vena cava/right atrial pressure gradient, in 3 cases. Thus eventual relief of hepatic venous outflow obstruction and inferior vena caval flow was restored in all cases. We advocate transcardiac membranectomy and pericardial patch grafting for symptomatic and deteriorating membranous obstruction of the inferior vena cava in children. PMID- 7475159 TI - Cerebral blood flow velocity in pediatric patients is reduced after cardiopulmonary bypass with profound hypothermia. AB - Transcranial Doppler sonography of the middle cerebral artery was used to determine whether cerebral perfusion was detectable in low flow states during operations with cardiopulmonary bypass in pediatric patients. Quantitative and qualitative differences in cerebral blood flow velocity after rewarming in patients treated with continuous low-flow bypass or deep hypothermic circulatory arrest were assessed. To determine whether the alterations in cerebrovascular resistance pattern observed in our patients undergoing profound hypothermia was more a function of perfusion technique than of minimum temperature during operation, a third group of patients treated with moderate hypothermia was studied. The three patient groups were the arrest group (N = 16), comprised of patients undergoing circulatory arrest at 18 degrees to 20 degrees C; the low flow group (N = 16), patients treated with continuous low-flow (cardiac index 0.5 L/min per square meter) bypass at 18 degrees to 20 degrees C; and the moderate group (N = 5), patients treated with moderate hypothermia at 24 degrees to 28 degrees C. Flow velocity was detectable in all patients in the low-flow group, with mean arterial pressures as low as 15 mm Hg. Mean flow velocity was reduced after bypass as compared with prebypass values in both the arrest and low-flow groups (p = 0.0001). Mean flow velocity increased after bypass in the moderate group (p = 0.0001). A Doppler waveform pattern consistent with high cerebrovascular resistance was found in 67% of patients in the arrest group and 44% of those in the low-flow group. None of the patients in the moderate group exhibited such a pattern. Patients treated with profound hypothermia who underwent a period of cold full-flow reperfusion before rewarming did not exhibit this high resistance pattern after rewarming. The present findings indicate that profound hypothermia may evoke changes in the cerebral vasculature that result in decreased mean cerebral blood flow velocity after cardiopulmonary bypass rewarming. A period of cold full-flow reperfusion before rewarming may prevent these alterations and improve cerebral perfusion during rewarming. PMID- 7475158 TI - P-selectin expression in myocardium of children undergoing cardiopulmonary bypass. AB - Cardiopulmonary bypass is a planned support technique that results in a period of myocardial ischemia and reperfusion. In addition, it is associated with an inflammatory response likely involving endothelial cell activation. In previous studies, we showed that E-selectin and intercellular adhesion molecule-1 (ICAM-1) messenger ribonucleic acid (mRNA) are increased in human myocardium after cardiopulmonary bypass. We have now examined the expression of P-selectin mRNA by ribonuclease protection in paired atrial biopsy specimens from 12 patients before and after cardiopulmonary bypass. By means of immunocytochemistry, we have also examined the endothelial cell surface expression of P-selectin protein, as well as that of E-selectin and ICAM-1 in three additional patients. Patient ages ranged from 1 day to 8.5 years (median 12 months), and cardiopulmonary bypass times ranged from 46 to 196 minutes (median 144 minutes). By ribonuclease protection, there was marked variability in the expression of P-selectin in biopsy specimens before bypass. However, when compared with prebypass levels, P selectin mRNA decreased modestly in 10 of 12 patients after bypass (median decrease 1.5-fold, p = 0.016). As seen with immunocytochemistry, P-selectin protein was distributed diffusely through the vascular bed on large vessels and small vessels before bypass but was virtually absent on capillaries in specimens taken after bypass. E-selectin, which was absent in prebypass biopsy specimens, was induced in one of the three specimens after bypass, but no change in ICAM-1 protein expression above baseline was noted. We also find that cultured human endothelial cells treated with tumor necrosis factor-alpha in doses which induce ICAM-1 mRNA simultaneously decrease their expression of P-selectin mRNA as compared with untreated cells. These observations suggest that endothelial P selectin is transcriptionally downregulated after cardiopulmonary bypass at times when E-selectin and ICAM-1 are induced. Furthermore, we find that E-selectin and ICAM-1 are expressed at times and at sites where P-selectin is absent. Although it is possible that P-selectin may have been induced and lost at early times before reperfusion, these data suggest that endothelial P-selectin plays a limited role in the inflammatory response that ensues after cardiopulmonary bypass. PMID- 7475160 TI - Prediction of long-term survival by preoperative exercise testing in patients with depressed ejection fraction undergoing myocardial revascularization. AB - Ejection fraction is a major determinant of morbidity and mortality for patients with ischemic heart disease. Patients with an ejection fraction of 0.40 or less are generally recognized as having a poorer prognosis than those patients with an ejection fraction of 0.50 or better and remain a heterogeneous group. It would be useful if patients with a favorable surgical prognosis could be identified preoperatively. Fifty-five patients who underwent coronary artery bypass grafting and had an ejection fraction less than 0.40 (mean of 0.23 +/- 0.07 standard deviation) were studied by catheter measurement of pulmonary arterial pressure and radionuclide left ventriculography. Heart rate, systemic blood pressure, pulmonary artery pressures, cardiac output, and ejection fraction were measured, at rest, after nitroglycerin was given intravenously and with supine bicycle exercise. Forty-seven patients who had follow-up longer than 4 years were divided into two groups according to their life status (alive or dead) 4 years after operation. Measured variables of exercise stress tests and clinical characteristics were entered into factor analysis to obtain a cardiac function factor score for predicting the life status after 4 years. The cardiac function factor score was highly loaded by stroke index (rest, nitroglycerin), cardiac index (exercise), systemic vascular resistance index (exercise), and history of congestive heart failure. The cardiac function factor provided a predictive value superior to that of any individual variable. By dividing the patients into two groups by cardiac function factor score, the actuarial 5-year survival was 72% versus 17% for good and poor prognosis groups, respectively (p < 0.0001). Preoperative exercise stress testing data integrated by factor analysis provide a predictive tool for patients with a low ejection fraction. PMID- 7475161 TI - High flow demand on small arterial coronary bypass conduits promotes graft spasm. AB - Despite the superior long-term patency of arterial grafts, surgeons are often reluctant to use arterial grafts on coronary vessels that supply large areas of myocardium because postoperative shock may occur. We hypothesized that supramaximal flow through small arterial conduits would decrease distal intraluminal pressure, thereby reducing afterload on the smooth muscle and rendering the arterial graft vulnerable to spasm. Fourteen internal thoracic and eight gastroepiploic arteries were harvested from adult pigs (220 to 250 pounds). Arteries were mounted on a computer-controlled perfusion system with inflow pressure at 80 mm Hg and outflow resistance adjusted to simulate normal (in situ) or supramaximal (coronary artery bypass graft) flow demands. Artery pressures and flow rates were measured at baseline and after norepinephrine was added to the system. Internal thoracic arteries had no hemodynamic response to norepinephrine at normal flow. Under supramaximal flow demands, large internal thoracic arteries (2.5 to 3.0 mm) had no hemodynamic response to norepinephrine. However, for small internal thoracic arteries (2.1 to 2.9 mm), norepinephrine reduced distal internal thoracic arterial pressure (63.2 +/- 2.2 to 27.0 +/- 1.9 mm Hg) and flow rate (99.4 +/- 5.0 to 45.4 +/- 2.7 ml/min, median effective dose = 9.12 x 10(-9) mol/L). Under normal flow demands, the flow rate in gastroepiploic arteries (1.0 to 2.0 mm diameter) decreased (14.1 +/- 0.5 to 4.8 +/- 0.8 ml/min, p < 0.05) only at high concentrations of norepinephrine (median effective dose = 1.26 x 10(-6) mol/L). Supramaximal flow demands reduced distal gastroepiploic arterial pressure (77.5 +/- 0.5 to 49.5 +/- 3.8 mm Hg, p < 0.05), which resulted in a greater decrease in flow rate (80.0 +/- 3.7 to 6.8 +/- 1.6 ml/min, p < 0.05) at lower concentrations of norepinephrine, (median effective dose = 3.24 x 10(-8) mol/L, p < 0.05). In four studies in internal thoracic arteries and eight in gastroepiploic arteries, arteries were cut in half, reattached, and reperfused. The proximal half of the internal thoracic artery did not respond to norepinephrine, but the distal half had a 53% +/- 7% decrease in flow. Both gastroepiploic artery halves reacted and flow rate decreased by 88% +/- 2% (proximal half) and 89% +/- 3% (distal half). In conclusion, small arterial conduits develop large transconduit pressure gradients under supramaximal flow demands. Under these conditions, arteries are very sensitive to vasoconstrictors and flow may cease with higher drug concentrations.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 7475162 TI - Recombinant aprotinin in coronary artery bypass graft operations. AB - OBJECTIVE: To evaluate the role of recombinant bovine aprotinin in reducing blood loss in coronary artery bypass graft surgery. DESIGN: An open-label, randomized, controlled study evaluating two dosage levels of recombinant aprotinin. SETTING: Two acute care hospitals (Northwestern Memorial Hospital, Chicago, Ill., and the Scott & White Memorial Hospital, Temple, Texas). PATIENTS: Patients undergoing primary and reoperation coronary artery bypass grafting were assigned to groups by means of a computer-generated table of random numbers. Treated (n = 48) and control (n = 36) patients did not differ significantly in age, sex, weight, number of grafts, or preoperative hemoglobin level. INTERVENTIONS: Recombinant aprotinin was given at two dosages. Dosage level 1 consisted of a bolus of 2 mg/kg intravenously immediately after the induction of anesthesia, 1 mg/kg added to each liter of the oxygenator prime, and 0.5 mg.kg-1.hr-1 infused continuously during operation. At dosage level 2, doses were doubled. Intraoperative monitoring of anti-factor Xa activity was performed, and additional doses of heparin were given on the basis of anti-factor Xa results. MAIN OUTCOME MEASURES: Preoperative and postoperative hemoglobin levels, amounts of autotransfusion device and chest tube drainage blood, and transfusions of allogeneic red blood cells. Adverse clinical events (alterations in renal function, graft thrombosis, myocardial infarction, and death) were recorded. RESULTS: Additional heparin was given to 48% patients in the aprotinin group and to 44% of control patients. Overall red blood cell loss (in milliliters, mean +/- standard deviation [SD]) was decreased with aprotinin at dosage level 1 for reoperations (1040 +/- 162 vs 1544 +/- 198, p < 0.01), and at dosage level 2 for all operations (primary operations, 886 +/- 362 vs 1333 +/- 618, p = 0.02; reoperations, 1191 +/- 560 vs 1815 +/- 1116, p = 0.2). Fewer patients in the aprotinin than in the control group had transfusions of donated blood (6/48 vs 12/36, p = 0.02) or reinfusion of chest tube drainage blood (12/48 vs 20/36, p < 0.01). Among patients receiving dosage level 1, there were no myocardial infarctions or deaths. At dosage level 2, one patient had profound bradycardia and died on day 12 and two patients had late graft closures. Two control patients had hypotension after bypass necessitating intraaortic balloon pumps, and one of these patients died. Postoperative increases in blood urea nitrogen and creatinine levels were small in both aprotinin and control groups. No hypersensitivity or other allergic reactions occurred. CONCLUSION: We conclude that, at the dosages given, recombinant bovine aprotinin decreases surgical blood loss and transfusion requirements in patients undergoing coronary artery bypass grafting, but its use requires appropriate monitoring of heparin use during bypass. Whether higher dosages of aprotinin increase the risk of graft thrombosis must be further assessed with a larger patient sample. PMID- 7475163 TI - Determinants of operative mortality in reoperative coronary artery bypass grafting. AB - Previously suggested risk factors for operative mortality in reoperative coronary artery bypass grafting are contradictory. Therefore, we analyzed our data of 622 patients who underwent reoperative bypass grafting from January 1986 through June 1993. Among these patients, 258 had saphenous vein grafts alone and 364 had internal mammary artery grafting, including unilateral (342 patients) and bilateral (22 patients) mammary artery grafting with or without additional saphenous vein grafting. Overall operative mortality was 11.4% for reoperation compared with only 3.6% for primary bypass grafting during the same time frame. To determine risk factors for mortality and the influence of internal mammary artery grafting on the outcome, we analyzed 82 variables (31 preoperative, 17 intraoperative, and 34 postoperative) by univariate analysis. Significant variables or the variables having a trend (p < 0.2) to be associated with the mortality were included in stepwise multiple logistic regression analyses. Two regression analyses were separately performed. Regression 1 only included preoperative and intraoperative variables whereas regression 2 included postoperative variables as well. The logistic regressions demonstrate that preoperative variables (low ejection fraction [p = 0.0002], old age [p = 0.003], female gender [p = 0.011], and history of arrhythmia [p = 0.023]), intraoperative variables (emergency operation [p = 0.0001] and long perfusion time [p = 0.0001]), and postoperative variables (complications) are independently associated with higher mortality. Unlike previously described results, aortic crossclamp time, route of cardioplegia, use of internal mammary artery, number of grafts, and year of operation are not associated with operative mortality. The identification of these risk factors may have important implications in further improvement of the results of reoperative coronary artery bypass grafting. PMID- 7475164 TI - Coronary artery bypass without cardiopulmonary bypass: analysis of short-term and mid-term outcome in 220 patients. AB - Two hundred twenty patients, preferentially those with high-risk conditions, underwent coronary artery bypass grafting without cardiopulmonary bypass. Early unfavorable outcome events included operative mortality (7 patients, 3.2%), nonfatal perioperative myocardial infarction (6 patients, 2.7%), cerebrovascular accident (1 patient, 0.4%), and sternal infection (3 patients, 1.4%). There were two deaths (13%) among 15 patients with calcified aorta and four (12%) in 33 patients who underwent emergency operation. Multivariate analysis revealed these two risk factors to be the only predictors of early mortality (odds ratios, 8.0 and 9.8, respectively). Preoperative risk factors such as left ventricular dysfunction (ejection fraction < or = 35%) (40 patients, 18%), congestive heart failure (46 patients, 21%), acute myocardial infarction (59 patients, 27%), cardiogenic shock (7 patients, 3%), age 70 years or older (59 patients, 27%), renal failure (19 patients, 9%), and cerebrovascular accident and carotid disease (11 patients, 5%) were not found to be major predictors of early mortality or unfavorable outcome. During 12 months of follow-up (range 1 to 21 months), there were four cardiac and three noncardiac deaths (1-year actuarial survival 93%) and 17 cases (7.7%) of early return of angina. Calcified aorta, nonuse of the internal mammary artery, reoperation, and diabetes mellitus were independent predictors of unfavorable events. We conclude that coronary artery bypass grafting without cardiopulmonary bypass can be done with relatively low operative mortality, although there seems to be an increased risk for early return of angina. This procedure should therefore be considered for patients with appropriate coronary anatomy, in whom cardiopulmonary bypass poses a high risk. This procedure is still hazardous with calcified aorta or emergency operation. PMID- 7475165 TI - A hybrid vascular model biomimicking the hierarchic structure of arterial wall: neointimal stability and neoarterial regeneration process under arterial circulation. AB - A layered-structure hybrid vascular graft, mimicking the hierarchic structure of the intima and media of a natural artery, is expected to exhibit antithrombogenicity and to accelerate neoarterial tissue formation. Two models of hybrid vascular grafts were prepared on knitted Dacron fabric grafts (inner diameter 4 mm, length 6 cm). Model I grafts consisted of an endothelial cell monolayer formed on collagenous matrix, and model II grafts consisted of an endothelial cell monolayer that formed on hybrid collagenous medial tissue in which smooth muscle cells were incorporated. Both models (n = 17 for each model) were implanted bilaterally in carotid arteries and left in place for up to 26 weeks. Although all of the implanted grafts were patent, the two models significantly differed in the degree of maturity of the regenerated neoarterial wall especially at earlier implantation periods. At 2 weeks, model II grafts showed a much higher degree of neointimal integrity than model I grafts: a smooth and organized neointimal layer was formed, which was free from leukocyte adhesion. On further increase of implantation time, the formation of neomedia (subendothelial smooth muscle cell layers) and circumferential orientation of both smooth muscle cells and collagenous extracellular matrix were much more advanced in model II grafts than in model I grafts. At 26 weeks after implantation, layered elastic laminae regenerated along circumferentially oriented smooth muscle cells in neomedia were observed only in model II grafts. Irrespective of model, little excessive smooth muscle cell proliferation occurred. A hierarchically structured hybrid graft eventually provided a more integrated neointimal layer and accelerated neoarterial tissue formation much more than model I grafts. The significance of the incorporation of smooth muscle cells into hybrid grafts is discussed. PMID- 7475167 TI - Studies of hypoxemic/reoxygenation injury: I. Linkage between cardiac function and oxidant damage. PMID- 7475166 TI - Effects of cryopreservation on the proliferation and anticoagulant activity of human saphenous vein endothelial cells. AB - Human saphenous veins were cryopreserved in 4% human albumin and 10% dimethyl sulfoxide. The effect of cryopreservation on endothelial cells was studied in terms of the anticoagulant activity of thrombomodulin and in terms of cell proliferation. After storage for 2 weeks at -150 degrees C, 0.45 +/- 0.07 x 10(5) endothelial cells/cm2 were detected in cryopreserved veins and 1.03 +/- 0.04 x 10(5) endothelial cells/cm2 in fresh veins (p < 0.01). The thrombin-catalyzed activation of protein C decreased after cryopreservation, indicating altered thrombomodulin activity in the endothelial cells. On a cell number basis, the release of soluble thrombomodulin was three times higher from the cryopreserved endothelium than from the fresh endothelium (p < 0.05). The amount of spontaneous release of von Willebrand factor from the endothelial surface was not significantly different between fresh and cryopreserved veins. Endothelial cells were cultured from fresh veins and from their cryopreserved counterparts. On plating of endothelial cells in primary culture, the number of adhered cells was 0.9 +/- 0.09 x 10(3) cells/cm2 from fresh veins and 0.25 +/- 0.03 x 10(3) cells/cm2 from cryopreserved veins (p < 0.01). The positive immunohistochemical stain for von Willebrand factor indicated that the endothelial cell character was maintained after cryopreservation. The endothelial desquamation with loss of anticoagulant function and the slow proliferation of surviving cells in vitro suggest an impaired endothelial healing in vivo. The loss of anticoagulant activity complicates the problems of the exposure of thrombogenic subendothelial matrix to blood in implanted cryopreserved veins. PMID- 7475168 TI - Studies of hypoxemic/reoxygenation injury: without aortic clamping. II. Evidence for reoxygenation damage. AB - This study tested the hypothesis that the developing heart is susceptible to oxygen-mediated damage after reintroduction of molecular oxygen and that this "unintended" reoxygenation injury causes lipid peroxidation and functional depression that may contribute to perioperative cardiac dysfunction. Among 49 Duroc-Yorkshire piglets (2 to 3 weeks old, 3 to 5 kg) 15 control studies were done without hypoxemia to test the effects of the surgical preparation (n = 10) and 60 minutes of cardiopulmonary bypass (n = 5). Twenty-nine piglets underwent up to 2 hours of ventilator hypoxemia (with inspired oxygen fraction reduced to 6% to 7%) to lower arterial oxygen tension to approximately 25 mm Hg. Five piglets did not undergo reoxygenation to determine alterations caused by hypoxemia alone. Twenty-four others received reoxygenation by either raising ventilator inspired oxygen fraction to 1.0 (n = 12) or instituting cardiopulmonary bypass at oxygen tension 400 mm Hg (n = 12). Ventilator hypoxemia produced sufficient hemodynamic compromise and metabolic acidosis that 18 piglets required premature reoxygenation (78 +/- 12 minutes). To avoid the influence of acidosis and hemodynamic deterioration during ventilator hypoxemia, five others underwent 30 minutes of hypoxemia during cardiopulmonary bypass (circuit primed with blood at oxygen tension 25 mm Hg) and 30 minutes of reoxygenation (oxygen tension 400 mm Hg) during cardiopulmonary bypass. Biochemical markers of oxidant damage included measurement of coronary sinus and myocardial conjugated dienes to determine lipid peroxidation and antioxidant reserve capacity assessed by incubating myocardial tissue in the oxidant t-butylhydroperoxide. Functional recovery was determined by inscribing pressure volume loops to determine end systolic elastance and Starling curves by volume infusion. No biochemical or functional changes occurred in control piglets. Hypoxemia without reoxygenation did not change plasma levels of conjugated dienes, but lowered antioxidant reserve capacity 24%. Reoxygenation by ventilator caused refractory ventricular arrhythmias in two piglets (17% mortality), raised levels of conjugated dienes 45%, and reduced antioxidant reserve capacity 40% with recovery of 39% of mechanical function in the survivors. Comparable biochemical and functional changes occurred in piglets undergoing ventilator hypoxemia and/or cardiopulmonary bypass hypoxemia and reoxygenation on cardiopulmonary bypass. We conclude that hypoxemia increases vulnerability to reoxygenation damage by reducing antioxidant reserve capacity and that reoxygenation by either ventilator or cardiopulmonary bypass produces oxidant damage with resultant functional depression that is not a result of cardiopulmonary bypass. These findings suggest that initiation of cardiopulmonary bypass in cyanotic immature subjects causes an unintended reoxygenation injury, which may increase vulnerability to subsequent ischemia during surgical repair. PMID- 7475169 TI - Studies of hypoxemic/reoxygenation injury: without aortic clamping. III. Comparison of the magnitude of damage by hypoxemia/reoxygenation versus ischemia/reperfusion. AB - The immature heart is more tolerant to ischemia than the adult heart, yet infants with cyanosis show myocardial damage after surgical correction of congenital cardiac defects causing hypoxemia. This study tested the hypothesis that the hypoxemic developing heart is susceptible to oxygen-mediated damage when it is reoxygenated during cardiopulmonary bypass and that this hypoxemic/reoxygenation injury is more severe than ischemic/reperfusion stress. Fifteen Duroc-Yorkshire piglets (2 to 3 weeks old, 3 to 5 kg) underwent 60 minutes of 37 degrees C cardiopulmonary bypass. Five piglets (control) were not made ischemic or hypoxemic. Five underwent 30 minutes of normothermic ischemia (aortic clamping) and 25 minutes of reperfusion before cardiopulmonary bypass was discontinued. Five others underwent 30 minutes of hypoxemia (bypass circuit primed with blood with oxygen tension 20 to 30 mm Hg) and 30 minutes of reoxygenation during cardiopulmonary bypass. Functional (left-ventricular contractility) and biochemical (levels of plasma and tissue conjugated dienes and antioxidant reserve capacity) measurements were made before ischemia/hypoxemia and after reperfusion/reoxygenation. Cardiopulmonary bypass (no ischemia or hypoxemia) caused no changes in left-ventricular function or coronary sinus levels of conjugated dienes. The tolerance to normothermic ischemia was confirmed, inasmuch as left-ventricular function returned to 108% of control values and coronary sinus levels of conjugated dienes did not rise after reperfusion. Conversely, reoxygenation raised plasma levels of conjugated dienes in coronary sinus blood in the hypoxic group 57% compared with end-hypoxic levels (p < 0.05 versus end hypoxic levels and versus ischemia, by analysis of variance). Antioxidant reserve capacity showed the lowest levels (highest production of malondialdehyde) in the hypoxemic group (51% higher than control values; p < 0.05 by analysis of variance). These biochemical changes were associated with a 62% depression of left-ventricular function after bypass because end-systolic elastance recovered only 38% of control levels (p < 0.05 by analysis of variance). These data confirm the tolerance of the immature heart to ischemia and reperfusion and document a hypoxemic/reoxygenation injury that occurs in immature hearts reoxygenated during bypass. Hypoxemia seems to render the developing heart susceptible to reoxygenation damage that depresses postbypass function and is associated with lipid peroxidation. These findings suggest that starting bypass in cyanotic immature subjects causes an unintended reoxygenation injury that may potentially be counteracted by adding antioxidants to the prime of the extracorporeal circuit. PMID- 7475170 TI - Studies of hypoxemic/reoxygenation injury: without aortic clamping. IV. Role of the iron-catalyzed pathway: deferoxamine. AB - This study tests the hypothesis that an iron chelator, deferoxamine, can reduce oxygen-mediated myocardial injury and avoid myocardial dysfunction after cardiopulmonary bypass by its action on the iron-catalyzed Haber-Weiss pathway. Twenty-one immature 2- to 3-week-old piglets were placed on cardiopulmonary bypass for 120 minutes, and five piglets served as biochemical controls without cardiopulmonary bypass. Five piglets underwent cardiopulmonary bypass without hypoxemia (cardiopulmonary bypass control). Sixteen others became hypoxemic while undergoing cardiopulmonary bypass for 60 minutes by lowering oxygen tension to about 25 mm Hg, followed by reoxygenation at oxygen tension about 400 mm Hg for 60 minutes. Oxygen delivery was maintained during hypoxemia by increasing cardiopulmonary bypass flow and hematocrit level. In seven piglets deferoxamine (50 mg/kg total dose) was given both intravenously just before reoxygenation and by a bolus injection (5 mg/kg) into the cardiopulmonary bypass circuit; nine others were not treated (no therapy). Myocardial function after cardiopulmonary bypass was evaluated form end-systolic elastance (conductance catheter) and Starling curve analysis. Myocardial conjugated diene production and creatine kinase leakage were assessed as biochemical markers of injury, and antioxidant reserve capacity was determined by measuring malondialdehyde in postcardiopulmonary bypass myocardium incubated in the oxidant, t butylhydroperoxide. Cardiopulmonary bypass without hypoxemia caused no oxidant or functional damage. Conversely, reoxygenation (no therapy) raised myocardial conjugated diene levels and creatine kinase production (conjugated diene: 3.5 +/- 0.7 absorbance 233 nm/min/100 g, creatine kinase: 8.5 +/- 1.5 U/min/100 g; p < 0.05 versus cardiopulmonary bypass control), reduced antioxidant reserve capacity (malondialdehyde: 1115 +/- 60 nmol/g protein at 4 mmol/L t-butylhydroperoxide; p < 0.05 versus control), and produced severe post-bypass dysfunction (end-systolic elastance recovered only 39% +/- 7%, p < 0.05 versus cardiopulmonary bypass control). Deferoxamine avoided conjugated diene production and creatine kinase release and retained normal antioxidant reserve, and functional recovery was complete (95% +/- 11%, p < 0.05 versus no treatment). These findings show that iron-catalyzed oxidants may contribute to a reoxygenation injury and imply that deferoxamine may be used to surgical advantage. PMID- 7475171 TI - Studies of hypoxemic/reoxygenation injury: without aortic clamping. V. Role of the L-arginine-nitric oxide pathway: the nitric oxide paradox. AB - This study tests the hypothesis that nitric oxide, which is endothelial-derived relaxing factor, produces reoxygenation injury via the L-arginine-nitric oxide pathway in hypoxemic immature hearts when they are placed on cardiopulmonary bypass. Twenty 3-week-old piglets undergoing 2 hours of hypoxemia (oxygen tension about 25 mm Hg) on a ventilator were reoxygenated by initiating cardiopulmonary bypass (oxygen tension about 400 mm Hg). Five animals were not treated, whereas the pump circuit was primed with the nitric oxide-synthase inhibitor NG-nitro-L arginine methyl ester (L-NAME, 4 mg/kg) in five piglets. L-Arginine, the substrate for nitric oxide, was administered in a fivefold excess (20 mg/kg), together with L-NAME in five piglets (L-NAME and L-arginine), and given alone in five other piglets (L-arginine). Five normoxemic, instrumented piglets served as a control group, and five others underwent 30 minutes of cardiopulmonary bypass without preceding hypoxemia. Left ventricular contractility was determined as end systolic elastance by pressure-dimension loops. Myocardial conjugated dienes were measured as a marker of lipid peroxidation, and the antioxidant reserve capacity (malondialdehyde production in tissue incubated with t-butylhydroperoxide) was measured. Nitric oxide level was determined in coronary sinus plasma as its spontaneous oxidation product, nitrite. Cardiopulmonary bypass per se did not alter left ventricular contractility, cause lipid peroxidation, or lower antioxidant capacity. Reoxygenation without treatment depressed cardiac contractility (end-systolic elastance 38% +/- 12% of control*), raised nitric oxide (127% above hypoxemic values), increased conjugated dienes (1.3 +/- 0.2 vs 0.7 +/- 0.1, control*), and reduced antioxidant reserve capacity (910 +/- 59 vs 471 +/- 30, control*). Inhibition of nitric oxide production by L-NAME improved end-systolic elastance to 84% +/- 12%,** limited conjugated diene elution (0.8 +/ 0.1 vs 1.3 +/- 0.2, no treatment**), and improved antioxidant reserve capacity (679 +/- 69 vs 910 +/- 59, no treatment**). Conversely, L-arginine counteracted these beneficial effects of L-NAME, because left ventricular function recovered only 24% +/- 6%,* conjugated dienes were 1.2 +/- 0.1,* and antioxidant reserve capacity was 826 +/- 70.* L-Arginine alone caused the same deleterious biochemical changes as L-NAME/L-arginine and resulted in 60% mortality. The close relationship between postbypass left ventricular dysfunction (percent end systolic elastance) and myocardial conjugated diene production (r = 0.752) provides in vivo evidence that lipid peroxidation contributes to myocardial dysfunction after reoxygenation.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 7475172 TI - Studies of hypoxemic/reoxygenation injury: without aortic clamping. VI. Counteraction of oxidant damage by exogenous antioxidants: N-(2 mercaptopropionyl)-glycine and catalase. AB - This study tests the hypothesis that antioxidants administered before reoxygenation can reduce oxygen-mediated damage and improve myocardial performance. Of 25 Duroc-Yorkshire piglets (2 to 3 weeks, 3 to 5 kg) five underwent 60 minutes of cardiopulmonary bypass without hypoxemia (control group), and five others underwent 30 minutes of hypoxemia on cardiopulmonary bypass with a circuit primed with oxygen tension about 25 mm Hg blood followed by reoxygenation on cardiopulmonary bypass (no treatment). In vitro studies were performed to obtain the optimal dosage of the antioxidants N-(2 mercaptopropionyl)-glycine and and catalase to be used in subsequent in vivo experimental studies; cardiac homogenates were incubated in 0 to 5 mmol/L concentrations of the oxidant t-butylhydroperoxide and malondialdehyde production was measured. Fifteen piglets were made hypoxemic on cardiopulmonary bypass for 30 minutes, and the antioxidants N-(2-mercaptopropionyl)-glycine at either 30 or 80 mg/kg body weight or N-(2-mercaptopropionyl)-glycine, 30 mg/kg body weight, and catalase, 50,000 U/kg body weight, were added to the cardiopulmonary bypass circuit 15 minutes before reoxygenation. Left ventricular contractility, which was expressed as end-systolic elastance, was measured by conductance catheter before hypoxemia and after reoxygenation. Myocardial antioxidant reserve capacity was determined after reoxygenation by incubating cardiac homogenates in the oxidant t-butylhydroperoxide and measuring subsequent malondialdehyde elution. The in vitro bioassay studies showed a dose-dependent reduction of lipid peroxidation with N-(2-mercaptopropionyl)-glycine, with maximal benefits of a 40% decrease and malondialdehyde elaboration occurring with N-(2-mercaptopropionyl) glycine and catalase compared with untreated cardiac homogenates. Cardiopulmonary bypass (no hypoxemia) caused no oxidant damage or changes in contractile function after cardiopulmonary bypass. Reoxygenation without treatment raised conjugated diene levels 57%,* lowered antioxidant reserve capacity 51%,* and was associated with only 38%* recovery of contractile function (p < 0.05 vs control). In contrast, treatment with antioxidants avoided lipid peroxidation, maintained antioxidant reserve capacity, and resulted in a dose-dependent improvement in left ventricular contractility with complete recovery occurring in N-(2 mercaptopropionyl)-glycine and catalase-treated piglets (*p < 0.05 vs no treatment). This study confirms the occurrence of hypoxemic/reoxygenation injury in immature hearts placed on cardiopulmonary bypass and shows that biochemical and functional damage can be counteracted by adding antioxidants to the cardiopulmonary bypass priming fluid. Contractile function improved in a dose dependent manner, and oxygen-mediated damage could be avoided by mercaptopropionyl glycine/catalase treatment.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 7475173 TI - Studies of hypoxemic/reoxygenation injury: without aortic clamping. VII. Counteraction of oxidant damage by exogenous antioxidants: coenzyme Q10. AB - Coenzyme Q10 (CoQ10) is a natural mitochondrial respiratory chain constituent with antioxidant properties. This study tests the hypothesis that CoQ10 administered before the onset of reoxygenation on cardiopulmonary bypass, can reduce oxygen-mediated myocardial injury and avoid myocardial dysfunction after cardiopulmonary bypass. The antioxidant properties of CoQ10 were confirmed by an in vitro study in which normal myocardial homogenates were incubated with the oxidant, t-butylhydroperoxide. Fifteen immature piglets (< 3 weeks old) were placed on 60 minutes of cardiopulmonary bypass. Five piglets underwent cardiopulmonary bypass without hypoxemia (oxygen tension about 400 mm Hg). Ten others became hypoxemic on cardiopulmonary bypass for 30 minutes by lowering oxygen tension to approximately 25 mm Hg, followed by reoxygenation at oxygen tension about 400 mm Hg for 30 minutes. In five piglets, CoQ10 (45 mg/kg) was added to the cardiopulmonary bypass circuit 15 minutes before reoxygenation, and five others were not treated (no treatment). Myocardial function after cardiopulmonary bypass was evaluated from end-systolic elastance (conductance catheter), oxidant damage (lipid peroxidation) was assessed by measuring conjugated diene levels in coronary sinus blood, and antioxidant reserve capacity was determined by measuring malondialdehyde in myocardium after cardiopulmonary bypass incubated in the oxidant, t-butylhydroperoxide. Cardiopulmonary bypass without hypoxemia caused no oxidant damage and allowed complete functional recovery. Reoxygenated hearts (no treatment) showed a progressive increase in conjugated diene levels in coronary sinus blood after reoxygenation (2.3 +/- 0.6 A233 nm/0.5 ml plasma at 30 minutes after reoxygenation) and reduced antioxidant reserve capacity (malondialdehyde: 1219 +/- 157 nmol/g protein at 4.0 mmol/L t butylhydroperoxide), resulting in severe postbypass dysfunction (percent end systolic elastance = 38 +/- 6). Conversely, CoQ10 treatment avoided the increase in conjugated diene levels (2.1 +/- 0.6 vs 1.1 +/- 0.3, p < 0.05 vs no treatment), retained normal antioxidant reserve (896 +/- 76 nmol/g protein, p < 0.05 vs no treatment), and allowed nearly complete recovery of function (94% +/- 7%, p < 0.05 vs no treatment). We conclude that reoxygenation of the hypoxemic immature heart on cardiopulmonary bypass causes oxygen-mediated myocardial injury, which can be limited by CoQ10 treatment before reoxygenation. These findings imply that coenzyme Q10 can be used to surgical advantage in cyanotic patients, because therapeutic blood levels can be achieved by preoperative oral administration of this approved drug. PMID- 7475174 TI - Studies of hypoxemic/reoxygenation injury: without aortic clamping. VIII. Counteraction of oxidant damage by exogenous glutamate and aspartate. AB - Previous studies show that (1) hypoxemia depletes immature myocardium of amino acid substrates and their replenishment improves ischemic tolerance, (2) reoxygenation on cardiopulmonary bypass causes oxygen-mediated damage without added ischemia, and (3) this damage may be related to the nitric oxide-L-arginine pathway that is affected by amino acid metabolism. This study tests the hypothesis that priming the cardiopulmonary bypass circuit with glutamate and aspartate limits reoxygenation damage. Of 22 immature Duroc-Yorkshire piglets (< 3 weeks old), five were observed over a 5-hour period (control), and five others underwent 30 minutes of CPB without hypoxemia (cardiopulmonary bypass control). Twelve others became hypoxemic by reducing ventilator inspired oxygen fraction to 6% to 7% (oxygen tension about 25 mm Hg) before reoxygenation on cardiopulmonary bypass for 30 minutes. Of these five were untreated (no treatment), and the cardiopulmonary bypass circuit was primed with 5 mmol/L glutamate and aspartate in seven others (treatment). Left ventricular function before and after bypass was measured by inscribing pressure-volume loops (end-systolic elastance). Myocardial conjugated diene levels were measured to detect lipid peroxidation, and antioxidant reserve capacity was tested by incubating cardiac muscle with the oxidant t-butylhydroperoxide to determine the susceptibility to subsequent oxidant injury. CPB (no hypoxemia) allowed complete functional recovery without changing conjugated dienes and antioxidant reserve capacity, whereas reoxygenation injury developed in untreated hearts. This was characterized by reduced contractility (elastance end-systolic recovered only 37% +/- 8%*), increased conjugated diene levels (1.3 +/- 0.1 vs 0.7 +/- 0.1*), and decreased antioxidant reserve capacity (910 +/- 59 vs 471 +/- 30 malondialdehyde nmol/g protein at 2 mmol/L t-butylhydroperoxide*). In contrast, priming the cardiopulmonary bypass circuit with glutamate and aspartate resulted in significantly better left ventricular functional recovery (75% +/- 8% vs 37% +/- 8%*), minimal conjugated diene production (0.8 +/- 0.1 vs 1.3 +/- 0.1*), and improved antioxidant reserve capacity (726 +/- 27 vs 910 +/- 59 malondialdehyde nmol/g protein*) (*p < 0.05 vs cardiopulmonary bypass control). We conclude that reoxygenation of immature hypoxemic piglets by the initiation of cardiopulmonary bypass causes myocardial dysfunction, lipid peroxidation, and reduced tolerance to oxidant stress, which may increase vulnerability to subsequent ischemia (i.e., aortic crossclamping). These data suggest that supplementing the prime of cardiopulmonary bypass circuit with glutamate and aspartate may reduce these deleterious consequences of reoxygenation. PMID- 7475175 TI - Studies of hypoxemic/reoxygenation injury: without aortic clamping. IX. Importance of avoiding perioperative hyperoxemia in the setting of previous cyanosis. AB - This study of an in vivo infantile piglet model of compensated hypoxemia tests the hypothesis that reoxygenation on hyperoxemic cardiopulmonary bypass produces oxygen-mediated myocardial injury that can be limited by normoxemic management of cardiopulmonary bypass and the interval after cardiopulmonary bypass. Twenty-five immature piglets (< 3 weeks old) were placed on 120 minutes of cardiopulmonary bypass and five piglets served as a biochemical control group without cardiopulmonary bypass. Five piglets underwent cardiopulmonary bypass without hypoxemia (cardiopulmonary bypass control). Twenty others became hypoxemic on cardiopulmonary bypass for 60 minutes by lowering oxygen tension to about 25 mm Hg. The study was terminated in five piglets at the end of hypoxemia, whereas 15 others were reoxygenated at an oxygen tension about 400 mm Hg or about 100 mm Hg for 60 minutes. Oxygen delivery was maintained during hypoxemia by increasing cardiopulmonary bypass flow and hematocrit level to avoid metabolic acidosis and lactate production. Myocardial function after cardiopulmonary bypass was evaluated from end-systolic elastance (conductance catheter) and Starling curve analysis. Myocardial conjugated diene production and creatine kinase leakage were assessed as biochemical markers of injury, and antioxidant reserve capacity was determined by measuring malondialdehyde after cardiopulmonary bypass in myocardium incubated in the oxidant, t-butylhydroperoxide. Cardiopulmonary bypass without hypoxemia caused no oxidant or functional damage. Conversely, reoxygenation at an oxygen tension about 400 mm Hg raised myocardial conjugated diene level and creatine kinase production (CD: 3.5 +/- 0.7 A233 nm/min/100 g, creatine kinase: 8.5 +/- 1.5 U/min/100 g, p < 0.05 vs cardiopulmonary bypass control), reduced antioxidant reserve capacity (malondialdehyde: 1115 +/- 60 nmol/g protein at 4.0 mmol t-butylhydroperoxide, p < 0.05 vs control), and produced severe postbypass dysfunction (end-systolic elastance recovered only 39% +/- 7%, p < 0.05 vs cardiopulmonary bypass control). Lowering oxygen tension to about 100 mm Hg during reoxygenation avoided conjugated diene production and creatine kinase release, retained normal antioxidant reserve, and improved functional recovery (80% +/- 11%, p < 0.05 vs oxygen tension about 400 mm Hg). These findings show that conventional hyperoxemic cardiopulmonary bypass causes unintended reoxygenation injury in hypoxemic immature hearts that may contribute to myocardial dysfunction after cardiopulmonary bypass and that normoxemic management may be used to surgical advantage. PMID- 7475176 TI - Studies of hypoxemic/reoxygenation injury: with aortic clamping. X. Exogenous antioxidants to avoid nullification of the cardioprotective effects of blood cardioplegia. AB - This study tests the hypothesis that reoxygenation of cyanotic immature hearts when starting cardiopulmonary bypass produces an "unintended" reoxygenation injury that (1) nullifies the cardioprotective effects of blood cardioplegia and (2) is avoidable by adding antioxidants N-(2-mercaptopropionyl)-glycine plus catalase to the cardiopulmonary bypass prime. Twenty immature piglets (2 to 3 weeks) underwent 30 minutes of aortic clamping with a blood cardioplegic solution that was hypocalcemic, alkalotic, hyperosmolar, and enriched with glutamate and aspartate during 1 hour of cardiopulmonary bypass. Of these, six piglets did not undergo hypoxemia (blood cardioplegic control) and 14 others remained hypoxemic (oxygen tension about 25 mm Hg) for up to 2 hours by lowering ventilator fraction of inspired oxygen before reoxygenation on cardiopulmonary bypass. The primary solution of the cardiopulmonary bypass circuit was unchanged in eight piglets (no treatment) and supplemented with the antioxidants N-(2-mercaptopropionyl)-glycine (80 mg/kg) and catalase (5 mg/kg) in six others (N-(2-mercaptopropionyl)-glycine and catalase). Myocardial function (end-systolic elastance), lipid peroxidation (myocardial conjugated diene production), and antioxidant reserve capacity were evaluated. Blood cardioplegic arrest produced no biochemical or functional changes in nonhypoxemic control piglets. Reoxygenation caused an approximate 10 fold increase in conjugated production that persisted throughout cardiopulmonary bypass, lowered antioxidant reserve capacity 86% +/- 12%, and produced profound myocardial dysfunction, because end-systolic elastance recovered only 21% +/- 2%. Supplementation of the cardiopulmonary bypass prime with N-(2-mercaptopropionyl) glycine and catalase reduced lipid peroxidation, restored antioxidant reserve capacity, and allowed near complete functional recovery (80% +/- 8%).** Lipid peroxidation (conjugated diene) production was lower during warm blood cardioplegic reperfusion than during induction in all reoxygenated hearts, which suggests that blood cardioplegia did not injure reoxygenated myocardium. We conclude that reoxygenation of the hypoxemic immature heart causes cardiac functional and antioxidant damage that nullifies the cardioprotective effects of blood cardioplegia that can be avoided by supplementation of the cardiopulmonary bypass prime with antioxidants (*p < 0.05 vs blood cardioplegic control; **p < 0.05 vs reoxygenation). PMID- 7475177 TI - Studies of hypoxemic/reoxygenation injury with aortic clamping: XI. Cardiac advantages of normoxemic versus hyperoxemic management during qardiopulmonary bypass. AB - The conventional way to start cardiopulmonary bypass is to prime the cardiopulmonary bypass circuit with hyperoxemic blood (oxygen tension about 400 mm Hg) and deliver cardioplegic solutions at similar oxygen tension levels. This study tests the hypothesis that an initial normoxemic oxygen tension strategy to decrease the oxygen tension-dependent rate of oxygen free radical production will, in concert with normoxemic blood cardioplegia, limit reoxygenation damage and make subsequent hyperoxemia (oxygen tension about 400 mm Hg) safer. Thirty five immature (3 to 5 kg, 2 to 3 week old) piglets underwent 60 minutes of cardiopulmonary bypass. Eleven control studies at conventional hyperoxemic oxygen tension (about 400 mm Hg) included six piglets that also underwent 30 minutes of blood cardioplegic arrest. Of 25 studies in which piglets were subjected to up to 120 minutes of ventilator hypoxemia (reducing fraction of inspired oxygen to 5% to 7%; oxygen tension about 25 mm Hg), 11 underwent either abrupt (oxygen tension about 400 mm Hg, n = 6) or gradual (increasing oxygen tension from 100 to 400 mm Hg over a 1-hour period, n = 5) reoxygenation without blood cardioplegia. Fourteen others underwent 30 minutes of blood cardioplegic arrest during cardiopulmonary bypass. Of these, nine were reoxygenated at oxygen tension about 400 mm Hg, and five others underwent normoxemic cardiopulmonary bypass and blood cardioplegia (oxygen tension about 100 mm Hg) with systemic oxygen tension raised to 400 mm Hg after aortic unclamping. Measurements of lipid peroxidation (conjugated dienes and antioxidant reserve capacity) and contractile function (pressure-volume loops, conductance catheter, end-systolic elastance) were made before and during hypoxemia and 30 minutes after reoxygenation. Hyperoxemic cardiopulmonary bypass did not produce oxidant damage or reduce functional recovery after cardiopulmonary bypass in nonhypoxemic controls. In contrast, abrupt and gradual reoxygenation without blood cardioplegia produced significant lipid peroxidation (84% increase in conjungated dienes), lowered antioxidant reserve capacity 68% +/- 5%, 44% +/- 8%, respectively, and decreased functional recovery 75% +/- 6% (p < 0.05), 66% +/- 4% (p < 0.05). Similar impairment followed abrupt reoxygenation before blood cardioplegic myocardial management, because conjungated diene production increased 13-fold, antioxidant reserve capacity fell 40%, and contractility recovered only 21% +/- 2% (p < 0.05). Conversely, normoxemic induction of cardiopulmonary bypass and blood cardioplegic myocardial management reduced conjungated diene production 73%, avoided impairment of antioxidant reserve capacity, and resulted in 58% +/- 11% recovery of contractile function.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 7475178 TI - Studies of hypoxemic/reoxygenation injury: with aortic clamping. XII. Delay of cardiac reoxygenation damage in the presence of cyanosis: a new concept of controlled cardiac reoxygenation. AB - Twenty-one immature piglets (< 3 weeks old) underwent 30 minutes of aortic clamping with hypocalcemic glutamate/aspartate blood cardioplegia. Six piglets underwent hyperoxemic cardiopulmonary bypass and blood cardioplegia without preceding hypoxemia (control). Fifteen piglets became hypoxemic (oxygen tension about 25 mm Hg) for up to 2 hours by decreasing ventilator fraction of inspired oxygen to 6% to 7% before cardiopulmonary bypass. Of these, six piglets underwent 5 minutes of abrupt hyperoxemic uncontrolled reoxygenation by starting cardiopulmonary bypass at oxygen tension of about 400 mm Hg before they received oxygen tension of about 400 mm Hg blood cardioplegia. Nine others underwent controlled cardiac reoxygenation by starting cardiopulmonary bypass at ambient oxygen tension (about 25 mm Hg) followed 5 minutes later by 30 minutes of cardiopulmonary bypass at normoxemic oxygen tension (about 100 mm Hg) before raising oxygen tension to about 400 mm Hg. Myocardial function after cardiopulmonary bypass was evaluated from end-systolic elastance by conductance catheter, oxidant damage was estimated by measuring transcoronary conjugated diene levels to detect lipid peroxidation, and antioxidant reserve capacity was determined by measuring malondialdehyde produced from myocardium incubated with the oxidant t-butylhydroperoxide. Hyperoxemic cardiopulmonary bypass and blood cardioplegia preserved myocardial function and produced no oxidant damage in nonhypoxemic piglets. In contrast, uncontrolled reoxygenation at oxygen tension about 400 mm Hg, followed by blood cardioplegia, resulted in marked conjugated dienes production (42 +/- 4* vs 3 +/- 1) A233 nm/min/100 g during blood cardioplegic induction, reduced antioxidant reserve capacity malondialdehyde at 4 mmol/L t-butylhydroperoxide; 1342 +/- 59* vs 958 +/- 50 nmol/g protein) and caused profound myocardial dysfunction; end-systolic elastance recovered only 21% +/- 2%* despite a blood cardioplegic regimen that was cardioprotective in nonhypoxemic piglets. Conversely, controlled cardiac reoxygenation reduced lipid peroxidation (conjugated dienes production was 2 +/- 1**), restored antioxidant reserve capacity (malondialdehyde at 4 mmol/L t-butylhydroperoxide; 982 +/- 88**), and allowed near-complete (83 +/- 8%**) functional recovery. We conclude that reoxygenation of the hypoxemic immature heart by initiating conventional hyperoxemic cardiopulmonary bypass causes oxidant damage characterized by lipid peroxidation, reduced antioxidant reserve capacity, and results in functional depression that nullifies the cardioprotective effects of blood cardioplegia. These changes can be reduced by starting cardiopulmonary bypass at the ambient oxygen tension of the hypoxemic subject and delaying subsequent reoxygenation until blood cardioplegic induction by controlled cardiac reoxygenation (*p < 0.05 vs control; **p < 0.05 vs uncontrol reoxygenation) and analysis of variance. PMID- 7475179 TI - Studies of hypoxemic/reoxygenation injury: with aortic clamping. XIII. Interaction between oxygen tension and cardioplegic composition in limiting nitric oxide production and oxidant damage. AB - This study tests the interaction between oxygen tension and cardioplegic composition on nitric oxide production and oxidant damage during reoxygenation of previously cyanotic hearts. Of 35 Duroc-Yorkshire piglets (2 to 3 weeks, 3 to 5 kg), six underwent 30 minutes of blood cardioplegic arrest with hyperoxemic (oxygen tension about 400 mm Hg), hypocalcemic, alkalotic, glutamate/aspartate blood cardioplegic solution during 1 hour of cardiopulmonary bypass without hypoxemia (control). Twenty-nine others were subjected to up to 120 minutes of ventilator hypoxemia (oxygen tension about 25 mm Hg) before reoxygenation on CPB. To simulate routine clinical management, nine piglets underwent uncontrolled cardiac reoxygenation, whereby cardiopulmonary bypass was started at oxygen tension of about 400 mm Hg followed by the aforementioned blood cardioplegic protocol 5 minutes later. All 20 other piglets underwent controlled cardiac reoxygenation, whereby cardiopulmonary bypass was started at the ambient oxygen tension (about 25 mm Hg), and reoxygenation was delayed until blood cardioplegia was given. The blood cardioplegia solution was kept normoxemic (oxygen tension about 100 mm Hg) in 10 piglets and made hyperoxemic (oxygen tension about 400 mm Hg) in 10 others. The cardioplegic composition was also varied so that the cardioplegic solution in each subgroup contained either KCl only (30 mEq/L) or components that theoretically inhibit nitric oxide synthase by including hypocalcemia, alkalosis, and glutamate/aspartate. Function (end-systolic elastance) and myocardial nitric oxide production, conjugated diene production, and antioxidant reserve capacity were measured. Blood cardioplegic arrest without hypoxemia did not cause myocardial nitric oxide or conjugated diene production, reduce antioxidant reserve capacity, or change left ventricular functional recovery. In contrast, uncontrolled cardiac reoxygenation raised nitric oxide and conjugated diene production 19- and 13-fold, respectively (p < 0.05 vs control), reduced antioxidant reserve capacity 40%, and contractility recovered only 21% of control levels. After controlled cardiac reoxygenation at oxygen tension about 400 mm Hg with cardioplegic solution containing KCl only, nitric oxide and conjugated diene production rose 16- and 12-fold, respectively (p < 0.05 vs control), and contractility recovered only 43% +/- 5%. Normoxemic (oxygen tension of about 100 mm Hg) controlled cardiac reoxygenation with the same solution reduced nitric oxide and conjugated diene production 85% and 71%, and contractile recovery rose to 55% +/- 7% (p < 0.05 vs uncontrolled reoxygenation). In comparison, controlled cardiac reoxygenation with an oxygen tension of about 400 mm Hg hypocalcemic, alkalotic, glutamate/aspartate blood cardioplegic solution reduced nitric oxide and conjugated diene production 85% and 62%, respectively, and contractility recovered 63% +/- 4% (p < 0.05 vs KCl only).(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 7475180 TI - Reflections--projections! PMID- 7475181 TI - Late hemodynamic results after left ventricular patch repair associated with coronary grafting in patients with postinfarction akinetic or dyskinetic aneurysm of the left ventricle. AB - This study reports hemodynamic, electrophysiologic, and clinical results in 171 patients (157 men and 14 women, mean age 57 +/- 8 years) 1 year after endoventricular circular patch repair and coronary grafting for postinfarction left ventricular dyskinetic or akinetic aneurysm. All patients had hemodynamic and electrophysiologic study before the operation and early and 1 year after the operation. The vast majority of aneurysms were anterior (n = 166), with a mean delay from infarction of 43 +/- 50 months. Fifty-two percent of patients were in New York Heart Association class III or IV, and preoperative ejection fraction was less than 40% in the majority of them (75%). Preoperative clinical ventricular tachycardia was present in 25 patients and was inducible in 59 patients. All patients had endoventricular circular patch repair with a synthetic (n = 99) or autologous patch (n = 72); 96% had associated coronary grafting with a mean number of bypass grafts of 1.9 +/- 0.9. Results at 1 year demonstrated a significant increase in ejection fraction (from 36% +/- 13% to 46% +/- 12% (p < 0.0001) and a significant reduction in ventricular volumes (end-diastolic volume index from 116 +/- 5 to 94 +/- 29 ml/m2 and end-systolic volume index from 77 +/- 45 to 53 +/- 25 ml/m2, p < 0.0001). New York Heart Association functional classification was significantly improved (2.6 +/- 0.9 vs 1.4 +/- 0.6, p < 0.0001) and ventricular tachycardias were almost suppressed (no documented clinical ventricular tachycardias and 8% incidence of inducible ventricular tachycardias after 1 year, chi 2 < 0.001). Patients who benefit most from the operation are those with more severe preoperative left ventricular dysfunction (i.e., ejection fraction < 30%), more frequent ventricular arrhythmias, and larger ventricular volumes. At regression analysis, critical disease of the right coronary artery was the only independent predictor of unsatisfactory pump improvement (as evaluated by postoperative increase of ejection fraction < 10 absolute points). In conclusion, in our large series of patients operated on by one surgical team between 1988 and 1993, who were studied hemodynamically both before and after the operation, endoventricular circular patch repair of left ventricular aneurysm associated with coronary grafting definitely improves left ventricular pump function and clinical status 1 year after the operation. PMID- 7475183 TI - Postinfarction ventricular septal rupture: repair by endocardial patch with infarct exclusion. AB - A novel operative technique for postinfarction ventricular septal defect has been used in 44 consecutive patients. The operation consists of excluding rather than excising the infarcted septum and ventricular walls. This is accomplished by performance of a left ventriculotomy through the infarcted muscle and securing a glutaraldehyde-fixed bovine pericardium patch to the endocardium of the left ventricle all around the infarcted myocardium. The ventriculotomy is simply closed over the pericardial patch. There were 21 men and 23 women whose mean age was 69 +/- 7 years. Twenty-nine patients were in cardiogenic shock at the time of operation. All patients had Doppler echocardiography and coronary angiography before operation. All but two patients were operated on during the acute phase of the myocardial infarction. There were six operative deaths. Postoperative complications included renal failure in 10 patients and respiratory failure in 18. Severe right ventricular dysfunction was the only independent predictor of operative mortality. Patients have been followed up for a mean of 40 +/- 34 months. There have been six late deaths and three of these were because of cardiac problems. The actuarial survival at 6 years was 66% +/- 7%. Only one patient had a small residual ventricular septal defect. Late postoperative assessment of ventricular function by echocardiography revealed that most patients had normal or mild impairment of right ventricular function and mild or moderate impairment of left ventricular function. Repair of acute postinfarction ventricular septal defect by endocardial patch with infarct exclusion of the left ventricule probably avoids additional damage to the right ventricle, remodels the acutely infarcted left ventricle, and enhances survival. PMID- 7475182 TI - Improvement of outcomes after coronary artery bypass. A randomized trial comparing intraoperative high versus low mean arterial pressure. AB - BACKGROUND: The objective of this randomized clinical trial of elective coronary artery bypass grafting was to investigate whether intraoperative mean arterial pressure below autoregulatory limits of the coronary and cerebral circulations was a principal determinant of postoperative complications. The trial compared the impact of two strategies of hemodynamic management during cardiopulmonary bypass on outcome. Patients were randomized to a low mean arterial pressure of 50 to 60 mm Hg or a high mean arterial pressure of 80 to 100 mm Hg during cardiopulmonary bypass. METHODS: A total of 248 patients undergoing primary, nonemergency coronary bypass were randomized to either low (n = 124) or high (n = 124) mean arterial pressure during cardiopulmonary bypass. The impact of the mean arterial pressure strategies on the following outcomes was assessed: mortality, cardiac morbidity, neurologic morbidity, cognitive deterioration, and changes in quality of life. All patients were observed prospectively to 6 months after the operation. RESULTS: The overall incidence of combined cardiac and neurologic complications was significantly lower in the high pressure group at 4.8% than in the low pressure group at 12.9% (p = 0.026). For each of the individual outcomes, the trend favored the high pressure group. At 6 months after coronary bypass for the high and low pressure groups, respectively, total mortality rate was 1.6% versus 4.0%, stroke rate 2.4% versus 7.2%, and cardiac complication rate 2.4% versus 4.8%. Cognitive and functional status outcomes did not differ between the groups. CONCLUSION: Higher mean arterial pressures during cardiopulmonary bypass can be achieved in a technically safe manner and effectively improve outcomes after coronary bypass. PMID- 7475184 TI - Reconstruction of the mitral anulus. A ten-year experience. AB - Reconstruction of the mitral anulus was done in 93 patients because of extensive calcification in 24, infective endocarditis with annular abscess in 27, damaged anulus as a result of previous valve replacement in 36, and rupture of the posterior wall of the left ventricle after mitral valve replacement in 6. The reconstruction was done with fresh autologous pericardium in 30 patients, glutaraldehyde-fixed bovine pericardium in 56, and Dacron graft in 7. An appropriately tailored patch was sutured to the endocardium of the left ventricle or to fibrous skeleton of the heart, or to both. Sixty patients had had previous operation; most patients were in New York Heart Association functional class IV and 15 were in shock. The mitral valve was repaired in 10 patients and replaced in 83. Other procedures were aortic valve replacement in 40, tricuspid valve repair in 25, coronary artery bypass in 12, and replacement of the ascending aorta in 2. Eight patients died in the perioperative period. Postoperative complications were common. Patients were followed up from 12 to 96 months, with a mean of 30 months. There have been 18 late deaths, mostly cardiac. The actuarial survival at 5 years was 68% +/- 6%. A total of 8 patients have required reoperation: 4 because of endocarditis, 2 because of bioprosthetic valve failure, and 2 because of patch dehiscence. The freedom from reoperation was 80% +/- 8% at 5 years. The freedom from patch dehiscence was 95% +/- 4% at 5 years. Reconstruction of the mitral anulus has been an extremely useful operative technique for patients with complex mitral valve disease. PMID- 7475185 TI - Atrioventricular valve repair using externally adjustable flexible rings. AB - Initial results obtained with a new flexible ring, adjustable from outside of the heart after interruption of extracorporeal circulation, are presented. Twenty five rings have been inserted in 20 patients, 14 in the mitral position and 11 in the tricuspid position. In 8 of the 14 patients receiving mitral annuloplasty, other standard mitral valve repair techniques were used. Adjustment, assisted by intraoperative transesophageal color Doppler echocardiography, was done for 10 (71%) of the mitral rings and for 8 (73%) of the tricuspid rings. Residual mitral regurgitation disappeared or became minimal in 9 (90%) patients, and a mild regurgitation remained in 1. In all patients who received tricuspid rings regurgitation was abolished after the adjustment. There was no hospital or late mortality. After a maximum follow-up of 6 months results are comparable in the tricuspid and mitral positions and echocardiographic evaluation revealed stable competent valves in all patients but one, who underwent reoperation because of failure of a mitral valve chordae shortening procedure. The use of externally adjustable rings for the mitral and tricuspid valves is a safe alternative for atrioventricular valve annuloplasty and has the additional advantage of reducing postrepair regurgitation. PMID- 7475186 TI - Revascularization of the circumflex artery with the pedicled right internal thoracic artery: clinical functional and angiographic midterm results. AB - Retroaortic crossing of the pedicled right internal thoracic artery for revascularization of the circumflex artery used in combination with a pedicled left internal thoracic artery anastomosed to the left anterior descending artery and its branches is an attractive technique to achieve an extensive arterial revascularization of the left ventricle. However, there is a suspicion that pulling the right internal thoracic artery through the transverse sinus could compromise its blood flow capacity and patency. Between January 1990 and July 1994 this technique was applied in 256 patients (202 men, 54 women; average age 62 years, range 31 to 80 years). Sixty-one patients had two-vessel disease and 195 had three-vessel disease. Seventeen patients were undergoing a reoperation. Twenty-two had a left ventricular ejection fraction of 40% or less. Thirty had diabetes. Twenty-eight had morbid obesity. The right internal thoracic artery was directed to the circumflex artery (259 anastomoses) through the transverse sinus and the left internal thoracic artery was anastomosed to the left anterior descending artery and its branches (375 anastomoses) in all patients. The 195 patients with three-vessel disease received additional coronary artery bypass grafts to the right coronary artery (93 saphenous vein grafts, 89 free inferior epigastric artery grafts, 12 pedicled right gastroepiploic artery grafts). In total, the 256 patients received 833 distal anastomoses (average 3.2, maximum 5 per patient) and 634 distal anastomoses were internal thoracic artery anastomoses (average 2.4, maximum 4 per patient). Three patients died early and eight had a nonfatal myocardial infarction. Seven patients needed postoperative intraaortic balloon pump support. Six patients underwent early reoperation because of excessive bleeding. Sternal dehiscence occurred in four patients. One of these four patients died of the complication 10 months after the operation. No patient was lost to follow-up (average 33 months). During follow-up, two sudden deaths and six noncardiac deaths occurred. Two patients had a nonfatal myocardial infarction and 12 had recurrence of angina. There were no late reoperations. One patient underwent a successful percutaneous balloon angioplasty of a native left anterior descending artery. Seventy-four patients, enrolled in prospective angiographic studies, underwent a postoperative recatheterization (average 13.2 months, range 6 to 58 months). Seventy-three of the 74 right internal thoracic artery grafts were patent. In comparison, 74 of 74 of the left internal thoracic artery grafts (106/107 anastomoses) were patent. Maximal stress thallium-201 scintigraphy results, obtained in 25 of those patients, did not reveal ischemia in the area of the circumflex artery.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 7475187 TI - Is body size the cause for poor outcomes of coronary artery bypass operations in women? AB - Although small body size and coronary artery diameter are recognized as major contributors to the increased risk of coronary artery bypass grafting in women, few studies have established the independent influence of body size and gender on outcome. We studied 7025 consecutive patients (5694 men, 1331 women) undergoing isolated coronary artery bypass grafting between 1990 and 1994. Women were older, had higher preoperative prevalences of urgent operation because of unstable angina, diabetes, peripheral vascular disease, hypertension, and single-vessel coronary artery disease (p < 0.0001), and a lower prevalence of left ventricular ejection fraction 40% or less (p < 0.0001). The prevalences of operative mortality (men, 1.8%; women, 3.5%), low-output syndrome (men, 6.6%; women, 14.8%), and myocardial infarction (men, 2.8%; women, 5.5%) were higher in women (p < 0.0001). Patients were divided into quartiles for body surface area, weight, height, and body mass index. For both men and women, there was no difference in operative mortality between the highest and lowest quartiles of body size. Women, however, had a higher prevalence of operative mortality than men in the lower quartiles of body surface area, height, and weight and in the higher quartiles of body mass index. Among men, the prevalence of low-output syndrome increased (p < 0.0001) with decreasing body surface area, weight, and body mass index, suggesting that body size did influence the prevalence of low-output syndrome. However, women had a higher prevalence of low-output syndrome than men in every category and quartile of body size (p < 0.0001). Multivariable analysis identified gender as a significant determinant of operative mortality (odds ratio 1.83, 95% confidence interval 1.27 to 2.64) and low-output syndrome (odds ratio 2.52, 95% confidence interval 2.05 to 3.11). When multivariable adjustments were made for body size and preoperative risk factors, gender remained a predictor of both operative mortality and low-output syndrome. Multivariable assessment of risk for men and women separately identified that urgent operation was a predictor of operative mortality (odds ratio 2.52, 95% confidence interval 1.32 to 5.61) and low-output syndrome (odds ratio 1.57, 95% confidence interval 1.14 to 2.17) in women but not men. In conclusion, the increased risk of coronary artery bypass grafting in women may be explained in part by dramatic differences in preoperative risk factors between men and women. In both men and women, small body size did not increase the risk of operative mortality, but may have contributed to the risk of low-output syndrome.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 7475188 TI - Results of an autologous tissue reconstruction for replacement of obstructed extracardiac conduits. AB - Between May 1983 and March 1, 1995, 50 patients had replacement of an obstructed pulmonary ventricle-pulmonary artery conduit with an autologous tissue reconstruction in which a prosthetic roof was placed over the fibrous tissue bed of the explanted conduit. The roof was constructed with xenograft pericardium (most recently) (n = 42), homograft dura mater (n = 5), or Dacron fabric (n = 3). Patient ages ranged from 5 to 34 years (median 16 years). The explanted conduits were Hancock conduits (n = 33), Tascon conduits (n = 6), homograft (n = 4), Dacron tube (n = 3), and others (n = 4). Preoperative maximum systolic gradients ranged from 44 to 144 mm Hg (median 78 mm Hg). Thirty-seven concomitant cardiac procedures were done in 29 patients. When a valve was necessary (n = 15), it was possible to place a large-sized valve in the autologous tissue reconstructions (range 22 to 29 mm, median 26 mm). Cardiopulmonary bypass times ranged from 34 to 223 minutes (median 84 minutes), and aortic crossclamp times ranged from 0 (in 32 patients) to 109 minutes (median 0 minutes). Intraoperative postrepair peak systolic gradients from pulmonary ventricle to pulmonary artery ranged from 0 to 33 mm Hg (median 13 mm Hg). There was one early death (2%) in a patient who had additional cardiac procedures. Follow-up was complete in all patients and ranged from 1 month to 11.8 years (median 7.5 years). There were two sudden late deaths: conduits in both were known to be free from obstruction. Forty-four of the 47 surviving patients had evaluation of the gradient by echocardiography or cardiac catheterization 1 month to 11 years (median 7 years) after operation. The gradients ranged from 5 to 45 mm Hg (median 20 mm Hg). None of the conduits developed an obstructive peel, valve obstruction, or valve incompetence. At 10 years, the freedom from reoperation for conduit obstruction was 100%, and freedom from reoperation for any cause was 81%. This technique simplifies conduit replacement, allows for a generous-sized outflow tract, has a low risk, and yields late results that appear superior to those of cryopreserved homografts or other types of extracardiac conduits. PMID- 7475189 TI - The direct effects of 3,5,3'-triiodo-L-thyronine (T3) on myocyte contractile processes. Insights into mechanisms of action. AB - Administration of 3,5,3'-triiodo-L-thyronine (T3) has recently been suggested to acutely improve left ventricular performance. However, the cellular and molecular mechanisms responsible for this improvement in left ventricular function with T3 remained unknown. Accordingly, the present study examined the direct effects of T3 administration on myocyte contractile function and the sarcolemmal systems that might potentially contribute to these effects. In isolated porcine left ventricular myocytes (n = 81), velocity of shortening increased in the presence of 80 pmol/L T3 compared with that in untreated myocytes (117.0 +/- 5.0 versus 77.3 +/- 3.3 microns/sec, p < 0.05). In a separate series of experiments (n = 29), myocyte velocity of shortening increased in the presence of both T3 and beta adrenergic receptor stimulation (25 nmol/L isoproterenol) to greater than that with beta-adrenergic receptor stimulation alone (274.3 +/- 16.9 versus 203.7 +/- 16.2 microns/sec, p < 0.05). Cyclic adenosine monophosphate generation was next examined in isolated myocyte preparations (n = 9). In the presence of T3, no significant increase in cyclic-adenosine monophosphate generation was observed compared with that in untreated myocytes (39.1 +/- 8.3 versus 24.7 +/- 5.8 fmols/myocyte, p = 0.17). However, in the presence of both T3 and beta-adrenergic receptor stimulation, cyclic-adenosine monophosphate generation increased significantly to greater than that with beta-adrenergic receptor stimulation alone (224.4 +/- 61.1 versus 120.1 +/- 35.5 fmoles/myocyte, p < 0.05). Because cyclic-adenosine monophosphate modulates intracellular Ca2+ processes, L-type Ca+2 channel current (patch clamp methods; -picoamp/picofarad, n = 15) and peak intracellular Ca+2 levels (fura 2 ionic measurement, n = 47) were next measured. In the presence of T3, a shift in the activation voltage at peak L-type Ca+2 channel current was observed from baseline (5.5 +/- 1.4 versus 9.0 +/- 1.0 mV, p < 0.05). Furthermore, in the presence of both T3 and beta-adrenergic receptor stimulation, peak L-type Ca+2 channel current (8.9 +/- 0.7 versus 6.3 +/- 1.0 mV, p < 0.05) and peak intracellular Ca+2 levels (189.9 +/- 8.4 versus 171.7 +/- 8.3 nmol/L, p < 0.05) increased compared with values obtained with beta-adrenergic receptor stimulation alone. Important findings from the present study were twofold: (1) T3 improved myocyte contractile processes through a cyclic-adenosine monophosphate-independent mechanism and (2) T3 potentiated the effects of beta adrenergic receptor stimulation transduction by increasing cyclic-adenosine monophosphate production, L-type Ca+2 channel current, and Ca+2 availability to the myocyte contractile apparatus.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 7475190 TI - Influence of vein valves in the development of arteriosclerosis in venoarterial grafts in the rabbit. AB - Coronary saphenous vein grafts in human beings have a more limited long-term patency rate than internal thoracic artery grafts, primarily because of more rapid development of arteriosclerosis. The factors responsible for this increased susceptibility are not completely understood. To test the hypothesis that vein valves may influence this process, we studied 48 hypercholesterolemic rabbits with jugular vein grafts interposed into the carotid arterial circulation. In 24 animals (group A), the vein segments did not contain a vein valve. In the other 24 animals (group B), a vein valve was present. Both groups were further divided in four subgroups of six to be put to death at 2, 4, 6, and 8 weeks after the operation. All animals were fed a 2% cholesterol diet. At postmortem examination, alternate 2 mm sections were either stained with hematoxylin and eosin for histologic and morphometric studies or frozen in liquid nitrogen for immunohistochemistry and in situ hybridization studies. Proliferating cell nuclear antigen immunostaining was used to study cell proliferation. Wall thickness of vein grafts increased with time. During the first 2 weeks intimal and medial thickening was primarily due to an increase in numbers of cells. Between 2 and 6 weeks further intimal and medial thickening occurred, but without additional increase in cell numbers. After 6 weeks, foam cells and lipid deposits started to appear. By 8 weeks, changes identical to those seen in arteriosclerotic plaques in human beings were evident. These changes developed sooner and with more intensity in group B animals (p < 0.01 to 0.001), and they developed faster and with more severity in segments of vein located distal to the valve than in the segments located proximal to the valve (p < 0.001). This is the first controlled experiment demonstrating that the presence of valves in the vein segments is associated with augmented and accelerated intimal changes leading to vein atheromatosis. PMID- 7475192 TI - Recurrence of obliterative bronchiolitis and determinants of outcome in 139 pulmonary retransplant recipients. AB - An international series of pulmonary retransplantation was updated to identify the predictors of outcome and the prevalence and recurrence rate of obliterative bronchiolitis after operation. The study cohort included 139 patients who underwent retransplantation in 34 institutions in North America and Europe between 1985 and 1994. Eighty patients underwent retransplantation because of obliterative bronchiolitis, 34 because of acute graft failure, 13 because of intractable airway complications, 8 because of acute rejection, and 4 because of other indications. Survivors were followed up for a median of 630 days, with 48 patients alive at 1 year, 30 at 2 years, and 16 at 3 years after retransplantation. Actuarial survival was 65% +/- 4% at 1 month, 54% +/- 4% at 3 months, 45% +/- 4% at 1 year, 38% +/- 5% at 2 years, and 36% +/- 5% at 3 years; nonetheless, of 90-day postoperative survivors, 65% +/- 6% were alive 3 years after retransplantation. Life-table and univariate Cox analysis revealed that more recent year of retransplantation (p = 0.009), identical match of ABO blood group (p = 0.01), absence of a donor-recipient cytomegalovirus mismatch (p = 0.04), and being ambulatory immediately before retransplantation (p = 0.04) were associated with survival. By multivariate Cox analysis, being ambulatory before retransplantation was the most significant predictor of survival (p = 0.008), followed by reoperation in Europe (p = 0.044). Complete pulmonary function tests were done yearly in every survivor of retransplantation and bronchiolitis obliterans syndrome stages were assigned. Eleven percent of patients were in stage 3 at 1 year, 20% at 2 years, and 25% at 3 years after retransplantation. Values of forced expiratory volume in 1 second decreased from 1.89 +/- 0.13 L early after retransplantation to 1.80 +/- 0.15 L at 1 year and 1.54 +/- 0.16 L at 2 years (p = 0.006, year 2 versus baseline postoperative value). Most of this decrease occurred in patients who underwent retransplantation because of obliterative bronchiolitis, whereas the pulmonary function of patients who underwent retransplantation because of other conditions did not significantly change. We conclude that survival after pulmonary retransplantation is improving. Optimal results can be obtained in patients who are ambulatory before retransplantation. Compared with recent data after primary lung transplantation, bronchiolitis obliterans syndrome does not appear to recur in an accelerated manner after retransplantation. As long as early mortality as a result of infection can be minimized, pulmonary retransplantation appears to offer a reasonable option in highly selected patients. PMID- 7475191 TI - Myocardial stunning: a therapeutic conundrum. AB - Dobutamine and pyruvate are two inotropic agents with different mechanisms of action. Although both agents alter postischemic myocardial dysfunction, their potential metabolic effects in the setting of in vivo myocardial stunning have not been addressed. In this study, the effects of dobutamine and pyruvate on systolic wall thickening, myocardial phosphorylation potential index, interstitial fluid adenosine level, and myocardial oxygen consumption in in vivo stunned porcine myocardium were assessed. Stunning was induced with a 10-minute occlusion of the left anterior descending coronary artery. After 30 minutes of reperfusion, pigs were treated with either intravenous dobutamine (10 micrograms/kg per minute) or intracoronary pyruvate (1 ml/min, 150 mmol/L solution, pH 7.4). Infusion of both agents resulted in a marked improvement in regional systolic wall thickening. The dobutamine effect, however, produced a marked increase in myocardial oxygen consumption and was associated with an increase in interstitial adenosine caused by myocardial de-energization, because the myocardial phosphorylation potential index ratio decreased from 0.17 +/- 0.02 to 0.09 +/- 0.02 (p < 0.05). In contrast, pyruvate enhanced myocardial energy status, because the myocardial phosphorylation potential index ratio increased from 0.20 +/- 0.03 to 0.55 +/- 0.08 (p < 0.01). These experimental findings suggest that under certain circumstances the use of beta-receptor agonists to treat myocardial stunning may be suboptimal, if not undesirable. Further investigation is warranted to determine the optimum therapy for the stunned heart. PMID- 7475193 TI - Combined lung and liver transplantation in patients with cystic fibrosis. A 4 1/2 year experience. AB - Patients with cystic fibrosis who have end-stage respiratory failure and associated liver cirrhosis have been considered poor candidates for lung transplantation because of high morbidity and mortality resulting from hepatic insufficiency after the operation. Since April 1989, our policy has been to combine heart-lung or lung and liver transplantation in this group of patients. Between June 1990 and March 1995, among 25 patients accepted in the program for combined transplantation, nine died awaiting transplantation and 10 underwent one of the following procedures: heart-lung-liver transplantation (n = 5), en bloc double lung-liver transplantation (n = 1), sequential double lung-liver transplantation (n = 3), and bilateral lobar lung transplantation from a split left lung and reduced liver transplantation (n = 1). There were 5 male and 5 female patients. The ages of the recipients ranged from 10 to 24 years. Mean forced expiratory volume in 1 second was 29% and mean forced vital capacity was 35% of predicted values. All patients were infected with resistant Pseudomonas, three with Pseudomonas cepaceia, and two patients had Aspergillus species in addition. All patients had severe cirrhosis with portal hypertension. Four patients had a history of esophageal variceal bleeding and two had had previous portosystemic shunts. The operation was performed as a two-stage procedure, the intrathoracic operation being completed before the abdominal stage was begun. Cardiopulmonary bypass was used in all patients because of poor clinical condition. Immunosuppression consisted of azathioprine, cyclosporine, and prednisone, as for isolated lung transplantation. There were two perioperative deaths, one caused by primary liver failure and the second by early lung dysfunction. For the first 3 months after transplantation pulmonary infection was the most common cause of morbidity. Other complications included tracheal stenosis (n = 1), bronchial stenosis (n = 1), biliary stricture (n = 2), and severe ascites (n = 3). All were successfully treated. Obliterative bronchiolitis developed in three patients. This was stabilized with FK 506 in two patients; the other patient underwent retransplantation at 38 months but eventually died of bleeding. Actuarial survival was 70% at 1 year and remained unchanged at 3 years. Significant functional improvement was observed in all survivors. For patients who have chronic respiratory failure with advanced cirrhosis, lung transplantation combined with liver transplantation can be performed with a satisfactory outcome. PMID- 7475194 TI - Improved airway healing after lung transplantation. An analysis of 348 bronchial anastomoses. AB - We evaluated various clinical factors to identify predictors of airway complication after lung transplantation. Two hundred twenty-nine consecutive single (n = 110) and bilateral (n = 119) lung transplants were done between September 1988 and August 1994. These 348 bronchial anastomoses were retrospectively analyzed. Airway complication that necessitated clinical intervention affected 33 anastomoses (9.5%) in 29 patients (12.8%). Satisfactory healing was achieved in 22 of these patients by conservative therapy such as one or a combination of dilation, stent, and laser. There were five deaths (2.2%) attributable to airway complications. One patient had an early postoperative death unrelated to airway complication and one patient has a recalcitrant bronchus intermedius stricture. Complication occurred more often in single-lung than in bilateral lung transplants (16/110, 14.4%, versus 17/238, 7.1%; p < 0.05). The use of a mattress suture (21/153, 13.7%) was associated with more frequent complications than was simple interrupted suture (8/122, 6.6%) or figure of-eight suture (4/73, 5.5%) (p < 0.05). For patients in whom airway complications subsequently developed, the duration of postoperative mechanical ventilation was greater than that for those in whom an airway complication did not develop. The prevalence of airway complications as our program evolved was evaluated by separating the 229 transplants into three groups: phase I, the first 77 transplants; phase II, the next 76 transplants; and phase III, the most recent 76 transplants. The airway complication rate per anastomosis was significantly lower in phase III (5/126, 4.0%) than in phase I (12/110, 10.9%; p < 0.05) and phase II (16/112, 14.3%; p < 0.01). The majority of airway complications are successfully treated and rarely fatal. The recent reduction in prevalence of airway complications is likely a result of better maintenance immunosuppression and rejection surveillance. PMID- 7475196 TI - Myoblast transplantation in the porcine model: a potential technique for myocardial repair. AB - The use of transgenic cells transplanted in syngeneic rodents has shown modest success, but allogeneic and xenogeneic transplants have not been uniformly successful. To assess the feasibility of xenogeneic and allogeneic myoblast transplantation, we subjected seven adult swine to transplantation of murine atrial tumor cells (xenogeneic), neonatal porcine myocytes (allogeneic), and human fetal cardiomyocytes into the left ventricular wall. After general anesthesia, isolated cells were injected along the anterior and posterior walls of the porcine left ventricle. All the animals were immuno-suppressed and observed for 1 month after injection, at which time they were killed and analyzed. This report will present results primarily concerned with the success of human cell transfers. In all injected sites examined, the transplanted cells thrived within the host myocardium with no significant rejection. Transplant cells formed close associations with host myocytes that resembled nascent intercalated disks on electron microscopy. These cells also contained myofibrils and other cell architecture resembling the transplanted cell lines. Additionally, these cells appeared to produce an angiogenic influence resulting in the proliferation of the surrounding microvasculature. We believe that these findings indicate successful xenogeneic and allogeneic myoblast cell transplantation in a large animal model. These experiments set the stage for future studies to assess the ability of these cells to form a syncytium, contract, and potentially repair failed myocardium. PMID- 7475195 TI - Inhaled nitric oxide fails to confer the pulmonary protection provided by distal stimulation of the nitric oxide pathway at the level of cyclic guanosine monophosphate. AB - It has been suggested that inhaled nitric oxide gas may be beneficial after lung transplantation, because endogenous levels of pulmonary nitric oxide decline rapidly after reperfusion. However theoretical concerns remain about the formation of highly toxic oxidants during the quenching of nitric oxide by superoxide. To determine whether distal stimulation of the nitric oxide-cyclic guanosine monophosphate pathway at the level of cyclic guanosine monophosphate might confer the beneficial vascular effects of nitric oxide without its potential toxicities, we studied an orthotopic rat left lung transplant model. In this model, hemodynamic and survival measurements can be obtained independent of the native right lung. Lungs were preserved for 6 hours at 4 degrees C in Euro Collins solution alone (control, n = 6) or supplemented with the cyclic guanosine monophosphate analog, 8-(4-chlorophenylthio)-guanosine-3',5'-cyclic guanosine monophosphate (cGMP, n = 4). In additional experiments in which lungs were preserved with Euro-Collins solution alone, inhaled nitric oxide was administered during reperfusion (NO, n = 12). Thirty minutes after transplantation and ligation of the native right pulmonary artery, pulmonary vascular resistance, arterial oxygenation, graft neutrophil infiltration (myeloperoxidase activity), and recipient survival were evaluated. Cyclic guanosine monophosphate decreased pulmonary vascular resistance (1.1 +/- 0.2 vs 12.1 +/- 6.3 mm Hg/ml/min, p < 0.05), improved oxygen tension (369 +/- 56 vs 82.8 +/- 48 mm Hg, p < 0.05), reduced myeloperoxidase activity (1.7 +/- 0.3 vs 3.1 +/- 0.9 delta Abs 460 nm/min, p < 0.05), and improved recipient survival (100% vs 0%, p < 0.005) compared with Euro-Collins solution alone (control group). Animals receiving inhaled nitric oxide during reperfusion did not differ from control animals with respect to any of these parameters. These data suggest that distal stimulation of the nitric oxide-cyclic guanosine monophosphate pathway at the level of cyclic guanosine monophosphate has a protective effect that is not seen with inhaled nitric oxide in the immediate pulmonary reperfusion period. PMID- 7475197 TI - Inhibition of inducible nitric oxide synthase ameliorates rat lung allograft rejection. AB - Recently, the inducible isoform of nitric oxide synthase has been shown to be an important immunomodulation molecule in allograft rejection. We have observed the production of nitric oxide during rejection and the effect of nitric oxide synthase inhibition on allograft rejection in a rat lung transplant model. Rat left lung allotransplants were performed in two strain combinations: brown Norway to-F344 (major histocompatibility complex incompatible); and Lewis-to-F344 (minor loci incompatible) as severe and mild rejection models respectively. Syngeneic F344-to-F344 transplants were performed as a negative control. Nitric oxide production during rejection was determined by measuring the recipient's serum nitrite/nitrate levels as a stable end product of nitric oxide. The progression of rejection was evaluated radiographically and the grade of rejection was determined histologically. After operation, recipients of allotransplantation were randomly divided into two groups and received either aminoguanidine (200 mg/kg, intraperitoneal every 6 hours), a potent inducible nitric oxide synthase inhibitor, or normal saline treatment. The levels of serum nitrite and nitrate in recipients increased in the early phase of rejection in both allotransplant combinations. However, in the terminal phase of rejection, the serum nitrite/nitrate level decreased significantly compared with the peak level in the brown Norway-to-F344 recipients. The serum nitrite/nitrate levels in the syngeneic transplant recipients were normal during the entire observation period. In aminoguanidine-treated animals, serum nitrite/nitrate levels remained normal in both allograft combinations. Significant suppression of rejection in aminoguanidine-treated recipients was observed histologically and radiographically in comparison with untreated recipients in the brown Norway-to F344 combinations. In the Lewis-to-F344 combination, aminoguanidine treatment significantly ameliorated histologic rejection but did not affect radiologic appearance. We therefore conclude nitric oxide is produced during early allograft rejection and may prove to be a marker and mediator of early rejection. The inhibition of inducible nitric oxide synthase results in significant reduction in rat lung allograft rejection. PMID- 7475199 TI - The effect of retrograde cerebral perfusion after particulate embolization to the brain. AB - Neurologic injury as a consequence of cerebral embolism of either air or atherosclerotic debris during cardiac or aortic surgery is still a major cause of postoperative morbidity and mortality. While exploring various means of improving cerebral protection during complex cardiothoracic procedures, we have developed a chronic porcine model to study retrograde cerebral perfusion. We have previously demonstrated that retrograde perfusion results in a small amount of nutritive flow and provides cerebral protection that appears to be superior to simple prolonged hypothermic circulatory arrest. The current study was designed to evaluate the efficacy of retrograde cerebral perfusion in mitigating the effects of particulate cerebral embolism occurring during cardiac surgery. Four groups of pigs (19 to 28 kg) underwent cardiopulmonary bypass with deep hypothermia at an esophageal temperature of 20 degrees C: an antegrade control group (AC, n = 5), an antegrade embolism group (AE, n = 10), a retrograde control group (RC, n = 5), and a retrograde embolism group (RE, n = 10). In addition, because of extreme heterogeneity in outcome in the initial RE group, an additional group of 10 animals underwent embolism and retrograde perfusion at a later time. Embolization was accomplished by injection of 200 mg of polystyrene microspheres (250 to 750 micrograms in diameter) via the aortic cannula into an isolated aortic arch preparation in the AE and RE groups; the control groups received injections of 10 ml of saline solution. After infusion of the microspheres or saline solution, conventional perfusion, with the aortic arch pressure maintained at 50 mm Hg, was continued for a total of 30 minutes in the antegrade groups; in the retrograde groups, retrograde flow was initiated via a cannula positioned in the superior vena cava, and was continued for 25 minutes. Superior vena caval flow was regulated to maintain a sagittal sinus pressure of approximately 30 mm Hg in the retrograde groups, and blood returning to the isolated aortic arch was collected and measured. All animals were allowed to recover and were evaluated daily according to a quantitative behavioral score in which 9 indicates apparently complete normalcy, with lower numbers indicating various degrees of cerebral injury. At the time of planned death on day 6, half of the brain was used for recovery of embolized microspheres after digestion with 10N sodium hydroxide. The other half was submitted for histologic study. Neurologic recovery in both the antegrade and retrograde control groups appeared to be complete, although mild evidence of histologic damage was present in some animals in the retrograde control group.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 7475198 TI - Comparison of perioperative myocardial protection with nifedipine versus nifedipine and metoprolol in patients undergoing elective coronary artery bypass grafting. AB - A randomized study was performed on 70 patients undergoing elective coronary bypass grafting to examine whether the combined infusion of the calcium channel blocker nifedipine (10 micrograms/kg per hour) and the beta 1-blocker metopropol (12 micrograms/kg per hour, n = 34) reduces the prevalence of perioperative myocardial ischemia and arrhythmias. The control group received nifedipine alone (n = 36). In both groups the infusion was started from the onset of extracorporal circulation and maintained over a period of 24 hours. Repeated 12-lead electrocardiographic and 3-channel Holter monitor recordings for 48 hours were used to define perioperative myocardial ischemia (transient ischemic event, myocardial infarction) and arrhythmias (sinus tachycardia, supraventricular tachycardia, atrial flutter/fibrillation, ventricular tachycardia). Hemodynamic parameters were repeatedly assessed for 24 hours and serum enzyme levels (creatine kinase, MB isoenzyme of creatine kinase) for up to 36 hours after the operation. The two groups did not differ significantly with respect to preoperative anamnestic and surgical data. No signs of perioperative myocardial infarction were detected in either group. However, a significantly lower incidence of transient ischemic episodes was observed in the nifedipine metoprolol group than in the nifedipine group (3% vs 11%; p < 0.05). In addition, there was a tendency toward lower creatine kinase MB levels and peak values of creatine kinase and creatine kinase MB in the nifedipine-metoprolol group. With regard to perioperative arrhythmias, there was a significantly lower incidence of sinus tachycardia and atrial flutter/fibrillation in the nifedipine-metoprolol group (9% and 6%) than in the nifedipine group (33% and 27%, p < 0.05). In addition, postoperative heart rate was lower in the nifedipine-metoprolol group starting from the sixth hour after release of the aortic crossclamp (p < 0.05 and p < 0.01, respectively). No other hemodynamic parameters showed significant differences between the two groups and all returned to preoperative levels within 24 hours. In conclusion, perioperative application of nifedipine and metoprolol in patients undergoing elective coronary bypass grafting reduces the prevalence of perioperative myocardial ischemia and arrhythmias without significant negative inotropic effects. The combined infusion of the two drugs appears superior to nifedipine alone in preventing perioperative myocardial ischemia and reducing reperfusion-induced arrhythmias. PMID- 7475200 TI - The role of N omega-nitro-L-arginine in modulation of pulmonary vascular tone in the maturing newborn pig. AB - Current therapeutic modalities for treatment of newborn pulmonary hypertensive crisis include but are not limited to the administration of nitric oxide (endothelium-derived relaxing factor). However, few data are available on the role of endogenously produced endothelium-derived relaxing factor in the modulation of pulmonary vascular tone in the neonate. In the current study, we investigated the acute effects of N omega-nitro-L-arginine (a potent competitive inhibitor of endothelium-derived relaxing factor synthase) on the pulmonary vasculature of anesthesized open-chest 48-hour-old (n = 8) and 2-week-old (n = 7) Yorkshire pigs. After baseline data were acquired, all animals received a 10 mg/kg per minute infusion of N omega-nitro-L-arginine for 10 minutes. To discern distal and proximal pulmonary arterial vessel changes, input mean and characteristic impedance were respectively determined. Pulmonary vascular resistance was also calculated (units determined in dyne.sec.cm-5 plus or minus the standard error of the mean). Results showed N omega-nitro-L-arginine infusion did not significantly alter baseline pulmonary arterial pressure (22,370 +/- 1473 dyne/cm2), pulmonary vascular resistance (5171 +/- 805 dyne.sec.cm-5), input impedance (6343 +/- 806 dyne.sec.cm-5), or characteristic impedance (2073 +/- 418 dyne.sec.cm-5) in 48-hour-old pigs. In 2-week-old pigs, infusion of N omega-nitro L-arginine elevated pulmonary arterial pressure (18,162 +/- 1415 dyne/cm2 versus 23,838 +/- 1810 dyne/cm2, p = 0.015), pulmonary vascular resistance (810 +/- 137 dyne.sec.cm-5 versus 1519 dyne.sec.cm-5, p = 0.030), and input impedance (2302 +/ 251 dyne.sec.cm-5 versus 2900 +/- 255 dyne.sec.cm-5, p = 0.018). Characteristic impedance was not altered in 2-week-old pigs. These data indicate that N omega nitro-L-arginine infusion resulted in pulmonary arteriolar vasoconstriction in 2 week-old pigs, but not in 48-hour-old pigs. This finding suggests that endothelium-derived relaxing factor does not modulate basal pulmonary arteriolar tone during the early newborn period, but does play a significant role in 2-week old pigs. These data also suggest that the functional role for endothelium derived relaxing factor is confined to the distal arteriolar pulmonary bed and does not extend to the larger proximal arterial vessels. PMID- 7475201 TI - Catastrophic complications of the cervical esophagogastric anastomosis. AB - Recent enthusiasm for the cervical esophagogastric anastomosis has arisen because of its perceived low morbidity. Although catastrophic complications of a cervical esophagogastric anastomosis are unusual, they can and do occur, and prevention is possible if the potential for them is recognized. Among 856 patients undergoing a cervical esophagogastric anastomosis after transhiatal esophagectomy, catastrophic cervical infectious complications occurred in 11 patients (1.3%): vertebral body osteomyelitis (1), epidural abscess with neurologic impairment (2), pulmonary microabscesses from internal jugular vein abscess (1), tracheoesophagogastric anastomotic fistula (1), and major dehiscence necessitating anastomotic takedown (6). These complications became manifest from 5 to 85 days after the esophageal resection and reconstruction (mean 19 days). Leakage from a gastric suspension stitch placed in the anterior spinal ligament over the vertebral bodies resulted in a posterior gastric leak and either osteomyelitis or an epidural abscess in three patients, none of whom had evidence of extravasation on the routine barium swallow 10 days after operation. Cervical exploration for a presumed anastomotic leak led to the unexpected discovery of an abscess formed by the stomach and the adjacent wall of the internal jugular vein, which was ligated and resected. One patient without symptoms who was discharged from the hospital with a contained anastomotic leak on the postoperative barium swallow was readmitted 7 days later with a cervical tracheoesophagogastric anastomotic fistula of which he ultimately died. In 6 patients (7% of those who had anastomotic leaks) there was sufficient gastric ischemia or necrosis, or both, to necessitate takedown of the anastomosis and intrathoracic stomach, cervical esophagostomy, and insertion of a feeding tube. As a result of this experience, it is recommended that cervical gastric suspension sutures either be omitted entirely or placed in the fascia over the longus colli muscles anterior to the spine, but not directly into the prevertebral fascia overlying the vertebral bodies or cervical disks. All but minute cervical anastomotic leaks, even if apparently contained, are best drained rather than treated expectantly. Patients who remain febrile and ill after bedside drainage of a cervical esophagogastric anastomosis leak should undergo cervical reexploration in the operating room; major gastric ischemia or necrosis, or both, may warrant takedown of the anastomosis and intrathoracic stomach. PMID- 7475202 TI - Intracardiac thrombus after the Fontan procedure. PMID- 7475203 TI - Repair of long-segment tracheal stenosis in infancy. AB - Long-segment stenosis of the trachea in infancy is a considerable surgical challenge because the infants are generally extremely ill and the airway is small. The optimal type of repair is not clearly defined. This report summarizes our experience with rib cartilage tracheoplasty done with cardiopulmonary bypass. Six patients underwent repair of long-segment tracheal stenosis between September 1987 and September 1994. The mean age was 14 weeks (range 1 to 58 weeks). Patients had stenosis of at least 70% of the tracheal length, typically with complete cartilaginous rings. In all patients stenosis was repaired by placement of a section of rib cartilage as an augmentation patch into the anterior surface of the trachea, which had been incised through the entire length of the stenosis. To avoid distal airway intubation, we used cardiopulmonary bypass for all procedures, with a mean bypass duration of 110 minutes (range 54 to 175 minutes). Mechanical ventilation was required for a median of 11 days after the operation (range 7 to 81 days), and the median postoperative hospital stay was 17 days (range 12 to 180 days). All patients are long-term survivors. Complications included the need for extracorporeal membrane oxygenation to treat ventricular dysfunction in one patient and graft dehiscence requiring revision of the distal graft in another. The latter patient has required several treatments with a bronchoscope for removal of granulation tissue. All other patients are free of symptoms and have normal growth with a mean follow-up of 4.7 years (range 5 months to 7.6 years). We conclude that rib cartilage tracheoplasty for long segment tracheal stenosis provides excellent results in short and intermediate follow-up. In addition, the use of cardiopulmonary bypass allows an unobstructed view of the tiny infant airway and thus permits a precise repair. PMID- 7475204 TI - Experience with repair of congenital heart defects using adjunctive endovascular devices. AB - The use of endovascular devices as an adjunct to repair of congenital heart anomalies represents a novel but unproven therapeutic approach. Intraoperative implantation of pulmonary arterial stents (5 to 15 mm diameter) was done in 11 patients with pulmonary atresia with ventricular septal defect (n = 4), classic tetralogy of Fallot (n = 2), truncus arteriosus (n = 1), hypoplastic left heart syndrome (stage II [n = 1] and stage III [n = 1] Norwood procedure), and miscellaneous pulmonary arterial stenoses (n = 3), as well as in patients with congenital (n = 1) and postoperative (n = 3) pulmonary venous obstruction and in 1 patient with combined pulmonary arterial and venous obstruction. The stents were effective at achieving immediate patency in all patients. There were two early deaths, one related to acute thrombosis of a small-diameter left pulmonary artery stent. Reintervention because of stent-related pulmonary arterial stenosis was frequently necessary. In five of seven patients who survived more than 1 month after implantation of stent size 8 mm or smaller severe stent-related pulmonary arterial obstruction developed. In four of the five patients with pulmonary vein stent implantation intractable obstruction developed, resulting in death in all three patients who had bilateral pulmonary vein stent implantation. Intraoperative occlusion of apical muscular ventricular septal defect with use of a clamshell device inserted from the right atrial approach was accomplished in four patients. One patient who underwent associated aortic arch reconstruction died as a result of left ventricular hypoplasia. The results in the remaining three patients were favorable on the basis of absence of significant late residual intraventricular shunting, left ventricular dysfunction, or arrhythmia. We conclude that recurrent intraluminal obstruction as a result of neointimal hyperplasia appears to be an eventual certainty in currently designed small diameter endovascular stents. For this reason, we would recommend standard surgical techniques for repair of obstructive lesions of the pulmonary arterial confluence to maximize growth potential. Device occlusion of muscular ventricular septal defects is feasible but probably only indicated for complex cases of ventricular septal deficiency that otherwise necessitate a left ventriculotomy. PMID- 7475205 TI - The management of tetralogy of Fallot with pulmonary atresia and diminutive pulmonary arteries. AB - Since September 1991, 14 consecutive patients with tetralogy of Fallot, pulmonary atresia, and diminutive pulmonary arteries have undergone staged repair. All patients had multiple aortopulmonary collateral arteries and the ductus arteriosus was absent in 11. Mean sizes of the right and left pulmonary arteries were 2.2 +/- 0.7 mm and 1.9 +/- 0.8 mm, respectively (range 0.5 to 3.0 mm). Eight patients (57%) have subsequently received complete repair. Age at initial procedure (shunt, right ventricle-pulmonary artery conduit, or direct aorta pulmonary artery anastomosis) in this group was 5.3 +/- 6.8 months. The number of operative procedures to achieve complete repair was 2.9 +/- 0.8 per patient (range 2 to 4). Intraoperative postrepair peak right ventricle-left ventricle pressure ratio was 0.57 +/- 0.17. Six of 8 patients (75%) required additional interventional procedures (mean 1.5 +/- 1.2 per patient) for angioplasty of peripheral pulmonary artery stenoses, coil embolization of aortopulmonary collateral arteries, or intra-operative insertion of intravascular pulmonary artery stents. Mean follow-up from complete repair was 8.7 +/- 8.3 months (range 0.5 to 23.8 months) and is complete. There was one in-hospital death at 45 days, and one late cardiac death at 20.3 months. Six patients had initial palliative operations (unifocalization, right ventricle-pulmonary artery conduit, direct aorta-pulmonary artery anastomosis, or transannular outflow patch) but have not undergone complete repair. Age at initial procedure in this group was 27.9 +/- 56.9 months (range 0.27 to 155 months), and mean follow-up from initial procedure was 10.9 +/- 11.2 months (range 0 to 31.4 months). The operative mortality rate was 33% (2 of 6 patients). There was one late noncardiac death at 5.3 months. Three patients are awaiting further intervention or repair. This experience suggests that complete repair is feasible even in patients with extremely diminutive pulmonary arteries (< or = 3.0 mm). Pulmonary artery growth is facilitated by early (3 to 6 month) establishment of central pulmonary artery flow by right ventricle-pulmonary artery conduit (pulmonary arteries > 1.5 mm) or by direct ascending aorta-pulmonary artery anastomosis (pulmonary arteries < 1.5 mm). Subsequent interventional catheterization and operative procedures as required for pulmonary artery stenoses and coil embolization of collateral arteries allow continued recruitment of central pulmonary arteries and may obviate or minimize the need for unifocalization procedures. PMID- 7475206 TI - Subaortic stenosis in the spectrum of atrioventricular septal defects. Solutions may be complex and palliative. AB - From July 1982 through September 1994, 19 children had operative treatment of subaortic stenosis associated with an atrioventricular septal defect. Specific diagnosis were septum primum defects in 7, Rastelli type A defects in 6, transitional defects in 4, inlet ventricular septal defect with malattached chordae in 1, and tetralogy of Fallot with Rastelli type C defect in 1. Twenty seven operations for subaortic stenosis were performed. Surgical treatment of the outlet lesion was performed at initial atrioventricular septal defect repair in 3 children and in the remaining 16 from 1.2 to 13.1 years (mean 4.9 years, median 3.9 years) after repair. Eighteen of the 19 children had fibrous resection and myectomy for relief of obstruction. Seven children had an associated left atrioventricular valve procedure. One child received an apicoaortic conduit. Seven children (36.8%) required 8 reoperations for previously treated subaortic stenosis. Time to the second procedure was 2.8 to 7.4 years (mean 4.9 years). Follow-up is 0.4 to 14.0 years (median 5.6 years). Six-year actuarial freedom from reoperation is 66% +/- 15%. The angle between the plane of the outlet septum and the plane of the septal crest was measured in 10 normal hearts (86.4 +/- 13.7) and 10 hearts with atrioventricular septal defects (22.2 +/- 26.0; p < 0.01). The outflow tract can be effectively shortened, widened, and the angle increased toward normal by augmenting the left side of the superior bridging leaflet and performing a fibromyectomy. CONCLUSION: Standard fibromyectomy for subaortic stenosis in children with atrioventricular septal defects leads to a high rate of reoperation. Leaflet augmentation and fibromyectomy may decrease the likelihood of reoperation. PMID- 7475208 TI - Results of Norwood's operation for lesions other than hypoplastic left heart syndrome. AB - Norwood's operation provides satisfactory palliation for neonates with hypoplastic left heart syndrome. The dominant physiologic features of hypoplastic left heart syndrome, ductal dependency of the systemic circulation and parallel pulmonary and systemic circulations, are shared by a multitude of other less common congenital heart malformations. Theoretically, these should be equally amenable to palliation by Norwood's operation. Between January 1990 and June 1994, 60 neonates with malformations other than hypoplastic left heart syndrome underwent initial surgical palliation by Norwood's procedure. Diagnoses included single left ventricle with levo-transposition of the great arteries (12); critical aortic stenosis (8); complex double-outlet right ventricle (8); interrupted aortic arch with ventricular septal defect and subaortic stenosis (7); ventricular septal defect, subaortic stenosis, and coarctation of the aorta (7); aortic atresia with large ventricular septal defect (6); tricuspid atresia with transposition of the great arteries (6); heterotaxy syndrome with subaortic obstruction (3); and other (3). There were 10 hospital deaths and 50 survivors (83% survival). After the introduction of inspired carbon dioxide therapy into the postoperative management protocol (1991), 42 of 47 patients survived (89% survival). Mortality was independent of diagnosis and essentially the same as that for hypoplastic left heart syndrome. With minor technical modifications, Norwood's operation provides satisfactory initial palliation for a wide variety of malformations characterized by ductal dependency of the systemic circulation in anticipation of either a Fontan procedure or a biventricular repair. PMID- 7475207 TI - Surgical management of complete atrioventricular septal defects. A twenty-year experience. AB - Creation of a competent left atrioventricular valve is a cornerstone in surgical repair of complete atrioventricular septal defects. To identify risk factors for mortality and failure of left atrioventricular valve repair and to determine the impact of cleft closure on postoperative atrioventricular valve function, we retrospectively analyzed hospital records of 203 patients between January 1974 and January 1995. Overall early mortality was 7.9%. Operative mortality decreased significantly over the period of the study from 19% (4/21) before 1980 to 3% (2/67) after 1990 (p = 0.03). Ten-year survival including operative mortality was 91.3% +/- 0.004% (95% confidence limit): all survivors are in New York Heart Association class I or II. Preoperative atrioventricular valve regurgitation was assessed in 203 patients by angiography or echocardiography and was trivial or mild in 103 (52%), moderate in 82 (41%), and severe in 18 (8%). Left atrioventricular valve cleft was closed in 93% (189/203) but left alone when valve leaflet tissue was inadequate and closure of the cleft might cause significant stenosis. Reoperation for severe postoperative left atrioventricular valve regurgitation was necessary in eight patients, five of whom initially did not have closure of the cleft and three of whom had cleft closure. Six patients had reoperation with annuloplasty and two patients required left atrioventricular valve replacement. Five patients survived reoperation and are currently in New York Heart Association class I or II. On most recent evaluation assessed by angiography or echocardiography (a mean of 59 months after repair), left atrioventricular valve regurgitation was trivial or mild in 137 of the 146 survivors (94%) examined; none had moderate or severe left atrioventricular valve stenosis. By multiple logistic regression analysis, strong risk factors for early death and need for reoperation included postoperative pulmonary hypertensive crisis, immediate postoperative severe left atrioventricular valve regurgitation, and double-orifice left atrioventricular valve. These results indicate that complete atrioventricular septal defects can be repaired with low mortality and good intermediate to long-term results. Routine approximation of the cleft is safe and has a low incidence of reoperation for left atrioventricular valve regurgitation. PMID- 7475209 TI - Thrombosis of the pulmonary artery stump after distal ligation. PMID- 7475211 TI - Runoff dependence of inferior epigastric artery grafts in coronary artery operations. PMID- 7475210 TI - Fatal pulmonary hypertension after heart transplantation: sequelae of postactinic pulmonary vascular changes. PMID- 7475212 TI - Vascular reactivity of the gastroepiploic artery graft. PMID- 7475213 TI - Ostioplasty for isolated coronary artery ostial stenosis. PMID- 7475214 TI - Methodologic issues in studying glucocorticoid-induced damage to neurons. PMID- 7475215 TI - Fractal analysis of the electromyographic interference pattern. AB - Evaluation of motor unit recruitment is an important component of the clinical EMG exam. Typically this is assessed qualitatively using auditory features and estimates of the visual complexity of the EMG waveform. Recent advances in nonlinear dynamics have led to the development of the concept of fractals which can be used to quantify complexity and space filling features of various structures. This study was undertaken to determine if the normal EMG interference pattern (IP) has fractal characteristics that might be helpful in quantitative analysis. EMG activity was recorded from the 9 normal biceps muscles as force was varied from 10 to 90% of maximal. Using a box count algorithm, the fractal dimension was calculated. The EMG IP displays fractal characteristics with a dimension that is highly correlated with force and ranges from 1.1 to 1.4 as force increases from 10 to 90% MVC. The fractal dimension (FD)-force relationship is similar to that observed with other methods of IP analysis and suggests that the fractal dimension can be used to quantify and capture the essence of the 'complexity' of motor unit recruitment patterns. PMID- 7475216 TI - Stereoscopic and biplanar microphotography of Golgi-impregnated neurons: a correlative study using conventional and real-time, direct-image confocal microscopies. AB - A correlative study of neuronal reconstruction methods was made using both conventional (non-confocal) and real-time confocal microscopies. Simple and sophisticated (totally automated) methods are described for making both biplanar microphotographs using conventional transmitted light, and stereoscopic microphotographs using real-time confocal microscopy of Golgi-impregnated neurons. Confocal microscopy discriminates against out-of-focus layers to produce optical sections which can be summed on photographic film to obtain neuronal reconstructions. Biplanar images are obtained by fusion, using a stereo-viewer, of two adjacent optical sections obtained with a conventional transmitted light microscope. Stereoscopic and biplanar microphotographs of 3-day-old chick neurons are presented. PMID- 7475217 TI - Analysis of differences between conditioned and control reflex series in EMG recordings. AB - Two methods are presented for the data analysis of signals derived by subtracting conditioned from control reflex data in full-wave rectified and averaged electromyographic (EMG) signals. The first method uses the ratios of the mean amplitudes and standard errors of the mean (S.E.M.s) (i.e. Student's t values) of a series of data points in such a difference signal, and deals with the detection of latencies of reflex components which are susceptible to a conditioning stimulus or experimental situation. The second method applies a modified cumulative sum (CUSUM) technique to full-wave rectified difference signals. This modified CUSUM technique determines the magnitude of the effects of the conditioning situation above that expected due to chance fluctuations, taking into account the effect of reflex modulations on such chance fluctuations in a post-stimulus period. The modified CUSUM technique proved particularly useful when various subtle but consistent, opposing changes occurred sequentially in the conditioned series thus yielding a complicated pattern of effects in a difference signal, with a low signal-to-noise ratio. PMID- 7475218 TI - A method for detecting the time course of correlation between single-unit activity and EMG during a behavioral task. AB - The chance that a change in excitability of one neuron leads to a change in excitability of another is likely to vary within a single volitional act. This temporal variability in functional connectivity is impossible to assess with standard analytical procedures to accurately that measure the correlation between such elements. This reports describes a technique designed to overcome this limitation by expressing a correlation measure calculated repeatedly in short epochs throughout a behavioral trial. The activity of two elements, a motor cortical neuron and a shoulder muscle, that might take place during a drawing task was first simulated so that the correlation could be manipulated. Various correlation algorithms (standard cross-correlation, spike-triggered average, impulse-response function, impulse-response surface) were tested with these data. Spike trains from a monkey's motor cortex and rectified EMG from its posterior deltoid muscle were compared using the same techniques and shown to have a correlation that changed in a characteristic manner throughout a task that required the monkey to draw a sinusoid. PMID- 7475219 TI - Local cerebral glucose utilization in perfusion-fixed rat brains. AB - Local cerebral glucose utilization (LCGU) was measured in 75 cortical areas and nuclei of adult, 3-4-month-old Wistar rats, using the [14C]2-deoxyglucose (2-DG) technique. Measurement of total brain radioactivity content was not significantly different in unfixed material compared to fixed brain tissue. Values of LCGU derived from fresh, unfixed material were compared with values obtained from rats fixed by perfusion 45 min after the [14C]2-DG bolus injection with phosphate buffered 3.3% paraformaldehyde at room temperature. In the fixed material, the mean LCGU of all brain regions was significantly increased by about 25% compared with the unfixed specimen due to tissue shrinkage of 7.2% in the fixed brains. Shrinkage leading to a higher volume density of [14C]2-deoxyglucose-6-phosphate in brain tissue results in a higher grain density in the respective autoradiographs. The wash-out of blood-borne [14C]2-DG is negligible except for blood-rich structures like the pineal gland and the choroid plexus. PMID- 7475220 TI - The grasping test: a simple behavioral method for objective quantitative assessment of peripheral nerve regeneration in the rat. AB - Behavioral recovery is the major goal of neuroscience research on the peripheral nervous system. However, behavioral methods of assessing peripheral nerve regeneration are still lacking. In the present work we used the median nerve and finger flexors as the experimental model. After resection or crush injury of the median nerve, rats were submitted daily to the grasping test. For this, they were gently lifted by the tail and allowed to grasp a grid connected to an ordinary electronic balance. While grasping the animal continued to be lifted by the tail with increasing firmness until it lost its grip. At this precise moment the value shown by the balance was recorded. In the crushed median nerve, recovery of function was clearly demonstrated by the grasping test. The test also indicated the exact day on which recovery began and its improvement with time. This very simple objective behavioral method provides a sensitive quantitative technique for assessing recovery. The experimental model using the median nerve seems to be better than the sciatic nerve model, as no articular contractures or autotomies were noted in the present experimental animals. PMID- 7475221 TI - Computer analysis and quantification of periaqueductal grey-induced defence behaviour. AB - The pharmacological study of midbrain-evoked aversive behaviours is commonly measured in terms of electrical escape thresholds, although studies which examine the actual expression of the behaviour are being introduced. A computer-driven automated tracking system has been developed to record and analyse the very rapid locomotor activity produced by the defence response. Stimulation of the midbrain periaqueductal grey (PAG) matter in rats with the excitatory amino acid D,L homocysteic acid (DLH) produced very rapid and short-lasting escape behaviour which was tracked by computer. The system, VideoTrack, determines the speed and distance travelled by the animal and stores and statistically analyses the data. VideoTrack was able to detect a pharmacological modifications of the response; anti-aversive effects produced by intra-PAG pretreatment with the 5-HT1A agonist 8-hydroxy 2-(di-n-propylamino)tetralin (8-OHDPAT) and its reversal by peripheral pretreatment with the 5-HT1A antagonist N-[2-[-(2-methoxyphenyl)-1 piperazinyl]ethyl-N- (pyridinyl)cyclohexanecarboxanide (WAY 100635). PMID- 7475222 TI - Photolysis of a newly synthesized caged glycine activates the glycine receptor of rat CNS neurons. AB - The functions of 12 different newly synthesized caged glycines were examined on strychnine-sensitive glycine receptors on rat hypothalamic neurons using photochemical and physiological methods. Quantitative analysis of Cl- current induced by photolyzed glycines revealed N-[1-(2-nitrophenyl)ethylcarbonoxyl] glycine (N4-caged glycine) to be the most effectively photolyzed. The current amplitude induced by photolyzed N4-caged glycine was almost the same as that induced by normal glycine. We propose N-[1-(2-nitrophenyl)ethylcarbonoxyl] to be a useful photolabile moiety for new caged neurotransmitters. PMID- 7475223 TI - Measurement of vascular permeability in spinal cord using Evans Blue spectrophotometry and correction for turbidity. AB - Vascular permeability can be visualized by Evans Blue (EB) extravasation and quantified by spectrophotometry after formamide extraction of the tissue. However, formamide extracts show significant turbidity, which may contribute to the total optical density at the wavelength of measurement (e.g., 620 lambda). We developed a simple method for estimating the component of the total optical density of a dyed specimen contributed by turbidity. Our method, which uses a determination of turbidity made at another point of the light spectrum (740 lambda), was more precise than two other EB quantification techniques. We therefore recommend it for individual correction of formamide extracts of spinal cord specimens. The application of this technique to the brain remains to be determined. PMID- 7475224 TI - A method allowing intracellular and extracellular single-unit recordings from brain slices in the grease-gap chamber. AB - The grease-gap chamber makes it possible to perfuse two parts of a brain slice separately. Hence, it lies between bath application and iontophoresis or pressure ejection in terms of the size of the region that receives controlled drug dosing. Modifications of a commercially available chamber are described which have permitted the first extracellular and intracellular single-unit recordings to be taken from brain slices in a grease-gap preparation. In addition, we describe the electrical resistance of the gap as a measure of the integrity of the barrier and a simple method for monitoring this resistance continuously. The resistance monitor is particularly useful during low flow rate conditions that improve mechanical stability. These techniques extend the grease-gap method to electrophysiological studies of single cells in slices. PMID- 7475225 TI - A polyclonal antibody against mammalian FOS can be used as a cytoplasmic neuronal activity marker in a teleost fish. AB - A polyclonal antibody raised against a conserved region of a mammalian FOS sequence was tested for its use as an activity marker in the rainbow trout. The FOS-like expression in the trout is entirely cytoplasmic and appears in a Nissl like pattern. The reaction is specifically induced by both orthodromic and antidromic electrical stimuli and during motor responses evoked by natural stimulation, although some positive neurons are found at locations that are not obviously related to the presented stimuli. Following spinal nerve stimulation, antidromically activated motoneurons were found to be positive in the ipsilateral spinal cord. Orthodromic driving of spinal moto- and interneurons by stimulation of the medial longitudinal fasciculus (MLF) in the hindbrain evoked FOS-like immunoreactivity throughout the motor column in the spinal cord, but not in regions lying caudal to a lesion of the MLF-axons. Evoking about 25 startle responses by natural auditory stimulation gives FOS-like immunoreactivity in the Mauthner cell, which initiates the response, whereas positive Mauthner cells were never observed in control fish. The stimulation protocols that were used strongly activated the stimulated cells and so the observed FOS-like immunoreactivity might be related to an increase protein synthesis needed to restore their depleted transmitter levels. PMID- 7475226 TI - A direct comparison of GFAP immunocytochemistry and GFAP concentration in various regions of ethanol-fixed rat and mouse brain. AB - Glial fibrillary acidic protein (GFAP) immunostaining is the most commonly used method to examine the distribution of astrocytes and the hypertrophy of astrocytes in response to neural degeneration or injury. More recently, a variety of biochemical assays for GFAP have been developed. Both qualitative immunocytochemical evaluations of GFAP and quantitative biochemical measurements of GFAP have been used to examine the regional distribution of GFAP within the central nervous system (CNS). The former method has largely been based on aldehyde-fixed tissue, while the latter approach has been based on the use of fresh tissue extracts or homogenates. In the present study, we used ethanol as a fixative to permit both immunocytochemical and biochemical procedure to be carried out on brain tissue from a single animal. Normal adult rats and mice were perfused with a 70% ethanol/saline solution, and each brain was hemisectioned. The concentration of GFAP was measured in regions of 1 hemisection, using an enzyme-linked immunosorbent assay (ELISA), while the other hemisection was used for GFAP immunostaining. Regional differences occurred in the brains of both species, with the highest concentration of GFAP found in the brainstem, and the lowest concentrations found in the striatum and cortex. The specific patterns of GFAP immunoreactivity corresponded to regional concentrations in most brain area of both species. These data show that it is possible to assay GFAP concentrations in tissue prepared for immunocytochemical analysis, providing both qualitative and quantitative information from one set of tissue. PMID- 7475227 TI - Controlled superfusion of the rat spinal cord for studying non-synaptic transmission: an autoradiographic analysis. AB - Recently, evidence has been raised that long-term changes in the central nervous system are mediated by extrasynaptic spread of neuropeptides ('volume transmission'). To study the effects of volume transmission in the spinal cord we developed the technique of controlled superfusion of the rat cord dorsum. This paper presents quantitative data about the spread, local spinal tissue concentration and redistribution of (2-[125I]iodohistidyl)neurokinin applied for 15, 30 or 60 min to the spinal cord dorsum in concentrations of 0.05 or 50 microM (10 microliters). Analysis of autoradiograms of sagittal and transverse spinal cord sections was done by computer-assisted densitometry. Under all experimental conditions, the spread of radiolabel into the superfused spinal cord segments reached Rexed's laminae V and VI; maximal spread (1.6 +/- 0.3 mm) was measured after superfusion for 30 min. The amount of radiolabel decreased in ventral direction as a function of distance. Highest tissue concentrations of neurokinin A (NKA) were obtained within the superficial spinal cord up to a depth of 0.5 mm and ranged from 700 to 2000 pmol/g following superfusions for 15 or 30 min with 50 microM NKA. Thus, these tissue concentrations were 25-70 times lower than the concentration of NKA in the superfusate. Since pool content was not exchanged, the radioactivity within the spinal cord was lower after superfusion periods of 60 min than after 15 or 30 min. Detection of radiolabel in blood and urine suggests that capillary clearance is relevant and limits the accumulation of the peptide within the spinal cord tissue and the spread into deeper laminae. The controlled superfusion of the rat cord dorsum is a useful method to mimick the spinal release of endogenous neuropeptides such as NKA during intense noxious stimulation, and it can be employed for versatile investigations of the effects of neuroactive molecules on the processing of sensory information in the intact spinal network. PMID- 7475228 TI - A new interface chamber for the study of mammalian nervous tissue slices. AB - We describe a new interface-type chamber for electrophysiological studies in mammalian brain slices. Thermoregulation of the inner recording chamber is achieved using the Peltier effect and a feedback control unit. Between 15 and 40 degrees C, and for perfusion rates from 1 to 5 ml/min, the temperature can be maintained within +/- 0.1 degrees C of the command value; it can also be rapidly and reliably changed. An external bath, heated by a coiled resistor, generates a humidified, oxygenated atmosphere diffusing above the slices. Survival of neuronal tissue is excellent and stable intracellular recordings can be obtained using either sharp or patch-clamp micropipettes. Perfusion solutions can be readily exchanged, rendering this chamber suitable for the study of bath-applied neuroactive compounds. PMID- 7475229 TI - Multi-electrode array for measuring evoked potentials from surface of ferret primary auditory cortex. AB - Using silicon-integrated circuit technology, we have fabricated a flexible multi electrode array and used it for measuring evoked potentials at the surface of the ferret primary auditory cortex (AI). Traditionally, maps of cortical activity are recorded from numerous sequential penetrations with a single electrode. A common problem with this approach is that the state of the cortex (defined in part by level of anesthesia and number of active cells) changes during the time required to generate these maps. The multi-electrode array reduces this problem by allowing the recording of 24 locations simultaneously. The specific array described in this report is designed to record cortical activity over a 1 mm2 area. It is comprised of 24 gold electrodes (40 x 40 microns2) each spaced 210 microns apart. These electrodes are connected to contact pads via gold leads (5 cm in length). The electrodes, leads, and contact pads are sandwiched between two layers of polyimide. The polyimide passivates the device and makes the device flexible enough to conform to the shape of the cortex. The fabrication procedures described here allow various other layouts and areas to be readily implemented. Measurements of the electrical properties of the electrodes, together with details of the multichannel amplification, acquisition, and display of the data are also discussed. Finally, results of AI mapping experiments with these arrays are illustrated. PMID- 7475230 TI - A simple, rapid HPLC method for the concurrent measurement of cocaine and catecholamines in brain tissue samples. AB - We describe a simplified HPLC method for the measurement of cocaine and catecholamines in the same brain tissue sample. Using this method to measure cocaine in the brain of rats which received either 10 or 20 mg/kg cocaine injections, we found substantial cocaine concentration differences between medial prefrontal cortex, striatum and limbic brain areas. Specifically, for each cocaine dose level there was a nearly 2-fold differential between cocaine concentration in the cortex as compared to limbic tissue. A shortcoming of many neurobehavioral studies of cocaine effects is the absence of brain cocaine measurements. The simplicity of the present method makes the measurement of cocaine and catecholamines from the same tissue sample a procedure which can readily be incorporated in the studies designed to evaluate the neurochemical effects of cocaine treatments. PMID- 7475231 TI - Interpreting nerve morphometry data after experimental traumatic lesions. AB - Morphometric data were obtained in 2 experimental conditions: crush lesions of the sciatic nerve of rats, and transection of this nerve followed by repair. Ratios were used in order to facilitate comprehension of data. Results were compared to those obtained using a method of functional assessment. The study indicated that morphometric data provided a reliable picture of the trophic condition of regenerated nerves but did not correlate with the level of function. PMID- 7475232 TI - A spring-activated tilting apparatus for the study of balance control in man. AB - A tilting apparatus has been designed and constructed to produce fore-aft whole head-and-body tilt (WHBT) alone, or in combination with ankle dorsiflexion of standing subjects, about an axis colinear with the ankle joint. The apparatus is composed of a vertical tilting structure attached to a supporting base. Mechanical rotation of the vertical tilting structure is achieved by a spring activating system mounted on its base. Subjects are secured to the vertical structure by a body harness system with the head fixed, and the feet secured to the standing platform. Simultaneous dynamic WHBT and ankle joint displacement are induced by rotating the vertical support; whereas WHBT alone is achieved by concurrent rotation of both the vertical support and standing platform. Tilts are triggered manually, and data acquisition precedes head acceleration onset by 50 100 ms to provide suitable baseline values. This tilting apparatus has been successfully used to apply forward WHBT in 34 subjects with height ranging from 1.55 to 1.87 m and weights from 42 to 95 kg, and at magnitudes of peak head acceleration varying from 0.4 to 2.2 g as measured by a linear accelerometer mounted on a dental bite. These acceleration rates can be reproduced with minor variation in the same subjects. Moreover, the area under the head acceleration traces was reproducible within 15% between subjects. Tilts can be delivered with concomitant ankle dorsiflexion. These features make the low-cost tilting apparatus a very useful tool for the study of human tilting reactions in both laboratories and clinical settings. PMID- 7475233 TI - Evaluation of single-channel gating kinetics produced after amplitude-based separation of unitary currents. AB - Cell-attached patch-clamp recording has been established as a major technique for investigating ion channel behaviour in a physiological setting, despite the problems which arise in analysing records containing more than one type of unitary current. To circumvent these problems, single-channel amplitude-based assignment of discrete single-channel events to different channel types becomes increasingly necessary. Surprisingly, a systematic evaluation of the validity of this method in determining single-channel parameters has not been performed to date. Using computer-stimulated single-channel traces, and recordings from a biological preparation containing well-characterized ion channels (N1E-115 neuroblastoma cells), we have explored the accuracy by which amplitude-based separation recovers ion-channel parameters. Determination of gating kinetics after separation revealed that even a very small contamination in the selected population yields additional time constants in the probability density functions. Therefore, our results demonstrate that, whereas the use of amplitude-based separation is straightforward for determining slope conductance and reversal potential, it is prone to incorporate errors in establishing gating kinetics. Ways of identifying such errors are described. PMID- 7475234 TI - Use of BCECF and propidium iodide to assess membrane integrity of acutely isolated CA1 neurons from rat hippocampus. AB - We used 2 fluorescent dyes, 2',7'-bis(carboxyethyl)-5(6)-carboxyfluorescein (BCECF) and propidium iodide (PI), to assess the membrane integrity of neurons acutely isolated from the CA1 region of the rat hippocampus. Exciting BCECF at a relatively pH-insensitive wavelength (440 nm), or exciting PI at 490 nm, we quantitatively recorded, in real time and in single cells, the rate constants for BCECF loss (-k440) and PI uptake (k490). We found that approximately 98% of intracellular BCECF is rapidly released by applying 0.01% saponin. In neurons not treated with saponin, rate constants for BCECF loss and PI uptake typically were 1% min-1 or less under control conditions, in the presence of NH3/NH4+ and in the absence of Na+. However, in a small number of neurons, the rate constant for BCECF loss increased markedly (-k440 > 5% min-1), while pHi approached pHo, suggesting that the plasma membrane spontaneously became leaky. When neurons were progressively swollen in hypotonic solutions, rates constants for BCECF loss and PI uptake generally were affected minimally unless osmolality was decreased to approximately 75 mOsmol/kg. Treating neurons with 0.001% saponin caused an increase in PI uptake rate only in a minority of neurons, whereas in most experiments a similar treatment caused -k440 for BCECF to exceed 5% min-1, and led to a rapid deterioration of the pH gradient across the cell membrane. At even lower saponin levels (0.0005-0.0007%), we observed a much slower deterioration of pHi, which occurred at low rates of BCECF loss (-k440 = approximately 3% min-1). We conclude that computing rate constants for BCECF loss and PI uptake may be useful for assessing neuronal health, and that BCECF loss may be more sensitive to cell damage than PI uptake. PMID- 7475235 TI - An improved protocol for immunogold staining for scanning electron microscopy of cultured cells. AB - This paper describes an effective protocol for preparing immunogold-labelled cultured cells for high-resolution scanning electron microscopy. The cells examined were rat peripheral nerve Schwann cells and fibroblasts. The protocol employs low concentrations of chemicals and decreased fixation time. The resultant immunohistochemical cell labelling has little or no background staining, while maintaining the morphological integrity of the cell membrane. PMID- 7475236 TI - Investigating dynamic aspects of brain function in slice preparations: spatiotemporal stimulus patterns generated with an easy-to-build multi-electrode array. AB - Electrical stimulation of nervous tissue with single stimulating electrodes is a technique widely used for the investigation of nervous system function. While it has proved to be useful in all kinds of experiments, single electrode stimuli are, however, far from being 'natural'. In most parts of the living brain, incoming activity results from the firing of a large number of presynaptic neurons, thus reflecting a complex combination of space and time aspects of neural activity. In this paper, a multi-electrode stimulating system is introduced which allows for the generation of fast space-time stimulus patterns. An example for the application of dynamic input patterns to the cerebellar cortex in vitro is given. The corresponding experiments revealed aspects of cerebellar function which cannot be seen using static or single electrode stimulation. PMID- 7475237 TI - Evaluation of a video tracking device for measurement of horizontal and vertical eye rotations during locomotion. AB - We have evaluated a video-based method for measuring binocular horizontal and vertical eye movements of human subjects by comparing it with the magnetic search coil technique. This video tracking system (VTS) uses multiple infrared light sources and small video cameras to simultaneously measure the positions of reflected corneal images and the center of the pupil. The system has a linear range of approximately +/- 40 degrees horizontally and +/- 30 degrees vertically, a sampling rate of 120 Hz (180 Hz with the head fixed), and system noise with standard deviation of < 0.04 degree. The binocular eye-tracking system is light weight (190 g), being mounted on goggles that, with the eyes in primary position, permit a field of view of 60 degrees horizontally and vertically. The VTS is insensitive to translations of the tracker relative to the eyes. By placing the video preprocessing unit on a cart, eye movements may be recorded while subjects walk through distances up to 100 feet. In comparison with the magnetic search coil technique, the VTS generally provides reliable measurements of horizontal and vertical eye position; eye velocity is noisier than corresponding coil signals, but superior to electro-oculography. PMID- 7475238 TI - N-(2-chloroethyl)-N-ethyl-2-bromobenzylamine (DSP-4) has differential efficacy for causing central noradrenergic lesions in two different rat strains: comparison between Long-Evans and Sprague-Dawley rats. AB - We tested the hypothesis that Long-Evans (LE) and Sprague-Dawley (SD) rat strains were equally sensitive to the noradrenergic neurotoxin N-(2-chloroethyl)-N-ethyl 2-bromobenzylamine (DSP-4) with respect to central lesions of locus coeruleus (LC) terminals as measured by immunohistochemical localization of dopamine-beta hydroxylase (D beta H). Analysis of D beta H immunoreactivity was made by both qualitative and quantitative methods. Intraperitoneal injections of 50 mg/kg DSP 4 caused a dramatic reduction of noradrenergic terminals in the neocortex, hippocampus and cerebellum of SD, but not LE rats as compared to saline-injected controls. This finding indicates that LE rats are less sensitive than SD rats to the neurotoxic effects of DSP-4 in the central nervous system. PMID- 7475239 TI - Second-messenger responses in brain slices to elucidate novel glutamate receptors. AB - G-Protein-coupled or 'metabotropic' glutamate receptors (mGluRs) are a novel heterogenous family of excitatory amino acid receptors. Activation of mGluRs in the rat hippocampus by the mGluR-selective agonist 1S,3R-1-aminocyclopentane-1,3 dicarboxylic acid (1S,3R-ACPD) leads to multiple changes in second-messenger formation. These include increases in basal phosphoinositide hydrolysis, decreases in forskolin-stimulated cAMP formation, and enhancement of cAMP formation via a potentiation of the effects of endogenous adenosine. These changes in mGluR coupling to phosphoinositide hydrolysis and the formation of cAMP likely reflect the in situ expression of heterogenous populations of mGluRs. A number of electrophysiological studies on the functions of mGluRs in hippocampal circuitry, ontogeny, and cellular functions have also been described. Any or all of these mGluR-mediated changes in second messengers may underlie the reported cellular effects associated with the mGluR activation by 1S,3R-ACPD. However, mGluR agonists that have selectivity for different mGluR second messenger pathways are needed to sort out the cellular consequences of activating in situ expressed mGluR subtypes linked to specific second-messenger pathways. PMID- 7475240 TI - Synaptic plasticity in the hippocampal slice: functional consequences. AB - There are 3 known forms of synaptic plasticity at CNS synapses: long-term potentiation (LTP) mediated by NMDA receptor activation, LTP mediated by voltage dependent calcium channel (VDCC) activation, and long-term depression (LTD) mediated by the NMDA receptor. All 3 forms of synaptic plasticity can be observed in hippocampal CAl cells, all are induced by afferent activation, all involve Ca2+ influx, and all activate Ca(2+)-dependent mechanisms. We consider the functional consequences of the presence of 3, sometime opposing, forms of synaptic plasticity at the same synapse. We suggest that the 2 forms of LTP have different consequences for the synapse. We postulate that the co-existence of potentiating and depressing capabilities influences the network processing capabilities of neural networks. PMID- 7475241 TI - Effects of K+, pH and glutamate on 45Ca kinetics in hippocampal brain slices. AB - Altered calcium homeostasis is likely to play a pathogenetic role in cerebral ischemia. In order to further understand which factors associated with ischemia contribute to disturbances of calcium metabolism, the influence of 3 isolated insults, 8 mM K+, pH 6.1 and 1 mM glutamate, on total tissue calcium were studied by analysis of steady-state kinetics of 45Ca in 500 microns hippocampal brain slices. 45Ca kinetics were analyzed with 2 bi-exponential models by non-linear least-squares analysis. Tissue wet weight/protein was measured simultaneously. Each experimental condition produced a unique tissue response. Raising K+ had no effect on tissue water but increased the rate of uptake of Ca2+ into the larger, rapidly equilibrating tissue Ca2+ space. Acidosis reduced tissue water and the amount of Ca2+ in the slowly equilibrating compartment due to enhanced efflux from that space. Glutamate increased tissue water in a time-dependent manner and increased the influx and amount of Ca2+ in the slowly equilibrating space. Combined insults revealed minimal interaction between K+ and acidosis or glutamate, but glutamate with acidosis worsened tissue injury. We discuss the relationship of this technique to other methods for studying tissue calcium and the significance of the observations regarding ischemia. PMID- 7475242 TI - Hippocampal slices: glutamate overflow and cellular damage from ischemia are reduced by sodium-channel blockade. AB - We evaluated concentrations of excitatory amino acids released from slices into the superfusing solution and also evaluated extracellular field potential recordings and histological appearance of slice tissues to evaluate several sodium-channel modulating drugs as potential treatments for ischemia. The selective sodium-channel blocker tetrodotoxin (TTX, 1 microM) reduced glutamate release from deprivation of oxygen and D-glucose, while calcium-channel blockade was ineffective. Thus, during ischemia, we propose that glutamate may be released from the free cytosolic pool ('metabolic' glutamate) rather than by exocytosis. TTX (100-500 nM) and voltage-dependent sodium-channel blockers (phenytoin, 20-100 microM; lidocaine, 2-200 microM) each prevented damage to slices without blocking action potentials. The reduction of cellular depolarization and sodium loading during ischemia may explain the neuroprotective action of several sodium-channel modulating drugs in our in vitro studies and also in animal models. PMID- 7475244 TI - Preparative methods for brain slices: a discussion. AB - Criteria for slice health and factors that affect slice health were discussed by many of the participants in the conference. In addition to the standard parameters of slice health (energy metabolism, morphology, electrophysiological responsiveness) more subtle but possibly equally important manifestations of slice health were discussed. These included protein synthesis, and more subtle changes, of which we are becoming increasingly aware. The latter include synthesis of stress-related proteins, altered levels of phosphorylation, altered levels of proteolysis. These last were only touched on, but it is becoming apparent they do in fact constitute important manifestations of differences between the slice preparation and the in vivo tissue. They may well lead to quite different responses in slices from those that occur in vivo. While many ways of optimizing slice wellness were discussed, there was a fair consensus that certain adjustments will optimize the most widely measured aspects of cell function. These include the following, wherever possible. Use of young animals, use of the interface chamber, preparing slices with the vibratome, pre-treating animals with ice-cold cardiac perfusion before sacrificing, using pre-incubation media which reduce NMDA receptor activation, free radical formation and cell swelling. When possible these treatments should perhaps be continued into the normal incubation. This being said, many viewpoints were actually expressed in the discussion, and it should be read to get a feel for the usefulness of the different approaches. PMID- 7475243 TI - Hypoxia, excitotoxicity, and neuroprotection in the hippocampal slice preparation. AB - The excitotoxic hypothesis postulates a central role for the excitatory amino acids (EAAs) and their receptors in the neuronal damage that ensues cerebral ischemia-hypoxia and numerous other brain disorders. A major premise of the excitotoxic hypothesis is that neuronal protection can be achieved via blockade of EAA receptors with specific antagonists. This paper describes the use of the rat hippocampal slice preparation in the evaluation of various EAAs and their analogues for their potency as excitotoxins (agonists) and antagonists of the NMDA and the kainate/AMPA glutamate receptor subtypes. The hypersensitivity of hypoxic hippocampal slices to the presence of excitotoxins provided us with an inexpensive, sensitive tool to distinguish between structurally similar compounds. Moreover, these studies indicate that hypoxic neuronal damage cannot solely result from an excitotoxic mechanism; the involvement of voltage-dependent calcium channels in such damage is likely, as is evident from experiments performed in calcium-depleted medium and with the non-competitive NMDA antagonist MK-801. At sub-toxic doses, quinolinate, a tryptophan metabolite implicated in Huntington's disease, appears to be a strong potentiator of the toxicity of all excitotoxins tested. PMID- 7475245 TI - Making the best of brain slices: comparing preparative methods. PMID- 7475246 TI - Studies on the role of metabotropic glutamate receptors in long-term potentiation: some methodological considerations. AB - There has been considerable interest recently in trying to elucidate the roles of metabotropic glutamate receptors (mGluRs) in the induction of long-term potentiation (LTP) in area CA1 of rat hippocampal slices. This has come about principally because of the development of specific mGluR agonists and antagonists. Recently we reported that the competitive mGluR antagonist (+)-alpha methyl-4-carboxyphenylglycine (MCPG) blocks the induction of LTP but not short term potentiation (STP). We describe here the dose-dependency of the MCPG block; there is no effect at 100 microM while at 200 microM the block of LTP is normally complete but STP is spared. A higher concentration of MCPG (500 microM) has the same effect as 200 microM. We have also reported recently that high-frequency (tetanic) stimulation conditions a pathway such that MCPG fails to block the induction of subsequent LTP. We illustrate here that the conditioning effect of a tetanus lasts at least 6 h. We show how the pathway can be conditioned, without any persistent change in the synaptic response, by delivering tetanic stimulation in the presence of the specific NMDA receptor antagonist D-2-amino-5 phosphonopentanoate (AP5). The pathway can subsequently be deconditioned by delivering low-frequency stimulation (900 shocks at 2 Hz) so that MCPG blocks the induction of subsequent LTP. We also have reported that the specific mGluR agonist 1-aminocyclopentane-(1S,3R)-dicarboxylate (ACPD) can induce LTP without the need for STP.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7475247 TI - Imaging of calcium inside dendrites and spines: goals, issues and perspectives. AB - The growing interest in dendritic spines in recent years originates from both the realization that the spine is likely to be the site where long-term plastic changes in synaptic properties take place, and that imaging methods are available which allow, for the first time, the study of these changes in the living dendritic spine. This report briefly summarizes methodological and biological issues associated with the study of dendritic spines in living tissue. The combined use of electrical and imaging methods for the study of dendritic spines certainly will contribute to a better understanding of synaptic integration and plasticity. PMID- 7475248 TI - The use of brain slices and dissociated neurones to explore the multiplicity of actions of 5-HT in the central nervous system. AB - The cellular actions of 5-hydroxytryptamine (5-HT) on adult and neonatal rat central neurones have been investigated in detail using a combination of in vitro slice and dissociated neurone preparations. Patch-clamp recordings from acutely dissociated adult rat dorsal raphe neurones confirm data obtained using conventional slice preparations that 5-HT activates an inwardly rectifying potassium channel through a 5-HT1A receptor leading to hyperpolarization of the cell. Single-channel recordings indicate that this pathway requires only the involvement of a pertussis toxin-sensitive G-protein. Adult rat facial motoneurones in conventional slices are depolarized by 5-HT through a combination of mechanisms, closure of potassium channels and enhancement of the hyperpolarization-activated, cationic current, IH. Distinct receptors appear to mediate these two actions. Both mechanisms are present in visually indentified neonatal rat facial motoneurones in thin brain slices. Whole-cell patch-clamp recordings show the action of 5-HT on IH to mediate a caesium-sensitive inward current which can be mimicked by the adenylate cyclase activator, forskolin. The experimental data illustrate how a range of complimentary in vitro electrophysiological techniques can be employed to unravel neurotransmitter mechanisms and pharmacology. PMID- 7475249 TI - Looking for chaos in brain slices. AB - Many signals measured from the nervous system exhibit apparently random variability that is usually considered to be noise. The development of chaos theory has revealed that such random appearing variability may not, in fact, be random, but rather may be deterministic behavior that can reveal important information about the system's underlying mechanisms. We present some new methods for distinguishing determinism from randomness in experimental data, and we apply these methods to population neural responses recorded from hippocampal tissue slices. PMID- 7475251 TI - The brain slice preparation: a tribute to the pioneer Henry McIlwain. AB - Henry McIlwain is usually remembered for his major contributions to the discipline of neurochemistry. He also had a profound influence on electrophysiology by inventing and establishing the brain slice technique. This article describes some of his pioneering studies where he devised ways of preparing, both manually and mechanically (using the McIlwain tissue chopper), viable slices of mammalian cortical tissue and showed how these could be maintained in a brain slice chamber. His work with Choh-Luh Li (Li and McIlwain, 1957) on the first intracellular recording from brain slices and his work with Chosaburo Yamamoto (Yamamoto and McIlwain, 1966) describing the first study of synaptic transmission in brain slices are highlighted. In these studies, McIlwain started the use of brain slices to investigate functional anatomy, to study the effects of hypoxia and hypoglycaemia, to perform quantitative pharmacological analyses and to study synaptic plasticity: investigations which continue today to provide important insights into the functioning of the brain in health and disease. PMID- 7475250 TI - Neurophysiology of neocortical slices resected from children undergoing surgical treatment for epilepsy. AB - The recent emergence of surgical treatment of childhood epilepsy has led to the accessibility of young human cerebral tissue for electrophysiological studies of the mechanisms involved in epileptogenesis. Intracellular recordings were obtained from neurons in slices prepared from neocortical tissue resected from children (3 months to 15 years) with catastrophic epilepsy. Data from 'least abnormal' versus 'most abnormal' tissue were compared; the evaluation of the degree of abnormality was based on several clinical criteria. Hypotheses concerning NMDA receptors, local synaptic circuits, and epileptiform bursts were tested. The NMDA receptor-mediated component of synaptic responses, which was isolated pharmacologically, had a voltage dependence that was functionally mature by 8-10 months of age and did not appear to be altered even in the most abnormal tissue. Local inhibitory and excitatory synaptic circuits were present as early as 11 months and 8 months, respectively. Local excitatory circuits were sufficiently extensive in young children to initiate and sustain epileptiform activity when synaptic inhibition was suppressed. Bicuculline-induced epileptiform bursts were similar to those in adult human or animal neocortical slices. Burst duration and the presence of after-discharges were unrelated to patient age or tissue abnormality. These data demonstrate that (1) the electrophysiological properties of human neocortical neurons are very similar to those observed in animal experiments, (2) the mechanisms of neuronal communication are qualitatively mature within the first year of life, and (3) synaptic transmission and local neuronal circuits appear qualitatively normal, even in the most abnormal tissue from children with catastrophic epilepsy. PMID- 7475252 TI - Patch-clamping in brain slices: synaptic transmission from ATP to long-term potentiation. AB - Application of patch-clamp techniques to brain slices has resulted in an enormous increase in the resolution of synaptic currents in mammalian central neurones. This improved resolution has allowed direct observation of miniature and evoked synaptic currents, leading, in agreement with other findings, to the conclusion that the quantal size of synaptic currents in the brain is limited by the number of postsynaptic receptors. Possible explanations for the skewed miniature distribution, observed at all fast central synapses are discussed, with reference to anatomical observations. As a result, a model is proposed which is consistent with much of the apparently contradictory data on the induction and maintenance of long-term potentiation. In addition to the study of mechanisms of synaptic transmission, improved resolution provided by the patch-clamp technique has allowed resolution of synaptic currents, in the brain, mediated by ATP. The role of ATP as a central neurotransmitter is discussed in terms of the above findings. PMID- 7475253 TI - Use of the whole-cell patch-clamp method in studies on the role of cAMP in regulating the spontaneous firing of locus coeruleus neurons. AB - The whole-cell patch-clamp technique represents a major advance over conventional intracellular recordings in the study of the modulation of ion channels by intracellular messengers. This report illustrates how application of the whole cell technique to noradrenergic neurons of the rat locus coeruleus in brain slices has led to the finding that cAMP via its phosphorylation pathway modulates tonic pacemaking in these neurons. In the studies to be described, the particular advantage of the whole-cell technique was that it allowed introduction of macromolecules related to the cAMP pathway (e.g., protein kinase inhibitor and protein kinase A) directly into cells. Furthermore, these studies were carried out in situ, in thick brain slices allowing a direct comparison with a large body of existing extracellular and intracellular data obtained under similar conditions. PMID- 7475254 TI - Single-channel and whole-cell studies of calcium currents in young and aged rat hippocampal slice neurons. AB - The hippocampal slice preparation has contributed greatly to analysis of the basic neurophysiology of brain neurons. In addition, because traumatic dissociative procedures are not used, the in vitro slice is particularly well suited for studies of electrophysiological properties of hippocampal neurons in young and aged rodent brain. Using the slice, we have previously observed an aging-dependent enhancement of voltage-activated Ca2+ influx using a combination of intracellular sharp electrode current-clamp and voltage-clamp techniques. The Ca(2+)-dependent afterhyperpolarization as well as the Ca2+ action potential were significantly larger in aged rat neurons. Using the sharp electrode clamp method, similar effects were found for high voltage-activated whole-cell Ca2+ currents. In order to study the mechanistic bases of these aging phenomena at the single channel level, we have recently focused on recording cell-attached patches from neurons in the partially dissociated hippocampal slice ('zipper' slice). This technique, developed by Gray et al. in 1990, subjects slices to a mild enzymatic treatment resulting in the exposure of individual neurons for patch-clamp procedures. Using this technique, we are currently recording single Ca2+ channel activity in hippocampal slices from 4- to 29-month-old rats. PMID- 7475255 TI - Immunocytochemical and in situ hybridization approaches to the optimization of brain slice preparations. AB - Methods are described for determining the expression of specific mRNAs and proteins in brain slices, in order to elucidate changes in gene expression during preparation of vibratome slices from hippocampus of adult rats. In situ hybridization with 35S-labeled oligonucleotides was used to evaluate the level and distribution of c-fos and hsp72 mRNAs in 15-microns frozen sections prepared from these slices. Commercially available antibodies were used to examine the distribution of induced Fos and Jun proto-oncogenes as well as expression of the neuronal cytoskeletal protein, microtubule-associated protein 2 (MAP2), in 50 microns vibratome sections from immersion-fixed slices. These studies confirm the induction of c-fos and hsp72 mRNAs during routine incubation, as previously observed in hippocampal slices obtained with a tissue chopper and incubated under somewhat different conditions, indicating that such responses are likely to be common features of many slice preparations. Accumulation of Fos and Jun immunoreactivities in neurons and glia was generally consistent with the distribution of c-fos mRNA induction observed in slices, and the neuronal component of this response was comparable to the expression of these proteins observed after transient ischemia in vivo. MAP2 immunoreactivity detected in the dendritic processes of neurons tended to show an increase in staining intensity during slice incubation, although loss of dendritic staining in specific regions was occasionally observed in association with the absence of Fos and Jun expression and histological evidence of neuron damage. These results support the use of MAP2 immunoreactivity as a sensitive indicator of neuronal integrity in slices.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7475256 TI - Use of brain slices in the study of free-radical actions. AB - To understand the neuropathological roles of free radicals we investigate their actions in a model neuronal system, the hippocampal brain slice. Free radicals can be generated through a number of methods: hydrogen peroxide to produce hydroxyl radicals, dihydroxyfumarate to generate superoxide and ionizing radiation producing a variety of radical species. We find that free radicals have a number of profound effects in this system, which can be prevented by free radical scavengers and antioxidants. With exposure to free radicals, the ability to generate spikes and synaptic efficacy are impaired. Decreased spike generating ability is correlated with lipid peroxidation. No change in membrane potential, membrane resistance, or many of the potassium currents can account for the effect on spike generation. Protein oxidation is likely to underlie synaptic damage. Both inhibitory and excitatory synaptic potentials are reduced by free-radical exposure. Presynaptic mechanisms are implicated. Lower concentrations of radicals prevent the maintenance of long-term potentiation, perhaps through oxidation of the NMDA receptor. The actions of the free radicals are often reversible because of the presence of repair mechanisms, such as glutathione, in hippocampal slices. The brain slice preparation has allowed us to begin to understand the electrophysiological and biochemical consequences of free-radical exposure. PMID- 7475257 TI - Pathophysiology of anoxic depolarization: new findings and a working hypothesis. AB - Anoxic depolarization has been linked to the generation of hypoxic irreversible damage. Treatments that postpone its occurrence during hypoxia protect against irreversible damage. This work investigates possible mechanisms leading to anoxic depolarization and ways to prevent or delay it. Exogenous creatine (a compound that delays ATP depletion during hypoxia by increasing the intracellular store of phosphocreatine) doubles the latency of anoxic depolarization. Ouabain (100 microM) reproduces in normoxic slices the depolarization of anoxic depolarization and the concurrent changes in [K+]0; thus, failure of (Na+, K+)ATPase (which is likely to occur during hypoxia due to ATP depletion) is sufficient to cause anoxic depolarization. Electrophysiological evidence, however, suggests that failure of this ATPase causes anoxic depolarization through some intermediate event, probably Na(+)-induced cell swelling. In accordance with this hypothesis, increasing extracellular osmolarity with mannitol (25 mM) increases anoxic depolarization latency by approximately 25%. Other possible mechanisms of anoxic depolarization are also discussed. PMID- 7475258 TI - Lymphatic microsurgery: a potent weapon in the war on lymphedema. PMID- 7475259 TI - The diagnosis and treatment of peripheral lymphedema. Consensus document of the International Society of Lymphology Executive Committee. PMID- 7475260 TI - Benign supraclavicular tumorous lymphangiectasia--a new disease? AB - We describe an isolated recurrent non-inflammatory tumorous swelling of the supraclavicular fossa in four premenopausal women. Ultrasonography, magnetic resonance imaging and computer tomography of the neck each suggested an inhomogeneous mass consistent with "lymphangioma." In each patient the clinical course and histopathologic findings suggested that the swellings were due to chronic localized lymph stasis with subsequent lymphangiectasia, possibly initiated by intermittent obstruction of the juncture of the thoracic or right lymph duct with the internal jugular vein. Enlargement may have been hormonally triggered by estrogens as each woman was taking oral contraceptive pills at the onset of the disease. To characterize this unique entity, we have termed the disorder benign supraclavicular tumorous lymphangiectasia. PMID- 7475261 TI - Correlation between AIDS-related Kaposi sarcoma histological grade and in vitro behavior: reduced exogenous growth factor requirements for isolates from high grade lesions. AB - Kaposi sarcoma, the most common AIDS-associated malignancy, affects 10-30% of all AIDS patients. To date, research into the biological characteristics of AIDS related Kaposi sarcoma (AIDS-KS) derived cell lines has been based on cultures established from skin explants or pleural effusions/peritoneal fluids. We have established several AIDS-KS lines from biopsy confirmed oral mucosal and epidermal AIDS-KS lesions and have found a correlation between AIDS-KS lesional grade and in vitro cellular growth characteristics. In comparison to epidermal AIDS-KS lesions, mucosal AIDS-KS lesions frequently possessed both a more advanced histologic grade and demonstrated a greater capacity to proliferate in minimal medium. We report the ability of AIDS-KS isolates from high grade lesions to sustain proliferation (greater than 60 population doubling levels) in medium not supplemented with endothelial cell growth supplement and/or cytokine rich conditioned medium. These findings indicate that AIDS-KS cells isolated from high grade lesions have reduced requirements for exogenously provided growth supplements, and suggest that increased autologous cytokine production accompanies AIDS-KS lesional progression. PMID- 7475262 TI - Spontaneous opacification of the lymphatic system during venography. PMID- 7475263 TI - Clustered hepatitis B or human immunodeficiency virus transmission in office settings. PMID- 7475264 TI - On the pathophysiology of arm lymphedema after treatment for breast cancer. PMID- 7475265 TI - A pilot study of the biologic and therapeutic effects of granulocyte colony stimulating factor (filgrastim) in patients with acute myelogenous leukemia. AB - In a carefully monitored pilot study, the in vivo biologic effects of filgrastim were investigated in eight patients with relapsed/refractory acute myelogenous leukemia. Within each patient, filgrastim was administered as a single agent prior to any chemotherapy in escalating doses of 0.12-6.0 micrograms/kg/day as a continuous intravenous infusion. The dose was increased every 14 days until an ANC of > or = 2500/mm3 had been achieved or there was evidence of proliferation of the leukemia. In patients who demonstrated growth of the leukemic clone, cytosine arabinoside was initiated at 200 mg/m2/day for 5 days. Through-out the course of therapy, the effects of filgrastim on maturation and proliferation were assessed by in vitro studies of bone marrow aspirates. Three patients demonstrated a sustained increase in ANC; one achieved a partial remission and remained on therapy for 31 weeks. Two of the three responding patients had hypocellular marrows at the time of initiating filgrastim and demonstrated a low but normal pattern of growth in CFU-GM assay early in the treatment course. This suggested that these two characteristics may define an environment in which filgrastim can induce a growth advantage for the normal residual hematopoietic elements. In this study of selected patients, filgrastim appeared safe. PMID- 7475266 TI - Intensive chemotherapy for poor prognosis myelodysplasia (MDS) and secondary acute myeloid leukemia (sAML) following MDS of more than 6 months duration. A pilot study by the Leukemia Cooperative Group of the European Organisation for Research and Treatment in Cancer (EORTC-LCG). AB - We conducted a prospective, multicenter pilot study of remission induction therapy in patients with poor prognosis MDS and AML evolving from a preceding phase of MDS. Fifty evaluable patients from 15 institutions were treated with one or two remission-induction courses consisting of i.v. idarubicin 12 mg/m2/day on days 1, 2, and 3 combined with a continuous i.v. infusion of cytarabine of 200 mg/m2/day on days 1 to 7. Of the 27 complete remitters (54%), 23 received a consolidation course which was identical to the remission-induction course except for the idarubicin 12 mg/m2 which was given on day 1 only. Fifteen patients received maintenance therapy consisting of six courses of cytarabine 10 mg/m2, s.c. twice daily, for 14 days. Two complete remitters were allografted and five patients received an ABMT. The median survival of all 50 treated patients was 14 months. The median duration of disease-free survival was 11 months with two patients in CR more than 2 years after entering CR. Twenty-four of the 27 remitters have relapsed. Four patients died during remission-induction therapy, but no patient died as a result of persisting hypoplasia. No fatal complications occurred during the consolidation and maintenance courses. Age and stage of disease had no significant impact on CR rate nor on remission duration. The CR rate was significantly (P = 0.03) higher in patients with only normal metaphases compared to patients with cytogenetic abnormalities. The DFS at 2 years was 33 vs 8%, respectively, for patients without or with cytogenetic abnormalities (P = 0.02). This study shows that patients below the age of 60 years with poor risk features are candidates for treatment with combination chemotherapy. A complete remission rate of more than 50% may be expected. Maintaining remission after remission-induction chemotherapy is a difficult issue. Patients not eligible for allogeneic BMT may be treated with intensive post-remission chemotherapy or autologous BMT. PMID- 7475267 TI - Molecular characterization of a chromosome translocation breakpoint t(11;14)(p13;q11) from the cell line KOPT-K1. AB - Recurrent chromosome translocations involving 11p13 and 14q11 are found in 5-10% of cases of T-ALL. The gene involved in the translocation on chromosome 14 is the T cell antigen receptor alpha or delta. The putative oncogene on chromosome 11 is rhombotin 2 (RBTN2)/translocated in T cell gene 2 (ttg-2), a member of the LIM family of proteins. In this paper we characterize a cell line KOPT-K1 that has a t(11;14)(p13;q11). The breakpoint on chromosome 11 involves an Alu-rich region with the break occurring between two Alu sequences on chromosome 11. In addition, approximately 70 bases from the break on chromosome 11 is a tetranucleotide repeat. Whether either of these structures played a role in the translocation is not known. No heptamer or nonamer sequences, implicated in other rearrangements were found near the breakpoint. The breakpoint on chromosome 11 maps more centromeric than previous translocations of this region. Despite this the RBTN2 gene is highly expressed in KOPT-K1. This cell line will be useful for investigating the role of RBTN2 in leukemogenesis and the mechanism by which the translocation alters the expression of RBTN2. PMID- 7475268 TI - Cytogenetic clonality analysis in myelodysplastic syndrome: monosomy 7 can be demonstrated in the myeloid and in the lymphoid lineage. AB - Bone marrow and blood from three patients with myelodysplastic syndrome (MDS) and monosomy 7 were studied for cell lineage involvement of the chromosomal abnormality. Cytogenetic involvement of the myeloid and erythroid cell lineages in MDS with monosomy 7 has been shown before. Lymphoid subpopulations have also been investigated but generally with negative results. A combined technique of May-Grunwald-Giemsa (MGG) for cell cytology and interphase fluorescence in situ hybridization (FISH) using a chromosome 7 specific DNA probe was applied. Further, immunophenotype and genotype of the cells were simultaneously examined with alkaline phosphatase anti-alkaline phosphatase (APAAP) immunostaining and FISH. The monosomy 7 was found in the blasts and in all or in subpopulations of myeloid and erythroid cells. T cells (CD3+, CD5+) did not appear to be involved. B cells (CD19+, CD22+) showed a normal distribution of FISH spots in two patients. In one patient however the loss of a chromosome 7 was found in approximately 70% of the cells positive for B cell markers including CD79a. The results of this study show that in some cases MDS is a disease arising in a progenitor cell with repopulative abilities restricted to myelopoiesis and erythropoiesis. In other cases, the pluripotent progenitor cells in MDS may show the capacities to differentiate into B lineage lymphoid cells, as well as suggesting that in those instances MDS represents a condition of more primitive transformed hematopoietic ancestor cells. PMID- 7475269 TI - Independent clones of trisomy 12 and retinoblastoma gene deletion in Japanese B cell chronic lymphocytic leukemia, detected by fluorescence in situ hybridization. AB - Trisomy 12 and a deletion of chromosome 13 are the most common chromosome abnormalities in patients with B cell chronic lymphocytic leukemia (B-CLL). We determined the frequencies of these abnormalities in Japanese B-CLL patients by FISH in interphase nuclei. Specimens from 42 patients were analyzed using both DNA probes specific to the centromeric region of chromosome 12 and the retinoblastoma (RB) gene. Among 42 patients, eight had trisomy 12 and 12 had the RB gene deletion. We found aberrations of trisomy 12 and the RB gene deletion in a totally different group of patients. This suggested that the trisomy 12 and the RB gene deletion occur in different clones and the presence of which in the same patient may be rare. Furthermore, the frequency of trisomy 12 (19%) found in Japanese B-CLL was lower than that in Western countries (30-35%). On the contrary, the frequency of the RB gene deletion (28.6%) was almost the same as in European B-CLL (30-35%). These results will be helpful in understanding the leukemogenesis of B-CLL. PMID- 7475270 TI - Bax mutations in cell lines derived from hematological malignancies. AB - Many genes are involved in cell cycle control, DNA repair and induction of cell death. Alterations in these genes have been responsible for the development of cancer as well as for resistance to cancer therapy. Recently, an emerging family of bcl2-like genes has been identified that plays a role in the regulation of cell death. Its members are highly conserved in several domains which have been shown to be important for homodimerization or heterodimerization. The ratio between BAX/BCL2 heterodimers and BAX/BAX homodimers appears to be pivotal in deciding the life of death of a cell. We recently detected mutations in evolutionary highly conserved domains of the bax gene in cell lines derived from hematologic malignancies. Similar artificially generated mutations in other bcl2 like family members bcl2, bclxl, or ced9 have been shown to alter their function. This suggests a role for bax mutations in the multi-step pathogenesis of hematological malignancies. PMID- 7475271 TI - Aberrant expression of p21RAS but not p120GAP is a common feature of myelodysplasia. AB - The RAS gene family encodes signal transducing proteins which are involved in the regulation of cell growth and differentiation. Constitutively 'activating' point mutations of RAS have been detected at high frequency in preleukaemia and acute leukaemia, however, the distribution of expression of p21RAS in normal or preleukaemic primary haematopoietic cells has not been studied. We have examined the expression of p21RAS and its negative regulator/downstream effector, p120GAP, in combination with lineage-specific cluster of differentiation markers in normal and preleukaemic myeloid bone marrow cells using flow cytometry. Normal bone marrow was characterized by low and uniform levels of p21RAS expression throughout all lineages analysed. In contrast, 28% (9/32) of patients with myelodysplastic syndrome (MDS) over-expressed p21RAS. In three of these patients a single over-expressing peak of p21RAS expression was observed, with no evidence of a population exhibiting expression within the normal range. In 6/32 MDS patients over-expression of p21RAS was observed only in a subpopulation of the myeloid cells. Follow-up samples were analysed in three of these six patients; over-expression was confirmed in each patient and in two patients a relative expansion of the over-expressing cell population was observed. Eight out of nine of the patients with aberrant p21RAS expression were diagnosed with low-risk MDS. From 21 MDS patients screened for p120GAP expression, no reduction or loss of expression was observed, however, one AML patient demonstrated a heterogeneous pattern of expression. p120GAP expression was lower (P < 0.05) in the AML group than in normals. The results of the study suggest that over-expression of the RAS gene product, p21RAS, may represent an alternative or additional mechanism of activation of the RAS signalling pathway and that this may play a role in leukaemogenesis, however, there is no evidence from this study that loss of p120GAP expression is a feature of myelodysplasia. PMID- 7475272 TI - Expression of bcl-2 mRNA and protein in leukemia-lymphoma cell lines. AB - The expression of the proto-oncogene bcl-2 was examined in a panel of 75 continuous human leukemia-lymphoma cell lines originated from different hematopoietic cell types. The presence of the bcl-2 protein, as evidenced by Western blotting, and its mRNA, as determined by Northern blotting, were not restricted to cells with the chromosomal translocation t(14;18)(q32;q21), but were also detected in a large number of cell lines without t(14;18). The amount of the bcl-2 protein and mRNA in the cell lines with t(14;18) was in the same order of magnitude as in other bcl-2 expressing cell lines of the same lineage, but without the translocation. Bcl-2 was found in all types of hematopoietic cell lines which were assigned to the following lineages based on their phenotypical characteristics: pre-B, B, plasma, T, myeloid, monocytic, erythroid megakaryocytic and Hodgkin's lymphoma derived cell lines. The levels of accumulated mRNA and protein corresponded fairly well in most of the cell lines examined. Our results suggest the notion that bcl-2 expression is widely present in hematopoietic cell lines without restriction to single lineages and, in fact, clearly independent of the chromosomal aberration t(14;18). It is conceivable that bcl-2 expression is a common feature in established hematopoietic cell lines and may contribute to their unlimited growth in vitro. PMID- 7475274 TI - pIXY321 protects against Ara-C or taxol-induced apoptosis and loss of clonogenic survival of normal human bone marrow progenitor cells. AB - By suppressing apoptosis, hemopoietic growth factors (HGFs) promote the survival of CD34+, HLA-DR+ marrow cells that are enriched for hemopoietic progenitor cells (HPC). In the present studies, we have examined the effects of pIXY321, a genetically engineered fusion protein of GM-CSF and IL-3 (GM-CSF/IL-3), on high dose Ara-C (HIDAC) and taxol-induced apoptosis and survival of a multilineage HPC, the CFU-GEMM. Exposure to 1.0 mumol/l taxol for 24 h or HIDAC > or = 10 mumol/l for 4 h induced internucleosomal DNA fragmentation and the morphologic features of apoptosis in CD34+, HLA-DR+ cells. These treatments were associated with > or = 50% inhibition of the assayable CFU-GEMM colony numbers. Incubation in serum-free medium (SFM) alone for 24 h also induced apoptosis of CD34+, HLA DR+ cells, which was associated with reduced intracellular levels of the bcl-2 gene product p26BCL-2. Co-treatment with pIXY321 (10 ng/ml) inhibited apoptosis of CD34+, HLA-DR+ cells incubated in SFM, without significantly increasing the intracellular p26BCL-2 levels. Furthermore, co-treatment with pIXY321 significantly reduced taxol- and Ara-C-induced apoptosis and promoted the survival of CFU-GEMM (P < 0.05). Taxol and Ara-C mediated apoptosis of CD34+, HLA DR+ cells, and its inhibition by pIXY321, was not accompanied by any significant alteration in the intracellular p26BCL-2 levels. By demonstrating that co treatment with pIXY321 confers significant protection against apoptosis of CD34+, HLA-DR+ cells as well as promotes survival of normal HPC exposed to clinically relevant schedules and concentrations of taxol of Ara-C, these results support the design of chemotherapy regimens incorporating pIXY321 plus taxol and/or high dose Ara-C for solid tumors and/or acute leukemias. It is hoped that the use of such a cytokine might maintain normal HPC numbers following chemotherapy, therefore avoiding prolonged suppression. PMID- 7475273 TI - Characterization of clonal immunoglobulin heavy chain and I cell receptor gamma gene rearrangements during progression of childhood acute lymphoblastic leukemia. AB - Instability of antigen receptor gene rearrangements during progression of acute lymphoblastic leukemia (ALL) has important implications for polymerase chain reaction (PCR)-based techniques using these genes for the detection of minimal residual disease (MRD). Antigen receptor gene instability may lead to false negative results in bone marrow samples taken during remission. Utilizing the PCR and consensus primers for rearranged immunoglobulin heavy chain (IgH) and T cell receptor gamma (TCR gamma) gene sequences, we analyzed the bone marrow samples at diagnosis and first relapse for 37 children with ALL. The incidence of clonal evolution at the IgH locus was 9/33 (27%) and at the TCR gamma locus 1/15 (7%). In four of the nine patients with clonal evolution at the IgH locus, the sequence at relapse retained the diversity and joining region (D-N-J) sequences from diagnosis. Patients with clonal evolution were characterized by a higher incidence of more than one IgH PCR band at diagnosis and by late relapse (> 18 months from diagnosis). These results suggest that, where possible, patients with more than one IgH PCR rearrangement at diagnosis should be monitored using another antigen receptor gene, such as TCR gamma, since evolution for this gene was found to be a rare event. By combining this approach with a strategy directed at the more stable D-N-J region of the IgH gene, MRD false negativity would have occurred in less than 10% of patients in the present study. PMID- 7475275 TI - Blast cell proliferative activity and sensitivity to GM-CSF in vitro are associated with early response to TAD-9 induction therapy in acute myeloid leukemia. AB - The current study was undertaken to determine the relevance of leukemic blast cell proliferative activity, cellular parameters of Ara-C metabolism and the in vitro sensitivity to GM-CSF in association with the clinical response to TAD-9 induction therapy in 66 patients with de novo acute myeloid leukemia (AML). Proliferative activity was assessed by 3H-thymidine (3H-TdR) incorporation and thymidine kinase (TK) activity, parameters of Ara-C metabolism comprised the activities of deoxycytidine kinase (DCK) and DNA polymerase alpha (poly alpha) as well as Ara-CTP concentrations and 3H-Ara-C uptake into DNA. GM-CSF sensitivity was determined by in vitro incubation of blasts for 48 h with or without GM-CSF (100 U/ml) followed by an additional 4 h concurrent exposure to GM-CSF and 3H-TdR (0.5 microCi/ml). The following results were obtained as expressed by median values and ranges: 3H-TdR incorporation: 1.07 pmol/10(5) cells (0.0-10.1), TK: 7.3 pmol/min/mg protein (1.3-56.0), DCK: 9.3 pmol/min/mg protein (0.77-47.1), poly alpha: 1.7 pmol/min/mg protein (0.00-28.9), Ara-CTP: 53.3 ng/10(7) cells (13.3-211.0), 3H-Ara-C uptake: 0.06 pmol/10(5) cells (0.0-0.57). 3H-Ara-C uptake was correlated with 3H-TdR incorporation (r = 0.74) and with the (S-phase dependent) activities of TK (r = 0.73) and poly alpha (r = 0.71, but not with DCK activity or intracellular Ara-CTP content. Blast cells of 37 from 55 analyzed patients were found to be sensitive to GM-CSF stimulation as defined by an increase in 3H-TdR incorporation > or = 1.5-fold over control values after the 48 h GM-CSF exposure. In vitro data were related with clinical response to TAD-9 induction therapy in 43 patients with newly diagnosed AML, taking the blast cell reduction at day 10 or 16 to < 5% or > or = 5% residual blasts as early parameter for adequate or inadequate response, respectively. While neither 3H-Ara-C uptake, nor intracellular Ara-CTP concentration, TK nor DCK activity were predictive for response, a high 3H-TdR incorporation and a high poly alpha activity were associated with adequate blast cell reduction. Median values of 3H-TdR incorporation were 2.26 pmol/10(5) cells for patients with adequate blast cell clearance and 0.80 pmol/10(5) cells for patients with inadequate blast cell clearance (P = 0.11), the respective values for poly alpha were 3.22 pmol/min/mg protein for responders and 1.1 pmol/min/mg protein for non-responders (P = 0.0085).(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 7475277 TI - Expression of the multidrug resistance P glycoprotein in newly diagnosed adult acute lymphoblastic leukemia: absence of correlation with response to treatment. AB - We analyzed P glycoprotein (PGP) expression and its correlation with hematological parameters and outcome in 50 cases of newly diagnosed adult acute lymphoblastic leukemia (ALL). PGP expression was evaluated by flow cytometry using MRK16 monoclonal antibody (MoAb) and/or immunocytochemistry on marrow slides, using JSB1 MoAb. Thirty-two of the 50 patients (64%) were PGP positive by at least one of the two methods, which gave concordant results in 15 of the 18 cases in which they were both used. No correlation between PGP expression and clinical and hematological parameters including WBC counts, immunophenotype and karyotype was seen, although there was a trend for more frequent CD34 expression in PGP-positive cases. All patients were treated with intensive chemotherapy. We found no difference in complete remission (CR) rate, actuarial disease-free survival and survival in PGP-positive and PGP-negative cases. Our findings suggest that the clinical significance of PGP expression is less clear in ALL than in AML. Wider use of functional techniques of evaluation of mdr1 gene expression, which assess the 'pumping' activity of PGP, and their correlation with quantitative analysis of mdr1 mRNA and protein, would probably improve knowledge of the role of PGP in ALL. Analysis of other mechanisms of drug resistance, especially multidrug resistance-associated protein (MRP) expression, would also be useful. PMID- 7475276 TI - In vitro chemosensitivity assessed with the MTT assay in childhood acute non lymphoblastic leukemia. AB - Cellular drug resistance is supposed to play a major role in chemotherapy failures which frequently occur in childhood acute non-lymphoblastic leukemia (ANLL). Therefore, we determined in vitro chemosensitivity to daunorubicin, doxorubicin, mitoxantrone, 6-thioguanine, etoposide, and cytosine arabinoside (Ara-C) in childhood ANLL using the colorimetric MTT assay. The 4-day MTT assay was successfully performed in 62/73 samples obtained from 53 children with ANLL. We obtained comparable results from bone marrow or peripheral blood samples, and from fresh or cryopreserved samples. In vitro chemosensitivity was not related to clinical features such as sex, age, white blood cell count, or FAB-types. The group of poor responders to chemotherapy was median 3-fold more resistant to Ara C than the group of good responders, but identification of a threshold for Ara-C sensitivity predictive for individual responses was limited due to the great overlap of in vitro chemosensitivities between both groups. Children with relapsed ANLL were in vitro median 3-fold more resistant to Ara-C than the initial ANLL group. No significant differences for the other drugs were observed with respect to clinical response or disease status. These results suggest that in vitro resistance to Ara-C plays an important role in chemotherapy failures in childhood ANLL, but larger studies are necessary to establish the predictive value of Ara-C sensitivity assessed with the MTT assay. PMID- 7475278 TI - Comparison of antitumor activities of 2-chlorodeoxyadenosine and 9-beta arabinosyl-2-fluoroadenine in chronic lymphocytic leukemia and marrow cells in vitro. AB - The in vitro antitumor activities of the nucleoside analogs, 2 chlorodeoxyadenosine (CdA) and 9-beta-arabinosyl-2-fluoroadenine monophosphate (Flu), and the alkylating agent, chlorambucil (CLB), were compared in leukemic cells from 28 patients with chronic lymphocytic leukemia (CLL). On a molar basis, the median sensitivities of the cells to these agents were CLB > CdA > Flu. CLL cells from 90% of the patients had similar relative orders of sensitivities to CdA and Flu, while cells from 10% of the patients showed differential sensitivities to these agents. There was no relationship between the sensitivities of the cells to the nucleoside analogs and sensitivity to CLB. CdA and CLB produced similar toxicities to human marrow progenitor cells in vitro, while Flu was less toxic to these cells. An 18 h exposure to CdA produced significantly greater cell kill of both CLL and marrow progenitor cells than an equivalent 2 h treatment; however, the difference in cytotoxicity was greater for the tumor cells resulting in a higher therapeutic index with the 18 h treatment. The intracellular accumulation of drug varied 5-fold for CdA, with the major metabolite being CdAMP, and 15-fold for Flu, with the major metabolite being F ara-ATP. However, the accumulation of CdA, Flu or their metabolites did not predict for drug sensitivity. These studies suggest that CdA and Flu cross resistance cannot be assumed in all CLL patients. The therapeutic effectiveness of CdA may be enhanced by use of a prolonged, low-dose drug regimen. PMID- 7475279 TI - Discordant P-glycoprotein antigen expression and transport function in acute myeloid leukemia. AB - Expression of the multidrug resistance efflux pump P-glycoprotein (Pgp) was measured in a series of AML patients using two flow cytometry methods. Transport function was assessed by measuring the modulating effect of the Pgp inhibitor cyclosporin A (CsA) on the cellular accumulation of daunorubicin, and Pgp antigen expression by surface immunofluorescence using the MRK-16 antibody. Both methods showed a wide range of values for Pgp expression between individual patients, but in contrast to a series of cell lines expressing Pgp there was no correlation between antigen expression and transport function in the clinical samples. As previously reported for chronic lymphocytic leukemia (CLL), pretreatment with neuraminidase markedly improved MRK-16 staining in some cases, indicating that abnormal glycosylation can cause epitope masking in AML blasts. Because experience with cell lines shows that Pgp expression is a continuous variable which correlates with the level of drug resistance, rather than the 'positive' or 'negative' which are frequently reported by clinical flow cytometry laboratories, we used a calibration procedure to estimate the actual number of Pgp molecules expressed in the AML samples. Despite the additional refinements of neuraminidase treatment and antigen quantification, the correlation between Pgp antigen expression and daunorubicin accumulation remained extremely weak (r = 0.11; P = 0.63). It is suggested that the assay for transport function can detect molecules that affect daunorubicin accumulation but are antigenically distinct from classical P-glycoprotein. Heterogeneity of multidrug resistance efflux pumps might in part explain the relatively weak prognostic significance of immunofluorescence detection of Pgp in AML patients. PMID- 7475280 TI - Triazene compounds induce apoptosis in O6-alkylguanine-DNA alkyltransferase deficient leukemia cell lines. AB - Previous studies demonstrated that triazene compounds (TZC) possess antitumor, antimetastatic and immunosuppressive activity, and induce novel antigenic properties in neoplastic cells. Moreover, TZC showed marked antitumor activity in patients with acute myelogenous leukemias (AML). In most cases leukemic blasts with low levels of the repair enzyme O6-alkyl-guanine-DNA alkyltransferase (OGAT) were highly susceptible to TZC. Therefore the cytotoxic effects of TZC against human leukemic cells and the influence of OGAT modulation were investigated. Five leukemia cell lines were treated with the in vitro active derivative of dacarbazine: 5-(3-methyl-1-triazeno) imidazole-4-carboxamide (MTIC), or with temozolomide (TZM), which is readily cleaved to form the linear triazene MTIC in aqueous solution. The results showed that treatment with TZC at concentrations ranging between 62.5 and 250 microM significantly inhibited cell growth of U-937 and K-562 leukemia cell lines, both with undetectable OGAT activity. Growth inhibition was accompanied by DNA fragmentation and reduction of cell volume characteristic of cell undergoing apoptosis. In contrast, Daudi, HL-60 and Jurkat leukemia cell lines, characterized by high levels of the repair enzyme, were resistant to concentrations of TZC up to 500 microM. Treatment of resistant lines with O6-benzylguanine (BG, a specific inhibitor of OGAT) rendered HL-60 and Daudi but not Jurkat cells sensitive to cytotoxic effects and apoptosis mediated by MTIC. The results presented suggest that: (1) apoptosis is involved in cytotoxic activity of TZC; (2) OGAT could have a role in preventing programmed cell death induced by TZC; and (3) treatment with BG could potentiate cytotoxic and apoptotic effects of TZC on leukemic cell lines when high level of OGAT activity is the main factor involved in resistance to TZC. PMID- 7475281 TI - Phenotypic analysis of CD34 subpopulations in normal human bone marrow and its application for the detection of minimal residual disease. AB - In the past, studies on CD34+ cells have been based on the use of monoclonal antibodies conjugated with different fluorochromes that show different fluorescence intensity and yield variable results. Moreover, most of these studies have neither specifically focused on adult human BM samples nor have they used combinations to explore specifically the phenotype of myeloid committed CD34+ cells. The aim of the present study has been to characterize the normal human CD34+ precursor cells from adult BM in order to identify missing or extremely rare phenotypes that can be used for detecting minimal residual disease (MRD) in patients with AML. For this purpose we have utilized the fluorochrome conjugates that provide the most sensitive signals for identifying low antigenic expression, and the technique has been adapted to the characterization of cells present at very low frequencies. Normal human BM samples from 13 adult healthy volunteers have been analyzed using triple stainings at flow cytometry. The mean percentage of CD34+ cells detected was 0.72 +/- 0.33%; these cells displayed an heterogeneous light-scatter distribution. Most CD34+ cells coexpressed CD38 (96.7 +/- 5.7%), HLADR (81.6 +/- 14.0%), CD33 (84.7 +/- 18.3%), CD13 (84.6 +/- 16.2%) and CD71 antigens (65.5 +/- 9.1%). In addition, almost half of CD34+ cells were CD117+ (60 +/- 26.8%). Only a small proportion of CD34+ cells coexpressed CD4 (15.5 +/- 11.7%, CD36 (31.7 +/- 6.2%), CD61 (16.3 +/- 12.9%), CD41 (6.5 +/- 5.5%) or the lymphoid associated markers CD10 (18.6 +/- 11.8%) and CD19 (12.3 +/- 13.2%). Reactivity for the CD15 antigen was observed in a small population of CD34+HLADR+ cells (11.6 +/- 11.2%) although its intensity of expression was lower than that of the more mature granulocytic cells. No CD34+ cells displayed CD14, CD65, CD20, strong CD22, CD3 and CD56 antigens. Accordingly, most adult bone marrow CD34+ cells appeared to be committed to the myeloid lineage (CD13+/CD33+) and displayed an intermediate-to-large FSC/SSC while the lymphoid-committed CD34+ cells (CD19+, CD10+) were in a minority with low FSC/SSC values. By triple marker stainings several phenotypes of CD34+ precursor cells were found to be either undetectable or present at very low frequencies (< 1 x 10(-3)) in the normal human adult bone marrow. These data may be of great value for defining leukemia 'associated' phenotypes used to detect minimal residual disease in adult acute leukemia patients. PMID- 7475282 TI - Improved vaccination response during ranitidine treatment, and increased plasma histamine concentrations, in patients with B cell chronic lymphocytic leukemia. AB - Patients with B cell chronic lymphocytic leukemia (B-CLL) have decreased capacity to mount relevant antibody responses upon immunization, and development of hypogammaglobulinemia is part of the natural history of the disease. We investigated the influence of histamine type-2 (H2) receptor blockade by ranitidine on the in vivo antibody production in B-CLL patients following vaccination. Anti-polysaccharide antibodies in B-CLL patients, vaccinated with a tetanus-toxoid conjugated vaccine against Haemophilus influenzae type-B (Hib), reached long-term protective levels in more than 90% of B-CLL patients randomized to ranitidine treatment, as compared to 43% of the untreated patients (P = 0.024). No difference in the response to vaccination against influenza virus types A and B protein could be detected between the two groups. Plasma histamine levels were 2-fold to 20-fold higher in 23 out of 31 B-CLL patients, compared to normal controls, and these levels showed a significant positive correlation to disease duration. These findings indicate the possibility of improving in vivo antibody production against a highly relevant pathogen in B-CLL patients by histamine type-2 receptor blockade, and the combined finding of an immune stimulatory effect of ranitidine and increased plasma histamine levels, strongly suggests the involvement of histamine in the pathogenesis of B-CLL immunodeficiency. PMID- 7475283 TI - Effects of interleukin 10 on blast cells derived from patients with acute myelogenous leukemia. AB - The effect of interleukin 10 (IL-10) on proliferation and cytokine secretion by acute myelogenous leukemia (AML) blast cells was investigated in vitro. IL-10 inhibited spontaneous AML blast proliferation for a majority of patients, whereas in the presence of exogenous growth factors (granulocyte-stimulating factor, G CSF; granulocyte-macrophage colony-stimulating factor, GM-CSF; interleukin 3) the IL-10 effect on blast proliferation showed a wide variation depending on the individual AML patient. IL-10 seemed to cause an irreversible inhibitory effect on AML blasts, as inhibition could also be demonstrated when IL-10 was present only during the initial preincubation of the leukemia cells. IL-10 also inhibited AML blast colony formation. However, independent of the effect on AML blast proliferation, IL-10 decrease cytokine secretion from AML blast cells for all patients, as demonstrated for IL-1 alpha, IL-1 beta, tumor necrosis factor-alpha, GM-CSF and interleukin 6. IL-10 did not inhibit development of apoptosis in AML blasts cultured in vitro. Expression of complement receptors and capability to adhere and internalize bacteria by AML blasts were not altered by IL-10. PMID- 7475284 TI - Cell cycle kinetics in childhood acute leukemia studied with in vitro bromodeoxyuridine labeling, Ki67-reactivity, and flow cytometry. AB - Cell cycle kinetics of childhood acute leukemia were determined by the in vitro labeling of marrow blast cells with bromodeoxyuridine (BrdUrd) and subsequent flow cytometry of BrdUrd/DNA and Ki67/DNA in 18 patients with acute lymphocytic leukemia (ALL) and eight patients with acute nonlymphocytic leukemia (ANLL). The BrdUrd-labeling index (BrdUrd-LI) and the duration of S phase (Ts) were calculated from the slope of the regression line obtained by plotting the serial labeling indices against the labeling time. The Ts and potential doubling time (DTpot) of marrow leukemia cells varied from 6.1 to 34.3 h (median 14.3 h) and 1.1 to 20.7 days (median, 7.3 days), respectively. The duration of the total cell cycle time (Tc) which was determined by the Ki-67-derived growth fraction (Ki-67 GF) varied from 14.0 to 112.5 h (median 43.2 h). BrdUrd-LI, DTpot, Ki-67-GF and Tc were significantly correlated with the subtypes (early B-ALL, T/B- ALL and ANLL) of the disease. The median values of LI and GF were much lower in ANLL than in ALL. However, the low proliferative activity of ANLL was not accompanied by a prolonged duration of the total cell cycle time. The longest median duration of Tc was noted in early B-ALL (75.2 h) and the median Tc in ANLL (36.7 h) was close to that in T/B-ALL (34 h). Ts appeared to be rather independent of subtypes of the disease. These results show that there are distinct in vitro growth characteristics in relation to the subtypes of childhood acute leukemia. PMID- 7475286 TI - Frequent expression of shared idiotypes in mantle cell lymphoma and extranodal small lymphocytic/non-mantle cell diffuse small cleaved lymphoma. AB - A panel of 14 anti-shared idiotype (Sid) monoclonal antibodies selected according to their high cross-reactivity to various lymphomas was immunohistologically tested for reactivity with seven reactive lymphoid tissue specimens and 227 B cell lymphoma specimens obtained from Japanese patients. In the reactive lymphoid tissues, the anti-Sld antibodies each reacted with a subpopulation of cells in the mantle zone and interfollicular areas; they rarely reacted with cells in the germinal center. In the B cell lymphomas, 13 anti-Sld antibodies reacted with a total of 78 of 186 (42%) specimens bearing immunoglobulin; none of the antibodies reacted with 41 specimens not bearing immunoglobulin. In mantle cell lymphomas (15/19, 79%) and extranodal small lymphocytic/non-mantle cell diffuse small cleaved lymphomas (11/15, 73%), the reactivity of the antibodies was high compared with that in the other lymphomas (52/152, 34%; P = 0.0002 and 0.004, respectively), including follicular lymphomas (11/42, 27%; P = 0.002 and 0.002, respectively). Since idiotypes are associated with the hypervariable regions and antigen-binding sites of immunoglobulin, these findings may reflect the differences in the regions/sites in each of these diseases. PMID- 7475285 TI - Establishment and characterization of a novel cell line, TK-6, derived from T cell blast crisis of chronic myelogenous leukemia, with the secretion of parathyroid hormone-related protein. AB - We established a novel T cell line, designated TK-6, from a patient with T cell lineage blast crisis of chronic myelogenous leukemia (CML) complicated by hypercalcemia. A surface marker study showed T cell phenotype, cluster designation (CD)4, CD5 and CD7. Light and electron microscopic examination revealed myeloperoxidase (MPO)-negative, however, ultrastructural examination under certain specific conditions demonstrated that some cells were MPO-positive. The TK-6 cell karyotype carried a t(9;22)(q34;q11) and additional chromosome aberrations, including a deletion of the long arm of chromosome 6 and the abnormality of chromosome 7. Southern blot analysis showed rearrangement of the T cell receptor beta-chain (TCR beta) gene and the major breakpoint cluster region (bcr) gene. Northern blot analysis detected the expression of the parathyroid hormone-related protein (PTHrP) gene, however, the proviral genome of human T cell leukemia virus type I (HTLV-I) was negative. This cell line will provide a valuable resource for the analysis of the relationship between T cell lineage crisis and myeloid differentiation and for the analysis of humoral hypercalcemia of malignancy (HHM) or leukemia. PMID- 7475287 TI - The effects of passive anti-viral immunotherapy in AKR mice: II. Susceptibility to B cell lymphomagenesis. AB - Prevention of high frequency spontaneous T cell lymphoma development in AKR mice by mAb 18-5 treatment was shown to involve inhibition of the recombinant Class I MCF virus formation and elimination of the early occurring potential lymphoma cells (PLCs). A low B cell lymphoma incidence (16% at a mean latency of 540 days) and a low level of PLCs (yielding 12% B cell lymphoma development following lymphoid cell transfer) was observed in mAb 18-5 treated mice (in contrast to a high PLC level in thymectomized AKR mice that could be experimentally triggered to progress to overt CD5+ B cell lymphomas). Administration of anti CD8 mAb or IL 4 to 12-month-old mAb 18-5 pre-treated mice only slightly increased B cell lymphoma incidence (up to 30-40%). Exposure to split-dose irradiation resulted in 26% B cell lymphomas at a 250 day mean latency. The phenotypes of the B lymphomas developing in mAb 18-5 treated mice were: B220+ (14.8+, 6B2+), 6C3+, Mac2+, CD5-. Most lymphomas expressed l-a and surface IgM, pointing to their mature B cell characteristics. Moreover, in some of the lymphomas, high levels of IgM production and secretion were determined. A comparison of the morphological characteristics (based on light and ultrastructure microscopy) of CD5+ and CD5- B cell lymphomas developing in AKR mice indicated marked differences. Analysis of the IgH locus of representative CD5- B lymphomas showed an identical pattern of IgH rearrangement in some tumors (similar to previous findings among CD5+ lymphomas). The virological analysis of the CD5- B cell lymphomas (similar to those observed in the CD5+ B cell lymphomas of AKR origin) showed that their development did not require formation of the pathogenic MCF recombinant viruses. The differences observed between the CD5+ and CD5- B cell lymphomas developing in AKR mice (following prevention of spontaneous T cell lymphomagenesis) may be due to their origin of different B cell precursors or from B cells at different levels of differentiation. PMID- 7475288 TI - A CD10-positive subset of malignant cells is identified in multiple myeloma using PCR with patient-specific immunoglobulin gene primers. AB - Immunophenotypic studies show the presence of CD10-bearing malignant cells in a small subset of multiple myeloma (MM) patients. We used a sensitive PCR-based technique in order to determine the frequency that MM patients contain a malignant subpopulation which expresses this antigen. The immunoglobulin (Ig) heavy chain variable region (VH) gene sequence expressed by the malignant clone in MM can be used as a tumor specific marker. After determining this sequence in six MM patients, patient specific VH oligonucleotide primers from complementarity determining region (CDR) sequences were generated. Bone marrow mononuclear cells from these patients were incubated with two different anti-CD10 antibodies or isotype identical murine IgG controls. Cells were then sorted by flow cytometry into the 1% brightest cells containing > 99.99% CD10-positive cells and two fractions including the 90 and 10% dimmest staining cells. PCR amplification was performed on DNA from approximately 10(4) cells (0.1 microgram) using patient specific CDR1 and CDR3 primers. Detectable PCR product was obtained in each sorted sample although the intensity of the band was much higher in cells lacking CD10 expression (the 90 and 10% dimmest fractions) than in the CD10-bearing (1% brightest) population. These results imply that there is a small population of CD10-bearing clonal cells in most, if not all patients with MM. PMID- 7475289 TI - Monitoring of leukemia growth in a rat model using a highly sensitive assay for the detection of LacZ marked leukemic cells. AB - A very sensitive assay for the detection of LacZ marked cells of an in vitro growing subline of the brown Norway rat myelocytic leukemia (BNML) model was developed. By combining cytochemical X-gal staining with D-galactose mediated suppression of endogenous background beta-galactose activity, a detection sensitivity of one leukemic cell per 10(8) normal bone marrow cells could be achieved. A detailed analysis of the in vivo growth pattern and kinetics of this cell line is presented. Also, it is shown that after cyclophosphamide treatment of leukemic rats no leukemic colonies are formed in an agar-colony assay, whereas the leukemic cells remain detectable in the bone marrow for a considerable time period. Eventually, however, all leukemic cells disappear from the marrow. These findings are discussed in the light of prolonged detection of rare leukemic cells in patients in continuing remission. PMID- 7475290 TI - Evaluation of in vivo induction of apoptosis in patients with acute leukemia treated on a phase I study of paclitaxel. PMID- 7475291 TI - Absence of hereditary p53 mutations in eight familial Hodgkin's disease pedigrees. PMID- 7475292 TI - Granulocytic sarcoma in infant with MLL rearrangement preceding acute monoblastic leukemia with t(10;11)(p11;q23). PMID- 7475293 TI - Transformation of chronic lymphocytic leukemia to multiple myeloma: clonal evolution of second malignancy? PMID- 7475294 TI - Familial occurrence of cutaneous T cell lymphoma: a case report of monozygotic twin sisters. PMID- 7475295 TI - Platelet-activating factor concentrations in bone marrow of patients with lymphoid and nonlymphoid hematologic malignancies. PMID- 7475296 TI - Gene Therapy: New Frontiers. Symposium proceedings. Dublin, Ireland, September 18 21, 1995. PMID- 7475297 TI - 1994 de Villers Lecture--tribute to Denis Burkitt. PMID- 7475298 TI - Lymphosarcoma in horses. PMID- 7475299 TI - Leukemia cell derived factor (LDF) induces hematopoiesis. AB - A leukemic cell line, A1, derived from a child with acute lymphoblastic leukemia was observed to have early B-cell lineage. When irradiated A1 cells were cultured with normal bone marrow cells in presence of recombinant erythropoietin (rhEPO), it stimulated burst forming unit erythroid (BFU-E). In order to determine if other leukemia cells or cell lines also induce similar stimulation, we used different leukemic cell lines and fresh B-cells from patients with chronic lymphocytic leukemia in bone marrow bioassay. Our results suggest that not all leukemic cells induce BFU-Es. To further identify the leukemia cell derived factor (LDF), A1 cells were lysed and differentially centrifuged to separate plasma membrane protein from cytosolic and nuclear proteins. Only membrane associated protein were shown to induce BFU-E. However, when spent medium was collected and used in the bioassay, it also stimulated BFU-E, suggesting LDFs may be shed from plasma membrane. To further identify LDF, A1 plasma membrane associated proteins were separated by ion-exchange FPLC. Protein fractions were collected and were used in the bioassay to identify the fraction that induced BFU Es. PMID- 7475300 TI - The effect of epidermal growth factor receptor (EGFR) expression on in vivo growth of rat C6 glioma cells. AB - The discovery of EGFR gene amplification in glioblastoma multiforme has prompted interest in experimental therapies to target the receptor on brain tumor cells. To develop an animal model for in vivo study of such strategies, we transfected C6 glioma cells with a plasmid containing the neomycin resistance gene and the human EGFR gene under the control of the glucocorticoid-inducible MMTV promoter. Following selection with G418, individual clones that expressed EGFR at high levels were selected. Kinetics of EGF binding fit a dual site model indicating the presence of both high (KA = 2.5 x 10(9) M-1) and low (KA = 3.3 x 10(7) M-1) affinity receptors. To assess growth in vivo, graded numbers of either wild-type or transfected cells were implanted into the brains of CD Fischer 344 rats. No differences in survival were observed between groups of animals injected with either wild-type or transfected cells at inocula of 10(3) or 10(4) respectively. In addition, one-third of animals (7/21) challenged with 10(5) or 10(6) transfected cells survived > 50 days compared to 0% of animals (0/12) challenged with 10(5) or 10(6) wild-type cells. Such an effect suggests greater immunogenicity of transfected cells, but only at the larger inocula. Since C6 glioma cells will grow in both outbred and inbred strains, our model should have a number of applications including the in vivo study of EGFR targeting for glioma therapy. PMID- 7475302 TI - Apoptin, a protein encoded by chicken anemia virus, induces cell death in various human hematologic malignant cells in vitro. AB - Apoptin, a small protein encoded by chicken anemia virus (CAV) was expressed in various human hematologic malignant cell lines derived from leukemias and lymphoma. Three of these cell lines contain bcl-2 or BCR-ABL proteins, known to block apoptosis induced by chemotherapeutic compounds. By immunofluorescence and propidium-iodide staining apoptin was shown to induce apoptosis in all analysed cell lines. Early after expression, apoptin exhibited a fine-granular distribution in the still intact nucleus. Later, apoptin became aggregated and the nucleus segmented. The data with truncated apoptin indicate that for optimal induction of apoptosis apoptin has to be located in the nucleus. PMID- 7475301 TI - Serum G-CSF levels in patients undergoing G-CSF/chemotherapy mobilized peripheral stem cell harvest and predictors of neutrophil and platelet recovery. AB - Granulocyte colony-stimulating factor (G-CSF)/Chemotherapy mobilized peripheral blood progenitors are an effective source of stem cells which affords rapid and complete hematopoietic engraftment after myeloablative chemotherapy regimens. The dose of G-CSF most commonly used for mobilization is 5 micrograms/kg. We measured G-CSF levels in patients with chemosensitive malignancies undergoing peripheral stem cell harvest to determine whether there was a relationship between serum G CSF levels and the yield of CD34+ hematopoietic progenitors. Peripheral blood stem cells (PBSCs) were mobilized by chemotherapy followed by G-CSF (5 micrograms/kg) started 24 hours after completion of chemotherapy. PBSCs were collected by daily leukapheresis during G-CSF stimulation once the WBC had recovered to 1.0 x 10(9)/L, with 10 liters of blood processed using a Fenwall CS 3000. G-CSF levels were monitored daily before and after leukapheresis. CD34+ cells from daily leukapheresis collections were determined in 11 patients. Immunophenotyping analyses of CD34+ and non-CD34+ cells for surface antigens CD38+, HLA-DR, CD71+, CD61+ and CD42a+ were performed on these daily leukapheresis. The mean (SD) number of days to neutrophil recovery (NR: > or = 0.5 x 10(9)/L) after stem cell reinfusion was 9.2 (1.92). The corresponding values for platelet recovery (PR: > or = 20 x 10(9) L) were 8.1 (2.39) days. Using multiple regression analyses, the best predictors for NR were: last G-CSF (R2 = 0.21); last G-CSF and CD34+ (R2 = 0.67); last G-CSF, CD34+ and number of chemotherapy cycles (R2 = 0.72).(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7475303 TI - Cryopreserved cord blood myeloid progenitor cells can serve as targets for retroviral-mediated gene transduction and gene-transduced progenitors can be cryopreserved and recovered. AB - To determine future possibilities for gene transfer, we evaluated whether myeloid progenitor cells from human umbilical cord blood (CB) could be frozen, thawed in viable form and transduced with a Neomycin resistance (NeoR) gene using retroviral vectors, and if fresh progenitor cells transduced with a Neo gene could be cryopreserved and recovered. Fresh and thawed cryopreserved nonadherent low-density T-lymphocyte depleted (NALT-) CB cells were assayed before and after gene transduction for colony formation in the presence of multiple growth factors in the absence and presence of G418. The results demonstrate that the NeoR gene could be introduced into thawed cryopreserved myeloid progenitor cells at an efficiency similar to that of fresh cells and that fresh cells transduced with the NeoR gene could be frozen in a cryopreserved state and recovered after thawing. Proviral integration, as assessed by PCR/Southern Analysis, confirmed the G418R colony data. Proviral integration was detected not only in primary G418R-colonies, but also in replated colonies in secondary dishes derived from G418R-multipotential progenitor cells (CFU-GEMM) suggesting stable integration of the transduced gene into early subsets of replatable progenitors. This information may be of use clinically. PMID- 7475304 TI - Immunogenization of a murine T-cell lymphoma via transfection with interferon gamma. AB - Tumor cell variants were derived from the BW5147 T-cell lymphoma that differ in major histocompatibility complex (MHC) class I antigen expression, tumorigenicity and metastatic potential. In general, increased H-2Kk expression was found to be correlated with a reduced tumorigenicity and spontaneous metastasis. CD8+ T cells were identified in the immune recognition of such variants, implicating a role for H-2Kk in the presentation of tumor-associated antigens. In the present study, H-2Kk+ BW variants were transfected with a gene encoding interferon-gamma (IFN gamma), a potent inducer of MHC class I expression. The resulting transfectants exhibited an increased expression of H-2Kk and concomitantly an inability to generate visible tumors and a reduced metastatic capacity. Furthermore, immunization with the IFN-gamma transfectants resulted in an increased generation of cytotoxic T lymphocytes (CTLs) that lysed both the transfectants and the parental tumor cells. Based on these results, vaccinations with the IFN-gamma transfectants were performed against the parental tumor cells. The results clearly demonstrated that such vaccinations reduced significantly the tumorigenicity and metastatic capacity of the parental tumor cells. Hence, in this tumor model, IFN-gamma gene transfection provides a means to immunogenize H 2Kk+ BW tumor cells. PMID- 7475305 TI - Manipulation of the immune response by foreign gene expression in the thymus. AB - Retroviral gene transfer vectors have been developed for optimal in vivo gene therapy. Ideally, these vectors should target gene expression specifically to selected tissues or organs. Our studies focus on the development of retroviral vectors for gene delivery to the thymus. The goal of these studies is to utilize thymic expression of exogenous genes to manipulate the immune repertoire. We have characterized the selective thymic tropism of a molecular clone of Gross murine leukemia virus, GD-17, to thymic medullary epithelial cells using immunohistochemical staining and confocal microscopy. Specific expression of viral antigens in the thymus lead to the induction of immunologic tolerance to GMuLV proteins. This tissue specific vector may thus be used to study the requirements of epithelial mediated tolerance induction, and provide a more efficient tool for gene therapy. PMID- 7475306 TI - Glycosylation-dependent peptide antigenic determinants of env gp46 HTLV-1. AB - Human T-lymphotropic virus type 1 (HTLV-I) is a retrovirus associated with adult T-cell leukemia/lymphoma, and the virus infection constitutes a growing public health problem. In a continuing effort to engineer conformationally dependent HTLV-I epitopes that elicit a protective immune response, we have examined the role and functional importance of carbohydrate moieties in specific immune recognition and antibody responses. There have been several reports of the importance of N-linked virus glycosylation in the formation of neutralizing antibodies. Residues 230-257 is predicted to encode two beta-turn/loop regions at 240-244 (LYGPN), 248-257 (VPSSSSTPL) and a glycosylation site at N-244 (NVS). We have successfully engineered and synthesized the 233-253 sequence of gp46 of HTLV 1 with and without GlcNAC at Asn244. Circular dichroism spectroscopy and proton NMR showed the presence of beta-turn conformation in both peptide constructs. Chimeras of the glycosylated and non-glycosylated epitope with promiscuous T-cell epitope were synthesized and shown to elicit high titered antibodies in rabbits specific for the immunogen (SC1MVF and SC2MVF) and the B cell epitope 233-253. Additionally, antibodies to the glycosylated form of the peptide recognized the HTLV-I envelope precursor in radioimmunoassay precipitation assay and react with HTLV-I whole virus preparations in ELISA. PMID- 7475307 TI - Cytokine-mobilized peripheral blood CD34+Thy-1+Lin- human hematopoietic stem cells as target cells for transplantation-based gene therapy. AB - Gene-therapy of blood-borne disorders may be best achieved using hematopoietic stem cells (HSC) which have extensive self renewal potential as well as multilineage repopulating potential as a cellular target. The human HSC, which is CD34+Thy-1+Lin- has been isolated from fetal, adult bone marrow and cytokine mobilized peripheral blood (MPB) (1-3). Results presented in this study show that the degree of mobilization of HSC into peripheral blood of cancer patients is highly variable and that the combined use of high dose chemotherapy and GM-CSF as a mobilization strategy is superior to the use of G-CSF with regard to the mobilization of true HSC. A multistep cell isolation procedure has been developed which utilizes high speed flow-cytometric cell sorting and allows the isolation of sufficient numbers of HSC from MPB to permit their use as an hematopoietic graft for clinical transplantation. Hematopoietic stem cells isolated from MPB are capable of self-renewal and differentiation into multiple hematopoietic lineages as shown by their behavior in both in vitro and in vivo assays. Mobilized PB mononuclear cells isolated from cancer patients are frequently contaminated with tumor cells. Using this cell isolation procedure, HSC preparations from patients with multiple myeloma have been created with greatly reduced tumor cell burdens. These CD34+Thy-1+Lin- cells are capable of being stably transduced at high efficiency (32-75%) by co-culture on a cell line producing recombinant retroviruses containing the neomycin-resistant gene. These HSC cell populations are likely ideal targets for hematopoietic cell-based gene therapy.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7475308 TI - Expression of AML1 and ETO Transcripts in hematopoietic cells. AB - Recently, two genes, AML1 and ETO have been isolated from the chromosomal breakpoint of t(8;21). In this study, we isolated and identified fusion transcripts from a leukemic cell line carrying t(8;21). We demonstrated by PCR analysis that these transcripts are consistently expressed in fresh leukemic cells with t(8;21). On the other hand, the wild type of ETO is expressed in several hematopoietic cells from different lineage, while the expression of AML1 was present in all hematopoietic cells investigated. These widespread expression suggests these molecules play an essential role in hematopoiesis. PMID- 7475309 TI - Selection of a highly enriched population of retrovirus-infected human hematopoietic progenitor cells using SNL fibroblasts. AB - Retrovirus-mediated gene transfer into human hematopoietic progenitor cells for therapeutic or experimental purposes has proved difficult due to low and variable infection efficiency. To address this, we have developed an in vitro system for the selection and maintenance of a highly-enriched population of retrovirus infected hematopoietic progenitor cells. Human umbilical cord CD34+ cells were cultured on SNL, a neo-containing murine fibroblast cell line used for embryonic stem cell culture. SNL-supported CD34+ cultures could be maintained with continuing blast cell and CFU-GM production for eight weeks, compared to four weeks in the absence of SNL. We then tested the ability of SNL to facilitate the selection in G418 of CD34+ cord cells infected with the neo-containing retrovirus, vsn-2. While all cells in the control cultures died within 14 days, vsn-2-infected CD34+ cells continued to proliferate, differentiate and produce CFU-GM for up to five weeks after infection. 100% of individually-plucked CFU-GM from such cultures were shown by PCR to be successfully infected. This approach should be useful for experimental work and, since it would diminish competitive repopulation between infected and uninfected progenitors, may also be utilized, with modification, for optimizing gene therapy protocols. PMID- 7475310 TI - Increased resistance to methotrexate in human hematopoietic cells after gene transfer of the Ser31 DHFR mutant. AB - Retroviral gene transfer into hematopoietic cells has many experimental as well as clinical applications. Although transduction efficiency of retroviral vectors is higher than with conventional methods, selection of successfully transduced cells may become mandatory for efficient in vivo transfer. We have been evaluating a retroviral construct that meets the criteria for a clinically acceptable selection system. The Ser31 dihydrofolate reductase (DHFR) mutant confers resistance to methotrexate (MTX) due to decreased binding of the drug. while its enzymatic activity remains adequate for normal folate metabolism. Transduction of this vector into murine hematopoietic cells has been recently described and increase in MTX resistance could be observed. We investigated transduction of CD34-antigen positive subpopulations of hematopoietic progenitor cells and CD34-positive/CD38-negative subpopulations enriched for stem cells. We focused on two sources of primary hematopoietic cells, umbilical cord blood (UCB) and peripheral blood (PB) harvested from patients after mobilization with chemotherapy and/or cytokines. Both contain a large number of lineage restricted and pluripotent progenitor cells and can be expanded extensively ex vivo. Potential clinical applications of gene therapy in such cell populations include correction of inborn enzymatic diseases and support of high-dose chemotherapy by transplanting ex vivo transduced progenitor cells rendered more resistant to cytotoxic drugs. The feasibility and efficiency of retroviral transduction into UCB and PB has been reported recently. PMID- 7475311 TI - Long-term expression of the glucocerebrosidase gene in mouse and human hematopoietic progenitors. AB - Gaucher disease (GD), one of the most common inherited metabolic disorders, is an excellent candidate for gene therapy using hematopoietic stem cells as targets. Animal models have demonstrated the feasibility of introducing the human glucocerebrosidase (GC) gene into hematopoietic progenitors with long term expression using a variety of retroviral vectors. We have previously demonstrated the expression and integration of the human GC gene in mouse hematopoietic progenitors and their progeny 4-8 months post transplant in primary recipients using the retroviral vector MFG-GC. We now demonstrate enzyme expression in peripheral blood lymphocytes of secondary recipients more than 12 months post transplantation. We also show a transduction efficiency of up to 95% in colony forming unit-granulocyte macrophage (CFU-GM) colonies generated from transduced CD34+ cells from a variety of sources, using a centrifugation promoted infection protocol. Transduction has also been documented in long term culture initiating cells (LTCIC) from the same transduced CD34+ cells. These data indicate efficient transduction of mouse hematopoietic progenitors as well as human CD34+ cells using the retroviral vector MFG-GC. PMID- 7475312 TI - Engraftment of SCID mice by human bone marrow hematopoietic cells cultured in vitro: an in vivo model for human gene transfer. AB - Demonstration of the ability of fresh human hematopoietic cells to engraft severe combined immuno-deficient (scid) mice has provided an in vivo assay for expansion and maturation of early human progenitor cells. However, engraftment of cultured hematopoietic cells has been difficult to achieve. We wished to further develop this model as an in vivo assay for efficiency of retroviral gene transfer and expression in the differentiated progeny of adult human bone marrow progenitor cells. Human bone marrow cells were cultured in vitro for six days under conditions suitable for infection by retroviral vectors prior to transfer to irradiated scid mice. Cultured human bone marrow cells introduced by both intravenous (i.v.) and intraperitoneal (i.p.) injection persisted in the bone marrow, spleen and peritoneum of recipient animals up to four weeks after transfer. Following irradiation scid mice receiving cultured human bone marrow cells by either i.v. or i.p. routes demonstrated engraftment of the bone marrow and spleen as determined by the growth of human hematopoietic progenitors in soft agar. By flow cytometric analysis human cells were also detected in the peritoneum of mice receiving cultured human bone marrow cells i.p. These results suggest that the transfer of cultured human bone marrow cells to scid mice with the subsequent engraftment of these cells in the bone marrow, spleen and peritoneum of recipients can routinely occur. This provides an in vivo model for retroviral gene transfer to human cells. PMID- 7475313 TI - Stem cell stimulation in vitro by the dekapeptide (pEEDCK)2: a single-factor alternative for multifactor cocktails. AB - Insertion of foreign genes into cellular DNA requires (at least one round of) DNA replication. Since hemopoietic stem cells do not divide rapidly, numerous semi empirically designed multifactor cocktails have been used to stimulate them. In an attempt to find an alternative to this approach we have investigated the effects of the stem cell stimulatory peptide (pGlu-Glu-Asp-Glys-Lys)2, (pEEDCK)2, on progenitor output in murine long-term bone marrow cultures (LTBMC). (pEEDCK)2 may act by inducing growth factor production in stromal cells. Addition of (pEEDCK)2 to LTBMCs resulted in a three-fold increase in CFU-GM production. For showing an effect of (pEEDCK)2 on primitive hemopoietic cells (long-term-culture initiating cells, (LTC-IC)) LTBMCs were depleted of rapidly dividing progenitors by 5-Fluoro-Uracil (5-FU). LTC-IC survive and repopulate the culture with new CFU GM. (pEEDCK)2 greatly enhanced this process (eight-fold in the second week after 5-FU). Enhanced progenitor production was observed for several weeks even after discontinuation of (pEEDCK)2 additions to the cultures (100-fold, five weeks after 5-FU, three weeks after end of peptide additions). This increase in progenitor production resulted in increased numbers of total nucleated cells. Our results suggest that (pEEDCK)2 may be a useful alternative for multifactor cocktails when proliferation of primitive stem-cell-like cells is required, as in gene therapy and transplantation. Our experiments also indicate that the redox equilibrium between (stem cell inhibitory) monomeric pEEDCK and (stem cell stimulatory) dimeric (pEEDCK)2, which are both endogenous constituents of LTBMCs may play a role in physiological stem cell regulation. PMID- 7475314 TI - Construction of retroviral vectors for targeted delivery and expression of therapeutic genes. AB - Current gene therapy protocols take an ex vivo approach in which cells are removed from a patient, genetically modified and then reimplanted. However this kind of approach is both cumbersome and costly, requiring high tech facilities and is limited to cell types that can be easily cultured. The in vivo delivery of genes by retroviral vectors will greatly facilitate gene therapy protocols of the future. However before in vivo gene therapy becomes a reality a number of problems must be overcome. Ideally therapeutic genes should be delivered only to the relevant cell type and/or expressed in this cell type. Strategies are described that (I) limit therapeutic gene delivery, using pseudotyping or vectors based on retroviruses that show a restricted infection spectrum or (II) limit the expression of transferred genes by inclusion of tissue specific promoters or cis acting regulatory elements. The combination of some of these strategies should permit the construction of novel retroviral vectors that provide safe and targeted in vivo gene transfer. PMID- 7475315 TI - Hematological reconstitution and gene therapy: retroviral transfer of the bacterial beta-galactosidase activity into human hematopoietic CD34+ cell populations and into T lymphocytes derived from the peripheral blood. AB - We report the possibility to transfer marker genes coding for beta-galactosidase activity using retroviral vectors into human peripheral blood CD34+ cells, peripheral blood T-lymphocytes and into the growth factor-dependent human hematopoietic cell line TF-1. Using the MFG-nisLacZ and the FLac vector and various packaging cell lines, we demonstrated retroviral transfer and high expression of a bacterial beta-galactosidase activity induced by the nisLacZ gene or the Sh-ble/LacZ gene. Kinetics of expression of the transgenes were analyzed both in primary cells and cell lines. Absence of cytotoxicity related to the expression of the bacterial beta-galactosidase was assessed in both cell types. These results open interesting prospectives for the use of the beta-galactosidase activity to mark and follow the fate of genetically modified cells isolated from patients prior to reimplantation. PMID- 7475316 TI - Transduction of LacZ gene into leukemia cells using viral vectors of retrovirus and adenovirus. AB - Recent developments in gene therapy techniques enable us to introduce new genetic information into hematopoietic cells. Among the various techniques, we focused on two viral vector systems, one using a retrovirus and the other an adenovirus. By using an adenoviral vector we could transduce and highly express bacterial beta galactosidase (LacZ) gene under the control of the CAG (cytomegalovirus enhancer with chicken beta-actin promoter) promoter in various hematopoietic cells, although the expression persisted for only two weeks. The retroviral vector (MFG) could transduce the LacZ gene into hematopoietic cells almost as well as the adenoviral vector using the repetitive infection protocol. The retroviral system could maintain the expression of transduced cells quite longer than the adenoviral system. Differential use of these two vector systems may be helpful for the gene transduction into various kinds of hematopoietic cells (Lin et al., manuscript in preparation). PMID- 7475317 TI - Retroviral-mediated gene transfer of the human erythropoietin gene into a factor dependent cell line, TF-1. AB - The factor-dependent cell line, TF-1, established from a patient with erythroleukaemia, shows characteristics of immature erythroblasts. Addition of granulocyte-macrophage colony stimulating factor (GM-CSF) to the culture medium is required for long-term growth of the cells. Erythropoietin (Epo) can also be used to sustain TF-1 cells but for only limited periods (approximately a week). Low levels of both growth factors can act synergistically to maintain proliferation for a longer period of time than Epo alone. To eliminate the requirement of exogenous Epo for growth, TF-1 cells were co-cultured with a retroviral secreting cell line containing the human erythropoietin (hEpo) gene and a neomycin (neo) selectable marker. TF-1 cells which exhibited neo resistance (indicating infection by the retrovirus) were then grown in low concentrations of GM-CSF without the addition of Epo. Under these conditions growth of normal TF-1 cells was not sustained. The neo-resistant cells survived for more than 14 days indicating synergy between GM-CSF and the Epo synthesised by the co-cultured TF-1 cells. Radioimmunoassays performed on growth media detected concentrations up to 1 mU/ml of Epo, implying that stable integration of the retroviral vector and expression of the hEpo gene have been achieved. PMID- 7475318 TI - Round table discussion 'gene therapy and society'--summary and conclusions. AB - An international group consisting of scientists, physicians, and one lawyer reported on the problems of gene therapy with regard to society in their respective countries and discussed ethical, legal and scientific aspects of gene therapy. Ethical questions and fear in society were addressed by Dr Odenbach, former Secretary General of the German Medical Association, member of the German Parliamentary Enquiry Commission, 'Prospects and Risks of Gene Technology' and present adviser of the Ethical Committee of the World Medical Association, and by PH Hofschneider, Professor of Virology at the Max-Planck-Institut for Biochemistry, Munich, with a long-time strong interest in medical ethics; legal aspects were addressed by H Hausheer, Professor of Civil Law with many years experience in constitutional law and jurisdiction in Switzerland. Medical and scientific aspects were discussed by R Strohman, Emeritus Professor of Molecular Biology, Berkeley, USA, F Takaku, President of the International Medical Center of Japan and Chairman of the Central Evaluation Committee of the Japanese government for gene therapy, Tokyo and J Goldman, Professor of Leukemia Biology at Hammersmith Hospital, London, and known for this pioneering work in bone marrow transplantation. The discussion was moderated by R Hehlmann, Professor of Medicine at the University of Heidelberg and member of the World Committee of the International Association. PMID- 7475319 TI - Retrovirus mediated gene transfer as therapy for adenosine deaminase (ADA) deficiency. AB - ADA deficiency manifests as a severe combined immunodeficiency with profound T lymphocytopenia. Affected individuals have variable defects of both T- and B lymphocyte function and greatly increased morbidity and mortality caused by frequent viral and bacterial infection. In 1990 a clinical protocol for the treatment of this disease using retrovirus mediated transfer of the ADA gene into peripheral lymphocytes was begun and in 1993 an amendment permitting gene transfer to CD34+ stem cells isolated from peripheral blood or from umbilical cord blood was approved. Five patients have been treated on this protocol and have been analyzed for the persistence of cells containing the transferred gene and for immunologic reconstitution. PMID- 7475320 TI - Gene therapy against retroviral diseases. AB - Eventually, gene therapy may be a valid option for chronic viral infections, including retroviral infections. Human retroviral diseases fit two categories: (1) those that result from a monoclonal outgrowth of a human T-cell leukemia virus type I (HTLV-I)-infected cell, as in the case of adult T cell leukemia (ATL); and (2) those that appear to result directly from virus load rather than monoclonal outgrowth--such as tropical spastic paraparesis/HTLV-I associated myelopathy (TSP/HAM) and human immunodeficiency virus (HIV)-associated acquired immune deficiency syndrome (AIDS). For ATL gene therapy, corrective mechanisms directed at regulatory sequences rather than viral sequences may be most important, though perhaps anti-tax therapy would be useful. For TSP/HAM and AIDS, gene therapy directed to control virus replication may be most useful. For anti retroviral therapy, one may use dominant negative mutants and a variety of other approaches that direct toxins or compete out viral regulatory gene signal sequences. For maximum benefit, such therapy should be directed to different essential genes (eg gag, pol, env, tat or rev) involved in the virus replication cycle and utilize different toxic approaches. A major impediment to the use of gene therapy for AIDS is our inability to transfect a significant fraction of target cells in vivo. Except for reconstituted mice, retroviral systems of animals have been under-utilized as models for gene therapy. Naturally occurring retroviral diseases of cats, goats, horses, and other species provide models for future development. PMID- 7475321 TI - Characteristics of a group of nonnucleoside reverse transcriptase inhibitors with structural diversity and potent anti-human immunodeficiency virus activity. AB - Current thrust in controlling the Acquired Immune Deficiency Syndrome (AIDS) focuses on antiviral drug development targeting the infection and replication of the human immunodeficiency virus (HIV), the causative agent of AIDS. To date, treatment of AIDS has relied on nucleoside reverse transcriptase inhibitors such as AZT, ddI, and ddC, which eventually become ineffective upon the emergence of resistant mutants bearing specific nucleotide substitutions. The Anti-AIDS Drug Screening Program of the NCI conducts and coordinates a high-capacity semi robotic in vitro screening of synthetic or natural compounds submitted by academic, research and pharmaceutical institutions world-wide. About 10,000 synthetic compounds are screened annually for anti-HIV activity. Confirmed active agents are subjected to in-depth studies on range and mechanism of action. Emerging from this intense screening activity were a number of potentially promising categories of nonnucleoside reverse transcriptase inhibitors (NNRTI) with structural diversity but strong and reproducible anti-HIV activity. Over 2500 active compounds were evaluated for their inhibitory activity against a panel of both laboratory and clinical virus isolates in the appropriate established cell line or fresh human peripheral blood leukocyte and macrophage preparations. Out of these, 40 agents could be placed structurally in nine categories with an additional 16 unique compounds that share the characteristics of NNRTI. These NNRTIs were shown to inhibit reverse transcriptase enzymatically using homopolymeric or ribosomal RNA as templates. NNRTIs demonstrated similarity in their inhibitory pattern against the HIV-1 laboratory strains IIIB and RF, and an AZT-resistant strain; all were inactive against HIV-2. These compounds were further tested against NNRTI-resistant HIV-1 isolates. NNRTI-resistant HIV-1 isolates were selected and characterized with respect to the change(s) in the viral reverse transcriptase nucleotide sequence. Also, differential cross resistance or sensitivity patterns to NNRTIs were studied in detail among NNRTI resistant mutants. When tested in combination with AZT, all of the NNRTI's uniformly exhibited synergistic inhibition of HIV-1, suggesting that combination antiviral therapy of NNRTIs with AZT may be therapeutically promising for AIDS treatment. PMID- 7475322 TI - Inhibition of human immunodeficiency virus type 1 (HIV) gag gene expression by an antisense oligodeoxynucleotide phosphorothioate. AB - We have employed a cos-like monkey kidney cell line (B4.14) transfected with plasmids pCMVgag-pol-rre-r (containing the HIV gag and pol genes) and pCMVrev (containing the HIV rev gene), as a model to investigate whether antisense constructs could interfere with specific HIV gene and protein expression. We utilized an antisense construct (GP12A) directed against a non-regulatory region of the HIV genome, to transfect cells that expressed the above-mentioned HIV genes. Our results show that GP12A was able to attenuate levels of relevant HIV mRNA and gag proteins in the absence of cytotoxic effects. PMID- 7475323 TI - V-onc mutation associated with host cell growth in retroviral tumors. AB - In sarcomagenesis in rats infected neonatally with feline sarcoma virus (ST FeSV), v-fes product (P85) was previously shown by us to be a predictive and preventive determinant. In order to explore the part played by P85 in tumor suppression, DNA was extracted from precancerous granulomas and from slow or rapid growing sarcomas induced by neonatal injection of the virus. The v-fes signal from extracted DNA was analyzed by PCR-SSCP. The prototype v-fes gene signal was detected in most lesions and found to be generally amplified in rapid growing sarcomas and in some granulomas. Several v-fes homologs showing varying mobilities in gel were seen in most sarcomas and some granulomas with or without the prototype v-fes signal. In slow growing sarcomas and granulomas induced in hosts that were immunized with ST-FeSV induced syngeneic sarcoma and proved to carry IgG antibody to P85, the prototype v-fes gene was found to be down regulated and v-fes homologs were found to be reduced in number or eliminated. These results suggest that the development of v-fes mutations is associated with the growth potential of cells carrying the v-fes gene, and that host immunity to v-onc product influences the development of virogene rearrangements and results in slow and suppressed growth of tumors caused by neonatal infection with retrovirus. PMID- 7475324 TI - The NCI's AIDS antiviral drug screening program. AB - The Developmental Therapeutics Program (DTP) in the National Cancer Institute has an unique mission by serving as a resource for all academic and private research institutions, the National Institutes of Health in the USA, and the global scientific community in the quest for effective therapeutic agents for treating patients with Acquired Immune Deficiency Syndrome (AIDS) as well as for cancer patients. This communication will concentrate on the developmental therapeutic program on anti-AIDS drug discovery. PMID- 7475325 TI - Modification of M1 cells by exogenous introduction of IL6 gene: a model for gene therapy of acute and chronic myeloid leukemia in mice. AB - A number of recent studies has shown that animals immunized with cytokine secreting primary tumors show resistance against an unmodified tumor cell challenge. In the present study we have evaluated the potential role of IL6, a myeloid differentiation inducing factor, in modifying myeloid leukemia cells, a tumor so far not challenged by this approach. M1 cells transduced with N2 based retrovirus carrying the murine IL6 gene exhibit morphological and functional alterations. Genetically modified M1 cells show significant reduction in the growth constant coefficient and in the ability to form hematopoietic colonies. Flow cytometry analysis demonstrate increased expression of CD11b, CD18, F4/80, FcR and MHC class II, suggesting driven differentiation towards commitment. Transduced cells secrete high level of autocrine IL6 and, upon activation with LPS, high levels of TNF further indicating a functional alteration and differentiation. The insertion of IL6 gene coding for signals of cell activation and improved expression of MHC antigens into myeloid leukemia cells may enable more effective tumor recognition in vivo, and boost the local as well as the systemic immune-mediated anti-leukemia response. PMID- 7475326 TI - Characteristics of ovine and caprine lentivirus infections. AB - A majority of ovine lentivirus (OvLv) infections seen on farms develop after long incubation and a slow progression of disease to death but in nature they may also have short latency and cause acute leukoencephalitis and/or acute arthritis and pneumonia in young kids or lambs with exceptionally high mortality. Histopathologically, OvLv diseases may be characterized by lymphoid infiltration, lymphoid hyperplasia with germinal centers and plasmocytosis in the lungs and/or in the CNS, joints and udder. Lymphoid hyperplasia in lymph nodes and spleen, as well as lymphoid infiltration in the kidneys, are almost always seen in advanced cases. In some cases, it shows similarities to lymphoproliferative diseases that are considered malignant. Alveolar epithelial hyperplasia in the lungs is generally also seen, especially in older goats with caprine arthritis encephalitis virus (CAEV), and proliferation of these epithelial cells may form acine and papillary structures and in some cases are histopathologically indistinguishable from tumor nodules seen in sheep pulmonary adenomatosis. Because of complexities in the host-lentovirus interaction, cell-associated transmission and extensive antigenic and genomic variation among infecting isolates, control of infection or prevention of spread are problematic by traditional methods and exploration of alternative control strategies employing selection and expansion of animals genetically resistant to OvLv or transgenic for certain viral genes, merits consideration. Interestingly, the pure Awassi sheep breed are susceptible to infection but do not develop the disease, as do European breeds or cross-breeds in Israel, ie they are infected but not diseased. It seems that the local Bedouin black goat breed is resistant to infection of CAEV under natural conditions.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7475327 TI - Measuring the self-care practice of children and adolescents: instrument development. AB - PURPOSE: To develop an instrument to measure the self-care practice of children and adolescents. SUBJECTS: 471 students, age 9-18, in 4th-12th grade. METHODS: This methodological study involved item development and testing the instrument for reliability, readability, and construct validity. FINDINGS: Study findings for the 35-item Child and Adolescent Self-Care Practice Questionnaire included a coefficient alpha of .83 and moderate correlations with two self-care agency instruments. Factors corresponding to the self-care requisites in Orem's model were identified using a LISREL confirmatory factor analysis approach. CONCLUSIONS & IMPLICATIONS FOR NURSING: The Child and Adolescent Self-Care Practice Questionnaire can be used in further research to determine how children and adolescents perform as self-care agents. Activities are identified and measured, intervention strategies to improve behaviors can be planned. PMID- 7475328 TI - Self-esteem of pregnant substance abusers. AB - PROBLEM: To explore patterns and levels of self-esteem of pregnant substance abusers. METHODS: A descriptive prospective study to describe the self-esteem of pregnant substance abusers. Subjects (N = 31) were abusing and dependent on three or more legal and/or illegal substances. Subjects were asked one open-ended question regarding their self-esteem, then the Rosenberg Self-Esteem Inventory (RSI) was administered. FINDINGS: Subjects gave 46 responses to the open-ended question. Overall, they used a single word to describe self-esteem. The most frequent response on the RSI was "low" for self-esteem, 23 subjects used positive terms, 20 used negative terms, and 3 reported a neutral term. The RSI confirmed the aspects of low self-esteem. CONCLUSIONS & IMPLICATIONS FOR NURSING: Problems with low self-esteem were evident. Intervention strategies need to be developed to increase self-esteem in pregnant substance abusers. PMID- 7475329 TI - Hardiness and stress: a critical review. AB - TOPIC. A critical literature review of the relationship between hardiness and stress. PURPOSE. To examine literature on the definition and concept of hardiness in both individuals and families, issues of measurement, and the relationship between hardiness and stress; and to identify implications for nursing practice and research. SOURCES. Hardiness as a stress resistance resource, dimensions of hardiness, and the importance of hardiness in family stress theory have been proposed through theoretical report. Instruments to measure hardiness have been developed by Kobasa (1979), Pollock (1986), and McCubbin, McCubbin, and Thompson (1991). CONCLUSIONS & IMPLICATIONS FOR NURSING. The precise nature of the relationship between hardiness and stress remains to be determined. The role of family hardiness in reducing stress, especially in families of children with special healthcare needs, merits further investigation. The potential merits of developing hardiness intervention programs have particular meaning for nursing practice and research. PMID- 7475330 TI - Adaptation to pregnancy and motherhood among subfecund and fecund primiparous women. AB - OBJECTIVE: To examine the effects of infertility treatment on women's ability to adapt to pregnancy and motherhood. METHODS: Fecund (n = 261) and subfecund (n = 103) primiparous women receiving obstetrical care in southeastern Michigan participated in this descriptive, correlational, prospective study. The subjects completed Lederman's Pre-Natal Self-Evaluation questionnaire during the third trimester of pregnancy and Lederman's Postpartum Self-Evaluation questionnaire during the first postpartum appointment. FINDINGS: Mean scores showed that the two groups of women were not significantly different with either adaptation to pregnancy or motherhood. CONCLUSIONS & IMPLICATIONS FOR NURSING: Although subfecund women may experience stress in order to achieve a pregnancy, there do not appear to be any latent effects of this stress on their ability to adapt to pregnancy or motherhood. PMID- 7475331 TI - William Worrall Mayo arrives in America. PMID- 7475332 TI - Value of autonomic testing in reflex sympathetic dystrophy. AB - OBJECTIVE: To attempt to characterize reflex sympathetic dystrophy (RSD) and to determine factors that would predict a response to sympathetic block. DESIGN: We undertook a retrospective analysis on 396 patients with chronic limb pain referred for autonomic testing during a 5-year period. MATERIAL AND METHODS: Clinical endpoints were relief of pain after sympathetic block and a composite RSD diagnostic probability score, based on the clinical attributes of allodynia, protopathia, swelling, and vasomotor alterations. We compared the results of three autonomic tests--resting sweat output (RSO), resting skin temperature (RST), and quantitative sudomotor axon reflex test (QSART). RESULTS: Increased RSO predicted the diagnosis of RSD with 94% specificity, and the specificity was 98% when RSO was considered in conjunction with an abnormal QSART result, the best laboratory correlate (P = 0.003) of the clinical diagnosis. Shorter duration of pain correlated with a warmer limb (P < 0.001), even in the absence of RSD. Response to a single sympathetic block did correlate with the diagnosis (P = 0.031) but correlated most significantly with short duration of pain in the arm (P = 0.001) and laboratory findings in the leg, where increased RST (P < 0.001) and QSART (P < 0.001) were near-perfect predictors of response. CONCLUSION: Sweating abnormalities correlate strongly with the clinical syndrome of RSD, and alterations in RST may be superior to clinical findings in predicting the response to sympathetic block. The findings provide physiologic support for the unproven view of a natural disease progression ("stages"), with better treatment response and a warmer extremity initially. Because certain physiologic trends occur in all patients, general alterations of autonomic function with pain are suggested. PMID- 7475333 TI - Renal artery angioplasty: technical results and clinical outcome in 320 patients. AB - OBJECTIVE: To review the results of percutaneous transluminal renal artery angioplasty (PTRA), including technical success and clinical outcome, at Mayo Clinic Rochester. DESIGN: We retrospectively reviewed our experience with 320 patients who underwent PTRA for stenosis of 396 arteries during a 14-year period. MATERIAL AND METHODS: We reviewed medical records and angiograms of patients who underwent PTRA at Mayo Clinic Rochester between January 1980 and December 1993. The patients were divided into four groups, based on clinical history and angiographic appearance of the stenosing lesion: renal artery atherosclerosis (ASO group), fibromuscular dysplasia (FMD group), previous renal artery bypass or endarterectomy, and renal artery stenosis in a solitary kidney. Technical results of the PTRA were determined by evaluation of angiograms obtained before and after the procedure. Data on patient demographics, blood pressure, antihypertensive medications, and serum creatinine were recorded for the period preceding PTRA, after the procedure, and at last follow-up. RESULTS: All groups had statistically significant reductions in mean arterial pressure and antihypertensive medications after PTRA. The percentage of patients who benefited after renal artery angioplasty was 70% for patients with ASO (8.4% cured), 63% for patients with FMD (22% cured), 53.8% for patients with prior surgical revascularization (23.1% cured), and 91.7% for patients with a solitary kidney (0% cured). No significant overall change in serum creatinine level was noted after the procedure in any group. Complications were comparable to those reported in other studies. The 30 day all-cause mortality rate was 2.2% for the current study, all deaths occurring in the ASO group. CONCLUSION: PTRA rarely leads to a "cure" of renovascular hypertension but provides effective control of blood pressure and decreases the medication requirements in selected patients. PMID- 7475334 TI - Assessing the interaction of irradiation with etoposide or idarubicin. AB - OBJECTIVE: To compare the interactions of two topoisomerase II inhibitors, etoposide and idarubicin, with irradiation. DESIGN: Two mathematical modeling systems were used to assess the interactions. METHODS AND RESULTS: Hamster lung fibroblast cells (V79) were exposed to etoposide or idarubicin for 24 hours before or immediately after irradiation. Post radiation treatment with etoposide or idarubicin resulted in radiosensitization, as demonstrated by a decrease in the mean inactivation dose. Exposure to either drug before irradiation resulted in no radiosensitization. The first mathematical modeling system used was isobologram analysis. This analysis revealed a synergistic interaction if etoposide exposure followed irradiation. The interaction from the combination of irradiation and preradiation etoposide was within the envelope of addivity. Irradiation and postradiation idarubicin exposure also resulted in an interaction within the envelope of addivity, whereas preradiation idarubicin exposure resulted in a slightly less than additive interaction. Next, analyses were performed by the median effect principle. Synergistic interactions were demonstrated for combinations of etoposide and irradiation as well as idarubicin and irradiation. Synergistic interactions were more likely when drug exposure (either idarubicin or etoposide) followed irradiation. Experiments at various ratios of radiation dose to drug concentration showed that the likelihood of a synergistic interaction increased as the drug concentration increased relative to the radiation dose. CONCLUSION: The interaction of irradiation with topoisomerase II-reactive agents should be further explored in human tumor cell lines. PMID- 7475335 TI - Treatment of diffuse coronary artery and vein graft disease with a 60-mm-long balloon: early clinical experience. AB - OBJECTIVE: To report our preliminary clinical experience with a new 60-mm-long angioplasty balloon. DESIGN: We reviewed the results in patients who underwent this type of angioplasty between May and October 1993 at our institution. MATERIAL AND METHODS: The study group consisted of 14 high-risk patients (57% with rest-related angina) and 19 treated coronary segments--52% in native coronary arteries and 48% in saphenous vein grafts (mean age, 9 years). Often, long balloon angioplasty was used in conjunction with laser or transluminal extraction atherectomy. RESULTS: Angiographic success (40% or more visual reduction in diameter stenosis) was achieved in all patients. Intimal dissection occurred in 4 of the 19 treated segments (21%), but each was less than 50% obstructive. No patient required intracoronary stenting. Clinical success was achieved in 13 patients (93%). The one death that occurred was from vein graft distal embolization. At a mean follow-up of 9 months, three patients had required reinterventional procedures, and one patient had undergone a coronary artery bypass operation. No myocardial infarction or death occurred during this period. CONCLUSION: Preliminary clinical experience with a 60-mm-long angioplasty balloon to treat complicated coronary lesions in high-risk patients suggests that, when used alone or in combination with other devices, this new balloon results in high initial success and low complication rates. A larger clinical experience is necessary for accurate assessment of the role of this new balloon catheter. PMID- 7475336 TI - Classification of morphea (localized scleroderma) AB - OBJECTIVE: To classify and describe morphea (localized scleroderma). DESIGN: A review of morphea and its subtypes is presented. RESULTS: The current classification of morphea is incomplete and confusing. As knowledge of the spectrum of disease continues to evolve, the controversy and confusing nature of its multiple subtypes present a challenge for the physician who encounters a patient with this condition. Thus, we propose that morphea be classified into the following five groups: plaque, generalized, bullous, linear, and deep. This classification, based on clinical morphologic findings, will simplify the diagnostic and therapeutic approach. CONCLUSION: Morphea represents a wide variety of clinical entities that seen to be on the opposite end of the scleroderma spectrum from systemic sclerosis. The cutaneous lesions eventually evolve from a sclerotic stage to a nonindurated stage, and residual hypopigmentation or hyperpigmentation follows. The histologic pattern in patients with morphea is similar to that in patients with progressive systemic sclerosis. Although treatment is nonstandardized, hydroxychloroquine sulfate may be beneficial. PMID- 7475337 TI - Venous obstruction due to a distended urinary bladder. AB - Bladder distention is an infrequently reported cause of venous obstruction that may be confused clinically with deep venous thrombosis or congestive heart failure. Urinary symptoms may be minimal or absent. Herein we describe a 73-year old man with unilateral lower extremity edema caused by a distended urinary bladder. In addition, we review the clinical manifestations of 15 previously reported cases of venous obstruction due to urinary retention. Of the 15 patients, all but 1 had painless bilateral lower extremity edema. In most cases, the cause of bladder distention was benign prostatic enlargement. PMID- 7475338 TI - Diagnosis and outpatient management of congestive heart failure. AB - Congestive heart failure causes substantial patient morbidity and mortality in the United States. Symptoms and physical findings can be helpful in diagnosis but have limited sensitivity and specificity. Objective measurement of ventricular function is essential in virtually all patients in whom a diagnosis of heart failure is suspected. Reversible causes of heart failure must be sought. Outpatient management includes education and counseling, emphasis on and assessment of compliance with diet, and pharmacologic treatment. Angiotensin converting enzyme inhibitors are the mainstay of treatment but are underused, and maximal doses are not given apparently because of concern about side effects. Diuretic therapy should be administered only as needed to manage fluid overload. Calcium channel blockers are relatively contraindicated in patients with impaired ventricular function. Patient follow-up should be guided by the results of the medical history and physical examination. Routine serial testing of ventricular function and exercise performance is discouraged. PMID- 7475339 TI - 64-year-old woman with chest pain and presyncope. PMID- 7475340 TI - Chronic diarrhea: the role of magnesium. PMID- 7475341 TI - Dementia: diagnosis and evaluation. AB - OBJECTIVE: To describe an approach to the diagnosis of dementia based on effective assessment methods. DESIGN: We reviewed the literature and summarized the available diagnostic and prognostic studies of dementia that may be useful to the primary-care physician. RESULTS: Although controversy exists about certain aspects of the diagnostic workup, exclusion of potentially reversible causes of dementia is essential. Laboratory studies (for example, for detection of underlying metabolic abnormalities) and neuroimaging of the brain may be useful. The pattern of onset and the temporal course of the disease may suggest a cause and help direct the investigation. Functional losses can substantially impair the patient's ability to live independently. CONCLUSION: Dementia can be mistakenly considered as part of the normal aging process, and diagnosis necessitates a thorough, although not exhaustive, approach. Early identification of dementing illnesses improves the outcome for reversible disease and may also enhance the management of incurable dementias. PMID- 7475342 TI - Management of dementia-related behavioral disturbances: a nonpharmacologic approach. AB - OBJECTIVE: To describe practical nonpharmacologic approaches to dementia-related behavioral problems for enhancement of the function and care of elderly patients with dementia. DESIGN: We reviewed the pertinent medical literature and summarized strategies and available resources for management of geriatric patients with dementia and behavior problems. RESULTS: For optimal care of older patients with dementia, key concepts of related behavior problems must be understood. Agitation and aggression, resisting help with care, wandering, incontinence, sleep disturbance, and emotional lability can become difficult management issues with such patients. In some patients, these disruptions can lead to overmedication and nursing home placement. Herein, practical nonpharmacologic measures for management of behavior problems that arise among community-dwelling and institutionalized elderly patients with dementia are discussed. Attention is directed to the medical, psychologic, environmental, and social factors that may contribute to unwanted behaviors. CONCLUSION: Nonpharmacologic approaches can help ameliorate behavioral problems and assist in the overall care of elderly patients with dementia. PMID- 7475343 TI - Pharmacologic therapies in dementia. AB - OBJECTIVE: To review past and current pharmacologic therapies for cognitive and behavioral problems in elderly patients with dementia. DESIGN: We surveyed the pertinent medical literature and present detailed findings on results of pharmacotherapeutic interventions for cognitive loss and behavioral disorders. RESULTS: Numerous drugs have been used in an attempt to reverse or forestall the cognitive decline associated with dementia. Currently available agents are limited by either lack of efficacy or occurrence of adverse effects. Medications are also used to control undesirable behaviors associated with dementia. Depending on the individual circumstance, such pharmacologic therapies may be of limited utility, some may be harmful, and others may be effective. The use of psychotropic agents in nursing homes is strictly regulated by federal law. For appropriate care of elderly patients with dementia, knowledge of the available drug therapies and their effectiveness is necessary. CONCLUSION: Drug therapies for dementia are associated with adverse effects in the elderly population. Drugs for management of aggressive behaviors associated with dementia should be used only when nondrug strategies have failed. In each patient with dementia, underlying cognitive and behavioral problems must be thoroughly evaluated for selection of the most appropriate drug. PMID- 7475344 TI - Reflex? Sympathetic? Dystrophy? Triple questioned again. PMID- 7475345 TI - Percutaneous renal revascularization. PMID- 7475346 TI - Lung cancer during pregnancy. PMID- 7475347 TI - Lung cancer during pregnancy. PMID- 7475348 TI - Age-dependent changes in response of rat prostatic tissues to isoproterenol and forskolin: changes with sexual maturation in function of G proteins. AB - Developmental changes in the responses of rat ventral prostate to isoproterenol (IPR) and forskolin (F) were studied in relation to the function of beta adrenoceptor-adenylate cyclase system. The response of adenylate cyclase in the tissues to IPR at 10(-7)M and above was steadily enhanced after birth and reached a maximum at 12 weeks, followed by a decrease with age. In contrast, the response of the enzyme to F at 10(-7)M and above was highest at 2 weeks, but thereafter decreased. The changes in the response of the enzyme to IPR coincided with changes in the beta-adrenoceptor density and the binding ability of GTP binding proteins (G proteins) to GTP. The ADP-ribosylation of inhibitory G proteins (Gi proteins) catalyzed by pertussis toxin (IAP) decreased 70% in the tissues from 4 to 8 weeks, and then maintained this level. On the other hand, the ADP ribosylation of stimulatory G proteins (Gs proteins) catalyzed by cholera toxin (CT) increased only 20% in the tissues from 2 to 4 weeks. Thus, the ratio of ADP ribosylation of Gs to that of Gi significantly increased from 4 weeks, reaching a maximum at 12 weeks, but thereafter decreased gradually with age. These changes paralleled those in the function of G proteins and the response of the enzyme to IPR. It is suggested that the rapid and marked decrease in apparent level of Gi proteins in the rat ventral prostate after 4 weeks may have a key role in controlling the function of the beta-adrenoceptor-adenylate cyclase system in the tissues. PMID- 7475349 TI - Oxidative stress and aging in the Mongolian gerbil (Meriones unguiculatus). AB - The objective of this study was to determine if aging in the gerbil, Meriones unguiculatus, is associated with elevation in the level of oxidative stress. Studies were conducted on the brain, heart, kidney, liver and testis of young (3 6 months), adult (15 months), and old (23-25 months) animals. Oxidative damage to proteins, measured as the concentration of protein carbonyls and loss of activity of glucose-6-phosphate dehydrogenase, and to DNA, measured as the concentration of 8-hydroxydeoxyguanosine, increased with age of the animals. There was no appreciable age-related change in the activity of alkaline proteases, which preferentially degrade oxidized protein. Rates of mitochondrial superoxide anion radical and hydrogen peroxide generation also increased with age, most notably in the heart. Antioxidative defenses, measured as activities of superoxide dismutase, catalase and glutathione peroxidase and concentration of glutathione, did not exhibit a uniform pattern of age-related changes. However, when the antioxidative potential of the tissue homogenates was measured as their susceptibility to undergo protein oxidation, in response to experimentally induced oxidative stress, using X-irradiation, tissues of the old animals were significantly more vulnerable than those of the young animals. Results of this study are interpreted to indicate: (i) that the level of molecular oxidative damage to DNA and proteins increases with age, and (ii) that the increased oxidative damage is due to both an an elevation in the rates of oxidant generation and an increase in the susceptibility of tissues to oxidative damage. PMID- 7475350 TI - Age related decline in cytokine induced nitric oxide synthase activation and apoptosis in cultured endothelial cells: minimal involvement of nitric oxide in the apoptosis. AB - Nitric oxide synthase (NOS) activity was enhanced in human umbilical vein endothelial cells (HUVECs) by the combined stimulation with IFN-gamma plus IL 1beta, TNF-alpha and LPS which was accompanied by cell death. DNA analysis of the NOS induced dead HUVECs showed that internucleosomal DNA fragmentation had occurred, suggesting that apoptosis was taken place. The enhanced NO production seemed to be associated with the death of HUVECs, however, both NG-methyl-L arginine (L-NMMA) and nitro-L-arginine (N-arg), inhibitors of NOS, recovered the death of HUVECs by only 16%, suggesting that NO production was minimally involved in the cytokine induced apoptosis of HUVECs. Additional results demonstrated that both the induction of NOS activity and apoptosis in HUVECs declined with in vitro aging, i.e. declined with increasing PDLs of HUVECs, which may explain the decreased immunity during inflammation in aged people. PMID- 7475351 TI - Test-tube simulated lipofuscinogenesis. Effect of oxidative stress on autophagocytotic degradation. AB - Cysteine-stimulated oxidation of a rat liver lysosomal-mitochondrial fraction (LMF) was studied. The process would simulate oxidative stress-related events during the degradation of autophagocytosed material within secondary lysosomes, which may contribute to the formation of lipofuscin or age pigment. Millimolar concentration of cysteine was needed to stimulate LMF lipid peroxidation, measured as thiobarbituric acid reactive substances (TBARS). The amount of endogenous LMF iron was 545 micrograms/l and was enough to initiate peroxidation, probably through the reduction of ferric to ferrous iron by cysteine with induction of Fenton chemistry. Peroxidation could be completely inhibited by the addition of the iron chelator desferal or the antioxidant BHT. A substantial amount of the formed TBARS was associated with trichloroacetic acid (TCA) precipitable proteins. Elevated protein carbonyls was observed 1-2 h after the increase of TBARS. The tryptophan-tyrosine related protein autofluorescence (280/335 nm) decreased sharply during the first few hours of incubation. In contrast, a lipofuscin-type autofluorescence (345/430 nm) appeared only after a few days, suggesting that the latter fluorophore is not an immediate product of protein oxidation. The sequential formation of TBARS, protein carbonyls and lipofuscin-type autofluorescence as well as their dependence on iron and reducing agent add further support to the concept that lipofuscin forms in secondary lysosomes as a result of iron-catalyzed oxidative reactions involving autophagocytosed materials. PMID- 7475352 TI - Evolution of longevity in mammals. AB - Maximum life span is a characteristic of each species, and a diversity of maximum species life spans is seen in each mammalian order. It is argued here that different clusters of related long-lived species evolved independently. A variety of genes could have been involved such as genes for somatic maintenance and genes for resistance to causes of death, including fatal diseases. There are explanations based on natural selection for the long maximum human life span, half of which is post-reproductive. PMID- 7475353 TI - Chromatin structure and the expression of cardiac genes. AB - Actively transcribed genes are more susceptible to nuclease digestion, an observation suggested to reflect an altered state of chromatin organization. It has been hypothesized that exposure or sequestration of chromatin domains is a higher order gene regulatory mechanism. In order to test whether tissue lineage is organized by mechanisms at the level or chromatin structure, three cardiac phenotype-conferring genes (atrial natriuretic factor, myosin light chain-1 ventricular and alpha-tropomyosin) have been assessed for DNase 1 sensitivity in nuclei prepared from tissues of the developing guinea pig. These data have been related to the level of tissue mRNA expression of these genes to ascertain whether the exposed state of genes can occur when transcription is low or undetectable. Although this phenomenon was evident in some cases, the data were not consistent with mechanisms at the level of chromatin structure directing tissue type. PMID- 7475355 TI - Induction and repair of benzo[a]pyrene-DNA adducts in C57BL/6 and BALB/c mice: association with aging and longevity. AB - In this study, we employed the sensitive 32P-postlabeling assay to assess the influence of age on the formation and disappearance of benzo[a]pyrene (B[a]P) DNA adducts in six organs of two different mouse strains with different life spans, C57BL/6ByJ (C57BL/6) and BALB/cByJ (BALB/c). Following a single, intraperitoneal treatment with 50 mg B[a]P per kg of bodyweight, maximum formation of the major B[a]P-derived adduct, trans-(7R)-N2-[10-(7 beta,8 alpha, 9 alpha-trihydroxy- 7,8,9,10)-tetrahydrobenzo[a]pyrene]-yl-deoxyguanosine (BPDE-N2-dG), appeared to be age- and organ-dependent; minor differences were observed for the same organs between the two mouse strains. The maximum formation of BPDE-N2-dG in the various organs from young and old mice differed by a factor of 2-4 and was two- to eightfold lower in organs from old mice as compared to young mice. The removal of BPDE-N2-dG, up to 7 days after the treatment, was apparently age- and strain dependent; non-significant differences were observed for organs within strains at each age studied. In young C57BL/6 mice, which have a greater life expectancy than BALB/c, the rate of disappearance of BPDE-N2-dG was significantly higher in liver and heart as compared to young BALB/c. At the older age a decrease in the rate of BPDE-N2-dG disappearance was observed more frequently, and to a relatively greater extent, in organs from C57BL/6 mice as compared to BALB/c mice. These results are discussed in relation to the differences in life spans and the incidence of pathological lesions between the two strains of mice. PMID- 7475354 TI - Ageing and prostate: age-related changes in androgen receptors of epithelial cells from benign hypertrophic glands compared with cancer. AB - Total and nuclear androgen receptors (AR) were studied from epithelial cells in internal and external prostatic zones in 51- to 86-year-old individuals with benign prostatic hyperplasia (BPH) (n = 68) and prostatic cancer (n = 9). We focussed on the role played by androgens on those processes, despite the fact that at these ages, its secretion has normally decreased. In BPH, the nuclear AR do not change, but total measured androgen receptors rise with age (r = 0.5, P < 0.01). Total or nuclear AR do not correlate with gland volume, despite its increase with age (r = 0.8, P < 0.05). In prostates less than 180 cc in volume, there is a significant correlation between size, serum total testosterone level (r = 0.53, P < 0.05) and prostatic specific antigen (PSA) (r = 0.63, P < 0.05). The amount of nuclear AR in cells from the external zone (infiltrated by cancer or healthy) is two times greater than in those from the internal region. Total receptor content of the external zone cells is also high, but the sample is too small to demonstrate an age dependence. The results suggest that ageing is accompanied by an accumulation of non-nuclear AR in the cytosol, that does not play a role in the development of BPH because the amount of nuclear receptors remains unaltered. The enrichment in nuclear receptors of the external zone cells, independently of the presence of cancer, points to a greater androgen dependence in these cells than in cells of the internal region. PMID- 7475356 TI - Age-dependent enzymuria, proteinuria and changes in renal blood flow and glomerular filtration rate in rats. AB - The time course of urinary excretion of two enzymatic indicators of renal damage, N-acetyl-beta-D-glucosaminidase (NAG) and alanine aminopeptidase (AAP) was measured in female Wistar rats at different ages. NAG and AAP are localized at different sites of the nephron and are released into the urine when kidney damage occurs. Total protein flow, urinary volume and creatinine flow were also determined. In a parallel experiment, the effect of aging on renal blood flow (RBF) and glomerular filtration rate (GFR) was examined in young (1.5-month) adult (3-month) and elderly (20-month) female rats. Clearance following a single injection of [131I]o-iodohippurate (hippuran, OIH) was used for the measurement of effective RBF and as an index of tubular cell function. [125I]Iothalamate (IOT) clearance was used to measure GFR. With advancing age, an increase in NAG and AAP urinary flow appeared. The increases in protein excretion were greater than and previous to those of enzyme excretion. It is shown that absolute RBF and GFR (ml/min) in old rats are greater than in young or adult animals. When absolute RBF or GFR was divided by kidney weight (ml/min/g) no clearance changes appeared in any age group studied; only when clearance was expressed in relation to body weight (ml/min/100 g), a decrease in RBF and GFR was evidenced. This indicates that the rate of increase of both RBF and GFR with age is similar to that of kidney weight and lower than that of body weight. The present findings indicate that urinary markers of renal injury increase with age, whereas GFR and RBF only decrease when expressed as clearance related to body weight. PMID- 7475357 TI - Simulation study for the transition from spindles to spike and wave epileptogenesis. AB - A mathematical model is presented, based on existing anatomical and physiological data, which simulates the behaviour of representative types of cortical cells. It is used to test whether a set of synaptic connections of these cells exists, which, paced by the same rhythmical thalamic input, could produce spindles under normal conditions and spike and wave discharges (SW) under conditions of cortical hyperexcitability. This is possible if the interneurons do not provide recurrent excitatory or inhibitory input on themselves, if the thalamic afferents contact the cortical projecting pyramidal cells through local excitatory neurons, and if the inhibitory interneurons receive input only from the pyramidal cells. The results suggest that an increase of all cortical synaptic actions (both excitatory and inhibitory) is sufficient for the transition from spindles to the first stages in the development of SW discharges in the cortex, whereas the thalamus can be driven to the SW characteristic frequency at the immediate next stages. PMID- 7475358 TI - Velocity profile distribution along an arterial vessel: way to improve detection of stenotic sites. AB - The influence of an arterial stenosis on the pattern of blood velocity spatial distribution is investigated. The blood velocity field in a human femoral artery in a stenotic state is computed by means of numerical simulation. Four distributions of velocity profiles along the vessel are shown, corresponding to four different instants of the cardiac cycle. The shape of the spatial pattern of the velocity profile is strongly perturbed by the stenosis: disturbances are so clear that an easy, precise localisation of the stenotic site is always possible, whatever instant of the cardiac cycle is considered. The reported results prove that a three-dimensional view of the velocity profile distribution along the vessel best emphasises the relevant haemodynamic information from a diagnostic point of view. PMID- 7475359 TI - Model of respiratory sensation and wilful control of ventilation. AB - A mathematical model has been developed that includes sensations of breathlessness and a dynamic CO2 respiratory controller. Breathing sensations, which are represented as a discomfort index, are assumed to depend on arterial PCO2 level, automatic and wilful motor commands and mechanoreceptor feedback. Wilful control is assumed to arise from cortical centres of the brain and is independent of the reflex control system. The bulbopontine respiratory controller produces the automatic motor command, which is determined by chemical and mechanical feedback. Simulations demonstrate how the controller output and breathing sensations change when wilful motor commands disturb spontaneous breathing. Simulations include isocapnic hyper- and hypoventilation and deliberate hypoventilation during CO2 rebreathing. Simulations are compared with experimental data from human subjects. Simulations predict that the discomfort index intensifies when ventilation is either voluntarily raised or lowered from the optimal level; and discomfort is greater when ventilation is lowered than when it is raised at a given level of PCO2. The simulated results agree with those obtained experimentally. The simulations suggest that respiratory drive integration may depend not only on the direct effects of chemical and mechanical feedback, but also on the perceptual consequences of these stimuli. PMID- 7475360 TI - Numerical simulation of instantaneous backflow through central clearance of bileaflet mechanical heart valves at closure: shear stress and pressure fields within clearance. AB - Instantaneous backflow through central clearance of bileaflet heart valves at the instant of closure is investigated. An Edwards-Duromedics valve in the mitral position is employed to measure the transient pressures near the entrance and exit region in an in vitro flow chamber. A region surrounding the clearance is modelled, and two-dimensional quasi-steady-state numerical simulations are performed, with the measured transient pressure difference across the clearance as a driving force for the flow. The results show that pressure difference several times larger than the driving pressure used to close the valve is established across the clearance for about 0.5 ms at the moment of closure. The resulting average wall shear stress is an order of magnitude larger than the turbulent Reynolds stresses reported distal to the valve during opening. A local jump in the shear stress distribution and fall in the pressure distribution are observed at the entrance region. Rounding of the corners in the channel entrance attenuates these spikes. The results of the study indicate that backflow through clearance at closure may be one reason for the haemolysis and thrombosis associated with mechanical heart valves, despite the short duration of the flow field. PMID- 7475361 TI - Assessment of factors affecting the detectability of thermal radiations from the neural system of the brain. AB - The paper describes a technique by which it may be possible to locate and analyse neural activity within the brain by a non-invasive and non-contact method with (in principle) an unlimited bandwidth. The method detects the rapid temperature fluctuations which are believed to accompany individual action potentials and other neural phenomena, and which should be detectable as a noise-like fluctuation of thermal infra-red radiation, with components up to several hundred Hz. It differs from conventional surface thermography in that it ignores changes of mean temperature in blood vessels etc. and transfer of heat to the skull surface by conduction or blood flow. Energy is assumed to travel only as infra red radiation directly from the neural material to the detector, and as such the signal is very heavily attenuated by the water content of the skull and other intervening material. Available data suggest that the viability of the method is considerably reduced by this factor. None the less, the scarcity of reliable data on the temperature fluctuations themselves and the immense advantages of the technique, should it be successfully developed, suggest that it warrants detailed theoretical and experimental study. PMID- 7475362 TI - Beam optics design of compact gantry for proton therapy. AB - A new proton therapy facility for the treatment of deep-seated tumours is being assembled. The proton beam will be applied to the patient under computer control, using dynamic scanning of a focused proton pencil beam to produce a complete three-dimensional conformation of the dose to the target volume. The beam will be applied to the supine patient using a compact isocentric gantry for protons. By combining the scanning of the beam with the beam optics and by mounting the patient couch eccentrically on the gantry, the diameter of the rotating structure can be reduced to 4 m, which is the smallest diameter designed so far for a proton gantry. The paper describes the project especially from the point of view of the optics of the beam transport system of the gantry, including the beam line used to inject the beam into the gantry. PMID- 7475363 TI - Assessment of mass properties of human head using various three-dimensional imaging modalities. AB - Better methods are needed to analyse personal protective devices, such as helmets or facemasks, before they are used in practice. Software tools to quantify three dimensional craniofacial mass properties are developed to improve our understanding of craniofacial impact biomechanics, supplement existing knowledge of osseous structure and provide a comprehensive description of human head morphology. The application of state-of-the-art imaging systems, solid modelling and other software tools are studied to determine the associated errors in mass property estimation by spiral computed tomography, three-dimensional magnetic resonance and optical surface scanning using phantoms and cadaver head studies. Volume, centre of gravity and principal moments of inertia are determined from solid mathematical models for each scanning modality. Landmark-based registration is used to register scans of the same object obtained from the three imaging modalities to a common reference co-ordinate system. Physically and analytically determined mass properties are used as the standard for truth. Although this comparative study does not lend itself to statistical analysis owing to the small sample size, results indicate that any of the three imaging modalities can be used to predict mass properties within the uncertainty of existing methods. Applying these techniques in vivo is practical and feasible. PMID- 7475364 TI - Aspects of silicone rubber as encapsulant for neurological prostheses. Part 2: Adhesion to binary oxides. AB - The paper presents some measurements of the hydrothermal stability of experimental adhesive joints in water at 100 degrees C. The joints are between one adhesive silicone rubber and ten metal or metal-oxide adherends, all combinations of interest to the neurological prosthesis maker. The probable adhesion mechanism is then considered, in the search for some parameter by which the experimental results could have been predicted. Evidence is produced that physical adsorption plays little or no part in the adhesion, but that hydrothermal stability seems to be a function of the adherend ionic charge. In pursuit of this idea, the valency, the Slater potential and the iso-electric point for the surface (IEPS) of the adherend are examined as possible prediction parameters. It is concluded that, at least in neutral and acidified water, the IEPS is a promising predictor, with the benefit that it is experimentally determinable for adherends of unknown composition. The most stable joints seem to occur when the charge density on the adherend has an optimum negative value. PMID- 7475365 TI - Parametric phase-delay estimation of sound transmitted through intact human lung. AB - Sonic noise between 300 and 1600 Hz is introduced into the mouths of 11 healthy adult male subjects at resting lung volume and is detected over the anterior extrathoracic trachea and at three sites on the right posterior chest wall. To overcome the difficulties associated with non-parametric phase unwrapping due to thoracic anti-resonances, the phase delay tau(f) of propagation between the trachea and the chest wall is estimated using a linear parametric ARX-type statistical model with the non-parametric magnitude spectra as a guide. The resulting tau(f) estimates are unambiguous and reliable, and show a clear trend of decreasing tau(f) with increasing frequency, indicating that sound at higher frequencies reaches the chest wall faster than that at lower frequencies. This finding indicates that respiratory sound transmission is highly dispersive, most probably owing to frequency-dependent airway and parenchymal wavespeeds. PMID- 7475366 TI - Identification of noise sources in surface recording of spinal somatosensory evoked potentials. AB - The poor signal-to-noise ratio associated with the acquisition of evoked potentials is a well established fact. The problem is compounded if non-invasive techniques, using surface electrodes, are employed. The paper identifies several sources of noise associated with the acquisition of spinal somatosensory evoked potentials using surface electrodes. In addition, the relative contribution of these sources is determined experimentally for six spinal levels ranging from lower lumbar to upper thoracic. These data will prove useful in the design of digital signal processing schemes such as adaptive noise cancellation, where levels of uncorrelated noise severely limit system performance. PMID- 7475367 TI - Use of finite wordlength FIR digital filter structures with improved magnitude and phase characteristics for reduction of muscle noise in EEG signals. AB - One of the main disturbances in EEG signals is EMG artefacts generated by muscle movements. In the paper, the use of a linear phase FIR digital low-pass filter with finite wordlength precision coefficients is proposed, designed using the compensation procedure, to minimise EMG artefacts in contaminated EEG signals. To make the filtering more effective, different structures are used, i.e. cascading, twicing and sharpening (apart from simple low-pass filtering) of the designed FIR filter. Modifications are proposed to twicing and sharpening structures to regain the linear phase characteristics that are lost in conventional twicing and sharpening operations. The efficacy of all these transformed filters in minimising EMG artefacts is studied, using SNR improvements as a performance measure for simulated signals. Time plots of the signals are also compared. Studies show that the modified sharpening structure is superior in performance to all other proposed methods. These algorithms have also been applied to real or recorded EMG-contaminated EEG signal. Comparison of time plots, and also the output SNR, show that the proposed modified sharpened structure works better in minimising EMG artefacts compared with other methods considered. PMID- 7475368 TI - Synergistic effects of ajoene and the microwave power density memories of water on germination inhibition of fungal spores. AB - The synergistic effects of ajoene and the microwave power density memories of water on germination inhibition of some fungal spores are examined. The study reveals power memory varying different synergistic effects of different concentrations of ajoene on the inhibition of spore germination. PMID- 7475369 TI - Modelling the time-keeping function of the central pattern generator for locomotion using artificial sequential neural network. AB - The paper investigates the ability of a sequential neural network to model the time-keeping function (fundamental frequency oscillation) of a central pattern generator for locomotion. The intention is not to strive for biological fidelity, but rather to ensure that the network obeys the organisational and operational principles of central pattern generators developed through empirical research. The timing function serves to produce the underlying locomotor rhythm which can be transformed by nonlinear static shaping functions to construct the necessary locomotor activation patterns. Using two levels of tonic activations in the form of a step increase, a network consisting of nine processing units was successfully trained to output both sine and cosine waveforms, whose frequencies were modified in response to the level of input activation. The network's ability to generalise was demonstrated by appropriately scaling the frequency of oscillation in response to a range of input amplitudes, both within and outside the values on which it was trained. A notable and fortunate result was the model's failure to oscillate in the absence of input activation, which is a necessary property of the CPG model. It was further demonstrated that the oscillation frequency of the output waveforms exhibited both a high temporal stability and a very low sensitivity to input noise. The results indicate that the sequential neural network is a suitable candidate to model the time-keeping functions of the central pattern generator for locomotion. PMID- 7475370 TI - Evaluation of heart rhythm variability by heart rate or heart period: differences, pitfalls and help from logarithms. PMID- 7475371 TI - Comparison between spot and band electrodes and between two equations for calculations of stroke volume by means of impedance cardiography. PMID- 7475372 TI - Voluntary commands for FES-assisted walking in incomplete SCI subjects. PMID- 7475374 TI - Computer techniques for in vivo determination of geometric properties of human femur and tibia. PMID- 7475373 TI - Magnetic field stimulation of multicellular excitable tissue approximated by bidomain. PMID- 7475375 TI - Acidity of single-part alkoxy-cure silicone rubber DC3140. PMID- 7475376 TI - Generation of geometry of closed human head and discretisation for finite element analysis. PMID- 7475377 TI - Analysis and development of AC biosusceptometer for orocaecal transit time measurements. PMID- 7475378 TI - Heart sound simulator. PMID- 7475379 TI - Dynamic Levkov-Christov subtraction of mains interference. PMID- 7475380 TI - Interface mechanics in external prosthetics: review of interface stress measurement techniques. AB - Instrumentation to measure accurately the stresses at the interface of a residual limb and prosthetic socket has a strong potential for use in prosthetic treatment. As a tool in the clinical setting, the device would allow a clinician to identify sites of excessive loading, information which could then be combined with clinical assessment of skin quality to determine regions of potential skin breakdown. Stress distributions for different prosthetic designs could be compared, facilitating a clinician's judgement to determine the optimal design for a patient. The instrumentation would have additional use in research as an evaluation tool for computer-based finite-element (FE) models. Stump-socket FE models predict stress distributions for proposed socket designs, thus offering advantages over interface stress measurement because evaluation can be conducted before a prosthesis is fabricated or put on an amputee patient. However, FE models must first be proven valid against experimental measurements before they can be considered accurate predictors of interface stresses. Current interface stress measurement techniques are described, with a concentration on a physical explanation of the advantages and limitations with each technique. New emerging technologies are discussed which are instruments that have been described but for which no data collected on amputees have been reported in the literature. The important new features of those technologies are also discussed. PMID- 7475381 TI - Evolution of control system models of ocular accommodation, vergence and their interaction. AB - This paper reviews the evolution of inferential models describing ocular accommodation and vergence, which have been developed using the techniques of control engineering. The models are developed by inferring that the observed behaviour of the ocular systems could be produced by particular types of feedback control systems. The models are subsequently tested and improved by modifications resulting from the comparison of model predictions with physiological experimentation. Current models of accommodation and vergence have had considerable success in describing both the steady-state and dynamic behaviours of the individual systems. However, controversy currently exists among researchers, particularly with respect to the composition of models that describe the interaction of the two systems. PMID- 7475383 TI - Silicon neuron simulation with SPICE: tool for neurobiology and neural networks. AB - The paper deals with computer simulations of 'silicon neurons', which are assemblies of CMOS circuits that generate the equivalents of the ionic currents and of the action potentials of real (biological) neurons. The circuit simulation program SPICE is used to simulate the generation of action potentials by a silicon neuron. Moreover, the equivalent circuits of silicon synapses are described and the behaviours of simple two- and three-neuron networks are analysed. Implications for the areas of neurobiology and formal neural networks are briefly considered. PMID- 7475382 TI - Transducers for foot pressure measurement: survey of recent developments. AB - Recent advances in the development of transducers for the measurement of vertical and shear forces acting on the plantar surface of the foot are reviewed. Barefoot and in-shoe discrete and matrix transducers are reviewed in terms of structure, operation, performance and limitations. Examples of capacitive, piezo-electric, optical, conductive and resistive types of transducer are presented. Where available, the current clinical status is specified. PMID- 7475384 TI - Force-velocity relations of nine load-moving skeletal muscles. AB - The relationship between maximal velocity and load was studied in nine muscles of the cat's hind limb using a technique in which the initial and final muscle lengths are determined by equilibrium of a suspended mass and the muscle's passive and active forces elicited by tetanic stimulation. The maximal velocities of shortening during contraction under each of various loads was used to fit a Hill model using the least-squares method. It was shown that different muscles varied significantly in their ability to generate maximal velocity over a range of loads. The tibialis anterior muscle generate the highest velocity (28.4 cms 1), whereas the tibialis posterior generated the lowest maximal velocity (4.2 cms 1). In general, muscles with predominantly fast twitch fibres and with the largest elongation/shortening range displaced the load at the highest velocities, as compared with muscles with predominantly slow twitch and short excursion range which respond with low velocities. The a/P0 ratio of Hill's equation, which defines the curvature of the force velocity, also varied widely, being most monotonic (0.927) for the soleus and the steepest (0.067) for the extensor digitorum longus, further suggesting that fibre composition is also highly influential on the force-velocity relations of the muscle. PMID- 7475386 TI - Transverse path of ultrasound waves in thick-walled cylinders. AB - The path is examined of a beam of ultrasound through the wall and central cavity of a series of thick-walled cylinders made of polymethylmethacrylate (PMMA) and brass used to simulate the cross-section of the cortical shaft of a long long bone. Water is used as a coupling medium for the external ultrasound transducers and to fill the central cavity. Broadband pulse excitation is employed with two focused 2.5 MHz transducers mounted on a digital calliper. The transmission times of flight are measured on a 100 MHz digital cathode ray oscilloscope. Two distinct pulses of ultrasound were consistently observed at the receiver transducer, the first due to a nonlinear path only in the cylinder wall, and the second due to a linear path travelling directly between the transducers through the centre of the cylinder. The nonlinear path can be explained by refraction at the external surface of the cylinder and diffraction around the central cavity. The nonlinear path is influenced by the relative size of the central cavity. The proposed model for the ultrasound propagation in the cylinder wall gives an excellent correlation with the observed speed and attenuation. The technique can be applied to long bones and has considerable potential as a means of measuring bone quality in human and veterinary medicine. PMID- 7475385 TI - Pressure/flow behaviour in collapsible tube subjected to forced downstream pressure fluctuations. AB - An experimental investigation has been made into the pressure/flow behaviour of a collapsible tube subjected to downstream pressure fluctuations. These downstream pressure waves are observed to be transmitted upstream beyond the point of collapse. The mean flow rate is not significantly affected by the amplitude or frequency of pressure fluctuations. However, the oscillatory flow amplitude is reduced at the higher frequency. The mean flow rate also remains independent of the mean driving pressure. PMID- 7475387 TI - Non-invasive method to detect motor unit contractile properties and conduction velocity in human vastus lateralis muscle. AB - The contractile properties and conduction velocity of motor units are estimated by using surface array electrodes during voluntary isometric contractions of the human vastus lateralis muscle. The subjects develop and maintain sufficient force to steadily discharge a given motor unit, assisted by visual feedback from an oscilloscope. The torque curve developed around the knee joint is triggered by an individual motor unit and averaged. 31 motor units in five subjects are studied. The twitch tension detected ranges from 3 to 27 m Nm with a mean of 12.3 m Nm. The threshold force ranges from 1.88 to 10.12 Nm with a mean of 5.48 Nm, which is 3% of the maximal voluntary contraction. The rise time ranges from 56 to 106 ms with a mean of 83 ms. The mean value of conduction velocity is 4.64 m s-1. The twitch tension is positively correlated to the threshold force (r = 0.839, p < 0.01), but has no relation to the other parameters. It is concluded that the use of non-invasive surface array electrodes provides the contractile properties of motor units and muscle fibre conduction velocity during weak contractions. PMID- 7475388 TI - Method to create small photo-bleached volumes to monitor blood plasma flow in capillaries. AB - A method has been developed to examine the movement of plasma in capillaries using intravital microscopy. Spatial transients in fluorescence properties are instantaneously induced by laser photo-bleach pulses after which the convective recovery can be monitored. The plasma is tagged with fluorescent dyes coupled to bovine serum albumin, which is injected well before the measurements and circulates with the blood stream. A laser beam from an argon laser source, set to emit light with a wavelength of 488 nm, is focused on the illumination field diaphragm and creates a spot in the object plane of the microscope. At low laser power, the laser spot is aimed at a blood plasma gap between red blood cells in a capillary segment, using a steerable mirror. Light sensors, coupled to photo multipliers in the secondary image plane of the microscope, record the light intensity of the moving plasma/dye while the preparation is continuously illuminated with a xenon epi-illuminating set-up. The laser photo-bleach spot is then used to bleach the dye complex within a 5.4 microns segment of the capillary for less than 20 ms. The movement of the bleached plasma bolus is tracked by the photo-sensors, placed sequentially along the capillary. Both dye and red blood cell passage can be detected in the photo-multiplier signals, and the relative velocities of the two blood components can be measured. Measurements reveal that the ratio of transit times between blood plasma and red blood cells is 1.23 (SD = 0.22, N = 18), which is in good agreement with measurements by other techniques. PMID- 7475389 TI - Sonic phase delay from trachea to chest wall: spatial and inhaled gas dependency. AB - A parametric phase delay estimation technique is used to determine the spatial and inhaled gas composition dependencies of sound propagation time through an intact human lung at frequencies of 150-1200 Hz. Noise transmission measurements from the mouth to the extrathoracic trachea and six sites on the posterior chest wall are performed in 11 healthy adult subjects at resting lung volume after equilibration with air, an 80% helium-20% oxygen mixture, and an 80% sulfurhexafluoride-20% oxygen mixture. The phase delay, tau(f), exhibits a bilateral asymmetry with relatively decreased delays to the left posterior chest as compared with the right. The phase delay to lower lung sites is greater than to upper sites at frequencies below 300 Hz; yet the opposite is found at higher frequencies, indicating changing propagation pathways with frequency. There is no measurable effect of inhaled gas composition on tau(f) below 300 Hz. At higher frequencies, changes in tau(f) that reflect the relative sound speed of the particular inhaled gas are observed. These findings support and extend previous measurements and hypotheses concerning the strong frequency dependence of the acoustical properties of the intact respiratory system. PMID- 7475390 TI - Displacement of alveolar macrophages in air space of human lung. AB - The role of alveolar macrophages in the process of the human lung clearance is summarised. Three patterns of alveolar macrophage (AM) displacement on the surface of alveolus are distinguished depending on the loading of the surface with insoluble deposits, i.e. directional, directional with small stochastic noise and purely random. The physical analysis is presented of chemotactic movement and hydrodynamical effects on the residence time of AMs in a geometrical model of the human alveolus. The calculation of exit times from the alveolus is also presented. Calculations show that simultaneous passive and active displacement of AMs loaded with particles reduces exit time of the macrophage by 85%, compared to the case of purely directional movement. When active transport is reduced, due to AM overloading, exit time is determined by the passive transport rate. For reduced surfactant activity, the exit time of AM from the alveolus is the function of its chemotactic activity only and is inversely proportional to AM mobility. The exit time of AMs tends towards infinity when both mechanisms of clearance decay. PMID- 7475391 TI - Individually shaped volume conductor models of the head in EEG source localisation. AB - Inverse solution techniques based on electroencephalograph (EEG) measurements have become a powerful means of gaining knowledge about the functioning of the brain. A model of the head and a potential computation method are necessary to describe the EEG problem mathematically. The generation of realistically shaped three-compartment models of the head is discussed. The isolated problem approach for the boundary element method is applied to develop a fast and accurate numerical solution of the EEG forward problem. Accuracy studies with this approach show that dipole positions can be reconstructed within a distance of 3 mm from the original positions. Inverse simulations indicate that the incorporation of the individual head shape may significantly influence the reconstructed dipole position but not its magnitude and orientation, in comparison with the commonly used three-sphere model. However, the presence of noise in the simulated potential data affects the solutions based on realistically shaped models more than those of the simple three-sphere model. The increased sensitivity of the former models to noise in the data remains a serious drawback for their practical application to EEG source localisations. PMID- 7475392 TI - Catheter-manometer system damped blood pressures detected by neural nets. AB - Degraded catheter-manometer systems cause distortion of blood pressure waveforms, often leading to erroneously resonant or damped waveforms, requiring waveforms quality control. We have tried multilayer perceptron back-propagation trained neural nets of varying architecture to detect damping on sets of normal and artificially damped brachial arterial pressure waves. A second-order digital simulation of a catheter-manometer system is used to cause waveform distortion. Each beat in the waveforms is represented by an 11 parameter input vector. From a group of normotensive or (borderline) hypertensive subjects, pressure waves are used to statistically test and train the neural nets. For each patient and category 5-10 waves are available. The best neural nets correctly classify about 75-85% of the individual beats as either adequate or damped. Using a single majority vote classification per subject per damped or adequate situation, the best neural nets correctly classify at least 16 of the 18 situations in nine test subjects (binomial P = 0.001). More importantly, these neural nets can always detect damping before clinically relevant parameters such as systolic pressure and computed stroke volume are reduced by more than 2%. Neural nets seem remarkably well adapted to solving such subtle problems as detecting a slight damping of arterial pressure waves before it affects waveforms to a clinically relevant degree. PMID- 7475393 TI - Comparison of three running spectral analysis methods for electrogastrographic signals. PMID- 7475394 TI - Dynamic cross-spectral analysis of biological signals by means of bivariate ARMA processes with time-dependent coefficients. PMID- 7475395 TI - Two-dimensional cardiogenic ballistic movements of avian eggs. PMID- 7475396 TI - Magnetic restructuring of water. PMID- 7475397 TI - Portable instrument for accurate measurement of plantar force distribution during dynamic activities. PMID- 7475398 TI - Development of a portable tissue oximeter using near infra-red spectroscopy. PMID- 7475399 TI - Customised optrode holder for clinical near-infra-red spectroscopy measurements. PMID- 7475400 TI - The effects of ownership, operating environment, and strategic choices on nursing home efficiency. AB - This article reports on a study of the labor efficiency of 461 nursing homes located in Pennsylvania. Data envelopment analysis was used to estimate efficiency scores. Tobit equations were estimated for the entire sample and for subsamples consisting of for-profit (FP) and not-for-profit (NFP) nursing homes. The authors found that the major factors explaining efficiency were managerial and environmental characteristics such as ownership, occupancy rate, size, payment source, wage rate, and per capita income, rather than quality characteristics of nursing homes. Analysis of the FP and NFP subsamples suggests that many NFP homes may respond to environmental pressures by increasing their efficiency, whereas FP homes tend to operate at a high level of efficiency irrespective of environmental and regulatory pressures. PMID- 7475401 TI - Does publishing the parameters that trigger review of Medicare claims change provider behavior? Results of the parameter release study. AB - In April 1989, the Health Care Financing Administration stopped publishing the parameters that trigger review of Medicare claims because they thought that knowledge of the parameters would permit providers to schedule visits to avoid review. In this article, the authors report the results of an experimental study where previously unrevealed parameters were revealed to some providers of Medicare services and not to others. The authors find little evidence to suggest that providers schedule procedures to avoid review when they know the parameters. There is some evidence, however, that providers may be responding to the release of the parameters in other ways. PMID- 7475402 TI - Analysis of private health insurance premium growth rates: 1985-1992. AB - The rate of increase in health care expenditures has been a central policy concern for well over a decade, yet little empirical research has been conducted to examine expenditure growth rates. This study analyzed health insurance premium growth rates for a selected sample of 95 insured groups over the period 1985 to 1992. During this time, premiums increased by approximately 150% in nominal terms and by 45% in real terms. The observed rate of growth was not constant over time, however. The most rapid growth occurred during the years 1986 to 1989; thereafter, the rate of increase in premiums declined. Multivariate analysis was conducted to assess the effects on premium growth rates of selected variables representing insurance benefit design features, market competitive factors, insurance system factors, and group-specific factors. In addition to the percentage increase in benefit payments, other factors found to affect premium growth rates were health maintenance organization market penetration, deductible level, the coinsurance rate, and state insurance mandates. Further, this analysis suggests that the insurance underwriting cycle may play an important role in influencing insurance premium growth rates. These results support the belief that health maintenance organization induced competition has potential to control the rate of increase in health care costs. PMID- 7475403 TI - Nursing home survey deficiencies for physical restraint use. AB - New federal regulations, implemented in 1990, severely restrict the use of physical restraints in nursing homes nationwide. This study evaluated the impact of these regulations on North Carolina nursing homes during 1991. An average of 32.6% of facility residents were restrained during that year, and 29.2% of facilities were cited for violation of the restraint regulations. Annual health department surveys of a sample of 195 North Carolina nursing homes were analyzed. The facility proportion of restrained residents was modeled using estimated weighted least-squares regression. A probit model was used to predict if the facility received a deficiency for restraint use. Facility-level factors associated with overall restraint use were the ratio of licensed vocational nurse/nursing assistant staff to residents in the facility, and the overall facility disability level. Factors associated with the receipt of a restraint violation were facility size, direct costs per patient day, the proportion of restrained patients, use of bladder training in less than 3% of residents, and the proportion of residents with organic brain syndrome; variables that were of borderline significance included proprietary affiliation, the proportion of intubated residents, the facility disability level, and the proportion of residents on psychotropic medication. These findings indicate that, despite implementation of these regulations, nearly one third of North Carolina nursing home residents remained physically restrained. The characteristics associated with restraint use and with restraints violations can be used to identify facilities most likely to benefit from assistance and education in reducing physical restraints. PMID- 7475404 TI - Validating the Resource-Based Relative Value Scale cross-specialty alignment. A survey of double-boarded physicians. AB - In 1992, Medicare changed significantly how it paid physicians: instead of basing payment rates on charges, the federal government established a fee schedule based on the Resource-Based Relative Value Scale (RBRVS). Central to the development of the RBRVS was the methodology used to align the work of all specialties onto a single, common scale, termed "cross-specialty alignment." This article presents the methods and results of an alternative approach to cross-specialty alignment. We surveyed physicians who were board certified in both pathology and internal medicine (double-boarded) and asked them about the work of services in both specialties. The results provide a natural bridge between the work of the two specialties. The double-boarded survey results agreed closely with the RBRVS findings. Furthermore, the findings were robust after weighting the estimates by how frequently a physician performs a service or after grouping the physicians by the mix of services in their practice. We conclude that the relative work positions of pathology and internal medicine in the RBRVS have validity. Our findings suggest that a survey of physicians with expertise in multiple specialties is a useful approach for developing cross-specialty linkages for an RBRVS or for validating existing linkages. PMID- 7475405 TI - Conservation of energy, uncertainty reduction, and swift utilization of medical care among the elderly: study II. AB - This study examined age differences in the timing of the decision to seek medical care. The data were obtained from a longitudinal study of 366 community dwelling adults aged 45 to 93 years. Subjects were paired for age, gender, and health status, and both members of a pair were interviewed when either one initiated a medical visit for a new problem. This allowed the authors to examine delay in care-seeking for individuals with new symptoms who did not seek care as well as those who did. Survival analysis was used to test hypotheses respecting age differences for total delay (the time from first noticing symptoms until calling for care) and its two constituent phases: appraisal delay (symptom onset until deciding one was ill) and illness delay (decision one was ill until calling for care). Older persons were expected to be more avoidant of uncertainty and conserving of physical and psychic resources and thus quicker to seek care. The results and findings on reasons for delay support the uncertainly avoidance hypothesis but did not replicate signs of higher levels of avoidance behavior by middle-aged subjects than by older subjects. PMID- 7475406 TI - The Processes, Structures, and Outcomes of Care in Cardiac Surgery. Department of Veterans Affairs Cooperative Study in Health Services No. 5. PMID- 7475407 TI - Development of an interactive data base management system for capturing large volumes of data. AB - Accurate collection and successful management of data are problems common to all scientific studies. For studies in which large quantities of data are collected by means of questionnaires and/or forms, data base management becomes quite laborious and time consuming. Data base management comprises data collection, data entry, data editing, and data base maintenance. In this article, the authors describe the development of an interactive data base management (IDM) system for the collection of more than 1,400 variables from a targeted population of 6,000 patients undergoing heart surgery requiring cardiopulmonary bypass. The goals of the IDM system are to increase the accuracy and efficiency with which this large amount of data is collected and processed, to reduce research nurse work load through automation of certain administrative and clerical activities, and to improve the process for implementing a uniform study protocol, standardized forms, and definitions across sites. PMID- 7475408 TI - The Processes, Structures, and Outcomes of Care in Cardiac Surgery study protocol. AB - Recently, a growing interest has arisen in defining and measuring health care outcomes. Although outcome measures may be used as potential quality-of-care screens, outcomes cannot indicate directly how care might be improved. Thus, the Processes, Structures, and Outcomes of Care in Cardiac Surgery (PSOCS) study was designed to investigate the linkages between the processes and structures of care with risk-adjusted outcomes for cardiac surgery care. Data are being collected on a comprehensive array of risk factors, processes, structures, and outcomes of care at 14 Veterans Affairs Medical Centers for this prospective, observational study. Approximately 6,000 cardiac surgery patients will be enrolled in this study over a 4.5-year period. Patient selection is based on a 6 workday rotating sampling frame with an oversampling of emergent patients. During the study, a register of all patients undergoing cardiac surgery at these centers is being maintained to assess the overall context of patient recruitment. The study will continue to enroll patients through December 1996. Major study end points extend beyond traditional measures of 30-day mortality and morbidity to encompass more innovative intermediate outcome measures, including changes in physical functional status and health-related quality of life. PMID- 7475409 TI - Relationships between patient-related risk factors, processes, structures, and outcomes of cardiac surgical care. Conceptual models. AB - The processes, Structures, and Outcomes of Care in Cardiac Surgery (PSOCS) study involves data collection on a comprehensive array of patient risk factors, processes, structures, and short-term outcomes of care at 14 participating Veterans Administration medical centers. This article summarizes the PSOCS conceptual models that serve as the theoretical framework for analyzing the hypothesized risk-process-structure-outcome relationships being investigated. The PSOCS data set includes more than 1,100 variables related to each patient and more than 300 variables related to provider-specific and facility-specific characteristics. This massive data set presents a formidable analytic challenge. The conceptual modeling process involved four-steps: 1) establishing a vision of the general conceptual model defining the overall risk-process-structure-outcome relationships, 2) developing specific hypotheses or subhypotheses, 3) visualizing a hierarchical set of dimensions and subdimensions, and 4) uniquely assigning each variable collected in the study to a dimension or subdimension for purposes of testing the study's primary hypotheses. A multidisciplinary team participated in this modeling process. The goal of the conceptual modeling process is to identify clearly the actions (ie, the changes in either processes or structures that are linked to risk-adjusted patient outcomes) that can be taken by clinicians, management, and policymakers to improve the quality of cardiac surgical care. PMID- 7475410 TI - Statistical methods and strategies for working with large data bases. AB - This article describes the statistical methods and strategies to be used in establishing the linkages between processes and structures of care with risk adjusted outcomes in a large multicenter Veterans Affairs cooperative study in health services of patients undergoing cardiac surgery. The statistical analyses consist of test involving nine specific hypotheses related to the effect of processes and structures of care on risk-adjusted outcomes. From the statistical point of view, the major obstacles of this study are the need for data reduction and imputation of missing data. The former obstacle is addressed through the use of data-reduction techniques, such as principal components and cluster of variables. The latter is addressed through the use of classic and new techniques for imputation of missing data, such as MISSGEN, principal components for qualitative data, and the expectation and maximization algorithm. Data reduction and imputation of missing data are done with clinically derived variable groups called "dimensions" or "subdimensions." The effect of processes and structures of care is assessed by a two-step process. First, outcomes are modeled using only patient risk factors. The selection of risk factors in the modeling process is discussed in detail. Second, these risk-adjusted outcomes are modeled using one of the nine process or structure subhypotheses. The relationship of the processes and structures of care dimensions and/or subdimensions that are linked to risk adjusted outcomes are identified. PMID- 7475411 TI - Variations in processes and structures of cardiac surgery practice. AB - It is hypothesized that variations in the processes and structures of the selection of patients and the conduct of the surgical procedure may influence risk-adjusted outcome in patients undergoing cardiac surgery. For this reason, the results of the pilot phase of this Veterans Affairs cooperative study were reviewed to determine the variation in the operative practices at six pilot institutions. There were large variations in the percentage of elective, urgent, and emergent cases at each institution, ranging from 58% to 96% elective, 3% to 31% urgent, and 1% to 8% emergent. There was also a tenfold increase in the preoperative use of intra-aortic balloon pump for control of unstable angina, varying from 0.8% to 10.6%. Five of the six centers had accredited thoracic surgical residency programs. There was large variation in the preoperative participation of attending surgeons, from 100% participation at three centers to less than 5% in one. The operation was performed by the attending surgeon in 28% of cases, but this varied from 0% to 100%, depending on the hospital. Cold cardioplegia was used almost uniformly (99%) for myocardial protection; the use of retrograde cardioplegia varied from 2% to 89% among hospitals, and the use of blood cardioplegia ranged from 0% to 100%, with an average of 54% of cases. The use of myocardial temperature monitoring varied between hospitals, from 25% to 99%. The use of the cell saving devices to scavenge shed blood varied from 5% to 99%, and the frequency of the use of banked blood varied from 25% to 65%, depending on the institution. The internal mammary artery was used for 67% patients undergoing coronary artery bypass graft, with a variation between hospitals of 39% to 83%. One hospital used a single cross-clamping technique for the performance of proximal anastomoses in 95% of cases, as opposed to all other hospitals, who used this technique in less than 10% of cases. Aortic venting varied from 58% to 98% and left ventricular venting from 1% to 38%. The use of porcine valves varied approximately 15% in three hospitals to 30% to 40% in the other three hospitals. There were tremendous variations in the duration of operative procedure (5.2-7.3 hours), actual operating time (4.0-5.6 hours), total cardiopulmonary bypass duration (102-146 minutes), and ischemic time (50-87 minutes). The use of inotropic support varied from 41% to 91% between hospitals.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 7475412 TI - Why it is important to demonstrate linkages between outcomes of care and processes and structures of care. AB - This first article of the supplement describes the rationale for the Department of Veterans Affairs Cooperative Study, Processes, Structures, and Outcomes in Cardiac Surgery, which was designed to demonstrate statistically and clinically meaningful linkages between processes and structures of care and the outcomes of that care. United States health care is in an era of great enthusiasm for the use of health care outcomes to assess and improve quality of care. An important reason for this enthusiasm is the concern that processes and structures of care, which traditionally have been selected arbitrarily without valid linkages to favorable outcomes, may not result in the desired outcomes of care. Furthermore, health care outcomes are intrinsic to the definition of quality of care and should be relatively free of preconceived biases about how care should be provided. However, the limitations to outcomes-directed quality improvement have been inadequately recognized. These limitations include the following: (1) mortality, the most commonly used outcome, is usually sufficiently rare, resulting in inadequate statistical power; (2) nonfatal outcomes are much more difficult to measure reliably; (3) outcomes may not be measurable for an extended period of time after the care episode, making linkage to quality improvement inefficient; and (4) patients often desire good processes of care as well as favorable outcomes. A review of the literature found relatively few reports linking processes and structures of care to favorable outcomes. Significant relationships between processes of care and outcomes have been reported for several medical conditions (congestive heart failure, acute myocardial infarction, pneumonia, and stroke) when the patient has been considered the unit of analysis. However, there is a paucity of published meaningful process-outcome or structure-outcome linkages for surgical conditions or for any conditions when the hospital has been the focus of analysis. The authors concluded that quality improvement will proceed most efficiently and effectively if all three elements of Donabedian's quality triad (processes, structures, and outcomes) are used and if the processes and structures chosen have been demonstrated to be associated with desired outcomes of care. PMID- 7475413 TI - Variations in the processes and structures of cardiac surgery nursing care. AB - Nurses play an invaluable role as key members of the cardiac surgery patient's medical care team. Over the last century, the nursing profession has become more independent and autonomous. Despite the widespread use of nursing quality indicators, the effect of nursing-specific processes and structures of care on patient outcomes is unknown. Thus, the Processes, Structures, and Outcomes of Care in Cardiac Surgery (PSOCS) study was initiated, in part, to determine the potential effect of nursing processes and structures of care on cardiac surgery patients' risk-adjusted outcomes. In this article, the authors summarize the key components of nursing structures of care incorporated in the PSOCS study. Nursing process variables were not sufficiently designed into the study to address hypotheses relating nursing care processes to patient outcomes. An analysis of the pilot test data from September 1992 to September 1993 demonstrated potentially important variations between the six pilot centers regarding nursing care provider profiles (eg, educational preparation, specialty certification, and experience levels) and nursing staff ratios (eg, within the surgical intensive care unit). When linked to risk-adjusted patient outcomes, these variations in nursing structure of care may offer important insights toward improving the quality of care of cardiac surgery patients. PMID- 7475414 TI - Evaluating anesthesia health care delivery for cardiac surgery. The role of process and structure variables. AB - Anesthesia care is an integral component of cardiac surgery. Emphasis on cost effectiveness and decreased hospital stay has prompted reevaluation of anesthesia practice. However, the role of anesthesia process and structure variables in relation to patient outcomes is largely unknown. Processes, Structures and Outcomes of Care in Cardiac Surgery is the first epidemiologic study to collect data on anesthesia processes, such as the pharmacologic components of anesthesia and types of cardiovascular monitors used. Structures of care, such as resident staffing, supervision, completeness of documentation, and training and experience of care providers, are also being assessed. Pilot data collected from September 1992 to September 1993 demonstrate substantial variation between the six study sites in selected processes and structures. Despite the near-universal use of narcotic anesthesia as the primary anesthetic technique, variation in the type of opioid and adjuvant benzodiazepine used was observed. Regarding invasive hemodynamic monitoring, most centers used only one type of catheter. Intraoperative transesophageal echocardiography was used commonly at several centers for valve surgery, whereas other centers did not use it at all. Its use during coronary artery bypass grafting was less common. Assessment of the preoperative anesthesia note revealed that coronary anatomy and ventricular function were noted in nearly all instances. However, a clear notation that risks and benefits of anesthesia were discussed was less frequent. Structures related to anesthesia attending staffing, board certification, and experience revealed variation. Some sites had smaller and/or more experienced attending staffs, whereas others had larger and/or less experienced staffs. These pilot findings appear to validate the authors' hypotheses that variations in anesthesia practice are present within the Veterans Affairs system. They suggest that the variable set is robust enough to relate processes and structures of anesthesia care to patient outcome. PMID- 7475415 TI - Self-report measures for assessing treatment outcomes in cardiac surgery patients. AB - Patient self-report measures are increasingly valued as outcome variables in health services research studies. In this article, the authors describe the Functional Status, Health Related Quality of Life, Life Satisfaction, and Patient Satisfaction scales included in the Processes, Structures, and Outcomes of Cardiac Surgery (PSOCS) cooperative study underway within the Department of Veterans Affairs health care system. In addition to reporting on the baseline psychometric characteristics of these instruments, the authors compared preoperative Medical Outcomes Study SF-36 data from the study patients with survey data from a probability sample of the US population and with preoperative data on cardiac surgery patients from a high volume private sector surgical practice. Descriptive analyses indicate that the SF-36 profiles for all of the cardiac patients are highly similar. The Veterans Affairs and private sector patients report diminished physical functioning, physical role functioning, and emotional role functioning as well as reduced energy relative to an age-matched comparison sample. At the same time, however, the Veterans Affairs patients evidenced lower levels of capacity on most of the SF-36 dimensions relative to the private sector patients. PMID- 7475416 TI - An analysis of interobserver reliability and representativeness of data from the Veterans Affairs Cooperative Study on Processes, Structures, and Outcomes in Cardiac Surgery. AB - The authors made some preliminary judgments regarding the reliability and representativeness of the data in the early stages of the Veterans Affairs Cooperative Study entitled Processes, Structures, and Outcomes of Care in Cardiac Surgery (PSOCS). Preliminary PSOCS interobserver reliability and potential patient and site selection bias reported were based on comparisons with identical risk, procedure, and outcome data items collected independently in the Continuous Improvement in Cardiac Surgery Study. PSOCS interobserver reliability for this limited set of variables was good to excellent. At the six pilot centers, there were few important differences between patients entered into PSOCS and those not entered. The 14 Veterans Affairs medical centers that will participate in the full-scale PSOCS study and the 29 nonparticipating centers exhibited similar patient populations. will be valuable. PMID- 7475417 TI - Competition, professional synergism, and the geographic distribution of rural physicians. AB - This study provides a theoretical and empirical investigation of competition and synergism among physicians in rural areas. The results show that rural primary care physicians cluster together rather than distribute themselves evenly. This suggests that public policy makers and rural communities must take an active role to ensure provider availability in all rural areas. There is less clustering among subspecialists. The results also reveal a disturbing negative relationship between young children and physician availability in rural areas. Finally, the results provide strong evidence that the relationship between rural physicians and hospitals is synergistic. PMID- 7475418 TI - Treatment of localized prostate cancer in African-American compared with Caucasian men. Less use of aggressive therapy for comparable disease. AB - The objective of this study is to evaluate differences in treatment for localized prostate cancer in African-American compared with Caucasian men. A cohort of patients was identified from the 1988-1989 Surveillance, Epidemiology, and End Results data base. Data were linked by county of residence to the Area Resource File. The main outcome measures were (1) the receipt of aggressive therapy (radical prostatectomy or external beam radiation) and (2) the treatment modality (radical prostatectomy or external beam radiation). Of 5,786 subjects, 15% were treated with a radical prostatectomy, 29% received external beam radiation, and 56% did not receive aggressive therapy. African Americans were less likely (relative risk 0.46; 95% confidence interval 0.39, 0.54) than Caucasians to receive aggressive therapy. Among those receiving aggressive therapy, African Americans were less likely (relative risk 0.64; 95% confidence interval 0.48, 0.86) than Caucasians to have a radical prostatectomy. Adjustment for patient, disease, and community health care availability characteristics did not change these results. We conclude that African-American and Caucasian men receive different treatments for localized prostate cancer. The reasons for the differences, and their ultimate effect on mortality and quality of life require further study. PMID- 7475420 TI - Physicians' response to length-of-stay profiling. AB - One of the techniques adopted recently by certain hospitals to meet the competitive pressure for reducing costs is physician profiling. Profiling produces periodic reports that compare a physician's resource consumption to a benchmark figure. This study analyzes the effectiveness and implications of one hospital's introduction of physician patient length-of-stay profiling. Data for 24,000 patients treated by 400 physicians in 450 diagnosis-related groups over 42 months were analyzed, including both preprofiling and postprofiling periods. Statistical tests examined whether more physicians achieved the length-of-stay benchmark after profiling was introduced, controlling for physician, disease category (diagnosis-related group), and patient severity level. First, the results establish a significant increase in the percentage of physicians who achieve the length-of-stay benchmark after the introduction of profiling. Second, it was found that physicians who had initially failed to meet the benchmark reduced their patients' average length of stay much more than those physicians who initially achieved the benchmark. Further, reductions occurred primarily at intermediate severity levels, and in diagnosis-related groups with a large economic impact for the hospital. Although the profiling program did achieve the objective of reducing patient length of stay, further improvement may be possible. First, providing different benchmarks or targets for different physicians may extend the improvement to a greater percentage of all physicians involved. Second, an analysis of monthly data on total weighted procedures reveals that the reduction in length of stay resulted in an increase in the number of procedures performed per patient day. This finding suggests that to achieve a reduction in hospital costs and charges, profiling programs should be combined with process improvement initiatives. PMID- 7475419 TI - Predicting hospital costs among older decedents over time. AB - To explain the variation in total real hospital costs among elderly patients who died between 1984 and 1991, a cohort analytic study of the nationally representative sample of elderly subjects included in the Longitudinal Study on Aging (N = 7,527) was carried out. The cohort comprised the subset of 1,778 community-dwelling Americans who were age 70 years and older in 1984, had one or more subsequent hospital episodes, and died by 1991. Hospital charges for 1984 through 1991 were taken from the Medicare Automated Data Retrieval System. Annual hospital charges were adjusted for inflation (restated in 1984 dollars) using the hospital market basket component of the consumer price index. The natural logarithm of aggregated real charges was used in the analysis. Mean total real hospital charges were $24,956 (SD = $27,847). A standard multivariable regression model explained 9.7% of the variance in real total hospital charges. After incorporating additional measures reflecting a respondent's distribution (mean and standard deviation) of comorbidities (as measured by the number of ICD-9-CM codes [truncated at five]) during all hospitalizations in the observation window, the cause of death, and the concentration of charges in the last year of life, the explained variance increased to 29.3%. The most important explanatory factors were the two variables controlling for the distribution of comorbidity, the variable controlling for population density, and the dichotomous variable indicating that the patient's death was related to an acute myocardial infarction. Total real hospital resources consumed by elderly decedents vary substantially. The concentration of resources consumed in the last year of a respondent's life was only marginally significant in predicting total real hospital charges over an 8-year observation window. PMID- 7475421 TI - Variation and quality of self-report health data. Asians and Pacific Islanders compared with other ethnic groups. AB - Variation in the sociodemographic, system, and disease characteristics of Asians and Pacific Islanders compared with other ethnic groups and the quality of standard self-report measures of health and functioning by ethnic groups were examined. Secondary analysis of self-report data from the Medical Outcomes Study, an observational study of adult outpatients who received care in either prepaid or fee-for-service plans, was used to compare 527 Asians and Pacific Islanders to patients in other ethnic groups (16,989 whites, 2,533 African Americans, 1,009 Latinos, and 446 others). These patients all completed a screening questionnaire after visits with a Medical Outcomes Study clinician. Asians and Pacific Islanders were similar to African Americans and Latinos on most sociodemographic and system characteristics, disease status, and risk factors. Ethnicity was a significant predictor of differences in self-reported health. As a group, Asians and Pacific Islanders had better health or health of equal status compared with whites, but were less satisfied and perceived less sharing in the doctor-patient relationship compared with other ethnic groups. However, results suggest that data from standard functioning measures for Asians should be interpreted with caution and that more work is needed to understand better the social and cultural influences on the health of Asians and Pacific Islanders. PMID- 7475422 TI - Homeless veterans' utilization of medical, psychiatric, and substance abuse services. AB - This study focuses on the association between homeless veterans' prior utilization of medical, psychiatric, and substance abuse services and biopsychosocial characteristics reported at admission into a domiciliary care program. Given the large number of veterans in the US homeless population and their health care needs, understanding factors associated with health service use among homeless veterans is significant. Research participants were 429 homeless male veterans who had been admitted to the Domiciliary Care for Homeless Veterans Program site at the West Los Angeles Veterans Affairs Medical Center between February 1988 and July 1992 for treatment of medical, psychiatric, or substance disorders. Results of logistic regression analyses indicated that self-reported need (chronic medical problems, serious psychiatric symptoms, combat stress, alcohol use) and evaluated need for care (evidence of liver dysfunction) were important to veterans' use of health services in the 6 months before program admission. Predisposing social structure factors (education, residential stability, and usual sleeping place) were also significant predictors of service utilization. Overall, need factors were more strongly related to service use. Supplementary logistic regression analyses indicated that comorbidity of need factors deserves attention in understanding homeless veterans' use of services. In conclusion, it is important to attend to predisposing social structure factors as potential barriers to care for homeless veterans. PMID- 7475423 TI - Variation in the use of red blood cell transfusions. A study of four common medical and surgical conditions. AB - This study assessed variation in red cell transfusion practice among adult patients hospitalized with ulcer disease (ULCER), and those undergoing coronary artery bypass grafting (CABG), hip surgery (HIP), or total knee replacement (KNEE). The study design was a retrospective analysis of the 1989 MedisGroups Hospital Comparative Database, and the participants were adult patients presenting for their first admission with ULCER (N = 4,664), CABG (N = 6,812), HIP (N = 4,131) or KNEE (N = 3,042) in the MedisGroups Hospital Comparative Database. Outcome measures were whether a patient was transfused, and the number of units transfused. Logistic regression was used to analyze the decision to transfuse, and linear regression to analyze the number of units transfused. In these analyses, patient characteristics, hospital characteristics, and unique hospital identity were used as independent variables. The percentage of patients transfused was ULCER 50%, CABG 81%, HIP 69%, and KNEE 51%. The range among hospitals in the percentage of patients transfused was ULCER 11% to 76%, CABG 51% to 100%, HIP 36% to 95%, and KNEE 9% to 97%. When only patient characteristics were entered in the linear regression analyses, the R2 values were ULCER 0.33, CABG 0.11, HIP 0.11, and KNEE 0.07. When hospital was added, the R2 increased to ULCER 0.38, CABG 0.29, HIP 0.19, and KNEE 0.20 (P < 0.0001 for the change for all analyses). The results of the logistic regression analyses of the probability of transfusion were similar. There is substantial interhospital variation in the proportion of patients transfused and number of units transfused in the four conditions studied. Patient demographic and clinical characteristics explain a substantial proportion of the variation in transfusion practices for ulcer patients, but little of the variation in the three surgical conditions. PMID- 7475424 TI - Research issues related to optimal oral health outcomes. AB - Patient outcomes form one part of the classic triad used to define quality of care: structure, process, and outcome. Outcomes research in dentistry should focus on global questions such as: What difference does dental care make to a patient? Do patients derive perceptible benefits from their dental care? What dental treatments work the best? Like programs or other courses of action, treatment outcomes are considered on three levels: the policy level, which asks, "Are we doing the right things for our target population, given our resources?" program evaluation, in which specific program objectives are set in terms of achievement or maintenance of certain rates and measures; and the individual patient level, in which specific physiologic, psychological, and sociological measures are used to determine individual treatment outcomes. Each level of evaluation can use treatment outcomes or oral health status measures as evaluation criteria. Clinicians routinely face the challenge of providing "appropriate" care to older veterans--a concept that considers the accuracy of risk assessment and diagnosis as well as treatment outcomes (such as the relative effectiveness and cost-effectiveness of alternative treatment strategies, together with the short- and longer-term physiologic and psychological results). Outcomes research is even more challenging when considering its application in older populations whose physical, social, and psychological health are increasingly interrelated and variable with age. Thus, outcomes research, and particularly research on the appropriateness of care, must consider the values and preferences (utility) of those outcomes for older veterans and their families. Optimal oral health outcomes can be achieved through invigorated oral health services research to better understand patients' preferences related to dental treatment decisions, to define oral health outcomes incorporating these preferences, and to assess the effectiveness of the current dental delivery system and its various components, including the appropriate mix of dental providers and allied health personnel, emerging technologies, and innovative prevention and health promotion strategies. An administrative structure and incentives should be established to support dental health services and outcomes research within the Department of Veterans Affairs (VA). A dental service network with an oversight board could be established to evaluate issues related to treatment outcomes. Alternatively, a Geriatric Research Education and Clinical Center devoted to the improvement of the quality of VA dental care and the oral health care delivery system could be established. PMID- 7475425 TI - Quality and effectiveness issues related to oral health. AB - In recent years, methods have been developed to evaluate effectiveness, cost effectiveness, and quality of oral health services and delivery mechanisms. These evaluation techniques are important to the application of oral health services, because most oral health services have not been adequately evaluated. For many services, evaluation may not be necessary. However, there is wide variation in clinical practices and accumulating evidence that many health practices, although based on reasonable pathophysiologic grounds, are not resulting in their expected health benefits. This has led to the suggestion that all clinical precepts may be questioned. A number of measures are used by researchers to evaluate the quality and effectiveness of oral health services. These include: Effectiveness and appropriateness: Methods available to assess health practices range from clinical observation to strictly controlled randomized clinical trials. Cost effectiveness: Benefit-cost and cost effectiveness analyses. Quality: No tool exists that will single-handedly measure quality. Current methodologies should be based on the following three-dimensional conceptual framework: Structure (evaluation of facilities, equipment, personnel, and organization to deliver care), process (observance of the patient-provider interaction), and outcome (measures of health status and patient outcomes). Little to no information on the quality and effectiveness of oral health delivery systems is available in the United States; the most common system in place is solo, private, fee-for-service practice. Specific questions, conditions, or practices that need evaluation include: Alternative frequencies of oral prophylaxis; Use of fluoride and other remineralization techniques for early decay; Alternative methods to replace missing teeth, including the appropriateness of dental implants; Methods to prevent and treat symptoms of dry mouth; Treatment of the medically compromised patient; Prevention of oral complications of cancer treatment; Appropriateness of new and existing diagnostic technologies; Diagnosis and treatment of oral cancer; and Alternative oral health delivery systems, financing, and expanded responsibilities for allied health professionals. PMID- 7475427 TI - Dental services research in an era of health care reform. Keynote address. PMID- 7475426 TI - Functionally dependent veterans. Issues related to providing and improving their oral health care. AB - This article discusses issues related to the quality and effectiveness of new, emerging, and existing oral health delivery mechanisms and models for the functionally dependent veteran. It describes the particular needs and unique qualities of functionally dependent veterans and enumerates the clinical dental interventions and delivery systems available to this group. Barriers specific to The Department of Veterans Affairs (VA) regarding this population and suggestions for improved access to care are summarized. Suggestions for further research are proposed with the intent that limited resources be utilized so that maximum benefits are provided to the functionally dependent veterans who need care most. What we know: Among the veteran population and the general population, the oldest cohort of elderly (those age 85 and older) are growing faster than any other age group. The rate of functional dependency increases dramatically with age, from about 10% at age 65 to 69 to about 57% after age 85. Functionally dependent elderly are the most likely group to be home-bound or in nursing homes. Compared with independent elderly, the oral status of functionally dependent elderly tends to be poorer. Functionally dependent elderly have higher rates of tooth loss, greater prevalence of denture-related problems, greater prevalence and severity of dental caries, higher prevalence of gingival and periodontal disease, and typically not seen a dentist in at least 5 years. The Department of Veterans Affairs has several advantages over non-VA settings in delivering dental care to the functionally dependent including the presence and availability of dental clinics at VA medical centers and outpatient facilities, the fact that they are free to eligible VA patients, and the greater knowledge base of the dentists. Although VA is effective in delivering oral care to a small percentage of the total population, it is likely that demand will increase and resources exhausted if certain anticipated changes occur (eg, greater demand for dental care and/or changing dental status, such as more elderly with teeth). Questions that need to be answered: What quality of oral care is VA providing to the functionally dependent veteran? What delivery systems are the most effective in providing care to the functionally dependent elderly? Is there a difference in oral health outcomes when dental personnel serve on patient care teams? What barriers exist for delivering quality oral care for the functionally dependent elderly? How can we eliminate or minimize these barriers? PMID- 7475428 TI - Resources needed to enhance and develop health services research to improve the oral health status of the aging veteran in the Department of Veterans Affairs System. AB - Health services research (HSR) in the Department of Veterans Affairs (VA) is one way in which to study the oral health care delivered to aging veterans and to ensure the most effective and efficient methods of meeting their oral health needs. This article details the current resources and infrastructure within VA dentistry that could be used to support a dental HSR agenda, followed by recommendations for improvements. The Department of Veterans Affairs' current situation regarding dental HSR is as follows: Dental health services research within VA needs expansion. Staff dentists have neither the training nor the incentives to conduct dental HSR. Dental clinic administrators are discouraged from being supportive of research due to the high clinical work-load requirements present at most dental services. Dental service affiliations with universities are not structured to support HSR. The HSR personnel outside of the dental services do not have the time nor the inclination to engage in dental HSR. Few recruitment and training initiatives are used to increase the number of VA dental health services researchers. Although funding sources exist that could be used to support dental HSR, the lack of trained and interested research personnel prevents full use. The Department of Veterans Affairs data collection in dentistry are inadequate to support dental HSR. If the required changes occur, VA is uniquely situated to be the largest contributor to dental HSR in the United States and to act as a catalyst for improving dental HSR throughout the nation. Recommendations for improvement of VA dental HSR are as follows: Restructure the Dental Activity System to include detailed diagnostic and treatment data as part of an integrated computerized patient record. Develop and implement a strategic plan with a detailed timeline for increasing dental HSR in VA. Recruit staff dentists with formal research training and provide them protected research time and resources to conduct meaningful research. Develop academic affiliations with centers of excellence in dental HSR. Develop full-time equivalent employee positions dedicated to research at the proposed Veterans Service Areas level. Develop incentives to increase collaboration with non-VA health services researchers in addressing VA issues. Develop dental HSR training programs modeled after the Career Development Program, Dental-Geriatric Fellowship, and Dentist Scientist Programs that will permit VA to become a leader in dental HSR training. Develop mechanisms to ensure adequate and sustained funding for dental HSR projects. Ensure that dissemination techniques are effective and result in improvements in operational effectiveness and quality of care. PMID- 7475429 TI - Overview of research needs and opportunities for improving the oral health of aging veterans. AB - This article summarizes health services research recommendations from and similarities of selected oral health research agendas. Themes of research needs and opportunities emphasize the need for development of more comprehensive measures of oral health and functioning, investigations of the effects of improved access on increased utilization and ultimately on oral health, and studies of the relationship between oral health and general health. In addition, future needs include enhanced technology assessment and studies of science/technology transfer into oral health practice, development of better measures and studies of health outcomes, and investigations of the effects of delivery system variables on costs, productivity, and services. Postconference review and input from The Department of Veterans Affairs providers and patient and policymaker groups is recommended to complete the development of an agenda. Adequate funding, trained research personnel, and a commitment to oral health services research are needed to ensure implementation. PMID- 7475430 TI - Oral health for aging veterans. Making a difference: priorities for quality care. Introduction. PMID- 7475431 TI - The department of Veterans Affairs oral health services and eligibility. AB - The Department of Veterans Affairs (VA) operates one of the largest health care systems in the nation; more than 2.5 million veterans receive care annually. Among the special foci of care within VA is the Dental Service. The Department of Veterans Affairs Dental Service is the largest dental care system in the nation and the largest hospital-based dental care system in the world, receiving more than 1.2 million visits annually. The authors describe the VA dental care system and the larger health care system in which it is embedded. How the system is organized, who is eligible for services, who uses care, and what types of services are used are detailed. Compared with medical care, eligibility for VA dental care is more complex and differs for inpatients and outpatients. Outpatients account for 65% of patient visits and 76% of treatment provided in VA dental clinics. The two largest groups of users are inpatients with compelling medical needs (17%) and outpatients who are totally disabled (24%). A wide variety of services is provided, ranging from diagnostic and preventive care to insertion of crowns, bridges, and removable prostheses. Changes in the nation's health care system mandate introspection by all health agencies. The goal of the VA Dental Service is to become veterans' first choice for dental care. Information needed by VA to best respond to the needs of veterans include the following: (1) reasons for why eligible veterans do not use VA dental care; (2) veterans' oral health needs; (3) definition of optimum care and whether it varies as a person moves from functional independence to dependence; (4) whether VA is providing the most cost-efficient care possible and is best utilizing allied health professions; and (5) whether this care is best provided in a hospital setting. Modifications of data gathering systems are required as a first step to providing the needed information. PMID- 7475432 TI - Research issues related to the oral health status of aging veterans. AB - This article reviews the research on the oral health status of aging veterans and offers recommendations for a research agenda that will improve their oral health and quality of life. Uniform definitions, or a "composite" measure, of overall oral health status would facilitate oral health status measurement. Because such a measure is not available, traditional oral epidemiologic indexes or treatment needs data have been used to identify oral health status in aging veterans. Few studies of national scope have been conducted on veterans. A cost-effective alternative may be cooperative studies within the Department of Veterans Affairs (VA) and other organizations. The Department of Veterans Affairs Dental Longitudinal Study (DLS), begun in 1968, provides the richest data on an aging veteran population. More frequently, VA investigators have examined local or regional veteran populations. These studies have examined oral health status and risk factors associated with oral diseases. Unlike the DLS, most studies have been cross-sectional, and a few studies have examined the effect of systemic disease on oral health and vice versa. Key research agenda items and recommendations include: Development and validation of functional measures of oral health status; Implementation of multicenter, cooperative, descriptive, and analytic epidemiologic studies; Implementation of focused cross-sectional and longitudinal studies on oral health status, treatment needs, and patient outcomes; Enumeration of data sets in the VA system that can be shared with non VA investigators; Implementation of targeted studies of aging populations who have specific comorbidities prevalent in VA; and Use of VA continuum of care to study the relation between oral and systemic diseases. PMID- 7475434 TI - Oral health for aging veterans. Making a difference: priorities for quality care research agenda. AB - One of the major objectives of the conference "Oral Health for Aging Veterans- Making a Difference: Priorities for Quality Care" was to create a research agenda that identifies critical questions and inspires needed research so that dentistry can participate more fully in health care reform. By synthesizing the recommendations made by the eight work groups, an agenda was prepared that suggests that oral health services research should be performed to do the following: 1. Develop a definition of oral health status; 2. Identify the factors that affect access and utilization of VA oral health services; 3. Assess the effects of the use of VA dental services on oral health status; 4. Develop a definition of oral health-related quality of life; 5. Determine the relationships among oral health status, oral health-related quality of life, health-related quality of life, and general quality of life; 6. Examine the outcomes of alternative treatments; and 7. Develop practice guidelines. To implement the proposed research agenda, VA will need to provide the necessary personnel and financial resources. It will need to communicate throughout VA the importance of dental health services research. Most importantly, it must enhance the infrastructure that supports such research, including implementation of new data systems and data collection mechanisms. PMID- 7475433 TI - Oral health, health, and health-related quality of life. AB - Health-related quality of life is a multidimensional concept with five broad domains: opportunity/resilience, health perception, functional states, impairments/diseases, and duration of life. It addresses the tradeoff between how long and how well people live. The health-related quality of life approach has provided greater opportunity for investigation of the interrelations among oral health, health, and related outcomes. The inclusion of patient-driven measures, such as perceptions and functional status, is critical. Oral health-related quality of life measures are being developed and used in research on aging populations. Clinical measures of oral health, perceptions of general and oral well-being, and reported physical, social, and psychological functioning are independent, but correlated, components of overall oral health-related quality of life. An oral health-related quality of life approach benefits 1) clinical practitioners in selecting treatments and monitoring patient outcomes; 2) researchers in identifying determinants of health, tracking levels of health risk factors, and determining use of services in populations; and 3) policy-makers establishing program and institutional priorities, policies, and funding decisions. This overview indicates substantial value in pursuing several recommendations. A theoretical framework from which concepts, measures, and models can be derived must be developed to address oral health, oral health related quality of life, health, and health-related quality of life. Oral health outcomes or states must be identified and classified along some continuum of impairment, function, disability, and opportunity. Indicators of appropriate concepts and domains must be adapted or established. Extended analyses on the relations among oral health, oral health-related quality of life, health, and health-related quality of life should be conducted with use of the Boston VA Normative Aging Study and other appropriate data sets. PMID- 7475435 TI - Access to and patterns of use of oral health care among elderly veterans. AB - Utilization of The Department of Veterans Affairs (VA) dental care system is best understood in the context of the health care system at large. Veterans' use of dental care is apparently influenced by a wide variety of factors that include: barriers to access to non-VA systems (such as a lack of health insurance and the inability to afford non-VA care); ineligibility for VA services (such as absence of a service-connected disability); characteristics of the VA delivery system itself (such as eligibility priorities, geographic distribution of VA facilities, and quality of VA care); and characteristics of the veteran population (such as veterans' illness level, their perceived need for care, and their satisfaction with and perceived quality of VA care). The Department of Veterans Affairs offers a unique opportunity to study the effect of certain delivery system characteristics on utilization behavior and veterans' use of specific dental services. Health services research on the VA oral health care system will 1) provide information useful to improve veterans' oral health and the effectiveness and efficiency of VA dental care delivery and 2) improve our understanding of utilization behavior in large public sector health care systems that are surrounded by private sector health care settings. The following topics should be included in a VA oral care health services research agenda: development of a uniform, system-wide dental care data base; an understanding of the long-term benefits that result from the use of specific dental services and determination of the services that are the most cost-effective; an understanding of the characteristics of delivery systems and of the dental care providers who promote the most appropriate, effective, and efficient care; a better understanding of how VA makes veterans aware of their eligibility; an understanding of the relationship between medical and dental care use in the VA system; and an understanding of the degree to which discontinuity exists in dental care services of veterans. PMID- 7475436 TI - Research issues related to oral health expenditures and financing oral health care for the aging veteran. AB - This article was intended to stimulate research on the financing and cost of oral health care provided to older veterans. Such research is needed as increasing costs and gaps in health care coverage have awakened federal interests in developing national health insurance plans. Although the status of oral health care benefits under such plans remains unclear, plans approaching universal coverage will likely lead The Department of Veterans Affairs (VA) to compete with private providers for patients. Oral health care may be financed as a benefit under a VA-sponsored health plan or financed directly as an appropriation to VA medical centers. This article suggests that research is needed 1) to examine different methods of payment, or rate schedules, that may be used to reimburse providers for the cost of oral health care, 2) to calculate reimbursement rates, and 3) to determine profitability by comparing reimbursement rates with estimates of production costs. The need for such research is underscored by wide variations in operating costs of dental services at different VA medical centers. The cost of oral health care services may include not only the cost of providing dental services but also possible medical care cost "offsets," because patients with better oral health may also experience better overall health status requiring fewer medical services. Finally, the literature suggests that time costs play an important role in rationing care among patients with dental insurance. Studies should be conducted in VA to determine how waiting time, cost of travel, and appointment scheduling affect the use of outpatient dental care services. PMID- 7475437 TI - [Left ventricular hypertrophy in a general hypertensive population in Barcelona]. AB - BACKGROUND: Left ventricular hypertrophy (LVH) is a well known cardiac risk factor. There are no data available as to the epidemiology of this disease in the general hypertensive population in Spain. METHODS: A randomized sample (n = 267) of a general hypertensive population under the age of 80 years was followed in a Basic Health Care Area located in the center of Barcelona, Spain. A Doppler echocardiographic study of the patients was carried out. The patients were considered to have LVH when they demonstrated an index > 134 g/m2 of left ventricular mass in males and > 110 g/m2 in females. The odds ratio (OR) was estimated on presentation of LVH associated with exposure to different factors. RESULTS: A prevalence of LVH diagnosed by echocardiogram was observed in 64% (confidence interval 95% from 58.3 to 69.8%). The independent risk factors associated with the presence of LVH were female sex, age and systolic blood pressure. CONCLUSIONS: Left ventricular hypertrophy is a frequent cardiovascular risk factor in the general hypertensive population in Spain. A systematic search by echocardiography cannot be recommended in primary health care until cost effectiveness studies have been performed. PMID- 7475438 TI - [Level of knowledge on neurology among primary care general physicians]. AB - BACKGROUND: The care of the neurologic patients should be approached from the first level of health care. The aim of this study was to evaluate the level of knowledge concerning neurology that have the general public health care physicians in Catalonia, Spain, and to determine whether the training in family and community medicine (FCM) is valid for the adequate care of neurologic patients. METHODS: A sample of 84 general physicians (GP) from the public health care (PHC), representative of the professionals attending the medical consultations of the Costa de Ponent Catalonian Public Health Care Region (one million inhabitants), was evaluated by a multiple choice test of 36 items which has been previously elaborated and shown to have sufficient psychometric guarantees (objectivity, reliability and validity). RESULTS: The GP from the PHC who had not undergone the Resident Training in FCM demonstrated, as a whole, to have an insufficient level of preparation in neurology (mean = 4.08, DT = 2.5, 69% obtained a score under 5 in a scale from 0 to 10). The physician that have received resident training in FCM showed to be a very relevant factor in the knowledge of clinical neurology (mean = 7.84, DT = 1.1, all scored above 5). CONCLUSIONS: The level of the knowledge of neurology in general public health physicians should be improved, particularly in those who have not received the resident training in family and community medicine. Such a training was found to be useful for acquiring correct preparation in neurology. PMID- 7475439 TI - [Hepatocarcinoma and killer cell activity]. AB - BACKGROUND: Alterations in natural killer (NK) cell activity have been described in patients with different neoplasms including hepatocellular carcinoma (HCC). Nonetheless, the relationship between this activity and the clinical situation of the patients with HCC is not well established. METHODS: A group of 33 patients with hepatic cirrhosis and non treated HCC, a group of 22 patients with only hepatic cirrhosis, and a control group of 31 healthy blood donors were studied. In all the subjects the NK cell activity was determined against the K562 cell line marked with 51Cr by the short duration cytotoxicity test, and the number of NK cells/microliters in peripheral blood was determined by flow cytometry (FACScan) using the anti-CD3, CD16 and CD56 monoclonal antibodies. RESULTS: No statistically significant differences were observed in the NK cell activity or in the number of NK cells/microliters among the three groups studied. In the patients with HCC the NK cell activity was not related to the degree of hepatic function, however the patients with tumors greater than 5 cm in diameter or multinodular tumors showed a significant decrease in NK cell activity without a decrease in the number of NK cells/microliters. CONCLUSIONS: The natural killer cell activity in patients with hepatocellular cancer is not related to subjacent hepatic cirrhosis but is related to tumor size. PMID- 7475440 TI - [The primary care physician, the specialist, and the internist?]. PMID- 7475441 TI - [Antiretroviral treatment against HIV]. PMID- 7475443 TI - [Drug nomenclature (II). International common names in Spain]. PMID- 7475442 TI - [Prosthesis-related thrombosis cleared with fibrinolysis. Usefulness of transesophageal echocardiography]. AB - Thrombosis is an infrequent complication in mechanical valve prostheses carrying a vital risk for the patient. The performance of transesophageal echocardiography (TEE) is high in differentiating the thrombosis from other causes of flow obstruction and is fundamental for establishing treatment. Fibrinolysis is an effective alternative treatment to surgery when the latter carries a high risk. In the case herein reported TEE provided a rapid and precise diagnosis and allowed to prove the resolution of the prosthetic thrombosis following fibrinolytic treatment. PMID- 7475444 TI - [Oncogenes and suppressor genes: apoptosis mechanisms]. PMID- 7475445 TI - [Trends of prevalence of HIV infection in a cohort of parenteral drug users]. PMID- 7475447 TI - [Why are patients patients admitted at the hospital as emergencies?]. PMID- 7475446 TI - [Evaluation of adequacy of admission and stay in patients admitted at a tertiary hospital through the emergency service]. PMID- 7475448 TI - [Herpetic encephalitis: diagnostic value of the polymerase chain reaction in cerebrospinal fluid]. PMID- 7475449 TI - [Maniac episode associated with stage C3 HIV infection]. PMID- 7475450 TI - [Spanish public opinion concerning organ donation and transplantation]. AB - BACKGROUND: Information, beliefs, attitudes and the motivations of the Spanish population regarding organ donation and transplantation were studied to facilitate possible programs directed at the promotion of organ donation. METHODS: The Psychosocial Aspects of Donation Questionnaire (PADQ) was applied by personal interview to a representative sample of the Spanish population (n = 1,288). RESULTS: Sixty-five percent of the population was found to be in favor of donating their own organs while only 6.1% had an organ donation card. The main reasons inducing those questioned to be donors were solidarity (47%) and reciprocity (29%). The fundamental reasons hindering donation were not knowing how to be a donor (34%) and the fear of the possibility of an only apparent death (24%). Ninety-three percent would donate the organs of a dead relative if they knew that this person was in favor of donating. Only 52% would donate if they did not know the will of the departed relative. The younger subjects with a higher level of education, higher socioeconomic status, and better informed evaluated donation and transplantation favorably and those with direct experience showed a more favorable inclination to the donation of their own organs. CONCLUSIONS: Educational programs directed at the population should provide information on the efficacy and the profitability of transplantation presenting real cases and emphasizing the quality of life obtained by the receptors. Incentives to organ donation should be directed at reasons of solidarity and reciprocity. Family debate of the matter should be promoted as should the establishment of clear and accessible procedures for the obtaining of a donor card and thereafter the communication of the holders as to their decision to their respective relatives. PMID- 7475451 TI - [High-dosage chemotherapy and the autologous transplantation of peripheral hematopoietic progenitor cells in breast cancer: the initial results, analysis of toxicity and the necessary support means]. AB - BACKGROUND: In the last years high dose chemotherapy (HDC) schedules have been developed with autologous bone marrow transplantation (ABMT) which are very effective in breast cancer. Expectation has been raised concerning the cure of a subgroup of patients with metastatic breast cancer and the improvement of prognosis in high risk stages II and III. METHODS: CTCb (cyclophosphamide 6 g/m2, thiotepa 500 mg/m2 and carboplatin 800 mg/m2) was administered with autologous peripheral hematopoietic progenitor cells transplantation (TACPHP) and granulocytic colony stimulating factor (G-CSF) 5 micrograms/kg/day to 27 patients with breast cancer: 9 in stage IV in complete remission, 12 in stage II with > or = 10 affected lymph nodes and 6 in stage III. RESULTS: No toxic deaths were reported. The median time to achieve > or = 0.5 x 10(9) neutrophils/l was 8 days, to > or = 20 x 10(9) platelets/l 9 days and to > or = 50 x 10(9) platelets/l 12 days. Fever was observed in 85% of the patients although its median duration was of only one day. Extrahematologic toxicity was moderate with grade III nausea/vomiting in 48% of patients, grade III mucositis in 22%, grade III hepatitis in 19%, and grade III diarrhea in 4%. No grade IV toxicity was observed. The median follow-up is still short (10 months, range: 2-25). All the patients maintain normal hematologic peripheral blood counts and only 4 (in stage IV) have relapsed. CONCLUSIONS: The slight extrahematologic toxicity observed in the high dose chemotherapy with cyclophosphamide, thiotepa and carboplatin, and the rapid hematologic recovery provided by the TACPHP and G-CSF allow the above schedule to be administered with moderate toxicity and no mortality. This low toxic profile leads to the possibility of future trials with this chemotherapy schedule in other subgroups of patients with breast cancer. PMID- 7475452 TI - [Nontraumatic rhabdomyolysis: its etiology and the predictive factors of acute kidney failure]. AB - BACKGROUND: The aim of this study was to determine the causes of non traumatic or medical rhabdomyolysis (RM) and analyze the clinical and biologic characteristics, as well as the predictive factors of acute renal failure (ARF). METHODS: Fifty-nine patients with RM admitted to the Internal Medicine Department over an 18-month period were prospectively studied. The patients were classified into etiologic groups. Anamnesis, physical exploration and general analytical studies including creatinkinase, myoglobin, aldolase, magnesium and proteinogram were performed. Urine sodium was determined in patients with ARF> RESULTS: Myalgias in 14 patients (23.7%) and a decrease in strength in 11 patients (18.6%) were the main clinical manifestations observed. As a sole cause, convulsion was the most frequent manifestation in 11 patients (18.6%) followed by muscular compression in 10 patients (16.9%). In the plurietiologic group the most frequent manifestation was muscular compression in 5 patients (16.9%) followed by alcoholism in 4 patients (6.7%). No differences were found in either the clinical manifestation or the biology of the RM among the different etiologies. Eight patients presented ARF (13.5%) with one patient requiring dialysis, and good response being observed with conservative treatment in 5 patients. On multivariant analysis the statistically significant ARF predictive factors were: age over 70 years (p = 0.003), nonketotic hyperosmolar diabetic coma (p = 0.01), previous dehydration (p = 0.001) and initial uric acid levels greater than or equal to 6 mg/dl (p = 0.002). CONCLUSIONS: The clinical expression of rhabdomyolysis was small with the most frequent manifestations being convulsions and muscular compression. The predictive factors for acute renal failure were advanced age, nonketotic hyperosmolar diabetic coma, dehydration, and hyperuricemia. PMID- 7475453 TI - [The generosity of Spaniards and organ transplants]. PMID- 7475454 TI - [The "client" paradox in the health services]. PMID- 7475455 TI - [Drug nomenclature (III). A proposal for the spelling standardization of common international names and their adaptation from English to Spanish]. PMID- 7475456 TI - [Fever and pulmonary cavitation in a 22-year-old woman with recurrent pneumothorax]. PMID- 7475457 TI - [Defects in the prediction of results by APACHE II. An analysis of the errors in predicting the mortality of patients in a critical state]. PMID- 7475458 TI - [Specialized education in emergency medicine]. PMID- 7475459 TI - [Acute kidney failure due to Corinarius orellanus]. PMID- 7475460 TI - [Can the "double blind" be recognized before the identifying codes are revealed?]. PMID- 7475461 TI - [The efficacy of sulbactam-ampicillin in a urinary infection due to Acinetobacter calcoaceticus var. anitratus]. PMID- 7475462 TI - [Transjugular intrahepatic portosystemic shunt and intravascular hemolysis]. PMID- 7475463 TI - [Type 2 human immunodeficiency virus infection (HIV-2) in Spain: analysis of the first 50 cases. Spanish Group for the Study of HIV-2]. AB - BACKGROUND: Infection by the human immunodeficiency virus type 2 (HIV-2) is endemic in West Africa where it is responsible for many AIDS cases. HIV-2 has been described in subjects in other countries, mainly in African immigrants, although it may also be found in natives. The first cases of HIV-2 infection were identified in Spain in 1988. METHODS: The clinical, epidemiologic and virologic characteristics of the subjects with HIV-2 infection identified in Spain up to November 1994 are described. RESULTS: Fifty people have been diagnosed with HIV-2 infection, being found mainly around the large urban centers (20 cases in Madrid and 16 in Barcelona). All the cases were African immigrants except for 10 Spaniards (20%), 6 of whom had acquired the infection in endemic areas while the other 4 (one prostitute from Barcelona and 3 bisexual males from Guipuzcoa) had acquired the infection in Spain. Heterosexual transmission was implicated in all the cases except in 6 subjects (4 bisexual males, one case of probable intravenous transmission and one vertically infected child). Eight (all Spanish) patients have developed AIDS and 5 have died to date. At least 10 other subjects have a CD4+ lymphocyte count lower than 0.2 x 10(9)/l, one being the only pediatric patient (6-year-old male). CONCLUSIONS: HIV-2 infection is present in Spain, although there is no evidence for a rapid dissemination of the virus outside the collective of African immigrants. The Spaniards with HIV-2 infection represent 20% of the cases having contracted the virus through sexual relations with natives from endemic areas. PMID- 7475464 TI - [HTLV-I infection in Spain. Analysis of 24 cases identified until November, 1994. Spanish Group for the Study of HTLV-I/II)]. AB - BACKGROUND: HTLV-I is a human retrovirus which has been implicated in the genesis of tropical spastic paraparesia (TSP), adult T-cell leukemia (ATL) and some patients with uveitis, subacute arthropathies and lymphocytary alveolitis. The virus is endemic in some zones of the Caribbean countries, Japan, subsaharian Africa, Middle East and Melanesia. Given that HTLV-I is transmitted by similar routes as HIV, anti-HTLV-I antibody screening is carried out in blood donors in some countries. METHODS: The clinical, epidemiologic and virologic characteristics of the patients with HTLV-I infection identified in Spain up to November 1994 are described. RESULTS: Twenty-four Spanish residents have been identified with HTLV-I infection including 16 Spaniards and 8 immigrants from endemic areas. Thirteen (53%) are males and 11 (47%) females. Most of the persons born in Spain (12/16; 75%) have lived in endemic areas or have maintained sexual relations with natives of them. Four patients were diagnosed with TSP, three with ATL and another with lymphomatoid granulomatosis and angiocentric T-cell lymphoma. The remaining patients were asymptomatic at the time of diagnosis. Two HTLV-I carriers were identified on blood donation. CONCLUSIONS: HTLV-I infection is present in Spain being found in Spanish natives and, to a lesser degree, in immigrants from endemic areas. It is therefore recommendable to analyze the cost benefit of anti-HTLV-I screening in blood donors. PMID- 7475465 TI - [HTLV-II infection in Spain. Analysis of 113 cases identified until November, 1994. Spanish Group for the Study of HTLV/I/II]. AB - BACKGROUND: HTLV-II is a human retrovirus considered to be responsible for the genesis of some lymphoproliferative and neurologic syndromes. The virus is endemic in some Amerindian and African tribes as well amongst injecting drug addicts (IDA) in North America and Europe. METHODS: The clinical, epidemiologic and virologic characteristics of the patients with HTLV-II infection identified in Spain up to November 1994 are described. RESULTS: One hundred thirteen subjects have been identified with HTLV-II infection in Spain with 4 being African immigrants residing in Madrid and the remaining being IDA from other European countries. Most were males (94/113; 83%). All were IDA except six (5 had acquired the infection by sexual contact and one by blood transfusion). Most of the IDA infected with HTLV-II were coinfected with HIV-1 (93/113; 83%). No patient showed clinical manifestations attributable to HTLV-II infection although one drug addict male coinfected with HIV-1 and HTLV-II developed a non inflammatory proximal myopathy. CONCLUSIONS: Infection by HTLV-II is present in Spain and affects with preference to injecting drug addicts. It has been shown to be of growing incidence with a current global prevalence of 2% in IDA. PMID- 7475466 TI - [Retrovirus, drugs, and sex: a reflection beyond medicine]. PMID- 7475467 TI - [Clinical practice guidelines: is their development worthwhile?]. PMID- 7475469 TI - [Blood markers in the prenatal diagnosis of Down's syndrome: the triple test]. PMID- 7475468 TI - [Selective pulmonary vasodilator effect of inhaled nitric oxide in a patient with pulmonary thromboembolism]. AB - The increase in pulmonary vascular resistences in acute pulmonary thromboembolism (APT) is the consequence of anatomical obstruction and pulmonary artery constriction. The administration of inhaled nitric oxide (NO) may be therapeutically useful in acute pulmonary hypertension by APT given its limited vasodilator effect on pulmonary circulation. A patient with APT in whom this selective vasodilator effect was observed is presented. The authors suggest that the administration of inhaled NO may be a potentially beneficial coadjuvant therapy in acute pulmonary hypertension induced by APT. PMID- 7475470 TI - [Hepatopulmonary syndrome: a new concept. A new treatment?]. PMID- 7475471 TI - [Value of cerebral SPECT with thallium-201 in the diagnosis of primary cerebral lymphoma in AIDS]. PMID- 7475472 TI - [Castleman's disease: histomorphologic association with Hodgkin's disease]. PMID- 7475473 TI - [Ageusia caused by terbinafine]. PMID- 7475474 TI - [Synchronous colonic adenocarcinoma and hypernephroma]. PMID- 7475475 TI - [Maculopapular exanthema caused by ritodrine, accompanied by transaminase increase]. PMID- 7475476 TI - [Usefulness of serial cranial MR in isolated myelitis for the diagnosis of multiple sclerosis]. PMID- 7475477 TI - [Epidemiology of hip fracture in Asturias]. AB - BACKGROUND: Hip fracture is the complication with the greatest medical repercussions in osteoporosis. The aim of this study was to know both the incidence of osteoporotic hip fracture in Asturias, Spain and the immediate evolution of these patients. METHODS: The clinical histories of all the patients admitted for hip fracture in 1992 in two Asturian public health care areas were reviewed. Hip fractures in patients under the age of 45 years or those occurring because of other diseases (metastasis, serious injury) were excluded. RESULTS: In the health care areas studied in 1992 there were 283 osteoporotic hip fractures with an incidence of 219.6 fractures/10(5) inhabitants/year in those over the age of 50 years. The incidence in women over 45 years of age was 3-fold greater than that in men of the same age. The mean age of the patients was 80.2 +/- 8.9 years. The incidence in people over the age of 50 was greater in urban (266/10(5) inhab./year) than in the rural areas (185.7/10(5) inhab./year) (p < 0.001). Ten percent of the patients were residing in old age residences or hospitals at the time of the fracture. Eighty-one percent required surgery. Home was the site of the fracture in 84% of the cases with no seasonal variation being observed. The most frequent intrahospitalary complications observed were infections (15%), cardiac or respiratory disturbances (18%), and confusion (8%). The mean hospital stay was 26.5 days with acute hospitalary mortality of 5.6%. Nineteen point seven percent of the patients were transferred to a center for chronically ill people. The total cost of the hospital care for the hip fractures in this study exceeded 311 million pesetas in 1992. CONCLUSIONS: The epidemiology of osteoporotic hip fracture in Asturias, Spain, follows a similar pattern as that found in other Spanish regions. It was found to be more frequent in the urban than in the rural areas. Hospitalary mortality of these patients is partly determined by age and the number of complications which developed during hospital stay. The impact on the patients with osteoporotic hip fracture and on the economic resources destined to their attention justifies the development of osteoporosis and fracture prevention programs. PMID- 7475478 TI - [Clinical characteristics of the white coat hypertension]. AB - BACKGROUND: White coat hypertension (WCH) is a relatively prevalent clinical situation (around 20% of the hypertense population). The aim of this study was to analyze the clinical characteristics of these patients. METHODS: A descriptive transversal study was carried out in general medicine consultations in a unrandomized sample of hypertensive patients receiving pharmacologic treatment. Twenty-four hour ambulatory blood pressure monitorization (ABPM) (SpaceLabs 90202 90207) was performed following three resting blood pressure (BP) determinations carried out during the visit at 5-minute intervals by mercury sphigmomanometer. WCH was defined as: BP during the consultation > 140 mmHg (systolic) or > 90 mmHg (diastolic) with a mean daily blood pressure by ABPM < 140/< 90 mmHg. RESULTS: One hundred sixty-four patients (53% males) with a mean age of 48 years (SD 15.8) and a mean time of high blood pressure of 36 months (SD 51.4) were studied. Seventy-one patients (43%) fulfilled WCH criteria. WCH was significantly associated with females (p < 0.01), hypercholesterolemia (p < 0.05), lower involvement of target organs (p < 0.05), greater nocturnal tolerance to ABPM (p < 0.05) and lower mean nocturnal blood pressure (p < 0.001). CONCLUSIONS: The prevalence of white coat hypertension is elevated. Most of these patients are females, have fewer target organ lesions that the remaining hypertensive patients although dyslipemia is more frequently observed in these cases. PMID- 7475479 TI - [Ancylostomiasis and strongyloidiasis: clinico-epidemiologic comparative study of travelers coming from endemic areas]. AB - BACKGROUND: The increase in immigration and adventure travelling by tourists generates a population susceptible to have a type of helminthiasis which we are not familiar with. The clinical-epidemiologic characteristics of different groups of patients with this disease are presented. METHODS: A descriptive retrospective longitudinal study was carried out. The clinical-epidemiologic and imaging laboratory data of 56 patients with ankylostomiasis and strongyloidiasis observed in the Department of Tropical Medicine of the Hospital Clinic i Provincial, in Barcelona, Spain, were collected. RESULTS: Ankylostomiasis and strongyloidiasis represent 2% of the global diseases attended. The risk factor the "use of open footwear" was present in almost all the cases. Among the immigrants this parasitosis may become manifest up to 10 years after the last contact. The digestive form and the accidental finding were the most common forms of presentation among the tourists and immigrants, respectively, with diarrhoea being the most frequent manifestation among the tourists (p = 0.008). Forty-five percent of the patients presented polyparasitism, being more frequently observed among the immigrants (p = 0.025). Anemia was found among 44% of the immigrants while this was not observed in any of the tourists (p = 0.012). No clinical relapse was observed with treatment with mebendazol and thiabendazol. CONCLUSIONS: A clinically different behavior was observed between the two groups of travellers. Although infrequent, the clinical importance ot its diagnosis is based on avoiding severe complications in immunosuppressed individuals and preventing anemia in patients with several diseases. PMID- 7475480 TI - [Ardystil syndrome]. PMID- 7475481 TI - [Research ethics committees: a necessary complication]. PMID- 7475482 TI - [Urgent letter from Haiti]. PMID- 7475483 TI - [Sepsis: pending subject for the XXI century]. PMID- 7475485 TI - [Strategies for universal vaccination against hepatitis B]. PMID- 7475484 TI - [Acute renal insufficiency and respiratory failure in a 65-year-old male 3 years after renal transplantation]. PMID- 7475486 TI - [Disseminated cryptococcosis with lymphatic and bone marrow involvement in a patient with HIV infection]. PMID- 7475487 TI - [Rapid cycling following the administration of corticoids to a patient with bipolar disorder]. PMID- 7475488 TI - [Lyme borreliosis in children: double infection or atypical course]. PMID- 7475489 TI - [Panic disorder secondary to anaphylactic reaction]. PMID- 7475490 TI - [The Spitzer's quality of life index: validated?]. PMID- 7475491 TI - Genital tract infections in women. AB - The future holds promise of new, more accurate diagnostic tests; more effective therapies; and development of better barriers and eventual vaccines. GTI is, however, and for the foreseeable future will continue to be a major health care problem for women, requiring careful surveillance, management, and ongoing education for both patient and provider. PMID- 7475492 TI - Psychosexual gynecology. AB - The psychosexual health of the female patient not only is affected by her overall gynecologic well-being but also may be greatly influenced by cultural myths and attitudes, psychological health, and social support. For primary care physicians to incorporate the psychosexual issues of a woman into her overall medical care, there must exist a sensitivity to the patient's needs, an atmosphere for open discussion regarding sexual and psychological concerns, an office staff to provide appropriate medical support, and the availability of experienced specialists in cases in which referral is necessary. The quality of the physician patient interaction as well as the degree of empathy shown to women with gynecologic problems may exacerbate or improve psychosexual outcomes. PMID- 7475493 TI - Diagnosis of early pregnancy and early complications. AB - First-trimester pregnancy disorders are common and range from minor to life threatening conditions. Many of these disorders are encountered by the primary care physician and may be appropriately managed in that setting. Understanding these disorders provides a cost-effective framework for their management. PMID- 7475494 TI - Menopause. AB - The use of naturally occurring sex steroids in humans represents faithful replacement of the steroids lost at ovarian failure. By using timed measurements of the replaced steroids to ensure attainment of premenopausal levels of the deficient hormones, the adequacy of an individual's therapy can be monitored. This method of hormone replacement has evolved over more than 10 years. It is inexpensive, gives good relief of symptoms, and is well tolerated. The endometrium is protected, and uterine bleeding is extremely infrequent. Patient compliance is excellent, and the positive effects of this hormone treatment encourage most patients to want long-term therapy. A general paradigm for HRT using natural sex steroids is given in Table 3. PMID- 7475495 TI - Abnormal uterine bleeding. AB - Physicians who care for female patients cannot avoid the frequent complaint of abnormal uterine bleeding. Knowledge of the disorders that cause this problem can prevent serious consequences in many patients and improve the quality of life for many others. The availability of noninvasive and minimally invasive diagnostic studies and minimally invasive surgical treatment has revolutionized management of abnormal uterine bleeding. Similar to any other disorder, the extent to which a physician manages abnormal uterine bleeding depends on his or her own level of comfort. When limitations of either diagnostic or therapeutic capability are encountered, consultation and referral should be used to the best interest of patients. PMID- 7475496 TI - Contraception. A clinical review for the internist. AB - Effective use of contraception allows couples to control their fertility and, in certain instances, is associated with important noncontraceptive benefits. Surgical sterilization offers safe, effective, and permanent contraception for women and men. Low-dose OCPs offer safe, effective contraception for most women able and willing to take pills consistently. In addition, their use confers a variety of important noncontraceptive benefits. The availability of long-acting progestin injectable and implantable methods has made highly effective and convenient birth control available for women who are not good pill takers as well as for those for whom contraceptive doses of estrogen are contraindicated. Barrier methods are readily available and provide protection against STDs; unfortunately, erratic use is all too often associated with inadequate contraceptive efficacy. IUDs offer convenient, highly effective contraception for appropriate candidates. PMID- 7475497 TI - Chronic pelvic pain. AB - The basic approach to the patient with chronic pelvic pain as practiced at the University of Tennessee, Memphis, has been described here. Using this approach allows the practicing physician to evaluate and manage realistically or refer appropriately most patients with chronic pelvic pain. The key to management of patients who have chronic pelvic pain is to use any and all available diagnostic and therapeutic modalities to identify the source(s) and direct therapy. By having at least a working knowledge of the various diagnostic and therapeutic modalities available to the patient, the individual clinician can better aid the patient in her understanding of chronic pelvic pain, generally, and her own pelvic pain, specifically. PMID- 7475498 TI - The abnormal Papanicolaou smear. AB - Despite its limitations, the Pap smear is a widely accepted, powerful screening technique for cellular abnormalities of the cervix. Abnormal Pap smears are found in 1% to 5% of the screened population, depending on the population characteristics. It is therefore important for all clinicians caring for women to be familiar with the principles of Pap smear screening, its strengths and weaknesses, the terminology used to interpret results, and the options for appropriate response to an abnormal smear. High-grade lesions require colposcopic examination and treatment; low-grade lesions require colposcopic examination and either treatment or close follow-up; atypical smears require some response, either repeated cytology or further investigation; glandular lesions require colposcopic examination and investigation of the endocervix. A tremendous amount of research is going on to assess the safest, most cost-effective approach to the detection, evaluation, and management of cervical epithelial lesions. It can be expected, therefore, that these recommendations will change over the next several years. All physicians providing health care for women should keep abreast of these changes. PMID- 7475500 TI - A clinician's guide to the premenstrual syndrome. AB - Many women have menstrual symptoms, but relatively few have severe PMS. PMS is a well-defined premenstrual cluster of predominantly affective symptoms that disrupt a woman's daily functioning. PMS is diagnosed with prospective charting of symptoms and should be differentiated from nondisruptive menstrual symptoms, major affective disorders, and other common medical and gynecologic conditions. Most women with PMS can be helped. The serotonin reuptake inhibitors are becoming the first line of therapy for PMS because they are effective, easily tolerated, and free of major side effects. There is also evidence supporting the role of other antidepressants, anxiolytics, and GnRH agonists in the treatment of PMS. Although increasing control of one's life, promoting a healthy diet, the avoidance of salt and caffeine, vitamin supplementation, and exercise have not been proved as effective treatment for PMS, they should be promoted for their obvious general health benefits. No one treatment fits the heterogeneous PMS population. A trial of medication should be continued for two or three menstrual cycles with appropriate dose adjustments. If relief is not sufficient, other agents or other treatments should be initiated. PMID- 7475499 TI - Breast disease. AB - Breast diseases are a common aspect of primary care practice. Common benign and malignant breast diseases are reviewed. The evaluation of common breast problems is presented and current recommendations for diagnostic and screening mammography are discussed. PMID- 7475501 TI - Pelvic organ prolapse and stress urinary incontinence. AB - Pelvic organ prolapse presents a wide array of distressing symptoms to the female patient. Stress urinary incontinence is often considered a normal phenomenon of aging and tolerated for years before seeking medical attention. A detailed history outlining specific complaints and a targeted pelvic examination can usually elucidate the anatomic problems responsible for producing symptoms. Nonsurgical therapy can be initiated, often with great improvement in symptoms. If the results are not satisfactory, the patient can be referred for further evaluation and possible surgical intervention. PMID- 7475502 TI - The female pelvic mass. Diagnosis and management. AB - When a pelvic mass is encountered in a female patient, there are several tools available to the physician as diagnostic aids. The patient's age, history, and physical examination; diagnostic imaging studies; and laboratory tests can contribute valuable information to the diagnosis and management of the case. It is essential for today's physician to become familiar with these tools and to be able to direct management appropriately. PMID- 7475503 TI - Conflicts and resolutions. PMID- 7475504 TI - Oligonucleotide diversity and drug discovery. PMID- 7475505 TI - The medical significance of physiological cell death. PMID- 7475506 TI - Molecular structure and tissue distribution of ryanodine receptors calcium channels. PMID- 7475507 TI - Selective inhibitors of monoamine oxidase (MAO-A and MAO-B) as probes of its catalytic site and mechanism. PMID- 7475508 TI - Electrophoretic evaluation of the mobility of drugs suitable for iontophoresis. AB - Iontophoresis is the process of increasing the penetration of drugs for therapeutic purposes using DC electricity. The most important factor for iontophoresis is the formation of drug ions in solution. On this basis, the penetration of any drug applied by iontophoresis might be maximized at an optimal pH. Seven medications were investigated: acyclovir, methylprednisolone sodium succinate, metronidazole, dexamethasone sodium phosphate, minoxidil, lidocaine hydrochloride, and adenine arabinoside monophosphate. Using techniques of paper electrophoresis, we evaluated the influence of pH on the mobility (mu) of the drugs in phosphate buffers at pH 5, 7, and 9. The mu-values were calculated after detecting the drugs on paper with ultraviolet light. The results indicate that the seven drugs had higher mu-values usually at one, but sometimes at two, of the pH values studied. This study suggests that a variation in pH might alter the maximum ionization, and therefore, the optimal mobility for these seven drugs, as well as other drugs, suited to iontophoresis. PMID- 7475509 TI - Comparative studies of Ca N-acetylhomotaurinate and Ca N-acetyltaurinate. I. Effects on the ionic transfer through the isolated human amnion. AB - The effects of two taurine and homotaurine derivatives (Ca N-acetyltaurinate: ATACa and Ca N-acetylhomotaurinate: AOTACa) were studied on the ionic transfer through a membranous model: the human isolated amniotic membrane. The ionic transfer was evaluated by measurements of the various components (cellular and paracellular) of the total conductance across the membrane. AOTACa influenced all the components of the total ionic conductance without flux ratio modifications and the general action scheme was a biphasic effect: decrease conductance with low concentrations and increase conductance with high concentrations. This effect might be important in the case of lower or higher supplementation. ATACa only interfered with some conductance components and this action was predominantly monophasic (decreased or increased conductance). This study indicates differential actions between two closely related molecules on a membranous model. PMID- 7475510 TI - Microwave-induced lethal heat stress: effects of phentolamine, prazosin and metoprolol. AB - Previous studies have shown that adrenergic antagonists can alter physiological responses to microwave-induced heating. In the present experiments, 33 ketamine anesthetized Sprague-Dawley rats were exposed to 2450-MHz microwaves at an average power density of 60 mW/sq cm (whole-body average specific absorption rate of approximately 14 W/kg) until death occurred. The effects of intraarterial injections of the nonselective alpha-adrenoceptor antagonist phentolamine (10 mg/kg body weight), the alpha 1-adrenoceptor antagonist prazosin (5 mg/kg), and the beta 1-adrenoceptor antagonist metoprolol (2 mg/kg) on physiological responses (including changes in body temperature, heart rate, blood pressure, and respiratory rate) were examined. A group of saline-injected animals was included for comparison. There were no significant differences in lethal temperatures, survival times, or rates of temperature change among the different groups. In all groups, heart rate increased continuously during exposure; mean arterial blood pressure increased until colonic temperature reached 41-41.5 degrees C, and then decreased. These heart rate and blood pressure changes were similar to those that occur during environmental heat stress. Despite differences in absolute values of heart rate and mean arterial blood pressure among groups, there were no significant differences in changes from baseline levels among groups. From these results, taken together with previous findings in our laboratory, it may be hypothesized that selective alpha 1 blockade (without concurrent alpha 2 blockade) and beta 2 blockade (with or without concurrent beta 1 blockade) are effective in altering physiological responses to microwave exposure. PMID- 7475511 TI - Scintigraphic imaging with 123I-metaiodobenzylguanidine as a marker of cardiac adrenergic neurotransmission in the rat. AB - This study was performed to determine if 123I-metaiodobenzylguanidine (123I-MIBG) could be used to create scintigraphic images of the rat heart and to determine if this method could be used to estimate cardiac norepinephrine turnover in the intact animal. Male F344 rats were injected i.v. with 200-350 muCi of 123I-MIBG. Beginning at 5 min after the injection, scintigraphic images were made every 30 min for 5 h. The half-life of 123I-MIBG retention within the heart during the 5 h of the experiment was determined by plotting the log of the counts for each image vs. time. The half-life of cardiac 123I-MIBG was 3.44 +/- 0.17 h with a rate constant of 0.21 +/- 0.01 (hr-1). In a separate experiment, methacholine, a parasympathomimetic agonist, was given after the injection of 123I-MIBG and increased 123I-MIBG efflux. Cardiac 123I-MIBG uptake was reduced by 65% in rats sympathetically denervated with 6 hydroxy dopamine indicating that 123I-MIBG is accumulating for the most part in adrenergic nerve terminals. We conclude that scintigraphic imaging with 123I-MIBG may be a useful procedure for estimating cardiac adrenergic nervous system activity in the intact animal. PMID- 7475512 TI - Diuretic activity of N'-disubstituted morpholinoguanidine analogs of U-37883A in rats and dogs. AB - U-37883A is a K+ sparing diuretic which selectively blocks openers of vascular ATP-sensitive K channels. Many N'-disubstituted morpholinoguanidine (N'-DMG) analogs of U-37883A were synthesized and tested for diuretic activity. In conscious rats, 10-100 mg/kg orally of the most active N'-DMGs increased urine volume (V) and Na+ excretion by up to 4-fold with little kaliuresis. The N'-DMGs U-37997A and U-38658A were less potent than standard diuretics, but did not induce the K+ loss seen with hydrochlorothiazide and furosemide or the K+ retention of amiloride and triamterene. In conscious dogs, 10 mg/kg i.v. of the N'-DMGs U-40389A and U-52090 increased V and Na+ excretion by over 7-fold with little kaliuresis. Despite their attractive diuresis, all of the N'-DMGs had narrow margins of safety. Reflecting their direct myocardial depressant action, in isolated rat hearts, bolus intracoronary U-37883A, U-18177A, and U-38658A (0.25-10 mumol) severely reduced the rate (-10 to -100%) and force (-9 to -100%) of contraction. These studies characterize the eukalemic diuretic activity of N' DMG analogs of U-37883A, and demonstrate the marked cardiac depression characteristic of the morpholinoguanidine diuretic series. PMID- 7475513 TI - Use of the pithed rat model to determine the relative potencies of 5-HT2A/2C receptor antagonists following acute intravenous and oral administration. AB - We investigated the usefulness of the pithed rat model to determine the relative potencies of 5-HT2A/2C receptor antagonists following acute intravenous and oral administration in inhibiting 5-HT-induced pressor responses. The 5-HT2A/2C receptor antagonists, pirenperone, ketanserin, and ritanserin, all dose dependently inhibited 5-HT-induced pressor responses when administered either intravenously 5 min prior to commencing 5-HT injections or acutely via the oral route 30 min before pithing. All three compounds were found to be of comparable potency in inhibiting 5-HT-induced pressor responses when given i.v. (range of ID50s 3.7-27.5 micrograms/kg), whereas substantial differences became apparent after oral administration (range of ID50s 13.0-214.0 micrograms/kg), as reflected also by the p.o./i.v. ID50 ratios for the drugs. In conclusion, inhibition of 5 HT-induced pressor responses in the pithed rat preparation provides a reproducible and robust procedure in which comparisons can be made of relative potencies of 5-HT2A/2C receptor antagonists, not only by the intravenous route, but also following acute oral administration. PMID- 7475514 TI - Failure of aspirin and diltiazem to prevent the formation of acute coronary thrombi in dogs. AB - The aim of the present study was to determine the efficacy of aspirin and diltiazem in preventing the formation of coronary thrombi in dogs. Canine coronary thrombi were produced by inserting a small catheter filled with collagen powder into the endothelial-injured, partially occluded left anterior descending coronary artery. Neither aspirin (bolus of 30 mg/kg, followed by 100 mg/kg/h by infusion), nor diltiazem (0.1 mg/kg, followed by 0.3 mg/kg/h by infusion) prevented the formation of coronary thrombi. The mortality in aspirin group was significantly higher than that in control and diltiazem groups. These results indicate that aspirin and diltiazem do not inhibit thrombus formation in the canine model of coronary thrombosis. PMID- 7475515 TI - Vascular effects of the 21-aminosteroid tirilazad mesylate: protection against oxidant stress. AB - The vascular effects of tirilazad mesylate (U-74006F), a 21-aminosteroid antioxidant under development for the treatment of acute CNS trauma and ischemia, have been investigated using isolated rings of rabbit aorta. Endothelium dependent relaxation was measured in rings contracted with 0.25 microM phenylephrine. Acetylcholine produced a dose-dependent relaxation that was slightly attenuated by the addition of U-74006F (0.1-10.0 microM). At a concentration of 10.0 microM, U-74006F shifted the EC50 for acetylcholine relaxation from 60 +/- 10 to 150 +/- 20 nM and reduced the maximum response by 20%. U-74006F (0.1-10.0 microM), did not reduce relaxation to the endothelium independent vasodilator nitroglycerin nor did it affect the tone of either resting or phenylephrine contracted rings. ACh relaxation was abolished by a 40 min treatment with the superoxide generating system xanthine oxidase (XO; 0.1 U/ml) plus xanthine (0.4 mM). Relaxation to nitroglycerin was not impaired by XO, nor were the phenylephrine-induced contractions. U-74006F, at doses of 0.05-10 microM, protected against XO mediated damage to endothelium-dependent relaxation. These results demonstrate that tirilazad mesylate can protect endothelial function from damage by reactive oxygen species. Preservation of endothelial function might represent an important component of the activity of tirilazad mesylate in vivo. PMID- 7475517 TI - [The oxygen-nurses are not optimally utilized]. PMID- 7475516 TI - [Quality of death--is it time for an open debate?]. PMID- 7475519 TI - [Increased salaries don't signify increased quality of health care]. PMID- 7475518 TI - [We don't receive sufficient information about calcium blockers and other drugs]. PMID- 7475521 TI - [Tether our sacred cows and more people can receive health care]. PMID- 7475520 TI - [On subarachnoidal hemorrhage, one more time]. PMID- 7475522 TI - [To manage chronic pain. Rehabilitation is a possible therapeutic alternative]. PMID- 7475523 TI - [Supposedly, calcium channel blockers cause infarction. Then the Bayer company took over the press conference]. PMID- 7475524 TI - [Pain rehabilitation is a pedagogic task. Chronic pain dealt with in a model for behavior-therapy]. PMID- 7475525 TI - [Successful diagnosis of tuberculosis. PCR determination of tuberculosis on formalin-fixed preparations]. PMID- 7475526 TI - [Lessons learned from the Ranealv accident. Role distribution important during crisis intervention]. PMID- 7475527 TI - [Upper airway obstruction. An unusual and life-threatening complication of Waran therapy]. PMID- 7475528 TI - [Dental braces against sleep apnea syndromes]. PMID- 7475529 TI - [Obstructive sleep apnea syndromes. Current status surveyed on a "state of the art" conference]. PMID- 7475530 TI - [Cellular telephones may interfere with cardiac stimulators. Yuppie telephones and alarms are hazardous for patients with pacemakers]. PMID- 7475531 TI - [The American Medical Association is in the middle of a political fight in health care]. PMID- 7475532 TI - [A general practitioner's dilemma--an odyssey directed by changing winds of politics]. PMID- 7475533 TI - [Irresponsible physicians!]. PMID- 7475534 TI - [Sectorized psychiatry is developable and cost-effective]. PMID- 7475535 TI - [Disturbed gastric function in certain patients with diabetes. Metabolic imbalance or autonomic neuropathy my be the cause]. PMID- 7475537 TI - [10 years of experiences with HIV and AIDS care. A report from a dermato venereological clinic]. PMID- 7475536 TI - [Gastric emptying is important for the patients with diabetes]. PMID- 7475538 TI - [Simple and painless for the patient with stomach ulcer. Blow and reveal the bacteria!]. PMID- 7475540 TI - [High protein intake in children increases the risk of obesity]. PMID- 7475539 TI - [Persons with multiple problems represent a heavy clientele. Psychiatric districts in Stockholm are carrying different load]. PMID- 7475541 TI - [The man behind the syndrome: Brown-Sequard. Experimental research absorbed his interest]. PMID- 7475542 TI - [Symptom--diagnosis or a signal? A colleague without own identity "reprimands" the body]. PMID- 7475543 TI - [Correct assessment of symptoms--an everyday task with a lot of "silent knowledge"]. PMID- 7475544 TI - [Training of ethical problem-solving. Support for terminal care personnel]. PMID- 7475545 TI - [Personal instruction in a new model of education]. PMID- 7475546 TI - Gaucher's disease variant characterised by progressive calcification of heart valves and unique genotype. AB - Gaucher's disease is the most prevalent sphingolipid storage disease, characterised by substantial genetic and phenotypic variability. Cardiac manifestations are rare. We report 12 Arab Gaucher's disease patients, 2-20 years of age, who presented with oculomotor apraxia but only slight classic signs of the disease. All but the youngest had calcifications of the aortic or mitral valves or both. All these patients were homozygous for the rare point mutation D409H (1342C). Valvular calcifications increased with age and showed progression during 2 years of follow-up. Two of the oldest patients underwent aortic valve replacement, and one sibling had died suddenly at age 16, before this study, Corneal opacities were another common feature. The potentially fatal course of this Gaucher's disease variant, and the availability of a reliable PCR-based method for heterozygote detection, mean that population screening and genetic counselling in the geographic area at risk are important. Affected individuals should be closely monitored by echocardiography to gauge the need for valve replacement. The potential of enzyme replacement to prevent these cardiac complications cannot be ascertained at present, because of the high cost of therapy. PMID- 7475547 TI - Microwaves for menorrhagia: a new fast technique for endometrial ablation. AB - Current ablative techniques for treating menorrhagia are unsatisfactory. We tried microwave energy, delivered by an 8 mm diameter probe in the uterine cavity, to treat 23 patients with functional menorrhagia. The mean treatment time was 2 min 12 s (range 50-310 s). 6 months after treatment the success rate is 83%, 13 patients (57%) are amenorrhoeic, and six (26%) are experiencing light menstruation. Three initial failures have been successfully retreated. The technique is safe, and is easier and quicker to perform than current alternatives. PMID- 7475549 TI - Possible mechanism involving T-lymphocyte response to non-structural protein 3 in viral clearance in acute hepatitis C virus infection. AB - In acute hepatitis C virus (HCV) infection only 20-50% of patients spontaneously clear the virus. To characterise the immune reaction during that phase we studied the response of peripheral blood mononuclear cells (PBMC) to the recombinant HCV proteins core, non-structural protein 3 (NS3), NS4, and NS5 in 14 patients with acute hepatitis C. All eight patients with self-limited disease compared with two of six with evolving chronic infection showed an NS3- specific PBMC response (p = 0.015). Of 65 patients with established chronic hepatitis C, five showed a PBMC response to NS3. NS3-specific CD4 T-cell clones from patients with self-limited infection predominantly produced interferon-gamma and may thus support cytotoxic effector mechanisms important for viral clearance. PMID- 7475550 TI - De-novo mutation and sporadic presentation of acute intermittent porphyria. AB - Acute intermittent porphyria (AIP) is a low-penetrant autosomal dominant disorder characterised by life-threatening neurovisceral attacks, often precipitated by drugs. Prognosis is improved by presymptomatic diagnosis and counselling. We found that 29 of 103 (28%) unrelated patients presented as sporadic cases. One patient had a de-novo mutation. The same mutation has arisen de novo at least three times in 12% of British AIP families. About 3% (95% CI 1-8%) of all AIP index cases may have de-novo mutations. These findings emphasise the high frequency of sporadic presentation of AIP and indicate the limitations of presymptomatic diagnosis as an aid to management. PMID- 7475548 TI - Extrinsic-pathway activation in cancer with high factor VIIa and tissue factor. AB - Thromboembolic complications are common in patients with malignant disease. We studied the activation of coagulation in 106 patients with solid tumours and 72 healthy volunteers by measuring plasma levels of tissue factor, factor VIIa, factor XIIa, thrombin-antithrombin complex, and prothrombin fragments 1 + 2. Tissue factor was 67% higher in cancer patients (median 582 vs 349 pg/mL, p = 0.0006) and factor VIIa was 46% higher (100 vs 69 mU/mL, p = 0.0002), indicating extrinsic pathway activation. Modest activation of the intrinsic pathway (elevated factor XIIa) was seen only in patients with advanced disease or those receiving chemotherapy. Excess thrombin generation was manifested by elevations in thrombin-antithrombin complex and prothrombin fragments 1 + 2. Tissue factor pathway is clearly implicated in the hypercoagulable state of cancer. PMID- 7475551 TI - Cancer progression and p53. AB - In a complex organism, somatic cells are under intermittent selection pressure for the emergence of mutants that can survive environmental insults and that can grow autonomously despite adverse conditions. Repeated rounds of mutation, selection, and proliferation may lead to cancer. The organism prevents malignant transformation by assuring accurate DNA repair before cell division, by forcing the death of cells with excessive DNA damage, and by placing limits on the replicative lifespans of most somatic cells. The p53 gene is a "guardian of the genome"--it regulates multiple components of the DNA damage control response and promotes cellular senescence. Disabling mutations and deletions of p53 occur in 50% of human tumours. p53-deficient cancers are often unstable, aggressive, and resistant to therapy. PMID- 7475552 TI - Evolution of complex disasters. PMID- 7475554 TI - A global witch-hunt. PMID- 7475553 TI - Is an elevated concentration of acinar cytosolic free ionised calcium the trigger for acute pancreatitis? AB - The pathogenesis of acute pancreatitis is poorly understood, despite well recognised precipitating factors. Current evidence suggests that the earliest abnormalities of acute pancreatitis arise within acinar cells, but the key intracellular trigger has yet to be identified. Within the pancreas, physiological concentrations of secretagogues bind to G-protein-linked cell surface receptors on acinar cells, evoking short, oscillatory spikes of acinar cytosolic-free ionised calcium ([Ca2+]i), an ubiquitous intracellular messenger. Specific effects within acinar cells include initiation of enzyme release through the phosphorylation cascades of stimulus-secretion coupling. Low resting levels of [Ca2+]i are restored by Ca(2+)-ATPase, which pumps calcium into the endoplasmic reticulum and out of the cell. If high concentrations of [Ca2+]i persist, toxicity results, intracellular signalling is disrupted, and cell damage occurs. Sustained elevations in acinar [Ca2+]i result from exposure to high concentrations of secretagogues, high doses of which also induce acute pancreatitis. Similarly, sustained elevations of [Ca2+]i may result from ductal hypertension, alcohol, hypoxia, hypercalcaemia, hyperlipidaemia, viral infection, and various drugs--all factors known to precipitate acute pancreatitis. We suggest that these factors precipitate acute pancreatitis by causing either excessive release of acinar [Ca2+]i, or damage to the integrity of mechanisms that restore low resting levels of [Ca2+]i, and that the consequent calcium toxicity is the key trigger in the pathogenesis of acute pancreatitis. PMID- 7475555 TI - Work stress can cause political strain. PMID- 7475556 TI - Anti-androgens in treatment of prostate cancer. PMID- 7475557 TI - Anti-androgens in treatment of prostate cancer. PMID- 7475558 TI - Anti-androgens in treatment of prostate cancer. PMID- 7475559 TI - Hypertension with moclobemide. PMID- 7475560 TI - Local anaesthetic on Filshie clips. PMID- 7475561 TI - Local anaesthetic on Filshie clips. PMID- 7475562 TI - Local anaesthetic on Filshie clips. PMID- 7475563 TI - Local anaesthetic on Filshie clips. PMID- 7475564 TI - Pirenzepine for clozapine-induced hypersalivation. PMID- 7475565 TI - Streptococcus pneumoniae resistant to penicillin and tetracycline associated with HIV seropositivity. PMID- 7475566 TI - Auditory hallucinations and perception of external speech. PMID- 7475567 TI - Auditory hallucinations and perception of external speech. PMID- 7475568 TI - Auditory hallucinations and perception of external speech. PMID- 7475569 TI - Severity of disease in cystic fibrosis. PMID- 7475570 TI - Flawed epidemiology of birth defects? PMID- 7475571 TI - Changing the definition of perinatal mortality. PMID- 7475573 TI - Polyauthoritis giftosa. PMID- 7475572 TI - Dangers of overviews of health care. PMID- 7475574 TI - UK NHS research and development system. PMID- 7475575 TI - Impact of clinical practice guidelines. PMID- 7475576 TI - Familial Alzheimer's disease associated with S182 codon 286 mutation. PMID- 7475577 TI - Effect of PCBs on spermatogenesis. PMID- 7475578 TI - Concerns about generalisation of premature ACAS recommendations for carotid endarterectomy. PMID- 7475579 TI - Muscle sounds rediscovered. PMID- 7475580 TI - HLA association with autoimmune disease: restricted binding or T-cell selection? PMID- 7475581 TI - Tuberculosis control policy in Nepal. PMID- 7475582 TI - Tuberculosis control policy in Nepal. PMID- 7475583 TI - Seat-belt syncope. PMID- 7475584 TI - Terminal restlessness and nicotine withdrawal. PMID- 7475585 TI - In defence of the FDA. PMID- 7475586 TI - Genotype/phenotype correlations in Gaucher's disease. PMID- 7475587 TI - Intraocular device for cytomegalovirus infection. PMID- 7475588 TI - Caring and sharing. PMID- 7475589 TI - Alpha-interferon in chronic myelogenous leukaemia. PMID- 7475590 TI - Peppermint oil and colon spasm. PMID- 7475591 TI - Determinants of progression to cirrhosis or fibrosis in pure alcoholic fatty liver. AB - "Pure" alcoholic fatty liver has been widely assumed to be "benign" with very low risk of progression to cirrhosis. Studies thus far have included either patients with coexisting recognised precursor lesions of cirrhosis or have been restricted to short-term histological follow-up. We have followed 88 patients, first seen between 1978 and 1985, with a histological diagnosis of pure alcoholic fatty liver and no evidence of fibrosis or alcoholic hepatitis, for a median of 10.5 years, to determine any factors predictive of disease progression. Of the 88, at follow-up nine had developed cirrhosis and a further seven fibrosis. Eight of nine patients with cirrhosis had continuing alcohol consumption of more than 40 units per week at follow-up; in the other patients, consumption was unknown. Independent histological predictors of progression on index biopsy were: presence of mixed macro/microvesicular fat, and presence of giant mitochondria. We can no longer regard alcoholic fatty liver as benign. In the presence of continuing high alcohol consumption the above histological features identified those at high risk (47-61%) of disease progression. Therefore, patients with these features should be counselled intensively regarding their alcohol consumption. PMID- 7475592 TI - Effectiveness of joint consultation sessions of general practitioners and orthopaedic surgeons for locomotor-system disorders. AB - Joint consultation sessions between general practitioners (GPs) and specialists to examine patients for whom decisions about referral are difficult are thought to be helpful, but their effects have not been evaluated. In a randomised, controlled trial we studied the effects of joint sessions of GPs and orthopaedic surgeons on referral and intervention rates. During 1.5 years, 12 GPs (in groups of three) held monthly joint consultation sessions with four participating orthopaedic surgeons: patients were seen by one orthopaedic surgeon in the presence of three GPs. Patients were included in the trial if the GP was uncertain about the diagnostic or therapeutic management and if referral was considered; and excluded if referral was urgently necessary or if there was some other clear indication for referral. By a randomised consent design, patients were assigned to joint consultation sessions (n = 144) or a usual-care control group (n = 128). A year later the patients were examined by an independent orthopaedic surgeon. There were significantly fewer referrals (51/144 [35%] vs 87/128 [68%], p < 0.01) and diagnostic actions in the intervention group than in the control group, without negative effects on health or functional status. More patients in the intervention group were symptom-free at 1 year (35% vs 24%, p < 0.05). Joint consultation sessions of GPs and orthopaedic surgeons within the framework of general practice resulted in more efficient care, with better targeted examination, treatment, and referrals. PMID- 7475594 TI - Turning to primary care. PMID- 7475593 TI - Breast and ovarian cancer incidence after infertility and in vitro fertilisation. AB - Concern has been expressed that exposure to fertility drugs might be associated with a risk of ovarian cancer. We have examined the incidence of breast and ovarian cancer in a cohort of 10,358 women referred for in-vitro fertilisation (IVF) treatment in Victoria, Australia, between 1978 and 1992. The "exposed" group (n = 5564) had had ovarian stimulation to induce multiple folliculogenesis and the "unexposed" group (n = 4794) had been referred for IVF but were untreated or had had "natural cycle" treatment without ovarian stimulation. Duration of follow-up ranged from 1 to 15 years. Cases of cancer were determined by record linkage with data from population-based cancer registries. 34 cases of invasive breast cancer and 6 of invasive ovarian cancer were observed. A comparison with the expected numbers, derived by applying age-standardised general population rates to the cohort gave standardised incidence ratios (SIR) for breast cancer of 0.89 (95% CI 0.55-1.46) in the exposed group and 0.98 (0.62-1.56) in the unexposed group, and for ovarian cancer SIRs were 1.70 (0.55-5.27) and 1.62 (0.52 5.02), respectively. Rates of all cancers were not significantly different from general population rates. The relative risk (RR) of cancer, adjusted for age and infertility type, was, in the treated group compared with the untreated group, 1.11 (95% CI 0.56-2.20) for breast cancer and 1.45 (0.28-7.55) for ovarian cancer. The risk of body of uterus cancer was increased in the exposed and unexposed groups combined (SIR 2.84 [1.18-6.81]). Women with unexplained infertility, independent of IVF exposure, had significantly increased risks of ovarian cancer (RR = 19.19 [2.23-165.0]) and body of uterus cancer (RR = 6.34 [1.06-38.0]) compared with women with known causes of infertility. This relatively short-term follow-up suggests that ovarian stimulation with IVF is not associated with an increased risk of breast cancer. Although there was no significantly increased risk of ovarian cancer after ovarian stimulation with IVF the small number of cases limits the conclusions that can be drawn. Longer-term follow-up of large cohorts of women who have been in IVF programmes will be necessary. PMID- 7475595 TI - Have expert panel guidelines kept pace with new concepts in hypertension? PMID- 7475596 TI - Intravenous thrombolysis in refractory unstable angina pectoris. PMID- 7475597 TI - Psychosocial care and breast cancer. PMID- 7475598 TI - The dilemma of laxative abuse. PMID- 7475599 TI - Hazards of misguided ethics committees. PMID- 7475600 TI - Sex mismatch as a risk factor for chronic rejection of liver allografts. AB - Chronic irreversible rejection is a major cause of graft loss and retransplantation after orthotopic liver allotransplantation. To identify risk factors we retrospectively analysed 423 adult consecutive primary liver allograft recipients. The endpoint of the study was graft failure due to chronic rejection leading either to retransplantation or death. Chronic rejection developed in 22 (5.2%) patients. Pretransplant diagnosis of primary biliary cirrhosis or autoimmune hepatitis, recipient age less than 30 years, 1 or more episodes of acute cellular rejection, and transplantation of an organ from cytomegalovirus (CMV). IgG positive donor to an IgG negative recipient were identified as risk factors for chronic rejection. Transplantation of a liver from a male donor into a female recipient was also associated with an increased probability of chronic rejection. By logistic regression analysis, the probability of chronic rejection was predicted by: sex and cytomegalovirus match of donor and recipient, the presence of acute rejection, recipient age, transplantation for autoimmune hepatitis or primary biliary cirrhosis, and recipients receiving no azathioprine during the third month after transplantation. Sensitisation to antigens expressed by bile-duct epithelium as in primary biliary cirrhosis or exposure to donor bile duct minor histocompatibility antigens, such as the male sex related H-Y antigen, may provide an explanation. More selective allocation of donor organs may allow a reduction in the incidence of ductopaenic rejection and graft loss. PMID- 7475601 TI - Influence of pregnancy on the course of primary chronic glomerulonephritis. AB - According to some nephrologists, pregnancy has damaging effects on renal function in primary glomerulonephritis, but the evidence is conflicting. We evaluated the effect of pregnancy on the occurrence of end-stage renal failure (ESRF) in 360 patients with various histological forms of primary glomerulonephritis but with normal renal function (serum creatinine < or = 0.11 mmol/L) at presentation. In actuarial analyses, overall ESRF-free survival did not significantly differ between women who became pregnant after clinical onset of renal disease (n = 171) and those who did not conceive (n = 189). Furthermore, in a case-control study pregnancy did not emerge as a risk factor for progression to ESRF (odds ratio 1.15 [95% CI 0.61-2.18]), whereas the type of glomerulonephritis and hypertension were major determinants. We conclude that pregnancy does not affect the course of renal disease in patients who have normal renal function at conception. PMID- 7475602 TI - Electrical stimulation of sacral spinal nerves for treatment of faecal incontinence. AB - Functional deficits of the striated anal sphincteric muscles without any apparent gross defect often result in a lack of ability to postpone defaecation by intention or in faecal incontinence in response to increased intra-abdominal or intra-rectal pressure. We applied electrostimulation to the sacral spinal nerves to increase function of the striated muscles of the anal sphincter. Of three patients followed for 6 months, two gained full continence and one improved from gross incontinence to minor soiling. Closure pressure of the anal canal increased in all. Preliminary data indicate that anal closure pressure increases with the duration of stimulation. Continuous stimulation of sacral spinal nerves can help some patients with faecal incontinence. It may be possible to promote continence with intermittent stimulation. PMID- 7475603 TI - Systemic haemodynamics and oxygenation during haemodilution in children. AB - Transfusion of homologous blood should be avoided when possible, and one technique that diminishes perioperative requirement for donor blood is haemodilution. In children its effects on systemic haemodynamics and systemic oxygenation have not been reported. Six children aged 4-12 yr were anaesthetised for major surgery and blood was withdrawn to reduce packed cell volume to 25%. Cardiac index increased from 3.1 (SD 0.5) L min-1 m-2 at baseline to 4.4 (0.5) L min-1 m-2 at the end of surgery, when packed cell volume was 16 (1)%; this change, compensating for the decline in oxygen carrying capacity, was associated with a fall in systemic vascular resistance and a rise in stroke volume. Oxygen extraction from haemoglobin rose from 0.22 (0.05) to 0.33 (0.06). Perioperative blood loss was 40% of circulating blood volume; however, owing to reinfusion of autologous blood (and use of a cell saver in three patients), the haemoglobin one day after operation was only 19% lower than preoperatively (9.9 [1.5] vs 12.5 [2.5] g/dL). In this study, children seemed at least as able as adults to compensate for the effects of haemodilution, which allowed major surgery without transfusion of homologous blood. PMID- 7475604 TI - Presynaptic dopamine function in striatum of neuroleptic-naive schizophrenic patients. AB - Presynaptic dopamine function (6-[18F]-fluorodopa uptake) in the brains of seven neuroleptic-naive first-admission schizophrenic patients and eight healthy controls was studied with positron emission tomography. The fluorodopa influx constant (Ki) in putamen was higher in the patients than in controls (average mean: 0.0149 vs 0.0129, p = 0.034). The changes in caudate were smaller but significantly lateralised to the left caudate. There was one catatonic schizophrenic patient in our sample. This patient had lower striatal Ki than any control. Alterations in striatal presynaptic dopamine function may constitute a part of disrupted neural circuits that predispose to schizophrenic psychosis. PMID- 7475605 TI - Detection of the GBV-C hepatitis virus genome in serum from patients with fulminant hepatitis of unknown aetiology. AB - Hepatitis viruses A, B, C, D, and E have not accounted for all cases of hepatitis, hence "non A-E" agent(s) might be implicated. A set of new viruses (GBV-A, -B, and -C) whose genomes have been sequenced, are being investigated as possible causes of non A-E hepatitis. We investigated six cases of fulminant hepatitis of unknown aetiology for the presence of GBV-C genome in their serum, and three showed positive signals by semi-nested PCR using primers derived from the NS3/helicase region. Nucleotide sequence analyses confirmed these signals to be derived from a GBV-C sequence. The results suggest the importance of GBV-C in the aetiology of fulminant hepatitis. PMID- 7475606 TI - World distribution of factor V Leiden. AB - We have analysed 3380 chromosomes (1690 unrelated individuals) from twenty-four populations for the presence of factor V Leiden, an important risk factor in venous thromboembolism. The allele frequency in 618 Europeans was 4.4%, with the highest prevalence among Greeks (7%). It was 0.6% in Asia Minor. Factor V Leiden was not found in any of 1600 chromosomes from Africa, Southeast Asia, Australasia, and the Americas. This distribution may partly explain the rarity of thromboembolic disease in these populations. The high prevalence in Europeans suggests that screening for this mutation should be considered in some circumstances. PMID- 7475607 TI - Application of artificial neural networks to clinical medicine. PMID- 7475608 TI - General practitioner fundholding: weighing the evidence. AB - A new development in the British National Health Service is fundholding, whereby certain general practitioners are given budgets from which they purchase services for patients. Our knowledge about fundholding is rudimentary. Many important questions remain unanswered. These include: the impact of fundholding on the efficiency of overall resource utilisation; its effect on strategic planning; its effect on equity; its impact on the role of the doctor and the doctor-patient relationship; and its consequences for practice organisation and the culture of primary care. Even the scant results we do have must be interpreted cautiously. Fundholding was introduced during a period of great turbulence for general practice. Its strategic function was uncertain and its impact has been confounded by the effects of a series of earlier and concurrent policy changes. Few reliable conclusions about fundholding, either positive or negative, can be drawn from existing research. PMID- 7475609 TI - Humanity and the medical humanities. PMID- 7475611 TI - Sporadic Creutzfeldt-Jakob disease in a 16-year-old in the UK. PMID- 7475610 TI - Labour faces health nightmares. PMID- 7475612 TI - Sporadic Creutzfeldt-Jakob disease in a 18-year-old in the UK. PMID- 7475613 TI - Hepatitis B reactivation after lamivudine. PMID- 7475615 TI - Impact of improved treatment of sexually transmitted disease on HIV infection. PMID- 7475614 TI - Impact of improved treatment of sexually transmitted disease on HIV infection. PMID- 7475616 TI - Impact of improved treatment of sexually transmitted disease on HIV infection. PMID- 7475617 TI - Impact of improved treatment of sexually transmitted disease on HIV infection. PMID- 7475618 TI - Impact of improved treatment of sexually transmitted disease on HIV infection. PMID- 7475619 TI - Effect of angiotensinogen gene T235 variant on the development of diabetic complications in type II diabetes mellitus. PMID- 7475620 TI - Trading trust for blood money. PMID- 7475621 TI - History of umbilical cord blood transplantation. PMID- 7475622 TI - Quality of chloroquine preparations marketed in Dar es Salaam, Tanzania. PMID- 7475623 TI - Trading trust for blood money. PMID- 7475624 TI - Genetic basis of adult-onset spinal muscular atrophy. PMID- 7475625 TI - The challenge of tuberculosis. PMID- 7475626 TI - The challenge of tuberculosis. PMID- 7475627 TI - The challenge of tuberculosis. PMID- 7475628 TI - The challenge of tuberculosis. PMID- 7475629 TI - The challenge of tuberculosis. PMID- 7475630 TI - Clinical trial compensation. PMID- 7475632 TI - Euthanasia and the ICU. PMID- 7475631 TI - Dealing with duffers. PMID- 7475633 TI - Hepatitis E in Brazil. PMID- 7475634 TI - Use of long-acting contraceptives. PMID- 7475635 TI - Maternal haemoglobin and the secretion of chorionic gonadotropin in early human pregnancy. PMID- 7475636 TI - Treatment of localised scleroderma by UVA1 phototherapy. PMID- 7475637 TI - Deaths and injuries caused by landmines. PMID- 7475639 TI - Deaths and injuries caused by landmines. PMID- 7475638 TI - Enteritis, eosinophilia, and Enterobius vermicularis. PMID- 7475640 TI - Unknown multiresistant gram-negative bacterium causing meningitis in HIV-positive patient in Africa. PMID- 7475641 TI - Need for liver transplantation. PMID- 7475642 TI - Need for liver transplantation. PMID- 7475643 TI - Protrusion of retention wire from Encor-Dec passive fixation atrial pacing lead. PMID- 7475644 TI - Nef vaccination against HIV disease. PMID- 7475645 TI - Evidence-based medicine. PMID- 7475646 TI - Evidence-based medicine. PMID- 7475647 TI - Evidence-based medicine. PMID- 7475648 TI - Evidence-based medicine and Kurt Godel. PMID- 7475649 TI - Evidence-based medicine. PMID- 7475650 TI - Evidence-based medicine. PMID- 7475651 TI - Maintaining confidentiality. PMID- 7475652 TI - Angioplasty versus bypass surgery. PMID- 7475653 TI - Nervous about artificial neural networks? PMID- 7475654 TI - Microsatellites: heroes and villains. PMID- 7475655 TI - Is prostate cancer worth diagnosing? PMID- 7475656 TI - First-year results of CABRI (Coronary Angioplasty versus Bypass Revascularisation Investigation). CABRI Trial Participants. AB - The Coronary Angioplasty versus Bypass Revascularisation Investigation (CABRI) is a multinational, multicentre randomised trial comparing the strategies of revascularisation by CABG (coronary artery bypass grafting) and PTCA (percutaneous transluminal coronary angioplasty) in patients with symptomatic multivessel coronary disease. 1054 patients (820 men and 234 women) were recruited from 26 European cardiac centres. The average age was 60 years and 62% presented with angina of class 3 or greater. 513 patients were randomised to CABG and 541 to PTCA, and 93% and 96%, respectively, of those randomised underwent the allocated procedure. This first report presents data analysed by intention to treat and documents all deaths, major cardiac events, and the symptom status of the patients 1 year after randomisation. After 1 year of follow-up, 14 (2.7%) of those randomised to CABG and 21 (3.9%) of those randomised to PTCA had died. The PTCA group's relative risk (RR) of death was 1.42 (95% CI 0.73-2.76). Patients randomised to PTCA required significantly more reinterventions; only 66.4% reached 1 year with a single revascularisation procedure compared with 93.5% of patients randomised to CABG (RR = 5.23 [3.90-7.03], p < 0.001). The patients in the PTCA group took significantly more medication at 1 year (RR = 1.30 [1.18 1.43], p < 0.001). They were also more likely to have clinically significant angina (RR = 1.54 [1.09-2.16], p = 0.012); this association was present in both sexes but was significant only in females. CABRI is the largest trial of CABG versus PTCA to be reported so far. Its findings are consistent with previous studies, and add to the weight of information that clinicians need to discuss with patients when options for the management of severe angina are under consideration. PMID- 7475657 TI - Meta-analysis of randomised trials comparing coronary angioplasty with bypass surgery. AB - A patient with severe angina will often be eligible for either angioplasty (PTCA) or bypass surgery (CABG). Results from eight published randomised trials have been combined in a collaborative meta-analysis of 3371 patients (1661 CABG, 1710 PTCA) with a mean follow-up of 2.7 years. The total deaths in the CABG and PTCA groups were 73 and 79, respectively, with a relative risk (RR) of 1.08 (95% CI 0.79-1.50). The combined endpoint of cardiac death and non-fatal myocardial infarction occurred in 169 PTCA patients and 154 CABG patients (RR 1.10 [0.89 1.37]). Amongst patients randomised to PTCA 17.8% required additional CABG within a year, while in subsequent years the need for additional CABG was around 2% per annum. The rate of additional non-randomised interventions (PTCA and/or CABG) in the first year of follow-up was 33.7% and 3.3% in patients randomised to PTCA and CABG, respectively. The prevalence of angina after one year was considerably higher in the PTCA group (RR 1.56 [1.30-1.88]) but at 3 years this difference had attenuated (RR 1.22 [0.99-1.54]). Overall there was substantial similarity in outcome across the trials. Separate analyses for the 732 single-vessel and 2639 multivessel disease patients were largely compatible, though the rates of mortality, additional intervention, and prevalent angina were slightly lower in single vessel disease. The combined evidence comparing PTCA with CABG shows no difference in prognosis between these two initial revascularisation strategies. However, the treatments differ markedly in the subsequent requirement for additional revascularisation procedures and in the relief of angina. These results will influence the choice of revascularisation procedure in future patients with angina. PMID- 7475658 TI - Randomised placebo-controlled trial of primaquine for prophylaxis of falciparum and vivax malaria. AB - Drug resistance has made malaria prevention difficult and the new agents are too expensive for widespread use. Primaquine, an established drug for treatment, is potentially useful for prevention. Malaria prophylaxis with primaquine was evaluated in Irian Jaya during one year in Javanese men who were not deficient in glucose-6-phosphate dehydrogenase (G-6-PD). 126 volunteers were randomised to receive 0.5 mg/kg primaquine base or placebo daily (double-blinded), or 300 mg chloroquine base weekly (open). The protective efficacy of primaquine relative to placebo was 94.5% (95% confidence interval 57-99) for Plasmodium falciparum and 90.4% (95% CI 58-98) for P vivax. Attack rates for either parasite did not differ significantly between the chloroquine and placebo groups. Incidence density of physical complaints not associated with parasitaemia was low (17-18 complaints/person-year) and was about the same in all groups except for cough, which was increased in the primaquine group. Complete blood counts were normal and no evidence of hepatic or renal dysfunction was found with primaquine. However, at 50 weeks the primaquine group had a mean methaemoglobin of 5.8% (range 1.4-13%), which declined by half within 7 days of ending prophylaxis. When used daily for one year by men with normal G-6-PD activity, primaquine was well tolerated and effective for prevention of malaria. PMID- 7475659 TI - Slowing of wound healing by psychological stress. AB - There is evidence that psychological stress adversely affects the immune system. We have investigated the effects of such stress, caused by caring for a relative with Alzheimer's disease, on wound healing. We studied 13 women caring for demented relatives (mean age 62.3 [SE 2.3] years) and 13 controls matched for age (60.4 [2.8] years) and family income. All subjects underwent a 3.5 mm punch biopsy wound. Healing was assessed by photography of the wound and the response to hydrogen peroxide (healing was defined as no foaming). Wound healing took significantly longer in caregivers than in controls (48.7 [2.9] vs 39.3 [3.0] days, p < 0.05). Peripheral-blood leucocytes from caregivers produced significantly less interleukin-1 beta mRNA in response to lipopolysaccharide stimulation than did controls' cells. Stress-related defects in wound repair could have important clinical implications, for instance for recovery from surgery. PMID- 7475660 TI - Multiple introductions of HIV-1 subtype E into the western hemisphere. AB - There are nine recognised genetic subtypes of HIV-1, and the epidemic in Southeast Asia is largely due to subtype E. We have investigated HIV-1 viral subtypes in 11 Uruguayan military personnel, six with infection acquired during a United Nations deployment to Cambodia and five with infection acquired in South America. We found subtype E in five of the six infections acquired in Southeast Asia, and subtype B in all five of the domestically acquired cases. These findings document multiple introductions of HIV-1 subtype E into the western hemisphere and mean that the genetic diversity of the global HIV-1 pandemic must be considered in strategies for epidemic control. PMID- 7475661 TI - Detection of diverse HIV-1 genetic subtypes in the USA. AB - Of the nine genetic subtypes of HIV-1 that exist world wide, subtype B predominates in North America and Europe. Thus, most knowledge about HIV-1 and most vaccine development efforts are based on subtype B viruses. We document here the detection of HIV-1 subtypes A, D, and E in five US servicemen who acquired these non-subtype-B infections during overseas deployments. The dispersal of diverse HIV-1 subtypes into regions of the world with previously restricted genetic diversity may have important implications for the epidemiology of the epidemic and for the design and implementation of vaccine trials. PMID- 7475662 TI - Diagnostic use of microsatellite instability in hereditary non-polyposis colorectal cancer. AB - 50 families with a history of colorectal cancer were divided according to whether criteria for hereditary non-polyposis colorectal cancer (HNPCC) were fulfilled totally (A, n = 19) or partly (B, n = 31) and stratified by the demonstration that at least half the cancers tested per family were positive for DNA replication errors (RER+). Accepted clinical and pathological characteristics of HNPCC were found to cluster within 12 A/RER+ families in which the mean number of affected individuals per family was 10.1. Reliance upon clinical data alone may result in over-diagnosis of HNPCC, in small families who just meet the minimum criteria, whereas underdiagnosis is rare. The criteria could be refined by inclusion of RER status. PMID- 7475663 TI - Liquid ventilation in adults, children, and full-term neonates. AB - We evaluated the safety and efficacy of partial liquid ventilation in a series of 19 adults, children, and neonates who were in respiratory failure and on extracorporeal life support. During partial liquid ventilation, the alveolar arterial oxygen difference decreased from 590 (SE 25) to 471 (42) mm Hg (p = 0.0002) and static pulmonary compliance increased from 0.18 (0.04) to 0.29 (0.04) mL cm H2O-1 kg-1 (p = 0.0002). 11 patients (58%) survived. These preliminary data suggest that partial liquid ventilation can be safely used in patients with severe respiratory failure and may improve lung function. PMID- 7475664 TI - Artificial neural networks in pathology and medical laboratories. PMID- 7475665 TI - Proposed link between transmissible spongiform encephalopathies of man and animals. AB - A link between scrapie and Creutzfeldt-Jakob disease (CJD) is likely to exist. Based on old observations on scrapie, new experiments on bovine spongiform encephalopathy, and modern reviews on CJD, my proposal fits general rules of virus transmission. PMID- 7475666 TI - The next pandemic influenza virus? PMID- 7475667 TI - William Cooke of Exeter: apothecary, surgeon, and grave-robber. PMID- 7475668 TI - Republican revolution moves along. PMID- 7475669 TI - Disclosure of medical records, in Canada. PMID- 7475670 TI - Emergency percutaneous carotid stenting during stroke. PMID- 7475671 TI - Thyroid cancer in children in Belarus: ascertainment bias? PMID- 7475672 TI - Melanoma and sun exposure. PMID- 7475673 TI - Pooled plasma derivatives and Creutzfeldt-Jakob disease. PMID- 7475674 TI - Melanoma and sun exposure. PMID- 7475675 TI - Melanoma and sun exposure. PMID- 7475676 TI - Tackling methicillin-resistant Staphylococcus aureus. PMID- 7475677 TI - Risperidone-induced extrapyramidal reactions. PMID- 7475678 TI - Sodium bicarbonate does not correct respiratory acidosis. PMID- 7475679 TI - Alcohol sclerotherapy for benign solitary solid cold thyroid nodules. PMID- 7475680 TI - Folate prophylaxis in pregnancy. PMID- 7475681 TI - Gastroparesis with multiple sclerosis. PMID- 7475682 TI - Remission of epilepsy. The NGPS. National General Practice Study of Epilepsy. PMID- 7475683 TI - Combined spinal-epidural or standard epidural analgesia in labour. PMID- 7475684 TI - Melatonin for treatment of chronic refractory sarcoidosis. PMID- 7475685 TI - Crimean-Congo haemorrhagic fever in Oman. PMID- 7475688 TI - Forgotten imprisoned Syrian doctors. PMID- 7475687 TI - Potentiation of placebo analgesia by proglumide. PMID- 7475686 TI - Malabsorption of vitamin E in cystic fibrosis improved after ursodeoxycholic acid. PMID- 7475690 TI - Global burden of tropical diseases. PMID- 7475689 TI - Epidemic malaria in India's Thar desert. PMID- 7475691 TI - Ehrlichiosis in Belgium. PMID- 7475692 TI - Lichen planus associated with hepatitis C. PMID- 7475693 TI - Plague in Majunga, Madagascar. PMID- 7475694 TI - Ciprofloxacin in sweat and antibiotic resistance. The Copenhagen Study Group on Antibiotics in Sweat. PMID- 7475695 TI - Condensed expirate nitrite as a home marker for acute asthma. PMID- 7475696 TI - Management of paracetamol poisoning. PMID- 7475697 TI - Management of paracetamol poisoning. PMID- 7475698 TI - Management of paracetamol poisoning. PMID- 7475699 TI - Erroneous genetic risk assessment of Alport syndrome. PMID- 7475700 TI - Management of paracetamol poisoning. PMID- 7475701 TI - Screening of blood donors for human parvovirus B19. PMID- 7475702 TI - Clobazam for seizure prophylaxis during busulfan chemotherapy. PMID- 7475703 TI - Tuberculosis in prisons. PMID- 7475704 TI - Tuberculosis in prisons. PMID- 7475705 TI - HCV in Egypt in 1977. PMID- 7475706 TI - Delayed gastric emptying in bulimic patients. PMID- 7475707 TI - Sea-urchin injuries. PMID- 7475708 TI - Deglamorising cannabis. PMID- 7475709 TI - Pneumocystis carinii in Africa: an emerging pathogen? PMID- 7475710 TI - Who's to blame for asthma? PMID- 7475711 TI - Climate and disease in Colombia. PMID- 7475712 TI - HIV-1, the brain, and combination therapy. PMID- 7475713 TI - Exit trans fatty acids. PMID- 7475714 TI - Learning from stories--The Lancet's case reports. PMID- 7475715 TI - Fibrosing colonopathy in cystic fibrosis: results of a case-control study. AB - Fibrosing colonopathy was first described in cystic fibrosis (CF) children in 1994. We have done a nested case-control study to identify possible associations with this condition. A case ascertainment within the UK CF population to identify any cases that occurred between January, 1984, and April, 1994, found 14 cases, all under 14 years and confirmed by independent histopathological review. All had presented since April, 1993; 12 were boys and six had received some or all of their care in Liverpool. Each case was matched, by date of birth, with four controls from the UK CF Registry. Information was obtained about cases and controls from their case records and by a structured interview with the families. In the 12 months before surgery, there was an association between the occurrence of fibrosing colonopathy and use of high-strength pancreatic enzyme preparations. This association was dose related. Odds ratio per extra 1000 high-strength capsules was 1.45 (95% CI 1.14-1.84). For use of protease, the odds ratio per million extra units per kg was 1.55 (1.19-2.03). For usage of individual high strength products at any time during the 12 months before surgery some differences were observed; for Creon 25000 the odds ratio was 0.38 (0.10-1.42), for Nutrizym 22 43.4 (2.51-751), and for Pancrease HL 8.4 (1.95-36.1). These last two confidence intervals are extremely wide and compatible with these two products having the same odds ratios. Laxative use was independently predictive (odds ratio 2.42 [1.20-4.94]). We conclude that there is a dose-related association between high-strength pancreatic enzyme preparations and fibrosing colonopathy. PMID- 7475716 TI - Serum, breast milk, and infant antibody after maternal immunisation with pneumococcal vaccine. AB - Pneumococci are a leading cause of severe bacterial disease in infants and children world wide. A possible means of protecting infants in the first few months of life is immunisation of the mother during pregnancy. We prospectively assessed pneumococcal immunisation of pregnant women to determine the amount of pneumococcal antibody transmitted to the infants in serum and milk and the half life of the passively acquired antibody. Healthy pregnant women in Dhaka, Bangladesh, were randomised to receive pneumococcal or meningococcal vaccine with routine prenatal tetanus immunisation at 30-34 weeks of gestation. Serum and breast milk specimens from the mothers and sera from infants were collected up to 22 weeks of age and assayed for specific serum IgG, IgG1, and IgG2 and for milk IgA antibodies to pneumococcal serotypes 6B and 19F. 55 mothers and 56 infants were followed from birth to five months. Women who received pneumococcal vaccine had geometric mean antibody increases of 2.6 and 3.4 to types 6B and 19F, respectively. The mean infant/maternal antibody ratios were 0.56 and 0.59 (range 0.11-1.46) for these serotypes. Infant cord antibody titres correlated with maternal titres. Infant/maternal IgG ratios correlated with the interval between immunisation and birth and were higher for specific IgG1 than for IgG2. Infants of pneumococcal vaccine recipients had geometric mean antibody concentrations of 6.8 and 7.5 micrograms/mL to serotypes 6B and 19F in cord blood; in cord blood and in all subsequent serum specimens the concentrations were 2-3 fold higher than in control infants. The median half-life of passive antibody was about 35 days; at five months of age 63-71% of infants of pneumococcal vaccine recipients had antibody concentrations greater than 0.15 micrograms/mL. Breast milk IgA antibodies for pneumococcal serotype 19F, but not for type 6B, were significantly higher in vaccine recipients up to five months after delivery. If maternal pneumococcal polysaccharide antibodies do not interfere with active immunisation of the infant with new glycoprotein conjugate pneumococcal vaccines, passive active immunisation of infants can be a feasible strategy for developing regions. PMID- 7475717 TI - Pneumocystis carinii pneumonia in Zimbabwe. AB - Pneumocystis carinii pneumonia (PCP) is said to be rare in Africa, with reported rates of 0-22% in human-immunodeficiency-virus (HIV) infected individuals with respiratory symptoms. Over one year in a central hospital in southern Africa, 64 HIV-infected patients with acute diffuse pneumonia unresponsive to penicillin and sputum smear-negative for acid-fast bacilli underwent fibreoptic bronchoscopy. Bronchoalveolar lavage fluid was assessed for bacteria, fungi, Pneumocystis carinii, and mycobacteria. 21 patients (33%) had PCP and 24 (39%) had tuberculosis; 6 of these had both infections. 5 patients had Kaposi's sarcoma (KS) associated with PCP, tuberculosis, or another infection, in 1 patient KS was the only finding, and in 21 no pathogen was identified. A logistic regression model was used to assess clinical, radiographic, and arterial blood gas predictors of PCP and tuberculosis. Fine reticulonodular shadowing on the chest radiograph (nodular component < 1 mm) was the strongest independent predictor of PCP (odds ratio 8.5 [95% CI 6.1-10.9]). A respiratory rate of more than 40/min was the best clinical predictor of PCP (odds ratio 11.2 [95% CI 8.8-13.6]). Median CD4+ T cell count for all cases of PCP was 134/microL (range 5-355) and for tuberculosis without PCP 206/microL (range 61-787). In resource-limited countries, a regionally appropriate management algorithm is required. PMID- 7475718 TI - Linkage of high-affinity IgE receptor gene with bronchial hyperreactivity, even in absence of atopy. AB - Asthma is a manifestation of bronchial hyperreactivity (BHR) and forms part of the spectrum of atopic disease. Some pedigree studies of atopy have suggested linkage with the high-affinity IgE receptor (Fc epsilon RI beta) gene on chromosome 11q13, but others find no linkage. The molecular genetics of asthma and BHR have not been studied in the general population. We examined the genetic linkage of the Fc epsilon RI beta gene with clinical asthma and the underlying phenotypes of BHR (to methacholine) and atopy (defined by skinprick testing) in 123 affected sibling-pairs recruited from the general population. We found evidence of significant linkage of a highly polymorphic microsatellite marker in the fifth intron of the Fc epsilon RI beta gene to a diagnosis of asthma (18.0% excess of shared alleles, p = 0.002) and to BHR (21.7% excess of shared alleles, p = 0.001). Significant linkage was also observed in siblings sharing BHR when those with atopy were excluded (32.8% excess of shared alleles, p = 0.004). Atopy in the absence of BHR did not show significant linkage to the Fc epsilon RI beta gene (7.2% excess of shared alleles, p = 0.124). These findings suggest that mutations in the Fc epsilon RI beta gene or a closely linked gene influence the BHR underlying asthma, even in the absence of atopy. PMID- 7475719 TI - Gut inflammation in children with cystic fibrosis on high-dose enzyme supplements. AB - We used a whole-gut perfusion technique to study subclinical gut inflammation in children with cystic fibrosis (18 elective tests, three lavages to treat distal intestinal obstruction syndrome); and in 12 control children with constipation or pre-colonoscopy. We assayed for haemoglobin, IgG, albumin, alpha-1-antitrypsin, granulocyte elastase, interleukin-1 beta (IL-1 beta) and IL-8 concentrations in whole-gut lavage fluid. Results for two children with distal intestinal obstruction syndrome, the only children in the series taking Nutrizym 22, were strikingly abnormal. This new test has revealed subclinical gut mucosal inflammation in a minority of CF children, for which distal intestinal obstruction syndrome, Nutrizym 22 treatment, or both, may be risk factors. PMID- 7475720 TI - Telomerase activity in human brain tumours. AB - Malignant gliomas are invasive into surrounding brain and are refractory to therapy. Telomerase stabilises telomere length and may immortalise cells to allow unlimited proliferation. Our analysis of telomerase activity in 90 human gliomas showed that 19 of 19 oligodendrogliomas and 38 of 51 glioblastoma multiformes have detectable telomerase activity. The absence of telomerase activity in anaplastic astrocytomas (2/20 positive) and in one-quarter (13/51) of the glioblastomas suggests that these tumours follow different pathways of neoplastic progression. Thus we have found that a distinct subgroup of brain tumour consists of transformed yet pre-immortal cells. PMID- 7475721 TI - A sore eye and meningitis. PMID- 7475722 TI - 31-year-old woman with an enlarged tender liver. PMID- 7475724 TI - Information lives; medical television dies. PMID- 7475723 TI - Antimicrobial therapy in expectant management of preterm premature rupture of the membranes. AB - We review the impact of antimicrobial treatment on maternal and fetal outcome during expectant management of preterm premature rupture of the membranes. Relevant studies were retrieved from Medline (1966 to August, 1994) with the search term fetal-membrane-premature-rupture and antibiotics or antimicrobial, Excerpta Medica (1972 to August, 1994) with the search term premature fetus, membrane rupture, and antibiotic or antimicrobial therapy, and the Cochrane database of systemic reviews with the criterion antibiotics and prelabour rupture of membranes. We also obtained unpublished data from a randomised clinical trial of ceftizoxime versus placebo. The selected studies were randomised controlled trials of systemic antimicrobial therapy for prolongation of gestation in non labouring women after preterm premature rupture of the membranes. Data extraction was done by a single reviewer. Studies were evaluated for post-randomisation exclusion and other confounding variables that might introduce analytical bias. Analysis was done with SAS statistical software by a blinded investigator. Antimicrobial therapy after preterm premature rupture of the membranes is associated with a reduced number of women delivering within 1 week (62 vs 76%; OR 0.51, 95% CI 0.41-0.68), and reduced diagnosis of maternal morbidity including chorioamnionitis (12 vs 23%; 0.45, 0.33-0.60) and postpartum infection (8 vs 12%; 0.63, 0.41-0.97). Fetal morbidity, including confirmed sepsis (5 vs 9%; 0.57, 0.36-0.88), pneumonia (1 vs 3%; 0.32, 0.11-0.96), and intraventricular haemorrhage (9 vs 14%; 0.65, 0.45-0.92) were less often diagnosed after antimicrobial therapy. Separate analysis of the six placebo-controlled trials revealed similar or improved odds of pregnancy prolongation, chorioamnionitis, neonatal sepsis, postpartum infection, positive infant blood cultures, and pneumonia. Antimicrobial therapy, when used in the expectant management of preterm premature rupture of the membranes is associated with prolongation of pregnancy and a reduction in the diagnosis of maternal and infant morbidity. Further study should be directed towards determination of optimal antimicrobial therapy, increasing pregnancy prolongation, and enhancement of corticosteroid therapy for induction of pulmonary maturity after preterm premature rupture of the membranes. PMID- 7475725 TI - Politics of hard and soft drugs. PMID- 7475726 TI - Thalidomide effective for AIDS-related oral ulcers. PMID- 7475727 TI - Update on HIV transmission and pathogenesis. PMID- 7475728 TI - Endometrial changes in patients on tamoxifen. PMID- 7475729 TI - Endometrial changes in patients on tamoxifen. PMID- 7475730 TI - Kwashiorkor, depression, and attachment disorders. PMID- 7475731 TI - Safety of topical dinitrochlorobenzene. PMID- 7475732 TI - Mefloquine in malaria prophylaxis: low efficacy or low compliance? PMID- 7475733 TI - Malaria, anaemia, and HIV-1 transmission in central Africa. PMID- 7475734 TI - Intra-arterial iloprost for limb ischaemia. PMID- 7475735 TI - Magnesium for eclampsia. PMID- 7475736 TI - Childhood poisonings from ingestion of cigarettes. PMID- 7475737 TI - Transmission of hepatitis C virus by monoclonal-purified viral-attenuated factor VIII concentrate. PMID- 7475738 TI - Transmission of hepatitis C virus by monoclonal-purified viral-attenuated factor VIII concentrate. PMID- 7475739 TI - Aerosol inhalation before reading age. PMID- 7475740 TI - Idiopathic hypothalamic dysfunction: a paraneoplastic syndrome? PMID- 7475741 TI - Toxic interaction of venlafaxine and isocarboxazide. PMID- 7475742 TI - Melatonin secretion and coronary heart disease. PMID- 7475743 TI - Evidence-based medicine. PMID- 7475744 TI - Evidence-based medicine. PMID- 7475745 TI - What would Archie Cochrane have said? PMID- 7475746 TI - Impact factor as the best operational measure of medical journals. PMID- 7475747 TI - Impact factor as the best operational measure of medical journals. PMID- 7475748 TI - Euthanasia. PMID- 7475749 TI - Ciprofloxacin-resistant Salmonella typhi in the UK. PMID- 7475750 TI - Dopamine receptors in schizophrenia. PMID- 7475751 TI - Migration and AIDS. PMID- 7475752 TI - Migration and AIDS. PMID- 7475753 TI - HIV-associated diarrhoea and wasting. PMID- 7475754 TI - HIV and peacekeeping operations in Cambodia. PMID- 7475755 TI - Neuroprotective mechanism of bromocriptine. PMID- 7475756 TI - Streptococcus suis meningitis in an Italian blood donor. PMID- 7475757 TI - Alcohol consumption. PMID- 7475758 TI - Hoarse cry: a sign of dehydration. PMID- 7475759 TI - Anabolic steroid abuse and renal-cell carcinoma. PMID- 7475760 TI - Plasma exchange in Kawasaki disease. PMID- 7475761 TI - Magnesium sulphate increases plasma noradrenaline and neuropeptide-Y-like immunoreactivity. PMID- 7475762 TI - Intestinal perforation after ingestion of a blister-wrapped tablet. PMID- 7475763 TI - Inquiring into sex. PMID- 7475764 TI - Tacrolimus for prevention of liver allograft rejection: clinical trials and tribulations. PMID- 7475765 TI - What can a worm teach us about autoimmunity? PMID- 7475766 TI - Herodotus and the multidisciplinary clinic. PMID- 7475767 TI - Does atrial fibrillation confer a hypercoagulable state? PMID- 7475768 TI - Prospective screening of dyspeptic patients by Helicobacter pylori serology. AB - Helicobacter pylori infection is associated with 95% of duodenal ulcers and more than 80% of gastric ulcers. Several reports have indicated that screening for H pylori may avoid subsequent endoscopic examination. We screened 183 dyspeptic patients, aged under 45, by taking a history of sinister symptoms and regular use of non-steroidal anti-inflammatory drugs (NSAIDs), together with serological testing for H pylori. Endoscopy was performed on 113 patients, of whom 90 (49%) were seropositive, 14 (8%) had sinister symptoms, and 9 (5%) had used NSAIDs regularly. In 34 (19%) patients we detected peptic ulceration. The remaining 70 (38%) patients who were H pylori seronegative, had no sinister symptoms, and had not taken NSAIDs (screen-negative), did not undergo endoscopy but were returned to their primary care physician for treatment of symptoms. At subsequent reassessment (of the non-endoscoped group), symptom severity (p = 0.002), interference with life events (p = 0.01), and medication (p = 0.0002) were all significantly lower in the 6 months after screening than in the 6 month period before screening. Only three screen-negative patients were re-referred after screening but their endoscopic findings were normal. Thus, 67 (36%) endoscopies were avoided. When the non-endoscoped screen-negative patients were compared with a cohort of endoscoped screen-negative patients, the groups did not differ in terms of symptom severity (odds ratio 1.12, 95% CI 0.53-2.35, p = 0.77) or interference with life events (0.82, 0.38-1.76, p-0.61). However, medication use was significantly less (0.37, 0.17-0.80, p = 0.01) in those who did not have an endoscopy. Our study indicates that colonisation screening based on H pylori serology, a history of sinister symptoms, or a history of NSAID use was worthwhile in dyspeptic patients. We avoided 37% of endoscopies and reduced drug usage without disadvantaging those not endoscoped. PMID- 7475769 TI - Reduction of cellular rejection and increase in longer-term survival after heart transplantation after HLA-DR matching. AB - HLA matching in cardiac transplants is perceived as being logistically difficult. We studied 1135 consecutive primary cardiac allografts between 1980 and 1994 to assess the effect of HLA mismatching on long-term graft survival and cellular rejection episodes within 3 months of transplantation. We found a significant association between HLA-DR mismatching and the number of episodes of rejection (no mismatch 0.80 [SE 0.13], one mismatch 1.22 [0.06], two mismatches 1.42 [0.06], p < 0.05). We found a similar correlation between the total number of biopsy specimens showing evidence of cellular rejection and HLA-DR mismatch. The time between operation and the first rejection episode shortened with increasing HLA-DR mismatch (no mismatch 85.5 [37.3] days, one mismatch 43.1 [8.1], two mismatches 24.1 [2.9], p < 0.05). Furthermore, the proportion of patients with no evidence of rejection correlated with HLA-DR incompatibility. A significant association between improved graft survival and HLA-DR mismatching was found over 1, 5, and 10 years after transplantation (no mismatch 1 year 92%, 5 years 83%, 10 years 76%, one mismatch 1 year 81%, 5 years 73%, 10 years 59%, and two mismatches 78% 1 year, 5 years 70%, and 10 years 52%, p = 0.02). Increased efforts to prospectively HLA match patients has resulted in 25% of patients transplanted between January and May 1995 (n = 13/52) receiving grafts matched for HLA-DR. HLA matching reduces the frequency and severity of acute cardiac allograft rejection and improves graft survival for up to 10 years. Our preliminary results suggest that it is possible to use HLA matching prospectively for our selection of recipients. PMID- 7475770 TI - Return of function after spinal cord implantation of avulsed spinal nerve roots. AB - Avulsion of nerve roots from the spinal cord is widely regarded as an untreatable injury. However, a series of experiments in animals has shown that, if continuity is restored between spinal cord and ventral roots, axons from spinal motor neurons can regrow into the peripheral nerves with recovery of motor function. These observations were applied in the treatment of a man with avulsion of the 6th cervical (C6) to 1st thoracic roots due to brachial plexus injury. Two ventral roots were implanted into the spinal cord through slits in the pia mater, C6 directly and C7 via sural nerve grafts. Voluntary activity in proximal arm muscles was detected electromyographically after nine months and clinically after one year. After three years the patient had voluntary activity (with some co contraction) in the deltoid, biceps, and triceps muscles. To determine whether the improvement was due to spontaneous recovery from C5, the C5 root was blocked pharmacologically, and the results indicated that the repaired roots were contributing substantially to motor function. Repair of spinal nerve roots deserves further exploration in management of brachial plexus injury. PMID- 7475771 TI - Pertussis in adults: frequency of transmission after household exposure. AB - Although pertussis in adults is well documented, opinions differ about incidence of adult disease and about the role of adults as reservoirs of infection. We made use of a prospective household contact study of an acellular pertussis vaccine to collect data about pertussis in adults. All members of families with an index case of pertussis were monitored for respiratory symptoms, and pertussis was confirmed by laboratory tests. In 122 households, 104 children (85%) and 18 adults (15%) were the source of pertussis. These households consisted of 265 adults (aged 19-83 years), in 84 of whom (31%) pertussis was confirmed. Of these 84, 81% had respiratory symptoms for 21 days or more. The spread of pertussis was independent of whether a child (74/104) or an adult (14/18) was the index case. Most adult index cases had no pertussis recall (odds ratio 11.8). The overall attack rate in adult contacts was 0.267 and was independent of the social status and the size of the family and of a pertussis recall, although it differed significantly between women and men (p < 0.05). Erythromycin treatment of the index case reduced the attack rate significantly (p < 0.05). Patients whose first pertussis episode dated back more than 20 years had similar symptoms and attack rates to patients without a recall. We conclude that adults are often involved in the spread of pertussis, and that they can be susceptible to reinfection 20 years after a first pertussis episode. PMID- 7475772 TI - Time for a review of admission to medical school? AB - Appropriate selection of medical students is a fundamental prerequisite if medical schools are to produce competent and caring doctors. The selection criteria for entry to the medical degree course at the University of Newcastle, New South Wales, are unique in Australia. The purpose of this study was to identify admission criteria that may predict performance in the first postgraduate (intern) year. Performance ratings were obtained from the clinical supervisors of two graduating classes of Newcastle University medical students during their five terms in internship (first postgraduate year). At least one rating was obtained for 93% of interns. A subset analysis of interns with multiple ratings (57%) showed that combining previous study in both humanities and science before medical school entry was predictive of higher intern performance ratings. These interns were rated more favourably than those who had studied science alone. Moreover, students who had earlier studied both humanities and science were twice as likely to complete their medical degree as those who had studied science alone. Age, gender, admission interview results, written psychometric test scores, academic marks, and whether previous tertiary study had been undertaken prior to medical school entry were not predictive of intern performance ratings. Subject spread, including a background in humanities, is important for effective medical practice, at least in the immediate postgraduate period. Perhaps it is time to evaluate the admission criteria by which medical students are selected. PMID- 7475773 TI - Association between tumour angiogenesis and tumour cell shedding into effluent venous blood during breast cancer surgery. AB - Tumour angiogenesis is a powerful prognostic indicator in breast cancer. Shedding of tumour cells into the bloodstream is essential for haematogenous metastasis. The relation between cell shedding and angiogenesis in human cancer is not known. We studied vascular density and cell shedding in 16 women undergoing breast cancer surgery. Circulating cells were found in one patient before, in six during, and none after operation. Vascular density was related to detection of circulating tumour cells during surgery (correlation coefficient for first intraoperative sampling = 0.56, p = 0.024, 95% CI 0.08 to 0.83). Tumour cell shedding during surgery is related to vascular density; the relevance of this relation to the prognostic significance of vascular density in breast cancer merits further study. PMID- 7475774 TI - Observational study of effect of intrauterine transfusions on outcome of fetal hydrops after parvovirus B19 infection. AB - The role of intrauterine transfusion for fetal hydrops arising from maternal paravirus B19 infection is unclear. 66 cases of fetal hydrops arising from B19 infection were reported in England and Wales between June, 1992, and September, 1994. In 29 cases the fetus was dead at the time of the first abnormal ultrasound or a therapeutic abortion as performed; 12 of the 38 alive at the first abnormal scan received intrauterine transfusions and 3 of the 12 died. 26 did not receive intrauterine transfusions and 13 died. After adjustment for the severity of the hydrops as assessed by the ultrasound and for gestational age, the odds of death among those who received an intrauterine transfusion was significantly less than among those who did not (odds ratio 0.14, 95% CI 0.02-0.96). These findings suggest that intrauterine transfusion will benefit some fetuses with hydrops arising from parvovirus B19 infection. PMID- 7475775 TI - A diver unconscious after gastroenteritis. PMID- 7475776 TI - Variation in protection by BCG: implications of and for heterologous immunity. PMID- 7475778 TI - Retrospective on dark days at NCI. PMID- 7475777 TI - Randomised trialomania? The multicentre liver transplant trials of tacrolimus. PMID- 7475779 TI - Attenuated HIV-1 strains share gene deletions. PMID- 7475780 TI - FDA advised to license three anti-HIV agents. PMID- 7475781 TI - Netherlands revises HIV coordination efforts. PMID- 7475783 TI - Kaposi's sarcoma and KSHV. PMID- 7475782 TI - Kaposi's sarcoma and KSHV. PMID- 7475784 TI - Kaposi's sarcoma and KSHV. PMID- 7475785 TI - Kaposi's sarcoma and KSHV. PMID- 7475786 TI - Needlestick malaria. PMID- 7475787 TI - Fetal choroid plexus cysts. PMID- 7475788 TI - Fetal choroid plexus cysts. PMID- 7475789 TI - Vaginal lisuride for hyperprolactinaemia. PMID- 7475790 TI - Bacteraemia due to penicillin-resistant Streptococcus viridans in cancer patients, before and after prophylaxis with penicillin. PMID- 7475791 TI - Antibody response to hepatitis B vaccine among haemodialysis patients. PMID- 7475792 TI - Intravenous anti-D immunoglobulin for childhood acute immune thrombocytopenic purpura. PMID- 7475794 TI - Magnesium and eclampsia. PMID- 7475793 TI - BSE and farmworkers. PMID- 7475795 TI - S182 protein in Alzheimer's disease neuritic plaques. PMID- 7475796 TI - Blood pressure rise and ischaemic stroke. PMID- 7475797 TI - Antiquity of the Lyme-disease spirochaete in Europe. PMID- 7475798 TI - Therapeutic failure in group C streptococcal pharyngitis. PMID- 7475799 TI - Resigned to my fate. PMID- 7475800 TI - Necropsy as a research method. UK Cancer Family Study Group. PMID- 7475801 TI - Megatrials in myocardial infarction. PMID- 7475802 TI - Do refugees belong in camps? PMID- 7475803 TI - Interferon-alpha 1 for chronic myeloid leukaemia. PMID- 7475804 TI - Detection of parasite DNA in Chagas' heart disease. PMID- 7475805 TI - Is HIV the cause of AIDS? PMID- 7475806 TI - Long-term non-progression in HIV infection. PMID- 7475807 TI - Dual infection with HIV-1 Thai subtype B and E. PMID- 7475808 TI - Do we need pigmented lesion clinics? PMID- 7475809 TI - Naltrexone in alcohol dependence. PMID- 7475810 TI - Early antibiotic treatment in acute necrotising pancreatitis. PMID- 7475811 TI - Early antibiotic treatment in acute necrotising pancreatitis. PMID- 7475812 TI - Tonsillectomy: a family-transmissible surgical procedure. PMID- 7475813 TI - Coming clean about needle exchange. PMID- 7475814 TI - Statins prevent coronary heart disease. PMID- 7475815 TI - Breast cancer in female flight attendants. PMID- 7475816 TI - Prevention of cytomegalovirus disease in transplant recipients. PMID- 7475817 TI - Ouabain: a new steroid hormone? PMID- 7475818 TI - Preventing adhesions. PMID- 7475819 TI - Effect of GPi pallidotomy on motor function in Parkinson's disease. AB - The major motor disturbances in Parkinson's disease are thought to be caused by overactivity of the internal segment of the globus pallidus (GPi), in large part due to excessive drive from the subthalamic nucleus. The excessive inhibitory activity of GPi is thought to "brake' the motor thalamus and the cortical motor system to produce the slowness, rigidity, and poverty of movement characteristic of parkinsonian states. To test the hypothesis that direct reduction of Gpi activity can improve motor function, we studied the effect of GPi pallidotomy in 14 patients. The location of the GPi nucleus was confirmed by microelectrode recording before lesion creation. Standardised videotape recordings before and after operation were randomised and scored by a "blinded' evaluator. 6 months after surgery, total motor score in the "off" state had improved by 30% and the total akinesia score by 33%. The gait score in the "off" state improved by 15% and a composite postural instability and gait score by 23%. After surgery there was almost total elimination of drug-induced involuntary movements (dyskinesias), with a 92% reduction on the side contralateral to the pallidotomy. No patient had visual or corticospinal complications. In these patients GPi pallidotomy enhanced motor performance, reduced akinesia, improved gait, and eliminated the neural elements responsible for levodopa-induced dyskinesias. PMID- 7475820 TI - Apolipoprotein E allele epsilon 4, dementia, and cognitive decline in a population sample. AB - From clinically based series it has been proposed that, in homozygotes for the apolipoprotein E epsilon 4 (apoE epsilon 4) allele, Alzheimer's disease is almost inevitable by the age of 80. A population sample of persons aged 70 years and over was interviewed in 1990-91 to ascertain the presence of dementia or cognitive impairment. The sample was reinterviewed in 1994, when the apoE genotype was also determined. Prevalence data for the 638 persons who completed the second examination revealed a linear association between having an apoE epsilon 4 allele and both dementia and cognitive impairment (for heterozygotes, odds ratio for dementia 1.89, 95% confidence interval 1.04-3.44 and for homozygotes OR 3.58, 95% CI 1.08-11.82; both adjusted for age). However, even in subjects homozygous for epsilon 4 the estimated prevalence of dementia by age 90 was only about 50%. Persons with one or two epsilon 4 alleles were more likely to have a family history of dementia than those with none. This study confirms in a population sample that the epsilon 4 allele is a risk factor for dementia, but refutes the suggestion that homozygosity for the epsilon 4 allele is sufficient for the development of Alzheimer's disease: persons with either one or two epsilon 4 alleles may reach late old age without cognitive impairment. PMID- 7475822 TI - Prospective study of serum total homocysteine concentration and risk of stroke in middle-aged British men. AB - Moderate hyperhomocysteinaemia is common in the general population and has been linked with cardiovascular disease. However, there are no data from prospective, population-based studies. We examined the association between serum total homocysteine (tHcy) concentration and stroke in a nested case-control study within the British Regional Heart Study cohort. Between 1978 and 1980 serum was saved from 5661 men, aged 40-59 years, randomly selected from the population of one general practice in each of 18 towns in the UK. During follow-up to December, 1991, there were 141 incident cases of stroke among men with no history of stroke at screening. Serum tHcy was measured in 107 cases and 118 control men (matched for age-group and town, without a history of stroke at screening, who did not develop a stroke or myocardial infarction during follow-up). tHcy concentrations were significantly higher in cases than controls (geometric mean 13.7 [95% CI 12.7-14.8] vs 11.9 [11.3-12.6] mumol/L; p = 0.004). There was a graded increase in the relative risk of stroke in the second, third, and fourth quarters of the tHcy distribution (odds ratios 1.3, 1.9, 2.8; trend p = 0.005) relative to the first. Adjustment for age-group, town, social class, body-mass index, hypertensive status, cigarette smoking, forced expiratory volume, packed-cell volume, alcohol intake, diabetes, high-density-lipoprotein cholesterol, and serum creatinine did not attenuate the association. These findings suggest that tHcy is a strong and independent risk factor for stroke. PMID- 7475821 TI - Multicentre prospective controlled study of Barrett's oesophagus and colorectal adenomas. Groupe d'Etude de l'Oesophage de Barrett. AB - Previous studies have suggested that patients with Barrett's oesophagus may be at increased risk of colorectal neoplasia, though the association is disputed. In a multicentre prospective study we compared the prevalence of colorectal adenomas in patients with Barrett's oesophagus and controls. Barrett's oesophagus patients (n = 104) had histological confirmation of columnar epithelium extending more than 3 cm above the gastro-oesophageal junction. The 537 controls were patients with symptoms suggesting irritable bowel syndrome. No participant had a personal history of colonic neoplasm. Each patient underwent colonoscopy. Histologically proven adenomas were found in 26 Barrett's patients (25%) and 75 controls (14%). Three colorectal cancers were discovered in each group. The prevalence of adenomas was greater in the Barrett's oesophagus group than in the control group (p < 0.01) but the relation became non-significant after adjustment for age and sex and control for other known risk factors by a logistic regression model (odds ratio 1.4 [0.7-2.7]). The relative risk of adenoma was significantly higher in patients older than 59 than in younger patients (2.2 [1.3-3.5]) and in men than in women (3.4 [2.0-5.7]). Other factors contributing significantly to the risk of adenoma were a family history of colorectal cancer (2.3 [1.1-4.8]), rectal bleeding (2.1 [1.1-3.9]), previous colonic investigation (0.3 [0.1-0.7]), and complete as opposed to partial colonoscopy (6.4 [0.8-48.3]). We conclude that Barrett's oesophagus is not an independent risk factor for colorectal neoplasia and, therefore, is not, in itself an indication for colorectal screening. PMID- 7475823 TI - Vaccination with mutant ras peptides and induction of T-cell responsiveness in pancreatic carcinoma patients carrying the corresponding RAS mutation. AB - Mutations in codon 12 of K-RAS are frequently found in pancreatic adenocarcinomas. T-cell responses specific for individual RAS mutations can be elicited in vitro by stimulation of peripheral blood mononuclear cells with synthetic peptides. Mutant ras peptides are therefore a candidate vaccine for specific immunotherapy in pancreatic carcinoma patients. When vaccinated with a synthetic ras peptide representing the K-RAS mutation in their tumours, a transient ras-specific T-cell response was induced in two of five patients treated. The vaccination protocol involved multiple infusions of large amounts of peptide-pulsed antigen-presenting-cells obtained by leucapheresis. These results indicate that specific T-cell responses against mutations uniquely harboured in tumour cells can be induced in cancer patients by vaccination. PMID- 7475824 TI - Scintigraphic localisation of steroid injection site in plantar fasciitis. AB - Plantar fasciitis is a common cause of heel pain. We evaluated scintigraphic localisation of the inflammatory focus in 15 patients with this condition. Technetium-labelled bone scans precisely localised abnormal discrete areas of tracer uptake in 12 (80%) of patients in the medial and posterior aspect below the inferior surface of the calcaneum. Steroid injection at the inflammatory site abolished local tenderness and reduced pain in all 12. Our findings support an injection approach through the medial heel border posterior to the point of heel tenderness. Technetium scintigraphy may be a useful investigation to localise the steroid injection site in resistant cases of plantar fasciitis. PMID- 7475825 TI - Progressive blurring of vision in both eyes. PMID- 7475826 TI - Angiotensin receptor antagonists: focus on losartan. PMID- 7475827 TI - Pathophysiology and management of sickle cell pain crisis. Report of a Meeting of Physicians and Scientists, University of Texas Health Science Center at Houston, Texas. PMID- 7475828 TI - British health rationing becomes explicit. PMID- 7475829 TI - Safety of calcium antagonists. PMID- 7475830 TI - Normal serum gastrin concentration in gastrinoma. PMID- 7475831 TI - Risk of breast cancer after renal or cardiac transplantation. PMID- 7475832 TI - Risk of breast cancer after renal or cardiac transplantation. PMID- 7475833 TI - Human recombinant DNase for mucus plugging in status asthmaticus. PMID- 7475834 TI - Apolipoprotein E genotype and cerebral amyloid angiopathy. PMID- 7475835 TI - QT dispersion and ventricular fibrillation in acute myocardial infarction. PMID- 7475837 TI - Mortality and haemophilia. PMID- 7475836 TI - HIV-infected healthcare workers: results from a straw poll. PMID- 7475838 TI - HIV infection in Cuban women. PMID- 7475839 TI - Can type C hepatitis infection be complicated by malignant lymphoma? PMID- 7475840 TI - Singapore's haze and acute asthma in children. PMID- 7475841 TI - Hydroquinone and hepatitis. PMID- 7475842 TI - Hyperosmolar coma and lithium-induced diabetes insipidus. PMID- 7475843 TI - Hyperosmolar coma and lithium-induced diabetes insipidus. PMID- 7475844 TI - Hyperosmolar coma and lithium-induced diabetes insipidus. PMID- 7475845 TI - Hyperosmolar coma and lithium-induced diabetes insipidus. PMID- 7475846 TI - Oral contraceptives and venous thromboembolism. PMID- 7475847 TI - Oral contraceptives and venous thromboembolism. PMID- 7475848 TI - Oral contraceptives and venous thromboembolism. PMID- 7475849 TI - Cholera deaths in Goma. PMID- 7475850 TI - Dealing with duffers. PMID- 7475851 TI - Pros and cons of standardised mortality ratios. PMID- 7475852 TI - Haematological study of healthy volunteers 5 years after G-CSF. PMID- 7475853 TI - Is Hodgkin's disease a human counterpart of bacterially induced crown-gall tumours? PMID- 7475854 TI - Mutation of beta 3-adrenergic-receptor gene and response to treatment of obesity. PMID- 7475855 TI - Low-dose versus high-frequency regimens in Gaucher's disease. PMID- 7475856 TI - Occupational asthma induced by ethylene oxide. PMID- 7475857 TI - Scanning laser tomography in benign intracranial hypertension. PMID- 7475858 TI - Pitfalls of PCR: misdiagnosis of cerebral nocardia infection. PMID- 7475859 TI - Soil silicates and Kaposi's sarcoma in Sardinia. PMID- 7475861 TI - Lead contamination of candy: an example of crisis management in public health. PMID- 7475860 TI - Chagas' disease and dinitrochlorobenzene. PMID- 7475862 TI - In-vitro culture of testicular sperm. PMID- 7475863 TI - Of worms and red herrings. PMID- 7475865 TI - Restructuring the workforce in otolaryngology-head and neck surgery: it's time to play "what if"? PMID- 7475864 TI - Screening for familial hypercholesterolaemia in relatives. PMID- 7475866 TI - Transmastoid labyrinthectomy in older patients. AB - Transmastoid labyrinthectomy is an effective procedure for the control of episodic vertigo due to unilateral peripheral vestibular dysfunction. However, its application in older patients has been limited, primarily because of concerns that older patients may not compensate well postoperatively. Poor vestibular compensation results in constant disequilibrium, motion intolerance, and visual symptomatology. The purpose of this report is to review our results with transmastoid labyrinthectomy in patients over the age of 65. Complete records were available for 22 patients operated on between July 1984 and June 1994. Patient's records were reviewed for age, preoperative duration of symptoms, coexistent medical conditions, vertigo control, and postoperative disequilibrium. Advanced age need not be a contraindication to transmastoid labyrinthectomy. PMID- 7475867 TI - Minor salivary gland tumors of the palate: clinical and pathologic correlates of outcome. AB - Minor salivary gland tumors of the palate are rare and may pose a diagnostic and therapeutic dilemma for the head and neck surgeon. The authors reviewed their 46 years of experience with minor salivary gland tumors of the palate to determine the factors that influence outcome and their implications for treatment. Malignant tumors were seen in 116 patients (78%) and benign tumors were found in 33 patients (22%). Adenoid cystic carcinoma was the most common malignant tumor, occurring in 43 patients, and pleomorphic adenoma was the most common benign tumor, occurring in 30 patients. Univariate analysis on the malignant lesions showed that grade 3 tumor histology (P < .001), tumor size greater than 3 cm (P < .001), perineural invasion (P = .031), bone invasion (P = .012), positive surgical margins (P < .001), and positive initial but negative final margins (P = .004) were all associated with decreased survival. With multivariate analysis, tumor size, margin status, and grade were shown to be independently associated with decreased survival (P < .05). The recurrence rate at the primary site was significantly higher for adenoid cystic carcinoma than for other histologies (P = .0059). The 2-, 5-, and 10-year disease-specific survival rates for patients with malignant disease were 96%, 87%, and 80%, respectively. Wide surgical excision with adequate margins is essential for a favorable outcome in patients with malignant minor salivary gland tumors. Postoperative radiotherapy is reserved for patients with grade 3 tumor histology, large primary lesions, perineural invasion, bone invasion, cervical lymph node metastasis, and positive margins, although a clear-cut survival advantage has not been proven. Recurrence, especially regional and distant metastasis, portends an extremely poor prognosis. PMID- 7475868 TI - Modified transnasal endoscopic Lothrop procedure: further considerations. AB - The Lothrop procedure resects the medial frontal sinus floor, superior nasal septum, and intersinus septum to create a large frontonasal communication. However, the external approach often allowed medial collapse of soft tissue and stenosis of the nasofrontal communication. We describe a modified transnasal endoscopic Lothrop procedure in which drills are used for cases in which frontal recess exploration fails to relieve obstruction of the frontal sinus. The lateral bony walls are preserved, and medial collapse does not occur. The mucosa of the posterior table and posterior nasofrontal duct is preserved, and a single common frontal opening is created. We have found this approach to be safe and reliable. Fourteen patients have undergone this procedure without complication, achieving resolution or improvement of their symptoms and maintaining wide patency of the frontonasal opening. We recognize that long-term follow-up will be required but remain encouraged with our favorable results to date. PMID- 7475869 TI - Distribution of efferent cholinergic terminals and alpha-bungarotoxin binding to putative nicotinic acetylcholine receptors in the human vestibular end-organs. AB - Although acetylcholine (ACh) has been identified as the primary neurotransmitter of the efferent vestibular system in most animals studied, no direct evidence exists that ACh is the efferent neurotransmitter of the human vestibular system. Choline acetyltransferase immunohistochemistry (ChATi), acetylcholinesterase (AChE) histochemistry, and alpha-bungarotoxin binding were used in human vestibular end-organs to address this question. ChATi and AChE activity was found in numerous bouton-type terminals contacting the basal area of type II vestibular hair cells and the afferent chalices surrounding type I hair cells; alpha bungarotoxin binding suggested the presence of nicotinic acetylcholine receptors on type II vestibular hair cells and on the afferent chalices surrounding type I hair cells. This study provides evidence that the human efferent vestibular axons and terminals are cholinergic and that the receptors receiving this innervation may be nicotinic. PMID- 7475870 TI - Safety of thyroidectomy in residency: a review of 186 consecutive cases. AB - A retrospective review was performed to assess the surgical complications of thyroidectomy performed by otolaryngology residents under close supervision by faculty. One hundred eighty-six consecutive cases of thyroidectomy performed by the residents in the Department of Otolaryngology--Head and Neck Surgery at the University of Southern California were reviewed. Sixty-nine percent of thyroidectomies were performed for malignant or possible malignant diseases, and 31% were performed for benign conditions. Transient (less than 2 weeks in duration) hypocalcemia occurred in 26% of the patients; the vast majority of cases resolved within the first week. Five percent of the patients developed persistent hypocalcemia requiring prolonged replacement therapy. The incidence of unexpected postoperative permanent vocal cord paralysis was 0.9%. These complication rates are comparable to those in previously published large series on results of thyroidectomy. Thyroidectomy performed in an otolaryngology residency training program is a safe procedure when closely supervised by experienced faculty. PMID- 7475871 TI - Outcome analysis of Mohs surgery of the lip and chin: comparing secondary intention healing and surgery. AB - Wounds of the lip and chin resulting from microscopically controlled (Mohs) surgery are often repaired immediately. However, wounds allowed to heal by secondary intention have the advantage of optimal cancer surveillance, simplified wound care, and avoidance of the costs and potential complications associated with reconstructive procedures. Accurate prediction of the course of wound healing would allow a rational approach to selection of surgery or healing by secondary intention. The authors evaluated 105 patients with defects of the lip and chin after Mohs excision for cancer who healed by secondary intention. Forty six patients treated surgically were followed for comparison. The lip and chin were divided into subunits and the wound location, size, and depth were recorded. Patients were followed at intervals and a final determination regarding cosmesis was made after 6 or more months. It is concluded that the final cosmetic result can be confidently predicted on the basis of location by subunit, size, and depth of the wound. PMID- 7475872 TI - Cochlear implantation in the congenital malformed cochlea. AB - Cochlear implantation has proven beneficial for numerous children with profound congenital hearing impairments. Some congenitally deaf ears, however, may have anatomical malformations. This study reports the authors' recent experience with cochlear implants in patients with cogenital inner ear malformations. Since 1987, 10 patients with congenital abnormal inner ear structures have been implanted at the House Ear Clinic; 8 with abnormal cochleas and 2 with enlarged vestibular aqueducts. Nine patients received the Nucleus 22 device and 1 patient was implanted with the 3M/House device. Some modifications of the routine surgical approach were required in 5 patients. A cerebrospinal fluid (CSF) leak occurred in 4 of the 10 cases. There was a relationship between the degree of abnormality and the number of electrodes that could be inserted. All 10 patients can perceive sound and are active users of their implants. PMID- 7475873 TI - Otitis media following tympanostomy tube placement in children with IgG2 deficiency. AB - Children with IgG2 deficiency commonly develop recurrent acute otitis media. It is believed that these infections are secondary to impaired antibody response rather than eustachian tube dysfunction and are therefore less responsive to treatment with tympanostomy tubes. The authors compared the incidence of acute otitis media in IgG2-deficient patients following tympanostomy tube placement with controls in a retrospective cohort study. The charts of 20 patients (10 with IgG2 deficiency and 10 controls) were reviewed. Episodes of otitis media were recorded for 12 months. IgG2-deficient patients experienced three times as many occurrences of otitis media as did controls. This suggests that otitis media is much more common in these patients following tympanostomy tube placement. We believe that an immunodeficiency workup should be considered in patients with multiple episodes of otitis media following placement of tympanostomy tubes. PMID- 7475874 TI - Topoisomerase I activity in squamous cell carcinoma of the head and neck. AB - In recent preclinical and clinical trials, topoisomerase I inhibitors have shown great promise as antitumor agents. These agents are most effective against tumors with high topoisomerase I activity. Therefore, determining topoisomerase I activity in advance may predict response to topoisomerase I inhibitors. Squamous cell carcinoma of the head and neck and normal tissue samples were obtained from 12 patients. Cellular extracts were prepared, and topoisomerase I activity assays were performed. The results suggest that topoisomerase I activity in squamous cell carcinoma of the head and neck is increased approximately sixtyfold compared to normal tissue. Increased activity often correlates with clinical responsiveness; these results predict that topoisomerase I inhibitors should be effective and selective against squamous cell carcinoma of the head and neck. PMID- 7475876 TI - Mucociliary clearance abnormalities in the HIV-infected patient: a precursor to acute sinusitis. AB - An increasing number of patients with human immunodeficiency virus (HIV) suffer from acute infectious sinusitis, and many suffer recurrent episodes at a higher rate than their non-HIV counterparts. This study investigates a mechanism underlying the increased incidence of sinusitis, that of prolonged mucociliary transport time (MTT). Nasal mucociliary clearance was examined in 30 HIV-infected patients and 30 matched, non-HIV controls using a nasal saccharin transport test. MTTs for the study group and the controls were 11.9 +/- 5.9 minutes and 7.4 +/- 3.7 minutes, respectively. This difference attained statistical significance (P < .05). Study group patients with a history of sinusitis had a mean MTT of 13.7 +/- 6.8 minutes. Those with complaints of "new onset" nasal obstruction since HIV conversion had a mean MTT of 13.5 +/- 6.8 minutes. Statistical significance (P < .05) was found comparing these times to controls, as well as to study patients without these symptoms. These data support an inherent delay of mucociliary clearance in HIV-infected patients which is chronic, possibly irreversible, and, in association with nasal obstruction, represents a major mechanism of both the high acute and recurrent sinusitis rate in this population. The cause of the mucociliary delay is still unclear and needs to be further investigated. PMID- 7475875 TI - Microcystic adnexal carcinoma: management options based on long-term follow-up. AB - Microcystic adnexal carcinoma (MAC), a recently described neoplasm that frequently affects the head and neck, presents a confusing problem for the clinician due to its unusual behavior. The individual cells have a bland microscopic appearance, and there is a predilection for neural invasion. Four cases of MAC are reported. All four cases demonstrate the difficulty with pathologic diagnosis. Follow-up of as long as 33 years begins to delineate the protracted nature of MAC. In addition, this paper includes the first report of a case of lymph node metastasis. Although resection may result in a significant defect, negative margins may not be achieved. Despite this, the defect can heal, as demonstrated by the cases described. In addition, MAC may recur many years later, irrespective of the status of the margins at the time of surgery. Given these unusual characteristics and the slowly progressive nature of MAC, strong consideration must be given to less radical surgical procedures, with close follow-up for grossly recurrent disease. PMID- 7475877 TI - Neck flexion and extension in children with Down syndrome: a somatosensory study. AB - The cervical spine in Down syndrome patients is often lax, at least at radiographic imaging, and the risk of spinal cord embarrassment is increased. This study was performed to obtain information that would help identify patients at risk for this problem and reduce the risk of endotracheal intubation and other surgical procedures in patients with Down syndrome. Somatosensory evoked potential studies were performed in 15 consecutive children who were undergoing elective otolaryngic surgery. None had neurologic symptoms or the physical examination finding of cervical spinal cord embarrassment, and their cervical spines were considered normal by plain radiographs obtained in the neutral, flexed, and extended positions. No significant change in latency (P = .16) or amplitude (P = .19) was found when the anesthetized children had their necks placed in either full flexion or full extension. With more than 90% certainty, the authors believe that children with Down syndrome who have "normal" plain cervical spine radiographs are exposed to no extra risks from neck flexion or extension during surgery. PMID- 7475878 TI - Congenital tracheal cartilaginous sleeve. AB - The congenital tracheal cartilaginous sleeve (TCS) results from a vertical fusion of the tracheal cartilages. This rare malformation is usually associated with one of the craniosynostosis syndromes, such as Crouzon's disease, Pfeiffer's syndrome, or Goldenhar's syndrome. Three new cases of TCS are reported, two with autopsy findings including the histopathology of horizontal tracheal sections. Salient features of the clinical presentation, diagnostic evaluation, endoscopic findings, histopathology, treatment, and prognosis for TCS are summarized from the perspective of the otolaryngologist. In addition, the literature is reviewed, and previously reported cases are discussed. Although infants with TCS often have multiple abnormalities, this tracheal malformation is not incompatible with life. Since multiple lesions of the larynx and trachea may be present, endoscopic evaluation is recommended for infants with TCS who experience airway distress. A smooth trachea lacking the normal ridges of tracheal arches suggests the diagnosis of TCS. With early recognition and appropriate management (including tracheotomy, if necessary, and aggressive management of pulmonary hygiene), patients may survive into childhood. PMID- 7475879 TI - Gastroesophageal reflux prophylaxis decreases the incidence of pharyngocutaneous fistula after total laryngectomy. AB - Pharyngocutaneous fistula is a serious complication after laryngectomy. Gastric fluid is known to cause severe laryngopharyngeal injury and poor mucosal healing. This study was designed to evaluate the effects of a gastroesophageal reflux prophylaxis regimen on the incidence of fistula after laryngectomy. Twenty-one consecutive patients were entered in the study. Patients with positive resection margins were excluded from the overall analysis. All patients had a Connell's two layer closure of the pharynx with absorbable suture, suction drainage of the neck, postoperative tube feeding, and prophylactic antibiotics. All patients were started on an antireflux regimen postoperatively composed of continuous tube feeds, intravenous ranitidine, and intravenous metoclopramide hydrochloride. Patients were followed postoperatively with Gastrografin swallows and clinically for 8 weeks. The control group consisted of retrospectively studied patients managed identically except for the antireflux prophylaxis. The two groups were well matched for factors reported to influence the rate of pharyngocutaneous fistula formation. The control group had six fistulae (26%) and an average of 16.5 days of hospital stay. The study group had no fistulae and an average of 11.5 days of hospital stay (P = .02). This study suggests that gastroesophageal reflux may predispose to fistula formation after laryngectomy and that mechanical and pharmacological prophylaxsis decreases postoperative morbidity and length of hospital stay. PMID- 7475880 TI - Acoustic parameters and speech analysis after supracricoid hemilaryngopharyngectomy. AB - Selected characteristics of speech and voice were compared in 10 patients who had undergone supracricoid hemilaryngopharyngectomy (SCHLP) and 20 normal adult laryngeal (NAL) speakers. Durational features of tape-recorded speech samples were measured using a stopwatch. Frequency features were analyzed with the Computerized Speech Lab and the multidimensional voice program. SCHLP speech proved comparable to NAL speech in average fundamental frequency, speech rate, and group phrasing. SCHLP speech and voice were statistically less efficient than NAL speech in fundamental frequency range, jitter, shimmer, noise-to-harmonic ratio, and maximum phonation time. Age, associated cricopharyngeal myotomy, postoperative radiation therapy, and time elapsed from SCHLP completion were not statistically related to the speech and voice parameters of SCHLP speakers. PMID- 7475881 TI - Expression of a cell-cycle-associated nuclear antigen (MIB 1) in cholesteatoma and auditory meatal skin. AB - Middle ear cholesteatoma is often invasive with consequent bone destruction. Inflammatory stimulation of the underlying connective tissue, as well as an autocrine mechanism, may be responsible for the dysregulation and abnormal proliferative features of the keratinocytes in cholesteatoma. Comparative investigations were performed to assess the epithelial cell kinetics of cholesteatoma and normal auditory meatal skin. Monoclonal antibody MIB 1 immunostaining (which recognizes a nuclear antigen expressed by dividing cells) was applied using the alkaline phosphatase antialkaline phosphatase immunolabeling method. Specimens of normal auditory meatal skin (n = 7) revealed an average MIB 1 score (quotient of the MIB 1-positive cells and the total number of cells) of 7.6 +/- 2.2%. Cholesteatoma samples (n = 13) showed an average MIB 1 score of 17.4 +/- 8.9% and a heterogeneity of proliferating epithelial areas. Epithelial cones growing toward the underlying stroma exhibited high mitotic activity. Statistically, the results of this study confirm a highly significant increase in the proliferation rate of cholesteatoma keratinocytes, which had an MIB 1 score that was 2.3 times higher than the score for keratinocytes of normal external auditory meatal skin. PMID- 7475882 TI - Expressions of c-jun and p53 proteins in human middle ear cholesteatoma: relationship to keratinocyte proliferation, differentiation, and programmed cell death. AB - The c-jun protein functions as a transcription factor for many genes, and the p53 protein functions as a negative regulator of cellular proliferation, which is related to the apoptosis pathway that induces DNA damage. It has recently been shown that c-jun promotes keratinocyte proliferation and p53 induces apoptosis of various cells. In this study, the presence of c-jun and p53 in cholesteatoma was demonstrated by immunoblotting assays using polyclonal rabbit anti-c-jun antibody and monoclonal anti-p53 protein antibody, respectively. The cholesteatoma tissue incubated with anti-c-jun antibody showed the staining of keratinocytes on the basal and spinous layers of epithelium. The c-jun protein was localized in the basal layer of normal skin, and the p53 protein was present in the nucleus of keratinocytes in the granular layer of cholesteatoma epithelium. The keratinocytes of normal external ear canal skins and normal human skins were slightly stained in the granular layer of the epidermis. The present findings suggest that c-jun and p53 proteins have a role in keratinocyte differentiation, proliferation, and apoptosis in the cholesteatoma. PMID- 7475883 TI - Clinical relevance of protein-tyrosine (de-)phosphorylation in head and neck cancer. AB - Previous studies have shown that protein tyrosine (de)phosphorylation plays an important role in head and neck cancer. Protein-tyrosine kinases (PTK) and protein-tyrosine phosphatases (PTPase) activities in the cytosol of tumor tissue were significantly increased compared to normal tissue of cancer patients as well as controls. Additionally, the enzyme activities in normal tissue of tumor patients were significantly higher than enzyme activities in normal tissue of the control group. In this paper, we have correlated the cytosolic and membranous PTK and PTPase activity of tumor and nontumor tissue with several clinical and histological parameters known to influence the clinical outcome. Furthermore, we have analyzed the value of the enzyme activities as an independent predictor of clinical behavior and occurrence of second primary tumors. We confirmed our earlier observations that cytosolic and membranous PTK activities and cytosolic PTPase activities in tumor tissues are increased compared to activities in nontumor tissues and controls. Moreover, we also confirmed the findings of increased enzyme activities in nontumor tissues compared to findings in control tissues. This finding in histologically proven healthy mucosa is highly interesting because it indicates that these biochemical changes are obviously not (yet) translated into morphological changes. Significant differences were found in membranous PTK activity when the patients were grouped by sex, tumor localization, lymph node metastasis, and previous radiotherapy. During the follow up period, no relation could be found between enzyme activities in tumor and/or nontumor tissues and disease-free interval or occurrence of second primary tumors. PMID- 7475884 TI - Dynamic MRI-guided interstitial laser therapy: a new technique for minimally invasive surgery. AB - Interstitial laser therapy (ILT) is a promising therapeutic technique in which laser energy is delivered percutaneously to various depths in tissue. In this study, the authors compared high-speed magnetic resonance imaging (MRI) of ILT in tissues during treatment with post-treatment histopathologic specimens. The use of 5-second MRI scans allowed detection of thermal damage by the 1064-nm neodymium:yttrium-aluminum-garnet laser in ex vivo liver and brain tissues. These tissues were treated by ILT with 20 W of laser output for 5 to 30 seconds via a 600-microns fiberoptic inserted 1 cm into the specimens at a power density of 7 kW/cm2 at the tip of the bare fiber. Sequential MRI measurements of lesion areas made during and after treatment were compared to measurements of laser-induced tissue damage in histopathologic sections. Fast MRI scans and tissue histology both demonstrated increased lesion size with time of ILT. Serial images obtained during ILT detected thermal changes as areas of low signal intensity that exceeded the size of the post-treatment lesions as measured on either final MRI or histology. The thermal effects detectable by these high-speed MRI sequences can be used to monitor laser-induced tissue changes during therapy, thereby providing a valuable noninvasive method for the intraoperative assessment of heat distribution during ILT. PMID- 7475885 TI - Previous tonsillectomy as prognostic indicator for success of uvulopalatopharyngoplasty. AB - Factors that determine a successful outcome following uvulopalatopharyngoplasty (UPPP) for obstructive sleep apnea (OSA) are not well defined. This study was undertaken to determine if prior tonsillectomy is predictive of a lower response rate to UPPP. A retrospective review of a cohort undergoing UPPP alone or in combination with nasal septoplasty for OSA was undertaken. Preoperative and postoperative polysomnograms were obtained to evaluate the severity of the OSA. The sample was a consecutive series of 79 patients with OSA. Clinical evaluation was performed by both an otolaryngologist and a pulmonologist. Surgical treatment in this group of 79 patients included 52 UPPP and 27 UPPP in patients with prior tonsillectomy. Concurrent septoplasty was undertaken in 17 patients. Criteria for outcome were based on comparison of preoperative and postoperative polysomnograms (i.e., apnea index, respiratory disturbance index change, and lowest saturation). A response to therapy was defined as a reduction in apnea index greater than 50%. A success was defined as apnea index less than 5, reduction of respiratory disturbance index greater than 50%, and nadir saturation greater than 82%. In 79 patients with OSA, 78% responded and 37% reflected therapeutic successes. Patients with history of prior tonsillectomy were less likely to have therapeutic improvement following UPPP. In 52 patients without previous tonsillectomy, 88% responded and 52% had a successful outcome. Of those with previous tonsillectomy, 59% responded and 7% had a successful outcome. The status of previous tonsillectomy is an important prognostic indicator in the success of UPPP for the treatment of OSA. We speculate that the presence of palatine tonsils allows the removal of an extra measure of oropharyngeal tissue, thereby improving the likelihood of success for UPPP. PMID- 7475886 TI - Outpatient cautery-assisted uvulopalatoplasty. PMID- 7475888 TI - Directory of otolaryngological societies. PMID- 7475887 TI - Radiation therapy for oropharyngeal cancer. PMID- 7475889 TI - In vivo injection of antibodies directed against the cloned mu opioid receptor blocked supraspinal analgesia induced by mu-agonists in mice. AB - The intracerebroventricular (i.c.v.) injection to mice of a polyclonal antibody raised against the peptide sequence 208-216 (TKYRQGSID) of cloned rat mu opioid receptor, reduced the analgesic potency of DAMGO, morphine and beta-endorphin-(1 31) when studied 48 h later in the tail-flick test. Antinociception elicited by delta agonists, DPDPE and [D-Ala2]-Deltorphin II, and by the kappa agonist U 50488H, was fully expressed in mice undergoing this treatment. The specific binding displayed by 0.6 nM [3H]-DAMGO was reduced in membranes preincubated with the antiserum, whereas no change could be detected for 3 nM [3H]-DPDPE or 2 nM [3H]-U-69593 labelling delta and kappa opioid receptors respectively. Naloxonazine, irreversible antagonist of the pharmacologically defined mu 1 opioid receptor, and beta-funaltrexamine (beta-FNA), that also displays irreversible antagonism at mu 1/2 receptors, when injected i.c.v. 24 h before the opioids significantly reduced the activity of DAMGO and morphine. In mice treated with naloxonazine, but not with beta-FNA, the antibody further reduced the remaining analgesic effect of DAMGO and morphine. Thus, both the antibody and beta-FNA blocked a wider population of mu opioid receptors than that tagged by naloxonazine. PMID- 7475891 TI - Evaluation of age-related changes in serotonin 5-HT2 and dopamine D2 receptor availability in healthy human subjects. AB - We assessed the relation between serotonin 5-HT2 receptor availability and aging and compared it with that for dopamine D2 receptors on 19 healthy male volunteers (age range, 21-49 years) using positron emission tomography (PET) and F-18 N methylspiperone (NMS). 5-HT2 Receptor availability was obtained using the ratio of the distribution volume in the region of interest to that in the cerebellum (Bmax'/Kd' + 1). 5-HT2 Receptor measures were obtained in frontal and occipital cortices. D2 receptor availability in striatum was measured using the "ratio index". 5-HT2 Receptor availability decreased significantly with age. This effect was significantly more accentuated for 5-HT2 receptor availability in the frontal (r = 0.92, p < or = 0.0001) than in the occipital (r = 0.67, p < or = 0.0016) cortex (df = 1, p < 0.025). Dopamine D2 receptors were also found to decrease significantly with age (r = 0.63, p < or = 0.007). In a given subject, striatal D2 receptor availability significantly correlated with 5-HT2 receptor availability in the frontal (r = 0.51, p < or = 0.035) but not in the occipital cortex. These results document a decline in 5-HT2 and D2 receptor availability with age and showed an association between frontal 5-HT2 and striatal D2 receptors. PMID- 7475890 TI - Central mu opioids mediate differential control of urine flow rate and urinary sodium excretion in conscious rats. AB - Central administration of the selective mu opioid agonist, dermorphin, produces a concurrent diuretic and antinatriuretic response in conscious rats. To determine whether central mu opioids differentially affect the renal excretion of water and sodium, we examined changes in renal function produced by intracerebroventricular (i.c.v.) administration of dermorphin during continuous intravenous (i.v.) infusion of a synthetic ADH analogue in conscious Sprague-Dawley rats. During ADH infusion the typical diuresis produced by i.c.v. dermorphin was abolished although the antinatriuresis remained intact. Alone, i.v. ADH produced a decrease in urine flow rate without significantly altering urinary sodium excretion. In other studies, the effects of i.c.v. dermorphin were examined on the renal responses produced by i.v. infusion of a V2-ADH receptor antagonist. In these studies the magnitude of the V2 antagonist-induced diuresis was not altered by i.c.v. dermorphin but the increase in urinary sodium excretion produced by this antagonist was converted to an antinatriuresis. Central dermorphin did not alter heart rate or mean arterial pressure in either study. These findings suggest that the effects of central dermorphin on renal sodium and water handling are mediated by separate mechanisms; the effects on water involving changes in circulating ADH levels and the effects on sodium independent of the action of this hormone. PMID- 7475892 TI - Modulation of short term and long term memory by steroid sexual hormones. AB - Many studies have evidenced a functional interrelation between the nervous and endocrine systems in the modulation of mnemonic processes, and others have established the role played by certain hormones in these processes; however, few studies have dealt with the effects of sexual steroids on learning and memory. The aim of this work was to determine whether short-term and long-term memory is subject to hormonal modulation. Male Wistar rats, randomly assigned to 13 groups, 1 control and 12 experimental groups, were trained on a one-trial passive avoidance conditioning. The control group received saline solution, and each of the 12 experimental groups received a treatment consisted in one of following pharmacological doses of: 5, 10, 20, 30 mg of testosterone enanthate, 0.2, 0.4, 0.8, 1.2 mg estradiol valeriate, or 1, 2, 4, 6 mg norandrostenolone decanoate, respectively. All substances were applied 45 min before the training session. Retention of the learned response was tested 10 min (short-term memory) and 24 h (long-term memory) after the training session. Results indicate that under these experimental conditions the short-term memory is facilitated with testosterone enanthate at doses of 20 and 30 mg, the estradiol valeriate at doses of 0.4 mg and the nor-androstenolone decanoate at doses of 4 mg, whereas the long-term memory is facilitated with testosterone enanthate at doses of 30 mg, estradiol valeriate at doses of 1.2 mg and the nor-androstenolone decanoate at doses of 4 mg. The other studied doses were no effective. PMID- 7475894 TI - Effects of calcium channel antagonists and pertussis toxin on noradrenaline induced contractions in pulmonary artery from pulmonary hypertensive rats. AB - The influence of calcium channel antagonists, felodipine and cadmium, as well as pertussis toxin on noradrenaline-induced contractions in pulmonary artery rings from rats with pulmonary hypertension induced by monocrotaline (MCT) were examined. MCT-treated rats had pulmonary hypertension, right ventricular hypertrophy and lung oedema, as compared to corresponding vehicle-treated rats. The MCT-treated animals did not have polycythemia as compared to vehicle-treated rats. Pre-treatment of pulmonary artery rings from MCT-treated rats with felodipine and cadmium significantly reduced the maximum response without altering the EC50 or the Hill coefficient of concentration-response curve to noradrenaline. In pulmonary artery rings from vehicle-treated rats, felodipine significantly increased the EC50 and reduced the maximum response and the Hill coefficient of the concentration-response curve to noradrenaline. In contrast, cadmium did not alter these parameters in pulmonary artery rings from vehicle treated rats. Pertussis toxin did not affect noradrenaline-induced contractions in pulmonary artery rings from vehicle- or MCT-treated rats. Felodipine, cadmium and pertussis toxin were ineffective in inhibiting noradrenaline-induced contractions in aortic rings from either vehicle- or MCT-treated rats. Our results can be interpreted to indicate that alteration to voltage operated, felodipine-sensitive, calcium channels as well as, cadmium-sensitive sites contribute to the changes observed in the functional behavior of pulmonary blood vessels from pulmonary hypertensive rats. PMID- 7475893 TI - Bioenergetic and oxidative stress in neurodegenerative diseases. AB - Aging is a major risk factor for several common neurodegenerative diseases, including Parkinson's disease (PD), amyotrophic lateral sclerosis (ALS), Alzheimer's disease (AD), and Huntington's disease (HD). Recent studies have implicated mitochondrial dysfunction and oxidative stress in the aging process and also in the pathogenesis of neurodegenerative diseases. In brain and other tissues, aging is associated with progressive impairment of mitochondrial function and increased oxidative damage. In PD, several studies have demonstrated decreased complex I activity, increased oxidative damage, and altered activities of antioxidant defense systems. Some cases of familial ALS are associated with mutations in the gene for Cu, Zn superoxide dismutase (Cu, Zn SOD) and decreased Cu, Zn SOD activity, while in sporadic ALS oxidative damage may be increased. Defects in energy metabolism and increased cortical lactate levels have been detected in HD patients. Studies of AD patients have identified decreased complex IV activity, and some patients with AD and PD have mitochondrial DNA mutations. The age-related onset and progressive course of these neurodegenerative diseases may be due to a cycling process between impaired energy metabolism and oxidative stress. PMID- 7475895 TI - Orally administered opioid antagonists reverse both mu and kappa opioid agonist delay of gastrointestinal transit in the guinea pig. AB - Kappa(kappa) opioid agonists slow gastrointestinal transit in the guinea pig and the mouse but not the rat. Opioid antagonists naloxone and naltrexone are mu (mu) preferring, while the antagonist nalmefene has more kappa binding activity. When administered orally, the specific opioid antagonists naloxone, naltrexone, and nalmefene are able to reverse the gastrointestinal transit delay caused by orally administered mu and kappa opioid agonists (morphine and U-50, 488H) in a dose dependent fashion as measured by the leading edge of charcoal meal in the guinea pig. Oral naltrexone and nalmefene have significantly more central nervous system (CNS) bioavailability than oral naloxone. However, orally administered naloxone was as effective as either naltrexone or nalmefene in reversing mu opioid agonist induced orocecal transit delays (single agonist dose apparent ED50s = 12.3 +/- 4, 7.3 +/- 4, and 13.5 +/- 6 mg/kg respectively). Nalmefene was more active than either naltrexone or naloxone in its ability to reverse the kappa agonist U 50,488H (single agonist dose apparent ED50s = 18.3 +/- 12*, 37.5 +/- 5, and 61.9 +/- 5 mg/kg respectively; * = p < 0.05). These data confirm the enteric action of orally administered opioids and further supports our earlier findings of the presence of kappa opioid activity in the guinea pig enteric nervous system. PMID- 7475896 TI - Cloning of glucose transporter-3 (GLUT3) cDNA from rat brain. AB - Using PCR, glucose transporter-3 (GLUT3) was cloned in its entirety from rat brain. The similarity between human, murine and rodent glucose transporter-3 proteins exceeds 90%. Such extensive sequence conservation suggests that essentially all regions of the protein, including the transmembrane domains, are functionally important. This clone can be used to study GLUT3 expression in the rat by Northern analysis or in situ hybridization, or the sequence can be used to generate peptide antibodies. Finally, the availability of a probe for glucose transporter-3 in the rat allows studies of the regulation of GLUT3 gene expression in neurons under a variety of physiological and pathophysiological conditions. PMID- 7475897 TI - Guanidinoethane sulfate is neuroprotective towards delayed CA1 neuronal death in gerbils. AB - The potential neuroprotective effects of guanidinoethane sulfate (GES) on delayed neuronal death of hippocampal CA1 neurons were investigated using a gerbil model of forebrain ischemia. Neuronal densities of CA1 neurons in the saline control group (255.1 +/- 11.7 cells/mm) and guanidinoethane sulfate pretreated control group (249.0 +/- 9.4 cells/mm) showed no significant differences. By contrast, in animals subjected to ischemia, CA1 neurons of the guanidinoethane sulfate pretreated group showed a significantly higher number of surviving neurons (61.1 +/- 55.11 cells/mm) compared to the saline group (17.75 +/- 12.73 cells/mm) (p < 0.05, t-test). The study indicated that although partial, guanidinoethane sulfate is neuroprotective towards gerbil hippocampal CA1 neurons against ischemic insult. PMID- 7475898 TI - Minimal cardiac electrophysiological activity of alprenoxime, a site-activated ocular beta-blocker, in dogs. AB - A closed chest catheter technique was used in dogs to examine the potential cardiac effects of alprenoxime, a potent new ocular antihypertensive agent. Alprenoxime was designed to undergo metabolic activation to the beta-blocker, alprenolol, specifically within the eye using hydrolase and reductase enzymes that reside in the iris-ciliary body. Previous studies in rabbits confirmed that intraocular pressure (IOP) significantly decreased after topically instilling ophthalmic drops of alprenoxime, while heart rates remained essentially unchanged after intravenous dosing. To further explore the safety and ocular specificity of this potential antiglaucoma drug, several cardiac electrophysiologic parameters were monitored during alprenoxime infusion in anesthetized dogs. In contrast to the pharmacologically significant increases (33-144%) measured after alprenolol or other previously tested beta antagonist infusion, the identical dose of alprenoxime had no effect on sinus cycle length (SCL), conduction times through the bundle of His and atrium (H and AH), or any other monitored cardiac electrophysiologic parameter. No changes greater than 6% from baseline were detected with alprenoxime infusion. Similarly, no beta-antagonist cardiac activity could be detected in isoproterenol stimulated dogs after alprenoxime. The results demonstrate that alprenoxime has no significant cardiac activity at doses much greater than potential therapeutic levels. The study provides further support that the new agent could be safely used in treating glaucoma. PMID- 7475899 TI - Effects of testosterone on the rat renal medullary vasopressin receptor concentration and the antidiuretic response. AB - The renal concentrating ability declines with age in humans and animals. Studies suggest that the concentrating defect is due to a decrease in renal vasopressin sensitivity. With ageing, expression of the renal vasopressin V2 receptor in rat is impaired; the normal receptor expression is restored by testosterone treatment. The effect of testosterone on the renal sensitivity to vasopressin was investigated in young rats. Male rats after orchidectomy and chronic antiandrogen cyproterone acetate treatment, and female rats after chronic testosterone phenylpropionate treatment, were used. The plasma arginine-vasopressin (AVP) and testosterone concentrations, and the antidiuretic responses to AVP and the V2 agonist deamino-[8-D-arginine]-vasopressin (dDAVP) after volume loading were measured, and the renal [3H]AVP binding density was determined. The plasma AVP level decreased slightly, but not significantly, in male rats after orchidectomy and cyproterone acetate treatment, but did not alter in female rats after testosterone treatment. The AVP and dDAVP sensitivities decreased in male rats after orchidectomy and cyproterone acetate administration, and increased in female rats treated with testosterone, as compared with the animals with a normal gonadal function. [3H]AVP binding to the renal inner medullary membranes was decreased following orchidectomy or antiandrogen treatment in male rats, and increased in testosterone-treated female rats. The results suggest that testosterone may play a physiological role in maintenance of the V2 vasopressin receptor expression and hence in the normal urinary concentrating ability in rat. PMID- 7475900 TI - Scopolamine prevents the development of sensitization to methamphetamine. AB - The cholinergic neurotransmission has been implicated in various forms of neural plasticity such as kindling and learning. The present study examined the effects of scopolamine, a muscarinic cholinergic antagonist, on the development of behavioral sensitization to methamphetamine (MA). Rats treated with MA (1 mg/kg, sc) for 10 days indicated significantly enhanced motor activity when tested with MA (0.5 mg/kg) after a 7-8 day withdrawal, indicating the development of behavioral sensitization. Pretreatment with scopolamine (3 mg/kg) prior to MA administration prevented the development of the phenomenon. Rats treated with scopolamine alone showed no difference in the motor activity compared to those treated with saline. These results suggest that stimulation of muscarinic cholinergic receptors plays a role in the development of behavioral sensitization. PMID- 7475901 TI - Influence of human granulocyte-macrophage colony stimulating factor/interleukin-3 fusion protein (PIXY321) on the hematopoietic toxicity associated with anti-viral drugs (zidovudine and didanosine) in vitro using normal human marrow cells. AB - The antiviral drugs didanosine (ddI) and zidovudine (AZT), synthetic nucleoside analogs, have been used in the treatment of acquired immunodeficiency syndrome (AIDS). Although clinical use of zidovudine (AZT) is still widely used, it is associated with the development of virus disease resistance and toxicity to the hematopoietic system. Alternative nucleoside reverse transcriptase derivatives such as didanosine (ddI) have been developed in order to reduce the incidence of virus disease resistance and hematological toxicity. We report here studies designed to ev evaluate the toxicity profile comparing didanosine (ddI) with zidovudine (AZT) when used alone or in combination with normal non-adherent, T cell depleted human marrow cells plated in the presence or absence of the human cytokine fusion protein of granulocyte-macrophage colony stimulating factor and interleukin-3 (PIXY321). As expected, didanosine (ddI) was less toxic for human hematopoietic progenitor cells, i.e., CFU-GEMM, CFU-GM, CFU-Meg, and BFU-E than zidovudine. Toxicity was additive when didanosine (ddI) and zidovudine (AZT) were combined. In the absence of drugs PIXY321 colony formation was increased for all progenitor cells cultured. In the presence of didanosine (ddI) or zidovudine (AZT), either as single-agents or combined, PIXY321 reduced toxicity significantly. These results demonstrate PIXY321 is an effective cytokine capable of reversing the toxicity associated with anti-viral drugs when used in vitro where didanosine (ddI) is less toxic than zidovudine (AZT); however their suppression of hematopoietic progenitors is additive when combined. PMID- 7475902 TI - Is clozapine selective for the dopamine D4 receptor? AB - Binding of 3H-spiperone and 3H-raclopride to membranes of cells stably transfected with a human dopamine D2 receptor clone was investigated, as was that of 3H-spiperone to those stably-transfected with a human D4 receptor clone. 3H spiperone and 3H-raclopride labeled the same number of sites in the D2 receptor preparation. The inhibition of binding by clozapine, spiperone, (-) eticlopride, haloperidol and the novel substituted benzamide 1192U90 was also investigated. Clozapine and 1192U90 showed greater inhibition of 3H-raclopride binding than 3H spiperone binding to the D2 receptor. Comparison with inhibition of 3H-spiperone binding to the D4 receptor revealed that clozapine and 1192U90 displayed apparent selectivity (as assessed by Ki ratios) for the D4 receptor when compared with binding of 3H-spiperone, but not 3H-raclopride, to the D2 receptor. PMID- 7475903 TI - The synthesis and opioid receptor binding affinities of analogues of dermorphin and its N-terminal tetrapeptide fragment with dibasic acids in position 2. AB - Analysis of possible mu opioid receptor active conformations for dermorphin suggested that the topographical location of the tyramine moiety of the N terminal tyrosine can be simulated with the phenol of tyrosine or desamino tyrosine (4-hydroxyphenylpropionic acid) and a basic group located on the side chain of a dibasic acid residue located in position 2. The biological properties of respective analogs with D- or L-arginine, and D- or L-lysine in the position 2 of dermorphin or desamino-dermorphin and their N-terminal tetrapeptide fragments, has provided evidence in support of this prediction, and questions the dogma that an N-terminal tyrosine is a necessary element for opioid agonist peptides. PMID- 7475905 TI - A novel natriuretic, hypotensive factor derived from the spleen. AB - Previous experiments have shown that the reduction in plasma volume observed after administration of atrial natriuretic factor is abolished by splenectomy. In order to determine whether the spleen contains a factor that influences cardiorenal homeostasis, rat spleens were homogenized in phosphate-buffered saline, centrifuged, subjected to ultrafiltration (molecular weight cut-off 10,000), extracted on C18 affinity columns and dried. After reconstitution in isotonic saline, the extract was injected i.v. into conscious rats. In response to this extract, there was a dose-dependent diuresis, natriuresis, kaliuresis and chloruresis which plateaued during the second and third hours following administration. Solute-free water clearance was inversely related to urine output. Blood pressure fell significantly from 109 +/- 3 mm Hg to 103 +/- 3 mm Hg during the first 10 min. after injection of the extract, and tended to remain depressed thereafter. There was no accompanying increase in heart rate. We conclude that the mammalian spleen contains a natriuretic, hypotensive factor that may play a role in cardiorenal homeostasis. PMID- 7475904 TI - Vasoactive intestinal peptide increases the liberation of arachidonate from anterior pituitary cells in vitro. AB - Several secretagogues increase prolactin (PRL) release from anterior pituitary cells through biochemical pathways that involve the liberation of arachidonate from cellular phospholipids. Vasoactive intestinal peptide (VIP) increases PRL release from anterior pituitary cells through a mechanism involving the generation of cAMP. In this study, we determined whether VIP increases the liberation of arachidonate from anterior pituitary cells. Primary cultures of anterior pituitary cells were prepared from the anterior pituitary gland of female Sprague-Dawley rats. After four to five days in culture, the incubation medium was replaced with [3H] arachidonate containing medium, and the cells incubated for 90 min. The cells were then extensively rinsed with incubation medium without [3H] arachidonate to remove the [3H] fatty acid not associated with cellular phospholipids. The pituitary cells were then incubated with medium containing various concentrations of VIP and the release of [3H] arachidonate and PRL into the incubation medium determined. VIP (500 nM) significantly increased [3H] arachidonate liberation from primary cultures of anterior pituitary cells at 30 min (p < 0.5) and 60 min (p < 0.01), but had no significant effect on the liberation of this fatty acid at 15 or 120 min. PRL release was significantly increased by VIP at 30, 60, and 120 min. VIP (60 min exposure) at concentrations of 100 and 500 nM significantly increased PRL release and arachidonate liberation in a concentration-dependent manner. Similarly, VIP increased [3H] arachidonate liberation from a preparation of anterior pituitary cells enriched in lactotropes. Since the increment in [3H] arachidonate liberation was greater in the lactotrope-enriched population than in the anterior pituitary cell preparation, it is highly probable that the lactotropes are the primary source of [3H] arachidonate liberated by VIP. These experiments provide evidence that [3H] arachidonate liberation may play a role in VIP-stimulated PRL release. PMID- 7475906 TI - Effect of fasting and growth hormone (GH) administration on GH receptor (GHR) messenger ribonucleic acid (mRNA) and GH-binding protein (GHBP) mRNA levels in male rats. AB - To elucidate whether GHR and GHBP are coordinately regulated or not, we studied the effect of fasting with or without GH administration on the GHR and GHBP mRNAs in the liver as well as in extrahepatic tissues in rats. Tissues were collected from 7-week-old male rats by decapitation 1,3, and 7 days after the start of fasting. Liver GHR mRNA levels were not affected 1 day after the start of fasting but progressively decreased for the subsequent 3 and 7 days of fasting as compared with those in control rats fed ad libitum. In contrast, liver GHBP mRNA levels significantly rose after 1 day fasting, returned to the control level after 3 days and further reduced after 7 days of fasting. Changes in GHBP mRNA level after fasting were different among the tissues. A transient increase in GHBP mRNA levels was observed in muscle and heart as well as liver, while the GHBP mRNA levels in fat tissues did not change throughout 7 days of fasting. Next, bovine GH(bGH) was administered ip to the fasted rats and control fed rats for either 1 day(100 micrograms [corrected], tid) or 5 days(150 micrograms [corrected], daily). In fed rats, liver GHR mRNA level was significantly increased by 1 day bGH treatment, but after 5 days treatment with bGH it was not different from the level in saline-injected control. Accordingly, net increment of plasma IGF-I was 296.0 ng/ml with 1 day bGH treatment and 234.2 ng/ml with bGH administration for 5 days. In fasted rats, liver GHR mRNA level did not changed after 1 day treatment with bGH, but markedly decreased 5 days after bGH administration. Net increment of plasma IGF-I was slightly reduced to 284 ng/ml with 1 day treatment with bGH, and markedly decreased to 37.0 with bGH administration for 5 days. The effect of GH administration on liver GHBP mRNA level was virtually absent in either fasting or fed state. These findings suggest that GHR and GHBP mRNAs in the liver are expressed in different ways and that expression of GHBP mRNA is differently regulated among tissues. PMID- 7475907 TI - Interaction of ethanol and anoxia with muscarinic receptor--stimulated phosphoinositide metabolism during brain development. AB - The mechanism(s) by which ethanol induces alterations in brain development may involve direct actions (e.g. changes in specific biochemical pathways), or indirect effects, such as cerebral hypoxia resulting from ethanol - induced circulatory changes. Since both ethanol and hypoxia are known to affect the metabolism of phosphoinositides, which has been suggested as a possible target for ethanol's developmental neurotoxicity, in the present study we have investigated the in vitro effects of both severe hypoxia (anoxia) and ethanol (alone or in combination) on muscarinic receptor-stimulated phosphoinositide metabolism in cerebral cortex slices from neonatal rats. Anoxia markedly inhibited carbachol - stimulated phosphoinositide metabolism in adult rats (67%), but only slightly (10%) in neonatal animals. Reoxygenation reversed the effect of anoxia at both ages. On the other hand, ethanol's inhibitory effect was pronounced in neonatal rats only, and was additive to that of anoxia. The presence of ethanol did not affect the recovery of carbachol - stimulated phosphoinositide metabolism following anoxia and reoxygenation. These results indicate that ethanol and anoxia differently and independently affect muscarinic receptor - stimulated phosphoinositide metabolism and may mutually contribute to the CNS effects observed following developmental ethanol exposure. PMID- 7475908 TI - Identification of heterotrimeric GTP-binding proteins in human megakaryoblastic leukemia cell line, MEG-01, and their alteration during cellular differentiation. AB - Various heterotrimeric GTP-binding proteins (G proteins) are possible to have important functions in hematopoietic cells. However, there has been no information regarding their expression in magakaryoblasts and/or megakaryocytes. In the present study, protein contents of seven G protein alpha subunits (Gs alpha, Gi2 alpha, Gi3 alpha, Gz alpha, G11 alpha, Gq alpha and G12 alpha) and beta subunit in a human megakaryoblastic leukemia cell line, MEG-01, were analyzed by immunoblotting. Immature MEG-01 cells expressed the alpha subunits of Gs, Gi2, Gi3, Gz, G11 and G12 at protein molecule level. During the 12-O tetradecanoyl-phorbol-13-acetate (TPA)-induced differentiation process, the contents of Gi2 alpha and Gi3 alpha increased, whereas the protein levels of Gz alpha, Gs alpha, G11 alpha and G12 alpha were observed to hardly change. beta subunit was also observed to be present in immature MEG-01 cells and to increase continuously throughout the differentiation process. For the expression of Gi2 alpha and beta subunits, chronic TPA-treatment was required although Rac2, a low M(r) GTP-binding protein, was expressed abundantly by only 30 min-TPA-treatment followed by 3 day-culture. PMID- 7475909 TI - Evaluation of prolactin-like activity produced by concanavalin-A stimulated mouse splenocytes. AB - The prolactin (PRL)-like activity released into synthetic culture medium from concanavalin-A (Con-A)-stimulated mouse splenocytes was bioassayed using 3H thymidine incorporation into Nb2 lymphoma cells. At low cell density (1.0 x 10(6) cells/ml medium) Con-A-stimulated splenocytes from Balb/c mice released more PRL like activity than did splenocytes from normal C3H/HeJ mice. This difference was maintained with animals of either sex, and for animals of different ages (1, 3 and 5 months). The strain difference was observed both when splenocytes were co cultured with Nb2 cells, and when postcultured medium from splenocytes was tested. Con-A-stimulated splenocytes also released a substance into the medium which was inhibitory to PRL-induced Nb2 cell proliferation. The concentration of the inhibitory substance in the medium was related to the number of splenocytes in culture and was similar in splenocyte media preparations from both strains of mice. Polyclonal antisera to bovine PRL, rat PRL, mouse placental lactogen II, and mouse PRL did not neutralize the effect of Con-A-stimulated splenocytes to induce Nb2 cell proliferation. Bioassay of the partially purified PRL-like activity found in postcultured medium indicated that the dose-dependent induction of Nb2 cell proliferation was parallel with that of purified mPRL. Using two mouse PRL antisera (Sinha and Talamantes #118) in Western blot analyses, no antibody binding was observed to the partially-purified splenocyte PRL-like material. The data from the antisera neutralization experiments and the SDS/PAGE Western blot analyses clearly indicated that the PRL-like activity produced by Con-A-stimulated splenocytes has little homology with pituitary PRL. Further studies are required to establish the precise molecular identity of the PRL-like activity from Con-A-stimulated splenocytes. PMID- 7475910 TI - A non-radioactive fluorescent method for measuring protein kinase C activity. AB - The purpose of this study is to report the development of a non-radioactive fluorescent peptide assay for measuring protein kinase C activity (PKC). The assay is based on a glycogen synthase derived fluorescent peptide that is phosphorylated by PKC. Phosphorylation causes the peptide to migrate toward the anode while the non-phosphorylated peptide migrates toward the cathode during agarose gel electrophoresis. Quantitation of PKC activity can be accomplished by excision of the appropriate bands and measuring their relative fluorescence. Using this assay, PKC activity was measured in whole cell homogenates from cultured renal mesangial cells. The enzyme(s)-substrate system followed Michaelis Menten kinetics under limited conditions and, therefore, Lineweaver-Burk plots were used to obtain Michaelis constant and maximum velocity values. An apparent KM value of 40 microM was obtained for the fluorescent peptide substrate with a control Vmax value of 300 pmol/min. Addition of phorbol 12-myristate 13-acetate increased Vmax to 380 pmol/min. PMID- 7475912 TI - 24h withdrawal following repeated administration of caffeine attenuates brain serotonin but not tryptophan in rat brain: implications for caffeine-induced depression. AB - Caffeine injected at doses of 20, 40 and 80 mg/kg increased brain levels of tryptophan, 5-hydroxytryptamine (5-HT) and 5-hydroxyindole acetic acid (5-HIAA) in rat brain. In view of a possible role of 5-HT in caffeine-induced depression the effects of repeated administration of high doses of caffeine on brain 5-HT metabolism are investigated in rats. Caffeine was injected at doses of 80 mg/kg daily for five days. Control animals were injected with saline daily for five days. On the 6th day caffeine (80 mg/kg) injected to 5 day saline injected rats increased brain levels of tryptophan, 5-HT and 5-HIAA. Plasma total tryptophan levels were not affected and free tryptophan increased. Brain levels of 5-HT and 5-HIAA but not tryptophan decreased in 5 day caffeine injected rats injected with saline on the 6th day. Plasma total and free tryptophan were not altered in these rats. Caffeine-induced increases of brain tryptophan but not 5-HT and 5-HIAA were greater in 5 day caffeine than 5 day saline injected rats. The findings are discussed as repeated caffeine administration producing adaptive changes in the serotonergic neurons to decrease the conversion of tryptophan to 5-HT and this may precipitate depression particularly in conditions of caffeine withdrawal. PMID- 7475911 TI - Generation of antibodies against a human lipoprotein lipase fusion protein. AB - Antibodies generated against specific proteins are useful tools for studying the physiology and cell biology of the protein of interest. Although antibodies have been successfully generated against lipoprotein lipase (LPL) and used to elucidate many aspects of its biology, there have been problems with the specificity, affinity and availability of these antibodies. To circumvent these problems, we have expressed a portion of human LPL as a bacterial fusion protein. The human LPL bacterial fusion protein was utilized to generate polyclonal antibodies in rabbits that recognize intact human, rat and bovine LPL. Using these antibodies, it was possible to demonstrate a direct correlation between LPL mass and LPL activity from different samples of human post-heparin plasma. In addition, these antibodies were used to develop an ELISA for the measurement of LPL in tissue or plasma. This is a useful means for obtaining polyclonal antibodies to LPL in sufficient quantity and without contaminating mammalian proteins. PMID- 7475913 TI - Inhaled Bordetella pertussis vaccine decreases airway responsiveness in guinea pigs. AB - Bordetella pertussis (BP) has been used as adjuvant for experimental animal immunization, but its effects on airway responsiveness are uncertain. Three groups of guinea pigs were used: animals with a single exposure to inhaled BP vaccine (strain 134, total dose 1.24 x 10(12) germs), animals submitted to a sensitization procedure through inhalation of ovalbumin plus BP, and healthy control animals. Four weeks after inhalation of BP or after the beginning of sensitization, dose- or concentration-response curves to histamine were constructed in vivo and in vitro (tracheal and parenchymal preparations). We found that BP alone produced lower responses to histamine than control guinea pigs in vivo (insufflation pressure, p = 0.0003) and in tracheal tissues (p = 0.04), but not in parenchymal preparations. Sensitization did not modify the responsiveness compared with their respective controls. These results suggest that some BP component(s), probably pertussis toxin, causes a long lasting airway hyporesponsiveness in guinea pigs. PMID- 7475914 TI - In vivo effects of superantigens. AB - Superantigens are potent immunostimulatory molecules that activate both T cells and antigen presenting cells. The consequences of superantigen exposure range from induction of T cell proliferation, massive cytokine release and systemic shock to immunosuppression and tolerance. Superantigens have been directly implicated in a number of human conditions including food poisoning and toxic shock. In addition, there is evidence to suggest that superantigens are involved in the initiation of autoimmunity, and the immune dysfunction associated with HIV infection. Because of their possible role in human disease, and their potential use in immune therapy, it is important that we more completely understand the in vivo effects of superantigens. PMID- 7475915 TI - Risk of connective tissue disorders (CTDs) among women with breast implants. PMID- 7475916 TI - Effects of flavonols on P-glycoprotein activity in cultured rat hepatocytes. AB - The effects of flavonols on P-glycoprotein (Pgp) activity were studied in cultured rat hepatocytes by assessing and transmembrane transport of Rhodamine 123 (R-123) and doxorubicin (DOX). In freshly-plated hepatocytes, containing a low amount of Pgp, flavonols did not affect the cellular retention of DOX, but strongly inhibited the Pgp-mediated efflux of R-123. In 72h-cultured hepatocytes, spontaneously overexpressing functional Pgp, flavonols inhibited R-123 efflux in a dose-dependent manner, but significantly reduced DOX retention while increasing its efflux. A similar effect was found in hepatocytes obtained from rats in which Pgp was induced in vivo by 2-acetamino-fluorene (AAF) or alpha-naphthyl isothiocyanate (ANIT) treatments. These findings indicate that flavonols, dietary compounds reported to strongly upregulate the apparent activity of Pgp in cancer cell lines, may also modulate differently the transport of putative Pgp substrates in normal rat hepatocytes. The ability to affect the drug-extruding activity at the hepatocyte canalicular membrane could be of relevance to the chemopreventive action of these compounds towards liver carcinogens. PMID- 7475917 TI - Contrasting effects of peripheral decarboxylase inhibitors on plasma activity of aromatic-L-amino acid decarboxylase and semicarbazide-sensitive amine oxidase in Parkinson's disease. AB - The peripheral decarboxylase inhibitors benserazide and carbidopa, often administered in combination with L-dopa in the treatment of Parkinson's disease, are also very good inhibitors of semicarbazide-sensitive amine oxidase (SSAO). In untreated patients and in patients treated with L-dopa alone, plasma SSAO activity is normal. In patients treated with L-dopa plus benserazide or carbidopa (Madopar or Sinemnet), however, plasma SSAO activity is strongly inhibited, contrary to the paradoxical 3-fold increase in plasma aromatic-L-amino acid decarboxylase activity we reported previously. Single-dose and longitudinal studies show that the SSAO inhibition proceeds rapidly and increases even further to nearly complete inhibition after continued treatment, while aromatic-L-amino acid decarboxylase activity only transiently decreases after a single dose and increases slowly with continued treatment above pretreatment levels. Dialysis experiments confirm that the binding of benserazide to SSAO is irreversible, especially after chronic treatment. The lack of knowledge about the exact function of SSAO precludes definite conclusions about the effect of this chronic SSAO inhibition on patients. Careful follow-up studies of patients treated with Madopar or Sinemet might provide further information about the possible physiological role of SSAO. PMID- 7475918 TI - Effects of photoperiod on testicular inhibin-alpha and androgen binding protein mRNA expression during postnatal development in Siberian hamsters, Phodopus sungorus. AB - The effects of short photoperiod (SPP) on testicular inhibin-alpha and androgen binding protein (ABP) mRNA expression were investigated in Siberian hamsters during postnatal development. Hamsters were raised in either long photoperiod (LPP; 16L:8D) or in a SPP (6L:18D). Hamsters were sacrificed at the following developmental ages: preweaning (18-20 days), immature (28-30 days), pubertal (38 40 days), postpubertal (48-50 days) and adult (60-62 days; n = 7-12 animals per group). Body and testicular weights were determined and plasma FSH levels were measured by validated RIA. As expected, testicular weight and circulating FSH levels were significantly lower in hamsters raised in SPP than in LPP animals. Photoperiod had no significant effect on body weight except during postpubertal age. The northern analyses of inhibin-alpha and ABP expression in the testes revealed that the corresponding relative mRNA levels were higher in animals exposed to SPP than in animals from LPP. These findings suggest that the testes of hamsters exposed to a SPP can synthesize inhibin-alpha and ABP, and we speculate that the release of these proteins may be regulated by photoperiod and the role of FSH in the control of inhibin-alpha and ABP gene expression in male Siberian hamsters may be minimal. PMID- 7475919 TI - Augmented adrenomedullin concentrations in right ventricle and plasma of experimental pulmonary hypertension. AB - Adrenomedullin is a strong vasorelaxing peptide discovered in pheochromocytoma tissue. Monocrotaline, a pyrrolizidine alkaloid derived from Crotalaria spectabilis, is known to cause pulmonary hypertension. To test the hypothesis that adrenomedullin plays a role similar to that of atrial natriuretic peptide or brain natriuretic peptide in modulating right ventricular responses to pulmonary hypertension, we have compared concentrations of immunoreactive rat adrenomedullin and adrenomedullin messenger RNA levels in tissues of monocrotaline treated rats with those of control rats. We also measured the effect of pulmonary hypertension on plasma level of the peptide. The concentrations of adrenomedullin immunoreactivity in right ventricle and plasma were significantly higher in monocrotaline treated rats than in control rats. The messenger RNA level of adrenomedullin in right ventricle was also higher in monocrotaline treated rats than in control rats. These results suggest that adrenomedullin participates in the mechanism to counteract the high blood pressure in pulmonary circulation. PMID- 7475920 TI - Effect of stress on amino acids and related compounds in various tissues of the rat. AB - The composition of various amino acids and related compounds in the aorta, ventricle, atria, liver, kidney, pancreas, bronchi and adrenals of rats is presented. These patterns are qualitatively similar, but quantitatively different. Stress changed these patterns. In the aorta, alpha-aminobutyric acid and ammonia are decreased. In the ventricle, phosphoserine and red. Glutathione are increased; and ammonia, arginine, asparagine, carnosine, ethanolamine, glutamic acid, glutamine, lysine, phosphoethanolamine and taurine are decreased. In the atria, alpha-aminobutyric acid, aspartic acid, ethanolamine and red. glutathione are increased; and ammonia is decreased. In the liver, alpha aminobutyric acid, cystine, isoleucine, red. glutathione, methionine and phenylalanine are increased. In the kidney, ethanolamine is increased; and beta - aminobutyric acid, citrulline, cystathionine, glutamic acid, glycine and tryptophan are decreased. In the pancreas, alpha-aminoadipic acid, ox. glutathione, leucine, glutamine, 1-methylhistidine, phenylalanine, phosphoserine, tryptophan and valine are increased; and ammonia, cystine and aspartic acid are decreased. In the adrenal glands, anserine, glutamic acid, glutamine and ox. glutathione are increased; and arginine is decreased. In the bronchi, ethanolamine and beta-alanine are increased and alpha-aminobutyric acid and ox. glutathione are decreased. Thus, stress affects certain amino compounds but changes are substance and tissue specific and independent of changes seen in the plasma. PMID- 7475921 TI - Role of the kidney in the plasma clearance of angiotensinogen in the rat: plasma clearance and tissue distribution of 125I-angiotensinogen. AB - We studied the tissue distribution and plasma clearance of angiotensinogen (AGN) in rats following an i.v. injection of 125I-labeled AGN. The plasma clearance rate of [125I]AGN fits a two-compartment model with half-lives of 10.2 +/- 1.5 min and 4.1 +/- 0.5 h in non-treated rats, and the half-life of slower phase significantly increased to 10.2 +/- 1.1 h following bilateral nephrectomy. Radioactivity was predominantly distributed in the kidneys (4.9%), and to a lesser extent in the liver (1.8%), testis (1.2%), spleen (0.61%), heart (0.35%), lung (0.18%), thymus (0.03%) and brain (0.03%). The subcellular distribution of radioactivity in the kidney was 64% in the soluble fraction and 33% in the crude mitochondrial-lysosomal fraction. Sodium dodecylsulfate-polyacrylamide gel electrophoresis revealed that the radioactivity in the soluble fraction consisted of proteins corresponding to intact [125I]AGN, whereas the mitochondrial lysosomal fraction contained additional radioactive proteins with molecular weights between 18,000 and 29,000. When isolated kidney cells were incubated with [125I]AGN at 0 degree C, the radioactive binding was saturable and specific with a Kd value of 4.8 x 10(-11)M, whereas incubation at 37 degrees C resulted in the appearance of degraded products of [125I]AGN in the medium. These results suggested that circulating AGN is cleared mainly by the kidneys via receptor mediated endocytosis, which may play an important role in regulating plasma level of AGN. PMID- 7475922 TI - Antagonizing effects of VA-045 on reduced activity of rat locus coeruleus neurons following head injury or intravenous injection of clonidine. AB - Based on the finding that VA-045, a novel apovincaminic acid derivative, had improved disturbance in consciousness, we examined the effects of the drug on the electrical activity of locus coeruleus (LC) neurons in animal models of consciousness disturbance. The animal models of consciousness disturbance used in this experiment were closed head injury (CHI) and intravenous injection of clonidine. CHI as well as clonidine injection reduced the spontaneous activity of LC neurons. The reduction of the spontaneus activity of LC neurons following CHI or clonidine injection was restored by intravenous injection of VA-045. The change of LC neuronal activity induced VA-045 preceded desynchronization of EEG. These results suggest that VA-045 exerts its ameliorating effect on consciousness disturbances, at least in part, by augmenting the spontaneous activity of noradrenergic LC neurons. PMID- 7475923 TI - Strain- and sex-related differences of acetohexamide reductase activities in liver microsomes and cytosol of rats: the Wistar-Imamichi strain lacks the male specific microsomal enzyme activity. AB - A marked strain-related difference was observed among acetohexamide reductase activities in liver microsomes of male rats. The microsomal enzyme activities in the Fischer-344 (Fischer), Sprague-Dawley (SD) and Wistar strains were 2.58 +/- 0.50, 1.60 +/- 0.44 and 0.79 +/- 0.41 nmol/min/mg protein, respectively. The microsomal enzyme activities in these rat strains were much higher in males than in females, indicating that the microsomal enzyme is a male-specific enzyme. The Wistar-Imamichi (Wistar-IM) strain was found to lack the male-specific microsomal enzyme activity. In Fischer, SD and Wistar strains of testectomized male rats, the microsomal enzyme activities were significantly increased by the treatment with testosterone. However, testosterone treatment was ineffective on the microsomal enzyme activity in the Wistar-IM strain. These results suggest that Wistar-IM rats has a genetic deficiency of the microsomal enzyme. There was no strain-related difference among the cytosolic enzyme activities in male rats. The cytosolic enzyme activities in Fischer and Wistar rats were higher in females than in males. PMID- 7475924 TI - Responses to a nonpeptide angiotensin receptor agonist, L 163491, in the feline pulmonary vascular bed. AB - The mechanism by which a novel potent non-peptide angiotensin subtype 1 receptor (AT1) agonist, (5,7-dimethyl-2-ethyl-3-[[2'-[(butyloxycarbonyl) aminosulfonyl]-5' (3-methoxybenzyl)-[1,1'-biphenyl]-4-yl] methyl]-3H-imidazo [4,5-b] pyridine) (L 163,491), increased pulmonary vascular resistance was investigated in the intact chest anesthetized cat under conditions of controlled blood flow. Intralobar injections of L-163,491, in doses of 10-300 micrograms i.a., caused dose-related increases in lobar arterial pressure that were partially antagonized by an AT1 receptor antagonist, DuP 532, or by staurosporine, a protein kinase C inhibitor, in doses that antagonized pressor responses to Ang II, but not to the thromboxane A2 mimic, U46619. Responses to L 163491 were not altered by PD 123319, an AT2 receptor antagonist. These data provide support for the hypothesis that vasoconstrictor responses to L 163491 are mediated by the activation of AT1 receptors and the protein kinase C pathway in the pulmonary vascular bed of the intact-chest cat. PMID- 7475925 TI - Hirulog effect in rat endotoxin shock. AB - Hirulog is a thrombin catalytic site inhibitor which exhibits specificity for the anionic binding exosite of alpha thrombin. Here, we have evaluated the effect of Hirulog (1, 5 and 10 mg/kg, 30 min pretreatment) in a rat model of endotoxemia. Intravenous injection of lipopolysaccharide from E. coli (25 mg/kg; serotype 0127:B8) caused decreases in blood pressure which were significantly reduced (about 60%) in animals pretreated with Hirulog. Rat survival to endotoxin was significantly increased in Hirulog pretreated group (5 and 10 mg/kg) up to 24 hours. Hirulog at the dose of 10 mg/kg inhibited both endotoxin-induced leukopenia at 30 and 60 minute points and thrombocytopenia at 30 minute point but not at 90 and 120 minute points. Fibrinogen levels were significantly reduced after 2 hours following endotoxin administration. Pretreatment with Hirulog (5-10 mg/kg i.v.) 30 min prior to administration of endotoxin prevented changes in fibrinogen plasma levels. These results demonstrate that Hirulog-induced inhibition of thrombin is effective in reducing toxic and lethal effects of endotoxin. PMID- 7475927 TI - The antioxidant, U74389, ameliorates the depression of vascular reactivity caused by lipopolysaccharide. AB - There is growing evidence that an oxidant stress contributes to the deleterious effects of bacterial lipopolysaccharide (LPS). The present study evaluated the ability of the antioxidant, U74389, to prevent the depression of vascular reactivity caused by LPS. Aortic rings taken from rats given LPS showed a depression of maximum force in response to phenylephrine that was reversed by an inhibitor of nitric oxide synthase. Pretreatment of animals with U74389 attenuated this depression of vascular reactivity. U74389 did not limit the increase in serum tumor necrosis factor levels caused by LPS. These results show that U74389 can ameliorate the depression of vascular reactivity caused by LPS possibly by interfering with the induction of nitric oxide synthase. PMID- 7475926 TI - High affinity ibogaine binding to a mu opioid agonist site. AB - The naturally occurring indole alkaloid ibogaine is of interest because of its reported ability to block drug seeking behavior for extended periods. The compound also potentiates morphine-induced analgesia in mice and reduces certain naltrexone-precipitated withdrawal signs in morphine-dependent rats. Although these results might suggest ibogaine interaction with opioid receptors, previous receptor binding studies (Brain Res. 571:242-247, 1980) found that ibogaine had a Ki value of only 2 microM for the kappa opioid receptor and was virtually inactive in blocking mu and delta receptor binding (Ki > 100 microM). The present investigation of ibogaine interaction with the mu opioid receptor from mouse forebrain labeled with [3H]-naloxone, however, yielded significantly more potent mu opioid Ki values. LIGAND analysis indicated that the data were best fit by a two site binding model, with Ki values of about 130 nM and 4 microM, reflecting ibogaine recognition of different agonist affinity states of the receptor. Inclusion of 100 mM NaCl in the assay to induce the agonist low affinity state of the receptor, reduced ibogaine's inhibition of [3H]-naloxone binding. These results suggest that ibogaine is an agonist at the mu opioid receptor with a Ki value of about 130 nM, potentially explaining ibogaine's antinociceptive effects as well as its reported reduction of opioid withdrawal symptoms and attenuation of drug seeking behavior. PMID- 7475928 TI - Facilitation of norepinephrine release from cerebral cortex is mediated by beta 2 adrenergic receptors. AB - Facilitatory effects of prenalterol and albuterol (beta 1- and beta 2-selective adrenergic agonists, respectively) in the absence and presence of propranolol (a nonselective beta-adrenergic antagonist), ICI 89,406 or ICI 118,551 (beta 1- and beta 2-selective adrenergic antagonists, respectively) on electrical stimulation evoked release of 3H-NE from rat cerebral cortical slices were assessed. Albuterol (0.1-100 nM) increased evoked release of 3H-NE from the cerebral cortical slices with greater potency than prenalterol (1-100 nM). The beta 2 adrenergic antagonist ICI 118,551 (1 nM) and propranolol (50 nM) abolished the facilitatory effects of albuterol (0.1 and 10 nM). In contrast, the beta 1 adrenergic antagonist ICI 89,406 (1 nM) did not alter the release-enhancing effect of albuterol. Prenalterol (10 and 100 nM)-induced facilitation of evoked release of 3H-NE was abolished by ICI 118,551; propranolol reduced the effect of 10 nM prenalterol and abolished that of 100 nM prenalterol. ICI 89,406 inhibited the effect of 100 nM prenalterol without altering that of 10 nM prenalterol. Basal release of 3H-NE was not altered by the drugs used in this study. These results suggest that facilitation of 3H-NE release induced by beta-adrenergic agonists is mediated primarily by beta 2-adrenergic receptors. PMID- 7475929 TI - Specificity of human UDP-glucuronosyltransferases and xenobiotic glucuronidation. AB - Several human liver UDP-Glucuronosyltransferases (UGTs) have been cloned and the cDNAs expressed in heterologous cell lines. This technological advance has allowed the assessment of the functional substrate specificity of these UGTs. The problems which may be encountered with the latency and assay of UGTs are briefly described. The data accumulated to date indicate that the Km, and possibly the Vmax/Km, for individual substrates are the best parameters to assess the specificity of the enzymes towards xenobiotic molecules. The substrate specificity of seven UGTs has been summarised from the currently available information. Of these, UGT1*02 and UGT2B8 appear to be key isoforms in the glucuronidation of a wide range of xenobiotic substrates. Additional UGTs have yet to be identified and characterised and their future inclusion may provide further insights. Finally, the functional role of each UGT in vivo has to be determined. PMID- 7475931 TI - Interaction of native and asialo rat sublingual glycoproteins with lectins. AB - The binding properties of the rat sublingual glycoprotein (RSL) and its asialo product with lectins were characterized by quantitative precipitin(QPA) and precipitin inhibition(QPIA) assays. Among twenty lectins tested for QPA, native RSL reacted well only with Artocarpus integrifolia (jacalin), but weakly or not at all with the other lectins. However, its asialo product (asialo-RSL) reacted strongly with many Gal and GalNAc specific lectins-it bound best to three of the GalNAc alpha 1-->Ser/Thr (Tn) and/or Gal beta 1-->4GlcNAc (II) active lectins [jacalin, Wistaria floribunda and Ricinus communis agglutinins] and completely precipitated each of these three lectins. Asialo-RSL also reacted well with Abrus precatorius, Glycine max, Bauhinia purpurea alba, and Maclura pomifera agglutinins, and abrin-a, but not with Arachis hypogeae and Dolichos biflorus agglutinins. The interaction between asialo-RSL and lectins were inhibited by either Gal beta 1-->4GlcNAc, p-NO2-phenyl alpha-GalNAc or both. The mapping of the precipitation and inhibition profiles leads to the conclusion that the asialo rat sublingual glycoprotein provides important ligands for II (Gal beta 1- >4GlcNAc beta 1-->) and Tn (GalNAc alpha 1-->Ser/Thr) active lectins. PMID- 7475932 TI - Pressor response induced by the hippocampal administration of neostigmine is suppressed by M1 muscarinic antagonist. AB - We investigated the roles played by three muscarinic receptors (M1, M2, and M3) in the pressor response with bradycardia that followed the injection of neostigmine (5 x 10(-8) mol) into the hippocampus of anesthetized rats. These changes were blocked by the co-administration of methylatropine (5 x 10(-8) mol). The intrahippocampal injection of pirenzepine (M1 antagonist) (5 x 10(-9) - 5 x 10(-7) mol) suppressed the neostigmine-induced pressor response dose-dependently. However injection of gallamine (M2 antagonist) (5 x 10(-8) - 5 x 10(-7) mol) and of 4-DAMP (M1 and M3 antagonist) (5 x 10(-8) - 5 x 10(-7) mol) did not suppress this hypertensive response. These findings suggest that the neostigmine-induced pressor response with bradycardia is mediated through the M1 muscarinic receptor subtype. PMID- 7475930 TI - Effects of calcium channel antagonists on the induction of nitric oxide synthase in cultured cells by immunostimulants. AB - We investigated whether calcium channel antagonists would alter the induction of nitric oxide (NO) synthesis by bacterial lipopolysaccharide (LPS) alone or in combination with interferon-gamma (IFN gamma) in cultured J774 macrophages, rat vascular smooth muscle cells, rat renal mesangial cells, and rat cardiac myocytes. The induction of NO synthesis was determined by measuring nitrite, the stable end-product. The dihydropyridine calcium channel antagonists, nifedipine, manidipine, nitrendipine, benidipine, barnidipine, perdipine, and nilvadipine all reduced the LPS-induced nitrite production in a dose-dependent manner, each with a differing half-maximal inhibitory concentration, in cultured J774 macrophages. Nifedipine also inhibited nitrite production in vascular smooth muscle cells, mesangial cells, and cardiac myocytes. The half-maximal inhibitory concentrations of nifedipine were ranked as follows: smooth muscle cells < mesangial cells < cardiac myocytes. Diltiazem, at nontoxic concentrations, had no effect on the nitrite formation in the three cell types. Verapamil markedly increased the formation of nitrite in cardiac myocytes in response to LPS and IFN gamma, but not in vascular smooth muscle or mesangial cells. Exposure of cardiac myocytes to LPS and IFN gamma caused the expression of NO synthase mRNA that was significantly increased by verapamil. Thus, certain calcium channel antagonists modulate NO synthesis by altering the induction of NO synthase. PMID- 7475934 TI - Corticotropin-releasing hormone (CRH) is expressed at the implantation sites of early pregnant rat uterus. AB - We have shown previously that the epithelial cells of human endometrium produce CRH. The biological role of endometrial CRH is not yet known. Among other things, CRH appears to be involved in the inflammatory process, acting as an autocrine/paracrine proinflammatory regulator. Since the reaction of endometrium to the invading blastocyst has characteristics of an aseptic inflammatory reaction, we have hypothesized that endometrial CRH may participate in the inflammatory phenomena taking place at the implantation site of blastocyst. In the present study we demonstrate a higher content of immunoreactive (IR)-CRH and CRH mRNA in the implantation sites of early pregnant rat uterus compared to the inter-implantation regions. Specifically we have found that: a) rat uterus contained a 1.3 kb CRH transcript, similar or identical in size to that present in human placenta, b) the size of the IR-CRH present in uterine extracts was similar to authentic hypothalamic CRH, c) Northern blot analysis showed that the content of CRH mRNA in uterus at the implantation sites was 3.5 fold higher compared to that in the inter-implantation regions and finally, d) immunohistochemical localization of IR-CRH in early pregnant rat uterus revealed positive staining of the luminal epithelial cells in both implantation and inter implantation uterine regions, while decidualized stromal cells were positive only at the implantation sites. Our data suggest that endometrial CRH may play a role in the implantation of blastocyst. PMID- 7475933 TI - Effects of cocaine, cocaine metabolites and cocaine pyrolysis products on the hindbrain cardiac and respiratory centers of the rabbit. AB - Hemodynamic and respiratory effects of vertebral artery or i.v. administration of cocaine, cocaine metabolites and cocaine pyrolysis products were measured in anesthetized rabbits. Vertebral artery administration of 1 mg of cocaine produced decreases in blood pressure and heart rate and respiratory arrest. Cocaethylene (1 mg), a cocaine metabolite produced following co-administration of cocaine and ethanol, had comparable effects except that the respiratory arrest following cocaethylene had a longer duration of action than did cocaine. A decrease in blood pressure was also observed following 1 mg of norcocaine; however, unlike cocaine, norcocaine did not affect respiration. Acute tolerance was not observed to any of the effects of 1 mg of cocaine, cocaethylene or norcocaine following vertebral artery administration. None of these compounds had significant effects following i.v. administration of the same dose. The cocaine metabolites benzoylecgonine and ecgonine methyl ester were without effect by either route in doses up to 3 mg. In contrast to cocaine, the cocaine pyrolysis products anhydroecgonine methyl ester (3 mg) and noranhydroecgonine methyl ester (3 mg) produced similar effects via both routes of administration. Both compounds produced decreases in blood pressure and heart rate and an increase in respiratory rate. Anhydroecgonine ethyl ester (3 mg), a metabolite hypothetically formed from the cocaine pyrolysis product in individuals co-administering ethanol, had effects similar to the other pyrolysis products, although its effects were not as prominent via the i.v. route of administration. Acute tolerance was observed upon administration of the cocaine pyrolysis products. These results indicate that the cocaine pyrolysis products do not share a common mechanism of action with either cocaine or the cocaine metabolites. PMID- 7475935 TI - Antiulcer and gastric secretory activity of N-octanoyl gamma-aminobutyric acid. AB - A new gamma-aminobutyric acid derivative synthesised in this laboratory, N octanoyl gamma-aminobutyric acid was found to possess antinociceptive activity but no anticonvulsant activity. The effect of the compound on gastric lesions produced by aspirin, ethanol and stress in rats, was studied and the compound was found to possess effective antiulcer action. Even though, N-octanoyl gamma aminobutyric acid did not produce a significant change in the volume of gastric acid secretion, however, it decreased the acidity and peptic activity and significantly enhanced the gastric mucus secretion. PMID- 7475936 TI - Autoradiographic characterization of binding sites for [3H]NE-100 in guinea pig brain. AB - The receptor binding specificity and neuroanatomical distribution of [3H]NE-100 (N, N-dipropyl-2-[4-methoxy-3-(2- phenylethoxy) phenyl] ethylamine monohydrochloride)-labeled sigma receptor in guinea pig brain were examined using quantitative autoradiography. NE-100 potently inhibited [3H]NE-100 binding to slide-mounted sections of guinea pig brain with the IC50 value of 1.09 nM, therefore, NE-100 apparently has high affinity binding sites. Competition studies, under conditions similar to those used to visualize the receptor, yielded the following rank order of potency: NE-100 > haloperidol > DuP734 > (+)pentazocine >> (-)pentazocine. Non-sigma ligands such as phencyclidine (PCP), MK-801 and (-)sulpiride had negligible affinities for [3H]NE-100 binding sites. High densities of [3H]NE-100 binding sites displaceable by haloperidol were present in the granule layer of the cerebellum, the cingulate cortex, the CA3 region of the hippocampus, the hypothalamus and the pons. The distribution of [3H]NE-100 binding sites was consistent with that of [3H](+)pentazocine, a sigma 1 ligand. These sigma sites may possibly be related to various aspects of schizophrenia. PMID- 7475937 TI - Influence of antidepressant drugs administration on the morphine inhibitory effect in mice vasa deferentia. AB - The effect of chronic and acute antidepressant drug administration on peripheral opioid mechanisms was investigated. For this purpose, the inhibitory effect of morphine on electrically-induced contractions of mouse vas deferens was measured. Acute antidepressant administration did not induce any change in morphine inhibition. Chronic fluvoxamine and chlorimipramine treatment induced strong and significant hypersensitivity to morphine-inhibition. Chronic desipramine, desmethylclomipramine and nortriptyline treatment induced the opposite effect. These results seem to show differential opioidergic regulation depending on the monoaminergic antidepressant used. The level of this interaction remains to be elucidated. PMID- 7475939 TI - Precision-cut tissue slices: applications in pharmacology and toxicology. AB - Almost a decade has passed since the first paper describing the isolation and maintenance of precision-cut liver slices produced using a mechanical tissue slicer was published (1). Although tissue slices of various organs have been employed as an in vitro system for several decades, the lack of reproducibility within the slices and the relatively limited viability of the tissue preparations has prevented a widespread acceptance of the technique. The production of an automated slicer, capable of reproducibly producing relatively thin slices of tissue, as well as the development of a dynamic organ culture system, overcame several of these obstacles. Since that time, significant advances in the methods to produce and culture tissue slices have been made, as well as the application of the technique to several other organs, including kidney, lung and heart. This review will i) summarize the historical use of tissue slices prior to the development of the precision-cut tissue slice system; ii) briefly analyze current methods to produce precision-cut liver, kidney, lung and heart slices; and iii) discuss the applications of this powerful in vitro system to the disciplines of pharmacology and toxicology. PMID- 7475938 TI - A dopamine D3 receptor agonist, 7-OH-DPAT, causes vomiting in the dog. AB - R(+)7-hydroxy-N,N-di-n-propyl-2-aminotetralin (R(+)-7-OH-DPAT), a selective dopamine D3 receptor agonist, (0.03-0.3 mg/kg, s.c.) dose-relatedly caused emesis, whereas S (-)-7-OH-DPAT at even 1 mg/kg did not induce emesis in dogs. Apomorphine (0.03-0.3 mg/kg, s.c.) or quinpirole (0.03-0.1 mg/kg, s.c.) also caused emesis in a dose-dependent manner. The potency of R(+)-7-OH-DPAT in inducing emesis was the same as that of apomorphine and quinpirole. On the other hand, SKF-38393 (1 and 3 mg/kg, s.c.), a selective D1 receptor agonist, failed to induce emesis in dogs. The emesis induced by R(+)-7-OH-DPAT (0.3 mg/kg, s.c.) was inhibited by S(-)-eticlopride (0.01-0.1 mg/kg, s.c.), a potent D2 and D3 receptor antagonist but not by SCH-23390 (1 mg/kg, s.c.), a selective D1 receptor antagonist or clozapine (1 mg/kg, s.c.), a D4 receptor antagonist. These results indicate that dopamine D3 receptors play an important role in the genesis of emesis in dogs. PMID- 7475940 TI - Benzodiazepine binding varies with stage of estrous cycle in unwashed membranes from mouse brain. AB - The influence of the stage of the estrous cycle on binding of [3H]diazepam was examined in membranes from brains of female mice. In order to conserve endogenous factors such as progesterone, other steroids, or GABA, the assay was performed without the extensive washing procedures typically employed in measurements of benzodiazepine binding. Significant variations in the apparent maximal numbers of binding sites (Bmax) were noted during the estrous cycle in both hypothalamus and cortex. The Bmax measured in membranes from proestrus female mice was significantly higher than in membranes from mice at other stages in the estrous cycle. Variations in apparent equilibrium binding dissociation constants (Kd) were not statistically significant by stage of the estrous cycle. The demonstrated variations in binding suggest the existence of a factor which varies with the estrous cycle in female mice and modulates the activity of the GABAA receptor complex. PMID- 7475941 TI - Microdialysis as a tool to measure dietary and regional effects on the complete profile of extracellular amino acids in the hypothalamus of rats. AB - Regional and dietary-induced changes in hypothalamic extracellular amino acid concentrations were examined. Microdialysis probes were simultaneously implanted in the hypothalamic paraventricular nucleus (PVN) and the lateral hypothalamus (LH) of anesthetized rats and perfused at a rate of 2 microliters/min. Dialysates were collected every 20 min for 1 h prior to gavage of a balanced amino acid mixture (0.85 g patterned after 1 g of chicken egg albumin) and then every 20 min for 3 h after treatment. Tail vein blood samples were also collected. Marked changes in plasma levels of most amino acids were evident immediately following the amino acid gavage. In the PVN, concentrations of isoleucine, leucine, methionine and valine all increased within 40 min, whereas significant decreases in glutamine, histidine and taurine were observed in the LH. In a separate study, PVN extracellular amino acid concentrations were examined in awake, freely behaving rats following gavage of equicaloric loads of a balanced amino acid mixture, glucose (0.89 g) or water. Dialysate levels of glutamate, isoleucine, leucine, methionine, threonine, tyrosine and valine showed reliable increases after amino acid treatment, although the overall time course of these effects differed somewhat. The amino acid profile of the PVN was, in general, unaffected by glucose administration. These findings suggest that specific brain regions may respond uniquely to amino acid ingestion and further imply that dietary composition may influence the amino acid profiles of the extracellular fluid in brain. PMID- 7475943 TI - Effects of dietary alpha-linolenic acid deficiency on neuromuscular and cognitive functions in mice. AB - Mice were fed a diet deficient in alpha-linolenic acid [18:3 (n-3)] or a control diet and the effect of this deficiency was assessed by behavioral and pharmacological measurements. Three weeks before mating female mice were fed a diet containing either peanut oil poor in alpha-linolenic acid (n-3)- or peanut+rapeseed oil rich in alpha-linolenic acid (n-3)+ = controls. Pups, aged 47 to 61 days, fed the same diet as their dams, were used for behavioral experiments. Muscular function and neuromuscular coordination assessed by the traction test, the elevated rotarod test and swimming endurance were unchanged by the (n-3)- deficiency. The level of anxiety assessed by the elevated plus-maze (anxiety protocol), the light-dark transition and the neophobia tests did not differ between (n-3)- and control (n-3)+ mice. Defensive behavior was not changed by the diet. The pentobarbital-induced loss of the righting reflex had the same duration in males, females, and controls as in (n-3) deficient mice; the latency to pentobarbital-induced loss of the righting reflex was significantly shorter in females than in males but did not differ according to the diet. Mice fed the (n 3)- deficient diet showed less efficient learning in the elevated plus-maze (learning protocol) and poorer understanding of the situation (or less motivation to escape) in the low rotarod test than mice fed the control (n-3)+ diet. PMID- 7475942 TI - L-glutamate-induced changes in intracellular calcium oscillation frequency through non-classical glutamate receptor binding in cultured rat myocardial cells. AB - The effects of L-glutamate, N-methyl-D-aspartate (NMDA), kainate (KA) and L-alpha amino-3-hydroxy-5-methyl-4-isoxazole proprionate (AMPA) on intracellular Ca2+ oscillation frequency were studied in cultured rat myocardial cells. Ca2+ oscillations per minute were increased as compared to control by L-glutamate (100 microM) from 3.8 +/- 2.2 to 25.7 +/- 4.3 (p < 0.001) and the NMDA-receptor agonist, NMDA (100 microM), from 1.2 +/- 0.8 to 34.8 +/- 10.1 (p < 0.011). Increases over control frequency were also seen in response to the non-NMDA receptor agonists KA (100 microM) from 5.8 +/- 2.3 to 25.6 +/- 3.2 (p < 0.001) and AMPA (10 microM) from 3.8 +/- 1.2 to 13.3 +/- 1.8 (p < 0.001). The non competitive NMDA receptor antagonist, (+)-5-methyl-10,11-dihydro-5H dibenzo[a,d]cyclohepten-5,10-imine maleate (MK801) (10 microM), decreased the Ca2+ oscillation frequency induced by NMDA (100 microM from 36.8 +/- 12.2 to 7.2 +/- 7.2 (p < 0.05). (+/-)-2-Amino-7-phosphonoheptanoic acid (AP7), a competitive inhibitor at the NMDA receptor inhibited the increase in frequency induced by KA (100 microM) at all concentrations tested (0.1, 1.0, 10 and 100 microM). 6,7 Dinitroquinoxaline-2,3-dione (DNQX), a competitive inhibitor at non-NMDA receptors, also decreased the oscillation frequency elicited by KA (100 microM) from 35.4 +/- 9.4 to 28.2 +/- 9.8, 24.8 +/- 9.8 and 11 +/- 9.5 at concentrations of 0.1, 1.0 and 10 microM respectively. The peak amount of intracellular Ca2+ as expressed as the fluo 3 ratio, F/Frcst, was not increased by L-glutamate, NMDA or KA. These results suggest the presence of a novel glutamate receptor composed of both non-NMDA and NMDA subunits on cultured rat myocardial cells, and receptor stimulation leads to an increase in intracellular Ca2+ oscillation frequency. PMID- 7475944 TI - Nitro containing L-arginine analogs interfere with assays for nitrate and nitrite. AB - We evaluated the effect of in vivo and in vitro administration of nitro containing and nitro-deficient L-arginine-derived nitric oxide (NO) synthase inhibitors on the measurement of NO in plasma, urine and HEPES buffered physiologic salt solution (PSS) by ozone chemiluminescence and by the modified Griess reaction. In vivo administration of 1, 5, 25, 40 or 50 mg/kg of NG-nitro-L or D-arginine methyl ester (LNAME, DNAME), NG-nitro-L-arginine (LNA) or aminoguanidine (AG) to rats and mice increased NO in urine and plasma as determined by chemiluminescence using 2.3% vanadium chloride in 2N HCI at 100 degrees C as the redox reagent. In vivo administration of 1 and 10 mg/kg/day of NG-imino-ethyl-L-ornithine (LNIO) or 3 amino-1,2,4 triazine (AT) reduced plasma and urine NO. Addition of LNAME, DNAME and LNA (100 nM to 1 mM) to the redox solution produced a concentration response curve for NO in the chemiluminescence assay similar to that produced by standard solutions of sodium nitrite and nitrate. LNMMA produced a small NO signal but only at concentrations equal to or exceeding 0.1 mM. LNIO, AT and AG did not give any NO signal even at concentrations exceeding 1 mM. Conversion of plasma or urine nitrate to nitrite with cadmium gave elevated values of plasma nitrite by the Greiss assay when LNAME or LNA was the NO synthase inhibitor. We conclude that in vivo and in vitro use of LNAME and LNA and in vivo use of high doses of aminoguanidine interfere with the assay of NO2- and NO3- with the modified Griess reaction and with chemiluminescence. We suggest that LNAME and LNA not be used in vivo or in vitro when total RNI is measured with these assays. PMID- 7475945 TI - Amiodarone induced lipidosis similar to Niemann-Pick C disease. Biochemical and morphological study. AB - Amiodarone is effective in the treatment of supraventricular and ventricular cardiac arrhythmia, however a high incidence of toxic side effects has been observed in various organs and tissues during chronic treatment. Ultrastructural observation of affected tissues reveals myelinoid inclusion bodies. The exact pathogenetic mechanism of these changes is still unknown. In this study we investigated the biochemical effects of this drug on lysosomal hydrolases and the alterations induced in subcellular organelles of fibroblasts cultured for 24 h with different concentrations of amiodarone in the medium. Of the enzyme activities assayed, we only observed a significant reduction in sphingomyelinase. Ultrastructural observation of fibroblasts showed swollen lysosomes and a few onionoid inclusion bodies at lower concentrations of the drug; at higher concentrations the lysosomal system was severely impaired. Cytochemical staining of unesterified cholesterol with filipin showed accumulation of cholesterol. We conclude that chronic amiodarone treatment in experimental conditions induces inhibition in sphingomyelinase activity through interaction with membrane lipids and modification of bilayer structure. Higher concentrations of the drug impair cholesterol transport and induce lipid accumulation. These results may be useful for understanding the pathogenesis of induced lipidosis in patients in chronic treatment with amiodarone. PMID- 7475946 TI - Pentamidine does not interfere with nitrite formation in activated RAW 264.7 macrophages but inhibits constitutive brain nitric oxide synthase. AB - Pentamidine effects on the interferon-gamma- or interferon-gamma plus bacterial lipopolysaccharide-induction of nitric oxide synthase in the macrophage cell line RAW 264.7, determined by measuring nitrite release into culture supernatants, were investigated. At concentrations above 10 microM, pentamidine caused visible toxic effects including cell lysis which also was assessed by measuring lactic dehydrogenase release. A progressive inhibitory effect of pentamidine could not be clearly dissociated from these toxic and lytic effects which were extensive at 100 microM. At 1 microM pentamidine, the dose response dependence of nitrite formation on interferon-gamma was not affected. Tumor necrosis factor-alpha caused some enhancement of interferon-gamma-induced nitrite release only at high doses of 100 and 10,000 unit/ml. Pentamidine had no effect on isolated inducible nitric oxide synthase from RAW 264.7 cells but inhibited the constitutive enzyme from pork cerebellum non-competitively. The lack of any stimulatory effect of pentamidine on nitrite production in RAW 264.7 cells suggests that NOS induction and NO production by macrophages is not the mechanism of the antimicrobial effects of this drug. PMID- 7475947 TI - Reduction of acetylcholine-activated current by low concentrations of extracellular adenosine 5'-triphosphate. AB - Effects of adenosine 5'-triphosphate (ATP) on ionic currents activated by acetylcholine (ACh) were investigated using rat pheochromocytoma PC12 cells and Xenopus oocytes expressing nicotinic receptors. In PC12 cells, ATP (10 nM to 1 microM) inhibited an inward current activated by ACh in not all but about 60% of cell batches. The ACh-activated current was also inhibited by ATP in Xenopus oocytes and, with a subunit combination of alpha 3 plus beta 4, the inhibition was observed at concentration as low as 100 fM. Uridine 5'-triphosphate (UTP) induced a similar inhibition of the ACh-activated current both in PC12 cells and Xenopus oocytes. These and other properties suggest that the current reduction by ATP is not mediated through conventional P2-purinoceptors. PMID- 7475948 TI - Effects of L-name on endothelin-1-induced barrel-rolling in periaqueductal gray area of rats. AB - The injection of endothelin-1 (ET-1) into the dorsolateral periaqueductal gray (PAG) area of freely moving rats at doses from 0.1 to 1 pmol/rat induced rotation along the long axis of the body (barrel-rolling). The pretreatment of this area with L-NAME (N omega-nitro-L-arginine methyl ester, 1 mumol/rat), an L-arginine analogue and a potent inhibitor of nitric oxide (NO) biosynthesis, significantly (p < 0.01) potentiated the duration of the ET-1-induced barrel-rolling. Pretreatment of the PAG area with L-arginine (1 mumol/rat), a precursor of NO, significantly (p < 0.01) decreased the ET-1-induced effects. These preliminary data indicate that the L-arginine-NO pathway exerts a functional antagonism on ET 1 induced barrel-rolling at the level of the PAG area. PMID- 7475949 TI - Tyrosine kinase inhibitors potentiate the induction of low density lipoprotein receptor gene expression by hepatocyte growth factor. AB - The role of tyrosine kinase, protein kinase C, cyclic nucleotide- and Ca(2+) calmodulin-dependent protein kinase second messenger pathways in the induction of LDL receptor gene expression by hepatocyte growth factor (HGF) was studied in the human hepatoma cell line Hep-G2. Incubation with media containing HGF increased the level of LDL receptor mRNA by 6.5-fold. Co-incubation with HGF and either of two tyrosine kinase inhibitors genistein (2.0-20.0 micrograms/ml) and herbimycin A (0.5-500.0 ng/ml) increased the level of LDL receptor mRNA above that observed with HGF alone by 40-60%. Incubation with HGF in the presence of the calmodulin antagonist W7 (10-30 microM) also super-induced the level of LDL receptor mRNA by nearly 230%. The protein kinase C and A inhibitors chelerythrine (0.1-10.0 microM) and H8 (0.5-5.0 microM), respectively, had no significant effects on the induction of LDL receptor mRNA by HGF. Taken together, these data suggest that tyrosine kinase, protein kinases C and A, and Ca(2+)-calmodulin dependent protein kinase activities are not essential for activation of LDL receptor gene expression in Hep-G2 cells by HGF. PMID- 7475950 TI - Permeability of the blood-brain barrier to amylin. AB - Amylin is co-secreted with insulin from the pancreas of patients with non-insulin dependent diabetes mellitus, and its deposition may contribute to the central nervous system (CNS) manifestations of this disease. Amylin, but not its mRNA, is found in brain, suggesting that CNS amylin is derived from the circulation. This would require amylin to cross the blood-brain barrier (BBB). We used multiple time regression analysis to determine the unidirectional influx constant (Ki) of blood-borne, radioactively labeled amylin (I-Amy) into the brain of mice. The Ki was 8.99(10(-4)) ml/g-min and was not inhibited with doses up to 100 micrograms/kg, but it was inhibited by aluminum (Al). About 0.11 to 0.13 percent of the injected dose of I-Amy entered each gram of brain. Radioactivity recovered from brain and analyzed by HPLC showed that the majority of radioactivity taken up by the brain represented intact I-Amy. Capillary depletion confirmed that blood-borne I-Amy completely crossed the BBB to enter the parenchymal/interstitial fluid space of the cerebral cortex. Taken together, these results show that blood-borne amylin has access to brain tissue and may be involved in some of the CNS manifestations of diabetes mellitus. PMID- 7475951 TI - Distribution and translocation of isoforms of protein kinase C in rat submandibular acinar cells. AB - The distribution of six isoforms of protein kinase C (PKC) in seromucous acinar cells of rat submandibular gland was examined and their translocation from the cytosolic- to the membrane fraction after different stimuli investigated. Western blotting, immunostaining with isoform-specific antibodies and scanning densitometry showed that PKC-alpha and epsilon were distributed fairly evenly between the cytosol and membranes in resting cells, while isoforms- beta, delta and zeta were all predominantly localized (over 80%) in membranes. PKC-gamma was not detected. PKC-alpha was mobilized to the membrane fraction by the phorbol ester, TPA, but not by the phosphoinositide-coupled agonists carbachol, methoxamine and substance P (SP). PKC-epsilon was translocated by TPA and carbachol but not by SP or methoxamine. Biochemical assay of total PKC confirmed that cytosolic enzyme activity was significantly reduced by TPA and carbachol to 29% and 75% respectively of control levels. These results suggest that muscarinic regulation of the mucosecretory response in the rat submandibular gland may be mediated by the PKC-epsilon isoform. PMID- 7475952 TI - Fructus aurantii reduced portal pressure in portal hypertensive rats. AB - The purpose of this study was to investigate the effects of Fructus Aurantii (the unripe fruits of Citrus aurantium L.) on portal hypertensive rats. Portal hypertension was induced by partial portal vein ligation (PVL) in Sprague-Dawley rats. Sham-operated (Sham) rats served as controls. Hemodynamic and in vitro contractile studies were performed at 14 days after surgery. Both the aqueous extract of Fructus Aurantii and synephrine, one of its purified principles with pressor activity, were infused into the conscious PVL and Sham rats via a syringe pump. Fructus Aurantii (1.25, 2.5, & 5.0 mg/kg/min) dose-dependently reduced portal pressure in PVL and Sham rats, with the percentage change in portal pressure more pronounced in PVL rats. Mean arterial pressure was dose-dependently elevated by Fructus Aurantii. Synephrine (0.095, 0.19, & 0.38 mg/kg/min) also dose-dependently reduced portal pressure and elevated mean arterial pressure in PVL and Sham rats. Fructus Aurantii (2.8-280 micrograms/ml) induced dose dependent contractile responses mainly in aorta and mesenteric artery, but little response in portal vein. The results showed that Fructus Aurantii infusion reduced portal pressure, possibly by way of arterial vasoconstriction. PMID- 7475953 TI - Endogenous substance P inhibits the expression of corticotropin-releasing hormone during a chronic inflammatory stress. AB - We have investigated the effects of a chronic inflammatory stress on substance P (SP) levels in the hypothalami of rats given adjuvant-induced arthritis (AA). Fourteen days after injection of Mycobacterium butyricum, substance P concentrations in the paraventricular nucleus (PVN) and median eminence/arcuate nucleus were significantly increased. In AA rats injected intraperitoneally with the specific neurokinin-1 receptor antagonist RP67580, plasma ACTH and corticosterone concentrations were significantly elevated, and corticotropin releasing hormone (CRH) mRNA in the PVN was increased compared to the AA group which received saline alone. The increases in hypothalamic SP in AA, together with the data demonstrating that HPA axis activity is enhanced in AA following injection of a SP antagonist, are consistent with the hypothesis that SP is acting as an inhibitor of CRH expression in this model of chronic inflammatory stress. PMID- 7475954 TI - An in vitro bioassay for testing the direct effect of luteinizing hormone releasing hormone agonists on progesterone secretion by rat ovarian cells. AB - It is known that luteinizing hormone-releasing hormone (LHRH) and its agonistic analogs could act directly on various extrapituitary target tissues by activating their specific receptors present in these tissues. In this report, we proposed an in vitro assay method for testing the bioactivities of LHRH agonists based on their inhibitory effect on human chorionic gonadotropin (HCG)-induced progesterone secretion by rat ovarian cells. Ovaries from five to ten pseudopregnant rats (prepared by sequential hormone treatment) were mechanically and enzymatically dispersed to prepare luteal cell-rich suspensions. Addition of [D-Ala6, Pro9-Ethylamide]LHRH to ovarian cell suspensions inhibited different concentrations (1, 10 or 100 mIU/ml) of HCG-induced progesterone secretion in concentration-dependent manners. In the presence of 1 mIU/ml HCG as a stimulant, the percent inhibition of progesterone secretion by the same concentrations of [D Ala6, Pro9-Ethylamide]LHRH was most pronounced and the results were usually more readily reproducible and with less experimental variations. In this study, LHRH and twelve agonistic analogs (at 10(-9) and 10(-7) M concentrations) were compared for their inhibitory effect on 1 mIU/ml HCG-induced progesterone secretion, and their relative agonistic activities were evaluated based on their inhibition potencies. This in vitro bioassay is simple and time- and cost efficient, and can be employed as a supplementary method to the commonly utilized bioassays for LHRH agonists. This method may also be useful in screening for LHRH agonists with potent extrapituitary effects. PMID- 7475955 TI - Comparison of the effects of immobilization and pressure overload induced cardiac hypertrophy on immunoreactive beta-endorphin. AB - Acute physical stress in the form of immobilization resulted in a decrease in the concentration of immunoreactive beta-endorphin (IR-BE) in the anterior pituitary (AP) and an increase in the concentration of IR-BE in the neurointermediate lobe of the pituitary (NIL) and the plasma. Hypothalamic IR-BE was not influenced by immobilization. In response to chronic cardiovascular (physiological) stress resulting from constriction of the aorta (aortic banding) and subsequent pressure overload, the concentration of IR-BE in the AP was increased as was the concentration of IR-BE in the plasma. The concentration of IR-BE in the NIL and the hypothalamus was not affected. These findings suggest that physical stress and cardiovascular stress have the same affect on IR-BE levels in the plasma but differ in their respective effects on IR-BE in the AP and NIL and do not affect the concentration of IR-BE in the hypothalamus. The difference in the effects of each form of stress on the AP and the NIL respectively, may be attributed to either the type of stress employed (physical versus physiological), the duration of the stress (acute vs chronic), or both. PMID- 7475956 TI - Expression of constitutive endothelial nitric oxide synthase in human blood platelets. AB - Nitric oxide (NO) activates the soluble isoform of guanylate cyclase in platelets and inhibits platelet function. Several studies suggest the existence of a pathway for NO synthesis in platelets as a form of feedback inhibition, but the identity of the NO synthase (NOS) isoform present within platelets is unknown. We isolated human platelets, and synthesized cDNA from platelet RNA for analysis by PCR. Primers for human neuronal or inducible NOS failed to yield a PCR signal. However, primers specific for endothelial NOS (ecNOS) amplified a DNA band of the expected size. Analysis of nucleotide sequence revealed that the amplified DNA is ecNOS. NOS enzyme activity was detected in the platelet particulate subcellular fraction, as previously demonstrated for ecNOS in other cells. Thus, ecNOS is present in human platelets, and may play a role in the regulation of platelet function by an endogenous NO pathway. PMID- 7475957 TI - Immunoreactive inhibin decreases following bilateral ovariectomy and during the postovulatory rise of FSH in Syrian hamsters. AB - Inhibin is a heterodimeric glycoprotein which may regulate FSH synthesis and secretion as well as follicular development and maturation. The source and physiological role of inhibin have not been established for the hamster, although several investigators have suggested that this hormone may function in the regulation of FSH in this species. The major objectives of the present studies were to develop a radioimmunoassay (RIA) for the measurement of inhibin in hamster serum and tissue, to identify the primary source of inhibin and to examine the relationship between inhibin and FSH during the estrous cycle. A sensitive, accurate and specific RIA was developed and utilized to measure changes in circulating levels of immunoreactive inhibin (ir-inh-alpha) following bilateral gonadectomy and throughout the estrous cycle. Circulating ir-inh-alpha declined rapidly and significantly following bilateral gonadectomy in female hamsters suggesting a gonadal source. Serum FSH concentrations increased following the decline in serum ir-inh-alpha levels. In the adult female hamster circulating ir-inh-alpha increased gradually throughout diestrus, peaked at the time of the preovulatory gonadotropin surge, then declined to a nadir on the morning of estrus. Changes in ovarian inhibin subunit mRNAs were examined throughout the estrous cycle and correlated with changes observed in circulating ir-inh-alpha levels. Observed significant reductions in the relative amount of inhibin mRNAs and serum concentrations of ir-inh-alpha during early estrus may moderate the amount and duration of the secondary FSH rise and thus contribute to the regulation of follicle recruitment in the hamster. PMID- 7475958 TI - Isatin is a potent endogenous antagonist of guanylate cyclase-coupled atrial natriuretic peptide receptors. AB - Isatin (indole-2,3-dione) is an endogenous compound with anxiogenic properties. In the brain, highest levels (0.1 microgram/g) have been found in the rat hippocampus. In the present study, we show that isatin has little effect on a wide range of neurotransmitter and hormonal receptors but that it acts as an inhibitor of atrial natriuretic peptide (ANP) binding, with an IC50 of 4x 10(-7) M. It also inhibits ANP-activated particulate guanylate cyclase from rat kidney, heart and brain membranes in dose-dependent fashion, varying also with ANP concentration. These findings suggest that isatin is a new endogenous regulator of mammalian ANP activity, with potential implications for the control of both anxiety and natriuresis. PMID- 7475959 TI - Neuronal NK3-receptors in guinea-pig ileum and taenia caeci: in vitro characterization by their first non-peptide antagonist, SR142801. PMID- 7475960 TI - [3H]A-69024: a non-benzazepine ligand for in vitro and in vivo studies of dopamine D1 receptors. AB - [3H]A-69024 has been prepared as a radioligand for studying the dopamine D1 receptor. [3H]A-69024 binds to rat striatal membranes with a KD = 14.3 +/- 3.2 nM (mean +/- SEM; n = 3) and Bmax = 63.5 +/- 12.8 fmol/mg wet tissue (1.8 +/- 0.3 pmol/mg protein). This ligand binds to only one site with a Hill coefficient close to unity. The in vivo biodistribution of [3H]A-69024 showed a high uptake in the striatum (5.9% ID/g) at 5 min followed by clearance. As a measure of specificity, the striatum/cerebellar ratio reached a maximum of 6.7 at 30 min post-injection. Pre-treatment with the D1 antagonist R(+)SCH 23390 (1 mg/kg) reduced this ratio to unity. The dopamine antagonist (+)butaclamol and unlabeled A-69024 inhibited striatal uptake by 70 and 51%, respectively. Spiperone (D2/5 HT2A) and ketanserin (5-HT2A/5-HT2C) at doses of 1 mg/kg had no inhibitory effect on [3H]A-69024 uptake in the striatum; however, increased uptake of [3H]A-69024 by > 30% in the whole brain was observed. The selectivity and affinity of [3H]A 69024 suggests that this non-benzazepine radioligand may be useful for in vitro and in vivo studies of the dopamine D1 receptor. PMID- 7475961 TI - Alpha 2-adrenoceptors and the regulation of glucose, insulin and amylin levels in diabetic rats. AB - The effects of the selective alpha 2-adrenoceptor agonist UK 14.304 on glucose, insulin and amylin levels were examined in neonatal streptozotocin-induced diabetic rats. UK 14.304 (0.03 to 0.3 mg/kg i.p.) induced a dose-dependent reduction of both insulin and amylin levels in normal rats while UK 14.304 at 0.03 mg/kg had already a maximal effect in diabetic rats. Amylin/insulin molar ratios rose after UK 14.304 administration in diabetic rats but remainded constant in normal rats. The alpha 2-adrenoceptor antagonist deriglidole (3 mg/kg i.p.) slightly increased insulin and amylin levels in the two groups of rats but glucose levels were more markedly decreased in diabetic rats. Deriglidole completely antagonized UK 14.304 (0.1 mg/kg i.p.)-induced changes thereby normalizing amylin/insulin molar ratios in diabetic rats. These results suggest that insulin and amylin are both under inhibitory control via alpha 2 adrenoceptor though the responses may be differentially regulated. It is further suggested that diabetes in the neonatal streptozotocin-induced rat model is associated with a hypersensitivity of the pancreas to alpha 2-adrenoceptor stimulation. PMID- 7475962 TI - Roles of calcium in the regulation of apoptosis in HL-60 promyelocytic leukemia cells. AB - Increase in intracellular calcium concentrations ([Ca2+]i) is critical for the initiation of apoptosis in cells such as thymocytes and in other cells, calcium chelators may promote apoptosis. However, calcium modulators, such as calcium ionophore 4-bromo-calcium ionophore (Br-A23187) and thapsigargin (TG), induce apoptosis in different cells, including HL-60 cells in which the induction of apoptosis seems a calcium-independent process. These observations imply that the disturbance of calcium homeostasis is probably the most important factor in the regulation of apoptosis. In this article, reagents with different potencies of modulating calcium homeostasis were used to study the possible role of [Ca2+]i and the status of intracellular calcium stores in the causation of HL-60 cell apoptosis. We found that an increase in [Ca2+]i alone did not result in apoptosis, while the depletion of TG-sensitive calcium stores in the endoplasmic reticulum was closely related with the induction of apoptosis. In HL-60 cells, extracellular and intracellular calcium chelators promoted apoptosis. Calmodulin antagonist did not attenuate apoptosis induced by other reagents. Our results suggest that the depletion of Ca2+ stores is an important mean to modulate calcium homeostasis and that the mobilization of calcium (Ca2+) from intracellular stores, rather than an increase in [Ca2+]i, provides the signal for the induction of apoptosis in HL-60 cells. PMID- 7475963 TI - Neurotropin prevents neurophysiological abnormalities and ADP-induced hyperaggregability in rats with streptozotocin-induced diabetes. AB - Neurotropin, a non-proteinaceous extract from the inflamed dermis of rabbits inoculated with vaccinia virus, was administered for 8 weeks to rats with streptozotocin-induced diabetes. The physiological and biochemical changes of the nerves were studied as well as ADP-induced platelet aggregation. Neurotropin improved the caudal motor nerve conduction velocity, R-R variability, sciatic nerve blood flow, and platelet hyperaggregability in diabetic rats, despite having no effect on sorbitol and fructose accumulation or myoinositol depletion in the sciatic nerve. The correlation between nerve conduction velocity, R-R variability, nerve blood flow, and platelet aggregation were significant between each two parameters (p < 0.0001). Thus, the mechanism of action of neurotropin differed from that of aldose reductase inhibitors. These findings suggest that vascular factors may play an important role in the development of diabetic neuropathy, and that neurotropin may be useful for the treatment of this condition. PMID- 7475965 TI - The influence of testosterone on purine nucleotide metabolism in rat liver. AB - Purine nucleotide metabolism was studied in rat liver by following the incorporation of 14C-formate into soluble nucleotides, uric acid and RNA ribonucleotides. After castration, GMP formation was less than that of AMP, and purine nucleotide catabolism and RNA synthesis decreased. Testosterone administration did not modify GMP or AMP synthesis, but restored purine nucleotide catabolism and RNA production to normal values. These results demonstrate the influence of testosterone on purine nucleotide metabolism in a non-reproductive organ. PMID- 7475966 TI - Effects of thioperamide, a histamine H3-receptor antagonist, on a scopolamine induced learning deficit using an elevated plus-maze test in mice. AB - We examined the effects of thioperamide and (R)-alpha-methylhistamine, a histamine H3-receptor antagonist and an agonist, respectively, on a scopolamine induced learning deficit using an elevated plus-maze test in mice. Thioperamide alone slightly improved the learning deficit induced by scopolamine, and pretreatment with zolantidine, a histamine H2-receptor antagonist, significantly enhanced the effect of thioperamide in this test. (R)-alpha-Methylhistamine, pyrilamine, ketotifen, terfenadine, and zolantidine alone at the doses tested had no effect. Moreover, the improvement by thioperamide plus zolantidine was antagonized by pretreatment with histamine H1-receptor antagonists such as pyrilamine or ketotifen, but not by terfenadine. Thus, thioperamide improved the scopolamine-induced learning deficit through central histamine H1 receptors in mice. The present results supported the hypothesis that histamine may play an important role in learning and memory. PMID- 7475964 TI - Receptor reserve and affinity of mu opioid agonists in mouse antinociception: correlation with receptor binding. AB - In order to quantitate the extent to which opioid agonist potencies obtained in behavioral assays are determined by the apparent in vivo affinity and efficacy of the agonist, the antinociceptive effects of the mu opioid agonists morphine, fentanyl, etonitazene, and NIH 10741 were assessed before and after administration of the insurmountable mu opioid antagonist clocinnamox (CCAM) in a 55 degrees C warm-water tail withdrawal test in Swiss albino mice. Under control conditions, all four mu opioid agonists produced a full antinociceptive response with the following ED50 values (in mg/kg): morphine, 12; fentanyl, 0.47; etonitazene, 0.039; NIH 10741, 0.0051. Analysis of CCAM's effects according to Black and Leff gave the following agonist efficacy or tau values: Morphine, 4; fentanyl 15, etonitazene, 7; and NIH 10741, 59. The respective KA values were (in mg/kg): morphine, 29; fentanyl, 7.3; etonitazene, 0.22; and NIH 10741, 0.30. The major determinant of the experimentally observed ED50 values seemed to be the apparent in vivo affinity of the respective agonist and not its efficacy. KA values (expressed as mol/kg) correlated with the Ki values (in mol/l) obtained with [3H]DAMGO radioligand binding (r = 0.96 for pKA vs. pKi), although being on average 11,000-fold higher. Values for q, the available receptor fraction as determined in the behavioral experiments, correlated strongly (r = 0.96) with the q values determined by ex vivo [3H]DAMGO- and [3H]naltrexone equilibrium binding (i.e., Bmax,clocinnamox/Bmax,control), the relationship approaching unity. PMID- 7475967 TI - Endogenous natriuretic factors 3: isolation and characterization of human natriuretic factors LLU-alpha, LLU-beta 1, and LLU-gamma. AB - A low molecular weight endogenous substance believed to be responsible for extracellular fluid homeostasis in mammals has been sought for many years. Our goal is to isolate and structurally characterize this putative "natriuretic hormone." We have developed an assay using the conscious rat to measure prolonged natriuresis (Benaksas et al (1993) Life Sciences, 52, 1045-1054), the activity originally described for this putative substance. Using this assay we have identified a number of natriuretic compounds isolated from human uremic urine. The collected urine is processed by ultrafiltration (< or = 3 kDa), gel filtration chromatography (G-25) and extraction with isopropanol and diethyl ether. The organic soluble material is then subjected to sequential high performance liquid chromatography. We report here the initial characterization of two pure isolates (LLU-alpha and LLU-gamma) obtained by this method, and the structural elucidation of a third pure compound, LLU-beta 1, a natriuretic and previously unreported metabolite of the drug diltiazem. PMID- 7475968 TI - Prenatal stress prevents the desensitization of the corticosterone response to TFMPP by desmethylimipramine, but not by phenelzine, in adult male offspring. AB - Gravid female rats were subjected to one hour of restraint stress twice daily or left undisturbed from days 14-21 of gestation. Adult 105-day old male non stressed (NS) and stressed (S) offspring were treated once daily with saline, desipramine (DMI) (10 mg/kg, sc) or phenelzine (5.0 mg/kg, sc) for 14 days. Twenty-four hours after the last injection, animals were challenged with saline or 1-(m-trifluoromethylphenyl)piperazine (TFMPP) (5.0 mg/kg, sc), a serotonin1B/2C (5-HT1B/2C) agonist, and plasma prolactin and corticosterone concentrations were measured one hour later. As compared to acute saline administration, TFMPP significantly increased prolactin and corticosterone concentrations in all groups. In NS offspring, both DMI and phenelzine treatment augmented the prolactin response, but blunted the corticosterone response, to TFMPP. In S offspring, the prolactin response to TFMPP also was augmented by phenelzine or DMI treatment, whereas the corticosterone response to TFMPP was blunted during phenelzine treatment. However, DMI treatment was not able to desensitize the corticosterone response to TFMPP in the S rats. The results indicate the adaptive capacity of 5-HT systems to DMI administration was compromised in adult animals exposed to stress in utero. PMID- 7475969 TI - Effects of short term ischemia and reperfusion on coronary vascular reactivity and myocardial function. AB - Ischemia and reperfusion have been shown to cause damage to the endothelium as well as to the cardiac myocyte. Although the vasodilator response has been shown to be impaired following ischemia and reperfusion, the effect of a short period of global ischemia on the contractile response of the coronary vasculature is not clear. In the present study, coronary vasoconstriction in response to U46619, PGF2 alpha, 5-HT, and KCl was found to be depressed for at least 15 min following 15 min of in vitro global ischemia in rats hearts. Vasodilator blockers or inactivators were used in an effort to restore this depressed coronary response. Indomethacin (5 microM) was used to block production of vasodilator prostaglandins, L-NAME (30 microM) to block production of nitric oxide (NO), and adenosine deaminase (2.4 units/ml of coronary flow) to inactivate adenosine. None of these agents restored the normal coronary constrictor response following ischemia. When superoxide dismutase and catalase (both 20 micrograms/ml of coronary flow) were infused for 5 min before and after ischemia, the coronary response recovered more than 100% of its preischemic value by 15 min of reperfusion, but still remained depressed at 5 min reperfusion. These data suggest that free radicals produced during ischemia and/or reperfusion may be at least partly responsible for this temporary "stunning" of the coronary vasculature. Since the impaired contractile response was still present at 5 min reperfusion when the buffer was supplemented with oxygen radical scavengers, another mechanism must also be involved in this "stunning" process. PMID- 7475970 TI - Sensory neuropeptides modulate cigarette smoke-induced decrease in neutral endopeptidase activity in guinea pig airways. AB - Cigarette smoke (CS) inhalation stimulates C-fibers to release sensory neuropeptides which mediate airway reflex responses to prevent irritants from entering the lower airways. When CS is inhaled via the upper airways, these airway defense responses may modulate the effect of CS on airway NEP activity and related airway hyperresponsiveness. To examine this possibility, we exposed guinea pigs to 1:10 diluted mid-tar cigarette smoke 100 puffs per day for 7 days and recorded pulmonary resistance of cumulative doses of neurokinin A (NKA, 10( 12)-10(-8) mol/kg, i.v.) or methacholine (Mch, 1-50 micrograms/kg, i.v.). NEP activity in the tracheobronchi was measured using fluorometric assay. Exposure of CS alone failed to alter the dose-response to NKA or Mch compared with air control. NEP activity in the airways after CS exposure was slightly but significantly lower than that of air control. Capsaicin pretreatment 1 week before CS exposure significantly shifted the dose-response curves of NKA, but not Mch, to the left and decreased NEP activity in the airways to a greater extent compared with CS exposure alone group. Capsaicin pretreatment alone failed to alter the responsiveness to NKA or NEP activity. CS also induced a significant increase in neutrophil counts in airways. Capsaicin pretreatment enhanced the effect of CS on neutrophil recruitment. We conclude that sensory neuropeptides may have a protective role in modulation of airways NEP activity downregulation induced by CS, probably by preventing CS from entering the lower airways or the chronic release of sensory neuropeptides induced by CS providing increased amount of substrata for NEP upregulation, and therefore modify the direct effect of CS on NEP activity and related airway hyperresponsiveness. PMID- 7475971 TI - The serotonergic antidepressant nefazodone inhibits the serotonin transporter: in vivo and ex vivo studies. AB - Nefazodone HCl (Serzone) is a new antidepressant with a chemical structure unrelated to selective serotonin reuptake inhibitors (SSRIs), tricyclics, tetracyclics, or monoamine oxidase inhibitors (MAOIs). Nefazodone is active in a number of preclinical tests for antidepressant activity and shows clinical efficacy in the treatment of depression with a more favorable side-effect profile than the structurally similar antidepressant trazodone. Previous studies have shown that nefazodone is a potent antagonist of 5-HT2A receptors and binds to the serotonin transporter in vitro and in vivo. Nefazodone also binds to the norepinephrine transporter in vitro and in acute ex vivo studies. To further investigate the ability of nefazodone to modify serotonergic transmission, the ability of systemically administered nefazodone to inhibit the serotonin transporter was assessed by investigating the ability of nefazodone to prevent p chloroamphetamine- (PCA) induced depletions of cortical 5-HT concentrations. In addition, the ability of acute and subchronic nefazodone administration to inhibit ex vivo [3H]-5-HT uptake was assessed. Acute administration of nefazodone (30, 100, and 150 mg/kg) antagonized PCA-induced depletion of cortical 5-HT concentrations in a dose-dependent manner at 1, 2, and 3 hours post-treatment. This effect was directly correlated with serum nefazodone concentrations. Both 100 mg/kg and 150 mg/kg of nefazodone were equipotent with fluoxetine (10 mg/kg) over the course of the experiment with respect to sparing of 5-HT depletion. Acute administration of nefazodone (100 and 150 mg/kg s.c.) significantly increased the Km for [3H]-5-HT uptake in rat cortical synaptosomes from 60 nmol/L in controls to 230 and 242 nmol/L in nefazodone-treated rats, respectively. Subchronic administration of nefazodone (100 and 150 mg/kg, s.c., b.i.d. x 5.5 days) reduced [3H]-5-HT uptake by 24% and 29%, respectively. Sub-chronic dosing with fluoxetine (5 mg/kg, s.c., b.i.d. x 5.5 days) reduced [3H]-5-HT uptake by 65%. These experiments confirm and extend previous reports concerning the ability of nefazodone to inhibit the 5-HT transporter in vivo. PMID- 7475972 TI - Presence of serotonin1A (5-HT1A) receptor mRNA without binding of [3H]-8-OH-DPAT in peripheral blood mononuclear cells. AB - The authors investigated the presence of serotonin receptor type 1A (5-HT1A) as labeled by the specific ligand 3H-8 hydroxy-2-(di-N-propylamino)tetralin (3H-8-OH DPAT) in saturation experiments, and the expression of the mRNA encoding them, in human peripheral blood mononuclear cells (PBMC). In situ hybridization experiments were performed as well. The results, showing that the binding of [3H] 8-OH-DPAT to lymphocyte membranes increased linearly up to 100 nM without reaching saturation, may indicate that the 3H-8-OH-DPAT was not specifically labeling the 5-HT1A receptor. By contrast, the expression studies revealed 5-HT1A mRNA in PBMC. These findings suggest that, despite the presence of mRNA, 5-HT1A receptors are not expressed in PBMC, at least in healthy controls. PMID- 7475974 TI - Interaction of abecarnil, bretazenil, and RO 19-8022 with diazepam-sensitive and insensitive benzodiazepine sites in the rat cerebellum and cerebral cortex. AB - Abecarnil, bretazenil, and Ro 19-8022 inhibited the binding of [3H]Ro 15-4513 to diazepamsensitive and -insensitive sites in the rat cerebellum and cerebral cortex, but all three had a much higher affinity for the diazepam-sensitive sites in both tissues. The GABA-shift for bretazenil and Ro 19-8022 was low ( < 2) for all sites studied, consistent with their partial agonistic profile. The GABA shift for abecarnil was appreciably higher for diazepam-sensitive binding in the cerebellum than in the cerebral cortex (1.97 vs 1.18). Furthermore, the GABA shift for abecarnil was markedly different for the diazepam-sensitive and insensitive sites in the cerebellum (1.97 vs 0.71). All three compounds inhibited [3H]Ro 15-4513 binding to the diazepam-sensitive sites with a slope factor > 1, suggestive of positive cooperativity in their binding to GABAA receptors. Abecarnil only partially inhibited diazepam-insensitive binding of [3H]Ro 15-4513 in the cerebellum, indicating that this site can be differentiated into abecarnil sensitive and -insensitive components. PMID- 7475973 TI - Chronic exercise enhances endothelium-mediated dilation in spontaneously hypertensive rats. AB - This study investigated the effect of physical training on endothelial function in spontaneously hypertensive rats (SHR). After 3 months, in conscious trained- and untrained-SHR body weight, systolic blood pressure and heart rate were 220 +/ 6 g vs 271 +/- 9 g, 172 +/- 7 mmHg vs 210 +/- 8 mmHg and 314 +/- 10 vs. 348 +/- 12 beats/min, respectively. In vitro, the dose-response curves of norepinephrine in isolated intact aortic and mesenteric rings form the exercise trained-SHR were significantly lower than those from the untrained-SHR. With denuded preparations, norepinephrine concentration-response curves were shifted to the left both in the trained- and untrained-SHR. This shift in the trained-SHR exceeded that in the untrained-SHR. The vasodilator response to acetylcholine in the trained-SHR was significantly greater than that in the untrained-SHR. Either N omega-nitro-L arginine (100 mumol/l) or methylene blue (10 mumol/l) inhibited acetylcholine induced vasodilator effect in aorta of trained- and untrained-SHR, but not in mesenteric artery of trained-SHR. Tetraethylammonium (10 mmol/l) inhibited significantly the N omega-nitro-L-arginine and methylene blue-resistant relaxation in mesenteric artery of trained-SHR, but not only by indomethacin (10 mumol/l). Collectively, these data demonstrate that chronic exercise increases EDRF/EDHF production (presumably by increasing endothelial shear stress), and may contribute to the enhanced effects of post-exercise hypotension. PMID- 7475975 TI - Astrocyte membrane Na+, K(+)-ATPase and Mg(2+)-ATPase as targets of organic solvent impact. AB - Astrocytes serve to maintain the proper homeostatic environment for neurons. In these regulations, the astrocyte membrane ATPase plays an important role. In the present study, the astrocyte were cultured in the modified Minimum Essential Medium (MEM), and the isolated cell membranes were exposed to solvents at different concentrations (3, 6 and 9 mM) for one hour. The activities of the membrane-bound Na+, K(+)-ATPase and Mg(2+)-ATPase were determined after exposure to aromatic, alicyclic and aliphatic hydrocarbons. Aromatic hydrocarbons decreased the enzyme activities dose-dependently according to their lipid solubilities. The slight enzyme-inhibiting effect of aliphatic and alicyclic hydrocarbons was not dependent on the lipophilicity of these solvents. Their molecular structure may have determined the solvent impact. In the present study, the activity of ouabain sensitive Na+, K(+)-ATPase in astrocytes was only 20-15% of the total ATPase activity, and in neurons 45-55%. The enzyme inhibition caused by organic solvents may disturb the homeostatic regulatory functions of astrocytes and thus have a toxic effect on the CNS. PMID- 7475976 TI - Cerebrospinal fluid corticotropin-releasing hormone in neurodegenerative diseases: reduction in spinocerebellar degeneration. AB - Levels of corticotropin-releasing hormone (CRH) in cerebrospinal fluid (CSF) were examined in patients with spinocerebellar degeneration (SCD) including olivopontocerebellar atrophy (OPCA), dentatorubropallidoluysian atrophy (DRPLA) and Friedreich's ataxia, Parkinson's disease (PD) and senile dementia of the Alzheimer type (SDAT), and normal aged subjects. CRH concentrations in CSF were significantly reduced in SCD compared to SDAT, PD and CSF and normal aged subjects. It is likely that degeneration not only of the cerebral cortex and the limbic system but also of the subcortical structures such as the brainstem and the cerebellum alters levels of CRH in CSF. Together with the recent anatomical and physiological evidence, the results suggest pathophysiological relevance of CRH for the cerebellar symptoms in SCD. PMID- 7475977 TI - Sodium pivalate reduces cardiac carnitine content and increases glucose oxidation without affecting cardiac functional capacity. AB - This study determined how selected functional, metabolic, and contractile properties were impacted by sodium pivalate, a compound which creates a secondary carnitine deficiency. Young male rats received either sodium pivalate (20 mM, PIV) or sodium bicarbonate (20 mM, CONTR) in their drinking water. After 11-12 weeks cardiac function and glucose oxidation rates were measured in isolated, perfused working heart preparations. Hearts were also analyzed for carnitine content, activities of hexokinase (HK), citrate synthase (CS), and B-hydroxyacyl CoA dehydrogenase (HOAD), and myosin isoenzyme distribution. Sodium pivalate treatment significantly reduced cardiac carnitine content and increased glucose oxidation but did not alter cardiac functional capacity. HK activity was increased in the PIV group (p < 0.05), and HOAD activity decreased (p < 0.05). CS activity and myosin isoform distribution (VI > 85%) remained unchanged. These results demonstrate that pivalate treatment of this duration and the accompanying carnitine deficiency shift cardiac substrate utilization without compromising cardiac functional capacity. PMID- 7475978 TI - Alpha-methyl-proline restores normal levels of bone collagen Type I synthesis in ovariectomized rats. AB - Collagen type I is the major protein of bone matrix and significantly reduced in osteoporosis. We tested the effect of alpha - methyl - proline on collagen synthesis in the model of the ovariectomized rat. Collagen synthesis was studied at the transcriptional level using Northern and dot blotting and at the protein level using hydroxyproline determination and a specific dye binding collagen assay. Alpha - methyl- proline treatment significantly increased collagen synthesis as compared to untreated ovariectomized and estradiol treated ovariectomized rats and restored collagen synthesis to levels of sham operated rats. Proline analogues were described to stimulate procollagen synthesis at the transcriptional level, however, if incorporated, lead to negative collagen production due to rapid intracellular degradation of the deficient collagen. Our synthesized analogue is not being incorporated, thus not interfering with collagen conformation and can therefore induce collagen production. PMID- 7475979 TI - Characterization of the human oxytocin receptor stably expressed in 293 human embryonic kidney cells. AB - The human oxytocin (OT) receptor was stably expressed in 293 embryonic kidney cells (293/OTR), characterized pharmacologically and compared to human uterine myometrial receptors. The cloned receptor is expressed at a reasonably high density (0.82 fmole/microgram protein) and exhibits high affinity for [3H]OT (Kd = 0.32nM), similar to the value found in human myometrial tissue. The rank-order of potency for various antagonist and agonist ligands from several structural classes is also similar between the cloned and native receptor, as seen in a comparison of their inhibitory constants for [3H]OT binding. Agonist affinity at the cloned OT receptor is decreased by guanine nucleotide analogs, demonstrating functional G-protein-coupling. The OT receptor in 293 cells, like in human myometrium, is also coupled to the inositol phosphate pathway. In 293/OTR cells, OT stimulates inositol phosphate accumulation with an EC50 = 4.1 nM, an effect blocked by a potent and selective OT antagonist, L-366,948. Additionally, the cloned receptor in 293 cells desensitizes to high concentrations of OT, similar to the desensitization in myometrial tissue and also described for several other G-protein-coupled receptors. These results illustrate the utility of the 293 cell line for expressing human OT receptors in an environment quite comparable to the native myometrial tissue. PMID- 7475980 TI - The rat model of tardive dyskinesia: relationship between vacuous chewing movements and gross motor activity during acute and long-term haloperidol treatment. AB - Tardive dyskinesia (TD) is a serious side-effect of neuroleptic treatment. In order to describe and analyse more thoroughly the rat model of TD, the behavior of the rats during cage testing was studied after acute and during long-term haloperidol (HAL) treatment. Rats were injected with HAL i.p. in an acute experiment, and in a long-term experiment, rats were treated for 4-12 months with HAL decanoate IM. Control rats received saline or sesame oil. The behavior was videotaped one h after the i.p. injection in the acute experiment, and at intervals during the long-term experiment. The putative TD analogue vacuous chewing movements (VCM), the general behavior and the type of behavior occurring simultaneously with VCM, were scored. Long-term ( > 4 months) HAL treatment increased VCM but did not change the general behavior. The single i.p. injection of HAL markedly reduced locomotion in addition to increasing VCM. Both in the acute and in the long-term experiment, VCM appeared more frequently when the gross motor activity was low, indicating an intrinsic incompatibility between gross motor activity and VCM. However, in the long-term experiment, the distribution of VCM in the different categories of behavior was the same in OIL and HAL treated rats. This shows that cage-observed VCM in rats induced by long term HAL treatment cannot be an artifact due to reduced locomotion. Thereby, an important argument against cage-observed VCM as a rat model of TD seems to be disproved. PMID- 7475982 TI - Glimepiride-induced prevention of diabetes and autoimmune events in the BB rat: revised. AB - The purpose of this study was to determine whether Glimepiride, an oral sulfonylurea drug, prevents the onset of diabetes in diabetic prone BB rats. S750181, a sulfonylurea drug that has minimal in vivo glucose metabolic effects, was also tested. In addition, the shortest period of sulfonylurea treatment required for prevention was determined. Eighty rats were studied for all treatment periods with 40 receiving a daily oral gavage dosage of glimepiride and 40 receiving a daily oral gavage dosage of vehicle solution. Diabetes onset was monitored by glycosuria and blood glucose levels. In study I, with a treatment period of 35-142 days of age, Glimepiride-treated rats showed a 32% incidence of diabetes, whereas control rats had a diabetes incidence of 55% (p < 0.04). In study II, with a treatment period of 60-140 days of age, Glimepiride-treated rats showed a 29% incidence of diabetes compared to 54% in controls (p < 0.03). Further, comparing the time of diabetes onset between the Glimepiride and control groups showed that Glimepiride delays diabetes onset (p < 0.02). In study III, with a treatment period of 60-100 days of age, Glimepiride-treated rats showed a 17% overall diabetes incidence at 170 days, whereas the controls were 43% (p < 0.01). In study IV, with a treatment period of 60-140 days of age, S750181 treated rats showed a 38% diabetes incidence and the control group showed a 43% diabetes incidence. There was no significant delaying or prevention effect observed in the S750181 group. To determine if Glimepiride affected autoimmune events, the severity of islet inflammation was examined. In study I, islet histology from total and nondiabetic animals indicated that Glimepiride-treated rats had a lower severity of islet inflammation than that of the control rats (p = 0.023). These studies show that a) Glimepiride has diabetes preventive effects, b) shorter treatment periods of only 40 days can be effective and c) Glimepiride decreases the severity of islet inflammation. PMID- 7475981 TI - Tumor necrosis factor-alpha and nitric oxide production in endotoxin-primed rats administered carbon tetrachloride. AB - Tumor necrosis factor-alpha (TNF alpha) is elevated in the sera of rats administered non-lethal doses of carbon tetrachloride (CCl4) followed by endotoxin. Elevated TNF alpha levels are correlated with the increased release of hepatic enzymes indicating hepatic damage. Under these conditions, nitric oxide (NO) was also produced in the liver as evidenced by the formation of nitrosyl complexes which were measured by electron paramagnetic resonance (EPR) spectroscopy. Decreased nitrosyl complex formation occurred in livers following treatment with either an inhibitor or macrophage activation (gadolinium trichloride; GdCl3), an inhibitor of cytokine responses (dexamethasone) or a NO synthase inhibitor (NG-monomethyl-L-arginine; 1-NMA), GdCl3 or dexamethasone treatment decreased, while 1-NMA treatment increased, TNF alpha serum level. Taken together, these data suggest that TNF alpha and NO are induced following CCl4 and LPS exposure and may be important regulators in the hepatotoxicity of this liver injury model. PMID- 7475983 TI - Gas chromatography/tandem mass spectrometry as an analytical tool for the identification of fatty acids. AB - Structures of fatty acids present at very low quantities in mycobacteria are difficult to determine. A commonly used strategy is to introduce heteroatoms into functional groups by chemical means before subjecting them to gas chromatography/tandem mass spectrometry (GC/MS/MS) analysis. Routinely used methods give very low abundance diagnostic ions leading to ambiguities in structural conclusions. GC/MS/MS associated with electron capture ionization of pentafluorobenzyl esters was used to study very complex mixtures of fatty acids from Mycobacterium fallax and M. aurum. The charge-remote fragmentation of fatty acid carboxylate anions was used for structure determination at the nanogram level of a large number of unsaturated, branched, and cyclopropane-containing fatty acids. Some of them have not been observed previously in these Mycobacteria. On the basis of these studies, biosynthetic pathways of unsaturated, branched, and cyclopropane-containing fatty acid are proposed. PMID- 7475984 TI - Peroxisomes in mice fed a diet supplemented with low doses of fish oil. AB - The influence of low dietary doses (0.1 and 0.8% w/w) of a commercial fish oil preparation on peroxisomes in normal mice was studied and compared to the known strong inductive effects of high (10%) fish oil diets. Low fish oil doses were chosen to supply the mice with a concentration of docosahexaenoic acid, which was beneficial to patients with a peroxisomal disease. Peroxisomes were evaluated by cytochemical, morphometric, and enzymological techniques. The 0.1% fish oil diet had no effect on peroxisomes in liver, heart, and kidney even after prolonged treatment. The 0.8% diet did not change the peroxisomal number nor the catalase (EC 1.11.1.6) activity in the liver. Hepatic peroxisomal beta-oxidation, however, was increased by 50% after 14 d. This was accompanied by reduced peroxisomal size. The 0.8% diet also caused a small increase (+25%) in myocardial catalase activity. No effect was observed in kidneys. Our results indicate that in mice a low (< 0.8%) dietary fish oil dose has no or only a slight effect on hepatic peroxisomal beta-oxidation. This may be of particular interest to patients with a peroxisomal fatty acid beta-oxidation defect and who display a severe deficiency of docosahexaenoic acid--diets supplemented with low fish oil doses will improve the docosahexaenoic acid level without adding a strong load to the disturbed fatty acid metabolism. PMID- 7475985 TI - Enteral feeding a structured lipid emulsion containing fish oil prevents the fatty liver of sepsis. AB - Fish oils (FO) have been shown to reduce plasma triglycerides (TG). In this study we evaluated whether enteral feeding with a structured lipid emulsion (SLE) containing FO and medium-chain triglycerides (MCT) would prevent the hypertriglyceridemia and fatty infiltration of the liver that develops during sepsis. For five days, male Lewis rats (275-300 g) were fed intragastrically a nutritionally complete diet containing a SLE or a similar diet with a soybean oil emulsion (SOE) in place of the SLE. On the fifth day, sepsis was induced by intravenously injecting 8 x 10(7) live Escherichia coli colonies/100 g b.w.; 24 h later the control SLE, septic SLE, control SOE, and septic SOE rats were sacrificed. Diet, but not treatment, had a significant effect on serum TG and free fatty acids (FFA). Feeding the SLE reduced the plasma FFA of the control and septic rats by more than 50% in comparison to both control and septic rats fed the SOE. Soleus muscle activity of lipoprotein lipase from the septic SLE rats was 44% higher than the control SLE rats. Soleus muscle from the septic SLE rats also had a twofold greater activity of lipoprotein lipase than the septic SOE rats. TG did not accumulate in the livers of the septic rats fed SLE when compared to the control SLE rats and the rats fed the SOE. Livers from the septic rats fed the SLE had a third of the TG that were present in the livers from the septic rats fed the SOE.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7475987 TI - Phospholipid molecular species composition of developing fetal guinea pig brain. AB - Adequate accumulation of polyunsaturated essential fatty acids, in particular docosahexaenoic acid (22:6n-3), into membrane phospholipids is critical for optimal fetal brain development. This process is maximal during the period of rapid neurite outgrowth, neuritogenesis, which precedes the major growth phase, myelination. There is no information about differential changes during gestation to individual brain phospholipid molecular species which contain 22:6n-3. Such details of brain development would be concealed by total fatty acid analysis of isolated phospholipid classes. We have detailed phosphatidylcholine (PC) and phosphatidylethanolamine (PE) molecular species compositions in developing fetal guinea pig brain. Total brain PC concentration increased substantially between 40 and 68 (term) d of gestation, corresponding to myelination, while PE increased in a biphasic manner between 25-35 d, which was coincident with onset of neuritogenesis, and 40-68 d. Fetal brain development was accompanied by complex changes in the concentration of individual phospholipid molecular species. During early gestation (25-40 d) 22:6n-3 was enriched in both PC and PE sn-1 16:0 molecular species. However, between 40 d and term there was no further increase in brain PC 22:6n-3 content, while brain PE was significantly enriched in both PE18:1/22:6 and PE18:0/22:6. We hypothesize that accumulation of 22:6n-3 into sn 1 18:1 and 18:0 species represents establishment of a 22:6n-3-containing membrane PE pool which may be turned over more slowly than sn-1 16:0 species. Identification of specific changes in membrane phospholipids which are associated with defined events in brain development may provide a basis for assigning functional roles to individual molecular species. PMID- 7475986 TI - Effect of tumor necrosis factor-alpha on triglyceride and phospholipid content and fatty acid composition of liver and carcass in rats. AB - We have previously shown that a continuous infusion of tumor necrosis factor alpha (TNF-alpha) in rats results in an increase in plasma triglyceride (TG), liver protein and DNA, and at the same time a reduction in muscle protein. However, there is no information on the associated effects of TNF-alpha on liver and muscle lipids. The present study, therefore, determined the effect of TNF alpha on the TG and phospholipid (PL) content and their fatty acid composition, in the liver and carcass of rats and correlated with the plasma levels of insulin, corticosterone, and catecholamines. Total parenteral nutrition that met the daily nutrient requirements was continuously infused for 10 d through a central vein catheter in two groups of rats, one receiving infused TNF-alpha (100 micrograms/kg/d) and a control group. Hepatic TG and PL, expressed either as mg/g dry weight or total organ content, were significantly increased in the TNF-alpha infused group compared with controls. Livers of TNF-alpha infused rats contained significantly less saturated and monounsaturated fatty acids and significantly more polyunsaturated fatty acids (PUFA) of the omega 3 and omega 6 series than controls. The carcass, on the other hand, contained significantly more monounsaturated and significantly less polyunsaturated fatty acids of the omega 6 series than controls. There were no changes in corticosterone level. Although plasma glucose levels were comparable between control and TNF-alpha infused rats, the latter had a marked increase in insulin levels, demonstrating insulin resistance. In addition TNF-alpha infused rats had raised norepinephrine levels.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7475988 TI - Very long chain polyunsaturated fatty acids in the blubber of ringed seals (Phoca hispida sp.) from Lake Saimaa, Lake Ladoga, the Baltic Sea, and Spitsbergen. AB - Blubbers of four ringed seal subspecies from Lake Saimaa, Lake Ladoga, the Baltic Sea, and Spitsbergen were analyzed for very long chain polyunsaturated fatty acids (VLCPUFA; > C22) using gas-liquid chromatography and gas chromatography/mass spectrometry. The VLCPUFA of the blubber oils were mainly n-3 polyunsaturated fatty acids--23:5n-3, 24:3n-3, 24:4n-3, 24:5n-3, 24:6n-3, 26:5n 3, 26:6n-3, and 28:7n-3. The largest VLCPUFA components in all populations were 24:5n-3 (0.1-0.2 wt% of total fatty acids) and 24:6n-3 (0.1%), but 24:4n-3 (0.1%) was also prominent in the Baltic specimens. The blubber oils of the freshwater species contained considerably more 24:4n-6 and 24:5n-6 than the blubbers of the marine species. The differences among the VLCPUFA in these subspecies appear to be mainly due to different dietary VLCPUFA. PMID- 7475989 TI - Cucumber cotyledon lipoxygenase oxygenizes trilinolein at the lipid/water interface. AB - The reactivity of cucumber cotyledon lipoxygenase with trilinolein was examined. The activity of the enzyme against linoleic acid rapidly decreased with increasing pH of the assay solution, and essentially no activity could be detected above pH 8.5. The rapid decrease in activity was not the result of an inactiveness of the enzyme at alkaline pH, because with trilinolein, the enzyme showed a broad pH-activity profile, and substantial activity could be detected even at pH 9.0. Rather, the decrease in activity was due to the dissociation of the linoleic acid emulsion into acid-soap aggregates and/or the monomeric form, depending on the ionization of the terminal carboxylic group. This suggests that cucumber cotyledon lipoxygenase acts only on an insoluble substrate at the lipid/water interface but not on a soluble one. High-performance liquid chromatography analyses of the products formed from trilinolein revealed that the enzyme inserted oxygen into the acyl moiety of trilinolein without hydrolysis of the ester bonds. Preincubation of the enzyme with triolein emulsions effectively abolished its activity against trilinolein added afterward. Furthermore, the enzyme was adsorbed on the trilinolein or triolein emulsion droplets in an essentially irreversible manner. A reaction velocity curve of the enzyme with trilinolein showed saturation kinetics. This is thought to be due to a regional substrate deficiency as the reaction proceeds. These lines of evidence indicate that the enzyme, once bound to the lipid/water interface, is unable to break free and bind to other emulsions. PMID- 7475990 TI - Critical analysis of phospholipid hydrolyzing activities in ripening tomato fruits. Study by spectrofluorimetry and high-performance liquid chromatography. AB - Using the spectrofluorimetric method described by Wittenauer et al. [Wittenauer, L.A., Shirai, K., Jackson, R.L., and Johnson, J.D. (1984) Biochem. Biophys. Res. Commun. 118, 894-901] for phospholipase A2 (PLA2) measurement, we have detected a phospholipase activity in Ailsa Craig and in mutant rin tomatoes at their normal harvest time (mature green stage). This activity in Ailsa Craig tomatoes increased at the beginning of fruit ripening (green-orange stage) and then decreased slowly. The decrease in activity, however, was greater when ripening occurred after tomato picking at normal harvest time than when ripening occurred on tomato plants. This phospholipase activity was always higher in rin tomatoes than in normal ones. Thin-layer chromatography of compounds obtained after incubation of tomato extract demonstrated a decrease in the substrate 1-acyl-2 (6[(7-nitro-2,1,3 benzoxadiazol-4-yl)amino]-caproyl)-sn-glycero-3-phosphocholine (C6-NBD-PC), and an increase in one product (NBD-aminohexanoic acid), but failed to detect the second product (1-acyl-sn-glycero-3-phosphocholine). We, therefore, developed a new one-step method for separation and quantification of a mixture of phospholipids and other lipids, using straight-phase-high-performance liquid chromatography with light-scattering detection. This method detected another fatty acid-releasing activity in enzyme extract from green-orange tomatoes. This lipolytic enzyme (or family of enzymes) slowly produced free fatty acids when 1 oleoyl-sn-glycero-3-phosphocholine was added as substrate. The production of fatty acids was stoichiometric and more rapid when 1-oleoyl-sn-glycero-3 phosphate and 1-oleoyl-sn-glycerol were used as substrates.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7475991 TI - Substrate specificity of the lipase from Candida parapsilosis. AB - Substrate specificity of the acyltransferase activity of the lipase (EC 3.1.1.3) from Candida parapsilosis CBS 604 was studied in aqueous media. The specificity toward both acid and alcohol parts of a large number of acylglycerols and aliphatic esters was investigated. This lipase showed a high activity in the presence of esters with long-chain fatty acids and particularly unsaturated fatty acids with a cis-delta 9 double bond. It was observed that the activity profile depended not only on the alcohol part of the acyl ester, but also on the temperature of the reactant medium. The best lipid substrates had their melting point between -40 to +20 degrees C, 14 to 18 carbon atoms in the acyl group and 1 to 4 carbon atoms in the alkyl group. The enzyme, defined as an acyltransferase in a previous paper, showed a high affinity for primary and secondary alcohols with a short carbon chain (1 to 5 carbon atoms) as acyl acceptors. The influence of free alcohols in the reactant medium on the hydrolysis and alcoholysis activities of the enzyme is discussed. Two phenomena seem to be involved, depending on the alcohol: competition with water for the acyltransfer reaction and lipid substrate dilution when the alcohol places at the oil/water interface. PMID- 7475992 TI - Octadecatrienoic acids as the substrates for the key enzymes in glycerolipid biosynthesis and fatty acid oxidation in rat liver. AB - The activities of key enzymes in glycerolipid biosynthesis and fatty acid oxidation were compared using CoA esters of naturally occurring positional isomers of octadecatrienoic acids (18:3) as the substrates. The trienoic acids employed were 9,12,15-18:3 (alpha-18:3), 6,9,12-18:3 (gamma-18:3), and 5,9,12 18:3 (pinolenic acid which is a fatty acid contained in pine seed oil, po-18:3). The activities of microsomal glycerol 3-phosphate acyltransferase obtained with various 18:3 were only slightly lower than or comparable with those obtained with palmitic (16:0), oleic (18:1), and linoleic (18:2) acids. Mitochondrial glycerol 3-phosphate acyltransferase was exclusively specific for saturated fatty acyl CoA. The activities of microsomal diacylglycerol acyltransferase measured with various polyunsaturated fatty acyl-CoAs were significantly lower than those obtained with 16:0- and 18:1-CoAs. Among the polyunsaturated fatty acids, gamma 18:3 gave the distinctly low activity. The Vmax values of the mitochondrial carnitine palmitoyltransferase I were significantly higher with alpha-18:3 and po 18:3 but not gamma-18:3, than with 16:0 and 18:2, while the apparent Km values were the same irrespective of the types of acyl-CoA used except for the distinctly low value obtained with gamma-18:3. The response to an inhibitor of the acyltransferase reaction, malonyl-CoA, was appreciably exaggerated with 18:2, alpha-18:3, and po-18:3 more than with 16:0 and 18:1. However, the response with gamma-18:3 was the same as with 16:0. Thus, some of glycerolipid biosynthesis and fatty acid oxidation enzymes could discriminate not only the differences in the degree of unsaturation of fatty acids but also the positional distribution of double bond among the naturally occurring 18:3 acids. PMID- 7475993 TI - Inhibition of neutral cholesteryl ester hydrolase by the glycolytic enzyme enolase. Is this a secondary function of enolase? AB - There is an accumulation of the glycolytic enzyme enolase and of cholesteryl esters in macrophages that have been converted into "foam" cells. In this study, we questioned whether enolase could be involved in this accumulation of cholesteryl esters by inhibiting the activity of neutral cholesteryl ester hydrolases. Enolase from both yeast and rabbit muscle were incubated with three different cholesteryl ester hydrolases and were shown to inhibit the hydrolysis of cholesteryl esters. Inhibition was dependent on the concentration of enolase and appeared to occur through binding of the enolase to the cholesteryl ester. Nevertheless, the yeast and rabbit muscle enolases differed in their efficiency of inhibition and in their mechanism of action. Purification of commercial enolase preparations by gel-filtration yielded single proteins with the same inhibitory activities as the originals, indicating that the inhibition was not due to the presence of an impurity. Partially purified alpha alpha- and gamma gamma-isoforms of the enzyme from rat brain also appear to have inhibitory effects on cholesteryl ester hydrolysis. Negative control of the hydrolytic phase of the cholesterol/cholesteryl ester cycle may be a secondary function of enolases which correlates with the accumulation of cholesteryl esters in a number of neuro-degenerative and demyelinating diseases. PMID- 7475994 TI - Inhibition of cholesterol esterification in macrophages and vascular smooth muscle foam cells: evaluation of E5324, an acyl-CoA cholesterol acyltransferase inhibitor. AB - Cholesteryl esters (CE) comprise the principal lipid class that accumulates within macrophages and smooth muscle cells of the atherosclerotic lesion. Acyl CoA cholesterol acyl-transferase (ACAT) is the major enzyme responsible for esterification of intracellular cholesterol. We evaluated the ability of E5324 (n butyl-N'-[-2-[3-(5-ethyl-4-phenyl-1H-imidazol-1-yl)propoxy]-6- methyl phenyl]urea), a novel, orally absorbable ACAT inhibitor, to inhibit esterification of fatty acids to cholesterol and CE accumulation in macrophages and in smooth muscle cells. E5324 significantly inhibited cholesterol esterification in rat aortic smooth muscle cells and in macrophages. In addition, E5324 reduced the cellular mass of CE, the significant measure of the efficacy of drugs designed to modulate cholesterol metabolism. E5324 treatment of macrophages exposed to acetylated low-density lipoprotein reduced CE mass by 97%, and treatment of lipid-loaded smooth muscle cells reduced CE mass by 29%. Although free cholesterol increased approximately twofold, this free cholesterol would presumably be accessible to the membrane for efflux in vivo (reverse cholesterol transport). These results demonstrate that E5324 can inhibit cholesterol esterification and CE mass in atherosclerotic foam cells, derived from either macrophages or arterial smooth muscle cells. PMID- 7475995 TI - Tissue-selective inhibition of sterol synthesis in mice by pravastatin sodium after a single or repeated oral administrations. AB - Pravastatin, an inhibitor of 3-hydroxy-3-methylglutaryl CoA (HMG-CoA) reductase, exhibits liver-selectivity in inhibiting sterol synthesis, when administered as a single oral dose to mice or rats, whereas lovastatin and simvastatin do not. This may be due to the fact that pravastatin is distributed intracellularly, to a large extent, in the liver and extracellularly in nonhepatic tissues. In the present study, we examined whether the difference in liver-selectivity among these three HMG-CoA reductase inhibitors observed in single-dose studies was preserved after repeated oral administrations of drugs to mice. De novo sterol synthesis in different tissues of mice was examined in vivo three hours after the last dose of drug by measuring incorporation of intraperitoneally injected [14C]acetate into total sterols. Pravastatin administered orally for 11 consecutive days at 5 and 10 mg/kg exhibited a greater liver-selectivity than lovastatin and simvastatin: sterol synthesis was inhibited more than 60% in the liver by all three drugs, whereas that in nonhepatic tissues was inhibited less than 10% by pravastatin and more than 30% by lovastatin and simvastatinin in most of the nonhepatic tissues examined. Pravastatin administered orally for 11 consecutive days at 10 mg/kg caused more selective inhibition of sterol synthesis in liver ex vivo than two other inhibitors at the same dose. Pravastatin inhibited de novo sterol synthesis from [14C]acetate into sterol fraction in the liver slices in vitro, but minimally in those of the spleen and testis, whereas lovastatin and simvastatin inhibited in those of all three tissues.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7475996 TI - Synthesis of linoleate and alpha-linolenate by chain elongation in the rat. AB - The objective was to determine whether rats could synthesize longer chain polyunsaturates from hexadecadienoate (16:2n-6) and hexadecatrienoate (16:3n-3). Rats were gavaged with uniformly 13C-labelled hexadecadienoate or hexadecatrienoate, euthanized 24 h later, and total lipids were extracted from liver and carcass. Gas chromatography/combustion/isotope ratio mass spectrometry was used to measure 13C levels in individual liver, carcass, and whole body fatty acids. 13C Enrichment was present in desaturated and chain-elongated polyunsaturates, including linoleate, arachidonate, alpha-linolenate, and docosahexaenoate at 12-13% of the dose of tracer given. 13C Enrichment from hexadecatrienoate was highest in carcass and liver alpha-linolenate, representing 3.5 and 17.9% of the total alpha-linolenate pool, respectively. For linoleate, arachidonate, or docosahexaenoate, the contribution of 13C did not exceed 0.2% of the total body pool. Green leafy vegetables common in the human diet were shown to contain up to 1.2% of total fatty acids as hexadecadienoate and 11.6% as hexadecatrienoate. Hence, humans consuming green vegetables probably synthesize a small proportion of their total body content of linoleate and alpha-linolenate. PMID- 7475997 TI - Cautious optimism: will we ever see a tobacco-free generation? PMID- 7475998 TI - Marketing adolescent tobacco addiction. PMID- 7475999 TI - Involuntary smoking and the health of children. PMID- 7476000 TI - Smoking cessation counseling in adults and children: the clinician's role. AB - Clinicians can profoundly improve the nation's health by implementing recently developed smoking cessation counseling strategies. Although not specifically validated for children and adolescents, young people are likely to benefit from many of the methods used successfully with adults. In the "minimal contact" setting of an office visit, clinicians can achieve substantial long-term quit rates by identifying all smokers through the medical chart, enlisting the help of ancillary personnel, focusing on patients who are ready to quit, and arranging follow-up. Physicians are more likely to be effective if they recognize that smoking cessation is a long-term process that may require multiple quit attempts for success. This article reviews office-based approaches to smoking cessation counseling, tobacco use prevention in children and adolescents, and pharmacologic treatment of nicotine addiction. PMID- 7476001 TI - Developing provider partnerships in the detection of oral cancer and the prevention of smokeless tobacco use. PMID- 7476002 TI - Preventing youth access to tobacco products in Maryland. PMID- 7476003 TI - Tobacco excise taxes: stop children from starting! PMID- 7476004 TI - Maryland restricts workplace smoking. PMID- 7476005 TI - Limiting liability to avoid malpractice litigation. PMID- 7476006 TI - Can breastfeeding be taught in Khmer? PMID- 7476007 TI - In re Baby K. PMID- 7476008 TI - Helping survivors of sexual abuse through labor. PMID- 7476009 TI - Using case management to improve care delivery in the NICU. PMID- 7476010 TI - Acute organophosphate pesticide poisoning in children. PMID- 7476011 TI - A male perspective on infertility. PMID- 7476012 TI - Helping postpartum rural adolescents visualize future goals. PMID- 7476013 TI - Developing hospital-wide standards for i.v. conscious sedation. PMID- 7476014 TI - Ethical perspectives on research involving healthy children. PMID- 7476015 TI - Treating disseminated intravascular coagulation. PMID- 7476016 TI - Cloning of a retinoic acid-induced gene, GT1, in the embryonal carcinoma cell line P19: neuron-specific expression in the mouse brain. AB - Mouse P19 embryonal carcinoma cells can be reproducibly differentiated into neurons and glial cells upon treatment with high concentrations of retinoic acid (RA). To understand the molecular mechanisms that control early neural differentiation, we constructed P19 cell lines carrying an insertion of a gene trap vector containing lacZ as the reporter gene and a G418 resistance gene. We tested expression of the lacZ gene during the RA-induced differentiation process of 300 clones selected with G418. Ten of these clones were stained with X-gal, and five of these ten clones showed up- or down-regulation of lacZ expression. We analyzed one clone, GT1, in which expression of the lacZ gene was markedly up regulated. The 5'-flanking genomic DNA of the GT1 gene present at the site of integration was isolated by the plasmid rescue method, and we screened a cDNA library using this DNA gene as a probe. The GT1 cDNA is about 9000 bp long, with an open reading frame encoding 1840 amino acids. This amino acid sequence has a potential glycosaminoglycan attachment site (Ser-Gly-Gly-Gly) and three N-linked glycosylation sites, but no signal peptide. The sequence of GT1 does not show significant homology with any other known proteins, suggesting that GT1 may be a novel proteoglycan core protein. In situ hybridization revealed that GT1 mRNA was expressed ubiquitiously in the adult mouse brain. This expression was specifically localized in neurons but not in glial cells. Immunohistochemistry revealed that GT1 protein was also localized in neurons. These results suggest that this protein may play a fundamental role in neurons. PMID- 7476017 TI - Molecular cloning of a phosphoinositide-specific phospholipase C from catfish olfactory rosettes. AB - A 2.7 kb clone encoding the partial (about 66%) sequence of a phosphoinositide specific phospholipase C (PLC) was isolated from a cDNA library constructed from channel catfish (Ictalurus punctatus) olfactory rosettes. The clone, designated 30c7, was completely sequenced by automated DNA sequencing and was found to share significant homology with rat and bovine PLCs of the delta 1 isotype. In situ hybridization showed that 30c7 transcripts were expressed in a small subpopulation of olfactory neurons, as well as in other cell types in the olfactory epithelium. Polymerase chain reaction (PCR) analysis indicated that the enzyme was also expressed in several additional tissues, including brain, gill, heart, liver and skeletal muscle. These results suggest that the PLC encoded by clone 30c7 is expressed in several tissues and therefore may have a role in mediating transduction events in diverse tissues as well as in a small group of olfactory neurons. PMID- 7476018 TI - Successive organophosphate inhibition and oxime reactivation reveals distinct responses of recombinant human cholinesterase variants. AB - To explore the molecular basis of the biochemical differences among acetylcholinesterase (AChE), butyrylcholinesterase (BuChE) and their alternative splicing and allelic variants, we investigated the acylation phase of cholinesterase catalysis, using phosphorylation as an analogous reaction. Rate constants for organophosphate (DFP) inactivation, as well as for oxime (PAM) promoted reactivation, were calculated for antibody-immobilized human cholinesterases produced in Xenopus oocytes from natural and site-directed variants of the corresponding DNA constructs. BuChE displayed inactivation and reactivation rates 200- and 25-fold higher than either product of 3'-variable AChE DNAs, consistent with a putative in vivo function for BuChE as a detoxifier that protects AChE from inactivation. Chimeric substitution of active site gorge lining residues in BuChE with the more anionic and aromatic residues of AChE, reduced inactivation 60-fold but reactivation only 4-fold, and the rate-limiting step of its catalysis appeared to be deacylation. In contrast, a positive charge at the acyl-binding site of BuChE decreased inactivation 8-fold and reactivation 30-fold. Finally, substitution of Asp70 by glycine, as in the natural 'atypical' BuChE variant, did not change the inactivation rate yet reduced reactivation 4 fold. Thus, a combination of electrostatic active site charges with aromatic residue differences at the gorge lining can explain the biochemical distinction between AChE and BuChE. Also, gorge-lining residues, including Asp70, appear to affect the deacylation step of catalysis by BuChE. Individuals carrying the 'atypical' BuChE allele may hence be unresponsive to oxime reactivation therapy following organophosphate poisoning. PMID- 7476019 TI - Aberrant expression of neuropeptide Y in hippocampal mossy fibers in the absence of local cell injury following the onset of spike-wave synchronization. AB - Stargazer mutant mice inherit a recessive neuronal excitability phenotype featuring frequent non-convulsive spike-wave seizures that arise from synchronous bursting in neocortical, thalamic and hippocampal networks. Immunocytochemistry reveals that granule cells in the mutant dentate gyrus aberrantly express neuropeptide Y (NPY) at multiple ages following the developmental onset of seizures. The ectopic NPY is selectively concentrated in the mossy fibers, co localizing with the releasable dense core vesicle pool. The NPY content of native NPY+local circuit neurons is also elevated in the mutant CNS. There is no concurrent elevation of hippocampal 72 kDa heat shock protein (HSP72), glial fibrillary acidic protein (GFAP) or NADPH-diaphorase, three markers that are induced during cellular injury, and no evidence of granule cell loss. Since mossy fiber NPY expression appears after the developmental onset of spike-wave discharges and can be induced in wild type granule cells by electrical stimulation, the altered peptide phenotype is likely to reflect transynaptic gene induction triggered by synchronous bursting. These results link a specific pattern of repetitive synaptic input with selective molecular plasticity in dentate granule cells that may contribute to dynamic modifications in hippocampal network excitability. PMID- 7476020 TI - Expression of interleukin 1 alpha, interleukin 1 beta and interleukin 1 receptor antagonist mRNA in mouse brain: regulation by bacterial lipopolysaccharide (LPS) treatment. AB - Lipopolysaccharide (LPS) stimulation is known to induce interleukin-1 (IL-1) mRNA expression in various immune cell types. Since IL-1 synthesis has been suggested to occur locally in brain tissue, we investigated the expression of IL-1 (alpha and beta) and IL-1 receptor antagonist (IL-1ra) mRNAs in various structures of the central nervous system, as well as in the spleen, following intraperitoneal injection of LPS (100 micrograms/mouse). After RNA extraction and amplification by the reverse transcription-polymerase chain reaction (RT-PCR), the PCR products were separated on an agarose gel, transferred and hybridized with digoxigenin labeled probes synthetized by nested PCR. Glyceraldehyde phosphate deshydrogenase mRNA was used as an internal control. Under basal conditions the expression of IL 1 alpha, IL-1 beta and IL-1ra mRNAs in the brain was extremely low for the three cytokines; in the spleen these mRNAs were clearly detectable. Following LPS stimulation, mRNAs were strongly increased in all the tested tissues (cortex, hippocampus, hypothalamus, cerebellum, pituitary and spleen). The kinetics of mRNAs expressions in the brain were similar for all the tested regions, with a maximum at 6 h and a decrease up to 24 h after LPS administration. In the spleen the maximum was observed as soon as 1 h following stimulation. In conclusion, peripheral LPS stimulation induces a strong and transient expression of IL-1 alpha and IL-1 beta mRNAs in the brain. IL-1ra mRNA is also stimulated by LPS in various regions of the brain.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7476021 TI - Biosynthesis of large dense-core vesicles in PC12 cells: effects of depolarization and second messengers on the mRNA levels of their constituents. AB - mRNA levels of various constituents of large dense-core vesicles were determined in PC12 cells during depolarization and/or in the presence of BayK 8644, forskolin or phorbolester. For the soluble (secretory) proteins of the vesicles the mRNAs of chromogranin A and B, secretogranin II, neuropeptide Y and VGF were analyzed. Depolarization in the presence of BayK induced a strong up-regulation of the messages for chromogranin B, neuropeptide Y and VGF. Addition of forskolin enhanced this response for neuropeptide Y and VGF, phorbolester did the same only for VGF. Partly membrane-bound and membrane-spanning components analyzed were carboxypeptidase H, dopamine beta-hydroxylase and glycoprotein III (clusterin), peptidylglycine alpha-amidating mono-oxygenase and cytochrome b-561, respectively. Changes of mRNAs for these components were in general smaller and delayed. Six days of depolarization caused an up-regulation of glycoprotein III, peptidylglycine alpha-amidating mono-oxygenase and carboxypeptidase H mRNA levels which were not further increased by cyclic AMP and phorbolester. The dopamine beta-hydroxylase message increased after 6 days of depolarization, however, addition of phorbolester reduced this effect. For cytochrome b-561 there was no change after any of the conditions employed. These in vitro results are compared with those obtained for the biosynthesis regulation of large dense-core vesicles under in vivo conditions. It is suggested that in vivo acetylcholine and vasoactive intestinal polypeptide released from splanchnic nerve induce a differential change in the biosynthesis of large dense-core vesicles by acting via calcium and protein kinase A and C. PMID- 7476022 TI - N-methyl-D-aspartate and nitric oxide regulate the expression of calcium/calmodulin-dependent kinase II in the hippocampal dentate gyrus. AB - Injection of small volumes of N-methyl-D-aspartate (NMDA) or Sin-1 molsidomine (a nitric oxide releasing agent) onto the dendrites of granule cells in the hippocampal dentate gyrus leads to changes in the level of expression of a number of genes. There is a fall in prodynorphin mRNA levels with a corresponding increase in proenkephalin mRNA levels. Similar changes in opioid gene expression occur following the induction of long-term potentiation (LTP). We report here that at short time periods (1-6 h) after injections of NMDA or sin-1 molsidomine, there is an increase in the levels of the mRNA encoding the alpha subunit of Ca2+/calmodulin-dependent protein kinase II (CaMKII alpha), consistent with a report of elevated CaMKII alpha mRNA in postsynaptic neurons in the CA1 region of the hippocampus following LTP induction [54]. However, we also report that 24 h after injection of NMDA or sin-1, there is a dramatic decrease in CaMKII alpha mRNA levels in the vicinity of the injection. This effect is specific for CaMKII alpha mRNA, in that many other mRNA species are not affected, and occurs in the dendritic population of CaMKII alpha mRNA as well as in the pool of mRNA in the granule cell bodies. The effect is blocked by an inhibitor of cGMP-dependent protein kinase. The biphasic regulation of CaMKII alpha mRNA may be of considerable functional importance for the long-term response of granule cells to local stimulation of NMDA receptors or NO release. PMID- 7476023 TI - N-acetylaspartate as an acetyl source in the nervous system. AB - To understand the role of N-acetylaspartate (NAA) as an acetyl donor, we investigated the metabolism of NAA in brain and liver slice preparations. The tissue slices were incubated with [14C-acetyl]NAA (SA = 3 microCi/mumol) or [14C]acetate (SA = 3 microCi/mumol) for 2 h. The tissue was homogenized and was extracted using chloroform/methanol (2:1). The aqueous phase was initially analyzed using anion exchange HPLC while the lipid phase was analyzed using a two dimensional TLC system. Further resolution of the NAA peak from the anion exchange HPLC was performed using a reverse phase HPLC system. The aqueous phase of both the liver and brain samples incubated with [14C-acetyl]NAA revealed similar patterns of three distinct radioactivity peaks corresponding to NAA, acetate and an early eluting unknown molecule. Further resolution of the NAA peak using reverse phase HPLC indicated that it corresponded to NAA and acetyl CoA. There was significant incorporation of radioactivity into various lipid components in both the brain and liver samples. Patterns similar to that observed with NAA were detected in the case of [14C]acetate in both the brain and liver slice preparations. These results demonstrate that NAA metabolism is not restricted to the nervous system, although its biosynthesis is. It is clear that acetyl moiety of NAA is incorporated into lipids and partially hydrolyzed to free acetate in both brain and liver preparations. Further, production of acetyl CoA from NAA indicates that the acetyl group of NAA is incorporated into lipids and perhaps other acetylated molecules via the acetyl CoA route. A working hypothesis on the metabolic role of NAA is presented. PMID- 7476024 TI - Neurotrophin expression modulated by glucocorticoids and oestrogen in immortalized hippocampal neurons. AB - We have used reverse transcription followed by polymerase chain reaction amplification to investigate changes in expression of nerve growth factor (NGF) mRNA in immortalized hippocampal neurons after treatment with the glucocorticoids dexamethasone and corticosterone, the glucocorticoid antagonist RU38486, and the gonadal steroids progesterone and 17-beta oestradiol. We found that NGF mRNA levels rise after application of either dexamethasone or corticosterone, and that this rise is prevented by the antagonist. Thus, neurotrophin expression is modulated by the physiological glucocorticoid and is mediated by type II glucocorticoid receptors. Progesterone has no effect, while 17-beta oestradiol suppresses NGF mRNA in a postnatally-derived cell line but does not change levels in an embryonic line. An increase in neurotrophin expression is therefore not a general response to steroid hormone application, and may be a specific defence against the presence of metabolically endangering glucocorticoids. PMID- 7476025 TI - Tyrosine kinase phosphorylation of GABAA receptors. AB - Phosphorylation of purified bovine brain GABAA receptors by the tyrosine kinase, pp60v-src was examined. pp60v-src phosphorylated two bands of 54-62 kDa and 48-51 kDa that migrated to approximately the same position as bands recognized by antisera against the beta 2 and gamma 2 GABAA receptor subunits, respectively. Bacterially expressed proteins containing the putative large cytoplasmic loops of the beta 1 and gamma 2L subunits were phosphorylated by pp60v-src, indicating that the phosphorylation sites are located in these subunit domains. The tyrosine kinase inhibitors, genistein and the tyrphostins B-42 and B-44, inhibited muscimol-stimulated 36Cl- uptake in mouse brain membrane vesicles (microsacs). magnitude of the tyrphostin B-44-induced inhibition of muscimol-stimulated 36Cl- uptake was significantly reduced in microsacs that were lysed and resealed under conditions that inhibit phosphorylation. GABA-gated Cl- currents were also inhibited by genistein and tyrphostin B-44 in Xenopus oocytes expressing alpha 1 beta 1 and alpha 1 beta 1 gamma 2L subunits. Consequently, protein tyrosine kinase-dependent phosphorylation appears to be another mechanism of regulating the function of GABAA receptors. PMID- 7476026 TI - Distribution of AMPA-selective glutamate receptor subunits in the human hippocampus and cerebellum. AB - The distribution of AMPA-selective subunits, GluR1-4, was determined in the human hippocampus and cerebellum by in situ hybridization and immunocytochemistry. In the hippocampus, in situ hybridization revealed that GluR1 and GluR2 mRNAs were similarly distributed and highly expressed in the dentate gyrus, with lower levels in the CA regions. GluR3 and GluR4 mRNAs were expressed at very low levels. Immunocytochemical studies showed that GluR1- and GluR2/3 immunoreactivity were highest in the dentate molecular and granular layers. In the CA regions, GluR1 and GluR2/3 staining was observed in pyramidal cell bodies and surrounding neuropil and was more intense in CA4/3/2 compared with CA1. GluR4 immunoreactivity was low throughout the hippocampus. In the cerebellum, GluR1 and GluR4 transcripts were expressed in the granular and Purkinje cell/Bergmann glia layers. GluR2 mRNA was highly expressed in the granular layer and individual Purkinje cells, while GluR3 mRNA was not detectable in the cerebellum. GluR1- and GluR4-immunoreactivity were localized to Purkinje cells and putative Bergmann glia, as well as their processes extending into the molecular layer. GluR2/3 staining was intense in Purkinje cells, with moderate staining in the granular layer. Thus, GluR1-4 subunits are differentially distributed in the hippocampus and cerebellum. In addition, the distribution of subunit mRNA and protein correlate well with each other and with the glutamatergic neuroanatomy of the hippocampus and cerebellum. PMID- 7476027 TI - Differential effects of cysteamine on heat shock protein induction and cytoplasmic granulation in astrocytes and glioma cells. AB - The sulfhydryl agent, cysteamine (CSH), promotes the accumulation of autofluorescent, peroxidase-positive cytoplasmic granules in cultured astroglia akin to those which naturally accumulate in astrocytes of the aging periventricular brain. Both in vitro and in situ, CSH rapidly induces various heat shock proteins (HSP) in astrocytes long before granulation occurs. In the present study, we determined that CSH treatment resulted in an increase in HSP 27, HSP 90 and heme oxygenase (HO-1) at both the protein and mRNA level. We also showed that C6 glioma cells, unlike primary astrocytes, constitutively express HSP 27, HSP 90 and HO-1 at low levels. Moreover, CSH is incapable of eliciting further HSP expression or inducing granulation in the glioma cells. Our results support the hypothesis that the biogenesis of redox-active astrocytic inclusions in CSH-treated glial cultures and in the aging periventricular brain is dependent on an antecedent cellular stress response. PMID- 7476028 TI - Involvement of peroxynitrite in N-methyl-D-aspartate- and sodium nitroprusside induced release of acetylcholine from mouse cerebral cortical neurons. AB - Functional roles of peroxynitrite in N-methyl-D-aspartate (NMDA)- and sodium nitroprusside (SNP)-evoked releases of acetylcholine (ACh) from cerebral cortical neurons in primary culture have been investigated. NMDA increased the release of ACh in a dose-dependent manner, which was significantly suppressed by (+)-5 methyl-10,11-dihydro-5H-dibenzo-[a,d]cycloheptan-5,10-imine (MK-801), a non competitive antagonist specific for the NMDA receptor complex, and NO synthase inhibitors. SNP also showed a concentration-dependent increase in ACh release. Hemoglobin significantly abolished the stimulatory effects of both NMDA and SNP on ACh release. In addition, superoxide anion scavengers such as superoxide dismutase and ceruloplasmin significantly reduced the increased ACh release evoked by NMDA and SNP. Synthesized peroxynitrite dose-dependently elevated the release of ACh. These results indicate that the increased release of ACh by NMDA and SNP is mediated through peroxynitrite formed in the reaction of superoxide anion with nitric oxide produced by NMDA receptor activation and liberated from SNP rather than nitric oxide itself. PMID- 7476029 TI - Bovine and human tau, highly homologous but less crossreactive: implications for Alzheimer disease. AB - Tau is a highly conserved protein during evolution. However, polymerization of tau into paired helical filaments, a characteristic of Alzheimer disease is unique to humans and has not been seen in animals. The cause of this phenomenon is not known. The cDNA-derived amino acid sequences of bovine and human tau are highly homologous. In this study we show that despite the high homology there are marked differences between the immunoreactivities of human and bovine tau. The four antibodies employed in this study were the monoclonal Tau-1 to the bovine protein, two polyclonal anti-bovine tau 92e and 111e and a polyclonal anti-human tau 113e. The monoclonal antibody Tau-1, the epitope of which lies in the amino acid residues 196-215 of bovine tau and residues 189-207 of human tau differing by a single amino acid, a glycine at position 196 in bovine protein with a proline at position 189 in human protein, reacts 2.3 +/- 0.5 times better with bovine tau than with human tau. Both polyclonal antibodies against bovine tau, 92e and 111e react 25 +/- 7 and 45 +/- 17 folds, respectively, better with bovine tau than human tau, whereas the polyclonal antibody 113e to a human tau peptide does not cross-react with bovine tau. The differences in cross-reactivity that are probably due to differences in conformation of tau from the two species were found using both cytosolic extracts and tau isolated from human and bovine brains.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7476030 TI - Transcriptional control of the rat serotonin-2 receptor gene. AB - Previous reports have indicated that, in vivo, the serotonin-2 (5-HT2) receptor is responsive to exogenously administered glucocorticoids. The ability of the glucocorticoid receptor (GR) to influence transcription of the rat 5-HT2 receptor gene was tested in two different experimental paradigms. In both sets of experiments transcription of the 5-HT2 gene was monitored with a promoter reporter plasmid in which the promoter for the 5-HT2 gene was driving the expression of the firefly luciferase gene. In the first, the 5-HT2 promoter reporter plasmid was transfected directly into RS1 cells followed by dexamethasone treatment. In the second set of experiments, the cDNA encoding the GR carried on a separate expression vector was cotransfected into CCL-39 or Neuro 2a cells along with the 5-HT2 promoter-reporter plasmid. These cells were then exposed to dexamethasone. In the RS-1 and CCL-39 transfection experiments, the dexamethasone treatment caused an inhibition of transcription of the 5-HT2 promoter, whereas in the Neuro-2a cells, the dexamethasone treatment stimulated transcription from the 5-HT2 promoter. These responses were dependent on the presence of the GR. The effect of the activated GR would seem to be indirect as sequence analysis of the 4.2 kb preceding the site of transcription initiation revealed only an 11/15 nt match to a putative glucocorticoid response element (GRE), and deletion of this sequence did not alter the response to dexamethasone. Sequence analysis revealed a variety of potential response elements for other known transcription factors, including four potential AP-1 response elements.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7476031 TI - Acute repeated nicotine injections increase enkephalin and decrease AP-1 DNA binding activity in rat adrenal medulla. AB - Previously we reported that a single injection of nicotine decreased AP-1 DNA binding activity in adrenal medullae, although chronic bidaily nicotine (and saline) injections increased this binding activity [15]. Repeated acute nicotine injections (3 mg/kg i.p., 7 injections equi-spaced over a 3 h period) effectively increased adrenal tyrosine hydroxylase [3] and [Met5]enkephalin levels and also profoundly decreased adrenal medulla AP-1 DNA binding activity for over 8 h. PMID- 7476032 TI - Expression of GABAA receptor subunit mRNAs in hippocampal pyramidal and granular neurons in the kindling model of epileptogenesis: an in situ hybridization study. AB - To investigate the molecular changes underlying kindling epileptogenesis in the rat hippocampus, the expression levels of the genes encoding for 13 different gamma-aminobutyric acid type-A receptor (GABAAR) subunits were measured in hippocampal principal neurons using in situ hybridization techniques and semi quantitative analysis of the autoradiograms. Schaffer collateral-commissural pathway kindled rats were investigated at three different stages of kindling acquisition, at 24 h after the last seizure and at long-term (28 days) after termination of kindling stimulations. Changes were distinct for the different subunits in the three analyzed regions (CA1, CA3, fascia dentata) and also different for the various kindling stages. In all hippocampal areas at the early phases of kindling epileptogenesis, before the appearance of generalized seizures, an increase was found of those transcripts that constituted the majority of the expressed variants in control animals (alpha 1, alpha 2, alpha 4, beta 1, beta 2, beta 3, gamma 2/gamma 2L mRNA). In these stages, the increased levels of different variants in the granular neurons of the fascia dentata were more pronounced when compared to the pattern of changes in pyramidal cells of CA1 and CA3. In fully kindled animals, the expression levels of several subunits returned to control levels, whereas beta 3 and gamma 2/gamma 2L mRNA expression was still significantly enhanced in all areas. At long-term, few changes were encountered. The long-splice variant of gamma 2 was decreased within pyramidal and granular neurons while the total level of gamma 2 mRNA was not different from controls. The increased GABAAR subunit expression in the fascia dentata may underly the reported increased GABAAR ligand binding and the increased GABA mediated inhibition. However, the decreased GABAAR binding and the attenuation of GABAergic inhibition in CA1, could not be explained by a decrement of receptor subunit expression. PMID- 7476033 TI - Long-term expression of a reporter gene from latent herpes simplex virus in the rat hippocampus. AB - A problem in utilizing herpes simplex virus (HSV) as a vector for expression of foreign genes in CNS neurons has been the inability to facilitate long-term expression of the engineered genes. Previously, we showed that the murine moloney leukemia virus LTR would drive beta-galactosidase (beta-gal) transcription for extended periods from the latent viral genome in sensory, but not motor neurons. In this communication we further evaluate the utility of the LTR promoter for use in long-term expression vectors. Following stereotactic injection of 8117/43 (an ICP4 minus, non-replicating virus with the LTR driving the beta-gal gene, or KD6 (an ICP4 minus non-replicating virus not expressing beta-gal) into the hippocampus of rats, polymerase chain reaction (PCR) analysis of viral DNA after 2 months indicated that latent infections were established. Assaying by both x gal staining and reverse transcriptase PCR we demonstrate that (1) beta-gal can be detected for at least 6 months in hippocampal neurons, and (2) although the number of beta-gal transcripts in these cells drops considerably by 2 weeks, they can be detected during the period studied. These studies indicate that the LTR promoter is active and affords long-term expression in the CNS, albeit at comparatively low levels compared to those observed at acute times. PMID- 7476034 TI - Spatial and temporal regulation of the rat calmodulin gene III directed by a 877 base promoter and 103-base leader segment in the mature and embryonal central nervous system of transgenic mice. AB - Three non-allelic rat calmodulin (CaM) genes CaMI, CaMII and CaMIII, which share no homology in their 5'-upstream regions, are coordinately expressed in neurons of the central nervous system (CNS). Deletion analysis of the CaMIII promoter showed that the upstream segments longer than 700 bases functioned as efficient promoters, and that the sequence from -133 to -65 was required for the activity of house-keeping type promoter in transient expression assays on a mouse glioma cell line C6. However, the transient expression seemed not to be cell type specific. To determine the temporal and spatial specificity of the promoter function, we produced transgenic mice carrying a fusion gene of the CaMIII segment from -877 to +103 and the lacZ reporter gene. In CNS of the adult transgenic mice, the localization of transgene expression was similar to that of endogenous CaMIII transcripts analyzed by in situ hybridization. The transgene was expressed prominently in pyramidal cells of the cerebral neocortex and the hippocampal regions CA1 to CA3, in Purkinje cells of the cerebellar cortex, and in neurons of the spinal cord, and moderately in granule cells of the dentate gyrus and the cerebellar cortex. In the developing CNS, the overall profiles of neuron-specific expression were also similar for both transgene and endogenous CaMIII that were expressed in the mantle layer and the dorsal root ganglia of the embryonal spinal cord. These results indicated that the neuron-specific expression of rat CaMIII was directed by this 877-base promoter sequence. The CaMIII segment used for the promoter of transgene contained a 29-bp sequence at 410, namely H3, which was conserved in the upstream regions of vertebrate CaMII and CaMIII. H3 seemed to play a pivotal role in the temporal and spatial expression of transgene in CNS, although the deletion of H3 did not decrease CAT activity in the transient expression. The transgene expression was not observed in the external granular cells of the developing cerebellum and in some neurons of the embryonic sensory ganglia in which the endogenous CaMIII was obviously expressed. Therefore, the other cis-acting element(s) located outside of this 877 bp segment seemed to be required for the temporal regulation of CaMIII in certain rudimentary neurons. PMID- 7476035 TI - Limbic effects of repeated electroconvulsive stimulation on neuropeptide Y and somatostatin mRNA expression in the rat brain. AB - The aim of this study was to determine the effect of repeated electroconvulsive stimulation (ECS) on the expression of neuropeptide Y (NPY) and somatostatin (SS) mRNA in the rat brain. For that purpose, quantitative in situ hybridization histochemistry and RNA blot analysis were used. In the hippocampal formation the prevalence of NPY mRNA positive neurons increased in the hilus of the dentate gyrus and the CA3 while a decrease was seen in layers II-III of the entorhinal cortex. In contrast, SS mRNA was increased in the granule cells of the dentate gyrus and in most neurons of the outer parts of the layer III in the entorhinal cortex with cell bodies of perforant pathway projections to the hippocampal CA1 region. Both NPY and SS mRNA expressing neurons were increased in numerical density in the prefrontal cortex with similar amounts of mRNA in individual NPY positive neurons after the stimulations while SS mRNA levels decreased in hybridization positive neurons. In the striatum the only observed significant effect was an increased prevalence of NPY mRNA positive neurons in the caudal nucleus accumbens. Our results provide an outline of a complex functional anatomy of ECS in the rat brain. This type of investigations contributes to map the neuronal systems involved in the action of ECT used in the treatment of affective and schizophrenic disorders. PMID- 7476037 TI - Transient expression of FGF receptor-4 mRNA in the rat cerebellum during postnatal development. AB - The fibroblast growth factor (FGF) receptor-4 mRNA in the adult rat brain is expressed preferentially in the medial habenular nucleus. In this paper, we examined the expression of FGFR-4 mRNA in the brain during postnatal development. Interestingly, in addition to the persistent expression of FGFR-4 mRNA in the medial habenular nucleus, FGFR-4 mRNA was transiently expressed in the proliferative zone of the external granule layer of the developing cerebellum. The localization and transient expression of FGFR-4 mRNA in the developing cerebellum suggest that FGFR-4 mRNA was expressed by proliferative granule cells. The present findings indicate that FGFR-4 in the brain has an important role in the postnatal development of the cerebellar cortex. PMID- 7476036 TI - Distribution of dopamine D1, D2, and D5 receptor mRNAs in the monkey brain: ribonuclease protection assay analysis. AB - The distribution of the mRNAs encoding the dopamine D1, D2 and D5 receptors was determined in brain tissues obtained from intact female rhesus monkeys, using a ribonuclease protection assay. Tissue blocks from the frontal cortex, striatum, thalamus, hippocampus and substantia nigra were dissected and total RNA was extracted. Dopamine D2 and D5 receptor DNA fragments were generated from rhesus monkey genomic DNA using polymerase chain reaction. To generate dopamine receptor subtype-specific cRNA probes, DNA fragments corresponding to the carboxy terminus of the rhesus monkey D1 and D2 receptor genes and to the putative transmembrane domain regions (IV-VI) of the D5 receptor gene, were subcloned into the pGEM3Z/4Z vectors. Expression of D1 receptor mRNA exhibited significant regional differences: striatum > > > cerebral cortex > or = hippocampus > or = lateral thalamus. D1 receptor mRNA was found in low quantities in the medial thalamus, but was not consistently expressed in the substantia nigra area. In contrast, D2 receptor mRNA was detected in all regions that were studied: striatum > > > substantia nigra > > hippocampus > or = cerebral cortex > or = medial thalamus > or = lateral thalamus. D5 receptor mRNA was also expressed in all regions, with highest levels in the cerebral cortex, striatum and lateral thalamus, and moderate levels in the substantia nigra, medial thalamus and the hippocampus. The D5 receptor mRNA appears to be widely distributed in the monkey brain. Most interesting is the expression of D5 receptor mRNA in tissues of the substantia nigra area. PMID- 7476038 TI - [Problems of designing cardiac monitoring systems]. AB - The paper considers one of the ways of solving the problems arising from the design of up-to-date monitoring systems. It describes a Karmon 01M multifunctional fixed monitor manufactured by SEM (Systems of Electronics and Medicine), which may include as many as three randomly chosen modules and replace software for new versions. PMID- 7476039 TI - [Equipment for blood detoxication in critical care of patients]. AB - The paper deals with the development of a new more improved blood detoxicating system in critical patients. Based on the design of a [symbol: see text]-0.5 plasmapheresis device, the authors have designed a new [symbol: see text] 2-0.5 system, by including into the extracorporeal circuit the second rotor which cultures biologically active agents (hepatocytes, fragments of splenic and placental tissue) enhancing a patient's detoxification. The paper also outlines the operating principles of the [symbol: see text] K 2-0.5 device, its operational preparation and procedure. PMID- 7476040 TI - [Comparison of different ways of measuring the obtained dose of hemodialysis]. AB - The paper analyses the reasons for different variations from the calculation of KT/V(ur) by using 9 mathematical models. Analysing the survey data on 120 randomly selected patients on chronic hemodialysis has led to the conclusion that the data obtained by models 1, 3, and 5 more greatly depend on the patient's body compartmentization during a dialysis session than those by models 2, 8, 9 and they generally lower KT/V calculations as they overestimate the urea distribution volume, while with models 2, 8, and 9 the resultant dialysis dose is calculated with less errors. PMID- 7476041 TI - [Prognostic significance of the results of measuring skin capillary permeability in the terminal stage of lower limb ischemia]. AB - The paper outlines a procedure for evaluating distal extremity microcirculation, which consists in exposing the examined skin area to negative pressure and in recording the electric resistance of the area in question prior to and after exposure, the permeability of skin capillaries is judged from the value differences. Its predictive value was assessed in patients with terminal lower extremity arterial ischemia. PMID- 7476042 TI - [Choice of blood pressure determination method in monitoring systems]. PMID- 7476043 TI - [Designing a dynamic spinal distractor]. AB - Based on theoretical calculations, biomechanical tests, and clinical trials, new types of dynamic distractors are proposed to treat scoliosis in children. The specific feature of the designed devices is that they can transform spinal rotatory motions of the spine and move apart the reference elements of the distractor, which rules out the need for supplementary surgical interventions and reduces the incidence of complications. PMID- 7476044 TI - [Magnetocardiographic studies in the clinical diagnosis of heart diseases]. AB - The paper provides evidence for the diagnostic value of magnetocardiography employed in the clinical diagnosis of heart diseases. It concludes that it may be used in mass surveys of the general population. PMID- 7476045 TI - [A comprehensive approach to designing of magnetotherapy techniques based on the Atos device]. AB - The paper determines how to apply a comprehensive approach to designing magnetic therapeutical techniques based on concomitant exposures to two or more physical factors. It shows the advantages of the running pattern of a magnetic field and photostimuli in terms of optimization of physiotherapeutical exposures. An Atos apparatus with an Amblio-1 attachment is used as an example to demonstrate how to apply the comprehensive approach for ophthalmology. PMID- 7476046 TI - [Profile of surfaces of the clamping devices in mechanical medical instruments]. PMID- 7476047 TI - [Automated information system in the diagnostic center]. AB - The paper deals with the problem in automatic medical information control and processing in the diagnostic center. It provides strong evidence for the hard- and software, as well as the pattern of entering data to be stored in the database. It also describes how the system is employed in the Omsk diagnostic center. PMID- 7476048 TI - [Tissue and cell biosensors (possibilities of clinical use)]. AB - The paper provides a review of the data available in the literature on biosensors based on plant and animal tissues. It presents the enzymes contained in the tissues, by describing the respective biochemical reactions underlying these sensors operation. It also gives basic characteristics of the biosensors in question. PMID- 7476049 TI - [Prismatic ophthalmic compensators]. PMID- 7476050 TI - [Neural network algorithms for recognition of complex binary images without their previous segmentation]. AB - The authors propose a new neural network algorithm for binary images. It is based on a binary version of the Grossberg ART-architecture. Its changes made in the mechanism responsible for the competition of categorical neurones are rolled on the neurophysiological findings indicative of the fact that there are intercalary neurons in the inhibitory feedback chains. The proposed scheme in making the competition enables one to supplement the ability to decompose a complex input pattern into the basis (which is frequently non-orthogonal) of previously stored images to the advantages of ART-architectures. PMID- 7476051 TI - [Methods of topographic presentation of the bioelectric activity of the brain]. AB - Prerequisites for developing brain activity topographic mapping and its present status are given. Its advantages and disadvantages, use potentialities in the diagnostic practice of neurological and mental hospitals are discussed. PMID- 7476052 TI - Effects of long-term moderate exercise on iron status in young women. AB - The impact of long-term (6-month) moderate exercise on the iron status of previously sedentary women was determined by randomly assigning 62 college-age women into one of the following four groups: 1) 50 mg.d-1 iron supplement, low iron diet (N = 16); 2) Placebo, free choice diet (N = 13); 3) Meat supplement to achieve 15 mg.d-1 iron intake (N = 13); and 4) Control, free choice diet (N = 20). All groups except the Control group exercised 3 d.wk-1 at 60%-75% of their heart rate reserve. VO2max was measured at baseline and week 24. Blood was sampled at baseline and every 4 wk thereafter for 24 wk to measure iron status and to elucidate the causes for alterations in iron status. Subjects had depleted iron stores throughout the study as indicated by their serum ferritin levels (< 15 ng.ml-1). Serum iron, total iron binding capacity and transferrin saturation were not compromised with exercise. Mean hemoglobin level in the Placebo/Ex group was significantly (P < 0.05) lower than the 50 Fe/Ex and the Meat/Ex groups by week 24. However, changes in serum albumin, haptoglobin, and erythropoietin data from the study cannot explain these changes. PMID- 7476053 TI - Immune response to changes in training intensity and volume in runners. AB - We examined the acute and chronic effects of changes in training volume and intensity on the blood lymphocyte percentages and immunoglobulin levels in runners. Twelve runners participated in four 10-d phases of low volume/low intensity (LV/LI), high volume/low intensity (HV/LI), or high volume/high intensity (HV/HI) running. Subjects were assigned to one of two different training group orders: 1) LV/LI, HV/LI, LV/LI, HV/HI; or 2) LV/LI, HV/HI, LV/LI, HV/LI. Venous blood was drawn at rest on days 1, 4, and 7; and 5 min post exercise on days 1 and 7 of each 10-d phase. Lymphocyte subsets were determined by flow cytometry for CD3+, CD4+, CD8+, and HLA-DR+. IgG, and IgM levels were obtained by ELISA analysis. Immunoglobulin, CD8+ and HLA-DR+ levels, and pre exercise plasma cortisol concentrations were not significantly affected by alterations in volume or intensity. A transient decrease (P < 0.05) was observed in CD4+ and the CD4/CD8 ratio 5 min post-exercise during the HV/LI and HV/HI phases. Results indicate that the exercise-induced lymphocyte subset reduction is transient and suggest that it is more dependent upon training intensity than volume, and the training order of exposure to the high-intensity stimulus may determine the magnitude of subsequent responses. PMID- 7476054 TI - Clenbuterol: a substitute for anabolic steroids? AB - Clenbuterol is a recently popular drug used by athletes in many sports for its purported anabolic effects and reduction of subcutaneous fat. It is a beta-2 (beta 2) agonist prescribed overseas as a bronchodilator, but not approved for use in this country. It is on the banned substance list of the United States Olympic Committee. To avoid any erosion of confidence, physicians caring for athletes need accurate information regarding clenbuterol. Such information is unavailable within the routine medical environs. A review of the literature of animal husbandry reveals that this drug, when administered in doses far greater than those required for bronchodilation, does indeed increase the deposition rate of lean mass and retard adipose gain. There are no human studies available. Animal studies were conducted on laboratory and slaughter stock. No investigation into long-term cardiovascular side effects has been undertaken. The rate of extrapolation from animal studies to unsupervised human usage is alarming. If this category of drugs does preserve lean mass in humans, there are legitimate medical applications. Trials of efficacy and safety are needed. PMID- 7476055 TI - Endothelial-mediated control of coronary and skeletal muscle blood flow during exercise: introduction. AB - There is extensive vasodilation of arterial vasculature in heart and active skeletal muscle during exercise. Considerable knowledge concerning mediators of this vasodilation which act directly on vascular smooth muscle has been acquired. Less well-understood is the role that mediators that act via vascular endothelium may play in exercise-induced vasodilation in vivo. Nonetheless, many studies have been conducted, both on blood vessels in vitro and in resting animals in vivo, since the discovery of endothelial-derived relaxing factor (EDRF) in 1980. In fact, several EDRF have been defined, of which nitric oxide (NO) appears to be most important. Release of NO from vascular endothelium can be induced by two general classes of stimuli, pharmacologic and physical. Of the former class, a physiologically relevant example is norepinephrine. In the latter class, elevated shear stress due to increase blood flow associated with exercise could be a stimulus for NO release. This symposium examines the role that endothelium may play in vasodilation of the coronary and skeletal muscle circulations during acute exercise. It also addresses the question of whether exercise training modifies this role of vascular endothelium. PMID- 7476056 TI - Nitric oxide production and NO synthase gene expression contribute to vascular regulation during exercise. AB - Nitric oxide (NO) is a vasodilator produced under normal physiologic conditions primarily by the vascular endothelium lining all blood vessels. The primary stimulus for the production of nitric oxide by the constitutive endothelial nitric oxide synthase (ECNOS, Type II) found in blood vessels is most likely the shear stress, the frictional force, caused by blood flowing through blood vessels. During exercise there is an increase in cardiac output and redistribution of blood flow to increase blood flow in skeletal muscle and in the coronary circulation. These adjustments provide increased oxygen delivery to support aerobic energy production and to sustain the exercise response. NO may be involved in the regulation of vascular tone in exercising skeletal and cardiac muscle by promoting, enhancing the metabolic vasodilation. In addition, the production of NO by capillary endothelium may regulate oxygen consumption by mitochondria through chemical interactions between NO and the iron-sulfur center of these enzymes. Finally, brief exercise training may alter the gene expression for the enzyme, the constitutive endothelial NO synthase, which forms NO and may be part of the vascular adaptation seen after aerobic exercise training. Furthermore, if there is a genetic predisposition to produce NO, as in world class athletes or animals bred to race, NO may contribute to spectacular exercise performance. These three potential roles of NO will be discussed and data presented to support each of these in our review. PMID- 7476057 TI - Endothelium-mediated control of coronary vascular tone after chronic exercise training. AB - The purpose of this paper is to examine the role of endothelium-derived relaxing factors in the control of coronary vascular resistance in conditioned subjects (i.e., after exercise training for a period of time sufficient to complete adaptation processes). Results from studies with exercise trained (EX) dogs, miniature swine, and rats are summarized. Since the relative importance of vascular control mechanisms differ in various segments of the coronary arterial tree, the effects of EX on conduit arteries and the coronary arterial microcirculation are discussed separately. Results indicate that endothelium mediated vasodilator responses are normal in conduit coronary arteries of EX dogs, miniature swine, and rats. It is proposed that endothelium-mediated vasodilation of conduit coronary arteries is enhanced early in the exercise adaptive process but returns to normal as adaptation to EX is complete, when structural adaptations produce a relative decrease in coronary shear during exercise. EX miniature swine manifest enhanced endothelium-mediated vasodilation stimulated by bradykinin and flow in isolated coronary resistance arteries and appear to have increased expression of NO synthase (ecNOS). Brief training also appears to increase the expression of ecNOS. The role of endothelium-mediated vasodilation in regulation of coronary blood flow in EX animals remains uncertain. PMID- 7476058 TI - Contribution of endothelium-derived nitric oxide (EDNO) to the skeletal muscle blood flow response to exercise. AB - Blood flow (BF) to active muscle increases dramatically during exercise. This increase in BF is permitted by relaxation of smooth muscle (and ensuing vasodilation) in the vasculature of muscle tissue. Recently, attention has focused on the possible role of the endothelium-derived relaxing factor nitric oxide (EDNO) in the vasodilation of muscle vasculature during exercise. A variety of experimental approaches have been used in elucidating the role of EDNO. These include isolated vessel, isolated muscle or muscle group, and conscious exercising animal preparations. Studies utilizing isolated vessels have shown that arterioles from muscle dilate, in an endothelium-dependent manner, to stimuli present during exercise (e.g., increased flow rates). A limitation of such studies, however, is that only the potential for EDNO-induced vasodilation is indicated. The isolated muscle/muscle group approach has consistently demonstrated a role for EDNO in determining resting BF. Findings for muscle BF during contractions are equivocal. A limitation of this approach is that exercise is simulated by stimulating the motor neuron of the muscle of interest. Since this type of muscle activity elicits a relatively small active hyperemia, it may be that a role for EDNO in exercise-induced hyperemia is masked. Findings from exercising animals are equivocal. Some studies demonstrate a role for EDNO in permitting increased muscle blood flow during exercise, while others show no impact of inhibition of EDNO synthesis. Some studies suggest that the importance of EDNO may vary with the muscle (and its fiber type composition) studied. Additional research is needed to clarify the role of EDNO in mediating increased BF to skeletal muscle during exercise. PMID- 7476059 TI - Effects of exercise training on endothelium-dependent peripheral vascular responsiveness. AB - Endurance training results in peripheral vascular adaptations in skeletal muscle which enhance perfusion and vascular flow capacity. These adaptations could result from structural modifications of the vasculature and/or alterations in the control of vascular tone. One potential mechanism through which vascular control may be modified is through adaptive changes in the intrinsic responsiveness of vascular endothelium. Experiments have demonstrated that vascular responsiveness to endothelium-dependent vasodilators are enhanced in exercise-trained animals. The enhanced endothelium-dependent relaxation appears to be mediated through elevations in the formation of endothelium-derived nitric oxide. Training also results in a decreased sensitivity to the vasoconstrictor effects of norepinephrine. This alteration appears to be due to an endothelium-dependent mechanism involving alpha 2-adrenergic receptors. One stimulus that appears to be important in initiating the adaptation of the endothelium to training is the increase in muscle blood flow and shear stress which occurs during exercise. However, other factors associated with exercise may be necessary to induce endothelial adaptations produced by endurance training. Further research is needed to determine the significance of changes in endothelium-dependent vascular responsiveness and whether this is associated with training-induced increases in muscle perfusion and vascular flow capacity. PMID- 7476060 TI - Coordination of blood flow control in the resistance vasculature of skeletal muscle. AB - The coordination of blood flow control in exercising skeletal muscle is exemplified by the interaction among muscle fibers, nerves, and the smooth muscle and endothelial cells which comprise the resistance vasculature. During functional hyperemia in active muscle, maximal flow exceeds resting values by 10- to 50-fold, according to muscle fiber type and recruitment pattern. The control of muscle blood flow is coordinated among many vessel branches, encompassing the resistance arteries external to the muscle and the arteriolar network embedded within the tissue. As motor unit recruitment and metabolic demand increase with exercise intensity, the locus of blood flow control "ascends" from distal arterioles, which govern capillary perfusion and flow distribution, into the proximal arterioles and feed arteries, which control the volume of flow into a muscle. The organization of vasomotor responses within and among resistance vessels can be explained by the spread of electrical, chemical, and physical signals between endothelial and smooth muscle cells. These signals are triggered by substances released from muscle fibers and nerve terminals and by changes in transmural pressure and luminal flow. Thus, from several perspectives, cell-to cell communication coordinates blood flow control in accord with the metabolic demands of active muscle fibers. PMID- 7476061 TI - Endothelial cell calcium and vascular control. AB - The endothelium has been shown to contribute to the modulation of vascular smooth muscle tone. Using digital imaging microscopy, we measured calcium in endothelial cells of isolated skeletal muscle arterioles. First-order arterioles (N = 6) were isolated from rat cremaster muscle and cannulated. All vessels developed spontaneous tone (99.4 microns; 60%-80% of passive diameter). Endothelial cells were loaded with Fura-2-AM (5 microM), a ratiometric calcium-sensitive fluorescent dye. The Fura-2-AM was placed in the lumen of the vessel for approximately 10 min and removed. Fluorescent images of endothelial cells were acquired following excitation at 340- and 380-nm wavelengths and the data expressed as the 340/380 ratio which is directly proportional to intracellular calcium. Acetylcholine (Ach; 10(-6) M), an endothelium-dependent dilator, caused significant dilation (132% of control) and increased calcium to 186% of control. Adenosine (Adn; 10(-4) M), an endothelium-independent agent, produced similar dilation (127% of control) but did not alter endothelial cell calcium. Increasing intraluminal pressure produced a myogenic constriction with no change in endothelial calcium. Flow, induced by a 20 cm H2O pressure gradient, failed to dilate the arterioles and produced no increase in endothelial cell calcium. However, flow induced by a 40 cm H2O pressure difference did increase endothelial cell calcium prior to a significant arteriolar dilation. Removal of the endothelium by physically rubbing the intimal surface eliminated both the dilation and increase in calcium to Ach or flow (40 cm H2O) while the response to Adn remained unchanged.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7476062 TI - Relationship between mean habitual daily energy expenditure and maximal oxygen uptake. AB - A population of 120 healthy voluntary subjects of both genders aged 16-88 was studied using the QAPSE (Saint-Etienne Physical Activity Questionnaire) with the purpose of investigating the factors influencing the relation between MHDEE (mean habitual daily energy expenditure) and VO2max (maximal oxygen uptake) to elucidate the factors accounting for individual variation. The mean of MHDEE obtained was 12,181.9 +/- 4041.9 kJ.d-1. The mean VO2max obtained was 39.9 +/- 13.8 ml.kg-1.min-1. A strong relationship between MHDEE and VO2max (r = 0.916; N = 120; P < 0.0001) was found. Further, MHDEE seemed to be the greater determinant in the variation of VO2max (89.35%). Other variables were found to be involved in the relation between MHDEE and VO2max for a smaller, but still substantial part: age (6.92%), PAST (exathletes who had considerably reduced or stopped their training) (2.45%), body mass (0.85%), and gender (0.43%). Two variables regarding maximal intensity of activity were not included in the multiple-linear regression analysis. These results suggested that the most important factor in the variation of VO2max is the total quantity of energy expenditure and not only the maximal intensity that could reach the subject. PMID- 7476063 TI - The natural history of exercise: a 10-yr follow-up of a cohort of runners. AB - To determine the pattern of exercise and associated adverse events, including injuries, a mail survey was conducted on participants in the 1980 Peachtree Road Race Study for whom we were able to obtain current addresses. A total of 535 persons responded, 326 men and 209 women, representing 72% of those with identifiable addresses. Although only 56% of respondents reported that they were still running in 1990, 81% reported that they were still exercising regularly. The cumulative probability for continuing to run was 0.71 for men and 0.56 for women. Injury (31%) was the chief reason given by men for stopping permanently. For women, 28% stopped because they chose another form of exercise. Fifty-three percent of respondents had at least one injury during the 10-yr interval. The probability of experiencing an injury was associated with higher weekly mileage. The knee was the most frequently injured site. Thirty-nine percent of women and 35% of men reported being verbally assaulted. Approximately 10% of persons were hit by thrown objects or bitten by a dog. In a group of recreational runners, almost half had stopped running 10 yr later, but over 80% were still physically active. Many runners sustain injuries or suffer hazards related to their exercise. PMID- 7476064 TI - Neuromuscular dysfunction following eccentric exercise. AB - This study examined the effects of exercise-induced muscle damage on tremor and proprioception components of neuromuscular function. Six male and six female volunteers (aged 18-30 yr) performed 50 maximal eccentric muscle actions using the forearm flexors of the nondominant arm. Forearm flexor tremor and perception of voluntary force and joint position were monitored to assess changes in neuromuscular function. Data were analyzed using REANOVA. Serum creatine kinase activity increased from a baseline value of 68 +/- 13 IU.l-1 to 2849 +/- 852 IU.l 1 5 d after exercise (P < 0.05). This was accompanied by prolonged impaired joint range of motion (P < 0.01) and reduced maximum strength (P < 0.01). Muscle soreness peaked 3 d postexercise (P < 0.01; Wilcoxon test). Tremor amplitude was increased (P < 0.01) until 48 h after exercise, whereas the power frequency spectrum was unaffected. Perception of joint position at elbow angles of 1.57 rad (P < 0.01) and 2.09 rad (P < 0.05) and perception of force (P < 0.01) were significantly impaired when the control arm acted as the reference. Joint positions were more accurately reproduced when the experimental arm acted as its own reference. The increase in tremor amplitude and loss of proprioceptive function in the days after damage-inducing eccentric exercise suggest significant impairment of neuromuscular function. PMID- 7476065 TI - Effects of stride frequency on mechanical power and energy expenditure of walking. AB - The energetics and mechanics of walking were investigated at different speeds, both at the freely chosen stride frequency (FCSF) and at imposed ones (up to +/- 40% of FCSF). Metabolic energy expenditure was minimized at FCSF for each speed. Motion analysis allowed to calculate: the mechanical internal work rate (Wint), needed to move the segments with respect to the body center of mass (bcm); the external work rate (Wext), necessary to move bcm in the environment; and the total work rate (Wtot), equal to Wint+Wext. Wtot explains the metabolic optimization only at high speeds, while Wext, differently from previously reported, displays minima which better predict FCSF at all speeds (exception made for 1.39 m.s-1). This is probably caused by an overestimation of Wint due to a more ballistic movement of the limbs at low speeds (and low frequencies). The tendency of Wext to increase at high frequencies is due to a persistent minimal vertical excursion of bcm (about 0.02 m, the "locomotory dead space"). While the match between mechanics and energetics (at FCSF and imposed frequencies) occurs to a certain extent, it could be improved by removing the methodological assumptions about the energy transfer between segments and by the possibility to account for the coactivation of antagonist muscles. PMID- 7476066 TI - Influence of high-resistance and high-velocity training on sprint performance. AB - The purpose of this study is to analyze the effect of high-resistance (HR) and high-velocity (HV) training on the different phases of 100-m sprint performance. Two training groups (HR and HV) were compared with two control groups (RUN and PAS). The HR (N = 22) and HV group (N = 21) trained 3 d.wk-1 for 9 wk: two strength training sessions (HR or HV) and one running session. There was a run control group (RUN, N = 12) that also participated in the running sessions (1 d.wk-1) and a passive control group (PAS, N = 11). Running speed over a 100-m sprint was recorded every 2 m. By means of a principal component analysis on all speed variables, three phases were distinguished: initial acceleration (0-10 m), building-up running speed to a maximum (10-36 m), and maintaining maximum speed in the second part of the run (36-100 m). HV training resulted in improved initial acceleration (P < 0.05 compared with RUN, PAS, and HR), a higher maximum speed (P < 0.05 compared with PAS), and a decreased speed endurance (P < 0.05 compared to RUN and PAS). The HV group improved significantly in total 100 m time (P < 0.05 compared with the RUN and PAS groups). The HR program resulted in an improved initial acceleration phase (P < 0.05 compared with PAS). PMID- 7476067 TI - Comparative effectiveness of accommodating and weight resistance training modes. AB - Six men and six women trained the elbow flexors of both arms 3 d.wk-1 for 20 wk. In each training session, one arm did 3-5 sets of 10 maximal concentric actions on an accommodating resistance device (ARD), the other arm 3-5 sets of 8-12 coupled eccentric/concentric actions (repetitions) to volitional failure (8-12 RM) on a weight resistance device (WRD). The average "intensity" (force of concentric actions) was approximately 1.25 times greater in ARD training, the average "volume" (number of actions x force of actions) 1.6 times greater in WRD training, and the time required to complete a training session the same for each. Both types of training produced significant increases in a single maximum weight lift (1 RM on the WRD), in the peak force of a single maximal concentric action measured on the ARD and an isovelocity dynamometer, and in biceps, brachialis, and total elbow flexor cross-sectional area (CSA). Biceps Type I and II fiber area did not change significantly. WRD training produced greater increases than ARD training in the 1 RM test on the WRD and in brachialis CSA. The data indicate that both of these common training regimens effectively increase strength and muscle mass, but the weight training regimen may be more effective for increasing muscle mass. PMID- 7476068 TI - The Wisconsin wrestling minimum weight project: a model for weight control among high school wrestlers. AB - In 1989, the Wisconsin Interscholastic Athletic Association implemented a project including new rules and an educational program, consistent with ACSM and AMA guidelines, to curtail "weight cutting" among high-school wrestlers. The project included skinfold estimates of body fatness to determine a minimum competitive weight, a limit on weekly weight loss, and presentation of nutrition education information to help wrestlers diet effectively. Implementation of the project has overcome a variety of obstacles and has received widespread endorsement from parents, teachers, wrestlers, and coaches. The success of this project offers a model for other states to emulate and should encourage clinicians in other sports to initiate interventions addressing these unhealthy weight loss behaviors. PMID- 7476069 TI - Ongoing initiatives by ACSM on exercise in America. PMID- 7476070 TI - Optimized isotropic diffusion weighting. AB - The authors introduce several sets of time-efficient gradient waveforms for applying isotropic diffusion weighting in NMR experiments. This creates signal attenuation that depends on the trace of the diffusion tensor and is therefore rotationally invariant. Numerical methods for the calculation of such gradient sets are outlined, and results are shown for isotropic and anisotropic gradient hardware and first order flow moment nulled diffusion weighting gradients. Preliminary experimental results from the human brain validate this new technique. PMID- 7476071 TI - 3D echo planar imaging: application to the human head. AB - In this work, the authors present 3D images acquired from the human head using echo planar encoding for two of the three dimensions of k-space. The third dimension of k-space is filled by selecting and phase encoding a slab of spins as in conventional 3D steady state (GRASS based) acquisition regimens. Using this approach, a 128 x 64 x 64 3D data matrix could be obtained in 3.4-4.7 sec using effective TE values of 24 and 34 ms, respectively. High quality 3D images could be acquired once phase ghosts present on 2D images were minimized through proper adjustments of scanner hardware. PMID- 7476072 TI - Increased flexibility in GRASE imaging by k space-banded phase encoding. AB - GRASE (GRadient and spin Echo) is an echo train imaging technique that combines gradient and RF refocusing. Although overall signal decay is with T2 and field inhomogeneity phase errors do not accumulate, the small residual phase errors are periodic with echo number. The echo order described previously eliminates the phase error periodicity in k space but instead creates periodicity in the T2 modulation function that can also cause artifacts. In addition, with this order, the effective TE must be half the echo train time, and asymmetric Fourier sampling is difficult to implement. A new method is described that greatly reduces artifacts due to T2 decay, permits greater control of T2 contrast, and lends itself to asymmetric Fourier sampling. Different time segments of the echo train are encoded with different bands of spatial frequency in k space (hence "k banding"). Both computer simulations and experimental results demonstrate improvements in GRASE images acquired by this method. PMID- 7476073 TI - Optimized gradient waveforms for spiral scanning. AB - Spiral scanning gradient waveforms can be optimized with respect to blurring from off-resonance effects by minimizing the readout time. This is achieved by maximizing the gradient amplitude during the scan so that the edge of k-space is reached as quickly as possible. Gradient hardware constraints are incorporated by considering a circuit model for the gradient coil and amplifier. The optimized gradient waveforms are determined by a set of coupled differential equations. The resulting solutions have shorter readout time than solutions that do not consider the circuit model. PMID- 7476074 TI - Quantitative analysis of regional left ventricular function after myocardial infarction in the pig assessed with cine magnetic resonance imaging. AB - To assess the accuracy of quantitative analysis of global and regional wall motion and wall thickening of the left ventricle with cine magnetic resonance (MR), images obtained in eight pigs before and after myocardial infarction were compared with those obtained using gadolinium diethylenetriaminepentaacetic acid (Gd-DTPA)-enhanced multislice spin-echo MR imaging and determination of pathology. The region with abnormal wall motion and wall thickening, as determined with cine MR imaging, identified the same region of infarction as indicated by Gd-DTPA-enhanced spin-echo MR imaging and pathology. Within the infarcted region wall motion and wall thickening analyzed with the centerline method were significantly reduced. We conclude that the use of quantitative analysis of cine MR images accurately determines localization and extent of regional left ventricular dysfunction in the infarcted heart in vivo. This analysis using dedicated software including the centerline method allows sequential assessment of regional left ventricular function in normal and infarcted hearts. PMID- 7476075 TI - Analytical resolution and noise characteristics of linearly reconstructed magnetic resonance data with arbitrary k-space sampling. AB - The effects of time-varying readout gradients and data sampling with variable dwell times in magnetic resonance imaging are examined. General reconstruction formulas are given for linear reconstruction with even k-space weighting. Closed analytic expressions for estimator variance are given for data sampling during arbitrary gradient waveforms with both uniform kx step size and nonuniform kx step size. It is shown that estimator variance increases (the signal-to-noise ratio decreases) for nonconstant gradient waveforms. It is also shown that estimator variance is greater for constant k-space sampling strategies than for constant time sampling at the Nyquist rate. Data collected during a triangular readout gradient waveform, with either constant time or constant k-space sampling, versus conventional (constant gradient) collection confirms theoretical predictions for estimator variance. The benefits of collecting data while the readout gradient is ramping up from and down to zero are discussed. PMID- 7476076 TI - Abbreviated moment-compensated phase encoding. AB - To achieve correct spatial location of blood vessels, first order gradient moment nulling applied to the phase encoding axes can be used. However, gradient moment nulling prolongs echo time (TE), which may degrade the flow image in regions of complex flow. The fact that abbreviated moment compensated phase-encoding (AMCPE) can be used to apply partial flow compensation to the phase-encoding axes to prevent spatial misregistration of vessels without requiring the use of long echo times or using arbitrary chosen TE is demonstrated. AMCPE defines two cutoff lines in k-space. The flow-induced phase is completely compensated for values between the cutoff lines and partially compensated beyond the cutoff lines. The AMCPE technique has been tested on both a flow phantom and a human volunteer. The AMCPE images from both the in vivo and the in vitro study demonstrate correctly imaged flow. Computer simulations have been performed to analyze the penalty caused by the incomplete flow compensation. The result shows that the ripple artifacts due to the incomplete flow compensation are unobservable when 60%-70% of k-space is completely flow compensated. PMID- 7476077 TI - An improved quadrature or phased-array coil for MR cardiac imaging. AB - A tailored receive-only coil for cardiac imaging has been designed. The coil consists of two overlapping coil elements and can be used either as a quadrature surface coil or as a phased-array coil. Through phantom experiments and images of the heart, the authors have shown that the improved cardiac coil provided a signal-to-noise ratio 1.6 times higher than a conventional quadrature spine coil, 1.4 times higher than that of a single coil (having the same shape and total dimension), and three times higher than the body coil at the depth of the posterior wall of the heart. The authors have also shown that the cardiac coil improved image quality everywhere in the heart. This coil will enhance routine clinical cardiac studies as well as other examinations such as myocardial perfusion, wall motion, and coronary artery imaging. PMID- 7476078 TI - Analytical expressions for the NMR apparent diffusion coefficients in an anisotropic system and a simplified method for determining fiber orientation. AB - NMR measurements of anisotropic diffusion were studied using a three-dimensional random-walk model. It was found that the apparent diffusion coefficient can be expressed in a canonical form as the product of a diagonal matrix, an orthonormal rotation matrix, and a vector representing the encoding magnetic field gradient. The diffusion coefficient can be interpreted as the sum of the corresponding coefficients measured along the principal diffusion axes, weighted by the squares of the directional cosines of the encoding direction with respect to the principal axes. The analysis revealed that determining the orientation of anisotropy, in a cylindrically symmetric system, requires a minimum of four diffusion measurements. A special pulse sequence which minimized gradient cross terms and possible restricted diffusion effects was used to characterize diffusion anisotropy in cut chicken gizzards. Diffusion coefficients parallel to the muscle fibers were found to be approximately two to three times larger than those in the transverse direction. Furthermore, the method was successful in detecting the angular change when the sample was rotated by 30 degrees. Results indicate that the proposed approach to measure fiber orientation is valid and may be used to improve the time efficiency of diffusion anisotropy measurements. PMID- 7476079 TI - Retrospective estimation and correction of physiological fluctuation in functional MRI. AB - Image-to-image fluctuation due to physiological motion is a major limitation to the accurate detection of neuronal activity with functional MRI. In this paper, a new and general technique for the estimation and compensation of the physiological effects is presented. By simultaneously monitoring the respiration and heart beat during the acquisition of imaging data, and retrospectively synchronizing the imaging data with physiological activity, physiological effects are estimated and removed. This technique does not rely on the periodicity of the respiration or the heart beat, does not affect the signal changes arising from neuronal activation, and is beneficial to images acquired with any speed. Experimental studies performed with FLASH and EPI sequences have demonstrated that the new technique is effective in reducing physiological fluctuation and improving the sensitivity of functional MRI and is generally applicable. PMID- 7476080 TI - Modulation of water diffusion during gonadotropin-induced ovulation: NMR microscopy of the ovarian follicle. AB - The preovulatory rat follicle reaches a diameter of 1 mm with no internal blood vessels. Nutrient supply to the enclosed oocyte depends solely on passive diffusion across the follicular wall and the follicular fluid. Spin-echo and stimulated-echo NMR microscopy experiments were applied here for studying modulations in water diffusion during gonadotropin-induced maturation of perfused rat ovarian follicles (32 degrees C). Two diffusion compartments were observed for the follicular wall. The intracellular water diffusion coefficient, measured at a short diffusion time (9 ms) was 0.28 x 10(-5) cm2/s. Diffusion at long diffusion times was restricted to 16 microns, the size of cells in the follicular wall, and did not change during maturation. In the follicular fluid a transient 26% decrease in the diffusion coefficient was observed 4-7 h after gonadotropin stimulation, a change that is bound to affect the metabolic balance of the oocyte before ovulation. PMID- 7476081 TI - Oximetry by dynamic nuclear polarization. AB - This study describes quantitatively the effect of O2 on the dynamic nuclear polarization (DNP) of water protons. It is shown theoretically how DNP enhancement is altered by the paramagnetism of O2. The nonlinear effect of oxygen on the enhancement factor has been approximated by a first order expression that provides accurate predictions of the experimental results obtained with phosphate buffered solutions of 2,2,6,6 tetramethyl-4-oxo-piperidine-N-oxyl. The proton relaxation rates and DNP line width were measured for nitroxide concentrations ranging from 0.5 to 10 mM and for oxygen partial pressures ranging from 0 to 760 mmHg. A method is suggested for the calculation of the nitroxide and oxygen concentrations of a sample, based on two measurements, one with on-resonance EPR irradiation, the other with slightly off-resonance irradiation. Although the correlation between the values for the actual and calculated oxygen concentrations is good, the accuracy of the calculated oxygen concentration values remains poor, because of the large number of parameters involved in this calculation. PMID- 7476082 TI - Theory of 1/T1 and 1/T2 NMRD profiles of solutions of magnetic nanoparticles. AB - Organically coated iron oxide crystallites with diameters of 5-50 nm ("nanoparticles") are potential magnetic resonance imaging contrast agents. 1/T1 and 1/T2 of solvent water protons are increased dramatically by magnetic interactions in the "outer sphere" environment of the nanoparticles; subsequent diffusive mixing distributes this relaxation throughout the solvent. Published theory, valid for the solute magnetic energy small compared with thermal energy, is applicable to small magnetic solutes (e.g., gadolinium and manganese diethylenetriaminopentaacetic acid, and nitroxide free radicals) at generally accessible fields (< or = 50 T). It fails for nanoparticles at fields above approximately 0.05 T, i.e., at most imaging fields. The authors have reformulated outer sphere relaxation theory to incorporate progressive magnetic saturation of solute nanoparticles and, in addition, indicate how to use empirical magnetization data for realistic particles when their magnetic properties are not ideal. It is important to handle the effects of rapid thermally induced reorientation of the magnetization of the nanoparticles (their "superparamagnetism") effectively, including their sensitivity to particle size. The theoretical results are presented as the magnetic field dependence (NMRD profiles) of 1/T1 and 1/T2, normalized to Fe content, for three sizes of particles, and then compared with the limited data extant for well-characterized material. PMID- 7476083 TI - NMR relaxation rates and blood oxygenation level. AB - A linear relationship between the proton relaxation rates and the fraction of deoxyhemoglobin of circulating whole blood (fHb) has been established in vitro at 4.7 T. These results have been interpreted on the basis of the additivity of relaxation rates. The slope of the lines of transverse relaxation rates (R2) versus fHb was found to increase with interpulse delay in Carr-Purcell Meiboom Gill (CPMG) experiments. The Luz-Meiboom relation applied to this interpulse delay dependence of R2 suggests a two-site chemical exchange rather than a diffusion mechanism. The 1-ms water proton exchange time derived from these observations has been interpreted in terms of exchange between exchangeable protons close to the paramagnetic center of hemoglobin and protons of bulk water. PMID- 7476084 TI - In vivo proton MR spectroscopy of human gliomas: definition of metabolic coordinates for multi-dimensional classification. AB - Several multi-dimensional statistical evaluation methods were applied to single voxel proton MR spectra of glial brain tumors and of healthy volunteers. Metabolic coordinates with histological relevance for future diagnosis were found by which spectra from controls, low-grade tumors, and high-grade tumors were completely separated. Significant differences between low-grade and high-grade glioma patients and controls were found for several metabolic ratios by variance analysis. Cluster analysis both with and without principal component analysis was applied. The outcome of these two approaches depended mainly on the lipid-to creatine ratio. Two other approaches, discriminant factor analysis and the orthonormal discriminant vector method were then used to find discriminatory metabolic coordinates. It turned out that a linear combination of all evaluable metabolic ratios made it possible to separate the three groups completely. On the basis of these results, a classification method that uses the entire proton MRS spectrum is proposed. PMID- 7476085 TI - Homonuclear J-refocused spectral editing technique for quantification of glutamine and glutamate by 1H NMR spectroscopy. AB - Quantification of the excitatory amino acid of glutamine and glutamate is extremely important partly because, when in excessive amounts, they can produce neuronal damage. However, quantification of such metabolites seems to be not straightforward owing to a large number of metabolic signals, particularly between the 2.0- and 3.0-ppm chemical shift range. The authors investigated a quantification method by using a homonuclear J-refocused spectral editing technique, namely the J-refocused PRESS (JPRESS), using both 1.5- and 4-T whole body NMR systems. At first, the J-coupling modulation of each metabolite at different TE values using the PRESS and JPRESS pulse sequences was compared. The suppression of J modulations with the JPRESS is well demonstrated on condition that TE values are less than 1/4J. Then, the accuracy of glutamate quantification after subtracting the two spectra obtained at two different echo times and integrating the difference spectral area at different concentrations was evaluated. The results exhibit an excellent linear relationship between the actual concentration versus the difference spectral peak area at both 1.5 and 4 T. The achievable minimum voxel size was also considered and a 1-ml voxel size seems to be possible at the typical glutamate concentration in vivo at a field strength of 4 T. PMID- 7476086 TI - Development of a phased-array coil for the lower extremities. AB - A phased-array coil was developed to facilitate imaging the vasculature of the lower extremities. The array consists of four surface coils placed in a Plexiglas "I-beam" frame that are configured to allow bilateral studies with up to a 40-cm field of view (FOV). Data from phantoms indicate an increase in signal-to-noise ratio (SNR) in the regions of interest by an average factor of 2.8 +/- 0.9 over that of the body coil. Preliminary in vivo data have also been obtained from n = 8 subjects and demonstrate significant improvements in image quality. The coil design described here should lead to reduced scan times through the ability to image both legs simultaneously with less need for patient repositioning. PMID- 7476087 TI - A computer simulation of the static magnetic field distribution in the human head. AB - Distortion of the static magnetic field inside the human head is dependent on regional tissue susceptibility variations and geometrical shape. These effects result in resonance line broadening and frequency shifts and consequently, intensity and spatial errors in both magnetic resonance imaging (MRI) and magnetic resonance (MR) spectroscopy. To calculate the field distortion due to the susceptibility's geometry, two dimensional (2D) finite element analysis was applied to simulate the field distribution in a 2D model of the human head, placed in a uniform magnetic field. The model contains air-filled cavities and sinuses, and the remainder is treated as water. The magnetic field deviation was evaluated using gray scale plots and histograms of the magnetic field. The shifts in parts/million and broadening of the histograms correspond to the NMR of the sampled region. The field distribution of the human head was also experimentally mapped using the DANTE tagging sequence. The calculated and experimental field maps are in good agreement. Thus, geometric considerations with uniform susceptibilities are sufficient to explain most of the static magnetic field distribution in the human head. PMID- 7476089 TI - The application of magnetization transfer to MR angiography with reduced total power. AB - Magnetization transfer (MT) techniques have been shown to significantly reduce background soft-tissue signal in time-of-flight magnetic resonance angiography. To achieve sufficient suppression, radio frequency (RF) pulses with tip angles on the order of 1000 degrees are typically used, resulting in significant RF power deposition in the patient. Although these power deposition levels do not exceed the FDA guidelines, they are significantly higher than those used in typical imaging techniques. The use of these same magnetization transfer pulses in applications at field strengths higher than 1.5 T will require MT power levels which exceed FDA safety standards. This report demonstrates that the total power deposition required to achieve background tissue suppression can be significantly reduced by the application of the saturation pulses only during the phase encoding steps corresponding to the central portion of "k space." This technique allows equivalent soft tissue suppression with approximately 10% of the energy deposition of conventional magnetization transfer techniques. PMID- 7476088 TI - A magnetization-driven gradient echo pulse sequence for the study of myocardial perfusion. AB - A T1-weighted imaging pulse sequence for contrast-based studies of myocardial perfusion is presented and evaluated in phantoms and in vivo. The sequence is similar to spoiled gradient-recalled echo sequences except that nonselective preparatory RF pulses drive magnetization to steady state prior to image acquisition. Steady state is thus obtained in both tissue and blood resulting in a stable, homogeneous, and dark pre-contrast baseline. Tip angles and timings are chosen so that pixel intensity approximates a linear relation to 1/T1. The dynamic range of signal response to contrast agent concentration is greater than that of an inversion-recovery fast low angle shot sequence. The sequence proposed should be useful for myocardial perfusion studies. PMID- 7476090 TI - Purified Pasteurella haemolytica leukotoxin induces expression of inflammatory cytokines from bovine alveolar macrophages. AB - We obtained biologically active purified leukotoxin (Lkt) from Pasteurella haemolytica serotypel, strain 12296 using preparative sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE). Three species of Lkt of molecular masses 95, 100, and 104 kDa were obtained. Purity of all three species of Lkt was confirmed by analytical SDS-PAGE and Western blot (immunoblot) analysis. Results from the chromogenic Limulus amebocyte lysate assay and silver staining of SDS PAGE patterns indicated that the preparations were free of contaminating lipopolysaccharide. We then studied the kinetics of TNF alpha and IL-1 beta mRNA expression in bovine alveolar macrophages stimulated with the purified 104 kDa Lkt. Subcytolytic concentrations of Lkt induced TNF alpha and IL-1 beta gene expression and peak induction was observed at a concentration of 1 leukotoxin unit/ml. Both TNF alpha and IL-1 beta mRNA expression were detectable at 1 h after stimulation with 1 leukotoxin unit/ml. The expression peaked at 2 h, steadily declining up to 6 h, and was undetectable by 10 h. Secreted TNF alpha measured by bioassay peaked at 4-6 h and accumulated at a lesser concentration after 6 h. By contrast, secreted IL-1 peaked at 6 h and decreased significantly by 10 h. The ability of purified Lkt to induce TNF alpha and IL-1 beta gene expression and secretion of bioactive proteins was suppressed by Ca2+ chelating agents, 5 mM EDTA and 5 mM EGTA, but not polymyxin B. Heat-inactivation of the purified Lkt that had lost its cytocidal property completely abrogated induction of TNF alpha and IL-1 beta gene expression and secretion in bovine AMs.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7476091 TI - Mouse neuropathogenic poliovirus strains cause damage in the central nervous system distinct from poliomyelitis. AB - Poliomyelitis as a consequence of poliovirus infection is observed only in primates. Despite a host range restricted to primates, experimental infection of rodents with certain genetically well defined poliovirus strains produces neurological disease. The outcome of infection of mice with mouse-adapted poliovirus strains has been described previously mainly in terms of paralysis and death, and it was generally assumed that these strains produce the same disease syndromes in normal mice and in mice transgenic for the human poliovirus receptor (hPVR-tg mice). We report a comparison of the clinical course and the histopathological features of neurological disease resulting from intracerebral virus inoculation in normal mice with those of murine poliomyelitis in hPVR-tg mice. The consistent pattern of clinical deficits in poliomyelitic transgenic mice contrasted with highly variable neurologic disease that developed in mice infected with different mouse-adapted polioviruses. Histopathological analysis showed a diffuse encephalomyelitis induced by specific poliovirus serotype 2 isolates in normal mice, that affected neuronal cell populations without discrimination, whereas in hPVR-tg animals, damage was restricted to spinal motor neurons. Mouse neurovirulent strains of poliovirus type 2 differed from mouse neurovirulent poliovirus type 1 derivatives in their ability to induce CNS lesions. Our findings indicate that the characteristic clinical appearance and highly specific histopathological features of poliomyelitis are mediated by the hPVR.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7476092 TI - The construction and characterization of colanic acid deficient mutants in an extraintestinal isolate of Escherichia coli (O4/K54/H5). AB - Extraintestinal strains of Escherichia coli possess a variety of virulence factors that enable them to cause disease. These strains express a group 2 capsular polysaccharide which is important in the pathogenic process. Extraintestinal strains evaluated to date are also capable of producing the group 1 capsular polysaccharide colanic acid. The blood isolate CP9 (O4/K54/H5) constitutively produces the group 2, K54 capsule but can be induced to produce colanic acid. In this report we assess whether colanic acid contributes to the pathogenesis of this extraintestinal pathogen. CP9 and its derivatives that are deficient in their ability to produce colanic acid (TR94), the K54 group 2 capsule +/- colanic acid (CP9.137, TR1374) and the O4 specific antigen +/- colanic acid (CP921,CP925) were used to test whether the group 1 capsule colanic acid conferred protection against the bactericidal effects of serum and recombinant bactericidal/permeability-increasing protein (rBPI-23) in vitro. Additionally, CP9, CP9.137 and TR94 were evaluated in the rat granuloma pouch, an in vivo model for localized infection, and by intraperitoneal inoculation into mice, a systemic infection model. In summary, the inability of CP9 to produce colanic acid in the presence or absence of its K54 and O4 antigens had no effect on its ability to survive these host defenses in vitro and did not affect its virulence in these two in vivo models of infection. PMID- 7476093 TI - Experimental Chlamydia pneumoniae infection in mice: effect of reinfection and passive immunization. AB - NIH/S mice were infected intranasally with Chlamydia pneumoniae isolate Kajaani 6 and rechallenged after either 28 or 70 days. A partial resistance to reinfection, indicated by a reduced recovery of live organisms, was noted at both time points of rechallenge: positive isolations from lung homogenates and/or bronchoalveolar lavage fluids were observed in fewer mice and the yields of isolated chlamydiae remained smaller, as compared to primary infection. However, a previous infection did not confer any protection against inflammatory changes. A strong peribronchial and perivascular inflammation with infiltrating lymphocytes and plasma cells was noted in the lungs of primary infected, as well as reinfected, mice. The effect of passive immunization was also studied. When mice were given convalescent or hyperimmune sera intraperitoneally before inoculation, lower C. pneumoniae isolation yields were detected. As in the rechallenge experiment, marked inflammation could still be seen in the lungs, now with polymorphonuclear leukocyte infiltration. The results suggest that immunological reactions play a role in the pathogenesis of C. pneumoniae infection. Antibodies may be important in reducing the amount of infective elementary bodies, but complete clearing of C. pneumoniae could not be achieved in these experiments, even less a protection against inflammatory lung changes. PMID- 7476094 TI - Use of antibiotics to select auxotrophic mutants of Neisseria meningitidis. AB - Antibiotic selection of auxotrophs has been a powerful tool in the elucidation of bacterial metabolic pathways, but it has been difficult to adapt this method to Neisseria spp. We describe a procedure by which a population of mutagenized N. meningitidis is enriched for mutants with specific growth phenotypes. These experiments used a simple defined medium (modified from that described in J Bacteriol 1962; 83: 470-4) in which meningococci grow well on a variety of carbon sources. Nitrosoguanidine-treated meningococci were incubated with an antibiotic (cefotaxime, streptomycin or nalidixic acid) in a defined medium that was nonpermissive for the desired phenotype. The survivors were grown for several generations in a permissive defined medium to reduce the proportion of mutants with phenotypes other than that desired, then subjected to a second antibiotic treatment in nonpermissive medium. Survivors of the second antibiotic treatment were plated, and colonies were screened to identify auxotrophs. This procedure has allowed the isolation of meningococcal mutants with amino acid or vitamin requirements or with altered utilization of carbon sources. PMID- 7476095 TI - Characterization of glycoprotein glycan receptors for Escherichia coli F17 fimbrial lectin. AB - The specificity of N-acetylglucosamine-binding F17 fimbrial lectin of coli (also called FY or Att25) for the oligosacharide structures was investigated by mannose resistant hemagglutination inhibition tests and direct binding assays. The linkage position of N-acetyl-glucosamine influenced its affinity in the preferential order of beta 1-3 > beta 1-6 > beta 1-4 beta > beta 1-2. Minimal GlcNAc beta 1-3Gal beta 1-sequence strongly bound to F17 lectin, whether located in terminal nonreducing position or internally in carbohydrate moieties. F17 lectin specifically interacted with this unit in O-glycosidically linked oligosaccharides of the bovine glycophorins and intestinal mucins. On the contrary, GlcNAc beta 1-NAsn, GlcNAc beta 1-6Man beta- and GlcNAc beta 1-4GlcNAc- of N-glycosylated proteins failed to bind to the lectin. Our findings emphasized the presence of multiple F17 mucosal receptor complex in the newborn calf intestines. Furthermore, the density of receptors for F17 fimbrial lectin seemed to depend on the age of the calf and the intestinal segment. PMID- 7476097 TI - Bacteremia is required for invasion of the murine central nervous system by Listeria monocytogenes. AB - The ability of the facultative intracellular pathogen Listeria monocytogenes to penetrate the central nervous system (CNS) was studied by following the kinetics of brain invasion and histological lesions during an acute intravenous (i.v.) infection in the mouse. CNS invasion occurred during the early phase of infection and produced severe meningoencephalitis characterized by multiple granulomatous foci predominantly located in the brainstem and associated with diffuse meningitis and an intense inflammatory reaction involving the choroid plexuses. Bacterial counts in the brain could reach 10(4.5)-10(5.8) by day 5 of infection with 1-2 x 10(6) bacteria i.v., depending upon the bacterial strain used. It was found that CNS invasion was highly dependent upon the level and the duration of bacteremia, thus indicating that persistent bacteremia is essential to induce meningoencephalitis to L. monocytogenes. PMID- 7476098 TI - An analysis of co-circulating serotypes for bluetongue-17 virulence markers. AB - We have recently identified two markers associated with virulent strains of bluetongue virus serotype 17. These differences are an altered antigenic structure of the outer capsid protein VP2 and an increased electrophoretic mobility of the RNA segment 3 that codes for an inner core protein. We did not observe these markers in confirmed avirulent strains of bluetongue virus serotype 17. We hypothesized that these virulence-associated markers may have been acquired by bluetongue-17 through genetic interaction with other circulating serotypes of the virus. To test this hypothesis, we studied all isolates of other BLU serotypes obtained from the same sentinel cattle herds in Central America and the Caribbean on the same days as BLU-17 isolates. We looked for evidence of common epitopes on VP2 or an RNA segment 3 of identical mobility to that of the virulent strains of BLU-17. We found no evidence to indicate that genetic interaction with other co-circulating serotypes gave rise to these two specific virulence-associated markers of BLU-17. PMID- 7476096 TI - Mutational analysis of the putative leukotoxin transport genes in Actinobacillus actinomycetemcomitans. AB - The periodontal pathogen, Actinobacillus actinomycetemcomitans, produces leukotoxin, a protein that specifically lyses host defense cells. The leukotoxin is similar in sequence and operon organization to the Escherichia coli alpha hemolysin and other members of the RTX family of toxins. However, unlike the other RTX toxins, the A. actinomycetemcomitans leukotoxin is not secreted from the cell and instead remains associated with the outer membrane. Nonetheless, the A. actinomycetemcomitans Ikt operon contains two genes, IktB and IktD, that appear analagous to the toxin localization genes found in the other Gram-negative bacteria. Thus, to determine the roles of these putative transport genes in A. actinomycetemcomitans, we have used insertional mutagenesis to generate mutant strains lacking functional LktB and/or LktD. When either IktD or both IktB and IktD were inactivated, the level of detectable leukotoxin protein in the cell decreased significantly. However, the IktB and IktD mutations had no effect on the levels of leukotoxin RNA. Thus, the lack of LktB and LktD proteins must affect LktA synthesis post-transcriptionally. It is proposed that this is an indirect effect of leukotoxin mislocalization in IktB- and IktD- mutants. Finally, analysis of the mutants revealed that LktB and LktD are not essential for the formation of extracellular membrane vesicles in A. actinomycetemcomitans. PMID- 7476100 TI - Non-dystrophic 129 REJ mice are susceptible to i.p. infection with Listeria monocytogenes despite an ability to recruit inflammatory neutrophils to the peritoneal cavity. AB - In this study we compared the host response to Listeria monocytogenes in 129 REJ mice with listeria-resistant (C57Bl/6j) and susceptible (Balb/c) mouse strains. In all experiments mice were inoculated by the i.p. route. 129 REJ mice and Balb/c mice were sensitive to listeriosis whilst C57Bl/6j mice were relatively resistant to i.p. infection. Relatively large numbers of viable bacteria could be detected in the spleens of 129 REJ mice as early as 6 h following i.p. inoculation suggesting that dissemination of listeria from the peritoneal cavity is rapid in this mouse strain. This contrasted with Balb/c mice which exhibited an early lag phase during which only low numbers of bacteria could be isolated from the spleens of infected animals. In response to both proteose peptone and live listeria, 129 REJ mice demonstrated a greater capacity to recruit neutrophils to the peritoneal cavity than Balb/c and C57Bl/6j mice. In addition, inflammatory phagocytes from 129 REJ mice were as bactericidal in vitro as phagocytes from the Balb/c and C57Bl/6j strains. However, in vivo, inflammatory neutrophils elicited by proteose peptone prior to i.p. infection with L. monocytogenes were not protective in the three mouse strains tested. Despite the apparent inadequacy of peritoneal neutrophils in controlling early bacterial proliferation, depletion of neutrophils in 129 REJ mice severely exacerbated i.p. infection with L. monocytogenes. The results indicate that neutrophils provide an inefficient but essential means of controlling early outgrowth of listeria in the peritoneal cavity of 129 REJ mice. The excessive inflammatory response seen in 129 REJ mice may facilitate the early dissemination of L. monocytogenes from the peritoneal cavity to peripheral sites. PMID- 7476099 TI - Characterisation of group A streptococcal isolates from tropical Australia with high prevalence of rheumatic fever: probing for signature sequences to identify members of the family of serotype 5. AB - The prevalence of rheumatic heart disease (RHD) in Northern Territory Aboriginal communities is high, but there is a low isolation rate of historically rheumatic fever associated M types (such as M5) of group A streptococci (GAS). Many isolates are M non-typable (MNT). Serology suggests that the population is exposed to M5-like isolates; some RHD patients having high IgM or IgG titres to two M5 B-repeat region peptide epitopes, B1 (KQQESK) and B4 (EQKSKQ). To identify relatives of M5 in our collection of GAS, oligonucleotide probes to the B1 and B4 repeat regions shared by M5 and a local M5-like isolate, were used to screen 101 isolates for the presence of signature sequences. In all, 28% of the tropical Australian isolates contained the signature sequences, identifying members of the M5 family. The 5' region of the genes for M proteins from three members of the M5 family fell into two sequence types. Hybridisation to probes based on these sequences suggested that among tropical Australian isolates there are at least three distinct sequence types that contained the M5 signature sequences. These results suggest that a considerable number of M5 family GAS are circulating in tropical Australia. PMID- 7476101 TI - Heterologous production of the P1 porin of Neisseria meningitidis in bacillus subtilis: the effect of an N-terminal extension on the presentation of native like epitopes. AB - The major outer membrane protein P1 (class 1) of Neisseria meningitidis has been produced as inclusion bodies in Bacillus subtilis with the aim to develop a vaccine based on it. The protein produced in high yield in B. subtilis contained an N-terminal extension of 11 amino acid residues which was found to be necessary for expression in the production system. In the present study we asked whether or not the removal of this extension would effect the conformation of this protein in liposomes as judged by its immunogenic properties. A methionine was engineered in front of the mature P1 protein to provide a chemical cleavage site for CNBr to remove the extension. The CNBr-cleaved protein, complexed with phospholipids, elicited high titers of antibodies binding to the meningococcal cells similarly to the noncleaved protein. This suggests that the BacP1 protein can serve as an effective vaccine component irrespective of the presence, or absence, of this N terminal extension. PMID- 7476102 TI - [Parathyroid adenoma and carcinoma. A comparative morphologic study]. AB - A variety of gross, histological and cytological features were analyzed in a comparative study between adenomas and carcinomas of the parathyroid gland. The histological findings more valuable in the differential diagnosis of parathyroid carcinoma were the presence of a thick and irregular capsule infiltrated by the tumor (Fig. 1), broad and irregular intratumoral fibrous septa, and capsular or intratumoral vascular invasion (Table 1). Cytologically, the carcinomas were composed only by chief cells, with a mitotic activity higher than in adenomas, showing the presence of abnormal mitosis (Table 2, Fig 4). Characteristically all carcinomas were composed by a cellular population significantly of greater size than the one observed in adenomas. PMID- 7476103 TI - [Audit of clinical trials in Argentina. Assessment of standardized operational procedures in 12 centers]. AB - The purpose of these audits was to ensure the quality, integrity and validity of investigational data. Audits were performed by control of compliance to standard operational procedures (SOPs) for Good Clinical Practice (GCP). Guidelines were proposed and established by the Food and Drug Administration (FDA) in 1977-1979 and adopted by several countries of the European Community in 1987-1989 and 1990. In Argentina, 12 on site audits of clinical trials performed between 1987 and 1990, were carried out. Basic data of GCP, audited in 58 out of 153 patients included in 12 different centers, were: protection of individual rights, compliance to standard operational procedures, adverse event reports, access to original investigational data and source document archives. The analysis of all audited data showed that patient case reports form recorded findings are credible and reliable and that researchers and monitors had conducted the study with an acceptable level in accordance to SOPs of good clinical practice. Otherwise, several in need of change to obtain better clinical data were identified: ethical committees, written informed consent, protocol adherence, data record, drug accountability, document source archives and the necessity of a thorough theorical and practical training on SOPs for GCP for researchers and monitors of clinical trials. PMID- 7476104 TI - [New findings on Junin virus infection in rodents inside and outside the endemic area of hemorrhagic fever in Argentina]. AB - In conjunction with field trials for a vaccine against Argentine Hemorrhagic Fever (AHF), small mammals were trapped during a 28-month period (1 November 1987 to 13 March 1990) in 3 epidemiologically defined areas of the central Argentine pampas: northern and central Buenos Aires provinces were included in the AHF "historic" area, where the disease was common 15-20 years ago, but case rates are currently low; southern Santa Fe province is the current high-incidence area for AHF; the nonendemic area was represented by two localities 60-90 km beyond the northernmost extension of human disease. Animals were live-trapped for 3 days per month in permanent "mark-recapture" grids in each of the 3 areas. Samples of blood, sera, and oral swabs were taken from these animals before they were marked and released at the site of capture. In addition, "removal" traplines provided animals from 16 localities in these 3 areas which were sacrificed to obtain samples of organs in addition to the aforementioned samples. Samples were tested for the presence of Junin virus (JV) antigen by enzyme immunoassay (ELISA). In this assay, a pool of 13 mouse anti-JV glycoprotein and nucleocapsid monoclonal antibodies adsorbed to the surface of microtiter plates was used to capture JV antigen in sample suspensions. A polyclonal rabbit anti-JV antiserum was added as a detector antibody, and an anti-rabbit antibody conjugated to horseradish peroxidase applied with substrate to complete the sandwich.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7476105 TI - [Diaphragmatic reserve strength in obese patients]. AB - Nine obese patients (OB) and seven normal subjects (N) were studied in order to determine diaphragmatic strength reserve, measured in terms of diaphragm Tension/Time Index (TTdi). This index was measured with the patients awake and during the obstructive apnea (OA) episodes. TTdi was 2.7 times superior in the OB (p < 0.0005) and was related with a lower Pdi Max (102 vs 202 cm H2O; p < 0.005) and with a higher Pdi (9 vs 6 cm H2O; p < 0.05). During OA the TTdi was higher than the threshold value to develop diaphragmatic fatigue (0.15-0.20) in three patients. This value was not exceeded in one patient because of striking paradoxical diaphragmatic movements. As for anthropometric data, DPI% range was higher and wider in OB (Table 1). The pCO2 in OB was 40 +/- 6 mmHg. In all patients (Table 3), severe hypoxia and hypercapnial were observed. TTdi evolution in an OA in 4 patients can be appreciated in Figure 3. It can be concluded that the diaphragmatic strength reserve is reduced in OB so that they are more susceptible to develop diaphragmatic fatigue. A fatigant respiratory pattern was also registered during the OA. PMID- 7476106 TI - Morphometry of skeletal muscle involvement in mice infected or preimmunized with live attenuated Trypanosoma cruzi. AB - A morphometric study was undertaken in the quadriceps muscle of Swiss mice in order to assess the effects of immunization with attenuated T. cruzi upon tissue lesions. interfascicular lymphocytic infiltration, presence of amastigote nests, vascular lesions, degeneration and fibrosis were evaluated independently. Each of these alterations was drastically prevented in preimmunized animals. These results indicate that immunity against T. cruzi not only reduces circulating parasites but also clears most of the organic damage caused by infection. PMID- 7476107 TI - [Renal angiomyolipoma]. AB - Five cases of angiomyolipoma with acute renal symptomatology have been studied using different diagnostic methods such as intravenous urogram, ultrasound, computed axial tomography. These procedures permit an adequate diagnosis of angiomyolipoma. This benign tumor may produce important symptomatology when bleeding reaches into the perinephral as well as retroperitoneal space. The identification of fat tissue within the renal tumoral lesion confirms the diagnosis of angiomyolipoma. PMID- 7476109 TI - [HIV infection in a patient with chronic cutaneous lupus]. AB - We present a case of chronic cutaneous lupus erythematosus (CCLE) in a patient who subsequently became infected with human immunodeficiency virus, HIV-1. The symptoms attributed to CCLE had persisted for years, and curiously had disappeared by the time the patient probably became infected with HIV-1. Even if there is lack of evidence to consider HIV-1 infection as direct causative agent in the clinical remission of CCLE, this response is in accordance with previous reports from other authors, regarding the attenuating role of HIV on the clinical course of the systemic lupus erythematosus. Even if the number of reported cases is very low it is of the upmost importance to establish whether this hypothesis is correct or not, since both diseases present great difficulties regarding diagnosis, due to the important overlapping between symptoms and serological tests. PMID- 7476108 TI - [Effects of cold on triiodothyronine metabolism in the pregnant rat]. AB - Sixteen Wistar rats were studied in the last week of pregnancy. Another group of 6 nonpregnant animals served as control. Eight pregnant and 6 control ratas were kept in the cold room at 4 degrees C for 4 days, while 8 pregnant rats remained at 22 degrees C. After blocking the thyroid gland with potassium perchlorate to prevent iodine reutilization, a dose of 4 microCi of 125I-T3 was injected i.p. Thereafter, animals were placed in individual metabolic cages for separate 24 h collection of urine and feces. Heparinized blood samples were obtained at 4, 14, 24 and 28 h following tracer injection. Pregnant rats exposed to cold had a significant increase in T3 fractional turnover as compared to pregnants at 22 degrees C (p < 0.005) and to controls at 4 degrees C (p < 0.02). No changes were observed in space of distribution, serum concentration and body pool of T3. Degradation of this hormone, however, was significantly augmented (p < 0.05) in pregnants at 4 degrees C as compared to pregnants at 22 degrees C, although not when compared to controls at 4 degrees C. The urinary excretion of 125I was increased in pregnants in the cold (p <0.005) versus pregnants at 22 degrees C. The metabolic (p < 0.005), urinary (p < 0.001) and fecal (p < 0.05) clearances in pregnants at 4 degrees C were augmented with respect to the other two groups.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7476111 TI - [Wegener's granulomatosis with a 20-year course]. PMID- 7476110 TI - [Spinal myoclonus secondary to the intravenous administration of iodine contrast media]. AB - We describe a patient with a longstanding paraplegia who developed spinal myoclonus on 3 different occasions spanning one year, once after an enhanced CT scan and twice after excretory urographies, one of which was also followed by a generalized tonic-clonic seizure. To our knowledge only one case of spinal myoclonus secondary to the administration of intravenous contrast material in a patient with a spinal arteriovenous malformation has yet been reported. Taken together, the findings in these cases suggest that spinal myoclonus following intravenous iodine administration is indicative of an underlying spinal cord lesion. PMID- 7476112 TI - [A new levodopa benserazide preparation for Parkinson's disease with motor fluctuations refractory to standard L-dopa]. AB - As Parkinson's disease worsens, many patients develop motor fluctuations which usually correlate directly or indirectly with L-dopa plasma levels. A new L-dopa benserazide HBS preparation (Madopar) a control release pharmaceutical formulation which is activated when it contacts gastric fluid thus providing more stable L-dopa plasma levels, was assayed. Ten patients with a diagnosis of idiopathic Parkinson's disease and motor fluctuations otherwise unresponsive to conventional therapy were selected. The average age was 62 years and the duration of the disease 9 years. The motor status was evaluated on an hourly basis with the King's College Parkinson's disease rating scale; in addition, a nocturnal disability scale (Lees) was used. Out of the 10 patients, 2 dropped out within the first month due to worsening of parkinsonian signs, while 7 of the remainders preferred HBS preparation to the previous treatment. The number of off hours in this group was reduced by 58% and motor fluctuation became less severe. In only 3 cases was it possible to use HBS as monotherapy while in the rest standard L-dopa had to be added, specially as morning doses. The average L-dopa daily dose was increased by 36%. Unwanted effects included psychiatric disturbances in two (in one L-dopa dose had to be reduced) and epigastralgia in one. Our findings suggest that this L-dopa-benserazide control release may be considered an able therapeutic formulation in the control of motor fluctuations in Parkinson's disease. PMID- 7476113 TI - [The structured objective clinical examination. An instrument for the evaluation of competence]. PMID- 7476114 TI - [The 1991 Nobel Price in Medicine; the ionic channels of biologic membranes]. PMID- 7476116 TI - [The fixed period]. PMID- 7476115 TI - [Good clinical practices]. PMID- 7476117 TI - [Cat scratch disease: 2 cases with argyrophil bacilli]. PMID- 7476118 TI - [Convulsions induced ty OKT3?]. PMID- 7476119 TI - Motoo Kimura (1924-1994). PMID- 7476120 TI - Spread of the autonomous transposable element hobo in the genome of Drosophila melanogaster. AB - The transposable element hobo has been introduced into the previously empty Drosophila melanogaster strain Hikone so that its dynamics can be followed and it can be compared with the P element. Five transformed lines were followed over 58 generations. The results were highly dependent on the culture temperature, the spread of hobo element being more efficient at 25 degrees C. The multiplication of hobo sequences resulted in a change in the features of these lines in the hobo system of hybrid dysgenesis. The number of hobo elements remained low (two to seven copies) and the insertions always corresponded to complete sequences. Our findings suggest that, despite their genetic similarities, P and hobo elements differ in many aspects, such as mobility and regulation mechanisms. PMID- 7476121 TI - Male and female mitochondrial DNA lineages in the blue mussel (Mytilus edulis) species group. AB - In blue mussels of the Mytilus edulis species complex, mitochondrial DNA (mtDNA) inheritance is coupled with gender. Females receive their mother's mtDNA and pass it on to both their daughters and sons. In addition, males receive mtDNA from their father and transmit this male mtDNA to their sons. If this pattern of "doubly uniparental inheritance" is older than the M. edulis species complex, then all members of this group must have two distinct mtDNA lineages: a maternal lineage that is transmitted through females and a paternal lineage that is transmitted through males. To test this hypothesis, we scored mtDNA variation in two taxa in this complex, M. edulis and M. trossulus, by means of restriction fragment profiles of whole-mtDNA genomes and DNA sequence of a region of the cytochrome c oxidase subunit III gene (COIII). The various mitotypes present in these mussels were classified as "male" or "female" based on their gender association and as belonging to M. edulis or M. trossulus based on species specific allozymes. Both maximum parsimony and neighbor-joining phylogenies based on the COIII sequences grouped female and male mtDNAs into two distinct lineages irrespective of specific origin in accordance with the hypothesis that the origin of these lineages predates the divergence of M. edulis and M. trossulus. PMID- 7476122 TI - Studies on the transposition rates of mobile genetic elements in a natural population of Drosophila melanogaster. AB - In an isolated population of Drosophila melanogaster on Ishigaki Island the chromosomal distribution of several retrotransposons, including copia, 412, 297, 17.6, I, and jockey elements, was examined by in situ hybridization. In this population the cosmopolitan inversion, In(2L)t, is known to exist in high frequency. One major haplotype concerning the occupied sites of the transposable elements was identified in the In(2L)t-carrying chromosomes. This haplotype is suggested to be the ancestral one. The age of the inversion in this local population was estimated to be 1,400 generations. The transposition rates of these elements were estimated based on the age of the inversion and the number of the elements lost and gained. The excision rates were in the range from 9.13 x 10(-5) to 2.25 x 10(-4) per site per generation. They were similar each other in the copia-like elements as well as in the LINE-like elements. The rate was higher in the copia-like elements than in the LINE-like elements. Insertions occurred in the range from 6.79 x 10(-4) to 9.05 x 10(-4) per element per generation. It is herein shown that both insertions and excisions occurred at a significantly higher rate in this population than in the laboratory. PMID- 7476123 TI - Variations in mitochondrial tRNA gene organization of reptiles as phylogenetic markers. AB - Amplification and sequencing of mitochondrial DNA regions corresponding to three major clusters of transfer RNA genes from a variety of species representing major groups of birds and reptiles revealed some new variations in tRNA gene organization. First, a gene rearrangement from tRNA(His)-tRNA(Ser)(AGY) tRNA(Leu)(CUN) to tRNA(Ser)(AGY)-tRNA(His)tRNA(Leu)(CUN) occurs in all three crocodilians examined (alligator, caiman, and crocodile). In addition an exceptionally long spacer region between the genes for NADH dehydrogenase subunit 4 and tRNA(Ser)(AGY) is found in caiman. Second, in congruence with a recent finding by Seutin et al., a characteristic stem-and-loop structure for the putative light-strand replication origin located between tRNA(Asn) and tRNA(Cys) genes is absent for all the birds and crocodilians. This stem-and-loop structure is absent in an additional species, the Texas blind snake, whereas the stem-and loop structure is present in other snakes, lizards, turtles, mammals, and a frog. The disappearance of the stem-and-loop structure in the blind snake most likely occurred independently of that on the lineage leading to birds and crocodilians. Finally, the blind snake has a novel type of tRNA gene arrangement in which the tRNA(Gln) gene moved from one tRNA cluster to another. Sequence substitution rates for the tRNA genes appeared to be somewhat higher in crocodialians than in birds and mammals. As regards the controversial phylogenetic relationship among the Aves, Crocodilia, and Mammalia, a sister group relationship of birds and crocodilians relative to mammals, as suggested from the common loss of the stem and-loop structure, was supported with statistical significance by molecular phylogenetic analyses using the tRNA gene sequence data. PMID- 7476124 TI - Evidence for independent recruitment of zeta-crystallin/quinone reductase (CRYZ) as a crystallin in camelids and hystricomorph rodents. AB - Zeta-crystallin/quinone reductase (CRYZ) is an NADPH oxidoreductase expressed at very high levels in the lenses of two groups of mammals: camelids and some hystricomorph rodents. It is also expressed at very low levels in all other species tested. Comparative analysis of the mechanisms mediating the high expression of this enzyme/crystallin in the lens of the Ilama (Lama guanacoe) and the guinea pig (Cavia porcellus) provided evidence for independent recruitment of this enzyme as a lens crystallin in both species and allowed us to elucidate for the first time the mechanism of lens recruitment of an enzyme-crystallin. The data presented here show that in both species such recruitment most likely occurred through the generation of new lens promoters from nonfunctional intron sequences by the accumulation of point mutations and/or small deletions and insertions. These results further support the idea that recruitment of CRYZ resulted from an adaptive process in which the high expression of CRYZ in the lens provides some selective advantage rather than from a purely neutral evolutionary process. PMID- 7476125 TI - Phylogenetic place of mitochondrion-lacking protozoan, Giardia lamblia, inferred from amino acid sequences of elongation factor 2. AB - Partial regions of the mRNA encoding a major part of translation elongation factor 2 (EF-2) from a mitochondrion-lacking protozoan, Giardia lamblia, were amplified by polymerase chain reaction, and their primary structures were analyzed. The deduced amino acid sequence was aligned with other eukaryotic and archaebacterial EF-2's, and the phylogenetic relationships among eukaryotes were inferred by the maximum likelihood (ML) and the maximum parsimony (MP) methods. The ML analyses using six different models of amino acid substitutions and the MP analysis consistently suggest that among eukaryotic species being analyzed, G. lamblia is likely to have diverged from other higher eukaryotes on the early phase of eukaryotic evolution. PMID- 7476126 TI - Natural selection on the gag, pol, and env genes of human immunodeficiency virus 1 (HIV-1). AB - Natural selection on polymorphic protein-coding loci of human immunodeficiency virus-1 (HIV-1), the more geographically widespread of the two viruses causing human acquired immune deficiency syndrome (AIDS), was studied by estimating the rates of nucleotide substitution per site in comparisons among alleles classified in families of related alleles on the basis of a phylogenetic analysis. In the case of gag, pol, and gp41, the rate of synonymous substitution generally exceeded that of nonsynonymous substitution, indicating that these genes are subject to purifying selection. However, in the case of several of the variable (V) regions of the gp120 gene, especially V2 and V3, comparisons within and between families often showed a significantly higher rate of nonsynonymous than of synonymous nucleotide substitution. This pattern of nucleotide substitution indicates that positive Darwinian selection has acted to diversify these regions at the amino acid level. The V regions have been identified as probable epitopes for antibody recognition; therefore, avoidance of such recognition seems likely to be the basis for positive selection on these regions. By contrast, regions of HIV-1 proteins identified as epitopes for T cell recognition show no evidence of positive selection and are often highly conserved at the amino acid level. These results suggest that selection favoring avoidance of T cell recognition has not been a major factor in the history of HIV-1 and thus that avoidance of T cell recognition is not likely to be a major factor in the pathogenesis of AIDS. PMID- 7476127 TI - Sampling properties of DNA sequence data in phylogenetic analysis. AB - We inferred phylogenetic trees from individual genes and random samples of nucleotides from the mitochondrial genomes of 10 vertebrates and compared the results to those obtained by analyzing the whole genomes. Individual genes are poor samples in that they infrequently lead to the whole-genome tree. A large number of nucleotide sites is needed to exactly determine the whole-genome tree. A relatively small number of sites, however, often results in a tree close to the whole-genome tree. We found that blocks of contiguous sites were less likely to lead to the whole-genome tree than samples composed of sites drawn individually from throughout the genome. Samples of contiguous sites are not representative of the entire genome, a condition that violates a basic assumption of the bootstrap method as it is applied in phylogenetic studies. PMID- 7476128 TI - Phylogenetic test of the molecular clock and linearized trees. AB - To estimate approximate divergence times of species or species groups with molecular data, we have developed a method of constructing a linearized tree under the assumption of a molecular clock. We present two tests of the molecular clock for a given topology: two-cluster test and branch-length test. The two cluster test examines the hypothesis of the molecular clock for the two lineages created by an interior node of the tree, whereas the branch-length test examines the deviation of the branch length between the tree root and a tip from the average length. Sequences evolving excessively fast or slow at a high significance level may be eliminated. A linearized tree will then be constructed for a given topology for the remaining sequences under the assumption of rate constancy. We have used these methods to analyze hominoid mitochondrial DNA and drosophilid Adh gene sequences. PMID- 7476129 TI - Body temperature, rate of biosynthesis, and evolution of genome size. AB - An optimality model relating the rate of biosynthesis to body temperature and gene duplication is presented to account for several observed patterns of genome size variation. The model predicts (1) that poikilotherms living in a warm climate should have a smaller genome than poikilotherms living in a cold climate, (2) that homeotherms should have a small genome as well as a small variation in genome size relative to their poikilothermic ancestors, (3) that cold geological periods should favor the evolution of poikilotherms with a large genome and that warm geological periods should do the opposite, and (4) that poikilotherms with a small genome should be more sensitive to changes in temperature than poikilotherms with a large genome. The model also offers two explanations for the empirically documented trend that organisms with a large cell volume have larger genomes than those with a small cell volume. Relevant empirical evidence is summarized to support these predictions. PMID- 7476130 TI - The robustness of two phylogenetic methods: four-taxon simulations reveal a slight superiority of maximum likelihood over neighbor joining. AB - The robustness (sensitivity to violation of assumptions) of the maximum likelihood and neighbor-joining methods was examined using simulation. Maximum likelihood and neighbor joining were implemented with Jukes-Cantor, Kimura, and gamma models of DNA substitution. Simulations were performed in which the assumptions of the methods were violated to varying degrees on three model four taxon trees. The performance of the methods was evaluated with respect to ability to correctly estimate the unrooted four-taxon tree. Maximum likelihood outperformed neighbor joining in 29 of the 36 cases in which the assumptions of both methods were satisfied. In 133 of 180 of the simulations in which the assumptions of the maximum-likelihood and neighbor-joining methods were violated, maximum likelihood outperformed neighbor joining. These results are consistent with a general superiority of maximum likelihood over neighbor joining under comparable conditions. They extend and clarify an earlier study that found an advantage for neighbor joining over maximum likelihood for gamma-distributed mutation rates. PMID- 7476132 TI - Improved analyses of human mtDNA sequences support a recent African origin for Homo sapiens. AB - New quantitative methods are applied to the 135 human mitochondrial sequences from the Vigilant et al. data set. General problems in analyzing large numbers of short sequences are discussed, and an improved strategy is suggested. A key feature is to focus not on individual trees but on the general "landscape" of trees. Over 1,000 searches were made from random starting trees with only one tree (a local optimum) being retained each time, thereby ensuring optima were found independently. A new tree comparison metric was developed that is unaffected by rearrangements of trees around many very short internal edges. Use of this metric showed that downweighting hypervariable sites revealed more evolutionary structure than studies that weighted all sites equally. Our results are consistent with convergence toward a global optimum. Crucial features are that the best optima show very strong regional differentiation, a common group of 49 African sequences is found in all the best optima, and the best optima contain the 16 !Kung sequences in a separate group of San people. The other 86 sequences form a heterogeneous mixture of Africans, Europeans, Australopapuans, and Asians. Thus all major human lineages occur in Africa, but only a subset occurs in the rest of the world. The existence of these African-only groups strongly contradicts multiregional theories for the origin of Homo sapiens that require widespread migration and interbreeding over the entire range of H. erectus. Only when the multiregional model is rejected is it appropriate to consider the root, based on a single locus, to be the center of origin of a population (otherwise different loci could give alternative geographic positions for the root). For this data, several methods locate the root within the group of 49 African sequences and are thus consistent with the recent African origin of H. sapiens. We demonstrate that the time of the last common ancestor cannot be the time of major expansion in human numbers, and our results are thus also consistent with recent models that differentiate between the last common ancestor, expansion out of Africa, and the major expansion in human populations. Such a two-phase model is consistent with a wide range of molecular and archeological evidence. PMID- 7476131 TI - Recent horizontal transfer of a mariner transposable element among and between Diptera and Neuroptera. AB - Transposable elements of the mariner family are widespread among insects and other invertebrates, and initial analyses of their relationships indicated frequent occurrence of horizontal transfers between hosts. A specific PCR assay was used to screen for additional members of the irritans subfamily of mariners in more than 400 arthropod species. Phylogenetic analysis of cloned PCR fragments indicated that relatively recent horizontal transfers had occurred into the lineages of a fruit fly Drosophila ananassae, the horn fly Haematobia irritans, the African malaria vector mosquito Anopheles gambiae, and a green lacewing Chrysoperla plorabunda. Genomic dot-blot analysis revealed that the copy number in these species varies widely, from about 17,000 copies in the horn fly to three copies in D. ananassae. Multiple copies were sequenced from genomic clones from each of these species and four others with related elements. These sequences confirmed the PCR results, revealing extremely similar elements in each of these four species (greater than 88% DNA and 95% amino acid identity). In particular, the consensus sequence of the transposase gene of the horn fly elements differs by just two base pairs out of 1,044 from that of the lacewing elements. The mosquito lineage has diverged from the other Diptera for over 200 Myr, and the neuropteran last shared a common ancestor with them more than 265 Myr ago, so this high similarity implies that these transposons recently transferred horizontally into each lineage. Their presence in only the closest relatives in at least the lacewing lineage supports this hypothesis. Such horizontal transfers provide an explanation for the evolutionary persistence and widespread distribution of mariner transposons. We propose that the ability to transfer horizontally to new hosts before extinction by mutation in the current host constitutes the primary selective constraint maintaining the sequence conservation of mariners and perhaps other DNA-mediated elements. PMID- 7476133 TI - A common group I intron between a plant parasitic fungus and its host. AB - The self-splicing RNAs known as group I introns exist in many organisms, but their distribution is difficult to explain. We hypothesize that group I introns have been transferred between a parasite and its host. We describe here the discovery of a common group I intron sequence between a plant-parasitic fungus, Protomyces inouyei, and its host, Youngia japonica. It strongly supports our theory that the group I intron had been transferred from the host plant to the parasitic fungus in the course of evolution. PMID- 7476134 TI - The number of segregating sites in expanding human populations, with implications for estimates of demographic parameters. AB - The frequency distribution of pairwise differences between sequences of mtDNA has recently been used to estimate the size of human populations before and after a hypothetical episode of rapid population growth and the time at which the population grew. To test the internal consistency of this method, we used three different sets of human mtDNA data and the corresponding demographic parameters estimated from the distribution of pairwise differences to determine by simulation the expected number of segregating sites, S, and its empirical distribution. The results indicate that the observed values of S are significantly lower than expected in two of three cases under the assumption of the infinite-sites model. Further simulations in which mutations were allowed to occur more than once at the same site and in which there was variation in mutation rate among sites show that the expected number of segregating sites can be much lower than under the infinite-site assumption. Nevertheless, the observed value of S is still significantly different from the value expected under the expansion hypothesis in two of three cases. PMID- 7476135 TI - Evolutionary relationships among the male and female mitochondrial DNA lineages in the Mytilus edulis species complex. AB - A novel form of mitochondrial DNA (mtDNA) inheritance has previously been documented for the blue mussel (Mytilus edulis). Female mussels inherit their mtDNA solely from their mother while males inherit mtDNA from both their mother and their father. In males, the paternal mtDNA is preferentially amplified so that the male gonad is highly enriched for the paternal mtDNA that is then transmitted from fathers to sons. We demonstrate that this mode of mtDNA inheritance also operates in the closely related species M. galloprovincialis and M. trossulus. The evolutionary relationship between the male and female mtDNA lineages is estimated by phylogenetic analysis of 455 nucleotides from the large subunit ribosomal RNA gene. We have found that the male and female lineages are highly divergent; the divergence of these lineages began prior to the speciation of the three species of blue mussels. Further, the separation between the male and female lineages is estimated to have occurred between 5.3 and 5.7 MYA. PMID- 7476136 TI - Molecular evolution of P transposable elements in the genus Drosophila. I. The saltans and willistoni species groups. AB - A phylogenetic survey using the polymerase chain reaction (PCR) has identified four major P element subfamilies in the saltans and willistoni species groups of Drosophila. One subfamily, containing about half of the sequences studied, consists of elements that are very similar to the canonical (and active) P element from D. melanogaster. Within this subfamily, nucleotide sequence differentiation among different copies from the same species and among elements from different species is relatively low. This observation suggests that the canonical elements are relatively recent additions to the genome or, less likely, are evolving slowly relative to the other subfamilies. Elements belonging to the three noncanonical lineages are distinct from the canonical elements and from one another. Furthermore, there is considerably more sequence variation, on the average, within the noncanonical subfamilies compared to the canonical elements. Horizontal transfer and the coexistence of multiple, independently evolving element subfamilies in the same genome may explain the distribution of P elements in the saltans and willistoni species groups. Such explanations are not mutually exclusive, and each may be involved to varying degrees in the maintenance of P elements in natural populations of Drosophila. PMID- 7476137 TI - A novel measure of genetic distance for highly polymorphic tandem repeat loci. AB - Genetic distance measures are indicators of relatedness among populations or species and are useful for reconstructing the historic and phylogenetic relationships among such groups. Classical measures of genetic distance were developed to analyze biochemical and serological polymorphisms, systems which generally show limited variability. However, these traditional measures of genetic distance are inadequate for the analysis of certain classes of variable number tandem repeat (VNTR) loci, which have a larger number of alleles and higher levels of heterozygosity than traditional genetic markers. At the higher levels of heterozygosity observed at these loci, the standard measures of genetic distance are nonlinear and do not account for the mutational mechanisms of hypervariable loci. We have developed a measure of genetic distance, DSW, which is appropriate for the analysis of highly polymorphic DNA loci. Using computer simulations of diverging populations, we show that DSW conforms to linearity and that the variance is similar in magnitude to traditional measures of genetic distance. Comparisons of phylogenetic trees derived from the simulated divergence of human racial groups demonstrate that the branch lengths of trees prepared using DSW are more similar to the model tree than those generated using other measures. Finally, we demonstrate the applicability of DSW to evolutionary analysis by reconstructing the relationships among eight human populations using 14 microsatellite and STR loci. The phylogenetic trees generated using DSW are different from trees constructed with traditional measures and better reflect the well-documented ancient divergence of African and non-African populations. PMID- 7476138 TI - Maximum-likelihood estimation of molecular haplotype frequencies in a diploid population. AB - Molecular techniques allow the survey of a large number of linked polymorphic loci in random samples from diploid populations. However, the gametic phase of haplotypes is usually unknown when diploid individuals are heterozygous at more than one locus. To overcome this difficulty, we implement an expectation maximization (EM) algorithm leading to maximum-likelihood estimates of molecular haplotype frequencies under the assumption of Hardy-Weinberg proportions. The performance of the algorithm is evaluated for simulated data representing both DNA sequences and highly polymorphic loci with different levels of recombination. As expected, the EM algorithm is found to perform best for large samples, regardless of recombination rates among loci. To ensure finding the global maximum likelihood estimate, the EM algorithm should be started from several initial conditions. The present approach appears to be useful for the analysis of nuclear DNA sequences or highly variable loci. Although the algorithm, in principle, can accommodate an arbitrary number of loci, there are practical limitations because the computing time grows exponentially with the number of polymorphic loci. Although the algorithm, in principle, can accommodate an arbitrary number of loci, there are practical limitations because the computing time grows exponentially with the number of polymorphic loci. PMID- 7476139 TI - Phylogenetic relationships of amphibian families inferred from DNA sequences of mitochondrial 12S and 16S ribosomal RNA genes. AB - Nucleotide sequence comparisons were used to investigate ordinal and familial relationships within the class Amphibia. Approximately 850 base pairs of the mitochondrial 16S ribosomal RNA (rRNA) gene from representatives of 28 of the 40 families of extant amphibians were sequenced. Phylogenetic analyses of these data together with published data of the 12S rRNA gene for the same families and both genes for three more taxa (approximately 1,300 base pairs total for 35 taxa) support the monophyly of each of the three amphibian orders: Anura (confidence value with the interior-branch test: P(c) = 99%), Caudata (P(c) = 100%), and Gymnophiona (P(c) = 99%). An analysis using the four-cluster method cannot discriminate significantly between all three possible unrooted trees involving the three orders of amphibians and an outgroup. Within the Anura, there is support for the monophyly of the two suborders: Neobatrachia (P(c) = 100%) and Archaeobatrachia (P(c) = 97%); the latter was believed to be paraphyletic on the basis of morphology. Within the Archaeobatrachia, the following pairs of taxa cluster: Pelobatidae + Pelodytidae (P(c) = 99%), Pipidae + Rhinophrynidae (P(c) = 99%), Ascaphus + Leiopelmatidae (P(c) = 89%), and Bombina + Discoglossidae (P(c) = 99%). The latter six taxa cluster (P(c) = 94%) such that Pelobatidae + Pelodytidae forms a basal lineage within the Archaeobatrachia. Three major lineages are distinguished within the Neobatrachia: the superfamily Bufonoidea sensu Duellman (P(c) = 86%), the superfamily Ranoidea sensu Lynch (P(c) = 99%), and the Sooglossidae. Basal within the Bufonoidea, Myobatrachidae + Heleophrynidae cluster at P(c) = 96%. The enigmatic Dendrobatidae clusters with the bufonoid families (P(c) = 92%) and is excluded from the ranoid families (P(c) = 99%). (ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7476140 TI - Interchromosomal exchange of genetic information between gene arrangements on the third chromosome of Drosophila pseudoobscura. AB - During the last 60 years, the inversion polymorphism on the third chromosome of Drosophila pseudoobscura has become a case study of the evolution of linked blocks of genes, isolated from each other by the suppression of recombination in heterozygotes for different inversions. Due to its location within inverted regions in most gene arrangements, the amylase (Amy) gene region can be used to elucidate the molecular pattern of evolution in these inversions. We studied this region in the Tree Line phylad of gene arrangements, with regard to both restriction site polymorphisms (RSP) and nucleotide sequences. The analysis of restriction maps, encompassing 26 kb, corroborates the cytogenetic phylogeny established on the basis of inversion breakpoints. However, we found that the 2.7 kb of nucleotide sequences of the AmyI gene are identical in both Estes Park and Hidalgo arrangements, despite the fact that these inversions arose independently from Tree Line. These contrasting results suggest that a homogenizing force, most likely gene conversion, is able to bring about localized exchanges between otherwise isolated gene arrangements. PMID- 7476141 TI - Nucleotide sequence of a marsupial LINE-1 element and the evolution of placental mammals. AB - L1 (LINE-1) elements, previously described in several placental species, were shown to exist in several thousands of copies dispersed in the genome of a marsupial (the North American opossum, Didelphis virginiana). A restriction fragment containing the partial nucleotide sequence of the reverse transcriptase gene (ORF-2) from the marsupial L1 element was cloned and sequenced. The element was shown to evolve in concert within the marsupial when compared to placental mammals. When the marsupial L1 element is used as an outgroup to root a phylogeny of placental mammals, the results support the view that humans more recently shared a common ancestor with ungulates than with rodents. PMID- 7476142 TI - A stem-loop "kissing" model for the initiation of recombination and the origin of introns. AB - Mutations which improve the efficiency of recombination should affect either the proteins which mediate recombination or their substrate, DNA itself. The former mutations would be localized to a few sites. The latter would be dispersed. Studies of hybridization between RNA molecules have suggested that recombination may be initiated by a homology search involving the "kissing" of the tips of stem loops. This predicts that, in the absence of other constraints, mutations which assist the formation of stem loops would be favored. From comparisons of the folding of normal and shuffled DNA sequences, I present evidence for an evolutionary selection pressure to distribute stem loops generally throughout genomes. I propose that this early pressure came into conflict with later local pressures to impose information concerning specific function. The conflict was accommodated by permitting sections of DNA concerned with a specific function to evolve in dispersed segments. Traces of the conflict seem to be present in some modern intron-containing genes. Thus, introns may have allowed the interspersing of selectively advantageous stem loops in coding regions of DNA. PMID- 7476143 TI - An improved method for estimating the rate of fixation of favorable mutations based on DNA polymorphism data. PMID- 7476144 TI - Evidence against concerted evolution in a tandem array in the clumping factor gene of Staphylococcus aureus. PMID- 7476145 TI - [Mortality among workers of the rubber industry. III. Results of further observation of the male cohort]. AB - Mortality among workers of the rubber industry was assessed following the observation of the cohort comprised of 6,978 male workers who had started their employment in the plant producing rubber footwear during the years 1945-1973, and worked for, at least, three months. The condition of the cohort was assessed for December 31, 1990. Standardised mortality rate (SMR) was used as a measurement tool and it was calculated by means of the man-year method. The general population of Poland was taken as the reference population. General mortality in the cohort was significantly higher than in the reference population (2020 death, SMR = 110). Significant excess mortality due to atherosclerosis (205 deaths, SMR = 135) and cirrhosis of the liver (48 deaths, SMR = 170) was also noted. Total number of deaths due to malignant neoplasms-421-was slightly higher than expected. Significant excess of the bladder cancer (13 deaths, SMR = 357), the larynx cancer (23 deaths, SMR = 180) and the lung cancer (148 deaths, SMR = 122) was revealed. Significantly increased risk of the large intestine cancer (15 deaths, SMR = 242) was observed in the subcohort of workers employed in direct production departments. PMID- 7476146 TI - [Exposure to mineral fibers in selected ceramics plants]. AB - The aim of the work was to check, in the selected ceramic plants, whether the replacement of asbestos heat-insulating material by mixed material or fibrous material free from asbestos decreased the risk of exposure to mineral fibre dust among workers engaged in the overhaul of furnace trucks and ceramic furnaces. In the 6 selected plants which used various heat-insulating materials, dust concentration was measured (personal dosimetry), composition of mineral dust, collected from air at overhaul posts, was identified (RTG and IR methods), and morphology of dust particles was examined (optic microscopy and TEM). The highest concentration of respirable mineral fibres was found during the overhaul of a truck with asbestos insulation; respirable fibre concentration reached the value of 5 f/cm3, and total dust concentration-the value of about 80 mg/m3. Insulating plates made of asbestos paper-board proved to be the major source of the mineral fibre emission; mean weighted dust concentration in the plants under study ranged from 2.0 to 2.7 f/cm3. The replacement of asbestos plates by plates made of other heat-insulating materials have lowered respirable mineral fibre dust concentration by ten times. In dust samples collected from the air at the overhaul workposts, the presence of chrysotile, forsterite, quartz and cristobalite (asbestos heat-insulating materials) as well as cristobalite and mullite (non-asbestos heat-insulating materials) was found. PMID- 7476147 TI - [Concentration of dihydroepiandrosterone, dihydroepiandrosterone sulphate and testosterone during premature menopause in women chronically exposed to carbon disulfide]. AB - The authors examined clinically 1710 women. They were split into two groups depending on the type of work performed. Group I (mean age = 37.3 years; average employment period = 26.2 years) was not exposed to carbon disulfide. The incidence of premature menopause in this group was 8.05%. Group II (n = 717; mean age = 38.3 years; average employment period = 25.6 years) was exposed to CS2 concentration in the air ranging from 9.36 to 23.4 mg/m3. In this group 119 women (16.5%) showed symptoms of premature menopause. Concentration of androgens in serum was determined using radioimmunoassay. It was found that women chronically exposed to carbon disulfide had significantly lowered values for dihydroepiandrosterone sulphate (p < 0.001), total testosterone (p < 0.01) and free testosterone (p < 0.01) when compared with the controls. No difference was found in the concentration of dihydroepiandrosterone. PMID- 7476148 TI - [Functional disturbances of the nervous system in the practice of occupational health]. AB - On the basis of a questionnaire survey performed in the group of 80 persons, the author defined how the Occupational Health Service physicians conceive functional disturbances of the nervous system in their daily practice. The survey shows that they are not trained enough to be able to assess this kind of sickness. Their work is hindered by a lack of generally adopted definitions, classification of diseases and methodological guidelines how to define working ability of persons with functional disturbances of the nervous system. PMID- 7476149 TI - [Methods for determining lead and cadmium in blood; cadmium, copper, nickel and chromium in urine using flameless atomic absorption spectrometry]. AB - The estimation of environmental and occupational exposure to metals is based on the determination of metal concentrations in biological material. This paper describes methods for determining lead and cadmium in blood, and cadmium, copper, nickel and chromium in urine. The methods are evaluated in view of their linearity, precision, repeatability, reproducibility and detectability. The reliability of the methods was verified by determining lyphilized standards of known metal concentrations (Behring). The reproducibility expressed by the value of relative standard deviation ranged from 1.2% to 7.8%. In the case of lead and cadmium determinations, the reliability of results was confirmed due to the participation of our laboratory in the UK NEQAS external control project, the Queen Elizabeth Hospital, Birmingham. The results of external quality control (MR VIS = 19 for lead, as compared with the mean = 41 for all the project participants and MR VIS = 29 for cadmium, mean = 62) prove undoubtedly correctness of the methods applied. A small volume of sample to be analyzed is their major advantage, and due to adopted analytical parameters the methods can be used to determine metals in biological material in order to assess occupational or environmental exposure. PMID- 7476150 TI - [Alkyl derivatives of benzene, indene, naphthalene, diphenyl and fluorene as a potential source of occupational and environmental exposure]. AB - Mixtures of organic solvents are used extensively in industry. The aim of this study was to develop a method for the determination of components of commercial multi-component solvent mixtures, and to perform qualitative and quantitative analyses of the mixtures. Farbasol, Solvesso and Shellsol mixtures were analysed by gas chromatography with a mass spectrometry detector (GC/MS). The composition of mixtures was determined using the mass spectrum computer analysis. Fractions of alkyl benzene, naphthalene and diphenyl derivatives were isolated. The proportions of the individual fractions in the mixtures differed. A predominant fraction was present in each mixture. Solvents (trimethylbenzene, tetramethylbenzene, ethyltoluene, diethylbenzene, isopropylotoulene) which significantly increase health risk due to their suspected neurotoxicity, were identified in each mixture. PMID- 7476151 TI - [A new method for evaluating acute toxicity intragastrically for classification of chemical substances]. AB - The authors reviewed and presented a critical analysis of the literature on defining acute toxicity of chemicals after intragastrical administration by means of traditional methods and new ones such as fixed procedure, up-down, acute-toxic classification. Rules and conditions for the evaluation of acute toxicity by means of these methods were presented as well as the way of defining a median, lethal dose administrated intragastrically for the purpose of chemical classification. PMID- 7476152 TI - [Exposure to mercury and population health. I. Immunotoxicity of mercury]. AB - The authors present large numbers of data on influence of the various mercury compounds on the immune system of humans and laboratory animals. The effect of mercury on the immune system was studied mainly in small rodents intoxicated mostly with HgCl2 or methylmercury. Contrary to a number of these data, analogous observations in humans are rare. Stimulating activity of mercury vapours on the immunological system has been reported in workers at chloralkali plants. So far no data are available concerning influence of the mercury vapours on the resistance of humans to neoplasms and infections. PMID- 7476153 TI - [Stress management: individual differences and determination of situations (review of studies)]. AB - Following the review of studies and their results, the author highlighted the main features of the stress managing process as a basis for taking necessary actions to combat an occupational stress and to ease its possible negative effect. The documented variability of the stress managing provides an opportunity for incorporating the training on how to cope with the stress into the "stress management" programmes at the work place. The relationship between the ways and styles of coping with stress in particular situations and personality traits, significant, however moderate, indicates certain preferences in coping with stress in different situations. Therefore, a certain degree of behavioral "stiffness" as well as an individual approach should be taken into consideration in setting training programmes. The situation-related variability of coping with stress requires distinction between various behavioral patterns in view of occupational situations, namely the kind of stress and the situation control. However, the distinction alone between given behaviours does not provide a sufficient basis for promoting relevant strategies of coping with stress, it is also necessary to evaluate their efficiency, applying various short- and long term efficiency criteria. Strategies which prove to be efficient during a short period of time need not be efficient during a long period of time, and behaviours successful in one situation may fail in another one. PMID- 7476154 TI - [Health effects of unemployment]. AB - The author presents a review of literature dealing with issues concerning unemployment related health effects. Studies of this problem explicitly show that the loss of the job exerts a detrimental health effect. A number of other factors decide about the significance of this relationship. The change from being employed to unemployment affects mostly mental health. First symptoms appear together with the danger of losing the job. Unemployment can evoke apathy, fear or frustration and then it leads to more serious disorders such as neuroses, depression and so forth. It can be also responsible for suicidal attempts. These disorders are usually associated with somatic complaints. Stress evoked by the loss of the job is the major factor affecting the health state of an unemployed person. The loss of income and social status is another important factor. Many authors turn particular attention to a growing intensity of all negative effects of unemployment if it lasts for a long period of time. PMID- 7476155 TI - [On the need for analysis of the occupational health education system]. AB - The paper includes conceptual premises on which the analysis of the occupational health education system is based as well as the outcome of an attempt to perform a comparative study in this area. The authors present some of the features which characterize the education system designed for occupational health physicians. A comparative approach adopted in the analysis (Poland vs some other countries) seems to be the simplest and the most effective way to draw appropriate conclusions. The analysis covered both undergraduate and postgraduate studies, including specialisation in the area of occupational health. Finally, the work highlights theoretical and methodological aspects of further analyses. PMID- 7476156 TI - Pro-sequence-assisted protein folding. AB - Many proteins, including proteases and growth factors, are synthesized as precursors in the form of pre-pro-proteins. Whereas the pre-sequences usually act as signal peptides for transport, the pro-sequences of an increasing number of these proteins have been found to be essential for the correct folding of their associated proteins. In contrast to the action of molecular chaperones, pro sequences appear to catalyse the protein-folding reaction directly. The similarity between the pro-sequence-assisted folding mechanisms of different proteases supports the hypothesis that a common folding mechanism has developed through convergent evolution. Further, the frequent requirement of the pro sequences for both folding and intracellular transport or secretion suggests that these two functionalities are intimately related. PMID- 7476157 TI - The bacterial 'enigma': cracking the code of cell-cell communication. AB - In recent years it has become clear that the production of N-acyl homoserine lactones (N-AHLs) is widespread in Gram-negative bacteria. These molecules act as diffusible chemical communication signals (bacterial pheromones) which regulate diverse physiological processes including bioluminescence, antibiotic production, plasmid conjugal transfer and synthesis of exoenzyme virulence factors in plant and animal pathogens. The paradigm for N-AHL production is in the bioluminescence (lux) phenotype of Photobacterium fischeri (formerly classified as Vibrio fischeri) where the signalling molecule N-(3-oxohexanoyl)-L-homoserine lactone (OHHL) is synthesized by the action of the LuxI protein. OHHL is thought to bind to the LuxR protein, allowing it to act as a positive transcriptional activator in an autoinduction process that physiologically couples cell density (and growth phase) to the expression of the bioluminescence genes. Based on the growing information on LuxI and LuxR homologues in other N-AHL-producing bacterial species such as Erwinia carotovora, Pseudomonas aeruginosa, Yersinia enterocolitica, Agrobacterium tumefaciens and Rhizobium leguminosarum, it seems that analogues of the P. fischeri lux autoinducer sensing system are widely distributed in bacteria. The general physiological function of these simple chemical signalling systems appears to be the modulation of discrete and diverse metabolic processes in concert with cell density. In an evolutionary sense, the elaboration and action of these bacterial pheromones can be viewed as an example of multicellularity in prokaryotic populations. PMID- 7476158 TI - Tracheal colonization factor: a Bordetella pertussis secreted virulence determinant. AB - We report here the identification of a virulence-associated factor, Tcf, (tracheal colonization factor), produced by strains of Bordetella pertussis but not Bordetella parapertussis or Bordetella bronchiseptica. This protein is encoded by the tcfA gene. When a strain of B. pertussis 18323 lacking this protein is used to infect mice with an aerosol challenge, the number of bacteria isolated from the tracheas is decreased 10-fold when compared with the parent 18323. The derived amino acid sequence of tcfA predicts a 68 kDa RGD-containing, proline-rich protein, which after cleavage of a typical prokaryotic signal sequence would be 64 kDa. Amino acid sequence analysis demonstrates that the C terminal 30 kDa of this protein shows 50% identity to the 30 kDa C-terminus of another Bordetella protein, the pertactin precursor. The N-terminal 34 kDa region contains the three amino-acid motif RGD and is 16.5% proline. Coupled in vitro transcription and translation analysis indicates that the tcfA gene product migrates as two bands of approximately 90 kDa. A fusion protein of the N terminal, 34 kDa portion of Tcf to maltose-binding protein migrates, on SDS-PAGE, 30 kDa higher than expected from the combined molecular weights. Polyclonal antisera raised against the unique N-terminal portion of Tcf recognizes 90 kDa and 60 kDa bands in immunoblots of whole-cell lysates of strains of B. pertussis; it does not recognize any protein in whole-cell lysates of B. bronchiseptica or B. parapertussis. Supernatants of cultures of B. pertussis 18323 contain the 60 kDa form of the protein. Southern blot analysis of chromosomal DNA from strains of B. bronchiseptica and B. parapertussis, using a probe derived from tcfA, shows strong hybridization only to B. pertussis DNA. Thus, Tcf appears to be a unique virulence factor of B. pertussis. PMID- 7476159 TI - The chaperone-like protein YerA of Yersinia pseudotuberculosis stabilizes YopE in the cytoplasm but is dispensible for targeting to the secretion loci. AB - The virulence plasmid-encoded YopE cytotoxin of Yersinia pseudotuberculosis is secreted across the bacterial membranes and subsequently translocated into the eukaryotic cell. Translocation of YopE into target cells was recently shown to be polarized and only occurred at the zone of contact between the pathogen and the eukaryotic cell. Immunogold electron microscopy on cryosectioned Y. pseudotuberculosis revealed that YopE is secreted and deposited on the bacterial cell surface when the bacteria are grown in Ca(2+)-depleted media at 37 degrees C. No YopE was detected in the cytoplasm or in the membranes. In yerA mutants which are downregulated for YopE at a post-transcriptional level, the cytotoxin could only be detected in the cytoplasm. The overall recovery of YopE from the yerA mutant strain was, however, considerably lower than from the wild-type strain. yerA had no major effect on the translation of YopE, but was found to stabilize YopE in the cytoplasm. YerA was shown to specifically interact with YopE in the cytoplasm in vivo and this binding also correlated with YopE secretion. Targeting of YopE to the secretion loci as well as translocation of YopE into HeLa cells occurred also in the absence of YerA. Based on our findings, we suggest that YerA by binding to YopE stabilizes and maintains the cytotoxin in a secretion-competent conformation. PMID- 7476160 TI - Promoter selectivity control of Escherichia coli RNA polymerase by ionic strength: differential recognition of osmoregulated promoters by E sigma D and E sigma S holoenzymes. AB - Transcription in vitro of two osmoregulated promoters, for the Escherichia coli osmB and osmY genes, was analysed using two species of RNA polymerase holoenzyme reconstituted from purified core enzyme and either sigma D (sigma 70, the major sigma in exponentially growing cells) or sigma S (sigma 38, the principal sigma at stationary growth phase). Under conditions of low ionic strength, the osmB and osmY promoters were transcribed by both E sigma D and E sigma S. Addition of up to 400 mM potassium glutamate (K glutamate), mimicking the intracellular ionic conditions under hyperosmotic stress, specifically enhanced transcription at these promoters by E sigma S but inhibited that by E sigma D. At similar high concentrations of potassium chloride (KCl), however, initiation at both these promoters was virtually undetectable. These data suggest that the RNA polymerase, E sigma S, itself can sense osmotic stress by responding to changes in intracellular K glutamate concentrations and altering its promoter selectivity in order to recognize certain osmoregulated promoters. PMID- 7476161 TI - Organization of host-inducible transcripts on the symbiotic plasmid of Rhizobium sp. NGR234. AB - In a systematic approach to identify genes involved in the early steps of the legume-Rhizobium symbiosis, we studied transcription patterns of symbiotic plasmid-borne loci. A competitive hybridization procedure was used to identify DNA restriction fragments carrying genes whose expression is enhanced by plant root exudates or by purified flavonoids. Fragments containing induced genes were then located on the physical map of the 500 kb pNGR234a. New inducible loci as well as previously described genes were identified and their time course of induction determined. After initial induction, transcription of loci such as nodABC and the host-specificity genes nodSU decreased to undetectable levels 24 h after incubation with purified flavonoids. In contrast, expression of other loci is detectable only after several hours of induction. Surprisingly, many genes remained transcribed in the nodD1- mutant suggesting the presence of other flavonoid-dependent activators in NGR234. The hsnl region, which is involved in host specificity, was shown to carry several inducible but independently regulated transcripts. Sequencing analysis revealed several open reading frames whose products, based on sequence similarities, may be involved in L-fucose metabolism and its adjunction to the Nod factors. PMID- 7476162 TI - Characterization of IS1221 from Mycoplasma hyorhinis: expression of its putative transposase in Escherichia coli incorporates a ribosomal frameshift mechanism. AB - Seven complete and two partial copies of IS1221 variants from Mycoplasma hyorhinis and Mycoplasma hyopneumoniae characterized to date have established a consensus IS1221 as a 1513 bp element with unique structural characteristics resembling the IS3 family of bacterial insertion sequences. Each IS1221 copy contains highly conserved 28 bp imperfect terminal inverted repeats and three distinctive internal inverted repeats (LIR, RIR and IIR). IIR is located within the coding region of the element and it is proposed that it plays a critical role in the regulation of putative transposase expression. Consensus IS1221 and one particular copy, G1135.2, contain a single long open reading frame (ORF). Two potential initiation codons are present at nucleotide 46 (AUG46) and nucleotide 397 (AUG397) and both are preceded by strong ribosome-binding sites. Both initiation codons can be used efficiently in an Escherichia coli T7 expression system. The LIR has a negative regulatory effect on translation initiation from AUG46. A-1 translational frameshift event is shown to be involved in expression of the IS1221 ORF and results in the production of 20 kDa and 6 kDa truncated proteins from the respective upstream initiation codons of the IS1221 ORF. Base substitution and deletion mutations in sequences resembling characterized motifs in documented examples of translational frameshifting resulted in a significant increase in the full-length products and a corresponding decrease in the truncated products from the IS1221 ORF. In contrast to the usual -1 frameshift regulatory event in the IS3 family, which produces a transframe fusion product as the active transposase, IS1221 may have evolved a high-frequency -1 frameshift mechanism that produces a truncated product from the upstream coding domain and thereby results in the regulated low-level production of the full-length presumptive transposase. PMID- 7476163 TI - The bgl1 gene of Trichoderma reesei QM 9414 encodes an extracellular, cellulose inducible beta-glucosidase involved in cellulase induction by sophorose. AB - We have investigated the effect of disruption of the bgl1-(beta-glucosidase l encoding) gene of Trichoderma reesei on the formation of other beta-glucosidase activities and on the induction of cellulases. To this end the bgl1 locus was disrupted by insertion of the Aspergillus nidulans amdS (acetamidase-encoding) gene. The bgl1-disrupted strain did not produce the 75 kDa extracellular beta glucosidase on cellulose or lactose, but still formed beta-glucosidase activity on glucose, cellobiose, xylan or beta-1,3-glucan, suggesting that the enzyme(s) exhibiting this beta-glucosidase activity is (are) not encoded by bgl1. The cellulase-inducer sophorose induced the bgl1-encoded beta-glucosidase, whereas the remaining beta-glucosidase activity was induced by methyl-beta-D-glucoside. The bgl1-gene product was mainly secreted into the medium, whereas the other beta glucosidase activity was mainly associated with the cells. A bgl1-multicopy strain formed higher amounts of cellulases than the parent strain. Nonsaturating concentrations of sophorose efficiently induced cellobiohydrolase l formation in the bgl1-multicopy strain, but less efficiently in the bgl1-disrupted strain. The multicopy strain and the parent strain were comparably efficient at saturating sophorose concentrations. The beta-glucosidase inhibitor nojirimycin strongly inhibited induction in all strains. These data suggest that the bgl1-encoded beta glucosidase is not identical to the plasma-membrane-bound, constitutive, methyl beta-glucoside inducible beta-glucosidase, but represents an extracellular cellulose-induced enzyme. Both enzymes contribute to rapid induction of cellulases by modifying the inducer sophorose. PMID- 7476164 TI - Mutational analysis of the putative nucleic acid-binding surface of the cold shock domain, CspB, revealed an essential role of aromatic and basic residues in binding of single-stranded DNA containing the Y-box motif. AB - The major cold-shock protein of Bacillus subtilis, CspB, is a member of a protein family widespread among prokaryotes and eukaryotes that share the highly conserved cold-shock domain (CSD). The CSD domain is involved in transcriptional and translational regulation and was shown to bind the Y-box motif, a cis-element that contains the core sequence ATTGG, with high affinity. The three-dimensional structure of CspB, a prototype of this protein family, revealed that this hydrophilic CSD domain creates a surface rich in aromatic and basic amino acids that may act as the nucleic acid-binding site. We have analysed the potential role of conserved aromatic and basic residues in nucleic acid binding by site directed mutagenesis. In gel retardation and ultraviolet cross-linking experiments, the ability of CspB mutants to bind single-stranded oligonucleotides (ssDNA) that contain the Y-box motif was investigated. Single substitutions of three highly conserved phenylalanine residues (Phe-15, Phe-17, Phe-27) by alanine and substitution of one histidine (His-29) by glutamine, all located within the putative RNA-binding sites RNP-1 and RNP-2, abolished the nucleic acid-binding activity of CspB. Conservative substitutions of Phe-15 to tyrosine (F15Y) showed a small increase in binding affinity, whereas separate replacement of Phe-17 and Phe-27 by tyrosine caused a reduction in binding activity. These and other substitutions including the conserved basic residues Lys-7, Lys-13 and Arg-56 as well as the aromatic residues Trp-8 and Phe-30 strongly suggest that CspB uses the side-chains of these amino acids for specific interaction with nucleic acids. Ultraviolet cross-linking experiments for CspB mutants with ssDNA supported the idea of specific CspB/nucleic acid interaction and indicated an essential role for the aromatic and basic residues in this binding. In addition, two-dimensional nuclear magnetic resonance studies with F17A, K13Q, F15Y and F27Y revealed that the mutants have the same overall structure as the wild-type CspB protein. PMID- 7476165 TI - Aminoacyl-tRNA synthetase gene regulation in Bacillus subtilis: induction, repression and growth-rate regulation. AB - The thrS gene in Bacillus subtilis is specifically induced by starvation for threonine and is, in addition, autorepressed by the overproduction of its own gene product, the threonyl-tRNA synthetase. Both methods of regulation employ an antitermination mechanism at a factor-independent transcription terminator that occurs just upstream of the start codon. The effector of the induction mechanism is thought to be the uncharged tRNA(Thr), which has been proposed to base pair in two places with the leader mRNA to induce antitermination. Here we show that the autoregulation by synthetase overproduction is likely to utilize a mechanism similar to that characterized for induction by amino acid starvation, that is by altering the levels of tRNA charging in the cell. We also demonstrate that the base pairing interaction at the two proposed contact points between the tRNA and the leader are necessary but not always sufficient for either form of regulation. Finally, we present evidence that the thrS gene is expressed in direct proportion to the growth rate. This method of regulation is also at the level of antitermination but is independent of the interaction of the tRNA with the leader region. PMID- 7476166 TI - A family of modular type mannuronan C-5-epimerase genes controls alginate structure in Azotobacter vinelandii. AB - The L-guluronic acid residues in the Azotobacter vinelandii polysaccharide alginate originate from a post-polymerization reaction catalysed by the enzyme mannuronan C-5-epimerase (ME). We have previously reported the cloning and expression of an A. vinelandii gene encoding this enzyme, and we show here that the organism encodes at least four other ME genes originating from a common ancestor gene by a complex rearrangement process. The biological function of the corresponding enzymes is probably to catalyse the formation of alginates with a variety of physical properties. This model may explain the origin of the structural variability found in alginates isolated both from prokaryotic and eukaryotic organisms. The A. vinelandii enzymes may also potentially be useful for certain medical and biotechnological applications of this commercially important polysaccharide. PMID- 7476167 TI - Tracheal cytotoxin structural requirements for respiratory epithelial damage in pertussis. AB - The respiratory epithelial pathology of pertussis (whooping cough) can be reproduced by tracheal cytotoxin (TCT), a disaccharide-tetrapeptide released by Bordetella pertussis. TCT is a muramyl peptide, a class of peptidoglycan-derived compounds which have many biological activities including adjuvanticity, somnogenicity, pyrogenicity, and cytotoxicity. The structural requirements for muramyl peptides to produce some of these biological effects have been partially characterized. Using in vitro assays with respiratory epithelial cells and tissue, we have previously determined that the disaccharide moiety of TCT is not involved in toxicity and that the side-chain functional groups of diaminopimelic acid (A2pm) are crucial for toxicity. In this study, we determine the importance of every amino acid, functional group and chiral centre in the peptide portion of TCT. Although lactyl tetrapeptides are the most toxic of the TCT fragments, producing dose-response curves identical to TCT, the smallest analogues of TCT which are active in our assay are of the form X-gamma-(D)-Glu-meso-A2pm, where X may be an amino acid or a blocking group. Within this active substructure, main chain chirality and all functional groups are essential for toxicity. This definition of the core region of TCT indicates that the TCT interaction site is unlike almost all other muramyl peptide interaction sites for which structure activity data are available. PMID- 7476168 TI - Differential effect of dsbA and dsbC mutations on extracellular enzyme secretion in Erwinia chrysanthemi. AB - An Erwinia chrysanthemi gene able to complement an Escherichia coli dsbA mutation has been cloned and sequenced. This gene codes for a periplasmic protein with disulphide isomerase activity that has 69% identity and 94% similarity with the E. coli DsbA protein. An E. chrysanthemi dsbA-uidA fusion mutant has been constructed. dsbA expression seems to be constitutive. This mutant has multiple phenotypes resulting from the absence of disulphide bond formation in periplasmic and secreted proteins. Pectate lyases and the cellulase EGZ are rapidly degraded in the periplasm of the dsbA mutant. E. chrysanthemi synthesizes another periplasmic protein with disulphide isomerase activity, namely DsbC. The dsbC gene introduced on a multicopy plasmid in a dsbA mutant was only partially able to restore EGZ secretion, indicating that even if DsbA and DsbC possess disulphide oxydoreductase activity, they are not completely interchangeable. Moreover, pectate lyases expressed in an E. coli dsbA mutant were very instable but their stability was unaffected in a dsbC mutant. These results indicate that DsbA and DsbC could have different substrate specificities. PMID- 7476169 TI - Gene replacement by homologous recombination in Mycobacterium bovis BCG. AB - Gene replacement by homologous recombination is a powerful tool for fundamental studies of gene function, as well as allowing specific attenuation of pathogens, but has proved difficult to achieve for Mycobacterium tuberculosis. We have used a plasmid-based test system to demonstrate the occurrence of homologous recombination in the tuberculosis vaccine strain Mycobacterium bovis BCG, and we have successfully replaced a target gene in BCG by homologous recombination, using a shuttle plasmid. Specific inactivation of selected genes will facilitate study of virulence factors and drug resistance as well as allowing rational attenuation of M. tuberculosis for the production of new vaccines. PMID- 7476170 TI - Cloning and expression of a murein hydrolase lipoprotein from Escherichia coli. AB - On the basis of the published N-terminal amino acid sequence of the soluble lytic transglycosylase 35 (Slt35) of Escherichia coli, an open reading frame (ORF) was cloned from the 60.8 min region of the E. coli chromosome. The nucleotide sequence of the ORF, containing a putative lipoprotein-processing site, was shown by [3H]-palmitate labelling to encode a lipoprotein with an apparent molecular mass of 36 kDa. A larger protein, presumably the prolipoprotein form, accumulated in the presence of globomycin. Over-expression of the gene, designated mltB (for membrane-bound lytic transglycosylase B), caused a 55-fold increase in murein hydrolase activity in the membrane fraction and resulted in rapid cell lysis. After membrane fractionation by sucrose-density-gradient centrifugation, most of the induced enzyme activity was present in the outer and intermediate membrane fractions. Murein hydrolase activity in the soluble fraction of a homogenate of cells induced for MltB increased with time. This release of enzyme activity into the supernatant could be inhibited by the addition of the serine-protease inhibitor phenylmethylsulphonyl fluoride. It is concluded that the previously isolated Slt35 protein is a proteolytic degradation product of the murein hydrolase lipoprotein MltB. Surprisingly, a deletion in the mltB gene showed no obvious phenotype. PMID- 7476171 TI - Restriction alleviation and modification enhancement by the Rac prophage of Escherichia coli K-12. AB - Bacteriophage lambda encodes an antirestriction function, RaI, which is able to modulate the activity of the Escherichia coli K-12 restriction and modification system, EcoKI. Here we report the characterization of an analogous function, Lar, expressed by E. coli sbcA mutants and the hybrid phage lambda reverse. E. coli sbcA mutants and lambda reverse both express genes of the Rac prophage, and we have located the lar gene immediately downstream of recT in this element. The lar gene has been cloned in an expression plasmid, and a combination of site-directed mutagenesis and labelling of plasmid-encoded proteins has enabled us to identify a number of translational products of lar, the smallest of which is sufficient for restriction alleviation. Lar, like RaI, is able both to alleviate restriction and to enhance modification by EcoKI. Lar, therefore, is functionally similar to RaI and the nucleotide sequences of their genes share 47% identity, indicating a common origin. A comparison of the predicted amino acid sequences of Lar and RaI shows only a 25% identity, but a few short regions do align and may indicate residues important for structure and/or function. PMID- 7476172 TI - Molecular analysis of asmA, a locus identified as the suppressor of OmpF assembly mutants of Escherichia coli K-12. AB - We present the molecular characterization of the asmA gene, whose product is involved in the assembly of outer membrane proteins in Escherichia coli K-12. The asmA locus was initially identified as a site for suppressor mutations of an assembly defective OmpF315. Our data suggest that these suppressor mutations either completely abolish or reduce asmA expression and can be complemented in trans by plasmid clones carrying asmA sequences. The recessive nature of asmA suppressor mutations suggests that the functional AsmA protein participates in inhibiting the assembly of OmpF315 and other mutant OmpFs. As the assembly of wild-type and parental OmpF proteins was not affected by asmA mutations, AsmA must provide an environment refractory only to the assembly of mutant OmpF proteins. However, we cannot completely rule out the possibility that AsmA plays a minor role in the assembly of wild-type and parental OmpF in wild-type cells. The presence of a putative signal sequence within the amino-terminal sequence of AsmA suggests that it is either a periplasmic or an outer membrane protein. This predicted location of AsmA is compatible with its role in the assembly of outer membrane proteins. PMID- 7476173 TI - Role of a potential endoplasmic reticulum retention sequence (RDEL) and the Golgi complex in the cytotonic activity of Escherichia coli heat-labile enterotoxin. AB - Recent experimental evidence indicates that Escherichia coli heat-labile enterotoxin and the closely related cholera toxin gain access to intracellular target substrates through a brefeldin A-sensitive pathway that may involve retrograde transport through the Golgi-endoplasmic reticulum network. The A subunits of both toxins possess a carboxy-terminal tetrapeptide sequence (KDEL in cholera toxin and RDEL in the heat-labile enterotoxins) that is known to mediate the retention of eukaryotic proteins in the endoplasmic reticulum. To investigate the potential role of the RDEL sequence in the toxic activity of the heat-labile enterotoxin we constructed mutant analogues of the toxin containing single substitutions (RDGL and RDEV) or a reversed sequence (LEDR). The single substitutions had little effect on Chinese hamster ovary cell elongation or the ability to stimulate cAMP accumulation in Caco-2 cells. Reversal of the sequence reduced the ability of the toxin to increase cAMP levels in Caco-2 cells by approximately 60% and decreased the ability to elicit elongation of Chinese hamster ovary cells. The effects of the heat-labile enterotoxin were not diminished in a mutant Chinese hamster ovary cell line (V.24.1) that belongs to the End4 complementation group and possesses a temperature-sensitive block in secretion that correlates directly with the disappearance of the Golgi stacks. Collectively, these findings suggest that the brefeldin A-sensitive process involved in intoxication by the heat-labile enterotoxin does not involve RDEL dependent retrograde transport of the A subunit through the Golgi-endoplasmic reticulum complex. The results are more consistent with a model of internalization involving translocation of the A subunit from an endosomal or a trans-Golgi network compartment. PMID- 7476175 TI - The 3'-terminal half of the btuB gene of Citrobacter freundii defines essential residues and supports a topological model for the outer membrane receptor. PMID- 7476174 TI - Characterization of heat-shock response of the marine bacterium Vibrio harveyi. AB - We have investigated heat-shock response in a marine bacterium Vibrio harveyi. We have found that 39 degrees C was the highest temperature at which V. harveyi was able to grow steadily. A shift from 30 degrees C to 39 degrees C caused increased synthesis of at least 10 proteins, as judged by SDS-PAGE, with molecular masses of 90, 70, 58, 41, 31, 27, 22, 15, 14.5 and 14kDa. The 70, 58, 41 and 14.5 kDa proteins were immunologically homologous to DnaK, GroEL, DnaJ and GroES heat shock proteins of Escherichia coli, respectively. V. harveyi GroES protein had a lower molecular mass (14.5 kDa) than E. coli GroES, migrating in SDS-PAGE as 15kDa protein. We showed that a protein of approximately 43 kDa, immunologically reactive with antiserum against E. coli sigma 32 subunit (sigma 32) of RNA polymerase, was induced by heat-shock and co-purified with V. harveyi RNA polymerase. These results suggest that the 43 kDa protein is a heat-shock sigma protein of V. harveyi. Preparation containing the V. harveyi sigma 32 homologue, supplemented with core RNA polymerase of E. coli, was able to transcribe heat shock promoters of E. coli in vitro. PMID- 7476176 TI - Importance of lipooligosaccharide structure in determining gonococcal resistance to hydrophobic antimicrobial agents resulting from the mtr efflux system. AB - Levels of gonococcal resistance to antimicrobial hydrophobic agents (HAs) are controlled by the mtr (multiple transferrable resistance) system, composed of the mtrRCDE genes. The mtrR gene encodes a transcriptional repressor that appears to regulate expression of the upstream and divergent mtrCDE operon. The mtrCDE genes encode membrane proteins analogous to the MexABOprK proteins of Pseudomonas aeruginosa that mediate export of structurally diverse antimicrobial agents. In this study we found that a single base pair deletion in a 13 bp inverted repeat sequence within the mtrR promoter resulted in increased resistance of gonococci to both crystal violet (CV) and erythromycin (ERY) as well as to the more lipophilic non-ionic detergent Triton X-100 (TX-100). However, this cross resistance was contingent on the production of a full-length lipooligosaccharide (LOS) by the recipient strain used in transformation experiments. Introduction of this mutation (mtrR-171) into three chemically distinct deep-rough LOS mutants by transformation resulted in a fourfold increase in resistance to TX-100 compared with a 160-fold increase in an isogenic strain producing a full-length LOS. However, both wild-type and deep-rough LOS strains exhibited an eightfold increase in resistance to CV and ERY as a result of the mtrR-171 mutation. This suggests that gonococci have different LOS structural requirements for mtr mediated resistance to HAs that differ in their lipophilic properties. Evidence is presented that gonococci exclude HAs by an energy-dependent efflux process mediated by the mtr system. PMID- 7476177 TI - Molecular dissection of PapD interaction with PapG reveals two chaperone-binding sites. AB - P pili are composite adhesive fibres that allow uropathogenic Escherichia coli to gain a foothold in the host by binding to receptors present on the uroepithelium via the adhesin PapG. The assembly of P pili requires a periplasmic chaperone, PapD, that has an immunoglobulin-like three-dimensional structure. PapD-subunit complex formation involves a conserved anchoring mechanism in the chaperone cleft and a 'molecular zippering' to the extreme C-terminus of pilus subunits. A chaperone-binding assay was developed using fusions of the C-terminus of PapG to maltose-binding protein (MBP/G fusions) to investigate whether chaperone-subunit complex formation requires additional interactions. PapD bound strongly to an MBP/G fusion containing the C-terminal 140 amino acids of PapG (MBP/G175-314) but only weakly to the MBP/G234-314 fusion containing 81 C-terminal residues, arguing that the region between residues 175-234 contains additional information that is required for strong PapD-PapG interactions. PapD was shown to interact with a PapG C-terminal truncate containing residues 1-198 but not a truncate containing residues 1-145, suggesting the presence of a second, independent PapD interactive site. Four peptides overlapping the second site region were tested for binding to PapD in vitro to further delineate this motif. Only one of the peptides synthesized was recognized by PapD. The MBP/G fusion containing both binding sites formed a tight complex with PapD in vivo and inhibited pilus assembly by preventing chaperone-subunit complex formation. PMID- 7476178 TI - Structural requirements for the glycolipid receptor of human uropathogenic Escherichia coli. AB - The binding of uropathogenic Escherichia coli to the globo series of glycolipids via P pili is a critical step in the infectious process that is mediated by a human-specific PapG adhesin. Three classes of PapG adhesins exist with different binding specificities to Gal alpha 4Gal-containing glycolipids. The structural basis for PapG recognition of the human glycolipid receptor globoside was investigated by using soluble saccharide analogues as inhibitors of bacterial haemagglutination. The minimum binding epitope was confirmed as the Gal alpha 4Gal moiety, but parts of the GalNAc beta and glucose residues, which flank the Gal alpha 4Gal in globoside (GbO4), were also shown to be important for strong binding. Furthermore, the same five hydroxyl groups of Gal alpha 4Gal in globotriasyl ceramide that were recognized by a previously characterized PapG variant were also recognized by the human-specific PapG in binding the GbO4 that dominates in the human kidney. Saccharide analogues that blocked haemagglutination also blocked the adherence of human uropathogenic E. coli to human kidney sections. Knowledge of the molecular details of the PapG-GbO4 interaction will make it possible to design antiadherence therapeutics. PMID- 7476179 TI - Screening of stable proteins in an extreme thermophile, Thermus thermophilus. AB - The leuB gene codes for 3-isopropylmalate dehydrogenase of the leucine biosynthetic pathway in an extreme thermophile, Thermus thermophilus. The leuB gene of the thermophile was replaced with a temperature-sensitive chimeric leuB gene. The resultant transformant was adapted to high temperature, a thermostable mutant strain being obtained. A single base substitution that replaces isoleucine at 93 with leucine was found in the chimeric leuB gene of the thermostable mutant. The resultant amino acid residue coincided with the corresponding residue of the T. thermophilus enzyme. It was confirmed that the mutant enzyme is more stable than the original chimeric enzyme. This system can be used to produce stabilized mutants of other enzymes without structural knowledge of them. PMID- 7476180 TI - Open complex formation by Escherichia coli RNA polymerase: the mechanism of polymerase-induced strand separation of double helical DNA. AB - Escherichia coli RNA polymerase is able to site-specifically melt 12 bp of promoter DNA at temperatures far below those normally associated with DNA melting. Here we consider several models to explain how RNA polymerase destabilizes duplex DNA. One popular model proposes that upon binding to the promoter, RNA polymerase untwists the spacer DNA between the -10 and -35 regions, which results in a destabilization of the -10 region at a TA base step where melting initiates. Promoter untwisting may result, in part, from extensive wrapping of the DNA around RNA polymerase. Formation of the strand-separated open complex appears to be facilitated by specific protein-DNA interactions which occur predominantly on the non-template strand. Recent evidence suggests that these include important contacts with sigma factor region 2.3, which we propose binds the displaced single strand of DNA. PMID- 7476181 TI - The PTR family: a new group of peptide transporters. AB - The transport of peptides into cells is a well-documented biological phenomenon which is accomplished by specific, energy-dependent transporters found in a number of organisms as diverse as bacteria and humans. Until recently, the majority of peptide transporters cloned and characterized were found to be proteins of the ATP-binding cassette (ABC) family. We report the identification of a new family of peptide transporters, which we call the PTR family. This group of proteins, distinct from the ABC-type peptide transporters, was uncovered by sequence analyses of a number of recently discovered peptide transport proteins. Alignment of these proteins demonstrated a high number of identical and similar residues and identified conserved glycosylation and phosphorylation sites, as well as a structural motif unique to this group of proteins. Cluster analysis among the proteins indicated these sequences were indeed related and could be further divided into two subfamilies. A phylogenetic analysis of these new peptide transport sequences, compared to over 50 other peptide and membrane-bound transporters, showed that these proteins comprise a distinct, separate group of proteins. PMID- 7476182 TI - Repair, refold, recycle: how bacteria can deal with spontaneous and environmental damage to proteins. AB - Proteins, like DNA, are subject to various forms of damage that can render them non-functional. Conformational changes and covalent chemical alterations occur spontaneously, and the rates of these reactions can be increased by environmental stresses such as heat, oxidative agents, or changes in pH or osmotic conditions. Although affected proteins can be replaced by de novo biosynthesis, cells- especially those subjected to stress or nutrient limitation--have developed mechanisms which can either restore damaged polypeptides to an active state or remove them. Such mechanisms can spare the biosynthetic capacity of the cell and ensure that the presence of non-functional molecules does not disrupt cell physiology. Three major mechanisms, which operate in bacteria as well as eukaryotic organisms, have been described. First, chaperones not only assist in proper de novo folding of proteins but also provide an important means of restoring activity to conformationally damaged proteins. Second, enzymatic 'repair' systems exist to directly reverse certain forms of protein damage, including proline isomerization, methionine oxidation and the formation of isoaspartyl residues. Finally, proteolysis provides a 'last-resort' means of dealing with abnormal proteins which cannot be repaired. Protein maintenance and repair may be of special importance for bacteria preparing to survive extended periods in stationary phase: both constitutive and induced mechanisms are utilized to permit survival despite greatly reduced protein synthesis. PMID- 7476183 TI - Molecular mechanisms and implications for infection of lipopolysaccharide variation in Neisseria. AB - The lipopolysaccharides of the pathogenic Neisseria species are subject to structural variation owing to a combination of intrinsic changes in lipopolysaccharide (LPS) biosynthesis and external modification of the LPS molecule with sialic acid. This variation appears to control bacterial behaviour by altering their ability to interact with human cells and to evade host immune defences. This interconversion of LPS phenotypes, which is also observed during the natural infection, is probably due to environmental regulation of LPS biosynthesis superimposed on spontaneous changes in the DNA of distinct LPS loci. LPS variation may be a common strategy of mucosal pathogens to colonize and persist within the human host. PMID- 7476184 TI - Cloning, expression and functional analyses of the catabolite control protein CcpA from Bacillus megaterium. AB - A mutant of Bacillus megaterium relieved from catabolite repression has been used to clone ccpA from B. megaterium by complementation. ccpA is the first gene of a presumed operon, in which it is followed by the motA homologue ORF1 and the motB homologue ORF2. The mutation maps in the 3'-terminal region of ccpA, where an in frame duplication of 84 nucleotides located between two 9 bp direct repeats leads to an insertion of 28 amino acids near the C-terminus of CcpA. An in-frame deletion of 501 bp in ccpA exhibits the same phenotype as the 84 bp duplication. Deletion of ORF1 and ORF2 does not yield an apparent phenotype. A single-copy ccpA::lacZ transcriptional fusion is constitutively expressed, independent of whether the growth medium triggers catabolite repression or not. The ccpA mutation leads to relief of catabolite repression exerted by glucose, fructose, mannitol, glucitol and glycerol, whereas only smaller effects were found with ribose, citrate and glutamate. The respective growth rates on these carbon sources are uniformly reduced to a generation time of about 90 min in the ccpA mutant. Catabolite repression of a plasmid-encoded xylA::ccpA fusion is less efficient than that of a xylA::lacZ fusion in the same vector. Furthermore, overproduction of CcpA decreases catabolite repression of a single-copy xylA::lacZ fusion approximately twofold. Thus, overexpression of CcpA may be counterproductive for catabolite repression, supporting the hypothesis that CcpA by itself may not bind sufficiently strongly to the cis-active catabolite responsive element to exert catabolite repression. PMID- 7476185 TI - Characterization of the gene encoding the immunodominant 35 kDa protein of Mycobacterium leprae. AB - Analysis of the interaction between the host immune system and the intracellular parasite Mycobacterium leprae has identified a 35 kDa protein as a dominant antigen. The native 35 kDa protein was purified from the membrane fraction of M. leprae and termed MMPI (major membrane protein I). As the purified protein was not amenable to N-terminal sequencing, partial proteolysis was used to establish the sequences of 21 peptides. A fragment of the 35 kDa protein-encoding gene was amplified by the polymerase chain reaction from M. leprae chromosomal DNA with oligonucleotide primers derived from internal peptide sequences and the whole gene was subsequently isolated from a M. leprae cosmid library. The nucleotide sequence of the gene revealed an open reading frame of 307 amino acids containing most of the peptide sequences derived from the native 35 kDa protein. The calculated subunit mass was 33.7 kDa, but the native protein exists as a multimer of 950 kDa. Database searches revealed no identity between the 35 kDa antigen and known protein sequences. The gene was expressed in Mycobacterium smegmatis under the control of its own promoter or at a higher level using an 'up-regulated' promoter derived from Mycobacterium fortuitum. The gene product reacted with monoclonal antibodies raised to the native protein. Using the bacterial alkaline phosphatase reporter system, we observed that the 35 kDa protein was unable to be exported across the membrane of recombinant M. smegmatis. The 35 kDa protein encoding gene is absent from members of the Mycobacterium tuberculosis complex, but homologous sequences were detected in Mycobacterium avium, Mycobacterium haemophilum and M. smegmatis. The availability of the recombinant 35 kDa protein will permit dissection of both antibody- and T-cell-mediated immune responses in leprosy patients. PMID- 7476186 TI - Novel organization of the site-specific integration and excision recombination functions of the Staphylococcus aureus serotype F virulence-converting phages phi 13 and phi 42. AB - Functions required for site-specific integration and excision of the Staphylococcus aureus serotype F virulence-converting phages phi 13 and phi 42 were localized and characterized. Like other temperate phages, integration of phi 13 and phi 42 sequences was found to require the product of an int gene located close to the phage attP site. Both int genes are almost identical, express proteins possessing characteristic features of the Int (integrase) family of recombinases, but share very little homology with previously described int genes, including those of the serotype B S. aureus phages L54a and phi 11. Nevertheless, all four S. aureus phages share an almost identical short sequence located immediately 5' to these distinct int genes, suggesting a common mechanism of int gene regulation. Upstream from these common sequences, the sequences of phi 13 and phi 42 are quite distinct from each other, and from the corresponding regions of phi 11 and L54a which encode the Xis proteins that are required with Int to mediate site-specific excision of the latter phages. Surprisingly, phi 13 and phi 42 sequences encompassing the attP sites and int genes, but lacking either an adjacent or more distant phage excision protein gene, were sufficient to mediate site-specific excision of integrated phage DNA sequences. PMID- 7476187 TI - Identification of the ligand-binding domain of the surface-located fibrinogen receptor (clumping factor) of Staphylococcus aureus. AB - The ability of Staphylococcus aureus to bind to fibrinogen and fibrin is believed to be an important factor in the initiation of foreign-body and wound infections. Recently, we reported the cloning and sequencing of the gene clfA encoding the fibrinogen receptor (clumping factor, ClfA) of S. aureus strain Newman and showed that the gene product was responsible for the clumping of bacteria in soluble fibrinogen and for the adherence of bacteria to solid-phase fibrinogen. This was confirmed here by showing that antibodies raised against purified Region A inhibited both of these properties. Also, immunofluorescent microscopic analysis of wild-type Newman and a clfA::Tn917 mutant of Newman with anti-ClfA Region A sera confirmed that Region A is exposed on the bacterial cell surface. Furthermore, polystyrene beads coated with the Region A protein formed clumps in soluble fibrinogen showing that the ClfA protein alone is sufficient for the clumping phenotype. Western immunoblotting with anti-ClfA Region A antibodies identified the native ClfA receptor as a 185 kDa protein that was released from the cell wall of S. aureus by lysostaphin treatment. A single extensive ligand binding site was located within Region A of the ClfA protein. Truncated ClfA proteins were expressed in Escherichia coli. Lysates of E. coli and proteins that had been purified by affinity chromatography were tested for (i) their ability to bind fibrinogen in Western ligand blotting experiments, (ii) for their ability to inhibit clumping of bacteria in fibrinogen solution and adherence of bacteria to solid-phase fibrinogen, and (iii) for their ability to neutralize the blocking activity of anti-ClfA Region A antibody. These tests allowed the ligand-binding domain to be localized to a 218-residue segment (residues 332-550) within Region A. PMID- 7476188 TI - The Mycobacterium leprae genome: systematic sequence analysis identifies key catabolic enzymes, ATP-dependent transport systems and a novel polA locus associated with genomic variability. AB - In the framework of the mycobacterial genome sequencing project, a continuous 37,049 bp sequence from the Mycobacterium leprae chromosome has been determined. Computer analysis revealed 10 complete open reading frames, and nine of their products show similarity to known proteins. Seven of these were identified as the enzyme isocitrate lyase, two P-type ATPase cation transporters, two AMP-binding proteins, the ribosomal protein S1, and DNA polymerase I. Interestingly, the polA gene, encoding DNA polymerase, is flanked by two inverted copies of a new class of the M. leprae specific repetitive sequence, RLEP, and this structure resembles a transposable element. A second copy of this element was found at another locus in the genome, but the two copies were not present in equal amounts and could not be found in all isolates of M. leprae. This is the first evidence for genomic variability in the leprosy bacillus and might ultimately be useful for developing a molecular test capable of distinguishing between strains of M. leprae. PMID- 7476189 TI - Unique gene organization of thioredoxin and thioredoxin reductase in Mycobacterium leprae. AB - The thioredoxin system comprising thioredoxin (Trx), thioredoxin reductase (TR) and NADPH operates via redox-active disulphides and provides electrons for a wide variety of different metabolic processes in prokaryotic and eukaryotic cells. Thioredoxin is also a general protein disulphide reductase involved in redox regulation. In bacteria, the Trx and TR proteins previously identified were encoded by separate genes (trxA and trxB). In this study, we report a novel genomic organization of TR and Trx in mycobacteria and show that at least three modes of organization of TR and Trx genes can exist within a single bacterial genus: (i) in the majority of mycobacterial strains the genes coding for TR and Trx are located on separate sites of the genome; (ii) interestingly, in all pathogenic Mycobacterium tuberculosis complex mycobacteria both genes are found on the same locus, overlapping in one nucleotide; (iii) in the pathogen Mycobacterium leprae, TR and Trx are encoded by a single gene. Sequence analysis of the M. leprae gene demonstrated that the N-terminal part of the protein corresponds to TR and the C-terminal part to Trx. A corresponding single protein product of approximately 49 kDa was detected in cell extracts of M. leprae. These findings demonstrate the very unusual phenomenon of a single gene coding for both the substrate (thioredoxin) and the enzyme (thioredoxin reductase), which seems to be unique to M. leprae. PMID- 7476190 TI - Membrane topology and site-specific mutagenesis of Pseudomonas aeruginosa porin OprD. AB - Pseudomonas aeruginosa OprD is a 420-amino-acid protein that facilitates the uptake of basic amino acids, imipenem and gluconate across the outer membrane. OprD was the first specific porin that could be aligned with members of the non specific porin super-family. Utilizing multiple alignments in conjugation with structure predictions and amphipathicity calculations, an OprD-topology model was proposed. Sixteen beta-strands were predicted, connected by short loops at the periplasmic side. The eight external loops were of variable length but tended to be much longer than the periplasmic ones. Polymerase chain reaction (PCR)-based site-specific mutagenesis was performed to delete separately short stretches (4-8 amino acid residues) from each of the predicted external loops. The mutants with deletions in the predicted external loops L1, L2, L5, L6, L7 and L8 were tolerated in both Escherichia coli and P. aeruginosa. The expressed mutant proteins maintained substantial resistance to trypsin treatment in the context of isolated outer membranes. Proteins with deletions in loops L1, L5, L6, L7 and L8 reconstituted similar imipenem supersusceptibility in a P. aeruginosa OprD:: omega background. The L2-deletion mutant only partially reconstituted super susceptibility, suggesting that loop L2 is involved in imipenem binding. These data were generally consistent with the topology model. PMID- 7476191 TI - Negative control of fae (K88) expression by the 'global' regulator Lrp is modulated by the 'local' regulator FaeA and affected by DNA methylation. AB - Expression of the K88 (fae) operon is negatively controlled by the co-operative binding of Lrp and FaeA to the fae regulatory region and is dependent on the methylation status of three GATC sites present in this region. In this paper, we describe the binding of Lrp to a T-rich DNA helix between GATC site I and site II. FaeA stabilized and modified the Lrp binding, thereby extending the Lrp footprint over GATC site I and site III. Methylation of GATC site I prevented the binding of Lrp/FaeA at this site and appeared to be essential for the cells, since mutation of this site into GTTC resulted in a lethal overproduction of K88 fimbriae. Methylation of GATC site II and site III reduced the stability of Lrp/FaeA binding. Moreover, methylation of GATC site III stimulated faeB promoter activity. The plasmid population in cells harbouring multiple copies of a K88 plasmid consisted of two differentially methylated forms. Form A plasmids with a methylated GATC site I and site III and a nonmethylated site II (+,-,+) represented 20% of the population and were responsible for high-level expression. Form B plasmids with a methylated GATC site I and a non-methylated site II and site III (+,-,-) represented 80% of the population and were responsible for low level expression. Apparently, K88 fimbriae expression in vivo is balanced at its maximal possible level by modulation of the methylation status of GATC site III. The ratio (1:4) between these populations is stabilized by a constitutive synthesis of FaeA resulting from the presence of an IS1 insertion upstream of faeA. This IS1 insertion separates the faeA promoter from the FaeB-binding sites, thereby neutralizing the control by FaeB activity on expression of FaeA. Instead, faeA transcription is stimulated by binding of FaeA to the faeA promoter region. PMID- 7476193 TI - Bacillus licheniformis bacitracin-resistance ABC transporter: relationship to mammalian multidrug resistance. AB - The nucleotide sequence of the Bacillus licheniformis bacitracin-resistance locus was determined. The presence of three open reading frames, bcrA, bcrB and bcrC, was revealed. The BcrA protein shares a high degree of homology with the hydrophilic ATP-binding components of the ABC family of transport proteins. The bcrB and bcrC genes were found to encode hydrophobic proteins, which may function as membrane components of the permease. Apart from Bacillus subtilis, these genes also confer resistance upon the Gram-negative Escherichia coli. The presumed function of the Bcr transporter is to remove the bacitracin molecule from its membrane target. In addition to the homology of the nucleotide-binding sites, BcrA protein and mammalian multidrug transporter or P-glycoprotein share collateral detergent sensitivity of resistant cells and possibly the mode of Bcr transport activity within the membrane. The advantage of the resistance phenotype of the Bcr transporter was used to construct deletions within the nucleotide binding protein to determine the importance of various regions in transport. PMID- 7476192 TI - Exploring the Mycoplasma capricolum genome: a minimal cell reveals its physiology. AB - We report on the analysis of 214kb of the parasitic eubacterium Mycoplasma capricolum sequenced by genomic walking techniques. The 287 putative proteins detected to date represent about half of the estimated total number of 500 predicted for this organism. A large fraction of these (75%) can be assigned a likely function as a result of similarity searches. Several important features of the functional organization of this small genome are already apparent. Among these are (i) the expected relatively large number of enzymes involved in metabolic transport and activation, for efficient use of host cell nutrients; (ii) the presence of anabolic enzymes; (iii) the unexpected diversity of enzymes involved in DNA replication and repair; and (iv) a sizeable number of orthologues (82 so far) in Escherichia coli. This survey is beginning to provide a detailed view of how M. capricolum manages to maintain essential cellular processes with a genome much smaller than that of its bacterial relatives. PMID- 7476194 TI - Molecular characterization of the eps gene cluster of Pseudomonas solanacearum and its transcriptional regulation at a single promoter. AB - Production of EPS I, an unusual exopolysaccharide virulence factor of the phytopathogen Pseudomonas solanacearum, requires the 18 kb eps gene cluster. DNA sequence analysis of the first seven genes of eps (epsAPBCDEF), subcellular localization of their products in maxicells, and phoA fusion analysis showed that: (i) epsA, epsB, epsE, and epsF encode exported or membrane-associated proteins probably involved in polymerization and/or export of EPS I; (ii) epsC and epsD encode soluble enzymes probably involved in synthesis of sugar components of EPS I (N-acetylgalactosaminuronic acid and possibly N acetyltrideoxygalactose, respectively); and (iii) epsP probably encodes a phosphatase involved in EPS I production in an unknown way. Non-polar insertional mutagenesis showed that most, if not all, of these eps genes are absolutely required for production of EPS I. Using random eps::lacZ fusions and primer extension we located a transcription start site and promoter upstream of epsA. Analysis of a plasmid with this promoter fused to lacZ showed that a 140 bp regulatory region upstream of the eps transcription start site was sufficient for normal regulation of eps transcription by the multicomponent virulence gene regulatory network of P. solanacearum. Deletion of this eps promoter from a plasmid-borne epsAPBCDE::lacZ fusion reduced its expression 10-fold, indicating that this promoter alone is responsible for regulated transcription of an eps operon composed of at least epsAPBCDE. Analysis of genomic and plasmid-borne eps::lacZ fusions suggested that most remaining eps genes are part of this same operon or, and this is less likely, comprise a second co-ordinately regulated eps operon. PMID- 7476195 TI - rpsL+: a dominant selectable marker for gene replacement in mycobacteria. AB - Molecular genetic manipulations in mycobacteria would benefit from procedures which efficiently select for double-crossover events by homologous recombination. Here we describe a vector-host system for gene replacement in mycobacteria, the utility of which was investigated using functional inactivation of the pyrF gene in Mycobacterium smegmatis as a model. This system is based on the expression of the wild-type rpsL gene coding for ribosomal protein S12 in a streptomycin resistant host. Owing to the absence of a mycobacterial origin the plasmids are unable to replicate autonomously in mycobacteria. The first selection for maintenance of cloned sequences is conferred by the kanamycin-resistance gene. The second simultaneous selection by streptomycin is against maintenance of cloned sequences which contain the gene encoding the streptomycin-sensitive allele of the rpsL gene. By placing the gene for positive selection and that used for negative selection within and outside the target gene of interest, respectively, gene replacement is obtained. A one-step double selection procedure provides a means to distinguish strictly between gene replacement by double crossover versus homologous recombination by single crossover events. The system should have considerable potential for genera or species where single-crossover events or even illegitimate recombination are the predominant recombination mechanisms. It should also be of wide use for the construction of mutants without a selectable phenotype. PMID- 7476196 TI - glmS of Thermus thermophilus HB8: an essential gene for cell-wall synthesis identified immediately upstream of the S-layer gene. AB - A 30 kbp chromosomal region containing the S-layer gene (slpA) from Thermus thermophilus HB8 was cloned from a lambda phage gene library. DNA sequence analysis of the region upstream to the slpA gene revealed the presence of an open reading frame (ORF) which coded for a 604-amino-acid protein highly homologous to the glucosamine-6-P synthases (EC 2.6.1.16) of both prokaryotic and eukaryotic origin. The identification of this ORF as the glucosamine-6-P synthase gene from T. thermophilus (glmSth) has been carried out using three different strategies: (i) complementation of an Escherichia coli glmS mutant; (ii) in vivo insertional inactivation of the gene; and (iii) in vitro synthesis of glucosamine-6-P at 60 degrees C by a cytoplasmic extract of an overproducing E. coli strain. The glmSth gene is transcribed divergently from slpA in a 2.0 kb mRNA which probably also includes a tryptophan tRNA gene (trpTth) identified at its 3' extreme. As the products of both the glmSth and the slpA genes are main components of the cell envelope of T. thermophilus, their unusual clustering in the chromosome could be related to the existence of specific mechanisms for their coordinate expression. PMID- 7476197 TI - tRNA genes and pathogenicity islands: influence on virulence and metabolic properties of uropathogenic Escherichia coli. AB - The uropathogenic Escherichia coli strain 536 (O6:K15:H31) carries two unstable DNA regions on its chromosome which were termed pathogenicity islands (Pais). Both pathogenicity islands, Pai I and Pai II, are incorporated into tRNA specific loci: Pai I is located in the tRNA gene for selenocysteine (selC), and Pai II is integrated in the leucine-specific tRNA locus leuX. Mutant strain 536-21 has lost the two pathogenicity islands together with the intact tRNA genes. While 536 is a virulent strain, 536-21 has lost a number of properties, including in vivo virulence. In previous publications we reported that the genes coding for two haemolysins (hly I, hly II) and P-related fimbria (prf) are located on the Pais. In this paper, we demonstrate that the expression of other gene products influencing metabolic properties in addition to in vivo virulence are strongly dependent on the intact tRNA loci selC and leuX. In order to determine the influence of the two tRNAs on the expression of these properties, the genes selC and leuX were cloned from the genome of strain 536 and then introduced into the mutant 536-21. Our results clearly show that the seleno-cysteine-specific tRNA (tRNA(Sec)) directly influences the ability of the bacteria to grow under anaerobic conditions, because selenocysteine is part of the enzyme formate dehydrogenase (FDH) which is involved in mixed acid fermentation. The rare leucine-specific tRNA5(Leu), encoded by leuX, influences a number if properties including type 1 fimbria production, flagellation and motility, production of enterobactin and serum resistance, and is also necessary for full in vivo virulence. While the tRNA(Sec) is directly involved in the production of FDHs, the leuX specific tRNA5(Leu) appears to influence the expression of various factors through specific transcriptional or translational control mechanisms. PMID- 7476198 TI - SepA, the major extracellular protein of Shigella flexneri: autonomous secretion and involvement in tissue invasion. AB - In addition to Ipa proteins and IcsA, which are involved in entry into epithelial cells and intercellular spread, respectively, Shigella secretes a 110 kDa protein, designated SepA. We report the identification, cloning, and nucleotide sequence determination of the sepA gene, analysis of SepA secretion, and construction and characterization of a sepA mutant. The sepA gene is carried by the virulence plasmid and codes for a 150 kDa precursor. Upon secretion, which does not involve accessory proteins encoded by the virulence plasmid, the precursor is converted to a mature protein of 110 kDa by two cleavages removing an N-terminal signal sequence and a C-terminal fragment. Extensive similarities were detected between the sequence of the first 500 residues of mature SepA and the N-terminal region of IgA1 proteases from Neisseria gonorrhoeae and Haemophilus influenzae, the Tsh haemagglutinin of an avian pathogenic Escherichia coli, and the Hap protein involved in adhesion and penetration of H. influenzae. The C-terminal domain of the SepA precursor, which is not present in the secreted protein, exhibits sequence similarity with pertactin of Bordetella pertussis and the ring-forming protein of Helicobacter mustelae. Construction and phenotypic characterization of a sepA mutant indicated that SepA is required neither for entry into cultured epithelial cells nor for intercellular dissemination. However, in the rabbit ligated ileal loop model, the sepA mutant exhibited an attenuated virulence, which suggests that SepA might play a role in tissue invasion. PMID- 7476199 TI - Identification and characterization of new DNA replication terminators in Bacillus subtilis. AB - A functional DNA replication terminator of Bacillus subtilis contains two overlapping binding sites, A and B, for the replication terminator protein (RTP). A degenerate 17-mer oligonucleotide corresponding to the consensus B site has been used to detect four new terminators in the B. subtilis chromosome, in addition to the previously identified and closely spaced IRI and IRII. All the new terminators lie in the terminus region of the chromosome, on both sides of IRI and IRII, with their positions spanning < 10% of its length. Their DNA sequences are characterized by clearly identifiable A- and B-binding sites. They bind RTP in a manner indistinguishable from IRI, although precise affinities have not been compared. Each new terminator is functional in causing fork arrest when present in a plasmid replicating in B. subtilis. Three of the four were tested for polarity in fork-arrest activity and exhibited the polarity expected. The total of six terminators now identified in B. subtilis have been named TerI TerVI. TerI and TerII correspond to the previously identified IRI and IRII, respectively. The chromosomal orientations of all but one of the terminators (TerIV) have been established and they conform to an arrangement similar to that in Escherichia coli in which two opposed groups of polar terminators provide a replication-fork trap ensuring that the approaching forks meet within a restricted region of the chromosome. The development of a strikingly similar arrangement of terminators in the two organisms, despite the lack of any detectable similarity in their respective DNA terminators and terminator proteins, emphasizes the importance of the replication-fork trap in each case. PMID- 7476200 TI - Characterization of a novel fibronectin-binding surface protein in group A streptococci. AB - Streptococcus pyogenes interacts with host fibronectin via distinct surface components. One of these components is the Sfbl protein (streptococcal fibronectin-binding protein, now specified as class I), an adhesin that represents a protein family with characteristic features. Here we present the complete structure of a novel fibronectin-binding protein of S. pyogenes, designated Sfbll, which is distinct from the previously described Sfbl proteins. The sfbll gene originated from a lambda EMBL3 library of chromosomal DNA from group A streptococcal strain A75 and coded for a 113 kDa protein exhibiting features of membrane-anchored surface proteins of Gram-positive cocci. The expression of biologically active fusion proteins allowed the determination of the location of the fibronectin-binding domain within the C-terminal part of the protein. It consisted of two and a half repeats which share common motifs with fibronectin-binding repeats of other streptococcal and staphylococcal proteins. Purified recombinant fusion protein containing this domain competitively inhibited the binding of fibronectin to the parental S. pyogenes strain. Furthermore, polyclonal antibodies against the binding domain specifically blocked the Sfbll receptor site on the streptococcal surface. No cross-reactivity could be detected between anti-Sfbll antibodies and the sfbl gene product, and vice versa, indicating that the two proteins do not share common immunogenic epitopes. Southern hybridization experiments performed with specific sfbll gene probes revealed the presence of the sfbll gene in more than 55% of 93 streptococcal isolates tested. The majority of the strains also harboured the sfbl gene, and 86% carried at least one of the two sfb genes. PMID- 7476201 TI - GvpCs with reduced numbers of repeating sequence elements bind to and strengthen cyanobacterial gas vesicles. AB - We have previously shown that the gas-vesicle protein GvpC is present on the outer surface of the gas vesicle, can be reversibly removed and rebound to the surface, and increases the critical collapse pressure of the gas vesicle. The GvpC molecule, which contains five partially conserved repeats of 33 amino acids (33-RR) sandwiched between 18 N-terminal and 10 C-terminal amino acids, is present in a ratio of 1:25 with the GvpA molecule, which forms the ribs of the gas vesicle. By using recombinant techniques we have now made modified versions of GvpC that contain only the first two, three or four of the 33-amino-acid repeats. All of these proteins bind to and strengthen gas vesicles that have been stripped of their native GvpC. Recombinant proteins containing three or four repeats bind in amounts that give the same ratio of 33-RR:GvpA (i.e. 1:5) as the native protein, and they restore much of the strength of the gas vesicle; the protein containing only two repeats binds at a lower ratio (1:7.7), however, and restores less of the strength. Ancestral proteins with only two, three or four of the 33-amino-acid repeats would have been functional in strengthening the gas vesicle but the progressive increase in number of repeats would have provided strength with increased efficiency. PMID- 7476202 TI - The stationary-phase sigma factor sigma S (RpoS) is required for a sustained acid tolerance response in virulent Salmonella typhimurium. AB - The acid tolerance response (ATR) of log-phase Salmonella typhimurium is induced by acid exposures below pH 4.5 and will protect cells against more extreme acid. Two systems are evident: a transiently induced system dependent on the iron regulator Fur that provides a moderate degree of acid tolerance and a more effective sustained ATR that requires the alternate sigma factor sigma S encoded by rpoS. Differences between the acid responses of virulent S. typhimurium and the attenuated laboratory strain LT2 were attributed to disparate levels of RpoS caused by different translational starts. The sustained ATR includes seven newly identified acid shock proteins (ASPs) that are dependent upon sigma S for their synthesis. It is predicted that one or more of these ASPs is essential for the sustained system. The sustained ATR also provided cross-protection to a variety of other environmental stresses (heat, H2O2 and osmolarity); however, adaptation to the other stresses did not provide significant acid tolerance. Therefore, in addition to starvation, acid shock serves as an important signal for inducing general stress resistance. Consistent with this model, sigma S proved to be induced by acid shock. Our results also revealed a connection between the transient and sustained ATR systems. Mutations in the regulator atbR are known to cause the overproduction of ten proteins, of which one or more can suppress the acid tolerance defect of an rpoS mutant. One member of the AtbR regulon, designated atrB, was found to be co-regulated by sigma S and AtbR. Both regulators had a negative effect on atrB expression. The results suggest AtrB serves as a link between the sustained and transient ATR systems. When sigma S concentrations are low, a compensatory increase in AtrB is required to engage the transiently induced, RpoS-independent system of acid tolerance. Results also suggest different acid-sensitive targets occur in log-phase versus stationary phase cells. PMID- 7476203 TI - PhoP/PhoQ transcriptional repression of Salmonella typhimurium invasion genes: evidence for a role in protein secretion. AB - Previously, the PhoP-repressed locus prgH was identified as important for signalling epithelial cells to endocytose Salmonella typhimurium. Characterization of prgH revealed that it is an operon of four genes encoding polypeptides of 392 (prgH), 80 (prgI), 101 (prgJ) and 252 amino acid residues (prgK). Synthesis of the 2.6 kb prgHIJK transcript was repressed in bacteria that activate PhoP/PhoQ, indicating that PhoP/PhoQ regulates prgHIJK by transcriptional repression. The prgI, prgJ and prgK predicted gene products were similar to Shigella flexneri and Yersinia enterocolitica proteins required for secretion of Ipa and Yop virulence factors. Analysis of the culture supernatants from wild-type S. typhimurium demonstrated that at least 25 polypeptides larger than 14 kDa could be detected. In contrast, prgH1::TnphoA, phoP-constitutive and hil-deletion mutants had significant defects in their supernatant protein profiles. The invasion and supernatant protein profile defects of the prgH1::TnphoA mutant were both complemented by a 5.1 kb plasmid that included prgHIJK. These results suggest that PhoP/PhoQ regulates extracellular transport of proteins by transcriptional repression of secretion determinants and that secreted proteins may be involved in signalling epithelial cells to endocytose bacteria. PMID- 7476204 TI - A multiple site-specific DNA-inversion model for the control of Omp1 phase and antigenic variation in Dichelobacter nodosus. AB - The molecular cloning and sequence analysis of four structurally variant linked genes (omp1A,B,C,D) that encode the major outer membrane protein of Dichelobacter nodosus strain VCS1001 are described. The isolation of rearranged copies of omp1A and omp1B, and the identification in the 5' regions of all four genes of short cross-over-site sequences that were similar to the Din family of cross-over-site sequences, suggested that site-specific DNA inversion was involved in omp1 rearrangement. Evidence for site-specific inversion of the 497 bp DNA fragment, which was located between the divergently orientated omp1A and omp1B genes, and which contained the promoter and 5' coding sequence of Omp1, was obtained by polymerase chain reaction-mediated amplification of inverted forms of these genes. However, to account for all of the omp1 gene copies cloned in this study, a more widespread inversion phenomenon must be involved in the rearrangement of these genes and a model for multiple site-specific DNA inversions at the omp1 locus is described. In this model the four structurally variant omp1 genes can be assembled from one of four structurally variant C-terminal coding regions and a conserved N-terminal coding region and can be expressed from a single promoter. It is postulated that this genetic capability endows D. nodosus with the ability to switch the antigenic specificity of one of its major surface proteins. PMID- 7476206 TI - The role of cysteine residues in the transport of mercuric ions by the Tn501 MerT and MerP mercury-resistance proteins. AB - Each cysteine residue in the MerT and MerP polypeptides of bacterial transposon Tn501 was replaced by serine, and the mercury-resistance phenotypes of the mutants were determined in Escherichia coli. Cys-24 and Cys-25 in the first transmembrane region of MerT were essential for transport of mercuric ions through the cytoplasmic membrane, and mutations Cys-76-Ser, Cys-82-Ser or Gly-38 Asp in MerT or Cys-36-Ser in MerP all reduced transport and resistance. Deletion of the merP gene slightly reduced mercuric ion resistance and transport, whereas a Cys-33-Ser mutation in MerP appears to block transport of mercuric ions by MerT. The effects of deleting merP on mutations in merT were tested. The 116 amino-acid MerT protein is sufficient for mercuric ion transport across the cytoplasmic membrane. PMID- 7476205 TI - Characterization and regulation of the expression of FatB, an iron transport protein encoded by the pJM1 virulence plasmid. AB - The pJM1-encoded genes fatDCBA are essential for iron acquisition via the siderophore anguibactin. Sequence analysis indicated that the open reading frame corresponding to the fatB gene possesses domains that are characteristic of periplasmic proteins that bind the ferric siderophore. In this work, a monospecific antiserum against an oligopeptide containing the last 27 amino acids of the carboxy-terminal region from this open reading frame was used to demonstrate that fatB encodes a 35 kDa protein that is essential for iron transport. By using this antibody we were able to demonstrate that expression of the fatB gene is negatively regulated by the Fur protein at high iron concentrations. Conversely, its expression was positively regulated by the combined action of the AngR protein and products of the TAF region. FatB, the product of the fatB gene, is isolated with the membrane fraction. In accordance with these findings is the fact that the first 23 amino acid residues of this protein have the properties of a lipoprotein signal sequence. The lipoprotein nature of FatB is supported by the fact that treatment of Vibrio anguillarum cells with globomycin, an inhibitor of the lipoprotein signal peptidase, results in the accumulation of a 38 kDa proFatB precursor protein. PMID- 7476207 TI - A new RNA polymerase sigma factor, sigma F, is required for the late stages of morphological differentiation in Streptomyces spp. AB - A gene (sigF) encoding a new sigma factor was isolated from Streptomyces aureofaciens using a degenerate oligonucleotide probe designed from the GLI(KDNE)A motif lying within the well-conserved region 2.2 of the eubacterial sigma 70 family. Homologues were present in other Streptomyces spp., and that of the genetically well studied Streptomyces coelicolor A3(2) was also cloned. The nucleotide sequences of the two sigF genes were determined and shown to encode primary translation products of 287 (S. coelicolor) and 295 (S. aureofaciens) amino acid residues, both showing greatest similarity to sigma B of Bacillus subtilis. However, while sigma B is involved in stationary-phase gene expression and in the general stress response in B. subtilis, sigma F affects morphological differentiation in Streptomyces. Disruption of sigF did not affect vegetative growth but did cause a whi mutant phenotype. Microscopic examination showed that the sigF mutant produced spores that were smaller and deformed compared with those of the wild type, that the spore walls were thinner and sensitive to detergents and that in sigF mutant spores the chromosome failed to condense. sigma F is proposed to control the late stages of spore development in Streptomyces. PMID- 7476208 TI - Co-ordinate, temperature-sensitive regulation of the three Yersinia enterocolitica flagellin genes. AB - Yersinia enterocolitica cells, when cultured at 30 degrees C or below, are flagellated and motile. Cells cultured at 37 degrees C or above lack flagella and are non-motile. To identify flagellin genes that are a target of this temperature dependent regulation, a library of Y. enterocolitica genomic inserts in a phage lambda vector was probed with the Salmonella typhimurium fliC (flagellin) gene. A DNA fragment subcloned from a recombinant phage which hybridizes with the probe complements a non-motile S. typhimurium fliC-fljB- (flagellin-minus) mutant. DNA sequence analysis shows that Y. enterocolitica contains three tandem flagellin genes, designated fleA, fleB and fleC. All three genes are co-ordinately transcribed at low, but not high, temperature from fliA-dependent (sigma F) promoters. Flagellin transcription arrests rapidly after upshift to 37 degrees C (host temperature). In contrast, flagellin transcription resumes only after several generations when cells cultured at 37 degrees C are downshifted to 28 degrees C. PMID- 7476209 TI - A physical genome map of the Burkholderia cepacia type strain. AB - Burkholderia cepacia (basonym Pseudomonas cepacia), the type species of the new genus Burkholderia, is of interest, not only because of its broad catabolic capacity and its ability to antagonize soil-borne plant pathogens, but also because of its causative role in infections in man, which are particularly evident in patients with cystic fibrosis. A physical map of the 8.1 Mb genome of the B. cepacia type-strain ATCC 25416 was constructed by applying two-dimensional pulsed-field gel electrophoresis techniques. Placed onto the macrorestriction map were 38 SpeI, 11 SwaI, 11 PacI, 11 PmeI and six I-CeuI sites, resulting in an average resolution of 105 kbp. Random single-hit linearization by irradiation and restriction mapping uncovered the presence of four circular replicons of 3.65 Mb, 3.17 Mb, 1.07 Mb and 200 kbp in size. The largest replicon harbours four rrn operons while the other two Megabase-size replicons each contain a single rrn operon, suggesting that the genome has three chromosomes and a large plasmid. Within the beta subdivision of proteobacteria, the existence of multiple replicons is not confined to B. cepacia. The phylogenetically related species Burkholderia glumae, Burkholderia pickettii, Burkholderia solanacearum, Alcaligenes eutrophus and the so far unassigned Pseudomonas glathei were also found to harbour more than one Megabase-size replicon. PMID- 7476210 TI - Cloning and molecular characterization of two genes encoding adhesion proteins involved in Trichomonas vaginalis cytoadherence. AB - Cytoadherence to the vaginal epithelium is a critical step in infection by the eukaryotic flagellate Trichomonas vaginalis. Four trichomonad surface proteins (AP65, AP51, AP33 and AP23) mediate cytoadherence. The cDNA encoding the AP65 adhesin was isolated from a phagemid cDNA expression library by screening with antiserum and monoclonal antibody (mAb) raised against the purified trichomonad AP65 protein. Two clones, F11.2 and F11.5, coded for immuno-crossreactive recombinant proteins that possessed functional properties equal to the T. vaginalis AP65 adhesin. Analysis of full-length sequences corresponding to the F11.2 and F11.5 cDNAs revealed that both contained 1701-base open reading frames (ORFs) that encoded proteins of 63 281 daltons and 83 087 daltons, respectively. Comparison of the full-length sequences showed 87% identity at the nucleotide level and 91% identity at the protein level. Restriction-enzyme mapping and Southern analysis reaffirmed the distinctness of the F11.2 and F11.5 cDNAs, indicating that two different AP65 genes (now called ap65-1 and ap65-2) are present in the T. vaginalis genome in at least two copies each. Northern analysis detected high levels of transcript of approximately 1.8 kb for both ap65-1 and ap65-2 genes in trichomonads grown only in high-iron medium, confirming the transcriptional regulation of adhesin synthesis by iron. Homology searches revealed significant similarity (38% amino acid identity and 54% nucleotide identity) to malic enzymes. However, purified malic enzyme and mAb to AP65 crossreactive with malic enzyme neither inhibited cytoadherence of T. vaginalis to host cells nor prevented binding of the trichomonad AP65 to HeLa cells in a ligand assay. PMID- 7476211 TI - Long stretches of short tandem repeats are present in the largest replicons of the Archaea Haloferax mediterranei and Haloferax volcanii and could be involved in replicon partitioning. AB - We report the presence of long stretches of tandem repeats in the genome of the halophilic Archaea Haloferax mediterranei and Haloferax volcanii. A 30 bp sequence with dyad symmetry (including 5 bp inverted repeats) was repeated in tandem, interspersed with 33-39 bp unique sequences. This structure extends for long stretches--1.4 kb at one location in H. mediterranei chromosome and about 3 kb in the H. volcanii chromosome. The tandem repeats (designated TREPs) show a similar distribution in both organisms, appearing once or twice in the H. volcanii and H. mediterranei chromosomes, and once in the largest, probably essential megaplasmid of each organism but not in the smaller replicons. Sequencing of the structures in both H. volcanii replicons revealed an extremely high sequence conservation in both replicons within the species, as well as in the different organisms. Homologous sequences have also been found in other more distantly related halophilic members of the Archaea. Transformation of H. volcanii with a recombinant plasmid containing a 1.1 kb fragment of the TREPs produced significant alterations in the host cells, particularly in terms of cell viability. The introduction of extra copies of TREPs within the vector significantly alters the distribution of the genome among the daughter cells, as observed by DAPI staining. Although the precise biological role cannot be completely ascertained, all the data conform with the tandem repeats being involved in replicon partitioning in halobacteria. PMID- 7476212 TI - Cloning and characterization of GPD2, a second gene encoding sn-glycerol 3 phosphate dehydrogenase (NAD+) in Saccharomyces cerevisiae, and its comparison with GPD1. AB - We have cloned and characterized a homologue of the previously isolated GPD1 gene, encoding sn-glycerol 3-phosphate dehydrogenase (NAD+) in Saccharomyces cerevisiae. This second gene, called GPD2, encodes a protein of 384 amino acids that shares 69% sequence identity with GPD1. Like GPD1 it has an amino-terminal extension of unknown function. GPD2 is located on chromosome VII and cross hybridizes with GPD1 at chromosome IV as well as with an unknown homologue at chromosome XV. Disruption of the GPD2 gene did not reveal any observable phenotypic effects, whereas overexpression resulted in a slight, but significant, increase of GPD enzyme activity in wild-type cells. Analysis of gene transcription by a CAT-reporter gene fused to the GPD promoters revealed decreased transcriptional activity of the GPD2 promoter in cells grown on nonfermentable as opposed to fermentable carbon sources, and no induction in cells exposed to high osmolarity or heat shock. Similar analysis of GPD1 demonstrated an 8-17-fold higher basal level of transcription compared to GPD2. Furthermore, such analysis revealed that the GPD1 promoter was induced by increased osmolarity essentially independent of the type of stress solute used, the level of GPD1 transcription being increased about sevenfold in cells growing at 1.4 M NaCl. PMID- 7476213 TI - Great lives at Manguinhos. Henrique de Beaurepaire Rohan Aragao. 21.12.1879 25.02.1956. PMID- 7476214 TI - The epidemiological importance of Triatoma nigromaculata (Stal, 1859) colonizing human dwellings of the Venezuelan Andes. PMID- 7476215 TI - Living and avoiding malaria infection in endemic areas of the Amazon Basin. PMID- 7476216 TI - Observations on some avian Coccidia (Apicomplexa: Eimeriidae) in Amazonian Brazil. AB - Oocysts of Eimeria porphyrulae n.sp. are described in faeces of Porphyrula martinica (Aves: Gruiformes: Rallidae). They are ellipsoidal to oval, 22.4 x 17.7 (20.0-23.7 x 16.2-18.7) microns, shape-index (length/width) 1.3. Oocyst wall about 1.25 microns thick, colourless, with two layers: inner one prominently striated. Micropyle and sub-micropylar granule present: no oocyst residuum. Sporocysts 17.5 x 9.0 (17.0-19.0 x 8.0-10.0) microns, shape-index 1.9, with inconspicuous Stieda/sub-Stieda bodies. Sporocyst residuum of scattered granules, sometimes a compact mass: sporozoites with two refractile bodies. Eimeria crypturelli n.sp. is described in faeces of Crypturellus soui (Tinamiformes: Tinamidae). Oocysts ellipsoidal-oval, 20.75 x 14.5 (17.5-25.0 x 11.25-21.25) microns, shape-index 1.4. Oocyst wall about 1.25 microns thick and bi-layered: inner layer faintly striated. Micropyle present, with oocyst residuum immediately below: single polar body rarely present. Sporocysts 13.0 x 7.5 (12.5-13.75 x 7.5 8.1) microns, shape-index 1.7, with a Stieda body but seemingly no sub-Stieda. Sporocyst residuum compact: sporozoites with two refractile bodies. Isospora cacici n.sp. is recorded from faeces of Cacicus cela cela (Passeriformes: Icteridae). Oocysts subspherical-spherical, 26.5 x 23.7 (22.5-27.5 x 20.0-26.2) microns, shape-index 1.1. Wall a single, colourless layer about 1.5 microns thick. No micropyle or oocyst residuum: 1-2 polar bodies. Sporocysts ellipsoidal, 17.7 x 12.5 (17.5-18.75 x 11.25-13.75) microns, shape-index 1.4, with pronounced Stieda/sub-Stieda bodies: residuum compact and sporozoites with two refractile bodies. Isospora thraupis n.sp. is described from faeces of Thraupis palmarum melanoptera (Passeriformes: Thraupidae).(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7476217 TI - ELISA for the detection of human leptospirosis. PMID- 7476218 TI - Detection of Trypanosoma cruzi and Leishmania using the polymerase chain reaction. PMID- 7476219 TI - Prevalence of antibodies against an immunodominant region of the HIV-1 envelope glycoprotein gp41 in sera from HIV-1 infected individuals in Brazil. PMID- 7476220 TI - Electrocardiographic alteration among first degree relatives with serologic evidence of Trypanosoma cruzi infection. A sibship study. AB - To analyze whether electrocardiographic alterations (ECGA) in patients with antibodies to Trypanosoma cruzi showed a pattern of familial aggregation, a sample of 379 young adults (166 men and 213 women) distributed in sibships, were assessed for the presence of anti-T. cruzi antibodies, and subjected to a complete clinical examination and a standard resting electrocardiogram (ECG). Positive T. cruzi serology was detected in 165 individuals, 48 of them showing an abnormal ECG (overall prevalence 29%). One hundred and eleven seropositive individuals were distributed in 45 sibships, each of them constituted by more than one seropositive sib, with ECGA being present in 34 out of these patients. Seropositive subjects with ECGA were detected in 27 sibships. Since the index case within each sibship is counted exactly once, affected individuals selected at random as propositi were extracted to calculate the prevalence of ECGA among first degree relatives of probands. Abnormal ECGs were recorded in 7 out of 45 sibs yielding a prevalence that did not differ from estimations registered in the general population or seropositive sibs. Data from the present sample show no familial aggregation for the occurrence of ECGA in patients with T. cruzi. infection. PMID- 7476221 TI - Hepatitis B vaccination in leprosy patients. PMID- 7476222 TI - Morphological aspects of the myocarditis and myositis in Calomys callosus experimentally infected with Trypanosoma cruzi: fibrogenesis and spontaneous regression of fibrosis. AB - Calomys callosus a wild rodent, is a natural host of Trypanosoma cruzi. Twelve C. callosus were infected with 10(5) trypomastigotes of the F strain (a myotropic strain) of T. cruzi. Parasitemia decreased on the 21st day becoming negative around the 40th day of infection. All animals survived but had positive parasitological tests, until the end of the experiment. The infected animals developed severe inflammation in the myocardium and skeletal muscle. This process was pronounced from the 26th to the 30th day and gradually subsided from the 50th day becoming absent or residual on the 64th day after infection. Collagen was identified by the picro Sirius red method. Fibrogenesis developed early, but regression of fibrosis occurred between the 50th and 64th day. Ultrastructural study disclosed a predominance of macrophages and fibroblasts in the inflammatory infiltrates, with small numbers of lymphocytes. Macrophages had active phagocytosis and showed points of contact with altered muscle cells. Different degrees of matrix expansion were present, with granular and fibrillar deposits and collagen bundles. These alterations subsided by the 64th days. Macrophages seem to be the main immune effector cell in the C. callosus model of infection with T. cruzi. The mechanisms involved in the rapid fibrogenesis and its regression deserve further investigation. PMID- 7476224 TI - Effects of the spore-endotoxin complex of a strain of Bacillus thuringiensis serovar morrisoni upon Triatoma vitticeps (Hemiptera: Reduviidae) under laboratory conditions. PMID- 7476223 TI - Ultrastructural study on experimental infection of rotavirus in a murine heterologous model. AB - Viral replication, histopathological and ultrastructural changes were observed for a period of nine days in the small intestine of suckling mice infected with a simian rotavirus (SA11). Samples taken from duodenum, jejunum and ileum were prepared for light microscopy, transmission and scanning electron microscopy analysis. Histopathologic effect could be detected within 8 hr post-infection, when only a few altered cells were observed. Damage was extensive after 16 hr post-infection, showing swollen enterocytes and reduced and irregularly oriented microvilli at intestinal villi tips. Virus particles were detected at 16 and 48 hr post-infection, budding from the viroplasm into the rough endoplasmic reticulum cisternae in ileum enterocytes. Clear evidence of viral replication, observed by electron microscopy was not described before in heterologous murine models. Regeneration of the intestinal villi began at the third day post infection. Despite some differences observed in clinical symptoms and microscopic analysis of homologous and heterologous rotavirus infections, we concluded that mechanisms of heterologous rotavirus infection in mice follow similar patterns to those observed in the homologous models. PMID- 7476225 TI - Host-induced morphological changes of Schistosoma mansoni Sambon, 1907 male worms. AB - In order to evaluate the permissiveness of Nectomys squamipes to Schistosoma mansoni and the influence of the albino mice on the morphological aspects of adult worms derived from a population isolated from N. squamipes, the morphology of adult S. mansoni Sambon, 1907 male worms was studied using a digital image analyser (MOP VIDEOPLAN) and light microscopy. Their sources were as follows: (1) recovered from the wild rodent N. squamipes Brants naturally infected from Sumidouro, RJ, Brazil; (2) recovered from albino mice experimentally infected with the strain derived from N. squamipes; (3) recovered after the isolation of a strain derived from aboriginal human infections in Sumidouro. Worms recovered from N. squamipes (group 1) showed body length and distance between suckers significantly bigger than those of the specimens maintained in mice (groups 2 and 3). The number of testes in group 1 was statistically less than that of groups 2 and 3. Group 2 strains which were maintained in mice, presented the length of the worms as the only significant different character. Data show that: (1) N. squamipes is a more suitable host for the development of S. mansoni when compared to the albino mice; (2) a strain of S. mansoni isolated from a natural host undergoes morphological changes after its passage in the white mouse. PMID- 7476226 TI - Observation on the host parasitoid interaction between Periplaneta americana, the American cockroach and Tetrastichus hagenowii, an oothecal parasitoid. AB - Investigations were carried out on the host parasitoid interaction between Periplaneta americana, the American cockroach and Tetrastichus hagenowii, an oothecal parasitoid. This gregarious female parasitoid infected and or oviposited in only one host and caused 100% mortality of the infected host. However, increase in parasitoid density decreased the progeny production and influenced the sex ratio. The progenies produced were male biased. When host preference was tested by offering oothecae of different species of cockroaches. T. hagenowii showed a predilection towards the oothecae of P. americana, suggesting its host specificity. PMID- 7476227 TI - Endemic leishmaniasis in Brazil: future implications. PMID- 7476228 TI - Ecoepidemiological aspects of American cutaneous leishmaniasis in the state of Sao Paulo, Brazil. AB - The aim of this study is to review some of the ecoepidemiological aspects of american cutaneous leishmaniasis (ACL) in the State of Sao Paulo, Brazil. During the first half of this century ACL occurred in Sao Paulo, predominantly on the bank of the Tiete River, where there were railroad constructions and there was inevitable contact between workers and forested areas. Man's activities resulted in a drastic reduction of the forested regions of the State and molded the present landscape found in Sao Paulo, which brought a gradual change in the epidemiology of ACL during this century. Currently ACL can be considered as an endemic disease. Nowadays, ACL is found in different regions of Sao Paulo State, and is no longer limited to the bank of the Tiete River. The disease occurs in all age groups and sexes. Lack of knowledge about wild reservoir hosts of Leishmania (V.) braziliensis has simulated speculation about the possible role played by domestic animals (dogs and equines). Man's activities also favoured Lutzomyia intermedia a sandfly species which can clearly thrive in changed environments. L.(V.) braziliensis continues to be transmitted, even after decades of forest destruction in Sao Paulo. PMID- 7476229 TI - The dermal leishmaniases of Brazil, with special reference to the eco epidemiology of the disease in Amazonia. AB - Six species of Leishmania are at present known to cause cutaneous and/or mucocutaneous leishmaniasis in Brazil, and they are all to be found in the Amazon region of this country. The eco-epidemiology of each is discussed, with the observation that the Amazonian leishmaniases are all zoonoses, with their source in silvatic mammals and phlebotomine sandfly vectors. With man's destruction of the natural forest in southern Brazil, some sandfly species have survived by adapting to a peridomestic or domiciliary habitat in rural areas. Some domestic animals, such as dogs and equines are seemingly now involved in the epidemiology of the disease. No such process has yet been reported in the Amazon region, but may well take place with the continuing devastation of its forest. PMID- 7476230 TI - American cutaneous leishmaniasis in Pernambuco, Brazil: eco-epidemiological aspects in 'Zona da Mata' region. AB - American cutaneous leishmaniasis (ACL) has been increasing in Pernambuco, thus becoming an important problem for Public Health. The incindence is predominant in the region called 'Zona da Mata', in the east of this state. This region corresponds geographically to the primitive area of the Atlantic forest. In order to characterize the eco-epidemiology expression of ACL in this region, two localities situated in the municipalities of Amaraji e Cortes have been selected by the criterion of higher incindence of human cases. Five stocks of patients were characterized and identified on the basis of enzyme profiles as a new variant of Leishmania (V.) braziliensis. A survey of wild and domestic animals was carried out by means of a parasitological and serological diagnosis. Through the analysis of the spleen and liver imprints, were detected amastigotes compatible with Leishmania in five Nectomys s. squamipes, five Bolomys 1. pixuna, two Rattus r. alexandrinus and one Rattus r. frugivorus. For two years we carried out monthly sandflies captures using CDC light traps as well as manual captures. Lutzomyia whitmani was predominant, which accounted for 97.4% of the total. These data indicate a strong evidence on the vector and the potential reservoirs of L. braziliensis in this region. PMID- 7476231 TI - Vectors control importance on leishmaniasis transmission. AB - We reviewed the control of transmission of leishmaniasis regarding chemotherapy, reservoirs elimination, vaccination and insect control through the use of chemical insecticides. We also discussed complementary measures like monitoring traps, impregnated bednets and curtains, repelents, pheromones, biological control, etc. A cost comparison of insecticide interventions through the use of products belonging to the four main chemical groups was also done, comparing together conventional formulations versus a slow-release insecticide developed by the Nucleo de Pesquisas de Produtos Naturais, Universidade Federal do Rio de Janeiro. We finally did recommendations on the situation that would justify an insecticide intervention to control sandflies. PMID- 7476232 TI - Sand fly vectorial ecology in the State of Sao Paulo. AB - Ecological aspect of sand fly distribution in the State of Sao Paulo, Brazil are described. The main man-biting species are Lutzomyia whitmani, Lu.pessoai, Lu.intermedia, Lu.migonei and Lu.fischeri. Their primary habitat is the forest but latter three of the above species are also encountered in domiciliary environment. Sylvatic species such as Lu.flaviscutellata bite man only rarely and Psychodopygus ayrozai seems to be more anthropophilic. The survival of sand flies in the residual forest and in cultivated areas where man has nearly destroyed the forest almost completely is analyzed. Over the last ten years the incidence of human American cutaneous leishmaniasis (ACL) has been increasing: human cases occurring within several municipalities in which there is overlapping with the distribution of domiciliary Lu.intermedia. New ACL microfoci are appearing in the State of Sao Paulo and these call for further study. PMID- 7476233 TI - The chemical control of vectors of leishmaniasis. PMID- 7476234 TI - Use of molecular probes and PCR for detection and typing of Leishmania--a mini review. AB - The use of molecular tools to detect and type Leishmania species in humans, reservoirs or sandflies has been pursued using different approaches. The polymerase chain reaction provided sensitivity to case this task, since the use of hybridization procedures alone employing specific probes is hampered due to the low detection limit. In this report, we describe the different molecular targets used in our laboratory, aiming at the detection and specific typing of these protozoa. Different kits based on hybridization assays and PCR amplification using kinetoplast and nuclear targets are described and the results obtained from their use are reported. PMID- 7476235 TI - Taxonomy of the genus Leishmania: present and future trends and their implications. AB - The application of different taxonomic methods (Cladistic, Evolutionary Taxonomy and Numerical Taxonomy) to the taxonomy of the Genus Leishmania are reviewed. The major groupings of the most recent classifications obtained using the cladistical approach agree with the major divisions of previous classifications which used traditional taxonomy (Evolutionary Taxonomy). The advantage of the cladistical approach is that it produces cladograms whose branches indicate more accurately levels of relationships between the different taxa. Numerical Taxonomy is useful for identification but not as good as the cladistical approach for classification. The ancient division of this monophyletic genus into two major evolutionary lines supports the use of the subgeneric names Leishmania and Viannia. PMID- 7476236 TI - Exploitation of parasite derived antigen in therapeutic success of human cutaneous leishmaniasis in Brazil. AB - In a complete study in 25 patients with American cutaneous leishmaniasis, caused by Leishmania braziliensis complex, immunotherapeutic efficacy of parasite derived antigen (94-67 KD) has been compared to antimonial therapy. Additionally, to delineate the mechanism of therapeutic success, microscopical features of immune response in active lesions and healed or non-healed lesions following therapy were analyzed. The results showed that cure rates in immunotherapy and chemotherapy were equal (> 83%). The immunohistochemical changes in two therapeutic groups were also largely similar. The analysis of humoral and cellular immune response suggest that appropriate stimulation of T helper cells in the lesion site, in association with one or more cytokines, play a key role in the healing process. PMID- 7476237 TI - Personal experience with diagnostic and therapeutic aspects of human Leishmania (Viannia) braziliensis in Tres Bracos. AB - Diagnostic and therapeutic aspects of human infection with Leishmania (Viannia) braziliensis found in the littoral forest of the State of Bahia are reviewed. There is pressing need for alternative cheap oral drug therapy. PMID- 7476238 TI - The Brazilian Leishmaniasis Control Program. AB - The control methods used in Brazil for both cutaneous and visceral leishmaniasis by the Ministry of Health's National Health Foundation (Fundacao Nacional de Saude) are presented. Data referring to the programme between 1980 and 1991 is presented. There was an increase in the total number of notified cases of cutaneous leishmaniasis until 1987 and from this date onwards the number has remained between 23,000 and 26,000 per year. There were significantly fewer cases of visceral leishmaniasis during the same period and until 1991 the greatest number of cases reported in one year was 2,511 in 1985. The overall picture is that it is difficult to evaluate to what extent the control methods employed have reduced the number of cases of either form of the disease. It is suggested that there is an urgent need for a closer approximation of research and health workers to review the actual control strategies and to define procedures capable of assessing accurately their impact. PMID- 7476240 TI - Familiarity and attention: does what we know affect what we notice? AB - Previous work on the object and word superiority effects has demonstrated that activation from stored representations can facilitate identification of items in a visual display. We predicted that activation of this sort might exogenously attract visual attention toward items that have stored representations. To test this prediction, we presented a familiar (word) and an unfamiliar (nonword) item simultaneously at unpredictable locations, and after varying delays, moved one of the stimuli. In accord with our prediction, at the shortest intervals subjects were more efficient at discriminating motion of the familiar item. Control data demonstrated that this advantage was due to a competitive interaction and not to the familiarity of the items per se. PMID- 7476239 TI - Recognition of familiar and unfamiliar melodies in normal aging and Alzheimer's disease. AB - We tested normal young and elderly adults and elderly Alzheimer's disease (AD) patients on recognition memory for tunes. In Experiment 1, AD patients and age matched controls received a study list and an old/new recognition test of highly familiar, traditional tunes, followed by a study list and test of novel tunes. The controls performed better than did the AD patients. The controls showed the "mirror effect" of increased hits and reduced false alarms for traditional versus novel tunes, whereas the patients false-alarmed as often to traditional tunes as to novel tunes. Experiment 2 compared young adults and healthy elderly persons using a similar design. Performance was lower in the elderly group, but both younger and older subjects showed the mirror effect. Experiment 3 produced confusion between preexperimental familiarity and intraexperimental familiarity by mixing traditional and novel tunes in the study lists and tests. Here, the subjects in both age groups resembled the patients of Experiment 1 in failing to show the mirror effect. Older subjects again performed more poorly, and they differed qualitatively from younger subjects in setting stricter criteria for more nameable tunes. Distinguishing different sources of global familiarity is a factor in tune recognition, and the data suggest that this type of source monitoring is impaired in AD and involves different strategies in younger and older adults. PMID- 7476241 TI - Stimulus-independent thought depends on central executive resources. AB - Stimulus-independent thoughts (SITs) are streams of thoughts and images unrelated to immediate sensory input. Four experiments examined the contribution of aspects of working memory to production of SITs. In Experiments 1 and 2, interventions that were targeted on, respectively, phonological and visuospatial components of working memory both interfered with production of SITs, but there was evidence that these tasks also made demands on central executive resources. Experiments 3 and 4 specifically examined the hypothesis that production of SITs and control of nonproceduralized tasks both depend on central executive resources, and so should show mutual interference. In Experiment 3, prior practice on pursuit rotor and memory tasks reduced the interference with SITs from concurrent task performance. In Experiment 4, randomness within a task involving random-number generation was less when SITs were being produced concurrently than it was when they were not. The results suggest that production of SITs depends on central executive resources. PMID- 7476242 TI - Tilted letters and tilted words: a possible role for principal axes in visual word recognition. AB - Subjects made lexical decisions to columnar letter strings in which every letter was either upright or tilted 90 degrees clockwise as if the whole letter string had been rotated from the horizontal. Lexical decisions were faster in the latter case. The advantage for the tilted format was also found when all strings were presented in aLtErNaTiNg CaSe or in uppercase, so this advantage cannot be due to preservation of the tilted words' global shape. The cost for the upright-letter format increased with the number of letters in the columnar strings. These data suggest that words recognition may involve shape description or position coding relative to a reference frame based on the principal axis of the letter string. PMID- 7476243 TI - The word frequency effect in recognition memory versus repetition priming. AB - The role of word frequency in recognition memory and repetition priming was investigated by using a manipulation of attention. In Experiment 1, the lexical decision task produced greater repetition priming for low-frequency words than for high-frequency words following either the attended or the unattended study condition. The recognition memory test, on the other hand, showed a low-frequency word advantage only following the attended study condition. Furthermore, this advantage was limited to the measure of recognition memory based on conscious recollection of the study episode. In Experiment 2, a speeded recognition memory test replicated the pattern obtained with the unspeeded recognition memory test in Experiment 1. These results argue against the view that the word frequency effects in recognition memory and repetition priming have the same origin. Instead, the results suggest that the word frequency effect in recognition memory has its locus in conscious recollection. PMID- 7476244 TI - Effects of memory load in a word-naming task: five failures to replicate. AB - Paap and Noel's (1991) recent demonstration that increased memory load facilitates naming of low-frequency irregular words has been taken as strong support for dual-route theories of word naming (e.g., Coltheart, 1978). Others, however, (Jared, personal communication, March 1993; Strain, personal communication, April 1993) have been unable to replicate this effect. In the present research, five different attempts were made to determine the crucial experimental conditions required for successful replication of the Paap and Noel findings. None of these experiments produced results at all similar to those reported by Paap and Noel. A number of explanations for these failures to replicate, in terms of individual differences between Paap and Noel's subjects and the present subjects, were evaluated. Most of these explanations were not supported by the data. The present results call into question the generalizability of the Paap and Noel findings and, hence, the strength of support they provide for dual-route theories. PMID- 7476245 TI - Uncertainty in estimating distances from memory. AB - Magnitude estimations involving spatial characteristics, such as distance, typically show a compressive function when estimates are made from memory. In particular, as the magnitude of a property grows larger and larger, estimates become more and more inaccurate, with increasing underestimates of the actual magnitude. Previous theories have attempted to explain this difference by supposing that magnitude estimation was accomplished through a reperceptual process, in which the errors of perception are magnified, or a transformation process, in which the memory trace undergoes a consistent alteration toward a more schematic form. The present experiments present evidence in support of an uncertainty hypothesis. When subjects are uncertain of the actual value of a distance, they are forced to guess on the basis of the mean distance they encountered, because they are unable to retrieve the information accurately. When they can retrieve the information, they are more certain and their estimates are more accurate. This hypothesis was also extended to integrative conditions in which the subjects were presented with the stimulus display in a piecemeal fashion. In these cases, distance estimates were derived by combining spatial representations. This method of presentation caused distance estimates to become less accurate. PMID- 7476246 TI - The recollective experience of cross-modality confusion errors. AB - There are at least two qualitatively different types of subjective experience that can accompany accurate recognition memory. Sometimes recognition is accompanied by conscious recollection of the learning episode (i.e., it is remembered), and in other cases it is not (i.e., it is simply known that the item is old). In the present study, we assessed the subjective experience that accompanies cross-modality confusion errors (misidentifying words as pictures) by measuring the extent to which subjects claimed to remember versus know that the item was presented as a picture. The results of two experiments demonstrate that cross-modality confusion errors are often accompanied by conscious recollection, although not to the same extent as accurate memories. The findings also show that there is considerable overlap in the recollective experience that accompanies accurate and erroneous memories. Finally, the results support the contention that recollective experience cannot be directly inferred from test performance. PMID- 7476247 TI - A multinomial modeling analysis of the recognition-failure paradigm. AB - The recognition-failure paradigm has received much theoretical consideration, especially the Tulving-Wiseman function and its exceptions. We show that the Tulving-Wiseman function does a poor job of accounting for the data, both when its fit is measured with a model-based, goodness-of-fit statistic and when a logically equivalent reformulation of the function is compared with data. We then present a simple multinomial model based on retrieval-independence theory that is capable of measuring storage and retrieval processes in recognition failure. The model is used to conduct a meta-analysis of the recognition-failure paradigm, and shows that violations of the Tulving-Wiseman function occur under conditions in which weak storage is coupled with strong retrieval. In addition, if storage and retrieval are assumed to be positively correlated across conditions, the model produces a theoretically motivated, alternative equation to the Tulving-Wiseman function that provides a virtually identical fit to the data. PMID- 7476248 TI - Developing TODAM: three models for serial-order information. AB - TODAM2, a theory of distributed associative memory, shows how item and associative information can be considered special cases of serial-order information. Consequently, it is important to get the right model for serial order information. Here, we analyze and compare three distributed-memory models for serial-order information that use TODAM's convolution-correlation formalism. These models are the chaining model, the chunking model, and a new model, the power-set model. The chaining model associates each item with its predecessor; the chunking model uses multiple convolutions and n-grams to form chunks; and the power-set model interassociates all items in a set in a particular way to form a chunk. The models are compared in terms of their performance on seven basic tests of serial-order information--namely, serial recall, backward recall, recall of missing items, sequential probe tests, positional probe tests, serial-to-paired associate transfer, and item recognition. The strengths and weaknesses of each model are discussed. PMID- 7476249 TI - Naive theories and causal deduction. AB - Evidence is presented that implicates two factors in deductive reasoning about causality. The factors are alternative causes and disabling conditions (factors that prevent effects from occurring in the presence of viable causes). A causal analysis is presented in which these factors impact on judgments concerning causal necessity and sufficiency, which in turn determine deductive entailment relations. In Experiment 1, these factors were found to impact causal deductive judgments more strongly than did logical form. In Experiment 2, causal deductive judgments were found to vary as a function of familiarity with a particular causal relationship: The more familiar the causal relationship, the less willing reasoners were to accept conclusions based on them. PMID- 7476250 TI - Effects of sampling context on the expressive language of children and adolescents with mental retardation. AB - This study was designed to determine whether conversational samples of language should be supplemented with samples obtained in narrative, or storytelling, contexts when evaluating the expressive language skills of individuals with mental retardation. Language samples were elicited in both conversational and narrative contexts from 16 school-age individuals with mental retardation and 16 typically developing children matched to them on MA. An analysis of the samples demonstrated that both groups produced more syntactically complex language in narration than in conversation, whereas they were more talkative in conversation than in narration. These results suggest that conversational and narrative contexts should be used together when evaluating the expressive language performance of individuals with mental retardation. PMID- 7476251 TI - Supporting the free exercise of religion in the group home context. AB - Ways were delineated in which group home employees can support individuals living in the home in the active expression of faith, both inside and outside the group home environment, and at the same time protect the freedom of others in the home who choose not to be involved in religious activities. After briefly reviewing some assumptions underlying the 1992 AAMR definition of mental retardation and describing the capability of religious expression for individuals with cognitive impairment, we presented several guidelines to assist group home staff members. PMID- 7476252 TI - Validity and reliability of a Five Dimensional Life Satisfaction Index. AB - Validity of the Five Dimensional Life Satisfaction Index (Hawkins, Eklund, Kim, & Greene, 1994), which was developed to measure life satisfaction as it applies to adults with mental retardation, was examined. The index was administered to 48 adults with moderate mental retardation. Results showed (a) alternate forms reliability and internal consistency, (b) convergent and discriminant validity, and (c) construct validity. PMID- 7476253 TI - Analyzing Behavior State and Learning Environments: application in instructional settings. AB - An evaluation of the effects of the ABLE (Analyzing Behavior State and Learning Environments) model was presented. Eight teachers received training on observing behavior state and relevant environmental variables; analyzing potential effects of nutrition and medications; identifying strategies for individual students; and participating in a process to generate classroom-based intervention strategies based on their observations. Results showed that teachers were able to reliably implement the model, which produced significant increases in occurrences of the preferred alert and active states and significant decreases in nonpreferred states, such as sleep, drowse, daze, and crying/agitated. Additional findings indicated that the teachers were satisfied with the training procedures and found the model to be relevant and appropriate for students with profound mental retardation. Case studies were briefly presented. PMID- 7476254 TI - Voluntary transformation from an institutionally based to a community-based service system. AB - The transformation of a large, private, not-for-profit, church-affiliated provider of residential services from an institutionally based to community-based service system was described. Closure of a 200-person ICF/MR facility was discussed. Factors influencing the decision to close the institution as well as the guidelines used in effecting the transformation were described. Finally, data were presented indicating that consumer and staff satisfaction and judgments of program quality remained high during the period of transformation. PMID- 7476255 TI - Probabilities of the development of ambulation for mobile children with moderate to profound mental retardation. AB - Development of ambulation was studied among 1,744 children, ages 3 through 10 years, with moderate to profound mental retardation. The sample was limited to children with some degree of mobility at initial testing. Evaluations over 6 years indicated that improvements in basic skills, although not probable, do occur far past the normative age for development of speech and ambulation and across all levels of mental retardation. The probability of improvement declined with age. Degree of mental retardation and severity of motor impairment had the expected negative effects on initial skill level and probability of improvement. Limiting the sample to subjects who were mobile minimized the effect of mortality. PMID- 7476256 TI - A humanist's legacy: Burton Blatt and the origins of the disability rights movement. PMID- 7476257 TI - Contributions and limitations of human services. PMID- 7476258 TI - Managed care and the future of primary care for adults with mental retardation. PMID- 7476259 TI - Keeping facilitated communication in perspective. PMID- 7476260 TI - Comment on "Review of the role of two antilibidinal drugs in the treatment of sex offenders with mental retardation". PMID- 7476263 TI - Detection of germ cell mutagenicity of trophosphamide by the spermatid micronucleus test in the rat. AB - The effects of the antineoplastic drug trophosphamide (TP) on male rat germ cells were examined with the spermatid micronucleus test (SMNT). We used the microdissection technique in order to isolate stage I of the seminiferous epithelial cycle, where the cells that have just passed the meiotic divisions can be found. Micronuclei (MN) were scored at different time-points after TP treatment at dose levels of 25 and 50 mg/kg. An induction of MN was detected in cells exposed at preleptotene (18 and 19 days) and late pachytene (3 days), as well as at the diplotene-diakinesis stage (1 day). The dose-response for MN induction was linear at all time intervals studied, except for 18 days time point. The highest frequency of MN (5.20 +/- 0.57/1000 spermatids) could be found with the lower TP dose at 18 days, corresponding to exposed preleptotene spermatocytes and reflecting S-dependent clastogenicity. While a significant increase in MN could only be detected in exposed preleptotene spermatocytes with the lower TP dose, the higher dose level also induced MN significantly in late pachytene and the diplotene-diakinesis stage. DNA flow cytometry at 18 days showed cytotoxicity of TP to exposed primary spermatocytes at pachytene, but no cytotoxicity to the preleptotene spermatocytes that exhibited a significant MN induction. The results show that the SMNT using the stage-I-specific examination of the rat seminiferous epithelium can detect the germ cell mutagenicity of TP and gives further evidence of the usefulness of this technique in the testing of chemicals for genotoxic effects in male germ cells. PMID- 7476261 TI - Cytogenetic and immunofluorescence analysis of benzo[a]pyrene-DNA adduct formation and chromosome damage in larval brain neuroblasts of Drosophila melanogaster. AB - Recently we have evaluated the relationship between benzo[a]-pyrene(BaP)-DNA adducts, determined by 32P-postlabelling, and clone frequencies in the somatic mutation and recombination test (SMART) in Drosophila melanogaster. Following that study we proceeded to characterise further the mechanism of induction of genetic damage in vivo by BaP in Drosophila by cytogenetic analysis of larval brain neuroblasts. Third stage larvae were treated with 4 and 10 mM BaP for 24, 48 or 72 h. In all cases, the larvae were killed 72 h after the beginning of treatment, entailing 48, 24 or 0 h post-treatment recovery in BaP-free medium, respectively. At the end of the treatment the following data were collected: (i) the types and levels of chromosome aberrations in neuroblast metaphase and anaphase nuclei; (ii) the distribution and level of BaP-DNA adducts, revealed by indirect immunofluorescence in neuroblast nuclei using an anti-(BaP-DNA) antibody. The results indicate that BaP induces chromosome breaks, deletions and exchanges in this system. In particular, chromosome exchanges decrease as the post-treatment recovery time increases, and the dynamics of breaks and deletions appear to be inversely related to those of the exchanges. This suggests that exchanges may require few preconditions to occur and are thus expressed soon after treatment. Chromosome breaks and deletions could require multiple single events before the actual damage is expressed (even some cell divisions away from the end of treatment). The immunofluorescence analysis suggests that BaP-DNA adducts are more abundant in the heterochromatin of the neuroblast nuclei.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7476262 TI - Early G1 in the male rat meiotic cell cycle is hypersensitive to N-methyl-N nitrosourea-induced micronucleus formation. AB - The effects of the known carcinogenic and teratogenic agent N-methyl-N nitrosourea (MNU) were studied on male rat meiosis. To examine possible cell cycle delay, an immunohistochemical technique based on 5-bromo-2'-deoxyuridine (BrdU) labelling of S-phase cells was developed. BrdU tablets were implanted subcutaneously in adult male rats. A single i.p. injection of 10 mg/kg of MNU was given simultaneously. After 16-22 days, preparations of stage I of the seminiferous epithelium were made and stained immunohistochemically using anti BrdU antibodies. MNU did not cause any significant meiotic delay, but did cause a slight non-significant reduction of the percentages of BrdU-labelled step 1 spermatids at 18 days (80%) compared to controls (95%). In addition, the induction of meiotic micronuclei was studied after short (1-3 days: late meiotic stages) and long (16-22 days: early spermatocytes and B spermatogonia) exposure times. The peak induction occurs between 21 and 20 days, indicating that the M-G1 transition or the very beginning of G1 of the cell cycle of primary spermatocytes are the most sensitive stages of the action of MNU. The number of step 1 spermatids decreased dramatically in animals treated for 22 days, denoting a highly toxic effect on type-B spermatogonia. No unscheduled DNA synthesis was detected in any meiotic stage of spermatogenesis by using this BrdU labelling method. The results indicate that the spermatid micronucleus test based on microdissection of seminiferous tubules can accurately point out the most sensitive stage for chemically induced clastogenesis. Moreover, the BrdU immunohistochemical application enables the simultaneous study of cell cycle kinetics.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7476264 TI - Tannic acid is not mutagenic in germ cells but weakly genotoxic in somatic cells of Drosophila melanogaster. AB - Tannic acid (TA) was tested for genotoxic activity in three different assays (1 3) in Drosophila melanogaster by feeding of larvae or adult flies. TA did not induce sex-linked recessive lethals (1) nor sex-chromosome loss, mosaicism or non disjunction (2) in male germ cells. In the wing somatic mutation and recombination test (SMART) (3) TA was found to be toxic for larvae of the high bioactivation cross and produced a weak positive response. These results suggest that this compound, when administered orally to larvae or adults of D. melanogaster, is not mutagenic and clastogenic in male germ cells, but weakly genotoxic in somatic cells of the wing imaginal disk. PMID- 7476265 TI - Genotoxicity testing: current practices and strategies used by the pharmaceutical industry. AB - Current guidelines and recommendations for genotoxicity testing of pharmaceuticals are disparate, both in terms of the most appropriate tests to use and the protocols to follow. Recent attempts have been made to standardise genotoxicity testing procedures, coinciding with the current review of the OECD guidelines and the International Conference on Harmonisation of Technical Requirements for Registration of Pharmaceuticals for Human Use (ICH). However, as with other aspects of non-clinical safety assessment of pharmaceuticals, guidelines have been prepared by evaluation of general chemical data due to the lack of specific information on pharmaceuticals. To address this, a project was undertaken to collect and collate information specifically pertaining to the genotoxicity testing of pharmaceuticals in order to obtain a clear understanding of international strategy and procedures in the pharmaceutical industry. It is clear that the practices and regional variations are strongly influenced by national guidelines and do not necessarily follow companies' preferences. However, there is a surprising amount of variation in approach between companies on some issues. This is evident in how companies define a genotoxin. This ranges from a positive result in an in vivo assay as indicative of a genotoxin (43%) to any positive result in vitro or in vivo (30%). Indeed many companies (particularly in Japan) will terminate development on the strength of a clear positive result in an Ames test. There is much debate within the ICH process concerning tests to detect gene mutations in mammalian cells as part of a primary test battery. This survey shows that in general, the pharmaceuticals industry has severe doubts about these assays. Thirty-seven (78%) of the 47 participating pharmaceutical companies include an in vitro test to detect gene mutation in mammalian cells as part of their routine test battery. The HPRT test using Chinese hamster cells has the most widespread use, although there is only limited use of such tests in Japan. Compound development has been affected by the results of such tests, but usually only in terms of clarification of equivocal results in other genotoxicity tests in the test battery. The majority (63%) of companies do not support its use as a primary regulatory requirement, and 83% do not consider the mouse lymphoma assay (L5178Y) an acceptable replacement for in vitro mammalian cytogenetics. In conclusion, this survey has provided valuable information on the current modus operandi of the international pharmaceutical industry for consideration in current harmonisation initiatives. PMID- 7476266 TI - An optimal, generalized sampling time of 30 +/- 6 h after double dosing in the mouse peripheral blood micronucleus test. AB - Double dosing and single sampling seems to be the simplest and most reliable method for detecting clastogens in the mouse peripheral blood micronucleus test. Optimal sampling times after double dosing are studied here. Eleven clastogens [water soluble: colchicine, cyclophosphamide, cytosine arabinoside, 5-fluoro-2' deoxyuridine, 5-fluorouracil (5-FU), 6-mercaptopurine, methotrexate (MTX), mitomycin C; water insoluble: N-2-acetylaminofluorene, diethyl sulfate, 7,12 dimethyl-benz[a]anthracene (DMBA)] were given i.p. twice to MS/Ae mice and peripheral blood samples were collected at 6 h intervals starting 24 h after the second treatment. Samples collected at 30 +/- 6 h were nearly optimal for all 11 chemicals. When DMBA, 5-FU and MTX were given, elevated micronucleus frequencies tended to last longer relative to those induced by direct-acting chemicals. Since samples collected 30 +/- 6 h after the second treatment with these three chemicals showed almost the same micronucleus frequencies observed in later samples, the time 30 +/- 6 h could be applied for these long-acting chemicals. Micronucleated reticulocytes persist in the peripheral blood (PB) longer than micronucleated polychromatic erythrocytes are retained in the bone marrow, which thus provides a simple, effective and generalized protocol for the mouse short term PB micronucleus test, namely double dosing and sampling 30 +/- 6 h after the second dose. When one sample time has to be selected, 30 h after the second treatment is recommended. PMID- 7476267 TI - Lack of uniformity in the mutational spectra of chlorohydroxyfuranones in Salmonella typhimurium strain TA100. AB - The mutational specificity of three chlorohydroxyfuranones found in chlorinated drinking water, 3-chloro-4-(dichloromethyl)-5-hydroxy-2(5H)-furanone (MX), 3 chloro-4(chloromethyl)-5-hydroxy-2(5H)-furanone (CMCF) and 3,4-dichloro-5-hydroxy 2(5H)-furanone (mucochloric acid, MCA), was examined in Salmonella typhimurium strain TA100. DNA colony-hybridization of TA100 revertants showed that MX and CMCF both induced predominantly G:C-->T:A transversions (87 and 75% of total, respectively) with a 3:1 preference for the second position of the hisG46 (CCC) target codon. By contrast, MCA produced primarily G:C-->A:T transitions (66% of the total) with a 4:1 preference for the second position of the CCC codon. The mutational specificity of MCA is consistent with the idea that chloroacetaldehyde, a degradation product of MCA, is responsible for the observed mutations. The chemical mechanism by which either MX or CMCF induces G:C-->T:A transversions remains unknown. PMID- 7476268 TI - Structural requirements for mutagenicity of flavonoids upon nitrosation. A structure-activity study. AB - A wealth of promutagens can damage DNA provided metabolic/chemical reactions take place before an ultimate mutagen is formed. Nitrosation reactions are amongst those chemical reactions which may take place to render some chemical classes of promutagens as ultimate mutagens. Flavonoids are amongst chemicals which can be rendered mutagenic upon nitrosation. In this study, 22 flavonoids were tested in the Ames assay for their mutagenicity upon nitrosation and the respective structural requirements for nitrosation-dependent mutagenicity were established. Nitrosatable chemicals present in the diet may play a role in the aetiology of gastric cancer and flavonoids are amongst the common molecules present in a variety of food items. Flavonoids such as quercetin and catechin were predicted to be non-mutagenic upon nitrosation by the CASE methodology and were shown in this study to be strong nitrosatable mutagens. PMID- 7476269 TI - The micronucleus assay in human exfoliated urothelial cells: effect of smoking. AB - Micronucleated cell rates were examined in exfoliated urothelial cells of 73 healthy donors not occupationally exposed to mutagens or aneugens. Micronucleated cell levels averaged 0.54 +/- 0.68% in the entire population: they reached 1.09% in smokers, 0.95% in ex-smokers and 0.24% in non-smokers. Among variation factors evaluated during this study, only smoking had a significant effect on micronucleated cell rates (P = 0.007) whereas age and sex had no effect (P = 0.101 and P = 0.918). No significant difference was observed between micronucleated cell rates of smokers and ex-smokers, suggesting that smoking could generate clones of basal micronucleated cells in urothelial tissues. PMID- 7476270 TI - Use of the SOS chromotest, the Ames-fluctuation test and the newt micronucleus test to study the genotoxicity of four trihalomethanes. AB - Three short-term assays (the SOS chromotest, the Ames-fluctuation test and the newt micronucleus test) were carried out to evaluate the genotoxicity of four trihalomethanes (chloroform, bromodichloromethane, chlorodibromomethane and bromoform). With the SOS chromotest, all the chemicals studied except chloroform were found to induce primary DNA damage in Escherichia coli PQ37. In the Ames fluctuation test, only bromoform showed mutagenic activity on Salmonella typhimurium strain TA100. The newt micronucleus assay detected a clastogenic effect on the peripheral blood erythrocytes of Pleurodeles waltl larvae for bromodichloromethane and bromoform. It appeared that the presence of bromine substituent(s) generally led to significant genotoxic activity. Moreover, the use of the metabolic system significantly increased the genotoxicity of the brominated trihalomethanes in the SOS chromotest. Unlike previous investigations in which the SOS chromotest was always the least interesting assay, this study exhibited the good efficiency of this in vitro test on E.coli for the detection of trihalomethanes with bromine substituents. PMID- 7476271 TI - Genotoxicity of 17 gyrase- and four mammalian topoisomerase II-poisons in prokaryotic and eukaryotic test systems. AB - The genotoxic potency of certain classes of topoisomerase II poisons is correlated with their affinity to the topoisomerase protein rather than with the presence of 'classical' structural alerts for DNA reactivity: bacterial topoisomerase II poisons (specifically named gyrase inhibitors) are highly genotoxic in prokaryotic systems; mammalian topoisomerase II poisons are potent mutagens/clastogens in eukaryotic systems. Studies with bacterial, lower eukaryotic and mammalian genotoxicity tests were performed to draw structure activity conclusions and address risk-benefit considerations for the class of quinolone gyrase inhibitors. All 17 gyrase inhibitors investigated in this study showed genotoxic activity in Salmonella typhimurium strain TA102 and the SOS test. The genotoxic and the toxic activities increased in a highly parallel fashion from the parent compounds, nalidixic acid and oxolinic acid, to the new generation fluoroquinolones. Generally, the most potent fluoroquinolones also show clear-cut positive effects in eukaryotic test systems, although at concentrations 100-1000-fold higher than those effective in bacteria and also 100 1000-fold higher than the minimal genotoxic concentrations of antitumour topoisomerase II inhibitors (ellipticine, teniposide, mAMSA) used as reference compounds. However, subtle structural modifications of the quinolones can strongly diminish the preferential genotoxicity in the prokaryotic test systems. PMID- 7476272 TI - Fragile site at Xq21 in Chinese hamster and its implications for the in vitro chromosomal aberration test. AB - Normal Chinese hamster primary fibroblasts were exposed to methotrexate, an antifolate agent known to induce fragile sites. In a total of 156 banded metaphases prepared from methotrexate-treated cells, 93 breaks and gaps were observed and mapped to the individual hamster chromosomes and their bands. About two-thirds (69%) of all breaks and gaps mapped to the band Xq21, which is significantly more than expected. Some other bands exhibited fragility but the effect was not significant. In 6% of cells, homozygous fragility of Xq21 was observed, suggesting that this is a common fragile site. It has been reported previously that various chemicals induce preferential localized fragility of Xq in Chinese hamster ovary (CHO) and V79 cell lines. Our results suggest that preferential fragility of Xq in the presence of chemical compounds represents examples of fragile-site induction. A tendency of the in vitro chromosomal aberration test to yield positive results in the absence of positive data in other test systems, such as the Ames test and the mouse bone marrow micronucleus test, might be due to fragile-site expression induced by chemical compounds under study. PMID- 7476273 TI - Biotransformation of genotoxic agents in marine sponges. Mechanisms and modulation. AB - Marine sponges do not appear to suffer from neoplastic diseases, in spite of possible high exposures resulting from their nature as sessile bottom filter feeders which pump large volumes of sea water. The assessment of several parameters related to the biotransformation of mutagens/carcinogens showed that the metabolic machinery of sponge medulla cells is mainly oriented towards detoxification, with some differences depending on species (Geodia cydonium or Tethya aurantium). Glutathione (GSH) levels were unexpectedly high in these cells, especially in Geodia, in which the concentration of this tripeptide was more than twice that measured in liver preparations from untreated rats, at least when related to the protein content. The oxidoreductive enzyme activities involved in the glutathione cycle were balanced in such a way as to favour a high GSH: oxidized glutathione (GSSG) ratio. GSH S-transferase activity was conversely rather low, compared to that of rat liver, and the dehydrogenases involved in the hexose monophosphate shunt were high in Tethya but low in Geodia. The metabolism of mutagens was investigated by using the Salmonella typhimurium his- strains TA100, TA98 and YG1024. Sponge S12 fractions failed to activate aflatoxin B1, benzo[a]pyrene and the two heterocyclic amines 3-amino-1-methyl-5H-pyrido[4,3 b]indole and 2-amino-3,4-dimethyl-imidazo[4,5-f]quinoline. Although far less efficiently than untreated rat liver S12 fractions, Geodia and especially Tethya preparations weakly activated the three aromatic amines 2-acetyl-aminofluorene, 2 aminofluorene and 2-aminoanthracene. On the other hand, sponge S12 fractions were remarkably efficient in decreasing the mutagenic potency of the direct-acting mutagens 4-nitroquinoline 1-oxide and sodium dichromate.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7476274 TI - Comparison of separated erythrocyte preparations and manual smears of bone marrow in showing micronucleus induction by clastogens and aneuploidogens in mouse. AB - The removal of nucleated cells from bone marrow cell suspensions by cellulose column separation and further purification in a Percoll gradient, coupled with slide preparation by a cytocentrifuge, produces uniform preparations of pure well spread erythrocytes. The use of separated erythrocytes has been a considerable improvement in the in vivo micronucleus (MN) assay, because the optimal cell morphology and the possibility of scoring a high number of polychromatic erythrocytes (PCEs) in a short time are factors contributing to the sensitivity of the assay. As the separation procedure may selectively retain cells containing MN, a study comparing the performance of the erythrocyte fractionation technique and standard manual smear preparation was conducted in male NMRI mice treated with a single injection of two clastogens (cyclophosphamide, 25 or 50 mg/kg, or methylmethane sulfonate, 40 or 80 mg/kg) or two aneuploidogens (colchicine, 0.5 or 1 mg/kg, or vincristine sulfate, 0.05 or 0.1 mg/kg). Both the preparation methods were clearly able to detect the MN-inducing and toxic effects of the treatments. After treatment with cyclophosphamide and methylmethane sulfonate, the frequency of micronucleated PCEs (MNPCEs) was, respectively, 1.6 and 1.8 times higher in the fractionated erythrocytes than in whole bone marrow. Morphological characterization of the MN indicated that this phenomenon was due to improved identification of small micronuclei in the flat cytospun cells of the fractionated erythrocyte preparations. On the other hand, the purified erythrocyte slides contained only about half the number of MNPCEs observed in standard smears, in samples collected from mice treated with colchicine.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7476275 TI - The effect of PHA stimulation on lymphocyte sub-populations in whole-blood cultures. AB - Although lymphocytes in phytohaemagglutinin (PHA)-stimulated whole blood cultures are routinely used to assess genotoxin-induced chromosome damage, very little information is available on the effect of PHA on the various cell populations present, and there appear to be no data for the protocols used in routine genotoxicity assays. In this study, we used flow cytometric analysis to examine the size/complexity of the white blood cell (WBC) population and the expression of key antigenic markers by the lymphocytes over a 96-h period following PHA stimulation. The changes in the WBC population are complex, and would seem to represent different populations dying out, remaining static or starting to divide. The initial decrease seen in overall cell numbers probably reflects death of the neutrophil and monocyte populations. The subsequent increase in cell numbers appears to be due to division of the lymphocytes and, by 96 h post stimulation, they comprise about half the total cell number and, as expected, > 90% are activated T-cells; it seems reasonable to assume that these represent the target cells in genotoxicity assays. Although we do not suggest that these findings should alter the routine conduct of clastogenicity assays using PHA stimulated whole-blood cultures, they indicate that such tests are empirical and that closer investigation will only confirm their relatively imprecise nature. PMID- 7476276 TI - Assessment of two protocols for the human lymphocyte cytokinesis-blocked micronucleus assay. AB - This study indicated that by adding cytochalasin B (6 micrograms/ml) at 24 h, the lymphocyte culture time for micronucleus (MN) assay could be shortened to 64 h. In both unirradiated and ex-vivo irradiated (2 Gy) lymphocytes from three populations, we found that the differences in MN yield obtained by our modified cytokinesis-blocked time frame and that recommended by Fenech and Morley (1985) were insignificant (P = 0.66-0.87). We believe that the shorter assay time may enhance the applicability of MN assay for the rapid assessment of ionizing radiation overexposures. PMID- 7476277 TI - Increased plasma levels of endothelin-1 in patients with hyperthyroidism. AB - Plasma concentrations of endothelin-1 (ET-1) were measured in 25 hyperthyroid subjects, 15 hypothyroid subjects, and 21 age-matched normal controls. In hyperthyroid patients, plasma concentrations of ET-1 were significantly higher than in the control group (P < .0001) and in hypothyroid patients (P < .0001). In contrast, no differences were found between hypothyroid patients and controls. Plasma levels of ET-1 were similarly elevated as in patients with Graves' disease and those with toxic adenoma. No correlations were found between plasma ET-1 levels, thyroid hormones, and thyrotropin (TSH) in hyperthyroid, hypothyroid, and euthyroid groups. The results of our study clearly indicate that in hyperthyroidism, circulating levels of ET-1 are strongly increased, although the pathogenesis of the increase is unclear. PMID- 7476278 TI - Chronic ethanol intake reduces the flux through liver branched-chain keto-acid dehydrogenase. AB - Chronic ethanol intake selectively increases concentrations of branched-chain amino acids (BCAA) in the liver. To determine whether a reduced oxidation plays a role in this effect, we measured substrate flux through branched-chain keto-acid (BCKA) dehydrogenase in livers of rats pair-fed liquid diets containing either 0% or 36% of total calories as ethanol for 21 days. Substrate (1.0 mmol/L ketoisocaproate [KIC]) fluxes in the liver of ethanol-fed and control rats were 225 +/- 18 and 319 +/- 27 mumol/h per whole liver (P < .05), respectively. We then studied whether this effect was due to either ethanol or the products of its metabolism, or to an alteration in the activity of BCKA dehydrogenase. Addition of ethanol (25 to 200 mmol/L) to the perfusion medium had no significant effect on the flux through BCKA dehydrogenase in the liver of control rats. Ethanol-fed rats had lower (P < .01) basal activity (0.84 +/- 0.11 v 1.39 +/- 0.12 U/g liver) and total activity (0.94 +/- 0.11 v 1.42 +/- 0.11 U/g liver) than control rats, but a similar activity state (90% +/- 4% v 99% +/- 4%) of BCKA dehydrogenase. In conclusion, chronic ethanol intake reduces the flux through liver BCKA dehydrogenase by decreasing the basal and total activity of BCKA dehydrogenase and not increasing the conversion of the enzyme to its inactive form. PMID- 7476279 TI - Effect of HCO3- on glutamine and glucose metabolism in lymphocytes. AB - Lymphocytes play a quantitatively important role in glutamine utilization in the body. We hypothesized that in metabolic acidosis characterized by decreased extracellular HCO3- concentration ([HCO3-]), glutamine utilization by lymphocytes may decrease to compensate partially for the increased uptake of glutamine by the kidneys for ammoniagenesis. This study was therefore designed to quantify the effect of extracellular [HCO3-] on glutamine metabolism in lymphocytes relative to glucose utilization. Mesenteric lymph node lymphocytes were incubated at 37 degrees C for 1 hour in Krebs-Henseleit buffer containing 0, 12.5, and 25 mmol/L HCO3- at a constant pH of 7.4 or 15.7 and 25 mmol/L HCO3- at a constant CO2 concentration of 1.25 mmol/L. Reducing extracellular [HCO3-] from 25 to 12.5 mmol/L at constant pH or from 25 to 15.7 mmol/L at constant CO2 concentration decreased glutamine utilization and the production of glutamate and ammonia. A reduction in [HCO3-] from 12.5 to 0 mmol/L further decreased glutamine utilization, as well as the production of all measured glutamine metabolites. Interestingly, decreasing [HCO3-] from 25 to 0 mmol/L had no significant effect on glucose metabolism, although the production of pyruvate (a minor product of glucose in lymphocytes) was decreased in the absence of medium HCO3-. The contribution of glutamine but not of glucose to lymphocyte adenosine triphosphate (ATP) production was decreased with reduced extracellular [HCO3-]. Thus, glucose was a more important fuel for lymphocytes than was glutamine at low [HCO3 ].(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7476280 TI - Comparison of serine and hippurate as precursor equivalents during infusion of [15N]glycine for measurement of fractional synthetic rates of apolipoprotein B of very-low-density lipoprotein. AB - Enrichment in hippurate has been measured to indicate precursor enrichment during glycine tracer infusion studies to estimate fractional synthetic rates of individual hepatic export proteins. However, hippurate tends to overestimate precursor enrichment. Since glycine is rapidly converted to serine by liver cells, we compared tracer enrichment in hippurate and serine with that of glycine incorporated into apolipoprotein (apo) B-100. Ten healthy control subjects were studied in the postabsorptive state during an 8-hour primed-constant infusion of [15N]glycine (10 mumol.kg-1.h-1). Apo B of very-low-density lipoprotein (VLDL) was isolated by standard ultracentrifugation and isopropanol precipitation. Glycine and serine were isolated from plasma and hydrolyzed apo B, hippurate was isolated from plasma, and [15N]enrichment was determined by gas chromatography mass spectrometry. Enrichment in serine and glycine isolated from apo B was identical at all time points, and their enrichment in apo B increased asymptotically, approaching an apparent plateau (mean +/- SD: 91% +/- 10% of calculated plateau at 8 hours) that was taken to represent hepatic protein precursor enrichment. Enrichment in both plasma serine and hippurate followed a biphasic pattern and continued to increase until the end of the study, raising the possibility that precursor enrichment had not reached a steady state during the study. The apo B plateau was lower (factor 0.76 +/- 0.27) than the final enrichment in hippurate and higher (factor 1.38 +/- 0.36) than that in plasma serine; however, predictions of protein precursor enrichment based on either metabolite were flawed by a large coefficient of variation (35% v 26%).(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7476281 TI - Changes in insulin sensitivity, glucose effectiveness, and B-cell function in regularly exercising subjects. AB - To determine the relative contributions of changes in glucose effectiveness, B cell function, and insulin sensitivity to changes in glucose tolerance upon exercise cessation in regularly exercising individuals, we studied seven young subjects who were performing aerobic exercise on a regular schedule. Each subject was studied 12 and 84 hours after the last bout of exercise with an intravenous glucose tolerance test (IVGTT) to quantify insulin sensitivity and glucose effectiveness at zero insulin (GEZI) using the minimal model of glucose kinetics. Additionally, B-cell function was quantified as the acute insulin response to glucose (AIRglucose), and intravenous glucose tolerance as the glucose disappearance constant (Kg). Twelve hours after the last bout of exercise, SI was 8.47 +/- 1.12 x 10(-5) min-1/pmol/L, as compared with 6.98 +/- 1.17 x 10(-5) min 1/pmol/L 84 hours after exercise (mean +/- SE, P = .005). No changes was observed in GEZI (0.020 +/- 0.004 min-1 at 12 hours v 0.019 +/- 0.002 min-1 at 84 hours, P = NS) or AIRglucose (588 +/- 213 pmol/L at 12 hours v 687 +/- 271 pmol/L at 84 hours, P = NS). Thus, the difference in intravenous glucose tolerance observed 12 hours after exercise as compared with 84 hours after the last bout of exercise (Kg, 2.91 +/- 0.70%/min at 12 hours v 2.23 +/- 0.60%/min at 84 hours, P < .05) would appear to be entirely related to a difference in SI and not to differences in glucose effectiveness or B-cell function. PMID- 7476282 TI - Low-carbohydrate diet alters intracellular glucose metabolism but not overall glucose disposal in exercise-trained subjects. AB - Dietary composition has been strongly implicated as an important determinant of in vivo insulin sensitivity. However, the metabolic alterations associated with extreme changes in diet have not been well described. We compared glucose metabolism after a standard diet ([STD] 35% fat, 51% carbohydrate, and 14% protein) with the effects of a 3-week adaptation to a low-carbohydrate, high-fat diet ([LCD] 75% fat, 8% carbohydrate, and 17% protein). Ten healthy men were studied using the euglycemic clamp technique, indirect calorimetry, and percutaneous vastus lateralis muscle biopsies for analysis of glycogen synthase (GS) and pyruvate dehydrogenase (PDH) activities in the basal and insulin stimulated states. Insulin-stimulated glucose disposal was unchanged (STD 46.1 +/ 4.3 v LCD 46.0 +/- 4.3 mumol/kg.min, P = NS), but marked alterations in the routes of glucose disposal were noted. Insulin-stimulated glucose oxidation (Gox) was markedly reduced following LCD (STD 18.6 +/- 1.9 v LCD 8.23 +/- 1.9 mumol/kg.min, P = .0001), and nonoxidative glucose metabolism (Gnox) was enhanced by LCD (STD 24.9 +/- 0.9 v LCD 38.9 +/- 4.3 mumol/kg.min, P = .03). Following LCD, both the total and active forms of PDH (PDHt and PDHa) were significantly depressed. After LCD, GS activates (FV0.1, %I, and A0.5) were unaffected in the basal state, but were greater than for STD (P = .004) after insulin stimulation. The apparent increase in the sensitivity of GS to activation by insulin following LCD correlated strongly with maximal O2 consumption ([VO2max] r = .97, P = .001), suggesting that physical conditioning interacted with the metabolic impact of LCD. In summary, LCD did not induce changes in net glucose disposal.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7476283 TI - Beef tallow diet decreases beta-adrenergic receptor binding and lipolytic activities in different adipose tissues of rat. AB - The effects of dietary fats consisting of different fatty acids on lipolytic activity and body fat accumulation were studied in rats. Sprague-Dawley male rats were meal-fed an isoenergetic diet based on either beef tallow or safflower oil for 8 weeks. Lipolytic activities in epididymal and subcutaneous adipose tissues were lower in the beef tallow diet group than in the safflower oil diet group. Body fat accumulation was greater in rats fed the beef tallow diet versus the safflower oil diet. Norepinephrine (NE) turnover rates used as an index of sympathetic activities in adipose tissues were lower in the beef tallow diet group. beta-Adrenergic receptor binding was determined with [3H]dihydroalprenolol. Binding affinities of beta-receptors in adipose tissues were significantly lower in the beef tallow diet group. Membrane fluidities of adipose tissues were also lower in the beef tallow diet group. Membrane fluidities were correlated with the affinities of the beta-receptor. We believe from these correlations that the decreases in beta-receptor binding affinities are due to the changes in membrane fluidities. The results of the present study suggest that intake of the beef tallow diet promotes body fat accumulation by reducing lipolytic activities resulting from lower beta-receptor binding and sympathetic activity in adipose tissues. PMID- 7476284 TI - Hyperinsulinemia in offspring of non-insulin-dependent diabetes mellitus patients: the role played by abnormal clearance of insulin. AB - Insulin resistance and hyperinsulinemia are often found in first-degree relatives of non-insulin-dependent diabetes mellitus (NIDDM) patients, and are currently considered a familial trait of this population at increased risk for diabetes. This study was undertaken to determine the role played by the metabolic clearance rate (MCR) of insulin (MCR-I) in the hyperinsulinism of these subjects. The proband population, consisting of 48 subjects aged 29.2 +/- 4.4 (mean +/- SD) years (18 men and 30 women; body mass index, 24.6 +/- 0.8 kg/m2; fasting plasma glucose, 4.54 +/- 0.37 mmol/L), was assigned in random order to four groups (I, II, III, and IV), each receiving a double insulin/glucose infusion (I, 0.025/2.0; II, 0.050/3.5; III, 0.100/6.0; and IV, 0.200/8.0 U/kg.h and mg/kg.min, respectively) to calculate MCR-I and MCR of glucose (MCR-G). Forty (14 men and 26 women) age- and body mass index-matched healthy individuals served as controls. All subjects had a normal response to an oral glucose tolerance test (75 g) according to World Health Organization criteria. Basal plasma insulin and C peptide levels in probands were significantly (P < .05) higher than in controls in each study group; similarly, MCR-I was significantly (at least P < .05) lower in probands than in controls in all groups. MCR-G was significantly (at least P < .05) decreased in probands as compared with controls of groups III and IV.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7476285 TI - Plasma levels of lipoproteins and apolipoproteins in congenital hypothyroidism: effects of L-thyroxine substitution therapy. AB - Thyroid status in humans is an important factor in the regulation of lipoprotein metabolism. There are several data on hypothyroidism in the adult population, but less information is available about congenital hypothyroidism. Since lipid metabolism at birth is substantially different from that of adults, it is not likely that the same abnormalities that occur in adult hypothyroidism are also present when this is diagnosed at early life. We studied 16 subjects with congenital hypothyroidism, seven at the time of diagnosis and also after normalization of thyroid hormone levels over a period of 2.0 +/- 1.0 months of substitution therapy with L-thyroxine (5.9 +/- 1.2 micrograms/kg/d) and nine already on L-thyroxine therapy for a period of 4.7 +/- 3.2 months. Thirty-nine apparently healthy subjects matched for age were selected as controls. In all subjects, total cholesterol (CHO), triglycerides (TG), low-density lipoprotein (LDL) cholesterol, high-density lipoprotein (HDL) cholesterol (HDL-C), apolipoproteins (apo) A-I and B, lipoprotein(a) [Lp(a)] thyrotropin (TSH), (LDL C), total and free thyroxine (T4), and triiodothyronine (T3) were determined. CHO, HDL-C, and apo A-I levels were significantly higher in patients at the time of diagnosis than in controls (respectively, P = .0079, .0007, and .0004), whereas TG, LDL-C, apo B, and Lp(a) levels were not significantly different. During L-thyroxine substitution therapy in these subjects, HDL-C and apo A-I levels significantly decreased (respectively, by a mean of -36.2% and -24.4%), with similar behavior in all subjects.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7476286 TI - Beta-adrenergic responsiveness of adenylate cyclase in human adipocyte plasma membranes in obesity and after massive weight reduction. AB - The aim of this study was to find out how beta-adrenergic responsiveness of adipocytes is altered in obesity and by weight loss and to investigate what mechanisms lead to potential alterations in responsiveness. Crude plasma membranes were prepared from adipocytes of massively obese and normal-weight individuals, as well as previously obese patients that had lost an average of 38% of their initial weight after bariatric surgery. Stimulation of adenylate cyclase by isoproterenol varied considerably in fat cell plasma membranes from different individuals. Crude fat cell plasma membranes from obese patients were less responsive to isoproterenol than those from normal-weight subjects, whereas those from postgastroplasty patients were hyperresponsive. The response was correlated negatively with cell size and positively with beta-adrenergic receptor density and with the ratio of beta-receptors and stimulatory G-proteins (Gs). There was no correlation with Gs content. However, differences in receptor density between small and large cells or normal-weight, obese, and post-bypass patients could not explain the observed differences in responsiveness to isoproterenol between the different groups. PMID- 7476287 TI - Lipoprotein(a) concentrations in non-insulin-dependent diabetes mellitus and borderline hyperglycemia: a population-based study. AB - The objective of the study was to compare lipoprotein(a) [Lp(a)] concentrations in population-based samples of individuals with non-insulin-dependent diabetes mellitus (NIDDM), borderline hyperglycemia, and normoglycemia. From 2,740 male Italian Telephone Company employees aged 40 to 59 years participating in a health screening, we selected all those with NIDDM (n = 100) plus a random sample of 950 nondiabetic individuals. Diabetes was defined as fasting plasma glucose (FPG) of at least 140 mg/dL or current use of hypoglycemic drugs. Among nondiabetic individuals, 854 were defined as normoglycemic (FPG < 115 mg/dL) and 95 were defined as borderline hyperglycemic (115 < FPG < 140 mg/dL). Lp(a) level was measured on frozen plasma by enzyme-linked immunosorbent assay. Lp(a) concentrations were similar in people with NIDDM, borderline hyperglycemia, and normoglycemia: 11.2 +/- 14, 14.1 +/- 20, and 13.9 +/- 18 mg/dL, respectively (F = 1.03). Accordingly, the proportion of subjects with Lp(a) levels of at least 30 mg/dL was comparable in the three groups (12%, 15%, and 14%; chi 2 = 3.95, P = .41). Results were not confounded by differences in age, body mass index (BMI), waist to hip ratio, plasma lipids, alcohol consumption, physical activity, and use of drugs. Furthermore, within the diabetic group Lp(a) levels were not significantly different for those on diet only versus those on oral agents (10.8 +/- 14.1 v 11.7 +/- 14.7, P = .7) or for people with FPG of at least 180 as compared with people with FPG less than 180 mg/dL (9.9 +/- 12.8 v 11.5 +/- 14.8, P = .5).(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7476288 TI - Defects in liver and muscle glycogen metabolism in neonatal and adult New Zealand obese mice. AB - Impaired glycogen synthesis is present in subjects at risk for developing non insulin-dependent diabetes mellitus (NIDDM), suggesting that it is a primary defect in NIDDM. To examine whether defects in glycogen metabolism are present at birth in an animal model of NIDDM, glycogen synthase (GS), glycogen phosphorylase (GP), and total glycogen content were measured in liver and quadriceps muscle of 1-day- and 20-week-old insulin-resistant New Zealand Obese (NZO) mice and control (NZC) mice. In livers of both neonatal and adult NZO mice, active GS was reduced by 54% and 36%, respectively, as compared with that in NZC mice (P < .03). Total liver GS activity was the same in neonates, but was 65% higher in adult NZO as compared with NZC mice (P < .02). Liver glycogen was 28% lower at birth in NZO mice (P < .03), but was 49% higher at 20 weeks of age. Active and total GP were the same in NZO and NZC animals, despite hyperinsulinemia in 20-week-old NZO mice. In muscle, active GS was reduced by 41% in both 1-day- and 20-week-old NZO mice (P < .02). Total GS was also lower in NZC mice at 1 day of age (P < .01), but not at 20 weeks. No differences were detected in GP activity or in total glycogen content in muscle. Therefore, reduced GS activity is an early defect present at birth in the insulin-resistant NZO mouse in both liver and muscle. However, it is not the sole determinant of the amount of glycogen deposited in tissues.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7476289 TI - Mental stress increases glucose uptake during hyperinsulinemia: associations with sympathetic and cardiovascular responsiveness. AB - Infusion of epinephrine and norepinephrine reduces insulin-mediated glucose disposal, ie, induces insulin resistance. Mental stress increases concentrations of both plasma catecholamines. However, the effect of acute mental stress on insulin-mediated glucose uptake has not been examined. We observed in pilot studies that a mental stress test (MST) during a euglycemic glucose clamp decreased blood glucose concentration. In a prospective study, euglycemic hyperinsulinemia was established during 120 minutes of glucose clamping; the subjects (N = 74) then underwent 5 minutes of intense mental arithmetics with infusion rates of glucose and insulin kept constant. During MST, plasma epinephrine and norepinephrine increased (by 0.23 +/- 0.02 and 0.50 +/- 0.05 nmol/L) together with blood pressure ([BP] by 18 +/- 8/9 +/- 1 mm Hg) and heart rate ([HR] by 21 +/- 1 beats per minute), with P less than .0001 for all changes. During mental stress, blood glucose concentration decreased by 0.4 +/- 0.1 mmol/L (P < .0001), followed by full recovery after another 10 minutes. Serum insulin was unchanged, indicating an acute but transient increase in glucose uptake. This finding was unrelated to age, sex, body mass, and BP status. Fifty-nine subjects with a decrease in glucose concentrations during MST were characterized by accentuated epinephrine response to MST (a change of 0.25 +/- 0.03 v 0.12 +/- 0.02 nmol/L, P = .001), increase in systolic BP (by 20 +/- 2 v 10 +/- 3 mm Hg, P = .008), and increase in HR (by 23 +/- 2 v 15 +/- 2 beats per minute, P = .008) as compared with 15 subjects with unchanged/increased glucose concentration.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7476290 TI - Erythrocyte insulin and insulin-like growth factor-I receptor tyrosine kinase activity in hypertension in pregnancy. AB - We have shown that preeclampsia is associated with insulin resistance. In the present study, we examined young normal, preeclamptic (PE), and gestational hypertensive (GH) nulliparous African-American women at term to investigate cellular determinants of this resistance and insulin and insulin-like growth factor-I (IGF-I) binding to partially purified erythrocyte receptors and receptor tyrosine kinase activity (TKA). Blood pressure was significantly elevated in PE and GH subjects as compared with controls. Insulin binding was similar in number and affinity in the three groups (femtomoles per microgram). IGF-I binding was increased in PE subjects as compared with either normals or GH subjects (0.2 +/- 0.02, 0.15 +/- 0.01, and 0.14 +/- 0.02 fmol/microgram protein). Insulin receptor TKA was increased in PE subjects as compared with normals when assessed either per microgram protein or per femtomole insulin binding (P < .01). In contrast, IGF-I-potentiated TKA was elevated in PE subjects only when assessed per microgram protein (P < .03). Thus, the increased number of IGF-I receptors in erythrocytes of PE subjects yields a net increase in receptor tyrosine kinase. Also, there is an augmentation of insulin receptor TKA in PE subjects. Together, these two alterations may be a compensatory mechanism for the insulin resistance associated with hypertensive diseases of pregnancy. PMID- 7476291 TI - Chromium improves insulin response to glucose in rats. AB - The effects of chromium (Cr) supplementation on insulin secretion and glucose clearance (KG) during intravenous glucose tolerance tests (IVGTTS) were assessed in rats with impaired glucose tolerance due to dietary Cr deficiency. Male Wistar rats were maintained after weaning on a basal low-Cr diet containing 55% sucrose, 15% lard, 25% casein. American Institute of Nutrition (AIN)-recommended levels of vitamins, no added Cr, and an altered mineral content as required to produce Cr deficiency and impaired glucose tolerance. The Cr-supplemented group ([+Cr] n = 6) were provided with 5 ppm Cr as CrCl3 in the drinking water, and the Cr deficient group ([-Cr]n = 5) received purified drinking water. At 12 weeks on the diet, both groups of rats were hyperinsulinemic (+Cr, 103 +/- 13; -Cr, 59 +/- 12 microU/mL) and normoglycemic (+Cr, 127 +/- 7; -Cr, 130 +/- 4 mg/dL), indicating insulin resistance. After 24 weeks, insulin levels were normal (+Cr, 19 +/- 5; Cr, 21 +/- 3 microU/mL) and all rats remained normoglycemic (+Cr, 124 +/- 8; -Cr, 131 +/- 6 mg/dL). KG values during IVGTTS were lower in -Cr rats (KG = 3.58%/min) than in +Cr rats (KG = 5.29%/min), correlating with significantly greater 40 minute glucose areas in the -Cr group (P < .01). Comparisons of 40-minute insulin areas indicated marked insulin hyperresponsiveness in the -Cr group, with insulin secretory responses increased nearly twofold in -Cr animals (P < .05). Chromium deficiency also led to significant decreases in cyclic adenosine monophosphate (cAMP)-dependent phosphodiesterase (PDE) activity in spleen and testis (P < .01). In these studies, Cr deficiency was characterized by both beta-cell hypersecretion of insulin and tissue insulin resistance that were associated with decreased tissue levels of cAMP PDE activity. PMID- 7476292 TI - Endothelin-1 decreases glucose, inhibits glucagon, and stimulates insulin release in the rat. AB - The effects of endothelin-1 (ET-1) infusion at 0, 25, 50, and 75 ng/kg/min on blood glucose, insulin, and ET-1 levels were determined in anesthetized rats. In a separate group of rats, ET-1 was infused at 75 ng/kg/min and glucagon and glucose levels were determined. In another group of rats, the effect on blood glucose of glucagon infusion at 0.2 ng/kg/min with ET-1 infusion at 75 ng/kg/min for 30 minutes was determined. Glucose decreased 10 minutes after initiation of ET-1 infusion at 75 ng/kg/min and at 15 minutes during ET-1 infusion at 25 and 50 ng/kg/min. After 45 minutes, glucose decreased by 1.05 +/- 0.1, 1.44 +/- 0.11, and 1.39 +/- 0.22 mmol/L and ET-1 increased by 4.4 +/- 0.8, 5.2 +/- 1.2, and 11.2 +/- 0.8 pmol/L during ET-1 infusion at 25, 50, and 75 ng/kg/min, respectively. Insulin levels increased during ET-1 infusion of 50 ng/kg/min at 30 and 45 minutes by 300 +/- 75 and 405 +/- 120 pmol/L, respectively. During ET-1 infusion of 75 ng/kg/min, insulin increased at 45 minutes by 570 +/- 180 pmol/L. Glucagon decreased during ET-1 infusion at 15 minutes associated with a decrease in glucose. Glucagon levels subsequently returned to baseline values despite a continued decline in glucose levels. Glucagon infusion at 0.2 microgram/kg/min prevented the early ET-1-induced hypoglycemia. These findings demonstrate that ET 1 decreased blood glucose initially associated with a decrease in glucagon and subsequently associated with enhanced insulin release. PMID- 7476293 TI - Gender differences in the development of hyperlipemia and atherosclerosis in hybrid hamsters. AB - In response to a diet enriched in saturated fat and cholesterol (CH), male Syrian hamsters develop hyperlipemia and changes of early atherosclerosis. However, it has not been determined if female hamsters are equally susceptible to an atherogenic diet. Male and female hamsters of the F1B hybrid strain (Bio Breeders, Fitchburg, MA) were fed either a chow diet or this diet (HiFat) with added saturated fat (10% coconut oil) and CH (0.05%) for up to 12 weeks. Female hamsters ate significantly more than males, and with the HiFat diet gained threefold more weight than males. However, with the HiFat diet, serum triglycerides (TGs) and CH were markedly increased only in male hamsters. Furthermore, only in males was there a significant increase in stainable fat in the aorta that corresponded to an increase in subintimal foam cells. In freely feeding males, the largest percentage increase in serum CH was in the TG-rich fraction of lipoproteins. After females were castrated, serum TG and CH levels increased to the same extent as in males. These studies demonstrate a profound gender difference in response to an atherogenic diet in these hamsters that has parallels to the lipid patterns of humans and their susceptibility to atherosclerosis. PMID- 7476294 TI - Metabolic and hormonal responses to adrenoceptor antagonists in 48-hour-starved exercising rats. AB - The influence of 48 hours of starvation on sympathoadrenal regulation of nutrient utilization was investigated in rats. To assess the role of alpha- and beta adrenoceptors, rats were studied during alpha- and beta-blockade. Energy metabolism was measured using indirect calorimetry before, during, and after moderate swimming exercise (approximately 60% maximal O2 consumption [VO2max]). Additionally, blood samples were taken for determination of nutrient and hormone concentrations. In 48-hour-starved rats, under baseline conditions, there was a reduction in energy expenditure (EE) accompanied by a shift toward fat oxidation (fat-ox) in comparison to fed rats. Exercise-induced responses in EE, fat-ox, and carbohydrate oxidation (CHO-ox) did not differ from those in fed rats. In starved rats, a stronger response to exercise of the sympathoadrenal system was observed. In comparison to control 48-hour-starved rats, blockade of alpha- and beta adrenoceptors led to a reduction in the exercise-induced increase in EE and fat ox. The rate of CHO-ox was slightly reduced after blockade of either adrenoceptor type. Alpha-blockade prevented the exercise-induced increase in blood glucose. Plasma free fatty acid (FFA) was not affected. Blood lactate, plasma insulin, norepinephrine (NOR), and epinephrine (EPI) were increased after alpha-blockade. Due to beta-blockade, exercise-induced increases in glucose and FFA were prevented. Blood glucose even declined below the baseline value. EPI showed an exaggerated increase, and NOR showed a smaller increase. Results obtained in starved rats support the idea that alpha-adrenoceptor blockade-induced changes in energy metabolism are the result of a diminished oxygen supply due to diminished circulation.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7476295 TI - Negative impact of cancer chemotherapy on protein metabolism in healthy and tumor bearing rats. AB - Although chemotherapeutic agents are widely used in the treatment of cancer, few experimental data are available on their effects on host N metabolism. We studied the effects of a single intraperitoneal (IP) injection of cyclophosphamide ([CYP] 120 mg/kg), 5-fluorouracil ([5-FU], 50 mg/kg), cisplatinum ([CDDP], 5 mg/kg), or methotrexate ([MTX], 30 mg/kg). N balance was studied for 6 days following chemotherapy in healthy rats (n = 40) and in rats bearing Morris Hepatoma 7777 ([MH7777] n = 40) in a situation comparable to that of human cancer (tumor burden < 0.2% of body weight, moderate anorexia, and weight loss). In healthy rats, all drugs induced transient body weight loss, anorexia, and poor N balance. At day 6 posttreatment, all animals had resumed normal feed intake and positive N balance except CDDP-treated rats, which showed continued weight loss and poor N balance. CDDP and MTX exhibited antitumor activity; however, CDDP induced diarrhea in six of eight tumor-bearing rats. Drug-induced anorexia was more severe in tumor bearing than in healthy treated rats. N balance was more severely decreased in MH7777-bearing rats than in healthy treated animals in response to 5-FU (159 +/- 36 v 273 +/- 27 mg N/2 d) and MTX (-66 +/- 36 v 153 +/- 37 mg N/2 d) at days 3 to 4 postinjection. These results establish the presence of drug-specific effects on host N balance and the existence of a drug-tumor interaction for N metabolism in the tumor-bearing host. PMID- 7476296 TI - Bromocriptine inhibits in vivo free fatty acid oxidation and hepatic glucose output in seasonally obese hamsters (Mesocricetus auratus). AB - Seasonally obese hyperinsulinemic hamsters were treated for 5 weeks with bromocriptine (500 to 600 micrograms per animal) and tested for drug effects on energy balance, body fat stores, nocturnal whole-body free fatty acid (FFA) metabolism and hepatic glucose output, and diurnal glucose tolerance. After 5 weeks, bromocriptine treatment reduced retroperitoneal fat pad weight by 45% without altering either daily food consumption or end-treatment total daily energy expenditure. Also, 5 weeks of treatment improved the diurnal glucose tolerance, resulting in a 47% and 33% decrease in the area under glucose and insulin curves, respectively. After 4 weeks, bromocriptine treatment reduced nocturnal lipolysis by 28%, palmitate rate of appearance into plasma by 30%, palmitate oxidation by 33%, and hepatic glucose output by 28%. Moreover, these reductions were accompanied by a 75% reduction in plasma insulin concentration. The data suggest that bromocriptine may improve diurnal glucose tolerance in part by inhibiting the preceding nocturnal lipolysis and FFA oxidation. Reductions in nocturnal FFA oxidation and hepatic glucose production may result from bromocriptine's influences on circadian organization of hypothalamic centers known to regulate these activities. Available evidence suggests that bromocriptine may impact this neuroendocrine organization of metabolism by increasing the dopamine to noradrenaline activity ratio in central (hypothalamic) and peripheral (eg, liver and adipose) target tissues. PMID- 7476297 TI - Ethanol induces a paradoxical simultaneous increase in circulating concentrations of insulin-like growth factor binding protein-1 and insulin. AB - The aim of this study was to examine the effect of acute alcohol intake on circulating concentrations of insulin, C-peptide, insulin-like growth factor (IGF) binding protein-1 (IGFBP-1), and plasma glucose levels. We measured these parameters for 12 hours after administration of 0, 0.5, or 1.0 g ethanol/kg body weight to nine healthy volunteers between 7:00 and 7:45 PM according to a randomized, double-blind, crossover design. Following a snack at 9:00 PM, plasma insulin (P < .05) and C-peptide (P < .01) concentrations were significantly increased at 10:00 PM in the 1.0-g group as compared with the control group. C peptide to insulin molar ratios were significantly higher (P < .05) in both ethanol groups at 10:00 PM and 2:00 AM than in the control group. No differences were observed in plasma glucose levels between the three groups. Plasma IGFBP-1 levels showed a dose-dependent increase in the ethanol groups, and remained increased from 10:00 PM for 3 hours (P < .05 or less) at the lower dose and for 6 hours (P < .05 or less) at the higher dose. These observations indicate that ethanol-induced postprandial hyperinsulinemia is due to increased insulin secretion and that alcohol may increase hepatic insulin extraction. The lack of any effect on plasma glucose levels suggests that alcohol intake must be associated with decreased insulin sensitivity. Alcohol intake results in a paradoxical increase in peripheral concentrations of IGFBP-1 despite simultaneous hyperinsulinemia. This implies that ethanol has a direct stimulatory effect on hepatic IGFBP-1 synthesis. PMID- 7476298 TI - Beta-cell function and replication in spontaneously hypertensive rats. AB - We examined beta-cell function and replication in spontaneously hypertensive rats (SHR) and age-matched Wistar-Kyoto rats (WKY). Rats were subjected to 90% pancreatectomy (Px) or sham operation at the age of 8 weeks, and islet function and regeneration were examined 4 weeks after surgery. Plasma glucose levels were higher in SHR than in WKY (509 +/- 38 v 325 +/- 109 mg/dL, P < .0001) 1 week after Px and throughout the experimental period. Plasma glucose responses to intravenous injection of glucose (0.5 g/kg body weight) were not different in the sham-operated animals of the two strains, whereas plasma insulin responses were greater in SHR than in WKY. No insulin responses to glucose were observed in either strain of Px rats. The insulin content of the remnant equivalent (6.7 +/- 2.1 v 4.2 +/- 0.4 micrograms, P < .05) and whole pancreas (156.7 +/- 10.7 v 123.8 +/- 23.5 micrograms, P < .01) in sham-operated rats was greater in SHR than in WKY. However, insulin content was lower (P < .05) in Px-SHR (1.0 +/- 0.2 microgram) than in Px-WKY (3.9 +/- 1.7 micrograms). Histological examination showed that fibrotic degeneration of islets was much greater in Px-SHR than in Px WKY. These data strongly suggest that the beta cells of SHR were more vulnerable to reduction of islet mass than those of WKY. Our data also suggest that hyperinsulinemia and/or insulin resistance in SHR has a deleterious effect on beta-cell replication. PMID- 7476299 TI - Fish oil feeding improves muscle glucose uptake in tumor necrosis factor-treated rats. AB - This study was conducted to characterize the effects of fish oil and sunflower oil on hepatic glucose production and peripheral glucose utilization during infusion of saline or tumor necrosis factor (TNF), using the euglycemic hyperinsulinemic clamp technique combined with a primed-constant tracer infusion of high-performance liquid chromatography-purified 3H-3-glucose for estimation of whole-body glucose appearance and utilization rates. Insulin 10 mU/kg.min was infused to reach a plasma insulin level of 200 microU/mL. 14C-1-deoxyglucose (14C DG) uptake was also measured in specific tissues following intravenous bolus administration. The results showed that during a hyperinsulinemic-euglycemic clamp, infusion of TNF 20 micrograms/kg for 3 hours resulted in a significant reduction of glucose infusion and a significant increase of hepatic glucose production in both dietary groups as compared with saline infusion, indicating a state of insulin resistance induced by TNF. The results also showed that TNF infusion significantly decreased the rate of 14C-DG uptake in muscle in the sunflower oil group but not in the fish oil group, suggesting that fish oil is able to restore to normal the glucose utilization impaired by TNF. These observations suggest that in hyperinsulinemic and euglycemic conditions, prefeeding with fish oil significantly improves glucose uptake in muscle tissue, but does not alter the increase in hepatic glucose production during TNF infusion. PMID- 7476300 TI - Are specific serum insulin levels low in impaired glucose tolerance and type II diabetes?: measurement with a radioimmunoassay blind to proinsulin, in the population of Wadena, Minnesota. AB - It has been suggested that serum insulin levels in subjects with recently diagnosed type II diabetes have been overestimated, and that after correction for proinsulin, true insulin levels are depressed rather than elevated. We tested this possibility in a cross-sectional study of a population-based sample of 328 adults living in Wadena, a Minnesota community in which residents are of northern European background. Specificity of insulin measurements was provided by an antibody blind to proinsulin and its major metabolite. Oral glucose tolerance and liquid mixed-meal (Ensure-Plus) tests were performed on separate days. Mean insulin levels before and 90 minutes after the mixed meal were as follows. Among 302 randomly ascertained adults not previously known to have diabetes, both fasting and postmeal levels in subjects with impaired glucose tolerance (IGT) and newly identified type II diabetes were equal to or greater than levels in subjects with normal glucose tolerance (fasting: normal 52 pmol/L, IGT 78, new type II 87; postmeal: 317, 565, and 406, respectively). The fasting insulin to glucose ratio was significantly increased in IGT and new type II diabetes subjects. Among 26 established (previously known) type II diabetic subjects not taking insulin, fasting levels were elevated and postmeal levels were normal in absolute terms (75 and 328), but were normal or low with respect to plasma glucose. Relationships among the groups were not materially changed by adjustment for body mass index (BMI), sex, age, or blood pressure. There was marked overlap of individual insulin levels from group to group. In summary, randomly selected adults in Wadena with IGT or asymptomatic diabetes showed, on average, elevated insulin levels, but physician-diagnosed diabetes was associated with relative diminution of serum insulin. In this population, the current view of insulin resistance in "early" diabetes was supported by insulin-specific measurements. PMID- 7476301 TI - Fuel metabolism in growth hormone-deficient adults. AB - Apart from being a stimulator of longitudinal growth, growth hormone (GH) regulates fuel metabolism in children and adults. A halfmark is mobilization of lipids, which involves an inhibition of lipoprotein lipase activity in adipose tissue and activation of the hormone sensitive lipase. Suppression of basal glucose oxidation and resistance to insulin are other important effects. This may cause concern during GH substitution in GH-deficient adults, some of whom may present with insulin resistance due to concomitant abdominal obesity. However, there are data to suggest that the GH-induced reduction in fat mass and increase in lean body mass may offset the insulin antagonistic actions of the hormone. The nitrogen-retaining effects of GH seem to involve a direct stimulation of protein synthesis in addition to secondary effects such as generation of insulin-like growth factor-I (IGF-I), hyperinsulinemia, and promotion of lipolysis. Thus, during periods of substrate affluence, GH acts in concert with insulin and IGF-I to promote protein anabolism. Postabsorptively, GH is primarily lipolytic and thereby indirectly protein-sparing. This effect becomes further accentuated with more prolonged fasting. In that sense, GH is unique by its preservation of protein during both feast and famine. These fuel metabolic effects add merit to the principle of GH substitution in hypopituitary adults. PMID- 7476302 TI - Metabolic effects of insulin-like growth factor-I: a focus on insulin sensitivity. PMID- 7476303 TI - Carbohydrate metabolism in primary growth hormone resistance (Laron syndrome) before and during insulin-like growth factor-I treatment. AB - Among 43 patients with Laron syndrome followed in our clinic, we were able to study the carbohydrate metabolism from infancy into adult age in 30 patients. During infancy, fasting blood glucose levels were in the hypoglycemic range (mean +/- SD, 3.5 +/- 1.2 mmol/L) and increased at the end of a delayed puberty to 4.6 +/- 0.6 mmol/L. Fasting plasma insulin was higher than expected for concomitant glucose levels, and several of the 20 patients who underwent an oral glucose tolerance test (OGTT) had glucose intolerance and relatively high insulin levels. In adult patients, insulinopenia developed and one 38-year-old patient developed non-insulin-dependent diabetes mellitus (NIDDM) with subsequent need for insulin therapy. Continuous insulin-like growth factor-I (IGF-I) treatment of a pubertal patient with glucose intolerance and hyperinsulinemia normalized both responses. In conclusion, long-term IGF-I deficiency leads to insulin resistance, which is reversed by exogenous IGF-I administration. PMID- 7476304 TI - Insulin-like growth factors (IGFs) and IGF-I treatment in the adolescent with insulin-dependent diabetes mellitus. AB - Insulin-dependent diabetes mellitus (IDDM) during adolescence is associated with complex derangements of the growth hormone (GH)/insulin-like growth factor (IGF) axis. Despite GH hypersecretion, IGF-I levels and IGF bioactivity are reduced. The diabetogenic effects of GH are well established, and GH hypersecretion has been implicated in the deterioration in glycemic control during adolescence and in the development of microangiopathy. Insulin deficiency or reduced portal delivery of insulin plays a central role in the development of these abnormalities, and although continuous subcutaneous insulin delivery may improve plasma IGF-I levels, it does not necessarily suppress GH levels. Recombinant IGF I has been proposed as an adjunct to conventional insulin therapy, as restoring circulating IGF-I levels might lead to GH suppression. Placebo-controlled studies have shown a consistent reduction in GH secretion and related improvements in insulin sensitivity following a single subcutaneous IGF-I injection (40 micrograms/kg). Repeated daily subcutaneous IGF-I administration for 1 month resulted in a sustained increase in IGF-I levels, as well as a reduction in GH secretion and insulin requirements. There was no increase in hypoglycemia or other adverse effects. Recombinant IGF-I used in conjunction with insulin may therefore provide an additional approach to the management of IDDM during adolescence, allowing correction of abnormalities in the GH/IGF axis and leading to improved control and, hence, reduced risk of long-term complications. However, this hypothesis needs to be rigorously tested in long-term placebo-controlled studies. PMID- 7476305 TI - Insulin-like growth factor binding proteins as glucoregulators. AB - Circulating insulin-like growth factors (IGFs) represent an important pool of potential hypoglycemic activity, which is largely inhibited by their sequestration in a heterotrimeric complex comprising growth factor, IGF-binding protein-3 (IGFBP-3), and acid-labile subunit (ALS). Less than 1% of total IGFs circulate in the free form, yet even this amount might contribute significantly to circulating insulin-like activity. The ternary binding protein complex appears to inhibit insulin-like activity of bound IGFs by preventing their egress from the circulation. Although the integrity of this complex might be affected by limited proteolysis of IGFBP-3 in pregnancy and catabolic conditions, the evidence that this increases IGF bioavailability, and thus hypoglycemic potential, is as yet unclear. However, in patients with IGF-II-secreting tumors, hypoglycemia may result from a failure of the ternary complex to adequately sequester the IGFs. Improvement in complex formation, by treatment with corticosteroids or growth hormone, alleviates the hypoglycemia, even if (as seen with growth hormone treatment) IGF-II hypersecretion persists. In these patients, blood glucose levels are inversely correlated with IGFBP-2 levels, suggesting that this protein might play a part in transporting IGFs to their target tissues. Conversely, ALS levels correlate positively with blood glucose, emphasizing the importance of the ternary complex in preventing hypoglycemia. Unlike the other IGF-binding proteins, IGFBP-1 is acutely regulated in the circulation, in a manner consistent with its acting as a glucose counterregulator. It might act in this way by inhibiting the activity of free IGFs in the circulation.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7476306 TI - Effected of insulin and insulin-like growth factor-I on glucose transport and its transporters in soleus muscle of lean and obese mice. AB - The mechanisms underlying insulin and insulin-like growth factor-I (IGF-I) action on glucose transport share similar processes leading to Glut 4 translocation after respective receptor activation. Among these steps are phosphorylation of insulin receptor substrate-1 (IRS-1) and activation of phosphatidylinositol-3 kinase (P13-kinase). This enzyme could be involved in stimulated glucose transport in muscle, since its inhibitor, wortmannin, blocks the hormonal effect in muscle. P13-kinase is activated by insulin and IGF-I in a rapid and transient manner in incubated soleus muscles. When P13-kinase activation was studied in muscle of obese insulin-resistant mice, there was a marked alteration in the response to insulin both in vivo and in vitro. P13-kinase activation by IGF-I was also altered in obese mice, although to a lesser degree. PMID- 7476307 TI - Role of the time factor in signaling specificity: application to mitogenic and metabolic signaling by the insulin and insulin-like growth factor-I receptor tyrosine kinases. AB - The signal transduction pathways activated by hormones, growth factors, and cytokines show an extraordinary degree of cross-talk and redundancy. This review addresses the question of how the specificity conferred at the binding step is maintained through the signaling network despite the convergence of multiple signals on common efferent pathways such as mitogen-activated protein (MAP) kinase. The mechanism of receptor activation by ligand-induced dimerization provides a signaling device with both a switch and a timer. The role of the time factor, ie, of signaling kinetics, as a determinant of selectivity is discussed with emphasis on the receptor tyrosine kinases and cytokine receptors, and especially mitogenic versus metabolic signaling by insulin and insulin-like growth factor-I (IGF-I). PMID- 7476308 TI - Intracellular signaling by growth factors. AB - Growth factors are involved in a variety of cellular responses such as growth, differentiation, migration, metabolism, and transformation. Binding of the growth factor to its corresponding cell surface receptor results in activation of the receptor's intrinsic tyrosine kinase activity, and subsequently in activation of complex multistep signal transduction cascades. Activation of these interconnected signaling pathways eventually leads to a biological response, which involves changes in gene expression and protein synthesis. The biological response has been shown to be receptor-specific and also cell-type (tissue) specific, indicating that various receptors activate distinct signal transduction pathways in one tissue and that one receptor activates different pathways in various tissues. What determines receptor specificity and tissue specificity? In this context, this article will focus on certain receptors with intrinsic tyrosine kinase activity, including receptors for platelet-derived growth factor (PDGF), epidermal growth factor (EGF), insulin, and nerve growth factor (NGF). PMID- 7476309 TI - Metabolic effects of growth hormone in humans. AB - Growth hormone (GH) has acute actions to stimulate lipolysis and ketogenesis after 2 to 3 hours, effects that may be important in the adaptation to stress and fasting. This is accompanied by a decrease in insulin sensitivity in both liver and muscle. These combined effects may be very deleterious to insulin-dependent diabetic patients, in whom increased GH secretion may precipitate and maintain acute metabolic derangement (ketoacidosis) and be a major initiator of the dawn phenomenon. On the other hand, augmented GH secretion plays a beneficial role in the defense against hypoglycemia, in particular during prolonged hypoglycemia and in patients with impaired ability to secrete other counterregulatory hormones appropriately. It is also certain that GH is a potent anabolic hormone in terms of promoted nitrogen retention, but the extent to which these well-known actions are direct or secondary to hyperinsulinemia, increased activity of insulin-like growth factors (IGFs), or release of protein-conserving lipid intermediates has eluded precise characterization. PMID- 7476310 TI - Free insulin-like growth factors in human obesity. AB - It is well established that spontaneous and stimulated growth hormone (GH) secretion is diminished in human obesity. In contrast to classic GH deficiency, obesity is not associated with hypopituitary levels of circulating total (extractable) insulin-like growth factor-I (IGF-I) and reduced somatic growth. Thus, the riddle of "normal growth without GH" in obese children and the mechanisms behind the GH suppression have remained unsolved. Insulin reduces hepatic production of IGF-binding protein-1 (IGFBP-1), an in vitro inhibitor of IGF bioactivity, and it has been suggested that the obesity-related hyperinsulinemia may increase free (bioactive) IGF in vivo by reducing the concentration of IGFBP-1. We have recently developed a method that during near in vivo conditions isolates the free, unbound fractions of IGF-I and IGF-II in human serum. Using this method, we have determined overnight fasting serum levels of free IGFs in obese subjects and compared the results with levels of total (extractable) IGFs, IGFBPs, GH, and insulin. The study included 92 healthy subjects (56 males and 36 females) allocated to three age-matched groups depending on body mass index (BMI): 31 controls (BMI < or = 25), 33 subjects with moderate obesity (25 < BMI < 30), and 28 subjects with severe obesity (BMI > or = 30). Fasting serum insulin correlated positively (r = .61, P < .0001) with BMI and was significantly elevated in moderate and severe obesity (P < .05).(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7476311 TI - The role of growth hormone/insulin-like growth factors in adipocyte differentiation. AB - Growth of the adipose tissue results from both the enlargement of mature adipocytes and the formation of new adipocytes from adipocyte precursor cells. The differentiation process of adipocyte precursor cells is controlled by a variety of hormones. Clinical observations indicate that growth hormone (GH) and insulin-like growth factor I (IGF-I) are able to influence the growth of the adipose organ. Recent in vitro studies using cultures of clonal and primary adipocyte precursor cells have elucidated the role of GH and IGF-I in adipocyte differentiation. From the studies it can be concluded that GH is able to enlarge the pool of adipocyte precursor cells capable of differentiating into mature adipocytes, which occurs under the control of other adipogenic hormones. However, due to its metabolic action, GH is also able to reduce the volume of mature adipocytes and thus the net result of its biological action is aimed at reducing body fat. IGF-I stimulates the differentiation process by inducing critical cell divisions of adipocyte precursor cells necessary for their differentiation. IGF I, which is known to be regulated by GH and several nutritional factors, may exert its effects in the adipose tissue in an autocrine/paracrine and endocrine way. This review summarizes the results of recent studies investigating the role of GH and IGF-I in adipocyte differentiation. PMID- 7476312 TI - Nutritional regulation of insulin-like growth factor-I. AB - Several lines of evidence indicate that in the human, insulin-like growth factor I (IGF-I) is nutritionally regulated. Both energy and protein availability are required for maintenance of IGF-I. Measurements of serum IGF-I constitute a sensitive means for monitoring the response of acutely ill patients to nutritional intervention. Serum IGF-I may also serve as a marker for evaluation of nutritional status. Our findings and those of others in animal models suggest that nutrients influence synthesis and action of IGF-I and its binding proteins (IGFBPs) at multiple levels. In fasting, liver growth hormone (GH) binding is decreased, providing one explanation for decreased IGF-I. In protein restriction, GH receptors are maintained, but there is evidence for a postreceptor defects. The latter results from pretranslational and translational defects. Amino acid availability to the hepatocytes is essential for IGF-I gene expression. Protein malnutrition not only decreases IGF-I production rate, but also enhances its serum clearance and degradation. Finally, there is evidence for selective organ resistance to the growth-promoting effects of IGF-I in protein-restricted rats. PMID- 7476313 TI - The insulin-like growth factor system in vascular smooth muscle: interaction with insulin and growth factors. AB - Vascular smooth muscle cells (SMCs) occur throughout the vascular tree and have important physiological functions. They are also involved in pathological processes such as development and progression of atherosclerotic lesions, restenosis following angioplasty, and in hypertension. This review is focused on the role of the insulin-like growth factor (IGF) system in proliferation, migration, and hypertrophy of vascular SMCs and its interaction with insulin and other growth factors. The IGF-I receptor is highly expressed in SMCs in intact arteries and in cultured SMCs and is activated by binding of IGF-I to the two alpha-subunits. Insulin and IGF-II from the circulation can interact with the IGF I receptor at higher concentrations. Insulin receptors are few or absent in SMCs and circulating insulin concentrations in vivo are probably too low for a direct action of insulin on the IGF-I receptor in SMCs. Receptor activation initiates a number of signal transduction pathways. Increased phosphatidylinositol turnover and calcium mobilization correlates with actin filament reorganization and stimulation of directed migration of the SMC in a gradient of IGF-I. The effects of IGF-I receptor activation on signal transduction pathways (eg, the MAP kinase cascade) implicated in DNA synthesis and proliferation are weak and this correlates with the meager mitogenic activity of IGF-I in SMC. Several components of the IGF-system in SMC are regulated by growth factors such as platelet-derived growth factor (PDGF)-BB and basic fibroblast growth factor (bFGF).(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7476314 TI - The role of growth hormone, insulin-like growth factors (IGFs), and IGF-binding proteins in experimental diabetic kidney disease. AB - Early renal changes in type I diabetes are characterized by an increase in renal size, glomerular volume, and kidney function, and later by development of mesangial proliferation, accumulation of glomerular extracellular matrix, and increased urinary albumin excretion (UAE). Growth hormone (GH) and insulin-like growth factors (IGFs) have a long and distinguished history in diabetes mellitus, with possible participation in the development of long-term complications. In experimental diabetes in dwarf rats with isolated GH and IGF-I deficiency, a slower and lesser renal and glomerular hypertrophy is observed as compared with diabetic control animals with intact pituitary. Furthermore, diabetic dwarf rats with a diabetes duration of 6 months display a smaller increase in UAE, indicating that GH and IGF-I may be involved in the development of diabetic kidney changes. In line with this, administration of octreotide to streptozotocin (STZ)-diabetic animals with normal pituitary inhibits initial renal growth without affecting blood glucose levels, and 6 months' administration of octreotide to diabetic rats reduces long-term renal/glomerular hypertrophy and UAE. In addition, the initial increase in renal size and function in experimental diabetes is preceded by an increase in renal IGF-I, IGF-binding proteins (IGFBPs), and IGF-II/mannose-6-phosphate receptor (IGF-II/Man-6-P receptor) concentration. Finally, specific changes occur in renal GH-binding protein (GHBP) mRNA, IGF-I receptor mRNA, and IGFBP mRNA expression in long-term diabetes. In conclusion, the knowledge we have today indicates that GH and IGFs, through a complex system consisting of GHBP, IGFs, IGF receptors, and IGFBPs, may be responsible for both early and late renal changes in experimental diabetes. PMID- 7476315 TI - The role of growth factors in the development of diabetic retinopathy. AB - Proliferation of retinal blood vessels is one of the most striking features of advanced diabetic retinopathy. This feature has led to the conclusion that the normal balance of growth factors, which usually serves to keep angiogenesis in check, is disturbed in diabetic retinopathy. A considerable amount of work has been performed in the field of angiogenesis within the last decade. Much of this is applicable to diabetic eye disease, but due to the lack of an animal model, few studies have been performed directly on models of diabetic retinopathy. This review examines the literature as it relates to diabetic retinopathy. PMID- 7476316 TI - Role of insulin and proinsulin in diabetic vascular disease. AB - Associations between loss of glucose tolerance, insulin resistance, and ischemic heart disease (IHD) are of great current concern. Considerable controversy and uncertainty relates to the mechanism(s) that underlies these associations. Whilst there is some evidence in prospective studies of an association between hyperinsulinemia and future IHD, it is by no means strong or consistent between different studies. Hypertriglyceridemia is another possible factor involved in the linkage between glucose intolerance and IHD. There is good evidence for an affect of plasma nonesterified fatty acids (NEFA) to increase hepatic output of VLDL. Insulin, contrary to some suggestions, acts to lower plasma VLDL by actions directly on hepatic output and activation of adipose tissue lipoprotein lipase, and indirectly via the hormones affect of lowering plasma NEFA. Glycosylation and oxidation of lipoproteins may enhance their atherogenic potential. It is highly probable that procoagulant changes are also important processes predisposing to IHD. Associations between plasminogen activator inhibitor-1 and insulin, intact and 32,33 split proinsulin hypertriglyceridemia, and insulin resistance have been reported, but a unifying hypothesis explaining these links remains elusive. Epidemiological studies now repeated in a number of centers have shown links between infant mortality and birth weight and risk of IHD, and between birth weight and risk of impaired glucose tolerance and non-insulin-dependent diabetes mellitus (NIDDM). It has been proposed, therefore, that impairment of fetal and infant growth may underlie the associations between loss of glucose tolerance and risk of IHD. Animal models form the basis of much current research to test this concept. PMID- 7476317 TI - Intervention in diabetic vascular disease by modulation of growth factors. AB - Several growth factors have been implicated in the derangements of cellular metabolism and proliferation that occur in diabetes, eg. kidney mesangial expansion, retinal neovascular formation, and acceleration of atherosclerosis in large vessels. These phenomena contribute to the development and progression of diabetic microvascular and macrovascular disease. Pharmacological interventions aimed at reducing growth factor alterations, among other actions in diabetic vasculopathy, include a multitude of classes of drugs, such as angiotensin converting enzyme (ACE) inhibitors, calcium antagonists, lipid-lowering drugs, and somatostatin analogs. New potential interventions, ie, antisense oligonucleotide local delivery, are being applied in growth factor research and may prove beneficial in diabetic macrovascular disease. PMID- 7476318 TI - Defining growth hormone deficiency in adults. AB - The absence of a distinct clinical syndrome calls for a strategy to reliably identify patients with hyposomatotropism. However, there is no consensus as to the most appropriate method of defining growth hormone (GH) deficiency in adults. Since GH secretion falls with senescence and is also reduced by obesity, both of these factors must be controlled for in such an evaluation. We have investigated the relative diagnostic merits of measuring (1) peak GH response to insulin induced hypoglycemia (ITT), (2) mean 24-hour GH concentration derived from 20 minute sampling (IGHC), (3) serum IGF-I levels, and (4) serum insulin-like growth factor (IGF)-binding protein-3 (IGFBP-3) levels. These tests were undertaken in 23 patients considered GH-deficient from extensive organic pituitary disease and in 35-sex-matched normal subjects of similar age and body mass index. The ITT was the only test capable of distinguishing patients with organic GH deficiency from matched normal subjects. The sensitivity of the GH radioimmunoassay (0.2 ng/mL) limited the utility of IGHC measurements, since many subjects from both groups had undetectable values. Using a GH assay with a 100-fold greater sensitivity, we found a better but still incomplete separation of values between the two groups. There was a significant overlap of IGF-I and IGFBP-3 values, with only a third of GH-deficient subjects having low IGF-I values. The limitation of IGF-I has been confirmed by others, although its sensitivity as a diagnostic test is greater in young adults. We conclude that organic GH deficiency in adults can be reliably diagnosed by the ITT.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7476319 TI - Metabolic effects of growth hormone in children. PMID- 7476320 TI - Beef tallow diet decreases norepinephrine turnover rates in rat hypothalamus and cerebral cortex. AB - The effects of dietary fats consisting of different fatty acids on norepinephrine (NE) turnover rates in various brain regions were studied in rats. Sprague-Dawley male rats were meal-fed isoenergetic diets based on safflower oil or beef tallow for 8 weeks. Body weight gain during the experimental period did not differ between the two groups of rats. The weights of brain regions were also similar in the two diet groups. Basal NE contents and turnover rates in the hypothalamus and cortex were significantly lower in rats fed the beef tallow diet versus the safflower oil diet. These results suggest that the beef tallow diet reduces noradrenergic functions in brain regions as compared with the safflower oil diet in rats. PMID- 7476321 TI - Triiodothyronine treatment increases substrate cycling between pyruvate carboxylase and malic enzyme in perfused rat liver. AB - The relative roles of pyruvate kinase and malic enzyme in substrate cycling between pyruvate and oxaloacetate were examined in perfused livers of 24-hour fasted normal and triiodothyronine (T3)-treated rats using an inhibitor of malic enzyme (hydroxymalonate). Livers were perfused for 60 minutes in a recirculating system with [3-13C]alanine (10 mmol/L, 99% 13C-enriched). The combined flux through pyruvate kinase plus malic enzyme relative to pyruvate carboxylase flux was assessed by the 13C-enrichment ratio of alanine C2 to glucose C5 in the perfusate, determined with 13C and 1H nuclear magnetic resonance (NMR) spectroscopy. In normal rat livers, the relative carbon flux through pyruvate kinase plus malic enzyme to pyruvate carboxylase was 0.18 +/- 0.04, and increased to 0.44 +/- 0.08 (P < .05) in the T3-treated group. After addition of hydroxymalonate, this relative carbon flux was unchanged in normal rat livers, but decreased to 0.15 +/- 0.04 (P < .01) in the T3-treated group, suggesting that the increased carbon flux in T3-treated livers was caused by increased flux through malic enzyme. Malic enzyme activity increased from 0.36 +/- 0.05 U/g liver in normal livers to 2.51 +/- 0.50 U/g liver (P < .05) in the T3-treated group, whereas there was no effect of T3 treatment on pyruvate kinase activity.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7476322 TI - Relationship between obesity and concentration and composition of low-density lipoprotein subfractions in normoinsulinemic men. AB - Obesity, insulin resistance (IR) with hyperinsulinemia, and a dyslipoproteinemia characterized by reduced high-density lipoprotein 2 (HDL2) cholesterol and elevated levels of small, dense low-density lipoprotein (LDL) particles are risk factors for coronary artery disease (CAD). The impact of obesity independent of hyperinsulinemia on the concentration and composition of small, dense LDL subfractions is uncertain. The aim of this study was to investigate the relationship between obesity indices, namely body mass index (BMI), skinfold measurements (SF), and waist to hip ratio (WHR), and LDL-subfraction particle concentration and composition in 200 healthy men without evidence of IR. A precise analysis of the concentration of lipids and apolipoproteins and the composition of very-low-density lipoprotein (VLDL), intermediate-density lipoprotein (IDL), and two HDL- and six LDL-subfraction particles was obtained using the technique of density-gradient ultracentrifugation. Dividing the individuals according to BMI showed that those with a BMI greater than 27 kg/m2 had significantly lower HDL2 cholesterol and apolipoprotein (apo) A-I and higher VLDL and IDL cholesterol and apo B concentrations than those with a BMI less than 25 kg/m2. Regarding LDL particles, we found that men with a BMI above 25 kg/m2 had significantly more small, dense LDL particles (d 1.044 to 1.063 g/mL) and correspondingly fewer medium, dense LDL particles (d 1.031 to 1.037 g/mL) than leaner men; those with a BMI above 27 kg/m2 had the highest concentration of circulating small, dense LDL particles. These findings were not influenced by fasting insulin concentrations, IR, or WHR.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7476323 TI - Regulation of hormone-sensitive lipase in streptozotocin-induced diabetic rats. AB - Insulin deficiency as seen in insulin-dependent diabetes mellitus causes an activation of lipolysis in adipose tissue that results in hydrolysis of stored triglycerides and release of large amounts of fatty acids into the plasma, leading to diabetic ketoacidosis (DKA). Hormone-sensitive lipase (HSL) is thought to be the rate-limiting enzyme of lipolysis in adipose tissue. This study was designed to examine the effects of insulin deficiency on the regulation of HSL in isolated adipocytes. Insulin deficiency was induced by a single dose of streptozotocin. After 8 days, some animals were treated with insulin, and all animals were killed 10 days after induction of insulin deficiency. Compared with levels in control rats, 10 days of insulin deficiency increased HSL activity twofold (P < .05), as assayed for neutral cholesterol esterase activity, and insulin treatment returned HSL activity to normal. HSL protein was increased twofold (P < .05) in streptozotocin-induced diabetic rats, as estimated by immunoblotting, but remained elevated after insulin treatment. Levels of HSL mRNA assessed by Northern blot analysis also increased twofold (P < .01) in adipose cells isolated from streptozotocin-induced diabetic rats, and remained elevated after insulin treatment. In conclusion, our studies suggest that 10 days of insulin deficiency increases HSL expression via pretranslational mechanisms and short-term insulin treatment returns HSL activity to normal via posttranslational mechanisms in adipose tissue. PMID- 7476324 TI - Insulin sensitivity, insulin secretion, and glucose effectiveness in obese subjects: a minimal model analysis. AB - The aim of the present study was to estimate insulin sensitivity (SI), insulin secretion, and glucose effectiveness in 14 obese subjects who were further divided into two groups: one with normal glucose tolerance and the other with impaired glucose tolerance (IGT). Glucose tolerance was determined by criteria of the World Health Organization. All subjects were Japanese. They underwent a modified frequently sampled intravenous glucose tolerance test: glucose (300 mg/kg body weight) was administered, and insulin (20 mU/kg body weight given over 5 minutes) was infused from 20 to 25 minutes after administration of glucose. SI and glucose effectiveness at basal insulin (SG) were estimated by Bergman's minimal model method. Body mass index (33.0 +/- 1.8 v 30.9 +/- 1.5 kg/m2, P > .05) and fasting insulin level (127.9 +/- 30.0 v 107.4 +/- 14.4 pmol/L, P > .05) were higher in obese IGT subjects than in normal obese subjects, but were not statistically significant. With regard to fasting glucose level, obese subjects with IGT (5.9 +/- 0.3 mmol/L) had significantly higher levels than those with normal glucose tolerance (5.1 +/- 0.2 mmol/L, P < .01). There was no significant difference in SI between the two groups (0.53 +/- 0.10 v 0.56 +/- 0.13 x 10( 4).min-1.pmol/L-1, P > .05). Pancreatic insulin secretion expressed as the integrated area of plasma insulin above the basal level during the first 19 minutes was significantly lower in obese subjects with IGT (3,366 +/- 1,495 pmol/L.min) than in those with normal glucose tolerance (16,400 +/- 4,509 pmol/L.min, P < .05).(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7476325 TI - Bezafibrate therapy in patients with isolated low high-density lipoprotein cholesterol levels may have a beneficial effect in prevention of atherosclerosis. AB - Although a low plasma high-density lipoprotein cholesterol (HDL-C) level is a well-accepted risk factor for coronary artery disease (CAD), it is unclear whether pharmacologic agents can effectively increase HDL-C levels and/or reduce the incidence of CAD in patients with isolated low HDL-C levels. An important determinant of HDL levels is the efficiency of postprandial lipoprotein catabolism. The purpose of the present study was to evaluate the efficacy of bezafibrate therapy in increasing HDL-C levels in these patients and to examine its effect on postprandial lipoprotein levels. Fasting and postprandial lipid and lipoprotein levels were studied in 23 patients with isolated low HDL-C levels before and during 3 and 6 months of bezafibrate treatment. Postprandial lipoprotein levels were evaluated using the vitamin A-fat loading test, in which these intestinally derived lipoproteins are specifically labeled with retinyl palmitate (RP). Patients with isolated low HDL had significantly higher levels of chylomicron RP than a control group of 19 normolipidemic subjects. The area below the chylomicron RP curve was 17,773 +/- 6,821 versus 13,936 +/- 6,217 micrograms/L.h, respectively (P < .005). No differences were found in chylomicron remnant levels between the groups. Bezafibrate therapy reduced the chylomicron RP area by 27%, from 17,773 +/- 6,821 to 12,895 +/- 2,576, and the nonchylomicron RP area by 25%, from 6,059 +/- 3,310 to 4,430 +/- 1,963 (P < .0001). It increased fasting HDL-C levels from 35 +/- 3 to 38 +/- 1.4 mg/dL after 3 months (P < .001) and to 40 +/- 2.2 mg/dL after 6 months (P < .001).(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7476326 TI - Effects of long-term treatment with low-dose pravastatin on biliary lipid and bile acid composition in patients with nonfamilial hyperlipoproteinemia. AB - We tested the possibility that pravastatin, a competitive inhibitor of hepatic hydroxymethyl glutaryl coenzyme A (HMG CoA) reductase, would alter cholesterol saturation of gallbladder bile by decreasing its cholesterol saturation index and/or degree of fatty acyl chain unsaturation in lecithin. Eighteen patients with type IIa hyperlipoproteinemia were treated with pravastatin 10 mg/d for 12 months. Gallbladder bile samples were aspirated with a duodenal tube by stimulating gallbladder contraction with intramuscular administration of cerulein before and after treatment. Serum cholesterol level was significantly reduced by 20% after 3 months, and this level was maintained after 12 months. In contrast, the cholesterol saturation index of gallbladder bile was not altered after 3 months (1.52 +/- 0.20 v 1.70 +/- 0.24), but it decreased significantly after 12 months (0.95 +/- 0.11, P < .01). The degree of fatty acyl chain unsaturation tended to decrease, although this was not statistically significant except for the decrease in molar percent of linoleate after 3 months. These findings suggest that long-term treatment with an inhibitor of HMG CoA reductase improves bile lithogenicity even at a comparatively low dose, and can decrease the incidence and complications of cholesterol gallstones. PMID- 7476327 TI - The effects of weight reduction to ideal body weight on body fat distribution. AB - Obesity is a well-known health risk factor. Several studies have demonstrated that upper-body fat distribution plays a major role in the association between increased adiposity and metabolic disorders. The present study was undertaken to evaluate changes in intraabdominal and subcutaneous fat areas in obese subjects undergoing a weight reduction to their ideal body weight (IBW), as defined by a body mass index (BMI) no greater than 21 or body fat less than 30%, and compare the fat distribution at IBW with that of never-obese control subjects. We studied 33 obese women (151% +/- 1% of IBW; BMI, 31.6 +/- 2.5 [mean +/- SE]) before and after weight loss and a control group of 16 never-obese women (101.0% +/- 1.0% of IBW; BMI, 21.2 +/- 1.1). Eighteen obese women successfully achieved and stabilized at IBW for at least 2 months. Nonsuccessful obese subjects were significantly younger than reduced-weight subjects, but other physical characteristics were similar. In obese, reduced-obese, and never-obese groups, weight was 85 +/- 2.0, 62 +/- 1, and 58 +/- 1 kg; percent body fat was 41% +/- 1%, 24% +/- 2%, and 23% +/- 1%; intraabdominal fat area was 82 +/- 5, 28 +/- 3, and 25 +/- 4 cm2; waist subcutaneous fat area was 275 +/- 15, 120 +/- 9, and 81 +/- 7 cm2; hip subcutaneous fat area was 416 +/- 17, 204 +/- 10, and 195 +/- 7 cm2; and waist to hip ratio (WHR) was 0.84 +/- 0.02, 0.77 +/- 0.01, and 0.73 +/- 0.01, respectively.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7476329 TI - Insulin resistance during euglycemic clamp studies in chronically undernourished rats with mild streptozocin diabetes. AB - Malnutrition has been shown to impair insulin sensitivity, but it is not known whether this effect has any impact on coexisting diabetes. Insulin sensitivity was therefore studied using the glucose clamp technique in rats with chronic nutritional deprivation superimposed on mild streptozocin (STZ) diabetes mellitus. In pair-feeding experiments, 4-week-old littermate rats were either allowed ad libitum access to food or restricted to 50% of ad libitum intake for 8 weeks, and were injected with STZ 40 mg/kg intraperitoneally halfway through the experiment. Fasting plasma glucose (FPG) was similar in both groups of rats, but fasting plasma insulin (FPI) was lower in the undernourished group (P = .016). Undernourished rats were significantly more insulin resistant during euglycemic hyperinsulinemia of the same degree, with glucose disposal rate being impaired by 50% as compared with that in ad libitum-fed diabetic littermates (24.4 +/- 2.8 v 51.5 +/- 4.4 mumol/kg/min, P = .0008). The insulin sensitivity index was significantly lower in the undernourished group (3.03 +/- 0.32 v 5.67 +/- 0.6, P = .0057). The results show that chronic undernutrition markedly reduces insulin sensitivity in rats with mild STZ diabetes. This is further evidence that chronic undernutrition is a deleterious modifying influence on coexisting diabetes mellitus. It suggests that the insulin resistance of malnutrition-related diabetes mellitus (MRDM) could potentially be an acquired defect mediated by the coexistent undernutrition, rather than a "distinctive" feature that is intrinsically unique to this diabetic syndrome. PMID- 7476328 TI - Hemostatic function in young subjects with central obesity: relationship with left ventricular function. AB - This study was designed to evaluate coagulation and fibrinolysis activity and their relationship with left ventricular function in young obese subjects with central fat distribution. We assessed coagulation and fibrinolysis activity by evaluation of factor VII activity, fibrinogen and plasminogen, plasminogen activator inhibitor (PAI), and tissue plasminogen activator antigen basally (tPA1) and after venous occlusion (tPA2). These measures were evaluated in young (< 40 years) obese subjects with central fat distribution (n = 19) and in comparable lean subjects (n = 20). Blood glucose, triglycerides, total and high density lipoprotein (HDL) cholesterol, apolipoprotein (apo) A1 and apo B, fasting immunoreactive insulin, and lipoprotein(a) levels were also measured by current methods. Left ventricular ejection fraction (LVEF) and peak filling rate (PFR) determined by radionuclide angiocardiography and left ventricular mass (LVM) and LVM indexed for body height (LVM/H) determined by echocardiographic study were calculated. Central obesity was evaluated by the waist to hip ratio (WHR) according to the criteria of the Italian Consensus Conference of Obesity. Factor VII (P < .001), fibrinogen (P < .001), plasminogen (P < .001), PAI activity (P < .001), tPA1 (P < .02), fasting blood glucose (P < .01), apo B (P < .02), and immunoreactive insulin (P < .01) were significantly higher in obese than in lean subjects. In contrast, HDL cholesterol (P < .01), tPA2 (P < .01), LVEF (P < .001), and PFR (P < .02) were significantly lower in obese than in lean subjects. In all subjects, WHR correlated directly with fibrinogen and inversely with tPA2; LVEF correlated inversely with tPA1, PAI, and fibrinogen; and PFR correlated inversely with factor VII activity.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7476330 TI - In vitro regulation of low-density lipoprotein receptor interaction by fatty acids. AB - Low-density lipoprotein (LDL) receptor binding is the initial step in receptor mediated clearance. Dietary fat composition is known to affect LDL clearance, but the mechanism of the effect is unknown. We have examined the effects of altered membrane fatty acid composition, as might occur when specific dietary fats are consumed, on LDL binding using a Chinese hamster ovary (CHO) line that constitutively expresses the human LDL receptor. Binding of pooled human LDL to its receptor was compared in cells enriched with various fatty acids. Binding affinity was greater (lower Kd) for cells grown in 16:0-, 18:0-, or 18:1-enriched media than for those grown in 18:2 (P < .0001). The apparent receptor number (Bmax) was lower for cells enriched in saturated fatty acids and 18:1. Fluidity was assessed by measuring diphenylhexatriene (DPH) fluorescence anisotropy (rs). Cells enriched in 18:1 or 18:2 were the most fluid (P < .003). The correlation between binding and fluidity (r = .24, P = .27) was weak and did not appear to explain the effects of fatty acid modification on LDL receptor binding. Thus, it appears that cellular enrichment in 16:0, 18:0, and 18:1 increases binding affinity by affecting properties other than membrane fluidity. Changes in Bmax may also contribute to the observed differences in LDL binding. PMID- 7476331 TI - Insulin secretion by pancreas of athymic mice injected with peripheral mononuclear cells from insulin-dependent diabetic patients. AB - We studied the effect of peripheral blood mononuclear cells (PBMNC) from insulin dependent diabetic (IDDM) children on the insulin secretion pattern of the pancreas from recipient athymic mice. PBMNC from healthy controls or IDDM patients in different stages of disease were injected into athymic mice. PBMNC from newly diagnosed IDDM children elicited basal nonfasting hyperglycemia and in vitro inhibition of the first and second phases of glucose-stimulated insulin secretion in recipient mice. Animals injected with cells from chronically IDDM children showed normoglycemia, abnormal tolerance to glucose, and inhibition of first-phase insulin secretion. Mitomycin C treatment of MNC from IDDM patients abolished insulin secretion inhibition in recipient mice. PBMNC from newly diagnosed and chronically IDDM patients showed positive anti-beta-cell cellular immune aggression. Mice injected with cells from patients during the remission period showed normoglycemia and no alteration of insulin secretion patterns. When relapsed to their former clinical stage, injection of the cells significantly inhibited first-phase glucose-induced insulin secretion in recipients. PBMNC from newly diagnosed IDDM patients were found to migrate to the pancreas of recipient mice preferably as compared with cells from controls. Cells from chronically IDDM patients cultured with concanavalin A (Con A) increased insulin secretion inhibition; despite this, cells from children during the remission period cultured with Con A failed to modify insulin secretion in recipients. These results show that injection of PBMNC from diabetic patients leads to insulin secretion impairment in recipient mice pancreas, and provide a basis for the study of mechanisms involved in the onset and modulation of anti-beta-cell cellular immune aggression induced by human PBMNC. PMID- 7476332 TI - Lean body mass in twins. AB - A study of 49 pairs of monozygous (MZ) twins and 38 pairs of same-sexed dizygous (DZ) twins showed that lean body mass (LBM), as determined by potassium 40 counting, is under genetic influence. Intrapair variances for LBM are much smaller than those for body fat, which suggests that LBM has a higher degree of heritability. There is a correlation between the magnitude of intrapair LBM differences and intrapair weight differences for both sets of twins, showing that environment is also an important influence. The effect of weight variation on LBM variation is greater for thin people than for those with appreciable burdens of body fat, an observation previously made on individuals who undergo a nutrition induced weight change. PMID- 7476334 TI - Increase of voltage-sensitive calcium channels and calcium accumulation in skeletal muscles of streptozocin-induced diabetic rats. AB - The number of voltage-sensitive calcium channels (VOCC) in triceps surae muscle membrane fractions isolated from control and streptozocin (STZ)-induced diabetic rats was determined using [3H]PN200-110, a dihydropyridine derivative, as a ligand. Furthermore, quantitative analysis of calcium in soleus muscle fibers was performed by the calcium oxalate-pyroantimonate method and x-ray microanalysis. The maximum binding (Bmax) of [3H]PN200-110 in skeletal muscle membrane isolated from 10-week diabetic rats (1,091 +/- 77 fmol/mg protein) was increased significantly by 91% as compared with the control value (572 +/- 32 fmol/mg protein), without a significant change in Kd. The increase in Bmax of [3H]PN200 110 was dependent on the duration of diabetes, and was not found until 6 weeks after STZ injection. Insulin treatment for 8 weeks after induction of diabetes normalized Bmax to the control level (583 +/- 53 fmol/mg protein). Precipitates of calcium antimonate, identified by x-ray microanalysis, were observed much more frequently in specimens from 10-week diabetic rats versus controls. The increase in the incidence of precipitates was not observed in 3-week diabetic rats and was suppressed by 8 weeks' insulin treatment. These results indicate that the number of VOCC in chronically diabetic rats was increased in the sarcolemmal membrane of skeletal muscle and that calcium was accumulated inside skeletal muscle fibers. PMID- 7476333 TI - Comparison of the effect of fluvastatin, an hydroxymethyl glutaryl coenzyme A reductase inhibitor, and cholestyramine, a bile acid sequestrant, on lipoprotein particles defined by apolipoprotein composition. AB - In a double-blind, parallel-group, randomized study, the effects of fluvastatin (FLUV) 20 and 40 mg/d on lipoprotein particle levels were compared with those of cholestyramine (CME) 16 g/d. Lipoparticles were defined by apolipoprotein composition as either those containing both apolipoprotein (apo) B and apo E or CIII (lipoprotein [Lp] E-B or Lp CIII-B) or those containing apo AI alone (Lp AI) or in association with apo AII (Lp AI-AII). After an 8-week dietary stabilization period, 100 hypercholesterolemic patients were treated with FLUV 20 mg/d for 6 weeks and 40 mg/d for an additional 6 weeks and were compared with 48 hypercholesterolemic subjects treated with CME 16 g/d. Treatment with FLUV (40 mg/d) or CME (16 g/d) for 12 weeks was associated with a significant reduction in plasma cholesterol and low-density lipoprotein (LDL) cholesterol and a significant increase in high-density lipoprotein (HDL) cholesterol. However, plasma triglyceride levels decreased following FLUV treatment, whereas they increased with CME. These changes were associated with a significant reduction in the levels of apo B (FLUV, -24%, P < .001; CME, -26%, P < .001), apo E (FLUV, 36%, P < .001; CME, -32%, P < .001), and apo CIII (FLUV, -21%, P < .001; CME, 6%, NS).(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7476335 TI - Determinants of hydroperoxide detoxification in diabetic rat intestine: effect of insulin and fasting on the glutathione redox cycle. AB - The capacity for hydroperoxide detoxification in diabetic (DM) intestine was studied in streptozocin-induced DM rats by quantification of the intestinal glutathione (GSH) redox cycle, a key cellular pathway for peroxide elimination. A role for luminal glucose in regulation of redox cycle activity was examined in insulin-treated or 24-hour-fasted DM animals. Intestinal activities of the redox enzymes, GSH peroxidase, GSSG reductase, and glucose-6-phosphate dehydrogenase (G6PD), were significantly decreased by 17 hours' insulin treatment, whereas only G6PD was decreased by fasting. Mucosal GSH levels were also markedly decreased under these conditions. These results are consistent with an overall suppression of intestinal GSH redox cycle function by short-term administration of insulin. Insulin treatment for 7 consecutive days increased hepatic G6PD activity by fourfold but was without effect on intestinal G6PD, suggesting tissue specificity in insulin regulation of G6PD. The rate of metabolism of tert-butyl hydroperoxide (tBH) in isolated enterocytes was low in the absence of substrates (0.51 +/- 0.07 nmol/10(6) cells/min) but was increased fivefold by exogenous glucose (2.70 +/- 0.11 nmol/10(6) cells/min), indicating that glucose availability is an important contributor to intestinal detoxification of toxic hydroperoxides. Collectively, the current results show that GSH redox cycle enzymes in DM intestine are under coordinate insulin control, and that this control appears to be downregulated by short-term insulin treatment. PMID- 7476336 TI - Role of cellular ribose-5-phosphate content in the regulation of 5-phosphoribosyl 1-pyrophosphate and de novo purine synthesis in a human hepatoma cell line. AB - 5-Phosphoribosyl-1-pyrophosphate (PRPP) is an important regulator of de novo purine synthesis. However, the role of ribose-5-phosphate (R5P), the precursor for PRPP, in the regulation of PRPP and de novo purine synthesis has not yet been clarified conclusively. This study was designed to clarify interrelationships between R5P content, PRPP availability, and the rate of de novo purine synthesis in the cultured human hepatoma cell line (HepG2), a plausible model for normal human hepatocytes. Increasing glucose concentration in the culture media from 0 to 10 mmol/L resulted in a 2.9-fold elevation of cellular R5P content (from 107 +/- 31 to 311 +/- 57 nmol/g protein), associated with a correlated increase of 7.14-fold in cellular PRPP availability (from 4.76 +/- 3.4 to 34 +/- 8.4 pmol/mg protein/min) and of 149-fold in the rate of de novo purine synthesis (from 55 to 8,204 dpm/mg protein/h). Plotting the rate of de novo purine synthesis versus R5P content indicates that at a wide range of R5P content, including that prevailing in hepatocytes under physiological conditions, the rate of purine synthesis depends on R5P content. A similar dependence was also demonstrated for PRPP availability. The rate of de novo purine synthesis exhibited a sigmoidal dependence on PRPP availability. The demonstration in human hepatocytes of dependence of the rate of purine synthesis on R5P content has implications concerning the pathogenesis of purine overproduction associated with several inborn and acquired conditions in man. PMID- 7476337 TI - Glycohemoglobin levels relate to the response of adipose tissue lipoprotein lipase to insulin/glucose in obese non-insulin-dependent diabetes mellitus. AB - Adipose tissue lipoprotein lipase (ATLPL) is responsible for the provision of lipoprotein-derived fatty acids to adipocytes for storage as triglycerides. Fasting ATLPL has been shown to be decreased in non-insulin-dependent diabetes mellitus (NIDDM), an insulin-resistant state. Medically uncomplicated obesity, another state of relative insulin resistance, is associated with decreased stimulation of the enzyme in response to metabolic stimuli. It was therefore hypothesized that the increased insulin resistance of NIDDM would result in an even greater defect in the response of ATLPL to insulin/glucose. Gluteal adipose tissue biopsies were performed in 13 premenopausal obese women with NIDDM, before and after 6 hours of intravenous insulin and glucose. Metabolic data from these studies were then compared with those obtained from 26 nondiabetic obese women of similar age, weight, and fasting insulin concentration (obese controls [OBC]). As expected, fasting gluteal ATLPL activity was lower in the NIDDM group than in OBC (3.7 +/- 0.9 v 11.1 +/- 1.6 nmol free fatty acids [FFA]/min/10(6) cells, P = .0003). The change in ATLPL activity (delta ATLPL) in response to a 6-hour insulin/glucose infusion was not statistically different between the two groups (2.2 +/- 1.1 v 4.7 +/- 1.2, P = .114). However, in NIDDM subjects there was a strong positive relationship between delta ATLPL and glycohemoglobin (GHb) level (r = .883, P = .0001).(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7476338 TI - Association of insulin and insulin propeptides with an atherogenic lipoprotein phenotype. AB - A characteristic lipoprotein phenotype, including hypertriglyceridemia, a low high-density lipoprotein (HDL) cholesterol concentration, and a predominance of small, dense low-density lipoprotein (LDL) particles, is linked to insulin resistance and hyperinsulinemia. Individuals with these characteristics are supposed to be at increased risk of developing coronary heart disease (CHD). To address this issue further, relations between basal and postload glucose, insulin and insulin propeptide concentrations and subfractions of apolipoprotein (apo) B containing lipoproteins were examined in 62 consecutive Swedish nondiabetic men who had experienced a first myocardial infarction before the age of 45. A total of 41 age-matched, population-based healthy men were investigated as controls. Highly specific immunoradiometric assays were used for measuring intact proinsulin and des 31,32proinsulin levels. In all, 39% of the patients were found to be glucose-intolerant, and basal and postload hyper(pro)insulinemia were characteristic features irrespective of glucose tolerance category. Hypertriglyceridemic (HTG) lipoprotein phenotypes with a low HDL cholesterol concentration dominated among the patients, and hyperinsulinemia was linked to hypertriglyceridemia and putatively atherogenic lipoprotein traits, such as increased particle numbers of small very-low-density lipoprotein (VLDL) and intermediate-density lipoprotein (IDL) and triglyceride enrichment of LDL. The corollary of these findings is that insulin resistance is a characteristic feature of young postinfarction patients and is accompanied by a complex atherogenic lipoprotein phenotype, new components of which are an abundance of small cholesteryl ester-rich VLDL and an elevated LDL triglyceride concentration. PMID- 7476339 TI - Metabolic effects of troglitazone on fat-induced insulin resistance in the rat. AB - Troglitazone is a new orally active hypoglycemic agent that has been shown to ameliorate insulin resistance and hyperinsulinemia in both diabetic animal models and non-insulin-dependent diabetes mellitus (NIDDM) subjects. To determine whether this drug could prevent the development of diet-induced insulin resistance and related abnormalities, we studied its effect on insulin resistance induced by high-fat feeding in rats. Normal male Sprague-Dawley rats were fed a high-fat diet for 3 weeks with and without troglitazone as a food mixture (0.2%) or were fed normal chow. In vivo insulin action was measured using a euglycemic hyperinsulinemic clamp at two different insulin infusion rates, 4 (submaximal stimulation) and 40 (maximal stimulation) mU/kg/min. Fat feeding markedly reduced the submaximal glucose disposal rate ([GDR], 26.4 +/- 1.3 v 37.5 +/- 1.4 mg/kg/min, P < .01) and maximal GDR (55.9 +/- 1.3 v 64.5 +/- 1.3 mg/kg/min, P < 0.5), reduced the suppressibility of submaximal hepatic glucose production ([HGP], 3.2 +/- 0.9 v 1.5 +/- 0.5 mg/kg/min, P < .05), and resulted in hyperlipidemia. Troglitazone treatment did not affect any of these parameters. Insulin resistance induced by fat feeding is the first experimental model in which troglitazone failed to correct or partially correct the insulin resistance. PMID- 7476340 TI - Zinc deficiency-induced anorexia influences the distribution of serum insulin like growth factor-binding proteins in the rat. AB - Zinc (Zn) deficiency can result in severe growth retardation in mammals, and in a number of animal model systems it leads to low circulating insulin-like growth factor-I (IGF-I) concentrations. Using a weanling male rat model and a number of feeding schemes, we show that in addition to lower circulating IGF-I concentrations, Zn deficiency leads to alterations in the distribution of serum IGF-binding proteins (IGFBPs). Serum from Zn-deficient animals labeled in vitro with [125I]IGF-I displayed three peaks of tracer activity: 150 kd (IGFBP-3), 37 kd (IGFBP-2 and -1), and 8 kd (free [125I]IGF-I). Relative to controls, Zn deficient animals demonstrated more tracer binding in the 37-kd region, whereas less was found in the 150- and 8-kd peaks. Serum from chronically calorie restricted fed animals displayed [125I]IGF-I binding profiles similar to Zn deficient serum, implicating Zn deficiency-induced anorexia as the principle factor underlying both the lower circulating IGF-I and the alterations in IGFBP profiles. Concentrations of IGFBP-4 were unaffected by diet manipulation based on sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE)/Western ligand blot (WLB) analysis. PMID- 7476341 TI - Weight loss reduces abdominal fat and improves insulin action in middle-aged and older men with impaired glucose tolerance. AB - Aging is associated with an increased accumulation of abdominal fat, glucose intolerance, and insulin resistance. We tested the hypothesis that diet-induced weight loss would reduce the abdominal distribution of fat and improve glucose tolerance and insulin action in a group of obese middle-aged and older men with normal or impaired glucose tolerance (IGT). Oral glucose tolerance tests (OGTTs) were performed at baseline and after 9 months of diet-induced weight loss in 35 men (mean age, 60 +/- 8 years). Fifteen men of comparable age and degree of obesity who did not participate in the weight loss intervention served as controls. Subjects lost 9.0 +/- 2.0 kg (mean +/- SD) body weight (P < .001), resulting in a 19% reduction in percent body fat (30.0 +/- 4.0% to 24.0% +/- 4.0%, P < .001), an 8% reduction in waist circumference (104.0 +/- 7.0 to 96.0 +/ 7.0 cm, P < .001), and a 2% reduction in waist to hip ratio [WHR] (0.97 +/- 0.06 to 0.95 +/- 0.06, P < .01). Weight loss improved glucose tolerance: nine men with IGT at baseline reverted to normal glucose tolerance following the intervention. Glucose area during the OGTT was significantly reduced after weight loss (-22.0%, P < .001), while it increased in control subjects (+32%, P < .004). In multiple regression analysis, the improvement in glucose area following weight loss in these 35 men was attributed to the reduction in waist circumference (P < .01) and baseline glucose area (P < .05).(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7476342 TI - Acipimox potentiates growth hormone response to growth hormone-releasing hormone by decreasing serum free fatty acid levels in hyperthyroidism. AB - Hyperthyroidism is associated with an impairment of growth hormone (GH) responses to secretagogues. The aim of this study was to evaluate the effect of acipimox, an antilipolytic agent able to decrease free fatty acids (FFA), on GH response to GH-releasing hormone (GHRH) in hyperthyroid and normal control subjects. We studied six men with hyperthyroidism; seven normal men served as control subjects. Each subject underwent treatment with (1) 2 tablets of placebo orally or (2) 500 mg acipimox orally, 120 minutes before intravenous (IV) injection of 1 microgram/kg GHRH-(1-29)NH2. GH response to GHRH in hyperthyroid patients was markedly reduced; the mean peak GH response (9.6 +/- 1.0 microgram/L) and the area under the GH response curve (12.9 +/- 1.3 micrograms/L x 2 h) were lower than those of control subjects (25.7 +/- 1.8 micrograms/L, P < .05; 28.7 +/- 2.1 micrograms/L x 2 h, P < .05). Hyperthyroid patients had higher baseline levels of plasma FFA than control subjects (998.0 +/- 38.9 v 498.0 +/- 36.0 muEq/L, P < .01). Acipimox decreased FFA levels in both hyperthyroid and control subjects; the lowest FFA levels of hyperthyroid subjects induced by acipimox were similar to those of control subjects. After acipimox pretreatment, GH responses to GHRH increased significantly (P < .05); the mean peak plasma GH level (25.9 +/- 4.6 micrograms/L) was similar to the peak GH levels of control subjects during the GHRH test, and the area under the GH response curve (41.1 +/- 6.7 micrograms/L x 2 h) was even higher than that of control subjects with the GHRH test.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7476343 TI - Promotion of cyst(e)ine uptake. PMID- 7476344 TI - Ovothiols. PMID- 7476345 TI - Characterization of the cysteine-rich zinc-binding domains of protein kinase C by X-ray absorption spectroscopy. PMID- 7476346 TI - Modifications of cysteine-rich regions in protein kinase C induced by oxidant tumor promoters and enzyme-specific inhibitors. PMID- 7476347 TI - Intracellular distribution of thiols and disulfides: assay of mitochondrial glutathione transport. PMID- 7476348 TI - Glutathione and protein kinase C in peripheral nervous tissue. PMID- 7476349 TI - Expression of protein kinase C gamma regulatory domain elements containing cysteine-rich zinc-coordinating regions as glutathione S-transferase fusion proteins. PMID- 7476350 TI - Redox regulation of activation of NF-kappa B transcription factor complex: effects of N-acetylcysteine. PMID- 7476351 TI - Redox regulation of DNA-protein interactions by biothiols. PMID- 7476352 TI - Suppression of protooncogene c-fos expression by antioxidant dihydrolipoic acid. PMID- 7476354 TI - Thioredoxin and thioredoxin reductase. PMID- 7476353 TI - Dihydrolipoamide dehydrogenase: activity assays. AB - We have described the most commonly used assay procedures determination of the DHLipDH activities from prokaryotic and eukaryotic cells. We have also described the procedures for the preparation of tissue extracts to determine the enzymatic activity. We have briefly alluded to the problems inherent in these assay procedures and suggested some precautions for using the assays to determine DHLipDH activity. We have also included a statement about the purification procedures and described some of the properties of this enzyme, which might help the reader in planning studies. We have, by no means, attempted to be comprehensive in this regard; therefore, the reader should consult the appropriate references (and references therein). PMID- 7476355 TI - Selenite and selenodiglutathione: reactions with thioredoxin systems. PMID- 7476357 TI - Thioredoxin and seed proteins. PMID- 7476356 TI - Measurement of equilibrium midpoint potentials of thiol/disulfide regulatory groups on thioredoxin-activated chloroplast enzymes. PMID- 7476358 TI - Analysis and manipulation of target enzymes for thioredoxin control. PMID- 7476359 TI - Effects of thioredoxin on activation of transcription factor NF-kappa B. PMID- 7476360 TI - Glutathione biosynthesis and its inhibition. PMID- 7476361 TI - Thioredoxin genetics. PMID- 7476362 TI - Ferredoxin: thioredoxin system. PMID- 7476363 TI - Glutaredoxin. PMID- 7476364 TI - Nuclear magnetic resonance of thioredoxin and glutaredoxin. PMID- 7476365 TI - Glutathione distribution in natural products: absorption and tissue distribution. PMID- 7476366 TI - Glutathione: dehydroascorbate oxidoreductases. PMID- 7476367 TI - Selenophosphate. PMID- 7476368 TI - Anethole dithiolethione: biochemical considerations. PMID- 7476369 TI - Use of linoleic acid radiolysis for assay of antioxidant action of anethole dithiolethione. PMID- 7476371 TI - Ebselen. PMID- 7476370 TI - Cold light fluorimetry: a microtitration technology for cell culture to evaluate anethole dithiolethione and other biothiols. PMID- 7476372 TI - Thiol compounds and adult T-cell leukemia virus infection: a potential therapeutic approach. PMID- 7476373 TI - Diversity of glutathione peroxidases. PMID- 7476374 TI - Purification of soluble human glutathione S-transferases. PMID- 7476375 TI - Measurement of gamma-glutamyl transpeptidase and gamma-glutamylcysteine synthetase activities in cells. PMID- 7476376 TI - Thiol transport from human red blood cells. PMID- 7476377 TI - Glutathione mutants. PMID- 7476378 TI - Cystathionine. PMID- 7476379 TI - Specificity mapping of bacterial lectins by inhibition of hemagglutination using deoxy and deoxyfluoro analogs of receptor-active saccharides. PMID- 7476381 TI - Association of bacteria with human phagocytes. PMID- 7476380 TI - Use of synthetic peptides in characterization of microbial adhesins. AB - The characterization of microbial adhesins is greatly facilitated by the use of synthetic peptides. Synthetic peptides can be used to identify specific antigenic epitopes, to delineate receptor-binding domain of adhesins, and to facilitate the characterization of the adhesin, and they allow for a direct examination of structure-binding relationships. PMID- 7476382 TI - Identifying bacterial receptor proteins and quantifying strength of interactions they mediate. PMID- 7476384 TI - Use of confocal microscopy in studying bacterial adhesion and invasion. PMID- 7476386 TI - Kinetic analysis of microbial adhesion. PMID- 7476383 TI - Electron microscopical methods in adhesion. PMID- 7476385 TI - Epidemiological considerations in studies of microbial adhesion. PMID- 7476387 TI - Strategies for studying bacterial adhesion in vivo. PMID- 7476388 TI - Assay for adhesion of host cells to immobilized bacteria. PMID- 7476389 TI - Strategies for employing molecular genetics to study tip adhesions. PMID- 7476390 TI - Use of TnphoA and T7 RNA polymerase to study fimbrial proteins. PMID- 7476391 TI - Molecular cloning of adhesion genes. PMID- 7476393 TI - Interaction of bacteria with mast cells. PMID- 7476392 TI - Molecular analysis of Streptococcus pyogenes adhesion. PMID- 7476395 TI - Bacterial adhesion to and penetration of intestinal mucus in vitro. PMID- 7476394 TI - Adhesion of microbial pathogens to leukocyte integrins: methods to study ligand mimicry. PMID- 7476397 TI - Methods for studying adhesion of diarrheagenic Escherichia coli. PMID- 7476396 TI - Identification of intestinal receptors for enterotoxigenic Escherichia coli. PMID- 7476399 TI - In vitro adhesion of bacteria to exfoliated uroepithelial cells: criteria for quantitative analysis. PMID- 7476398 TI - Adhesion properties of Helicobacter pylori. PMID- 7476400 TI - Adhesion of mycoplasmas. PMID- 7476401 TI - Adhesion of oral bacteria to soft tissue. PMID- 7476402 TI - Coaggregations among oral bacteria. AB - The oral bacterial community appears to use coaggregation as a major mechanism for interbacterial adhesion and colonization of the host. Methods for measuring and evaluating the specificity of adhesion vary from the visual observation of the phenomenon to quantitative analyses. Not only is aggregation specificity reflected in the choice of partners but also in the fact that many are inhibited by galactosides and sialic acid. Each coaggregation between any two partners within a multigeneric coaggregate is independent of the others and can be shown to be distinct by using the radioactivity-based assay. By using the visual assay, it has been shown that members of the 17 genera of most frequently isolated oral bacteria exhibit coaggregation. With the exception of oral streptococci and a few oral actinomyces, the 17 genera do not exhibit intrageneric coaggregation. As a dynamic population, oral bacteria are in a constant flux of accretion and detachment, which are coupled to growth and death. This ecological community is amenable to intensive study, and the coaggregation assays described here are particularly suited to enhance progress in this study. PMID- 7476403 TI - Identifying and isolating fimbrial-associated adhesins of oral gram-negative bacteria. PMID- 7476404 TI - Analysis for adhesins and specific cytoadhesion of Trichomonas vaginalis. PMID- 7476405 TI - Adhesion of fungi. PMID- 7476406 TI - Galactose-specific adhesion mechanisms of Entamoeba histolytica: model for study of enteric pathogens. PMID- 7476407 TI - Erythrocytes as target cells for testing bacterial adhesins. PMID- 7476408 TI - Adhesion and interaction of Candida albicans with mammalian tissues in vitro and in vivo. PMID- 7476409 TI - Use of flow chamber devices and image analysis methods to study microbial adhesion. PMID- 7476411 TI - Adhesin-dependent isolation and characterization of bacteria from their natural environment. PMID- 7476410 TI - Methods for studying microbial colonization of plastics. PMID- 7476413 TI - Adhesion of urogenital organisms to polymers and prosthetic devices. PMID- 7476412 TI - Binding of extracellular matrix proteins by microbes. PMID- 7476414 TI - Bacterial adhesion measured by growth of adherent organisms. PMID- 7476415 TI - Enzyme-linked immunosorbent-based adhesion assays. PMID- 7476416 TI - Analysis of ocular microbial adhesion. PMID- 7476417 TI - Bacterial adhesion to hydroxylapatite. PMID- 7476418 TI - Measurement of microbial adhesion to hydrophobic substrata. PMID- 7476419 TI - Isolation and structural characterization of adhesin polysaccharide receptors. AB - The procedure for the purification of the adhesin polysaccharide receptor and its hexasaccharide repeating unit from whole S. oralis ATCC 55229 by chemical, enzymatic, and chromatographic techniques has been described. Chemical, chromatographic, and mass spectrometric procedures allow preliminary structural characterization of the hexasaccharide repeating unit and polysaccharide. The structural characterizations of the hexasaccharide and polysaccharide are completed using several 1D and 2D NMR techniques. Identification of the anomeric 1H and 13C signals of the glycosyl residues permits, by virtue of their chemical shifts and coupling constants (3JHH and 1JCH), the determination of the configurations of the glycosidic linkages. The HMBC connectivities permit the establishment of the hexasaccharide sequence as Rhap alpha(1-->2)Rhap alpha(1- >3)Galp alpha(1-->3)Galp beta(1-->4)Glcp beta(1-->3)Gal. The 1H NMR chemical shifts of the polysaccharide, as determined by the combination of COSY and TOCSY experiments, and the observed interglycosidic NOESY cross-peaks reveal the structure of the polysaccharide to be [formula: see text] where the position of the glycerol (Gro) phosphate moiety has been determined by [1H, 31P] NMR spectroscopy. PMID- 7476420 TI - Identification of receptors for bacterial lectins by blotting techniques. PMID- 7476421 TI - Purification and characterization of galactose- and N-acetylgalactosamine specific adhesin lectin of Entamoeba histolytica. PMID- 7476422 TI - Purification and GTPase-activating protein activity of baculovirus expressed p190. PMID- 7476423 TI - Purification of baculovirus-expressed Cdc42Hs. PMID- 7476424 TI - GTPase-activating protein activity of n(alpha 1)-Chimaerin and effect of lipids. PMID- 7476425 TI - Characterization of breakpoint cluster region kinase and SH2-binding activities. AB - BCR is an interesting signaling protein, whose cellular function is currently unknown. Its biochemical properties include serine kinase activity, SH2-binding activity, and a GTPase-activating activity. The SH2-binding activity is particularly interesting because it may link BCR to signaling pathways involving SH2-containing molecules. Since tyrosine phosphorylation of BCR has been detected in CML-derived cell lines and since tyrosine-phosphorylated BCR shows increased affinity toward certain SH2 domains, it seems particularly important to further characterize this activity. This chapter described a simple purification scheme for partial purification of BCR, which can be used to assess in vitro kinase and SH2-binding activities. PMID- 7476426 TI - Identification of GTPase-activating proteins by nitrocellulose overlay assay. PMID- 7476427 TI - Identification of 3BP-1 in cDNA expression library by SH3 domain screening. PMID- 7476428 TI - Purification and properties of Rac2 from human leukemia cells. PMID- 7476429 TI - Serum induction of RhoG expression. PMID- 7476430 TI - Microinjection of epitope-tagged Rho family cDNAs and analysis by immunolabeling. PMID- 7476431 TI - Purification and assay of recombinant C3 transferase. PMID- 7476432 TI - In vitro ADP-ribosylation of Rho by bacterial ADP-ribosyltransferases. PMID- 7476433 TI - Preparation of native and recombinant Clostridium botulinum C3 ADP ribosyltransferase and identification of Rho proteins by ADP-ribosylation. PMID- 7476434 TI - In vitro binding assay for interactions of Rho and Rac with GTPase-activating proteins and effectors. PMID- 7476435 TI - Purification and assay of kinases that interact with Rac/Cdc42. PMID- 7476436 TI - Yeast two-hybrid system to detect protein-protein interactions with Rho GTPases. PMID- 7476437 TI - Assay for Rho-dependent phosphoinositide 3-kinase activity in platelet cytosol. PMID- 7476438 TI - Neutrophil phospholipase D: inhibition by Rho-GDP dissociation inhibitor and stimulation by small GTPase GDP dissociation stimulator. PMID- 7476439 TI - Purification of Rac2 from human neutrophils. PMID- 7476440 TI - Measurement of Rac translocation from cytosol to membranes in activated neutrophils. PMID- 7476441 TI - Reconstitution of cell-free NADPH oxidase activity by purified components. PMID- 7476442 TI - Genetic and biochemical analysis of Cdc42p function in Saccharomyces cerevisiae and Schizosaccharomyces pombe. PMID- 7476443 TI - Lymphocyte aggregation assay and inhibition by Clostridium botulinum C3 ADP ribosyltransferase. PMID- 7476444 TI - Inhibition of p21 Rho in intact cells by C3 diphtheria toxin chimera proteins. PMID- 7476445 TI - Purification of recombinant Rho/Rac/G25K from Escherichia coli. PMID- 7476446 TI - Growth factor-induced actin reorganization in Swiss 3T3 cells. PMID- 7476447 TI - Microinjection of Rho and Rac into quiescent Swiss 3T3 cells. PMID- 7476448 TI - Inhibition of lymphocyte-mediated cytotoxicity by Clostridium botulinum C3 transferase. PMID- 7476449 TI - Neutrophil chemotaxis assay and inhibition by C3 ADP-ribosyltransferase. PMID- 7476450 TI - Purification of Rac-GDP dissociation inhibitor complex from phagocyte cytosol. PMID- 7476451 TI - Cell motility assay and inhibition by Rho-GDP dissociation inhibitor. PMID- 7476452 TI - Cell transformation by dbl oncogene. PMID- 7476453 TI - Inhibition of Rac function using antisense oligonucleotides. PMID- 7476455 TI - Prenylcysteine-directed carboxyl methyltransferase activity in human neutrophil membranes. PMID- 7476454 TI - Purification and properties of recombinant Rho-GDP dissociation inhibitor. PMID- 7476456 TI - Measurement of intrinsic nucleotide exchange and GTP hydrolysis rates. PMID- 7476457 TI - Guanine nucleotide exchange catalyzed by dbl oncogene product. PMID- 7476458 TI - Stimulation of nucleotide exchange on Ras- and Rho-related proteins by small GTP binding protein GDP dissociation stimulator. PMID- 7476459 TI - Interaction of ect2 and Dbl with Rho-related GTPases. PMID- 7476460 TI - Solubilization of Cdc42Hs from membranes by Rho-GDP dissociation inhibitor. PMID- 7476461 TI - A general model of drug prescription. AB - Currently, there is no widely accepted structured representation of drug prescription. Nevertheless, a structured representation is required for entering and storing drug prescriptions avoiding free text in computerized systems, and for drug prescription reviews. Derived from part of the work of the European OPADE project, we describe an object-oriented model of drug prescription which incorporates important concepts such as the phase and triggering event concepts. This model can be used to record all drug prescriptions, including infusions, in a structured way avoiding free text. The phase concept allows the storage of sequentially ordered dosage regimens for a drug within the same prescription. The prescription triggering event concept allows recording of the administration of a drug conditional to dates, symptoms and clinical signs, medical procedures, and everyday life events. This model has been implemented within the OPADE project; the corresponding aspects of the user interface are presented to show how this model can be used in practice. Even if other new attributes may be added to the described objects, the structure of this model is suitable for general use in software which requires the entry, storage and processing of drug prescriptions. PMID- 7476462 TI - Modeling drug information for a prescription-oriented knowledge base on drugs. AB - There exists little theoretical analysis how to represent knowledge on drugs required for computerized drug-prescription applications. A work package drug information modeling is described which was part of the European OPADE project. We describe the content and structure of a Drug Knowledge Base (DKB) designed to meet the requirements of decision-support systems in the domain of drug therapy, and to facilitate data transfer from various information sources. The definition of the DKB content is derived from the analysis of information requirements at the various stages of the process of the clinical usage of drugs (prescribing, administration, and follow-up). The DKB structure results from the classification of the various data items along two dimensions: (1) entities in the pharmaco therapeutic domain for which information must be defined (the PharmacoTherapeutic Group, the Component, the ManufacturedPreparation, and the Presentation), and (2) the validity score of the pharmaco-therapeutic information (international, national, or local). PMID- 7476463 TI - A graphical query generator for clinical research databases. AB - Clinical research involves recording, storage and retrieval of disease-related patient data, typically using a database system. In order to facilitate ad hoc queries to clinical databases we have developed a query generator with a graphical interface. The query generator uses an object-oriented data model which is visualized by directed graphs. The main focus of our work was the definition of object-oriented user views to the partly complex data structures of a relational database. Furthermore, we tried to define graphical abstractions for all common types of queries. Thus, even for non-expert database users such as clinicians, it is easy to assemble highly complex queries for a thorough examination of the content of large research databases. PMID- 7476464 TI - Experiences using New Zealand's hospital based surveillance system for injury prevention research. AB - The focus of this paper is the Injury Prevention Research Unit's (IPRU's) experience in analysing New Zealand's national impatient public hospital injury data set. The existence of the national inpatient data management system has enabled the IPRU to develop an injury morbidity data set for the period of 1979 1992. The IPRU, thus, has data on ove 250,000 injury events that were serious enough to warrant admission to a hospital. This data set has been used extensively by the IPRU to address a wide range of injury issues from demographic, environment, activity, product, and injury perspectives. Apart from the demographic variables, those variables that have proved most useful in our work have been: length of stay, readmission indicator, a personal identifier code number, WHO International Classification of Disease coding for diagnoses and external cause of injury (E-code), and written descriptions of external cause of injuries and location of injury event. Practical examples of IPRU's use of each of these variables are given. These examples demonstrate how invaluable New Zealand's inpatient injury data set is for documenting resource utilisation, accurately determining the incidence of events, undertaking analytical epidemiological studies including evaluations, and addressing shortcomings in E codes. A key aspect of the system is the narrative information. Evidence is produced demonstrates that the electronic recording of narratives of the circumstances of injury is an invaluable tool for conducting epidemiological research which has direct implications for injury prevention policy and practice.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7476465 TI - Automated coding of patient discharge summaries using conceptual graphs. AB - In medicine, as in other domains, indexing and classification is a natural human task which is used for information retrieval and representation. In the medical field, encoding of patient discharge summaries is still a manual time-consuming task. This paper describes an automated coding system of patient discharge summaries from the field of coronary diseases into the ICD-9-CM classification. The system is developed in the context of the European AIM MENELAS project, a natural-language understanding system which uses the conceptual-graph formalism. Indexing is performed by using a two-step processing scheme; a first recognition stage is implemented by a matching procedure and a secondary selection stage is made according to the coding priorities. We show the general features of the necessary translation of the classification terms in the conceptual-graph model, and for the coding rules compliance. An advantage of the system is to provide an objective evaluation and assessment procedure for natural-language understanding. PMID- 7476466 TI - A model for medical knowledge representation application to the analysis of descriptive pathology reports. AB - A new knowledge-representation system is presented, designed for medical knowledge-based applications and in particular for the analysis of descriptive medical reports. Knowledge is represented at two levels. A definitional level uses a concept-type hierarchy, a relation-type hierarchy, and a set of schematic graphs to define the concepts used and the relations between them, as well as different types of cardinality restrictions on these relations. A set of compositional hierarchies using the classic "has-part" relation as well as a new set-inclusion relation allows concept composition to be precisely defined. An assertional level allows the creation and manipulation of empirical data, in the form of graphs using the concepts, relations, and constraints defined at the definition level. The use of cardinality constraints in graph unification is considered in the context of descriptive medical discourse analysis. PMID- 7476468 TI - Computer-aided diagnosis and medical decision support are not synonymous. PMID- 7476467 TI - Evaluating four diagnostic methods with acute abdominal pain cases. AB - Contemporary work in medical decision support is characterized by a multitude of methods. To investigate their relative strengths and weaknesses, we built four diagnostic expert systems based on different methods (Bayes, case-based classification, heuristic classification) for analysis of the same set of 1254 cases of acute abdominal pain previously documented in a prospective multicenter study. The results of the comparative evaluation indicate that differences in overall performance are relatively small (statistically not significant). The performance depends more on the quality of the knowledge base and the case data than on the inference methods of the expert systems. Methods relying exclusively on empirical knowledge (Bayes, case-based classification) tend to have slightly higher overall performance scores due to a diagnostic bias toward ordinary and common diseases. By contrast, methods operating with expert knowledge (e.g., heuristic classification) perform slightly worse overall, but are more sensitive toward uncommon (serious) diseases. PMID- 7476469 TI - The feasibility and accuracy of anonymized record linkage to estimate shared clientele among three health and social service agencies. AB - This study was designed to test the effectiveness of three computerized methods of record linkage, using minimal identifiers. The goal of the linkage was to determine the proportion of persons receiving Public Health Nursing Services (n = 5,749) who also receive income assistance from the region (n = 38,800) or the province (n = 16,741). The first linkage method was based on full agreement of a number of variables (deterministic matching) and the second was based on the likelihood of the match (probabilistic matching). The third method combined information from methods 1 and 2. Method 3 proved to be the most effective linkage technique, with a specificity of 0.98 and sensitivity of 0.94. The linkage showed that only 17% of the Public Health clientele received income assistance from the region or province, much lower than expected. Factors which have probably led to an underestimate are discussed. PMID- 7476470 TI - A three-level graph-based model for the management of hospital information systems. AB - Information processing in hospitals, especially in university hospitals, is currently faced with two major issues: low-cost hardware and progress in networking technology leads to a further decentralization of computing capacity, due to the increasing need for information processing in hospitals and due to economic restrictions, it is necessary to use commercial software products. This leads to heterogeneous hospital information systems using a variety of software and hardware products, and to a stronger demand for integrating these products and, in general, for a dedicated methodology for the management of hospital information systems to support patient care and medical research. We present a three-level graph-based model (3LGM) to support the systematic management of hospital information systems. 3LGM can serve as a basis for assessing the quality of information processing in hospitals. 3LGM distinguishes between a procedural level for describing the information procedures (and their information interchange) of a hospital information system and thus its functionality, a logical too level, focusing on application systems and communication links, and a physical tool level with physical subsystems (e.g., computer systems) and data transmission. The examples that are presented have been taken from the Heidelberg University Hospital Information System. PMID- 7476471 TI - A quantitative ranking of the Biomedical Informatics serials. AB - We have developed a quantitative serial ranking system based on multiple citation analysis techniques, library use statistics, expert opinion, and selected distinguishing publication characteristics. Evaluation criteria categories include: average Science Citation Index (Impact Factor, Immediacy Index, Total citations) rankings from 1987 to 1992; citation source counts of multiple "core" biomedical informatics publications; a questionnaire sent to American College of Medical Informatics Fellows; publication delay; distinguishing characteristics (e.g., subscription cost, total circulation, year established, places indexed, affiliation with a professional society, major biomedical resource library holdings); and the total number of interlibrary loan requests to the U. S. National Library of Medicine. The top serials were Computers and Biomedical Research, MD Computing, Methods of Information in Medicine, Medical Decision Making and Computers in Biology and Medicine. PMID- 7476472 TI - Electrophoretic separation of dynein heavy chains. PMID- 7476473 TI - Two-dimensional separation of axonemal proteins. PMID- 7476474 TI - Analysis of flagellar calcium-binding proteins. PMID- 7476475 TI - Phosphorylation of Chlamydomonas flagellar proteins. PMID- 7476476 TI - In situ glycosylation of flagellar lipids. PMID- 7476477 TI - Isolation of cilia from Tetrahymena thermophila. PMID- 7476478 TI - Detection of flagellar protein kinases on polyvinylidene difluoride membranes following sodium dodecyl sulfate-polyacrylamide gel electrophoresis. PMID- 7476479 TI - Measurement of ATPase activity using [gamma-32P]ATP. PMID- 7476480 TI - Nonradioactive method for ATPase assays. PMID- 7476481 TI - Immunoelectron microscopy. PMID- 7476482 TI - Immunofluorescence microscopy of cilia and flagella. PMID- 7476484 TI - Scanning transmission electron microscopic analysis of the isolated dynein ATPase. PMID- 7476483 TI - Preparation of cilia and subciliary fractions from Paramecium. PMID- 7476485 TI - High-resolution negative staining of the isolated dynein ATPase. PMID- 7476486 TI - Computer-assisted analysis of flagellar structure. PMID- 7476487 TI - Preparation of cilia and flagella for thin-section transmission electron microscope analysis. PMID- 7476488 TI - Fixation of mammalian spermatozoa for electron microscopy. PMID- 7476489 TI - Reactivation of Chlamydomonas cell models. PMID- 7476490 TI - Reactivation of extracted Paramecium models. PMID- 7476491 TI - Isolation and reactivation of newt lung respiratory cilia. PMID- 7476493 TI - High-speed video microscopy of flagella and cilia. PMID- 7476492 TI - Reactivation of motility of demembranated sea urchin sperm flagella. PMID- 7476494 TI - Micromanipulation of sperm and other ciliated or flagellated single cells. PMID- 7476495 TI - Isolation of Euglena flagella. PMID- 7476496 TI - Preparation and reactivation of demembranated, cytosol-free ram spermatozoa. PMID- 7476498 TI - High-resolution imaging of flagella. PMID- 7476497 TI - Microscopic assays of flagellar dynein activity. PMID- 7476499 TI - Flagellar surface motility: gliding and microsphere movements. PMID- 7476500 TI - Assay of Chlamydomonas phototaxis. PMID- 7476501 TI - Quantification of ciliary beat frequency and metachrony by high-speed digital video. PMID- 7476502 TI - Flagellar amputation and regeneration in Chlamydomonas. PMID- 7476503 TI - Isolation and fractionation of the Tetrahymena cytoskeleton and oral apparatus. PMID- 7476504 TI - Release of the cytoskeleton and flagellar apparatus from Chlamydomonas. PMID- 7476505 TI - Isolation of ram sperm flagella. PMID- 7476506 TI - Purification of SF-assemblin. PMID- 7476507 TI - Purification of basal bodies and basal body complexes from Chlamydomonas reinhardtii. PMID- 7476508 TI - Preparation and reactivation of Spermatozopsis cytoskeletons. PMID- 7476509 TI - Centrin-based contractile fibers: chromatographic purification of centrin. PMID- 7476510 TI - Isolation of flagellar paraxonemal rod proteins. PMID- 7476511 TI - Preparation of ciliary and flagellar remnants. PMID- 7476512 TI - In vitro polymerization of tubulin from echinoderm sperm flagellar microtubules. PMID- 7476513 TI - Isolation of molluscan gill cilia, sperm flagella, and axonemes. PMID- 7476514 TI - Methods for the isolation of tektins and Sarkosyl-insoluble protofilament ribbons. PMID- 7476515 TI - Isolation of radial spoke heads from Chlamydomonas axonemes. PMID- 7476516 TI - Isolation of the dense fibers of mammalian sperm flagella. PMID- 7476517 TI - Isolation of the fibrous sheath of mammalian sperm flagella. PMID- 7476518 TI - Isolation and fractionation of ciliary membranes from Tetrahymena. PMID- 7476519 TI - Measurement of membrane potential and Na+ and H+ transport in isolated sea urchin sperm flagella and their membrane vesicles. PMID- 7476520 TI - Nonradioactive methods for labeling and identifying membrane surface proteins. PMID- 7476521 TI - Fluorescence labeling of flagellar membranes. PMID- 7476522 TI - Electrophysiology of ciliates. PMID- 7476523 TI - Isolation of Chlamydomonas mastigonemes. PMID- 7476524 TI - Isolation of flagella and their membranes from sea urchin spermatozoa. PMID- 7476525 TI - Ciliary membrane tubulin: isolation and fractionation. PMID- 7476526 TI - Peptide receptors in sea urchin sperm flagella. PMID- 7476527 TI - Assaying protein phosphatases in sperm and flagella. PMID- 7476528 TI - Flagellar adenylyl cyclases in Chlamydomonas. PMID- 7476529 TI - Isolation and characterization of sea urchin flagellar creatine kinase. AB - Phosphagen kinases play a role in metabolic function in diverse species (Bessman and Carpenter, 1985). Enzyme assays and inhibitors have been used successfully to demonstrate the role of one of these enzyme families in sea urchins. Although arginine kinase was not purified, its enzymatic activity has been detected in sea urchin eggs and embryos and many other species by simply substituting phosphoarginine (PArg) for PCr (Fujimaki and Yanagisawa, 1978; Tombes and Shapiro, 1989). ArgK does not show sensitivity to FDNB, but CrK proteins of the appropriate molecular weight were detected with this reagent in cells from many species (Tombes and Shapiro, 1989). The use of these techniques to identify and study function of phosphagen kinases in diverse species has much potential. PMID- 7476530 TI - Isolation of fish sperm flagella. PMID- 7476532 TI - Isolation of inner- and outer-arm dyneins. PMID- 7476531 TI - Isolation and characterization of protein kinases from Paramecium cilia. PMID- 7476533 TI - Separation of dynein species by high-pressure liquid chromatography. PMID- 7476534 TI - Reconstitution of dynein arms in vitro. PMID- 7476535 TI - Inhibitors of the dynein ATPase and ciliary or flagellar motility. PMID- 7476536 TI - Vanadate-mediated photolysis of dynein heavy chains. PMID- 7476537 TI - Modified nucleotides as probes for dynein. PMID- 7476538 TI - Transformation of Chlamydomonas reinhardtii. PMID- 7476539 TI - Insertional mutagenesis and isolation of tagged genes in Chlamydomonas. PMID- 7476540 TI - Molecular mapping of genes for flagellar proteins in Chlamydomonas. PMID- 7476541 TI - Mating and tetrad analysis in Chlamydomonas reinhardtii. PMID- 7476542 TI - Strategies for isolation of flagellar motility and assembly mutants in Chlamydomonas. PMID- 7476543 TI - Epitope tagging of flagellar proteins. PMID- 7476544 TI - Enrichment of mRNA encoding flagellar proteins. PMID- 7476545 TI - DNA-mediated transformation in Tetrahymena. PMID- 7476546 TI - Ciliogenesis of rat tracheal epithelial cells in vitro. PMID- 7476547 TI - Strategies for the isolation of ciliary motility and assembly mutants in Tetrahymena. PMID- 7476548 TI - Identification of new dynein heavy-chain genes by RNA-directed polymerase chain reaction. PMID- 7476550 TI - Induction of ciliogenesis in oviduct epithelium. PMID- 7476549 TI - Preparation of explant and organ cultures and single cells from airway epithelium. PMID- 7476551 TI - Preparation of teleost rod inner and outer segments. PMID- 7476552 TI - Large-scale isolation of Chlamydomonas flagella. PMID- 7476553 TI - Isolation of respiratory cilia. PMID- 7476554 TI - Serratia marcescens adherence: the effect of O-antigen presence. AB - The presence of O-antigen affects the adhesion of Serratia marcescens cells to inert and biological surfaces. Mutants defective in the O-side chain showed significant decreases in their ability to adhere to plastic and glass surfaces and to the surfaces of Foley urinary catheters, compared with their respective parental strains. Similar results were obtained when the ability to adhere human uroepithelial cells was tested. PMID- 7476555 TI - Microflora of murcha: an amylolytic fermentation starter. AB - Murcha is a traditional starter, used commonly in Darjeeling hills and Sikkim in India to ferment a variety of starchy substrates in order to produce sweet-sour alcoholic beverages, called jnards. Murcha cakes are mildly acidic (pH 5.2) and contain 13% w/w moisture and 0.7% w/w ash (dry weight basis). A total of 194 bacterial, 190 yeast and 80 mould strains were isolated from 30 samples of murcha. The counts (cfu/g fresh weight) of micro-organisms in the samples were 2.0 x 10(7) to 4.2 x 10(8) for Pediococcus pentosaceus, 4.0 x 10(7) to 6.8 x 10(8) for Saccharomycopsis fibuligera, 2.0 x 10(6) to 7.2 x 10(7) for Pichia anomala, 1.0 x 10(6) to 4.1 x 10(7) for Mucor circinelloides and < 10 to 1.0 x 10(6) for Rhizopus chinensis. While all the species were prevalent in 100% of the samples, R. chinensis was detected in only 50% of them. Both the moulds but one yeast species, S. fibuligera had amylolytic activity. PMID- 7476556 TI - Lethal effect of carbonyl cyanide m-chlorophenylhydrazone on Escherichia coli and a halotolerant Brevibacterium species. AB - An attempt was made to examine quantitatively the survival of Escherichia coli and the halotolerant Brevibacterium species, as a function of the exposure time to carbonyl cyanide m-chlorophenylhydrazone (CCCP), a proton conductor. Growth rates, viability, and protein concentrations of E. coli grown in the absence of glucose were unaffected by the addition of 100 microM CCCP. In the presence of glucose the viability was reduced after 24 h incubation with CCCP. Such a high efficiency of CCCP lethality for E. coli cells in the presence of glucose was attributed to not only the growth phase but also the acidic pH of the culture due to metabolites from glucose, mainly lactic acid. The culture of E. coli appeared to be in a syncopic state hovering between life and death when it was exposed to CCCP in the presence of more than 30 mM glucose. In contrast, growth rates of Brevibacterium were reduced in proportion to the exposure time to CCCP. The lethal effect of CCCP to Brevibacterium was slightly enhanced by the addition of glucose into the culture. PMID- 7476557 TI - Cure of prophage in Staphylococcus aureus by furocoumarin photoadditions. AB - In a previous report curing of prophage in Staphylococcus aureus by treatment with 8-methoxypsoralen plus near-UV (365 nm) radiation was described. In the present work the use of angelicin and 4,5',8-trimethylpsoralen and repeated irradiation show that monoadducts and DNA interstrand cross-links formed by furocoumarin photoaddition are involved in the curing of the prophage responsible for the suppression of the lipolytic activity in a putative repair proficient strain of Staphylococcus aureus. PMID- 7476558 TI - Local skin reaction in mice and guinea pigs induced by a single intradermal inoculation of Fusobacterium necrophorum lipopolysaccharide. AB - Dermal responses induced by Fusobacterium necrophorum strain VPI 2891 lipopolysaccharide (LPS) were studied using mice and guinea pigs. In ddY mice, the LPS elicited inflammatory, haemorrhagic lesions and an increase in local vascular permeability 24 h postinjection. Of the mouse strains, C3H/HeJ mice were less sensitive. The LPS induced erythema and haemorrhagic responses in guinea pig skin 24 h postinoculation. These responses were dose-dependent. The intensity of dermal inflammation-inducing activity of F. necrophorum LPS was similar to that of Escherichia coli strain 055:B5 LPS, but weaker than that of Salmonella typhimurium LPS. These findings suggest that the fusobacterial LPS may play an important role in contributing to produce the initial lesions in the bacterial infection. PMID- 7476559 TI - Cell surface hydrophobicity and the net electric surface charge of group B streptococci: the role played in the micro-organism-host cell interaction. AB - The cell surface hydrophobicity, net electric surface charge and cell adhesion of six group B streptococci strains were assessed. Treatment with trypsin reduced cytoadhesion of the six strains (80340, 90356, 85147, 90222, 90186 and 88641) and induced loss of surface negative charge in the other four strains (80340, 85147, 90222 and 90186). The same treatment increased the surface hydrophobicity of three strains (90356, 90222 and 88641). Neuraminidase treatment caused a decrease in the negative surface charge of all the strains resulting in significant increases in both cytoadhesion and surface hydrophobicity of five (80340, 90356, 85147, 90222 and 88641) and four (90356, 85147, 90222 and 88641) strains, respectively. This indicates that sialic acid residues are important anionogenic groups exposed on the streptococcal cell surface. Treatment of buccal epithelial cells with N-acetyl-beta-D-glucosaminidase made them less adherent for most of the strains (80340, 85147, 90222, 90186 and 88641) assayed. PMID- 7476560 TI - N-terminal amino-acid sequence of beta-lactamase from Shigella flexneri UCSF-129. AB - A beta-lactamase (EC 3.5.2.6 penicillinase, penicillin amino beta-lactam hydrolase) was purified from Shigella flexneri USCF-129 by an efficient two-stage procedure involving chromatography in Sephadex G-75 and HPLC on a C18-reverse phase column. The homogeneity of the purified enzyme was confirmed by capillary zone electrophoresis (CZE), HPLC electrospray mass spectrometry (LC-ESMS) and amino acid sequence analyses. The highly purified enzyme was a monomeric protein with a molecular mass of 28.903 +/- 2 Da, as determined by LC-ESMS. The amino acid sequence of the first 49 N-terminal residues of this beta-lactamase revealed 100% similarity with the mature forms of the plasmid coded Escherichia coli enzymes (plasmid pBR 322 and R6K) a TEM-type beta-lactamase. PMID- 7476561 TI - Chemical microanalysis of hyphae and conidia from Arthrinium aureum. AB - Analysis of the chemical composition of hyphae, conidia and chlamydospores of Arthrinium aureum enabled all sample constituents of > 23 kD molecular weight to be detected. Sulphur, chlorine, potassium and calcium were present in the hyphae and condia of A. aureum, and sulphur, potassium and calcium in the chlamydospores. PMID- 7476562 TI - In situ mapping of urease-positive areas in porcine gastric mucosa. AB - Urease activity is a feature of gastric helicobacters, and its abundant production provides an indirect means of detecting their colonization. A method for mapping urease-positive areas directly on the gastric mucosa was developed, and 57.8% of pigs had evidence of Helicobacter colonization based on urease assay. Moreover, 89.2% of urease-positive pigs had gastritis, confirming that the known association found in man between Helicobacter and gastritis was found also in pigs. The proposed urease assay allowed detection of all urease-positive areas on the gastric mucosa, thus overcoming the biopsy sampling problems derived from the patchy distribution of helicobacters. In this way, gastric mucosa specimens from urease-positive areas could be usefully utilized for culture attempts and for microscopic examination. PMID- 7476563 TI - Apollo MEED mycology revisited and reviewed, including the Trichophyton terrestre keratinophilic growth at splashdown and 23 years after exposure to space parameters. AB - Keratinophilic Trichophyton terrestre conidia were exposed to selected parameters of space flight including 254, 280 and 300 nm UV light, full light and total darkness of space. Phenotypic isolates were grown on human hair collected from one source at years 1 and 23 after splashdown. The patterns of fungal growth on the hair, and the hair deterioration rates, were noted according to the space exposure. Growth and deterioration were consistent but slightly reduced at year 23. PMID- 7476564 TI - Biodiversity of lactic acid bacteria from food-related ecosystems. AB - The diversity of lactic acid bacteria within a variety of food-related ecosystems was assessed and the strain as well as environment specific characteristics were investigated. The strains (108 in toto) were isolated from plant material, traditional fermented foods, dried marine algae, sea food, fungi as well as spoiled foods and beverages, and all were characterized to genus level. Lactobacillus strains dominated all ecosystems and contributed to 65% of the isolates while a further 13% consisted of Lactococcus strains, 12% of Leuconostoc strains, 5% of Pediococcus strains, and 5% of unidentified strains. Plant material was the most diverse ecosystem containing representatives from each of the genera as well as the majority of dextran and tyramine producers. Those strains able to grow at 45 degrees C as well as acid tolerant strains were predominantly isolated from traditional fermented beverages while halotolerant strains occurred mainly in sea food. Two bacteriocin producers but no histamine producers were isolated. PMID- 7476565 TI - Antibiotic activity of marine algae against multi-antibiotic resistant bacteria. AB - The antibiotic activity of six species of marine algae (Rhodophyta, Phaeophyta and Chlorophyta) against multi-antibiotic resistant (MAR) bacteria was investigated. The study shows that various degrees of activity were present in 18 out of the 24 algal extracts. The highest activity was for Rhodophyta (diameter of the inhibited zone ranged from 10-22 mm), whilst the lowest was for Chlorophyta (8-12 mm). The taxonomic trends were seen between extracts, as the best solvents were found to be ether, methanol and acetone for Rhodophyta, Phaeophyta and Chlorophyta, respectively. The importance of these results on public health are discussed. PMID- 7476566 TI - Gluco-oligosaccharide production by a strain of Erwinia amylovora. AB - When grown in sorbitol-asparagine-salts medium, cells of Erwinia amylovora isolate T83 released a mixture of oligosaccharides and lipopolysaccharide in place of extracellular polysaccharide. The oligosaccharide fraction was usually readily separable from lipopolysaccharide by gel permeation chromatography, but some samples required disaggregation by preliminary treatment with EDTA. By 1H- and 13C-NMR spectroscopy, methylation analysis and mass spectrometry, the oligosaccharide fraction was shown to consist of terminal, beta-1,2- and beta 1,2,6-linked D-glucose residues in the ratio 2.2:2.6:1, indicating a branched oligosaccharide structure. The purified oligosaccharide fraction was partially resolved by gel permeation chromatography into two components containing approximately equal amounts of glucose. When grown on pear slices, isolate T83 produced normal extracellular polysaccharide. PMID- 7476567 TI - Role of lymphocyte function-associated antigen-1 on the interplay between lipid A activated monocytes and polymorphonuclear cells. AB - Previous findings provided evidence that bacterial lipopolysaccharide (LPS) activated human monocytes are able to upregulate autologous polymorphonuclear (PMN) phagocytic ability via cell-to-cell contact mechanisms mediated by membrane (m)-associated cytokines (CKs), such as tumour necrosis factor (TNF)-alpha, interleukin (IL)-1 alpha, IL-1 beta, IL-6 and IL-8. Consequently, the role of the lymphocyte function-associated antigen (LFA)-1 molecule on the monocyte (Mo)-PMN interplay was evaluated. In the first step, lipid A (LA)-stimulated Mo were pretreated with anti-recombinant human (Rhu) LFA-1 alpha monoclonal antibody (MoAb), and the enhanced phagocytic activity of PMN was abrogated. Pretreatment of unstimulated Mo with the same MoAb led to a reduction of PMN phagocytosis. In the second step, the role of m-LFA-1 on PMN was investigated with regard to Mo modulation. Anti-Rhu LFA-1 alpha MoAb was supplemented to LA-activated and unstimulated PMN, respectively, before coculturing with autologous LA-activated Mo. The addition of anti-Rhu LFA-1 alpha MoAb gave rise to a significant decrease in PMN phagocytosis regardless of PMN activation. These data suggest that, besides m-CKs, LFA-1 present on Mo and PMN might be involved in the mutual interplay between PMN and Mo. PMID- 7476568 TI - Isolation, cultivation and partial characterization of a kinetoplastid flagellate from the hindgut of the water bug, Lethocerus indicus. AB - A new species of kinetoplastid flagellates was isolated from the hindgut of the giant water bug, Lethocerus indicus, and successfully cultured in modified glycerol beef extract medium. Light and electron microscopic studies as well as growth in various culture media are described. The flagellates were in three different shapes and sizes in the hindgut of the host as well as in the culture medium. Each form of the parasite contained a kinetoplast which was located near the basal body of the flagellate. The ultrastructural features of this species exhibit great similarity to other species of Bodo. The flagellate ingested bacteria, grew in culture, and was identical with that observed in the hindgut of the bugs. The name Bodo indica has been proposed for this kinetoplastid flagellate. PMID- 7476569 TI - Salmonella exclusion in broiler chicks by the competitive action of adult gut microflora. AB - A series of experiments was carried out to investigate the efficacy of the competitive exclusion technique in the control of Salmonella in broiler chicks. Anaerobic overnight culture obtained using 10(-3) g ml-1 caecal material in VL broth medium provided total exclusion of a moderate S. enteritidis challenge (1.2 x 10(2) cells/chick). Increasing the challenge level resulted in less protection, but significant protection occurred at an excessive S. enteritidis challenge level of 1.5 x 10(6) cells/chick. The effectiveness of protection using both moderate and excessive challenges was monitored in newly hatched chicks until they were 22 days old. An improvement in growth performance in terms of higher body weight, feed consumption and feed conversion, in addition to lower mortality, was observed. The use of other control measures such as antimicrobial feed additives or lactose sugar (1.5% w/v) in the drinking water in combination with the competitive microflora treatment resulted in a better overall protection and performance. PMID- 7476570 TI - Hydrogen peroxide-induced increase in lung endothelial and epithelial permeability--effect of adenylate cyclase stimulation and phosphodiesterase inhibition. AB - Neutrophil-derived hydrogen peroxide (H2O2) is believed to play an important role in inflammatory lung injury. We investigated the influence of pharmacological agents that increase intracellular c-AMP levels on endothelial and epithelial leakage in response to intravascular H2O2 challenge in buffer-perfused rabbit lungs. Endothelial permeability was assessed by determination of the capillary filtration coefficient (Kfc) and lung weight gain. Measurement of the clearance rate of inhaled aerosolized technetium-99m-labeled diethylenetriamine pentaacetic acid ([99mTc]DTPA) from the lungs into the perfusion fluid was used as an index of alveolar epithelial permeability. Experiments were performed in the presence of acetylsalicylic acid to suppress H2O2-induced lung prostanoid generation and concomitant vasoconstriction. Under these conditions, H2O2 admixture to the perfusate (250 microM) caused a greater than eight-fold increase in Kfc values, resulting in > 30 g lung weight gain within 30 min in the absence of any significant vasopressor response. Pretreatment with the adenylate cyclase activators prostaglandin E1 (0.1 microM) and forskolin (0.1 microM), the dual phosphodiesterase type III/IV inhibitor zardaverine (10 microM) as well as combinations of these drugs all caused a nearly complete suppression of this early Kfc increase; and severe edema formation (> 30 g) was retarded to approximately 50-55 min. In addition to the microvascular leakage response, H2O2 caused a four- to five-fold increase in the [99mTc]DTPA clearance rate, starting within 15 min and culminating after approximately 35 min. Adenylate cyclase activation reduced this epithelial leakage response by approximately 30%, whereas zardaverine exerted no significant effect. We conclude that both microvascular endothelial and alveolar epithelial barrier function are severely compromised by intravascular H2O2 challenge in intact lungs. Pharmacological approaches to increase c-AMP levels, including both adenylate cyclase activation and phosphodiesterase inhibition, partially block the endothelial response and, to a lesser extent, the epithelial response. PMID- 7476571 TI - Dog model for cerebrovascular studies of the proximal-to-distal distribution of sequentially injected emboli. AB - This study was undertaken to determine whether microemboli injected in a predetermined sequence would maintain that sequence once they came to rest in brain microvessels. If so, the injection of different-colored microspheres at different times could be used to bracket-in-time emboli that are known to be released into the circulation during cardiopulmonary bypass. We injected different-colored microspheres into the arterial circulation of anesthetized dogs before and after the injection of fat emboli and before and after cardiopulmonary bypass. Coronal slices of the dog brains were embedded in celloidin, sectioned at 100 microns, and stained for alkaline phosphatase. The afferent cerebrovasculature stained dark brown against a light background, and the proximal/distal orientation of many of the arterioles could be determined by following their course within the thick sections. When different types of emboli were found in a single arteriole, they appeared in the order injected or the order of occurrence in the bypass protocol in 99.3% of the 867 such arterioles counted. Therefore, the microemboli maintained their ordered sequence with only a very small degree of mixing. Once they came to rest, there was not sufficient collateral blood flow in the brain microvessels to move them into disordered positions. This dog model should facilitate studies of the time of release of microemboli within narrower windows of time during cardiopulmonary bypass. PMID- 7476572 TI - Efficient isolation of rat aortic endothelial cells by elimination of contaminating cells with a monoclonal antibody. PMID- 7476574 TI - Glial-conditioned medium elevates alkaline phosphatase activity in cultured bovine aortic endothelial cells. PMID- 7476573 TI - A new simple method for isolation of microvascular endothelial cells avoiding both chemical and mechanical injuries. AB - Our study indicates that when small pieces of lung or muscles of chest wall are cultured, erythrocytes and leukocytes (PMNs) leave the tissues first, followed by vascular endothelial cells (ECs). Fibroblasts and other mixed cells grow after 72 hr culture. The ECs can then be isolated avoiding mechanical and chemical injuries. The lung tissue is obtained from the peripheral surface and muscles from the chest. It is then cut into pieces and cultured with DMEM containing 20% fetal bovine serum. After 60 hr culture, the tissues are discarded. The flask contains only ECs and blood cells. Blood cells can be cleared out after the cells are subcultured once or twice. The primary cells and the subcultured cells cultured on gelatinized culture dish give the capillary-like structure. Cells cultured on untreated dishes have regular cobblestone morphology and junctional contacts. The isolated cells were not mesothelial cells because the cells did not react to antibody against cytokeratin 18, while mesothelial cells reacted strongly to the antibody. The cells can be isolated from the lung tissue without pleura. The primary microvascular ECs are also cultured on microcarriers (cytodex 3). Because both mechanical and proteolytic injuries are avoided, the cells may be more similar to cells in the in vivo state. There are no significant differences in PMN-endothelium adherence and monolayer responses to second messengers, platelet activating factor, and phospholipase A2 when pulmonary and muscular microvascular endothelial cells are compared. PMID- 7476575 TI - Effect of basic fibroblast growth factor on microvascular regeneration from gastric ulcerative lesion-increased binding site of bFGF after CS23 treatment. PMID- 7476576 TI - Effects of monoionic and nonionic radiographic contrast media on cutaneous microcirculation in patients with peripheral arterial occlusive disease. AB - The purpose of the study was to compare the in vivo effects of two low-osmolar radiographic contrast media, a monoionic (sodium/-meglumine ioxaglate, Hexabrix) and a non-ionic one (iopromide, Ultravist 300) on the cutaneous microcirculation in patients with peripheral arterial occlusive disease in a prospective randomised double-blind study. In 20 patients with peripheral arterial occlusive disease scheduled for percutaneous transluminal angioplasty skin microcirculation was assessed by laser Doppler fluxmetry and transcutaneous oxygen tension measurements at the foot dorsum for continuous recordings of the cutaneous microvascular perfusion and skin oxygenation. Before angioplasty 10 ml nondiluted contrast medium was injected intraarterially and the acute effect on transcutaneous oxygen tension and laser Doppler flux was registered. Mean laser Doppler flux and oxygen tension were evaluated initially, after arterial puncture and after contrast medium injection. In addition, flux motion was analysed with respect to frequency and amplitudes changes. No significant changes in laser Doppler flux and fluxmotion patterns were found between both groups after contrast medium injection. However, mean transcutaneous oxygen tension of the patients receiving monoionic ioxaglate dropped significantly (P = 0.01). The differences of both contrast media on red blood cell aggregation shown in vitro do not correlate to intravital microcirculatory measurements in patients with arterial occlusive disease of the lower limb. Microvascular skin blood flow and flux motion at the foot in these patients do not change significantly after intraarterial injection of ioxaglate and iopromide. However, oxygen supply of the skin is transient impaired after interarterial injection of the anionic contrast agent ioxaglate, which was not associated with clinical symptoms. PMID- 7476577 TI - Identification of a melanocortin receptor expressed by murine brain microvascular endothelial cells in culture. AB - Microvascular endothelial cells were isolated from the brains of C57 mice and cultured in selective growth media. The isolation and culture techniques employed in this study minimised the contamination by nonendothelial cells such as astrocytes, pericytes, and smooth muscle cells. Microvascular endothelial cells examined using phase contrast light microscopy grew as small colonies of spindle shaped cells which merged together to form typical contact-inhibited monolayers. The endothelial origin of these cells was determined using several established characterisation techniques. Preliminary receptor binding studies at 4 degrees using [125I-Tyr2, Nle4, D-Phe7]alpha-melanocyte-stimulating hormone ([125I-Tyr2, Nle4, D-Phe7]alpha-MSH) suggested the possibility that melanocortin receptors were present on the surface of brain microvascular endothelial cells. Subsequent binding isotherms confirmed that a small population of high-affinity melanocortin receptors was expressed. The existence of a specific binding site for alpha-MSH was confirmed by photoaffinity labeling with the 4-(1-azi-2,2,2, trifluoroethyl)benzoic acid (ATB) derivative, [125I-Tyr2, Nle4, D-Phe7, (ATB) Lys11] alpha-MSH. SDS-PAGE analysis identified the presence of a specific band with a molecular mass of approximately 45 kDa, which was consistent with previous data on melanoma melanocortin receptors, and represented a ligand-receptor complex. This study suggests that a receptor for alpha-MSH is expressed on the extracellular surface of murine brain microvascular endothelial cells; however, the physiological role of this receptor is as yet unknown. PMID- 7476578 TI - Interaction of activated natural killer cells with normal and tumor vessels in cranial windows in mice. AB - A mammary carcinoma, MCa IV, was grown in syngeneic C3H mice in a cranial window preparation which permitted the in vivo observation of the growth and microcirculation of the tumors. Fluorescently labeled activated natural killer (A NK) cells were injected into the external carotid artery and their interactions with normal and tumor vessels were quantified by video microscopy. Cells which entered the tumor vessels adhered heterogeneously to these vessels, regardless of vessel size or blood flow rates and bound with an efficiency ranging from 0 to 82% of the incoming cell flux. Normal brain tissue showed significantly fewer binding cells per microscopic field (9 +/- 5 vs 85 +/- 27 cells/1.3 mm2) and the few cells which were retained by the normal tissue were highly deformed, suggesting mechanical rather than adhesive entrapment. These studies indicate that A-NK cells bind in high numbers to segments of the vessels of mammary tumors growing in an intracranial site when administered through an arterial route; however, some tumor vessels may escape recognition by these cells. These findings suggest that A-NK cells may be used as carriers of genes for anti-cancer agents. PMID- 7476579 TI - Microendoscopy of the synovial vasculature in the rabbit knee joint. AB - The functional morphology of the synovial vasculature in the rabbit knee joint was investigated using our technique of microendoscopy, which involves an in vivo microscopy via a 1.9-mm endoscope. In the vessels of the fibrous tissue of the joint capsule it was found that after intraarticular administration of epinephrine the blood flow and the vascular diameters decrease for 3-5 min. In the areolar part of the synovial tissue, an external elevation of the intraarticular pressure causes collapse of the capillaries at 10-15 mm Hg, whereas the collapse pressure of the arterioles is 30-40 mm Hg. Intraarticular application of Neuropeptide Y (NPY) leads to a marked vasoconstriction including the arteriovenous anastomoses, whereas after epinephrine the arteriovenous anastomoses remain patent or dilate during the arteriolar constriction phase. It is concluded that the technique of microendoscopy allows a valuable in vivo investigation of the synovial microvasculature. PMID- 7476580 TI - Polyarginine and eosinophil-derived major basic protein increase microvascular permeability independently of histamine or nitric oxide release. AB - We tested the hypothesis that cationic peptides (polyarginine; poly-argn) and eosinophil-derived major basic protein (MaBP) increase permeability by stimulating the release of histamine and/or nitric oxide. We used intravital microscopy, clearance, and integrated optical intensity (IOI), using FITC-dextran 150 (FITC-dx 150) as a tracer, to evaluate changes in microvascular permeability in the hamster cheek pouch. Poly-argn at 1 microM (topical) increased the clearance of FITC-dx 150 from 610 to 3240 nl/60 min/g. In contrast, 1 microM polyglutamic acid (poly-glun; and anionic peptide) did not affect the clearance of FITC-dx 150 (605 nl/60 min/g). At 0.5 and 1.0 nM, poly-argn increased the clearance of FITC-dx 150 from 610 to 1722 and 2396 nl/60 min/g (P < 0.05). Similarly, 0.5 nM MaBP increased clearance from 591 +/- 38 to 1967 +/- 168 nl/60 min/g. L-NAME at 10(-4) M did not prevent the 0.5 nM MaBP-induced elevation in clearance (1784 +/- 350). Poly-argn at 1 nM increased net mean IOI by 21.5 +/- 7.2 units. This elevation was not inhibited by topical 10(-4) M L-NMMA (27.5 +/- 8.4). Using 0.5 nM poly-argn as agonist, we assayed suffusate samples for their histamine concentration using a competitive enzyme immunoassay and found no detectable histamine. Pyrilamine, an H1 antagonist, did not inhibit the 0.5 nM poly-argn-induced elevation in clearance of FITC-dx 150. We conclude that (1) cationic peptides and MaBP increase microvascular permeability and (2) the increase in microvascular permeability produced by low concentrations of poly argn and by MaBP is independent of the release of histamine and does not require nitric oxide. PMID- 7476581 TI - Attenuation of microvascular permeability dysfunction in postischemic striated muscle by hydroxyethyl starch. AB - We examined the effect of hydroxyethyl starch macromolecule (HES-Pz) pretreatment on microvascular transport of macromolecules in ischemia-reperfusion injury. The rat cremaster was splayed, placed in a Lucite intravital chamber, and suffused with bicarbonate buffer. The clearance of fluorescein isothiocyanate dextran 150 (FITC-Dx 150) was measured as an index of microvascular transport. After determination of baseline data, the muscle was made ischemic for 4 hr by clamping the vascular pedicle and subsequently reperfused for 2 hr. In control animals not subjected to ischemia, clearance of FITC-Dx 150 remained constant throughout the experimental 7-hr period. In saline-treated animals, ischemia-reperfusion increased the clearance of FITC-Dx 150 from 1.8 +/- 0.3 to 9.7 +/- 1.0 microliters/15 min/g by the end of the reperfusion period. Pretreatment with HES Pz, at a concentration of 6% in a volume of saline equivalent to 10% of blood volume, significantly attenuated the microvascular dysfunction produced by ischemia-reperfusion. The mean ratio of postischemic to baseline clearance of FITC-Dx 150 was 1.28 +/- 0.07 (mean +/- SE) for samples taken from the 30th to the 120th min of reperfusion at 15 intervals. Our data support a beneficial effect of HES-Pz on microvascular transport of macromolecules. The role of leukocyte-endothelium adhesion as an underlying mechanism explaining these results was studied by evaluating the effect of HES-Pz on the ability of thrombin stimulated human umbilical vein endothelial cells (HUVECs) to bind neutrophils. These experiments demonstrated that thrombin-treated HUVECS bound 229% more indium-111-labeled neutrophils than did similarly stimulated HUVECS treated with HES-Pz (P < 0.05).(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7476582 TI - Vasoactive agonists do not change the caliber of retinal capillaries of the rat. AB - The pericytes of the retinal capillaries are considered to be contractile cells, based on indirect evidence. We attempted to detect pericyte contractility directly using a morphometric method following intravitreal injection of vasoconstricting (endothelin-1, norepinephrine) and vasodilating (forskolin, prostaglandin E1) agonists in rats. The technique involved measuring changes in the ratio of the (lumenal perimeter x mean lumenal radius) to the lumenal area (rP/A ratio, which we defined as "caliber"). Following intravitreal injection of endothelin-1 or norepinephrine there is an ophthalmoscopically visible constriction of the larger retinal arterioles. This constriction is also clearly demonstrable morphometrically (P = 0.001 for endothelin-1 and P = 0.047 for norepinephrine). However, neither for the vasoconstricting nor for the vasodilating agonists is there evidence of a significant change of the caliber of the capillaries. At least in the retina of the rat, therefore, we have no evidence that the pericytes perform a contractile function that alters flow in the capillaries. PMID- 7476583 TI - Spontaneous flow waves detected by laser Doppler in human skin. AB - The contribution of spontaneous variations in sympathetic nervous activity and perfusion pressure to the laser Doppler flux signal in human skin was studied in nine healthy subjects. Simultaneous recordings were made of laser Doppler flux, mean blood pressure, and blood flow in the radial artery. In the skin of the palm and sole, there was a significant and strong correlation between fluctuations in flux and radial artery velocity, which indicated a high degree of neural control of microcirculatory blood flow. This correlation decreased progressively toward the trunk and face. In the skin of the nose and at scattered sites on the face, trunk, and extremities, there was correlation between flux and mean blood pressure that indicated a predominantly "passive" vascular bed. At other sites, multivariate regression analysis revealed contributions of both blood pressure and sympathetic activity. In skin on the ear and forehead, a characteristic pattern of regular oscillations in flux, uncorrelated with blood pressure and sympathetic activity, was demonstrated. The laser Doppler signal from different skin sites thus contains varying contributions from variations in perfusion pressure, sympathetic nervous activity, and local, myogenic arteriolar vasomotion. PMID- 7476584 TI - Hypothetical substrates of ideation compared in terms of mechanical and quasimechanical coherence of behavior. AB - Relationships between physiologic models and ideas in general require axioms for their expression. Axioms proposing that a given human's ideation is borne in trajectories of response owing their coherence to mechanical arrangement limit neurologic therapy to the arbitrary deflection of patients toward target behaviors. However, a patient's physiologic acts of describing environment, if coherent yet mechanically disjoint at their origins, could be proposed as ideationally-generated. Neurologic therapy could then be the aim to provide for their optimal use. PMID- 7476585 TI - A novel role for surfactant in the lung with implications for the sudden infant death syndrome. AB - Feedback from mechanoreceptors in the lungs to the brainstem has long been known to be vital for control of rhythmogenesis during normal breathing. Action potentials transmitted via the vagi are shown to display an irreversible relationship with respect to lung area (A), i.e. hysteresis, which closely resembles the hysteresis in surface tension (gamma) versus A for normal surfactant which coats that surface. Since lung recoil is largely determined by surface tension for resting tidal volumes, this will dominate the stretch of mechanoreceptors. Hence, it is postulated that, through the Hering-Breuer reflex, surfactant largely determines afferent neural feedback, explaining the above similarity in clockwise hysteresis loops. Thus the ability of normal surfactant to impart normal clockwise gamma: A hysteresis is seen as a desirable property enabling the brainstem to differentiate between the inspiratory and expiratory phases of the breathing cycle at the same lung volume. It is further hypothesized that the very abnormal surfactant found recently in some infants with prolonged expiratory apnoea by displaying anti-clockwise gamma: A loops would render afferent neural feedback to the brainstem highly confusing and could cause prolonged expiratory apnoea. This concept is discussed as a possible cause of sleep apnoea, recurrent cyanotic episodes and the Sudden Infant Death Syndrome (SIDS). PMID- 7476586 TI - Extracellular nucleic acids as possible messengers in regulatory information transfer from immune system to non-lymphoid tissues. AB - The goal of this review is to connect the information on the regulation of growth and differentiation of non-lymphoid tissues by the immune system and the release/uptake of nucleic acids by living cells (especially by lymphocytes). Nucleic acids are suggested to be the messengers in regulatory information transfer from the immune system to other cells. PMID- 7476587 TI - Involvement of endogenous benzodiazepine receptor ligands in brain disorders: therapeutic potential for benzodiazepine antagonists? AB - Many therapeutic effects of benzodiazepines are mediated by neuronal high affinity binding sites, i.e. benzodiazepine receptors (BR), located on GABAA receptors. Recently, endogenous BR ligands have partially been identified which, as agonists, either increase or, as inverse agonists, decrease GABAergic inhibition in the brain. BR antagonists, previously described as intrinsically inactive, induce effects in animals and humans under particular circumstances emphasizing a functional relevance of endogenous BR ligands. Several brain disorders, e.g. anxiety, insomnia, epilepsy, spasticity, alcoholism, coma, dementia, may be associated with a disequilibrium of opposing endogenous BR ligands changing the excitability of neurons implicated in aforementioned diseases. It is proposed that, depending on the relative role endogenous BR ligands play in the pathophysiology of these disorders, BR antagonists might demonstrate a variable efficacy in improving their symptomatology. In fact, such therapy would restore the homeostatic balance among various endogenous BR ligands being disturbed during an illness. PMID- 7476588 TI - The Hippocratic Oath: a code for physicians, not a Pythagorean manifesto. AB - The Hippocratic Oath is to be considered a code of conduct for all physicians and not a Pythagorean manifesto, in spite of the view of Edelstein. In fact, it can be shown that the prohibitions and requirements on which the Pythagorean hypothesis rests (the prohibition against helping suicide, inducing abortion, performing surgery, and having sex with patients or with members of their household and the rules of confidentiality and collegiality) do not necessarily link the Hippocratic Oath to the Pythagoreans. Edelstein affirms that only the Pythagoreans condemned suicide, whereas it can be shown that Plato, Aristotle, the Epicureans, and several authors in antiquity opposed it. Similarly, induced abortion was by no means universally accepted in antiquity. Soranus, for example, clearly states that many physicians opposed it in all cases. The passage of the oath concerning surgery can be shown to refer only to lithotomy (as others have underlined). As for sexual relations with patients or members of their household (male or female), the existence of laws against promiscuity (homo- or heterosexual), and other evidence, indicates that it was usually condemned. Finally, confidentiality and collegiality were virtues that the Pythagoreans were not alone in upholding. In addition, many of the principles upheld by the Oath are found in other documents unrelated to the Pythagoreans. PMID- 7476589 TI - Parametric stimulation of hemopoietic stem cells in normals and in certain leukemias. AB - The paper discusses certain distributive and clustering modes of stimulus P and of its receptors. When combined with topologic features peculiar to sources and stem cells, these parameters account not only for the cycling variations typical of normal stem cells but also for seemingly paradoxical shifts in proliferative activity of cells grafted into patients with chronic myeloid leukemias. PMID- 7476590 TI - Correlation between spin lattice relaxation time and radiation sensitivity in human tissues. AB - The water proton spin lattice relaxation time of normal and tumorous tissues of human and radiosensitivity yield a linear relationship. PMID- 7476591 TI - Local transfer of hormones between blood vessels within the adrenal gland may explain the functional interaction between the adrenal cortex and medulla. AB - The adrenal cortex has a local stimulatory action on synthesis and secretion of adrenaline by the adrenal medulla. This interaction may be based on nervous transmission and/or on glucocorticoids secreted trom the cortex either exerting paracrine effects or reaching the medulla through the local vascular system. The existence of a dual capillary network, a portal system, now seems unlikely. However, a new concept is postulated: the high demand for glucocorticoids in the medulla may be met by local transfer to blood vessels within the adrenal gland, particularly the medullary arteries. PMID- 7476592 TI - Hair growth and the fluid factor. AB - It is accepted that the detailed mechanisms of changes in human hair growth patterns are poorly understood. It is this uncertainty that has encouraged many charlatans to operate in this area. From my perspective as an engineer, however, there is a simple mechanism that makes sense of these changes. I suggest this mechanism is closely connected with the evolution and function of hair. The fact that the presence of this mechanism can be demonstrated in the male pattern baldness scenario, raises a number of serious questions relating to fundamental physiology and the mechanisms of some serious diseases. As an amateur, it is difficult for me to gain access to up-to-date research data, so my references are derived from textbooks. I would therefore welcome comments from professionals who are involved in the areas indicated in this paper. PMID- 7476593 TI - Morpho-densitometry of bone using a trigonometric function. AB - When long bone is considered to be a tube-like structure filled up with a homogenous material, single-directional densitometry of it separately shows cortical and medullary bone mass using a trigonometric function at any given section. Both bone masses can be differentiated from each other using the values of peak bone density and central bone density. This densitometry is based on the two-dimensional structure of long bone. This principle can be universally applied to skeletal disorders including osteoporosis, metabolic bone disorders, and bone tumors in which the mineral content is altered. Dynamic changes in calcium metabolism can be analyzed serially without any invasion at low cost. PMID- 7476595 TI - Pantothenic acid deficiency as the pathogenesis of acne vulgaris. AB - For years, the pathogenesis of acne vulgaris has been known to be strongly influenced by hormonal factors. However, the exact role of and the interrelationship among the various hormones in question have not been well elucidated. Here, I wish to suggest a radically different theory for its pathogenesis and relate its basic pathology to a deficiency in pantothenic acid, a vitamin hitherto not known to cause any deficiency syndrome in humans. Hence, the effect of hormonal factors in this disease entity becomes secondary to that of the availability of pantothenic acid. A complete cure of this condition is effected by a very liberal replacement therapy with the vitamin. PMID- 7476596 TI - Plant bacterial spores, active systemically as a separate entity, play a significant role in human illnesses such as cancer, granulomas, AIDS, and milky white abdominal ascites that currently defies recognition. AB - There is currently an unrecognized chapter in medical illnesses, occurring in living human beings, that defies recognition in explaining the diseases' origin and growth, and failing accurately to account for the pathophysiology involved. It is pertinent, therefore, to alert medical science, based upon facts as uncovered by my research studies (1-5), that plant bacterial spores can exist as a separate entity systemically. Being totally devoid of the presence of their adult origin, they can survive with a degree of viability. Ultimately, they may become embedded within, or nearby, a network of cells, consisting of the reticuloendothelial, squamous, or epithelial group. With an ensuing adequate but compatible circulating flow of blood by the host to the specific tissue site as muscles, bone, or bone marrow, it can thus lead to the various pathophysiological changes and illnesses that currently defy an accountable recognition. PMID- 7476594 TI - HIV and the cortisol connection: a feasible concept of the process of AIDS. AB - Recent evidence suggests that HIV infection and the clinical and laboratory manifestations of acquired immunodeficiency syndrome (AIDS) are a result of the genetic influence of the virus on cellular adrenocorticotrophic hormone (ACTH) and cortisol metabolism. Recent genetic studies substantiate this view with the observation that the HIV-1 genome is linked to glucocorticoid inducibility and to glucocorticoid receptor binding, and may explain the strong ability of cortisol to enhance HIV replication. Adrenocortical hyperactivity observed in HIV-infected individuals has been found to be independent of the hypothalamic-pituitary axis, and is apparently a result of increased ACTH production by HIV. It is proposed that the HIV-induced cortisol excess is the foundation of the immunosuppression seen in AIDS, and is the basis for alternative avenues of treatment, including the use of ascorbic acid. PMID- 7476597 TI - Working memory networks and the origin of language areas in the human brain. AB - Temporoparietal-prefrontal working memory networks are proposed as fundamental in the evolutionary origin of the language regions. Having a primordial capacity to name objects or situations, primitive hominids may have strongly benefited from the possibility to recall past events from memory, in order to refer to them through vocal communication. Working memory cortical networks are related to these types of tasks, and are arranged quite similarly to the language networks in the brain. It is possible that the language areas and their connections arose as a local specialization of these large-scale cortical networks, that developed as neural strategies to recall past events to be shared in community. The origin of syntax may have taken place after these networks were sufficiently stabilized, and (at least originally) may have been related to aspects of vocal motor control, involving the progressive differentiation of the anterior language areas and their connections. PMID- 7476598 TI - Chronic intestinal candidiasis as a possible etiological factor in the chronic fatigue syndrome. AB - The chronic candidiasis syndrome, also known as the Candida-related complex, putatively caused by the overgrowth of Candida albicans in the gastrointestinal tract and secondarily in the genital organs, is briefly described. Patients with this disorder have many of the same symptoms as those with the chronic fatigue syndrome, except for the recurrent flu-like symptoms of the latter disorder. The positive response of a large number of patients with the chronic fatigue syndrome (CFS) to an oral antifungal agent and a diet for intestinal candidiasis has been described by another clinician. There is evidence that Candida albicans infection of the mucous membranes depresses T cell and natural killer (NK) cell function. Similar abnormalities of immune function are found in the CFS. The function of cytotoxic T cells, T helper cells, and NK cells is important in preventing reactivation of infections from Epstein-Barr virus, cytomegalovirus, and other herpesviruses. Reactivation of one or more of these viruses could lead to the expression of the flu-like symptoms in the CFS. Yet the immune dysfunction found in this disorder has been considered the primary underlying causal factor. It is proposed that chronic intestinal candidiasis may be an agent which leads to immune depression in many CFS patients and therefore that it could be a causal factor in CFS. PMID- 7476599 TI - Is bronchial asthma a pancreatic disease? AB - Bronchial asthma and diabetes mellitus seldom occur in the same patient. The exact mechanism of this mutual exclusion is still unknown and its elucidation can make clear the physiopathology of both diseases. Clinical and experimental evidences suggest that insulin is a proinflammatory hormone and glucagon an antiinflammatory and a bronchodilator one. We hypothesize that the relationship between plasma insulin and glucagon may play an important role in bronchial asthma. PMID- 7476600 TI - An intangible energy in the functioning biosystem. I: A search for its fate and a proposed method of delivery. AB - This paper treats evidence for an increasing recognition of a force with intangible properties in biosystems with special reference to its ability to transport electrons at near 0 degree Kelvin. It is implied that such force, whose demonstration at these temperatures is an experimental contrivance to emphasise its strictly quantal status, can be operative at room temperature. A discussion is entered as to a mode for delivery of intangible energy from demonstrable non local origins, locally to the cell to provide for structure and function. Extensive use is made of theory of the structure of the photon from de Broglie and others to accommodate a co-existence of real (electromagnetic) and quantal (intangible) fields within the photon and to discuss how such co-existence may have been copied or otherwise made manifest in a macroscopic structure such as the cell. Cell function is then viewed as concurrent real and intangible effects following stimuli derived from perturbation of the real or electromagnetic component. PMID- 7476601 TI - An intangible energy in the functioning biosystem. II: Useful parallels with circuit theory and with non-linear optics. AB - The argument is developed that a structure and function already exists in selected inanimate systems for an intangible energy dissipating these systems and that, in so doing, this energy exhibits certain properties, readily recognised in the functioning biosystem. The central thesis is that, during dissipation, the structure of the biosystem affords opportunity for an enhanced display of these properties, so that this structure can be rationally recognised as obligatory in the transition, inanimate to animate matter. The systems chosen are those of reactance in linear circuit theory of electronics, and some recent developments in non-linear optics, both of which rely on imaginary or quantal force to display observable effects. Discussion occurs on the fashion which the development of a statistical formalism as a basis for the study of squeezed states of light in these non-linear systems, has, at the same time, overcome a long standing veto on the practical use of quantal energy associated with the Uncertainty Principle of Heisenberg. These ideas are used to vindicate the suggestion that a theoretical basis is presently available for an engineering type approach, toward an intangible force as it exists in the biosystem. The origins and properties of such a force continue to be considered by many as immersed in mysticism. PMID- 7476602 TI - Apoptosis and HIV neuropathogenesis. AB - A consequence of HIV infection may be neurological dysfunction secondary to the presence of virus in the central nervous system (CNS). The CNS tropism of HIV and the mechanisms that govern its dissemination are not well defined. One view is that HIV enters the brain through the diapedesis of infected monocytes from blood into the perivascular space. HIV may then spread to susceptible cells throughout parenchyma. An alternate hypothesis is presented, which suggests that T lymphocyte apoptosis may also participate in HIV entry and dissemination in the brain. This is based on the following observations: 1) T lymphocyte apoptosis may be a CNS-specific mechanisms to control inflammation, 2) the most common circulating reservoir of HIV is the T lymphocyte, 3) uninfected macrophages recruited to phagocytize HIV infected apoptotic T lymphocytes in vitro can become productively infected, and 4) the predominant form of HIV in the CNS is unintegrated. Aberrantly high levels of apoptosis in HIV infected lymphocytes within the CNS and subsequent recruitment and infection of macrophages and microglia may be a novel component of HIV neuropathogenesis. PMID- 7476603 TI - Serum fatty acid/albumin molar ratio and the risk of diseases. AB - Free fatty acids (FFA) in blood are carried by serum albumin. A hypothesis is offered that conditions giving a high molar ratio of FFA to albumin may lead to dysfunction of the cells which are directly exposed to the high FFA/albumin ratio, i.e. the red and white blood cells, and endothelial cells. The hypothesis is supported by observations of (1) hemolytic effect of FFA, and protection by albumin in vitro, (2) inhibition of white blood cells by FFA, (3) increased FFA/albumin ratio and erythrocyte susceptibility to hemolysis in pre-eclampsia, (4) increased incidence of eclampsia in undernutrition, (5) the paradox at famine suppresses and refeeding activates malaria, and (6) an inverse relationship between serum albumin level and mortality. PMID- 7476604 TI - Snakes and ladders: getting patients into public hospitals. PMID- 7476605 TI - Cystic fibrosis: towards the 21st century. PMID- 7476606 TI - Lupus: advances and remaining challenges. PMID- 7476607 TI - Polio vaccination and polio eradication. PMID- 7476608 TI - Difficulties in admitting patients to metropolitan public hospitals: the view of medical practitioners. AB - OBJECTIVE: To investigate difficulties in public hospital admissions from the perspective of referring medical practitioners. METHODS: Referring surgeons, physicians and general practitioners (GPs) were randomly selected from three metropolitan (Sydney) health areas and formed into focus groups. Thematic analyses were undertaken of the groups' discussion transcripts to explore the problems of securing patient admissions, practitioners' perceptions of the reasons for these problems and the strategies they used to overcome them. RESULTS: Surgeons in two health areas reported major constraints affecting patient admissions. Physicians experienced fewer difficulties as this group was able to utilise outpatient services but was not always satisfied with such arrangements. GPs reported difficulties in communicating with full-time hospital medical staff. All practitioners felt that limited bed availability, restricted operating times, redefinition of many conditions as "non-urgent" and hospital budgetary constraints resulted in long waiting times for admission to public hospitals. This has led to an increasing move to use private hospitals for elective admissions, and to investigate patients in the community. CONCLUSIONS: Medical practitioners experienced difficulty in having their patients admitted to metropolitan public hospitals for both elective and non-elective procedures, and this was perceived to result in clinical and personal costs to patients. PMID- 7476609 TI - Are health undertakings effective in the follow-up of migrants for tuberculosis? AB - OBJECTIVE: To determine the rate of compliance of migrants issued with a health undertaking for tuberculosis requiring them to report within one month after arrival in Australia. DESIGN: Compliance was monitored prospectively between 1 November 1992 and 31 October 1993. SETTING: The Australian Government Health Service in Melbourne. PARTICIPANTS: 1660 migrants with a health undertaking for tuberculosis. MAIN OUTCOME MEASURE: Compliance with the undertaking (making initial contact and presenting for examination). RESULTS: The overall compliance rate was 58%, but 89% of those who made contact complied with their undertakings. There were significant differences in the compliance rates according to country of application (P < 0.001). It was estimated that about 150 people a year who require treatment for tuberculosis could be failing to comply with a health undertaking, eight of whom may require a full course of antituberculous chemotherapy. CONCLUSIONS: The administration of the current system of health undertakings for tuberculosis for migrants needs to be strengthened to increase compliance. PMID- 7476610 TI - Changing epidemiology of methicillin-resistant Staphylococcus aureus in Western Australia. AB - OBJECTIVE: To assess the epidemiology of methicillin-resistant Staphylococcus aureus (MRSA) in Western Australia. DESIGN: Retrospective review of statutory notification data. SETTING: Western Australia (WA), 1993. OUTCOME MEASURES: Notification rates, antibiotic resistance patterns and classification of isolates as imported or WA MRSA strains on the basis of antibiotic susceptibility. RESULTS: There were 204 notifications of MRSA, 78% of which were classified as WA MRSA. Three outbreaks of MRSA infection and colonisation occurred in separate WA hospitals. Notification rates per 100,000 were highest in the rural regions: the Kimberley (86.32), Goldfields (62.47), Mid West (37.21) and Pilbara (27.38) regions; and lowest in the metropolitan regions (5.52). All MRSA isolates were susceptible to vancomycin. Most imported strains were susceptible to amikacin, bacitracin, chloramphenicol, framycetin, fusidic acid and novobiocin, but only 23% to gentamicin. WA MRSA strains remained predominantly susceptible to all antibiotics tested, except beta-lactams, erythromycin and tetracycline, but a few strains resistant to rifampicin (1%) and fusidic acid (3%) appeared in the second half of 1993. CONCLUSIONS: The epidemiology of MRSA in WA is changing rapidly, with increases in both the numbers of notifications and the proportion from country regions. A new strain of MRSA (WA MRSA) that is less resistant to antibiotics than imported MRSA has emerged and is threatening the State's success in preventing establishment of MRSA in its hospitals. PMID- 7476611 TI - Allergy to laboratory animals among animal handlers. AB - OBJECTIVE: To determine the prevalence among laboratory animal handlers of allergy to laboratory animals and of asthma and the factors associated with their development. DESIGN: A cross-sectional survey. SETTING: Teaching and research institutions in Sydney, between January 1989 and December 1992. PARTICIPANTS: Laboratory animal handlers (teaching and research staff, animal house workers and animal husbandry students and teachers). MAIN OUTCOME MEASURES: Duration of exposure to laboratory animals, allergic symptoms on contact, skin reactivity to laboratory and domestic animal allergens and evidence of current asthma. RESULTS: 228 subjects were surveyed. Allergy symptoms occurred in 73 (56%) of the subjects exposed to laboratory animals for three months or more. This group also had significantly higher prevalences of skin reactivity to laboratory animal allergens (62%) and bronchial hyperresponsiveness (21%) than those with shorter exposure (14% and 8%, respectively). Atopic subjects exposed to laboratory animals (particularly those sensitised to domestic animals) and animal attendants (with a high intensity of exposure to laboratory animals) had significantly higher frequencies of skin reactivity to laboratory animals and asthma than other subjects (77% and 30%, respectively, among exposed atopic subjects and 84% and 33%, respectively, among animal attendants). CONCLUSIONS: Allergy to laboratory animals is an occupational hazard among laboratory animal handlers, especially for those who are atopic and sensitised to domestic animals, and may lead to the development of asthma. Screening for atopy and skin reactivity to laboratory animals before and during employment would enable those at risk to take precautions. PMID- 7476612 TI - A survey of women seeking termination of pregnancy in New South Wales. AB - OBJECTIVES: To identify characteristics of women seeking termination of pregnancy, their reasons for having a termination, and whether they intended to submit a Medicare claim for the procedure. DESIGN: Survey by self-administered questionnaire. SUBJECTS AND SETTING: All women who attended 11 private clinics in New South Wales (NSW) over a six-week period in 1992. RESULTS: 2249 completed questionnaires were received (94% response rate). Terminations were frequent at all ages, but women in their 20s accounted for more than half the terminations (54%) and teenagers accounted for only 17%. Respondents were generally well educated and from various ethnic and religious backgrounds; 56% were single and 41% had had one or more previous terminations. Fifteen per cent of respondents could not or did not intend to make a Medicare claim. The most frequently cited factors contributing to the decision to seek a termination were concerns about finances, change in lifestyle, single parenthood and being too young. CONCLUSIONS: Private clinics, where most terminations are performed, clearly meet a major demand, but it is likely that terminations in NSW as recorded by Health Insurance Commission claims are under-reported by at least 15%. PMID- 7476613 TI - Vaccine-associated paralytic poliomyelitis. PMID- 7476614 TI - Serum HDL cholesterol: should it be routinely measured? AB - HDL cholesterol is only one of many risk factors which can be used to predict coronary disease events, and the relationship between the benefits of lipid lowering strategies and changes in HDL levels is, at present quite unclear. PMID- 7476615 TI - Hospital-acquired tuberculosis. AB - The resurgence of tuberculosis and the emergence of drug-resistant strains, coupled with a growing number of immunocompromised patients and a high proportion of susceptible health care workers, have increased our awareness of the possibility of hospital-acquired tuberculosis. Infection control guidelines which aim to prevent dissemination and inhalation of infectious particles include early diagnosis and isolation of infectious patients, particular care during procedures likely to increase the density of the organism in the environment, and regular surveillance of hospital staff. PMID- 7476616 TI - The National Breast Cancer Centre. PMID- 7476617 TI - Towards a higher standard of breast cancer care. PMID- 7476618 TI - Risk factors and preventive strategies for breast cancer. AB - Several risk factors for breast cancer are readily identifiable, but most are not preventable. Hence the importance of screening, as early diagnosis and treatment offer the best hope of saving lives. PMID- 7476620 TI - Vaccination and SIDS: information from the South Australian SIDS Database. PMID- 7476619 TI - The euthanasia controversy. Decision-making in extreme circumstances. PMID- 7476621 TI - Does rheumatoid arthritis exist in the indigenous Australian aboriginal population? PMID- 7476622 TI - Malnutrition among rural Aboriginal children in the Top End of the Northern Territory. PMID- 7476623 TI - Extending the indications of registered drugs: rectal diazepam in epilepsy. PMID- 7476624 TI - Amoxycillin-clavulanic acid prescribing remains high in the elderly despite warnings of drug-associated jaundice. PMID- 7476625 TI - Schistosomiasis: beware delayed seroconversion. PMID- 7476626 TI - Lead mobilisation during pregnancy. PMID- 7476628 TI - Asymptomatic carotid stenosis: how should it be managed? PMID- 7476627 TI - Has informed consent become a legal nightmare? PMID- 7476629 TI - Excessive wax in the ears. PMID- 7476630 TI - The need for limits in medicine. PMID- 7476631 TI - Politics and publishing: the Quality in Australian Health Care Study. PMID- 7476632 TI - Men and mental health: now on the health agenda. PMID- 7476633 TI - The centenary of x-rays: celebrating the past and anticipating the future. PMID- 7476634 TI - The Quality in Australian Health Care Study. AB - A review of the medical records of over 14,000 admissions to 28 hospitals in New South Wales and South Australia revealed that 16.6% of these admissions were associated with an "adverse event", which resulted in disability or a longer hospital stay for the patient and was caused by health care management; 51% of the adverse events were considered preventable. In 77.1% the disability had resolved within 12 months, but in 13.7% the disability was permanent and in 4.9% the patient died. PMID- 7476635 TI - How safe are Australian hospitals? PMID- 7476636 TI - Medical injuries: international perspectives. PMID- 7476637 TI - The Australian Council on Healthcare Standards care Evaluation Program. AB - With the assistance of the medical colleges, the Australian Council on Healthcare Standards (ACHS), through its Care Evaluation Program, has established clinical performance measures which will assist both internal and external review of care and enable hospitals to compare their quality of patient care with that of other hospitals. PMID- 7476638 TI - Serotonin syndrome--a potentially fatal complication of antidepressant therapy. PMID- 7476639 TI - Hormone replacement therapy and breast cancer: what are the facts? AB - Selective quotation in the popular press of studies showing a potential association between hormone replacement therapy (HRT) and breast cancer has fuelled women's concern. However, the increase in detected breast cancer among long term HRT users that was seen in some studies is compatible with surveillance bias and confounding caused by increased risk factors among HRT users (e.g., higher socioeconomic class, alcohol intake and body mass index). Until adequate data are available from randomised controlled trials, the evidence-based conclusion is that HRT has no effect on breast cancer rates. HRT may reduce mortality and morbidity from other diseases and the overall risk-benefit ratio should be considered for each woman. PMID- 7476640 TI - Progress in Australia's mental health reforms. AB - The Second National Mental Health Report was released in June 1995 and documents progress in the 1993-94 financial year in implementing the five-year National Mental Health Plan. Expenditure on mental health rose by 4.1% and on community mental health services by 9.2%. State and Territory funding for non-government mental health services rose by 18% and more resources for those with disability were made available to those with psychiatric disability. The number of occupied bed-days in standalone psychiatric hospitals fell by 10%. While the report shows that Australia has moved generally in the agreed direction, considerable work remains to be done in this historically neglected area. PMID- 7476641 TI - Cytomegalovirus retinitis in patients with AIDS: current status. AB - About 20%-30% of patients with AIDS will develop cytomegalovirus (CMV) retinitis when they are severely immunodeficient (CD4 cell count below 40 cells/microL). It causes extensive retinal damage with visual deficits, and relapses are frequent. Intravenous ganciclovir and foscarnet are standard therapy, but their high toxicity and long term intravenous access mean that advances in treatment, including oral prophylactic agents, intravitreal injections and surgical implants, may provide a better quality of life for patients. Prophylactic ophthalmological screening of patients with CD4 cell counts below 100 cells/microL to detect asymptomatic retinitis is needed. PMID- 7476643 TI - Painful lessons: opioids, iatrogenic dependence and professional standards. PMID- 7476642 TI - Breast cancer: getting the diagnosis right. AB - Distinguishing between early breast cancer and benign conditions, which are much more common, can be difficult. Careful clinical examination and directed imaging, supported by fine needle aspiration cytology, provide a high level of diagnostic accuracy. PMID- 7476644 TI - Painful lessons: opioids, iatrogenic dependence and professional standards. PMID- 7476645 TI - Painful lessons: opioids, iatrogenic dependence and professional standards. PMID- 7476647 TI - Hazards of transrectal biopsy of prostate. PMID- 7476646 TI - Painful lessons: opioids, iatrogenic dependence and professional standards. PMID- 7476648 TI - Hazards of transrectal biopsy of prostate. PMID- 7476649 TI - The rate of oxygen release from single sinusoid of rat liver, determined by microspectroscopy. AB - The rate of oxygen release from single hepatic sinusoid of rat was determined. A scanning spectrophotometer, equipped with a grating and two photoncounters, was connected to a microscope with light-guides, and absorption spectra (450-650 nm) were obtained simultaneously at two microspots (10 microns diameter) on single sinusoid. The concentration ([Hb]) and oxygen saturation (SO2) of hemoglobin were calculated from the spectra. Reference transmittance was obtained at neighbouring hepatocytes. The erythrocyte velocity was measured by dual-spots cross correlation method using two photomultipliers connected to the microscope with two light-guides. The gradient in SO2 was observed along each sinusoid, due to oxygen release from flowing erythrocytes to hepatocytes. The rate of oxygen release per unit surface area was 0.24 +/- 0.14(n = 14)nmoles O2/cm2/sec, which was calculated from [Hb], difference in SO2 between the spots at up- and down stream, erythrocyte velocity, two spot's distance and sinusoidal diameter. The rates of O2 release depended on sinusoidal diameter and sinusoidal blood flow. PMID- 7476650 TI - Comparison of neural network and k-NN classification methods in medical image and voice recognitions. AB - We make a comparison of classification ability between BPN (Back Propagation Neural Network) and k-NN (k-Nearest Neighbor) classification methods. Voice data and patellar subluxation images are used. The result was that the average recognition rate of BPN was 9.2 percent higher than that of the k-NN classification method. Although k-NN classification is simple in theory, classification time was fairly long. Therefore, it seems that real time recognition is difficult. On the other hand, the BPN method is long in learning time but is very short in recognition time. Especially if the number of dimensions of the samples is large, it can be said that BPN is better than k-NN in classification ability. PMID- 7476651 TI - Pathological specimens of rat temporal bone--comparison between different fixative solution. AB - We investigated the difference in temporal bone specimens fixated by three sorts of fixative solution (10% formalin fixative, Wittmaack's fixative and Heidenhein SuSa fixative). 1. 10% Formalin's fixative solution We found many pinkish precipitates, which are stained by hematoxy-eosin, in the scala media of the cochlea. Same substance is found in the perilymphatic space and endolymphatic space in the vestibulum. The fusion of the cells is indicated in the tectorial membrane, inner sulucus cell, outer sulucus cell especially in the apical turn of the cochlea. 2. Wittmaack's fixative solution The detachment of inner and outer sulucus cells are found from basilar membrane at the basal turn of the cochlea. The tendency in convex and concave form of the Reissner's membrane is most remarkable in these three fixative solution. 3. Heidenhein-SuSa fixative solution In this solution, no precipitates, no fusion of cells are found. No detachment of the inner or outer sulucus cells is also found. This study indicates that Heidenhein-SuSa fixation is most excellent fixative method. PMID- 7476652 TI - Liver infarction produced by dual vessels occlusion due to trauma and TAE: long follow up using CT, US, and MRI. AB - The cause of liver infarction previously reported [1-4] are mainly due to occlusion of hepatic artery. Herein, we report the case of liver infarction resulted from simultaneous occlusion of hepatic artery and portal vein due to trauma and therapeutic transcatheter arterial embolization (TAE), and we followed up the infarcted lesion with computed tomography (CT), ultra sonography (US) and magnetic resonance imaging (MR imaging) until it disappeared. PMID- 7476653 TI - [Genetic technology--curse or blessing?]. PMID- 7476654 TI - [Methods for optimizing cytostatic drug therapy]. PMID- 7476655 TI - [Ovarian carcinoma--diagnosis and treatment. Consensus conference of the National Institute of Health]. PMID- 7476656 TI - [Recurrent cystitis]. PMID- 7476657 TI - [How effective is intestinal treatment of candida infection with nystatin?]. PMID- 7476658 TI - [Is abortion murder?]. PMID- 7476659 TI - [Cardiac natriuretic peptide]. PMID- 7476660 TI - [Pemphigus vulgaris. Clinical aspects, pathophysiology and therapy of a bullous autoimmune dermatosis]. PMID- 7476662 TI - [Caries prevention by fluoride tablets?]. PMID- 7476661 TI - [Optimal calcium intake. Consensus conference of the National Institute of Health]. PMID- 7476663 TI - [Changes in the approbation regulation for pharmacists?]. PMID- 7476664 TI - [Changes in the approbation regulation for pharmacists?]. PMID- 7476665 TI - Child abduction. PMID- 7476666 TI - Drink driving legislation: medicine and the law. PMID- 7476667 TI - Year and a day rule to be amended? PMID- 7476668 TI - Munchausen syndrome by proxy. PMID- 7476669 TI - Drugs for parasitic infections. PMID- 7476670 TI - Dirithromycin. PMID- 7476671 TI - Dexrazoxane for cardiac protection against doxorubicin. PMID- 7476672 TI - Melatonin. PMID- 7476673 TI - [The clinical picture and course of the nonerythematous form of ixodid tick-borne borreliosis]. AB - The territory of the Perm region is highly endemic in Ixodes (B. garinii and B. afzelii)-borne borreliosis. The clinical manifestations of erythema-less Ixodes borne borreliosis were studied in the 1991-1994 epidemiological seasons. A total of 54 patients were examined in the acute stage of infection and 4-6 months later. There were common symptoms of damage to the central nervous system in early infection and pronounced signs of damage to the central and peripheral nervous systems in the late period which was characterized by cardiovascular diseases as disturbances of automatism functions, conduction, diffuse and local muscle changes. PMID- 7476674 TI - [The presence of Borrelia in the intestines and salivary glands of spontaneously infected adult Ixodes persulcatus Schulze ticks during bloodsucking]. AB - A direct microscopic analysis of fixed smears and live preparations was used to reveal whether spirochetes are present in the gut and salivary glands of adult Ixodes persulcatus ticks spontaneously infected with Borrelia garinii and B. afzelii. Unfed ticks collected from a vegetation, partially fed ticks removed from human bodies, and ticks deliberately fed on laboratory animals were studied. In each preparation, all spirochetes were counted in 250 microscopic fields, and their concentration per 100 microscopic fields was determined. A total of 1962 ticks were individually analysed. The methods used on the study allowed a reliable identification of Borrelia in the viscera of not only unfed, but also of partially fed ticks. The infection rate in ticks that started bloodsucking was slightly lower than in the unfed ticks. This was associated with the decreased spirochete concentration in the preparations made from the partially fed ticks. Borrelia were frequently found in the salivary glands of the unfed infected I. persulcatus. During the first two to three days of bloodsucking, neither the proportion of ticks with spirochetes in the salivary glands, nor the spirochete concentrations increase. Borrelia migration from the tick gut into the salivary glands during early bloodsucking is not a prerequisite for or even important for pathogen transmission with saliva. The transmission rate appears to depend on the baseline proportion of the unfed ticks carrying spirochetes in their salivary glands.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7476675 TI - [The distribution of Trichinella nativa, T. nelsoni and T. pseudospiralis in Eurasia]. AB - A hundred and forty-six isolates of Trichinella from various regions of Eurasia have been identified by the crossbreeding method (1 male + 1 female). Three independent species of Trichinella: T. nativa, T. nelsoni, and T. pseudospiralis were found to parasite on the animals of this continent. The former two are dominant species of different territories. T. nativa occupies the whole continental zone to the north and the north-east of the isotherm minus 5-6 degrees C in January and T. nelsoni populate to the west and the south of this line. The both strains are registered in the borders of active zones of the isotherms mentioned above. T. pseudospiralis is autochthonous for the Australian region. Due to its plasticity, this species has also been found in the tropical part of Asia. Birds seem to be the carriers of this nematode to temperate latitudes. PMID- 7476676 TI - [Evidence for the transmission of the causative agent of visceral leishmaniasis by the sandfly Phlebotomus (Adlerius) turanicus Artemiev, 1974 in Turkmenistan]. AB - Four sandfly [correction of mosquito] species were identified in all foci of visceral leishmaniasis (VL) in Turkmenistan. There is evidence that three of them (Phlebotomus papatasi, P. caucasicus, P. andrejevi) cannot be carriers of VL agent in Turkmenistan. The carrier of Leishmania infantum in Turkmenistan is likely to be the zooanthropophilic sandfly [correction of mosquito] P. turanicus that prevails in the settlements and their vicinities in the desert foothill plains and in the burrows of foxes. The time of the carrier activity is May to September, and its peak size is observed in mid-July. In September, the promastigote infection of P. turanicus is as high as 47.5%. The carrier size may vary with years, the infection of children and dogs ceased with the reduction in the P. turanicus size. There are evidence for the distribution of the carrier in the populated locality, dwelling and natural biotopes. PMID- 7476677 TI - [The effect of different factors on the epidemic process in ascariasis (exemplified by Tula Province)]. AB - An epidemiological process in ascariasis in Tula Province within a 16-year period during which there were no significant socioeconomic changes is analyzed. Improved water consumption, the scope and level of waste-water disposal and sewage treatment along with intensive efforts to detect the foci of ascariasis and with the implementation of health-promoting measures in them are shown to have the most considerable influence on the reduction in the intensity of an epidemiological process in ascariasis. These findings may be useful in gaining some insight into the efficiency of health-promoting measures in the foci where sewage treatment is inadequate and water supply is poor. PMID- 7476678 TI - [The environment of the Moscow megalopolis and the problems of parasitic contamination]. AB - Studying parasitic contamination is regarded as an individual research trend for large-scale land and water areas whose natural habitat is greatly modified by man made influences. On the one hand, parasitic contamination as its particular form acts as a factor that destabilizing the environment, on the other, the man-made deformation of natural biocenoses, which is induced by socioeconomic influences promote its occurrence and manifestations. The investigations of parasitic contamination are beyond the scope of the routine parasitological goals, though they involve their whole package and should be considered as an ecological objective of environmental protection. The cenotic communications of the links of the parasitic systems with the physicochemical and biological ingredients of the habitat predetermine qualitative and quantitative differences of parasitic contamination in the large urban areas and, finally, the dissimilar course of epidemiological, epizootic, and phytototic processes in parasitic diseases in their various sectors. The major factors influencing the levels of the parasitic pollution of the Moscow megalopolis were analyzed. These included various lines of economic activities, the population's living conditions, procedures for manufacturing foodstuffs, demographic changes. In the recent years, the situation in many parasitic infections in man (trichinosis, cryptobiasis, enterobiasis, toxocariasis, etc.), animals (fascioliasis, trichinosis, etc.) and plants (viral, fungal, and nematode diseases). There is an increase in the population of mosquitos, lice, fleas, and other insects which are vectors of dangerous parasitic diseases of man and animals. PMID- 7476679 TI - [The procedure for the wide use of praziquantel in a complex of measures to control opisthorchiasis. 2. The tolerance and efficacy of Russian-made azinox in mass treatment in foci]. AB - Whether the Russian praziquantel analog azinox can be widely used in the foci of opisthorchiasis was first assessed. The outpatient treatment of 7405 patients with the agent in doses of 30, 40, 60 mg/kg body weight revealed that azinox tolerance did not depend upon the intensity and degree of clinical infection signs but it was slightly worse when a dose of 60 mg/kg was given. Children virtually showed no adverse reactions, in adults their frequency was no more than 52 +/- 1.4%. The parasitological efficiency of azinox in the used doses was 96 99% in children and 82-86% in adults. There was a clinical improvement in 65% of the treated. Thus, the national drug azinox is not inferior to the imported agent bilthricide in tolerance, parasitological and clinical efficiency. Bearing in mind the equal efficiency, but the better tolerance and less cost of the drug doses of 30 and 40 mg/kg body weight, it is advisable to use them in the foci for outpatient treatment. PMID- 7476680 TI - [The myeloperoxidase of the polymorphonuclear leukocytes as a marker of the stage of Opisthorchis invasion and as an indicator of the efficacy of chemotherapy]. AB - A total of 152 patients with Opisthorchis infection at the acute and chronic stage of the diseases, as well as 1-3 days and 1-2 months after therapy with biltricide in a dose of 60 mg/kg body weight. Intracellular myeloperoxidase was spectrophotometrically determined. Its activity was demonstrated to be significantly varying at the different opisthorchiasis stage. At the acute stage there was a 6-fold increase in the activity of the enzyme as compared with that at the chronic stage. The therapy resulted in the drop of myeloperoxidase levels, at the same time the authors interpret the heterodirectional changes in the activity of myeloperoxidase in the early post-therapeutic period as a fact of individual anthelmintic intolerance the normalization of the parameter studied 1 month after the therapy only in 42.1% of the cases as a fact residual Opisthorchis antigen persistence. PMID- 7476681 TI - [The synthesis and study of the acute toxicity and anthelmintic activity of new 2 methylthiobenzimidazoles]. AB - Several derivatives of 2-methylthiobenzimidazoles were synthesized and assayed for anthelmintic activity. Following these studies, 2-methylthio-5-(1 chloronaphthyloxy-2)-6-chlorobenzimidazole (G-1557) has been selected as a tichinellacide for detailed investigations. PMID- 7476682 TI - [The technology for manufacturing antiparasitic preparations. 4. The development of a technology for producing the anthelmintic bromoksan in finely dispersed form and the evaluation of its antifascioliasis activity]. AB - A manufacturing procedure has been developed to prepare the anthelmintic bromoxane as a finely divided dosage form. The maximum toxic dose of the drug was 1 g per kg mice body weight. Its therapeutic dose in sheep fascioliasis was 20 mg/kg. PMID- 7476684 TI - [The level of the insecticidal activity and toxicity of the new preparation Fury]. PMID- 7476683 TI - [The effect of microwaves (UHF) and magnetic fields on the insecticidal properties of biological and chemical preparations]. AB - The larvicidal activity of preparations from different groups of compounds was assayed after their exposure to microwaves and magnetic field. The radiation was found to have a varying impact on the insecticidal properties of the preparations. Thus, the efficiency of spheric larvicides decreased, which is likely to be associated with the structural changes of toxin crystals, whereas the larvicidal activity of the organophosphorus compound dursban and the pyrethroid aikon increased 2-5-fold. The higher activity of dursban irradiated with a magnetic field was demonstrated to retain for over 2 months. PMID- 7476685 TI - [The reaction of taiga ticks to an attractant. 1. The experimental contact of nymphs with attractant-acaricidal granules]. AB - Four experimental variants used the following: 1) granules only from a filler (control); 2) those from a filler and an attractant; 3) and 4) those from a filler, an attractant, and a toxicant (permethrin or lambda cigalothrin, respectively). Each experiment used 20 nymphs of the first laboratory generation. 90 and 25% were in contact with attractive and control granules, respectively. Out of the nymphs in experiments 3 and 4 granules, 94 and 100% died. The contacts of 4 sec or more duration led to death. The experimental findings suggest that the designing and application of attractive acaricidal granules in the foci of tick-borne encephalitis and Lyme's disease are promising. PMID- 7476686 TI - [Pediculosis and the problems of its control at the current stage]. PMID- 7476687 TI - [The interferon system of children with enterobiasis]. PMID- 7476688 TI - [A report of 4 cases of human Dirofilaria repens infestation in the city of Barnaul, Altai Territory]. AB - Four local cases of eye damage due to Dirofilaria repens infection which occurred in the town of Barnaul, Altai Region, are described. This information may be useful for medical and veterinary staff. PMID- 7476689 TI - [Arbovirus infections, hemorrhagic fever with renal syndrome and Astrakhan fever in the southeastern region of the Russian Federation]. PMID- 7476690 TI - [Shabsai Davidovich Moshkovskii (on the centenary of his birth)]. PMID- 7476692 TI - [Ophthalmology today worldwide and in Serbia]. PMID- 7476691 TI - [Human cercariasis caused by schistosomatid larvae from aquatic birds in the Narochanka River recreational area of Byelarus]. AB - The reasons of the dramatically aggravated cercarial situation were examined in the Naroch Lake area (Minsk Province, Byelorussia). The incidence of infection with cercariae of the family Schistosomatidae trematodes in 8 mollusk species of the genera Lymnaea, Planorbarius, Planorbis, Anisus which were nonuniformly distributed in the upper littoral were studied. The snails form mosaic congestions of various density (from solitary individuals in a 50-100 m2 area to hundreds per m2), which is associated with the nature of aquaphytocenoses and the degree of plant overgrowth. The snails infected with Trichobilharzia ocellata and Bilharziella polonica cercariae accounted for 18.8%. Schistosoma cercariae affected L. stagnalis, L. ovata, and L.p. corvus to the greatest extent. The proportion of ducks (Anas platyrhyncha) infected with the two Schistosomatidae is 30.0% in the area in question with the average infestation rate of as many as 20 trematodes of both sexes per duck. High-risk areas for human infestation (with the number of Schistosoma cercariae exceeding 10 larvae in 10 liters of water) were recorded in the upper littorals in the beach zones of all recreation institutions such as sanatoria, rest homes, camping, etc.). The fact that Schistosoma cercariae can be active in attacking man under the conditions of the Naroch Lake littoral was verified by the self-infestation of one of the authors (S.A. Be'er). Schistosome dermatitis showing its typical clinical manifestations developed after 120-130 cercariae had penetrated into the arms and legs.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7476693 TI - [The rate of polyploid fertilization of ovocytes in in vitro fertilization in patients with an increased ovarian response]. AB - Polyploid fertilization is a common cause of human embryo's early loss in in vivo (1-2%) and in in vitro fertilization (3-5%) as well. Cause of increased rate of polypoid fertilization in in vitro fertilization is multifactorial. The aim of this paper was to establish if high ovarian response to exogenous hormone stimulation of ovaries can be one of the causes of increased incidence of polyploid fertilization. The research included 125 patients from whom 568 ovocytes were obtained by ultrasound aspiratory punction of ovarian follicles. The gathered results point to the fact that exogenous ovarian stimulation and the degree of ovarian response have a significant influence on increase of polyploid fertilization's rate. Thus, in the group of patients from whom 11 or more ovocytes were obtained, the rate of polyploid fertilization amounted to 11.3%, whereas it is statistically significantly higher (p < 0.01) in regard to those in whom less important difference in the rate of polyploid fertilization considering applied schemes of ovulation's stimulation (p > 0.1) was not established nor was it established considering the age of patients (p > 0.05). Despite the fact that the rate of polyploid fertilization in in vitro conditions is higher that in the natural conception, this method of treating marital infertility opens a unique possibility to identify all irregularities considering fertilization and prevent development of such embryos on time as it happens at the very beginning of the preimplantational stage of ovum's fertilization in laboratory conditions. PMID- 7476694 TI - [The zona fasciculata of the adrenal glands 60 days after termination of exposure to chemical fertilizer]. AB - We detected fatty substances as well as enzymic activity in rats which, after a stay in workshop, where NPK fertilizer is produced, for 30 days survived another 60 days in normal conditions. The quantity of neutral fats and tryglycerides was reduced while the content of phospholipids was increased concerning the fasciculate's zone cells. After exposure to NPK fertilizer the cells nuclei got enlarged and dense. The activity of acid phosphatases increased, MAO was reduced, while the activity of succinat-dehydrogenesis was increased. The dermined quantities of fatty substances as well as the enzyme activity point to stimulated activity of fasciculate's zone cells. PMID- 7476695 TI - [Stereometric characteristics of inflammatory gingival infiltrates]. AB - The aim of this paper was to assess stereometric characteristics of gingival cell infiltrate in regard to intensity of clinical inflammation. Materials used for assessment were 70 specimen of human gingiva removed during periodontal surgery therapy. Just before the surgery in the area where specimen was to be taken, intensity of clinical gingival inflammation was determined by gingival index after Loben. Considering stereometric parameters numerical density of lymphocytes and plasma cells were established. Analyzing results it was established that by increase of intensity of clinical gingival inflammation and increase of cell infiltrate occurs, whereas an increase of numerical plasma cells' density is statistically highly important (p < 0.001), while the numerical density of lymphocytes does not changes significantly (p > 0.05). PMID- 7476696 TI - [The role of Epstein-Barr virus infections in human pathology]. AB - Epstein-Barr virus (EBV) belongs to the group of herpesvirus and may remain latent after primary infection with a possibility of periodical reactivation EBV infection is transmitted by intimate, oral contact with previously infected persons, persons sick or infected earlier which secrete virus periodically. We report on pathogenesis of primary infection as well as clinical characteristics of infective mononucleosis and other EBV associated diseases (Birkitt lymphoma, nasopharyngeal carcinoma, syndrome of chronical fatigue). PMID- 7476697 TI - [Acute normovolemic hemodilution]. AB - Performing acute normovolemic hemodilution reduces needs for transfusion of stored homologous blood, exposure of patients to homologous blood and incidence of diseases transmitted by transfusion. Apart from that it provides significant quantities of immunologically best, completely fresh blood and/or some blood components necessary for transfusion. Apart from the latest literature data, in this paper represent the first experiences considering acute normovolemic hemodilution performed in 20 patients operated for having low localized colon's neoplasm in whom amputation of rectum was performed. 780 ml of blood was exfunded from each patient with simultaneous infusion of three times larger volume of lactated Ringer's solution (In 10 patients) one volume of Haemacell (in 5 patients) and one volume of Soludex 70 (in 5 patients). Considering these patients 15600 ml autologous and 2500 ml of homologous blood was used up for transfusion. PMID- 7476698 TI - [Surgical aspects of liver resection based on 5 years' data]. AB - From 1988 to 1993 thirty liver resections were performed in thirty patients. Eight were operated for suffering from hepatocellular carcinoma, four from metastasis of colon's adenocarcinoma, five for great hemangiomas and the rest for focal liver changes. Right lobectomy was performed in six cases, as well as two trisegmentectomies and two left lobectomies while segmentectomies in the rest. Periopretive mortality occurred in two cases as two patients died because of postoperative thrombosis of vena portae and massive gastrointestinal bleeding. Recidive of malignant tumors was established in five patients after year and in other five till the second year. Benign tumors are without recidives. Later resection is a safe procedure with a considerable selection of patients, with use of latest technical conveniences, application of grafts on branches of vena portae and with routine application of intraoperative ultrasonography. The paper contains diagnostical postulates, surgical techniques and anatomic and morphologic types of resection. PMID- 7476699 TI - [Homicide and sexual psychopathology]. AB - Using data of forensic-psychiatric archive of the Institute for Neurology, Psychiatry and Mental Health in Novi Sad, 61 homicides were processed; in all cases victims and the murderers were of different sexes and were not blood kinships. By psychopathological analysis of the sample it was not possible to determine a unique profile of "sexual homicide". Considering the personality of the murderer, the degree of victimization and the importance of alcohol as a constelative factor, three categories of homicide connected with sexual life in the broadest sense were established and any degree of generalization in the practice of forensic-psychiatric theory and practice is impossible. PMID- 7476700 TI - [Immediate prosthetic reconstruction after resection of the upper jaw]. AB - In order to determine the right time when appliance should be applied to the patient after partial, subtotal or total resection of the maxilla, represent cases treated during a six year period at the Clinic of Maxillofacial Surgery of the Stomatology Faculty in Belgrade. In 11 operated patients the appliance was applied immediately after surgery and tamponade of postresection activity cavity. In 46 patients operated at other clinics it was applied more than month after surgery. Thus, 57 patients received immediate prosthetic reconstructive treatment. According to criteria of adaptability considering new appliances major differences could be observed among different groups of patients. 57 patients recovery was normal, while in 4 adaptability was slower. In 12 referral cases not a single case of normal adaptability was registered. 4 patients demonstrated slower adaptability, while in 8 patients severe adaptability problems were registered. PMID- 7476701 TI - [Comparative value of skin tests and IgE antibodies in the serum in patients with chronic vasomotor rhinitis]. AB - In order to assess values of skin testing and determination of pulp, as a credible indicator considering sensibilisation of an organism, 61 patients suffering from chronic vasomotor rhinitis were tested. By application of "prick" test, skin test results were positive in 30 patients and negative in 31. Presence of specific IgE antibodies in the serum was assessed in all patients by "ELISA" test. In 51 patients (83.6%) both kinds of findings corresponded. In 26 patients both skin parameters were negative while in 10 patients (16.40%) findings did not correspond. It was established that for Dermatophagodies pteronyssinus specific IgE antibodies are determined in serum when pap is 5 mm wide. Grasp pollen always caused strong local reactions, regularly more than 10 mm. Considerably high levels of IgE antibodies was also established while other pollens did not show such correspondence. PMID- 7476702 TI - [Facial hemiatrophy--Romberg's disease]. AB - Author present a care of acquired facial hemiatrophy--Romberg's disease, a sporadic disease of unclear etiology characterized by shrinking and deformation of one side of face, including superficial facial tissues, subcutaneous tissues, fatty tissue, musculature, cartilage and bones. The principle etiopahogenetic hypothesis suggests that Romberg's disease is the consequence of central and peripheral alterations in the sympathetic nervous system. The disease was first described by Romberg in 1846, and in 1963 Roger gave a detailed description and analysis of 1035 cases. In this study authors presented the latest information form literature (magnetic resonance image, biochemical investigations, experimental model of a sympathectomy at animals). A case of a patient treated with great number of available methods: computerized tomography, X-ray, Doppler spectral analyze, termovision, electromyography, electroencephalography, evoked visual potentials, brainstem's evoked potential is shown. PMID- 7476703 TI - [The effect of hallucinogenic drugs on the development of psychoses]. AB - Psychotic behavior and suicide which occurred after it induced us to study a connections among LSD use, psychotic behavior and suicidal risk ones again. Hereby we have a case of a young man who showed a psychotic behavior a week after taking LSD which ended tragically with suicide. What was the cause of this action, was it a late consequence of drug intake, flush-back or the first episode of a dissociative process which possibly started by change of behavior and sphere of interest? It is hard to answer this question by psychological autopsy. The dilemma remains constant, but we still think that this case report is of importance because we must not neglect such a clinical picture after taking LSD. It is particularly important now as drug addition is abruptly increasing in our society. PMID- 7476704 TI - [The effect of marriage on mental health. Dr. Djordje Natosevic: psychological aspects of marriage]. AB - Dr. Dorde Natosevic (1821-1887), our famous physician, health educator and reformer of the educational system, published an article "Marriage in regard to psyche" in the 111, book of the Chronical of Matica Srpska in 1866. In his article he consider the influence of both married and unmarried life on mental health of men and women. He gives advice: when, how and with whom one should and with whom one should not get married. His special interest were unmarried women and he gave numerous advice about making there lives more beautiful and interesting. Some of these references are actual even these days. PMID- 7476705 TI - [Lipid status in extreme obesity]. AB - 50 extremely obese women < 35 years of age (their height 164 cm, weight 107 kg, BMI 40) were analyzed in this paper. Following lipidic and lipoprotein parameters were determined: total triglycerides, total HDL-, LDL-cholesterol, electrophoresis of serum's lipoproteins, and in the third of patients the level of apoproteins. AI, AII and B. In all patients the level of glycemia on an empty stomach was determined. Gathered results were compared with those of the control group which consisted of 32 metabolically healthy women of corresponding age. On the basis of gathered results it can be concluded that in extremely obese women the level of numerous atherogenic lipidic fractions such as total triglycerides, total cholesterol, LDL-cholesterol and apoprotein B was statistically significantly increased, while the level of protective HDL-cholesterol was statistically significantly decreased. Results of this investigation point to the common connection between obesity and disorder of metabolism considering lipids and lipoproteins which result in increased incidence of cardiovascular diseases in obese persons. PMID- 7476706 TI - Influence of blood vessels on the measurement of hemoglobin oxygenation as determined by time-resolved reflectance spectroscopy. AB - We report the development of a heterogeneous resin-tube model to study the influence of blood vessels on the apparent absorption of the system, mu a(sys), using a time-resolved technique. The experimental results show that mu a(sys) depends on the absorption inside the tubes, mu a(tube), tube diameters, and tube to-sample volume ratios. A mathematical expression relating mu a(sys) and mu a(tube) is derived based on the experimental results and is verified by time resolved Monte Carlo simulations for heterogeneous models. This analytical formula predicts that the apparent absorption coefficient measured on a biological organ is a volume-weighted sum of the absorption coefficients of different absorbing components. We present some apparent absorption coefficients measured in vivo in animals and humans and discuss improved algorithms that calculate the hemoglobin saturation by including background-tissue absorption and blood vessel distribution. PMID- 7476707 TI - Determination of 3D imaging geometry and object configurations from two biplane views: an enhancement of the Metz-Fencil technique. AB - We present a new technique based on the method developed by Metz and Fencil for estimation of the 3D imaging geometry and 3D object configurations from biplane angiographic acquisitions. The new method employs the 3D configuration of points calculated by the Metz-Fencil technique as an initial estimate. A 3D Procrustes algorithm is employed to translate, rotate, and scale the configuration until it aligns optimally with the set of lines that connects a focal spot with the corresponding set of image points. This alignment procedure is applied independently for each view. The rotation and translation that relate the two aligned data sets are then determined by an additional 3D Procrustes calculation. These steps are applied iteratively. Evaluations were based on Monte Carlo simulation and phantom studies. With this new technique, the mean absolute errors in magnification, in the relative position of the points, and in the angles defining the rotation and translation matrices were approximately 3.0%, 1.5 mm, and 5 degrees and 3 degrees, respectively, for rms input errors in the image data up to 2.0 pixels (0.7 mm). Errors in the results can be as small as 0.5%, 0.16 mm, 0.6 degrees, and 0.3 degrees, respectively, if input image-data error is 0.035 mm. The improvement of the Metz-Fencil technique described here may provide a basis for precise estimation of the biplane imaging geometry and the 3D positions of vessel bifurcation points. PMID- 7476708 TI - The effect of fat on the coherent-to-Compton scattering ratio in the calcaneus: a computational analysis. AB - The coherent-to-Compton scattering ratio (CCSR) is a technique that has been proposed for measuring trabecular bone mineral density (TBMD). This paper investigates the effect of fat on the CCSR and its correlation to the error in TBMD measurements. It is a computational study to determine the relationship between the magnitude of fat error and the momentum-transfer variable chi, which represents the incident photon energy and the scattering angle. Variation in fat content contributes significantly to the error in CCSR measurements. When employing a typical 241Am source (E gamma = 59.45 keV), the resulting error decreases with increasing momentum-transfer variable or angle. For example, the error ranges from +14 mg/cc at an angle of 45 degrees (chi = 18.3) to +3 mg/cc at an angle of 135 degrees (chi = 44.3) for an osteoporotic trabecular region (100 mg/cc mineral) of a calcaneus that contains 6% less fat than a calibration standard. The error is about 0.3-1.2 mg/cc less for regions containing 2-3X more bone mineral and is reduced and opposite in sign for regions containing about 7% more fat than the calibration standards (e.g., -9 mg/cc at 45 degrees and -1.5 mg/cc at 135 degrees). Others have shown that the intrinsic sensitivity of the CCSR method for measuring TBMD at a given photon energy generally increases with increasing detector angle. Thus large angles are advantageous both for reduced sensitivity to fat variation and increased sensitivity to bone mineral variation. The primary disadvantage is reduced count rates that degrade precision unless long counting lines are employed.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7476709 TI - The spectrum and angular distribution of x rays scattered from a water phantom. AB - To calculate the response of an image receptor to the x rays emerging from a scattering medium, it is necessary to know the x-ray spectrum and intensity as a function of the angle of incidence on the receptor. To permit this calculation for any x-ray spectrum incident on a medium, these functions must be known for monoenergetic x rays. For monoenergetic x rays in the range 20-70 keV we have measured with a high-purity germanium detector the spectrum and intensity of x rays emitted from a water phantom at angles of 0 degree-50 degrees to the direction of the primary beam. The spectrum and intensity of emitted x rays have also been calculated by the Monte Carlo method. At small exit angles, most of the x rays have energies close to the incident energy. As the exit angle increases, the fraction of multiply scattered x rays increases. At very large exit angles, the dominant feature of the spectrum is the peak due to these multiply scattered x rays. For small scattering angles the Monte Carlo calculations are in good agreement with the measurements over the range of energies. For large scattering angeles the scattered photon fluence predicted by Monte Carlo modeling is consistently lower than the measurement in the region just below the full energy peak. The cause of the discrepancies is not fully understood, but cannot be accounted for by Compton broadening alone. An alternate approach to model incoherent scattering is proposed. PMID- 7476710 TI - Tree structured wavelet transform segmentation of microcalcifications in digital mammography. AB - A novel multistage algorithm is proposed for the automatic segmentation of microcalcification clusters (MCCs) in digital mammography. First, a previously reported tree structured nonlinear filter is proposed for suppressing image noise, while preserving image details, to potentially reduce the false positive (FP) detection rate for MCCs. Second, a tree structured wavelet transform (TSWT) is applied to the images for microcalcification segmentation. The TSWT employs quadrature mirror filters as basic subunits for both multiresolution decomposition and reconstruction processes, where selective reconstruction of subimages is used to segment MCCs. Third, automatic linear scaling is then used to display the image of the segmented MCCs on a computer monitor for interpretation. The proposed algorithms were applied to an image database of 100 single view mammograms at a resolution of 105 microns and 12 bits deep (4096 gray levels). The database contained 50 cases of biopsy proven malignant MCCs, 8 benign cases, and 42 normal cases. The measured sensitivity (true positive detection rate) was 94% with a low FP detection rate of 1.6 MCCs/image. The image details of the segmented MCCs were reasonably well preserved, for microcalcification of less than 500 microns, with good delineation of the extent of the microcalcification clusters for each case based on visual criteria. PMID- 7476711 TI - Bayesian image estimation of digital chest radiography: interdependence of noise, resolution, and scatter fraction. AB - Previously, it has been shown that Bayesian image estimation (BIE) can reduce the effects of scattered radiation and improve contrast-to-noise ratios (CNR) in digital radiographs of anthropomorphic chest phantoms by improving contrast while constraining noise. Here, the use of BIE as a noise reduction technique is reported. An anthropomorphic phantom was imaged with a previously calibrated photostimulable phosphor system using standard bedside chest radiography protocols. The Bayesian technique was then used to process this image. BIE incorporates a radial exponential convolution scatter model with two adjustable parameters. In previous reports, these parameters were optimized to reduce the residual fraction of scattered radiation in the processed image. Here, the parameters were adjusted to evaluate the potential of BIE to reduce image noise. While the full width at half maximum of the scatter model was held constant, the magnitude was varied. Evaluation was based on residual scatter fractions and CNR. The magnitude of the kernel in the scatter model was varied from 0.0 to 2.5 in steps of 0.5. Previously, it was found that an "ideal" scatter kernel magnitude of 2.33 provided a minimum residual scatter fraction. This magnitude corresponds to the average scatter-to-primary ratio in the chest radiograph. As the magnitude was increased, the residual scatter fraction decreased and the CNR increased in both the lungs and the mediastinum. However, as the magnitude was decreased, the percent noise also decreased; therefore, a lower magnitude kernel reduces noise. By varying the magnitude of the kernel used, differing amounts of noise reduction and contrast enhancement can be obtained.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7476712 TI - Energy imparted in computed tomography. AB - Monte Carlo techniques were used to study a generalized CT dose index D(r) as a function of the radius r of a cylindrical dosimetry phantom. The relationship between D(r) and the energy deposited in the phantom was investigated. For a specific x-ray spectrum, the energy imparted to head or body dosimetry phantoms can be obtained from measured D(r) values. This approach to CT dosimetry permits the energy imparted to phantoms (or patients) to be determined as CT technique parameters, or type of scanner, are changed. PMID- 7476713 TI - Experimental validation of an elementary formula for estimating spatial resolution for optical transillumination imaging. PMID- 7476714 TI - A fast and stable maximum a posteriori conjugate gradient reconstruction algorithm. AB - We have derived a maximum a posteriori (MAP) approach for iterative reconstruction based on a weighted least-squares conjugate gradient (WLS-CG) algorithm. The WLS-CG algorithm has been shown to have initial convergence rates up to 10x faster than the maximum-likelihood expectation maximization (ML-EM) algorithm, but WLS-CG suffers from rapidly increasing image noise at higher iteration numbers. In our MAP-CG algorithm, the increasing noise is controlled by a Gibbs smoothing prior, resulting in stable, convergent solutions. Our formulation assumes a Gaussian noise model for the likelihood function. When a linear transformation of the pixel space is performed (the "relaxation" acceleration method), the MAP-CG algorithm obtains a low-noise, stable solution (one that does not change with further iterations) in 10-30 iterations, compared to 100-200 iterations for MAP-EM. Each iteration of MAP-CG requires approximately the same amount of processing time as one iteration of ML-EM or MAP-EM. We show that the use of an initial image estimate obtained from a single iteration of the Chang method helps the algorithm to converge faster when acceleration is not used, but does not help when acceleration is applied. While both the WLS-CG and MAP-CG methods suffer from the potential for obtaining negative pixel values in the iterated image estimates, the use of the Gibbs prior substantially reduces the number of pixels with negative values and restricts them to regions of little or no activity. We use SPECT data from simulated hot-sphere phantoms and from patient studies to demonstrate the advantages of the MAP-CG algorithm. We conclude that the MAP-CG algorithm requires 10%-25% of the processing time of EM techniques, and provides images of comparable or superior quality. PMID- 7476715 TI - Magnetically enhanced protection of bone marrow from beta particles emitted by bone-seeking radionuclides: theory of application. AB - Utilization of radiopharmaceuticals that directly target radioactivity to tumors for treatment has a great deal of promise. Ideally, lethal doses of radiation could be delivered precisely to areas of disease, while, for the most part, sparing normal tissues. This potential, however, has not yet been fully realized. Current limitations of this approach are low tumor uptake of radiopharmaceuticals and dose-limiting radiotoxicity. In an effort to offset low uptake, radionuclides that emit high average-energy electrons have been proposed. Unfortunately, use of these radionuclides increases myelosuppression on a per decay basis. In order to allow for the utilization of high doses of this class of high-energy beta emitters, we propose the application of a strong static homogeneous magnetic field to constrain the beta particles. Monte Carlo computer simulations indicate that application of a 10 T magnetic field can decrease the total radiation dose from bone-avid tracers to marrow located in shafts of human long bones by 14%. More significantly, however, the penetration depth of high-energy electrons from the bone surface into the marrow can be reduced by up to 74.6%. Preservation of marrow in areas distal to the bone has previously been shown to facilitate relatively rapid recovery from pancytopenia produced by radiation damage to trabecular marrow (without marrow transplantation). Magnetically enhanced protection of bone marrow, therefore, may allow administered doses of high-energy beta-emitting radionuclides to be increased. By raising the limits on injected quantities of such highly ionizing radionuclides, amounts of the radiation dose absorbed by both soft and calcified tissue tumors will be increased, compared to conventional treatments.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7476716 TI - Proton activation analysis of stable isotopes for a molybdenum biokinetics study in humans. AB - Molybdenum is a trace element essential to life. Nevertheless, little information is available on its metabolism in humans. A methodology based on stable isotope administration that combines compartmental analysis, simultaneous use of two tracers, and proton nuclear activation (PNA) is presented. A four-compartment metabolic model was adopted. The compartments are stomach, small intestine, transfer compartment, and unquantified tissue pool. The employment of two different stable isotopes of the element under investigation as tracers was made possible by PNA. Optimization of the technique for molybdenum determination in plasma led to the choice of 95Mo and 96Mo as tracers. Their concentrations in plasma can be determined measuring the disintegration gamma lines of the corresponding technetium radioisotopes produced via (p,n) reaction. In the adopted experimental conditions, a minimum detectable concentration of 2 ng isotope/ml plasma was attained. A kinetics study was performed on two healthy volunteers. To both subjects one tracer was orally administered, and the other intravenously injected. Venous blood samples were withdrawn at different postinjection times and the concentrations for both isotopes determined. The model parameters describing molybdenum kinetics were obtained for the two individuals. Total absorbed fraction was found to be 0.84 +/- 0.03 and 0.86 +/- 0.07, respectively. PMID- 7476718 TI - The calibration and use of plane-parallel ionization chambers for dosimetry of electron beams. AB - The AAPM TG 39 protocol has proposed three different methods of calibrating plane parallel ionization chambers, i.e., in-phantom irradiation with a high-energy electron beam and in-phantom and in-air 60Co irradiation. To verify the consistency of the three methods, we have measured Ngaspp values using each of these techniques for the five most commonly used plane-parallel chambers considered by the protocol. Our results demonstrate that the measured Ngaspp values for the three different methods for any of the chambers agree to within +/ 0.6%. Once Ngaspp was measured, the determination of absorbed dose for electron beams with different energies for an AECL Therac 20 and Philips SL25 was carried out according to the AAPM TG 39 protocol. The results show that the determination of the absorbed dose outputs for any of the five chambers agree to within +/- 0.7% for electron-beam energies of 4-20 MeV if all five chambers had Ngaspp values determined by the electron-beam method. The uncertainties are well within the expected error for these approaches. PMID- 7476717 TI - A method to evaluate tracer kinetics in small laboratory animals using a series of thermoluminescent dosimeters. AB - Miniature detector probes have previously been used in large animal models to investigate myocardial 201Tl clearance kinetics. The results of these studies helped develop clinical imaging protocols that greatly improved the accuracy of thallium scintigraphy. However, miniature detector probes are too large to be used in small animals. Thus, if a method could be developed to measure regional time activity curves in small animals, it would provide a cost-effective alternative to both experiments in large animals and/or multiple experiments at varying time points that can produce results only by postmortem analysis of several animals. Accordingly, we developed a method to measure a regional time activity curve of a tracer in rabbits by using a series of thin thermoluminescent dosimeters [CaF2 (dopant) TLDs, 1 mm thick] placed on the surface of the myocardium. Background contributions associated with high blood pool activity are modeled and then subtracted from the initial TLD response. To validate and illustrate this method, thallium kinetics were determined for nonischemic rabbit myocardium (n = 6). Myocardial thallium concentration decreased monoexponentially with a mean half-time equal to 396 +/- 141 min. Arterial blood activity decreased triexponentially with a final half-time of 243 +/- 73 min. No significant difference was found when the myocardial half-time was compared to the final arterial half-time. These findings are consistent with previous work using a cadmium telluride probe in a canine model. Therefore, TLD analysis can provide a cost-effective, reliable, and reproducible method to measure regional myocardial clearance kinetics. PMID- 7476719 TI - A convolution-adapted ratio-TAR algorithm for 3D photon beam treatment planning. AB - A convolution-adapted ratio of tissue-air ratios (CARTAR) method of dose calculation has been developed at the Mallinckrodt Institute of Radiology. This photon pencil-beam algorithm has been developed and implemented specifically for three-dimensional treatment planning. In a standard ratio of tissue-air ratios (RTAR) algorithm, doses to points in irregular field geometries are not adequately modeled. This is inconsistent with the advent of conformal therapy, the goal of which is to conform the dose distribution to the target volume while sparing neighboring sensitive normal critical structures. This motivated us to develop an algorithm that can model the beam penumbra near irregular field edges, while retaining much of the speed for the original RTAR algorithm. The dose calculation algorithm uses two-dimensional (2D) convolutions, computed by 2D fast Fourier transform, of pencil-beam kernels with a beam transmission array to calculate 2D off-axis profiles at a series of depths. These profiles are used to replace the product of the transmission function and measured square-field boundary factors used in the standard RTAR calculation. The 2D pencil-beam kernels were derived from measured data for each modality using commonly available dosimetry equipment. The CARTAR algorithm is capable of modeling the penumbra near block edges as well as the loss of primary and scattered beam in partially blocked regions. This paper describes the dose calculation algorithm, implementation, and verification. PMID- 7476720 TI - Parametrization of head-scatter factors for rectangular photon fields using an equivalent square formalism. AB - Head-scatter factors of symmetric square and rectangular fields (field center on the central beam axis) defined by the upper (Y) and lower (X) jaws for 6 and 15 MV photon beams from 2300CD and 600C accelerators (Varian Associates, Inc., Palo Alto, CA) were measured, as well as those for fields shaped by the Y jaws and the multileaf collimator (MLC) of the 2300CD. For rectangular fields, the head scatter factor for the field (x = a and y = b) was different from that for the field (y = a and x = b). This difference was 2% -3% for fields defined by conventional collimators when [formula, see text] was large, and became 4%-5% when the MLC and YT jaws were used to shape the fields with the X jaws retracted. In order to calculate values for head-scatter factors of rectangular fields accurately using an equivalent square formalism, the side of the equivalent square should be obtained with different weights for lower and upper jaws, as proposed by Vadash and Bjarngard [Med. Phys. 20, 733-734 (1993)]. Our measurements demonstrate that the relative weight (G) of upper and lower jaws is strongly dependent on their distances from the x-ray source, while the beam energy has little effect on the value of G. We further show that G can be calculated simply from these distances. An analytical representation for head scatter factors of square and rectangular fields is also developed in this paper. The quality of this representation was judged by the root-mean-square (rms) deviation from measured head-scatter factors, which ranged from 0.11%-0.27%. PMID- 7476721 TI - An evaluation of the recommendations of the TG-25 protocol for determination of depth dose curves for electron beams using ionization chambers. AB - AAPM Radiation Therapy Committee Task Group 25 has recently outlined a protocol for the determination of relative dose curves for electron beams [Med. Phys. 18, 73-109 (1991)]. We have performed an evaluation of this protocol by comparing the central axis depth dose curves determined from measurements using two different ionization chambers and three different phantom materials. Measurements were made with a Farmer-type PTW and Capintec ionization chamber in solid water, PMMA, and clear polystyrene phantoms irradiated by 6- and 15-MeV electron beams. Central axis depth dose curves were generated from the measured depth-ionization data using the new protocol. For both the chambers and energies investigated in this study, excellent agreement was observed among all the depth doses in water obtained from measurements in all of the three phantoms studied. PMID- 7476722 TI - Scattered photons from wedges in high-energy x-ray beams. AB - The presence of a wedge increases the fraction of "head-scattered" photons in a high-energy x-ray beam. We have compared internal and external wedges for x-ray beam energies between 6 and 25 MV by determining their SPRw, i.e., the ratio of the dose contribution from photons scattered by the wedge and photons either coming directly from the target or scattered by other structures, including the flattening filter. Marked differences were observed. First, SPRw for the thickest external wedge (60 degrees) was 1.8% at a field size of 10 x 10 cm2 and mildly dependent on the photon energy, while SPRw for internal wedges for this field size varied between 4.4% and 5.4% depending mostly on the location and size of the wedge and marginally on the photon energy. Second, the variation of SPRw with the collimator setting c x c was different for the internal and external wedges. SPRw for the internal wedge approached a limiting value at large c and could be fitted with an error function, while SPRw for the external wedge increased quadratically with c. As a result, the difference in SPRw (c) for internal and external wedges is reduced for large fields. PMID- 7476723 TI - A QA phantom for dynamic stereotactic radiosurgery: quantitative measurements. AB - A spherical acrylic phantom was designed for quality assurance measurements of dynamic radiosurgery. The phantom consists of two mating hemispheres mounted on a base plate. The interhemispheric plane may be oriented at any angle to the base, the angle being identified by visible marks on the base plate of the phantom. The phantom has a set of replaceable, radiologically identifiable markers, suitable for Computed Tomography (CT), Magnetic Resonance (MR), and Digital Subtraction Angiography (DSA) imaging. The frame coordinates of each marker are calculated from its known positions with respect to the center of the sphere. The measured errors of these positions using CT and MR images, were within the voxel size of the displayed image, while for DSA images the error was greater than 2.5 mm at the periphery of the image. The calculated depths from the planning software, for various beam intersection points to the isocenter, agreed within 0.6 mm with the known depths. A variation of 3.6 +/- 2.6 mm in the calculated depths was observed between using MR and CT image data. This difference results in a 1% variation in Tissue Maximum ratio (TMR) calculations. Comparisons of measured and known volumes resulted in differences of 8%-10%. PMID- 7476724 TI - Source localization for brachytherapy implants loaded with an afterloader. PMID- 7476725 TI - Comments on "Dosimetry of interstitial brachytherapy sources: recommendations of the AAPM Radiation Therapy Committee Task Group No. 43" [Med. Phys. 22, 209-234 (1995)]. PMID- 7476726 TI - [Regional chemotherapy in advanced pancreatic carcinoma]. AB - BACKGROUND: Even after resection median survival times in advanced pancreatic carcinoma are seldom more than one year. The aim of our study was to evaluate the effectivity of regional chemotherapy in patients suffering from unresectable pancreatic cancer. PATIENTS AND METHODS: From 2/92 until 11/94 32 patients with unresectable pancreatic cancer (17 patients stage III, 15 patients stage IV) were treated by celiac trunc infusion. One cycle consisted of mitoxantrone (d 1), 5 FU, folinic acid (d 2 to 4) and cisplatinum (d 5). RESULTS: Besides a few severe complications, therapy was well tolerated in 101 cycles. In unresected patients with stage III tumors the median survival is 12 months compared to 4.8 months in patients with biliodigestive bypasses (p < 0.006) and 4 months in regionally infused patients stage IV compared to 2.7 months in untreated patients (n.s.). CONCLUSION: Regional chemotherapy in advanced pancreatic carcinoma is well tolerated and increases median survival in patients with stage III tumors. PMID- 7476728 TI - [Chronic pancreatitis]. PMID- 7476727 TI - [Tuberous sclerosis: typical complications of a hereditary disease and therapeutic options]. AB - BACKGROUND: Tuberous sclerosis is an autosomal-dominant hereditary disease (incidence and prevalence 1:10,000) which is characterized by hamartomas in various organs. PATIENTS AND METHODS: We demonstrate three patients with different and partly atypical manifestations: All had facial angiofibroma. No patient was mentally retarded and epilepsy was observed only intermittently. In all cases periventricular calcifications were noted and renal angiomyolipomas were found. Two patients requested dialysis after nephrectomy. One patient suffered from severe complications of the rare pulmonary lymphangioleiomyomatosis. RESULTS: Therapy of this multiorgan disease is only symptomatic. Operations should be considered as second line treatment. In one case, successful renal transplantation is described. CONCLUSIONS: Genetic counselling is warranted in this disease with high penetrance, particularly the broad spectrum of manifestations should be taken into account. PMID- 7476729 TI - [Recommendations for structured patient training in obstructive respiratory tract diseases. Patient Education Study group of the German Society of Pneumology and the German Respiratory Tract League of the German Society of Pneumology]. PMID- 7476730 TI - [Intravenous immunoglobulins. Basic principles, selection criteria and indications for preventive and therapeutic administration]. PMID- 7476731 TI - [Chronic myeloid leukemia--latest aspects and practical progress in recent years]. PMID- 7476732 TI - [Treatment of portal hypertension]. PMID- 7476733 TI - [Recommendations of the German Society of Clinical Pharmacology and Therapy for implementing and evaluating pharmaco-economic studies]. PMID- 7476734 TI - [Giant cell arteritis--a rare cause of severe arterial occlusive disease of the leg arteries]. PMID- 7476735 TI - [Talking medicine contra alternative medicine?]. PMID- 7476736 TI - Sedation with benzodiazepines. PMID- 7476737 TI - Transesophageal echocardiography in the intraoperative evaluation of hemodynamic status. PMID- 7476739 TI - Metabolic management of diabetes during labor and delivery. PMID- 7476738 TI - Hemodynamic and oxygenation variables during radical cystectomy. Does the anesthetic technique really matter? AB - The effect of anesthetics on hemodynamic variables (HV) has been clarified, but ambiguity existed concerning their effect on oxygenation variables (OV). Radical cystectomy provided a clinical setting for studying the effect of anesthetics on perioperative HV and OV. Patients subjected to radical cystectomy (n = 33) were assigned through balanced randomization to receive one of four anesthetic modalities, namely; group I: inhalation anesthesia using N2O:O2, halothane, d tubocurarine (n = 11); group II: inhalation anesthesia using N2O:O2, halothane, d tubocurarine, and supplemented with epidural analgesia (EA) (n = 11); group III: total intravenous anesthesia (TIVA) using ketamine 10-30 ug.kg-1.min-1, propofol 2 mg.kg-1.h-1, d-tubocurarine, and supplemented with continuous EA (n = 6): and group IV:TIVA using ketamine 20-50 ug.kg-1.min-1, midazolam in increments of 1.5 to 5 mg, and supplemented with intermittent EA (n = 5). Monitoring entailed continuous ECG, pulse oximerty, invasive arterial pressure, and pulmonary artery catheter for HV (HR, MAP, PAP, PAOP, CO, SVR, and PVR) and OV. (PaO2, SaO2, PvO2, SvO2, a-vDO2, O2ext, Qs/Qt, DO2, and VO2). The heart rate was lower in TIVA while other HV did not show striking differences, Group I showed higher arterial oxygen tension than group II and IV. Mixed venous oxygen tension and saturation were higher in group I over group IV. Other OV did not show remarkable differences. In conclusion, HV and OV in 4 anesthetic modalities did not elicit striking differences. PMID- 7476740 TI - Pipecuronium bromide: a new steroid muscle relaxant--a clinical study. PMID- 7476741 TI - Effect of preoperative oral fluids on gastric volume and pH in postpartum patients. AB - The effect of preoperative oral fluids to postpartum patients on the gastric volume and pH was compared with normally fasting postpartum and non-pregnant patients undergoing sterilisation. Forty consecutive postpartum patients and twenty non-pregnant patients scheduled for sterilisation procedure were selected for the study. All patients were fasted overnight. Twenty postpartum patients were randomly allocated to receive 150 ml of plain water two and half hours before surgery (Group 1), while the remaining postpartum patients (Group 2) and non-pregnant patients (Group 3) continued their overnight fast as usual. After induction of anesthesia, gastric volume and pH was measured in all the patients. The mean gastric volume was found to be 21.9 +/- 8.49 ml in Group 1, 22.55 +/- 8.30 ml in Group 2 and 22.65 +/- 8.17 ml in Group 3. The mean pH was found to be 2.21 +/- 0.98 in Group 1, 2.18 +/- 0.88 in Group 2 and 2.12 +/- 1.02 in Group 3. The difference in the volume and pH in all the three groups was statistically insignificant. The difference in the incidence of patients with combined high risk factors of acid aspiration syndrome (gastric fluid volume > 25 ml and pH < 2.5) among the three groups was also insignificant. The authors conclude that ingestion of 150 ml of plain water approximately two and half hours before scheduled time of surgery in overnight fasted postpartum patients does not increase the risk of aspiration syndrome. PMID- 7476742 TI - Ranitidine versus cimetidine prior to emergency obstetric anesthesia. AB - Twenty parturients in labour received emergency obstetric anesthesia were randomly divided into two equal groups. Group "R" received 150 mg oral ranitidine tablet on admission, followed by 50 mg infusion in 250 ml dextrose 5% over 30 minutes prior to anesthetic induction. Group "Ce" received 400 mg cimetidine oral tablet and 100 mg infusion in 250 ml dextrose 5% over 30 minutes. Ten parturients were considered as control. Ranitidine significantly reduced the maternal gastric volume with marked alkalinization of gastric pH. No significant changes were detected in the height, frequency or amplitude of uterine contraction or neonatal assessment. PMID- 7476743 TI - The anesthetic and intensive care management of pulmonary hemosiderosis in a child--case report. AB - Pulmonary hemosiderosis represents challenges during anesthesia and intensive care management. This is a report of a case of idiopathic pulmonary hemosiderosis in a child which was successfully managed during anesthesia and in the intensive care unit with total remission of the disease. The management of this and similar cases are discussed. PMID- 7476744 TI - Respiratory complications in children emerging from halothane anesthesia--awake vs deep extubation. PMID- 7476746 TI - [Intestinal intussusception in children]. AB - Enteric intussusception is one f the most frequent causes of acute abdomen in early childhood, with an incidence of 1.3-2/1000 children born and higher frequency from the third to the twelfth month. Primary intussusception is related to predisponsing factors such as peristalsis disorders or Peyer's patch hypertrophy induced by viral infection. Secondary intussusception is due to organic injury in the intestinal wall. The most involved sites are the terminal ileum and the ileocecum, the most frequent type is ileocolic intussusception. Many clinical forms exist, including acute enteric intussusception with its pathognomonic triad intermittent abdominal pain, emesis and rectal bleeding and the atypical form with a neurological presentation, where sopor, myosis and muscular atonicity are dominating, Intussusception can also present in a subacute or chronic form with a slow and apsecific onset. In a retrospective investigation we examined 30 cases of intussusception in children hospitalized at the Pediatric Clinic of Pisa from the 1960s up to today. Our patients (16 males and 14 females) were aged between one month and two years. Clinical presentations resulted in; typical forms (60%), atypical forms (16%), subacute forms 13%) and recurrent forms (10%). Clinical suspicion was confirmed either by the presence of blood observed during rectal exploration, which is a pathognomonic sign, or by the opaque enema which led to recovery by means of hydrostatic reduction in 40% of the cases. The remaining patients (60%) underwent surgery.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7476745 TI - [Management of congenital nasolacrimal duct obstruction. Timing of probing]. AB - In the management of congenital nasolacrimal duct obstruction (CNDO), there is not a well-established agreement about the proper time for probing. Some authors perform it prior to the first year of life, since after this age the relapse frequency would be very high; others prefer conservative management, since they deny this high frequency. In order to evaluate which one of these two opinions has to be considered more appropriate, the present study retrospectively reviewed the frequency of relapses at the 6-month-mark after surgery in 77 patients (40 males, 37 females) probed for CNDO at the Children's Hospital of Trieste in the 1990-1992 period. In our case-record relapses were found in 7.8% of all patients (8.8% of eyes), while the corresponding percentages were 10.8% of patients (14.3% of eyes) in those probed in the first year of life, 5% of patients (3.8% of eyes) in those probed after this time. These results show no worsening for late probing outcome, with respect to early probing. As a consequence of these findings, it can be recommended that for CNDO it is proper to start with a conservative management, represented by lacrimal sac massage and antibiotic eye drops instillation, if mucopurulent discharge is present, for a 4-6 weeks period. If symptoms do not disappear completely after this period, whether probing is the treatment of choice, possibly after the 6th month of life, or, when parents prefer this option, to resume the medical treatment, since also late probing guarantees a good outcome. The choice for probing is mandatory after second year of life. PMID- 7476747 TI - [Object permanence in children with neurological and psychomotor disorders]. AB - The aim of this research was to investigate whether the development of object permanence is an available sign of the cognitive development in infants with psychomotor handicaps. Subjects consisted of 5 males, ranging in age from 12 to 33 months, with handicaps as a result of perinatal brain injury. Four were preterm infants. All of the children received psychomotor treatment. Their stage of object permanence was assessed using traditional (B-L) and non traditional test (U-H). (The children's performances on the U-H scales have an "Intrinsic validity"). Four children achieved the first stages of the development of object permanence. The acquisition of object permanence was delayed in comparison with the age-appropriated time, but it may be considered adequate in comparison with the "developmental age" (B-L). One children with severe mental and motor disorders solved no task, the findings suggest that the children with psychomotor handicaps may gain the concept of object permanence and that psychomotor treatment may assist them in the development of the concept. This performance is not age-dependent. So the acquisition of the concept of object permanence may be considered an accurate and sensitive tool for the follow-up of the sensorimotor development in the handicapped children. PMID- 7476748 TI - [On the diagnosis of so-called "constitutional" bone diseases]. AB - The authors review the diagnosis process of so-called "constitutional" bone diseases. They criticize the nomenclature and international classification of them and suggest a diagnostic classification which is virtually independent of structural and metabolic criteria: based on the site and extent of pathological manifestations. The paper also examines the term "dysmorphia", given that it is the shape of the bone segment which initially attracts and guides the clinical diagnosis of embryonal osteochondropathies and suggest it. PMID- 7476749 TI - [Early myoclonic encephalopathy and spinal muscular atrophy type I]. AB - The authors describe the case of a floppy baby with the typical features of early myoclonic encephalopathy, represented by erratic and partial myoclonus of early onset and electroencephalographical suppression-burst pattern. Muscle biopsy made it possible to recognize an important neurogenic pattern, suggesting a severe form of spinal muscular atrophy. The association of these two disorders has never been reported. PMID- 7476750 TI - [Budd-Chiari syndrome in children: case report]. AB - This work aims at describing a particular case of Budd-Chiari syndrome in a ten year-old girl. In our patient, the subjective and objective features of this disease, which is already rare during childhood, are associated with a well assessed polyposis in the blind intestine (intestinum caecum). In our opinion, a possible connection between these two pathological events, a connection unheard of in the literature so far, is an interesting issue to be dealt with in this case report. The follow-up of our patient took place at the Pediatric Hospital of the University of Rome "La Sapienza" and lasted an overall period of 6 months. The girl was hospitalized twice and she underwent a large number of radiological and laboratory tests. The therapy consisted only of medical treatment and support measures. Liver transplant is regarded as the only chance to obtain a full recovery, but it has not yet been performed on our patient. It is by comparing those researches carried out on similar cases with the outcome of our personal study, that we come to the conclusion that, as for our patient, the Budd-Chiari syndrome can be classified as an idiopathic syndrome. Moreover, the fact that this syndrome and the polyposis of caecum occur at the same time in a patient can probably be considered as a coincidence. PMID- 7476751 TI - [Insulin edema in three adolescents with insulin-dependent diabetes mellitus]. AB - Three female patients with a previously poorly controlled Insulin Dependent Diabetes Mellitus (IDDM), without evidence of cardiovascular, hepatic or renal dysfunction, developed generalized edema after a substantial increase in their insulin dosage. Edema resolved in 2-3 weeks, without specific therapy. Our patient's findings met the criteria of diagnosis of insulin edema. Insulin edema during IDDM is an uncommon complication of insulin therapy (1/400) and its pathogenesis is not clarified so far; it is a transient and self-limiting condition. The diagnosis is based on exclusion of all other major causes of edema. PMID- 7476752 TI - [Efficacy and tolerance of azithromycin in respiratory tract infections in children]. AB - This study was carried out to determine the clinical efficacy and the clinical safety of azithromycin in a group of children with acute respiratory tract infections. The study involved 82 children treated with a single daily 10 mg/kg dose of azithromycin for three consecutive days. 7 days later, the overall clinical response was 100% (cure and improvement): bacteriological cure was achieved in 97.5% of the patients treated. Recurrences were never observed. Side effects not requiring interruption of therapy were observed in 3 patients (3.6%). The side effects were gastrointestinal disturbances. In conclusion azithromycin showed a remarkably clinical efficacy for treatment of acute respiratory infections in children. Tolerability and therapeutic compliance were excellent. PMID- 7476753 TI - [Progress and prospectives in the use of hematopoietic stem cells in pediatric oncohematology]. PMID- 7476754 TI - [Acute lymphoblastic leukemia in adolescents]. AB - Age is an important prognostic factor in acute childhood lymphoblastic leukemia (ALL): intermediate age children (1-9 years) show a better outcome than infants (0-1 year) and adolescents (10-15 years). However recent literature data do not agree about adolescents worse prognosis. We tried to contribute to this issue with a retrospective analysis about presenting features and survival of 302 pediatric patients (65 adolescents and 237 children) with non B ALL enrolled on AIEOP protocols at the Departments of Pediatric Haematology-Oncology (University of Turin) from 1976 to 1992. The last follow up was 30.11.94. We found in adolescents, in spite of higher frequency of unfavourable prognostic factors (Hb > 8 g/dl, mediastinal mass, T cell immunophenotype, L2 morphology), an event free survival similar to children (EFS 52% vs 51%). In conclusion in our population we found that age at diagnosis greater than 10 years does not represent an unfavorable prognostic factor. PMID- 7476755 TI - [Neurological manifestations in HIV-infected child]. AB - The authors describe the main epidemiological, pathogenetic and clinical features of HIV-related encephalopathy in 50 pediatric patients (36 females, 24 males), born to HIV-seropositive mothers or infected by contaminated blood, observed in the Institute of Infectious Diseases of Pavia, between January 1984 and December 1994. All the patients (age 1 day-7 years) received a comprehensive evaluation, including subsequent neurodevelopmental assessments, by a multidisciplinary equipe of pediatric infectivologists and neuro-psychiatrists: the follow-up ranges from 6 months to 8.5 years. To evaluate the role of potentially covariates in HIV-vertically infected children, some maternal, gestational and perinatal factors were investigated. The neurodevelopmental assessment was carried out by a standardized protocol: tests were always conducted and evaluated by the same examiner. Seventeen pediatric patients with HIV-related neurologic impairment were observed (34%): 16 cases of encephalopathy (static-stable = 4, plateau = 6, subacute-progressive = 5, uncertain origin = 1) and 1 case of neurotoxoplasmosis. Findings of the current study demonstrate the high frequency of neurological impairment in HIV-infected infant/child, a worse evolution in this kind of patients and the effectiveness of an antiretroviral therapy only if administrated in the early phases of the disease. It was also emphasized the pathogenetic role of some covariates poorly evaluated in previously reported studies. PMID- 7476756 TI - [Cowper's syringocele in children: report on ten cases]. AB - Cystic dilatation of Cowper's gland ducts (Cowper's syringocele) is uncommon in pediatric age. Cowper's syringocele frequently are asymptomatic, sometimes may cause dysuria, pollakiuria and obstructive voiding symptoms. We report ten children with syringocele. Five were observed for recurrent urinary infections, three for obstructive voiding symptoms and two for post-traumatic hematuria and for urinary incontinence. Only three patients with obstructive urethral symptoms were surgically treated. In our opinion careful clinical, radiologic and endoscopic evaluation is necessary in such cases so that unnecessary surgery can be avoided. PMID- 7476757 TI - [Obesity and hypercholesterolemia in a primary school in Rome]. AB - The research in particular the incidence of obesity, hypertension and hyperlipemia was evaluated involved 201 primary school children. The results show an obesity percentage equal to 28%; 23% of the young were hypercholesterolic and 21% had hypertriglyceridemia. Nobody was hypertensioned. The high percentage of obese and hjperlipemic subjects suggest the immediate needs of Courses of alimentary education in schools, as well as to resort to sound daily sporting activity. PMID- 7476758 TI - [Triploidy syndrome: a case report]. AB - Triploidy is a complete extra set of chromosomes and results in a clinically recognizable lethal syndrome with hydatiform placental changes, severe intrauterine growth deficiency, cranial bone abnormalities, eye defects, malformed ears, syndactyly, genital anomalies, congenital heart defects and brain anomalies. Less severe are instances of diploid-tetraploid mixoploidy and patients are more likely to survive. The present report is another contribution to the knowledge of human triploidy: we describe a 69,XXY triploid male occurred to our observation. PMID- 7476759 TI - Shifting balance of power. PMID- 7476760 TI - Forecasting physician workforce requirements in Minnesota. AB - The existing oversupply, specialty imbalance, and geographic maldistribution of clinicians are inconsistent with the goals of health system reform in Minnesota. Health professional workforce projections can help the private and public sectors make informed decisions to assure that the future supply of health professionals reasonably approximates requirements. Appropriate methodologies for projecting national health professional requirements, based on existing HMO staffing patterns, should be applied to Minnesota. In the absence of workforce projections, a variety of assumptions about changes in Minnesota's health care system that will affect workforce supply and requirements can, nonetheless, be made. Limitations on the state's ability to influence the health professional workforce within its borders are noted. PMID- 7476761 TI - Recruitment and retention of family physicians. AB - This study investigates the factors family physicians report as influencing their recruitment and retention. Fifty family physicians who completed their residency training at the Duluth Family Practice Residency received surveys one year and three years after entering practice. Spouses' preferences were most important at the time of recruitment, while children's issues were more influential at the time of the retention survey. The diversity of the higher-ranked variables indicates the multifactorial nature of decisions to settle and maintain practice in a community. Communities and facilities that are recruiting family physicians need to work together. The community's educational system and needs of the spouse must be directly addressed for effective recruitment and retention. Income is important to retention, even though individuals ranked it low as a variable. PMID- 7476762 TI - MMA discusses Medicare with Minnesota delegation. PMID- 7476763 TI - Starting practice in 2005? How will you get GME? PMID- 7476764 TI - Maintain Medicaid safety net, MMA urges. PMID- 7476765 TI - Analysis of AMA Medicare proposal shows savings of $162 billion. PMID- 7476766 TI - Freedom of choice in health care today. Myth or reality? PMID- 7476767 TI - Data privacy under MinnesotaCare. PMID- 7476768 TI - The tension is palpable. PMID- 7476769 TI - One for the money: the financial benefits of EDI. PMID- 7476770 TI - Serious business. PMID- 7476771 TI - Showcasing primary care. PMID- 7476772 TI - [The prognostic significance of the presence of Actinobacillus actinomycetemcomitans in the subgingival plaque of young orthodontic patients]. AB - The aim of present study was to evaluate the prognostic significance of the presence of Actinobacillus actinomycetemcomitans in subgingival plaque from young patients wearing fixed orthodontic appliances. Fifteen orthodontic patients, 3 males and 12 females, aged 10 to 27, systemically healthy, participated in the study. First examination took place in the spring of 1992 (T1) and consisted in the collection of clinical (pocket depth, PD; Gingival Index, GI; Plaque Index, P1I; Gingival Bleeding Index, GBI) and microbiological (Actinobacillus actinomycetemcomitans presence) parameters in 4-8 periodontal sites in each patient. The same clinical and microbiological parameters were collected again in the spring of 1994 (T2). Results showed that the presence of Actinobacillus actinomycetemcomitans in the examined periodontal sites underwent fluctuation, 25% of sites being positive for the sought after bacterium in T2 whereas the percentage of Actinobacillus actinomycetemcomitans positivity in T1 was 47.5% with a statistically significant difference (p < 0.005). On the contrary clinical parameters did not show significant modifications through the study period T1-T2. Differential analysis of clinical parameter modifications in relation to Actinobacillus actinomycetemcomitans positivity in T1 or Actinobacillus actinomycetemcomitans-positivity fluctuations in T1-T2 period did not evidence any significant difference among various microbiological conditions. The authors conclude that the mere presence of Actinobacillus actinomycetemcomitans in subgingival plaque from specific periodontal sites in young orthodontic patients does not seem sufficient to consider those sites at risk for periodontal destruction in the medium-short term. PMID- 7476773 TI - [A polymorphism study of the DNA extracted from dental tissues]. AB - Often teeth are the only items which can be used for personal identification in forensic medicine. In the present work we describe a method to extract and amplify DNA from dental elements ranging from 2 weeks to 5 year from the avulsion. PCR (polymerase chain reaction) was used to amplify VNTR sequences; the alleles products were electrophoresed, visualized by traditional methods and compared to the amplified products obtained from the matching blood sample. Our results give a new and powerful investigative tool for personal identification in the field of forensic odontostomatology, since such a procedure can be successfully applied both to recent and to ancient teeth. PMID- 7476774 TI - [Hypodontia: considerations on orthodontic therapy in agenesis of the permanent upper lateral incisor]. AB - The authors have investigated different clinical cases having in common the agenesis of the permanent upper lateral incisors, to evaluate from a clinical and esthetic point of view the possibilities, results and the limits of the orthodontic treatment. These are nowadays increasingly associated with esthetic dentistry, prosthodontics, implantology and finally with dental transplantation. The patients presented the absence of one or both permanent upper lateral incisors or the agenesis of other teeth, sometimes associated with dental crowding or a not favourable intermaxillary relation. Most of the cases were treated with compensatory extraction of the other normal or hypoplastic permanent upper lateral incisor. The results were satisfactory but the mean treatment time was relatively long (56 months, including the retention phase), according to the difficulty of obtaining a good esthetic result in the smile line. The importance of a correct point of time of the interception for the choice of the most adequate therapy is emphasized. The possibility of correcting occlusal relation by means of the orthopedic treatment of the maxillary structures is limited to the end of the deciduous dentition. The diagnostic criteria for a correct and opportune interception of this anomaly are being stressed on. Finally the esthetic problems arising of the orthodontic space closure and the possible solutions were discussed. PMID- 7476775 TI - [Reconstruction of the oral cavity: the free radial forearm flap versus the free jejunal flap]. AB - The concentration in a restricted area such as the oral cavity of the essential anatomic structures for mastication, deglutition, speech, salivary drainage and respiration makes it indispensable to ensure not only the structural reconstruction of the region but also, and above all, a functional reconstruction of the anatomic unit affected by resection. The use of revascularised flaps has extended both the quantity and quality of reconstructive methods available. In the context of the oral cavity the most widely used flaps are the radial forearm free flap and jejunum free flap. In this paper the authors report their personal experience in a group of 13 patients (6 radial forearm and 7 jejunum) undergoing oral cavity reconstruction using free flap. For each flap the authors describe the microsurgical procedure, the clinical characteristics of the post-operative period, the locoregional complications, the donor site and lastly the long-term clinical, anatomopathological and functional modifications 6-12 months after primary treatment. Moreover, they highlight the varying characteristics of the two flaps and make a critical assessment of the advantages and disadvantages of using one or other method. Lastly, in the light of their experience and a review of international literature, the authors underline the importance of making a careful choice and personalized reconstruction, and finally outline their own criteria of choice. PMID- 7476776 TI - [Hemostasis in patients at high hemorrhagic risk]. AB - Surgical treatment in patients with heavy coagulation disorders needs measures that minimize immediate and post-surgical hemorrhagic risk. Besides the subjects afflicted with constitutional coagulation pathologies such as haemophilia A, B, C, AB, Von Willebrand disease, platelet disorders and others, too many are the patients that need pharmacological therapy to impede the creation of intra vascular coagula. Today, surgical treatment of these patients is possible with the use of human fibrin glue (Tissucol) that permits excellent surgical and post surgical hemostasis without therapy suspension, independently of coagulation disorder. The purpose of this study is a clinical evaluation of a surgical procedure based on the use of human fibrin glue in association or not with suture, hemostatic sponge and lyophilized dura mater in patients with coagulation disorders. In a group of 309 patients in oral anticoagulant therapy have been done a total of 822 operations such as root fragments extraction with mucoperiosteal flap elevation, teeth or roots simple extractions, third molar extractions, cyst removal, apicoectomies. Only patients with PA over 20% have been treated. All the operations have been leaded with minimal trauma; after the extractions, alveolar margins have been regularized to improve flaps adaptation. Hemostatic sponge or dura mater have been used like basis for the flaps and the fibrin glue, that was injected in the residual cavity after surgical suture. In a few cases, for example with little extraction wounds, only fibrin flue without suture or sponge has been used. In all cases and after all operations, the presence or absence of bleeding has been evaluated after 2 hours, 24 hours, 7 days after surgery.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7476777 TI - [Variation, over time, in the pH of the materials used for pulpal-dentinal protection. A new measurement method]. AB - A modern AmpHel pH-meter permits the easy determination of dental materials pH. This new instrument has allowed, analysing seven cements and two varnishes used for pulpodentinal protection, continuously to specify this important parameter's modifications in the long run. The results of this study, conducted for six hours after cement mixing or varnishes application, certified glass ionomer cements' (Ketac-Cem, Fuji-Ionomer II) strong acidity, even higher than well known zinc phosphates' acidity is. A cavity liner's (Tubulitec) pH from an initial value of 6.07 decreased to 5.04 at sixth hour. On the contrary, much lower were the acidity of a modified ZOE (Temrex) and the one of a cavity varnish (Copalite): both showed a physiological pH from the beginning of the test. PMID- 7476778 TI - [Maintenance therapy: an indispensible complement to periodontal treatment and the surgical implant]. AB - The authors analyze the literature of recent years on maintenance therapy both in parodontopathic patients and in subjects undergoing surgical implantation therapy. We divide the work in two parts; the first shows the general aspects of maintenance therapy, the second shows the specific aspects of maintenance therapy in non surgical parodontal treatment, in after surgical parodontal treatment and after implantation treatment. PMID- 7476779 TI - [Granular-cell tumor of the oral cavity (a case report and statistical notes)]. AB - The granular cell tumor is an uncommon benign lesion that may occur anywhere in the body presenting as a single nodular lesion. A case is reported with bilateral palatal lesions. This case, in authors' opinion, is exceptional both for its localization, near palatine foramina, suggesting a neural crest tissue origin, and bilaterality and symmetry pointing out its plurifocal potentiality. PMID- 7476780 TI - [The debate that involves the "dentist" category over the problem of fiscal checks]. PMID- 7476781 TI - [The effect of subgingival irrigation with acetylsalicylate on the polymorphonuclear leukocyte count in periodontal pockets of medium depth]. AB - Considering the role played by arachidonic acid metabolites in the pathogenesis of periodontal disease and the possible beneficial effects exerted by non steroidal anti inflammatory drugs on periodontal damage, the aim of the present study was to assess the capability of a 1% acetylsalicylate solution, delivered by subgingival irrigation, in reducing the number of polymorphonuclear leukocytes (PMN) in periodontal pockets of patients affected by periodontitis. Ten patients systemically healthy, in the age between 36 and 64, participated in the study. After supragingival scaling in each patient test and two control sites were chosen among those exhibiting a pocket depth between 5 and 7 mm. The number of periodontal pocket PMN was evaluated after subgingival washing by the technique of Skapski & Lenher and Gingival Bleeding Index (GBI) score was recorded in each experimental site. Test sites then received subgingival subgingival washing by 0.5 ml of a 1% acetylsalicylate solution three times a week whereas control sites were irrigated by physiologic solution. After treatment the number of PMN and GBI scores from each test and control site was re-evaluated. Results showed that test sites exhibited a decrease in the number of pocket-PMN respect to control sites although not statistically significant (p > 0.5). At contrary GBI score did not modify through the study. In conclusion subgingival irrigation by acetylsalicylate demonstrated to be able to reduce subclinical inflammation in periodontal pockets evaluated by the number of PMN in gingival fluid. PMID- 7476782 TI - [Morphological changes on gold-coated brass endodontic pins]. AB - Scanning electron microscope (SEM) connected with X-ray microanalysis (EDX) was used to study surface alterations of gold-coated brass dental pins (Dentatus), taken out after some years from restored teeth due to necessity for repeating a restorative therapy. The study method led to establish that: --the gold coating of the base material, constituted of copper alloy, absolutely insufficient to prevent corrosion attack; --the observed morphological alterations are the consequence of a localized corrosion attack, produced by anaerobic bacteria, which origin is confirmed by the presence of S on the most attacked points; - finally the corrosive phenomenon does not cause a periapical pathology, but the process is contrary, or rather, it is the pathology that leads to the favourable conditions to create corrosion. PMID- 7476783 TI - [Mandibular movements and the position of the head]. AB - The aim of this study was to evaluate the relation between head position and mandibular movements. The movements of the incisal point were evaluated on a sagittal plane in a group of 4 subjects unaffected by anomalies of the masticatory system. It was observed that there was a marked increase in rotary movement along the hinge axis when the head was extended and vise versa. The author attribute this to interference between the sternocleidomastoid muscles and the mandibular corners (ISM). ISM limits rotary movement, activates mandibular translation and might be the cause of some deviations during opening, malocclusion and ATM disorders. The present study does not establish whether or not ISM plays a functional role. PMID- 7476784 TI - [The malignant transformation of leukoplakia of the oral cavity. A review of the literature and clinical case reports]. AB - After considering the leukoplakia as defined by the WHO, the authors examine the quantitative, qualitative, histopathologic and clinical aspects of leukoplastic lesions. Afterwards, the data concerning the incidence of the malignant evolution of leukoplakia are shown. Data that have been collected and shown during years of study and that are often discordant. For this reason a study on the literature since 1931 is carried out and all the possible causes that could have led to different results are mentioned. Than the authors, among 98 cases of leukoplakia, examine those that turned malignant. In five cases, after the first biopsy, the histopathological report found the presence of microcarcinoma while, in three other cases, in the follow-up, a malignant evolution was observed. This This study points out a percentage of malignant transformation of the leukoplakia equal to 2,94% in agreement with the study of Pindborg and Banoczy. PMID- 7476785 TI - [The therapy of leukoplakia of the oral cavity. A review of the literature]. AB - The authors examine the different therapeutic approaches that can be used to treat the leukoplakia of the oral cavity by examining the literature and doing a critical evaluation. Great importance is attached to methods like CO2 laser therapy, to the treatments with natural or synthetic retinoids, and at last, but not least traditional surgery. These seem to be the most efficient treatments. The data reported seem to confirm the role of surgery (traditional, laser) and the validity of pharmacological therapy with retinoids. On the other hand it is not possible to find long term follow up with bleomycin, selenium and beta interferon, even if at the beginning they seemed to be arouse lively interest. PMID- 7476786 TI - [Amalgam and the toxicological risks of mercury. A review of the argument]. AB - The employment of dental amalgam in the area of conservative dentistry, in spite of representing an excellent therapeutic solution, is today undergoing criticism more frequently than before because of the potential toxicity of mercury regarding hyman health and environmental salubrity. Mercury is a metal having the tendency to evaporate at ambient temperature and it is just the inhalation of its vapor the most common form of exposure to mercury for man. The absorption may also may also occur through food ingestion or accidental swallowing of metal particles coming from dental amalgam fillings. Mercury poisoning usually turns out to be a chronic process which causes its major toxic effects at the expense of digestive, excretory and neurocentral apparatuses. Several authors have studied mercury exposure on the working places, including in their investigation persons operating in the dentistry departments: they found high levels of mercury existing both in tissues and in biological fluids in people affected by exposure, frequently accompanied by symptoms of chronic intoxication. More recent studies, however, do not fully confirm such conclusions and exclude that coming into contact with mercury vapor on the working place may be in connection with an increase of mortality. Exposure to mercury from amalgam for the most part occurs through ingestion of amalgam particles abraded from restored surfaces, inhalation of vapors generated during dental restorative procedures and deglutition of saliva into which corrosion products have dissolved.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7476787 TI - [Odontogenic sinusitis with cutaneous fistulization. A case report]. AB - A 70-year-old male patient presenting with a cutaneous node of the genal region that was first diagnosed as basal cell carcinoma is described. Owing to the following observation, the authors were prompted to a diagnosis of cutaneous odontogenic sinusitis: a) examination of the lesion which emitted pus upon gentle squeezing; b) presence in the vestibular fornix of a large fibrous cord; c) the serious conditions of the mouth and the radiologic results of homolateral sinusitis. The importance of the sinusitis picture, and the necessity to completely remove all bacteria foci prior to surgery on the abdominal aorta the patient was scheduled to undergo, prompted the authors to avulse the cutaneous lesion with tooth extraction, after a "Caldwell-Luc" operation of the homolateral paranasal sinus. Full restitutio ad integrum was achieved. PMID- 7476788 TI - Shifting ground: enteral vs parenteral nutrition in critically ill patients. PMID- 7476789 TI - Clinical applications of enteral nutrition. AB - Clinical and experimental evidence confirms that delivery of nutrients via the gastrointestinal tract reduces septic morbidity in critically injured patients. Early enteral feeding seems to maintain mucosal integrity and to support the gut as an important immunologic organ that may affect other areas of the body. There is increasing evidence to suggest that specific nutrients are especially beneficial in maintaining intestinal host-defense function in times of critical illness and injury. PMID- 7476790 TI - Enteral access. AB - The enteral route is the preferred method of nutrition support in patients with functional gastrointestinal tracts. Many techniques for obtaining enteral access are available, and the decision regarding which one to use depends on several issues, including anticipated duration of support, aspiration risk, and local expertise. Using laparotomy, laparoscopy, fluoroscopy, or endoscopy, tubes can be placed into the stomach, the duodenum, and the jejunum. Nasogastric and nasoenteric tubes are useful for short-term supplementation; however, patients needing support for more than 6 weeks may be better served with a more permanent tube. In this review, specific methods for obtaining enteral access are discussed along with their advantages and disadvantages. PMID- 7476791 TI - pH testing of feeding-tube aspirates to determine placement. AB - Although radiologic confirmation of tube placement remains the "gold standard," there is growing evidence that pH testing of feeding-tube aspirates can reduce (although not totally eliminate) reliance on x-rays used for this purpose. This article describes that evidence and tells how it was obtained. Reducing the number of x-rays needed to confirm tube position can have far-reaching financial benefits. PMID- 7476792 TI - Torsade de pointes and elevated magnesium and calcium requirements associated with intravenous pentamidine. AB - We present a report of markedly elevated calcium and magnesium requirements in an AIDS patient receiving central venous nutrition. These increased requirements were attributed to urinary magnesium losses associated with intravenous pentamidine therapy. The combination of intravenous pentamidine and electrolyte abnormalities resulted in the development of the dysrhythmia, torsade de pointes, in the patient. A review of literature presenting similar cases is included. Nutrition practitioners should be aware of the often prolonged effect of intravenous pentamidine on magnesium and calcium homeostasis so that adequate supplementation will be provided, perhaps preventing the development of life threatening torsade de pointes. PMID- 7476793 TI - Drug-lab interactions: implications for nutrition support. Acetylcysteine interference with urine ketone test. AB - It is important that health care professionals be aware of the potential for medications to interfere with clinical laboratory tests. Medications can cause in vivo effects when the concentration or activity of the analyte is altered before the analysis and therefore the assay result is true and accurate. An in vitro effect occurs when the medication interferes with the assay, and the result is erroneous and cannot be interpreted. This report describes a recently identified case of interference of acetylcysteine with the urine test for ketones and demonstrates the importance of a thorough medication review in evaluating abnormal laboratory tests. PMID- 7476794 TI - Guidelines for nutrition support: guidebook, cookbook, or coupon book? PMID- 7476795 TI - Practice parameters versus outcome measurements. How will prospective and retrospective approaches to quality management fit together? AB - There have been increasing efforts to manage quality and costs in health care. Two tools that are used to do this are practice parameters and outcome measurements. Practice parameters are strategies for patient management to assist clinicians in clinical decision making. Outcome measurements inform clinicians and the public about performance in the delivery of care. Federal and state health care legislation make use of these two concepts as a way to monitor and improve quality and costs, reduce liability, and in some cases, promote competition. Practice guidelines are more limiting to clinicians, but may reduce their liability and help some meet expected clinical outcomes. Outcome criteria alone provide more flexibility to clinicians in deciding how to improve clinical outcome and reduce costs, and enable patients to select care givers on the basis of performance. PMID- 7476796 TI - Nutrition support teams: role in the new health care environment. AB - Many hospitals have strong political leaders who question the need for a nutrition support service. In addition, the pressures of economic and health care reform are forcing administrators to cut unnecessary and unproven, albeit beneficial, services. The challenge for nutrition support services is clear-cut: in order to survive, nutrition support teams must understand the changes in health care and must learn how to adapt to them. This article clarifies the issues and assists nutrition support teams in establishing a new direction. PMID- 7476797 TI - Starting a nutrition support team: short-term pain for long-term gain. AB - This article relates the collective wisdom of five nutrition support professionals who share their experience in starting a nutrition support team. These professionals describe how to navigate the administrative--and political- waters, which are often murky and filled with obstacles. In addition, a hospital administrator discloses what CEO types look for when clinicians propose starting up a nutrition support team in their institutions. The upshot: In this rapidly changing environment, nutrition support professionals would do well to hone their political skills and use them to persuade their administrators that nutrition support is indeed a cost-effective therapy that benefits patients--and that it is best delivered in an interdisciplinary approach. PMID- 7476799 TI - Worksheet for nutrition support services. PMID- 7476798 TI - The nutrition support team: surviving and thriving in an era of reform. AB - Nutrition support teams today are being buffeted by the winds of change in the health care system. With the emphasis shifting from whether to deliver care to how to do so and to how much that care will cost, nutrition support professionals must stay attuned to their institution's needs--and be prepared to adapt to those needs. This article reviews the types of data collection necessary for teams to remain solid as well as offering tips from experts about ways to harness, and even capitalize on, the changes. PMID- 7476800 TI - Nutrition support for lung transplant patients. AB - Lung transplantation has emerged as an established and accepted therapy for patients with endstage pulmonary disease. Very little information has been published about the nutrition management of these patients during the period after transplantation. We conducted a retrospective review of the nutrition support records of 52 adult patients who had undergone lung transplantation at our institution. In addition to patient demographics, data were collected on baseline nutritional status, energy and protein goals, type of specialized nutrition support therapy, length of therapy, and incidence of metabolic complications. More than 50% of the patients receiving lung transplants had a diagnosis of cystic fibrosis. Compared with patients having other diagnoses, this group of patients had a greater incidence of malnutrition. The mean energy goal was 127 +/- 0.07% of basal energy expenditure, and the protein goal was 1.37% +/- 0.25 g/kg/d. All patients received parenteral nutrition therapy, which was begun by postoperative day 2 for more than 70% of the patients. Therapy was short-term (mean, 9 days), and patients then received oral diet. The most common metabolic complications were azotemia, hyperglycemia, and hypomagnesemia. Our experience should provide assistance to other health care professionals who are involved in the nutrition and metabolic management of patients undergoing lung transplantation. PMID- 7476801 TI - Clinical comparison of three methods to determine resting energy expenditure. AB - An accurate and reliable method of determining resting energy expenditure in the critical care setting is crucial because inadequate caloric intake and excessive caloric intake are both associated with a number of complications. Energy requirements were determined by three different methods in a group of 25 post cardiac surgery patients in the intensive care unit. Patients were intubated and had a thermodilution pulmonary artery catheter in place. The first method measured resting energy expenditure by indirect calorimetry. For the second method, the results of blood gases drawn at the same time that indirect calorimetry was measured were used in the Fick equation. In the third method, a registered dietitian assessed each patient by using the Harris-Benedict equation with stress factor modification. Indirect calorimetry was considered the standard with which the other two methods were compared. The results showed that, compared with calorimetry, both the Fick equation and the Harris-Benedict equation underestimated resting energy needs. Statistically, only the difference between the Fick equation method and indirect calorimetry was significant. Clinically, however, both methods seem to have applicability. PMID- 7476802 TI - Possible folate deficiency with postsurgical infection. AB - A 51-year-old obese woman was transferred to our hospital for management of a complicated laparoscopic cholecystectomy accompanied by fever and malaise. A liver abscess was discovered. On postoperative day 52 it was noted that the patient's tongue was magenta and sore and that she had altered taste, pallor, severe weight loss, diarrhea, and poor appetite and mood. A dermatology consult suggested that her tongue abnormalities were most likely nutrition-related, and a hematologic blood smear was suggestive of folate deficiency. Premorbidly, the patient had consumed a diet chronically low in folate and had received estrogen therapy for 15 years. Throughout the patient's hospitalization, she had bouts of fever and received numerous antibiotics. Within a month of initiating nutrition support and vitamin/mineral supplements, her nutritional status improved dramatically. This patient's poor diet, long-term estrogen use, surgical stress, infection, and lack of early nutrition support may have contributed to her possible folate deficiency and her prolonged hospital stay. PMID- 7476803 TI - Third international conference on nutrition and HIV/AIDS focuses on the issues. PMID- 7476804 TI - Frustration or opportunity? The impact of managed care on emergency psychiatry. PMID- 7476805 TI - Academic issues in emergency psychiatry. PMID- 7476806 TI - The spectrum of mobile outreach and its role in the emergency service. PMID- 7476807 TI - Crisis residential treatment: an alternative to hospitalization. PMID- 7476808 TI - An integrated approach to emergency psychiatric care. AB - Increased demands for psychiatric emergency services led to a crisis in New York City emergency rooms. The New York State Comprehensive Psychiatric Emergency Program was developed as a systemic response to the crisis. The model includes four integrated components designed to enhance the entire mental health system. PMID- 7476809 TI - Emergency psychiatric evaluation of "organic" mental disorders. AB - The emergency psychiatrist must first differentiate abnormal behavior with a medical etiology from that caused by so-called functional processes. Delays in diagnosis can be catastrophic and make a compelling argument for physician involvement in emergency mental health care. PMID- 7476810 TI - Crisis hospitalization in a psychiatric emergency service. AB - Crisis hospitalization has both an affirmative rationale in the community movement and a secondary rationale of cost containment. Brief hospitalization within the psychiatric emergency service offers intensive treatment and a rapid return to the community for the patient and is cost-effective for the mental health system. PMID- 7476811 TI - Diagnosing substance abuse in psychiatric emergency service patients. PMID- 7476812 TI - Psychiatric emergencies in children and adolescents. PMID- 7476813 TI - Psychological treatments in the psychiatric emergency service. PMID- 7476814 TI - Emergency pharmacotherapy: the evolving role of medications in the emergency department. PMID- 7476815 TI - Management of aneurysmal subarachnoid hemorrhage. AB - Treatment of ischemic deficits caused by vasospasm relies on enhancing cardiac output, inducing arterial hypertension, and expanding the intravascular volume in an attempt to improve CBF. Different treatment protocols exist from institution to institution to achieve these goals. The role of calcium-channel blockers now is well established. The newest focus on prevention of vasospasm includes tPA and a variety of anti-inflammatory drugs and potential neuroprotective drugs under research. Endovascular therapy for vasospasm has an increasing role in treating patients who are unable to tolerate induced hypertension or aggressive volume augmentation. We will return to our index case of the 63-year-old woman with SAH caused by an ACoA aneurysm to review some major management issues. After placing a ventriculostomy and slowly lowering ICP, the patient became alert and was fully oriented. She had aneurysm surgery on hospital day 2, with an uncomplicated immediate postoperative course. A Swan-Ganz catheter, placed for intraoperative monitoring, was kept in place and she was hydrated with 125 mL/hour of normal saline, achieving a PAWP of 10 to 16 mm Hg. Her mean arterial blood pressure without pharmacologic intervention was 95 to 110 mm Hg. She had continued clinical improvement with resolution of her left hemiparesis. On hospital day 5, her ventriculostomy was clamped because cerebrospinal fluid drainage was minimal. The following morning, the patient was arousable only to deep pain and her left side was flaccid. An emergent CT scan demonstrated no new hemorrhage, no increase in ventricular size, and no infarct. Vasospasm was considered the most likely cause. Hypertensive therapy was about to be initiated with a phenylephrine drip, but within an hour she was fully alert and moving all extremities equally. A search for other potential causes of neurologic decline was undertaken and revealed a phenytoin level of 5.5. It was thought that the patient most likely had had a seizure and that her clinical deterioration represented a postictal state. She received a bolus infusion of phenytoin. On hospital day 7, the patient became confused, insisting that her nurse was her son and ordering him out of her "apartment." Lower extremity weakness was detected. CT scan was unchanged. Phenylephrine was started but she developed precordial lead ST elevation and elevated cardiac enzymes. Topical nitrate therapy was initiated and phenylephrine was discontinued. The patient underwent emergent cerebral angiography, which demonstrated moderate to severe bilateral ACA spasm and moderate right MCA spasm.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 7476816 TI - Malignant cerebral edema and intracranial hypertension. AB - Cerebral edema and intracranial hypertension occur frequently in neurologic patients. Proper understanding of the pathophysiology of each entity allows prompt recognition and rational therapeutic goals, allowing for better neurologic outcome in many disease states. The recognition of cerebral edema as a distinct entity allows the clinician to treat focal pressure gradients in the brain separately from more diffuse intracranial pressure elevations, appreciating the benefits and pitfalls of directed therapies for each process. The treatment of many of the disorders that cause cerebral edema and intracranial hypertension is heuristic, challenging the managing physician's thorough understanding of cerebral hemodynamics and his or her ability to encounter the human aspects of determining appropriate levels of care for individual patients. PMID- 7476817 TI - Neurocritical care for acute ischemic stroke. AB - Therapy for acute ischemic stroke includes general and specific measures. General management such as respiratory and cardiac care, fluid and electrolyte management, blood pressure control, and treatment of increased ICP is the basis of all stroke treatment. Specific treatment is directed to recanalization of the occluded vessel, control of life-threatening brain edema, and protection of neuronal tissue against the toxic effects of ischemia. Although controlled studies are still lacking, application of the proposed general interventions provides a high-quality standard in patients with acute stroke. Among specific therapies, medical thrombolysis has demonstrated instances of significant and sustained neurologic improvement. Several multicenter trials are underway to confirm these preliminary results. In addition to thrombolysis, cell-protecting drugs hold promise for the future. Presumably, a combination of thrombolytic and cell-protecting agents will be the treatment of choice in early stroke in the future. PMID- 7476818 TI - Status epilepticus. AB - Status epilepticus is defined as a condition characterized by epileptic seizure prolonged or repeated so as to produce a lasting epileptic state. It is estimated that status epilepticus occurs in 50,000 to 60,000 individuals in the United States, with one third to one half of episodes occurring in patients with established epilepsy. PMID- 7476819 TI - Neurologic evaluation of the patient with acute bacterial meningitis. AB - Major advances in our understanding of the pathophysiology of bacterial meningitis have been made in the past decade. It is likely that interventional strategies that mediate the effects of vasoactive metabolites and neuronal and glial toxins will improve the outcome of patients with meningitis as well as other neurologic disorders. Of critical importance, as demonstrated in the case history, is the realization that many of the serious complications of meningitis occur very early in the course of the disease. If new treatment strategies are to be effective, they should be started as soon as possible. Emerging technologies such as proton magnetic resonance spectroscopy may be of benefit in helping physicians decide which patients require treatment. PMID- 7476820 TI - Neurophysiologic monitoring in the neuroscience intensive care unit. AB - Neurologically critically ill patients, more often than others, are unable to communicate and, for a crucial period of time, have the vital functions of their brains hidden in the "black box" of the cranial vault behind a curtain of ambiguity and immobility. Customarily--and naively--we have relied upon beside clinical observations to pierce these barriers. Recent insights lead us to conclude that these "neurochecks" no longer suffice. This article has surveyed four major monitoring systems relied upon by neurointensivists to evaluate the pathophysiology of their patients. Of these, ICPM has the longest clinical track record. It provides a quantitative measure of the brain's capacity to withstand ICP and helps us monitor interventions to reduce it. To utilize this information intelligently requires an understanding of the principles of ICC, CPP, ICP wave morphology, and the hardware available. NICU-CEEG is a more recent introduction but, in principle, it transfers from the laboratory and operating suite to the ICU bedside, established correlations among electrophysiology, CBF, and CM. Digital EEG has allowed us to overcome significant logistical barriers and made NICU-CEEG a practical ICU tool. Early but impressive data suggest that NICU-CEEG has a significant clinical impact in patients with ACI, uncontrolled seizures, or coma. It also has revealed that NICU patients have a surprisingly high incidence of NCS, which may adversely affect their outcome. TCD has contributed greatly to diagnosis and management of SAH vasospasm. It also can be applied with benefit to patients with increased ICP, and has promising value in patients with ACI. It may prove beneficial in monitoring unstable cerebral embolization. Several bedside methods for monitoring CBF are available, but they require refinement to become true monitoring systems. These methods have revealed clinically important insights in patients with head trauma, SAH vasospasm, and ACI. Methods for directly monitoring CM and CMRo2 are improving our understanding of the brain's responses to injury, and becoming increasingly relevant to bedside management. SjvO2 can detect cerebral ischemia caused by overzealous hyperventilation and accelerated ICP. ICO holds promise as a noninvasive transcranial method for assessing Scvo2. We soon may see a scalp array of such detectors, similar to an EEG "montage," that allows us to assess multiregional Scvo2. To be useful, a clinical method should raise questions for further investigation. If the neurophysiologic monitoring systems described here provide us with some answers and lead us to ask useful new questions, they will prove their benefit to our patients. PMID- 7476821 TI - Ventilatory management in critical neurologic illness. AB - Evolution and refinement of airway and ventilatory management has taken place in neurologic critical care. Applications for ventilatory management have been extended to virtually all acute life-threatening illnesses found in neurologic critical care units today. This article focuses on the airway and ventilatory management of patients with acute raised intracranial pressure, cervical spine injuries, and neuromuscular ventilatory failure. PMID- 7476822 TI - Neurologic complications of systemic critical illness. AB - Neurologic complications of systemic illnesses may affect any part of the nervous system in various conditions. In many systemic disorders, however, an acute confusional state is the first manifestation. It is urgent to make a definitive diagnosis and begin specific and supportive therapy to lessen mortality and morbidity. A case of severe septic illness with central and peripheral nervous system complications is reviewed. Septic encephalopathy and critical illness polyneuropathy are common conditions in the general ICU. The neuropathy often is the underlying cause for failure to wean from the ventilator. PMID- 7476823 TI - Critical neurologic illness in the immunocompromised patient. AB - Immunocompromised patients are vulnerable to a host of neurologic problems that may complicate their clinical course and necessitate intensive care. Atypical and opportunistic infections represent the primary concern in such patients and can cause a variety of neurologic complications. In addition, these patients are susceptible to neurologic dysfunction induced by drugs or underlying disease states. This article emphasizes the diagnostic approach to selected clinical syndromes in the immunocompromised patient, such as encephalopathy, seizures, and stroke. PMID- 7476824 TI - Neurologic intensive care unit management and economic issues. AB - This article opens with a case study and shows the complex nature of neurologic intensive care patients. After an historical perspective, the administrative organization and collaborative practice are discussed. The author also addresses the impact of outcome research and managed care on the current and future practice of the neurologic intensive care unit. PMID- 7476825 TI - 1995-6 Membership directory of the Missouri State Medical Association. PMID- 7476827 TI - Survival of the fittest. PMID- 7476826 TI - Hormones and highways: a dangerous mixture for teenage drivers. PMID- 7476828 TI - The answer is, "no! chess for fun". PMID- 7476829 TI - Reinventing the house of medicine: the study of the federation. PMID- 7476830 TI - Otolaryngology quiz #7. Squamous cell carcinoma. PMID- 7476831 TI - Tax-exempt integrated delivery systems: Part 2, Stark I and Stark II. PMID- 7476832 TI - Can Missouri health care providers be held liable under strict product liability law? PMID- 7476833 TI - MSMA women physicians leading change. PMID- 7476834 TI - Retrospective analysis of the utilization of right-sided cardiac catheterization in association with left-sided cardiac catheterization in patients with coronary artery disease. PMID- 7476835 TI - Post tonsillectomy hemorrhage: a ten-year retrospective study. AB - Tonsillectomy remains one of the most common surgical procedures performed in the world. One of the most significant complications is postoperative hemorrhage. Though most bleeding often can be controlled with local anesthesia in an office or emergency room, some patients require return to the operating room for general anesthesia to control the hemorrhage. A ten-year retrospective study was done to review patients who required return to the operating room for general anesthesia for control of hemorrhage. Variables investigated include age, gender, time of surgery, onset of hemorrhage, season of the year, and the amount of blood loss noted during initial surgery. Seven of the 756 patients (.93%) required a second general anesthesia to control hemorrhage. Seasonal and gender predilictions are noted in this study. Recent trends in tonsillectomy are reviewed. Postoperative hemorrhage creates anxiety, prolongs morbidity, and increases health care cost. Limiting this complication is clearly in the best interest of both patient and surgeon. PMID- 7476836 TI - Are serum levels of vancomycin useful in the first week of therapy? AB - Controversy exists regarding the need to monitor serum concentrations of vancomycin with some investigators recommending measurement of peak and trough concentrations in the first week of therapy and regularly thereafter, whereas others contend that empiric dosing produces safe and effective drug concentrations so that testing is unnecessary. Since vancomycin concentrations are measured, routinely in our hospital in the first week of therapy, we conducted a 12 month study to assess their clinical value in patients who were treated when gram positive cocci were detected in blood culture smears. One hundred-five patients had gram positive cocci on blood culture smears. These bacteria were pathogens in 15 patients with Staphylococcus aureus and in 18 with coagulase negative staphylococci based on microbiologic criteria and a chart review confirming their clinical significance. Ten patients with S. aureus and 8 patients with coagulase negative staphylococci that were pathogens and 10 patients with coagulase negative staphylococci that were contaminants were treated with vancomycin. Serum peak and trough concentrations of vancomycin obtained within the first 5 days of therapy in these 28 patients were 14 to 40 micrograms/ml and 4.8 to 20 micrograms/ml. These concentrations were much above the MIC's of the microorganisms (< 4 micrograms/ml). Five patients had increases of serum creatinine of more than 0.6 mg% and in each patient the increases were attributable to other causes-shock, heart failure, and preexisting renal failure. Fifty five peak and trough concentrations 19 of which were drawn in patients with contaminated cultures were measured at a cost of $2,475.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7476837 TI - Air-bag-associated fatal injuries to infants and children riding in front passenger seats--United States. AB - Air bags, when used as a supplement to safety belts, effectively prevent deaths and serious injuries in frontal motor-vehicle crashes. Air bags are standard equipment in most new cars; federal safety standards require that all new passenger cars and light trucks be equipped with both driver- and passenger-side air bags by 1999. The safety of air bags is well documented, and air bags have saved an estimated 900 lives since the late 1980s (1); however, special precautions are needed to safely transport children in vehicles equipped with air bags. Reports of eight deaths of child passengers in crashes involving air-bag deployment are of special concern because they involved low-speed crashes that the children otherwise might have survived. This report summarizes three of these eight cases (2). PMID- 7476838 TI - First 500,000 AIDS cases--United States, 1995. AB - As of October 31, 1995, a total of 501,310 persons with acquired immunodeficiency syndrome (AIDS) have been reported to CDC by state and territorial health departments; 311,381 (62%) had been reported as having died. The AIDS surveillance case definition was substantially expanded in late 1987 and again in 1993 to reflect increased knowledge of the natural history of human immunodeficiency virus (HIV) and to remain consistent with the clinical management of HIV disease (1,2). This report presents rates of reported AIDS cases for 1994 and describes the temporal changes in the characteristics of persons reported with AIDS during three periods corresponding to changes in the AIDS case definition--1981-1987, 1988-1992, and 1993-October 1995--and how this information can be used to plan local, state, and national prevention programs. PMID- 7476839 TI - Deaths associated with a purported aphrodisiac--New York City, February 1993-May 1995. AB - During February 1993-May 1995, the New York City Poison Control Center (NYCPCC) was informed about onset of illness in five previously healthy men after they ingested a substance marketed as a topical aphrodisiac; four of the men died. These cases were investigated by the New York City Department of Health, the New York City Department of Environmental Protection, and the Food and Drug Administration (FDA). Four cases were referred to the NYCPCC and one case to the New York City medical examiner's office. The decedents died from cardiac dysrhythmias, and all five patients had measurable levels of digoxin* detected in their serum. Digoxin had not been prescribed for therapeutic purposes for any of these patients, and none had medical conditions associated with endogenous digoxin-like immunoreactive substances. The purported aphrodisiac contains bufadienolides, naturally occurring cardioactive steroids that have digoxin-like effects. This report describes three of the five case reports, summarizes the investigations of the five cases, and underscores the health risks associated with inappropriate use of preparations containing digoxin-like substances. PMID- 7476840 TI - Update: influenza activity--United States and England, 1995-96 season. AB - In cooperation with the World Health Organization (WHO), its collaborating laboratories, and state and local health departments, CDC conducts surveillance to monitor influenza activity and to detect antigenic changes in the circulating strains of influenza viruses. This report summarizes influenza surveillance activities in the United States and England from September 17 through November 11, 1995. PMID- 7476841 TI - Update: alcohol-related traffic crashes and fatalities among youth and young adults--United States, 1982-1994. AB - Approximately one third of deaths among persons aged 15-24 years result from motor-vehicle crashes (1). Although alcohol use increases the risk for motor vehicle crashes for all drivers, for young drivers the risk begins to increase at very low blood alcohol concentrations (BACs) (2). In addition, in young persons who drive after drinking, the relative risk for crash involvement is greater at all BACs than for older drivers who drink (3). This report is based on data from the Fatal Accident Reporting System of the National Highway Traffic Safety Administration (NHTSA) and describes trends in alcohol involvement among drivers in fatal traffic crashes and trends in all alcohol-related traffic fatalities (ARTFs) in the United States from 1982 through 1994 among youth and young adults. PMID- 7476842 TI - Progress toward global eradication of dracunculiasis. AB - The plan for the global eradication of dracunculiasis (i.e., Guinea worm disease) was developed in October 1980, when dracunculiasis was known to be endemic in 16 African countries, India, and Pakistan. In 1991, the World Health Assembly adopted a resolution to eradicate dracunculiasis by the end of 1995. This report summarizes the substantial progress toward eradication of dracunculiasis since the beginning of this effort. PMID- 7476843 TI - Increasing morbidity and mortality associated with abuse of methamphetamine- United States, 1991-1994. AB - Methamphetamine (also known as "speed," "crystal," "crank," "go," and "ice") is the most widely illegally manufactured, distributed, and abused type of amphetamine, a class of stimulant drugs. An estimated 4 million persons in the United States have abused methamphetamine at least once (1). Information from several sources--including medical examiners, hospital emergency departments (EDs), substance-abuse-treatment facilities, and community epidemiologists- suggests a recent increase in morbidity and mortality associated with abuse of methamphetamine in the United States, primarily in the West but also in the South and Midwest. To characterize trends in methamphetamine-associated morbidity and mortality during 1991-1994, the Substance Abuse and Mental Health Services Administration (SAMHSA) compiled and analyzed data from the Drug Abuse Warning Network (DAWN) and the Treatment Episode Data Set (TEDS). This report summarizes the results of these analyses. PMID- 7476844 TI - Alcohol involvement in fatal motor-vehicle crashes--United States, 1993-1994. PMID- 7476845 TI - Wound botulism--California, 1995. AB - During January-November 1995, a total of 19 laboratory-confirmed cases of wound botulism were reported to the California Department of Health Services (CDHS); of these, 13 had occurred since August. Since 1990, the number of wound botulism cases reported annually in California has increased steadily (one case in 1990, two in 1991, three in 1992, four in 1993, and 11 in 1994). All cases except one since 1991 have occurred in injecting-drug users, and many involved subcutaneous injection or "skin popping" of black tar heroin. This report summarizes the findings of the investigation of two cases. PMID- 7476846 TI - Unexplained severe illness possibly associated with consumption of Kombucha tea- Iowa, 1995. AB - Kombucha tea is a popular health beverage made by incubating the Kombucha mushroom in sweet black tea. Although advocates of Kombucha tea have attributed many therapeutic effects to the drink (1-3), its beneficial and/or adverse effects have not been determined scientifically. During April 1995, cases of unexplained severe illness (including one death) occurred in two persons in a rural town in northwestern Iowa who had been drinking Kombucha tea daily for approximately 2 months. Based on the findings of a preliminary investigation by the Iowa Department of Public Health (IDPH), on April 10 IDPH issued a news release recommending that persons refrain from drinking Kombucha tea until the role of the tea in the two cases of illness had been evaluated fully. This report summarizes the investigation of these cases by the IDPH, CDC, and the Food and Drug Administration (FDA). PMID- 7476847 TI - Update: respiratory syncytial virus activity--United States, 1995-96 season. AB - Respiratory syncytial virus (RSV), a common cause of winter outbreaks of acute respiratory disease, is associated each year with an estimated 90,000 hospitalizations and 4500 deaths from lower respiratory tract disease in both infants and young children in the United States (1). Outbreaks occur annually throughout the United States, and community activity usually peaks within 1 month of the national peak in January or February (Figure 1) (2). RSV activity in the United States is monitored by the National Respiratory and Enteric Virus Surveillance System (NREVSS), a voluntary, laboratory-based system. This report presents provisional surveillance results from the NREVSS for RSV during July 1 December 1, 1995, and summarizes trends in RSV from July 1990 through June 1995. PMID- 7476848 TI - State laws on tobacco control--United States, 1995. AB - PROBLEM/CONDITION: State laws on smoke-free indoor air, youth access to tobacco products, advertising of tobacco products, and excise taxes on tobacco products are summarized. REPORTING PERIOD COVERED: Legislation effective through June 30, 1995. DESCRIPTION OF SYSTEM: CDC and the National Cancer Institute (NCI) identified state laws addressing tobacco control by using LEXIS, which is an on line legal research data base, and NCI's State Cancer Legislative Database (SCLD), which is a data base of legislation. CDC and NCI conducted detailed analyses of the content of the laws to identify specific provisions. RESULTS: CDC and NCI identified 1,238 state laws that address tobacco-control-related issues. Most laws either enact restrictions or strengthen current legislation that restricts tobacco use, sales to minors, or advertising; however, some laws preempt stronger measures by local ordinances. At the state level, forty-six states and Washington, DC require smoke-free indoor air to some degree or in some public places. All states prohibit the sale and distribution of tobacco products to minors, but only nine states restrict advertising of tobacco products. All states tax cigarettes (average excise tax is 31.5 cents per pack); 42 states also tax chewing tobacco and snuff. INTERPRETATION: State laws addressing tobacco control vary in relation to restrictiveness, enforcement and penalties, preemptions, and exceptions. ACTIONS TAKEN: The tables summarizing these laws are available through CDC's State Tobacco Activities Tracking and Evaluation (STATE) system and through NCI's SCLD. This information can be used by policy makers at the state and local levels to plan and implement initiatives on youth access to tobacco products and on the use, promotion, advertising, and taxation of tobacco products. PMID- 7476850 TI - Role of HU proteins in forming and constraining supercoils of chromosomal DNA in Escherichia coli. AB - Induction of supercoiling in plasmid DNA by HU heterotypic and homotypic dimers, a mutant HU-2 (HupAN12), HBs and HB1 proteins with different DNA-binding affinities was investigated in vitro. The abilities of these proteins to induce supercoiling in DNA correlated with their affinities for DNA. Stoichiometrical analysis of HU heterodimers bound to DNA in the complex restraining the negative torsional tension of DNA showed that 12-13 dimers account for a single superhelical turn. The number of supercoils in the plasmid in vivo decreased on inhibition of DNA gyrase with coumermycin, reaching a steady-state level that indicated the existence of a compartment of restrained supercoils. The size of the restrained compartment was reduced in the absence of HU, indicating the participation of HU in constituting this fraction, and was larger on overproduction of HU-2 in the cells. An increased level of DNA gyrase, expressed from a plasmid carrying both gyr genes, in the cells did not compensate for the deficit of the restrained supercoils caused by HU deficiency, indicating seeming distinct and unrelated action of HU and DNA gyrase in introducing and constraining supercoiling of intracellular DNA. PMID- 7476849 TI - A maize DNA-binding factor with a bZIP motif is induced by low temperature. AB - We have isolated a low temperature-induced maize gene, mlip15, via cross hybridization using rice lip19. The longest cDNA isolated comprised 1179 bp and coded for a 135 amino acid bZIP (basic region/leucine zipper) protein. The gene showed 61.4% and 68.9% identity with the rice gene at the DNA and amino acid sequence levels, respectively, and is distinct from other maize genes that code for bZIP proteins. The level of mlip15 transcript was positively regulated by low temperature in the same way as the lip19 transcript. The levels of the transcript were also strongly increased by salt stress and exogenous abscisic acid, and slightly increased by anaerobiosis, but were not affected by heat shock and drought. The mLIP15 protein and truncated derivatives, produced in rabbit reticulocyte lysates or in an Escherichia coli expression system, were able to bind to a fragment of the wheat histone H3 gene promoter. This binding was diminished by addition of a molar excess of the hexamer sequence 5'-ACGTCA-3' found in the promoter and of the G-box-like sequence, but not by the addition of the ocs sequence or a mutated hexamer sequence. The factor bound to a promoter region of the maize Adh1 gene, expression of which is also induced by low temperature. These results lead to the conclusion that mlip15 is a strong candidate for a low temperature-induced transcription factor in maize. PMID- 7476852 TI - Molecular analysis and mapping of two genes encoding maize glutathione S transferases (GST I and GST II). AB - Maize glutathione S-transferase (GST) isozymes are encoded by a gene family comprising at least five genes, three of which (Gst I, II and III) have recently been isolated and sequenced. The enzymes are active as homo or heterodimers and exhibit intraspecific polymorphism including a "null" variant for the two major isoforms expressed in roots. Northern blot analyses performed on total root RNA from "null" and "plus" genotypes, using Gst I- and Gst II-specific probes, indicated that the Gst I gene controls the expression of the two major GST isoforms expressed in roots. Gst I and Gst II were mapped by RFLP analysis using an F2 population of 149 individuals previously characterized. Gst I was localized on the long arm of chromosome 8, while two putative Gst II loci were mapped to chromosome 8 (70 cM from Gst I) and 10, respectively. PMID- 7476851 TI - The binding motifs for Ac transposase are absolutely required for excision of Ds1 in maize. AB - A reverse genetic system for studying excision of the transposable element Ds1 in maize plants has been established previously. In this system, the Ds1 element, as part of the genome of maize streak virus (MSV), is introduced into maize plants via agroinfection. In the presence of the Ac element, excision of Ds1 from the MSV genome results in the appearance of viral symptoms on the maize plants. Here, we used this system to study DNA sequences required in cis for excision of Ds1. The Ds1 element contains the Ac transposase binding motif AAACGG in only one of its subterminal regions (defined here as the 5' subterminal region). We showed that mutation of these motifs abolished completely the excision capacity of Ds1. This is the first direct demonstration that the transposase binding motifs are essential for excision. Mutagenesis with oligonucleotide insertions in the other (3') subterminal region resulted in elements with either a reduced or an increased excision efficiency, indicating that this subterminal region also has an important function. PMID- 7476853 TI - Involvement of two different urf-s related mitochondrial sequences in the molecular evolution of the CMS-specific S-Pcf locus in petunia. AB - In petunia, a mitochondrial (mt) locus, S-Pcf, has been found to be strongly associated with cytoplasmic male sterility (CMS). The S-Pcf locus consists of three open reading frames (ORF) that are co-transcribed. The first ORF, Pcf, contains parts of the atp9 and coxII genes and an unidentified reading frame, urf s. The second and third ORFs contain NADH dehydrogenase subunit 3 (nad3) and ribosomal protein S12 (rps12) sequences, respectively. The nad3 and rps12 sequences included in the S-Pcf locus are identical to the corresponding sequences on the mt genome of fertile petunia. In both CMS and fertile petunia, only a single copy of nad3 and rps12 had been detected on the physical map of the main mt genome. The origin of the urf-s sequence and the molecular events leading to the formation of the chimeric S-Pcf locus are not known. This paper presents evidence indicating that two different mt sequences, related to urf-s and found in fertile petunia lines (orf-h and Rf-1), might have been involved in the molecular evolution of the S-Pcf locus. Southern analysis of mtDNA derived from both fertile and sterile petunia plants suggests that one of these urf-s related sequences (showing 100% homology to urf-s and termed orf-h) is located on a sublimon. An additional, low-homology urf-s related sequence (Rf-1) is shown to be located on the main mt genome 5' to the nad3 gene. It is, thus, suggested that the sequence of events leading to the generation of the S-Pcf locus might have involved introduction of the orf-h sequence, via homologous recombination, into the main mt genome 5' to nad3 at the region where the Rf-1 sequence is located. PMID- 7476854 TI - Tagging pathogenicity genes in Ustilago maydis by restriction enzyme-mediated integration (REMI). AB - In the maize pathogenic fungus Ustilago maydis integration of transforming DNA at homologous or heterologous sites is often accompanied by duplications of the DNA. We show that it is possible to generate single-copy integration events with high efficiency by restriction enzyme-mediated integration (REMI). In about 50% of cases, a plasmid that contains a single BamHI site is integrated at chromosomal BamH1 sites, if BamHI is added to the transformation mixtures. In the other cases it appears that integration events have also occurred preferentially at BamHI sites, but without restoration of the recognition sites. Using REMI we have generated approximately 1000 insertion mutants. Pathogenicity tests demonstrated that about 1-2% of these mutants were unable to induce symptoms when tested in planta. For two of the mutants we have shown that the phenotype is linked to the insertion event. PMID- 7476855 TI - Genetic and physical characterization of the LR1 leaf rust resistance locus in wheat (Triticum aestivum L.). AB - The objective of this study was to characterize the leaf rust resistance locus Lr1 in wheat. Restriction fragment length polymorphism (RFLP) analysis was performed on the resistant line Lr1/6* Thatcher and the susceptible varieties Thatcher and Frisal, as well as on the segregating F2 populations. Seventeen out of 37 RFLP probes mapping to group 5 chromosomes showed polymorphism between Lr1/6* Thatcher and Frisal, whereas 11 probes were polymorphic between the near isogenic lines (NILs) Lr1/6* Thatcher and Thatcher. Three of these probes were linked to the resistance gene in the segregating F2 populations. One probe (pTAG621) showed very tight linkage to Lr1 and mapped to a single-copy region on chromosome 5D. The map location of pTAG621 at the end of the long arm of chromosome 5D was confirmed by the absence of the band in the nulli-tetrasomic line N5DT5B of Chinese Spring and a set of deletion lines of Chinese Spring lacking the distal part of 5DL. Twenty-seven breeding lines containing the Lr1 resistance gene in different genetic backgrounds showed the same band as Lr1/6* Thatcher when hybridized with pTAG621. The RFLP marker was converted to a sequence-tagged-site marker using polymerase chain reaction (PCR) amplification. Sequencing of the specific fragment amplified from both NILs revealed point mutations as well as small insertion/deletion events. These were used to design primers that allowed amplification of a specific product only from the resistant line Lr1/6* Thatcher. This STS, specific for the Lr1 resistance gene, will allow efficient selection for the disease resistance gene in wheat breeding programmes. In addition, the identification of a D-genome-specific probe tightly linked to Lr1 should ultimately provide the basis for positional cloning of the gene. PMID- 7476856 TI - Cloning of the Helicobacter pylori recA gene and functional characterization of its product. AB - The RecA protein is a key enzyme involved in DNA recombination in bacteria. Using a polymerase chain reaction (PCR) amplification we cloned a recA homolog from Helicobacter pylori. The gene revealed an open reading frame (ORF) encoding a putative protein of 37.6 kDa showing closest homology to the Campylobacter jejuni RecA (75.5% identity). A putative ribosome binding site and a near-consensus sigma 70 promoter sequence was found upstream of recA. A second ORF, encoding a putative protein with N-terminal sequence homology to prokaryotic and eukaryotic enolases, is located directly downstream of recA. Compared to the wild-type strains, isogenic H. pylori recA deletion mutants of strains 69A and NCTC11637 displayed increased sensitivity to ultraviolet light and abolished general homologous recombination. The recombinant H. pylori RecA protein produced in Escherichia coli strain GC6 (recA-) was 38 kDa in size but inactive in DNA repair, whereas the corresponding protein in H. pylori 69A migrated at the greater apparent molecular weight of approx. 40 kDa in SDS-polyacrylamide gels. However, complementation of the H. pylori mutant using the cloned recA gene on a shuttle vector resulted in a RecA protein of the original size and fully restored the general functions of the enzyme. These data can be best explained by a modification of RecA in H. pylori which is crucial for its function. The potential modification seems not to occur when the protein is produced in E. coli, giving rise to a smaller but inactive protein. PMID- 7476857 TI - Developmental and tissue-specific regulation of the gene for the wheat basic/leucine zipper protein HBP-1a(17) in transgenic Arabidopsis plants. AB - Wheat basic/leucine zipper protein HBP-1a(17) binds in vitro specifically to ACGT motif-containing cis-acting elements, such as the type I element of plant histone promoters and the G-box of hormone- and light-inducible promoters. To address the in vivo function of HBP-1a(17), we isolated and structurally analyzed the HBP 1a(17) gene and examined its expression in transgenic Arabidopsis plants. The HBP 1a(17) gene is composed of 14 exons; the basic region and leucine zipper are encoded by separate small exons, as is the case for other bZIP protein genes. The G-box of the HBP-1a(17) promoter bound specifically to HBP-1a(17) and its related HBP-1a isoforms, suggesting that the HBP-1a(17) gene may be autoregulated, although the binding affinity of these proteins in vitro is very low. In Arabidopsis plants, activation of the HBP-1a(17) promoter was highly restricted to photosynthetically active mesophyll, and guard cells and vascular bundles of vegetative leaves. Etiolation of transgenic plants resulted in inhibition of expression of the HBP-1a(17) promoter. Indeed, the HBP-1a(17) promoter contains several sequence elements homologous to cis-acting elements conserved in light inducible promoters. It is, therefore, assumed that the HBP-1a(17) gene is light regulated and that HBP-1a(17) is involved in light-responsive gene transcription via the G-box. PMID- 7476858 TI - Analysis of an insertional operator mutation (gntOi) that affects the expression level of the Bacillus subtilis gnt operon, and characterization of gntOi suppressor mutations. AB - The Bacillus subtilis gnt operon is negatively regulated via interaction of the gnt repressor (GntR) with an operator upstream of gntR, which is antagonized by gluconate. An 8 bp insertional operator mutation (gntOi) of the gnt operon was constructed which affected the expression level of this operon. Two suppressors of this gntOi mutation, exhibiting normal expression, were also isolated; one involved a threonine substitution for the Ala-48 residue (gntR48T) within the helix-turn-helix DNA-binding motif of GntR, and the other an adenine substitution for the guanine at nucleotide -4 within the gntOi operator (gntOiM4A) (+ 1 is the transcription initiation site). The gntR48T mutation by itself rendered the gnt operon partially constitutive. When the gntR43L mutation, which renders the gnt operon fully constitutive, was introduced into the gntOi or gntOiM4A mutant, the operator mutations were found not to affect the promoter activity of the gnt operon. These in vivo results indicate that the gntOi mutation affects the operator interaction with GntR, causing a low expression level even in the presence of gluconate. In vitro gel retardation and DNase I footprint analyses demonstrated that even when gluconate was present, GntR still bound to the gntOi operator region. PMID- 7476859 TI - The promoter of the gene Itr1 from barley confers a different tissue specificity in transgenic tobacco. AB - Tissue-specific expression of the gene coding for trypsin inhibitor BTI-CMe in barley (Itr1) occurs during the first half of endosperm development. In transgenic tobacco, the Itr1 promoter drives expression of the beta-glucuronidase reporter gene not only in developing endosperm but also in embryo, cotyledons and the meristematic intercotyledonary zone of germinating seedlings. A promoter fragment extending 343 bp upstream of the translation initiation ATG codon was sufficient for full transgene expression, whereas, the proximal 83 bp segment of the promoter was inactive. Possible reasons for the differences in expression patterns are discussed. PMID- 7476860 TI - Translational coupling and limited degradation of a polycistronic messenger modulate differential gene expression in the parD stability system of plasmid R1. AB - The parD stability system of plasmid R1 is an auto-regulated operon containing two genes, kis and kid, that code, respectively, for a killer protein (Kid) and for an antagonist of Kid action (Kis protein). A polycistronic transcript and a shorter mRNA, coding only for Kis and ending in a stem-loop sequence, have been identified as the main parD transcripts in cells carrying a derepressed parD operon. In this communication we show that both parD mRNAs have a half-life close to 1 min and are present in similar amounts. Using an assay based on cell-free extracts of Escherichia coli, we demonstrate that the short kis mRNA originates from limited degradation of the bicistronic parD transcript and that the stem loop structure within the 5' end of the kid gene is specifically required for the formation of this short transcript. In vivo experiments show that synthesis of Kis is required for efficient synthesis of Kid. These data indicate that RNA processing and translational coupling are important mechanisms that modulate the differential expression of the two genes, kis and kid, in the bicistronic parD operon. PMID- 7476861 TI - Identification of Streptomyces violaceoruber Tu22 genes involved in the biosynthesis of granaticin. AB - A 50 kb region of DNA from Streptomyces violaceoruber Tu22, containing genes encoding proteins involved in the biosynthesis of granaticin, was isolated. The DNA sequence of a 7.3 kb fragment from this region, located approximately 10 kb from the genes that encode the polyketide synthetase responsible for formation of the benzoisochromane quinone skeleton, revealed five open reading frames (ORF1 ORF5). The deduced amino acid sequence of GraE, encoded by ORF2, shows 60.8% identity (75.2% similarity) to a dTDP-glucose dehydratase (StrE) from Streptomyces griseus. Cultures of Escherichia coli containing plasmids with ORF2, on a 2.1 kb BamHI fragment, were able to catalyze the formation of dTDP-4-keto-6 deoxy-D-glucose from dTDP-glucose at 5 times the rate of control cultures, confirming that ORF2 encodes a dTDP-glucose dehydratase. The amino acid sequence encoded by ORF3 (GraD) is 51.4% identical (69.9% similar) to that of StrD, a dTDP glucose synthase from Streptomyces griseus. The amino acid sequence encoded by ORF4 shares similarities with proteins that confer resistance to tetracycline and methylenomycin, and is suggested to be involved in transporting granaticin out of the cells by an active efflux mechanism. PMID- 7476862 TI - Functional analysis of the Drosophila CDC2 Dm gene in fission yeast. AB - The cdc2+ gene product (p34cdc2) is a protein kinase that regulates entry into mitosis in all eukaryotic cells. The role that p34cdc2 plays in the cell cycle has been extensively investigated in a number of organisms, including the fission yeast Schizosaccharomyces pombe. To study the degree of functional conservation among evolutionarily distant p34cdc2 proteins, we have constructed a S. pombe strain in which the yeast cdc2+ gene has been replaced by its Drosophila homologue CDC2Dm (the CDC2Dm strain). This CDC2Dm S. pombe strain is viable, capable of mating and producing four viable meiotic products, indicating that the fly p34CDC2Dm recognizes all the essential S. pombe cdc2+ substrates, and that it is recognized by cyclin partners and other elements required for its activity. The p34CDC2Dm protein yields a lethal phenotype in combination with the mutant B type cyclin p56cdc13-117, suggesting that this S. pombe cyclin might interact less efficiently with the Drosophila protein than with its native p34cdc2 counterpart. This CDC2Dm strain also responds to nutritional starvation and to incomplete DNA synthesis, indicating that proteins involved in these signal transduction pathways, interact properly with p34CDC2Dm (and/or that p34cdc2 independent pathways are used). The CDC2Dm gene produces a 'wee' phenotype, and it is largely insensitive to the action of the S. pombe wee1+ mitotic inhibitor, suggesting that Drosophila wee1+ homologue might not be functionally conserved. This CDC2Dm strain is hypersensitive to UV irradiation, to the same degree as wee1-deficient mutants. A strain which co-expresses the Drosophila and yeast cdc2+ genes shows a dominant wee phenotype, but displays a wild-type sensitivity to UV irradiation, suggesting that p34cdc2 triggers mitosis and influences the UV sensitivity by independent mechanisms. PMID- 7476863 TI - An Escherichia coli curved DNA-binding protein whose expression is affected by the stationary phase-specific sigma factor sigma S. AB - From Escherichia coli, a DNA-binding protein that preferentially recognizes a curved DNA sequence was isolated and shown to correspond to one that has recently been reported as a binding protein for the replication origin of the E. coli chromosome, named Rob. Here, a rob promoter-lacZ transcriptional fusion was constructed on the chromosome, and used to demonstrate that the expression of rob is notably enhanced at the onset of stationary phase in Luria-broth and also under certain growth conditions in a minimal medium, such as glucose- and phosphate-starvation medium. It was further shown that this growth condition dependent expression of rob is notably reduced in a null mutant for the stationary phase-specific sigma subunit of RNA polymerase, sigma s, although sigma s-independent expression of rob was significant during the logarithmic growth phase. Furthermore the rob null mutant was found to exhibit, as compared with the wild-type, an altered profile of protein synthesis, particularly at the very late stationary phase. PMID- 7476864 TI - Nucleotide sequence of the recF gene cluster from Staphylococcus aureus and complementation analysis in Bacillus subtilis recF mutants. PMID- 7476865 TI - A hotspot of spontaneous and UV-induced illegitimate recombination during formation of lambda bio transducing phage. AB - To study the mechanism of spontaneous and UV-induced illegitimate recombination, we examined the formation of the lambda bio specialized transducing phage in Escherichia coli. Because most lambda bio transducing phages have double defects in the red and gam genes and have the capacity to form a plaque on an E. coli P2 lysogen (Spi- phenotype), we selected lambda bio transducing phage by their Spi- phenotype, rather than using the bio marker. We determined sequences of recombination junctions of lambda bio transducing phages isolated with or without UV irradiation and deduced sequences of parental recombination sites. The recombination sites were widely distributed on E. coli bio and lambda DNAs, except for a hotspot which accounts for 57% of UV-induced lambda bio transducing phages and 77% of spontaneously induced lambda bio transducing phages. The hotspot sites on E. coli and lambda DNAs shared a short homology of 9 bp. In addition, we detected direct repeat sequences of 8 bp within and near both the bio and lambda hotspots. A recA mutation did not affect the frequency of the recombination at the hotspot, indicating that this recombination is not a variant of recA-dependent homologous recombination. We discuss a model in which the short homology as well as the direct repeats play essential roles in illegitimate recombination at the hotspot. PMID- 7476866 TI - A selection system for diploid and against haploid cells in Schizosaccharomyces pombe. AB - We have isolated a mutant of Schizosaccharomyces pombe whose growth is temperature sensitive when it is haploid but not when it is diploid. This mutant may provide a useful system for selecting nonconditional mutants which are defective in diploid formation upon conjugation. PMID- 7476867 TI - Transgene inactivation in Petunia hybrida is influenced by the properties of the foreign gene. AB - Petunia mutant RL01 was transformed with maize A1 and gerbera gdfr cDNAs, which both encode dihydroflavonol-4-reductase (DFR) activity. The same Agrobacterium vector and the same version of the CaMV 35S promoter were used in both experiments. Transformation with the cDNAs resulted in production of pelargonidin pigments in the transformants. However, the A1 and gdfr transformants showed clearly different phenotypes. The flowers of the primary A1 transformants were pale and showed variability in pigmentation during their growth, while the flowers of the gdfr transformants showed intense and highly stable coloration. The color difference in the primary transformants was reflected in the expression levels of the transgenes as well as in the levels of anthocyanin pigment. As previously reported by others, the instability in pigmentation in the A1 transformants was more often detected in clones with multiple copies of the transgene and was associated with methylation of the 35S promoter and of the transgene cDNA itself. In the gdfr transformants, the most intense pigmentation was observed in plants with multiple transgenes in their genome. Only rarely was partial methylation of the 35S promoter detected, while the gdfr cDNA always remained in an unmethylated state. We conclude that the properties of the transgene itself strongly influence the inactivation process. The dicotyledonous gdfr cDNA with a lower GC content and fewer possible methylation sites is more 'compatible' the genomic organization of petunia and this prevents it being recognized as a foreign gene and hence silenced by methylation. PMID- 7476869 TI - The promoter from the rice nuclear gene encoding chloroplast aldolase confers mesophyll-specific and light-regulated expression in transgenic tobacco. AB - The rice genome contains at least four separate loci that encode aldolase isozymes. Among these, the aldolase P (AldP) gene, a nuclear gene coding for chloroplast aldolase, is expressed predominantly in the leaf blade mesophyll cells in rice. To dissect promoter elements that regulate such tissue- or cell type-specific expression, we constructed various AldP promoter-beta-glucuronidase (GUS) fusion genes and transferred them into Nicotiana tabacum (tobacco) plants. Analysis of GUS activities in the transgenic tobacco revealed the presence of at least two elements within 2.0 kb AldP promoter region. One is located within the segment from position -2.0 kb to -1.2 kb and acts as a negative element. The other is a positive element located between -1.2 kb and -0.31 kb that confers developmentally regulated, mesophyll cell-specific expression. In addition, the 1.2 kb rice promoter segment flanking the transcription start site contains an element(s) that serves as target for light induction in tobacco. The results suggest that the AldP gene promoter of rice, a monocot promoter, can function in an essentially physiological manner in the dicot tobacco plant. PMID- 7476868 TI - Arabidopsis thaliana tryptophan synthase alpha: gene cloning, expression, and subunit interaction. AB - The tryptophan synthase alpha subunit catalyzes the conversion of indole-3 glycerolphosphate to indole, the penultimate reaction in the biosynthesis of the essential amino acid tryptophan. A cDNA encoding Arabidopsis thaliana tryptophan synthase alpha(TSA1) was isolated by complementation of an Escherichia coli delta trpA mutation and by polymerase chain reaction amplification from a cDNA library using degenerate primers. A TSA1 genomic clone was also isolated and 5 kb of the DNA sequence determined. A single sequence in the Arabidopsis genome with homology to the TSA1 cDNA was detected by high-stringency genomic Southern blot hybridization. In contrast under hybridization conditions of reduced stringency, one or two additional homologous sequences were observed. A 1.4 kb transcript was detected in wild-type RNA with the TSA1 cDNA as a probe. Several lines of evidence, including immunoaffinity chromatography, suggest that the active A. thaliana tryptophan synthase enzyme consists of a heterosubunit complex, presumably analogous to the prokaryotic alpha 2 beta 2 complex. Immunoblot analysis indicated that the plant alpha and beta subunits are present throughout development. PMID- 7476870 TI - Assembly of nucleosomal DNA in a cell-free extract from wild-type and top1- strains of Ustilago maydis. AB - An in vitro nucleosome assembly system has been established from cell-free extracts of the fungus Ustilago maydis. The extract catalyzed DNA supercoiling in the absence of exogenously added co-factors such as ATP and MgCl2 and was inhibited by moderate concentrations (200 mM) of KCl or NaCl. DNA supercoiling occurs via the formation of nucleosomes. Similar extracts, displaying the same activity, were prepared from Saccharomyces cerevisiae and Candida albicans, suggesting that the extract preparation protocol may be useful for many lower eukaryotic systems. An extract prepared from a strain of U. maydis lacking topoisomerase I failed to catalyze nucleosome assembly, clearly implicating this enzyme in this process. Addition of purified topoisomerase I, and, to a lesser extent, topoisomerase II, to the top1- extract regenerated the supercoiling activity. Our results provide a method for preparing assembly extracts from organisms, that are particularly amenable to genetic manipulation. PMID- 7476871 TI - MMK2, a novel alfalfa MAP kinase, specifically complements the yeast MPK1 function. AB - Mitogen-activated protein (MAP) kinases are serine/threonine protein kinases that are activated in response to a variety of stimuli. Here we report the isolation of an alfalfa cDNA encoding a functional MAP kinase, termed MMK2. The predicted amino acid sequence of MMK2 shares 65% identity with a previously identified alfalfa MAP kinase, termed MMK1. Both alfalfa cDNA clones encode functional kinases when expressed in bacteria, undergoing autophosphorylation and activation to phosphorylate myelin basic protein in vitro. However, only MMK2 was able to phosphorylate a 39 kDa protein from the detergent-resistant cytoskeleton of carrot cells. The distinctiveness of MMK2 was further shown by complementation analysis of three different MAP kinase-dependent yeast pathways; this revealed a highly specific replacement of the yeast MPK1(SLT2) kinase by MMK2, which was found to be dependent on activation by the upstream regulators of the pathway. These results establish the existence of MAP kinases with different characteristics in higher plants, suggesting the possibility that they could mediate different cellular responses. PMID- 7476872 TI - Expression of the dnaB gene of Escherichia coli is inducible by replication blocking DNA damage in a recA-independent manner. AB - The replicative DNA helicase encoded by the dnaB gene is essential for chromosomal DNA replication in Escherichia coli. The DnaB protein is a component of the phi X-type primosome which is regarded as a model system for lagging strand synthesis of the chromosome. Using translational lacZ fusions at the plasmid and chromosomal levels, we studied the influence of DNA-damaging agents on dnaB gene expression. We found that DNA damage caused by mitomycin C, methyl methanesulphonate, 4-nitro-quinoline N-oxide, and UV irradiation led to a moderate, but significant induction of dnaB gene expression. Comparative S1 analysis of transcripts in untreated and induced cells demonstrated that the induction is due to increased transcription from the dnaB promoter. In contrast to other DNA damage-inducible replication genes, such as dnaA, dnaN, dnaQ, and polA, expression of which is not inducible in recA and lexA mutants, the induction of dnaB was also observed in a recA1 mutant. These results show that the induction of dnaB gene expression by replication-blocking DNA damage is due to a mechanism other than the indirectly SOS-dependent induction of the other DNA replication genes. Moreover, the data suggest that replication proteins are involved in recovery from replication-blocking DNA damage in two different ways- on the one hand at the level of initiation and on the other hand at the level of elongation. PMID- 7476873 TI - The tef1 box, a ubiquitous cis-acting element involved in the activation of plant genes that are highly expressed in cycling cells. AB - In Arabidopsis thaliana, the tef1 box is a cis-acting promoter element of the EF 1 alpha A1 gene involved in the activation of transcription in meristematic tissues. The initiation of root calli in transgenic Arabidopsis by 2,4-D shows that the tef1-dependent expression of the GUS reporter gene is not restricted to meristematic regions but involves all of the cycling cells. Hybridization experiments conducted using Arabidopsis cDNA clones organized in a dense array on filters, and cDNA probes prepared from cells in various states of growth, or blocked at different steps of the cell cycle, indicate that the enhanced expression of EF-1 alpha genes occurs in cycling cells at the point of entry into the cell cycle and remains constant during transit through the cycle. The analysis of several promoters of genes, other than EF-1 alpha, which are overexpressed in growing cells and involved in the processes of translation or redox regulation, reveals the presence of sequences showing partial homologies with the tef1 box. The Arabidopsis ribosomal gene srp18 and the tobacco gene thioh2, encoding a thioredoxin h, contain such sequences. Gel retardation experiments suggest that these sequences are targets for the same proteins as those that interact with the tef1 box of the Arabidopsis EF-1 alpha A1 gene. In transfected Arabidopsis protoplasts, the putative tef1 sequence thioh2 partially restores the activity of a tef1 box-less EF-1 alpha A1 promoter. These data demonstrate that the tef1 box is a ubiquitous cis-acting element involved in the transcriptional activation of plant genes that are overexpressed in cycling cells.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7476874 TI - Dosage suppressors of the dominant G1 cyclin mutant CLN3-2: identification of a yeast gene encoding a putative RNA/ssDNA binding protein. AB - Three G1 cyclins, CLN1, CLN2, and CLN3, have been identified in the budding yeast Saccharomyces cerevisiae. G1 cyclins are essential, albeit functionally redundant, rate-limiting activators of cell cycle initiation. We have isolated dosage-dependent suppressor genes (designated HMD genes) of the mating defect caused by CLN3-2, a dominant mutation in CLN3, HMD2 and HMD3 are identical to STE4 and STE5, respectively, HMD1 is an essential gene that encodes a protein containing a putative RNA binding domain. Overproduction of HMD1 results in a relatively specific reduction in the level of the CLN3 or CLN3-2 transcript. This reduction occurs subsequent to transcription initiation of CLN3 since overexpression of HMD1 did not affect expression of a heterologous transcript from the CLN3 promoter but did result in a reduction of CLN3 transcript expressed from a heterologous promoter. HMD1 has at least one essential role independent of its effect on CLN3 since HMD1 remains essential for viability in the absence of a functional CLN3 gene. PMID- 7476875 TI - Characterization of the let-653 gene in Caenorhabditis elegans. AB - A mutation in the let-653 gene of Caenorhabditis elegans results in larval death. The lethal arrest is concurrent with the appearance of a vacuole anterior to the lower pharyngeal bulb. The position of the vacuole is consistent with a dysfunction of the secretory/excretory apparatus. Germline transformation rescue experiments were able to position the let-653 gene to two overlapping cosmid subclones. Sequence data generated from both cDNA and genomic DNA subclones indicated that let-653 encodes a mucin-like protein. Our characterization suggests that a mucin-like protein is essential for effective functioning of the secretory/excretory apparatus within C. elegans. PMID- 7476876 TI - Heat shock and light activation of a Chlamydomonas HSP70 gene are mediated by independent regulatory pathways. AB - Induction of HSP70 heat shock genes by light has been demonstrated in Chlamydomonas. Our aim was to establish whether this induction by light is mediated by the heat stress sensing pathway or by an independent signal chain. Inhibitors of cytoplasmic protein synthesis revealed an initial difference. Cycloheximide and other inhibitors of protein synthesis prevented HSP70A induction upon illumination but not during heat stress. Analysis of HSP70A induction in cells that had differentiated into gametes revealed a second difference. While heat shock resulted in elevated HSP70A mRNA levels, light was no longer able to serve as an inducer in gametes. To identify the regulatory sequences that mediate the response of the HSP70A gene to either heat stress or light we introduced a series of progressive 5' truncations into its promoter sequence. Analyses of the levels of mRNA transcribed from these deletion constructs showed that in most of them the responses to heat shock and light were similar, suggesting that light induction is mediated by a light-activated heat shock factor. However, we show that the HSP70A promoter also contains cis-acting sequences involved in light induction that do not participate in induction by heat stress. Together, these results provide evidence for a regulation of HSP70A gene expression by light through a heat shock-independent signal pathway. PMID- 7476877 TI - Multiple outer membrane receptors for uptake of ferric pseudobactins in Pseudomonas putida WCS358. AB - Under iron limitation Pseudomonas putida WCS358 produces a fluorescent siderophore, pseudobactin 358, which, after complexing iron, is transported back into the cell via the specific outer membrane receptor PupA. In addition, this strain has the capacity to take up iron via a large variety of siderophores produced by other fluorescent pseudomonads. Putative receptor genes for such siderophores were identified in the chromosome of strain WCS358 by PCR using primers matching two domains conserved in four ferric pseudobactin receptors, including PupA. Eleven amplification products within the expected size range were obtained. Sequence analysis confirmed that the products were derived from genes encoding outer membrane receptors. Two complete receptor genes were isolated from a genomic library of P. putida WCS358. Both protein products are involved in the transport of a limited number of specific ferric pseudobactins. These results indicate that the ability of P. putida WCS358 to exploit many different heterologous pseudobactins is related to the presence of multiple outer membrane receptor proteins. PMID- 7476878 TI - Comparative mapping in grasses. Wheat relationships. AB - Conventionally, the genetics of species of the family Gramineae have been studied separately. Comparative mapping using DNA markers offers a method of combining the research efforts in each species. In this study, we developed consensus maps for members of the Triticeae tribe (Triticum aestivum, T. tauschii, and Hordeum spp.) and compared them to rice, maize and oat. The aneuploid stocks available in wheat are invaluable for comparative mapping because almost every DNA fragment can be allocated to a chromosome arm, thus preventing erroneous conclusions about probes that could not be mapped due to a lack of polymorphism between mapping parents. The orders of the markers detected by probes mapped in rice, maize and oat were conserved for 93, 92 and 94% of the length of Triticeae consensus maps, respectively. The chromosome segments duplicated within the maize genome by ancient polyploidization events were identified by homoeology of segments from two maize chromosomes to regions of one Triticeae chromosome. Homoeologous segments conserved across Triticeae species, rice, maize, and oat can be identified for each Triticeae chromosome. Putative orthologous loci for several simply inherited and quantitatively inherited traits in Gramineae species were identified. PMID- 7476880 TI - Localization of P2X purinoceptor transcripts in the rat nervous system. AB - We used transcript-specific oligonucleotides to examine the localization in the rat nervous system of the corresponding mRNAs for the two P2X purinoceptor genes cloned recently from the rat vas deferens and PC12 cells. PC12 P2X purinoceptor mRNA was labeled in the olfactory tubercle, striatum, hypothalamus, hippocampus, dentate gyrus, amygdala, cortex, and cerebellum, whereas the vas deferens P2X purinoceptor-specific probes labeled the cerebellum and, at lower levels of expression, the striatum, hippocampus, and cortex. Both types of P2X purinoceptor transcript were found on cell bodies in the nodose and superior cervical ganglia. The presence of these two purinoceptor transcripts in the brain was confirmed by polymerase chain reaction. Two partial cDNAs, identical to sections of the PC12 or vas deferens P2X purinoceptor coding sequences, were amplified from neonatal brain and cerebellum poly(A)+ RNA, respectively. These findings are in broad agreement with earlier Northern blot studies on the PC12 P2X purinoceptor mRNA but differ from those for the vas deferens P2X purinoceptor mRNA, which had not previously been detected in adult brain. This difference is attributed to the low levels seen in the adult compared with the neonate and to the greater sensitivity of the methods used in the present study. The neonate medial habenula had low levels of transcripts for the PC12 but none for the vas deferens P2X purinoceptor. Because pharmacologically the recombinant PC12 P2X purinoceptor differs from the functional purinoceptor in the medial habenula, these results suggest the existence of other, unidentified, P2X purinoceptors in the rat nervous system. PMID- 7476879 TI - The raspberry locus encodes Drosophila inosine monophosphate dehydrogenase. AB - Investigation of an enhancer-trap line exhibiting testis-specific beta galactosidase expression led to the isolation of the Drosophila gene encoding inosine monophosphate dehydrogenase (IMPD), the rate-limiting enzyme in guanine nucleotide synthesis, which has been implicated in cell cycle control and malignant transformation. Northern and in situ hybridization analysis demonstrated that the gene has a complex expression pattern involving several independently regulated transcripts. Two ubiquitous, but highly ovary enriched, transcripts of 2.5 and 1.9 kb are expressed in the nurse cells and delivered to the oocyte, whilst a 0.9 kb transcript is found exclusively in the testis. The 2.5 kb transcript encodes a 58 kDa protein, which is highly similar in length and sequence to mouse and human IMPDs and is presumably required for GTP synthesis during early embryogenesis. Over-expression of this cDNA in Escherichia coli yielded a product of the predicted size, which was demonstrated to possess IMPD activity in a spectrophotometric assay. The coding capacity of the other transcripts is currently uncertain. We present evidence that IMPD is the product of the raspberry (ras) locus at 9E and the functions of the gene are discussed in relation to the phenotypes of ras mutants. PMID- 7476881 TI - A discrete site for general anesthetics on a postsynaptic receptor. AB - General anesthetics depress central nervous system excitability via a mechanism that probably involves effects on synaptic ion channels, but the fundamental molecular nature of the site where they act is unknown. Although the importance of hydrophobicity for general anesthetic drug potency has long been established, it remains uncertain whether these "nonspecific" drugs act on membrane proteins directly or by modification of the physical properties of the lipid membrane or the lipid-protein interface. We find that specific mutations in the acetylcholine receptor pore-forming M2 domains enhance the sensitivity of the receptor to the general anesthetics isoflurane, hexanol, and octanol, suggesting that these agents act by binding directly to a discrete protein site at or near these residues. The sensitivity of the receptor to block by general anesthetics increases with increased hydrophobicity of these residues, demonstrating that hydrophobic forces dominate the interaction of drugs with their protein site. Furthermore, octanol inhibits both wild-type and mutant nicotinic acetylcholine receptors preferentially after channel opening, which is consistent with a mechanism where drugs bind within the receptor's pore. Similar sites on postsynaptic ion channels in brain may represent general anesthetic targets for modulating consciousness. PMID- 7476882 TI - Involvement of a tyrosine kinase pathway in the growth-promoting effects of angiotensin II on aortic smooth muscle cells. AB - Angiotensin II (AII) is a growth factor that stimulates protein synthesis and induces cellular hypertrophy in aortic smooth muscle cells (SMC). This trophic effect is mediated by the AT1 subtype of AII receptors. However, very little is known about the cellular signaling pathways involved in this response. In the present study, we examined the role of protein tyrosine phosphorylation in the growth-promoting effects of AII on rat aortic SMC. The addition of AII to quiescent aortic SMC induced tyrosine phosphorylation of multiple substrates, as revealed by antiphosphotyrosine immunoblotting. This response was blocked by preincubation with the AT1-selective antagonist losartan. To explore the functional role of this signaling pathway, we performed experiments with two mechanistically distinct tyrosine kinase inhibitors. Treatment of quiescent aortic SMC with genistein and herbimycin A abolished the stimulatory effect of AII on overall protein tyrosine phosphorylation. Similarly, the two inhibitors prevented AII-induced tyrosine phosphorylation of the cytoskeletal protein paxillin. Under the same conditions, incubation with genistein or herbimycin A did not interfere with AII binding to the AT1 receptor and did not significantly affect AII-stimulated inositol-1,4,5-trisphosphate production and Ca2+ mobilization. In parallel to their selective action on tyrosine phosphorylation, both genistein and herbimycin A completely inhibited AII-stimulated protein synthesis in a dose-dependent manner. In contrast, the two inhibitors were much less potent in preventing the trophic effect of phorbol-12-myristate 13-acetate in these cells. We further demonstrate that genistein and herbimycin A did not prevent mitogen-activated protein kinase activation and c-fos gene induction, which is consistent with the notion that these downstream effectors do not link AII-induced tyrosine phosphorylation to protein synthesis. These results provide evidence that tyrosine phosphorylation has a critical role in cellular hypertrophy and is involved in AII action in vascular SMC. PMID- 7476883 TI - Phosphorylation of cyclic AMP response element-binding protein and induction of c fos gene expression on withdrawal from chronic treatment with carbachol in NG108 15 cells. AB - Alterations in adenylyl cyclase activity in cultured cells after prolonged exposure to drugs such as morphine have been extensively studied as models for drug tolerance and withdrawal. NG108-15 cells develop increased intracellular cAMP concentrations after abrupt withdrawal from chronic treatment with the muscarinic cholinergic agonist carbachol. To determine whether this withdrawal induced increase in cAMP modifies gene expression, we studied phosphorylation of the cAMP response element-binding protein (CREB) and expression of the c-fos gene, known to contain a cAMP response element, in NG108-15 cells after abrupt withdrawal from chronic treatment with carbachol. Prostaglandin E1, which activates adenylyl cyclase, caused concentration-dependent increases in the phosphorylation of CREB and in the abundance of c-fos mRNA. These changes occurred with small increments in cAMP accumulation. In cells treated with carbachol for 48 hr, induction of withdrawal with the muscarinic antagonist atropine led to a small increase in intracellular cAMP concentration but an 11.6 fold increase in the phosphorylation of CREB and a 3.4-fold increase in accumulation of c-fos mRNA. The adenylyl cyclase inhibitor 2',5' dideoxyadenosine, which attenuated the chronic carbachol-induced increase in cAMP concentration, prevented the increased phosphorylation of CREB and the enhanced accumulation of c-fos mRNA during atropine-induced withdrawal. These results indicate that expression of the c-fos gene is induced by the small increments in cAMP concentration that can occur in cells on withdrawal from chronic treatment with drugs such as muscarinic agonists. PMID- 7476884 TI - Angiotensin II down-regulates the vascular smooth muscle AT1 receptor by transcriptional and post-transcriptional mechanisms: evidence for homologous and heterologous regulation. AB - The vascular angiotensin II (ANG II) receptor (AT1) is a central component of the renin-angiotensin system; thus, regulation of its expression is likely to be important in cardiovascular responsiveness. We demonstrate that ANG II down regulates its receptor in rat aortic vascular smooth muscle cells. Incubation for 4 hr with 100 nM ANG II decreased AT1 mRNA and protein by 70% and 35%, respectively. This homologous down-regulation was concentration and time dependent and was blocked by the AT1 antagonist losartan. It did not appear to be mediated by protein kinase C or other protein kinases but was dependent on the sustained signaling pathway sensitive to phenylarsine oxide. Heterologous down regulation was observed with the agonists alpha-thrombin and ATP and the cAMP increasing agent forskolin. ANG II inhibited transcription by 50% and destabilized the AT1 mRNA. Down-regulation of AT1 mRNA was blocked by transcription and translation inhibitors, suggesting that it required expression of a protein factor or factors. These results indicate that ANG II down-regulates its vascular receptor by both transcriptional and post-transcriptional mechanisms. Homologous and heterologous down-regulation of the AT1 receptor may participate in the coordinated physiological adaptation of vascular tone to vasoactive hormones. PMID- 7476885 TI - Differential effects of a topoisomerase I inhibitor on dioxin inducibility and high-level expression of the cytochrome P450IA1 gene. AB - The basic helix-loop-helix containing dioxin receptor mediates dioxin signal transduction. The ligand-activated receptor complex binds to specific sequences termed xenobiotic response elements and regulates transcription of target genes such as the gene for cytochrome P450IA1. This study demonstrates that induction of cytochrome P450IA1 and P450IB1 gene expression by a dioxin receptor ligand is repressed by camptothecin, an inhibitor of the topoisomerase I enzyme. However, a transiently transfected reporter construct under control of an xenobiotic response element-containing promoter was not affected by the topoisomerase inhibitor. In agreement with this observation, ligand-dependent activation of the dioxin receptor to its DNA-binding form is not altered by camptothecin as analyzed by electrophoretic mobility shift assay. Moreover, the inhibitory effect of camptothecin cannot be exerted once the P450IA1 gene has been activated. These results imply that topoisomerase I activity is necessary for the primary P450IA1 induction response, possibly involving dioxin-dependent alterations in chromatin structure of the P450IA1 promoter. PMID- 7476886 TI - Activation and selective inhibition of a cyclic AMP-specific phosphodiesterase, PDE-4D3. AB - Prostaglandin E2 produces a transient increase in the intracellular concentration of cAMP in a human promonocytic cell line (U937). The temporal pattern consists of a rapid increase followed by a gradual decline to a new steady state. The decline phase coincides with an increase in the activity of a high affinity form of cAMP phosphodiesterase (PDE). Immunoprecipitation with specific antibodies revealed that the activated enzyme is a variant of PDE-4D. To confirm this observation, three isoforms of human PDE-4 (A, B, and D) were cloned and expressed in Sf9 cells with recombinant baculovirus infection. The activity of only one of the isoforms (PDE-4D3) increased after incubation with the catalytic subunit of protein kinase A and Mg-ATP. Hydrolytic activity of human PDE-4D3 was dependent on Mg2+. Before phosphorylation, the concentration-response curve for Mg2+ was biphasic and ranged from 0.1 to 100 mM. Phosphorylation of PDE-4D3 by protein kinase A produced a monophasic Mg2+ response curve (0.5 Vmax = 0.2 mM). Phosphorylation of PDE-4D3 increased the sensitivity of the enzyme to inhibition by RS-25344 (approximately 100-fold) and RS-33793 (approximately 330-fold). Thus, phosphorylation of PDE-4D3 induces an apparent conformation change that increases maximum velocity and sensitivity to inhibition by some analogues of nitraquazone. These observations provide the basis for a novel pharmacological strategy that targets an activated form of PDE in human leukocytes. Selective PDE-4D3 inhibitors may have useful anti-inflammatory properties with fewer adverse side effects than other PDE-4 inhibitors. PMID- 7476887 TI - Presynaptic Ca2+/calmodulin-dependent protein kinase II: autophosphorylation and activity increase in the hippocampus after long-term blockade of serotonin reuptake. AB - It is known that long-term treatment with antidepressants induces an enhancement of neurotransmission in the pathway projecting from raphe nuclei to the hippocampus. In the case of selective serotonin (5-HT) reuptake inhibitors, this enhancement is due to a desensitization of presynaptic 5-HT autoreceptors and a concomitant increase in 5-HT release in terminal areas. To investigate whether this effect is accompanied by adaptive changes in the molecular machinery regulating transmitter release at serotonergic terminals, autophosphorylation and activity of Ca2+/calmodulin-dependent protein kinase II were measured in subsynaptosomal fractions from hippocampus and total cortex. Long-term treatment with two selective serotonin reuptake inhibitors (paroxetine and fluvoxamine) and with a nonselective reuptake inhibitor (venlafaxine) induces a large increase of kinase autophosphorylation in synaptic vesicles and synaptic cytosol in the hippocampus but not in synaptosomal membranes. No significant change was detected in total cortex. The change is not reproduced by the direct addition of the drugs to the phosphorylation system and is not elicited by acute treatment of the animals. The increase in autophosphorylation is not accounted for by neosynthesis or translocation of the kinase to synaptic terminals. The change is restricted to the kinase located inside the terminals and is not detected in synaptosomal membranes, containing predominantly postsynaptic kinase, suggesting that only presynaptic kinase is affected. In the same fractions, the kinase activity is increased. These results are in agreement with reports suggesting a presynaptic effect for the SSRIs and disclose a new putative site of action for psychotropic drugs. PMID- 7476888 TI - Developmental and endocrine regulation of P450 isoforms in rat breast. AB - Cytochrome P450 was partially purified from rat breast tissue from 1-, 2-, 3-, 6 , 9-, and 15-week-old pregnant, lactating, or 3-week postlactation rats. The detergent-solubilized P450 was spectrally quantified, and the P450 isozyme pattern in the different samples was characterized by Western blot analysis with antibodies against cytochromes P450 1A1, 1A2, 2A, 2B, 2D4, 3A, 4A, 2E, and 19. The yield of P450 was 5-60 pmol/g wet weight tissue, with the highest yields in 1 and 2-week-old pups and lactating rats. The cytochromes P450 expressed in the breast can be divided into six main groups on the basis of their pattern of regulation: (a) those present in all samples (4A, 2E1, and 2D4), (b) those highly expressed in 1- to 3-week-old rats (2D4 and 3A), (c) those expressed only after 9 weeks of age [P450 19 (aromatase)], (d) those induced in pregnancy and maintained during lactation (1A1), (e) those induced in pregnancy and not maintained during lactation (2A), and (f) those induced 3 weeks after lactation (2B, 2A, and 3A). Reverse transcription-polymerase reaction amplification was used to confirm the presence of P450 isoforms in the breast. The mRNAs of cytochromes P450 1A1, 2A1, 2B1-3, 2D1, 2D3, 2D4, 2E1, and 4A3 were detected on analysis of total breast RNA. The mRNA of CYP 3A1 was not convincingly detected in untreated rat breast but was inducible by treatment with pregnenolone-16-alpha-carbonitrile. The presence of these various forms of P450 in the breast and their regulation by age and endocrine status may have implications for in situ metabolism of steroids and steroid antagonists and for activation of procarcinogens. PMID- 7476889 TI - Modulation of detoxification gene expression in human colon HT29 cells by glutathione-S-transferase inhibitors. AB - We investigated the effects of glutathione-S-transferase (GST) inhibitor treatment on human colon HT29 cell mRNA levels of dihydrodiol dehydrogenase (DDH), glyoxalase I, and gamma-glutamylcysteine synthetase. Time- and concentration-dependent increases in both DDH and gamma-glutamylcysteine synthetase mRNAs resulted from treatment with ethacrynic acid, ethacrynic acid/glutathione conjugate, and T.199 (gamma-glutamyl-S-(benzyl)-cysteinyl-R(-) phenyl glycine diethyl ester), a selective GST pi inhibitor. In contrast, glutathione analogue GST alpha- and GST mu-selective inhibitors did not induce expression of these genes. Treatment with ethacrynic acid or T.199 had no effect on the mRNA levels of the glutathione-dependent glyoxalase I gene. Pretreatment of cells with buthionine-DL-sulfoximine, a gamma-glutamylcysteine synthetase inhibitor and glutathione depleter, coupled with ethacrynic acid, ethacrynic acid/glutathione conjugate, or T.199 resulted in greater levels of gamma glutamylcysteine synthetase and DDH induction compared with single treatments. Treatment with buthionine-DL-sulfoximine alone resulted in modest increases in both gamma-glutamylcysteine synthetase and DDH expression. Analyses of DDH induction by both differential Northern hybridization with specific oligonucleotides as probes and reverse transcriptase-polymerase chain reaction amplification of products, followed by diagnostic restriction digestion with endonucleases, showed that ethacrynic acid induced multiple DDH transcripts in HT29 cells and human HepG2 and SKHep1 hepatoma cells. Possible induction mechanisms include the alteration of sulfhydryl status by the electrophilic properties of EA or by elevations of endogenously generated oxidative stress via transient removal of GST pi from the cytosolic GST pool. PMID- 7476890 TI - Cloning and expression of a human metabotropic glutamate receptor 1 alpha: enhanced coupling on co-transfection with a glutamate transporter. AB - We cloned and expressed a human metabotropic glutamate receptor 1 alpha (HmGluR1 alpha) in a novel cell line. The human mGluR1 alpha cDNA was found to be 86% identical to rat mGluR1 alpha, and the predicted protein sequence was found to be 93% identical to rat mGluR1 alpha. We expressed HmGluR1 alpha in AV12-664, an adenovirus-transformed Syrian hamster cell line. To prevent tonic activation of HmGluR1 alpha by glutamate that may be released by these cells into the extracellular medium, HmGluR1 alpha was co-expressed in AV12-664 cells with a rat glutamate/aspartate transporter (GLAST). This allowed investigation of the effect that clearance of glutamate from the extracellular space would have on HmGluR1 alpha function. A comparison of mRNA levels revealed that HmGluR1 alpha was similarly expressed in cells with or without co-expression of GLAST. However, HmGluR1 alpha-mediated phosphoinositide hydrolysis was efficiently elicited only in cells co-expressing rat GLAST. Blockade of glutamate transport by L-trans pyrrolidine-2,4-dicarboxylic acid resulted in an increase in glutamate levels in the media and an increase in basal HmGluR1 alpha-mediated phosphoinositide hydrolysis. Long-term pretreatment of cells with L-trans-pyrrolidine-2,4 dicarboxylic acid resulted in media glutamate levels similar to those in cells not expressing GLAST. However, this resulted in a dramatic decrease in 1 aminocyclopentane-1S,3R-dicarboxylic acid- and glutamate-stimulated phosphoinositide hydrolysis. These studies suggest that co-expression of mGluR1 alpha with a glutamate transporter prevents desensitization of the receptor, thus achieving optimal coupling of the receptor with its effector system. PMID- 7476891 TI - Eukaryotic DNA topoisomerases mediated DNA cleavage induced by a new inhibitor: NSC 665517. AB - A compound with a novel structure, NSC 665517, was tested in the National Cancer Institute Preclinical Drug Discovery Screen. With the COMPARE algorithm, the pattern of differential cytotoxicity for NSC 665517 most closely resembled those of known topoisomerase II (top2) inhibitors. In vitro data showed that NSC 665517 induced DNA cleavage in the presence of top2 and topoisomerase I (top1) (at a higher concentration). The minimum concentration required to induce top2 cleavage was 0.5 microM. A substantial decrease in top2-induced cleavage by NSC 665517 was seen when the reaction mixtures were shifted to elevated temperature (55 degrees), suggesting that top2-induced cleavage occurs through the mechanism of stabilizing the reversible enzyme/DNA complex and inhibiting religation. The DNA cleavage pattern induced by NSC 665517 with top2 was different than that of other known top2 inhibitors, including etoposide, mitoxantrone, anthracyclines, amsacrine, and ellipticine. top2 cleavage sites induced by NSC 665517 showed strong preference for G located 3' to the top2-mediated DNA cleavage (position +1). NSC 665517 produced limited DNA unwinding at high drug concentration. DNA damage analyzed in KB cells by alkaline elution showed that NSC 665517 induced strand break. Data from the cytotoxicity in KB-V1 overexpressing P-glycoprotein and COMPARE analysis with rhodamine efflux assay indicated that NSC 665517 is a substrate of P-glycoprotein. These results strongly suggest that NSC 665517 is a novel topoisomerase-targeted drug. Preclinical evaluation of NSC 665517 as an antitumor agent is under way. PMID- 7476893 TI - Yeast cysteine proteinase gene ycp1 induces resistance to bleomycin in mammalian cells. AB - Tumor resistance to the glycopeptide anticancer drug bleomycin (BLM) has been suggested to involve metabolic inactivation by BLM hydrolase. Direct evidence for this hypothesis is lacking due to difficulties in obtaining full-length BLM hydrolase cDNA from mammalian cells. In the present investigation, we used the yeast cysteine proteinase gene ycp1, a homologue of the mammalian BLM hydrolase gene, to provide direct evidence of the importance of BLM metabolism in BLM resistance. Transfection of ycp1 into NIH 3T3 cells induced resistance of these cells to BLM. The ycp1-transfected cells also metabolized BLM A2 to its inactive metabolite deamido-BLM A2 to a much greater extent. The ycp1-induced BLM resistance was completely reversed by the cysteine proteinase inhibitor E-64, a known inhibitor of BLM hydrolase. Transfection of NIH 3T3 cells with the plasmid pUT533-Sh ble, a bacterial BLM resistance gene that encodes a 14-kDa protein that does not metabolize BLM, also induced BLM resistance, but this resistance was not overcome by E-64. The results demonstrate that increased BLM hydrolase activity in NIH 3T3 cells causes BLM resistance and that inhibition of BLM metabolism sensitizes these cells to BLM. Thus, the molecular approach described in the present study directly implicates BLM hydrolase in BLM resistance. PMID- 7476892 TI - Pharmacological subtypes of the gamma-aminobutyric acidA receptors defined by a gamma-aminobutyric acid analogue 4,5,6,7-tetrahydroisoxazolo[5,4-c] pyridin-3-ol and allosteric coupling: characterization using subunit-specific antibodies. AB - Various alpha and beta 3 subunit-specific antibodies were used to characterize some of the heterogeneous ligand-binding properties of gamma-aminobutyric acidA receptors. Polyclonal antibodies that were raised against the cytoplasmic amino acid sequence (380-392) of the rat beta 3 subunit recognized a single polypeptide of molecular mass of 58 kDa in Western blots with Ro7-1986 affinity-purified GABAA receptors from the rat brain, and a doublet of molecular mass of 54 kDa and 56 kDa in receptors from the bovine cortex, hippocampus, and cerebellum. Deglycosylation of purified receptors from the bovine cortex with N-glycanase resulted in a single band immunostained at molecular mass of 52 kDa. These anti beta 3 subunit antibodies immunoprecipitated approximately 50% of [3H]flunitrazepam binding sites from soluble extracts of bovine cortex, whereas beta cyto antibodies, which probably recognize all beta subunit isoforms, precipitated almost 100% of benzodiazepine binding sites. These results indicate heterogeneity of GABAA receptor subunit composition with respect to the nature of beta subunits. The GABA analogue 4,5,6,7-tetrahydroisoxazolo[5,4-c]pyridin-3-ol (THIP), like GABA, shows heterogeneous binding affinities in brain homogenates. The higher affinity sites were previously suggested as corresponding to a 58-kDa polypeptide in rat that is photoaffinity-labeled with [3H]muscimol, a band that comigrates with the one stained by anti-beta 3 antibodies. However, THIP affinity was not significantly different between receptors containing beta 3 subunits and those lacking beta 3, as demonstrated by similar affinities in receptors that ere immunoprecipitated by anti-beta 3 antibodies and those that were not. Also, THIP displaced [3H]muscimol binding with similar multiple affinities across brain regions where different beta subunit variants are expressed with varying abundances. These observations suggest that the property of high affinity THIP binding cannot be explained solely by beta 3 subunits. The coupling efficiency between GABA and benzodiazepine binding sites appears to be determined by the nature of alpha subunits rather than of beta subunits. GABA enhanced [3H]flunitrazepam binding with different efficacies and potencies in receptors immunoprecipitated by anti-alpha 1, -alpha 2, and -alpha 3 subunit antibodies. In contrast, beta 3 subunit-enriched and disenriched receptors did not differ in this property. [3H]Flunitrazepam binding in GABAA receptors containing alpha 2 and alpha 3 subunits was enhanced to a significantly greater extent than were those with alpha 1. In addition, receptors containing alpha 1 and alpha 3 subunits had higher potencies of enhancement than did those with alpha 2 subunits.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 7476894 TI - A structure-function relationship among reserpine and yohimbine analogues in their ability to increase expression of mdr1 and P-glycoprotein in a human colon carcinoma cell line. AB - We previously showed that there is a structure-function relationship among reserpine and yohimbine analogues in their ability to inhibit the function of P glycoprotein (P-gp) and reverse multidrug resistance (MDR). Because some P-gp inhibitors (e.g., verapamil and nifedipine) can increase mdr1 and P-gp expression in human colon carcinoma cell lines, we used our reserpine/yohimbine analogues to determine whether there was a structural requirement for this induction. We found that 10 microM reserpine increased both mdr1 and P-gp expression by 4-10-fold in 48 hr in a human colon carcinoma cell line that expresses moderate levels of mdr1 (LS180-Ad50) but not in several other cell lines that expressed no mdr1. The reserpine/yohimbine analogues rescinnamine, trimethoxybenzoylyohimbine, and LY191401 (compound G), all of which contain the three structural elements used to describe the MDR pharmacophore, also increased both mdr1 and P-gp expression significantly. Despite some exceptions, we found that there was a good association between the ability of these analogues to induce mdr1 and P-gp expression and their ability to reverse vinblastine and doxorubicin resistance, revealing a structure-function relationship for this phenomenon. The increased P gp expressed by these cells appeared to be functional, as determined by flow cytometric detection of rhodamine 123 retention. The increased expression was suppressed by 5,6-dichloro-1-beta-D-ribofuranosylbenzimidazole, an RNA synthesis inhibitor, whereas the protein synthesis inhibitor cycloheximide enhanced the expression several-fold, suggesting that induction of mdr1 by these analogues is regulated at both the transcriptional and post-transcriptional levels.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7476895 TI - Reversible modulation of opioid receptor binding in intact neural cells by endogenous guanosine triphosphate. AB - Incubation of SH-SY5Y neural cells with mycophenolic acid (MPA), an inhibitor of inosine monophosphate dehydrogenase (the key enzyme in purine nucleotide biosynthesis), reduced the cellular content of GTP by 94% relative to its concentration in control cells (43 nmol/mg protein) without altering the level of GDP. Although in GTP-depleted intact cells the receptor binding parameters (Kd and Bmax) of the opioid antagonist [3H]naltrexone were unchanged from those in untreated cells, the binding affinity of the mu-selective opioid agonist [3H]Tyr D-Ala-Gly-(Me)- Phe-Gly-ol ([3H]DAMGO) was enhanced 2-fold. Furthermore, the kinetics of ligand/receptor interaction revealed that in the nucleotide-depleted cells, the dissociation rate constant for [3H]DAMGO was reduced by 44%. Initial exposure of SH-SY5Y cells to pertussis toxin reduced high-affinity ligand binding by 95% and abolished the effect of MPA treatment. Renewed incubation of the GTP depleted cells with guanosine restored the original GTP levels and agonist binding. Neither MPA nor guanosine treatment changed the Bmax of [3H]DAMGO binding. Forskolin- and prostaglandin E1-stimulated adenylyl cyclase activities were decreased significantly in GTP-depleted cells. DAMGO and [D-Pen2,D Pen5]enkephalin inhibitions of adenylyl cyclase were also affected with MPA treatment. Maximal inhibition of forskolin-stimulated adenylyl cyclase activity by both of the agonists was reduced, whereas MPA caused a 2-fold reduction in potency for DAMGO. The results show that reduction in endogenous GTP levels leads to noticeable changes in agonist, receptor, and G protein interactions, as measured by agonist binding, and to subsequent diminution of the signal transduction, as reflected by the cAMP levels. PMID- 7476896 TI - Competition between positive and negative allosteric effectors on muscarinic receptors. AB - Alcuronium allosterically increases the affinity of cardiac muscarinic receptors for methyl-N-scopolamine (NMS), whereas gallamine has the opposite effect. We discovered that strychnine also increases the affinity of muscarinic receptors in rat heart atria for NMS. It is not known whether the positive and the negative allosteric effectors bind to the same binding site. To investigate this question, we elaborated on a theoretical model predicting changes in the binding of a classic radiolabeled ligand occurring in the presence of a positive and a negative allosteric effector that compete for the allosteric binding site. The model is based on data obtained at equilibrium and avoids uncertainties associated with the use of nonequilibrium methods for the evaluation of interactions between allosteric ligands. We examined changes in the binding of [3H]NMS to membranes of rat heart atria exposed to various concentrations of a positive allosteric effector (alcuronium or strychnine) and of a negative allosteric effector (gallamine) simultaneously. The binding data obtained were in perfect agreement with the model assuming competition between gallamine and alcuronium and gallamine and strychnine, strongly suggesting that these positive and negative allosteric effectors bind to identical or overlapping sites. PMID- 7476897 TI - Structural and ligand recognition properties of imidazoline binding proteins in tissues of rat and rabbit. AB - Imidazoline/guanidinium receptive sites (IGRS) belong to a family of membrane proteins that selectively recognize certain pharmacologically active compounds with an imidazoline or a guanidinium moiety. The role of such proteins in the cellular responses elicited by these compounds is unclear, but two members of this protein family are identical to isoforms of monoamine oxidase, an enzyme involved in the metabolism of monamine neurotransmitters. To characterize the structural and ligand recognition properties of the imidazoline binding proteins, we used the photoaffinity adduct [125I]iodoazidophe-noxymethylimidazoline ([125I]AZIPI) to label their ligand binding subunits in selected target tissues (kidney, pancreatic B cells, liver, and salivary gland). Photoaffinity labeling of membrane preparations or subcellular particulate fractions from various rat, rabbit, or hamster tissues indicated two labeled peptides of M(r) approximately 55,000 and approximately 61,000, the relative tissue distribution of which mirrored the expression of the A or B isoforms of monoamine oxidase. The ligand binding subunit of imidazoline binding proteins was identified on two peptides of M(r) approximately 55,000 and approximately 61,000 in rat and rabbit kidney, rat liver, rabbit salivary gland, and the pancreatic B cell line RIN-5AH, whereas only an M(r) approximately 61,000 peptide was observed in rat salivary gland and the hamster pancreatic B cell line HIT-T15. Saturation labeling experiments indicated that [125I]AZIPI exhibited similar affinity (Kd approximately 2-3 nM) for both the M(r) approximately 55,000 and approximately 61,000 peptides. However, competitive inhibition of photolabeling indicated that the two peptides were distinguished by their affinity for the guanidinium guanabenz or their interaction with potassium. Although some types of imidazoline binding sites are located on the enzyme monoamine oxidase, the nonisoform selective enzyme inhibitor pargyline did not alter photoaffinity labeling of either the M(r) approximately 55,000 or approximately 61,000 peptide, indicating that imidazolines/guanidiniums and active site inhibitors of monoamine oxidase interact with different domains on the enzyme. In rat kidney and liver, an additional photolabeled peptide of M(r) approximately 25,000 was observed, and its ligand recognition profile was distinct from the M(r) approximately 55,000 and approximately 61,000 species. In contrast with the mitochondrial location of the larger peptides, subcellular fractionation of liver homogenates indicated that the M(r) approximately 25,000 localized to the plasma membrane. PMID- 7476898 TI - Two classes of structurally different antagonists display similar species preference for the human tachykinin neurokinin3 receptor. AB - Two classes of structurally different tachykinin neurokinin3 (NK3) antagonists were used to evaluate species difference in antagonist binding between human and rat NK3 receptors. In competition binding experiments with [125I-MePhe7]NKB as radioligand, PD 154740, PD 157672, SR 48968, and SR 142801 displayed lower Ki values for the human NK3 receptor (40 +/- 4, 12 +/- 1,350 +/- 50, and 0.40 +/- 0.05 nM, respectively) than for the rat NK3 receptor (2450 +/- 130, 288 +/- 25, > 10,000, and 11.0 +/- 0.5 nM, respectively). Data from in vitro functional assay showed similar species preference as observed with the competition binding assay. It was shown previously that substitution of only two amino acid residues in the rat receptor to their human counterparts could change the species selectivity of SR 48968, a weak NK3 antagonist. In the double-substituted rat mutant, all three antagonists (PD 154740, PD 157672, and SR 142801) displayed Ki values (76 +/- 8, 16 +/- 2, and 0.50 +/- 0.05 nM, respectively) very similar to the Ki values for the wild-type human NK3 receptor. Thus, in addition to their previously reported effects on SR 48968, these two amino acid residues are responsible for the species selectivity of these three additional NK3 antagonists. Because PD 154740 and PD 157672 are very different structurally from SR 48968 and SR 142801, our results indicate that the two identified residues may be involved in adopting a receptor conformation that favors the binding of NK3 antagonists that display species preference for the human NK3 receptor. PMID- 7476899 TI - Glycine modulates ethanol inhibition of heteromeric N-methyl-D-aspartate receptors expressed in Xenopus oocytes. AB - Ethanol inhibits N-methyl-D-aspartate (NMDA) receptor-mediated responses at pharmacologically relevant concentrations, suggesting that inhibition of NMDA receptors may underlie some of the actions of ethanol in the central nervous system. We examined the ability of glycine to modulate ethanol inhibition of four recombinant heteromeric NMDA receptors (NR1a/NR2A through NR2D) expressed in Xenopus oocytes. Ethanol dose-response analysis revealed enhanced inhibitory efficacy of ethanol in the presence of subsaturating glycine concentrations at the NR1/NR2A, NR1/NR2C, and NR1/NR2D receptors. When assayed over a range of glycine concentrations, ethanol exhibited both glycine-reversible and glycine independent inhibition of NMDA receptors. In contrast, ethanol inhibition of recombinant NMDA receptors was independent of NMDA concentration. Glycine reversal of ethanol inhibition suggested that ethanol might lower the affinity of glycine for the NMDA receptor and thereby decrease response magnitude. Consistent with this hypothesis, ethanol significantly reduced glycine affinity at NR1/NR2A and NR1/NR2C receptors. Evaluation of the glycine-independent component of ethanol inhibition demonstrated that in the presence of saturating concentrations of glycine, the NR1/NR2A and NR1/NR2B receptors were more sensitive to ethanol than the NR1/NR2C and NR1/NR2D receptors. Activation of the NR1/NR2D heteromers by NMDA and low concentrations of glycine elicited responses characterized by an initial peak followed by a lower-amplitude plateau response, which is consistent with glycine-sensitive desensitization as previously described for native NMDA receptors. In addition, nondesensitizing NR1/NR2B responses elicited in the presence of subsaturating concentrations of glycine were frequently converted into desensitizing responses by the addition of ethanol, an effect that was reversed with increasing glycine concentrations. The ability of ethanol to promote glycine-sensitive desensitization further suggests an interaction between glycine and ethanol inhibition of the NMDA receptor. Taken together, the results of the present report demonstrate that ethanol inhibition of NMDA receptors has both glycine-reversible and glycine-independent components, suggesting two distinct molecular mechanisms for ethanol inhibition of NMDA receptors. PMID- 7476900 TI - Structural requirements of analogues of polyamines for migration and growth of IEC-6 cells. AB - Healing of gastrointestinal mucosal lesions occurs through two processes: an early one involving cell migration and a later one in which cell division replaces lost cells. Both processes require the presence of polyamines, but the mechanism of action of these compounds is unknown. In the present study, we examined the ability of analogues of spermidine and spermine to support migration and growth of IEC-6 cells that have been grown in alpha-difluoromethylornithine to inhibit polyamines. All analogues of spermidine with the general formula x-3 (referring to the numbers of carbon atoms on either side of the central nitrogen), where x = 2-12, competed with spermidine for entry into the cells. However, in addition to spermidine (x = 4), only compounds for which x = 2, 3, or 6 supported migration and only those for which x = 2 or 7 supported growth. Spermine analogues 3-x-3, for which x = 3, 6, 9, or 12, competed for entry into the cells, but only compounds for which x = 3 or 6 supported migration and only the compound for which x = 3, in addition to spermine (x = 4), supported growth. In addition, analogues 2-3-2, 3-2-3, and 2-(3)2, a branched compound, supported both migration and growth but entered the cell via a mechanism different than that for spermidine and spermine. These data define some of the specific structural requirements for polyamines to produce their physiological effects. PMID- 7476901 TI - D2 dopamine receptor antisense oligodeoxynucleotide inhibits the synthesis of a functional pool of D2 dopamine receptors. AB - In vivo administration of an antisense oligonucleotide targeted toward the D2 dopamine (DA) receptor mRNA (D2 AS) markedly inhibited D2 receptor-mediated behaviors but produced only a relatively small reduction in the levels of D2 DA receptors in mouse striatum. This apparent dissociation between DA receptor mediated behaviors and the levels of D2 DA receptors was addressed by inhibiting the total number of D2 DA receptors by intraperitoneal administration of the selective, irreversibly acting D2 DA receptor antagonist fluphenazine-N-mustard (FNM) and then determining the effects of D2 AS, administered intracerebroventricularly, on the rate of synthesis of D2 DA receptors and on the recovery of D2 receptor-mediated behaviors. FNM inactivated approximately 90% of D2 DA receptors within 4 hr of treatment, after which the receptors returned to normal levels by approximately 8 days. D2 AS treatment significantly inhibited the rate of recovery of D2 DA receptors in striatum of FNM-treated mice. FNM treatment also produced a number of behavioral alterations, including catalepsy, and the inhibition of stereotypic behavior induced by the D2/D3 DA receptor agonist quinpirole. Both of these behaviors returned to normal within 8 days after FNM treatment. D2 AS treatment delayed the restoration of these FNM-induced behaviors. Thus, it reduced the rate of disappearance of the cataleptic behavior induced by FNM and significantly delayed the restoration of the stereotypic behavior induced by quinpirole. The changes induced by D2 AS on D2 receptor mediated behaviors were reversed on cessation of D2 AS treatment. A random oligomer given in the same amount and for the same length of time as that of the D2 AS had no significant effects on either D2 DA receptor synthesis or DA receptor-mediated behaviors. These studies demonstrate that in vivo administration of D2 AS decreased the rate of recovery of D2 DA receptors and inhibited the recovery of D2 DA receptor-mediated behaviors after irreversible receptor inactivation and suggest that D2 AS treatment inhibits the synthesis of a functional pool of D2 DA receptors. PMID- 7476902 TI - Contribution of sequence and phosphorothioate content to inhibition of cell growth and adhesion caused by c-myc antisense oligomers. AB - c-myc is overexpressed in glioblastoma multiforme, the most common form of brain tumor. To find a suitable target for in vivo antisense therapy of gliomas, we investigated the biological effects on the human glioma cell line, U87MG, of antisense oligonucleotides targeted against the translation start site of c-myc mRNA. Parameters examined included c-myc protein level, cell proliferation, and cell adhesion to substratum. Oligonucleotides were administered by electroporation as capped phosphorothioates. Antisense oligomers caused a reduction in c-myc protein expression, loss of cell adhesion to plastic, and complete growth inhibition. Various control sequences, including sense, scrambled, and three-base mismatched oligomers, were also tested. Some of the controls retained a dG quartet found in the antisense sequence. Reduction in c myc protein and cell growth and loss of cell adhesion were specific to the antisense sequence. Surprisingly, fully thioated antisense and scrambled sequences, either containing or lacking a dG quartet, were equally inhibitory to both cell growth and adhesion. Loss of cell adhesion was observed with only phosphorothioate-containing oligomers, not with either their phosphodiester or nuclease-resistant PA congeners, and was completely reversed when cells were plated onto fibronectin. These results demonstrate that a commonly used c-myc antisense oligomer also displays dramatic, sequence- but not antisense-specific effects on cell proliferation and cellular adhesion, depending on the backbone. PMID- 7476903 TI - Suppression of lipopolysaccharide-induced tumor necrosis factor-alpha generation from human peripheral blood monocytes by inhibitors of phosphodiesterase 4: interaction with stimulants of adenylyl cyclase. AB - We assessed the role of cyclic nucleotides in modulating lipopolysaccharide (LPS) induced tumor necrosis factor-alpha (TNF-alpha) generation in human peripheral blood monocytes. Exposure of monocytes to LPS (3 ng/ml) evoked a delayed, time dependent generation of TNF-alpha that reached a maximum level 5-6 hr after LPS challenge and remained constant for up to 24 hr. This effect was concentration dependent and resulted in a 20-40-fold increase in the release of TNF-alpha that was sensitive to actinomycin D and cycloheximide. Treatment of monocytes with agents reputed to activate the cAMP/cAMP-dependent protein kinase (PKA) cascade in general inhibited LPS-induced TNF-alpha generation. Thus, the beta 2 adrenoceptor agonists albuterol and procaterol partially (approximately 40%) suppressed TNF-alpha generation in a propranolol-sensitive manner. Furthermore, 8 bromo-cAMP, cholera toxin, prostaglandin E2, and a number of drugs (i.e., rolipram (ZK 62711), denbufylline (BRL 30892), Ro 20-1724, benafentrine (AH 21 132), that inhibit the phosphodiesterase (PDE) 4 isoenzyme family abolished cytokine generation. In contrast, forskolin, inhibitors of PDE3 and PDE5, and activators of soluble and particulate guanylyl cyclase were essentially inactive. Interestingly, rolipram failed to potentiate the inhibitory effect of albuterol on LPS-induced TNF-alpha biosynthesis but, paradoxically, synergized with albuterol in the generation of cAMP and in the activation of PKA. When PGE2 was used to activate adenylyl cyclase, however, rolipram potentiated cAMP accumulation, PKA activation, and inhibition of TNF-alpha generation. In contrast, forskolin did not increase the cAMP content of monocytes in the absence or presence of rolipram. Collectively, these data suggest that LPS-induced TNF alpha generation by human peripheral blood monocytes is due to increased transcription and subsequent translation of the TNF-alpha gene and that these effects are suppressed by a range of agents that activate the cAMP/PKA cascade. However, the failure of rolipram to potentiate the inhibitory effect of albuterol and procaterol on TNF-alpha generation suggests that beta 2-adrenoceptor agonists may affect gene expression and/or post-transcriptional regulatory processes by, at least in part, a cAMP-independent mechanism(s). PMID- 7476904 TI - Dihydrofolate reductase binding and cellular uptake of nonpolyglutamatable antifolates: correlates of cytotoxicity toward methotrexate-sensitive and resistant human head and neck squamous carcinoma cells. AB - Several mechanisms have been demonstrated to be independently involved in methotrexate (MTX) resistance, including increased dihydrofolate reductase (DHFR) activity, decreased membrane transport, and decreased conversion to noneffluxing polyglutamates by folylpolyglutamate synthetase. We conducted the present study to test the hypothesis that nonpolyglutamatable antifolates with an N delta hemiphthaloyl-L-ornithine side chain could be more potent than MTX against MTX sensitive and -resistant human carcinoma cells via tighter DHFR binding, more efficient cellular uptake, the ability to bypass defective polyglutamation, or a combination. Two nonpolyglutamatable antifolates, N alpha-(4-amino-4 deoxypteroyl)-N delta-hemiphthaloyl-L-ornithine (PT523) and the new B-ring analogue N alpha-[4-[N-(2,4-diamino-5-chloroquinazolin-6-yl)methyl]amino] benzoyl N delta-hemiphthaloyl-L-ornithine (PT619), were tested as inhibitors of purified recombinant human DHFR and were found to bind somewhat better to the enzyme than MTX as determined by competitive radioligand binding assay. PT523 and PT619 were 9- and 14-fold, respectively, more active than MTX as inhibitors of parental SCC25 human and neck squamous carcinoma cell growth in 72-hr cultures. Moreover, there was an even greater increase in relative potency against two previously described MTX-resistant cell lines with an increased DHFR content and a decreased ability to convert MTX to polyglutamates: SCC25/R1 (selected with MTX) and SCC25/CP (selected with cisplatin but collaterally resistant to MTX). Both PT523 and PT619 very efficiently inhibited [3H]MTX uptake by SCC25 cells in a 1-hr assay, with PT523 being 11-fold more potent and PT619 being 17-fold more potent than MTX. Greater inhibition of [3H]MTX uptake with PT523 and PT619 than with MTX was also observed in SCC25/R1 and SCC/CP cells. However, the increase in activity of PT523 and PT619 relative to MTX in uptake experiments was less than that in growth-inhibition assays, especially for SCC25/CP cells. This suggested that additional cytotoxicity determinants may exist in these resistant cells. PMID- 7476905 TI - Lysophosphatidic acid regulation of cyclic AMP accumulation in cultured human airway smooth muscle cells. AB - The effects of the simple bioactive lipid mediator lysophosphatidic acid (LPA) on cAMP accumulation were investigated in cultured human airway smooth muscle cells (ASMC). Pretreatment of cells with LPA induced an increase in subsequent stimulation of cAMP accumulation by forskolin and by isoproterenol. When included during the assay of cAMP accumulation rather than as a pretreatment, LPA inhibited forskolin stimulation but enhanced isoproterenol stimulation. Both effects of LPA on forskolin stimulation were completely blocked by pertussis toxin treatment, whereas the effects on isoproterenol stimulation appeared relatively insensitive to pertussis toxin. The protein kinase C activator phorbol 12-myristate-13-acetate (PMA) sensitized forskolin stimulation to a similar extent as did LPA, and the combination of LPA plus PMA caused markedly more sensitization than either agent alone. In contrast, PMA inhibited isoproterenol stimulation and markedly decreased the sensitization induced by LPA. Serum also induced sensitization, and sensitization by LPA plus serum was no greater than that with LPA alone. LPA-induced sensitization appeared to be independent of protein kinase C activation because it was unchanged in cells treated to down regulate protein kinase C. LPA also stimulated polyphosphoinositide hydrolysis, and this stimulation was partially inhibited by pertussis toxin treatment. These results suggest that LPA activates receptors coupled to both the pertussis toxin sensitive G protein Gi and the pertussis toxin-insensitive G protein Gq. The complex effects of LPA, PMA, and pertussis toxin on cAMP accumulation in these cells are consistent with the expression of the type 2 isozyme of adenylyl cyclase in these cells. PMID- 7476906 TI - Comparative pharmacology of epibatidine: a potent agonist for neuronal nicotinic acetylcholine receptors. AB - Pharmacological properties of the (+)- and (-)-isomers of synthetic epibatidine, exo-2-(6-chloro-3-pyridyl)-7-azabicyclo-[2.2.1]heptane, were compared with nicotine and acetylcholine on several subtypes of chicken and human nicotinic acetylcholine receptors (AChRs). Both isomers of epibatidine behaved as extremely potent full agonists on chicken (alpha 3 beta 2, alpha 3 beta 4, alpha 4 beta 2, alpha 7, and alpha 8) and human (alpha 3 beta 2, alpha 3 beta 4, and alpha 7) neuronal AChRs expressed in Xenopus oocytes. Currents induced by epibatidine were effectively blocked by the nicotinic antagonists hexamethonium and mecamylamine. Apparent affinity was 100 to 1000-fold higher for epibatidine than for nicotine or acetylcholine. EC50 values ranged from 1 nM (for homomeric chicken alpha 8) to 2 microM (for homomeric chicken alpha 7). Epibatidine showed comparatively lower affinity for muscle-type AChRs from Torpedo and humans (EC50 values, 1.6 and 16 microM respectively). In binding assays, epibatidine was used on AChR subtypes immunoisolated from chicken brain and retina (alpha 4 beta 2, alpha 7, and alpha 8), the human neuronal cell line SH-SY5Y (alpha 3 and alpha 7), Torpedo electric organ (alpha 1 beta 1 gamma delta), or the human rhabdomyosarcoma cell line TE671 (alpha 1 beta 1 gamma delta). Both isomers of epibatidine exhibited extremely high affinity for all neuronal AChRs tested, with KI values ranging from 0.6 pM (human alpha 3 AChRs) to 0.6 microM (chicken alpha 7 AChRs). In contrast, epibatidine had lower affinity for Torpedo muscle-type AChRs (KI approximately 5 microM). Racemic [3H]epibatidine was an effective labeling reagent for human alpha 3 beta 2 AChRs, exhibiting a KD (0.14 nM) similar to the KI values observed for unlabeled (+)-epibatidine (0.23 nM) or (-)-epibatidine (0.16 nM). PMID- 7476907 TI - Mutation of Asp100 in the second transmembrane domain of the cholecystokinin B receptor increases antagonist binding and reduces signal transduction. AB - We examined the functional significance of two residues present in the second (Asp100) and seventh (Asn391) transmembrane domains of the rat cholecystokininB (CCKB) receptor that are highly conserved among the members of the G protein coupled receptor family. Substitution of Asn for Asp100 by using site-directed mutagenesis did not change the affinity and selectivity for agonists but slightly increased the affinity of three CCKB-selective antagonists of different chemical structures. Cells expressing the mutant receptor exhibited a 50% reduction in CCKB-induced phosphoinositide turnover compared with cells expressing the wild type receptor, suggesting a critical role for this residue in the coupling of the CCKB receptor to G protein. This latter was shown to be insensitive to pertussis toxin treatment and could therefore belong to the Gq family. Replacement of Asn391 by Asp located in the seventh transmembrane domain did not change agonist binding or phosphoinositide turnover. This suggests that in contrast to the gonadotropin-releasing hormone receptor, there is no direct interaction in the CCKB receptor between Asp100 and Asn391. However, a rhodopsin-based molecular modeling of the CCKB receptor showed a spatial proximity between Asp100 and the carboxyl terminal part of the third intracellular loop, known to interact with G protein. This could explain the reduction in phosphoinositide turnover observed with the Asn100 mutant. PMID- 7476908 TI - Growth hormone regulation of male-specific rat liver P450s 2A2 and 3A2: induction by intermittent growth hormone pulses in male but not female rats rendered growth hormone deficient by neonatal monosodium glutamate. AB - Growth hormone (GH) secretory patterns regulate the expression of several sex dependent liver cytochrome P450 (CYP) genes. Studies using the hypophysectomized rat model have established that the intermittent plasma GH secretory pattern associated with adult male rats markedly stimulates liver expression of the male specific CYP 2C11, a testosterone 2 alpha- and 16 alpha-hydroxylase, but is not required for expression of other male-specific liver enzymes, including CYP 2A2, a testosterone 15 alpha-hydroxylase, and CYP 3A2, a testosterone 6 beta hydroxylase. In the present study, the effects of intermittent GH treatment on liver CYP expression were studied in adult rats rendered GH deficient by neonatal administration of monosodium glutamate (MSG), which depletes circulating adult GH without the global loss of other pituitary-dependent hormones that is associated with hypophysectomy. Restoration of the normal masculine circulating GH profile of six daily pulses (180-225 ng GH/ml/peak) in MSG-treated male rats by the use of an external pumping apparatus led to a substantial (30-50%) restoration of normal male levels of CYP 2A2 and CYP 3A2 activity, protein, and mRNA. GH pulsation at the nonphysiological frequencies of two or four times per day was less effective unless given at a dose that resulted in supraphysiological plasma GH levels. Although intermittent GH treatment can induce male-specific P450 expression in hypophysectomized female rats, the same hormone treatment did not stimulate CYP 2A2 or CYP 3A2 expression in MSG-treated female rats. Liver GH receptor mRNA levels at adulthood were not significantly altered by neonatal MSG treatment, suggesting that the unresponsiveness of MSG-treated females and the previously reported low responsiveness of MSG-treated males to GH-induced CYP 2C11 expression are not due to the absence of GH receptor. Moreover, normal liver IGF-1 mRNA levels were expressed in the MSG-treated female rats, suggesting that the liver GH receptor is functional in these animals. The present findings establish that the adult male-specific enzymes CYP 2A2 and CYP 3A2 can be positively regulated by intermittent GH pulsation despite their GH-independent expression in hypophysectomized rats. Moreover, neonatal MSG treatment, particularly in female rats, may lead to the loss of factors other than GH that are required for full expression of the pulsatile GH-stimulated CYP 2A2, 3A2, and 2C11 genes. PMID- 7476909 TI - Hypophosphorylation of topoisomerase II in etoposide (VP-16)-resistant human leukemia K562 cells associated with reduced levels of beta II protein kinase C. AB - We selected and characterized a 30-fold etoposide (VP-16)-resistant subline of K562 human leukemia cells (K/VP.5) that exhibits quantitative and qualitative changes in topoisomerase II, including hypophosphorylation of this drug target. The initial rate of topoisomerase II phosphorylation was reduced 3-fold in K/VP.5 compared with K562 cells, but the rate of dephosphorylation was similar. Analysis of potential topoisomerase II protein kinases revealed a 3-fold reduction in the level of the beta II protein kinase C (PKC) in K/VP.5 cells, whereas levels of alpha- and epsilon PKC, casein kinase II, p42map kinase, and p34cdc2 kinase were comparable for both cell lines. The PKC activator, bryostatin 1, together with K562 nuclear extracts potentiated VP-16-induced topoisomerase II/DNA covalent complex formation in nuclei isolated from K/VP.5 cells but not from K562 cells. Bryostatin 1 effects were blocked by the PKC inhibitor 7-O-methyl-hydroxy staurosporine. Bryostatin 1 also up-regulated topoisomerase II phosphorylation and potentiated VP-16 activity in intact K/VP.5 cells but had no enhancing effect in K562 cells. 4 beta-Phorbol-12,13-dibutyrate and 12-O-tetradecanoylphorbol-13 acetate did not potentiate VP-16-induced topoisomerase II/DNA complex formation in intact cells or in isolated K/VP.5 nuclei. Together, our results indicate that beta II PKC plays a role in modulating the VP-16-induced DNA binding activity of topoisomerase II in resistant K/VP.5 cells through a mechanism linked to phosphorylation of topoisomerase II. PMID- 7476910 TI - Species differences in A1 adenosine receptor/G protein coupling: identification of a membrane protein that stabilizes the association of the receptor/G protein complex. AB - Reconstitution experiments with purified components reproduce the basic characteristics of receptor/G protein coupling, i.e., GTP-sensitive high affinity agonist binding and receptor-promoted GTP binding. However, the interaction of agonists with the A1 adenosine receptor in rat and bovine but not human brain membranes deviates from the ternary complex model since the agonist/receptor/G protein complex cannot be dissociated by high concentrations (> or = 100 microM) of the hydrolysis-resistant analogue GTP gamma S. The reason for this phenomenon referred to as a "tight coupling mode" has remained enigmatic. We show that it is attributable to a distinct membrane protein, which we labeled the coupling cofactor. Extraction of the protein from rat brain membranes with the detergent 3[3-(cholamidopropyl)diamethylammonio]-1-propanamium increased the potency of GTP gamma S by 1000-fold. After extraction, the potency was comparable to that in human brain membrane. Detergent extracts from rat brain membranes were used to resolve the component from solubilized receptors and G protein alpha and beta gamma subunits by sequential DEAE-Sephacel chromatography and Superose gel filtration (molecular weight of approximately 150 kDa in 3[3 (cholamidopropyl)diamethylammonio]-1-propanamium). Coupling cofactor restored guanine nucleotide refractoriness in a concentration-dependent manner to both detergent-extracted rat brain membranes and, albeit with lower affinity, human brain membranes. However, in human brain extracts, cofactor activity was detectable on reconstitution with rat acceptor membranes, indicating an intrinsic difference between rat and human receptors in their ability to interact with the cofactor. With high amounts of coupling cofactor present, GTP gamma S no longer decreased but rather increased agonist affinity. Readdition of partially purified coupling cofactor to acceptor membranes reduced the rate of A1 adenosine receptor mediated G protein turnover. These observations show that the component identified traps the ternary agonist/receptor/G protein complex in a stable conformation, impedes signaling of the A1 adenosine receptor, and thereby regulates the level of signal amplification. PMID- 7476911 TI - Agonist-stimulated phosphorylation of the carboxyl-terminal tail of the secretin receptor. AB - The secretin receptor belongs to a recently recognized family of G protein coupled receptors that lack the sequence motifs typical of the beta-adrenergic receptor family. Because our understanding of the regulatory mechanisms for these receptors is largely based on the latter group, we have begun to explore these mechanisms in the secretin receptor. In the present study, we focused on receptor phosphorylation, a key mechanism of receptor desensitization. Secretin receptor phosphorylation was demonstrated in intact transiently transfected COS cells and a stable receptor-bearing Chinese hamster ovary cell line in response to stimulation with native agonist. Secretin phosphoreceptor migrated on a sodium dodecyl sulfate-polyacrylamide gel at M(r) 57,000-62,000 in its native state and at M(r) 42,000 after deglycosylation, similar to the receptor that had been affinity-labeled with 125I-[Tyr10,p-NO2-Phe22]-secretin-27. Phosphorylation occurred rapidly in a secretagogue concentration-dependent manner, with 0.1 microM secretin eliciting a 7.2-fold increase in phosphorylation after 2 min. One dimensional phosphopeptide mapping after cyanogen bromide cleavage revealed a single band of M(r) 9400, corresponding in size to the carboxyl-terminal tail domain. This identification was confirmed with a truncation mutant in which potential sites of phosphorylation in the tail were eliminated and no agonist stimulated phosphorylation was observed. Phosphoamino acid analysis of the secretin phosphoreceptor demonstrated predominance of phosphothreonine over phosphoserine (3.2:1), with no phosphotyrosine observed. Three distinct carboxyl terminal truncation mutants were constructed to each eliminate a subset of potential phosphorylation sites, and differential levels of phosphorylation were observed. Appropriate biosynthetic processing, expression on the cell surface, and signaling for each of these constructs were ensured by demonstration of ligand binding and cAMP responsiveness. Thus, receptors in the recently described secretin receptor family are phosphorylated in response to agonist stimulation in a manner analogous to the beta-adrenergic receptor, likely representing an important molecular mechanism for receptor desensitization. PMID- 7476912 TI - Role of oxidative stress generated from the mitochondrial electron transport chain and mitochondrial glutathione status in loss of mitochondrial function and activation of transcription factor nuclear factor-kappa B: studies with isolated mitochondria and rat hepatocytes. AB - Mitochondria are an important source of reactive oxygen intermediates because they are the major consumers of molecular oxygen in cells. Respiration is associated with toxicity, which is related to the activation of oxygen to reactive intermediates. The purpose of the present study was to examine the role of reduced glutathione (GSH) in the maintenance of mitochondrial functions during oxidative stress induced through selective inhibition of the complex III segment of the electron transport chain. Hydrogen peroxide monitored by the fluorescence of dichlorofluorescein increased in a time- and dose-dependent manner on incubation of mitochondria with antimycin A (AA), an inhibitor of complex III. However, blockade of complex I or II with rotenone or thenoyltrifluoroacetone, respectively, did not result in accumulation of hydrogen peroxide. Depletion of mitochondrial GSH to 10-20% of control by preincubation with diethylmaleate (0.8 mM) or ethacrynic acid (250 microM) also increased dichlorofluorescein and malondialdehyde levels and resulted in an additional (2-3-fold) increase after AA. Similar results were obtained when mitochondrial GSH depletion was produced by treatment with buthionine L-sulfoximine before mirochondria isolation. The endogenous oxidative stress induced by AA was accompanied by a moderate loss of activity of ATPase complex (77% of control) and complex IV of respiration (75% of control), which was accentuated after depletion of mitochondrial GSH (51% and 45% of control, respectively). Similar results were observed in isolated hepatocytes in which depletion of mitochondrial GSH and AA led to peroxidation and mitochondrial dysfunction. In addition, with electrophoretic mobility shift assay of the transcription factor nuclear factor-kappa B (NF-kappa B), we detected its activation in response to AA (2-3-fold). Depletion of mitochondrial GSH in hepatocytes (20% of control) led to further enhancement of NF-kappa B activation (2-4-fold), which correlated with generation of hydrogen peroxide. Thus, our results suggest that GSH protects mitochondria against the endogenous oxidative stress produced at the ubiquinone site of the electron transport chain. Mitochondrial GSH depletion potentiates oxidant-induced loss of mitochondrial functions. Oxidant stress in mitochondria can promote extramitochondrial activation of NF-kappa B and therefore may affect nuclear gene expression. PMID- 7476913 TI - Subunit composition determines picrotoxin and bicuculline sensitivity of Drosophila gamma-aminobutyric acid receptors. AB - Few gamma-aminobutyric acid (GABA) receptor subunits have been cloned from insects. These include Resistance to dieldrin, or Rdl, and a homologue of the vertebrate GABAA receptor beta subunit. Unlike most vertebrate GABAA receptor subunits, Rdl forms a highly functional homomultimeric receptor. This receptor is picrotoxin (PTX) sensitive but bicuculline (BIC) insensitive and cannot be readily classified within the known GABAA receptor subtypes. In contrast, functional expression of the beta subunit homologue has not been reported. We report that coinfection of cells with recombinant baculoviruses containing Rdl plus beta subunits induces GABA receptors with distinct pharmacological and kinetic properties. Coinfection produces two separate receptor populations: one highly sensitive to PTX but BIC insensitive (Rdl homomultimers) and the other PTX insensitive and BIC sensitive (Rdl plus beta heteromultimers). Putative Rdl plus beta channels also show reduced GABA sensitivity, slow desensitization, rapid bursting, and shorter mean open time. These studies not only localize PTX and BIC sensitivity to two distinct GABA receptor subunits but also demonstrate assembly of two highly divergent GABA receptor subunits. Furthermore, the difference in channel conductance and gating between in vivo and recombinant channels implies the existence of uncharacterized GABA receptor subunits in Drosophila. PMID- 7476914 TI - Pharmacological properties of recombinant human N-methyl-D-aspartate receptors comprising NR1a/NR2A and NR1a/NR2B subunit assemblies expressed in permanently transfected mouse fibroblast cells. AB - The pharmacological properties of two recombinant human N-methyl-D-aspartate (NMDA) receptor subtypes, comprising either NR1a/NR2A or NR1a/NR2B subunits permanently transfected into mouse L(tk-) cells, have been compared using whole cell voltage-clamp electrophysiology. Glutamate was a full agonist at both receptors, having a modestly but statistically significant (p < 0.002) higher affinity for the NR2B- than the NR2A-containing receptor (microscopic Kd [mKd] = 0.76 and 0.43 microM, respectively). In comparison to glutamate, NMDA, quinolinic acid, and cis-2,3-piperidinedicarboxylic acid were partial agonists at both receptor subtypes. Maximal amplitude currents resulted when glutamate-site agonists were combined with either glycine or D-serine; both of these amino acids were, therefore, defined as full agonists at the glycine site. Glycine had an approximately 10-fold higher affinity (p < 0.0001) for NR2B- than for NR2A containing receptors (mKd = 0.057 and 0.53 microM, respectively). D-Cycloserine, (+)-(3R)-3-amino-1-hydroxypyrrolidin-2-one, (+)-cis-(4R)-methyl-(3R)-amino-1 hydroxypyrrolidin-2-one, and 1-aminocyclobutanecarboxylic acid also had higher affinities for the NR2B-containing receptor but were partial agonists, at both receptor subtypes, unlike glycine. Agonist-evoked whole-cell currents were antagonized by D-(-)-2-amino-5-phosphonopentanoic acid, cis-4 (phosphonomethyl)piperidine-2-carboxylic acid, and 3-((R)-2-carboxypiperazin-4 yl)-propyl-1-phosphonic acid, all of which had slightly, but statistically significant, higher affinities (2.2-, 2.8-, and 5.5-fold, respectively) for the NR2A-containing receptor. Responses were also antagonized by the glycine-site antagonists 7-chlorokynurenic acid, 7-chloro-4-hydroxy-3-(3 phenoxy)phenylquinolin-2-(1H)-one, and (+/=)-4-(trans)-2-carboxy-5,7-dichloro-4 phenylaminocarbonylamino- 1,2,3,4- tetrahydroquinoline. The atypical NMDA antagonist ifenprodil showed the largest separation in functional affinity (IC50 values, 0.6 and 175 microM at NR2B- and NR2A-containing receptors, respectively). These experiments demonstrate the usefulness of permanently transfected L(tk-) cells for electrophysiological studies of recombinant NMDA receptor function and provide the first detailed functional pharmacological analysis of human NMDA receptor subtypes. PMID- 7476915 TI - Differential coupling of glucagon and beta-adrenergic receptors with the small and large forms of the stimulatory G protein. AB - Glucagon receptors (GRs) and beta-adrenergic receptors (beta-ARs) stimulate adenylate cyclase (AC) via Gs. The present study was performed to determine whether different cAMP-generating receptors share the same pool of Gs. In hepatocytes and liver plasma membranes from partially hepatectomized male rats, glucagon was more potent in stimulating AC than beta-adrenergic agonists, but the effects of glucagon and beta agonists on AC activity were not additive. This suggests that GRs and beta-ARs share the same pathway. Glucagon lowered the affinity of beta agonists for beta-ARs in the presence of GTP gamma S, whereas beta agonists had no effect on glucagon binding to GRs regardless of the presence or the absence of GTP gamma S. Therefore, the pool of Gs coupled to GRs appears to include that coupled to beta-ARs. The alpha subunit of Gs (Gs alpha) exists in small (Gs alpha-S) and large (Gs alpha-L) forms. Recently, with a new method that uses tryptic digestion, the G protein coupled to beta-ARs was identified as Gs-L in partially hepatectomized male rat livers because beta-adrenergic agonists promoted trypsinization of Gs alpha-L but not of Gs alpha-S. By contrast, the present study showed that glucagon enhanced the sensitivity of the two Gs alpha isoforms to trypsin in a concentration-dependent manner, indicating that GRs are coupled to both Gs alpha-S and Gs alpha-L. In conclusion, GRs share a common Gs-L with beta-ARs but are also coupled to another Gs, Gs-S, in partially hepatectomized male rat livers. PMID- 7476916 TI - Glutamine transaminase K is not a major cysteine S-conjugate beta-lyase of rat kidney mitochondria: evidence that a high-molecular weight enzyme fulfills this role. AB - Glutamine transaminase K (homodimer; M(r) of monomer approximately 45,000) is a major cysteine S-conjugate beta-lyase of rat kidney cytosol. Several cysteine S conjugates are known to cause kidney damage. Mitochondria are especially sensitive, and glutamine transaminase K activity is present in the mitochondrial fraction of rat kidneys. Therefore, it is possible that the mitochondrial form of glutamine transaminase K is a cysteine S-conjugate beta-lyase of the rat kidney and that this activity contributes to the mitochondrial damage. However, the literature contains conflicting data on this point. We obtained highly purified mitochondrial glutamine transaminase K and showed that it possesses little cysteine S-conjugate beta-lysae activity with S-(1,2-dichlorovinyl)-L-cysteine and S-(1,1,2,2-tetrafluoroethyl)-L-cysteine as substrates. Recently, a high molecular-weight cysteine S-conjugate beta-lyase (M(r) approximately 330,000) was shown to be present in the cytosol of rat kidney homogenates and partially purified. We present evidence that suggests that a similar high-molecular-weight enzyme is present in rat kidney mitochondria and that this protein may be a major cysteine S-conjugate beta-lyase of these organelles. PMID- 7476917 TI - Chronic ethanol consumption differentially alters the expression of gamma aminobutyric acidA receptor subunit mRNAs in rat cerebral cortex: competitive, quantitative reverse transcriptase-polymerase chain reaction analysis. AB - The molecular mechanisms that underlie ethanol dependence appear to involve alterations in GABAA receptor function and gene expression. In rat cerebral cortex, chronic exposure to ethanol alters many functional properties of GABAA receptors, including reduction of GABAA receptor-mediated chloride uptake. These functional alterations occur without a concomitant alteration in total receptor density or affinity. Previous investigations have shown that chronic ethanol exposure elicits alterations in mRNA and polypeptide levels for several abundant GABAA receptor subunits. For example, alpha 1 and alpha 2 subunit mRNA and polypeptide levels have been shown to decrease with chronic ethanol exposure. The present study was undertaken to further investigate the effects of chronic ethanol consumption on GABAA receptor subunit mRNA levels in rat cerebral cortex by using a competitive, quantitative reverse transcriptase-polymerase chain reaction assay that incorporates subunit-specific internal standards and allows for the absolute quantification of mRNA levels. We find that chronic ethanol consumption elicits a significant increase in alpha 4 subunit mRNA levels that is equal, in absolute amount, to a decrease in alpha 1 subunit mRNA levels. There is a small (30%) increase in gamma 2S but not gamma 2L subunit mRNA levels after chronic ethanol consumption. In addition, gamma 1 subunit mRNA levels are increased by 70%, whereas alpha 5, beta 1, beta 2, beta 3, gamma 3, and delta subunit mRNA levels do not change. We also reproduced results obtained previously by Northern blot analysis showing a 40% reduction in alpha 1 mRNA levels with no change in beta 2 subunit mRNA levels after chronic ethanol consumption. These results are consistent with the hypothesis that chronic ethanol consumption alters the function of GABAA receptors by eliciting changes in receptor subunit assembly. These changes may underlie the development of ethanol dependence. PMID- 7476918 TI - Molecular cloning and expression of multiple isoforms of human prostaglandin E receptor EP3 subtype generated by alternative messenger RNA splicing: multiple second messenger systems and tissue-specific distributions. AB - Five distinct cDNA clones encoding four different isoforms of human prostaglandin (PG) E receptor EP3 subtype were isolated from a human kidney cDNA library. Two cDNA clones differed only in their 3'-untranslated regions. The four isoforms, tentatively named EP3-I, EP3-II, EP3-III, and EP3-IV, which were generated by alternative mRNA splicing, had identical amino acid sequences except for their different carboxyl-terminal tails. Transfection experiments revealed that all the four isoforms show high binding affinities to PGE2, PGE1, and M&B28767, an EP3 specific agonist, whereas their downstream signaling pathways are divergent. M&B28767 increased cAMP concentrations in cells expressing EP3-II and EP3-IV, whereas it inhibited forskolin-induced cAMP accumulations in cells expressing all EP3 isoforms. M&B28767 also stimulated phosphoinositide turnover in cells expressing EP3-I and EP3-II. Northern blot analysis revealed that the EP3 gene is expressed in a wide variety of human tissues. The human EP3 mRNA was present most abundantly in the kidney, pancreas, and uterus. A substantial expression was also detected in the heart, liver, skeletal muscle, small intestine, colon, prostate, ovary, and testis. Furthermore, reverse transcription-polymerase chain reaction analysis demonstrated tissue-specific expressions of the five different EP3 mRNA species. The present study suggests the presence of the multiple systems of PGE2/EP3 isoforms and leads to the better understanding of its physiological and pathophysiological implications in humans. PMID- 7476920 TI - Activation of thromboxane and prostacyclin receptors elicits opposing effects on vascular smooth muscle cell growth and mitogen-activated protein kinase signaling cascades. AB - Thromboxane A2 stimulation of smooth muscle cells contributes to the development of vascular lesions after percutaneous transluminal coronary angioplasty. In view of this, we examined the signaling pathways stimulated by a thromboxane receptor agonist, U-46619, in cultures of rat aortic smooth muscle cells. Treatment of rat aortic smooth muscle cells with U-46619 induced cellular hypertrophy ([14C]leucine incorporation) without stimulating mitogenesis ([3H]thymidine incorporation). Analysis of signaling pathways elicited by U-46619 revealed enhanced tyrosine phosphorylation and increased enzymatic activity of mitogen activated protein (MAP) kinase (Erk2). U-46619 also activated signaling proteins upstream of p21-ras, inducing tyrosine phosphorylation on Shc and complex formation between Shc and growth factor receptor binding protein-2 (GRB2). Exposure of cells to a stable prostacyclin analogue, ciprostene calcium, attenuated U-46619-induced cellular hypertrophy and MAP kinase activity. Ciprostene treatment elevated cellular cAMP and inhibited U-46619-induced tyrosine phosphorylation on Shc and Shc/GRB2 complex formation. These results demonstrate that stimulation of thromboxane A2 and prostacyclin receptors have opposing effects on smooth muscle cell hypertrophy and the signaling pathways associated with this process. We conclude that inhibition of Shc/GRB2 complex formation and MAP kinase activity by ciprostene may contribute to its ability to limit restenosis injury. PMID- 7476919 TI - Regulation of delta FosB and FosB-like proteins by electroconvulsive seizure and cocaine treatments. AB - Previous work has shown that c-Fos and several Fos-like proteins or Fras (Fos related antigens) are induced acutely in brain in response to a wide variety of stimuli. In contrast, several stimuli induce apparently distinct Fos-like proteins, termed chronic Fras, after chronic administration. We show that delta FosB, a truncated splice variant of FosB, responds like the other acute Fras: it is induced rapidly and transiently in cerebral cortex after acute electroconvulsive seizure (ECS) and in striatum after acute cocaine but does not accumulate after chronic ECS or cocaine treatment. Although the chronic Fras are immunochemically related to delta FosB, they can be distinguished from delta FosB based on their temporal properties in that they are induced after chronic ECS and cocaine treatments only. Moreover, the chronic Fras and delta FosB can be distinguished by their migration patterns on one- and two-dimensional gel electrophoresis. The chronic Fras, therefore, appear to be novel FosB-related proteins. We also provide evidence that the chronic Fras heterodimerize primarily with Jun-D and Jun-B, as opposed to c-Jun. The possibility that AP-1 complexes containing the chronic Fras function as negative regulators of AP-1 mediated transcription is discussed. PMID- 7476921 TI - In vitro transcription of DNA containing 2-chloro-2'-deoxyadenosine monophosphate. AB - 2-Chloro-2'-deoxyadenosine (cladribine [CldAdo]) represents one of the most promising therapeutic agents for the treatment of pediatric leukemias and adult hairy cell leukemia. We examined whether CldAdo incorporation into DNA inhibited subsequent transcription in vitro using purified phage RNA polymerases. Control (Ade-containing) and 2-chloroadenine (ClAde)-substituted DNA strands that contained a RNA polymerase promoter sequence were synthesized by a modified asymmetric polymerase chain reaction. Complementary (+) and (-) strands were annealed, incubated with phage RNA polymerase, and analyzed with denaturing PAGE. When ClAde was present in both strands, the yield of full-length transcripts (approximately equal to 100 bases) was reduced by approximately equal to 90% relative to control DNA. Transcription was also reduced to a slightly lesser degree when substitutions occurred in only one of two strands. The observed low transcript levels on ClAde-containing DNA were due in part to the presence of the analogue within the promoter region. With gel shift binding assays, we demonstrated that RNA polymerase did not bind as well to ClAde-containing promoters. Polymerase/DNA complex formation was decreased by approximately equal to 80% compared with that on control unsubstituted promoters. In addition, on binding to the substituted promoter, RNA polymerase had an altered conformation that led to enhanced proteolytic clipping by endoproteinase Glu-C. Transcript sequence analysis indicated that SP6 RNA polymerase read through template ClAde residues with no apparent misincorporation into RNA. Our results provide insight into a novel effect of this nucleoside analogue that may explain its cytotoxicity in nondividing cells. PMID- 7476922 TI - Inhibition of splicing of wild-type and mutated luciferase-adenovirus pre-mRNAs by antisense oligonucleotides. AB - We report the construction, characterization, and use of luciferase reporters to test the ability of antisense oligonucleotides to inhibit RNA splicing. beta Globin and adenovirus introns were inserted into a luciferase cDNA, and luciferase expression was analyzed in transiently transfected cells. The adenovirus reporter expressed large amounts of luciferase, but two beta-globin constructs were inactive. RNA analyses determined that the beta-globin pre-mRNAs were not spliced. Mutagenesis of the beta-globin 5' splice site, branchpoint, and 3' splice site sequences to the adenovirus intron sequences promoted maximal splicing and luciferase activity; reciprocal changes in all three elements of the adenovirus intron eliminated luciferase activity. Wild-type and 3' splice site mutated adenovirus reporters were used to determine the ability of phosphorothioate deoxy and 2' methoxy oligonucleotides to inhibit splicing. RNase H activating oligodeoxynucleotides were better inhibitors of wild-type adenovirus expression than were 2' methoxy analogues. However, 2' methoxy oligonucleotides specific for the branchpoint were more effective inhibitors of splicing of adenovirus transcript containing the beta-globin branchpoint and 3' splice site. We suggest that pre-mRNAs with weak splice sites are potential targets for oligonucleotides that inhibit splicing by occupancy rather than cleavage of the transcripts. PMID- 7476923 TI - Pharmacology of the inhibitory glycine receptor: agonist and antagonist actions of amino acids and piperidine carboxylic acid compounds. AB - To define structure-activity relations for ligands binding to the inhibitory glycine receptor (GlyR), the agonistic and antagonistic properties of alpha- and beta-amino acids were analyzed at the recombinant human alpha 1 GlyR expressed in Xenopus oocytes. The agonistic activity of alpha-amino acids exhibited a marked stereoselectivity and was highly susceptible to substitutions at the C alpha atom. In contrast, alpha-amino acid antagonism was not enantiomer dependent and was influenced little by C alpha-atom substitutions. The beta-amino acids taurine, beta-aminobutyric acid (beta-ABA), and beta-aminoisobutyric acid (beta AIBA) are partial agonists at the GlyR. Low concentrations of these compounds competitively inhibited glycine responses, whereas higher concentrations elicited a significant membrane current. Nipecotic acid, which contains a trans-beta-amino acid configuration, behaved as purely competitive GlyR antagonist. Our data are consistent with the existence of a common binding site for all amino acid agonists and antagonists, at which the functional consequences of binding depend on the particular conformation a given ligand adopts within the binding pocket. In the case of beta-amino acids, the trans conformation appears to mediate antagonistic receptor binding, and the cis conformation appears to mediate agonistic receptor binding. This led us to propose that the partial agonist activity of a given beta-amino acid is determined by the relative mole fractions of the respective cis/trans conformers. PMID- 7476924 TI - Inhibition of succinate:ubiquinone reductase and decrease of ubiquinol in nephrotoxic cysteine S-conjugate-induced oxidative cell injury. AB - The role of complex II in the cellular protection against oxidative stress was investigated in freshly isolated rat renal proximal tubular cells (PTC) with the use of the nephrotoxin S-(1,2-dichlorovinyl)-L-cysteine (DCVC). DCVC caused oxidative stress in PTC as determined by flow cytometry with dihydrorhodamine 123; this fluorescent probe is readily oxidized by primary hydroperoxides such as those formed during lipid peroxidation. The oxidative stress could be prevented by inhibition of the beta-lyase-mediated formation and covalent binding to cellular macromolecules of reactive DCVC metabolites, with amino oxyacetic acid (AOA), or by the antioxidant N,N'-diphenyl-p-phenylenediamine. Both AOA and DPPD also prevented cell death. The DCVC-induced oxidative stress was associated with a decrease in the succinate:ubiquinone reductase (SQR) activity of complex II, whereas NADH:ubiquinone reductase activity of complex I remained unaffected. AOA prevented the effect on SQR activity, whereas N,N'-diphenyl-p-phenylenediamine did not. Inhibition of SQR activity with thenoyl trifluoracetone (TTFA) potentiated the DCVC-induced oxidative cell injury, suggesting the involvement of SQR activity in an antioxidant pathway. To investigate this in greater detail, PTC were treated with an inhibitor of cytochrome-c-oxidase, KCN, in a buffer containing glycine, which prevents cell death by KCN. Glycine did not affect cell death by DCVC. KCN prevented the DCVC-induced oxidative stress and cell death. KCN cytoprotection could be prevented by inhibition of SQR activity with oxaloacetate or TTFA, whereas inhibition of either complex I or III with rotenone and antimycin, respectively, did not prevent it. The effect of DCVC on complex II was associated with a decrease in the cellular amount of reduced ubiquinone (QH2); the KCN-mediated cytoprotection was related to a 60% increase of cellular QH2. Rotenone almost completely inhibited ubiquinone reduction even in the presence of KCN, whereas oxaloacetate in combination with KCN resulted in QH2 levels comparable to control. This suggests that the SQR activity by complex II rather than the cellular content of reduced ubiquinone (QH2) is important as a part of the cellular antioxidant machinery in the cyto-protection against oxidative stress. PMID- 7476925 TI - Zinc ions antagonize the inhibitory effect of aurothiomalate on glucocorticoid receptor function at physiological concentrations. AB - The water-soluble gold preparation aurothiomalate, which contains gold as Au(I), is frequently prescribed for patients with rheumatoid arthritis as a disease modifying agent. We report that aurothiomalate negatively modulates glucocorticoid hormone action; it represses the ligand- and DNA-binding activities and the transactivation function of the glucocorticoid receptor. We suggested the existence of endogenous titrating activities of Au(I) because otherwise administration of aurothiomalate to a patient with rheumatoid arthritis would be expected to result in peripheral insensitivity to glucocorticoids and worsen the patient's status. Focusing on metal ions that are present in vivo, we found that Zn(II) counteracts the inhibitory effect of Au(I) on glucocorticoid receptor function. This complementary effect of Zn(II) was observed at physiological concentrations. We suggest that Zn(II) preserves glucocorticoid receptor function in target tissues and maintains hormone responsiveness, even with chrysotherapy. PMID- 7476926 TI - Interactions among GYKI-52466, cyclothiazide, and aniracetam at recombinant AMPA and kainate receptors. AB - We examined the actions of cyclothiazide, aniracetam, and 1-(4-aminophenyl)-4 methyl-7,8-methylenedioxy-5H-2,3-benzodiazepine (GYKI-52466) on recombinant alpha amino-3-hydroxy-5-methyl-4-isoxazole propionate (AMPA) and kainate receptors. Receptors expressed in Xenopus oocytes or human embryonic kidney 293 cells were characterized using voltage and patch-clamp electrophysiology. Aniracetam and cyclothiazide potentiated AMPA receptor currents by slowing or blocking desensitization. Cyclothiazide was more potent at receptors consisting of flip subunits compared with receptors consisting of flop subunits, whereas aniracetam appeared to be more efficacious at flop receptors. The potency of GYKI-52466 did not differ in heteromeric flip or flop containing AMPA receptors, but GYKI-52466 was less potent at homomeric GluRAi and GluRDi receptors. At heteromeric AMPA receptors, 50 microM cyclothiazide increased the IC50 value for GYKI-52466 significantly. The increase was largest in GluRBi/Di receptors where the IC50 value shifted from 21.9 microM (95% confidence interval, 12.0-39.8 microM) to 126 microM (95% confidence interval, 72.4-214 microM) in the presence of cyclothiazide. In contrast, 100 microM GYKI-52466 did not alter the EC50 of cyclothiazide at GluRBi/Di receptors nor did it markedly change the maximal potentiation induced by cyclothiazide. At GluRBi/Di receptors transiently expressed in human embryonic kidney 293 cells, 30 microM GYKI-52466 inhibited the steady state and the peak current evoked by 300 microns L-glutamate to the same extent (34.5 +/- 12% and 27.3 +/- 13.0%, respectively; five experiments), and GYKI-52466 did not alter the apparent rate of desensitization (tau = 15.7 +/- 4.7 and 17.5 +/- 8.3 msec in the absence and presence of GYKI-52466, respectively; five experiments). GYKI-52466 inhibited L-glutamate currents in the presence and absence of 10 microM cyclothiazide, but GYKI-52466 never restored the desensitization that was blocked by cyclothiazide. Furthermore, GYKI-52466 inhibited L-glutamate currents mediated by homomeric Glu6 receptors, which are not potentiated by cyclothiazide. Our data suggest that the effect of cyclothiazide on the affinity of GYKI-52466 for its binding site is allosteric and that the positive modulatory effect of cyclothiazide and the negative modulatory effect of GYKI-52466 result from binding to separate sites on recombinant subunits. PMID- 7476927 TI - Agonists and antagonists differentially regulate the high affinity state of the D2L receptor in human embryonic kidney 293 cells. AB - Studies with radiolabeled antagonists have revealed that both agonists and antagonists induce up-regulation of D2 dopamine receptors in cells transfected to express D2L or D2S receptors. The regulation induced by agonists, but not antagonists, was synergistic with cAMP analogues, and differences in the time courses of the effects of agonists and antagonists have been observed. These findings have been extended by using a radiolabeled agonist to investigate agonist- and antagonist-induced regulation of the high affinity state of the D2L dopamine receptor in transfected HEK 293 cells. Exposure to agonists decreased the proportion of receptors in the high affinity, agonist-preferring state. Exposure to antagonists, however, led to an increase in the density of receptors with a high affinity for agonists. The effects of both agonists and antagonists on the agonist-preferring receptors occurred without a lag and were time and dose dependent. Inhibition of forskolin-stimulated cAMP accumulation by agonists was not affected by exposure of the cells to the antagonist (-)-sulpiride. Desensitization was seen after exposing cells to the agonist quinpirole for 1.5 hr, suggesting that the rapid loss of high affinity binding sites represents an uncoupling of the receptor from the G protein that mediates inhibition of adenylyl cyclase. Pretreatment of cells with the protein synthesis inhibitor cycloheximide did not block the quinpirole-induced loss of receptors with a high affinity for agonists. The effect of (-)-sulpiride on high affinity binding sites was blocked by cycloheximide, but only after incubation of cells for sufficient time to induce an increase in the total number of receptors. After incubation of cells with (-)-sulpiride for a short time, the increase in the number of receptors with a high affinity for agonists was unaffected by cycloheximide. These results suggest that the increase in agonist binding after brief exposure to an antagonist is due to interactions of the receptor with one or more G proteins that are not coupled to inhibition of adenylyl cyclase, whereas the increase in agonist binding at later time points is associated with the antagonist-induced up-regulation. PMID- 7476928 TI - Assessment of stress gene mRNAs (HSP-27, 60 and 70) in obstructed rabbit urinary bladder using a semi-quantitative RT-PCR method. AB - Stress proteins (HSPs) participate in the cellular response to various stresses including hyperthermia, hypoxia and injury. A previous work using northern blot analysis demonstrated increased expression of stress protein 70 (HSP-70) in rabbit bladder tissue subjected to partial outlet obstruction. In order to determine if the increased expression was specific for HSP-70 or, alternatively, indicated a generalized stress protein response, a modified quantitative RT-PCR technique was used to quantitate HSP mRNAs (HSP-27, 60, and 70) in normal and obstructed rabbit urinary bladder tissues. The results show the following: 1) The modified semi-quantitative RT-PCR is a sensitive and reproducible technique for detecting mRNA in bladder tissue. 2) Constitutive levels of HSP-27, HSP-60, and HSP-70 mRNAs were detected in control bladder tissues; the relative signal intensity was highest for HSP-70 and lowest for HSP-27. 3) A transient increase in HSP mRNAs was observed after obstruction; the mRNAs of HSP-27, 60 and 70 increased 4.3-, 5.6-, and 2.4-fold, respectively, at 24 h following obstruction, then gradually returned to control levels by the end of one week post-obstruction and remained stable up to 14 days post-obstruction. These data indicate that the modified quantitative RT-PCR is a useful technique for detecting mRNA in bladder tissue; the stress response which occurs in rabbit urinary bladder tissue following partial outlet obstruction is a general phenomenon. PMID- 7476929 TI - Amino acid uptake in skeletal muscle of rats bearing the Yoshida AH-130 ascites hepatoma. AB - Rats bearing the Yoshida AH-130 ascites hepatoma show decreased activity of neutral amino acid transport in skeletal muscle measured in vivo as the tissue accumulation of the analogue alpha-amino [1-14C]isobutyrate (AIB). The decreased accumulation of AIB observed is not merely a consequence of the hypoinsulinaemia present in these animals (as a result of tumour burden) since in vitro experiments carried out using incubations of isolated soleus muscles also showed a decreased uptake of neutral amino acids. In these preparations the addition of insulin results in similar increases in uptake both in the pair-fed controls and the tumour-bearing animals, thus suggesting similar insulin sensitivities. The decrease in amino acid uptake in soleus muscle is associated with a decrease in the activity of system A, while systems L and ASC show no particular changes as a result of the tumour growth. The kinetic characterisation of system A in the Yoshida-bearing rats shows a decrease in Vmax together with a decrease in KM in relation with the pair-fed animals. PMID- 7476930 TI - Enhanced expression of mRNA for insulin-like growth factor-1 in post-burn hypertrophic scar tissue and its fibrogenic role by dermal fibroblasts. AB - Hypertrophic scarring (HSc) which frequently develops in patients following severe thermal injury is characterized by accumulation of extracellular matrix (ECM) proteins including type I and type III collagen. In this study, we examined the presence and quantity of IGF-1 mRNA transcripts in post-burn HSc. The results of dot blot experiments showed a 77.5% (100 +/- 8.15 vs 177.5 +/- 19, p < 0.01) increase in expression of IGF-1 IIIRNA in HSc tissue relative to normal dermis obtained from the same patients. A Northern blot analysis confirmed the specificity of the IGF-1 cDNA. This cDNA visualized four different transcripts with apparent sizes of 7.0, 3.9, 1.8 and 1.0 kb, similar to those previously reported. The possible fibrogenic role of IGF-1 was examined by analyzing the effect of this growth factor on the expression of mRNA for the pro alpha 1(I) chain of type I procollagen and the pro alpha 1(III) chain of type III procollagen in dermal fibroblasts. IGF-1 increased the expression of these transcripts as early as 6 h and the effect was maximal at 24 h. Quantitative analysis by densitometry showed a 149 and 166% increase in pro alpha 1(I) and pro alpha 1(III) mRNA after 24 h of IGF-1 treatment, respectively. This effect seems to be specific as the abundance of mRNA for the pro alpha 2(I) chain of type I procollagen or TIMP-II was unchanged.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7476931 TI - Effect of nuclear Ca2+ uptake inhibitors on Ca(2+)-activated DNA fragmentation in rat liver nuclei. AB - The effect of nuclear Ca2+ uptake inhibitors on the Ca(2+)-activated DNA fragmentation in rat liver nuclei was investigated. The addition of Ca2+ (40 microM) into the reaction mixture containing liver nuclei in the presence of 2.0 mM ATP caused a remarkable increase in nuclear DNA fragmentation. This Ca(2+) activated DNA fragmentation was not seen in the absence of ATP, because nuclear Ca2+ uptake is not initiated without ATP addition. Moreover, the presence of various reagents (10 microM arachidonic acid, 2.0 mM NAD+, 10 microM zinc sulfate and 0.2 mM N-ethylmaleimide), which could inhibit Ca(2+)-ATPase activity and Ca2+ uptake in the nuclei, produced a significant inhibition of the Ca(2+)-activated DNA fragmentation in the nuclei. The results show that the Ca(2+)-activated DNA fragmentation is involved in the uptake of Ca2+ by the nuclei, suggesting a role of Ca2+ transport system in the regulation of liver nuclear functions. PMID- 7476932 TI - Effect of hydralazine on myocardial plasma membrane fatty acid binding protein (PM-FABP) during diabetes mellitus. AB - Chronic treatment with the antihypertensive drug hydralazine did not affect the hyperglycemic state of streptozotocin (STZ)-diabetic rats but did prevent the serum hyperlipidemia that is synonymous with these diabetic animals. After 6 weeks, untreated STZ-diabetic rats exhibited a 659% increase in serum triglycerides and 292% increase in serum cholesterol versus age-matched non diabetic rats. Hydralazine-treated STZ-diabetic rats had serum triglyceride and cholesterol levels that did not differ from controls. Myocytes from control rats showed a preference for binding of the unsaturated fatty acid analog cis parinaric acid vs the saturated fatty acid analog trans-parinaric acid. This preference was not altered in STZ-diabetic rat myocytes; hydralazine-treatment of STZ-diabetic rats also showed no change in fatty acid preference. STZ-diabetes caused a decrease in the affinity (Kd) for the trans, but not the cis-parinaric acid. However, total binding of both analogs was increased in STZ-diabetes. Hydralazine treatment of STZ-diabetic rats resulted in even greater total binding of both analogs. Affinity for the trans analog was further decreased in these hydralazine-treated rats, but the affinity for the cis analog was increased beyond control animals. These results suggest that the diabetic state influences the binding characteristics of the myocardial PM-FABP and that hydralazine, while reducing serum lipids in diabetes, has complex effects on the fatty acid binding by this protein. PMID- 7476933 TI - Thiol modulation of TNF alpha and IL-1 induced MnSOD gene expression and activation of NF-kappa B. AB - TNF alpha and IL-1 each can activate NF-kappa B and induce gene expression of manganese superoxide dismutase (MnSOD), a mitochondrial matrix enzyme which can provide critical protection against hyperoxic lung injury. The regulation of MnSOD gene expression is not well understood. Since redox status can modulate NF kappa B and potential kappa B site(s) exist in the MnSOD promoter, the effect of thiols (including NAC, DTT and 2-ME) on TNF alpha and IL-1 induced activation of NF-kappa B and MnSOD gene expression was investigated. Activation of NF-kB and increased MnSOD expression were potentiated by thiol reducing agents. In contrast, thiol oxidizing or alkylating agents inhibited both NF-kappa B activation and elevated MnSOD expression in response to TNF alpha or IL-1. Since protease inhibitors TPCK and TLCK can inhibit NF-kappa activation, we also investigated the effect of these compounds on MnSOD expression and NF-kappa B activation. TPCK and TLCK each inhibited MnSOD gene expression and NF-kappa B activation. Since the MnSOD promoter also contains an AP-1 binding site, the effect of thiols and thiol modifying agents on AP-1 activation was investigated. Thiols had no consistent effect on AP-1 activation. Likewise, some of the thiol modifying compounds inhibited AP-1 activation by TNF alpha or IL-1, whereas others did not. Since diverse agents had similar effects on activation of NF kappa B and MnSOD gene expression, we have demonstrated that activation of NF kappa B and MnSOD gene expression are closely associated and that reduced sulfhydryl groups are required for cytokine mediation of both processes. PMID- 7476934 TI - Protective effect of seminal plasma proteins on the degradation of ascorbic acid. AB - The objectives of this study were to determine ascorbic acid stability and its effect on antiproteinase activity of seminal plasma in the presence of an oxidant. Effect of seminal plasma, and additives: glutathione, albumin, hydrogen peroxide and Tris buffer, on ascorbic acid degradation was investigated by UV absorbance. Antiproteinase against trypsin amidase activity was measured spectrophotometrically using N-benzoyl-DL-arginine-p-nitroanilide (BAPNA) as substrate. Ascorbic acid was destroyed much more rapidly with the addition of hydrogen peroxide than in Tris buffer at pH 8.2 alone. Seminal plasma protected ascorbic acid more efficiently than glutathione and albumin alone. The protective effect of seminal plasma on ascorbic acid degradation may closely relate to the function of ascorbic acid in reproductive system of scurvy-prone animals including teleost fish. Within the range of 1-8 mM concentrations, ascorbic acid had a pro-oxidant action on seminal plasma antiproteinase activity in vitro when they were incubated with hydrogen peroxide. PMID- 7476935 TI - Characterization of regucalcin effect on proteolytic activity in rat liver cytosol: relation to cysteinyl-proteases. AB - The increasing effect of regucalcin, isolated from rat liver cytosol, on neutral proteolytic activity in the hepatic cytosol was characterized. The proteolytic activity was markedly elevated by the addition of regucalcin (0.1-0.5 microM) in the absence of Ca2+. This increase was not significantly altered by the presence of diisopropylfluorophosphate (DPF; 2.5 mM)--although DFP caused a significant decrease in the proteolytic activity. Regucalcin (0.25 microM) additively enhanced the dithiothreitol (DTT; 1.0 mM)--increased proteolytic activity, while the regucalcin or DTT effect was completely abolished by NEM (5 mM), indicating that regucalcin may act on the SH group in proteases. Also, regucalcin (0.25 microM) enhanced the effect of Ca2+ (10 microM) increasing liver proteolytic activity, suggesting that regucalcin does not influence on the active sites for Ca2+ in proteases. Moreover, the proteolytic activity of regucalcin (0.25 microM) was significantly decreased by the presence of calpastatin (24 micrograms/ml), an inhibitor of Ca(2+)-activated neutral protease (calpain). Now, regucalcin (0.25 microM) increased about 7-fold the activity of m-calpain isolated from rabbit skeletal muscle. These observations demonstrate that regucalcin directly activates cysteinyl-proteases. Regucalcin may have a role as a potent proteolytic activator in the cytoplasm of liver cells. PMID- 7476936 TI - Partial restoration of impaired alpha 1-adrenergic responsiveness in parotid cells of aged rats by S-adenosylmethionine treatment. AB - The age related decrease in alpha 1-adrenergic stimulated inositol 1, 4, 5 trisphosphate (IP3) production in parotid cells of aged rats can be partially restored by treatment with S-adenosylmethionine (SAM). This effect is completely blocked by S-adenosyl homocysteine (SAH) and occurs in association with an increase in the conversion of phosphatidylethanolamine to phosphatidylcholine and a decrease in membrane viscosity. In contrast, SAM treatment actually inhibits stimulated IP3 production in cells of young rats. The membrane viscosity of these cells is lower than that of those from aged rats. Although conversion of phosphatidylethanolamine to phosphatidylcholine is enhanced, no further decrease in membrane viscosity is elicited in young cell preparations. These findings suggest that age changes in the membrane environment may result in impaired alpha 1-adrenergic signal transduction and that such alterations may be at least partially reversible by SAM treatment. PMID- 7476937 TI - The role of tau phosphorylation in transfected COS-1 cells. AB - Tau cDNAs from each of the six human isoforms were transfected into COS-1 cells and, in every case, more than one peptide was observed. The diversity of expressed isoforms was due to different levels of tau phosphorylation. Tau phosphorylation results in a decrease of the protein electrophoretic mobility. The major contribution to this mobility shift is due to the phosphorylation at the at the C-terminus of the molecule, as inferred from the expression of tau fragments. Phosphorylation takes place in some of the sites modified in neural cells and in the basis of AD patients. Copolymerization studies indicate that the level of phosphorylation, as well as the localization of the modified residues, may affect the binding of the protein to microtubules. These results indicate that phosphorylation regulates tau function inside the cell. PMID- 7476938 TI - Regulation of the calcium slow channel by cyclic GMP dependent protein kinase in chick heart cells. AB - In order to assess the interaction between the cAMP-dependent and the cGMP dependent phosphorylation pathways on the slow Ca2+ current (ICa(L)), whole-cell voltage-clamp experiments were conducted on embryonic chick heart cells. Addition of 8Br-cGMP to the bath solution reduced the basal (unstimulated) ICa(L). Intracellular application of the catalytic subunit of PK-A (PK-A(cat); 1.5 microM) via the patch pipette rapidly potentiated ICa(L) by 215 +/- 16%) (n = 4); subsequent addition of 1 mM 8Br-cGMP to the bath reduced the amplitude of ICa(L) towards the initial control values (123 +/- 29%). Intracellular application of PK G (25 nM pre-activated by 10(-7) M cGMP), rapidly inhibited the basal ICa(L) by 64 +/- 6% (n = 8). Heat-denatured PK-G was ineffective. Subsequent additions of relatively high concentrations of 8Br-cAMP (1 mM) or isoproterenol (ISO, 1-10 microM) did not significantly remove the PK-G blockade of ICa(L). The results of the present study suggest that: (a) 8Br-cGMP can inhibit the basal or stimulated (by PK-A(cat)) ICa(L) in embryonic chick myocardial cells. (b) PK-G applied intracellularly inhibits the basal ICa(L). PMID- 7476940 TI - [Multiple sclerosis: molecular and cellular mechanisms]. AB - Multiple sclerosis (MS) is a chronic central nervous system disease of considerable medical and social impact. It is characterized by destruction of the myelin, the axon proteolipid sheath, or demyelination. While the etiology of MS remains unknown, one of the most well-grounded theories of its pathogenesis postulates that immunomediated inflammatory processes play the main role in myelin damage. The leading role in the autoimmune disturbance development belongs to T-cell system, however, B-cells also participate in the pathological process. Both genetical predisposition and environmental influence are involved in MS development. Correlations were found between MS and numerous environmental factors, including ecology and different infectious agents. However, no single environmental factor and no single infection was confirmed to be the primary cause of MS. The predisposition to MS seems to depend on several genes. Alleles and haplotypes of HLA genes which are the main human immune-response genes are undoubtedly associated with MS. Serological methods have shown weak association of MS with A3 and B7 loci of HLA class I. Stronger association was found for HLA class II haplotype specified to DR2(DR15), DQ6(DQ1) in serology typing nomenclature or DRB1*1501, DQA1*0102, DQB1*0602 in sequence-based genotyping terminology. Besides, MS was found to be associated with alleles of genes of T cell receptors, cytokines, myelin components and some others, although these results are sometimes contradictory. The analysis of genetical predisposition factors and of possible mechanisms of their involvement in demyelination process on molecular and cellular levels should enlighten the MS pathogenesis and provide new ways of medical treatment and prevention of MS. PMID- 7476939 TI - Lipopolysaccharide (LPS) increases the in vivo oxidation of branched-chain amino acids in the rat: a cytokine-mediated effect. AB - Septic rats (as induced by cecal puncture and ligation) showed an increased rate of in vivo leucine oxidation as measured from the formation of 14CO2 from an intravenously injected [1-14C]leucine tracer dose. Acute lipopolysaccharide (LPS) administration (1 mg/kg) to rats caused a similar effect on the rate of in vivo leucine oxidation. Additionally, both tumour necrosis factor-alpha (TNF) and interleukin-1-alpha (IL-1), in an acute dose of 100 micrograms/kg, also increased the rate of the oxidation of the amino acid, although only IL-1 caused a similar increase to that observed following LPS. The observed increased leucine oxidation was related to lower leucine concentrations both in LPS- and cytokine-treated rats. Important decreases were also observed in the other branched-chain amino acids (valine and isoleucine) in the LPS- and IL-1-treated animals. Isolated incubated muscles from TNF- and IL-1-treated rats did not show any changes in the rate of leucine utilization, thus suggesting that the mechanism by which the cytokines stimulate whole-body leucine oxidation is not based on an increase in the activity of the enzymatic machinery responsible for leucine oxidation. Additionally, glucocorticoids do not seem to mediate the enhanced in vivo oxidation of the amino acid since, although they are increased by both LPS and cytokines, treatment of the animals with RU486 (a glucocorticoid antagonist) was not able to suppress the effects of the cytokine on in vivo leucine oxidation. PMID- 7476941 TI - [Structure and functional similarity of yeast Sup35p and Ure2p proteins to mammalian prions]. AB - The results of studies of yeast cytoplasmically-inherited determinants [psi] and [URE3] are summarized. The existence of tandem amino acid repeats in N-terminal regions of yeast Sup35p protein and in prions of higher eukaryotes is shown. The prion-like properties of yeast Sup35p and Ure2p proteins and a role of tandem amino acid repeats localized in N-terminal region of Sup35p protein in inheritance of [psi] determinants is discussed. The suggestion is made that the [psi+] status of yeast cell depends on the specific conformation of the N terminal domain of Sup35p and that this protein can induce the specific conformational state of its N-terminal domain on newly synthesized Sup35p molecules via protein-protein interaction, thus representing a molecular basis of inheritance of [psi+] determinant. Other proteins containing amino acid repeats of similar type are considered and the suggestion is made that some of these proteins may show prion-like behavior. PMID- 7476942 TI - [Characteristics of hybrid genes coding functionally-active secretory metalloproteinases from bacilli]. AB - A set of different hybrid genes encoding functionally active enzymes was obtained by homologous recombination between the fragments of related Bacillus amyloliquefaciens and Bacillus brevis metalloprotease genes cloned in plasmid vector in tandem orientation. The nucleotide sequences of hybrid genes were analyzed. It was demonstrated that the presence even of short homologous regions is sufficient for effective recombination in Bacillus cells. PMID- 7476943 TI - [A new class of RNP particles containing small RNA homologous to short dispersed DNA repetitive sequences]. AB - Here we show that small RNAs homologous to short interspersed repetitive DNA sequences: ID, B1, B2--in rat cells and Alu in human cells are complexed with specific proteins to form small nuclear and cytoplasmic RNP particles (alpha RNP) with common properties. alpha-RNP differ from other ribonucleoproteins by composition and properties. alpha RNA molecules are apparently transcribed by RNA polymerase III. alpha-RNAs are capable of stable antisense hybridization with specific messenger RNAs. Expression of alpha-RNA is specifically regulated by gene regulatory factors. The data obtained support the suggestion that alpha-RNA may belong to the group of regulatory eukaryotic RNAs and that alpha-RNP might be involved in the coordinative control of the expression of the sets of genes with SINE-homologous sequences in regulatory regions. PMID- 7476945 TI - [Plan for finding homologies in nucleotide sequence databases using preliminarily calculated sequence samples]. AB - A scheme of fast similarity search of nucleotide sequences is suggested based on sequence imaging, which results in chunks of information much less than original sequence but more specialized for comparison. Three methods were developed using three different imaging functions. The first is based on identity of local sites of up to twelve nucleotides, the second is based on statistical homology of local 42 nucleotide fragments, and the third is based on the homology of 100-150 nucleotide fragments and models the comparison of restriction maps. Each of them requires the library of sequence images. The total size of such a library is less than the size of sequences stored in compressed form. The sequences are aligned allowing local homology searches. The method reduces total time for a similarity search about 100-fold. The programs can be easily included in any software, which allows user to define his own set of sequences. One of the programs is implemented within DNA-SUN software and is used in Institute of Molecular Genetics and Institute of Molecular Biology. PMID- 7476944 TI - [Study of the structure-function organization of the variola virus genome. IV. Sequencing and analysis of the nucleotide sequence of the right terminus of the India-1967 strain genome]. AB - Sequencing and computer analysis of the variola major virus strain India-1967 (VAR-IND) genome segment (53,018 bp) from the right terminal region have been carried out. Fifty nine potential open reading frames (ORFs) of over 60 amino acid residues have been identified. Structure-function organization of VAR-IND DNA segment under study was compared with the previously reported sequences from the analogous genomic regions of vaccinia virus strains Copenhagen (VAC-COP) and Western Reserve (VAC-WR) and variola virus strain Harvey (VAR-HAR). Multiple distinctions in the genetic map of VAR-IND from VAC-COP and VAC-WR have been revealed along with the high similarity to the corresponding VAR-HAR segment. Possible functions of the predicted viral proteins and the effect of their differences on the features of orthopoxviruses are discussed. PMID- 7476947 TI - [Characteristics of the RFLP marker of the D13S25 locus on human chromosome 13: mapping of SspI sites in the pH2-42 probe]. AB - Analysis of probe pH2-42 of locus D13S25 in genomic DNA in Moscow population has been made by PCR-RFLP using restriction endonuclease SspI. Localization of additional SspI-site at position 1240 (which has been revealed earlier by sequencing of pH2-42) was confirmed. This SspI restriction site was present in all representatives of the population, thus being not polymorphic. Consequently establishment of identity of additional site at position 1240 with polymorphic SspI site in D13S25 studied in previous investigations, requires additional data from another populations. Possible localization of polymorphic SspI-site in locus D13S25 is discussed. PMID- 7476946 TI - [Recombination properties of the modified Pseudomonas aeruginosa RecA protein]. AB - Gene recA from Pseudomonas aeruginosa was cloned into pUC19 vector under lacZ promoter. The expressed protein appeared to be modified, the aminoterminal part of deduced amino acid sequence of the RecAPa protein was found elongated by a polypeptide of 10 amino acids. The modified protein named RecA*Pa completely replaces RecAEc from E. coli in vivo recombination. In vitro RecA*Pa promotes the homologous strand transfer from a short linear duplex DNA fragment (346 bp) into circular single-stranded DNA (8196 n) being 5 times more active than RecAEc. However, when the length of dsDNA increased the difference between two proteins becomes negligible. To understand the reasons, some properties of RecA*Pa and RecAEc were compared. The former was shown to be more active both in binding to ssDNA in ssDNA-dependent ATP hydrolysis. The Rec*Pa protein showed also a high affinity to dsDNA, even at a physiological pH which is known to be unfavorable for RecAEc/dsDNA binding. However, both proteins equally catalyzed the dsDNA dependent ATP hydrolysis; we suggest that this is crucial for a full-length DNA strand transfer recombination reaction. PMID- 7476948 TI - [Phylogenetic relationships between 16 species of Lake Baikal sculpin fish based on a nucleotide sequence analysis of a mitochondrial DNA fragment of the cytochrome b gene]. AB - The fragment of mitochondrial cytochrome b gene from 16 species of Baikalian sculpins belonging to three families--Cottidae, Comephoridae and Abyssocottidae- was amplified by polymerase chain reaction and sequenced. Phylogenetic trees were built from data obtained. Some disconformity were noted between systematical and phylogenetic notions of today and obtained results. For example, branch lengths among some Cottidae species are larger than among any different family species. Moreover, schemes indicate separate position of Limnocottus euristomus from Limnocottus genus species. Phylogenetic trees confirm earlier conclusions, about recent--in geological scales of times--origin of baikalian Cottoidei (2-2.5 Myr). PMID- 7476949 TI - [Cloning an extended palindromic sequence of Tetrahymena pyriformis ribosomal DNA]. AB - Simple and effective method for isolation of native copies of ribosomal DNA (rDNA) of T. pyriformis by pulse field gel electrophoresis is suggested. Cloning of long palindrome sequence of rDNA from T. pyriformis in plasmid is described. PMID- 7476950 TI - [Heterogeneity of the structural organization of the chromatin domain including the dihydrofolate reductase gene in Chinese hamster ovary cell culture]. AB - We investigated histone-DNA interactions in chromatin of amplified about 1000 fold dihydrofolate reductase (DHFR) domain in methotrexate resistant Chinese hamster ovary cell line CHOC 400. We explored chromatin structure of DHFR-gene and extended region lying 3'-downstream of the DHFR-gene, which includes two zones of DNA replication initiation, Ori-beta and Ori-gamma, and matrix attachment region (MAR). Using the method of hybridization with "protein images" we found that the yield of both core histones and histone H1 cross-linked with DNA in 5'-region of the DHFR-gene, in Ori-gamma and regions flanked from either side MAR is significantly decreased in comparison with 3'-transcribed region of the DHFR-gene, Ori-beta and MAR. It was shown previously that decreased yield of histones cross-linked with DNA is typical for 5'-region of actively transcribed genes. So, histone-DNA interactions in chromatin of the 5'-region of the house keeping and moderately transcribed DHFR-gene, Ori-gamma and regions which flank MAR resemble those in 5'-regions of actively transcribed genes. Significant differences in amount of histones cross-linked with DNA in various parts of the DHFR-domain testify to high level of heterogeneity of chromatin structure in the domain. PMID- 7476951 TI - [Theory of cooperative transition of DNA complexes with multimodal ligands]. AB - The theory of the cooperative transition of DNA.ligand complexes have been developed, in which a model was implied where the multimodal ligands simultaneously interacted with DNA. Obtained formula express the dependence of the experimentally estimated values of the changes of transition point and width of transition on the concentration of the ligands. These expressions make possible to obtain the thermodynamic parameters of the interaction of the multimodal ligands with DNA (binding constants, number of base pairs corresponding to one binding site of DNA etc.) by comparing the theoretical and the experimental data. PMID- 7476953 TI - [Visualization of DNA segments, interacting with reactive oligonucleotide derivatives in interphase nuclei and metaphase chromosomes]. AB - Reaction of (pdT)16 derivatives, bearing 4-(N-2-chloroethyl-N methylamino)benzylphosphamide group on its 5' end and biotin on its 3' end with DNA in interphase nuclei and metaphase chromosomes has been investigated by fluorescence and electron microscopy. The result obtained evidence that in interphase nuclei DNA in active chromatin (nucleolus) is the most available for specific modification. In metaphase chromosomes the modified DNA regions are situated on the surface of chromosome. PMID- 7476952 TI - [Interaction of synthetic ligands containing N,4-disubstituted mono- and diphthalimides with DNA]. AB - Nevel synthetic DNA binding ligands containing N,4-disubstituted mono- and diphthalimide fragments and carrying positively charged groups at both ends of the molecules were characterized by various physiochemical methods. All the ligands were shown to have fluorescence maximum at about 460 nm for excitation at 340 nm. Interaction of these compounds with DNA and synthetic polydeoxyribonucleotides was studied using spectrophotometry, CD and fluorescence measurements. It was shown, that the ligand containing phthalimide fragment connected by flexible chain to dipyrrolcarboxamide fragment, binds to DNA narrow groove covering 5 base pairs. This type of complex formation was found to be followed by increasing of CD band at 280 nm and increasing in fluorescence intensity. If the ligand backbone contains only phthalimide fragments there is no binding in the DNA narrow groove. In this case the weakly fluorescent outside binding type complex is formed, which is characterized by the binding of one ligand molecule with two base pairs. PMID- 7476954 TI - [DNA-methylase activity in human cardiac muscle. Association of DNA methylase activity with actin protein fractions]. AB - The column isoelectrofocusing activity of the nuclear extracts of the human cardiac muscle has revealed at pH 3.5-8.2 5 peaks of DNA-methylase. When one of these peaks (II) was analyzed by the two-dimensional gel electrophoresis 6 proteins (10, 25, 35, 43, 67 and 120 kDa) were separated. 43 kDa protein had electrophoretic properties similar to actins and was able to methylate cytosine in the DNA molecules. The comparative computer analysis of the primary structure of human actins and several bacterial DNA-methylases has shown the homology of the extensive fragments of these molecules. PMID- 7476955 TI - [Do catalytically active antibodies exist in healthy people? (Protein kinase activity of sIgA antibodies from human milk)]. AB - The fraction of secretory immunoglobulin A (sIgA) from the milk of healthy mothers was purified by sequential affinity chromatography on protein A-sepharose (in the presence of 1% triton x100), adsorbent Toyopearl HW-55 (gel-filtration), DEAE cellulose (separation of IgG and IgA antibodies), affinity sorbents with immobilized ATP and casein. The protein obtained corresponded to sIgA antibodies according to all known criteria and did not contain any protein contaminations. The ability of sIgA to phosphorylate selectively serine residues of casein (not histones) in the presence of [gamma-32P]ATP was shown. Purified kinase activity was stable at acid shock (pH 2.3), strongly interacted with immobilized antibodies against H-chain of sIgA and eluted from the sorbent with the peak corresponding to sIgA antibodies. The complex of sIgA and ATP was stable enough at the conditions of gel-filtration. Affinity modification of sIgA by chemically reactive analogs of ATP resulted in preferential modification of its light chain (2-3 mole reagent per mole of dimer form). In the condition of oligomer dissociation ATP-sepharose sorbed only the light chains of sIgA. sIgA have optimal conditions for phosphorylating activity different from those of known protein kinases. We suppose that sIgA antibodies with kinase activity are a first example of sIgA antibodies with catalytic activity. For the first time the possibility of existence of natural abzymes in a healthy human was shown. These abzymes catalyze the reaction of synthesis but not substrate degradation reaction. PMID- 7476956 TI - [A bank of protein family images Prof-Image for rapid determination of possible functions of random amino acid sequences]. AB - A bank of images of protein families (PROF IMAGE bank) is developed using early published method and amino acid sequences of 24-th release SWISS-PROT bank (27752 sequences). The method relies on physical-chemical and structural properties of amino acids and on the choice of fragments of protein families for the most discriminate distinction of amino acid sequences of a family from representatives of other families or random sequences. Specifications of the algorithms of building the images, principles of amino acid sequences selection to form protein families, the structure of the bank, and characteristics of images of 163 protein families are described. The data are illustrated by the image of alpha-interferon precursors family. The results of the images comparison with all proteins of the SWISS-PROT bank and ways to use the PROF IMAGE bank for determination of possible functions of amino acid sequences are discussed. PMID- 7476958 TI - [Use of thermostable DNA polymerase from Thermus thermophilus KTP in a combined reverse transcription and amplification reaction of detecting interleukin 2alpha RNA and determining expression of the multidrug resistance gene (MDR-1)]. AB - According to our data native Tth DNA polymerase displays higher reverse transcription activity than Taq DNA polymerase. This allows one to use Tth DNA polymerase in the complete reaction of reverse transcription and amplification (RT/PCR). We used this enzyme to synthesize the interleukine (IL-2 alpha) RNA template synthesized by the RT/PCR method in vitro. The conditions for RNA IL-2 alpha detection were optimized. The maximum yield of the specific product was obtained at pH 8.5-9.0. The influence of bivalent cations on the efficiency of RT reaction of coupled RT/PCR can be expressed as: Mn2+ > or = Cu2+ > Mg2+ > Cd2+ >> Co2+. The optimal ratio is 1.25-1.88 for Mn2+/dNTPs and 1.88-2.5 for Cu2+/dNTPs and Cd2+/dNTPs. The maximum yield of the RT/PCR product is found at Mg2+/dNTPs = 3.75. When Mn2+ is used instead of Mg2+ in the PCR reaction the efficiency of RT/PCR decreases. The RT/PCR method embracing thermostable Tth DNA-polymerase provides detection of 10(3) copies of RNA IL-2 alpha. An efficient method of the express-diagnostics of MDR-1 gene expression by coupled RT/PCR using Tth DNA polymerase is described. PMID- 7476957 TI - [Purine arabinonucleoside-5'-triphosphates with substituents in the 2'-position as DNA polymerase substrates]. AB - Analogues of araNTPs carrying an azido or aminogroup instead of the 2' hydroxyl exhibited substrate properties towards several mammalian and viral DNA polymerases. At the same time, introduction of a bulky hydrophobic DNP group into the 2' position inactivated the compounds as substrates. HSV-1 and CMV DNA polymerases were an interesting exception: they effectively incorporated the modified nucleotide residues with DNP group into the 3'-termini of the DNA chain. This is a reliable way to distinguish these enzymes from cellular DNA polymerases. PMID- 7476959 TI - [Use of thermostable DNA polymerase from Thermus thermophilus KTP in a combined reverse transcription and amplification reaction for detecting CD4 receptor mRNA]. AB - The RT/PCR method was applied to study a possible use of Tth DNA-polymerase for coupled reaction of reverse transcription and polymerase chain reaction (RT/PCR) on the CD-4 receptor mRNA template in the total cellular RNA. The conditions for detecting the CD-4 receptor mRNA were optimized. The pH-optimum for RT reaction was 8.8. The influence of Mn2+, Cu2+, Co2+, and Cd2+ cations in RT and PCR reaction was investigated. The efficiency of the RT reaction was shown to be the highest in the presence of Mn2+ (optimal concentration 1 mM). At Mn2+ concentration > or = 3 mM complete inhibition of RT/PCR was observed. The Tth DNA polymerase in RT/PCR was shown to be more effective than Taq DNA polymerase. The Tth DNA polymerase allows observation of the specific product in the gel containing ethidium bromide using 20 ng of the total RNA. High sensitivity and specificity of RT/PCR performed with the Tth DNA polymerase allow its wide application in the detection, quantitative analysis and cloning of cellular and viral RNAs. PMID- 7476960 TI - [Effect of an inducer on orientation of the DNA-binding domain of the Lac repressor]. AB - We have identified the lac repressor amino acids which form cross-links with partially depurinated DNA and hence are proximate to it. Only lysine-33 is cross linkable in specific complex with lac operator. At the presence of inducer molecules lysine-33 remains cross-linkable. However, the N-terminal alpha aminogroup was additionally cross-linked with the boundaries of lac operator. We conclude that orientation of the DNA-binding domain of the repressor is reversed by inducer binding. PMID- 7476961 TI - The cyclic adenosine 3',5'-monophosphate-responsive factor CREB is constitutively activated in human somatotroph adenomas. AB - Oncogenic gsp proteins appear to stimulate the transformation of pituitary somatotrophs by inducing the constitutive activation of adenyl cyclase. Previous work implicating the cAMP-responsive transcription factor CREB as a biochemical intermediate in the proliferative response to cAMP led us to examine whether CREB activity was correspondingly elevated in human somatotroph adenomas. In a series of 15 human GH-secreting tumors, we found that each of these contained elevated levels of Ser133-phosphorylated and, hence, activated CREB compared with nonfunctioning pituitary tumors. Four of the GH-secreting adenomas contained an oncogenic gsp gene by polymerase chain reaction analysis, and two additional adenomas expressed wild-type G alpha s protein at 5- to 10-fold higher levels than nonfunctioning pituitary tumors. As both oncogenic gsp and overexpressed G alpha s proteins can induce Ser133 phosphorylation and cAMP-responsive gene expression in transfected GC somatotroph cells, our studies indicate that these proteins may promote somatotroph transformation in part by inducing the transcription of specific CREB-dependent target genes. PMID- 7476962 TI - Cell type-specific regulation of transcription by cyclic adenosine 3,'5' monophosphate-responsive elements within the calcitonin promoter. AB - A cAMP-induced enhancer was previously mapped to nucleotides -255 to -85 of the calcitonin (CT) gene 5'-flanking DNA. To determine the functional cis-acting elements within this region, we transfected medullary thyroid carcinoma (MTC) cells with CT 5'-flanking DNA/GH fusion genes containing potential cAMP responsive elements and assessed their transcriptional activities with and without cAMP. In CT-expressing MTC cells (the TT line), we identified by deletions and point mutations three transcriptionally active motifs: a cAMP responsive element (CRE), TGACGTCA, at -253 to -246, and a hybrid site containing a CRE-like element (CREL; TGACCTCA, -169 to -162) adjacent to an equally transcriptionally active octamer (O) sequence (ATG-CAAAT, -161 to -154). These three motifs acted synergistically and their transcriptional activity was completely dependent on cAMP. In HeLa cells their synergistic activity was more constitutive than cAMP induced, whereas in CT-negative MTC cells (the RO-D81-1 line) these motifs were inactive. Gel mobility shift assays with antibodies against CRE-binding protein (CREB) and activating transcription factor 1 (ATF-1) showed that both CREB and ATF-1 interacted with the CRE in MTC cells, whereas in HeLa cells only ATF-1 bound to the CRE. Specific binding to the CREL/O motif was detectable in extracts from tumors induced by injection of TT cells but not in extracts from any of the three cultured cell lines. We conclude that cAMP-induced transcription of the CT gene is modulated in a cell-specific manner by the CRE and the CREL/O elements. PMID- 7476963 TI - Signaling cross-talk between peroxisome proliferator-activated receptor/retinoid X receptor and estrogen receptor through estrogen response elements. AB - Peroxisome proliferator-activated receptors (PPARs) and retinoid X receptors (RXRs) are nuclear hormone receptors that are activated by fatty acids and 9-cis retinoic acid, respectively. PPARs and RXRs form heterodimers that activate transcription by binding to PPAR response elements (PPREs) in the promoter of target genes. The PPREs described thus far consist of a direct tandem repeat of the AGGTCA core element with one intervening nucleotide. We show here that the vitellogenin A2 estrogen response element (ERE) can also function as a PPRE and is bound by a PPAR/RXR heterodimer. Although this heterodimer can bind to several other ERE-related palindromic response elements containing AGGTCA half-sites, only the ERE is able to confer transactivation of test reporter plasmids, when the ERE is placed either close to or at a distance from the transcription initiation site. Examination of natural ERE-containing promoters, including the pS2, very-low-density apolipoprotein II and vitellogenin A2 genes, revealed considerable differences in the binding of PPAR/RXR heterodimers to these EREs. In their natural promoter context, these EREs did not allow transcriptional activation by PPARs/RXRs. Analysis of this lack of stimulation of the vitellogenin A2 promoter demonstrated that PPARs/RXRs bind to the ERE but cannot transactivate due to a nonpermissive promoter structure. As a consequence, PPARs/RXRs inhibit transactivation by the estrogen receptor through competition for ERE binding. This is the first example of signaling cross-talk between PPAR/RXR and estrogen receptor. PMID- 7476964 TI - Estrogen enhances alpha v beta 3 integrin expression by avian osteoclast precursors via stabilization of beta 3 integrin mRNA. AB - Although bone resorption is accelerated with menopause, the means by which diminished circulating 17 beta-estradiol (E2) promotes osteoclastic activity are unknown. We hypothesized that since the integrin alpha v beta 3 is essential to the resorptive process, reduced E2 levels may increase the integrin's expression by osteoclast precursors. Thus, avian osteoclast precursors (known to contain E2 receptors) were exposed +/- E2, surface iodinated, and lysed. The lysate was immunoprecipitated with an antibody recognizing the intact alpha v beta 3 heterodimer. We find E2 alone fails to impact on alpha v beta 3 expression. Most importantly, however, picomolar (i.e. post-menopausal), but not nanomolar (i.e. premenopausal) concentrations of E2, when added in conjunction with 1,25 dihydroxyvitamin D3 [1,25-(OH)2D3], enhance alpha v beta 3 expression on the plasma membrane of avian osteoclast precursors relative to 1,25(OH)2D3 alone. Induction of alpha v beta 3 by picomolar levels of E2 is dose-dependent, maximizing at 10(-11)-10(-12) M, wherein the sex steroid enhances 1,25-(OH)2-D3 stimulated integrin expression approximately 2.5-fold. Northern analysis reveals that beta 3 mRNA levels parallel those of alpha v beta 3. E2 (10(-12) M) increases expression of beta 3 mRNA induced by a range of 1,25-(OH)2D3 concentrations extending from 10(-10) m-10(-8) M. The E2 + 1,25-(OH)2D3 additive effect on beta 3 mRNA appears as early as 1 day of treatment and progresses for at least 3 days. Consistent with evidence that the beta 3 subunit regulates heterodimer expression, the sex steroid does not impact alpha v mRNA.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7476965 TI - The carboxy-terminal F domain of the human estrogen receptor: role in the transcriptional activity of the receptor and the effectiveness of antiestrogens as estrogen antagonists. AB - Of the steroid hormone receptor family members, the estrogen receptor (ER) is notable in containing a sizable (42-amino acid) C-terminal region, denoted domain F. This F region differs from its adjacent hormone-binding domain, domain E, in that it is not well conserved among different vertebrate ER species, and its role in the biological activity of the ER is not well defined. We report an important role for the F domain of the ER in modulating the magnitude of gene transcription by estrogen and antiestrogen, and in determining the effectiveness of antiestrogens in suppressing estrogen-stimulated gene transcription. Using transient transfections, we have examined, in several cell types, the transcriptional activity of the full-length wild type human ER and ER lacking the carboxy-terminal F domain (delta F ER, containing amino acids 1-554) or ER altered in the F domain by point mutations. In some cells, namely Chinese hamster ovary (CHO) cells and MDA-MB-231 human breast cancer cells expressing wild type ER or delta F ER, estradiol (E2) stimulates equally transcription of several estrogen-responsive promoter-reporter gene constructs [estrogen ca-18119 element, (ERE)2-TATA-CAT, (ERE)2-pS2-CAT, (ERE)2-progesterone receptor(distal)-CAT]; however, the antiestrogens trans-hydroxytamoxifen and ICI 164,384, which stimulate transcription of some of these reporter constructs with the wild type ER, were unable to stimulate transcription with delta F ER. In addition, these antiestrogens were more effective antagonists of E2-stimulated transcription by delta F ER than by wild type ER. By contrast, in HeLa human cervical cancer cells and 3T3 mouse fibroblast cells, the delta F ER exposed to E2 is much less effective than wild type ER in stimulating transcription, and antiestrogens were less potent in suppressing E2-stimulated transcription by the delta F ER. These differences in response of the delta F and wild type ER to estrogen or antiestrogen do not appear to be due to a change in receptor expression level, binding affinity for ligands, or binding to estrogen response element DNA. Our data support the supposition that the conformation of the receptor-ligand complex is different with estrogen vs. antiestrogen and with wild type vs. delta F ER, such that its potential for interaction with protein cofactors or transcription factors is different and is markedly influenced by cell context. Thus, the F domain of the ER has a specific modulatory function that affects the agonist/antagonist effectiveness of antiestrogens and the transcriptional activity of the liganded ER in cells. PMID- 7476966 TI - A somatic cell genetic method for identification of untargeted mutations in the glucocorticoid receptor that cause hormone binding deficiencies. AB - Mouse T lymphoma cell line W7MG1, which is killed by physiological concentrations of glucocorticoid agonists, was used as a convenient genetic system for isolating sublines containing mutant glucocorticoid receptors (GR) with hormone-binding deficiencies. Partially hormone-resistant cell clones were derived from chemically mutagenized cell populations by selecting for growth in moderate concentrations of dexamethasone (Dex) and then screening for failure to grow in high Dex concentrations. Such clones are likely to have mutant GR. In GR cDNA clones from the partially resistant cell sublines, three different functionally significant mutations in the hormone-binding domain were identified: Leu-569 changed to Phe (L569F), Leu-670 to Phe (L670F), and Met-672 to Ile (M672I). Dose response analyses with Dex and affinity labeling studies with dexamethasone 21 mesylate in transiently transfected cells indicated that all three mutant GR species had hormone-binding deficiencies. However, at saturating Dex concentrations the mutant and wild type GR activated a hormone-inducible reporter gene to the same extent; thus, these three mutations did not affect the ability of GR to activate transcription of the reporter gene after hormone was bound. In dose-response curves conducted with several glucocorticoid agonists, mutations L670F and M672I caused no change in ligand-binding specificities, while mutation L569F caused a modest change in specificity. Quantitative hormone-binding studies conducted with mutant GR synthesized in cell-free reactions showed that mutant GR species L569F and M672I had reduced Dex-binding affinities both at 0 C and at 22 C in the presence of molybdate. In contrast, for the L670F mutant, which exhibited the severest deficiency in vivo, the hormone-binding deficiency in the cell-free system was evident only at 26-37 C and primarily in the absence of molybdate. We propose that the L670F GR is an activation-labile type of mutant, which binds hormone normally in the presence of heat shock protein 90 but loses hormone rapidly after dissociation from heat shock protein 90. These three mutations define two new subregions of the GR polypeptide that are important for hormone binding. PMID- 7476967 TI - Interaction of the progesterone receptor with binding proteins for FK506 and cyclosporin A. AB - T47D human breast carcinoma cells and the chicken oviduct were used to study the structure of the nonactivated progesterone receptor (PR) complex. Immunoprecipitation of PR (B form) from cytosol extracts was performed using monoclonal antibody PR6, a cross-reactive antibody prepared to chicken PR. Analysis of the PR complex by sodium dodecyl sulfate gels and Western immuno blotting revealed the presence of several specific copurifying proteins. Consistent with previous reports, the two heat shock proteins, hsp90 and hsp70, were shown to be present. A third 59-kilodalton (kDa) protein observed previously was confirmed to be p59 (also called hsp56 or FKBP52), which has been shown to bind the immunosuppressant drug FK506. Two additional PR-associated proteins were observed that had not been previously recognized with human PR. These have molecular masses of 54-kDa and 23-kDa and have been shown by Western blotting to be related to the proteins p54 and p23 that are associated with chicken PR. P23 is a novel protein of unknown function and p54 or FKBP54 has been recently shown to be another FK506-binding protein related to p59. Finally, the cyclosporin A binding protein, CyP-40, could be detected in isolated chicken PR complexes and in PR complexes that were reconstituted in vitro, but this protein was not detected in human PR complexes, which are less stable than chicken PR complexes in cytosol extracts. The functional significance of FK506 and cyclosporin A binding proteins to hormone action was tested using a T47D cell line that contained a progestin reporter gene, MMTV-CAT. Treatment with cyclosporin A had no effect on the basal level of CAT expression, but it caused a dramatic increase in the sensitivity and magnitude of the response to the synthetic progestin, R5020. The enhanced response elicited by drug treatment was blocked by the antiprogestin RU486 indicating that this effect was receptor-mediated. While cyclosporin A enhanced progestin action in T47D cells, it inhibited a PR/reporter gene system in L cells. The drugs FK506 and rapamycin had no effect on progestin action in T47D cells, but they stimulated glucocorticoid action in T47D cells. Thus, the effects of these immunosuppressant drugs vary with the cell type and hormonal system that is tested. Whether these drug effects relate directly to the immunophilins bound in receptor complexes remains unknown. PMID- 7476968 TI - Regulation of gonadotropin-releasing hormone transcription by protein kinase C is mediated by evolutionarily conserved promoter-proximal elements. AB - We previously demonstrated that down-regulation of protein kinase C (PKC) by prolonged 12-O-tetradecanoylphorbol-13-acetate (TPA) treatment leads to the specific repression of GnRH transcription in GT1-7 hypothalamic neurons. Here we have investigated the regulatory sequences and cognate DNA-binding proteins that mediate this transcriptional response. The promoter-proximal section of the GnRH gene contains an evolutionarily conserved sequence that is bound along its entire length by GT1-7 nuclear proteins in DNase I protection assays. Two distinct regions within this sequence are required for PKC regulation of the GnRH gene, as excision of either region results in loss of TPA repression of transcription. Excision of either of these regions also decreases basal transcription, demonstrating their role in GnRH promoter function. One region encompasses three AT-rich protein-binding sites; the other is an extended region of continuous DNase I protection, 50 nucleotides in length, that contains consensus recognition motifs for the CCAAT/EBP and helix-loop-helix families of transcription factors. Mobility shift analysis of binding to the latter region reveals that TPA treatment of GT1-7 neurons induces the formation of a specific DNA-protein complex with kinetics of appearance consistent with a role in repression of GnRH transcription. Thus, the sequences that mediate PKC regulation of GnRH are proximal to the promoter, evolutionarily conserved, and form TPA-inducible complexes with GT1-7 nuclear proteins. PMID- 7476969 TI - Phosphorylation of the retinoic acid receptor-alpha by protein kinase A. AB - The phosphorylation of retinoic acid receptor-alpha 1 (RAR alpha 1) by PKA was investigated both in vitro and in vivo. We show that bacterially expressed RAR alpha 1 is phosphorylated in vitro by protein kinase A (PKA) at the unique serine residue 369 located in the C-terminal end of the E region. We also show that RAR alpha 1 overexpressed in COS-1 cells is phosphorylated on multiple serine residues and that phosphorylation at serine 369 occurs only when COS-1 cells are cotransfected with PKA or treated with forskolin. RAR alpha 1 mutants were constructed in which serine 369 was replaced by an alanine (S369A) or a glutamic acid (S369E) residue. Comparison of the tryptic phosphopeptide patterns of wild type and mutated RAR alpha 1 overexpressed in COS-1 cells allowed us to confirm that serine 369 is the unique phosphorylation site for PKA in cultured cells. The DNA-binding efficiency of RAR alpha/retinoid X receptor-alpha (RXR alpha) heterodimers was enhanced in vitro by the S369E mutation. However, in transfected RAC65 cells, the same S369E mutation did not affect the ligand-dependent transcriptional activation by RAR alpha 1 of reporter genes containing a retinoic acid (RA)-response element. In contrast, the S369A mutation slightly decreased both DNA binding and the efficiency of PKA to enhance RA-induced transactivation by RAR alpha 1. Finally, we show that endogenous RAR alpha is also phosphorylated in vivo at serine 369 in forskolin-treated F9 cells, supporting the idea that phosphorylation of RARs at this site is involved in the modulation of the RA induced differentiation of F9 cells by (Bu)2cAMP. PMID- 7476970 TI - A naturally occurring short variant of the FTZ-F1-related nuclear orphan receptor xFF1rA and interactions between domains of xFF1rA. AB - The FTZ-F1-related nuclear orphan receptors xFF1rA and B were identified previously in Xenopus laevis by cDNA cloning. In addition to two cDNAs that encode full-length receptor proteins, a third cDNA encodes a form of xFF1rA truncated at the C terminus. Transcripts encoding the short form of the receptor are present at much lower levels than mRNAs encoding the full-length receptors. Significant activation of reporter genes in xFF1rA-transfected HeLa cells requires two or more copies of a FTZ-F1-responsive element (FRE). However, in vitro, recombinant xFF1rA protein binds FRE monomers and dimers with apparently equal affinity. In cotransfection studies, full-length xFF1rA activates transcription, in contrast to xFF1rAshort. In vitro, xFF1rAshort binds to FRE with a lower efficiency than xFF1rA. A partial truncation of the E domain reduces the DNA-binding activity of domain C, suggesting that parts of the E domain might interact with the DNA-binding domain C. In parallel with the loss of DNA-binding efficiency, such truncations lead to loss of transcriptional activation. For transcriptional activation, either the A/B domain or the complete E domain is required, as shown by recombination of different domains of xFF1rA with the DNA binding domain of Gal4. Coexpression of the truncated form xFF1rAshort decreases transcriptional activation by xFF1rA, but not by the active Gal4-xFF1rA fusion protein that contains domain E. This indicates that xFF1rAshort interferes with xFF1A by competition for FRE binding. An excess of xFF1rAshort is required, presumably due to its poor FRE-binding activity. The function of the E domain in regulating DNA-binding and transcriptional activation is discussed. PMID- 7476971 TI - Pituitary-type transcription of the human prolactin gene in the absence of Pit-1. AB - We describe a human (h) PRL-producing cell line, SKUT-1B-20, which we isolated as a subclone of a uterine sarcoma cell line. Although this cell line is of uterine origin, it does not use the decidual-specific upstream promoter of the hPRL gene, but transcribes the hPRL gene from the downstream pituitary-type transcription start site, as determined by Northern blot, reverse transcriptase-polymerase chain reaction and primer extension analyses. This is particularly intriguing because SKUT-1B-20 cells lack the transcription factor Pit-1. No Pit-1 messenger RNA was detectable by reverse transcriptase-polymerase chain reaction, and endogenous Pit-1 target genes (GH, PRL, and Pit-1) were refractory to transfected Pit-1 expression vector, whereas in cotransfection experiments, Pit-1 efficiently activated reporter gene fusion constructs carrying 5'-flanking sequences of the human and rat PRL or the mouse Pit-1 genes. By transfecting reporter genes containing 8.7 kilobases of DNA flanking the hPRL pituitary-specific start site (hPRL-8700/Luc) and deletions thereof, we located a Pit-1-independent cis-active region more than 7 kilobases upstream of the start site. The most distal 1650 or 880 base pairs of the hPRL genomic fragment (which extends to -8784 base pairs), when placed directly upstream of the homologous hPRL or the heterologous thymidine kinase promoters, conferred transcriptional activation to those promoters. SKUT-1B-20 cell-specific activation of hPRL-8700/Luc could not be suppressed by the introduction of an inhibitor of protein kinase A (PKA), PKI. This is the first demonstration of pituitary-type PRL gene transcription independent of Pit-1 and activation of the PKA pathway. The SKUT-1B-20 cell line was then used in reconstitution experiments to delineate the role of Pit-1 in modulating the transcriptional effects of phorbol ester, PKA, and estrogen receptor (ER) on the hPRL gene. The low response of hPRL/luciferase fusion genes to phorbol ester was greatly enhanced by cotransfected Pit-1 and was mediated by the proximal region between -250 and -38. The catalytic subunit of PKA, C beta, was able to elicit a moderate induction of hPRL-8700/Luc even in the absence of Pit-1. A potential estrogen response element has been located in the hPRL gene sequence at a position similar to that of the estrogen response element of the rat PRL gene immediately adjacent to the distal enhancer.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 7476972 TI - Regulation of insulin-like growth factor (IGF)-binding protein-6 and mannose 6 phosphate/IGF-II receptor expression in IGF-IL-overexpressing NIH 3T3 cells. AB - Insulin-like growth factor II (IGF-II)-overexpressing NIH 3T3 cells were used to examine regulation of insulin-like growth factor binding protein (IGFBP) and mannose 6-phosphate (M6P)/IGF-II receptor expression. Ligand blot analysis of conditioned media indicated a predominant IGFBP of 26-28 kilodaltons the abundance of which is 3- to 10-fold higher in media of IGF-II-overexpressing cells. The IGFBP level in control cell medium was increased by incubation in the presence of IGF-II, IGF-I, and mutant IGF-II forms with reduced affinities for IGF-I or M6P/IGF-II receptors. Further proof that IGF-II regulated the IGFBP was obtained by incubation of IGF-II overexpressing cells in the presence of antisense IGF-II oligomers or anti-IGF-II antibodies, which resulted in significant reduction of the IGFBP in conditioned medium. Mouse IGFBP-6 mRNA expression was increased in IGF-II-overexpressing or IGF-II-treated control cells. The IGFBP contained O-linked carbohydrate residues and was recognized by an antiserum to rat IGFBP-6. To determine whether IGFs were influencing proteolytic degradation of IGFBPs, cell-free conditioned media were incubated at 37 C with recombinant human IGFBPs. At neutral pH proteolysis of IGFBP-5 occurred during incubation in conditioned media from control and IGF-II-overexpressing cells. Upon acidification of the medium samples, only the degradation of IGFBP-6 was prevented in IGF-II-overexpressing cell-conditioned medium.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7476973 TI - Rapid activation of insulin-like growth factor binding protein-5 gene transcription during myoblast differentiation. AB - Insulin-like growth factor binding proteins (IGFBPs) comprise a family of secreted proteins that bind insulin-like growth factors-I and -II (IGF-I and -II) with high affinity and potentially modulate their biological effects. We have demonstrated previously that IGFBP-5, the most conserved of the six known IGFBPs, is expressed in muscle cells in the developing embryo and during the terminal differentiation of several myogenic cell lines. In this study we show that an IGF I analog that binds minimally to IGFBPs potently enhances the differentiation of the stringently controlled inducible C2 myoblast (C2l) cell line and identify IGFBP-5 as the sole IGFBP secreted during C2l differentiation. We find that induction of IGFBP-5 mRNA and protein is coincident with the onset of myogenin gene expression and occurs secondary to the rapid activation of IGFBP-5 gene transcription. By transient gene transfer experiments we demonstrate that a 1004 base pair segment of the IGFBP-5 promoter is very active in directing expression of the reporter gene luciferase in C2l myoblasts. A promoter fragment containing only 156 nucleotides of 5'-flanking DNA retained more than 70% of maximal activity and mediated at least part of the differentiation-dependent rise in IGFBP-5 gene transcription. Within this active segment are several potential binding sites for muscle-enriched transcription factors. Our results show that induction of IGFBP-5 expression is an early event in the myogenic differentiation of the C2l cell line and suggest that one function of this IGFBP is to modulate IGF-induced differentiation. C2l cells are thus an excellent in vitro model for elucidating the developmental factors that control IGFBP-5 gene transcription and action in skeletal muscle. PMID- 7476974 TI - Granulosa cell DNA synthesis is strictly correlated with the presence of insulin like growth factor I and absence of c-fos/c-jun expression. AB - Insulin-like growth factor I (IGF-I) mRNA is selectively expressed by a subset of rat ovarian follicles. To elucidate the biological role and regulation of follicular IGF-I, we have analyzed individual follicles with respect to IGF-I production, DNA synthesis, and c-fos and c-jun expression, since AP-1 complex components have been implicated both in regulation of IGF-I gene expression and cell proliferation. IGF-I mRNA localization, bromodeoxyuridine (BRDU) incorporation, and c-fos and c-jun immunoreactivity were compared in serial sections from prepubertal and mature rat ovaries. BRDU incorporation is exclusively concentrated in IGF-I-expressing follicles in both prepubertal and mature rats, with further intrafollicular correlation between local concentrations of BRDU-positive granulosa cells and IGF-I mRNA. Furthermore, there is a striking negative correlation between IGF-I mRNA localization and follicular fos/jun immunoreactivity. Both c-fos and c-jun are readily detected in granulosa cells of luteinized follicles where IGF-I mRNA is never found. Immunoreactive c-jun but not c-fos is detected in granulosa cells of atretic follicles, while neither c-fos nor c-jun is detected in IGF-I mRNA-positive granulosa cells. These data implicate IGF-I in granulosa cell proliferation and the fos/jun heterodimer in granulosa cell luteinization, while expression of c jun in the absence of c-fos may be implicated in programmed granulosa cell death. PMID- 7476975 TI - Targeted ablation of pituitary pre-proopiomelanocortin cells by herpes simplex virus-1 thymidine kinase differentially regulates mRNAs encoding the adrenocorticotropin receptor and aldosterone synthase in the mouse adrenal gland. AB - We have produced and characterized lines of transgenic mice expressing a fusion gene composed of the pituitary expression-specific promoter region of the POMC gene, driving the herpes simplex viral-1 thymidine kinase. Adult mice were treated with the antiherpes agent ganciclovir at 70 mg/kg body weight (ip, twice daily for 10-12 days). Approximately 98% of the pituitary intermediate lobe melanotropes and anterior lobe corticotropes were ablated as determined by immunocytochemistry and RIA specific for the POMC-derived peptides, ACTH, beta endorophin, and alpha-MSH. The number of lactotropes, somatotropes, thyrotropes, and gonadotropes was not altered compared with controls, indicating that in the adult pituitary, POMC products are not required to maintain the distribution of cell types. As expected, plasma corticosterone levels were substantially decreased after POMC cell ablation. In situ hybridization studies showed that the mouse ACTH receptor was expressed uniformly throughout the adrenal cortex, and RNase protection assays revealed that the ACTH receptor mRNA decreased after pituitary POMC cell ablation. Additionally, RNase protection assays showed that pituitary POMC cell ablation resulted in the decrease of adrenal p450c11 beta transcripts while p450c11AS (aldosterone synthase) mRNA levels remained constant. These data demonstrate differential regulation of steroid pathway-specific enzymes by POMC products. Our results also suggest that the thymidine kinase cell obliteration technique may not be dependent on cell division as a prerequisite for cytotoxicity, thus supporting the idea that targeted molecular ablation using cell- and tissue-specific promoter sequences to drive viral thymidine kinase expression can be refined further to study other nonmitotic cells. PMID- 7476976 TI - Androgen receptor-mediated transcriptional regulation in the absence of direct interaction with a specific DNA element. AB - Androgen receptor (AR) brings about a ligand-dependent inhibition of low-affinity neurotrophin receptor (p75) promoter constructs in cultured cells, with the greatest inhibition being achieved with a reporter gene containing 1050 nucleotides (nt) of the promoter. The receptor domain critical for trans repression localizes to the same region (amino acids 147-296) as that mandatory for transactivation. In contrast to trans-activation, AR does not interact directly with specific DNA elements to elicit trans-repression of p75 promoter constructs, although an intact DNA-binding domain of the receptor is required for both actions. In a search for interacting partners, both extensively purified full-length AR and AR-DNA binding domain were found to inhibit c-Jun/AP-1 site interaction without themselves binding to the AP-1 element. Prior binding of c Jun to the AP-1 element protected the complex from the receptor's interference. Repression was not mutual, as c-Jun did not inhibit AR-androgen response element interaction or trans-activation through an androgen response element-containing promoter. The 1050-nt-long p75 promoter sequence does not contain an AP-1 element; an AP-1-like site in the vector backbone mediates the trans-repression by the AR in recipient cells. Intriguingly, an AR form with a large N-terminal deletion (the delta 46-408 mutant) behaved as a transcriptional activator of the p75 promoter through a mechanism that was also independent of specific DNA binding. Collectively, these data indicate that, in a proper context, AR is able to elicit both transrepression and trans-activation without interacting directly with specific DNA elements. Sequences responsible for the down-regulation of p75 mRNA by androgens in vivo are, however, not located in the proximal 1050 nt of the p75 promoter. PMID- 7476977 TI - Identification of a group of Ser-Pro motif hormone-inducible phosphorylation sites in the human progesterone receptor. AB - The human progesterone receptor (PR) is a member of the steroid/thyroid hormone superfamily of nuclear receptors. The receptor is expressed as two forms, PR-B and the shorter PR-A, which lacks the NH2-terminal 164 amino acids of PR-B; whereas PR-B seems to be predominantly a transcriptional activator, PR-A also functions as a repressor. Our previous studies of PR expressed in T47D breast cancer cells have shown that PR is a phosphoprotein whose phosphorylation is enhanced in response to hormone. There is an initial rapid (minutes) increase in phosphorylation followed by a slower, less substantial increase, which results in decreased mobility of the receptor on sodium dodecyl sulfate gels. We now report the identification of three phosphorylation sites, which are predominantly phosphorylated during the later phase of the response to hormone. These sites, Ser102, Ser294, and Ser345, are all found in Ser-Pro consensus sequences. Whereas Ser294 and Ser345 are common to PR-A and PR-B, Ser102 is unique to PR-B. Finally, we demonstrate that phosphorylation of Ser345 is associated with the altered mobility on sodium dodecyl sulfate gels. PMID- 7476978 TI - Estradiol and phorbol ester cause phosphorylation of serine 118 in the human estrogen receptor. AB - Serine 118 is definitively identified as a major site of phosphorylation in the human estrogen receptor expressed in COS-1 cells treated with estradiol or phorbol ester. At least 30% of the estrogen receptor appears to be phosphorylated on serine 118 after treatment with estradiol or phorbol ester. Human estrogen receptor was expressed in COS-1 cells and labeled in vivo with [32P]orthophosphate in the presence of estradiol or phorbol ester. Immunopurified receptor was digested with cyanogen bromide. The most heavily labeled peptide (7 kilodaltons) was identified as amino acids 110-174 by microsequencing. Manual Edman degradation released a major portion of the 32P-label in the peptide at serine 118. A mutant with serine 118 replaced by alanine (S118A) had 80% less 32P label in the 7 kilodalton peptide. Estrogen receptor labeled in vivo with [32P] orthophosphate in the presence of estradiol or phorbol ester migrates electrophoretically as a doublet. The major difference between the bands is phosphorylation of serine 118 in the upshifted band. The mutant S118A does not show an upshifted band. Labeling of the estrogen receptor with [35S]methionine indicates that > or = 30% of the receptor is upshifted and suggests that > or = 30% of the receptor is phosphorylated on serine 118. PMID- 7476979 TI - A naturally occurring estrogen receptor mutation results in increased estrogenicity of a tamoxifen analog. AB - We previously identified a codon 351 (Asp-->Tyr) mutant estrogen receptor (ER) in a tamoxifen-stimulated human breast tumor line. To examine its biological activity, we have constructed cell lines from the ER-negative human breast cancer cell line MDA-MB-231 that stably express either the wild type (S30) or mutant ER (BC-2). ER expression was confirmed by Western blot, ligand-binding studies, and ER-enzyme immunoassay. The growth characteristics of the S30 and BC-2 cell lines were compared when treated with estradiol, fixed-ring 4-hydroxytamoxifen [(fr) 4 OH TAM], or ICI 182,780. (fr) 4-OH TAM is a stable, high affinity tamoxifen analog. Many investigators have recognized that growth of ER-negative cell lines stably transfected with ER is inhibited by estradiol. Similarly, both S30 and BC 2 cell lines are inhibited by estradiol in a concentration-dependent manner. (fr) 4-OH TAM has no effect on S30 proliferation but inhibits the growth of BC-2 cells. The pure antiestrogen ICI 182,780 can block the growth-inhibitory effect of estradiol in both cell lines and the growth-inhibitory effect of (fr) 4-OH TAM in the BC-2 cells. In transient transfection analyses using a luciferase reporter plasmid containing two copies of the Xenopus vitellogenin A2 estrogen response element, estradiol stimulated luciferase transcription through both the wild type and mutant estrogen receptors, while (fr) 4-OH TAM stimulated transcription to a greater extent through the mutant receptor. These results demonstrate that the estrogenicity of (fr) 4-OH TAM is increased by binding to the codon 351 mutant ER, and that ER activation and growth inhibition are associated. PMID- 7476980 TI - Estrogen receptor-induced DNA bending: orientation of the bend and replacement of an estrogen response element with an intrinsic DNA bending sequence. AB - The estrogen receptor (ER) binds to DNA fragments containing estrogen response elements (EREs) and causes them to bend. To characterize this ER-induced DNA bend and determine if it is involved in transcription activation, three different lines of investigation were used. Using MCF-7 human breast cancer cell extracts and circular permutation analysis, it was determined that molybdate-stabilized, unoccupied cytosolic ER was unable to bind to ERE-containing DNA fragments when maintained at 4 C, but that thermal activation enabled the cytosolic receptor to bind and bend ERE-containing DNA fragments to the same extent as ER present in whole cell extracts. DNA phasing analysis was utilized to determine that ER binding induced DNA fragments containing EREs to bend toward the major groove of the DNA helix. The orientation of this bend was the same with thermally activated, unoccupied cytosolic ER and with unoccupied ER, 17 beta-estradiol occupied ER, and 4-hydroxytamoxifen-occupied ER present in whole cell extracts. Using transient cotransfection assays, the ability of an intrinsically bent DNA sequence to replace an ERE was tested. When a single consensus ERE, which is induced to bend 56 degrees on ER binding, was replaced with a 54 degrees intrinsic DNA bending sequence, transcription was effectively activated. Similar levels of transcription were also observed when promoters contained either a 108 degrees intrinsic DNA bending sequence or two consensus EREs. However, the 54 degrees DNA bending sequence and a single ERE were unable to cooperatively activate transcription. Because the magnitude and orientation of ER-induced DNA bends are the same with the unoccupied and occupied receptor, DNA bending alone probably does not function as a transcriptional switch to turn on gene transcription. However, DNA bending may be required to provide the architecture needed for modulation of target genes. PMID- 7476981 TI - In vivo functional analysis of the mouse estrogen receptor gene promoter: a transgenic mouse model to study tissue-specific and developmental regulation of estrogen receptor gene transcription. AB - Understanding the molecular and morphological basis of estrogen responsiveness in the various tissues and organs that make up an adult organism and its onset during ontogenesis requires identification of the genetic controls that determine timed expression of the estrogen receptor (ER) gene in multiple cell types. With this goal in mind, we describe here the results of the functional analysis of the mouse (m) ER gene promoter, carried out in vivo in transgenic mice. The mER gene promoter was cloned and spliced to the coding sequence of the bacterial lacZ gene (fused to the nuclear localization signal of SV40 large T: nls-beta-GAL) and then stably reintegrated into the genome of mice. Analysis of beta-GAL mRNA and protein expression in multiple organs of both female and male transgenic animals was then performed. Results show that the transgenic mER promoter, much like the endogenous one, is active in several organs and tissues of adult female and male mice. The first 0.4 kilobases of 5'-flanking DNA (up to -364) are sufficient to direct widespread expression of the transgene in mouse organs. This indicates that genetic elements functional in various cell types are included in this segment. Furthermore, the first exon and intron of the mER gene are necessary to achieve sexually dimorphic expression of the transgene in neurons located at specific sites within the central nervous system. These mER promoter transgenic mice will be useful in mapping estrogen- responsive cell types under different physiological and pathological conditions in vivo, in defining ontogenesis of estrogen action in the mouse, and in studying the mechanisms that regulate ER gene transcription. PMID- 7476982 TI - Cholesterol side-chain cleavage cytochrome P450 gene expression in the primitive gut of the mouse embryo does not require steroidogenic factor 1. AB - In situ hybridization studies reveal novel sites of expression of cholesterol side-chain cleavage cytochrome P450 (P450scc) during murine embryonic development. In addition to fetal adrenals and testes, P450scc transcripts localize in situ to the primitive gut and to a subset of unidentified cells in the dermal mesenchyme of embryonic skin. In the gut, transcripts are most abundant in luminal epithelia of the hindgut, which will form the colon. P450scc transcript abundance at these novel sites is a fraction of that in fetal adrenals or testes, suggesting a local rather than an endocrine function. Immunocytochemical analyses localize P450scc protein to the fetal hindgut, indicating that the transcripts are translated in vivo. RNA isolated from microdissected embryonic hindgut and skin was reverse transcribed and amplified by polymerase chain reaction. DNA sequence analyses of polymerase chain reaction products confirmed that specific hybridization in situ represents authentic P450scc gene (Cyp11A) transcripts and that 3 beta-hydroxysteroid dehydrogenase/delta 5-->delta 4-isomerase transcripts are also present, demonstrating the potential of these fetal tissues to produce pregnenolone and progesterone. P450scc transcripts are also detectable by in situ hybridization in primitive gut and skin of Fushi tarazu factor 1 null mice, which lack the nuclear receptor steroidogenic factor 1, proving that steroidogenic factor 1 is not required for steroid hydroxylase gene expression at these sites. The capacity for C21 steroid biosynthesis in primitive gut and skin during organogenesis raises the question whether local production of steroid hormones may be required for normal cellular growth and differentiation of these tissues during embryogenesis. PMID- 7476984 TI - Citation for the 1995 Edwin B. Astwood Lecture Award to Kenneth S. Korach. PMID- 7476983 TI - Citation for the 1995 Fred Conrad Koch Award of the Endocrine Society to Jack Gorski. PMID- 7476985 TI - Citation for the 1995 Rhone-Poulenc-Rorer Clinical Investigator Award to Maria I. New. PMID- 7476986 TI - Citation for the 1995 Sidney H. Ingbar Distinguished Service Award of the Endocrine Society to Jacob Robbins and Joseph Edward Rall. PMID- 7476987 TI - Citation for the 1995 Gerald D. Aurbach Lecture Award to Robert Joseph Lefkowitz. PMID- 7476988 TI - Citation for the 1995 Robert H. Williams Distinguished Leadership Award of the Endocrine Society to David N. Orth. PMID- 7476990 TI - Citation for the 1995 Richard E. Weitzman Memorial Award of the Endocrine Society to Mitchell Avery Lazar. PMID- 7476989 TI - Citation for the 1995 Ernst Oppenheimer Memorial Award of the Endocrine Society to Deborah L. Segaloff. PMID- 7476991 TI - Modulation of a Ca(2+)-activated K+ channel by angiotensin II in rat adrenal glomerulosa cells: involvement of a G protein. AB - In the present study, we demonstrate the presence of Ca(2+)-activated K+ channels in rat glomerulosa cells. We find that angiotensin II (Ang II) inhibits this charybdotoxin-sensitive current. The effect of Ang II was dose-dependent with an inhibition constant (Ki) of 0.98 nM and a maximal effect observed at 200 nM. Time course of the blockage was as rapid as the one induced by charybdotoxin. This effect is mediated by the AT1 receptor subtype of Ang II, since it is blocked by DUP 753 but is unaffected by CGP 42112. Activation of protein kinase C by phorbol dibutyrate (1 microM) or dialysis of the cell with inositol 1,4,5-triphosphate (20 microM) were ineffective in blocking the current. However, experiments done with GDP beta S and GTP gamma S indicated that a G protein was involved. The inhibitory effect of Ang II was not pertussis toxin-sensitive, which excludes Gi protein, but was abrogated if an antibody raised against the alpha-subunit of the Gq/11 protein was present in the patch pipette medium. Further analysis showed that the Ca(2+)-activated K+ channel was able to modulate the membrane potential according to the level of intracellular calcium concentration ([Ca2+]i). Whereas a thapsigargin-induced increase in [Ca2+]i hyperpolarized the membrane, this effect was not observed when Ang II was used to increase [Ca2+]i because of the blockage of the Ca(2+)-activated K+ current. The blockage of Ca(2+)-activated K+ current by Ang II would result in a synergistic effect on the Ang II-induced depolarization, thus favoring Ca2+ influx, an event essential to secretion. PMID- 7476992 TI - Role of the carboxy-terminal residues of the alpha-subunit in the expression and bioactivity of human thyroid-stimulating hormone. AB - The glycoprotein hormones TSH, CG, LH, and FSH are heterodimers consisting of a hormone-specific beta-subunit and a common alpha-subunit. The aim of the present study was to investigate the role of the carboxy terminus of the common alpha subunit (amino acids Tyr89-His90-Lys91-Ser92), which has been shown to be important for human (h) CG and hFSH, for the activity of hTSH. Successive truncations of the alpha-carboxy terminus by site-directed mutagenesis revealed a stepwise reduction of bioactivity occurring at residues alpha Ser92 and alpha His90 to 64% and 13%, respectively. This contrasts with previous findings for hCG and hFSH, where loss of bioactivity occurred in a single step with the deletion of alpha Lys91 but alpha Ser92 was not important. The decreased bioactivities of the hTSH alpha-truncation mutants were reflected by concomitant reductions of cAMP production, thyroid hormone synthesis and cell growth and were accompanied by a loss of receptor binding. Substitution of residues alpha Lys91 or alpha His90 with either a hydrophobic or a bulkier residues resulted in a reduction of receptor binding and signal transduction, indicating that the alpha-carboxy terminus of hTSH may interact with the TSH receptor in a tight contact area. Conversely, substitution of alpha His90 with smaller residues enhanced bioactivity. In addition, the integrity of the alpha-carboxy terminus was essential for hTSH expression. Thus, we showed common and different roles of the alpha-carboxy-terminal residues for the glycoprotein hormones. The unique role of alpha Ser92 in hTSH activity explains the evolutionary constraint to preserve the alpha-carboxy-terminal Ser92 in all glycoprotein hormones. PMID- 7476993 TI - Functionally different isoforms of the human calcitonin receptor result from alternative splicing of the gene transcript. AB - Two subtypes of the human calcitonin receptor (hCTR) have been described which differ from one another by the presence or absence of a 16-amino acid insert in the first intracellular loop. Both isoforms were stably expressed in baby hamster kidney cells to compare their ligand binding and second messenger coupling. The binding affinity and the on/off rate of binding for salmon CT were identical for the two receptor isoforms. However, the presence of the insert significantly reduced the ability of the receptor to couple to both adenylate cyclase and phospholipase C. Stimulation of a transient calcium response was only observed with the insert-negative receptor. Similarly, the ED50 for the cAMP response is 100-fold higher for the insert-positive form compared with the insert-negative form of the receptor. However, the maximal cAMP response was equivalent for both receptor isoforms. The rate of internalization of the insert-positive form of the receptor is significantly impaired relative to the insert-negative receptor, which suggests that this process may be dependent on the stimulation of a second messenger pathway. Cloning and characterization of the relevant portion of the hCTR gene revealed that these isoforms are generated by alternative splicing. We also discovered a third isoform of the hCTR, which can be generated by alternative splicing at the same position. The presence of a stop codon in this newly described alternative exon would lead to premature termination of the receptor at the C-terminal end of the first transmembrane domain. PMID- 7476994 TI - Structural and functional differences in the dio1 gene in mice with inherited type 1 deiodinase deficiency. AB - The type 1 deiodinase (D1) provides the major portion of the circulating T3 in vertebrates. In C3H and certain other inbred mice, liver and kidney D1 activity is 5- to 10-fold lower than in the common phenotype, C57. The lower D1 levels are paralleled by a decreased normal-sized dio1 mRNA and hyperthyroxinemia. Low activity cosegregates with a restriction fragment length variant (RFLV) in both inbred and recombinant strains, indicating it is due to differences in the dio1 gene. The exonic structure and the deduced amino acid sequences are identical for both strains and highly homologous to that of the rat. The RFLV is due to an approximately 150-base pair expansion of repetitive sequences in the second intron of the C3H gene, but this segment does not differentially affect the transient expression of a human GH gene. The promoter and 5'-flanking regions of the C3H and C57 dio1 genes are very similar and are GC rich without TATA or CCAAT boxes. However, functional assays of 1.5-kilobase 5'-flanking dio1-CAT constructs showed 2- to 3-fold higher activity of the C57-CAT constructs. Deletion mutants showed that sequences between -705 and -162 were the cause of this. In this region, the only major difference between the two genes is a 21-base pair insert containing five CTG repeats in the C3H promoter. This difference also cosegregates with low D1 activity and the intron RFLV in four other mouse strains. The correlation of the CTG repeat insert with both in vitro and in vivo expression and the absence of other significant sequence differences in the 5' flanking region argue that this is the major explanation for the impaired expression of the dio1 gene and the resulting hyperthyroxinemia of the C3H mouse. PMID- 7476995 TI - Synergistic activation of neurotensin/neuromedin N gene expression by c-Jun and glucocorticoids: novel effects of Fos family proteins. AB - The cis-regulatory region of the neurotensin/neuromedin N (NT/N) gene integrates diverse environmental signals in the neuroendocrine PC12 cell line, resulting in remarkable synergistic regulation. An AP-1 site appears to play a pivotal role in cooperative NT/N gene activation, as mutations in this site decrease responses to all inducer combinations by at least an order of magnitude. Here we report that c Jun acts synergistically with glucocorticoids to activate the NT/N promoter, and that Fos family proteins have novel regulatory effects on this interaction. Cotransfection of individual pCMV-AP-1 expression plasmids revealed that c-Jun most potently activates the NT/N promoter and that costimulation with dexamethasone results in a further 6- to 12-fold increase in expression. Unlike its general inhibitory effects on glucocorticoid regulation in other systems, c Fos potentiated activation by glucocorticoids when coexpressed with c-Jun, and Fos B had a similar, but more limited, positive effect. In contrast, Fra-1 reversed the direction of glucocorticoid regulation, and Fra-2 abolished synergism. AP-1, cAMP response element, and glucocorticoid response element motifs are required for full cooperative activation by either c-Jun or c-Jun/c Fos and glucocorticoids. These results indicate that NT/N promoter activation involves synergistic interactions between specific AP-1 complexes and ligand activated glucocorticoid receptor, and similar mechanisms may regulate NT/N gene expression in central neurons. PMID- 7476996 TI - Lactate dehydrogenase A subunit messenger RNA stability is synergistically regulated via the protein kinase A and C signal transduction pathways. AB - We have identified and studied a posttranscriptional mechanism of lactate dehydrogenase A (LDH) subunit gene expression at the level of mRNA stability. Using the well differentiated rat C6 glioma cell line as a model system, the effects of activators of the protein kinase A and C pathways on the half-life of LDH A mRNA were measured by two independent methods: 1) by the RNA synthesis inhibitor-chase method using actinomycin D, and 2) by analysis of decay of LDH A [3H]mRNA in [3H]uridine-labeled cells. By each method, the half-life of relatively short-lived LDH A mRNA was increased 5- to 7-fold in 8- (4-chloro phenylthio) cAMP or forskolin-treated and about 3-fold in 12-0 tetradecanoylphorbol-13- acetate (TPA) or dioctanoylglycerol-treated cells. Forskolin acted synergistically with TPA to prolong LDH A mRNA half-life from 55 min to more than 20 h. The relatively rapid basal decay rate of LDH A mRNA was also considerably slowed in the presence of the protein phosphatase inhibitor okadaic acid, suggesting a functional role for protein phosphorylation in the stabilization process. In glioma cells stably transformed with a protein kinase A catalytic subunit expression vector, overexpression of the catalytic subunit stabilized LDH mRNA to the degree seen in forskolin-treated cells. In cells transfected with a protein kinase A inhibitor-expression vector, cAMP-mediated stabilization of LDH A mRNA half-life was prevented. Furthermore, both staurosporin and 3- [1-(3-dimethylaminopropyl)-indol-3-yl]-3-(indol- 3-yl)- maleimide, inhibitors of protein kinase C, prevented the TPA-induced stabilization of LDH A mRNA. We conclude from the experimental data that the protein kinase A and C signal pathways play an active functional role in regulating LDH A mRNA stability and act cooperatively to achieve LDH A mRNA stability regulation. PMID- 7476997 TI - Modelling antibody-antigen interactions: ferritin as a case study. AB - In this work, we propose a model for the structure of the antigen-antibody complex formed by human H-ferritin and an antibody that specifically recognizes it. We cloned and sequenced the antibody gene, predicted the antibody three dimensional structure, and reconstructed the H-ferritin-antibody complex using an automated docking procedure previously validated on known complexes. This procedure allowed us to identify one putative complex which we carefully analysed, in order both to evaluate its likelihood, in light of a set of experimental results described in the literature, and to predict precisely which are the sites of interaction between the two molecules. Our model is compatible with the experimentally determined characteristics of the complex. Some of the residues that form the predicted antigenic site of ferritin can be found in the amino acid sequence of peptides selected from a random peptide library because of their affinity for the ferritin monoclonal antibody. Furthermore, the structural difference between the antigenic site in human H-ferritin and the corresponding region in other species permits us to rationalize the inability of the antibody to recognize human L-ferritin and rat, chicken and mouse H-ferritin. Through the analysis of our model complex, we identify a number of other residues putatively involved in the interaction. This multidisciplinary approach shows that synergy between computational and experimental methods may bring further insight into the understanding of antibody-antigen recognition rules. PMID- 7476998 TI - Glycosylation of the variable region of immunoglobulin G--site specific maturation of the sugar chains. AB - The structure of the N-linked sugar chains attached to three IgG antibodies, identical in amino acid sequence except for the changes required to introduce the carbohydrate addition sites, has been determined. All three antibodies are specific for dextran but differ in their ability to bind antigen. The heavy chains with a murine variable region (V region) attached to the human gamma 4 constant region were expressed in a murine hybridoma synthesizing the specific light chain. In addition to the glycosylation site in the Fc portion, each antibody has a different glycosylation site in the second complementarity determining region (CDR2) of the heavy chain (Asn54, Asn58, or Asn60). The sugar chains were released from purified Fab and Fc fragments by hydrazinolysis and converted to radioactive oligosaccharides by reduction with sodium borotritide. The structures of these radioactive oligosaccharides were determined by a combination of sequential exoglycosidase digestion and Bio-Gel P-4 and lectin column chromatography. For all three antibodies, the carbohydrate attached to the Fc portion was a mixture of complex-type biantennary sugar chains. The variable region carbohydrate structures attached at Asn54 and Asn58 were also complex-type but more highly sialylated than were the Fc-associated sugars. Moreover, unlike the Fc-associated sugars, a significant population of Fab-associated sugars contained a Gal alpha 1-->3 residue as a non-reducing terminus. In contrast, the carbohydrate attached at Asn60 was a high mannose structure. These results demonstrate that slight changes in the position of carbohydrate attachment within CDR2 of the variable region of the heavy chain can substantially alter carbohydrate processing and that complex-type carbohydrates contained within the same polypeptide chain can have different structures. PMID- 7476999 TI - How do multichain immune recognition receptors signal? A structural hypothesis. AB - Several cell surface receptors involved in cellular activation by antigen, such as the B-cell and T-cell antigen receptors, and receptors for IgE and IgG (Fc epsilon RI and Fc gamma RIII) show substantial similarities in structure and signaling pathways. An essential step in the activation of immune cells through these receptors is the phosphorylation of specific tyrosine residues within certain consensus sequences found in the cytoplasmic tails of different chains belonging to each of these receptors. The mechanism by which aggregation of the receptors triggers these phosphorylation is still unknown. In this paper, a mechanistic model for this key event is proposed. This model assumes that the kinase(s) responsible for catalysing these phosphorylations do exist associated with the receptors, but for steric reasons they cannot phosphorylate tyrosine residues on chains of the same receptor complex. Upon aggregation, these kinases phosphorylate the tyrosines of a distinct receptor complex (cross phosphorylation), thus starting the signaling cascade. PMID- 7477000 TI - Activation-induced apoptosis in Jurkat cells through a myc-independent mechanism. AB - Apoptosis can be induced in the human leukemic T-cell line Jurkat when these cells are stimulated by several known T-cell activators. Among the stimuli that activate Jurkat cells, calcium ionophore A23187, phytohemagglutinin and cross linked monoclonal antibody anti-CD3 are the best inducers of apoptosis. The activator of protein kinase C phorbol-12,13-dibutyrate causes a rapid but transient stimulation of DNA fragmentation and cell death, and soluble monoclonal antibody anti-CD3 has no effect. Furthermore, apoptosis is delayed when Jurkat cells are stimulated simultaneously with calcium ionophore and phorbol ester. We have analysed the mechanism underlying the induction of apoptosis by these agents and found that down-regulation of c-myc expression by pretreatment with the DNA binding antibiotic mithramycin, an inhibitor of c-myc transcription, does not prevent the activation of the apoptotic process. In the light of these results we can conclude that in these leukemic T-cells, activation of apoptosis may occur by a mechanism independent of c-myc expression. PMID- 7477001 TI - Activation of human complement by totally human monoclonal antibodies. AB - A uniquely developed series of totally human monoclonal antibodies (mAbs) were examined for their complement fixing properties in comparison to human myeloma preparations and to commercially available human polyclonal immunoglobulins. C3b and C4b deposition was measured using a kinetic ELISA technique. When the IgG myeloma proteins were tested for classical pathway activation, our findings were similar to those previously described, where IgG1 and IgG3 were more potent activators of the classical pathway than IgG2 and IgG4. However, those same studies determined that IgG2 was the best activator of the alternative pathway followed by IgG1 and IgG3 while IgG4 does not activate complement via either pathway. In our studies of alternative pathway activation, the IgG2 myeloma exhibited strong activation of the alternative pathway, but, at levels lower than the other three IgG subtypes. Using this test system, we examined the complement activating potential of four totally human mAbs that were constructed from the peripheral blood lymphocytes of a colon carcinoma patient in long term remission. We found that our uniquely constructed totally human IgG2 mAbs (A3, E1, F6 and F8) were able to activate complement by both the classical and alternative pathways to varying degrees. In addition, we found that the complement activating ability of the human mAbs was greater than that of the human IgG2 myeloma immunoglobulins or normal human IgG2 preparations. This study represents the first report of complement activation by totally human mAbs and confirms more recent findings which indicate that levels of complement activation by human IgG immunoglobulins cannot be predicted based solely on their subclass identity. PMID- 7477002 TI - Involvement of nitric oxide in IFN-gamma-mediated reduction of microvessel smooth muscle cell proliferation. AB - Previous studies in our laboratory demonstrated that murine cerebral microvessel smooth muscle cells (SMC) activate syngeneic CD4+ T-cells in vitro. These T cells, or their culture supernatants, in turn, strongly inhibit proliferation of the SMC. The present study focuses on IFN-gamma as a mediator of inhibition of SMC proliferation, and addresses the molecular mechanism of this inhibition. IFN gamma profoundly reduced the proliferation of murine brain microvessel smooth muscle cells in vitro. Three lines of evidence indicate that nitric oxide contributed to this effect: (1) IFN-gamma-mediated inhibition of proliferation correlated with the quantity of nitrite, a stable breakdown product of nitric oxide, in culture supernatants; (2) the addition of N(g)- monomethyl-l-arginine, and inhibitor of nitric oxide synthesis, restored proliferation to control or near control levels; and (3) the addition of hemoglobin, which has a high affinity for, and thus sequesters nitric oxide, also resulted in significant restoration of the proliferative response. However, the nitric oxide donating chemical sodium nitro-prusside, at concentrations up to 100 microM, had no direct cytostatic effect. These results suggest that nitric oxide is a necessary but insufficient component in IFN-gamma-mediated inhibition of microvessel smooth muscle cell proliferation. TNF-alpha also stimulated nitric oxide production by the smooth muscle cells, but was not as potent as IFN-gamma at inhibiting proliferation. Knowledge of the physiological effects of lymphokines on cells of the brain microvasculature will contribute towards a better understanding of inflammatory processes in diseases such as multiple sclerosis and infectious encephalitis. PMID- 7477003 TI - Clip binds to HLA class II using methionine-based, allele-dependent motifs as well as allele-independent supermotifs. AB - The invariant chain (Ii) region that interacts with class II and inhibits premature peptide binding has been mapped to amino acids 82-107, known as CLIP. It is unclear whether CLIP binds directly to the class II peptide binding groove and thus competitively blocks binding of other peptides, or whether it binds to conserved class II sites and indirectly inhibits peptide binding by inducing conformational changes in class II. Here we show evidence that strongly suggests that CLIP binds within the peptide binding groove, as CLIP binds to various HLA DR alleles using allele-dependent as well as allele-independent, methionine-based binding motifs. First, a core sequence of 12 amino acids was identified within CLIP which is required for optimal binding to DR1, DR2, DR3(17) and DR7. This sequence is composed of CLIP p88-99 (SKMRMATPLLMQ). By substitution analysis, all three methionine residues appeared to control CLIP binding to HLA-DR. However, whereas M90 controlled binding to all four alleles, M92 and M98 were of different importance for the various alleles: M92 is involved in CLIP binding to DR1 and DR3(17) but not to DR2 or DR7, and M98 controls CLIP binding to DR2, DR3(17) and DR7 but not DR1. Also, CLIP competes with known immunogenic peptides for class II binding in a manner indistinguishable from regular, class II binding competitor peptides. Finally, the dissociation rates of CLIP-class II complexed are similar to those of antigenic peptide-class II complexes. Thus, CLIP most likely binds to the class II peptide binding groove, since most allelic class II differences are clustered here. CLIP uses unconventional methionine anchor residues representing an allele-independent supermotif (M90) as well as allele-dependent motifs (M92 and M98). PMID- 7477004 TI - Role of conserved amino acid residues in the complementarity determining regions on hapten-antibody interaction of anti-(4-hydroxy-3-nitrophenyl) acetyl antibodies. AB - Monoclonal antibodies (mAbs) specific to (4-hydroxy-3-nitrophenyl)acetyl (NP) were prepared at various times after immunization and the amino acid sequences of VH and V lambda 1 in these mAbs were deduced from cDNA nucleotide sequences. Replacements due to somatic mutation were not found in day 7 mAbs but were found in those of days 14, 84 and 294. The affinity of day 7 mAbs to NP-glycine(NP-Gly) was in the order of 10(4) M-1 and it increased about 8000-fold with time after immunization. The extrinsic circular dichroism (CD) spectrum of the NP-epsilon aminocaproic acid (NP-Cap)/Ab complex was unique for each mAb, although the spectra were grouped into two types, which tended to shift from one type to another with time, suggesting a variation in the micro-environments around NP-Cap in the combining sites. All these data indicate that the structure of the combining site was altered by somatic mutation; however, the fine-specificity measured by cross-reactivity with hapten analogues did not change significantly with time. We examined the amino acid residues in CDRs responsible for recognition of NP-haptens by comparing the amino acid sequences of anti-NP mAbs. Analyses revealed the presence of several conserved amino acid residues in CDRs of VH and V lambda 1, such as Tyr-32H, and Tyr-60H, in addition to a core segment involving Arg-50H.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7477005 TI - A synthetic standard DNA construct for use in quantification of murine cytokine mRNA molecules. AB - A synthetic DNA construct has been developed as a standard molecule whereby murine cytokine mRNA molecules can be quantified by the reverse transcription polymerase chain reaction (RT-PCR). The construct, designated Cytoquant 1, allows the quantification of murine IL-1 alpha, IL-2, IL-3, IL-4, IL-5, IL-6, IL-10, IFN gamma, TNF-alpha, TGF-beta, GM-CSF, CD4, CD8, HPRT and beta-actin mRNA levels. This technique is based on the amplification of a transcribed RNA molecule from Cytoquant 1 as an internal standard control in both the RT and PCR reactions. The quantification data from these analyses are expressed in absolute values, i.e. molecules/cell, which allows the data derived from separate experiments to be compared. In this study, mRNAs encoding beta-actin, IL-10, IFN-gamma and GM-CSF have been quantitated in both Th1 and Th2 cell clones with, and without, stimulation. The quantitative analysis data are highly reproducible and cytokine mRNA concentrations are reflective of restricted cytokine secretion patterns. Furthermore, constitutive cytokine mRNA levels are detectable in resting cells, eliminating the need for exogenous stimulation. The high degree of sensitivity and accuracy make this methodology uniquely suited for the study of T-cell subset cytokine expression in both in vivo and in vitro biological models. PMID- 7477006 TI - Nucleic acid detection using non-radioactive labelling methods. AB - Nucleic acid probe-based assays are now widely used in genetic research, human identification, forensics and in a broad spectrum of clinical assays in the fields of microbiology, haematology/oncology and virology. Labelled probes are used in a variety of assay formats including dot-blots, Southern blots (DNA target), Northern blots (RNA target), Western blots (protein target), in situ hybridization, plaque or colony screening and immobilized arrays on silicon or glass surfaces. Traditionally, the probes used in these assays have a radioactive 32phosphorous label that has a short shelf-life, is dangerous, has high disposal costs and, when labelled to high specific-activity, may be unstable. Extensive efforts to develop alternative labelling techniques have resulted in colorimetric, chemiluminescent and fluorescent assay formats. This review summarizes the properties desired in a probe, describes the advantages and disadvantages of the different non-radioactive labelling strategies, and illustrates examples of probe-based assays in which detection is facilitated by imaging samples using a general purpose fluorescence scanner. PMID- 7477007 TI - Identification of a patient with Streptococcus pneumoniae bacteremia and meningitis by the polymerase chain reaction (PCR). AB - A polymerase chain reaction (PCR) assay based on the penicillin-binding protein gene PBP2B identified the presence of DNA specific for Streptococcus pneumoniae in the serum and CSF of a patient with culture-proven bacteremia and meningitis. Positive signals were seen to dilutions of 1:125 and 1:390,625 for the blood and CSF specimens, respectively. Potential advantages of PCR over conventional culture include exquisite sensitivity, faster results and the ability to identify the organisms by the presence of species-specific DNA even in patients pretreated with antibiotics. PMID- 7477008 TI - Species-specific 18S rRNA gene amplification for the detection of P. falciparum and P. vivax malaria parasites. AB - Based on the sequence diversity of the Plasmodium 18S ribosomal RNA (rRNA), we designed oligonucleotide primers for polymerase chain reaction (PCR) to yield different size fragments for P. falciparum and P. vivax. The primers for the PCR procedure were chosen such that the 5' primer was Plasmodium-conserved while the 3' primers were species-specific. Using primer cocktails and cloned plasmid DNAs containing the 18S rRNA genes or parasite genomic DNA as targets, we show that the PCR procedure yields 1.4-kb and 0.5-kb DNA fragments for P. falciparum and P. vivax, respectively. Limited field testing of this procedure demonstrated the utility of a ribosomal gene based species-specific malaria diagnosis. PMID- 7477009 TI - Development of a species-specific polymerase chain reaction assay for Gardnerella vaginalis. AB - The nucleotide sequence of the region between the 16S and 23S rRNA genes of the facultative anaerobic bacterium Gardnerella vaginalis has been determined, together with the 5' proximal 500 nucleotides of the 23S rRNA gene. Regions suited for the development of specific, probe-confirmable polymerase chain reaction (PCR) assays were selected. PCR assays were evaluated with respect to sensitivity and specificity, the latter in comparison with a number of G. vaginalis reference strains and closely related species like Bifidobacterium spp. In an initial diagnostic study it appeared that the PCR test detected G. vaginalis in 40% of women irrespective of their clinical status. Ten out of 11 patients suffering from bacterial vaginosis as defined on the basis of clinical parameters were carrying G. vaginalis. PMID- 7477010 TI - Fluorescence-based DNA minisequence analysis for detection of known single-base changes in genomic DNA. AB - We describe a rapid, automated method for direct detection of known single-base changes in genomic DNA. Fluorescence-based DNA minisequence analysis is employed in a template-dependent reaction which involves a single nucleotide extension of an oligonucleotide primer by the correct fluorescently-tagged dideoxynucleotide chain terminator. Detection following electrophoresis on denaturing acrylamide gels is facilitated by alkaline phosphatase treatment of reaction products after extension followed by isopropanol precipitation of the dye-tagged, single-base extended primer to remove unincorporated deoxynucleotides. Fluorescence analysis of the incorporated dye tag reveals the identity of the template nucleotide immediately 3' to the primer site. This technique does not require radioactivity or biotinylated PCR product, relies on the incorporation of a single dideoxynucleotide terminator to extend the primer by one nucleotide and takes advantage of the sensitivity of fluorescent terminators developed for automated DNA sequence analysis. As a demonstration, we have applied the assay to human genomic DNA for detection of the sickle mutation in the beta-globin gene, and have also examined feasibility for simultaneous delineation using a multiplex like strategy in a single gel-lane of some of the most common beta-thalassemia mutations in the Mediterranean basin. PMID- 7477011 TI - Allele and genotype frequencies of eight DNA polymorphisms in the Italian population. AB - We report the allele and genotype frequencies in Italians of eight unlinked commonly utilized polymorphic loci: HUMTH01, HUMFES/FPS, HLA-DQA1, LDLR, GYPA, HBGG, D7S8 and GC. Genomic DNA from at least 100 individuals was amplified by PCR and typed after electrophoresis (HUMTH01, HUMFES/FPS), or reverse dot-blot (HLA DQA1 and the other five loci). The allelic frequencies determined were compared with available population data. These results are useful for population and individual identification studies. PMID- 7477012 TI - The frequency of the mitochondrial aldehyde dehydrogenase I2 (atypical) allele in Caucasian, Oriental and African black populations determined by the restriction profile of PCR-amplified DNA. AB - The aldehyde dehydrogenase I (ALDH I) gene codes for a mitochondrial enzyme which plays a major role in hepatic alcohol detoxication. It has been related to alcohol flushing in Orientals bearing the atypical ALDH I2 gene. The variant protein results from a lysine for glutamate substitution at position 487 (G-->A change in exon 12). A procedure for ALDH I2 detection consisting in a differentiation between the 'atypical' allele and the 'wild' allele has been improved through PCR and subsequent MboII digestion. Blood samples collected on anticoagulant or directly absorbed on blotting paper were used for DNA amplification in the presence of two specific oligonucleotidic primers, each one able to incorporate a restriction site in the amplimer. After MboII digestion, PCR products were separated by polyacrylamide gel electrophoresis and then visualized with ethidium bromide. This technique permits a rapid and non radioactive detection of atypical ALDH I2 on a PCR product without the use of allele specific oligonucleotides. It was applied to the study of ALDH I2 allele frequency in random population samples of three ethnic groups: Caucasians, Orientals and African blacks. PMID- 7477014 TI - Molecular screening of genetic defects with RNA-SSCP analysis: the PKU and cystinuria model. AB - RNA single-strand conformation polymorphism (rSSCP) is a recently developed method for detecting genetic defects. This technique requires DNA amplification with a polymerase chain reaction making use of one T7 promoter-containing primer. Amplification products are subsequently transcribed in vitro and the labelled transcripts are analysed for single-strand conformation changes. rSSCP has been applied to mutation screening of the phenylalanine hydroxylase gene and rBAT cDNA, from PKU and cystinuric patients, respectively. Experimental evidence shows that 83% and 86% of screened PKU and cystinuric mutations, respectively, give rise to detectable rSSCP signals. Thus, results obtained show that RNA single strand conformation polymorphism analysis is generally applicable and is a suitable technique for detecting genetic disease causing mutations, both in basic research and in clinical practice. PMID- 7477013 TI - Comparison of heteroduplex and single-strand conformation analyses, followed by ethidium fluorescence visualization, for the detection of mutations in four human genes. AB - Non-isotopic DNA single-strand conformation analysis and heteroduplex analysis by ethidium bromide fluorescence visualization (SSCAE and HAE, respectively) were compared for the detection of 15 different naturally occurring mutations in 15 different DNA samples. The mutations included single nucleotide transitions, transversions and deletions, in CFTR (cystic fibrosis transmembrane conductance regulator), COL4A5 (collagen type IV alpha 5 chain), HEXB (hexosaminidase B), and COL1A2 (collagen type 1 alpha 2 chain) genes, responsible for diseases of medical interest. Genomic DNA from peripheral blood leukocytes or cDNA from reverse transcribed fibroblast mRNA were amplified by polymerase chain reaction (PCR), and then analysed by two SSCAE and one HAE protocol. Fourteen out of 15 mutations (93%) were detected with one or the other method. HAE was more sensitive than SSCAE for the larger products (257-426 bp). The only undetected mutation was then identified with the use of a different primer, located farther from the mutation was then identified with the use of a different primer, located farther from the mutation site, thus increasing the combined efficiency of the two methods to 100%. We believe that combined use of SSCAE and HAE is a good, cheap and safe approach for mutation screening in a human gene. PMID- 7477016 TI - Reverse transcriptase-polymerase chain reaction (RT-PCR): a sensitive method to examine basic fibroblast growth factor-induced expression of the early growth response gene-1 (egr-1) in human umbilical arterial endothelial cells. AB - Immediate-early genes are expressed upon growth and differentiation in a large variety of cells and species. In the present study we investigated the effect of basic fibroblast growth factor (bFGF) on early growth response gene-1 (egr-1) mRNA expression in human umbilical arterial endothelial cells (HUAEC). The detection of this gene in HUAEC was performed by Northern blotting and by reverse transcriptase-polymerase chain reaction (RT-PCR). For RT-PCR specific primers for egr-1 and glyceraldehyde-3-phosphate-dehydrogenase (GAPDH) were constructed and PCR conditions were optimized. bFGF induced a time- and concentration-dependent increase of egr-1 expression. Maximal expression occurred within 30 min of stimulation with bFGF at a concentration of 50-100 ng ml-1. RT-PCR gave highly reproducible and specific results. The comparison of both methods showed comparable results but a higher sensitivity for RT-PCR in detecting the egr-1 mRNA. RT-PCR is an excellent method for detecting the expression of egr-1 mRNA in HUAEC. PMID- 7477015 TI - Fine mapping of randomly distributed multiple deletions of mitochondrial DNA in a case of chronic progressive external ophthalmoplegia. AB - Multiple deletions of mitochondrial DNA have been detected by Southern blotting in the skeletal muscle of a 42-year-old woman with chronic progressive external ophthalmoplegia. A PCR method, using several combinations of primers covering the whole mtDNA as well as sequence analysis, disclosed the wide spectrum of these multiple deletions differing in size, location and sequence at the breakpoint junction. Most involved the major region between the two replication origins. However, three deletions affected the minor region and lacked either the light strand origin of replication or the heavy strand promoter. These data suggest an impairment of mtDNA replication leading to illegitimate recombination and extensive damage of mtDNA. PMID- 7477017 TI - Identification of the major North American bluetongue viruses using nucleic acid amplification techniques. AB - A set of primers (BTV-pr1/2) were selected that hybridized to the VP3 gene of the major North American serotypes of bluetongue virus (BTV). Polymerase chain reaction (PCR) testing yielded positive results from specimens of major North American BTV isolates (serotypes 10, 11, 13 and 17) propagated in Vero cells. In addition, PCR assays were positive from samples of all other BTV serotypes, except BTV-16; however, an alternative primer pair (BTV-prN1/N2) was devised for amplification of this serotype and the major North American BTV serotypes. PCR products were not evident following amplification of related viruses, epizootic haemorrhagic disease virus (EHDV) serotypes 1 or 2, in either PCR test. In addition, slight modification of the nucleic acid extraction method allowed for the amplification of BTV template from ovine and cervine blood, but not from the respective control blood samples. Restriction endonuclease analysis (REA) using AluI and TaqI discriminated the PCR products of BTV serotypes 10, 13 and 11/17. Identification of BTV-11 and -17 was accomplished by PCR product nucleotide sequencing. Thus, using a single gene region (VP3), nucleic acid amplification methods were devised for expeditious serogroup-specific detection of all BTV serotypes and identification of individual North American BTV nucleotypes, which is expected to prove valuable for disease control strategies and retrospective epidemiological analyses. PMID- 7477018 TI - Detection of African horse sickness viruses by dot-blot hybridization using a digoxigenin-labelled probe. AB - In order to develop a non-radioactive dot-blot hybridization assay, for the detection of African-horse sickness virus (AHSV), genome segment 7 from 9 serotypes was amplified by RT-PCR. The resulting PCR products were denatured, immobilized on nylon membranes and then hybridized to a non-radioactive digoxigenin-labelled probe. This probe (265 bp in length) was generated by nested PCR using genome segment 7 of AHSV, serotype 4 as a template. The dot-blot was visualized by chemiluminescence. Positives were obtained from the PCR products amplified from all 9 AHSV serotypes, but not from any other equine virus or orbivirus isolates. The sensitivity and specificity of this probe, together with the simplicity and rapidity of this technique, suggest that a non-radioactive dot blot assay may be useful as a method for the routine and rapid diagnosis of viral infections. PMID- 7477019 TI - Development of specific DNA probes and their usage in the detection of Plasmodium vivax infection in blood. AB - The application of nucleic acid probes, in the detection of pathogenic micro organisms, has become an integral part of diagnostic technologies. In this study, Plasmodium vivax-specific DNA probes have been identified by carrying out genomic subtractive hybridization. In this approach, the recombinant clones from a P. vivax genomic library are screened with radiolabelled human and P. falciparum DNA. The colonies which react with labelled P. falciparum and human DNA are eliminated and those which do not produce any autoradiographic signal have been subjected to further screening procedures. Three P. vivax specific DNA probes have been obtained by these repeated screenings. Further analyses indicate that these probes are specific and sensitive enough to detect P. vivax infection in clinical blood samples when used in a non-radioactive DNA hybridization assay. PMID- 7477020 TI - Specific detection of Campylobacter concisus by PCR amplification of 23S rDNA areas. AB - The phenotypic detection of Campylobacter concisus, a species of considerable genomic and phenotypic heterogeneity, has proven to be rather tedious in the past. Although alternative methods like DNA:DNA hybridization, immunotyping or whole-cell protein electrophoresis are valuable for the specific detection of C. concisus, they are too laborious to be performed in routine settings. Hence a simple Campylobacter concisus-specific PCR assay was developed, based on a target sequence which comprises the most variable areas of 23S rDNA. The PCR assay was successfully evaluated on a broad selection of C. concisus strains and phylogenetically related bacteria. PMID- 7477021 TI - Detection of cytomegalovirus using 'boosted' nested PCR. AB - As a part of a study of an outbreak of CMV infections in a neonatal care intensive care unit, a modified nested PCR was developed for detection of CMV DNA in clinical specimens. Standard nested PCR involves a critical step; passage of PCR products from the first reaction round to the second round. We have adapted a 'boosted' nested PCR which implies amplification in one single step, thus reducing the contamination problems. Nasopharyngeal aspirates and urine samples from patients with perinatal CMV infections, breast milk from some of their mothers, amniotic fluids, urine samples and lymphocytes from seropositive healthy adults were examined by PCR and culture. In the total of 614 of clinical specimens, the PCR test yielded positive results in 51 samples from 14 patients, whereas CMV was isolated in 25 samples from 11 cases only. All samples from healthy individuals were negative. CMV DNA was detected in all culture-positive samples, but all samples from healthy adults were negative. 29/68 culture negative specimens were positive by PCR. No cross-reactivity to other herpes viruses or to human DNA was observed. Our findings show a high sensitivity and a high specificity of the 'boosted' nested PCR. We conclude that the described PCR method can be used for the rapid detection of CMV in clinical specimens with a greatly reduced risk of contamination, and it has proved to be a very useful tool in diagnostic work. PMID- 7477022 TI - Direct detection of Shiga-like toxin-producing Escherichia coli in ground beef using the polymerase chain reaction. AB - We recently reported the development and assessment of a technique for the detection of Shiga-like toxin-producing Escherichia coli (SLTEC) using the polymerase chain reaction (PCR) and a digoxigenin-11-dUTP-labelled DNA probe. This technique has now been adapted for the direct identification of SLTEC in ground beef. Ground beef homogenates were diluted 1000-fold to reduce the concentration of components which inhibit the thermostable polymerase. Assessment of four different ground beef samples using the PCR detection technique revealed that fat content was a major inhibitory component. As few as 30 SLTEC ml-1 of a ground beef homogenate were detected using the PCR technique, although it was necessary to enrich six of the samples for positive detection. These findings indicate that the PCR detection technique is suitable for the identification of SLTEC directly from contaminated ground beef without isolation of the bacterium or purification of its DNA. PMID- 7477023 TI - The sensitive detection of fluorescently labelled PCR products using an automated detection system. AB - The polymerase chain reaction plays a central role in many detection assays and methods to improve the sensitivity and specificity of these detection systems are constantly being explored. In this study we investigated the use of an automated laser fluorescent system (ALF) in the context of DNA-based diagnostics for pathogenic bacteria. PCR products were generated using species-specific primer sets, one of which was labelled with a 5' fluorescein. PCR products with a fluorescent label were detected on line with an ALF DNA sequencer and the sensitivity of detection was found to be comparable to that for DNA probe hybridization with a radioactive probe. The technology was successfully applied to the detection of Mycobacterium tuberculosis supplemented into sputum samples and to the detection of listeria in paraffin-embedded tissue samples. PMID- 7477024 TI - Development of an efficient PCR method for toxin typing of Actinobacillus pleuropneumoniae strains. AB - A method has been developed which allows the determination of the activator, the structural and the secretion genes of the three toxins ApxI, ApxII and ApxIII in Actinobacillus pleuropneumoniae in only two PCR reactions. The oligonucleotide primers were designed to amplify a significant part of the activator and structural genes apxICA, apxIICA and apxIIICA together in a single PCR reaction giving amplification products which differ in length, in order to be clearly separated by agarose gel electrophoresis. Variations in the apxIA and apxIIIA genes which were found in different serotypes were taken into account in the design of the primers to give a uniform amplification product for both variants of the apxIA and the apxIIIA genes. The secretion genes apxIBD and apxIIIBD are also detected in a single PCR reaction containing two pairs of oligonucleotide primers which yield two differently sized fragments to differentiate between apxIBD and apxIIIBD genes. The reference strains of A. pleuropneumoniae serotypes 1-12 and 104 field strains representing all serotypes obtained from various laboratories worldwide were analysed for their content of apx genes. The two PCR reactions give toxin gene patterns which are characteristic for different groups of serotypes in A. pleuropneumoniae and allow the rapid differentiation of five toxin type groups, group 1 including serotypes 1, 5a, 5b, 9 and 11, group 2 including serotypes 2, 4, 6, 8, group 3 with serotype 3, group 4 with serotype 7 and 12 and group 5 with serotype 10. The method enhances and facilitates differentiation of A. pleuropneumoniae strains for diagnostics and epidemiology and allows the detection of serotypes with atypical toxin patterns. PMID- 7477025 TI - Identification of two novel mutations (296 + 1G-C and A46D) in exon 2 of the CFTR gene in Greek cystic fibrosis patients. AB - Two novel CFTR mutations were detected in Greek cystic fibrosis patients. One was a missense mutation, A46D, and the other a splice mutation, 296 + 1G-C. Neither was detected on normal chromosomes. PMID- 7477026 TI - [RSV infection. International symposium from the 89th annual meeting of the German Society for Pediatrics. Wurzburg, September 9, 1993]. PMID- 7477027 TI - Antigenic variability of Shigella flexneri serovars 1-5. AB - Data on the genetic basis of the classification of Shigella flexneri serological variants 1-5 are presented. Subserovars "a" are related to monolysogenic variants of the basic lipopolysaccharide (LPS) structure 0 antigen 3,4, "b" to bilysogenic ones. A scheme of their antigenic variability, with regard to loss of one or both prophages is presented. This scheme helps differentiate between antigenic variability and mixed or superinfections. Recent reports confirming our previously published suggestion to exclude serovar 6 (Shigella newcastle) from Shigella flexneri are analyzed. We propose that antigenic variability of S. flexneri 1-5 results from lysogenization of naturally occurring strains. The possibility of consecutive lysogenization of S. flexneri y (-:3,4) by bacteriophages 6 and 7 has been shown, as exemplified by the circulation of a previously unknown subserovar IV:7,8. PMID- 7477028 TI - [Immune reactions and mechanisms in classical swine fever]. AB - The authors review published data on the immune response in classical swine fever infection. Characteristics of the neutralizing antibodies, cytolytic reactions directed at elimination of infected cells, lymphocyte proliferation, and delayed type hypersensitivity are discussed. Problems of potential theoretical and practical interest are formulated. PMID- 7477029 TI - [Polymerase chain reaction and ligase reactions. Principles, traditional methods, and innovations]. AB - The polymerase chain reaction (PCR) is a favorite among the diagnostic DNA probes. Besides the traditional PCR scheme, the author considers the new strictly quantitative modifications representing a combination of PCR with semiautomated immunoassay, this simplifying the practical use of the method. Errors most frequently occurring in PCR are discussed. The ligase chain reaction, more rapid than PCR, and permitting one to obtain a billion of DNA molecule copies in 30 min, is described, and prospects of its use for rapid diagnosis shown. PMID- 7477030 TI - The effect of cell density on influenza virus replication in CV-1 cells. AB - The number of hemadsorbing cells in CV-1 cell monolayers infected with influenza A virus was higher in semi-confluent cultures than in dense contact-inhibited monolayers. The level of virus-specific protein synthesis as well as the accumulation of virus progeny were also inversely correlated with cell density. At a high multiplicity of infection the majority of these cells in dense monolayers did not express HA on the cell surface, did not synthesize virus specific proteins and survived at least 96 hours after infection. However, these cells developed a partial shut-off of cell protein synthesis and could not be efficiently superinfected. Analysis of the data of virus protein synthesis, progeny virus accumulation, and the assessment of hemadsorption suggest that CV-1 cells in dense monolayers can be infected at a sufficiently high multiplicity of infection, but only a fraction of the infected cells is capable of amplification of virus protein synthesis and progeny virus accumulation. No cell-density effects of comparable extent could be observed in MDCK cells. PMID- 7477031 TI - [Essays on modern molecular genetics. From a lecture course for students of the biology department of Moscow State University]. PMID- 7477032 TI - [Cloning fragments of the RNA polymerase gene of an attenuated variant of the foot-and-mouth disease virus A22]. AB - The cDNA fragments complementary to RNA-polymerase gene and 3'-untranslated genome region of attenuated foot-and-mouth disease virus strain A(22)645 have been synthesized and cloned into a plasmid vector pUC19 in E. coli JM109. The cloned cDNA fragments were characterized as to their size, orientation towards the plasmid, and localization in the virus genome. Restriction maps for complete gene and two cDNA clones were constructed. PMID- 7477033 TI - [Determinant of plasmid pBS221 resistance to potassium tellurite]. AB - The genes regulating resistance to potassium tellurite were cloned from plasmid pS221 with a broad host range. The studied DNA fragment encodes for the synthesis of two polypeptides with molecular mass of 38 and 43 kD. Metal tellurium crystals were localized in the periplasm of Pseudomonas aeruginosa ML4262 (pBS221) cells by electron microscopy. Results of colony hybridization permit us to propose that the cloned genes of potassium tellurite resistance have sites of homology with plasmids RP4(IncP), R27 (IncH), pBS38, pBS79, and R931(IncP-2). PMID- 7477034 TI - [Adhesins from Vibrio cholerae: phenotypic analysis and genetic control of synthesis]. PMID- 7477035 TI - The established Yersinia pestis biovars are characterized by typical patterns of I-CeuI restriction fragment length polymorphism. AB - The genomes of three main biovars of Yersinia pestis were subjected to restriction fragment length polymorphism analysis using I-CeuI endonuclease. I CeuI which is encoded by a mobile intron in Chlamydomonas engamenans recognizes a 25-bp site in the ribosomal RNA rrl gene and cuts DNA of most representatives of Enterobacteriaceae into seven fragments corresponding to the presence of seven rrn-operons. Glycerol-positive Y. pestis strains (biovars antiqua and mediaevalis) contain seven ribosomal operons which can be recognized by I-CeuI endonuclease. However, glycerol-negative strains of Y. pestis biovar orientalis expose only six restriction sites for I-CeuI. The restriction fragment length polymorphism patterns obtained with I-CeuI make it possible to distinguish between three biovars of Y. pestis. Use of another rare cutting restriction enzyme, Bln/I, permits differentiation between pigment-adsorbing and avirulent non-pigment-adsorbing Y. pestis. Still, due to homologous recombination between the two copies of IS 100 insertion sequence bracketing the pgm-locus, the mechanism of deletions in the pgm-locus seems to be confined only to strains of biovars antiqua and mediaevalis, and can be different in Y. pestis strains of biovar orientalis. The I-CeuI restriction patterns of two Yersinia strains isolated within a ten-year period in the port of St. Petersburg and originally identified as Y. pseudotuberculosis 01 turned out to be related to typical representatives of Y. pestis biovar antiqua. These strains could be exported from the same source or circulate among Rattus norvegicus population of the port as non-pigment-adsorbing avirulent immunogenic clone. PMID- 7477036 TI - [Synopsis of modern molecular genetics from a lecture course for students of the biological department of Moscow State University]. PMID- 7477037 TI - [Phenotypic analysis of two morphologically different types of Vibrio cholerae 0139 colonies]. AB - A clinical isolate Vibrio cholerae P16064 serogroup 0139 was shown to produce two different kinds of colonies: opaque encapsulated and translucent devoid of capsule. Low virulence of translucent colonies seems to be due to not only loss of capsule which determines serum resistance, but also to decreased expression of genes controlling the motility, and production of protease and mannose-sensitive adhesion pili. Analysis of the lysogenic properties of the two types of colonies permitted us to propose that simultaneous spontaneous alteration of capsule production and the above-mentioned virulence factors in translucent colonies may be caused by a temperate phage in turbid clones of strain P16064. PMID- 7477039 TI - [Apoptosis in the African swine fever virus-swine mononuclear phagocyte system]. AB - Routine electronmicroscopic examination of interactions between African swine fever virus and mononuclear phagocytes showed signs of apoptosis as a morphological phenomenon and form of cellular death in an infected culture treated with defective interfering virus particles and in target cells under the effect of virus specific cytotoxic T-lymphocytes. The morphology of induced apoptosis in the aforesaid virus-cell system is illustrated and described in detail. PMID- 7477038 TI - [Localizing the major peptides of African swine fever virus and virus-associated enzymes in the virion structure]. AB - African swine fever virus polypeptides with molecular weight of 120, 78, 69, 59, 56, 45, 39, 28, 26, 24, 16, and 14 kD are the major proteins in the purified virions, as shown by electrophoresis and immunoblotting. A mixture of proteases and pancreatic lipase hydrolyzed the polypeptides of 120 and 78 kD in viral preparations at low concentrations of enzymes, polypeptides of 69, 56, 45, 39, 28, and 14 kD disappeared after treatment with this mixture at medium concentrations, and 26 kD polypeptide was eliminated at a high concentration of the enzymes. The 21 kD polypeptide which did not react with the specific antiviral serum in immunoblotting was not hydrolyzed by proteases contaminating lipase. Treatment with triton X-100 and ether boosted the activity of DNA dependent RNA-polymerase, whereas treatment with ether followed by resedimentation markedly decreased polymerase activity in the resultant sediment. Treatment with diethyl ether did not influence the activity of virus-associated ATPase, which was partially resistant to denaturating organic solvents acetone and chloroform-methanol mixture. Our findings and published data permitted us to propose a schematic arrangement of viral polypeptides and enzymes in the virion structure. PMID- 7477040 TI - [Polymeric forms of the influenza virus nucleoprotein]. AB - Two high-molecular-weight types of in vivo formed nucleoproteins (NP) capable of reacting with anti-NP monoclonal antibodies were detected in continuous MDCK cells and primary CEFs, along with the monomeric virus NP. High-molecular NP were stable at 37 degrees C, despite the presence of dodecyl sulfate and B mercaptoethanol, but were converted to monomeric NP at 37 degrees C in the presence of 6 M urea, 1 M NaCl, and at acid pH. Hence, high-molecular-weight NP may be the polymeric forms of NP, which are probably stabilized by noncovalent bonds. The kinetics of NP-polymers formation suggests that NP-monomers are their precursors and NP-polymers are formed rapidly after monomeric NP molecules synthesis. Both forms of NP-polymers are detected in the sedimented and soluble fractions of not only cell lysate, but in the extracellular maintenance medium as well. NP-polymers are resistant to RNAse and are more resistant to protease than NP-monomers. PMID- 7477041 TI - An in vitro binucleated blocked hepatic cell technique for genotoxicity testing in fish. AB - The main objective of this work was to develop and use a binucleated method which depends on the cytokinesis blocking of hepatic cells in fish to enable the scoring of micronuclei in cells that have completed only one division after genotoxic insult. The use of this novel technique in a study of the influence of some pollutants in vitro (selenium, mercury, methylmercury, and their mixtures) allows evaluation of the risk from exposure to these water pollutants under field and laboratory conditions. The frequencies of micronuclei in trout hepatic cells in vitro were elevated in a dose-dependent manner when compared to the relevant controls. Addition of Se(IV) reduced the frequencies of micronuclei in treatments with both forms of mercury. Inorganic mercury was found to be partly methylated when exposed to trout liver cells in vitro. The potential contribution of the fish binucleated hepatic technique to the toxicity assessment of the chemicals investigated is discussed. PMID- 7477042 TI - Induction of deletion mutations by methoxyacetaldehyde in Chinese hamster ovary (CHO)-AS52 cells. AB - We have reported previously that methoxyacetaldehyde (MALD), a metabolite of 2 methoxyethanol, induces gpt gene mutations in Chinese hamster ovary (CHO)-AS52 cells but not hprt gene mutations in the standard CHO-K1-BH4 cells. In addition, MALD induces chromosome aberrations in both CHO cell lines. The data presented suggest that MALD induces deletion-type mutations. In this study, we analyzed MALD-induced CHO-AS52 mutants for deletion-type mutations using the nested polymerase chain reaction (nested-PCR) assay. Spontaneous CHO-AS52 mutants are used as untreated control. Ethylnitrosourea (ENU)-induced CHO-AS52 mutants are used as negative control for multilocus deletions since ENU is a potent inducer of point mutations. The results show that the frequency of MALD-induced mutants containing total deletion of the gpt gene is 42.4% which is 2.3-fold higher than that from spontaneous mutants (18.6%). The frequency of ENU-induced deletion mutation is 3%. The data substantiate our hypothesis that MALD induces major deletion mutations. PMID- 7477044 TI - Effects of aluminum chloride on the nucleus and nucleolus in root tip cells of Hordeum vulgare. AB - The cytotoxic effects of aluminum chloride at different concentrations (1 x 10( 4)-5 x 10(-2) M) on the nucleus and the nucleolus in root tip cells of Hordeum vulgare were investigated using the carbol fuchsin staining method and the silver staining technique separately. Results showed that aluminum chloride could induce nuclear aberrations comprising elongated, irregular, ruptured and fractured nuclei and micronucleus formation. After the treatment with aluminum, it was observed that the nucleolar material was extruded from the nucleus into the cytoplasm. The nucleolar material in the cytoplasm was disintegrated, forming silver-stained particulate material, which occurred in both the inner root meristem cells and the root cap cells. The possible mechanism of the aluminum toxicity on the nucleus and nucleolus is briefly discussed. PMID- 7477045 TI - Chromosomal radiosensitivity of lymphocytes from skin cancer-prone patients. AB - Frequencies of spontaneous and radiation-induced chromosome aberrations were documented in lymphocytes from patients with basal cell nevus syndrome (BCNS) and from those with epidermodysplasia verruciformis (EV). Cells were irradiated with single or double doses of gamma-rays or UV light. For the double dose irradiation protocol, the two doses were separated from each other by 60 min. After irradiation of cells with 4 or 2 + 2 J/m2 UV light, lymphocytes from seven BCNS patients had a similar number of aberrant cells compared with normal controls, while cells from three EV patients showed a significantly increased number of cells containing aberrations, particularly chromatid-type aberrations, compared to controls (p < 0.03, Student's t-test). Similar results were observed whether the cells were irradiated with single or double doses of UV light. In addition, lymphocytes from EV patients had significantly lower mitotic indices than controls whether the cultures were irradiated with UV light or unirradiated (p < 0.05; Student's t-test). After irradiation with 100, 100 + 100 or 50 + 50 cGy gamma-rays, the mean chromosome aberration frequencies from nine BCNS patients and two EV patients were not significantly different from the controls. The spontaneous chromosome aberration frequencies were similar for the patients and the controls. Our data indicate that BCNS and EV lymphocytes irradiated with gamma-rays have a normal DNA repair response. However, cells from EV patients have an abnormal repair response to UV light induced DNA damage. This abnormality is probably caused by deficiency in DNA repair and it is consistent with the patients' sensitivity to UV light induced cutaneous squamous cell carcinomas. PMID- 7477043 TI - Supernatant medium from UV-irradiated cells influences the cytotoxicity and mutagenicity of V79 cells. AB - It is known that UV light induces the secretion of some proteins into the extracellular medium. We have carried out experiments to study how the supernatant medium from UV-irradiated cells affects the cytotoxicity and mutagenicity of V79 cells exposed to different damaging agents. So we exposed exponentially growing cells to 20 J/m2 of UV light and then harvested the supernatant medium after 22 h. This supernatant medium was then used to treat a fresh batch of cells for 2 h. After the treatment with this supernatant medium the cells were subsequently exposed to UV light, gamma-rays, hydrogen peroxide or MNNG. We found that exposure to this medium had a protective effect on the survival levels for UV light, gamma-rays and hydrogen peroxide while MNNG-induced killing remained unaffected. With UV light and gamma-rays we found that mutation induction at all doses was increased. Cycloheximide could inhibit this protection and the increase in mutation frequencies was also suppressed. The results indicated a protective role for the UV-induced factor(s). They were probably involved directly or they triggered repair process(es) that were related to oxidative stress. PMID- 7477046 TI - Methodological aspects of the white-ivory assay of Drosophila melanogaster. AB - The white-ivory somatic assay of Drosophila melanogaster was developed to detect genotoxic agents which induce loss of a tandem duplication. Although the mechanism of this loss is not known, some suggestions point to intrachromosomal recombination as the main reversion mechanism. Since the few papers published to date on this assay present controversial methodologies, prior to a larger study of chemicals with different mechanisms of action, we have carried out an analysis to optimize some conditions of this assay. For this purpose, we have used three different strains and four well characterized mutagenic chemicals: N-ethyl-N nitrosourea (ENU), methyl methanesulfonate (MMS), ethyl methanesulfonate (EMS) and hexamethyl phosphoramide (HMPA). The results obtained allow us to conclude that: (i) the best strain for this assay is C(1)DX,y,f/Dp(1:1:1:1)wi,y2, although the use of strain FM6,l(1)66a/Dp(1:1:1:1)wi,y2;st/st could be considered for some mechanistical studies; (ii) developmental reasons make it necessary to use as estimate of reversion frequency the proportion of eyes showing at least one spot; (iii) reversion frequency cannot be used as estimate of mutation efficiency, neither can spot size evaluate time of spot induction; (iv) the four chemicals clearly induce loss of the wi duplication; according to their activities they rank ENU > HMPA > MMS approximately EMS. PMID- 7477047 TI - Evaluation of micronuclei in exfoliated urothelial cells and urinary thioether excretion of smokers. AB - Mutagens are present in large quantities in the urine of cigarette smokers, thus, their urothelial cells may represent a possible target for absorbed and excreted mutagens. Our aim is to validate the micronucleus (MN) test in exfoliated urothelial cells obtained from urine samples of cigarette smokers. The urinary thioether (UT) test is also carried out on the same individuals in order to find out whether there is any correlation between these two end-points. The mean (+/- SE) MN frequency and UT determination is 1.93 (+/- 0.11)% and 9.71 (+/- 1.61) mmol SH/mol creatinine for 23 smokers, and 0.66 (+/- 0.05)% and 4.20 (+/- 0.56) mmol SH/mol creatinine for 20 nonsmokers. Our results show a higher frequency of micronucleated cells (p < 0.001) and higher excretion of UTs (p < 0.05) in smokers as compared to nonsmokers. Concentrations of UTs and MN frequencies increased with tobacco consumption. The MN frequencies showed only a marginal increase, not significant (p > 0.05), after passive smoking compared to nonsmoking values. There was no significant correlation between MN frequencies and UTs, either in smokers (r = 0.164, p > 0.05) or in nonsmokers (r = -0.018, p > 0.05). Our data demonstrate tobacco-induced chromosome damage in bladder tissue consistent with increased risk of cancer at this site among smokers. PMID- 7477048 TI - Do persons living near a uranium processing site have evidence of increased somatic cell gene mutations? A first study. AB - The objective of this study was to examine if individuals living near a uranium processing site have greater mutagenic damage, as measured by three mutagenicity assays, compared with subjects unexposed to any nuclear facilities. The design was a cross-sectional exploratory analysis of 112 subjects; 56 volunteer residents were from within a 5-mile radius of the Fernald Uranium Processing site and 56 'control' subjects were from a geographically separate area unexposed to any known uranium emissions. The groups were constrained to be similar in age and sex composition. The main outcome measures were three human somatic gene mutation assays consisting of the HPRT T-lymphocyte cloning assay to measure 6-thioguanine resistant lymphocytes; the glycophorin A assay to detect the loss of expression of the M or N allele; and the micronucleus assay as a marker of chromosomal damage. The results showed no statistically significant or quantitatively important differences between groups for all three mutagenicity assays; only the unselected cloning efficiency was statistically significantly different between groups (0.42 +/- 0.16 for the Fernald versus 0.35 +/- 0.12 for the comparison groups). In both groups, age was significantly related to HPRT mutant frequency, with a 1.25% rate of increase in mutant frequencies for each 1-year gain of age in the Fernald group and a 1.12% rate of increase in mutant frequencies for each 1-year gain of age in the comparison group. For the micronucleus data, females had a greater mean micronucleus frequency than males. In addition, smokers had an increased mean ln (natural logarithm) HPRT mutant frequency (3.06 +/- 0.14 for current smokers compared with a mean of 2.72 +/- 0.05 for non-current (i.e. never plus former) smokers). Our results are consistent with the previously reported association between sex type and micronucleus frequency, the known relationship between age and T-lymphocyte cloning efficiency and age and HPRT mutant frequency, and verify the wide inter-subject variability for the latter. Finally, we conclude that at a population level, the relationships between current cigarette use and HPRT mutant frequency, and sex type and micronucleus frequency, are stronger than is the association between geographic proximity to a uranium processing site and mutagenic abnormalities. PMID- 7477049 TI - Comparison of lacI and lacZ transgenic mouse mutation assays: an EU-sponsored interlaboratory study. AB - As part of an EU Environment Programme sponsored study, the performance of two transgenic mouse mutation assays has been evaluated in three laboratories using common liver samples. The systems studied were the lacI-(Big Blue) mutation assay, and the GalE- positive selection modified lacZ- (Muta Mouse) assay. The liver samples compared were derived from mice treated with either saline or dimethylnitrosamine (DMN; 14 day recovery). Each assay gave an increased mutation frequency (MF) for the DMN-treated livers when compared to the saline control MFs. Sources of variability in the assays are discussed, and it is concluded that whole liver should be homogenised before DNA extraction, and that concurrent controls should be processed in parallel with test samples. PMID- 7477051 TI - An in vivo/in vitro method for assessing micronucleus and chromosome aberration induction in rat bone marrow and spleen. 2. Studies with chlorambucil and mitomycin C. AB - An in vivo/in vitro system using rat bone marrow cells and spleen cells to assess micronucleus (MN) and structural chromosome aberrations (SCA) simultaneously (Moore et al., 1995) was further developed. In two separate experiments, two rats/dose/experiment were treated i.p. with 0, 5, 10 and 15 mg chlorambucil (CA)/kg or with mitomycin C (MMC) at 0, 1, 2, 4 mg/kg (experiment 1) or 0, 4, 6, and 8 mg/kg (experiment 2) and killed 6 h later. Cultures were then established in the presence of growth stimulants (interleukin-3 and granulocyte-macrophage colony stimulating factor for bone marrow; lipopolysaccharide and concanavalin A for spleen) and cytochalasin B, a cytokinesis inhibitor. Bone marrow cells were harvested 24 h after establishment of cultures, while spleen cells were harvested at 48 h. In addition, spleen cells were concurrently assayed for chromosome aberrations. With the MN endpoint, spleen cells appeared more sensitive than bone marrow cells to the effects of CA due both to a lower background and an increased response. For MMC, bone marrow cells exhibited both a higher background of MN and a greater numerical response than did spleen cells. However, on the basis of a fold-increase over control values, spleen cells were more sensitive than bone marrow cells. In general, the MN endpoint appeared more sensitive than the SCA in spleen cells after treatment with CA or MMC. Thus, the approach described here shows greater potential in detecting genotoxicity. PMID- 7477052 TI - Distributions of nitropyrenes and mutagenicity in airborne particulates collected with an Andersen sampler. AB - Distributions of 1,3-dinitropyrene (1,3-DNP), 1,6-DNP, 1,8-DNP, 1-nitropyrene (1 NP) and mutagenicity in airborne particulates collected in downtown Kanazawa, Japan with an Andersen high-volume air sampler were examined. Mutagenicities of benzene-ethanol extract from particulates were determined by the Ames test using S. typhimurium strains without S9 mix, while concentrations of DNPs and 1-NP were determined by high-performance liquid chromatography (HPLC) using chemiluminescence detection. In the finest particulate fraction (smaller than 1.1 microns), 68% and 75% of the total mutagenicities were observed in TA98 and YG1024 strains, respectively. In the same fraction, 65-82% of three DNPs as well as 84% of 1-NP were observed. Mutagenic contributions of 1,3-DNP, 1,6-DNP, 1,8 DNP and 1-NP in the extract were respectively 0.6, 1.2, 1.8 and 1.6% in the TA98 strain, and 2.5, 5, 9 and 2.1% in the YG1024 strain. PMID- 7477050 TI - An in vivo/in vitro method for assessing micronucleus and chromosome aberration induction in rat bone marrow and spleen. 1. Studies with cyclophosphamide. AB - The mouse micronucleus assay has long been used as an indicator of in vivo genotoxicity. Recently, it was shown that no single protocol is adequate to detect all clastogens. As a first step in developing a potentially more sensitive assay, micronucleus induction by cyclophosphamide (CP) was assessed in an in vivo/in vitro system using rat bone marrow and spleen cells. In each of two independent experiments, two rats/dose were treated i.p. with 0, 20, or 40 mg CP/kg and killed 6 h later. Cultures were then established in the presence of growth stimulants (interleukin-3 and granulocyte-macrophage colony stimulating factor for bone marrow; lipopolysaccharide and concanavalin A for spleen) and cytochalasin B, a cytokinesis inhibitor. Bone marrow cells were harvested and slides prepared 24 h after initiation, while spleen cells were harvested at 48 h. One thousand cells/tissue/group were scored for cell cycle kinetics and 1000 binucleate (BN) cells were scored for micronuclei. In addition, spleen cells were concurrently assayed for chromosome aberrations. A dose-related cell cycle delay was observed in both tissues in both experiments. Bone marrow showed a 6% average background frequency of micronucleated BN cells, while the low dose induced an average of 20%, and the high dose 31%. For spleen, the average control frequency of micronucleated BN cells was 3%, the low dose induced a 40% average frequency, and the high dose 65%. Also in splenocytes, a dose-dependent increase in chromosome aberrations was observed, with an almost 40-fold increase observed over the control value at the high dose. Thus, the in vivo/in vitro approach described here shows great potential in detecting drug induced genotoxicity. Also, spleen appears more sensitive than bone marrow to CP. PMID- 7477053 TI - The transmission rate of the lacI transgene from the Big Blue mouse. AB - Since transgenic mice are being used to analyze somatic and germinal mutation rates in vivo, it is of interest to know to what extent these mice are normal or abnormal in any way. During experiments designed to compare the mutational response of the transgene and an endogenous gene, Big Blue mice hemizygous for the transgene were bred to create a hybrid mouse in which the comparisons could be made. The fraction of these mice that inherited the transgene was 37% rather than the Mendelian expectation of 50%. PMID- 7477054 TI - Inhibitory effect of curcumin on SOS functions induced by UV irradiation. AB - The antigenotoxic effects of curcumin, including the inhibition of SOS induction and mutagenesis by UV light, were investigated in Salmonella typhimurium TA1535/pSK1002 and Escherichia coli K-12 strains. Induction of the SOS gene (umuC) expression was assayed by measuring accumulated beta-galactosidase activity. We found that curcumin blocked umuC induction promoted by UV irradiation in a dose-dependent manner. Also, with another SOS response, Weigle reactivation, we observed that curcumin effectively inhibited phage reactivation by UV irradiation. Furthermore, we tested the effect of curcumin on UV mutagenesis. We showed that mutagenesis induced by UV irradiation was suppressed by the addition of curcumin. Together these results indicate that curcumin acts as an inhibitor of SOS functions including UV mutagenesis. PMID- 7477055 TI - Comparison of sister chromatid exchange frequency in peripheral lymphocytes in lung cancer cases and controls. AB - Cytogenetic alterations have been associated with the occurrence of many cancers. However, limited data exist to address whether increased chromosomal changes in surrogate normal tissue are similarly associated with malignancy. As part of an ongoing case-control study of lung cancer, we have studied the factors that affect sister chromatid exchange (SCE) frequency in lymphocytes from lung cancer patients. Further, we sought to investigate whether the factors that affect SCE frequencies were comparable in lung cancer cases and controls. Cases had newly diagnosed, operable primary lung cancer. Controls were friends and spouses of cases. Detailed information on smoking, family history of cancer, medical history, and environmental and occupational exposures was obtained in an interviewer-administered questionnaire. Intake of antioxidants was also determined through the administration of a validated semiquantitative food frequency questionnaire. Metabolic traits studied included the polymorphic glutathione-S-transferase class mu (GST-mu) and variants of P450 isoenzymes CYP1A1 and CYP 2D6. Overall, 78 cases and 78 controls were included in the analysis. Although there was a small number of lung cancer patients who had never smoked in the study (9% of cases), these patients had higher SCE frequencies than current or former smokers. This suggests that factors associated with genomic instability may also play a role in the pathogenesis of lung cancer. The best fit model for SCE frequency, which had been previously generated from control data alone, included age, gender, smoking, GST-mu, and vitamin A intake. However, when this model was applied to lung cancer patients, smoking was not associated with an elevated SCE frequency. Thus, it is not clear that SCE frequency data in prevalent lung cancer cases and controls are comparable. PMID- 7477056 TI - Frequency of HPRT mutant lymphocytes in a human control population as determined by the T-cell cloning procedure. AB - The T-cell-cloning assay was established to determine the frequency of hypoxanthine-guanine phosphoribosyltransferase (HPRT) mutant lymphocytes in the presence of the selective agent 6-thioguanine in peripheral blood from a human control population. We investigated 44 healthy adults (blood donors) and found a mean mutant frequency of 7.2 x 10(-6) (geometric mean 5.6 x 10(-6). An elevated mean mutant frequency occurred in smokers as compared to non-smokers. However, a statistically significant increase was only observed between female smokers and female non-smokers while there was only a slight difference in the male group. A significant difference in mutant frequency could be found between individuals younger than 35 years and those above 35. But the difference of the mutant frequency with age showed up only among smokers. No significant effect of the gender was observed. Mutant frequency was inversely related to the cloning efficiency. PMID- 7477058 TI - Electrophysiological and urodynamic studies to monitor surgical outcome in children with tethered spinal cords. AB - Children with myelodysplasia often develop tethering of their spinal cord within 2-3 years after correction of their initial birth defect, leading to either neurological or urological dysfunction. Surveillance of these children includes electrophysiologic/urodynamic (EMG/UDS) studies to monitor improvement or deterioration of their neurological function. We reviewed 39 patients who underwent surgical untethering from 1979 to 1994 to determine whether postoperative EMG/UDS studies were useful in assessing outcome after surgery. All patients had a complete neurologic examination and an EMG/UDS prior to and 3 months after surgery. Twenty patients (51%) showed improvement in their EMG/UDS study after untethering, 15 (39%) demonstrated no significant change, and 4 (10%) showed deterioration. Improvement consisted of either return of previously absent motor units, return of sacral reflexes, or improved bladder dynamics. In a further 3 patients, subsequent deterioration on serial postoperative EMG/UDS testing was the first indication of recurrent tethering. We conclude that serial EMG/UDS evaluation in patients at risk for tethering of the spinal cord may serve as a useful adjunct to close clinical observation. PMID- 7477057 TI - Effects of indomethacin and arachidonic acid on sister chromatid exchange induction by styrene and styrene-7,8-oxide. AB - Styrene is converted into styrene-7,8-oxide in human lymphocyte cultures, in a reaction probably mediated by oxyhemoglobin. As a consequence, styrene induces sister-chromatid exchanges (SCEs) in whole-blood lymphocyte cultures without exogenous metabolic activation systems. Another metabolic pathway that could be involved in the metabolism of styrene is cooxidation by prostaglandin endoperoxide synthase (PES). To study the role of PES in the metabolism of styrene, human whole-blood lymphocyte cultures were treated for the entire culture time of 72-h with styrene (0.5 and 1 mM) or styrene-7,8-oxide (50 and 100 microM), in the presence and absence of 75 or 150 microM indomethacin (an inhibitor of PES) and arachidonic acid (substrate of PES). Indomethacin potentiated SCE induction by both styrene and styrene-7,8-oxide; a slight but statistically significant enhancement (16-32%; p < 0.05-p < 0.001) was observed in all treatments with styrene and at 150 microM indomethacin in the case of styrene-7,8-oxide. At 150 microM, arachidonic acid induced a 15-20% suppression (p < 0.01) in SCE induction by both styrene (1 mM only) and styrene-7,8-oxide (100 microM only). Indomethacin or arachidonic acid did not alone influence the frequency of SCEs. The results suggest that PES acts as an inactivation route for styrene and styrene-7,8-oxide in human whole-blood lymphocyte cultures, possibly through PES-mediated binding to glutathione. PMID- 7477059 TI - X-linked recessive bulbospinal neuronopathy: clinical phenotypes and CAG repeat size in androgen receptor gene. AB - Clinical phenotypes and the CAG repeat size of the androgen receptor gene were assessed in 95 Japanese patients with X-linked recessive bulbospinal neuronopathy. There was an age- and duration-dependent deterioration of muscle strength, ADL scores, and plasma creatine kinase levels. However, there was no correlation between the presence of absence of gynecomastia or diabetes mellitus and the age at onset or duration of illness. Correlations were present between the CAG repeat size and the age at onset (P < 0.0001) as well as the presence or absence of gynecomastea (P < 0.05). Muscular weakness and ADL scores were also correlated with the CAG repeat size only when they were adjusted by the age at examination not by the duration of illness. These findings suggest that CAG repeat size is one of the determinant factors of disease progression. However, extensive variation in phenotypic severity in patients with the same size of CAG repeat was present even among the siblings, suggesting that other factors than CAG repeat size influence the phenotypic manifestation. The average gain of CAG repeat size expansion was a 1.4 repeat in paternal transmission which was more unstable than that in maternal transmission, but the magnitude of the expansion in paternal transmission was much smaller than is presently known for other diseases in which CAG repeat expansion is the responsible gene defect. PMID- 7477060 TI - Influence of recording site within the muscle on motor unit potentials. AB - The influence of the recording site on the motor unit potentials (MUPs) was investigated in the brachial biceps muscle of 8 healthy subjects. The MUPs were recorded with a concentric needle electrode and analyzed with a new decomposition EMG program we call multi-MUP analysis. MUPs had shorter durations and smaller amplitudes at superficial recording sites than at deeper sites in the muscle. This is mainly due to the cannula of the concentric electrode, which records a higher potential at superficial recording sites and partially cancels the recorded potentials from the tip in a differential recording. The MUPs had longer durations and higher amplitudes distally than in the middle of the muscle. The longer durations and spike durations are probably due to increased temporal dispersion at a greater distance from the endplate zone. We do not have an adequate explanation for the larger amplitudes distally in the biceps, they may be due to anatomical factors. To increase the diagnostic sensitivity of quantitative MUP analysis the recordings should be obtained from standardized recording sites. PMID- 7477061 TI - Quantitation of heteroplasmy of mitochondrial tRNA(Leu(UUR)) gene using PCR-SSCP. AB - We have devised a novel method for quantitative analysis of the MELAS (mitochondrial myopathy, encephalopathy, lactic acidosis, and stroke-like episodes) tRNA(Leu(UUR)) mutation of mitochondrial DNA using a PCR-SSCP (polymerase chain reaction-single-strand conformation polymorphism) method, and compared the results obtained using the PCR-SSCP method with those obtained using other methods including Southern blotting, last one cycle hot PCR, and conventional PCR-RFLP (restriction fragment length polymorphism). The standard curve obtained using the PCR-SSCP method is linear, with a correlation coefficient of 0.999; it was determined that this method is more accurate than other methods for quantitative analysis. The PCR-SSCP method does not require restriction digestions, thereby avoiding potential problems of partial digestions or heteroduplex formation during PCR. The method is quite simple and should have a broad range of application for quantitation of mutant mtDNAs in various mitochondrial encephalomyopathies. We applied the method for quantitation of mutant mitochondrial DNA carrying a single base substitution in the tRNA(Leu(UUR)) gene in two autopsied cases of MELAS. In both cases, the mutant mtDNA is abundantly present (82-95%) withd little variation among tissues. PMID- 7477062 TI - Vrms/T quantitation: usefulness in patients with neuropathies. AB - This study evaluates the sensitivity of root mean square voltage (Vrms)/turns (T) ratios in neuropathies. Data were recorded with concentric needle electrodes from the biceps brachii, first dorsal interosseous, tibialis anterior, and calf muscles. Recordings were analyzed from 35 normal subjects as well as 24 consecutive patients referred for evaluation of neuropathic disorders. Direct tension measurements were not required. Reasoning from studies in normal subjects in which Vrms/T values were related to measurements of relative tension, the data were recorded during strong muscle contractions of less than 80% of maximum tension as monitored by mean amplitude values. For all 5 muscles, mean Vrms/T values > or = 1.0 calculated from four separate needle sites were significantly more frequent in the patients than controls (p < 0.012-0.0001). There were 13 muscles in which relevant conduction studies were abnormal and Vrms/T > or = 1.0 were the only abnormality of the needle EMG examination. In addition, a Vrms/T of > or = 1.0 was present in 10/14 muscles in which abnormalities might have been expected but where other needle EMG parameters were unremarkable and relevant nerve conductions normal. This study indicates that Vrms/T measurements can add meaningfully to the electrodiagnostic evaluation of neurogenic dysfunction. PMID- 7477064 TI - Thenar motor unit number estimates using the multiple point stimulation technique: reproducibility studies in ALS patients and normal subjects. AB - Thenar motor unit number estimate (MUNE) reproducibility was assessed in 20 patients with amyotrophic lateral sclerosis (ALS) and 16 normal subjects using the multiple point stimulation (MPS) technique. The MUNE was calculated by dividing the thenar compound muscle action potential negative-peak (n-p) area by the mean n-p area of 10 lowest threshold, all-or-nothing, surface-recorded motor unit action potentials. Two trials (test-retest) were performed by the same examiner either on separate days or on the same day with new electrode placements. The mean test MUNE was 43.4 (SD: 35.9, range: 6-145) for ALS patients and 219.4 (SD: 80.8, range: 122-368) for normal subjects. Test-retest MUNE differences were not significant for ALS patients or normal subjects. The test retest correlation coefficient (r) was 0.99 for ALS patients and 0.85 for normal subjects. The mean difference between test-retest values was 10% for ALS patients and 17% for normal subjects. Test-retest reproducibility of the thenar MUNE using the MPS technique is high in both ALS patients and normal subjects. The reliability of the MPS technique in estimating motor unit numbers may make it a useful outcome measure in following the course of patients with progressive lower motor neuron disease, especially those enrolled in experimental drug trials. PMID- 7477065 TI - Myogenic cells derived from rat bone marrow mesenchymal stem cells exposed to 5 azacytidine. AB - The compound 5-azacytidine has been previously shown to convert cells of the rat embryonic fibroblastic cell line, C3H/10T1/2, into myoblasts, adipocytes, and chondrocytes. Rare, resident cells of bone marrow and periosteum, referred to as mesenchymal stem cells, have been shown to differentiate into a number of mesenchymal phenotypes including bone, cartilage, and adipocytes. Rat bone marrow derived mesenchymal stem cells were exposed to 5-azacytidine beginning 24 h after seeding twice-passaged cells into culture dishes. After an exposure of 24 h, long, multinucleated myotubes were observed in some of the dishes 7-11 days later. Cells containing Sudan black-positive droplets in their cytoplasm were also observed. Thus, culture-propagated rat bone marrow mesenchymal stem cells appear to have the capacity to be induced to differentiate in vitro into myogenic and adipocytic phenotypes, although nonmesenchymal cells (rat brain fibroblasts) cannot be so induced. Taken together, these observations provide support for the suggestion that mesenchymal stem cells in the bone marrow of postnatal organisms may provide a source for myoprogenitor cells which could function in clinically relevant myogenic regeneration. PMID- 7477063 TI - Evidence of an abnormal intramuscular component of fatigue in multiple sclerosis. AB - The goals of this study were to investigate muscle fatigue in patients with multiple sclerosis (MS), and to determine the relationships between muscle fatigue, clinical status, and perceived fatigue. The fatigability of the anterior tibial muscle was quantitated in patients and controls during 9 min of intermittent stimulation (used to eliminate central sources of muscle fatigue). During exercise, the decline in tetanic force, phosphocreatine, and intracellular pH was greater in patients than in controls. The compound muscle action potential amplitude did not decrease during exercise, indicating that there was no failure of neuromuscular transmission during fatigue. Thus, the excessive fatigue in MS developed from sources beyond the muscle membrane. Following exercise, the recovery of tetanic force was delayed in patients (a pattern that suggests abnormal excitation-contraction coupling), whereas the recovery of metabolites was complete in both groups. Muscular fatigue was correlated with clinical disability but not with perceived fatigue. These results suggests that fatigue in MS has both central (perception, upper motor neuron dysfunction) and peripheral (impaired metabolism and excitation-contraction coupling) components. PMID- 7477066 TI - Facilitation of motor evoked potentials: timing of Jendrassik maneuver effects. AB - Remote voluntary contraction, such as the classical Jendrassik maneuver (JM), is a procedure routinely used to increase the amplitude of tendon reflexes in the lower limb. In 8 healthy subjects we studied the effects of JM on the motor evoked potentials (MEP) recorded from tibialis anterior muscle, produced by transcranial magnetic stimulation (stimulus output of 5-10% over motor threshold). In this study, JM consisted here of a bilateral violent handgrip, preceding magnetic stimulation from 100 to 50 ms (steps of 100 ms). Compared to the control test, latencies remained unchanged. MEP amplitudes were greatly enhanced with a JM test interval from 200 to 400 ms (170% of control amplitude at 300 ms). We also studied 6 patients with severe alterations of MEPs from tibialis anterior muscle. In each case, JM preceding magnetic stimulation (stimulus output 100%) from 300 ms induced reappearance of response or marked enhancement of amplitude, allowing calculation of central conduction time. Such a technique, which is easy to perform, may be useful in clinical practice to calculate central motor conduction time, where it would otherwise be difficult or impossible. PMID- 7477067 TI - Insulinlike growth factor gene expression in rat muscle during reinnervation. AB - Because insulinlike growth factors (IGFs) support motor axon regeneration, we tested whether the IGF genes expressed during the development of neuromuscular synapses are reexpressed in adult rat muscles during synapse regeneration. Following sciatic nerve crush, IGF-II mRNAs per poly(A)+ RNA, as well as per poly(A)+ RNA per milligram muscle, were significantly up-regulated in denervated relative to intact contralateral gastrocnemius muscles. IGF-II mRNAs were down regulated after the reestablishment of functional neuromuscular synapses, but remained up-regulated when nerves were transected to prevent the reestablishment of synapses. These data are consistent with a model in which the IGF-II gene is reexpressed during regeneration due to loss of nerve-dependent feedback inhibition. There was a slight but significant increase in IGF-I mRNAs per poly(A)+ RNA per milligram muscle, probably as a consequence of muscle atrophy. These results show that IGF-II gene expression is up-regulated in muscle during the reestablishment of synapses. PMID- 7477068 TI - Effects of temperature on motor unit action potentials during isometric contraction. AB - The effect of temperature on motor unit action potential (MUAP) configuration and recruitment was studied using automatic decomposition electromyography (ADEMG) recordings from a concentric needle electrode placed in the first dorsal interosseous (FDI) muscle of 10 normal adult subjects during isometric contraction. Focally cooling the FDI resulted in prolonged MUAP duration (P < 0.001, ANOVA), a finding congruent with those of Buchthal. Focal ulnar cooling at the elbow resulted in the increased MUAP frequency. In contrast to previous studies, there were no significant differences in amplitude or turns. Greater understanding of normal motor unit electrophysiology is necessary to improve diagnostic accuracy of EMG testing. PMID- 7477069 TI - Beneficial versus adverse effects of long-term use of clenbuterol in mdx mice. AB - Long-term administration of the beta 2-adrenergic agonist clenbuterol in mdx mice was used to test the hypothesis that increasing contractile protein content in skeletal muscle will decrease the progression of muscular dystrophy. C57BL/10SNJ (control) and dystrophic (mdx) mice were given clenbuterol (1.0-1.5 mg/kg body weight/day) in the drinking water. Ventilatory function and morphological and functional characteristics of soleus (SOL) and diaphragm (DIA) muscles were evaluated. Clenbuterol administration was associated with increased SOL muscle weight, and SOL muscle weight to body weight ratio in control and mdx mice at both ages. There was a 22% increase in myosin concentration of mdx DIA at 1 year of age, correlating well with increased normalized active tension in mdx DIA at this age. Also, absolute tetanic tension increased in control and mdx SOL with clenbuterol at both ages. Ventilatory function was significantly impaired in mdx mice at both ages and clenbuterol administration did not alleviate this. Clenbuterol treatment was associated with a 30-40% increase in fatigability in DIA and SOL muscles of control and mdx mice at both ages. Furthermore, 1-year-old mdx mice receiving clenbuterol exhibited deformities in hindlimbs and spine. These results suggest that long-term clenbuterol treatment has a positive effect on muscle growth and force generation, but has adverse side effects such as increased muscle fatigability and development of deformities. PMID- 7477070 TI - Diphtheritic neuropathy. AB - A 29-year-old Haitian man had a sore throat with a marked tonsillar exudate followed by the onset of increasingly blurred vision with swallowing difficulties and, then, a diffuse acute demyelinating neuropathy. Cerebrospinal fluid showed moderate pleiocytosis and elevated protein content. Serological testing for diphtheria, but not for tetanus and poliomyelitis, was markedly positive. Other tests including HIV1 and 2, HTLV1 and Lyme disease were negative. Diphtheritic neuropathy should be considered in the differential diagnosis of Guillain-Barre syndrome in nonvaccinated patients. PMID- 7477071 TI - Spinal motor neuron excitability during the cutaneous silent period. AB - The physiologic mechanisms generating the cutaneous silent period (CSP) remain uncertain. It is not known whether the CSP occurs because of inexcitability of the spinal motor neuron. We therefore, assessed excitability of the motor neuron during the CSP using F-wave responses. H-reflexes were also elicited during the CSP. Electrical stimulation to the fifth digit produced the CSP in the voluntarily contracting abductor pollicis brevis muscle (APB). Median nerve stimulation at the wrist elicited control F or H responses during isometric APB contraction (condition 1) and in resting muscle (condition 2). Control amplitudes were compared to those elicited in the midst of the CSP. In Condition 1, F-wave amplitudes and frequency during the CSP were unchanged compared with controls. However, F-waves were increased in amplitude and frequency during the CSP (P < 0.001) relative to responses elicited in resting muscle (condition 2). H-reflexes during the CSP were suppressed (P < 0.001) compared with controls elicited during contraction (condition 1), but facilitated relative to the resting state (condition 2) in which no H-reflexes were elicitable. We conclude that spinal motor neurons remain excitable to antidromic volleys at the same time that the corticospinal volley is inhibited to produce the CSP. Moreover, motor neuron excitability appears to be increased during the CSP compared to the relaxed state. PMID- 7477072 TI - H-reflexes and F-responses are not equally sensitive to changes in motoneuronal excitability. PMID- 7477073 TI - Thoracoscopic thymectomy in patients with myasthenia gravis. PMID- 7477074 TI - Treatment of Balo's concentric sclerosis with immunosuppressive drugs followed by multimodality evoked potentials and MRI. PMID- 7477076 TI - Reasons for failures in the surgical treatment of thoracic outlet syndrome. PMID- 7477075 TI - Dehydroepiandrosterone sulphate (DHEAS) concentrations and amyotrophic lateral sclerosis. PMID- 7477077 TI - Somatosensory evoked potentials of the phrenic nerve. PMID- 7477078 TI - The clinical correlates of IgG antiganglioside antibodies. PMID- 7477079 TI - Decomposition of the EMG signal into constituent motor unit action potentials. PMID- 7477081 TI - Can proximal nerve lesions give rise to a "Tinel-like" sign distally? PMID- 7477080 TI - Middle-latency responses of the orbicularis oculi muscle: dissimilar origin of the "relaxed" and "contracted" motor evoked potentials. PMID- 7477082 TI - The diagnostic utility of F waves in L5/S1 radiculopathy. PMID- 7477083 TI - Identification of yeasts by hydrolysis of amides. AB - We analysed the hydrolysis of 23 amides by 500 yeast and yeast-like strains isolated from clinical specimens, identified to species level by conventional methods, in order to verify the validity of this method of species identification. The results show that 10 of these amides (acetamide, acrylamide, alaninamide, formamide, glycinamide, propionamide, urea, thioacetamide, thiourea and valeramide) are sufficient to differentiate seven genera and 19 species, with an occasional requirement for three additional tests: cycloheximide susceptibility, surface film formation on liquid medium and ascospore formation. The study of the amidase activity in yeasts and yeast-like fungi seems to be a promising method of identifying strains isolated from clinical samples. PMID- 7477084 TI - Identification of oral yeast species isolated from individuals with diabetes mellitus. AB - In our epidemiological study of 439 patients with diabetes mellitus, the proportion of Candida albicans isolated by the oral rinse technique was 67%. A comparison of the conventional germ tube test with the API 20C Auxanogram kit revealed that 23.6% (129/546) of germ tube-positive species were not identified as C. albicans by the kit. The API 20C Auxanogram therefore underestimated the prevalence of C. albicans. Additionally, a significant number of yeasts (138/1050, 13.1%) isolated from these patients could not be reliably identified by the kit. PMID- 7477085 TI - Origin of differences in susceptibility of Candida krusei to azole antifungal agents. AB - Two Candida krusei isolates were used to compare the effects of fluconazole, ketoconazole and itraconazole on growth and ergosterol synthesis, and to measure intracellular drug contents. Fifty per cent inhibition (IC50) of growth was achieved at 0.05-0.08 microM itraconazole and 0.56-1.2 microM ketoconazole, whereas 91-->100 microM fluconazole was needed to reach the IC50 value. Similar differences in sensitivity to these azole antifungal agents were seen when their effects on ergosterol synthesis from [14C]acetate were measured after 4 h and 24 h of growth. However, when the effects of the azoles on ergosterol synthesis from [14C]mevalonate by subcellular fractions were measured, fluconazole was only 2.3 6.1 times less active than itraconazole, and the IC50 values for ketoconazole were almost similar to those obtained with itraconazole. These results indicate that differences in susceptibility to itraconazole and ketoconazole are unrelated to differences in affinity for the C. krusei cytochrome P450. The much lower growth-inhibitory effects of fluconazole can also be explained partly only by a lower affinity for the P450-dependent 14 alpha-demethylase. The differences in sensitivity of both C. krusei isolates appeared to arise from differences in the intracellular itraconazole, ketoconazole and fluconazole contents. Depending on the experimental conditions, these isolates accumulated 6-41 times more itraconazole than ketoconazole and the intracellular ketoconazole content was 3.0 19.0 times higher than that of fluconazole. PMID- 7477086 TI - Mycoserological study of the treatment of paediatric cystic fibrosis patients with Saccharomyces boulardii (Saccharomyces cerevisiae Hansen CBS 5926). AB - Saccharomyces boulardii (SB) (Saccharomces cerevisiae Hansen CBS 5926) is a yeast widely used in humans for the prevention and treatment of infectious enterocolitis. SB is said also to antagonize Candida albicans when given orally to living organisms. This double-blind trial was performed to determine the effect and tolerance of SB as an oral therapeutic in patients suffering from cystic fibrosis receiving long-term treatment with cephalosporins or cotrimoxazole, by examining C. albicans counts in the intestine. Extensive mycoserological examinations for drug safety evaluation were also performed. To be selected for the study patients had to present C. albicans in their intestinal flora. None of the patients enrolled exhibited clinical symptoms of candidosis. A daily dose of 750 mg (250 mg t.i.d.) of lyophilized SB given for 21 days did not affect the number of C. albicans commensals in those patients. However, the mycoserological data confirmed the safety of SB treatment with respect to a hypothetically possible SB fungaemia and a possible falsification of Candida serology. PMID- 7477087 TI - Chronic meningitis caused by Trichosporon beigelii in India. AB - A 36-year-old woman developed fatal meningitis following a myelogram. Trichosporon beigelii could be grown from the cerebrospinal fluid (CSF) on two occasions. This represents the first culture-proven case of meningitis due to Trichosporon beigelii. PMID- 7477088 TI - Fonsecaea pedrosoi isolated from skin crusts of Bowen's disease. AB - Fonsecaea pedrosoi was isolated from scaly crusts of plaque lesions on the left thigh of a 60-year-old Japanese man. Histological examination confirmed these lesions to be due to Bowen's disease, and sclerotic cells were recognized in the stratum corneum. To our knowledge, this is the first study to demonstrate superficial colonization of a pre-existing skin lesion by F. pedrosoi. PMID- 7477089 TI - Cryptococcosis in a cat seropositive for feline immunodeficiency virus. AB - A 5-year-old female Siamese cat was presented to the veterinary teaching hospital with a history of bronchopneumonia for 20 days. The cat had not responded to antibacterial chemotherapy and had developed a pronounced submandibular lymphadenopathy. Characteristically encapsulated yeast cells with narrow-necked buds were clearly seen in a fine-needle aspirate of the lymph node with an India ink preparation. Cryptococcus neoformans var. neoformans was identified. Susceptibility tests on the isolated strain were performed using antifungal tablets. The strain was sensitive to amphotericin B, fluconazole, itraconazole and ketoconazole and was resistant to 5-fluorocytosine. The cat was positive for feline immunodeficiency virus. Nevertheless, the cat was treated with ketoconazole for 3 months and apparently recovered. Three months later the animal was presented in a precomatose state. The owners refused to treat the animal and the cat was destroyed. PMID- 7477090 TI - Clinical and pharmacokinetic investigations of oral terbinafine in patients with tinea unguium. AB - A clinical trial of once-daily administration of a 125-mg tablet of terbinafine, an oral antimycotic agent, was performed on patients with tinea unguium to evaluate its efficacy, safety, possible side-effects and its incorporation into nails and hair. Thirty-four patients were recruited into the study. For the statistical analysis, one of these patients was used only for the safety rating. Accordingly, 33 patients were used for the efficacy rating, and all 34 patients were employed for the safety rating. The efficacy rating in the overall efficacy evaluation was 90.9% (30/33). No adverse effects, including abnormal changes in laboratory test values, were observed. A pharmacokinetic study revealed that terbinafine was detected in the nail tissue at and after week 2. It reached 0.78 ng mg-1 at the end of week 12 and remained at almost the same level thereafter. Terbinafine was also detected in hair at and after week 23. The average value was 3.14 ng mg-1. The plasma concentration of the drug reached a steady state (280.3 ng ml-1) at approximately week 10, and no tendency to further accumulation was noted. These results confirm the favourable incorporation of terbinafine into nail and hair. On the basis of these results, it was concluded that the drug demonstrates excellent efficacy and satisfactory safety in patients with tinea unguium. The pharmacokinetic investigation also demonstrated its excellent treatment efficacy. PMID- 7477091 TI - Long-term maintenance of lyophilized fungal cultures of the genera Epidermophyton, Microsporum, Paecilomyces and Trichophyton. AB - Twenty-six sporulating strains of lyophilized microscopic fungi of the genera Epidermophyton, Microsporum, Paecilomyces and Trichophyton have been maintained at 2-8 degrees C for 8-14 years. In 23 strains the number of viable particles reduced less than 10-fold during this period compared with immediately after lyophilization. The viability of other three investigated strains ranged from 3.4% to 7.1% in comparison with the original values. PMID- 7477092 TI - Microsporum nanum--a report from Malawi (Africa). AB - Microsporum nanum Fuentes 1956 has been identified in scrapings from a skin lesion in a patient in Karonga District, Malawi, Africa. We believe that this is the first such identification from the African continent. The patient had a single skin lesion in his popliteal fossa and was known to rear pigs. PMID- 7477093 TI - Dermatophytosis in northern Greece during the decade 1981-1990. AB - Dermatophytic infections are very common in Greece. In the Mycological Laboratory of the Venereal and Skin Diseases Hospital in Thessaloniki, 6572 isolates of different dermatophytes were obtained from 17,120 patients examined. It is suggested that 5% of the people who present with skin problems in Greece suffer from dermatophyte infections. They are frequent causative agents of tinea pedis, tinea cruris, tinea corporis, tinea capitis and tinea unguium. In this paper, the species, the number and the prevalence of the dermatophytes were studied according to location and sex of the patients. The contribution of dermatophyte infections to the overall incidence of superficial fungal infection over 10 years was also studied. PMID- 7477094 TI - The occurrence of dermatophytes in Slovakia. AB - We have compared the epidemiology of dermatophytoses in Slovakia during the periods 1956-85 and 1986-92. Infections caused by Trichophyton rubrum gradually increased while infections due to Trichophyton verrucosum and Microsporum canis decreased. PMID- 7477095 TI - Isolation of dermatophytes and other keratinophilic fungi from surface sediments of the Shatt Al-Arab River and its creeks at Basrah, Iraq. AB - Twenty-five sediment samples were taken from randomly selected sites in the Shatt Al-Arab River and its creeks and analysed for dermatophytes and related keratinophilic fungi. The results revealed that out of 25 samples only 13 (52%) yielded dermatophytes and related keratinophilic fungi. A total of nine species in four genera were isolated. The most frequent genera isolated in this study were Chrysosporium and its teleomorph Aphanoascus. The species most frequently found were Aphanoascus fulvescens, A. durus, Chrysosporium crassitunicatum, Chr. keratinophilum and Chr. tropicum (each n = 3). Microsporum was represented by two species, namely M. fulvum and M. gypseum. Trichophyton was represented by one species, T. verrucosum. The occurrence of these fungi illustrates that sediments may act as a reservoir for potentially pathogenic fungi for human and animals. PMID- 7477096 TI - Mycotic pulmonary disease by Beauveria bassiana in a captive tortoise. AB - A case of fatal pulmonary infection in a female tortoise (Tachemys scripta) imported into Spain from Cuba is reported. Necropsy revealed general pulmonary congestion with pleuritis and a large number of yellowish nodules of the granulomatous type, similar to aspergillomata. Histological examination showed some infiltration of round cells, surrounding a small mass of fungal hyphae. Culturing on Sabouraud glucose agar, demonstrated the presence of a fungus whose macroscopic and microscopic characteristics corresponded to those of Beauveria bassiana. PMID- 7477097 TI - Location in the source chromosome of the 180-kb minichromosome of Leishmania major and characterization of the novel junction. AB - The 180-kb LD1 minichromosome of Leishmania major (M180) is a large inverted duplication which arises spontaneously from a megabasic chromosome. In this work this locus has been located in the source chromosome at a telomeric position. Both the novel junction created in the middle of the minichromosome, as well as its counterpart region in the source chromosome were cloned and sequenced. Two inverted repeats, which could give rise to two imperfect stem-loops, and an A + T rich DNA sequence were the only particular features observed in this region. A putative model for the formation of the minichromosome is proposed. PMID- 7477098 TI - Characterization and localization of cathepsin B proteinases expressed by adult Ancylostoma caninum hookworms. AB - The hookworm Ancylostoma caninum induces human eosinophilic enteritis (EE), probably via allergic responses to its secretions. Cysteine and metallo proteinases may be the components of these secretions that elicit hypersensitivity reactions. In order to characterize genes encoding cysteine proteinases (CP) secreted by A. caninum, an adult hookworm cDNA library was constructed and screened with a cloned fragment of a hookworm CP gene. This fragment was obtained using consensus oligonucleotide, CP-gene-specific primers in the polymerase chain reaction. cDNAs encoding two CPs were obtained from the library and sequenced. The first gene, AcCP-1, encoded a cathepsin B-like zymogen CP of 343 amino acids (aa), predicted to be processed in vivo into a mature CP of 255 aa. Closest nucleotide identities were to Haemonchus contortus cysteine protease (61%) and to human cathepsin B (60%). The second gene, AcCP-2, encoded a mature CP of 254 aa, that showed 86% identity to AcCP-1, and 58% and 47% identity to bovine cathepsin B and human cathepsin B, respectively. Rabbit antisera raised against recombinant AcCP-1 reacted with esophageal, amphidial and excretory glands in formalin-fixed, paraffin embedded sections of both male and female adult hookworms, and with an antigen of approx. 40 kDa in Western blot analysis of excretory/secretory products from adult hookworms. Together, these immuno hybridization results strongly suggest that the CP encoded by the AcCP-1 gene is secreted by hookworms. These are the first reported CP genes from hookworms. Proteinases encoded by these genes may be responsible for the CP activity that we have shown previously to be secreted by adult A. caninum. PMID- 7477100 TI - Identification of Eimeria bovis merozoite cDNAs using differential mRNA display. AB - Differences in gene expression between Eimeria bovis sporozoites and first generation merozoites were analyzed using the technique of differential mRNA display. Approx. 5% of the sequences detected in first-generation merozoites appear to be unique relative to sporozoites. Several of the bands corresponding to merozoite-specific gene expression were isolated and cloned. Northern blot analysis revealed that the cDNA fragments DMZ-7, DMZ-8 and NMZ-6 hybridized to mRNAs expressed at > 50-fold higher levels in merozoites relative to sporozoites. A fourth cDNA fragment, NMZ-4, hybridized to a mRNA expressed at 3-fold higher levels in merozoites. Further characterization demonstrated that expression of DMZ-8 in E. bovis-infected bovine cells begins as early as 12 h after sporozoite invasion and continues throughout the entire 14 days of first-generation schizogony. Sequence analysis of each of the four merozoite cDNAs failed to identify any significant similarity to any entries in the GenBank database, suggesting that these developmentally regulated genes may be unique to coccidian parasites. PMID- 7477099 TI - Female-specific sequences isolated from Schistosoma mansoni by representational difference analysis. AB - Representational difference analysis (RDA), a recently described, polymerase chain-reaction-based modification of the subtractive hybridization process was employed to isolate female-specific sequences of Schistosoma mansoni. Using HindIII-derived amplicons, an excess of male schistosome DNA was employed to remove sequences common to both male and female adult S. mansoni genomes from female genomic DNA. Following three rounds of RDA, the enriched sequences included two female-specific repetitive elements. One of these exhibited 76% homology to the SM alpha family of retroposons and represents a W chromosome specific variant of that family. The other sequence represents a novel, female specific repetitive sequence. These sequences have been designated SM alpha fem-1 and W2, respectively, and both are apparently arrayed as tandem repeats on the W chromosome of S. mansoni. The isolation and characterization of the two female specific sequences by RDA indicates that this procedure should also find utility in the definition of traits and sequences that differ among other groups of schistosomes. PMID- 7477101 TI - Kinetics and molecular characteristics of arginine transport by Leishmania donovani promastigotes. AB - Characteristics of transport of L-arginine were studied in Leishmania donovani promastigotes grown in vitro in a defined medium. The promastigotes exhibited a time-dependent, temperature-sensitive, pH-dependent and saturable uptake of arginine. Metabolic inhibitors caused 81-92% inhibition, indicating that arginine influx in promastigotes is an energy requiring process. The presence of Na+ ions was necessary for full activity. Considerable inhibition was also noticed with valinomycin, gramicidin and amiloride. The transporter seems to involve an -SH group at the active site. The most distinctive feature of the leishmanial transporter was that lysine and ornithine did not show significant competition with arginine transport. Other neutral and acidic amino acids, as well as polyamines were also ineffective. The arginine analogues, viz., nitro-L-arginine methyl ester, N-nitro-L-arginine, aminoguanidine, agmatine and D-arginine were not recognised by the transporter, while N-methyl-L-arginine acetate and phospho L-arginine showed competition, indicating stereo-specificity of the transporter and recognition of both the guanidino group, as well as the arginine side chain by the transporter. No exchange of intracellular [14C]arginine taken up by the promastigotes was noticed during incubation with 2 or 5 mM arginine in the extracellular medium. Eighty percent of the arginine taken up remained in the trichloroacetic acid-soluble fraction. Pentamidine caused competitive inhibition of arginine transport, exhibiting an IC50 value of 40 microM. Results indicate the presence of a novel distinct arginine transporter in Leishmania promastigotes. PMID- 7477102 TI - Predicted disulfide-bonded structures for three uniquely related proteins of Plasmodium falciparum, Pfs230, Pfs48/45 and Pf12. AB - Pfs230 is a surface protein of the gametes of Plasmodium falciparum and has been demonstrated to be a target of malaria transmission-blocking antibodies; it is an important candidate antigen for a transmission-blocking vaccine. The target epitopes of transmission-blocking antibodies against Pfs230 are almost all reduction sensitive suggesting that disulfide bonds are critical for folding the native molecule. Following the cloning of the Pfs230 gene attempts are now underway to express subunits of the protein for use in vaccine trials. It will be important to understand the disulfide-bond structure of the Pfs230 to achieve this goal. In this paper we present a model for this structure based on the observation that the Pfs230 molecule contains a series of regularly repeated cysteine-containing motifs. Four such motifs have been identified, together with a fifth cysteineless motif, which occur in the same relative order, with regular alternating omission of specific motifs, 14 times throughout the length of the protein. Each of the 14 sets of motifs contains an even number of cysteine residues (2, 4 or 6). We postulate that each set folds into a separate disulfide bonded domain in which corresponding pairs of cysteines form an equivalent disulfide bond in every such domain. The postulated bonding arrangements in the different domains are mutually confirmatory throughout the sequence of Pfs230. We have identified two other malaria proteins, Pfs48/45 and Pf12, which share the same arrangements of motifs and conform to the same disulfide-bond structure proposed for Pfs230; no other proteins in the sequence data base share these characteristics.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7477103 TI - Differential susceptibility of filarial and human erythrocyte glutathione reductase to inhibition by the trivalent organic arsenical melarsen oxide. AB - The glutathione reductases (GR) from two cattle filariae (Setaria digitata and Onchocerca gutturosa) have been isolated and their properties have been compared to those of human erythrocyte GR. In general, the enzymes appear to be very similar with respect to substrate-specificity for glutathione disulfide and NADPH, molecular mass (97 kDa vs. 98 kDa) and oligomeric organisation (subunit size of 51 kDa vs. 50 kDa). However, studies on the inhibition of the enzymes by the trivalent melaminophenyl arsenical melarsen oxide revealed that the human GR is less susceptible to inhibition by the arsenical than the filarial enzymes. Further, it was found that the mechanism of inactivation differs for the host and filarial enzymes. The human enzyme is inhibited by melarsen oxide in a competitive manner with a Ki of 23.7 microM, whereas the filarial GRs are inhibited in two stages: an immediate partial inactivation followed by a time dependent stage with saturable pseudo-first-order kinetics. Ki values for the S. digitata and O. gutturosa GRs are 38.3 microM and 4.5 microM, respectively, with maximum second-stage inactivation rates of 1.0 x 10(-4) s-1 and 24.3 x 10(-4) s 1, respectively. These differences between host and parasite enzyme might reflect differences in the primary and secondary structure of the proteins which might be exploitable for the design of new specific macrofilaricidal drugs. PMID- 7477104 TI - Cloning of the gene for phosphoglycerate kinase from Schistosoma mansoni and characterization of its gene product. AB - As molecules on the surface or associated with the outer covering (tegument) of Schistosoma mansoni are a major focus as potential vaccine candidates, affinity purified antibodies which are specific to the tegumental antigens were used to immunoscreen a lambda gt11 S. mansoni cercarial cDNA library. One of the identified clones was found to encode the glycolytic enzyme phosphoglycerate kinase (PGK, EC 2.7.2.3). The 1.5-kb cDNA clone has a single open reading frame encoding 416 amino acids and exhibits over 60% identity to PGKs from a number of eukaryotic species. Recombinant S. mansoni PGK (SmPGK) was overexpressed in Escherichia coli, purified, and shown to have PGK enzyme activity. Native protein affinity purified from S. mansoni adult worms was shown by microsequencing to have the same amino-acid sequence as deduced from the cDNA sequence, thus confirming the cDNA clone we identified encodes S. mansoni phosphoglycerate kinase. Antibodies localize the native SmPGK to various tissues including the tegument of 3-h schistosomula and 42-day adult worms. PMID- 7477105 TI - Comparison of introns in a cdc2-homologous gene within a number of Plasmodium species. AB - The first three introns of CRK2, a cdc2-homologous gene, have been compared in a total of seven Plasmodium species. The introns were located at conserved sites, suggesting an ancestral origin. Interspecific comparison of intron sequences agreed with the previously inferred evolutionary relationships of the malaria parasites. Unlike the introns in the rodent malaria species, the similarity of the CRK2 introns was regionalized between the human parasite P. vivax and the simian parasite P. knowlesi: the central region of all three introns showed markedly less interspecific similarity than the 5' and 3' regions. This was also in contrast with the organisation and composition of homologous intron pairs from other genes of the same two species. No conservation at the level of secondary structure could be detected, even between highly similar introns. A database search for intron-containing Plasmodium genes was performed. All introns obtained in this way plus the additional CRK2 introns were scanned for the presence of putative branching site consensus sequences. For P. falciparum, we present an update of the 5' and 3' splice-site consensus. PMID- 7477106 TI - Sequence and organization of an unusual histone H4 gene in the human parasite Entamoeba histolytica. PMID- 7477107 TI - A Plasmodium falciparum gene encoding a high mobility group protein box. PMID- 7477108 TI - Genus- and subgenus-specific oligonucleotide probes for Acanthamoeba. PMID- 7477109 TI - Cloning and characterization of the ribosomal l11 gene from Leishmania spp. PMID- 7477110 TI - Stable episomal transfection of Entamoeba histolytica. PMID- 7477111 TI - The female-specific W1 sequence of the Puerto Rican strain of Schistosoma mansoni occurs in both genders of a Liberian strain. PMID- 7477112 TI - Molecular karyotype of clone CL Brener chosen for the Trypanosoma cruzi genome project. PMID- 7477113 TI - Primary structure of a putative adenylate kinase gene of Giardia lamblia. PMID- 7477114 TI - Stage-specifically expressed genes of Angiostrongylus cantonensis: identification by differential display. PMID- 7477115 TI - Sequence variations in the non-repetitive regions of the liver stage-specific antigen-1 (LSA-1) of Plasmodium falciparum from field isolates. PMID- 7477116 TI - Teratogenicity of high vitamin A intake. AB - BACKGROUND: Studies in animals indicate that natural forms of vitamin A are teratogenic. Synthetic retinoids chemically similar to vitamin A cause birth defects in humans; as in animals, the defects appear to affect tissues derived from the cranial neural crest. METHODS: Between October 1984 and June 1987, we identified 22,748 pregnant women when they underwent screening either by measurement of maternal serum alpha-fetoprotein or by amniocentesis. Nurse interviewers obtained information on the women's diet, medications, and illnesses during the first trimester of pregnancy, as well as information on their family and medical history and exposure to environmental agents. We obtained information on the outcomes of pregnancy from the obstetricians who delivered the babies or from the women themselves. Of the 22,748 women, 339 had babies with birth defects; 121 of these babies had defects occurring in sites that originated in the cranial neural crest. RESULTS: For defects associated with cranial-neural crest tissue, the ratio of the prevalence among the babies born to women who consumed more than 15,000 IU of preformed vitamin A per day from food and supplements to the prevalence among the babies whose mothers consumed 5000 IU or less per day was 3.5 (95 percent confidence interval, 1.7 to 7.3). For vitamin A from supplements alone, the ratio of the prevalence among the babies born to women who consumed more than 10,000 IU per day to that among the babies whose mothers consumed 5000 IU or less per day was 4.8 (95 percent confidence interval, 2.2 to 10.5). Using a smoothed regression curve, we found an apparent threshold near 10,000 IU per day of supplemental vitamin A. The increased frequency of defects was concentrated among the babies born to women who had consumed high levels of vitamin A before the seventh week of gestation. CONCLUSIONS: High dietary intake of preformed vitamin A appears to be teratogenic. Among the babies born to women who took more than 10,000 IU of preformed vitamin A per day in the form of supplements, we estimate that about 1 infant in 57 had a malformation attributable to the supplement. PMID- 7477117 TI - Campylobacter jejuni infection and Guillain-Barre syndrome. AB - BACKGROUND: Although infection with Campylobacter jejuni is recognized as a common antecedent of the Guillain-Barre syndrome, the clinical and epidemiologic features of this association are not well understood. METHODS: We performed a prospective case-control study in a cohort of patients with Guillain-Barre syndrome (96 patients) or Miller Fisher syndrome (7 patients) who were admitted to hospitals throughout England and Wales between November 1992 and April 1994. Bacteriologic and serologic techniques were used to diagnose preceding C. jejuni infection. RESULTS: There was evidence of recent C. jejuni infection in 26 percent of the patients with Guillain-Barre or Miller Fisher syndrome, as compared with 2 percent of household controls and 1 percent of age-matched hospital controls (P < 0.001). Of the 27 patients with C. jejuni infection, 19 (70 percent) reported having had a diarrheal illness within 12 weeks before the onset of the neurologic illness. No specific serotypes were associated with Guillain-Barre syndrome. C. jejuni infection was slightly more common in men (P = 0.14) and was more likely to be associated with a pure motor syndrome and a slower recovery (P = 0.03). The patients with preceding C. jejuni infection were more likely to have acute axonal neuropathy or axonal degeneration in association with acute inflammatory demyelinating polyradiculoneuropathy, and they had greater disability after one year (P = 0.02). C. jejuni infection was significantly associated with a poor outcome even after correction for other factors associated with a poor prognosis. CONCLUSIONS: Infection with C. jejuni often precedes the Guillain-Barre syndrome and is associated with axonal degeneration, slow recovery, and severe residual disability. PMID- 7477118 TI - Expression of P-glycoprotein in high-grade osteosarcomas in relation to clinical outcome. AB - BACKGROUND: Increased levels of P-glycoprotein occur in some osteosarcomas. In this study we determined the relation between P-glycoprotein status and outcome in patients with high-grade osteosarcoma. METHODS: P-glycoprotein status was determined immunohistochemically in specimens of osteosarcoma of the extremities (stage II) from 92 patients who were treated with surgery and chemotherapy. The P glycoprotein status was analyzed in relation to the length of event-free survival. RESULTS: The presence of increased levels of P-glycoprotein in the osteosarcoma was significantly associated with a decreased probability of remaining event-free after diagnosis (P = 0.002). In a multivariate analysis, P glycoprotein status (P = 0.001) and the extent of tumor necrosis after preoperative chemotherapy (P = 0.04) were independent predictors of clinical outcome. The risk of adverse events was increased substantially (rate ratio, 3.37; 95 percent confidence interval, 1.60 to 7.10) among patients with increased levels of P-glycoprotein in tumor cells, as compared with patients who did not have increased levels of P-glycoprotein in tumor cells. CONCLUSIONS: In patients with high-grade osteosarcoma treated with surgery and chemotherapy, the presence of increased levels of P-glycoprotein in tumor cells is associated with a significantly increased risk of adverse events and is independent of the extent of necrosis after preoperative chemotherapy. PMID- 7477119 TI - Brief report: impaired processing of prohormones associated with abnormalities of glucose homeostasis and adrenal function. PMID- 7477120 TI - Images in clinical medicine. Kawasaki's disease. PMID- 7477121 TI - Primary prevention of stroke. PMID- 7477123 TI - Case records of the Massachusetts General Hospital. Weekly clinicopathological exercises. Case 36-1995. A 61-year-old man with increasing weakness and atrophy of all extremities. PMID- 7477122 TI - Screening for prostate cancer with prostate-specific antigen. An examination of the evidence. PMID- 7477124 TI - Vitamin A and birth defects. Continuing caution is needed. PMID- 7477125 TI - The changing concepts of Guillain-Barre syndrome. PMID- 7477126 TI - P-glycoprotein--a marker of cancer-cell behavior. PMID- 7477127 TI - The computer-based patient record and confidentiality. PMID- 7477128 TI - Breast implants and connective-tissue diseases. PMID- 7477129 TI - Breast implants and connective-tissue diseases. PMID- 7477131 TI - Clinical problem-solving: diverted by the chief complaint. PMID- 7477130 TI - Breast implants and connective-tissue diseases. PMID- 7477132 TI - Clinical problem-solving: diverted by the chief complaint. PMID- 7477133 TI - Clinical problem-solving: diverted by the chief complaint. PMID- 7477134 TI - Clinical problem-solving: diverted by the chief complaint. PMID- 7477135 TI - Clinical problem-solving: diverted by the chief complaint. PMID- 7477136 TI - Clinical Problem-Solving: diverted by the chief complaint. PMID- 7477137 TI - Gastric carcinoma. PMID- 7477138 TI - Gastric carcinoma. PMID- 7477139 TI - Gastric carcinoma. PMID- 7477141 TI - Diltiazem and subacute cutaneous lupus erythematosus-like lesions. PMID- 7477140 TI - Renal osteodystrophy. PMID- 7477142 TI - Persistence of differences in iodine status in newborns after the reunification of Berlin. PMID- 7477143 TI - Effect of oral alendronate on bone mineral density and the incidence of fractures in postmenopausal osteoporosis. The Alendronate Phase III Osteoporosis Treatment Study Group. AB - BACKGROUND: Postmenopausal osteoporosis is a serious health problem, and additional treatments are needed. METHODS: We studied the effects of oral alendronate, an aminobisphosphonate, on bone mineral density and the incidence of fractures and height loss in 994 women with postmenopausal osteoporosis. The women were treated with placebo or alendronate (5 or 10 mg daily for three years, or 20 mg for two years followed by 5 mg for one year); all the women received 500 mg of calcium daily. Bone mineral density was measured by dual-energy x-ray absorptiometry. The occurrence of new vertebral fractures and the progression of vertebral deformities were determined by an analysis of digitized radiographs, and loss of height was determined by sequential height measurements. RESULTS: The women receiving alendronate had significant, progressive increases in bone mineral density at all skeletal sites, whereas those receiving placebo had decreases in bone mineral density. At three years, the mean (+/- SE) differences in bone mineral density between the women receiving 10 mg of alendronate daily and those receiving placebo were 8.8 +/- 0.4 percent in the spine, 5.9 +/- 0.5 percent in the femoral neck, 7.8 +/- 0.6 percent in the trochanter, and 2.5 +/- 0.3 percent in the total body (P < 0.001 for all comparisons). The 5-mg dose was less effective than the 10-mg dose, and the regimen of 20 mg followed by 5 mg was similar in efficacy to the 10-mg dose. Overall, treatment with alendronate was associated with a 48 percent reduction in the proportion of women with new vertebral fractures (3.2 percent, vs. 6.2 percent in the placebo group; P = 0.03), a decreased progression of vertebral deformities (33 percent, vs. 41 percent in the placebo group; P = 0.028), and a reduced loss of height (P = 0.005) and was well tolerated. CONCLUSIONS: Daily treatment with alendronate progressively increases the bone mass in the spine, hip, and total body and reduces the incidence of vertebral fractures, the progression of vertebral deformities, and height loss in postmenopausal women with osteoporosis. PMID- 7477144 TI - Effects of radiotherapy and surgery in early breast cancer. An overview of the randomized trials. AB - BACKGROUND: Randomized trials of radiotherapy and surgery for early breast cancer may have been too small to detect differences in long-term survival and recurrence reliably. We therefore performed a systematic overview (meta-analysis) of the results of such trials. METHODS: Information was sought on each subject from investigators who conducted trials that began before 1985 and that compared local therapies for early breast cancer. Data on mortality were available from 36 trials comparing radiotherapy plus surgery with the same type of surgery alone, 10 comparing more extensive surgery with less extensive surgery, and 18 comparing more extensive surgery with less extensive surgery plus radiotherapy. Information on mortality was available for 28,405 women (97.4 percent of the 29,175 women in the trials). RESULTS: The addition of radiotherapy to surgery resulted in a rate of local recurrence that was three times lower than the rate with surgery alone, but there was no significant difference in 10-year survival; among a total of 17,273 women enrolled in such trials, mortality was 40.3 percent with radiotherapy and 41.4 percent without radiotherapy (P = 0.3). Radiotherapy was associated with a reduced risk of death due to breast cancer (odds ratio, 0.94; 95 percent confidence interval, 0.88 to 1.00; P = 0.03), which indicates that, after 10 years, there would be about 0 to 5 fewer deaths due to breast cancer per 100 women. However, there was an increased risk of death from other causes (odds ratio, 1.24; 95 percent confidence interval, 1.09 to 1.42; P = 0.002). This, together with the age-specific death rates, implies, after 10 years, a few extra deaths not due to breast cancer per 100 older women or per 1000 younger women. During the first decade or two after diagnosis, the excess in the rate of such deaths that was associated with radiotherapy was much greater women who were over 60 years of age at randomization (15.3 percent vs. 11.1 percent [339 vs. 249 deaths]) than among those under 50 (2.5 percent vs. 2.0 percent [62 vs. 49 deaths]). Breast-conserving surgery involved some risk of recurrence in the remaining tissue, but no significant differences in overall survival at 10 years were found in the studies of mastectomy versus breast-conserving surgery plus radiotherapy (4891 women), more extensive surgery versus less extensive surgery (4818 women), or axillary clearance versus radiotherapy as adjuncts to mastectomy (4370 women). CONCLUSIONS: Some of the local therapies for breast cancer had substantially different effects on the rates of local recurrence--such as the reduced recurrence with the addition of radiotherapy to surgery--but there were no definite differences in overall survival at 10 years. PMID- 7477145 TI - Reanalysis and results after 12 years of follow-up in a randomized clinical trial comparing total mastectomy with lumpectomy with or without irradiation in the treatment of breast cancer. AB - BACKGROUND: Previous findings from a clinical trial (Protocol B-06) conducted by the National Surgical Adjuvant Breast and Bowel Project (NSABP) indicated the worth of lumpectomy and breast irradiation for treating breast cancer. After the discovery by NSABP staff members of falsified information on patients enrolled in the study by St. Luc Hospital in Montreal, separate audits were conducted at St. Luc Hospital and other participating institutions. We report the results of both audits and update the study findings through an average of 12 years of follow-up. METHODS: Patients with either negative or positive axillary nodes and tumors 4 cm or less in diameter were randomly assigned to one of three treatments: total mastectomy, lumpectomy followed by breast irradiation, or lumpectomy without irradiation. Three cohorts of patients were analyzed. The first cohort included all 2105 randomized patients, who were analyzed according to the intention-to treat principle. The second cohort consisted of 1851 eligible patients in the first cohort with known nodal status who agreed to be followed and who accepted their assigned therapy (among those excluded were 6 patients from St. Luc Hospital who were declared ineligible because of falsified biopsy dates). The third cohort consisted of the patients in the second cohort minus the 322 eligible patients from St. Luc Hospital (total, 1529 patients). RESULTS: Regardless of the cohort, no significant differences were found in overall survival, disease-free survival, or survival free of disease at distant sites between the patients who underwent total mastectomy and those treated by lumpectomy alone or by lumpectomy plus breast irradiation. After 12 years of follow-up, the cumulative incidence of a recurrence of tumor in the ipsilateral breast was 35 percent in the group treated with lumpectomy alone and 10 percent in the group treated with lumpectomy and breast irradiation (P < 0.001). CONCLUSIONS: Our findings continue to indicate that lumpectomy followed by breast irradiation is appropriate therapy for women with either negative or positive axillary nodes and breast tumors 4 cm or less in diameter. PMID- 7477146 TI - Effects of regular exercise on blood pressure and left ventricular hypertrophy in African-American men with severe hypertension. AB - BACKGROUND: The prevalence of hypertension and its cardiovascular complications is higher in African Americans than in whites. Interventions to control blood pressure in this population are particularly important. Regular exercise lowers blood pressure in patients with mild-to-moderate hypertension, but its effects in patients with severe hypertension have not been studied. We examined the effects of moderately intense exercise on blood pressure and left ventricular hypertrophy in African-American men with severe hypertension. METHODS: We randomly assigned 46 men 35 to 76 years of age to exercise plus antihypertensive medication (23 men) or antihypertensive medication alone (23 men). A total of 18 men in the exercise group completed 16 weeks of exercise, and 14 completed 32 weeks of exercise, which was performed three times per week at 60 to 80 percent of the maximal heart rate. RESULTS: After 16 weeks, mean (+/- SD) diastolic blood pressure had decreased from 88 +/- 7 to 83 +/- 8 mm Hg in the patients who exercised, whereas it had increased slightly, from 88 +/- 6 to 90 +/- 7 mm Hg, in those who did not exercise (P = 0.002). Diastolic blood pressure remained significantly lower after 32 weeks of exercise, even with substantial reductions in the dose of antihypertensive medication. In addition, the thickness of the interventricular septum (P = 0.03), the left ventricular mass (P = 0.02), and the mass index (P = 0.04) had decreased significantly after 16 weeks in the patients who exercised, whereas there was no significant change in the nonexercisers. CONCLUSIONS: Regular exercise reduced blood pressure and left ventricular hypertrophy in African-American men with severe hypertension. PMID- 7477147 TI - Images in clinical medicine. Osteoporosis in a woman of the early Bronze Age. PMID- 7477148 TI - The National Cancer Institute audit of the National Surgical Adjuvant Breast and Bowel Project Protocol B-06. AB - BACKGROUND: The National Surgical Adjuvant Breast and Bowel Project (NSABP) Protocol B-06, a clinical trial sponsored by the National Cancer Institute (NCI), has provided evidence of the value of lumpectomy and breast irradiation for treating women with breast cancer in an early stage. Publicity generated by the discovery that the study included fraudulent data on patients enrolled by St. Luc Hospital in Montreal aroused concern about the overall accuracy of the data and conclusions. To address this concern, the NCI conducted an audit of other participating institutions. METHODS: In 1994, data on 1554 of the 1809 randomized patients (85.9 percent) enrolled by centers other than St. Luc Hospital were audited at 37 clinical sites in North America. The audit included data on eligibility, survival, disease-free survival, the length of time to a recurrence of cancer in the ipsilateral breast, and documentation of signed informed consent. RESULTS: End points were assessed for all 1554 patients, and eligibility was assessed for 1507 patients; 47 patients were excluded because their forms were not complete or not returned. A total of 1429 patients had their eligibility status verified. Of a total of 7770 data points examined with respect to the number of positive nodes at base line, treatment characteristics, first events (excluding death), recurrence of cancer in the ipsilateral breast, and survival, 7577 (97.5 percent) were verified, 123 (1.6 percent) could not be verified, and 70 (0.9 percent) were discrepant with the NSABP file. Of the 1554 patients, 1340 (86.2 percent) had all audited items (including eligibility) verified, 69 (4.4 percent) had at least one discrepant item, and 113 (7.3 percent) had at least one unverified item (as a result of missing or incomplete data); 32 (2.1 percent) were not assessed for eligibility but had no other discrepant or unverifiable items. Written informed consent was documented for 1098 patients before surgery and 210 after surgery; no date appeared on the signed form for 137. The informed consent status was not verified for 71 patients and could not be determined for 38. The rates of verification of end-point data and documentation of written informed consent were similar among the total-mastectomy group, the lumpectomy group, and the group treated by lumpectomy and breast irradiation. CONCLUSIONS: The audit confirms the adequacy of the data on which the reanalysis of Protocol B 06 and the results after 12 years of follow-up are based. PMID- 7477149 TI - Management of acquired bullous skin diseases. PMID- 7477151 TI - The treatment of postmenopausal osteoporosis. PMID- 7477150 TI - Case records of the Massachusetts General Hospital. Weekly clinicopathological exercises. Case 37-1995. A 6-year-old boy with a rash, meningismus, and diplegia. PMID- 7477152 TI - Surgery for early breast cancer--can less be more? PMID- 7477153 TI - Xenotransplantation and xenogeneic infections. PMID- 7477154 TI - Attenuation of isoproterenol-mediated vasodilatation in blacks. PMID- 7477155 TI - Attenuation of isoproterenol-mediated vasodilatation in blacks. PMID- 7477156 TI - Attenuation of isoproterenol-mediated vasodilatation in blacks. PMID- 7477157 TI - Optimal oral anticoagulation for patients with nonrheumatic atrial fibrillation and recent cerebral ischemia. PMID- 7477158 TI - Optimal oral anticoagulation for patients with mechanical heart valves. PMID- 7477159 TI - Clarithromycin-related toxic effects of digoxin. PMID- 7477160 TI - Propofol and postoperative infections. PMID- 7477161 TI - Propofol and postoperative infections. PMID- 7477162 TI - Propofol and postoperative infections. PMID- 7477163 TI - More on p53 antigen loss in stored paraffin slides. PMID- 7477164 TI - More on extended-release acetaminophen. PMID- 7477165 TI - Timing of sexual intercourse in relation to ovulation. Effects on the probability of conception, survival of the pregnancy, and sex of the baby. AB - BACKGROUND: The timing of sexual intercourse in relation to ovulation strongly influences the chance of conception, although the actual number of fertile days in a woman's menstrual cycle is uncertain. The timing of intercourse may also be associated with the sex of the baby. METHODS: We recruited 221 healthy women who were planning to become pregnant. At the same time the women stopped using birth control methods, they began collecting daily urine specimens and keeping daily records of whether they had sexual intercourse. We measured estrogen and progesterone metabolites in urine to estimate the day of ovulation. RESULTS: In a total of 625 menstrual cycles for which the dates of ovulation could be estimated, 192 pregnancies were initiated, as indicated by increases in the urinary concentration of human chorionic gonadotropin around the expected time of implantation. Two thirds (n = 129) ended in live births. Conception occurred only when intercourse took place during a six-day period that ended on the estimated day of ovulation. The probability of conception ranged from 0.10 when intercourse occurred five days before ovulation to 0.33 when it occurred on the day of ovulation itself. There was no evident relation between the age of sperm and the viability of the conceptus, although only 6 percent of the pregnancies could be firmly attributed to sperm that were three or more days old. Cycles producing male and female babies had similar patterns of intercourse in relation to ovulation. CONCLUSIONS: Among healthy women trying to conceive, nearly all pregnancies can be attributed to intercourse during a six-day period ending on the day of ovulation. For practical purposes, the timing of sexual intercourse in relation to ovulation has no influence on the sex of the baby. PMID- 7477166 TI - Thyroid hormone treatment after coronary-artery bypass surgery. AB - BACKGROUND: Thyroid hormone has many effects on the cardiovascular system. During and after cardiopulmonary bypass, serum triiodothyronine concentrations decline transiently, which may contribute to postoperative hemodynamic dysfunction. We investigated whether the perioperative administration of triiodothyronine (liothyronine sodium) enhances cardiovascular performance in high-risk patients undergoing coronary-artery bypass surgery. METHODS: We administered triiodothyronine or placebo to 142 patients with coronary artery disease and depressed left ventricular function. The hormone was administered as an intravenous bolus of 0.8 microgram per kilogram of body weight when the aortic cross-clamp was removed after the completion of bypass surgery and then as an infusion of 0.113 microgram per kilogram per hour for six hours. Clinical and hemodynamic responses were serially recorded, as was any need for inotropic or vasodilator drugs. RESULTS: The patients' preoperative serum triiodothyronine concentrations were normal (mean [+/- SD] value, 81 +/- 22 ng per deciliter [1.2 +/- 0.3 nmol per liter]), and they decreased by 40 percent (P < 0.001) 30 minutes after the onset of cardiopulmonary bypass. The concentrations in patients given intravenous triiodothyronine became supranormal and were significantly higher than those in patients given placebo (P < 0.001). However, the concentrations were once again similar in the two groups 24 hours after surgery. The mean postoperative cardiac index was higher in the triiodothyronine group (2.97 +/- 0.72 vs. 2.67 +/- 0.61 liters per minute per square meter of body-surface area, P = 0.007), and systemic vascular resistance was lower (1073 +/- 314 vs. 1235 +/- 387 dyn.sec.cm-5, P = 0.003). The two groups did not differ significantly in the incidence of arrhythmia or the need for therapy with inotropic and vasodilator drugs during the 24 hours after surgery, or in perioperative mortality and morbidity. CONCLUSIONS: Raising serum triiodothyronine concentrations in patients undergoing coronary-artery bypass surgery increases cardiac output and lowers systemic vascular resistance, but does not change outcome or alter the need for standard postoperative therapy. PMID- 7477167 TI - A short-term study of the safety, pharmacokinetics, and efficacy of ritonavir, an inhibitor of HIV-1 protease. European-Australian Collaborative Ritonavir Study Group. AB - BACKGROUND: Reverse-transcriptase inhibitors have only moderate clinical efficacy against the human immunodeficiency virus type 1 (HIV-1). Ritonavir is an inhibitor of HIV-1 protease with potent in vitro anti-HIV properties and good oral bioavailability. METHODS: We evaluated the antiviral activity and safety of ritonavir in a double-blind, randomized, placebo-controlled phase 1 and 2 study of 84 HIV-positive patients with 50 or more CD4+ lymphocytes per cubic millimeter. The patients were randomly assigned to one of four regimens of ritonavir therapy, or to placebo for four weeks and then (by random assignment) to one of the ritonavir regimens. RESULTS: During the first 4 weeks, increases in CD4+ lymphocyte counts and reductions in the log number of copies of HIV-1 RNA per milliliter of plasma were similar among the four dosage groups, but in the three lower-dosage groups there was a return to base-line levels by 16 weeks. After 32 weeks, in the seven patients in the highest-dosage group (600 mg of ritonavir every 12 hours), the median increase from base line in the CD4+ lymphocyte count was 230 cells per cubic millimeter, and the mean decrease in the plasma concentration of HIV-1 RNA (as measured by a branched-DNA assay) was 0.81 log (95 percent confidence interval, 0.40 to 1.22). In a subgroup of 17 patients in the two higher-dosage groups, RNA was also measured with an assay based on the polymerase chain reaction, and after eight weeks of treatment there was a mean maximal decrease in viral RNA of 1.94 log (95 percent confidence interval, 1.37 to 2.51). Adverse events included nausea, circumoral paresthesia, elevated hepatic aminotransferase levels, and elevated triglyceride levels. Ten withdrawals from the study were judged to be related to ritonavir treatment. CONCLUSIONS: In this short-term study, ritonavir was well tolerated and had potent activity against HIV-1, but its clinical benefits remain to be established. PMID- 7477168 TI - A preliminary study of ritonavir, an inhibitor of HIV-1 protease, to treat HIV-1 infection. AB - BACKGROUND: Ritonavir is a potent inhibitor in vitro of human immunodeficiency virus type 1 (HIV-1) protease, which is needed for virions to mature and become infective. We assessed the safety and efficacy of ritonavir in patients with HIV 1 infection. METHODS: We administered ritonavir orally to 62 patients in one of four dosages during a 12-week trial containing a 4-week randomized, placebo controlled, double-blinded phase followed by an 8-week dose-blinded phase. We assessed the response with serial measurements of plasma viremia and serial CD4 cell counts. RESULTS: Fifty-two patients completed the 12-week trial. Diarrhea and nausea were the most common side effects, and reversible elevations in serum triglyceride and gamma-glutamyltransferase levels were the most frequent laboratory abnormalities. Ritonavir had a rapid antiviral effect, with a mean maximal reduction in the number of copies of HIV-1 RNA per milliliter of plasma that ranged from 0.86 to 1.18 log in the four dosage groups. After 12 weeks of treatment, the antiviral effect was partially maintained, with a mean reduction in plasma viremia of 0.5 log. When we used a more sensitive assay for HIV-1 RNA in a subgroup of 20 patients, we found that plasma viremia decreased by a mean of 1.7 log. This antiviral effect was partly sustained at week 12, with a mean reduction of approximately 1.1 log. The patients' CD4 cell counts rose during treatment with ritonavir (median increase, 74 and 83 cells per cubic millimeter at weeks 4 and 12, respectively). CONCLUSIONS: The protease inhibitor ritonavir is well tolerated and has a potent antiviral effect, as shown by substantial decreases in plasma viremia and significant elevations in CD4 cell counts. Expanded clinical trials of ritonavir are warranted. PMID- 7477170 TI - Images in clinical medicine. Macrophages. PMID- 7477169 TI - Autologous bone marrow transplantation as compared with salvage chemotherapy in relapses of chemotherapy-sensitive non-Hodgkin's lymphoma. AB - BACKGROUND: High-dose chemotherapy followed by autologous bone marrow transplantation is a therapeutic option for patients with chemotherapy-sensitive non-Hodgkin's lymphoma who have relapses. In this report we describe a prospective randomized study of such treatment. METHOD: A total of 215 patients with relapses of non-Hodgkin's lymphoma were treated between July 1987 and June 1994. All patients received two courses of conventional chemotherapy. The 109 patients who had a response to chemotherapy were randomly assigned to receive four courses of chemotherapy plus radiotherapy (54 patients) or radiotherapy plus intensive chemotherapy and autologous bone marrow transplantation (55 patients). RESULTS: The overall rate of response to conventional chemotherapy was 58 percent; among patients with relapses after chemotherapy, the response rate was 64 percent, and among those with relapses during chemotherapy, the response rate was 21 percent. There were three deaths from toxic effects among the patients in the transplantation group, and none among those in the group receiving chemotherapy without transplantation. The two groups did not differ in terms of prognostic factors. The median follow-up time was 63 months. The response rate was 84 percent after bone marrow transplantation and 44 percent after chemotherapy without transplantation. At five years, the rate of event-free survival was 46 percent in the transplantation group and 12 percent in the group receiving chemotherapy without transplantation (P = 0.001), and the rate of overall survival was 53 and 32 percent, respectively (P = 0.038). CONCLUSIONS: As compared with conventional chemotherapy, treatment with high-dose chemotherapy and autologous bone marrow transplantation increases event-free and overall survival in patients with chemotherapy-sensitive non-Hodgkin's lymphoma in relapse. PMID- 7477172 TI - Muscle-mediated gene therapy. PMID- 7477171 TI - Pathophysiology of dyspnea. PMID- 7477173 TI - Clinical problem-solving. If at first you don't succeed. PMID- 7477174 TI - Altered thyroid function in nonthyroidal illness and surgery. To treat or not to treat? PMID- 7477175 TI - Pregnancy and the timing of intercourse. PMID- 7477176 TI - Treatment for non-Hodgkin's lymphoma in relapse. What are the alternatives? PMID- 7477177 TI - Cyclosporine and methotrexate for severe rheumatoid arthritis. PMID- 7477178 TI - Cyclosporine and methotrexate for severe rheumatoid arthritis. PMID- 7477179 TI - Cyclosporine and methotrexate for severe rheumatoid arthritis. PMID- 7477180 TI - Cyclosporine and methotrexate for severe rheumatoid arthritis. PMID- 7477181 TI - Cyclosporine and methotrexate for severe rheumatoid arthritis. PMID- 7477182 TI - Cyclosporine and methotrexate for severe rheumatoid arthritis. PMID- 7477183 TI - Glucocorticoids and joint destruction in rheumatoid arthritis. PMID- 7477184 TI - Glucocorticoids and joint destruction in rheumatoid arthritis. PMID- 7477185 TI - Glucocorticoids and joint destruction in rheumatoid arthritis. PMID- 7477186 TI - The prevention of coronary heart disease in women. PMID- 7477187 TI - The prevention of coronary heart disease in women. PMID- 7477188 TI - The prevention of coronary heart disease in women. PMID- 7477189 TI - Angioedema induced by the angiotensin II blocker losartan. PMID- 7477190 TI - Images in clinical medicine: echinococcal cyst of the pericardium. PMID- 7477191 TI - The human molecular genetics network. PMID- 7477192 TI - Tissue plasminogen activator for acute ischemic stroke. AB - BACKGROUND: Thrombolytic therapy for acute ischemic stroke has been approached cautiously because there were high rates of intracerebral hemorrhage in early clinical trials. We performed a randomized, double-blind trial of intravenous recombinant tissue plasminogen activator (t-PA) for ischemic stroke after recent pilot studies suggested that t-PA was beneficial when treatment was begun within three hours of the onset of stroke. METHODS: The trial had two parts. Part 1 (in which 291 patients were enrolled) tested whether t-PA had clinical activity, as indicated by an improvement of 4 points over base-line values in the score of the National Institutes of Health stroke scale (NIHSS) or the resolution of the neurologic deficit within 24 hours of the onset of stroke. Part 2 (in which 333 patients were enrolled) used a global test statistic to assess clinical outcome at three months, according to scores on the Barthel index, modified Rankin scale, Glasgow outcome scale, and NIHSS: RESULTS: In part 1, there was no significant difference between the group given t-PA and that given placebo in the percentages of patients with neurologic improvement at 24 hours, although a benefit was observed for the t-PA group at three months for all four outcome measures. In part 2, the long-term clinical benefit of t-PA predicted by the results of part 1 was confirmed (global odds ratio for a favorable outcome, 1.7; 95 percent confidence interval, 1.2 to 2.6). As compared with patients given placebo, patients treated with t-PA were at least 30 percent more likely to have minimal or no disability at three months on the assessment scales. Symptomatic intracerebral hemorrhage within 36 hours after the onset of stroke occurred in 6.4 percent of patients given t-PA but only 0.6 percent of patients given placebo (P < 0.001). Mortality at three months was 17 percent in the t-PA group and 21 percent in the placebo group (P = 0.30). CONCLUSIONS: Despite an increased incidence of symptomatic intracerebral hemorrhage, treatment with intravenous t PA within three hours of the onset of ischemic stroke improved clinical outcome at three months. PMID- 7477193 TI - Low-molecular-weight heparin for the treatment of acute ischemic stroke. AB - BACKGROUND: Despite doubts about their efficacy and concern about their safety, antithrombotic agents are often used to treat acute ischemic stroke. Recent experience in patients with other thromboembolic disorders suggests that low molecular-weight heparin, which requires only subcutaneous administration once or twice daily, may be more effective and safer than standard (unfractionated) heparin. METHODS: We conducted a randomized, double-blind, placebo-controlled trial comparing two dosages of low-molecular-weight heparin with placebo in the treatment of ischemic stroke. Patients were randomly assigned within 48 hours of the onset of symptoms to receive high-dose nadroparin (4100 anti-factor Xa IU twice daily), low-dose nadroparin (4100 IU once daily), or placebo subcutaneously for 10 days. The primary measure of outcome was death or dependency regarding activities of daily living six months after randomization. Secondary outcomes were death, hemorrhagic transformation of the infarction, and other complications at 10 days, and death or dependency at 3 months. RESULTS: A total of 2750 patients were screened for the study. Among 312 patients randomized, 306 had outcomes that were analyzed at six months. Forty-five patients (45 percent) in the high-dose group, 53 patients (52 percent) in the low-dose group, and 68 patients (65 percent) in the placebo group died or became dependent. There was a significant dose-dependent effect among the three study groups in favor of low molecular-weight heparin (P = 0.005 by the chi-square test for trend). No significant differences among the groups in the occurrence of secondary outcomes were observed at 10 days. CONCLUSIONS: For patients with ischemic stroke treated within 48 hours of the onset of symptoms, low-molecular-weight heparin was effective in improving outcomes at six months. PMID- 7477194 TI - Cancer in the parents of children with cancer. AB - BACKGROUND: Certain types of cancer in children and young adults have been linked with an increased risk of cancer in close relatives. However, the relation between childhood cancer and familial risk remains to be fully assessed in population-based studies. METHODS: We conducted a nationwide study in Denmark of 11,380 parents of children with cancer. The children were identified from records in the Danish Cancer Registry; their parents were identified from population registers. The occurrence and rate of cancer in the parents were determined with use of the Cancer Registry's files and compared with national incidence rates for various categories of tumor. RESULTS: Overall, 1445 cancers were diagnosed in the parents, as compared with 1496 expected from national incidence rates, to yield standardized incidence ratios of 1.0 (95 percent confidence interval, 0.9 to 1.0) for all parents, 1.0 for mothers, and 0.9 for fathers. The lower rate of cancer among fathers reflected their lower standardized incidence ratio for lung cancer (0.8; 95 percent confidence interval, 0.6 to 0.9), as calculated from 144 observations. CONCLUSIONS: Genetic determinants are important in several types of childhood cancer, but the genetic susceptibility to tumors does not generally extend to the parents of children with cancer, nor do the patterns of incidence point to the influence of shared environmental factors. Thus, cancer in children should not be viewed as a general marker for an increased risk of cancer in the patients' parents. PMID- 7477196 TI - Images in clinical medicine. Right atrial tumor. PMID- 7477195 TI - Medication use and the risk of Stevens-Johnson syndrome or toxic epidermal necrolysis. AB - BACKGROUND: Toxic epidermal necrolysis and Stevens-Johnson syndrome are rare, life-threatening, drug-induced cutaneous reactions. We conducted a case-control study to quantify the risks associated with the use of specific drugs. METHODS: Data were obtained through surveillance networks in France, Germany, Italy, and Portugal. Drug use before the onset of disease was compared in 245 people who were hospitalized because of toxic epidermal necrolysis or Stevens-Johnson syndrome and 1147 patients hospitalized for other reasons (controls). Crude relative risks were calculated and adjusted for confounding by multivariate methods when numbers were large enough. RESULTS: Among drugs usually used for short periods, the risks were increased for trimethoprim-sulfamethoxazole and other sulfonamide antibiotics (crude relative risk, 172; 95 percent confidence interval, 75 to 396), chlormezanone (crude relative risk, 62; 21 to 188), aminopenicillins (multivariate relative risk, 6.7; 2.5 to 18), quinolones (multivariate relative risk, 10; 2.6 to 38), and cephalosporins (multivariate relative risk, 14; 3.2 to 59). For acetaminophen, the multivariate relative risk was 0.6 (95 percent confidence interval, 0.2 to 1.3) in France but 9.3 (3.9 to 22) in the other countries. Among drugs usually used for months or years, the increased risk was confined largely to the first two months of treatment, when crude relative risks were as follows: carbamazepine, 90 (95 percent confidence interval, 19 to infinity); phenobarbital, 45 (19 to 108); phenytoin, 53 (11 to infinity); valproic acid, 25 (4.3 to infinity); oxicam nonsteroidal antiinflammatory drugs (NSAIDs), 72 (25 to 209); allopurinol, 52 (16 to 167); and corticosteroids, 54 (23 to 124). For many drugs, including thiazide diuretics and oral hypoglycemic agents, there was no significant increase in risk. CONCLUSIONS: The use of antibacterial sulfonamides, anticonvulsant agents, oxicam NSAIDs, allopurinol, chlormezanone, and corticosteroids is associated with large increases in the risk of Stevens-Johnson syndrome or toxic epidermal necrolysis. But for none of the drugs does the excess risk exceed five cases per million users per week. PMID- 7477197 TI - Images in clinical medicine. Toxic epidermal necrolysis. PMID- 7477198 TI - Cardiac myxomas. AB - Although cardiac myxomas are histologically benign, they may be lethal because of their strategic position. They can mimic not only every cardiac disease but also infective, immunologic, and malignant processes. Myxomas must therefore be included in the differential diagnosis of valvular heart disease, cardiac insufficiency, cardiomegaly, bacterial endocarditis, disturbances of ventricular and supraventricular rhythm, syncope, and systemic or pulmonary embolism. The symptoms depend on the size, mobility, and location of the tumor. Echocardiography, including the transesophageal approach, is the most important means of diagnosis; CT and MRI may also be helpful. Coronary arteriography in patients over 40 years of age is generally required to rule out concomitant coronary artery disease. Surgical removal of the tumor should be performed as soon as possible; the long-term prognosis is excellent, and recurrences are rare. In follow-up examinations as well, echocardiography is essential. PMID- 7477199 TI - Community-acquired pneumonia. PMID- 7477200 TI - Case records of the Massachusetts General Hospital. Weekly clinicopathological exercises. Case 38-1995. A 68-year-old man with paresthesias and severe pain in both hands. PMID- 7477201 TI - Acute stroke--on the threshold of a therapy? PMID- 7477202 TI - Is there an inherited general susceptibility to cancer? PMID- 7477203 TI - To stay or not to stay? That is the question. PMID- 7477204 TI - Magnesium sulfate versus phenytoin for the prevention of eclampsia. PMID- 7477205 TI - Magnesium sulfate versus phenytoin for the prevention of eclampsia. PMID- 7477206 TI - Magnesium sulfate versus phenytoin for the prevention of eclampsia. PMID- 7477207 TI - Amiodarone in congestive heart failure. PMID- 7477208 TI - Amiodarone in congestive heart failure. PMID- 7477209 TI - Amiodarone in congestive heart failure. PMID- 7477210 TI - Conservative versus aggressive transfusion regimens in sickle cell disease. PMID- 7477211 TI - Conservative versus aggressive transfusion regimens in sickle cell disease. PMID- 7477212 TI - Endothelial nitric oxide synthase in the lungs of patients with pulmonary hypertension. PMID- 7477213 TI - A cystic fibrosis mutation associated with mild lung disease. PMID- 7477214 TI - Giant colonic diverticulum. PMID- 7477215 TI - More on the President's checkup. PMID- 7477216 TI - Women and children first. PMID- 7477217 TI - A preliminary trial of lamivudine for chronic hepatitis B infection. AB - BACKGROUND: Better treatments for chronic hepatitis B are needed. Lamivudine, the (-)enantiomer of 3'-thiacytidine, is a potent inhibitor of hepatitis B virus (HBV). METHODS: In a double-blind trial, we randomly assigned 32 patients with chronic hepatitis B (including 17 who had no response to earlier treatment with interferon) to receive 25, 100, or 300 mg of oral lamivudine daily for 12 weeks. The patients were then followed for 24 additional weeks. All the patients had hepatitis B antigen in serum. RESULTS: Levels of HBV DNA became undetectable (< or = 1.5 pg per milliliter) in 70 percent of the patients who received the 25-mg dose of lamivudine and 100 percent of those treated with the 100-mg or 300-mg dose. In most patients, HBV DNA reappeared after therapy was completed; however, six patients (19 percent), including five who had not responded to interferon, had sustained suppression of HBV DNA accompanied by normalization of alanine aminotransferase levels. Hepatitis B e antigen disappeared in four of these six patients (12 percent), three of whom had had no response to interferon. Levels of HBV DNA fell in all patients, including those who had had high levels at base line or normal alanine aminotransferase levels at base line, but sustained responses were more likely in patients with initially low HBV DNA levels and high alanine aminotransferase levels. During and after therapy, alanine aminotransferase levels at least doubled in five patients (50 percent) given the 25-mg dose and eight patients (36 percent) given the 100-mg or 300-mg dose. Minor adverse events occurred that were not related to the dose, as did transient, asymptomatic elevations of amylase, lipase, and creatine kinase levels. CONCLUSIONS: In a preliminary trial, 12 weeks of lamivudine therapy was well tolerated, and daily doses of 100 mg and 300 mg reduced HBV DNA to undetectable levels. PMID- 7477218 TI - Treatment with lamivudine, zidovudine, or both in HIV-positive patients with 200 to 500 CD4+ cells per cubic millimeter. North American HIV Working Party. AB - BACKGROUND: The reverse-transcriptase inhibitor lamivudine has in vitro synergy with zidovudine against the human immunodeficiency virus (HIV). We studied the activity and safety of lamivudine plus zidovudine as compared with either drug alone as treatment for patients with HIV infection, most of whom had not previously received zidovudine. METHODS: Three hundred sixty-six patients with 200 to 500 CD4+ cells per cubic millimeter who had received zidovudine for four weeks or less were randomly assigned to treatment with one of four regimens: 300 mg of lamivudine every 12 hours; 200 mg of zidovudine every 8 hours; 150 mg of lamivudine every 12 hours plus zidovudine; or 300 mg of lamivudine every 12 hours plus zidovudine. The study was double-blind and lasted 24 weeks, with an extension phase for another 28 weeks. RESULTS: Over the 24-week period, the low dose and high-dose regimens combining lamivudine and zidovudine were associated with greater increases in the CD4+ cell count (P = 0.002 and P = 0.015, respectively) and the percentage of CD4+ cells (P < 0.001 for both) and with greater decreases in plasma levels of HIV-1 RNA (P < 0.001 for both) than was treatment with zidovudine alone. Combination therapy was also more effective than lamivudine alone in lowering plasma HIV-1 RNA levels and increasing the percentage of CD4+ cells (P < 0.001 for all comparisons), and these advantages persisted through 52 weeks. Adverse events were no more frequent with combination therapy than with zidovudine alone. CONCLUSIONS: In HIV-infected patients with little or no prior antiretroviral therapy, treatment with a combination of lamivudine and zidovudine is well tolerated over a one-year period and produces more improvement in immunologic and virologic measures than does treatment with either agent alone. PMID- 7477219 TI - A clinical trial of the angiotensin-converting-enzyme inhibitor trandolapril in patients with left ventricular dysfunction after myocardial infarction. Trandolapril Cardiac Evaluation (TRACE) Study Group. AB - BACKGROUND: Treatment with angiotensin-converting-enzyme (ACE) inhibitors reduces mortality among survivors of acute myocardial infarction, but whether to use ACE inhibitors in all patients or only in selected patients is uncertain. METHODS: We screened 6676 consecutive patients with 7001 myocardial infarctions confirmed by enzyme studies. A total of 2606 patients had echocardiographic evidence of left ventricular systolic dysfunction (ejection fraction, < or = 35 percent). On days 3 to 7 after infarction, 1749 patients were randomly assigned to receive oral trandolapril (876 patients) or placebo (873 patients). The duration of follow-up was 24 to 50 months. RESULTS: During the study period, 304 patients (34.7 percent) in the trandolapril group died, as compared with 369 (42.3 percent) in the placebo group (P = 0.001). The relative risk of death in the trandolapril group, as compared with the placebo group, was 0.78 (95 percent confidence interval, 0.67 to 0.91). Trandolapril also reduced the risk of death from cardiovascular causes (relative risk, 0.75; 95 percent confidence interval, 0.63 to 0.89; P = 0.001) and sudden death (relative risk, 0.76; 95 percent confidence interval, 0.59 to 0.98; P = 0.03). Progression to severe heart failure was less frequent in the trandolapril group (relative risk, 0.71; 95 percent confidence interval, 0.56 to 0.89; P = 0.003). In contrast, the risk of recurrent myocardial infarction (fatal or nonfatal) was not significantly reduced (relative risk, 0.86; 95 percent confidence interval, 0.66 to 1.13; P = 0.29). CONCLUSIONS: Long term treatment with trandolapril in patients with reduced left ventricular function soon after myocardial infarction significantly reduced the risk of overall mortality, mortality from cardiovascular causes, sudden death, and the development of severe heart failure. That mortality was reduced in a randomized study enrolling 25 percent of consecutive patients screened should encourage the selective use of ACE inhibition after myocardial infarction. PMID- 7477220 TI - Images in clinical medicine. Paraganglioma of the middle ear. PMID- 7477221 TI - A national survey of the arrangements managed-care plans make with physicians. AB - BACKGROUND: Despite the growth of managed care in the United States, there is little information about the arrangements managed-care plans make with physicians. METHODS: In 1994 we surveyed by telephone 138 managed-care plans that were selected from 20 metropolitan areas nationwide. Of the 108 plans that responded, 29 were group-model or staff-model health maintenance organizations (HMOs), 50 were network or independent-practice-association (IPA) HMOs, and 29 were preferred-provider organizations (PPOs). RESULTS: Respondents from all three types of plan said they emphasized careful selection of physicians, although the group or staff HMOs tended to have more demanding requirements, such as board certification or eligibility. Sixty-one percent of the plans responded that physicians' previous patterns of costs or utilization of resources had little influence on their selection; 26 percent said these factors had a moderate influence; and 13 percent said they had a large influence. Some risk sharing with physicians was typical in the HMOs but rare in the PPOs. Fifty-six percent of the network or IPA HMOs used capitation as the predominant method of paying primary care physicians, as compared with 34 percent of the group or staff HMOs and 7 percent of the PPOs. More than half the HMOs reported adjusting payments according to utilization or cost patterns, patient complaints, and measures of the quality of care. Ninety-two percent of the network or IPA HMOs and 61 percent of the group or staff HMOs required their patients to select a primary care physician, who was responsible for most referrals to specialists. About three quarters of the HMOs and 31 percent of the PPOs reported using studies of the outcomes of medical care as part of their quality-improvement programs. CONCLUSIONS: Managed-care plans, particularly HMOs, have complex systems for selecting, paying, and monitoring their physicians. Hybrid forms are common, and the differences between group or staff HMOs and network or IPA HMOs are less extensive than is commonly assumed. PMID- 7477222 TI - The growth of medical groups paid through capitation in California. AB - BACKGROUND: In California, it is common for health maintenance organizations (HMOs) to contract with large medical groups that are paid through capitation and are responsible for managing a full spectrum of medical services. METHODS: We studied six large medical groups in California--Bristol Park Medical, Friendly Hills HealthCare Network, HealthCare Partners Medical Group, Mullikin Medical Centers, Palo Alto Medical Foundation, and San Jose Medical Group--that are paid through capitation and that are growing as a result of contracts with managed care organizations. We conducted interviews and obtained data on factors such as patient enrollment, capitation and other revenue, numbers of days spent by enrollees in the hospital, and numbers of visits to physicians per enrollee. RESULTS: Between 1990 and 1994, the number of HMO enrollees whose care was paid for through capitation in the six medical groups increased by 91 percent, from 398,359 to 759,474. In 1994, the mean number of hospital days per 1000 HMO enrollees ranged from 120 to 149 for non-Medicare patients and from 643 to 936 days for Medicare patients. By comparison, in 1993 the mean numbers of hospital days per 1000 HMO enrollees not covered by Medicare were 232 for California and 297 for the United States; for HMO enrollees covered by Medicare, the numbers were 1337 for California and 1698 for the United States. In 1994, the average annual number of visits to physicians for HMO patients in the six groups not covered by Medicare ranged from 3.1 to 3.9; for Medicare patients, it ranged from 7.2 to 9.3; these rates were slightly lower than statewide and national rates. Four of the groups have sold their assets (such as facilities, supplies, equipment, and patients' charts) to outside investors; the physicians remain employed by physician-owned professional corporations. CONCLUSIONS: Medical groups paid through capitation offer a model for the status of physicians in managed-care systems that differs from the employee status offered by staff-model HMOs and the subcontractor status offered by HMOs that negotiate directly with individual physicians. Despite their growth, such medical groups in California face substantial challenges, such as obtaining the financial assets necessary to sustain rapid growth. PMID- 7477223 TI - Drugs and thyroid function. PMID- 7477224 TI - Case records of the Massachusetts General Hospital. Weekly clinicopathological exercises. Case 39-1995. A 72-year-old man with exertional dyspnea, fatigue, and extensive ecchymoses and purpuric lesions. PMID- 7477225 TI - A quarter-century of antiviral therapy. PMID- 7477226 TI - Extreme risk--the new corporate proposition for physicians. PMID- 7477227 TI - Clinical problem-solving: diagnosing spousal abuse. PMID- 7477228 TI - Clinical problem-solving: diagnosing spousal abuse. PMID- 7477229 TI - Clinical problem-solving: diagnosing spousal abuse. PMID- 7477230 TI - Clinical problem-solving: diagnosing spousal abuse. PMID- 7477231 TI - Clinical problem-solving: diagnosing spousal abuse. PMID- 7477232 TI - Clinical problem-solving: diagnosing spousal abuse. PMID- 7477233 TI - Escherichia coli O157:H7. PMID- 7477234 TI - Escherichia coli O157:H7. PMID- 7477235 TI - Immunosuppressive therapy for myocarditis. PMID- 7477236 TI - Immunosuppressive therapy for myocarditis. PMID- 7477237 TI - Immunosuppressive therapy for myocarditis. PMID- 7477238 TI - Strut separations in Bjork-Shiley mitral valves. PMID- 7477239 TI - Soy protein and serum lipids. PMID- 7477240 TI - Laboratory-acquired Sabia virus infection. PMID- 7477242 TI - Effect of dietary intake of freeze dried bitter gourd (Momordica charantia) in streptozotocin induced diabetic rats. AB - Consumption of bitter gourd (Momordica charantia) by diabetic patients is a common practice in India, with the belief that it has an useful hypoglycemic potential. In the absence of conclusive information on the hypoglycemic influence of continuous intake of bitter gourd, in the present investigation, we have examined the hypoglycemic potency of dietary bitter gourd in experimentally induced diabetic rats. Wistar rats rendered hyperglycemic by streptozotocin (50 mg/kg b.w., i.p.) were maintained on a semi-synthetic diet containing freeze dried bitter gourd powder at 0.5% level for 6 weeks. The excretion of glucose, protein, urea and creatinine was monitored during the experimental period. Plasma glucose, albumin, urea and cholesterol were analysed at the end of the experimental regime. Dietary bitter gourd did not show any beneficial hypoglycemic influence as evidenced by the blood glucose levels as well as the excretion of diabetes related metabolites. PMID- 7477241 TI - Effect of intake of exogenous vitamins C, E and beta-carotene on the antioxidative status in kidneys of rats with streptozotocin-induced diabetes. AB - We studied the effect of supplementation with vitamins C, E and beta-carotene (PARABION, produced by Syndipharma) on antioxidative status in kidneys of male Wistar rats with diabetes induced by intravenous application of streptozotocin (45 mg.kg-1 of body weight). The animals received subtherapeutic doses of Insulin Interdep (6 U.kg-1 of body weight). A significant decrease of malondialdehyde (MDA), reduced (GSH) and oxidized (GSSG) glutathione and reduction of the activities of Se-glutathione peroxidase (Se-GSH-PX, EC. 1.11.1.9.) and glutathione S-transferase (GST, EC. 2.5.1.18.) were observed in kidneys of diabetic rats treated with these vitamins. On the contrary, the activity of CuZn superoxide dismutase (CuZn-SOD, EC. 1.15.1.1) and the level of vitamin C (vit. C) increased significantly. No changes were observed for vitamin E (vit. E), beta carotene and catalase (CAT, EC. 1.11.1.6). Supplementation with vitamins C, E and beta-carotene resulted in an improvement of antioxidative status of kidneys of rats with streptozotocin-induced diabetes. PMID- 7477243 TI - [The psychophysics of sweet taste. 7. New determination of sweet taste parameters of acesulfame, aspartame, cyclamate, saccharin, glucose and sorbitol]. AB - By means of a new method (double staircase) concentration pairs of equal intensity for the sweet compounds acesulfame-K, aspartame, cyclamate-Na, saccharine-Na, dextrose and glucitol were determined with sucrose on 6 or 7 equidistant scaled levels, respectively, with 4 repetitions. The parameters maximal intensity Rm, the concentration coefficient b and the mean Rm/b, respectively, are calculated for each substance from the known parameters for sucrose using the exponential function R = Rm(1-e-bS/Rm), which describes the sweetness intensity R in dependence of the concentration S. For exact determination of the maximal intensity the upper concentration level was fixed at very high degree. Additionally, the relative sweetness of each concentration level of each substance is calculated. In all cases the exponential function is valid without limitation. The statistical parameters are homogenous and are laying within the applied staircase method. Systematical deviations could not be found. PMID- 7477244 TI - Fermentation of rice-bengal gram dhal blends with whey: changes in phytic acid content and in vitro digestibility of starch and protein. AB - Whey fermentation of various rice and bengal gram dhal blends prepared by mixing them in different proportions at 35 degrees C for 18 h brought about a significant decline in phytic acid content. Phytic acid content in various blends decreased to the extent of 23 to 36 per cent over the control values. Whey incorporation as well as fermentation improved the starch and protein digestibility (in vitro) of all the rice-bengal gram dhal mixtures. Improvement in starch and protein digestibility is related to the reduction in phytic acid content, as this antinutrient is known to inhibit amylolysis and proteolysis. A significant negative correlation found between phytic acid and digestibility of starch and protein strengthens our findings. PMID- 7477245 TI - Aflatoxin-producing potential of Aspergillus flavus and Aspergillus parasiticus isolated from samples of smoked-dried meat. AB - Over a period of three years 420 samples of various smoke-dried meat products, collected from individual households in different region of Croatia were analysed for the presence of aflatoxigenic strains of the Aspergillus flavus group. Strains of A. flavus and A. parasiticus were present in 17.8% of the samples, and aflatoxin-producing ability was tested in 75 strains. In relation to sequential method of aflatoxin detection, 5 of 8 isolates were found in the first step (fluorescence in aflatoxin-producing ability medium--APA) and all of them in the second step (extraction method from syntheses on moist shredded wheat--SW). A. flavus strains produced mainly aflatoxin B1, and had various levels of toxigenicity (1.4-3.12 mg/kg). Some strains of A. parasiticus produced all four aflatoxins B1 B2 G1 G2, while the other ones produced AF B1 + G1 only, with concentrations of aflatoxins from 0.1 to 450 mg/kg. PMID- 7477247 TI - Whipping up panic about the pill. PMID- 7477246 TI - Chemical and sensory detection of diacetyl in strawberry juice (short communication). PMID- 7477248 TI - EMBO seeks stronger voice on policy. PMID- 7477249 TI - Health bodies urge backing for early warning procedures. PMID- 7477250 TI - SKB backs genome ethics programme. PMID- 7477251 TI - Assessing track records. PMID- 7477252 TI - Mind mechanisms. PMID- 7477253 TI - Born in summer? PMID- 7477254 TI - Duesberg and AIDS. PMID- 7477255 TI - Vision. Tuning retinal circuits. PMID- 7477256 TI - Human immunodeficiency virus. Trapped but still dangerous. PMID- 7477257 TI - Neurobiology. The bee's needs. PMID- 7477258 TI - Baboon fertility and social status. PMID- 7477259 TI - Vascular system defects and neuronal apoptosis in mice lacking ras GTPase activating protein. AB - The gene encoding p120-rasGAP, a negative regulator of Ras, has been disrupted in mice. This Gap mutation affects the ability of endothelial cells to organize into a highly vascularized network and results in extensive neuronal cell death. Mutati ons in the Gap and Nf1 genes have a synergistic effect, such that embryos homozygous for mutations in both genes show an exacerbated Gap phenotype. Thus rasGAP and neurofibromin act together to regulate Ras activity during embryonic development. PMID- 7477260 TI - Bee foraging in uncertain environments using predictive hebbian learning. AB - Recent work has identified a neuron with widespread projections to odour processing regions of the honeybee brain whose activity represents the reward value of gustatory stimuli. We have constructed a model of bee foraging in uncertain environments based on this type of neuron and a predictive form of hebbian synaptic plasticity. The model uses visual input from a simulated three dimensional world and accounts for a wide range of experiments on bee learning during foraging, including risk aversion. The predictive model shows how neuromodulatory influences can be used to bias actions and control synaptic plasticity in a way that goes beyond standard correlational mechanisms. Although several behavioural models of conditioning in bees have been proposed, this model is based on the neural substrate and was tested in a simulation of bee flight. PMID- 7477261 TI - Seeing motion behind occluders. AB - The visual system has no difficulty maintaining the identity of an object as it disappears and reappears behind stationary occluders. In the natural world, a moving object may differ from occluders by many characteristics (colour, depth, shape and so on). Scene segmentation based on these characteristics is thought to happen early in visual processing, and to influence how objects, including moving objects, are identified. What happens if the only characteristic distinguishing an object is its direction of motion? Experiments with random dot displays show that one dot moving in a constant trajectory is readily detected among identical dots in brownian motion. Detection declines sharply if the trajectory is intermittently broken, but improves if occluders obscure the breaks in the trajectory. It is not sufficient that these occluders be perceived as segmented from the rest of the display (such as by colour or depth). Rather, it is critical that the occluders do not contain motion that is similar in direction to that of the target trajectory. We conclude that detection of the trajectory is due to the integration of information within a network of low-level motion detectors and is not dependent on segmentation processes. PMID- 7477262 TI - Responses of cells in monkey visual cortex during perceptual filling-in of an artificial scotoma. AB - When we view a scene through one eye, we typically do not see the scotomas created by the optic disc and the blood vessels overlying the retinal surface. Similarly, when a texture field containing a hole is steadily viewed in peripheral vision (artificial scotoma), the hole appears to fill in with the surrounding texture in a matter of seconds, demonstrating that the visual system fills in information across regions where no information is available. Here we show that, in monkeys viewing a similar texture field with a hole, the responses of extrastriate visual neurons with receptive fields covering the hole increase gradually to a level comparable to that elicited by the same texture without a hole. The time course of these dynamic changes in activity parallels the time course of perceived filling-in of the hole by human observers, suggesting that this process mediates perceptual filling-in. PMID- 7477263 TI - Differential properties of two gap junctional pathways made by AII amacrine cells. AB - The retina is sensitive to light stimuli varying over more than 12 log units in intensity. It accomplishes this, in part, by switching between rod-dominated circuits designed for maximum utilization of scarce photons and cone circuits designed for greater acuity. Rod signals are integrated into the cone pathways through AII amacrine cells, which are connected by gap junctions both to other AII amacrine cells and to cone bipolar cells. To determine the relative permeabilities of the two junctional pathways, we have measured the distribution of biotinylated tracers across this heterologous cell assembly after injecting a single AII amacrine cell. We found that neurobiotin (relative molecular mass, 286) passed easily through both types of gap junctions, but that biotin-X cadaverine (relative molecular mass, 442) passed through AII/bipolar cell gap junctions poorly compared to AII/AII gap junctions. Thus, the AII/bipolar cell channel has a lower permeability to large molecules than does the AII/AII amacrine cell channel. The two pathways are also regulated differently. Dopamine and cyclic AMP agonists, known to diminish AII-AII coupling, did not change the relative labelling intensity of AII to bipolar cells. However, nitric oxide and cGMP agonists selectively reduced labelling in bipolar cells relative to AII amacrine cells, perhaps by acting at the bipolar side of this gap junction. This suggests that increased cGMP controls the network switching between rod and cone pathways associated with light adaptation. PMID- 7477264 TI - Unitary anion currents through phospholemman channel molecules. AB - Phospholemman (PLM) is a 72-amino-acid peptide with a single transmembrane domain, the expression of which induces chloride currents in Xenopus oocytes. It has remained unknown whether PLM is an ion channel or acts as a channel regulator. Here we show, by measuring unitary anion currents across planar phospholipid bilayers to which immunoaffinity-purified recombinant PLM was added, that it does indeed form ion channels. Excised patches of oocytes expressing PLM had similar currents. Of the ions tested, the sulphonic amino acid taurine was the most permeant, and expression of PLM increased fluxes of radiolabelled taurine in oocytes. Phospholemman is the smallest protein in cell membranes known to form an ion channel and the taurine selectivity suggests that it is involved in cell volume regulation. PMID- 7477265 TI - Follicular dendritic cells and human immunodeficiency virus infectivity. AB - Large amounts of human immunodeficiency virus (HIV) localize on follicular dendritic cells (FDC) in the follicles of secondary lymphoid tissues following viral infection. During clinical latency, active viral infection occurs primarily at these sites. As HIV on FDC is in the form of immune complexes, some of which may be formed with neutralizing antibody, we investigated whether HIV on FDC is infectious. We report here that HIV on FDC is highly infectious. Furthermore, FDC can convert neutralized HIV into an infectious form even in the presence of a vast excess of neutralizing antibody. Thus FDC may provide a mechanism whereby HIV infection can continue in the presence of neutralizing antibody. PMID- 7477266 TI - Behavioural and cardiovascular effects of disrupting the angiotensin II type-2 receptor in mice. AB - Angiotensin II, a potent regulator of blood pressure and of water and electrolyte balance, binds to two different G-protein-coupled receptors. The type-1 receptor (AT1) mediates the vasopressive and aldosterone-secreting effects of angiotensin II, but the function of the type-2 receptor (AT2) is unknown, although it is expressed in both adult and embryonic life. To address this question, we have generated mice lacking the gene encoding the AT2 receptor. Mutant mice develop normally, but have an impaired drinking response to water deprivation as well as a reduction in spontaneous movements. Their baseline blood pressure is normal, but they show an increased vasopressor response to injection of angiotensin II. Thus, although the AT2 receptor is not required for embryonic development, it plays a role in the central nervous system and cardiovascular functions that are mediated by the renin-angiotensin system. PMID- 7477267 TI - Effects on blood pressure and exploratory behaviour of mice lacking angiotensin II type-2 receptor. AB - There are two major angiotensin II receptor isoforms, AT1 and AT2. AT1 mediates the well-known pressor and mitogenic effects of angiotensin II, but the signalling mechanism and physiological role of AT2 has not been established. Its abundant expression in fetal tissues and certain brain nuclei suggest possible roles in growth, development and neuronal functions. Here we report the unexpected finding that the targeted disruption of the mouse AT2 gene resulted in a significant increase in blood pressure and increased sensitivity to the pressor action of angiotensin II. Thus AT2 mediates a depressor effect and antagonizes the AT1-mediated pressor action of angiotensin II. In addition, disruption of the AT2 gene attenuated exploratory behaviour and lowered body temperature. Our results show that angiotensin II activates AT1 and AT2, which have mutually counteracting haemodynamic effects, and that AT2 regulates central nervous system functions, including behaviour. PMID- 7477268 TI - Activation of the SAPK pathway by the human STE20 homologue germinal centre kinase. AB - Eukaryotic cells respond to different extracellular stimuli by recruiting homologous signalling pathways that use members of the MEKK, MEK and ERK families of protein kinases. The MEKK-->MEK-->ERK core pathways of Saccharomyces cerevisiae may themselves be regulated by members of the STE20 family of protein kinases. Here we report specific activation of the mammalian stress-activated protein kinase (SAPK) pathway by germinal centre kinase (GCK), a human STE20 homologue. SAPKs, members of the ERK family, are activated in situ by inflammatory stimuli, including tumour-necrosis factor (TNF) and interleukin-1, and phosphorylate and probably stimulate the transactivation function of c-Jun. Although GCK is found in many tissues, its expression in lymphoid follicles is restricted to the cells of the germinal centre, where it may participate in B cell differentiation. Activation of the SAPK pathway by GCK illustrates further the striking conservation of eukaryotic signalling mechanisms and defines the first physiological function of a mammalian Ste20. PMID- 7477269 TI - Initial hydrophobic collapse in the folding of barstar. AB - Two models are commonly used to describe the poorly understood earliest steps of protein folding. The framework model stresses very early formation of nascent secondary structures, which coalesce into a compact, molten, globule-like form from which tertiary structure slowly develops. The hydrophobic collapse model gives overriding precedence to a nonspecific collapse of the polypeptide chain which facilitates subsequent formation of specific secondary and tertiary structure. Here we report our analysis of the earliest observable events of the major folding pathway of barstar, a small protein. We compare the kinetics of folding using circular dichroism at 222 nm and 270 nm, intrinsic tryptophan fluorescence, fluorescence of the hydrophobic dye 8-anilino-1-naphthalene sulphonic acid on binding, and restoration of tryptophan-dansyl fluorescence energy transfer as structure-monitoring probes. We show that the polypeptide chain rapidly collapses (within 4 ms) to a compact globule with a solvent accessible hydrophobic core, but with no optically active secondary or tertiary structure. Thus the earliest event of the major folding pathway of barstar is a nonspecific hydrophobic collapse that does not involve concomitant secondary structure formation. PMID- 7477270 TI - Regulation of neural induction by the Chd and Bmp-4 antagonistic patterning signals in Xenopus. PMID- 7477271 TI - A new non-isotopic detection system for immunoassays. PMID- 7477272 TI - Contamination. PMID- 7477273 TI - Good manners. PMID- 7477274 TI - HIV and AIDS. PMID- 7477275 TI - Somatosensory cortex. Maps of time and space. PMID- 7477276 TI - Sexuality. Another important organ. PMID- 7477277 TI - Rooting the meristem. PMID- 7477278 TI - Genomics boosted as Japan unveils plans for state-funded companies. PMID- 7477279 TI - Geodesic surfactant structures. PMID- 7477280 TI - Evolution of deuterium on Venus. PMID- 7477281 TI - ESR ages for Krapina hominids. PMID- 7477282 TI - Structure of the fibre-forming protein pilin at 2.6 A resolution. AB - The crystallographic structure of Neisseria gonorrhoeae pilin, which assembles into the multifunctional pilus adhesion and virulence factor, reveals an alpha beta roll fold with a striking 85 A alpha-helical spine and an O-linked disaccharide. Key residues stabilize interactions that allow sequence hypervariability, responsible for pilin's celebrated antigenic variation, within disulphide region beta-strands and connections. Pilin surface shape, hydrophobicity and sequence variation constrain pilus assembly to the packing of flat subunit faces against alpha 1 helices. Helical fibre assembly is postulated to form a core of coiled alpha 1 helices banded by beta-sheet, leaving carbohydrate and hypervariable sequence regions exposed to solvent. PMID- 7477283 TI - Safety protocol due for a rough ride in biodiversity talks. PMID- 7477285 TI - Industry slow to invest in German biotech... PMID- 7477284 TI - Scaling of elastic strain energy in kangaroos and the benefits of being big. AB - Large kangaroos are unique among mammals in their ability to uncouple aerobic metabolic energy costs from the speed of locomotion, making hopping an economical gait. During the first half of the ground-contact phase, kinetic energy lost from the body is stored as elastic strain energy, predominantly in the hind limbs. The subsequent recoil returns kinetic and potential energy to the body. Here we show that the allometry of structures in the legs and feet of Macropodoidea is different from that of quadrupedal eutherian mammals. The potential for elastic energy storage in hoppers is shown to scale with strong positive allometry. This is a function of the structural properties of muscle-tendon units in the distal hind limbs and the postures adopted by hopping kangaroos. Our findings demonstrate how the use of tissue elasticity is strongly mass dependent and help explain the observed energetic phenomena. PMID- 7477286 TI - Optimism returns to biotech market as stocks recover. PMID- 7477287 TI - Cell fate in the Arabidopsis root meristem determined by directional signalling. AB - Postembryonic development in plants is achieved by apical meristems. Surgical studies and clonal analysis have revealed indirectly that cells in shoot meristems have no predictable destiny and that position is likely to play a role in the acquisition of cell identity. In contrast to animal systems, there has been no direct evidence for inductive signalling in plants until now. Here we present evidence for such signalling using laser ablation of cells in the root meristem of Arabidopsis thaliana. Although these cells show rigid clonal relationships, we now demonstrate that it is positional control that is most important in the determination of cell fate. Positional signals can be perpetuated from more mature to initial cells to guide the pattern of meristem cell differentiation. This offers an alternative to the general opinion that meristems are the source of patterning information. PMID- 7477289 TI - A sex difference in the human brain and its relation to transsexuality. AB - Transsexuals have the strong feeling, often from childhood onwards, of having been born the wrong sex. The possible psychogenic or biological aetiology of transsexuality has been the subject of debate for many years. Here we show that the volume of the central subdivision of the bed nucleus of the stria terminals (BSTc), a brain area that is essential for sexual behaviour, is larger in men than in women. A female-sized BSTc was found in male-to-female transsexuals. The size of the BSTc was not influenced by sex hormones in adulthood and was independent of sexual orientation. Our study is the first to show a female brain structure in genetically male transsexuals and supports the hypothesis that gender identity develops as a result of an interaction between the developing brain and sex hormones. PMID- 7477288 TI - Salt-resistant hypertension in mice lacking the guanylyl cyclase-A receptor for atrial natriuretic peptide. AB - Around half of all humans with essential hypertension are resistant to salt (blood pressure does not change by more than 5 mm Hg when salt intake is high), and although various inbred strains of rats display salt-insensitive elevated blood pressure, a gene defect to account for the phenotype has not been described. Atrial natriuretic peptide (ANP) is released from the heart in response to atrial stretch and is thought to mediate its natriuretic and vaso relaxant effects through the guanylyl cyclase-A receptor (GC-A). Here we report that disruption of the GC-A gene results in chronic elevations of blood pressure in mice on a normal salt diet. Unexpectedly, the blood pressure remains elevated and unchanged in response to either minimal or high salt diets. Aldosterone and ANP concentrations are not affected by the genotype. Therefore, mutations in the GC-A gene could explain some salt-resistant forms of essential hypertension and, coupled with previous work, further suggest that the GC-A signaling pathway dominates at the level of peripheral resistance, where it can operate independently of ANP. PMID- 7477290 TI - . . . but seeks "practical" gene therapy deals. PMID- 7477291 TI - Remodelling of hand representation in adult cortex determined by timing of tactile stimulation. AB - The primate somatosensory cortex, which processes tactile stimuli, contains a topographic representation of the signals it receives, but the way in which such maps are maintained is poorly understood. Previous studies of cortical plasticity indicated that changes in cortical representation during learning arise largely as a result of hebbian synaptic change mechanisms. Here we show, using owl monkeys trained to respond to specific stimulus sequence events, that serial application of stimuli to the fingers results in changes to the neuronal response specificity and maps of the hand surfaces in the true primary somatosensory cortical field (S1 area 3b). In this representational remodelling stimuli applied asychronously to the fingers resulted in these fingers being integrated in their representation, whereas fingers to which stimuli were applied asynchronously were segregated in their representation. Ventroposterior thalamus response maps derived in these monkeys were not equivalently reorganized. This representational plasticity appears to be cortical in origin. PMID- 7477292 TI - Synchronization of neuronal activity in hippocampus by individual GABAergic interneurons. AB - SYNCHRONIZATION of neuronal activity is fundamental in the operation of cortical networks. With respect to an ongoing synchronized oscillation, the precise timing of action potentials is an attractive candidate mechanism for information coding. Networks of inhibitory interneurons have been proposed to have a role in entraining cortical, synchronized 40-Hz activity. Here we demonstrate that individual GABAergic interneurons can effectively phase spontaneous firing and subthreshold oscillations in hippocampal pyramidal cells at 0 frequencies (4-7 Hz). The efficiency of this entrainment is due to interaction of GABAA-receptor mediated hyperpolarizing synaptic events with intrinsic oscillatory mechanisms tuned to this frequency range in pyramidal cells. Moreover, this GABAergic mechanism is sufficient to synchronize the firing of pyramidal cells. Thus, owing to the divergence of each GABAergic interneuron, more than a thousand pyramidal cells may share a common temporal reference established by an individual interneuron. PMID- 7477293 TI - Mechanosensory signalling in C. elegans mediated by the GLR-1 glutamate receptor. AB - NEURONAL signalling across synapses involves activation of many neurotransmitter receptors on postsynaptic cells. glr-1 encodes a potential glutamate receptor in the nematode Caenorhabditis elegans which is most similar to vertebrae AMPA-type ionotropic glutamate receptors. glr-1 is expressed in motor neurons and interneurons, including interneurons implicated in the control of locomotion. Here we investigate the contribution of glr-1 to the normal signalling of these neurons, by generating a deletion mutation in glr-1. We find that mutant worms are deficient in their ability to withdraw backwards when mechanically stimulated, but they withdraw normally in response to chemical repellents. The ASH sensory neurons mediate withdrawal responses both to mechanical stimuli and to repellents, and ASH makes chemical synapses with glr-1-expressing interneurons. Our results suggest that postsynaptic interneurons use different neurotransmitter receptors to process two sensory stimuli detected by one sensory neuron. PMID- 7477294 TI - Synaptic code for sensory modalities revealed by C. elegans GLR-1 glutamate receptor. AB - How does the nervous system encode environmental stimuli as sensory experiences? Both the type (visual, olfactory, gustatory, mechanical or auditory) and the quality of a stimulus (spatial position, intensity or frequency) are represented as a neural code. Here we undertake a genetic analysis of sensory modality coding in Caenorhabditis elegans. The ASH sensory neurons respond to two distinct sensory stimuli (nose touch and osmotic stimuli). A mutation in the glr-1 (glutamate receptor) gene eliminates the response to nose touch but not to osmotic repellents. The predicted GLR-1 protein is roughly 40% identical to mammalian AMPA-class glutamate receptor (GluR) subunits. Analysis of glr-1 expression and genetic mosaics indicates that GLR-1 receptors act in synaptic targets of the ASH neurons. We propose that discrimination between the ASH sensory modalities arises from differential release of ASH neurotransmitters in response to different stimuli. PMID- 7477297 TI - Accidental contamination 'unlikely'. PMID- 7477295 TI - Clustering of Shaker-type K+ channels by interaction with a family of membrane associated guanylate kinases. AB - ANCHORING of ion channels at specific subcellular sites is critical for neuronal signalling, but the mechanisms underlying channel localization and clustering are largely unknown (reviewed in ref. 1). Voltage-gated K+ channels are concentrated in various neuronal domains, including presynaptic terminals, nodes of Ranvier and dendrites, where they regulate local membrane excitability. Here we present functional and biochemical evidence that cell-surface clustering of Shaker subfamily K+ channels is mediated by the PSD-95 family of membrane-associated putative guanylate kinases, as a result of direct binding of the carboxy-terminal cytoplasmic tails to the K+ channel subunits to two PDZ (also known as GLGF or DHR) domains in the PSD-95 protein. The ability of PDZ domains to function as independent modules for protein-protein interaction, and their presence in other junction-associated molecules (such as ZO-1 (ref. 3) and syntrophin), suggest that PDZ-domain-containing polypeptides may be widely involved in the organization of proteins at sites of membrane specialization. PMID- 7477298 TI - HIV-blood scientist defends reporting delay. PMID- 7477296 TI - Cloning of a new cytokine that induces IFN-gamma production by T cells. AB - The mechanism underlying the differentiation of CD4+ T cells into functionally distinct subsets (Th1 and Th2) is incompletely understood, and hitherto unidentified cytokines may be required for the functional maturation of these cells. Here we report the cloning of a recently identified IFN-gamma-inducing factor (IGIF) that augments natural killer (NK) activity in spleen cells. The gene encodes a precursor protein of 192 amino acids and a mature protein of 157 amino acids, which have no obvious similarities to any peptide in the databases. Messenger RNAs for IGIF and interleukin-12 (IL-12) are readily detected in Kupffer cells and activated macrophages. Recombinant IGIF induces IFN-gamma more potently than does IL-12, apparently through a separate pathway. Administration of anti-IGIF antibodies prevents liver damage in mice inoculated with Propionibacterium acnes and challenged with lipopolysaccharide, which induces toxic shock. IGIF may be involved in the development of Th1 cells and also in mechanisms of tissue injury in inflammatory reactions. PMID- 7477299 TI - A eukaryotic transcriptional repressor with carboxypeptidase activity. AB - Adipocyte differentiation involves the transcriptional activation of several genes in triglyceride metabolism, including the adipose P2 (aP2 or 422) gene that encodes the adipocyte lipid-binding protein ALBP. Within the mouse aP2 promoter region, the AE-1 sequence functions as either a positive or a negative element in the regulation of aP2 gene expression. The AE-1 sequence is the binding site for the positive murine (3T3) adipocyte factor C/EBP-alpha, several human preadipocyte factors, and a 3T3 preadipocyte factor(s) that has been implicated as a repressor of aP2 gene expression. Here we report the cloning of new complementary DNAs that encode the 3T3 preadipocyte factor (termed AEBP1) and demonstrate that AEBP1 expression is abolished during adipocyte differentiation. Furthermore, we show that an activity of a carboxypeptidase associated with AEBP1 is important in the transcriptional repression function of AEBP1. Thus AEBP1 might represent a new type of transcription factor that regulates transcription by cleavage of factors involved in transcription. PMID- 7477301 TI - Advisors may urge NIH to seek out industry support for clinical trials. PMID- 7477300 TI - N-glycans as apical sorting signals in epithelial cells. AB - In epithelial Madin-Darby canine kidney (MDCK) cells newly synthesized molecules are sorted in the trans-Golgi network and directly delivered to their apical and basolateral surface destinations. Sorting is mediated by signals in the cytoplasmic domains of basolateral transmembrane proteins whereas glycosylphosphatidylinositol-linked proteins have apical sorting information in their glycolipid tails. Signals for apical transmembrane proteins are thought to reside in their ectodomains, because truncated forms lacking the cytoplasmic tail and the membrane anchor are secreted apically. Here we demonstrate that carbohydrates act as an apical targeting signal for secretory proteins. Growth hormone, which is non-glycosylated and secreted from both sides of MDCK cell layers, is secreted from the apical side when glycosylated. Thus glycans not only play a general role in protein folding but also appear to function in protein sorting in biosynthetic traffic. PMID- 7477302 TI - Managed health-care adds to woes of US medical schools. PMID- 7477303 TI - Wellcome sets sequencing project in motion. PMID- 7477304 TI - Genetic testing 'needs more checks'. PMID- 7477305 TI - Eugenics row inflames vote on faculty name. PMID- 7477306 TI - Causes of death in haemophilia. PMID- 7477307 TI - Nicotine as a drug. PMID- 7477308 TI - Tonegawa defended. PMID- 7477309 TI - Benefits of placebos. PMID- 7477310 TI - Benefits of placebos. PMID- 7477311 TI - How trinucleotide repeats may function. PMID- 7477312 TI - Schizophrenia. Charting the circuits. PMID- 7477313 TI - Spatial learning. The LTP-memory connection. PMID- 7477314 TI - Macromolecular physiology. One small step for myosin... PMID- 7477315 TI - Haem polymerization in malaria. PMID- 7477316 TI - Gene duplications in H. influenzae. PMID- 7477318 TI - A functional neuroanatomy of hallucinations in schizophrenia. AB - Hallucinations, perceptions in the absence of external stimuli, are prominent among the core symptoms of schizophrenia. The neural correlates of these brief, involuntary experiences are not well understood, and have not been imaged selectively. We have used new positron emission tomography (PET) methods to study the brain state associated with the occurrence of hallucinations in six schizophrenic patients. Here we present a group study of five patients with classic auditory verbal hallucinations despite medication, demonstrating activations in subcortical nuclei (thalamic, striatal), limbic structures (especially hippocampus), and paralimbic regions (parahippocampal and cingulate gyri, as well as orbitofrontal cortex). We also present a case study of a unique, drug-naive patient with visual as well as auditory verbal hallucinations, demonstrating activations in visual and auditory/linguistic association cortices as part of a distributed cortical-subcortical network. Activity in deep brain structures, identified with group analysis, may generate or modulate hallucinations, and the particular neocortical regions entrained in individual patients may affect their specific perceptual content. The interaction of these distributed neural systems provides a biological basis for the bizarre reports of schizophrenic patients. PMID- 7477317 TI - A homeobox gene essential for zebrafish notochord development. AB - The notochord is a midline mesodermal structure with an essential patterning function in all vertebrate embryos. Zebrafish floating head (flh) mutants lack a notochord, but develop with prechordal plate and other mesodermal derivatives, indicating that flh functions specifically in notochord development. We show that floating head is the zebrafish homologue of Xnot, a homeobox gene expressed in the amphibian organizer and notochord. We propose that flh regulates notochord precursor cell fate. PMID- 7477319 TI - Dopaminergic modulation of impaired cognitive activation in the anterior cingulate cortex in schizophrenia. AB - Dopaminergic dysregulation remains an empirical cornerstone for theories concerning the causation of schizophrenia. Evidence for a dopamine system dysfunction in schizophrenia includes the psychosis-inducing effects of dopaminergic agonists and the antipsychotic potency of dopaminergic antagonists. Here we use positron emission tomography (PET) to examine the regulatory role of dopamine on cortical function in normal subjects and unmedicated schizophrenic patients. Using a factorial experimental design, we compared the effect of dopaminergic manipulation with apomorphine on a neural response to a cognitive task. In the schizophrenic patients, relative to controls, an impaired cognitive activation of the anterior cingulate cortex was significantly modulated by a manipulation of dopaminergic neurotransmission. Thus, after apomorphine, the schizophrenic subjects displayed a significantly enhanced cognitive activation of the anterior cingulate cortex relative to the controls. These data provide in vivo evidence that an impaired cognitive-task-induced activation of the anterior cingulate cortex in schizophrenic patients can be significantly modulated by a dopaminergic manipulation. PMID- 7477320 TI - Distinct components of spatial learning revealed by prior training and NMDA receptor blockade. AB - Synaptic plasticity dependent on N-methyl-D-aspartate (NMDA) receptors is thought to underlie certain types of learning and memory. In support of this, both hippocampal long-term potentiation and spatial learning in a watermaze are impaired by blocking NMDA receptors with a selective antagonist D(-)-2-amino-5 phosphonovaleric acid (AP5) or by a mutation in one of the receptor subunits. Here we report, however, that the AP5-induced learning deficit can be almost completely prevented if rats are pretrained in a different watermaze before administration of the drug. This is not because of stimulus generalization, and occurs despite learning of the second task remaining hippocampus dependent. An AP5-induced learning deficit is, however, still seen if the animals are pretrained using a non-spatial task. Thus, despite its procedural simplicity, the watermaze may involve multiple cognitive processes with distinct pharmacological properties; although required for some component of spatial learning, NMDA receptors may not be required for encoding the spatial representation of a specific environment. PMID- 7477321 TI - Spatial learning without NMDA receptor-dependent long-term potentiation. AB - Hippocampal lesions impair spatial learning in the watermaze. Drugs that antagonize N-methyl-D-aspartate (NMDA)-receptor activity, which is required for long-term potentiation (LTP) at various hippocampal synapses, block LTP and impair watermaze learning. This has led to the hypothesis that NMDA receptors, through their involvement in LTP, may be necessary for spatial and other forms of learning. We examined this hypothesis using NPC17742 (2R,4R,5S-2-amino-4,5-(1,2 cyclo hexyl)-7-phosphonoheptano acid), a potent and specific antagonist of NMDA receptors. Here we report that NPC17742 completely blocked dentate gyrus LTP but did not prevent normal spatial learning in rats that had been made familiar with the general task requirements by non-spatial pretraining. Although these results do not rule out a contribution of NMDA-mediated dentate LTP to spatial learning, they indicate that this form of LTP is not required for normal spatial learning in the watermaze. PMID- 7477322 TI - NT-4-mediated rescue of lateral geniculate neurons from effects of monocular deprivation. AB - Altering the balance of activity between the two eyes during the critical period for visual-system development profoundly affects competitive interactions among neurons in the lateral geniculate nucleus and primary visual cortex. Neurons in the lateral geniculate nucleus that are deprived of activity by closing or silencing one eye atrophy as a result of competition with non-deprived neurons for some critical factor(s) presumed to be present in the cortex. Based on their actions in the developing visual system, neurotrophins are attractive candidates for such factors. We tested whether neurotrophins mediate intracortical competition of afferents from the lateral geniculate nucleus by using monocular deprivation and a new method for highly localized, in vivo delivery of neurotrophins. This method allowed unambiguous identification of neurons that were exposed to neurotrophin. Here we report that only one neurotrophin, the TrkB ligand NT-4, rescued neurons in the lateral geniculate nucleus from the dystrophic effects of monocular deprivation. PMID- 7477323 TI - Effects of brain-derived neurotrophic factor on optic axon branching and remodelling in vivo. AB - Neurotrophins are thought to be important for the survival and differentiation of vertebrate neurons. Roles have been suggested for target-derived neurotrophins, based both on their expression in target tissues at the time of neuron innervation, and on their effects on axonal sprouting. However, direct in vivo evidence of their involvement in axon arborization has remained elusive. We have used in vivo microscopy to follow individual optic axons over time, and have examined the role of the neurotrophin brain-derived neurotrophic factor (BDNF) in their development. Here we show that injection of BDNF into the optic tectum of live Xenopus laevis tadpoles increased the branching and complexity of optic axon terminal arbors. In contrast, injection of specific neutralizing antibodies to BDNF reduced axon arborization and complexity. The onset of these effects was rapid (within 2 hours) and persisted throughout the 24-hour observation period. Other neurotrophins had little or no significant effects. These results demonstrate the involvement of neurotrophins in the dynamic elaboration of axon terminals, and suggest a direct role for target-derived BDNF during synaptic patterning in the developing central nervous system. PMID- 7477324 TI - Defective recycling of synaptic vesicles in synaptotagmin mutants of Caenorhabditis elegans. AB - Synaptotagmin, an integral membrane protein of the synaptic vesicle, binds calcium and interacts with proteins of the plasma membrane. These observations suggest several possible functions for synaptotagmin in synaptic vesicle dynamics: it could facilitate exocytosis by promoting calcium-dependent fusion, inhibit exocytosis by preventing fusion, or facilitate endocytosis of synaptic vesicles from the plasma membrane by acting as a receptor for the endocytotic proteins of the clathrin AP2 complex. Here we show that synaptic vesicles are depleted at synaptic terminals in synaptotagmin mutants of the nematode Caenorhabditis elegans. This depletion is not caused by a defect in transport or by increased synaptic vesicle release, but rather by a defect in retrieval or synaptic vesicles from the plasma membrane. Thus we propose that, as well as being involved in exocytosis, synaptotagmin functions in vesicular recycling. PMID- 7477325 TI - Role of SUPERMAN in maintaining Arabidopsis floral whorl boundaries. AB - The Arabidopsis gene SUPERMAN (SUP) is necessary for the proper spatial development of reproductive floral tissues. Recessive mutations cause extra stamens to form interior to the normal third whorl stamens, at the expense of fourth whorl carpel development. The mutant phenotype is associated with the ectopic expression of the B function genes, AP3 and PI, in the altered floral region, closer to the centre of the flower than in the wild type, and ap3 sup and pi sup double mutants exhibit a phenotype similar to ap3 and pi single mutants. These findings led to SUP being interpreted as an upstream negative regulator of the B function organ-identity genes, acting in the fourth whorl, to establish a boundary between stamen and carpel whorls. Here we show, using molecular cloning and analysis, that it is expressed in the third whorl and acts to maintain this boundary in developing flowers. The putative SUPERMAN protein contains one zinc finger and a region resembling a basic leucine zipper motif, suggesting a function in transcriptional regulation. PMID- 7477326 TI - Rescue of early embryonic lethality in mdm2-deficient mice by deletion of p53. AB - The gene p53 encodes a transcriptional activator of genes involved in growth arrest, DNA repair and apoptosis. Loss of p53 function contributes to tumour development in vivo. The transcriptional activation function of p53 is inactivated by interaction with the mdm2 gene product. Amplification of mdm2 has been observed in 36% of human sarcomas, indicating that it may represent an alternative mechanism of preventing p53 function in tumour development. To study mdm2 function in vivo, we generated an mdm2 null allele by homologous recombination. Mdm2 null mice are not viable, and further analysis revealed embryonic lethality around implantation. To examine the importance of the interaction of MDM2 with p53 in vivo, we crossed mice heterozygous for mdm2 and p53 and obtained progeny homozygous for both p53 and mdm2 null alleles. Rescue of the mdm2-/- lethality in a p53 null background suggests that a critical in vivo function of MDM2 is the negative regulation of p53 activity. PMID- 7477327 TI - Rescue of embryonic lethality in Mdm2-deficient mice by absence of p53. AB - The Mdm2 proto-oncogene was originally identified as one of several genes contained on a mouse double minute chromosome present in a transformed derivative of 3T3 cells. Overexpression of Mdm2 can immortalize primary cultures of rodent fibroblasts. Human MDM2 is amplified in 30-40% of sarcomas, and is overexpressed in leukaemic cells. The Mdm2 oncoprotein forms a complex with the p53 tumour suppressor protein and inhibits p53-mediated transregulation of gene expression. Because Mdm2 expression increases in response to p53, Mdm2-p53 binding may autoregulate Mdm2 expression and modulate the activity of p53 in the cell. We have created Mdm2-null and Mdm2/p53-null mice to determine whether Mdm2 possesses developmental functions in addition to the ability to complex with p53, and to investigate the biological role of Mdm2-p53 complex formation in development. Mice deficient for Mdm2 die early in development. In contrast, mice deficient for both Mdm2 and p53 develop normally and are viable. These results suggest that a critical role of Mdm2 in development is the regulation of p53 function. PMID- 7477328 TI - Movement and force produced by a single myosin head. AB - Muscle contraction is driven by the cyclical interaction of myosin with actin, coupled to the breakdown of ATP. Studies of the interaction of filamentous myosin and of a double-headed proteolytic fragment, heavy meromyosin (HMM), with actin have demonstrated discrete mechanical events, arising from stochastic interaction of single myosin molecules with actin. Here we show, using an optical-tweezers transducer, that a single myosin subfragment-1 (S1), which is a single myosin head, can act as an independent generator of force and movement. Our analysis accounts for the broad distribution of displacement amplitudes observed, and indicates that the underlying movement (working stroke) produced by a single acto S1 interaction is approximately 4 nm, considerably shorter than previous estimates but consistent with structural data. We measure the average force generated by S1 or HMM to be at least 1.7 pN under isometric conditions. PMID- 7477329 TI - A potential catalytic site revealed by the 1.7-A crystal structure of the amino terminal signalling domain of Sonic hedgehog. AB - Within the past few years, members of the hedgehog (hh) family of secreted signalling proteins have emerged as the primary signals generated by certain embryonic patterning centres. In vertebrate embryos, for example, sonic hedgehog expression in the notochord appears to be responsible for the local and long range induction of ventral cell types within the neural tube and somites (reviewed in refs 1, 2). Protein products encoded by hh family members are synthesized as precursors that undergo autoprocessing to generate an amino terminal domain that appears to be responsible for both local and long-range signalling activities, and a carboxy-terminal domain that contains the autoprocessing activity. As part of an effort to understand how hh family members participate in cell-to-cell signalling, we have determined and report here the crystal structure at 1.7 A of the amino-terminal domain of murine Sonic hedgehog (Shh-N). The structure revealed a tetrahedrally coordinated zinc ion that appears to be structurally analogous to the zinc coordination sites of zinc hydrolases, such as thermolysin and carboxypeptidase A. This previously unsuspected catalytic site represents a distinct activity from the autoprocessing activity that resides in the carboxy-terminal domain. PMID- 7477330 TI - Howard Hughes to shift focus of support away from institutions. PMID- 7477331 TI - France backs off promise to boost UN AIDS programme. PMID- 7477332 TI - Geneticist quits in protest at 'genes and violence' claim. PMID- 7477333 TI - 'Genetic art' builds cryptic bridge between two cultures. PMID- 7477334 TI - Historic apoptosis. PMID- 7477335 TI - Corticotropin-releasing factor. From stress to cognition. PMID- 7477336 TI - Genome sequences. A microbial minimalist. PMID- 7477337 TI - DNA models. Mutations caught in the act. PMID- 7477338 TI - Australopithecus goes west. PMID- 7477339 TI - Palaeoanthropology. Out of Africa and into Asia. PMID- 7477340 TI - Protecting HIV databases. PMID- 7477341 TI - The enigma of the natural killer cell. AB - Natural killer (NK) cells are controlled by receptors specific for polymorphic determinants of class I molecules of the major histocompatibility complex (MHC). The contrasting properties of NK and cytotoxic T cell (CTL) class I receptors provide complementarity in the cytolytic lymphocyte response to viruses, tumours and transplants. Whereas human NK cell class I receptors consist of immunoglobulin domains, their mouse counterparts resemble C-type lectins. This difference may reflect the receptors' diverse and rapidly evolving class I ligands. PMID- 7477342 TI - Femtosecond molecular dynamics of tautomerization in model base pairs. AB - Hydrogen bonds commonly lend robustness and directionality to molecular recognition processes and supramolecular structures. In particular, the two or three hydrogen bonds in Watson-Crick base pairs bind the double-stranded DNA helix and determine the complementarity of the pairing. Watson and Crick pointed out, however, that the possible tautomers of base pairs, in which hydrogen atoms become attached to the donor atom of the hydrogen bond, might disturb the genetic code, as the tautomer is capable of pairing with different partners. But the dynamics of hydrogen bonds in general, and of this tautomerization process in particular, are not well understood. Here we report observations of the femtosecond dynamics of tautomerization in model base pairs (7-azaindole dimers) containing two hydrogen bonds. Because of the femtosecond resolution of proton motions, we are able to examine the cooperativity of formation of the tautomer (in which the protons on each base are shifted sequentially to the other base), and to determine the characteristic timescales of the motions in a solvent-free environment. We find that the first step occurs on a timescale of a few hundred femtoseconds, whereas the second step, to form the full tautomer, is much slower, taking place within several picoseconds; the timescales are changed significantly by replacing hydrogen with deuterium. These results establish the molecular basis of the dynamics and the role of quantum tunnelling. PMID- 7477343 TI - Cell-free synthesis of polyketides by recombinant erythromycin polyketide synthases. AB - Modular polyketide synthases (PKSs) are complex multi-enzyme proteins that catalyse the bacterial biosynthesis of many pharmaceutically useful polyketides. The PKSs are organized into a series of modules, each containing the active catalytic sites required for one step in the synthesis process. Here we report a method for cell-free enzymatic synthesis of 6-deoxyerythronolide B (6-dEB), the parent molecule of the antibiotic erythromycin A, using recombinant 6 deoxyerythronolide B synthase (DEBS), a modular PKS with at least 28 distinct active sites. We have also synthesized in vitro a triketide lactone by using a truncated mutant of DEBS. The availability of such cell-free synthetic routes will allow direct investigation of the structural and mechanistic basis for the unusual combination of high substrate specificity and tolerance to genetic reprogramming found in this enzyme family. PMID- 7477344 TI - The first australopithecine 2,500 kilometres west of the Rift Valley (Chad) AB - The first sites with Pliocene and Pleistocene mammals west of the Rift Valley in Central Africa in northern Chad were reported in 1959 (ref. 1), and documented the presence of mixed savannah and woodland habitats. Further sites and a probable Homo erectus cranio-facial fragment were subsequently discovered. In 1993 a survey of Pliocene and Pleistocene formations in the Borkou-Ennedi-Tibesti Province of Chad (B.E.T.) led to the discovery of 17 new sites in the region of Bahr el Ghazal (classical Arabic for River of the Gazelles) near Koro Toro. One site, KT 12 (15 degrees 58'10"N, 18 degrees 52'46"E) yielded an australopithecine mandible associated with a fauna biochronologically estimated to be 3.0-3.5 Myr old. Australopithecine species described since 1925 are known from southern Africa and from sites spread along the eastern Rift Valley from Tanzania to Ethiopia (Fig. 1). This new find from Chad, which is most similar in morphology to Australopithecus afarensis, documents the presence of an early hominid a considerable distance, 2,500 km, west of the Rift Valley. PMID- 7477345 TI - Early Homo and associated artefacts from Asia. AB - The site of Longgupo Cave was discovered in 1984 and excavated in 1985-1988 by the Institute of Vertebrate Paleontology and Paleoanthropology (Beijing) and the Chongqing National Museum (Sichuan Province). Important finds include very archaic hominid dental fragments, Gigantopithecus teeth and primitive stone tools. Palaeomagnetic analysis and the presence of Ailuropoda microta (pygmy giant panda) suggested that the hominid-bearing levels dated to the earliest Pleistocene. In 1992, joint Chinese-American-Canadian geochronological research corroborated the age using electron spin resonance (ESR) analysis. We report here that the hominid dentition and stone tools from Longgupo Cave are comparable in age and morphology with early representives of the genus Homo (H. habilis and H. ergaster) and the Oldowan technology in East Africa. The Longgupo dentition is demonstrably more primitive than that seen in Asian Homo erectus. Longgupo's diverse and well preserved Plio-Pleistocene fauna of 116 species provide a sensitive contextual base for interpreting the early arrival of the genus Homo in Asia. PMID- 7477346 TI - The neural basis of the central executive system of working memory. AB - Working memory refers to a system for temporary storage and manipulation of information in the brain, a function critical for a wide range of cognitive operations. It has been proposed that working memory includes a central executive system (CES) to control attention and information flow to and from verbal and spatial short-term memory buffers. Although the prefrontal cortex is activated during both verbal and spatial passive working memory tasks, the brain regions involved in the CES component of working memory have not been identified. We have used functional magnetic resonance imaging (fMRI) to examine brain activation during the concurrent performance of two tasks, which is expected to engage the CES. Activation of the prefrontal cortex was observed when both tasks are performed together, but not when they are performed separately. These results support the view that the prefrontal cortex is involved in human working memory. PMID- 7477347 TI - Specificity of monosynaptic connections from thalamus to visual cortex. AB - In cortical area 17 of the cat, simple receptive fields are arranged in elongated subregions that respond best to bright (on) or dark (off) oriented contours, whereas the receptive fields of their thalamic inputs have a concentric on and off organization. This dramatic transformation suggests that there are specific rules governing the connections made between thalamic and cortical neurons (see ref. 4). Here we report a study of these rules in which we recorded from thalamic (lateral geniculate nucleus; LGN) and cortical neurons simultaneously and related their receptive fields to their connectivity, as measured by cross-correlation analysis. The probability of finding a monosynaptic connection was high when a geniculate receptive field was superimposed anywhere over an elongated simple cell subregion of the same signature (on or off). However, 'inappropriate' connections from geniculate cells of the opposite receptive field signature were extremely rare. Together, these findings imply that the outline of the elongated, simple receptive field, and thus of cortical orientation selectivity, is laid down at the level of the first synapse from the thalamic afferents. PMID- 7477348 TI - Displacement of corticotropin releasing factor from its binding protein as a possible treatment for Alzheimer's disease. AB - In Alzheimer's disease (AD) there are dramatic reductions in the content of corticotropin releasing factor (CRF), reciprocal increases in CRF receptors, and morphological abnormalities in CRF neurons in affected brain areas. Cognitive impairment in AD patients is associated with a lower cerebrospinal fluid concentration of CRF, which is known to induce increases in learning and memory in rodents. This suggests that CRF deficits contribute to cognitive impairment. The identification in post-mortem brain of CRF-binding protein (CRF-BP), a high affinity binding protein that inactivates CRF, and the differential distribution of CRF-BP and CRF receptors, provides the potential for improving learning and memory without stress effects of CRF receptor agonists. Here we show that ligands that dissociate CRF from CRF-BP increase brain levels of 'free CRF' in AD to control levels and show cognition-enhancing properties in models of learning and memory in animals without the characteristic stress effects of CRF receptor agonists. PMID- 7477349 TI - Urocortin, a mammalian neuropeptide related to fish urotensin I and to corticotropin-releasing factor. AB - Corticotropin-releasing factor (CRF), a peptide first isolated from mammalian brain, is critical in the regulation of the pituitary-adrenal axis, and in complementary stress-related endocrine, autonomic and behavioural responses. Fish urotensin I and amphibian sauvagine were considered to be homologues of CRF until peptides even more closely related to CRF were identified in these same vertebrate classes. We have characterized another mammalian member of the CRF family and have localized its urotensin-like immunoreactivity to, and cloned related complementary DNAs from, a discrete rat midbrain region. The deduced protein encodes a peptide that we name urocortin, which is related to urotensin (63% sequence identity) and CRF (45% sequence identity). Synthetic urocortin evokes secretion of adrenocorticotropic hormone (ACTH) both in vitro and in vivo and binds and activates transfected type-1 CRF receptors, the subtype expressed by pituitary corticotropes. The coincidence of urotensin-like immunoreactivity with type-2 CRF receptors in brain, and our observation that urocortin is more potent than CRF at binding and activating type-2 CRF receptors, as well as at inducing c-Fos (an index of cellular activation) in regions enriched in type-2 CRF receptors, indicate that this new peptide could be an endogenous ligand for type-2 CRF receptors. PMID- 7477350 TI - A stomatin-like protein necessary for mechanosensation in C. elegans. AB - The mec-2 gene is required for the function of a set of six touch receptor neurons in the nematode Caenorhabditis elegans; mec-2 mutants, which are touch insensitive, have touch cells that appear morphologically normal. Gene interaction studies suggest that mec-2 positively regulates the activity of the putative mechanosensory transduction channel (and the present paper), comprised in part of proteins encoded by the two degenerin genes mec-4 and mec-10 The central region of the mec-2 protein (MEC-2) is very similar to stomatin, an integral membrane protein (band 7.2b) in human red blood cells that is thought to regulate cation conductance. MEC-2-LacZ fusions are distributed along the touch receptor axons. This axonal distribution, which is mediated by the mec-2-specific amino terminus, is disrupted by mutations in mec-12, an alpha-tubulin gene needed for touch cell function. Our results indicate that MEC-2 links the mechanosensory channel and the microtubule cytoskeleton of the touch receptor neurons. Such linkage provides the basis for a mechanism of mechanosensation whereby microtubule displacement leads to channel opening. PMID- 7477351 TI - Partial activation of CD8+ T cells by a self-derived peptide. AB - T cells are normally activated when the peptide for which they are specific is presented to them in the context of the appropriate major histocompatibility complex (MHC) (class I and Class II for CD8+ and CD4+ T cells, respectively). An increasing body of evidence indicates that structural homologues of the immunogenic peptide can partially activate or antagonize CD4+ T cells. CD8+ T cells may also be partially antagonized by such peptides, and self-derived peptides of this type may play a role in CD8+ T cell selection in the thymus. Activated CD8+ T cells lyse their targets by perforin-dependent granule exocytosis and by inducing apoptosis mediated by CD95 (also known as Fas or APO1) with its ligand (CD95L). Here we show that a clone of Kd-restricted CD8+ T cells specific for influenza haemagglutinin, which can also be activated in a crossreactive manner by a peptide derived from a myeloma tumour immunoglobulin heavy-chain variable region (IgVH) to kill by both routes, kills only by the CD95 CD95L pathway when stimulated by the corresponding germline IgVH peptide. As this germline IgVH peptide differs from the tumour peptide only at a single position buried in the MHC-binding groove, this indicates that CD95-CD95L-mediated killing can be triggered independently of the perforin-mediated pathway, and can be selectively affected by changes in MHC conformation. PMID- 7477352 TI - Perinatal lethality and blocked B-cell development in mice lacking the tyrosine kinase Syk. AB - The tyrosine kinase Syk (relative molecular mass 72,000), which is widely expressed in haematopoietic cells, becomes associated with and activated by engagement of the B-cell antigen receptor. Furthermore, it has been implicated in signalling through the receptors for interleukin-2 (IL-2), granulocyte colony stimulating factor (G-CSF) and Fc, the T cell receptor, as well as through receptors for several platelet agonists. A homologous kinase, ZAP-70, is crucial in signalling through the T-cell receptor and in T-cell development. Using homologous recombination in embryonic stem cells, we created mice null for the syk gene which showed petechiae in utero and died shortly after birth. Irradiated mice reconstituted with Syk-deficient fetal liver showed a block in B-cell development at the pro-B to pre-B cell transition, consistent with a key role for Syk in pre-B-cell receptor signalling. Despite the production of small numbers of immature B cells, Syk-deficient radiation chimaeras failed to accumulate mature B cells, indicating a possible role for this protein in the production or maintenance of mature B cells. In addition, whereas the development of alpha beta T cells proceeded normally, Syk-deficient mice showed impaired development of thymocytes using the V gamma 3 variable region gene (V gamma 3+ thymocytes). Finally, we show that Syk is not required for signalling through the IL-2 and G CSF receptors. PMID- 7477353 TI - Syk tyrosine kinase required for mouse viability and B-cell development. AB - The Syk cytoplasmic protein-tyrosine kinase has two amino-terminal SH2 domains and a carboxy-terminal catalytic domain. Syk, and its close relative ZAP-70, are apparently pivotal in coupling antigen- and Fc-receptors to downstream signalling events. Syk associates with activated Fc receptors, the T cell receptor complex and the B-cell antigen-receptor complex (BCR) in immature and mature B lymphocytes. On receptor activation, the tandem SH2 domains of Syk bind dual phosphotyrosine sites in the conserved ITAM motifs of receptor signalling chains, such as the immunoglobulin alpha and beta-chains of the BCR, leading to Syk activation. Here we have investigated Syk function in vivo by generating a mouse strain with a targeted mutation in the syk gene. Homozygous syk mutants suffered severe haemorrhaging as embryos and died perinatally, indicating that Syk has a critical role in maintaining vascular integrity or in wound healing during embryogenesis. Analysis of syk-/- lymphoid cells showed that the syk mutation impaired the differentiation of B-lineage cells, apparently by disrupting signalling from the pre-BCR complex and thereby preventing the clonal expansion, and further maturation, of pre-B cells. PMID- 7477354 TI - Phosphorylation of Raf by ceramide-activated protein kinase. AB - The sphingomyelin pathway, initiated by hydrolysis of sphingomyelin to ceramide and stimulation of a Ser/Thr ceramide-activated protein (CAP) kinase, mediates tumour necrosis factor-alpha (TNF-alpha) and interleukin-1 beta action. CAP kinase is membrane-bound and proline-directed, recognizing the minimal substrate motif Thr-Leu-Pro. TNF may use the sphingomyelin pathway to signal Raf1 to activate the MAP kinase cascade. Evidence shows that cytoplasmic Raf1 binds to GTP-ras upon cellular stimulation, is recruited to the plasma membrane, and activated. How membrane-bound Raf1 is activated is uncertain, but regulation of its kinase activity may involve its phosphorylation. Specific Raf kinases, however, have not hitherto been identified. Here we report that CAP kinase phosphorylates Raf1 on Thr 269, increasing its activity towards MEK (MAP kinase or ERK kinase). Moreover, in intact HL-60 cells, CAP kinase complexes with Raf1 and, in response to TNF and ceramide analogues, phosphorylates and activates Raf1, implicating CAP kinase as a link between the TNF receptor and Raf1. PMID- 7477355 TI - AIDS vaccine 'needs focused effort' as drug firms back off research. PMID- 7477357 TI - NIH discovers who counts as 'essential'. PMID- 7477358 TI - Biosafety rules will regulate international GMO transfers. PMID- 7477356 TI - Black scientist faces inquiry at South African university. PMID- 7477359 TI - Swiss researchers adopt voluntary rules on exports. PMID- 7477361 TI - Review for radiation effects research. PMID- 7477360 TI - Guidelines may boost gene therapy in Japan. PMID- 7477362 TI - Europe tries again on biotechnology patents. PMID- 7477363 TI - Patterns of pattern formation. PMID- 7477364 TI - Cell-cell signalling. neu tack on neuregulin. PMID- 7477365 TI - Neurobiology. Nitric oxide and bad behaviour. PMID- 7477366 TI - Blood coagulation. The thrombin paradox. PMID- 7477367 TI - Tuning stochastic resonance. PMID- 7477368 TI - Toxic mutants in Charcot's sclerosis. PMID- 7477369 TI - Origin of yaws in the Pleistocene. PMID- 7477370 TI - Noise-induced enhancement of signal transduction across voltage-dependent ion channels. AB - The presence of noise in a signal transduction system usually interferes with its ability to transfer information reliably. But many nonlinear systems can use noise to enhance performance, and this phenomenon, called stochastic resonance, may underlie the extraordinary ability of some biological systems to detect and amplify small signals in noisy environments. Previous work has demonstrated the occurrence of stochastic resonance in a complex system of biological transducers and neural signal pathways, but the possibility that it could occur at the sub cellular level has remained open. Here we report the observation of stochastic resonance in a system of voltage-dependent ion channels formed by the peptide alamethicin. A hundred-fold increase in signal transduction induced by external noise is accompanied by a growth in the output signal-to-noise ratio. The system of ion channels considered here represents the simplest biological system yet known to exhibit stochastic resonance. PMID- 7477371 TI - A recent common ancestry for human Y chromosomes. AB - The male-specific portion of the Y chromosome is especially useful for studies of human origins. Patterns of nucleotide variation that are neutral with respect to fitness should permit estimates of when and where ancestral Y chromosomes existed. However, variation on the human Y chromosome has been observed to be greatly reduced relative to the autosomes and the X chromosome. One explanation is that selection for a favourable mutation on the non-recombining portion of the Y chromosome has resulted in the recent fixation of a single Y haplotype. A 2.6 kilobase fragment encompassing a polymorphic Alu insertion was sequenced from 16 human and four chimpanzee Y chromosomes. Patterns of nucleotide sequence diversity and divergence provide no evidence for a recent, strong selective sweep on the human Y chromosome. The time back to a common ancestral human Y chromosome is estimated to be 188,000 years, with a 95% confidence interval from 51,000 to 411,000 years. These results are consistent with autosomal and mitochondrial DNA studies that suggest a long-term human effective population size of 10,000 and a sex ratio of 1 (ref. 7). These inferences contradict predictions of the multiregional hypothesis positing a widespread transformation of Homo erectus populations into Homo sapiens. PMID- 7477372 TI - Sequence variation of the human Y chromosome. AB - We have generated over 100 kilobases of sequence from the nonrecombining portion of the Y chromosomes from five humans and one common chimpanzee. The human subjects were chosen to match the earliest branches of the human mitochondrial tree. The survey of 18.3 kilobases from each human detected only three sites at which substitutions were present, whereas the human and chimpanzee sequences showed 1.3% divergence. The coalescence time estimated from our Y chromosome sample is more recent than that of the mitochondrial genome. A recent coalescence time for the Y chromosome could have been caused by the selected sweep of an advantageous Y chromosome or extensive migration of human males. PMID- 7477373 TI - Stereoscopic depth perception at high velocities. AB - The view of the world from different perspectives provided by the two eyes is used by the human visual system to compute the relative distances and solid shapes of objects. However, the traditional theory of binocular disparity takes little account of the fact that a moving target will stimulate many different sets of disparate points in the two eyes with a range of temporal delays. Here we show that stereoacuity for periodic grating is not degraded by velocities of up to 640 degrees s-1 provided that they do not move at a greater rate than 30 cycles s-1. The minimum detectable spatial phase difference between the eyes was equivalent to a spatial phase difference of about 5 degrees and an interocular temporal delay as small as 450 microseconds. We suggest that stereopsis for moving targets is accomplished by neurons having a spatial-temporal phase shift in their receptive fields between the eyes. PMID- 7477374 TI - Behavioural abnormalities in male mice lacking neuronal nitric oxide synthase. AB - In addition to its role in blood vessel and macrophage function, nitric oxide (NO) is a neurotransmitter found in high densities in emotion-regulating brain regions. Mice with targeted disruption of neuronal NO synthase (nNOS) display grossly normal appearance, locomotor activity, breeding, long-term potentiation and long-term depression. The nNOS- mice are resistant to neural stroke damage following middle cerebral artery ligation. Although CO2-induced cerebral vasodilatation in wild-type mice is NO-dependent, in nNOS- mice this vasodilation is unaffected by NOS inhibitors. Establishing a behavioural role for NO has, until now, not been feasible, as NOS inhibitor drugs can only be administered acutely and because their pronounced effects on blood pressure and other body functions obfuscate behavioural interpretations. We now report a large increase in aggressive behaviour and excess, inappropriate sexual behaviour in nNOS- mice. PMID- 7477375 TI - Multiple essential functions of neuregulin in development. AB - Neuregulin (also called NDF, heregulin, GGF and ARIA) is a member of the EGF family which induces growth and differentiation of epithelial, glial and muscle cells in culture. The biological effects of the factor are mediated by tyrosine kinase receptors. Neuregulin can bind directly to erbB3 and erbB4 and receptor heterodimerization allows neuregulin-dependent activation of erbB2 (refs 1, 2, 5). A targeted mutation in mice reveals multiple essential roles of neuregulin in development. Here we show that neuregulin -/- embryos die during embryogenesis and display heart malformations. In addition, Schwann cell precursors and cranial ganglia fail to develop normally. The phenotype demonstrates that in vivo neuregulin acts locally and frequently in a paracrine manner. All cell types affected by the mutation express either erbB3 or erbB4, indicating that either of these tyrosine kinase receptors can be a component in recognition and transmission of essential neuregulin signals. PMID- 7477376 TI - Aberrant neural and cardiac development in mice lacking the ErbB4 neuregulin receptor. AB - Various in vitro studies have suggested that ErbB4 (HER4) is a receptor for the neuregulins, a family of closely related proteins implicated as regulators of neural and muscle development, and of the differentiation and oncogenic transformation of mammary epithelia. Here we demonstrate that ErbB4 is an essential in vivo regulator of both cardiac muscle differentiation and axon guidance in the central nervous system (CNS). Mice lacking ErbB4 die during mid embryogenesis from the aborted development of myocardial trabeculae in the heart ventricle. They also display striking alterations in innervation of the hindbrain in the CNS that are consistent with the restricted expression of the ErbB4 gene in rhombomeres 3 and 5. Similarities in the cardiac phenotype of ErbB4 and neuregulin gene mutants suggest that ErbB4 functions as a neuregulin receptor in the heart; however, differences in the hindbrain phenotypes of these mutants are consistent with the action of a new ErbB4 ligand in the CNS. PMID- 7477377 TI - Requirement for neuregulin receptor erbB2 in neural and cardiac development. AB - The receptor erbB2/neu is a member of the epidermal growth factor receptor (EGFR or erbB) family that also includes erbB3 and erbB4. Amplification of the erbB2/neu gene is found in many cancer types and its overexpression is correlated with a poor prognosis for breast and ovarian cancer patients. Investigation of the biology of erbB2 led to the identification of a family of ligands termed neuregulins which included the neu-differentiation factors, the heregulins, a ligand with acetylcholine-receptor-inducing activity and glial growth factor. Several lines of evidence suggest that heterodimerization of erbB2 with other erbB receptors is required for neuregulin signalling. Here we investigate the developmental role of erbB2 in mammalian development in mice carrying an erbB2 null allele. We find that mutant embryos die before E11, probably as a result of dysfunctions associated with a lack of cardiac trabeculae. Development of cranial neural-crest-derived sensory ganglia was markedly affected. DiI retrograde tracing revealed that the development of motor nerves was also compromised. Our results demonstrate the importance of erbB2 in neural and cardiac development. PMID- 7477378 TI - A huntingtin-associated protein enriched in brain with implications for pathology. AB - Huntington's disease (HD) is an autosomal dominant neurodegenerative disorder caused by an expanding polyglutamine repeat in the IT15 or huntingtin gene. Although this gene is widely expressed and is required for normal development, the pathology of HD is restricted to the brain, for reasons that remain poorly understood. The huntingtin gene product is expressed at similar levels in patients and controls, and the genetics of the disorder suggest that the expansion of the polyglutamine repeat induces a toxic gain of function, perhaps through interactions with other cellular proteins. Here we report the identification of a protein (huntingtin-associated protein (HAP)-1) that binds to huntingtin. This binding is enhanced by an expanded polyglutamine repeat, the length of which is also known to correlate with the age of disease onset. The HAP 1 protein is enriched in the brain, suggesting a possible basis for the selective brain pathology of HD. PMID- 7477379 TI - Polyglutamine expansion as a pathological epitope in Huntington's disease and four dominant cerebellar ataxias. AB - A polyglutamine expansion (encoded by a CAG repeat) in specific proteins causes neurodegeneration in Huntington's disease (HD) and four other disorders, by an unknown mechanism thought to involve gain of function or toxicity of the mutated protein. The pathological threshold is 37-40 glutamines in three of these diseases, whereas the corresponding normal proteins contain polymorphic repeats of up to about 35 glutamines. The age of onset of clinical manifestations is inversely correlated to the length of the polyglutamine expansion. Here we report the characterization of a monoclonal antibody that selectively recognizes polyglutamine expansion in the proteins implicated in HD and in spinocerebellar ataxia (SCA) 1 and 3. The intensity of signal depends on the length of the polyglutamine expansion, and the antibody also detects specific pathological proteins expected to contain such expansion, in SCA2 and in autosomal dominant cerebellar ataxia with retinal degeneration, whose genes have not yet been identified. PMID- 7477380 TI - Renal abnormalities and an altered inflammatory response in mice lacking cyclooxygenase II. AB - Prostaglandins have wide-ranging effects in the body and are thought to be important mediators of inflammation. Cyclooxygenase (COX) plays a key regulatory role in prostaglandin synthesis, and occurs in both constitutive (COX-1) and inducible (COX-2) isoforms. COX-1 is thought to provide cytoprotective effects, whereas COX-2 is both inducible and the major isoform of inflammatory cells. Reduction of prostaglandin production by inhibition of cyclooxygenases appears to be the main mechanism of action of most non-steroidal anti-inflammatory drugs (NSAIDS). Here we present an animal model of COX-2 deficiency that was generated by gene targeting. Defects in null mice correlating with reduced viability included renal alterations, characteristic of renal dysplasia (100% penetrance), and cardiac fibrosis (50% penetrance). Female Cox-2-/- mice were infertile. COX-2 deficiency failed to alter inflammatory responses in several standard models, but striking mitigation of endotoxin-induced hepatocellular cytotoxicity was observed. PMID- 7477381 TI - Involvement of Ral GTPase in v-Src-induced phospholipase D activation. AB - An early response to the tyrosine kinase activity of v-Src is an increase in phospholipase D (PLD) activity, which leads to the generation of biologically active lipid second messengers, including phosphatidic acid, lysophosphatidic acid and diacylglycerol. We have recently demonstrated that v-Src-induced PLD activity is mediated by Ras, although Ras involvement was indirect, requiring a cytosolic factor for PLD activation. Ras interacts with and activates Ral-GDS, the exchange factor responsible for the activation of Ral GTPases. Here we report that this newly identified Ras/Ral signalling pathway mediates PLD activation by v-Src. PLD activity could be precipitated from v-Src-transformed cell lysates with immobilized RalA protein and with an anti-Ral antibody. A mutation to the region of RalA analogous to the 'effector domain' of Ras did not reduce the ability of RalA to complex with PLD, although deletion of a Ral-specific amino terminal region did. Overexpression of RalA potentiated PLD activation by v-Src, and expression of dominant negative RalA mutants inhibited both v-Src- and v-Ras induced PLD activity. Thus RalA is involved in the tyrosine kinase activation of PLD through its unique N terminus, and that PLD is a downstream target of a Ras/Ral GTPase cascade. PMID- 7477382 TI - Conversion of thrombin into an anticoagulant by protein engineering. AB - At sites of vascular injury, thrombin interacts with multiple procoagulant substrates, to mediate both fibrin clotting and platelet aggregation. But upon binding to thrombomodulin on the vascular endothelium, thrombin instead activates protein C, thereby functioning as an anticoagulant and attenuating clot formation. Upon infusion in vivo, both the procoagulant and anticoagulant effects of thrombin were observed. Preliminary studies indicating that thrombin's protein C activating and fibrinogen clotting activities could be dissociated by mutagenesis suggested to us that a thrombin variant that lacked procoagulant activity while retaining anticoagulant function might be an attractive antithrombotic agent. Using protein engineering, we introduced a single substitution, E229A, that substantially shifted thrombin's specificity in favour of the anticoagulant substrate, protein C. In monkeys, this modified thrombin functioned as an endogenous protein C activator demonstrating dose-dependent, reversible anticoagulation without any indication of procoagulant activity. Notably, template bleeding times were not prolonged, suggesting a reduced potential for bleeding complications. PMID- 7477383 TI - A protein catalytic framework with an N-terminal nucleophile is capable of self activation. AB - The crystal structures of three amidohydrolases have been determined recently: glutamine PRPP amidotransferase (GAT), penicillin acylase, and the proteasome. These enzymes use the side chain of the amino-terminal residue, incorporated in a beta-sheet, as the nucleophile in the catalytic attack at the carbonyl carbon. The nucleophile is cysteine in GAT, serine in penicillin acylase, and threonine in the proteasome. Here we show that all three enzymes share an unusual fold in which the nucleophile and other catalytic groups occupy equivalent sites. This fold provides both the capacity for nucleophilic attack and the possibility of autocatalytic processing. We suggest the name Ntn (N-terminal nucleophile) hydrolases for this structural superfamily of enzymes which appear to be evolutionarily related but which have diverged beyond any recognizable sequence similarity. PMID- 7477384 TI - Regulation of neural induction by the Chd and Bmp-4 antagonistic patterning signals in Xenopus. PMID- 7477385 TI - Go fish for patents. PMID- 7477387 TI - UK court rejects appeal over hepatitis C kits. PMID- 7477386 TI - Open access to sequence data 'will boost hunt for breast cancer gene'. PMID- 7477388 TI - Oncomouse hearing ends up in confusion. PMID- 7477389 TI - Call for more research into health effects of Chernobyl accident. PMID- 7477390 TI - Deaths prompt calls for drug control review. PMID- 7477391 TI - Quis custodiet ipsos custodes. PMID- 7477392 TI - Quis custodiet ipsos custodes? PMID- 7477393 TI - Aversion to xenotransplantation. PMID- 7477394 TI - Aversion to xenotransplantation. PMID- 7477395 TI - Aversion to xenotransplantation. PMID- 7477396 TI - The prevalent distrust of science. AB - An account of the now common distrust of science, an explanation of its origins and a suggestion of what may, and indeed, should be done about it. PMID- 7477397 TI - Signal transduction. Beating a path to Myc. PMID- 7477398 TI - Spinal injury. Overcoming inhibition. PMID- 7477399 TI - Ion channels. Annexins taken to task. PMID- 7477400 TI - The structure of an intermediate in class II MHC maturation: CLIP bound to HLA DR3. AB - A complex between HLA-DR3 and a fragment of invariant chain called CLIP was isolated from a human cell line defective in antigen presentation and its X-ray crystal structure determined. Previous data indicate that this complex is an intermediate in class II histocompatibility maturation, occurring between invariant chain-DR3 and antigenic peptide-DR3 complexes. The structure shows that the CLIP fragment binds to DR3 in a way almost identical to that in which antigenic peptides bind class II histocompatibility glycoproteins. The structure is the substrate for the loading of antigenic peptides by an exchange process catalysed by DM. PMID- 7477401 TI - Control of protein-ligand recognition using a stimuli-responsive polymer. AB - Stimuli-responsive polymers exhibit reversible phase changes in response to changes in environmental factors such as pH or temperature. Conjugating such polymers to antibodies and proteins provides molecular systems for applications such as affinity separations, immunoassays and enzyme recovery and recycling. Here we show that conjugating a temperature-sensitive polymer to a genetically engineered site on a protein allows the protein's ligand binding affinity to be controlled. We synthesized a mutant of the protein streptavidin to enable site specific conjugation of the responsive polymer near the protein's binding site. Normal binding of biotin to the modified protein occurs below 32 degrees C, whereas above this temperature the polymer collapses and blocks binding. The collapse of the polymer and thus the enabling and disabling of binding, is reversible. Such environmentally triggered control of binding may find many applications in biotechnology and biomedicine, such as the control of enzyme reaction rates and of biosensor activity, and the controlled release of drugs. PMID- 7477402 TI - A plausibly prebiotic synthesis of phosphonic acids. AB - The insolubility of calcium phosphate in water is a significant stumbling block in the chemistry required for the origin of life. The discovery of alkyl phosphonic acids in the Murchison meteorite suggests the possibility of delivery of these water-soluble, phosphorus-containing molecules by meteorites or comets to the early Earth. This could have provided a supply of organic phosphorus for the earliest stages of chemical evolution; although probably not components of early genetic systems, phosphonic acids may have been precursors to the first nucleic acids. Here we report the synthesis of several phosphonic acids, including the most abundant found in the Murchison meteorite, by ultraviolet irradiation of orthophosphorous acid in the presence of formaldehyde, primary alcohols, or acetone. We argue that similar reactions might explain the presence of phosphonic acids in Murchison, and could also have occurred on the prebiotic Earth. PMID- 7477403 TI - Insertions and duplications of mtDNA in the nuclear genomes of Old World monkeys and hominoids. AB - Using oligonucleotide primers designed to match conserved regions of mammalian mitochondrial DNA (mtDNA), we have amplified and sequenced two divergent cytochrome b nuclear pseudogenes from orangutan cellular DNA. Evolutionary analysis suggests that a nuclear transfer occurred about 30 million years ago on the lineage leading to the catarrhines (Old World monkeys and hominoids), and involved a long (at least 3 kilobases), probably damaged, piece of mtDNA. After this transfer, the pseudogene duplicated, giving rise to the two copies that are probably present in all hominoids, including humans. More recent transfers involving the entire cytochrome b gene have also occurred in the Old World monkeys. Such nuclear copies of mtDNA can confound phylogenetic and population genetic studies, and be an insidious source of DNA contamination of 'ancient' and forensic DNA. Indeed, contamination with these anciently transferred human pseudogenes is almost certainly the source of the cytochrome b sequences recently reported from 'dinosaur bone DNA'. PMID- 7477404 TI - A nuclear 'fossil' of the mitochondrial D-loop and the origin of modern humans. AB - Mammalian mitochondrial DNA sequences evolve more rapidly than nuclear sequences. Although the rapid rate of evolution is an advantage for the study of closely related species and populations, it presents a problem in situations where related species, used as outgroups in phylogenetic analyses, have accumulated so much change that multiple substitutions obliterate the phylogenetic information. However, mitochondrial DNA sequences are frequently inserted into the nuclear genome, where they presumably evolve as nuclear pseudogene sequences and therefore more slowly than their mitochondrial counterparts. Such sequences thus represent molecular 'fossils' that could shed light on the evolution of the mitochondrial genome and could be used as outgroups in situations where no appropriate outgroup species exist. Here we show that human chromosome 11 carries a recent integration of the mitochondrial control region that can be used to gain further insight into the origin of the human mitochondrial gene pool. PMID- 7477405 TI - Visual cortical mechanisms detecting focal orientation discontinuities. AB - Neurons in the primary visual cortex (V1) respond in well defined ways to stimuli within their classical receptive field, but these responses can be modified by stimuli overlying the surrounding area. For example patch-suppressed cells respond to gratings of a specific orientation within their classical receptive field, but the response diminishes if the grating is expanded to cover the surrounding area. We report here more complex effects in many such cells. When stimulated at their optimal orientation, introducing a surrounding field at a significantly different (for example, orthogonal) orientation enhanced their output by both a disinhibitory mechanism and an active facilitatory mechanism producing 'supra-optimal' responses. Importantly, some cells responded well if the orientations of centre and surround stimuli were swapped. The output reflected the discontinuity because neither stimulus component alone was effective. Under these stimulus conditions simultaneously recorded cells with orthogonally oriented receptive fields showed correlated firing consistent with neuronal binding to the configuration. We propose a mechanism integrating orientation-dependent information over adjacent areas of visual space to represent focal orientation discontinuities such as junctions or corners. PMID- 7477406 TI - Topographical representations of mental images in primary visual cortex. AB - We report here the use of positron emission tomography (PET) to reveal that the primary visual cortex is activated when subjects close their eyes and visualize objects. The size of the image is systematically related to the location of maximal activity, which is as expected because the earliest visual areas are spatially organized. These results were only evident, however, when imagery conditions were compared to a non-imagery baseline in which the same auditory cues were presented (and hence the stimuli were controlled); when a resting baseline was used (and hence brain activation was uncontrolled), imagery activation was obscured because of activation in visual cortex during the baseline condition. These findings resolve a debate in the literature about whether imagery activates early visual cortex and indicate that visual mental imagery involves 'depictive' representations, not solely language-like descriptions. Moreover, the fact that stored visual information can affect processing in even the earliest visual areas suggests that knowledge can fundamentally bias what one sees. PMID- 7477407 TI - Recovery from spinal cord injury mediated by antibodies to neurite growth inhibitors. AB - There is little axonal growth after central nervous system (CNS) injury in adult mammals. The administration of antibodies (IN-1) to neutralize the myelin associated neurite growth inhibitory proteins leads to long-distance regrowth of a proportion of CNS axons after injury. Our aim was: to determine if spinal cord lesion in adult rats, followed by treatment with antibodies to neurite growth inhibitors, can lead to regeneration and anatomical plasticity of other spinally projecting pathways; to determine if the anatomical projections persist at long survival intervals; and to determine whether this fibre growth is associated with recovery of function. We report here that brain stem-spinal as well as corticospinal axons undergo regeneration and anatomical plasticity after application of IN-1 antibodies. There is a recovery of specific reflex and locomotor functions after spinal cord injury in these adult rats. Removal of the sensorimotor cortex in IN-1-treated rats 2-3 months later abolished the recovered contact-placing responses, suggesting that the recovery was dependent upon the regrowth of these pathways. PMID- 7477408 TI - A switch between two modes of synaptic transmission mediated by presynaptic inhibition. AB - Presynaptic inhibition reduces chemical synaptic transmission in the central nervous system between pairs of neurons, but its role(s) in shaping the multisynaptic interactions underlying neural network activity are not well studied. We therefore used the crustacean stomatogastric nervous system to study how presynaptic inhibition of the identified projection neuron, modulatory commissural neuron 1 (MCN1), influences the MCN1 synaptic effects on the gastric mill neural network. Tonic MCN1 discharge excites gastric mill network neurons and activates the gastric mill rhythm. One network neuron, the lateral gastric (LG) neuron, presynaptically inhibits MCN1 and is electrically coupled to its terminals. We show here that this presynaptic inhibition selectively reduces or eliminates transmitter-mediated excitation from MCN1 without reducing its electrically mediated excitatory effects, thereby switching the network neurons excited by MCN1. By switching the type of synaptic output from MCN1 and, hence, the activated network neurons, this presynaptic inhibition is pivotal to motor pattern generation. PMID- 7477409 TI - Altered Hox expression and segmental identity in Mll-mutant mice. AB - The mixed-lineage leukaemia gene (MLL/HRX/ALL-1) is disrupted by chromosomal translocation in human acute leukaemias that often display mixed lymphoid-myeloid phenotypes and present in infancy. MLL possesses a highly conserved SET domain also found in Drosophila trithorax (trx) and Polycomb group (Pc-G) genes, which are known to regulate homeotic genes (HOM-C) in a positive or negative fashion, respectively. Mll was targeted in mice by homologous recombination in embryonic stem (ES) cells to assess its role in pattern development. Mll heterozygous (+/-) mice had retarded growth, displayed haematopoietic abnormalities, and demonstrated bidirectional homeotic transformations of the axial skeleton as well as sternal malformations. Mll deficiency (-/-) was embryonic lethal. Anterior boundaries of Hoxa-7 and Hoxc-9 expression were shifted posteriorly in Mll +/- embryos, but their expression was abolished in Mll -/- embryos. Thus Mll is required for proper segment identity in mammals, displays haplo-insufficiency, and positively regulates Hox gene expression. PMID- 7477410 TI - Myc but not Fos rescue of PDGF signalling block caused by kinase-inactive Src. AB - Growth factors such as platelet-derived growth factor (PDGF) elicit the transcriptional activation of a large number of immediate early genes (many of which encode transcription factors), and ultimately DNA synthesis. Both AP1 and Myc are activated in fibroblasts in response to growth factor stimulation, and various experiments suggest their importance in proliferation. Src family kinases are required for PDGF (and other growth factors) to induce DNA synthesis. We have examined which transcription factors, when constitutively expressed, 'rescue' the block elicited by dominant negative Src. We report here that Myc, but not Fos and/or Jun, was able to rescue the block. In contrast, Fos and Jun, but not Myc, rescued the block induced by dominant negative Ras. Our data suggest that Src kinases control the transcriptional activation of Myc. PMID- 7477411 TI - Crystal structure of the annexin XII hexamer and implications for bilayer insertion. AB - Annexins are a family of calcium- and phospholipid-binding proteins implicated in a number of biological processes including membrane fusion and ion channel formation. The crystal structure of the annexin XII hexamer, refined at 2.8 A resolution, forms a concave disk with 3-2 symmetry, about 100 A in diameter and 70 A thick with a central hydrophilic pore. Six intermolecular Ca2+ ions are involved in hexamer formation. An additional 18 Ca2+ ions are located on the perimeter of the disk, accessible only from the side of the hexameric disk. On the basis of the hexamer structure we propose here a new mode of protein phospholipid bilayer interaction that is distinct from the hydrophobic insertion of typical membrane proteins. This speculative model postulates the Ca(2+) dependent insertion of the hydrophilic annexin XII hexamer into phospholipid bilayers with local reorientation of the bilayer phospholipids. PMID- 7477412 TI - Mapping the genome one molecule at a time--optical mapping. PMID- 7477413 TI - [Glycolipids of the cell surface--biochemistry of their decomposition]. AB - Glycosphingolipids (GSL) form cell-type-specific patterns on the surface of eukaryotic cells. For the maintenance of these patterns, biosynthesis, intracellular transport, and degradation of GSL are thought to proceed in a coordinated fashion. After transport from the plasma membrane to the lysosomes the degradation of GSL requires protein cofactors to solubilize the membrane bound substrates of the degradative enzymes. Inborn errors of metabolism leading to defective hydrolases or activator proteins give rise to sphingolipid storage diseases; in some cases the residual enzyme activity can be correlated with the clinical manifestations. PMID- 7477414 TI - Unexpectedly slow decrease of Chernobyl-derived radiocesium in air and deposition in Bavaria/Germany. PMID- 7477415 TI - Evidence of contamination in PCR laboratory disposables. PMID- 7477416 TI - Vision through colored eyes. PMID- 7477418 TI - Smoking cessation and nicotine substitution modulate eicosanoid synthesis ex vivo in man. AB - The effects of smoking cessation with and without nicotine substitution on prostaglandin E2, leukotriene B4, leukotriene E4, and thromboxane B2 synthesis ex vivo in man were investigated. Blood samples were obtained from 20 healthy non smoking controls and from 30 healthy smoking volunteers before and 3, 7 and 14 days after smoking cessation. Half of the smokers used nicotine chewing gum as a substitution therapy. Urinary cotinine and trans-3'-hydroxycotinine as well as thiocyanate excretions were used as indicators for the use of nicotine chewing gum and smoking, respectively. Prostaglandin E2, leukotriene E4, and thromboxane B2 were measured from whole blood after calcium ionophore A23187 stimulation by direct radioimmunoassay and leukotriene B4 by RP-HPLC. Prostaglandin E2 and thromboxane B2 syntheses were about three times and leukotriene B4 and E4 syntheses four times higher in smokers than in controls. Three days after smoking cessation without nicotine substitution, levels were lowered significantly to about 70%, 80%, 45% and 60% of the initial values; and after 14 days to 55%, 80%, 45% and 50%, respectively. In the nicotine substitution group no significant decreases were seen during the two-week follow-up. The increased level of eicosanoid synthesis detected in smokers in this ex vivo study may contribute to the harmful cardiovascular effects of smoking. Long-term nicotine substitution might diminish the beneficial effects of smoking cessation due to the possible stimulatory effects of nicotine and cotinine on eicosanoid synthesis even in vivo. PMID- 7477417 TI - Classification of alpha 1-adrenoceptor subtypes. AB - Two alpha 1-adrenoceptor subtypes (alpha 1A and alpha 1B) have been detected in various tissues by pharmacological techniques, and three distinct cDNAs encoding alpha 1-adrenoceptor subtypes have been cloned. The profile of an increasing number of subtype-selective compounds at cloned and endogenous receptors recently has facilitated alignment between cloned and pharmacologically defined alpha 1 adrenoceptor subtypes. Thus, alpha 1a-adrenoceptors (previously designated alpha 1c), alpha 1b-adrenoceptors and alpha 1d-adrenoceptors (previously designated alpha 1a, alpha 1d or alpha 1a/d) are now recognized. Since the alpha 1d adrenoceptor shares characteristics with both alpha 1A- and alpha 1B adrenoceptors, tissues previously reported to express alpha 1A- and/or alpha 1B adrenoceptors may additionally contain alpha 1d-adrenoceptors. This article reviews the features of all three subtypes and discusses possible pitfalls in their pharmacological identification. PMID- 7477419 TI - Mechanisms of central endothelin-induced hypotension. AB - The aims of the present study were i) to determine the type of endothelin receptor(s) mediating the hypotension produced by central administration of endothelin-1 (ET-1), ii) to delineate the hemodynamic factors contributing to this hypotension and iii) to differentiate between the neural and cerebrovascular actions of ET-1. Towards these objectives, we monitoreal blood flow from the choroid plexus of the IVth cerebral ventricle (4CV) as an index of local cerebral blood flow (CBF); also, aortic blood flow (ABF) and cutaneous microvascular blood flow (CMF) of the hindpaw were monitored. In anesthetized, ventilated rats, ET-1 (1, 3 and 10 pmol) applied to the 4CV produced significant decreases in mean arterial blood pressure (15 +/- 4%, 34 +/- 3% and 37 +/- 3% respectively); hypotension was sustained at the two higher doses. ET-1 also produced a profound and sustained reduction in CBF (36 +/- 10%, 54 +/- 10% and 57 +/- 11% respectively). Prior administration of a low dose (1 nmol) of the ETA receptor selective antagonist, BQ-123 [cyclo (D-Trp-D-Asp-L-Pro-D-Val-L-Leu)], abolished only the central ET-1-induced hypotension; the decreases in CBF were not altered (57 +/- 11% and 56 +/- 6% respectively after 3 and 10 pmol). Pretreatment with a high dose (20 nmol) of BQ-123 attenuated but did not abolish the CBF response to 10 pmol of ET-1 (-26 +/- 1% vs. -57 +/- 11%).(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7477420 TI - Strychnine affects catecholamine secretion from bovine adrenal medulla chromaffin cells. AB - The effect of strychnine on evoked release of catecholamines from a primary culture of bovine adrenal medullary cells was investigated. Strychnine at > 1 microM inhibited catecholamine release stimulated by 10 microM acetylcholine, or 10 microM nicotine, but not by excess K+ (59 mM), the sodium ionophore veratridine (100 microM) or the calcium ionophore A-23187 (10 microM). The inhibitory response elicited by exposure of the cells to strychnine was rapid (< 3 min) and competitive with acetylcholine. High concentrations of acetylcholine (1 mM) completely overcame this inhibition. Strychnine might be acting on a regulatory site of the nicotinic-cholinergic receptor, which is genetically similar to the strychnine-binding 48 KD subunit of the glycine receptor. PMID- 7477421 TI - Cultured chick sympathetic neurons: modulation of electrically evoked noradrenaline release by P2-purinoceptors. AB - The present study investigates the pharmacological profile of P2-purinoceptors modulating noradrenaline release from cultured chick sympathetic neurons. ATP (30 microM-3 mM) and 2-methylthio-ATP (3-100 microM), but not alpha, beta-methylene ATP (up to 100 microM), caused a significant facilitation of electrically evoked [3H]-noradrenaline release when added 2 min before depolarization. The facilitation declined with time of exposure suggesting receptor desensitization. The facilitatory effect was markedly diminished by the P2-purinoceptor antagonists reactive blue 2 (3 microM) and suramin (300 microM), but not changed by mecamylamine (10 microM), a nicotinic receptor antagonist. At 1 mM and higher concentrations, ATP added for 12 min, inhibited noradrenaline release; release was virtually abolished by 6 mM ATP. The inhibitory effect of ATP was slightly diminished by suramin but not affected by reactive blue 2. Electrically evoked [3H]-noradrenaline release remained unaffected in the presence of the adenosine (P1)-receptor agonists R(-)N6-(2-phenylisopropyl)adenosine (R-PIA), 2-[p-(2 carboxyethyl) phenylethylamino]-5'-N-ethylcarboxamidoadenosine (NECA), and N6-2 (4-aminophenyl)ethyladenosine (APNEA), used up to 1 microM. The present results confirm the existence of two P2-purinoceptors affecting noradrenaline release: 1) a facilitatory receptor which is activated by 2-methylthio-ATP as well as ATP, and blocked by suramin as well as reactive blue 2, and 2) an inhibitory receptor which is activated by ATP, only slightly affected by suramin but not at all by reactive blue 2 and does not belong to the established P2-purinoceptor subtypes. PMID- 7477422 TI - Cultured chick sympathetic neurons: ATP-induced noradrenaline release and its blockade by nicotinic receptor antagonists. AB - The ATP-induced increase in tritium outflow from cultured chick sympathetic neurons prelabelled with [3H]-noradrenaline was investigated. Seven days-old dissociated cell cultures of embryonic paravertebral ganglia, loaded with [3H] noradrenaline (0.05 microM), were superfused in the presence of (+)-oxaprotiline and exposed to ATP, ATP-analogues, or 1,1-dimethyl-4-piperazinium (DMPP) for 2 min. ATP (3 microM-3 mM), 2-methylthio-ATP (3-100 microM), as well as DMPP (10 and 100 microM) induced a significant overflow of tritium. The EC50-value of ATP was 20 microM. Both the ATP-induced and the DMPP-induced tritium overflow was Ca(2+)-dependent and sensitive to tetrodotoxin (0.3 microM) and omega-conotoxin (0.1 microM); in addition, it was inhibited by the alpha 2-adrenoceptor agonist 5 bromo-6-(2-imidazoline-2-ylamino)-quinoxaline (UK-14,304; 1 microM). The effects of ATP and DMPP were not additive. The ATP-induced as well as the DMPP-induced overflow of tritium was diminished by the P2-purinoceptor antagonists suramin (300 microM) and reactive blue 2 (3 microM); in all 4 cases, the inhibition amouted to approximately 40%. The tritium overflow induced by ATP or DMPP was almost abolished by the nicotinic receptor antagonist mecamylamine (10 microM) and markedly inhibited by hexamethonium (100 microM). Neither ATP nor electrical stimulation caused an overflow of tritium from cultures loaded with [3H]-choline. The results suggest that ATP at mumolar concentrations induces noradrenaline release from cultured chick sympathetic neurons via an action on a subclass of the nicotinic cholinoceptor. PMID- 7477423 TI - Subclassification of presynaptic alpha 2-adrenoceptors: alpha 2A-autoreceptors in rabbit atria and kidney. AB - The study was devised to classify, by means of antagonist affinities, the presynaptic alpha 2-autoreceptors of rabbit atria and kidney in terms of alpha 2A, alpha 2B, alpha 2C and alpha 2D. A set of antagonists was chosen that was able to discriminate between the four subtypes. Small pieces of the left atrium and slices of the kidney cortex were preincubated with 3H-noradrenaline and then superfused and stimulated electrically. In one series of experiments, tissue pieces were stimulated by relatively long pulse trains (1 or 2 min) leading to alpha 2-autoinhibition. All 11 (atria) or 10 (kidney) antagonists increased the evoked overflow of tritium. pEC30% values (concentrations causing 30% increase) were interpolated from concentration-response curves. In a second series of experiments, tissue pieces were stimulated by brief pulse trains (0.4 s) that did not lead to alpha 2-autoinhibition, and concentration-inhibition curves of the alpha 2-selective agonist 5-bromo-6-(2-imidazolin-2-ylamino)-quinoxaline (UK 14,304) were determined. Most of the 11 (atria) or 8 (kidney) antagonists shifted the concentration-inhibition curve of UK 14,304 to the right. pKd values of the antagonists were calculated from the shifts. pEC30% values correlated with pKd values, both in atria (r = 0.728) and in the kidney (r = 0.930). pEC30% values in atria correlated with pEC30% values in the kidney (r = 0.988) and pKd values in atria correlated with pKd values in kidney (r = 0.923). It is concluded that the alpha 2-autoreceptors in atria and the kidney are the same. Comparison with antagonist affinities for prototypic native alpha 2 binding sites, alpha 2 binding sites in cells transfected with alpha 2 subtype genes, and previously classified presynaptic alpha 2-adrenoceptors--all taken from the literature- indicates that both autoreceptors are alpha 2A. This conclusion is reached with either of the two independent estimates of autoreceptor affinity, pEC30% and pKd. The results are compatible with the hypothesis that at least the majority of alpha 2-autoreceptors belong to the alpha 2A/D branch of the alpha 2-adrenoceptor tree, across mammalian or at least rodent and lagomorph species. PMID- 7477424 TI - Subclassification of presynaptic alpha 2-adrenoceptors: alpha 2D-autoreceptors in mouse brain. AB - The study was devised to classify, by means of antagonist affinities, the presynaptic alpha 2-autoreceptors in mouse cerebral cortex in terms of alpha 2A, alpha 2B, alpha 2C and alpha 2D. A set of antagonists was chosen that was able to discriminate between the four subtypes. Slices of the cortex were preincubated with 3H-noradrenaline and then superfused and stimulated electrically. The stimulation periods used (4 pulses, 100 Hz) did not lead to alpha 2 autoinhibition as shown by the lack of an increase by rauwolscine of the evoked overflow of tritium. The alpha 2-selective agonists 5-bromo-6-(2-imidazolin-2 ylamino)-quinoxaline (UK 14,304) and alpha-methylnoradrenaline reduced the evoked overflow. All 10 antagonists shifted the concentration-inhibition curve of UK 14,304 to the right. Rauwolscine also shifted the concentration-inhibition curve of alpha-methylnoradrenaline to the right. pKd values of the antagonists were calculated from the shifts. The pKd values of rauwolscine against UK 14,304 and alpha-methylnoradrenaline were very similar (8.0 and 7.9, respectively). Comparison with antagonist affinities for prototypic native alpha 2 binding sites, alpha 2 binding sites in cells transfected with alpha 2 subtype genes, and previously classified presynaptic alpha 2-adrenoceptors--all taken from the literature--indicates that the alpha 2-autoreceptors in mouse brain cortex are alpha 2D. This is the first subtype determination of alpha 2-autoreceptors in the mouse. It supports the hypothesis that at least the majority of alpha 2 autoreceptors belong to the alpha 2A/D branch of the alpha 2-adrenoceptor tree. PMID- 7477425 TI - Subclassification of presynaptic alpha 2-adrenoceptors: alpha 2D-autoreceptors in guinea-pig atria and brain. AB - The study was devised to classify, by means of antagonist and agonist affinities, the presynaptic alpha 2-autoreceptors in guinea-pig heart atria and brain cortex in terms of alpha 2A, alpha 2B, alpha 2C and alpha 2D. A set of antagonists and agonists was chosen that was able to discriminate between the four subtypes. Small pieces of the atria and slices of the brain cortex were preincubated with 3H-noradrenaline and then superfused and stimulated electrically. In one series of experiments (atria only), tissue pieces were stimulated by relatively long pulse trains (1 min) leading to marked alpha 2-autoinhibition. All 10 antagonists increased the evoked overflow of tritium. pEC30% values (concentrations causing 30% increase) were interpolated from concentration-response curves. In a second series of experiments (atria and brain slices), tissue pieces were stimulated by brief pulse trains (0.4 s or 40 ms) that led to little (atria) or no (brain slices) alpha 2-autoinhibition, and antagonist effects against the alpha 2 selective agonist 5-bromo-6-(2-imidazolin-2- ylamino)-quinoxaline (UK 14,304) were examined. All 10 (atria) or 8 (brain) antagonists shifted the concentration inhibition curve of UK 14,304 to the right. pKd values of the antagonists were calculated from the shifts. In a third series of experiments (brain slices only), also with brief pulse trains (40 ms), pKa values (negative logarithms of dissociation constants of agonist-alpha 2-adrenoceptor complexes) were determined by comparison of concentration-inhibition curves of UK 14,304, guanoxabenz and oxymetazoline in normal tissue and in tissue in which a fraction of the receptors had been blocked by phenoxybenzamine.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7477426 TI - Subclassification of presynaptic alpha 2-adrenoceptors: alpha 2D-autoreceptors and alpha 2D-adrenoceptors modulating release of acetylcholine in guinea-pig ileum. AB - The study was designed to classify in terms of alpha 2A, alpha 2B, alpha 2C and alpha 2D the presynaptic alpha 2-autoreceptors, as well as the alpha 2-receptors modulating the release of acetylcholine, in the myenteric plexus-longitudinal muscle (MPLM) preparation of the guinea-pig ileum. A set of antagonists was chosen that was able to discriminate between the four subtypes. Small pieces of the MPLM preparation were preincubated with 3H-noradrenaline or 3H-choline and then superfused and stimulated electrically. The stimulation periods used (3H noradrenaline: 3 trains of 20 pulses, 50 Hz, train interval 60 s; 3H-choline: single trains of 30 pulses, 0.2 Hz) did not lead to alpha 2-autoinhibition or inhibition of 3H-acetylcholine release by endogenous noradrenaline. The alpha 2 selective agonist 5-bromo-6-(2-imidazolin-2-ylamino)-quinoxaline (UK 14,304) reduced the evoked overflow of tritium in both 3H-noradrenaline and 3H-choline experiments. Most (3H-noradrenaline) or all (3H-choline) of the 10 antagonists shifted the concentration-inhibition curves of UK 14,304 to the right. pKd values of the antagonists were calculated from the shifts. pKd values from 3H noradrenaline experiments correlated with pKd values from 3H-choline experiments (r = 0.981). It is concluded that alpha 2-autoreceptors and alpha 2 heteroreceptors modulating the release of acetylcholine in the MPLM preparation are of the same subtype. Comparison with antagonist affinities for prototypic native alpha 2 binding sites, binding sites in cells transfected with alpha 2 subtype genes, and previously classified presynaptic alpha 2-adrenoceptors--all taken from the literature--indicates that both are alpha 2D.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7477427 TI - Nitric oxide modulates the release of acetylcholine in the ventral striatum of the freely moving rat. AB - The influence of nitric oxide on acetylcholine release in the ventral striatum was investigated by the push-pull superfusion technique in the conscious, freely moving rat. Superfusion with the nitric oxide donors S-nitroso-N acetylpenicillamine or with 3-morpholino-sydnonimine caused a pronounced increase in striatal acetylcholine release. This effect was prevented by superfusion with tetrodotoxin. Pre-superfusion with the guanylyl cyclase inhibitor methylene blue abolished the effect of 3-morpholino-sydnonimine. Superfusion of the ventral striatum with the guanylyl cyclase inhibitor LY83583 decreased acetylcholine release by 60% of basal release, whereas the less specific guanylyl cyclase inhibitor methylene blue was ineffective in this respect. Superfusion of the ventral striatum with inhibitors of nitric oxide synthase also led to different effects on basal acetylcholine release. Superfusion with L-NG-methylarginine did not influence basal acetylcholine release, whereas superfusion with L-NG nitroarginine or with L-NG-nitroarginine methyl ester led to a substantial decrease in acetylcholine output, the latter compound being more effective. The effect of L-NG-nitroarginine was abolished by simultaneous superfusion with L arginine. The effects of NO donors and of LY83583 suggest that NO increases acetylcholine release, probably by a cGMP-dependent mechanism. The effectiveness of nitric oxide synthase inhibitors shows that the activity of striatal neurons is under the permanent influence of nitric oxide, that leads, via a direct or indirect mechanism, to continuous enhancement of acetylcholine release. In conclusion, our findings suggest that NO synthesized in the ventral striatum acts as an intracellular messenger which modulates acetylcholine release. PMID- 7477428 TI - Further characterization of the putative 5-HT4 receptor mediating depolarization of the rat isolated vagus nerve. AB - A putative 5-HT4 receptor-mediated depolarization of the rat isolated vagus nerve has been studied using a grease-gap extracellular recording technique. Ondansetron (1 microM) was used to block the predominant 5-HT3 receptor mediated depolarization in this preparation and the effects of the 5-HT4 receptor antagonists DAU 6285 (endo-8-methyl-8-azabicyclo [3.2.1] oct-3-yl-2,3-dihydro-6 methoxy-2-oxo-1H-benzimidazole-1- carboxylate HCl); 0.3, 1.0 or 3.0 microM and SDZ 205-557 (2-methoxy-4-amino-5-chloro-benzoic acid 2-(diethylamino)-ethyl ester HCl); 0.1, 0.3 or 1.0 microM were studied on the residual, ondansetron-resistant, component of the response. The effects of the phosphodiesterase inhibitor isobutylmethylxanthine (IBMX) and of forskolin on the ondansetron-resistant response were also studied. Both DAU 6285 and SDZ 205-557 acted as competitive antagonists of the ondansetron-resistant response to 5-HT with pA2 values of 6.8 (6.7-7.1, n = 12) and 7.1 (6.9-7.5, n = 12) respectively. The vagus nerve was depolarized by IBMX (100 microM) or forskolin (10 microM), the effects being similar to the maximum response to 5-HT. In the presence of IBMX (100 microM) or forskolin (10 microM) the ondansetron-resistant component of the response to 5-HT was enhanced and the 5-HT3 receptor-mediated component reduced. These results with DAU 6285 and SDZ 205-557 are consistent with a 5-HT4 receptor-mediated mechanism of the ondansetron-resistant depolarizing response to 5-HT. PMID- 7477429 TI - Induction of giant miniature end-plate potentials during blockade of neuromuscular transmission by textilotoxin. AB - The present study investigated the action of textilotoxin, isolated from the venom of the Australian common brown snake Pseudonaja textilis, on neuromuscular transmission in isolated toad nerve-muscle preparations. Initial muscle twitch tension experiments revealed a triphasic pattern of changes in muscle tension and a irreversible binding action of textilotoxin (10 micrograms/ml) similar to other snake beta-neurotoxins. This was characterised by an initial depression of twitch tension, followed by a period of enhanced tension, eventually leading to a reduction in tension to complete neuromuscular blockade. These actions on muscle tension were investigated further by assessing the action of textilotoxin on end plate potential amplitude (EPP). This revealed a similar triphasic alteration of the nerve-evoked release of acetylcholine from the motor nerve terminal. These actions on acetylcholine release were confirmed to be of a presynaptic origin since the modal amplitude of miniature end-plate potentials (MEPPs) was not reduced and in twitch tension experiments the muscle still contracted in response to direct muscle stimulation when nerve-evoked release was completely blocked. Interestingly dramatic effects were observed on the spontaneous release of acetylcholine, including an marked increase in MEPP frequency, a skewing of the MEPP amplitude frequency histogram to the right, and a resultant increase in the number of 'giant' MEPPs. These results indicate that textilotoxin causes a presynaptic blockade of neuromuscular transmission involving a disruption of the regulatory mechanism that controls acetylcholine release. PMID- 7477430 TI - Characteristics and possible mechanisms of low-Na+ induced contractions in rat aorta. AB - The influence of reducing external Na+ concentration ([Na+]ex) upon vascular smooth muscle contractility was investigated using the rat isolated aorta. NaCl from the physiological saline solution (PSS) was replaced with either choline-Cl, sucrose, or LiCl to give the following [Na+]ex (mM): 115, 85, 55, and 25 (115NaPSS to 25NaPSS). Small reductions in [Na+]ex (115NaPSS) induced a biphasic contraction, comparable in amplitude with the control one induced by phenylephrine 10(-6) M. Elimination of the endogenous catecholamine participation using either phentolamine 10(-5) M or guanethidine 3.10(-6) M similarly reduces these contractions to 25% (sucrose replacement). A similar relaxing effect was obtained with D600 10(-5) M, an antagonist of the voltage operated Ca2+ channels (25-30% residual tension for all the substitutes). Large reductions in [Na+]ex (25NaPSS) induced contractions comparable in amplitude and shape, but less sensitive to phentolamine and guanethidine (residual tension 65-75%, sucrose replacement) and insensitive to D600 (all the substitutes). The Na+/K+ ATPase inhibitor ouabain (10(-4) M) elicited slowly developing contractions, the amplitude being 115% of the phenylephrine 10(-6) M control. Phenylephrine further contracted the 115NaPSS precontracted preparations, but was significantly less effective in 25NaPSS, although the precontraction levels were similar for the same substitute used. The amplitude of the superimposed phenylephrine contractions exhibited [Na+]ex dependence. Phenylephrine 10(-6) M failed to further contract the ouabain 10(-4) M precontracted rings. We conclude that relatively small reductions in [Na+]ex are able to induce contractions of rat aorta primarily through release of endogenous catecholamines, probably through neural Na+/Ca2+ exchange. Larger reductions in [Na+]ex appear to cause contraction through muscular Na+/Ca2+ exchange. PMID- 7477431 TI - Electrocardiological profile and proarrhythmic effects of quinidine, verapamil and their combination: a mapping study. AB - Quinidine and verapamil are widely used as antiarrhythmic agents and their combination is often used in the treatment of supraventricular tachycardia. This study was undertaken to clarify, whether these drugs exert proarrhythmic effects on the ventricles in therapeutic concentrations and whether possible arrhythmogenic effects might be enhanced by combination. Isolated rabbit hearts perfused according to the Langendorff technique were treated with increasing concentrations of quinidine (0.05 to 3.5 microM) or verapamil (5 to 50 nM) or of their combination (70:1 or 10:1, quinidine:verapamil) corresponding to common low, medium and high free therapeutic concentrations. The epicardial activation process was measured using a computer assisted mapping system for unipolar multichannel recording (256 channels simultaneously). Both substances prolonged the atrioventricular conduction time PQ. This effect was even more pronounced if the 70:1 combination was administered. The activation pattern was altered by both drugs and their combination to the same extent as became obvious from analysis of local activation vectors and of localisation of breakthroughpoints of epicardial activation for heart beats under control conditions and under drug treatment. The epicardial potential durations were prolonged by quinidine and to the same degree by the combinations, but not by verapamil alone. The total activation time was prolonged under the influence of quinidine and if the 70:1 combination was given. Both substances exerted a negative inotropic effect which was enhanced in an additive manner if both drugs were combined. In parallel the coronary flow was diminished.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7477432 TI - Alpha 2-adrenoceptor mediated inhibition of forskolin-stimulated cyclic AMP accumulation in isolated porcine palmar lateral veins. AB - The aim of this study was to use a 3H-adenine pre-labelling technique to characterise the effect of alpha 2-adrenoceptor activation on forskolin stimulated cyclic AMP accumulation in the isolated porcine palmar lateral vein. Forskolin (10(-7)-10(-4) M) stimulated 3H-cyclic AMP accumulation in the isolated porcine palmar lateral vein in a biphasic and concentration-dependent manner. In the absence of the cyclic AMP-selective phosphodiesterase inhibitor rolipram, forskolin stimulated 3H-cyclic AMP accumulation approximately 7-8 fold. The response reached a peak after 5 min. In the presence of rolipram (10(-5) M), basal 3H-cyclic AMP levels were approximately 70% higher than in its absence (basal: 1823 +/- 57 dpm; rolipram: 3088 +/- 229, n = 3) and forskolin (3 x 10(-5) M) stimulated 3H-cyclic AMP accumulation approximately 8 fold. The latter response reached a plateau 10 min after the addition of forskolin. In all subsequent studies, the tissues were incubated with forskolin (3 x 10(-5) M) for 5 min in the absence of rolipram. Noradrenaline (NA; 10(-9)-10(-4) M) and UK14304 (10(-9)-10(-4) M) inhibited forskolin-stimulated 3H-cyclic AMP accumulation in a concentration-dependent manner with mean pIC50 values of 7.61 +/- 0.37 (n = 4) and 7.76 +/- 0.23 (n = 5), respectively. With either NA or UK14304, the maximal inhibition of the forskolin response obtained was approximately 75%. Neither NA (10(-4) M) nor UK14304 (10(-4) M) altered basal 3H-cyclic AMP levels.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7477433 TI - The stimulatory effect of clonidine through imidazoline receptors on locus coeruleus noradrenergic neurones is mediated by excitatory amino acids and modulated by serotonin. AB - Clonidine and other imidazoline/oxazoline drugs, such as cirazoline and rilmenidine, have been shown to stimulate the activity of noradrenergic neurones in the locus coeruleus (NA-LC) by an alpha 2-adrenoceptor-independent mechanism through the activation of I-imidazoline receptors. The endogenous modulation of the stimulatory effect of clonidine on NA-LC neurones was further investigated after inactivation of alpha 2-adrenoceptors with N-ethoxycarbonyl-2-ethoxy-1,2 dihydroquinoline (EEDQ). In EEDQ-pretreated rats (6 mg/kg, i.p., 6 h), clonidine caused a rapid and dose-dependent (320-5120 micrograms/kg, i.v.) increase in the firing rate of NA-LC neurones (ED50 = 809 micrograms/kg, Emax = 90%). The stimulatory effect of clonidine on NA-LC neurones was completely blocked by pretreatment of rats with the excitatory amino acid receptor antagonist kynurenic acid (1-3 mumol in 10-30 microliters, i.c.v., 2-5 min before clonidine). In contrast, the stimulatory effect of clonidine on NA-LC neurones was potentiated by pretreatment with reserpine (5 mg/kg, s.c., 18 h) (Emax increased by 63%). Pretreatment with alpha-methyl-p-tyrosine (250 mg/kg, i.p., 24 h) did not alter the stimulatory effect of clonidine, but pretreatment with p-chloro-phenylalanine (400 mg/kg, i.p., 24 h) markedly enhanced the stimulatory effect of clonidine on NA-LC neurones (Emax increased by 139%). The present results indicate that the imidazoline receptor-mediated stimulatory effect of clonidine on NA-LC neurones is an indirect effect dependent on an excitatory amino acid pathway and modulated by an inhibitory serotonin mechanism. PMID- 7477434 TI - 5-HT1-like and 5-HT2A receptors mediate 5-hydroxytryptamine-induced contraction of rabbit isolated mesenteric artery. AB - The contractions induced by 5-hydroxytryptamine (5-HT) and the 5-HT1-like receptor agonist, sumatriptan, were investigated in the open ring preparations of rabbit mesenteric artery in order to characterize the 5-HT receptors. 5-HT induced concentration-dependent contractions. Sumatriptan did not induce any contraction of unstimulated rings, whereas it elicited concentration-dependent contractions in preparations given a moderate tone by a threshold concentration of prostaglandin F2 alpha (PGF2 alpha). Pargyline, cocaine or normetanephrine were without significant effect on the contractions induced by 5-HT and sumatripan. The 5-HT concentration-effect curve was clearly biphasic. Methiothepin (0.01 microM) shifted the both phases of the concentration-effect curve to the right. Ketanserin (0.1 microM) shifted the second, low affinity, phase and prazosin did not alter concentration-effect curve to 5-HT. The sumatriptan concentration-effect curve was shifted by methiothepin (0.01 microM) to the right (pKB = 9.19) but not by ketanserin (1 microM). Concentration-effect curves to 5-HT and sumatriptan were not affected by the 5-HT3 receptor antagonist tropisetron (1 microM). These results suggest that 5-HT1-like type receptors are responsible for the first phase of 5-HT-induced contraction and 5-HT2A receptor for the second phase, in rabbit mesenteric artery. Sumatriptan-induced contractions appear to be mediated by 5-HT1-like type receptors in this artery. These results also suggest that this kind of amplification may be a common feature of vascular 5-HT1-like type receptor as has been shown in other vascular segments such as rabbit femoral, iliac and renal arteries, and guinea-pig iliac artery. PMID- 7477435 TI - 5-HT4 receptor antagonist affinities of SB207710, SB205008, and SB203186 in the human colon, rat oesophagus, and guinea-pig ileum peristaltic reflex. AB - Functional 5-HT4 receptors have been reported to be present in numerous isolated tissue preparations including the rat oesophagus, guinea-pig ileum, and human colon. The pharmacological properties of the novel, potent and selective 5-HT4 receptor antagonists SB203186 (1-piperidinyl)ethyl 1H-indole 3-carboxylate), SB205008 (1-butyl-1-methyl-4-piperidinylmethyl)-8-amino-7-chloro-1,4-ben zodioxan 5- carboxylate iodide), and SB207710 (1-butyl-4-piperidinylmethyl)-8-amino-7-iodo 1,4 benzo-dioxan-5-carboxylate) were studied in these tissues. The nature of antagonism of the 5-HT-induced effects was investigated on the above isolated tissue preparations. 5-HT produced its effect with the following EC50 values: 400 +/- 0.4 nM (rat oesophagus, n = 20), 154 +/- 14 nM (guinea-pig ileum, n = 9) and 144 +/- 0.1 nM (human colon, n = 9). SB207710 (0.03-1 nM), SB205008 (1.0-10 nM), and SB203186 (10-100 nM) antagonised the 5-HT4 receptor-mediated relaxations of the carbachol-contracted rat isolated oesophagus against 5-HT with pKB values of 10.9 +/- 0.1, 9.5 +/- 0.1, and 9.0 +/- 0.1 respectively without effecting the maximum response. On the guinea-pig ileum peristaltic reflex preparation, SB207710 (0.01-1 nM) did not modify the reflex but it behaved as an antagonist of the 5-HT-induced facilitation with a pA2 value of 9.9 +/- 0.2.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7477436 TI - Early desensitization of somato-dendritic 5-HT1A autoreceptors in rats treated with fluoxetine or paroxetine. AB - Electrophysiological and autoradiographic approaches were used to assess possible changes in 5-hydroxytryptamine (serotonin) 5-HT1A receptors in the rat dorsal raphe nucleus after a subchronic treatment with fluoxetine or paroxetine, two specific serotonin reuptake inhibitors with antidepressant properties. Fluoxetine or paroxetine were injected daily (5 mg/kg, i.p.) for various time periods up to 21 days. Electrophysiological recordings performed 24 h after the last injection showed that the potency of the 5-HT1A receptor agonist, 8-OH-DPAT, to depress the firing of serotoninergic neurons in the dorsal raphe nucleus within brain stem slices was significantly reduced as early as after a 3-day treatment with either drug. The proportion of recorded neurons showing desensitization of somatodendritic 5-HT1A autoreceptors increased along the treatment from approximately 40% on the 3rd day to 60-80% on the 21st day. At no time during the treatment, was the specific binding of [3H]8-OH-DPAT (agonist radioligand) or [3H]WAY-100 635 (antagonist radioligand) to 5-HT1A receptors modified in the dorsal raphe nucleus or in other brain areas, suggesting that neither the density nor the coupling of these receptors to G-proteins were probably altered in rats injected with fluoxetine or paroxetine for up to 21 days. These results show that adaptive desensitization of somatodendritic 5-HT1A autoreceptors within the dorsal raphe nucleus can already be detected after a 3-day treatment with selective serotonin reuptake inhibitors. Rather than the desensitization per se, it may be the progressive increase in the number of serotoninergic neurons with desensitized 5-HT1A autoreceptors which plays a critical role in the (slowly developing) antidepressant action of these drugs. PMID- 7477437 TI - Increasing effect of ethanol on 5-HT3 receptor-mediated 14C-guanidinium influx in N1E-115 neuroblastoma cells. AB - N1E-115 mouse neuroblastoma cells were used to study the influence of ethanol on the 5-HT- and veratridine-induced influx of 14C-guanidinium via the 5-HT3 receptor channel and the fast sodium channel, respectively. Ethanol (10-100 mM) concentration-dependently increased the 5-HT-induced 14C-guanidinium influx, leaving the basal and veratridine (100 microM)-induced influx unaffected. The increasing effect of ethanol (100 mM) was observed at all 5-HT concentrations investigated; accordingly, ethanol increased the maximum response to 5-HT. Whereas in the absence of ethanol the concentration-response curve for 5-HT was bell-shaped, this was no longer the case when ethanol (100 mM) was present in the incubation buffer; the descending branch of the concentration-response curve for 5-HT at concentrations above 300 microM was virtually no longer observed. When, in the presence of substance P (10 microM) the 5-HT-induced 14C-guanidinium influx was already enhanced, the ability of ethanol (100 mM) to increase the 5-HT induced influx was considerably diminished (by 72%). Preincubation of N1E-115 cells with 5-HT caused a decay of the subsequent 5-HT response ("desensitization") which was dependent on the duration of preincubation; ethanol (100 mM) did not affect the rate of this decay of the 5-HT response. The 5-HT (30 microM)-induced 14C-guanidinium influx was also increased by methanol (100 mM) and n-propanol (100 mM). The rank order of the increasing effect of the n alkanols (at 100 mM) was: methanol < ethanol < n-propanol; i.e. the degree of enhancement increased with the lipophilicity of the alcohols.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7477438 TI - 5-HT1A receptor antagonists increase the activity of serotonergic cells in the dorsal raphe nucleus in rats treated acutely or chronically with citalopram. AB - In this study we have examined the acute effects of systemic administration of the selective serotonin reuptake inhibitor (SSRI), citalopram, in combination with either of the two selective 5-HT1A receptor antagonists, [(S)-5-fluoro-8 hydroxy-2-(dipropylamino)-tetralin [(S)-UH-301] or (+)-N-tertbutyl 3-(4-(2 methoxyphenyl)piperazin-1-yl)-2-phenyl-propionamide dihydrochloride [(+) WAY100135], on the activity of single 5-HT neurons in the dorsal raphe nucleus (DRN) of anesthetized rats using extracellular recording techniques. Acute administration of citalopram (0.3 mg/kg i.v.) significantly decreased the firing rate of DRN-5-HT cells most likely as a result of indirect stimulation of inhibitory somatodendritic 5-HT1A autoreceptors located on 5-HT cells in the DRN. This effect of citalopram was completely reversed by (S)-UH-301 (0.5 mg/kg i.v.) and partly by (+)-WAY100135 (0.5 mg/kg i.v.). Furthermore, the inhibitory effect of citalopram on the activity of 5-HT neurons was significantly attenuated by pretreatment with (S)-UH-301 (0.25 mg/kg i.v.) or (+)-WAY100135 (0.25 mg/kg i.v.). We have also studied the effects of (S)-UH-301 (0.03-0.50 mg/kg i.v.) on the firing rate of single DRN-5-HT cells in rats chronically treated with citalopram (20 mg/kg/day i.p. x 14 days). Administration of (S)-UH-301 significantly and dose-dependently increased the activity of 5-HT cells in citalopram-treated rats, but did not affect these neurons in saline-treated (1 ml/kg/day i.p. x 14 days), control rats. Our results thus suggest that 5-HT1A receptor antagonists can augment both the acute and chronic effects of citalopram on central serotonergic neurotransmission.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7477439 TI - Modulation of agonist binding by guanine nucleotides in CHO cells expressing muscarinic m1-m5 receptors. AB - In membranes prepared from CHO-m2 cells, inhibition of [3H]-N-methylscopolamine ([3H]NMS) binding by several muscarinic agonists resulted in competition curves with Hill slopes significantly different from unity. Addition of 5' guanylylimidodiphosphate (Gpp(NH)p) led to an increase in the IC50 value of the agonists with significant steepening of the inhibition curves. The shift in potency induced by Gpp(NH)p differed among the agonists with a rank order of oxotremorine-M = carbachol > oxotremorine > McN-A-343 = pilocarpine. In CHO-m4 membranes, Gpp(NH)p was less efficacious than in CHO-m2 membranes whereas no effect of the guanine nucleotide was found in membranes prepared from CHO-m1, m3, and -m5 cells. No major differences in the effect of Gpp(NH)p among agonists were found in CHO-m4 cells. Atropine binding was not affected by the guanine nucleotide. Together, these results indicate that coupling of G-proteins to muscarinic receptors linked to inhibition of cyclic adenosine monophosphate (cAMP) (m2 and m4) but not of those linked to phosphoinositol turnover (m1, m3 and m5) can be perturbed by Gpp(NH)p. The differential effects observed with Gpp(NH)p between agonist binding to m2 and m4 receptors appear to be receptor specific and may reflect differences in the G proteins activated by these receptors in CHO cells. PMID- 7477440 TI - Conditional involvement of muscarinic M1 receptors in vagally mediated contraction of guinea-pig bronchi. AB - The involvement of ganglionic muscarinic M1 receptors in vagally induced bronchoconstriction in guinea-pig airways is controversial. Therefore, we studied the effects of the M1-selective muscarinic receptor antagonist pirenzepine on vagus nerve (VNS, preganglionic) and electrical field stimulation (EFS, postganglionic)-induced contractions of the guinea-pig main bronchus under various experimental conditions. Using identical stimulation parameters for VNS and EFS (8V, 30 Hz, 0.5 ms, 5s every min), the amplitude of the VNS-induced twitch contractions was 30.4% of the EFS-induced responses, and pirenzepine showed 2.3-fold selectivity (pIC50-values 6.45 and 6.09, respectively) to inhibit vagally induced contractions. With the stimulation frequency for EFS lowered to match contraction levels obtained using VNS, pirenzepine was equipotent to inhibit both types of response at M3 receptor-selective concentrations, suggesting that M1 receptors are not involved. By contrast, when the stimulation episode was prolonged until plateau contraction (10-20 s), in the presence of the nicotinic antagonist hexamethonium (5 microM), the M2 receptor antagonist AQ-RA 741 (0.1 microM) and the beta-adrenoceptor antagonist timolol (1 microM), and again using matched VNS- and EFS-induced contraction levels, pirenzepine inhibited nerve stimulation-evoked responses in a biphasic manner, yielding pIC50 values of 8.12 (indicative of M1 receptor blockade) and 6.43 (indicative of M3 receptor blockade) for the first and second phase, respectively, while postganglionic stimulation showed a purely monophasic inhibition (pIC50 = 6.32). These results show that facilitatory muscarinic M1 receptors are involved in vagally mediated contraction of guinea-pig bronchi, under conditions of elevated neurotransmission and partial nicotinic receptor blockade. PMID- 7477441 TI - Pharmacological characterization of muscarinic acetylcholine binding sites in human and bovine cerebral microvessels. AB - The aim of this study was to investigate the presence of muscarinic receptors in human brain microvessels (MVs) and capillaries (CAPs) and, further, to pharmacologically characterize these receptors as well as those in bovine cerebral microvascular beds. For this purpose, the binding of [3H]N-methyl scopolamine ([3H]NMS) was assessed in isolated human and bovine cerebral MVs and CAPs and competition studies were performed against [3H]NMS binding with several well characterized muscarinic antagonists. The antagonist cerebrovascular affinity constants (pKD) were determined with the computer-fitting software LIGAND and then compared by correlation analyses to their reported affinities (pKi) at the five cloned muscarinic receptors. The specific binding of [3H]NMS to human and bovine MVs and CAPs was saturable, of high affinity and competitively inhibited by muscarinic antagonists. Heterogeneous populations of muscarinic binding sites were found in the microvascular tissues from both species. In human cortical MVs, the pharmacological binding profile obtained from various muscarinic receptor antagonists was best correlated to that of the cloned ml (r = 0.95; p < 0.001) and less so m5 (r = 0.77; p = 0.025) receptor subtypes while in bovine MVs, the presence of the m1 subtype was strongly suggested. Cerebrovascular affinities obtained for selected muscarinic antagonists in single preparations of human and bovine CAPs were suggestive of the presence of M1/m1 and M3/m3 receptor subtypes, and possibly the m5 subtype in bovine CAPs. The detection of M1/m1, M3/m3 and possibly m5 muscarinic receptor subtypes in brain microcirculation is consistent with reports where these receptors have been shown to mediate vasoconstriction, vasodilatation, and activation of nitric oxide synthase, respectively.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7477443 TI - Potentiation by enalaprilat of fenoldopam-evoked natriuresis is due to blockade of intrarenal production of angiotensin-II in rats. AB - We have previously shown that the natriuretic response to DA-1 receptor agonist fenoldopam is markedly potentiated by angiotensin converting enzyme inhibitor captopril. Since inhibition of angiotensin converting enzyme can lead to decreased production of angiotensin-II and increased levels of kinins (e.g., bradykinin), it is likely that both of these mechanisms might be involved in this phenomenon. However, it is not known whether and to what degree the accumulation of kinins contributes to the overall potentiation of natriuretic response to fenoldopam seen during angiotensin converting enzyme inhibition. In the present study, we have examined the effect of angiotensin converting enzyme inhibitor enalaprilat and angiotensin-II receptor antagonist losartan as well as bradykinin 2 receptor antagonist HOE 140 on fenoldopam-induced natriuresis. Intravenous infusion of fenoldopam (1 microgram/kg/min) for 30 min produced significant increases in urine output and urinary sodium excretion without causing any changes in glomerular filtration rate, renal blood flow and mean arterial blood pressure, a phenomenon suggestive of a direct tubular site of action. In animals treated with either the angiotensin converting enzyme inhibitor enalaprilat or angiotensin-II receptor antagonist losartan, the diuretic and natriuretic effects of fenoldopam were potentiated to a similar degree. Whereas no significant changes in glomerular filtration rate occurred when fenoldopam alone was given to control rats, in animals treated with either enalaprilat or losartan, fenoldopam produced a modest but significant increase in glomerular filtration rate. In a separate group of animals, the effects of bradykinin-2 receptor antagonist HOE 140 on potentiation of fenoldopam-induced natriuresis by enalaprilat was examined.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7477444 TI - The non-peptide NK1 receptor antagonist SR140333 produces long-lasting inhibition of neurogenic inflammation, but does not influence acute chemo- or thermonociception in rats. AB - In anaesthetized rats, the neurokinin (NK)1 receptor antagonist SR140333 (10-1000 micrograms/kg) stereo-selectively inhibited mustard oil-induced plasma protein extravasation in the dorsal skin of the hind paw. After s.c. administration of SR140333, inhibition of plasma protein extravasation was maximal 3 h after injection. A dose of 0.1 mg/kg i.v. or 1.0 mg/kg s.c. produced long-lasting inhibition which was still significant 24 h after treatment. Since systemic administration of SR140333 has been shown to inhibit nociceptive responses in anaesthetized rats, we wanted to evaluate a possible effect of SR140333 on chemo- and thermonociception in conscious rats. SR140333 (100 micrograms/kg s.c.) did not reduce the behavioral response of rats to the irritant effect of capsaicin in the wiping test, nor did it affect the thermal nociceptive threshold in the plantar test. Furthermore, the decrease in thermal nociceptive threshold which was produced by intraplanter injection of PGE2, and which has been shown to be entirely dependent on capsaicin-sensitive afferents, was not affected by treatment with this NK1 receptor antagonist. These results show that systemic administration of SR140333, at doses which cause inhibition of neurogenic inflammation, has no detectable effect on acute chemo- or thermonociception in conscious rats. PMID- 7477442 TI - The striatal dopaminergic catalepsy mechanism is not necessary for the expression of pontine catalepsy produced by carbachol injections into the pontine reticular formation. AB - We have found previously that microinjections of carbachol into the pontine reticular formation (PRF) of rats induce an intense cataleptic state which is similar behaviorally with the catalepsy induced by systemic administration of neuroleptic drugs. In the experiments described in the present article we studied the possibility that the pontine carbachol catalepsy is generated via the intermediary of the dopaminergic cataleptogenic mechanism in the striatum. To this purpose we produced kainic acid lesions in the striatum and in the output stations of the striatal cataleptogenic mechanism-substantia nigra reticulata and the VM thalamic nucleus. Catalepsy was tested after systemic haloperidol (2 mg/kg) and pontine microinjections of carbachol (5 micrograms/1 microliter) before and after the kainic lesions. The cataleptogenic effect of carbachol injected in the pons was not attenuated by any of the three types of lesions. On the contrary, the cataleptogenic effect of haloperidol was greatly attenuated by the same lesions. These results suggest that the pontine catalepsy produced by microinjections of carbachol in PRF is generated independently of the dopaminergic cataleptogenic mechanism in basal ganglia. PMID- 7477445 TI - Enhancement of adenosine A1 receptor functions by benzoylthiophenes in guinea pig tissues in vitro. AB - Previous reports on a series of benzoylthiophenes, including PD 81,723 [2-amino 4,5-dimethyl-3-(3-trifluoromethyl-benzoyl) thiophene], have shown specific enhancement of agonist binding at the adenosine A1 receptor. We have studied the effects of two substituted benzoylthiophenes, PD 78,416 (thieno[2,3-c]pyridine 6(5H)-carboxylic acid, 2-amino-3-benzoyl-4,7-dihydro-ethyl ester) and RS-74513 000 [2-amino-4-ethyl-5-methyl-3-(3-trifluoro-methyl-benzoyl) thiophene] on response elicited by adenosine A1 receptors in isolated guinea pig left atrium and ileum. In the electrically paced left atrium, PD 78,416 antagonized negative inotropic effect elicited by the agonist CPA [N6-cyclopentyladenosine] with a pKB value of 6.2 +/- 0.2 (n = 4). At a low concentration which had no antagonistic effect (0.1 microM), PD 78,416 enhanced the effect of CPA. The concentration response curve to CPA was shifted leftward by 5.1 fold (95% confidence limits 2.4 11.2). In field stimulated isolated ileum, PD 78,416 (0.1, 0.3, 1 microM) did not enhance or antagonize effects of CPA. At concentrations above 1 microM, PD 78,416 decreased electrically induced contraction. This effect was not sensitive to adenosine deaminase and was not antagonized by the A1 antagonist CPX [8 cyclopentyl-1,3-dipropyl-xanthine] (1 microM). Unlike PD 78,416, RS-74513-000 (0.01, 0.1, 1, 3, 10 microM) did not antagonize or enhance effects of CPA in the left atrium. However, effects of CPA in ileum were enhanced by RS-74513-000 (1 and 3 microM).(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7477446 TI - Evidence that BKCa channel activation contributes to K+ channel opener induced relaxation of the porcine coronary artery. AB - The rank order of potency of a series of benzopyran and cyanoguanidine K+ channel openers (KCOs) for causing relaxation of the PGF2 alpha-precontracted porcine coronary artery was determined. Glyburide, an inhibitor of KATP channels, showed an apparent competitive inhibition of the vasorelaxant activity of the KCOs. The pA2 values of glyburide when cromakalim and CGP 14877 (P1060) were used as vasorelaxants were 7.66 and 7.83, respectively. Charybdotoxin (40 nM), an inhibitor of BKCa channels, also caused a significant inhibition of the cromakalim mediated relaxation of the porcine coronary artery. In order to clarify the site of action of these KCOs, we identified a K+ channel current in single porcine coronary arterial cells and measured channel activity in the presence of these compounds. The prominent K+ ion current in these cells had characteristics typical of the conventional large Ca(2+)-activated K+ channel (BKCa) present in other smooth muscle cells. Using symmetrical K+ concentrations, the channel had a conductance of 214 pS and was found to be sensitive to [Ca2+]i and membrane potential. The KCOs were found to reversibly increase the open probability (P(o)) of the channel without changing channel conductance. The potency of the KCOs to increase K+ channel opening was similar to the potency of these compounds to cause coronary artery relaxation. These results indicate that the porcine coronary artery contains the BKCa channel and that this channel, along with other types of K+ channels (KATP), mediate the vasorelaxant effects of K+ channel openers. PMID- 7477447 TI - Effects of cromakalim or glibenclamide on arrhythmias and dispersion of refractoriness in chronically infarcted in anesthetized dogs. AB - The proarrhythmic effects of the ATP-sensitive potassium channel modulators cromakalim (n = 10; 0.01 to 0.3 mg/kg i.v.), glibenclamide (n = 10; 0.3 to 10 mg/kg i.v.) or volume equivalents of vehicle (n = 10) were evaluated in post infarcted anaesthetised dogs. Dogs were anaesthetised, subjected to an anterior apical myocardial infarction, and allowed to recover. At 7.4 +/- 0.7 days post infarction, animals were anaesthetised again, electrophysiologic measurements (effective refractory periods, QT-intervals and ventricular fibrillation thresholds) were taken, and animals were tested for arrhythmias using a programmed electrical stimulation protocol. Only animals that did not have programmed electrical stimulation-inducible arrhythmias were used. Ventricular fibrillation thresholds were determined twice, once before the first dose then after the last dose of drug. At the end of the experiment, animals were subjected to ligation of the left circumflex coronary artery and survival was measured over the next two hours. Cromakalim significantly increased heart rate and decreased blood pressure. Although cromakalim significantly reduced effective refractory periods, it neither increased electrical dispersion, as determined by the standard deviation or coefficient of variance of the effective refractory period, nor did it enhance inducibility (0 out of 10 in both vehicle and cromakalim treated animals), change ventricular fibrillation thresholds, or reduce sudden death survival relative to vehicle. Glibenclamide did not increase electrical dispersion, but slightly increased the incidence of programmed electrical stimulation-induced arrhythmias (3 out of 10), and lowered ventricular fibrillation thresholds values. However, these changes were not statistically significant. Glibenclamide did not significantly affect survival relative to vehicle.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7477448 TI - Comparison of corelease of noradrenaline and ATP evoked by hypogastric nerve stimulation and field stimulation in guinea-pig vas deferens. AB - Contractions and overflow of tritium and ATP elicited by hypogastric nerve stimulation (HNS) and field stimulation (FS) were studied in the guinea-pig isolated vas deferens preincubated with [3H]-noradrenaline. ATP was measured by means of the luciferin-luciferase technique. HNS and FS elicited contraction, tritium overflow and ATP overflow. HNS at supramaximal current strength produced smaller responses than did FS at supramaximal current strength (210 pulses/7 Hz). Supramaximal HNS and submaximal FS were used in the remainder of the study. Prazosin (0.3 mumol/l) reduced contractions and the overflow of ATP elicited by both HNS and FS; the evoked overflow of tritium was not changed (210 pulses/7 Hz). Combined administration of prazosin (0.3 mumol/l) and suramin (300 mumol/l) abolished contractions and reduced the overflow of ATP elicited by both HNS and FS slightly more than did prazosin alone; tritium overflow again was not changed (210 pulses/7 Hz). Contractions, tritium overflow and ATP overflow increased with the frequency of both HNS and FS (from 7 to 25 Hz; 210 pulses); the increase in ATP overflow with frequency was more marked than the increase in tritium overflow. The preferential increase of ATP overflow with the frequency of HNS and FS persisted in the combined presence of prazosin (0.3 mumol/l) and suramin (300 mumol/l). The study confirms for HNS, a more physiologic way of sympathetic nerve stimulation, several observations previously obtained with FS. First, HNS-evoked ATP release is detectable as an overflow of ATP into the superfusion fluid.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7477450 TI - Tularemia fatality. PMID- 7477449 TI - Sub-nanomolar concentrations of ciguatoxin-1 excite preganglionic terminals in guinea pig sympathetic ganglia. AB - The actions of low concentrations of ciguatoxin-1 (CTX-1, 0.2-0.8 nM) in guinea pig sympathetic ganglia have been analysed using intracellular recording techniques in vitro. The effects of CTX-1 were graded with concentration but sensitivity varied markedly between neurones in the same preparation. Other than an initial transient (approximately 10 min) depolarization of some ganglion cells accompanied by an increase in input resistance, passive electrical properties did not significantly differ from controls. Amplitude and threshold of action potentials evoked by depolarizing current and threshold, latency and form of the initial responses to nerve stimulation were also not affected. Exposure to CTX-1 generated marked increases in the frequency of spontaneous excitatory synaptic potentials which often occurred in bursts (15-66 Hz) of similar amplitudes. Single stimuli to incoming nerves produced repetitive synaptic responses arising from preganglionic, but not from peripheral afferent, axons. Following brief (< 5 min) exposure to CTX-1, these effects declined over 30 min but, after longer exposure (> 15 min), they persisted for several hours despite continuous washing. All activity generated by CTX-1 was significantly reduced or abolished by d tubocurarine (10(-5)-10(-4) M), hexamethonium (10(-5) M), tetrodotoxin (10(-7) 10(-6) M), omega-conotoxin (10(-7) M), reduced Ca2+ (0.1 mM)/raised Mg2+ (10 mM), raised Ca2+ (6 mM) or raised Mg2+ (25 mM). The data suggest that CTX-1 activates preganglionic axons by modifying the voltage sensitivity of a subpopulation of Na+ channels. Effects on these unmyelinated axons occur at much lower concentrations than have been reported to affect myelinated ones. Many of the symptoms of ciguatera poisoning might be explained by activity in autonomic and perhaps other unmyelinated nerve terminals. PMID- 7477451 TI - Race, education, and mortality in North Carolina. PMID- 7477452 TI - Diabetes in North Carolina. Directions for the future. PMID- 7477454 TI - Adult depression. A treatable condition. PMID- 7477453 TI - Use of subdermal contraceptive implants in a community-based family planning program. Experience after two years. PMID- 7477455 TI - The interdisciplinary geriatric team evaluation project. A new approach to the delivery of geriatric medicine in geographically remote locations. PMID- 7477456 TI - Adverse drug reactions in a state psychiatric hospital. PMID- 7477457 TI - Doctor as witness, doctor as juror. Going to court in a non-defendant, non adversarial role. PMID- 7477458 TI - It's the cortex, stupid. PMID- 7477459 TI - Will there be members of the North Carolina Medical Society in the future? PMID- 7477461 TI - The drug testing dilemma. PMID- 7477460 TI - Snapshot of an illness. PMID- 7477462 TI - Tularemia in North Carolina. PMID- 7477463 TI - Infant mortality under scrutiny. PMID- 7477464 TI - The burdens of abuse and illness. PMID- 7477466 TI - A community policy on futility? A conversation of the North Carolina community. PMID- 7477465 TI - When you don't know what to say, don't say anything. PMID- 7477467 TI - Beginning a conversation on medical futility in North Carolina. PMID- 7477468 TI - North Carolina hospitals' policies on medical futility. PMID- 7477469 TI - The ethical issues of futility from a community perspective. PMID- 7477470 TI - Lessons from communities in conversation. The Colorado experience. PMID- 7477471 TI - Real futility. Historical beginnings and continuing debate about futile treatment. PMID- 7477472 TI - Exercise and older adults. PMID- 7477473 TI - Ethical and legal barriers to futility policies. PMID- 7477474 TI - Futility policies and the law. PMID- 7477476 TI - Futility policies from the caregiver's perspective. PMID- 7477475 TI - The legislative role in defining medical futility. PMID- 7477477 TI - Ethical disputes over futile treatments. PMID- 7477478 TI - Religious aspects of futile treatment. PMID- 7477479 TI - Decisions about futile treatment in an intensive care nursery. PMID- 7477480 TI - Secondary payor rules, exploring managed care hot topics for NRAA. PMID- 7477481 TI - Dialysis in a time of war: a first-hand look. Part I. Interview by Cynthia K Lefton. PMID- 7477482 TI - A question of ethics? Purchasing a kidney transplant: a story of life regained. PMID- 7477483 TI - Organ shortage makes payment tempting. PMID- 7477484 TI - A question of ethics? International medical community condemns death row donations in China. PMID- 7477485 TI - Challenges facing dialysis patients requiring physical rehabilitation. PMID- 7477486 TI - Improve OPO service, improve donation. PMID- 7477487 TI - [WGBO (Law on Medical Treatment Agreement): starting and discontinuing hemodialysis in a 90-year-old patient]. PMID- 7477488 TI - [The standard 'Allergic and Hyperreactive rhinitis' of the Dutch Society of Family Physicians; reaction from the viewpoint of family practice]. PMID- 7477489 TI - [The standard 'Allergic and hyperreactive rhinitis' of the Dutch Society of Family Physicians; reaction from the allergological viewpoint]. PMID- 7477490 TI - [Synopsis of the standard 'Allergic and hyperreactive rhinitis' of the Dutch Society for Family Physicians]. PMID- 7477491 TI - [A stamp with emphasis points as practical aid in the information interview in the framework of the Law on Medical Treatment Agreement]. PMID- 7477492 TI - [Good surgery results in older patients with stage 1 lung carcinoma]. AB - OBJECTIVE: To determine the influence of age on survival of patients undergoing resection for lung cancer. DESIGN: Retrospective. SETTING: South-western part of the Netherlands. METHODS: Follow-up information was gathered on patients who had undergone resection for stage I non-small cell lung cancer from 1984 through 1990 and had been registered by the Rotterdam Cancer Registry. Operative mortality, 2 year and 5-year survival in the age groups 20-59, 60-69 and 70 years and older were compared. RESULTS: Of the 630 patients (median age of 66 years) one-third was 70 years or older. Operative mortality was mainly determined by the type of operation: 6% after pneumonectomy versus 1% after lesser resections. Five-year survival declined with age from 65% to 48% and 43% (p < 0.01). After adjustment for non-related causes of death this difference decreased: 68%, 55%, 61% (p = 0.15). The main prognostic factor was tumour size. CONCLUSION: Chronological age has a limited influence on the survival of patients with lung cancer and does not preclude surgical treatment. PMID- 7477493 TI - [Characteristics of patients with the diagnosis 'mental strain' in family practice]. AB - OBJECTIVE: To investigate the characteristics of general practitioners' patients given the diagnosis of 'surmenage' (French for 'nervous breakdown'). DESIGN: Descriptive. SETTING: General practices in Almere, the Netherlands. METHOD: Four groups of general practice attenders were selected based on the general practitioners' diagnosis: a group of surmenage patients (n = 106), a group of somatic patients without apparent psychological problems (n = 159), a group of patients with a psychosocial diagnosis (n = 136) and a group of patients with a psychiatric diagnosis (n = 57). Symptoms, social functioning, background data, life events and problems were collected by questionnaires. RESULTS: Most surmenage patients were employed young adults. They experienced more work or study related problems than the other patients. They also experienced more other problems than the somatic patients but not more than the patients with a psychosocial or psychiatric diagnosis. More than half of the surmenage patients were on sick leave. The most important symptoms in surmenage patients were nervousness, brooding, fatigue and insomnia. On the basis of their symptoms the surmenage patients could be distinguished well from the somatic and psychosocial patients, but not from the patients with a psychiatric diagnosis. The surmenage patients had a shorter duration of symptoms than the patients with a psychosocial or a psychiatric diagnosis. The group of surmenage patients was very heterogeneous. CONCLUSION: The diagnosis of surmenage is associated with nonspecific psychological symptoms, a relatively recent onset, being employed, experiencing problems related to work or study, and being on sick leave. Patients diagnosed as suffering from surmenage by their general practitioner, form a heterogeneous group. PMID- 7477494 TI - [The longitudinal development of smoking behavior in men and women between 13 and 27 years and the relationship with biological risk factors for cardiovascular diseases]. AB - OBJECTIVE: To describe the longitudinal development of smoking behaviour from adolescence (age 13 years) into young adulthood (age 27 years) and to analyse the longitudinal relationship between smoking behaviour and biological risk factors for cardiovascular disease. DESIGN: Longitudinal. SETTING: Amsterdam school children. METHODS: This study was carried out as part of the Amsterdam Growth and Health Study, an observational longitudinal study. Six measurements were carried out over a period of 15 years in 84 males and 98 females, and included a questionnaire regarding smoking behaviour and blood analysis for cholesterol parameters. Tracking analysis and generalized estimating equations analysis were used. RESULTS: Both the percentage of smokers and the amount of tobacco smoked by smokers increased until the age of 21 years. After that smoking behaviour remained more or less stable in males and decreased slightly in females (period effect). Between 16 and 21 years half the number of persons tried to stop smoking but failed to succeed. The development of smoking behaviour appeared to be related to low systolic blood pressure and to development of a negative risk profile regarding hypercholesterolaemia (i.e. low high-density lipoprotein (HDL) cholesterol and high ratio of total cholesterol and HDL cholesterol). There was no relationship between smoking and the amount of subcutaneous fat. PMID- 7477495 TI - [Cholecystitis in infants]. AB - In two infants presenting with non-specific abdominal symptoms, three months and four weeks of age, respectively, cholecystitis was suspected on ultrasound examination of the abdomen and confirmed and treated by surgery. In the first patient the cholecystitis was secondary to cholelithiasis. The second patient was shown to have an infected gallbladder due to an anatomical anomaly of the cystic duct. It is concluded that even in young infants, cholecystitis as a complication of congenital or neonatal pathology may underlie abdominal symptoms. Ultrasonography of the abdomen is an important diagnostic aid and cholecystectomy is the treatment of choice. PMID- 7477496 TI - [The Law on Medical Treatment Agreement: common ground with medical education and medical specialist]. PMID- 7477497 TI - [Preoperative internal medicine consultation. I. General aspects and management in diabetes mellitus and obstructive lung disease]. PMID- 7477498 TI - [Intra-anal application of isosorbide dinitrate in chronic anal fissure]. PMID- 7477499 TI - [Puerperal complication: 'Postpartum headache' (thrombosis of the superior sagittal sinus)]. PMID- 7477500 TI - [Puerperal complication: 'Postpartum headache' (thrombosis of the superior sagittal sinus]. PMID- 7477501 TI - [Consensus depression in adults]. PMID- 7477502 TI - [Femoral neck fractures in bicyclists due to clipless pedals]. PMID- 7477503 TI - [Deep neck abscess; a treacherous and life-threatening disorder]. PMID- 7477504 TI - [Nursing home medicine, a new medical specialism]. PMID- 7477505 TI - [Microalbuminuria in diabetes mellitus]. PMID- 7477506 TI - [Treatment of patients with trophoblastic tumors in the Academic Medical Center: 31 patients in 10 years, 1983-1992]. AB - OBJECTIVE: Evaluation of the treatment of patients with gestational trophoblastic disease. SETTING: Academic Medical Centre, Amsterdam. DESIGN: Descriptive. METHOD: Clinical data from all patients treated between 1983 and 1992 were collected. RESULTS: Thirty-one patients were treated in this period. Eighteen patients had persistent trophoblast after a molar pregnancy, 7 had choriocarcinoma after a normal pregnancy and in 6 patients the obstetrical history was unclear. The initial treatment strategy was chosen on the basis of prognostic factors; low-risk patients received mono-chemotherapy (Methotrexate) and high-risk patients were treated with polychemotherapy (EMA/CO). In 15 cases adjuvant therapy was necessary. Complete remission was achieved in 29 patients, 2 patients died. The adverse effects of the chemotherapy were generally mild. CONCLUSION: Persistent trophoblastic disease has an unpredictable presentation and course. Therefore treatment should preferably be given in a reference centre. PMID- 7477507 TI - [From urostoma to bladder replacement: current possibilities in urinary diversion]. AB - OBJECTIVE: To analyse 48 urinary diversions in an oncological patient population. DESIGN: Retrospective study. SETTING: The Dutch cancer institute 'Nederlands Kanker Instituut/Antoni van Leeuwenhoek Ziekenhuis' in Amsterdam, the Netherlands. METHOD: The medical records of all patients who had a urinary diversion in the period 1989-1993 were analysed for type of diversion, indication for this specific type and postoperative complications. RESULTS: In 48 patients (24 women and 24 men with a mean age of 62.1 years) a urinary diversion was constructed because of cystectomy for bladder cancer (n = 27), cystectomy in total pelvic exenteration (13), total urinary incontinence following radiotherapy (5), urethral cancer (1) or local palliation (2). Twenty-one diversions according to Bricker, 25 continent reservoirs according to Rowland and (or) neo-bladders, I ureterosigmoidostomy and I continent access to the bladder (Mitrofanoff procedure) were done. The standard diversion according to Bricker was preferred in the following circumstances: extensive intestinal resections (also previous ones), older age, fear of incontinence or no need for continent diversion. Specific problems of the new diversion techniques were difficulties of patients with catheterising the reservoir and folic acid deficiency. All other postoperative problems were related to the extensiveness of the primary oncological procedure rather than to the diversion technique. CONCLUSION: Apart from a few contraindications such as shortage of intestine, tumour growth into the urethra and patients' inability or unwillingness to catheterize themselves, there are no objections to the new diversion techniques (continent reservoir/neo bladder). Patients being considered for urinary diversion ought to be informed about these new techniques. PMID- 7477508 TI - [Serum screening for fetal Down syndrome in pregnant women with an indication for prenatal diagnosis because of age: fewer amniocenteses]. AB - OBJECTIVE: To examine the influence of maternal serum screening for foetal Down's syndrome (DS) on the number of amniocenteses in women with an indication for prenatal diagnosis because of age > or = 36 years. DESIGN: Longitudinal descriptive study. SETTING: Department of Obstetrics and Gynaecology, University Hospital Groningen. METHOD: Between October 1, 1990 and March 31, 1994, sera from 693 women, 36 years or older with a singleton pregnancy, were tested (alpha foetoprotein and human chorionic gonadotrophin) to calculate the likelihood of their having a foetus with DS. RESULTS: 195 pregnant women (28%) were screen positive (risk of having a foetus with DS > or = 1:250); 105 of these (54%) chose to have an amniocentesis. Of the remaining 498 (screen-negative) women, 22 (4%) chose to have an amniocentesis. All 7 cases of DS were in the screen-positive group. CONCLUSION: Maternal serum screening in women aged > or = 36 years can markedly reduce the number of invasive prenatal diagnostic procedures, with a minimal reduction in the detection of DS foetuses. It is advisable to offer this form of screening to all women in this age group. PMID- 7477509 TI - [Chondrosarcoma of the bony thoracic wall]. AB - An 85-year-old man was admitted because of a slowly growing mass in his left flank, which had been present for 16 years. A very large chondrosarcoma was diagnosed, weighing 8.5 kg. After radical resection of the tumour the patient recovered well. No recurrences developed during follow-up. PMID- 7477510 TI - [Current guidelines for the prevention and treatment of diabetic nephropathy. International Diabetes Federation]. PMID- 7477511 TI - [Replacement of preservation agent Kathon CG in cosmetics by Euxyl K 400; indeed out of the frying pan into the fire]. PMID- 7477512 TI - [Scientific research in nursing home medicine: a prerequisite for professionalization]. PMID- 7477513 TI - [What is 'Doctors without Borders' doing in Bosnia?]. PMID- 7477514 TI - [3 patients with autoimmune hemolytic anemia in a peripheral hospital]. PMID- 7477516 TI - [Urinary incontinence: a gynecological and a urological problem]. PMID- 7477515 TI - [Selective targetting of antiviral agents to the liver]. PMID- 7477517 TI - [Erection disorders and priapism due to drugs]. PMID- 7477518 TI - [Guidelines for the diagnosis and treatment of erectile disorders. NVIO (Dutch Society for Impotence Research) and NVU (Dutch Society for Urology)]. PMID- 7477519 TI - [Heart surgery and surgical mortality in 1982 and 1992 in the St. Antonius Ziekenhuis, Nieuwegein. Are indication criteria changing?]. AB - OBJECTIVE: To determine the changes in the open heart surgical procedures and hospital mortality between 1982 and 1992. DESIGN: Retrospective investigation. SETTING: St. Antonius Hospital, Nieuwegein. METHOD: A comparison of the open heart surgical procedures, the hospital mortality and the age distribution of the operated patients was made by means of the database of the department of Cardiopulmonary Surgery. RESULTS: The total number of open heart procedures showed a small increase; however, there was a considerable increase in the number of combined procedures of coronary revascularisations with valve surgery, coronary revascularisation reoperations, closure of ventricular septal ruptures, rhythm procedures and reconstruction of the thoracic aorta. Although the mean age of the patients increased by about 6 years, hospital mortality was much lower in 1992. CONCLUSIONS: Between 1982 and 1992 mean patient age increased considerably, the open heart procedure became more extensive and the hospital mortality decreased. PMID- 7477520 TI - [Diagnosis and therapy of urinary incontinence in Dutch gynecologic practice; a questionnaire study]. AB - OBJECTIVE: Assessment of modalities in diagnosis and treatment of female urinary incontinence by Dutch gynaecologists. DESIGN: Written questionnaire. SETTING: All officially registered Dutch gynaecological practices. METHOD: All 138 gynaecological practices were sent a multiple choice questionnaire. One representative of the practice was requested to respond. Differences were evaluated using the chi 2 test. RESULTS: The response was 87% (93% of the teaching hospitals and 86% of the non-teaching hospitals). Diagnostic and treatment modalities of female urinary incontinence by Dutch gynaecologists proved to vary. Urodynamics are primarily under control of urologists. The Burch colposuspension as operative treatment of stress incontinence is fairly popular among Dutch gynaecologists, but the less successful anterior colporrhaphy with Kelly plication is also still performed rather frequently. Co-operation between gynaecologist and urologist is mainly practised in teaching hospitals at meetings at which patient histories and urodynamics are discussed. CONCLUSION: There is need for systematism in diagnosis and treatment of female urinary incontinence and for intensification of co-operation between gynaecologists and urologists. Adequate training and postgraduate teaching courses in diagnosis and treatment of female urinary incontinence should be provided for both specialisms. PMID- 7477521 TI - [Intestinal pseudo-obstruction during use of baclofen]. AB - In a 75-year-old man acute intestinal pseudo-obstruction was observed during treatment with baclofen 20 mg daily. After discontinuation of the baclofen he recovered completely. No other cause of intestinal obstruction could be demonstrated. Baclofen is an agonist of spinal GABA receptors. It is used in the treatment of spasticity caused by multiple sclerosis or other diseases of the spinal cord, in particular traumatic lesions. Physicians should be aware of this possible adverse effect of baclofen. PMID- 7477522 TI - [Assessment of reporting procedure euthanasia]. PMID- 7477523 TI - [A fair trial? The European randomized prostate cancer study]. PMID- 7477524 TI - [Diagnosis of lymph node metastasis of head-neck carcinomas]. PMID- 7477525 TI - [Biliary tract carcinoma: operate or irradiate?]. PMID- 7477526 TI - [Repressed memories]. PMID- 7477527 TI - [Thrombosis of the cerebral veins and sinuses in 62 patients]. PMID- 7477528 TI - [An unusual complication of pneumococcal pneumonia: tamponade due to purulent pericarditis]. PMID- 7477529 TI - [Quality of medical care for persons with a mental handicap]. PMID- 7477530 TI - [Primary liver tumors in children]. PMID- 7477531 TI - [Thrombopoietin produced molecularly-biologically]. PMID- 7477532 TI - [Malaria in a changed health care system in Vietnam]. AB - OBJECTIVE: To determine how and where malaria was diagnosed in a forestry area in South-Vietnam and how it was treated. DESIGN: Descriptive. SETTING: Hieu Liem, Dong Nai province, Vietnam. METHOD: In the government hospital and health posts malaria diagnosis and treatment were free of charge while treatment had to be paid for in four private clinics. A population survey was carried out in the forestry area and outside this area: the people were examined for splenic enlargement and a blood sample was analysed. RESULTS: Most patients went to private clinics and it was here that malaria was most frequently diagnosed. In 7.5% of the population in the forest area parasites were found while 1.8% of those living outside the forest appeared to have parasites in the blood. None of the persons with parasitaemia had splenomegaly. Splenomegaly was found in 2.9% of the population, 6.7% in and 0.9% outside the forest area. CONCLUSION: Recent changes in the health sector in Vietnam have liberalized malaria treatment, possibly control. The wide distribution and extensive use of effective drugs like artesunate and mefloquine have probably contributed to reduction of (severe) malaria, but development of resistance to these drugs is to be feared. Control of drug distribution and of prescription practices is urgently needed. PMID- 7477533 TI - [Cardiac myxoma in 13 patients]. AB - OBJECTIVE: To gain insight into the occurrence, the manifestation and the treatment of cardiac myxoma. DESIGN: Retrospective. SETTING: The departments of Cardiology and Cardiac Surgery of Middelheim General Hospital, Antwerp, Belgium. PATIENTS AND RESULTS: Thirteen patients in the period 1985-1995 were subjected to surgical resection of a cardiac myxoma. In accordance with earlier studies, most patients were middle-aged and female. The site of predilection was the left atrium. Cardiac auscultation suggested mitral valve stenosis and insufficiency in three and five patients, respectively; in only one case was the specific 'tumour plop' heard. Cardiac symptoms were atypical thoracic pain (4 times), syncopes (3) and effort dyspnoea (7). Although embolism is described in the literature in 20 70% of the cases, it occurred in only three patients of this group. General symptoms were observed in only seven patients, as against 90% in earlier studies: they were elevated sedimentation rate in five (one also polycythaemia and leucocytosis) and abnormal fatigue in two (one also anorexia) patients. In all 13, the diagnosis of 'intracardiac mass' was made on the basis of ultrasonography. In six of 11 patients who subsequently underwent coronary roentgenography, neovascularization was visualized. All 13 tumours were resected successfully, after which in ten cases a dacron patch was implanted at the level of the interatrial septum. In addition, two patients underwent coronary bypass surgery. Postoperatively, atrial flutter (two patients), atrial fibrillation (one) and fatal cerebral bleeding (one) were observed. The other patients suffered no recurrences. CONCLUSION: Cardiac myxoma is a rare tumour which in general can be accurately diagnosed on the basis of ultrasonography and which almost always can be cured by surgical excision. PMID- 7477534 TI - [Prevalence of HIV infection among drug users in Zuid-Limburg]. AB - OBJECTIVES: To assess the prevalence of HIV infection among intravenous (IDU) and non-intravenous drug users in Heerlen and Maastricht (Southern Netherlands), to detect subgroups of IDU with a higher risk of HIV infection, and to assess the risk of further spread of HIV. DESIGN: Cross-sectional study. SETTING: Heerlen, Kerkrade, Brunssum and Maastricht, the Netherlands. METHODS: Between August 15 and November 25, 1994, a saliva specimen and a short questionnaire were obtained from 449 drug users (340 IDU) in Heerlen (and environs) and Maastricht. Participants were recruited through methadone care (54%), syringe exchange (16%), a street prostitution project (3%), street recruitment (23%) and other drug users (4%). RESULTS: Of the 340 IDU 33 were infected (prevalence: 10% (95% confidence interval: 7-13)), among the 109 non-IDU no infections were found (0% (0-3)). IDU making use of the syringe exchange had a higher prevalence (odds ratio 3.13 (1.37 7.61)). In logistic regression analysis, this finding could not be explained by selection on more risky injecting. One in five currently injecting IDU reported having used someone else's used needle or syringe in the last 6 months. One in five IDU had a non-drug user as steady sexual partner. In sexual contacts between steady partners condom use was low. On the basis of self-reported serostatus it appeared that some infections have taken place in the last two years. CONCLUSIONS: The prevalence of HIV infections among IDU in Heerlen and Maastricht is about 10%. IDU using the syringe exchange have a higher prevalence. This means this prevention reaches the high-risk group, but probably can not avoid all infections. The risk of further spread among IDU is high. The risk of spread to non-IDU and non-drug users is present. PMID- 7477535 TI - [Startle disease: growing rigid with fear]. AB - Hyperekplexia or startle disease was diagnosed in four patients, a girl of 14 months, two men of 45 and 61 years old, and a woman of 56 years old. This is an autosomal dominant inherited disorder, characterized by excessive startle reactions, notably to auditory stimuli. The four patients belong to a Dutch family in which two forms of the disorder occur. The first one is the major form, in which the startle reaction is followed by a temporary generalized stiffening of the body. These patients experience continuous generalized stiffness from birth to about the second year of life. They fall down frequently. The other is the minor form, characterized by an excessive startle reaction only. PMID- 7477536 TI - [Pelvic pain and pregnancy]. PMID- 7477537 TI - [Pelvic pain caused by pregnancy; a new disease?]. PMID- 7477538 TI - [Diagnosis of bronchial asthma and chronic obstructive lung disease by the family physician. Ad hoc consensus group 'Diagnosis of asthma/chronic obstructive lung disease by the family physician']. PMID- 7477539 TI - [Minor symptoms in family practice; chloasma or 'pregnancy mask']. PMID- 7477540 TI - [Prevalence of chronic diseases in the elderly; the ERGO study (Erasmus Rotterdam Health and the Elderly)]. AB - In the Rotterdam Study, prevalence and determinants of chronic diseases in the elderly (age > or = 55 years), were investigated in inhabitants of Ommoord, a suburb of Rotterdam. The study focused on cardiac diseases (myocardial infarction, angina pectoris, cardiovascular risk factors), glaucoma, macular degeneration, osteoporosis, osteoarthrosis and invalidity, dementia (Alzheimer's disease, vascular dementia, Parkinson's disease), epilepsy, cerebrovascular accident. The number of participants was 7983 (3105 men, 4878 women), a response of 78%. The participants were interviewed and were twice examined in an out patient clinic. The results will be described in subsequent issues of this journal. PMID- 7477541 TI - [Coronary heart disease in the elderly; the ERGO study (Erasmus Rotterdam Health and the Elderly)]. AB - OBJECTIVE: To determine the prevalence of cardiovascular disease among elderly people and to investigate the use of cardiovascular medication in this group. DESIGN: Cross-sectional study. SETTING: A quarter in Rotterdam, the Netherlands. METHODS: As a part of the 'Rotterdam elderly study', the cardiovascular anamnesis of 7.983 responding persons > or = 55 years (3105 men and 4878 women) was obtained by means of a questionnaire. RESULTS: Twelve and 4% of men and women, respectively, had at some time had a myocardial infarction, and 5 and 4% a stroke. Almost 7% of both sexes had anamnestic symptoms of angina pectoris. Coronary bypass surgery and percutaneous transluminal coronary angioplasty were done in 4% and 1% of men and 1% and 0.5% of women, respectively. Of all participants 43% used at least one drug for cardiovascular purposes. Half of these used two or more, and a quarter used three or more drugs. Among women, diuretics were the most commonly used drug for cardiovascular purposes (21%), whereas among men beta-blocking agents were most common (15%). CONCLUSION: Cardiovascular diseases are common among elderly men and women. Many of them take at least one drug for cardiovascular purposes. PMID- 7477543 TI - [From the library of the Nederlands Tijdschrift voor Geneeskunde: the legacy of Frederik Ruysch (1638-1731)]. PMID- 7477542 TI - [Risk factors for cardiovascular diseases in the elderly; the ERGO study (Erasmus Rotterdam Health and the Elderly)]. AB - OBJECTIVE: To determine the prevalence of risk factors for cardiovascular disease among elderly people. DESIGN: Prospective cohort study. SETTING: A district of Rotterdam, the Netherlands. METHODS: As a part of the Rotterdam Study information about smoking habits, blood pressure, Quetelet index and serum cholesterol of 7,983 responding persons (78%) (3,105 men and 4,878 women) of 55 years and older was obtained by an interview and physical examination during two visits to a research center. RESULTS: Among men and women there were 29.7% and 16.7% smokers and 60.1% and 25.9% ex-smokers, respectively. Among men the proportion of smokers decreased from 31.0% in the age category 55-59 years to 15.9% in de category > or = 85 years, among women from 28.0% to 2.7%. Systolic blood pressure increased with age in both sexes, while diastolic blood pressure hardly changed. Hypertension (systolic blood pressure 160 mmHg and/or diastolic blood pressure > or = 95 mmHg at a single measurement and/or use of antihypertensive drugs) occurred in 23.3% of men and 28.0% of women. In men, total serum cholesterol decreased gradually with age, whereas in women there was a slight increase up to the category 70-74 years. No evident change in HDL cholesterol with increasing age was observed in men, but in women a decrease was observed until the same level was reached as in men. Thirty-five per cent of men and 49.5% of women had an elevated level of serum cholesterol (> or = 6.5 mmol/l). The prevalence of obesity (Quetelet index > 25 kg/m2) was about 50% in both men and women but was less in the categories from 75 years onwards. Approximately 80% of men and women had at least one risk factor, while in almost half of them two or more risk factors were found. CONCLUSION: Risk factors for cardiovascular disease are common among elderly people. PMID- 7477544 TI - [Disease clusters, 'your own backyard' and demographic synchronization]. PMID- 7477545 TI - [Progressive supranuclear paralysis; a late diagnosis]. PMID- 7477546 TI - [Various unusual complications of intravenous drug use]. PMID- 7477547 TI - [Terminal ileitis in childhood: Crohn disease or gastrointestinal tuberculosis?]. PMID- 7477548 TI - [Is there a connection between alcoholism and the chance of traffic accidents?]. PMID- 7477549 TI - [Diastolic dysfunction of the left ventricle as a cause of heart failure: a clinical problem]. PMID- 7477550 TI - [Study of muscular strength in clinical practice]. PMID- 7477551 TI - [Once more the BCG controversy: no reason for vaccination in the Dutch health care system]. PMID- 7477552 TI - [Surgical treatment of lymphadenitis caused by non-tuberculous mycobacteria in children]. AB - A retrospective study was made of 43 consecutive cases of children with the diagnosis of non-tuberculous mycobacterial lymphadenitis treated in our surgical department between 1976 and 1992. In 16 of the 42 children the diagnosis was initially missed and these children elsewhere underwent incision and drainage resulting in a chronic sinus. In 37 children the disease was localized in the head-and-neck area, while in the other 6 inguinal or axillary lymph nodes were involved. All 43 children underwent a regional lymph node dissection with a partial parotidectomy in three. In 5 children recurrent disease developed within 4 months after dissection. The recurrences were all successfully treated with additional surgery. In none of the children did the dissection result in permanent damage of nerves, especially not of the facial nerve. The cosmetic results were good but depended on the possibilities of entering the neck through a cosmetically ideal incision. Early recognition of the disease is of the utmost importance for an adequate surgical and cosmetically desirable result. An aggressive surgical therapy is advocated and will not result in permanent damage provided the surgeon has the technical skill required for neck dissections. PMID- 7477553 TI - [Surgical treatment of life-threatening mediastinitis following esophageal perforation or leakage after esophagus surgery]. AB - OBJECTIVE: Evaluation of the surgical treatment of life-threatening mediastinitis following oesophageal perforation or leakage after resection for oesophageal cancer. DESIGN: Descriptive retrospective. SETTING: Department of Surgery, University Hospital Utrecht. METHODS: Between June 1989 and October 1994, 18 patients with severe mediastinitis and sepsis following perforation of the oesophagus or leakage after oesophageal resection for oesophageal cancer were treated with aggressive surgery. (Peri-)anastomotic leakage was the cause of mediastinitis and sepsis in 14 patients, 3 patients perforated during endoscopy and 1 patient suffered a spontaneous perforation. Aggressive surgery consisted of removing the oesophageal replacement from the mediastinum, creating an oesophago cutaneostomy and a feeding enterostomy and drainage of the mediastinum and interpleural spaces. RESULTS: All patients survived; however, postoperative morbidity was high. The alimentary tract was reconstructed in 15 patients. CONCLUSION: Aggressive surgical treatment of life-threatening mediastinitis and sepsis as described here can save severely ill patients. Because of the high postoperative morbidity level only patients with perforations of the oesophagus and early stages of oesophageal cancer should be operated. PMID- 7477554 TI - [The work load of family physicians in deprived neighborhoods in Amsterdam: do their patients com more often?]. AB - OBJECTIVE: To determine whether residents from deprived Amsterdam neighbourhoods report more contacts with their general practitioner (GP) than other residents. DESIGN: Face-to-face health interview survey. SETTING: Amsterdam. METHODS: Data on two previous months' contacts with GPs and on background characteristics came from a survey among Amsterdam residents aged 16 years and above (n = 5,121; response: 61.4%). Deprived neighbourhoods were defined as having low mean incomes and high unemployment rates. RESULTS: Residents with a private health insurance from deprived neighbourhoods contacted their GPs significantly less frequently. The contact frequency of persons with compulsory health insurance depended on the definition of deprived neighbourhood: with the preferred definition they contacted their GPs more frequently (0.31/year, 0.49 adjusted for age and gender). This higher contact rate was partially connected with family composition, country of birth, occupational status and chronic psychiatric complaints. CONCLUSION: Quantitatively, residents from deprived Amsterdam neighbourhoods contribute relatively little to the higher workload of Amsterdam GPs; this study gave no information on qualitative aspects. However, Amsterdam residents with compulsory health insurance appeared to visit their GPs more frequently than the Dutch average, and those from deprived neighbourhoods even 9% more. PMID- 7477556 TI - [Extrapulmonary tuberculosis in 3 Dutch patients]. AB - In three patients, two women aged 71 and 59 years and a man aged 49 who had been living in the Netherlands for a long time and who were admitted because of vague symptoms, extrapulmonary manifestations of tuberculosis were diagnosed: tuberculosis of the lumbar spine with psoas abscess, tuberculous peritonitis and adrenal tuberculosis with Addison's disease in a patient with open pulmonary tuberculosis. All three recovered with tuberculostatic therapy (isoniazid, streptomycin, pyrazinamide and rifampicin). PMID- 7477555 TI - [Osteomyelitis as a complication following BCG vaccination]. AB - Osteomyelitis of the proximal femur was diagnosed in a 5-month-old girl after BCG vaccination at the age of 2 weeks. The diagnosis was confirmed by biochemical and molecular techniques (DNA fingerprinting). The girl responded well to antituberculous treatment. PMID- 7477557 TI - [Family physicians mind your affairs!]. PMID- 7477558 TI - [The preconception consultation: a good idea?]. PMID- 7477559 TI - [The skin's immune system]. PMID- 7477560 TI - [Treatment of acute ethmoiditis according to a protocol based on clinical stage]. PMID- 7477561 TI - [Favorable results with liver resection for colorectal liver metastases]. PMID- 7477562 TI - [The serotonin syndrome; a misdiagnosed complication of antidepressive agents]. PMID- 7477563 TI - [Fewer unhealthy years: a utopia?]. PMID- 7477564 TI - [The value of diagnostic tests in infertility]. PMID- 7477565 TI - [Serotonin and cardiovascular diseases]. PMID- 7477567 TI - [Variations in fertility studies in The Netherlands]. AB - OBJECTIVE: To determine how a standard infertility investigation is conducted and to determine the use of fertility tests used in such investigations in relation to WHO guidelines. DESIGN: Questionnaire survey. SETTING: Dutch medical schools. METHOD: A questionnaire survey among the heads of all 8 university and 20 non university teaching departments of obstetrics and gynaecology or their fertility units was conducted. One non-university department failed to respond. RESULTS: The examinations recommended by the WHO (general physical examination, andrological and gynaecological examination, semen analysis, ovulation detection, tubal patency testing) in general were carried out, but general physical examination of the male as a rule was only carried out if indicated. Popular routine examinations not recommended by the WHO were the postcoital test and hysterosalpingography. Regarding the postcoital test, there was variation in the time interval after coitus and the standards for spermatozoal motility. CONCLUSION: Standard fertility investigations are usually based on empiricism and tradition, including the testing recommended by the WHO. PMID- 7477566 TI - [Serotonin in neurology and psychiatry]. PMID- 7477568 TI - [Vertebral deformities and bone density in the elderly; the ERGO Study (Erasmus Rotterdam Health and the Elderly). ERGO Study Group]. AB - OBJECTIVE: To assess the prevalence of vertebral deformities in men and women aged 55 years and over. DESIGN: Cross-sectional study. SETTING: A quarter in Rotterdam, The Netherlands. METHODS: As part of the 'Rotterdam elderly study', 750 responding men and 750 women underwent lateral radiography of the spine and subsequent morphometry of the vertebrae to detect presence of vertebral deformities. Grade 1 and grade 2 deformities were scored on the basis of the results of the morphometry. The entire vertebral column was classified as normal (no deformities), moderate spinal deformity (grade 1 deformities) or severe spinal deformity (grade 2 deformities). Dual Energy X-ray Absorptiometry (DXA) was used to assess the bone mineral density (BMD) at the femoral neck. RESULTS: The prevalence of moderate or severe spinal deformity was estimated to be 18% in men (12% moderate and 6% severe) and 22% in women (11% moderate and 11% severe) after adjustment for the present age distribution of the Dutch population. The prevalence of deformities increased with age, in particular for severe deformities in women aged 70 years and over. A decreasing BMD was observed with increasing severity of vertebral deformities. Men with severe spinal deformity had a 4% lower BMD whereas women with severe spinal deformity had a 6% lower BMD. CONCLUSION: Vertebral deformities are frequently present in the general population, mostly in the higher age groups. There was an association with decreased BMD. In view of the ongoing aging of the population, a future increase in the prevalence of vertebral deformities is to be expected. PMID- 7477569 TI - [Locomotor disability in the elderly; the ERGO Study (Erasmus Rotterdam Health and the Elderly). ERGO Study Group]. AB - OBJECTIVE: Determination of the prevalence of locomotor disability and its association with signs and symptoms of the hips and knees in a Dutch general population aged 55 years and over. DESIGN: Cross-sectional population-based study. SETTING: Rotterdam-Ommoord, the Netherlands. METHOD: During home interviews of 1,901 men and 3,132 women aged 55 years and over living in the Ommoord district of Rotterdam, locomotor disability was assessed using the Health Assessment Questionnaire (HAQ). Morning stiffness and pain in the hips and knees during the past month were assessed. At the research centre weightbearing radiographs of the hips and knees were taken. Radiological osteoarthritis was defined as Kellgren score 2 or more. A complete physical examination of the hips and knees was performed. RESULTS: The prevalence of locomotor disability was 21.9% for independently living men and 36.0% for independently living women. In homes for the elderly these figures were 84.0% en 96.5%, respectively. There were significantly elevated age-adjusted odds ratios for locomotor disability for women, people in homes for the elderly, independently living men with only primary education, independently living participants with a below-median net annual income and widowed men in homes for the elderly. Taking the associations between the independent variables into account only morning stiffness, pain in the hips and knees and restricted flexion of the hips and knees were independently associated with locomotor disability in men. In women radiological osteoarthritis of the hips and knees, restricted endorotation of the hip, instability and valgus deformity of the knees were also associated with disability. CONCLUSION: The prevalence of locomotor disability in people aged 55 years and over in the general population was high and associated with female sex, low education, low income and living in a home for the elderly. Of the signs and symptoms of the hips and knees only pain in the hips and knees, morning stiffness and restricted flexion of the hip were independently associated with locomotor disability. Radiological osteoarthritis of the hip and knee did not contribute much to the explanation of locomotor disability. PMID- 7477571 TI - [Listening and observing]. PMID- 7477570 TI - [African sleeping sickness in The Netherlands]. AB - Of the four most dangerous protozoal infections acquired in (sub)tropical regions, falciparum malaria, amoebic abscess of the liver, visceral leishmaniasis (kala azar) and African trypanosomiasis (sleeping sickness) only the fourth was up to now unreported in the Dutch medical literature. Two case histories are presented: a Cameroonian woman, resident in the Netherlands for two years, suffering from West African type sleeping sickness, and a Dutch tourist who acquired East African trypanosomiasis while travelling through Zimbabwe. Although the parasites are morphologically identical, clinical and epidemiological characteristics are distinctly different. The West African type, rarely if ever observed in Europeans, has an insidious chronic course leading to the features of classical sleeping sickness. Differential diagnosis is difficult. The East African variety runs an acute course in Europeans leading to death within days due to myocarditis. It is therefore mandatory for the diagnosis to be made as soon as possible in order to initiate specific therapy. Both patients recovered after specific therapy. PMID- 7477572 TI - [Treatment of a keloid of the earlobe]. PMID- 7477573 TI - [Clinical study of adjuvant systemic chemotherapy in breast cancer in The Netherlands assumes a greater number of patients than is available]. PMID- 7477574 TI - [Orbital cellulitis as a sequela of paranasal sinusitis]. PMID- 7477575 TI - [Influenza vaccine for all the elderly!]. PMID- 7477576 TI - [Current developments in the area of maturity onset diabetes of the young (MODY)]. PMID- 7477578 TI - [Current developments in the therapy of the myelodysplastic syndrome]. PMID- 7477577 TI - [Myelodysplastic syndrome; current insights in pathogenesis and treatment]. PMID- 7477579 TI - [Influenza vaccine in 85-and-over aged: motivation of elderly and family physicians to vaccinate or not]. AB - OBJECTIVE: To determine on what grounds persons over 85 years are or are not vaccinated against influenza. DESIGN: Descriptive. SETTING: Gerontological Research Centre and Department of General Practice Medicine, University of Leiden, the Netherlands. METHOD: A random sample (n = 331) of the general population of Leiden aged 85 years or older and not institutionalized were interviewed. Early in October 1993, 163 interviews were suitable for analysis (response rate 54%). An analysis of non-responders revealed no significant difference as regards gender and housing, but persons aged 90 and older were under-represented. The general practitioners (n = 41) of the 163 elderly persons were interviewed about these persons (response: n = 127; 78%) and about influenza vaccination in general (response: n = 33; 82%). RESULTS: The vaccination coverage rates were 51% according to the elderly and 56% according to the GPs; those of elderly people with an indication (48% according to themselves and 64% according to the GP) 52% and 67%, respectively. Reasons for elderly people not to accept vaccination were that they considered vaccination unnecessary, that they felt well and wanted to avoid possible adverse effects. A doctor's recommendation to have vaccination done was a positive influence. GPs' motives not to vaccinate were absence of an indication and the elderly person's wish. CONCLUSION: Only two thirds of those over 85 not living in a nursing home with an indication for influenza vaccination were indeed vaccinated. Information about the possible damage done by influenza and about the pros and cons of vaccination, together with a doctor's advice to be vaccinated appeared to have a positive effect on the vaccination coverage. PMID- 7477581 TI - [Myelopathy following influenza vaccination]. AB - A healthy woman aged 56 was vaccinated for the second year in succession against influenza. Three days afterwards she developed a strange sensation in the legs and lower abdomen, accompanied by difficulties of urinating and defaecation. Motor strength and reflexes were normal at neurological examination. There was, however, an altered sensibility below the dermatome T7. Additional investigations were all normal, and the diagnosis of myelopathy after influenza vaccination was made. Over six months there was gradual amelioration. PMID- 7477580 TI - [Increase of degree of vaccination against influenza in at-risk patients by directed primary care invitation]. AB - OBJECTIVE: To increase the coverage of influenza vaccination by a programmatic approach in a large population, aimed at the general practitioners (GPs). DESIGN: Intervention study (prospective). SETTING: The region of central Brabant (Tilburg and some surrounding communities), the Netherlands. METHOD: The intervention was carried out in a population of 133,000 people (registered at 42 offices of 56 GPs, and at 27 pharmacies). Medication data from pharmacists were used for the selection of patients at risk by the GPs. RESULTS: About 15,000 patients were selected by the GPs and invited for vaccination. The (final) coverage was 75.5% in 1993, a 56% increase over 1992. The increase in the group of comparable but non-participating GPs was 18%, in the Netherlands as a whole it was 8%. CONCLUSION: It is quite possible to achieve a considerable increase of the coverage of influenza vaccination. PMID- 7477582 TI - [Influenza in the 1994/95 season; composition of vaccine for the 1995/96 season]. AB - The 1994/'95 season in the Netherlands was marked by a limited influenza activity which only emerged in late February. The influenza activity remained elevated until the end of April, which is unusually late, and epidemic activity was only reported in the south of the country. Both influenza A/H3N2 and B viruses were isolated in this period. In addition, influenza A/HINI viruses were isolated for the first time since March 1993, from two patients. The majority of the influenza A strains that circulated in the Netherlands in 1994/'95 reacted well with ferret antiserum raised against the strains of the 1994/'95 influenza vaccine, which therefore probably offered good protection. The reactivity of the B strains to antiserum raised against the vaccine strain, B/Panama/45/90, was only moderate, which implies that the protection against the Dutch influenza B strains was not optimal. Based on the results of the worldwide influenza surveillance, the World Health Organization (WHO) has recommended an alteration in both the A/H3N2 and the B component for the vaccine of 1995/1996. PMID- 7477583 TI - [Schistosomiasis after swimming in the lake of Malawi]. PMID- 7477584 TI - [Stereotactic radiosurgery]. PMID- 7477585 TI - [High-resolution computerized tomography of the lungs: current diagnostic possibilities]. PMID- 7477586 TI - [Treatment and prevention of generalized mycoses]. PMID- 7477587 TI - Director of State Health Department optimistic about public-private efforts. Interview by Carol McShane. PMID- 7477588 TI - Restructured Dept. of Health has four divisions, redefines management. PMID- 7477589 TI - The Clarkson debate: for-profit vs not-for-profit. PMID- 7477590 TI - Understanding Medicaid managed care in Nebraska. PMID- 7477591 TI - Testifying before the legislature: a call for action. PMID- 7477592 TI - Nursing involvement in the legislative process: you can make a difference! PMID- 7477593 TI - [Vigabatrin symposium within the congress "Epilepsy Europe", 3 September 1992, Glasgow]. PMID- 7477594 TI - [Perazine therapy. Seminar, Hamburg, 12-14 November 1992]. PMID- 7477595 TI - [Is there a new trend in antidepressive therapy? Bonn, 2 July 1993]. PMID- 7477596 TI - [Lamotrigin--a new anticonvulsant for epilepsy. Conference, 25 June 1993, Frankfurt]. PMID- 7477597 TI - [Therapeutic use of neuroleptics. Satellite symposium "Risperdal: new horizons for the schizophrenic patient". 9th World Congress for Psychiatry, Rio de Janeiro, 6-12 June 1993]. PMID- 7477598 TI - [Vitamin B in human metabolism. Diabetic polyneuropathies. 3rd Hohenheimer symposium. November 11, 1992. Alcoholic neuropathies. Stuttgart, 3 June 1992]. PMID- 7477599 TI - [Diagnosis and therapy of dementia syndrome. Italo-German symposium on primary and vascular dementia. 26-27 March 1993]. PMID- 7477600 TI - [Stroke: new perspectives in prevention. Scientific symposium, 24 April 1993, Frankfurt]. PMID- 7477601 TI - [Anxiety--compulsiveness--depression. Symposium, Mainz, 8 May 1993]. PMID- 7477602 TI - [Psychiatrically relevant side effects of non-psychopharmacological pharmacotherapy]. AB - We briefly review the possibilities of side effects caused by therapy with drugs other than psychotropic agents. These complications are mainly depressive/manic, paranoid-hallucinatoric/deliriant or anxiety/panic syndromes. Especially elderly patients and patients with multiple diseases are the preferred group with these side effects, because they are most likely to show a disturbance of blood-brain barrier and renal clearance. We will discuss the psychiatrically meaningful side effects of frequently used drugs, and we will present a concept of how to reduce such unwanted side effects. PMID- 7477603 TI - [Clinical aspects and follow-up of dopamine-induced psychoses in continuous dopaminergic therapy and their implications for the dopamine hypothesis of schizophrenic symptoms]. AB - We present the case reports of 11 Parkinsonian patients who developed acute psychosis under continuous dopaminergic stimulation. Psychopathologically, two of the patients mainly suffered from organic hallucinosis, while nine patients showed the clinical symptoms of delirium. The clinical course and psychopathological findings in these patients did not differ from other acute organic psychoses. However, the symptoms of these dopaminergically induced psychoses varied significantly from the psychopathological findings of paranoid schizophrenic patients who were regularly treated and evaluated in our clinic. These differences in symptoms and clinical course of dopamine-induced and schizophrenic psychosis do not support the hypothesis that the pathogenesis of schizophrenic symptoms can be explained only by a hyperfunction of dopaminergic transmission. Instead, the involvement of other neurotransmitter system must be considered in order to explain the pathogenesis of schizophrenic symptoms on a neurobiological level. PMID- 7477604 TI - [Psychiatric disorders in hospitalized internal medicine and surgical patients. Prevalence and need for treatment]. AB - The prevalence of psychiatric disorders in general hospital inpatients can be estimated at 30-50%. In Germany, hardly any studies have provided an assessment of prevalence rates and specific treatment necessities. In this study, 400 patients from medical and surgical departments were interviewed with a structured clinical interview. The point prevalence of all psychiatric disorders (clinical diagnoses) was 46.5%. The most prevalent groups of disorders were organic brain syndromes, depressive disorders, and alcoholism. With regard to disease entities, no significant differences between the samples from internal medicine and from surgery could be found. Compared to a German population sample, only organic brain syndromes, depressive reactions and alcohol dependency tend to be higher in a general hospital. Consultation/liaison interventions were found to be necessary in nearly half of the whole sample. In one-fourth of the patients continuous psychiatric care after hospital discharge was required. PMID- 7477605 TI - [Effects of psychopharmacologic therapy on heart rate variation]. AB - Twenty patients suffering from schizophrenia and 36 patients suffering from endogenous depression underwent a standardized heart rate analysis before drug therapy. The patient's parameters of heart rate variability (HRV), which are controlled by the parasympathetic nervous system and which are independent of heart rate, did not significantly differ from the HRV parameters of normal control subjects. Ten of the patients with schizophrenia were treated with 200 400 mg of clozapine/day as monotherapy, while the other ten patients received a combination of different psychotropic drugs. The depressed patients were either treated with 150 mg of amitriptyline/d (n = 24) or 20 mg of paroxetine/d (n = 12) as monotherapy, respectively. After treatment with an average of 300 mg of clozapine/d for 4 weeks or with 150 mg of amitriptyline/day for 2 weeks, all of the patients HRV parameters had significantly decreased (P < 0.001). At this time, about 90% of these patients fulfilled the criteria of cardiovascular autonomic neuropathy. However, treatment with 20 mg of paroxetine/day for 2 weeks had no impact on any of the heart rate parameters. Under amitriptyline treatment, HRV parameters were found to correlate significantly with the plasma levels of amitriptyline/nortriptyline in a group of 104 depressed patients. Thus, determination of decreased HRV parameters is suggested to be a useful tool for the detection of overdosage with amitriptyline. It has not yet been elucidated whether or not the observed HRV decrease, which is probably at least in part due to the anticholinergic side effects of clozapine and amitriptyline, has any impact on patient health.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7477606 TI - [Chronification of sleep disorders. Results of a representative survey in West Germany]. AB - In a representative selection of German citizens who were older than 13 years of age, 1,997 were asked about their sleep complaints. They were also asked how frequently a physician was consulted and how often sleeping pills were taken. According to our results sleep disturbances are an important health problem in Germany. Every fourth person suffers at least sometimes from difficulties in falling asleep and/or staying asleep, problems which are not due to external influences. Seven percent suffer frequently or always from these complaints; 15% report that they are frequently tired or that they are always tired during the day. Ten percent of all persons suffering from sleep complaints take sleeping pills daily or at least sometimes during the week. Forty-five percent of all persons who take hypnotics daily still frequently or always suffer from difficulties in falling asleep and/or staying asleep. Furthermore, sleep complaints tend to become chronic: 75% of the sleep-disturbed population are chronically ill, having had complaints for more than 2 years of since childhood. They suffer from sleep disturbances and from reduced performance during the following day. Nevertheless, it appears as though neither patients nor physicians take insomnia seriously. This might answer the question of why only 17% of the persons who do not suffer more than 2 years from sleep disturbances and only 49% of the chronically ill population consult a doctor because of their sleep disturbances. The results of this study indicate the importance of informing patients and physicians about insomnia and different ways of treating it. PMID- 7477607 TI - [Premorbid intelligence diagnosis with the MWT-B(Multiple Choice Word Test-B) in patients with Alzheimer disease]. AB - The usefulness of the multiple-choice word test (MWT-B) was tested as a measure for premorbid intelligence in 107 patients with probable Alzheimer's disease following the NINCDS-ADRDA criteria. The patients were allocated to three groups according to severity of dementia: mild (n = 40), moderate (n = 41) and severe (n = 26). Dementia severity was assessed with the Mini Mental State Test (MMST). The mean of the raw values of the MWT-B (maximum 37) in the group with mild dementia was 28.0 (SD = 5.57), in the group with moderate dementia 23.3 (SD = 8.01), and in the group with severe dementia 6.7 (SD = 8.44), indicating a parallel worsening with the other tests. The MWT-B value of the whole sample correlated with the MMST value (r = 0.70); a significant correlation still existed in patients with mild and moderate dementia. These results confirm that the MWT-B is relevant for dementia. The rate of cerebral glucose metabolism, measured with positron emission tomography (PET) and 18F-fluorodeoxy-glucose (FDG) correlated significantly with the MWT-B scores in the Alzheimer-sensitive temporoparietal region (r = 0.43; P = 0.001) and frontal brain region (r = 0.37; P < 0.05). Whether or not it can be used as a screening measure of dementia will have to be decided on the basis of its scales for sensitivity and specificity. PMID- 7477608 TI - [Computer-assisted individualized memory training in Alzheimer patients]. AB - Memory training programs for cognitively impaired patients have often been criticized for their lack of relevance to everyday activities. We therefore report our experience with two patients suffering from probable Alzheimer's disease who were trained with a new computer-based program recently developed by our research group. An everyday task of personal relevance to the patient was simulated and trained on a PC touch screen using personal photographs of the patient's surroundings and biography. The patient's training performance (time, mistakes, number of advices) improved substantially. While psychopathometric tests showed no significant effects with regard to general cognitive performance, levels of motivation were high and there was a positive acceptance of the training, signs of emotional activation and of a transfer of the trained skills into real situations. PMID- 7477609 TI - [Willingness to participate in dementia research by relatives of Alzheimer patients and family members of cognitively normal persons]. AB - We investigated the willingness to participate in dementia research of relatives of demented (n = 52) and cognitively unimpaired elderly individuals (n = 42). This unrepresentative sample was given a questionnaire to determine the rate of approval with respect to two research examples, the first of which referred to a therapeutic study with potential individual benefits, the second to a diagnostic research project without such benefits. Independent of potential individual benefits, an overwhelming majority acknowledged the necessity for dementia research and consented to participate in the given research examples. The rate of approval was not related to the experiences of the interviewed persons in the care of relatives suffering from dementia. PMID- 7477610 TI - [Optical and tactile hallucinosis as clinical onset of Creutzfeld-Jakob disease]. AB - A patient suffering from Creutzfeld-Jakob disease (CJD) complained of optical hallucinations, followed by feelings of persecution. Clinical, encephalographic and neuropathological findings pointed towards a focalized beginning of the disease process in the right frontal and temporal lobe. The hypothesized further spread of the infectious agent, as indicated by electroencephalographic and neuropathological findings, was accompanied by periacute manifestation of dementia and an irreversible disturbance of vigilance. The diagnosis of CJE can be especially difficult in the early stages of the disease, with dementia, myocloni and typical electroencephalographic findings still absent. PMID- 7477611 TI - [What is new in biologically oriented psychiatry research? Congress report of the 6th Congress of the German Society of Biological Psychiatry, 6-8 October 1994 in Munich]. PMID- 7477612 TI - [Folie a deux]. PMID- 7477613 TI - Hypercoagulability, intraglomerular coagulation, and thromboembolism in nephrotic syndrome. PMID- 7477614 TI - Evolution of serum erythropoietin after androgen administration to hemodialysis patients: a prospective study. AB - A prospective study of the evolution of serum erythropoietin level after androgen therapy was carried out in a group of 25 male patients on chronic hemodialysis treatment with nonferropenic anemia (serum ferritin > 50 ng/ml). The androgen used was nandrolone decanoate (200 mg/week intramuscularly, for 6 months). There was an increase of serum erythropoietin, that reached statistical significance in the 2nd week of treatment (8.6 +/- 6.4 vs. 14.2 +/- 9.8 mIU/ml, p < 0.05), and a stabilization after 1 month (1 month: 17.8 +/- 11.2 mIU/ml, 6 months: 19.6 +/- 14.9 mIU/ml). The hemoglobin also experienced a parallel increase to that observed in serum erythropoietin (basal value: 8 +/- 0.9 g/dl; at 1 month postandrogen: 9.2 +/- 1.3 g/dl, p < 0.001; at 6 months: 10.7 +/- 1.8 g/dl, p < 0.001). According to their response of serum erythropoietin the patients were divided into responders (15 patients) and nonresponders (10 patients). There were no differences between them concerning age, basal levels of serum erythropoietin and hemoglobin, and dose of nandrolone decanoate in relation to body weight. The evolution of hemoglobin was similar in both groups, and a correlation between serum erythropoietin and hemoglobin was not observed in the responder group. Fourteen patients were studied after androgen was discontinued. The serum erythropoietin returned to basal levels 6 weeks after the last dose of nandrolone decanoate (7.7 +/- 5.4 mIU/ml). However, hemoglobin was above the basal levels 16 weeks after discontinuing androgen (9.5 +/- 1.1 g/dl, p < 0.05), with no differences between the responder and nonresponder group.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7477615 TI - Long-term control of hyperparathyroidism in advanced renal failure by low phosphorus low-protein diet supplemented with calcium (without changes in plasma calcitriol). AB - Phosphorus (Pi) retention linked to chronic renal failure (CRF) favors secondary hyperparathyroidism (HPT). Reduction of Pi and protein intake has been shown to prevent the development of HPT in CRF. The aim of the present study was to assess in patients with advanced CRF the long-term effects on phosphate and calcium metabolism of a low-Pi (5-7 mg/kg/day), low-protein (0.4 g/kg/day) diet providing 300 mg/day calcium (Ca) and supplemented with amino acids and ketoacids, Ca carbonate (400-800 mg/day) and vitamin D2 (1,000 IU/day). Twenty-nine patients with advanced CRF (glomerular filtration rate (GFR) 13.7 +/- 4.5 ml/min) were selected for the study, on the basis of a follow-up of a least 2 years and a satisfactory compliance to the prescribed diet. At the start of the study, biological evidence of HPT was present with increased plasma PTH concentration (144 +/- 95 pg/ml), increased plasma Pi (1.57 +/- 0.33 mmol/l), an increase in alkaline phosphatase activity and plasma osteocalcin concentration. Plasma PTH concentration was positively correlated with plasma Pi and inversely with plasma Ca concentrations and GFR. Pi and protein restriction induced a significant correction of HPT within 3 months after starting the diet. After 2 years of diet, despite the diminution of GFR (11.1 +/- 3.7 ml/min, p < 0.0001), plasma PTH was still lower than at the start of the diet (88 +/- 57 pg/ml, p < 0.01), as was plasma Pi (1.32 +/- 0.24 mmol/l, p < 0.001), total plasma Ca being higher (p < 0.01). Plasma PTH levels were correlated only to plasma Ca concentrations.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7477617 TI - Prevention of hepatitis C virus in dialysis units. AB - To understand how to prevent the diffusion of hepatitis C virus (HCV) in dialysis units, 289 chronic dialysis patients treated in a renal department from the beginning of 1990 to June 30, 1993, were studied. Patients were screened monthly for alanine aminotransferase values and every 3 months for anti-HCV antibodies. At the beginning of the study the prevalence of anti-HCV antibodies was 24.7%. Two study groups were defined. In the first, anti-HCV-positive patients were treated on separate machines; in the second, 13 anti-HCV-positive and 13 negative patients shared the same machines. Patients in the study were treated with traditional dialysis, employing low-permeability membranes and disposable dialysate circuits on machines without an ultrafiltration control device. The 'universal precautions' were rigorously applied. The use of blood transfusions was markedly reduced. Although new patients starting dialysis treatment revealed a high frequency of HCV positivity (10.8%), the overall prevalence of HCV infection in the department did not increase during the follow-up period. Furthermore, no seroconversion was found in patients on dialysis treatment, not only in the section where anti-HCV-positive patients were treated on separate machines, but also in the section where anti-HCV-positive and anti-HCV-negative patients shared the same machines. The possibility of an intradialytic diffusion of HCV appeared to be very low and the treatment of infected patients on separate machines not strictly necessary. PMID- 7477616 TI - Gastroparesis in diabetics on chronic dialysis: clinical and laboratory associations and predictive features. AB - Clinical and laboratory features and risk factors for diabetic gastroparesis (DGP) were investigated in 226 diabetics on chronic dialysis; 106 subjects (43%) had DGP diagnosed by persistent vomiting improved with the use of prokinetic agents and 120 (control group) had no clinical DGP. Type 1 diabetics had DGP more frequently than type 2 diabetics (70 vs. 37%). The DGP group had longer duration of diabetes (21 +/- 8 vs. 13 +/- 6 years), higher frequency of diabetic orthostatic hypotension (95 vs. 33%), enteropathy (49 vs. 5%), blindness (52 vs. 23%), myocardial infarction (86 vs. 42%), extremity gangrene (54 vs. 27%) and cerebrovascular accidents (43 vs. 25%), lower serum albumin 32.3 +/- 3.9 vs. 35.4 +/- 3.8 g/l), urea (24.0 +/- 5.5 vs. 25.5 +/- 5.5 mmol/l) and creatinine (710 +/- 210 vs. 820 +/- 220 mumol/l), and higher serum TCO2 (20.9 +/- 3.1 vs. 19.8 +/- 2.7 mmol/l) than the control group (all differences significant at p +/- 0.004). Glycemic control was adequate in 24% of the DGP group subjects and 83% of the control subjects (p < 0.001). Annual hospitalization rate was 49 +/- 48 days/patient in the DGP group and 16 +/- 27 days/patient in the control group (p < 0.001). Median patient survival was 24 +/- 2 months in the DGP group and 61 +/- 9 months in the control group (p < 0.0001). Logistic regression identified long duration of diabetes and poor glycemic control as risk factors for DGP. In diabetics on dialysis, DGP is associated with high frequency of other diabetic complications, low serum albumin and creatinine, and high morbidity and mortality.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7477618 TI - Impairment of tubular secretion of urate in renal transplant patients on cyclosporine. AB - The prevalence of hyperuricemia was investigated in 214 kidney allograft recipients, 81 were on azathioprine and steroids and 133 on cyclosprine (CyA) and low-dose steroids or on triple therapy. All had stable renal function, serum creatinine < 2.5 mg/dl, and a follow-up between 12 and 120 months. At the time of the study, blood and urine samples were obtained to perform tests of renal function. The renal handling of urate was evaluated by a combined pyrazinamide and probenecid test in 35 selected patients (12 normouricemic on azathioprine, 9 normouricemic on CyA and 14 hyperuricemic on CyA). The prevalence of hyperuricemia was higher in the group of patients on CyA (19.7 vs. 66.9%, p < 0.001), as well as the concentration of serum urate (6.1 +/- 1.9 vs. 7.6 +/- 1.7, p < 0.001), and serum creatinine (1.2 +/- 0.3 vs. 1.4 +/- 0.4, p < 0.001). In patients on CyA, multivariate analysis showed that the most important predictive variables of hyperuricemia were: serum creatinine, FEurate, diuretic use and CyA blood levels (r = 0.73, p < 0.0001). Thirteen patients on CyA (9.9%) had at least one episode of gouty arthritis. Those patients were older than the hyperuricemic patients without gout (45.7 +/- 6.7 vs. 37.1 +/- 13.5 years, p < 0.01), had worse renal function (serum creatinine 1.9 +/- 0.4 vs. 1.5 +/- 0.4 mg/dl, p < 0.01), and higher prevalence of hypertension (100 vs. 63.1%, p < 0.05).(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7477619 TI - The well-functioning renal graft evaluated by color Doppler flowmetry. AB - The value of color Doppler ultrasound in the renal transplant follow-up has been evaluated. To do so we used a standardized protocol of analysis on a group of 86 outpatients with different transplant ages and a good and stable graft function defined as a plasma creatinine level < 105 microM/l. Renal volume increased after transplantation and averaged 198 +/- 54 cm3. The graft volume was positively related to the transplant age (p = 0.04). Mean renal blood flow/1.73 m2 body surface area was 301 +/- 98 ml/min, a value at the lower limit of normality. A statistical inverse relationship between renal blood flow and transplant age was found (p = 0.04). Renal vascular resistances increased along with the transplant age (p = 0.003). Renal function evaluated by creatinine plasma levels and creatinine clearance values did not show any statistical correlation with color Doppler findings in normal grafts. In conclusion, ultrasound measures allow us to obtain more sensitive information about the graft status and might be used for a better evaluation of the transplant follow-up. PMID- 7477620 TI - Acute renal failure associated with Yersinia pseudotuberculosis infection. AB - Since 1987, we have experienced 11 children with acute renal failure (ARF) associated with Yersinia pseudotuberculosis (YP) infection. The illness began with the sudden onset of high fever, skin rash and gastrointestinal symptoms. Later in the course, periungual desquamation developed, mimicking Kawasaki disease. Elevated erythrocyte sedimentation rate, C-reactive protein and thrombocytosis were noticeable, and mild degrees of proteinuria, glycosuria and sterile pyuria were common. ARF, which typically developed about 1-3 weeks after the onset of fever, underwent a benign course with complete recovery. The renal biopsies mainly revealed findings of acute tubulointerstitial nephritis. YP should be considered as one of the causes of acute tubulointerstitial nephritis causing ARF, especially in children. PMID- 7477621 TI - Serum hepatocyte growth factor levels in patients with chronic renal failure. AB - The serum levels of hepatocyte growth factor (HGF) were determined in chronic renal failure (CRF) patients. Nondialysis patients with renal insufficiency had significantly higher serum HGF than normal subjects (0.34 +/- 0.10 ng/ml, n = 21 vs. 0.19 +/- 0.05 ng/ml, n = 15; p < 0.001), and the elevated serum HGF correlated with their serum creatinine levels. Hemodialysis (HD) patients treated for 5-10 years showed higher serum HGF than those receiving HD for 1 year or less (0.45 +/- 0.14 ng/ml, n = 8 vs. 0.33 +/- 0.11 ng/ml, n = 9; p < 0.05). Continuous ambulatory peritoneal dialysis patients also showed elevated serum HGF levels comparable to those of HD patients. There was no difference in serum HGF levels in HD patients with or without acquired cystic disease of kidney. Consequently, serum HGF is elevated in CRF, which may be attributed to the increased production of HGF in response to the chronic renal injury, the effect of heparin, or reduced removal of serum HGF in CRF patients. PMID- 7477622 TI - Binding capacity of serum IgA to jacalin in patients with IgA nephropathy using jacalin-coated microplates. AB - Binding capacity of serum IgA to jacalin in 22 patients with IgA nephropathy, 14 patients with diffuse mesangial proliferative glomerulonephritis (non-IgA nephropathy) and 20 age-matched healthy adults was examined by enzyme-linked immunoassay (ELISA) using jacalin-coated microplates. In contrast to previous findings, the binding capacity of serum IgA to jacalin in patients with IgA nephropathy measured by ELISA using jacalin-coated microplates was significantly higher than that in healthy adults. The ratio of serum IgA levels measured by this method to those obtained by single radial immunodiffusion was significantly increased in patients with IgA nephropathy. It appeared that the capacity of serum IgA binding to jacalin was marked in these patients. It is concluded that the binding capacity of serum IgA to jacalin is not ubiquitously impaired in all patients with IgA nephropathy. PMID- 7477624 TI - Iron depletion in the remnant kidney. AB - Urinary and proximal tubular iron are increased after subtotal nephrectomy, and iron depletion has been shown to be beneficial in proteinuric models of chronic renal disease in rats. In this study, iron depletion by low iron pair-fed diet and periodic phlebotomy was induced for 6 months in rats with partial (5/6) nephrectomy, resulting in a reduction in hematocrit and serum iron in all iron deficiency subgroups. Tubular iron, assessed by energy dispersive analysis and electron microscopy, was reduced in quantity but not number of iron-containing lysosomes only within 1 subgroup of severe iron deficiency (p < 0.05). There was no improvement in serial isotopic glomerular filtration rate measurements, urinary protein and transferrin excretion, tubular damage scores, serum creatinine, or measures of reactive oxygen species (ROS) generation. In a subgroup of rats with no supplementation of sulfhydryl amino acids (cysteine and methionine) which can act as ROS scavengers, iron deficiency increased urinary protein excretion (213.3 +/- 23.0 mg/24 h, mean +/- SEM, vs. 87.4 +/- 16.1, p < 0.001), urinary transferrin excretion (p < 0.05), kidney weight (p < 0.05) and tissue malondialdehyde, a lipid peroxidation product (0.78 +/- 0.16 nmol/mg protein vs. 0.57 +/- 0.19, p < 0.05), consistent with increased ROS generation. Hence, no beneficial effect of iron-deficiency was demonstrated by any measure of structure of function in the remnant kidney, and it may enhance damage if sulfhydryl repletion is not provided. PMID- 7477623 TI - Secondary hyperparathyroidism and depression in chronic renal failure. AB - This study was aimed at examining the relation between secondary hyperparathyroidism (sHPT) and depressive syndromes in chronic renal failure (CRF). SAMPLE AND METHODS: 59 chronic CRF patients and 16 depressive patients without CRF were included. Parathyroid hormone (PTH), calcium and phosphate as well as psychopathology were measured. RESULTS: Depressive CRF patients with cognitive disorders show higher levels of PTH than less or not depressive subjects. With respect to the literature, our results give reason to assume that sHPT may play a pathogenetic role in those depressive syndromes which are additionally characterized by cognitive disorders. PMID- 7477625 TI - Renal lesion of type Ia glycogen storage disease: the glomerular size and renal localization of apolipoprotein. AB - In order to investigate the glomerular size and renal localization of apolipoprotein in type Ia glycogen storage disease, a renal biopsy was performed in two proteinuric patients. Histopathological examination of the biopsy specimens revealed focal sclerotic glomerular sclerosis in both patients. The mean glomerular area was 21.6 +/- 11.6 x 10(3) microns 2, indicating enlargement of the glomeruli. Immunohistochemical staining of the specimens for apolipoprotein showed localization of apolipoprotein AI on the inner side of the glomerular capillary wall, and in proximal tubular epithelial cells. In one patient with a history of several episodes of hypoglycemia, treatment with corn starch improved the carbohydrate and lipid metabolic profile and reduced the daily urinary protein excretion from 2.23 to 0.5 g. These results suggest that focal sclerotic glomerular lesions associated with type Ia glycogen storage disease may be related to disorders of carbohydrate and lipid metabolism. PMID- 7477626 TI - Glomerulocystic kidney disease in a young adult. AB - We report an 18-year old woman who had glomerulocystic kidney disease (GCKD) without a family history of renal disease or hypertension and no known congenital abnormalities. Her renal function was normal. Renal biopsy showed cystic dilatation of the Bowman's spaces and atrophy of the glomerular tufts. Electron microscopy revealed specific changes in the basement membranes of noncystic glomeruli, suggesting a congenital origin for her renal pathology. This relatively rare case contrasts with the usual presentation of GCKD in neonates or children. PMID- 7477627 TI - Newly discovered familial juvenile gouty nephropathy in a Japanese family. AB - Our attention was initially called to 2 young Japanese sisters with gout and renal insufficiency, which led to an investigation of members of their family with similar conditions. One sister, a 26-year-old woman who had suffered from polyuria since infancy, suffered from gout and renal insufficiency. Her younger sister also had a history of polyuria, hyperuricemia, and moderately reduced renal function. Their urinary uric acid levels were reduced but purine enzyme activities in the erythrocytes were normal. A renal biopsy specimen from the younger sister showed severe interstitial fibrosis with tubular atrophy. An investigation of the family revealed an autosomal dominant transmission pattern. We believe these are new familial cases of juvenile gouty nephropathy found in a Japanese family. PMID- 7477628 TI - Low prevalence of hepatitis C virus antibodies with primary membranous nephropathy and membranoproliferative glomerulonephritis in Hong Kong. PMID- 7477629 TI - Effect of benidipine on decreasing glomerular expansion in the experimental nephrotic syndrome. PMID- 7477630 TI - Cystatin C: a promising marker of glomerular filtration rate to replace creatinine. PMID- 7477631 TI - Treatment of severe acute hypernatremia and renal failure by hemodialysis. PMID- 7477632 TI - Nephrotic syndrome developed in a patient with acute promyelocytic leukemia treated with daunomycin. PMID- 7477633 TI - Decrease of urine protein after treatment with probucol in patients with mild glomerulonephritis and arteriolosclerosis. PMID- 7477634 TI - Early and delayed onset cardiac tamponade secondary to subclavian catheter for dialysis. PMID- 7477635 TI - Effects of gentamicin on iron and copper contents of kidney tissue. PMID- 7477636 TI - Determination of radical species in the kidney of rats with chronic renal failure by the spin trapping method. PMID- 7477637 TI - Spontaneous renal allograft rupture: still a threat. PMID- 7477638 TI - The ghost in the machine can be exorcised. PMID- 7477639 TI - Genetic linkage analysis of autosomal dominant polycystic kidney disease in a Turkish family. PMID- 7477640 TI - Acrodermatitis enteropathica-like syndrome in a dialysis patient. PMID- 7477641 TI - Deoxyspergualin: a new immunosuppressive drug for the treatment of auto-immune disease. PMID- 7477642 TI - Prevalence and clinical course of hepatitis B and hepatitis C liver disease in ciclosporin-treated renal allograft recipients. AB - We performed this study to evaluate prevalence and clinical course of hepatitis B surface antigen (HBsAg)-positive and anti-hepatitis C virus (HCV)-positive renal transplant recipients. HBsAg positivity was 13.7 and anti-HCV positivity 12.8%. Before transplantation, the HBsAg positivity was observed in in 83.5% of the patients, and 16.4% of the patients acquired HBsAg after renal transplantation. In the HCV group, anti-HCV positivity was observed in 47.1% before transplantation, and 19.6% acquired anti-HCV after renal transplantation. The prevalence of chronic hepatitis in the hepatitis B virus (HBV) and in the HCV groups was not different (25.7 vs. 25.5%). Among those with chronic hepatitis in the HBV group, 4 cases progressed to fulminant hepatic failure, 1 case progressed to the end-stage liver cirrhosis, and 1 case to hepatocellular carcinoma. However, in the HCV group, no case showed progression of chronic hepatitis. The overall mortality in the HBV and HCV groups was 25.3 and 7.8%, respectively (p = 0.001). Among 20 fatal cases in the HBV group 9, cases were liver disease related, but no liver disease related death occurred in the HCV group. In conclusion, HCV as well as HBV infections are quite prevalent and important causes of posttransplant chronic hepatitis, and the clinical course of anti-HCV positive recipients is less aggressive than that of HBsAg-positive recipients. PMID- 7477643 TI - Influence of intrarenal deposits of ciclosporin A on acute renal transplant rejection. AB - Immunohistochemical techniques were used to study the presence of ciclosporin A (CsA) and leukocyte subsets in 36 posttransplant renal biopsy specimens histologically diagnosed as acute graft rejection. Glomeruli from patients with CsA deposits contained more leukocytes (p < 0.05) than glomeruli from tissues without deposits. In contrast, the interstitium from patients without deposits contained significantly more B lymphocytes than interstitia from kidneys with CsA deposits. In both glomeruli and interstitia, the CD4/CD8 ratios were higher in tissues without deposits, although the difference was not significant. The plasma levels of creatinine increased with the intensity of renal CsA deposits, and significantly more patients on hemodialysis had deposits as compared with patients not on hemodialysis. Our findings suggest two types of acute nonvascular rejection: (1) predominantly interstitial, with a good prognosis, characterized by low numbers of intrarenal CsA deposits and a predominance of B lymphocytes and (2) predominantly glomerular, with a poor prognosis, characterized by high levels of intrarenal CsA and a predominance of CD8-positive cells and macrophages. PMID- 7477644 TI - Efficacy and side effects of cyclosporin A in nephrotic syndrome of childhood. AB - The efficacy and side effects of the oral administration of cyclosporin A (Cyc-A) were investigated in 13 children with the nephrotic syndrome. Two of these children were steroid-dependent, 9 were resistant to steroids, and 2 displayed frequent relapses. The diagnosis of 6 patients was mesangioproliferative glomerulonephritis (MezPGN), of another 6 mesangiocapillary glomerulonephritis, and of 1 minimal change disease. Cyc-A was administered for 6 months at a dosage of 7 mg/kg/day with continual monitoring of the serum levels. At the end of the treatment period, 7 of the patients (53.8%) had a total remission, 5 (38.4%) a partial remission, and 1 (7.8%) did not respond to treatment. While total remission was achieved in 4 patients who had responded well to steroids, this was true for only 3 (33.3%) of the 9 patients who had not benefited from steroids. The effects and side effects of Cyc-A on histopathological morphology were also investigated by means of posttreatment biopsies. While histopathological improvement was seen in 3 patients with MezPGN, there was evidence of deterioration in 1 patient with MLH. PMID- 7477645 TI - Renal tubular function modulates urinary levels of interleukin-6. AB - This study was designed to determine whether urinary excretion of IL-6 can be modified as a consequence of deterioration in renal tubular function. In patients with Kawasaki disease, IgA nephropathy and renal hypoplasia, IL-6 and beta 2 microglobulin were measured in the plasma and urine, and the excreted fractions of filtered IL-6 (FEIL-6) and beta 2-microglobulin (FE beta 2-microglobulin were calculated. Patients with renal hypoplasia had elevated FE beta 2-microglobulin which was associated with increased FEIL-6, and there was a significant relationship between these variables as the severity of renal hypoplasia increased. It was concluded that IL-6 in renal tubular fluid may be reabsorbed, catabolized as well as excreted by the renal tubule and that the level of urinary IL-6 excretion could be influenced not only by mesangial proliferation but also renal tubular dysfunction. PMID- 7477646 TI - Creatinine clearance: an inadequate marker of renal filtration in patients with early posthepatitic cirrhosis (Child A) without fluid retention and muscle wasting. AB - Glomerular filtration rate (GFR) was measured in 19 patients with Child A liver cirrhosis by comparing the endogenous creatinine clearance with inulin clearance. Inulin clearance averaged 90 +/- 4.4 ml/min x 1.73 m2, while creatinine clearance averaged 122 +/- 7 ml/min x 1.73 m2 (p < 0.001). The overestimation of GFR by creatinine was present in 18 of 19 patients and was inversely correlated with inulin clearance (r = -0.452, p < 0.04). The data point to the unsuitability of creatinine as a marker of filtration in early posthepatitic cirrhosis (Child A). PMID- 7477648 TI - Hemodialysis for elimination of the nonionic contrast medium iohexol after angiography in patients with impaired renal function. AB - We examined the kinetics of contrast agent elimination during hemodialysis in 7 patients with end-stage renal disease on regular hemodialysis treatment (group I) and in 13 patients with impaired renal function (serum creatinine 214-657 mumol/l; group II). The nonionic agent iohexol was administered at a dose of 0.4 4.5 g/kg, and a 6-hour hemodialysis was performed with 1-18 h delay. This procedure removed 60-90% (mean 77%) of the iohexol present in the circulation at the start of dialysis treatment. The mean extraction ratio across the dialysis membrane was 0.47 and was inversely related to blood flow. The total clearance of iohexol was 70.4 +/- 24.6 ml/min and was very close to dialyzer clearance, as estimated from blood flow and extraction ratio. The plasma iohexol level after dialysis was related to the dose administered, iohexol clearance, and the patients' body weight. During peritoneal dialysis (36-60 liters dialysis fluid), 43-72% of the iohexol dose was removed from the patients' circulation. In patients of group II no further impairment of the renal function (increase of serum creatinine) in conjunction with angiography was observed. We conclude that hemodialysis and peritoneal dialysis are effective methods for removal of iohexol. Our observations suggest also that accelerated elimination of contrast media by prophylactic dialysis can be beneficial in preventing further reduction in renal function after angiographic procedures in high-risk patients. PMID- 7477649 TI - Subclinical signs of kidney dysfunction following short exposure to silica in the absence of silicosis. AB - Signs of kidney dysfunction detectable in urinary protein excretion were searched for in a group of 86 silica-exposed workers who were compared to 86 control subjects matched for age, smoking status and body mass index. No worker had any clinical, spirometric or radiographic sign of silicosis, and exposure duration averaged 15.2 months (range: 11-20). An increase in the urinary excretion of albumin, transferrin, retinol-binding protein and N-acetyl-beta-D-glucosaminidase was found in the exposed group, and the prevalences of pathological values were also elevated in this group. By contrast, both groups had similar serum levels of creatinine and beta 2-microglobulin. These results strongly suggest that occupational exposure to silica may lead to subclinical renal effects after less than 2 years and in the absence of silicosis. PMID- 7477647 TI - Modification of cisplatin-induced renal p-aminohippurate uptake alteration and lipid peroxidation by thiols, Ginkgo biloba extract, deferoxamine and torbafylline. AB - To determine whether inhibition of lipid peroxidation modifies cisplatin-induced changes of renal p-aminohippurate (PAH) uptake, we examined the effects of various radical scavengers and torbafylline on cisplatin-induced lipid peroxidation and PAH accumulation changes in rat renal cortical slices. Renal cortical slices were incubated with different cisplatin concentrations (0.3, 0.6, 1.0 mg/ml) in the presence of either glutathione, N-acetylcysteine, the iron chelator deferoxamine, Ginkgo biloba extract or the xanthine derivate torbafylline. Lipid peroxidation monitored as the production of malondialdehyde (MDA) was stimulated by increasing cisplatin concentrations in a dose-related manner. At a cisplatin concentration of 1.0 mg/ml, MDA production was twofold compared to controls (0.69 +/- 0.06 vs. 1.36 +/- 0.07 nmol/mg; p < 0.05). In turn, cisplatin decreased PAH uptake of kidney slices dose-dependently from 13.3 +/- 1.3 to 2.6 +/- 0.2 (p < 0.01). All agents tested inhibited cisplatin-induced lipid peroxidation; however, at a cisplatin concentration of 1.0 mg/ml, none of them prevented the decline of cisplatin-induced PAH uptake. Of the agents tested, deferoxamine proved to be the most effective antioxidant, completely inhibiting cisplatin-induced lipid peroxidation but in contrast preventing the decrease in PAH uptake only at a cisplatin concentration of 0.3 mg/ml. No strict association between lipid peroxidation and decline of PAH uptake was found, suggesting that lipid peroxidation may only in part participate in cisplatin-induced alterations of PAH uptake. PMID- 7477650 TI - Urinary trehalase activity is a useful marker of renal proximal tubular damage in newborn infants. AB - To clarify the reliability of urinary trehalase activity as a marker of cellular proliferation and/or damage of renal proximal tubules, the activity was examined in healthy newborn infants or infants treated with tobramycin, a drug known as causing tubular cell damage. Eighty-one newborn infants (56 mature infants and 25 premature infants) were enrolled in the study. Urinary trehalase was examined using a spot urine sample during the first 7 days of age and on the 10th day of age. A good positive correlation was observed between urinary trehalase activity/creatinine ratio (T/Cr) on the 10th day of age and conceptional age or body weight (n = 46, r = 0.58, p < 0.001). Urinary trehalase of 29 healthy mature infants was higher during the first few days of age, after which it decreased to an almost steady level. Urinary trehalase of 6 premature infants during the first few days of age was significantly lower than that of mature infants, after which it increased and became equal to that of the mature infants on the 7th day of age. Treatment with ampicillin (100 mg/kg) and tobramycin (5 mg/kg) of 6 mature infants with pneumonia for 6 days resulted in a significant elevation of the urinary T/Cr. The extent of this elevation was greater than that of the urinary N acetyl-beta-D-glucosaminidase (NAG) activity/creatinine ratio (NAG/Cr). A significant correlation was observed between the urinary T/Cr and the urinary NAG/Cr (r = 0.67, p < 0.01) or gamma-glutamyl transpeptidase/creatinine ratio (r = 0.48, p < 0.01).(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7477651 TI - Epstein-Barr virus activity in patients on chronic hemodialysis. AB - Patients with uremia are susceptible to viral infections, especially to Epstein Barr virus (EBV). Sixty-one patients with end-stage renal diseases on chronic hemodialysis (HD), 14 patients with impaired renal function (CRF), and 27 healthy controls were studied with regard to EBV infection. Uremic patients (HD and CRF) had a significantly higher incidence of EBV infection and higher titers of anti EBV VCA-IgG antibody than healthy controls. The anti-EBNA-1 titer was significantly higher in patients whose dialysis period was more than 3 months than in whom the dialysis period was 3 months or less. Immunoblotting analysis also showed stronger EBNA-1 signals in hemodialysis patients than EBNA-2, which was strongly detected in the CRF group and in healthy controls. EBV DNA was detected by Southern blot hybridization after PCR amplification of peripheral leukocytes, and occurred at a greater incidence in hemodialysis patients than in the other groups. Taken together, these results demonstrated that hemodialysis patients had persistent EBV infection. PMID- 7477652 TI - Angiotensin-converting enzyme polymorphism and development of diabetic nephropathy in non-insulin-dependent diabetes mellitus. AB - We determined the distribution frequency of angiotensin-converting enzyme insertion/deletion (I/D) polymorphism in 111 Japanese patients with non-insulin dependent diabetes mellitus (NIDDM) of at least 10 years duration (80 patients with diabetic nephropathy and 31 patients without nephropathy) and 76 healthy Japanese controls. Patients with diabetic nephropathy showed an excess of the ID genotype compared with patients without nephropathy (p < 0.02) and less of the II genotype compared with healthy controls (p < 0.01) and patients without nephropathy (p < 0.01). NIDDM patients with the II genotype have a decreased risk for the development of diabetic nephropathy. PMID- 7477653 TI - Inhibition of puromycin-induced renal injury by a superoxide dismutase derivative with prolonged in vivo half-life. AB - To know the possible involvement of reactive oxygen species and the site(s) of their action in puromycin aminonucleoside (PAN)-induced renal injury, two types of superoxide dismutase (SOD) derivatives were synthesized: one (SM-SOD) circulates bound to albumin with a half-life of 6 h and the other (AH-SOD) linked with hexamethylenediamines rapidly undergoes glomerular filtration and accumulates in renal proximal tubule cells without being excreted in urine. When injected intravenously to the rat, PAN induced a marked proteinuria, increased plasma levels of cholesterol and triglyceride, and suppressed the growth of animals. Intravenously administered SM-SOD significantly inhibited such changes induced by PAN. However, native SOD which rapidly undergoes urinary excretion failed to inhibit the renal injury caused by PAN. Though AH-SOD markedly accumulated in renal proximal tubule cells, it also failed to inhibit the renal injury. These results suggested that superoxide and/or its hazardous metabolite(s) in and around the renal glomerulus, but not in tubule cells, may play critical roles in the pathogenesis of PAN-induced renal injury. PMID- 7477654 TI - Acute and long-term changes observed in imprints of mouse mesothelium exposed to glucose-enriched, lactated, buffered dialysis solutions. AB - Solutions for peritoneal dialysis (PD), the hyperosmolarity of which is obtained with glucose, have been shown to initiate and maintain a situation of continuous mesothelial cell injury associated with a process of regeneration which also takes place continuously. The present study was done using the in vivo and almost in situ technique of mesothelial cell imprints. Acute exposure to 4.25% glucose solution at pH 5.2 and 7.0 induced higher mitotic activity, defective cytokinesis, and reduced cell viability. Long-term exposure (15 and 30 days) to both 4.25% glucose solutions was associated with a reduced population density, increased surface area, and lower mesothelial cell viability, regardless of the pH. The use of 1.5% glucose fluid showed that this effect was dose related. After 30 days of recovery, mesothelial cells exposed to the high-glucose solution at both pH 5.2 and 7.0 appeared repopulated by small cells and showed evidence of defective cytokinesis. So far, it appears that the alterations observed after long-term exposure of the mesothelium to PD fluid are mainly caused by the high concentration of glucose per se. The additional effects of hyperosmolarity are still unclear, whereas the eventual role of low pH, at least in the experimental model used here, can be defined as less than marginal. PMID- 7477655 TI - Effect of colchicine on schistosoma-induced renal amyloidosis in Syrian golden hamsters. AB - Seventy Syrian golden hamsters were infected with Schistosoma haematobium, and 10 uninfected hamsters served as negative controls. Of the schistosome-infected hamsters, 10 served as positive controls (infected but untreated) and the rest (60 hamsters) received treatment. In 30 hamsters treatment was given 9 weeks after infection (before the appearance of renal amyloidosis) and in the other 30 it was given after the appearance of amyloid deposits, 15 weeks after infection. Each treatment group was subdivided into 3 groups (10 hamsters each) in which treatment was either anti-schistosomal alone, combined anti-schistosomal and colchicine, or colchicine alone. Eighteen weeks after infection half of the animals in each group were sacrificed, while the rest were sacrificed 24 weeks after infection. Kidney specimens were evaluated semiquantitatively for renal amyloid deposits. Significant reductions in renal amyloid deposits and proteinuria were observed when combined treatment was given. This was nearly complete with early treatment and only partial when treatment was given late. When colchicine was given alone, a partial but significant reduction in proteinuria with no recognizable effect on renal amyloid deposits was observed. We conclude that colchicine is effective for the prevention and cure of schistosome-related renal amyloidosis in golden hamsters. PMID- 7477656 TI - Decrease of renal endothelin 1 content and gene expression in diabetic rats with moderate hyperglycemia. AB - To investigate the intrarenal endothelin 1 (ET-1) synthesis in streptozocin (STZ) diabetic rats with moderate hyperglycemia, we measured plasma ET-1, renal ET-1 mRNA, and renal tissue ET-1 levels. The renal ET-1 mRNA expression progressively decreased from the 2nd to the 6th week after induction of diabetes by STZ. The renal ET-1 mRNA expression and the renal tissue ET-1 content were significantly reduced in 8 diabetic rats with a mean blood glucose level of 21.0 +/- 0.4 mM as compared with 7 normal rats sacrificed at the 6th week after STZ or citric buffer injection. The reduction of renal ET-1 and mRNA levels was ameliorated in 9 diabetic rats with a mean blood glucose level of 6.9 +/- 0.7 mM after strict glycemic control by insulin treatment. Kidney weight and glomerular filtration rate in moderately hyperglycemic rats were significantly increased as compared with normal rats at the 6th week after STZ injection. The mean plasma ET-1 levels in moderately hyperglycemic diabetic rats were not different from those of the other two groups. This study demonstrates that moderate hyperglycemia in diabetic rats is associated with a reduction in renal ET-1 synthesis. Whether decreased renal ET-1 synthesis is an adaptive phenomenon of a renal hemodynamic change during the early stage of diabetes is worthy of further investigation. PMID- 7477657 TI - Acute renal infarction and heavy marijuana smoking. AB - The first case of acute renal infarction in a patient with heavy marijuana smoking is described. The patient had no concomitant previous medical history. There was no evidence of a primary or secondary hypercoagulable state. Underlying neoplasm was excluded. Recovery was associated with normalization of transaminases and lactic dehydrogenase but with a persistent triangular hypoechogenic area in the right kidney. The combination of marijuana-associated alterations in systemic blood pressure due to peripheral vasodilation, functional anemia and an adrenergic effect might have been the cause of the arteria renalis thrombosis in our patient, but the real underlying pathophysiological mechanism still remains unresolved. PMID- 7477658 TI - HLA class II display by circulating T lymphocytes in nonsteroidal anti inflammatory nephritis induced by drugs. PMID- 7477659 TI - Torsade de pointes and sudden death in a patient with amyloidosis-associated nephropathy. PMID- 7477660 TI - Lipoprotein(a) levels in hemodialysis patients: relation to glucose intolerance and hemodialysis duration. PMID- 7477661 TI - T-cell receptor beta-chain gene polymorphism and the prognosis of IgA nephropathy in Japanese patients. PMID- 7477662 TI - Immunohistochemical localization of laminin in renal lesions of Balkan nephropathy. PMID- 7477663 TI - Peritoneal equilibration test: time on treatment and reference values. PMID- 7477664 TI - Cellulose acetate and cuprophane for hemodialysis: effects on protein catabolic rate. PMID- 7477666 TI - Epidemiology of vascular dementia. AB - Vascular dementia (VaD) is the second commonest dementia after Alzheimer's disease (AD). Epidemiological studies of this condition suffer from many shortcomings related to definition of the disease, diagnostic criteria and assessment of subjects. The prevalence of VaD increases linearly with age and varies greatly from country to country, ranging from 1.2 to 4.2% of people over 65 years old, even after adjustment for age and sex. The incidence of VaD is more homogeneous than prevalence and is estimated at 6-12 cases per 1,000 persons over 70 years per year. The mean duration of the disease is around 5 years and survival is less than for the general population and for AD. The major risk factors for VaD appear to be hypertension, diabetes, heart disease and stroke. Although some of these risk factors are modifiable, there is no study on efficacy of prevention of VaD. PMID- 7477665 TI - Ethnic differences in stroke: black-white differences in the United States population. SECORDS Investigators. Southeastern Consortium on Racial Differences in Stroke. AB - The US Black (African-American) population has a higher stroke incidence and mortality than the US White population. This article reviews the English language literature relating to observed racial and ethnic differences in stroke mortality, incidence, and risk factors. In addition, Black-White differences in stroke subtype, pathophysiology, outcome, recurrence, and treatment are reviewed. The significance of these racial and ethnic differences and directions for future research are explored. PMID- 7477667 TI - Prevalence of spinal cord injury: an international comparison. AB - Information on the prevalence of spinal cord injury is becoming more important as the life expectancy of survivors is increasing, but few prevalence studies have been published. This paper summarises the internationally available findings on prevalence, including the methodologies and data sources used. Prevalence rates ranged from 11 to 112 per 100,000 population. Methodological difficulties in comparing findings over time, country, defined population and data source are discussed, and a more standardised methodology is recommended. PMID- 7477668 TI - A neurochemical approach for studying response to acetylcholine in Alzheimer's disease. AB - The proposal of cholinomimetic treatment as a rational basis for the therapy of Alzheimer's disease has been prematurely dismissed by some workers on the hypothesis of impaired coupling/signal transduction of postsynaptic cholinergic receptors. Disparity of reports studying such impairment may be due to inappropriate extrapolation of experimental systems to the physiological situation, as well as inadequate consideration of disease epiphenoma. In the present study we have used samples with short duration of terminal coma, collected using techniques to minimise postmortem autolysis, and samples obtained during neurosurgery to examine carbachol stimulated hydrolysis of [3H]phosphatidylinositol (PI) as a marker for receptor/signal transduction integrity. The influence of postmortem delay was also studied using another series of samples and a rat model. While a significant correlation of postmortem delay and carbachol stimulated [3H]PI hydrolysis was found, comparison of pooled neurosurgical and postmortem controls with AD samples revealed no significant reduction. Thus this study concurs with a similar one previously reported here, using [3H]phosphatidylinositol 4,5-bisphosphate (1). They provide evidence for competent receptor-signal transduction events in AD, supporting the use of cholinomimetic therapy for disease treatment. PMID- 7477669 TI - Effect of chronic administration of phenytoin on regional monoamine levels in rat brain. AB - Phenytoin (DPH) is a widely used anticonvulsant drug but a conclusive mode of action is not yet clear. This study was undertaken to assess the effects of chronic administration of DPH on monoamine levels. DPH (50 mg/kg body weight) was administered to adult male Wistar rats by intraperitoneal injections for 45 days and the regional brain levels of norepinephrine (NE), dopamine (DA) and serotonin (5-HT) were assayed using high performance liquid chromatographic (HPLC) method. The experimental rats revealed no behavioral deficits of any kind nor body and brain weight deficits were observed. Increased NE levels were observed after DPH administration in motor cortex (P < 0.05), striatum-accumbens (P < 0.01) and hippocampus (P < 0.01), whereas, NE level was decreased in brain stem (P < 0.05). DA levels were increased in striatum-accumbens (P < 0.05), hypothalamus (P < 0.001) and cerebellum (P < 0.001) but decreased in brainstem (P < 0.01). In DPH treated rats, 5-HT levels were increased in motor cortex (P < 0.001) but decreased in cerebellum (P < 0.001) when compared to control group of rats. The present study suggest that chronic administration of DPH induces alterations in monoamine levels in specific brain regions. DPH seems to mediate its anticonvulsant action by selectively altering the monoamine levels in different brain regions. PMID- 7477670 TI - The iron component of sodium nitroprusside blocks NMDA-induced glutamate accumulation and intracellular Ca2+ elevation. AB - These studies were designed to compare the effects of nitric oxide (NO) generating compounds with those of several iron containing compounds which do not generate NO on glutamate receptor function. Stimulation of primary cultures of cerebellar granule cells with N-methyl-D-aspartate (NMDA) or kainate results in the elevation of intracellular calcium ([Ca2+]i) and cGMP and the release of glutamate. The iron containing compounds, sodium nitroprusside (SNP), potassium ferrocyanide (K4Fe(CN)6) and potassium ferricyanide (K3Fe(CN)6) decrease the NMDA induced release of glutamate. SNP is the only compound of the above 3 agents which generates NO. A non-iron, NO generating compound, S-nitroso-N acetylpenicillamin (SNAP), has no effect on the NMDA-induced glutamate release. Potassium ferrocyanide (Fe II), but not potassium ferricyanide (Fe III), blocks NMDA-induced cGMP elevations after 3 min exposure times. This contrasts with the NO generating compounds (both SNP and SNAP) which elevate cGMP levels. Furthermore, both potassium ferrocyanide (Fe II) and SNP (Fe II) suppress the elevation of [Ca2+]i induced by NMDA but neither potassium ferricyanide (Fe III) nor SNAP are effective in this regard. These effects are also independent of cyanide as another Fe II compound, ferrous sulfate (FeSO4) is also able to suppress NMDA-induced elevations of [Ca2+]i. SNP was unable to suppress kainate receptor functions. Collectively, these results indicate that Fe II, independently of NO, has effects on NMDA receptor function. PMID- 7477671 TI - Energy metabolism in hypoxic astrocytes: protective mechanism of fructose-1,6 bisphosphate. AB - The protective effects of fructose-1,6-biphosphate (FBP) during hypoxia/ischemia are thought to result from uptake and utilization of FBP as a substrate for glycolysis or from stimulation of glucose metabolism. To test these hypotheses, we measured CO2 and lactate production from [6-14C]glucose, [1-14C]glucose, and [U-14C]FBP in normoxic and hypoxic cultured astrocytes with and without FBP present. FBP had little effect on CO2 production by glycolysis, but increased CO2 production by the pentose phosphate pathway. Labeled FBP produced very small amounts of CO2. Lactate production from [1-, and 6-14C]glucose increased similarly during hypoxic hypoxia; the increase was independent of added FBP. Labeled lactate from [U-14C]FBP was minimal. We conclude that exogenous FBP is not used by astrocytes as a substrate for glycolysis and that FBP alters glucose metabolism. PMID- 7477672 TI - Changes in organic solutes, volume, energy state, and metabolism associated with osmotic stress in a glial cell line: a multinuclear NMR study. AB - Diffusion-weighted in vivo 1H-NMR spectroscopy of F98 glioma cells embedded in basement membrane gel threads showed that the initial cell swelling to about 180% of the original volume induced under hypotonic stress was followed by a regulatory volume decrease to nearly 100% of the control volume in Dulbecco's modified Eagle's medium (DMEM) but only to 130% in Krebs-Henseleit buffer (KHB, containing only glucose as a substrate) after 7 h. The initial cell shrinkage to approx. 70% induced by the hypertonic stress was compensated by a regulatory volume increase which after 7 h reached almost 100% of the control value in KHB and 75% in DMEM. 1H-, 13C- and 31P-NMR spectroscopy of perchloric acid extracts showed that these volume regulatory processes were accompanied by pronounced changes in the content of organic osmolytes. Adaptation of intra- to extracellular osmolarity was preferentially mediated by a decrease in the cytosolic taurine level under hypotonic stress and by an intracellular accumulation of amino acids under hypertonic stress. If these solutes were not available in sufficient quantities (as in KHB), the osmolarity of the cytosol was increasingly modified by biosynthesis of products and intermediates of essential metabolic pathways, such as alanine, glutamate and glycerophosphocholine in addition to ethanolamine. The cellular nucleoside triphosphate level measured by in vivo 31P-NMR spectroscopy indicated that the energy state of the cells was more easily sustained under hypotonic than hypertonic conditions. PMID- 7477673 TI - Chick embryo retina development in vitro: the effect of insulin. AB - In this paper we study the development of chick embryo retina cultured in vitro and the effects exerted by insulin. Retinas were removed from 7-day embryos and cultured in serum- and hormone-free medium for 7 additional days. Under these conditions retinal cells survived and underwent cholinergic differentiation, as previously ascertained by Hausman et al. (Dev. Brain Res., 1991, 59: 31-37). However, a great retardation of development was noted compared to uncultured control, 14-day retina. In fact both wet weight and DNA and protein content increased much slower than in ovo and the tubulin content decreased below even the starting value. In addition, although after 7 days in culture retinal cells were organized in identifiable layers, nevertheless the typical organization equivalent to 14-day in ovo retina was absent. The addition of insulin in the medium markedly increased the wet weight of cultured retinas, their protein content and the level of tubulin pools, particularly that of non-assembled fraction. Nevertheless insulin did not modify DNA synthesis and did not induce the increment of both neuron specific enolase and actin. Morphological observations show that insulin markedly increased the number and the thickening of the fiber layers. These results, together with the facts that retina synthesizes and secretes insulin and possesses specific insulin receptors suggest that insulin can have autocrine or paracrine regulatory functions in retinal development by exerting a general effect on retinal growth and a more specific one on tubulin production. PMID- 7477675 TI - Endogenous serotonin release from the dopamine-deficient striatum of the weaver mutant mouse. AB - In addition to an altered dopaminergic input, the striatum of the weaver mutant mouse (wv/wv) has increased serotonin tissue content and uptake compared to the wild-type mouse (+/+). To gain information regarding the functional status of serotonergic inputs to the wv/wv striatum, endogenous serotonin release from wv/wv and +/+ striatum was measured under basal conditions as well as in the presence of fenfluramine or elevated concentrations of potassium (K+). Fractional basal release of serotonin from the +/+ striatum was significantly greater than that from the wv/wv striatum. In the presence of K+, evoked release (stimulated release minus basal release) was greater from the +/+ striatum than from the wv/wv striatum. In the presence of fenfluramine, evoked serotonin release was greater from the wv/wv striatum compared to the +/+ striatum. These data are consistent with the involvement of an additional transmitter(s) in modulating serotonin release to a greater extent in the wv/wv than the +/+ striatum. The data on fenfluramine-stimulated serotonin release suggest that the additional serotonin content found in the wv/wv striatum is in a releasable pool but that striatal serotonin release might be attenuated more in wv/wv than in +/+ mice. PMID- 7477674 TI - Change in fluidity of brain endoplasmic reticulum membranes by oxygen free radicals: a protective effect of stobadine, alpha-tocopherol acetate, and butylated hydroxytoluene. AB - Effect of various oxygen free radical generating systems and an oxidant H2O2 on brain endoplasmic reticulum (ER) membrane fluidity was examined using fluorescent membrane probe 1,6-diphenyl-1,3,5-hexatriene, DPH. The relative potency of free radical generating systems to decrease membrane fluidity increased in this order: FeCl3-EDTA, FeSO4-EDTA, FeSO4-EDTA/hydrogen peroxide. Potency to decrease membrane fluidity correlated well with these systems' potencies to induce lipid peroxidation, as detected by conjugated diene formation. Treatment of ER membranes with H2O2 had no effect on fluidity or conjugated diene formation. Using the two most potent free radical generating systems, FeSO4-EDTA and FeSO4 EDTA/hydrogen peroxide, a protective effect of the novel antihypoxic and antiarrhytmic drug stobadine was tested. Stobadine and two well-known antioxidants, alpha-tocopherol acetate and butylated hydroxytoluene, demonstrated the ability to prevent free radical induced alterations in ER membrane fluidity. These results provide new evidence of stobadine's protective effect on membranes attacked by oxygen free radicals. PMID- 7477676 TI - Alteration of opioid peptide concentrations in the rat pituitary following survivable closed head injury. AB - Concentration changes of methionine enkephalin-like immunoreactivity (ME-li) and beta-endorphin-like immunoreactivity (BE-li) in the rat pituitary following diffuse brain injury were studied. Closed head injury was induced by a weight drop trauma device (450 g x 2 m). The level of closed head injury used in this study altered the pituitary opioid peptide concentrations. The level of ME-li did not change in the experimental groups 3 hours, 10 hours, 24 hours, and 3 days after the trauma, but significantly increased by 34% 10 days after the trauma. BE li remained constant 3 hours and 10 hours following the injury, increased by 48% at 24 hours, and remained at this level for 10 days after the trauma (44% at 3 days, and 40% at 10 days). The levels of ME-li and BE-li in the control sham operated rats did not change during these times. The present measurements of BE li and ME-li in the pituitary indicate that the opioid peptides that derive from two different neuropeptidergic systems, proopiomelanocortin (POMC) and preproenkephalin A, respectively, may participate in the pathophysiology of a closed head injury. PMID- 7477677 TI - Expression of low-molecular-weight neurofilament (NF-L) mRNA during postnatal development of the mouse brain. AB - A regional Northern blot analysis demonstrated that the highest levels of NF-L mRNA in the adult mouse brain are present in brain stem followed by mid-brain, with lower levels found in neocortex, cerebellum, and hippocampus. The study was extended to the cellular level over course of postnatal development using in situ hybridization. This developmental analysis revealed that the expression of NF-L mRNA closely follows the differentiation pattern of many large neurons during postnatal neurogenesis. Neurons which differentiate early such as Purkinje, mitral, pyramidal, and large neurons of brain stem and thalamic nuclei, expressed high levels of NF-L mRNA at postnatal day 1. Early expression of NF-L mRNA may be required for the maintenance of the extensive neurofilament protein networks that are detected within the axons of larger neurons. Smaller neurons which differentiate later, such as dentate gyrus granule cells, small pyramidal and granule cells of the neocortex, and granule cells of the cerebellum, exhibit a delayed expression of NF-L mRNA. PMID- 7477680 TI - Acidosis-induced modifications of high-affinity choline uptake by synaptosomes: effects of pH readjustment. AB - Acidosis (pH 6.0) led to significant decrease in high-affinity choline uptake by rat brain synaptosomes. The effects persisted following pH readjustment (7.4) of the incubation medium, consisting of decrease in both Km and Vmax of the affinity system. pH readjustment coincided with synaptosomal leakage of lactate dehydrogenase (LDH) and with instability of the synaptosomal suspension as evidenced from turbidity modifications of the preparation. LDH leakage occurred when acidosis was performed with lactic acid, whereas it was not seen following H3PO4 acidosis, probably because of the rapid diffusion of the protonated from of lactic acid across membranes. Turbidity modifications of the suspension were prevented by EDTA. The present results indicate that acidosis to pH level comparable to what is observed in brain ischemia is deleterious for cholinergic mechanisms. They also suggest that alkaline pH shifts that occur after blood reperfusion of ischemic brain tissue might be critical for the survival of cells. PMID- 7477679 TI - Glutamine synthetase (GS) expression is reduced in senile dementia of the Alzheimer type. AB - Glutamine synthetase (GS), a metabolic marker of the mature astrocyte, was investigated in the temporal neocortex of postmortem brain samples of 8 cases, either not demented or affected by senile dementia of the Alzheimer type. A negative correlation between the GS protein level and the density of both classical beta A4 deposits and senile plaques was evidenced. Such a correlation for GS underlies a dysfunction of the astroglial metabolism and particularly of the glutamate and ammonia neutralization. Since GS is sensitive to oxidative lesioning, the changes in GS level that were observed, occurring at the posttranslational stage, might reflect oxidative damage and have severe consequences on the pathological cascade of events. PMID- 7477678 TI - Endogenous excitatory amino acid release from brain slices and astrocyte cultures evoked by trimethyltin and other neurotoxic agents. AB - Trimethyltin (TMT) is a toxic alkyltin compound that is known to produce neuronal necrosis in the CNS. The present study examined the effects of TMT on the release of excitatory amino acids (EAA) from cortical slices prepared from adult and aged (24 months old) rats. The calcium dependence of TMT-induced EAA efflux was evaluated and compared to other neurotoxic agents. The actions of TMT were also evaluated in an astrocyte culture model to assess glial contributions to TMT induced EAA efflux. TMT (10-1000 microM) evoked a dose-related increase in GLU and ASP efflux during a 30 min incubation period and this efflux was sustained or slightly higher during a 15 min recovery period. TMT-stimulated GLU efflux was not altered in aged rats. TMT-induced GLU efflux was significantly reduced by removing extracellular calcium and including 10 microM EGTA in the incubation media. Calcium channel blockers (nifedipine, verapamil, flunarizine, amiloride, neomycin) and MK-801 did not significantly attenuate TMT-induced GLU efflux. Diltiazem (25 microM) produced modest but inconsistent reductions in TMT-induced GLU efflux from brain slices, and significantly inhibited the leakage of lactate dehydrogenase (LDH) from TMT-treated astrocyte cultures. TMT did not increase GLU efflux from glial cultures during a 30 min incubation period, but did significantly elevate GLU efflux during the 15 min recovery period. TMT evoked the release of EAA by both calcium dependent and independent mechanisms in brain slices. TMT at high concentrations also produced a delayed increase in glial GLU efflux. These studies suggest that excitotoxic mechanisms may contribute to TMT induced neurotoxicity. PMID- 7477681 TI - Down-regulation of blood-brain glucose transport in the hyperglycemic nonobese diabetic mouse. AB - The intracarotid injection method has been utilized to examine blood-brain barrier (BBB) glucose transport in hyperglycemic (4-6 days) mice. In anesthetized mice, Brain Uptake Indices were measured over a range of glucose concentrations from 0.010-50 mmol/l; glucose uptake was found to be saturable and kinetically characterized. The maximal velocity (Vmax) for glucose transport was 989 +/- 214 nmol.min-1.g-1. and the half-saturation constant estimated to be 5.80 +/- 1.38 mmol/l. The unsaturated Permeability Surface area product (PS) is = 171 + 8 microliters.min.-1.g-1. A rabbit polyclonal antiserum to a synthetic peptide encoding the 13 C-terminal amino acids of the human erythrocyte glucose transporter immunocytochemically confirmed the presence of the GLUT1 isoform in non-obese diabetic (NOD) mouse brain capillary endothelia. These studies indicate that a down-regulation of BBB glucose transport occurs in these spontaneously hyperglycemic mice; both BBB glucose permeability (as indicated by PS product) and transporter maximal velocity are reduced (in comparison to normoglycemic CD-1 mice), but the half-saturation constant remains unchanged. PMID- 7477682 TI - The effect of ethanol on HSP70 in cultured rat glial cells and in brain areas of rat pups exposed to ethanol in utero. AB - Prenatal exposure to alcohol is associated with a cluster of symptoms called Fetal Alcohol Syndrome with a characteristic pattern of neuroanatomy and biochemical changes. In recent years it has been shown, that stress exposed cells rapidly increase transcription and translation of heat shock protein genes resulting in an increased appearance of these proteins. It has also been found that heat shock proteins, especially the HSP70 family play a role as molecular chaperons maintaining the native conformation of proteins and participating in protein transport in particular cellular compartments. The aim of this study was to determine the effect of chronic maternal alcohol consumption on HSP70 content in the different regions of the brain of the newborn rats as well as to examine in vitro the effect of ethanol on HSP70 content in cultured glial cells. Chronic maternal ethanol consumption resulted in increased HSP70 in the following regions of developing brain: hippocampus, cerebellum, olfactory bulbs, frontal cortex and septum. Moreover, ethanol applied in vitro, increased HSP70 content in primary astroglial cultures and astrocytes but not in oligodendrocyte cultures. The above described changes may be important in brain maturation and may play a role in Fetal Alcohol Syndrome. PMID- 7477684 TI - Secretion of transforming growth factor-beta 1 and -beta 2 by malignant glioma cells. AB - The secretion of transforming growth factor-beta (TGF-beta), a growth inhibitory factor with immunosuppressive properties, was investigated in one glioblastoma cell line and seven surgically resected malignant glioma cells. Cultured cells from surgically resected tumors were examined immunohistochemically for glial fibrillary acidic protein (GFAP) and S-100 protein. The levels of TGF-beta 1 and TGF-beta 2 in culture supernatants from malignant glioma cells were determined by a specific bioassay using anti-TGF-beta 1 and anti-TGF-beta 2 antibodies. Two glioblastoma cell lines were cultured in the presence of TGF-beta 1 or TGF-beta 2 to assess the effect of TGF-beta on the growth of glioblastoma cells. Cultured cells from surgically resected tumors were positive for both GFAP and S-100 protein. Both active and latent forms of TGF-beta 1 and TGF-beta 2 were detected in the culture supernatants from malignant gliomas, except in one patient with anaplastic astrocytoma which secreted only latent forms of TGF-beta 1 and TGF beta 2. There was no statistical difference in the levels of TGF-beta 1 and TGF beta 2 in glioblastomas and anaplastic astrocytomas. Neither TGF-beta 1 nor TGF beta 2 affected the growth of glioblastoma cells. These findings suggest that most malignant glioma cells secrete both TGF-beta 1 and TGF-beta 2, can convert TGF-beta from a latent to active form, and may suppress cytokine secretion by activated lymphocytes in vivo as well as in vitro. PMID- 7477683 TI - Biochemical and pharmacological studies on a lethal neurotoxic polypeptide from Phoneutria nigriventer spider venom. AB - Fractionation of Phoneutria nigriventer spider venom by gel filtration and HPLC yielded a few fractions that induced different effects when administered intraperitoneally in mice. One of these fractions, PF3, was chemically characterized as a cysteine-rich polypeptide of approximately 8360 MW. Administered at 0.1 mg/kg, i.p., PF3 induced a progressive paralysis and death of mice within 30 minutes. Partial sequence analysis of PF3 revealed certain homologies with other spider toxins already described, particularly omega-AGAIIA (60%) from Agelenopsis aperta. Pharmacological characterization carried out in superfused chopped rat striatal tissues preloaded with [3H]-Dopamine ([3H]-DA) showed that PF3 (0.1 microgram/ml) decreased the [3H]-DA release induced by 20 mM K+ or 100 microM glutamate without changing the basal release. At 1 microgram/ml, PF3 inhibited 33% of the basal release of [3H]-DA; the transmitter release stimulated by K+ or by glutamate was reduced by respectively, 87% and 77% of corresponding control values. PF3 (0.1 micrograms/ml) altered the dose-response curves of glutamate (1 microM-10 mM), by reducing by 36% of its maximal effect. Naloxone (1 microM) did not influence the effect of PF3. The results indicate that PF3 inhibits the [3H]-DA release induced by membrane depolarization or that mediated by NMDA glutamate receptors. These data suggest that the mechanism of action of PF3 may involve a blockade of Ca2+ channels as well as a direct effect on the exocytotic machinery. PMID- 7477686 TI - Correlation between magnetic resonance images and histology in meningiomas: T2 weighted images indicate collagen contents in tissues. AB - The magnetic resonance imaging appearance of 40 surgically confirmed intracranial meningiomas was reviewed to correlate signal intensity on T2-weighted images with histological subtypes of meningioma. A moderate variation of signal intensity was found within a given histological subtype. The mean signal intensity scores on T2 weighted images of the fibrous type of meningioma were significantly lower than those of the other types, and were correlated with the collagen content. The signal intensity of meningiomas on T2-weighted images crudely predicts the histological type. Tumors significantly hypointense compared to the cortex are composed primarily of fibrous elements. PMID- 7477687 TI - Magnetic resonance imaging characteristics of choroid plexus papilloma in the fourth ventricle. AB - The computed tomography (CT) and magnetic resonance (MR) imaging characteristics of two cases of small choroid plexus papilloma within the fourth ventricle are reported. CT showed the tumors as high density areas with postcontrast enhancement. MR imaging showed the tumors as basically isointense areas with marked enhancement by gadolinium-diethylenetriaminepenta-acetic acid. Important MR imaging findings included clear evidence of tumor attachment to the normal choroid plexus, location in the lower part of the fourth ventricle, and separation from the fourth ventricular floor. PMID- 7477688 TI - Hemodynamic evaluation of spinal arteriovenous malformations before and after embolization--preliminary report. AB - This preliminary study investigated local hemodynamic changes in intramedullary spinal arteriovenous malformation (AVM) before and after embolization. 99mTcO4- was injected into the anterior spinal artery feeding the AVM via a Tracker-10 or MAGIC microcatheter. Time-dependent radioisotope images were sequentially obtained in the anteroposterior plane every 0.2 sec before and just after embolization. Local mean transit time (MTT) was then calculated for both the nidus and draining vein and compared before and after embolization. Prior to embolization, MTTs in the nidus and in the draining vein were 1.84 +/- 0.62 (mean +/- SD) and 2.80 +/- 0.69 sec for the five patients, respectively. MTTs in both the nidus and the draining vein were significantly prolonged after embolization to 3.32 +/- 1.14 and 4.90 +/- 0.93 sec, respectively (p < 0.02 and p < 0.005, respectively). In vivo measurements of local hemodynamic changes in the spinal cord during the treatment of spinal AVMs could be achieved. This method may allow investigation of the hemodynamic mechanisms which induce ischemic symptoms in patients with spinal AVM. PMID- 7477689 TI - Ruptured dissecting aneurysm of the peripheral anterior cerebral artery--case report. AB - A 27-year-old male presented with intracranial hemorrhage due to rupture of an idiopathic dissecting aneurysm in the A4 segment of the left anterior cerebral artery (ACA). This is a very rare location. He was successfully treated by resection of the aneurysm without neurological deficits. Surgical intervention is recommended for patients with intracranial hemorrhage due to ruptured dissecting aneurysm of the ACA to prevent rebleeding. PMID- 7477685 TI - Accumulation of nerve growth factor in cerebrospinal fluid and biological activity following neurosurgery. AB - Perioperative nerve growth factor (NGF) levels in cerebrospinal fluid (CSF) of patients with acoustic neurinoma (14 cases), tentorial meningioma (1 case), or subarachnoid hemorrhage (1 case) were examined. Preoperative NGF levels in CSF were below the level of detection in all patients. However, NGF was found to accumulate transiently in CSF following neurosurgery. Pre- and postoperative CSF obtained from a patient with acoustic neurinoma enhanced the proliferation of astrocytes in neuronal cell cultures derived from embryonic rat cortex grown in serum-free defined medium, and increased choline acetyltransferase activity of cholinergic neurons derived from embryonic rat septal area and brainstem. The effect of postoperative CSF on septal and brainstem neurons was more potent than that of preoperative CSF. These results indicate that NGF and non-NGF-type neurotrophic activities accumulate in the CSF following neurosurgery. These neurotrophic activities are probably important in the regeneration of damaged neural networks in the central nervous system. PMID- 7477690 TI - Subarachnoid hemorrhage of unknown origin associated with Weber-Christian disease -case report. AB - A 58-year-old female, who had suffered from Weber-Christian disease for 26 years, presented with subarachnoid hemorrhage. Cerebral angiography showed dilatation of the basilar tip. An operation was performed in the chronic stage. The wall of dilated basilar artery was tough, but that of the right superior cerebellar artery was very thin and three small aneurysms were found on the right middle cerebral artery. Blood levels of fibrin degradation products, plasmin-alpha 2 plasmin inhibitor complex, and thrombin-antithrombin III complex were increased. The abnormality of the coagulation-fibrinolysis system and the fragility of the cerebral arteries related to Weber-Christian disease were probably the cause of the subarachnoid hemorrhage. PMID- 7477691 TI - Cerebellar hemangioblastoma with intracystic hemorrhage--case report. AB - A 45-year-old male presented with sudden onset of severe headache, mild disorientation, and gait disturbance due to intracystic hemorrhage from a cerebellar hemangioblastoma. He was successfully treated with ventricular drainage followed by total removal of the tumor. Reported cases of intracranial hemangioblastoma with massive hemorrhage show male predominance, tend to be solid rather than cystic, and occur in supratentorial more than infratentorial locations. Thin-walled and dilated vessels in the present and other cases were a possible cause of hemorrhage. Massive hemorrhage from hemangioblastoma is rare but needs prompt treatment because of the relatively high mortality. PMID- 7477692 TI - Cerebral gumma mimicking glioblastoma on magnetic resonance images--case report. AB - A 39-year-old female presented with a syphilitic cerebral gumma mimicking small glioblastoma multiforme manifesting as visual impairment. Magnetic resonance imaging showed a low signal intensity area on T1-weighted images and marked small ring-like enhancement less than 1 cm in diameter following the administration of gadolinium. The symptoms and radiological abnormalities improved several weeks after penicillin therapy. The small ring-like enhancement was probably caused by inflammation associated with treponemal infection. A mass with a small ring-like enhancement less than 1 cm in diameter is a useful finding indicating an inflammatory lesion rather than a glial tumor. PMID- 7477693 TI - Diagnosis of brainstem abscess in the cerebritis stage by magnetic resonance imaging--case report. AB - A 52-year-old male presented with a brainstem abscess manifesting as high fever, diplopia, and left hemiparesis. Magnetic resonance (MR) imaging with gadolinium diethylenetriamine pentaacetic acid showed the lesion as a ring-like enhanced mass consisting of a necrotic center with surrounding edema, whereas postcontrast computed tomography revealed no such confirmatory findings. He was treated with antibiotics as the lesion had been detected in the acute cerebritis stage. Serial MR images showed that the lesion decreased remarkably in size. MR imaging can detect brain abscess in the earliest inflammatory stage. PMID- 7477694 TI - [Pineal gland and melatonin]. PMID- 7477695 TI - [Superficial temporal artery to superior cerebellar artery anastomosis via the presigmoid retrolabyrinthine transtentorial approach]. PMID- 7477696 TI - [Characteristic neurological signs in patients with cervical disc disease]. AB - In cervical radiculopathy, neurological findings for the upper extremities and the responsible level of the radices have been precisely detailed. However, in cases of cervical spondylotic myelopathy, it is not always easy to determine the responsible level of the cervical spine solely based on neurological findings. To disclose characteristic neurological lesions of the upper extremities, if any, for different locations of disc disease, 49 patients with cervical disc protrusion at single levels, confirmed by magnetic resonance imaging, were analyzed. Thirty-six patients demonstrated long tract signs (LTS) concomitant with segmental signs in the upper extremities, and 13 patients had no LTS. Except for deep tendon reflexes and the extent of dysesthesia in the upper extremities, there were no characteristic neurological findings, such as muscle atrophy, loss of muscle strength, extent of hypalgesia, or hypesthesia, pointing to the responsible compression site. In the 13 patients without LTS, the extent of dysesthesia and diminution of deep tendon reflexes proved to be completely in agreement with results for radiculopathy: 9 had protruded discs at the C56 level and described dysesthesia in the thumb and the index finger with deep tendon reflexes in the biceps and brachioradialis being diminished. The remaining 4 had protruded discs at the C67 level and described dysesthesia in the middle finger. Their deep tendon reflexes in the triceps were diminished. A clear contrast was observed for the patients with LTS. Four out of 36 patients had C34 protruded discs and did not complain of dysesthesia in any digits. Their deep tendon reflexes were exaggerated in all muscles in the upper extremities.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7477697 TI - [Endovascular treatment of cerebral vasospasm with intra-arterial papaverine infusion]. AB - Thirty-one cases of cerebral vasospasm following subarachnoid hemorrhage were treated with intraarterial papaverine infusion. Symptomatic cases were nineteen, and asymptomatic cases were twelve. Papaverine (120 mg/saline 50 ml, 30 min) was injected superselectively to vasospastic vessels through a microcatheter. The rate of symptomatically improved cases was 63% initially, but about two thirds of those cases had recurrence within a day. The 63% of symptomatic cases showed infarction in spite of papaverine infusion. Three cases of recurrent vasospasm after intra-arterial papaverine underwent PTA and showed good dilatation of vasospastic vessels. The complications of our intra-arterial papaverine were hypotension in two cases, convulsion in one case and transient disturbed consciousness in one case. We experienced no fatal complications. Overall outcome was ADL1 (19%), ADL2 (25%), ADL3 (44%), ADL4 (0%), ADL5 (6%), and death (6%). Since the effect of intra-arterial papaverine infusion is of short duration and weak, combination of PTA and papaverine may be necessary. It is recommended to use papaverine for vasospasm in distal arteries such as M2, A1, A2, and to carry out PTA for proximal arteries such as ICA and M1. PMID- 7477698 TI - [Operative results in fourteen cases of paranasal sinus and anterior cranial fossa lesions surgically treated by an extended transbasal approach]. AB - We describe the outcomes and complications of 14 patients with paranasal sinus and anterior cranial fossa lesions surgically treated by an extended transbasal approach, originally described by Kawakami, in our department. They were 10 patients with malignant tumors, 2 with benign tumors, and 2 with inflammatory diseases. A bifrontal craniotomy was performed using a high coronal skin incision, and the orbital rim and roof were removed after the dissection of the dura mater from the anterior skull base. Transcranial resection of the tumor was performed, and assisted by transnasal and transmaxillar resection using a nasal endoscope. Reconstruction of the anterior skull base was performed with the fasica lata and galeopericranial flap in all cases, temporal musculo-pericranial flap in 3 and free bone flap from the cranial convex in 3. Among 10 patients with malignant tumors (malignant melanoma; 4, squamous cell carcinoma; 2 adenocarcinoma; 1, malignant plemorphic adenoma; 1, chondrosarcoma; 1, and neuroblastoma; 1), total removal was performed in 5 patients and subtotal removal in 5 patients. Though local recurrence of the tumor was recognized in 6 patients, only one underwent additional surgery. Eight patients survived, and 2 patients died of systemic metastasis of the tumor and complications due to liquorrhea. Seven patients obtained a good quality of life, and the mean survival period in 8 patients still living was 27 months after the first surgery. In 2 patients with benign tumors (chordoma and osteoma), partial and total removal was performed. The patient with chordoma was operated on several times by this approach and the transoral approach, respectively. Each had a good postoperative course.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7477699 TI - [Image guided stereotactic biopsy for brain tumors: experience of 71 cases]. AB - Despite advances in the neuroimaging of the brain, an accurate diagnosis of intrinsic lesion of the brain requires tissue sampling and histological verification. Seventy-one patients with intraparenchymal lesion of the brain underwent CT or MRI-directed stereotactic biopsy at Okayama University Hospital between June, 1987 and March, 1995. There were 32 men and 39 women whose ages ranged between 7 and 78 years (mean 46.1 years). All patients underwent preoperative cerebral angiography, high resolution contrast enhanced CT and MRI. The lesions were located in the hemisphere in 40 cases, the thalamus or basal ganglia in 14, the midline (corpus callosum or ventricle) in 11, the pineal region in 4, the suprasellar in one and multiple sites in one. A Brown-Roberts Wells (BRW) CT-directed stereotactic system was used for biopsy under CT guidance. For MRI-directed biopsies a prototype modification of the BRW frame was employed. Target localization was performed using either CT or MRI. Usually one or two targets within the lesion were chosen and target coordinates were calculated using the CT or MRI scan soft ware. Positive diagnosis was obtained in 67 cases and the accuracy of the histological diagnosis was 94.4%. There were 53 gliomas, 4 metastasis, 5 germinomas, 3 malignant lymphomas, one pineoblastoma, one infarction and 4 negative biopsies. Bleeding as a complication due to stereotactic intervention occurred in one patient (1.4%). To patients with potentially inoperable lesions or lesions which might be best treated by chemotherapy or irradiation, modern techniques of neurosurgery now offer the option of precise stereotactic biopsy through small twist-drill burr holes as opposed to open biopsy.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7477700 TI - [A case report of epidermoid carcinoma in the cerebello-pontine angle]. AB - A case of epidermoid carcinoma in the cerebello-pontine (CP) angle is presented. A 42-year-old male was admitted with a complaint of experiencing double vision for four months in January, 1992. During neurological examination, right abducens palsy, right facial dysesthesia, and atrophy of the right temporal muscle were noted. Magnetic resonance (MR) imaging revealed a mass of low intensity in the right CP angle, which was prominently enhanced with gadolinium. Malignancy was suspected because the tumor on MR enlarged rapidly in a month, so the first surgical resection was performed. Suboccipital exploration of the right CP angle was performed in February. At first, a fragile, pearly part of the mass typical of epidermoid was exposed behind the seventh and eighth cranial nerve complex. Then, a grayish, fibrous part was exposed, which involved the fifth cranial nerve and was attached to the tentorium and the brainstem. Histological diagnosis of the fragile part of the tumor revealed a typical epidermoid cyst and that of the fibrous part was squamous cell carcinoma. During postoperative examinations on other parts of the body, such as endoscopic studies of the trachea and the esophagus, no abnormality was shown. Therefore the tumor was diagnosed as a primary intracranial epidermoid carcinoma. Post-operatively, conventional fractionated external-beam focal irradiation was carried out, which caused regression of the residual tumor for eleven months. Subsequently, palsy of the right side of the tongue and paresis of the contralateral side of the extremities and face developed with increase of the right abducens palsy. MR imaging indicated regrowth of the tumor. The second operation via the subtemporal approach was unsuccessful, because the tumor was fibrous and firmly attached to the brainstem.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7477701 TI - [Pineocytoma associated with a cyst: report of a case]. AB - We report a case of pineocytoma associated with an unusually large cyst. A 15 year-old boy was admitted with a complaint of intermittent headache. Neurological examination was normal except for bilateral papilledema. CT scan showed triventricular hydrocephalus and a large mass. A solid portion of the tumor in the pineal and retrothalamic areas was isodense, and contained massive calcification on CT. A huge cystic portion extended far into the infratentorial, supracellebellar area. On MR, the solid tumor showed isointensity on T1-WI and iso- to high intensity on T2-WI, and was enhanced by Gd-DTPA. After a ventriculo peritoneal shunt, the tumor was partially removed through an occipital supratentorial approach. Histological examination revealed a pineocytoma with neuronal and astrocytic differentiations. Reviewing the literature, it is most unusual to find a pineocytoma associated with a huge cyst as was seen in this case. PMID- 7477702 TI - [Frontal acute extradural hematoma due to contrecoup injury: a case report]. AB - Acute extradural hematoma, a collection of blood between the inner table of the skull and the dura, generally occurs in coup craniocerebral trauma with skull fracture. We report here a case of a 52-year-old female who had had a traffic accident. She suffered a serious wound consisting of a subcutaneous hematoma with skull fracture and so called Battle's sign on the left occipital region but not wounded in any other region. CT scan obtained shortly after the injury demonstrated a thin acute extradural hematoma with intracranial air under the subcutaneous hematoma in the left occipital region, and another hematoma with subarachnoid hemorrhage due to contrecoup head trauma in the right frontal region. We treated her conservatively with a common drip. She sometimes vomited during the several hours after admission. CT scan six hours later showed that the contrecoup extradural hematoma had enlarged. Immediately, we carried out the evacuation of the hematoma with decompressive craniectomy. Her scalp, cranium and frontal skull base over the extradural hematoma were without injury, and multiple small haemorrhages were found to have occurred on the surface of the dura that had been separated from the inner table of the skull. After the operation, the patient recovered consciousness. Contrecoup acute extradural hematoma is very rare. It seems that the appearance of hematoma in our case resulted from the frontal dural separation due to distortion of the cranium brought on by the force of the impact and the subsequent gradual growth of the hematoma under the stimulation of several bouts of vomiting. PMID- 7477703 TI - [A case of pineoblastoma successfully treated with surgery, combined chemotherapy of cisplatin and etoposide, and radiotherapy]. AB - A 5-year-old girl was admitted to another clinic because of vomiting and convulsions. She was brought to our clinic after a ventriculoperitoneal shunt was inserted. CT scan on admission in our clinic showed a tumor in the pineal region with tumoral hemorrhage. Tumor markers such as HCG, AFP, CEA, P-LAP were within normal range. A biopsy of the tumor was performed and the histological diagnosis was pineoblastoma. Her recovery was excellent and disseminated metastasis was not recognized. A subtotal removal of the tumor was performed through the occipital transtentorial approach. She had no neurological deficits after surgery. She then received two 5-day cycles of chemotherapy, consisting of intravenous administration of 20 mg/m2/day cisplatin and 60 mg/m2/day etoposide, and craniospinal radiotherapy. After these therapies, the tumor responded and disappeared completely. Follow-up radiographic investigations also demonstrated no abnormal evidence except for brain atrophy. She is attending a primary school without any problems. Pineoblastoma is quite rare and remarkably malignant. Hence, aggressive therapies including surgery, radiotherapy and chemotherapy is indicated for this tumor. PMID- 7477704 TI - [Evaluation by proton magnetic resonance spectroscopy of carotid-cavernous fistula with cortical venous drainage]. AB - Cases with carotid-cavernous fistula (CCF) associated with cortical venous drainage through the sylvian veins are rather rare. However, such cases involve risk for subarachnoid hemorrhage, subcortical hemorrhage and venous infarction due to venous hypertension in the brain. Even without these symptoms, CCF under these conditions provokes disturbance in cerebral metabolism. We report two cases of CCF associated with cortical venous drainage evaluated by proton magnetic resonance spectroscopy (1H-MRS). Case 1: A 56 year-old female suffered from a right CCF associated with cortical venous drainage through the sylvian veins after trauma. Before embolization with a detachable balloon catheter, the ratios of N-acetyl-aspartate (NAA)/Choline (Cho) and NAA/Creatine (Cr) in 1H-MRS on the right temporo-basal ganglia were lower than those in normal volunteers. After curative balloon embolization of the CCF, serial 1H-MRS still demonstrated laterality (NAA/Cho and NAA/Cr ratios on the right side were lower than those on the left). Six months after embolization, these ratios on the right became closer to those on the left. Case 2: A 48 year-old female suffered from spontaneous CCF associated with cortical venous drainage. Before embolization of the CCF, ratios of NAA/Cho and NAA/Cr on the temporo-basal ganglia of the drainage side (left side) were lower than those on the contralateral side. After partial embolization of the CCF, which caused the angiographical disappearance of the cortical venous drainage, NAA/Cho and NAA/Cr ratios on the affected side increased to almost the same levels as those on the contralateral side. We consider that 1H-MRS is noninvasive and is a useful method to generate data evaluation of affected brain metabolism by venous reflux in cases of CCF associated with cortical venous drainage. PMID- 7477705 TI - [A case of gas-producing brain abscess with subdural empyema: timing of the operation for brain abscess]. AB - A case of gas-producing brain abscess with subdural abscess was reported. An 18 year-old boy was admitted with a five-day history of vomiting and high grade pyrexia. Plain skull roentgenograms demonstrated left frontal multiple gas bubbles. CT scan and MRI showed that both brain and subdural abscesses contained gas in the left frontal area. Antibiotics and glyceol were intravenously administrated. In serial CT scans, subdural abscess was not recognized, while brain abscess was enhanced in a ring. Seven days after admission, milky white pus with a fecal odor was aspirated using CT guided stereotactic apparatus, and the catheter was left in the abscess cavity. Culture of the pus grew peptostreptococcus. On the 22nd hospital day, repeated aspiration and drainage were performed for the residual brain abscess. The patient gradually improved after the aspiration with continuous administration of antibiotics, and he was discharged on the 59th hospital day with no neurological deficits. Based on our experience and a review of the literature, the treatment of choice is aspiration in the late cerebritis stage of brain abscess. PMID- 7477706 TI - [Recurrence of pleomorphic xanthoastrocytoma six years after total removal of mural nodule: a case report]. AB - A case of recurrent pleomorphic xanthoastrocytoma (PXA) was reported. A 46-year old male underwent total removal of mural nodule from cystic tumor in the left temporal lobe in 1986. Histological diagnosis was astrocytoma. There were no abnormal neurological signs or symptoms after the operation. However, 6 years after the operation he complained of strong unpleasant smell as of rotten onions and was admitted to our hospital. MRI revealed a cystic tumor with a mural nodule which was enhanced markedly by Gd-DTPA in the same location as previous tumor site. He was operated upon again and tumor was removed with the cyst wall. Histological diagnosis was PXA. Surgical procedures for removal of cystic PXA has been controversial; removal of tumor only, tumor removal with biopsy of cyst wall, or tumor removal with resection of cyst wall. Real reason for recurrence of this tumor in our case is unknown. However, since astrocytoma cells were found in the biopsy specimen of the cyst wall at time of the first operation, the tumor might have recurred from these cells. Therefore, this time we removed not only the mural nodule but also the cyst wall, because the cyst wall peeled off easily. We would like to propose that PXA cyst walls should be resected, whenever possible, in order to accomplish total removal of tumor. PMID- 7477708 TI - [Spinal multiple sclerosis mimicking a spinal cord tumor: a case report]. AB - Since the advent of magnetic resonance imaging (MRI), to visualize lesions of multiple sclerosis has become easy to do. However, in some cases with primary spinal cord multiple sclerosis, it is not always easy to obtain a diagnosis in the first instance. We reported a case of primary spinal multiple sclerosis diagnosed through histological examination of a surgical specimen taken by an open biopsy. A 35-year-old woman was admitted with complaints of two-months duration of progressive weakness and sensory disturbance in the legs and buttocks. On radiological examinations including metrizamide CT myelography and MRI, enlargement of the conus medullaris was the only positive finding. Respective to her clinical course, intramedullary spinal cord tumor could not be ruled out, so an open biopsy was performed. Histological examination revealed that the cord lesion was acute demyelination with perivascular inflammation. Her neurological signs were almost completely cured with administration of corticosteroid, though new brainstem signs took place two months later and then a concrete diagnosis of her having multiple sclerosis was finally achieved. Since preoperative examinations can not differentiate spinal cord tumor from any other intramedullary cord lesions such as demyelinating foci of multiple sclerosis, surgical intervention would be approved in such atypical primary spinal cord multiple sclerosis. PMID- 7477707 TI - [Primary malignant T-cell-rich B-cell lymphoma of the central nervous system: a case report]. AB - An autopsy case of primary intracranial T-cell-rich B-cell lymphoma in a 69-year old female is presented. The patient was admitted with a diagnosis of a brain tumor in July 1993 and a month long history of mental deterioration, motor weakness of the right arm and leg, and a tendency toward somnolence. Neurological examination revealed disturbance of consciousness, right hemiparesis, and papilloedema. However, her general physical examination was unremarkable. A CT scan and MR imaging revealed an irregular enhanced mass lesion at the paraventricular deep white matter in the bilateral parieto-occipital lobe. The patient was treated with surgical biopsy of the tumor followed by combined radiotherapy (a total of 50 Gy) and chemotherapy. Following repetitive episodes of remission and exacerbation, the patient expired about seven months after the onset of symptoms. Histopathological diagnosis of the tumor was malignant lymphoma (diffuse medium-sized cell type). In the immunohistochemical study, most of the lymphoma cells had T-cell markers, such as UCHL1. Some of the lymphoma cells were L26-positive. Neither glial fibrillary acidic protein nor neuron specific enolase were reactive with the lymphoma cells. At post-mortem examination, the specimens disclosed diffuse infiltration of medium-sized lymphoma cells. By contrast, most of the lymphoma cells were shown to be positive by the analysis of L26. None of the lymphoma cells exhibited the presence of UCHL1. These immunohistochemical evaluations conform to the criteria of T-cell rich B-cell lymphoma. PMID- 7477709 TI - [Spinal intradural arachnoid cyst associated with enlarged filum terminale]. AB - This is a report of a spinal intradural arachnoid cyst associated with an enlarged filum terminale. A 9-year-old female was admitted to our hospital complaining of lumbosacral pain. Neurological examination revealed no abnormal findings. CT scan showed a round shaped mass lesion in the right dorsolateral side of the spinal canal between the level of L1 to S1/2. The mass lesion disclosed isodensity and was not enhanced. A small low density mass lesion was visualized in the dural sac at the same level. MRI showed the mass lesion with low signal intensity on the T1-weighted image, and high signal intensity on the T2. The small mass lesion in the dural sac was demonstrated with low signal intensity on the T1 and T2-weighted image. In addition to this finding, Chiari malformation type I was disclosed. It was suspected that this mass lesion was extradural arachnoid cyst associated with an enlarged filum terminale. Osteoplastic laminotomy between L3 to S1/2 was performed, and an arachnoid cyst was found in the intradural space. It compressed the cauda equina and enlarged tough filum terminale. The cyst was removed subtotally, and the filum terminale was cut off. Postoperative course was uneventful and the lumbosacral pain disappeared. It was thought that this pain might be derived from the intradural arachnoid cyst, or the enlarged filum terminale.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7477710 TI - [De novo aneurysm associated with a persistent primitive trigeminal artery: case report]. AB - This paper presents a case of a de novo unruptured anterior communicating aneurysm associated with a persistent primitive trigeminal artery. The patient was a 65-year-old female with tinitis, for whom computed tomography with contrast medium indicated right dural enhancement. Dural arteriovernous malformation was suspected and angiography was thus conducted. A right persistent primitive trigeminal artery was demonstrated but no aneurysm could be found. Six years later, magnetic resonance angiography indicated an aneurysm on the anterior communicating artery. It was subsequently removed by surgery. A persistent primitive artery is frequently found to accompany aneurysm but de novo aneurysm associated with such an artery does not appear in the literature to date. The authors emphasize the importance of follow-up angiography in cases of persistent primitive artery not accompanied by an aneurysm. PMID- 7477711 TI - [Posttraumatic pseudoaneurysm of the middle meningeal artery: a case report]. AB - An epidural hematoma associated with posttraumatic pseudoaneurysm of the middle meningeal artery is a very rare clinical entity. Only 31 such cases have been reported in the literature. A 54-year-old man was admitted with lethargy and right hemiparesis. A skull X-ray film revealed a linear fracture in the left parietal and temporal bone, extending to the base of the skull. A brain CT scan disclosed an acute epidural hematoma over the left cerebral hemisphere, contusional hematoma in the right frontal lobe and thalamus, and subarachnoid hemorrhage in the left Sylvian fissure. During an emergency craniotomy a large epidural hematoma was evacuated and external decompression was performed. Angiography, which was performed about one month after the head injury, revealed an aneurysm of the middle meningeal artery. The aneurysm was removed to avoid delayed rupture at the time of cranioplasty. Histological diagnosis was pseudoaneurysm. We discussed the pathogenesis of a pseudoaneurysm of the meningeal artery and the necessity to pay attention to the presence of a pseudoaneurysm when an active bleeding point is not observed during surgery of the epidural hematoma. PMID- 7477712 TI - [A penetrating injury of the extracranial vertebral artery: a case report]. AB - Penetrating injury to the vertebral artery is a very rare accident and no case except ours has been reported to date in Japan. We reported a case of successful treatment of a penetrating injury by a nail to the left vertebral artery with the use of interventional radiologic procedure. A 27-year-old man was admitted to our hospital with a complaint of penetrating injury in his neck. Left vertebral angiography revealed that a nail had penetrated into the vertebral artery at the C6 level. Considering the possibility of thrombus formation around the metallic foreign body and the risk of uncontrollable bleeding, we performed embolization of the injured vertebral artery with Interlocking Detachable Coil. PMID- 7477713 TI - [A case of surgical retrieve of migrated microcatheter]. AB - This is a report of an accidental breakaway of a microcatheter during endovascular embolization for cerebral arteriovenous malformation occurring in a 48-year-old female. She had a sudden onset of cerebral herniated signs caused by a large hemorrhage in the region of the right parietal lobe. An angiogram showed a large arteriovenous malformation in the same region. 48 days after emergency evacuation of the hematoma and clipping of a feeding artery, we performed endovascular embolization using a microcatheter as assistant therapy for total removal of the nidus. However, accidentally, the microcatheter broke and migrated to the distal middle cerebral artery. Although we tried to retrieve the catheter using a loop snare, we had no success. We eventually retrieved it by open surgery next day. Fortunately the migrated catheter could be seen through a cortical artery, so after trapping it by using elastic thread and cutting off the artery we were able to extract the migrated catheter. Finally we were able to remove the nidus totally without other complications. Since breaking and migration of a microcatheter is a rare but possible accident, we should be prepared to cope with it appropriately. PMID- 7477715 TI - [Infantile brainstem infarction due to vertebral artery fenestration: a case report]. AB - The patient was a 10-year-old boy. He had a sudden onset of generalized tonic clonic convulsion. On admission, he presented with mild disturbance of consciousness and right hemiparesis. The electroencephalogram showed diffuse spike and slow wave complex. Three days later, magnetic resonance imaging (MRI) showed high signal intensity areas in the left cerebellar hemisphere and pons. On cerebral angiograms, coiling of the bilateral carotid arteries, occlusion of the basilar artery at the distal site, and fenestration of the left vertebral artery at the atlanto-axis level were shown. According to these findings, the patient was diagnosed with brainstem and cerebellar infarction. On the follow-up angiograms, recanalization of the basilar artery on the right vertebral angiogram, occlusion of the left vertebral artery at the distal site of the posterior inferior cerebellar artery origin, and occlusion of one of the duplicated vertebral arteries were recognized. Fenestration of the vertebral artery is presented in about 1-2% at angiography and autopsy and the clinical significance is controversial. In this case, the fenestration may have played a role as an embolic source because there was no probable cause of the cerebral infarction, and the vascular occlusion and recanalization occurred near the distal site of the fenestration. These findings suggest the clinical significance of vertebral artery fenestration as an embolic source. PMID- 7477714 TI - [A surgically treated case of ventrally exophytic pontine glioma]. AB - A surgically treated case of ventrally exophytic pontine glioma is reported. A 49 year-old woman, complaining of dysarthria, dysphagia and gait disturbance, was admitted to our department. Her past history included bronchial asthma. Plain skull x-p and tomography revealed destruction of the dorsum sellae and upper clivus. CT demonstrated an enhanced oval mass at the ventral side of the upper brainstem. The mass was severely compressing the brainstem dorsally. MRI revealed a low-intensity band between the tumor and the brainstem. Dynamic MRI demonstrated a pattern of rapid increasing and slow reduction. Cerebral angiogram demonstrated a paradoxical sign that pontine branches were located anterior to the basilar artery. The finding demonstrated that the tumor was an intraaxial mass. The first operation was performed by the orbitofrontomalar approach. On the trans-sylvian route, the tumor was removed partially with CUSA and neuronavigation system. Its histology was astrocytoma grade III. Radiation therapy was added. The patient's symptoms aggravated again. On the second operation, the transtemporal route with tentorial resection was chosen. Under a wide visual field, intracapsular subtotal resection of the tumor was performed. Interferon therapy was added. She was discharged to her home with no neurological deficits. Ventrally exophytic pontine glioma is very rare. Low-intensity band of MRI, a sign of extraaxial mass, was visualised in our case. On the other hand, cerebral angiogram demonstrated a paradoxical sign. This sign suggested that the tumor originated from the brainstem. With update skull base surgery and neuronavigation system, surgical therapy of ventrally exophytic pontine glioma is safe and effective. PMID- 7477716 TI - [Surgical techniques for aneurysms in and around the cavernous sinus]. PMID- 7477717 TI - [Intratumoral pharmacokinetics following intraarterial administration of MCNU in patients with malignant gliomas]. AB - This clinical study was undertaken to examine intratumoral (i.t.) pharmacokinetics after intraarterial (i.a.) administration of MCNU (80mg/m2) in 5 patients with glioblastoma (GB) and 2 with anaplastic astrocytoma (AA). After resection or stereotactic biopsy of the cystic lesion, an Ommaya reservoir was placed into the tumor cavity in all patients. The distribution of MCNU in blood was compatible with a two-compartment model, and the half life of the alpha-phase and beta-phase was 4.1 minutes and 160.4 minutes, respectively. MCNU was detected in the i.t. fluid in 5 cases, 4 of GB and 1 of AA. The concentration of i.t. MCNU gradually increased during the 5 to 30 minutes after i.a. injection to a level about 20.0% of its blood concentration. However, no MCNU was detected in patients showing partial response (3 of GB and 1 of AA) or no change (1 of GB) after the i.a. infusion of MCNU during maintenance chemotherapy. These results suggests that MCNU may transfer into the tumor tissues. Further investigation is warranted. PMID- 7477718 TI - [The effect on higher cortical dysfunction of percutaneous transluminal angioplasty for internal carotid artery stenosis]. AB - Seven patients with internal carotid artery (ICA) stenosis with higher cortical dysfunction due to hemodynamic ischemia were treated by percutaneous transluminal angioplasty (PTA). The patients ranged from 49 to 71 years of age, and included five males and two females. Neuropsychological tests were evaluated before and after PTA. Higher cortical dysfunction improved in all cases after PTA. It is concluded that PTA is effective to improve higher cortical dysfunction in patients who have ICA stenosis associated with hemodynamic compromise, if the ICA is satisfactorily dilated. PMID- 7477719 TI - [A case of multiple cerebral aneurysm which showed rapid growth caused by left atrial myxoma]. AB - A 24-year-old woman was admitted complaining of right hemiparesis and episodes of syncope. Computed tomography demonstrated a low density area in the left putaminal region. Intravenous digital subtraction angiography (IVDSA) showed two aneurysms in the distal segment of the right middle cerebral artery. Cerebral emboli from a cardiac source was suspected, and cardioechography was performed. Myxoma was located in the left atrium. The patient was transferred to a cardio surgical unit, and the myxoma was successfully removed. After removal by operation of the cardiac tumor, follow-up third IVDSA was performed. One aneurysm of the distal segment of the right middle cerebral artery had grown larger. On the other hand, the other aneurysm had disappeared. Clipping of the enlarged aneurysm was performed. After the clipping operation of the enlarged aneurysm, a follow-up 4th IVDSA was performed. A new aneurysm of the proximal segment of the left cerebral artery was observed. A follow-up 5th IVDSA was performed, revealing that the new aneurysm was enlarging. No operation was performed, because the aneurysm was the fusiform type. At present, the patient is complaining of slight right hemiparesis and has returned to her job. Here we reported a case of cerebral aneurysm caused by left atrial myxoma. PMID- 7477720 TI - [Ruptured mycotic aneurysm of the middle cerebral artery: a case report]. AB - A sixty-two-year-old woman was diagnosed as having the mitral valve insufficiency seven months prior to admission. The patient was admitted to the hospital with complaints of right hemiparesis and aphasia. CT scan revealed an intracerebral hematoma in the left front-parietal region. Cerebral angiography disclosed an aneurysm at the distal portion of left middle cerebral artery. An increase in the amount of C reactive protein and leukocytosis indicated the presence of inflammatory lesions. Antibiotics were administered because a mycotic aneurysm was suspected. White blood cell count and C reactive protein returned to normal after treatment with antibiotics for one months. The aneurysm had disappeared on the second angiography performed after treatment. Strategy for mycotic aneurysm is still controversial. 49 reported cases in the literature since 1975 were reviewed and the efficiency of antibiotic therapy was discussed. PMID- 7477721 TI - [Bacterial intracranial aneurysm associated with infective endocarditis: a case showing enlargement of aneurysm size]. AB - The authors report a case of bacterial intracranial aneurysm associated with infective endocarditis. A 48-year-old male was admitted on March 26, 1994, with complaints of difficulty in speaking and mild swelling of the right leg following mild fever. On examination he showed motor aphasia and mild weakness of the right upper and lower limbs. Cardiac auscultation revealed a grade 3/6 holosystolic murmur. Laboratory data revealed signs of infection through white blood cell count and CRP. Enterococcus faecalis was isolated from the blood culture at the time of admission. A computerized tomographic (CT) scan and magnetic resonance (MR) imaging showed a round mass with perifocal edema. Angiography revealed an aneurysm from the precentral artery of the left middle cerebral artery. A mycotic aneurysm due to bacterial endocarditis was diagnosed. The patient was treated with high doses of antibiotics. However, angiography 2 weeks after the initial study demonstrated the enlargement of the aneurysm and severe narrowing of the angular artery. On April 19, excision of the aneurysm was performed. Operative findings showed degeneration and thickening of the walls of the aneurysm. After the operation, antibiotic therapy was continued. The patient was asymptomatic upon discharge and has continued to do well. Repeated angiography on September 12 showed no further aneurysm. There is a danger of rupture in mycotic aneurysm due to bacterial endocarditis. It is important to repeat angiography and to manage the primary disease. If an aneurysm enlarges with serial angiography, it should be treated surgically without further delay. PMID- 7477722 TI - [A case of pituitary abscess caused by infection of Rathke's cleft cyst]. AB - Pituitary abscess is relatively rare. Only about 80 cases have been reported. Preexisting lesions in pituitary fossa, such as pituitary adenoma, craniopharyngioma and Rathke's cleft cyst, are inclined to be complicated by infection more than the normal pituitary glands are. We reported a case of pituitary abscess caused by infection of Rathke's cleft cyst. A 67-year-old male had general fatigue and loss of appetite 4 months before admission. On admission he was found to have diabetes mellitus, diabetes insipidus, and hypernatremia. These defects were controlled by medication but he gradually became comatose and febrile. CT and MRI revealed an intrasellar lesion with ring enhancement. Lumbar puncture demonstrated an increase of mononuclear cells and protein. Blood chemistry revealed a marked increase of CRP. He was operated on via the transsphenoidal approach, which revealed sphenoid sinusitis and abscess formation in the pituitary gland. Histological examination of the surgical specimen revealed infection of Rathke's cleft cyst but the fluid in the cyst was sterile. By the drainage of the cyst and the use of antibiotics the patient became alert and signs of infection disappeared. He was discharged with a slight hypopituitarism and returned to normal life. Mortality rate of pituitary abscess is decreasing but is still high because of hypopituitarism and severe infection. Accurate diagnosis and operation are necessary. Transsphenoidal surgery is preferable for postoperative drainage of the abscess. PMID- 7477723 TI - [A rare case of cranial metastasis from hepatocellular carcinoma]. AB - This is the first reported case of magnetic resonance imaging (MRI) findings of cranial metastasis from hepatocellular carcinoma. A 53-year-old male was admitted to our hospital on August 23, 1994, complaining of severe headache and a subcutaneous mass on the forehead. He was diagnosed as hepatocellular carcinoma in February, 1994, at another hospital. Because of multiple intrahepatic metastasis, he was inoperable and received transarterial embolization (TAE) on February 15, 1994. He had noticed the subcutaneous mass two months prior to admission, and its recent rapid growth, and morning headache. On admission, there was no abnormality observed by neurological and physical examination except the subcutaneous mass on his forehead, 5 x 7 cm in size. It was elastic soft and unmovable, and he felt tenderness. Laboratory examination showed only mild liver dysfunction. HBsAg was negative, and alpha-fetoprotein and PIVKA-II (Protein Induced by Vitamin K Antagonists) were within normal limit. Skull X-ray showed a round bone defect in the frontal bone. Computerized tomographic (CT) scan showed bone destruction and a well-circumscribed high density mass extending from the frontal subcutaneous region into the cranial cavity. MRI showed the tumor compressing the left frontal lobe on T1 weighted image as isointense and T2 weighted image showed a slight low intense mass. The tumor was clearly enhanced on both CT scan and MRI. Left external carotid angiogram demonstrated that the hypervascular tumor mainly fed by a frontal branch of the left superficial temporal artery in the frontal region. Tumor and bone scintigram revealed multiple bone metastasis. Lung CT scan showed no metastasis.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7477724 TI - Physiology and immunology of lymphatic drainage of interstitial and cerebrospinal fluid from the brain. PMID- 7477725 TI - Anatomical pathways for lymphatic drainage of the brain and their pathological significance. PMID- 7477726 TI - Imaging techniques for in vivo quantitation of extracranial lymphatic drainage of the brain. PMID- 7477727 TI - Focal brain damage enhances experimental allergic encephalomyelitis in brain and spinal cord. AB - The immunological basis of multiple sclerosis (MS) is well recognized but the factors inducing MS lesions are unclear. In this study, we test the hypothesis that focal brain injury, inflicted during the pre-clinical stages of experimental allergic encephalomyelitis (EAE), will enhance the severity of immunological damage in the cerebral hemispheres and spinal cord. Acute EAE was induced in 30 Lewis rats by the injection of guinea pig spinal cord homogenate in complete Freund's adjuvant. A cryolesion to the surface of the left cerebral hemisphere was induced at 3 days (n = 6) or 8 days (n = 10) postinoculation (p.i.) and animals were killed at 15 days p.i. Control animals were EAE only (n = 9), cryolesion only (n = 4), EAE and sham cryolesion (n = 5) and normal animals (n = 3). Brain and spinal cord were stained by immunocytochemistry using W3/13 (T lymphocytes) OX6 (MHC Class II) and GFAP (astrocytes) antibodies. The results showed a 2-fold increase in the number of EAE lesions in the brain with significant and widespread increase of MHC Class II antigen expression by microglia, in the cryolesion EAE 8 days p.i. when compared with EAE only animals. The pattern of enhancement suggests that it is due to (i) local spread of tissue or serum factors from the cryolesion; (ii) neural factors affecting remote regions of the CNS; (iii) stimulation of the immune system which may occur due to products of brain injury draining to regional cervical lymph nodes. Investigation of the mechanisms involved may prove fruitful in establishing factors which initiate, aggravate or ameliorate brain damage in multiple sclerosis. PMID- 7477728 TI - Single strand conformation analysis of two genes contained within the first intron of the neurofibromatosis type I gene in patients with multiple sclerosis. AB - Single strand conformation analysis has become the most widely used technique to screen large numbers of DNA samples for unknown mutations which may contribute to genetic susceptibility to disease. The method relies on the electrophoretic migration of a single strand of nucleic acid in a polyacrylamide gel being dependent on its conformation which is in turn dependent on its sequence. We have examined two closely related genes present in the first intron of the neurofibromatosis type I gene--the oligodendrocyte myelin glycoprotein (OMGP) gene and the ecotropic viral integration 2A (EVI2A) gene--in 36 patients with multiple sclerosis (MS) and 36 healthy controls. A single mutation was found in the OMGP gene which resulted in an amino-acid change of glycine to aspartic acid. This occurred in identical proportions (16.6%) in MS patients and controls. Two rare mutations were found in the EVI2A gene, one resulting in an arginine substituting for a glutamine (one control and one patient), the other in the replacement of a glycine with serine (one control only). A third common polymorphism was seen in 5'-untranslated region of the EVI2A gene, with 65% of patients and controls showing a T-->C transition in either a heterozygous or a homozygous form. This makes it unlikely that either of these genes is involved in genetic susceptibility to MS, but regions of these genes outside of the exonic sequences have not been examined. PMID- 7477729 TI - Apoptotic neurons in brains from paediatric patients with HIV-1 encephalitis and progressive encephalopathy. AB - The pathogenesis of human immunodeficiency virus type 1 (HIV-1) associated dementia in adults involves neuronal loss from discrete areas of the neocortex and subcortical regions, but the mechanism for neuronal death is poorly understood. Gene-directed cell death resulting in apoptosis is thought to be a normal feature of neuronal development, but little is known about neuronal apoptosis in disease states. We investigated whether HIV-1 infection of the central nervous system is spatially associated with apoptosis of neurons. Using an in situ technique to identify newly cleaved 3'-OH ends of DNA as a marker for apoptosis, we demonstrate the presence of apoptotic neurons in cerebral cortex and basal ganglia of children that had HIV-1 encephalitis with progressive encephalopathy. Furthermore, an association was observed between the localization of apoptotic neurons and perivascular inflammatory cell infiltrates containing HIV-1 infected macrophages and multinucleated giant cells. Apoptotic neurons and p24-positive macrophages were observed infrequently in cerebral cortex and basal ganglia in children with HIV-1 infection without encephalitis or clinical encephalopathy. In nine control (HIV-1 negative) brains, ranging from the first post-natal month of life to 16.5 years of age, infrequent neuronal apoptosis was observed in three cases. These findings suggest that neuronal apoptosis is unlikely to be associated with post-natal development except in early post-natal germinal matrix, and that it may instead represent the end result of specific pathological processes, such as HIV-1 encephalitis. PMID- 7477730 TI - Neuronal apoptosis in HIV infection in adults. AB - Productive infection of the central nervous system by HIV predominantly involves the white matter and basal ganglia. Involvement of the cerebral cortex with neuronal loss is also described in AIDS patients but not in asymptomatic HIV positive patients. The mechanism of neuronal damage is unknown. To enquire whether neuronal loss in AIDS may be due to an apoptotic process, we examined the cerebral cortex from 12 patients who died from AIDS using two different methods: in situ end labelling and gel electrophoresis of DNA to demonstrate DNA fragmentation. None of the patients had cerebral opportunistic infection or tumour. Four patients had no significant neuropathological changes, eight patients had variable cerebral atrophy and four of them also had productive HIV infection of the brain. These patients were compared with four HIV-positive asymptomatic patients, five seronegative asymptomatic controls, and two seronegative patients with Alzheimer's disease. We demonstrated neuronal apoptosis in the cortex in all AIDS patients, as well as in the Alzheimer's patients. Apoptosis was not observed in the asymptomatic cases whether seropositive or seronegative. Neuronal apoptosis was more severe in atrophic brains, and did not directly correlate with productive HIV infection, suggesting an indirect mechanism of neuronal damage is most likely. PMID- 7477731 TI - Experimental herpes simplex virus type 1 (HSV-1) infection of the spinal cord and dorsal root ganglia. AB - Host factors determining the outcome of herpes simplex virus type 1 (HSV-1) infection within neurons are poorly understood. This paper aims to identify regional differences in the behaviour of HSV-1 within the nervous system as an approach to investigating the role of the host environment in determining the outcome of infection. We describe a mouse model of HSV infection focused on motor neurons of the spinal cord, resulting from intramuscular injection (i.m.) and compare this with the behaviour of virus within sensory neurons following scarification of virus on to skin. Viral antigen was detectable immunohistochemically by 2 days in both models and disappeared by 9-11 days. The time course of acute infection was reflected in the i.m. group by quantitative plaque assay for virus. Inflammation and cell destruction occurred in both models, but clinical features and histological destruction were greater in the group infected via the intramuscular route. In the sensory ganglia, a latent state from which virus could be reactivated by explanation, was established with LATS expression detectable in many neurons at 35 days post-infection (p.i.), but not in non-neuronal cells. Expression of latency associated transcript (LATS) was detected in motor neutrons in spinal cords at 35 days p.i. providing evidence for establishment of a LATS-positive latent state at this site, and continued to be detectable up to 6 months post-infection. In addition, LATS was detected in white matter at late times, suggesting a non-neuronal site of latency. In contrast to the behaviour in sensory ganglia, induced reactivation from spinal cords, by explanation and nerve section, was a very rare event. We have shown that a LATS positive latent state can be established within motor neurons of the CNS, but that there are regional differences in the biology and outcome of infection between the CNS and peripheral nervous system. We propose that this may be a useful model to study reproducibly, the behaviour of HSV-1 in a CNS environment and, by comparison with sensory ganglion infection, to explore host factors which may underlie these regional differences. The relevance of this model for using HSV-1 as a therapeutic vector for motor neurons is also discussed. PMID- 7477732 TI - Grumose or foamy spheroid bodies involving astrocytes in the human brain. AB - Grumose or foamy spheroid bodies (GFSB), which correspond to the classical pathological description, 'grumose degeneration', are described. By light microscopy, GFSB are faintly eosinophilic and spheroidal structures with a foamy appearance in haematoxylin and eosin stains; they vary from 10 to 50 microns in diameter and contain amorphous debris-like material. Some GFSB, however, contain a varying amount of eosinophilic grumose aggregates, some of which are randomly stained with periodic acid Schiff (PAS), Schmorl, Berlin blue, Grimelius or silver methods. The Gallyas stain, on the other hand, usually stains the contents of GFSB black or brown. Immunohistochemically, most GFSB are ubiquitin-positive. Characteristically glial fibrillary acidic protein is associated with some GFSB giving a foamy appearance. Ultrastructurally, GFSB consist of dense bodies of various sizes and configuration. Glial fibrillary bundles and astrocytic punctate adhesions are occasionally observed associated with GFSB. Anatomically, GFSB are observed preferentially in the rostroventral parts of both the substantia nigra pars reticulata and the globus pallidus in a number of human neurodegenerative diseases and aged brains. GFSB, however, appear outside the above regions in various circumstances such as trauma, infarction and astrocytomas. In conclusion, GFSB are ubiquitinated structures closely related to astrocytes in their formation and with a preferred location in the deep regions of the basal ganglia. PMID- 7477733 TI - Neurodegeneration in the limbic and paralimbic system in progressive supranuclear palsy. AB - Progressive supranuclear palsy is neuropathologically characterized by neuronal degeneration of the basal ganglia, brain stem, and cerebellum. In addition, cortical neuronal degeneration associated with neurofibrillary tangles formation has been identified over wide areas of the brain in patients with progressive supranuclear palsy. We studied the distribution of alpha B-crystallin-positive degenerating neurons in cases with progressive supranuclear palsy, and compared them with those in Pick's disease. Alzheimer's disease, senile dementia of Alzheimer type, and normal aged individuals. A large number of alpha B-crystallin positive neurons was found in the cerebral cortices of four out of nine patients with progressive supranuclear palsy. In particular, alpha B-crystallin-positive ballooned neurons were frequently observed in deep cortical pyramidal cell layers of the limbic and paralimbic systems in these diseases. The involvement of the limbic and paralimbic systems may thus contribute to personality changes as well as to memory and cognitive impairment in some patients with progressive supranuclear palsy. PMID- 7477734 TI - Hereditary spastic paraparesis with dementia, amyotrophy and peripheral neuropathy. A neuropathological study. AB - Hereditary, probably autosomal recessive, spastic paraparesis in two siblings was associated with dementia of frontal lobe type, amyotrophy and peripheral sensory and motor polyneuropathy. Neuropathological findings correlate with neurological deficits, although neuron loss in the caudate and putamen, substantia nigra, and loss of Purkinje cells were clinically silent. Loss of neurons occurred in all cortical layers of the prefrontal lobe and superior temporal gyrus. Immunohistochemical studies showed reduced parvalbumin immunoreactivity in dendrites, and reduced numbers of calbindin D28k-immunoreactive cells, thus suggesting involvement of cortical local-circuit neurons. Myelin loss, ubiquitin immunoreactive granular deposits, and nerve fibre degeneration in the white matter of the frontal lobes and corpus callosum were also observed. Cerebral and subcortical white matter abnormalities, together with atrophy of the thalamic dorsomedial complex and anterior nucleus, may have accounted for the development of severe dementia in this patient. PMID- 7477735 TI - Brain stem nuclei in sudden infant death syndrome (SIDS): volumes, neuronal numbers and positions. AB - It has been suggested that the defect underlying the sudden infant death syndrome (SIDS) lies in brain stem nuclei involved in cardiac and respiratory function. However, most studies have not used rigorous quantitative techniques to assess brain stem nuclear volumes and neuronal numbers. We have measured the volume, neuronal numbers and position of brain stem nuclei in 11 SIDS and 11 aged-matched control infants. Using serial sagittal sections, nuclei involved in maintaining airway patency (hypoglossal, ambiguus and retroambiguus), heart rate (dorsal vagal) and generation of respiratory rhythm (ambiguus and dorsal vagal) were studied. No significant differences were found in nuclear volume increase with age, total neuronal number or nuclear position between SIDS and control cases. These findings support the hypothesis that the nervous system in SIDS may be normal until the final event that kills these infants. PMID- 7477736 TI - X-ray microprobe analysis of corpora amylacea. AB - Since corpora amylacea is concentrated in the high density fraction in the subcellular fractionation of autopsy brain. It is suspected that inorganic materials accumulate in corpora amylacea. Therefore, elemental analyses of partially purified corpora amylacea from autopsy brain from a patient with Alzheimer's disease and those from brain of a non-demented patient were performed by the X-ray microprobe method. Prominent peaks of sodium, phosphorus, sulphur and chloride were observed, and mapping analyses confirmed that these elements were actually contained within the corpora amylacea. A similar result was obtained using cryostat sections. Corpora amylacea are characteristically distributed along the margin of blood vessels, beneath the pial border of the hippocampus and in the subependymal zones of ventricles of aged brains, namely in the vicinity of blood and cerebrospinal fluid. From this distribution and from the results of the present paper, we suggest that corpora amylacea play a role in the absorption and accumulation of inorganic materials which have been extravasated from blood and cerebrospinal fluid (CSF) and taken up by astrocytes. This may reflect alteration of the blood-brain and blood-CSF barriers in the ageing brain. PMID- 7477737 TI - Microglia as a unique cellular target in the treatment of stroke: potential neurotoxic mediators produced by activated microglia. AB - The sequalae evoked by an initial ischemic event in the CNS are incredibly complex and involve a wide range of short-term and long-term metabolic adaptations. With regard to the 'penumbra' area, in which cell death occurs over a 1 to 3 day period, defining the roles of potential neurotoxic mediators is crucial. In this regard, upon cellular activation after an ischemic episode, microglia, the resident macrophages of the CNS, can produce large quantities of a number of neurotoxic mediators. These factors include excitatory amino acids, proteases, cytokinases and nitric oxides. In this manuscript, these mediators are reviewed with regard to the potential utility of suppression of microglial function, via immunosuppressive agents, in the treatment of stroke. PMID- 7477738 TI - Calpain as a novel target for treating acute neurodegenerative disorders. AB - Calpains are cytosolic, neutral proteases that normally exist in an inactive or quiescent state. They require higher than normal levels of calcium for activation which, once accomplished, lead to irreversible proteolysis of numerous cytoskeletal, membrane-associated and regulatory proteins. Because of these characteristics, calpain is gaining attention as a potentially important pathogenic variable in ischemic neuronal death. This manuscript explores this hypothesis by briefly reviewing current support for the role played by calpain in ischemic neurodegeneration, and then discussing a series of recently published studies which: 1. offer further evidence for the hypothesis, and 2. provide direct support for the idea that selective inhibition of calpain can greatly limit the neuronal damage that would normally occur following both global as well as focal brain ischemia. Thus, the data reviewed in this manuscript support the ideas that unregulated activation and proteolysis of intraneuronal calpain plays a significant role in the brain damage that occurs following an ischemic event and that delivering selective and membrane permeant calpain inhibitors to ischemic tissue may provide a powerfully effective therapeutic means of limiting neuronal damage. PMID- 7477739 TI - Therapeutic effects of endothelin receptor antagonists in stroke. AB - The presence of the endothelin isopeptides and endothelin receptors on neurons, glial cells and brain capillary endothelium suggests that endothelins may play a significant role in neuromodulation, astrocytic function and in regulation of cerebral blood flow. Furthermore, endothelins may play a significant role in the central regulation of neuroendocrine and autonomic nervous system functions (i.e., plasma volume, cardiovascular and respiratory control). Endothelin has potent cerebrovascular and proliferative effects suggesting a pathogenic role in cerebrovascular diseases. Endothelin receptors may represent important therapeutic targets for the treatment of both hemorrhagic and ischemic stroke. A review of the available data on endothelin levels and the effects of endothelin antagonists in cerebrovascular diseases is provided in the present report. Most notably is evidence in support of increased brain endothelin levels in hemorrhagic and ischemic stroke both in animal models and in humans. Also, endothelin receptor antagonists exert significant efficacy in animal models of cerebrovascular disease. For example, SB 209670, a rationally designed, potent, nonpeptide endothelin receptor antagonist, exerts therapeutic efficacy in reducing vasospasm following subarachnoid hemorrhage and neuroprotection following ischemic stroke. Certainly the available data warrants further evaluation of novel, selective endothelin receptor antagonists or endothelin converting enzyme inhibitors in cerebrovascular diseases. PMID- 7477740 TI - Neuroprotective actions of 1-aminocyclopropanecarboxylic acid (ACPC): a partial agonist at strychnine-insensitive glycine sites. AB - 1-Aminocyclopropanecarboxylic acid is a high affinity ligand with partial agonist properties at strychnine-insensitive glycine sites associated with the N-methyl-D aspartate subtype of glutamate receptors. Since occupation of these sites appears required for operation of N-methyl-D-aspartate, receptor coupled cation channels, it was hypothesized that a glycine partial agonist could function as an N-methyl D-aspartate antagonist. This hypothesis was examined by evaluating the in vivo and in vitro neuroprotective actions of 1-aminocyclopropanecarboxylic acid. 1 Aminocyclopropanecarboxlic acid (150-600 mg kg-1) administered to gerbils five minutes following twenty minutes of forebrain ischemia significantly improved seven day survival; the optimal dose (300 mg kg-1) increased 7 days survival > 4 fold, from 20% to 92%. Survival of hippocampal CA1 neurons (quantitated 7 days post-ischemia) was significantly (approximately 3-fold) increased by the 600 mg kg-1 dose. Seven day survival was not significantly increased when the interval between reperfusion and drug administration (300 mg kg-1) was increased from 5 to 30 min. In cerebellar granule cell cultures, NMDA combined with a saturating concentration of glycine (10 microM) resulted in a 500% increase in cGMP levels. cGMP levels were increased by 100% over basal when NMDA was combined with a saturating (10 microM) concentration of ACPC, indicating that in this measure, the efficacy of ACPC relative to glycine was approximately 0.2. Consistent with previous findings, 1-aminocyclopropanecarboxylic acid significantly reduced glutamate-induced neurotoxicity in cerebellar granule cell cultures. ACPC was most effective in blocking neurotoxicity at glutamate concentrations producing low to moderate levels of cell death.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7477741 TI - Diffusion weighted imaging for acute cerebral infarction. AB - The diagnosis of acute cerebral ischemia remains difficult. Computed tomography and magnetic resonance tomography are not specific at early time points. Diffusion imaging a new technique for magnetic resonance imaging was introduced in experimental studies and shown to have much earlier sensitivity for acute ischemic infarction. Diffusion imaging demonstrated early hyperintensities within minutes after ischemic onset and correlated well with post-mortem studies of the final infarct size. On the basis of molecular motion of water, hyperintensities during ischemia on diffusion imaging are most likely related to the development of ischemia induced cytotoxic edema and to slower diffusional motion. Experimental treatment and reperfusion studies with diffusion imaging demonstrated lesion reversal in vivo. Continuous monitoring after induction of ischemia enables an in vivo demonstration of the evolution of the ischemic lesion. Quantitative analysis of the apparent diffusion coefficient may enable the differentiation of infarcted tissue in the core of the lesion from reversibly damaged tissue. Preliminary clinical studies demonstrated a superior sensitivity of diffusion imaging compared with T2-weighted imaging. Clinicians may be able in the near future to image early stroke and to assess therapy when the stroke is still evolving. PMID- 7477742 TI - Development of HU-211 as a neuroprotectant for ischemic brain damage. AB - HU-211 (Dexanabinol), a putative neuroprotective agent, was evaluated in a number of animal models including closed head injury, optic nerve crush, global ischemia and focal ischemia. In these models, a single injection of HU-211 given after the insult confers long term functional improvement and significant increase in neuronal survival. Neuroprotective doses of HU-211 were found to be safe in a 14 day toxicological study in two species. In terms of mechanism, the compound behaves as a noncompetitive NMDA antagonist in vitro and in vivo. It blocks NMDA stimulated calcium influx in primary neuronal culture, and head injury related calcium influx in rats. In addition, HU-211 is a potent scavenger of peroxy and hydroxy radicals in vitro and it protects cultured neurons from toxicity of radical generators. Thus, Dexanabinol holds a unique position among putative neuroprotective agents since it combines NMDA blocking activity and free radical scavenging properties in one molecule. These observations support the development of HU-211 as a novel, multiple-action treatment approach for brain damage associated with stroke, cardiac arrest and trauma. PMID- 7477743 TI - Citicoline (CDP-choline): mechanisms of action and effects in ischemic brain injury. AB - Citicoline is approved in Europe and Japan for use in stroke, head trauma and other neurological disorders. It is presently being evaluated in phase II/III stroke trials in the United States. Exogenous administration of CDP-choline provides both choline and cytidine which access the brain and serve as substrates for the synthesis of phosphatidylcholine, a primary neuronal membrane component; the choline also enhances brain acetylcholine synthesis. Membrane repair and regeneration is necessary for recovery from stroke. Furthermore, citicoline may alleviate free fatty acid-induced toxicity which accompanies ischemic insult. Data from many pre-clinical and clinical trials support the hypothesis that citicoline may be a safe and effective treatment for stroke. PMID- 7477744 TI - Nitric oxide synthase, immunophilins and poly(ADP-ribose) synthetase: novel targets for the development of neuroprotective drugs. AB - During ischemic stroke, massive neural damage occurs due to excess release of glutamate which acts mainly through N-methyl-D-aspartate (NMDA) receptors. Activation of the NMDA receptor stimulates nitric oxide (NO) production by NO synthase (NOS). NO mediates glutamate neurotoxicity as inhibitors of NOS prevent neuronal death. FK506, an immunosuppressant drug, binds to FK506 binding protein (FKBP). One target of the FK506/FKBP complex is the calcium/calmodulin-dependent protein phosphatase calcineurin, whose activity is inhibited upon interaction with FK506/FKBP. FK506 treatment increases phosphorylation level of calcinurin substrates including NOS. As a potent neuroprotective agent in vitro and in vivo, FK506 increases NOS phosphorylation and decreases NO production. NO activates poly(ADP-ribose) synthetase (PARS), a nuclear enzyme that synthesizes poly(ADP ribose) from NAD. Prolonged activation of PARS depletes NAD and lowers cellular energy levels. Inhibition of PARS also prevents NO toxicity. NOS inhibitors, immunosuppressants and PARS inhibitors may be useful agents to prevent neuronal damage during stroke. PMID- 7477745 TI - Intraoperative radiation therapy in malignant glioma: early clinical results. AB - Intraoperative radiation therapy (IORT) with high energy electron beams is a treatment modality that has been included in multimodal programs in oncology to improve local tumor control. From August 1991 to December 1993, 17 patients with primary (8) or recurrent (9) high grade malignant gliomas, anaplastic astrocytoma (4), anaplastic oligodendroglioma (6) and glioblastoma multiforme (7), underwent surgical resection and a single dose of 10-20 Gy intraoperative radiation therapy was delivered in tumor bed. Fourteen patients received either pre-operative (8) or post-operative (6) external beam radiation therapy. Primary gliomas: 18-months actuarial survival rate has been 56% (range: 1-21+ months) and the median survival time has not yet been achieved. Four patients developed tumor progression (median time to tumor progression: 9 months). Recurrent gliomas: 18 months actuarial survival rate and median survival time has been 47% and 13 months (range: 6-32+ months) respectively. The median time to tumor progression was 11 months. No IORT related mortality has been observed. IORT is an attractive, tolerable and feasible treatment modality as antitumoral intensification procedure in high grade malignant gliomas. PMID- 7477746 TI - Modified solution for filling MRI coordinate indicators for the ZD stereotactic frame. AB - Increased use of magnetic resonance imaging and magnetic resonance angiography in stereotactic planning highlight some technical problems in imaging. Visualization of markers during imaging are vital in stereotactic planning. In magnetic resonance imaging-guided stereotaxy, specially designed plates are used that contain water-based copper sulfate solutions as a marker. Replacement of copper sulfate solution is usually required after some time due to precipitation and leaking. The refilling procedure is quite cumbersome and can cause technical problems in imaging. In this study, we describe copper sulfate solution mixed with a water-soluble gel as a simple but useful alternative to water-based copper sulfate resolution to ease the refilling procedure and to overcome the air bubbles. PMID- 7477747 TI - The effects of a carotid-jugular fistula on cerebral blood flow in the cat: an experimental study in the chronic period. AB - The purpose of the present study was to investigate the effects of feline carotid jugular fistula (Spetzler's model) on cerebral blood flow at 8 weeks after creation of the fistula. Using laser-Doppler flowmetry, cortical cerebral blood flow (CBF) on the fistula side was measured in 10 cats, Occlusion of the fistula showed no cortical CBF changes in 6 animals, but transient increases in 4. CO2 reactivity in the closed fistula was preserved. No neurological or histological abnormalities were recognized. These findings suggested that cerebral hemodynamic changes due to Spetzler's CJ fistula model are minimal even in the chronic period after fistulization. Therefore, creation of another experimental arterio-venous fistula model is mandatory for the investigation of hemodynamic changes following resection of a cerebral arteriovenous malformation. PMID- 7477748 TI - Protection by Iloprost (stable analogue of prostacyclin) of endothelial damage due to chronic vasospasm in dogs: an electron microscope study. AB - The resolution of cerebral vasospasm and protection of endothelial damage by Iloprost was evaluated with multicisternal injections. Sixteen adult mongrel dogs (18-20 kg) were assigned to one of three experimental groups. All animals received a total amount of 15 ml fresh unheparinized arterial blood via three injections into the cisterna magna. Selective vertebral angiography was performed on day 0 and subsequently blood injections were performed on the 2nd and 3rd days after the first injection. On the 7th day angiography was reperformed to determine the chronic vasospasm. The first group (5 dogs) was the control group and received intrathecal saline which was equal to the amount of saline in which Iloprost was diluted. The second group (5 dogs) did not receive any treatment until the 7th day. The third group (6 dogs) received Iloprost intrathecally (total 10 micrograms kg-1). In the first two groups angiographic vasospasm was prominent. For the second group intraarterial Iloprost was given on the 7th day in order to evaluate its acute effect. However there was no evidence of resolution of vasospasm. In the third group, resolution of vasospasm was verified on angiograms. Electron microscope studies of basilar arteries of the first two groups revealed degenerative changes of the endothelial cells which were separated from each other and the elastic lamina was irregularly arranged. In the intrathecal Iloprost-treated group there was little thickening in the elastic lamina and the endothelial cells were almost normal in structure. These results can be considered as the evidence of the prophylactic effect of Iloprost given by the intrathecal route in the prevention of chronic cerebral vasospasm. PMID- 7477749 TI - Mechanism research of cryoanalgesia. AB - The sciatic nerves of rabbits were frozen at different temperatures (-20 degrees C, -60 degrees C, -100 degrees C, -140 degrees C, and -180 degrees C). The morphology and function of the frozen nerves were examined with light microscopy (hematoxylin and eosinophilin stain and a histochemical thiocholine method) and electron microscopy. The function of the nerve after freezing was assessed using short latency somatosensory evoked potentials, sensory conduction velocity, and electromyogram at various intervals after freezing. There were no changes in morphology or function of nerves cryolesioned at -20 degrees C. The nerve fibers cryolesioned at -60 degrees C showed signs of freezing degeneration and lost their conductive function although, these nerves all recovered. Approximately half of nerve fibers cryolesioned at -100 degrees C showed Wallerian degeneration, and although the time to remyelination was delayed, nerve regeneration was still complete. At -140 degrees C and -180 degrees C the nerve fibers showed immediate necrosis, with destruction of basal membranes and proliferation of collagen fibers. The results explained the mechanism of cryoanalgesia. Our study demonstrates that cryo-temperatures lower than -140 degrees C will cause permanent alterations in nerve morphology and function, whereas warmer temperatures do not result in permanent nerve damage and are therefore not likely to provide long-term analgesia to patients. PMID- 7477750 TI - Fetal striatal homotransplantation for Huntington's disease: first two case reports. AB - Based on the successful use of fetal striatal brain grafting in the restoration of striatal function in rat and nonhuman primate models of Huntington's disease, as well as on the evidence for the clinical potential of fetal brain grafting in the treatment of Parkinson's disease, homotopic fetal striatal homotransplantations were performed in two huntingtonians. Case 1 was a 37 year old female with moderate to severe Huntington's disease of 9 years evolution; case 2 was a 29 year-old male with mild Huntington's disease of 5 years evolution. Using open microsurgery, each patient was implanted to the ventricular wall of the right caudate nucleus with both striata from a 13 week-old and a 12 week-old human fetus, respectively. Since surgery both patients were kept on cyclosporine A. Surgery produced no damaging effect to either patient. The time course of the neurological progression of their disease, spanning 33 months for case 1, and 16 months for case 2, reveal that the disease in both patients has progressed more slowly in relation to their preoperative state. Although presently it is not possible to determine to what extent, surgery has modified the course of their disease, or if it will continue to have an effect on it, these surgeries represent the first step towards the development of brain grafting for Huntington's disease. PMID- 7477751 TI - Plexus neuropathy following vaccination against tick-borne encephalitis and tetanus due to a sports related altered immune state. AB - A 45-year-old female patient who was practising sports on a competition level, developed a plexus neuropathy subsequent to a vaccination against tick-borne encephalitis (TBE) followed by an inoculation of tetanus toxoid six days later. After the TBE vaccination she continued to exercise intensely until symptoms of a neuritis were noticed. Pronounced endurance exercise has been described to alter the composition and function of the immune system. In our patient a decrease of T helper cells and a significant lowered CD4/CD8 ratio could be detected. The possible link between an altered immune state and post-vaccinal neuropathy is discussed. PMID- 7477752 TI - Lissencephaly and other malformations of cortical development: 1995 update. AB - Neuronal migration disorders are a group of malformations of the brain which primarily affect development of the cerebral cortex. The best known of these is lissencephaly (smooth brain). Most types result from incomplete neuronal migration to the cortex during the third and fourth months of gestation. In this review, we describe and illustrate the different types of neuronal migration disorders. We also review the many different genetic syndromes associated with neuronal migration disorders. Over 25 syndromes with lissencephaly or other neuronal migration disorders have been described. Among them are syndromes with several different patterns of inheritance including chromosomal or new mutation autosomal dominant, autosomal recessive, X-linked and unknown. Genetic counseling thus differs greatly between syndromes. The genes responsible for several of the lissencephaly syndromes have been mapped. X-linked lissencephaly has tentatively been mapped to chromosome Xq22 based on observation of a single X-autosomal translocation in a girl. Both Miller-Dieker syndrome and isolated lissencephaly sequence (in many patients) were mapped to chromosome 17p13.3 by detection of deletions and other structural chromosome rearrangements. Fukuyama congenital muscular dystrophy was mapped to chromosome 9q31-33 by homozygosity mapping. PMID- 7477753 TI - Preserved merosin M-chain (or laminin-alpha 2) expression in skeletal muscle distinguishes Walker-Warburg syndrome from Fukuyama muscular dystrophy and merosin-deficient congenital muscular dystrophy. AB - The merosin M-chain (or laminin-alpha 2) is one of three subunits of laminin-2 which is highly expressed in striated muscle and peripheral nerve. Complete lack of laminin-alpha 2 expression in skeletal muscle is the hallmark of one form of congenital muscular dystrophy which is characterized by dysmyelination of the central nervous system (CNS), links to chromosome 6q2 and is common among Caucasians. Laminin-alpha 2 expression was also found to be significantly reduced in Fukuyama congenital muscular dystrophy which links to chromosome 9q3. We report consistently preserved laminin-2 expression, including laminin-alpha 2, as detected by immunofluorescence in skeletal muscle from five patients with Walker Warburg syndrome which is characterized by congenital muscular dystrophy and, in addition, type II lissencephaly or pachygyria, defective CNS myelination, and ocular dysgenesis. These findings show that in spite of partial phenotypic overlap between Fukuyama CMD and Walker-Warburg syndrome the two disorders are nosologically separate disease entities. They also exclude that Walker-Warburg syndrome is allelic to the common form of congenital muscular dystrophy with laminin-alpha 2 deficiency. PMID- 7477754 TI - Minor neurological and perceptuo-motor deficits in children with congenital muscular dystrophy: correlation with brain MRI changes. AB - Diffuse white matter changes on magnetic resonance imaging (MRI) have been a consistent feature in some children with the "pure" form of congenital muscular dystrophy (CMD) in which there are no structural changes in the brain or severe mental retardation. The aim of this study was to assess fine motor and perceptuo motor abilities in children with CMD with and without MRI changes. Twenty-two children with "pure" CMD were investigated with a standard neurological examination and a battery of tests (Manual dexterity from the Movement ABC, test of visual-motor integration, Zurich Neuromotor test) which have already been used to detect minor neurological signs related to white matter changes. The cohort was then divided in two groups for analysis depending on the presence or the absence of diffuse white matter changes. A significant difference was found for all the tests between the group of the CMD children with normal MRI and the group with diffuse white matter changes. The manual dexterity and the Zurich Neuromotor tests showed a greater sensitivity than the test of visual-motor integration, which had some false negatives. It is of interest that in the group with diffuse white matter changes the presence of contractures or weakness did not seem to affect the quality of the performance; all these children scored abnormally on the test, irrespective of the severity or the extent of contractures and weakness. In contrast, in children with normal MRI severe contractures and weakness did affect the performances. Our results demonstrate that perceptuo motor difficulties and minor neurological soft signs are a consistent feature in CMD children with diffuse MRI changes but not with normal MRI.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7477756 TI - Estimation of critical closing pressure in the cerebral circulation of newborns. AB - We have developed a method to estimate the critical closing pressure (CrCP) of the cerebral circulation based on the intrinsic variability of arterial blood pressure (BP) around stable values of mean arterial pressure (MAP). A consecutive cohort of 33 premature newborns was studied at 6, 12, 24, 48 and 72 hours of age. Cerebral blood flow velocity (CBFV) was measured with Doppler ultrasound in the middle cerebral artery and BP was recorded in the abdominal aorta or in a peripheral artery. Continuous measurements lasting five minutes were recorded on digital magnetic tape and signals were digitized at a rate of 200 samples/seconds for processing on a digital computer. Mean values of BP (mBP) and CBFV (mBV) were computed for each cardiac cycle and CrCP was determined as the pressure axis intercept of the regression line of mBV as a function of mBP using 100 sequential heart beats. The resistance-area product (RAP) was obtained from the slope of the regression line. For 57 records (30 patients) the mean +/- SD values of CrCP and RAP were 23.9 +/- 11.6 mmHg and 4.07 +/- 1.83 x 10(4) kg.m-2.s-1, respectively. CrCP has a highly significant correlation with mean arterial pressure (p < 0.001) but RAP has not. Neither CrCP nor RAP are significantly correlated (p > 0.05) with PO2, PCO2, pH, haematocrit, gestational age, birthweight, postnatal age, heart rate on Pourcelot's resistance index. Our results suggest that cerebral perfusion pressure should be defined as MAP-CrCP for normal values of intracranial pressure. PMID- 7477755 TI - An X:autosome translocation stabilizes truncated dystrophin: implications for lack of truncated dystrophins in Duchenne muscular dystrophy. AB - We report a 5-year-old girl with clinical symptoms of typical Duchenne muscular dystrophy in males. The girl showed dramatic elevations of serum creatine kinase, and muscle biopsy histopathology consistent with a severe dystrophic myopathy. Cytogenetic analysis revealed an X:22 translocation (46,X,t [X;22] [p21.2;11.2]). Dystrophin immunofluoresence studies showed strong membrane immunostaining of dystrophin with antibodies directed against the amino terminus of the protein, but vastly reduced immunostaining with carboxyl-terminal antibodies. Immunoblot studies showed a major immunoreactive protein of approximately 350 kDa at approximately 20% levels. Nested RT-PCR analysis of the dystrophin mRNA in the patient's muscle showed the RNA to be positive for primers covering the first 85% of the dystrophin coding sequence, and negative for the carboxyl-terminal 15%. Taken together, our data suggests that the translocation breakpoint occurs towards the 3' end of the gene. The translocated dystrophin gene is still expressed into a truncated dystrophin protein associated with the plasma membrane. Our results are consistent with the translocation resulting in a more stable abnormal dystrophin mRNA. PMID- 7477757 TI - Retinal vasculitis caused by rubella. AB - A patient with vasculitis apparently caused by rubella and diagnosed during her life-time is here presented. The girl showed optic neuritis and large vascular changes in the eye fundi (retinal vasculitis). High intrathecal antibody production against rubella but little or none against other viruses was found in the cerebrospinal fluid. This study suggests that immunoactivation after rubella vaccination may remain and lead to vascular changes. PMID- 7477758 TI - Fatal tumor hemorrhage after ventriculoperitoneal shunting. Case report. AB - We report a fatal tumor hemorrhage in a 1-and-a-half-year old girl after ventriculoperitoneal shunting for obstructive hydrocephalus caused by a thalamic tumor. The possible pathophysiological mechanism is briefly discussed and the literature is reviewed. PMID- 7477759 TI - Current bibliographies of neuropeptides prepared by the University of Sheffield Biomedical Information Service. PMID- 7477760 TI - Comparison of soluble and membrane-bound pyroglutamyl peptidase I activities in rat brain tissues in the presence of detergents. AB - Pyroglutamyl peptidase I activity from soluble and membrane-bound fractions of rat brain homogenates is inhibited by the presence of sodium deoxycholate but not by triton X-100. Biobeads SM2, a polystyrene adsorbent reported to be useful in removing detergents from aqueous solutions, inhibits enzymatic activity in both fractions regardless of the presence of these detergents, probably because of partial adsorption of the enzyme by the polymeric microspheres. These effects seem to be enzyme-specific since other aminopeptidase activities are not affected by detergents or biobeads. The results suggest that soluble and membrane-bound forms of the enzyme represent the same protein in two different cell compartments. PMID- 7477761 TI - Acute and subchronic effects of dopamine agonists on neuropeptide gene expression in the rat striatum. AB - Acute and subchronic (7 days) effects of subcutaneous administration of the D-1 agonist (SKF 38393; 5.0 mg/kg), the D-2 agonist (quinpirole; 5.0 mg/kg), the mixed D-1/D-2 agonist (apomorphine; 5.0 mg/kg) or pergolide (0.5 mg/kg) on enkephalin and substance P gene expression in the striatum were studied in rats using in situ hybridization. Striatal enkephalin mRNA levels were unaffected by acute dopamine agonist treatment despite animals exhibiting altered motor behaviour. Only acute apomorphine treatment increased substance P mRNA (25%, p < 0.01). Following subchronic administration of pergolide, enkephalin mRNA expression was decreased by 23% (p < 0.05) while other drugs were without effect. The abundance of striatal substance P mRNA was increased only following subchronic pergolide treatment (35%, p < 0.05). These data suggest that peptide gene expression in the intact striatum is relatively resistent to intermittent dopamine agonist stimulation in contrast to the pronounced alterations previously observed in 6-hydroxydopamine lesioned rats or following blockade of dopamine receptors. PMID- 7477762 TI - Characterization of the opioid receptor subtypes mediating the negative inotropic effects of DAMGO, DPDPE and U-50, 488H in isolated human right atria strips. AB - The present investigation was aimed at elucidating if mu-, delta- and kappa opioid receptors are involved in the effects of DAMGO (selective mu-agonist), DPDPE (selective delta-agonist) and U-50,488H (selective kappa-agonist) in isolated electrically driven human right atria strips. The negative inotropic effects induced by the opioid agonists used in this study were not antagonized in presence of naloxone (preferentially mu-antagonist), naltrindole (selective delta antagonist) and norbinaltorphimine (selective kappa-antagonist). These data suggest that the opioid receptors are not involved in the cardiac depressant effects induced by mu-, delta- and kappa-opioid agonists. PMID- 7477763 TI - Vasodilation in porcine ophthalmic artery: peptide interaction with acetylcholine and endothelial dependence. AB - Co-activation of cranial perivascular sensory and parasympathetic fibres in vivo induces simultaneous release of several vasodilatory substances with neurotransmitter or neuromodulatory roles. The role of the endothelium and possible interactions between such substances are poorly understood. The objective of this study was therefore to investigate these aspects with the sensory dilator calcitonin gene-related peptide (CGRP) and the parasympathetic dilators acetylcholine (ACh) and vasoactive intestinal peptide (VIP) in isolated porcine ophthalmic artery. Whilst ACh induced relatively rapid, endothelium dependent dilation, CGRP and VIP induced slower dilations. Both CGRP and VIP were found to have partial endothelium-dependence in this artery. The simultaneous addition of ACh with CGRP potentiated the relaxation induced by CGRP, as has already been shown for substance P. ACh did not potentiate VIP relaxation, but the results generally indicate a potential role for ACh in initiating rapid dilation prior to strong, sustained relaxation by CGRP or VIP. The potential role of the endothelium and of substances like ACh or substance P in enhancing the rate of dilation of neuropeptides inducing strong and sustained relaxation is discussed. PMID- 7477764 TI - Somatostatin (SRIF) stimulates the release of interleukin-6 (IL-6) from human peripheral blood monocytes (PBM) in vitro. AB - Recent evidence has revealed that various neuropeptides appear to have distinct roles as immunomodulators. The aim of this study was to evaluate the role of hypothalamic neuropeptides (thyreoliberin [TRH], somatostatin [SRIF], and gonadoliberin [LH-RH]) on the secretion of interleukin-1 beta (IL-1 beta) and interleukin-6 (IL-6) from lipopolysaccharide (LPS) activated human peripheral blood monocytes (PM) cultured in vitro. LPS in concentration 1.5 micrograms/ml stimulated PBM to release IL-1 beta or IL-6 into the supernatants. SRIF in concentrations from 10(-8)M to 10(-10)M (but neither RH nor LH-RH in the same concentrations) potentiated the release of IL-6 from PBM. None of the tested neuropeptides stimulated the release of IL-1 beta from LPS activated human monocytes. These data indicate that SRIF in physiological or pharmacological concentrations which activate the release of IL-6 from PBM may be one of the regulators of immune response in humans. PMID- 7477765 TI - Effects of two newly synthetized cholecystokinin tetrapeptides on the activity of single hippocampal and thalamic neurons. AB - The influence of two iontophoretically administered newly developed cholecystokinin (CCK) tetrapeptides with high selectivity and affinity to CCK-B receptors on the impulse activity of single hippocampal and thalamic neurons were tested in in-vivo experiments, in comparison to the effect of the sulfated octapeptide (CCK8S). A very similar responsiveness to the compared drugs was found. Most neurons responded with an increase of their discharge frequency. Only a few suppressive effects were elicited by each drug and in each of the structures. There was a good correspondence between the compared drugs concerning the direction and relative response amplitude, resulting in a highly significant correlation of the effects of both CCK4s with the CCK8S effects. On a subsample of neurons, the blocking effect of the selective CCK-B receptor blocker PD135 was tested and found to be effective in 16 out of 20 CCK4 responses, including also one inhibition. The results show that the new compounds act as effective CCK agonist binding to the B-type CCK receptor. The few inhibitory effects obtained could be explained by possible indirect effects mediated via inhibitory interneurons which are known to exist in both investigated structures. PMID- 7477766 TI - Rat meningeal fibroblasts in primary culture express the proenkephalin gene. AB - When brought into primary culture, rat meningeal fibroblasts contained proenkephalin-mRNA detected with Northern blot hybridization. In contrast, splenic fibroblasts did not express the gene under the same culture conditions. In situ hybridization showed that the meningeal fibroblasts did not uniformly express the gene: groups of positive cells were surrounded by cells with low or no proenkephalin-mRNA. Some fibroblasts which contained the mRNA species took up bromo-deoxyuridine indicating that the expression of the gene also occurred in proliferating cells, but was not restricted to this group. In chromaffin and astroglial cells, activation of protein kinase A or C with 8 Br.cAMP or O tetradecanoyl 13-phorbolacetate, respectively, increases the expression of the gene. In meningeal fibroblasts, however, both agents reduced the levels of proenkephalin-mRNA. In the case of 8Br.cAMP, this effect was blocked by the protein synthesis inhibitor cycloheximide indicating that a newly-synthesized protein was involved. Cultured meningeal fibroblasts appear to be useful for studies on the cell specificity of the expression of this peptide gene as well on its regulation. PMID- 7477767 TI - Plasma pancreastatin-like immunoreactivity correlates with plasma norepinephrine levels in essential hypertension. AB - Pancreastatin (PST), a 49 amino acid peptide originally isolated from porcine pancreas, is derived from chromogranin A (Cg A), an acidic protein co-released with catecholamines from sympathetic nerve terminals and chromaffin cells. Extracellular processing of Cg A yields PST as well as other biological active peptides. Measurement of Cg A and PST-like immunoreactivity (PST-LI) has been used to investigate patients with pheochromocytoma and other neuroendocrine neoplasia. Some studies have found increased plasma norepinephrine (NE) levels in essential hypertension. We therefore measured venous plasma PST-LI and catecholamines in patients with essential hypertension. We employed a radioimmunoassay developed with commercially available reagents for measuring plasma PST-like immunoreactivity, and HPLC with electrochemical detection for measurement of plasma catecholamines. The correlation of PST-LI with epinephrine (E) was very weak. However, its correlation with NE was highly significant. Thus, venous plasma PST-LI immunoreactivity may reflect sympathetic nerve activity in essential hypertension. PMID- 7477768 TI - The role of growth factor receptors in central nervous system development and neoplasia. AB - Future advances in neuro-oncology will increasingly rely on an understanding of the molecular biology of brain tumors. Recent laboratory work, including the identification of oncogenes and tumor suppressor genes, has elucidated many of the molecular events contributing to oncogenesis. In particular, the signaling pathways for the growth factors have been implicated in the genesis and the maintenance of several human tumors, including neoplasms of the central nervous system (CNS). Growth factor autocrine and paracrine stimulatory loops promote tumor proliferation and angiogenesis. A family of structurally related growth factor receptors, the receptor tyrosine kinases, are particularly relevant to tumors of the CNS. This large family includes the receptors for the epidermal growth factor, the platelet-derived growth factor, the fibroblast growth factor, the insulin-like growth factor, the neurotrophins related to the nerve growth factor, and the vascular endothelial growth factor, as well as several receptors for which no growth factor ligand has been identified. Several of these receptor molecules and their growth factor ligands are preferentially expressed in the embryonic brain and are thought to play a central role in regulating the determination of the cell fate during the development of the CNS. Moreover, the overexpression or the mutation of genes encoding these receptors can be oncogenic. Researchers think that some receptors in this family (i.e., those that have been shown to be overexpressed or mutated in human brain tumors) contribute to brain tumor oncogenesis. This article will focus on recent experimental work and will discuss the classification and the biology of the receptor tyrosine kinases, as well as their roles in the development of the CNS and in tumorigenesis. PMID- 7477769 TI - Symptomatic cavernous malformations affecting the spine and spinal cord. AB - Ten cases of symptomatic cavernous malformations affecting the spine and spinal cord were retrospectively reviewed. The cases display a spectrum of pathological findings involving the vertebral body, vertebral body with epidural extension, epidural space without bony involvement, intradural extramedullary space, and intramedullary lesions. Lesions at all locations are identical histologically, electron microscopically, and immunohistochemically. This perspective, in which cavernous malformations are envisioned as a single entity arising at numerous locations, runs contrary to the view found in the neurosurgical literature. In most discussions of cavernous malformations, vertebral body lesions are depicted as separate entities from intradural lesions. Cavernous malformations, also called cavernous hemangiomas, are developmental vascular hamartomas that, by definition, do not grow by mitotic activity. Yet, the expansion of these lesions is well documented both in the literature and among our cases. The therapeutic modalities used in our series included observation, embolization, radiation, and surgical resection alone or in combination. All modalities are effective but must be tailored to the specific needs and condition of the patient. The embryology, methods of treatment, and proposed mechanisms of growth, plus similarities and differences between cavernous malformations at each location, are reviewed. Analogies between spinal and intracranial lesions are presented. On the basis of this series and a review of the literature, we conclude that cavernous malformations represent a single entity regardless of location. Segregation based on location, as is prevalent throughout the neurosurgical literature, hinders an overall understanding of these lesions. Cavernous malformations are more appropriately viewed as a single pathological entity arising in a multitude of locations. The difficulties encountered when managing cavernous malformations at various locations are unique to the location and not the lesion. PMID- 7477770 TI - Clinicopathological correlations in syringomyelia using axial magnetic resonance imaging. AB - Axial magnetic resonance (MR) images of non-neoplastic spinal cord cavities were reviewed in 115 patients with otherwise complete neurological and neuroradiological findings. The variations in axial morphology revealed three distinct cavitary patterns. These patterns were as follows: 1) symmetrically enlarged central cavities (28 patients); 2) central cavities that expanded paracentrally in one or more focal areas (36 patients); and 3) eccentric cavities that were off-center, frequently irregular, and sometimes associated with myelomalacia (51 patients). The radiological patterns of spinal cord cavitation correlated well with recently reported histopathological findings that distinguish simple dilations of the central canal, dilations of the central canal that dissect paracentrally, and primary cavitations of the spinal cord parenchyma (extracanalicular syringes). Like histologically confirmed central canal syringes, MR-defined central cavities were associated with pathogenic factors that affect the dynamics of the cerebrospinal fluid, including hindbrain malformations, hydrocephalus, and extramedullary obstructive lesions. Eccentric cavities resembled extracanalicular syringes and occurred typically with disorders that damage spinal cord tissue (e.g., trauma, infarction, meningitis/arachnoiditis, spondylosis/disc herniation, radiation necrosis, and transverse myelitis). Analysis of clinical findings at the time of MR imaging established the following correlations. 1) Symmetrically enlarged central cavities were asymptomatic or produced nonspecific neurological signs. 2) Central cavities that expanded paracentrally were associated with segmental signs referable to the paracentral component. 3) Eccentric cavities produced various combinations of long tract and segmental signs that could usually be related to the level, side, and specific quadrant of spinal cord cavitation.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7477771 TI - Posterior fossa volume and response to suboccipital decompression in patients with Chiari I malformation. AB - Smaller posterior fossa (PF) volume has been suggested to be one of the mechanisms responsible for tonsillar herniation through the foramen magnum in patients with Chiari I malformation (CM I). Although previous radiological analyses of the cranial anatomy have suggested a smaller PF volume in patients with CM I, the relationship of the PF volume to decompressive surgery has not been reported. We have measured the ratio of PF volume to supratentorial volume (PF ratio [PFR]) in 20 patients with CM I and 20 control patients by retrospectively studying their magnetic resonance images with a computerized image analyzer. The mean PFR in patients with CM I (with or without syringomyelia) was significantly smaller than for those in the control group (15.6 +/- 1.9 versus 17.5 +/- 1.2, P = 0.0008). Although PFR did not correlate with the extent of tonsillar herniation in patients with CM I, it did directly correlate with their age, i.e., younger patients with CM I (but not control patients) had smaller PFRs. All but three patients responded both clinically and radiographically to decompressive surgery. Those patients who did not benefit from surgical intervention had normal PFRs. We conclude that: 1) PFRs are smaller in most patients with CM I; 2) a smaller PF may be a primary cause of tonsillar herniation; 3) patients with CM I who have smaller PFRs tend to develop symptoms earlier than those with normal values; 4) patients with smaller PFRs tend to respond better to suboccipital decompression. PMID- 7477772 TI - Early discharge after carotid endarterectomy. AB - A consecutive series of 233 carotid endarterectomies performed over a 4-year period by one neurosurgeon was reviewed to analyze the number, nature, cause, and time to occurrence of complications. The time to discharge was analyzed regarding the type of anesthesia used and the time period during which surgery was performed. The records were reviewed to determine whether any complication could have been prevented by routine postoperative monitoring in an intensive care unit or by longer postoperative hospitalization. There were no perioperative deaths. The incidence of ipsilateral stroke was 1.7% and of major stroke was 0.9%. Nondisabling myocardial infarctions occurred in 1.3% of the patients. Complications occurred in a bimodal time course, less than 48 hours or greater than 1 week. The time to discharge decreased during the study period from 4.44 +/ 4.22 days for the first year of the study to 1.56 +/- 1.31 days for the last year of the study (P < 0.0001). A trend toward earlier discharge for patients who received cervical block anesthesia rather than general anesthesia did not reach statistical significance. No adverse patient outcome could be attributed to lack of intensive care unit monitoring or to early discharge. Our data demonstrate that patients who have undergone carotid endarterectomies do not routinely require intensive care unit monitoring and that discharge 24 to 48 hours after surgery is safe for the majority of patients. This regimen can result in considerable savings without compromising the quality of patient care. PMID- 7477774 TI - Hypertension and left ventricular hypertrophy in patients with spontaneous subarachnoid hemorrhage. AB - One hundred eighteen consecutive cases of spontaneous subarachnoid hemorrhage seen at one hospital during a 3-year period were examined to assess the prevalence of hypertension and the correlation between the presence of hypertension and the risk of early death. Eighty-seven of the patients had intracerebral aneurysms. The diagnosis of hypertension was determined by means of three complementary criteria: a history of treatment with antihypertensive drugs; systolic and/or diastolic blood pressure levels > or = 160 and 95 mm Hg, respectively, measured by the general practitioners of the patients before the onset of the subarachnoid hemorrhage; and the presence of left ventricular hypertrophy determined by echocardiography and/or necropsy. The major findings were as follows: 1) hypertension was present in at least 41% of the patients; 2) in 37% of 51 patients with no history of hypertension before the hemorrhage, left ventricular hypertrophy was diagnosed; and 3) the frequency of hypertension and left ventricular hypertrophy was significantly higher in patients who died within 14 days after the bleeding episode compared with patients surviving this period. PMID- 7477773 TI - Hemostasis in spontaneous subarachnoid hemorrhage. AB - To comprehensively examine changes in the hemostatic systems early after spontaneous subarachnoid hemorrhage (SAH) and to assess the relationships between those changes and neurological findings, computed tomographic findings, and clinical outcomes, we reviewed 167 patients who were admitted within 24 hours of the onset of SAH. Blood was taken from all of the patients at admission for detailed examinations of the hemostatic systems. The following results were obtained: 1) the levels of the thrombin-antithrombin complex (elevation indicating the activation fo the blood coagulation system), plasmin-antiplasmin complex, and D-dimer (elevation indicating the activation of the fibrinolytic system) significantly increased with the neurological severity, amount of subarachnoid clot, and severity of clinical outcome; 2) the levels of the thrombin-antithrombin complexes and plasmin-antiplasmin complexes were significantly higher in patients with intracerebral or intraventricular hematomas than in patients without those hematomas; 3) in each of almost all the neurological and computed tomographic grades, the levels of the thrombin antithrombin complexes were significantly higher in the patients with poor outcomes than in those with good outcomes; 4) the poor outcome rate significantly increased with neurological severity and the amount of subarachnoid clot. These data suggest that the activation of the blood coagulation system as well as of the fibrinolytic system occurred early in the course of SAH and that these systems were activated to a greater extent with the severity of neurological grades, computed tomographic findings, and clinical outcome.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7477775 TI - Elevated levels of lipoprotein (a) in association with cerebrovascular saccular aneurysmal disease. AB - The relationship between cerebrovascular aneurysmal disease and atherosclerosis remains unclear. Elevated serum levels of lipoprotein (a) (Lp[a]), are an independent risk factor for atherosclerosis. We measured serum Lp(a) levels in 50 patients who had angiographically proven saccular aneurysmal disease and who were free of clinically significant atheromatous disease (as judged by their medical histories and the results of physical examination, electrocardiography, and carotid angiography). The Lp(a) serum levels in these patients were compared with the Lp(a) serum levels in a group of 42 normal healthy controls. Serum Lp(a) levels in the patients was 20.1 +/- 0.42 mg/dl (median +/- standard error); however, median serum Lp(a) in the control subjects was 10.8 +/- 0.47 mg/dl (P < 0.01). Among females, the difference in serum Lp(a) levels was significant; the levels were 22.2 +/- 0.6 for female patients (n = 29) and 9.5 +/- 0.53 in female control subjects (n = 26) (P < 0.005). The most significant difference (P < 0.002) was seen in females < 50 years old (14 patients, 10 control subjects). No significant differences were seen in the Lp(a) serum levels between 21 male patients and 16 male control subjects. Lp(a) levels above the threshold level (30 mg/dl) were found in 20 patients and 7 control subjects (chi 2 = 5.99, P < 0.02); 12 female patients and 3 female control subjects (chi 2 = 6.16, P < 0.02; 8 male patients and 4 male control subjects (this difference was not significant). These results indicate either that cerebrovascular aneurysmal disease and subclinical atherosclerosis are related or that Lp(a) is a risk factor for vasculopathies other than atheroma. PMID- 7477776 TI - Aberrant p53 expression does not correlate with the prognosis in anaplastic astrocytoma. AB - Mutations of the p53 tumor-suppressor gene, as determined by the immunohistochemistry of archival formalin-fixed specimens, have been correlated with the prognosis for patients with many different types of malignancy. Similar correlations have been shown in series including patients with all grades of astrocytoma. We hypothesized that this might be a clinically useful prognostic indicator for patients with a defined grade of astrocytoma, anaplastic astrocytoma. This series comprised 54 consecutive patients with biopsy-proven anaplastic astrocytoma treated at one institution. When the CM-1 antiserum was used for testing, 33 (61%) of the 54 biopsies exhibited positive nuclear staining for p53, indicating an abnormal accumulation of the protein. This staining was graded according to the number of positive cells per high-power field. Positive immunohistochemical staining for p53 in the tumor cell nuclei did not correlate with the patient's outcome. Significant correlates of improved patient survival were the presentation with epilepsy in the absence of a neurological deficit (P = 0.005) and the surgeon's performance of a macroscopically complete surgical resection of the tumor (P = 0.01). PMID- 7477777 TI - The mapping and continuous monitoring of the intrinsic motor nuclei during brain stem surgery. AB - A comprehensive technique was developed for continuous electrophysiological monitoring of intrinsic brain stem motor function during surgery to remove space occupying lesions in the fourth ventricle and brain stem. The technique is analogous to that used during surgery in the cerebellopontine angle; motor nuclei and peripheral pontine fiber tracts of Cranial Nerves III-XII are identified by the electrical stimulation of structures in the operative field and the evaluation of the compound muscle action potentials recorded from the corresponding muscles of the head. Nerve function is monitored continuously by recording the ongoing electromyographic activity in these same muscles. Broadcasting electromyographic responses through a loudspeaker gives the surgeon immediate feedback on the status of the motor nuclei being monitored. Advantages of this technique include 1) the positive, objective identification of the nuclei and fiber tracts; 2) the continuous feedback on the status of these structures; 3) a safe approach through the fourth ventricle to the lesions in the brain stem; 4) the positive identification of the boundaries between the neoplasm and the motor structures of the rhomboid fossa; and 5) a warning to the surgeon of potentially harmful nerve manipulations (contact, dissection, transection) during surgery. After this technique was used in 16 consecutive operations to remove cavernomas (n = 9), gliomas (n = 4), and other types of tumors (n = 3), surgical and neurological results showed the method to be reliable and simple to perform. PMID- 7477778 TI - Preemptive spinal cord stimulation reduces ischemia in an animal model of vasospasm. AB - Spinal cord stimulation (SCS) has been increasingly used in the treatment of ischemia caused by obliterative diseases in the extremities and in the cardiac circulation. The most promising effects have been obtained when physicians suspect that a major vasospastic component underlies the ischemic symptoms (e.g., as in Raynaud's disease). Despite the clinical success of this treatment method, little is known about the mechanisms underlying the pain relief it produces and its anti-ischemic effects. Most earlier experimental studies have used normal animals or animals with cerebral vasospasm induced by injection of autologal blood into the cerebrospinal fluid space. In the present study, we applied SCS in a rat model via implanted electrodes to study the effect of preemptive stimulation on the ischemia caused by vasospasm in a neurovascular flap in the groin; the vasospasm was induced by mechanical pressure applied to the feeding artery. In rats treated with SCS, delivered with parameters similar to those used clinically, the percentage of flaps recovering normal microcirculation after the spasm was significantly higher than in the untreated control group (100 and 28%, respectively; P < 0.05), and the maximal blood flow after the ischemic episode was significantly higher in the SCS group than in the control group (127 and 51 arbitrary units, respectively; P < 0.05). The percentage of animals regaining the premanipulation circulation after the provocation of a second spasm was also greater in the SCS-treated group than in the control animals (50 and 14%, respectively; P < 0.05). Pilot studies showed that this protective effect was specific to SCS given before spasm induction, an observation corroborated by clinical experiences.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7477779 TI - Treatment of head injury in mice, using a fructose 1,6-diphosphate and dimethyl sulfoxide combination. AB - Mice were subjected to moderate (800 g/cm force) or severe (900 g/cm force) head injury and treated 5 minutes later with various compounds. Treatments consisted of intravenous administration of the following compounds: 1) fructose 1,6 diphosphate (FDP), 2) dimethyl sulfoxide (DMSO), 3) FDP + DMSO, and 4) vehicle nontreated. Sensory-motor evaluations 1 and 2 hours after 800 g/cm-force head injury showed that significant protection of motor function (grip test) was achieved with FDP + DMSO but not with treatment by either drug alone. Evaluation of mice following a severe 900-g/cm force injury demonstrated significant survival after treatment with FDP + DMSO but not with the vehicle or treatment by either of these agents alone. Histopathological morphometry indicated that cortical and hippocampal CA1 neurons were markedly protected from damage when mice were treated with combined FDP + DMSO. More modest protection of CA1 but not of cortical neurons was observed after treatment with DMSO alone but not after treatment with FDP alone or administration of the vehicle. These findings indicate that combining FDP with DMSO results in considerable synergy in protecting animals from sensory-motor loss and neuronal brain damage and in ultimate survival stemming from a moderate or severe closed head injury. PMID- 7477780 TI - Use of local cerebral blood flow monitoring to predict brain damage after disturbance to the venous circulation: cortical vein occlusion model by photochemical dye. AB - A rat model of cortical vein occlusion by the photochemical thrombotic technique was used to evaluate whether monitoring the change in the local cerebral blood flow (LCBF) could predict brain damage after cortical vein occlusion. The cortical vein occlusion was attained by using the photochemical thrombotic technique, and the sequential LCBF was measured by using a laser Doppler flowmeter positioned over the middle frontal cortex between two adjacent dorsal veins for 120 minutes after the start of the irradiation to the cortical vein. Rats were assigned to one of three experimental groups. In Group A (n = 10), one dorsal cerebral vein was occluded; in Group B (n = 10), two adjacent dorsal cerebral veins were occluded; and in a sham-operated group (n = 5), the rats also underwent craniotomy and light exposure but received injections of saline rather than the rose bengal dye. After 24 hours, the rats were submitted to perfusion fixation and were examined histopathologically. After irradiation-induced cortical vein occlusion, a gradual and significant decrease of the LCBF was observed in both Group A (significantly different from the LCBF of the sham operated group after 90 min, P < 0.05, and 120 min, P < 0.01) and Group B (significantly different from the LCBF of the sham-operated group at 30 min, P < 0.05, and at 60, 90, and 120 min, P < 0.01). Significant differences (P < 0.01) in the LCBF were also seen between the two experimental groups at 90 and 120 minutes after the start of the irradiation and the subsequent brain damage.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7477781 TI - Copper chelation inhibits tumor angiogenesis in the experimental 9L gliosarcoma model. AB - To investigate the effects of copper (Cu)-depletion diet and D-penicillamine treatment (CDPT) on both tumor growth and angiogenesis, we studied Fischer-344 rats in which 9L gliosarcoma cells had been subcutaneously implanted. We focused primarily on the alteration of Cu contents and the vascular density. Compared with the normal diet group, the CDPT group showed a significant reduction of tumor weight and a decrease in Cu concentration. Furthermore, the CDPT group demonstrated smaller blood vessels with significantly lower vascular density. This decrease of tumor growth was achieved by angiosuppression. Our study indicated that CDPT selectively caused Cu chelation from the tumor tissue; the normal brain tissue did not show lower Cu concentration after the treatment. The prevention of tumor angiogenesis by this method may be very useful in cancer therapy and may help elucidate the microenvironmental mechanisms for cancer cells. PMID- 7477782 TI - Substrates for astrocytoma invasion. AB - A better understanding of the influences of specific extracellular substrates, including proteins, glycosaminoglycans, and parenchymal cells, on the invasive behavior of glioma cells would potentially lead to novel forms of treatment aimed at confining the tumor. A monolayer, microliter scale assay was used to investigate how different substrates influenced glioma migration. Basal or unspecific movement (range, 10-260 microns/d) was determined by observing a panel of seven established human glioma cell lines. Migration rates two to five times higher than this basal activity were referred to as preferential and specific glioma migration; these rates generally occurred on merosin and tenascin. Collagen, fibronectin, or vitronectin were less supportive of migration. The glioma cells migrated on hyaluronic acid, but they did not migrate to the extent generally found on the extracellular matrix proteins. Glioma-derived extracellular matrix also served to promote cell migration. This finding implicates a role for either glioma remodeling or synthesis of a permissive environment for local dissemination that may be independent of the constitutive matrix proteins normally found in the brain. Although the glioma cells were able to migrate over monolayers of other glioma cells, they were unable to migrate over astrocytes and fibroblasts. Our findings indicate that the invasive behavior of glioma cells in situ is most likely a consequence of the interplay between the cells' manipulation of the environment and the constitutive ligands associated with specific regions or structures of the brain. PMID- 7477783 TI - Anatomy of the thoracic pedicle. AB - Thoracic pedicle anatomy (interpedicular distance, transverse and sagittal pedicle widths, transverse and sagittal pedicle angles, and the distance from the axis of the pedicle to the axis of the transverse process) was assessed in 11 cadavers of elderly people. The cadaveric spines were extensively dissected to augment the accuracy of the measurements via caliper and goniometer. The results were compared with those of previous studies that assessed pedicle anatomy with computed tomography, direct measurement, and three-dimensional morphometry. Between the studies, significant differences were found in transverse pedicle width and transverse and sagittal pedicle angles. These morphometric differences may reflect either the diversity of the techniques used to measure the pedicle anatomy or sampling variation. This article presents a previously unreported morphometric finding, the rostral-caudal distance from the thoracic pedicle to the midpoint of the base of the transverse process. At T1, the transverse process is 5.45 +/- 1.2 mm rostral to the pedicle. This relationship gradually changes as the thoracic spine is descended, so that at T12, the transverse process is 6.6 +/ 2.4 mm caudal to the pedicle. Crossover consistently occurs at the T6-T7 region. Although the transverse process is a reliable external landmark for the location of the pedicle in the lumbar spine, this relationship in the thoracic spine is variable and only moderately predictable. PMID- 7477784 TI - The blood supply of the trigeminal nerve root, with special reference to the trigeminocerebellar artery. AB - The vasculature of the 29 roots of the trigeminal nerve was examined after india ink and gelatin had been injected into the vertebrobasilar arterial system. The trigeminal arteries were most often noted to arise from the superolateral pontine branch of the basilar artery (89.66%), and from the peduncular cerebellar branch of the anterior inferior cerebellar artery (75.86%). The trigeminocerebellar artery supplied two roots (6.89%) of the trigeminal nerves. The number of trigeminal arteries ranged from two to six, and their diameters ranged from 100 to 510 microns. Anastomoses among them were seen in 37.93% of the cases. The arteries formed the vascular rings around 58.61% of the roots. The motor portion of the trigeminal nerve most often received blood from the superolateral pontine artery (79.31%). The same artery most commonly supplied the rostral part of the sensory portion, which corresponded to the ophthalmic division of the trigeminal nerve. The superolateral artery, together with the inferolateral pontine artery and the peduncular cerebellar branch of the anterior inferior cerebellar artery, irrigated the middle part of the sensory portion, which corresponded to the maxillary division. The caudal part of that portion, which corresponded to the mandibular division, was commonly perfused by the peduncular cerebellar branch of the anterior inferior cerebellar artery. In this article, we discuss the possible clinical significance of the anatomic data observed. PMID- 7477785 TI - Resolution of chronic cluster headache after resection of a tentorial meningioma: case report. AB - Cluster headache is almost always idiopathic, but, in rare cases, associated intracranial lesions have been found. We describe a patient who had chronic cluster headache for more than 20 years. The headache immediately resolved upon resection of a tentorial meningioma. Prior reports of cluster headache as a manifestation of structural disease are briefly reviewed. In the patient described, the pain was referred from the right tentorium cerebelli to the right side of the face, in accordance with reported studies on the subjective localization of pain referred from posterior fossa structures. The accompanying abnormalities of autonomic function may have been mediated by central autonomic reflexes that are also involved in the pathogenesis of idiopathic cluster headache. PMID- 7477786 TI - Concurrent trigeminal, abducens, and facial nerve palsies presenting as false localizing signs: case report. AB - Multiple cranial nerve dysfunction presenting as false localizing signs is rare. We report a 20-year-old woman who had concurrent trigeminal sensory disturbance, abducens, and peripheral facial nerve palsies in association with a contralateral acoustic neuroma. After surgery, the trigeminal nerve disturbance and the abducens nerve palsy completely recovered, but the peripheral facial nerve palsy persisted. The nature of tumor, the presence of brain stem distortion, the anatomic variation of posterior fossa, and the relationships of cranial nerves and nearby blood vessels, which are factors influencing the occurrence of false localizing, are briefly discussed. PMID- 7477787 TI - Multiple intracerebral metastases from an islet cell carcinoma of the pancreas: case report. AB - Computed tomography and magnetic resonance imaging of an 85-year-old woman with bitemporal headaches revealed cystic lesions in the basal ganglia and cerebellum. The patient remained neurologically asymptomatic until just before her death at age 89. Autopsy revealed tumors of the brain, lungs, liver, periaortic lymph nodes, adrenal gland, and the pancreas. Light microscopy demonstrated a malignant small cell tumor with scant cytoplasm. Electron microscopy revealed intracytoplasmic dense core vesicles, characteristic of neuroendocrine tumors, which prompted the diagnosis of islet cell carcinoma. All lesions were found to be microscopically similar. Previous reports of islet cell carcinoma metastatic to the brain could not be found. This case emphasizes the importance of electron microscopy in cases of small cell neoplasms in the brain. PMID- 7477788 TI - Primary intracranial squamous cell carcinomas: report of two cases. AB - Two cases of primary intracranial squamous cell carcinomas are reported. The first patient is a 57-year-old man with an epidermoid carcinoma in the right cerebellopontine angle, having histological features that indicated malignant transformation in a benign epidermoid cyst. The second patient is a 42-year-old man in whom squamous cell carcinoma arose in a pre-existing middle fossa dermoid cyst 9 years after the initial surgical resection. The literature on primary intracranial squamous cell carcinomas is reviewed, and the clinical features and histological pathogenesis for the occurrence of this unique condition are discussed. PMID- 7477789 TI - Intraneural neurothekeoma: case report. AB - Neurothekeomas are rare, benign cutaneous tumors, with a distinctive histological appearance, that seldom have been documented to arise from identifiable medium- or large-caliber peripheral nerves. We report a neurothekeoma of the left posterior tibial nerve, whose gross histological appearance and strong immunostaining for S-100 in the majority of the tumor corroborates the impression that this tumor is a variant of a schwannoma. PMID- 7477791 TI - Spontaneous regeneration of the foramen magnum after decompressive suboccipital craniectomy in Chiari malformation: case report. AB - A 21-year-old woman, who had undergone foramen magnum decompression for her symptomatic Chiari malformation (Type 1) 7 years before, presented with recurrence of the symptoms. Neuroradiological examinations demonstrated regeneration of the foramen magnum that was caused by new bone formation, as confirmed at the second surgery. Neurological improvement was obtained after the removal of the regenerated foramen magnum. This observation, though rare, deserves to be kept in mind during the postoperative follow-up period in young patients with Chiari malformation. PMID- 7477790 TI - Tumoral calcium pyrophosphate dihydrate deposition disease mimicking a cervical spine neoplasm: case report. AB - A case of tumoral calcium pyrophosphate dihydrate crystal deposition disease involving the upper cervical spine is reported. It presented clinically, radiographically, and by preliminary intraoperative pathological evaluation as a possible malignant soft tissue tumor. An aggressive resection of the lesion was performed. This case differs from previous reports of calcium pyrophosphate dihydrate crystal deposition disease of the cervical spine by the size, location, and radiographic appearance of the lesion. We suggest that radical surgical procedures should not be performed without consideration of this diagnosis in lesions with similar presentations. A brief review of spinal and tumoral calcium pyrophosphate dihydrate crystal deposition is presented. PMID- 7477792 TI - Central lumbar stenosis caused by ligamentum flavum: unilateral laminotomy for bilateral ligamentectomy: preliminary report of two cases. AB - Degenerative central lumbar stenosis has traditionally been considered to be a result of bony narrowing of the spinal canal. In two consecutive patients with degenerative central lumbar stenosis and complete myelographic blocks, the cauda equina was compressed by a thickened ligamentum flavum (cross-sectional area [CSA], > 150 mm2). This ligamentous stenosis occurred within bony canals of normal dimensions (anteroposterior diameter, interpediculate distance, interfacet distance, and CSA). High-resolution computed tomographic myelography was used to calculate quantitative values for the CSA, the length, and the volume of the lumbar ligamentum flavum. Excision of the thickened ligamentum flavum restored the dural sac to normal (CSA, 130-230 mm2), and both patients received relief from their symptoms. Unilateral laminotomy was used successfully to achieve bilateral ligamentectomy. Therefore, in a well-defined subgroup of patients with degenerative central lumbar stenosis, the dural sac can be decompressed by selective resection of the ligamentum flavum, and bilateral ligamentectomy can be performed via unilateral laminotomy. PMID- 7477793 TI - William Macewen (1848-1924). PMID- 7477794 TI - Perspectives in neurosurgery: neurosurgery in Greece. PMID- 7477795 TI - Management of 100 consecutive direct carotid-cavernous fistulas: results of treatment with detachable balloons. PMID- 7477796 TI - The avoidance of surgery in the treatment of subarachnoid cutaneous fistula by the use of an epidural blood patch: technical case report. PMID- 7477797 TI - Idiopathic chronic hypertrophic craniocervical pachymeningitis: case report. PMID- 7477798 TI - Propylene glycol toxicity following continuous etomidate infusion for the control of refractory cerebral edema. AB - Continued elevations in Intracranial Pressure (ICP) following traumatic or ischemic compromise are known to cause markedly increased morbidity and mortality. Because of the side effects of barbiturates including hypotension and prolonged recovery time, the use of shorter-acting anesthetic agents to control ICP has been considered. Etomidate, when administered by continuous infusion, has been shown to decrease cerebral metabolism resulting in a secondary decrease in cerebral blood flow with minimal changes in cerebral perfusion pressure. We initially intended to randomize 20 patients prospectively into a study protocol that would assess the effects of either pentobarbital or the cardioprotective agent etomidate on ICP and cardiac performance. Given the sequelae of the therapy, we were only able to randomize seven patients with cerebral edema refractory to medical management to receive either etomidate or pentobarbital in a blinded fashion. Three patients who received etomidate developed renal compromise (mean low creatinine clearance 41 ml/min, range 37-44 ml/min) which was initially noted at 24 hours. We believed that this represented an adverse effect that was probably related to the study drug and the study was stopped. Each patient received a 0.30 mg/kg IV induction of etomidate and then 0.02 mg/kg/min continuous infusion for 24-72 hours titrated burst suppression. All patients also received dexamethasone 2 mg IV every six hours to prevent the adrenocortical insufficiency that might occur as a consequence of etomidate induced suppression of cortisol synthesis. Intracranial pressure decreased (mean = 12mmHg) following the initiation of etomidate. Cardiac parameters remained unchanged (cardiac output 4.8 +/- .6 liters/min).(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7477799 TI - On the choice of recording duration in pharmaco-EEG studies. AB - Quantitative EEG is a sensitive method used to assess the effects of pharmacological substances on the central nervous system (CNS) activity. A standard technique is to measure the EEG under vigilance-controlled and resting conditions for a short duration, for example 5 min. The aim of the present study was to investigate the stability of 5-min EEG recordings. While the time course of the EEG was fairly stable during the recording session under the vigilance controlled condition, systematic trends became apparent under the resting condition. Pharmaco-sensitivity of the EEG and its reliability increased with the recording duration. Five minutes of EEG recording seem to be sufficient and well chosen to evaluate the influence of drugs on the EEG. PMID- 7477800 TI - Combined cognitive-behavioral, psychopharmacological and nutritional therapy in eating disorders. 1. Anorexia nervosa--restricted type. AB - Twenty-two female patients with anorexia nervosa, restricted type, 14-35 years old, were treated with a 4-month course of combined cognitive-behavioral therapy, nutritional counselling and antidepressant drugs (nortriptyline for 7, fluoxetine for 15). Patients were monitored for body mass index (BMI), for eating disorder symptoms by the Eating Disorder Inventory (EDI) and the Bulimic Investigation Test (BITE) and for depression and anxiety by the Hamilton Rating Scales for Depression and for Anxiety (HRS-D and -A). The scores were determined before and after 1, 2 and 4 months of therapy. BMI, depression, anxiety and EDI scores improved significantly and equally in both groups during the 4 months of therapy, while BITE scores did not change. PMID- 7477801 TI - Combined cognitive-behavioral, psychopharmacological and nutritional therapy in eating disorders. 2. Anorexia nervosa--binge-eating/purging type. AB - Thirteen women with anorexia nervosa, binge-eating/purging type (AN-BP), 17-43 years old, were treated with a 4-month course of combined cognitive-behavioral, nutritional and antidepressant therapy (7 with amineptine and 6 with fluoxetine). Patients were monitored before and after 1, 2 and 4 months of treatment for body mass index (BMI), for eating disorder symptoms by the Eating Disorder Inventory (EDI) and the Bulimic Investigation Test (BITE) and for depression and anxiety by the Hamilton Rating Scales for Depression and for Anxiety. BMI, EDI scores, depression and anxiety improved significantly and equally in the two groups during the 4 months of therapy, while BITE scores did not change. PMID- 7477802 TI - Combined cognitive-behavioral, psychopharmacological and nutritional therapy in bulimia nervosa. AB - Fifteen women with bulimia nervosa were treated with a 4-month course of combined cognitive-behavioral, nutritional and antidepressant therapy (5 with amineptine and 10 with fluvoxamine). Patients were monitored before and after 1, 2 and 4 months of therapy for body mass index (BMI), for eating disorder symptoms by the Eating Disorder Inventory (EDI) and the Bulimic Investigation Test (BITE), and for depression and anxiety by the Hamilton Rating Scale for Depression and for Anxiety (HRS-D and -A). BITE symptoms and gravity improved significantly and equally in the two groups during the 4 months of therapy. Global EDI scores, depression and anxiety decreased but not significantly. BMI was normal before therapy and did not change during treatment. PMID- 7477803 TI - P300 in posttraumatic stress disorder. AB - In the present study, P300 has been recorded in 26 subjects (15 women) 1 month after an aggression without organic complications. Among our sample, 16 subjects fulfilled DSM-III-R criteria for posttraumatic stress disorder (PTSD) and 10 did not. P300 amplitude was significantly lower in the 16 PTSD subjects as compared to the 10 subjects without PTSD. This study supports information processing disturbances in PTSD. PMID- 7477804 TI - The association of the serotonin metabolite, 5-HIAA, to the lysosomal enzyme N acetyl-beta-glucosaminidase. AB - Twenty-one medication-free patients with generalized anxiety disorder (GAD) collected urine samples for 5-hydroxyindoleacetic acid (5-HIAA) and N-acetyl-beta glucosaminidase (NAG). Nine patients with no comorbid psychiatric diagnosis showed a significant association between 5-HIAA and NAG, r = -0.683, p = 0.04. The possibility that NAG could be a marker for serotonin metabolism is discussed. PMID- 7477805 TI - A historical case of disseminated chronic tuberculosis. AB - A British author has put forward a new explanation for Joan of Arc's behaviour. This author suggests that the voices Joan of Arc heard were due to the presence of a temporal lobe tuberculoma in the context of widespread chronic tuberculosis (exposure to bovine tuberculosis, amenorrhoea, heart and intestines incombustible). We consider that some elements are incompatible with widespread tuberculosis. It is difficult to draw final conclusions, but it would seem unlikely that widespread tuberculosis, a serious disease, was present in this 'patient' whose life-style and activities would surely have been impossible had such a serious disease been present. PMID- 7477806 TI - Pathophysiology and psychopharmacology of dementia--a new study design. I. Diagnosis comprising subjective and objective criteria. AB - In a double-blind placebo-controlled psychopharmacological study 263 patients with cognitive impairment were screened and 137 of them were selected (following defined inclusion and exclusion criteria) to participate in a clinical trial (for results of treatment phase see part II, in preparation). The pretreatment state was evaluated twice using quantitative-topological EEG (qEEG) and psychopathological rating scales. There is a significant correlation between total score values of nearly all dementia-specific neuropsychological rating scales and absolute spectral power in the lower frequency ranges (delta, theta, alpha 1) at rest. In order to increase the diagnostic power, qEEG was recorded simultaneously with the performance in four different psychometric tests. It was found that the worse the psychopathological rating of a patient, the higher the absolute theta power at rest and the smaller the test- and demand-dependent further increase in theta power. The percentage of task-induced power increase in delta and theta frequency range appeared to be an efficient indicator for a reliable discrimination between healthy elderly individuals and demented patients (rank correlation coefficient for theta in F7: r = +0.55), which will also serve for evaluation of treatment. PMID- 7477807 TI - Vitamin supplementation for 1 year improves mood. AB - The possibility that the taking of vitamin supplements may influence mood was explored. One hundred and twenty-nine young healthy adults took either 10 times the recommended daily dose of 9 vitamins, or a placebo, under a double-blind procedure, for a year. Males taking the vitamins differed from those taking the placebo in that they reported themselves as feeling more 'agreeable' after 12 months. After 12 months the mood of females taking the vitamin supplement was significantly improved in that they felt more 'agreeable', more composed and reported better mental health. These changes in mood after a year occurred even though the blood status of 9 vitamins reached a plateau after 3 months: this improvement in mood was associated in particular with improved riboflavin and pyridoxine status. In females baseline thiamin status was associated with poor mood and an improvement in thiamin status after 3 months was associated with improved mood. PMID- 7477808 TI - Sustained arousal and attention after traumatic brain injury. AB - Clinicians report that patients with traumatic brain injury (TBI) often have difficulty with tasks requiring sustained attention, and there are neuroanatomical and neurophysiological reasons to expect such deficits. Nevertheless, laboratory measures of sustained attention or vigilance in TBI have produced conflicting results. These inconsistencies may be due to patient heterogeneity as well as the fact that vigilance performance is dependent on highly specific features of the task design. We developed a visual vigilance task in which the influence of non-attentional factors was minimized and task difficulty for patients and controls made comparable. Performance was characterized with respect to vigilance level as well as vigilance decrement, using measures of perceptual discrimination, response bias, reaction time and reaction time variability. Twenty-six patients with recent TBI and 18 control subjects were tested on this task. A MANOVA of ranked scores revealed significantly different patient and control performance overall. Initial level of performance (vigilance level) was slower and more variable for patients than controls, and patients showed more conservative response biases. Deterioration over time (vigilance decrement) was steeper for patients than controls for reaction time, reaction time variability, and response bias. Deterioration in accuracy (D') did not differ significantly between patients and controls. Performance was not related to available measures of injury severity. Hypotheses relating arousal mechanisms to vigilance performance are discussed. PMID- 7477809 TI - Which hemisphere falls asleep first? AB - Behavioral tasks (reaction times to acoustic stimuli and finger tapping tasks) performed by normal subjects when sleepy or attempting to fall asleep have been used as indices of hemispheric asymmetries during the sleep onset period. Results show a stronger impairment of the left hemisphere (right hand) both in reacting to external stimuli and in sustaining endogenous motor programs. The left hemisphere seems to fall asleep earlier than the right hemisphere. PMID- 7477810 TI - Study of unilateral hemisphere performance in children with developmental dysphasia. AB - Hemisphere specialization for language was studied in 10 children with expressive developmental dysphasia (DD) (mean age 10 years 4 months) submitted to a dichotic listening task (in a word free-recall task and forced-attention task) and a finger tapping/vocalization dual-task paradigm. A nonsense shape dichaptic task was also introduced to control right hemispheric processing. Performances of dysphasic children were compared to those obtained from 15 normal children. The results showed that controls had a right ear advantage in free-recall (words) dichotic listening task and a significant right ear advantage in forced-right attention task, with a change in ear asymmetry as a consequence of instruction. In the dysphasic group we observed a significant right ear advantage in the free recall dichotic listening task and no change in ear asymmetry during forced right or forced left condition. Results in time sharing paradigm and nonsense dichaptic task are more difficult to interpret, because there was no interaction between group and condition. These results cannot support a complete left hemisphere dysfunction in developmental dysphasia. PMID- 7477811 TI - Hemisphere asymmetry and the processing of emotional words in anxiety. AB - High- and low-trait anxious subjects participated in a divided visual field emotional Stroop task. Subjects identified the colour of unilaterally presented threat-related, positive and neutral words. The high- and low-trait groups differed in terms of their accuracy scores but not in their reaction times. Analysis of the accuracy data revealed that the high-trait group exhibited right hemisphere interference for emotional relative to neutral trials whereas the low trait showed right hemisphere interference for threat-related trials only. The idea that the right hemisphere is differentially specialised for the processing of emotional information was supported. PMID- 7477812 TI - Blood glucose levels selectively influence memory for word lists dichotically presented to the right ear. AB - The relationship between performance on a dichotic listening task and blood glucose levels was examined. It was predicted that, if the ability of blood glucose to supply the brain with its basic fuel limits performance under conditions of cognitive demand, in a dichotic listening task information directed to the left hemisphere would be particularly susceptible to the level of blood glucose. Via headphones subjects heard lists of words directed to both ears, although randomly they were asked to attend to those directed to one ear. Those receiving a glucose drink recalled more of a list of words directed to the right ear and hence the left hemisphere. Subjects with low baseline blood glucose levels recalled more from the attended ear, and those with high baseline blood glucose more from the unattended ear. In those who received a glucose drink, a fail in blood glucose during the dichotic task was associated with a right ear superiority. The pattern of findings can be explained by the assumptions that under conditions of cognitive demand the supply of glucose to the brain limits performance and that there are individual differences in the ability to efficiently take glucose from the blood stream into the brain. PMID- 7477814 TI - Healthy aging, memory subsystems and regional cerebral oxygen consumption. AB - The present study was designed to search for concomitant age-related changes in memory subsystems, defined according to current structural theories, and resting oxygen consumption in selected brain regions. We have investigated a sample of subjects between 20 and 68 years of age and strictly screened for their good health. We applied in the same subjects a battery of neuropsychological tests selected to investigate several memory subsystems, and high-resolution positron imaging with stereotaxic localization to study a purposely limited number of cerebral structures, selected on a priori hypotheses to match the different memory subsystems. Our results showed significant age-related changes in performance on some tests, consistent with the literature, including an increase in semantic memory and a decrease in both working memory (central executive system) and verbal episodic and explicit memory. There was also an age-related linear decrease in global brain oxygen consumption which regionally reached statistical significance for the neocortical areas and the left thalamus. There was a limited number of significant, age-independent correlations between the raw psychometric test scores and resting regional oxidative metabolism. Consistent with our present understanding of the functional anatomy of memory, the Associate Learning scores (verbal episodic and explicit memory) were positively correlated with left hippocampal and thalamic metabolism. The positive relationships found between right hippocampal metabolism and performance in the Associate Learning and the Brown-Peterson tests were less expected but would be consistent with findings from recent PET activation studies. The results from this investigation are discussed in the light of current knowledge concerning the neuropsychology and the neurobiology of both aging and memory. PMID- 7477813 TI - Menstrual cycle affects functional cerebral asymmetries. AB - Cerebral lateralization in females is probably modulated by the menstrual cycle such that a part of hemispheric asymmetries are diminished with an increase of the steroids estrogen and progesterone during the follicular and luteal phase and enhanced with steroid decreases during the menstrual phase. However, previous data were contradictory with regard to the hemispheric side of modulation and could not analyze which steroid is mainly responsible for cycle dependent lateralization shifts. Therefore, in the present study estrogen and progesterone changes were assessed separately and related to changes in cerebral asymmetry. Plasma levels of estrogen and progesterone were measured once during luteal and once during menstrual cycle phase while 20 females subjects performed a verbal and a figural lateralized matching task. The results showed a significant cycle phase x lateralization interaction for the right hemisphere dominated figural comparison task but not for the left hemispheric lexical condition. Although the lateralization was modulated by the menstrual cycle, a within-subject regression analysis demonstrated that the asymmetry shift was not under direct influence of estrogen or progesterone. Thus, the present study provides further empirical support for cycle dependent alterations in lateralization but makes it unlikely that this effect is directly caused by estrogen or progesterone plasma level variations. PMID- 7477815 TI - Electrophysiological signs of sustained and transient attention to spatial locations. AB - Event-related potentials were elicited by bilateral and unilateral stimulus arrays flashed in rapid sequence in order to investigate both focused attention and attentional orienting. Subjects attended selectively to the stimuli on one side of the bilateral arrays and were required to discriminate infrequent target stimuli on either the attended side (no switch of attentional focus) or unattended side of the array (switch of attentional focus). The ERPs to the bilateral stimuli elicited an occipital P1 component that was larger in amplitude over scalp regions contralateral to the attended visual half-field. The ERPs to the unilateral stimuli on the attended side also showed an amplitude enhancement of early P1 components, followed by a positive shift that lasted until 200 msec latency over the contralateral occipital scalp. No enhancement of the N1 component was observed to attended-side stimuli. These patterns were not different for conditions requiring or not requiring a spatial switch of the attentional focus. In conjunction with ERP signs of focused spatial attention, significant differences in discrimination performance (d') were obtained for the attended vs unattended-side targets; no changes in measures of criterion (beta) were obtained. These data support the idea that the early occipital P1 attention effect represents a facilitation of visual inputs that occur at attended locations in the visual field. PMID- 7477817 TI - Lateralized dual task interference in left-handers: initial value differences do not affect the outcome. AB - The study was conducted to clarify factors involved in dual-task finger-tapping interference. Left-handers, as assessed by hand-writing preference and left-hand baseline tapping advantage, tapped both alone and while solving anagrams. Even though the left-hand baseline tapping advantage was experimentally removed on some (adjusted) trials, greater left- than right-hand tapping interference was observed during concurrent task performance. This result coupled with previous findings for right-handed subjects [Kee and Cherry, Neuropsychologia, Vol. 28, pp. 313-316, 1990] indicates that lateralized interference effects are not merely due to initial baseline tapping differences as proposed by Willis and Goodwin [Neuropsychologia, Vol. 25, pp. 719-724, 1987]. PMID- 7477816 TI - A dissociation between developmental surface and phonological dyslexia in two undergraduate students. AB - This study compares the nature of the reading deficit that was observed in two dyslexic undergraduate students who were severely impaired at reading and spelling compared with normal undergraduates. They both achieved the same (below average) score on the National Adult Reading Test and on the Schonell spelling test. One of them, however, was good at reading and spelling nonwords, had good phonological awareness skills, was better at reading regular than irregular words, and made phonologically accurate reading and spelling errors (i.e. was a surface dyslexic). The other had poor phonological awareness, produced relatively few phonologically accurate spelling errors, and was poor at reading and spelling nonwords (i.e. was a phonological dyslexic). It is particularly noteworthy that such a clear dissociation between surface and phonological forms of developmental dyslexia occurred in two subjects who were closely matched in terms of their overall reading and spelling ability, and also in terms of their memory span and vocabulary. It is argued that this study strengthens the evidence for the existence of qualitatively different types of developmental dyslexia. The findings are also consistent with the view that phonological awareness skills are more closely related to the operation of the phonological rather than the visual reading route. PMID- 7477819 TI - Interneuronal functional associations in the sensorimotor cortex of dogs. AB - The interneuronal functional associations were studied in two dogs with Nichrome semi-microelectrodes implanted into the deep layers of the motor and somatosensory regions of the cerebral cortex using the method of cross correlation analysis. For this purpose the impulse activity of individual neurons was distinguished by form from the background multineuronal activity using the spike recognition technique. Values of a 0.5 and 1 msec-wide bin, and thereafter with a 1 msec step up to 40 msec, were used to plot the cross-interval histograms. The maximal analysis epoch was 2000 msec. The cross-interval associations were monotypal in character; they all presented fairly narrow extrema which were clearly distributed across three time ranges: short-latency associations up to 10 msec; associations with a medium latency up to 80 msec; and associations with late delays, greater than 80 msec. The fairly narrow peak of the association, especially in the case of associations with late delays, was a very difficult phenomenon to explain from the point of view of traditional theoretical perspectives. It is hypothesized that a mechanism exists in the cortex which is responsible for strictly synchronized and highly efficient synaptic transmission. PMID- 7477818 TI - Participation of the anterior suprasylvian cortex of the cat in the preparation of the paw extension reaction. AB - The electrical stimulation of the anterior suprasylvian cortex of cats (field 5b) induces the contralateral forepaw extension reaction. Neurons were recorded in the focus of effective stimulation which were tonically activated under the influence of motivationally significant visual and tactile stimuli which anticipate or elicit the reactions of paw extension and/or paw placement on a support. PMID- 7477821 TI - Activity of neurons of the cat motor cortex during differentiation between reactions of right and left paw placement on a support developed in response to stimulation of the parietal cortex of the different hemispheres. AB - Differentiation was developed in cats between reactions of the placement of the right and left paw on a support in response to stimulation of the ipsilateral (right or left, respectively) parietal cortex (PC) in the region of field 5. The shortest-latency primary responses of the neurons of the MC do not vary in relation to whether stimulation of the PC does or does not elicit the conditioned motor reaction of the limb. Stimulation of the PC as a conditional signal triggering the cPR elicits, in addition to the primary responses, the generation of excitatory reactions of neurons of the MC in the interval from 20 to 200 msec and longer; if the motor reaction is not triggered or if a reaction of the limb ipsilateral to the motor cortex is triggered, an inhibitory pause is generated in the indicated time interval. PMID- 7477820 TI - Long-term sensitization in the common snail: electrophysiological correlates in the defense behavior command neurons. AB - It was found that an increase in the excitability of command neurons of the reflex takes place during long-term sensitization (LTS). It was demonstrated that two factors are the cause of the increase in excitability: a depolarizational shift of the membrane potential of these cells (both a decrease in the initial resting potential and an accumulation of depolarization in the course of tactile stimulation), and a decrease in the threshold of generation of the action potentials. Injection of amphetamine leads to the appearance of sensitization, but its mechanisms only partially coincide with those in the case of LTS. PMID- 7477822 TI - Role of kynurenines in the regulation of the honeybee's memory process. PMID- 7477823 TI - Investigation of the stages of the mental rotation of complex figures with the intracortical interaction mapping technique. AB - The stages of the mental rotation of complex three-dimensional figures were investigated in nine healthy subjects using the intracortical interaction mapping technique. It was established that mental rotation is accomplished with the participation of both parietal regions; the prerotation setup is associated with the involvement of the frontal, central, and right temporal divisions of the brain; decision-making regarding the result of the rotation and the response is accomplished through the conjunction of both frontal and temporoparietooccipital regions of the left hemisphere. In the case of the unsuccessful solution of the problem the process seems to stop at the stage of the prerotation setup. With verbal control the configuration of the zone of high-level association during the first three stages is reminiscent of that observed in the baseline activity. PMID- 7477824 TI - Characteristics of the chemical sensitivity of cerebral cortical neurons at the stages of food-procuring behavior of cats as a function of the quality of reinforcement. AB - The chemical sensitivity to the principal neuromediators of neurons of the visual area of the cerebral cortex was investigated at sequential stages of instrumental behavior in this study using the method of the microiontophoresis, in two experimental situations. In the first of these a portion of milk was used as the reinforcement; in the second, a portion of meat powder. It was demonstrated that the number of universal neurons decreases against the background of the application of norepinephrine and acetylcholine in behavioral acts with qualitatively different food reinforcements. At the same time, the number of neurons that are areactive at all stages of the behavior increases in the situation involving milk reinforcement, while it decreases in the situation involving meat reinforcement. PMID- 7477825 TI - Motivational support of complex behavioral reactions in rats. AB - The activity of neurons of the medial wall of the frontal cortex (MWFC) of the brain of rats trained to a delayed spatial choice task was investigated. Significant reorganizations of the activity of the neurons were found when the motivational state of the animal was changed. Direct connections were identified between hypothalamic nuclei and the MWFC using the retrograde axonal horseradish peroxidase transport method. The role of hypothalamocortical interactions in providing the motivational-emotional components of behavior is discussed. PMID- 7477826 TI - Neurotransmitter basis of mollusc behavior: control of choice between the orienting and the defense response to the presentation of an unfamiliar object. AB - The role of the serotoninergic and opioid systems in the choice between a passive defensive and an orienting response of the mollusc Lymnaea stagnalis to the presentation of an unfamiliar object was studied. Under the influence of 5-HTP, a metabolic precursor of serotonin, orienting and investigatory behavior was activated; the number of protective reactions decreased. Under the influence of naloxone, an antagonist of opiate receptors, the proportion of orienting responses decreased, and the number of passive defensive avoidance and freezing reactions increased. The influence of the agents investigated on the attitude of the snail to a novel object was found to be well coordinated with their influence on other forms of behavior and the behavioral state of the mollusc as a whole. PMID- 7477827 TI - Physiological problems of biofeedback control of the pulse rate. AB - Data are presented in this article on the voluntary control of the heart rate in man as a function of individual typological differences in adaptive attributes, in the ecological features of the milieu of habitation and work, as well as in the presence of deviations in the state of health (neuroses, vegetovascular dystonias, etc.). The key significance of training of the rhythmic regulatory processes in the course of biofeedback control is substantiated. PMID- 7477828 TI - Emotional stress and extracardiac regulation. AB - It has been demonstrated in chronic experiments on animals (rabbits, rats) that in the presence of a predominance of sympathetic influences on the heart during emotional stress, the disturbances that appear in metabolism and myocardial ultrastructure promote a decrease in the electrical stability of the heart, the development of disturbances in cardiac rhythm, and arterial hypertension. A predominance of parasympathetic influences on the heart during stress, to the contrary, leads to an increase in the electrical stability of the heart, and also impedes an increase in the content of catecholamines and the development of structural injuries in the myocardium. At the same time, arterial hypertension is absent. An inference is drawn regarding the adaptive-trophic, protective action of the vagus nerves on the heart during emotional stress. PMID- 7477829 TI - Combinations of methods for monitoring the microcirculation of the brain. PMID- 7477830 TI - Postmortem MRI of the brain with neuropathological correlation. AB - We prospectively correlated in vitro MRI of the brain with the neuropathological findings in patients with suspected intracranial disease. In vitro MRI was performed on 91 consecutive formalin-fixed whole-brain specimens. In 60 cases, the images were correlated with the neuropathological findings (number of lesions, lesion boundaries, spread of oedema and type of lesions). As compared with neuropathology, MRI showed an equal number of lesions in 50 cases, more in 5, and less in 5 specimens, resulting in a sensitivity of 83.3%. The extent of perilesional oedema was better seen on in vitro MRI than on gross pathology. Microscopic extent of glial tumours was underestimated on both T2-weighted images and macroscopic examination. Neuropathology remains the reference study, since on in vitro MRI primary brain tumours, metastatic deposits and non-neoplastic space occupying lesions cannot be differentiated. However, in our study MRI had a specificity of 76.6%. MRI of postmortem specimens is sensitive to focal brain lesions, and can focus the attention of the neuropathologist to abnormal regions. PMID- 7477831 TI - Three-dimensional-MRI of neurovascular compression in patients with hemifacial spasm. AB - We prospectively studied 30 patients to assess the usefulness of three dimensional (3D) contrast-enhanced MRI in patients with hemifacial spasm. In all patients neurovascular compression of the facial nerve could be detected. Microvascular decompression was performed in 14, and vascular compression of the root exit zone of the facial nerve and the offending artery were exactly as predicted by MRI in all but 2. We also retrospectively studied whether the symptomatic side could be defined only by enhanced 3D MRI in 55 randomised individuals (30 with and 25 without hemifacial spasm). All symptomatic sides were correctly identified, and the false-positive rate was 13.8%. PMID- 7477832 TI - Magnetic resonance tomographic angiography: diagnostic value in trigeminal neuralgia. AB - A combination of MRI, MR angiography and MR tomographic angiography (MRTA) was used to study the relationship of the root exit zone of the trigeminal nerve to surrounding vascular structures in seven patients with trigeminal neuralgia (TN) and ten patients with no evidence at a lesion in this region. MRTA is the technique for showing the relationship between vessels, cranial nerves and brain stem. MRTA clearly demonstrated the presence of a vessel at the root exit zone of the trigeminal nerve in all patients with TN. In the ten other patients, examination of 20 trigeminal nerves revealed that only one nerve (5%) was in contact with a vessel at the root exit zone. This study supports vascular compression of trigeminal nerves as a cause of TN, and demonstrates the value of MRTA as noninvasive technique for demonstrating compression. PMID- 7477833 TI - Moyamoya disease: diagnostic accuracy of MRI. AB - Our purpose was to evaluate the diagnostic accuracy of MRI in moyamoya disease. We studied 30 patients with this disease, comparing MRI and angiographic findings. The diagnostic value of MRI was evaluated for occlusive lesions, collateral vessels, and parenchymal lesions. In all patients bilateral occlusion or stenosis of the supraclinoid internal carotid artery and proximal anterior and middle cerebral arteries was clearly shown by MRI, and staging of the extent of occlusion agreed with angiographic staging in 44 (73%) of 60 arteries. MRI, particularly coronal images, clearly showed basal cerebral moyamoya vessels in 54 hemispheres, and 45 of a total of 71 large leptomeningeal and transdural collateral vessels were identified. MRI also showed parenchymal lesions in 48 (80%) hemispheres, and the extent of occlusion in the anterior and posterior circulations respectively correlated with white matter and cortical and/or subcortical infarcts. PMID- 7477834 TI - Hypoplasia of the internal carotid artery associated with spasmodic torticollis: the possible role of altered vertebrobasilar haemodynamics. AB - We describe an unusual case of hypoplasia of the left internal carotid artery associated with an anomalous origin of the ipsilateral middle cerebral artery in a patient with spasmodic torticollis. Collateral circulation to the territory of the middle cerebral artery was through the vertebrobasilar circulation, and the left vertebral artery was dilated and compressing the ventrolateral aspect of the cervicomedullary junction, as evidenced by MRI. We believe this to be the first report of unilateral hypoplasia of the internal carotid artery associated with spasmodic torticollis. Compression of the left accesory nerve by the dilated vertebral artery as a result of altered haemodynamics was thought to cause spasmodic torticollis. PMID- 7477835 TI - 99mTc HM-PAO brain perfusion SPECT in brain death. AB - Diagnosis of brain death must be certain to allow discontinuation of artificial ventilation and organ transplantation. Brain death is present when all functions of the brain stem have irreversibly ceased. Clinical and electrophysiological criteria may be misinterpreted due to drug intoxication, hypothermia or technical artefacts. Thus, if clinical assessment is suboptimal, reliable early confirmatory tests may be required for demonstrating absence of intracranial blood flow. We have easily carried out and interpreted 99mTc HM-PAO SPECT in a consecutive series of 40 comatose patients with brain damage, without discontinuing therapy. Brain death was diagnosed in 7 patients, by recognising absence of brain perfusion, as shown by no intracranial radionuclide uptake. In patients in whom perfusion was seen on brain scans, HM-PAO SPECT improved assessment of the extent of injury, which in general was larger than suggested by CT. PMID- 7477836 TI - Occipital condyle fractures diagnosed by high-definition CT and coronal reconstructions. AB - We describe four cases of fracture of the occipital condyle. In all cases plain cervical spine films were normal. The fracture was diagnosed on the lowest section of a routine CT examination of the head. Further detail was then obtained with high-definition CT and coronal reconstructions. All the patients were found to have a small atlanto-occipital subluxation, no greater than 11 mm. Classification and treatment of these fractures is discussed. It is stressed that the lower-most section of a CT head study for cerebral trauma should include the inferior margin of the foramen magnum and that both bone and soft tissue images should be obtained. PMID- 7477837 TI - Serial MRI changes in radiation myelopathy. AB - Using MRI we assessed the changes in signal, size, and contrast enhancement characteristics of the cervical spinal cord in radiation myelopathy developing after radiotherapy for nasopharyngeal carcinoma. We studied two men and five women, aged 40-77 years. The first MRI study was performed 1-4 months after the initial clinical manifestations of myelopathy, and follow-up MRI 2-22 months after the onset of symptoms. On the first study, all patients showed low signal intensity in a long segment of the cervical spinal cord on T1-weighted images, high signal on T2*-weighted images, and focal contrast enhancement at C1-2. In five patients there was also swelling of the spinal cord. The site of eccentric focal contrast enhancement correlated with the clinical manifestations. Follow-up imaging less than 10 months after the onset of symptoms showed no significant changes in signal intensity. Focal contrast enhancement at C1-2 remained the same in three patients, was more dense and larger in one, and less dense in another. Subsidence of swelling was seen in two patients. Atrophy of the spinal cord at C1 2, without abnormal signal and with faint contrast enhancement at C1-2 was revealed as early as 10 months after the onset of symptoms, but the contrast enhancement disappeared by 22 months. There was no correlation between clinical manifestations and spinal cord atrophy on MRI. PMID- 7477838 TI - Serial changes on MRI in lumbar disc herniations treated conservatively. AB - Using serial MRI, we studied 32 patients with herniated lumbar discs, treated conservatively, to clarify the natural history of this condition. MRI was performed in the acute stage, then 6 months and 1 year later. On axial images, the proportion of the cross-sectional area of the spinal canal occupied by the herniated disc was 31.9% on the average on the initial scan, 28.7% 6 months and 25.3% 1 year later. The size of the herniation decreased by more than 20% in 11 patients (34%), by 10-20% in 8 (28%) and was unchanged in 12 (38%). The height of the disc slightly decreased with time, but there was no significant change in the angle of lordosis in the affected segment. The initial MRI revealed degeneration of all affected discs, and progressive degeneration was observed in 9 patients. The more degenerate the disc and the larger the initial herniation the more the size of the herniated fragment decreased. PMID- 7477839 TI - MRI in the investigation of patients with myelopathy thought to be due to multiple sclerosis. AB - The role of cerebral and spinal cord MRI was investigated in 65 patients with myelopathy suspected of having demyelinating disease. Cerebral MRI demonstrated lesions compatible with demyelination in 80% and spinal cord MRI in 68.6%. In 28.5% of our patients brain lesions were present with normal spinal cord images, but in 17% spinal cord lesions were depicted with a normal brain MRI. The combination of the two examinations demonstrated lesions in 97% of the patients. The frequency of coexistent cerebral lesions in patients with spinal cord lesions was over 85% in patients with chronic disease but only 28.5% in patients with acute myelitis. PMID- 7477840 TI - CT and MRI of the normal and diseased perivertebral space. AB - The perivertebral space is in the midline, in the deep tissues of the neck, and can be identified from the skull base above to the mediastinum below. It is a discrete space completely enclosed by the deep layer of deep cervical fascia. The fascial attachments of the perivertebral space divide it into two areas, the anterior prevertebral and posterior paraspinal portions. We made a retrospective analysis of the radiologic and clinical records of 52 patients with lesions in the perivertebral space, to identify the imaging features that mark a lesion as originating in the perivertebral space and define the spectrum of pathology which occurs in the space. Mass lesions present in the prevertebral or paraspinal portions. In the former they usually involve the vertebral body, displacing the prevertebral muscles anteriorly. Epidural extension from lesions in the perivertebral space proper is common. Masses in the paraspinal perivertebral space usually displace the paraspinal muscles away from the spine. We found 9 inflammatory lesions, 29 malignant and 6 benign tumors, and 8 miscellaneous lesions. PMID- 7477841 TI - Intravascular ultrasonography of supra-aortic arteries as an adjunct to percutaneous transluminal angioplasty. AB - We report the intravascular ultrasonographic appearances of three supra-aortic atherosclerotic lesions in two patients before and after percutaneous transluminal angioplasty. Atherosclerotic plaques with calcification and dissection of the arterial wall after percutaneous transluminal angioplasty were demonstrated to better advantage, although they were difficult to see on conventional angiograms. PMID- 7477842 TI - Percutaneous transvenous embolisation of iatrogenic vertebral arteriovenous fistula. AB - Two patients presented with vertebral arteriovenous fistulae following unintentional puncture of the vertebral artery. A percutaneous transvenous approach was used in both cases and the fistula was successfully embolised with microcoils. A complete cure was achieved in both patients. PMID- 7477843 TI - Hyperhomocysteinaemia; with reference to its neuroradiological aspects. AB - Severe or even mild hyperhomocysteinaemia can cause a wide range of neurological problems. In recent years its vascular complications, including cerebral stroke, in children and young adults have gained special interest, because hyperhomocysteinaemia is treatable and recurrence of vascular incidents may be preventable. Current knowledge about biochemical mechanisms leading to hyperhomocysteinaemia, the pathogenesis of vascular pathology and neurological disfunction, and the various patterns of cerebral damage are reviewed. The significance of MRI in diagnosis, follow-up and research on hyperhomocysteinaemia is discussed. PMID- 7477844 TI - A characteristic ventricular shape in myelomeningocele-associated hydrocephalus? A CT stereology study. AB - We measured the volume of the supratentorial ventricles in 39 consecutive children with myelomeningocele (MMC) and associated hydrocephalus, using a stereological method based on the Cavalieri theorem of systematic sampling. We distinguished the following groups: newborns before and after cerebrospinal fluid shunting (14), a somewhat larger group of newborns with an untreated MMC associated hydrocephalus (25) and a group of shunted children at a mean age of 1.5 years (28). We paid special attention to the shape of the lateral ventricles, looking separately at the anterior and posterior halves. The measurements were compared with a healthy control group (10) and with children with hydrocephalus unrelated to MMC (15). The average volume ratio of the posterior to the anterior half of the lateral ventricles was 1.05 +/- 0.39 in non-hydrocephalic children, 1.11 +/- 0.55 in untreated hydrocephalic children without MMC, and 2.15 +/- 0.65 in MMC-associated hydrocephalus prior to shunting. These ratios did not change significantly after shunting. This confirms our impression that MMC-associated hydrocephalus shows a characteristic shape, with a disproportionate enlargement of the posterior part of the lateral ventricles, in clear contrast to the normal width frontal horns. This shape is reminiscent of the fetal ventricular shape. It reveals disturbance of brain development in children with MMC, which goes beyond the classic description of the Chiari malformation. PMID- 7477846 TI - Serial MRI of olivary hypertrophy: long-term follow-up of a patient with the "top of the basilar" syndrome. PMID- 7477845 TI - The size of the intra- and extraventricular cerebrospinal fluid compartments in children with idiopathic benign widening of the frontal subarachnoid space. AB - The aim of this study was to quantify the intra- and extraventricular cerebrospinal fluid (CSF) spaces in children with benign enlargement of the frontal subarachnoid space (BE). The infra- and supratentorial CSF compartments were measured in 61 CT examinations of children with BE, 3-27 months old, and compared with those of 96 CT examinations considered normal. Measurements of the ventricular system, and the pontine and chiasmatic cisterns were related to cranial size. In all children with BE the lateral and third ventricles were dilated and the chiasmatic cistern was widened. The subarachnoid space was wider than the upper limits in the control group, in the frontal region (4 mm), and the anterior interhemispheric (4 mm) and Sylvian (3 mm) fissures. The infratentorial CSF compartments, the occipital subarachnoid space, the posterior part of the interhemispheric fissure and, in most cases, the cortical sulci were normal in size in children with BE. The majority were macrocephalic or had rapid head growth but there were also normocephalic children with normal head growth. The size of the posterior fossa was within the normal range in all children with BE. Idiopathic BE is not uncommon in children up to about 3 years old who are healthy or have minimal neurological disturbance and is characterised by a specific pattern of widening of the supratentorial CSF compartments. PMID- 7477847 TI - Spatial data analysis in the quantitative assessment of cerebral white matter pathology on MRI in HIV infection. AB - This study was carried out using MRI (proton density--and T2-weighted) on 16 HIV negative controls, 9 symptom-free HIV-positive patients and 25 with CDC IV HIV disease. The studies from this last group had previously been allocated by a radiologist to the following categories: 8 with focal mass lesions and normal appearing white matter; 9 with diffuse encephalopathy (high signal on T2-weighted images, affecting most or all of the white matter) and 8 with patchy encephalopathy (high signal affecting only one or two areas within the white matter). Moran's I, a statistic of spatial autocorrelation, was calculated for the grey-scale values of a sampled pixel array from a central white matter region of each of the images. All values of Moran's I calculated in this study showed a large positive excess over the expected value under randomisation, indicating highly significant positive auto-correlation in the spatial arrangement of the grey-scale values. On T2-weighted images a statistically significant increase in the mean value of Moran's I, compared with controls, was found in the diffuse encephalopathy group, indicating that quantifiable changes in the spatial autocorrelation of pixel data can be related to recognised qualitative changes in the appearance of white matter in subjects with HIV disease. A lesser, but significant, rise in the mean value of Moran's I was also found in the focal mass lesion group, suggesting that changes in spatial autocorrelation may indicate pathological change in advance of qualitative MRI changes. PMID- 7477848 TI - Acute disseminated encephalomyelitis (ADEM) or ADEM-like inflammatory changes in a heart-lung transplant recipient: a case report. AB - Acute disseminated encephalomyelitis (ADEM) is a white matter inflammatory disease which usually follows a viral infection or pharmaceutical intervention. We describe a case of presumed ADEM in a heart/lung transplant patient, the etiology of which cannot be elucidated. The fascinating aspect is the mode of clinical presentation and the rapid resolution of radiologic abnormalities. Histologic examination of the brain is provided in an attempt to elucidate the radiographic abnormalities. PMID- 7477849 TI - Status epilepticus presenting as progressive dysphasia. AB - Status epilepticus is usually a straightforward diagnosis when a patient has two or more seizures without regaining consciousness. However, when status is non convulsive and, in particular, has a temporal lobe flavour the clinical presentation may be misleading. Presentation with automatic or psychic behaviour is well recorded. We report a patient with nonconvulsive status who presented with progressive dysphasia with widespread CT and MRI changes. The dysphasia and imaging changes led to a diagnosis of a probable neoplastic brain process but reversed with anticonvulsant treatment. PMID- 7477850 TI - Functioning heterotopic grey matter? Increased blood flow with voluntary movement and sensory stimulation. AB - A 66-year-old woman with a right hemiplegia due to cerebral haemorrhage was found to have heterotopic grey matter in the left frontal and right frontal and parietal lobes. She had no neurological disturbance prior to the haemorrhage. Regional cerebral blood flow (rCBF) was measured by xenon-CT (Xe-CT) in the zones of heterotopic grey matter during voluntary movement and sensory stimulation of her left limbs. During voluntary movement, rCBF increased markedly in the heterotopic grey matter of the frontal lobes and the overlying cortex. During sensory stimulation, rCBF increased in the right parietal heterotopic grey matter and the overlying cortex. This strongly suggests that the heterotopic grey matter is functional. PMID- 7477851 TI - Cranial MRI in chronic thinner intoxication. AB - We studied a 19-year-old man with thinner and toluene poisoning for 5 years by CT and MRI. Symmetrical lesions were seen in the basal ganglia and cingulate gyri. PMID- 7477852 TI - MRI in chronic paint-thinner intoxication. PMID- 7477853 TI - MRI in internal cerebral vein thrombosis: case note. AB - A 40-year-old man developed general malaise, dizziness and progressive headache 1 week after acute haemorrhagic conjunctivitis. Bizarre behaviour, confusion and adversive seizures occurred later. MRI showed haemorrhagic infarct in the bilateral basal ganglia and thalamus and abnormal signal density in the internal cerebral veins. Bilateral carotid angiography showed no filling of superior sagittal sinus and internal cerebral veins. With MRI, cerebral venous thrombosis can be suspected and diagnosed earlier than before, obviating any invasive investigation. PMID- 7477854 TI - MRI of a pituitary cryptococcoma simulating an adenoma. AB - We describe a rare pituitary cryptococcoma in an immunocompetent patient, with radiological features similar to those of a pituitary macroadenoma. Although unusual, it should be added to the list of differential diagnosis of pituitary masses. Contrast enhancement of adjacent meninges differentiated the lesion from an adenoma. PMID- 7477855 TI - Granular cell tumour of the neurohypophysis. AB - Granular cell tumours of the neurohypophysis are very rare. Their aetiology and nature are uncertain. Most investigators consider them to arise from pituicytes or Schwann cells of the posterior lobe of the hypophysis. They are found most commonly in older people. A surgically proven symptomatic case is reported. PMID- 7477856 TI - MRI in spontaneous cerebrospinal fluid rhinorrhoea. AB - We studied two patients with spontaneous cerebrospinal fluid (CSF) rhinorrhoea with MRI and other imaging modalities. T2-weighted images proved most useful for the detection and localisation of the CSF leakage. MRI appeared to provide an accurate and noninvasive method for preoperative investigation of spontaneous CSF rhinorrhoea. PMID- 7477857 TI - Transcalvarial primary lymphoma of bone. A report of two cases. AB - Primary lymphoma of bone is uncommon, and initial involvement of the calvarium is rare. We describe two cases originating as enlarging skull masses subsequently shown to involve the bony calvarium, extracranial soft tissues, and intracranial structures. Open biopsy revealed non-Hodgkin's lymphoma, large cell type, in both cases. A review of the pertinent literature follows. PMID- 7477858 TI - Idiopathic intracranial hypertension: orbital MRI. AB - The case is reported of a patient with idiopathic intracranial hypertension examined with magnetic resonance imaging. Marked enhancement of the optic nerve heads was found, which might be related to blood-retinal barrier breakdown related to a sudden rise in intracranial cerebrospinal fluid pressure. PMID- 7477859 TI - Septic arthritis in a lumbar facet joint: a rare cause of an epidural abscess. AB - A 10-year-old boy presented with a 7-day history of back pain and pyrexia. MRI showed an epidural abscess arising from septic arthritis in a lumbar facet joint. Whilst septic arthritis in a large joint is relatively common in children, epidural abscess is rare. This case illustrates how infection in a synovial joint may extend into the extradural space and might be the route of infection in more cases than has previously been recognised. PMID- 7477860 TI - Primary leiomyosarcoma of the spine. AB - We present a 39-year-old man with tumour of the eighth thoracic vertebra, causing compression of the spinal cord. The tumour proved to be a primary leiomyosarcoma (LMS) of bone, an uncommon neoplasm; to our knowledge this is the first report of primary LMS in the spine. The lesion was documented by plain radiography, myelography, CT, MRI and digital subtraction angiography. These investigations did help to focus on the differential diagnosis and demonstrated the extent of the bony lesion, the findings were nonspecific, and the correct diagnosis was established by pathological examination. PMID- 7477861 TI - Calcification of cervical ganglia: case report. AB - We describe a 44-year-old woman with progressive cervical pain in whom plain films of the cervical spine showed only minor syndesmophyte formation when the patient first presented. However, after 5 weeks, repeat films demonstrated heavy symmetrical calcification of the cervical dorsal root ganglia. A review of the literature did not reveal a previous description of these findings. PMID- 7477862 TI - Changes in paranasal sinus abnormalities found incidentally on MRI. AB - Abnormal opacification of paranasal sinuses on brain MRI was studied in 178 consecutive adult patients who underwent two examinations separated by at least 4 months because of suspected neurological disease. Sinus abnormality was present in 79 (44%) patients; in 60% the abnormality involved two or more sinuses and it was bilateral in 51%. The most frequently abnormal sinus was the maxillary (46%), followed by the ethmoid (32%), sphenoid (16%), and frontal sinuses (6%). Interval occurrence or complete resolution of abnormalities was observed in 52% of patients (66% of the sinuses). In the remaining 48% of patients the abnormalities were present on two or more examinations, unchanged or of increased or decreased severity. The most frequent interval changing abnormalities were complete sinus opacification and fluid levels, followed by irregular mucoperiosteal thickening more than 4 mm thick. No polypoid abnormalities (polyps or mucosal cysts) decreased over time. PMID- 7477863 TI - MRI of idiopathic orbital inflammatory syndrome using fat saturation and Gd-DTPA. AB - Idiopathic orbital inflammatory syndrome encompasses a group of inflammatory conditions for which no systemic or local cause can be found, and is commonly referred to as orbital pseudotumour. On conventional MRI sequences subtle areas of inflammation or enhancing tissue can easily be masked by the high signal intensity of orbital fat and involvement of the fat itself may not be appreciated. We describe the MRI features of three patients with idiopathic orbital inflammation using frequency-selective fat saturation and Gd-DTPA. PMID- 7477864 TI - Meningioma presenting as a mass in the internal auditory canal. PMID- 7477865 TI - Gliofibroma: CT and MRI. AB - We describe CT and MRI appearances in two children with pathologically proven gliofibromas, in the cerebrum and cerebellum. A striking finding was lack of high signal on T2-weighted MRI. PMID- 7477866 TI - Dynamic enhancement MRI of anterior lobe in pituitary dwarfism. AB - We examined 23 patients with pituitary dwarfism by dynamic MRI; with a repetition time of 150 or 50 ms. The time-enhancement difference curves of selected regions in the anterior lobes were plotted. Another 48 patients with no definite clinical pituitary disfunction were examined with the same technique. We found that the intensity of maximum enhancement in both groups was similar, but the time to achieve maximum enhancement was delayed in pituitary dwarfism with or without stalk transection; the time seemed longest with stalk transection. There was little difference in enhancement between patients with multiple hormone deficiency or isolated growth hormone deficiency. Dynamic MRI of the anterior lobes may be an important functional imaging study, and our results imply that poor perfusion is a useful finding in pituitary dwarfism, especially in patients without stalk transection and normal pituitary height. PMID- 7477867 TI - Neuroradiological findings in the carbohydrate-deficient glycoprotein syndrome. AB - The carbohydrate-deficient glycoprotein syndrome is a newly recognised genetic disorder characterised by mental retardation, liver disfunction during infancy, cerebellar ataxia and atrophy, polyneuropathy, growth retardation, stroke-like episodes, and the appearance of carbohydrate-deficient fractions of multiple glycoproteins in the serum. The neuroradiological findings have been known as features of olivopontocerebellar atrophy. However, whether the abnormalities in the cerebellum and brain stem progress after birth is not known. We have carried out serial CT and MRI on three Japanese patients with this syndrome at different ages. A small cerebellum, with peculiar enlargement of the cisterna magna, and a small brain stem are present in infancy and atrophy of the anterior vermis and from before backwards in the cerebellar hemispheres seem to progress throughout early childhood. PMID- 7477868 TI - MRI of peripheral nerves and pathology of sural nerves in hereditary motor and sensory neuropathy type III. AB - We investigated two patients with hereditary motor and sensory neuropathy type III, one with Dejerine-Sottas disease and the other with congenital hypomyelination neuropathy based on nerve pathology and MRI of the sciatic nerve. On biopsy of the sural nerve of the patient with Dejerine-Sottas disease, myelin debris, indicating demyelination, was observed in an onion-bulb pattern surrounding myelinated fibres. In the patient with congenital hypomyelination neuropathy, onion bulbs were formed of two parallel layers of basement membrane. There was no evidence of myelin breakdown. On axial T2-weighted MRI, a severely hypertropied sciatic nerve containing multiple rounded lesions, suggesting inflammation or demyelination, was observed in the patient with Dejerine-Sottas disease. In contrast, the sciatic nerve of the patient with congenital hypomyelination neuropathy showed slight hypertrophy without demyelination. MRI of the sciatic nerve may represent a useful tool for characterisation of demyelinating disease and its prognosis. PMID- 7477869 TI - Stress-induced changes in messenger RNA levels of N-methyl-D-aspartate and AMPA receptor subunits in selected regions of the rat hippocampus and hypothalamus. AB - The postsynaptic AMPA/kainate and N-methyl-D-aspartate-selective glutamate receptors are formed by several different subunits and the overall subunit composition of the receptor appears to determine its physiological and pharmacological properties. Although glutamatergic mechanisms have been implicated in various forms of hippocampal stress responses, the impact of stress on glutamate receptor subunit composition has not yet been elucidated. We have used cell-by-cell quantitative in situ hybridization to assess stress-induced changes in transcript levels of N-methyl-D-aspartate and AMPA receptor subunit genes in subdivisions of the rat hippocampus and hypothalamus that are implicated in the stress response. We found that 24 h after a single immobilization stress there was a significant increase in the cellular level of NR1 subunit messenger RNA (about 35-45% above control values) in hippocampal CA3 and CA1 pyramidal cells as well as in neurons of the hypothalamic supraoptic and paraventricular nuclei. Moreover, in the CA3 area we have detected a concomitant increase (50% above controls) in the level of NR2B subunit messenger RNA, while the expression of NR2A subunit gene did not change after stress. Stress induced a selective decrease in the level of AMPA receptor subunit glutamate receptor A messenger RNA in neurons of both the CA3 and CA1 areas (18 and 24%, respectively, below control values). These results suggest that the regulation of specific subunit messenger RNAs of the N-methyl-D-aspartate and AMPA receptors may be involved in altered hippocampal and hypothalamic responsiveness to glutamate and thus could play a critical role in stress-induced changes in their function. PMID- 7477870 TI - Corticothalamic projections from the primary visual cortex in rats: a single fiber study using biocytin as an anterograde tracer. AB - This study investigates the pattern of axonal projections of single corticothalamic neurons from the rat primary visual cortex. Microiontophoretic injections of biocytin were made in cortical laminae V and VI to label small pools of corticothalamic cells and their intrathalamic axonal projections. After a survival period of 48 h, the animals were perfused and the tissue was processed for biocytin histochemistry. On the basis of the intrathalamic distribution of axonal fields and the types of terminations found in the thalamus, three types of corticothalamic projections were identified. (1) Cells of the upper part of lamina VI projected to the dorsal lateral geniculate nucleus where they arborized in rostrocaudally oriented bands or "rods" parallel to the lines of projection of retinal afferents. (2) Cells of the lower part of lamina VI projected to the lateral part of the lateral posterior nucleus and they also sent collaterals to the dorsal lateral geniculate nucleus where they participated in the formation of rods. (3) The corticothalamic projection of lamina V cells originated from collaterals of corticofugal cells whose main axons reached the tectum and/or the pontine nuclei. These collaterals never terminated within the dorsal lateral geniculate nucleus; they arborized in the lateral posterior, lateral dorsal and ventral lateral geniculate nuclei. All corticothalamic cells from lamina VI displayed the same type of axonal network made of long branches decorated by terminal boutons emitted "en passant" at the tip of fine stalks. Corticothalamic fibers arising from lamina V, however, generated varicose endings in restricted regions of their target nuclei. All corticothalamic axons derived from lamina VI cells, but not those derived from lamina V cells, gave off collaterals as they traversed the thalamic reticular complex. These results demonstrate that corticothalamic fibers arising from the rat primary visual cortex display a lamina-dependent projection pattern. In the light of previous studies on the topographical organization of corticothalamic projections, it is proposed that a similar organizational plan characterizes corticothalamic relationships in other sensory systems in the rat and in other species. PMID- 7477871 TI - Susceptibility of different cell layers of the anterior and posterior part of the piriform cortex to electrical stimulation and kindling: comparison with the basolateral amygdala and "area tempestas". AB - Several lines of evidence suggest that the piriform cortex functions as a generator in the development and propagation of forebrain (limbic type) seizures, particularly in the kindling model of epilepsy. It is, however, not clear where, within the rather large piriform cortex region, the generator resides, and how much tissue is involved. Highly sensitive loci to chemical or electrical stimulation have been described both in the deep anterior and posterior parts of the piriform cortex. Furthermore, data from piriform cortex slice preparations indicated that epileptiform potentials originate in deep structures, particularly the endopiriform nucleus that underlies the piriform cortex. In the present study, in rats, we implanted stimulation and recording electrodes in various rostrocaudal locations of the piriform cortex and endopiriform nucleus, including the "area tempestas", i.e. a structure in the anterior part of the piriform cortex previously proposed to be critically involved in the generation of convulsive seizures of limbic origin. Within the piriform cortex, electrodes were aimed at different cellular layers of this structure. For comparison, additional animals received electrodes in different parts of the basolateral amygdala. A total of 19 different locations was obtained in this way. The susceptibility of these locations to electrical stimulation was characterized by determining the threshold for induction of afterdischarges. The afterdischarge threshold was lowest in layer III of the posterior piriform cortex and some locations in the endopiriform nucleus, whereas amygdala and "area tempestas" displayed higher values. In several animals, particularly those with electrodes in layer III of the posterior piriform cortex, spontaneous spiking was seen in prestimulation recordings, whereas this was never observed in recordings from the amygdala. Subsequent kindling by repeated stimulation of the various locations demonstrated marked differences in afterdischarge threshold reduction and kindling rate. The most marked decreases in afterdischarge threshold were seen in locations within layer III of the piriform cortex, whereas several other locations, including the "area tempestas", exhibited only moderate decreases or no decrease at all. In contrast to previous observations with only few locations in the piriform cortex region, the posterior piriform cortex was not in general slower to kindle than the anterior piriform cortex, although some locations in the posterior piriform cortex exhibited significantly lower kindling rates than the amygdala. The highest kindling rate was seen in the dorsal endopiriform nucleus.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 7477872 TI - Comparison of long-term potentiation in the proximal versus distal stratum radiatum of hippocampal field CA1. AB - Recent studies indicate that long-term potentiation is accompanied by changes in the waveform of field excitatory postsynaptic potentials in the CA1 field of disinhibited hippocampal slices, suggesting that long-term potentiation alters the kinetics of the glutamate receptor channels that mediate excitatory synaptic responses. The present study examined the effects of stimulating and recording location within stratum radiatum on the magnitude of long-term potentiation and the associated waveform changes. Stimulation of stratum radiatum proximal to the cell body layer evoked field excitatory postsynaptic potentials in proximal stratum radiatum that had decay phases well-fit by single exponentials; long-term potentiation reduced the decay time constant of these potentials, as reported previously. Stimulation of distal stratum radiatum evoked field excitatory postsynaptic potentials in distal stratum radiatum that were contaminated by positive after-potentials; meaningful estimates of decay time constants for these responses could not be obtained. Long-term potentiation of distal responses tended to be smaller than that obtained proximally. Comparisons were also made between responses recorded distally and proximally for either distal or proximal stimulation. For both stimulation loci, the distally-recorded responses had positive after-potentials and the proximally-recorded responses did not. The decay time constants for proximally-recorded responses to distal stimulation decreased significantly after long-term potentiation. Proximal and distal stimulation both produced greater long-term potentiation recorded proximally than distally and the difference was significantly greater for proximal stimulation. When long-term potentiation was induced by stimulation of proximal and distal sites simultaneously, the difference between proximally- and distally-recorded long-term potentiation was significantly reduced. Paired-pulse facilitation was greater when recorded proximally but the variation in facilitation with stimulation and recording position was different from that obtained for long-term potentiation. Paired-pulse facilitation of response amplitude was slightly reduced after long-term potentiation, but the change did not depend on the stimulation-recording arrangement; facilitation of response slope was not reliably affected by long-term potentiation. These results indicate that the shape of the dendritic field excitatory postsynaptic potential is influenced by the relative position of recording electrodes in stratum radiatum; when the position is such that after-potentials are minimized, long-term potentiation produces a decrease in the decay time constant of the synaptic field potential.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 7477873 TI - Population trends in the fine spatial re-organization of synaptic elements in forebrain regions of chicks 0.5 and 24 hours after passive avoidance training. AB - Two regions in the forebrain of domestic chicks (Gallus domesticus), the intermediate and medial hyperstriatum ventrale and the lobus parolfactorius, have previously been shown to be important centres of biochemical, pharmacological and physiological change following one-trial passive avoidance training. The purpose of the present study was to examine, at the electron microscopic level, the fine spatial re-arrangement of synaptic structures in the intermediate and medial hyperstriatum ventrale (at 30 min), and in the lobus parolfactorius (at 24 h), post-training using comprehensive biometrical designs, image analysis and stochastic approaches. In intermediate and medial hyperstriatum ventrale, no significant differences in the numerical density of synapses either between control and trained chicks, or between hemispheres, were revealed using the disector method. However, after training, a nested-ANOVA demonstrated an increase in the thickness of pre- and post-synaptic electron densities (estimated via image analysis) only in the left intermediate and medial hyperstriatum ventrale, whereas synaptic apposition zone profiles increased in length bilaterally. In presynaptic terminals from the intermediate and medial hyperstriatum ventrale, stochastic analysis revealed that training resulted in the re-distribution of synaptic vesicles between two spatial pools relative to synaptic apposition zones, in both hemispheres producing a large number of synaptic vesicles closer to synaptic apposition zones; a nearest neighbour analysis of synaptic apposition zone profiles indicated that the lateral shape of the synaptic apposition zone after training is more complex in both hemispheres. In the lobus parolfactorius at 24 h post-training the main changes in synaptic fine structure involved a shift of synaptic vesicles away from synaptic apposition zones in the right hemisphere with the distance between synaptic apposition zones decreasing; in the left lobus parolfactorius, synaptic apposition zones became more regular/round in shape with a greater distance between them after training. These data suggest that the initial acquisition of memory involves population changes in the fine spatial organization of synaptic vesicles and synaptic apposition zones in synapses in the intermediate and medial hyperstriatum ventrale, which indicate a possible tendency towards greater synaptic efficacies. These changes are as dynamics as the molecular changes which have hitherto been considered the preserve of short-term correlates of memory formation. PMID- 7477874 TI - Altered synaptophysin expression as a marker of synaptic pathology in schizophrenia. AB - It has been proposed that synaptic density or synaptic innervation may be altered in schizophrenia as a correlate of the neurodevelopmental pathology of the disease. Synaptophysin is a synaptic vesicle protein whose distribution and abundance provides a synaptic marker which can be reliably measured in post mortem brain. We have used in situ hybridization histochemistry and immunoreactivity to assess the expression of synaptophysin messenger RNA and protein respectively in medial temporal lobe from seven schizophrenics and 13 controls. In the schizophrenic cases, synaptophysin messenger RNA was reduced bilaterally in CA4, CA3, subiculum and parahippocampal gyrus, with a similar trend in dentate gyrus but no change in CA1. It was also decreased in terms of grains per pyramidal neuron in the affected subfields. In parahippocampal gyrus, the loss of synaptophysin messenger RNA per neuron in schizophrenia was greater in deep than superficial laminae. A parallel study in rats showed no effect of haloperidol treatment upon hippocampal synaptophysin messenger RNA, suggesting that neuroleptic treatment does not underlie the reductions found in schizophrenia. In the right medial temporal lobe of schizophrenics, we confirmed the correlation of synaptophysin messenger RNA abundance between ipsilateral subfields seen in both hemispheres of control brains. However, these correlations were not observed in the left medial temporal lobe of the schizophrenic cases. Synaptophysin immunoreactivity in schizophrenia showed no significant differences in any subfield compared to controls. Our data support the broad hypothesis that synaptic pathology occurs in schizophrenia. In so far as synaptophysin expression is a marker for synaptic density, the data suggest that pyramidal neurons within the medial temporal lobe may form fewer synapses. However, the lack of any significant differences in synaptophysin immunoreactivity despite the loss of encoding messenger RNA means that this conclusion must be drawn cautiously. There are several plausible explanations for the preservation of synaptophysin immunoreactivity despite reductions in transcript abundance; one possibility is that the inferrred loss of synapses occurs in extra-hippocampal sites to which the affected pyramidal neurons project. For example, the reduction in synaptophysin messenger RNA in subicular neurons may be accompanied by decreased density of synaptic terminals in the nucleus accumbens. Such differences in the efferent synaptic connectivity of the hippocampus have previously been hypothesized to be an important component of the circuitry underlying schizophrenia. PMID- 7477875 TI - Mesopontine neurons in schizophrenia. AB - Findings reported here show that there is a significant increase in the number of neurons in the pedunculopontine nucleus in most schizophrenic patients compared to age-matched controls. Nicotinamide adenine dinucleotide phosphate diaphorase histochemistry was used to label putative cholinergic neurons in the pedunculopontine nucleus and laterodorsal tegmental nucleus, while noradrenergic locus coeruleus neurons were labeled immunocytochemically using an antibody to tryosine hydroxylase. Cell counts of these neuronal groups were carried out using a Biographics image analysis system. We found significantly increased cell numbers in the pedunculopontine nucleus of schizophrenic patients compared to controls. The number of laterodorsal tegmental nucleus neurons was increased but this was not statistically significant. However, the total cell counts for pedunculopontine and laterodorsal tegmental nuclei were significantly higher in schizophrenic subjects. The number of locus coeruleus noradrenergic neurons was similar in both groups. These results implicate the brainstem reticular formation as a pathophysiological site in at least some patients with schizophrenia. In addition, these findings suggest a developmental etiology for the disease and account for some, but not all, of the symptoms of schizophrenia, including sensory gating abnormalities, sleep-wake disturbances and, perhaps, hallucinations. Overdriving of thalamic and substantia nigra function by cholinergic afferents from the midbrain may account for some of the symptoms seen in schizophrenia. These findings suggest that, at least in some schizophrenic patients, there is an increased number of neurons in the cholinergic arm of the reticular activating system. This may explain some of the symptoms of schizophrenia and points to a prenatal disturbance as one of the possible causes of the disease. PMID- 7477876 TI - Antipsychotic drugs induce Fos protein in the thalamic paraventricular nucleus: a novel locus of antipsychotic drug action. AB - Monitoring expression of c-fos and other immediate-early genes has proven a useful method for determining potential sites of action of antipsychotic drugs. Most studies of the effects of antipsychotic drugs on immediate-early gene expression have focused on the basal ganglia and allied cortical regions. We now report that clozapine administration markedly increases both the number of cells expressing Fos protein-like immunoreactivity and the amount of Fos protein in the thalamic paraventricular nucleus, but not the contiguous mediodorsal thalamic nucleus. Comparable doses of several dopamine D2-like antagonists, including raclopride, sulpiride, remoxipride and haloperidol, did not induce Fos expression in the paraventricular nucleus. However, loxapine and very high doses of haloperidol resulted in a small but significant increase in paraventricular nucleus Fos expression. The dopamine D1 receptor antagonist SCH23390 did not induce Fos in the paraventricular nucleus or alter the magnitude of the clozapine elicited increase in Fos expression. The serotonergic 5-hydroxytryptamine2a/2c antagonist ritanserin, alone or in combination with sulpiride, did not increase Fos expression in the paraventricular nucleus. Similarly, the 5 hydroxytryptamine2:D2 antagonist risperidone did not change the amount of Fos protein in the paraventricular nucleus. Neither the alpha 1 adrenergic antagonist prazosin nor the muscarinic cholinergic antagonist scopolamine mimicked the effect of clozapine. The key placement of the paraventricular nucleus as an interface between the reticular formation and forebrain dopamine systems suggests that this thalamic nucleus may be an important part of an extended neural network subserving certain actions of antipsychotic drugs. PMID- 7477877 TI - Priming of a D1 dopamine receptor behavioural response is dissociated from striatal immediate-early gene activity. AB - Repeated administration of direct-acting (apomorphine, SKF-38393, quinpirole) or indirect-acting (amphetamine, cocaine) dopaminergic agonists can produce enhancement of locomotor and sterotypic behaviours in response to subsequent dopamine agonist challenge. This sensitization of dopamine receptors, known as priming or reverse tolerance, is long-lasting and appears to be dependent upon the participation of the N-methyl-D-asparate excitatory amino acid receptor. The mechanism underlying dopamine receptor sensitization is not understood. Mounting evidence suggests that immediate-early genes may provide a link whereby extracellular stimuli are converted into long-term changes in neuronal activity. In the present study, behavioural measurements and immunohistochemical techniques were used to determine whether induction of the immediate-early gene c-fos is critical to the mechanism underlying priming of a D1-mediated behavioural response. It was demonstrated that in drug-naive rats bearing unilateral 6 hydroxydopamine lesions of the dopaminergic nigrostriatal pathway, the mixed D1/D2 agonist apomorphine produced a dramatic increase in the expression of Fos like immunoreactivity in the ipsilateral caudoputamen, nucleus accumbens and globus pallidus, and was a potent primer of SKF-38393-mediated rotational behaviour. In contrast, saline administration did not increase Fos expression and did not prime SKF-38393-elicited rotation. Preadministration of MK-801 at 0.5 mg/kg significantly reduced apomorphine's effect on Fos expression and prevented apomorphine priming of SKF-38393-induced rotation. However, at a lower dose of 0.1 mg/kg, MK-801 had little effect on apomorphine-mediated Fos expression but did block the priming response. In another experiment, the D2 family-selective agonist quinpirole was found to be an affective primer of SKF-38393-mediated rotation, and to produce increase Fos expression in the ipsilateral globus pallidus only. Preadministration of MK-801 at 0.1 mg/kg blocked quinpirole priming of SKF-38393-mediated rotation and significantly reduced the number of Fos-positive neurons in the ipsilateral globus pallidus. Administration of the indirect dopamine agonist amphetamine increased Fos expression in the intact striatum, but not in the ipsilateral (lesioned) striatum or globus pallidus, and did not sensitize (prime) animals to behavioural effects of SKF-38393. In a separate group of animals. Northern blot analysis demonstrated that a priming dose of apomorphine significantly increased the messenger RNA signals for c-fos, c-jun, ngfi-A and jun-B in denervated striatum. Administration of 0.1 mg/kg MK 801 prior to apomorphine had no significant effect on signal intensities.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 7477878 TI - Preproenkephalin and preprotachykinin messenger RNA expression in normal human basal ganglia and in Parkinson's disease. AB - Striatal expression of preproenkephalin and preprotachykinin messenger RNA was studied in normal controls and in patients with Parkinson's disease using in situ hybridization histochemistry. In controls, preproenkephalin messenger RNA was expressed in a population of medium-sized neurons of mean cross-sectional area 165 microns 2, accounting for 66% of striatal medium-sized neurons, whereas preprotachykinin messenger RNA was expressed in a population of medium-sized neurons of mean cross-sectional area 204 microns 2 (23% larger than those expressing enkephalin, P < 0.05), accounting for 58% of medium-sized striatal neurons. Much lower levels of both preproenkephalin messenger RNA and preprotachykinin messenger RNA were expressed by large neurons in the globus pallidus and substantia nigra reticulata. In addition, preproenkephalin messenger RNA was expressed at low levels by neurons in the subthalamic nucleus. In Parkinson's disease cases, there was a statistically significant increase in preproenkephalin messenger RNA expression in the body of the caudate (109% increase, P < 0.05) and in the intermediolateral putamen (55% increase, P < 0.05) due to an increase in the level of gene expression per neuron rather than an increase in the number of neurons expressing preproenkephalin messenger RNA. Similar increases were observed in other putaminal subregions and in the putamen as a whole, but these did not reach statistical significance. No change in preprotachykinin messenger RNA expression was detected. These findings demonstrate selective up-regulation of a striatal neuropeptide system in Parkinson's disease compatible with increased activity of the "indirect" striatopallidal pathway, which is thought to play a crucial role in the pathophysiology of akinesia and rigidity in this condition. PMID- 7477879 TI - Hemodynamic regulation of tyrosine hydroxylase messenger RNA in medullary catecholamine neurons: a c-fos-guided hybridization histochemical study. AB - Medullary catecholamine cell groups are involved in multiple modes of cardiovascular regulation and display indices of functional activation, including widespread c-fos expression, in response to hypotensive hemorrhage. Assessments of the impact of such challenges on transmitter-related gene expression are complicated by the biochemical and connectional heterogeneity that characterize these cell groups. Quantitative hybridization histochemical methods were used to follow the effects of 15% hemorrhage on levels of messenger RNA encoding tyrosine hydroxylase, the rate-limiting enzyme in catecholamine biosynthesis, in medullary aminergic neurons; concurrent staining for nuclear Fos-immunoreactivity permitted comparisons between cells that ostensibly were and were not targeted by the challenge. Increased levels of tyrosine hydroxylase messenger RNA were detected in Fos-immunoreactive neurons in all cell groups examined. Mean maximal increases ranged between 133 and 192% of control values, and were attained within 0.5-1 h post-hemorrhage in noradrenergic (A1 and A2) cell groups, and at 2 h in adrenergic ones (C1, C2, and C2d or dorsal strip). By 4 h after the challenge, tyrosine hydroxylase messenger RNA levels in Fos-immunoreactive neurons in all cell groups had returned to control values. By contrast, tyrosine hydroxylase messenger RNA in non-Fos-immunoreactive cells either did not change significantly over the course of the experiment (C2 and C2d), or showed a rapid and transient increase, whose magnitude tended to be less than that seen in Fos-immunoreactive cells. c-fos messenger RNA was prominently induced in catecholaminergic neurons in each of the medullary cell groups examined at 0.5 h after hemorrhage, suggesting that the early tyrosine hydroxylase messenger RNA response to hemorrhage in non-Fos-immunoreactive cells preceded the capacity of responsive neurons to manifest detectable Fos protein expression. These findings indicate that hemorrhage up-regulates tyrosine hydroxylase messenger RNA levels in medullary catecholaminergic cell groups which have access to adaptive neuroendocrine and/or autonomic control systems. The approach employed here should prove of general utility in assessing the impact of environmental events on messenger RNA expression in connectionally heterogeneous cell groups that share a common biochemical phenotype. PMID- 7477880 TI - Clonidine and rilmenidine suppress hypotension-induced Fos expression in the lower brainstem of the conscious rabbit. AB - Our current knowledge of the sites of action of the centrally-acting antihypertensive drug clonidine is based almost entirely on experiments in anesthetized animals. The aim of this study was to determine, in conscious rabbits, the sites of action in the brainstem of systemically administered clonidine, as well as its oxazoline analog rilmenidine. Three groups of experiments were carried out. In the first group, hypotension was produced by continuous intravenous infusion of sodium nitroprusside, at a rate sufficient to decrease arterial pressure by 20-30 mmHg, maintained for a period of 60 min. In the second and third groups of experiments, sustained hypotension was also produced by nitroprusside infusion as in the first group, but this was preceded by intravenous injection of clonidine (7-30 micrograms/kg i.v.) or rilmenidine (150-300 micrograms/kg i.v.), respectively. In confirmation of our previous study [Li Y.-W. and Dampney R. A. L. (1994) Neuroscience 61, 613-634], hypotension produced by nitroprusside alone induced a large increase (compared to sham control experiments) in the neuronal expression of Fos (a marker of neuronal activation) in the nucleus of the solitary tract, area postrema, the rostral, intermediate and caudal parts of the ventrolateral medulla, A5 area, locus coeruleus and subcoeruleus, and parabrachial nucleus. In comparison with this group, in rabbits pretreated with clonidine the numbers of Fos-positive cells were greatly reduced (by 76-94%) in the rostral, intermediate and caudal parts of the ventrolateral medulla, area postrema, A5 area, locus coeruleus and subcoeruleus. Clonidine pretreatment also caused a more moderate reduction (by 45%) in the number of Fos-positive cells in the nucleus of the solitary tract, but had no effect on Fos expression in the parabrachial nucleus. Double-labeling for tyrosine hydroxylase and Fos immunoreactivity showed that clonidine pretreatment greatly reduced the numbers of both catecholamine and non catecholamine Fos-positive neurons. Rilmenidine pretreatment also greatly reduced Fos expression in the lower brainstem, with a very similar pattern to that observed after clonidine pretreatment. The results indicate that in conscious animals both clonidine and rilmenidine cause a widespread but selective inhibition of neurons in the pons and medulla that are normally activated by a hypotensive stimulus. In contrast to previous observations in anesthetized animals, the results suggest that (i) systemic administration of both drugs inhibits non-catecholamine as well as catecholamine neurons in the ventrolateral medulla, and (ii) the regional pattern of neuronal inhibition following administration of equipotent hypotensive doses of both drugs is very similar. PMID- 7477881 TI - Involvement of the noradrenergic afferents from the nucleus tractus solitarii to the supraoptic nucleus in oxytocin release after peripheral cholecystokinin octapeptide in the rat. AB - Activation of abdominal vagal afferents by peripheral injection of cholecystokinin octapeptide induces oxytocin release into the circulation. To test the hypothesis that cholecystokinin increases oxytocin release via activation of noradrenergic afferents from the brainstem, we injected rats with 5 amino-2,4-dihydroxy-alpha-methylphenylethylamine, a selective neurotoxin to noradrenergic fibres, into a lateral cerebral ventricle. The neurotoxin treatment reduced the noradrenaline content in the hypothalamus by 75% and reduced the oxytocin secretion in response to cholecystokinin by over 90%. In separate experiments, the neurotoxin was injected unilaterally in the vicinity of the supraoptic nucleus to test whether direct noradrenergic afferents to the supraoptic nucleus are involved in the response to cholecystokinin. The injection reduced the immunoreactivity for dopamine beta-hydroxylase in the supraoptic nucleus and significantly decreased the number of the supraoptic neurons expressing Fos-like protein after cholecystokinin but not after hypertonic saline. In further experiments, rhodamine-conjugated latex microspheres were injected into the supraoptic nucleus to retrogradely label afferent neurons, and the brains were processed with double-immunohistochemistry for tyrosine hydroxylase and Fos-like protein. In the C2/A2 but not the C1/A1 region of the brainstem, cholecystokinin increased the expression of Fos-like protein in the population of retrogradely-labelled catecholaminergic cells. In the C2/A2 region, the majority of retrogradely labelled cells expressing Fos-like protein after cholecystokinin were catecholaminergic. We conclude that noradrenergic afferents from the A2 but not from the A1 region of the brainstem to the hypothalamus mediate, at least in part, oxytocin release following cholecystokinin. PMID- 7477882 TI - Mesencephalic grafts partially restore normal nigral dynorphin levels in 6 hydroxydopamine-lesioned rats treated chronically with L-dihydroxyphenylalanine. AB - An increase of dynorphin levels is commonly observed in the substantia nigra of 6 hydroxydopamine-lesioned rats chronically treated with daily injections of L DOPA. This study investigates the potential of fetal mesencephalic grafts to restore normal levels of dynorphin in such cases. After 19 consecutive days of treatment with L-DOPA, lesioned rats with the most severe nigral cell loss showed increased levels of dynorphin in the substantia nigra ipsilateral to the lesion, as expected. The changes were assessed by standard immunohistochemical techniques combined with the use of an image analysis system. Such changes were not observed in the substantia nigra of rats that received fetal mesencephalic cells in the striatum six months prior to the beginning of the chronic treatment. However, only animals displaying heavy loss of dopaminergic neurons in the substantia nigra pars compacta showed significant changes of dynorphin levels in the substantia nigra following drug treatment. Our results show that fetal nigral cells transplanted into the striatum have the potential to prevent biochemical changes observed in the basal ganglia induced by the lesion of the nigrostriatal pathway and chronic treatment with L-DOPA. It is still hypothesized from studies in rodents that this peptide may play a role in the appearance of DOPA-induced dyskinesia, because dynorphin levels increase in the substantia nigra pars reticulata after L-DOPA treatment. If this happens to be the case, then the use of fetal nigral grafts could therefore be an important step to prevent the induction of dyskinesia after chronic L-DOPA treatment. PMID- 7477883 TI - Peripheral nociceptive effects of alpha 2-adrenergic receptor agonists in the rat. AB - We have previously shown that norepinephrine can produce hyperalgesia via an alpha 2-adrenergic receptor mechanism. The alpha 2-adrenergic receptor agonist clonidine has, however, also been shown to produce peripheral analgesia. In view of the multiple alpha 2-subtypes currently known (i.e. alpha 2A, alpha 2B and alpha 2C), we evaluate the alpha 2-receptor subtypes mediating norepinephrine induced peripheral hyperalgesia and clonidine analgesia. Norepinephrine and the alpha 2-adrenergic agonists clonidine and UK 14,304 (1-1000 ng), when co-injected with the calcium ionophore A23187 (1000 ng) produced dose-dependent hyperalgesia in the Randall-Selitto paw withdrawal test. Norepinephrine (100 ng) hyperalgesia was dose-dependently antagonized by alpha 2-adrenergic receptor antagonists. From the estimated ID50, the rank order of potency was: SK&F 104856 (alpha 2B) approximately imiloxan (alpha 2B) > rauwolscine (alpha 2C) >> BRL 44408 (alpha 2A). Norepinephrine hyperalgesia was not significantly affected by pertussis toxin treatment. Prostaglandin E2 (100 ng) hyperalgesia was inhibited dose dependently, by clonidine and UK 14,304. Rauwolscine was more potent in reversing the inhibitory effect of clonidine on prostaglandin E2 than imiloxan while BRL 44408 was ineffective. The inhibitory effect of clonidine on prostaglandin E2 hyperalgesia was reversed by pertussis toxin. These data suggest that alpha 2B subtype receptors mediate (norepinephrine hyperalgesia while the antinociceptive effect of alpha 2-agonist is mediated by the alpha 2C-subtype receptor. Differential coupling of these receptor subtypes to second messenger systems and location on different cell types in the rat paw may explain, at least in part, their differential responses to alpha 2-agonist stimulation, leading to hyperalgesia and analgesia. PMID- 7477884 TI - Mechanical stimulation increases intracellular calcium concentration in nodose sensory neurons. AB - The cellular mechanisms involved in activation of mechanosensitive visceral sensory nerves are poorly understood. The major goal of this study was to determine the effect of mechanical stimulation on intracellular calcium concentration ([Ca2+]i) using nodose sensory neurons grown in culture. Primary cultures of nodose sensory neurons were prepared by enzymatic dispersion from nodose ganglia of 4-8 week old Sprague-Dawley rats. Whole cell [Ca2+]i was measured by a microscopic digital image analysis system in fura-2 loaded single neurons. Brief mechanical stimulation of individual nodose sensory neurons was achieved by deformation of the cell surface with a glass micropipette. In 31 of 50 neurons (62%), mechanical stimulation increased [Ca2+]i from 125 +/- 8 to 763 +/- 89 nM measured approximately 10 s after stimulation. [Ca2+]i then declined gradually, returning to near basal levels over a period of minutes. [Ca2+]i failed to increase after mechanical stimulation in the remaining 19 neurons. The mechanically-induced rise in [Ca2+]i was essentially abolished after the neurons were incubated for 5-10 min in zero Ca2+ buffer (n = 7) or after addition of gadolinium (10 microM), a blocker of stretch-activated ion channels (n = 5). The effect of gadolinium was reversed after removal of gadolinium. The results indicate that: (1) mechanical stretch increases [Ca2+]i in a subpopulation of nodose sensory neurons in culture, and (2) the stretch-induced increase in [Ca2+]i is dependent on influx of Ca2+ from extracellular fluid and is reversibly blocked by gadolinium. The findings suggest that opening of stretch-activated ion channels in response to mechanical deformation leads to an increase in Ca2+ concentration in visceral sensory neurons.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7477885 TI - Plasticity in the synthesis and storage of substance P and calcitonin gene related peptide in primary afferent neurons during peripheral inflammation. AB - Several indices of peptidergic, primary afferent neural transmission in rat at the level of the lumbar spinal cord exhibited differential changes over time in response to adjuvant-induced inflammation of the hindpaw. The indices were measurements of the production of messenger RNA encoding the precursors for substance P and calcitonin gene-related peptide in dorsal root ganglia, the storage of substance P and calcitonin gene-related peptide in the dorsal spinal cord and the release of the peptides evoked by application of capsaicin to the dorsal spinal cord. A 47% decrease in the content of immunoreactive substance P in the dorsal half of the lumbar spinal cord, as determined by radioimmunoassay, was measured at 6 h following the injection of complete Freund's adjuvant into the hindpaw. Decreased content of immunoreactive SP persisted for four days, but was no longer present at eight days after the adjuvant injection. The content of immunoreactive calcitonin gene-related peptide in the dorsal spinal cord was decreased by 29% at one day following the injection of adjuvant into the rat hindpaw and 43% at two days; the content then increased to a level greater than that of control animals at eight days. The amount of messenger RNA encoding preprotachykinin and prepro-calcitonin gene-related peptide in L4-L6 dorsal root ganglia was determined from northern blot analysis of the total messenger RNA extracted from the dorsal root ganglia. Each species of messenger RNA had increased compared to the control animals at two days following the injection of adjuvant into the rat hindpaws and remained elevated after eight days. Thus, an increase in the messenger RNAs encoding substance P and calcitonin gene-related peptide in the dorsal root ganglia preceeded the recovery of the content of the peptides in the spinal cord. Morphometric studies of calcitonin gene-related peptide-immunoreactive perikarya in the L4 dorsal root ganglia indicated that the increase in messenger RNA occurred in neurons of the size that normally express calcitonin gene-related protein. Radioimmunoassay of the superfusate of the dorsal half of the lumbar spinal cord was used to measure the release of immunoreactive substance P and immunoreactive calcitonin gene-related protein in vitro. Although the basal release of immunoreactive substance P and immunoreactive calcitonin-gene related protein from the dorsal spinal cord was constant throughout the time points examined, changes occurred in the release of peptide evoked by 10 microM capsaicin. The capsaicin-evoked release of immunoreactive substance P was decreased at 6 h and eight days post-injection of adjuvant.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 7477886 TI - Cyclic AMP mediates the prostaglandin E2-induced potentiation of bradykinin excitation in rat sensory neurons. AB - Prostaglandins enhance the sensitivity of sensory neurons to excitatory chemical agents such as bradykinin. The intracellular transduction cascades mediating this potentiation remain largely unknown. We have examined the role of cyclic AMP in the prostaglandin E2-induced potentiation of sensory neurons. Pretreatment with agents that elevate intracellular cyclic AMP levels enhances the number of action potentials elicited by bradykinin in a manner analogous to that of prostaglandin E2. The prostaglandin E2-induced potentiation of the number of bradykinin elicited action potentials is blocked by either inhibition of adenylyl cyclase or protein kinase A. Therefore, our results suggest that prostaglandin E2 activates adenylyl cyclase to increase intracellular cyclic AMP, which in turn activates protein kinase A. Presumably activation of protein kinase A leads to increased levels of protein phosphorylation that then contribute to the enhancement of neuronal sensitivity to excitatory chemical agents. PMID- 7477887 TI - Prenatal corticosterone increases spontaneous and d-amphetamine induced locomotor activity and brain dopamine metabolism in prepubertal male and female rats. AB - Recently, both glucocorticoid receptor immunoreactivity and glucocorticoid receptor messenger RNA levels were found in multiple brain areas, especially in the neuroepithelium during the late prenatal development of the rat brain. To better understand the potential influence of stress on fetal brain development by release of maternal adrenocortical steroids, we have investigated the effects of corticosterone administration to pregnant rats on the locomotor activity of their prepubertal offspring. On day 16 of pregnancy female rats were implanted with either placebo or corticosterone pellets (release of 2.4 mg/day for seven days). After birth their offspring were nursed by foster mothers to avoid any postnatal effects of the corticosterone pellets. At three weeks of age, the offspring were tested for spontaneous motor behaviours. Both male and female offspring from corticosterone treated mothers showed significantly increased spontaneous ambulation, motility and rearing compared to placebo treated groups. No significant sex differences were found in locomotor activity between male and female offspring from placebo groups. Following d-amphetamine (1.5 mg/kg) treatment, a preferential dopamine releasing agent, we observed a significant increase in ambulation, motility and rearing activity in the male offspring treated with corticosterone. In the female offspring, only the rearing activity was significantly higher after d-amphetamine treatment in the prenatal corticosterone group compared with the placebo treated group. Basal dopamine metabolism (dihydroxyphenylacetic acid/dopamine ratio) was increased in the dorsal striatum and ventral striatum of male and female offspring from corticosterone-treated dams. In the male offspring, corticosterone treatment was associated with a disappearance of the right side dominance of dopamine metabolism in the dorsal striatum.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7477888 TI - Different neuronal and glial cell groups in corticomedial amygdala react differently to neonatally administered estrogen. AB - The percentage of labeled neurons and glial cells in the phylogenetically older corticomedial part of the amygdala was investigated in control and estrogen treated rats using [3H]thymidine autoradiography. Newborn, three-day-old female and male Wistar rats were treated with a single dose of 1 mg of estrogen and killed at the age of 10 days. The percentage of labeled neuronal and glial cells was determined by stereological methods in the medial, cortical and central nuclei, respectively. In treated male rats, the percentage of labeled neurons and glial cells in these nuclei was significantly greater on the 10th day of life as compared to controls, except for glial cells in the nucleus centralis. In treated female rats, the percentage of labeled neurons in all three nuclei was comparable to controls, while the percentage of labeled glial cells was increased in the nucleus medialis, decreased in the nucleus centralis and unchanged in the nucleus corticalis. Our results indicate clear sex- and region-specific differences in the reactivity of both neurons and glia to neonatally administered estrogen. PMID- 7477889 TI - Distribution of glutamate receptor subtypes in the vertebrate retina. AB - The distribution of glutamate receptor subunit/subtypes in the vertebrate retina was investigated by immunocytochemistry using anti-peptide antibodies against AMPA (GluR1-4), kainate (GluR6/7) and metabotropic (mGluR1 alpha) receptors. All receptor subtypes examined are present in the mammalian retina, but they are distributed differentially. GluR1 is present in the inner plexiform layer as well as amacrine and ganglion cell bodies. GluR2 is present mainly in the outer plexiform layer and bipolar cells. An anti-GluR2/3 antibody labels both plexiform layers and various cell bodies in the inner nuclear layer and the ganglion cell layer. GluR4 is present on Muller glial cells. In the goldfish retina, GluR2 immunoreactivity is prominent in the Mb type of ON-bipolar cells, including the dendrites and the large synaptic terminal. The putative dendritic localization is surprising, because no depolarizing conductance increase induced by glutamate is thought to be present in these cells. An AMPA receptor at a presynaptic terminal is also unusual, and probably provides feedback control of glutamate release. GluR6/7 is most widespread in the retina, being present in horizontal, bipolar, amacrine and ganglion cells. Ion channels composed of GluR6 are now known to be phosphorylated by protein kinase A, resulting in current potentiation. This property and our present observation together suggest that the glutamate receptors previously studied electrophysiologically by others in horizontal cells may contain GluR6. mGluR1 alpha is found mostly in the inner plexiform layer; its localization partially overlaps with that of the inositol trisphosphate receptor in the retina. Our results suggest that, in the retina, glutamate receptor subtypes may be expressed in selective cell types according to their specific functions. PMID- 7477890 TI - [D-Pen2,D-Pen5]enkephalin, a delta opioid agonist reduces endogenous aluminum content in the rat central nervous system. AB - The in vivo effects of [D-Pen2,D-Pen5]enkephalin, a cyclic peptide agonist with high affinity and selectivity for the delta opioid receptors, on the endogenous aluminum content of selected areas of rat brain and spinal cord were studied by means of atomic absorption spectrophotometry. Intracerebroventricular injection of a subanalgesic dose of [D-Pen2,D-Pen5]enkephalin (0.2 microgram/3 microliters) produced a transient, time-dependent reduction of the aluminum content. This effect was statistically significant in the frontal cortex, hippocampus and striatum, but did not reach the level of significance in the medulla and thoracic spinal cord. The partial depleting effect of [D-Pen2,D-Pen5]enkephalin on aluminum content, in the range of 0.2-1.0 micrograms/3 microliters, was dose dependent and could be reversed by naloxone pretreatment. Serum aluminum levels were unchanged after [D-Pen2,D-Pen5]enkephalin treatment. Chronic (five weeks), systemic AlCl3 treatment increased the endogenous aluminum content in all central nervous system areas examined. Interestingly, [D-Pen2,D-Pen5]enkephalin i.c.v. produced a slight depletion of this elevated metal level in these areas to values not significantly different from those of the respective control values. Chronic in vivo, as well as in vitro, effects of aluminum on opioid receptor binding characteristics were also studied. Neither the specific binding of [3H][D-Pen2,D Pen5]enkephalin nor [3H]Tyr-D-Ala-Gly-NMePhe-Gly-ol to membranes of frontal or parietal cortices, striatum or hippocampus, prepared from rats chronically treated with AlCl3, were affected.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7477891 TI - Increased expression of neuronal nitric oxide synthase in dorsal root ganglion neurons after systemic capsaicin administration. AB - Nitric oxide synthase which is constitutively expressed in some neurons can be induced in other neurons by pathological conditions. For example, sciatic nerve or pelvic nerve transection induced nitric oxide synthase expression in lumbosacral dorsal root ganglion cells. This occurred in small to medium sized dorsal root ganglion neurons suggesting that this change might be limited to, or most prominent in, C-fiber afferents. These afferents are also very sensitive to the neurotoxin, capsaicin, which can deplete neuropeptide stores in C-fibers and in high doses cause degeneration of C-fiber afferent pathways. The present study was undertaken to determine if a chemically induced injury elicited in small diameter afferent neurons by systemic administration of capsaicin can induce a change in nitric oxide synthase expression similar to that elicited by peripheral nerve injury. Following capsaicin pretreatment (three days) a significant increase in the number of nitric oxide synthase-immunoreactive cells was detected in dorsal root ganglia. The increase was most prominent (34-63 fold) in the L5-S1 dorsal root ganglia but also substantial (2-17 fold) in cervical, caudal thoracic and rostral lumbar dorsal root ganglia. These results indicate that the expression of nitric oxide synthase-immunoreactivity in afferent neurons in the dorsal root ganglia is plastic and can be upregulated in response to chemical stimulation and/or injury. It is possible that nitric oxide formed by enhanced expression of nitric oxide synthase may play a role in capsaicin-induced neurotoxicity. PMID- 7477892 TI - Fos expression in neurons projecting to the pressor region in the rostral ventrolateral medulla after sustained hypertension in conscious rabbits. AB - Previous studies in anaesthetized animals have shown that the baroreflex control of sympathetic vasomotor activity is mediated to a large extent by inhibitory inputs to sympathoexcitatory pressor neurons in the rostral part of the ventrolateral medulla. The aim of this study was to determine, in conscious rabbits, the distribution of neurons within the brain that have two properties characteristic of interneurons conveying baroreceptor signals to the rostral ventrolateral medulla: (i) they are activated by an increase in arterial pressure; and (ii) they project specifically to the rostral ventrolateral medulla pressor region. In a preliminary operation, an injection of the retrogradely transported tracer, fluorescent-labelled microspheres, was made into the physiologically identified pressor region in the rostral ventrolateral medulla. After a waiting period of one to eight weeks, hypertension was produced in the conscious rabbit by continuous intravenous infusion of phenylephrine at a rate sufficient to increase arterial pressure by approximately 20 mmHg, maintained for a period of 60 min. A control group of animals was infused with the vehicle solution alone. In confirmation of our previous study, hypertension produced by phenylephrine resulted in the neuronal expression of Fos (a marker of neuronal activation) in the nucleus of the solitary tract, area postrema, the intermediate and caudal parts of the ventrolateral medulla parabrachial complex, and in the central nucleus of the amygdala. Approximately 50% of the Fos-immunoreactive neurons in both the caudal and intermediate parts of the ventrolateral medulla were also retrogradely labelled from the rostral ventrolateral medulla pressor region; such double-labelled neurons were confined to a discrete longitudinal column located just ventrolateral to the nucleus ambiguus. Significant numbers of double-labelled neurons were also found in the nucleus of the solitary tract and area postrema, although these represented a much lower proportion (13-16%) of the total number of Fos-immunoreactive neurons in these regions. In the parabrachial complex, Fos-immunoreactive and retrogradely labelled neurons were largely separate populations, while in the amygdala they were entirely separate populations. In the control group of rabbits, virtually no double-labelled neurons were found in any of these regions. The results indicate that putative baroreceptor interneurons that project to the pressor region of the rostral ventrolateral medulla are virtually confined to the lower brainstem. In particular, they support the results of previous studies in anaesthetized animals indicating that neurons in the intermediate and caudal ventrolateral medulla convey baroreceptor signals to the rostral ventrolateral medulla pressor region, and extend them by demonstrating the precise anatomical distribution of these neurons.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 7477893 TI - Changes in the pattern of glutamate-like immunoreactivity in rat superior colliculus following retinal and visual cortical lesions. AB - We have investigated the pattern of glutamate-like immunoreactivity in the superficial layers of the rat superior colliculus by means of postembedding immunocytochemical methods for light and electron microscopy. At the light microscopic level, labelling was faintly to moderately intense in most perikarya of the stratum zonale, stratum griseum superficiale and stratum opticum. Furthermore, strong glutamate-immunoreactive terminal-like elements were accumulated most densely in stratum zonale, stratum griseum superficiale and stratum opticum. At the electron microscopic level, a postembedding immunogold method revealed that the vast majority of those labelled elements corresponded to retinal and visual cortical terminals. These profiles were about twice as heavily labelled as their postsynaptic partners. To determine the contribution of retinal and cortical afferents to the pattern of glutamate-like immunoreactivity, rats were subjected to right retinal ablation, left cortical ablation or combined right retinal and left cortical ablations. After retinal ablation, strongly labelled perikarya were observed in the retinorecipient layers. Furthermore, a prominent loss of glutamate-immunoreactive terminal-like elements occurred in stratum zonale and stratum griseum superficiale. Ipsilateral superior colliculus to cortical ablation exhibited subtle changes characterized by a moderate increase in perikaryal immunostaining in stratum zonale, stratum griseum superficiale and stratum opticum and by an apparent discrete reduction of labelled dots in stratum griseum superficiale and stratum opticum. In cases with combined lesions, strongly immunoreactive cell bodies and dendrites were accompanied by a massive disappearance of labelled terminal-like elements in stratum zonale, stratum griseum superficiale and stratum opticum. The effect of retinal and visual cortical ablations on the pattern of glutamate-like immunoreactivity suggests that these afferents are the major sources for glutamate-immunoreactive terminals in the rat superior colliculus. In addition, these findings provide further evidence for glutamate as neurotransmitter in the visual pathways studied. PMID- 7477894 TI - Regional suppression by lesions in the anterior third ventricle of c-fos expression induced by either angiotensin II or hypertonic saline. AB - Angiotensin II (250 pmol) infused into the cerebral ventricles of male rats induces the expression of c-fos in the subfornical organ, supraoptic and paraventricular nuclei of the hypothalamus, as well as in the lateral parabrachial nucleus, locus coeruleus and the nucleus of the solitary tract in the brainstem. Electrolytic lesions of the anteroventral third ventricle, principally the subcommissural (ventral) median preoptic nucleus, inhibited the dipsogenic response to i.c.v. angiotensin II and also suppressed c-fos expression in supraoptic nucleus, paraventricular nucleus, lateral parabrachial nucleus, locus coeruleus and nucleus of the solitary tract but not in the subfornical organ or dorsal median preoptic nucleus. The stimulating effect of i.c.v. angiotensin II on corticosterone was also reduced. Median preoptic nucleus lesions also suppressed the expression of c-fos following i.v. infusions of 6 micrograms angiotensin II in supraoptic nucleus and paraventricular nucleus but not in subfornical organ, dorsal median preoptic nucleus, lateral parabrachial nucleus, locus coeruleus and nucleus of the solitary tract. Median preoptic nucleus lesions reduced the dipsogenic effects of an intragastric infusion of hypertonic (1.5 M) saline and suppressed c-fos expression in supraoptic nucleus and paraventricular nucleus compared to sham-lesioned rats. However, c-fos expression was unaltered in subfornical organ, dorsal median preoptic nucleus lesions had no effect on the increased corticosterone induced by hypertonic saline. Subfornical organ lesions did not alter dipsogenic responses to i.c.v. angiotensin II, nor was the i.c.v. angiotensin II-induced expression of c-fos suppressed in the basal forebrain. These experiments show that the ventral median preoptic nucleus (but not the subfornical organ), part of the anteroventral third ventricle, is critical for the expression of c-fos in more caudal areas of the brain following i.c.v. angiotensin II. c-fos expression in supraoptic nucleus and paraventricular nucleus following i.v. angiotensin II is also dependent on an intact median preoptic nucleus, suggesting that supraoptic nucleus and paraventricular nucleus activation may be dependent on the median preoptic nucleus, and that suppression following i.c.v. infusions is not due to mechanical obstruction to infused peptide. However, there is a clear separation of the effects of i.c.v. and i.v. angiotensin II on brainstem structures. The median preoptic nucleus (but not the subfornical organ) seems essential for activation following the former but not the latter, suggesting alternative mechanisms for the effect of i.v. angiotension II on the brainstem.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 7477895 TI - Cyclic GMP-dependent protein kinase activation mediates inhibition of catecholamines secretion and Ca2+ influx in bovine chromaffin cells. AB - The effects of the membrane-permeable cGMP analogue, 8-bromoguanosine 3':5' cyclic monophosphate on acetylcholine-evoked catecholamine secretion and cytosolic calcium increases were studied in chromaffin cells from the bovine adrenal gland. Preincubation with 100 microM 8-bromoguanosine 3':5'-cyclic monophosphate during 10 and 30 min decreased the acetylcholine-evoked catecholamine release by 16 +/- 3% and 27 /+- 5%, respectively. The cytosolic calcium increases triggered by acetylcholine and 30 mM KCl were also inhibited by 30 min of preincubation with this compound by 27 +/- 4 and 34 /+- 12%, respectively. Changes in membrane potential induced by acetylcholine and KCl were not affected by preincubation with 8-bromoguanosine 3':5'-cyclic monophosphate. The cyclic GMP-dependent protein kinase inhibitor N-[2-(methylamino)ethyl]-5 isoquinoline sulfonamide dihydrochloride-at l micron abolished the inhibitory effect of 8-bromoguanosine 3':5'-cyclic monophosphate on acetylcholine-evoked calcium increase. By contrast, a potent and selective inhibitor against cyclic AMP-dependent protein kinase, N-[2-(p-bromocinnamylamino)ethyl]-5 isoquinolinesulfonamide did not block the 8-bromoguanosine 3':5'-cyclic monophosphate effect. Additionally, 8-bromoguanosine 3':5'-cyclic monophosphate stimulated histone F2b phosphorylation by a partial purified cGMP-dependent protein kinase from chromaffin cells. The extent of histone phosphorylation was reduced by N-[2-(methylamino)ethyl]-5-isoquinolinesulfonamide dihydrochloride and 8-(4-chlorophenylthio)-guanosine 3':5'-cyclic monophosphorothioate, Rp-isomer, a specific inhibitor against cyclic GMP-dependent protein kinase, whereas it was not modified by N-[2-(p-bromocinnamylamino)ethyl]-5-isoquinoline sulfonamide. The results suggest that the inhibitory effects of 8-bromoguanosine 3':5' cyclic monophosphate on chromaffin cells are mediated through the activation of cGMP dependent protein kinase.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7477896 TI - A1-receptor-mediated effect of adenosine on the release of acetylcholine from the myenteric plexus: role and localization of ecto-ATPase and 5'-nucleotidase. AB - No attempt has been made so far to classify the subtypes of presynaptic inhibitory adenosine receptors located in the myenteric plexus and to localize ecto-ATPase and 5'-nucleotidase in the intestine. The release of [3H]acetylcholine and smooth muscle responses to acetylcholine were measured and the effect of selective adenosine receptor ligands was studied using field stimulated isolated longitudinal muscle strips of guinea-pig ileum. Release of ATP and its hydrolysis rate were also measured using the luciferin-luciferase technique. A histochemical method combined with electron microscopy was used for localization of ecto-ATPase and 5'-nucleotidase, enzymes responsible for destruction of extracellular ATP, ADP and AMP. Subtype-selective A1-receptor agonists and antagonists inhibited and enhanced, respectively, the release of acetylcholine associated with neuronal activity. A significant amount of ATP was released in response to electrical stimulation and administration of carbamylcholine. The release of ATP was inhibited by atropine and 4 diphenylacetoxy-N-methylpiperidine methiodide, an M3-receptor antagonist. Hydrolysis of ATP was rapid and resulted in an accumulation of extracellular adenosine involved in presynaptic A1-receptor-mediated inhibition of acetylcholine release. While the inhibitory effect of adenosine and ATP was significantly potentiated by dipyridamol, an adenosine uptake blocker, that of 2 ms ATP was not. The effect of ATP was not competitively antagonized by 8 cyclopentyl-1,3-dipropylxanthine, a selective A1-receptor antagonist. In conclusion, axon terminals of cholinergic interneurons are equipped with inhibitory A1- and P2 gamma-receptors. Therefore, both adenosine and ATP control the release of acetylcholine through these receptors. ATP is mainly released from the smooth muscle in response to stimulation of M3-muscarinic receptors by endogenous acetylcholine (cascade transmission [Vizi E. S. et al. (1992) Neuroscience 50, 455-465]) and is rapidly hydrolysed by ecto-ATPase localized on the surface of the smooth muscle and axon terminals producing ADP and AMP, and by 5'-nucleotidase present only on the surface of smooth muscle cells producing adenosine. PMID- 7477897 TI - Physiological and regenerative acetylcholine release from motor nerve: differential inhibitions by vesamicol and omega-agatoxin IVA. AB - Stimulation of mammalian motor neurons can elicit Ca(2+)-dependent regenerative release of acetylcholine and prolonged endplate depolarization when the enzymatic degradation of the neurotransmitter is inhibited. Unlike physiological phasic release of acetylcholine, the regenerative release is sensitive to L-type Ca2+ channel blockers. We studied the effects of vesamicol (an inhibitor of active transport of acetylcholine into synaptic vesicles) and omega-agatoxin IVA (a blocker of the motor nerve P-type Ca2+ channel) on these two types of acetylcholine release to compare the vesicle pools and Ca2+ channels responsible for the release. When coupled with repetitive stimulations, vesamicol decreased mean amplitude of miniature endplate potentials, resulting in a skewed distribution to lower amplitude, reduced quantal content of endplate potentials and decreased immediate available pool of acetylcholine. omega-Agatoxin IVA had no effect on miniature endplate potential but inhibited quantal content of endplate potential. The mean inhibitory concentration was around 5-10 nM. Vesamicol and omega-agatoxin IVA decreased the probability of triggering regenerative release. However, the magnitude and duration of regenerative release, once triggered, were not depressed by either agent. It appears that the majority of Ca2+ necessary for regenerative release is translocated via omega agatoxin IVA-insensitive Ca2+ channels, which can be activated by prolonged depolarization of nerve terminals induced by accumulated acetylcholine. The results suggest that different Ca2+ channels are activated in the regenerative (L type) and phasic (P-type) acetylcholine release, which utilize different pools of synaptic vesicles. PMID- 7477898 TI - Co-expression of AMPA/kainate receptor-operated channels with high and low Ca2+ permeability in single rat retinal ganglion cells. AB - The patch-clamp technique was used to record whole-cell currents induced by alpha amino-3-hydroxyl-5-methyl-isoxazol-4-propionic acid (AMPA) or kainate in solitary rat retinal ganglion cells (n = 125) in vitro. Two groups of retinal ganglion cells could be distinguished according to their responses to kainate or AMPA in extracellular solutions with Ca2+ as the only permeant cation. The ratio of the steady-state currents evoked by a given concentration of AMPA compared to kainate was low (0.08) in the first group and high (0.61) in the second group of retinal ganglion cells. The Ca2+ permeability through AMPA/kainate receptor-operated channels was low (PCa2+/PCs+ < 0.1) in the first group (n = 74, 59%) and moderate (PCa2+/PCs+ = 0.53) in the second group (n = 51, 41%) of retinal ganglion cells. The fraction of the total current induced by stimulation of non-N-methyl-D aspartate receptors that is flowing through Ca2+ permeable AMPA/kainate channels in single cells with high Ca2+ permeability was estimated by comparing the current-voltage relationship in extracellular solutions with either Ca2+ or Na+ as the sole charge carrier. The contribution of Ca(2+)-permeable channels to the non-N-methyl-D-aspartate receptor induced whole-cell current in single Ca2+ permeable cells (n = 12) ranged from 40 to 70%, correlating with the intermediate level of Ca2+ permeability (PCa2+/PCs+ = 0.22-0.80) measured by an independent method in these cells. Thus, single Ca(2+)-permeable cells appear to express at least two types of AMPA/kainate receptor-operated channels with high or low Ca2+ permeability. Using the polymerase chain reaction, transcripts for the glutamate receptor subunits 1-4, including their "flip" and "flop" versions, were identified in retinal ganglion cells. Together, these findings suggest that among rat retinal ganglion cells there are differences in the pattern of expression of AMPA/kainate receptor-operated channels. Moreover, individual cells co-express multiple heterologous non-N-methyl-D-aspartate receptors with distinct functional properties. The functional diversity of these receptors may play an important role in controlling Ca2+ entry into neurons. We speculate that the low Ca2+ permeability and the preference for kainate in one group of retinal ganglion cells may be due to the predominant expression of non-N-methyl-D-aspartate receptors containing the edited form of the glutamate receptor subunit 2 flop splice variant. PMID- 7477899 TI - Mu-opioid agonist enhancement of prostaglandin-induced hyperalgesia in the rat: a G-protein beta gamma subunit-mediated effect? AB - Guanine nucleotide-binding regulatory protein stimulation of adenylyl cyclase has been shown to be an important second messenger system for many processes, including mechanical hyperalgesia. Recently, interactions between guanine nucleotide-binding regulatory protein subunits and adenylyl cyclase affecting the level of cyclic adenosine 3',5'-monophosphate accumulation have been demonstrated. In this study we evaluated such an interaction by measuring paw withdrawal thresholds to mechanical stimuli in Sprague-Dawley rats in the presence of two direct-acting hyperalgesic agents, prostaglandin E2 and the adenosine A2-agonist, CGS21680. The effects of two agents expected to liberate inhibitory guanine nucleotide-binding regulatory protein subunits were also studied: [D-Ala2,N-Me-Phe4,Gly5-ol]enkephalin (a mu-opioid receptor agonist) and N6-cyclopentyladenosine (an A1-adenosine agonist). Injection of [D-Ala2,N-Me Phe4,Gly5-ol]enkephalin immediately before prostaglandin E2 or CGS21680 significantly attenuated the hyperalgesia subsequently induced by these agents, i.e. the sensitivity to these hyperalgesic agents was decreased. On the other hand, injection of [D-Ala2,N-Me-Phe4,Gly5-ol]enkephalin 5 min after prostaglandin E2 or CGS21680 significantly enhanced the hyperalgesia observed. Injection of the adenosine A1-agonist N6-cyclopentyladenosine immediately before and 5 min after prostaglandin E2 or CGS21680 had a similar effect to [D-Ala2,N-Me-Phe4,Gly5 ol]enkephalin. The decrease in sensitivity to prostaglandin E2- and CGS21680 induced hyperalgesia by preadministration of [D-Ala2,N-Me-Phe4,Gly5-ol]enkephalin or N6-cyclopentyladenosine and the enhancement by postadministration were all reversed by pertussis toxin, an inhibitor of inhibitory guanine nucleotide binding regulatory protein, suggesting the involvement of an inhibitory guanine nucleotide-binding regulatory protein.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7477900 TI - Disproportionate regulation of nuclear- and mitochondrial-encoded cytochrome oxidase subunit proteins by functional activity in neurons. AB - Cytochrome oxidase is the terminal enzyme in the mitochondrial respiratory chain engaged in oxidative metabolism and energy production. In mammals, the holoenzyme is composed of 13 subunits encoded by both nuclear and mitochondrial genomes. The goal of the present study was to compare the effect of afferent impulse blockade on the expression of these two genomes at the subunit protein level. It also aimed to determine the correlation between the level of cytochrome oxidase activity and the relative amount of subunit proteins. Relative enzyme activity was analysed histochemically, and relative amounts of subunits IV (nuclear encoded) and II/III (mitochondrial-derived) proteins were obtained immunohistochemically by anti-subunit IV and anti-subunit II/III antibodies in the lateral geniculate nucleus and the primary visual cortex of adult monkeys. In the normal visual centers, similar staining patterns were found for all three markers. After three and seven days of tetrodotoxin treatment, levels of enzyme activity and subunit proteins declined disproportionately in the deprived laminae of the visual center. Densitometric analysis indicates that changes in enzyme activity and subunit IV proteins were significantly greater than those of subunit II/III proteins (P < 0.01). The finding that nuclear and mitochondrial genomes are disproportionately regulated at subunit protein levels by neuronal activity implies that the two genomes operate under different regulatory mechanisms. Changes in subunit IV paralleled most closely those of cytochrome oxidase activity (coefficient of determination r2 = 0.95). This suggests that nuclear derived subunit IV protein may play a pivotal role in controlling cytochrome oxidase holoenzyme activity. PMID- 7477901 TI - Fos and serotonin immunoreactivity in the raphe nuclei of the cat during carbachol-induced active sleep: a double-labeling study. AB - The microinjection of carbachol into the nucleus pontis oralis produces a state which is polygraphically and behaviorally similar to active sleep (rapid eye movement sleep). In the present study, using double-labeling techniques for serotonin and the protein product of c-fos (Fos), we sought to examine whether immunocytochemically identified serotonergic neurons of the raphe nuclei of the cat were activated, as indicated by their expression of c-fos, during this pharmacologically-induced behavioral state (active sleep-carbachol). Compared with control cats, which were injected with saline, active sleep-carbachol cats exhibited a significantly greater number of c-fos-expressing neurons in the raphe dorsalis, magnus and pallidus. Whereas most of the c-fos-expressing neurons in the raphe dorsalis were small, those in the raphe magnus were medium-sized and in the raphe pallidus they were small and medium-sized. The mean number of serotonergic neurons that expressed c-fos (i.e. double-labeled cells) was similar in control and active sleep-carbachol cats. These data indicate that there is an increased number of non-serotonergic, c-fos-expressing neurons in the raphe dorsalis, magnus and pallidus during the carbachol-induced state.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7477902 TI - Renshaw cells and recurrent inhibition: comparison of responses to cyclic inputs. AB - Spinal recurrent inhibition influences the discharge patterns of motoneurons and spinal interneurons. The precise pattern of this influence depends on the static and dynamic characteristics of this feedback system. It is thus of importance to quantify its characteristics as well as possible. We here compare nonlinear features (hysteresis) in Renshaw cells and recurrent inhibition in response to cyclic stimulation of motor axons. In pentobarbitone-anaesthetized or decerebrate cats, intracellular recordings were obtained from 26 hindlimb muscle nerves skeleto-motoneurons and extracellular recordings from nine Renshaw cells. Various hindlimb muscle nerves (dorsal roots cut) or ventral roots (dorsal roots intact) were prepared for electrical stimulation to elicit recurrent inhibition in motoneurons or discharges in Renshaw cells. Stimulus patterns consisted of repetitive pulse trains whose rates varied cyclically between around 10 pulses/s and several tens of pulses/s, at modulation frequencies between 0.1 and 1.0 Hz, in one of two waveforms: triangular or sinusoidal. Recurrent inhibitory potentials in motoneurons and discharge patterns of Renshaw cells were averaged with respect to triggers (cycle-triggers) marking a fixed phase in the stimulation cycle. In another two experiments, motor axons to hindlimb muscles (soleus and medial gastrocnemius) were stimulated with sinusoidal and distorted temporal patterns to show their effects on force production. Most often the cycle averaged motoneuron membrane potential changed in a temporally asymmetrical way, i.e. it fairly rapidly hyperpolarized early in the stimulus cycle (during increasing rate) and then depolarized more slowly throughout the rest.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7477903 TI - Purification, characterization and developmental expression of a brain-specific chondroitin sulfate proteoglycan, 6B4 proteoglycan/phosphacan. AB - A large brain-specific chondroitin sulfate proteoglycan, identified with monoclonal antibody 6B4 (6B4 proteoglycan/phosphacan), was isolated from rat brain. Soluble proteoglycans in the phosphate-buffered saline extract from 20-day old rat whole brain were fractionated by anion exchange chromatography and CsCl density gradient centrifugation. 6B4 proteoglycan was further purified by gel filtration and additional ion exchange chromatography. The molecular mass of 6B4 proteoglycan shifted from 800 to 300 x 10(3) mol. wt after chondroitinase ABC digestion. The core protein was substituted with chondroitin sulfate chains with an average molecular weight of 21,000, keratan sulfate and HNK-1 carbohydrates. Glycosidase digestion of 6B4 proteoglycan with O-glycanase, N-glycanase, endo beta-galactosidase, or keratanase did not remove the HNK-1 epitopes. The expression of 6B4 proteoglycan was developmentally regulated in the rat cerebral cortex; appearing first at embryonic day 14, peaking at postnatal day 0, and persisting throughout adulthood at a lower level. Immunohistochemical analysis indicated that 6B4 proteoglycan was distributed along the radial glial fibers and on the migrating neurons in the embryonal rar cerebrum. The radial glial fibers were stained intensely all along their length, but the neurons in the cortical plate were not stained in contrast to the moderate staining of the migrating neurons in the intermediate zone and the subplate. From postnatal day 5 to postnatal day 20, 6B4 proteoglycan was present throughout the cortex. After postnatal day 30, staining of the neuropil was weakened, and the expression of 6B4 proteoglycan was restricted around subsets of neurons. The positive neurons were mostly non-pyramidal cells (> 95%) and were relatively concentrated in layers IV and VI of the primary somatosensory cortex. Immunohistochemical analysis of the dissociated cortical neurons indicated that 6B4 proteoglycan was distributed on the cell bodies and neurites. 6B4 proteoglycan strikingly promoted neurite extension of cortical neurons from embryonic day-16 rat embryos when coated on coverslips as a substrate. 6B4 proteoglycan is a brain-specific chondroitin sulfate proteoglycan which carries keratan sulfate and HNK-1 carbohydrates. The spatiotemporal expression profile and effects on the dissociated cerebral neurons suggest that 6B4 proteoglycan plays important roles in the migration and differentiation of neurons in the immature cortex, and also in the maintenance of subsets of neurons in the mature cortex. PMID- 7477904 TI - Intracellular free calcium responses to protons and capsaicin in cultured trigeminal neurons. AB - Acidic solutions and capsaicin are selective chemical stimuli for nociceptive neurons. The effect of these stimuli on intracellular calcium concentration was analysed in cultured trigeminal neurons of newborn rabbits. Rapid reductions in pH (from 7.4 to 5) evoked a transient rise in intracellular calcium concentration of 270% on average over the basal level (162.5 +/- 3.5 nM; n = 174) in 86% of the neurons. Maximal responses were found at pH 5.5. Proton-induced transients were diminished or abolished by 20 mM CaCl2, by zero CaCl2 and by 1 microM Ruthenium Red. In response to 1 microM capsaicin, 40% of the cells that were sensitive to protons also increased their intracellular calcium concentration to 218% of control. Capsaicin-induced intracellular calcium concentration rises were composed of an initial peak followed by a second, slower intracellular calcium concentration elevation. The capsaicin response was completely blocked by 1 microM Ruthenium Red, and disappeared in zero calcium, but was augmented in high extracellular calcium. Intracellular calcium concentration responses to capsaicin were still observed in neurons whose response to protons was desensitized by sustained exposure to low pH (pH 6.5). Cells surviving a 10-24h capsaicin (10 microM) treatment, still displayed responses to pH reductions. These results suggest that intracellular calcium concentration rises induced by moderate reductions in pH0 and capsaicin occur through different mechanisms. PMID- 7477905 TI - Functional reinnervation of the vasculature of the adult cat paw pad by axons originally innervating vessels in hairy skin. AB - Sympathetic vasoconstrictor neurons that had previously innervated blood vessels in hairy skin were made to reinnervate the vasculature of the hairless skin of the paw pad by suturing the central stump of the cut sural nerve to the distal stump of the cut tibial nerve. After allowing sufficient time for the reinnervation, electrical stimulation of the vasoconstrictor pathway in the lumbar sympathetic trunk produced a reduction of the blood flow that was significantly greater than in control animals. There was also a clear sign of a "denervation supersensitivity" of the blood vessels as evidenced by a significantly increased vasoconstriction that followed the systemic application of the alpha 1-adrenoceptor specific agonist phenylephrine. Neurogenic vasodilation evoked by antidromic excitation of small diameter primary afferent neurones was significantly impaired although myelinated and unmyelinated primary afferents had re-grown into the target tissue. Electrical stimulation of the intact tibial nerve (containing sympathetic vasoconstrictor axons and nociceptive primary afferent fibres) in control animals, always produced vasodilatation indicating that the neurogenic vasodilatation can override the sympathetic vasoconstrictor response. By contrast, electrical stimulation of cross-unioned nerves consistently produced a robust vasoconstriction. We conclude that sympathetic vasoconstrictor neurons have a high capacity to functionally reinnervate autonomic effector organs in the adult cat. Despite this functional recovery, the blood vessels exhibited stronger than normal responses to an alpha 1-adrenoceptor agonist. The impaired neurogenic vasodilatation mediated by small diameter afferents may be due to their poor ability to re-establish their efferent vasodilatory function. Alternatively it may be masked by the strong vasoconstriction. PMID- 7477906 TI - Localization of N-cadherin in the normal and regenerating nerve fibers of the chicken peripheral nervous system. AB - The localization of N-cadherin in the normal, and regenerating nerve fibers was investigated by immunocytochemistry in the chicken sciatic nerve. The normal unmyelinated fibers exhibited N-cadherin immunoreactivity on the plasma membranes of axons and Schwann cells where they were in contact with each other, while myelinated fibers displayed no immunoreactivity except at the mesaxon where Schwann cell plasma membranes were attached to each other. In the regenerating nerves, intense immunoreactivity was demonstrated on the surface of plasma membranes of axons and Schwann cells where axon-axon and axon-Schwann cell contacts were made. No immunoreactivity was observed on the plasma membranes where regenerating axons or Schwann cells were in touch with the basal lamina. In addition, it was revealed that some vesicles in the growth cones had distinct N cadherin immunoreactivity at the inner limiting membrane surface. These findings indicate that N-cadherin may be involved in the axon-axon and axon-Schwann cell adhesion in the normal unmyelinated as well as regenerating nerve fibers, and also in the attachment of Schwann cell processes at the mesaxon of myelinated fibers. In addition, these findings suggest that N-cadherin might be, at least in part, supplied by fusion of growth cone vesicles with the surface plasma membranes in growing axons. PMID- 7477907 TI - Distribution of messenger RNAs for D1 dopamine receptors and DARPP-32 in striatum and cerebral cortex of the cynomolgus monkey: relationship to D1 dopamine receptors. AB - Messenger RNAs for the D1 dopamine receptor and dopamine- and cyclic AMP regulated phosphoprotein of relative mass 32,000 (DARPP-32) were examined by in situ hybridization in the cynomolgus monkey brain. The messenger RNA distribution was compared to the distribution of D1 dopamine receptors using [3H]SCH 23390 autoradiography. In the caudate nucleus and putamen, D1 dopamine receptor messenger RNA-positive cells were unevenly distributed. Clusters of cells with an approximately three-fold higher intensity of labeling, as compared to surrounding regions, were found. Some of these D1 dopamine receptor messenger RNA intensive cell clusters in the caudate nucleus appeared to some extent to be matched to regions of higher intensity of [3H]SCH 23390 binding. The distribution of cells expressing DARPP-32 messenger RNA in the caudate nucleus and putamen was found to be non-clustered. In neocortical regions, cells of different sizes expressing D1 dopamine receptor messenger RNA were present in layers II-VI. D1 dopamine receptor messenger RNA-positive cells were most abundant in layer V. Unexpectedly, no DARPP-32 messenger RNA signal was detected in neocortex. Chronic SCH 23390 administration did not change the relative levels of messenger RNAs for the D1 dopamine receptor and DARPP-32 or [3H]SCH 23390 binding as measured by quantitative image analysis. The clustered distribution of D1 dopamine receptor messenger RNA is in contrast to that of DARPP-32 messenger RNA. This suggests that D1 dopamine receptors may play a more significant role in regulating DARPP 32 function in patch regions as compared to matrix regions. D1 dopamine receptor messenger RNA-expressing cells could also be visualized in several layers of the primate neocortex, implying that dopamine acts through D1 dopamine receptors within functionally different neuronal circuits of the neocortex. PMID- 7477908 TI - Adenosine A2 receptors stimulate c-fos expression in striatal neurons of 6 hydroxydopamine-lesioned rats. AB - The induction of the early-gene c-fos after administration of the adenosine A2a receptor agonist CGS 21680, was studied in the striatum of normal rats or in rats with a unilateral 6-hydroxydopamine lesion of the dopaminergic nigrostriatal neurons. CGS 21680 (2.25 mg/kg) induces c-fos expression in the 6-hydroxydopamine lesioned striatum, while up to 40 mg/kg fails to induce c-fos in the intact striatum or in the striatum of normal rats. Blockade of muscarine receptors by scopolamine (5 mg/kg) partially prevents, and stimulation of dopamine D2 receptors by quinpirole (0.5 mg/kg) completely reverses, CGS 21680-induced c-fos expression in the 6-hydroxydopamine-lesioned striatum. In turn, CGS 21680 partially reverses c-fos expression induced by quinpirole in the lesioned globus pallidus. CGS 21680, in addition, dose-dependently reduces the turning behavior induced by quinpirole (0.5 mg/kg) in 6-hydroxydopamine-lesioned rats. The results suggest that CGS 21680 induces c-fos expression in the striatum through direct and indirect mechanisms related to the ability of A2a receptors to stimulate cyclic AMP formation or acetylcholine release which in turn would activate c-fos through muscarinic receptors. PMID- 7477909 TI - Conditioned dopamine release: dependence upon N-methyl-D-aspartate receptors. AB - The study have investigated the effect of a conditioned emotional response using a contextual cue on dopamine release in the rat nucleus accumbens, measured with in vivo microdialysis, and its inhibition by N-methyl-D-aspartate antagonist dizocilpine maleate. The extracellular level of dopamine in the medial nucleus accumbens markedly increased for up to 40 min when rats were given mild footshock in the testing box. When the rats were returned to the testing box, but not given footshock (conditioned emotional response), there was an immediate and long lasting (80 min) increase in extracellular dopamine. Dizocilpine maleate (50 mumol/l) administered into the nucleus accumbens through the dialysis probe had no significant effect on the immediate increase in dopamine induced by conditioned emotional response but completely prevented the later phase. Dizocilpine maleate had no effect on basal dopamine release in control rats but decreased basal dopamine in rats exposed to footshock 2 h previously. The alpha amino-3-hydroxy-5-methyl-4-isoxazoleproprionate/kainate antagonist 6-cyano-7 nitroquinnoxaline-2,3-dione (100 mumol/l) had no effect on the increase in dopamine release in response to conditioned emotional response. The results indicate that the acquisition of conditioned emotional response causes long lasting changes in the mechanisms involved in the glutamatergic control of dopamine release in the nucleus accumbens. Furthermore glutamate inputs into the nucleus accumbens may also regulate the delayed phase of conditioned dopamine release during expression of conditioned emotional response to a contextural cue through activation of N-methyl-D-aspartate receptors. PMID- 7477910 TI - Volumes of hippocampus, amygdala and frontal lobe in Alzheimer patients with different apolipoprotein E genotypes. AB - An increased frequency of apolipoprotein E E4 allele has been reported in patients with late onset Alzheimer's disease. Apolipoprotein E participates in the transport of cholesterol and other lipids and interferes with the growth and regeneration of both peripheral and central nervous system tissues during development and after injury. Apolipoprotein E is also implicated in synaptogenesis. Apolipoprotein E isoforms differ in binding to amyloid-beta protein and tau protein in vitro. Here, we wanted to study the effect of apolipoprotein E genotype on the magnitude of damage in the hippocampus, where a marked synapse loss exists in Alzheimer's disease. We measured by magnetic resonance imaging the volumes of the hippocampus, amygdala, and frontal lobes in the three Alzheimer subgroups: patients with 2, 1 or 0 E4 alleles. We also investigated the profile of deficits on tests assessing memory, language, visuospatial, executive, and praxic functions of these Alzheimer subgroups. All Alzheimer patients were at early stage of the disease. We found that Alzheimer patients with E4/4 genotype (N = 5) had smaller volumes of the hippocampus and the amygdala than those with E3/4 (N = 9) and those with E3/3 or E2/3 (N = 12). The difference was significant for the right hippocampus (-54% of control) and the right amygdala (-37% of control). The volumes of the frontal lobes were similar across the Alzheimer subgroups. The patients with E4/4 also showed lowest scores on delayed memory tests and differed from E3/3, 3/2 patients in the list learning test (< 0.05).(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7477911 TI - Synchronous development of pyramidal neuron dendritic spines and parvalbumin immunoreactive chandelier neuron axon terminals in layer III of monkey prefrontal cortex. AB - Postnatal development of the primate cerebral cortex involves an initial proliferation and the subsequent attrition of cortical synapses. Although these maturational changes in synaptic density have been observed across the cortical mantle, little is known about the precise time course of developmental refinements in synaptic inputs to specific populations of cortical neurons. We examined the postnatal development of two markers of excitatory and inhibitory inputs to a subpopulation of layer III pyramidal neurons in area 9 and 46 of rhesus monkey prefrontal cortex. These neurons are of particular interest because they play a major role in the flow of information both within and between cortical regions. Quantitative reconstructions of Golgi-impregnated mid-layer III pyramidal neurons revealed substantial developmental changes in the relative density of dendritic spines, the major site of excitatory inputs to these neurons. Relative spine density on both the apical and basilar dendritic trees increased by 50% during the first two postnatal months, remained at a plateau through 1.5 years of age, and then decreased over the peripubertal age range until stable adult levels were achieved. As a measure of the postnatal changes in inhibitory input to the axon initial segment of these pyramidal neurons, we determined the density of parvalbumin-immunoreactive axon terminals belonging to the chandelier class of local circuit neurons. The density of these distinctive axon terminals (cartridges) exhibited a temporal pattern of change that exactly paralleled the changes in dendritic spine density. These results suggest that subpopulations of cortical neurons may be regulated by dynamic interactions between excitatory and inhibitory inputs during development and, in concert with other data, they emphasize the cellular specificity of postnatal refinements in cortical circuitry. PMID- 7477912 TI - Simultaneous expression of excitatory postsynaptic potential/spike potentiation and excitatory postsynaptic potential/spike depression in the hippocampus. AB - Tetanic stimulation of afferents in the stratum radiatum of the CA1 area of the rat hippocampus results in long-term potentiation of excitatory synaptic responses in pyramidal cells. Previous studies have reported a greater increase in the population spike amplitude following the induction of long-term potentiation than could be accounted for by the increase of the slope of the population excitatory postsynaptic potential. Two hypotheses have been proposed to explain this phenomenon (called excitatory postsynaptic potential/spike potentiation): a modification of the firing threshold and/or a modification of the inhibitory drive. Previous studies have not, however, addressed the question of possible changes in spike threshold in association with long-term depression. This paper examines whether the concomitant long-term potentiation of pharmacologically isolated N-methyl-D-aspartate receptor-mediated excitatory postsynaptic potentials, reported previously, is also associated with a change in spike threshold. When the amplitude of the population spike is plotted as a function of the slope of the population excitatory postsynaptic potential (excitatory postsynaptic potential/spike curve), excitatory postsynaptic potential/spike potentiation (depression) is seen as a shift of the excitatory postsynaptic potential/spike curve to the left (right) following a conditioning stimulus. In this study, using kainic acid lesioned hippocampus, we have shown that tetanic stimulation produced excitatory postsynaptic potential/spike potentiation of the control synaptic response and excitatory postsynaptic potential/spike depression of the isolated N-methyl-D-aspartate receptor-mediated responses. PMID- 7477913 TI - Developmental expression of alpha 7 neuronal nicotinic receptor messenger RNA in rat sensory cortex and thalamus. AB - The distribution of alpha 7 messenger RNA expression was characterized in developing rat cortex and thalamus. Northern blot analysis of neonatal and adult cortex revealed a single messenger RNA transcript of 5.7 kb. Using in situ hybridization with both full length and short 35S-labeled alpha 7 riboprobes, a distinct transient expression of messenger RNA within sensory cortex and thalamus, during early postnatal development, was observed. alpha 7 transcripts were expressed in low levels as early as embryonic day 13 in the ventricular zone of the neocortex, and as early as embryonic day 15 in the thalamic neuroepithelium. A marked increase in messenger RNA levels was observed during the late prenatal period in both sensory and non-sensory regions of the cortex and thalamus. Moderate to high levels of messenger RNA were maintained into the first postnatal week, followed by a decline into adulthood. alpha 7 messenger RNA expression was significantly higher in the anterodorsal, lateral dorsal, ventral posterior medial and ventral posterior lateral thalamic nuclei of postnatal day 7 pups than in adult brains. Expression of messenger RNA within dorsal lateral geniculate, ventral lateral geniculate and medial geniculate did not show a significant reduction with age. Within the developing cortex, messenger RNA expression delineated the primary somatosensory, auditory and visual cortices in a unique laminar pattern that was consistently and significantly higher than in the adult in superficial layer VI. Higher levels of expression were also observed in retrosplenial cortex at postnatal day 7 than in the adult. Tangential sections through postnatal day 7 cortex revealed low levels of alpha 7 messenger RNA expression delineating the primary sensory areas in layer IV, corresponding to acetylcholinesterase-labeled thalamocortical afferents. However, these sensory areas exhibited higher levels of alpha 7 messenger RNA expression and were more clearly defined in layer VI, but not by acetylcholinesterase staining. The distribution of alpha 7 messenger RNA within the developing thalamocortical system parallels the distribution of alpha-bungarotoxin binding sites and suggests that the receptor is localized on both thalamic cells and their cortical target neurons. This transient and distinct pattern of distribution of the alpha 7 neuronal nicotinic receptor, which coincides with the major phase of thalamocortical development, suggests that it may play a functional role in the development of cortical circuitry. PMID- 7477914 TI - Altered tonic L-3,4-dihydroxyphenylalanine systems in the nucleus tractus solitarii and the rostral ventrolateral medulla of spontaneously hypertensive rats. AB - We have proposed that L-3,4-dihydroxyphenylalanine (L-DOPA) is a neurotransmitter in the central nervous system [Y. Misu et al. (1995) Adv. Pharmac. 32, 427-459]. L-DOPA as a probable neurotransmitter for the primary baroreceptor afferents tonically functions to mediate cardiodepressor control in the nucleus tractus solitarii and also tonically functions to mediate cardiopressor control in the rostral ventrolateral medulla of rats. We further attempted to clarify whether a transmitter-like L-DOPA system is altered in these areas of adult spontaneously hypertensive rats. By microdialysis in the left nucleus tractus solitarii area, the basal L-DOPA release was lower in spontaneously hypertensive rats than that in Wistar-Kyoto rats. This release was partially reduced by tetrodotoxin (1 microM) to the same absolute levels in the two strains. Tonic neuronal L-DOPA release is impaired in this nucleus of spontaneously hypertensive rats. This impairment is not secondarily due to decrease in formation or increase in decarboxylation of L-DOPA, since tyrosine hydroxylase activity was increased in spontaneously hypertensive rats, compared to Wistar-Kyoto rats, while no difference of L-aromatic amino acid decarboxylase activity was seen in the caudal dorsomedial medulla including the nucleus. L-DOPA (10-300 ng) microinjected into the nucleus produced dose-dependent hypotension and bradycardia. A maximum depressor response of spontaneously hypertensive rats to L-DOPA at higher doses was slightly greater than that of Wistar-Kyoto rats. On the other hand, in the left rostral ventrolateral medulla, the basal L-DOPA release was higher in spontaneously hypertensive rats than that in Wistar-Kyoto rats. This release was also partially reduced by tetrodotoxin to the same absolute levels in the two strains. Tonic neuronal L-DOPA release is enhanced in spontaneously hypertensive rats. This enhancement seems to include partially a decrease in decarboxylation of L-DOPA, since L-aromatic amino acid decarboxylase activity was decreased in spontaneously hypertensive rats compared to Wistar-Kyoto rats, while no difference in tyrosine hydroxylase activity was seen. L-DOPA (10-600 ng) produced dose-dependent hypertension and tachycardia. Importantly, a pressor response of spontaneously hypertensive rats to L-DOPA at lower doses was slightly greater than that of Wistar-Kyoto rats. L-DOPA seems to play a transmitter-like role in blood pressure regulation at levels of the nucleus tractus solitarii and rostral ventrolateral medulla in rats.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 7477915 TI - Axonal regeneration of identified septohippocampal projection neurons in vitro. AB - Previous studies have shown that the septohippocampal projection develops in vitro. It remained unclear, however, whether this in vitro projection resulted from axonal regeneration of septal neurons transected by the culture preparation, or from a priori outgrowth of undamaged cells that had not yet sent an axonal process to the hippocampus at the time of tissue preparation. Here we have prepared slice cultures of identified septohippocampal projection neurons that were retrogradely labeled by hippocampal tracer injection prior to slice preparation. We demonstrate that these identified projection neurons survive axotomy caused by culture preparation, are able to regrow an axonal process in vitro, and innervate a co-cultured hippocampal slice. PMID- 7477916 TI - Activation of dopamine D4 receptor inhibits an L-type calcium current in cerebellar granule cells. AB - The functions of the D4 receptor, a newly cloned D2-like receptor, as well as the identity of cells expressing it, are still poorly defined. Using quantitative polymerase chain reaction we detected the messenger RNA of the D4, but not other D2-like receptor, in cultured granule cells from neonatal rat cerebellum. In these neurons, dopamine reduced high-voltage-activated calcium current, with a pharmacology corresponding to that of the D4 receptor. The response declined from one to three days, when calcium currents were mostly sensitive to nifedipine, to 15 days, when nifedipine-insensitive calcium currents were also present and D4 receptor messenger RNA had declined. The dopamine response was abolished after pretreatment of the cells by pertussis toxin, was potentiated and made irreversible by infusion of guanosine 5'-O-(3-thiotriphosphate) but persisted in the presence of cyclic AMP and isobutylmethylxanthine. These results indicate the presence in the neonatal cerebellum of a functional D4 receptor inhibiting an L type calcium current, an action involving a Gi/Go protein but independent from adenylate cyclase inhibition. PMID- 7477917 TI - Developmental change from inhibition to facilitation in the presynaptic control of glutamate exocytosis by metabotropic glutamate receptors. AB - We have addressed the role of presynaptic metabotropic glutamate receptors in the control of glutamate release from cerebrocortical nerve terminals. The metabotropic glutamate receptor agonist (1S,3R)-1-aminocyclopentane-1,3 dicarboxylic acid enhances the release evoked by a submaximal depolarization in the presence of low concentrations of arachidonic acid and in a staurosporine sensitive manner. In contrast, (1S,3R)-1-aminocyclopentane-1,3-dicarboxylic acid and L(+)-2-amino-4-phosphonobutyrate inhibit the release evoked by a maximal depolarization, in the absence of arachidonic acid and by a staurosporine insensitive mechanism. Interestingly, the effects of the metabotropic glutamate receptors that inhibit glutamate release are only observed in the nerve terminals from young rats (one to three weeks), while the facilitatory effects are better seen in latter developmental stages (three to four weeks) and adult (two to three months) rats, coinciding with the development of the maximal capacity of glutamate uptake. These results indicate the existence of important developmental changes in the presynaptic control of glutamate release. PMID- 7477918 TI - A syntaxin-related protein controls acetylcholine release by different mechanisms in Aplysia. AB - Polyclonal antibodies raised against rat syntaxin-1B and an affinity-purified fraction have been used to study the functional role of this protein in transmitter release from Aplysia neurons. In a ganglionic protein extract, this fraction recognized a 37,000 molecular weight protein which therefore might be the Aplysia homologue of rat brain syntaxin-1B. Immunoglobulins were injected in the presynaptic cell of an identified cholinergic synapse of the buccal ganglion of Aplysia californica. This treatment decreased the postsynaptic response due to a reduction of the number of quanta released in relation to a decline of presynaptic Ca2+ current. When antibodies were applied extracellularly, transmitter release also decreased. In contrast to intracellular injection, this reduction was not accompanied by a decrease of the Ca2+ current but by an increase of presynaptic outward current. When injected in the presynaptic neuron, syntaxin complementary RNA also depressed Ca2+ current and transmission. This work provides evidence that Aplysia neurons express a syntaxin-like protein which is involved in the control of the presynaptic Ca2+ influx triggering acetylcholine release from terminals. This protein appears to have an extracellular segment which might interact with outward current. PMID- 7477920 TI - Interleukin-1 beta induces long-term increase of axonally transported opiate receptors and substance P. AB - Interleukin-1 is known to exert pleiotropic effects in host defence mechanisms and in inflammation. Chronic pain, inflammation and interleukin-1 beta enhance the production of substance P. Recently, axonal transport of opiate receptors was found to increase in rat sciatic nerves in the model of Freund's adjuvant-induced arthritis. Here we show that a single intraplantar injection of interleukin-1 beta is able to enhance the axonal transport of mu and kappa opiate receptors and substance P. Indeed, their accumulation was markedly increased in the proximal part of ligated sciatic nerves, but only in the paw injected with interleukin-1. The time course revealed a delayed onset and, more importantly, a long-term increase lasting at least six days, which is in contrast with the short-term pyrogenic effect of interleukin-1. Pretreatment of rats with capsaicin or administration of dexamethasone completely prevented the interleukin-1 beta effect. The present results suggest that interleukin-1 beta may serve as a mediator to sensitize nociceptors in chronic inflammation and possibly in hyperalgesia through long-term changes in neuronal plasticity. PMID- 7477919 TI - Abnormal perikaryal accumulation of neurofilament light protein in the brain of mice transgenic for the human protein: sequence of postnatal development. AB - Adult mice transgenic for the human form of neurofilament light protein display abnormal perikaryal immunoreactivity for this protein in many regions of the CNS and notably the thalamus. To determine the sequence of development of these anomalies, we have compared normal and transgenic mice of different postnatal ages (P0-P70), using immunocytochemistry with primary antibodies recognizing both murine and human sequence of neurofilament light protein (NR-4) or the human form only (DP5-1-12). In normal mouse brainstem, several nuclei displayed immunoreactive perikarya at P0. The number of these perikarya culminated at P10, followed by a general decrease, some nuclei having lost all perikaryal immunostaining in adults. In transgenic mouse brainstem, the distribution of perikaryal immunoreactivity already resembled at P0 that of P10 in normal mouse, and remained unchanged in adults. Differences between normal and transgenic mice were even more pronounced in the forebrain. Some nuclei of normal mouse basal forebrain that were weakly immunopositive at P10 or P20, but no longer in adults, were already labeled at P0 and remained so or became more intense at later stages in transgenic mice. In the thalamus of normal mouse, perikaryal labeling was faint, confined to a few nuclei, and detected only transiently at P10, whereas in transgenics, it was already observed in some nuclei at P0, increased in intensity and extended to other nuclei at P10, and persisted thereafter. Strongly immunoreactive, inflated perikarya with excentric nuclei were prominent in these thalamic nuclei at P20, and even larger in size at P70. In the cerebral cortex of normal mice, layers II-III and layer V of many cytoarchitectonic areas showed immunoreactive cell bodies at P10, a distribution which became gradually restricted to the parietal cortex in adults. In transgenic mice, immunopositive cortical cell bodies were first detected at P3, filled layers II-III of numerous cortical areas at P10, and then rapidly decreased in number to approach the adult pattern at P20. In the cortex as well as thalamus of P10 transgenic mice, differences between the patterns of cellular staining with clones NR4 and DP5-1 12 antibodies indicated that both the murine and human proteins were accumulated in these neurons. Thus, neurofilament light protein accumulation in the transgenic mouse brain generally involved neurons displaying perikaryal immunoreactivity for the protein at least at some point during normal postnatal development.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 7477921 TI - N-methyl-D-aspartate R1 messenger RNA and [125I]MK-801 binding decrease in rat spinal cord after unilateral hind paw inflammation. AB - Recent evidence suggests that N-methyl-D-aspartate receptors play an important role in the etiology and maintenance of chronic nociception. Previous studies have demonstrated that tissue injury or stimulation of nociceptive afferent projections results in the expansion of receptive fields, hyperalgesia and C fiber-induced wind-up, events that can be inhibited by N-methyl-D-aspartate antagonists. This study examines the effect of unilateral hind paw inflammation on N-methyl-D-aspartate R1 messenger RNA and [125I]dizocilpine maleate binding in the L4-L5 segments of the lumbar spinal cord of rats. Spinal cords were examined at 7.5 h, three, seven and 20 days after injection of the left hind paw with 120 microliters of complete Freund's adjuvant. N-methyl-D-aspartate R1 messenger RNA, as measured with in situ hybridization, was observed to decrease bilaterally in laminae I, II and X of the lumbar spinal cord. This decrease was evident in laminae I and II at 7.5 h and three days after hind paw injection. In lamina X, a postinjection decrease in hybridization signal was observed at 7.5 h and seven days. A bilateral decrease in [125I]dizocilpine maleate binding was observed in laminae I and II at three, seven and 20 days after paw injection. This observed decrease in binding at the N-methyl-D-aspartate receptor suggests a compensatory mechanism by which N-methyl-D-aspartate-mediated nociceptive events may be modulated. PMID- 7477922 TI - Complement and clusterin in the spinal cord dorsal horn and gracile nucleus following sciatic nerve injury in the adult rat. AB - We provide evidence for activation of the complement cascade in the dorsal horn of the spinal cord and in the gracile nucleus in the brainstem following sciatic nerve transection in the adult rat. Immunocytochemical analyses showed immunoreactivity for endogenous immunoglobulin G as shown by immunostaining with F(ab')2 antibodies, as well as complement factors C1, C1q, C3, C3d and C9 in the appropriate central termination areas of the injured sciatic nerve. Results from double labelling immunocytochemistry showed a strong association between immunoglobulin and complement factors on the one hand and reactive microglia on the other. However, some complement immunoreactivity was also found in the neuropil, possibly representing secreted complement. In situ hybridization with an oligonucleotide probe showed a marked increase in C3 messenger RNA, indicating local synthesis of C3 protein. In parallel with activation of complement, there was an increased immunoreactivity for the putative complement inhibitor clusterin, which co-localized with glial fibrillary acidic protein-positive astrocytes. In situ hybridization showed an increased labelling of clusterin messenger RNA. These findings indicate that complement activation and up regulation of complement inhibitors are prominent central responses to peripheral sensory nerve injury. These responses may therefore be important elements underlying so-called transganglionic degenerative changes in primary sensory axons and terminals. PMID- 7477923 TI - An immunohistochemical study of Ca2+/calmodulin-dependent protein kinase IV in the rat central nervous system: light and electron microscopic observations. AB - We observed the distribution pattern of Ca2+/calmodulin-dependent protein kinase IV in rat brain and spinal cord using an immunohistochemical method by light and electron microscopy. Particularly strong immunoreactivity was detected in the telencephalic structures such as the olfactory bulb, cerebral cortex, hippocampal formation, caudate-putamen, most nuclei of the dorsal thalamus and the granule cell layer of the cerebellum. Relatively weak staining was observed in the amygdaloid body, some neuron groups of the brainstem reticular formation, the inferior olivary nucleus and the posterior horn of the spinal cord. Immunohistochemical reactivity was not detected in the globus pallidus, substantia nigra, sensory and motor nuclei of the cranial nerves, or in the spinal cord anterior horn. Overall, the distribution of Ca2+/calmodulin-dependent protein kinase IV-like immunoreactivity broadly paralleled the sites of expression of signals for messenger RNA of this enzyme. At the subcellular level, Ca2+/calmodulin-dependent protein kinase IV-like immunoreactivity appeared exclusively in the nuclei of neurons in the various brain regions, and immunopositive reactivity, although less strong, was also observed in dendritic processes, as well as on the granular endoplasmic reticulum in neuronal somata in these areas. Axon terminals, however, did not show immunoreactivity. These studies demonstrate that Ca2+/calmodulin-dependent protein kinase IV-like immunoreactivity is distributed widely in the central nervous system. The significance of the localization of this enzyme in nuclei is discussed in relation to gene expression. PMID- 7477925 TI - Pyramidal tract and corticospinal neurons with branching axons to the dorsal column nuclei of the cat. AB - Extracellular single activity was recorded from pericruciate neurons in anaesthetized, paralysed, artificially ventilated cats. A total of 309 neurons were identified antidromically by stimulation of the dorsal column nuclei (229 from the nuneate nucleus and 80 from the gracile nucleus). The study addressed the question whether pericruciate-dorsal column nuclei neurons (corticonuclear cells) sent collaterals to the ipsilateral red nucleus and/or to the contralateral nucleus reticularis gigantocellularis. Also, the ipsilateral pyramidal tract was stimulated at mid-olivary level, as was the crossed corticospinal tract at C2, Th2 and L2 levels in order to know whether the corticonuclear cells sent their axons to the spinal cord and if so to which level. It was found that more than 95% of the corticonuclear fibres coursed through the pyramidal tract. A significant (28.4%; 88/309) proportion of the the corticonuclear neurons sent collaterals to the red nucleus and/or to the nucleus reticularis gigantocellularis. About 68% (209/309) of the corticonuclear cells did not send their axons to the spinal cord and the remainder were corticospinal neurons. Most of the corticospinal fibres terminated at the cervical level (72/100) and the remaining ended at thoracic (18/100) and lumbar (10/100) segments of the cord. While 63.4% (123/194) of the corticonuclear fibres coursing through the pyramidal tract and ending at supraspinal levels were slow conducting, the great majority of the corticospinal neurons were fast conducting (91/100). The non-corticospinal neurons were significantly slower conducting than the corticospinal cells. The corticogracile neurons were slower conducting than the corticocuneate cells. Of the 88 corticonuclear neurons that sent at least a branch to the sites tested, 50% branched into the red nucleus, 35.2% into the nucleus reticularis gigantocellularis and 14.7% into both nuclei, without significant difference between non-corticospinal and corticospinal cells. Most of the main axons of the corticonuclear cells ended at bulbar and cervical levels (281/309 or 90.9%). The data indicate that pericruciate-dorsal column nuclei neurons form a particular substrate within pyramidal tract cells. They can serve precise functions in motor coordination associated with the selection of their own sensory input. The results are discussed from this point of view. PMID- 7477924 TI - Synaptic loss in cognitively impaired aged rats is ameliorated by chronic human nerve growth factor infusion. AB - In the present study, we assessed the synaptic changes in aged impaired and unimpaired rats, and the effect of exogenous human nerve growth factor administration on behavioral activity and synaptic density. Human nerve growth factor was administered into the rat ventricles with a cannula connected to an osmotic pump in adult, aged impaired and unimpaired rats. Behavioral performance was evaluated in the Morris water maze. Aged impaired rats had an 18 +/- 4% decrease in the number of synaptophysinimmunoreactive presynaptic terminals as compared to aged unimpaired rats. After a continuous four-week human nerve growth factor, the aged impaired rats displayed a significant 16 +/- 3% increase in the number of synaptophysin-immunoreactive presynaptic terminals in the frontal cortex, as compared to aged impaired rats treated with vehicle. This increase correlated with an improvement in water maze performance (r = -0.74, P < 0.001). Measurements of synaptophysin-immunoreactive presynaptic terminals in other cortical and subcortical regions did not show any statistically significant difference or correlations among the various groups. These results support the possibility that nerve growth factor mediates the induction of other trophic factors which, in turn, might potentially produce a sprouting response of non cholinergic fibers that ameliorate the cognitive deficits in impaired, aged rats. PMID- 7477926 TI - GABAergic axons in the ventral forebrain of the rat: an electron microscopic study. AB - The ventral forebrain, including the ventral striatum, the ventral pallidum and the substantia innominata, is an important region involved in the functions of the basal ganglia and the limbic system, as well as the magnocellular corticopetal neurons of the nucleus basalis of Meynert. Although previous studies have shown that this region is richly innervated by GABAergic fibers, little is known with respect to the relative densities of GABAergic to non-GABAergic axon terminals in this region. To address this issue, we have developed a specific rabbit antiserum to GABA and used a postembedding immunocytochemical reaction to analyse the distribution of GABA-like immunoreactive axon terminals in the rat ventral striatum, ventral pallidum and substantia innominata. Of all axon terminals that form identifiable synapses within single ultrathin sections taken from these regions, 11.6% in the ventral striatum, 85.5% in the ventral pallidum and 64.8% in the substantia innominata were GABAergic. Differences were also found in the distribution patterns of these terminals with respect to the size of their synaptic target dendrites. These findings are consistent with previous findings that a majority of inputs to the ventral striatum are excitatory, and that a majority of inputs to the ventral pallidum are inhibitory. Our results provide a first approximation of the anatomical substrate for the physiology and pharmacology of GABA actions in the ventral forebrain region. These results also show that GABA may play an important role in the substantia innominata, where both the cholinergic and the non-cholinergic magnocellular corticopetal neurons reside within a neuropil innervated by many different non-cholinergic fibers. PMID- 7477927 TI - Effect of centrally administered neuropeptide Y on hypothalamic and hypophyseal proopiomelanocortin-derived peptides in the rat. AB - In a previous study, we have shown that neuropeptide Y inhibits the release of alpha-melanocyte-stimulating hormone from the rat hypothalamus in vitro. The aim of the present study was to investigate the possible effect of neuropeptide Y on the regulation of proopiomelanocortin-derived peptides in vivo. Rats received acute or chronic administration of neuropeptide Y in the lateral ventricle and the amount of alpha-melanocyte-stimulating hormone was measured in the hypothalamus and in the neurointermediate lobe of the pituitary. In the same experiments, the amounts of corticotropin-releasing factor and corticotropin were quantified in the hypothalamus and anterior pituitary, respectively. Acute treatment with synthetic neuropeptide Y (0.1 to 10 micrograms/rat) did not modify the amount of alpha-melanocyte-stimulating hormone in the hypothalamus. In contrast, chronic infusion of neuropeptide Y (1.25 micrograms/h) over a seven day period significantly decreased the hypothalamic content of alpha-melanocyte stimulating hormone, suggesting that neuropeptide Y regulates the synthesis and/or the processing of proopiomelanocortin. Concurrently, we found that both acute and chronic infusion of neuropeptide Y induced a significant reduction in corticotropin-releasing factor in the hypothalamus as well as a significant decrease in alpha-melanocyte-stimulating hormone and corticotropin in the neurointermediate and anterior lobes, respectively. Quantitative in situ hybridization histochemistry showed that chronic administration of neuropeptide Y also caused a reduction of proopiomelanocortin messenger RNA levels both in the intermediate and anterior lobes of the pituitary. Administration of neuropeptide Y (10(-6) M) on perifused rat hypothalamic slices caused a significant increase in corticotropin-releasing factor release.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7477928 TI - Kinesthetic reference for human orthograde posture. AB - Humans with occluded vision were subjected to superslow tilts of the supporting platform, producing the inclination of the subject's body in the sagittal plane, but subthreshold for the most vestibular and proprioceptive phasic reactions. Two types of perturbation were used: sinusoidal tilts (frequency 0.007 Hz, amplitude 1.5 degrees) and ramps (amplitude 1.0 and 0.25 degrees, angular velocity 0.04 degrees/s). During slow sinusoidal tilts of the platform, the ankle angle and body position undergo periodical changes, but these changes have significant phase lead relative to the platform movement: 119 +/- 26 for ankle angle and 55 +/- 19 degrees for body sway. Gains were about 0.9 for both parameters. Large phase shift (tens of seconds) indicated a long delay in compensation of body inclination by ankle joint. The ramp tilt produced an initial body deviation followed by a slow (seconds or tens of seconds) approach of body position to a new steady level after the termination of ramp. Large slow body movements were superimposed with small irregular oscillations (about 10% of the amplitude of large displacements) of higher frequency. These oscillations resembled normal stabilograms on a stationary support. Thus, the usual process of stabilization of body gravity center was continued, though not around a fixed set-point but relative to a slowly changing position. Data obtained support the hypothesis that, besides operative control assigned to compensate deviations from a reference position, the system of postural control includes at least one additional level, which elaborates this reference using information about mutual position of body links, muscular torques and interaction with the support on the basis of criteria taking into account the energy cost of standing and demands for stability and security. PMID- 7477929 TI - Developmentally regulated vitronectin influences cell differentiation, neuron survival and process outgrowth in the developing chicken retina. AB - Vitronectin is a multifunctional protein involved in the regulation of the immune system and blood coagulation. Here we report that the expression of vitronectin is developmentally regulated in the embryonic retina of the chicken. Vitronectin immunoreactivity was detected in chicken retinas from embryonic day 5, encompassing the cell bodies of most neuroepithelial cells. At this developmental stage, alpha v integrin subunit expression was distributed across the retina, suggesting a ligand/receptor interaction. Expression of both vitronectin and alpha v increased during development and reached a maximum at embryonic day 9, a time when most differentiated neurons grow processes and initiate synapse formation. At this age, vitronectin immunoreactivity appeared to be located predominantly in the fiber and inner plexiform layers of the differentiated stratified retina. alpha v immunoreactivity and mRNA expression was seen associated with all layers formed by differentiated neurons, being most abundant in the ganglion cell and inner nuclear layers. Later in development, levels of vitronectin decreased and immunoreactivity appeared exclusively associated with the fiber layer. In accordance with this pattern of expression, vitronectin as a substrate sustained both proliferation and differentiation of cultured neuroepithelial cells from embryonic day 5 retinas. At later stages, vitronectin supported survival and neurite outgrowth of most differentiated neurons. Our data suggest that vitronectin is a ubiquitous component of the retinal extracellular matrix, serving as a substrate for developmental processes such as proliferation, differentiation of neuron progenitors, cell survival, and axonal and dendritic growth of differentiated neurons. PMID- 7477930 TI - Nitric oxide synthase and chemical coding in cat sympathetic postganglionic neurons. AB - Nitric oxide synthase-like immunoreactivity was found in a subpopulation of sympathetic postganglionic neurons in the cat stellate and lower lumbar ganglia. In the ganglia of other segments such cells were rare. Double staining for tyrosine hydroxylase-like immunoreactivity and nitric oxide synthase-like immunoreactivity or the reduced nicotinamide adenine dinucleotide phosphate diaphorase reaction indicated that nitric oxide synthase-like immunoreactivity and reduced nicotinamide adenine dinucleotide phosphate diaphorase reactivity was always co-localized and was confined to tyrosine hydroxylase-negative (presumably cholinergic) ganglion cells, and was present in most of them. The occurrence of nitric oxide synthase in two subpopulations of cholinergic postganglionic neurons was investigated in triple staining experiments. Presumptive sudomotor neurons have been previously defined as scattered cells containing calcitonin gene related peptide-like immunoreactivity, usually accompanied by vasoactive intestinal peptide-like immunoreactivity: 99% of these contained nitric oxide synthase. Presumptive muscle vasodilator neurons have been previously identified as clumped cells with strong vasoactive intestinal peptide-like immunoreactivity but no calcitonin gene-related peptide-like immunoreactivity: 70% of these contained nitric oxide synthase. Sweat glands were found in the paw pad skin surrounded by varicose fibres showing calcitonin gene-related peptide-like immunoreactivity and vasoactive intestinal peptide-like immunoreactivity, confirming previous work. Such fibres also stained for nitric oxide synthase-like immunoreactivity and reduced nicotinamide adenine dinucleotide phosphate diaphorase reactivity, although their staining was relatively weaker than in the corresponding cell bodies. Varicose fibres with the same chemical coding were also found around all large and most medium and small arteries in the paw skin as well as around arteriovenous anastomoses. Fibres with the muscle vasodilator coding (vasoactive intestinal peptide-like immunoreactivity without calcitonin gene-related peptide-like immunoreactivity) were not seen in paw skin. These results suggest that nitric oxide may act as a co-transmitter (with acetylcholine, substance P, vasoactive intestinal peptide and calcitonin gene related peptide) in sudomotor neurons and (with acetylcholine and vasoactive intestinal peptide) in vasodilator neurons. Collateral branches of sudomotor neurons may innervate skin vessels, and release vasodilator transmitters including nitric oxide to cause the vasodilatation which provides the fluid supply for sweat formation. Alternatively, separate vasodilator neurons to skin may share the same chemical code as sudomotor neurons. PMID- 7477931 TI - Catecholamine secretion, calcium levels and calcium influx in response to membrane depolarization in bovine chromaffin cells. AB - In this paper, we show that exposure of chromaffin cells to high K+ (75 mM) for 5 min releases about 15% of total norepinephrine and 8% of total epinephrine contained in chromaffin cells. The measured resting membrane potential of these cells was -55 mV. Long (10 s) depolarizing electrical pulses applied from a holding potential of -55 mV to 5 mV, that would produce a depolarization similar to exposure to high K+ (75 mM), induced an inward Ca2+ current that inactivated with a time constant of about 0.8 s and promoted the influx of about 1 fmol of Ca2+ into the cell. Both high K+ and electrically-induced depolarization increased intracellular Ca2+ levels to a similar value (about 350 nM). Extrapolation would indicate that total Ca2+ influx in high K+ (75 mM)-stimulated 10(6) chromaffin cells would amount to 1 nmol which would promote the secretion of about 4.9 nmol of norepinephrine and 3.5 nmol of epinephrine from 10(6) chromaffin cells. The results indicate that Ca2+ influx in response to depolarization is short-lived, likely due to Ca(2+)-dependent inactivation of voltage-dependent Ca2+ channels. However, intracellular Ca2+ levels remain high as long as depolarization is present and long after Ca2+ influx has ceased. This would suggest that some processes related to either Ca2+ buffering or extrusion from the cell may be voltage dependent. PMID- 7477932 TI - Differential expression of p140trk, p75NGFR and growth-associated phosphoprotein 43 genes in nucleus basalis magnocellularis, thalamus and adjacent cortex following neocortical infarction and nerve growth factor treatment. AB - A loss of target-derived neurotrophic factors is hypothesized to be one of the major determinants of central nervous system neuronal degeneration. In order to obtain further insight into early neuronal responses to injury, lesion-induced alterations in the expression of high- and low-affinity nerve growth factor receptors, as well as growth-associated phosphoprotein-43 genes in nucleus basalis magnocellularis, thalamic and neocortical neurons were studied. For this purpose, unilateral cortical devascularization operations were conducted on adult rats. Animals received i.c.v. infusions of vehicle or nerve growth factor (12 micrograms/day) and were killed at one, three, seven and 15 days post-lesion. In situ hybridization studies using 35S-labelled oligonucleotide probes for p75NGFR, p140trk and growth-associated phosphoprotein-43 messenger RNAs reveals that these genes were differentially regulated following the lesion. In the nucleus basalis magnocellularis ipsilateral to the lesion, p140trk gene expression significantly decreased on days 3 and 7, while p75NGFR messenger RNA initially increased on day 3 and decreased on days 7 and 15 after lesion. GAP-43 messenger RNA levels were significantly increased in the nucleus basalis magnocellularis on post-lesion days 3 and 7. Moreover, in contrast to p75NGFR or 140trk, growth-associated phosphoprotein-43 messenger RNA levels were significantly increased in pyramidal neurons located in the remaining cortex adjacent to the cortical lesion at all time points. In the lateral and ventroposterior nuclei of the thalamus, growth associated phosphoprotein-43 messenger RNA level was slightly increased on days 1 and 3 and was dramatically decreased, significantly below the levels in sham operated controls, on post-lesion days 7 and 15. During nerve growth factor application, the level of p140trk messenger RNA in the lesioned nucleus basalis magnocellularis returned to values observed in the contralateral nucleus basalis magnocellularis while p75NGFR messenger RNA was increased above values noted in all animals not treated with nerve growth factor. Nerve growth factor treatment did not affect the expression of growth-associated phosphoprotein-43 messenger RNA in any of the areas studied. p140trk messenger RNA was not up-regulated during the time that nerve growth factor was applied, as observed for p75NGFR, but only eight days after interrupting nerve growth factor treatment. Three cell types, nucleus basalis magnocellularis, cortical pyramidal and thalamic neurons, were probably affected in different ways by the devascularization with respect to lesion extent. Consequently, the remaining number of synaptic contacts in each of these brain areas is most likely different which may lead to a differential regulation of growth-associated phosphoprotein-43 messenger RNA.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 7477933 TI - Induction of long-lasting potentiation in the secondary somatosensory cortex by thalamic stimulation requires cortico-cortical pathways from the primary somatosensory cortex. AB - We have investigated the role of cortico-cortical inputs from the primary somatosensory cortex in the induction of long-lasting potentiation in the secondary somatosensory cortex. Long-lasting potentiation of evoked potentials in the feline secondary somatosensory cortex is induced by high frequency stimulation of the ventral posterolateral thalamic nucleus. The secondary somatosensory cortex receives two projections from the ventral posterolateral thalamic nucleus; a direct pathway from the ventral posterolateral thalamic nucleus and a cortico-cortical pathway via the primary somatosensory cortex. The present study was designed to examine dominance of these pathways in the induction of long-lasting potentiation in the secondary somatosensory cortex. Long-lasting potentiation was evaluated by changes in the amplitude of field potentials and current source densities elicited by ventral posterolateral thalamic nucleus test stimulation (0.1 Hz) following conditioning stimulation. The conditioning stimulation, consisting of 20 trains of 200 Hz bursts, was delivered to the ventral posterolateral thalamic nucleus or the primary somatosensory cortex. Field potentials in the secondary somatosensory cortex were simultaneously recorded at 16 points placed vertically at 150 microns intervals from the cortical surface and current source density was computed using these field potentials. First, we blocked inputs from the primary somatosensory cortex to the secondary somatosensory cortex by intracortical injection of lidocaine into the primary somatosensory cortex. The amplitudes of the field potentials recorded in the secondary somatosensory cortex diminished within 5 min after lidocaine injection. Current source density analysis showed a marked decrease in the sink currents in layers II/III (at depths of 450-600 microns).(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7477934 TI - Increased expression and subcellular translocation of the mitogen activated protein kinase kinase and mitogen-activated protein kinase in Alzheimer's disease. AB - The sequential activation of the mitogen-activated protein kinase kinase and its substrate, the mitogen-activated protein kinase is involved in a cascade of protein kinases which link a number of cell surface signals to intracellular changes in enzyme activity and gene expression. In vitro, mitogen-activated protein kinase is able to phosphorylate the microtubule-associated protein tau at Ser-Pro and Thr-Pro sites, thereby generating abnormally hyperphosphorylated tau species that are similar to paired helical filament-tau found in Alzheimer's disease. In the present study, we analysed the levels of immunoreactive mitogen activated protein kinase kinase and mitogen-activated protein kinase in the temporal cortex (area 22) of patients with Alzheimer's disease by means of enzyme linked immuno-sorbent assays and compared these changes with the content of abnormally phosphorylated paired helical filament-tau. The levels of immunochemically detected mitogen-activated protein kinase kinase and mitogen activated protein kinase were both increased in Alzheimer's disease by between 35 and 40% compared with age-matched controls. Elevation of mitogen-activated protein kinase kinase was most pronounced during early stages of Alzheimer's disease and was inversely related to the tissue content of abnormally phosphorylated paired helical filament-tau. Pronounced immunoreactivity of mitogen-activated protein kinase kinase and mitogen-activated protein kinase was present in both tangle bearing neurons and unaffected neurons of the temporal cortex. Immunoreactive neurons were most often localized in the direct vicinity of neuritic plaques. In Alzheimer's disease, the subcellular distribution of mitogen-activated protein kinase kinase and mitogen-activated protein kinase showed a striking translocation from the cytoplasmic to the nuclear compartment. It is suggested that the activation of the mitogen-activated protein kinase cascade which appears to be an early feature of Alzheimer's disease might be critically involved in self-stimulating processes of neurodegeneration and aberrant repair under these conditions. PMID- 7477936 TI - An immunohistochemical study of heat shock protein-27 in the hippocampus in a gerbil model of cerebral ischemia and ischemic tolerance. AB - Preconditioning of the gerbil brain with a 2-min period of sublethal ischemia protects against neuronal damage following a subsequent 3-min period of ischemia which normally damages CA1 neurons of the hippocampus (ischemic tolerance). In this study, we investigated the role of a small stress protein, heat shock protein-27, in the induction of ischemic tolerance. For this purpose, we used immunohistochemistry with an antibody against heat shock protein-27. Normal hippocampus contained very low levels of heat shock protein-27. The preconditioning ischemia for 2 min caused little changes in the heat shock protein-27 immunostaining in CA1 neurons but an increase in heat shock protein-27 immunostaining in a small number of astrocytes in the CA3 region and in many astrocytes in the dentate hilus. The second ischemia for 3 min caused no specific changes in heat shock protein-27 immunostaining in CA1 neurons both with and without tolerance in early reperfusion periods. After seven days, destruction of CA1 neurons occurred in animals without preconditioning and reactive astrocytes were intensely immunostained for heat shock protein-27. An intense heat shock protein-27 immunostaining was also seen in astrocytes in the dentate hilus after the second ischemia in both groups. Thus, we observed no temporal correlation between the induction of heat shock protein-27 and the manifestation of ischemic tolerance in the CA1 neurons. Most intense heat shock protein-27 immunostaining was observed in reactive astrocytes that accumulated in the damaged CA1 region and dentate hilus.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7477935 TI - Induction of heparin-binding growth-associated molecule expression in reactive astrocytes following hippocampal neuronal injury. AB - Heparin-binding growth-associated molecule is a potent neurotrophic factor. To obtain a better understanding of its role in the central nervous system, we studied the changes of its expression in adult rat brain after two types of neuronal injury. In the control hippocampus, expression of heparin-binding growth associated molecule messenger RNA was confined to CA1 pyramidal neurons and some hilar cells. Following transient forebrain ischaemia, the messenger RNA expression decreased within the first two days. On day 4, however, both the messenger RNA level and the number of expression-positive cells markedly increased in the CA1 subfield, where the selective neuronal losses were seen following ischaemia. Double-staining with a heparin-binding growth-associated molecule complementary RNA probe and an anti-glial fibrillary acidic protein antibody revealed that most of the expressing cells were reactive astrocytes. Moreover, the protein induction of heparin-binding growth-associated molecule after neuronal injury was demonstrated by immunohistochemistry using the affinity purified antibodies. This molecule was also induced after intraventricular kainate injection, which is known to cause selective pyramidal cell necrosis in the CA3 region. Four days after the insult, the number of cells expressing the messenger RNA prominently increased in the CA3 subfield ipsilateral to the injection. As observed after the ischaemic insult, most of the expression positive cells were identified as astrocytes. The data presented here suggest that heparin-binding growth-associated molecule, produced by the reactive astrocytes, may play important roles in the repair process after neuronal injury. PMID- 7477937 TI - Seizure related changes in the regulation of opioid genes and transcription factors in the dentate gyrus of rat hippocampus. AB - An in situ hybridization study showed that limbic seizures induced by kainate strongly augmented the prodynorphin and proenkephalin messenger RNA levels in granular cells of the rat hippocampal dentate gyrus. Pentylenetetrazole increased the level of proenkephalin messenger RNA, but slightly decreased that of prodynorphin messenger RNA in the dentate gyrus. Administration of kainate to rats caused a profound increase in messenger RNAs of the transcription factor genes c-fos and c-jun in the dentate gyrus, followed by an increase in the level of the transcriptional complex activator protein-1 in hippocampal neurons. Pentylenetetrazole also elevated the formation of activator protein-1, but the effect appeared earlier than that induced by kainate. Thus, recurrent limbic seizures activate both prodynorphin and proenkephalin genes, whereas generalized clonic-tonic seizures seem to decrease the prodynorphin and increase the proenkephalin gene expression in the dentate gyrus. Furthermore, our present results suggest that the transcription factors, c-fos, c-jun and activator protein-1 complex may be involved in the process of inducing the hippocampal proenkephalin gene, while these factors might be differently involved in regulation of prodynorphin gene expression. PMID- 7477938 TI - A single injection of amphetamine or methamphetamine induces dynamic alterations in c-fos, zif/268 and preprodynorphin messenger RNA expression in rat forebrain. AB - In this study, the effects of a single dose of the indirect dopamine agonists amphetamine and methamphetamine on behavior and messenger RNA expression were evaluated. Expression of c-fos, a member of the leucine zipper family, zif/268 (NGFI-A, egr1 and Krox-24), a member of the zinc finger family, and the opioid peptide, preprodynorphin, was investigated in various regions of rat forebrain with quantitative in situ hybridization histochemistry 1, 2, 3, 6 or 30 h after injection. Behavioral observations indicated that a qualitatively different behavioral syndrome was induced following methamphetamine (15 mg/kg, i.p.) as compared with that observed after amphetamine (5 mg/kg, i.p.). Similarly, methamphetamine induced a different pattern of c-fos and zif/268 messenger RNA induction in sensory/motor cortex, dorsal striatum (caudatoputamen) and ventral striatum (nucleus accumbens) than did amphetamine. The increase in c-fos messenger RNA expression peaked at 1 h and returned to basal levels in all regions by 3 h. In contrast, the increase in zif/268 messenger RNA expression in the cortical regions was equally strong at 1 and 2 h, gradually returning to basal levels by 6 h after either drug. However, in the striatal regions, zif/268 messenger RNA levels peaked at 1 h and declined gradually to basal levels by 6 h. Interestingly, methamphetamine caused an actual suppression of zif/268 gene expression (> 50%) in both caudatoputamen and nucleus accumbens at 3 h. Preprodynorphin messenger RNA expression was increased in a patchy motif in the caudatoputamen and nucleus accumbens beginning at 2 h and returning to basal levels by 30 h after injection of either drug. This study, together with our recently published observation that preprodynorphin messenger RNA is induced in the caudate 3, 6 and 18 h after amphetamine or methamphetamine injection, provides a detailed dynamic description of the differential modulation of c-fos, zif/268 and preprodynorphin messenger RNA expression in the cerebral cortex and striatum by amphetamines over time. These data implicate immediate early gene and preprodynorphin gene expression in the differential response of medium spiny striatal neurons to methamphetamine and amphetamine. PMID- 7477939 TI - Induction of c-fos and zif/268 gene expression in rat striatal neurons, following stimulation of D1-like dopamine receptors, involves protein kinase A and protein kinase C. AB - Changes in the level of dopaminergic activity in the rat striatum lead to the induction of a number of immediate-early genes, including c-fos and zif/268. These immediate-early genes are thought in turn to alter the rate of transcription of downstream genes. There is evidence that the dopaminergic activation of the c-fos and zif/268 genes in the striatum in vivo is linked to stimulation of D1-like dopamine receptors. We have used primary cultures of embryonic rat striatal neurons to identify the intracellular pathways involved in this response. Dopamine (10 nM-5 microM) caused a marked increase in the levels of c-fos mRNA and zif/268 mRNA in cultured striatal neurons, an effect that was reproduced by the D1-like dopamine receptor agonist SKF38393 (10 nM-5 microM). These actions were attenuated by the D1-like antagonist SCH23390 (1 microM) but not by the D2-like antagonist eticlopride (1 microM). The D2-like agonist quinpirole did not increase zif/268 mRNA above basal levels at concentrations up to 5 microM, but caused a slight increase in the levels of c-fos mRNA. The stimulation of c-fos mRNA levels caused by 1 microM SKF38393 was reduced by 45% following pretreatment with the selective protein kinase A inhibitor KT5720, and by 87% following pretreatment with the selective protein kinase C inhibitor calphostin C. The stimulation of zif/268 mRNA levels caused by 1 microM SKF38393 was reduced by 90% following pretreatment with KT5720, but was not significantly affected by pretreatment with calphostin C. In addition, the actions of SKF38393 to stimulate the expression of both immediate-early genes were attenuated by coadministration of quinpirole. These results suggest that SKF38393 acts on striatal neurons to stimulate c-fos expression predominantly through protein kinase C, but also partially through protein kinase A. Conversely, SKF38393 induces zif/268 expression through protein kinase A. The ability of quinpirole to antagonize the actions of SKF38393 on cultured neurons is consistent with the presence of both D1-like receptors on the same neuronal population. PMID- 7477940 TI - Rationale for intrastriatal grafting of striatal neuroblasts in patients with Huntington's disease. AB - Huntington's disease is a genetic disease, autosomal and dominant, that induces motor disorders, an inexorable deterioration of higher brain functions and psychiatric disturbances. At present, there are no known therapeutics against Huntington's disease. The Network of European CNS Transplantation and Restoration (NECTAR) has begun a program aimed at defining the conditions under which intrastriatal transplantation of fetal striatal cells could be attempted as an experimental treatment for Huntington's disease. This review presents the reasons why our group is considering participating in these trials. The validity of this therapeutic approach is supported by three main series of data: (i) neuropathological, clinical and imaging data indicate that Huntington's disease is, above all, a localized affection of a specific neuronal population ("medium spiny" neurons) in the striatum; (ii) a large body of experimental results, obtained in rats and non-human primates, demonstrates that transplanted fetal striatal cells are able to integrate the host brain and to substitute for previously lesioned host striatal neurons; (iii) expertise in clinical neural transplantation has now been acquired from the treatment of patients with Parkinson's disease. These different sets of data are presented and discussed in this review. There are a number of problems which do not yet appear to be entirely resolved, nor are they likely to be using the experimental models currently available. These problems are identified and explicitly presented as working hypotheses. (1) Anatomo-functional results obtained in rodents and non human primates with excitotoxic striatal lesions can serve as a basis for the extrapolation of what can be obtained from patients with Huntington's disease. (2). Huntington's disease can be efficiently fought by substituting degenerated striatal neurons alone. (3) Huntington's disease is due to a genetic defect which either hits the neurons that carry it directly or hits them indirectly only after several decades. Transplanted neurons, because they do not carry the gene or because they are of fetal origin, will not be rapidly affected by the ongoing disease process. Given the current state of knowledge, intracerebral transplantation appears to be the most serious opportunity (if not the only one that has been experimentally validated) for clinical improvement to be obtained in patients with Huntington's disease. The purpose of this review is to open a scientific discussion on its experimental bases before actual clinical trials start. PMID- 7477941 TI - Functional anatomy of the thalamic reticular nucleus as revealed with the [14C]deoxyglucose method following electrical stimulation and electrolytic lesion. AB - The [14C]-deoxyglucose quantitative autoradiographic method was used to map the metabolic changes induced by electrical stimulation and electrolytic lesion of the rostral pole of the thalamic reticular nucleus in the rat brain. Unilateral electrical stimulation of the thalamic reticular nucleus induced the following changes in glucose utilization: (i) local enhancement of metabolic activity within the stimulated thalamic reticular nucleus, (ii) increase in glucose consumption in the ipsilateral thalamic mediodorsal, centrolateral, ventromedial and ventrolateral nuclei, as well as in the nucleus accumbens, (iii) bilateral depression of metabolism in the locus coeruleus, periaqueductal gray, ventral tegmental area, and medial habenula, as well as contralateral metabolic depression in the substantia nigra reticulata, compacta and in the ventral pallidum. Unilateral electrolytic lesion of thalamic reticular nucleus elicited metabolic depression in the ipsilateral thalamic mediodorsal, centrolateral, ventrolateral and ventromedial nuclei, and metabolic activation in the dorsal tegmental nucleus bilaterally. The existence of a descending thalamic reticular nucleus input to the periaqueductal gray is supported by the depressed activity measured in brain stem structures after thalamic reticular nucleus stimulation. The similar effects observed in the periaqueductal gray and substantia nigra contralateral to the stimulated thalamic reticular nucleus indicate a possible flow of information from one thalamic reticular nucleus to the contralateral basal ganglia via the periaqueductal gray. The opposite effects induced in the dorsal thalamic nuclei by thalamic reticular nucleus stimulation and lesion support the gating role of the thalamic reticular nucleus in the information flow between thalamus and cortex. PMID- 7477942 TI - Functional anatomy of the thalamic centrolateral nucleus as revealed with the [14C]deoxyglucose method following electrical stimulation and electrolytic lesion. AB - The effects of electrical stimulation and electrolytic lesion of the thalamic intralaminar centrolateral nucleus were studied in the rat brain by means of the quantitative autoradiographic [14C]deoxyglucose method. Unilateral electrical stimulation of the centrolateral nucleus induced: (i) local increase in metabolic activity within the stimulated centrolateral nucleus and the ipsilateral thalamic mediodorsal nucleus, (ii) metabolic depression in all layers of the ipsilateral frontal cortex, (iii) bilateral increase in glucose consumption within the periaqueductal gray, pedunculopontine nucleus, and pontine reticular formation, and (iv) contralateral metabolic activation in the deep cerebellar nuclei. The unilateral electrolytic lesion of the thalamic centrolateral nucleus elicited metabolic depressions in several distal brain areas. The metabolic depression elicited in the mediodorsal, ventrolateral, and lateral thalamic nuclei, as well as in the caudate nucleus, the cingulate, and the superficial layers of forelimb cortex were ipsilateral to the lesioned side. The metabolic depression measured in the medulla and pons (medullary and pontine reticular formation, periaqueductal gray, locus coeruleus, dorsal tegmental, cuneiformis, raphe and pedunculopontine tegmental nuclei), the cerebellum (molecular and granular layers of the cerebellar cortex, interpositus and dentate nuclei), the mesencephalon (substantia nigra reticulata, ventral tegmental area and deep layers of the superior colliculus), the diencephalon (medial habenula, parafascicular, ventrobasal complex, centromedial and reticular thalamic nuclei), the rhinencephalon (dentate gyrus and septum), the basal ganglia (ventral pallidum, globus pallidus, entopeduncular and accumbens nuclei) and the cerebral cortex (superficial and deep layers of the frontal and parietal cortex, deep layers of the forelimb cortex) were bilateral. These functional effects are discussed in relation to known anatomical pathways. The bilateral effects induced by the centrolateral nucleus lesion reflect an important role of the centrolateral nucleus in the processing of reticular activating input and in the interhemispheric transfer of information. The cortical metabolic depression induced by centrolateral nucleus stimulation indicates the participation of this nucleus in attentional functions. PMID- 7477943 TI - Dependence of sensorimotor cortex neuron activity on noradrenergic and serotoninergic transmission in unspecific thalamic nuclei. AB - Background and evoked impulse activity of sensorimotor cortex neurons and conditioned reflex of trained cats were changed by blockers of noradrenaline and serotonin receptors applied through chemitrodes to thalamic complex of the centrum medianum and nucleus parafascicularis neurons. Application of obzidan, an antagonist of beta-adrenoreceptor to the complex of the centrum medianum and nucleus parafascicularis induced an initial increase and following decrease of the motor activity of cats. After obzidan application conditioned reflex remained unchanged at the initial, excitatory phase and started to decrease later. Lisergamide applied to the complex of the centrum medianum and nucleus parafuscicularis produced an initial decrease of movement activity and then drowsiness of the animals. Responses to conditioned stimuli were gradually decreased. The background impulse activity of the sensorimotor cortex neurons increased after obzidan application and decreased rapidly after lisergamide injection into the complex of the centrum medianum and nucleus parafascicularis. The responses of cortical neurons to conditioned stimuli were reduced, sometimes completely eliminated in both cases. Background and evoked impulse activity greatly increased by application of 5-hydroxytryptamine after preliminary lisergamide-induced depression of the impulse activity. It was concluded that under natural conditions the noradrenergic system exerts indirect inhibitor effects on the neocortex neurons through the centrum medianum and nucleus parafascicularis thalamic complex, whereas the serotoninergic system acting via the same complex increases the background and evoked impulse activity of the sensorimotor cortical neurons. PMID- 7477944 TI - Generators of somatosensory evoked potentials investigated by dipole tracing in the monkey. AB - Generators of somatosensory evoked potentials, elicited by electrical stimulation of the median nerve in anaesthetized monkeys (Macaca fuscata), were investigated by submitting a three-dimensional reconstructed brain model to dipole tracing, which can equate surface potential distributions to an approximate corresponding equivalent dipole. The following components of the somatosensory evoked potentials were simultaneously recorded from 21-27 epidural electrodes: P7 (the letter indicates positive or negative polarity; the number indicates the approximate latency of the peak in ms) was recorded widely from various locations on both the left and right hemispheres, P10 was recorded near the anterior side of the central sulcus contralateral to the stimulation side, N10 was recorded near the posterior side of the contralateral central sulcus, P12 was recorded on both sides of the contralateral central sulcus, and P18 was recorded posterior to the contralateral central sulcus. Current source generators (dipoles) of each component of somatosensory evoked potentials were localized by dipole tracing: a dipole for P7 was located in the thalamus contralateral to the stimulation side; a dipole for P10 and N10 in the posterior wall of the contralateral central sulcus (area 3b); a dipole for P12 in the contralateral post central gyrus (areas 1 and 2); and a dipole for P18 in the anterior wall of the contralateral intraparietal sulcus (area 5). The locations and latencies of dipoles that generated cortical components of somatosensory evoked potentials, estimated by dipole tracing, were confirmed by direct cortical surface recording from a 16-25 electrode array placed directly on the cortical surface; and multiple unit recording from the anterior and posterior parietal cortices. After excision of area 5, P18 and N18 were abolished, whereas P10, N10, and P12 were not affected. The results suggest that dipoles for somatosensory evoked potentials progressed from the thalamus to area 5 via the primary somatosensory area. This progress is consistent with the hierarchical sequence of somatosensory information processing. PMID- 7477945 TI - Neuronal responsiveness to various sensory stimuli, and associative learning in the rat amygdala. AB - Neuronal activities were recorded from the amygdala and amygdalostriatal transition area of behaving rats during discrimination of conditioned auditory, visual, olfactory, and somatosensory stimuli associated with positive and/or negative reinforcements. Neurons were also tested with taste solution and various sensory stimuli that were not associated with reinforcement. Of the 1195 neurons tested, 475 responded to one or more sensory stimuli. Of these, 256 neurons responded exclusively to a unimodal sensory stimulus, 128 to multimodal sensory stimuli, and the remaining 91 could not be classified. Distribution of unimodal neurons was correlated with anatomical projections to the amygdala from sensory thalamus or sensory cortices. Multimodal neurons were located mainly in the basolateral and central nuclei of the amgydala. Response latencies of neurons in the basolateral nucleus were longer than those in other nuclei and neurons in the central nucleus had both short and long latencies. Neurons responsive to a given stimulus were more frequently encountered in the amygdalas of the trained rats than in those of the rats not trained to associate that stimulus with a reinforcement. Multimodal neurons that responded to conditioned and/or unconditioned stimuli used in the associative learned tasks were concentrated in the basolateral and central nuclei. The results indicate that some amygdalar neurons receive exclusive single sensory information, and the others receive information from two or more sensory inputs. Considering the long latencies and multimodal responsiveness, the basolateral and central nuclei of the amygdala might be foci where various kinds of sensory information converge. It is also suggested that the basolateral and central nuclei of the amygdala have critical roles in associative learning to relate sensory information to reinforcement or affective significance. PMID- 7477946 TI - Cell-specific modulation of basal and seizure-induced neurotrophin expression by adrenalectomy. AB - Reports of glucocorticoid effects on neurotrophin expression suggest that adrenal hormones may contribute to the pattern of changes in the expression of these factors induced by neuronal activity and seizures. To examine this possibility, the present study evaluated the influence of adrenalectomy on basal expression and seizure-induced alterations in levels of nerve growth factor, brain-derived neurotrophic factor, and neurotrophin-3 messenger RNAs in hippocampus, entorhinal cortex, and superficial neocortex. For determination of hormone effects on basal expression, adult male rats were adrenalectomized and killed 10-14 days later with paired adrenal-intact controls. For studies of adrenal steroid involvement in expression following seizure, adrenalectomized and adrenal-intact rats received a seizure-producing lesion of the dentate gyrus hilus. Changes in neurotrophin messenger RNA content were assessed by quantitative in situ hybridization. Adrenalectomy alone had no significant effect on brain-derived neurotrophic factor messenger RNA content but did result in cell-specific decreases in nerve growth factor and neurotrophin-3 messenger RNAs. Nerve growth factor messenger RNA levels were reduced in hippocampal stratum granulosum, entorhinal cortex, and neocortex but not in cells of the hippocampal molecular layers or hilus. With adrenalectomy, neurotrophin-3 messenger RNA was virtually eliminated from CA2 stratum pyramidale, partially reduced in stratum granulosum, but unaffected in neurons of the hippocampal molecular layers or entorhinal cortex. These effects were partially reversed by corticosterone (2 mg/l) supplement to the drinking saline. In experimental-seizure rats, adrenalectomy did not alter the direction or basic pattern of seizure-induced changes in neurotrophin expression but did change the time courses and magnitudes of these effects. In all areas measured, brain-derived neurotrophic factor messenger RNA content was more greatly and persistently elevated by seizure in adrenalectomized as compared with adrenal-intact rats. In contrast, with adrenalectomy seizures induced smaller increases in nerve growth factor messenger RNA content. Adrenalectomy augmented the decrease in neurotrophin-3 messenger RNA induced by seizure in hippocampus but not in entorhinal cortex. These results demonstrate that adrenal hormones play a major role in the regulation of basal nerve growth factor and neurotrophin-3 messenger RNA expression by specific populations of forebrain neurons. Moreover, the adrenal steroids have opposite effects on activity-dependent changes in brain-derived neurotrophic factor and nerve growth factor messenger RNA levels but are not required for the basic pattern of changes in neurotrophin messenger RNA expression elicited by recurrent seizures. PMID- 7477947 TI - Hippocampal homosynaptic long-term depression/depotentiation induced by adrenal steroids. AB - The effects of adrenal steroids on synaptic plasticity were investigated in the dentate gyrus of the hippocampus. Experiments were performed in either adrenalectomized or intact (non-adrenalectomized), anesthetized rats. High frequency stimulation was applied to the medial perforant pathway at three different frequencies; 100, 200 or 400 Hz, either post- or pre- and post administration of the specific Type-II adrenal steroid receptor agonist RU 28362. High-frequency stimulation prior to RU 28362 administration produced a frequency dependent long-term potentiation of the population spike, with 100 Hz showing no long-term potentiation and 400 Hz the highest degree of potentiation. In contrast, following administration of RU 28362, high-frequency stimulation produced a long-term depression (in comparison to baseline). In the experiments in which high-frequency stimulation was applied both pre- and post-RU 28362 administration, the size of the population spike was initially potentiated and then depotentiated after the RU 28362 injection. This effect was also frequency dependent, although opposite to the long-term potentiation effect. That is, 400 Hz was the least effective frequency for inducing long-term depression/depotentiation, while 100 Hz was the most effective. Long-term depression/depotentiation was immediate following high-frequency stimulation and lasted for the extent of the recording session, in some cases longer than 1 h. Similar to the finding reported in the accompanying paper, induction of long-term potentiation was substantially suppressed by RU 28362. However, in a number of experiments long-term potentiation could still be induced after RU 28362 administration, even after long-term depression/depotentiation had been established. In these cases, stimulation at the higher frequencies was necessary.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7477948 TI - Opposing roles of type I and type II adrenal steroid receptors in hippocampal long-term potentiation. AB - Studies were performed in vivo on the dentate gyrus to investigate the possible involvement of Type I and Type II adrenal steroid receptors in the mediation of reported adrenal steroid effects on long-term potentiation, through the use of specific Type I and Type II receptor agonists and antagonists. In adrenalectomized rats, administration of aldosterone, a specific Type I agonist, produced a marked enhancement in long-term potentiation, in comparison to either the adrenalectomized or sham adrenalectomized controls. Administration of RU 28318, a Type I antagonist, which by itself had minimal effects, blocked the aldosterone enhancement. In contrast, administration of the specific Type II agonist, RU 28362, produced a marked decrement in the induction of long-term potentiation. The RU 28362 effect was blocked by a prior injection of the Type II antagonist, RU 38486. Neither adrenalectomy nor administration of any of the steroid agonists or antagonists had noticeable effects on neuronal excitability (as determined by the field potentials), nor on post-tetanic potentiation. These findings are consistent with other studies that have shown a biphasic effect of increasing levels of corticosterone on long-term or prime burst potentiation. Taken together, these studies suggest that Type I receptors, with a high affinity for corticosterone, and Type II receptors, having a lower affinity for corticosterone, form a two-level recognition system to modulate induced synaptic plasticity in opposite directions in the dentate gyrus and possibly also in Ammon's horn. PMID- 7477949 TI - Temperature change-induced potentiation: a comparative study of facilitatory mechanisms in aged and young rat hippocampal slices. AB - The effect of temperature changes in the medium on the evoked potentials of pyramidal neurons in response to the electrical stimulation of Schaffer collaterals was investigated in both young (one to two months) and aged (24-28 months) rat hippocampal slices. Temperature increase was shown to reduce, and subsequent temperature decrease to enhance, the population spike amplitude in both young and aged rats. Temperature decrease produced a long-lasting (> 30 min) and important potentiation (180%) of population spike amplitude in young but not in aged rats. Long-term post-tetanic potentiation was maintained in young but suppressed in aged rats by temperature changes. The impairment of temperature induced potentiation in aged rats is suggested as a tool for studying promnesic drugs. PMID- 7477950 TI - Long-term and regional specific changes in [3H]flunitrazepam binding in kindled rat hippocampus. AB - The binding of the GABAA/benzodiazepine receptor agonist [3H]flunitrazepam was studied in the hippocampus of rats kindled by daily stimulation of the Schaffer collaterals, using semi-quantitative autoradiography. Two kindled stages were investigated: (i) 24 h after the last generalized tonic-clonic seizure (fully kindled) and (ii) 28 days after the last generalized seizure (long-term). The binding of [3H]flunitrazepam was determined at two concentrations, 3 and 16 nM. In the CA1 area, we found a small but significant decrease (ca. 10%), both in the 3 and 16 nM [3H]flunitrazepam binding at the fully kindled stage. In contrast, there was a significant increase in the 3 nM binding (c. 15%) at the long-term stage. The 16 nM binding was not significantly different from control binding at this stage. In the granular and molecular layers of the fascia dentata, we found at both kindled stages a significantly increased 3 nM (ca. 9 and 19%, respectively) and 16 nM (ca. 19 and 14%, respectively) binding. Furthermore, we found that muscimol was still able to enhance the [3H]flunitrazepam binding in kindled animals, indicating that the GABAA receptor agonist binding site and benzodiazepine agonist binding site are still functionally coupled. The changes in [3H]flunitrazepam binding at the fully kindled stage are in agreement with the recently observed kindling-induced changes in [3H]muscimol binding in the hippocampal formation of the same animals [Titulaer M. N. G. et al. (1994) Neuroscience 59, 817-826] and extend these observations to the benzodiazepine modulatory site of the GABAA receptor.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7477951 TI - Pentylenetetrazol causes polysynaptic responses to appear in the dentate gyrus. AB - In a particular brain region specific changes in inhibition or excitation may be the basis of seizure initiation. Alternatively, changes in the balance of excitation and inhibition in the circuit, which may be detectable as polysynaptic responses may be more important indicators of epileptogenesis. That the appearance of polysynaptic responses precedes the initiation and, therefore, may be necessary for the onset of epileptiform activity in the hippocampal parahippocampal circuit was tested using the chemical convulsant pentylenetetrazol. Excitation and paired-pulse inhibition were measured in CA1 and the dentate gyrus of the urethane-anaesthetized rat before and after administration of pentylenetetrazol. In addition, three polysynaptic responses were monitored. In both CA1 and the dentate gyrus, pentylenetetrazol, 100 mg/kg, caused a trend towards increased excitability and caused a relatively mild loss of inhibition. Two polysynaptic responses appeared in the dentate gyrus after the administration of pentylenetratrazol, both apparently mediated through the entorhinal cortex. A polysynaptic response of the CA1 pyramidal neurons to contralateral angular bundle stimulation was not observed. These experiments demonstrate that pentylenetetrazol will facilitate only the appearance of polysynaptic responses mediated through the entorhinal cortex. These results support the hypothesis that pentylenetetrazol has a specific action within the entorhinal cortex that may facilitate the synchronization and spread of epileptiform activity. These results are also consistent with the hypothesis that the appearance of polysynaptic responses may be necessary for the onset of epileptogenesis in the hippocampal-parahippocampal circuit. PMID- 7477953 TI - (1S,3R)-1-aminocyclopentane-1,3-dicarboxylic acid-induced burst firing is mediated by a native pertussis toxin-sensitive metabotropic receptor at rat dorsolateral septal nucleus neurons. AB - We have reported previously that a selective metabotropic glutamate receptor agonist, (1S,3R)-1-aminocyclopentane-1,3-dicarboxylic acid (1S,3R-ACPD), caused two primary postsynaptic membrane changes, namely, a slow membrane depolarization, and burst firing in rat dorsolateral septal nucleus neurons. In addition, (1S,3R)-1-aminocyclopentane-1,3-dicarboxylic acid also potentiates a slow after depolarization in rat dorsolateral septal nucleus neurons. We now report that, among all the postsynaptic membrane changes induced by (1S,3R)-1 aminocyclopentane-1,3-dicarboxylic acid, only the burst firing was selectively blocked by pertussis toxin pretreatment. Thus, aminocyclopentane-1,3-dicarboxylic acid induced burst firing was mediated by a metabotropic receptor coupled to a pertussis toxin-sensitive GTP-binding protein, while the other induced cellular responses may be mediated by metabotropic glutamate receptors insensitive to pertussis toxin. We further characterized this receptor pharmacologically. This metabotropic receptor is activated by several metabotropic glutamate receptor agonists, but is insensitive to L-glutamate or L-aspartate. On the basis of its agonist activity profile, particularly the ineffectiveness of glutamate as an agonist, we have tentatively assigned the name aminocyclopentane-1,3-dicarboxylic acid metabotropic receptor, to this native, pertussis toxin-sensitive metabotropic receptor in the dorsolateral septal nucleus. Furthermore, this receptor is coupled to protein kinase C, probably via a phospholipase C independent pathway. PMID- 7477952 TI - Decreased GABA release following tonic-clonic seizures is associated with an increase in extracellular glutamate in rat hippocampus in vivo. AB - The effects of maximal electroshock, used as a model of generalized seizures, were studied on extracellular GABA and glutamate levels in the ventral hippocampus of the freely-moving rat, using in vivo microdialysis. Following a maximal electroshock there was a rapid decline in GABA levels (46 +/- 5%) in the 20 min immediately after the seizure and levels remained depressed for a further 60 min. However, although there was a transient small decrease (11 +/- 2%) in glutamate levels in the first 20 min post-ictally, there followed a more prolonged, larger increase in the next 40 min. Maximal electroshock, administered in the absence of extracellular calcium, did not change GABA levels, while glutamate levels were again increased (42 +/- 8%) in the 40-80 min after the shock. Local perfusion with nickel (1 mM) to block T-type calcium channels had no effect on basal GABA or glutamate levels but prevented maximal electroshock induced changes in both amino acids. Experiments were carried out to test the hypothesis that the post-ictal increased glutamate release was due to the decrease in GABA release. Perfusion with the potent GABA re-uptake inhibitor NNC 711, for 60 min prior to administration of maximal electroshock, increased GABA levels (436 +/- 58%) and abolished the seizure-induced decrease. Basal glutamate levels were not affected by perfusion with NNC-711 but subsequent maximal electroshock also failed to affect levels. Local perfusion with the GABAA receptor antagonist bicuculline (1, 10 and 100 microM) had no effect on basal GABA levels but glutamate levels were increased (46 +/- 5%) after perfusion with 100 microM bicuculline.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7477954 TI - Differential distribution of AMPA-selective glutamate receptor subunits in the parabrachial nucleus of the rat. AB - The parabrachial complex is made up of at least 11 cytoarchitectonically distinct subnuclei which differ in their anatomical connections and neurotransmitter content, as well as the functions they subserve. To determine whether parabrachial subnuclei also express different types of glutamate receptors, we undertook a light microscopic examination of the regional distribution of alpha amino-3-hydroxy-5-methyl-4-isoxazolepropionate receptor subunits within the parabrachial complex using antibodies directed against synthetic peptides corresponding to the C-terminal parts of alpha-amino-3-hydroxy-5-methyl-4 isoxazolepropionate receptor subunits. Antibodies that recognize GluR1 subunits stained cells mainly in the central lateral parabrachial subnucleus, whereas GluR4 antibodies selectively stained cells in the internal lateral subnucleus. In contrast, antibodies directed against the GLuR2/3 subunits stained neurons in every parabrachial subnucleus, although the most dense labelling was seen in the external lateral cell group. These differences in expression of alpha-amino-3 hydroxy-5-methyl-4- isoxazolepropionate receptor subtypes may eventually allow selective activation or inhibition of specific subsets of neurons in the parabrachial complex. PMID- 7477955 TI - Time-dependent inhibition of hindlimb somatic afferent transmission within nucleus tractus solitarius: an in vivo intracellular recording study. AB - In a previous study we demonstrated that hindlimb somatic afferent stimulation evokes excitatory responses from neurons in nucleus tractus solitarius. When paired electrical stimuli were delivered to hindlimb somatic afferents, the unit response to the second stimulus was significantly reduced compared with responses to the first. This temporal response pattern has been termed time-dependent inhibition since responses to the second stimulus recover as the interval separating the first and second stimuli is increased. To examine possible synaptic mechanisms for somatic afferent-evoked time-dependent inhibition, intracellular recordings were made from nucleus tractus solitarius neurons in anesthetized, paralysed rats. Skeletal muscle afferent fibers were activated by electrically stimulating the right tibial nerve in the hindlimb and neuronal responses recorded in the contralateral nucleus of the solitary tract. Time dependent inhibition of tibial nerve-evoked unit discharge was studied using a conditioning-test stimulation procedure, with the first (conditioning) and second (test) stimuli separated by intervals of 50, 150 and 250 ms. In 49 units that responded to tibial nerve stimulation, 46 were excited and three were inhibited. Among units excited, 25 displayed a unimodal response that had an onset latency of 21.3 +/- 5.9 ms. The remaining 21 units responded with a bimodal discharge pattern characterized by both a short-latency and a long-latency response. The onset latency of the early response was 23.7 +/- 5.3 ms and was not statistically different from the unimodal response onset latency. The onset latency of the late response was 143 +/- 23.9 ms.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7477956 TI - Low nanomolar serotonin inhibits the glutamate receptor/nitric oxide/cyclic GMP pathway in slices from adult rat cerebellum. AB - The function of serotonin afferents to the cerebellum has been investigated by monitoring the effects of serotoninergic drugs on the production of cyclic GMP elicited in cerebellar slices by activation of ionotropic glutamate receptors. Exposure of adult rat cerebellar slices to N-methyl-D-aspartate (1 nM to 1 microM) or to (RS)-alpha-amino-3-hydroxy-5-methyl-4-isoxazole propionate (AMPA; 1 nM to 10 microM) elicited concentration-dependent and saturable rises in the levels of cyclic GMP. These responses were blocked by selective antagonists at the N-methyl-D-aspartate or AMPA receptors and by inhibiting nitric oxide synthase, but were insensitive to tetrodotoxin. When tested between 0.1 and 10 nM, serotonin, the serotonin1A receptor agonist (+/-)-8-hydroxy-2-(di-n propylamino)tetralin and the serotonin2 receptor agonist (+/-)-1-(2,5-dimethoxy-4 iodophenyl)-2-aminopropane inhibited, concentration-dependently, the cyclic GMP responses evoked by near-maximal (0.1 microM) concentrations of N-methyl-D aspartate or AMPA. The EC50 values (concentrations causing half-maximal effect) ranged between 0.7 and 2.1 nM. The actions of serotonin were totally abolished by methiothepin, a mixed-type serotonin receptor antagonist. Thus, the serotonergic cerebellar afferents may exert a potent inhibitory control on the excitatory transmission mediated by N-methyl-D-aspartate and AMPA receptors; the inhibition occurs through both serotonin1A and serotonin2 receptors. As the glutamate receptor-dependent cyclic GMP responses involve production of nitric oxide, a diffusible activator of guanylate cyclase, the above inhibitory serotonin receptors may have multiple localization.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7477957 TI - The involvement of multiple calcium channel sub-types in glutamate release from cerebellar granule cells and its modulation by GABAB receptor activation. AB - In this study, we have examined both the ability of various Ca2+ channel sub types to support the release of [3H]glutamate from cerebellar granule neurons and the mechanism of action involved in the modulation of glutamate release by the GABAB receptor agonist, (-)-baclofen. Cerebellar granule neurons were stimulated to release newly synthesized [3H]glutamate by K(+)-evoked depolarization. Stimulated release was entirely calcium-dependent and abolished by the presence of 200 microM cadmium. Release of glutamate was not affected by either tetrodotoxin or 5-aminophosphonovaleric acid but was potentiated by dihydrokainate and inhibited by 6-cyano-7-nitroquinoxaline-2,3-dione. Stimulated glutamate release was partially inhibited by both the L-type calcium channel blocker, nicardipine, and the N-type calcium channel blocker, omega-conotoxin GVIA; however, the P/Q-type calcium channel blocker omega-agatoxin IVA inhibited release of glutamate only after pre-incubation of cells with omega-conotoxin GVIA. K(+)-stimulated release of glutamate was observed when stimulated either in the presence of Ca2+ or of Ba2+ and similar inhibition of release by (-)-baclofen was seen under both conditions. In contrast to these results, ionomycin-evoked glutamate release was greatly reduced as compared to K(+)-evoked release and was not modulated by (-)-baclofen. In the presence of omega-conotoxin GVIA alone, inhibition of release by (-)-baclofen was attenuated but not abolished. Following block of nicardipine-sensitive channels, inhibition of release by (-)-baclofen was still present, and after prior block of omega-conotoxin GVIA-sensitive channels the presence of nicardipine restored the ability of (-)-baclofen to inhibit residual release of glutamate. Modulation of glutamate release by (-) baclofen was unaffected by the presence of omega-agatoxin IVA alone; however, after block of both omega-conotoxin GVIA- and omega-agatoxin IVA-sensitive channels, inhibition of release by (-)-baclofen was completely abolished. These results indicate that multiple sub-types of voltage-dependent calcium channels are present on the presynaptic terminals of cerebellar granule neurons and support K(+)-stimulated release of [3H]glutamate. Modulation of release by GABAB receptor activation appears to be dependent upon interaction of this receptor with a number of voltage-sensitive calcium channels, including omega-conotoxin GVIA-sensitive and omega-agatoxin IVA-sensitive channels. PMID- 7477958 TI - Alpha-1 subunits of voltage gated Ca2+ channels in the mesencephalon x neuroblastoma hybrid cell line MES23.5. AB - The identity of alpha 1 subunits from voltage operated Ca2+ channels was determined in the rat/mouse mesencephalon x N18TG2 hybridoma cell line MES23.5, by sequence analysis of reverse transcription-polymerase chain reaction products and antagonist binding. Sequences were derived from the L-(alpha 1D), Q-(alpha 1A) and omega-conotoxin GVIA sensitive N-type (alpha 1B) Ca2+ channel alpha 1 subunits. The amplified fragments, which are homologous to the region between domain III and IV of known alpha 1 subunits, reveal splice variation in the L- and Q-type alpha 1 subunit of MES23.5 cells. The transcripts of alpha 1 subunits in these cells were quantified by RNAase protection assay. The data show the existence of different Ca2+ channel types in a single cell line and may reflect multiple functions of voltage operated Ca2+ channels during growth, differentiation and transmitter release. PMID- 7477959 TI - Evidence for a sodium-dependent potassium conductance in frog myelinated axon. AB - After blockade of the voltage-dependent potassium conductances by intracellular application of 4-aminopyridine and tetraethylammonium in frog myelinated axons, a set of brief (0.1 ms) intracellular depolarizing pulses or a long (200 ms) depolarizing pulse evoked a train of action potentials. Under both experimental conditions a hyperpolarizing afterpotential appeared (duration 367 ms +/- 34, mean +/- S.E., n = 15). The purpose of this study was to investigate the properties of this hyperpolarizing afterpotential. It was found that the hyperpolarizing afterpotential increases in amplitude with: (1) the number of sodium-dependent action potentials; (2) action potential broadening (following potassium channels blockade); and (3) the level of depolarization during a current step. Application of tetrodotoxin prevented the activation of the hyperpolarizing afterpotential by any of the above stimuli. The hyperpolarizing afterpotential was unaffected by: (1) 8-acetyl-strophanthidin, an agent that poisons the electrogenic pumping in the axon; (2) blocking calcium influx with extracellular 10 mM magnesium or 2 mM manganese; and (3) buffering of the intracellular calcium, using EGTA in the recording microelectrode. Extracellular application of tetraethylammonium, but not 4-aminopyridine, reduced the hyperpolarizing afterpotential. The hyperpolarizing afterpotential reversed at >> -92 mV. Increasing the external potassium concentration from 2 to 10 mM shifted the reversal potential +14.5 mV, indicating that the hyperpolarizing afterpotential is a potassium mediated conductance.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7477960 TI - Different voltage-dependent potassium conductances regulate action potential repolarization and excitability in frog myelinated axon. AB - Intracellular microelectrode recordings were used to examine the effects of the potassium channel blockers: 4-aminopyridine, a selective blocker of fast potassium conductances gKf1 and gKf2,13 and tetraethylammonium, a blocker of gKf1, gKf2 and the slow conductance gKs,13 on the repetitive activity of large myelinated axons of frog. The blockers were applied intracellularly by diffusional leak of the agents from the recording microelectrode containing either 4-aminopyridine or a mixture of 4-aminopyridine and tetraethylammonium. A decrease in outward rectification, a measure of the block of the potassium conductances, was evident within 5 min of axon impalement. Within 30 min 80% of maximal blockade was observed during prolonged recording sessions (> 1 h). Parallel with the resistance increase, the action potential duration increased (up to 5 ms). This was attributed to the block of gKf2. The excitability regularly increased, manifested as a train of action potentials (a decrease in accommodation) for a maximum of 200 ms (54 +/- 8 vs 111 +/- 22, 4-aminopyridine vs 4-aminopyridine-tetraethylammonium, respectively, n = 8 and 6, P < 0.006). The presence of 4-aminopyridine-tetraethylammonium in the microelectrodes decreased the spike frequency adaptation (the instantaneous action potential frequency per spike interval number) observed in fibres treated with 4-aminopyridine alone (32 +/- 9 vs 7 +/- 1 Hz; 4-aminopyridine vs 4-aminopyridine-tetraethylammonium, n = 8 and 6, P < 0.04). This effect was attributed to block of gKs by the tetraethylammonium.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7477961 TI - Neonatal 6-hydroxydopamine lesions lead to opposing changes in the levels of dopamine receptors and their messenger RNAs. AB - Previous studies have established that selective damage to the early-developing components of the mesostriatal dopamine system produces profound changes in dopamine D1 receptor-mediated behaviors, while decreasing D1 receptor density. In order to better understand the effects of early intrastriatal 6-hydroxydopamine lesions, we studied the ontogenetic expression (postnatal days 7, 14, 35 and 90) of D1 and D2 receptors, and their corresponding messenger RNAs, in rats that had received intrastriatal 6-hydroxydopamine or vehicle lesions on postnatal day 1. Using receptor autoradiography, significant (P < 0.05) decreases in [3H]SCH 23390 binding to D1 receptors in the rostral and caudal dorsomedial and ventromedial caudate of 6-hydroxydopamine-lesioned animals were evident by postnatal day 7, and remained depressed at all future time points. A significant decrease in D1 receptor concentration occurred in the dorsolateral caudate at later time points (postnatal days 35 and 90). [3H]Spiperone binding to D2 receptor sites was unchanged throughout the entire study. In situ hybridization for D1 and D2 messenger RNA expression showed contrasting results. 6-Hydroxydopamine induced significant decreases of D1 messenger RNA levels in the dorsolateral and dorsomedial caudate by postnatal day 7. By postnatal day 14, messenger RNA expression was significantly elevated in the dorsomedial and ventromedial caudate of the 6-hydroxydopamine group, and remained elevated thereafter. D1 messenger RNA levels became elevated in the lateral caudate at later time points (postnatal days 35 and 90). The opposing changes in D1 receptor concentrations and the messenger RNA encoding the protein did not occur as a consequence of increased transport of D1 receptors to striatonigral terminals. D2 messenger RNA levels in the dorsal caudate were significantly decreased on postnatal day 7, and became higher than controls at postnatal day 14, but were unchanged from controls at later time points. Together, the D1 receptor and D1 messenger RNA findings suggest that the normal relationship between levels of D1 receptor transcript and D1 receptor protein is permanently altered following the early loss of dopamine. In contrast, the results indicate that dopamine plays a minor role in the early postnatal development of the D2 receptor protein and transcript. These findings suggest that dopamine may be involved in the coordinated expression of some dopamine receptors and their corresponding messenger RNAs during development. PMID- 7477962 TI - Axonal and dendritic transport of internalized neurotensin in rat mesostriatal dopaminergic neurons. AB - Previous studies have demonstrated that neurotensin is internalized and retrogradely transported in neurons of the substantia nigra following its intracerebral injection in the neostriatum. The aim of the present study was to compare the intracellular distribution of retrogradely transported material with that observed following internalization of the peptide at the somatodendritic level and to confirm that the internalization was confined to dopamine neurons. To document somatodendritic internalization, slices (350 microns) from the rat ventral midbrain were incubated in vitro with 20 mM fluoresceinylated neurotensin, a fluorescent derivative of neurotensin, and immunostained 5-60 min later for tyrosine hydroxylase. To document retrograde transport, rats were injected with the same compound into the neostriatum and the brains processed for tyrosine hydroxylase immunohistochemistry 4.5 and 8 h later. Confocal laser microscopic examination of superfused slices revealed that fluoresceinylated neurotensin was internalized at the level of the perikarya and processes of neurons in the substantia nigra, ventral tegmental area and interfascicular nucleus. At short time intervals, the label was detected in the form of small, intensely fluorescent particles distributed within the cytoplasm of both perikarya and dendrites. At longer time intervals, these fluorescent particles were larger, less numerous and confined to the perikarya where they eventually clustered against the nucleus. Following intrastriatal injection of fluoresceinylated neurotensin, retrogradely labeled cells were apparent throughout the substantia nigra, pars compacta, as well as in the lateral part of the ventral tegmental area. Here again, the label took the form of small fluorescent particles, comparable in size, shape and distribution to those detected following superfusion of midbrain slices. In both labeling conditions, fluoresceinylated neurotensin was almost exclusively confined to tyrosine hydroxylase-immunoreactive cells. These results indicate that neurotensin is internalized throughout the terminal and dendritic arborization of mesostriatal dopamine cells and that the internalized peptide is transported centripetally from both locations to the soma of the cells. The clustering of fluorescent particles in the perinuclear region of the cells further suggests that the internalized process may play a role in the long term transcellular signalling. PMID- 7477963 TI - Nicotine treatment counteracts perinatal asphyxia-induced changes in the mesostriatal/limbic dopamine systems and in motor behaviour in the four-week-old male rat. AB - In the present study, the effects of nicotine treatment on the changes induced by perinatal asphyxia in exploratory and D-amphetamine-induced behaviour, and in the number of brain tyrosine hydroxylase-immunoreactive nerve cell bodies were investigated in four-week-old male rats. Asphyxia was induced in pups by placing the fetuses, still in their uterus horns removed by hysterectomy from full-term pregnant rats, in a 37 degrees C water bath for 15-16 min or 19-20 min. Surviving male pups were treated with nicotine via suckling from surrogate mothers implanted subcutaneously with Alzet minipumps containing nicotine (0.2 mumol/kg per h) for four weeks. The minipumps implanted in the mothers of sham-treated animals contained saline only. After treatment, exploratory behaviour and D amphetamine-induced behaviour was analysed in a computerized "activity" box. After the behavioural experiments, the rats were taken for tyrosine hydroxylase immunohistochemistry, and the total number of tyrosine hydroxylase immunoreactive cell bodies were counted in the A9 and A10 regions of the substantia nigra and the ventral tegmental area, respectively. Nicotine serum levels were measured using gas chromatography in selected asphyctic and control pups at different periods after delivery. During the exploratory phase, in saline-nurtured rats, 15 16 min of asphyxia slightly increased (approximately 25%) locomotion, motility and rearing. In contrast, 19-20 min of asphyxia reduced the locomotion and rearing by approximately 50%, as compared to controls. An increase in amphetamine induced behaviours was observed after 15-16 min, but not after 19-20 min of asphyxia, as compared to controls.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7477964 TI - Quantitative analysis of the variability of substantia nigra pigmented cell clusters in the human. AB - At present there is no consensus concerning the internal organization of the human substantia nigra, despite its pivotal role in neurodegenerative conditions. We have quantitatively analysed the variability in the pattern of clusters of melanin-pigmented neurons in the human substantia nigra using serial section analysis and computer reconstructions. The substantia nigra pars compacta showed a bilaminar organization consisting of the pars medialis and pars lateralis, as well as dorsal and ventral tiers as described previously [D. A. McRitchie et al. (1995) J. comp. Neurol. (in press)]. Both the dorsal and ventral tiers could be further subdivided into three mediolateral cell columns based on position and cell density. The presence and arrangement of these cell clusters was most variable in transverse sections (the plane currently used for diagnostic neuropathology). Quantitative assessment of the topographical pattern of cell loss within single transverse sections of the human substantia nigra should therefore be treated with some caution. In contrast, the full rostrocaudal extent of the cell columns could be seen in horizontal sections. Thus, consistent samples of larger numbers of pigmented neurons per region were found in this section plane, although only two cell columns were found in most sections. Our results show that greater quantitative reliability can be achieved with horizontal sections of the substantia nigra. PMID- 7477965 TI - Involvement of the dorsal raphe but not median raphe nucleus in morphine-induced increases in serotonin release in the rat forebrain. AB - In vivo microdialysis was used to determine if morphine produces increases in extracellular serotonin in specific brain sites. With citalopram included in the dialysis solution to block reuptake, serotonin was measured in 11 brain sites of unanesthetized rats. After systemic morphine (10 mg/kg, s.c.), increases in extracellular serotonin were observed in the nucleus accumbens, amygdala, frontal cortex, striatum, thalamus, hypothalamus and ventral hippocampus. These seven areas are innervated either by dorsal raphe nucleus projections alone, or by projections from both dorsal and median raphe nucleus. In contrast, serotonin was not significantly increased in the medial septal nucleus and dorsal hippocampus. These two areas are selectively innervated by projections from the median raphe nucleus. After systemic morphine, serotonin was increased in the dorsal raphe nucleus, but not in the medial raphe nucleus. Local infusion of morphine through a microdialysis probe in the dorsal raphe nucleus induced a dose-dependent increase of serotonin in the nucleus accumbens, but not in the medial septum. In contrast, infusion of morphine into the median raphe nucleus had little effect on serotonin in either the nucleus accumbens or septum. Infusion of morphine into either the dorsal or median raphe nucleus elicited increased behavioral activity and hyperthermia. These data provide evidence that morphine acts in the area of the dorsal raphe nucleus, but not the median raphe nucleus, to enhance serotonin release in specific forebrain sites, and that the increases in serotonin in the dorsal raphe nucleus projection sites are not an indirect effect of changes in behavioral state or body temperature. PMID- 7477966 TI - Central and peripheral expression of galanin in response to inflammation. AB - Using in situ hybridization, immunohistochemistry and receptor binding methodology, the galanin messenger RNA levels, galanin binding and galanin-like immunoreactivity were examined in rats injected with carrageenan into the left hindpaw. Three days after injection, a distinct increase (63%) in galanin messenger RNA-positive neurons was observed in the medial laminae I and II of the ipsilateral dorsal horn (lumbar 4 and 5) as compared to the contralateral side. However, no alteration was found in galanin binding and galanin-like immunoreactivity in the dorsal horn. In dorsal root ganglia (lumbar 5), inflammation induced a significant decrease in galanin messenger RNA (39%) and galanin peptide (47%) on the ipsilateral side. Galanin binding was not detected in dorsal root ganglia, neither on the inflammatory nor on the control side. Increased levels of galanin-like immunoreactivity and galanin messenger RNA were seen in cells in the inflamed dermis and epidermis, especially in stratum granulosum. Most of the galanin-immunoreactive cells contained ED1-like immunoreactivity, a marker for macrophages. A strong galanin binding was seen in the inflamed dermis. Such binding sites may be targets for galanin released from local cells in inflamed dermis. Taken together, our results suggest that both neuronal and non-neuronal galanin or a galanin-like peptide is involved in the response to inflammation. PMID- 7477967 TI - Characteristics of substance P-evoked release of amino acids from neonatal rat spinal cord. AB - The characteristics of release of amino acids evoked by substance P from the neonatal rat spinal cord were examined. A hemisected spinal cord was continuously perfused and the release of amino acids into the perfusate was measured using high-performance liquid chromatography with a precolumn derivatization technique. Substance P (10 microM) evoked a significant increase in the release of aspartate, glutamate, GABA, glycine and taurine. The substance P-evoked release of these five amino acids was not reduced by Ca(2+)-free medium, but was blocked by [D-Pro4,D-Trp] substance P4-11 (10 microM). Perfusion of the spinal cord with low-Na+ medium (22 mM) induced a marked increase in the high-K+ (90mM)-evoked release of GABA, glutamate and glycine. In contrast, the substance P-evoked release of the five amino acids was significantly decreased by the low-Na+ medium. Similarly, perfusion of the spinal cord with low-Cl- medium (8 mM) increased the high-K(+)-evoked release of GABA and glycine, but decreased the substance P-evoked release of GABA, glycine and taurine. The substance P-evoked release of the five amino acids was dose-dependently blocked by d-tubocurarine (3 10 microM), whereas it was not blocked by tetrodotoxin (0.2 microM) or amiloride (30 microM). Compound 48/80 (10 micrograms/ml), a histamine-releasing agent on mast cells, evoked release of the amino acids from the spinal cord with characteristics similar to those of substance P-evoked amino acid release.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7477968 TI - Neurotoxicity of acromelic acid in cultured neurons from rat spinal cord. AB - Acromelic acid A, which contains the kainic acid structure in its molecule, is known to cause selective damage of interneurons in the rat lower spinal cord. In the present study, the potent neurotoxicity of acromelic acid A was demonstrated in cultured rat spinal neurons in terms of the activity of lactate dehydrogenase that was released from degenerated neurons into the culture medium. Acromelic acid A increased the lactate dehydrogenase activity in time- and concentration dependent manners, and its EC50 was about 2.5 microM, which was much lower than that of kainic acid (70 microM) and (RS)-alpha-amino-3-hydroxy-5-methyl-4 isoxazolepropionic acid (EC50; 11 microM). The maximum level of lactate dehydrogenase released by acromelic acid A was quite similar to that by kainic acid, but was about twice the level produced by (RS)-alpha-3-hydroxy-5-methyl-4 isoxazolepropionic acid. Exposure to acromelic acid A caused release of L glutamate from the cells into the medium; however, the concentration of L glutamate released was far below the level for inducing the neurotoxic effects. The neurotoxicity of 10 microM acromelic acid A was almost completely inhibited by 30 microM 6-nitro-7-sulphamoylbenzo(F)quinoxaline-2,3-dione and 6-cyano-7 nitroquinoxaline-2,3-dione, potent antagonists for non-N-methyl-D-aspartate receptors, but was partly (35%) reduced by 30 microM dizocilpine maleate. In cultured hippocampal neurons, the toxicity of acromelic acid A was significantly less effective (EC50: 18 microM) than that in spinal neurons, whereas the toxicity of kainic acid was almost the same in both neurons. These results suggest that acromelic acid A directly activates non-N-methyl-D-aspartate receptors on the cultured spinal neurons to induce neuronal death. A new type of non-N-methyl-D-aspartate receptors which is specific to acromelic acid A is suggested to be present at least in spinal neurons. PMID- 7477969 TI - Cytochrome P450 and associated monooxygenase activities in the rat and human spinal cord: induction, immunological characterization and immunocytochemical localization. AB - We have discovered cytochrome P450 and associated monooxygenase activities in microsomes prepared from spinal cord tissues from rats and a human. Cytochrome P450 levels and nicotinamide adenine dinucleotide phosphate cytochrome c reductase activities in microsomes from rat spinal cord were similar to those observed from the whole brain. However, certain monooxygenase activities were significantly lower in the rat spinal cord microsomes as compared to the corresponding activities observed in the whole brain. Cytochrome P450-mediated monooxygenase activities were also detectable in microsomes prepared from human spinal cord. Immunoblot analyses of rat and human spinal cord microsomes using antisera to various forms of hepatic cytochrome P450 namely (2B1 + 2B2), 1A1, 1A2 and 2E1 revealed the presence of immunologically similar forms. The spinal cord microsomes also cross-reacted with the antiserum to the phenobarbital-inducible form of rat brain cytochrome P450. Immunocytochemical stain was predominant in the gray horns of the rat spinal cord. At the cervical level, lamina 1 and 2 representing the substantia gelatinosa were intensely stained. In the ventral horns, lamina 7, 8 and 9 containing the large motor neurons were strongly labelled, while small neurons revealed variable staining. In the white matter, the glial cells were stained but the axons remained non-reactive.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7477970 TI - Plasma extravasation in the skin and pelvic organs evoked by antidromic stimulation of the lumbosacral dorsal roots of the rat. AB - Electrical stimulation of the distal stump of cut peripheral nerves is a commonly accepted way to evoke neurogenic inflammation. Nevertheless, the modulatory effect of biogenic amines and vasoactive peptides released from efferent fibres can be excluded only if the dorsal roots are stimulated. The present study was focussed to investigate plasma extravasation in the appropriate skin and mucosal areas as well as in the genito-urinary organs in response to antidromic stimulation of the lumbar and sacral dorsal roots of the rat. Plasma extravasation was detected by quantitative measurement of the accumulated Evans Blue tracer in tissue pieces. Two unilateral posterior roots were stimulated simultaneously (20 V, 0.5 ms, 5 Hz, 5 min) in each anaesthetized rat. Intensive blueing response occurred in the following tissues: plantar glabrous skin, L4-L5 (L6); dorsum of the hindpaw and ankle joint, L2-L4; ventral surface of the thigh, L2-L4 (L1); abdominal skin, L1-L4; caudal nipples, L1-L2; root of the tail, S1 orifice of the vagina, S1 (L6); vagina, L2-L3, L5-S1; cervix and corpus uteri, L2 L3, L5-S1; lower two-thirds of the uterine horns, L1-L3; urinary bladder, L1-L3, L6-S1; rectum, L5-S1; scrotum (dorsal surface and lower pole), L6-S1; scrotum (ventral surface), L3-L5. No significant dye accumulation was observed in the muscles, testicles, vas deferens and prostate. Plasma extravasation caused by antidromic activation of the dorsal roots was absent or highly reduced after systemic capsaicin pretreatment of the rats. Neurogenic inflammation evoked by antidromic stimulation of the dorsal roots makes this method suitable for mapping the organs where capsaicin-sensitive sensory nerve endings exert their "efferent functions". This first functional description of segmental innervation of capsaicin-sensitive afferent fibres is in agreement with retrograde tracing studies and immunohistochemical localization of substance P in the dorsal root ganglia and peripheral tissues. PMID- 7477971 TI - Neurologic complications of lumbar epidural anesthesia and analgesia. AB - We reviewed the clinical features of 12 patients with neurologic complications following lumbar epidural anesthesia or analgesia. Eleven patients experienced lumbosacral radiculopathy or polyradiculopathy and, of these, 10 received epidural anesthesia or analgesia and one received subarachnoid injection of medication after intended epidural anesthesia. One patient suffered a moderately severe thoracic myelopathy in the setting of unintended spinal anesthesia. The two patients with more severe polyradiculopathy had severe lumbar spinal stenosis on MRI. The other patients experienced mild to moderate neurologic deficits most often involving the L-2 root, and MRIs, when performed, were unremarkable. EMG on three patients helped to localize the lesions to the lumbosacral roots and to quantify the extent of axonal loss. Ten patients were ambulatory upon discharge from the hospital and had good neurologic outcome. One patient with severe polyradiculopathy did not improve after 4 years and had severe motor axonal loss based upon electrodiagnostic studies. The patient with a thoracic myelopathy was ambulatory 4 months after onset. Although generally a safe procedure with low frequency of complications, lumbar epidural anesthesia or analgesia occasionally causes neurologic sequelae such as radiculopathy or myelopathy. Neurologic complications may be more severe in the presence of spinal stenosis or after inadvertent subarachnoid injection of anesthetic or analgesic agent. PMID- 7477972 TI - Meningeal biopsy in intracranial hypotension: meningeal enhancement on MRI. AB - Intracranial hypotension is a cause of diffuse enhancement of the pachymeninx with gadolinium, which often is associated with subdural fluid collections. We reviewed the results of meningeal biopsy in six patients with intracranial hypotension and diffuse pachymeningeal enhancement to correlate the MRI findings with histopathologic observations and to explain the abnormalities seen on MRI. Grossly, the dura mater was unremarkable in all patients, as were the leptomeninges, except for one patient with prolonged (18 months) intracranial hypotension in whom the arachnoid was thickened and opaque. Microscopically, the dura mater was entirely normal on its epidural aspect; however, a fairly thin zone of fibroblasts and thin-walled small blood vessels in an amorphous matrix was noted on the subdural aspect. In the patient with longstanding symptoms, diffuse benign arachnoidal cell proliferation was also noted, probably a reaction triggered by longstanding changes in the subdural area, as noted in the five other patients. There was no evidence of inflammation, infection, or metastatic neoplasia. These findings suggest that in intracranial hypotension, the dural meningeal abnormalities probably represent reactive secondary phenomena, likely related to hydrostatic changes in the CSF, and not a primary meningeal process. PMID- 7477973 TI - Changes in the amount of diseased white matter over time in patients with relapsing-remitting multiple sclerosis. AB - MRI is a sensitive technique for assessing disease activity in MS. Diseased white matter (WM) can be identified on T2-weighted images, and active disease is reflected by abnormalities in the blood-brain barrier (BBB) shown on T1-weighted images after administration of paramagnetic contrast agents. Active disease may be demonstrated by contrast-enhanced MRI in patients with early, mild relapsing remitting (RR) MS even during periods of clinical stability, which indicates that MS is an active process even during the early phase of the illness. To examine the amount of abnormal WM at frequent intervals over time, we studied seven mildly affected RRMS patients, all of whom had frequent contrast-enhancing lesions. These RRMS patients were imaged monthly for 26 to 36 months at 1.5 tesla; the area of abnormal increased WM signal was calculated by image processing software that utilizes both the T2- and T1-weighted images. All patients showed fluctuations over time in amount of abnormal WM signal, which reflected factors such as the amount of BBB breakdown (measured by number or area of enhancing lesions) and measurement error. All seven RRMS patients, however, showed an overall increase in abnormal WM. Because of the fluctuations between individual measurements, the increase was most accurately reflected when the mean of the first 6 months' measurements was compared with the mean of the final 6 months' measurements, or when a linear regression model was applied.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7477974 TI - Involvement of the healthy hemisphere in recovery from aphasia and motor deficit in patients with cortical ischemic infarction: a transcranial Doppler study. AB - Activation of areas of the health hemisphere seems to play a role in functional recovery from stroke. We studied cerebral blood flow changes during motor and mental activity in patients with cortical ischemic lesions. We simultaneously measured blood flow velocity in the two middle cerebral arteries of 45 patients with single cortical ischemic lesions and good functional recovery and of 16 healthy controls by means of bilateral transcranial Doppler ultrasonography during a 2-minute sequential thumb-to-finger opposition task, alternately performed with the right and left hands, and during a 1-minute word-fluency task. Twenty-five patients had left cortical lesions, 12 with previous motor deficit alone and 13 with associated motor deficit and Broca's aphasia. Twenty patients had right cortical lesions with previous motor deficit. With respect to baseline values, the increase of flow velocity in the middle cerebral artery contralateral to the hand performing the motor task was comparable in controls and patients, regardless of the side of the lesion and the hand (normal or recovered) involved in the task. During movement of the recovered hand, the increase of flow velocity in the ipsilateral middle cerebral artery was significantly greater (p < 0.001, two-way ANOVA) than the increase during movement of the normal hand in both controls and patients. During performance of the word-fluency task, the increase of flow velocity in the left middle cerebral artery was comparable in controls and patients.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7477975 TI - Functional MRI during word generation, using conventional equipment: a potential tool for language localization in the clinical environment. AB - OBJECTIVE: To test the accuracy of bilateral language mapping using a standard clinical magnetic resonance (MR) imaging device during word generation. DESIGN: A study of normal volunteers. SETTING: Volunteers from the Washington, DC, area. PARTICIPANTS: Nine normal, right-handed, native English speakers (four women, five men, mean age 31 years). INTERVENTIONS: During four MR acquisition periods, subjects would alternately rest and silently generate words. Sagittal MR images covered the middle and inferior frontal gyri, insulae, and part of the temporal and parietal lobes bilaterally. MAIN OUTCOME MEASURES: (1) Anatomic maps of task related signal changes obtained by comparing, in each voxel, the signal during word generation and rest periods, and (2) analysis of the time course of the signal. RESULTS: Maximum responses were in the left hemisphere, mainly in the frontal lobe (Broca's area, premotor cortex, and dorsolateral prefrontal cortex) but also in posterior regions such as Wernicke's area. In agreement with previous studies, some degree of task-related changes was present in a subset of the corresponding regions in the right hemisphere. CONCLUSION: Despite certain limitations, it is possible, using widely available MR equipment, to obtain results consistent with previous studies. The technique may have important implications for assessment of cognitive functions in patients with neurologic disorders in a clinical environment. PMID- 7477976 TI - Multifocal demyelinating motor neuropathy: pathologic evidence of 'inflammatory demyelinating polyradiculoneuropathy'. AB - We report a case of multifocal demyelinating motor neuropathy in a patient with a 5-year history of progressive, asymmetric, predominantly motor weakness characterized by multifocal progression, multifocal conduction block, and lack of response to steroid therapy. Neuropathologic findings at autopsy showed an "inflammatory demyelinating polyradiculoneuropathy" in the motor cranial nerves and motor roots of peripheral nerves, an extensive deposition of IgG and focal accumulations of IgM in the peripheral nerve motor roots, and loss of motor neurons. These findings clearly document an inflammatory demyelinating polyradiculoneuropathy in multifocal demyelinating motor neuropathy, suggesting a close relation with chronic inflammatory demyelinating polyneuropathy. PMID- 7477977 TI - Chronic symmetric symptomatic polyneuropathy in the elderly: a field screening investigation in two Italian regions. I. Prevalence and general characteristics of the sample. Italian General Practitioner Study Group (IGPSG). AB - The prevalence and general characteristics of chronic symmetric symptomatic polyneuropathy were assessed in two elderly populations living in Varese, northern Italy, and San Giovanni Rotondo, southern Italy. We interviewed 4,191 subjects (3,027 in Varese and 1,164 in San Giovanni Rotondo), 55 years and older, seen in office consultations by 27 general practitioners. A neurologist examined 734 patients who had two or more symptoms of polyneuropathy. A diagnosis of possible polyneuropathy (screening neuropathic symptoms and one of the following findings: bilateral impairment of strength; bilateral impairment of sensation; bilateral impairment of deep tendon reflexes) was made in 213 patients (7.0%) in Varese and 94 (8.1%) in San Giovanni Rotondo. Probable polyneuropathy (screening symptoms and at least two of the physical findings) was present in 111 Varese patients (3.7%) and 40 San Giovanni Rotondo patients (3.4%). The age- and sex adjusted prevalence rate of probable polyneuropathy was 3.6 per 100 in Varese and 3.3 per 100 in San Giovanni Rotondo. The disease was more prevalent in women in Varese and in men in San Giovanni Rotondo and was significantly correlated with age in Varese. Diabetes was found in association with probable polyneuropathy in 43.7% of patients. Muscle cramps and distal paresthesia were the main symptoms. In general, polyneuropathy was mild to moderate, impairment of deep tendon reflexes and sensation being the most common findings. PMID- 7477978 TI - 'Complicated migraine-like episodes' in children following cranial irradiation and chemotherapy. AB - Neurologic sequelae may occur months to years after cranial irradiation. The site of primary damage is probably the vascular endothelium. Over a 2.8-year period, four children with brain tumors, a mean of 11 years of age at diagnosis (range, 6.5 to 15.5 years), had new onset of severe intermittent unilateral headaches associated with nausea, episodic visual loss, hemiparesis, aphasia, or hemisensory loss. The headaches lasted 2 to 24 hours. All patients had previously received whole-brain (2,400 to 3,600 cGy) and additional local boost (1,800 to 3,100 cGy) cranial irradiation, as well as cisplatin-, lomustine-, and vincristine-containing chemotherapy regimens. Symptoms began 1.2 to 2.8 years after the diagnosis, when all had stable disease and were off treatment. MRI studies were unchanged, and CSF cytology, EEGs, echocardiograms, and magnetic resonance angiograms were normal in all. Cerebral angiograms, performed in three children, were normal but led to severe headaches and neurologic deficits (hemiparesis in one and visual loss in two) that resolved after 24 to 48 hours. Response to antimigraine and antiplatelet medications was variable. We conclude that (1) "complicated migraine-like episodes" may occur in children after cranial irradiation and chemotherapy as a sequela of therapy; (2) these headaches may not be the harbinger of impending strokes, severe intracranial vasculitis, or tumor recurrence; and (3) while cerebral angiography may be useful in differential diagnosis, it may cause transient worsening of symptoms. PMID- 7477979 TI - Interictal metabolism and blood flow are uncoupled in temporal lobe cortex of patients with complex partial epilepsy. AB - We used positron emission tomography (PET) with 18F-2-deoxyglucose (FDG) and 15O water in 20 patients with complex partial seizures to compare glucose metabolism and blood flow in temporal lobe epileptic foci identified by ictal scalp sphenoidal video-EEG telemetry. Glucose metabolism was measured 20 minutes after blood flow without moving the patient from the scanner. We also studied 11 patients with 99mTc-HMPAO single-photon emission computed tomography (SPECT). Both local cerebral metabolic rate of glucose (LCMRGlc) and regional cerebral blood flow (rCBF) were significantly decreased in temporal cortex ipsilateral to the EEG focus. However, LCMRGlc was reduced by 11.2% in inferior lateral and 11.1% in inferior mesial temporal cortex and rCBF by only 3.2% and 6.1%. The ratio of LCMRGlc to rCBF was significantly reduced in inferior lateral temporal cortex ipsilateral to the ictal focus (p < 0.009). Moreover, using standardized criteria, blinded raters found that 16 of 20 patients had focal FDG-PET hypometabolism, all in the epileptogenic region; 10 of 20 had focal 15O water PET hypoperfusion, but it was falsely lateralized in two of these 10; and five of 11 had focal 99mTc-HMPAO SPECT hypoperfusion, but it was falsely lateralized in two of these five. Our data suggest that interictal glucose metabolism and blood flow may be uncoupled in epileptogenic cortex. PMID- 7477980 TI - Cutaneous innervation in sensory neuropathies: evaluation by skin biopsy. AB - OBJECTIVE: To use punch skin biopsies to evaluate the loss of intra-epidermal nerve fibers in sensory neuropathies. BACKGROUND: Previous assessments of epidermal nerve fibers have been constrained by relatively insensitive staining techniques and variability in quantification. METHODS: Punch skin biopsies were performed on the heel and leg of HIV-seronegative controls, HIV-seropositive individuals without neuropathy, and patients with sensory neuropathies, including HIV-seronegative and HIV-positive individuals. After formalin fixation, 50 microns free-floating sections were stained with a monoclonal antibody to neuron specific ubiquitin hydrolase, PGP9.5. The number of intraepidermal fibers/mm in at least three sections from each patient was counted by one observer blinded to site and clinical status. RESULTS: Dermal and epidermal nerve fibers were readily identified and quantified. The immunostaining technique reliably demonstrated a dermal plexus of myelinated and unmyelinated fibers parallel to the surface of the skin. In the epidermis, unmyelinated fibers ascended vertically between the keratinocytes to reach the stratum corneum. The number of intra-epidermal fibers/mm in the distal leg (mean +/- SEM) was 17.84 +/- 3.03 in seven HIV seronegative controls. Epidermal fiber number was significantly reduced (p = 0.01) in five HIV-infected patients with sensory neuropathies associated with didanosine or zalcitabine therapy (1.07 +/- 0.40) and in eight HIV-seronegative patients with sensory neuropathies (3.1 +/- 3.1). Four of five neurologically normal HIV-seropositive subjects had reduced numbers of epidermal fibers, suggesting a subclinical neuropathy. Serial biopsies in one individual demonstrated the evolution of degenerating epidermal fibers after development of zalcitabine-induced sensory neuropathy. CONCLUSION: Skin biopsies stained with the sensitive panaxonal marker anti-PGP9.5 demonstrated significant reduction in intraepidermal fibers in sensory neuropathies. This simple and repeatable technique is a reliable method for quantitation of small cutaneous sensory fibers. In addition, skin biopsies may be useful in assessing the course and spatial distribution of involvement in peripheral nerve disease. PMID- 7477981 TI - Levodopa-induced dyskinesias are improved by fluoxetine. AB - We evaluated the severity of motor disability and dyskinesias in seven levodopa responsive patients with Parkinson's disease after an acute challenge with the mixed dopamine agonist, apomorphine, before and after the administration of fluoxetine (20 mg twice per day) for 11 +/- 1 days. After fluoxetine treatment, there was a significant 47% improvement (p < 0.05) of apomorphine-induced dyskinesias without modification of parkinsonian motor disability. The dyskinesias were reduced predominantly in the lower limbs during the onset and disappearance of dystonic dyskinesias (onset- and end-of-dose dyskinesias) and in the upper limbs during choreic mid-dose dyskinesias. The results suggest that increased brain serotoninergic transmission with fluoxetine may reduce levodopa- or dopamine agonist-induced dyskinesias without aggravating parkinsonian motor disability. PMID- 7477982 TI - The T-cell receptor beta locus and susceptibility to multiple sclerosis. AB - Assessments of genetically determined variations in the T-cell antigen receptor in multiple sclerosis (MS) have yielded conflicting results. We used three restriction fragment length polymorphisms (RFLPs) and a polymorphic microsatellite repeat as markers for the T-cell receptor (TCR) beta locus (7q32 35) in multiplex MS families. Affected sibling-pair analysis of the RFLP data failed to show evidence for linkage (127 families) whereas analysis of the microsatellite data (86 families) provided weak evidence for linkage with a maximum lod score of 0.98 (p < 0.05). We repeated the analysis in those families (n = 53) in which the affected sibling pairs were concordant for the HLA haplotype DR15/DQ6. This altered the proportion of affected siblings sharing 0, 1, 2 RFLP haplotypes from 0.24, 0.50, and 0.26 (p = NS) before stratification to 0.16, 0.41, and 0.43 (p < 0.05) in the DR15/DQ6 positive pairs alone; for the microsatellite data, sharing altered from 0.16, 0.50, and 0.34 (p < 0.05) in all pairs to 0.07, 0.49, and 0.44 (p < 0.01) in the DR15/DQ6 concordant siblings. PMID- 7477983 TI - New connexin32 mutations associated with X-linked Charcot-Marie-Tooth disease. AB - Analysis of the connexin32 gene in patients with X-linked Charcot-Marie-Tooth disease shows mutations distributed throughout the molecule, with all domains affected except the fourth transmembrane domain and the distal carboxy terminus. Sequence analysis of DNA from 19 unrelated patients detected six novel mutations and three previously reported mutations. Identification of additional mutations extends the distribution of connexin32 mutations in X-linked Charcot-Marie-Tooth disease and shows that specific mutations recur in additional families. PMID- 7477985 TI - Immunization with the paraneoplastic encephalomyelitis antigen HuD does not cause neurologic disease in mice. AB - Paraneoplastic encephalomyelitis/paraneoplastic sensory neuronopathy (PEM/PSN) associated with small cell lung cancer is characterized by high serum and CSF titers of anti-neuronal (anti-Hu) antibodies and by intrathecal synthesis of anti Hu IgG. A pathologic role for the anti-Hu antibodies in PEM/PSN is further suggested by reported intraneuronal accumulation of the antibodies in the nervous system of PEM/PSN patients at autopsy. We immunized SJL/J mice, Lewis rats, and Hartley guinea pigs with purified recombinant HuD fusion protein. In spite of high-titer anti-HuD antibodies, neurologic and pathologic examination of the animals was normal. Apparent uptake of purified IgG by neurons in the brain proved to be artifactual. PMID- 7477984 TI - A prospective evaluation of cognitive decline in early Huntington's disease: functional and radiographic correlates. AB - We examined prospectively the relationship between progressive disability in Huntington's disease (HD) and concomitant alterations in neuropsychological functioning and brain imaging indices in a cohort of 60 patients who were enrolled and followed for 30 to 42 months in a controlled clinical trial. Standardized measures of functional capacity and neuropsychological performance were collected, and CT was performed, at regular intervals every 6 to 12 months. Psychomotor skills showed the most significant and consistent decline among the cognitive functions assessed. Memory disturbances were already present at the time of enrollment, but memory did not deteriorate until patients reached advanced stages. Other cognitive operations, such as visual construction and semantic knowledge, manifested small and variable changes over time. CT indices of striatal atrophy correlated only with changes in psychomotor function, while the CT index of frontal atrophy weakly predicted memory and semantic knowledge scores at 42 months. These results confirmed earlier cross-sectional findings and extended our knowledge of the evolution of cognitive dysfunction in HD. PMID- 7477986 TI - Marked increase in mitochondrial DNA deletion levels in the cerebral cortex of Huntington's disease patients. AB - To determine if somatic mtDNA mutations might contribute to the neurodegeneration observed in Huntington's disease (HD), we quantitated the amount of the common mitochondrial 4977 nucleotide pair deletion (mtDNA4977) in cortex and putamen of HD patients and age-matched controls by the serial dilution-polymerase chain reaction method. Cortical deletion levels were analyzed in the temporal, frontal, and occipital lobes. HD temporal lobes had an 11-fold greater mean mtDNA4977 deletion level than age-matched controls, and HD frontal lobes had fivefold greater levels. HD occipital lobe and putamen deletion levels were comparable with control levels. These results support the hypothesis that HD is associated with elevated cortical mtDNA damage. PMID- 7477987 TI - Involvement of glycine and GABAA receptors in the pathogenesis of spinal myoclonus: in vitro studies in the isolated neonatal rodent spinal cord. AB - Studies of pathogenetic mechanisms of myoclonus and spinal spasticity have established a strong association between deficient inhibitory glycinergic transmission and pathologic rigidity and tremor. Consistent with known cases in the clinical literature, electrophysiologic data from animal models of myoclonus implicate dysfunction of segmental spinal cord circuitry. The present study sought to further explore pathogenetic mechanisms at the circuit level. In vitro preparations of isolated spinal cord from neonatal rodents allowed for stable recordings of individual cells as well as populations of motoneurons. Blockade of glycine receptors enhanced 5- to 15-Hz sinusoidal oscillations that were synchronous in entire populations of motoneurons as well as along multiple segments of the spinal cord. Oscillations at motoneurons were mediated largely by non-NMDA excitatory synaptic inputs. Blockade of GABAA receptors, and not GABAB receptors, abolished sinusoidal oscillations, suggesting a critical role for GABAA receptors in the premotoneuronal circuitry responsible for generation or transmission of the sinusoidal oscillations. These data offer new insights into possible pathogenetic mechanisms of spinal myoclonus and may help guide future research leading to specific therapies for hyperkinetic movement disorders of spinal origin. PMID- 7477989 TI - Brainstem tumors in patients with neurofibromatosis type 1: a distinct clinical entity. AB - The natural history and the clinical and neuroimaging features of brainstem tumors in neurofibromatosis type 1 (NF1) are poorly understood. Magnetic resonance imaging (MRI) has been useful in NF1 in detecting intracranial abnormalities, especially of the brainstem. Brainstem tumors in NF1 have been confused clinically with non-NF1 brainstem tumors and radiographically with the increased T2 signal abnormalities, also known as "unidentified bright objects" (UBOs), which are common in NF1 and often located in the brainstem. This study, which evaluated 17 NF1 patients with brainstem tumors, is the largest series to date. Fifteen of 17 patients (88%) had neurologic signs and symptoms referable to brainstem dysfunction, including dysarthria, cranial neuropathies, and gross motor incoordination. Tumors were located primarily in the medulla in 14 of 17 NF1 patients (82%), in contrast to the pontine tumor location in the non-NF1 population. Seven NF1 patients (41%) required shunt placement for hydrocephalus at initial diagnosis, more frequent than in non-NF1 brainstem tumor patients. Six of 17 patients (35%) had evidence of radiographic tumor progression, but only three of them (18%) had correlative clinical progression. Two patients with progressive symptoms had partial surgical resection, and pathology revealed either fibrillary or anaplastic astrocytomas. Three patients were treated with radiation therapy, chemotherapy, or both, with two deaths. With a median follow up of 52 months, 15 of 17 patients remain alive; 14 of them did not require adjuvant therapy. In our series, we describe NF1 brainstem tumors as a distinct clinical entity, much less aggressive than non-NF1 pontine tumors but more symptomatic than brainstem UBOs in NF1. PMID- 7477988 TI - Intrathecal baclofen therapy in stiff-man syndrome: a double-blind, placebo controlled trial. AB - We performed a double-blind, placebo-controlled trial of intrathecal baclofen (ITB) in stiff-man syndrome. Three patients, unresponsive to current therapy, received 50 micrograms of ITB or placebo on sequential days. Following ITB, all patients demonstrated improvement in reflex EMG activity. The mean reduction in total EMG activity (from all muscles) following stimulation of the medial plantar nerve (cutaneous flexor reflex) was 72% following 50 micrograms of ITB compared with 18% following placebo (ANOVA: significance of F, p < 0.0001). The mean latency to onset of the response was also significantly prolonged for all muscles following ITB (ANOVA: significance of F, p < 0.05). Although reflex EMG activity was reduced in all patients, clinical improvement was evident in only one patient, who differed from the others studied by a longer duration of disease, greater severity of stiffness, less fear of falling, and greater electrophysiologic improvement. PMID- 7477990 TI - Erythrocyte thiolmethyltransferase: another failed marker for Alzheimer's and Parkinson's diseases. AB - There are reports that patients with Parkinson's disease (PD) and Alzheimer's disease (AD) have reduced levels of thiolmethyltransferase (TMT) in erythrocyte membranes. TMT methylates thiols and thiocarbamates, thereby reducing their toxicity. We examined TMT levels in erythrocytes from patients with PD and AD and from age-matched controls. Specific activities of TMT were 564 +/- 199 U/mg protein in PD (n = 32), 513 +/- 118 in AD (n = 13), and 565 +/- 183 in controls (n = 35). There was no difference between any of the groups (p = 0.64). We failed to confirm TMT as a marker for neurodegenerative diseases or for this metabolic defect predisposing to susceptibility to neurotoxins. PMID- 7477991 TI - Similar potency of carbamazepine, oxcarbazepine, and lamotrigine in inhibiting the release of glutamate and other neurotransmitters. AB - We compared the effects of the antiepileptic drugs carbamazepine, oxcarbazepine, and lamotrigine on the release from rat brain slices of endogenous glutamate, [3H]-GABA, and [3H]-dopamine, elicited by the Na+ channel opener, veratrine, and of the same transmitters as well as [3H]-noradrenaline, [3H]-5-hydroxytryptamine, and [3H]-acetylcholine, elicited by electrical stimulation. The three antiepileptic drugs inhibited veratrine-induced release of endogenous glutamate, [3H]-GABA, and [3H]-dopamine, with IC50 values between 23 and 150 microM, in or near the concentration range in which they interact with Na+ channels, and there was little difference between the compounds. They were five to seven times less potent in inhibiting electrically as compared with veratrine-stimulated release of [3H]-GABA and [3H]-dopamine; similarly, carbamazepine and tetrodotoxin were more potent in inhibiting veratrine-induced as compared with electrically induced release of endogenous glutamate. Carbamazepine, oxcarbazepine, and lamotrigine also inhibited electrically stimulated release of [3H]-5-hydroxytryptamine (IC50 values, 150 to 250 microM) and [3H]-acetylcholine (IC50 values, 50 to 150 microM); [3H]-noradrenaline release was affected to a lesser degree. The active concentration ranges of these drugs with respect to inhibition of veratrine stimulated neurotransmitter release matched the therapeutic plasma and brain concentrations. It is uncertain whether these effects are relevant in vivo at anticonvulsant doses, because the drugs are markedly less potent in inhibiting the more physiologic release elicited by electrical stimulation. Therefore, the hypothesis that inhibition of glutamate release is the mechanism of anticonvulsant action of lamotrigine (or carbamazepine and oxcarbazepine) is doubtful. Other consequences of Na+ channel blockade may have an important role. PMID- 7477992 TI - Soluble E-selectin in the serum and cerebrospinal fluid of patients with multiple sclerosis and human T-lymphotropic virus type 1-associated myelopathy. AB - To evaluate the activation of endothelial cells of the brain and the spinal cord, we investigated the presence of soluble endothelial leukocyte adhesion molecule-1 (sE-selectin) in the serum and cerebrospinal fluid (CSF) of patients with multiple sclerosis (MS) and those with human T-lymphotropic virus type 1 associated myelopathy (HAM). There were significantly higher levels of sE selectin found in the serum of patients with relapsing-remitting MS during an exacerbation (p < 0.001) and those with chronic progressive MS (p < 0.01) compared with controls. Serum levels of sE-selectin in patients with HAM did not differ significantly from serum levels in controls or non-HAM carriers. We also found sE-selectin in the CSF of eight patients during an exacerbation of relapsing-remitting MS. These results suggest that an active immune reaction involving E-selectin production that is indicative of endothelial cell damage occurs in the CNS of patients during an exacerbation of relapsing-remitting MS. Thus, sE-selectin may be useful in monitoring disease activity in patients with relapsing-remitting MS. PMID- 7477993 TI - Diversity of T-cell receptor V alpha, V beta, and CDR3 expression by myelin basic protein-specific human T-cell clones. AB - We sequenced the cDNAs of alpha and beta T-cell receptors (TCRs), including V, J, and CDR3 regions, expressed by 54 myelin basic protein (MBP)-specific T-cell clones generated from the peripheral blood of 15 multiple sclerosis (MS) patients and three normal controls. Thirteen V alpha gene segments, 18 V beta gene segments, 23 CDR3 alpha sequences, and 30 CDR3 beta sequences were represented among these clones. Some CDR3 motifs were common to several clones that shared epitope specificity, while other motifs were common to clones with diverse epitope specificities. The extensive heterogeneity of TCR gene expression in the human immune response to MBP indicates that therapeutic strategies aimed at blocking a limited number of TCRs are unlikely to fully suppress the T-cell response to MBP in vivo. PMID- 7477995 TI - Anticonvulsant effect of fluoxetine in humans. AB - We report an unblinded, open-label, add-on trial of fluoxetine, a selective serotonin reuptake inhibitor, in 17 patients with complex partial seizures with and without secondary generalization (mean follow-up duration, 14 +/- 1.1 months). Six patients showed complete disappearance of their daily seizures; in the others the seizure frequency was lowered by 30%. No patient reported side effects. PMID- 7477994 TI - Low-dose, short-term oral prednisone in the treatment of carpal tunnel syndrome. AB - We evaluated the effectiveness of low-dose, short-term oral prednisone in ameliorating the pain and other symptoms of carpal tunnel syndrome (CTS) in a randomized, double-blind, placebo-controlled study of patients with mild to moderate CTS. Prednisone, in doses of 20 mg daily for the first week and 10 mg daily for the second week, resulted in significant improvement in global symptom scores. The effect was rapid, but gradually waned over 8 weeks of observation. This approach may provide a treatment alternative in the short-term, conservative management of CTS. PMID- 7477996 TI - Tapping the head activates the vestibular system: a new use for the clinical reflex hammer. AB - We investigated the use of skull taps with a modified clinical reflex hammer as a method of vestibular activation. Using recently described EMG techniques to measure vestibulocollic reflexes in response to clicks, we were able to show analogous short-latency potentials to taps. The earliest responses were invariably absent on the side of a previous vestibular nerve section but were preserved in profound sensorineural or conductive hearing loss. We propose that the taps activated the vestibular apparatus directly by a bone-conducted vibration wave. PMID- 7477997 TI - Middle cerebral artery dissection: a clinicopathologic study. AB - We present a case report of a 17-year-old young man who developed fatal dissection of the middle cerebral artery after what appeared to be trivial trauma. The dissection was not evident on cerebral arteriogram but was identified at autopsy. Arterial dissection should be considered in the differential diagnosis of supraclinoid occlusion of the internal carotid artery seen by cerebral angiography. PMID- 7477998 TI - MRI of Creutzfeldt-Jakob disease: asymmetric high signal intensity of the basal ganglia. AB - We report a pathologically proven case of Creutzfeldt-Jakob disease (CJD) with striking asymmetry in abnormal high signal intensity of the basal ganglia on long repetition-time MRI. Symmetric or asymmetric high signal intensity of the basal ganglia, in the appropriate clinical setting, may be a specific finding of CJD. PMID- 7477999 TI - Does homozygosity advance the onset of dentatorubral-pallidoluysian atrophy? AB - We confirmed a homozygous type of relatively small expansion of CAG triplet repeats (57 repeats) in the short arm of chromosome 12 in a male patient with dentatorubral-pallidoluysian atrophy (DRPLA) by polymerase chain reaction. He showed early onset (age, 17 years) of DRPLA. There was good correlation of the age of onset with the number of triplet repeats. The homozygous state of the expansion of the triplet repeats was responsible for the early onset and severity of his DRPLA. PMID- 7478000 TI - Guidelines on the detection of paraneoplastic anti-neuronal-specific antibodies: report from the Workshop to the Fourth Meeting of the International Society of Neuro-Immunology on paraneoplastic neurological disease, held October 22-23, 1994, in Rotterdam, The Netherlands. PMID- 7478001 TI - Subtle extrapyramidal signs and incident dementia: a follow-up analysis. PMID- 7478002 TI - EEG during MR imaging: differentiation of movement artifact from paroxysmal cortical activity. PMID- 7478003 TI - Lumbosacral plexopathy due to benign uterine leiomyoma. PMID- 7478004 TI - Neuronal heterotopia. PMID- 7478005 TI - Practice parameters. PMID- 7478006 TI - Idiopathic recurring stupor. PMID- 7478007 TI - Hippocampal sclerosis. PMID- 7478008 TI - Hemicrania continua. PMID- 7478009 TI - Brain herniation and mannitol. PMID- 7478010 TI - Post-traumatic movement disorders. PMID- 7478011 TI - Post-traumatic movement disorders. PMID- 7478012 TI - Felbamate and tremor. PMID- 7478013 TI - Demonstration of the venous systems with gadolinium-enhanced three-dimensional phase-contrast MR venography. AB - The purpose of this study was to evaluate the usefulness and advantages of gadolinium-enhanced three-dimensional phase-contrast MR venography for demonstrating the venous systems. The three-dimensional phase-contrast MR venography was performed with a velocity encoding gradient settings from 5 to 20 cm/s on 22 normal subjects. In 8 of normal subjects, gadolinium-enhanced phase contrast MR venography was performed. 22 subjects (100%) had detectable flow in the sphenoparietal sinus, transverse sinus, basal vein, and internal cerebral vein. With a VENC setting at 10 cm/s, venous system was visualized selectively and clearly. Detection ratio in inferior petrosal sinus, superior petrosal sinus, and superior ophthalmic vein increased from 0% to 25%, from 28.6% to 62.5%, and from 28.6% to 37.5%, respectively, after administration of gadopentate dimeglumine. In conclusion, gadolinium-enhanced three-dimensional phase-contrast MR venography was useful for demonstrating the venous systems. PMID- 7478015 TI - Intranasal (transethmoidal) encephalomeningocele. Case report. AB - Intranasal encephaloceles are rarely encountered in pediatric neurosurgery. The symptoms and clinical features may mimic those of nasal polyp. It is important to know the type of basal encephalomeningocele for appropriate surgical intervention. Computed tomographic examination is helpful for differential diagnosis of the encephalocele sac and localization of the cranial bone defect. PMID- 7478014 TI - Delayed post-traumatic epidural hematoma. A review. AB - Post-traumatic acute epidural hematoma (EDH) is generally visible on the CT scan done immediately after admission: occasionally, it only comes to light at a later scan and is then termed delayed (DEDH). Since the introduction of CT, the frequency of this occurrence has gone up from 6-13% to 30%. The mechanisms responsible for the delayed appearance of the epidural hematoma a "tamponade" effect are usually increased endocranial pressure and post-traumatic arterial hypotension as well as, in a limited number of cases, coagulopathy, CSF drainage, and arterio-venous shunt. The authors report 5 of their own cases and 45 published cases and discuss the characteristics of this particular form of hematoma and its outcome. PMID- 7478017 TI - Giant fusiform aneurysm of the vertebro-basilar artery presenting with stroke. AB - The authors describe a case of giant fusiform aneurysm of the basilar artery presenting with ischemic symptoms. Angiography and CT revealed vertebro-basilar fusiform aneurysmal dilatation. Fusiform vertebro-basilar aneurysm is associated with various complications particularly brain stem infarction. Similar lesions in the literature are reviewed and the relationship between this clinical entity and cerebral ischemia, particularly brain-stem infarction are discussed. PMID- 7478016 TI - Thoracic and lumbar meningocele in neurofibromatosis type 1. Report of two cases and review of the literature. AB - We report two cases of ventrolateral thoracic and lumbar meningoceles associated with neurofibromatosis type 1, and present a review of the literature. Thoracic and lumbar meningoceles are rare lesions which are associated with neurofibromatosis in 60% to 85% of cases. Large meningoceles may cause pain, neurological signs, or respiratory complaints when located intrathoracically. Surgery is indicated in such cases, as well as when small meningoceles enlarge with time. The etiology of meningoceles in neurofibromatosis remains controversial. The authors postulate that ventrolateral lesions of the thoracic and lumbar spine are secondary to congenital mesodermal dysplasia and hypoplastic bone changes. MRI is the diagnostic method of choice, since it most adequately visualizes the pathological anatomy of the spinal cord, meninges, and adjacent structures in every sectional plane. CT scans are particularly useful in assessing the spinal bony changes. PMID- 7478018 TI - Solitary plasmacytoma of the skull. A case report. AB - A case of solitary plasmacytoma of the skull is presented. The tumor was radically removed. There was no systemic involvement and the patient was not submitted to radiotherapy. Lytic lesions of the skull are discussed. PMID- 7478019 TI - Primary leptomeningeal melanoma simulating a meningioma. AB - A surgically confirmed primary leptomeningeal malignant melanoma (PLMM) discovered at the parietal region is reported in a 72-year-old male. He developed progressive right hemiparesis and speech disorders caused by a parietal large mass that simulated a growing meningioma. A well-defined, dark-black tumor was removed completely and was histopathologically diagnosed as a malignant melanoma. No melanomas were detected by systemic clinical and radiological examination, including dermatological and ophthalmologic examinations. Follow-up examination 18 months postoperatively showed no evidence of recurrence of the tumor. The patient gradually became bedridden probably because of decreased general activity possibly due to brain atrophy and died of cardiac failure without any evidence of recurrence. Because of the absence of other systemic localizations, we consider this melanoma as primary. A favorable outcome was obtained by surgical treatment alone. PMID- 7478020 TI - Balloon angioplasty immediately after surgical clipping for symptomatic vasospasm on admission. Report of four cases. AB - In spite of recent advancements in the management of ruptured aneurysm, there are still controversies as to how to threat the patients in serious conditions, one of these concerning whether to operate patients admitted with symptomatic vasospasm (vasospasm on admission) and when. While early surgery may protect patient from further rupturing, it may also increase risk of worsening by inducing much more vasospasm. Four cases of severe vasospasm associated with aneurysmal subarachnoid hemorrhage (SAH) were reported. All of these cases showed symptomatic and angiographical vasospasm on their admission 3-11 days after initial attack of SAH. All cases had a preoperative clinical grading using Hunt & Kosnik of IV, and were treated by early surgery with clipping followed by percutaneous balloon angioplasty (PTA) immediately after clipping. Two cases returned to their previous occupations, while one case remained bed ridden and one died in spite of therapy. Early surgery in this series actually prevented further rupturing, but it is not clear whether PTA immediately after clipping may prevent further deterioration by progression of vasospasm. For selected cases especially those with reversible ischemia, this combined treatment may offer more favorable results than late surgery with conservative medical care. Further investigation is necessary to support the validity of this therapy. PMID- 7478021 TI - Pre- and intraoperative methods of controlling cerebral circulation in giant aneurysm surgery. AB - The surgical treatment of giant aneurysms usually requires temporary clipping of the aneurysmatic vessel. In planning the surgical approach and in applying temporary clips, the surgeon must consider collateral circulations. The functional integrity of the collateral vessels frequently decides the patient's outcome. In 8 patients with internal carotid artery giant aneurysm, measurements of blood flow velocities in the ipsilateral middle cerebral artery were performed preoperatively with transcranial Doppler ultrasound (TCD) during manual occlusion of the carotid artery at the neck. Three different perfusion patterns were established, and each collateral capacity was rated as insufficient, temporarily sufficient, or long-term unproblematic. Surgical strategies were conceived. In one patient with giant aneurysm of the middle cerebral artery the temporary occlusion test was not carried out preoperatively. Intraoperatively, collateral circulation was controlled using microvascular Doppler sonography (MVD). In 8 cases cortical blood flow (CoBF) was monitored by thermal diffusion flow probe and/or laser Doppler. In some cases, the complex pathological anatomy required a change in surgical strategy and a new MVD determination of collateral capacity. Despite these precautions 2 patients suffered ischemia of the basal ganglia and the white matter. PMID- 7478023 TI - The practitioner, the patient, the resident, and the pharmaceutical "gift". PMID- 7478022 TI - Treatment of severe intraventricular hemorrhage by intraventricular infusion of urokinase. AB - During the last three years, seven patients with severe intraventricular hemorrhage admitted to our clinic were treated with direct intraventricular infusion of urokinase. In each case, hemorrhage extended into the entire ventricular cavity and cast formation as well as an expansion of third and fourth ventricles were found. On the average, both the third and fourth ventricles became clear on the third day and the lateral ventricle on the ninth day after hemorrhage. Five of the seven patients showed good recovery or only moderate disability, and two died. Infection, convulsion, rebleeding, and peripheral or secondary hemorrhage due to the side effects of urokinase was not encountered during therapy. We conclude that this procedure can be applied effectively and safely in severe intraventricular hemorrhage. PMID- 7478024 TI - Clinical characteristics of delayed and immediate-onset combat-induced post traumatic stress disorder. AB - The aim of the current study is to examine the clinical characteristics of war related disturbances among veterans with delayed and immediate-onset post traumatic stress disorder (PTSD). One hundred twenty-five veterans who sought help for war-related disturbances, 8 years after the 1982 Lebanon War, filled out the PTSD Inventory, Impact of Event Scale, and SCL-90. Their scores were compared with those of 370 treated combat stress reaction casualties who filled out the questionnaires a year after the same war. Findings indicate that veterans from the delayed-helpseeking group suffer from a higher rate (92 vs. 59%) and a greater intensity of PTSD, more intrusive tendencies, and more severe general psychiatric symptomatology than those of the immediate-helpseeking group. These findings indicate that a fair number of combatants still seek help for war related disturbances almost a decade after the war. The complex relationship between delayed-helpseeking and delayed-onset PTSD is discussed. PMID- 7478025 TI - Barriers to screening: the theory of reasoned action applied to mammography use in a military beneficiary population. AB - BACKGROUND: Mammography practices of women eligible for military health care are not known. METHODS: A questionnaire was mailed to a stratified sample of 500 military beneficiary women. The hypothesis was that this group would not differ from the general population in regular mammography use (31%) and that the theory of reasoned action (TRA) is significantly associated with intent to obtain a mammogram. RESULTS: Only 12.8% of these women had regular mammography according to current recommendations. High participation is associated with breast cancer risk factors, income, education, perceived risk of breast cancer, and a doctor's recommendation. The expanded model of the TRA regression is significantly associated with intent. CONCLUSIONS: Regular use of mammography among military beneficiaries is lower than rates in the non-military population. Providers need to stress regular use and address scheduling problems and patient beliefs about mammography. The TRA is a useful model for screening behavior. PMID- 7478027 TI - Doxycycline for malaria prophylaxis in Australian soldiers deployed to United Nations missions in Somalia and Cambodia. AB - The operational effectiveness of daily doxycycline alone or combined with weekly chloroquine were assessed during deployments of Australian Defence Force personnel to malaria-endemic countries. Doxycycline was given as part of mandated disease prevention measures during United Nations missions to Somalia (900 men for 4 months) and Cambodia (600 men for 12 months over two annual rotations). In Somalia the soldiers were in an area of low endemicity and experienced only three malaria cases (one Plasmodium falciparum, two P. vivax), all occurring after returning to Australia. In Cambodia the level of malaria exposure varied greatly, resulting in eight malaria cases during the entire 2-year mission (two P. falciparum, six P. vivax). Doxycycline was generally well tolerated, with 1.7% (Somalia) and 0.6% (Cambodia) of the men requiring a change of medication to mefloquine due to adverse effects. Doxycycline is an effective chemoprophylactic agent during operational deployments when soldiers truly take it every day. PMID- 7478026 TI - Dietary fiber content of a six-day weighed military ration. AB - No studies have accurately reported on the total dietary fiber content of current military dining facility meals. The military nutrition data base is incomplete for total, insoluble, and soluble dietary fiber content. The Military Recommended Dietary Allowances has no recommendation for dietary fiber. The American Dietetic Association advises all Americans to consume 25 to 35 g/day. This study estimated total dietary fiber intake from a calculated analysis of a 6-day weighed military ration. The calculated fiber values for the sample military diet provided 6.8 g of total dietary fiber per 1,000 kcal. Substituting whole-grain cereal products for all the refined cereal products increased the fiber to 9.5 g/1,000 kcal. Calculated results were validated by chemical analysis. Total dietary fiber was 4.1 g/1,000 kcal for the sample military diet and 5.6 g/1,000 kcal for the whole grain test diet. Fiber content of the sample military diet was well below recommended levels but similar to that found in other studies on the typical American diet. PMID- 7478028 TI - Diarrhea morbidity among Dutch military service men in Goma, Zaire. AB - An outbreak of diarrheal disease occurring among service men of the Royal Netherlands Forces while serving in the refugee camps in Goma, Zaire, is presented. Post-travel questionnaires were given to 131 military service men returning from Goma. In addition, routine blood investigations were undertaken and stool examined for persons with abdominal complaints. A high prevalence (59%) of diarrheal disease associated with the eating of food served by a local hotel was noted. Fifty percent of persons who developed diarrhea in this period had symptoms persisting for up to 10 days. Pathogens were recovered from the stools of five persons, and four of these had Shigella organisms isolated. The outbreak of diarrhea among the service men was directly linked to the eating of food that was prepared by a local Goma hotel, the possible causative agent for this outbreak being Shigella organisms. Both military and civilian humanitarians working in areas such as refugee camps with a high prevalence of diarrheal diseases should carefully consider eating from local restaurants and hotels. PMID- 7478029 TI - The effect of simultaneous intravenous administration of nitroglycerin and heparin on partial thromboplastin time. AB - The purpose of this study was to determine whether intravenous nitroglycerin, when infused simultaneously through the same tubing with heparin, alters the anticoagulant effect of heparin as evidenced by partial thromboplastin time (PTT) values. A time series, cross-over design was used. The study included patients admitted to the Coronary Care Unit with a diagnosis of unstable angina. Patients were randomly assigned to one of two treatment groups: (1) heparin/nitroglycerin infused in the same site or (2) heparin/nitroglycerin infused in separate sites. After 18 hours, treatments were reversed. PTT results were measured seven times per subject. Ninety-eight subjects, 70 males and 28 females, participated. Mean age was 62.4 +/- 9 years. Using repeated measures analysis of variance, there was no change in the patients' PTTs (p = 0.47). Prior to this study, research was divided on whether nitroglycerin interferes with heparin's activity. This study indicates no significant difference in PTT in subjects receiving these infusions via separate or the same intravenous sites. PMID- 7478030 TI - Tick bites, clinical symptoms of Lyme borreliosis, and Borrelia antibody responses in Finnish army recruits training in an endemic region during summer. AB - Tick bites, associated clinical symptoms, and antibodies against Borrelia burgdorferi were investigated in 77 Finnish army recruits training during summer in an endemic region and 50 control recruits serving outside the habitat of ticks. During a follow-up of 6 months, 26.9% of the study recruits reported tick bites. None gave a history of erythema migrans. Five study recruits and none in the control group had a combination of two or three nonspecific symptoms compatible with Lyme borreliosis. Three of these five study recruits had been bitten by several ticks. Thirteen (16.9%) study recruits and two (4.0%) control recruits had positive antibody levels against B. burgdorferi in their first serum specimens. No significant change in immunoglobulin G antibody levels was seen between the first and second specimens. Immunoglobulin M antibody levels increased in the sera of 13 (11.9%) study recruits and 1 (3.0%) control recruit. We conclude that recruits training in tick habitats are at high risk of tick bites. All recruits starting their military training in endemic areas should be taught to recognize and remove ticks and advised to wear protective clothing. PMID- 7478031 TI - Soldier functioning under chronic stress: effects of family member illness. AB - The psychological functioning of soldiers with a handicapped child in the family were compared with that of soldiers without such a child through a survey of 443 soldiers. Self-report questionnaires were utilized to measure depressive symptoms, martial adjustment, social supports, stressful life events, military satisfaction, military performance, and coping. Differences between the 147 soldier-parents with a handicapped child and those without were examined using one-way analyses of variance. The results indicated that soldier-parents with a handicapped child showed significantly higher depressive symptoms, including lower scores on coping, less favorable perception of their military skills and abilities, and more pessimistic attitudes about their long-term military career options, than did the comparison group. Differences in marital satisfaction were not found. Also, perceived social supports played a significantly greater role in buffering the effects of stress on marital adjustment among families with a handicapped child than among those without. PMID- 7478032 TI - Laparoscopic lymph node dissection in the evaluation and management of patients with pelvic malignancy. AB - OBJECTIVE: This retrospective investigation describes our experience with laparoscopic pelvic and paraaortic lymph node dissection at a military medical center. METHODS: The charts of all patients undergoing laparoscopic pelvic and paraaortic lymph node dissection between July 1992 and May 1994 were reviewed for age, weight, cancer type, tumor stage, operative procedures, estimated blood loss, complications, number and pathologic results of lymph nodes removed, preoperative computed tomography findings, and postoperative therapy. RESULTS: The study group was composed of 20 patients, of which 15 had cervical cancer, 2 had ovarian cancer, and the rest had other pelvic malignancies. The mean age was 46 years old, with a mean weight of 155 pounds. The average number of lymph nodes removed was 26, with 9 of the 20 patients having positive nodes. Only one patient had preoperative radiographic evidence of lymphadenopathy. Mean estimated blood loss was 224 cc. Four patients required open procedures, two for bleeding during the laparoscopic node dissection and two who were unable to be completed through the laparoscope for technical reasons. Radical hysterectomy was avoided in two of the studied patients after finding positive lymph nodes. We conclude that in a military medical center, laparoscopic pelvic and paraaortic lymph node dissection in patients with certain pelvic malignancies is feasible and helps guide the management of these patients. PMID- 7478033 TI - Accessory bones of the foot: clinical significance. AB - In an effort to improve orthopedic clinic utilization, we reviewed the incidence of normal radiographic variants in the foot referred from the emergency room or outpatient clinic. The study was undertaken because normal radiographic variants are often confused with fracture. A retrospective study of all foot radiographs referred to the foot clinic at Madigan Army Medical Center was carried out. Distribution of normal radiographic variants and their confusion with fracture were evaluated. Thirty-six percent of all foot radiographs had identifiable accessory bones. The most common accessory bones in descending order were: os peroneum, accessory navicular, and os trigonum. All of the accessory bones identified in our study were asymptomatic. A thorough knowledge of location of normal accessory bones is mandatory and localization of foot symptoms by history and physical examination are necessary to facilitate correct diagnosis and treatment and to avoid unnecessary referral of these patients. PMID- 7478034 TI - Otologic features and teachings of a bombing attempt. AB - The authors present 13 casualties observed after the bombing attempt in Djibouti (March 1987). All of them suffered blast injury to the ear. The scene and the circumstances of the explosion are described. A correlation is sought between the location and position of each casualty and the lesions he presented, with reference to animal experiment data, namely the shock waves propagation conditions. The otologic lesions are commented on with reference to acute acoustic trauma, and the need for a special study of vestibular outcome of blast injuries is emphasized. PMID- 7478035 TI - A comparative study of digital images versus 35-millimeter images. AB - Digital imaging is an alternative to 35-mm Kodachromes and may be a useful tool to the physician. The purpose of this study was to compare the quality of digital images to 35-mm images and then compare the two methods based on the ability to make the correct diagnosis. Dermatology patients were photographed with both a digital camera and a 35-mm camera. Images from 22 patients were compared and graded on four criteria (sharpness of image, contrast, depth of field, and trueness of color). Diagnoses were then made from 30 pairs of images of different patients. In all graded areas the 35-mm images scored significantly higher. The correct diagnosis was made in 69.6% of the digital images compared to 77.3% of the 35-mm images, a difference that was not statistically significant. One may conclude that digital images hold promise for present and future use in dermatology and other areas of medicine. PMID- 7478036 TI - Three cases of adult attention-deficit hyperactivity disorder. AB - Three cases of adult patients with attention-deficit hyperactivity disorder who received their initial clinical evaluation and management in an Air Force Mental Health Clinic are presented. Diagnostic, therapeutic, and administrative issues pertinent to these patients are discussed. Clinicians should be alert to the diagnostic possibility of attention-deficit hyperactivity disorder in adult patients reporting the characteristic history and symptoms. PMID- 7478037 TI - Inflammatory pseudocyst associated with trisomy 21 and Hirschsprung's disease. AB - A 15-month-old black male with trisomy 21 and Hirschsprung's disease developed an inflammatory abdominal pseudocyst 1 month after Swenson pull-through repair. Although abdominal pseudocyst associated with ventriculoperitoneal shunts, peritoneal dialysis catheters, meconium peritonitis, and pancreatitis have been reported, this is the first report of a pseudocyst developing 1 month after a surgical repair of Hirschsprung's disease. PMID- 7478038 TI - Perforation in hypopharynx and deep cervical emphysema caused by blunt external trauma. AB - We present a case in which a blunt external trauma caused a perforating wound in the hypopharynx without any superficial damage on the skin of the neck. Deep cervical emphysema and general signs of systemic infection followed. The wound was sutured through an endoscope and the patient recovered uneventfully. The importance of the early diagnosis is emphasized. PMID- 7478039 TI - [Fibrinogen as a cardiovascular risk factor]. AB - Fibrinogen is a globulin of hepatic synthesis necessary for the coagulative process and is considered a risk factor of atherosclerosis disease. The increase in plasma fibrinogen seems to play an important role in the atherosclerosis disease and actually it represents a major, independent risk factor that should be considered for screening programs aimed at identifying individuals at high risk for cardiovascular disease. In the present review we'll propose to summarize current pharmacological knowledge, the relationship between fibrinogen and the best known cardiovascular factors like smoking and hypercholesterolemia. Furthermore we'll show the main mechanisms which make hyperfibrinogenaemia a pathogenetic factor of atherosclerosis. Lastly we analyse the possible fibrinogen lowering therapy. PMID- 7478040 TI - Myocardial protection by warm blood cardioplegia. A 31P nuclear magnetic resonance spectroscopy study of an isolated perfused rat heart. AB - OBJECTIVE: A study of the protective efficiency imparted by intermittent warm blood cardioplegia (WBCP) using perfused rat heart model. METHODS: Hemodynamic parameters were monitored simultaneously with metabolic changes in high-energy phosphates using 31P-NMR spectroscopy. Following 30 min perfusion with Krebs Henseleit (KH) buffer, all hearts were arrested for one hour (G1 and G2) and than reperfused with KH for 30 minutes. A warm oxygenated crystalloid cardioplegia (WCCP, modified St Thomas' hospital solution) was used for the control group (G1). The second group of hearts (G2) were arrested with oxygenated WBCP (K+ = 15 mM; Hct = 15-20%) and the third group (G3), was subjected to a protocol consisting of 4 periods (10 min each) of WBCP interspersed by 10 min of global ischaemia. RESULTS: The post-arrest percentage recoveries of LVDP, +dP/dt and HR were respectively: 88, 93 and 90 for G1 (n = 8); 97, 100 and 98 for G2 (n = 10); 76, 79 and 91 for G3 (n = 12). The corresponding metabolic recoveries of ATP and PCr were respectively, 85 and 90 for G1; 91 and 98 for G2 and 73 and 85 for G3. The PCr level declined during the arrest period in G1 contrasting with elevated PCr level (> 140%) during the WBCP arrest in G2. After an initial rise to approximately 140%, PCr level gradually decreased during the intermittent WBCP interval (G3). CONCLUSIONS: At normothermia, with equal CF rates, continuous WBCP provides better myocardial protection, through an effective oxygen supply, compared with WCCP. During the intermittent periods of ischaemia, certain metabolic and hemodynamic dysfunction occurs. PMID- 7478041 TI - Dipyridamole echocardiography and 99mTc-MIBI spect dipyridamole scintigraphy for cardiac evaluation prior to peripheral vascular surgery. AB - Coronary artery disease accounts for most of the early and late mortality and morbidity associated with vascular surgery. Cardiac pre-operative evaluation is mandatory for the assessment of cardiac risk. The aim of this study is to compare dipyridamole scintigraphy with 99mTc-MIBI (MIBI-dipy) and dipyridamole echocardiography (ECHO-dipy) and to evaluate their capability in identifying cardiac risk for strong events such as death, unstable angina or myocardial infarction. METHODS. Sixty consecutive patients (mean age 67 +/- 7) were enrolled. 52 performed ECHO-dipy, 51 MIBI-dipy. 40 patients went to aorto-femoral or aorto-iliac graft replacement and 15 to vascular angioplasty. Five patients did not undergo surgery. RESULTS. Eighteen patients (30%) had stress defects and 9 patients also rest defects with MIBI-dipy. Six patients new asinergic areas at ECHO-dipy. Three pts died in the first year follow-up for a cerebrovascular event, a myocardial infarction and a sudden death respectively. Sensitivity and specificity, positive and negative predictive value were 100%, 69%, 16%, 100% for MIBI-dipy and 66%, 94%, 40%, 98% for ECHO-dipy. CONCLUSIONS. As other authors reported, stress scintigraphy is a pre-operative test showing high sensitivity but with no satisfying specificity. Stress echocardiography, in our population, can produce a good negative predictive value. It is a less expensive and widespread clinical tool useful in the evaluation of preoperative patients. Its positive predictive power is not satisfying but it is shared with all non invasive pre-operative tests available now. PMID- 7478042 TI - [Echo-sclerotherapy in the management of varices of the lower extremities]. AB - Sclerotherapy has been used with satisfactory results, for several years in the treatment of varicose veins. Nevertheless sometimes sclerosis can be incomplete because of the morphology of lower limbs or because the varicose disease is not clinically evident. In addition, sclerotherapy can give rise to severe complications due to intrarterial or extraluminal injections. In order to exceed this limits, some authors suggested to use a new technique, the echosclerotherapy, which was presented for the first time in Strasburg 1989 by Knight and Vin. Echosclerotherapy is a good help for traditional sclerotherapy, especially when it is applied in the sclerosis of the short saphenous veins, of perforating veins or in unfavourable anatomical situations. From May to November 1993 at the Second Surgical Department of Padua University, 31 patients, 29 women and 3 men, have been treated by echosclerotherapy. 25 patients had great saphenous varicose veins; 3 patients had varices due to perforating veins of the popliteal fossa and 3 patients varices due to Hunter perforating veins. In 48.4% of cases we obtained a complete sclerosis of the vessel; in 38.7% a stump remained near the sapheno-femoral junction of about two centimeters; in one case the treatment was not completed and in one case remained a stump of ten centimeters. Only in two cases Echosclerotherapy was not able to obtain sclerosis. None of the patients had major complications and nobody had deep vein thrombosis. If we consider our results altogether we can say that in 87% of cases we had good results.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7478043 TI - [Ambulatory angiologic diagnosis using the Doppler c.w. (continuous wave) method]. AB - A one year clinical activity of instrumental angiological diagnostics (doppler c.w. examinations) was evaluated and analysed. 970 outpatient continuous wave doppler have been performed: 37% examinations of epiaortic vessels (TSA), 31% examinations of lower limb veins, 28% examinations of lower limbs arteries. The question has been inappropriate (normal exam) in 81% of TSA, in 67% of AIA and in 52% of AIV. The authors think that the reasons for these results are a large misinformation from the physicians postulant and a consequent abuse of the instrumental method. It's therefore necessary to put a clinical filter to regulate the access to the instrumental (high technological) examinations, too often unnecessary. PMID- 7478044 TI - [Prevention of vascular intimal hyperplasia in small caliber prostheses. Preliminary results]. AB - Intimal hyperplasia is one of the main risk factors for the patency of small diameter bypass grafts. The standard unfractioned heparin (UH) is able to control this phenomenon but the clinical use is not fit for long term treatment; on the other hand the antiplatelet drugs have an anti-thrombotic effect but they seem to be unable to control intimal hyperplasia. Low molecular weight heparins (LMWH) have an anti-thrombotic effect superimposable to that of UH with minimal side effects and might inhibit intimal hyperplasia too. Based on these criteria, we carried out an experimental study on sheep with the aim of evaluating the efficacy of postoperative treatment with LMWH versus an anti-platelet drug in controlling intimal hyperplasia and growth of true endothelial cells in small prosthetic ePTFE grafts (4 mm) interposed in the carotid artery. At the operation, 30 sheep were randomly located in 3 groups: A = control group, no treatment; B = Ticlopidine hydrochloride 250 mg/bid by mouth for 4 weeks; C = LMWH 3.075 IU AXa (0.3 ml) sc preoperatively and then once a day for the same period. Complete thrombosis of the graft occurred in 7 sheep of group A, 5 in B and 2 partial in group C. The intimal hyperplasia was moderate-severe in group A, mild-moderate in group B and no-mild in group C. True endothelial cells were found mainly in the LMWH group; in the other groups and in other portions of the grafts the cellular coverage was accomplished almost completely by fibroblasts. The study is still in progress with 6 further sheep treated with LMWH. PMID- 7478045 TI - [Lymphedema. New non-invasive methods for diagnosis and follow up]. AB - Lymphedema is still a difficult clinical problem, poorly investigated and new methods of evaluation are needed to improve the understanding of its pathophysiology. Lymphoscintigraphy is diagnostic but cannot be repeated frequently in the follow-up. In this study we have evaluated four new methods of evaluation of lymphedema which may be used to quantify the problem and to follow up patients. These methods are: A. the evaluation of the ratio between the concentration of lymphatic fluid proteins and plasma proteins concentration (CL/CP); B. the test of the spontaneous clearance of a haematoma; C. high resolution ultrasound imaging of low density spaces (SBD) in the subcutaneous tissue, possibly corresponding to dilated lymphatic spaces; D. the combination of imaging and CL/CP ratio. Comparable groups of normal subjects, patients with primary lymphedema diagnosed with lymphoscintigraphy and patients with chronic venous insufficiency have been evaluated. The four methods appeared useful to differentiate normal subjects from those with lymphedema. However the separation between lymphedema and chronic venous insufficiency was less evident. Edema due to systemic causes (cardiac failure, nephrotic syndrome or chronic venous insufficiency) is also differentiated from lymphedema. In conclusion these tests may be useful to evaluate lymphedema (particularly in the early phases, when the clinical presentation is unclear), to follow-up its evolution and possibly to evaluate the effects of treatments. PMID- 7478046 TI - [Myointimal hyperplasia in peripheral artery reconstruction]. PMID- 7478047 TI - [Arrhythmogenic right ventricular dysplasia. Report of a familial case]. AB - Arrhythmogenic right ventricular dysplasia is a pathologic condition of unknown origin that primarily affects the right ventricle free wall characterized by progressive atrophy with fibrous-fatty substitution of the myocardium. The patient affected may present in childhood or as an adult with cardiomegaly or/and ventricular arrhythmias with a left bundle branch block configuration. The reports of familial occurrence suggest the etiologic role of a genetic defect with autosomal dominance and variable expression and penetrance, an alternative, myocarditis (toxic or infectious) is a possible cause. The aim of the present study is an non-invasive instrumental evaluation of the family of a young patient affected by arrhythmogenic right ventricle dysplasia. The authors evaluated 10 patients, 7 women and 3 men, all asymptomatic, with electrocardiography, exercise stress test on a bicycle ergometer, 24-hour Holter monitoring and two-dimensional echocardiography. A patient had certain ARVD diagnosis, while in the other 9 patients all the instrumental investigations were normal. This observation seem to confirm that ARVD may be familial and justify a systematic in-depth familial study in all demonstrated cases of ARVD and the follow-up of all patients identified who had arrhythmias or isolated right ventricular morphologic anomalies. PMID- 7478048 TI - Multivariate prediction of in-hospital mortality associated with surgical procedures. AB - BACKGROUND: The aims of this prospective multicenter study were to identify variables associated with in-hospital mortality among patients undergoing surgical procedures, to develop a prediction rule, and to statistically validate its reliability. METHODS: Data from 24,654 consecutive informed patients over 15 years of age were collected from 22 surgical centers between January 1989 and December 1990. Using logistic regression analysis separate models were fit for seven surgical disciplines to predict the risk of 30-day in hospital mortality. Variables used to construct the regression models included age, sex, systolic blood pressure, renal dysfunction, hepatic dysfunction, concomitant diseases, severity of surgery, priority of surgery and duration of anesthesia. The performance of the prediction rule was evaluated by computing sensitivity, specificity and predictive values, analyzing the ROC curve and comparing observed with expected deaths. RESULTS: The significance of the independent variables varied within each model. All models significantly predicted the occurrence of in hospital mortality. At a 0.5 cuptoint of predicted risk sensitivity of prediction rule was 99.89%, positive predictive value 98.51%, and overall predictive value 98.41%, whereas specificity was 7.92% and negative value slightly higher than 50%. The area under the ROC curve was 0.80 (perfect, 1.0). The correlation between observed and expected deaths was 0.99. CONCLUSION: This prediction rule, developed using multicenter data, is characterized by the following advantages: includes only nine variables; can be utilized by seven different surgical disciplines; is highly accurate, and is easily available to clinicals with access to a microcomputer or programmable calculator. This validated multivariate prediction rule would be useful both to calculate the risk of mortality for an individual surgical patient and to contrast observed and expected mortality rates for an institution or a particular clinician. PMID- 7478049 TI - [Cardiocirculatory stability and early extubation in patients undergoing thoracic surgery with propofol as the single anesthetic agent]. AB - OBJECTIVE: To determine the efficiency of total intravenous anesthesia technique (TIVA) using propofol on maintaining cardio-circulatory stability and rapid recovery time in patients undergoing thoracotomy. DESIGN: Prospective study. SETTING: University General Hospital in Spain. PATIENTS: Twelve patients, undergoing elective thoracic surgery, including lung resection with one-lung ventilation with more than 120 minutes of anesthetic time. METHODS: ECG, blood pressure (BP), heart rate (HR), pulse oximetry (SpO2), end tidal CO2 (ETCO2) and muscular relaxation with Relaxograph were monitored. Anesthesia was performed with propofol 1.5-2.5 mg/kg i.v. and maintained with continuous propofol infusion 4 +/- 1.4 mg/kg/h. Fentanyl, 10 mg i.v., was administered in bolus. After succinylcholine 1 mg/kg i.v. or atracurium 0.5 mg/kg i.v., trachea was intubated with a double-lumen tube or a standard cuffed endotracheal tube. The patients were ventilated with air-oxygen mixture. Maintenance of muscular relaxation was obtained with atracurium 0.2 mg/kg i.v. RESULTS: No significative differences were registered in the values of heart rate, SpO2, ETCO2 and between intraoperative blood pressure and the postextubation blood pressure values. Recovery time, from discontinuation of the propofol until postoperative extubation, was 8.25 +/- 4.8 minutes. CONCLUSIONS: Performing a TIVA with propofol good cardiocirculatory stability was obtained. Continuous monitoring of relaxation, graphically recorded, knowing the bolus rate of muscle relaxant, allows us to antagonize it totally, facilitating patient recovery and its early postoperative extubation. PMID- 7478050 TI - [Comparison of 2 modified Fick methods and thermodilution for determining the cardiac output in patients with mechanical ventilation]. AB - OBJECTIVE: To evaluate the effectiveness of two cardiac output measurement methods based on a modified Fick equation, that calculate the O2 consumption (VO2) dividing the CO2 production (VCO2) by a fixed respiratory quotient (RQ). DESIGN: Comparative study. SETTING: One 5 beds general intensive care unit in a 500 beds general hospital. PATIENTS: Ten mechanically ventilated critically ill patients requiring the right heart catheterization. The mean age was 65.5 years and the mean APACHE II score at admission was 24. MEASUREMENTS: The cardiac output was measured using two modified Fick methods. The VO2 was calculated dividing VCO2 by 0.9, while the arteriovenous O2 content difference was calculated using the mixed venous O2 content in the first method (COF), and the central venous O2 content in the second one (COFra). Moreover the cardiac output was measured with the thermodilution technique (COTD) as gold standard. RESULTS: The mean difference between the COTD and COF determinations was 0.571 L +/- 1.81 L, with limits of agreement ranging from -3.057 to +4.200 L. The mean difference between the COTD and COFra determinations was -0.322 L +/- 2.05 L, with limits of agreement ranging from -4.430 to +3.785 L. Both differences were nonsignificant. The correlation coefficients with COTD were: COF determinations 0.72, COFra determinations 0.70. In the group of COFra determinations less than 7 L the mean difference between COTD and COFra was 0.495 L with limits of agreement ranging from +2.208 L to -1.218 L. CONCLUSIONS: The correlation coefficients of the two modified Fick methods with COTD are good, and the mean differences between their results and the gold standard are small, but the low precision of both tested methods demonstrated by the very large limits of agreement, severely reduce the clinical reliability of the measurements. Only for the less than 7 L cardiac outputs the COFra limits of agreement with COTD are narrow enough, and in this range the technique can be useful e.g. revealing a low cardiac output. PMID- 7478051 TI - [Ultrasonography-guided identification of the lumbar epidural space]. AB - The study was designed to assess the reliability of sonographic evaluation in the prediction of the depth of the lumbar epidural space. Forty males, scheduled for epidural anesthesia for surgical repair of inguinal hernia, were prospectively studied. Patients were placed in a sitting position and sagittal scanning of the lumbar spine was performed with a 5-MHz transducer over the fourth or fifth interspace in order to identify the deeper hyperechogen interface, which represents the landmark between the ligamentum flavum and the epidural space. Ultrasound depth was measured and transducer removed. A Tuohy needle 18 G was then introduced percutaneously according to the standard technique and a rubber slide placed over it, so that it depth of insertion could be accurately measured. The potential for using ultrasounds for prediction of the distance from skin to epidural space was analyzed using a simple linear regression analysis; p values < 0.05 were considered significant. Mean values of ultrasound depth and needle depth were respectively 51 mm (SD 6.3) and 50.9 mm (SD 6.2); the correlation coefficient was 0.99. Ultrasound scanning of the lumbar spine provides an accurate measurement of the depth of the epidural space, which can facilitate the performance of the epidural anaesthesia and may decrease the complication rate, particularly in those patients in which anatomic landmarks are obscured. PMID- 7478052 TI - [Anesthesia in experimental single-lung transplantation: hemodynamic and respiratory changes during prostaglandin E1 and nitroglycerin infusion]. AB - INTRODUCTION: The haemodynamic and oxyforetic modifications induced by continuous infusion (c.i.) of Prostaglandin E1 (PGE1) and Nitroglycerin (NG) during experimental lung transplantation have been studied. METHODS: Twenty lung transplants have been performed on pigs (Landrace). Haemodynamic monitoring consisted of ECG, femoral cannulation for arterial pressure and Swan Ganz catheter. Continuous infusion of drugs started during pneumonectomy. Data have been collected in phase 1: after the induction of anaesthesia; in phase 2: 30 min after the start of pulmonary vasodilation in one lung ventilation with left pulmonary artery clamped; in phase 3: 30 min after lung revascularization; in phase 4: at the end of the surgery. The animals have been divided in two groups. Group A consisted of 10 cases treated with 50 ng/kg/min in phase 2 and 25 ng/kg/min in phase 3 and 4 of PGE1; group B consisted of 10 cases treated with 1 ncg/kg/min in phase 2 and 0.5 ncg/kg/min in phase 3 and 4 of NG. RESULTS: In phase 2 and 4 PA, RVSWI, PRVI increased in group B more than in group A (p < 0.05); in phase 3 PA, PAWP increased in group B more than in group A (p < 0.05). DISCUSSION: In our experience PGE1 has shown to be a stronger pulmonary vasodilator than NG. PGE1 can be considered the drug of choice for pulmonary vasodilation in experimental single lung transplantation, even if more investigations are needed for reaching the real goal of this procedure. PMID- 7478053 TI - [Rectal administration of anesthetic agents]. AB - OBJECTIVES: To collect data in the current literature dealing with the diffusion, the reliability and the effectiveness of the rectal administration of anaesthetic drugs. To evaluate differences with parenteral administration. DATA SOURCES: Pharmacokinetics and clinical studies published in recent years in indexed journals. STUDY SELECTION: Based on the study methodology, drugs employed and pharmacokinetic parameters evaluated. DATA EXTRACTION: Factors involved in absorption of drugs from the rectal mucosa, clinical effect and pharmacokinetic data of the following drugs: diazepam, flunitrazepam, midazolam, ketamin and methohexital, then a brief evaluation of other drugs: thiopental, etomidate, morphine and chloral hydrate. DATA SYNTHESIS: The most widely used drugs are benzodiazepines: they are safe, easy to manage and highly effective; among them midazolam has the best kinetic and dynamic pattern. Ketamin is useful during painful diagnostic procedures; with the use of barbiturates there is a greater risk of respiratory depression and more caution must be employed. CONCLUSIONS: Wide intervariability of rate of absorption, achievement of plasma levels and clinical effect is a relevant drawback of this technique, such to make it not preferable to the parenteral route, when both are feasible. It deserves, anyway, more consideration, and maintains its validity for the preparation of the paediatric patient to general anaesthesia. PMID- 7478054 TI - [Impact of DRG on the American hospital system. Development of day hospital]. PMID- 7478055 TI - [Role of totally implantable systems for long-term vascular access in the treatment of the neoplastic patient]. AB - OBJECTIVE: To assess retrospectively the effectiveness, the safety and the impact on the quality of life of the Subcutaneous Infusion Ports (SIPs) for prolonged venous access in the treatment of neoplastic patients. DESIGN: Retrospective analysis of a case series collected-during 30 months (April 1991-September 1993). SETTING: General Surgery Division and Radiochemotherapy Service. PARTICIPANTS: 35 patients (23 male, 12 female), aged between 27-80 years, received 37 SIPs. 2 patients received 2 SIPs. 34 patients were affected by neoplasm; 1 patient had short gut syndrome secondary to massive small bowel resection. INTERVENTIONS: The SIP was implanted in all patients in the operating room with the Seldinger technique ("catheter over wire"). The suclavian vein, through an infraclavicular approach, was the only site of venous access. One SIP was implanted in the hepatic artery. All neoplastic patients received chemotherapy. The patient with short gut syndrome had parenteral nutrition. An external infusion device (CADD-1, CADD-plus, Pharmacia) was used in 13 patients for the continuous infusion of either 5-Fluoro-Uracil (1000 mg/m2/die for 5 days, every 28 days) or 5-Fluoro Deoxy-Uridine (0.15-0.30 mg/kg/die for 14 days, every 28 days). SIP management required the port "flush" with normal saline solution with heparin (100 U/ml) at least every 40 days, using Huber needle. All patients were treated as outpatients. MAIN OUTCOME MEASURES: All intraoperative and long-term complications, recorded in the charts of the patients, were reviewed. The rate of intraoperative complications was assessed over 37 procedures. Long-term complications were assessed over 28 patients (7 patients were excluded for lack of follow up data). Incidence of complications was analyzed over patient days, considering the cumulative permanence time of the SIP in a single patient. RESULTS: We recorded 4 intraoperative complications (10.8%). None of them required to stop the procedure and to delay the implant of the SIP. During the follow-up period (range 1-18 months) we had 1 major complication (pleural effusion secondary to TPN extravasation), that is 3.5% of the patients (1/8255 patient days), and 3 minor complications (10.7% of the patients; 1/2751.6 patient days). Patient tolerance was good in all cases. CONCLUSIONS: SIP is a safe and effective device for prolonged venous access. We stress the need of a wider use of this type of device for the management of neoplastic patients. PMID- 7478056 TI - [Surgical treatment of vascular injuries of the legs]. AB - This retrospective study was carried out on 24 patients admitted to the Surgical Thoracic Department of the United Hospitals in Foggia between 1984 and 1992. In most cases are patients with multiple traumas, victims of road accidents. Different types of surgical treatment have been used, from the ligation or suture of the perforating artery to the resection with anastomosis T-T or interposition of graft. When there were bony injuries too, the vascular repairing venous arterial, was carried out after the orthopedic one. The nerves have been repaired after the vessels. The nervous injuries are the chief cause of permanent functional damage, because today the success of the surgical operation used in the vascular injuries of the limbs is increased to values about 90%. For the vascular sutures we have used material that is not absorbed, the monofilament. The present average rate of amputation of 10% can be more lowered if we put attention to complementary expedients and if there is a more active and quicker collaboration among the different specialized departments. PMID- 7478058 TI - [Assessment of sapheno-femoral junction continence in 42 patients with primary varicocele]. AB - Varicose veins present a major health problem, causing chronic leg symptoms and disability to a large segment of population. The prevalence rate in adults shows great geographical variations and increases with age. Several authors reported that most cases of primary varicose veins are due to valvular incompetence of saphenofemoral junctions. On the base of close physiopathologic and hemodynamic relationships between primary varicose veins and primary varicocele we decided to evaluate the continence of saphenofemoral junctions in a selected group of young men affected by primary varicocele. MATERIALS AND METHODS. 42 patients were included in the study. The age ranged from 13 to 55 years; the mean age was 28 years. In all cases a clinical and instrumental (ultranosography. Doppler flowmetry) diagnosis of primary varicocele was performed. Bidirectional Doppler flowmetry was used to evaluate the continence of 84 saphenofemoral junctions. RESULTS. Among the group of studied 36 (85.7%) showed a monolateral or bilateral valvular incompetence of saphenofemoral junctions; in the remaining 6 patients (14.3%) there was no evidence of valvular insufficiency. In 84 saphenofemoral junctions studied 52 (61.9%) were found incompetent, 32 to the left side and 20 to the right side. Moreover 2 patients had clinically detectable varicose veins of the lower limbs and symptoms related to chronic venous insufficiency. DISCUSSION. Varicose veins are one of the most prevalent chronic conditions in industrialized countries and places a considerable demand on the health services. Since this pathology is potentially preventable it seems very important to select within populations asymptomatic groups in which there is an increasing risk of chronic venous insufficiency and to perform a clinical and instrumental assessment, based on non invasive techniques. In the past varicose veins were related to other diseases and among them some authors suggested primary varicocele, that is due to venous spermatic reflux and/or valvular incompetence; furthermore varicocele and varicose veins are often familiar. In our 42 patients affected by varicocele only 6 (14.3%) had no Doppler flowmetry evidence of incompetence of saphenofemoral junctions and more than half (61.9%) of the junctions were found incompetent. CONCLUSIONS. In our experience we found a considerable relationship between primary varicocele and venous incompetence of saphenofemoral junctions. Perhaps these patients must be considered a group with an increasing risk to develop primary varicose veins. So we suggest that clinical examination and instrumental assessment should be performed in young men affected by varicocele, in order to obtain a prevention of chronic venous insufficiency of the lower limbs. Nevertheless this retrospective study has to be further confirmed by longterm prospective studies. PMID- 7478057 TI - [Clinical and hemodynamic effects of external valvuloplasty of the sapheno femoral junction]. AB - Selected patients with primary varicose vein disease and sapheno-femoral reflux as the only point of regurgitation in the affected lower limb and duplex evidence of mobile valve leaflets underwent external valvuloplasty of the sapheno-femoral junction. Operations were performed under local anesthesia in one day-surgery. One PTFE sleeve 0.4 mm thick, in average 1.2 cm long and able to uncircle a circumference in average of 1.7 cm was put around the terminal Long Saphenous vein valve, located in the last centimeter of this vein. If also the subterminal Long Saphenous Vein valve site, located 5 cm below is dilated a second valvuloplasty was performed. In 15 case we used intra-operatory video-angioscopy guide. This examination allowed us to exclude two patients with valve damages not duplex demonstrable. Furthermore angioscopy permitted immediate demonstration of restored valvular function. Follow-up lasted on average 48 months: two early postoperative long saphenous vein thrombosis, and two late sapheno-femoral reflux recurrences were the main complications. We never observed graft infection. AVP was reduced and RT, measured by the means of LRR, had a prolongation after surgery. Both measurements had an highly significant difference from a statistical point of view, comparing pre and postoperative values. 95% of the operated patients have stable varices reduction. Long Saphenous Vein patency was recorded in 37 cases (92.5%). On the contrary, after high ligation, the fate of the saphenous vein in 21% of case is the occlusion. Symptoms of venous insufficiency disappeared completely in 80% of cases and improved in 95% of the operated patients.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7478059 TI - [Color Doppler ultrasonography in the diagnosis of solid breast lesions]. AB - The color-Doppler signals in 50 patients with breast lumps were assessed subjectively. Number and location of vessels within the masses were recorded and compared with histologic diagnosis and lesion size. All the malignancies were vascularized but only 16/37 of the benign ones showed vessels. All malignancies but one displayed > or = 2 vessels, which on the other hand occurred in only 4/16 vascularized benignancies. There was a relation between size of the lesion and number of the observed vessels, and color-Doppler proved more reliable in the differential diagnosis of breast masses when lesions less than 2 cm were considered. PMID- 7478060 TI - [Psychological assessment of patients who have ++undergone breast reconstruction using 2 different technics: autologous tissue versus prosthesis]. AB - Breast reconstruction has become an available option for most patients undergoing mastectomy: in fact many authors agree that breast reconstruction does not interfere with possible therapies and improves the quality of life of women. The aim of the study was to evaluate the psychological adjustment of patients who had immediate or delayed reconstruction using 2 different methods: implants and autologous tissues. The study population (115 patients) was derived from patients who underwent breast reconstruction in the period January 1988-December 1991, in follow-up at the Department of Plastic and Reconstructive Surgery; no patient was undergoing psychological therapy. 58 patients underwent breast reconstruction using implants and 57 using Transverse Rectus Abdominis Myocutaneous Flap (TRAMF). Informations were gathered, including the patient's age, the number of offspring, the marital status, the scholastic education, the job and the relapse between mastectomy and reconstruction. The psychological instruments consisted in three standardized self-administered questionnaires: Psychological Distress Inventory (PDI), State Trait Anxiety Inventory form Y (STAI), Eysenk Personality Inventory (EPQ-R). These tests were chosen to gauge the psychological distress, such anxiety, anger, depression and psychosocial maladjustment. To better perform the changes of body image after breast reconstruction, women were requested to answer three more specific questions about the sexual desire, physical image and social relationships. The 102 patients assessed in this study indicate low incidence of psychological distress and adaptive coping strategies. Impairment was reported, regarding body image, by patients undergoing delayed reconstruction; in these patients higher scores in distress tests were observed. PMID- 7478061 TI - [Predictive value of ultrasonography in the study of aortic abdominal aneurysms]. AB - Several authors have collected a great deal of data in the study of abdominal aorta aneurysm through the swift diffusion of ultrasonography in the abdominal diagnostics. The authors illustrate their experience confirming the excellent reliability of ultrasonography in the diagnosis of abdominal aorta aneurysms with respect to other methods of imaging (Angiography, CAT, etc) not with a view to replacing them but as a useful means of screening. PMID- 7478062 TI - [Effects of gastric resection by the Billroth II technic on reflux esophagitis associated with duodenal or pyloric ulcer]. AB - Although duodenal ulcer and reflux esophagitis may frequently co-exist, it is not clear whether surgery for duodenal ulcer associated with reflux esophagitis should not be associated with anti-reflux plastic surgery. The authors make a retrospective evaluation of the prevalence of reflux esophagitis (endoscopic diagnosis) in a consecutive series of 633 patients undergoing elective gastric resection according to Billroth II for duodenal or pyloric ulcer during the period 1974-1992 and assess the effects of surgery on co-existent esophagitis. The prevalence of reflux esophagitis associated with duodenal or pyloric ulcer was 12.3%. In patients in whom stenosis represented the indication for ulcer surgery, the prevalence of esophagitis was 20%, whereas it was 5% in those operated because of failure to respond to medical therapy of hemorrhage (p = 0.000001). In 95% of patients, controlled endoscopically 6 months after surgery, Billroth II gastric resection led to the resolution or improvement of associated esophagitis. The authors conclude that by eliminating the main pathological factors of reflux esophagitis associated with duodenal ulcer (hypersecretion of gastric acid, impeded gastric emptying) gastric resection is sufficient to achieve the resolution of esophagitis. PMID- 7478063 TI - [Jejunal diverticulosis: report of a case and review of the literature]. AB - The jejunal diverticula are a rare pathology, they are verifiable with an incidence from 0.06 to 1.3% autoptical reports. Furthermore, only 10-40% of them are symptomatic. Nevertheless, in this case the beginning is often acute because of a perforation or a hemorrhage. In both cases the medical therapy is associated with a mortality rate which is too high to be acceptable, so therefore, emergency surgery is the only possible choice. The authors present a clinical case resulting from their observation with an acute beginning, this case is used as a starting point for an accurate review of the international literature on the subject. The purpose of this study is to contribute to a correct understanding and approach to the problem, above all through an exact preoperative diagnosis so that unnecessary delays in surgery may be avoided when a patient's life is at risk. PMID- 7478064 TI - [Carcinoid of Meckel's diverticulum]. AB - The carcinoid of Meckel's diverticulum is a very rare lesion, and about 100 cases in the literature have been described. Most of these tumors are casually find out at autopsy or laparoscopy for other pathology. Seldom was described a sintomatology correlated to carcinoid of Meckel's diverticulum presence, except the rare case where was formed a carcinoid syndrome. Authors present a clinical case where was casually found by a carcinoid of Meckel's diverticulum during an appendicectomy. The intention is to underline the importance of research the Meckel's diverticulum every time, and in particular in case of young subjects, remove it and perform an histological valuation, to exclude the presence of a carcinoid that could give metastases to local lymph-nodes or liver. PMID- 7478065 TI - [Invaginating colonic lipoma as a cause of intestinal occlusion]. AB - The authors describe three cases of single invaginating colon lipoma, causing ileus. They review the literature and make some remarks. PMID- 7478066 TI - [Unusual case of reappearing biliary ileus in a cholecystectomized patient with late stenosis of ileo-ileal anastomosis]. AB - An unusual case of alternating jejunal obstruction, caused by a former latero lateral anastomosis stenosis, complicated by two stones into the afferent jejunal tract bottom, is described. The anamnestic data of previous cholecystectomy and operative finding of no one biliary fistula clue, make the authors doubtful about a hypothetic calculi migration: it is likely calculi are formed into the bowel. PMID- 7478067 TI - [Dieulafoy ulcer associated with acute drug-induced gastritis: a rare case]. AB - The lesion in Dieulafoy's disease consists of an unusually large and tortuous artery in the base of an ulcer. The disease is underdiagnosed rather than truly rare and often presents with a massive and potentially fatal, usually repeated, haematemesis. The authors describe a case of Dieulafoy's disease associated with acute drug-induced gastritis whose gravity required, in the case reported, a very serious emergency treatment. PMID- 7478068 TI - [Nephro-jejunal fistula associated with nephro-cutaneous fistula]. AB - The development of a renal fistula is a rare event which results from an acute or chronic inflammatory process in the kidney. The authors report a very rare case of nephrojejunal and nephro-cutaneous fistulas due to acute and chronic pyelonephritis in a patient with staghorn stones of the left kidney. PMID- 7478069 TI - [Free tension in inguinal hernioplasty]. AB - The authors report their experience of a new surgical technique for inguinal hernia repair. This operation first introduced by Lichtenstein is accurately described by the authors. This kind of hernia repair was introduced in our operative unit because we think that the first cause of recurrence is the approximation of normally unopposed tissue under tension. This concept permits hernia repair without any suture line tension. For this principle the authors have been employing for one year the tension-free herniolplasty technique that was described by Lichtenstein. The authors consider a preliminary report of 144 cases in view of the long term follow-up; however, the results to date have been extremely satisfying: no infection has happened, no painful drugs was used, local anesthesia was usually employed, thus allowing intraoperative testing; early recovery. Finally, the technique is simple, rapid, relatively less painful, safe and effective. PMID- 7478070 TI - [Lateral calcaneal flap in the reconstruction of the calcaneal and malleolar region]. AB - The authors discuss the efficacy of the lateral calcaneal artery flap in repairing the calcaneal and malleolar regions. The lateral calcaneal artery flap is a safe flap, with constant vascularization represented by the lateral calcaneal artery and the external saphenous vein; the flap has a sensory nerve (the sural nerve) and is a sensory flap. Its use in the calcaneal and malleolar region is very safe, because the vascular pedicle is constant and surgical transfer easy. PMID- 7478071 TI - [Is the ligation of the hepatic artery in the emergency treatment of liver injuries justified?]. PMID- 7478072 TI - [Amyloidosis: state of the art]. AB - Amyloidosis is characterized by a heterogeneous group of diseases due to deposition of a fibrillar, proteinaceous material, in different tissues. In this review pathogenetic mechanisms, clinical and therapeutic findings of the most important aspects of the disease, are described. PMID- 7478073 TI - [Effects of simvastatin on blood levels of lipoprotein (a)]. AB - The efficacy of Simvastatin to reduce plasma cholesterol is well documented. Other molecule within the lipo-lipoprotein family, such as, particularly, lipoprotein (a) -Lp(a)-, have been recently found to have a predictive and/or causative role in atherosclerosis. Based on the above consideration, we studied 20 patients (7 females and 13 males), mean age 52.4 +/- 14.2 years, affected by primary hypercholesterolemia to evaluate the effect of simvastatin on Lp(a), in addition to the classic lipidic parameters. Five weeks after suspension of lipid lowering drugs and on a normal caloric-fat diet, were given 20 mg simvastatin/day for 12 months. Clinical and laboratory parameters, cholesterol (CH), triglycerides (TG), high density and low density lipoprotein cholesterol (HDL-CH and LDH-CH) measured enzymatically, apoproteins A1, B measured radial immunodiffusion technique and Lp(a) measured as apoprotein(a) with immunoradiometric assay and were evaluated before therapy and after 12 months of therapy. Simvastatin determined a significant reduction in total cholesterol and cholesterol-LDL (CH 327.7 +/- 44.4 vs 255.5 +/- 37.3, p < 0.0001; LDL-CH 257.1 +/ 60.9 vs 183.8 +/- 46.9, p < 0.0001) and a significant increase in HDL-CH (36.7 +/- 5.9 vs 40.2 +/- 5.7, p < 0.005); no variation was observed in triglycerides (TG) levels. Simvastatin therapy further determined a significant increase in Lp(a) plasma levels (43.8 +/- 25.6 vs 50.5 +/- 28.0, p < 0.02). The our data, in agreement with those documenting the beneficial effect of Simvastatin in greatly decreasing CH and LDL-CH, but point out the need for further studies concerning the long-ter effect of simvastatin on Lp(a), in order to fully establish its role in the secondary prevention of atherosclerosis. PMID- 7478074 TI - [Extrahepatic manifestations of HCV infection]. AB - It is well known that hepatitis C virus (HCV) can replicate outside the liver, particularly in the blood mononuclear cells. This fact may account for the systemic involvement, primarily due to immune-mediated mechanisms, which is commonly observed in patients affected with chronic hepatitis C. In this review the main extra-hepatic manifestations of the HCV infection are described and classified, according to the clinical involvement, namely dermatological, haematological, rheumatological, endocrinological, ophthalmological ad respiratory syndromes. In particular, the well-established causative role of HCV in determining certain clinical disorders, i.e., mixed cryoglobulinaemia and porphyria cutanea tarda, where the anti-HCV seroprevalence is higher than 70%, is emphasized. Moreover, some problems relevant to the diagnosis of the HCV infection in a proportion of autoimmune diseases (for example, primary Sjogren's syndrome), are discussed. Indeed, in these clinical disorders, hypergammaglobulinaemia, which represents an interfering factor with the results obtained by the screening anti-HCV tests, is a typical serologic feature. PMID- 7478075 TI - Zinc and immune function. AB - Zinc and immune function relationship has been extensively studied. Both in experimentally induced mineral deficit and in genetically determined deficit observable in acrodermatitis enteropathica and in enteropathy of Danish A-46 cattle, a B and T dependent antibody response decrease, a T dependent cytolytic response decrease and a natural killer cytotoxic activity decrease are present noteviously. Serious reduction of the immune function is present, in proportion to the value of low zinc plasmatic level, in elderly patients, in malnourished and seriously zinc deficient children, in patients subjected to total parenteral supply, in HIV infections and especially in evident AIDS: in this condition the plasmatic zinc level can be considered, together with the CD4+ lymphocytes amount and the B2-microglobulin value, a disease progression marker. Zinc immunostimulating action mechanisms are complex, although thymic hormone (of which zinc is essential cofactor) stimulation seems to be most important. Zinc supplementation, also parenterally, can be useful in immunodeficiency (in the elderly, in the post-surgical patients, in genetically determined or alimentary induced deficit, in AIDS. PMID- 7478076 TI - [Meningeal carcinosis: early clinical manifestations of inflammatory cancer of the breast]. AB - Inflammatory carcinoma of the breast accounts for only 1-6% of mammary cancer in Caucasian women and is characterized by a poor prognosis; distant metastases frequently appear in fact in an early stage of disease and moreover metastatic spreading follows unpredictable ways. In this study we report on a case of a female patient in whom persistent signs of increased intracranial pressure, following the diagnosis of inflammatory carcinoma of the breast, have been referable to the tumour seeding the meninges in the absence of systemic disease. This peculiar and unusual form of neoplasia is up today a challenge for the clinician, both because of therapeutic difficulty and of unexpected metastases which, in turn, worsen the prognosis. Particularly, in our opinion, meningeal localization must be suspected even in the absence of distant metastases. PMID- 7478077 TI - [Botulinum poisoning in adults: a case report and differential diagnosis]. AB - The authors reports the case of a patient with quadriplegia associated with respiratory failure. A few days before, a paralysis of the oculomotor nerves appeared, which rapidly spreads in cranio-caudal fashion. The occurrence of a similar symptomatology in a patients' relative addressed the diagnosis toward a possible Clostridium botulinum intoxication. Other possible differential diagnoses are reviewed and discussed. PMID- 7478078 TI - [Neuroleptic malignant syndrome. A case report]. AB - A lethal case of neuroleptic malignant syndrome in a 42-year-old woman with a history of bipolar psychiatric disorder under treatment with haloperidol is reported. The patient, hospitalized many times in the past for psychiatric treatment, was then admitted for treatment of a relapse of the disease during a exceedingly hot period. The patient complained of hallucinations and stomach ache. It was necessary to increase the dose of haloperidol to 2 mg, 3 times a day, and to give a single dose of perphenazine enanthate 100 mg. All tests proved normal except for CPK 274 U/l and urea 14 mg/dl. On hospital day 2, went into coma with high temperature, diaphoresis, polyuria, leucocytosis (WBC count 15,440 U/mm3), urea 7 mg/dl, LDH 425 U/l, Na 114 mEq/l, K 2.5 mEq/l. The rapid improvement following hypertonic saline treatment encouraged a diagnosis of water intoxication. On hospital day 7, continuing with the haloperidol treatment, the patient developed a high temperature and deterioration torpor. On hospital day 9, the patient went back into coma with temperature over 40 degrees C (104 degrees F), with fine myoclonia most visible on the face, and muscular rigidity. The CPK, rose from 5,169 to 28,060 U/l in less than 24 hours; the serum myoglobin rose to 41,000 ug/l. On day hospital 11 developed renal, cardiac and respiratory insufficiency. The picture deteriorated and dantrolene was no longer of use in controlling the fever. Then with a fever of over 42 degrees (108 degrees F) and the CPK at 50,000 the patient died. The autopsy demonstrated widespread rhabdomyolysis, a picture of widespread aspecific shock in all organs and picture of myoglobin-induced tabular necrosis of the kidney. PMID- 7478079 TI - [Aortic dissection similar to giant cell arteritis: diagnostic difficulties and efficacy of steroid therapy]. AB - The authors report a case of an acute aortic dissection in a sixty year old patient who two months previously had an ischemic cerebral vascular accident. On the basis of a remittent fever and of raised acute phase proteins the authors suspected a giant cell arteritis as possible pathogenic cause of the clinical presentation. Ruled out infective and neoplastic disorders, after an unhelpful temporal artery biopsy, steroid treatment was introduced at the recommended dosage. Three months after, while reducing steroid therapy, the acute phase proteins raised again to come back to normal values only after restoring full steroid dosage. This pattern of response to steroid treatment may further support the diagnosis of giant cell arteritis even after an unhelpful temporal artery biopsy. PMID- 7478080 TI - [Hysteroscopy in septate uterus. Diagnostic and therapeutic features]. AB - Personal experience of diagnostic and operative hysteroscopy of septate uterus is reported. The diagnosis of septate uterus is an integrated diagnosis that makes use of hysterosalpingography, echography, hysteroscopy and laparoscopy. Of 545 hysteroscopies carried out between January and December 1993, 29 evidenced uterine septum, complete in 6 cases, incomplete in the remaining 23. Of the 29 patients, 7 underwent resection of the septum by the hysteroscopic route. No intra or postoperative complications worthy of note were observed. At follow-up after one month, the uterine cavity in 5 patients proved completely regular while in the remaining 2 one case of fundic fibrous synechia and one of small persistent fibrous bridge, of little importance within the context of the cavity, were observed. The purpose of the present paper is to demonstrate the twofold role of hysteroscopy with regard to the septate uterus; is that hysteroscopy is a diagnostic and a precise operative technique that is atraumatic, fast and of low cost. PMID- 7478081 TI - [Usefulness of hysteroscopy in the management of abnormal uterine bleeding and intrauterine benign disease]. AB - The hysteroscopy is a useful methodology for the diagnosis of uterine intracavitary disorders in women suffering from abnormal uterine bleeding and infertility. The authors report 84 cases who underwent a diagnostic hysteroscopy at the Department of Obstetrics and Gynecology of Ivrea Hospital and then they will discuss the results. In particular, from their study it comes out that the methodology correlated with the histologic results has a sensibility of 100% and a specificity of 85%. PMID- 7478082 TI - [Role of microcolpohysteroscopy at an ambulatory center of level II preventive gynecologic oncology in the study of pre-neoplastic lesions of the uterine cervix]. AB - The microcolpohysteroscopy has proved to be a very reliable methodology for the study of preneoplastic diseases of the uterine cervix especially when located in the endocervix. The authors report 100 cases with pathological Pap tests who underwent the microcolpohysteroscopy. PMID- 7478083 TI - [Validity of instrumental tests, individual and combined, in the rapid diagnosis of extra-uterine pregnancy]. AB - Extrauterine pregnancy is an increasingly frequent pathology. It is also responsible for a high percentage of maternal deaths. This figure is closely correlated to the rapidity of diagnosis. This retrospective study analyses cases of extrauterine pregnancy observed over a 10-year period and aims to evaluate the value, in terms of an early or rapid diagnosis, of various instrumental tests, the used either alone or in association. PMID- 7478084 TI - [Long-term assessment of bone loss in patients treated with GnRH agonists]. AB - GnRH Analogues therapy of estrogen-dependent gynaecological diseases aims at suppression of physiologic ciclic ovaric function producing a hypogonadotropic condition. The first consequence of GnRH Analogues administration is hypoestrogenism; this condition permits the disease regression but, on the other, it causes a negative impact on bone metabolism particularly for prolonged therapeutic schemes (6 months). This study has evaluated the faculty of bone mass protection combining GnRH Analogues therapy with Ipriflavone that stimulates, both "in vivo" and "in vitro", Osteoblastic cells activity. The result of this study showed a significant bone loss in patients treated with GnRH Analogues only. The Ipriflavone association prevented bone loss. PMID- 7478085 TI - [Effect of menstrual cycle on the immune response to surgical stress]. AB - The Authors have studied the effects of different surgical interventions on some factors of humoral and cell-mediated immuno-competence: 112 premenopausal women, operated on mainly for benign conditions, have been considered in order to evaluate the influence of various phases of menstrual cycle on surgical immunosuppression. Two schemes were used to stratify the study sample: in the first analysis, we have subdivided the patients in 59 cases in perimenstrual phase (0 through 6 and 21 through 36 days from last menstrual period, LMP) and 53 in periovulatory (7-20 days from LMP). The second stratification relied on the hormone-dependent phases determined by the putative time of ovulation 14 days after LMP: 51 cases in follicular phase and 61 cases in luteal phase have been considered. Immune parameters have been evaluated before surgery and the day after intervention. The first comparison stresses the worst outcome of immune parameters in patients operated on in the perimenstrual period (CD4, activated lymphocytes and C3 have significantly reduced post-operatively). The second part of the study shows no significant difference in the two groups as to such parametres. PMID- 7478086 TI - [Pharmacologic induction of fetal immobilization for prenatal diagnostic therapeutic procedures]. AB - Neuro-muscular blocking agents are currently administered to the fetus during intrauterine procedures, in order to freeze fetal movements and to avoid traumatic effects. The authors have evaluated three drug regimens (Pancuronium, Vecuronium, Atracurium), both via the intramuscular and the intravascular route of administration to the fetus. The time lapse between injection and disappearance of movements and the duration of paralysis have shown no significant differences for each group. The duration of fetal immobilization, on the other hand, has resulted excessively long for the necessity of the procedure. The authors speculate that immaturity of fetal metabolism can be responsible for the prolonged action of the drug. No side effect related to paralysis have been recorded at birth and after two years follow-up. PMID- 7478087 TI - [Usefulness of a second-level centralized ambulatory service for the staging of diseases of the endometrium and the lower genital tract in a provincial hospital]. AB - In 1991, at the Department of Obstetrics and Gynecology of Ivrea Hospital Hysteroscopy-Microcolposcopy-Vulvoscopy center was set up in order to improve the management of the intracavitary and lower genital tract pathologies. In such center skilled specialists having attended microcolpohysteroscopy and vulvoscopy training courses in specialised institutes are employed. They continuously keep in contact with such institutes for the updating of diagnostic and therapeutic protocols. The authors present their opinions about the usefulness of this center and the results achieved. PMID- 7478088 TI - [Hypertension in pregnancy: management protocols and pregnancy outcome followed at our ambulatory service]. AB - OBJECTIVE: To verify the usefulness of an outpatient Clinic for Hypertension in Pregnancy after 10 years of activity. STUDY DESIGN: A retrospective analysis of the outcomes of pregnancies followed at our outpatient Clinic from 1980 through 1990 was performed. PATIENTS: 607 pregnant women were followed-up: 179 patients developed gestational hypertension during one or more of their previous pregnancies, 275 were affected by chronic hypertension and 213 developed hypertension during the present pregnancy. INTERVENTIONS: Detailed protocols of treatment and management of patients were observed. RELIEFS: Pregnancy outcomes regarding the incidence of preeclampsia and small for gestational age (SGA) newborns was evaluated. The outcome was considered good when gestational age at delivery was more than 36 weeks and neonatal birthweight was adequate for gestational age. RESULTS: The patients with preeclampsia in previous pregnancies developed this complication in 5%; patients with gestational hypertension in the actual pregnancy or with chronic hypertension had a significant proteinuria in 12% and 6.2% of cases respectively. Higher incidence of SGA was found in patients who developed preeclampsia (58%) and in chronic hypertensive pregnancies (34%), while only 13% of SGA was in patients with hypertension in previous pregnancies. CONCLUSION: From our experience we can conclude that an outpatient Clinic for hypertension in pregnancy is extremely useful to give the opportunity for application of same criteria of management and therapy. This allow to admit to the hospital only patients with any signs of development of preeclampsia. PMID- 7478089 TI - [Autotransfusion in gynecologic surgery. Analysis of an experience]. AB - The autologous program is the most reliable method used in avoiding the consequences of homologous blood transfusion (particularly hepatitis and HIV) for patients who must undergo surgery. The autologous transfusion program's prerequisites are less restrictive than those concerning the homologous transfusion program. There are different modes to accomplish the autologous blood transfusion program according to the chosen technique. The risks involved in autologous blood donation are the same as these involved in a normal blood donation and they are not considered a restrictive factor. The practice of the autologous program in Gyn surgery is not different from what has been done in general surgery. Finally, shown below, is a study of the autologous transfusion experience in Gyn surgery performed in the OB/GYN service at San Daniele del Friuli Hospital, Udine for the period 1989-1993. A new method of autologous donation has been achieved in this ward since 1987. PMID- 7478090 TI - [Experience with autologous transfusion in obstetrics]. AB - The consequences of homologous blood transfusion have a remarkable impact on the obstetric population which mainly consists of young healthy women. The risk involved in possible post-transfusion complications is directly proportional to the number of homologous blood units that are being transfused. Autologous transfusion is riskless for the fetus' well being. The main obstetrical problem is that it is impossible to foresee to the maximum extent all cases in need of blood transfusion which might get either an excessive amount or an insufficient amount of autologous transfusion. Blood predonation is the most suitable method in the obstetrics service. Reported below are the findings concerning the autologous transfusion experience performed by the OB/GYN Department at the Hospital San Daniele del Friuli, Udine for the period 1989-1993, where a new method of autologous donation was performed on 44 pregnant women who delivered by C-section. The autologous program is considered an easy one in the OB/GYN field, without complications for the fetus' well being and also feasible for small hospitals. PMID- 7478091 TI - [Malignant gynecologic neoplasms during menopause. Authors' experience]. AB - From January 1991 to December 1993 409 post-menopausal women were observed at the Center for the Study of Climateric Disorders of the Department of Obstetrics and Gynecology of the University of L'Aquila. The aim of the study was to evaluate the incidence of gynecologic neoplasm among this group of women. A total of 48 women at risk for malignant tumors were detected. The relationship between the climateric age and the oncologic risk is discussed. PMID- 7478092 TI - [The role of echography in the management of fetal macrosomia]. AB - Macrosomia remains an important cause of perinatal morbidity and mortality and maternal morbidity, which arise mainly from birth injury and asphyxia and increased rate or cesarean section respectively. In spite of many attempts to identify the patients at risk for fetal macrosomia, the perfect prediction has not yet emerged. In the last years, many papers have reported methods for ultrasonographic determination of fetal weight. It should be emphasized that most of these reports have cautioned that sonographic predictive accuracy declines as fetal weight increases toward 4000 g. In our study, sonographic prediction of diagnosis of macrosomia was performed in 73% of cases. In addiction, in case with a correct sonographic diagnosis of macrosomia, a higher incidence of primary operative delivery was reported, and when operative delivery for failure to progress was performed, the time of labor was shorter in patients with ultrasound diagnosis of macrosomia. The proportion of babies with apgar score greater than 7 at 1 minute, the incidence of shoulder dystocia and birth trauma was similar in the patients with and without prenatal ultrasound diagnosis of macrosomia. This study does not justify the use of elective cesarean section as a method of preventing persistent infant morbidity. An accurate antenatal diagnosis of macrosomia with ultrasound has been shown to offer an important usefull in the clinical management of labor and delivery. PMID- 7478093 TI - [The role of hysteroscopy in the follow up of post-mastectomy patients undergoing adjuvant therapy with Tamoxifen]. AB - The authors report on up to date knowledge of the risk of endometrial carcinoma women operated on mastectomy for breast carcinoma and treated with TAM. Starting from their own clinical and scientific experience, the authors follow a group of such patients with a strict monitoring, to ascertain the eventual comparison of dysplastic and neoplastic endometrial pathologies. The group numbers 18 patients and the aim of the study is to evaluate the importance of hysteroscopy as a diagnostic approach for this iatrogenic pathology. The authors affirm the validity of this partially invasive diagnostic method that has to be integrated with clinical and laboratory parameters that are justified by the cost/benefit ratio always favourable for the diagnosis of a neoplastic pathology. PMID- 7478094 TI - [Thymopentin and cervico-vaginal HPV infection associated with CIN]. AB - 46 patients affected by HPV+CIN were treated with Thymopentin i.m. HPV disappeared in all cases. We also observed complete disappearance or only a partial remission of CIN in 82.6% of cases. PMID- 7478095 TI - [Evaluation of the effect of a "Mediterranean diet" and pre-pregnancy body mass on fetal growth]. AB - In this study we compared the pregravidic body mass index (Quetelet index) with some variables recorded during pregnancy (caloric intake; weight increase, neonatal birth weight) in a group of patients consumers of a typical mediterranean diet. We studied the dietary habits of 100 pregnant women with gestational ages between 13 and 38 weeks. Each woman has been provided with daily, for seven consecutive days. The data have been analyzed evaluating, for each dietary componene, the caloric, proteic, glucidic, lipidic and microelements share. Our results showed degree of pregravidic obesity. However also in overweight patients that used diets with caloric intake as low as about 1600 Kcal/day, the neonatal weights were always within the normal range, confirming the adequacy of the "mediterranean diet" to the specific nutritional needs in pregnancy. Neonatal weight was better correlated to the pregravidic IQ than to the caloric intake. PMID- 7478096 TI - [Cesarean section and spontaneous birth: clinical aspects and maternal psychodynamic impact]. AB - Cesarean delivery was considered a medical saving-operation for a very long time. Few editorials focus on psychological results of cesarean deliver in respect of the mother, the relationship mother-newborn, the Hospital, and family environment. The authors examined psychodynamic effects of cesarean and vaginally deliver puerperas, comparing different experiences of attainment of maternity. An interview was administered to 39 cesarean and 41 vaginally delivered mothers, 4-6 days after birth. All these women were primiparas and similar for social abd educational level, marital status, employment rate, age and gestational age of the birth. Women with malformed babies and obstetric pathologies were excluded. After elaborating these data we can understand that there are many factors interfering with maternal gratification in the era of cesarean delivery. Results underline the main consequences of surgical delivery, focus on the privation of creative onnipotence of vaginally birth. This is reflected on quality of the first contact with the newborn. Therefore, for the medical staff, it is important to reduce the troubles caused by the anaesthesia, the lack of "the first contact" mother-newborn, taking care about puerperas" psychosomatic problems. PMID- 7478097 TI - [Terminology and classification problems in relation to vulvar pathology]. AB - The aim of this paper is to update the physicians (gynecologists, dermatologists and pathologists) on the evolution of vulvar disease terminologies. In doing that the authors illustrate briefly the fundamental steps which led to present classifications of the International Society for the Study of Vulvar Disease (ISSVD). The classification of "non neoplastic epithelial disorders" together with that of "intraepithelial alterations" are illustrated and compared with the terminologies previously employed. The last ISSVD definition of "superficially invasive carcinoma" of the vulva is also presented and discussed. The authors concluded that even if all these ISSVD classifications represent an important effort for reaching a common language for a better international exchange of different experiences, nevertheless an improvement of these terminologies is still requested. PMID- 7478098 TI - [Parkinson syndrome induced by veralipride]. AB - Veralipride, an antidopaminergic drug commonly prescribed to counteract postmenopausal symptoms, may cause reversible parkinsonism. A 49-year-old healthy woman reported a 5 month history of progressive slowing of voluntary movements, postural changes, e.g. sit-to-stand manouevres, and gait. Clinical examination revealed moderate hypokinesia, hypomimia and plastic rigidity of the four limbs. Locomotor oscillations of the upper limbs were decreased. She had been taking 100 mg of veralipride daily to counteract postmenopausal symptoms (vasomotor flushes and irritability) for 17 months uninterruptedly. The drug was withdrawn. The patient's clinical picture fully normalized within 20 days. One year later she reported to be still asymptomatic. This seems to be a case of parkinsonism induced by veralipride, a drug known to cause other extrapiramidal signs such as bucco-facial or limb dyskinesia. To our knowledge, only one other such case has been published (a 77-year-old French woman who had been taking veralipride, prazepam and nicergolin). In both cases, the drug had been administered for longer and without intervals. In our patient the motor disturbances could not have been associated with either advanced age or interaction with concurrent medications. Therefore, this case is suspected for veralipride-associated parkinsonism. In conclusion, intoxication should be thought in case of parkinsonism arising in women taking this drug. PMID- 7478099 TI - [Malignant vulvo-vaginal melanoma. Review of the literature and a clinical case]. AB - Vaginal melanoma is a quite rare pathology and represent the 2.6% of overall primitive vaginal tumors. The development of this disease is very aggressive and the 5 years survival is only the 7%. A radical surgical effort seems not improve the survival than a wide local exision. The authors give in this article a wide review of the literature about the vaginal melanoma and the personal observation. We believe that despite the unfrequency of this disease, it could be always taken in mind on the clinical approach with aged women. PMID- 7478100 TI - [Peroperative diagnosis of pure retroperitoneal neurinoma misdiagnosed as uterine leiomyoma]. AB - A case of pure neurinoma of the retroperitoneal space developing in the pelvis is described. This tumor (phi max, 15 cm) responsible for intense compressive symptomatology involving the pelvic organs, was at first mistaken for a uterine fibroid. The ultimate diagnosis both of neurinoma as well as every other neoplasm originating in the retroperitoneum can sometimes be very hard to obtain. Those tumors can indeed be misdiagnosed for other more common conditions both clinically and instrumentally and the rarity of this particular localization adds further trouble to the matter. PMID- 7478101 TI - How to improve your manuscript (or how to increase your chances of manuscript acceptance) advice from an editor. PMID- 7478102 TI - The inscrutable renal cyst in ADPKD. PMID- 7478103 TI - Hyperglycaemia--pathophysiological aspects at the cellular level. PMID- 7478104 TI - Mycophenolate mofetil (RS 61443): nothing new under the sun or an important break through in the field of transplantation? PMID- 7478105 TI - Does CAPD provide adequate dialysis? PMID- 7478106 TI - Renal disease--another effect of silica exposure? PMID- 7478107 TI - Treatment of IgA nephropathy. PMID- 7478108 TI - Abnormal glycosylation of IgA: is it related to the pathogenesis of IgA nephropathy? PMID- 7478109 TI - Peritoneal aspergillosis--pitfalls in the diagnosis of a rare disease. PMID- 7478110 TI - Chronic rejection and hypertension: a chicken-and-egg problem. PMID- 7478111 TI - When dialysis becomes worse than death. PMID- 7478112 TI - Innovative strategies for pharmacological intervention in immune damage to the kidney. PMID- 7478113 TI - A tribute to Eric Muirhead. Evolution of the Medullipin concept of blood pressure control. PMID- 7478114 TI - Discontinuation of treatment among Italian diabetic patients treated by renal replacement therapy. AB - Death from dialysis termination has been extensively surveyed in Canada, the United States, and Australia. In the US old age and the presence of diabetes has been associated with treatment withdrawal. On the other hand, information for Europe is very scarce. We addressed the issue of dialysis termination in Italy in both a cohort of diabetic patients starting RRT in 1987, and two age-, sex-, type of RRT, and unit-matched cohorts of diabetic and non-diabetic patients alive on RRT treatment on 31 December 1987. Follow-up was available till 31 December 1991. Dialysis termination accounted for 1.1% of the known causes of death in the incident diabetic cohort and for only 0.5% and 0.9% of the prevalent diabetic and non-diabetic cohorts respectively. In Italy, diabetes is not associated with higher rates of dialysis termination and this cause of death seems uncommon among the overall Italian RRT population. We cannot, however, exclude a predialysis selection against patients presenting with an old age or comorbid conditions. PMID- 7478115 TI - Association of M235T variant of the angiotensinogen gene with familial hypertension of early onset. AB - A higher frequency of a variant of the angiotensinogen gene characterized by a transition in exon 2 causing a replacement of methionine by threonine (M235T) has recently been found in hypertensive individuals, but not all authors were able to confirm this observation. We examined (i) 219 patients with primary hypertension, (ii) 92 normotensive controls (spouses), and (iii) a sample of the general population (blood donors, n = 139). Analysis of genomic DNA was performed by PCR amplification and alleles were separated on agarose gels. In the general population and in normotensive spouses the respective frequencies of the T and M alleles were: general population: M = 0.6, T = 0.4; normotensive spouses: M = 0.59, T = 0.41. A significantly higher frequency of the 235T allele was found in hypertensive individuals with a family history of hypertension and an onset of hypertension before 50 years of age (spouses: 0.41 versus HT with age of onset < or = 50 years and family history of HT: 0.56; P = 0.01 by chi 2). In conclusion, the present study confirms the observation of a higher frequency of the 235T allele of the angiotensinogen gene in hypertension and identifies individuals with family history and early onset of hypertension as individuals at risk. PMID- 7478116 TI - Cyclosporin A reduces albuminuria in experimental anti-GBM nephritis independently from changes in GFR. AB - Cyclosporin A (CsA) can reduce proteinuria in various forms of human and experimental glomerulonephritis. This antiproteinuric effect of CsA may be the consequence of diminution of immunological damage, a drug-induced decrease of GFR, or changes in permselectivity of the glomerular basement membrane (GBM). We studied the antiproteinuric effect of CsA in the heterologous phase of a passively induced anti-GBM nephritis in the mouse. This passive model is characterized by acute exudative glomerular lesions and a dose-dependent albuminuria. Rabbit anti-mouse GBM antibodies were administered intravenously in C57B110 mice at day 4, after 3 days of pretreatment with either CsA (75 mg/kg body weight) (n = 15) or olive oil (OO, controls, n = 15) orally. CsA did not influence the severity of the histological lesions. Albuminuria was substantially reduced by CsA (CsA 1.6 +/- 1.8; OO 5.6 +/- 3.2 mg/18 h; P < 0.002). There was a considerable concomitant reduction of the GFR by CsA, as measured with a 51Cr EDTA single-shot plasma clearance technique before (day-1) and during treatment (day 4): GFR ratio day 4/day-1 for CsA, 0.4 +/- 0.1; for OO controls, 1.1 +/- 0.6; P < 0.01. This drug-induced decrease of GFR was prevented by simultaneous treatment with phenoxybenzamine (PB) twice daily 45 micrograms orally for 4 days (GFR ratio day 4/day-1 for PB and CsA, 0.9 +/- 0.4; controls (PB and OO), 1.0 +/- 0.4; P = NS). Although the CsA-induced GFR reduction was prevented, CsA still reduced albuminuria significantly (PB and CsA, 2.2 +/- 1.8; controls (PB and OO), 5.6 +/- 1.8 mg/18 h; P = 0.003).(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7478118 TI - Wegener granulomatosis is associated to exposure to silicon compounds: a case control study. AB - Wegener granulomatosis is a rare disease of unknown aetiology. In the majority of these patients the kidney is involved in the disease process. We performed a case control study to evaluate the role of occupational exposure in the development of Wegener granulomatosis with renal involvement. The occupational histories of 16 cases with clearly established diagnosis of Wegener granulomatosis with renal involvement were compared with those of 32 age- and sex-matched controls. It was observed that inhalation of silicon-containing compounds such as silica and grain dust gave a nearly sevenfold risk for Wegener granulomatosis. Further epidemiological and experimental work needs to be performed in order to corroborate these findings. PMID- 7478117 TI - Beta 1 and beta 3 integrin upregulation in rapidly progressive glomerulonephritis. AB - The expression and distribution pattern of beta 1 (alpha 1-alpha 6) and alpha v beta 3 integrins and ICAM-1 and VCAM-1 counter receptors were evaluated by an immunohistochemical technique on eight renal samples from patients affected by rapidly progressive glomerulonephritis (RPGN) of different aetiologies. In all cases integrins and counterreceptors displayed similar patterns. On tubular cells of renal cortex, a marked upregulation of alpha 2 beta 1, alpha 3 beta 1, alpha 5 beta 1, alpha v beta 3 integrins and VCAM-1 was observed with as many as 60-90% of tubular cross-sections labelled, while a strong ICAM-1 reactivity was limited to the luminal surface. The same adhesion molecules were also uniformly expressed on crescentic cells. In glomeruli, integrin upregulation occurred only on apparently preserved capillary tufts, i.e. in an early stage of lesion, while collapsed and sclerotic tufts showed a reduced integrin expression. In addition a morphometric study of extracellular matrix (EM) proteins cellular fibronectin and tenascin showed a 9.56 +/- 1.9-fold and 3.35 +/- 0.6-fold increase respectively in these proteins, as compared to normal kidney (P < 0.001). The upregulation of alpha v beta 3 on podocytes might play a role in the adhesion of crescentic cells. An increased production of cytokines, in particular transforming growth factor-beta, might induce augmented deposition of EM proteins and upregulation of beta 1 and beta 3 integrins in RPGN. PMID- 7478119 TI - The pathogenetic significance of C5b-9 in IgA nephropathy. AB - Renal biopsies from 20 patients with IgAN were retrospectively studied using monoclonal antibodies against T cells, monocytes/macrophages (MM), HLA-DR-related antigen and the C5b-9 neoantigen. Glomerular C5b-9 deposits were mainly found in the mesangial areas and showed an association with IgA (P < 0.005) and C3 deposits (P < 0.001). Interstitial T cells and MM were highly correlated with the interstitial DR + ve cells (P < 0.001 and P < 0.0005 respectively). Tubular C5b-9 deposition was observed on the tubular basement membranes and related to the numbers of interstitial T cells (P < 0.005), MM (P < 0.005) and DR + ve cells (P < 0.01) as well as to the tubular DR expression (P < 0.025). The severity of tubular atrophy and interstitial fibrosis showed a positive correlation with the interstitial T cells, MM and DR + ve cells, as well as with the intensity of tubular C5b-9 deposition (P < 0.05 and P < 0.05 respectively). Plasma creatinine on presentation was correlated with the numbers of interstitial T cells (P < 0.05), MM (P < 0.01), interstitial DR + ve cells (P < 0.005), and tubular C5b-9 deposits (P < 0.005). No correlation between glomerular T cells, MM, and C5b-9 deposits with plasma creatinine was seen. During follow-up, renal function deteriorated in those patients with the more extensive tubular C5b-9 deposits. In conclusion, glomerular C5b-9 deposition seems to be secondary to the IgA and C3 deposition. In contrast, tubular C5b-9 is related to the numbers of interstitial T cells and MM. Activated interstitial mononuclear cells may lead to the tubular deposition of C5b-9, which eventually might contribute to the development of tubulointerstitial lesions (TIL) and renal function impairment. PMID- 7478120 TI - Can immunosuppressive drugs slow the progression of IgA nephropathy? AB - The role of immunosuppressive drugs in the treatment of IgA nephropathy (IgAN) remains controversial. The effect of treatment with prednisolone and azathioprine on the clinical course of patients with IgA nephropathy is described in this retrospective study. One hundred and fourteen patients, 66 treated (age 13-77 years) and 48 untreated (age 15-64 years), were evaluated. The two groups of patients differed significantly with respect to heavier proteinuria (median 3.6 g/day, range 0.2-18 g/day), lower serum albumin level (< 40 g/l) and more severe renal histopathological involvement in the treated group (P < 0.01). Oral prednisolone 40 mg/day and azathioprine 2 mg/kg BW/day was commenced initially and after gradual tapering was continued at low dose (5 mg/day) for a median duration of 24 months (range 12-98). The median duration of follow-up was 46 months (range 12-180). The clinical course was defined as progressive or non progressive on the basis of serial serum creatinine (Scr). Of the patients who presented with renal impairment (Scr > 110 mumol/l), a non-progressive course was observed in 79.5% patients of the treated group (n = 39), while only in 36% of the untreated group (n = 22), the difference was statistically significant (P < 0.001). Slopes of reciprocal of Scr versus time were also calculated by linear regression analysis to represent the trend of renal function for patients who had had 3 or more years follow-up (n = 101). An analysis of variance of these trends in patients with renal impairment at presentation (n = 51) showed significant recovery of renal function in the treated group (n = 33) and a decline of renal function in the untreated group (n = 18, P = 0.004). There was no significant effect of the treatment on proteinuria. The histopathological features that favoured response to the treatment were mesangial proliferation, capsular adhesions and interstitial infiltration on light-microscopy, C3 and fibrin deposits on immunofluorescence (P < 0.05).(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7478121 TI - Late outcome of a controlled trial of enalapril treatment in progressive chronic renal failure. Hard end-points and influence of proteinuria. AB - An earlier controlled trial showed that over an average of 26 months, enalapril slowed the progression of chronic renal failure. Following completion of the trial, the patients continued to receive antihypertensive treatment according to ordinary clinical criteria. All but four patients in the enalapril group remained on that drug, and two patients in the control group were switched to an angiotensin-converting enzyme (ACE) inhibitor. In the present study the fate of the 70 patients 44 months after termination of the trial was investigated, with a total follow-up of around 7 years. In the original enalapril group, 12 of the 35 patients (34%) were alive without renal replacement therapy versus five of the 35 patients (14%) in the control group. This difference of 20% in favour of having been in the enalapril group in the original trial was significant (P = 0.05; 95% confidence limits 0.5-39.5%). The influence of baseline proteinuria on clinical outcome was analysed. In the original control group, baseline renal clearances of albumin (Calb) and immunoglobulin G (CIgG) were significantly lower in patients surviving without renal replacement therapy at follow-up than in patients who ultimately developed end-stage renal failure (ESRF) (P < 0.05). In the original enalapril group, these baseline clearances were equal in the two renal outcome groups. In all patients, baseline Calb and CIgG were negatively correlated with the rate of change in GFR during the controlled trial (r = -0.37, P < 0.01 and r = -0.28, P < 0.05).(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7478123 TI - Pneumocystis carinii pneumonia in renal transplant recipients. AB - In 1991 and 1992 Pneumocystis carinii pneumonia (PCP) was diagnosed in 28 renal transplant recipients. The incidence of PCP in our renal transplant centre was remarkably increased from 1.1% before 1991 to 11.5% in 1991-1992. We compared 28 PCP patients with a control group of 27 renal transplant recipients, matched for transplantation day and without an episode of PCP. The mean age was significantly higher in the PCP group (50 +/- 13 versus 38 +/- 13 years). We observed no differences in basic immunosuppressive and rejection treatment nor in antibiotic consumption, number of hospitalization days, and incidence of CMV infection. In March 1993 we introduced PCP prophylaxis. More than 140 renal transplant recipients received co-trimoxazole, starting 1 day after transplantation and continued for a period of 4 months. To the time of writing no one in this group had developed PCP. PMID- 7478122 TI - The renal manifestations and outcome of thrombotic thrombocytopenic purpura/hemolytic uremic syndrome in adults. AB - Thrombotic thrombocytopenic purpura/Hemolytic uremic syndrome (TTP/HUS) is generally regarded to be a rare disease. The present study was undertaken to identify presenting features, prognostic variables, pathological features and outcome associated with TTP/HUS. The present study is a retrospective chart review of 68 patients treated with plasmapheresis for TTP/HUS at a single tertiary referral medical institution from 1980-1992. The annual number of patients with TTP/HUS treated with plasmapheresis increased from an average of one case per year in 1980 to nine cases per a year in 1992. The in-hospital mortality for patients presenting with TTP/HUS was 25%. Forty four percent of patients presented with an elevated serum creatinine, and 16% required hemodialysis support. Of the seven patients who survived and required hemodialysis support only two patients continued on dialysis. None of the patients presenting with a normal serum creatinine required dialysis at any time in their course. Patient age, sex, presenting platelet count, white blood cell count, hemoglobin level and presence of neurological disease were not significantly associated with death or need for dialysis. The histopathological features of TTP/HUS (fibrin/platelet thrombi in renal vessels and glomeruli, fibrinoid necrosis of vessel walls) were found in all five cases autopsied. The incidence of TTP/HUS may be increasing. Alternative possibilities for the increased frequency of cases seen include greater diagnostic suspicion and referral bias. Despite the use of plasmapheresis, mortality during the initial hospital admission was almost 25%. In retrospect prognosis could not be predicted based on admission biochemical or clinical variables. The majority of patients who developed acute renal failure and survived to hospital discharge recovered renal function and became independent of dialysis. PMID- 7478124 TI - Diagnosis of cytomegalovirus (CMV) disease in renal allograft recipients: the role of semiquantitative polymerase chain reaction (PCR). AB - BACKGROUND: Cytomegalovirus disease remains a significant cause of morbidity and mortality in the renal allograft recipient. There is a need for rapid and sensitive techniques predictive of CMV disease to allow initiation of early antiviral therapy. METHODS: Seventy-seven renal allograft recipients were enrolled in a prospective study where CMV viruria (shell vial culture/DEAFF test), viraemia (shell vial culture), serology and detection of virus DNA in peripheral blood leukocytes by PCR (CMV DNAemia) were correlated with clinical evidence of CMV disease. RESULTS: Serology and shell vial culture had poor sensitivity for the early diagnosis of CMV disease. CMV DNAemia appeared to correlate with active virus replication. CMV DNAemia had a sensitivity and negative predictive value of 100% and a positive predictive value of 27% for CMV disease. Patients with symptomatic CMV disease were shown to have higher levels of CMV DNAemia than those with asymptomatic infection. CONCLUSIONS: A negative CMV DNAemia result excluded CMV disease with confidence, but a positive result (given the low positive predictive value) did not by itself provide a reliable guide for the initiation of pre-emptive antiviral therapy. However, the semiquantitative CMV DNAemia result taken together with the clinical findings provided useful information for patient management. PMID- 7478125 TI - Pre-transplant hypertension: a major risk factor for chronic progressive renal allograft dysfunction? AB - Despite of advances in 1-year survival rates of renal allografts, no comparable achievements have been made in long-term graft survival. To identify risk factors for chronic progressive renal allograft dysfunction we conducted a retrospective study in 639 patients transplanted between 1983 and 1990. Graft function was assessed by the slope of individual 1/creatinine regression lines and chronic progressive graft dysfunction was defined as a slope of the 1/creatinine line of > 0.1 dl/mg/year, indicating a loss of glomerular filtration rate of > 10 ml/min/year regardless of the initial serum creatinine value. A number of possible risk factors were determined and analysed by linear regression analysis. One hundred and six patients (16.6%) showed chronic progressive graft dysfunction. No correlation was found between the rate of functional deterioration and the age and gender of the donor or the recipient, the blood group, the prevalence of hepatitis B or C, the number of blood transfusions, the total ischaemia time, or the number of kidneys from female donors grafted into male recipients. Chronic progressive graft dysfunction was associated with the number of HLA-B/DR mismatches (P = 0.04) and with a first acute rejection episode later than 60 days after transplantation (P < 0.001). Chronic progressive graft dysfunction also occurred in the absence of an acute rejection episode. Significantly (P < 0.001) more patients with chronic progressive graft dysfunction were hypertensive not only 12 months after transplantation, but also at the time of transplantation, indicating that hypertension may not only be secondary to deteriorating graft function, but that hypertension per se leads to graft damage and initiates chronic progressive graft dysfunction. All efforts should be made to control blood pressure adequately to improve long-term survival of renal allografts. PMID- 7478127 TI - Effect of having a functioning cadaveric renal transplant on cardiovascular mortality risk in patients on renal replacement therapy. AB - Our aim was to estimate the effect of having a functioning cadaveric renal transplant on the risk of cardiovascular and total mortality in patients on renal replacement therapy (RRT). We retrospectively studied (by the Cox proportional hazards regression model) 195 subjects who began RRT with maintenance dialysis in our centre from 1 January 1978 to 31 December 1991. Out of this number, 76 patients received a cadaveric kidney transplant. Cardiovascular abnormalities at the onset of RRT were the principal independent determinant of both total and cardiovascular mortality risk. As compared to patients on dialysis with the same duration of RRT, patients with a functioning renal transplant for more than 1 year had a significantly lower total mortality risk (mean relative risk (RR): 0.48 (0.25-0.91) (95% confidence limits in parentheses), P = 0.03), an effect whose significance disappeared after adjustment for pretreatment conditions (RR = 0.62 (0.30-1.30), P > 0.3). However, the beneficial effect of a functioning renal transplant for more than 1 year on cardiovascular mortality risk was significant, both before (RR = 0.21 (0.06-0.74), P = 0.02) and after the adjustment for pretreatment conditions (RR = 0.32 (0.11-0.90), P = 0.035). During the first year after a successful transplantation the beneficial effect of having a functioning transplant on cardiovascular mortality risk was only weakly attenuated. During RRT a functioning cadaveric renal transplant decreases cardiovascular mortality risk partially independently of the better pretreatment status of the patients selected for transplantation. PMID- 7478126 TI - Effect of interferon-gamma in dialysis fluid on peritoneal defence in rats. AB - BACKGROUND: A major drawback of continuous ambulatory peritoneal dialysis (CAPD) is the occurrence of peritoneal infection. This might be explained by a non optimal phagocytic capacity of peritoneal cells which can be improved by stimulating factors. AIM: To investigate the effect of addition of interferon gamma (IFN) to dialysis fluid with various glucose concentrations or to saline (as control) on the peritoneal defence against Staphylococcus aureus in an experimental dialysis model in rats. METHODS: Twenty-four hours after the administration of either dialysis fluid containing various glucose concentrations or saline with or without IFN, bacteria were injected intraperitoneally. At the time of the bacterial infection and 24 h later cellular and bacterial parameters were studied. RESULTS: The addition of IFN to dialysis fluid or saline resulted in a significant (P < 0.01) increase in the number of peritoneal macrophages at the time of infection; this was accompanied by a significant increase in both the number of Ia-positive peritoneal macrophages (P < 0.01) and the production of nitrite by macrophages (P < 0.05) at the time. IFN in dialysis fluid as well as in saline significantly (P < 0.01) reduced the recovery of bacteria from the peritoneal cavity 24 h after infection. Only the absence of IFN glucose increased the recovery of bacteria from the peritoneal cavity at the same time. CONCLUSION: In this experimental model the addition of IFN to dialysis fluid lowered the recovery of staphylococci from the peritoneal cavity by means of activation of an increased number of macrophages. PMID- 7478128 TI - Revascularization of occluded haemodialysis fistulae with the Hydrolyser thrombectomy catheter: description of the technique and report of six cases. AB - Our purpose was to describe initial experience with a new mechanical thrombectomy catheter for treating haemodialysis fistulae. The Hydrolyser thrombectomy catheter combines thrombus fragmentation by a saline jet with simultaneous low pressure aspiration. This catheter was used in six cases of dialysis-fistula occlusion. In four instances this was a polytetrafluoroethylene loop, in two cases a Brescia-Cimino fistula. In all cases rapid revascularization was obtained without complications. Reocclusion occurred in one patient, immediate reintervention being successful. Our initial results suggest that the Hydrolyser is a quick, minimally invasive, and useful alternative to the presently accepted methods of revascularizing occluded haemodialysis fistulae. PMID- 7478129 TI - Elimination of gadolinium-DTPA by peritoneal dialysis. AB - Magnetic resonance urography (MRU) after i.v. injection of gadolinium-DTPA (0.1 mmol/kg bodyweight) was performed in a patient with transplant dysfunction after renal allograft transplantation. Renal replacement therapy was accomplished by means of peritoneal dialysis. Gadolinium-DTPA concentrations in urine and dialysate were measured repeatedly during a 48-h period with inductively coupled plasma atomic emission spectrometry. The total 24-h creatinine clearance was 10.8 ml/min on day 1 and 13.7 ml/min on day 2. The corresponding total gadolinium clearance was 12.8 ml/min and 15.8 ml/min respectively. Gadolinium-DTPA was removed from the body through peritoneal dialysis at a mean clearance rate of 5.13 ml/min. Plasma half-life of gadolinium-DTPA was prolonged to 9 h. No side effects due to gadolinium-DTPA were noted. MRU provided a better visualization of the morphology of the urinary tract than scintigraphic studies. By measuring the changes of signal intensity, the gadolinium excretion could be calculated. PMID- 7478130 TI - Henoch-Schonlein disease with IgA nephropathy associated with chronic alcoholic liver disease. PMID- 7478131 TI - Disseminated cutaneous infection due to Mycobacterium chelonae in a patient with rheumatoid arthritis, amyloidosis, and renal failure. PMID- 7478132 TI - Occult adrenal insufficiency secondary to amyloidosis in the context of chronic renal failure. PMID- 7478133 TI - Aspergillus peritonitis in peritoneal dialysis: case report and a review of the literature. AB - Aspergillus peritonitis is a rare complication of continuous ambulatory peritoneal dialysis. The case is described of a 68-year-old man in whom Aspergillus fumigatus was isolated from the peritoneal dialysate after recurrent peritonitis with Gram-negative rods in association with diverticulosis. Treatment consisting of removal of the catheter and intravenous administration of amphotericin B followed by oral itraconazole was successful. A review of the sparse literature (12 cases) displays uncertainties regarding diagnostic awareness, culture diagnosis, and therapeutic management. Next to institution of appropriate antifungal therapy, early removal of the peritoneal dialysis catheter is recommended, as delayed removal of the catheter appears to be associated with increased mortality and morbidity. PMID- 7478134 TI - The description of essential hypertension by Frederick Akhbar Mahomed. PMID- 7478135 TI - A febrile patient with a renal space-occupying lesion evolving into renal abscess: acute focal bacterial nephritis. PMID- 7478136 TI - A young man with recurrent stones and renal failure. PMID- 7478137 TI - Influence of ACE inhibition on haemoglobin and haematocrit in essential hypertensive patients. PMID- 7478138 TI - Schonlein-Henoch glomerulonephritis complicating diabetic glomerulosclerosis. PMID- 7478139 TI - Successful treatment of acute renal failure related to bone-marrow transplantation. PMID- 7478140 TI - Tumour markers in chronic renal failure and haemodialysis patients. PMID- 7478141 TI - Glucose metabolism in normoglucaemic haemodialysis patients: a possible role for biotin? PMID- 7478142 TI - Lysis-centrifugation method in peritonitis diagnosis. PMID- 7478143 TI - Regulation of bone cells by 1,25(OH)2D3. PMID- 7478144 TI - The spectrum of renal bone disease. PMID- 7478145 TI - The need to treat predialysis patients. PMID- 7478146 TI - The use of alfacalcidol in the prevention of bone disease in early renal failure. PMID- 7478147 TI - Optimum route of administration of vitamin D in renal failure. AB - Vitamin D preparations [1-alpha-hydroxycholecalcifol (1-alpha OHD3) or 1,25 dihydroxycholecalciferol (1,25(OH)2D3)] may be administered orally, subcutaneously, intraperitoneally or intravenously. They may be given daily, sometimes in divided doses, or intermittently in large bolus doses, usually three times per week. A further variable is the timing of administration, which may be at night when the gut calcium load is at a minimum. In at least some studies high dose intermittent bolus administration of vitamin D can reduce parathyroid hormone (PTH) secretion by a mechanism separated in time from an increase in ionized calcium (iCa2+). In addition, some but not all studies have shown an improvement in calcium set point and in parathyroid gland sensitivity to calcium. It is also clear from a number of studies that this treatment can succeed where conventional daily administration has failed. Bolus intravenous therapy is most conveniently given after haemodialysis treatment sessions. Bolus oral therapy, perhaps administered prior to sleep, may achieve a similar objective (PTH suppression without hypercalcaemia) in patients on continuous ambulatory peritoneal dialysis (CAPD). It is also becoming apparent that the parathyroid gland may yet again escape following bolus therapy. The success of bolus therapy may lie in several not mutually exclusive mechanisms: high peak concentrations of 1,25(OH)2D3 directly affecting the parathyroid gland, improved compliance, and a small but significant increase in plasma iCa2+. Further long-term controlled trials are needed before bolus therapy can be generally recommended. PMID- 7478148 TI - Genesis of disturbed vitamin D metabolism in renal failure. PMID- 7478149 TI - Optimum mode of administration of vitamin D in renal failure. Insights from experimental data. AB - In summary, the biological effects of 1,25(OH)2D3 on the parathyroid glands probably persist for several days, although the plasma half-life of 1,25(OH)2D3 is only several hours. This is a theoretical argument for 1,25(OH)2D3 pulse therapy, and suggests that spacing of pulses could be chosen even wider (one-two pulses per week) than was previously thought (three pulses per week). Under the conditions of experiment in subtotally nephrectomized rats, the biological effect on the parathyroid glands was correlated best with the achieved peak 1,25(OH)2D3 serum concentration. This would be another argument for pulse therapy with active vitamin D metabolites. PMID- 7478150 TI - Report on management of renal failure in Europe, XXIV, 1993. PMID- 7478151 TI - Cellular correlates of neuronal hyperexcitability in the vicinity of photochemically induced cortical infarcts in rats in vitro. AB - Intrinsic membrane properties and synaptic responses of neocortical neurons located lateral to photochemically induced ischemic lesions were investigated using neocortical slice preparation. In comparison to neurons from control slices, these neurons had a significantly less negative resting membrane potential without any significant change in input resistance. In addition, gamma aminobutyric acid (GABA) mediated synaptic inhibition was found to be less efficient; the conductances of both the early and late inhibitory postsynaptic potentials (IPSPs) were significantly smaller, and the reversal potential of the early IPSP was shifted to a more positive value. In some of the neurons, 'epileptiform' postsynaptic potentials could be elicited, which were abolished after wash-in of the N-methyl-D-aspartic acid (NMDA)-receptor antagonist D-2 amino-5-phosphonovaleric acid (AP-5). The results provide a possible explanation for the hyperexcitability found in the vicinity of cortical infarcts. PMID- 7478153 TI - Amoeboid microglial response following X-ray-induced apoptosis in the neonatal rat brain. AB - The phagocytic response following X-ray-induced apoptosis in the neonatal rat brain was examined by immunohistochemistry with the antibodies OX-6 and OX-42 which recognize MHC class II antigens and the CR3 complement receptor, respectively. Few OX-6-immunoreactive cells were observed in control rats, and in rats irradiated at postnatal day 2 and examined during the first 2 postnatal weeks. However, a transient increase in the number of OX-42-immunoreactive amoeboid microglia, containing large numbers of apoptotic cells, occurred at 6, 24 and 48 h after irradiation when compared with age-matched controls. These results show that X-ray-induced apoptosis promotes a short-lasting phagocytic response. PMID- 7478152 TI - Early amyloid-beta deposits show different immunoreactivity to the amino- and carboxy-terminal regions of beta-peptide in Alzheimer's disease and Down's syndrome brain. AB - That the topography, severity, and progression of beta-amyloid deposition in the brain of Alzheimer's disease (AD) and Down's syndrome (DS) cases is not uniform is well documented. We have addressed at present, the issue of whether the structural composition of beta-peptide (A beta) within the early amyloid deposits might contribute to this phenomenon. The cerebral cortex, the caudate/putamen, and the cerebellum from 10 AD and 8 DS cases were immunostained with antibodies that recognize the 1-17; 17-24 amino acid residues of A beta, and the COOH terminus of A beta 42 variant, thus to the epitopes of A beta located amino- and carboxy-terminally to the site of the putative alpha-secretase cleavage. We demonstrate that numerous diffuse, early plaques in AD and especially in DS cases show predominance of the carboxy-terminally located epitopes of A beta; the most prominent in the cerebellum, less pronounced in the cerebral cortex, and only marginal, or absent in the striatum, except for some DS cases. These data suggest that the deposition of the carboxy-terminal fragment of A beta truncated at the position of alpha-secretase cleavage or close to it in diffuse plaques may be brain-region-specific, reflecting either dissimilar processing of amyloid precursor protein or the resolution of early A beta deposits, and may substantially contribute to different progression of beta-amyloidosis in various brain regions. PMID- 7478154 TI - Distribution of neurons reactive for NADPH-diaphorase in the branchial nerves of a teleost fish, Gadus morhua. AB - The NADPH-diaphorase reaction was used to determine the distribution of postganglionic autonomic neurons in the branches of the glossopharyngeal and vagus nerves supplying the gill arches of the cod fish, Gadus morhua. Neurons were common in major nerve trunks in all gill arches, especially in the post trematic rami of the branchial nerves. From about 55% to more than 85% of the neurons in any branchial nerve were reactive for NADPH-diaphorase. The results suggest that the presence of NADPH-diaphorase, and presumably the ability to synthesise nitric oxide, have been a property of cranial parasympathetic neurons from early in the evolution of the vertebrates. PMID- 7478156 TI - Non-amoeboid locomotion of cultured microglia obtained from newborn rat brain. AB - The movements of cultured microglia obtained from newborn rat brain were examined by video enhanced-differential interference contrast (VEC-DIC) microscopy. Active microglia measured 9.03 +/- 1.06 microns in diameter (mean +/- SD, n = 33; range, 7.03-10.36 microns). The microglia appeared to become smaller with spread of lamellipodia. The short axis of actively moving microglia measured 7.03 +/- 0.49 microns (n = 7). The lamellipodia were thin, transparent and developed rapidly around the cell body (maximal speed of extension, 4 or 5 microns/s). When shear stress from the medium was applied to the surface of cultured cells, the microglia swam with flat lamellipodia serving as sails in the stream. Spontaneous non-amoeboid movements of microglia were observed: they pivoted, circled and marched in various directions using their lamellipodia. The angular speed of rotation was maximally 3 degrees/s. In 5 marching cells, the average speed (distance/s) was calculated at 1.01 +/- 0.54 microns/s (ca. 60 microns/min or 3.6 microns/h). PMID- 7478155 TI - Stimulation of glossopharyngeal and laryngeal nerve afferents induces expulsion only when it is applied during retching in paralyzed decerebrate dogs. AB - Stimulation of the oropharyngeal mucosa induces vomiting in humans, however, it has not been seen in animals. Thus, we attempted to induce fictive vomiting by stimulation of lingual and pharyngeal branch afferents of the glossopharyngeal nerve, and afferents of the superior and recurrent laryngeal nerves in paralyzed decerebrate dogs. The stimulation did not induce fictive vomiting when it was applied during eupnea, but induced fictive expulsion when it was applied during fictive retching. Threshold intensity for the induction decreased with the progress of retching. PMID- 7478157 TI - Sleep patterns in cyclic and pseudopregnant rats. AB - In order to clarify the relationship between sleep and reproductive activities, time-course changes in sleep were analyzed in normal female rats. The state of sleep-wakefulness was continuously monitored for 4 weeks including two consecutive 4-day estrous cycles, a 12-day pseudopregnant period and a subsequent 4-day cycle. Sleep patterns in estrous cycle were characterized by a marked reduction in nocturnal non-rapid-eye-movement sleep (NREMS) and rapid-eye movement sleep (REMS) at proestrus. A significant increase in nocturnal NREMS and REMS occurred immediately after the induction of pseudopregnancy by sterile mating, lasting for the whole pseudopregnant period for NREMS and during the early and mid period for REMS. In contrast, diurnal REMS tended to decrease towards the end of pseudopregnancy. The circadian rhythm of brain temperature exhibited no reproductive activity-dependent change. The dynamic changes in sleep may correlate alterations in neuroendocrine activities specific to estrous cycles and pseudopregnancy. PMID- 7478158 TI - Transforming growth factor-beta 1 levels are elevated in the striatum and in ventricular cerebrospinal fluid in Parkinson's disease. AB - Transforming growth factor (TGF)-beta 1 content was measured for the first time in the brain (caudate nucleus, putamen, and cerebral cortex) and in ventricular cerebrospinal fluid (VCSF) from control and parkinsonian patients by a sandwich enzyme immunoassay. The concentrations of TGF-beta 1 were significantly higher in the dopaminergic striatal regions in parkinsonian patients than those in controls, but were not significantly different in the cerebral cortex between parkinsonian and control patients. Furthermore, the concentrations of TGF-beta 1 in VCSF were significantly higher in parkinsonian patients than those in non parkinsonian control patients. Since TGF-beta 1 has potent regulatory activity on cell growth, these results suggest that TGF-beta 1 may have some significant modulatory role in the process of neurodegeneration in Parkinson's disease. PMID- 7478159 TI - Expression of Cu,Zn-superoxide dismutase mRNA after cold and contusion injury in the rat brain. AB - We analyzed the expression of Cu,Zn-superoxide dismutase (SOD) mRNA in both contusion and cold injury. Twenty-three rats were divided into 3 groups: a control group, a contusion group, and a cold injury group. Six hours after the injury, the rats were decapitated and the gray matter was resected from 3 portions: the core of the injured cortex, its periphery, and a distal portion on the non-lesion side. Based on the specific gravity of each sample, almost the same degree of edema developed in both injury groups. The mRNA expression in the cold injury group, however, significantly decreased in all portions. The extremely low temperature associated with cold injury is a possible cause of the decrease in Cu,Zn-SOD mRNA. PMID- 7478160 TI - Polymerase chain reaction quantification of lymphoid amyloid precursor protein mRNAs in Alzheimer's disease and Down's syndrome. AB - Amyloid beta-protein, the major constituent of Alzheimer's disease (AD) brain amyloid deposits, is encoded by several alternatively spliced amyloid precursor protein (APP) mRNAs. The well-established associated in Down's syndrome (DS) between APP overproduction and premature development of AD, as well as the recent demonstration of an increase in APP transcripts from lymphoblastoid cells of familial AD cases, suggest aberrant transcriptional regulation of some genes in AD. We assayed steady-state expression of the APP gene transcripts in peripheral blood mononuclear cells (PBMC) of AD and DS patients using quantitative polymerase chain reaction of reverse-transcribed mRNAs, and we compared their levels of PBMC APP expression with those of young and age-matched healthy controls. Results indicate APP mRNAs were of comparable abundance in PBMC obtained from 9 AD patients, 7 young controls and 12 age-matched controls. These data suggest regulation of APP mRNAs is normal in AD and DS PBMC. PMID- 7478161 TI - Cognitive potentials: ipsilateral corticocortical interconnections in prefrontal human cortex ablations. AB - The research deals with the possible role of the essentially monosynaptic bidirectional corticocortical connections between occipito-temporo-parietal association cortical areas and frontal areas in the genesis of some contingent negative variation (CNV) components, especially on the supramodal dorsolateral prefrontal regions. With standard and topographic mapping methods of analysis, the multicomponent CNV complex formation was examined in 7 patients with extensive frontal cortex ablations exactly identified through CT/MRI examinations, and in 10 normal subjects. On the scalp over the ablated frontocortical areas, no consistent post-warning auditory N100 a-b-c, P200, P300, early and late CNV components were recordable. The hypothesis is proposed that the bidirectional ipsilateral long-distance pathways which interconnect uni polymodal occipito-temporo-parietal cortical areas to prefrontal ones, in particular the arcuate-superior longitudinal and superior/inferior occipito frontal fasciculi, play an important role in the genesis of several CNV complex components, especially the multicomponent post-S1 auditory N100. The posteroanterior sequential latency differences of these neurocognitive components, roughly measured along the scalp or on MRI imagings, is probably accounted for by the transcortical ipsilateral conduction time of about 1 cm/ms (10 m/s). PMID- 7478162 TI - Morphological alterations in dendritic spines of rat hippocampal neurons exposed to N-methyl-D-aspartate. AB - Individual cultured rat hippocampal neurons grown on glass coverslips were loaded with lucifer yellow (LY) and segments of their dendrites, containing dendritic spines, were exposed to the glutamate agonist N-methyl-D-aspartate (NMDA). Repeated pulse application of NMDA caused a reduction of spine length by 20%, within 1-2 h of exposure to the drug. No such changes were seen in spines exposed to NMDA in presence of the antagonist 2-aminophosphonovalerate or in spines on sister dendrites which were not exposed to NMDA. It is suggested that dendritic spines shrink following exposure to excessive synaptic stimulation. PMID- 7478163 TI - VIP-like immunoreactive projections from the dorsal raphe and caudal linear raphe nuclei to the bed nucleus of the stria terminalis demonstrated by a double immunohistochemical method in the rat. AB - To localize the vasoactive intestinal polypeptide (VIP)-like immunoreactive neurons at the origin of the plexuses of VIP-immunoreactive fibers found in the bed nucleus of the stria terminalis (BNST), we combined iontophoretic injections of the retrograde tracer cholera toxin B-subunit (CTb) with VIP immunocytochemistry. Following CTb injections in the BNST, 2 groups of retrogradely labeled neurons immunoreactive to VIP were detected: (1) a dorso caudal group localized in the dorsal part of the dorsal raphe nucleus and the adjacent ventro-lateral part of the periaqueductal gray; and (2) a rostral group in the caudal linear raphe nucleus. No double-labeled cells were observed in other nuclei including those of the amygdala and the supramammillary nucleus. PMID- 7478164 TI - Attenuated inhibition by levofloxacin, l-isomer of ofloxacin, on GABA response in the dissociated rat hippocampal neurons. AB - The effects of ofloxacin (OFLX) and its isomers, levofloxacin (LVFX) and DR-3354, on the gamma-aminobutyric acid (GABA)-induced Cl- current in acutely dissociated rat hippocampal CA1 neurons were investigated using nystatin perforated patch recording configuration under voltage-clamp conditions. At 10(-5) M these 3 compounds themselves did not affect the GABA response. Biphenylacetic acid (BPA) at 10(-5) M also had no effect on the GABA response, but BPA greatly suppressed the GABA response in combination with these 3 compounds without affecting the reversal potential of GABA response. The inhibitory effects of OFLX and DR-3354 on the GABA response were stronger than that of LVFX. LVFX inhibited the response in a competitive and voltage-independent manner. The results suggest that LVFX has lower CNS adverse effects, such as convulsions, compared to OFLX. PMID- 7478165 TI - Arachidonic acid and lysophosphatidylcholine modulate excitatory transmitter amino acid release from the rat cerebral cortex. AB - The involvement of phospholipase A2 (PLA2) products in ischemia-evoked release of excitatory neurotransmitter amino acids (EAAs) from the cerebral cortex was studied in a four vessel occlusion rat model of cerebral ischemia/reperfusion. In comparison with untreated animals, arachidonic acid (AA; 5 x 10(-7) M to 5 x 10( 5) M) significantly reduced the ischemia-evoked efflux of glutamate and aspartate into cortical superfusates. Direct application of lysophosphatidylcholine (LysoPC; 55.4 micrograms/ml) to the cerebral cortex of non-ischemic animals resulted in a significant increase in glutamate levels. These results indicate that the immediate products of PLA2 action on plasma membrane phospholipids can either enhance or inhibit excitotoxic amino acid release following cerebral ischemia. The effect of AA is likely to be a result of its ability to inhibit PLA2; that of LysoPC, a consequence of its detergent action. PMID- 7478166 TI - Co-localization of angiotensin II and gamma-aminobutyric acid in axon terminals in the rat subfornical organ. AB - Angiotensin II (Ag II) and gamma-aminobutyric acid (GABA) in the subfornical organ have been implicated in drinking and cardiovascular responses to changes in circulating hormones. We combined immunogold silver labeling of Ag II with immunoperoxidase detection of GABA to determine whether there might be common cellular sites for their physiological actions in this circumventricular region. Electron microscopy showed that numerous terminals in central portions of the rat subfornical organ contained both Ag II and GABA immunoreactivity. These terminals as well as others exclusively labeled for AgII or GABA formed mainly symmetric, inhibitory type synapses with unlabeled dendrites. Immunogold-silver aggregates recognizing AgII were often detected near non-synaptic portions of the plasma membrane with or without apparent association with large dense core vesicles. In contrast, the GABA immunoperoxidase labeling was most intensely localized to membranes of small clear vesicles which were aggregated near the presynaptic junction. Our results indicate that in rat subfornical organ, neuronal AgII may modulate the inhibitory postsynaptic responses to GABA following release from single axon terminals. PMID- 7478167 TI - An adenosine triphosphate (ATP)-sensitive K+ channel of rat neocortical neurons is bi-gated by intracellular ATP and voltage: a novel channel gating mechanism? AB - To determine whether the adenosine triphosphate (ATP)-sensitive K+ (K-ATP) channels, which are suggested to be mainly regulated by intracellular ATP or other kinds of triphosphate nucleotides, are gated by membrane potentials (Vm), single K-ATP channel currents were studied on inside-out membrane patches of neurons acutely dissociated from Sprague-Dawley rat neocortex. The K-ATP channels recorded have a unitary conductance from 96.97 +/- 5.32 pS (n = 11) at potentials of approximately 10-60 mV to 98.31 +/- 3.26 pS (n = 11) at approximately -10 to 60 mV. Besides being inhibited by cytoplasmic ATP, channel kinetics was also affected by Vm. Open- and closed-time histograms were well fitted by 2 exponentials, suggesting that the channels have 2 open and closed states. Mean open time (tau om), open probability increased while mean closed time (tau cm) decreased with depolarization. The fitted equations of the relationships between Vm and those kinetic parameters may be described as: tau om = -159.26lnV + 403.64, Po = -0.01Vm2 + 0.08Vm + 0.87 and tau cm = 0.17Vm3-2.5Vm2 + 10.35Vm - 7.68, respectively. We suggest that the K-ATP channels be bi-gated by both intracellular ATP and membrane potentials. This property of the neuronal K-ATP channels may be related to their pathophysiological functions. PMID- 7478168 TI - Multiple protracted cauda equina constrictions cause deep derangement in the lumbosacral spinal cord circuitry in the dog. AB - Neuropathological changes of the neuronal pools and spinal cord circuitry in the lumbosacral segments were studied in a canine model of multiple protracted cauda equina constrictions. Anterograde degeneration of all sacrococcygeal and L7 dorsal root fibers was detected in S1-S3 and lower lumbar segments. A narrow degenerated gracile fascicle was found in all thoracic and cervical segments terminating in the gracile nucleus. Transneuronal degeneration of middle-sized and large neurons, located in S1-S3 and sporadically in L7 segments, was noted. Identical transneuronal degeneration was seen in a group of small neurons located in the ventralmost part of lamina VII in S1-S3 segments. Simultaneously, a terminal degeneration was detected in the lateral cervical nucleus and in the ventral posterior lateral nucleus of the ventrobasal thalamic complex. Concomitantly, a fully developed retrograde degeneration affecting motoneurons in the ventrolateral portion of the anterior horn in S1-S3 segments appeared. PMID- 7478169 TI - Ataxic breathing during alternating forearm movements of various frequencies in cerebellar patients. AB - Breathing was analyzed at rest and during sinusoidal tracking movements of the forearm of the dominant side at various frequencies in 11 patients with a cerebellar impairment and in 9 healthy subjects. In the patients, breathing movements were always rhythmical as in normal subjects during rest, but sometimes were interrupted by intermittent breathing-arrests during tracking (breathing ataxia). In 9 of the 11 cerebellar patients, the normal phase relationships between breathing and forearm movements at target frequencies in the spontaneous breathing rate range were absent. Thus patients with cerebellar lesions not only show disturbed skeletomotor movements, but also show an impaired coordination between breathing and limb movements. PMID- 7478170 TI - Electrophysiology and morphology of a new type of cell within layer II of the rat lateral entorhinal cortex in vitro. AB - Using a combination of intracellular recording and morphological techniques, we describe the properties of a new cell type within layer II of the lateral entorhinal cortex. A thick and bifurcating apical dendrite and thinner basal dendrites extended from the pyramidal shaped cell body. The axon ramified within all superficial layers of the lateral entorhinal cortex. These pyramidal-like cells exhibited 2 pronounced electrophysiological features; a high threshold for spike generation, and their prominent excitatory synaptic potentials with little inhibition following lateral entorhinal cortex stimulation. The electrophysiological properties and the axonal morphology suggest that this cell type has a local information processing role within the lateral entorhinal cortex. PMID- 7478171 TI - Theta power decreases in preparation for voluntary isometric contractions performed with maximal subjective effort. AB - In order to find EEG parameters that can be attributed to movements performed with maximal subjective effort, EEG recordings and force measurements were realized in connection with isometric contractions (IMC). IMC were performed with submaximal and maximal subjective effort. Mean spectral power density within the theta band was found as an indicator for maximal subjective effort. The theta power across the parieto-occipital area decreases from rest through movements performed with submaximal force to movements performed with maximal effort. It is argued that this theta decrease possibly reflects a down-regulation of the posterior attention system in order to minimize the influences of external stimuli during the preparation for voluntary IMC performed with maximal subjective effort. PMID- 7478172 TI - Identification of the alternative spliced form of the alpha 2/delta subunit of voltage sensitive Ca2+ channels expressed in PC12 cells. AB - The alpha 2/delta subunit of voltage sensitive Ca2+ channels expressed in PC12 has been cloned and partially sequenced. The message observed in Northern blot analysis displays a 7.5 kb transcript, identical in size to mRNA of rabbit skeletal muscle and rat brain. The nucleotide sequence of the cloned alpha 2 subunit of the PC12 specific cDNA is > 99% identical to rat brain sequence and 85% to skeletal muscle. Reverse-transcriptase-polymerase chain reaction (RT-PCR) of the alternative splicing region identifies two deleted regions of 57 bp and 21 bp in PC12 expressed alpha 2/delta transcript. The alternative variant alpha 2e of alpha 2/delta subunit which is expressed in PC12 cells was previously identified in human embryonic kidney (HEK293) cells. RT-PCR analysis show two different sized alternative PCR fragments in rat lung and none in rat spleen, kidney and intestine. Antibodies prepared against a 19 amino acid peptide within the alternative spliced region effectively inhibits [3H]dopamine release in PC12 cells. This implies that the alternatively spliced region is positioned extracellularly and is involved in regulation of the L-type Ca2+ channel-mediated transmitter release. PMID- 7478173 TI - Bradykinin depolarises the rat isolated superior cervical ganglion via B2 receptor activation. AB - Experiments were undertaken to characterise the action of kinins on sympathetic neurones of the rat superior cervical ganglion (SCG) by use of in vitro grease gap, extracellular recording techniques in conjunction with selective agonists and antagonists for B1 and B2 bradykinin (BK) receptors. Superfusion of BK (10 nM to 10 microM) to the ganglion produced a concentration-related depolarisation (pD2 = 7.02 +/- 0.04, n = 7) which was inhibited by the selective B2 antagonist HOE 140 (10-100 nM), but not by the B1 antagonist Leu8desArg9 BK (1 microM), indomethacin (7 microM) or the nitric oxide synthase inhibitor L-NG-nitroarginine methyl ester (300 microM). DesArg9BK (10 nM to 10 microM) had no effect on membrane potential. Pre-treatment of animals with intravenous bacterial lipopolysaccharide (LPS, 3 mg kg-1) failed to induce B1 receptor-mediated depolarisations of SCG neurones, or change responses to BK (P > 0.05, n = 4). These experiments highlight and characterise the action of BK as a neuromodulator of sympathetic neurones via B2 receptor activation. PMID- 7478175 TI - Reduced mitochondrial respiration in mouse cerebral cortex during chronic hypoxia. AB - Respiratory activity and NADH CoQ reductase (complex I) and cytochrome c oxidase (complex IV) activities were measured in free (non-synaptosomal) mitochondria isolated from cerebral cortex of male Balb/c mice exposed to intermittent hypobaric hypoxia (450 Torr; 4300 m) for 21 days and compared to normoxic (sea level) controls. In the hypoxic we found a 47% reduction of oxygen uptake during state 3 (ADP and substrate present), 12% reduction during state 4 (no ADP present) and 20% reduction in the uncoupled respiration rate with pyruvate plus malate as substrates. Respiratory control ratio (RCR) decreased by 24%. No change in the ADP/O ratio was seen. NADH CoQ reductase activity decreased by 30% and cytochrome c oxidase by 17%, suggesting that under conditions of chronic hypoxia, the reductions of mitochondrial respiratory activities are caused, at least in part, by enzymatic alterations of the electron transport chain (complex I and complex IV). The decreased activity of these enzymes could contribute to alterations in neuronal activity by reducing brain energy metabolism during development under conditions of chronic hypoxia. PMID- 7478174 TI - Inhibition by dihydropyridines of regulatory volume decrease and osmolyte fluxes in cultured astrocytes is unrelated to extracellular calcium. AB - The 1,4-dihydropyridines (DHP), nimodipine (NMD) and nitrendipine (NTD) were potent blockers of regulatory volume decrease (RVD) and the volume-associated release of [3H]taurine and chloride (measured as 125I) in 2-weeks cultured rat cerebellar astrocytes. The IC50 were 30 microM and 29 microM for taurine efflux and 26 and 27 microM for C1 efflux for NMD and NTD, respectively. Inhibition by DHP was independent of extracellular Ca, as the effect was the same in media with 1 mM Ca or without Ca and 0.5 mM EGTA. DHP did not affect the basal (isosmotic) release of [3H]taurine or 125I inhibition by DHP (measured only on [3H]taurine efflux) was the same in 3-4 weeks cultured cerebellar astrocytes, 2-4 weeks cultured cortical astrocytes and 2-weeks cultured cerebellar astrocytes treated with dibutyril cAMP. Diltiazem (50 microM) and verapamil (100 microM) failed to inhibit RVD or osmolyte efflux. PMID- 7478176 TI - Sodium depletion and Fos-immunoreactivity in lamina terminalis. AB - Rats were either depleted of sodium by treatment with a diuretic or were made hypovolemic by injection of polyethylene glycol (PEG) and their brains were subsequently examined for induction of Fos-like immunoreactivity (FLI). FLI was observed in the lamina terminalis (LT) after both treatments; FLI was observed in the magnocellular neurosecretory hypothalamic areas only after PEG. Implications of these data for brain substrates of sodium appetite are discussed. PMID- 7478177 TI - Zn2+ potentiates steady-state ATP activated currents in rat nodose ganglion neurons by increasing the burst duration of a 35 pS channel. AB - Zn2+ and ATP are normally present in serum at low levels and are released from some neurons during synaptic activity. In whole-cell recordings, 10 microM Zn2+ potentiated steady-state currents in rat nodose ganglion nerve cells by 18% in 0.5 microM ATP and 120% in 2 microM ATP. Fluctuation analysis of whole-cell suggested the mechanism was increased burst duration. In single-channel recordings, 10 microM Zn2+ did not increase the ATP-activated channel conductance but did increase the mean burst duration of the 35 pS primary open conductance state. These observations indicate that Zn2+ potentiates the steady-state response to extracellular ATP by increasing the burst duration of the channels. PMID- 7478179 TI - Presentation rate and magnitude of stimulus deviance effects on human pre attentive change detection. AB - The present study investigated the effects of the inter-stimulus interval (ISI) and the magnitude of deviance on the mismatch negativity (MMN), an event-related potential index of pre-attentive change detection. The latency of the MMN to infrequent changes in intensity and duration was prolonged by increasing the ISI from 400 ms to 4 s. Results indicate that these features are preserved in sensory memory for several seconds. By increasing the ISI the sharpness of the neural traces decreases, thus prolonging the change detection process. PMID- 7478178 TI - alpha-Amino-3-hydroxy-5-methyl-4-isoxazolepropionate receptors modulate dopamine release in rat hippocampus and striatum. AB - The effects of infusing the glutamate receptor agonist alpha-amino-3-hydroxy-5 methyl-4-isoxazolepropionate (AMPA) on the release of dopamine (DA) has been studied in rat hippocampus and striatum in vivo. In hippocampus, AMPA (1-10 microM) produced a dose related increase in dialysate DA, but at 100 microM AMPA a sustained decrease in extracellular DA was observed. However, when samples were collected at 5-min intervals 100 microM AMPA infusion revealed a brief increase in hippocampal dialysate DA. Infusion of 100 microM AMPA and 500 microM diazoxide, which blocks AMPA receptor desensitization, led to a marked increase in extracellular DA, as did diazoxide alone, although to a lesser extent. The AMPA receptor antagonist 6-cyano-7-nitroquinoxaline-2,3,-dione (CNQX; 200 microM) reversed the effect of AMPA and/or diazoxide infusion on dialysate DA and when infused alone, CNQX also decreased hippocampal dialysate DA. AMPA (50-500 microM) increased striatal DA release. The effect of AMPA on extracellular DA was reversed by CNQX (200 microM). Diazoxide infusion caused a decrease in striatal DA release, and this was not affected by CNQX. These data suggest that hippocampal, but not striatal AMPA receptor desensitization may play a role in regulating DA release. PMID- 7478180 TI - Innervation-independent changes in the mRNAs encoding tyrosine hydroxylase and the norepinephrine transporter in rat adrenal medulla after high-dose reserpine. AB - To determine whether a trans-synaptic mechanism triggered the effects of reserpine on adrenomedullary mRNAs encoding the norepinephrine transporter and tyrosine hydroxylase, we administered 10 mg/kg reserpine to rats after unilateral splanchnicotomy, and examined their adrenal medullas using quantitative in situ hybridization. Splanchnicotomy did not alter the decrease in norepinephrine transporter mRNA that follows reserpine administration, but diminished the reserpine-induced increase in tyrosine hydroxylase mRNA by almost 80%. Despite the latter effect, reserpine still induced a significant increase in tyrosine hydroxylase mRNA in denervated adrenal medullas, compared to vehicle-treated adrenal medullas. These results show that a trans-synaptic mechanism does not trigger the decrease in adrenomedullary norepinephrine transporter mRNA following reserpine. In addition, an innervation-independent mechanism mediates a portion of the reserpine-induced increase in adrenomedullary tyrosine hydroxylase mRNA. PMID- 7478181 TI - Cerebrovascular parasympathetic innervation contributes to coupling of neuronal activation and blood flow in rat somatosensory cortex. AB - We measured the increase of regional cerebral blood flow (rCBF) in the somatosensory cerebral cortex occurring in response to a standard stimulation of the L. side mystacial vibrissae (facial whiskers) in rats anaesthetised with halothane, in conjunction with blocking of activity in the R. side parasympathetic (PS) and sensory fibres innervating the cerebral vessels. Blocking was achieved reversibly and repeatedly by means of a cooling probe. When the PS fibres and the nasociliary nerve (NCN) were blocked together, but not when the NCN was blocked alone, the R. side rCBF increase occurring with whisker stimulation was significantly reduced. Our results indicate that, in addition to the intrinsic cortical factors demonstrated in earlier studies, the cerebrovascular PS innervation, but not the NCN, contributes to the increase in cortical rCBF associated with somatosensory cortical neuronal activation. PMID- 7478182 TI - Non-involvement of the redox site of NMDA receptors in bidirectional synaptic plasticity in the CA1 area of the rat hippocampus in vitro. AB - We have examined the effects of the redox reagent 5,5'-dithiobis-2-nitrobenzoic acid (DTNB) on synaptic potentials recorded extracellularly from the CA1 area in hippocampal slices following low frequency stimulation (LFS) and tetanic stimulation (TS). Application of DTNB (200 microM) neither changed synaptic responses, nor prevented the expression of TS-induced long-term potentiation of synaptic responses and their depotentiation by LFS. Conversely, in naive slices, LFS still induced long-term depression of synaptic responses following application of DTNB. This depression could be subsequently reversed with a TS. It is concluded that the redox state of N-methyl-D-aspartate receptors does not affect the expression of long-term potentiation and depression of synaptic responses. PMID- 7478183 TI - Nitric oxide produced by activated astrocytes rapidly and reversibly inhibits cellular respiration. AB - Cultured astrocytes, activated to express the inducible form of nitric oxide synthase, produced up to 1 microM nitric oxide (NO) measured by a NO-selective electrode, while non-activated cells produced no detectable NO. The production of NO was associated with an inhibition of cellular respiration, measured simultaneously by an oxygen electrode. The inhibition of respiration was rapidly reversed by inhibiting the NO synthase or by binding the NO with haemoglobin. The respiratory inhibition had an NO, oxygen and substrate dependence consistent with NO-inhibition at cytochrome oxidase. This is the first demonstration that cells can reversibly inhibit mitochondrial respiration via NO production. This inhibition is large and potentially important in a range of pathophysiological conditions. PMID- 7478184 TI - Sensory denervation with capsaicin attenuates inflammation and nociception in arthritic rats. AB - The relatively few studies that have investigated the effects of the nervous system on chronic joint disease have reported conflicting results. We have reassessed the effects of capsaicin on experimental polyarthritis with particular reference to the relationship between changes in nociception and changes in process and outcome measures of disease activity. Capsaicin pretreatment significantly attenuated both joint swelling and disease outcome as determined by quantitative radiology and histology. There was a close correlation between process measures of inflammation and mechanical hyperalgesia in both the untreated and capsaicin treated arthritic groups. The results confirm a suppression of inflammation by capsaicin and imply that the nociceptive and pro inflammatory (neurogenic inflammation) activities of capsaicin-sensitive fibres are closely linked such that stimuli which cause pain will also induce neurogenic inflammation and vice versa. PMID- 7478185 TI - Cutaneous localisation of laser induced pain in humans. AB - Localisation of painful laser-induced stimuli has been compared to localisation of tactile stimuli in normal humans. Laser stimulation evoked pain of two qualities (sharp pricking pain and hot burning pain). Sharp pricking pain was found to be localised with almost equal precision to tactile stimuli (13.8 +/- 3.4 versus 11.6 +/- 2.3 mm) on the dorsum of the hand, (21.5 +/- 7.9 versus 20.6 +/- 7.5 mm) on the forearm and (15.5 +/- 5.6 versus 13.8 +/- 5.4 mm) on the foot, respectively. There was no significant difference between the ability to localise tactile stimuli and hot burning pain except on the dorsum of the hand. These results indicate that tactile information is not essential for the accurate localisation of cutaneous pain. PMID- 7478186 TI - SMI-32 antibody against non-phosphorylated neurofilaments identifies a subpopulation of cultured cortical neurons hypersensitive to kainate toxicity. AB - SMI-32, an antibody against a non-phosphorylated neurofilament epitope identifies a subpopulation of human cortical neurons preferentially lost in Alzheimer's or Huntington's disease. In murine cortical cultures SMI-32 labeled a small subset of neurons exhibiting enhanced vulnerability to kainate toxicity. Most SMI-32(+) neurons were GABAergic and exhibited kainate-activated Co2+ uptake. Thus expression of Ca2+ permeable AMPA or kainate receptor-gated channels likely underlies the heightened vulnerability of SMI-32(+) cortical neurons to kainate. PMID- 7478187 TI - Removal of hydroxyl radical increases nitric oxide generators-induced [3H]GABA release from mouse cerebral cortical neurons. AB - We investigated the effect of removal of hydroxyl radical on nitric oxide (NO) induced [3H]GABA release from cerebral cortical neurons. NO generators, S-nitroso N-acetylpenicillamine and sodium nitroprusside, significantly increased [3H]GABA release, which was completely abolished by hemoglobin. These results indicate that the release of [3H]GABA evoked by these NO generators is mediated by NO formation. N,N'-Dimethylthiourea (DMTU), a hydroxyl radical scavenger, dose dependently stimulated the increase of NO-induced [3H]GABA release. Similarly, other hydroxyl radical scavengers such as uric acid and mannitol, significantly enhanced the NO-induced [3H]GABA release. On the other hand, each hydroxyl radical scavenger alone showed no effect on [3H]GABA release in the absence of NO generators. These results indicate that the removal of hydroxyl radical enhances NO-evoked [3H]GABA release from cerebral cortical neurons. PMID- 7478188 TI - Bimodal patterns of human melatonin secretion consistent with a two-oscillator model of regulation. AB - In many animals, changes in duration of nocturnal melatonin secretion chemically mediate effects of seasonal changes in nightlength on behavior and physiology. According to one model, the changes in duration of secretion result from adjustments in the timing of two circadian oscillators, one entrained to dusk, controlling onset, and another entrained to dawn, controlling offset. Consistent with this model, in six women, we found separate and reproducible evening and morning peaks of melatonin secretion that might represent the separate expressions of rhythms of two oscillators. PMID- 7478189 TI - Induction of nitric oxide synthase and motoneuron death in newborn and early postnatal rats following spinal root avulsion. AB - It is well established that immature motoneurons are more vulnerable to axonal injury than the adult ones. Many previous studies used distal axonal injury, such as sciatic nerve transection, to examine the response of immature motoneurons to injury. In the present study, a more severe injury, spinal root avulsion, was performed in newborn and early postnatal rats, and the response of immature spinal motoneurons to the injury was observed. In newborn rats, root avulsion causes motoneuron loss which is similar to that of previous studies by distal axonal injury. During the early postnatal development, motoneuron loss in immature rats is greater following root avulsion when compared to cell loss following distal axonal injury at any given age. Following root avulsion, nitric oxide synthase (NOS) is induced in injured immature motoneurons. The induction and accumulation of NOS in injured motoneurons is much more rapid in the immature animals than in the adult, which is coincident with the more rapid motoneuron loss in the immature animals. Results of the present study indicate an involvement of NOS in neuronal degeneration. However, the precise role of NOS in the response of motoneurons to axonal injury is not clear and needs to be further studied. PMID- 7478190 TI - 2-[125I]iodomelatonin binding sites in the bovine hippocampus are not sensitive to guanine nucleotides. AB - The discrete distribution and pharmacological characteristics of melatonin binding sites in the bovine hippocampus were determined. Autoradiography revealed the presence of melatonin binding sites in the stratum lacunosum-molecularis of the hippocampus (CA1), stratum molecularis of the subiculum and in the enthorhinal cortex. Analysis of the kinetic parameters demonstrated that the binding was stable and reversible, represented by a single class high affinity binding sites (Kd 40 pM, Bmax = 3.9 fmol/mg protein). However, 2-iodomelatonin and 2-bromomelatonin inhibited 2-[125I]iodomelatonin binding in a biphasic manner. The presence of 4 mM CaCl2 did not cause changes in the affinity constant values. Finally, experiments performed with GTP gamma S revealed that binding affinity was not decreased even with high concentrations of the nucleotide. These findings show that 2-[125I]iodomelatonin binding sites in the bovine parahippocampal-hippocampal region possess some binding features not common to melatonin receptors described so far; moreover they seem not to be linked to a regulatory G-protein. PMID- 7478191 TI - Loss of p75 nerve growth factor receptor mRNA containing neurons in rat forebrain after intraventricular IgG 192-saporin administration. AB - Cholinergic neurons of the basal forebrain express the p75 (low affinity) nerve growth factor receptor (NGFr) on the cell surface. It has been previously shown that an immunotoxin that recognizes the p75 NGFr, IgG 192-saporin, eliminates these neurons as judged by a variety of techniques after intraventricular injection into rat brain. Here we show that this loss of neurons can also be identified by detecting the disappearance of the p75 NGFr mRNA utilizing a non radioactive in situ hybridization method. PMID- 7478192 TI - Decline and preservation of reversal learning abilities and acquisition in the course of senescence. AB - Different types of learning and memory functions decrease at different rates in senescence. The present study examines which types of mental functions show a relatively early decline and which learning abilities are relatively preserved in late senescence by investigating different types of learning abilities in water maze tests. Two groups of senescent male Wistar rats aged 24 months (group W24) and 30 months (group W30), respectively, were compared to adult rats (12 months, group W12). Group W24 represents 'senescent' and group W30 'late-senescent' rats. Whereas acquisition showed a relatively late decline (in group W30), reversal learning was impaired relatively early (group W24). PMID- 7478193 TI - Increase in plasma nitric oxide end products following rat cortical injury. AB - The changes in plasma nitric oxide (NO) end products, nitrite (NO2-) and nitrate (NO3-), were studied following cortical injury in rats. At 3 days after stereotactic cortical injury (day 3), plasma NO end products were significantly increased (P < 0.01), and decreased by day 7. This increase on day 3 was inhibited by a selective inhibitor of inducible NO synthase (NOS), aminoguanidine (100 mg/kg, i.p. on days 1 and 2, P < 0.001). The present study first demonstrated the temporary increase in plasma NO end products, which is attributable to the inducible NOS activation after cerebral injury. PMID- 7478194 TI - Long-term potentiation at low temperature is stronger in hippocampal slices from hibernating Turkish hamsters compared to warm-acclimated hamsters and rats. AB - Tetanus-induced long-term potentiation (LTP) of population action potentials at 22 degrees C was investigated in area CA1 of hippocampal slices prepared from hibernating (HH) and warm-acclimated Turkish hamsters (WH) and rats. LTP elicited at this temperature was significantly stronger in HH slices compared to WH and rat slices. Hibernation-related improvement of the ability to develop long lasting enhancement of synaptic transmission at low temperatures is interpreted as supporting hippocampal function during arousal from hibernation. PMID- 7478195 TI - B-50 (GAP-43) in the rat spinal cord caudal to hemisection: lack of intraspinal sprouting by dorsal root axons. AB - The controversial hypothesis that intraspinal sprouting by dorsal root axons promotes reinnervation of partially denervated neurons caudal to a low thoracic cord hemisection was re-investigated in rats using quantitative immunohistochemical analysis of the neural specific growth-associated protein B 50 (GAP-43) at postoperative survival times of 3, 10, 21, 42, and 90 days. The lack of increase in B-50-immunoreactivity in all segments below the hemisection at all survival times does not support the concept of intraspinal sprouting following the removal of supraspinal descending pathways. PMID- 7478196 TI - Betel chewing increases the skin temperature: effects of atropine and propranolol. AB - Although betel chewing produces sweating, facial flush and a warm sensation of the body, objective documentation of the temperature effect has not been available. In this report, the skin temperature of the ear and forehead was recorded before and during betel chewing in healthy betel chewers. Betel chewing produced a mean temperature increase of 2 degrees C and 0.5 degree C, respectively, for the ear and forehead. This hyperthermic response was almost completely abolished by atropine and partially inhibited by propranolol. The present data suggest that both sympathetic and parasympathetic mechanisms are involved in the skin thermal response to betel chewing. PMID- 7478197 TI - Numerous glial fibrillary tangles in oligodendroglia in cases of subacute sclerosing panencephalitis with neurofibrillary tangles. AB - Both neurons and oligodendroglia are preferentially infected in subacute sclerosing panencephalitis (SSPE). Massive argyrophilic and tau-positive glial fibrillary tangles (GFT) were found in oligodendroglia in two autopsy cases of SSPE with neurofibrillary tangles (NFT). GFT shared common phosphorylated tau epitopes with NFT, but were negative for ubiquitin. Electron microscopically, GFT consisted of compact bundles of irregularly woven tubules. Thus, GFT in SSPE differed from NFT showing regular constriction of tubules and from GFT in some other cytoskeletal disorders in which GFT reportedly consisted of straight tubules. PMID- 7478198 TI - Molecular analysis of three cases with hereditary motor and sensory neuropathy with myelin outfolding. AB - We describe three patients affected by a congenital motor and sensory neuropathy with excessive myelin outfoldings (MOs) [15]. Clinical and electrophysiological features supported the diagnosis of hereditary motor and sensory neuropathy. We previously reported a genetic study on these three patients, which failed to demonstrate either the duplication in chromosome 17p11.2 or the mutations at exons 1 and 2 of the peripheral myelin protein gene (PMP-22) and suggested an autosomal recessive (AR) inheritance. In this study we described the absence of the most common mutations, which characterized other forms of hereditary motor and sensory neuropathy (HMSN). In particular the absence of molecular changes in the PMP-22 gene definitively sets HMSN with MOs apart from the more common CMT1A, hereditary neuropathy with liability to pressure palsies (HNPP) and progressive sensory-motor polyneuropathy with tomaculous changes at sural nerve biopsy. PMID- 7478199 TI - A new animal model of cerebral infarction: magnetic embolization with carbonyl iron particles. AB - A new animal model of cerebral infarction was developed by magnetic embolization with carbonyl iron particles. An electromagnet was placed upon the lateral portion outside the orbit of Mongolian gerbils and charged with a current intensity of 2 A, 3 V (400 G) for 10 min. By intracardiac injection of iron particles at the beginning of the electric charge, infarction was selectively produced in the cerebral cortex. Two days after the operation, microhemorrhaging was seen in the necrotic region. Multiple scattered infarctions accompanied with microhemorrhagic signs closely resembled embolic infarction encountered in the human brain. PMID- 7478200 TI - Enhanced expression of tyrosine hydroxylase and aromatic L-amino acid decarboxylase in cerebellar Purkinje cells of mouse after hyperosmotic stimuli. AB - We examined by immunohistochemistry the effect of salt loading on the expression of tyrosine hydroxylase (TH), aromatic L-amino acid decarboxylase (AADC), and GTP cyclohydrolase I (GCH) in Purkinje cells of the mouse cerebellum. In control mice, only a few Purkinje cells were positive for TH or AADC. No Purkinje cells were stained for GCH. Drinking 2% sodium chloride for 2 weeks resulted in an increase in the number of TH- or AADC-positive Purkinje cells in the caudal vermis, paraflocculus and flocculus of the cerebellum. In contrast, no Purkinje cells were immunoreactive to GCH or L-DOPA after the salt loading. The present findings suggest that the salt loading differentially affects the expression of TH, AADC and GCH in Purkinje cells of the mouse cerebellum. PMID- 7478201 TI - Apolipoprotein E and B alleles in Parkinson's patients. AB - Parkinson's patients were genotyped for the apolipoprotein E alleles as well as polymorphisms at the apolipoprotein B loci to determine whether they were at risk for late onset Alzheimer's disease or coronary disease. The Parkinson's patients were at no greater risk for either disease than were the control spouses. The frequency for the APOE4 allele was 11% compared with the spouses, 10%. Interestingly, 20% of the patients had the 2/3 genotype which may have a protective effect from late onset Alzheimer's disease. PMID- 7478202 TI - Expression of interleukin-6 (IL-6) and IL-6 receptor mRNAs in rat adrenal medulla. AB - Using in situ hybridization and reverse transcription combined with the polymerase chain reaction (RT-PCR) we have studied the expression and developmental regulation of interleukin-6 (IL-6) and its receptor mRNAs in rat adrenal medulla. Labeling for both transcripts was much more intense over the adrenal medulla than over cortical regions at postnatal day 70. Levels of mRNAs of both genes measured by RT-PCR increased from birth to adulthood. In vitro survival and neurite growth of chromaffin cells from early postnatal rats were not affected by IL-6. Even so, the presence and developmental regulation of IL-6 and its receptor are consistent with a possible auto- or paracrine role of IL-6 in rat adrenal medulla. PMID- 7478203 TI - Specific binding of [3H]imipramine indicates the presence of a specific serotonin transport system on endothelial cells of porcine brain. AB - The blood-brain barrier (BBB) represents a complex epithelial interface in vertebrates that separates the blood compartment from the extracellular fluid compartment of the brain. Isolated microvessels are a tool to study the function of this interface in vitro. Here we report on attempts to demonstrate the presence of the serotonin transporter on microvessels from the porcine brain. For comparison, membrane preparations of brain tissue were used. The enrichment of the microvessel fraction determined by measurement of alkaline phosphatase activity was about 30-fold. In saturation experiments high- and low-affinity binding of [3H]imipramine could be demonstrated on brain microvessels. Different concentrations of unlabelled imipramine were used to inhibit the binding of [3H]imipramine in brain tissue and microvessels. Comparison of both preparations revealed a two-fold higher density of the high-affinity binding site, while the density of the low-affinity binding site was 28-fold higher in brain microvessels. Imipramine binding could be inhibited by potent non-tricyclic inhibitors of the serotonin transporter such as paroxetine and fluoxetine but also by the tricyclic antidepressant drugs clomipramine and desipramine. Therefore, it is concluded that [3H]imipramine labels serotonin uptake sites localized on porcine brain microvessels. PMID- 7478204 TI - Human serum albumin enhances sodium currents in NH15-CA2 neuroblastoma x glioma hybrid cells. AB - The effect of human albumin on neuronal voltage-dependent Na+ channels was studied in NH15-CA2 neuroblastoma x glioma hybrid cells. At an albumin concentration of 32 g/l, the transient Na+ currents, elicited at 1 Hz and recorded with the whole-cell technique, were increased to 125% the control value. The effect was complete within 2-4 s of extracellular albumin application; 35 s after washout of the protein it was not yet fully abolished. The albumin effect is mediated by a shift of the voltage dependence of steady-state inactivation of the Na+ channels towards more positive potentials. The activation of the channels is not noticeably influenced. PMID- 7478205 TI - Effect of local cholecystokinin-8 administration on extracellular levels of amino acids and glycolytic products monitored by in vivo microdialysis in the fronto parietal cortex of the rat. AB - The effects of local cholecystokinin-8 (CCK-8) administration on cortical extracellular levels of amino acids, catecholamines and metabolism products were studied in the halothane anaesthetized rat by in vivo microdialysis. CCK-8 (10 microM), administrated via a microdialysis probe, produced a significant increase in the levels of aspartate, glutamate and gamma-aminobutyric acid (GABA), but not of 3,4-dihydroxyphenylacetic acid (DOPAC), homovanillic acid (HVA), lactate and pyruvate, which were simultaneously monitored. The increase in aspartate and glutamate levels produced by CCK-8 was about 3-4-fold. The effect of CCK-8 on aspartate levels was significantly inhibited by the CCKB antagonist, L-365,260 (20 mg kg-1, s.c.), but not by the CCKA antagonist, L-364,718 (20 mg kg-1, s.c.). In contrast, the increase in glutamate levels was inhibited by both L-365,260 and L-364,718. GABA was slightly, but significantly increased (approximately 30%), by local CCK-8 and was inhibited by both CCK antagonists. The present results show that CCK-8 exerts a strong modulatory action on both aspartate and glutamate release in rat cortex. While the effect of CCK-8 on aspartate is selectively mediated via CCKB receptor subtype, the effect of CCK-8 on glutamate appears to be mediated via both CCKA and CCKB receptor subtypes. PMID- 7478206 TI - Tyrosine hydroxylase-containing fibres extend from the rat corpus striatum into grafts of muscularis externa and myenteric plexus. AB - Intrastriatal grafts of myenteric plexus produce a vigorous sprouting response in the surrounding rat brain. Since the striatum receives profuse dopaminergic innervation from the substantia nigra, we have investigated whether central catecholaminergic neurons participated in the sprouting response and grew into grafts of adult myenteric plexus (surrounded by smooth muscle) implanted in the adult corpus striatum. Three weeks after implantation, tyrosine hydroxylase containing fibres were observed to have grown into, and ramified within, the grafts. The extent of innervation was increased 6 weeks after implantation, and was not diminished if the superior cervical ganglia were removed (to destroy any fibres of sympathetic origin). PMID- 7478207 TI - Ubiquitinated inclusions in inclusion-body myositis patients are immunoreactive for cathepsin D but not beta-amyloid. AB - The nature of the inclusions in the human muscle disease inclusion-body myositis (IBM) has been the subject of debate. Parallels with Alzheimer's disease have been drawn after these inclusions were found to be ubiquitinated, and immunoreactive with antibodies to beta-amyloid (A beta) and certain amyloid associated proteins. We have used a battery of antibodies against A beta and associated proteins to immunostain muscle biopsies from patients with IBM. Although the inclusions are ubiquitinated, we could not show immunoreactivity for A beta or the associated proteins investigated. We did, however, find that the ubiquitinated inclusions colocalised with the lysosomal marker, cathepsin D. PMID- 7478208 TI - Morphine produces a biphasic modulation of substance P release from cultured dorsal root ganglion neurons. AB - We have previously reported that morphine produces a concentration-dependent multiphasic modulation (inhibitions and facilitations) of substance P (SP) release from trigeminal nucleus caudalis slices by activation of distinct populations of mu-, delta- and kappa-opioid receptors. In the present study, we have examined a wide range of morphine concentrations on K(+)-evoked SP release from dissociated rat dorsal root ganglion (DRG) neurons in culture. SP immunoreactivity was measured in the release buffer. Morphine produced a biphasic effect on K(+)-evoked SP release without affecting basal release. A concentration of 30 nM morphine facilitated SP release while a concentration of 1 microM suppressed release. Higher concentrations of morphine (10-30 microM) did not alter SP release. The facilitatory effect evoked by 30 nM morphine was abolished by opioid-receptor blockade with naloxone (30 nM) and the inhibitory effect produced by 1 microM morphine tended to be reversed. We conclude that an intact neuronal circuitry is not required for morphine to produce an opioid receptor mediated biphasic modulation of SP released from unmyelinated primary afferents. It is plausible that the dose-dependent biphasic effects of opioid agonists may also produce biphasic effects on nociception. PMID- 7478210 TI - Behavioural deficits in adult rats following long-term adrenalectomy. AB - Long-term adrenalectomy results in loss of neurones in the hippocampal formation of the adult rat. The effects of long-term adrenalectomy on spatial learning and exploratory behaviour in adrenalectomized (ADX), adrenalectomized normal weight gain (ADXNW), sham operated and naive control male Sprague-Dawley rats were investigated in this study. The ADX rats had significantly longer latencies in the Morris maze task compared to the other groups. In the open-field situation, as a novel finding, the ADX rats showed significantly lower rearing scores compared to other groups. These data indicate that long-term adrenalectomy causes impairment in spatial learning and explorative behaviour in the rat. PMID- 7478209 TI - Continuous infusion of acidified saline around the rat sciatic nerve produces thermal hyperalgesia. AB - Recent observations using both clinical and animal models have suggested that acidosis may initiate pain and sensitization. In the present study, we examined if changing the acidic environment around the rat sciatic nerve resulted in thermal hyperalgesia. Fresh solutions of preservative-free saline (PFS) and unbuffered PFS acidified to pH 3.5 were continuously infused around the left rat sciatic nerve for 7 days. Rats receiving unbuffered, acidified PFS developed a progressive thermal hyperalgesia that was maximal on infusion day 6. Unbuffered, acidified PFS significantly decreased the perineural pH (pH 6.9 +/- 0.15, P < 0.05), and decreasing perineural pH values were significantly correlated with increasing thermal hyperalgesia (r = 0.91) for individual rats. While it is likely that multiple factors play a role in the development of neuropathic pain, these data demonstrate that an acidic environment around the sciatic nerve will produce thermal hyperalgesia. PMID- 7478211 TI - Nitric oxide inhibition of the depolarization-evoked glutamate release from synaptosomes of rat cerebellum. AB - Effects of nitric oxide (NO) on release of amino acid transmitter were investigated by superfusion of synaptosomes prepared from rat cerebellum. After constant basal levels of amino acid release were obtained, exposure to a depolarizing concentration of KCl (30 mM) evoked 4.05-, 2.18- and 3.00-fold increases in release of glutamic acid (Glu), aspartic acid (Asp) and gamma aminobutyric acid (GABA) from synaptosomes. The perfusion with NO-donors inhibited the evoked increases in release of Glu and Asp in a concentration dependent manner, but not that in GABA release. A membrane-permeable analog of cyclic GMP, but not that of cyclic AMP, caused a similar reduction in the evoked release. The concentration of nitroprusside to increase cyclic GMP levels corresponded to that of nitroprusside to reduce the evoked release. These data suggest that NO may directly act upon the nerve terminals to inhibit release of excitatory amino acid transmitters. PMID- 7478212 TI - Endothelin-1 stimulates myristoylated alanine-rich C-kinase substrate (MARCKS) phosphorylation in rat cerebellar slices. AB - Protein phosphorylation induced by endothelins has been studied using [32P]orthophosphate-prelabelled rat cerebellar slices. Endothelin-1 increased phosphorylation of an 87 kDa protein in a time-dependent manner (reaching a maximum effect at about 2.5 min) and with an EC50 equal to 93 +/- 32 nM. Endothelin-3 and sarafotoxin 6c induced similar levels of phosphorylation. Endothelin-1 also promoted [3H]inositol phosphate accumulation with similar EC50 (71 +/- 7.5 nM). The phosphoprotein of 87 kDa seems to be myristoylated alanine rich C-kinase substrate (MARCKS) as demonstrated by acetic acid extraction. In addition, 12-O-tetradecanoylphorbol-13-acetate (TPA) increased 87 kDa protein phosphorylation while Ro-31-8220, a specific protein kinase inhibitor, inhibited both TPA and endothelin-induced 87 kDa protein phosphorylation. Therefore, it is concluded that protein kinase C is involved in the endothelin action on cerebellum. PMID- 7478213 TI - Protection against noise trauma by sound conditioning in the guinea pig appears not to be mediated by the middle ear muscles. AB - The aim of this study was to test whether the middle ear muscles (MEM) play a significant role in the phenomenon of sound conditioning in guinea pigs. After inducing a temporary threshold shift by noise exposure (2767 Hz tone, 103 dB SPL, 5 min), the magnitude and duration of loss, as well as the rate of recovery of the amplitude of the distortion product emission was determined at 1.75, 2.2, 2.8, and 3.5 kHz followed for 90 min post-exposure for (1) a sound conditioned group with intact MEM, and (2) a sound conditioned group with paralyzed MEM. Significant differences were not found for any of the distortion product parameters tested. The results suggest that the MEM do not significantly contribute to protection against noise trauma by sound conditioning. PMID- 7478214 TI - Muscle activity is different for humans performing static tasks which require force control and position control. AB - Muscle activation levels in humans were examined during two different static tasks which required the same joint angles and the same joint moments. In the isometric case, joint angles were fixed and subjects were required to match forces. In the isoinertial case, a constant load was imposed across the joint and the subject was required to match position. It was observed that for a specified posture and for specified load conditions, EMG activity varied depending on whether the limb was loaded isometrically or isoinertially. That is, different co activation relationships were observed for position control versus force control tasks during otherwise similar conditions. These results imply that the neural command for static tasks depends on more than joint angles and load magnitude. PMID- 7478215 TI - Long-term bFGF neuronal culture: reintroduction into serum medium yields neurons and non-neuronal cells with neuronal characteristics. AB - The potential use of bFGF immortalized cells as hosts for delivering foreign genes into nervous tissue led us to examine the effect of maintaining, E-18 hippocampal neurons for extended periods in bFGF culture prior to transfer into a standard, serum-containing, medium. We found: (1) many, if not most, precursors seen in bFGF, mature into glia and not into primary neurons after medium exchange; (2) the electrophysiology of the neurons which do mature after medium transfer and replating, is similar to that of neurons in standard cultures; (3) extended culture in bFGF prior to cell harvesting and replating into standard medium generates neurons from the precursors that possess proper neuronal polarization, morphology, and electrophysiology; and (4) extended bFGF also induces the expression, on transfer into standard medium, of an additional cell type with a distinct non-neuronal morphology that stains with the neuronal marker MAP-2. These results illustrate the need for additional characterization of long term growth factor effects on maintained progenitor cells prior to their use in gene therapy and transplantation. PMID- 7478216 TI - Vestibular experiences of humans during brief periods of partial sensory deprivation are enhanced when daily geomagnetic activity exceeds 15-20 nT. AB - The intensity of reported vestibular experiences by normal volunteers (n = 127, over a 4-year period) increased significantly in a step-like manner during partial sensory deprivation when the daily geomagnetic activity exceeded about 15 nT; the effect size was equivalent to a correlation of about 0.33. Post hoc analyses indicated that the geomagnetic activity during the 3-h period at the beginning of the previous sleep cycle was the single greatest contributor. The enhanced occurrences of experiences that are similar to those associated with complex partial epileptic seizures suggests that specific stimuli associated with geomagnetic activity above about 15 nT affects specific regions of the human brain; the vulnerability occurs during the subsequent 24 h. However, in non epileptic individuals these experiences must be amplified neurocognitively by removing input from auditory and visual modalities. PMID- 7478217 TI - Elevated striatal Fos immunoreactivity following 6-hydroxydopamine lesioning of the rat is mediated by excitatory amino acid transmission. AB - Pharmacological depletion of dopaminergic neurotransmission can result in an elevation in striatal Fos levels. This elevation may occur as a direct result of decreased dopaminergic neurotransmission or indirectly via elevated corticostriatal glutamatergic neurotransmission which occurs secondary to dopamine depletion. To test the hypothesis that elevated N-methyl-D-aspartic acid (NMDA)-mediated corticostriatal transmission may underlie the increase in striatal Fos levels upon dopamine depletion, rats were unilaterally 6 hydroxydopamine lesioned under anaesthesia induced by either barbiturate or the NMDA antagonist, ketamine. Following surgery the animals remained under light anaesthesia for 6 h prior to sacrifice and quantification of striatal Fos immunoreactivity. The results demonstrate that dopamine depletion following 6 hydroxydopamine lesioning can result in elevated striatal Fos levels which can be attenuated by contiguous treatment with an NMDA antagonist. This suggests that the increase in striatal Fos levels observed following dopamine depletion may occur as a result of elevated cytoplasmic calcium levels in the striatal cells. PMID- 7478218 TI - The effect of clozapine on Fos protein immunoreactivity in the rat forebrain is not mimicked by the addition of alpha 1-adrenergic or 5HT2 receptor blockade to haloperidol. AB - The involvement of alpha 1-adrenergic and 5HT2-receptor blockade in the induction of Fos protein produced by the 'atypical' neuroleptic clozapine was investigated in the rat forebrain. The Fos protein immunohistochemical technique has been used to identify the anatomical substrate underlying the effects of typical and atypical neuroleptics. Clozapine (20 mg/kg) induced a significantly higher Fos protein immunoreactivity response in the medial prefrontal cortex and a significantly lower response in the dorsolateral striatum compared to the effect of haloperidol (1 mg/kg). The alpha 1-adrenergic antagonist prazosin (0.3 and 1.0 mg/kg) and the 5HT2 antagonist ritanserin (1 and 3 mg/kg) did not increase Fos protein immunoreactivity by themselves and did not mimic the clozapine response when co-administered with haloperidol (1 mg/kg). Consequently, this study suggests that neither alpha 1-adrenergic receptor blockade nor the 5HT2-receptor blockade accounts for the unique Fos protein expression pattern produced by clozapine. PMID- 7478219 TI - Electrophysiological study on spinal antinociceptive interactions between adenosine and morphine in the dorsal horn of the rat. AB - The effects of intrathecal N6-cyclopentyladenosine (N6-CPA), an A1-receptor agonist, and morphine, a mu opioid-receptor agonist were studied on the electrically evoked responses of deep dorsal horn neurones in the rat. Both caused inhibitions, when applied alone, of the C-fibre evoked response and post discharges. Enhanced inhibitory effects were observed when N6-CPA (5 micrograms) was given with morphine (1 microgram). Increasing the dose of morphine (5 micrograms) with the same dose of N6-CPA occluded the expected inhibitions. By contrast, the A delta-fibre evoked response, inhibited by morphine alone, was significantly facilitated by the combination. Parallel activation of purinergic and opioid inhibitory systems within the dorsal horn as well as complex interactions between them are likely. PMID- 7478221 TI - Region-specific effects of chronic lithium administration on neuropeptide Y and somatostatin mRNA expression in the rat brain. AB - The aim of this study was to examine the effects of 4-week lithium (Li+) food supplementation on neuropeptide mRNA expression in the rat brain. In situ hybridisation was used to determine the effects on the expression of neuropeptide Y (NPY) and somatostatin (SS) mRNA. Increases in NPY mRNA levels were seen in the hippocampus, layers II-III of the entorhinal cortex, nucleus accumbens shell and in the medial caudate-putamen. Increases in SS mRNA expression were seen in the layers IV-VI of the entorhinal cortex and in the lateral caudate putamen. Thus, Li+ appears to affect discrete populations of NPY and SS mRNA expressing neurons, with possible relevance to beneficial effects of Li+ in affective disorders. PMID- 7478220 TI - 5-Hydroxytryptamine-immunoreactive nerve fibers in the rat and porcine prevertebral sympathetic ganglia: effect of precursor loading and relation to catecholaminergic neurons. AB - Localization of 5-hydroxytryptamine immunoreactivity was studied in the rat coeliac-superior mesenteric ganglion complex and in the porcine superior and inferior mesenteric ganglia by the indirect immunofluorescence technique. In normal rats, only 5-hydroxytryptamine immunoreactive SIF cells were seen in the coeliac-superior mesenteric ganglion complex. In the rats, pretreated with a 5 hydroxytryptamine precursor, L-tryptophan, and with a monoamine oxidase inhibitor, nialamide, a large number of 5-hydroxytryptamine-immunoreactive nerve fiber terminals were detected. In normal porcine superior and inferior mesenteric ganglia, intense 5-hydroxytryptamine immunoreactivity was found in numerous nerve fibers which were located around tyrosine hydroxylase-immunoreactive principal neurons. The origin and function of these fibers are discussed. PMID- 7478222 TI - Re-emergence of GAP-43 in cochlear nucleus and superior olive following cochlear ablation in the rat. AB - The effect of cochlear lesion on the expression of the growth associated protein GAP-43 in superior olive and cochlear nucleus was studied in the rat. In normal development of these auditory brainstem nuclei, GAP-43 immunoreactivity is high perinatally but low 10 days postnatally or thereafter. Removal of one spiral ganglion in grown-up animals caused a substantial re-emergence of GAP-43 immunoreactivity in varicose fibers of the ipsilateral ventral cochlear nucleus and cell bodies of the lateral superior olive. These findings suggest that a reactive synaptogenesis takes place in the cochlear nucleus as a consequence of deafening through spiral ganglion loss. PMID- 7478223 TI - Expression of Ca(2+)-ion permeable alpha-amino-3-hydroxy-5-methyl-4 isoxazolepropionate (AMPA) receptors in Xenopus oocytes injected with total RNA from human epileptic temporal lobe. AB - By using the Xenopus oocyte as an expression system, we have performed a series of experiments in order to examine the divalent cation-permeability of alpha amino-3-hydroxy-5-methyl-4-isoxazolepropionate (AMPA) receptors from the human epileptic temporal lobe. Xenopus oocytes, injected with total RNA from the epileptic temporal lobe, were tested for expression of receptors by a conventional two electrode voltage-clamp technique. Administration of glutamate and AMPA gave small or no clear current responses, whereas kainate produced large inward non-desensitizing currents. The current responses evoked by kainate were concentration dependent. Experimental data gave a Hill coefficient of 1.06 and an EC50 value of 87 microM. The current to voltage relationship showed an inward rectification and when the concentration of divalent cations were enhanced, there was a shift in the reversal potential from -11 mV (2 mM Ca2+) to 12 mV (60 mM Ba2+). This yielded a pBa2+/pK+ permeability ratio of 1.6 when the constant field equation was used. The amplitude of the currents evoked by 600 microM kainate in solutions containing higher Ba(2+)-ion concentrations was markedly diminished (46% in 10 mM Ba(2+)- and 75% in 60 mM Ba(2+)-solution), when compared to those obtained in normal Ringer's solution, suggesting interactions between different cation species and/or screening of surface charges. PMID- 7478224 TI - Biphasic edema development after experimental brain contusion in rat. AB - The time course of edema development following experimental brain contusion was studied by measuring cortex specific gravity 1 and 12 h after the trauma, and thereafter once daily until 7 days after the trauma. A biphasic development of edema was observed; the specific gravity decreased to a minimum on day 2 (P < 0.001), increased to an almost normal level on day 4 and thereafter decreased again to a second minimum 6 days after the trauma (P < 0.01). Delayed edema formation has been recognized in clinical settings, but has not been described in experimental studies. This study, with a prolonged daily follow-up, clearly demonstrates that a secondary phase of edema is an experimentally reproducible entity. The model will enable study of the pathogenetic mechanisms. PMID- 7478226 TI - Serotonin depletion by 5,7-dihydroxytryptamine does not affect G protein subunit levels in rat cortex. AB - To investigate the role of G proteins in denervation supersensitivity of the CNS serotonergic system, we examined the effect of lesioning serotonergic neurons on the abundance of cerebral cortical membrane G protein subunits in rats. Three weeks after intracisternal injection of 5,7-dihydroxytryptamine (5,7-DHT), which significantly reduced cortical 5-hydroxytryptamine (5-HT; -90%) and 5 hydroxyindoleacetic acid (approximately 98%) levels, no statistically significant differences were observed for G alpha s-1, G alpha s-s, G alpha i1, G alpha i2, G alpha q/11, G alpha 0, G beta 1 and G beta 2 immunoreactivity levels between sham lesioned and 5,7-DHT lesioned rats. These data suggest that the functional supersensitivity of 5-HT neuronal system often observed following lesions of 5-HT fibers may not involve changes at the level of G proteins but may instead encompass other downstream elements of the 5-HT receptor signaling cascade. PMID- 7478225 TI - Calpain inhibitors reduce depolarization induced loss of tau protein in primary septo-hippocampal cultures. AB - We studied the effects of a 6-min potassium depolarization injury produced by 60 mM KCl and 1.8 mM or 5.8 mM extracellular CaCl2 on tau protein levels in primary rat septo-hippocampal cultures. One day after injury, Western blot analyses revealed a calcium dependent loss of tau protein of approximately 50% of control values. Loss of tau protein was associated with calpain 1 mediated breakdown products to alpha-spectrin. Calpain inhibitors 1 and 2, applied immediately after depolarization injury and available to cultures for 24 h reduced depolarization induced degradation of tau protein to approximately 35% or 25% of control values, respectively. We propose that brief potassium depolarization causes degradation of tau protein, possibly due to calpain activation. Thus, calpain inhibitors could represent a viable strategy for preserving the cytoskeletal structure of injured neurons. PMID- 7478227 TI - Decrease in synapsin I staining in the hypogastric ganglion of aged rats. AB - The rat hypogastric ganglion (HG) contains populations of both sympathetic and parasympathetic postganglionic neurons which supply the lower pelvic viscera. These neuron populations can be identified by tyrosine hydroxylase (TH) and NADPH diaphorase (NADPH-d) staining, respectively. The effects of age on the distribution of synapsin I, a nerve terminal marker, in relation to these neuron populations has been investigated in young adult and aged rats. Most synapsin staining was axosomatic and was markedly reduced in the aged animals particularly in relation to sympathetic (NADPH-d-negative/TH-positive) neurons. Image analysis of synapsin I staining in relation to individual sympathetic neurons confirmed that there was a reduction with age of about 50% but no change in synapsin I staining in relation to parasympathetic neurons. These results suggest that synaptic transmission and peripheral integration may be affected in old age and that the autonomic control of the pelvic viscera may be compromised as a result, particularly with regard to the sympathetic innervation. Other autonomic ganglia were also studied for comparison but no such age-related differences were observed. PMID- 7478228 TI - Developmental changes in glutamate receptor stimulated inositol phospholipid metabolism and 45Ca(2+)-accumulation in posthatch chicken forebrain. AB - In chicken forebrain the two phases of synapse development, formation and maturation, are temporally well separated. We have used this model system to determine the developmental profile of glutaminergic activation of phosphoinositidase C. Stimulation of [3H]inositol-loaded forebrain prisms by quisqualic acid (QA; 30 microM), or the metabotropic agonist 1-aminocyclopentane trans-1,3-dicarboxylic acid (trans-ACPD; 30 microM), significantly increased [3H]inositol phosphate production. This response progressively decreased with developmental age, with the largest (approximately 3-fold) decrease occurring between 21 days and adult (> 10 weeks). In contrast, QA (30 microM) stimulated a quite distinct developmental profile for 45Ca2+ accumulation, with the response being maximal between 7 and 14 days before declining sharply to adult levels by 21-25 days. These results demonstrate that there is a major decrease in metabotropic glutamate receptor activation of phosphoinositidase C during the maturation phase of synapse development. PMID- 7478229 TI - Agmatine, a novel hypothalamic amine, stimulates pituitary luteinizing hormone release in vivo and hypothalamic luteinizing hormone-releasing hormone release in vitro. AB - Agmatine, a clonidine displacing substance and imidazoline receptor agonist, was recently isolated from bovine brain and shown to be present in the rat hypothalamus. Since clonidine can stimulate the release of pituitary luteinizing hormone (LH), we tested the hypothesis that agmatine may similarly act in the rat to stimulate the hypothalamic luteinizing hormone-releasing hormone (LHRH) pituitary LH axis. Administration of agmatine intracerebroventricularly rapidly augmented the release of LH in a dose-related fashion in ovariectomized, ovarian steroid-primed rats. Additionally, agmatine enhanced the in vitro efflux of LH releasing hormone from the median eminence-arcuate nucleus of the hypothalami of rats similarly pretreated with steroids. These studies imply that the endogenous imidazoline receptor agonist, agmatine, may serve as an excitatory neurotransmitter/neuromodulator in the hypothalamic control of LH release and we suggest that the previously reported excitatory effects of clonidine on LH release may be attributed to stimulation by clonidine of imidazoline receptors. PMID- 7478230 TI - Role of alpha 1 and alpha 2-adrenoceptors of the lateral hypothalamic area on urinary excretion caused by centrally administered angiotensin II. AB - To determine whether central alpha 1 and alpha 2-adrenergic mechanisms are involved in urinary sodium and potassium excretion and urine volume induced by angiotensin II (ANGII), these renal parameters were measured in volume-expanded Holtzman rats with cannulas implanted into lateral ventricle (LV) and lateral hypothalamus (LH). The injection of ANGII into LV in rats with volume expansion reduced the sodium, potassium and urine excretion in comparison to the control injections of isotonic saline, whereas prazosin (alpha 1 antagonist) potentiated these effects. Clonidine (alpha 2 agonist) and yohimbine (alpha 2 antagonist) injected into LH previous to injection of ANGII into LV also abolished the inhibitory effect of ANGII. These results suggest that the discharge of central alpha-adrenergic receptors has dual inhibitory and excitatory effect on antinatriuretic, antikaliuretic and antidiuretic effect induced by central ANGII in volume-expanded rats. PMID- 7478231 TI - Activation of the metabotropic glutamate receptor is neuroprotective during nitric oxide toxicity in primary hippocampal neurons of rats. AB - Metabotropic glutamate receptors (mGluRs) can influence neuronal survival and have been shown to be neuroprotective during glutamate toxicity in retinal cells and in cortical neurons. The mechanisms that mediate protection by this group of receptors are not clear. Since nitric oxide (NO) production can lead to neuronal cell death during excessive glutamate release, we examined whether neuronal survival was directly linked to mGluR activity and the NO pathway. Treatment with the mGluR4 receptor subtype agonist, L-(+)-2-amino-4-phosphonobutyric acid, in hippocampal cell cultures protected neurons during NO exposure. Treatment with L (+)-2-amino-3-phosphonopropionic acid, an antagonist of the mGluR1 receptor subtype and inhibitor of inositol trisphosphate formation, did not significantly alter neuronal survival during NO administration. We conclude that activation of the mGluR4 receptor protects hippocampal neurons from NO toxicity and that the mechanism of NO induced neurodegeneration does not appear to involve inhibition of the mGluR1 receptor subtype activity or the phosphoinositide system. PMID- 7478232 TI - Relationships between the force of voluntary leg movements and the associated postural adjustments. AB - Two types of kicking movements were studied: "touch mode" (minimal impact force) and "strike mode" (maximal impact force). Notable differences have been found to exist between the two kicking conditions. In the "strike mode", the onset of the voluntary movement occurred later and the movements of the postural segments preceding the voluntary movement were significantly longer and larger. These results confirm that the early postural adjustments contribute importantly to the efficiency of these movements. PMID- 7478233 TI - Up-regulation of trkB mRNA expression in the rat striatum after seizures. AB - The present study investigates the expression of a tyrosine kinase receptor (trkB), its specific ligands brain-derived neurotrophic factor (BDNF) and neurotrophin-4 (NT-4) mRNAs in the striatum after seizures. The result showed an increase of trkB mRNA expression, both with and without tyrosine kinase domain, in the caudate-putamen and nucleus accumbens, but not in the globus pallidus. This increase peaked 3 h after treatment, and returned to normal levels by 12 h. The BDNF and NT-4 mRNAs showed no change at any time. In conclusion, the widespread and massive trkB mRNA induction after abnormal neuronal activity suggests local trophic support for this receptor, and a potential role in basal ganglia diseases involving non-dopaminergic components. PMID- 7478234 TI - Tonic modulation of neurotransmitter release in the guinea-pig myenteric plexus: effect of mu and kappa opioid receptor blockade and of chronic sympathetic denervation. AB - We have studied the effects of mu- and kappa-opioid receptor blockade on endogenous acetylcholine and noradrenaline overflow from the myenteric plexus of the guinea-pig isolated colon. Cyprodime (putative mu-selective antagonist) and nor-binaltorphimine (kappa-selective antagonist) had a concentration-dependent facilitatory effect on both acetylcholine and noradrenaline overflow. Moreover, in colonic specimens obtained from sympathetically denervated animals, the effect of opioid antagonists on acetylcholine overflow was significantly higher with respect to normal preparations. Evidence is thus given in favour of an involvement of mu- and kappa-opioid receptor pathways in the tonic modulation of neurotransmitter release at the colonic level. Enhanced sensitivity to the effect of mu and kappa antagonists after chronic sympathetic denervation is strongly suggestive for the existence of a functional link between opioid and adrenergic pathways in this model. PMID- 7478235 TI - Activity-linked alterations in cholecystokininB receptor messenger RNA levels in magnocellular hypothalamic neurones by food and water deprivation in the rat. AB - CholecystokininB receptors in the hypothalamic supraoptic and paraventricular nuclei may be involved in the regulation of appetite and neuroendocrine function. In situ hybridisation was used to determine levels of mRNA encoding cholecystokininB receptors in these nuclei in normal rats and rats deprived of food, water or both food and water for 4 days. Food deprivation produced no significant change in the level of cholecystokininB mRNA in these hypothalamic nuclei. Water deprivation increased cholecystokininB mRNA levels (412 +/- 16% and 1009 +/- 69% of control for supraoptic and paraventricular nuclei, respectively) while combined food and water deprivation resulted in significantly smaller increases in these nuclei (193 +/- 20% and 303 +/- 44% of control). Increases in the paraventricular nucleus were most prominent in magnocellular (especially oxytocin-rich) subdivisions of this nucleus. These selective alterations are consistent with similar changes in cholecystokinin receptor density in these hypothalamic nuclei, and support an autocrine/paracrine role for cholecystokinin in the regulation of appetite and endocrine function via effects on hypothalamic oxytocinergic activity. PMID- 7478236 TI - Effects of long-term ovariectomy and ovarian steroids on somatogenic binding sites in rat brain and liver. AB - The effects of long-term ovariectomy and replacement with ovarian steroids on the levels of brain and liver somatogenic binding sites as well as plasma and liver growth hormone (GH) were studied in sham-operated (Sham) and ovariectomized female rats receiving either, 17 beta-estradiol (OVX-E), progesterone (OVX-P), or vehicle (OVX). Long-term ovariectomy decreased the levels of somatogenic binding sites in the choroid plexus and liver as well as GH in plasma and liver. The levels of these sites in the choroid plexus were partially restored only by estradiol replacement. Moreover, exogenous estradiol but not progesterone restored the levels of plasma and liver GH as well as liver somatogenic binding sites. Our results suggest that estrogens regulate the levels of somatogenic binding sites in the liver and choroid plexus. PMID- 7478237 TI - Effect of a neuron-specific actin-binding protein, drebrin A, on cell-substratum adhesion. AB - Drebrin A expression was induced in non-neuronal L cells via transfection with a vector containing the cDNA of rat drebrin A. Following treatment with colcemid (5 micrograms/ml) and cytochalasin D (0.5 micrograms/ml), most L cells collapsed into round cells, while drebrin expressing cells were resistant to the treatment, keeping their cell shapes. Simultaneously, actin filaments and microtubules were disrupted in both cell lines. By quantitative analysis, in the presence of cytochalasin D, the extent of cell spreading and cell attachment in drebrin expressing cells was significantly higher than that in control cells. These results suggest that drebrin A modulates cell-substratum adhesion. PMID- 7478238 TI - Seizure-induced c-fos expression in rat medulla oblongata is not dependent on associated elevation of blood pressure. AB - This study was performed to determine whether c-fos immunoreactivity (IR) induced in medulla oblongata by pentylenetetrazole seizures is a consequence of seizure associated blood pressure elevation and activation of baroreceptor afferent pathways, or occurs independently of hypertension. Immunohistochemical study of sections of medulla oblongata revealed that seizures are followed by induction of c-fos IR in nucleus tractus solitarius (NTS), dorsal motor nucleus of the vagus (DMN 10), and ventrolateral medulla (VLM), while there is negligible c-fos IR after saline sham injections. Seizures were associated with blood pressure elevation peaking at 31 +/- 17% (+/- SD) above baseline. Experimental hypertension at a similar level induced by i.p. phenylephrine also resulted in induction of c-fos IR in NTS. When seizures were preceded by antihypertensive treatment with the alpha-adrenergic antagonist, phentolamine, peak blood pressure tended to remain near the baseline level and lower than sham-injected controls. Normotensive seizures were associated with c-fos IR in NTS, DMN 10, and VLM similar to the pattern following hypertensive seizures. Seizure-induced activation of c-fos IR occurred despite normal blood pressure, and thus can be attributed to a direct effect of the seizure, and not to an indirect effect mediated by hypertension. PMID- 7478239 TI - Increased Cu/Zn superoxide dismutase-like immunoreactivity in the swollen axons of rats intoxicated chronically with beta,beta'-iminodipropionitrile. AB - Demonstration of a genetic linkage between the Cu/Zn superoxide dismutase (SOD1) gene and familial amyotrophic lateral sclerosis (ALS) has aroused interest in the role of SOD1 in spinal motoneuronal death. We used chronically beta,beta' iminodipropionitrile (IDPN)-intoxicated rats as a model of ALS and investigated SOD1 changes in the spinal cord by immunocytochemical and in situ hybridization techniques. Compared with control rats, SOD1-like immunoreactivity (SOD1-IR) increased in swollen axons of the proximal spinal roots, but not in motoneuronal and dorsal root ganglion neuronal cell bodies where SOD1 gene transcription did not increase. The present data indicate that treatment with IDPN induces accumulation of SOD1 in the swollen axons by blocking slow axonal flow, suggesting the possibility that increased SOD1-IR in ALS is induced by axonal flow blockade. PMID- 7478240 TI - Modulation of N-methyl-D-aspartic acid receptors by extracellular calcium in immature and adult hippocampal slices: whole cell recordings in CA3 pyramidal cells. AB - Lowering extracellular calcium concentration [Ca2+]o in rat hippocampal slices can lead to an induction of epileptiform activity. It has been shown that this effect is more pronounced in slices of neonatal rats (postnatal day, PND 8-19) than in mature slices (> PND 40) and it has been suggested that unique N-methyl-D aspartic acid (NMDA) receptor properties of immature rat hippocampal pyramidal cells contribute to this developmental effect. In a voltage clamp experiment we tested NMDA receptor properties in hippocampal pyramidal cells by measuring NMDA receptor mediated currents evoked by iontophoretic applied NMDA in the basal dendrites of CA3 pyramidal neurons. We found that lowering extracellular calcium from 2 to 1 mM, increases NMDA evoked inward current in pyramidal cells around the resting membrane potential. However, this effect is observed in slices of neonatal as well as in slices of mature rats, suggesting that there is no difference in NMDA receptor sensitivity to extracellular Ca2+ between these two age groups. The modulation of the NMDA receptor by extracellular calcium at physiological concentrations can have important consequences in pathological conditions during which extracellular calcium reaches low levels. Because this 'hypocalcemic' condition induces a larger current influx via the NMDA receptor channel at resting membrane potentials, it can further enhance cellular excitability and contribute to sustain epileptiform activity. PMID- 7478241 TI - Induction of nitric oxide synthase activity in rodent brain following middle cerebral artery occlusion. AB - The conversion of L-[3H]arginine to L-[3H]citrulline in the absence of calcium can be used to assay selectively the activity of inducible nitric oxide synthase (NOS) in rat spleen homogenates 6 h after lipopolysaccharide administration. Using similar assay conditions, changes in inducible NOS activity were measured within ischemic brain tissue between 2 h and 7 days following permanent middle cerebral artery (MCA) occlusion in Sprague-Dawley rats and SV-129 mice. Total (constitutive and inducible) NOS activity was measured in the presence of 0.5 mM CaCl2. Whereas total NOS activity in rat decreased dramatically to 16% and 6% of baseline 6 and 12 h after MCA occlusion, inducible NOS activity remained undetectable before 2 days after occlusion, became maximal at 3 days, and decreased to less than 10% of maximal iNOS activity at 7 days. In the mouse, total NOS activity decreased after MCA occlusion but inducible NOS activity was undetectable from 2 h to 4 days after occlusion. Sustained NO production by inducible NOS activity does not contribute to ischemic injury within 24 h after MCA occlusion, but may contribute to infarct maturation 2-4 days after ischemia in some but not all species. PMID- 7478242 TI - Altered expression of dopamine D2 receptor mRNA splice variants in brain and pituitary of spontaneously hypertensive rats. AB - Both central and peripheral dopamine (DA) has been shown to play a role in the regulation of blood pressure. Using sensitive nuclease protection analysis, we have compared the expression of DA D-2 receptor (D2-R) mRNA splice variants in brain and pituitary of spontaneously hypertensive rats (SHR) with normotensive Wistar-Kyoto (WKY) controls. Levels of D2-R mRNA were significantly altered in pituitary anterior lobe (AL) and neurointermediate lobe (NIL), and in striatum of SHR, but not in any other brain regions examined. SHR pituitary expressed 50-80% higher levels of D2-R mRNA, coupled with an increase in the relative proportion of the long (D2-L) mRNA variant. In contrast, overall D2-R mRNA expression in SHR striatum was only 75% that of WKY controls, however, the relative proportion of the D2-L splice variant was increased. The present data demonstrate that tissue specific alterations in D2-R mRNA levels and primary transcript splicing exist in the SHR and suggest that these changes may in part mediate differential responsiveness to DA that may be related to the development of hypertension. PMID- 7478243 TI - Vigabatrin protects against kainic acid-induced neuronal damage in the rat hippocampus. AB - We studied the neuroprotective effect of vigabatrin (gamma-vinyl GABA, VGB) in the rat hippocampus after status epilepticus (SE) induced by kainic acid (KA). Rats were treated with VGB (500 or 1000 mg/kg, i.p.) 24 h before KA injection (9 mg/kg, i.p.). The lower dose of VGB had no effect on the generation or severity of convulsions. However, VGB decreased neuronal damage in the CA3a (P < 0.05) and CA1 (P < 0.01) subfields of the hippocampus. The higher dose of VGB attenuated the severity of convulsions (P < 0.05) but had no effect on the development or generalization of convulsions. This finding may have clinical implications in the prevention of neuronal damage induced by drug refractory seizures or SE. PMID- 7478244 TI - Evidence for soleus H-reflex depression induced by ballistic head rotation. AB - In this study, we described a sequence of changes of the electromyographic (EMG) and depression of the soleus (Sol) H-reflex that precede and follow the onset of ballistic head rotation in freely standing human subjects. During head rotation, a sequence of EMG changes occurred simultaneously in Sol and in biceps femoris muscle (BF). Specifically, changes to Sol EMG included deactivations (EMG silent period) and H-reflex depression coincident with BF excitation. Thus, in the case of ballistic head movement, control commands for anticipatory postural adjustment appear to excite BF and inhibit Sol in a similar way to that previously described for the onset of arm movement. PMID- 7478247 TI - Somatosensory evoked magnetic fields after mechanical stimulation of the scalp in humans. AB - Somatosensory evoked magnetic fields after mechanical stimulation by air-pressure induced tapping which was applied to the forehead and occiput were examined in normal human subjects. The equivalent current dipole (ECD) of the initial magnetic field, 1M, was identified in the primary somatosensory cortex (SI) in the hemisphere contralateral to the stimulation. The ECD of the subsequent magnetic fields, 2M, was identified in bilateral second sensory cortices (SII). The ECD position of 1M in SI generated after the scalp stimulation was closely inferior to the hand area of the SI, which was consistent with the well-known somatotopic organization, 'homunculus'. PMID- 7478245 TI - Functional assessment of Ca(2+)-current in the mouse motor nerve terminals. AB - The purpose of this study was to characterize voltage-gated Ca2+ channels on the mouse motor nerve terminals. Mouse diaphragm and triangularis sterni preparations were used for this study in order to assess the functional Ca2+ channels in the transmitter release. The results showed that omega-conotoxin MVIIC (CTx-MVIIC, 0.5-1 microM) but not omega-conotoxin GVIA (1 mM) markedly inhibits not only the nerve-evoked muscle contractions accompanied by a decrease in the amplitude of end plate potentials (epps) in the mouse phrenic-nerve diaphragm but also the Ca(2+)-waveforms in the nerve terminals of triangularis sterni. The inhibitory effects of CTx-MVIIC were considered to be specifically presynaptic rather than myogenic, since none of the electrical properties of muscle fibers including action potentials, resting membrane potentials and the miniature endplate potential, were affected. Moreover, Na(+)- and K(+)-waveforms of the nerve terminals were unaffected by CTx-MVIIC. At a saturating concentration of 3-5 mM, CTx-MVIIC exerted a maximal inhibitory effect by 38% of 3,4-diaminopyridine prolonged epps area and inhibited only the slow component of Ca(2+)-current, respectively, and the remaining fast component could be inhibited by subsequent addition of cadmium chloride (Cd2+). All of these findings indicate that at least two components (a slow CTx-MVIIC sensitive component and a fast Cd2+ sensitive component) of the mouse motor nerve terminals would cooperate in the induction of the transmitter release from motor nerve endings. PMID- 7478246 TI - The role of cerebral microvessels in the elimination of histamine released during postasphyxial reperfusion in newborn piglets. AB - Histamine, released from intracerebral sources during hypoxic-ischemic conditions, may take part in the pathogenesis of neonatal brain injuries. In order to elucidate the possible role of cerebral microvessels in the elimination of histamine from the extracellular space, we determined the concentration of histamine using a modified radioenzymatic method in plasma taken from the internal jugular vein, in cerebrospinal fluid, and in capillary-rich fraction of cerebral microvessels prepared from cortex in 12 sham-operated piglets. Then, bilateral pneumothorax was induced in 20 piglets, samples were taken from the same compartments as from the controls before and during asphyxia, as well as 15 and 180 min thereafter, respectively. Plasma histamine level was significantly (P < 0.05) elevated in animals during hypoxic cardiovascular and metabolic failure (13.5 +/- 1.9 nM l-1) compared to value measured in the control group (2.2 +/- 0.5 nM l-1), preceding any detectable change of histamine concentration in cerebrospinal fluid (5.2 +/- 1.9 versus 3.8 +/- 1.1 nM l-1, respectively) or in cerebral microvessels (8.4 +/- 0.8 versus 7.1 +/- 0.6 pM (mg protein)-1). After resuscitation, histamine levels in plasma samples remained high during the early (15 min, 16.2 +/- 4.3 nM x l-1) and late (180 min, 15.3 +/- 2.9 nM l-1) reperfusion period. By contrast, histamine concentration was increased considerably (P < 0.05) in cerebrospinal fluid samples obtained 15 min (12.8 +/- 6.5 nM l-1), but not 180 min (5.2 +/- 1.9 nM l-1) after resuscitation.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7478248 TI - Intrathecal injection of an oxytocin-receptor antagonist attenuated postnephrectomy natriuresis in the male rat. AB - We have previously shown that oxytocin (OT) is a major humoral mediator in postnephrectomy natriuresis. As immunoassayable OT has been demonstrated in the spinal cord, the aim of this investigation was to determine whether OT receptors in the spinal cord are also involved in this natriuresis. The experiments were performed on anesthetized male rats. Before acute unilateral nephrectomy, an oxytocin-receptor antagonist was injected intrathecally in the thoracolumbar region in rats. Postnephrectomy natriuresis was attenuated by this injection but not by intrathecal injection of artificial cerebrospinal fluid. Our results suggest that OT receptors within the spinal cord may influence the autonomic nervous regulation of renal function. In an additional experiment, intravenously infused hexamethonium did not prevent the adaptive natriuresis in the remaining kidney. We conclude that OT receptors in the spinal cord are involved in the postnephrectomy natriuresis, possibly as a component in the afferent signal pathway. PMID- 7478249 TI - Regulatory mechanisms involved in the activation of bradykinin-induced membrane currents in PC12 cells. AB - Whole-cell patch-clamp measurements were made in nerve-growth-factor (NGF) treated PC12 cells. External application of bradykinin (BK) activated an outward and an inward current which could be separated by using KCl- or CsCl-containing pipette solutions. The slowly activating inward current could be induced by BK independently of the filling of intracellular Ca2+ stores. By using GDP-beta-S in the pipette medium, we showed that BK-induced outward and inward currents were differentially regulated through G-protein-sensitive and -insensitive mechanisms, respectively. While the outward current was inhibited by GDP-beta-S, the inward current was not affected. Our results show that occupancy of BK receptors activates different signaling pathways for the induction of outward and inward currents. PMID- 7478250 TI - Further observations on MxA-positive Lewy bodies in Parkinson's disease brain tissues. AB - MxA protein, one of the GTPases, is induced in the cytoplasm by interferons and confers a high degree of resistance to some viruses. We recently reported that in cases of Parkinson's disease (PD), Lewy bodies (LBs) in the substantia nigra (SN) were positive for MxA. This study aimed to provide more detailed data on MxA positive structures in the SN in PD brain tissues. Swellings of neuronal processes, as well as LBs, were positive for MxA. Immuno-electron microscopy revealed that reaction products corresponding to MxA proteins localized in self aggregations similar to those reported for Mx1 protein in mouse tissues. These results show that MxA may play a role in the formation of LBs and swellings of neuronal processes. PMID- 7478251 TI - Inhibition of inducible nitric oxide synthase mRNA expression by basic fibroblast growth factor in human microglial cells. AB - The effect of basic fibroblast growth factor (bFGF) on inducible nitric oxide synthase (iNOS) mRNA expression in human cultured ramified microglial cells was investigated. Using RT-PCR and Southern blot analysis, we found that bFGF prevented the iNOS gene expression as induced by LPS/TNF alpha. Also, bFGF dose dependently inhibited nitrite levels in treated cell supernatants. That the early presence of bFGF during LPS/TNF alpha induction was essential indicates that iNOS gene expression can be transcriptionally regulated. PMID- 7478252 TI - Presence of estrogen receptors in aromatase-immunoreactive neurons in the mouse brain. AB - The present study was undertaken to demonstrate estrogen receptors in aromatase immunoreactive neurons showed the immunoreactivity for estrogen receptors in the cell nuclei in the medial subdivision of the bed nucleus of the stria terminalis and the posterodorsal division of the medial amygdaloid nucleus, but a few did in the medial preoptic area. These results suggest that aromatase-immunoreactive neurons in the former two areas are regulated by steroids through estrogen receptors in their cell nuclei. PMID- 7478253 TI - ApoE immunoreactivity and microglial cells in Alzheimer's disease brain. AB - The spatial relationship of apolipoprotein E (apoE)-like immunoreactivity (IR) to amyloid beta-peptide (A beta), astrocytes and microglial cells in the brain of Alzheimer's disease was studied by double immunolabelling. Diffuse apoE-like IR was seen in A beta diffuse deposits, and markedly increased in the core of classic senile plaques. Microglial cells, sometimes immunoreactive for apoE, were frequent in areas of apoE-like IR, where they often grouped into clusters in the core of apoE-labelled senile plaques. Although astrocytic processes were seen within these senile plaques, the cell bodies were always at a distance from the core. None of these astrocytes expressed apoE-like IR. Microglial cells, some of them immunoreactive for apoE, were seen in the center of apoE-labelled senile plaques. These data suggest that microglial cells play a more significant role than astrocytes in apoE deposition in senile plaques of Alzheimer disease. PMID- 7478254 TI - Tetrahydroaminoacridine modulates technetium-99m labelled ethylene dicysteinate retention in Alzheimer's disease measured with single photon emission computed tomography imaging. AB - The present study investigated if acute treatment with tetrahydroaminoacridine (THA) (25 or 75 mg, p.o.) affects technetium-99m labelled ethylene dicysteinate (ECD) retention abnormalities in patients with mild to moderate Alzheimer's disease (AD; mean age 69 years). THA (75 mg) increased temporal, prefrontal and occipital ECD retention (normalized to cerebellum) in mildly demented AD patients, but 25 mg of THA had no effect on ECD retention. After 75 mg THA, prefrontal and temporal ECD retention correlated with improved executive and memory functioning, respectively. THA (25 or 75 mg) had no measurable effect on ECD retention of moderately demented patients. PMID- 7478256 TI - Response characteristics of cutaneous mechanoreceptors to vibratory stimuli in human glabrous skin. AB - We investigated the responsiveness of mechanoreceptors in the glabrous skin of the hand to triangular vibratory stimuli when the frequency and applied pressure to the skin were varied. Using a microneurographic technique, 13 single afferent activities were recorded from 4 kinds of mechanoreceptors. Both fast-adapting (FA) and slow-adapting (SA) units were sensitive to the vibratory stimuli. The relationship between the most sensitive frequency and applied pressure to the skin were analyzed as a tuning curve. FA-type I (FAI) was sensitive to vibratory stimuli at 30-40 Hz and the frequency which entrained one-to-one discharge at lower pressure was between 10 and 80 Hz. FA-type II (FAII) sensitivity was augmented sharply over 60-100 Hz. SA-type I (SAI) and SA-type II (SAII) responsiveness was almost the same and characteristic sensitivity to the vibratory stimuli was revealed under 15 Hz. Functional roles of these units, especially type I units, are discussed in relation to active touch such as texture discrimination with a finger. PMID- 7478255 TI - Expression of type I interleukin-1 receptor mRNA and its regulation in cultured astrocytes. AB - The expression of type I interleukin-1 receptor (IL-1RI) mRNA enriched astrocytes and microglia was investigated by Northern blot analysis. IL-1RI mRNA was expressed in untreated astrocytes, but not in microglia. This expression was markedly suppressed by a treatment with isoproterenol, a beta-adrenoceptor agonist (1 x 10(-8) to 1 x 10(-6) M). This suppression was clearly reversed by propranolol, a beta-adrenoceptor antagonist. Dibutyryl cAMP (dbcAMP) also suppressed the expression of IL-1RI mRNA. The suppression by isoproterenol or dbcAMP was blocked by H8 or H89, inhibitors of cAMP-dependent protein kinase. The present results suggest that IL-1RI mRNA expression in cultured astrocytes may be regulated by intracellular cAMP, elevated upon the activation of beta adrenoceptors as an example, and cAMP-dependent protein kinase. PMID- 7478257 TI - Ligand-binding properties of an unusual nicotinic acetylcholine receptor subtype on isolated outer hair cells from guinea pig cochlea. AB - Acetylcholine receptors on isolated guinea pig cochlear outer hair cells (OHC) were characterized by radioligand binding. Equilibrium binding of [125I]alpha bungarotoxin revealed a KD of 62 +/- 2 nM, Bmax = 7.2 +/- 1.8 x 10(7) binding sites/OHC, and a slowly reversible dissociation rate constant, kappa-1 = 2.2 +/- 0.01 x 10(-4) min-1. L-[3H]Nicotine bound reversibly (estimated KD approximately 230 nM and Bmax approximately 5 x 10(7)) with kinetic rate constants of association kappa-1 = 6.2 +/- 0.06 x 10(4) min-1 nM-1 and dissociation kappa-1 = 0.23 +/- 0.003 min-1. [3H]Strychnine bound to OHC with a KD of 35 +/- 6 nM and Bmax = 2.6 +/- 0.5 x 10(7), and binding increased 3-4 fold after membrane depolarization with 56.2 mM [K+], suggesting additional binding sites. Binding, seen only at > nM concentrations, of [3H]3-quinuclidinyl benzilate (KD = 11.5 +/- 5 nM; Bmax = 2.5 +/- 0.6 x 10(6)) was competitively inhibited by the muscarinic antagonists atropine and 4-DAMP (IC50 of 6.1 +/- 0.5 and 6.5 +/- 0.4 nM). The OHC receptor is thus an atypical nicotinic acetylcholine receptor subtype with unusual pharmacological properties. PMID- 7478258 TI - The superior colliculus relays signals descending from the vibrissal motor cortex to the facial nerve nucleus in the rat. AB - Electrical stimulation of the vibrissal motor cortex (VMCx), the superior colliculus and the facial nerve nucleus elicits vibrissal movements. A possibility of the superior colliculus as one of the relay nuclei between the VMCx and the facial nerve nucleus was investigated by injecting an anterograde neural tracer, Phaseolus vulgaris leucoagglutinin (PHA-L), into the physiologically identified VMCx and a retrograde tracer, cholera toxin B subunit (CTb), into the facial nerve nucleus in Wistar rats. In the lateral half of the superior colliculus, the termination field of axons from the VMCx overlapped with the distribution field of projection neurons to the facial nerve nucleus. In some cases, PHA-L-labeled terminal swellings were closely apposed to the somas of CTb labeled neurons. We conclude that signals descending from the VMCx are relayed through the lateral half of the superior colliculus to the facial nerve nucleus. PMID- 7478259 TI - Methamphetamine and dopamine neurotoxicity: differential effects of agents interfering with glutamatergic transmission. AB - The effects of riluzole and lamotrigine, two agents which interfere with the release of glutamate (GLU), and MK-801, a blocker of N-methyl-D-aspartate (NMDA) receptors, were compared in the model of methamphetamine-induced depletion of dopamine (DA) levels in mice. Repeated injections with methamphetamine (4 x 5 mg/kg i.p.) markedly decreased levels of DA, 3,4-dihydroxyphenylacetic acid (DOPAC) and homovanillic acid (HVA) levels. When mice were treated with riluzole (2 x 10 mg/kg p.o.), no protection was observed against the decrease in DA and the two metabolites. Lamotrigine (2 x 10 mg/kg p.o.) was also inactive. Treatment with MK-801 (2 x 2.5 mg/kg i.p.) antagonized the decrease in DA, DOPAC and HVA levels induced by the neurotoxin. Thus, unlike an NMDA blocker, drugs that interfere with GLU release did not antagonize the methamphetamine-induced DA neurotoxicity in mice. The consequences of this inactivity are discussed in terms of the reliability of this model to test new drugs with putative efficacy in the treatment of Parkinson's disease. PMID- 7478260 TI - Monomers and dimers of acetylcholinesterase in human meningioma are anchored to the membrane by glycosylphosphatidylinositol. AB - Amphiphilic monomers and dimers of acetylcholinesterase (AChE) and hydrophilic tetramers of butyrylcholinesterase (BuChE) were released by extracting human meningioma with Tris-saline and Tris-saline-Triton X-100 buffers. The amphiphilic or hydrophilic behavior of the AChE and BuChE forms was assessed by sedimentation analysis, hydrophobic chromatography and Triton X-114 phase-partitioning. A significant fraction of the amphiphilic AChE species was converted into hydrophilic components by incubation of the soluble enzyme with phosphatidylinositol-specific phospholipase C (PIPLC) from Bacillus thuringiensis, this fraction being increased by a double treatment with PIPLC and alkaline hydroxylamine. A significant amount of the membrane-bound AChE was released by incubation with PIPLC. These results demonstrate that AChE forms in meningioma are attached to the membrane via glycosylphosphatidylinositol, although part of the enzyme forms are resistant to PIPLC. PMID- 7478261 TI - Establishment of human microglial cell lines after transfection of primary cultures of embryonic microglial cells with the SV40 large T antigen. AB - Four continuous cell lines of human microglial cells were obtained by transfection of enriched cultures of human embryonic brain-derived macrophages with a plasmid encoding for the large T antigen of SV40. The transformed cells had the macrophagic characteristics of adherence and intra-cytoplasmic non specific esterase activity. They could phagocytize zymosan particles but the phagocytic activity remained low. They expressed several macrophagic antigens but not the monocytic markers CD14, CD4, CD68/Ki-M6 and CD11c. The cells could be activated to express class II major histocompatibility complex antigens after interferon-gamma activation. Finally, interleukin-6 was produced spontaneously by the cells and this production was further increased after interleukin-1 alpha stimulation. PMID- 7478262 TI - Operantly conditioned plasticity and circadian rhythm in rat H-reflex are independent phenomena. AB - Recent studies indicate that rats can increase or decrease H-reflex amplitude in response to an operant conditioning paradigm. In addition, rats also display a circadian rhythm in H-reflex amplitude. As part of the development of H-reflex conditioning in the rat as a new model for defining the plasticity underlying a simple form of learning, this study examined the relationship in the rat between operantly conditioned H-reflex change and the H-reflex circadian rhythm. When H reflex amplitude increased or decreased in response to the operant conditioning program, its circadian rhythm showed no changes in phase and minimal change in amplitude. Furthermore, animals did not alter daily performance schedule so as to use the rhythm to increase reward probability. Thus, in the rat, H-reflex operant conditioning and the H-reflex circadian rhythm appear to be independent phenomena. The circadian rhythm should not be a significant complicating factor in studies of operantly conditioned H-reflex change. PMID- 7478263 TI - Lack of evidence for dopamine autoreceptors in the mediobasal hypothalamus: a microdialysis study in awake rats. AB - To determine the functional presence of dopamine (DA) autoreceptors on tuberoinfundibular dopamine (TIDA)-ergic neurons in awake rats, a microdialysis probe was implanted into the mediobasal hypothalamus (MBH). In the presence of the re-uptake inhibitor nomifensine, which increased DA levels to 350% of basal values, systemic administration of the non-selective D1/D2 antagonist haloperiol induced an immediate increase in DA and DOPAC levels to 145% of pretreatment values. However, neither local infusion of the selective D2 antagonist sulpiride nor of the D1 antagonist SCH 23390 affected the nomifensine-elevated extracellular DA or DOPAC levels in the MBH. Systemic administration of the D2 antagonist raclopride equally did not affect the nomifensine-elevated DA release in the MBH. Upon basal extracellular DA levels (without nomifensine), local infusion of the D2 agonist (-)N-0437 equally did not affect the DA or DOPAC levels in the MBH. Furthermore, the increase in DA levels induced by haloperidol could not be antagonized by the D1 agonist CY 208-243. Therefore, the present study does not provide support for the concept of functional autoreceptors located on TIDA neurons regulating the release of DA. Possibly, the effect of haloperidol was non-DA-ergic in character. PMID- 7478265 TI - Is the clustering of beta-amyloid (A beta) deposits in the frontal cortex of Alzheimer patients determined by blood vessels? AB - The clustering pattern of diffuse, primitive and classic beta-amyloid (A beta) deposits was studied in the upper laminae of the frontal cortex of 9 patients with sporadic Alzheimer's disease (AD). A beta stained tissue was counterstained with collagen type IV antiserum to determine whether the clusters of A beta deposits were related to blood vessels. In all patients, A beta deposits and blood vessels were clustered, with in many patients, a regular periodicity of clusters along the cortex parallel to the pia. The classic A beta deposit clusters coincided with those of the larger blood vessels in all patients and with clusters of smaller blood vessels in 4 patients. Diffuse deposit clusters were related to blood vessels in 3 patients. Primitive deposit clusters were either unrelated to or negatively correlated with the blood vessels in six patients. Hence, A beta deposit subtypes differ in their relationship to blood vessels. The data suggest a direct and specific role for the larger blood vessels in the formation of amyloid cores in AD. PMID- 7478264 TI - Identification of the Ulex europaeus agglutinin-I-binding protein as a unique glycoform of the neural cell adhesion molecule in the olfactory sensory axons of adults rats. AB - Histochemical localization of two lectins, Ulex europaeus agglutinin-I (UEA-I) and Tetragonolobus purpureus (TPA), was studied in the olfactory bulb of adult rats. In contrast to TPA, UEA-I detected a fucosylated glycoprotein that is only present in the surface membranes of olfactory sensory cells including the whole course of their neurites up to the final arborization in glomeruli. Immunoblotting revealed that UEA-I binds specifically to a protein of 205 kDa, while TPA stains several other glycoproteins. Affinity chromatography with the use of a UEA-I column identified the 205 kDa protein as a glycoform of neural cell adhesion molecule (N-CAM), specific for the rat olfactory sensory nerves. PMID- 7478266 TI - Cellular distribution of the mRNA for the kappa-opioid receptor in the human neocortex: a non-isotopic in situ hybridization study. AB - Opioid receptors (OR) provide primary interaction sites of the human brain with opiates. Presently kappa-OR mRNA expression was studied in different cortical areas (A4, A10, A17) by in situ hybridization using digoxigenin-labeled oligonucleotides and an alkaline phosphatase-mediated color reaction. kappa-OR mRNA was expressed mainly in layers II/III and V pyramidal and layer VI multiform neurons. A4 giant pyramidal and A17 giant stellate neurons stood out labeled. These findings fit in with our data on kappa-OR protein distribution. Combined cellular assessment of protein and mRNA will enable the study kappa-OR expression under physiological and pathological conditions. PMID- 7478267 TI - Survival of neurons containing the enzyme nicotinamide adenine dinucleotide phosphate (NADPH) diaphorase in static slice cultures of adult rat striatum. AB - The present study concerns the survival potential of mature neurons containing the enzyme nicotinamide adenine dinucleotide phosphate (NADPH) diaphorase in the static slice culture of adult rat striatum. In the striatal tissues immediately after slicing, there was a scattered distribution of NADPH-diaphorase neurons stained in a Golgi-like manner, and the cell density of those neurons was 53 +/- 5 (mean +/- SEM; n = 10) cells per mm2. The time-sequential cell density analysis disclosed that the number of striatal NADPH-diaphorase neurons surviving after 1, 2, 4 and 6 day in culture were 26 +/- 5, 8 +/- 2, 5 +/- 2, and 3 +/- 2 (means +/- SEM; n = 10) cells per mm2, respectively. Thus, approximately 50% of striatal NADPH-diaphorase neurons survived for 1 day and a significant proportion of these neurons, although their number gradually decreased, were maintained in culture for at least several days. The conspicuous survival of the striatal NADPH diaphorase neurons in slice culture is thought to reflect the damage-resistant natures of these cells. PMID- 7478268 TI - Vasoactive intestinal peptide but not galanin promotes survival of neonatal rat sympathetic neurons and neurite outgrowth of PC12 cells. AB - The synthesis of the neuropeptide galanin (GAL) is greatly enhanced after axonal lesion in different neuron populations of the peripheral and central nervous system. In sympathetic ganglia, GAL-immunoreactive nerve fiber baskets have been found surrounding postganglionic neurons after axotomy. Until now, it is unclear if GAL may be involved in neuronal survival or regeneration as suggested for vasoactive intestinal peptide (VIP) that is also upregulated after nerve lesion. We have, therefore, studied the effects of GAL on survival of sympathetic neurons dissociated from newborn rat superior cervical ganglia and on neurite outgrowth of PC12 cells. These effects were compared to those elicited by VIP. Whereas VIP promoted survival of about 10% of sympathetic neurons 2 days after nerve growth factor deprivation and induced neurite outgrowth of PC12 cells already at 6 h after addition of the peptide, GAL had no effect in either of these culture systems. While the induction of VIP may be beneficial for axotomized neurons, the functional significance of increased GAL levels remains to be established. PMID- 7478269 TI - Cerebrospinal fluid and serum concentrations of the N-methyl-D-aspartate (NMDA) receptor antagonist memantine in man. AB - Memantine is an uncompetitive N-methyl-D-aspartate (NMDA) receptor antagonist with therapeutic potential in dementia, spasticity and Parkinson's disease. The Ki-value of memantine at the phencyclidine (PCP) binding site of the NMDA receptor is 0.5 microM in human frontal cortex. We investigated whether concentrations of memantine in cerebrospinal fluid (CSF) and serum samples under therapeutic conditions are in the range of its Ki-value at the PCP binding site. The serum levels ranged from 0.025 to 0.529 microM with daily doses between 5 and 30 mg. CSF levels were highly correlated to serum levels and were below serum levels in each patient with a mean CSF/serum ratio of 0.52. Serum and CSF levels were correlated to the daily dose, but not to the duration of treatment. At the concentrations reported here, memantine is expected to specifically interact with the PCP binding site of the NMDA receptor. PMID- 7478270 TI - Anatomical evidence for inputs to ventrolateral medullary catecholaminergic neurons from the midbrain periaqueductal gray of the rat. AB - Previous studies have shown that the midbrain periaqueductal gray (PAG) projects to the ventrolateral medulla (VLM). Here, we studied PAG projections to the area of A1/C1 neurons in the VLM in the rat using phaseolus vulgaris leucoagglutinin (PHA-L) anterograde tracing combined with immunocytochemistry for tyrosine hydroxylase (TH) or phenylethanolamine N-methyl transferase (PNMT). Following PAG injections, PHA-L labeled fibers and terminals were intermingled among TH immunoreactive (TH-ir) neurons in the VLM. High-power light microscopic examination revealed that some of the PHA-L labeled varicose fibers and boutons were in close contiguity with TH-ir elements. Such apparent appositions appeared more frequently on TH-ir elements in the A1 area than on TH-ir or PNMT-ir neurons in the C1 area. These results indicate that some PAG inputs to the VLM may directly innervate A1/C1 neurons. PMID- 7478271 TI - Allopregnanolone potentiates a GABA-withdrawal syndrome in the rat cerebral cortex. AB - We have studied the neuromodulatory effect of the neurosteroid 3 alpha-hydroxy-5 alpha-pregnan-20-one (allopregnanolone-3 alpha-5 alpha P-) in the GABA-withdrawal syndrome (GWS). This is a model of partial epilepsy consisting of an enduring paraoxysmal activity recorded at the site of GABA infusion that depends, for its induction, on GABA receptor activation. Rats were chronically implanted for frontal and occipital EEG recording with infusion cannulae fixed on the somatomotor cortical region. When the neurosteroid was infused after or concurrently with GABA, a potentiation of the GWS (i.e. shorter latency and prolonged duration) was observed. No modifications in EEG activity were detected when allopregnanolone was administered alone or prior to GABA administration. These results indicate a neuromodulatory effect of allopregnanolone, dependent on the presence of GABA at the receptor site. PMID- 7478272 TI - Anticonvulsant properties of D-20443 in genetically epilepsy-prone rats: prediction of clinical response. AB - D-20443 is an experimental antiepileptic drug. Its mechanism of antiepileptic action is unknown. We evaluated the anticonvulsant effectiveness of D-20443 against sound-induced seizures in genetically epilepsy-prone rats (GEPRs). This compound produced anticonvulsant effects against sound-induced seizures in moderate seizure GEPRs (GEPR-3s) at significantly lower doses than in severe seizure GEPRs (GEPR-9s). Based on these data and on the responses of GEPRs to other antiepileptic drugs, we predict that D-20443 will be a broad spectrum antiepileptic agent in humans. That is, we predict that D-20443 will suppress both tonic/clonic and absence seizures in humans. PMID- 7478275 TI - Free radical formation in autopsy samples of Alzheimer and control cortex. AB - The formation of free radicals in homogenates of frontal cortex from brains taken at autopsy and verified histologically to be from five patients with Alzheimer's disease or from six age matched normal controls, was measured by electron paramagnetic resonance spectroscopy. During incubation at 37 degrees C, the formation of free radicals by Alzheimer's samples was 22% higher (P < 0.05) than controls. Following incubation in the presence of ferrous sulfate (200 microM), samples of Alzheimer's frontal cortex produced nearly 50% more free radicals than did controls (P < 0.01). Although these post mortem in vitro observations are consistent with an increased free radical burden in tissue from patients with Alzheimer's disease, that comparable differences exist in vivo between Alzheimer's patients and non-demented people remains to be demonstrated. PMID- 7478273 TI - Apoptosis and cell proliferation in human neuroepithelial tumors. AB - Apoptosis and cell proliferation were studied in 180 human neuroepithelial tumors (30 medulloblastomas, 30 intracranial ependymomas, 30 oligodendrogliomas and 90 astrocytic tumors, including 30 astrocytomas, 30 anaplastic astrocytomas and 30 glioblastomas). Apoptotic nuclei were detected by morphology and in situ end labeling (ISEL) of DNA breaks. The frequency of apoptotic nuclei varied from oncotype to oncotype and their distribution in each oncotype was uneven. An apoptotic index (AI) was calculated; this was high in malignant tumors and in tumors of embryonal origin and lower in tumors of the glial series. The AI/mitotic index (MI) ratio was lower in malignant tumors and higher in benign tumors, suggesting a relationship between apoptosis and cell proliferation. There was no significant correlation of either AI or AI/MI ratio with either labeling index (LI) of Ki-67 clone MIB-1 or with survival. A trends towards low AI/MI ratio in tumors with high LI and short survival was observed. Apoptosis expresses cell loss in tumors, but it did not appear to be a prognostic factor. PMID- 7478274 TI - The nitric oxide synthase (NOS)-like immunoreactive extrahypophysial projections of the neurosecretory preoptic nucleus of the electric ray (elasmobranchs) suggest a neuroregulatory role for this nucleus. AB - The extrahypohysial projections of the neurosecretory preoptic nucleus (PON) of the electric ray were studied with the aid of an antibody against nitric oxide synthase (NOS). PON neurons were the only NOS-like-immunoreactive (NOS-ir) cells in the brain. These neurons gave rise to both hypophysial and extrahypophysial NOS-ir projections. Some fibres coursed from the PON to the neurointermediate lobe in the preoptic-hypophysial tract. Other NOS-ir fibres coursed either rostrally or caudally forming terminal fields in the telencephalon (subpallial region), diencephalon (preoptic nucleus, ventrolateral thalamus and posterior recess nucleus), tuberal region (area tegmentalis ventralis and substantia nigra), mesencephalon (lateral tegmentum), rhombencephalon (isthmal nucleus, vagal viscerosensory column and ventrolateral reticular area) and the spinal cord (intermediate horn). The possible involvement of the extrahypophysial PON projections in neuroregulation of visceral centres is discussed. PMID- 7478276 TI - Decompaction of CNS myelin leads to a reduction of the conduction velocity of action potentials in optic nerve. AB - The conduction velocity of action potentials in nerve fibres is proportional to degree of myelination. Here we studied the influence of myelin ultrastructure on the compound action potential conduction velocity in optic nerves of the proteolipid protein (PLP)-deficient mouse model, which displays loose myelination in central fibres. We show that a myelin decompaction leads to a suboptimal conduction velocity. The significance of myelin ultrastructure for conduction of action potential in the optic nerve is discussed. PMID- 7478277 TI - Reaction time and acoustic startle in normal human subjects. AB - We studied the effects of collision between a voluntary command and the startle response by interrupting a simple visual reaction time task with an acoustic startle. We observed two main effects. First, the reaction time was markedly shortened when the startle was delivered at intervals of 0-75 ms after the 'go' signal. Second, the startle response elicited when the subject's attention was focused onto reacting to the visual 'go' signal involved more muscles and induced larger EMG responses than when the startling stimulus was delivered unexpectedly. Startle response and voluntary commands may share a number of structures along the motor path. Reaction time can be shortened up to the latency of a startle response if the voluntary command is issued a short time after the startling stimulus. Facilitation of the startle responses during motor preparation may be due to the excitability enhancement that occurs in various neural structures along the motor path before the onset of EMG activity. PMID- 7478278 TI - Professional liability. PMID- 7478279 TI - Advance directive for health care. The Medical Society of New Jersey. PMID- 7478280 TI - How to succeed in business. PMID- 7478281 TI - Letter to the Director of New Jersey Medicaid. PMID- 7478282 TI - UMDNJ and managed care. AB - Managed care organizations and academic medical centers initially have been at opposite ends of the cost containment spectrum. UMDNJ, the largest free-standing health care university in the United States, responds to the competitive managed health care market. PMID- 7478283 TI - A survey of pediatric office-based interventions on smoking. AB - A mail survey of pediatrician practices and procedures for the modification of environmental tobacco smoke (ETS) and cigarette smoking in patients and parents was mailed to pediatricians. Results indicated that a large proportion of pediatricians address the issue of ETS and cigarette smoking. PMID- 7478284 TI - Attitudes of physicians regarding physician-assisted suicide. AB - A survey of New Jersey physicians reveals no clear consensus as to whether physician-assisted suicide should be legalized, and most physicians stated that they would not participate in such activities where it is legal to do so. Further studies need to be undertaken. PMID- 7478285 TI - The future of health care and the medical profession in the United States. AB - The medical profession is going through a traumatic period characterized by loss of autonomy and subversion of doctor-patient relationships. To prepare for the future, physicians need a shared vision and cohesive strategies and goals. Physicians need to be informed and proactive. PMID- 7478286 TI - Case report: autoimmune polyglandular syndrome. AB - The authors present a case of autoimmune polyglandular syndrome, a rare inherited condition. Early recognition allows prevention of significant morbidity and mortality. A review of the literature of this multiple endocrine end organ disease is presented. PMID- 7478287 TI - The biosynthesis of C5-C20 terpenoid compounds. PMID- 7478288 TI - Vestibular integrators in the oculomotor system. AB - The interstitial nucleus of Cajal (INC) and the nucleus prepositus hypoglossi (nph) are key elements in the vertical and horizontal oculomotor neural integrators, respectively. In this article, we attempt to develop possible circuits for these vestibular integrators by synthesizing recent information on the properties and connections of neurons involved in the integration process. We also examine how the cerebellar flocculus could play a role in the vertical integrator and vestibulo-ocular reflex (VOR) as well as in the modulation and plasticity of the VOR. We suggest that the circuitry for the vertical integrator involves the cerebellar flocculus in addition to the already proposed circuits distributed between the INC and the vestibular nuclei. The horizontal vestibular integrator is also distributed and seems to be characterized by functional compartmentalization. Both integrators play a wider role than simply transforming velocity-coded signals into position commands and may be pivotal in the short- and long-term modulation of the various oculomotor subsystems. PMID- 7478289 TI - Analysis of Ca2+ homeostasis in neurons dissociated from rat nucleus basalis. AB - Whole-cell patch-clamp recordings of calcium currents (ICa) and fura-2 microfluorimetric measurements of intracellular free Ca2+ concentration ([Ca2+]i) were made simultaneously in neurons acutely dissociated from rat nucleus basalis. Depolarization activated ICa and caused an increase in [Ca2+]i. The relationship between total Ca2+ influx and the increase in [Ca2+]i was studied. After repolarization, [Ca2+]i recovered to control values within a few seconds. A mathematical model was constructed to simulate the mechanisms underlying [Ca2+]i regulation; the parameters were (1) the rate of Ca2+ influx, (2) the rate of the [Ca2+]i increase by the Ca2+ influx, and (3) the rate of Ca2+ clearance from cytosol due to extrusion across the plasma membrane and sequestration into calcium storing organelles. After an appropriate evaluation of parameter values from the experimental results, the model mimicked the processes of [Ca2+]i increase and recovery. The experimental results and simulations suggest that (1) neurons possess a large Ca2+ buffering capacity, (2) systems for Ca2+ clearance are activated by the Ca2+ influx in a saturable manner, (3) the rate of Ca2+ clearance is relatively small compared to the rate of Ca2+ influx evoked by depolarizations, and (4) the shoulder in the [Ca2+]i recovery phase is due to the asymptote of the Ca2+ clearance rate. PMID- 7478290 TI - Distribution of neurofilament proteins and peripherin in the rat pituitary gland. AB - The distribution of neurofilament proteins and peripherin in the pituitary gland of the rat was studied with a panel of monoclonal and polyclonal antibodies recognizing different neurofilament subunits. In the posterior lobe, a dense plexus of neurofilament- and peripherin-immunoreactive fibers was seen. In the intermediate lobe neurofilament- and peripherin-immunoreactivity was seen only in nerve fibers in the connective tissue septa, while no immunoreactivity was seen in parenchymal nerve fibers. Bilateral sympathetic ganglionectomy did not affect peripherin-immunoreactivity, indicating that the peripherin-immunoreactive fibers are of central origin. In the anterior lobe, a few solitary neurofilament- and peripherin-immunoreactive fibers were observed. Western blotting confirmed the presence of 150 kD and 200 kD neurofilament proteins in the posterior lobe. No neurofilament protein was detected in the intermediate and anterior lobes. Abundant intermediate filaments were seen with electron microscopy in the nerve fibers of the connective tissue septa in the intermediate lobe. In the parenchymal nerve fibers only microtubules were seen, indicating that the lack of neurofilament immunoreactivity is due to absence of neurofilaments. PMID- 7478291 TI - ERK2-type mitogen-activated protein kinase (MAPK) and its substrates in postsynaptic density fractions from the rat brain. AB - Mitogen-activated protein kinase (MAPK) and MAPK kinase (MAPKK) were detected by Western blotting in the synaptic fraction prepared from the rat brain. There were two bands immunoreactive to the anti-MAPK antiserum in the soluble, P2, synaptosome, and synaptic plasma membrane fractions. These immunoreactive bands possibly corresponded to extracellular signal-regulated kinase (ERK) 1 and 2 (Boulton et al., 1991b), respectively. Only ERK2 was detected in the postsynaptic density (PSD) fraction. We then surveyed MAPK substrates in the synaptic fractions using purified Xenopus MAPK (ERK2-type MAPK), and found a number of MAPK substrates unique to the PSD fraction. Thus, ERK2 is present in the synapse, especially at the postsynaptic site, and it may play a role(s) in synaptic function via the phosphorylation of synapse-specific substrates. Developmental changes in ERK2 also supported its role in the synapse. PMID- 7478292 TI - Calcitonin gene-related peptide immunoreactivity in the trigeminal ganglion of Trimeresurus flavoviridis. AB - Crotaline snakes, which have infrared-sensitive pit organs, provide a good model for linking neuron morphology with sensory modality. In the trigeminal ganglion of the habu, Trimeresurus flavoviridis, cells positive for calcitonin gene related peptide-like (CGRP) immunoreactivity were found to be of two types, darkly stained and lightly stained. They were pseudo-unipolar, having an axon divided into stem, peripheral branch, and central branch, all of which were 1 micron or less in diameter. Other, CGRP-negative cells in the ganglion were also pseudo-unipolar, but much larger. In configuration, some of the positive cells were similar to the neurons with A-delta fibers, and others to the neurons with C fibers that have been reported by other workers. On the basis of their distribution and density, and physiological studies by other workers, the CGRP positive cells were judged to be not part of the infrared-receptive system, but to be involved in the transmission of nociception in small fibers. PMID- 7478293 TI - Somatocardiovascular reflexes in anesthetized rats with the central nervous system intact or acutely spinalized at the cervical level. AB - The effects of noxious mechanical stimulation of various segmental areas on heart rate and mean arterial blood pressure (MAP), as well as cardiac and renal sympathetic nerve activities were examined in anesthetized rats with the central nervous system (CNS) intact or acutely spinalized at the cervical level. In CNS intact rats, pinching for 20 s applied to any segmental skin area, but particularly that of the paw, produced an increase in heart rate, blood pressure and the sympathetic nerve activities. In acutely spinalized rats, pinching the chest, abdomen and back of the body produced large increases, while hindlimb and perineum stimulation induced only a small increase or no increase in heart rate, blood pressure and the sympathetic nerve activities. Stimulation of the right side produced particularly large responses in heart rate and stimulation of the ipsilateral side produced large responses in cardiac and renal sympathetic nerve activities in spinalized rats. These results suggest the existence of the two types of reflex responses, supraspinal and propriospinal, in the somatocardiovascular reflex. The supraspinal one has characteristics of diffuse reflex organization, while the propriospinal one has strong segmental and lateral organization. PMID- 7478294 TI - Effects of L-DOPA on fusimotor control of triceps surae muscle spindles in the cat. AB - The experiments were performed on lightly alpha-chloralose anaesthetised and spinalized cats. Alterations in fusimotor activity were assessed by recordings from single spindle afferents (90 primary and 12 secondary) from the triceps surae muscle, before and after i.v. administration of L-beta-3,4 dihydroxyphenylalanine (L-DOPA). The effects of L-DOPA on fusimotor reflexes from ipsi- and contralateral hind limb afferents were investigated by using extensions of the intact contralateral hind limb and tonic stretches of the ipsilateral posterior biceps and semitendinosus muscles as reflex stimuli. Prior to injection of L-DOPA, a low reflex responsiveness was found to both the ipsi- and the contralateral stimulation. After administration of L-DOPA, the reflex responsiveness as well as the resting activity of the muscle spindle afferents were increased as a result of enhanced activity in mainly dynamic fusimotor neurones. The results indicate that changes in fusimotor activity elicited after administration of L-DOPA are caused by release of transmission in interneuronal pathways mediating ipsi- and contralateral reflexes to mainly dynamic fusimotor neurones. The possible role of monoaminergic descending control of fusimotor neurones in the regulation of muscle tone, tremor and rigidity is discussed. PMID- 7478295 TI - Somatosensory and visual correlation in the optic tectum of a python, Python regius: a horseradish peroxidase and Golgi study. AB - In snakes with infrared receptors the optic tectum receives infrared input in addition to visual and general somatosensory inputs. In order to observe their tectal termination patterns in ball pythons, Python regius, we injected horseradish peroxidase (HRP) into the nucleus of the lateral descending trigeminal tract (LTTD) which mediates infrared information, the optic nerve, and the nucleus of the trigeminal descending tract (TTD) which relays general somatosensory information. Fibers from LTTD were found in layers 5-13 of the contralateral optic tectum, and were especially dense in layers 7a-8. Optic nerve fibers terminated in layers 7a-13 of the contralateral tectum, and mainly in layers 12-13. TTD fibers were few, and could be seen in only the rostral half of the contralateral tectum. These fibers were found in layers 5-7b, but mainly in layers 6-7a. Among various types of neurons stained by the Golgi-Cox method, we focused on six types of neurons whose dendritic arborization overlapped with the distribution of the terminals of these sensory afferents described above. It is possible that these different sensory modalities converge on a single neuron of the various types. PMID- 7478296 TI - Influences on the gamma-muscle spindle system from muscle afferents stimulated by increased intramuscular concentrations of bradykinin and 5-HT. AB - There is evidence that static muscular contractions induce a release of bradykinin (BK) in the working muscle, and that increased concentration of BK and 5-HT in a muscle increases the discharge rate of a subpopulation of group III and group IV muscular afferents. It is also known that activity in group III and IV muscle afferents may activate gamma-motoneurones to both homonymous and heteronymous muscles. The aim of the present study was to investigate whether increased concentration of BK and 5-HT in one muscle may influence the activity in primary and secondary muscle spindle afferents (MSAs) from the chemically affected muscle and from surrounding muscles, via fusimotor reflexes. The experiments were made on six cats anaesthetised with alpha-chloralose. The triceps surae (GS) and the posterior biceps and semitendinosus (PBSt) muscles were subjected to sinusoidal stretches. Simultaneous recordings of 2-11 MSAs from these muscles were made and the mean rate of firing and the modulation for each MSA were determined. Responses of 47 MSAs (26 PBSt and 21 GS) were recorded. The responsiveness of the MSAs to injections of BK (9-100 mg/ml, 0.5-1.0 ml) and 5-HT (25-150 mg/ml, 0.5-1.0 ml) was 89% and 83%, respectively, for injections into the arterial supply of the ipsilateral GS muscle, and 84% and 40% respectively for injections to the contralateral GS muscle. Of 10 secondarLy MSAs, only one was unresponsive to BK injections, while several MSAs responded to both ipsilateral and contralateral BK injections.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7478297 TI - Cerebellum of the adult reeler mutant mouse contains two Purkinje cell populations with respect to gene expression for the N-methyl-D-aspartate receptor channel. AB - Recent studies have identified five NMDA receptor subunits, which exhibit distinct cellular expressions in the normal rodent brain. The purpose of this investigation is to clarify the molecular-anatomical organization in the cerebellum of the reeler mutant mouse, in which various categories of the Purkinje cells are present as to the cell position and synaptic connectivity. In comparison with the distribution of the inositol 1,4,5-trisphosphate receptor mRNA, a molecular marker specific to the Purkinje cells, the epsilon 1 subunit mRNA of the NMDA receptor channel was found in the adjacent sections to be expressed in a subset of the Purkinje cells. In the rostrocaudal extent, the Purkinje cells expressing the epsilon 1 subunit mRNA were distributed preferentially in the rostral cerebellum, irrespective of the normal and heterotopic positions. In the mediolateral extent, they formed segregated cell clusters, interposed by epsilon 1 subunit mRNA-negative clusters. Hybridizing signals for the zeta 1 subunit mRNA were found in all the Purkinje cell population, whereas those for the epsilon 2, epsilon 3, and epsilon 4 subunit mRNAs were not detected in the cells. These findings suggest that the reeler cerebellum is topographically compartmentalized by two subpopulations of the Purkinje cells, one expressing the epsilon 1 and zeta 1 subunit mRNAs, and the other expressing the zeta 1 subunit mRNA alone. PMID- 7478298 TI - Differences in dispersion of monkey optic nerve fibers according to their terminal layers in the lateral geniculate nucleus. AB - Dispersion of retinogeniculate fibers in the monkey optic nerve was investigated in relation to their terminal layers in the lateral geniculate nucleus (LGN) using a retrograde axonal tracer (wheat germ agglutinin conjugated to horseradish peroxidase, WGA-HRP). Dispersion of fibers projecting to the parvocellular layer of the LGN (LGNp) is clearly less extensive compared with that of fibers projecting to the magnocellular layer of the LGN (LGNm) in the most distal part of the optic nerve. The occupation rate of retrogradely labeled fibers projecting from the periphery of the nasal retina to the LGNp increases only 1.2 times in the initial part of the nerve. In contrast, the occupation rate increases up to 1.6 times for labeled fibers projecting from a similar part of the nasal peripheral retina to the LGNm in the same part of the optic nerve. The morphological features of the labeled ganglion cells indicate that these fibers arise from P beta and P alpha ganglion cells, respectively. PMID- 7478299 TI - Role of basal ganglia in behavioral learning. AB - Recent studies on single neuron activity revealed that activities of many basal ganglia neurons are dependent upon the behavioral context. This may give us a reason why observed movement-related activity in the basal ganglia occurred late in relation to prime mover muscle activity. Early onset activity of primate putamen neurons was reported in a study in which animals were required to make a performance of sequential limb movements. This suggests the participation of basal ganglia in the initiation of movement in a behavioral context-dependent manner. The context-dependent activity in the basal ganglia has been shown to be acquired through learning. For instance, midbrain dopamine neurons respond to external sensory stimuli or reward only during early stages of learning motor tasks. Striate neurons acquire task-related activity through learning and the acquired activity almost disappears after selective lesions of nigrostriatal dopamine. In this article, a hypothetical scheme of basal ganglia functioning in behavioral learning is presented. Limbic input conveys information related to 'reinforcement' or 'incentive' either directly to the striatum or indirectly through nigrostriatal dopamine system, and it contributes to the acquisition and expression of learned activity in the striatum. The expression of learned striatal activity would contribute to the initiation of learned motor behavior. PMID- 7478300 TI - Correlation between anti-ubiquitin immunoreactivity and region-specific neuronal death in N-methyl-D-aspartate-treated rat hippocampal organotypic cultures. AB - Neuronal degeneration appears to be associated with changes in anti-ubiquitin immunoreactivity (UIR). To elucidate the relationship between the two events, we examined the time course of changes in UIR in pyramidal neurons of hippocampal organotypic cultures following exposure to an excitotoxin, N-methyl-D-aspartate (NMDA). In nontreated cultures, weak UIR was confined to the nucleus. Exposure to 100 microM NMDA for 15 min induced degeneration of pyramidal neurons, within 24 h, in the CA1 and CA3c regions. In these neurons, the nuclear UIR was reduced, and instead, UIR developed in the cytoplasm. In response to the same procedure, CA3a,b pyramidal neurons showed slight shrinkage but otherwise virtually normal morphological features. Little perikaryal (cytoplasmic) UIR developed in CA3a,b neurons. Both degeneration and perikaryal UIR were observed in CA3a,b neurons, however, when the culture was exposed to 300 microM NMDA. Immunoblot analysis showed that changes in the amount of a ubiquitin protein conjugate (24 kDa), presumably ubiquitinated histone, are similar to those of nuclear UIR in the same time course. We propose that the changes in the expression of nuclear and perikaryal ubiquitinated proteins represent some process closely related to neuronal death. PMID- 7478301 TI - Release of false transmitter serotonin from the dopaminergic nerve terminals of the rat pituitary intermediate lobe. AB - Rat pituitary intermediate lobe contains two types of serotonin-immunoreactive nerve terminals. Most of them are dopaminergic, in which serotonin acts as a false transmitter, while the rest are true serotoninergic nerves. In the present study, release of the false transmitter serotonin from the dopaminergic nerve terminals was studied by loading the neurons in vivo with serotonin precursor L tryptophan and MAO inhibitor pargyline, which results in accumulation of false transmitter serotonin. Subsequently pituitary neurointermediate lobe complexes were incubated in the presence of various agents. Potassium induced dramatic release of serotonin. This release was Ca(2+)-dependent, as demonstrated by an inhibition by Mg2+, and transporter-independent, since it was unaffected by GBR 12909 (a dopamine transport inhibitor). Tyramine and sodium nitroprusside, a nitric oxide donor, caused slight to remarkable release of serotonin. This release was inhibited by GBR 12909, suggesting that it was transporter-dependent. Presynaptic stimulation with apomorphine or haloperidol, dopamine receptor agonist or antagonist, respectively, or isoproterenol, agonist of the beta adrenergic receptor, did not significantly release serotonin. Thus, it seems that presynaptic receptors per se cannot induce release of significant amounts of serotonin from the IL dopaminergic fibers. Our results suggest that false transmitter serotonin in the IL dopaminergic nerve terminals is released primarily by the classical exocytotic release mechanism, but may also be partly released by the transporter-dependent, non-exocytotic release. PMID- 7478302 TI - The inhibitory role of nitric oxide (NO) in the somatocardiac sympathetic C reflex in anesthetized rats. AB - The role of nitric oxide (NO) in the two somatosympathetic reflex arcs, i.e. A- and C-reflexes, was examined using NO synthase (NOS) inhibitor in anesthetized rats. The A- and C-reflex components were recorded from a cardiac sympathetic efferent nerve and elicited by stimulation of myelinated A and unmyelinated C afferent fibers in the left tibial nerve. NG-nitro-L-arginine methyl ester (L NAME), a NOS inhibitor, when administered by either intrathecal (i.t.) or into the cisterna magna (i.c.m.) routes, augmented only the C-reflex in a dose dependent manner. The effective i.t. dose of L-NAME to augment the C-reflex was approximately 1000 times the i.c.m. dose. NG-nitro-D-arginine methyl ester (D NAME), an isomer of L-NAME, had no effect on either A- or C-reflexes, when administered i.c.m. Neither i.c.m. pre-treatment nor post-treatment with L arginine, a NOS substrate, influenced either A- or C-reflexes, but i.c.m. pre treatment with L-arginine abolished the facilitatory effect of L-NAME on the C reflex. These results suggest that NO, synthesized in the brain stem, plays an inhibitory role in the central modulation of the somatocardiac sympathetic C reflex. The possibility of movement of L-NAME to the brain stem from the spinal cord is discussed. PMID- 7478303 TI - An anterograde-retrograde labeling study of the carotid sinus nerve of the Japanese monkey (Macaca fuscata). AB - The sites of origin and termination of efferent and afferent fibers in the carotid sinus nerve (CSN) were investigated in the Japanese monkey. After application of a mixture of horseradish peroxidase (HRP) and wheat germ aggulutinin-conjugated HRP to the central cut end of the CSN, sensory ganglion neurons were labeled in the jugular ganglion of the vagus nerve, as well as in the superior and petrosal ganglia of the glossopharyngeal nerve. Many sympathetic ganglion neurons were also labeled retrogradely in the superior cervical ganglion. In the brain, many labeled terminals were seen ipsilaterally in the lateral division of the nucleus of the solitary tract (NST). A few neuronal cell bodies were also labeled ipsilaterally in a reticular region dorsomedial to the caudal one-third of the facial nucleus. The results indicate that the CSN of the Japanese monkey is composed mainly of afferent fibers terminating in the NST, that the afferent fibers in the CSN originate not only from the superior and petrosal ganglia of the glossopharyngeal nerve but also from the jugular ganglion of the vagus nerve, and that efferent fibers contained in the CSN arise from the medullary reticular formation and superior cervical ganglion. PMID- 7478304 TI - Three-dimensional anisotropy contrast magnetic resonance imaging of the rat nervous system: MR axonography. AB - Anisotropy of apparent diffusivities (Dapp) of water molecules is known to be an excellent means for elucidating detailed information regarding neuronal fiber direction and density in live subjects (axonography). Nevertheless, determination of eigenvalues and eigenvectors of the apparent diffusion tensor, D xi app, the necessary process for the treatment of D xi app in axonography, is cumbersome and the quality of the reconstructed images based on estimated parameter values of D xi app has often been disappointing. We present here a simple algorithm which allows for the extraction of the anisotropic term vector, DAapp, from D xi app in pictorial form, which in turn provides a novel color-coded contrast for magnetic resonance imaging (MRI). MRI images constructed using this technique, three dimensional anisotropy contrast (3DAC), provided impressive anatomic resolution of the CNS, comparable to in vitro histology. Directional information of DAapp expressed by hue of color clearly correlated to fiber direction, while magnitude of DAapp expressed by intensity of color correlated to fiber diameter. Magnetic resonance axonography (MRX) utilizing 3DAC technique possesses significant potential for various biological and clinical applications. PMID- 7478305 TI - Participation of cAMP and cAMP-dependent protein kinase in beta-adrenoceptor mediated interleukin-1 beta mRNA induction in cultured microglia. AB - We previously reported evidence of beta-adrenoceptor-mediated induction of IL-1 beta mRNA in the rat hypothalamus. The present in vitro studies using northern blot analysis showed that the beta-adrenoceptor agonist isoproterenol (1 x 10(-8) to 1 x 10(-5) M) caused a marked induction of IL-1 beta mRNA in microglia, but not in astrocytes. This induction was remarkably suppressed by pretreatment of cells with the beta-adrenoceptor antagonist propranolol. These phenomena were confirmed by in situ hybridization with digoxigenin-labelled IL-1 beta RNA probe. Furthermore, dibutyryl cyclicAMP (dbcAMP) (5 x 10(-4) and 5 x 10(-5) M) markedly induced IL-1 beta mRNA in microglia. The intracellular level of cAMP in microglia was elevated in a dose-dependent manner when they were treated with isoproterenol, and this elevation was completely blocked by propranolol. The induction of IL-1 beta mRNA by either isoproterenol or dbcAMP was strongly inhibited by a cAMP-dependent protein kinase inhibitor, H8. These results, taken together, suggest that (1) microglia primarily induce IL-1 beta mRNA by stimulation of beta-adrenoceptors, and (2) cAMP and cAMP-dependent protein kinase presumably participate in a signal transduction mechanism involved in the induction of IL-1 beta mRNA via beta-adrenoceptors. PMID- 7478306 TI - Neuronal expression of a minor monosialosyl ganglioside GM1b in rat brain: immunochemical characterization using a specific monoclonal antibody. AB - Monoclonal antibodies (mAbs) against GM1b ganglioside were raised by immunizing NZB/n mice with the antigen purified from bovine brains, and the details of binding specificity of the mAbs were characterized. Anti-GM1b mAb, termed NA-6, reacted specifically with GM1b (NeuAc) and GM1b(NeuGc). NA-6 antibody did not react with other structurally related gangliosides, indicating that the antibody recognizes NeuAc or NeuGc alpha 2-3Gal beta 1-4GalNAc beta 1-4Gal structure. Using NA-6 antibody, GM1b ganglioside in developing rat brain was investigated by TLC/enzyme-immunostaining and detected first on gestational day 16. The specific content of brain GM1b increased until postnatal day 10, and then gradually decreased in later stage of development. Immunohistochemically GM1b was found in proximal dendrites and cell bodies of neurons in extensive regions of adult rat brain. The immunoreactivity tended to be confined in patch-like clusters on cell membranes, as typically found in the hippocampus and neocortex. The GM1b synthase activity, when assayed in the cerebellar microsome proteins, was significantly reduced in lurcher mutant mouse that is devoid of both cerebellar granule and Purkinje cells. These findings demonstrate that GM1b ganglioside exists in neurons and is actively synthesized during the development in rat brain. PMID- 7478307 TI - Rotorod sensorimotor learning in cerebellar mutant mice. AB - Lurcher mutant mice, characterized by degeneration of cerebellar granule and Purkinje cells, were compared to normal littermate controls in a rotorod test, consisting of a wheel turning at constant speed which required on the part of the animal postural adjustments in order to maintain equilibrium. Identical baseline rates for the two groups were assured by changing the speed and size of the rotating rod. Although both groups were able to learn the task, the fall latencies of normal mice exceeded those of lurchers. These results indicate that cerebellar cortical atrophy does not abolish this form of sensorimotor learning. However, brain-damaged animals are unable to reach the same level of performance as normal animals. In contrast to the results in lurcher mutants, no sensorimotor learning was displayed by hot-foot mutants and staggerer mutants. PMID- 7478308 TI - The effects of MK801 on place conditioning. AB - The appetitive properties of MK801 were investigated using the conditioned place preference paradigm. Male Sprague-Dawley rats received conditioned place preference training for a four-day period. The conditioning box consisted of three chambers with distinctive visual and tactile cues, separated by removable doors. On alternating days rats received MK801 (0.05, 0.1, 0.25, 0.5 and 0.75 mg/kg) paired with one side of the chamber and saline paired with the other side. On the fifth day, rats were placed in the center chamber and the time spent in each chamber, as well as entries into each chamber were recorded by a trained observer. MK801 produced a conditioned place preference for side paired with drug for the four highest doses. Contrary to previous findings, these results indicate that MK801 is appetitive at doses higher than has previously been assumed. PMID- 7478309 TI - Portion sizes in dietary assessment: issues and policy implications. AB - Accurate estimates of energy and nutrient intake of individuals and populations are contingent on the reliability of information obtained about food intake, food composition, and portion size. This article reviews issues related to the definition, determination, and use of portion sizes in dietary assessment and public education. PMID- 7478310 TI - Selenium in human lactation. AB - The primary factor determining selenium concentration in human milk is the maternal selenium intake. A significant correlation between selenium in human milk and maternal selenium intake has been reviewed in papers from different regions of the world. Infants fed human milk have higher selenium intake than those fed commercially available formula milk or baby foods. Selenium compounds found in breast milk seem to be more biologically available for infant nutrition than those in formulas. Increased requirements of selenium have been observed in pregnant and lactating women. Supplementation of lactating and pregnant women with different selenium compounds has been assayed, and selenium supplementation of soil and cows has been used to increase the selenium status of children fed infant formula made from cow's milk. PMID- 7478311 TI - Plasma transferrin receptor helps to predict iron deficiency in the anemia of chronic disease. AB - A recent study by Ahluwalia and colleagues used a discriminant statistical analysis approach to determine that a combination of serum ferritin, plasma transferrin receptor concentration, and erythrocyte sedimentation rate was the optimal set of variables for differentiating iron deficiency and the anemia associated with chronic disease in a group of elderly women. Iron deficiency was defined as a significant response in hemoglobin concentration after iron supplementation. The findings of this study suggest that iron deficiency can be relatively common among elderly anemic women with rheumatoid arthritis. Use of these three biochemical measures should be clinically useful to differentiate iron deficiency in the anemia of chronic disease. PMID- 7478312 TI - The iron-copper connection: the link to ceruloplasmin grows stronger. AB - A rare genetic defect in ceruloplasmin biosynthesis has provided the strongest evidence to date that ceruloplasmin is essential for iron metabolism and tissue distribution in humans. A aceruloplasminemia results in massive accumulations of iron in the liver and brain and is associated with retinal degeneration and diabetes. PMID- 7478313 TI - Is an elevated circulating maternal homocysteine concentration a risk factor for neural tube defects? AB - The mechanism by which folic acid supplementation reduces neural tube defect (NTD) incidence is unknown. Recent evidence suggests that mothers with NTD neonates have higher circulating homocysteine concentrations compared with controls. This presumably indicates impaired homocysteine remethylation, resulting in a possible methionine shortage at a crucial stage of fetal development. PMID- 7478314 TI - How to chart home health nursing. PMID- 7478315 TI - How to foil the flu. PMID- 7478316 TI - Self-test. Understanding antibacterials and antivirals. PMID- 7478317 TI - Transferring a patient from bed to chair. PMID- 7478318 TI - Treating urinary incontinence with timed schedules and exercises. PMID- 7478319 TI - Patient-teaching aid. Taking your medication correctly. PMID- 7478320 TI - Delegating successfully. PMID- 7478321 TI - Radiofrequency catheter ablation. Controlling tachyarrhythmias. PMID- 7478322 TI - Actionstat. Repeated ICD discharges. PMID- 7478323 TI - Cerebral edema: bringing the brain back down to size. PMID- 7478324 TI - After a brain injury: clearing up the confusion. PMID- 7478325 TI - A second chance for Kirk. PMID- 7478326 TI - Critical thinking. Taking the road less traveled. PMID- 7478327 TI - Refilling an implantable pump. PMID- 7478328 TI - What Mrs. Jerome smelled. PMID- 7478329 TI - Assessing flank pain. PMID- 7478330 TI - Remembering Rachael. PMID- 7478331 TI - The prevention of relapse in bipolar disorder. PMID- 7478332 TI - Lead in children from older housing areas in the Wellington region. AB - AIMS: To examine blood lead levels in children, aged 12 to 23 months, living in old housing areas of Wellington and Lower Hutt, and to investigate risk factors for high lead levels. METHOD: Children were selected from Plunket Society rolls. Venous blood samples were collected, and care givers were interviewed with a questionnaire. Soil samples were taken from around the children's homes. Both soil and blood samples were analysed for lead content. RESULTS: Blood samples and completed questionnaires were obtained for 143 children. The geometric mean blood lead level for all the children was 0.25 mumol/L (5.1 micrograms/dL) (95% confidence interval [95% CI]: 0.22-0.28 mumol/L). Three children had blood lead levels that exceeded the level for notification in New Zealand 1.45 mumol/L and a further 13 had blood lead levels exceeding 0.48 mumol/L. Children with elevated lead levels were likely to live in a house greater than 50 years old where paint removal had taken place in the last 2 years (risk ratio [RR] = 14.4, 95% CI: 2 107). Eating dirt, particularly for children who usually played outside within 2 metres of the house, was also a risk factor for elevated blood lead levels. Soil lead levels generally increased with the age of the house and were weakly correlated with blood lead levels (r = 0.32). CONCLUSION: Paint removal in old houses is a major risk factor for elevated blood lead levels. However, the number of study children living in houses less than 50 years old was limited. Because of this and possible participant selection bias, the results of this study require confirmation in a separate population-based study. Information about the specific paint removal procedures that cause high lead levels is also needed. PMID- 7478333 TI - Fragile X mental retardation syndrome: DNA diagnosis and carrier detection in New Zealand families. AB - AIMS: To establish a DNA-based test for the diagnosis and carrier detection of fragile X syndrome, and to investigate the nature of the mutation and patterns of inheritance in New Zealand families. METHODS: A probe for the FRAXA region was generated by polymerase chain reaction, cloned in a plasmid vector, and its structure was confirmed by DNA sequencing. This probe was used in a Southern blot assay to detect full mutations or premutations associated with fragile X syndrome in DNA from peripheral blood samples submitted to our laboratory for routine testing. RESULTS: We tested 379 individuals from throughout New Zealand. Full mutations were found in 29 males, leading to a fragile X diagnosis, or confirmation of an earlier cytogenetic diagnosis. Premutations were detected in 45 females and 11 males, all of whom are asymptomatic carriers of the disease. CONCLUSIONS: The DNA test is rapid and accurate, in contrast to the cytogenetic test. It allows unequivocal detection of carriers, enabling effective counselling, prenatal testing, and more generalised screening of at-risk populations. Our discovery of one large pedigree with many carriers and no prior history of X-linked mental retardation demonstrates that the DNA test is appropriate even in apparently sporadic cases of mental retardation. PMID- 7478334 TI - Where are tomorrow's specialists? AB - AIM: To determine the extent to which New Zealand's future specialists are likely to be graduates of this country's medical schools. METHODS: Forward projections were made on the basis of information from: (1) Medical Council annual reports recording additions to the Register of Specialists between July 1 1990 and June 30 1994, (2) Medical Council annual questionnaire data relating to New Zealand doctors qualifying in 1971-85, (3) Medical Directories of Australia, Canada, UK and USA, (4) selected Australasian specialist colleges on the number of registered trainees in New Zealand, and (5) Medical School data on New Zealand graduating classes since 1971. RESULTS: Forty percent of the new specialists registered in New Zealand between 1 July 1990 and 30 June 1994 were graduates of overseas medical schools. Future projections suggest that this percentage is likely to increase. Major determinants of New Zealand specialist numbers are (1) the proportion of specialist New Zealand graduates (38%) not working in this country, and (2) the sex ratio of future graduating classes--proportionately fewer women (38%) than men (56%) specialise. CONCLUSION: Graduates of this country's medical schools are likely to supply approximately half of New Zealand's future specialists. PMID- 7478336 TI - An acute compartment syndrome involving the anterior thighs after isometric exercise. PMID- 7478335 TI - The results of two years operation of the visiting specialist service to the Waikato mobile ear clinic. AB - AIM: To review the function and results of the Health Waikato mobile ear clinic service 1993-4. METHOD: Data were collected on all children reviewed at the fortnightly specialist clinics conducted in the mobile ear clinics during 1993-4. An overall pathological assessment of the middle ear and eardrum was made for each ear. RESULTS: Of the 467 children examined 311 children were wait listed for elective surgery. Pathology ranged from recurrent acute otitis media to cholesteatoma. Large numbers of children were found with atelectasis of the tympanic membrane. Children as young as seven years of age required modified radical mastoidectomy for treatment of otorrhea due to cholesteatoma. CONCLUSIONS: The results of this survey show the mobile ear clinic service requires a large amount of resources to operate. The mobile ear clinic generates a large operative workload which creates an imbalance between resources devoted to finding the disease and those devoted to treating it. Recommendations include a tighter integration between the mobile ear clinic and the hospital, and appropriate allocation of treatment services to treat the children identified by the service. PMID- 7478337 TI - Laparoscopic inguinal hernioplasty. PMID- 7478338 TI - Carriage of N meningitidis. PMID- 7478340 TI - Enquiring about test results. PMID- 7478339 TI - A direct threat to research in public hospitals. PMID- 7478341 TI - Stroke rehabilitation. PMID- 7478342 TI - Stroke units. PMID- 7478343 TI - Smokers run enormous risk: new evidence. PMID- 7478344 TI - Hysterectomy for nonmalignant conditions: cost to New Zealand society. AB - AIMS: The aim of the study was to estimate the annual hospital costs to New Zealand society of hysterectomy for nonmalignant conditions in women between puberty and menopause, and to compare international rates for all hysterectomies. METHODS: The records of patients aged between 15 and 54 years inclusive, who had a hysterectomy for a nonmalignant menstrual condition, were researched. Hospital costs were estimated using specific hospital unit costs and prices. RESULTS: The 4390 hysterectomies for nonmalignant conditions represent 69% of the total hysterectomies undertaken in New Zealand in 1991. Estimated costs for all hospitals were $17 million or $3868 per hysterectomy. The all ages all conditions hysterectomy rate per 100,000 women of 365 (368 in 1992) in New Zealand is higher than in some other developed countries. CONCLUSIONS: Further reduction in the rate of hysterectomy in New Zealand may save hospital costs but these savings should be offset against the costs of any other treatment options chosen. Although it was found that unit costs of public hospitals were higher than those of private hospitals the case mix for the two types of hospital may differ and public hospital cost reductions may not be possible. Economic evaluation of alternative treatment options would be improved if all relevant costs (direct, indirect and intangible) were known. Further research is needed to determine the reasons why New Zealand women elect to have hysterectomies, why most choose a private hospital, and changes (before and after hysterectomy) in productivity, out of pocket expenses, and quality of life for these patients. PMID- 7478345 TI - Hip fracture incidence among older people in Auckland: a population-based study. AB - AIM: To determine the incidence of hip fractures among those aged 60 years or older in the Auckland region. METHODS: Demographic data were obtained on all older people hospitalised with a fracture of the femoral neck in Auckland, between 8 July 1991 and 7 February 1994. Incidence rates were calculated using the 1991 census data. RESULTS: An average incidence of 733 hip fractures or 710 individuals sustaining hip fractures each year was observed. Age-adjusted incidence rates were 571.5 per 100,000 for women and 318.2 per 100,000 for men. 97.0% of the hip fractures were sustained by those identified in the records as European, with the rates for Maori and Pacific Island populations 25-50% lower than for Europeans. Among Maori and Pacific Island populations, incidence rates were similar for men and women. Overall, 41.8% of the hip fractures were sustained by those > or = 85 years, with 67.0% sustained by those > or = 80 years. Incidence rates increased with age, with the rates continuing to increase in those > or = 85 years. CONCLUSION: Among Europeans, hip fracture incidence rates in Auckland are similar to those found for other caucasian populations. However, the rates among Maori and Pacific Island populations are among the lowest internationally and are similar for men and women. The increase in incidence rates with age continues in those > or = 85 years. PMID- 7478347 TI - Court ordered obstetric intervention: a commentary. AB - A case is presented where the Courts have authorised an obstetric intervention deemed necessary for the well-being of both mother and child. Although the case is one of maternal psychosis, there are legal and ethical concerns whenever court ordered intervention is deemed necessary. Approaches to this difficult medical decision making problem in the form of utilitarian "burdens v benefit" ratio analysis or the recognised traditional ethical principles of beneficence, nonmaleficence, justice and acting in the patient's best interest are considered. The Royal College of Obstetricians and Gynaecologists guidelines suggesting "that it is inappropriate ... to invoke judicial intervention to overrule an informed and competent woman's refusal of a proposed medical treatment, even though her refusal might place her life and that of her fetus at risk" are questioned. PMID- 7478346 TI - Alcohol and pregnancy: doctors' attitudes, knowledge and clinical practice. AB - AIMS: This study was undertaken to gain an understanding about doctors' perspectives on alcohol consumption in pregnancy. The specific aim was to examine attitudes, knowledge and clinical practice of a sample of New Zealand doctors. METHODS: A questionnaire was sent to all paediatricians, all obstetricians and a random sample of general practitioners throughout New Zealand. RESULTS: All respondents felt there should be a limit on alcohol consumption in pregnancy and 46% recommended abstinence. Doctors perceived public awareness of the risks of alcohol consumption in pregnancy to be poor and over 90% believed awareness needed to improve. Eighty-nine per cent of obstetricians and 84% of general practitioners reported they felt sufficiently knowledgeable to inform people of the risks of alcohol consumption in pregnancy. Fifty-nine per cent of obstetricians and 40% of general practitioners routinely enquire about alcohol consumption at the first antenatal contact (p < 0.003). Fifty three per cent of obstetricians and 48% of general practitioners routinely advise women about alcohol consumption at the first antenatal contact. CONCLUSIONS: Doctors in New Zealand believe alcohol in pregnancy is an important health issue. Awareness of the risks of alcohol consumption in pregnancy is perceived as poor. Doctors' attitudes to alcohol consumption in pregnancy do not reflect their reported clinical practice. PMID- 7478348 TI - Herpes zoster during pregnancy: a case report. PMID- 7478349 TI - Core components for a diabetic service. New Zealand Society for the Study of Diabetes. PMID- 7478350 TI - Laparoscopic inguinal hernia repair: what your patient needs to know. PMID- 7478351 TI - Meningococcal disease epidemiology and control in New Zealand. AB - Epidemiology, surveillance and research New Zealand has a high quality surveillance system for meningococcal disease that successfully integrates notification and laboratory data. Since 1991, New Zealand has had elevated incidence rates of meningococcal disease rising to 6.2 per 100,000 population in 1994. This represents a rate that is four times that recorded in 1989/90. Serogroup B infection predominates and international experience suggests that these elevated rates may continue for 5 to 15 years. Rates of meningococcal disease in Maori and Pacific Islands populations were three times higher than in Europeans at 10.0 and 12.3 per 100,000 respectively in 1994. The rates were particularly high for infants with the rate in Maori infants under 1 year reaching 120 per 100,000. The case fatality rate at 5.3% for 1994 would appear to be relatively low by international standards. Case control studies could be used to investigate potentially modifiable primary risk factors for disease. Intensive case review studies to investigate the role of such factors as preadmission antibiotics in reducing severe outcomes may be of benefit. The Ministry of Health or research funding organisations should consider the potential value of such studies in more detail. PMID- 7478352 TI - Streptokinase audit at Thames Hospital. PMID- 7478353 TI - The prevalence of asthma. PMID- 7478354 TI - Cot death and cot mattresses. PMID- 7478355 TI - Management of patients with dyslipidaemia. PMID- 7478356 TI - Medicare cap rates up but still flawed. PMID- 7478357 TI - Merger mania, alliances and subacute care. PMID- 7478358 TI - Ethics in the information age. PMID- 7478359 TI - Home care liability--Part I. PMID- 7478360 TI - Care management: providing a connecting link. AB - Care management monitors the variances of the entire patient population in a given area versus specific cases as demonstrated in case management. It includes CQI principles, variance analysis and regulatory standards. Care management serves as a conduit for continuing education, involves a multidisciplinary team approach, which involves the family and assists managed care and utilization review. PMID- 7478361 TI - A CNS integrated health care delivery system model. AB - The more complicated a client's health needs are, the higher the risk for inappropriate utilization of services, fragmentation of care and delays in system access. The challenge is to ensure that patients move appropriately from one access point to another in the most efficient, effective manner. The Integrated Health Care Delivery System is designed to meet this challenge. PMID- 7478362 TI - Providing continuity in a "firm" case management system. AB - Four general medicine units were divided into two "firms" to improve patient outcomes and ensure continuity of care. All members remain part of their firm for as long as they are employed. The nurse case managers remain constant in both inpatient and outpatient settings and coordinate discharge planning. PMID- 7478363 TI - Comparing work satisfaction in two case management models. AB - Case management by professional nurses is expanding rapidly in managed care systems, but little is known about the quality of working life of nurse case managers (NCMs). This study explores the relationships among control over nursing practice, job stress and work satisfaction among NCMs and compares NCMs in a "within-the-walls" model with those in a "beyond-the-walls" model of nursing case management. Results of this small descriptive study suggest that NCMs perceived a relatively high degree of control over their practice and work satisfaction as well as a relatively low degree of job stress. PMID- 7478364 TI - A centralized communication system. AB - Patient care redesign involves people, processes and technology coming together to meet or exceed patient expectations. A centralized communication system promotes professional nursing practice and optimizes all the team members' roles through more efficient use of staff. PMID- 7478365 TI - A perinatal cross-training program. AB - A comprehensive program involving the orientation of perinatal staff to a secondary clinical area has led to a flexible staffing plan. After one year, the perinatal department has improved financially, while staff have improved their marketability. PMID- 7478366 TI - Director of nursing finance: controlling health care costs. AB - Vice Presidents of Nursing in 250 hospitals throughout the United States were asked whether they have someone who is responsible for the nursing division budget as well as coordinating all the department's financial matters. Job duties believed to be relevant to a Director of Nursing Finance position were explored as well as who within the hospital's organizational structure typically performs these job duties. PMID- 7478368 TI - Medicare: what's happening? PMID- 7478367 TI - Going beyond enlightened self-interest. PMID- 7478369 TI - Critical pathways in the emergency department. PMID- 7478371 TI - Orientation to the charge nurse role. PMID- 7478370 TI - Establishing a unit-based cost containment committee. PMID- 7478372 TI - Case management in a small rural hospital. PMID- 7478373 TI - A patient classification system for the hemodialysis setting. PMID- 7478374 TI - When the going gets tough, the tough... PMID- 7478375 TI - Independent contractors beware. IRS declares war on New York dentists. PMID- 7478376 TI - Resisting the routine. PMID- 7478377 TI - Managed care myth. PMID- 7478378 TI - DSSNY comes of political age. PMID- 7478379 TI - The dental clinical workstation and the computer-based patient record. AB - Computer-based clinical workstations and software systems for patients records offer emerging technologies important to the practice of dentistry. Many considerations enter into the design of the workstation: a screen graphics software design that will permit fast, accurate and comprehensive data input while being easy to learn and use; space-saving computer hardware, including the monitor, and an integrated physical design for the complete clinical workstation that takes into account infection control and ergonomics. Other computer-based technologies, such as intra-oral imaging systems and digitized radiographs, along with accounting, billing, insurance claims and scheduling software systems should be integrated with the workstation software for clinical data acquisition. This will require adoption of uniform standards for the data dictionaries and for communication with outside databases, hospitals, laboratories and service bureaus, which suggests that the clinical workstation may become the integrating centerpiece for a variety of clinical computer-based technologies that are emerging in the health professions. PMID- 7478380 TI - Computers are here. Now what do we do with them? PMID- 7478381 TI - Choosing practice management software. How sophisticated should you get? AB - In order to make informed purchasing decisions about practice management software, dental professionals must first assess their needs by determining the types of tasks they expect from a system and then compare that list against the features offered by each system. Because technology is changing rapidly, much of what was advanced a few years ago is now considered basic. Three general levels of sophistication are discussed, with attention to the complexity of features that can be expected to be found at each level. PMID- 7478382 TI - Sexual harassment in the dental office. A model policy. PMID- 7478383 TI - The dental information superhighway. Opportunity and challenge. AB - The "familiar" practice of dentistry is becoming history. The changes taking place in dentistry parallel what is happening across the world, with computer based technologies taking on prominent roles. The information superhighway is in full evidence in dentistry, and is enabling doctors, through technology, to gain instantaneous access to information and ideas. It is also altering the manner in which business is done. PMID- 7478384 TI - Multimedia approach to dental education in the 21st century. AB - Today's technology allows educators to design and create tools that enable teachers and students to work with material from different sources and to attend courses at various sites. The personal computer represents perhaps the greatest boon to this movement by providing a vehicle for "user friendly," interactive, multimedia courseware. A discussion of that courseware, its development and implementation, is provided here. PMID- 7478385 TI - Growth of the Internet and its impact on health care and society. AB - The discovery of new tools adds to the human experience, but our newfound ability to communicate and share knowledge and wisdom almost instantaneously results in a factorial increase in the collective wisdom of a global community. Computer networks continue to expand and reach into every corner of the globe. This global information network has the potential to affect many aspects of society. This paper presents a brief history and overview of the Internet and network application tools and discusses some of the ethical issues that need to be addressed as the Internet grows to touch every part of our lives. PMID- 7478386 TI - The use of mucosal inserts. AB - Mucosal inserts increase the retention of any well-made denture but should not be attempted as a substitute for good prosthetic technique. They are especially useful with the trauma or cancer patient where functional anatomy has been altered. Final results of the patient's removable prosthesis will depend on mechanical, anatomical and psychological factors. The mucosal insert denture can satisfy the expectations of many of these patients, if the patients are informed and managed properly. PMID- 7478387 TI - Tobacco takes toll on ballplayers. PMID- 7478388 TI - Specialty training in nuclear medicine in the UK. PMID- 7478389 TI - Parathyroid imaging. AB - Developments in technology mean that it is now possible to localize abnormal parathyroid glands by a variety of imaging methods. There remain a number of unresolved issues. With all commonly used methods (radionuclide imaging, ultrasound, computed tomography and magnetic resonance imaging), there are wide variations in sensitivity of detection of glands, and in many cases this variation is not explained satisfactorily. The necessity for imaging is questioned by many parathyroid surgeons, particularly before a first operation. However, parathyroid surgery is itself changing, with a trend towards more limited neck exploration, which requires localization of the glands pre operatively. This review describes the methods available for localizing parathyroid glands, with their advantages and limitations, and discusses the role of the techniques prior to surgery. PMID- 7478390 TI - Specifying dual-detector gamma cameras and associated computer systems. AB - Increasingly, dual-detector gamma cameras are being purchased as replacements for single-headed cameras. The improvement in sensitivity offered by a dual-detector device can be used in several ways: to shorten acquisition times, to improve signal-to-noise ratios, or to reduce administered doses. This paper focuses on the practical aspects of dual-headed devices and aims to provide some guidance for potential purchasers in the drawing up of an equipment specification. PMID- 7478391 TI - Objective measurement of changes in regional wall motion characteristics following coronary artery bypass grafting using tomographic radionuclide ventriculography. AB - Difficulties arise when using non-invasive methods to measure changes in regional left ventricular function. With the increasing recognition of the entity of hibernating myocardium, and the known asymmetric effects of coronary artery disease, this is an important problem, as it prevents detailed investigation of the effects of revascularization upon ventricular function. We investigated the use of tomographic radionuclide ventriculography in assessing such changes. Twenty consecutive patients (18 males, 2 females, mean age 60 years), undergoing elective coronary artery bypass surgery, were identified and imaged prior to and after surgery. The mean global left ventricular ejection fraction was 42 and 45% pre- and post-surgery, respectively. Following revascularization, it was improved in 9 patients, unchanged in 3 and deteriorated in 8. By comparison with pooled normal data from 25 subjects, 10 patients were noted to have overall resting phase values within normal limits. After surgery, an improvement was seen in 5 patients, no significant change in 11 and a deterioration in 4. With respect to regional ejection fraction, 24 of 80 segments improved, 25 remained unchanged and 31 deteriorated. For regional phase analysis, 26 improved, 45 remained unchanged and 9 deteriorated. confirmed that important regional changes in left ventricular function occur following revascularization, even without a change in global ejection fraction. PMID- 7478392 TI - Relation between dobutamine trans-thoracic echocardiography, 99Tcm-MIBI and 18FDG uptake in chronic coronary artery disease. AB - The relationships between rest conditions of myocardial asynergy, response to dobutamine administration, perfusion and glucose metabolism were examined in 12 patients with chronic coronary artery disease and left ventricular dysfunction. We evaluated (1) rest and stress myocardial perfusion by 99Tcm methoxyisobutylisonitrile (MIBI) and single photon emission tomography (SPET), (2) rest myocardial segmental wall motion by trans-thoracic echocardiography and low-dose dobutamine, and (3) myocardial metabolism by [18F]-2-fluoro-2-deoxy-D glucose (18-FDG) and positron emission tomography (PET), in the fasting state. The analysis was carried out on 16 left ventricular myocardial segments. The SPET studies were analysed semi-quantitatively by normalization to the peak activity. Wall motion was assessed by a visual score. An 18FDG index was determined as the tissue/blood pool radioactivity ratio in each segment. The results showed: (1) remarkably good agreement between the number of dobutamine responsive segments and 18FDG positive segments among those that were only moderately hypoperfused and hypokinetic; (2) a smaller number of dobutamine responsive segments than 18FDG positive segments among those that were hypoperfused and akinetic; and (3) the presence of 18FDG in 50% of the segments that were severely hypoperfused and akinetic or dyskinetic and without improvement with dobutamine. These results indicate that in severely hypoperfused and akinetic or dyskinetic segments, trans thoracic echocardiography under inotropic stimulation provides little additional information compared with that obtained with rest echocardiography and perfusion studies; the assessment of 18FDG uptake provides information that is complementary to that obtained by perfusion assessment, rest and dobutamine trans thoracic echocardiography. PMID- 7478393 TI - 99Tcm-pyrophosphate myocardial scintigraphy: the role of volume-rendered three dimensional imaging in the diagnosis of acute myocardial infarction. AB - The aim of this study was to evaluate the role of volume-rendered three dimensional imaging in the diagnosis of acute myocardial infarction (AMI) using 99Tcm-pyrophosphate (99Tcm-PYP) scintigraphy in a diagnostically difficult group of patients. In total, 117 patients were studied using planar, single photon emission tomography (SPET) and 3-D volume-rendered imaging performed 3 h after receiving 555 MBq (15 mCi) of 99Tcm-PYP intravenously. 555MBq (15 mCi) of 99Tcm PYP intravenously. Two teams of physicians reported in a blinded and random fashion on each planar, SPET and 3-D rotating image study. Individual reports were compared with reports that used all three imaging modalities in combination. Observer reproducibility was between 85 and 90% and inter-observer team agreement was between 87 and 91%. A score based on clinical history, electrocardiography, serum cardiac enzyme levels and cardiac risk factors was validated and used as a de facto 'gold standard' for AMI for 75 of the subjects for whom all these details were available. The sensitivity, specificity and overall accuracy of combined planar, SPET and 3-D rotating image modalities were all 84%. Analysis of each modality in isolation showed SPET imaging to have the highest sensitivity (74%) and specificity (89%). In combination with SPET and planar imaging, 3-D rotation image presentation increases diagnostic sensitivity without appreciably altering overall diagnostic accuracy. 99Tcm-PYP myocardial SPET imaging shows good utility in the diagnosis of AMI in diagnostically difficult patients. PMID- 7478394 TI - Absolute 99Tcm-DMSA renal uptake in children: a study of 321 kidneys. AB - Absolute 99Tcm-dimercaptosuccinic acid (DMSA) uptake was measured in 160 children ranging in age from 3 months to 15 years. In total, 108 pairs of kidneys were normal and the mean uptake for the left and right kidneys was 24.0 and 23.4%, respectively. The mean summed uptake for the left and right kidneys was 47.3%. A method of calculation based on the geometric mean of the anterior and posterior views was used to allow for radiation attenuation. DMSA uptake did not vary significantly with the age of the child. The value of the method in the assessment of abnormal kidneys is presented. The effect of using a supine or prone position for the anterior view was also investigated. PMID- 7478395 TI - Absolute 99Tcm-DMSA renal uptake in children: optimum time to scan. AB - Absolute renal uptake was measured at 2, 4 and 6 h in 27 patients in order to determine a more convenient time for uptake compared with the 6-h measurement proposed by other authors. Measurements made at 2 and 4 h would need to be increased by 20 and 6%, respectively, to convert to the value at 6 h. Measurements at 4 h are a reasonable compromise to achieve a high-count/low background image in a reasonable time and to obtain a good estimate of the predicted 6 h uptake at a single scanning session. The percent renal uptake at 6 h, U(6), can be derived from the percent uptake measured at t hours after injection, U(t), using the following formula: U(6) = U(t) x CF(t), where CF(t) = 1.0 + 0.03 (6 - t) for 4 < or = t < or = 6. PMID- 7478396 TI - Superimposition of PET images using 18F-fluorodeoxyglucose with magnetic resonance images in patients with pancreatic carcinoma. AB - Six patients with histologically proven adenocarcinoma of the pancreas were studied using 2-deoxy-2-[18F]-fluoro-D-glucose (18F-FDG) using positron emission tomography (PET), and magnetic resonance imaging (MRI). In all six cases there was avid accumulation of 18F-FDG within the pancreatic tumour and clear visualization of the tumour on the MRI images. Delineation of the tumours was aided by superimposition of the images from the two imaging methods, which was achieved by using a system of surface markers. PMID- 7478397 TI - Dual-tracer autoradiography using 125I-iomazenil and 99Tcm-HMPAO in experimental brain ischaemia. AB - To investigate the utility of neuroreceptor imaging in ischaemic cerebrovascular disorders, dual-tracer autoradiography using 99Tcm-hexamethylpropyleneamine oxime (HMPAO) for the evaluation of cerebral blood flow and 125I-iomazenil for the evaluation of central-type benzodiazepine receptor density was performed in experimental brain ischaemia created by occlusion of the unilateral middle cerebral artery of the rat. In the acute phase of ischaemia, 125I-iomazenil accumulation showed less of a decrease than 99Tcm-HMPAO in the cerebral cortex of the lateral convexity and in the lateral segment of the caudate putamen in the lesioned cerebral hemisphere. In the sub-acute phase, both 125I-iomazenil and 99Tcm-HMPAO accumulation increased in the lesion compared with the acute phase. A large accumulation of 99Tcm-HMPAO and 125I-iomazenil in the lesion was considered to be due to luxury perfusion and penetration of 125I-iomazenil hydrophilic metabolites from the blood into the brain tissue through the altered blood-brain barrier. In the chronic phase, 125I-iomazenil accumulation showed a more marked decrease than 99Tcm-HMPAO in the lesion. Moreover, the ipsilateral thalamus, which is remote from the lesion, revealed decreased 125I-iomazenil accumulation despite normal 99Tcm-HMPAO accumulation. Since the central-type benzodiazepine receptors are principally located not on the glial cells but on the neurons, the receptor density may exhibit a change that parallels the neuron density. These results suggest that central-type benzodiazepine receptor imaging is useful for the evaluation of neuronal damage when used in conjunction with brain perfusion imaging in ischaemic cerebrovascular disorders, except in the sub-acute phase. PMID- 7478398 TI - Inter-ictal brain SPET in frontal epilepsy: correlations with stereo electroencephalography. AB - Single photon emission tomography (SPET) imaging holds promise for localization of the site of extratemporal seizures, but limited data currently exist; in particular, correlations with stereo-electroencephalography (S-EEG) have not been made. Ten patients aged 14-44 years (mean 25 years) with a proven frontal or central epilepsy by S-EEG and post-surgical follow-up were studied retrospectively: 7 patients had frontal cortectomy and one patient had a callosotomy for bifrontal epilepsy. All patients underwent clinical, inter-ictal and ictal video-EEG, computed tomography scan and/or magnetic resonance imaging, SPET and S-EEG examinations. SPET was performed inter-ictally, while on usual epileptic medications, using 99Tcm-HMPAO (n = 4) or 123I-IMP (n = 6) as the perfusion tracer. The SPET images were evaluated independently by two observers, blind to any data other than the diagnosis of frontral or central epilepsy. Localization of inter-ictal SPET hypoperfusion was compared with the epileptogenic (EZ), irritative (IZ) and lesional (LZ) zones, as defined by S-EEG. Six patients showed structural frontal abnormalities. One patient had normal SPET and one had a contralateral hypoperfusion. Therefore, concordance of sides was found in 8 of 10 patients (including one with bilateral SPET and S-EEG abnormalities). The hypoperfusion was equal to or larger than the EZ + IZ + LZ in 6 patients (5 had a frontal lesion). SPET hypoperfusion was smaller than the EZ in one patient, and different from the EZ, IZ and LZ in two patients. Although this was a retrospective study, it provides qualitative data regarding the significance of inter-ictal SPET abnormalities in frontal or central epilepsy.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7478399 TI - Imaging infection and inflammation in an African environment: comparison of 99Tcm HMPAO-labelled leukocytes and 67Ga-citrate. AB - A considerable segment of the population of South Africa has a disease spectrum similar to those of the industrialized countries. However, a much larger proportion of its inhabitants comes from historically disadvantaged communities and is plagued by diseases typical of the developing world. Infectious diseases head the list as a cause of mortality in certain age groups. We have studied 108 patients presenting with a wide array of infections. Fifty-eight patients were examined with 67Ga-citrate and with 99Tcm-HMPAO-labelled leukocytes, 40 with leukocytes alone and 10 who had a high pre-test probability of tuberculosis with 67Ga only. The sensitivity and specificity of 99Tcm-labelled leukocyte scintigraphy in the 58 patients who had both studies were 89 and 84%, respectively, while the corresponding values for 67Ga scintigraphy were 81 and 74%, respectively. The overall sensitivity and specificity for 99Tcm-labelled leukocytes for detecting inflammatory sites in all 98 cases were 92 and 89%, respectively. Although labelled leukocytes were the better of the two agents for the early diagnosis of infections with a high probability of neutrophil infiltration, 67Ga remains an excellent alternative. It is the first choice in patients in whom tuberculosis or a neoplastic process is suspected as a cause of fever. It is also safer to use in areas where labelling of leukocytes is inadvisable because of inadequate sterility or where the risk is high of infecting other patients or staff with HIV. PMID- 7478400 TI - 201T1 and 99Tcm-J001X macrophage scintigraphy: two radionuclide imaging techniques for the surveillance of acute localized radiation over-exposures. AB - The therapeutic or surgical management of acute localized irradiation is very complicated due to the delayed occurrence of ionizing radiation effects. There is a great need for non-invasive imaging techniques to delineate healthy from exposed tissues. Such a technique should be of value considering that spread of damage can occur from apparently silent fibrotic foci. Of the scintigraphic procedures, 99Tcm first-pass scintigraphy has already been recommended by the International Atomic Energy Agency (IAEA) for the evaluation of irradiated tissues. In order to improve the scintigraphic surveillance of accidental over exposures, the potentials of 201T1 and 99Tcm-J001X were evaluated on an experimental porcine model reproducing the late fibrotic effect encountered after localized irradiation. J001X, an acylated poly-(1,3)-galactoside of bacterial origin, was used for the functional imaging of the inflammatory reaction which took place after irradiation. This scintigraphy, based on targeting of activated macrophages recruited by lesions, was performed together with 201T1 scans for the assessment of blood perfusion and cellular metabolism. Our results demonstrate that these two radiopharmaceuticals provide reliable information for the management of accidental localized over-exposure, J001X allowing the assessment of the inflammatory reaction and 201T1-chloride being mainly useful for imaging the delayed onset of fibrosis. PMID- 7478401 TI - In vitro evaluation of neutrophil viability after exposure to a hypotonic medium. AB - Separation techniques for radiolabelled leukocytes have inherent problems with contaminants (e.g. platelets and erythrocytes). Hypotonic lysis methods can eliminate the erythrocytes, but the question of neutrophil viability after an exposure to a hypotonic solution (i.e. sterile water) remains. Ficoll/ hypaque two-density gradient separation was performed on donor whole blood to obtain a pure neutrophil suspension. A timed sequence of water exposure was done for 5-100 s on the neutrophil preparations. The viability of these preparations was evaluated using flow cytometry and chemotaxis. The trypan blue staining method was used to document cell death. With water exposures ranging up to 100 s, 2.04 +/- 1.80% neutrophils exhibited cellular degradation by flow cytometry, and all samples demonstrated viable neutrophils by chemotaxis and trypan blue staining. The hypotonic medium exposure times for leukocyte separations should be less than 30 s for neutrophils to retain their viability by these in vitro techniques. PMID- 7478402 TI - Perfusion lung scanning in pulmonary hypertension. PMID- 7478403 TI - Radiopharmaceutical maladministration: what action is required? PMID- 7478404 TI - Radiolabelled peptides: now and the future. AB - Designed by nature for stimulating, inhibiting or regulating numerous life functions, for a long time peptides have been considered ideal agents for therapeutic applications. In recent years, radiolabelled peptides have emerged as a new, useful class of radiopharmaceuticals for diagnosis of a variety of endocrine tumours. Although clinical results with 111In- and 125I-labelled peptides have been excellent, extensive efforts have been made in preparing and evaluating peptides labelled with 99Tcm. This is the raidonuclide of choice by virtue of its cost-effectiveness, availability and desirable nuclear characteristics. With 99Tcm, certain peptides can be labelled 'directly' without a loss of functional specificity, but certain peptides must be labelled using bifunctional chelating agents. The bifunctional chelating agent methods are chemically complex and frequently inefficient. Instant kit preparations are, however, possible. Important tests in the evaluation of these agents are receptor binding assays as well as examinations in experimental pre-clinical models. Peptides are small and can be taken up in target tissues non-specifically by increased capillary permeability. However, something that is frequently forgotten during such evaluations are control studies with non-specific peptides of similar size and charge. Radiolabelled peptides aimed at instant in vivo interaction with neutrophils or platelets must also be examined carefully to ensure that they do not induce neutropaenia or thrombocytopaenia. The preparation of radiolabelled peptides is not only challenging but also exciting, since these agents will continue to be promising and rewarding radiopharmaceuticals for decades to come. PMID- 7478405 TI - Evaluation of 99Tcm-tetrofosmin as a myocardial perfusion agent in routine clinical use. AB - Recent trials with selected patients have indicated that 99Tcm-tetrofosmin is a suitable agent for myocardial perfusion imaging. We performed 99Tcm-tetrofosmin perfusion imaging in an unselected group of 297 patients routinely referred to our department. Single photon emission tomographic (SPET) imaging was performed 45-60 min post-injection using a 2-day stress and rest protocol. Altogether, 192 patients were stressed on a treadmill and 105 using intravenous dobutamine. Comparison with angiography was possible in 86 patients, 65 of whom had atheromatous coronary artery disease and 21 of whom had normal coronary arteries (6 of whom fulfilled the criteria for syndrome X). The sensitivity for the detection of coronary artery disease was 94% (93% for exercise stress and 95% for dobutamine). The overall specificity was 85% (87% for exercise stress and 80% for dobutamine in the 15 normal patients and the segments supplied by disease-free coronary vessels in patients with disease elsewhere). We conclude that 99Tcm tetrofosmin is a highly sensitive and specific agent for the detection of coronary artery disease, using both exercise and dobutamine stress, with few limitations. PMID- 7478406 TI - Inhalation of pertechnegas: similar clearance from the lungs to that of inhaled pertechnetate aerosol. AB - It has been reported that the inhalation of pertechnegas (TGO3) generated under an atmosphere of 3% oxygen (O2) in argon (Ar) instead of 100% Ar without O2 in the technegas generator, offers a simple but accurate quantification of clearance from the lungs, permitting it to be used in place of 99Tcm-diethylenetriamine pentaacetic acid (DTPA). The disappearance from the lungs of inhaled TGO3 was so similar to that of inhaled pertechnetate aerosol (TcO4) in this study, that there was neither a statistically significant difference in the clearance half-time (t1/2), nor a difference in the lung images between them. Neither TGO3 nor TcO4 inhalation could distinguish smokers from non-smokers using t1/2. When inhaled, TGO3 appears to behave in a similar manner to TcO4 in the lungs and hence cannot be used in place of 99Tcm-DTPA in clinical practice. PMID- 7478407 TI - Diagnostic value of 169Yb-cis-dichlorodimethionine platinum scintigraphy in patients with malignant lymphomas. AB - Ytterbium-169 (169Yb) cis-dichlorodimethionine platinum (169Yb-PtCl2Meth2) is a new agent of low toxicity with an affinity for neoplastic tumours. The aim of this study was to assess the diagnostic value of 169Yb-PtCl2Meth2 scintigraphy in patients with malignant lymphomas. 169Yb-PtCl2Meth2 scintigraphy was performed in 67 patients with histologically proven malignant lymphoma. Liver, spleen, kidney and bone were among the normal tissues that showed uptake of the radiopharmaceutical. The compound is excreted in the main through the urinary tract. There was no significant activity in the bowels. For evaluation of the images, uptake of activity was scored according to a 5-point scale. In total, 888 sites were evaluated. The sensitivity, specificity and accuracy of lymphoma detection for the body as a whole was 80, 89 and 87%, respectively. The best results were obtained for the head (sensitivity 91%, specificity 89%), neck (sensitivity 84%, specificity 90%) and chest (sensitivity 82%, specificity 84%). In the sub-diaphragmatic region, sensitivity and specificity were 76 and 84%, respectively. Scintigraphy with 169Yb-PtCl2Meth2 appears to be a sensitive, non invasive procedure for the staging of malignant lymphomas. The results suggest that it is possible to monitor the therapy of malignant lymphoma by means of 169Yb-PtCl2Meth2 scanning. PMID- 7478408 TI - Somatostatin receptor imaging in CNS tumours using 111In-octreotide. AB - This study evaluates the in vivo visualization of somatostatin (SS) receptors in central nervous system (CNS) tumours using 111In-octreotide imaging and discusses the clinical implications. Ninety-five patients with histologically confirmed diagnosis of CNS tumours were imaged 2-4 and 24 h after the intravenous injection of 111-185 MBq of 111In-octreotide. An uptake index was computed using tumour/non tumour ratios evaluated using a standard region-of-interest method. Semi quantitative immunohistochemical studies of SS binding sites were performed on frozen tumour sections. All meningiomas, most pituitary adenomas and many glial tumours showed a positive scan, whereas all neurinomas, craniopharingiomas and ependymomas had negative receptor scans. Radio-octreotide uptake varied among the SS receptor positive CNS tumours: very intense in meningioma, intermediate in pituitary adenoma and of a low grade in glioma. The results of immunohistochemical studies confirmed the scintigraphic findings in all cases. We believe 111In-octreotide is a suitable radiopharmaceutical for characterizing CNS tumours in vivo as SS receptor positive or negative. This new neuronuclear imaging technique may be useful for differential diagnosis in selected cases, for post-surgical follow-up and in the assessment of differentiation in glial tumours. PMID- 7478410 TI - A simple method for estimating the skeletal uptake of diphosphonates. AB - It is possible to obtain an estimate of the bone uptake of diphosphonates after 3 h (BU) on the basis of the difference between the dose administered and the sum of the component eliminated in the urine plus the component present in the extra osseous volume of distribution. The results of this simple method have been compared in 15 patients using whole body retention at 24 h (determined by means of a complete urine collection) and the ratio of plasma concentrations of 51Cr EDTA/99Tcm-HMDP after 3 h (DPS). Both DPS and BU can be used on a practical basis, and therefore be used routinely with scintigraphy. The added advantage is that, for BU, the use of a second radiopharmaceutical is avoided. PMID- 7478409 TI - Renal excretion of meta-iodobenzylguanidine after therapeutic doses in cancer patients and its relation to dose and creatinine clearance. AB - Iodine-131 radioiodinated meta-iodobenzylguanidine (131I-MIBG) is used for diagnostic scintigraphy and radionuclide therapy of neural crest-derived tumours. In higher doses (up to 80 mg), non-radioactive MIBG is now evaluated for palliation in carcinoid patients. After administration of therapeutic doses of 131I-MIBG (3.7-7.4 GBq, 1.7-5.8 mg MIBG) to patients aged 2-73 years, 53 +/- 8.8%, 69 +/- 7.8% and 83 +/- 7.0% of the dose was cumulatively excreted as MIBG in the urine after 24, 48 and 72 h, respectively. Within the MIBG dose range of 1.7-80.0 mg, a linear relationship was found between the excretion rate over the first 24 h (mg per 24 h) and the dose. In adults, the MIBG excretion rate over the first 24 h (% of dose per 24 h) was shown to be only partially related to the glomerular filtration rate (GFR). PMID- 7478411 TI - The statistical power of [15O]-water PET activation studies of cognitive processes. AB - [15O]-Water positron emission tomography (PET) scanning is an important means of investigating the neural correlates of cognitive processes. This study analyses the statistical power of these experiments under usual experimental conditions. Eleven subjects were scanned repeatedly while they undertook an active-baseline task. Using an empirically derived estimate of variance, the statistical power to detect task-induced increases in regional cerebral blood flow was calculated for different values of magnitude of change, number of subjects and probability of Type I error. The results (a) provide an empirical basis for selecting the number of subjects for cognitive studies using [15O]-water PET; (b) illustrate the importance of considering the statistical power of the test, while making a physiological interpretation of the results; and (c) highlight the problems in concluding the non-involvement of a given brain region in a cognitive task, based solely on the results of a PET study. PMID- 7478412 TI - Hypertrophic pulmonary osteoarthropathy in nasopharyngeal carcinoma: an early sign of pulmonary metastasis. AB - The aims of this study were to determine the incidence of hypertrophic pulmonary osteoarthropathy (HPO) in nasopharyngeal carcinoma (NPC) and assess its clinical significance. Altogether, 407 NPC patients were reviewed retrospectively. HPO was identified by 99Tcm-methylene diphosphonate bone scans and related clinical and radiographic evidence. Pulmonary metastases, bony metastases and titre of anti Epstein Barr virus (EBV) immunoglobulin were assessed in patients with and without HPO. The patients had a mean (+/- S.D.) age of 50.4 +/- 12.4 (range 17 73) years. HPO was found in 27 of the 407 (6.6%) NPC patients, among whom 13 (48%) had pulmonary metastases. HPO preceded lung metastases by 7-22 months (14.4 +/- 6 months) in 7 (52%) patients. Six patients had overt lung metastases at the time of the bone scan. No significant difference was found in anti-EBV immunoglobulins between the patients with or without HPO, nor in the incidence of bony metastases between these two groups of patients. HPO should be regarded as an early sign of pulmonary metastases. PMID- 7478414 TI - Surveying the obstetric and gynecologic literature: making epidemiology and biostatistics work for you. PMID- 7478413 TI - Effects of oral erythromycin on upper gastrointestinal motility in patients with non-insulin-dependent diabetes mellitus. AB - Twenty patients (18 males, 2 females) with type II (non-insulin-dependent) diabetes mellitus were enrolled in the study. They were aged 49-72 years (mean age 65 years). Radionuclide-labelled water was used to measure oesophageal motility, expressed as the oesophageal mean transit time (MTT). A radionuclide labelled solid meal was used to measure gastric motility, expressed as the half time of gastric emptying (T1/2GET). A baseline study was performed before oral erythromycin therapy. After a 2-week course of treatment, the subjects underwent a second study. Fasting blood sugar (FBS) was also monitored in each study. In the baseline study, MTT was 8.88 +/- 2.00 s and T1/2GET was 198.0 +/- 58.9 min. After treatment with erythromycin, MTT decreased to 7.48 +/- 2.24 s (P < 0.01) and T1/2GET decreased to 137.1 +/- 71.2 min (P < 0.01). In addition, the FBS decreased from 159.0 +/- 40.2 mg dl-1 at baseline to 139.2 +/- 39.8 mg dl-1 after 2 weeks of erythromycin treatment (P < 0.05). We conclude that erythromycin is an effective prokinetic agent for diabetic gastroparesis, and that improved oesophageal transit and gastric emptying may improve glycaemic control. PMID- 7478416 TI - Vulvar pain syndrome: a review. AB - Vulvar pain syndrome (vulvar vestibulitis, vulvodynia, burning vulvar syndrome) was first described at the end of the last century. Although more than 100 years have passed, the cause(s) of the disorder remains elusive. This review of the literature endeavors to collate the known facts relating to vulvar pain syndrome and to expose those hypothetical etiologies which fall short of establishing any scientific foundation. The diagnosis of vulvar pain syndrome is established on the basis of historical data which detail a rather abrupt onset of vestibular itching, burning, dryness which may be intermittent or continuous. Commonly, patients present with a history of chronic treatment for recurrent fungal infections. Sexual relations become uncomfortable or intolerable. Unanimously, the patients describe a panorama of topical creams and ointments which fail to alleviate the symptoms. Physical examination typically verifies hyperesthesia of the vestibular skin when touched lightly with a cotton-tipped applicator. Similarly, the vestibular skin shows varying degrees of redness. The treatment programs for vulvar pain syndrome are as diverse as the multitude of etiological hypotheses. This review includes data pointing out those therapeutic measures that appear valueless and that may add to the patient's chronic discomfort. PMID- 7478415 TI - Umbilical cord blood for use in transplantation. AB - Bone marrow transplantation has rapidly progressed over the last two decades offering cure and prolonged disease-free survival for patients suffering from certain hemato-oncological malignancies or metabolic disorders. However, bone marrow transplantation is limited by the paucity of major histocompatibility loci antigen (HLA)-matched donors, and the morbidity and mortality due to graft-versus host disease. Recently it has been shown, that umbilical cord blood represents a unique source of transplantable hematopoietic progenitor cells. Currently, human umbilical cord blood from a newborn sibling has been used successfully for hematopoietic reconstitution of approximately 40 children with congenital or malignant diseases. Establishment of umbilical cord blood banks might alleviate some of the problems associated with bone marrow transplantation. The developments in this field which have occurred during the last decade, as well as the importance of cooperation between the obstetric and transplantation staff, are discussed in this review. Certain ethical problems remain surrounding the issue of using human umbilical cord blood for allogeneic transplantation which must be addressed. PMID- 7478417 TI - Ascites due to endometriosis. AB - Ascites is a rare but important complication of endometriosis because it mimics ovarian cancer. Most cases occur in nulliparous young black women and present with massive ascites. Treatment is effected by ablation of ovarian function by surgery, radiotherapy, or suppression of endometriosis by endocrine therapy. The pathogenesis is unknown. In this paper, we present a case report and review of the other 19 cases in the literature. Because of the age of most of these women, endocrine therapy is preferred rather than castration. The majority of symptoms and signs of endometriosis are well known, including pelvic pain, dysmenorrhea, dyspareunia, infertility, and pelvic tenderness with or without masses. However, it is seldom appreciated that the disease can be a cause of, and can present with ascites, often massive and recurrent. It is important for gynecologists and oncologists to be aware of this entity because the presence of ascites with abdominal and/or pelvic masses and weight loss immediately suggests the diagnosis of malignancy, and the possibility of endometriosis is rarely considered. We are reporting a case of endometriosis causing massive and recurrent ascites, along with a review of the literature and a discussion of the epidemiology, pathogenesis, and management of this disorder. PMID- 7478418 TI - Teaching basic surgical skills in Ob-Gyn. PMID- 7478419 TI - Lower urinary tract dysfunction in pregnancy: a review. AB - It has long been observed that pregnancy may influence the development and course of urinary tract disorders. The physiological and anatomical changes inherent in normal pregnancy and the changing hormonal environment are generally assumed to play a role in the pathogenesis of urinary tract symptomatology. The purpose of this review is to examine the reported effect(s) of pregnancy on the lower urinary tract and to evaluate the possible role of pregnancy and delivery in lower urinary tract dysfunction. PMID- 7478420 TI - Fetal porencephaly: a review of etiology, diagnosis, and prognosis. AB - Porencephaly may result from either a developmental anomaly or an antepartum intraparenchymal insult. Pre- and postnatal ultrasonography can demonstrate characteristic images of porencephaly. There is minimal literature on diagnosis and outcome of antenatally diagnosed porencephaly. There are two types of porencephaly: type I is generally due to an antepartum intraparenchymal hemorrhage. Type II lesions are usually developmental anomalies. Prognosis generally depends on the extent of the lesion. PMID- 7478421 TI - Angiogenesis in the human female reproductive tract. PMID- 7478422 TI - Safety, health, and ISO 9000. PMID- 7478423 TI - Stopping the burn. PMID- 7478424 TI - Health and safety in the future: the basics, improved. PMID- 7478425 TI - Special product focus: Respiratory protection. PMID- 7478426 TI - Mitoxantrone, fluorouracil plus L-leucovorin, and vinorelbine in pretreated advanced breast cancer. AB - The combination chemotherapy including mitoxantrone, fluorouracil and leucovorin has proven to be effective and well-tolerated in advanced breast cancer (ABC). No data are available on the association with navelbine, a new vinka alkaloid, which has demonstrated high activity in ABC. The trial was designed to evaluate feasibility and efficacy of the association of vinorelbine to mitoxantrone, fluorouracil and L-leucovorin in patients who failed a previous regimen for ABC or who relapsed within 6 months of adjuvant chemotherapy. The schedule was as follows: mitoxantrone 6 mg/m2 days 1 and 8; L-leucovorin 250 mg/m2 days 1 and 8, fluorouracil 600 mg/m2 days 1 and 8, vinorelbine 25 mg/m2 days 1 and 8, cycles being repeated every 21 days. Twenty-five patients were enrolled and are evaluable for response and side effects. One hundred cycles of therapy have been delivered (median/patient, 4 cycles). In 41 cycles a delay was required and in 38 cycles it was necessary to administer granulocyte colony-stimulating factor for neutropenia. Seven partial remissions (28%; 95% confidence interval, 12-49%), 10 stabilizations of disease and 8 progressions were observed. Although grade 4 neutropenia was observed in 44% of the patients, no grade 3-4 infections were observed. Nonhematologic toxicities were mild or moderate and included alopecia, mucositis and phlebitis. In conclusion, the schedule employed, although correlated with a moderate activity, does not seem to be superior to other regimens with mitoxantrone, fluorouracil, and leucovorin. PMID- 7478427 TI - Prediction of recurrence in operable breast cancer by postoperative changes in prolactin secretion. AB - It has been demonstrated that breast surgery may induce prolactin (PRL) increase. Because of the potential stimulatory role of PRL on breast cancer cells, its postoperative increase may influence the prognosis of breast cancer patients. This study was performed to evaluate the influence of surgery-induced hyperprolactinemia on recurrence rate in operable breast cancer. The study included 250 consecutive breast cancer patients, clinical stage T1-3 N0-2M0, who were observed for a median follow-up of 72 months. Surgery-induced hyperprolactinemia occurred in 108/250 patients (43%). Irrespectively of node involvement, hormonal receptor, type of surgery and adjuvant therapies, the relapse rate was significantly higher in patients who had no surgery-induced hyperprolactinemia than in those with postoperative PRL increase (64/142 vs. 23/108; p < 0.001). This difference was also significant in relation to node status (N0:22/63 vs. 5/56, p < 0.001; N+:42/79 vs. 18/52, p < 0.05). The present study shows that a surgery-induced rise of PRL, despite its potential stimulation of cancer cell growth, is paradoxically associated with a longer disease-free survival in operable breast carcinoma in both patients with or without axillary node involvement. Moreover, this study suggests that the prognosis of node negative patients who did not show postoperative hyperprolactinemia tends to be similar to that of patients with node involvement and surgery-induced PRL enhancement. Therefore, the lack of surgery-induced hyperprolactinemia would have to be grouped together with the unfavorable prognostic factors of breast cancer. PMID- 7478428 TI - Treatment failure following sequential thermoradiotherapy of locally advanced breast carcinoma occurs primarily in poorly vascularized tumors. AB - Sequential thermoradiotherapy was given to 24 patients with locally advanced breast carcinoma. The outcome of treatment was analyzed in relation to the thermal dose as well as clinical stage, tumor volume, vascular density and necrotic fraction prior to treatment. Complete or partial response was seen in all patients. Three of the patients showed local recurrence following treatment. One of the recurrences was due to a geographic miss of the radiation therapy whereas the other two recurrences were 'true' local treatment failures. Local treatment failure could not be attributed to an advanced clinical stage, large tumor masses or poor tumor heating. The recurrent tumors had a low vascular density and significant necrotic fraction prior to treatment, suggesting that treatment failure was due to the presence of chronic hypoxia. The radiotherapeutic problem of treatment resistance caused by chronically hypoxic cells is probably not eliminated with sequential thermoradiotherapy. PMID- 7478429 TI - Five distinct deleted regions on chromosome 17 defining different subsets of human primary breast tumors. AB - In this study, we analyzed 105 paired sporadic primary breast tumor and normal tissue samples for loss of heterozygosity (LOH) on chromosome 17, using 12 polymorphic markers. We have identified partial or interstitial LOH in five separate regions of chromosome 17. Two of the deleted regions lie on the short arm of the chromosome, the first (region I, D17S5) in the telomeric part, distal to TP53 and the second spanning the TP53 gene (region II). Three of the five deleted regions lie on the long arm of chromosome 17: region III, on the proximal long arm between D17S250 and THRA1; region IV, between D17S776 and D17S579, including the BRCA1 gene, and region V, located distal to D17S733. No statistically significant correlations were observed between clinicopathological characteristics or steroid hormone receptor status and deletion of either region I or II. However, patients whose tumors had LOH for region I showed relapse or death more frequently than patients with tumors informative for this region but without LOH (p = 0.002). Statistically significant correlations between LOH at each of the three deleted regions of 17q and a high mitotic index were observed (region III, p = 0.005; region IV, p = 0.02, and region V, p = 0.004). In addition, LOH at region IV showed a significant association with paucity of estrogen receptors (p = 0.01). Our results show a complex pattern of LOH on chromosome 17 in breast cancer and a correlation of these events with different clinical parameters. This pattern suggests that particular subsets of allele loss may contribute specifically to different clinically defined subsets of sporadic breast tumors. PMID- 7478430 TI - Activity of formestane in de novo tamoxifen-resistant patients with metastatic breast cancer. AB - In order to assess the feasibility of a sequential hormonal treatment after tamoxifen failure, 24 postmenopausal advanced breast cancer patients (median age 60 years; ECOG PS < or = 1) were treated with formestane (4 hydroxyandrostenedione) 250 mg i.m. fortnightly; 19 patients were estrogen receptor-positive. The sites of metastatic disease were soft tissue in 22 patients, viscera in 9 and bone in 18. The patients were considered evaluable for tumor response after four doses of formestane. Objective responses were observed in 8/24 patients (33%) with one complete and seven partial responses. The median response duration was 9.5 months. The complete response was obtained on skin. We conclude that although the number of complete responses appears to be unsatisfactory, de novo tamoxifen-resistant breast cancer patients are suitable for further hormonal treatment with formestane. PMID- 7478431 TI - Effects of eicosapentaenoic and docosahexaenoic acid on cell growth and prostaglandin E and leukotriene B production by a human breast cancer cell line (MDA-MB-231). AB - We investigated the in vitro effects of eicosapentaenoic acid (EPA) and docosahexaenoic acid (DHA), with or without the addition of linoleic acid (LA), on cell growth and prostaglandin E (PGE) and leukotriene B (LTB) secretion by a human breast cancer cell line (MDA-MB-231). With or without the addition of LA, EPA and DHA suppressed cell growth and thymidine incorporation and reduced the secretion of PGE and LTB. In a univariate analysis, cell growth was significantly associated with both LTB and PGE concentrations when cells were treated with DHA or EPA, independent of the addition of LA. However, multivariate regression analysis showed that cell growth was more closely associated with the PGE concentration rather than the LTB concentration. These data suggest that both EPA and DHA suppress cell proliferation in the MDA-MB-231 cell line by inhibition of the cyclooxygenase rather than the lipoxygenase pathways. However, the exact mechanism underlying the antitumor activity of EPA and DHA remains unclear. PMID- 7478432 TI - Influence of acetylsalicylic acid and metabolites on DU-145 prostatic cancer cell proliferation. AB - Conflicting reports have been published on the anti-tumour activities of acetylsalicylic acid in various cancers. Therefore, the effect of acetylsalicylic acid and its major metabolites has been studied on human prostatic carcinoma DU 145 cells. Investigations concentrated on the influence of acetylsalicylic acid, salicylic acid and salicyluric acid, on cell proliferation, DNA- and protein synthesis of DU-145 cells. DNA and protein synthesis determinations were done in vitro by [3H]thymidine and [3H]glycine incorporation, respectively. No effect on cell plating efficiency was observed, however proliferation studies showed that acetylsalicylic acid and salicylic acid inhibited cell growth (10 mM, 100% inhibition). No significant effect on cell proliferation was ascertained with salicyluric acid. Both DNA and protein synthesis were 40% inhibited by 0.1 mM acetylsalicylic acid. This study demonstrates that acetylsalicylic acid exhibits a significant influence on cell growth of prostatic DU-145 cells. These preliminary results may contribute to a better understanding of the anti-tumour capabilities of acetylsalicylic acid. PMID- 7478433 TI - Second-line treatment with high-dose 5-fluorouracil and folinic acid in advanced colorectal cancer refractory to standard-dose 5-fluorouracil treatment. AB - Results with second-line treatment of advanced colorectal cancer are poor. Preliminary phase I/II results suggest encouraging response rates to high-dose 5 fluorouracil (FU) (2,600 mg/m2) combined with folinic acid (FA) in pretreated patients with advanced colorectal cancer. To determine the significance of weekly high-dose FU/FA in colorectal cancer refractory to weekly standard doses of FU/FA, a phase II study was initiated. 69 patients with metastatic colorectal cancer refractory to first-line weekly standard-dose FU/FA were treated with weekly high-dose FU 2,600 mg/m2 combined with FA 500 mg/m2 as a 2-hour infusion prior to FU application. Pretreatment was highly homogenous and consisted of weekly FU 500 mg/m2 combined with FA 500 mg/m2 or FA 20 mg/m2 or the pure stereoisomer of FA (6S-FA) 250 mg/m2. At the time of disease progression under first-line therapy, patients were transferred to high-dose FU/FA within 4 weeks. Treatment was continued until tumor progression under therapy was documented. Of the 69 evaluable patients, 17 (24.6%) achieved partial response (PR), 42 (60.9%) had no change (NC) and 10 (14.5%) had progressive disease under therapy. The median duration of PR was 7 months, the median time to progression of NC was 4 months. Median survival of all patients was 9 months, of patients with PR, 11.5 months. 33/38 patients with tumor-related pain experienced impressive relief under therapy. Prognostic factors for a beneficial outcome were complete response/PR under first-line therapy, a small number of metastatic sites and a good Karnofsky performance status. Moderate toxicity was observed, and the pattern of toxic events and severity did not differ from standard-dose FU regimens.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7478434 TI - Prognostic evaluation of curatively resected locally advanced gastric cancer patients with preoperative downstaging chemotherapy assessed by histochemical and pharmacologic means. AB - The aim of the present study was to investigate whether the rate of thymidylate synthetase inhibition (TSIR) and the rate of proliferating cell nuclear antigen expression (PCNA-R) in gastric cancer tissues, which can be obtained within a short period after surgery, were predictive and quantitative prognostic factors for locally advanced gastric cancer patients with preoperative down-staging chemotherapy. Curatively resected 30 locally advanced gastric cancer patients with preoperative chemotherapies were studied. Three-year survival analysis showed that the higher TSIR and the lower PCNA-R significantly predicted better prognosis (p < 0.01 and p < 0.05, respectively). Multiple regression test showed that the TSIR was a significantly predictive variable for 1-year survival (p < 0.05). The TSIR and PCNA-R could be predictive and quantitative prognostic factors in advanced gastric cancer patients who received preoperative downstaging chemotherapy. PMID- 7478435 TI - Prognostic value of combination assays for CEA and CA 19-9 in gastric cancer. AB - Preoperative serum CEA and CA 19-9 levels in 158 patients with gastric cancer were analyzed with respect to prognostic factors, using univariate and multivariate analysis. The incidence of high preoperative levels of both CEA and CA 19-9 was 10.1% (16/158). 13.9% (22/158) showed high CEA levels and normal CA 19-9 levels, whereas the reverse was true in 16.5% (26/158). Neither marker showed a high level in 59.5% (94/158). The multivariate analysis showed that in addition to tumor stage, the depth of invasion, liver metastasis and peritoneal dissemination, combination assays of preoperative serum CEA and CA 19-9 levels were an independent prognostic factor. Combination assays of preoperative serum CEA and CA 19-9 will allow us to conduct a more careful postoperative follow-up of high-risk patients, and also help determine the optimum adjuvant chemotherapy. PMID- 7478436 TI - 5-Fluorouracil and d,l-leucovorin calcium are active to treat unresectable hepatocellular carcinoma patients: preliminary results of a phase II study. AB - Twenty-five patients affected with unresectable hepatocellular carcinomas (HCCs) were treated with 370 mg/m2 5-fluorouracil (5-FU) plus 200 mg/m2 racemic leucovorin both for 5 consecutive days. The chemotherapy cycle was repeated every 4 weeks until disease progression. One complete remission (4%) and 6 partial responses (24%) were obtained, that is 7 objective responses in all (28%, ninety five confidence interval: 10.1-45.9%). All responders had a good PS and only 1 of them presented with bulky disease. Five patients (20%) exhibited stable disease, while the remaining 13 patients (52%) progressed. Toxicity was mild: 11 patients (44%) had grade II/III mucositis, 10 patients (40%) grade II diarrhea, 7 patients (28%) grade II nausea, 2 patients grade III granulocytopenia and 1 patient only (4%) grade I skin toxicity. This regimen made it possible to obtain a high rate of objective responses even in inoperable HCCs which are commonly considered as chemoresistant lesions. PMID- 7478437 TI - Effect of hyperthermia on intracellular pH in human U-87 MG glioblastoma cells. AB - The effect of hyperthermia at 43 degrees C on intracellular pH (pHi) in human U 87 MG glioblastoma cells was studied by using the fluorescent probe 2',7' bis(carboxyethyl)-5(6)-carboxyfluorescein-pentaacetoxymethyl ester. The presence of Na+/H+ antiporter activity in the cells were demonstrated by the Na(+) dependent increase in intracellular pH (pHi) after cellular acidification in the absence of HCO3-. Hyperthermia at 43 degrees C caused significant decrease in pHi. The acidification was readily reversible by cooling the cells back down to 37 degrees C. The pHi change was inhibited by the addition of 1 mM amiloride in the incubation medium. Amiloride and hyperthermia exhibited a synergistic effect in suppressing thymidine incorporation into the cells. PMID- 7478438 TI - Association between orosomucoid types and cancer. AB - Orosomucoid (ORM) is a polymorphic acute-phase reactant with immunosuppressive properties. Previous investigations have suggested that ORM and other acute-phase reactants may act as blocking factors protecting tumor cells against immunological attack, thereby contributing to the 'immune escape' of the tumor. ORM types were studied by isoelectric focusing and immunoprinting in patients with carcinoma of the breast, lung, ovary and endometrium and in population controls. In accordance with a previous study, the frequencies of the ORM1*2 allele and the ORM1 2 phenotype were found to be significantly increased in patients with different types of carcinomas. The results suggest that the ORM1 2 variant may influence the progression of cancer by being more immunosuppressive than the ORM1 1 variant. The relative risk for carcinoma in patients with the ORM1 2 type was 1.56 (95% confidence interval:1.16-2.09). PMID- 7478439 TI - Primary T cell leptomeningeal lymphoma--successful treatment with systemic chemotherapy. AB - Primary central nervous system lymphoma (PCNSL) is rare, accounting for only 1-2% of non-Hodgkin's lymphoma, and primary isolated leptomeningeal lymphoma is even rarer. It may create a diagnostic problem, particularly when the tumor cells are of T cell lineage. We herein report a patient with primary T cell leptomeningeal lymphoma. The final diagnosis was confirmed by the cytogenetic study which revealed clonal aberration, isochromosome of long arm of chromosome 7, in the lymphoid cells from CSF. She was treated with a novel protocol of systemic chemotherapy specifically designed for the CNS lymphoma. The regimen consisted of carmustine, vincristine, high-dose methotrexate, etoposide, and methylprednisolone. A total of 5 courses were given and she was still in complete remission at the time of reporting, 21 months after the establishment of diagnosis. PMID- 7478440 TI - Acute disseminated intravascular coagulation syndrome in cancer patients. AB - Hemostatic abnormalities are rather frequent in cancer patients either in hematological or in solid tumors. Acute disseminated intravascular coagulation (DIC) is a rare coagulopathy in cancer patients, but when it develops it becomes rapidly fatal. Between June 1988 and December 1992 we observed 8 cases of acute DIC occurring in gastric cancer (4 patients), breast cancer (3 patients) and high grade non-Hodgkin lymphoma (1 patient). In 3 patients affected by gastric carcinoma, acute DIC was the first manifestation of the presence of the tumor, while in the other patients DIC occurred during the course of the disease. All the patients were treated with heparin, fresh frozen plasma and platelet support, but only in 1 patient was a short duration improvement of clinical conditions and coagulation tests recorded. Acute DIC can be the first manifestation of gastric tumors and the presence of the hemorrhagic syndrome associated with thrombocytopenia, hypofibrinogenemia and fibrin/fibrinogen degradation products should initiate a search for gastric carcinoma. PMID- 7478441 TI - Oral ondansetron and intravenous dexamethasone in the prevention of cisplatin induced emesis. A phase II trial in women. AB - Fifty-eight previously untreated patients with gynecological cancer, assigned to cisplatin-based chemotherapy (40-80 mg/m2), received the following antiemetic treatment: day 0, oral ondansetron 8 mg 3 times/day + intravenous dexamethasone 16 mg; days 1-7, oral ondansetron 8 mg twice/day. In cycle 1 complete or major control (0-2 emetic episodes) was achieved in 94.6% of the patients in the acute phase (day 0) and in 89.2% in the delayed phase (day 1-7). In the subgroup receiving cisplatin > or = 75 mg/m2 the effect on acute and delayed emesis decreased significantly with subsequent courses. Reversible side effects were observed in 8.9% of the cases. Oral ondansetron was efficacious, well tolerated and is worth testing further in randomized trials with intravenous therapy. PMID- 7478442 TI - [The cellular aspects of the development of synovial joints and articular cartilage]. AB - There have been many reports on the histological development of mammalian diarthrodial or synovial joints. While these are useful for comparative purposes, they tell us little of the cellular basis of joint morphogenesis which must underlie a number of morphogenetic defects. The process of joint morphogenesis is complex and can be subdivided into a number of facets and this report will focus on 2 of them. First, the process of joint cavitation in the chick metatarsophalangeal joint, where we propose that the selective secretion of hyaluronan into the presumptive cavity plays a central role. Secondly, the development of articular cartilage where we have used the South American opossum Monodelphis domestica as a model for mammalian development. Like most marsupials, the young are born at a much earlier developmental stage than eutherian mammals. Using antibodies which detect proliferating chondrocytes and those synthesizing insulin-like growth factors 1 and 2 and insulin-like growth factor 1 binding protein, we report that the majority of growth (as assessed by these indicators) appears appositional. PMID- 7478443 TI - [The creation of genetic cryobanks and the use of the methods of developmental biology as a means for preserving rare animal species. I. The cryopreservation of the sperm of wild mammals]. AB - A review of published data on cryoconservation of the sperm of wild animals. Progeny of females inseminated with frozen-thawed sperm was obtained in 21 species and subspecies, including 11 from the Red Data Book. Mobility of the spermatozoa after freezing-thawing was estimated in 74 species, including 13 from the Red Data Book. Studies on cryoconservation have recently been carried out predominantly on the Red Data Book species, as well as the species intensely bred in the countries, where several founder individuals were introduced (deer in Australia and New Zealand). Large zoos and some universities in the USA, Canada and Great Britain are the main research centers. PMID- 7478444 TI - [Pattern formation in the early embryogenesis of the clawed toad]. AB - Establishment of egg polarity, separation of germ layers, appearance of animal vegetal, dorsoventral and anteroposterior axis in Xenopus laevis embryos have been considered. The control of these processes by genes coding for growth factors, protooncogenes and homeobox-containing genes has also been reviewed. PMID- 7478445 TI - [The binding of exogenous DNA pRK31acZ by rabbit spermatozoa, its transfer to oocytes and expression in preimplantation embryos]. AB - It was shown that 35.3% ejaculated rabbit spermatozoa washed from seminal plasma are capable of interacting with heterogeneous DNA. The major part (85.2%) of bound DNA was located in the post-acrosomal part of the sperm head. After incubation with plasmid pRK31acZ the spermatozoa transferred it in the oocytes during in vivo fertilization, as shown by expression of reporter gene lacZ in 19.3% of preimplantation embryos. Additional treatment of the mobile spermatozoa with DMSO and heat shock raised the efficiency of exogenous DNA incorporation in the spermatozoa, as expressed in the percentage of embryos containing bacterial beta-galactosidase (61.7%). It is proposed to use the capture of heterogeneous DNA by the spermatozoa and its transfer in the oocytes for production of transgenic animals and in studies of regulation of gene expression at the early stages of embryogenesis. PMID- 7478446 TI - [An analysis of keratin expression in the cells of the retinal pigment epithelium during transdifferentiation in newts]. AB - Keratins of the cytoskeletal intermediate filaments have been identified immunohistochemically in pigment epithelium cells of the adult newt retina. We studied the expression of keratins in transdifferentiating and redifferentiating cells of the retinal pigment epithelium at different periods after removal of the retina. We have shown that the expression of keratins decreases. This appears as the decreased intensity of the immunospecific reaction soon after the operation. The latter observation has also been made immediately after artificial dissociation of the pigment epithelium cells of the retina isolated from nonoperated newt eyes. We found that the immunostaining of keratins was absent in the cells of normal and regenerating retina, as well as in retinal pigment epithelium cells before restoration of their structural and functional relationship with the retinal regenerate. The results obtained provide evidence for the inhibition of keratins expression during the transformation of retinal pigment epithelium into the retina. We proposed that there exists a mechanism responsible for gradual replacement of keratins by proteins of neurofilaments that have earlier been identified at the beginning of transdifferentiation of the retinal pigment epithelium. PMID- 7478447 TI - [A paradox: the short duration of life in the presence of long telomeres]. PMID- 7478448 TI - Effects of hyperventilation and hypoventilation on cochlear blood flow and endocochlear direct-current potential. AB - To understand the importance of oxygen transport to the inner ear tissue, we studied, in guinea pigs, the relationship between cochlear blood flow and endocochlear direct-current potential (EP) under different respiratory conditions. EP, a functional parameter of the stria vascularis, was recorded by a microelectrode inserted into the lateral wall of the chochlea. To measure the cochlear blood flow (CoBF), we employed laser Doppler flowmetry and recorded the flow with a probe placed on the same spot on the lateral wall. During 3 min of asphyxia, CoBF and systemic blood pressure showed irregular biphasic increases, while the EP decreased to reach a negative value. In the hypoxemia experiment, which was induced by stepwise reduction of the respiratory rate to 60%, increases in CoBF and blood pressure were evident during hypoventilation with an intermediate position of EP in the positive range. The mechanisms of these increases in two parameters are discussed from the viewpoints of sympathicotonic activity in the autonomic nervous system and the vasodilating action of CO2 during hypercapnia. In the hyperoxemia experiment, which was induced by stepwise increase in the respiratory rate to 140%, CoBF and blood pressure were found to decrease during hyperventilation with no significant change of EP. The decrease in blood pressure was considered to be due to the increase in intrathoracic pressure caused by the increased rate of artificial respiration. As for the concomitant decrease in CoBF, chemical regulation of PCO2 in the vascular bed of the lateral wall of the cochlea was thought to be a contributory factor. PMID- 7478449 TI - Expression of subtypes NF-L, NF-M and NF-H of neurofilament triplet proteins in the developing rat inner ear. AB - The expression of each subtype (NF-L, NF-M and NF-H) of neurofilament (NF) triplet proteins in the developing rat inner ear was analysed using the immunofluorescence method (FITC). On the 15th gestational day (GD), immunostaining of NF subtypes did not occur in the otic region. On the 18th GD, positivity for NF-L and NF-M occurred in nerve fibres in both the sensory and the subepithelial layers of the inner ear but was lacking for NF-H. At 4 days after birth immunoreactivity was present for each NF subtype but with topographic differences in their distribution. The cell bodies of (different?) subpopulations of vestibular and spiral ganglion cells were immunostained with monoclonal antibodies for NF-L and NF-M, but a subpopulation of both vestibular and spiral ganglion cells showed immunoreactivity for all three NF subtypes. The vestibular nerve fibres and ganglion cells evidenced stronger positivity than cochlear nerve fibres and spiral ganglion cells. This study confirms that the cytoskeletal maturation of inner ear nerve fibres precedes the onset of at least the auditory function by several days. PMID- 7478450 TI - Ultrastructure of the vestibular dark cell area in patients with acoustic neurinoma. AB - The human vestibular dark cell (DC) areas of the utricle and ampulla of the lateral semicircular canal were investigated ultrastructurally in 7 patients with acoustic neurinoma. Two types of DCs mainly constituted the epithelial cells of the DC area. One type of DC had basolateral infoldings that were closely interwoven with the melanocyte processes, and the pinocytotic vesicles were frequently found within the basolateral infoldings and the melanocyte processes. The other type of DC had basolateral infolsings stacked upon each other. The former type of DC was more frequently found than the latter type in the DC area adjacent to the sensory epithelium and in the mid portion of the DC area. However, the latter type of DC was more predominant in the DC area distant from the sensory epithelium. These findings suggest that the activity of DCs is modulated by the melanocytes and that the former type of DC has a more active role in ion and/or fluid transport than the latter type of DC. Thus, the DC area near the sensory epithelium might be more actively engaged in the production and regulation of vestibular endolymph than that distant from the sensory epithelium. PMID- 7478451 TI - Effect of prolonged hypergravity on the vestibular system: a behavioural study. AB - Golden hamsters were exposed to conditions of 2.5 times normal gravity (hypergravity, HG) for 4 months. During this period, tests were carried out to study equilibrium maintenance, swimming behaviour and open-field behaviour of these HG hamsters and of control hamsters living in a normal-gravity environment. The tests proved to be useful devices for detecting differences in perceptive motor behaviour between HG hamsters and control hamsters. The HG hamsters had more difficulties in balancing on tubes and orientation during swimming. In the open-field study, the HG hamsters showed less locomotor activity than control hamsters. However, no differences were observed between the groups in washing, rearing and number of times having defaecation. These findings indicate that the daily transition from 2.5 to 1 g was not experienced as stressful by the hamsters, although performance on several perceptive-motor tasks was decreased, especially during the first weeks. PMID- 7478452 TI - The regeneration of the middle ear mucosa of the rat. AB - The purpose of the study was to investigate the regeneration of middle ear mucosa following surgical removal and to observe the effects of Silastic sheetings on regeneration. Two weeks after the operation the middle ear cavity was covered by moderately differentiated epithelium in the animals which had only the mucosa removed. In animals with inserted Silastic sheeting, infection slowed down the regeneration process. During the 2 months of postoperative observation, the surface of the cavity appeared to be rough, and there was intensive fibrin deposition over the mucosa. It is concluded that normal middle ear mucosa regenerates rapidly after surgical removal. On the other hand and in contrast to the purpose of its usage, Silastic sheeting slows down the process and causes fibrosis. PMID- 7478453 TI - Modified canal wall-up mastoidectomy with mastoid obliteration for severe chronic otitis media. AB - Mastoid tympanoplasty with a canal wall-up technique provides better conditions for a healthy skin lining than canal wall-down techniques. The formation of retraction pockets in residual pneumatized cavities is however a major cause of cholesteatoma recurrence with canal wall-up techniques. We have therefore attempted to combine the advantages of both canal wall-down and canal wall-up techniques. The posterior wall is kept up, the mastoid is obliterated with adipose tissue, the attic is exteriorized in the ear canal and hearing is restored with a minimal-size hypotympanic cavity only. Preliminary results from 41 cases with a follow-up of 6 months to 6 years suggest that skin conditions in the enlarged ear canal are adequate and that cholesteatoma do not recur. PMID- 7478454 TI - Borreliosis as a cause of peripheral facial palsy: a multi-center study. AB - Borreliosis is known to be a common cause of peripheral facial palsy in Stockholm and its vicinity. The aim of the present study was to investigate the frequency and characteristics of borreliosis among patients with peripheral facial palsy in different parts of Sweden. All serological tests were performed in one laboratory. Ten Swedish Ear Nose and Throat clinics participated in a prospective 1-year study of patients seeking medical attention for acute peripheral facial palsy. Twenty-eight (6%) out of totally 446 patients fulfilled the criteria for the diagnosis of borreliosis. The frequency varied between 1 and 16% and was highest along the southeast coast of Sweden whereas no case was reported from the northern part of the country. Borreliosis was more common among children with facial palsy than among adults. The infection occurred during all seasons although it appears to be less frequent during the spring months. Only a minority of the borrelial patients had a history of a preceding tick bite or erythema migrans. The fairly low overall frequency of this secondary stage of borreliosis in the study may be a result of better knowledge of the disease and earlier treatment of its early manifestations. In Sweden's endemic areas borreliosis is a common cause of peripheral facial palsy, and therefore all patients with facial palsy in these regions should be examined for borrelial infection. PMID- 7478455 TI - Effect of local tranexamic acid gel in the treatment of epistaxis. AB - In patients with nosebleedings, the hemostatic effect of local application of tranexamic acid gel or placebo was compared in a randomized, double-blind, multicenter clinical trial with parallel groups. The times needed to arrest the initial bleeding were recorded, as well as any rebleedings within 10 days. The results showed no significant differences in any of the efficacy variables. Tranexamic acid was no better than placebo in the early treatment of nose bleedings, but the gel itself seemed to have a beneficial effect. The gel preparations were easy to insert into the nasal cavity and caused no discomfort to the patients. PMID- 7478456 TI - Comparison of multiple-antigen simultaneous test and CAP systems for diagnosis of nasal allergy. AB - The multiple-antigen simultaneous test (MAST) is a simple system that uses no radioactive agents and allows simultaneous examination of multiple antigens. CAP is a quick new, in vitro system that is more sensitive than the radio allergosorbent test (RAST). To evaluate their clinical efficacies, we examined the correlation between the MAST and CAP systems. Serum samples were collected from 33 patients with nasal allergies, 13 males and 20 females, mean age 31.1 years. The MAST and CAP were used for 7 inhaled allergens: house dust, Dermatophagoides farinae, Japanese cedar, timothy, sweet vernal grass, ragweed and mugwort. The correlation coefficients found for MAST and CAP were significant for all the allergens tested. In addition, high values for sensitivity, specificity and efficiency were obtained for all the allergens. The MAST system provided the same information as the CAP system. Although CAP tended to have better sensitivity, some of its positive results may clinically be false positive. We believe that the MAST and CAP are both useful for the detection of allergens but that the diagnosis of allergy must be based on results of detailed examinations such as use of the skin test, the nasal provocation test and clinical symptoms. PMID- 7478458 TI - The normal wall of the esophagus as seen by endoluminal ultrasonography. AB - Endoluminal ultrasonography of the esophagus offers a possibility to study the esophageal wall, even in pathological disorders with narrow lumina such as benign or malignant strictures, by using 2.7-mm-wide probes. In order to evaluate the normal variations in size of the esophageal wall components, 10 volunteers and 10 deceased humans without upper gastrointestinal disease were studied with a 2.7-mm wide endoluminal ultrasonic probe. A comparison with a histopathological technique was also performed. The results indicated a good correlation between the volunteers and the deceased in using endoluminal ultrasonography. In contrast a discrepancy was noted between the measurements made by ultrasonography and histopathology. PMID- 7478457 TI - Effects of adenotonsillar hypertrophy on snoring in children. AB - A prospective study was performed to evaluate the effects of adenotonsillar hypertrophy on snoring in children. Thirty male patients were grouped with respect to the severity of snoring and were evaluated in terms of the noise level of the snoring sound, the lowest arterial oxygen saturation, degree of palatine tonsillar hypertrophy, body mass index and cephalometric findings that included the adenoidal-nasopharyngeal ratio, the angle between the lines sella turcica/nasion and most posterior point of anterior maxilla/nasion, the angle between the lines sella turcica/nasion and most posterior point of anterior mandible/nasion, the posterior airway space, the distance from the sella to the nasion, lower face height and the distance from the basion to the posterior nasal spine. The noise level of the snoring sound, the lowest arterial oxygen saturation and the adenoidal-nasopharyngeal ratio showed a significant correlation with the severity of snoring, but the degree of palatine tonsillar hypertrophy and the body mass index failed to disclose any significant relationship. PMID- 7478459 TI - Arteriovenous malformation with auricular hypertrophy. AB - We report a case of arteriovenous malformation (AVM) restricted to the auricle fed by the external carotid artery. The patient showed severe hypertrophy of the left auricle, remarkably spreading throughout the auricle. At first, we chose to surgically ligate the superficial temporal artery. After the operation, however, pulsation recurred, and we ligated the external carotid artery at an inframandibular site in a second operation. Two months after surgery, plasty of the left auricle was performed at the patient's request. PMID- 7478460 TI - The common cavity congenital deformity of the inner ear. An important precursor of meningitis described in 1838. AB - Congenital deformity of the osseous labyrinth of the inner ear is well recognized as being associated with a fistulous communication with the intracranial subarachnoid space in some cases, leading to meningitis or a spontaneous 'stapes gusher'. Such cases have been described inappropriately as 'Mondini's dysplasia'. We cite 2 patients who both died of meningitis and had an accurate postmortem anatomical appraisal of the deformed temporal bones, one recently described by us and one by Cock more than 150 years ago. The use of eponyms is to be deprecated but should faithfully reflect the original description. The 'common cavity' lesion of the inner ear is a most important precursor of otogenic meningitis readily assessed by computed tomography and needing surgery to prevent the egress of cerebrospinal fluid or ingress of pathogens causing meningitis. PMID- 7478461 TI - Twin pregnancy following in vitro fertilisation coinciding with laryngeal cancer. AB - The authors report the case of a 33-year-old female patient who was admitted to their Department with supraglottic laryngeal cancer detected in the 1st trimester of her twin pregnancy following in vitro fertilisation after a 10-year nulliparous period of her married life. Since a horizontal supraglottic laryngectomy performed during pregnancy the patient has been free of recurrences, and the twins delivered 3 years ago are in good health. According to the authors, of all available therapeutic possibilities (surgery, cytostatic drugs, telecobalt irradiation) in pregnancy coinciding with laryngeal cancer, optimal therapeutic effects can be achieved by surgery. PMID- 7478462 TI - [Most frequent causes for hand tremor in clinical practice]. AB - The author summarises the various origins of hand tremor. Physiologic tremor can be demonstrated in any individual. This tremor is normally recordable at a stable frequency of 8-10 Hz, though this can be influenced by age, the amplitude can vary substantially. Physiological tremor is believed to have its origin in the mechanical properties of the extremities, and the firing properties of motoneurones. Essential tremor is an inherited autosomally dominant phenomenon, and is characterised by action and postural tremor. It is progressive with age. The use of low doses of theophylline in the treatment of essential tremor is emphasised, in view of the low incidence of side effects. A hypothesis is also presented to account for essential tremor in terms of an imbalance in the activity of excitatory and inhibitory amino acid system. Parkinsonian tremor may begin as postural or resting tremor on one side, and it is reduced by active movement of the affected extremity. The prognosis of Parkinson's disease presenting with tremor is better when presenting with hypokinesia. The increasing use of non-specific drugs in the treatment of Parkinsonism, and the use of thalamic high frequency stimulation are indicated. PMID- 7478463 TI - [Changes in the frequency of childhood brain tumors in North-East Hungary]. AB - Authors report about a seemingly increase of the number of pediatric brain tumors in Borsod county region North-East Hungary. In a retrospective study they conclude, that this phenomenon may to a great extent be artificial, that is, it may reflect the improvement of diagnostic possibilities, above all the new imaging modalities. At their Institution the pattern of brain tumors is similar to that reported in the literature. In addition a racial difference has been detected, since the gipsy population of this region seems to have lower incidence of pediatric brain tumor than the Hungarian majority. PMID- 7478464 TI - [Monitoring somatosensory evoked potentials during carotid endarterectomy]. AB - The tromboembolic and ischemic events during carotid endarterectomy can be avoided or detected with appropriate monitoring. Median nerve somatosensory evoked potentials recorded from the parietal cortex correlate with the blood flow in the middle cerebral artery. The good evoked responses after cross-clamping of the carotid artery indicate a sufficient collateral circulation, enabling a surgery without shunt, thus minimizing the risk of embolisation. Insufficient collateral circulation after cross-camping results in an amplitude reduction of the parietal N20-P25 complex of more than 50%. In this case an ischemic event can be prevented by shunting. As a total 83 carotid endarterectomies were done. In 65 cases (78.3%) the evoked potentials showed no significant alteration, and no postoperative neurological deficit occurred. Seven patients (8.45%) needed to be operated with a shunt, because of cross-clamping ischaemia. One of them presented a transient postoperative hemiparesis, which was predicted by the long-term loss of the SEP-s, and which resolved within 4 hours. Seven further patients (8.45%)- operated primarily with shunt, and 4 patients (4.8%)--monitored with transcranial Doppler sonography, showed no postoperative neurological deficit. We found that median nerve somatosensory evoked potential monitoring during carotid endarterectomy is a simple, sensitive and reliable method. PMID- 7478465 TI - [Detection of Helicobacter pylori in biopsy specimens, methodical studies]. AB - Demonstration of Helicobacter pylori infection receives more and more importance in nowadays gastroenterological practice. The authors have compared culture and histology from 69 antral biopsy specimens for their ability to document Helicobacter pylori infection. Infection ratios in the context of clinical and histological diagnoses resulted in a distribution pattern similar to that described by others: 85-69% of duodenal ulcer patients, 67-67% of gastric ulcer patients, 62-54% of patients with gastritis and/or erosion(s) and 33-60% of endoscopically negative patients were found to be Helicobacter pylori positive with culture and histology respectively. Normal or atrophic mucosa showed no bacteria with either methods, but one must also consider the small number of such cases in this study. Chronic gastritis with no signs of activity proved to be infected only in a minority of cases, while chronic active gastritis cases were Helicobacter pylori positive in 72 and 61% histologically and with culture respectively. The modified Giemsa stain used in this study grave a relative specificity of 0.74 and sensitivity of 0.79 as compared with culture. There was a good association between the two methods tested, and this could be further improved by doing the two tests simultaneously. The association was weaker when the tests were done asynchronously, this is why it is not recommended to use such a diagnostic schedule. Comparison of the traditional haematoxylin-eosin stain with the modified Giemsa stain resulted in a very strong association between the two.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7478466 TI - [Bronchiolitis obliterans organizing pneumonia: survey based on two cases]. AB - Authors offer a survey about an aspect forming a clinical entity called bronchiolitis obliterans organizing pneumonia, by two of their patients' presentation. In Case 1 the process manifested in form of a disseminated, small spotted, diffuse shadow, causing serious resting hypoxia. Diagnosis was supported by histological examination of a substance obtained by small surgical thoracotomy. Five months' therapy of Imuran and altogether 15 months' therapy of Prednisolon resulted in recovery. Second patient's symptoms and the shadow representated in the left upper lung field on chest X-ray, proposed the probability of a tumour. The lesion removed surgically, together with the left upper lobe, proved to be the clinical appearance that constitutes histologically the subject of this paper. Surgery alone resulted in definitive recovery. Disease can be proved on the basis of histological picture and it's important to sunder it primarily from bronchiolitis obliterans as well as from other interstitial pulmonary diseases. PMID- 7478467 TI - [Obstetrical anesthesia in Hungary]. PMID- 7478468 TI - [Participation in the European Union Program for the Prevention of Colonic Polyposis]. PMID- 7478469 TI - [In situ hybridization for the detection of hepatitis C virus RNA in mononuclear cells of peripheral blood from infected patients]. AB - In situ hybridization was used to investigate hepatitis C virus infection in peripheral blood mononuclear cells in 11 patients with chronic hepatitis. Using 35S labeled HCV-RNA probe, HCV-RNA positive and negative strands were observed in unstimulated cells of 3/11 patients; all 3 being treated with immunosuppressive drugs after orthotopic liver transplantation. HCV-RNA sequences were also identified in mononuclear cells obtained from 3 patients without immunosuppression, after stimulation of the cells with either phytohemagglutinin or pokeweed mitogen. In contrast, HCV-RNA was not found in 5 cases, where there was no liver transplantation or cell stimulation by mitogens. These results definitely assess infection of mononuclear cells by HCV. In addition, they demonstrate that mitogenic stimulation of infected cells increases HCV-RNA replication. PMID- 7478470 TI - [Cilazapril replacing captopril in the therapy of medium-severe essential hypertension]. AB - Authors performed an open, crossover, multicenter study of oral cilazapril versus previous captopril treatment in mild to moderate hypertension. The treatment of the 100 outpatients on daily three or four times captopril was found ineffective, or in some cases side effects or non compliance necessitated a switch to a once daily dose of cilazapril. Reasons of ineffectivity were compliance problems in 76% of the patients during long term captopril therapy. Blood pressure decreased from 163.28 +/- 14.5/97.5 +/- 9.35 mmHg on captopril therapy to 136.67 +/- 12/83.49 +/- 7.77 mmHg at the end of a 12 week cilazapril treatment (p < 0.001). 80 patients received cilazapril monotherapy (with doses of 2.5 mg in 54 cases, 5 mg in 18 patients). 7.5 mg in 4 cases, 1 and 1.25 mg in 2 patients. In 20 patients an adjunctive diuretic was also added, while the cilazapril treatment was ineffective in 3 patients. In respect of the global evaluation and scoring of cilazapril versus captopril therapy, a clear and statistically significant improvement could be demonstrated in efficacy, tolerability and compliance after a 12 week cilazapril treatment. The 24 hour ambulatory blood pressure measurement performed in 13 patients also verified a decrease in blood pressure achieved by cilazapril therapy. Authors conclude that in case of ineffectivity of three or four times daily captopril treatment (caused most likely by non-compliance), a switch to a once daily dose drug like cilazapril is indicated. PMID- 7478471 TI - [Characteristics of HLA II antigen expression in the rectal mucosa of children suffering form ulcerative colitis and Crohn disease]. AB - The staining of HLA class II. antigens was studied with monoclonal antibodies using sensitive immunohistochemical methods in the rectal biopsy specimens taken from 4 children with Crohn's disease and 9 with ulcerative colitis as well as from 10 controls. HLA class II. antigen expression was not seen in the surface and crypt epithelium of controls. The majority of patients showed epithelial HLA DR, and HLA-DP expression, fewer had HLA-DQ expression. In patients who had more severe clinical and histological symptoms the epithelial HLA-DR and HLA-DP expression was more intense, than in those with milder presentation (p < 0.05). We conclude that the activity of ulcerative colitis may be infered from the intensity of HLA class II. antigen expression. PMID- 7478472 TI - [The role of outpatient service for postmenopausal women and its use in the screening for hypertension and its risk factors]. AB - The authors studied the risk factors of hypertension in women treated at the outpatient postmenopausal care unit in the town Csongrad. Anamnestic data, serum parameters were compared in the hypertension and in the normotension groups. 40% of women with hypertension and 35% of women with hypercholesterinaemia have been discovered for the first time at the postmenopausal care unit. The authors emphasize the important role of the postmenopausal care unit in screening of some risk factors of hypertension--overweight, smoking, hypercholesterinaemia--and in screening of the hypertension itself. PMID- 7478473 TI - [Neurofibromatosis (von Recklinghausen disease) and carcinoid tumor in Vater's ampulla]. AB - The localization of the carcinoid tumors of the gastrointestinal tract--as a part of the APUD system--is extremely seldom at the region of ampulla of Vater. A case of congenital neurofibromatosis is reported in a 49-year-old male patient, with a carcinoid tumor of the ampulla of Vater in the background of long-standing, atypical biliary complaints. Periampullary neoplasm of neural-crest origin are very rare, up to this time 73 cases of Vater papilla carcinoid tumors were described. Regarding the development of VP carcinoid in neurofibromatosis, our patient is the 21st reported case in the world literature, and the first one in Hungary. The authors review the literature, giving special attention to the risk of periampullary neoplasms in von Recklinghausen's disease, the need of accurate diagnosis and the correctly selected operative intervention. PMID- 7478474 TI - [Hermann von Helmholtz: inventor of the ophthalmoscope]. PMID- 7478475 TI - [Dezso Miskolczy, his work in neuropathology]. PMID- 7478476 TI - [Second malignant disease in patients under treatment for malignant lymphoma]. AB - Between 1978 and 1993, 1013 patients 529 with Hodgkin's disease and 484 with non Hodgkin's lymphomas--were treated in our department. Out of 1013 patients secondary neoplasms developed in 23 cases: 3 acute nonlymphocytic leukemia, 19 solid tumors and 1 secondary non-Hodgkin's lymphoma. The median time from diagnosis of malignant lymphoma was 7 years and their median age was 49 years. 11 patients with secondary tumor were treated with chemotherapy and 12 received combined (radio- and chemo-) therapy. Since alkylating agents increase the risk of leukemia and radiation contributes mainly to other cancer, future treatment protocols should attempt to reduce the most serious consequences of therapy without compromising the survival. Careful lifelong observation is indicated for patients with malignant lymphomas, with special attention given to new clinical signs or symptoms. PMID- 7478477 TI - [Enhanced accuracy of the X-ray diagnosis of acute abdomen with patient in the supine (pregnancy standard) position]. AB - The value of exposed to vertical central ray supine (dorsal decubitus or pregnancy standard) plain films was determined in this retrospective study in diagnosing pneumoperitoneum based on comparing of exposed to horizontal X-ray beam abdominal radiographs in 54 patients associated with perforation of hollow viscus (48 stomachs or duodenums of them). Until the anteroposterior erect or sitting and semi-recumbent positioning views moreover laterolateal supine and posteroanterior left lateral projections were negative in all cases, then in 24 of 54 subjects free intraperitoneal gas collections as spotted radiolucencies were detected inside the right upper quadrant of abdomen on pregnancy standard dorsal aspects. The orientation of gas to left upper abdominal quadrant was noted in additional 2 cases. The 18 of 26 perforations were not proceeded to free intraperitoneal space. The pregnancy standard plain film is a sensitive technique for visualisation of pneumoperitoneum after perforation of the hollow viscus between 3-6th penetration hours just as in unable to cooperation patients, particularly. PMID- 7478478 TI - [Prenatal diagnosis of left cardiac abnormality]. AB - We found out of 1500 prenatal cardiac ultrasound examinations very small left heart in 14 cases. Six of the 14 cases were hypoplastic left heart syndrome and eight functional-secondary left heart hypoplasia. The most important signs the echocardiography in cases of hypoplastic left heart are as follow: hypoplasia of mitral valve with or without measurable flow, absence of the anterograde flow through the aortic valve, retrograde flow in the isthmus. In cases of functional left heart hypoplasia the size of the left ventricle, aorta and mitral valves were under the 3rd percentile but there were measurable anterograde flow through the aorta. In eleven cases there were other malformations: hygroma colli, diaphragmatic hernia and omphalokele. The differentiation of the hypoplastic left heart from the secondary left heart hypoplasia has a great importance because of the therapic planning and prognosis. PMID- 7478479 TI - [Questions about re-sterilization and reuse of sterile disposable devices]. AB - Authors deal with the problems of restilization and reuse of sterile disposable devices on the basis of literary data, their own experiences, and the standpoints offered on the Fourth Symposium of the Canadian Office for Health Technology Assessment. They state, though the reuse of sterile disposable items is possible to work out, it needs a very sophisticated technology, its thriftiness is often doubtful, and the related legal and ethical problems are mostly unsettled. The personal, material and legal preconditions assuring a safe reuse do not exist in Hungary at present. PMID- 7478480 TI - [Unstable angina pectoris associated with surgically treated coronary artery atresia]. AB - A case report of a rare, presumably congenital form of "single coronary artery", is presented. The authors review the classification and clinical significance of the entity with special emphasis with regard to anginal symptoms and the so called sudden unexpected death syndrome. Details and result of surgical intervention (conventional aorto-coronary saphenous bypass) are discussed. PMID- 7478481 TI - [The role of physical activity in the prevention of certain internal diseases]. AB - During the last decades the physical activity drastically was reduced in daily life. Epidemiological and prospective studies suggest that regular physical activity has beneficial effects in prevention of non-insulin dependent diabetes mellitus, and has protective benefit in control of risk factors in coronary artery disease. It increases cardiovascular functional capacity, decreases myocardial oxygen demand and has a blood-pressure lowering effect. In the prevention of obesity together with a proper diet, regular training is the most important factor influencing life-style. Some studies indicate that exercise might promote protection against colon, breast and lung cancers. To maintain good health aerobic exercise with a moderate intensity on a regular basis is recommended. PMID- 7478482 TI - [Cases of diseases registered with the Genetic Counseling Service during the past 20 years, in the light of their etiology]. AB - The authors analyse the experiences of their Genetic Counseling Clinic between 1979-1992. They show the formation of the motives of presenting and consulting. They range the consultants in three etiological categories (genetical, teratological and clinical entities). The proportion of new consultants increased fourfold and this tendency documents unambiguously the claim of family planners for this very important medical service. The proportion of Mendelian disorders, mutagen risk and consanguinity are relatively continuous. Because of teratogen risk the rate of presenting decreased moderately. The proportion of consultants shows a definite increase of multifactorial frequent disorders, which they value as a favorable tendency. For decreasing of unjustified cases they suggest the necessity of pre-screening. PMID- 7478483 TI - [Comparative study of superoxide production of monocytes in primary biliary cirrhosis]. AB - Monocytes appear to play a role in immunological abnormalities observed in primary biliary cirrhosis (PBC). Monocytes not only produce fibroproliferative factors, such as IL-1, TNF, and PDGF but also produce superoxide anion which can directly damage tissues, and thus may lead to fibrosis. The aim of this study was to compare the superoxide production in monocytes obtained from 12 control persons, 9 patients with non biliary cirrhosis, 6 untreated PBC patients, 6 patients with gallstones under urso- and chenodeoxycholicacid (Lithofalk) treatment and 32 PBC patients under ursodeoxycholicacid (UDCA) therapy. Monocytes were isolated and the production of superoxide anions with and without phorbol myristate-acetate (PMA) stimulation was determined. In two occasion, the monocytes from control patients were preincubated with 10, 50, 100 microM UDCA. Unstimulated monocytes from PBC patients under UDCA therapy produce an average 43% more and the PMA stimulated monocytes an average 42% more superoxide than monocytes from the control or from the other cirrhotic patients. The UDCA preincubation did not influence the superoxide production of monocytes obtained from control patients. These findings suggest that the increased activity of monocytes may also play a role in liver damage and fibrosis in PBC. PMID- 7478484 TI - [Cranioplasty using computer-designed implants (preliminary report)]. AB - The co-operation among plastic surgeons and computer experts is presented demonstrating the reconstruction of a cranial bone defect of a patient. The adequacy of manufactured implant for reconstruction of cranial bone defect on the base of classic methods in only approximate. Computer aided design is needed for creating a more sophisticated implant. Cranial CT is digitised and a 3 dimensional model is created. The precise implant, manufactured by a computer directed milling machine, fits ino the bone defect. During the operation the cranial bone defect is explored, the bone edges are identified. The implant is fixed and the skull skin is closed. The authors demonstrate a successful operation of a cranial bone defect of 9 x 6.7 cm, the complicated shape and bone surface was reconstructed by a computer aided and manufactured implant. The co operation of plastic surgeon and computer experts are essential during the process of implant design. The computer assisted and manufactured implants are suitable for the most sophisticated functional and aesthetic reconstruction of bone defect with complicated surface and irregular pattern. PMID- 7478485 TI - [Significance of the length of the fetal femur]. AB - The importance of the examination of fetal skeletal system, especially the measurement of femur length must be emphasized during the routine ultrasound screening. A case of osteogenesis imperfecta is described, diagnosed on the 18th week of pregnancy by ultrasound. PMID- 7478486 TI - [Erno Jendrassik, internist-neurologist, his interest and activities in the field of psychiatry]. PMID- 7478488 TI - [Chemonucleolysis: taking a position. Expert discussion, 30-31 October 1992]. PMID- 7478487 TI - [J. de Carro (1770-1857): Eurasian prophet of Edward Jenner]. PMID- 7478489 TI - [Knee endoprostheses--axial knee or surface replacement? Workshop, Paris, 22 April 1993]. PMID- 7478490 TI - [Role of antibiotics in the treatment of infected joint prosthesis]. AB - Infection is a rare, but extremely severe, complication of prosthetic joint surgery. Until recently, antimicrobial agents were not generally used in the management of such infections. Antibiotics now have an important role, either combined with replacement surgery or even as the only treatment in selected cases. In earlier studies, high failure rates were reported with conservative therapy. These unsatisfactory results were probably due to a lack of collaboration between surgeons, infectious disease specialists and microbiologists. All patients with a long history of infection or with loosened implants should undergo joint replacement. Early or rapidly diagnosed hematogenous infection in patients with stable prostheses can be treated conservatively. In most cases, such a treatment is preceded by revision surgery, which is needed for microbiological diagnosis and for debridement. The choice of antibiotics depends on the microorganism involved and the results of susceptibility testing. The most important etiologic agents are Staphylococcus aureus and coagulase-negative staphylococci. Antimicrobial drugs used in device related infections should act on surface-adherent and stationary-phase bacteria. In an animal model, rifampin combined with a quinolone has proved to have the highest cure rate against staphylococcal foreign-body infection. Rifampin is indeed highly efficacious on surface-adherent and stationary-phase bacteria. These experimental data were confirmed in clinical studies; cure rates of 60-80% were observed with rifampin combinations without joint replacement. Antimicrobial therapy should be continued over at least 3 months in hip implant infection and at least 6 months in knee implant infection. Before treatment is stopped, signs and symptoms of infection must have been absent with C-reactive protein normal for at least 1 month. PMID- 7478491 TI - [Infected hip joint prosthesis. Viewpoints for 1-stage and 2-stage prosthesis exchange]. AB - When treating an infected hip prosthesis, there are a number of medical, technical and biomechanical considerations. No ideal solution for all problems is available. In individual cases an optimal compromise must be found. The means for curing the infection and the one for maintaining hip stability can counteract each other. Basically, three options are available: removal of the prosthesis without replacement and one- or two-stage replacement. With all three options thorough surgical debridement is vitally important. The introduction of antibiotic cement and cementless titanium implants, together with very effective local antiseptics, has remarkably improved our treatment of this problem. Therefore, removal of infected prostheses without replacement is not as well accepted now as it has been in the past. PMID- 7478492 TI - [Local antisepsis in infected total hip endoprosthesis]. AB - Between 1991 and 1993, 34 infected total hip prostheses were treated with the topical antiseptic hexamethylenbiguanide (Lavasept) in addition to thorough debridement of infected and necrotic tissue and systemic antibiotic administration. In 4 cases (group 1) a contaminated hematoma was treated within the first 3 weeks after joint replacement. Soft tissue revision without removal of the prosthesis was performed in 6 cases (group 2). In 18 cases (group 3) a one stage exchange of the infected prosthesis was done. In 6 cases (group 4), local and general conditions were so poor that the prosthesis was removed without replacement. The success rate was 100% in group 1, 20% in group 2, and 77% in group 3. In group 4, 1 patient died of respiratory problems 12 days after surgery while in 2 patients the infection healed uneventfully and in 1 patient healing was achieved after a second revision. An 81-year-old patient in poor general condition had an impressive improvement of his severe infection, although very minor drainage continued. Hexamethylenbiguanide has proven to be a useful supplement to thorough surgical revision. However, longer follow-up data are required. PMID- 7478493 TI - [Treatment of infected total prosthesis arthroplasty of the hip joint]. AB - With a median follow-up of 8 (1-13) years in our series, we are familiar with the entire courses in 62 infected hip arthroplasties, treatment starting between 1980 and 1986. The primary treatment was debridement with suction/irrigation in 6 cases, one-stage replacement of the prosthesis in 32 occasions and two-stage replacement in 23 cases. Only one definitive Girdlestone procedure was done. With further operations successful healing was finally achieved in all 62 hips. In 74% healing was primary, this being the case in 91% (10/11) of early and 71% (36/51) of late infections. Device-retaining procedures can be tried, particularly in early infections with low-virulence bacteria and a stable implant. One-stage revisions can be done in early and late low-virulence infections with loosening of components if bone and soft tissue are in good condition. In all other situations a two-stage procedure is recommended. As a rule, reimplantation can be done within 2 weeks. In cases with recurrent infection, replacement should be delayed for several months. As an alternative, several open debridements with reimplantation of a new prosthesis after 2 weeks can be considered. Girdlestone resection arthroplasty should be avoided whenever possible. Antimicrobial treatment is of great importance: it should be maintained for at least 3 months and in any case until 1 month after normalization of laboratory and clinical signs of infection. PMID- 7478494 TI - [Treatment of periprosthetic infection of the hip using one-stage exchange surgery]. AB - This article presents the theoretical microbiological principles involved in the pathogenesis of periprosthetic infection, the role of microbiology in diagnosing this disease, and factors requiring special consideration when selecting topical antibiotics. It also describes the operative technique of one-stage exchange arthroplasty using antibiotic-loaded acrylic cement and the results achieved with this method to date. PMID- 7478495 TI - [Total hip endoprosthesis following resection arthroplasty]. AB - From 1976 to December 1994, a total of 347 patients underwent implantation of a hip prosthesis at the ENDO-Klinik for treatment of an unsatisfactory condition following resection arthroplasty. From 1976 to 1987, 143 patients were treated and in 1989 the results obtained in these patients were analysed: 99 of them were available for a follow-up examination in 1989, and 64 for a further examination in 1995. In 130 cases infection had been the reason for joint resection. At the time of the prosthesis operation (1-20 years later) intraoperative biopsy revealed that infection was still present in 41 cases (31.5%). Only 15 of these infections had been detected preoperatively by joint aspiration. This shows that the value of resection arthroplasty as a method of treating periprosthetic infection is limited and lends support to the one-stage exchange operation, which is the method we prefer in cases of infected hip prostheses. The operative technique and preparation for implantation of the prosthesis are described, as are septic and aseptic complications and the measures that can be taken to treat them. In spite of the patients' generally poor initial condition and with due consideration for the further revision operations, the medium-term results finally obtained are poor in only 9%. PMID- 7478496 TI - [Salvage surgery in chronic infection following total hip prosthesis]. AB - Thirteen patients were analyzed who suffered from chronic infection after total hip replacement. Some of them had had severe soft tissue defects or multiple fistulae for years. Some showed acute, sometimes life-threatening infection after long-standing loosening of the prosthesis. Therapy consisted of radical debridement, installation of a closed suction-irrigation drainage system and if another total hip prosthesis was implanted of a specific antibiotic management, including rifampin. In one case treatment was limited to the achievement of a permanent fistula. The average follow-up time was 4.3 years. With or without total hip reimplantation infections were healed as a rule and the decrease in pain was marked. There was only a partial increase in walking ability. In these severely damaged patients treatment can result in marked amelioration of the quality of life but only exceptionally in normalization. PMID- 7478497 TI - [2-stage exchange in the treatment of infected knee prosthesis]. AB - We report our experience with the treatment of infected total knee arthroplasties with a two-stage procedure during the period between 1992 and 1995. The two-stage procedure as a cure for infected total knee arthroplasty is a reliable and reproducible method of treatment. The infection healed in 86% of cases, and all patients with a follow-up of more than 1 year were satisfied with the result achieved. Also the objective values proved that the outcome of this severe complication of total knee arthroplasty was favorable. The two-stage exchange procedure is recommended as the treatment of choice for infected total knee arthroplasty wherever possible. PMID- 7478498 TI - [Infection following shoulder and elbow arthroplasty. Diagnosis and therapy]. AB - The rate of infection reported in recent publications is 0.8% after shoulder arthroplasty and ten times higher (8.1%) after elbow arthroplasty. The figures for shoulder arthroplasty correspond well with our own rate of revision for infection of 0.5% after 363 primary shoulder replacements. However, our average rate of revision for infection (1.8%) after 278 GSB-III elbow arthroplasties was considerably lower and included rheumatoid as well as post-traumatic indications. Our experience concerning etiologic factors, nature, diagnosis, treatment options, and long-term consequences of superficial and deep infections after shoulder and elbow arthroplasty are discussed for each joint separately and in relation to the literature. PMID- 7478499 TI - [Julis Wolff and his "law of bone remodeling"]. PMID- 7478500 TI - [Basics of Pauwels' theory of the functional adaptation of bones]. AB - Bone is a secondary support tissue. It develops on the base of a preexistent primary scaffold, built up by connective tissue or by cartilage. Bone formation depends on the magnitude of the elastic deformation, due to loading. The skeletal organ "bone" reacts to the actual stress like a feedback system: high stress stimulates new bone formation and incorporation of mineral salts; low stress leads to bone resorption and mobilization of mineral salts. The result of extreme stress is bone resorption (stress fractures, pseudocysts). The radiological density and architecture of the spongy bone reflect exactly the direction and local magnitude of stress. PMID- 7478501 TI - [The significance of blood vessels on both sides of the growth plate]. AB - A new 3-D presentation of the whole vasculature of the epiphyseal growth plate clearly allows for a distinction between arterial and venous sections of the blood circulation. Histological investigations using these processing methods could neither prove the connection between epiphyseal and metaphyseal vessels through the growth plate in newborns nor the concept of an arterial epiphyseal blood supply of the growth plate presented by Trueta. It is rather a question of an arterial metaphyseal perfusion and an epiphyseal venous drainage system. PMID- 7478502 TI - [Biomechanical reaction of the bones to intra- and extramedullary load bearers: the significance of implant contact]. AB - Bone reacts very differently to changes in the environment relating to implant surgery. Since the advantages and disadvantages of implants have to be set against each other, the functions of implants and the demands made upon them will be considered first. The contact area between the bone and the implant will then be discussed. The contact surfaces and their induction of necrosis are at present in the forefront of our research in the area of biomechanics. The essential issues are: the avoidance of early temporary porosity, the possibilities of providing best conditions for "accelerated" bone healing, the reduction of refracture risks, and the improvement of infection resistance. Considerable opportunities for improvement of the most recent insights into the biology and biomechanics of the contact zones. PMID- 7478503 TI - [Endoprosthesis and bone reaction]. AB - Prosthetic implants cause a reaction on the part of the surrounding bone. Bone reaction can be interpreted as a dynamic response of the internal bone architecture to stress and strain. On the other hand, instability at the bone implant interface or biological reactions to wear particles may lead to bone resorption. Standard X-ray examination has remained the most important tool for evaluation of the bone-implant relationship. Knowledge of the radiological features of bone remodelling and bone reaction is therefore a prerequisite for the orthopaedic surgeon. Based on eight case reports after prosthetic replacement of the hip, important radiological signs of bone reaction are presented. If possible a biomechanical or biological explanation is attempted. The information drawn from these cases, as well as from the current literature, suggests that after prosthetic joint replacement routine X-ray follow-up is required, even in asymptomatic patients, and that progressive osteolysis requires early operative revision, even if there are no acute clinical problems. PMID- 7478504 TI - [Stress and fracture healing]. AB - Stress is an important factor in fracture healing, but it is not the only one. The exact amount of stress, displacement or strain needed or tolerated in fracture healing is not yet clear. There are numerous modifying factors influencing fracture healing like fracture type, soft tissue injury, vitality of fragments, type of osteosynthesis, soft tissue handling, localization of injury, activity level and weight of the patient, local geometry and muscular topography. However, despite our large deficits in knowledge of the process of fracture healing, typical patterns of failure or disturbed healing can be recognized. This could help bringing stress and fracture healing in balance and includes an analysis of pathological stress and our efforts to neutralize, canalize or use it to our advantage. PMID- 7478505 TI - [Traumatic loosening of apophyses in the pelvic area and the proximal femur]. AB - Avulsion fractures of the pelvis and the proximal femur are relatively rare injuries in the adolescent age group of 11 to 17 years. The overall prognosis for spontaneous healing is good. The injury most frequently involves the apophysis of the anterior inferior iliac spine and of the lesser trochanter, followed by anterior superior iliac spine. In our experience the avulsion fractures involving the ischial tuberosity and the greater trochanter are extremely rare. The latter can have dire consequences. PMID- 7478506 TI - [Callus distraction]. AB - Fibroblast networks that form collagen and connect the two ends of bone develop in the haematoma after corticotomy. This regenerative tissue is vascularized and distracted. Even during the lengthening, mineralization starts. This starts at the ends created by the osteotomy, in the form of conical bony columns 200 microns thick, which grows towards each other in a manner reminiscent of stalagmites and stalactites, until the central, fibrous inner zone (growth zone) formed during distraction is completely mineralized. Connective tissue and bony bars are arranged lengthwise. As a rule, intramembranous callus formation takes place during distraction osteogenesis with no intermediate cartilaginous step. PMID- 7478507 TI - [Bone hypertrophy as a results of training]. AB - It is well known that adaptational processes following athletic training affect not only muscles and tendons, but also the skeletal system of the athlete. Functional adaptation of the bone is shown by an increase in cortical thickness, frequent development of bony spurs at the insertion of tendons and joint capsules, an increase of density of the bone substance, and bony changes as a reaction to increased mobility of bony junctions such as the symphysis of the pubic bones of the sacro-iliac joints. Recently, in an investigation involving 20 professional tennis players, we demonstrated that--in addition to the adaptational processes mentioned above--stimulation of longitudinal bone growth and widening of the joint space (cartilage hypertrophy) occur. The possibility of bony hypertrophy as a result of athletic training, especially with regard to increases in of bone density and bone diameter, should lead to incorporation of sports medical considerations into the prophylaxis and therapy of osteoporosis. PMID- 7478508 TI - [Lesions of the growth plate caused by sports stress]. AB - Besides the positive physiological, psychological and social aspects, sports activities in adolescents bear the risk of injuries and overuse of the locomotor system. Previous examinations have shown that increased stress to the growth plates by sports activities, in relation to the intensity of strain during growth spurts, can influence normal growth. In female gymnasts, hormonal changes can decrease the growth speed and long-term growth. On the other hand, during more intensive phases of growth the column cartilage of the growth plate is the weakest part of the locomotor system because of the influence of somatotropin and low levels of testosterone. This can cause subchondral stress fractures in the growing cartilage that later on, if missed or not sufficiently treated, can cause osteochondrosis dissecans. The apophysis of tendons of big muscle groups can show loosening of the apophysis caused by increased muscle strength and acute or chronic microtrauma. Male adolescents show an incidence of lesions in the relation of 9:1 to female adolescents. The therapy for apophyseal lesions is generally nonoperative. Due to the persistent growth possibility, pseudotumors can occur, which can cause problems in differential diagnosis among skeletal tumors. Too high pressure, pushing and tearing forces can influence growth. Later examinations of previous high-level sportsmen and patients with coxarthrosis with and without a sports history show that blockage of the rotation of the foot during growth, for example caused by soccer shoes, can cause high pushing forces on the femoral epiphysis, which can lead to epiphyseolysis cap. fem. lenta and thereby to pre-arthritic deformities. This is overcorrection of the "physiological" epiphyseolysis, described by Morscher. Knowledge of the reduced strength of the growth plate indicates better adaptation of training and supervision of the adolescent high-level sportsman. A regular check-up of the growing athlete and a reduction in sports intensity during the growth spurts, prohibition of negative training parts and sometimes even prohibition of sports at all, if there is a lesion of the growth plate or hormonal disorders, are sometimes necessary to minimize late defects. In addition to this, a reduction of strain in some sports and, for example, prohibition of rotation-blocking soccer shoes in the adolescent soccer player is necessary. PMID- 7478509 TI - [Stress fractures in athletes and military recruits. An overview]. AB - Stress fracture is an overuse injury caused by muscle forces together with bending and impact forces acting on the bone, which has not adapted to the loading. The localization of stress fractures is more common in lower extremities. They are found in many other bones of the body as well. In the history of these patients a considerable amount of running exercise is usually found. The symptoms are stress pain and aching at rest after training. Typical findings are local palpation pain and edema. Sometimes tender resistance is felt. Clinically used radiological imaging methods are radiographs, scintigraphy and in some cases magnetic resonance imaging and computed tomography. The radiographs are not usually positive until 2 weeks after the onset of symptoms. With a typical history and clinical findings the radiological diagnosis causes no further problems. The corner stones of the treatment are: early identification of the symptoms, early diagnosis, a sufficiently long training pause and in special cases consultation of experts in the field. There are rare stress fractures with increased risk of a delayed union, non-union or complete fracture. Surgical treatment may be needed in some cases. The prevention of stress fractures has proved to be difficult. The risk of acquiring stress fractures is increased by running with improper shoes and in female athletes with menstrual irregularities. If running is changed to some other kind of sport, the incidence stress fractures is lower. PMID- 7478510 TI - Direct interaction of Gadd45 with PCNA and evidence for competitive interaction of Gadd45 and p21Waf1/Cip1 with PCNA. AB - We have previously shown (Smith et al., 1994) that antibodies raised against the growth arrest and DNA damage inducible protein Gadd45 co-precipitate proliferating cell nuclear antigen (PCNA), a protein involved in DNA replication and repair. Here we demonstrate that Gadd45 can directly bind to PCNA using a Far western blotting approach. In this assay, a Gadd45 bacterial expression vector was modified to allow synthesis of purified 32P-labeled Gadd45 fusion protein. This protein was used to detect filter bound PCNA protein, while filter bound Gadd45 protein could also be detected by free PCNA molecules. Using recombinant proteins in conjunction with immunoprecipitation and immunoblotting, we show that Gadd45 competes with p21 for binding to PCNA and conversely, p21 blocks the ability of Gadd45 to bind PCNA. In addition, p21 appears to disrupt PCNA trimers whereas Gadd45 has a lesser effect. PCNA trimer disruption was also observed in UV-irradiated cells but not in repair-defective xeroderma pigmentosum group A (XP A) cells. PMID- 7478511 TI - Induction of Fas-mediated apoptosis in p53-transfected human colon carcinoma cells. AB - To investigate the biological function of p53 in colon carcinoma cells, a wild type p53 expression plasmid under the control of the human cytomegalovirus promoter was stably transfected into the human colon adenocarcinoma cell line WiDr, which carries a mutation of the p53 gene at codon 273. Exogenous wild-type p53 transcripts were detected at various expression levels in 8 of 117 G418 resistant clones. The growth rates of the wild-type p53+ clones in culture did not change significantly. The efficiency of colony formation in soft agar, however, was completely suppressed in two wild-type p53+ clones. This is the first to demonstrate the feasibility of stable transfection of the wild-type p53 gene under the control of non-inducible promoter in human colon cancer cells. The major alteration found was that wild-type p53+ cells which were incubated with anti-Fas IgM showed marked cytolysis with preferential over-expression of wild type p53 accompanied by overexpression of a cyclin-dependent kinase inhibitor, WAF1, whereas the endogenous mutant p53 retained its expression level. The findings suggest that a Fas-initiated pathway is incidentally linked to a p53 dependent apoptotic pathway through the reconstituted wild-type p53 gene in WiDr cells. This model should help elucidating the additional role of the p53 tumor suppressor gene and the mechanism of apoptosis in colon carcinoma cells. PMID- 7478512 TI - Differential effects of UV-B and UV-C components of solar radiation on MAP kinase signal transduction pathways in epidermal keratinocytes. AB - Exposure to solar ultraviolet (UV) light is a major cause of skin cancer, the most common human neoplasm. The earth's upper atmosphere absorbs the high energy UV-C wavelengths (100-280 nm), while allowing transmission of UV-B (280-320 nm) and UV-A (320-400 nm). It is therefore UV-B and to some extent UV-A, that contributes to most human skin malignancies. We report that the exposure of cultured keratinocytes or skin to UV-C radiation causes activation of MAP kinases (ERK and JNK). In contrast, the solar radiation associated with skin cancer (UV B) was an ineffective activator of the ERK and JNK signal transduction pathways. Therefore, while exposure of epidermal cells to UV-C radiation under laboratory conditions causes marked activation of MAP kinase signal transduction pathways, only a low level of MAP kinase signaling is involved in the response of skin to biologically relevant solar radiation. PMID- 7478513 TI - Regulation of the Src tyrosine kinase and Syp tyrosine phosphatase by their cellular association. AB - The specific activity of the Src tyrosine kinase is elevated in human colon carcinoma cells. To identify Src-binding proteins that might upregulate Src activity in these cells, a human colon carcinoma lambda gt11 expression library was screened with purified, 32P-labeled Src. The SH-PTP2 (Syp) tyrosine phosphatase was isolated and shown to associate with Src. In vitro studies demonstrated that: (1) transforming F527 Src phosphorylates Syp, and (2) Syp dephosphorylates Src at Tyr 527. Both events are known to upregulate enzyme activity. Others have shown that overexpression of the receptor tyrosine phosphatase alpha in rat embryo fibroblasts results in Src activation by dephosphorylation of Tyr 527, cell transformation and tumorigenesis. Thus, transmembrane tyrosine phosphatases may be involved in cell transformation exerting at least some of their effects through activation of Src. To the best of our knowledge, this is the first identification of an intracellular tyrosine, phosphatase which may activate Src by a similar mechanism. PMID- 7478514 TI - Mouse and rat B-myc share amino acid sequence homology with the c-myc transcriptional activator domain and contain a B-myc specific carboxy terminal region. AB - B-myc is a member of the myc gene family. Previous studies indicate that the rat B-myc gene contains a single exon which shows 77% nucleotide homology to the second exon of the rat c-myc gene. Its open reading frame (ORF) encodes a polypeptide with a predicted molecular weight of 20 kD. We have isolated a new, larger rat B-myc genomic clone. Sequence analysis of this clone confirmed the presence of one single coding exon. Furthermore, a genomic mouse B-myc clone was identified and compared to the rat homolog. Nucleotide analysis of B-myc coding and non-coding sequences suggests that it may be a functional gene evolved by selective duplication of part of the second c-myc exon. Analysis of the rodent B myc open reading frames revealed two in-frame amino acid duplications in mouse B myc and a 96% conservation at the amino acid level. Both rat and mouse B-myc proteins contain an identical and unique stretch of 14 carboxy terminal amino acid residues not found in other myc proteins. In vitro translation of rat and mouse B-myc ORF's yielded proteins that migrated as 26 kD bands in SDS-PAGE and could be immunoprecipitated by a polyclonal panmyc antiserum. Immunostaining of human lymphoma cells transiently transfected with a B-myc expression vector showed that the protein was mainly localized to the nucleus. PMID- 7478515 TI - Mapping of the interaction sites of the growth suppressor protein p53 with the regulatory beta-subunit of protein kinase CK2. AB - p53 plays an essential role in cellular growth control. Some of its distinct biological functions are regulated by interaction with cellular proteins. We have previously (Wagner et al., 1994) shown that p53 binds to the regulatory subunit of protein kinase CK2. Using C-terminal protein fragments of p53 we now demonstrate that the region between amino acids 287 and 340 on the polypeptide chain of p53 is critical for binding of p53 to the beta-subunit of CK2. Neither phosphorylation at the p34cdc2 site (aa315) nor at the CK2 site (aa392) is necessary for binding of p53 to the beta-subunit of CK2. Using deletion mutants of the beta-subunit of CK2 we also show that an internal region between amino acids 72 and 149 of the beta-subunit of CK2 is necessary for binding to p53. Thus, this study defines new functional regions on the polypeptide chains of p53 and of protein kinase CK2. PMID- 7478516 TI - Expression of constitutively activated hepatocyte growth factor/scatter factor receptor (c-met) in B16 melanoma cells selected for enhanced liver colonization. AB - The murine melanoma B16-LS9 cell line was obtained after repeated passages in vivo through the liver of syngeneic mice, and shows an enhanced ability to colonize the liver after intravenous inoculation when compared to its parental, unselected counterpart B16-F1. We have previously shown that paracrine growth effects mainly account for better growth of B16-LS9 in the liver than at other sites, and more recently we reported hepatic transferrin as a factor contributing to efficient growth in the liver. Here we show that increased expression of constitutively activated c-met (the receptor for Hepatocyte Growth Factor/Scatter Factor) consistently occurs during selection of B16 cells through the liver. Constitutive activation of c-met seems to follow its own overexpression and not to depend on an autocrine mechanism. As a consequence, liver-selected B16 melanoma cells have higher tyrosine-kinase activity and higher amounts of tyrosine-phosphorylated proteins than parental B16-F1 or lung-specific B16-F10 cells. Overexpression of constitutively activated c-met enhances motility and invasiveness of B16-LS9 cells, presumably favoring their colonization efficiency in vivo. However, whether levels of c-met expression also determine the organ specificity of B16 melanoma cells needs further clarification. PMID- 7478518 TI - Suppression of tumorigenicity in three different cell lines of human oral squamous cell carcinoma by introduction of chromosome 3p via microcell-mediated chromosome transfer. AB - It has been suggested that loss of the short arm of chromosome 3 is one of the most frequent abnormalities in human head and neck cancers including oral squamous cell carcinomas (SCC) and that one or more putative tumor suppressor gene(s) which may contribute to the initiation and/or progression of these tumors might be located on chromosome 3p. In this study, we examined the effects of introducing human chromosome 3 or 7 by microcell hybridization on the tumor associated phenotypes of three different human oral SCC cell lines, HSC-2, HSC-3 and HSC-4. Transfer of a single chromosome 3p completely suppressed the tumorigenicity of all three parental cell lines, which showed a significant decrease in growth rate in vitro and morphological changes. In contrast, transfer of chromosome 7 had no effect on HSC-2 and HSC-4 cells, although it suppressed the tumorigenicity of HSC-3 cells without modifying their in vitro growth properties. Our findings provide additional confirmatory evidence that loss or inactivation of putative tumor suppressor gene(s) present on chromosome 3p might be primarily involved in the development of human oral SCC. The possibility that chromosome 7 may carry another tumor suppressor gene(s) is also discussed. PMID- 7478517 TI - Mesodermal defect in late phase of gastrulation by a targeted mutation of focal adhesion kinase, FAK. AB - FAK is a unique non-receptor protein tyrosine kinase that was found in cellular focal adhesions. An increasing number of in vitro observations has suggested that FAK mediates signaling through integrins brought about by interactions with extracellular matrix (ECM). It is highly tyrosine-phosphorylated in v-src transformed cells and during embryogenesis. To clarify the function of FAK in cell-ECM interactions, embryonic phenotype of its mutant was analysed. FAK deficient embryos could implant and initiate gastrulation normally, but showed abnormalities in subsequent development. The abnormalities were characterized as a general deficiency in mesoderm, and the phenotype was quite similar to that caused by fibronectin-deficiency. The results suggest that FAK mediates fibronectin-integrin interactions uniquely at this stage of development, thereby playing an essential role in development of mesodermal cell lineages. PMID- 7478519 TI - The Wnt-1 proto-oncogene regulates MAP kinase activation by multiple growth factors in PC12 cells. AB - PC12/Wnt-1 cells display morphological changes in response to stimulation by select growth factors but do not respond to NGF. Furthermore, stimulation by EGF can induce neuronal differentiation in these cells but not in wild type cells. We have found that in these cells, compared to wild type PC12 cells, FGF and EGF stimulation of MAP kinase activity is enhanced, while NGF stimulation of MAP kinase in diminished. Finally, in cells expressing Wnt-1, the effect of cyclic adenosine monophosphate (cAMP) on MAP kinase activation is reversed; cAMP stimulates MAP kinase in wild type PC12 cells but inhibits MAP kinase in PC12/Wnt 1 cells. These data suggest that Wnt-1 expression alters the specificity of growth factor signaling in neuronal cells. PMID- 7478520 TI - Biochemical and mutagenic analysis of the melanoma tumor suppressor gene product/p16. AB - P16 was originally discovered by its ability to interact with CDK4 and to specifically inhibit the catalytic activity of the CDK4/D1 kinase. Increased attention has focused on the p16 gene because of its location on chromosome 9p21, a region involved in chromosomal rearrangements in a large number of tumor types. The p16 gene is also mutated in a large number of tumor cell lines and primary tumor cells. Furthermore, linkage analysis studies suggest that the p16 gene is involved in familial melanoma susceptibility. Due to the oncogenic potential of mutations in this tumor suppressor, it is important to identify and characterize those mutations which alter p16 activity. We have performed a systematic analysis of melanoma associated p16 mutants and of mutants generated in charge to Ala mutagenesis. Using microtiter plate assays to measure both p16-cdk4 binding and cdk4/D1 kinase activity, we show here that the melanoma associated mutants are defective, as are some of the Ala mutants. These results support the idea that p16 mutation, via its deregulation of the cdk4/D1 pathway, is of biological significance in the development of melanoma. Furthermore, we have defined a region within the p16 molecule in which changes are likely to result in a defective protein. PMID- 7478521 TI - Increased levels of p21WAF1/Cip1 in human brain tumors. AB - The cdk inhibitor p21WAF1/Cip1 (p21), which can be transcriptionally activated by p53, functions to block cell cycle progression. In this study, we analysed the expression of p21 in normal and reactive brain and in gliomas of various malignancy grades. Southern blotting showed no p21 gene deletion. Western blotting and immunohistochemical assay showed that the levels of p21 protein in normal and reactive brain tissue were very low; however, p21 was elevated in a majority of gliomas tested, regardless of their malignancy grades. In glioblastoma multiforme, marked elevation of p21 was observed in samples harboring either wild-type or mutant p53. But, in anaplastic astrocytomas, the level of p21 was not elevated in samples harboring mutant-type p53. Immunohistochemical staining of paraffin-embedded astrocytomas and glioblastomas showed that tumor cells and not contaminating normal cells were positive for p21. Therefore, overexpression of p21 appears to be an early event in the development of glial neoplasms and p53-dependent p21 expression appears to be tumor grade specific. PMID- 7478522 TI - A mutated p53 gene alters thyroid cell differentiation. AB - p53 is the gene most frequently found mutated in human neoplasias. In the majority of tumors, p53 mutations contribute to the progression towards stages of increasing malignancy with the appearance of an undifferentiated phenotype. Also in thyroid cancerogenesis, p53 mutations correlate with the loss of the differentiated phenotype. The results presented here, suggest a direct involvement of p53 in the molecular mechanisms regulating cellular differentiation in thyroid since a mutated p53 gene markedly affects the growth potential and differentiated functions of the rat thyroid cell line PC Cl 3. Blockage in the expression of the PAX-8 transcription factor seems to be a key event in the loss of thyroid differentiated functions induced by the mutated p53 gene. Thyroid cells carrying a mutated p53 gene did not form colonies in soft agar or tumors in athymic mice, suggesting that a mutation of the p53 gene is not sufficient for the induction of the malignant phenotype and probably a cooperation with other oncogenes is necessary to accomplish full malignancy. No effect on either growth or differentiation of thyroid cells was exerted either by overexpression of the wild-type p53 gene, or by the vector alone. PMID- 7478523 TI - Characterization of RET proto-oncogene 3' splicing variants and polyadenylation sites: a novel C-terminus for RET. AB - The RET proto-oncogene, which encodes a receptor tyrosine kinase, displays multiple alternative splicing variants. Splicing of sequences 3' of exon 19 to generate several coding and untranslated region (UTR) sequences has been previously reported. We have sequenced the full length RET coding region and characterized the transcripts and 3' UTRs generated by alternative splicing of the RET 3' terminus. These analyses were performed using both RET cDNA cloned from a pheochromocytoma library and reverse transcriptase PCR products generated using RNA from a neuroblastoma cell line (LA-N-2). Three different carboxyl termini were identified. In addition to the nine and 51 terminal amino acid forms already known, we identified a third with 43 terminal amino acids predicted to encode a novel RET protein isoform. A total of 3621 base pairs of DNA 3' of exon 19, which spans the alternatively spliced exons and RET UTRs, was sequenced. Four polyadenylation sites were identified. The observed combinations of polyadenylation sites and 3' coding sequence suggest that RET transcripts with up to 10 different 3' sequences and up to 40 different full length RET transcripts may exist. PMID- 7478524 TI - Phorbol ester-resistant monoblastoid leukemia cells with a functional mitogen activated protein kinase cascade but without responsive protein tyrosine phosphatases. AB - Human monoblastoid leukemia U937 cells differentiate to monocyte/macrophage upon treatment with phorbol ester, 12-o-tetradecanoylphorbol-13-acetate (TPA). Previous studies, including our own, have demonstrated that drug-induced differentiation of leukemia cells is associated with genetic and enzymatic activations of protein tyrosine phosphatases (PTPases). In this study, to further investigate a relationship between PTPase activation and leukemic differentiation, we established TPA-resistant U937 variant UT16 cells. Unlike known TPA-resistant cells whose resistance is mainly due to lack or down modulation of protein kinase C (PKC), UT16 cells showed TPA-induced activation of PKC, Raf-1, and ERK/MAP kinases similar to the parental U937 cells. Interestingly, however, UT16 cells exhibited altered binding activity of AP-1 complexes, decreased ability to induce c-jun and c-fos gene expressions, and failure to differentiate to a monocytic lineage. Based on these observations, UT16 cells could be considered a novel type of TPA-resistant cell. Among UT16 cells, most of TPA-inducible PTPase genes, PTP-1C, PTP-MEG2, P19-PTP, HPTP epsilon, and PTP-U1, did not respond to TPA. Consistently, TPA increased PTPase enzymatic activity in U937 but not in UT16 cells. Taken together, activation of PTPases is well correlated with TPA-induced differentiation of U937 cells. These findings indicate that gene expression and enzymatic activity of some PTPase isozymes described here are regulated by a TPA-mediated signaling event and are likely to be used as biomarkers for the monocytic differentiation of myeloid leukemia cells. PMID- 7478525 TI - Characterization of essential domains for the functionality of the MHBst transcriptional activator and identification of a minimal MHBst activator. AB - Integrated hepatitis B virus DNA derived from hepatocellular carcinomas can express, in one third of the cases investigated so far, a transcriptional activator encoded from 3' terminal truncated surface (preS/S) genes resulting in a C-terminally truncated middle surface protein (MHBst). Since MHBst, in contrast to the secreted MHBs, is retained in the secretory pathway at the ER, the question as to whether the retention generates the transcriptional activator function was investigated. Through fusion of MHBs to the ER-retention signal KDEL, it was shown that the intracellular retention does not generate the transcriptional activator function. Tryptic digestions of microsomal vesicles revealed that the amino terminal domain of MHBst directs into the cytoplasmic compartment, whereas in MHBs this domain directs into the lumen of the ER. This structural difference appears to be why transcriptional activator function arises. Through deletion analysis it was shown that non-membrane-associated MHBst proteins are also functional activators. Nonmembrane associated MHBst proteins represent a second class of MHBst proteins. These MHBst-proteins are homogenously distributed all over the cell and show no difference in functionality as compared to the membrane-associated MHBst proteins. MHBst53 (truncated at aa53) was shown to be a minimal activator of this class. Both classes of MHBst proteins were found to form dimers; an which is involved in mediating the dimerization. The integrity of this domain was also revealed to be a prerequisite for the functionality of the activator, suggesting a linkage between dimerization and functionality. PMID- 7478526 TI - Expression of Mts1, a metastasis-associated gene, increases motility but not invasion of a nonmetastatic mouse mammary adenocarcinoma cell line. AB - The mts1 gene codes for a 101 amino acid protein belonging to the S100 subfamily of Ca(2+)-binding proteins. Mts1 is overexpressed in metastatic cancers as compared to their nonmetastatic counterparts, and although mts1 is known to be involved in the metastatic phenotype (Davies et al., 1993; Grigorian et al., 1993), the role mts1 plays in this process is not clearly understood. In order to determine what role mts1 plays in the process of metastasis, we have performed transfection studies on nonmetastatic and metastatic mouse mammary adenocarcinoma cell lines, CSML0 and CSML100, respectively (Senin et al., 1983, 1984). The metastatic variant, CSML100, expresses high levels of mts1, whereas the nonmetastatic variant, CSML0, expresses almost no mts1. CSML0 cells transfected with mts1 were assessed in in vitro motility and invasion assays, as well as in vivo metastasis assays to determine the role of mts1 in these processes. Cell lines expressing mts1 display an altered morphology as well as increased motility in modified Boyden chemotaxis chambers. However, no significant increase in in vitro invasion or in in vivo metastasis was observed. Therefore, the presence of mts1 may be important for metastasis by increasing motility, but may not be sufficient for invasion in vitro or metastasis in vivo. Very low levels of type IV collagenase activities were observed in CSML0 cells and the transfectants, as opposed to the highly metastatic CSML100 cells, where high levels of type IV collagenase activities were observed. It is possible that the presence of these proteases in addition to mts1 may be responsible for the high metastatic potential of the CSML100 in vivo. PMID- 7478527 TI - A novel cytoplasmic substrate for cdk4 and cdk6 in normal and malignant epithelial derived cells. AB - Cyclins and cyclin-dependent kinases (cdk) have been identified as important regulators of cell replication. Molecular alteration in the cdk pathways appear to be important in cancer with some cyclins (eg cyclin D and E) proposed to be oncogenes and some inhibitors of cdk (eg p16) proposed to be tumor suppressor genes. In human breast carcinoma cell line MDA361 both cyclin D and E are overexpressed and cdk 4 and 6 are the predominate kinases which phosphorylate retinoblastoma protein and to a greater extent a novel 88 kDa protein. This 88 kDa protein was detected as a significant substrate in five of seven breast carcinoma cell lines, three lung carcinoma cell lines as well as in primary breast and lung epithelium. Normal human and murine T lymphocytes and established lymphoid cell lines are devoid of this protein and minimal amounts were detected in normal human fibroblast. In contrast to retinoblastoma protein, the 88 kDa protein appears to be more prevalent in the cytosolic than the nuclear subfraction. The phosphorylation of this 88 kDa protein by the G1 associated cdks suggest that this protein may represent another targeted substrate regulating the G1 phase of the cell cycle. PMID- 7478528 TI - Cloning, characterization, and differential expression of MDK2 and MDK5, two novel receptor tyrosine kinases of the eck/eph family. AB - Using a polymerase chain reaction-based strategy for the cloning of developmentally regulated receptor tyrosine kinases, we identified two novel members of the eck/eph-related subfamily which, in analogy with the recently identified mouse developmental kinase 1 (MDK1), were designated MDK2 and MDK5. MDK2 is highly homologous to the mouse kinase Myk-1 and the human kinase Htk, whereas MDK5 represents the mouse homologue of human Hek2. Northern blot analyses of adult mouse tissues revealed a 4.7 kb transcript of MDK2 and a 4.8 kb transcript of MDK5 in various organ systems, including lung, liver, kidney, intestine, muscle, heart, and, in the case of MDK5, also the brain. In addition to the full-length transcripts, smaller fragments were identified that probably represent truncated receptors. Northern blot analysis and in situ hybridization of mouse embryos indicated abundant expression during embryonic development, with preferential involvement of tissues of epithelial and endothelial origin for both kinases and of the spinal cord gray matter for MDK5. Unlike most other members of the eck/eph-related subfamily, the expression of MDK2 and MDK5 is not primarily restricted to neuronal structures, and their abundant presence in various organ systems during embryonic development suggests an important role in gestational growth and differentiation. PMID- 7478529 TI - GRB2 and SH-PTP2: potentially important endothelial signaling molecules downstream of the TEK/TIE2 receptor tyrosine kinase. AB - TEK is a newly cloned receptor tyrosine kinase that is expressed predominantly in the endothelium of actively growing blood vessels. Disruption of TEK function in transgenic mice results in a profound defect in vascular development leading to embryonic lethality. These studies show that TEK signaling is indispensable for the development of the embryonic vasculature and suggest that TEK signaling may also be required for the development of the tumor vasculature. Because the ligand for TEK has not been identified, it has been difficult to study signal transduction by this important endothelial receptor. To circumvent this problem, a soluble TEK kinase domain (GTEKH) was developed which could be easily purified, autophosphorylated, and radiolabeled. Using the autophosphorylated, radiolabeled GTEKH to probe a mouse embryo expression library only two candidate signaling molecules were isolated, SH-PTP2 and GRB2. Autophosphorylated GTEKH associated with GRB2 and SH-PTP2 from endothelial lysates and not with PI3 kinase or PLC gamma. The association of GRB2 and SH-PTP2 with TEK was highly dependent on specific tyrosine residues in the TEK c-tail. These studies identify GRB2 and SH PTP2 as potentially important mediators of TEK signaling that may trigger crucial endothelial responses during embryonic vascular development and during pathologic vascular growth. PMID- 7478530 TI - Raf-1 kinase and ERK2 uncoupled from mitogenic signals in rat fibroblasts. AB - The MAP kinase pathway impinging on ERK2 has been shown to be integrally associated with mitogenic signalling in many cell types. Previously, we and others have demonstrated that oncogenic forms of Raf-1 kinase, when expressed in fibroblasts, lead to the constitutive activation of ERK2, the de-regulation of c fos expression and increased cell proliferation. Here we describe an exception to this scenario. In Rat6 cells, although both ERK1 and ERK2 are activated in response to mitogens that induce c-fos expression, such as Epidermal Growth Factor (EGF), lysophosphatidic acid (LPA) or serum, expression of v-Raf fails to induce c-fos expression and increase proliferation. However, ERK2 is activated by v-Raf expression. The co-transfection of an interfering mutant of ERK2 has no effect on the level of c-fos reporter expression in Rat6 cells whereas the analogous ERK1 mutant reduces its expression. Furthermore, the spontaneous focus formation observed in Rat6 cells is susceptible to the interfering mutant of ERK1 but resistant to that of ERK2. Thus, not only do mitogenic signals appear to by pass both Raf-1 kinase and ERK2, the Raf-1-ERK2 pathway seems to be functionally compromised in Rat6 cells as its activation leads neither to c-fos expression nor to increased proliferation. PMID- 7478531 TI - Regulation of c-Myb through protein phosphorylation and leucine zipper interactions. AB - c-Myb function is modulated in part by a negative regulation domain which encompasses a leucine zipper (LZ). When E. coli-expressed c-Myb with wild type or mutated LZ proteins are assessed for DNA binding activity, the mutant form is substantially better at DNA binding than the wild type (WT) form. In contrast, the DNA binding activity of the WT protein is increased to an equivalent level of activity of the LZ-mutant when both are expressed in rabbit reticulocyte lysates (RRL) or insect cells. The possibility that phosphorylation overcomes the negative influence of the LZ was investigated. E. coli-expressed mutant, but not wild type c-Myb proteins, were shown to be substrates for Casein Kinase II (CKII) and cAMP-dependent Protein Kinase (PKA). The phosphorylation sites for CKII and PKA were serines 11 and 12, and 8 and 116, respectively. Serines 11 and 12 were found to be phosphorylated in recombinant wild type and mutant c-Myb expressed in insect cells and DNA binding was markedly reduced following phosphatase treatment. Substitution of serines 11 and 12 with glutamic acid and alanine in E. coli-expressed Myb demonstrated that these amino terminal residues influence the negative effect on DNA binding exerted by the LZ. Collectively, these observations support the notion that phosphorylation of serines 11 and 12 positively modulate DNA binding. PMID- 7478532 TI - Homozygous deletions at 8p22 and 8p21 in prostate cancer implicate these regions as the sites for candidate tumor suppressor genes. AB - Frequent loss of an allele at specific chromosomal regions implicates these regions as sites of tumor suppressor genes (TSG) that become inactivated during tumor progression. We have studied chromosome 8p allele losses in 32 primary human prostate carcinomas with 16 polymorphic microsatellite sequences. Overall, 22 of 32 (69%) informative specimens showed loss of allele in at least one locus. The most frequent losses of heterozygosity (LOH) occurred at the LPL locus (46%) on chromosome 8p22 and at the D8S360 (45%) and NEFL (43%) loci on chromosome 8p21. Homozygous deletions were detected at the LPL and NEFL loci at 8p22 and 8p21, respectively. The minimal region with frequent LOH and homozygous deletion, around the LPL locus, was restricted between the MSR locus and the D8S258 marker, separated by less than 9 cM. The second region was restricted between markers D8S1128 and D8S131 separated by 12 cM. The results suggest the existence of two chromosome 8p sites for candidate TSGs in prostate cancer. PMID- 7478533 TI - c-fos mRNA instability determinants present within both the coding and the 3' non coding region link the degradation of this mRNA to its translation. AB - The instability of oncogenic mRNA such as c-fos mRNA is controlled in cis by sequences present in both the coding and the 3' untranslated regions (3'UTR). The latter contains AU-rich elements (ARE) which, depending on the cellular context, mediate either their rapid degradation or inhibit their translation. These observations, along with the known increase of the life spans of many unstable mRNA promoted by inhibitors of protein synthesis, raise the possibility that both processes are linked. To investigate further the putative involvement of translation in both coding region and ARE-mediated rapid decay of c-fos mRNA, we designed an expression vector based on the use of the ferritin mRNA iron regulatory element (IRE). The latter structure links translation to intracellular iron concentration when inserted at the proper location within the 5'UTR. Rapid degradation of a beta-globin/c-fos 3'UTR construct was prevented by Desferrioxamine, an iron chelator, and facilitated by ferric ammonium citrate or hemin, while stability of other mRNAs not containing the IRE or the ARE were unchanged. The same conclusion was reached when the stability of a c-fos mRNA devoid of ARE was assessed in function of iron availability. PMID- 7478534 TI - ETS1 transactivates the human GM-CSF promoter in Jurkat T cells stimulated with PMA and ionomycin. AB - Activation of T helper cells results in coordinate expression of a number of cytokines involved in differentiation, proliferation and activation of the haematopoietic system. Granulocyte-macrophage colony-stimulating factor (GM-CSF) is one such cytokine whose increased expression results partly from increases in transcription. Cis-acting elements with NF kappa B, AP-1 and ETS-like motifs have been identified in the promoter region of the GM-CSF gene, which are important for transcriptional activity following PMA and ionomycin stimulation. A number of the ETS family of transcription factors are expressed in T cells, including ETS1 and ELF1. Here we describe the ability of these factors to interact with a site (GM5), located within the CLE0 element, -47 to -40 upstream of the GM-CSF transcription initiation site. Exogenous ETS1, but not ELF1, can transactivate GM CSF, through the GM5 site, in a PMA/ionomycin dependent manner. Other unidentified ETS-like factors present in Jurkat cells are also capable of binding GM5. Mutation of the core ETS binding site from -GGAA- to -GGAT- prevents the binding of ETS-like factors with the exception of ETS1. The GM-CSF promoter, modified in this way to be ETS1 specific, is fully responsive to PMA/ionomycin induction, in addition to ETS1 transactivation in the presence of PMA and ionomycin. Together these data suggest that ETS1 may be involved in mediating the increased GM-CSF production associated with T cell activation. PMID- 7478536 TI - Ultraviolet B irradiation-induced G2 cell cycle arrest in human keratinocytes by inhibitory phosphorylation of the cdc2 cell cycle kinase. AB - In response to genotoxic stress, cell cycle progression can be arrested at certain checkpoints which serve to maintain genomic integrity. We have investigated the mechanism of ultraviolet B (UVB) irradiation-induced cell cycle arrest in normal human keratinocytes and in the HaCaT keratinocyte cell line which carries mutant p53 tumour suppressor protein. While only normal keratinocytes showed a delay in G1 following sublethal UVB irradiation both cell types exhibited prolonged G2 arrest attributable to rapid inhibition of cyclin B associated cdc2 kinase activity. This inhibition coincided with increased tyrosine phosphorylation of cdc2 and was reversed by the cdc25C phosphatase in vitro. The data indicate that UVB-induced G2 arrest in mammalian cells is mediated by inhibitory tyrosine phosphorylation of cdc2 and acts as a defense mechanism against DNA damage irrespective of the cells' p53 status. PMID- 7478535 TI - Homozygous deletion of the MTS1/p16 and MTS2/p15 genes and amplification of the CDK4 gene in glioma. AB - Loci on chromosome 9p are frequently deleted in several malignant tumors, suggesting the presence of putative tumor suppressor genes. The MTS1/p16 and MTS2/p15 genes on 9p are considered to be candidates. Binding of p15 and p16 cell cycle-regulatory proteins to the cyclin dependent protein kinase CDK4 inhibits CDK4/cyclin D dependent phosphorylation of retinoblastoma protein. We analysed the DNAs from 37 gliomas of several grades of malignancy for allelic loss of chromosome 9p and aberrations of the MTS1/p16 and MTS2/p15 genes. We detected losses of one allele and homozygous deletions at loci, including those of the MTS1/p16 and MTS2/p15 genes, in 10 and 3 tumors, respectively. However, we did not detect any tumor-specific mutation in the two genes. The CDK4 gene was amplified in two malignant gliomas without homozygous deletion of the MTS1/p16 and MTS2/p15 genes and one malignant glioma with an allelic loss of the genes. These data suggest that aberrations of the genes coding for components of the cell cycle-regulatory system occurred in at least 15 of 37 gliomas. PMID- 7478537 TI - The POU domain transcription factor Brn-2: elevated expression in malignant melanoma and regulation of melanocyte-specific gene expression. AB - Previous work has shown that melanoma cell lines express a distinct octamer binding protein. Given the role of octamer-binding proteins in cell differentiation and development, the role this factor is a key issue in understanding melanocyte differentiation and transformation. Using a proteolytic clipping assay, we show that the melanoma-specific octamer factor is Brn-2/N Oct3, a POU domain protein previously known to be expressed in adult brain and in the developing nervous system. N-Oct3 mRNA was detected in a range of human melanoma cell lines and was around 10-fold elevated compared to normal human melanocytes while mRNA for Brn-2 was also detected in a mouse melanoblast cell line. Expression of Brn-2/N-Oct3, in melanoma cells in cotransfection assays activated the expression of the MHC class II DR alpha promoter but repressed the activity of the melanocyte-specific tyrosinase promoter. Repression correlated with Brn-2/N-Oct3 binding in a mutually exclusive fashion with basic-helix-loop helix-leucine-zipper (bHLH-LZ) transcription factor USF in vitro and with Brn-2 expression preventing activation of the tyrosinase promoter by the bHLH-LZ factor Microphthalmia in vivo. The potential role of Brn-2/N-Oct3 in melanocyte differentiation and gene expression is discussed. PMID- 7478538 TI - Survival and death of prelymphomatous B-cells from N-myc/bcl-2 double transgenic mice correlates with the regulation of intracellular Ca2+ fluxes. AB - Coexpression of the proto-oncogenes c-myc and bcl-2 under the control of the immunoglobulin enhancer E mu provokes the rapid development of primitive lymphoid tumors in transgenic mice. In the present study we show that the myc family members N-myc and L-myc also cooperate with bcl-2 in oncogenesis and can provoke the development of more mature pre-B, B and T cell type lymphomas. The analysis of prelymphomatous B-cells from single E mu N-myc and bcl-2-Ig transgenic animals and from young, tumor free, double transgenic E mu N-myc/bcl-2-Ig mice revealed that E mu directed expression of N-myc leads to very rapid apoptosis after explantation and culturing compared to B-cells from normal mice. As expected, B cells from bcl-2-Ig transgenics were protected to a certain degree from apoptosis. Strikingly however, B-cells from E mu N-myc/bcl-2-Ig double transgenic animals were found to be almost completely resistant towards a number of different apoptotic stimuli. Furthermore, after treatment with H2O2, which can trigger apoptosis, B-cells from E mu N-myc animals reach levels of intracellular free Ca2+ concentrations that are comparable to B-cells from normal mice, whereas B-cells from bcl-2-Ig or E mu N-myc/bcl-2-Ig double transgenic mice show no increase in intracellular Ca2+ concentrations after stimulation with H2O2. These findings suggest that the prevention of apoptosis conferred by bcl-2 correlates with the inhibition of intracellular Ca2+ fluxes whereas induction of apoptosis mediated by N-myc requires normal Ca2+ levels. We hypothesize therefore that the regulation of intracellular Ca2+ concentrations represent one important parameter in the oncogenic cooperation between bcl-2 and N-myc. PMID- 7478539 TI - UREB1, a tyrosine phosphorylated nuclear protein, inhibits p53 transactivation. AB - Tumor suppressor p53 is a transcription activator that upregulates target genes containing the p53 binding site. UREB1, a DNA binding protein that is tyrosine phosphorylated in vivo, shares a significant homology with the human papilloma virus E6 associated protein (E6-AP). E6-AP forms a ternary complex with E6 and p53 and participates in the ubiquitination of p53. Based on the homology with E6 AP, but taking into account the nuclear localization of UREB1 and its smaller size, the present study used a transient transfection system to examine whether UREB1 influenced p53-stimulated transcription. Co-transfection of a vector expressing wildtype UREB1 with one expressing p53 into H1299, a p53 negative cell line, resulted in a pronounced suppression of p53 transactivation. The inhibitory effect was significantly attenuated by mutation of a tyrosine residue in the consensus tyrosine phosphorylation sequence of UREB1. These data suggest that optimal suppression of p53 transactivation requires tyrosine phosphorylated UREB1 and that tyrosine phosphorylation and dephosphorylation processes may be involved in the regulation of p53 transactivation. PMID- 7478540 TI - Colon carcinoma kinase-4 defines a new subclass of the receptor tyrosine kinase family. AB - Complementary DNA (cDNA) encoding a novel member of the receptor tyrosine kinase (RTK) family has been isolated from colon carcinoma tissue. Colon carcinoma kinase 4 (CCK-4) mRNA is highly expressed in human lung tissue and at lower levels in the thyroid gland and ovary. While no mRNA was found in human adult colon tissues, expression varied remarkably in colon carcinoma-derived cell lines. CCK-4 cDNA encodes a chicken KLG-related, 1071 amino acid-long transmembrane glycoprotein containing several genetic alterations within the RTK consensus sequences. These define CCK-4 as a catalytically inactive member of the RTK family of proteins and, in analogy to HER3, suggest a potentially tumor characteristic role as a signal amplifier or modulator for an as yet unidentified kinase-competent partner. PMID- 7478541 TI - The control of apoptosis and drug resistance in ovarian cancer: influence of p53 and Bcl-2. AB - Modulation of apoptosis may influence resistance to chemotherapy and therefore affect the outcome of cancer treatment. Ovarian cancer, one of the most fatal malignancies in women, is often associated with drug resistance but the cellular pathways contributing to this effect remain obscure. We have found that Bcl-2 and p53, two proteins implicated in the control of apoptosis, are frequently expressed in fresh biopsies of primary ovarian carcinoma. Examination of Bcl-2 and p53 protein levels in pairs of cis-platin sensitive and resistant ovarian cell lines demonstrated that the resistant variants over-express Bcl-2 and/or p53, apparently due to progressive expansion of Bcl-2 and/or p53 positive subpopulations during the in vitro development of resistance. Exogenous expression of Bcl-2 or a temperature sensitive mutant p53 (ts p53) in the ovarian cell line A2780 resulted in protection from drug-induced apoptosis and a delay in drug-mediated S-phase arrest. Interestingly, p53 accumulation in response to DNA damage induced by different agents was significantly delayed and reduced in the Bcl-2 transfectants compared to the control A2780 line, suggesting that Bcl-2 may act upstream of the p53 pathway. Similarly, the induction of Bax mRNA and protein was also found to be delayed in the presence of Bcl-2. Overall, our data provide further evidence for cross-talk between Bcl-2, p53 and Bax and suggest that these genes are important determinants of drug-induced apoptosis thereby modulating resistance to chemotherapy. PMID- 7478543 TI - Localization of potential tumor suppressor loci to a < 2 Mb region on chromosome 17q in human prostate cancer. AB - We recently demonstrated a high frequency of loss of heterozygosity (LOH) at the D17S856 and D17S855 (within the BRCA1 gene) loci in primary prostate cancer, suggesting that the BRCA1 gene and/or other tumor suppressor gene(s) located within the interval of the D17S856 and D17S855 loci and/or within the vicinity of this interval may be important in prostate cancer (Cancer Res., 55: 1002-1005, 1995). To further define the exact boundary of the deleted region (i.e., D17S856/D17S855) and to detect other possible LOH regions on the long arm of chromosome 17, we analysed 23 matched normal and tumor DNAs with 15 polymorphic microsatellite markers spanning chromosome 17q12-21. Eleven of 22 (50%) informative tumors showed allelic deletion at one or more of the loci studied. A minimal area of LOH was identified to extend from the proximal boundary at the D17S776 locus to the distal boundary at the D17S855 locus, spanning an estimated < 2 Mb segment on chromosome 17q21. Our results suggest that a potential tumor suppressor gene(s) may reside in the < 2 Mb region centromeric (inclusive) to the BRCA1 gene and that this tumor suppressor gene(s) may be involved in the formation of prostate cancer. PMID- 7478542 TI - Spi-1 and mutant p53 regulate different aspects of the proliferation and differentiation control of primary erythroid progenitors. AB - The emergence of leukemic cells in Friend virus complex-induced erythroleukemia is associated with two recurrent genetic alterations, namely the inactivation of the p53 tumor suppressor gene and the overexpression of Spi-1, a member of the Ets family of transcriptional regulators. In order to determine the role of these genetic alterations on the proliferation and differentiation control of erythroblasts, we expressed Spi-1 and the temperature sensitive mutant p53(V135A) in avian primary erythroid progenitors. We show that enforced expression of Spi-1 in erythroblasts obtained from bone marrow cells by expression of the ts-Sea tyrosine kinase inhibits the execution of the differentiation program normally induced in these cells in response to Epo and insulin and following inactivation of ts-Sea function. In contrast, overexpression of p53(V135A) is without effect on the ability of these cells to differentiate into erythrocytes. However, expression of p53(V135A) in erythroid progenitors obtained from bone marrow cells in the presence of SCF, TGF alpha and estradiol, was found to relieve these cells from their absolute TGF alpha requirement for long term proliferation. This phenotype is dependent upon the expression of the mutant form of p53(V135A) as it is not observed at a temperature at which p53(V135A) regains wild type p53 function. Our results show that each of the genetic alterations which characterize Friend erythroleukemic cells affect in a distinct manner the proliferation and differentiation control of primary erythroid progenitors. PMID- 7478544 TI - Detailed deletion mapping of chromosome 9p and p16 gene alterations in human borderline and invasive epithelial ovarian tumors. AB - We used PCR amplification of tandem repeats to study the pattern of allelic loss in borderline and invasive ovarian epithelial tumors using 12 primer pairs to generate a detailed deletion map of chromosome 9p. In the invasive ovarian carcinomas, there were three regions displaying high frequency of loss of heterozygosity (LOH) ranging from 31-38%. In contrast, LOH was a rare event among the borderline ovarian tumors, with one region revealing a rate of 20% and the remaining regions only 0-8% LOH. Therefore, allelic loss does not seem to be important for the development of borderline ovarian tumors. We also examined p16 gene expression and mutations in ovarian cancer cell lines and invasive and borderline ovarian tumor tissues. Southern blot analysis revealed no losses of the p16 gene in either the invasive or borderline ovarian tumors. However, the ovarian carcinoma cell lines showed a 50% homozygous deletion rate. SSCP analysis detected a mobility shift in only one (borderline) tumor. Since the primary invasive ovarian tumors did not show any deletions or mutations, it appears that p16 does not play a role in the pathogenesis of these tumors. PMID- 7478545 TI - A novel insertional mutation in the TC21 gene activates its transforming activity in a human leiomyosarcoma cell line. AB - TC21 is the fourth member of the ras gene family to exhibit oncogenic activation in human tumor cells. To assess the prevalence of activated TC21 oncogenes in human tumors, we have developed sensitive single-strand conformational polymorphism (SSCP) conditions and immunological reagents for the detection of both single base alterations and/or overt overexpression in a wide spectrum of human tumor cell lines and surgical samples. In an initial examination of 33 human tumor specimens, we observed a novel nine basepair three amino acids insertion at TC21 codon 24 in one human uterine leiomyosarcoma cell line, SK-UT 1. This mutant allele when transfected into NIH3T3 cells, displayed high transforming activity comparable to that of the Leu72 oncogenic mutant identified by expression cDNA cloning from a human ovarian carcinoma cell line. Comparing the level of GTP-binding by the mutant and normal TC21 products revealed that this novel lesion increases the GTP-bound form of the TC21 molecule. These findings imply that the mechanism by which mutations activate the oncogenic properties of this ras-related molecule is analogous to that of previously known ras family members. PMID- 7478546 TI - Expression patterns of immediate early transcription factors in human non-small cell lung cancer. The Lung Cancer Study Group. AB - In 1995, there will be 172,000 new cases of lung cancer diagnosed and 153,000 deaths from this disease in the United States. While the pathogenesis of the disease process is poorly understood, a growing body of evidence suggests that abnormalities in cellular regulatory genes may play an important role in the induction, maintenance and/or progression of some tumor types. These genes include both growth promoting oncogenes as well as growth inhibitory or suppressor genes. Included among these genetic sequences are several cellular transcription factors. A group of these factors including c-jun, c-fos and EGR1 are members of a class of genes known as immediate early genes whose expression are inducible by a variety of stimuli including mitogenic and differentiation inducing growth factors, indicating a potential important role for these genes in normal growth processes. Since these genes are involved in early regulation of cellular growth properties and at least two (c-jun and c-fos) can act as oncogenes, we wished to determine whether their expression levels were altered in human non-small cell lung cancers (NSCLC) compared to normal lung tissue. To address this, Northern blot analyses were performed using c-fos, c-jun and EGR1 probes on RNA extracted from 101 NSCLC tumor specimens and adjacent uninvolved lung tissue. Analysis of this cohort revealed that 72% of the normal tissues demonstrate significantly greater expression of these transcription factors as compared to adjacent malignant tissue. Moreover, this expression pattern appeared to be coordinate for all three genes in the majority of cases. This differential expression pattern was confirmed at the protein level using an immunohistochemical approach with antibodies directed against the c-jun, c-fos and EGR1 gene products. Southern blot analyses demonstrated no gross alterations of these sequences at the DNA level, indicating that the observed differential expression pattern was not due to gross structural changes in the genes. These data suggest that down-regulation of these genes may be involved in the pathogenesis of lung cancer. PMID- 7478547 TI - Limk1 is predominantly expressed in neural tissues and phosphorylates serine, threonine and tyrosine residues in vitro. AB - We have isolated the murine Limk1 gene, which is a single copy gene located at the distal end of mouse chromosome 5. Limk1 exhibits a 95% homology to the human homologue, LIMK, which contains two LIM domains and a putative protein kinase domain. Although Limk1 and LIMK contain all motifs found in catalytic kinase domains, amino acids previously described to be diagnostic of either serine/threonine- or tyrosine-kinases are not present. It is demonstrated that GST-Limk1-fusion protein can autophosphorylate on serine, tyrosine and threonine residues in vitro and that mutation of residue D460 within the IHRDL motif abolishes kinase activity. Northern blot showed preferential expression of a 3.5 kb message in adult spinal cord and brain. In situ hybridisation confirmed high expression levels in the nervous system, particularly in the spinal cord and the cranial nerve and dorsal root ganglia. Limk1 also contains two tandem LIM domains. These zinc-finger like domains can mediate protein-protein interactions and have been described in nuclear and cytoskeletal proteins. The combination of LIM- and kinase domains may provide a novel route by which intracellular signalling can be integrated. PMID- 7478548 TI - A single ancestral gene of the human LIM domain oncogene family LMO in Drosophila: characterization of the Drosophila Dlmo gene. AB - Members of the human TTG/RBTN family, now renamed 'LMO' for LIM-only proteins, encode proteins with two tandem copies of a LIM motif. There are three members of this family, two have been isolated at the sites of chromosomal translocations in T-cell leukaemia. The function of the LIM motifs is at present unknown. We found that the LMO-2 gene is highly conserved between mammals, Drosophila and yeast. As a first step to obtain a model system for studying the function of the LIM motifs we have isolated the Drosophila homologue Dlmo. In contrast to mammals Drosophila appears to have only one lmo gene. A 2087 bp cDNA clone was isolated from a larval cDNA library, encoding a protein of 266 amino acids. A second transcript with an alternative 5' end was identified in RNA from embryos. The Drosophila lmo protein consists of two tandem copies of the conserved LIM domain characteristic of the human LMO family and an extended amino and carboxy terminus, which is not present in the human proteins. The amino acid sequence similarity with human LMO 1 and LMO-2 in LIM 1 is 79% and 69% and in LIM-2 90% and 60%, respectively. In addition a short stretch of 25 nucleotides with a homology of 83% between LMO-2 and Dlmo is found in the 3' UTR. Dlmo, like LMO-1, has an intron after the second LIM encoding region, which is not present in LMO-2. It is expressed maternally and at a high level in early embryogenesis as well as in adults. Interestingly we observed that the Dlmo protein is immunologically related to LMO-2 and can be detected by immunohistochemistry in early cellular blastoderm embryos. The gene was localised to a genetically well characterized region (17C on the X chromosome) opening the way for identification of mutations. PMID- 7478549 TI - Overexpression of retinoic acid receptor alpha suppresses myeloid cell differentiation at the promyelocyte stage. AB - Retinoic acid receptor (RAR) alpha is required to heterodimerize with retinoid X receptor (RXRs) in order to regulate myeloid differentiation. If so, it is expected that overexpression of normal RAR alpha may perturb the RAR alpha/RXR heterodimer formation and also the differentiation of myeloid cells. We have described here the morphology and the RA response of human RAR alpha cDNA transduced murine bone marrow cells using a retroviral vector. Most of RAR alpha transduced cells displayed promyelocyte like morphology and their proportion of c kit expressing population was increased remarkably compared with the control (Neor gene transduced cells). Furthermore, this morphology was observed even after these cells were brought into the semisolid culture containing IL-3 alone. Interestingly, immature RAR alpha transduced cells differentiated into mature granulocytes under the condition of the high concentration of RA(10(-6) M). We did not observe any effect of RAR alpha on monocytes. These results indicate that overexpression of normal RAR alpha is sufficient for inducing maturation arrest of myeloid cell lineage that is similar to the phenotype found in the acute promyelocytic leukemia bearing PML-RAR alpha translocation. PMID- 7478550 TI - p53 represses SV40 transcription by preventing formation of transcription complexes. AB - There is now much evidence to suggest that the p53 tumour suppressor protein functions to monitor the integrity of the genome. When DNA damage is detected, p53 suppresses cell growth to allow repair or directs the cell into apoptosis. The mechanism of action of p53 is as yet unclear but recent evidence has accumulated to suggest that p53 might act by regulating gene expression. Consistent with this model, p53 can both activate and repress a number of viral and cellular promoters. p53 has also been shown to bind to the CCAAT-binding Factor and TATA-binding protein (TBP), and there is direct evidence that p53 represses in vitro transcription by preventing TBP from binding DNA. We now provide evidence that p53 can repress transcription from the SV40 promoter by disrupting DNA/protein complexes involving transcription factor Sp1. PMID- 7478551 TI - TGF-beta 1 and cAMP attenuate cyclin A gene transcription via a cAMP responsive element through independent pathways. AB - Transforming growth factor beta (TGF-beta) is a potent inhibitor of the proliferation of many cell lines. The expression of Cyclin A is down-regulated by TGF-beta 1 in Chinese hamster lung fibroblasts and most of this effect is mediated at the transcriptional level through a cAMP-responsive element (CRE), but does not require a functional cAMP-dependent protein kinase. However, activation of the cAMP pathway in these cells gives rise to a strong inhibition of proliferation, paralleled by a down-regulation of Cyclin A promoter activity. This effect requires the integrity of the CRE, suggesting a role for CRE-binding proteins in late G1/S controls. PMID- 7478552 TI - Frequent alterations in E-cadherin and alpha- and beta-catenin expression in human breast cancer cell lines. AB - Alterations in intercellular junction and membrane cytoskeletal proteins may underlie some of the morphological, invasive and metastatic properties of cancer. E-cadherin, a transmembrane protein that functions in epithelial cell-cell interactions at adherens junctions, is linked to the membrane cytoskeletal matrix through interactions with alpha- and beta-catenin. We have carried out studies of E-cadherin and alpha- and beta-catenin in 18 breast cancer cell lines to determine the prevalence and nature of alterations in these genes in breast cancer. Ten lines failed to express E-cadherin protein at detectable levels and seven failed to produce detectable levels of E-cadherin transcripts. In a line lacking E-cadherin expression (SK-BR-3) a homozygous deletion of a large portion of the E-cadherin gene was noted. Localized sequence alterations in E-cadherin transcripts were not identified in any lines. In contrast to the results of a previous study, no relationship was identified between E-cadherin expression and HER-2/NEU expression. Two of the 18 lines had no detectable alpha-catenin protein and six others had reduced levels. The two lines lacking alpha-catenin protein had reduced or undetectable levels of alpha-catenin transcripts, while no consistent changes in alpha-catenin transcript levels were seen in the lines with reduced, but detectable, levels of alpha-catenin protein. Similarly, although two lines lacked beta-catenin protein, in most lines the levels of beta-catenin transcripts and protein were not well correlated with one another. Our findings suggest that alterations in E-cadherin and alpha- and beta-catenin expression are frequent in human breast cancer-derived cell lines, and that in some cases the decreased expression may result from mutations in the genes. Furthermore, the frequent alterations in the expression of these proteins argue that loss of function in the E-cadherin-catenin pathway may be critical in the development of many breast cancers. PMID- 7478553 TI - Downregulation of the Ras activation pathway by MAP kinase phosphorylation of Sos. AB - Formation of a complex of the nucleotide exchange factor Sos, the SH2 and SH3 containing adaptor protein Grb2/Sem-5 and tyrosine phosphorylated EGF receptor and Shc has been implicated in the activation of Ras by epidermal growth factor (EGF) in fibroblasts: related mechanisms for activation of Ras operate in other cell types. An increase in the apparent molecular weight of Sos has been reported to occur after several minutes of receptor stimulation due to phosphorylation by mitogen-activated protein (MAP) kinases. We report here that treatment of human peripheral blood T lymphoblasts with phorbol esters causes a similar shift in mobility of Sos. This modification of Sos does not alter its ability to bind Grb2, but correlates with strong inhibition of the binding of the Sos/Grb2 complex to tyrosine phosphorylated sequences, either a tyrosine phosphopeptide in cell lysates or p36 in intact cells. This effect, along with the mobility shift of Sos, can be mimicked in vitro by phosphorylation of Sos by the mitogen activated protein kinase, ERK1. A novel negative feedback mechanism therefore exists whereby activation of MAP kinases through Ras results in the uncoupling of the Sos/Grb2 complex from tyrosine kinase substrates without blocking the interaction of Sos with Grb2. PMID- 7478554 TI - HOX11 expression in pediatric acute lymphoblastic leukemia is associated with T cell phenotype. AB - Based on cytogenetic analysis, chromosomal translocations involving band 10q24 occur in 4-7% of T-cell acute lymphoblastic leukemia (T-ALL). The HOX11 gene is located in this chromosomal band and is activated by translocations t(10;14) (q24;q11) and t(7;10) (q35;q24). Ectopic expression of the HOX11 gene appears to be involved in the development of T-cell tumors. The aim of this study was to determine the frequency of HOX11 activation in pediatric ALL patients and to correlate gene expression with ALL immunophenotype. None of 53 B-lineage ALLs was positive for HOX11 expression, however, Northern blot and RT-PCR analysis revealed that four of 12 T-ALLs (33%) showed expression of the gene. In order to assess whether HOX11 expression is present in other pediatric malignancies we examined a panel of 20 tumor cell lines established from solid tumors and leukemias, but none of them showed expression of HOX11. Using our RT-PCR method we confirmed that HOX11 expression is not detectable in normal T-cells. These findings indicate that HOX11 expression in pediatric ALL is exclusive to T-ALL and does not occur in B-lineage ALL. The frequency detected by molecular techniques was significantly higher than the frequency reported in the literature based on cytogenetic analysis. These results support the notion that ectopic expression of the HOX11 homeobox gene is a crucial step in T-cell tumorigenesis. PMID- 7478555 TI - Malignant transformation of rat embryo fibroblasts by cotransfection with eleven human mutant p53 cDNAs and activated H-ras gene. AB - Eleven different missense and one nonsense mutant-type p53 cDNAs, which have been frequently detected in human colorectal carcinomas, were constructed and examined for their ability to cooperate with activated human H-ras genes, pSK2 and pHs49, in transfection of rat embryo fibroblasts (REF). Each missense mutant-type p53 cDNA with either of the two activated H-ras genes transformed REF with a different frequency of transformation depending on the different kind of mutation, whereas wild-type p53 (with ras), nonsense mutant-type p53 (with ras), as well as mutant-type p53 (alone) and ras (alone), did not transform REF. Six transformed REF cell lines were established from cotransfection with missense mutant-type p53 cDNA and ras gene; all of them exhibiting exogenous human p53 DNA, RNA, protein, and H-ras DNA and RNA. All six transformed cell lines showed both tumorigenicity and lung metastatic potential in nude mice. They also exhibited 92 kilodalton gelatinase activity, which was not detected in parental REF. These results suggest that missense mutations in p53 gene have a role in malignant transformation as well as metastatic potential. PMID- 7478557 TI - Deletion mapping defines different regions in 1p34.2-pter that may harbor genetic information related to human colorectal cancer. AB - Cytogenetic and molecular analyses of colorectal cancer cells have revealed deletions at 1p as prominent alterations, suggesting that genetic information on the short arm of chromosome 1 has a role in tumorigenesis. In this study we have used 33 microsatellite markers to fine map deletions at 1p in primary colorectal carcinomas. We found 1p-deletions in 84% of the cases (31/37). High frequencies of loss of heterozygosity (LOH), often the result of small independent interstitial deletions in the same tumor, defined three regions, that may harbor genetic information relevant for colorectal cancer: (i) region A between D1S243 and D1S468 (7cM; 1p36.3); (ii) region B between D1S436 and D1S199 (7cM; 1p35.1 36.31) and (iii) region C between D1S496 and D1S255 (1cM; 1p34.2-35). In addition we identified seven cell lines with LOH at 1p, all of which have deletions that span at least from the distal border of region A to the proximal border of region C. PMID- 7478556 TI - Selection of activating mutations of c-fms in FDC-P1 cells. AB - FDC-P1 haemopoietic cells were used to select mutations of c-fms that constitutively activate the receptor for macrophage-colony stimulating factor (M CSF or CSF-1). One mutation changed Ser 929 to Gly within a Ser/Gly rich region of the C-terminal tail and a second changed a nearby, highly conserved Leu 926 for Pro. A third mutation (D802V) changed Asp 802 to Val within the alpha L12/beta 9 region of the tyrosine kinase domain, so supporting the crystallographic evidence that this region triggers kinase activation. A c-kit mutation exactly equivalent to D802V was previously identified in a leukamic cell line and was demonstrated here to be transforming. Surprisingly, although D802V potently transformed FDC-P1 cells, it could not induce Rat-2 fibroblast foci, even in the presence of M-CSF. It is suggested that the accelerated receptor degradation induced by D802V may account for its cell specific effect. PMID- 7478558 TI - Enhanced p53 activity and accumulation in response to DNA damage upon DNA transfection. AB - In response to DNA damage the wild-type tumor suppressor protein p53 accumulates in the nucleus of rodent and primate cells. To investigate the minimal requirement for this reaction the cellular DNA was restricted by two alternative ways: (i) by calicheamicin gamma 1, an enediyne, which causes direct, sequence specific DNA damage, as shown by fluorimetric analysis of DNA unwinding and by poly(ADP-ribose) polymerase activation. The dose-dependent DNA damage correlated with the nuclear p53 accumulation. In addition, restriction was generated (ii) by the intracellular introduction of the restriction enzyme PvuII, which generates blunt-ended DNA breaks, applying a mild hypotonic shock (pellet method). Previous transfection of linear or circular, single- or ds, DNA, followed by mitomycin C treatment, lead to a dramatic increase in nuclear p53 accumulation and p53 activity according to electrophoretic mobility shift analysis. The nature of transfected DNA was irrelevant for enhanced accumulation. The data suggest, that the cellular p53 response to DNA damage is sensitized by uptake of exogenous DNA. PMID- 7478559 TI - Regulation of the c-fms promoter in murine tumour cell lines. AB - Macrophage colony-stimulating factor (CSF-1) mRNA was detected in a wide range of murine tumour cell lines. Stable transfection with a CSF-1 receptor (c-fms) expression plasmid increased the number and size of colonies formed in soft agar by tumour cell lines that were already clonogenic, but did not induce transformation in non-clonogenic lines. To identify mechanisms that might lead to ectopic expression of c-fms, the regulation of the exon 2 promoter of the gene, which flanks the transcription start sites in macrophages, was examined. In transient and stable transfections this promoter was as active in non-macrophage tumour cell lines as it was in RAW264 macrophages. Promoter activity in non macrophage lines was serum-dependent and was activated further in lines stably transfected with c-fms. Cis-acting elements required for serum-dependent activity lay outside the 300 bp proximal promoter that was sufficient for maximal activity in RAW264 macrophages, but the c-fms-responsive elements were retained in the proximal promoter. Exon 2 promoter activity was selectively suppressed in non macrophage lines by inclusion of intron 2, which has been implicated in transcription attenuation. Lewis lung carcinoma cells were able to partly bypass this block and expressed c-fms mRNA when grown in limiting serum. The finding that c-fms promoter activity and c-fms mRNA levels are responsive to growth factor signalling pathways provides an insight into mechanisms that may lead to ectopic c-fms expression in tumour cells. PMID- 7478560 TI - Liposome-mediated in vivo E1A gene transfer suppressed dissemination of ovarian cancer cells that overexpress HER-2/neu. AB - The HER-2/neu proto-oncogene is frequently amplified or overexpressed in many different types of human cancers, a phenomenon that has been shown to correlate with shorter survival time and lower survival rate in ovarian cancer patients. We previously reported that increased HER-2/neu expression led to more severe malignancy and increased metastatic potential in animal models and that the adenovirus 5 E1A gene repressed HER-2/neu gene expression at transcriptional level and was able to suppress tumor growth when stably transfected into human ovarian cancer SKOV-3 cells which overexpress HER-2/neu. To investigate whether the E1A gene may be used as a therapeutic agent for HER-2/neu-overexpressing human cancers in living hosts, we first developed tumor-bearing mice by injecting SKOV-3 cells that overexpress HER-2/neu intraperitonealy into female nu/nu mice. Five days later, we used cationic liposomes to directly deliver the E1A gene into adenocarcinomas that developed in the peritoneal cavity and on the mesentery of the mice that received the SKOV-3 cell injection. We found that liposome-mediated E1A gene transfer significantly inhibited growth and dissemination of ovarian cancer cells that overexpress HER-2/neu in the treated mice; about 70% of these mice survived at least 365 days, whereas all the control mice that did not receive the gene therapy developed severe tumor symptoms and died within 160 days. The results suggest that liposome-mediated E1A gene transfer may serve as an effective therapy for human ovarian cancers that overexpress HER-2/neu by directly targeting the HER-2/neu oncogene. PMID- 7478561 TI - Bcl-XL protects cancer cells from p53-mediated apoptosis. AB - Oncogenesis is a process resulting from genetic events which cause loss of growth control or inhibition of appropriate cell death. The Bcl-XL protein is a recently discovered member of the bcl-2 family which has been shown to protect cells from some forms of programmed cell death, but has not yet been implicated in the genesis of human carcinomas. In this report we explore the role of Bcl-XL overexpression in protecting cancer cells from p53-mediated apoptosis. Increased levels of Bcl-XL were found in a subset of primary human breast carcinomas, as well as in the breast cancer line, T47D. T47D cells were then transfected with a temperature-sensitive mutant of the tumor suppressor p53 (p53ts). Although many tumor cell lines undergo apoptosis when p53 is expressed, the T47D transfectants remained viable at temperatures permitting wild-type p53 phenotype. This suggested that endogenous Bcl-XL could protect cancer cells from p53-mediated apoptosis. To test this hypothesis, murine erythroleukemia cells were transfected with bcl-XL and p53ts. While cell lines expressing p53 alone rapidly died, those cells co-expressing Bcl-XL survived. These results demonstrate that Bcl-XL is capable of protecting cells from p53-mediated apoptosis, and suggest a possible mechanism by which tumors expressing Bcl-XL are able to partly overcome the tumor suppressor functions of p53. PMID- 7478562 TI - Somatic mutations of the WAF1/CIP1 gene in primary prostate cancer. AB - The WAF1/CIP1 gene, a potential tumor suppressor gene, has recently been cloned and identified as a p53 mediator and an inhibitor for G1 cyclin-dependent kinases (CDKs). We undertook this study to investigate the possible role of the WAF1/CIP1 gene in human prostatic carcinoma. Matched normal and cancer tissues from 18 patients with prostate cancer were screened for WAF1/CIP1 mutation by nested reverse transcription-polymerase chain reaction/single strand conformational polymorphism (RT-PCR/SSCP) and DNA sequencing. Shifted bands from three tumor, but not the matched normal specimens, were observed. Subsequent direct DNA sequencing of the PCR fragments identified four sequence alterations including a cytosine (C) to adenine (A) transversion and a guanine (G) to A transition and two A insertions. Our results demonstrated that mutations of the WAF1/CIP1 gene occur and may be important during the pathogenesis of human prostate cancer. This is the first report of WAF1/CIP1 mutation in a primary human cancer. PMID- 7478564 TI - Changes in expression of members of the fos and jun families and myc network during terminal differentiation of human keratinocytes. AB - Normal human epidermal keratinocytes are induced to undergo terminal differentiation when disaggregated and placed in suspension: commitment occurs at about 5 h and overt differentiation by 24 h. In contrast, the squamous cell carcinoma line SCC12B2 does not initiate terminal differentiation until 75 h in suspension and the ndk strain of keratinocytes undergoes growth arrest but does not differentiate at all. In order to identify genes that may regulate terminal differentiation we have examined mRNA levels of members of the fos and jun gene families and myc gene network in suspension cultures of normal keratinocytes, SCC12B2 and ndk. The major changes observed were an up-regulation of c-Fos and Fra-1 and a decrease in c-Myc at the time of commitment, followed by an increase in Mad, Mxi1, Fra-2 and JunB expression at the onset of differentiation. The sequence of events shows a striking similarity to that which occurs during myeloid differentiation and suggests a role for AP-1 complexes and Myc and Mad complexes in regulating keratinocyte differentiation. PMID- 7478563 TI - Abnormalities in the CDKN2 (p16INK4/MTS-1) gene in human melanoma cells: relevance to tumor growth and metastasis. AB - The inhibitor of cyclin-dependent kinase 4, CDKN2 (also known as p16INK4 or MTS 1, multiple tumor suppressor gene 1), has been mapped to 9p21. The gene has been shown to be deleted or mutated in high frequency in human melanoma cell lines and familial melanoma patients, suggesting that it could be a melanoma suppressor gene. How these observations are related to tumorigenicity and metastasis of human melanoma is not clear however. To test the role of CDKN2 in human melanoma metastasis, 14 human melanoma cell lines with different metastatic abilities in nude mice were analysed for possible abnormalities in the CDKN2 gene. Homozygous deletions that resulted in a lack of gene expression were found in six of 14 cell lines tested. SSCP-direct sequencing revealed point mutations in three other cell lines. One cell line displayed CC to TT transitions which constitute a hallmark of ultraviolet-induced DNA damage. Overall, abnormalities in the CDKN2 gene were found in nine of 14 (64%) cell lines tested. Homozygous deletion and lack of gene expression were found in several low tumorigenic and nonmetastatic melanoma lines, whereas other metastatic cells did not exhibit abnormalities in the CDKN2 gene. These data suggest that the absence of normal CDKN2 does not confer growth advantage to melanoma cells in vivo and that the production of metastasis by human melanoma cells can occur in the absence of CDKN2 gene abnormalities. PMID- 7478565 TI - Abrogation of p53-induced cell cycle arrest by c-Myc: evidence for an inhibitor of p21WAF1/CIP1/SDI1. AB - The tumor-suppressor p53 inhibits cell cycle progression by direct transactivation of the p21WAF1/CIP1/SDI1 gene, which encodes a universal inhibitor of cyclin dependent kinases (cdk). The proto-oncogene product c-Myc induces cell cycle progression and, in the absence of survival factors, apoptosis. However, a direct link between the cell cycle machinery and c-Myc has not yet been established. We show that c-Myc has not yet been established. We show that c-Myc abrogates a p53-induced G1-arrest without elevating the expression of cdks or cyclins involved in the G1/S-transition. Instead, the results suggest that c-Myc interferes with the inhibitory action of p21 on cdk/cyclin-complexes by inducing a heat-labile inhibitor of p21. The inactivation of p21 and related cdk-inhibitors may explain several of the oncogenic actions of c-Myc, including the induction of proliferation, immortalisation and the inhibition of differentiation. Modulation of cdk activity by the induction of an inhibitor of cdk-inhibitors represents a novel mechanism of cell cycle regulation in mammalian cells. PMID- 7478566 TI - MAP kinase phosphorylation of mSos1 promotes dissociation of mSos1-Shc and mSos1 EGF receptor complexes. AB - The mouse protein mSos1 has a central Ras guanine nucleotide exchange domain, and a long proline-rich C-terminal tail which contains several potential binding sites for the SH3 domains of the adaptor protein, Grb2. In fibroblasts, growth factor stimulation results in the recruitment of Grb2-mSos1 into complexes with activated receptors and cytoplasmic phosphoproteins such as Shc, which are apparently involved in Ras activation, and subsequently to an increase in mSos1 phosphorylation on serine and threonine. The catalytic and C-terminal domains of mSos1 contain several potential sites for phosphorylation by mitogen-activated protein kinases. In vitro, purified p42/p44 MAP-kinase selectively phosphorylated the C-terminal tail of mSos1. Comparative tryptic phosphopeptide mapping of mSos1 phosphorylated in vitro by MAP kinase and of mSos1 immunoprecipitated from EGF stimulated cells, revealed several phosphopeptides in common. These common phosphorylation sites have been mapped to a region encompassing the first three proline (pro)-rich motifs in the tail of mSos1. Furthermore, a region of mSos1 containing the first two pro-rich motifs could associate with MBP kinase activity in vitro. Phosphorylation of mSos1 did not affect binding of Grb2 to mSos1, but appeared to decrease binding of the mSos1-Grb2 complex to Shc and the EGF receptor. These findings suggest a potential inhibitory role for MAP-kinase in attenuating nucleotide exchange on Ras, by uncoupling mSos1 from membrane-bound receptor complexes that lead to Ras activation. PMID- 7478567 TI - The role of the Ataxia telangiectasia gene in the p53, WAF1/CIP1(p21)- and GADD45 mediated response to DNA damage produced by ionising radiation. AB - The inducible response of the tumour suppressor gene p53 has been examined following exposure to DNA-damaging agents in Ataxia telangiectasia (AT) cell lines, an autosomal recessive disorder with multiple clinical and biological abnormalities including sensitivity to ionising radiation. The p53 induction was significantly delayed and reduced in the 8 AT cell lines examined over the 6 h following irradiation with no dose response in p53 induction being observed compared to control cells. The increase of WAF1/CIP1(p21) and GADD45 mRNA, two genes transcriptionally activated by p53, was also reduced in the AT cell lines after such treatment. In contrast, the increase in p53 protein, WAF1/CIP1(p21) and GADD45 mRNA expression following exposure to the alkylating agent methylmethane sulphonate (25 and 100 micrograms ml-1) was similar in both cell types. No alterations in the expression of EBNA-5, an EBV-encoded nuclear antigen which has been shown to bind p53 or mutations in the p53 gene (exons 4 to 8) were found in the AT cell lines studied. The AT gene product would thus appear to be involved upstream of p53, GADD45 and WAF1/CIP1 (p21) in the signalling of the presence of strand breaks produced by ionising radiation, with this defect in response contributing to the high cancer risk and radiosensitivity observed in this disorder. PMID- 7478568 TI - A new member of the DP family, DP-3, with distinct protein products suggests a regulatory role for alternative splicing in the cell cycle transcription factor DRTF1/E2F. AB - Integrating cell cycle progression with transcription provides an important level of control during proliferation. The cellular transcription factor DRTF1/E2F is implicated in this integration process by virtue of its physical interaction and control by key regulators of proliferation, such as retinoblastoma protein, cyclins and cyclin-dependent kinases and regulation of target genes required for cell cycle progression. Generic DRTF1/E2F DNA binding activity arises when a member of two distinct families of proteins, DP and E2F, interact as DP/E2F heterodimers. Here, we report the isolation and characterisation of a new member of the murine DP family, called DP-3 (also referred to as human DP-2). In contrast to previously characterised members of the DP and E2F families, processing of DP-3 RNA provides an important level of control by generating at least four distinct DP-3 proteins, of which three have been isolated, called alpha, beta and gamma. Processing events, which we show are both tissue- and cell restricted, can occur either in the 5' region of DP-3 RNA and determine whether translation begins at one or two potential intiating codons, or within the coding sequence, producing variations in internal domains of the DP-3 proteins. The DP-3 proteins studied can co-operate with E2F-1 in DNA binding activity and trans activation of E2F site-dependent transcription. This analysis of DP-3, which has uncovered a hitherto unexpected and surprising level of complexity, documents a new member of the DP family and novel levels of control which may influence the activity DRTF1/E2F and hence cell cycle progression. PMID- 7478569 TI - Mutagenic analysis of functional domains of the mos proto-oncogene and identification of the sites important for MAPK activation and DNA binding. AB - We constructed in-frame deletion/replacement mutations in the Xenopus mos proto oncogene that lie within conserved Mos-specific codons, but outside of the regions that are conserved among the src kinase family of genes. All gene products were assayed in vitro for kinase activity and in vivo for their ability to induce oocyte maturation, embryonic cleavage arrest and cellular transformation. Most mutations in Mos eliminated both kinase and biological activity. However, a mutation in Mos that removed two basic amino acid residues (R94 and K97) downstream from the lysine at the ATP binding site (K90) markedly enhanced autophosphorylation activity. Moreover, this mutant displayed markedly reduced biological activity, lacked transforming activity, and failed to activate mitogen activated protein kinase (MAPK). A second mutant Mos product, lacking amino acids R45-A54, displayed a five-fold increase in cellular transforming activity. This Mos mutant specifically localized to the cytoplasm; in contrast to wild-type (wt) Mos that localized to both the nucleus and the cytoplasm. These data indicate that Mos transforming activity is mediated via signalling exerted in the cytoplasm, presumably through MAPK, and that nuclear localization of the oncogene product interferes with transforming activity. We also show that amino acids R45-A54 are important for Mos DNA binding activity. PMID- 7478570 TI - Development of cellular resistance to pp60v-src kinase-induced cell death. AB - The v-src gene of Rous sarcoma virus (RSV) encodes pp60v-src, a tyrosine kinase that can initiate cellular transformation. High levels of v-src gene expression can either be cytotoxic or the cause of altered expression of cellular genes. Examination of cytotoxic thresholds is difficult because cells expressing high levels of a cytotoxic oncogene will die. To evaluate quantitatively the cytotoxicity of pp60v-src on growth, we amplified two different v-src genes, under the control of the human hsp70B heat shock promoter to establish cell clones with varying copy numbers of the heat-inducible v-src gene. The viability of cell lines over a prolonged period of time depended on the particular src gene, the expression of v-src mRNA, synthesis of the pp60v-src protein and, most importantly, the tyrosine kinase activity of the pp60v-src protein. We found a relatively sharp threshold in v-src-encoded tyrosine kinase activity above which cell viability rapidly declines. However, over time, tyrosine kinase activity was exponentially suppressed at about a 10-fold higher rate than pp60v-src protein during passage. Our results indicate that homeostasis of tyrosine phosphorylation is important for cell viability, that perturbation of this balance results in cell mortality, and that cells can evolve to accommodate overexpression of oncogene by downregulating the level of tyrosine kinase activity. PMID- 7478571 TI - Tyrosine phosphorylation of murine Crkl. AB - The SH2/SH3 adaptor protein Crkl is abnormally phosphorylated on tyrosine by the Bcr/Abl protein in leukemic cells from patients with Philadelphia-chromosome (Ph)positive leukemia. However, the state of tyrosine-phosphorylation of crkl in normal tissues is unknown. In the current study, we identified mouse crkl by cDNA cloning and examined expression levels and tyrosine-phosphorylation of the mouse crkl protein during embryogenesis and in adult tissues. Tyrosine-phosphorylation of crkl was prominent during early development, but decreased at later embryonic stages and in newborn mice. Expression of both crkl and the related crk was ubiquitous in the adult. However, crkl differed considerably from crk in relative tissue distribution, and was more abundant in hematopoietic tissues. With exception of the lung, crkl was mostly present in a non-tyrosine phosphorylated form. Consistent with our previous findings in human patients, murine crkl was phosphorylated on tyrosine in leukemic tissues of BCR/ABL transgenic animals, but was non-tyrosine phosphorylated in normal mouse bone marrow. We conclude that this crkl tyrosine-phosphorylation by Bcr/Abl in hematopoietic cells is clearly aberrant and is consistently linked to the development of leukemia. Identification of proteins interacting with tyrosine-phosphorylated crkl in the leukemic cells of BCR/ABL transgenic mice should reveal members of signal transduction pathways activated in Ph-positive leukemia. PMID- 7478573 TI - Direction of p21ras-generated signals towards cell growth or apoptosis is determined by protein kinase C and Bcl-2. AB - Normal and activated Ras proteins are known to act as signal transducers, mediating mitogenic responses. Interactions of p21ras and protein kinase C (PKC) are required in a number of mitogenic or activation signaling pathways. The constitutive expression of activated v-Haras in Jurkat cells, a human T lymphoblastoid cell line, renders the cells susceptible to apoptosis during transient down-regulation or inhibition of PKC. Similarly, the expression of v-Ki ras in murine fibroblasts induces apoptosis during suppression of PKC activity. This Ras-specific cell death is dependent upon suppression of cellular PKC activity, and can be blocked by the survival-promoting bcl-2 gene product. In vivo phosphorylation studies indicate that Bcl-2 is a phosphoprotein, and the phosphorylation state of Bcl-2 is modulated in the setting of activated p21Ha-ras in response to inhibition of PKC. These findings suggest an interactive regulation of growth or apoptosis in cells which involves at least three molecules: p21ras, PKC and Bcl-2. PMID- 7478572 TI - Isolation and characterization of a chicken homolog of the E2F-1 transcription factor. AB - In higher eukaryotes, the E2F-1 transcription factor is an essential and limiting component of cell cycle progression in late G1. E2F-1 heterodimerizes with members of the DP gene family and the resulting heterodimer regulates the expression of several proto-oncogenes and the genetic machinery of DNA replication. Cell cycle regulation of E2F activity is mediated through its association with the tumor suppressor Rb gene product. To examine the evolutionary conservation of the E2F-1 protein sequence and its developmental expression pattern we have isolated and sequenced the chick E2F-1 gene (chE2F-1) cDNA. The chicken protein is 34 amino acids (a.a) shorter than its human counterpart (403/437 a.a.) but has extremely well conserved bHLH and pRb binding domains, with respectively 94% and 83% identity. The position of the leucine zipper is also strictly conserved thereby accounting for ability of E2F-1 to form heterodimers with human and chicken DP-1. E2F-1 expression was analysed in synchronized cells as well as in embryonic or newborn chick tissues and appears to be closely correlated to the cell proliferation rate. In situ hybridization studies have shown very high expression levels in the neuroretina during the early stages of embryonic development when active neuroblast division occurs. In contrast, a sharp down-regulation is observed when cells become postmitotic. Overexpression of the chE2F-1 protein leads to oncogenic transformation only when a truncated version of the transgene lacking the pRb binding domain is used; the full length protein either has no effect or may be deleterious for cell survival. PMID- 7478574 TI - v-Myc is invariably required to sustain rapid proliferation of infected cells but in stable cell lines becomes dispensable for other traits of the transformed phenotype. AB - The v-myc-containing retrovirus MC29 induces neoplastic transformation of avian embryo cells. To determine which traits of the transformed phenotype are directly controlled by v-Myc, we engineered a conditional MC29 mutant (GRIM) expressing v Myc as a fusion protein with the glucocorticoid receptor and the retroviral Gag polyprotein. Only in the presence of glucocorticoids such as dexamethasone is GRIM capable of transforming embryo cells, from which six stable GRIM-lines have been derived. Although their survival in culture no longer requires functional v Myc, hormone deprivation causes all six GRIM clones as well as acutely infected fibroblast cultures to either withdraw from cell cycle completely or to grow much more slowly and to much lower densities. However, removal of dexamethasone does not allow GRIM-transformed mass cultures and most of the clones to revert to normal shapes or to reconstruct actin cables. Furthermore, most clones do not require the hormone sustain anchorage-independent growth. We propose that certain secondary events have let the GRIM-clones sustain immortality, transformed morphology, and anchorage-independent growth independently of v-Myc. None of these events, however, has obliterated the requirement for v-Myc in cell division control. We thus conclude that enhanced proliferation is the primary effect of v Myc expression. PMID- 7478575 TI - An E2F-dependent late-serum-response promoter in a gene that controls glycolysis. AB - The F-type mRNA of 6-phosphofructo-2-kinase/fructose-2,6-bisphosphatase is found in proliferating, but not in quiescent, cells. This bifunctional enzyme catalyses the synthesis and degradation of fructose-2,6-bisphosphate, a potent stimulator of glycolysis. F-type mRNA concentration decreased upon differentiation of rat rhabdomyosarcoma cells; it increased in Rat-1 fibroblasts stimulated to proliferate by serum, by epidermal growth factor, or by the v-src oncogene product. This increase resulted, at least in part, from a stimulation of F promoter activity. The stimulation occurred at the G1/S transition of the cell cycle. It depended on a binding site for the oncogenic transcription factor E2F located in the first exon of F-type mRNA. This effect was inhibited by agents that increase cAMP concentration. The data provide the first evidence that a gene involved in the control of glycolysis can be regulated by a late-serum-response promoter in an E2F-dependent way. PMID- 7478576 TI - Analysis of protein-protein interactions involved in the activation of the Shc/Grb-2 pathway by the ErbB-2 kinase. AB - In murine fibroblasts activation of the Shc/Grb-2 pathway by the ErbB-2 kinase involves tyrosine phosphorylation of Shc products and the formation of Shc/ErbB 2, Shc/Grb-2 and Grb-2/ErbB-2 complexes. Tyr 1139 of ErbB-2 bound to the Grb-2 SH2 domain in vitro as well as in intact cells. Tyr 1221 and 1248 are binding sites of gp185ErbB-2 for Shc SH2 domain in vitro whereas Tyr 1196 and 1248 are major binding sites of ErbB-2 for Shc PTB domain. Inhibition of Shc/ErbB-2 complex formation in intact cells was obtained by simultaneous mutational inactivation of Shc SH2 and Shc PTB binding sites of gp185ErbB-2. Shc/ErbB-2 complexes are formed upon activation of the ErbB-2 kinase and tyrosine phosphorylation of Shc proteins; they are located in both cytosol and cellular membranes. ErbB-2 activation induces also translocation of Grb-2 from cytosol to membranes. This network of protein-protein interactions may reflect the ability of the Shc/Grb-2 pathway to act as a molecular switch controlling different cellular functions regulated by RTK activation. In fact the Ras GDP exchanger mSOS was recruited in Grb-2/ErbB-2 complexes; furthermore besides mSOS, other polypeptides present in either cytosolic or membrane preparations were able to complex in vitro with Grb-2 SH3 domains. PMID- 7478577 TI - Activation of the EGF receptor by insertional mutations in its juxtamembrane regions. AB - Ligand dependent activation of receptor tyrosine kinases is mediated by an allosteric dimerization process that is responsible for the stimulation of protein tyrosine kinase activity and receptor autophosphorylation. In order to gain further insight into the processes which control this process, we have generated EGF receptor mutants that contain inserts of 20-40 amino acids in their juxtamembrane regions, on each side of the receptor's single transmembrane domain. An EGF receptor mutant with an insertion on the cytoplasmic side of the transmembrane domain exhibited typical EGF binding characteristics, ligand dependent tyrosine autophosphorylation, as well as ligand-induced DNA synthesis. However, an EGF receptor mutant with an insertion on both sides of the transmembrane domain was found to be constitutively activated. This mutant also exhibited dramatically reduced EGF binding, but dimerized and had enhanced tyrosine kinase activity even in the absence of ligand. Moreover, NIH3T3 cells expressing this mutant receptor formed colonies in soft agar in the absence of EGF. This represents a novel example of a constitutively activated receptor, and provides further support for receptor dimerization as a mechanism for activation of EGFR and other receptor tyrosine kinases. PMID- 7478578 TI - Evidence for different signalling pathways of PKC zeta and ras-p21 in Xenopus oocytes. AB - Considerable effort has been devoted to identifying critical steps in mitogenic signal transduction pathways. Recently, the atypical PKC zeta isoform has attracted great interest since it has been reported to induce GVBD in Xenopus oocytes and transformation of NIH3T3 fibroblasts, two processes closely linked with the regulation of cell division. Furthermore, PKC zeta has been proposed as an essential effector for ras-p21 function and therefore may be an essential component of the signalling pathway(s) activated by mitogens. In this study we have analysed the responses induced in Xenopus oocytes after microinjection of purified recombinant PKC zeta protein. Microinjection of PKC zeta induced the early activation of MPF which precedes GVBD and also induced the activation of MAP kinase and S6 kinase II. The activation of MPF, MAP kinase and S6 kinase II by PKC zeta was sensitive to cycloheximide, while induction of GVBD was independent of protein synthesis. These results indicate that PKC zeta induces the activation of at least two pathways, only one of them leading to the activation of MAP kinase. By contrast, neither the induction of GVBD nor the activation of MPF, MAPK and S6 kinase II induced by the ras-p21 protein were dependent on protein synthesis. Thus, the comparison of these responses suggests that PKC zeta most likely does not mediate the ras-induced signal transduction pathway in Xenopus laevis oocytes. PMID- 7478579 TI - PU.1 (Spi-1) autoregulates its expression in myeloid cells. AB - PU.1 (Spi-1), a member of the Ets transcription factor family, is predominantly expressed in myeloid (granulocytes, monocytes and macrophages) and B cells. PU.1 is upregulated early during commitment of multipotential progenitors to the myeloid lineages and inhibition of PU.1 function in human CD34+ progenitors prior to this upregulation blocks myeloid colony formation. Since PU.1 expression appears to play a role in hematopoietic development, we characterized the PU.1 promoter. Here we report that the murine PU.1 promoter, as well as the human promoter, demonstrate tissue-specific reporter gene expression in myeloid cell lines but not in T cells and HeLa (non-hematopoietic cells) cells. Deletion analysis of the PU.1 promoter indicates that tissue-specific functional elements are encoded in the -61 to -39 bp and -7 to +34 bp regions. The first region contains a functional octamer (Oct) site at -54 bp and an Sp1 site at -39 bp. The second contains a binding site at +20 bp for both PU.1 itself and the related ets family member Spi-B. In vivo footprinting assays demonstrate that a hypersensitive band was detected at the PU.1 site in myeloid cells but not in HeLa. A mutation of the PU.1 site which abolished PU.1 binding caused a significant decrease in promoter activity. Mutation of the Oct and/or Sp1 site results in a lesser decrease of promoter activity in myeloid cells. Co transfection of PU.1 or Spi-B in cells lacking PU.1 and Spi-B specifically transactivated a minimal promoter containing the PU.1 binding site, indicating that PU.1 can activate its own promoter elements in an autoregulatory loop. Positive autoregulation of the PU.1 promoter may play an important role in the function of PU.1 in myeloid cells. PMID- 7478581 TI - Upregulation of vascular endothelial growth factor (VEGF) and downregulation of placenta growth factor (PlGF) associated with malignancy in human thyroid tumors and cell lines. AB - Vascular endothelial growth factor (VEGF) is a potent mitogen for endothelial cells in vitro, promotes neoangiogenesis in vivo and increases the permeability of the vascular endothelium. VEGF overexpression occurs in several cultured tumor cell lines and in certain human malignancies. Placenta growth factor (PlGF) is a recently identified growth factor for endothelial cells (EC); PlGF strongly potentiates both the proliferative and the permeabilization effects exerted by VEGF on the vascular endothelium. To uncover the molecular mechanisms underlying neoangiogenesis in human thyroid tumors, we have analysed VEGF and PlGF expression in a panel of thyroid carcinoma cell lines with different tumorigenic potential as well as in human primary thyroid tumors. We show that a high tumorigenic potential is associated with an elevated VEGF expression in human thyroid tumor cell lines. Furthermore, VEGF overexpression occurs in 5/5 highly malignant anaplastic carcinomas. Papillary and follicular carcinomas express intermediate levels of VEGF mRNA. In contrast, PlGF expression is severely down regulated in the majority of thyroid tumor cell lines and in tumors. Furthermore, we show that both the VEGF receptors, FLT-1 and flk/KDR, are expressed in endothelial cells that line tumor-embedded microvascular vessels, suggesting that VEGF but not PlGF, contributes to thyroid tumor development. PMID- 7478580 TI - The protein product of the c-cbl oncogene rapidly complexes with the EGF receptor and is tyrosine phosphorylated following EGF stimulation. AB - The cbl oncogene was first identified as part of a transforming retrovirus which arose in a mouse pre-B cell lymphoma. Its protein product, p120cbl, is cytoplasmic and has several distinctive domains including a highly basic region, a RING finger motif and a large proline-rich domain. A mutation to cbl in the 70Z/3 pre-B cell lymphoma produces an oncogenic protein which exhibits a marked enhancement of tyrosine phosphorylation. Parallel studies have demonstrated that p120cbl is a substrate of protein tyrosine kinases activated by engagement of the T cell antigen receptor and that cbl is phosphorylated by oncogenic forms of the Abl tyrosine kinase. A genetic analysis of the Caenorhabditis elegans cbl homologue, sli-1, demonstrates that sli-1 negatively regulates the LET-23 tyrosine kinase receptor. Here we show that p120cbl is rapidly phosphorylated on tyrosine residues following EGF stimulation and that it forms an inducible complex with the receptor. Our results also show that the oncogenic 70Z/3 form of cbl has enhanced binding to the EGF receptor and that peptides spanning the proline-rich region bind a range SH3 domains. These findings are consistent with a conserved role for cbl/sli-1 proteins in mammals and nematodes. PMID- 7478582 TI - Evidence for different modes of action of cyclin-dependent kinase inhibitors: p15 and p16 bind to kinases, p21 and p27 bind to cyclins. AB - A number of low molecular weight proteins have recently been identified that specifically inhibit the function of cyclin-dependent protein kinases in mammalian cells. These fall into two distinct families based on primary sequence comparisons and probable modes of action. Using a simple in vitro binding assay, we show that p21CDKN1 and the related p27KIP1 can efficiently interact with cyclins D1, D2, D3, E and A, and to a lesser extent with cyclin B. By generating a deleted form of cyclin D1 that binds to p21 and p27 but not to Cdks, we confirm that these interaction do not depend on stoichiometric amounts of the relevant kinase subunit. Moreover, p21 and p27 do not detectably associated with kinase subunits unless a cyclin is present. This is in sharp contrast to the properties of p16CDKN2 and p15MTS2/INK4b which bind to Cdk4 and Cdk6 in the absence of any cyclin. These data suggest that p21 and p27 act as broad spectrum regulators of cyclin dependent kinase function by participating in ternary complexes whereas the p16 family specifically interfere with the formation of cyclin D-dependent kinase complexes. PMID- 7478583 TI - Identification of an antisense WT1 promoter in intron 1: implications for WT1 gene regulation. AB - Expression of the Wilms' tumour suppressor gene, WT1, is tightly regulated during nephrogenesis, and loss of function of its gene product correlates with malignancy. By using luciferase reporter gene constructs containing DNA sequences from the first intron of the WT1 gene, we have identified an antisense WT1 promoter operational in the opposite direction relative to the 5' promoter. Transcription directed by the promoter is negatively regulated by upstream elements, but is activated by expression of WT1. This effect of WT1 is reciprocal to that observed on the 5' promoter, suggesting that antisense promoter activity is involved in WT1 gene regulation. By mimicking expression of the transcript regulated by the antisense promoter, we demonstrate that cellular levels of WT1 can be effectively down-regulated by antisense mRNA complementary to sequences in the first exon of WT1. PMID- 7478584 TI - Characterization of the signal transduction pathway mediating gamma ray-induced inhibition of DNA synthesis in human cells: indirect evidence for involvement of calmodulin but not protein kinase C nor p53. AB - Cultured cells from patients inheriting the rare cancer-prone and radiotherapy sensitive disorder ataxia-telangiectasia (A-T) exhibit anomalies in cell cycle control and protein kinase C (PKC)-mediated upregulation of p53 protein following exposure to ionizing radiation. It remains unclear, however, as to whether this irregularity in a p53-dependent signal transduction pathway controlling the G1/S checkpoint is causally linked to the most consistent molecular hallmark of A-T namely, marked attenuation in the inhibition of replicative DNA synthesis at early times (< or = 2 h) after irradiation [radioresistant DNA synthesis (RDS)]. We report here that treatment of normal human fibroblast strains with inhibitors of calmodulin (CaM) (i.e. W7 and W13) and CaM-dependent protein kinases II and IV (i.e. KN62) prior to radiation exposure elicits an 'A-T-like' RDS phenotype, whereas treatment with PKC inhibitors (e.g. staurosporine) does not produce this response. Moreover, at 1 h post-gamma irradiation A-T fibroblasts undergo normal induction of p53 protein while exhibiting the RDS trait. At later times (e.g. 4 h) following irradiation, however, these A-T cells contain abnormally low levels of p53 protein, as do their lymphoblastoid cell line counterparts during the entire post-gamma ray incubation period. On the other hand, human cells which either lack the p53 gene completely (i.e. HL60 leukemia cells) or harbor a germline mutation in the gene (i.e. Li-Fraumeni syndrome cells) shut down their DNA replication machinery normally upon sustaining radiation damage. We thus conclude that the transitory delay in DNA synthesis routinely experienced by human cells in the face of radiation injury is mediated through a CaM-dependent regulatory cascade which involves neither PKC nor p53 protein. Accordingly, A-T cells appear to be malfunctional in at least two distinct radiation-responsive signalling pathways, one regulating the G1/S checkpoint and governed by p53 and PKC and another controlling passage through S phase and requiring CaM. PMID- 7478585 TI - Regulation of the Ras signaling pathway by GTPase-activating protein in PC12 cells. AB - We have investigated the role of Ras GTPase-activating protein (GAP) in NGF induced neuronal differentiation by overexpressing both wild-type and membrane targeted GAP in PC12 cells. Extension of neurites in response to NGF was completely blocked in cells expressing the highest level of membrane-targeted GAP and significantly inhibited in cells expressing either wild-type GAP or lower levels of membrane-targeted GAP. Overexpression of membrane-targeted GAP similarly inhibited induction of differentiation by src, but not by ras or raf oncogenes, indicating that GAP inhibits differentiation of PC12 cells by downregulating Ras function. GAP overexpression also inhibited stimulation of mitogen-activated protein (MAP) kinase and induction of immediate-early genes in response to NGF. In cells expressing wild-type GAP or lower levels of membrane targeted GAP, the initial activation of MAP kinase and immediate-early gene expression were only partially inhibited. However, GAP expression in these cells resulted in substantial inhibition of sustained MAP kinase activity following NGF treatment, consistent with the inhibition of neurite extension in these cell lines. These results indicate that GAP acts as a negative regulation, rather than an effector, of Ras signaling in PC12 cells. PMID- 7478586 TI - Characterization of transforming growth factor-beta 1 induced apoptosis in normal human B cells and lymphoma B cell lines. AB - Transforming growth factor beta 1 (TGF beta 1) has been shown to inhibit growth stimulation in normal human B cells as well as in Epstein Barr virus (EBV) negative Burkitt's lymphoma (BL) cell lines. The mechanisms for this potent growth inhibition are not completely defined. Here we show that a number of EBV negative lymphoma B cell lines (BL-41, Ramos and CAPA-2), when exposed in vitro to TGF beta 1, undergo apoptosis. Maximum apoptosis was observed at 48 h following TGF beta 1 treatment, with no apparent effect on the expression of c myc and bcl-2 proteins. Similar induction of apoptosis was observed when these lymphoma cell lines were treated with aphidicolin, a DNA synthesis inhibitor. In contrast, various preparations (14 out of 17) of normal human tonsilar B cells showed no significant apoptosis, although both TGF beta 1 and aphidicolin inhibited anti-mu/IL-4 induced DNA synthesis in all preparations. Furthermore, another TGF beta 1 sensitive EBV-negative BL cell line, CA46, exhibited no apoptosis in response to TGF beta 1 and aphidicolin, corroborating the findings in normal human B cells. Taken together, these data support the hypothesis that exposure to TGF beta 1, which results in cell cycle arrest and DNA synthesis inhibition, may not be obligatory or sufficient for the induction of apoptosis. Rather, induction of apoptosis or lack of it may be intrinsically determined by an interplay between extracellular and intracellular regulators of cellular growth. PMID- 7478587 TI - Cyclic AMP signaling is required for function of the N-terminal and CR1 domains of adenovirus E1A in Saccharomyces cerevisiae. AB - We have constructed yeast vectors in which derivatives of the adenovirus E1A gene are expressed from the GAL1 promoter. Cells expressing E1A289 grow poorly and accumulate cells with a 1C DNA content. Using a series of E1A deletion mutants, we have identified three regions within the E1A protein that are necessary for the G1 growth phenotype; each deletion partially relieves the growth defect. These deletions span residues 4-25, 38-60 and 140-186, which fall within the N terminal, CR1 and CR3 domains of E1A respectively. Expression of the first 82 residues of E1A, spanning just the N-terminal and CR1 domains, strongly inhibits yeast cell growth in G1 showing that these domains can function independently of other domains of E1A. Using this strong growth inhibition, we isolated a yeast mutant in the net1 gene that conferred resistance to the expression of E1A1-82. The mutant was insensitive to expression of both E1A1-82 and full length E1A, but remained sensitive to the toxicity caused by over-expression of a Gal4p-VP16 fusion. Finally, we found that the function of E1A in yeast depends on the cyclic AMP signaling pathway, providing a striking parallel with the action of E1A at the c-fos promoter in mammalian cells. These results suggest that a genetic analysis of the yeast model system will provide relevant new insights into mechanisms of gene regulation by E1A proteins. PMID- 7478588 TI - Enforced expression of full length c-Myb leads to density-dependent transformation of murine haemopoietic cells. AB - The oncogenic activation of c-myb has been associated with structural alterations to the Myb protein. Although such alterations can increase the ability of Myb to transform haemopoietic cells, it has been unresolved whether over-expression of wild type (WT) c-Myb can lead to transformation. We show here that infection with a retrovirus that expresses WT i.e. full length c-Myb leads to transformation of primary haemopoietic cells (as indicated by clonogenic assays). The transformed cells are similar to those obtained with carboxyl-truncated (CT) c-Myb in that they show phenotypic and morphological characteristics of early myeloid cells and remain dependent on exogenous growth factors. Cells expressing WTMyb form lower numbers of colonies on average and have a greater tendency to spontaneously differentiate than those expressing truncated c-Myb. Additionally, our results show that transformation by both forms of Myb is dependent on the density at which the infected cells are cultured, and that low levels of transformation can be increased by addition of conditioned medium from myb transformed cells grown at high density. This implies that transformation can be enhanced by the effects of an autocrine growth factor. Moreover, the production of, or sensitivity to, such a factor may be influenced by Myb itself, since CT Myb-infected cells cultured at low densities show higher levels of transformation than WT Myb infected cells. PMID- 7478589 TI - Evidence for cell-specific differences in transformation by N-, H- and K-ras. AB - Although Ras plays a fundamental role in cellular proliferation, differentiation and transformation, clear functional differences between the three major Ras proteins (N-, H- and K-Ras) have not as yet been demonstrated. In this study, chimeric constructs were used to compare directly transformation by N-, H- and K ras oncogenes. In Rat-2 and NIH3T3 fibroblasts, transformation assays (anchorage independence, focus-formation and growth in 1% FCS) showed that H12-Ras was more transforming than N12-Ras or K12-Ras. By contrast, in the human multipotent haemopoietic cell line, TF-1, N12-Ras exhibited greater biological activity. Northern blotting and protein analyses indicated that these findings were not the result of differences in expression or stability of p21Ras. Using further H-ras/N ras chimeric constructs, we found that the greater transforming activity of H12 Ras in fibroblasts was not due to the hypervariable-CAAX region, but rather to unique sequences between amino acids 84 and 143. These data demonstrate cell specific differences in the intrinsic transforming potential of N-ras, H-ras and K-ras oncogenes. PMID- 7478590 TI - Association of pp60c-src with biliary glycoprotein (CD66a), an adhesion molecule of the carcinoembryonic antigen family downregulated in colorectal carcinomas. AB - CD66a, also known as 'biliary glycoprotein (BGP)', is the human homologue of a cell adhesion molecule (CAM) of the rat (Cell-CAM). CD66a, which belongs to the carcinoembryonic antigen family and the immunoglobulin superfamily, is expressed in cells of myeloid and epithelial origin. The cytoplasmic domain of the major isoform of CD66a (CD66acyt) contains two tyrosine residues in amino acid motifs potentially interacting with protein tyrosine kinases of the Src family. Here we provide evidence that CD66a is associated with pp60c-src. From membrane fractions of granulocytes and the colonic cell line HT29, phosphokinase activity was co immunoprecipitated with CD66a when monoclonal CD66 antibodies or an antiserum against the recombinant cytoplasmic domain of CD66a were used. From the dissociated immunecomplexes, a phosphokinase of M(r) 60,000 was reprecipitated using antibodies against pp60c-src. In vitro, the recombinant cytoplasmic domain was a substrate and binding partner of pp60c-src. Phosphopeptides corresponding to the tyrosine containing amino acid sequences of CD66acyt activated the kinase activity of pp60c-src to a greater extent than a phosphopeptide containing Tyr527 from the SH2-binding regulatory domain of pp60c-src. The down-regulation of CD66a in about 80% of colorectal carcinomas may contribute to a dysregulation of pp60c src in colorectal cancer. PMID- 7478592 TI - The proline-rich region of Vav binds to Grb2 and Grb3-3. AB - Vav has structural features found in signaling proteins and is expressed only in hematopoietic cells. The recent development of mice Vav -/- has confirmed a major role of Vav in early blood cell development. We previously showed that Vav constitutively interacts with glutathione-S-transferase-Grb2. Coimmunoprecipitation experiments supported the idea of a complex formed by Vav Grb2 in vivo. This complex is of potential interest in signaling of hematopoietic cells. In this work we localize the domains of Vav and Grb2 involved in this interaction. By the use of an in vivo genetic approach (the double hybrid system) and through in vitro experiments (glutathione-S-transferase fusion proteins) we furnish evidence that the interaction between Vav and Grb2 involves the C-SH3 domain of Grb2 and the proline-rich region located in the N-SH3 of Vav. Furthermore this was confirmed by the use of both Vav and Sos derived proline rich peptides which blocked the binding. In addition we show that Vav also interacts with Grb3-3, a naturally occurring Grb2 isoform wich lacks functional SH2 domain. PMID- 7478591 TI - The murine BCL6 gene is induced in activated lymphocytes as an immediate early gene. AB - The chromosomal translocation involving 3q27 is often detected in human B-cell lymphomas, especially diffuse lymphomas with a large-cell component. The BCL6 gene has been isolated from the chromosomal breakpoint in these lymphomas. Here we cloned the murine BCL6 (mBCL6) cDNA from the muscle cDNA library using the human BCL6 (hBCL6) cDNA as a probe. The predicted amino acid sequence was 95% identical to that of hBCL6. It contains six repeats of the Kruppel-like zinc finger motif that are completely identical to those of hBCL6, indicating that the BCL6 gene is well conserved between humans and mice. Expression of the mBCL6 gene was ubiquitously detected in adult mouse tissues including lymphatic organs. Furthermore, it was induced in lymphocytes activated with phorbol ester and Ca2+ ionophore within 30 min after stimulation. This induction was not inhibited by treatment of the cells with a protein synthesis inhibitor, cycloheximide. These results suggest that BCL6 plays a role in activated lymphocytes as an immediate early gene. PMID- 7478593 TI - Detailed deletion mapping of chromosome 9q in bladder cancer: evidence for two tumour suppressor loci. AB - Loss of heterozygosity (LOH) at loci on chromosome 9p and/or 9q is the most frequent genetic alteration in transitional cell carcinoma (TCC) of the bladder. However, localisation of the tumour suppressor locus or loci on 9q has been hampered by the relative infrequency of tumours with subchromosomal deletions. We have used 24 microsatellite markers to examine LOH in 70 new cases of TCC of the bladder and upper urinary tract. Forty tumours (57%) showed LOH at one or more loci on 9q and partial deletions were detected in five tumours (7%). Combined data from the five cases with partial deletions place one tumour suppressor locus at 9q34 between D9S61 and D9S66 (an estimated distance of 13-14 cM). This region is frequently deleted in other sporadic tumours and encompasses one of the loci for tuberous sclerosis (TSC1). One tumour contained a distinct deletion between D9S153 and D9S109 (9q13-q31), which encompasses the locus for the familial nevoid basal cell carcinoma syndrome (Gorlin syndrome). This may indicate the presence of another tumour suppressor locus on 9q for TCC. Our findings significantly reduce the regions of 9q within which suppressor genes for TCC may reside. The possible involvement of two deletion targets on 9q in addition to the locus at 9p21 implicated in TCC may explain why LOH at all loci on chromosome 9 is frequent in TCC. PMID- 7478594 TI - Gadd45 is a nuclear cell cycle regulated protein which interacts with p21Cip1. AB - GADD45 was originally identified as a cDNA clone induced by growth arrest and DNA damage. We show that Gadd45 is a nuclear protein, widely expressed in normal tissues, particularly in quiescent cellular populations. Using cell synchronisation methods we show that Gadd45 levels are highest in the G1 phase of the cell cycle, and are greatly reduced during S phase. Immunoprecipitation of Gadd45 from mammalian cells reveals that it is tightly associated with a protein which reacts with antibodies to the cyclin dependent kinase inhibitor p21Cip1. Binding of recombinant Gadd45 protein to overlapping p21Cip1 peptides in ELISA assays and use of the yeast two hybrid assay show that Gadd45 directly interacts with this cell cycle inhibitor. These data suggest that Gadd45 may act in the regulation of the cell cycle. It is postulated that the interactions of Gadd45 with both p21Cip1 and PCNA are important for the modulation of cell cycles, and for the inhibition of DNA replication. PMID- 7478595 TI - Regulation of E2F-1 gene expression by p130 (Rb2) and D-type cyclin kinase activity. AB - Previous experiments have demonstrated that the regulation of E2F-1 transcription factor activity is critical for the maintenance of normal cell proliferation control. Regulation of E2F-1 is accomplished through at least two mechanisms: posttranslational regulation by binding proteins such as Rb and transcriptional regulation of the E2F-1 gene. The E2F-1 gene promoter has recently been isolated to examine this latter aspect of E2F-1 regulation. Preliminary studies demonstrate that the E2F-1 promoter is under E2F-dependent negative control during the cell growth response, being transcriptionally repressed through E2F sites in G0 and early G1. We now demonstrate that the presence of an E2F DNA binding complex containing the Rb-related p130 protein (Rb2) correlates with E2F 1 gene repression and that overexpression of p130 inhibits transcription from the E2F-1 promoter. Moreover, D-type cyclin-dependent kinase activity specifically activates the E2F-1 promoter by relieving E2F-mediated repression but is inhibited by coexpression of the cdk4 and cdk6 inhibitor p16 (CDKN2, MTS1, INK4). Taken together, these findings suggest that E2F-1 gene expression is controlled during cell cycle progression by a regulatory network involving at least one oncogene (cyclin D1) and several potential tumor suppressor genes. PMID- 7478596 TI - A novel Bcl-2 related gene, Bfl-1, is overexpressed in stomach cancer and preferentially expressed in bone marrow. AB - Programmed cell death (apoptosis) is an active process which is genetically encoded and plays an important role in several cellular activities such as embryonic development, deletion of autoreactive T-cells and homeostasis. Several genes regulating apoptosis have been reported, including p53, one of the tumor suppressor genes, c-myc, one of the proto-oncogenes, and various kinds of Bcl-2 related genes. A new cDNA clone which is homologous to Bcl-2, named as Bfl-1 were isolated from a human fetal liver at 22 week of gestation. This clone was identified by computer analysis of random cDNA sequences that were obtained in an effort to expand the expressed sequence tag (EST) databases to be used for human genome analysis. The homology was recognized by 72% amino acid identity to the murine A1 gene, a member of the Bcl-2-related genes. The homology to the BH1 and BH2 domains of Bcl-2 was especially significant, suggesting that Bfl-1 is a new member of the Bcl-2-related genes. Bfl-1 is abundantly expressed in the bone marrow and at a low level in some other tissues. Interestingly, a correlation was noted between the expression level of Bfl-1 gene and the development of stomach cancer in eight sets of clinical samples. It is conceivable that Bfl-1 is involved in the promotion of the cell survival in the stomach cancer development or progression. PMID- 7478597 TI - Tumor induction by v-Jun homodimers in chickens. AB - To study the contribution of v-Jun homodimers to oncogenesis, we constructed artificial v-Jun derivatives in which the natural dimerization domain of v-Jun was replaced by an heterologous homodimerization domain from either the viral EB1 or the yeast GCN4 transcription factor. The resulting v-Jun chimeric proteins, called v-Juneb1 and v-Jungcn4, which can no longer dimerize with Jun or Fos, should only form homodimers in the cell. Helper-independent retroviruses expressing v-Jun, v-Juneb1 and v-Jungcn4 were generated. All three viruses transformed primary cultures of chick embryo cells with the same high efficiency and promoted local tumor growth after subcutaneous injection of infected cells in young animals. In contrast, after intravenous injection of viral suspensions into chick embryos, only the chimeric proteins produced internal tumors that were lethal. These tumors were leiomyosarcomas located within the liver and along the digestive tract. Thus, in vivo, v-Juneb1 and v-Jungcn4 are more potent oncoproteins than v-Jun. These data demonstrate that when forced to accumulate, v Jun homodimers can induce tumors efficiently. They also show that the oncogenic potential of v-Jun can be regulated through the properties of its dimerization domain. PMID- 7478598 TI - The Epstein-Barr virus encoded cytokine viral interleukin-10 enhances transformation of human B lymphocytes. AB - In vitro infection of human B lymphocytes with Epstein-Barr virus (EBV) results in their growth transformation and establishment of immortalised lymphoblastoid cell lines. The virus was recently found to encode a homologue of the pleitropic cytokine interleukin-10 (IL-10), which has wide ranging effects on the immune system. We have investigated the effect of this virally encoded growth factor on the ability of EBV to immortalize B lymphocytes from tonsils and from adult and neonatal blood. Recombinant viral interleukin-10 (vIL-10) was found to increase dramatically the growth transformation of B cells from all three populations infected with either the highly transforming type 1 strain B95-8 or the less efficient type 2 strain BL16. This striking enhancement of transforming ability in the presence of viral IL-10 may be in part due to increased viability of the B cells during infection and decreased levels of interferon-gamma, a cytokine known to inhibit EBV transformation. Thus viral IL-10 influences a number of cell types of the immune system to allow the enhanced outgrowth of EBV transformed cells. PMID- 7478599 TI - c-Myc and Max transregulate the mouse ornithine decarboxylase promoter through interaction with two downstream CACGTG motifs. AB - Ornithine decarboxylase (ODC), the first enzyme in the biosynthesis of polyamines, is essential for the process of cellular proliferation. ODC is a typical delayed early gene, as its mitogenic activation requires ongoing protein synthesis in the stimulated cells. This study provides evidence that the immediate early c-Myc protein is a potential transactivator of the ODC gene. We demonstrate that overexpression of c-Myc results in efficient activation of the ODC promoter, whereas overexpression of Max exerts a repressive effect. Both effects depend on the presence of two evolutionary conserved CACGTG motifs found in the first intron of the ODC gene. Transactivation of the ODC promoter also requires the dimerization of c-Myc with Max. Interestingly, over-expression of USF, a member of the same family of proteins which efficiently binds these two CACGTG motifs, fails to transregulate the ODC promoter. Our data suggest that c Myc and Max are potential transcriptional regulators of the ODC promoter. PMID- 7478600 TI - Moloney murine leukemia virus infection accelerates lymphomagenesis in E mu-bcl-2 transgenic mice. AB - E mu-bcl-2 transgenic mice bearing the bcl-2 proto-oncogene linked to the immunoglobulin enhancer (E mu) sporadically develop B or T cell lymphomas after a long latent period. To identify genes that play important roles in development of lymphoid malignancies, proviral insertional mutagenesis with Moloney murine leukemia virus (MMuLV) was carried out in two lines of transgenic mice expressing the bcl-2 gene primarily in B or T cells. MMuLV infection of non-transgenic mice induced primarily mature T cell lymphomas. By contrast, infection of newborn E mu blc-2 mice with the virus accelerated lymphomagenesis, and nearly all of the mice eventually succumbed to clonal pre-B, B, or mainly immature T cell lymphoma, indicating the active contribution of the bcl-2 gene in lymphomagenesis. Southern blot analysis of tumor DNA from MMuLV-infected transgenic mice revealed a proviral insertion at the c-myc gene in 26% (9/35) of tumors, at the pim-1 gene in 6% (2/35) and at the pim-2 (recently renamed tic-1) gene in 23% (8/35). Some tumors carried two activated oncogenes. No insertion was detected at the bmi-1 gene. These data suggest the usefulness of this transgenic system for analysis of lymphomagenesis involving the activated bcl-2 gene. PMID- 7478601 TI - The physical map of the human RET proto-oncogene. AB - The RET proto-oncogene, a transmembrane tyrosine kinase receptor, is involved in the development of at least five different disease phenotypes. RET is activated through somatic rearrangements in a number of cases of papillary thyroid carcinoma while germ-line point mutations are associated with three inherited cancer syndromes MEN 2A, MEN 2B and FMTC. Moreover, point mutations or heterozygous deletions of RET are found in the dominant form of Hirschsprung disease or congenital colonic aganglionosis. We cloned the entire RET genomic sequence in a contig of cosmids encompassing 150 kb, from the CA repeat sTCL-2 to the region upstream the RET promoter, and established the position of the 20 exons of the RET gene with respect to a detailed restriction map based on eight endonucleases. A new highly polymorphic CA repeat sequence was identified within intron 5 of RET (RET-INT5). Finally the orientation of RET on chromosome 10q11.2 made it possible to orientate three other genes rearranged with RET in papillary thyroid carcinomas, namely H4/D10S170 on 10q21, R1 alpha on 17q23 and RFG2/Ele1 on 10q11.2. PMID- 7478602 TI - Novel members of the eph receptor tyrosine kinase subfamily expressed during Xenopus development. AB - Three cDNAs encoding receptor tyrosine kinases (RTKs) of the eph-subfamily have been identified based on their homology to Pagliaccio (Winning and Sargent, 1994, Mech. Dev. 46:219-229). These have been named TCK, Xelk (Xenopus homologue of elk), and PL7a (pag-like clone 7a). Each of these genes is expressed in a distinctive, tissue specific manner during early development. TCK is expressed in pre-somitic mesoderm, caudal somites, midbrain and cement gland. Xelk is expressed in the brain and spinal cord and in the first and fourth visceral arches. PL7a cDNA is expressed throughout the head and in the tip of the tail. All of the genes are represented in maternal mRNA, and are expressed in adult tissues. The Xelk cDNA encodes a protein which is 94% identical to rat elk and therefore is likely to represent the Xenopus homologue of this gene. TCK and PL7a are less related to previously identified eph-subfamily RTKs. The inability to unambiguously assign TCK and PL7a as Xenopus homologues of any previously identified eph RTK leads us to conclude that these cDNAs represent novel members of this family. PMID- 7478603 TI - Clonal expansion of germline B-lineage acute lymphoblastic leukemia in severe combined immunodeficient mice. AB - CD19+ B lineage acute lymphoblastic leukemias (ALLs) with unrearranged Ig and TCR genes are designated germline B lineage ALLs. We used CDR3 PCR to determine whether pediatric germline B lineage ALLs contain minor subclones with rearranged Ig H V genes. In six of seven cases there were no PCR detectable CDR3 rearrangements. One case with a smear pattern on CDR3 PCR contained multiple unique CDR3 sequences at frequencies of 1-2 per 2,600, suggesting that polyclonal B cells were present at low frequency. To verify that the germline patterns were from leukemic cells and evaluate in vivo subclone differentiation, a germline B lineage ALL with the t(4;11) translocation was propagated in severe combined immunodeficient SCID) mice. The Ig and TCR genes in the leukemic cells recovered from mouse tissues were germline by Southern blot analysis except for single rearrangements that suggested subclone evolution at the Ig H and lambda loci in addition to the germline population. No CDR3 sequences were detected, indicating that the observed Ig H gene rearrangement most likely was a DJ joining. This study suggests that the transformed cell in germline B lineage ALL represents an early pro-B cell with limited tendency to further differentiate. PMID- 7478604 TI - Functional domains of the t(8;21) fusion protein, AML-1/ETO. AB - The AML-1/ETO fusion protein is created by the (8;21) translocation, the second most frequent chromosomal abnormality associated with acute myeloid leukemia. In the fusion protein the AML-1 runt homology domain, which is responsible for DNA binding and CBF beta interaction, is linked to ETO, a gene of unknown function. The primary sequences of the runt homology domain indicates no known DNA binding motifs, but is predicted to contain six beta-strands, two alpha-helices and a nucleotide binding motif. Mutagenesis of AML-1/ETO was performed to delimit the functional domains of the chimeric protein. Most mutations in the runt homology domain that resulted in reduced CBF beta binding also inhibited DNA binding, indicating that the DNA and CBF beta binding sequences are tightly linked. However, these activities were separated by a point mutation of residue 144, within the putative ATP binding motif, which nearly eliminated DNA binding, but did not affect CBF beta binding. Random mutagenesis identified the hydrophobic face of the amphipathic fifth beta-strand, adjacent to the putative ATP binding motif, as critical for both DNA and CBF beta binding. C-terminal deletion mutants of AML-1/ETO indicated that ETO sequences are essential for interference with AML 1B-mediated transcriptional activation, and that residue 540 defines the C terminal boundary of a potential repression domain. Thus, these mutational analyses define the regions of AML-1/ETO which regulate its function and that may be important in promoting leukemia. PMID- 7478605 TI - Dissociation of transcriptional activation and oncogenic transformation by v-Myb. AB - The nuclear oncoprotein v-Myb is a transcriptional activator in both animal cells and the budding yeast Saccharomyces cerevisiae. Previous studies have suggested that an acidic domain of approximately 50 amino acids (amino acids 204-254 of v Myb) is necessary and sufficient for transcriptional activation by v-Myb, c-Myb and GAL4-Myb fusion proteins. However, we find that first, none of the acidic residues within this region is essential for transcriptional activation in either animal cells or yeast. Second, transcriptional activation requires cooperation among multiple domains of v-Myb. In animal cells, transcriptional activation by v Myb requires a central domain (amino acids 234-295), a C-terminal domain (amino acids 295-356), plus either of two more N-terminal domains (amino acids 163-197 or 198-232); in yeast, it requires the central domain plus either the C-terminal domain or a more N-terminal domain (amino acids 163-233). Third, although various subsets of these domains are sufficient for transcriptional activation by v-Myb, all of the domains must be present for transformation of primary hematopoietic cells. These results demonstrate that transcriptional activation by v-Myb is not sufficient for oncogenic transformation. PMID- 7478606 TI - The evolution of WT1 sequence and expression pattern in the vertebrates. AB - WT1 is a Wilms' tumour predisposition gene, encoding a protein with four C terminal Kruppel-type zinc fingers, which is also a major regulator of kidney and gonadal development. To pinpoint key regulatory domains involved in development and evolution of the vertebrate genitourinary system, we have isolated WT1 orthologues from all gnathostome classes. Partial nucleotide sequence from chick, alligator, Xenopus laevis and zebrafish reveals extensive conservation. Both the zinc fingers and the transregulatory domain exhibit a high level of similarity in all the species examined. However, of the two alternatively spliced regions only one, the three amino acid KTS insertion between zinc fingers 3 and 4, is found in species other than mammals. The 17 amino acid insertion at the C-terminal end of the transregulatory domain is present only in mammals. Residues with reported human pathological mutations are also unaltered across species, underlining their structural significance. Studies in chick and alligator reveal that the mammalian intermediate mesoderm expression pattern is conserved in birds and reptiles. A wider role in mesodermal differentiation is suggested by expression in some paraxial and lateral mesoderm derivatives. PMID- 7478607 TI - Identification and functional characterization of the human and murine polo-like kinase (Plk) promoter. AB - The Plk gene encodes a serine/theronine kinase which is located in the nucleus. Northern blot analysis linked Plk expression to the proliferative activity of cells and tissues. To analyse the transcriptional regulation of the Plk gene we have isolated several human genomic clones containing the Plk promoter. RNAse protection assays revealed three major transcription start sites within a 40 bp region centered around the 5' end of the known human cDNA and 6 minor Cap sites. A genomic fragment of 2.3 kb located 5' to the translation start sites drives the expression of the CAT-reporter in transient transfections in human (EPLC, HeLa) and mouse (NIH3T3, 32D) cell lines in an orientation dependent fashion. The 2.3 kb genomic fragment contains a CCAAT motif located 30-70 bp upstream of the Cap sites and two overlapping Sp1 sites 20 bp further upstream. Additional sequence motif homologues to binding sites of known transcription factors could be identified. In addition to the human Plk promoter, the mouse Plk promoter was isolated. The sequence alignment of the human and murine promoter revealed three regions with extensive sequence homology within a region of 300 bp immediately upstream of the Cap sites. A fourth region of homology encompassing 90 bp about 2.1 kb 5' of the Cap sites was identified as well. Deletion of various regions within the 2.3 kb promoter fragment identified several domains involved in the regulation of the human Plk promoter. The 300 bp region immediately 5' of the Cap sites which is highly conserved between mouse and man is essential for promoter activity. 3' deletions including the CCAAT site abolished promoter activity. Growing 5' deletions within the core region of the promoter reduces transcriptional activity. Furthermore, using deletion clones we identified regions 5' of the core region which enhance or silence the transcriptional activity of the core promoter. PMID- 7478608 TI - Activation of Src family kinases in Neu-induced mammary tumors correlates with their association with distinct sets of tyrosine phosphorylated proteins in vivo. AB - Overexpression and amplification of the erbB-2 (neu) is thought to play a major role in mammary cancer. Although studies suggest that Neu is directly involved in the genesis of mammary tumors, the molecular mechanism by which Neu induces tumors is not well understood. Recently, we have demonstrated that the activity of c-Src tyrosine kinase is elevated in Neu-induced mammary tumors and this elevated activity correlates with its capacity to physically associate with Neu. To explore whether other members of the c-Src family are activated in these mammary tumors, we measured the in vitro kinase activity of the c-Yes and Fyn kinases in protein extracts derived from mammary tumor tissue and morphological normal adjacent tissue. These analyses revealed that c-Yes kinase activity was elevated in Neu-induced tumors by comparison to the adjacent tissue. By contrast, no significant activation of the Fyn kinase was noted in these tumors. Activation of c-Yes tyrosine kinase correlated with the capacity of c-Yes to associate with Neu in vivo in lysates derived from primary tumor samples. Studies with Rat.2 fibroblasts overexpressing activated Neu revealed that c-Src requires the presence of tyrosine phosphorylated Neu for its ability to interact with Neu in vivo. Moreover, analyses using radiolabeled c-Yes SH2 fusion protein revealed that this interaction is likely occurring in a direct fashion. Although both c Src and c-Yes kinase associate with Neu in vivo, a tyrosine phosphorylated protein of 89 kd (p89) was found associated with c-Src but not with c-Yes in cell lysates derived from mammary epithelial cells transformed by either Neu or PyV middle T antigen. Furthermore, this tyrosine phosphorylated protein was not detected in c-Src complexes derived from fibroblasts transformed by either Neu or PyV middle T. These observations suggest that p89 associates with c-Src only in mammary epithelial cells and not in fibroblasts. PMID- 7478609 TI - Allelic loss of distal chromosome 4 in mouse lung tumors localize a putative tumor suppressor gene to a region homologous with human chromosome 1p36. AB - Loss of heterozygosity (LOH) of distal chromosome 4 was studied to localize a putative tumor suppressor gene in mouse lung neoplasia. Previous studies have implicated this region and its homologue on human chromosome 1p36-34 as sites of potential tumor suppressor genes. One hundred adenocarcinomas and 38 adenomas of the mouse lung were examined for LOH using nine simple sequence length polymorphism markers in a polymerase chain reaction-based approach. Forty-four adenocarcinomas displayed allele loss, all of which included losses at marker D4MIT54, which defined a critical region of 3 centiMorgans as the likely location of a putative tumor suppressor gene. In 58% of the tumors displaying LOH, all markers used in this study incurred allele loss. In contrast, retention of heterozygosity was observed at all markers tested in each of the adenomas studied, which suggests that the inactivation of this tumor suppressor gene participates in mouse lung tumor progression. PMID- 7478610 TI - The role of ovarian hormones, age and mammary gland development in polyomavirus mammary tumorigenesis. AB - Polyomavirus infection of adolescent athymic female mice causes a high incidence of mammary adenocarcinomas. We have examined the role of ovarian hormones, age and mammary gland developmental stage at infection on subsequent tumor induction, viral replication and gene expression. Ovariectomy (OVX) of adolescent mice 1 week before infection decreased mammary tumor incidence and number, and significantly increased tumor incidence and number, and significantly increased tumor latency. Reduction in tumorigenesis was observed to a lesser degree if mice were OVX at the time of or after infection, indicating that ovarian hormones are mainly required for tumor initiation. Tumor incidence was also reduced with increasing age; OVX prior to infection at older ages drastically reduced tumor development. Treatment of OVX adult mice with estrogen + progesterone for 1-3 weeks prior to infection was unable to restore tumorigenesis to the level observed in intact mice. Thus, in contrast to adolescent mice, the continued presence of ovarian hormones after infection was required for maximal tumorigenesis in adult mice. The decreased tumorigenesis observed in older animals is not likely due to increased differentiation since late pregnant mice with well differentiated mammary glands remained highly susceptible to tumorigenesis. At 10 days post infection, the levels of viral genomes were moderately high and similar in all experimental groups. Early viral protein and middle T-associated kinase levels were undetectable in infected tissues in all experimental conditions. However, high levels were found in tumors, perhaps reflecting a high dosage requirement for oncogenesis. PMID- 7478611 TI - Allelotype of neuroblastoma. AB - Although relatively high incidence of loss of heterozygosity (LOH) on chromosomes 1p, 11q, and 14q have been reported in neuroblastoma, it is still unclear whether or not LOH occurs specifically on these chromosome arms in neuroblastoma since only a few chromosomal arms have been examined for LOH in previous studies. Therefore, we screened 81 cases of tumors for LOH on all 22 autosomes and chromosome X using 35 restriction fragment length polymorphism markers and eight microsatellite markers. High incidence of LOH (> 20%) was observed on six chromosome arms; 1p (26%), 2q (30%), 9p (36%), 11q (24%), 14q (22%), and 18q (31%). Frequencies of LOH on other chromosome arms were less than 13%. Patients with 9p LOH in the tumors showed statistically significant association with advanced stage of the disease and poor prognosis (P = 0.037 and P = 0.003, respectively) independently from N-myc amplification, while LOH on other chromosomes did not show association with stage, prognosis, and N-myc amplification. Thus, besides LOH on chromosomes 1p, 11q, and 14q, LOH on chromosomes 2q, 9p, and 18q also occurs relatively frequently in neuroblastoma, indicating the involvement of multiple tumor suppressor genes in the development of neuroblastoma. It is possible that there is a novel tumor suppressor gene on chromosome 9p which is involved in the progression of neuroblastoma. PMID- 7478612 TI - Highly-expressed p100/p52 (NFKB2) sequesters other NF-kappa B-related proteins in the cytoplasm of human breast cancer cells. AB - Several observations have suggested that NF-kappa B transcription factors could be involved in carcinogenesis. To investigate the possibility that members of the NF-kappa B family participate in the molecular control of the transformed phenotype, we examined the expression of these proteins in human breast cancer cell lines as well as in primary tumors. Western Immunoblots demonstrated high expression of the p52 precursor p100 (NFKB2) in several breast cancer cell lines while human mammary epithelial cells express this protein only faintly. Eighteen primary breast tumors out of 24 displayed significant expression of the p100/p52 protein. In MDA-MB-435 cells, overexpressed p100 and p52 are predominantly cytoplasmic and coimmunoprecipitation experiments demonstrated that p100 sequesters the heterodimer p50/p65 in the cytoplasm. We demonstrate that most p65 protein is complexed with p100 in these cells while it is complexed predominantly with I kappa B-alpha in cell lines expressing less p100. Our data strengthen the hypothesis that NF-kappa B could be involved in carcinogenesis and suggest that the p100/p52 NF-kappa B subunit could play a role in the development of human breast cancers, possibly by sequestering other NF-kappa B-related proteins in the cytoplasm. PMID- 7478613 TI - Mechanism of inactivation of CDKN2 and MTS2 in non-small cell lung cancer and association with advanced stage. AB - The CDKN2 tumor suppressor gene encodes an inhibitor of type D cyclin dependent kinases. CDKN2 is homozygously deleted in approximately 25% of nonsmall cell lung cancer (NSCLC) cell lines and these deletions are associated with advanced stage cancer. Conflicting reports of the frequency of CDKN2 alterations in NSCLC tumors prompted us to examine the relationship of these alterations and those of the related gene, MTS2, with patient stage and site of cancer. One hundred twenty five NSCLC samples (71 cell lines and 54 tumors) were examined by PCR-SSCP. Twenty of 71 (28%) tumor cell lines had homozygous deletions, and six (8%) had point mutations compared to 4 (7%) with point mutations among 54 tumor samples. All mutations were observed in tumors or cell lines from patients with stage III or IV disease. Two patients with no mutations in their primary tumor had a CDKN2 point mutation detected in a metastatic tumor. Point mutations were G:C to T:A transversion on the coding strand in five of 10 and resulted in nonsense mutations in seven of 10. Undetectable CDKN2 mRNA, in the absence of detectable genetic alteration, was noted in a similar fraction of cell lines derived from patients with stage I or II disease [two of seven (29%)] and stage III or IV disease [15 of 49 (31%)]. Homozygous deletion of MTS2 was found in 17 of 20 cell lines with CDKN2 deletions; no point mutations of MTS2 were identified by SSCP in the 125 samples. Thus, CDKN2 is a frequent target of genetic alterations at 9p21 in NSCLC. Both deletions and point mutations of CDKN2 are closely associated with tumor dissemination. PMID- 7478614 TI - The anti-proliferative effect of vitamin D3 analogues is not mediated by inhibition of the AP-1 pathway, but may be related to promoter selectivity. AB - The hormone 1,25-dihydroxyvitamin D3 (VD) is able to induce cellular differentiation and to inhibit cellular proliferation, which provides it with an interesting therapeutic potential in cancer. However, side effects of VD on homeostasis (eg hypercalcemia) had made the need for the development of VD analogues with low calcemic effect. On the human breast cancer cell line MCF-7 we obtained with the VD analogue EB1089 an about 100-fold higher anti-proliferative effect than with VD. We found that this difference in biological activity is neither related to increased functional affinity to the VD receptor nor to repression of AP-1 activity. The physiologically most prominent complex of the VD receptor is a heterodimer with the retinoid X receptor that binds VD response elements formed two hexameric core binding motifs being arranged either as direct repeats spaced by 3 nucleotides (DR3s) or as inverted palindromes spaced by 9 nucleotides (IP9s). We observed that EB1089 stimulates transcriptional activation from IP9-type elements at clearly lower concentrations than from DR3-type elements. It is possible that IP9-type response elements play an important role in or contribute to the control of cell proliferation, so that promoter selectivity may explain the high anti-proliferative effect of EB1089. PMID- 7478615 TI - The viral oncoproteins Ad5 E1A, HPV16 E7 and SV40 TAg bind a common region of the TBP-associated factor-110. AB - A function shared by the adenovirus E1A, papillomavirus E7 and SV40 TAg oncoproteins is their ability to interfere with normal cell growth by interacting with members of the retinoblastoma protein family. In this study, we show that each of these oncoproteins can also bind to the 921 amino acid TBP-associated factor-110 (TAF-110). The significance of the binding is underscored by the observation that each oncoprotein binds to the same 77 amino acid carboxyl region of TAF-110. In the case of E1A and TAg, this finding is consistent with their abilities to stimulate transcription initiation, in part, through their known interactions with TBP. While it is not clear whether E7 can also activate promoters through protein:protein interactions with components of the transcription initiation complex, our demonstration that E7 can bind to TAF-110, as well as TBP, suggests that E7 may modulate the expression of specific promoters which could contribute to the pathogenesis of human papillomavirus. PMID- 7478616 TI - Loss of transcriptional attenuation in N-myc is associated with progression towards a more malignant phenotype. AB - The modulation of RNA polymerase II processivity through the untranslated N-myc first exon represents an important mechanism governing N-myc mRNA levels during normal development. In this study, we employed the rat embryo fibroblast (REF) cooperation assay as a functional means to (i) map N-myc first exon sequences involved in the regulation of N-myc gene expression and specifically their contribution to controlling a block to transcriptional elongation and (ii) determine whether this transcriptional control mechanism plays a role in governing the oncogenic activity of N-myc. Transfection of N-myc expression constructs harboring various deletions within the untranslated first exon revealed that a region encoding a potential stem-loop structure followed by a thymine stretch (stem-loop/T region) was required for efficient transcriptional attenuation. The increase in transcriptional read-through associated with all deletions involving the stem-loop/T region also resulted in a more aggressive malignant phenotype as evidenced by a significant enhancement in the subcloning efficiency of N-myc/ras transformed foci. Most significantly, when cell lines generated from overexpression of the intact N-myc expression construct were selected for anchorage-independent growth, a strong block to transcriptional elongation was completely eliminated in all cases examined. Since the subcloning efficiency of transformed foci and the capacity of permanently established cell lines for anchorage-independent growth are direct correlates of more advanced stages of malignant transformation, our findings suggest that loss of transcriptional attenuation represents an important genetic event in the progression of N-myc-induced cellular transformation. PMID- 7478617 TI - An Inr-containing sequence flanking the TATA box of the human c-sis (PDGF-B) proto-oncogene promoter functions in cis as a co-activator for its intronic enhancer. AB - High-level activity of the human PDGF-B promoter in choriocarcinoma cell lines depends upon an atypical, intronic enhancer-like element which does not function with heterologous promoters tested. An extensive series of mutant PDGF-B promoter driven constructs identified a sequence flanking the TATA box which is required specifically for enhancer-mediated transcription in human choriocarcinoma cell lines. This element, which we here term an enhancer-dependent cis co-activator (EDC) contains an Inr (initiator) consensus sequence upstream of the TATA box which is required, but not sufficient for its function. Requirement for the EDC is cell type-specific, since it was dispensable for enhancer-mediated transcription in a human breast cancer cell line. Although it lies within the region defined, the TATA box itself is not required for EDC function, or for basal promoter function which may derive from a second Inr-like sequence situated at the transcriptional start site. These observations indicate that interactions between some promoter and enhancer elements may be more complex than that generally described for 'classical' enhancer systems and may suggest an additional function for the initiator motif. PMID- 7478618 TI - Diminished capacity for p53 in mediating a radiation-induced G1 arrest in established human tumor cell lines. AB - It has been reported that the p53 gene mediates an ionizing radiation-induced G1 arrest in mammalian cells. To further characterize this important phenomenon, a panel of seven human diploid fibroblast cell strains and 14 human tumor cell lines from a variety of sources with both wild-type and mutant p53 status were assayed for their susceptibility to G1 arrest after gamma-ray irradiation by a continuous labeling [3H]thymidine incorporation technique. An irreversible G1 block involving 20-70% of the cell population was observed in diploid fibroblasts irradiated with 4 Gy. The block was abolished by transfection with the Human Papilloma Virus E6 gene and in an ataxia telangiectasia (AT) cell line, indicating a role for the AT and p53 genes respectively in this process. In contrast to wild-type normal fibroblast cell strains, the G1-block in all tumor cell lines was significantly reduced, irrespective of their p53 status. None of the nine human tumor cell lines with mutant p53 genes showed a significant G1 block following irradiation with 4 Gy. Among the five tumor cell lines expressing wild-type p53, two showed no apparent G1-block. The remaining three showed a G1 block involving only 8-15% of the cell population, a block much smaller in magnitude than that seen in diploid fibroblasts. Finally, a diploid fibroblast cell strain and a tumor cell line, both showing a normal p53 and p21/WAF1 expression pattern, were examined for pRb phosphorylation before and after irradiation. The diploid fibroblast cell strain showed a significant G1-arrest and a clear inhibition of pRb phosphorylation by irradiation whereas the tumor cells showed no G1-arrest and no inhibition of pRb phosphorylation. These results suggest that (1) multiple genetic factors may modulate the occurrence and magnitude of the G1-arrest induced by exposure to ionizing radiation, (2) the capacity for p53 to mediate a radiation-induced G1 arrest is significantly reduced in tumor cells, (3) the disruption of G1-block modulating factor(s) other than p53 may be an important step in carcinogenesis. PMID- 7478619 TI - Expression of cyclin D1 mRNA is not upregulated by Myc in rat fibroblasts. AB - Conflicting results have been published regarding the regulation of cyclin D1 mRNA in rat fibroblasts expressing a hormone-regulated Myc protein, MycER. We confirm that activation of MycER with oestrogen rapidly induces cyclin D1 mRNA, even in the presence of cycloheximide. However, we show that this is an artefact resulting from an oestrogen-activated transcriptional activation domain in the oestrogen receptor part of the MycER chimaera. First, addition of 4-hydroxy tamoxifen (4OHT), which does not activate this domain, allows association of MycER with Max and induces cell proliferation in serum-starved Rat-1-MycER cells without affecting cyclin D1 mRNA levels or the activity of D1 promoter-luciferase constructs. Second, Rat-1 cells expressing a mutant MycER with a hormone-binding domain that still binds 4OHT but no longer binds oestrogen, are driven into the cell cycle in response to 4OHT but fail to up-regulate cyclin D1 mRNA. Finally, Rat-1 cells in which wild-type human c-Myc expression can be induced, also progress into the cell cycle without increased D1 mRNA expression. PMID- 7478620 TI - Overexpression of p21WAF1/CIP1 induces growth arrest, giant cell formation and apoptosis in human breast carcinoma cell lines. AB - The p21WAF1/CIP1 protein was shown to be a critical downstream effector of p53 and a potent inhibitor of cyclin-dependent kinases. We investigated the effects of exogenous p21WAF1/CIP1 in two different human breast carcinoma (HBC) cell lines MCF-7 and T47D. p21WAF1/CIP1 transfected cells formed significantly reduced number of drug-resistant colonies than their mock-transfected counterparts. The majority of the p21WAF1/CIP1 transfected MCF-7 cells acquired 50-100-fold increase in their sizes and persisted as single giant cells for several days without cell division, while the remainder underwent nuclear division (multiple nuclei) but were unable to complete cytokinesis, and finally all succumbed to apoptosis. Only few p21WAF1/CIP1 transfected T47D cells displayed increase in their sizes while a number of them formed small sized drug-resistant colonies which diminished in size due to cell death. The cells in these shrinking colonies exhibited characteristic signs of apoptosis such as chromatin condensation and fragmented nuclei while their membrane integrity was preserved. Thus in HBC cell lines the growth suppression due to enforced overexpression of p21WAF1/CIP1 is associated with giant cell formation and apoptosis. PMID- 7478621 TI - Activating mutations of the TSH receptor in differentiated thyroid carcinomas. AB - A series of 14 thyroid carcinomas, characterized for their basal adenyl cyclase activity (ACA), was examined for the presence of activating point mutations in the TSH receptor (TSHR) gene. Sequencing of the carboxyl-part of this gene revealed the presence of a somatic and heterozygotic point mutation in codon 623 in three out of six tumors showing a constitutively enhanced ACA and a poor response to TSH stimulation. The mutation determines the substitution of a serine for an alanine in the third intracellular loop of the receptor, in a region critical for signal transduction. One tumor bearing a TSHR mutation presented also a N-ras point mutation. Both mutations were detected also in a lung metastasis of this tumor. Our data represent the first report of alterations in the TSHR gene in thyroid malign neoplasia. TSHR mutations may indeed participate, as well as the G alpha s protein (gsp oncogene), in the oncogenesis of some differentiated thyroid carcinomas presenting increased basal levels of cAMP and a poor response to TSH. PMID- 7478622 TI - Interaction between murine germline mutations in p53 and APC predisposes to pancreatic neoplasia but not to increased intestinal malignancy. AB - Murine strains which bear constitutive inactivating mutations of either the APC or the p53 tumor suppressor genes are characterised by spontaneous tumors. APC mutated (Min) mice develop large and small bowel adenomas, a small proportion of which, in time, become malignant. p53 deficient mice develop predominantly lymphoma and sarcoma. By interbreeding these strains we have shown that there is co-operativity between these mutations, leading to a shift in phenotype. Most notably, this was characterised by a range of abnormalities of the exocrine pancreas in 83% of animals heterozygous for the APC mutation and constitutively null for functional p53. Dysplasia and preneoplastic foci were seen in 61% of these animals and pancreatic acinar cell adenocarcinoma in 22%. Analysis of these tumors showed them to have lost the remaining wild-type copy of APC. Similar loss of APC was not associated with the development of other extra-intestinal tumors. Surprisingly, given the proposed role for loss of function mutations of the p53 gene in the development of human colorectal cancer, we have found no evidence for either an increase in the rate of adenoma formation in APC +/-, p53 -/- animals, or an increased rate of progression to malignancy compared with APC +/- p53 +/+ mice. These findings highlight striking tissue-specific differences in the tumor suppressor effects of p53. PMID- 7478624 TI - Measurement of letter acuity in preschool children. PMID- 7478623 TI - Bik, a novel death-inducing protein shares a distinct sequence motif with Bcl-2 family proteins and interacts with viral and cellular survival-promoting proteins. AB - The survival-promoting activity of the Bcl-2 family of proteins appears to be modulated by interactions between various cellular proteins. We have identified a novel cellular protein, Bik, that interacts with the cellular survival-promoting proteins, Bcl-2 and Bcl-xL, as well as the viral survival-promoting proteins, Epstein Barr virus-BHRF1 and adenovirus E1B-19 kDa. In transient transfection assays, Bik promotes cell death in a manner similar to the death-promoting members of the Bcl-2 family, Bax and Bak. This death-promoting activity of Bik can be suppressed by coexpression of Bcl-2, Bcl-XL, EBV-BHRF1 and E1B-19 kDa proteins suggesting that Bik may be a common target for both cellular and viral anti-apoptotic proteins. While Bik does not show overt homology to the BH1 and BH2 conserved domains characteristic of the Bcl-2 family, it does share a 9 amino acid domain (BH3) with Bax and Bak which may be a critical determinant for the death-promoting activity of these proteins. PMID- 7478625 TI - Using visual thresholds to establish low vision performance. AB - Table 6 gives an example of a similar task (N10 print of 75% contrast) but with different requirements of fluency showing how much extra visual performance (acuity threshold, contrast threshold and magnification) is needed to achieve fluent reading. After the clinical assessment of visual function, it should be possible to categorize clients into three different groups on the basis of their threshold visual performance, each needing a different approach depending on their goals and needs as shown in Table 7. These are: Group I: Limited or no survival reading potential. Group II: Survival reading potential but limited likelihood of fluency. Group III: Clear survival reading potential with fluent reading possible if motivated. The author believes that it is always best to start with what the client has to read in order to maintain their independence, rather than what they wish to read for leisure, education or employment. Therefore, it is more successful to concentrate initially on spot/survival reading even if the client seems to have potential for regaining fluency. It should, however, be made clear to the client that this is the approach to be adopted. It is futile for the practitioner, as well as frustrating and demoralizing to the client, to press ahead with complex devices or wasteful trial and error sessions if, at an early stage, it is clear that the client will never return to fluent reading. Instead, it is far better to begin the move to vision substitution (e.g. audio-tapes) for leisure reading at an earlier stage. The overriding influence in low vision management is clearly the motivation of the client.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7478626 TI - Treatment of myopia with the excimer laser--is it really the bottom line? PMID- 7478627 TI - Are we ready to use genetic testing to diagnose predisposition to cancer? PMID- 7478628 TI - Psychosocial oncology clinical nurse specialist enhances staff and patient well being. Interview by Mary Beth Tombes. PMID- 7478629 TI - 1992: engaging a transforming world. PMID- 7478630 TI - Airway obstruction. The problem and its causes. AB - Chronic airway obstruction usually has an insidious onset. It is often difficult to diagnose unless the patient is examined by a physician familiar with the problem and causes. Potential causes can be divided into infectious, inflammatory, traumatic, and neoplastic. Although the precise incidence of each category is unknown, trauma, secondary to previous prolonged intubation for mechanical ventilation, is currently the most common premorbid factor. PMID- 7478631 TI - Anatomy of the larynx, trachea, and bronchi. AB - This article reviews the anatomy of the larynx, trachea, and bronchi. It emphasizes the structure of the laryngeal framework, the innervation and blood supply of the larynx and trachea, the anatomic differences between the pediatric and adult larynx and trachea, and the relational anatomy of structures surrounding the trachea and bronchi. PMID- 7478632 TI - Radiology of the central airways. AB - Diagnostic imaging of the central airways has evolved from being the blind spot in the chest radiograph to involving complex imaging techniques in a region of great pathologic diversity. Just as diagnostic imaging has become more sophisticated, so have the techniques now available for the surgical treatment of these disorders. Close cooperation and consultation between the clinical and diagnostic teams will remain the hallmark of optimal patient care in this sophisticated diagnostic and treatment environment. PMID- 7478633 TI - Flow volume loops in the evaluation of upper airway obstruction. AB - Patients with lesions that cause obstruction of the large airways are often misdiagnosed as having chronic lung disease or reactive airways disease. Close attention to the history and physical examination provides clues to the presence of a laryngeal or tracheal lesion. Obtaining a flow volume loop in the pulmonary function laboratory is a simple and effective method of noninvasively evaluating a patient for the presence of an upper airway obstruction. Fixed lesions cause plateaus in both the inspiratory and expiratory limbs of the flow volume loop. Variable intrathoracic lesions are characterized by expiratory slowing and flattening of the expiratory limb. An important caveat is that these changes may not be present in a patient with coexisting lower airway disease such as COPD or asthma. Variable extrathoracic lesions cause inspiratory slowing and a plateau on the inspiratory limb of the flow volume loop. Finally, the clinician should remember that the quality of the flow volume loop is totally dependent on the patient's effort and cooperation and, thus, that the tracings obtained in the pulmonary function laboratory may not have the classic shapes presented in this review. PMID- 7478634 TI - Video laryngoscopy. AB - New techniques for visualing the upper and part of the lower airway provide for efficient assessment of airway dysfunction. Equipment is available for video recording during the assessment, which can be done either in the caregiver's office/clinic or during stress testing in an exercise facility. Case reports that illustrate the diagnostic use of video laryngoscopy as well as adjunct to therapy are presented in this article. PMID- 7478635 TI - Operative evaluation of airway obstruction. AB - Endoscopic evaluation is a crucial component in the assessment of chronic airway obstruction. Acute airway obstruction is a potentially devastating complication of this invasive procedure. This article discusses a systematic approach to endoscopically assessing the airway safely and obtaining the information necessary for long-term management of the chonically obstructed airway. PMID- 7478636 TI - An overview of treatment options for lower airway obstruction. AB - Effectively treating airway stenosis is a challenging problem for the otolaryngologist-head and neck surgeon. A number of options are available. The most effective management requires an approach that is flexible and individualized for each patient. This article outlines the available options and highlights the indications, techniques, results, and potential complications of each. PMID- 7478637 TI - Laser treatment of lower airway stenosis. AB - This article reviews the various laser techniques used in the endoscopic treatment of lower airway stenosis. The tests of lasers and laser settings used are presented along with the various success rates for the particular techniques. Also presented in this article are the criteria for differentiating those patients who are likely to have a good response to endoscopic techniques versus those patients who should have open techniques performed. PMID- 7478638 TI - Open surgical techniques for laryngotracheal stenosis. AB - Adult subglottic and proximal tracheal stenoses are some of the most crippling complications of endotracheal intubation or other laryngotracheal trauma. Subglottic stenoses are a part of the broader category of laryngotracheal stenoses that comprises any lesion from supraglottic larynx to trachea. Patients with laryngotracheal or tracheal stenosis may present with stridor, shortness of breath, or exercise intolerance and may be tracheotomy dependent. The surgeon must have several choices of procedures available to correctly treat the variety of lesions that may occur. This article briefly outlines the pathophysiology of subglottic and tracheal stenosis and discusses in detail the open procedures available. PMID- 7478639 TI - Diagnosis and management of pediatric laryngeal stenosis. AB - Pediatric laryngeal stenosis presents diagnostic and management challenges. A team approach involving multiple pediatric subspecialties is desirable. Management includes conservative endoscopic and aggressive open surgical intervention. Deciding which approach is appropriate presents a diagnostic dilemma. The long-term prognosis for most children is very good, and the successes are emotionally rewarding. PMID- 7478640 TI - Diagnosis and management of pediatric tracheal stenosis. AB - Pediatric tracheal stenosis is a difficult problem that requires very careful preoperative evaluation and classification. Most forms of extrinsic compression are vascular in origin but can be associated with tracheomalacia, complete tracheal rings, and other anomalies of the aerodigestive tract. Optimally, both the vascular malformation and the intrinsic tracheal malformation should be repaired at the same procedure. Long tracheal stenosis, most commonly caused by complete tracheal rings, is best treated by pericardial patch tracheoplasty or cartilage augmentation. Areas of short-segment intrinsic stenosis can sometimes be corrected endoscopically with or without intraluminal stenting, depending on severity. When endoscopic methods fail, resection with end-to-end anastomosis is the procedure of choice for short-segment stenosis. PMID- 7478641 TI - Reinnervation of the allograft larynx in the rat laryngeal transplant model. AB - The rat model for a vascularized laryngeal allograft is duplicated with significant technical modifications. We report the addition of unilateral host-to allograft recurrent laryngeal nerve anastomosis to this model. Long-term survival experiments determine the feasibility of reinnervation studies of the allograft larynx with this new mode. A total of 59 transplants have been performed on histocompatible Munich Wistar rats, 36 with attempted unilateral allograft reinnervation. Because of the initially high operative mortality rates, additions and modifications of the original technique resulting in reproducibly enhanced survival are detailed. Factors critical to the functional study of this model with regard to reinnervation are elucidated. Preliminary data on allograft reinnervation are reported as confirmed by videodocumentation of vocal fold mobility, evoked and spontaneous electromyography, and glycogen-depletion studies. PMID- 7478642 TI - Mitochondrial role in hair cell survival after injury. AB - The role of mitochondrial biogenesis in hair cell survival after injury was evaluated by inhibiting mitochondrial protein synthesis with chloramphenicol and then studying the effects on hair cell survival after exposure to two different types of ototoxins, gentamicin and acoustic trauma. Seven- to 10-day-old chicks were treated with either a single injection of gentamicin (250 mg/kg) or noise (1500 Hz at 120 dB sound pressure level for 14 hours). A subset of the gentamicin and noise-treated animals also received chloramphenicol (1200 mg/kg during a 24 hour period) through a subcutaneous osmotic pump. A control group received chloramphenicol alone (1200 mg/kg during a 24-hour period). All animals were sacrificed after 5 days, and their basilar papillae (cochleas) were prepared for scanning electron microscopy. Hair cell loss was quantified with stereologic techniques. Animals treated with chloramphenicol alone did not have any evidence of hair cell loss. Gentamicin-treated animals had characteristic hair cell loss beginning at the basal tip and tapering out along the inferior edge more distally. The addition of chloramphenicol to gentamicin treatment significantly increased hair cell loss by 30%, extending the area of hair cell loss into the superior hair cell region at the distal boundary of the lesion. Pure-tone noise exposure characteristically produced hair cell loss along the inferior edge and occasionally included hair cells along the most superior edge. Addition of chloramphenicol to noise exposure significantly increased hair cell loss by 80%, with extension of the lesion across the full width of the sensory epithelium and basally. These results demonstrate that mitochondrial biogenesis is involved in cellular responses to injury. They suggest that mitochondrial function may regulate the probability of survival after metabolic challenges to hair cell integrity. PMID- 7478643 TI - Immunohistochemical characterization of nitric oxide synthase activity in squamous cell carcinoma of the head and neck. AB - This study was designed to investigate the presence of nitric oxide in human squamous cell carcinoma of the head and neck. We localized the activity of nitric oxide synthase in these tumors through immunohistochemical analysis using antibodies to L-citrulline (a byproduct of nitric oxide synthase), to inducible nitric oxide synthase, and to constitutive nitric oxide synthase. We found presence of inducible enzyme in squamous cells throughout these tumors, with the highest intensity staining occurring directly around keratin pearls. Our findings suggest that inducible nitric oxide synthase activity is present in squamous cell carcinomas of the head and neck, leading us to conclude that inducible nitric oxide synthase may play a significant role in tumor growth. PMID- 7478644 TI - Signal averaging and waveform analysis of laser Doppler flowmetry monitoring of porcine myocutaneous flaps: I. Acute assessment of flap viability. AB - Postoperative monitoring of microvascular free-tissue transfer is essential to the early identification and correction of vascular compromise. Laser Doppler flowmetry is a noninvasive monitor of capillary bed perfusion. Its current clinical use requires continuous monitoring and trend analysis to detect changes in capillary perfusion. This study investigated the hypothesis that signal averaging of laser Doppler flowmetry output triggered by a fixed point in the cardiac cycle would provide accurate information about the microvascular flow patterns not dependent on trend analysis. These results indicate that averaged waveform analysis allowed for a rapid, objective, and statistically significant distinction between a viable myocutaneous flap and one with vascular compromise in a porcine model. Moreover, this technique allows for distinction between venous and arterial insufficiency. PMID- 7478645 TI - Human papillomavirus types important in progression of inverted papilloma. AB - Recent evidence suggests that human papillomavirus may play a role in the pathogenesis of inverted papilloma, a benign but locally aggressive neoplasm with a high recurrence rate and an association with squamous cell carcinoma. Histologic features of inverted papilloma have not been useful in discriminating lesions at high risk for malignant transformation. We studied archival pathology specimens from 39 patients with inverted papilloma treated at the University of Michigan between 1980 and 1994 using polymerase chain reaction techniques and human papillomavirus L1 and E6 consensus primers. Previously we reported that 63% of these specimens tested positive for human papillomavirus sequences and that presence of human papillomavirus predicted recurrence of inverted papilloma. We used type-specific primer pairs and polymerase chain reaction techniques as well as hybridization with type-specific oligonucleotide probes to determine human papillomavirus type. A significant correlation was observed between the severity of the lesion (dysplasia or carcinoma) and high risk human papillomavirus type (p < 0.01). All 12 benign inverted papilloma specimens that contained human papillomavirus tested positive for human papillomavirus 6 or 11. Of seven inverted papilloma specimens that exhibited dysplasia, five were human papillomavirus positive, three contained human papillomavirus 6, one contained human papillomavirus 11, and one contained human papillomavirus 18. In each of the three specimens that contained inverted papilloma in association with squamous cell carcinoma, the inverted papilloma portion of the specimen tested positive for a single human papillomavirus type: human papillomavirus 6, 11, or 16.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7478646 TI - Transverse cervical artery fasciocutaneous free flap for head and neck reconstruction: initial anatomic and dye studies. AB - METHOD: The bilateral transverse cervical arteries of 16 fresh cadavers were exposed by an infraclavicular midline approach. Each artery was cannulated, and methylene blue dye was infused to delineate the skin territory subserved by the vessel. The two major infusion skin patterns obtained allowed fashioning of a thin fasciocutaneous flap incorporating supraclavicular skin, which was based on the transverse cervical artery; or a larger flap additionally incorporating upper back skin and varying amounts of trapezius muscle, when the artery had a dorsal scapular artery branch. Depending on the skin pattern, either scapula or clavicle could be transferred with the other soft tissues. RESULTS: The skin territory of the transverse cervical artery is caused to vary by the presence or absence of its dorsal scapular artery branch. The supraclavicular portion of the flap is recommended for repair of facial and nasal lesions because of its close match in color and texture to facial skin. Oral lesions can also be reconstructed with this flap because of its pliability. CONCLUSION: The free flap based on the transverse cervical artery pedicle appears to be a useful addition to the armamentarium of flaps for head and neck reconstruction. Clinical use of the flap is ongoing and will be subsequently reported. PMID- 7478647 TI - A mechanism for sympathectomy-induced bone resorption in the middle ear. AB - BACKGROUND: Recent investigations have demonstrated a link between sympathectomy and osteoclast-mediated bone resorption. The exact nature of this link, however, is unknown. We hypothesize that substance P, a potent vasoconstrictive neuropeptide found in peripheral sensory fibers, including those innervating bone, is the mediator of this phenomenon. To test this theory, the effects of substance P on in vitro calcium release from cultured neonatal mouse calvaria were assessed. In addition, an in vivo study was conducted whereby gerbils were injected with capsaicin to eliminate substance P-containing fibers before sympathectomy with 6-hydroxydopamine. If the effects of 6-hydroxydopamine were eliminated by prior administration of capsaicin, the role of sensory nerves in sympathectomy-induced resorption would be strongly implicated. IN VITRO STUDY: Substance P at 10(-8) mol/L was incubated with eight newborn Swiss-Webster mouse hemicalvarial explants and compared with explants incubated in control media alone. The neonatal mice were euthanized at day 3, and their hemicalvaria were preincubated in 2 ml of stock media without treatment for 24 hours at 36.5 degrees C as a stabilization period. After the stabilization period, the stock media were replaced with 2 ml of fresh control media or media containing substance P at 10(-8) mol/L. A similar experiment was performed with the addition of indomethacin at 5 x 10(-7). The explants were then incubated for 72 hours with gassing every 12 hours with a mixture of O2, N2, and CO2. At the end of the 72 hour period, the media were analyzed for calcium content by atomic absorption spectrophotometry and compared by one-way analysis of variance with Bonferroni corrected post hoc tests. IN VIVO STUDY: Forty-eight Mongolian gerbils were placed into four groups: group 1 received intraperitoneal injections of 6 hydroxydopamine at 75 micrograms/gm body weight on days 1, 2, 6, 7, and 8; group 2 received identical injections of hydroxydopamine, but 12 hours after receiving subdermal injections of capsaicin at 50 micrograms/gm body weight; group 3 received only subdermal injections of capsaicin; and group 4 received only saline injections to serve as controls. Seven days after treatment, the animals were euthanized, and the ventral wall of each animal's right bulla was resected and quantified for osteoclast number and surface with a computer-based histomorphometry system. Analysis was then made by one-way analysis of variance with Bonferroni-corrected post hoc tests. RESULTS: The results of the in vitro study revealed that substance P at 10(-8) mol/L (11.05 +/- 3.37 micrograms/ml) induced significant calcium release from cultured neonatal mouse calvaria when compared with control bone incubated in base media alone (0.92 +/- 2.85 micrograms/ml, p < 0.01). The process was completely inhibited by 5.0 x 10(-7) indomethacin. The results of the in vivo study showed 6-hydroxydopamine treatment significantly increased both the osteoclast number (NOc/TL = 3.14 +/- 1.33/mm) and the osteoclast surface (OcS/BS = 16.04% +/- 6.95%) of bone when compared with bone from saline-treated controls (NOc/TL = 1.77 +/- 0.79/mm, p < 0.01; OcS/BS = 8.88% +/- 4.15%, p < 0.01). These 6-hydroxydopamine-induced increases were eliminated, however, in animals pretreated with capsaicin before sympathectomy (NOc/TL = 1.86 +/- 0.68/mm, p > 0.05; OcS/BS = 9.92 +/- 3.73, p > 0.05), whereas treatment with capsaicin alone had no effect when compared with bone from saline treated controls (NOc/TL = 2.02 +/- 0.50/mm, p > 0.05; OcS/BS = 10.28% +/- 2.62%, p > 0.05). Substance P has thus been shown to induce calcium release from membranous bone in vitro, whereas capsaicin, a substance P-specific sensory neurolytic chemical, eliminates the in vivo osteoclast-inductive effects of 6 hydroxydopamine when given 12 hours before treatment. The results indicate that substance P is capable of inducing resorption and that substance P-containing sensory ne PMID- 7478649 TI - Outcome analysis for lip carcinoma. AB - Carcinoma of the lip is a relatively common malignancy of the head and neck region, accounting for approximately one quarter of oral cavity cancers. Although this form of cancer is generally readily curable compared with malignancies at other head and neck sites, regional metastases, local recurrence, and even death from this disease may occasionally occur. A review of 1252 patients who were diagnosed with lip cancer from 1940 to 1987 was undertaken to identify and rank prognostic variables, clarify differential incidences of site predilection between male and female patients, and examine the correlation between tumor site and histopathologic diagnoses. Large tumor size, high tumor grade, the presence of adenopathy, a subsite other than the lower lip, and inadequate surgical margins were found to have a negative impact on determinate survival of patients with lip carcinoma. Twenty-one percent of lip cancers in female patients arose on the upper lip, whereas only 3% of lip cancers in male patients developed on the upper lip. Basal cell carcinomas comprised 13% of upper lip cancers and only less than 1% of lower lip cancers. Recurrence developed in 15.1% of the patients reviewed and was strongly associated with large tumor size and poor differentiation. Local recurrence was associated with a determinate survival of 78%, whereas patients having regional recurrences had a survival rate of 52%. These data support aggressive treatment of lip cancers greater than 3 cm in diameter, high-grade tumors, tumors associated with cervical lymphadenopathy, and upper lip and commissure tumors. Lip cancer in women seems to be slightly more aggressive than in men. PMID- 7478648 TI - Signal transduction mechanisms in substance P-mediated ciliostimulation. AB - Substance P is a neuropeptide released by afferent neurons in the respiratory tract during inflammatory reactions. It produces effects on blood vessels, bronchial smooth muscle, nasal glands, and respiratory cilia. We studied the in vitro effect of substance P on the ciliary beat frequency of human adenoid explants and its mechanism of action. Substance P was added to cultured adenoid at concentrations of 10(-10), 10(-8), 10(-6), and 10(-4) mol/L. Ciliary beat frequency was determined with phase-contrast microscopy and microphotometry. Substance P increased ciliary beat frequency a maximum of 11.9% +/- 3.8% (p < 0.01). Diclofenac (10(-6) mol/L) significantly blocked the ciliostimulatory effects of SP (p < 0.022), indicating that prostaglandin synthesis is an intermediate step in the action of substance P on ciliary beat frequency. The L arginine analogs, NG-nitro-L-arginine methyl ester and NG-monomethyl-L-arginine, inhibit nitric oxide synthesis from L-arginine. L-Arginine analogs (10(-4) to 10( 2) mol/L) inhibited the effect of substance P (p < 0.02 at the higher concentration). This inhibition was reversed by adding L-arginine, demonstrating that nitric oxide production is a required step in substance P-induced ciliostimulation. Substance P stimulates ciliary activity in human nasal mucosa as a result of secondary production and release of endogenous prostaglandins and nitric oxide. It is likely that inflammatory disease processes that stimulate release of substance P and subsequent prostaglandin and nitric oxide production modify mucociliary transport. Pharmacologic modification of substance P and its second messengers may eventually permit regulation of this important defense mechanism and control of neurogenic inflammation. PMID- 7478650 TI - Immunotherapy basics. AB - Immunotherapy is the use of controlled exposure to allergens to produce durable antiinflammatory effects, thus reducing the severity of allergic disorders. Immunotherapy is useful when other methods of allergy therapy are not fully satisfactory and can be effectively combined with rhinologic surgical treatment. Immunotherapy should always be considered as a treatment option for allergy patients and can often be of benefit, provided that appropriate indications and contraindications are observed. Physicians caring for patients with allergies should therefore become familiar with methods of allergy and diagnosis and with the therapeutic potential of immunotherapy. The history of immunotherapy, possible mechanisms, indications, contraindications, testing methods, and initiation of treatment are reviewed. PMID- 7478651 TI - Future immunotherapy: what lies ahead? AB - There is currently great interest in developing improved methods of immunotherapy and new techniques of immune system manipulation to ameliorate allergic diseases. This article reviews current research trends in the immunologic treatment of allergy, including the use of chemically modified allergens, nonparenteral allergen exposure, sustained-release allergen delivery, anti-immunoglobulin E antibodies, gamma-globulin, immune complexes, cytokines, and T-cell-tolerogenic peptides. PMID- 7478652 TI - Large vestibular aqueduct syndrome with massive endolymphatic sacs. PMID- 7478653 TI - Prosthetic management of velopharyngeal insufficiency induced by uvulopalatopharyngoplasty. PMID- 7478655 TI - Vallecular cyst synchronous with laryngomalacia: presentation of two cases. PMID- 7478654 TI - Inflammatory meningiomas of the internal auditory canals. PMID- 7478656 TI - Transoral endoscopic nasopharyngeal surgery. PMID- 7478657 TI - Follicular variant of papillary carcinoma with hyperthyroidism. PMID- 7478658 TI - Toxic shock syndrome in a patient using bilateral silicone nasal splints. PMID- 7478660 TI - Multichannel cochlear implantation in a child with Brachmann-de Lange syndrome. PMID- 7478659 TI - Malignant melanoma of the lacrimal sac. AB - Although malignancies of the lacrimal sac are rare, malignant melanoma of the lacrimal sac is an exceedingly rare occurrence. Patients with abnormalities of the nasolacrimal system may come to otolaryngologists for evaluation. Treatment of lacrimal sac malignancies will necessitate consultation of a head and neck surgeon. The triad of symptoms commonly found in these patients includes epiphora, bloody reflux from the lacrimal punctum, and a medial canthal mass. Early local recurrence occurs frequently. Radical surgical treatment, including wide-field en bloc resection of the entire nasolacrimal system, affords the best protection against local recurrence and provides the best opportunity for enhanced patient survival. Adjunctive therapy in this disorder remains controversial. PMID- 7478661 TI - Tuberculosis: an unusual cause of dysphagia. PMID- 7478662 TI - Congenital lacrimal sac mucoceles with intranasal extension. PMID- 7478664 TI - State society supports rural patient access bill. PMID- 7478663 TI - Severe congenital glottic web. PMID- 7478665 TI - Managing Medicaid: community plan finding success in Lancaster. AB - After more than two years in development, the Lancaster Community Health Plan on May 3 opened its phone lines to Medicaid patients in need of a "medical home." Now, four months later, physician and community response to this new managed care plan is overwhelmingly positive--and snags are few and far between. PMID- 7478666 TI - Physicians should beware of Medicare secondary payer laws. PMID- 7478667 TI - Medical uses of the Internet. AB - Unless you've spent the last couple of years in Antarctica or maybe Ulan Bator, you've been reading a lot about the Internet. All the hype and hoopla may leave you wondering whether the Internet will be useful for you. The answer is somewhere between "probably" and an unqualified "yes," depending on whether your perspective is short term or long term. This article, one of a series on applications of personal computers for physicians, describes some of the resources available on the Internet. PMID- 7478668 TI - Medical students tackle adolescent health. AB - Vincent J. Zarro, MD, PhD, is assistant dean of student affairs and director of the division of addiction medicine in the Department of Medicine at the Medical College of Pennsylvania and Hahnemann University School of Medicine. For the past nine years, he has been involved in university efforts to address public health needs in the Philadelphia area through programs involving medical students. PMID- 7478669 TI - Office managers: nobody knows the trouble they see. AB - Computers, code revisions, and patients' changing expectations have added responsibility and a new level of complexity to the job of medical office manager. How are you and your office manager coping? Some experienced managers relay their views of this rapidly changing field. PMID- 7478670 TI - Hold harmless, due process resolutions supported by AMA House of Delegates. PMID- 7478671 TI - Preventing a fall. PMID- 7478672 TI - Dealing with--and learning from--the uncomfortable interview. PMID- 7478673 TI - Supporting nurses: safety and quality initiatives of the American Nurses Association. PMID- 7478674 TI - A look at case management certification. PMID- 7478675 TI - Female alcoholics more adversely affected by alcohol. PMID- 7478676 TI - Negotiating the raise you deserve. PMID- 7478677 TI - Insights into the development of opioid tolerance. PMID- 7478678 TI - Experimental mononeuropathy reduces the antinociceptive effects of morphine: implications for common intracellular mechanisms involved in morphine tolerance and neuropathic pain. AB - Recent evidence suggests that hyperalgesia and morphine tolerance, two seemingly unrelated phenomena, have in common certain neural substrates such as activation of the N-methyl-D-aspartate (NMDA) receptor and the subsequent intracellular activation of protein kinase C and nitric oxide. Should common cellular elements be involved in hyperalgesia and morphine tolerance, these cellular and intracellular commonalities might be expected to result in interactions between these two phenomena. Indeed, our previous studies have shown that thermal hyperalgesia develops when animals are made tolerant to the antinociceptive effects of morphine. In this study, we examined the hypothesis that reduction of morphine antinociception occurs following unilateral ligation of the rats's sciatic nerve, a procedure which produces symptoms of a neuropathic pain syndrome including thermal hyperalgesia. When tested using the paw-withdrawal test on day 8 (D8) after either nerve ligation or sham operation, a single intrathecal treatment with 10 micrograms morphine sulfate (30 min after administration) produced significant antinociception in sham-operated rats but not in nerve injured ones. These results also were obtained when thermal hyperalgesia was reversed in nerve-injured rats by the non-competitive NMDA receptor antagonist MK 801. Consistently, 8 days after sciatic nerve ligation but not after a sham operation, an approximately 6-fold rightward shift occurred in the morphine antinociceptive dose-response curve. This rightward shift of the morphine antinociceptive dose-response curve did not occur at 24 h after either nerve ligation or sham operation. In addition, once daily treatment with 10 nmol MK-801 from D2 to D7 after nerve ligation prevented both the development of thermal hyperalgesia and the rightward shift of the morphine antinociceptive dose response curve on D8. The results indicate that the antinociceptive effects of morphine are reduced in nerve-injured rats in the absence of daily exposure to morphine and that the NMDA receptor activation may have a critical role in mechanisms of this phenomenon. These data provide further evidence indicating that interactions do occur between neural mechanisms underlying thermal hyperalgesia and morphine tolerance. PMID- 7478679 TI - The development of morphine tolerance and dependence is associated with translocation of protein kinase C. AB - The development of tolerance to the analgesic effects of morphine as well as morphine dependence were greatly reduced by co-administration with morphine of GM1 ganglioside, a substance reported to block the translocation of protein kinase C (PKC) from cytosol to membrane of neurons. Rats made tolerant to intrathecal administration of morphine showed increased membrane-bound PKC in the superficial layers (laminae I and II) of the spinal cord dorsal horn but not in deeper layers. This increase was prevented by co-administration with morphine of GM1 ganglioside. These results indicate that the translocation and activation of PKC may be a critical step in the development of opiate tolerance and dependence. Modulation of PKC translocation and activation may prove useful for the management of pain and opiate addiction. PMID- 7478680 TI - Chronopharmacokinetic variability in plasma morphine concentrations following oral doses of morphine solution. AB - Twenty-six patients with severe pain associated with cancer were entered into a study where they were required to take morphine mixture for 7 days. Prior to this, their morphine dose had been optimised to provide the most favourable balance between pain relief and side effects. After 6 days of taking their optimised morphine dose at 4-hourly intervals, the patients were admitted to the Pain Management Unit such that the doses from 18:00 h on day 6 were taken under direct nursing supervision. Frequent blood samples were collected after the 10:00 h (dose 1), 14:00 h (dose 2) and 18:00 h (dose 3) on day 7. There was a significant difference between the 3 doses with respect to Cmax values for morphine with dose 3 > dose 1 > dose 2. Further, there was considerable variability in the percentage change of either dose 1 or dose 3 Cmax values relative to dose 2. The Cmax values of the active metabolite morphine-6 glucuronide (M6G), measured in 6 of the patients, for the 3 dosing intervals followed a similar trend to the parent drug, but only doses 1 and 2 differed significantly. Similar but less pronounced changes were observed in the area under-curve parameter calculated for both morphine and M6G during the 3 dosing intervals. There were no significant differences in the Cmin or Tmax parameters for either morphine or M6G between the 3 dosing intervals. These results suggest intra-individual variation in the absorption of morphine or changes in the volume of distribution during the day.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7478681 TI - Noxious stimuli induce an N-methyl-D-aspartate receptor-dependent hypersensitivity of the flexion withdrawal reflex to touch: implications for the treatment of mechanical allodynia. AB - Primary C-fiber afferents can induce a state of increased excitability in spinal cord neurons amplifying their responsiveness to noxious and innocuous stimuli- the phenomenon of central sensitization. We have examined whether the hypersensitivity to low-intensity stimuli (mechanical allodynia) evoked by C afferent conditioning inputs is NMDA receptor dependent. The enhancement by C afferent conditioning stimuli of the normally low or absent cutaneous touch evoked responses of posterior biceps femoris/semitendinosus flexor motoneurons in the decerebrate-spinal rat has been used as a model of touch-evoked allodynia. Three different noxious conditioning stimuli (electrical stimulation of the sural nerve at C-fiber strength, cutaneous application of mustard oil and the intramuscular (i.m.) injection of bradykinin) enhanced touch-evoked responses and decreased the threshold of the flexor reflex to mechanical stimulation. Intrathecal (i.t.) injections of NMDA (20 pmol) produced a similar effect. A non competitive NMDA antagonist MK 801, administered prior to the conditioning input at doses that did not modify the baseline reflex, significantly attenuated both the enhancement of touch-evoked responses and the threshold decrease following the noxious conditioning stimuli. Treatment with MK 801 some time after the conditioning input when the mechanical hypersensitivity is fully established, also reduced the increased touch-evoked responses. The reduction in threshold and the increase in touch responsiveness induced by cutaneous and muscle noxious C fiber conditioning stimuli in the rat spinal cord are, therefore, both prevented and reversed by NMDA receptor antagonism. NMDA antagonists may be potentially useful, therefore, in treating postinjury pain hypersensitivity. PMID- 7478683 TI - Dextromethorphan suppresses both formalin-induced nociceptive behavior and the formalin-induced increase in spinal cord c-fos mRNA. AB - The injection of dilute formalin results in a stereotyped nociceptive behavioral response. Administration of dextromethorphan (s.c.) but not saline, 30 min prior to intraplantar formalin injection prevents this nociceptive response in a dose dependent manner. In addition, intraplantar formalin reliably induces c-fos mRNA in the ipsilateral spinal dorsal horn as assessed with quantitative solution hybridization at 30 min postinjection. No change in c-fos mRNA was detected in the contralateral spinal dorsal horn, nucleus raphe magnus, periaqueductal grey, medial thalamus, or sensorimotor cortex. Pretreatment with dextromethorphan at 60 mg/kg s.c., 30 min prior to formalin resulted in a suppression of c-fos induction, so that c-fos mRNA levels in the ipsilateral spinal dorsal horn of animals receiving dextromethorphan prior to formalin did not differ from controls. These data indicate that dextromethorphan suppresses formalin nociceptive behavior and one of the biochemical consequences of formalin nociception, i.e., induction of c-fos mRNA. PMID- 7478682 TI - Epidural clonidine analgesia for intractable cancer pain. The Epidural Clonidine Study Group. AB - Although the vast majority of patients with cancer pain receive effective analgesia from standard therapy, a few patients, particularly those with neuropathic pain, continue to experience severe pain despite large doses of systemic or intraspinal opioids. Animal studies suggest intraspinal alpha 2 adrenergic agonists may be effective in such cases. Eighty-five patients with severe cancer pain despite large doses of opioids or with therapy-limiting side effects from opioids were randomized to receive, in a double-blind manner, 30 micrograms/h epidural clonidine or placebo for 14 days, together with rescue epidural morphine. Pain was assessed by visual analog score (VAS), McGill Pain Questionnaire, and daily epidural morphine use. Success was defined as a decrease in either morphine use of VAS pain, with the alternative variable either decreasing or remaining constant. Blood pressure, heart rate, and degree of nausea and sedation were monitored. Successful analgesia was more common with epidural clonidine (45%) than with placebo (21%). This was particularly prominent in those with neuropathic pain (56% vs. 5%). Pain scores were lower at the end of the treatment period in patients with neuropathic pain treated with clonidine rather than placebo, whereas morphine use was unaffected. Clonidine, but not placebo, decreased blood pressure and heart rate. Hypotension was considered a serious complication in 2 patients receiving clonidine and in 1 patient receiving placebo. This study confirms the findings from previous animal studies which showed the effective, potent analgesic properties of intraspinal alpha 2 adrenergic agonists and suggests that epidural clonidine may provide effective relief for intractable cancer pain, particular of the neuropathic type. PMID- 7478684 TI - Infusion of substance P or neurokinin A by microdialysis alters responses of primate spinothalamic tract neurons to cutaneous stimuli and to iontophoretically released excitatory amino acids. AB - The responses of 25 spinothalamic tract (STT) neurons to mechanical and thermal stimulation of the skin, as well as to a battery of iontophoretically applied excitatory amino acids (EAAs), were tested before and then during microdialysis of substance P (SP) or neurokinin A (NKA) into the dorsal horn of anesthetized monkeys. Neither peptide had significant effects on the background activity or the responses to mechanical or thermal stimulation of the skin. However, each peptide produced significant increases in the responses to iontophoretic application of one or more EAAs. In addition, following combined application of the EAAs and either SP or NKA, the responses of the cells to mechanical stimulation of the skin increased. Combined application of SP and NKA failed to produce an increase in responses to either the EAAs or to cutaneous stimuli that was greater than that produced by either peptide alone. It is concluded that SP and NKA produce an increase in the responses of STT cells to iontophoretic applications of EAAs and the combined effects of these compounds produce sustained increases in responses to mechanical stimulation of the skin. These changes mimic those observed when STT cells are sensitized by peripheral noxious stimuli, suggesting that the mechanism of induction and expression of sensitization involves the facilitation of dorsal horn neuron responses to EAAs by tachykinins. PMID- 7478685 TI - The Pain Beliefs and Perceptions Inventory: a British replication. AB - The factorial structure, reliability, and validity of the Pain Beliefs and Perceptions Inventory (PBPI) was investigated in a sample of 84 pain patients drawn from a pain clinic in the United Kingdom. The recovered factorial structure replicated that of a previous study. The 4 derived subscales, Mysteriousness, Self-Blame, Pain Constancy and Pain Permanence, had excellent reliability (internal consistency); Chronbach's alpha was greater than 0.80. However, the construct validity of the PBPI, assessed by examining the association between the subscales and the Coping Strategies Questionnaire (CSQ), was not supported. The results are discussed with particular reference to the interpretative issues concerning 2 of the subscales and their relation to the concept of the 'time line' component of illness representation. PMID- 7478686 TI - Gender differences in pain ratings and pupil reactions to painful pressure stimuli. AB - In order to investigate gender differences in pain perception, the present study employed both a psychophysical and a psychophysiological measure. In experiment 1, 20 subjects rated the painfulness of 4 different levels of tonic pressure applied to their fingers using a verbally anchored categorization procedure. In general agreement with studies of pain threshold and tolerance, female subjects reported greater pain at high levels of stimulation, with no gender difference being evident at low pressure levels. In experiment 2, 16 different subjects were exposed to the same painful pressure stimuli while measuring their pupil reactions using infrared video pupillometry. The pupil dilations seen during the last 10 sec of the 20-sec pressure application turned out to be a highly significant indicator of pain intensity. When female and male subjects were compared on this measure, a similar divergent pattern as in the psychophysical data emerged, with female subjects showing greater pupil dilations at high pressure levels only. The fact that gender differences in pain perception can be demonstrated using an autonomic indicator of pain that is beyond voluntary control suggests that these differences reflect low-level sensory and/or affective components of pain rather than attitudinal or response-bias factors. PMID- 7478687 TI - A multicenter randomized controlled trial of injections of sterile water and saline for chronic myofascial pain syndromes. AB - A randomised controlled trial was used to test the hypothesis that cutaneous injections of sterile water (SWI) have no benefit over saline (PSI) as a method of pain reduction among patients with myofascial pain syndromes. Six general practitioners located at 6 different clinics of general practice treated 117 patients (91 female, 26 male, aged > or = 25 years) with myofascial pain syndrome for at least 3 months in one or both of the upper quadrants of the body. Patients were randomised to receive either SWI or PSI which was administered sub- and intracutaneously on 1 occasion. The patients received a mean number of 10 injections of 0.5 ml of either substance. The main outcome measure was pain intensity which was measured with visual analogue scales before intervention, 10 min after intervention and 14 days after intervention. We found no statistically or clinically significant difference in pain level reduction between the 2 groups. However, patients who received SWI reported a much more painful treatment experience than those who received PSI. Our study shows that injections of sterile water are substantially more painful but demonstrate no better clinical outcome than similar injections of saline as a method to treat patients with chronic myofascial pain syndrome. PMID- 7478688 TI - A randomized controlled trial of citalopram in the treatment of fibromyalgia. AB - Amitriptyline and cyclobenzaprine have shown some efficacy in treatment of the generalised pain syndrome, fibromyalgia. The aim of this study was to examine the efficacy of antidepressant dosages of the serotonin re-uptake inhibitor citalopram in fibromyalgia. In a double-blind, placebo-controlled study 22 patients with fibromyalgia were randomized to treatment with citalopram for 4 weeks at a dosage of 20 mg a day while 21 received placebo. After 4 weeks the dosage was increased to 40 mg for a further 4 weeks if the subjects did not report a marked improvement. After the end of treatment (8 weeks) no changes were observed in self-assessment of symptoms, physician's global assessment, tender points, Beck depression score or voluntary muscle strength and no differences were observed between the groups. Citalopram showed no demonstrable effect on this group of pain patients. The strength of the study was sufficient to exclude an effect of citalopram of more than 1 steps of 10 on the categoric scales for pain, fatigue and general condition (95% confidence limit), which indicates that the sample size was sufficiently large. PMID- 7478689 TI - Musculoskeletal pains and depression in a middle-aged Finnish population. AB - The relationships between depression, measured as high rates of depressive symptoms (Zung Self-Rating Depression Scale, ZSDS), and musculoskeletal pains (Kuorinka et al. 1987) were described in a 55-year-old Finnish population consisting of all the 1008 persons born in 1935 and living in the city of Oulu on 1 October 1990. Three hundred forty-five men (76%) and 435 women (79%) participated in the examinations. Of the men 6.8% and of the women 12.1% scored 45 raw sum points or more on the ZSDS. In several anatomical regions, pains were more common among the depressed than the non-depressed population, and many of the depressed persons suffered from multiple pains. One of the most common regions of pain was the neck; during the past 12 months, 56.5% of the depressed men and 65.4% of the depressed women had suffered from frequent pains in the neck. The corresponding prevalences among the non-depressed men and women were 35.2% and 45.5%, respectively. In the non-depressed population, musculoskeletal pains were more common among women than men, whereas no great gender differences existed in the depressed population. The possible confounding variables were standardized in the logistic regression analysis, and the results showed an independent association between pains in the small joints and depression among men, on one hand, and pains in the neck and shoulder and depression among women, on the other hand. PMID- 7478690 TI - Artificial ureteral calculosis in rats: behavioural characterization of visceral pain episodes and their relationship with referred lumbar muscle hyperalgesia. AB - In a rat model of artificial ureteral calculosis, the aim of the study was to characterize the behavioural manifestations of direct visceral pain and to evaluate the relationship between number, duration and complexity of the visceral episodes and the extent of referred lumbar muscular hyperalgesia. As evidenced by non-stop video-tape recordings over 4-14 days, almost 98% of stone-implanted rats showed episodes similar to the writhing behaviour characteristic of noxious visceral stimulation in animals. From one rat to another, these episodes varied from very few (1-3) to a very high number (+/- 60), lasted a few minutes to over 45 min and were of variable complexity, as evaluated via an arbitrary scale on the basis of the combination of movements. Their number and duration decreased significantly, in a linear fashion, as time passed after the operation, so that they were mostly concentrated during the first 3 days. Number, duration and complexity of episodes were reduced by chronic treatment with morphine in a dose dependent fashion. Stone-implanted rats displaying visceral episodes also showed hyperalgesia of the ipsilateral oblique musculature, as evidenced by a decrease in the vocalization threshold to electrical muscle stimulation, which was maximum on the first 3-4 days after implantation but lasted up to 10 days. The visceral episodes and the muscle hyperalgesia showed a strict relationship of interdependence: a significant, direct linear correlation was found between number and duration of episodes and tendency to also develop a contralateral muscle hyperalgesia. By applying the results of the study to the interpretation of human pathology, referred lumbar muscle hyperalgesia from ureteral calculosis would appear to be a strict function of the colic pain experienced. PMID- 7478691 TI - Differential response to pain by very premature neonates. AB - The ability of very low birth weight (VLBW) premature infants to respond differentially to real versus a sham heelstick conditions was examined in this crossover study. Using a multidimensional assessment of responses of premature infants (n = 48) between 26 and 31 weeks gestational age (GA) at the time of the study, it was found that they respond differentially to real versus sham heelstick both behaviourally and physiologically. The multivariate effect of condition (real/sham) was significant with maximum heart rate and upper facial action (lower facial action was not scored) contributing significantly to the main effect. GA had a main multivariate effect, with the younger infants responding less robustly. The variability outcome measures of heart rate standard deviation and range of transfontanelle intracranial pressure contributed significantly to the main effect of GA, but not to the effect of condition. Young VLBW premature infants are capable of a multidimensional differential response to pain. GA is an important factor to consider when assessing pain in premature infants. PMID- 7478692 TI - Modulation of pressure pain thresholds during and following isometric contraction. AB - This study aimed at evaluating the influence of submaximal isometric contraction on pressure pain thresholds (PPTs) in 14 healthy volunteers before and after skin hypoesthesia. PPTs were determined with pressure algometry over m. quadriceps femoris before, during, and following an isometric contraction. Maximum voluntary contraction (MVC) was assessed using a computerized dynamometer. A contraction of 21% MVC was held until exhaustion (max: 5 min) and PPTs were assessed every 30 sec. A local anesthetic cream and a control cream were applied following a double blind design and PPTs were reassessed. PPTs increased significantly at the start of contraction and continued to increase until the middle of the contraction period, then remaining at this level. After contraction PPTs decreased significantly but for 5 min remained slightly above precontraction levels. Anesthetic cream raised PPT at rest but not during and following contraction. The relative increase in PPTs during and immediately following isometric contraction was lower with anesthetic cream. Isometric contraction of m. quadriceps femoris increase PPTs during and following contraction. The results suggest that input from cutaneous and deeper tissues interacts with nociceptive activity set up by the pressure stimulus. Determining the degree of sensory modulation in muscle and skin in different chronic pain syndromes could become a functional method of patient assessment important for differential diagnosis, treatment evaluation, and follow-up. PMID- 7478693 TI - Comments on Barnsley et al. PMID- 7478694 TI - Evaluation of life expectancy in selection of patients undergoing percutaneous cervical cordotomy or subarachnoid phenol block for pain control of costopleural syndrome. PMID- 7478695 TI - Asthma of the back. PMID- 7478696 TI - Expansion and comments on non-steroidal anti-inflammatory drugs and spinal nociception processing (Pain, 59 (1994) 9-43) PMID- 7478697 TI - Pediatric oncology procedures: to sleep or perchance to dream? PMID- 7478698 TI - The factorial structure and stability of the McGill Pain Questionnaire in patients experiencing oral mucositis following bone marrow transplantation. AB - The McGill Pain Questionnaire (MPQ) (Melzack 1975) is an important assessment tool for multidimensional pain measurement in both clinical practice and research. Despite widespread acceptance, empirical analyses have not consistently verified the 3 a-priori factors that guided the subclass construction of the Pain Rating Index (PRI) of the MPQ. This study compared the a-priori model with 2 qualitatively different factor models in 191 patients with oral mucositis pain at 3 days and 10 days following bone marrow transplantation. A semantic model defined by Sensory Action, Sensory Evaluation, and Affective Evaluation factors of subclass descriptor content fit better than the a-priori model and a model positing a single general pain factor. The 3 semantic PRI factors were highly intercorrelated, with the sensory factors correlating more highly with an independent visual analogue (VAS) pain scale. Standardized factor regression coefficients between the two occasions of measurement ranged between 0.4 and 0.5. Mean factor change was greatest for Sensory Evaluation and lowest for Affective Evaluation. All analyses were conducted with the LISREL 7 structural equation modeling program. Although the factor analyses indicated an unambiguous ranking of PRI models according to statistical criteria, these theoretical results generalize poorly to simple scores formed by direct addition of the PRI subclasses. Summary scores can only approximate the unobserved factors and cannot retain the fine discriminations revealed by the theoretical factors. Psychometric considerations suggest that a single PRI total score will yield better practical measurement than any scoring rules based on multiple factors. PMID- 7478699 TI - General and segmental reduced pain thresholds in juvenile chronic arthritis. AB - The objective of this work was to investigate the pattern of pain threshold (PT) alterations in paraspinal soft tissues as related to inflamed joints in patients with juvenile chronic arthritis (JCA). A pressure algometer was used in JCA patients with inflamed knee(s) (n = 16), with inflamed ankle(s) (n = 17), and in healthy controls (n = 69), all aged 6-17 years. Local pressure was applied to the joint capsules of the knees and ankles and the soft paraspinal tissues, and PTs were recorded. JCA patients showed not only a significantly lower PT at the inflamed knee and ankle joints, but also in the non-inflamed paraspinal areas. All paraspinal PTs showed a significant negative correlation with the inflamed knee or ankle joint. The correlation rank order showed that the paraspinal L1 area had the highest negative correlation with the inflamed knee joint and the paraspinal L3 area with the inflamed ankle joint. JCA coincides with generally diminished pain thresholds in the paraspinal region. Nociception from inflamed joints may have established changes in the peripheral as well as central nociceptive processing system in JCA. PMID- 7478700 TI - Lack of analgesic effect of 50 and 100 mg oral tramadol after orthopaedic surgery: a randomized, double-blind, placebo and standard active drug comparison. AB - Tramadol hydrochloride is a synthetic mu-opioid agonist with additional monoaminergic activity. Tramadol's analgesic effect has been equated with that of pethidine, with a more favourable side-effect profile. Tramadol has been the most selling prescription analgesic in Germany for several years, and it is now available in many other European countries, but still there is a lack of adequately controlled clinical studies of its analgesic properties. The purpose of this study was to compare the analgesic efficacy of 50 and 100 mg oral tramadol with our standard analgesic for postoperative pain treatment, 1000 mg paracetamol + 60 mg codeine, and placebo. A single-dose, parallel group, double blind design was used. One hundred forty-four patients were enrolled the day after total hip replacement if they had a pain intensity of 60 mm or more on a 0 100 mm visual analogue scale. Treatments were compared on the basis of pain intensity and derived variables (pain intensity difference, and summed pain intensity differences), the need of rescue medication, and a global evaluation. Serum concentrations confirmed rapid and good absorption comparable with that reported in healthy volunteers. The active drug control, paracetamol+codeine, was significantly superior to placebo for all efficacy variable (P = 0.0002-0.004), confirming good assay sensitivity. Paracetamol+codeine was also significantly superior to both 50 mg tramadol (P = 0.002-0.03) and 100 mg tramadol (P = 0.002 0.02). There was no difference between placebo and 50 and 100 mg tramadol for any of the efficacy variables.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7478701 TI - Ketamine as an adjunct to morphine in the treatment of pain. AB - A double-blind multidose trial of the addition of ketamine (0-40 mg, i.m., 8 times per day) to intramuscular morphine therapy was undertaken in a 61-year-old man with chronic back pain related to osteoporosis who had received inadequate pain relief from anterior interbody fusion, dorsal column stimulation and morphine alone. The patient reported only mild side effects. Nausea, tiredness and well-being were not significantly influenced by the ketamine dose level. Visual analogue pain scores prior to each dose were not associated with the ketamine dose level, but pain scores 30 min after doses were significantly reduced in a dose-related manner. In addition, the amount of morphine used by the patient was significantly reduced as the ketamine dose increased. This patient experienced substantial benefit from the addition of ketamine to intramuscular morphine therapy. PMID- 7478702 TI - What is a nociceptor-specific (class 3) cell? PMID- 7478703 TI - Comment on D.G. Hawley and F. Wolfe. PMID- 7478704 TI - Comment on C.C. Gunn--re: fibromyalgia. PMID- 7478705 TI - Responses of on and off cells in the rostral ventral medulla to stimulation of vagal afferents and changes in mean arterial blood pressure in intact and cardiopulmonary deafferented rats. AB - The relationships between mean arterial blood pressure (MAP) and the activity of putative pain modulatory neurons of the rostroventral medulla (ON and OFF cells) were determined in intact and cardiopulmonary deafferented rats. A total of 173 neurons were recorded from 97 rats as follows: 32 ON cells and 25 OFF cells from 39 intact rats; 32 ON cells and 20 OFF cells from 24 rats with bilateral sino aortic deafferentation (SAD); 12 ON cells and 20 OFF cells from 19 rats with bilateral cervical vagotomy (CVAG); and 20 ON cells and 12 OFF cells from 15 rats with both SAD and CVAG. ON and OFF cells showed spontaneous fluctuations in activity such that ON cell activity was negatively correlated with MAP whereas OFF cell activity was positively correlated with MAP under conditions of no applied stimuli. These correlations were present in both intact and cardiopulmonary deafferented rats. Further, experimentally induced increases in MAP decreased ON cell activity and increased OFF cell activity in intact rats, but not in rats with SAD, CVAG, or the combination of SAD and CVAG. Experimentally induced decreases in MAP decreased OFF cell activity in intact rats and rats with CVAG, but not in rats with SAD or the combination of SAD and CVAG. These findings indicate that ON and OFF cells are modulated by baroreceptor activity, but baroreceptor input is not necessary for the spontaneous fluctuations in ON and OFF cell activity. Electrical stimulation of vagal afferents (VAS) inhibited 60% of the OFF cells studied, excited 4%, and produced biphasic effects consisting of excitation at low intensities and inhibition at greater intensities in 28% of all OFF cells. In general, VAS excited the majority of the ON cells studied, although there were significant differences between effects in intact and cardiopulmonary deafferented rats. Greater intensities of VAS that inhibited OFF cells and excited ON cells also inhibited the tail flick. Thus, inhibition of OFF cells and excitation of ON cells was correlated with antinociception. The effects of intravenous (i.v.) administration of 1.0 mg/kg morphine on neuronal activity did not differ between intact and cardiopulmonary deafferented rats.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 7478706 TI - A comparative study of cognitive behavior therapy versus general anesthesia for painful medical procedures in children. AB - A treatment outcome study was conducted to compare the efficacy of cognitive behavior therapy (CBT) versus general anesthesia in alleviating the distress of 18 pediatric cancer patients (ages: 3-12 years) undergoing bone marrow aspirations (BMAs). CBT and short-acting mask anesthesia were delivered within a repeated-measures counterbalance design. Results indicated that children exhibited more behavioral distress in the CBT condition for the 1st minute lying down on the treatment table. However, parents rated significantly more behavioral adjustment symptoms 24 h following the BMA when their children had received anesthesia. No differences were found in childrens' and parents' preference for CBT versus anesthesia. PMID- 7478707 TI - Scaling the affective domain of pain: a study of the dimensionality of verbal descriptors. AB - This study evaluated the multidimensional structure of affective verbal descriptors and investigated individual differences in the scaling of the descriptors. Patients with chronic low back pain, chronic headache and rheumatoid arthritis (25 per group) and 25 control subjects, matched for age and sex, made similarity judgements of a set of 12 verbal descriptors, e.g., awful, miserable. They also completed the Coping Strategies Questionnaire, the Hospital Anxiety and Depression Scale, McGill Pain Questionnaire Short Form, and a measure of verbal intelligence. The similarity data were subjected to individual differences multidimensional scaling (ALSCAL) and the resulting subject weights were related to group membership and the questionnaire data. The multidimensional nature of affective descriptors was confirmed. A 4-dimensional solution was identified: the first 3 dimensions were comparable with previous findings and labeled Tolerability, Focus of Attention and Minor Emotional Reaction. The 4th dimension was enigmatic and attributable to the back pain group. There were significant differences between the groups in their weighting of the dimensions and in their self-reported coping strategies. The results are discussed with reference to an earlier study and the degree of consistency across the studies is noted. Implications of the results for the conceptualisation and measurement of the affective domain of pain report are outlined. PMID- 7478708 TI - Morphine-3-glucuronide: evidence to support its putative role in the development of tolerance to the antinociceptive effects of morphine in the rat. AB - Antinociceptive tolerance to morphine (MOR) was induced in groups of Sprague Dawley rats receiving continuous intravenous infusions of morphine sulphate administered by 3 different MOR dosing regimes. At appropriate intervals throughout each infusion period, antinociceptive testing was performed using the tail-flick latency test and blood samples were collected. Groups of saline (SAL) infused control rats also underwent antinociceptive testing and blood sample collection. Complete antinociceptive tolerance developed during each MOR infusion period and was characterized by a marked decline in the degree of antinociception from values greater than 90% of the maximum possible effect (%MPE) to pre-dosing baseline values. By contrast, %MPE values in SAL-infused control animals and in sham-operated rats were not significantly different from pre-dosing values throughout the infusion period, indicating that the experimental procedures themselves did not contribute to the development of antinociceptive tolerance to MOR. In addition, the rate of MOR tolerance development was inversely proportional to the MOR infusion rate. A very significant inverse relationship was observed between the mean degree of antinociception (%MPE) and the mean plasma molar concentration ratio, [morphine-3-glucuronide]/[MOR], for each of the 3 MOR dosing regimes and for the cumulated data. This relationship showed that near-maximum antinociception was attainable at ratio values less than approximately 0.50, whilst at ratio values above approximately 1.5, little or no antinociception was observed. Although %MPE was highly inversely correlated with the mean plasma morphine-3-glucuronide (M3G) concentrations for rats receiving regimes A and B, this was not the case for rats receiving regime C where antinociceptive tolerance was partially reversed by an increase in the morphine infusion rate part-way through the infusion period. In addition, a poor relationship was observed between %MPE and the mean plasma MOR concentration, possibly due to the confounding presence of M3G in all samples. Thus, we may conclude from this study in Sprague-Dawley rats that irrespective of the rate of antinociceptive tolerance development, the level of antinociception achievable appears to be highly inversely correlated with the mean [M3G]/[MOR] plasma molar concentration ratio and poorly correlated with the plasma MOR concentration, consistent with the notion that it is perhaps the balance between the excitatory effects of M3G and the inhibitory effects of MOR at the functional level which is the important determinant. Further research is required in carefully conducted studies in cancer patients to evaluate the possible contribution of the MOR metabolites, M3G and morphine-6-glucuronide (MbG), to increasing dosing requirements of MOR.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 7478709 TI - Parabrachial area and nucleus raphe magnus-induced modulation of electrically evoked trigeminal subnucleus caudalis neuronal responses to cutaneous or deep A fiber and C-fiber inputs in rats. AB - The aim of this study was to determine whether trigeminal subnucleus caudalis (Vc) neurons show differential inhibitory effects produced by parabrachial area (PBA) or nucleus raphe magnus (NRM) conditioning stimulation (CS) on their A fiber versus C-fiber inputs electrically evoked from cutaneous and deep tissues. A total of 55 Vc neurons in urethane/chloralose-anesthetized rats were functionally classified as low-threshold mechanoreceptive (LTM; n = 12), nociceptive (wide-dynamic-range, WDR, and nociceptive-specific, NS; n = 11), deep (D; n = 6), or nociceptive convergent neurons receiving skin and deep inputs (S+D; n = 26). Most neuronal responses were tested for both PBA and NRM CS (20-40 microA) delivered 50 msec prior to test stimulation (1.1-1.5 x threshold) of their mechanoreceptive field (RF) or the hypoglossal nerve (XII). Inhibition of responses to both cutaneous and deep A-fiber and C-fiber inputs occurred in over two-thirds of the Vc neurons tested in each neuronal class, and facilitation of responses was only occasionally found. In general, both PBA (ipsi- and contralateral) and NRM CS produced profound inhibition of electrically evoked responses in all neuron types, e.g., responses reduced to mean values of 50-53% of control in LTM neurons, 35-46% in WDR, NS, and S+D neurons, and 61-63% in D neurons. No significant difference in the magnitude of their inhibitory effects was found between PBA and NRM, and between Vc neuronal responses to A- and C fiber inputs irrespective of their origin from skin or deep tissues.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7478710 TI - The use of topical 4% lidocaine in spheno-palatine ganglion blocks for the treatment of chronic muscle pain syndromes: a randomized, controlled trial. AB - To assess the efficacy of 4% topical lidocaine in spheno-palatine blocks, a randomized controlled trial was carried out on patients with chronic muscle pain syndromes. Sixty-one patients (42 with fibromyalgia (FM) and 19 with myofascial pain syndrome (MPS)) completed the trial. Outcome measures included pain intensity, a daily pain diary, headache frequency, sensitivity to pressure using a dolorimeter, anxiety, depression, and sleep quality. Patients were randomized to receive either 4% lidocaine or sterile water (placebo) 6 times over a 3-week period. Both subjects and investigators were blind to treatment allocation. The results showed that 4% lidocaine had no superiority over placebo in any of the outcome measures. Twenty-one subjects (35%) showed a decrease in pain which was greater than 30% of their baseline value. Of these 21 subjects, 10 received lidocaine and 11 received placebo. These data suggest that, in this population, 4% lidocaine is no better than placebo in the treatment of chronic muscle pain. PMID- 7478711 TI - The role of coping in adjustment to phantom limb pain. AB - Research has indicated that the way individuals cope with pain may influence pain, and physical and psychological adjustment. The present study assessed the relationship between coping strategy use and adjustment in amputees with phantom limb pain (PLP). Coping strategies were measured using the Coping Strategies Questionnaire (CSQ) and their relationship with adjustment was examined using both composite scores and individual strategy scores. The results indicated that the relationship between coping and adjustment was primarily explained by the use of strategies within the 'Helplessness' factor of the CSQ. A clearer picture of coping strategy use was gained from examining individual strategies rather than the composite measures. Catastrophizing explained the largest proportion of variance in pain report (26%), while increasing behavioral activity and hoping or praying strategies explained a smaller proportion of the variance (3% and 1%, respectively). Catastrophizing also explained a large proportion of the variance in physical and psychosocial dysfunction (11% and 22%), while hoping or praying strategies accounted for only a small proportion of the variance in physical dysfunction (3%), and re-interpreting pain sensations accounted for a small proportion of the variance in psychosocial dysfunction (3%). The findings in this study have important clinical implications in that coping strategy use was associated with increased, rather than decreased, levels of pain and disability. However, since the reported use of coping strategies in the present study was low, further research, perhaps utilizing other measures of coping, is required to clarify these findings. PMID- 7478712 TI - Effects of oxygen radicals on nociceptive afferents in the rat skin in vitro. AB - On the premises of clinical studies, a possible contribution of oxygen radicals has been considered to the development of inflammatory pain and hyperalgesia. In a rat skin-saphenous nerve preparation using standard teased-fiber techniques (n = 57) hydrogen peroxide (1 mM, 10 mM and 50 mM) was applied in aqueous solution to cutaneous nerve endings of unmyelinated nociceptive afferents. Superoxide anion and hydroxyl radical were secondarily generated as reaction products from pyrogallol (1 and 10 mM) and from Fe-EDTA (1 mM) in hydrogen peroxide, respectively. None of these substances, except exceptionally, induced ongoing activity nor nociceptor sensitization to heat and mechanical stimuli. If occasionally there was a weak excitatory effect, the fibers were left with a profound desensitization to adequate stimulation. The addition of hydrogen peroxide did not enhance sustained responses to solutions of high proton concentration (pH 6.1). Responses to combined inflammatory mediators (bradykinin, serotonin, histamine and prostaglandin E2, 10 microM) were increased, on average, when hydrogen peroxide was added but this effect did just not reach significance. These findings suggest that oxygen radicals do not play a major and specific role in nociceptor sensitization. PMID- 7478714 TI - Treatment of Cushing's disease by interstitial pituitary irradiation: short- and long-term follow-up. AB - Trans-sphenoidal intrasellar implantation of radioactive rods was employed to treat Cushing's disease in our Institution between 1958 and 1981. The patients were followed at regular intervals after the procedure. The aim of this work is to assess retrospectively the results, comparing the short- (1 year) and long term (average 21.8 years) effects of this treatment. Seventy-six patients received pituitary implantation of 90Y- and one of 198Au-labelled rods, delivering a dose of 100-150,000 rad. Complete remission was obtained in a few weeks to months in 57/76 patients (5 of whom required a second implantation); 2 patients died of meningoencephalitis and 3 of cardiovascular complications associated with hypercortisolism 1 to 2 months after surgery. In 12 patients bilateral adrenalectomy or external pituitary irradiation were required to achieve remission; one of them developed Nelson's syndrome 15 years after implantation. Two were lost at follow-up. Long-term follow-up was possible in 41 patients of the initial series. Of these, 40 were cured of the disease, with hypoadrenalism developing in 25, while recurrence was observed only in the patient treated with 198Au. The incidence of hypothyroidism was 50%, and that of hypogonadism 54%. Permanent diabetes insipidus developed in 1 subject. GH deficiency resulting in retarded growth was found in the youngest patient, who had been operated at the age of 14. In conclusion, interstitial irradiation of pituitary adenomas was a safe and effective procedure for the treatment of Cushing's disease. PMID- 7478713 TI - The influence of lockout intervals and drug selection on patient-controlled analgesia following gynecological surgery. AB - This study systematically compared 2 opioids, morphine (MOR) and fentanyl (FEN), and 2 lockout intervals, long (L) and short (S) in patients utilizing patient controlled analgesia (PCA). Seventy-eight women undergoing gynecological surgery were randomly assigned to 1 of 4 groups: MOR-S (7 min), MOR-L (11 min), FEN-S (5 min), FEN-L (8 min). PCA measures obtained during the first 24 h after surgery included: number of demands/h, number of completed deliveries/h, dose/h, and demand/delivery ratio. Visual analog scales of pain and anxiety were also obtained. Results indicated that pain relief was equivalent with minimal side effects for both opioids. The selection of opioid, however, influenced the pattern of PCA use, with an improved demand/delivery ratio initially for FEN. The lockout intervals chosen for this study did not influence pain or anxiety levels. PMID- 7478715 TI - Preoperative mapping of the saphenous vein: prognostic value on early postoperative results. AB - A consecutive series of 92 patients had their greater saphenous vein assessed with duplex ultrasound scanning prior to planned infrainguinal bypass procedures. A naturally occurring optimal vein diameter was discovered. It was significantly correlated with higher postoperative ankle-brachial pressure index (ABI) and lesser early postoperative thrombosis. A significant linear regression was found between the pre and postoperative vein diameter. PMID- 7478716 TI - Urinary continence in Mullerian duct anomalies. AB - Mullerian duct congenital anomalies such as Rokitansky-Mayer syndrome, Urogenital Sinus, Vaginal Atresia and Cloacal Malformation are relatively uncommon (1-5% of born female newborns). The complexity of these malformations has taken great interest regarding mainly the surgical procedure available for correction of genital abnormalities. However, the problem of urinary incontinence is still underestimated, and continence is often a goal difficult to achieve. Authors report 11 cases of congenital anomalies of Mullerian duct differentiation which have been observed in the last decade in pediatric age with special regard to preoperatory diagnostic procedures, urinary continence valuation after surgical correction, therapy chances for continence, considering the psychological implication of this unresolved abnormality in everyday-lifetime. PMID- 7478717 TI - Wilson's disease in Albania. AB - There are 40 patients with Wilson's disease diagnosed in Albania; 24 were males and 16 females with an average age 18.8 +/- 8.47 years. The incidence rate was 1:75.000 and prevalence rate 1:148.000. According to clinical data, the Kayser Fleischer ring was found in 75% of the cases, the neurologic in 60% and mental disturbances in 27.5%. The liver was damaged in all the cases of acute and chronic hepatitis or liver cirrhosis. In 35% signs of hypersplenism were seen. Comparing these data with a control group of 60 alcoholic or viral cirrhosis with hypersplenism, it was found that hypersplenism is encountered in younger ages (p < 0.0001). The platelets were lower in WD than in the control group (p < 0.05), but leucocytes were lower in the control group (p < 0.001). Portal hypertension was found in 42.5% of the cases. Six of these (35.3%) were complicated by gastrointestinal hemorrhage. In two twins the disease was accompanied with another congenital anomaly: genu valgum bilateralis. Two other patients had hypertrophy of the parotis gland, six patients had signs of acute haemolysis, accompanied by nonspherocytic anemia, Coombs negative. PMID- 7478718 TI - A magnetic approach to AIDS. AB - Jacobson Resonance is the unified field equation yielding a frontier vision in magnetotherapy. The possible application to AIDS is considered. PMID- 7478719 TI - Epidemiological aspects, development and management of prevention in ophthalmology in relation to different ages. AB - In the ophthalmological field, it is not always easy to adopt a primary prevention. Much more useful would be a secondary prevention program towards the affections at birth or those later developed that, if diagnosed and treated early, grant a normal visual development. Differentiated prevention can be carried out, according to various ages: at the tender age congenital glaucoma and cataract, strabism, ametropia, anisometropia (possible cause of amblyopia) have to be considered; in the adult-senile age must be prevented damages due to hypertension, diabetes, thrombosis, as well as cataract, glaucoma, uveitis, kerato-conjunctivitis and retinal detachment. PMID- 7478720 TI - Pulmonary thromboembolism: current concepts and chronobiological perspectives. PMID- 7478721 TI - Giant sigmoid diverticulum. AB - The case of a giant diverticulum of the sigmoid colon in a 74 year-old woman is reported. The diverticulum had perforated into the urinary bladder and required a 3-phase operation with resection of the sigmoid colon and urinary bladder. Giant diverticulum is considered a rare complication of diverticulosis. Generally, it is a pseudo-diverticulum that grows due to a valve mechanism working in the neck of the diverticulum. The differential diagnosis lies mainly between intestinal duplication and pneumatosis cystoides. Surgical therapy yields favorable results. PMID- 7478722 TI - Penetrating war injuries of the heart. AB - Treatment of 12 penetrating heart war wounds were described. Ten wounded (83.3%) died on the spot or during transportation. Only two (16.7%) were brought to the clinic with signs of life, operated on and released from the hospital. The two wounded had not received any therapy before the arrival. The two cases described indicate the importance of a rapid transportation, adequate reanimation and the fastest possible cardiorrhaphy either by means of thoracotomy or medial sternotomy. PMID- 7478723 TI - Role of HPV-DNA typing in women with cytological diagnosis of squamous atypia. AB - The study was performed in 293 patients (mean age 36 years old). Pap tests, colposcopy and target biopsy were performed in all patients. Results showed 100 cases of cellular atypia, 53 cases of CIN (36 CIN I, 10 CIN II, 7 CIN III) and 140 normal cases. The last phase of the study consisted in the collection of exfoliated cervical cells which were then subjected to "Enzo Pathogen DNA Probe assay" using HPV-DNA 6/11, 16/18 and 31/33/51 probes. The aim of this study was to evaluate the relationship between HPV type and neoplastic evolution. The results were not sufficiently convincing to support proposing HPV/DNA typing as the sole diagnostic method in lesions of the last section of the female genital tract. PMID- 7478724 TI - The effect of aging and duration of disability on long term health outcomes following spinal cord injury. AB - The purpose of the study was to discover the independent and combined effects of age and duration of injury on selected long term health outcomes of 83 spinal cord injured (SCI) men (age range 21-79 years; duration of spinal cord injury 3 52 years). Specifically, the study examined a multivariate model which specified that age, duration, the interaction of age and duration, and level of lesion were related to the following health outcomes: pain, fatigue, functional independence, mobility, illness and symptomatology, perceived overall health, social support, life satisfaction, and economic stability. Age had significant main effects on three outcomes. With increasing age, the sample experienced more fatigue, decreased activity (due to pain), and more overall satisfaction with their lives. Duration of SCI showed significant main effects on two outcomes. As subjects lived longer with their SCI, they felt less financially secure and experienced more symptoms and illnesses. Significant interaction effects of age and duration of SCI were found for two outcomes. Older age combined with longer duration of SCI amplified subjects' perceptions of financial insecurity, and threats to health. The findings send a clear message to service-providers and policy makers about the added vulnerability of older disabled individuals, about the need for extra vigilance in health care issues, and about the necessity of a social economic safety net for already disadvantaged individuals. PMID- 7478726 TI - Depression in individuals with spinal cord injury: methodological issues. AB - The methodological problems in research related to depression in individuals with spinal cord injury (SCI) are examined. These include relation to normal emotional reactions following injury, the use of physical and vegetative symptoms in the diagnosis of depression, the utilisation of rating scales primarily designed for use in psychiatric populations, the heterogeneity of patients with SCI, the role of preexisting psychiatric morbidity, the selection of controls for comparison and the necessity for multivariate statistical approaches to analysis. The issues and possible solutions are discussed. PMID- 7478725 TI - Seminal somatostatin in men with spinal cord injury. AB - Somatostatin is present in high concentrations in human seminal plasma. Semen was obtained from men with spinal cord injury by vibratory-and/or electrostimulation. The seminal somatostatin concentration and sperm motility rate depended on the location of the spinal cord lesion. In men with lesions at or above T6 the seminal plasma concentration was significantly decreased compared to healthy men and to patients with lesions below T6 (P < 0.005). The latter group did not differ from healthy men. Men with spinal cord lesions below T6 had significantly decreased sperm motility rates (P = 0.09) compared to patients with lesions at or above T6. Seminal somatostatin concentrations were similar in vibratory- and electrostimulated ejaculates. The seminal somatostatin concentration, semen volume, sperm count, motility and morphology in each patient was not affected when repeated vibratory ejaculations were performed at home. PMID- 7478727 TI - Reinnervation of peripheral nerve segments implanted into the hemisected spinal cord estimated by transgenic mice. AB - We investigated how far Schwann cells, which are the peripheral nerve elements supporting axonal regrowth, penetrate into the hemisected recipient spinal cord. C57BL/6 mice, which carry carcinoembryonic antigen as transgene, were used for transplantation study. These CEA transgenic mice were syngenic to C57BL/6 mice except for the expression of human CEA DNA. In the syngenic transplantation study, C57BL/6 mice were transplanted with the sciatic nerve of CEA-transgenic mice to the hemisected spinal cord. Schwann cell migration into the recipient spinal cord was detected by the PCR method. Transplanted Schwann cells migrated into the recipient spinal cord both rostrally and caudally at a distance of 2 mm from the graft-host interface until 21 days after transplantation. At 28 days after transplantation, the Schwann cells migrated rostrally at a distance of 2 mm and caudally at a distance of 4 mm. C57BL/6-CEA sciatic nerve was transplanted to BALB/C mice as the allogenic transplant. CEA DNA was detected until 14 days after transplantation, but disappeared at 21 days. In addition, C57BL/6-CEA sciatic nerves were transplanted into Wistar rats to study xenogenic transplantation. The CEA band disappeared at 10 days after transplantation. In conclusion, by using CEA transgenic mice and the PCR method, we could evaluate the mobility of Schwann cells which are thought to play an important role in axonal regeneration. PMID- 7478728 TI - Analysis of arterial flowmotion in spinal cord injured and elderly subjects in an area at risk for the development of pressure sores. AB - Flowmotion (blood flow changes due to vasomotion) in the skin over a risk area for pressure sores, the sacrum, and a non-risk area, the gluteus muscle, was evaluated by laser Doppler fluxmetry during resting conditions and post-occlusive reactive hyperaemia (PRH) response. Measurements were made in healthy younger subjects and in two risk groups for pressure sores-spinal cord injured (SCI) and the elderly. The SCI were divided into two subgroups, one with and one without distinct flowmotion seen on the original recordings over the sacrum. The Prony spectral line estimation (PSLE) method was used to determine the power spectrum of the flowmotion activity. During the PRH, flowmotion frequencies were found in two separated bands, 5.4-6.6 cpm (cycles min-1) and 7.8-9.0 cpm. In the subgroup without distinct flowmotion, the PSLE method found flowmotion frequencies similar to the other groups. During the PRH, the flowmotion power pattern over the sacrum was similar within all groups, but the power was extremely low in one subgroup of SCI subjects. To conclude, flowmotion (vasomotion) was present in the two skin areas and increased during the PRH response, reaching a maximum within 50 s. This study shows that the flowmotion frequencies might be locally driven, whereas the power might be centrally mediated. Disturbances in microcirculatory flowmotion can be a part of the mechanisms leading to skin ischaemia and pressure sores. A new method has been developed for analysing differences in flowmotion behaviour such that statistical comparisons can be made. PMID- 7478729 TI - Biochemical analysis of heterotopic ossification in spinal cord injury patients. AB - Heterotopic ossification (HO) represents a frequent complication in spinal cord injury (SCI) patients. Samples of HO taken from SCI patients were studied and compared to normal bone. We used a procedure of bone particle fractionation (according to their degree of mineralisation) which allowed us to establish a profile reflecting the metabolic remodeling of bone and to analyse the organic matrix of the newly synthesised tissue. In paraplegic patients, we noted that there was a large increase of the proportion of a degree of calcified bone in the HO as we had previously observed in cortical as well as in cancellous bone of the same patients. Based on aminoacid analyses, we observed in the newly synthesised organic matrix of HO a decreased proportion of hydroxyprolyl residues resulting either from an alteration of the prolyl hydroxylation or from the presence of an excess of non-collagen polypeptides. These results are similar to those seen in sublesional bone of the SCI patients. This study demonstrates that HO is a newly formed bone which has a high rate of turnover as is seen in growing bone. This must be taken into account for the treatment of the patients. PMID- 7478730 TI - Measurement of body fat in individuals with tetraplegia: a comparison of eight clinical methods. AB - Spinal cord injury (SCI) is a unique condition that markedly alters body composition, raising the possibility of having undefined effects on the traditional assumptions for body composition. In order to determine appropriate methods for the analysis of body composition in this population, 12 subjects with tetraplegia were studied for absolute weight as fat and percent fat by the following methods; bioelectrical impedance (BIA), dual energy X-ray absorptiometry (DEXA), total body potassium (TBK), total body water (TBW), and four anthropometric methods: Durnin and Womersley (DUR), Jackson and Pollack (J and P), Sloan (SLN) and Steinkamp et al (STK). The eight methods were compared with the mean of all means (24.4 +/- 2.2% fat), which was assumed to be the best estimate of percent fat. Four methods: BIA, DEXA, TBW and STK were not significantly different, while TBK, DUR, J&P and SLN were significantly (P < 0.009) different from the mean of the means. Using only the non-significantly different methods, repeat computation revealed TBW to have the smallest difference from the mean (0.1 +/- 2.8%), and DEXA to have the strongest correlation with the line of identity (r = 0.96, P < 0.0001). In conclusion, BIA, DEXA, TBW, and STK are equally valuable for estimating fat in those with tetraplegia. PMID- 7478731 TI - The efficiency of walking of paraplegic patients using a reciprocating gait orthosis. AB - A number of orthoses have been designed to restore stance and walking in paraplegic subjects. Among them, the reciprocating gait orthosis (RGO) appears to be one which is most widely adopted. The great energetic cost of orthotically aided walking is the main factor limiting the walking capability of paraplegic subjects. The purpose of the present research was to give a complete biomechanical evaluation of RGO locomotion in order to provide data that can be used for further improvements of the orthosis. From the whole body kinematics the mechanical work done to move the orthosis was estimated; by measuring the actual metabolic cost of RGO locomotion the efficiency was evaluated as the ratio between mechanical work and energy expenditure. It is found that RGO locomotion is a very demanding task in terms both of energy expenditure and of the mechanical work required. Furthermore, it is evident that the major determinant of this poor efficiency of RGO locomotion is the enormous amount of work required to move the orthosis which, in turn, reflects its poor mechanical characteristics. To accomplish the demand of real mobility, devices for transferring work from the upper to the lower portion of the body must be substantially improved. This technical aspect must be parelleled by suitable training programmes especially designed to improve the aerobic power of paraplegic subjects and their body composition (ie less fat and more muscle). PMID- 7478732 TI - The role of percutaneous endoscopic gastrostomy in spinal cord injured patients. AB - Our experience with percutaneous endoscopic gastrostomy (PEG) in spinal cord injured patients is described. We have shown it to be a safe, comfortable and easily managed method of providing an adequate nutritional and fluid intake in patients requiring long-term tube feeding. It avoids the risks of intravenous feeding and the serious complications of oesophageal ulceration and stricture, which may occur with prolonged nasogastric intubation. PMID- 7478733 TI - Ventilatory thresholds during wheelchair exercise in individuals with spinal cord injuries. AB - The ventilatory thresholds of 30 male wheelchair athletes were determined from their respiratory and metabolic responses to a continuously progressive exercise protocol to peak oxygen uptake on a wheelchair ergometer. The peak oxygen uptake (peak VO2), ventilatory threshold in 1 min-1 (VTL), and ventilatory threshold expressed as a percentage of peak VO2 (VT%) were measured for all subjects. Statistical analyses of selected subsamples were used to note sport and functional ability level differences in these variables. Analysis of peak VO2 by functional classification (old International Stoke Mandeville Games Federation system; classes 1A-5, plus an amputee class) indicated that paraplegic (classes 2 5) and the amputee class athletes (2.48 1 min-1) were higher than tetraplegic (classes 1A-1C) athletes (0.95 1 min-1). The paraplegic and amputee classes were combined for a comparison of peak VO2 by sport which showed that track athletes (2.80 1 min-1) were higher than basketball players (2.41 1 min-1) who were higher than athletes from other sports (1.88 1 min-1). The VTL analyses demonstrated differences similar to the peak VO2 analyses. The VT% analyses, however, showed no sport differences, but the tetraplegic athletes had higher VT% values (87%) than the paraplegic plus amputee group (69%). PMID- 7478734 TI - Reversible anterior cord syndrome due to penetration of the spinal canal by pedicular screws. AB - The use of transpedicular screws in patients with disturbed posterior column anatomy is sometimes hazardous. We report a patient with a burst fracture at T7 who developed an anterior cord syndrome immediately after posterior instrumentation. Diagnosis of the anterior cord syndrome was ascertained by an emergency computerised tomogram which showed a misplaced screw compressing the cord from the anterolateral aspect of the spinal canal. Extraction of this screw and revision of the instrumentation resulted in speedy and complete resolution of the neurological deficit. The patient was mobilised within the first postoperative week and was able to work by the fourth postoperative month. The application of smooth contoured screws and blunt surgical instruments was found to be helpful because they are less likely to produce a cutting injury of neural tissue. We must further stress that transpedicular screws for the upper thoracic segments are to be employed very cautiously, if ever, to avoid severe neurological complications. PMID- 7478735 TI - Catheters and sounds: the history of bladder catheterisation. PMID- 7478736 TI - Soft tissue injuries associated with traumatic locked facets in the cervical spine. AB - The clinical, radiological and operative findings in 10 consecutive patients with cervical spine trauma presenting with locked facets (bilaterally in nine patients) are reported. The treatment was cervical traction until reduction of the locked facets was achieved, followed by anterior surgical decompression and ostheosynthesis. Intervertebral disc herniation was present in nine of the patients, and ruptured calcified posterior longitudinal ligament was present in the remaining patient. Four patients improved after treatment (none of these had clinically complete tetraplegia preoperatively), and three patients died. Associated soft tissue injuries producing spinal cord compression after reduction of cervical locked facets were verified in all of the patients of this series, suggesting that systematic surgical exploration should be done in this category of patients. PMID- 7478737 TI - Influence of electrical stimulation of the tibialis anterior muscle in paraplegic subjects. 1. Contractile properties. AB - In adult paraplegic subjects one tibialis anterior muscle received daily electrical stimulation for 4 weeks at twice motor threshold to determine changes of its contractile properties (this paper) and its morphological and histochemical profiles (following paper). Force, speed of contraction and fatigue resistance were assessed by percutaneous electrical stimulation of the muscle with torque measured about the ankle. Comparative contractile tests were performed on 51 normal adult subjects and new parameters for force measurement proposed, particularly where maximum voluntary contraction cannot be obtained. In paraplegic subjects before stimulation the tibialis anterior muscle showed evidence of disuse with decreased force, faster contraction and relaxation, and reduced fatigue resistance. The effects of two stimulus patterns were compared: 10 Hz, and 10 Hz with 100 Hz bursts. After stimulation contraction was slower, fatigue resistance increased and there was a tendency for force to increase. No differences occurred using the different stimulus patterns. Four weeks later fatigue resistance was partially maintained, while speed of contraction reverted to pre-stimulus levels. PMID- 7478738 TI - Superoxide radicals play important roles in the pathogenesis of spinal cord injury. AB - To investigated a possible role of superoxide dismutase (SOD) in the pathogenesis of spinal cord injury (SCI), the change in the circulatory status in an experimental SCI model of rats was examined. The blood pressure (BP) of the SCI group decreased markedly by compression or head-up-tilt (60 degrees). The extent of the BP decrease was diminished by the administration of along acting SOD (SM SOD) prior to injury. Compression, tilting and the administration of SM-SOD increased the sensitivity of resistant arteries to acetylcholine. SM-SOD treated rats had a significantly lower mortality than was found in untreated rats. These findings indicate that superoxide radicals may contribute to the pathogenesis of spinal cord injury and orthostatic hypotension caused by SCI. PMID- 7478739 TI - Pulmonary function testing in spinal cord injury: correlation with vital capacity. AB - Spinal cord injury (SCI) causes restrictive ventilatory changes, with reductions in vital capacity, functional residual capacity, and expiratory reserve volume. Vital capacity (VC) often is used as an indicator of overall pulmonary function in these patients. In an effort to determine the extent to which VC correlates with other pulmonary function tests, 52 patients with recent acute traumatic SCI underwent complete pulmonary function testing. Statistical relationships were determined between VC and nine other tests. VC was found to be significantly correlated with forced expiratory volume in 1 s, inspiratory capacity, expiratory reserve volume, functional residual capacity, residual volume (RV), total lung capacity (TLC), and RV/TLC ratio, but not with maximum positive expiratory pressure nor with maximum negative inspiratory pressure. The excellent correlations between vital capacity and nearly all of the other pulmonary function tests support the use of VC as a single global measure of overall ventilatory status in SCI patients. PMID- 7478740 TI - Ergonomy of paraplegic patients working with a reciprocating gait orthosis. AB - A reciprocating gait orthosis (RGO) is, among others, the most widely adopted device to restore the standing and walking capability of paraplegic patients. The aim of the present study was the evaluation of the energy demand (VO2), and cardiopulmonary load (HR and VE) imposed on the subject by different working tasks while sitting in a wheelchair or standing using a RGO. In addition, a comparison with the performance of normal subjects was also attempted. The RGO use allowed a dramatic improvement of patients' mobility and reach space in the workplace. A further advantage provided by the use of the RGO was represented by the increased mobility of the subjects with respect to the wheelchair confined situation. The energy demand and the cardiorespiratory load imposed on the subjects by the use of the RGO were not different from those observed both in the same subjects sitting in a wheelchair and in the controls. The energy demand slightly exceeded the values typical of light work and was, thus, compatible with the normal duration of a working day. On the other hand, the cardiac load corresponded to that typical of moderate activity, thus limiting the duration of the working task to 5-8 h. Based on the ergometry test, all of the working activities considered can be classified as aerobic activities, energy demand being under the ventilatory threshold. PMID- 7478741 TI - Association between muscle trauma and heterotopic ossification in spinal cord injured patients: reflections on their causal relationship and the diagnostic value of ultrasonography. AB - Paraplegic patients presenting with a subacute limitation of hip joint mobility were subjected to serial sonographic examinations. In four patients the initial sonographic study disclosed discontinuity with fluid collection in the psoas muscle, which was diagnostic of a traumatic muscle rupture. All four patients subsequently developed sonographic and radiographic evidence of heterotopic ossification. Our findings confirm that ultrasonography is an easy and inexpensive screening method for the early diagnosis of heterotopic ossification. The sonographic results obtained in these four paraplegic patients are indicative of a possible traumatic origin of heterotopic ossification around the hip. PMID- 7478742 TI - Traumatic spinal cord injuries in Istanbul, Turkey. An epidemiological study. AB - This is a retrospective study conducted in all of the hospitals of Istanbul to survey new patients with a traumatic spinal cord injury (SCI) in 1992. In that year 152 new traumatic SCI were identified. The estimated annual incidence was 21 per million population. The male/female ratio was 3/1. The mean age was 33, being 34 for male patients and 31 for female patients. 72% of all patients were under the age of forty. The major causes of SCI were falls (43%) and car accidents (41%), followed by being struck by an object (7%), gunshot injury (5%), stab injury (2%). Fifty patients (33%) were tetraplegic and 102 (67%) paraplegic. Regarding the tetraplegic patients the commonest level was C5, in those who were paraplegic L1. There were no cases at levels C1, T1 or T2. The commonest associated injury was head trauma, followed by fractures of an extremity (or extremities). Severe head trauma, as a major cause of death, may obscure the actual incidence of SCI in this study. Accidental falls (exceeding road accidents) were mostly due to falls from buildings and accidents on work premises. PMID- 7478744 TI - Reliability and validity of tympanic temperature measurement in persons with high spinal cord injuries. AB - Tympanic temperature measurements in the able bodied have been well studied and validated in previous investigations. This validation has not been studied in individuals with high spinal cord injuries where autonomic control is different above and below the level of the lesion, which makes the accuracy of this measurement for core body temperature questionable. In this correlational study we look at the reliability and validity of tympanic temperature measurement in individuals with high spinal cord injuries in comparison to oral and rectal temperature measures. The rectal measurement proved to be the most reliable with an r value of 0.975 overall. The oral was minimally better than the tympanic with r values of 0.88 and 0.86, respectively. The validity of the oral measure in predicting changes in the rectal temperature (gold standard) was slightly better than the tympanic with r values of 0.88 and 0.77, respectively. Both the oral and tympanic measures accurately reflected changes in the rectal (core) temperature. PMID- 7478743 TI - Morphological changes in the skin microlymphatics in recently injured paraplegic patients with ilio-femoral venous thrombosis. AB - Based on morphological features of the lymphatic microcirculation of the skin from healthy subjects, and from paraplegic patients who had no evidence of ilio femoral venous thrombosis (thromboembolic disease: TED), the leg terminal lymphatic vessels from skin biopsies of five male paraplegic patients with acute traumatic spinal cord lesions and with documented TED were studied. Paraplegic patients with TED had lymph vessels with a dilated lumen surrounded by a rarefacted perivascular connective tissue characterized by dissociation and disruption of collagen and elastic fibres. The lymphatic wall was generally attenuated and some open junctions and channels delimited by endothelial protrusions were observed. The venous outflow obstruction caused by deep venous thrombosis accompanied by the absence of ambulatory venous pressure in the paretic leg determines skin microlymphatic dilatation, lymph stasis and changes in the interstitial connective tissues. These alterations may be considered to be the morphological aspect of the dystrophic alterations seen in the skin of legs from paraplegic patients with TED. The results are discussed in view of the correct rehabilitative medical treatment necessary, and adequate prophylaxis of TED. PMID- 7478745 TI - Direct erosion of lumbar spine by an abdominal aortic aneurysm, resulting in paraparesis: unusual presentation. Case report. AB - Abdominal aortic aneurysm is a condition affecting nearly 4% of the elderly population. It has a potential for producing a wide range of symptoms, including abdominal pain and back pain. The latter is particularly difficult to interpret in patients with chronic rheumatological conditions, and delayed diagnosis may be associated with a poor outcome. We present a patient with rheumatoid arthritis and chronic low back pain, who developed bilateral leg weakness and hesitancy of micturition, due to an abdominal aortic aneurysm invading the spine. PMID- 7478746 TI - Latent tarsal tunnel syndrome with the provocation of flexor spasms in a paraplegic person. Case report. AB - A 71-year-old paraplegic woman presented with a complaint of insomnia caused by severe flexor spasms of the lower limbs at night, aggravated for a few hours by taking hot baths. Lidocaine injection of the tarsal tunnel diminished the flexor withdrawal reflex, which was easily elicited by light pinching or sustained compression over the tarsal tunnel, and strongly suggested the existence of the tarsal tunnel syndrome. Surgical decompression of the tarsal tunnel significantly reduced the flexor spasms both in terms of frequency and of duration. PMID- 7478747 TI - Solitary eosinophilic granuloma of the lumbar spine in an adult. Case report. AB - Eosinophilic granuloma of bone is the localised and most benign form of Langerhans-cell histiocytosis, previously known as histiocytosis X and is characterised by lytic lesions of one or more bones. It is a disease of children and adolescents, and very rarely affects adults. We report a 34-year-old patient with a solitary eosinophilic granuloma involving the fifth lumbar vertebra which produced sciatica. Operative treatment performed by a single extended posterior approach consisted of total resection of the tumour with hemicorporectomy, L4-5 and L5-S1 discectomy, interbody fusion with an iliac bone graft, and interpedicular fixation. Postoperatively, the symptoms resolved and the patient continues to do well at 2 1/2 years follow-up. PMID- 7478748 TI - Characteristics of accumulation of oral gram-positive bacteria on mucin conditioned glass surfaces in a model system. AB - Strains of Streptococcus, Actinomyces and Lactobacillus were grown on glass surfaces in semi-defined medium (pH 7.0) with mucin, at a dilution rate of D = 0.1 h-1, in a modified chemostat. The accumulation of cells followed four phases. In phase 1 (0-1 h), cells did not divide on the surfaces and adhesion accounted for rapid accumulation. Phase 2 (1-4 h) comprised adhesion and cell division, and accumulation slowed, cell number doubling times (Cdt) Streptococcus, 2.7 h to 8.6 h, Actinomyces, 2.3 h to 7.5 h and Lactobacillus, 3.6 h to 3.8 h. Cell division on surfaces accounted for accumulation in phase 3 (4 h to 12 h): Cdt Streptococcus, 1.7 h to 5.2 h, Actinomyces, 2.4 h to 7.5 h and Lactobacillus, 2.2 h to 7.2 h. The biofilm stabilized in Phase 4, Cdt 18.5 h to 90.2 h. The numbers (10(6) colony-forming units per cm2) of cells in stable biofilms were Streptococcus, 4.02 to 5.12, Actinomyces, 12.5 and 34.0 and Lactobacillus, 2.77. Accumulation increased (Cdt 0.9 h-2.7 h) when cells were exposed to glucose excess or high dilution rates and phase 2 of accumulation did not occur. PMID- 7478750 TI - The influence of saliva on interbacterial adherence. AB - The mechanism of bacterial adherence to the mat of bacteria in preformed dental plaque is not well defined. This study measured the influence of saliva on the adherence of bacteria in suspension to a continuous bacterial surface in vitro. Twenty different pairs of bacteria were tested, consisting of Streptococcus spp., Haemophilus spp. and Actinomyces spp. The species were chosen based on the parameters of coaggregation, and salivary agglutination. The results were expressed as bacteria that adhered per mm2 of bacterial surface. When both the surface bacteria and the bacteria in suspension agglutinated in saliva, interbacterial adherence was increased 2.5-fold when a salivary coating was placed on the surface. When one or both of the bacteria did not agglutinate in saliva, interbacterial adherence was increased only slightly when the surface was saliva-coated. The results suggested that salivary-mediated adherence is significant to plaque formation once the tooth surface becomes covered with bacteria. Thus bacteria that are capable of agglutinating in saliva may have a distinct advantage in colonization of the plaque surface. PMID- 7478749 TI - Surface-associated properties of Actinomyces strains and their potential relation to pathogenesis. AB - Twenty-nine strains from the Actinomyces species were tested for a range of surface properties. Results show considerable heterogeneity both between different species and within some of the species, especially Actinomyces naeslundii. Two commonly used A. naeslundii strains, T14V and ATCC 12104, fell within the low (salivary aggregation and collagen binding by T14V), moderate (surface charge and haemagglutination) or high range of values (hydrophobicity, saliva-coated hydroxyapatite adhesion, polystyrene binding by T14V, fibrinogen binding by T14V and collagen binding by A. naeslundii ATCC 12104). Both strains adhered well to saliva-coated hydroxyapatite; T14V bound the highest amount of fibrinogen, ATCC 12104 had the highest number of cells bound to collagen and T14V was not bound at all. The heterogeneity of these characteristics highlights the need to include a range of strains of Actinomyces in studies on their pathogenicity. Statistical correlations were found between a number of properties, for example saliva-coated hydroxyapatite adhesion and hydrophobicity, and between haemagglutination and hydrophobicity. PMID- 7478751 TI - Growth-inhibitory effect of pyrophosphate on oral bacteria. AB - The purpose of this investigation was to determine whether pyrophosphate, the anticalculus component of tartar-control dentifrices, exerts antimicrobial activity against oral bacteria commonly found in supragingival plaque. Minimal inhibitory concentrations of pyrophosphate were determined for Streptococcus sanguis, Streptococcus mutans (serotype c), Actinomyces viscosus and Actinomyces naeslundii. All of the bacteria tested were susceptible to pyrophosphate with identical minimal inhibitory concentrations of 0.67% wt/vol (25 mM). Bactericidal kinetics assays revealed that both S. mutans and A. viscosus were killed by pyrophosphate, with the latter being considerably more susceptible. The mechanism of killing was not due to high ionic strength, as comparable controls showed no loss in numbers of viable cells. Brief exposure (two 5-min incubations) of S. mutans to pyrophosphate and sodium dodecyl sulfate caused pronounced inhibition of growth over the 24-h test period. Under the constraints of the conditions used, these studies indicate that pyrophosphate and sodium dodecyl sulfate can substantially inhibit the growth of oral bacteria. These compounds may affect the oral microflora of patients who routinely use tartar-control dentifrices and mouthrinses. PMID- 7478752 TI - Catabolite modification of acid tolerance of Streptococcus mutans GS-5. AB - The acid tolerance of Streptococcus mutans GS-5 in standard pH-drop assays was found to be affected by the sugar used in the assay and also by the sugar used for growth of the organism. For example, acid tolerance was lower when galactose was used as catabolite than when glucose was used, apparently because galactose/proton symport brought protons extruded by the F-ATPase back into the cell and thus reduced delta pH across the cell membrane. The acid tolerance of glycolysis was related directly to the capacities of the cells to produce acid glycolytically, or probably more correctly, to their capacities to produce adenosine triphosphate but not to acid tolerance of phosphotransferase systems for sugar uptake. Thus, glycolytic acid tolerance of S. mutans depends not only on environmental factors such as potassium or magnesium levels but also on the specific catabolites the organism is metabolizing or to which it has become metabolically adapted. PMID- 7478753 TI - Effective method for discriminating between oral bacterial and human alkaline phosphatase activity. AB - Alkaline phosphatase (ALPase) activity was quantitatively compared in various kinds of oral bacteria. High ALPase activity was detected in 3 species of periodontal bacteria, Porphyromonas gingivalis, Prevotella intermedia and Capnocytophaga sputigena. The ALPase activity detected in these bacteria was almost completely inhibited in the presence of 1% sodium dodecyl sulfate (SDS). By contrast, the activity of mammalian ALPase isoenzymes was not inhibited at all even in the presence of 1% SDS. These results indicate that the ALPase assay in combination with 1% SDS can identify the origin of ALPase detected in gingival crevicular fluid as being from bacteria or from a host response. Clinical examination with adult periodontitis revealed that ALPase activity in gingival crevicular fluid from the patients consisted of a combination of SDS-sensitive and SDS-resistant activities. These findings indicate that ALPase activity detected in gingival crevicular fluid originates not only from bacteria but also from a host response. PMID- 7478754 TI - Oral microflora of the ferret at the gingival sulcus and mucosa membrane in relation to ligature-induced periodontitis. AB - The aim of this study was to describe, in the domestic ferret, the composition of the normal gingival sulcus and the near related mucosa microflora and the shift in the composition during an acute periodontal breakdown induced by the placement of ligatures. Five animals were used. At the start of the experiment (day 0) and before the placement of the ligatures, 2 microbiological samples were obtained from the gingival sulcus and the related mucosa at the mid-buccal surface of the right upper P4. The same sites were sampled on the left side, serving as control sites. On day 28, new microbiological samples were taken from the same 4 sites. The samples were analyzed with darkfield microscopy and with viable counts. In the darkfield analysis, on day 0, the predominant bacterial morphotype was cocci, ranging from 88-92%. The viable counts showed that nearly 100% of the total cultivable flora was composed of facultative anaerobic gram-negative and gram positive rods. Pasteurella spp., Corynebacterium spp. and Rothia spp. were the major components of these bacterial groups. No anaerobic bacteria were detected. On day 28, cocci decreased to approximately 75% in the experimental gingival sulcus sample, and filaments and rods increased significantly. Spirochetes remained largely unchanged during the experiment. The composition of the experimental site samples changed. There was a decrease in the mean numbers of facultative anaerobic gram-positive rods, and approximately one third of the microflora comprised anaerobic gram-negative rods, mainly Porphyromonas gingivalis and Fusobacterium spp. Although the composition of the microflora of the ferret presented few similarities to that described in dogs and monkeys, the anaerobic portion of the gingival sulcus and oral mucosa microflora related to ligature-induced periodontitis presented microorganisms observed in experimental periodontitis both in dogs and monkeys as well as in human periodontal diseases. PMID- 7478755 TI - Use of the Syrian golden hamster for the induction of intraoral abscesses by sutures contaminated with human subgingival plaque. AB - Analysis of normal oral flora in 150 cheek pouches of hamsters (Mesocricetus auratus) defined the microbial working environment and demonstrated the absence of human oral black-pigmented bacteria. Silk sutures saturated with Porphyromonas gingivalis, Prevotella intermedia or subgingival plaque were used to close wounds made in hamster's cheek pouches. Abscesses were formed when sutures had solitary P. gingivalis or other bacteria mixed with P. gingivalis or when P. intermedia was mixed with other bacteria besides P. gingivalis. A concentration of black pigmented bacteria emanating from 3 x 10(5) colony-forming units/inoculum was required for abscess formation. Six abscesses (14.3%) were developed in association with the presence of other odontopathic bacteria, primarily Fusobacterium nucleatum and Actinomyces viscosus. The hamster cheek pouch with iatrogenic wounds closed with plaque-impregnated sutures is a novel and effective model to study the pathology of wound infections and virulence of human subgingival organisms. PMID- 7478756 TI - The effects of human immunodeficiency virus infection on macrophage phagocytosis of Candida. AB - The effects of infection by human immunodeficiency virus (HIV) on phagocytic function towards Candida pseudotropicalis were tested using monocytotropic and nonmonocytotropic HIV strains on peripheral blood monocytes. There were no differences in either quantitative phagocytic activity or efficacy between the two infected and control mock-infected monocytes. Immunofluorescence for cytoplasmic p24 antigen and in situ hybridization for detection of HIV-specific RNA sequences revealed that very few cells (10(-4)) exhibit productive infection. Occasional cells with active infection as defined by microscopically detectable fluorescence labelling contained phagocytized yeasts. PMID- 7478757 TI - Innervation of recurrent aphthous ulcers. AB - Specimens from nonkeratinized oral mucosa were obtained from diseased and clinically healthy mucosa from 7 patients with minor recurrent aphthous ulcers. The innervation of the specimens was visualized using antibodies to neuron specific intermediate cytoskeletal neurofilament fiber, the cytoplasmic protein gene product 9.5 and a 38 kDa integral membrane protein of synaptic vesicles applied in avidin-biotin-peroxidase staining. Mapping with these 3 antibodies revealed dense and basically similar pattern of innervation in the specimens of the clinically healthy oral mucosa. In recurrent aphthous ulcers, all 3 general markers disclosed peripheral nerve fibers also in the lesions, apart from the necrotic area, among the inflammatory cells without signs of retraction from the diseased area. Synaptophysin staining suggested that these peripheral nerve fibers in the inflammatory areas still contained synaptic vesicles. Accordingly, they were shown to contain substance P and calcitonin gene-related peptide, which are known to be released upon stimulation of the nerve and can exert potent paracrine actions, possibly on the local inflammatory cells as suggested by a close spatial relationship between neuropeptide-containing nerves and inflammatory cells. PMID- 7478759 TI - The antimicrobial activity of essential oils and essential oil components towards oral bacteria. AB - A method for reproducibly determining minimal inhibitory concentrations and minimal bactericidal concentrations of plant extracts towards fastidiously and facultatively anaerobic oral bacteria, predicated upon measurements of optical densities in microtitre plate wells, was devised. The antimicrobial properties of some botanical oils were surveyed; of these, Australian tea tree oil, peppermint oil, and sage oil proved to be the most potent essential oils, whereas thymol and eugenol were potent essential oil components. PMID- 7478758 TI - In vitro modulation of oral bacterial adhesion to saliva-coated hydroxyapatite beads by milk casein derivatives. AB - Bovine caseinate, derivatives of its glycosylated moiety [caseinoglycomacropeptide (CGP)], and caseinophosphopeptides were evaluated as inhibitors of adhesion of oral bacteria to saliva-coated hydroxyapatite beads (S HA). All milk casein-derived components behaved as potent inhibitors of Streptococcus sanguis OMZ 9 and Streptococcus sobrinus OMZ 176 adhesion to S-HA, whereas neither bovine serum albumin nor polyethyleneglycol were able to interfere with the adhesion of these strains. By contrast, none of the molecular species tested was able to inhibit the attachment of Actinomyces viscosus Ny 1 to S-HA. On the other hand, casein derivatives were shown to displace human serum albumin from S-HA beads. They were also able to bind to the bacterial cell surface of all strains examined. Collectively, these findings suggest that interactions between acidic casein-derived milk components and the biological surfaces involved in bacterial adhesion to S-HA result in an inhibitory effect that is selective for the oral streptococci examined. PMID- 7478760 TI - Biochemical change exhibited by oral streptococci resulting from laboratory subculturing. AB - The intent of this study was to assess the effects of continued laboratory subculturing on selected biochemical properties of oral streptococci freshly isolated from dental plaque. Six fresh isolates (3 Streptococcus mutans and 3 non mutans) and 2 laboratory strains were subcultured daily for a total of 225 transfers, and cells were harvested every 75 transfers from duplicate batch cultures grown with glucose at a constant pH. Eleven biochemical properties were assayed with cells, membranes and cell-free extracts and the results subjected to statistical analysis for differences between the duplicate cultures and the various subcultures. In addition, the activity of 19 hydrolytic enzymes was assayed with the semiquantitative apiZYM system (Analytab products). The activity of zero-time samples varied by as much as 241-fold for a single property with particularly low activity for EIIglc of the phosphoenolpyruvate phosphotransferase system and cell-associated extracellular polysaccharide synthesis. The 3 S. mutans fresh isolates had higher activity in 8 of the 11 assays compared with the 3 non-mutans strains, with extracellular polysaccharide synthesis the most significant trait. Statistical analysis of the 2816 assays of the 11 traits for the 8 test strains at the 4 selected time intervals revealed considerable change in the activity of the test parameters. The most notable changes in the S. mutans strains over the 225 subcultures were significant increases in glycolytic activity and decreases in hydrophobicity and extracellular polysaccharide synthesis activity. Of the measured properties, lactate dehydrogenase and cell-associated extracellular polysaccharide synthesis activity were the most stable and H+/ATPase activity was the most variable.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7478761 TI - The role of the succinate pathway in sorbitol fermentation by oral Actinomyces viscosus and Actinomyces naeslundii. AB - The sorbitol fermentation by Actinomyces viscosus and Actinomyces naeslundii was studied with washed sorbitol-grown cells. The fermentation was followed by titration of acids produced at pH 7.0 under anaerobic conditions. Metabolic end products and intracellular levels of NAD, NADH and glycolytic intermediates during the fermentation were also analyzed. Cell extracts were examined for certain enzyme activities. Bicarbonate was required for acid production from sorbitol and from a mixture of glucose and sorbitol. Malate and fumarate could also support the acid production of A. viscosus. The main end-products were succinate and lactate but not ethanol. Cell extracts showed no activities of alcohol and aldehyde dehydrogenases, but they had activities of malate dehydrogenase and fumarate reductase. In the absence of bicarbonate, malate or fumarate, the intracellular NADH/NAD ratio increased and the levels of 3- and 2 phosphoglycerate and phosphoenolpyruvate decreased. The results indicate that oral sorbitol-fermenting actinomyces lack the ethanol pathway that can contribute to NADH oxidation. To maintain intracellular redox balance during anaerobic sorbitol fermentation, these bacteria can oxidize surplus NADH through a succinate pathway. PMID- 7478763 TI - The in vitro proteolytic and saccharolytic activity of Candida species cultured in human saliva. AB - The proteolytic and saccharolytic activity of 4 Candida species was investigated in batch cultures of pooled, human mixed saliva supplemented with glucose. All the Candida species investigated (Candida albicans, Candida tropicalis, Candida glabrata and Candida krusei) demonstrated a marked growth in saliva with a concomitant reduction in pH from about 7.5 to 3.3, within 72 h. Isotachophoretic analysis of the culture supernatant revealed the presence of a variety of acid anions of which pyruvate and acetate were the most abundant. Proteolysis of salivary components, evaluated by a biochemical assay and sodium dodecyl sulfate polyacrylamide gel electrophoresis, was exhibited by all 4 Candida species, although there was inter-species variation. Despite the similarity in growth rates, C. tropicalis and C. krusei demonstrated greater proteolytic activity than C. albicans and C. glabrata. Neither candidal growth nor proteolysis was observed in glucose-free control saliva samples. In contrast, the degree of saccharolytic and proteolytic activity of a single isolate of C. albicans in glucose supplemented parotid saliva appeared to be relatively weak compared with mixed saliva. As the oral cavity provides ideal low pH niches periodically supplemented with dietary carbohydrates, the acidic proteinases of Candida species may play a role in the pathogenesis of oral candidiasis. PMID- 7478762 TI - Effect of proteolytic enzymes on the lysis and growth of oral bacteria. AB - Previous studies have shown increased levels of proteolytic enzymes in affected periodontal sites. The aim of the present investigation was to evaluate the effect of proteolytic environments on the lysis and growth of selected oral bacteria associated with either healthy or diseased periodontal sites. The effect of trypsin, chymotrypsin and proteinase K on cell lysis was determined following incubation with bacteria, whereas the effect of the same proteolytic enzymes on bacterial growth was tested using a disc-plate technique. Overall, gram-positive bacteria appeared to be more resistant to lysis than gram-negative bacteria. The most susceptible bacteria were Actinomyces spp., Eubacterium saburreum, Prevotella intermedia, Capnocytophaga ochracea, Fusobacterium nucleatum, Prevotella loescheii, Treponema denticola and Actinobacillus actinomycetemcomitans. The disc-plate procedure indicated that the growth of Actinomyces spp., E. saburreum, C. ochracea, P. intermedia, P. loescheii, Porphyromonas gingivalis and T. denticola were the most affected, more particularly by chymotrypsin and proteinase K. Interestingly, the growth of F. nucleatum was rather stimulated by proteolytic enzymes. The observations reported in this investigation indicate that specific and general proteolytic activities have the ability to lyse some oral bacterial species and to interfere with their growth. It is suggested that such effects could represent new mechanisms by which the bacterial ecology of subgingival sites may be affected. PMID- 7478764 TI - The effect of indigenous bacterial populations on buccal epithelial cells on subsequent microbial adhesion in vitro. AB - Despite the numerous investigations on the adhesion of microorganisms to buccal epithelial cells, it is difficult to correlate different results, as disparate adherence values have been reported for a given organism. As one reason for this disparity may be the indigenous or natural bacterial populations on human buccal epithelial cells, the effect of the latter on subsequent microbial adherence in vitro was examined. There was a highly significant correlation between the degree of natural bacterial colonization on pooled buccal epithelial cells from 8 healthy donors and the adhesion of a single isolate each of Streptococcus mitis, Escherichia coli and Actinomyces naeslundii. However, no such relationship could be established for Candida albicans, Streptococcus milleri and another isolate of Streptococcus mitis. As in previous studies, variation in adherence values was found, both between samples from different donors, and from the same donor over time, but to a far lesser degree in pooled samples from different donors. These results imply that natural bacterial populations on buccal epithelial cells may affect the adhesion values derived from laboratory experimentation, and hence such data should be interpreted with caution. PMID- 7478765 TI - Host-related genotypic heterogeneity of Porphyromonas gingivalis strains in the beagle dog. AB - The present investigation explored the genotypic heterogeneity of Porphyromonas gingivalis using restriction endonuclease analysis and ribotyping of 64 P. gingivalis isolates, recovered from the periodontal pockets of 3 beagle dogs, 2 of which were reared together. The isolates originated from both healthy and periodontal disease affected sites and thereby enabled the study of bacterial genotype with respect to (i) individual host, (ii) ecological niche (site within host) and (iii) level of periodontal health. Whole genomic DNA was extracted from each isolate and digested by the restriction endonuclease KpnI. Digestion fragments were separated by electrophoresis and transferred onto nylon membranes. The blots were hybridized with a digoxigenin-labeled 16S rDNA probe, and hybridization bands were detected using an anti-digoxigenin antibody conjugated with alkaline phosphatase and enhanced chemiluminescence. Fourteen genomic fingerprints and 13 ribotypes were observed among the 64 isolates. As many as 8 distinct fingerprints were detected within a single host and up to 4 fingerprints within a single periodontal pocket. The dogs reared together shared 2 common clonal types but also exhibited clonal types unique to each dog. No clear association between clonal type and periodontal health status could be made. The results revealed an extensive intra-host genotypic heterogeneity of P. gingivalis strains in the beagle dog and indicated that ribotyping was a sensitive method for differentiating clonal types within species. PMID- 7478766 TI - Mitsuokella dentalis in human periodontitis. AB - The occurrence of Mitsuokella dentalis in periodontitis was determined by culture and DNA probe detection. Subgingival paper-point samples from 480 periodontitis patients were transported in VMGA III, plated onto brucella agar with 5% sheep blood and incubated anaerobically for 7 days. Presumptive identification was based on a colony morphology resembling a water drop and biochemical characteristics. DNA probe detection was performed on paper-point samples using a digoxigenin-labeled cellular M. dentalis DNA probe in a dot-blot assay. Culture and DNA probe identified M. dentalis in 18.1% and in 80.7% of the study patients, respectively. M. dentalis isolates produced phosphatases, galactosidase, glucosidase and acetylglucosaminidase and showed high in vitro sensitivity to metronidazole. This study revealed that M. dentalis is a constituent of the pathogenic microbiota in human periodontitis. The periodontopathic potential of the organism is unknown. PMID- 7478768 TI - The time for political advocacy for children is now. PMID- 7478767 TI - The tet(Q) gene in bacteria isolated from patients with refractory periodontal disease. AB - Twenty-two tetracycline-resistant (tetr) anaerobic and facultative anaerobic bacteria isolated from periodontal pockets of 12 patients with refractory periodontitis were examined for the presence of the Tet Q determinant by DNA-DNA hybridization. Dot blots of bacterial DNA were tested with an intragenic digoxigenin-labelled tet(Q) probe consisting of a 1.45 kb EcoRI/PvuII fragment from plasmid pNFD13-2. Southern blots of chromosomal DNA digested with the restriction enzyme EcoRI were also examined. The tet(Q) probe hybridized with DNA from 8 of the 22 tetr strains, including 2 Prevotella intermedia strains and one strain each of Prevotella nigrescens, Prevotella loescheii, Prevotella veroralis and Prevotella melaninogenica. The tetr strains of Mitsuokella dentalis and Capnocytophaga ochracea also hybridized with the probe. The lack of discernible plasmid DNA in all the probe-positive isolates suggests that these tetracycline resistance genes were chromosomally encoded. The probe hybridized with a different size fragment in all the isolates. This study extends the number of species that carry the tet(Q) gene to include several outside the genera Prevotella and Bacteroides. PMID- 7478769 TI - Political advocacy from a pediatrician's perspective. PMID- 7478770 TI - The practice of political medicine. PMID- 7478771 TI - Influencing the political process through coalitions: the Michigan Council for Maternal and Child Health. AB - Building a coalition with others is an effective tool for increasing influence at the state level of the political process. It allows for the hiring of a staff who are able to maintain a constant presence in the ever-changing state political arena, which individual physicians and other caregivers simply cannot do. It allows for the development of increased sophistication among its members, which likewise increases the ability to affect the political process. It should be done with a philosophical set of standards that preserves its integrity and focuses on its goals, which must be carefully delineated from the inception. Coalitions are an effective way to deal with public issues of maternal and child health. PMID- 7478772 TI - Pediatricians: a special interest group for children. PMID- 7478773 TI - The use of media to impact on legislation. AB - There clearly are a large number of opportunities to use the media to influence legislation and legislative initiatives. There are a number of examples of legislation or laws that now exist as a result of pediatricians using the media effectively. National requirements for child auto restraints, Consumer Product Safety Commission regulations on children's toys, safety caps on medications, bicycle helmet laws, and increasing restrictions on guns can all be traced, in part, to the timely and appropriate use of the media. North Carolina safe gun laws to protect children resulted from pediatricians using some unfortunate and tragic incidents to dramatize the need for gun control. Highlighting the inappropriate care for children in emergency situations in New Jersey contributed to the movement to enact pediatric emergency care guidelines for that state. Finally, the effective use of the media by the AAP has brought a number of serious, child-related issues to the legislative forefront resulting in improved conditions for children. It is not easy, though. To use the media effectively takes persistence, thoughtful attention to developing relationships with the professionals in the media, and careful follow-up. It's important that pediatricians speak out on behalf of children and children's needs. The media, when used properly and knowledgeably, can be a powerful ally and contributor to a better outcome. PMID- 7478774 TI - Getting involved with the Federal Government: six questions and answers. PMID- 7478775 TI - Characteristics of near-fatal asthma in childhood. AB - As part of the South Australian asthma mortality survey, we examined 30 cases of near-fatal asthma attacks in children under 15 years of age who were seen over a 3-year period from May 1988 to June 1991. Subjects presented with asthma and either respiratory arrest, PaCO2 above 50 mm Hg, and/or an altered state of consciousness or inability to speak on presentation at a metropolitan Adelaide teaching hospital. A standardized interview and questionnaire was completed with subjects/parents and medical practitioners. Data were reviewed by the assessment panel which made collective judgments based on predetermined criteria. Seventeen patients (57%) were male, 20% were less than 7 years of age, and the majority (53%) were aged between 12 and 15 years. The majority (83%) had severe asthma and only one case (3.3%) had mild asthma. Half of the subjects were waking every night due to asthma and 79% had significant exercise limitation. A quarter of the subjects had a previous ICU admission and 70% had a hospital admission in the last 12 months. Primary care was carried out by a general practitioner in 57% of cases, and 70% of subjects had a crisis plan. Only 46% of those older than 7 years of age had ever used a peak-flow meter. Eighty percent of subjects or their families had high denial scores, and in 73% of cases psychosocial factors were considered to be significant. Eighty percent of cases experienced acute progressive respiratory distress, and 63% of cases delayed seeking medical care.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7478776 TI - Aerosolized furosemide in wheezy infants: a negative report. AB - We investigated the effects of 10 mg aerosolized furosemide on clinical score in 28 acutely wheezing infants (Part A) and in a second group of 20 intermittently wheezing babies on airway resistance and functional residual capacity during a symptomfree period (Part B), using a double-blind, placebo-controlled design. In both parts of the study no therapeutic effects were observed during and following aerosol inhalation of 10 mg furosemide. PMID- 7478777 TI - Late pulmonary function following repair of tracheoesophageal fistula or esophageal atresia. AB - Although tracheoesophageal fistula and esophageal atresia (TEF-EA) are surgically correctable, late respiratory complications have been reported. We administered a respiratory and gastrointestinal symptom questionnaire and performed standard pulmonary function tests (PF T's) and methacholine challenge testing on an unselected group of 25 subjects with TEF-EA who underwent surgery at our institution between 1963 and 1985. Results were compared to predicted normals, as well as 10 sibling controls. While the mean values of lung function test results for the TEF-EA group were within the normal range, they were significantly different from their siblings. Thirteen of the 25 TEF-EA group (52%), but none (0%) of the controls, had abnormal pulmonary function. This was classified as restrictive in 9 (36%), obstructive in 3 (12%), and mixed in 1. In addition, airway hyperreactivity [defined as a positive methacholine challenge (PC20 < or = 8 mg/mL)], was found in 6 of 18 TEF-EA subjects and 4 of the 9 controls who were evaluated. Comparison of TEF-EA subjects with normal and abnormal PFTs showed no difference in the incidence of tracheomalacia, esophageal strictures or dilatation, recurrent pneumonias, or gastroesophageal reflux. The respiratory symptom score in the subjects and controls was similar, and did not correlate with abnormal pulmonary function. The cause of the pulmonary function abnormalities remains unexplained. PMID- 7478778 TI - Inhaled nitric oxide improves oxygenation in piglets with meconium aspiration. AB - We hypothesized that nitric oxide (NO) inhalation in a model of meconium aspiration in newborn piglets would decrease pulmonary vascular resistance. Seven neonatal piglets were obtained at less than 48 hr of age and instrumented under fentanyl anesthesia. Inhaled NO (40 parts per million) was administered during normoxia and again after hypoxia was induced by reducing FiO2 to 0.13. During normoxia NO inhalation caused a fall in pulmonary artery pressure from a mean of 3.15 (SD 0.8) kPa to 2.84 (SD 0.7) kPa (P < 0.01). Hypoxia (mean arterial O2 saturation 35%) increased PA pressures to a mean of 5.4 (SD 1.6) kPa and NO administration during hypoxia decreased PA pressures to 3.6 (SD 1.2) kPa (P < 0.001). In order to determine the effects of NO in a model of meconium aspiration, 6 to 7 mL/kg of 20% human meconium in normal saline was instilled into the trachea. This procedure induced hypoxemia (mean SaO2 43.4%, SD 19), respiratory acidosis, (mean PaCO2 12.1 kPa, SD 0.5; mean pH 7.04, SD 0.03), and pulmonary arterial hypertension (mean pulmonary artery pressure 6.0 kPa, SD 1.3) despite ventilation with 90% oxygen. Inhaled NO was then administered in concentrations of 5, 10, 20, 30, 40, 60, and 80 parts per million in random order according to a Latin square design.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7478779 TI - Effect of chestwall distortion on the measurement of pulmonary mechanics in preterm infants. AB - The purpose of this study was to determine the effect of chestwall distortion (CWD) on the measurement of pulmonary compliance (CL) and resistance (RL). Inductance plethysmography was used in 15 preterm infants to determine CWD as total compartmental displacement ratio (TCDR) and out of phase movement between ribcage and abdomen as phase shift (PS). Flow was measured by pneumotachography and esophageal pressure change (Pe) with a water-filled catheter. CL and RL were calculated by linear regression analysis. Seven infants (mean +/- SD: BW, 1,484 +/- 186 g, GA 32.4 +/- 2.2 weeks, age 8.7 +/- 4.7 days) had a breathing pattern characterized by episodes with a high degree of CWD, followed by periods with minimal CWD (distortion group). In this group lung function measurements were analyzed separately during periods with and without CWD. The remaining 8 infants (BW, 1,244 +/- 233 g, GA 30.4 +/- 2.4 weeks, age 7.4 +/- 3.1 days) always breathed with minimal CWD, and the measurements in this group (non-distortion group) were used as a reference for the values obtained in the distortion group. Measurements of TCDR, PS, CL, RL, and tidal volume (VT) obtained in the absence of CWD were not significantly different between distortion and non-distortion groups. The measurements obtained in the presence of CWD showed a significantly higher TCDR and PS, but CL and RL were not significantly different from the CL and RL measurements obtained in the distortion and non-distortion groups in the absence of CWD. The only significant effect of CWD was a reduction in VT.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7478780 TI - Sensitivity and specificity of oropharyngeal suction versus bronchoalveolar lavage in identifying respiratory tract pathogens in children with chronic pulmonary infection. AB - The purpose of this study was to compare the pathogens obtained by oropharyngeal suction (OPS) and bronchoalveolar lavage (BAL) in children with chronic pulmonary infections. Forty-four children (mean age of 6.1 years; range 4 months to 15 years) were included in the study (27 children with recurrent localized lung infection, 5 with bronchiectasis, 5 with cystic fibrosis, 2 with foreign body aspiration, 2 with bronchiolitis obliterans, and 3 with recurrent episodes of cough and shortness of breath). In 27 out of 44 BAL samples (61%) bacterial cultures were positive. The sensitivity of OPS in detecting the same BAL pathogen was 89% (24/27 samples), the specificity was 94% (16/17 samples), and the predictive value was 91% (40/44 samples). Hemophilus influenzae beta-lactamase negative was the main organism recovered from BAL in non-cystic fibrosis patients with recurrent or persistent pneumonia. We conclude that OPS is a simple and efficient noninvasive procedure which may be helpful in the diagnosis and treatment of recurrent or chronic pulmonary infection. PMID- 7478781 TI - Detection of Pseudomonas (Burkholderia) cepacia using PCR. AB - Pseudomonas cepacia colonization of the lung is associated with increased morbidity and mortality for cystic fibrosis (CF) patients. The lack of a sensitive detection method for Pseudomonas cepacia in CF sputum has resulted in controversy regarding its epidemiology. We designed a PCR method to detect P. cepacia using P. cepacia 16 S rRNA sequences as the amplification target region. The PCR amplification with purified DNA as template yielded the expected 209-bp products from P. cepacia, but not from related Pseudomonas species of medical importance or other bacteria which have been reported to colonize CF patients. In serial dilution experiments as few as 10(2) P. cepacia CFU were detectable. When sputum samples from three CF patients chronically colonized with P. cepacia and P. aeruginosa were analyzed, P. cepacia was detected in all three specimens by PCR, but only in two when selective culture was performed. Our data support the potential role of PCR technology in the rapid, sensitive, and definitive detection of P. cepacia in CF sputum samples, even in the context of concomitant P. aeruginosa colonization. PMID- 7478782 TI - Comparison of effects of an intrapulmonary percussive ventilator to standard aerosol and chest physiotherapy in treatment of cystic fibrosis. AB - Impaired mucociliary clearance due to defective ion and water transport and the effects of chronic airway infections lead to stasis of secretions and progressive pulmonary damage in patients with cystic fibrosis (CF). Methods to improve removal of tenacious lung secretions in CF patients contribute to slowing the decline in respiratory function. We have evaluated an intrapulmonary percussive ventilator (IPV), which is a device designed to enhance airway clearance and preserve lung function. A previous pilot study by us had determined that the device was acceptable to patients and is safe. We undertook a 6 month parallel comparative trial of the IPV versus standard, manual chest physiotherapy in 16 CF children and adults. No significant differences in spirometric measures, numbers of hospitalizations, use of oral or IV antibiotics, or anthropometric measurements were detected between the standard aerosol/chest physiotherapy group and the IPV group over the duration of the trial. Patient acceptance, as determined by participant survey, was good. The device appeared to be safe and durable. It was concluded that the IPV is as effective as standard aerosol and chest physiotherapy in preserving lung function and anthropometric measures, and there was no difference in the use of antibiotics and hospitalizations. PMID- 7478783 TI - The effect of inhaled adrenaline on lung function of recurrently wheezy infants less than 18 months old. AB - Inhaled bronchodilators have been shown not to improve lung function in infants with wheeze. This observation has led to the suggestion that airway wall edema may be more important than bronchoconstriction in infants with airway narrowing. Inhaled adrenaline is used to relieve upper airway edema in children with croup and has been demonstrated to improve clinical scores and lower pulmonary resistance in some infants with wheeze associated with bronchiolitis. The aim of the present study was to examine the effect of inhaled adrenaline on lung function in a group of infants with recurrent wheeze. Eleven infants aged 10 to 18 months with a history of recurrent wheeze were studied during an asymptomatic interval. Respiratory function was assessed (1) by measuring maximal expiratory flow at functional residual capacity (VmaxFRC) during a forced partial expiratory maneuver and (2) by measuring conductance of the respiratory system (Grs) using a single expiratory occlusion technique. Following baseline measurements, the infants received 0.5 mg/kg adrenaline by nebulizer and serial lung function tests were repeated at 5 min intervals. Ten infants had abnormal baseline lung function (median VmaxFRC 44.2% predicted; median Grs 34% predicted). Using a random effects model, VmaxFRC and Grs declined significantly at 10 and 5 min after adrenaline, respectively. No significant improvements from baseline were observed in either measurement for up to 30 min following adrenaline delivery. It is concluded that inhaled adrenaline did not relieve airways obstruction in this group of asymptomatic infants with recurrent wheeze. PMID- 7478784 TI - Two mechanisms for IgG uptake in cultured human trophoblast: evidence for a novel high affinity Fc receptor. AB - The mechanism of IgG transport by the placental trophoblast was examined by studying IgG uptake by purified trophoblast maintained in culture. This model retains the ability to bind and endocytose human IgG from human serum. Comparison of the relative IgG uptake by the trophoblast among the four subclasses of both human and mouse IgG indicates that the trophoblast IgG receptor has different affinities from those described for the three known human Fc gamma receptors, FcR gamma I, FcR gamma II, and FcR gamma III. These results suggest the presence of a novel trophoblast Fc gamma receptor. Although Fc gamma RIII has been reported to be present on trophoblasts, immunocytochemical studies failed to detect binding to the cell surface of antibody-specific for Fc gamma RIII, 3G8 MAb. In addition, blocking studies with MAb 3G8 did not interfere with IgG uptake. Scatchard analysis of human IgG uptake revealed a biphasic curve consistent with two distinct mechanisms for the transport of IgG by the trophoblast. The first is a higher affinity system (Ka = 1.7 x 10(7) M-1, 1.7 x 10(4) binding sites/cell) which exhibits IgG subclass and species specificity, and the second is a low affinity system (Ka = 6.9 x 10(3) M-1, 7.5 x 10(7) binding sites/cell). PMID- 7478785 TI - Prenatal diagnosis of glycogen storage disease type II: enzyme assay or mutation analysis? AB - Two mutations in the lysosomal alpha-glucosidase gene, a single base pair deletion (delta T525) and a deletion of exon 18, have recently been identified with a relatively high incidence in Caucasian patients with glycogen storage disease type II (GSD II). Prenatal diagnosis was made in a pregnancy of consanguineous parents of a child with GSD II. The delta T525 deletion was demonstrated in this family but unexpectedly in only one of the parents. The absence of the delta T525 deletion in DNA isolated from the chorionic villi and a normal alpha-glucosidase activity indicated that the fetus was not affected. The possible role of mutation analysis in the prenatal diagnosis of GSD II is discussed in the light of our previous experience from a series of 100 prenatal diagnoses for this disorder by enzyme analysis. PMID- 7478786 TI - Intrauterine hypoxia-ischemia increases N-methyl-D-aspartate-induced cGMP formation and glutamate accumulation in cultured rat cerebellar granule cells. AB - Effects of intrauterine hypoxia-ischemia (HI) on brain functional development in the term rat were examined in cerebellar granule cell cultures obtained from an in utero HI model. On gestation d 17, HI conditions were achieved by complete clamping of the uterine vasculature for designated durations followed by removal of the clamps to permit reperfusion. Sham operation (surgery without vasculature clamping) was performed as the control. After surgery, the uterine horns were returned to dam's abdomen to let the pups deliver naturally. Severe HI insult from the surgical manipulation was evident in that the lactate levels of fetal brain increased, and fetal blood pH decreased with the duration of vasculature clamping up to 1 h. The experimental HI insult up to 1 h did not affect fetal survival rate, but retarded growth of fetuses or newborns was observed in the 1 h HI group. N-Methyl-D-aspartate (NMDA)- and kainate (KA)-stimulated cGMP formation and glutamate accumulation were measured in cerebellar granule cell cultures from 8-d-old pups suffering from various durations of antenatal HI insult. NMDA (100 microM)-induced elevation of cGMP was further augmented by a 10-35-min HI insult as compared with the control cells (62.4-78.2 versus 49 pmol/mg protein). In the presence of L-NG-monomethyl-arginine (L-NMMA, 150 microM), a nitric oxide synthase inhibitor, NMDA-induced cGMP formation was blocked, and the blockade of cGMP formation by L-NMMA (10 microM) could be reversed by simultaneous application of 1 mM arginine in both control and HI cells.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7478787 TI - Comparative differences and combined effects of interleukin-8, leukotriene B4, and platelet-activating factor on neutrophil chemotaxis of the newborn. AB - IL-8, leukotriene B4 (LTB4), and platelet-activating factor (PAF) are potent neutrophil chemoattractants that have been identified in inflammatory conditions of the newborn such as chronic lung disease of extreme prematurity. The aims of this study were to determine the relative potency and combined effects of these mediators on chemotaxis of polymorphonuclear leukocytes (PMN) from the newborn and to compare the effect of combining all three mediators on chemotaxis of PMN from newborns and adults. Neutrophils were isolated from cord blood (n = 17) or healthy adults (n = 4) and incubated in a 3-tier, 48-well chemotaxis chamber. For PMN from newborns, using chemoattractant concentrations ranging from 0.01 to 100 nM, we found that there were significant differences in potency: IL-8 > LTB4 > PAF. Migration to each of these mediators was almost completely due to chemotaxis as opposed to chemokinesis. At submaximal chemotaxis, using equally effective doses of IL-8 (0.2 nM), LTB4 (1.0 nM), and PAF (10 nM), the combination of all three mediators increased chemotaxis 2.4-fold above the average individual responses. Further studies indicated this increase in chemotaxis was due to the combination of IL-8 and PAF or IL-8 and LTB4; but there was no increase in chemotaxis when PAF and LTB4 were combined. The combination of all three submaximal doses of chemoattractants resulted in PMN chemotaxis that was still 36% of the adult response.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7478788 TI - Magnetic resonance imaging of brain edema in the neonatal rat: a comparison of short and long term hypoxia-ischemia. AB - Diffusion-weighted and transversal relaxation time (T2)-weighted magnetic resonance imaging were used to study the relationship between the duration of hypoxia-ischemia [unilateral common carotid artery (CCA) ligation and exposure to 8% oxygen] and the in vivo visualization of brain edema in 7-d-old rats. After CCA ligation, 35 animals were divided into five groups according to the length of exposure to 8% oxygen: no exposure (n = 9), 15 min (n = 12), 30 min (n = 5), and 1 h (n = 9) exposure; six animals served as controls. Diffusion weighted images were acquired 2 h after the hypoxic-ischemic insult, sequential T2 weighted images were recorded for up to 7 d and the outcome was documented by histologic examination at 21 d. The apparent diffusion coefficient of water in the ipsilateral cortex was significantly decreased in all animals recovering from prolonged hypoxic-ischemic insult (30 min and longer), whereas this was the case in only 40% of animals exposed to 15 min of hypoxia. Moreover, T2 prolongation of brain tissue occurred only in the former group. These results indicate transient and reversible alterations of physiologic water compartmentation for short term hypoxia-ischemia, but irreversible edema formation for long term hypoxia ischemia. They support the hypothesis that the duration of hypoxia-ischemia determines whether a vasogenic edema and infarction follows the initial cytotoxic edema. PMID- 7478789 TI - Hematoma-induced enhanced cerebral vasoconstrictions to leukotriene C4 and endothelin-1 in piglets: role of prostanoids. AB - Cerebral hematoma enhances vasoconstriction induced by topical application of the vasoconstrictor agents endothelin-1 (ET-1) and leukotriene C4 (LTC4). We investigated the influence of dilator prostanoids on vasoconstrictions induced by ET-1 and LTC4 in piglets. Newborn pigs anesthetized with alpha-chloralose were fitted with closed cranial windows 4 d after cortical subarachnoid injections of artificial cerebrospinal fluid (aCSF) (control) or blood (hematoma). The responsiveness of pial arterioles to topical application of the vasoconstrictors ET-1 and LTC4 was examined in the control and hematoma groups before and after treatment with indomethacin (5 mg/kg, i.v.). Vasoconstriction to topical application of LTC4 and ET-1 was enhanced by hematoma compared with the control (28 +/- 2% versus 21 +/- 2% for 10(-8) M LTC4 and by 25 +/- 2% versus 15 +/- 1% for 10(-8) M ET-1, respectively). The lower dose of ET-1 (10(-12) M) dilated pial arterioles in the control group by 6 +/- 2%, hematoma blocked this dilation and it was converted to constriction (10 +/- 1%). Indomethacin treatment enhanced vasoconstriction to LTC4 in the control group to a similar constriction to that observed in the hematoma group. Indomethacin treatment also enhanced vasoconstriction to ET-1 in the control group (25 +/- 1% for 10(-8) M) to similar constrictions to those observed in the hematoma group (25 +/- 2% for 10(-8) M). Dilation to the lower dose of ET-1 was blocked and converted to constriction (17 +/- 2% for 10(-12) M) by indomethacin treatment.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7478790 TI - Disturbance of cerebral oxygenation and hemodynamics related to the opening of the bypass bridge during veno-arterial extracorporeal membrane oxygenation. AB - The objective of this study was to investigate changes of cerebral oxygenation and hemodynamics related to opening of the bypass bridge during veno-arterial extracorporeal membrane oxygenation (ECMO). Ten newborn infants and 12 piglets were studied during opening of the bridge for 10 and 1 s, respectively. Changes in cerebral concentration of oxyhemoglobin (cO2Hb), deoxyhemoglobin (cHHb), (oxidized-reduced) cytochrome aa3 (cCyt.aa3), and blood volume (CBV) were continuously measured by near infrared spectrophotometry. Heart rate, arterial O2 saturation (saO2), and mean arterial blood pressure (MABP) were measured simultaneously. In the piglets, central venous pressure (CVP), intracranial pressure (ICP), and left common carotid artery blood flow (CaBF) were also measured. Opening of the bridge for 10 s in the infants resulted in a significant decrease in MABP, saO2, and cO2Hb, whereas cHHb increased. CBV did not change significantly. In piglets biphasic changes were observed for MABP, CaBF, cO2Hb, and CBV, showing an initial decrease followed by a smaller increase. cHHb and CVP showed reverse biphasic changes. ICP increased but saO2 was unchanged. In all cases heart rate and cCyt.aa3 did not change significantly. Opening of the bridge for 1 s resulted in minor changes in only a few variables. In conclusion, opening of the bridge resulted in a decrease of CBV and cerebral O2 supply due to a decrease of cerebral blood flow, followed by a compensatory increase of cerebral O2 extraction and vasodilatation. The return of oxygenated blood after reclosing resulted in an increase of CBV with overcompensation of cerebral O2 supply. PMID- 7478792 TI - Maturation of the contractile response and its endothelial modulation in newborn porcine intrapulmonary arteries. AB - Pulmonary vascular reactivity is thought to be greater in the newborn than adult lung. To determine the influence of the endothelium on smooth muscle cell contractility, responses of rings of isolated intrapulmonary arteries were studied from pigs at birth aged < 2 h, 2 d, 3 d, and 10 d (n = 4 per age group) and from eight adult animals. At birth, the response to KCl (25 mM) and prostaglandin F2 alpha (PGF2 alpha) (3 microM) but not histamine (0.1 mM) was greater in rings with endothelium (E+) than without (E-). The response to PGF2 alpha decreased between birth and 3 d by which time the contraction was less in E+ rings than E-, but L-nitro monomethyl arginine augmented contraction at all ages. In the immature piglets, the response to phenylephrine was less in E+ rings than E-, an effect which was reversed by L-N omega-nitro-L-arginine methyl ester. The response to all contractile agonists increased between 10 d and adulthood. The concentration of plasma endothelin-1 was determined in all animals by RIA and was higher at birth than at 3 d or later. In summary, 1) at birth, endothelium enhanced contractility, when plasma endothelin was greatest, but released NO in the presence of phenylephrine and PGF2 alpha; 2) contractile response to all agonists was small at birth; and 3) a complex interaction existed between the contractile agonist and the effect of endothelial maturation.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7478791 TI - Whole body arginine metabolism and nitric oxide synthesis in newborns with persistent pulmonary hypertension. AB - Despite the potential relevance of the L-arginine-nitric oxide (NO) pathway in the pathophysiology of pulmonary hypertension, no in vivo studies of the kinetics of arginine and NO have been conducted previously in this population. The terminal guanidino N-atom of L-arginine is the precursor for NO, which is oxidized to the stable inorganic nitrogen oxides, nitrite (NO2-) and nitrate (NO3 ). Thus, synthesized NO is detected in serum or urine as NO2- and NO3-. The purpose of this investigation was to compare studies of whole body arginine metabolism twice in nine patients with persistent pulmonary hypertension of the newborn (PPHN), using a primed constant i.v. infusion of L-[guanidino 15N2,5,5(2)H2]arginine and L-[5,5,5(2)H3]leucine, first during acute pulmonary vasoconstriction and again during convalescence, and thereby to characterize quantitative aspects of whole body arginine kinetics and NO production, as estimated from the rate of transfer of the 15N-guanidino-label of arginine to urinary nitrate (15NO3-). Arginine flux rates were 84.1 +/- 8.6 mumol.kg-1 h-1 (mean +/- SEM) during acute pulmonary hypertension and increased to 125 +/- 13.2 (p < 0.05) during convalescence, whereas leucine fluxes were unchanged (168.5 +/- 15 versus 178.8 +/- 10.2 mumol.kg-1 h-1), and comparable to those reported in healthy newborns. During convalescence total urinary nitrate excreted increased by 66% (p < 0.05), urinary 15NO3- increased from 0.29 +/- 0.07 to 0.74 +/- 0.15 mumol.d-1 (p < 0.05), and the rate of plasma arginine conversion to NO increased from 10.3 +/- 2.2 to 45.6 +/- 13 mumol.d-1 (p < 0.05).(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7478793 TI - Vasopressin effects on lung liquid volume in fetal sheep. AB - The normal switch from placental to pulmonary gas exchange at birth requires rapid removal of liquid from the lungs. Previous reports contend that vasopressin may be important in regulating this process, but this notion derives from studies in which fetal sheep received very large doses of vasopressin that yielded plasma concentrations at least 10 times greater than those that have been measured during normal labor. To study the physiologic effects of vasopressin on lung liquid volume in fetal sheep, we made three sets of experiments. First, we measured plasma vasopressin concentrations [VP] in 15 late-gestation fetal sheep, five of which were at various stages of spontaneous labor. [VP] in these fetuses ranged from < 1 (prelabor) to 31 (during labor) microU/mL; postmortem extravascular lung water (EVLW) ranged from 4.5 to 14.5 g/g dry lung tissue. In a second series of studies, we measured EVLW in five sets of near-term (138 +/- 1 d, term = 147 d) twin fetal sheep that received an 8-h i.v. infusion of either isotonic saline (control twin) or AVP (AVP-treated twin) at a rate of approximately 1 (mU/kg)/min. This dose was chosen to mimic [VP] measured in fetuses that had been studied during labor. [VP] did not change in the control twins, whereas [VP] increased from 1.8 +/- 1.0 to 27.7 +/- 3.5 microU/mL in treated twins. There was a small, statistically significant difference in EVLW between twins that received AVP and untreated twins (11.9 +/- 1.8 versus 14.6 +/- 2.8 g/g dry lung).(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7478794 TI - Evaluation of the pulmonary vasculature and dynamics with intravascular ultrasound imaging in children and infants. AB - The ability to assess the pulmonary vasculature in pulmonary vascular disease by hemodynamic or histologic evaluation is limited. We sought to determine the feasibility of intravascular ultrasound techniques in infants and children and to assess simultaneously the morphology and dynamics of pulmonary arteries. Patients were seen in the Department of Pediatrics, Division of Pediatric Cardiology, Sophia Children's Hospital, Rotterdam, The Netherlands. We performed intravascular ultrasound imaging in 11 pediatric patients with congenital heart disease undergoing cardiac catheterization. Luminal diameter, area, and pulsatility were determined at two to five sites in the pulmonary branches. Pulmonary vascular reaction to 100% oxygen inhalation was studied. Patients weighed 4.1-51.0 kg (21.8 +/- 16.3 kg, mean +/- SD). Luminal diameters, areas, and pulsatilities could be determined reproducibly in arteries with diameters from 1.6 to 9.3 mm. In total 39 sites were studied in 11 patients. Pulsatility was related to vessel size (r = 0.81), although a substantial interindividual variation was present. After 100% oxygen inhalation, pulsatility increased in all arteries (from 20.0 +/- 3.3% to 25.9 +/- 2.9%, p < 0.05), and vasodilatation could be directly visualized, most prominently in the smallest arteries (percentage change in diameter, mean 7.4 +/- 2.8% versus -2.8 +/- 3.1% in the largest arteries, p < 0.001). Measurement of wall thickness was not feasible, but specific changes in the appearance of the wall structure could be recognized in a patient with severely elevated pulmonary vascular resistance. The specific advantages of intravascular ultrasound in assessing pulmonary vascular disease are discussed.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7478795 TI - Alteration of sulfation of glycoconjugates, but not sulfate transport and intracellular inorganic sulfate content in cystic fibrosis airway epithelial cells. AB - The secreted and cell surface high molecular weight glyco-conjugates (HMG) generated by primary cultures of airway epithelial cells from cystic fibrosis (CF) patients are oversulfated. To determine whether this abnormality is maintained in transformed CF airway epithelial cells and whether differences in transport or intracellular accumulation of sulfate can explain this alteration, we assessed sulfate metabolism in paired CF and normal cell lines as well as primary cultures of CF and normal cells. Both 4-acetamido-4'-isothiocyanostilbene 2,2'-disulfonic acid-inhibitable and -resistant [35S]sulfate efflux and influx were identical for each pair of CF and normal cell lines. Furthermore, cell content of inorganic sulfate was not significantly different in CF and normal cells. However, compared with primary CF cells that oversulfate HMG transformed CF cells oversulfated cell surface HMG but not HMG released into culture medium. Our results suggest that plasma membrane sulfate transport is not altered in CF airway epithelial cells and the abnormal sulfation of HMG may be due to perturbation in intracellular sulfate activation or transfer of activated sulfate to HMG. The relationship of this abnormality to CF transmembrane conductance regulator mutations remains to be determined. PMID- 7478796 TI - Secretion of transforming growth factor-alpha (TGF alpha) by postnatal rabbit alveolar macrophages. AB - Transforming growth factor-alpha (TGF alpha) is a cytokine secreted by stimulated alveolar macrophages (AM) in vitro and after in vivo particulate or hyperoxia exposure and has been implicated in the processes of postnatal lung development. It is unknown if AM TGF alpha secretion changes during normal postnatal lung development. After sacrifice of New Zealand white rabbits on postnatal d 0-2, 5 7, 9-10, 14, 21, and 28 and > 4 mo (adult), AM were isolated by discontinuous density centrifugation and placed in culture in the presence or absence of concanavalin A (ConA) for 24 h. Media were collected, and the concentration and isoforms of TGF alpha in AM media samples were determined by an epidermal growth factor/TGF alpha radioreceptor assay and Western immunodetection, respectively. TGF alpha was present in media of AM from the 1.06 and 1.08 g/dL Percoll densities, but not in the 1.10 g/dL density. Statistically significant differences in TGF alpha secretion by unstimulated and ConA-stimulated AM at the various ages were not detected until d 14 (p < 0.02). Western blot analysis of unstimulated AM media samples from d 0-7 rabbits demonstrated the presence of TGF alpha isoforms at 46, 30, and 14.3 kD. At later postnatal ages (> or = d 9), a single 14.3-kD isoform was present. In contrast, analysis of ConA-stimulated AM media samples showed TGF alpha isoforms at 46, 30, and 14.3 kD for all ages; however, the 6-kD mature isoform was present only in juvenile (d 28) and adult media.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7478797 TI - Regulation of sympathetic activity in children with various congenital heart diseases. AB - We studied the alpha- and beta-adrenoceptor activity and catecholamine and cAMP levels in 112 children and infants admitted to the hospital for diagnostic or interventional catheterization of tetralogy of Fallot, ventricular septal defects with or without hypertension, pulmonary stenosis, coarctation of the aorta, and various complex cyanotic congenital cardiac diseases and compared them with 14 children undergoing transcatheter occlusion of patent ductus arteriosus with insignificant left-to-right-shunts. The mean total platelet alpha-adrenoceptor density of the study population was elevated by 73%. Both the increases in acyanotic (p < 0.05) and cyanotic (p < 0.005) patients as well as the difference between the two groups (p < 0.01) were significant. Based on the congenital disease classification, the elevation in receptor density was also significant in all groups of patients, except coarctation of the aorta. On the other hand, the mean lymphocyte beta-adrenoceptor density was attenuated by 27%, showing significant difference between the acyanotic and the patent ductus arteriosus groups, but none between acyanotic and cyanotic or cyanotic and the patent ductus arteriosus groups. Among the congenital groups, only the left-to-right shunts and the pulmonary stenosis group showed significant (p < 0.05) decrease in beta adrenoceptor density, whereas the affinity of all the groups toward [125I]iodocyanopindolol was hardly influenced. The plasma levels of all three catecholamines, norepinephrine, epinephrine, and dopamine, were elevated, but cAMP remained unchanged.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7478798 TI - The role of the upper airway in contributing to respiratory responses during experimental seizures in piglets. AB - Recent data demonstrating stimulation of respiration during seizures are at odds with the frequent clinical finding of respiratory impairment in patients with generalized seizures. To determine the role of the upper airway in contributing to these clinical observations, a study was performed in 12 weanling piglets. An arterial catheter and epidural electrodes were placed in ketamine-anesthetized piglets. In intact piglets, all airflow was measured through a snug-fitting nasal mask whereas pressure was measured with a subglottic catheter. Tracheostomized piglets had airflow and pressure measured at the trachea. Seizures were induced with i.v. injections of pentylenetetrazol. Peak inspiratory flow resistance was calculated by averaging three epochs of five to seven consecutive breaths. Epochs of greater than 20 s in duration with a tidal volume of < 10 mL were termed apnea. Apnea was considered central when airway pressure was > -2 cm H2O and obstructive when < -10 cm H2O. After 20 min of untreated seizures, intact piglets had significant respiratory and metabolic acidosis, whereas tracheostomized piglets had significant increases in Ve and mild metabolic acidosis. Apnea and obstruction were observed frequently in intact piglets. Resistance was unchanged during seizures in tracheostomized piglets, whereas a nearly 4-fold increase in inspiratory resistance was observed in intact piglets. We conclude that upper airway patency and resistances are important determinants of respiratory responses during generalized seizures in piglets. PMID- 7478799 TI - Circulatory and metabolic effects of hypoxia in the hyperinsulinemic ovine fetus. AB - Fetuses of women whose diabetes is poorly controlled often exhibit hypoxemia and elevated catecholamine concentrations at birth. In the ovine fetus, experimental hyperinsulinemia results in hypoxemia, elevated catecholamine concentrations, and hemodynamic changes. Limited oxygen availability occurring during pregnancy related complications and/or delivery may present an additional risk to the hyperinsulinemic fetus. In this study, we tested the hypothesis that hypoxia induced via acutely limiting oxygen availability compromises the hemodynamically and metabolically stressed but compensated hyperinsulinemic ovine fetus. Fetuses receiving insulin (n = 8) or placebo (n = 5) for 48 h were exposed to maternally induced hypoxia. Hypoxic hypoxia was associated with a surge in catecholamines in the hyperinsulinemic fetuses. During hypoxia, this group exhibited insulin related sustained increases in the combined ventricular output and fetal body blood flow, accentuation of the prior insulin-related increase in blood flow to the heart, decreased systemic oxygen delivery, accentuation of the insulin related increased oxygen extraction, reductions in the insulin-related increased systemic oxygen uptake, sustained increases in regional oxygen delivery to the heart and adrenal glands, reductions in the insulin-related increased delivery to the carcass, and decreased oxygen delivery to the kidneys and gastrointestinal tract. We conclude that, in the hyperinsulinemic ovine fetus, hypoxic hypoxia attenuates the hyperinsulinemia-mediated increased systemic oxygen uptake. Regional oxygen transport is preserved to vital regions (brain, heart, and adrenal glands) by increased perfusion and compromised to certain other regions (kidneys and gastrointestinal tract), because the increases in perfusion are insufficient to offset the limited oxygen availability. PMID- 7478800 TI - Studies of the organ distribution in mice of teniposide liposomes designed for treatment of diseases in the mononuclear phagocytic system. AB - Liposomes can be used for the delivery of drugs in cancer chemotherapy. After i.v. injection liposomes are to a large extent taken up by the mononuclear phagocytic system (MPS). When treating diseases in the MPS, such as the histiocytic syndromes, this property is of potential value for drug targeting and may lead to a more efficient therapy with less systemic toxicity. Teniposide (VM 26) is a potent anti-tumor drug. Its lipophilicity makes it suitable for liposomal formulation. Teniposide liposomes were prepared by dissolving egg phosphatidylcholine and dioleoyl phosphatidic acid (19:1 molar ratio) in methylene chloride together with teniposide. After solvent evaporation, the dry lipid film was dispersed in a glucose solution (50 mg/mL), and size calibration was obtained by filtration through polycarbonate filters. The amount of teniposide incorporated was 2.5 mol%. To investigate the organ distribution, teniposide liposomes containing radiolabeled teniposide or phospholipid were given i.v. to mice. By increasing the size of the vesicles, the MPS uptake could be modulated. When vesicles of 200 nm and 1 and 3 microns were injected, the drug levels in the spleen were increased 2.6-, 6.8-, and 21-fold 40 min after injection, compared with levels after injection of the commercial teniposide formulation. It was concluded that organ distribution of teniposide in mice could be modified by administering the drug in liposomal form with the potential of improved treatment of diseases engaging the MPS. PMID- 7478801 TI - 17 alpha-hydroxyprogesterone, 4-androstenedione, and testosterone profiled by routine stable isotope dilution/gas chromatography-mass spectrometry in plasma of children. AB - Using stable isotope dilution/gas chromatography-mass spectrometry (ID/GC-MS), a physicochemical method, we have profiled the plasma steroids 17 alpha hydroxyprogesterone, 4-androstenedione, and testosterone in normal children of various age groups. Comparison of our values with those obtained by direct immunologic assays and those using an extraction or purification step showed that immunoassays in general overestimate steroid concentrations. This was especially true for plasma samples in the neonatal period and was most expressed for the concentrations of 17 alpha-hydroxyprogesterone. Our study demonstrated the applicability of ID/GC-MS to routine clinical steroid analysis. The application of ID/GC-MS is recommended whenever problems from matrix effects or cross reactivity are likely to arise or suspicious results by immunoassays need to be rechecked. PMID- 7478802 TI - Ontogeny of L-alanine uptake in plasma membrane vesicles from rat liver. AB - Alanine uptake into liver plasma membrane vesicles was studied at different stages of postnatal rat development. Before weaning, alanine hepatic uptake showed lower values for the global KM than after weaning (0.34, 0.77, 1.45, and 1.61 mM for 1-, 15-, and 28-d-old and adult rats, respectively). Alanine uptake capacity increased progressively until reaching maximum values in the adult state (values for Vmax: 0.078, 0.199, 0.317, and 0.613 nmol alanine/mg protein/3 s for 1-, 15-, and 28-d-old and adult rats, respectively). These results seem to point to a prevalence of a high affinity, low capacity alanine transport component (traditionally assumed to be attributable to system A) in newborn and suckling rats, in agreement with our previous results on isolated hepatocytes (Martinez Mas JV, Casado J, Felipe A, Marin JJG, Pastor-Anglada M: Biochem J 293: 819-824, 1993). The suckling-weaning developmental transition seems to play a role in establishing the pattern of adult hepatic alanine transport characterized by a higher capacity but a lower affinity (because most alanine is taken up by system ASC) inasmuch as KM values show a 100% increase after weaning, although Vmax values continue to increase steadily until the adult age. PMID- 7478803 TI - Obesity after successful treatment of acute lymphoblastic leukemia in childhood. AB - Early and late effects of treatment for acute lymphoblastic leukemia (ALL) on weight was retrospectively investigated in 113 children in continuous first remission. Weight was examined at diagnosis up to 10 y after cessation of treatment. There was an increased prevalence of overweight after treatment for ALL which persisted over time. All treatment regimens included corticosteroid therapy, and 52 patients received additional cranial irradiation. Patients treated with and without cranial irradiation did not differ in weight gain, indicating that not cranial irradiation but corticosteroid therapy might explain weight gain in children treated for ALL. Dexamethasone was associated with a significant increase of weight at cessation of treatment. Patients treated with a combination of prednisone and dexamethasone had as a late effect the highest prevalence of obesity (44%). Gender or age at diagnosis were not related to weight gain. PMID- 7478804 TI - Changes in dopaminergic control of circulating melanocyte-stimulating hormone related peptides at puberty. AB - Desacetyl alpha-melanocyte-stimulating hormone (MSH) (ACTH 1-13) is the main form of immunoreactive alpha-MSH circulating in human plasma. This study evaluates the possibility that a dopaminergic inhibitory mechanism could be operative during human development. Thus, alpha-MSH and ACTH 1-13 plasma levels were measured after dopaminergic blockade (domperidone (0.3 mg/kg body weight, maximum 10 mg, p.o.) in 13 prepubertal (aged 4.5-12.3 y) and 12 pubertal (aged 10.2-16.9 y) children. Both peptides were measured by RIA after plasma extraction on Sep-pak C 18 cartridges and reverse phase HPLC. The chromatographic profile of alpha-MSH immunoreactivity falls into two main peaks, corresponding to the retention time of alpha-MSH and ACTH 1-13. Moreover, in prepubertal children domperidone induced a significant increase of alpha-MSH from 1.7 (median) to 5.0 pmol/L, whereas no changes in alpha-MSH plasma levels were found in pubertal subjects (from 5.0 to 4.1 pmol/L). Similarly, ACTH 1-13 plasma levels significantly increased from 3.0 to 19.8 pmol/L in prepubertal children remaining stable in pubertal ones (from 7.8 to 4.6 pmol/L). Moreover, a significant negative correlation was found between basal DHEA-S levels and the plasma alpha-MSH increase after domperidone. These data demonstrate that: 1) ACTH 1-13 is the main form of immunoreactive alpha-MSH in prepubertal life and 2) the dopaminergic inhibition of both ACTH 1 13 and alpha-MSH plasma levels is apparent only in prepubertal subjects. PMID- 7478805 TI - Sickle cell vaso-occlusive crisis is associated with abnormalities in the ratio of vasoconstrictor to vasodilator prostanoids. AB - Plasma levels of 6-keto-prostaglandin F1 alpha (6kPGF1 alpha) and thromboxane (Tx) B2 have been assessed in sickle cell disease (SCD) with discrepant results. Inasmuch as direct measurement of plasma prostanoids is fraught with the problem of interfering substances, we assessed plasma 6kPGF1 alpha and TxB2 levels in patients with SCD by RIA after extraction of eicosanoids and separation by HPLC. We demonstrate that the 6kPGF1 alpha and TxB2 levels in children with SCD in steady state as well as in vaso-occlusive crisis (VOC) are significantly lower when compared with those from age-matched controls. The VOC plasma 6kPGF1 alpha and TxB2 levels were, however, significantly elevated when compared with those from children in steady state. Changes similar to those noted with unpaired plasma samples were also observed when paired steady state and VOC plasmas from the same patients were assessed. The ratio of TxB2 to 6kPGF1 alpha was, however, significantly elevated in patients with SCD in crisis when compared with eicosanoid ratios obtained during steady state. In an attempt to understand whether the abnormality in 6kPGF1 alpha was due to an impairment in endothelial cell prostacyclin-regenerating ability, we compared the ability of plasma from controls and children with SCD to activate arachidonic acid (AA) release and prostacyclin production by [14C]AA-prelabeled bovine aortic endothelial cells. Our results suggest that the decreased 6kPGF1 alpha levels in plasma from children with SCD was not due to an effect on substrate AA release but rather a modulatory effect of sickle plasma components on endothelial cell cyclooxygenase activity.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7478806 TI - Postnatal change in inhibitory regulation of intestinal motor activity in human and canine neonates. AB - Motor activity was recorded in 19 preterm infants three times during the first postnatal month. There was a paucity of small intestinal motor quiescence during fasting in the first postnatal week; however, its duration significantly increased with postnatal age (p < 0.03). Although fasting motor patterns changed with postnatal age, motor responses to feeding were present within the first few days of life. Motor activity was also recorded weekly in 11 newborn dogs for 6 wk. Intestinal motor quiescence was also absent during fasting for the first postnatal week but it significantly increased with postnatal age (p < 0.002). As in the human preterm infant, a motor response to feeding was present within the first few days of life. Plasma concentrations of gastrin and peptide YY during fasting were low in the preterm human and canine neonate during the first postnatal week but plasma concentrations of both peptides increased with postnatal age. Although plasma concentrations of gastrin were low during fasting for the first postnatal week, plasma concentrations of gastrin increased significantly postprandially compared with fasting (p < 0.05). We conclude that motor quiescence during fasting becomes a more prominent feature of newborn intestinal motor function postnatally. In addition the release of two peptides that regulate motor patterns also change postnatally. Thus, postnatal changes in motor patterns and peptide release change in a parallel fashion in human preterm neonates and canine neonates.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7478807 TI - Age-related susceptibility to chlordecone-potentiated carbon tetrachloride hepatotoxicity and lethality is due to hepatic quiescence. AB - Previous studies revealed that postnatally developing rats are resilient to the lethal effects of chlordecone (CD) + carbon tetrachloride (CCl4) combination. The objective of this study was to investigate the underlying mechanism. We hypothesized that ongoing cell division and cell cycle progression as well as additional toxicant-induced stimulation of tissue repair help in restraining the progression of injury on the one hand, and in recovery through speedy healing on the other. Postnatally developing (20- and 45-d) and adult (60-d) male Sprague Dawley rats were challenged with a nontoxic single dose of CCl4 (100 microL/kg, i.p.) or corn oil after pretreatment with either dietary CD (10 ppm) or normal diet (ND) for 15 d. Hepatocellular injury was assessed by measuring serum enzymes [alanine transaminase (ALT), sorbitol dehydrogenase (SDH)], and bilirubin, as well as by histopathologic examination of liver sections during a time course of 0-96 h after the administration of CCl4 or corn oil. Hepatocellular regeneration was assessed by [3H]thymidine ([3H]T) incorporation into hepatic nuclear DNA. In CD+CCl4 treatment, ALT, SDH, and bilirubin levels peaked between 36 and 48 h after CCl4. All 20-d-old rats survived the challenge of CD+CCl4. CD-potentiated hepatotoxicity and lethality of CCl4 begin to be manifested in 45-d-old rats at 48 h and later times (25% mortality), whereas adult rats experience progressive hepatotoxic injury and 100% mortality by 72 h. In contrast, regardless of pretreatment, 20-d-old rats recover fully from injury by 72 h after CCl4 treatment.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7478808 TI - Serum insulin-like growth factor (IGF) binding protein-3 and IGF-I levels during childhood and adolescence. A cross-sectional study. AB - To investigate the effect of pubertal development on serum levels of IGF binding protein-3 (IGFBP-3) and IGF-I, and the relationship between IGFBP-3 levels and height, weight, weight for height and age (WFHA), and IGF-I levels, a cross sectional study was performed in a Spanish basic education school in Vigo (NW Spain). The study was made up of 181 girls with a mean chronologic age of 11.03 +/- 0.22 y and 173 boys with a mean chronologic age of 10.9 +/- 0.23 y. The pubertal development was graded into three groups according to estradiol and testosterone concentrations for girls and boys, respectively. All subjects were in good health and among the 5th and 95th percentile for height. Serum IGFBP-3 and plasma IGF-I concentration was determined by RIA. Pubertal development was significantly associated with IGFBP-3 and IGF-I concentrations in girls and boys, respectively (p < 0.0001, analysis of variance). Multivariate regression analyses between IGF-I or IGFBP-3 with age, sex, and estradiol or testosterone show significative correlation in prepubertal children for IGF-I (r = 0.545, p = 0.0001 and r = 0.574, p = 0.0001 for girls and boys, respectively) and only in prepubertal boys for IGFBP-3 (r = 0.336, p = 0.0012). The linear correlation between IGF-I and IGFBP-3 was significant in both prepubertal (r = 0.25, p < 0.0001) and pubertal (r = 0.40, p < 0.0001) girls, but only in prepubertal boys (r = 0.30, p < 0.0001).(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7478809 TI - Relationship between bile acid transplacental gradients and transport across the fetal-facing plasma membrane of the human trophoblast. AB - Bile acids and bilirubin are synthesized by the fetal liver very early on during intrauterine life. The main fate of these compounds is to be transferred to the mother. This excretory role of the placenta is primarily determined by the ability of the trophoblast to transport them, which is believed to occur mainly by carrier-mediated processes. The aim of this study was to investigate the role of the cholephilic organic anion exchanger located in the fetal-facing plasma membrane of the human trophoblast in placental "biliary-like" function. No relationship between the magnitude of transplacental gradients for total bile acids and bilirubin was found. However, transport studies, which were carried out by using purified plasma membrane vesicles derived from the fetal-facing pole of the human trophoblast, revealed that [14C]taurocholate transport was affected by both another bile acid (taurochenodeoxycholic acid) and a non-bile acid cholephilic organic anion (bromosulfophthalein). On plotting the ability of different major bile acid species to inhibit radiolabeled taurocholate uptake by these vesicles versus their concentrations in fetal serum or the magnitude of their transplacental gradients, inverse relationships were found. Lower fetal serum concentrations and transplacental gradients were found for bile acid species with higher abilities to affect this transport and presumably to interact with the carrier. By contrast, the magnitude of the transplacental gradient for bile acid species was not correlated with their hydrophobic/hydrophilic balance, as would be expected if diffusion across the lipidic structures of the placental barrier would be the major pathway for the flux of bile acid across this organ.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7478810 TI - Differential effects of glucocorticoids and mineralocorticoids on the mRNA expression of colon ion transporters in infant rats. AB - Several epithelial ion transporters are developmentally regulated in the preweaning period, at the time when the circulating levels of glucocorticoid and mineralocorticoid hormones increase. The specific role of glucocorticoids and mineralocorticoids in the maturation of epithelial ion transport is still disputed. In this study, we investigated the effect of corticosteroids on the mRNA expression of ion transporters in the infant rat colon, a glucocorticoid- and mineralocorticoid-sensitive organ. The expression of the Na,K-ATPase, the H,K ATPase and the amiloride-sensitive Na+ channel mRNA was investigated in control rats from fetal to adult life. We found that the mRNA of the three transporters is temporarily up-regulated in the preweaning period. Rats were then injected with a single dose of betamethasone or aldosterone at 10 d of age. The main effect was the glucocorticoid stimulation of the Na,K-ATPase mRNA within 6 h (4 fold). Glucocorticoids did not alter H,K-ATPase nor Na+ channel mRNA within 6 h. Aldosterone moderately (1.7-fold) stimulated Na+ channel within 6 h, but dit not alter Na,K-ATPase nor H,K-ATPase mRNA. Twenty-four hours after injection, both glucocorticoids and mineralocorticoids had less pronounced and distinct effects. In tissue with lower aldosterone receptor abundance (renal cortex) or with no aldosterone receptor (stomach), glucocorticoids induce a similarly rapid increases in Na,K-ATPase mRNA (4-fold within 6 h), whereas aldosterone had no effect within 6 h. However, glucocorticoids did not stimulate Na,K-ATPase mRNA in the brain, a tissue rich in glucocorticoid receptors.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7478811 TI - Effect of hyperinsulinemia on the development of blood coagulation in the lamb fetus. AB - Infants of diabetic mothers have an increased risk for thrombosis. The etiology of their hypercoagulable state is unknown. To examine the effects of hyperinsulinemia on the development of coagulation during fetal life, 10 sets of chronically catheterized fetal lambs were studied. One twin from each pair of 120 d gestation lamb fetuses was infused with insulin at a rate of 2.5 U/h for 48 h, whereas its twin sibling was infused with an equal volume (20 mL) of dextrose 5% in water. Changes in coagulation factor activities were measured before and after the infusions, and differences were analyzed by paired t tests. There was a significant decrease in protein C after insulin treatment in the insulin-treated twins. There were relative increases in fibrinogen factors V, VII, and XI when the insulin-treated group was compared with the controls. The changes are consistent with an increased risk of thrombosis and may explain, in part, the higher incidence of thrombosis in infants of diabetic mothers. PMID- 7478812 TI - Oral transforming growth factor-alpha enhances jejunal mucosal recovery and electrical resistance in piglet rotavirus enteritis. AB - A randomized, investigator-masked trial determined the effects of oral recombinant human transforming growth factor-alpha (TGF alpha) on jejunal mucosal recovery in 75 piglets with rotavirus diarrhea. Rotavirus inoculation of artificially reared piglets induced subtotal (approximately 50%) villus atrophy and watery diarrhea. Dietary TGF alpha was associated with significant restoration of villus surface area by 4 d postinoculation (p.i.) and complete restoration by 8 d p.i., whereas saline-treated animals required 12 d for recovery. Jejunal segments from clinically recovered TGF alpha-treated piglets showed an increase in electrical resistance across the epithelial barrier in vitro which was proportional to villus height. TGF alpha treatment for 12 d also produced a 30-50% increase in jejunal mucosal mass (protein content and wet weight), compared with the corresponding values in saline-treated piglets and in uninfected controls. However, oral TGF alpha did not hasten the resolution of diarrhea, enhance the specific activities of jejunal mucosal digestive enzymes, or increase jejunal glucose-stimulated Na+ absorption in vitro. We conclude that dietary TGF alpha stimulates jejunal mucosal hypertrophy, improves barrier function, and enhances regrowth of villi in rotavirus enteritis; however, it does not facilitate the restoration of functional activity or mucosal digestive enzymes. Oral TGF alpha can facilitate intestinal epithelial recovery in diseases associated with mucosal damage. PMID- 7478813 TI - Increased proportions of bacteria capable of cleaving IgA1 in the pharynx of infants with atopic disease. AB - Based on the observation that children with a history of atopic disease show significantly increased levels of cleaved secretory IgA in nasopharyngeal secretions, we have previously formulated the hypothesis that bacteria-induced local deficiencies of the immune barrier of the upper respiratory tract may be a contributing factor in the development and perpetuation of atopic diseases. To evaluate this hypothesis, 25 infants were subjected to clinical, bacteriologic, and immunologic examination at the age of 18 mo, 30 mo, and 5 y. The 11 infants, who showed clinical and immunologic evidence of atopic disease at the age of 18 mo, harbored significantly higher proportions of IgA1 protease-producing bacteria (median, 36%; range, 14-64%) than the 14 healthy infants (median, 5%; range, 0.4 14%). No statistically significant differences were observed at the two subsequent examinations, but healthy children showed a statistically significant increase in proportions of IgA1 protease-producing bacteria in the pharynx with increasing age. IgA1 protease-producing bacteria detected included Streptococcus mitis biovar 1, Haemophilus influenzae, Haemophilus parahaemolyticus, Streptococcus pneumoniae, and Neisseria meningitidis, of which the first mentioned species was mainly responsible for the difference observed at the 18-mo examination. Percentage proportions of IgA1 protease-producing bacteria were significantly related to passive smoking which may stimulate the premature and more pronounced pharyngeal colonization of the atopic infants with the IgA1 protease-producing variant of S. mitis biovar 1. The results of the study support the hypothesis that IgA1 protease-producing bacteria colonizing the upper respiratory tract jeopardize the local immune barrier and, thereby, may facilitate the penetration of potential allergens resulting in atopic disease. PMID- 7478814 TI - Pattern of cytokine production by T cells from adolescents with asthma in remission, after stimulation with Dermatophagoides farinae antigen. AB - Children with asthma usually become asymptomatic by the time they reach age 20 y. To clarify the immunologic mechanisms responsible for this phenomenon, we studied patients in remission and others who still had frequent asthma attacks. Patients were grouped by clinical status, and three variables were measured: serum levels of IgE, production of IL4 and interferon (IFN)-gamma, and the activation of T cells induced by Dermatophagoides farinae (Df) antigen. Df-induced activation of T cells (as measured by antigen-induced IL2 responsiveness) or IL2 synthesis itself was induced in patients with active asthma but not in normal subjects. These responses were much weaker in patients in remission. When stimulated by Df antigen in vitro, lymphocytes from patients with active asthma produced much more IL4 than did the cells from normal subjects, and cells from patients in remission produced only a small amount. In contrast, under similar conditions lymphocytes from patients with active asthma produced less IFN-gamma than did the cells from normal subjects. Production of IFN-gamma stimulated by Df antigen was high in patients in remission but not in normal subjects. Thus, upregulated IFN-gamma production after exposure to Df antigen might reduce IL4 secretion, which would suppress IgE production and would improve clinical status. Df antigen may suppress Df-induced allergic responses in patients with asthma in remission. PMID- 7478815 TI - Pain in children with juvenile rheumatoid arthritis: a descriptive study. AB - The purpose of this study is to describe the pain experienced by children with juvenile rheumatoid arthritis. A patient sample composed of 33 children, 7-16 y of age, with juvenile rheumatoid arthritis was included in the study. The children, their parents, and the attending rheumatologist assessed the child's pain using the Abu-Saad pediatric pain assessment tool. In addition, a disease activity index was used by the physician. All children reported pain. This was most often described as hurting, stinging, warm, and uncomfortable. Significant correlations were found for present pain between the child, parent, and physician. The worst pain of the child for the previous week, and not the present pain, correlated with the disease activity as rated by the physician. These findings indicate a need for pain assessment in patients with juvenile rheumatoid arthritis and emphasize the importance of longitudinal studies in this area. PMID- 7478817 TI - Beta-endorphin may be a mediator of apnea induced by the laryngeal chemoreflex in piglets. AB - To determine whether beta-endorphin is involved in the laryngeal chemoreflex, we initially injected 0.01-1 mg of beta-endorphin into the cisterna magna (i.c.m.) and registered the respiratory and cardiovascular patterns in 5-10-d-old piglets. From 0.1 to 1 mg of beta-endorphin i.c.m. induced a decrease in the minute volume, heart rate, and blood pressure within 15 min. Within the next 30 min respiratory pauses accompanied by blood pressure increases and reductions in heart rate developed, similar to the respiratory and cardiovascular pattern of the induced laryngeal chemoreflex. Based on these initial data, we decided to induce a laryngeal chemoreflex in piglets pretreated with 0.1 mg of beta endorphin i.c.m (n = 6), 0.2 mg of beta-endorphin i.c.m. (n = 6), 0.1 mg of beta endorphin i.c.m. and 100 micrograms/kg naloxone i.v. (n = 6), 100 micrograms/kg naloxone i.v. (n = 6), or water i.c.m. (n = 6). Because elevated levels of hypoxanthine in the vitreous humor may indicate hypoxia before death, we therefore measured the postmortem hypoxanthine levels in the vitreous humor. The laryngeal chemoreflex-induced apnea was shortened in the piglet group pretreated with water i.c.m and naloxone i.v. (p < 0.01) and in the piglet group pretreated with 0.1 mg of beta-endorphin i.c.m and naloxone i.v. (p < 0.05), but not significantly prolonged in the piglet groups pretreated with 0.1 or 0.2 mg of beta-endorphin i.c.m. when compared with the piglets pretreated with water i.c.m.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7478816 TI - Inhaled nitric oxide versus inhaled prostacyclin and intravenous versus inhaled prostacyclin in acute respiratory failure with pulmonary hypertension in piglets. AB - This study was a prospective, randomized design to compare oxygenation and pulmonary hemodynamics between inhaled nitric oxide (NO) and inhaled prostacyclin (PGI2), and between inhaled and i.v. PGI2 in acute respiratory failure with pulmonary hypertension. Acute respiratory failure with pulmonary hypertension was induced in 12 piglets weighing 9-12 kg by repeated lung lavages and a continuous infusion of the stable endoperoxane analogue of thromboxane. Thereafter the animals were randomly assigned either for NO or PGI2 application. All animals were treated with different concentrations of NO or different doses of PGI2 applied i.v. and inhaled in random order. Continuous monitoring included ECG, central venous pressure (CVP), mean pulmonary artery pressure (MPAP), mean arterial pressure (MAP), arterial oxygen saturation (SaO2), and mixed venous oxygen saturation (SvO2) measurements. NO inhalation of 10 ppm resulted in a significant increase in PaO2/fraction of inspired oxygen (FiO2) from 7.8 +/- 1.34 kPa to 46.1 +/- 9.7 kPa. MPAP decreased significantly from 5.1 +/- 0.26 kPa to 3.7 +/- 0.26 kPa during inhaled NO of 40 ppm; i.v. infusion of PGI2 slightly increased oxygenation parameters. A significant increase in PaO2/FiO2 up to 32.4 +/- 3.1 kPa was observed during PGI2 aerosol delivery (p < 0.01); i.v. PGI2 decreased MAP from 11.5 +/- 0.39 kPa to 9.8 +/- 0.66 kPa (p < 0.05) and MPAP from 5.8 +/- 0.53 kPa to 4.5 +/- 0.66 kPa, respectively (p < 0.05). PGI2 aerosol delivery significantly decreased the MPAP to 3.7 +/- 0.53 kPa (p < 0.05) without influencing the MAP.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7478818 TI - Positive end-expiratory pressure during KL4 surfactant instillation enhances intrapulmonary distribution in a simian model of respiratory distress syndrome. AB - Intrapulmonary distribution of a peptide-phospholipid (KL4) surfactant administered through an adapter permitting maintenance of positive end-expiratory pressure was compared with distribution by instillation with disconnection from mechanical ventilation in 10 surfactant-deficient Macaca mullata preterm infants. Animals received KL4 surfactant (200 mg/kg) when the arterial to alveolar (oxygen ratio) (a/Ao2) was < or = 0.22 (approximately 50 min after birth) on mechanical ventilation. Six rhesus infants received bolus instillation of two half doses of KL4 surfactant through an endotracheal tube adapter over 10-15 s while maintaining positive end-expiratory pressure (0.4 kPa) accompanied by turning to the right and left lateral positions for 60 s. In four rhesus premature infants KL4 surfactant was injected as two half-dose boluses through the endotracheal tube with disconnection from mechanical ventilation while positioning the infant rhesus monkey in the right and left lateral positions for 30 s of mechanical ventilation between instillation. Acute effects on oxygen saturation were monitored, and physiologic measures of a/Ao2, mean airway pressure, and the ventilatory efficiency index were monitored over the 12-h study. Intrapulmonary distribution of KL4 surfactant was determined using dye-labeled microspheres or [3H]dipalmitoylphosphatidylcholine-labeled surfactant, measured by colorimetry or by scintillation counting. Lungs of each monkey were processed into 50 +/- 5 pieces to determine distribution of radiolabel or microspheres and for scanning electron microscopy. The drop in oxygen saturation was greater among monkey infants disconnected from the ventilator for surfactant instillation.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7478819 TI - Assessment of tidal breathing patterns for monitoring of bronchial obstruction in infants. AB - Two parameters of tidal breathing, the ratio of time to reach peak tidal expiratory flow to the total expiratory time (Tme/TE) and the ratio of volume exhaled at peak tidal expiratory flow to the total exhaled volume (dV/VT) were used to assess lung function in 21 sedated infants (aged 6-14 mo) with different degrees of airway obstruction. These parameters were compared with airway resistance as percentage predicted (Raw%) and maximum expiratory flow at functional residual capacity corrected for lung volume (VmaxFRC/TGV). VmaxFRC/TGV values correlated significantly with Tme/TE (r = 0.630, p = 0.002) as well as with dV/VT (r = 0.728, p = 0.001). Raw% values showed only a weak correlation with dV/VT (r = -0.435, p = 0.048). We conclude that Tme/Te and dV/VT are both able to detect airway obstruction in infants and that these parameters correlate much better with the forced expiratory flow values obtained by the rapid thoracic compression method than with airway resistance, determined by body plethysmography. PMID- 7478820 TI - Reducing equivalent shuttles in developing porcine myocardium: enhanced capacity in the newborn heart. AB - The metabolic demands of the newborn heart are met primarily by glucose and lactate. Mitochondria are impermeable to the NADH produced by these cytosolic reactions. The malate/aspartate and alpha-glycerophosphate (alpha-GP) shuttles provide two pathways to transport reducing equivalents into mitochondria. The goals of this study were to compare the capacity of these shuttles in newborn and adult cardiac mitochondria and to measure the maximal activity of the mitochondrial enzymes involved in these shuttles. Shuttle and enzyme capacities were measured in isolated mitochondria from the left and right ventricular free wall of 0-3-d-old and adult pig hearts. Malate/aspartate shuttle capacity was nearly three times greater in the newborn left ventricle compared with adult (newborn, 616 +/- 24; adult, 232 +/- 28 nmol/min/mg; mean +/- SEM; n = 8; p < 0.00001). The capacity of the malate/aspartate shuttle of the right ventricular free wall was greater than the left in the adult heart. Despite a decrease in malate/aspartate shuttle capacity, maximal activity of mitochondrial matrix enzymes involved in this pathway were increased in adult mitochondria. alpha-GP shuttle activity was absent in adult myocardium. Newborn left ventricular myocardium had significant alpha-GP shuttle activity (44 +/- 4 nmol/min/mg) due to enhanced flavin-linked mitochondrial alpha-GP dehydrogenase activity compared with adult. Interventricular differences in the alpha-GP shuttle capacity were not found in newborn or adult hearts. These findings suggest a mechanism for the substrate preference of neonatal myocardium. PMID- 7478821 TI - Mechanical and metabolic characterization of ischemic contracture in the neonatal pig heart. AB - Isolated, paced, isovolumetrically beating piglet hearts (n = 37) underwent retrograde aortic perfusion with a crystalloid solution during three periods: 1) baseline (coronary perfusion pressure 60 mm Hg), 2) ischemia (coronary flow 10% of baseline for approximately 80 min), and 3) reperfusion (perfusion pressure returned to baseline). In one group of hearts, glycolysis (using 3H2O formation from [3H]glucose) was assessed. During baseline, peak systolic pressure (PSP) was 101.1 +/- 5.0 mm Hg, end diastolic pressure (EDP) 4.4 +/- 0.5 mm Hg, glycolysis 970.5 +/- 65.3 nmol/min/gwet, and myocardial glycogen 234.8 +/- 12.0 mumol/gdry. During ischemia, PSP decreased to 23.3 +/- 2.7 mm Hg, EDP increased to 12.3 +/- 0.7 mm Hg, myocardial glycogen decreased to 181.5 +/- 30.3 mumol/gdry, and lactate (approximately 154 mumol/gwet) and glycerol (approximately 930 nmol/gwet) were released. Myocardial contracture correlated with a decrease in lactate release. Glycolysis decreased to approximately 400 nmol/min/gwet and remained stable, accounting for approximately 50% of the lactate produced. During reperfusion, PSP recovered to 79.8 +/- 3.5 mm Hg, EDP 6.6 +/- 1.7 mm Hg, and glycolysis 1103.9 +/- 81 nmol/min/gwet. In a second group of hearts, with similar mechanical responses, glucose oxidation (using 14CO2 formation from [14C]glucose) was evaluated. During baseline, glucose oxidation was 165.4 +/- 15.9 nmol/min/gwet and correlated closely (r = 0.957) with mechanical activity. With ischemia, glucose oxidation decreased to approximately 17 nmol/min/gwet, yet accounted for approximately 42% of the ATP produced. Upon reperfusion, glucose oxidation returned to baseline values, but now correlated poorly (r = 0.574) with mechanical activity.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7478823 TI - Effect of indomethacin on cerebral blood flow and oxygenation in the normal and ventilated fetal lamb. AB - Indomethacin lowers fetal and neonatal brain blood flow and may reduce the risk of periventricular-intraventricular hemorrhage. However, concerns have been raised that cerebral O2 metabolism may be compromised at lower cerebral perfusion pressures. In 17 near-term lamb fetuses, changes in brain blood flow and cerebral O2 metabolism (CMRO2) were measured at mean carotid arterial pressures (MCBP) ranging from 8 to 70 mm Hg. MCBP was adjusted by inflating balloon occluders around the aortic isthmus and brachiocephalic trunk. This was done before and during intrauterine pulmonary ventilation and oxygenation. Nine fetuses were pretreated with indomethacin (1 mg/kg i.v.); eight served as control. Changes in brain blood flow were assessed from carotid arterial blood flow (Qcar, mL/min) measured with flow transducers. In 15 animals, brain blood flow was also measured intermittently by radionuclide-labeled microspheres (Qbrain). Qcar correlated closely with Qbrain (r = 0.94, p < 0.0001); this relationship was not altered by indomethacin or by ventilation with oxygen. In the nonventilated fetuses, indomethacin decreased Qcar at pressures above the lower limit of cerebral autoregulation (43 mm Hg). However, at MCBP below 44 mm Hg, Qcar with indomethacin was not significantly different from controls. CMRo2 fell when MCBP was decreased below 30 mm Hg (range 8-29 mm Hg), but there was no significant difference between control and indomethacin-pretreated fetuses. In the ventilated fetuses, indomethacin reduced the slope of the pressure-flow relationship above the lower limit of cerebral autoregulation (43 mm Hg), suggesting improved cerebral autoregulation. When MCBP was decreased below 44 mm Hg (range 10-43 mm Hg), indomethacin did not lower Qcar or CMRO2 as compared with controls.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7478822 TI - Nitric oxide and beta-adrenergic mechanisms modify contractile responses to norepinephrine in ovine fetal and newborn cerebral arteries. AB - Ovine fetal cerebral arteries exhibit an enhanced contractile response to norepinephrine (NE) compared with newborns and adults. It is possible that beta adrenergic receptors and/or nitric oxide (NO), a putative endothelium-dependent relaxing factor, differentially modulate cerebrovascular responsiveness to NE as a function of development. The present study evaluated the effect of the beta adrenoceptor antagonist, propranolol, and the NO synthase inhibitor, NG-nitro-L arginine methyl ester (LNAME), on the contractile response of isolated middle cerebral artery (MCA) and basilar artery (BA) to NE during fetal development. MCAs isolated from four preterm fetal lambs (105 d of gestation), seven near-term fetal lambs (125-130 d of gestation), and eight newborn lambs (2-7 d of age) were evaluated using organ baths. BAs isolated from the near-term fetal and newborn lambs were also evaluated. Contractile reactivity of MCAs to NE decreased significantly during fetal maturation as manifested by a marked decrease in Fmax (maximal relative contractile force generated) and an increase in EC50 (Fmax = 100 +/- 7, 41 +/- 7, and 28 +/- 8% of KCl contraction; EC50 = 0.14 +/- 0.03, 1.09 +/- 0.36, and 1.07 +/- 0.22 microM for preterm fetus, near-term fetus, and newborn lamb MCAs, respectively, p < or = 0.05). Propranolol treatment (10(-5) M) increased Fmax (2-fold) only for newborn lamb MCAs. Pretreatment with LNAME (10( 4) M) markedly enhanced the contractile response to NE (7-fold decrease in EC50 and 2-fold increase in Fmax, p < 0.05) for near-term fetus MCAs, whereas preterm fetus and newborn lamb MCAs were unaffected by the inhibitor.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7478824 TI - Effect of status epilepticus on hypoxic-ischemic brain damage in the immature rat. AB - Seven-day postnatal rats were subjected to unilateral common carotid artery ligation, 3 h after which they were subjected to hypoxia with 8% oxygen at 37 degrees C for 2 h. Thereafter, they received multiple s.c. injection(s) of bicuculline (6 mg/kg) adequate to produce behaviorally apparent seizures lasting greater than 1 h (status epilepticus). Repeated episodes of status epilepticus at 2, 6, and 12 h of recovery from hypoxia-ischemia (HI) produced a mortality rate of 53%. Among the survivors, there was no statistically significant difference in the extent of brain damage between convulsing and non-convulsing HI controls, analyzed at 30 d of age. Histopathologic examination for acute lesions also indicated no difference in the severity of brain damage between dead and surviving rat pups subjected to status epilepticus, indicating that mortality was not related to the severity of prior HI brain damage. Those immature rats that died during status epilepticus exhibited lower blood glucose concentrations (1.75 +/- 0.35 mmol/L) compared with surviving, convulsing animals (4.25 +/- 0.51 mmol/L; p = 0.016). Glucose supplementation (0.1 mL of 50% glucose) early during status epilepticus improved survival and significantly prolonged seizure activity (90 +/- 14 min) compared with non-glucose treated, convulsing littermates (47 +/- 10 min; p = 0.02). Glucose supplementation did not increase the extent of brain damage despite improved survival and increased duration of seizure activity.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7478825 TI - Visual evoked potential abnormalities in jaundiced Gunn rats treated with sulfadimethoxine. AB - The manifestations of bilirubin encephalopathy include disturbances in the visual pathway (visual gaze paralysis and distorted visual perception). In the young jaundiced Gunn rat (jj) model of hyperbilirubinemia, significant differences in visual evoked potential (VEP) patterns have been recorded during development. In the present study, the effects of sulfadimethoxine (SDM) on VEP and electroretinogram (ERG) were examined in 3-wk-old jj rats. This drug displaces bilirubin from its albumin binding sites in the circulation, shifting it into tissues including the brain. Marked latency prolongations (11-20%) and reduced amplitudes (20-64%) were observed in the different wave components of the VEP. These changes were evident as early as 2 h after injection of the drug and persisted thereafter for another 4 h. On the other hand, ERG changes (significant prolongation of wave b) became apparent in these animals only 6 h after SDM injection. These results suggest that, although some changes in the retina may occur after a massive entry of bilirubin into the nervous system, the primary damage in the visual pathway after bilirubin exposure is probably beyond the retina. PMID- 7478826 TI - Neurodevelopmental quotient of healthy term infants at 4 months and feeding practice: the role of long-chain polyunsaturated fatty acids. AB - A direct influence of dietary long-chain polyunsaturated fatty acids (LC-PUFA) on the developmental quotient (DQ) of the healthy term infant remains unexplored. To test this hypothesis, we designed a prospective study of three types of diet. Twenty-nine infants received a LC-PUFA-supplemented formula, 31 received a standard infant formula, and 30 infants were breast-fed exclusively. Neurodevelopmental response was measured by the Brunet-Lezine psychomotor development test at 4 mo. The fatty acid status was also assessed among three diet subgroups (59 subjects) at 4 mo. Formula-fed infants who received LC-PUFA supplementation scored significantly higher (p < 0.01) on the Brunet-Lezine scale than infants who received the standard formula. Breast-fed infants also performed better than those fed the standard formula. Arachidonic acid and docosahexaenoic acid levels in circulating lipids and erythrocyte phospholipids were higher among breast-fed infants and among the group fed the arachidonic- and docosahexaenoic acid-supplemented formula. These findings are suggestive that formula supplementation with one or both of these fatty acids can benefit term infants in neurodevelopmental performance. PMID- 7478827 TI - Children born small for gestational age: do they catch up? AB - Postnatal growth of 724 (423 premature, 301 full-term) small for gestational age infants (SGA, birth length less than the third length percentile (P3) for gestational age) was studied for the first 2 y of life. The study group consisted of all SGA infants who had been admitted over a period of 8 y at the Departments of Neonatology of three University Hospitals in The Netherlands with exclusion of infants with well defined causes for growth retardation, such as chromosomal disorders, syndromes, severe malformations, or complications during the neonatal period or later on. The aim of the study was to describe postnatal growth of SGA infants and to find predictive factors for catch-up growth > or = P3 during the first 2 y of life. The majority (around 85%) of the healthy SGA infants showed catch-up growth to a height > or = P3 during the first 2 y of life. The percentage of premature SGA infants with catch-up growth > or = P3 at 2 y of age (82.5%) was not significantly different from that of full-term SGA infants (87.5%). Birth length SDS was more sensitive than birth weight SDS in predicting catch-up > or = P3 in premature SGA infants. In contrast, birth weight SDS was the best predictor for catch-up > or = P3 in full-term SGA infants. Gestational age, multiple birth, and sex were not significantly associated with catch-up in height > or = P3.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7478828 TI - Hospitals' responses to universal infant hepatitis B vaccination recommendations. AB - OBJECTIVE: To assess the extent to which hospitals in a midwestern state with low acute hepatitis B virus (HBV) morbidity offered hepatitis B (Hep B) vaccine to all infants, whether offering infants Hep B vaccine was associated with hospital geographic location or size, as measured by the number of births, and how hospital staff resolved key programmatic issues. METHODS: The managers of hospital newborn nurseries (N = 110) were surveyed by mail. The written response rate was 72%; all of the nonresponders were interviewed by telephone. The outcome measured was the number of hospitals offering Hep B vaccine to all infants by geographic region and hospital size. RESULTS: Sixty-five percent of the hospitals routinely offered Hep B vaccine to all infants; these hospitals accounted for 80% of reported Wisconsin births. In univariate analysis, the decision to offer infants Hep B vaccine was associated with both hospital size and hospital location. After controlling for size, hospitals in the northeastern region were eight times more likely (relative risk, 8.21; 95% confidence interval, 1.30, 51.79) to offer infants Hep B vaccine than hospitals in the southeastern (referent) region. Regional differences in reported rates of acute HBV infection do not explain this finding, because morbidity in the northeastern region (1 per 100,000) is among the lowest in Wisconsin. Although more than 80% of hospitals with Hep B vaccination programs required written informed consent for vaccination, had standing orders for administering Hep B vaccine to infants whose mothers' hepatitis B surface antigen (HBsAg) test results were known, and had mechanisms to notify the infants' physicians that the infants had been vaccinated, only 38% had standing orders for testing mothers whose HBsAg test results were unknown. PMID- 7478829 TI - Impact of prenatal diagnosis on the birth prevalence of neural tube defects, Atlanta, 1990-1991. AB - OBJECTIVE: To determine the impact of prenatal diagnosis on the birth prevalence of neural tube defects (NTDs) in Atlanta during 1990 through 1991. METHODS: Live born and stillborn infants with NTDs who were at least 20 weeks' gestation were ascertained by the Metropolitan Atlanta Congenital Defects Program (MACDP), a population-based birth defects registry. Prenatally diagnosed NTD-affected pregnancies were ascertained from the four perinatal centers and the three genetic laboratories operating in Atlanta during 1990 through 1991. Fetal death certificates were also reviewed for potential cases. RESULTS: During 1990 through 1991, MACDP ascertained 59 NTD cases, for a birth prevalence of 0.77/1000 live births. During this period, an additional 28 NTD-affected pregnancies were detected prenatally and terminated before 20 weeks' gestation. The adjusted NTD rate during 1990 through 1991, which includes prenatally diagnosed cases, was 1.13/1000 live births. CONCLUSIONS: Prenatal diagnosis is making a substantial impact on the birth prevalence of NTDs in Atlanta. However, since NTD rates in Atlanta were 2 to 2.5 per 1000 live births in 1970, prenatal diagnosis and termination of pregnancy does not completely account for the declining rate of NTDs. PMID- 7478830 TI - A pacifier increases the risk of recurrent acute otitis media in children in day care centers. AB - OBJECTIVE: To follow up a previous retrospective analysis in which we found the use of a pacifier to be a risk factor for recurrent acute otitis media (AOM). METHOD: In the present prospective study, the occurrence of AOM and the use of a pacifier were recorded in 845 children attending day care centers during a 15 month period. RESULTS: More than three attacks of AOM occurred in 29.5% of the children younger than 2 years using pacifiers and in 20.6% of those not doing so (relative risk, 1.6; 95% confidence interval [CI], 0.6, 4.1); in children 2 to 3 years of age, the figures were 30.6% and 13.2%, respectively (relative risk, 2.9; 95% CI, 1.2, 7.3). Logistic modeling with adjustment for age and the duration of monitoring showed the occurrence of AOM to be associated with the time during which a pacifier was used. The use of a pacifier increased the annual incidence of AOM from 3.6 (95% CI, 2.5, 4.9) to 5.4 episodes (4.4, 6.6) in children younger than 2 years and from 1.9 (1.4, 2.5) to 2.7 (2.2, 3.3) in children 2 to 3 years of age. The population-attributable risk of AOM attacks due to the use of a pacifier was 176 attacks, ie, 459 to 635 attacks per year, in the youngest children and 69 attacks, ie, from 264 to 333 attacks per year, in those 2 to 3 years of age. It can be calculated that the use of a pacifier was responsible for 25% of the attacks in children younger than 3 years. Breastfeeding, parental smoking, thumb sucking, using a nursing bottle, and the social class of the family failed to show such strong associations with the occurrence of AOM. CONCLUSION: We conclude that the use of a pacifier is a significant risk factor for recurrent AOM and suggest that pacifiers should be used only during the first 10 months of life, when need for sucking is strongest, and AOM is uncommon. PMID- 7478831 TI - Beliefs and practices regarding childhood vaccination among urban pediatric providers in New Jersey. AB - BACKGROUND: In 1991, the fourth largest measles outbreak in the nation (824 cases) occurred in the Jersey City, New Jersey area. Data from a subsequent intervention trial in Jersey City demonstrated that vaccinations were more likely to be delayed for children who had received care from private rather than public clinic providers. In addition, failure to administer multiple indicated vaccines at a single visit was associated with vaccination delay, and reluctance to administer multiple vaccines was more common among private providers. These findings prompted an investigation of vaccination beliefs and practices among urban pediatric providers. METHODS: A telephone survey of vaccination beliefs and practices was administered to all pediatric providers in both private and public clinics in the Paterson and Jersey City areas. RESULTS: Private providers were less likely than public clinic providers to consider vaccinating children during emergency room visits (relative risk [RR] = 2.2; 95% confidence interval [CI] = 1.2-4.2) or hospital admissions (RR = 13.2; 95% CI = 1.9-92.7) and less likely to believe that all recommended vaccine doses should be administered simultaneously (RR = infinite; lower 95% confidence limit = 3.0). Private providers were less likely to consider administering live-virus vaccines to children with minor acute illnesses and low-grade fever (RR = 2.2; 95% CI = 1.2-3.8) or killed-virus vaccines to children with minor acute illnesses without fever (RR = 3.4; 95% CI = 1.4-8.5) or with low-grade fever (RR = 2.2; 95% CI = 1.2-3.9). Private providers were more likely to believe that multiple injections should be avoided because of potential psychological and physical trauma to the child (RR = 4.0; 95% CI = 1.3 12.3). CONCLUSIONS: Adherence to Standards for Pediatric Immunization Practices by pediatric providers could improve vaccine coverage rates among urban children. PMID- 7478832 TI - A community-based survey of infant sleep position. Pediatric Research in Office Practices Network. AB - OBJECTIVE: To determine prevalent infant sleep positions before and after the American Academy of Pediatrics position statement of 1992 and to identify determinants of sleep position. METHOD: DESIGN: cross-sectional survey. SETTING: private and hospital-sponsored general pediatric offices. PARTICIPANTS: parents of infants younger than 7 months of age. RESULTS: Eight hundred fifty-two care givers completed surveys during the 5-week study. Fifty-four percent of the study infants were put to sleep in the prone position. In 416 families with more than one child, however, 75% of the youngest siblings had been put to sleep in the prone position at the same age. Gender, race, family income, maternal smoking, and birth weight were not associated with choice of sleep position. CONCLUSIONS: There has been a change in infant sleep positioning in the desired direction since the American Academy of Pediatrics statement. However, 54% of the study infants were still being put to sleep prone. PMID- 7478833 TI - Controlled trial of dexamethasone in neonatal chronic lung disease: a 3-year follow-up. AB - OBJECTIVE: We compared the effects of a policy of neonatal steroid administration versus placebo for babies chronically dependent on supplemental oxygen in terms of long-term health and development, judged at 3 years of age. DESIGN: Double blind randomized controlled trial. SETTING: Thirty-one centers in the United Kingdom, Ireland, Belgium, Germany, Canada, and the United States. PATIENTS: Babies who were chronically dependent on supplemental oxygen between 2 and 12 weeks of age were recruited to the trial between 1986 and 1989. Sixty-two children were known to have died, 23 before discharge from the hospital and 10 afterward in the active group, compared with 25 and 4, respectively, in the placebo group. Information was available for 209 of the 212 eligible for follow up (99%). INTERVENTIONS: A 1-week course of active dexamethasone phosphate 0.6 mg/kg/d (dexamethasone base 0.5 mg/kg/d) or saline placebo was given intravenously (or orally, if no intravenous line). There was an option to give a second tapering 9-day course if relapse occurred after initial improvement. OUTCOME MEASURES: Information about respiratory problems, growth, neurodevelopment and disability, infection, and health service use when the children were 3 years old was ascertained from questionnaires to general practitioners, health visitors, and parents (and occasionally pediatricians). RESULTS: About half the children in both groups had been admitted to the hospital for respiratory problems, with more in the active than the placebo group having at least five outpatient consultations for these problems over the 3 years. Overall, the children were below average in height, weight, and head circumference. About one fifth had cerebral palsy, 8% some visual loss, and 16% hearing loss; 18% needed or were anticipated to need special schooling. There were no clear differences between the randomized groups. These overall conclusions were not altered by any of the prespecified secondary analyses. CONCLUSIONS: Despite early benefits, there were no clear effects at 3 years of age. As 40% of the placebo group eventually received open steroids, even a trial of this size has limited statistical power to detect a moderate effect of the policy. Regardless of random allocation, overall morbidity was high, confirming that babies with protracted dependence on supplemental oxygen are at high risk of childhood disability and poor health. PMID- 7478834 TI - Survivors of extracorporeal membrane oxygenation at 1 year of age: the relationship of primary diagnosis with health and neurodevelopmental sequelae. AB - OBJECTIVE: Although extracorporeal membrane oxygenation (ECMO) has been responsible for the improved survival of infants with cardiorespiratory failure, its use over the last decade has raised concern as to the health of the survivors and the severity of neurodevelopmental sequelae. Though infants meeting ECMO criteria have a variety of reasons prompting the use of this therapy, most studies to date have simply reported outcome on the entire population that has survived without regard to the original nature of the child's illness. The purpose of this study was to determine the type and extent of health-related problems and neurodevelopmental sequelae in infants requiring ECMO therapy and the association of these findings with the infants' primary diagnosis. METHODS: Eighty-two neonates required ECMO therapy between May 1990 and December 1993. The most common diagnosis prompting ECMO therapy included 26% with meconium aspiration syndrome, 34% with congenital diaphragmatic hernia (CDH), 16% with persistence of the fetal circulation, and 9% with sepsis. Information concerning the hospital course was obtained through chart review, and the infants were seen at 6 and 12 months of age for medical and neurodevelopmental follow-up. Data were analyzed using descriptive statistics and Fisher's exact test, t-tests, and analysis of variance where appropriate. Assessment of hospital course and discharge data focused on the four main diagnostic groups, whereas follow-up data were further limited to the two most frequently encountered groups (meconium aspiration syndrome and CDH). RESULTS: Overall survival was 79%. Significant differences in survival were noted based on primary diagnostic category. Those with CDH fared the worst, with an overall survival rate of 68% and a more complicated hospital course with a longer duration of ECMO. At discharge, the CDH group demonstrated a greater incidence of bronchopulmonary dysplasia, gastroesophageal reflux, feeding dysfunction, and hypotonia. No significant differences were noted in the incidence of intraventricular hemorrhage, cerebral infarction, extra-axial fluid collection, or seizures. Hearing loss was uncommon. During the first year of life, although no differences were noted in growth rate, infants in the CDH group continued to experience a higher incidence of gastroesophageal reflux (43%) and feeding dysfunction, with 36% of this group requiring tube feedings for nourishment. Although 40% of the entire ECMO population was diagnosed with bronchopulmonary dysplasia before initial discharge, by 1 year of age, 50% of those with CDH versus 17% of those with meconium aspiration syndrome continued to be clinically symptomatic. Although the ECMO population as a whole scored in the normal range developmentally, CDH infants had significantly lower motor and slightly lower cognitive scores at 1 year of age. Despite finding abnormal muscle tone in a high percentage of the entire ECMO population at discharge, most demonstrated resolution by 1 year of age. Of the CDH infants, however, 75% continued to evidence some degree of hypotonicity, which affected acquisition and quality of gross motor skills. CONCLUSION: Despite the impact that ECMO has had on the survival of infants with severe respiratory failure, the efficacy of ECMO cannot be assessed accurately without an analysis of the extent and morbidity in the surviving population. Most centers are reporting relatively low morbidity for the entire ECMO population. However, upon separating this population into primary diagnostic categories, we found that the CDH population encountered a greater number of neurodevelopmental, respiratory, and feeding abnormalities during the first year of life. The reasons for these differences are unclear but may be related to the severity of the primary illness itself or the variables associated with prolonged ECMO therapy. Stratifying outcome by primary diagnosis gives the health care provider more information to improve PMID- 7478835 TI - Elevated serum lactate correlates with intracranial hemorrhage in neonates treated with extracorporeal life support. AB - OBJECTIVES: To correlate the initial and maximal lactate levels with the occurrence of intracranial hemorrhage (ICH) and survival in patients treated with extracorporeal life support (ECLS). DESIGN: Retrospective chart review. SETTING: Pediatric intensive care unit. PATIENTS: Eighty-two neonatal patients placed on ECLS for respiratory failure due to sepsis, meconium aspiration, or persistent pulmonary hypertension of the newborn. MEASUREMENTS: The initial lactate level measured within 6 hours of initiating ECLS and the maximal lactate level measured throughout the ECLS course were collected. Lactate levels were described as mean lactate +/- SE (mM). Head ultrasound reports and survival were reviewed. Platelet counts and activated clotting times (ACTs) were examined. RESULTS: The mean initial and maximal lactate levels were higher in ECLS patients who developed ICH (initial: 10 +/- 1.7 mM vs 6.4 +/- 0.8 mM, p = .05 and maximal: 12.4 +/- 2.5 mM vs 7.9 +/- 0.8 mM, p = .04). Initial and maximal lactate levels were also elevated in nonsurvivors (initial: 11.7 +/- 3 mM vs 6.4 +/- 0.7 mM, p = .01 and maximal: 14.8 +/- 3.3 mM vs 7.8 +/- 0.8 mM, P < .01). Platelet counts and ACT did not differ in patients with and without ICH. CONCLUSIONS: Lactate is a useful marker for the development of ICH in ECLS patients. In addition, elevated lactates during ECLS identify a subgroup of patients with poor outcome. Prospective studies are needed to determine whether the incorporation of this information into pre-ECLS and ECLS management will decrease the occurrence of ICH and improve survival. PMID- 7478836 TI - Score for neonatal acute physiology: validation in three Kaiser Permanente neonatal intensive care units. AB - BACKGROUND: Measurement of the severity of illness is a research area of growing importance in neonatal intensive care. Most severity of illness scales have been developed in tertiary care settings. Their applicability in community neonatal intensive care units has not been tested. OBJECTIVES: Our goal was to assess the operational characteristics of the score for neonatal acute physiology (SNAP): the relationship to birth weight, the length of total hospital stay, and in hospital mortality. METHODS: We assigned SNAP scores prospectively to all inborn admissions at three community neonatal intensive care units during an 11-month period. Data on other neonatal predictors (eg, birth weight and the presence of congenital heart disease) were also collected. We measured in-hospital mortality, the experience of interhospital transport to a higher level of care, and total hospital stay. RESULTS: We found that the SNAP's relationship to birth weight was similar to previous reports. The SNAP's perinatal extension is a reliable predictor of newborn in-hospital mortality, with an area under the receiver operator characteristic curve of 0.95. The SNAP is also a good predictor of total hospital length of stay, whether by itself (by which it can explain 31% of the total stay) or in combination with other variables. Its predictive ability is better among infants of low birth weight (<2500 g) than among those of normal birth weight (> or = 2500 g). The SNAP's predictive power was most limited among infants admitted to rule out sepsis. The predictive ability of a model containing birth weight, the SNAP, and transport status was not improved by the inclusion of two major diagnostic categories, the presence of congenital heart disease or complex illness. CONCLUSION: Although it has definite limitations among infants who weight 2500 g or more, the SNAP is a potent tool for outcomes research. Modification of some of its parameters could result in a multifunctional scale suitable for use with all birth weights. PMID- 7478837 TI - Effectiveness of individualized developmental care for low-risk preterm infants: behavioral and electrophysiologic evidence. AB - OBJECTIVE: We assessed the effectiveness of individualized developmental support in the special care nursery for low-risk preterm infants. SETTING: A university affiliated teaching hospital. PARTICIPANTS: Twelve healthy full-term infants, and 24 low-risk preterm infants randomly assigned to a control or an experimental group. DESIGN: The preterm control group received standard care and the preterm experimental group received individualized developmental care at the same special care nursery. OUTCOME MEASURES: Medical, behavioral (Assessment of Preterm Infants' Behavior and Prechtl's Neurological Examination of the Full-Term Newborn Infant), and electrophysiologic outcome (using quantitative electroencephalography with topographic mapping) of all three groups was assessed 2 weeks after the expected due date. RESULTS: No between- or among-group medical differences were seen for this low-risk, healthy sample. The preterm experimental group showed behavioral and electrophysiologic performances comparable to those of the full-term group, whereas the preterm control group performed significantly less well. Behavioral measures suggested significantly poorer attentional functioning for the preterm control group. Electrophysiologic results implicated the frontal lobe. CONCLUSIONS: Individualized developmental intervention supports neurobehavioral functioning as measured at 2 weeks post-term. It appears to prevent frontal lobe and attentional difficulties in the newborn period, the possible causes of behavioral and scholastic disabilities often seen in low-risk preterm infants at later ages. PMID- 7478838 TI - Psychological changes in parents eight months after the loss of an infant from stillbirth, neonatal death, or sudden infant death syndrome--a longitudinal study. AB - OBJECTIVE: We proposed to measure part of the natural history of grief by determining the changes in the psychological symptoms experienced by bereaved parents over the 8 months after the loss of an infant from sudden infant death syndrome (SIDS), neonatal death (NND), or stillbirth (SB). Parents were interviewed twice, at 2 and 8 months after the loss. METHODOLOGY: A total of 220 bereaved families (45 SIDS, 93 NND, and 82 SB) were compared with 226 control families who had a live born child. Comparison was based on responses to a standardized measure of anxiety and depression (Delusions-Symptoms-States Inventory). RESULTS: For separate cross-sectional comparison at both 2 and 8 months, significant differences were noted in the frequency of maternal symptoms of anxiety and depression between bereaved and control groups (P < .001). Such differences were present for paternal anxiety and depression at 2 months, but not 8 months. A second series of analysis examined longitudinal changes in symptom frequency between 2 and 8 months for each bereaved group. For mothers, the changes were significant for anxiety and depression: SIDS and NND (P < .001), SB (P < .01). For fathers, the changes for anxiety and depression in SIDS were P < .01; NND, P < .05 for anxiety and P < .01 per depression; changes were not significant for SB. At 2 months, relative risks for symptoms of maternal anxiety were significant for all three bereaved groups: SIDS, 22.4; NND, 5.4; and SB, 5.1. Comparable significant figures at 8 months were: SIDS, 5.5; NND, 3.9; and SB, 3.0, respectively. For depression the results for 2 months were: SIDS, 8.6; NND, 5.9; and SB, 6.7 (all significant) while at 8 months the results were: SIDS, 5.1; NND, 3.8; and SB, 2.4 (SB group not significant). For fathers the relevant risks were generally lower. At 2 months, anxiety levels were higher than controls in all three groups, and for depression in the SIDS and SB groups. At 8 months, significant results persisted only in the SB group. CONCLUSION: These data indicate that bereaved parents have a marked reduction in the symptoms of mental illness over the first 8 months after the loss. Although the changes over time are significant for both mothers and fathers, mothers at 8 months still demonstrated higher levels of anxiety and depression when compared with controls. These levels of symptoms are far less evident for fathers at 8 months. PMID- 7478839 TI - Neonatal thrombosis: report of a prospective Canadian and international registry. AB - OBJECTIVE: We sought to obtain representative data on the risk factors, diagnosis, current management, and short-term outcome of neonatal thrombosis. RESEARCH DESIGN: A case registry was established at McMaster University. Standardized questionnaires were mailed to collaborators at participating centers every 4 to 6 months. SETTING: Eighty-five level III and modified level II neonatal units in North America, Europe, and Australia were invited to join the registry. PATIENTS: Eligible infants were born between January 1990 and June 1993. Large-vessel thrombosis was diagnosed during the first month of life or up to 44 weeks post-conception after premature birth. The clinical impression of thrombotic vessel obstruction was confirmed using at least one imaging technique. RESULTS: Physicians in 64 centers expressed their willingness to participate. A total of 97 cases (excluding stroke) were registered from 29 centers. Spontaneous renal venous thrombosis (n = 21) was diagnosed at a median age of 2 days. The other venous (n = 39), arterial (n = 33), and mixed (n = 4) thromboses presented later; 89% of them were associated with an intravascular catheter and 29% with systemic infection. Doppler ultrasonography was the definitive diagnostic test in 68% of cases; contrast angiography was performed infrequently (14%). A third of all patients (but 62% of infants with renal venous thrombosis) received supportive therapy only. Thrombolytic agents were prescribed for 28% of catheter associated venous thromboses and 30% of all arterial thromboses. The remainder of the patients were given heparin. Most patients (82%) survived to hospital discharge. Mortality rates were highest among infants with aortic thrombosis or central venous line-associated thrombosis affecting the right atrium or the superior vena cava (33%). CONCLUSIONS: Neonatal thrombosis is diagnosed fairly rarely. With the exception of spontaneous renal venous thrombosis, almost all cases are associated with indwelling catheters. Doppler ultrasound techniques are the most popular means of confirming the diagnosis in virtually all centers. Treatment varies greatly among different centers, probably because of the lack of scientific evidence about the optimum management of affected infants. PMID- 7478840 TI - Adverse factors associated with forced sex among southern adolescent girls. AB - OBJECTIVE: To identify adverse behavioral and psychological factors associated with forced sex experiences of adolescent girls compared with their sexually active counterparts. METHOD: An anonymous self-report survey examining an array of psychosocial items, to which 3124 grade 8 and grade 10 female students responded. RESULTS: Sexually abused girls were more likely to have been pregnant, to have initiated sexual intercourse at a younger age, to indicate illegal drug use, to have feelings of depression, to express more frequent suicidal ideation, and to have been physically abused. CONCLUSIONS: Behaviors such as gateway drug use, truancy, binge drinking of alcohol, and participation in violent episodes that were previously identified as indicators of sexual abuse did not distinguish between sexually active adolescents and those who had been sexually abused. Physicians should consider carefully a structured series of questions relating to behaviors as one approach in determining the risk of sexual abuse. Positive responses to young sexual initiation, pregnancy, illegal drug use, negative mental health states, and evidence of physical abuse are potential markers of sexual abuse in adolescent female clients. PMID- 7478841 TI - Severe dog bites in children. AB - OBJECTIVE: We evaluated children less than 16 years of age who had dog bite injuries that resulted in hospitalization or death to determine the typical characteristics of the children, the dogs, and the injuries suffered. DESIGN: Retrospective chart review. SETTING: Three large city hospitals including Harborview Medical Center, Seattle, Washington; Children's Mercy Hospital, Kansas City, Missouri; and Mary Bridge Hospital, Tacoma, Washington. METHODS: Charts were reviewed for patient demographic data and canine data. Hospitalization data included total length of stay, need for intensive care, Injury Severity Score, the nature and extent of the injuries, procedures performed, complications, and outcome. RESULTS: Forty cases were reviewed. Most children were boys (60%) and were white (87%). The median age was 50 months. There were three deaths. Most dogs were medium-sized or large breeds and were familiar to the victim. The average hospital stay was 6 days (SD = 5), and 12 (30%) patients required a stay in the intensive care unit. Injuries to the face, head, and neck area were most common (82%). Major surgical procedures included craniotomy, exploration of the neck or abdomen, ocular procedures, and repair of fractures. CONCLUSIONS: Severe dog bites in children occur most frequently in those younger than 5 years old and involve the head and neck. Large dogs that are familiar to the child are usually involved. Young children should be closely supervised when around any dog. PMID- 7478843 TI - A new device for pleural drainage in newborn infants. AB - OBJECTIVE: We tested a chest tube device and insertion technique designed to reduce complications and expedite effective thoracostomy tube placement. METHODOLOGY: This is a case series study. We placed chest tubes using this device in 25 infants ranging in weight from 425 to 3500 g. This investigation was performed in a regional referral center neonatal intensive care unit. RESULTS: Using this device, chest tube insertion was safe and effective. No complications were identified. CONCLUSION: We believe percutaneous placement of small-bore pigtail catheters using this technique represents a useful alternative to the standard approach of thoracostomy tube placement in small infants. SYNOPSIS: To reduce complications during thoracostomy tube placement and to expedite tube insertion, we modified a chest tube device for use in premature infants. This technique and device facilitates quick and accurate placement. We placed chest tubes using this device in 25 infants ranging from 425 to 3500 g. Using this device, chest tube insertion is quick, easy, and safe. No complications were identified. We believe percutaneous placement of small-bore pigtail catheters using this technique represents a useful alternative to the standard approaches of thoracostomy tube placement in small infants. PMID- 7478842 TI - Effect of urine latex agglutination tests on the treatment of children at risk for invasive bacterial infection. AB - BACKGROUND: Identifying febrile children with invasive bacterial infection is difficult in the absence of telltale physical findings. Urine latex agglutination (ULA) tests have been used for rapid, on-site identification of such children. OBJECTIVES: To study the performance of ULA tests in identifying children with Haemophilus influenzae, Streptococcus pneumoniae, and group B streptococcus infection and to examine how the results of ULA tests affect patient treatment. DESIGN: Retrospective review. SETTING: Urban children's hospital. PATIENTS: All emergency department and hospital patients tested by ULA in 1990, excluding patients in the neonatal units. RESULTS: Of 1629 patients, 36 had positive tests (20 H influenzae, 5 S pneumoniae, and 11 group B streptococcus). Thirteen of these were false positive based on culture results. Although ULA tests demonstrated excellent specificity, their sensitivity was poor. Positive predictive values for bacteremia ranged from 0.346 to 0.600, and negative predictive values ranged from 0.972 to 0.997. The decision to treat with antibiotics was made before ULA test results were available in 19 (53%) of the 36 patients with positive test results. Of 1593 patients with negative test results, 1211 (76%) were admitted to the hospital, and approximately 81% were empirically treated with parenteral antibiotics. CONCLUSIONS: In clinical practice, treatment and disposition decisions are frequently made without regard to ULA test results. PMID- 7478845 TI - Analysis of homeopathic treatment of childhood diarrhea. PMID- 7478844 TI - Increased incidence of severe breastfeeding malnutrition and hypernatremia in a metropolitan area. AB - OBJECTIVE: To identify common characteristics among infants with breastfeeding malnutrition in a region with an increasing incidence of breastfeeding malnutrition. DESIGN: Retrospective case series. SETTING: A 361-bed regional tertiary care children's hospital in a 1.7 million population metropolitan area. PATIENTS: CASE SERIES: five infants with severe breastfeeding malnutrition and hypernatremia admitted to a tertiary care children's hospital over a 5-month period. Retrospective case review: 166 infants admitted between 1990 and 1994 with the diagnosis of dehydration, hypernatremia, or malnutrition. MAIN OUTCOME MEASURES: Maternal characteristics, age at presentation, percent loss from birth weight, serum sodium, average age at birth hospital discharge, neurologic, or cardiovascular complications. RESULTS: Five infants were admitted to a children's hospital over a 5-month period with severe breastfeeding malnutrition and hypernatremia. The average weight loss at time of readmission was 23% (+/- 8%) from birth weight. The average presenting sodium was 186 +/- 19 mmol/L. Three suffered significant complications. From 1990 through 1994, there was a statistically significant (P < .05) annual increase in the number of infants admitted with breastfeeding malnutrition and hypernatremia. CONCLUSIONS: While breastfeeding malnutrition and hypernatremia is not a new problem, this cluster of infants represents an increase in frequency and severity of the problem and could be a consequence of several factors, including inadequate parent education about breastfeeding problems and inadequate strategies for infant follow-up. PMID- 7478846 TI - Bright futures. PMID- 7478847 TI - Early discharge alert. PMID- 7478848 TI - Children's Medicaid entitlement: what have we got to lose? PMID- 7478850 TI - Parental leave for residents and pediatric training programs. American Academy of Pediatrics Committee on Early Childhood, Adoption, and Dependent Care Resident Section. PMID- 7478849 TI - New developments with pertussis vaccines. PMID- 7478851 TI - Perinatal care at the threshold of viability. American Academy of Pediatrics Committee on Fetus and Newborn. American College of Obstetricians and Gynecologists Committee on Obstetric Practice. AB - The survival rate for infants at the threshold of viability has been improving. However, there are insufficient data regarding the cost(s) of initial and ongoing care of these infants and the long-term outcome of survivors. Furthermore, there has been little study of the impact of obstetric management on the survival rates of extremely low birth weight infants and on long-term morbidities. Continued research on these issues is imperative, and physicians need to remain informed of changing statistics. PMID- 7478852 TI - Human milk, breastfeeding, and transmission of human immunodeficiency virus in the United States. American Academy of Pediatrics Committee on Pediatric AIDS. PMID- 7478853 TI - In-hospital mortality for surgical repair of congenital heart defects: preliminary observations of variation by hospital caseload. PMID- 7478854 TI - Informed consent, parental permission, and assent in pediatric practice. PMID- 7478855 TI - Ultrasound screening for congenital dysplasia of the hips. PMID- 7478856 TI - Ear temperatures. PMID- 7478857 TI - Esophageal detector device in pediatric tracheal intubation. PMID- 7478859 TI - Temporal factors for vibrotactile letter reading. AB - Using an apparent-motion mode, the psychophysical effect of apparent movement was introduced for tactile letter reading. This mode reduces time for the tracing mode. Five temporal parameters were examined for application to actual communication. 4 subjects recognized words composed of 3 to 7 Japanese letters in about 5 seconds. PMID- 7478858 TI - Human analog tests of the sixth stage of object permanence. AB - Two adult cats were tested on multiple invisible displacement. A dowel was established as a secondary reinforcer and hidden in a manner similar to that used to assess the culmination of sensorimotor intelligence in human infants. Three other cats were tested on single invisible displacement, a simpler version of the task. For human infants, this task is used to assess the beginning of mental representation in the sixth and last stage of sensorimotor intelligence. The cats' searches on these tasks were consistent with representation of an unsensed object and fully developed sensorimotor intelligence. PMID- 7478860 TI - Influences on the Mantel-Haenszel chi-square in detection of differential item functioning under Rasch conditions. AB - The Mantel-Haenszel chi-square (chi 2 MH) is widely used to detect differential item functioning (item bias) between ethnic and gender-based subgroups on educational and psychological tests. The empirical behavior of chi 2 MH has been incompletely understood; previous research is inconclusive. The present simulation study explored the effects of sample size, number of items, and trait distributions on the power of chi 2 MH to detect modeled differential item functioning. A significant effect was obtained for sample size with unacceptably low power for 250 subjects each in the focal and reference groups. The discussion supports the 1990 recommendations of Swaminathan and Rogers, opposes the 1993 view of Zieky that a sample size of 250 for each group is adequate. PMID- 7478861 TI - Body image and social attitude in growth-hormone-deficient adults. AB - This study was done to evaluate the perception of body image and psychosocial attitudes in a sample of 8 male adult subjects with growth-hormone deficit before and after six months of therapy. Before treatment the subjects tended to underestimate body size and were characterized by low self-esteem, a closed attitude towards social relationships, a pessimistic attitude with a tendency towards depression, and a strong sense of detachment from the outside world. After the treatment, even if the subjects continued to underestimate body size, we observed an improvement in terms of emotional control and well-being, accompanied by lower scores on anxiety. An integration of a psychological support in the GH therapy could help the patients to overcome problems of bodily and social acceptance. PMID- 7478862 TI - Environmental changes affect sleep patterns: a case study. AB - While at a state institution for people with mental retardation Peter averaged 5.30 hr. sleep per night. At his community home he averaged 7.87 hr. per night, a 48% gain. The responsive environment of his new home engaged Peter in positive social activities which may have led to decreased stress and increased personal comfort, enabling him to sleep longer. PMID- 7478863 TI - Effects of color on children's naming of pictures. AB - Two experiments assessed possible effects of memory for the typical colors of objects on children's naming of pictures. Children, 5 to 10 yr. old (N = 54 per experiment), named pictures of familiar objects in uncolored, typically colored (red for apple), and atypically colored (blue for apple) versions. Presentation of the three types of pictures was mixed in Exp. 1 and blocked in Exp. 2. With mixed presentation, reaction times for naming were unaffected by typical colors but slowed by atypical colors. With blocked presentation, naming was speeded by typical colors, but only for younger children, and was unaffected by atypical colors. Implications are discussed for understanding the processes by which objects are identified and named and the development of knowledge regarding the color of objects. PMID- 7478864 TI - Fast finger extensions are slower in stutterers than in nonstutterers. AB - 10 adult stutterers and 10 nonstutterers were required to extend as fast as possible their right and left index fingers under isometric conditions in response to an auditory signal. Force developed during finger extension was measured continuously during each trial. From these force records reaction time, contraction time, and peak force were measured. For stutterers, contraction times were longer (about 36 msec.) than for nonstutterers. Possible reasons for this result are discussed. PMID- 7478865 TI - Mechanical perturbation of jaw movements during speech: effects on articulation and phonation. AB - 12 subjects uttered the testword/papapas/repeatedly with three different speech rates and two stress patterns. On 17% randomly chosen trials, a mechanical load was applied unpredictably to the jaw in the direction of the opening movement. Load onset was triggered by the start of the first phonation. Analysis showed that the opening and closing displacements of the jaw movement in the first syllable were not influenced significantly by the perturbation. The load application prolonged the duration of the jaw movement in unstressed syllables but not in stressed syllables. Further, the mechanical perturbation of the jaw led to increased duration of phonation in unstressed syllables only, the effect for duration of phonation being greater at higher speech rates. These results demonstrate a coupling between articulation and phonation. PMID- 7478866 TI - Validity and reliability of a Mainstreaming Attitude Inventory for child-care providers. PMID- 7478868 TI - Effect of task demand and stimulus relevance on preattentive processing. AB - Evidence of cognitively controlled processes influencing preattentive processes is provided. In the first experiment, preattentive processing associated with a discrimination task was compared with that associated with a reaction task. An increase in processing time due to preattentive processes was only found for the discrimination task. In the second experiment, the effects of different amounts of stimulus relevance on preattentive processing were investigated. Stimulus relevance was varied by changing the spatial and temporal distance in cognitively controlled processing. The relevance of the stimulus was proportional to the amount of preattentive processing. It was concluded that the influence of cognitively controlled processing on preattentive processing is moderated by task demand and stimulus relevance. These results are based on observations concerning processing time. PMID- 7478867 TI - Relations of oral imagery to thought disorder in subjects with frequent nightmares. AB - Oral imagery was examined in the Rorschach protocols of 60 female subjects, 30 who experienced at least one nightmare per week and 30 who had no more than one nightmare in the past year. Mean number of oral responses did not differ between the groups. In subjects with nightmares, Rorschach orality was significantly related to many personality measures, including state and trait anxiety, Beck Depression Inventory scores, five subscales of a schizotypy inventory, the Rorschach penetration score, and two measures of pathological thinking on the Rorschach. In the 30 subjects without nightmares, the number of significant correlations did not exceed that expected by chance. The results are discussed within the context of empirical and theoretical reports about the links between the oral personality and pathological behavior. PMID- 7478869 TI - Physiological sensations of initial smoking in the development of regular smoking behavior. AB - This study examined the relationships between adolescents' physiological sensations of smoking during initiation and early experience. For a national sample of a birth cohort of 2,043 adolescents, ages 15 to 22 years at the follow up, variables of interest included measures of smoking behavior and physiological sensations reported from the initial smoking experience. Analysis showed that adolescents experimenting with smoking were more likely to become regular smokers over three years if they indicated that they felt relaxed, felt dizzy, did not feel sick, and did not cough during the initial smoking experience. Antismoking interventions may impede the transition to regular smoking by helping adolescents interpret the physiological sensations as negative and unhealthy. PMID- 7478871 TI - Asymmetries in ear movements and eyebrow raising in men and women and right- and left-handers. AB - The abilities to move ears and eyebrows were examined in 442 subjects (204 men, 238 women) categorized as right-handed (n = 382) and left-handed (n = 60, including mixed and ambidextrous-handed subjects). Approximately 22% could move one or the other ear and about 18% could move both ears simultaneously, but significantly more men could move both ears simultaneously. Significantly more men than women were able to move both the left and right eyebrow and the left ear. No differences were observed between right- and left-handers. Significant contingency correlations were observed between raising eyebrows and moving ears. Results are discussed with reference to a possible left ear-right hemisphere advantage for localising environmental sounds, primitive ear-moving abilities no longer functional in modern humans, and epiphenomenal by-products of other adaptive sex differences. PMID- 7478870 TI - Optimizing movement speed for acquisition across levels of intelligence. AB - Three movement speeds (400, 800, and 1200 msec.) were examined to ascertain the optimal condition under which differing intellectual and developmental groups might acquire an arm-movement task. Parallel acquisition was observed only at the two slower speeds (800 and 1200 msec.), hence the faster of these (800 msec.) was deemed to be most optimal for expanded investigation. PMID- 7478872 TI - Field dependence-independence of male and female Spanish athletes. AB - We examined the field dependence-independence of Spanish athletes, 293 men and women (aged 18 to 23 years) active in individual and team sports, and nonathletes. Subjects were tested on the Group Embedded Figures Test. Analysis indicated that men active in team sports were more field-dependent than those non involved in sports, and that team-sport women exhibited a higher field-dependence than individual-sport athletes or nonathletes of both sexes. Our data reflect a significant interaction between type of sport and sex. PMID- 7478873 TI - On the psychophysics of arm-positioning movements. AB - The basis for the relationship between physical stimulus and magnitude of psychological response in angular movements of the upper limbs was investigated. Participants compared movements to a standard location with movements to comparison locations and movements of a standard distance with movements of comparison distances. The results add to the controversy surrounding the explanations for the differences in psychological response magnitude of movements to locations and movements across distances. Data obtained from movements to locations and movements across distances indicated similar Weber ratios but different patterns for the just noticeable differences. The data of this and previous research suggest that the application of a prothetic continuum to arm movements is inappropriate. An alternative explanation of the differences in the perception of movements to locations and movements across distances is offered. PMID- 7478874 TI - Imagery and physical practice in the acquisition of gross motor timing of coincidence by adolescents with mild mental retardation. AB - The purpose of this study was to examine the effect of imagery practice in conjunction with physical practice on the performance of anticipating a coincidence (striking) by adolescents with mild mental retardation. 32 adolescents were randomly assigned to either a physical practice plus imagery practice group or a physical practice only group. Subjects in the physical practice plus imagery practice group were asked to image the task before physically performing it, while subjects in the physical practice only group did not image the task. Subjects performed 20 trials per day for five consecutive sessions (days). The physical practice plus imagery group performed with significantly greater accuracy and less variability than subjects in the physical practice only group, and subjects regardless of group affiliation were able to reduce error and variability over the study. These results support the use of imagery practice in conjunction with physical practice when performing a relevant anticipation of coincidence (striking) task as well as an aid in reducing performance variability. Based on the increased amount of cognitive-symbolic element in the striking task as opposed to tasks used in previous studies, evidence is presented for support of the notion that imagery facilitates motor performance to the extent that cognitive-symbolic elements are present. PMID- 7478875 TI - Personal goals of young social drop-outs. AB - To examine the extent to which personal goals and their appraisals are associated with problems in socialization, 20 young 'social drop-outs' (15 men, 5 women) and 21 students from a vocational school (20 men, 1 woman) were interviewed about their personal goals, related views of internality, externality, and likelihood of accomplishing goals. Analysis indicated that young adults who showed problems in socialization mentioned less frequently personal goals related to future education and housing than did the control group. Second, social drop-outs held more external and less internal views and were less optimistic about accomplishing personal goals than was the control group. PMID- 7478876 TI - Stimulus-driven or autonomous shift or attention? AB - Hsieh and Allport studied shifts of attention in semantic space, using a semantic monitoring task based on rapid, serial, visually presented sequences of words. They reported that following a shift of criterion, accuracy of semantic monitoring dropped abruptly to a low level, then gradually recovered to reach the preshift level over successive stimuli in the sequence. They further examined the nature of the recovery of accuracy following a shift of criterion. Despite the striking results they obtained, some problems of their design remained. Hence, the current research replicated Hsieh and Allport's experiments with some modifications and showed that a shift of semantic criterion in a rapidly presented sequence indeed appears to be stimulus-driven rather than an autonomous process. PMID- 7478877 TI - Test-retest reliability of the survey form of the Leyton Obsessional Inventory Child Version. AB - The test-retest reliability of the survey form of the Leyton Obsessional Inventory-Child Version was examined over a 2-wk. interval. Three age groups of children participated in the study, 8- to 10-year-olds, 11- to 13-year-olds, and 14-to 16-year-olds (N = 106). For the total Obsessive scores, the test-retest Pearson correlation coefficients for the three age groups were .51, .75, and .83, respectively, which suggest that the temporal stability of the instrument is a function of age. PMID- 7478878 TI - Intermittent vision and discrete manual aiming. AB - This study was designed to assess how the precision requirements of discrete aiming movements affect the utility of brief visual samples provided during execution of movement. Subjects pointed with a hand-held stylus to targets with indices of difficulty of 3, 4, 5, and 6 bits with full vision, no vision, and in conditions in which 20-msec. visual samples were provided every 80, 140, or 200 msec. While intermittent vision required slightly longer movement times for targets with a high index of difficulty, subjects' accuracy was similar to the full-vision situation. Moreover, with intermittent vision, the movement trajectories resembled the full-vision and not the no-vision situation. It would appear that brief visual samples of the movement environment are sufficient for reasonably precise closed-loop control. PMID- 7478879 TI - Estimates of reliabilities for the Teacher Integration Attitude Questionnaire. PMID- 7478880 TI - Solution of an insight problem at different age levels. AB - The purpose was to study performance on an insight problem by 3- to 25-yr.-olds. A task involving restructuring and requiring two wooden blocks be fitted together to form a tetrahedron was presented to five groups of 20 subjects each from kindergarten, primary school, secondary school, high school, and a university. The frequencies of solvers within each group increased from the first age group to the third but then remained constant. Solution times and frequencies of solutions attempted were not significantly different among the five groups. Perhaps insight does not follow the same developmental trend as other thinking processes. PMID- 7478881 TI - Digit span and other WISC-R scores in the diagnosis of dyslexia in children. AB - The examination of subtest scores on the Wechsler Intelligence Scale is needed to provide confirmatory evidence for various subtest categorizations as there is no consensus about what patterns might be diagnostically useful. The present study supports the use of the ACID/AVID profiles (Arithmetic, Coding or Vocabulary, Information, and Digit Span) as elements in the diagnosis of dyslexia. WISC-R scores from 44 subjects were analyzed for specific subtest patterns of scores which might separate dyslexic individuals from the WISC-R standardization group. Perhaps some WISC-R ACID/AVID profile pattern analyses may be valid in the diagnosis of a stringently defined population of learning disabled individuals such as dyslexic children. PMID- 7478882 TI - Suicide in Hong Kong, 1971-1990. PMID- 7478883 TI - Comparison of foot and hand reaction times among men: a methodologic study using simple and multiple-choice repeated measurements. AB - The primary study goal was to compare visual simple and choice reaction times of the hand and foot to assess validity of measurements for evaluating subjects' ability to perform rapid, controlled movements. First, we examined the repeatability (N = 34) for four different data-sampling methods from a series of 12 trials within sessions and between two test sessions. Simple and choice reaction times with the preferred hand and both feet were then compared among 153 healthy male volunteers aged 35 to 67 years. Pearson correlations for hand and ipsilateral and contralateral foot reaction times on simple and choice tasks (.53 to .80, p < .001) showed that psychomotor foot reaction time is a valid method for study of neuromuscular control of lower extremities. PMID- 7478884 TI - Handedness and size. PMID- 7478885 TI - Language abilities of neurologically normal and suspect preterm children now in preschool. AB - Multiple aspects of speech and language performance of 29 four- to five-year-old preterm children (10 neurologically suspect) were assessed. Data were collected using a variety of measures including standardized tests and analysis of language samples. Each child's performance was compared with reference data on the various outcomes which yielded an over-all pass score and individual profiles of performance. Neurologically normal children had significantly higher passing scores than suspect children but appeared to be at risk for mild language delays, particularly in morphosyntactic development. Neurologically suspect children appeared to be at risk for moderate and more generalized language delays, including morphosyntactic and phonological development. PMID- 7478886 TI - Relationship between isolated sleep paralysis and geomagnetic influences: a case study. AB - This preliminary report, of a longitudinal study, looks at the relationship between geomagnetic activity and the incidence of isolated sleep paralysis over a 23.5-mo. period. The author, who has frequently and for the last 24 years experienced isolated sleep paralysis was the subject. In addition, incidence of lucid dreaming, vivid dreams, and total dream frequency were looked at with respect to geomagnetic activity. The data were in the form of dream-recall frequency recorded in a diary. These frequency data were correlated with geomagnetic activity k-index values obtained from two observatories. A significant correlation was obtained between periods of local geomagnetic activity and the incidence of isolated sleep paralysis. Specifically, periods of relatively quiet geomagnetic activity were significantly associated with an increased incidence of episodes. PMID- 7478887 TI - Comparison of individual versus group administration of the Modified Version of the Bender-Gestalt Test. PMID- 7478888 TI - Functional brain organization in bipolar affective patients during manic phase and after recovery: a digit dichotic listening study. AB - To study the functional organization of the cerebral hemispheres in patients with bipolar psychosis using a verbal dichotic listening test for pairs of digits 26 patients were tested twice, during the acute expression of manic phase and after recovery. The patient group during the manic phase did not support the expected right-ear advantage of normal subjects on verbal dichotic tests but showed a statistically significant left-ear advantage, which shifted after recovery toward the typical normal asymmetry. Comparing patients during the manic phase and after recovery showed that the left-ear advantage as well as the shift in right-ear advantage after recovery was due to the reduction of left-ear performance. From the over-all neuropsychological findings for these patients mania may be hypothesized to be characterized by overactivation of the right hemisphere. This phaenomenon seems associated with acuteness of the symptoms of the psychotic disorder. PMID- 7478890 TI - Handedness and personality. PMID- 7478889 TI - Self-others perception in a clinical sample of obese women. AB - The purpose of this research, using correspondence analysis on responses to semantic differential scores, was both to examine self-perception in a clinical sample of 120 obese women and to compare this with their attitudes toward obese, thin, and normal persons. Analyses allow us to conclude that, if obese women, as reported previously, consider obesity as a largely negative condition, they have an ambivalent attitude towards themselves. Even though they share the negative connotations socially attributed to obesity, emotionally these obese women not only tend not to recognize their "abnormality" but strongly desire it. PMID- 7478891 TI - Effect of orientation in haptic reproduction of line length. AB - We investigated the accuracy of haptic reproduction of line length and whether accuracy is influenced by line orientation. 13 blindfolded subjects felt along different line lengths at various orientations in the horizontal plane, then reproduced the line lengths in the same orientation as that felt. Efforts were made to equate learning and reproductive scanning movements. Reproductions of line lengths were a nonveridical power function of their true lengths, but the power function exponents did not differ across spatial orientations. It was concluded that people can encode line lengths across spatial orientations by a common power function if care is taken to equate proprioceptive information across learning and reproduction. PMID- 7478892 TI - Interference with olfactory memory by visual and verbal tasks. AB - It has been claimed that olfactory memory is distinct from memory in other modalities. This study investigated the effectiveness of visual and verbal tasks in interfering with olfactory memory and included methodological changes from other recent studies. Subjects were allocated to one of four experimental conditions involving interference tasks [no interference task; visual task; verbal task; visual-plus-verbal task] and presented 15 target odours. Either recognition of the odours or free recall of the odour names was tested on one occasion, either within 15 minutes of presentation or one week later. Recognition and recall performance both showed effects of interference of visual and verbal tasks but there was no effect for time of testing. While the results may be accommodated within a dual coding framework, further work is indicated to resolve theoretical issues relating to task complexity. PMID- 7478893 TI - Analysis by gender and Visual Imagery Reactivity of conventional and imagery Rorschach. AB - Examined here are the effects of gender and Visual Imagery Reactivity in 80 consecutively selected psychiatric outpatients. The participants were grouped by gender and by the amounts of responsiveness to preceding therapy work using imagery (Imagery Nonreactors and Reactors). In the group of Imagery Nonreactors were 13 men and 22 women, and in the Reactor group were 17 men and 28 women. Compared were the responses to standard Rorschach (Conventional condition) with visual associations to memory images of Rorschach inkblots (Imagery condition). Responses were scored using the Visual Imagery Reactivity (VIR) scoring system, a general, test-nonspecific scoring method. Nonparametric statistical analysis showed that critical indicators of Imagery Reactivity encoded as High Affect/Conflict score and its derivatives associated with sexual or bizarre content were not significantly associated with gender; neither was Neutral Content score which categorizes "non-Reactivity." These results support the notion that system's criteria of Visual Imagery Reactivity can be applied equally to both men and women for the classification of Imagery Reactors and Nonreactors. Discussed are also the speculative consequences of extending the tolerance range of significance levels for the interaction between Reactivity and sex above the customary limit of p < .05 in borderline cases. The results of such an analysis may imply a trend towards more rigid defensiveness under Imagery and toward lesser verbal productivity in response to either the Conventional or the Imagery task among women who are Nonreactors. In Reactors, men produced significantly more Sexual Reference scores (in the subcategory not associated with High Affect/Conflict) than women, but this could be attributed to the effect of tester's and subjects' gender combined. PMID- 7478894 TI - Projective personality tests of children with inflammatory bowel disease. AB - 15 children with inflammatory bowel disease were given projective tests after initial diagnosis. Analysis of responses suggested affective constriction, abandonment anxiety, and depression. The children were of good intelligence with good object relationships and psychosexual differentiation for their ages. Severe psychopathology and bizarre slippage were absent. PMID- 7478895 TI - Measuring the typicality of objects included in environmental scenes: a logistic model for atypicality. AB - Some empirical studies have stated that people usually categorize scenes according to the suitability of their elements. This paper proposes a method of measuring the typicality of naturalistic objects contained in environmental scenes. 517 subjects gave a score of suitability for 110 object-scene pairs. We used a logistic model for the measurements which enabled us to obtain two indexes, atypicality and discrimination. Analysis showed that the objects could be arranged on a numerical scale according to their typicality in a scene, and from this we concluded that logistic models are a useful and powerful method of measuring typicality. PMID- 7478896 TI - Relationship between the Luria-Nebraska Neuropsychological Battery and Woodcock Johnson Tests of Achievement-Revised in children with psychiatric impairment. AB - This study was conducted to estimate the relationship between the Luria-Nebraska Neuropsychological Battery and the Woodcock-Johnson Tests of Achievement-Revised at two ages in a child psychiatric sample (N = 113). Analysis showed significant correspondence between scores on the Luria-Nebraska and the Woodcock-Johnson tests at both ages; however, the former tended to show greater impairment than the latter, particularly for the older group. PMID- 7478897 TI - Short-term memory for color names using Stroop material. AB - A modified Stroop arrangement consisting of color names printed in ink of a different color was used in a release from the proactive inhibition (PI) paradigm. In Exp. 1, half of the subjects were informed prior to a trial whether they were to recall the colors or the color names while the remaining half were not informed in advance. After three trials, half of the subjects were shifted to recalling the other material while control subjects continued with the same recall material. Analysis indicated that (1) when subjects were informed in advance, PI release was found in a color name to color shift, but no PI release was found with a shift from color to color name, and (2) when subjects were not informed in advance, no PI release was observed in either shift condition. Exp. 2 replicated the not-informed condition and introduced a longer stimulus-exposure duration for 160 undergraduates. Except for higher recall scores, the same results were obtained. The results were interpreted in terms of differences in the kinds of encoding operations performed on colors and color names. PMID- 7478898 TI - An auditory event related potential evaluation of sonar task experience and age. AB - To gauge the interaction of real-world sonar-task experience and age on brain electrical activity, the effect of sonar experience and age on event related potentials (ERP) was examined. A three-group design was used and the results suggest that sonar experience and age affect the amplitude and distribution of the ERP component. The results concerning age and ERPs support and extend the results of previous studies and suggest that age-related differences occur at a much younger age than is reported elsewhere. Attentional and stimulus evaluation processes which have been linked to parameters of the ERP component may be enhanced with real-world auditory task experience. Research on ERP should control for the possible confounds of auditory-task experience and age. PMID- 7478899 TI - Correlations between the Hand Test Pathology score and Personality Assessment Inventory scales for pain clinic patients. AB - Pearson correlations between the Hand Test Pathology (PATH) score and Personality Assessment Inventory scales produced a cluster of relationships characteristic of an antisocial orientation. Likewise, PATH significantly differentiated between a "P" (Pathology) group flagged by a high Negative Impression score on the inventory, and an "N" (Normal) group of 100 pain patients. It was suggested that the interpretive simplicity of Hand Test scores renders the scores amenable to further correlational studies involving the inventory. PMID- 7478900 TI - Comparison of three psychophysical techniques for estimating body-size perception. AB - Perception of body size was recorded for 63 university students (M age = 25.3 yr., 41 women) who estimated their own body size using three methods. Using the method of adjustment, subjects over- or underestimated their body size. A signal detection analysis indicated that subjects were sensitive to detecting a 4% distortion in body size and that there was no systematic bias for reporting distortion as present or absent. Scores on the adaptive probit estimation task were significantly correlated with values for point of subjective equality and the size judgements with the method of adjustment. Over-all, this experiment demonstrated adaptive probit estimation as a reliable indicator of perceived body size, sensitivity in detecting size distortion, and response bias in making body size judgements. PMID- 7478901 TI - Weber then and now. PMID- 7478902 TI - Edge computation in human vision: anisotropy in the combining of oriented filters. AB - Above threshold, two superimposed sinusoidal gratings of the same spatial frequency (eg 1 cycle deg-1) and equal contrasts, and with orientations balanced around vertical, usually look like a compound structure containing vertical and horizontal edges. However, at large plaid angles (ie large differences between component orientations) and low plaid contrasts there is a tendency for the stimulus to appear as two overlapping gratings (component structure) with obliquely oriented edges. These dependencies of perceived spatial structure in plaids are incompatible with an edge-coding scheme that uses only circular filters to compute zero-crossings, but instead support the idea that different oriented filters can (compound percept) or cannot (component percept) be combined before edges are represented. Here, further evidence is presented in support of this hypothesis. Two-component plaid stimuli had plaid angles of 45 degrees or 90 degrees, and a range of plaid orientations (ie a range of orientations around which the plaid components were balanced). Observers indicated whether each stimulus was perceived as a compound or component structure for a range of plaid contrasts. In addition to angle and contrast effects, perceived spatial structure was also found to depend on plaid orientation: compound structures were perceived more often when the plaid components were balanced around the cardinal axes of the retina. It is suggested that the principles governing the combination of oriented-filter outputs might be learnt during the development of the visual system by using a Hebb-type rule: coactivated filters are more likely to combine their outputs when activated on future occasions. Given the prominence of vertical and horizontal orientations in a carpentered environment, this simple rule promotes a network that combines filters balanced around cardinal axes more readily than oblique axes, in agreement with the results. PMID- 7478903 TI - Respective contribution of orientation contrast and illusion of self-tilt to the rod-and-frame effect. AB - The visual angle subtended by the frame seems to be an important determinant of the contribution of orientation contrast and illusion of self-tilt (ie vection) to the rod-and-frame effect. Indeed, the visuovestibular factor (which produces vection) seems to be predominant in large displays and the contrast effect in small displays. To determine how these two phenomena are combined to account for the rod-and-frame effect, independent estimates of the magnitude of each component in relation to the angular size subtended by the display were examined. Thirty-five observers were exposed to three sets of experimental situations: body adjustment test (illusion of self-tilt only), the tilt illusion (contrast only) and the rod-and-frame test, each display subtending 7, 12, 28, and 45 deg of visual angle. Results showed that errors recorded in the three situations increased linearly with the angular size. Whatever the size of the frame, both mechanisms, contrast effect (tilt illusion) and illusory effect on self orientation (body-adjustment test), are always present. However, rod-and-frame errors became greater at a faster rate than the other two effects as the size of teh stimuli became larger. Neither one nor the other independent phenomenen, nor the combined effect could fully account for the rod-and-frame effect whatever the angular size of the apparatus. PMID- 7478904 TI - The haptic 'oblique effect' in children's and adults' perception of orientation. AB - The haptic perception of vertical, horizontal, and diagonal orientations was studied in children (aged 7 and 9 years) and in adults. The purpose was to test the hypothesis that the haptic oblique effect results from the different scanning movements at work when one hand explores an oblique standard and the other hand sets the response rod. In experiment 1, blindfolded subjects reproduced the orientation of a standard rod presented in either the frontal, the horizontal, or the sagittal plane, and this task was achieved either ipsilaterally (the same hand explored the standard and set the response rod) or contralaterally (one hand explored the standard and the other hand set the response rod). Since, in the sagittal plane, scanning movements are analogous when the left and right hands explore oblique orientations, no oblique effect should be observed in this condition if the hypothesis is valid. Moreover, a development effect should be observed, since young children generally rely more on movement coding than do older children and adults. Results did not support these predictions: the same oblique effect appeared in the frontal and the sagittal planes both in the ipsilateral and in the contralateral condition, and the effect of age was not in the direction predicted by the hypothesis. The results were consistent with the hypothesis in the horizontal plane only. Experiments 2 and 3 provided further tests of this hypothesis but both failed to support it. Taken together, the results of these three experiments did not support the assumption and it is suggested that the haptic oblique effect may be linked to the gravitational cues provided by the arm-hand system when it acts in the three spatial planes. PMID- 7478905 TI - The contribution of stimulus attributes of three-dimensional solid forms and level of discrimination to the visual and haptic percept of balance. AB - Design-trained subjects sorted fourteen solid forms under either visual or haptic conditions into groups on the basis of perceived similarity of balance or along a balance continuum. After these tasks of coarse and fine discrimination, each form was rated on ten bipolar stimulus attributes. Multidimensional scaling and property-fitting analyses were performed on the results of four experiments to determine the combined contribution of stimulus attributes and level of discrimination on the visual and haptic percept of balance. A balance dimension emerged for both modalities, but only when subjects were inclined to attend globally to the structure of a form by the coarse-discrimination task. Results indicate that visual balance is a holistic property of forms which derives from the synthesis of physical stimulus information. For touch, subjects appear to have equated balance with a symmetric distribution of weight/shape about the central axis of a form. Findings are related to theoretical notions of balance. PMID- 7478906 TI - The discrimination of dynamic orientation changes in gratings. AB - Thresholds were measured for discrimination of direction of a step angular rotation of gratings. The addition of simultaneous phase displacements (translation) had little effect on rotation thresholds for gratings over a considerable range; discrimination of rotation is unaffected by random directional translations an order of magnitude larger. Angular rotation discrimination thresholds increased with interstimulus interval (ISI). Thus discrimination is based at short ISIs (180 ms or less) on a percept of rotary motion, but at ISIs of several seconds by a spatial strategy (comparing static component orientations) relying on visual memory. Data points for the short-ISI region fell below the best-fitting straight line, and the slope of the short-ISI region of the curve was steeper than that of the long-ISI region. However, when either compound or simple gratings with uncorrelated spatial frequencies were used in the two stimulus frames, there was no evidence for a separate function at short ISIs. Orientation-change thresholds were measured for simple gratings as a function of contrast and spatial frequency. The contrast function showed saturation and the spatial frequency function was U-shaped. Rotation sensitivity for gratings is thus similar in its spatiotemporal properties to translation sensitivity. The findings support the proposal that rotation discrimination (at short ISIs) is achieved by a template mechanism combining signals from different directional detectors, rather than by congnitive comparison of the outputs of the directional mechanisms themselves. PMID- 7478907 TI - The microgenesis of illusory figures: evidence for visual hypothesis testing. PMID- 7478908 TI - Varying the strength of the Munker-White effect by stereoscopic viewing. AB - In the Munker-White effect grey target bars appear lighter when they are flanked by white bars, and darker when they are flanked by black bars. It is shown that the effect is enhanced if the patterns are presented stereoscopically so that the grey bars appear either behind the grating, in which case they are seen as a rectangle that is occluded by the white bars of the grating, or in front of the grating, so that they form a transparent rectangle. These results are explained in terms of object perception: contrast enhances differences between an object and its surroundings, whereas assimilation reduces differences within an object. PMID- 7478909 TI - 100 years of Benham's top in colour science. AB - For 100 years Benham's top has been a popular device demonstrating pattern induced flicker colours (PIFCs). Results of early and recent investigations on PIFCs are reported and show that the phenomenon originates in phase-sensitive lateral interactions of modulated neural activity in the retina followed by additional spatial interactions in the visual cortex behind the locus of binocular fusion. Colour matches with normal colour stimuli indicate that S/(M + L) opponent neurons are involved. Dichromats do not find matching stimuli for all PIFCs. PIFCs may become useful in medical diagnosis. The phenomenon is interpreted as a side effect of a neural mechanism providing colour constancy under normal stimulus conditions. PMID- 7478910 TI - Reduced sweating threshold during exercise-induced hyperthermia. PMID- 7478911 TI - Na+-HCO3(-) cotransporter and intracellular pH regulation in chicken enterocytes. AB - The current studies examine the presence of the Na+-HCO3(-) cotransporter in chicken enterocytes and its role in cytosolic pH (pHi) regulation. The pH sensitive dye 2',7'-bis(carboxyethyl)-5,6-carboxy-fluorescein (BCECF) was used to monitor pHi. Under resting conditions, pHi was 7.25 in solutions buffered with bis(2-hydroxyethyl)-1-piperazine ethanesulphonic acid (HEPES) and 7.17 in those buffered with HCO3(-). Removal of external Na+ decreased pHi and readdition of Na+ rapidly increased pHi towards the control values. These Na+-dependent changes were greater in HCO3(-)- than in HEPES-buffered solutions. In HCO3- - free solutions the Na+-dependent changes in pHi were prevented by 5-(N-ethyl-N isopropyl)-amiloride (EIPA) and unaffected by 4,4'-diisothiocyanatostilbene disulphonic acid (H2-DIDS). In the presence of HCO3-, the Na+-induced changes in pHi were sensitive to both EIPA and H2-DIDS. In the presence of EIPA, cells partially recovered from a moderate acid load only when both Na+ and HCO3- were present. This pHi recovery, which was EIPA resistant, and dependent on Na+ and HCO3-, was inhibited by H2-DIDS and occurred at equal rates in both Cl- containing and Cl--free solutions. Kinetic analysis of the rate of HCO3- and Na+ dependent pHi recovery from an acid load as a function of the Na+ concentration revealed first-order kinetics with a Michaelis constant, Km, of 11 mmol/l Na+. It is concluded that in HCO3(-) buffered solutions both the Na+/H+ exchanger and the Na+-HCO3(-) cotransporter participate in setting the resting pHi in isolated chicken enterocytes and help the recovery from acid loads. PMID- 7478912 TI - Analysis of evoked and spontaneous quantal release at high pressure in crustacean excitatory synapses. AB - The cellular mechanisms underlying the effect of high pressure on synaptic transmission were studied in the opener muscle of the lobster walking leg. Excitatory postsynaptic currents (EPSCs) were recorded using a loose macropatch clamp technique at normal pressure and 3.5, 6.9 MPa helium pressure. Responses of the single excitatory axon could be grouped into two types: low-yield (L) synapses exhibiting small EPSCs with a considerable number of failures, and high yield (H) synapses having larger EPSCs with very few failures. High pressure reduced the average EPSC amplitude in all synapses and shifted their amplitude histograms to the left by decreasing the quantal content (m) without changing their quantum current (q). A binomial distribution fit of EPSC amplitudes revealed that high pressure greatly decreased n, the number of available active zones, but the effect on p, the probability of release for each zone, was not consistent. Many of the spontaneous miniature EPSCs (mEPSCs), observed only in L type synapses, were "giant" (size = 2-5 q). High pressure increased the frequency of the giant mEPSCs but had little effect on their amplitude histogram. High pressure depressed evoked synaptic transmission by modulating the presynaptic quantal release parameters, but concomitantly enhanced spontaneous quantal release by an unknown mechanism. PMID- 7478913 TI - Cholecystokinin-stimulated enzyme secretion from dispersed rabbit pancreatic acinar cells: phosphorylation-dependent changes in potency and efficacy. AB - In order to establish a regulatory role for phosphoproteins in receptor stimulated enzyme secretion, dispersed rabbit pancreatic acinar cells were stimulated with the COOH-terminal octapeptide of cholecystokinin (CCK8) in the absence and presence of staurosporine and/or 12-O-tetradecanoylphorbol 13-acetate (TPA) or forskolin. The dose/response curve for the stimulatory effect of CCK8 on amylase secretion was biphasic, with a mean half-maximal concentration (EC50) of 21 pM. Staurosporine (1 microM) did not affect secretion elicited by CCK8 concentrations below 0.1 nM, but reduced the response to CCK8 concentrations above 0.1 nM. As a result, the mean EC50 for CCK8 decreased to 8 pM and its efficacy to 70%. The phorbol ester TPA (0.1 microM) attenuated secretion evoked by CCK8 concentrations below 0.1 nM and potentiated the response to CCK8 concentrations above 0.1 nM. As a result, the mean EC50 for CCK8 increased to 0.14 nM and its efficacy to 300%. Staurosporine abolished both the inhibitory and the potentiating effect of TPA, thereby turning the inhibitory effect into a strong potentiating effect. As a result, the mean EC50 for CCK8 decreased to 3 pM, whereas its efficacy increased to 190%. Forskolin (30 microM) potentiated the response to both the lower and the higher CCK8 concentrations. As a result, the mean EC50 for CCK8 increased to 28 pM and its efficacy to 300%. Staurosporine enhanced the potentiating effect of forskolin at CCK8 concentrations below 0.1 nM, but abolished potentiation at CCK8 concentrations above 0.1 nM. As a result, the mean EC50 for CCK8 decreased to 1.4 pM, whereas its efficacy increased to 260%. The data presented demonstrate that the apparent sensitivity of dispersed pancreatic acinar cells to stimulation of the process of enzyme secretion by CCK8 decreases when kinases are activated and increases when kinases are inactivated. Moreover, they show that the efficacy of CCK8 increases by the action of kinases, both sensitive and insensitive to staurosporine. PMID- 7478914 TI - Expression of parathyroid hormone receptors in MDCK and LLC-PK1 cells. AB - Parathyroid hormone (PTH) inhibits renal proximal tubular phosphate (Pi) and bicarbonate reabsorption by regulating the activity of apical Na/Pi cotransport and Na/H exchange. Two renal epithelial cell lines ["proximal tubular", LLC-PK1; "distal tubular", Madin-Darby canine kidney, (MDCK) cells] were stably transfected with complementary deoxyribonucleic acids (cDNAs) encoding a cloned PTH receptor in order to examine the polarity of transfected receptor function and whether or not intrinsic Pi transport is regulated by the transfected PTH receptor. The receptors are functionally coupled to the stimulation of adenosine 3':5' cyclic monophosphate (cAMP) production at both cell surfaces in LLC-PK1 cells, whereas this response is primarily limited to the basolateral surface in MDCK cells. Immunocytochemistry suggests an apical and basolateral localization of the transfected PTH receptor in LLC-PK1 cells and only a basolateral localization in MDCK cells. PTH activation of the transfected receptors is not coupled to the regulation of intrinsic Pi transport in either LLC-PK1 or MDCK cells. PMID- 7478915 TI - Modulation of Ca2+ oscillation and apamin-sensitive, Ca2+-activated K+ current in rat gonadotropes. AB - In rat pituitary gonadotropes, gonadotropin-releasing hormone (GnRH) stimulates rhythmic release of Ca2+ from stores sensitive to inositol 1,4,5-trisphosphate [Ins(1,4,5)P3], which in turn induces an oscillatory activation of apamin sensitive Ca2+-activated K+ current, IK(Ca). Since GnRH also activates protein kinase C (PKC), we investigate the action of PKC while simultaneously measuring intracellular Ca2+ concentration ([Ca2+]i) and IK(Ca). Stimulation of PKC by application of phorbol 12-myristate 13-acetate (PMA) did not affect basal [Ca2+]i. However, PMA or phorbol 12,13-dibutyrate (PdBu), but not the inactive 4alpha-phorbol 12,13-didecanoate (4alpha-PDD), reduced the frequency of GnRH induced [Ca2+]i oscillation and augmented the IK(Ca) induced by any given level of [Ca2+]i. The slowing of oscillations and the enhancement of IK(Ca) were mimicked by synthetic diacylglycerol (1,2-dioctanoyl-sn-glycerol) and could be induced during ongoing oscillations that had been initiated irreversibly in cells loaded with guanosine 5'-O-(3-thiotriphosphate) (GTP-[gammaS]). In contrast, when oscillations were initiated by loading cells with Ins(1,4,5)P3, phorbol esters enhanced IK(Ca) without affecting the frequency of oscillation. The protein kinase inhibitor, staurosporine, reduced IK(Ca) without affecting [Ca2+]i and partially reversed the phorbol-ester-induced slowing of oscillation. Therefore, activation of PKC has two rapid effects on gonadotropes. It slows [Ca2+]i oscillations probably by actions on phospholipase C, and it enhances IK(Ca) probably by a direct action on the channels. PMID- 7478916 TI - Ca2+ influx induced by store release and cytosolic Ca2+ chelation in Ht29 colonic carcinoma cells. AB - Cl- secretion in HT29 cells is regulated by agonists such as carbachol, neurotensin and adenosine 5'-triphosphate (ATP). These agonists induce Ca2+ store release as well as Ca2+ influx from the extracellular space. The increase in cytosolic Ca2+ enhances the Cl- and K+ conductances of these cells. Removal of extracellular Ca2+ strongly attenuates the secretory response to the above mentioned agonists. The present study utilises patch-clamp methods to characterise the Ca2+ influx pathway. Inhibitors which have been shown previously to inhibit non-selective cation channels, such as flufenamate (0.1 mmol.l-1, n = 6) and Gd3+ (10 micromol.l-1, n=6) inhibited ATP (0.1 mmol.l-1) induced increases in whole-cell conductance (Gm). When Cl- and K+ currents were inhibited by the presence of Cs2SO4 in the patch pipette and gluconate in the bath, ATP (0.1 mmol.l-1) still induced a significant increase in Gm from 1.2 +/- 0.3 nS to 4.7 +/- 1 nS (n = 24). This suggests that ATP induces a cation influx with a conductance of approximately 3-4 nS. This cation influx was inhibited by flufenamate (0.1 mmol.l-1, n = 6) and Gd3+ (10 micromol.l-1, n = 9). When Ba2+ (5 mmol.l-1) and 4,4'-diisothiocyanato-stilbene-2-2'-disulphonic acid (DIDS, 0.1 mmol.l-1) were added to the KCl/K-gluconate pipette solution to inhibit K+ and Cl currents and the cells were clamped to depolarised voltages, ATP (0.1 mmol.l-1) reduced the membrane current (Im) significantly from 86 +/- 14 pA to 54 +/- 11 pA (n = 13), unmasking a cation inward current. In another series, the cation inward current was activated by dialysing the cell with a KCl/K-gluconate solution containing 5-10 mmol.l-1 1,2-bis-(2-aminoethoxy)ethane-N,N,N',N'-tetraacetic acid (EGTA) or 1,2-bis-(2-aminophenoxy) ethane-N,N,N',N'-tetraacetic acid (BAPTA). The zero-current membrane voltage (Vm) and Im (at a clamp voltage of +10 mV) were monitored as a function of time. A new steady-state was reached 30-120 s after membrane rupture. Vm depolarised significantly from -33 +/- 2 mV to -12 +/- 1 mV, and Im fell significantly from 17 +/- 2 pA to 8.9 +/- 1.0 pA (n = 71). This negative current, representing a cation inward current, was activated when Ca2+ stores were emptied and was reduced significantly ( Im) when Ca2+ and/or Na+ were removed from the bathing solution: removal of Ca2+ in the absence of Na+ caused a Im of 5.0 +/- 1.2 pA (n = 12); removal of Na+ in the absence of Ca2+ caused a Im of 12.8 +/- 3.5 pA (n = 4). The cation inward current was also reduced significantly by La3+, Gd3+, and flufenamate. We conclude that store depletion induces a Ca2+/Na+ influx current in these cells. With 145 mmol.l-1 Na+ and 1 mmol.l-1 Ca2+, both ions contribute to this cation inward current. This current is an important component in the agonist-regulated secretory response. PMID- 7478918 TI - The selectivity of different external binding sites for quaternary ammonium ions in cloned potassium channels. AB - Tetraethylammonium (TEA) is thought to be the most effective quaternary ammonium (QA) ion blocker at the external site of K+ channels, and small changes to the TEA ion reduce its potency. To examine the properties of the external QA receptor, we applied a variety of QA ions to excised patches from human embryonic kidney cells or Xenopus oocytes transfected with the delayed rectifying K+ channels Kv 2.1 and Kv 3.1. In outside-out patches of Kv 3.1, the relative potencies were TEA > tetrapropylammonium (TPA) > tetrabutylammonium (TBA). In contrast to Kv 3.1, the relative potencies in Kv 2.1 were TBA > TEA > TPA. In Kv 3.1 and Kv 2.1, external tetrapentylammonium (TPeA) blocked K+ currents in a fast, reversible and, in contrast to TEA, time-dependent manner. The external binding of TPeA appeared to be voltage independent, unlike the effects of TPeA applied to inside-out patches. External n-alkyl-triethylammonium compounds (C8, C10 chain length) had a lower affinity than TEA in Kv 3.1, but a higher affinity than TEA in Kv 2.1. In Kv 3.1, the decrease in QA affinity was large when one or two methyl groups were substituted for ethyl groups in TEA, but minor when propyl groups replaced ethyl groups. Changes in the free energy of binding could be correlated to changes in the free energy of hydration of TEA derivatives calculated by continuum methodology. These results reveal a substantial hydrophobic component of external QA ion binding to Kv 2.1, and to a lesser degree to Kv 3.1, in addition to the generally accepted electrostatic interactions. The chain length of hydrophobic TEA derivatives affects the affinity for the hydrophobic binding site, whereas the hydropathy of QA ions determines the electrostatic interaction energy. PMID- 7478917 TI - Lateral hypothalamic lesions facilitate hepatic regeneration after partial hepatectomy in rats. AB - It has been reported that ventromedial hypothalamic lesions facilitate hepatic regeneration through the hepatic vagal nerve after partial hepatectomy. However, whether the lateral area of the hypothalamus is involved in liver regeneration after partial hepatectomy is unknown. To determine the role of the lateral hypothalamic area in this phenomenon, we studied hepatic DNA synthesis during liver regeneration after partial hepatectomy with bilateral lesions of the area. Lesioning of the lateral hypothalamus accelerated the increase in hepatic DNA synthesis and raised the peak level of [methyl-3H]thymidine incorporation after partial hepatectomy. These effects of hypothalamic lesioning were inhibited by combined hepatic vagotomy and sympathectomy. Our results demonstrate that lesioning of the lateral hypothalamus promotes hepatic regeneration through the autonomic nervous system after partial hepatectomy and suggest that the lateral hypothalamic area is involved in liver regeneration through neural mediation. PMID- 7478919 TI - Cardiac channel gating charge movements: recovery from inactivation. AB - The nonlinear charge movements which occur during membrane depolarization of cardiac ventricular myocytes (QON) have been previously identified and separated, by kinetic and steady-state criteria, into constituent components arising from the gating of Na channels and Ca channels. In contrast, the nature and time course of the OFF charge movements (QOFF), which follow membrane repolarization have not been as clearly established. In order to address this question cardiac QOFF was studied using small-diameter, 17-day-old embryonic chick ventricular myocytes that can be rapidly and uniformly voltage-clamped. The application of brief (5.4 ms) depolarizing steps were employed to produce Na channel inactivation but little Ca channel inactivation. Following the return of the membrane potential to -100 mV QOFF, measured as the gating current termed IgOFF, displayed two kinetic components. Double exponential fits to IgOFF yielded time constants of a few tenths of a millisecond for the fast component (IgOFFfast) and of 1-2 ms for the slower component (IgOFFslow). The time course and voltage dependence for the slower component suggested that it might be linked to the inactivation, and the recovery from inactivation, of Na channels. In order to identify these kinetic components double-pulse protocols were employed in which the duration of the prepulse and the interval separating the prepulse and test pulse were varied. The time course for the decay of IgOFFslow following a brief inactivating prepulse was similar to the time course for the recovery of the Na channel QON (QNaRecov). Both IgOFFslow and QNaRecov preceded the recovery of the Na channel (ionic) current. The recovery from inactivation of both the Na current and QNa displayed a similar voltage dependence. These experiments have helped to identify the two components of cardiac IgOFF and, therefore, will facilitate the interpretation of further biophysical and pharmacological studies concerning cardiac Na channel and Ca channel gating charge movements. PMID- 7478920 TI - Effects of inhibitors of enzymatic and cellular pH-regulating systems on central sympathetic chemosensitivity. AB - Previous studies in cats using isolated NaCl-CO2 perfusion of the lower brainstem demonstrated an intrinsic chemosensitivity of sympathoexcitatory bulbospinal neurones within the rostroventrolateral medulla (RVLM). In the present experiments, the effects of inhibitors of enzymatic and cellular systems, known to be involved in pH regulation, were investigated. Isolated perfusion of the lower brainstem with CO2-enriched solutions was performed and preganglionic sympathetic nerve activity (SNA) was recorded. Drugs were locally injected into the left RVLM with glass micropipettes. Perfusion of the RVLM with CO2-enriched solutions over a period of 15 s induced a marked increase in SNA. The magnitude of absolute changes in SNA during perfusion depended on the level of basal SNA before perfusion. Microinjections of 4,4'-diisothiocyanatostilbene-2,2' disulphonic acid (DIDS) and acetazolamide (ACZ) induced a marked rise in basal SNA, whereas diethylpyrocarbonate (DEPC) and ethylisopropylamiloride (EIPA) had no significant effect on basal SNA. After application of DIDS and DEPC, the peak change in SNA due to perfusion of the RVLM with CO2-enriched solutions was slightly diminished. Furthermore, neither ACZ nor EIPA produced any significant influence on the slope, peak change and time course of the increase in SNA compared with control perfusions. We conclude that the enzymatic and cellular carrier systems tested in this study are not or only slightly involved in central sympathetic chemosensitivity. PMID- 7478921 TI - Role of [Ca2+]i in lethal oxidative injury in rat cultured inner medullary collecting duct cells. AB - Reactive oxygen metabolites have been implicated in the pathogenesis of toxic, ischaemic and immunologically mediated renal injury. An increase in the cytosolic free Ca2+ concentration ([Ca2+]i) has been proposed as a mechanism of oxidative stress-induced cell injury. We used a fluorescence spectrometer and a fluorescence probe to measure the [Ca2+]i and viability of rat primary cultured inner medullary collecting duct (IMCD) cells during oxidative stress induced by 5 mM tert-butyl hydroperoxide (TBHP). Initially, this oxidative stress evoked a small increase in [Ca2+]i which was followed by a slower sustained increase from the resting level of 170.8 +/- 38.8 nM to 1490.5 +/- 301.7 nM after 60 min, and this preceded the loss of plasma membrane integrity, measured by the propidium iodide fluorescence method. The elimination of extracellular Ca2+ from the culture medium prevented the TBHP-induced [Ca2+]i increase and improved cell viability. Restoration of extracellular Ca2+ resulted in an immediate and large increase in [Ca2+]i and extensive cell death. Verapamil, a Ca2+ channel blocker, inhibited the [Ca2+]i increase and afforded significant protection against cellular injury following exposure to TBHP-induced oxidative stress. Extracellular acidosis also prevented the increase in [Ca2+]i and cell death caused by this oxidative stress. These results are consistent with the hypothesis that oxidative stress-induced IMCD cellular injury may be the result of increased [Ca2+]i caused, in part, by activation of voltage-dependent Ca2+ channels. PMID- 7478922 TI - Chloride secretion induced by phorbol dibutyrate and forskolin in the human colonic carcinoma cell line HT-29Cl.19A is regulated by different mechanisms. AB - The human colonic carcinoma cell line HT-29cl.19A responds to the protein kinase C activator PDB (4-beta-phorbol 12,13-dibutyrate), as it does to forskolin (an activator of adenylyl cyclase), with a secretory response when the cells are grown on filters and studied at 36 degrees C. Previously, we showed that when cells were grown on Petri dishes and studied at about 25 degrees C with the cell attached patch-clamp technique, forskolin, but not PDB, could activate 8-pS chloride channels (cystic fibrosis transmembrane conductance regulator, CFTR, channels). The present work was carried out to study this discrepancy. Experiments in Ussing chambers, at different temperatures, showed that the responses to PDB and forskolin differ in their temperature sensitivity. This was also found following conventional microelectrode and Ussing chamber studies with nystatin-permeabilized epithelial layers carried out at 25 degrees C and at 36 degrees C. Pre-incubation with the microtubular disruptive agents nocodazole or colcemid did not affect the response to PDB or forskolin, suggesting that chloride secretion induced by these agonists in these cells is independent of the microtubular structure. Pre-incubation with brefeldin A strongly inhibited the response to PDB, but the response to forskolin was hardly affected. The differing effect of temperature and brefeldin A on the responses to forskolin and PDB may be due to the activation of two distinct mechanisms by protein kinases A and C. PMID- 7478923 TI - Effects of diadenosine polyphosphates, ATP and angiotensin II on membrane voltage and membrane conductances of rat mesangial cells. AB - Diadenosine polyphosphates have been shown to influence renal perfusion pressure. As mesangial cells may contribute to these effects we investigated the effects of diadenosine triphosphate (Ap3A), diadenosine tetraphosphate (Ap4A), diadenosine pentaphosphate (Ap5A) and diadenosine hexaphosphate (Ap6A) on membrane voltage (Vm) and membrane conductance (gm) in mesangial cells (MC) of normotensive Wistar Kyoto (WKY) and spontaneously hypertensive (SHR) rats in primary and long-term culture. We applied the patch-clamp technique in the fast-whole-cell configuration to measure Vm and gm. To compare the effects of diadenosine polyphosphates with hitherto known agonists we also tested adenosine 5´ triphosphate (ATP) and angiotensin II (Ang II). As there was no significant difference in the Vm values in MC of WKY (-42 +/- 1 mV, n = 70) and SHR rats (-45 +/- 2 mV, n = 99) as well as in the agonist-induced changes of Vm, all data were pooled. The Vm of all the cells was -44 +/- 1 mV (n = 169) and gm was 15.9 +/- 1.8 nS (n = 141). Ion-exchange experiments showed the presence of a K+ and a non selective cation conductance in resting MC whereas a Cl- conductance or a Na+ selective conductance could not be observed. Ap3A, Ap4A, Ap5A, AP6A and ATP each at a concentration of 5 micromol/l, led to a significant depolarization of Vm by 5 +/- 2 mV (n = 14), 7 +/- 1 mV (n = 25), 3 +/- 1 mV (n = 23), 2 +/- 1 mV (n = 16), and 14 +/- 2 mV (n = 23), respectively. For Ap4A, the most potent diadenosine polyphosphate, we determined the half-maximally effective concentration (EC50) as 6 micromol/l (n = 5-25), for ATP as 2 micromol/l (n = 9 37), and for Ang II as 8 nmol/l (n = 6-18). Ap4A 100 micromol/l increased gm significantly by 55 +/- 20% (n = 16), 100 micromol/l ATP by 135 +/- 60% (n = 18). The diadenosine polyphosphates examined were able to depolarize Vm (Ang II > ATP > Ap4A > Ap3A > Ap5A > Ap6A) by activation of a Cl- conductance and a non selective cation conductance, as do ATP or Ang II. PMID- 7478924 TI - Effects of diadenosine polyphosphates, ATP and angiotensin II on cytosolic Ca2+ activity and contraction of rat mesangial cells. AB - Diadenosine polyphosphates (Apn A) are known to influence cellular Ca2+ activity ([Ca2+]i) in several cells. Their vasoactive potency has been described in various systems including the kidney. We examined the effects of diadenosine polyphosphates, adenosine 5'-triphosphate (ATP) and angiotensin II (Ang II) on cytosolic Ca2+ activity of mesangial cells (MC) in culture obtained from normotensive Wistar-Kyoto (WKY) and spontaneously hypertensive (SHR) rats. [Ca2+]i was measured as a fluorescence ratio F340/F380 with the fura-2 technique using three excitation wavelengths (340 nm, 360 nm and 380 nm) and a photon counting tube. Resting [Ca2+]i was not significantly different in MC from WKY and SHR rats and was measured as 132 +/- 9 nmol/l (n = 65) and 114 +/- 12 nmol/l (n = 36), respectively. Diadenosine polyphosphates (Ap3A-Ap6A) increased [Ca2+]i transiently with an initial peak and a secondary plateau phase comparable to the effects of ATP or Ang II. Increases in [Ca2+]i induced by all these agonists were not significantly different between MC of WKY and SHR rats. ATP, Ap3A, Ap4A, Ap5A, Ap6A (each 5 micromol/l) increased the fura-2 fluorescence ratio initially by 0.66 +/- 0.09 (n = 33), 0.52 +/- 0.08 (n = 18), 0.25 +/- 0.05 (n = 16), 0.09 +/- 0.06 (n = 7), 0.09 +/- 0.04 (n = 11), respectively. A half-maximal initial increase in the fura-2 fluorescence ratio was reached at 22 nmol/l, 0.9 micromol/l, 2.0 micromol/l and 4.0 micromol/l with Ang II, Ap3A, ATP and Ap4A, respectively. Ap4A (100 micromol/l, n = 18) led to a reversible contraction of MC. Diadenosine polyphosphates increase [Ca2+]i in rat MC, in a similar manner to ATP or Ang II and lead to a contraction of MC, suggesting that these nucleotides are also involved in the control of glomerular haemodynamics. PMID- 7478925 TI - Promotion of sleep by heat in young rats. AB - The aim of the experiments was to study the effects of a moderate heat load on sleep in young (26-day-old) rats and to determine whether the sleep-promoting effect of heat results from stimulation of the homeostatic sleep process. The changes in sleep-wake activity, electroencephalogram slow wave activity (SWA) during non-rapid eye movement sleep (NREMS) and cortical temperature (Tcrt) were determined during and after long (24-h) and short (2.5-h) heat loads (elevation of ambient temperature from 26 degrees C to 32 degrees C), and after total sleep deprivation (SD) combined with a short-term heat load. The heat exposures elicited increases in Tcrt and rectal temperature (2 and 1.7 degrees C respectively). The long-term heat load induced persistent, albeit slight enhancements in NREMS. Rapid eye movement sleep (REMS) increased with a 12-h delay during the 24-h heat load. Heat elicited an immediate large increase in SWA. After this initial increase, SWA declined and tended to fall below the baseline level during the last 12 h of the 24-h heat load. SWA and REMS were significantly suppressed after termination of 24-h heat loading. The increased SWA during the short-term heat load was not followed by subsequent alterations in sleep when the ambient temperature had returned to normal. However, after the combination of SD with the short-term heat load the durations of NREMS and SWA were significantly enhanced compared with those found after SD at 26 degrees C. The results are interpreted as suggesting that heat increases NREMS in the young rat by the same mechanism as is involved in the enhancement of NREMS after SD: a stimulation of sleep drive. PMID- 7478926 TI - Release of urodilatin from perfused rat kidney and from cultured neonatal rat kidney cells. AB - Release of rat urodilatin (rURO) from isolated perfused rat kidneys and neonatal rat kidney cells could be demonstrated by a specific competitive radioimmunoassay (rURO-RIA) using [125I]rURO as the competitive antigen and an antiserum against the hypothetical rURO-N-terminus, Ala-Gly-Pro-Arg, as concluded from the amino acid sequence of the rat prohormone CDD/ANP-1-126. This antiserum did not react with synthetic rCDD/ANP-99-126, brain natriuretic peptide (BNP), C-type natriuretic peptide (CNP), or human URO (hURO). rURO could be demonstrated in the urine of the perfused rat kidney after an equilibration period of 20 min. After an initial slight decrease in the second 20 min, rURO production remained at almost the same level during the perfusion time of 100 min. A total of approximately 470 fmol.10 -1.g-1 kidney rURO was produced within 80 min. rURO was also produced by neonatal rat kidney cells kept in serum-free Dulbecco's modified Eagles medium. The production of rURO depended on the cultivation time of the cells. It increased up to 3 days reaching 239 +/- 7.5 fmol.h-1.g-1 protein, afterwards it decreased rapidly. The results obtained indicate that the rat kidney produces a peptide of the type A family of natriuretic peptides, which very likely represents the putative rURO. PMID- 7478927 TI - Patch clamped responses from outer hair cells in the intact adult organ of Corti. AB - Outer hair cells (OHCs) from the mammalian cochlea act as both sensory cells and motor cells. We report here whole-cell tight seal recordings of OHC activity in their natural embedding tissue, the intact organ of Corti, using a temporal bone preparation. The mean cell resting potential, -76 +/- 4 mV (n = 19) and input conductance (10 +/- 3 nS at -70 mV) of third turn hair cells were significantly lower than have been found in isolated cells. Two main K+ currents in the cell were identified. One current, activated positive to -100 mV, was reduced by 5 mM BaCl2. The other current, activated above -40 mV, was reduced by 100 microM 4 aminopyridine (4-AP) and by 30 mM tetraethylammonium (TEA). Both of these currents have been also identified in recordings reported from isolated cells. On stepping to different membrane potentials, cells imaged in the organ of Corti changed length by an amount large enough to cause visible distortions in neighbouring cells. By quantifying such distortions we estimate that the forces generated by OHCs can account for the enhanced response to sound required by the cochlear amplifier. PMID- 7478928 TI - Modulation of cardiac sodium channel isoform by cyclic AMP dependent protein kinase does not depend on phosphorylation of serine 1504 in the cytosolic loop interconnecting transmembrane domains III and IV. AB - Both the neuronal IIA as well as the cardiac SkM2 isoform of the pore forming alpha-subunit of voltage dependent sodium channels are modulated by Protein Kinase A. While alphaIIA becomes attenuated upon PKA stimulation, alphaSkM2 becomes upregulated. PKC dependent phosphorylation of a serine, located in the highly conserved cytoplasmatic region between the third and the fourth transmembraneous domain has been found to be a prerequisite for PKA modulation of the alphaIIA isoform. We used site-directed mutagenesis, expression in Xenopus laevis oocytes and the two-electrode voltage clamp technique to test, whether phosphorylation of the corresponding serine in alphaSkM2 is required for the PKA modulation of also the cardiac isoform. The results clearly indicate that serine 1504 does not play a significant role in the PKA modulation of the cardiac sodium channel isoform, further underlining the differential modulation of the two isoforms by identical signal transduction cascades. PMID- 7478929 TI - Force-inhibiting effect of okadaic acid on skinned rat uterus permeabilized with alpha-toxin. AB - Effects of okadai acid (OA) on contractile force in rat uterine uterine muscles permeabilized with alpha-toxin were examined. (1) Contractile force activated by Ca2+(10(-6.5) M to 10(-4.4) M) was suppressed by relatively low concentrations of OA (30 to 300 nM). The suppressed force was further decreased after washed out of OA. (2)Addition of 10 microM OA enhanced force. Whereas, the increased tension level fell to less than the control level after washed out of OA. (3)Okadaic acid methyl ester (methyl okadaate), an OA derivative without protein phosphatase inhibition, did not affect contraction. These results suggest that the force inhibiting effect of OA is a result of interference with contractile elements through inhibition of protein phosphatases (PPs) activity. PMID- 7478930 TI - Role of calmodulin in the activation of carbachol-activated cationic current in guinea-pig gastric antral myocytes. AB - In mammalian gastrointestinal myocytes, it is known that muscarinic stimulation activates nonselective cation channels through a G-protein and a Ca2+-dependent pathway. We recorded inward cationic currents following application of carbachol (ICCh) to guinea-pig gastric myocytes, which were held at -20 mV using the whole cell patch-clamp method. ICCh was suppressed by nicardipine or removal of Ca2+ from the bath solution. The peak value of inward current induced by repetitive applications of carbachol (CCh) decreased progressively (run-down phenomenon). This run-down was significantly alleviated by the addition of calmodulin to the pipette solution (0.15 mg/ml) or by using the perforated-patch whole-cell voltage clamp technique. Moreover, W-7[N-6(aminohexyl)-5-chloro-1 naphthalenesulphonamide], a calmodulin antagonist, was a reversible inhibitor of ICCh. However, @-7 had only a weak inhibitory effect on the same cationic current which was induced by guanosine 5'-O-(3-thiotriphosphate) (GTP?gammaS] 0.2 mM) in the pipette solution. This GTP[gammaS]-induced cationic current was still markedly suppressed by the Ca2+-free bath solution. W-7 itself had a weak inhibitory effect on voltage-operated Ca2+ channels as well as the effects on ICCh. These data suggest that multiple Ca2+-dependent pathways are involved in the activation of CCh-gated cation channels in guinea-pig antral myocytes and a Ca2+/calmodulin-dependent pathway would be one of them. PMID- 7478931 TI - Adenosine 3':5'-cyclic monophosphate mediates a 5-hydroxytryptamine-induced response in neonatal rat motoneurones. AB - 5-Hydroxytryptamine (5-HT) is present in nerve fibres descending from the brainstem Raphe nuclei to motoneurones and its release is thought to exert excitatory actions. 5-HT, applied from the outside, directly depolarizes spinal and cranial motoneurones in slices. This action of 5-HT is mediated, in part, by an inwardly rectifying cationic current, Ih. In cardiac cells, an equivalent current, if, has been shown to be directly activated by adenosine 3':5'-cyclic monophosphate (cAMP) applied to the inside of the patch membrane. By applying the whole-cell method to thin slices of brainstem and spinal cord, we have shown that intracellularly applied camp and extracellularly applied dibutyryl camp or forskolin mimics the inward current induced by 5-ht. The selective cAMP phosphodiesterase inhibitor, Ro 20-1724, clearly prolonged the 5-HT-induced current. Maximal doses of dibutyryl cAMP or forskolin occluded the 5-HT-induced current. The broad spectrum protein kinase inhibitors 1-(5-isoquinolinesulphonyl) 2-methlypiperazine (H-7) and N-[2-(methylamino)ethyl]-5-isoquinolinesulphonamide (H-8) had no effect on the currents induced by 5-HT and forskolin. From these results, we suggest that activation of 5-HT receptors on the motoneuronal membrane stimulates formation of intracellular cAMP, thereby inducing the inward current, possibly by a direct action on Ih. PMID- 7478932 TI - Effects of cGMP-dependent phosphorylation on rat and human connexin43 gap junction channels. AB - The effects of 8-bromoguanosine 3':5'-cyclic monophosphate (8Br-cGMP), a membrane permeant activator of protein kinase G (PKG), were studied on rat and human connexin43 (Cx43), the most abundant gap junction protein in mammalian heart, which were exogenously expressed in SKHep1 cells. Under dual whole-cell voltage clamp conditions, 8Br-cGMP decreased gap junctional conductance (gj) in rat Cx43 transfected cells by 24.0 +/- 3.7% (mean +/- SEM, n = 5), whereas gj was not affected in human Cx43-transfected cells by the same treatment. The relaxation of gj in response to steps in transjunctional voltage observed in rat Cx43 transfectants was best fitted with three exponentials. Time constants and amplitudes of the decay phases changed in the presence of 8Br-cGMP. Single rat and human Cx43 gap junction channels were resolved in the presence of halothane. Under control conditions, three single-channel conductance states (gammaj) of about 20, 40-45 and 70 pS were detected, the events of the intermediate size being most frequently observed. In the presence of 8Br-cGMP, the gammaj distribution shifted to the lower size in rat Cx43 but not in human Cx43 transfectants. Immunoblot analyses of Cx43 in subconfluent cultures of rat Cx43 or human Cx43 transfectants showed that 8Br-cGMP did not induce changes in the electrophoretic mobility of Cx43 in either species. However, the basal incorporation of [32P] into rat Cx43 was significantly altered by 8Br-cGMP, whereas this incorporation of [32P] into human Cx43 was not affected. We conclude that 8Br-cGMP modulates phosphorylation of rat Cx43 in SKHep1 cells, but not of human Cx43. This cGMP-dependent phosphorylation of rat Cx43 is associated with a decreased gj, which results from both an increase in the relative frequency of the lowest conductance state and a change in the kinetics of these channels. PMID- 7478933 TI - The temporal relationship between intraocular pressure and extraocular muscle activation in cats. AB - The temporal relationship between intraocular pressure and extraocular muscle activation was studied in cats in response to the administration of the depolarizing muscle relaxant, succinylcholine (i.e. bolus doses of 0.1 and 1.0 mg/kg). Simultaneous changes in intraocular pressure, extraocular muscle force, extraocular electromyograms (EMGs), limb muscle EMGs and hindlimb muscle afferent activity were recorded. Increases in intraocular pressure were associated with extraocular muscle activation and had two components: (1) an initial abrupt increase (lasting seconds) which correlated with fasciculations within the extraocular and hindlimb muscles; and (2) a latter more sustained component (minutes) presumably due to tonic muscle activation which correlated with increases in hindlimb muscle afferent activity (e.g. due to sustained activation of bag 1 intrafusal fibers by succinylcholine). In a separate group of animals, in which the extraocular muscles were detached from the right eye bilateral intraocular pressures were measured: depolarization by succinylcholine caused a significant increase in intraocular pressure only for the eye with intact muscles. Thus, increases in intraocular pressure following the administration of succinylcholine are directly related to the changes in extraocular muscle tension which is dependent on both tonic and phasic muscle fiber responses. PMID- 7478934 TI - The desensitization of the embryonic mouse muscle acetylcholine receptor depends on the cellular environment. AB - The rate of desensitization of nicotinic acetylcholine (ACh) receptor (nAChR), an important characteristic of nAChR function, was studied in myotubes of the mouse C2C12 cell line at different times after fusion, by measuring the decay of ACh evoked currents (IACh) under various patch-clamp configurations. We observed a progressive slowing of IACh decay rate (half-decay time rose from about 0.5 s to over 5 s) in myotubes of increasing size (i.e. age) under all experimental conditions, except in outside-out patches, when IACh decayed as fast as in the smallest myotubes. Single-channel conductance (about 35 pS) and open time (about 3.5 ms), measured in outside-out and cell-attached patches, were independent of myotube size. In Xenopus oocytes injected with poly(A+)RNA extracted from C2C12 myoblasts or mature myotubes, IACh decay was about 50 times slower than in myotubes. Neither cAMP-dependent nor diacylglycerol-dependent protein kinases, actin nor microtubule polymerization state influenced IACh decay. Our data indicate that the cellular environment, but not readily dialysable cytosolic factors, markedly influences the functional behaviour of nAChR. PMID- 7478935 TI - Immunolocalization of lamins and nuclear pore complex proteins by atomic force microscopy. AB - The nuclear envelope functions as a selective barrier separating the nuclear from the cytosolic compartment. Nuclear pore complexes (NPCs) mediate nuclear import and export of macromolecules and, therefore, are potential regulators of gene expression. In this study we applied atomic force microscopy (AFM) to visualize the three dimensional (3D) structure of individual NPCs in the absence and presence of two different antibodies, one directed against a pore protein (gp62) and another directed against Xenopus lamin LIII, a component of the nuclear lamina, a filament meshwork localized on the nucleoplasmic side of the nuclear envelope (NE) adjacent to and interacting with NPCs. Using 12-nm gold-labelled secondary antibodies and transmission electron microscopy we could clearly localize the primary single anti-gp62 antibody on NPCs and the primary single anti-LIII antibody between NPCs. Using AFM, the secondary antibodies against anti gp62 could be detected as particles 7 nm in height on the nucleoplasmic face of NPCs. The secondary antibodies against anti-LIII could be clearly identified between NPCs. The secondary antibodies, attached to a 12-nm colloidal gold particle and visualized on glass, revealed similar shapes and heights as found on NEs. According to the 3D images, the volume of a single gold particle conjugated with secondary antibodies was 10203 nm3. This volume is equivalent to the volume of 38 IgG molecules associated with one individual gold particle. A similar volume of 11987 nm3 was calculated from a model assuming that the 150-kDa IgG molecules perfectly cover the spherical gold particle. We conclude that AFM can be used for identifying antibodies or other macromolecules associated with biomembranes. PMID- 7478936 TI - Polarized ion transport during migration of transformed Madin-Darby canine kidney cells. AB - Epithelial cells lose their usual polarization during carcinogenesis. Although most malignant tumours are of epithelial origin little is known about ion channels in carcinoma cells. Previously, we observed that migration of transformed Madin-Darby canine kidney (MDCK-F) cells depended on oscillating K+ channel activity. In the present study we examined whether periodic K+ channel activity may cause changes of cell volume, and whether K+ channel activity is distributed in a uniform way in MDCK-F cells. After determining the average volume of MDCK-F cells (2013+/-270 microm3; n=8) by means of atomic force microscopy we deduced volume changes by calculating the K+ efflux during bursts of K+ channel activity. Therefore, we measured the membrane conductance of MDCK-F cells which periodically rose by 22.3+/-2.5 nS from a resting level of 6.5+/-1.4 nS (n=12), and we measured the membrane potential which hyperpolarized in parallel from -35.4+/-1.2 mV to -71.6+/-1.8 mV (n=11). The distribution of K+ channel activity was assessed by locally superfusing the front or rear end of migrating MDCK-F cells with the K+ channel blocker charybdotoxin (CTX). Only exposure of the rear end to CTX inhibited migration providing evidence for "horizontal" polarization of K+ channel activity in transformed MDCK-F cells. This is in contrast to the "vertical" polarization in parent MDCK cells. We propose that the asymmetrical distribution of K+ channel activity is a prerequisite for migration of MDCK-F cells. PMID- 7478937 TI - A fluorimetric and amperometric study of calcium and secretion in isolated mouse pancreatic beta-cells. AB - We have examined the temporal relationship between intracellular Ca2+ concentration ([Ca2+]i) and secretion in single intact pancreatic beta-cells. Secretion was detected as the release of 5-hydroxytryptamine from pre-loaded beta cells, using amperometry, and changes in [Ca2+]i were monitored by microfluorimetry. Stimulation of beta-cells by elevation of the extracellular K+ concentration ([K+]o), acetylcholine or glucose increased [Ca2+]i and, after a delay of 2-7 s, evoked amperometric currents. In the presence of glucose, we observed oscillations in [Ca2+]i which were associated with oscillations in the amplitude and frequency of amperometric currents: however, the temporal correlation was not exact, suggesting that there is a significant latency between the increase in average [Ca2+]i and exocytosis. Both the amplitude and frequency of the amperometric currents elicited by 50 mM KCl declined with successive stimulation, but were restored by agents which elevate intracellular adenosine 3':5':cyclic monophosphate (cAMP). This suggests that beta-cells may possess a readily releasable pool of granules which is replenished by cAMP. The variable amplitude of the amperometric currents is discussed in terms of a model in which several secretory granules fuse simultaneously with the plasma membrane. PMID- 7478938 TI - Protein kinase C consensus sites and the regulation of renal Na/Pi-cotransport (NaPi-2) expressed in XENOPUS laevis oocytes. AB - Renal brush border membrane sodium/phosphate (Na/Pi)-cotransport activity is inhibited by hormonal mechanisms involving activation of protein kinases A and C. The recently cloned rat renal Na/Pi-cotransporter (NaPi-2) contains several protein kinase C but no protein kinase A consensus sites [17, 20]. In the present study we have expressed wild type and polymutant (protein kinase C consensus sites removed) NaPi-2-transporters in Xenopus laevis oocytes. The expression of transport function as well as the basic transport properties were unaffected by the removal of the consensus sites. Pharmacological activation of protein kinase C with phorbol 12,13-didecanoate (PDD) led to a time-dependent inhibition of expressed wild type Na/Pi-cotransport function; simultaneous exposure to staurosporine (0.3) prevented the PDD induced (50 nM) inhibition. The kinase-C mediated inhibition was not prevented by the removal of the protein kinase C consensus sites. Pharmacological activation of protein kinase A (dibutyryl adenosine 3':5':cyclic monophosphate (cAMP)/forskolin) had no effect on wild type NaPi-2-induced oocyte Na/Pi-cotransport. It is concluded that the protein-kinase C-mediated regulation of expressed Na/Pi-cotransport does not involve the predicted consensus sites. The involvement of "cryptic" phosphorylation sites and/or of a phosphorylated "regulatory" protein is discussed. PMID- 7478939 TI - Substrate-charge dependence of stoichiometry shows membrane potential is the driving force for proton-peptide cotransport in rat renal cortex. AB - The proton dependence of the transport of three labelled, hydrolysis-resistant synthetic dipeptides carrying a net charge of -1, 0 or +1 has been investigated in a brush border membrane vesicle preparation obtained from rat renal cortex. Cross-inhibition studies are consistent with the transport of all peptides studied being through a single system. The extent and time course of uptake in response to an inwardly directed electrochemical gradient of protons differed for each peptide. For the cationic peptide D-Phe-L-Lys this gradient did not stimulate the initial rate of uptake, while for the neutral dipeptide D-Phe-L-Ala and the anionic peptide D-Phe-L-Glu stimulation was observed. However, the effect on D-Phe-L-Glu was more marked than that on D-Phe-L-Ala and the proton activation differed for these two peptides. The calculated Hill coefficients for the two proton-dependent peptides were 1.14 +/- 0.16 and 2.15 +/- 0.10 for D-Phe-L-Ala and D-Phe-L-Glu, respectively, providing evidence that the stoichiometry of proton:peptide cotransport is different for each peptide (0:1, 1:1 and 2:1 for D Phe-L-Lys, D-Phe-L-Ala and D-Phe-L-Glu respectively); studies on energetics are compatible with this conclusion. The physiological and molecular implications of this model are discussed, as are the applicability of the conclusions to secondary active transport systems more generally. PMID- 7478940 TI - Transport characteristics of a murine renal Na/Pi-cotransporter. AB - A complementary deoxyribonucleic acid (cDNA) corresponding to a murine renal cortical Na/phosphate-(Na/Pi-) cotransporter was isolated and its transport properties characterized by electrophysiological techniques after expression in Xenopus laevis oocytes. A Na-dependent inward movement of positive charges ("short-circuit current") was observed upon superfusion with Pi (and with arsenate). Increasing the Na concentration led to a sigmoidal elevation in Pi induced short-circuit current; the apparent Michaelis constant, Km, (around 40 mM Na) was increased by lowering the pH of the superfusate but was not influenced by altering the Pi concentration. Increasing the Pi (and arsenate) concentration led to a hyperbolic elevation in Na-dependent short-circuit current (apparent Km for Pi at 100 mM Na was around 0.1 mM; apparent Km for arsenate was around 1 mM); lowering the Na concentration decreased the apparent affinity for Pi. The Pi induced short-circuit current was lower at more acidic pH values (at pH 6.3 it was about 50% of the value at pH 7.8); this pH dependence was similar if the Pi concentration was calculated in total, or if distinction was made between its mono- and divalent forms. Thus, the pH dependence of Na-dependent Pi transport (total Pi) may not be related primarily to a pH-dependent alteration in the availability of divalent Pi, but includes also a competitive interaction of Na with protons. The effect of Pi and Na concentration on the apparent Km values for Na or Pi, respectively, provides evidence for an ordered interaction of "cosubstrate" (Na first) and "substrate" (Pi or arsenate second). PMID- 7478941 TI - Properties of calcium currents and contraction in cultured rat diaphragm muscle. AB - The characterization of calcium currents and contraction simultaneously measured in cultured rat diaphragm muscle cells was carried out in the present study. Whole-cell patch-clamp experiments were designed to further elucidate the mechanism of excitation-contraction (E-C) coupling in diaphragm which, though generally considered a skeletal-type muscle, has been reported to exhibit properties indicative of a cardiac-like E-C coupling mechanism. Normalized current/voltage (I/V) curves for two concentrations of external calcium (2.5 and 5 mM) were obtained from diaphragm myoballs. Both curves showed peaks corresponding to the activation of a T-type calcium current and a dihydropyridine sensitive L-type calcium current. The normalized curve for the voltage dependence of the activation of contraction in diaphragm myoballs followed a typical Boltzmann-type relationship to the peak of contraction. Thereafter, the curve declined in a manner that was more pronounced in diaphragm compared to that measured in additional experiments using cultured rat limb muscle myoballs. This effect could be interpreted in terms of a more pronounced participation of the L type current in E-C coupling in cultured diaphragm muscle. An increased likelihood of cultured diaphragm muscle to undergo depletion of sarcoplasmic reticular calcium stores during repetitive stimulation, or a heightened propensity for the voltage sensor for E-C coupling in diaphragm to enter the inactive state could also explain this effect. Maximal contractile activity was only slightly affected when the L-type current was blocked by externally applied cadmium (2 mM) or cobalt (3 mM), suggesting that a pronounced calcium-current dependent component of contraction is unlikely in cultured diaphragm muscle. These results show that T- and L-type calcium channels are expressed in cultured rat diaphragm muscle cells and that, in contrast to cardiac muscle, the entry of calcium ions via L-type voltage-dependent calcium channels is not a prerequisite for contraction. Differences in the voltage sensitivity of contraction, observed at depolarized membrane potentials in cultured rat diaphragm and limb muscle cells, suggest that the voltage sensor for E-C coupling in diaphragm might more readily enter an inactivated configuration - possibly by a mechanism which is dependent on the concentration of external calcium. PMID- 7478942 TI - Regulation of murine acid secretion by CO2. AB - To determine whether endogenous metabolic sources alone provide sufficient CO2 for acid secretion in mammals, basal and stimulated acid secretion and metabolic CO2 production were measured concurrently in mouse stomachs, in vitro, without exogenous CO2, and after addition of 5% CO2 serosally. Basal acid secretion was varied by changing luminal pH from 3.2 to 4.0. In the absence of an exogenous supply of CO2 acid secretion was stable under basal conditions and increased during cholinergic stimulation with carbachol. Serosal CO2 supply increased basal and stimulated acid secretion. The increase in basal acid secretion depended on the initial level of acid secretion. At pH 4.0, exogenous CO2 increased acid output (mean +/- SD) by 13% from 112 +/- 11 nmol/min to 126 +/- 8 nmol/min (P < 0.03), whereas at pH 3.6 the increase was 40% (63 +/- 14 to 88 +/- 20 nmol/min, P < 0.04) and 157% at pH 3.2 (21 +/- 13 to 54 +/- 14 nmol/min, P < 0.002). Following cholinergic stimulation a maximal acid output of 321 +/- 38 nmol/min was attained without serosal CO2, whilst addition of 5% CO2 to the serosal solution increased maximal acid secretion by 49% to 479 +/- 96 nmol/min (P < 0.005). Metabolic activity, measured as total gastric CO2 production, was greater as acid secretion rates increased [239 +/- 20 nmol/min at 21 +/- 13 nmol/min (luminal pH 3.2) versus 406 +/- 28 nmol/min at 321 +/- 17 nmol/min (after cholinergic stimulation)]. The data support the concept that basal and sub maximal acid secretion can be maintained by CO2 available from metabolic sources, but full expression of the acid secretory apparatus requires exogenous CO2. PMID- 7478943 TI - Red blood cell flow cessation and diameter reductions in skeletal muscle capillaries in vivo - the role of oxygen. AB - When perfusion pressure is reduced, red blood cell flow in the capillaries of skeletal muscle ceases at a positive pressure difference across the vascular bed, while arterioles dilate and venules are not constricted. This flow cessation (i.e., cessation of red blood cell flow) and luminal diameter changes in capillaries following femoral arterial pressure reduction were investigated in the rabbit tenuissimus muscle in situ (n = 42) using intravital video microscopy. Arterial pressure was reduced by occlusion of the aorta distal to the renal arteries. During the experiments, leg and muscle were placed in a sealed box. The muscle was exposed to low PO2 by leading a gas mixture deprived of O2 through the box. Locally at the muscle surface, i.e., under the microscope objective, PO2 was varied by varying the PO2 in the superfusion solution. In all experiments, the remainder of the muscle was kept at low (< 20 mm Hg) PO2. The incidence of flow cessation was virtually zero at low local (< 20 mm Hg) PO2 and became almost 100% at local values above 70 mm Hg. Initial equivalent capillary diameters were 3.1 5.8 microm (median 4.0 microm) and did not correlate with local O2 tension. During aorta occlusion, capillary diameters significantly (P < 0.0001) decreased by a median value of 8% at all local PO2 values; in 14 out of 54 capillaries local diameter became less than 2.8 microm. The extent of diameter reduction did not correlate with PO2. In the 14 capillaries in which the diameter became less than 2.8 microm flow cessation occurred in only four cases. The minimal diameter reached was always at the site of an endothelial nucleus. The capillary diameter reductions are probably due to passive recoil. In the 48 capillaries in which flow ceased, only in four cases did a red blood cell stop at the site of the nucleus. We conclude that capillary diameter reductions (local and generalized) lead to a considerable increase in capillary resistance which contributes to the occurrence of flow cessation but cannot solely explain it. PMID- 7478946 TI - [CT findings of mass-forming pancreatitis: correlation with histopathologic findings]. AB - We attempted to analyse the computed tomographic (CT) characteristics of mass forming pancreatitis. Thirty inflammatory masses were studied using the combined method of dynamic CT (Dy-CT) and high dose enhancement CT (HD-CT) with a thin section scan. From the correlation between the CT enhancement effect and the histopathologic changes, injury of the glandular element and proliferation of fibrous tissue were more prominent in inflammatory masses showing a different enhancement effect from the surrounding pancreatic parenchyma. In addition to these changes, constriction of the intrapancreatic artery caused a poor enhancement effect on Dy-CT, whereas marked round cell infiltration with lymphoid follicle formation increased the enhancement effect on HD-CT. This combined CT method is valuable for the diagnosis of mass-forming pancreatitis and adequately reflects its histopathologic condition. PMID- 7478944 TI - Intrapulmonary gas mixing and dead space in artificially ventilated dogs. AB - In this study we have investigated the effects of breath holding and of the physical properties of gases on four different respiratory dead spaces (VD): the Fowler, the physiological, the washout and the inert gas dead space. The experiments were performed with dogs which were ventilated artificially with breathing patterns with different post-inspiratory breath holding times (ta) of 0, 0.5, 1.0 and 2.0 s. Tracer amounts of acetone, ether and enflurane were infused continuously into a peripheral vein and a bolus of a mixture of krypton, Freon12 and SF6 was introduced into the peritoneal cavity. After reaching steady state, samples of arterial blood, mixed venous blood and mixed expired air were taken simultaneously. From the partial pressures (Pa, PV and PE respectively) we determined the excretion (E = PE/PV), retention (R = Pa/PV) and the physiological dead space fraction (VD,phys/VT = (1- PE/Pa)) for each gas, where VT is tidal volume. Further, we recorded the expirograms of the six tracer gases and of CO2 from which the Fowler dead space fractions (VD,Fowler/VT) of the different gases were determined. Also the washout dead space fractions (VD,washout/VT) for He and SF6 were determined as well as the inert gas dead space fraction (VD,MIGET/VT) with the use of the multiple inert gas elimination technique (MIGET). With the exception of VD,phys/VT for SF6, all dead space fractions decreased with increasing ta. VD,phys/VT for the poorly soluble gas SF6 was considerably larger than VD,phys/VT for the remaining gases. For the highly soluble acetone VFowler/VT was considerably smaller than VD,Fowler/VT for the other gases. VD,washout,SF6/VT was always larger than VD,washout,He/VT and VD,Fowler,SF6/VT. Further, VD,phys/VT was larger than VD,Fowler/VT for SF6 and acetone. However, for gases with intermediate solubility in blood VD,phys/VT tended to be smaller than VD,Fowler/VT. We conclude that the respiratory dead spaces are affected by the breathing pattern and by the physical properties of gases, i.e. their diffusivity in alveolar gas and their solubility in blood or lung tissue. PMID- 7478947 TI - [Experimental study on early detection of alloxan-induced pulmonary injury by magnetic resonance imaging]. AB - We studied the early detection of alloxan-induced pulmonary injury by magnetic resonance imaging in vivo. Permeability edema was induced in ten rats by intravenous injection of alloxan at 100 mg/kg. T1-and T2-weighted images were acquired in five rats every 30 min for 120 min after alloxan injection. Five rats served as controls. The rats were sacrificed immediately after imaging and examined microscopically. CT images were also acquired in five rats every 30 min for 120 min after alloxan injection. Five rats served as controls. The rats were sacrificed immediately after imaging, and the wet-to-dry ratio of the lung was measured. In T1-weighted images, relative signal intensity from the lung with permeability edema rose from 30 min to 120 min, and was greater than that from normal lung every time. In T2-weighted images, there was no statistically significant difference in relative signal intensity of the lung between permeability edema and the control during 120 min. In CT images, there was also no statistically significant difference in lung density between permeability edema and the control during 120 min. There was no statistically significant difference in the wet-to-dry lung ratio between edematous lung and normal lung. In histological study, mild congestion and interstitial edema were observed in edematous lung. These results suggest the potential capability of MR imaging in detecting the early phase of permeability pulmonary edema. PMID- 7478945 TI - [Ca2+]i-dependent membrane currents in guinea-pig ventricular cells in the absence of Na/Ca exchange. AB - Transient inward currents (Iti) during oscillations of intracellular [Ca2+] ([Ca2+]i) in ventricular myocytes have been ascribed to Na/Ca exchange. We have investigated whether other Ca2+-dependent membrane currents contribute to Iti in single guinea-pig ventricular myocytes, by examining membrane currents during [Ca2+]i oscillations and during caffeine-induced Ca2+ release from the sarcoplasmic reticulum in the absence of Na+. Membrane currents were recorded during whole-cell voltage clamp and [Ca2+]i measured simultaneously with fura-2. In the absence of Na/Ca exchange, i.e., with Li+, Cs+ or N-methyl-D-glucamine (NMDG+) substituted for Na+, the cell could be loaded with Ca2+ by repetitive depolarizations to +10 mV, resulting in spontaneous [Ca2+]i oscillations. During these oscillations, no inward currents were seen, but instead spontaneous Ca2+ release was accompanied by a shift of the membrane current in the outward direction at potentials between -40 mV and +60 mV. This [Ca2+]i-dependent outward current shift was not abolished when NMDG+ was substituted for internal monovalent cations, nor was it sensitive to substitution of external Cl-. It was however, sensitive to the blockade of ICa by verapamil. These results suggest that the transient outward current shift observed during spontaneous Ca2+ release represents [Ca2+]i-dependent transient inhibition of ICa. Similarly, during the [Ca2+]i transients induced by brief caffeine (10 mM) applications, we could not detect membrane currents attributable to a Ca2+-activated nonselective cation channel, or to a Ca2+-activated Cl- channel; however, transient Ca2+-dependent inhibition of ICa was again observed. We conclude that neither the Ca2+-activated nonselective cation channel nor the Ca2+-activated Cl- channel contribute significantly to the membrane currents during spontaneous [Ca2+]i oscillations in guinea-pig ventricular myocytes. However, in the voltage range between -40 mV and +60 mV Ca2+-dependent transient inhibition of ICa will contribute to the oscillations of the membrane current. PMID- 7478948 TI - [Evaluation of calcification in pulmonary nodules using bone images obtained with single exposure energy subtraction]. AB - Bone images obtained by single exposure dual energy subtraction with computed radiography, a new image processing method, were evaluated for the detectability of calcification in lung nodules in both phantom and clinical studies. In the phantom study, five simulated nodules containing known and uniformly distributed amounts of calcium hydroxide were placed in a human chest phantom and images were obtained the following five techniques: 1) conventional radiography (CXR), 2) low kV conventional radiography (Low), 3) Fuji computed radiography (FCR), 4) bone images with conventional singles exposure energy subtraction (ES) and 5) bone image with fine single exposure energy subtraction (FES). The detectability of calcification in the simulated nodules was evaluated by six radiologists using ROC analysis and two-way analysis of variance. In the clinical study, images of 42 lung nodules (21 calcified lung nodules and 21 non-calcified nodules), in which calcification was diagnosed by computed tomography, were obtained with the following four techniques: 1) CXR, 2) FCR, 3) ES and 4) FES and the detectability of calcification in the lung nodules was evaluated by nine radiologists using ROC analysis. In the phantom study, FES was significantly superior to CXR, Low and ES, and equal to FCR in the detectability of calcification in the simulated nodules. Compared with ES, FES was especially superior in the underexposed portion of the chest radiograph. In terms of calcium content in the simulated nodules, FES and FCR were superior to CXR in detecting calcification when a lower calcium hydroxide content was present. In the clinical study, FES was significantly superior to CXR, FCR and ES in the diagnosis of calcification in lung nodules. These findings indicate the usefulness of FES in the detection of calcification in lung nodules, and suggests that it is promising for the diagnostic screening of lung nodules. PMID- 7478949 TI - [Interuncal distance measurements in normal controls and patients with dementia: MR imaging study]. AB - To evaluate the utility of measuring interuncal distance (IUD) as a reflection of the limbic system, we compared the IUD of 60 dementia patients with that of 10 normal controls. We also measured the width of the intracranial compartment (W1 and W2) to correct for differences in individual brain size, and calculated the ratio of IUD/W1 and IUD/W2. IUD could not separate patients with dementia from normal controls, but there were significant differences in IUD/W1 and IUD/W2 between patients with dementia and normal controls. IUD, IUD/W1 and IUD/W2 did not correlate with Mini-Mental Examination score or ADAS score in patients with dementia. We conclude that IUD measurement is not helpful in distinguishing patients with mild stage dementia from normal aged people or as a scale for dementia. However, we suggest that IUD/W1 and IUD/W2 can discriminate between cases of mild dementia and normal aged people. PMID- 7478951 TI - [Intra-bile duct growth of hepatocellular carcinoma: value of biliary dilatation on CT]. AB - In order to evaluate the usefulness of CT in demonstrating biliary invasion by hepatocellular carcinoma, 191 surgically proved cases were studied. Among 191 CT scans performed before surgery, six (3%) showed biliary dilatation. Pathological biliary invasion was found in eight cases (4%). Of these eight cases, four cases (50%) showed biliary dilatation on CT. In six cases with biliary dilatation on CT, pathological biliary invasion was revealed in four cases (67%). In two cases, the large (> or = 6cm) encapsulated tumors located in the hepatic hilum dilated the intrahepatic bile duct without intraductal tumor growth. We concluded that biliary dilatation on CT cannot be a sign of biliary invasion by hepatocellular carcinoma. PMID- 7478950 TI - [Computed radiography in X-ray examination of stomach: evaluation by phantom experiments and clinical studies]. AB - The usefulness and clinical applicability of computed radiography (FCR) in X-ray examination of the stomach were evaluated in comparison with film-screen (FS) radiography by both phantom experiments and clinical studies. Receiver operating characteristic (ROC) curve analysis consisting of 25 normal and 25 artificially created simulated lesions on an originally created Styrofoam phantom showed no difference in lesion detectability between FCR and FS radiography. Both were also equal in evaluation of the internal structure and margin of the 25 lesions by visual ranking. ROC curve analysis of clinical cases was performed in 30 cases, 20 normal and 10 abnormal, while evaluation by visual ranking was performed on 18 lesions. There was no significant difference between FCR and FS radiography either with ROC curve analysis or visual ranking, although the variations were somewhat greater in the clinical cases than in the phantom studies. I conclude that FCR is equal to FS radiography in X-ray examination of the stomach regarding both the detectability and visual conspicuity of the lesion. In view of the other advantages of FCR, it can safely replace conventional FS radiography in X-ray examination of the stomach. PMID- 7478952 TI - [Experimental and clinical studies of degradable starch microspheres in the treatment of hepatic neoplasm: Part 1. Experimental study]. AB - An Experimental study was performed to evaluate arterial blood flow changes caused with the intra-arterial administration of degradable starch microspheres (DSM). Hemodynamic and histological changes caused by the intra-arterial administration of DSM were investigated in rabbit kidneys. The degree of blockade of arterial blood flow was correlated with the dose of DSM, and angiography reflected these changes reliably. Histological changes were also dependent on the dose of DSM. Less than 6mg administration had caused little or no histological change. However massive necrosis of the renal tubules had been found with more than 15mg of DSM administration. The degree of blockade of arterial blood flow and histological change could be controlled by changes in the dose of DSM. PMID- 7478953 TI - [Clinical application of magnetic resonance angiography for coronary arteries: correlation with conventional angiography and evaluation of imaging time]. AB - Magnetic resonance angiography (MRA) of the coronary arteries is a particularly difficult task because of the small size of the vessels and cardio-respiratory motion. The authors describe a method of performing of ultrafast MRA of the coronary arteries with a standard MR system and body coil. Each image was obtained within a single breath hold by "segmented Turbo FLASH" sequences using an electrocardiography gate. Clinical application was performed in 20 patients with ischemic heart disease, and a comparison was made with conventional coronary angiography. The imaging time was shortened significantly by our methods. The diagnosis was made by at least two different images to avoid false positives. The results indicated a good correlation between MRA and conventional angiography. PMID- 7478955 TI - [Evaluation of pancreatic cancers using thallium-201 single photon emission computed tomography]. AB - Radionuclide study has not been frequently applied to pancreatic cancers because of the absence of suitable radiopharmaceuticals for their positive depiction. We evaluated thallium-201 chloride (201T1) SPECT for the investigation of pancreatic cancers. The subjects included 24 patients with pancreatic cancer, seven with benign disorders and 10 controls. Each patient fasted prior to the examination for more than 12 hr, and 201T1 SPECT was obtained 10 min after the injection of 148-222 MBq of 201T1. When the boundary of tumor uptake of 201T1 was unclear because of the adjacent physiological liver activity, subtracted SPECT using 99mTc-phytate was performed to clarify it. 201T1 did not accumulate in the pancreas of the controls. In contrast, of the 24 pancreatic cancers, 21 demonstrated positive uptake, for a sensitivity rate of 87.5%, and the mean tumor/liver ratio was 0.76 +/- 0.16 (range, 0.58-1.28). Abnormal uptake was also noted in three of the seven benign disorders, but with a comparatively lower lesion/liver ratio (range, 0.35-0.51). 201T1 activity per mg tissue in the resected specimens of two patients with pancreatic cancer revealed higher activity in the tumor than in normal parenchyma. 201T1 uptake in the five conservatively treated pancreatic cancers showed alteration similar to the serum level of tumor markers. These results suggest that 201T1 SPECT may have clinical potential for investigating pancreatic cancers as well as for the monitoring of treatment effect. PMID- 7478954 TI - [Palliative radiotherapy for symptomatic osseous metastases]. AB - Bone metastases are one of the most common and serious conditions requiring radiotherapy, but there is still a considerable lack of agreement on optimal radiation schedule. We analyzed patients with symptomatic osseous metastases from lung (72 patients) and breast (63 patients) carcinoma treated by palliative radiotherapy between 1983 and 1992. In this series, the incidences of symptomatic bone metastases appearing within 2 years after the first diagnosis of the primary lesion were 96% and 36% for lung and breast carcinomas, respectively. Thirty percent of bone metastases from breast carcinoma were diagnosed more than 5 years after the first diagnosis. Thus careful follow-up must be carried out for a prolonged period. Pain relief was achieved at almost the same rate for bone metastases from lung and breast carcinomas (81% and 85%, respectively), an the rapid onset of pain relief (15 Gy or less) was obtained in about half the patients for both diseases. The rapid onset of pain relief and the lack of association between the onset of pain relief and primary tumor argued against the conventional theory that tumor shrinkage is a component of the initial response. In contrast to the fact that almost all lung carcinoma patients had very poor prognoses, one third of the breast carcinoma patients were alive more than 2 years after palliative radiotherapy. Thus, the late effects of radiation, such as radiation myelopathy, must be always considered especially in breast carcinoma patients even when it is 'just' palliative radiotherapy for bone metastases. PMID- 7478956 TI - [Radiology reporting system based on the concept of hierarchical structure]. AB - A new radiology reporting system based on the concept of hierarchical structure is proposed. The system generates an intermediate language file made up of semantically linked key words. The hierarchical structure refers to the model of Open System Interconnection (OSI) proposed by the International Organization for Standardization (ISO). The intermediate language is managed and stored along with free sentences (descriptive language) to form a hierarchical structure. Reports of 57 echographies including 65 liver diseases and 7 normal studies done by two medical doctors and two radiologic technicians were tested to evaluate the feasibility and utility of this reporting system. The model system was developed using a personal computer (EPSON PC-486GF) and Japanese word processing software (Justsystem Ichitaro Ver.5/Windows). The hit ratio of the number of picked up keywords (intermediate language) against the number of key findings contained in the free sentences (descriptive language) was 97.4%, and the ratio of hit cases out of total cases was 88.7%. This reporting system with free-text process and hierarchical structure is feasible and useful for improvement of radiological information passage and formation of a radiology database, whereas the strict standardization of descriptive words and criteria of findings will be required for the construction of a practical system. PMID- 7478957 TI - [The use of high concentration ferric ammonium citrate (FAC) solution as a negative bowel contrast agent: application in MR cholangiography]. AB - Ferric ammonium citrate (FAC) can be used as a negative contrast agent on T2- weighted images, with the use of a high concentration resulting in a significant T2 shortening effect. Ten healthy volunteers underwent MR cholangiography (MRC) using Turbo Spin Echo. High-concentration FAC was administered at a normal dosage, but was diluted in a smaller amount of water than usual. Although all precontrast images showed high intensity intestinal fluid, which aggravated visualization of the bile duct, all postcontrast images obtained 6 to 14 minutes after administration showed blackening out of the fluid, improving the overall image quality of MRC. PMID- 7478959 TI - [A threat against public health in Scandinavia]. PMID- 7478958 TI - [Development of apoptosis induced by whole-body irradiation in murine liver]. AB - Apoptosis is known to be induced by radiation. However, the correlation between radiation-induced apoptosis and radiation injury in tumors in vivo has been unclear. In this paper, we report the study of apoptosis induced by whole-body irradiation using an immunohistochemical technique to detect DNA fragmentation in murine liver. A dose of 7 Gy was employed as LD50/30. DNA fragmentation was observed 30 min after radiation, and it peaked between 1 and 6 hours. DNA fragmentation could be detected 48 hours after radiation in capillary endothelium. This study was able to reveal the development of radiation-induced apoptosis in the liver by detecting DNA fragmentation in situ. PMID- 7478960 TI - [Public health services and research in Scandinavia]. AB - Public health problems in the Nordic countries are essentially the same as those in other West European countries: increased inequality in health status, despite the favourable improvement in public health as a whole; the health hazards that accompany increasing unemployment; and the difficulty of maintaining reasonable quality and access to care, owing to diminishing resources. In the Nordic countries, however, there is a broad and diversified collaboration in public health facilities and research based, for instance, on epidemiological studies and assisted by access to health-related data bases. The Nordic countries are characterised by many noteworthy examples of organisational models of public health services at the regional and municipal level. PMID- 7478961 TI - [Scandinavian Public Health Week. Researchers meet practitioners]. PMID- 7478962 TI - [Evaluation of public health services]. AB - The Nordic Congress on Public Health, at Gothenborg, in June 1995, was focused on questions of evaluation. The article reviews basic evaluation concepts, presents a checklist for the planning of evaluation, and proposes an evaluation strategy, as well as standardised terminology for use in trials related to the spread of innovations. The following terms are proposed: news trials, demonstration trials, acceptance trials, and quality assurance trials, each type of trial being linked with appropriate methods. It is suggested that Nordic consensus on this terminology should be achieved as a basis for common language and understanding. PMID- 7478963 TI - [Quality in preventive and health promotion activities]. AB - Quality assurance means ensuring that the right things are done in the right way from the outset and without fail. It is in professional interests to follow up, evaluate and continually attempt to improve preventive and health promotion efforts. In the article an account is given of some concepts, systems, models, methods and working procedures that are used for follow-up, evaluation and development in different fields. Examples are given of the practical applications, above all within the fields of disease prevention and health promotion, booth at the individual and population levels. The aim is to determine the applicability of such endeavours. PMID- 7478965 TI - [Quality assurance in amputation of the lower extremities due to vascular insufficiency]. AB - In this retrospective study we reviewed the clinical charts for 82 patients who underwent lower limb amputation at our department during 1990 and 1991. Our results are compared with the literature, and different aspects of the treatment are discussed. Subsequently a model for future quality assurance is presented. PMID- 7478964 TI - [Vitamin D and osteoporosis]. AB - Vitamin D constitutes a complex endocrine-regulated system, and is both a prohormone for the endogenous synthesis of the active hormone, calcitriol, and a vitamin which may be administered to supply the organism's requirements. No single test or investigation is available for the demonstration of vitamin D deficiency. Both vitamin D intake and ability to synthesise vitamin D decrease with increasing age, and particularly the elderly in institutionalised care are at risk of developing vitamin D deficiency. Iceland excepted, mean daily vitamin D consumption in the Nordic countries is less then 5 micrograms; and in approximately 10-25 per cent of the population, daily intake is less than 2.5 micrograms which is insufficient to maintain an adequate serum calcidiol concentration in individuals unexposed to sunlight. The recommended daily intake of 5 micrograms, currently adopted in the Nordic countries, may be too low-an intake of 10 micrograms is probably necessary to satisfy requirements in the elderly. PMID- 7478966 TI - [Early diagnosis important in hearing loss in children]. AB - Owing to the identification of pre-, peri- and post-natal risk factors for the development of hearing loss or impairment, diagnosis is now possible at 10-14 months of age and rehabilitative measures can be implemented. It is vital that hearing impairment is discovered at an early stage, as language development occurs predominantly before the age of three. Several methods have been evolved for testing hearing in infants. PMID- 7478967 TI - [Time for integrated emergency medicine air ambulance services in Scandinavia?]. AB - The initiative has been taken to develop an integrated system of aeromedical services in the Nordic countries, it is proposed that the system will form a part of public health services, and thus be based on medical rather than commercial criteria. PMID- 7478968 TI - [Telemedicine--a method with great potential development]. PMID- 7478969 TI - [Hospital without borders--visions of telemedicine]. AB - Transfer the expertise rather than the patient! With the help of a screen and a videocamera the general practitioner requiring expert help in diagnosis is able to consult the specialists directly via the broadband ISDN (Integrated Services Digital Network), and referrals complemented with audio-visual material can also be transmitted via the network. Modern telemedicine technology enables hospitals to be linked up, advanced information to be transmitted, interpreter services to be enlisted, and continuous postgraduate training to be developed. Thus, time and money can be saved, the patients given better and faster service, and the quality of health care enhanced. PMID- 7478970 TI - [Patient consultation via broad-band network--pilot project in hand surgery]. AB - A pilot project launched at Esbjerg was designed to ascertain whether broad-band audiovisual transmission of clinical examination can replace direct contact between patient and physician, and if so to what extent. Provided the technique can be improved and simplified, video consultation is an obvious and economically advantageous alternative to referral to a specialised tertiary clinic. Not only will the technique need to be further tested and improved, however, but doctors will require training in the assessment of moving two-dimensional images and diagnosis via a visual display unit (VDU). PMID- 7478971 TI - [Multipoint video-conference. An option for highly specialized education]. AB - Video-conferencing via the PST (Public Switched Telephone) network is a good and economical alternative for use in education. If the universities cooperate, instruction by the best lecturers can be made available to small groups of students distributed over great distances. Travel costs will be minimised and teachers under pressure of time will be able to use their time rationally. Although the social intercourse of a study group promotes the students' mental energy to the benefit of instruction, such technical advances as more rapid transmission and audiovisual integration will be needed. Last but not least, efficient course secretaries will be required to coordinate the work both of lecturers and studio and telephone personnel, as well as the participation of the various study groups. PMID- 7478972 TI - [Heart-related chest pain without manifest coronary disease]. AB - Chest pain is a common reason for consulting a physician. Although the underlying cause of the chest pain varies, the most frequent cause is coronary disease. In many cases the diagnosis emerges from the case history, the clinical picture or non-invasive examination procedures. In cases of severe symptoms or unclear diagnosis, coronary angiography is performed which often shows the coronary arteries to be patent. In a large proportion of these patients the chest pain is of non-cardiac origin. In addition to coronary disease, other heart-related conditions may give rise to chest pain. PMID- 7478974 TI - [Mammography in Norway]. PMID- 7478973 TI - [Need for a long-term program in the follow-up of anti-lipid treatment following coronary artery surgery?]. AB - The positive effects of cholesterol-lowering therapy in coronary artery disease are well recognised. This study, on 99 consecutive coronary artery bypass grafted patients, shows that an intensive one-year follow-up at a secondary prevention specialist clinic significantly improves cholesterol levels compared to conventional follow-up in the primary health care system. However, these positive results are not consistent after a 2.6 year return to the primary health care. Consequently, improved collaboration between hospital and primary health care physicians in the development of structured secondary prevention programmes is essential. PMID- 7478975 TI - [9th Scandinavian Congress in General Medicine. Holistic health--the strong point in primary health care]. PMID- 7478976 TI - [The female in medicine]. PMID- 7478978 TI - [Abolition 2000. A physicians' action against nuclear weapons]. PMID- 7478977 TI - [The male patient in general medicine]. PMID- 7478979 TI - [Eprex: historical record and development]. PMID- 7478980 TI - [Erythropoietin and delayed taking of autologous blood: from physiologic secretion to the rationale for exogenous supplementation]. PMID- 7478981 TI - [Delayed autologous transfusion and erythropoietin]. PMID- 7478982 TI - [Definition of a target population for utilization of recombinant erythropoietin with a view to taking of delayed autologous blood]. PMID- 7478983 TI - [Eprex and daily autologous transfusion]. PMID- 7478984 TI - [Role of autologous transfusion in economy measures for blood in surgery]. PMID- 7478985 TI - [Delayed taking of autologous blood in 1995]. PMID- 7478986 TI - Structural and functional similarities of prokaryotic and eukaryotic DNA polymerase sliding clamps. AB - The remarkable processivity of cellular replicative DNA polymerases derive their tight grip to DNA from a ring-shaped protein that encircles DNA and tethers the polymerase to the chromosome. The crystal structures of prototypical 'sliding clamps' of prokaryotes (beta subunit) and eukaryotes (PCNA) are ring shaped proteins for encircling DNA. Although beta is a dimer and PCNA is a trimer, their structures are nearly superimposable. Even though they are not hexamers, the sliding clamps have a pseudo 6-fold symmetry resulting from three globular domains comprising each beta monomer and two domains comprising each PCNA monomer. These domains have the same chain fold and are nearly identical in three dimensions. The amino acid sequences of 11 beta and 13 PCNA proteins from different organisms have been aligned and studied to gain further insight into the relation between the structure and function of these sliding clamps. Furthermore, a putative embryonic form of PCNA is the size of beta and thus may encircle DNA as a dimer like the prokaryotic clamps. PMID- 7478987 TI - Characterisation of RuvAB-Holliday junction complexes by glycerol gradient sedimentation. AB - The Escherichia coli RuvA and RuvB proteins interact specifically with Holliday junctions to promote ATP-dependent branch migration during genetic recombination and DNA repair. In the work described here, glycerol gradient centrifugation was used to investigate the requirements for the formation of pre-branch migration complexes. Since gradient centrifugation provides a simple and gentle method to analyse relatively unstable protein-DNA complexes, we were able to detect RuvA- and RuvAB-Holliday junction complexes without the need for chemical fixation. Using 35S-labelled RuvA protein and 3H-labelled Holliday junctions, we show that RuvA acts as a helicase accessory factor that loads the RuvB helicase onto the Holliday junction by structure-specific interactions. The resulting complex contained both RuvA and RuvB, as detected by Western blotting using serum raised against RuvA and RuvB. The stoichiometry of binding was estimated to be approximately four RuvA tetramers per junction. Formation of the RuvAB-Holliday junction complex required the presence of divalent metal ions and occurred without the need for ATP. However, the stability of the complex was enhanced by the presence of ATP gamma S, a non-hydrolysable ATP analogue. The data support a model for branch migration in which structure-specific binding of Holliday junctions by RuvA targets the assembly of hexameric RuvB rings on DNA. Specific loading of the RuvB ring helicase by RuvA is likely to be the initial step towards ATP-dependent branch migration. PMID- 7478988 TI - Detection and kinetic studies of triplex formation by oligodeoxynucleotides using real-time biomolecular interaction analysis (BIA). AB - Real-time biomolecular interaction analysis (BIA) has been applied to triplex formation between oligodeoxynucleotides. 5'-Biotinylated oligonucleotides were immobilised on the streptavidin-coated surface of a biosensor chip and subsequently hybridised to their complementary strand. Sequence-specific triplex formation was observed when a suitable third-strand oligopyrimidine was injected over the surface-bound duplex. In addition, a single-stranded oligonucleotide immobilised on the chip surface was able to capture a DNA duplex by triplex recognition. The presence of spermine increases the rate of association between the third strand and immobilised duplex, but at elevated spermine concentrations non-specific association is observed. A preliminary kinetic analysis of triplex formation at pH 5.2 by an 11mer third strand containing thymine, cytosine and uracil is reported. Values for the association and dissociation rate constants were determined to be (1.9 +/- 0.2) x 10(3) M-1 s-1 and (8.1 +/- 1.9) x 10(-5) s 1, respectively. PMID- 7478989 TI - Identity elements of human tRNA(Leu): structural requirements for converting human tRNA(Ser) into a leucine acceptor in vitro. AB - We have previously shown that the exchange of the discriminator base A73 of human tRNA(Leu) for G is alone sufficient to achieve complete loss of leucine acceptance and to create an efficient serine acceptor. The reverse identity switch, however, which was studied using T7 RNA polymerase transcripts of in vitro mutagenized tRNA genes, reveals a far more complex pattern of identity elements for tRNA(Leu). Introduction of the following tRNA(Leu)-specific structures is necessary to transform human tRNA(Ser) into an efficient leucine acceptor: the discriminator base A73, the base pairs C3:G70, A4:U69 and G5:C68 of the acceptor stem, C20a of the DHU loop and the long extra arm. In contrast to tRNA(Ser), human tRNA(Leu) identity requires both the sequence and the correct orientation of the long extra arm, whereas only its orientation is essential for serine identity. PMID- 7478990 TI - Identification of the factors that interact with NCBP, an 80 kDa nuclear cap binding protein. AB - It has been shown that the monomethylated cap structure plays important roles in pre-mRNA splicing and nuclear export of RNA. As a candidate for the factor involved in these nuclear events we have previously purified an 80 kDa nuclear cap binding protein (NCBP) from a HeLa cell nuclear extract and isolated its full length cDNA. In this report, in order to obtain a clue to the cellular functions of NCBP, we attempted to identify a factor(s) that interacts with NCBP. Using the yeast two-hybrid system we isolated three clones from a HeLa cell cDNA library. We designated the proteins encoded by these clones NIPs (NCBP interacting proteins). NIP1 and NIP2 have an RNP consensus-type RNA binding domain, whereas NIP3 contains a unique domain of Arg-Glu or Lys-Glu dipeptide repeats. We also show that NCBP requires NIP1 for binding to the cap structure. Possible roles of NIPs in cap-dependent nuclear processes are discussed. PMID- 7478991 TI - Conversion of mammalian tRNA 3' processing endoribonuclease to four-base recognizing RNA cutters. AB - The spermidine-dependent, sequence-specific endoribonuclease (RNase 65) activities in mammalian cell extracts require both protein and 3' truncated tRNA, species of which direct their substrate sequence specificity. Computer analysis for searching possible base pairing between substrate RNAs and their corresponding 3' truncated tRNA, suggested a unified model for substrate recognition mechanism, in which a four-nucleotide (nt) sequence in the target tRNAs 1 nt upstream of their cleavage site, base pairs with the 5' terminal 4 nt sequence of their corresponding 3' truncated tRNA. This model was supported by experiments with several RNA substrates containing a substituted nucleotide in the target 4 nt sequence. In this model, the tRNA substrates and their corresponding 3' truncated tRNA form a complex resembling a 5' processed tRNA precursor containing a 3' trailer, suggesting that the protein component of RNase 65 is identical to tRNA 3' processing endoribonuclease (3' tRNase). Actually, 3' tRNase purified from pig liver cleaved the target RNAs at the expected sites only in the presence of their corresponding 3' truncated tRNA. These results show that the 3' tRNase can be converted to 4 nt specific RNA cutters using the 3' truncated tRNAs. PMID- 7478992 TI - Dam methylase from Escherichia coli: kinetic studies using modified DNA oligomers: hemimethylated substrates. AB - We have measured steady-state kinetics of the N6-adenine methyltransferase Dam Mtase using as substrates non-selfcomplementary tetradecamer duplexs (d[GCCGGATCTAGACG]-d[CGTCTAGATCC-GGC]) containing the hemimethylated GATC target sequence in one or the other strand and modifications in the GATC target sequence of the complementary strands. Modifications included substitution of guanine by hypoxanthine (I), thymine by uracil (U) or 5-ethyl-uracil (E) and adenine by 2,6 diamino-purine (D). Thermodynamic parameters were obtained from the concentration dependence of the melting temperature (Tm) of the duplexes. Large differences in DNA methylation of duplexes containing single dI for dG substitution of the Dam recognition site were observed compared with the canonical substrate, if the substitution involved the top strand (on the G.C rich side). Substitution in either strand by uracil (dU) or 5-ethyluracil (dE) resulted in small perturbation of the methylation patterns. When 2,6-diamino-purine (dD) replaced the adenine to be methylated, small, but significant methylation was observed. The kinetic parameters of the methylation reaction were compared with the thermodynamic free energies and significant correlation was observed. PMID- 7478993 TI - Picornavirus internal ribosome entry segments: comparison of translation efficiency and the requirements for optimal internal initiation of translation in vitro. AB - On the basis of primary sequence comparisons and secondary structure predictions, picornavirus internal ribosome entry segments (IRESes) have been divided into three groups (entero- and rhinoviruses; cardio- and and aphthoviruses; and hepatitis A virus). Here, we describe a detailed comparison of the ability of IRESes from each group to direct internal initiation of translation in vitro using a single dicistronic mRNA (the only variable being the IRES inserted into the dicistronic region). We studied the influence of various parameters on the capacity of six different picornaviral IRESes, and the non-picornaviral hepatitis C virus IRES, to direct internal initiation of translation: salt concentration, the addition of HeLa cell proteins to rabbit reticulocyte lysate translation reactions, the presence of foot-and-mouth disease virus Lb or human rhinovirus 2A proteinase. On the basis of the characteristics of IRES-driven translation in vitro, the picornaviral IRESes can be classified in a similar manner to when sequence homologies are considered. IRESes from each of the three groups responded differently to all of the parameters tested, indicating that while all of these elements can direct internal ribosome entry, the functional requirements for efficient IRES activity vary dramatically. In the individual optimal conditions for translation initiation, the best IRESes were those from the cardio and aphthoviruses, followed by those from the enteroviruses, which exhibited up to 70% of the efficiency of the EMCV element in directing internal initiation of translation. PMID- 7478994 TI - Tissue-specific regulation of the rabbit 15-lipoxygenase gene in erythroid cells by a transcriptional silencer. AB - The 15-lipoxygenase (lox) gene is expressed in a tissue-specific manner, predominantly in erythroid cells but also in airway epithelial cells and eosinophils. We demonstrate in this report that the 5' flanking DNA of the 15-lox gene contains sequences which down-regulate its activity in a variety of non erythroid cell lines but not in two erythroid cell lines. The element has characteristics of a transcriptional 'silencer' since it functions in both orientations. The main activity of the silencer has been mapped to the first 900 bp of 5' flanking DNA, which contains nine binding sites for a nuclear factor present in non-erythroid cells but not in erythroid cells. These binding sites have similar sequences and multiple copies of the binding sites confer tissue specific down-regulation when attached to a minimal lox promoter fragment. The 5' flanking DNA also contains a cluster of three binding sites for the GATA family of transcription factors. PMID- 7478995 TI - Identification of the yeast nuclear gene for the mitochondrial homologue of bacterial ribosomal protein L16. AB - An open reading frame encoding a member of the L16 family of ribosomal proteins is adjacent to the URA7 gene on the left arm of chromosome II in Saccharomyces cerevisiae. The predicted L16-like polypeptide is basic (pl 11.12), contains 232 amino acids (26.52 kDa) and has 36% amino acid sequence identity to E. coli L16. Immunoblot analysis with polyclonal antibodies to the L16-like polypeptide showed specific cross-reaction with a 22,000 Mr mitochondrial polypeptide that co sediments with the large subunit of the mitochondrial ribosome in sucrose density gradients. The levels of the L16 mRNA and protein varied in response to carbon source. In [rho degree] cells lacking mitochondrial rRNA, the L16 mRNA accumulated at normal levels, but the protein was barely detectable, indicating RNA-dependent accumulation of the L16 protein. Gene disruption experiments demonstrated that the yeast mitochondrial L16 is an essential ribosomal protein in vivo. PMID- 7478996 TI - Variability in DNA minor groove width recognised by ligand binding: the crystal structure of a bis-benzimidazole compound bound to the DNA duplex d(CGCGAATTCGCG)2. AB - An analogue of the DNA-binding compound Hoechst 33258, in which the piperazine ring has been replaced by an imidazoline group, has been cocrystallized with the dodecanucleotide sequence d(CGCGAATTCGCG)2. The structure has been solved by X ray diffraction analysis and has been refined to an R-factor of 19.7% at a resolution of 2.0 A. The ligand is found to bind in the minor groove, at the central four AATT base pairs of the B-DNA double helix, with the involvement of a number of van der Waals contacts and hydrogen bonds. There are significant differences in minor groove width for the two compounds, along much of the AATT region. In particular this structure shows a narrower groove at the 3' end of the binding site consistent with the narrower cross-section of the imidazole group compared with the piperazine ring of Hoechst 33258 and therefore a smaller perturbation in groove width. The higher binding affinity to DNA shown by this analogue compared with Hoechst 33258 itself, has been rationalised in terms of these differences. PMID- 7478997 TI - Description of the entire mRNA population by a 3' end cDNA fragment generated by class IIS restriction enzymes. AB - A novel means of recording the expression status of the total gene population is described. Digestion of cDNA by class IIS restriction enzymes produces a fragment with a poly (A) stretch and a 5' overhang with an unknown sequence. This fragment contains information such as the class IIS enzyme that cuts cDNA nearest to the poly (A) stretch, the sequence of the 5' overhang, and the size of the fragment. Expressed genes can be discriminated and displayed by the fragment as follows: (i) cut the cDNA with one class IIS restriction enzyme; (ii) ligate the digested cDNA to that from a pool of 64 biotinylated adaptors cohesive to all possible overhangs; (iii) digest by other two class IIS enzymes; (iv) recover the ligated molecules with streptavidin-coated paramagnetic beads; (v) perform PCR with the adaptor-primer and an anchored oligo-dT primer; (vi) separate the amplified fragments by denaturing polyacrylamide gel electrophoresis. Repeat the experiment with 64 adaptors, three enzymes and three anchored oligo-dT primers displays most of the expressed genes. Because redundancy is minimized, this technique is also ideal for generating tags for expressed genes, with which to construct a transcript map of the genome. PMID- 7478998 TI - Unequal homologous recombination of human DNA on a yeast artificial chromosome. AB - We examined unequal homologous DNA recombination between human repetitive DNA elements located on a yeast artificial chromosome (YAC) and transforming plasmid molecules. A plasmid vector containing an Alu element, as well as a sequence identical to a unique site on a YAC, was introduced into yeast and double recombinant clones analyzed. Recombination occurs between vector and YAC Alu elements sharing as little as 74% identity. The physical proximity of an Alu element to the unique DNA segment appears to play a significant role in determining the frequency with which that element serves as a recombination substrate. In addition, cross-over points of the recombination reaction are largely confined to the ends of the repetitive element. Since a similar distribution of crossover sites occurs during unequal homologous recombination in human germ and somatic tissue, we propose that similar enzymatic processes may be responsible for the events observed in our system and in human cells. This suggests that further examination of the enzymology of unequal homologous recombination of human DNA within yeast may yield a greater understanding of the molecular events which control this process in higher eukaryotes. PMID- 7478999 TI - Computer assisted identification and classification of streptomycete promoters. AB - Short sequences that were over represented in a database of Streptomyces promoter region sequences were identified. These sequences and others that were selected on the basis of the characteristics of known promoters, were tested to determine if they were found predominantly at particular distances from the transcription start site. In several cases obvious clusters were recorded. This has allowed the objective identification of potential promoter core sequences. In some cases these may define novel promoter classes. 150 Streptomyces promoters have been listed and grouped on this basis. A new and extended consensus sequence for the Streptomyces E.coli sigma 70-like promoters was determined. It showed differences from that of E.coli, both in sequence and in the spacing between the -35 and -10 regions. PMID- 7479000 TI - RNA polymerase III dependence of the human L1 promoter and possible participation of the RNA polymerase II factor YY1 in the RNA polymerase III transcription system. AB - From the general views of the eukaryotic transcription systems, L1 (or L1-like) retrotransposons that encode some proteins are unusual. L1, unlike other protein coding elements, is transcribed through an internal promoter. And the L1 internal promoter, unlike other internal promoters, is thought to be RNA polymerase II (pol II) dependent, because the L1 transcript has a large size (approximately 6 kb), protein coding capacity and a 3' terminal polyadenylation signal followed by a poly(A) tail, and also because transcription from the promoter of Drosophila L1 like element jockey was highly sensitive to alpha-amanitin. However, our in vitro transcription study reveals that transcription from the human L1 promoter is highly sensitive to tagetitoxin, a selective inhibitor of RNA polymerase III (pol III), but insensitive to 1 micrograms/ml of alpha-amanitin, indicating that the human L1 promoter is pol III-dependent. The pol III dependence is further supported by our observation that L1 and pol III-dependent tRNA gene promoters share a common nuclear factor YY1. There is evidence that YY1 is also a pol II transcription factor. We thus propose that YY1 is a possible member of the pol III transcription system. PMID- 7479002 TI - The influence of a metastable structure in plasmid primer RNA on antisense RNA binding kinetics. AB - Replication of the ColE1 group plasmids is kinetically regulated by the interaction between plasmid-encoded primer RNA II and antisense RNA I. The binding is dependent on alternative RNA II conformations, formed during the transcription, and effectively inhibits the primer function within some time interval. In this paper, the folding pathways for the wild type and copy number mutants of ColE1 RNA II are studied using simulations by a genetic algorithm. The simulated pathways reveal a transient formation of a metastable structure, which is stabilized by copy number mutations. The folding kinetics of the proposed conformational transitions is calculated using a model of a multistep refolding process with elementary steps of double-helical stem formation or disruption. The approximation shows that the lifetime of the metastable structure is relatively long and is considerably increased in the mutants, resulting in a delay of the formation of the stable RNA II structure, which is the most sensitive to the inhibition by the antisense RNA I. Thus the effect of copy number mutations can be interpreted as a compression of the time window of effective inhibition due to an increased time spent by the RNA II in the metastable state. The implications of metastable foldings in RNA functioning are discussed. PMID- 7479001 TI - Differential regulation of plasminogen activator and inhibitor gene transcription by the tumor suppressor p53. AB - The ability of p53 to activate or repress transcription suggests that its biological function as tumor suppressor is in part accomplished by regulating a number of genes including such required for inhibition of cell growth. We here give evidence that p53 also may regulate genes responsible for the proteolytic degradation of the extracellular matrix, which is considered a crucial feature for local invasion and metastasis of neoplastic cells. An important and highly regulated cascade of such proteolytic events involves the plasminogen activator system. We show that wild-type p53 represses transcription from the enhancer and promoter of the human urokinase-type (u-PA) and the tissue-type plasminogen activator (t-PA) gene through a non-DNA binding mechanism. Oncogenic mutants lost the repressing activity. In contrast, wild-type but not mutant p53 specifically binds to and activates the promoter of the plasminogen activator inhibitor type-1 (PAI-1) gene. Interestingly, one of the p53 mutants (273his) inhibited PAI-1 promoter activity. Our results suggest that altered function of oncogenic forms of p53 may lead to altered expression of the plasminogen activators and their inhibitor(s) and thus to altered activation of the plasminogen/plasmin system during tumor progression. PMID- 7479003 TI - 5'-Cholesteryl-phosphorothioate oligodeoxynucleotides: potent inhibition of methotrexate transport and antagonism of methotrexate toxicity in cells containing the reduced-folate carrier. AB - Polyanionic 5'-cholesteryl-phosphorothioate oligodeoxynucleotides of varying polymer length and nucleobase composition were examined for an effect on methotrexate transport via the reduced-folate carrier of L1210 mouse cells. Methotrexate transport was inhibited by each of the oligodeoxynucleotide analogs tested. Inhibition was most pronounced (IC50 = 0.21 microM, standard assay) for a 5'-cholesteryl heteropolymer consisting of 15 phosphorothioate deoxynucleotides with alternating deoxycytosine and deoxyadenosine (Chol-PS-d(CA)7C). Homopolymers with 15 deoxycytosine (Chol-PS-dC15) or deoxythymidine (Chol-PS-dT15) residues were approximately 2-fold less inhibitory than Chol-PS-d(CA)7C. The relative potency of transport inhibition by deoxycytosine oligomers of varying length was: Chol-PS-dC5 > Chol-PS-dC15 > Chol-PS-dC28 > Chol-PS-dC3. Substantial inhibition was retained in cells preincubated with inhibitors and washed prior to transport determinations and the inhibitor sensitivity could be increased substantially by reducing the concentration of cells. Mixed competitive and non-competitive inhibition was observed for each analog. In standard high-folate medium, Chol-PS oligodeoxynucleotides (5.0 microM) had minimal effects on the growth of L1210 cells, but antagonized the cytotoxicity of methotrexate. The response to methotrexate (IC50 = 12 nM) decreased to the greatest extent (20.8-fold) in the presence of Chol-PS-d(CA)7C (IC50 = 250 nM). Under limiting folate conditions, Chol-PS-d(CA)7C alone inhibited cells growth by a process which could be reversed by folic acid. The results show that Chol-PS-oligodeoxynucleotides are among the most potent known inhibitors of the reduced-folate carrier. Direct growth inhibition of folate-deficient cells and antagonism of methotrexate cytotoxicity indicate that Chol-PS-oligodeoxynucleotides retain the ability to inhibit the reduced-folate carrier for several days in cultured cells. PMID- 7479004 TI - Transcription factors RFX1/EF-C and ATF-1 associate with the adenovirus E1A responsive element of the human proliferating cell nuclear antigen promoter. AB - The proliferating cell nuclear antigen (PCNA) is an adenovirus E1A-inducible factor that is intimately linked to the processes of DNA replication and cell cycle regulation. Previously, we defined a novel cis-acting element, the PCNA E1A responsive element (PERE), that confers induction by the E1A 243R oncoprotein upon the human PCNA promoter. To better understand the regulation of PCNA expression by E1A 243R, we have identified cellular transcription factors that associate with the PERE. In electrophoretic mobility shift assays, the PERE formed three major complexes (P1, P2 and P3) with proteins in nuclear extracts from HeLa or 293 cells. Formation of complexes P2 and P3, which correlates with PCNA promoter activity in vivo, requires the activating transcription factor (ATF) binding site found within the PERE [Labrie et al. (1993) Mol. Cell. Biol., 13, 1697-1707]. Antibody interference experiments and mobility shift assays performed with in vitro-synthesized protein indicated that the transcription factor ATF-1 is a major component of these complexes. Similar assays demonstrated that the hepatitis B virus enhancer-associated protein RFX1 constitutes a major component of the P1 complex. In addition, we examined the binding of proteins to the minimal E1A-responsive promoter to identify other factors important for transcription from the PCNA promoter. Mobility shift assays revealed that a fragment encompassing the region from -87 to +62 relative to the transcription initiation site forms at least five complexes, EH1-EH5, with HeLa cell nuclear extracts. The transcription factor YY1 associates with the initiator element of the PCNA promoter. The identification of these transcription factors will allow their roles in the activation of PCNA by E1A to be evaluated. PMID- 7479006 TI - Escherichia coli, Saccharomyces cerevisiae, rat and human 3-methyladenine DNA glycosylases repair 1,N6-ethenoadenine when present in DNA. AB - The human carcinogen vinyl chloride is metabolized in the liver to reactive intermediates which generate various ethenobases in DNA. It has been reported that 1,N6-ethenoadenine (epsilon A) is excised by a DNA glycosylase present in human cell extracts, whereas protein extracts from Escherichia coli and yeast were devoid of such an activity. We confirm that the human 3-methyladenine-DNA glycosylase (ANPG protein) excises epsilon A residues. This finding was extended to the rat (ADPG protein). We show, at variance with the previous report, that pure E.coli 3-methyladenine-DNA glycosylase II (AlkA protein) as well as its yeast counterpart, the MAG protein, excise epsilon A from double stranded oligodeoxynucleotides that contain a single epsilon A. Both enzymes act as DNA glycosylases. The full length and the truncated human (ANPG 70 and 40 proteins, respectively) and the rat (ADPG protein) 3-methyladenine-DNA glycosylases activities towards epsilon A are 2-3 orders of magnitude more efficient than the E.coli or yeast enzyme for the removal of epsilon A. The Km of the various proteins were measured. They are 24, 200 and 800 nM for the ANPG, MAG and AlkA proteins respectively. These three proteins efficiently cleave duplex oligonucleotides containing epsilon A positioned opposite T, G, C or epsilon A. However the MAG protein excises A opposite cytosine much faster than opposite thymine, guanine or adenine. PMID- 7479005 TI - Enzymatic synthesis of polymers containing nicotinamide mononucleotide. AB - Nicotinamide mononucleoside 5'-diphosphate in its reduced form is an excellent substrate for polynucleotide phosphorylase from Micrococcus luteus both in de novo polymerization reactions and in primer extension reactions. The oxidized form of the diphosphate is a much less efficient substrate; it can be used to extend primers but does not oligomerize in the absence of a primer. The cyanide adduct of the oxidized substrate, like the reduced substrate, polymerizes efficiently. Loss of cyanide yields high molecular weight polymers of the oxidized form. Terminal transferase from calf thymus accepts nicotinamide mononucleoside 5'-triphosphate as a substrate and efficiently adds one residue to the 3'-end of an oligodeoxynucleotide. T4 polynucleotide kinase accepts oligomers of nicotinamide mononucleotide as substrates. However, RNA polymerases do not incorporate nicotinamide mononucleoside 5'-triphosphate into products on any of the templates that we used. PMID- 7479007 TI - The uni chromosome of Chlamydomonas: histone genes and nucleosome structure. AB - The uni linkage group (ULG) of Chlamydomonas reinhardtii contains many genes involved in the basal body-flagellar system. Recent evidence suggests that the corresponding uni chromosome is located in close proximity to the basal body complex. In the course of studies into its molecular organization, we have found a cluster of four histone genes on the ULG. The genes are arranged as divergently transcribed pairs: H3-H4 and H2B-H2A. Genomic sequencing reveals that these genes lack introns and contain characteristic 3' palindromes similar to those of animals. The predicted amino acid sequences are highly conserved across species, with greatest similarities to the histone genes of Volvox. Southern analysis shows that each histone gene is present in 15-20 copies in Chlamydomonas and suggests a dispersed genomic organization. Northern analysis of mitotically synchronized cells shows that, like the replication-dependent histones of higher eukaryotes, Chlamydomonas histone genes are expressed during S-phase. Using a gene-specific probe on Northern blots, we provide evidence that the ULG H4 gene is regulated in the same manner as other Chlamydomonas histone genes. Finally, micrococcal nuclease protection experiments show that the uni chromosome itself associates with histone proteins and displays a conventional nucleosomal banding pattern. PMID- 7479008 TI - Replacing tryptophan-128 of T4 endonuclease V with a serine residue results in decreased enzymatic activity in vitro and in vivo. AB - Endonuclease V of bacteriophage T4 possesses two enzymatic activities, a DNA N glycosylase specific for cyclobutane pyrimidine dimers (CBPD) and an associated apurinic/apyrimidinic (AP) lyase. Extensive structural and functional studies of endonuclease V have revealed that specific amino acids are associated with these two activities. Controversy still exists regarding the role of the aromatic amino acid stretch close to the carboxyl terminus, in particular the tryptophan at position 128. We have expressed wild-type and mutant W128S endonuclease V in Escherichia coli from an inducible tac promoter. Purified W128S endonuclease V demonstrated substantially decreased N-glycosylase (approximately 5-fold) and AP lyase (10- to 20-fold) activities in vitro compared to the wild-type enzyme when a UV-irradiated poly(dA)-poly(dT) substrate was used. However, a much smaller difference in AP lyase activity between the two forms was observed with a site specific abasic oligonucleotide. The difference in enzymatic activity was qualitatively, but not quantitatively, reflected in the survival of UV-irradiated bacteria, that is the W128S cells were slightly less UV resistant than wild-type cells. No difference was observed in the complementation of UV repair using UV damaged denV- T4 phage. A more pronounced difference between the wild-type and W128S proteins was observed in human xeroderma pigmentosum group A cells by host cell reactivation of a UV-irradiated reporter gene. The relatively large discrepancy between the in vitro and in vivo results observed with bacteria may be because saturated levels of DNA repair are obtained in vivo with relatively low levels of endonuclease V. However, under limiting in vitro conditions and in human cells in vivo a considerable difference between the W128S mutant and wild type endonuclease V activities can be detected. Our results demonstrate that tryptophan-128 is important for endonuclease V activity. PMID- 7479009 TI - Formation of a parallel-stranded DNA homoduplex by d(GGA) repeat oligonucleotides. AB - The GGA9-H molecules consisting of a double helical stretch followed by a single stranded 3'-terminal overhang of nine GGA sequence repeats exhibited a gel mobility-shifted band in a concentration-dependent manner, suggestive of the intermolecular complex formation. The position of the shifted band in a gel was almost identical to that of the Y-shaped dimer marker of the same molecular weight that had the two double-helices at one side. This suggests that GGA9-H dimerizes in a parallel orientation without the formation of four-stranded hairpin structure. Since the GGA9-H homoduplex was stably formed at pH 4, 7 and 9, the formation does not require protonation or deprotonation of the N1 position of adenines. Neither does it require the N7 group of guanines responsible for Hoogsteen base pairing from the methylation interference and modification studies. Modification of the N7 group of guanines with dimethyl sulfate (DMS) did not inhibit the association and also the N7 group in the homoduplex was not protected from DMS. On the other hand, the GAA9-H having the G to A base substitution did not show such an association with either GGA9-H or GAA9-H. These results suggest that the homoduplex formation may be due to G.G base pairing through non-Hoogsteen hydrogen bonds. PMID- 7479011 TI - Chromatin structure snap-shots: rapid nuclease digestion of chromatin in yeast. PMID- 7479010 TI - Binding site requirements and differential representation of TGF factors in nuclear ASF-1 activity. AB - Activating sequence factor 1 (ASF-1) is a nuclear DNA-binding activity that is found in monocots and dicots. It interacts with several TGACG-containing elements that have been characterized from viral and T-DNA genes, the prototypes of which are the as-1 element of the CaMV 35S promoter and the ocs element from the octopine synthase promoter. This class of cis-acting elements can respond to auxin and salicylic acid treatments. Consistent with these observations, we have shown that ASF-1 can interact with promoter elements of an auxin-inducible tobacco gene GNT35, encoding a glutathione S-transferase. Characterization of the nuclear factors that make up ASF-1 activity in vivo will be an important step toward understanding this induction phenomenon. The TGA family of basic-leucine zipper (bZIP) proteins are good candidates for the ASF-1 nuclear factor. However, there may be as many as seven distinct TGA genes in Arabidopsis, five of which have now been reported. In this study, we expressed the cDNAs that encode four of these five Arabidopsis TGA factors in vitro and compared their DNA-binding behavior using two types of TGACG-containing elements. With specific antisera prepared against three of the five known Arabidopsis TGA factors, we also investigated the relative abundance of these three proteins within the ASF-1 activities of root and leaf nuclear extracts. Our results indicate that these TGA factors bind to DNA with different degrees of cooperativity and their relative affinity toward as-1 also can differ significantly. The results of a supershift assay suggested that only one of the three TGA factors represented a significant component of nuclear ASF-1 activity. Arabidopsis TGA2 comprises approximately 33 and 50% of the ASF-1 activity detected in root and leaf nuclear extracts respectively. These results suggest that each member of the TGA factor family may be differentially regulated and that they may play different roles by virtue of their distinct DNA-binding characteristics. Furthermore, since transcripts for each of these factors can be detected in various plant tissues, post transcriptional regulation may play an important part in determining their contribution to nuclear ASF-1 in a given cell type. PMID- 7479012 TI - Enhanced direct amplification of Guthrie card DNA following selective elution of PCR inhibitors. PMID- 7479013 TI - Differentiation of the normal and mutant rat albumin genes on hepatic tissue sections by in situ PCR. PMID- 7479015 TI - Modifications of a telomeric repeat amplification protocol (TRAP) result in increased reliability, linearity and sensitivity. PMID- 7479014 TI - Biotinylated probes in the electrophoretic mobility shift assay to examine specific dsDNA, ssDNA or RNA-protein interactions. PMID- 7479016 TI - cRACE: a simple method for identification of the 5' end of mRNAs. PMID- 7479017 TI - Hybridization of microsatellites to RAPD: a new source of polymorphic markers. PMID- 7479019 TI - Random amplified hybridization microsatellites (RAHM): isolation of a new class of microsatellite-containing DNA clones. PMID- 7479018 TI - A method for preparation of fecal DNA suitable for PCR. PMID- 7479020 TI - Induction of double-strand breaks by S1 nuclease, mung bean nuclease and nuclease P1 in DNA containing abasic sites and nicks. AB - Defined DNA substrates containing discrete abasic sites or paired abasic sites set 1, 3, 5 and 7 bases apart on opposite strands were constructed to examine the reactivity of S1, mung bean and P1 nucleases towards abasic sites. None of the enzymes acted on the substrate containing discrete abasic sites. Under conditions where little or no non-specific DNA degradation was observed, all three nucleases were able to generate double-strand breaks when the bistranded abasic sites were 1 and 3 base pairs apart. However, when the abasic sites were further apart, the enzymes again failed to cleave the DNA. These results indicate that single abasic sites do not cause sufficient denaturation of the DNA to allow incision by these single-strand specific endonucleases. The reactivity of these enzymes was also investigated on DNA substrates that were nicked by DNasel or more site specifically by endonuclease III incision at the discrete abasic sites. The three nucleases readily induced a strand break opposite such nicks. PMID- 7479021 TI - Strategic down-regulation of DNA polymerase beta by antisense RNA sensitizes mammalian cells to specific DNA damaging agents. AB - Previously, mouse NIH 3T3 cells were stably transfected with human DNA polymerase beta (beta-pol) cDNA in the antisense orientation and under the control of a metallothionein promoter [Zmudzka, B.Z. and Wilson, S.H. (1990) Som. Cell Mol. Gen., 16, 311-320]. To assess the feasibility of enhancing the efficacy of chemotherapy by an antisense approach and to confirm a role for beta-pol in cellular DNA repair, we looked for increased sensitivity to DNA damaging agents under conditions where beta-pol is down-regulated in the antisense cell line. Such a sensitization is anticipated only where beta-pol is rate-limiting in a DNA repair pathway. A number of agents were tested: cis-diamminedichloroplatinum II (cisplatin); 1,3-bis(2-chloroethyl)-1- nitrosourea (BCNU); ionizing radiation and the radio-mimetic drug bleomycin; the bifunctional alkylating agents nitrogen mustard and L-phenylalanine mustard (melphalan); the monofunctional alkylating agent methyl methane sulfonate (MMS) and ultraviolet (UV) radiation. In the cases of cisplatin and UV radiation, a significant enhancement of cytotoxicity was observed. Damage as a result of both of these agents is thought to be repaired by the nucleotide excision repair (NER) pathway. The results suggest that, in this cell line, beta-pol is involved in and is rate-limiting in NER. We propose that down-regulation of beta-pol by antisense approaches might be used to enhance the cytotoxic effects of cisplatin and other DNA damaging chemotherapeutic agents. PMID- 7479022 TI - Efficient gene activation in mammalian cells by using recombinant adenovirus expressing site-specific Cre recombinase. AB - A recombinant adenovirus (Ad) expressing Cre recombinase derived from bacteriophage P1 was constructed. To assay the Cre activity in mammalian cells, another recombinant Ad bearing an on/off-switching reporter unit, where a LacZ expression unit can be activated by the Cre-mediated excisional deletion of an interposed stuffer DNA, was also constructed. Co-infection experiments together with the Cre-expressing and the reporter recombinant Ads showed that the Cre mediated switching of gene expression was detected in nearly 100% of cultured CV1, HeLa and Jurkat cells. These results suggest that the recombinant Ad efficiently expressed functional Cre and offers a basis for establishing a powerful on/off switching strategy of gene expression in cultured mammalian cells and presumably in transgenic animals. The method is also applicable to construction of recombinant Ad bearing a gene the expression of which is deleterious to propagation of recombinant Ad. PMID- 7479023 TI - Cell cycle regulation of cdc25C transcription is mediated by the periodic repression of the glutamine-rich activators NF-Y and Sp1. AB - The late S/G2-specific transcription of the human cdc25C gene is dependent on an initiator-proximal repressor element (CDE) and an upstream activating sequence (UAS) of undefined nature. We now show that these upstream sequences harbour multiple in vivo protein binding sites that interact with transcriptional activators and form separable, context-independent functional modules. Major components of the UAS are a bona fide Sp1 site and three direct sequence repeats (Yc-boxes). The Yc-boxes interact with the CCAAT-box binding protein NF-Y and are critically dependent on synergistic interactions for efficient transcription activation. The NF-Y complexes, as well as Sp1, are constitutive activators, whose activation function is periodically repressed through the CDE. These observations indicate that the cell cycle regulation of cdc25C transcription is mainly due to the CDE-mediated repression of glutamine-rich activators. PMID- 7479024 TI - The high stability of the triple helices formed between short purine oligonucleotides and SIV/HIV-2 vpx genes is determined by the targeted DNA structure. AB - In our previous works we have shown that the oligonucleotides 5'-GGGGAGGGGGAGG-3' and 5'-GGAGGGGGAGGGG-3' give very stable and specific triplexes with their target double stranded DNAs [Svinarchuk, F., Bertrand, J.-R. and Malvy, C. (1994) Nucleic Acids Res., 22, 3742-3747; Svinarchuk, F., Paoletti, J. and Malvy, C. (1995) J. Biol. Chem., 270, 14 068-14,071]. The target for the invariable part of these oligonucleotides, 5'-GGAGGGGGAGG-3', is found in a highly conserved 20 bp long purine/pyrimidine tract of the vpx gene of the SIV and HIV-2 viruses and could be a target for oligonucleotide directed antivirus therapy. Here were report on the ability of four purine oligonucleotides with different lengths (11 , 14-, 17- and 20-mer) to form triplexes with the purine/pyrimidine stretch of the vpx gene. Triplex formation was tested by joint dimethyl sulfate (DMS) footprint, gel-retardation assay, circular dichroism (CD) and UV-melting studies. Dimethyl sulfate footprint studies revealed the antiparallel orientation of the third strand to the purine strand of the Watson-Crick duplex. However, the protection of the guanines at the ends of the target sequence decreased as the length of the third strand oligonucleotide increased. Melting temperature studies provided profiles with only one transition for all of the triplexes. The melting temperatures of the triplexes were found to be the same as for the targeted duplex in the case of the 11- and 14-mer third strands while for the 17- and 20 mer third strands the melting temperature of the triplexes were correspondingly 4 and 8 degrees C higher than for the duplex. Heating and cooling melting curves were reversible for all of the tested triplexes except one with the 20-mer third strand oligonucleotide. Circular dichroism spectra showed the ability of the target DNA to adopt an A-like DNA conformation. Upon triplex formation the A-DNA form becomes even more pronounced. This effect depends on the length of the third strand oligonucleotide: the CD spectrum shows a 'classical' A-DNA shape with the 20-mer. This is not observed with the purine/pyrimidine stretch of the HIV-1 DNA which keeps a B-like spectrum even after triplex formation. We suggest, that an A like duplex DNA is required for the formation of a stable DNA purine(purine pyrimidine) triplex. PMID- 7479025 TI - Promoter recognition in archaea is mediated by transcription factors: identification of transcription factor aTFB from Methanococcus thermolithotrophicus as archaeal TATA-binding protein. AB - At least two transcription factors, aTFB and aTFA, are required for accurate and faithful in vitro transcription of homologous templates in cell-free extracts from the methanogenic Archaeon Methanococcus thermolithotrophicus. We have recently shown that the function of aTFB can be replaced by eucaryal TATA-binding proteins. Here we demonstrate using template commitment experiments that promoter recognition in an Archaeon is mediated by transcription factors. The archaeal TATA box was identified as recognition site for binding of aTFB by gel shift analyses. aTFB binds also to the TATA box of adenovirus 2 major late promoter suggesting homology of eucaryal and archaeal TATA boxes. Our analyses provide evidence for a common molecular mechanism of transcription initiation by eucaryal RNA polymerases and archaeal RNA polymerase. They indicate also an evolutionary homology for aTFB and TBP. PMID- 7479027 TI - Hairpins in a DNA site for topoisomerase II studied by 1H- and 31P-NMR. AB - 1H- and 31P-NMR and UV-absorption studies were carried out with the oligonucleotide strands d(AGCT-TATC-ATC-GATAAGCT) (-ATC-) and d(AGCTTATC-GAT GATAAGCT) (-GAT-) contained in the strongest and salt resistant cleavage site for topoisomerase II in pBR322 DNA. We found that the two oligonucleotides were stabilized under a hairpin structure characterized by a eight base pair stem and a three base loop at low DNA and salt concentrations. In such experimental conditions, only the -GAT- oligonucleotide displayed a partial homoduplex structure in slow equilibrium with its folded structure. Temperature dependencies of imino protons showed that the partial homoduplex of -GAT- melted at a lower temperature than the hairpin structure. It was suggested that the appearance of the partial homoduplex in -GAT- is related to the formation of two stabilizing (G.T) mismatched base pairs in the central loop of this structure. Finally, it was inferred from the dispersion of chemical shifts in the 31P-NMR spectra that the distortions affecting the backbone of the hairpin loop are larger in the case of -ATC- compared with -GAT-. At the same time NOEs proved that the base stacking was stronger within the loop of the -ATC- hairpin. PMID- 7479026 TI - Fine-mapping of shotgun template-libraries; an efficient strategy for the systematic sequencing of genomic DNA. AB - To test the effectiveness of ordering shotgun DNA-templates prior to sequence analysis, the 450 kb left arm of yeast chromosome XII was randomly subcloned into a phagemid vector. Clones were ordered by hybridisation to an average map density of one new insert every 125 bp and are currently used for sequencing the chromosomal fragment. An 11.5 kb overlap between the template map and a DNA fragment that had been sequenced earlier allowed an independent evaluation of the strategy's effectiveness. To this end, clones were selected from the map and tag sequenced from either end, thus comparing the map position with the actual location within the 11.5 kb. Of 65 selected clones, taken mostly at random from a total of 423, 58 mapped on average about a quarter of a clone length around their predicted position, with the other seven being between 0.6 and 1.5 clone length off. 75-86 sequencing reactions on clones selected from the map would have been sufficient for completely sequencing both strands of the 11.5 kb fragment. The results demonstrate the efficacy of such template sorting, considerably assisting sequencing at relatively little cost on the mapping level. PMID- 7479029 TI - TFE3 contains two activation domains, one acidic and the other proline-rich, that synergistically activate transcription. AB - TFE3 is a basic-helix-loop-helix-zipper (bHLHZIP) domain-containing protein that binds mu E3 sites in regulatory elements in the immunoglobulin heavy chain gene. The protein is a transcriptional activator that is expressed in vivo as two alternately spliced isoforms with different activating properties: TFE3L contains an N-terminal acidic activation domain; TFE3S lacks this activation domain and is a dominant negative inhibitor of TFE3L. We show that TFE3L and TFE3S contain a second, C-terminal activation domain rich in proline residues. This pro-rich activation domain has activity in a Gal4 fusion assay comparable to the N terminal acidic activation domain present in TFE3L. The TFE3 pro-rich activation domain contains regions of strong homology with the related proteins microphthalmia and TFEB, suggesting that these regions are important for function. Using two different assays, we show that the N- and C-terminal activation domains of TFE3 act synergistically. This synergism explains in part the ability of TFE3S to act as a dominant negative. Our domain analysis of TFE3 is incorporated into a general structural model for the TFE3 protein that predicts that the activation domains of TFE3 will be widely separated in space. PMID- 7479028 TI - A human NDP-kinase B specifically binds single-stranded poly-pyrimidine sequences. AB - Recently, a DNA binding protein 'PUF' was purified that binds to a poly pyrimidine rich element in the human c-myc promoter. Cloning of the corresponding gene surprisingly identified this putative transcription factor as isoform B of the enzyme nucleoside diphosphate kinase (NDPK-B) [Postel et al. (1993) Science, 261, 478-480], the product of the potential metastasis suppressor gene nm23-H2. Using different recombinant NDP kinases, we demonstrate by electrophoretic mobility shift analysis (EMSA) that the NDP kinase DNA binding properties are predominantly observed with human isoform B. Unlike typical DNA binding proteins that are involved in transcriptional regulation, binding occurs to single stranded DNA rather than to a double-stranded oligonucleotide. As a consequence, complexes of single-stranded DNA and NDPK-B are generated from double-stranded oligonucleotide hybrids in an ATP independent manner. In addition to the c-myc element, NDPK-B is binding in vitro to a variety of poly-pyrimidine rich sequences including dC or dT homo-oligomers, (CT)n dinucleotide repeats, the initiator region of the Adenovirus major late promoter and even poly-pyrimidine rich RNAs. The possible consequences of these findings in understanding the multiple roles of NDP kinase are discussed. PMID- 7479030 TI - Hairpin opening by single-strand-specific nucleases. AB - DNA molecules with covalently sealed (hairpin) ends are probable intermediates in V(D)J recombination. According to current models hairpin ends are opened to produce short single-stranded extensions that are thought to be precursors of a particular type of extra nucleotides, termed P nucleotides, which are frequently present at recombination junctions. Nothing is known about the activities responsible for hairpin opening. We have used two single-strand-specific nucleases to explore the effects of loop sequence on the hairpin opening reaction. Here we show that a variety of hairpin ends are opened by P1 nuclease and mung bean nuclease (MBN) to leave short, 1-2 nt single-stranded extensions. Analysis of 22 different hairpin sequences demonstrates that the terminal 4 nt of the hairpin loop strongly influence the sites of cleavage. Correlation of the nuclease digestion patterns with structural (NMR) data for some of the hairpin loops studied here provides new insights into the structural features recognized by these enzymes. PMID- 7479031 TI - Microsatellite variation in North American populations of Drosophila melanogaster. AB - Computer database searching for microsatellites can be particularly effective for organisms like Drosophila melanogaster for which there are extensive sequence data. Here we demonstrate that 17 out of 18 such microsatellites are also highly polymorphic in natural populations of Drosophila, and that this variation is easily scorable with PCR followed by electrophoresis on high-resolution agarose. This form of variation is likely to be of great value in studies of the genomic distribution of polymorphism, population structure, the relation between intraspecific polymorphism and interspecific divergence and the mutation rate and pattern of mutations of microsatellites. In this preliminary survey of 15 lines, we find that the variance in repeat count is most strongly correlated with the maximum count, that perfect repeats are significantly more variable than imperfect repeats and that repeats which are split by an imperfection have unexpectedly low variance given the size of the perfectly repeated portion. PMID- 7479032 TI - Inhibition of NF-kappa B-Rel A expression by antisense oligodeoxynucleotides suppresses synthesis of urokinase-type plasminogen activator (uPA) but not its inhibitor PAI-1. AB - The essential role of urokinase-type plasminogen activator (uPA) in tumor invasion and metastasis stresses the necessity of a fine-tuned cellular control over its expression. It has been shown that changes in uPA directly correlate with changes in cell invasiveness. We examined the role of Rel-related proteins in uPA synthesis by human ovarian cancer cells by inhibiting their expression using the antisense (AS) oligodeoxynucleotide (ODN) technology. Exposure of OV-MZ 6 cells to 10 microM phosphorothioate (PS)-derivatized AS-ODN directed to Rel A led to a maximal 50% decrease of uPA antigen in cell lysates and a 70% reduction in cell cultures supernatants accompanied by a significant transient decline in uPA mRNA levels. Antisense-PS-ODN directed to NF-kappa B1 (p50) or c-rel had no effect on uPA protein expression. AS-PS-ODN directed to Rel A also affected the proteolytic capacity of OV-MZ-6 cells reflected by an approximately 70% decrease in the fibrinolytic capacity of the cells within 24 h compared to untreated controls. AS-PS-ODN directed to I kappa B alpha expression increased uPA in cell culture supernatants up to 50%. uPA receptor (uPAR) production and synthesis of plasminogen activator inhibitor type-1 (PAI-1) were not altered by either AS-PS ODN applied. Western blot and gel retardation analyses revealed constitutive expression of Rel-related proteins in nuclear protein extracts of OV-MZ-6 cells. Thus these proteins seem to be implicated in uPA regulation and may thereby contribute to tumor spread and metastasis. PMID- 7479033 TI - Cleavage of double-stranded DNA by 'metalloporphyrin-linker-oligonucleotide' molecules: influence of the linker. AB - Manganese porphyrin-linker-triple-helix-forming oligonucleotide molecules were prepared and their ability to cleave in vitro a double-stranded DNA target present in the HIV-1 genome was studied. The nature of the linker is a determining factor of the cleavage efficiency. Cleavage yields as high as 80% were observed when the linker was a spermine residue and in the absence of a large excess of free spermine known to stabilize triplex structures. The hydrophobic nature of aliphatic diamine linker modified the cleaver-DNA interactions and reduced the efficiency of DNA cleavage. PMID- 7479034 TI - The orientation of binding of human immunodeficiency virus reverse transcriptase on nucleic acid hybrids. AB - The binding of HIV reverse transcriptase (RT) to heteroduplexes was examined using a substrate consisting of a 42 nt chimeric nucleic acid composed. (5'-->3') of 23 nt of RNA and 19 of DNA. This chimera was hybridized to an internal region of a relatively long complementary DNA or RNA. When the chimera was bound to DNA and conditions limiting cleavage to a single binding event between the enzyme and substrate were employed initial RNase H-directed cleavages occurred 19-21 nt from the chimera 5'-terminus. A 42 nt strand identical in sequence to the chimera and composed of only RNA was cleaved at the same locations. Reducing the length of the DNA portion of the chimera from 19 to 7 nt did not alter the cleavage positions, suggesting that cleavage was not coordinated by the DNA 3'-terminus. Under the same conditions cleavage was not detected when the chimera was bound to RNA. In contrast, addition of dNTPs to the DNA 3'-terminus of the chimera occurred only when the chimera was bound to RNA. The results support preferable binding of RT to RNA-DNA versus DNA-DNA hybrid regions and a model in which the orientation of binding to heteroduplexes is 5'-->3' (relative to the RNA strand), polymerase to RNase H active site, with sites associated with the DNA and RNA strand respectively. PMID- 7479036 TI - Effects of site-specific substitution of 5-fluorouridine on the stabilities of duplex DNA and RNA. AB - The effects of 5-fluorouridine (FUrd) and 5-fluorodeoxyuridine (FdUrd) substitution on the stabilities of duplex RNA and DNA have been studied to determine how FUrd substitution in nucleic acids may alter the efficiency of biochemical processes that require complementary base pairing for molecular recognition. The parent sequence, 5'-GCGAAUUCGC, contains two non-equivalent uridines. Eight oligonucleotides (four RNA and four DNA) were prepared with either zero, one or two Urd substituted by FUrd. The stability of each self complementary duplex was determined by measuring the absorbance at 260 nm as a function of temperature. Tm values were calculated from the first derivative of the absorbance versus temperature profiles and values for delta H0 and delta S0 were calculated from the concentration dependence of the Tm. Individual absorbance versus temperature curves were also analyzed by a parametric approach to calculate thermodynamic parameters for the duplex to single-stranded transition. Analysis of the thermodynamic parameters for each oligonucleotide revealed that FUrd substitution had sequence-dependent effects in both A-form RNA and B-form DNA duplexes. Conservation of helix geometry in FUrd-substituted duplexes was determined by CD spectroscopy. FUrd substitution at a single site in RNA stabilized the duplex (delta delta G37 = 0.8 kcal/mol), largely due to more favorable stacking interactions. FdUrd substitution at a single site in DNA destabilized the duplex (delta delta G37 = 0.3 kcal/mol) as a consequence of less favorable stacking interactions. All duplexes melt via single cooperative transitions. PMID- 7479035 TI - Distinct requirements for primary sequence in the 5'- and 3'-part of a bulge in the hepatitis B virus RNA encapsidation signal revealed by a combined in vivo selection/in vitro amplification system. AB - Hepatitis B virus (HBV) is a small DNA virus that replicates by reverse transcription of a terminally redundant RNA, the pregenome. Specific packaging of this transcript into viral capsids is mediated by interaction of the reverse transcriptase, P protein, with the 5'-proximal encapsidation signal epsilon, epsilon-function is correlated with the formation of a hairpin structure containing a bulge and a loop, each consisting of 6 nt. To analyse the importance of primary sequence in these regions, we have combined selection of encapsidation competent individuals from pools of randomized epsilon-sequences in transfected cells with in vitro amplification, thus bypassing the current experimental limitations of the HBV system. While no alterations of the authentic loop sequence were detectable, many different sequences were tolerated in the 3'-part of the bulge. However, at the two 5'-proximal bulge positions the wt sequence was strongly selected for, indicating that for RNA packaging close contacts between protein and the 5'- but not the 3'-part of the bulge are important. Such a bipartite organisation provides a structural basis for the recently demonstrated special role of the 3'-part of the bulge as template for the first nucleotides of (-)-strand DNA in HBV reverse transcription. PMID- 7479038 TI - A comparison of the in vitro activity of DNA-armed and all-RNA hammerhead ribozymes. AB - Hammerhead ribozymes targeted against two unrelated RNA substrates have been prepared. For each substrate, four ribozymes, differing in their hybridising arm length and composition (DNA or RNA), have been synthesised and kinetically characterised. The presence of DNA in the hybridising arms had little effect on the overall cleavage rate when the cleavage step was rate determining. Shortening each of the hybridising arms of ribozymes from 10 to 6 nucleotides generally resulted in modest changes in rate constants for cleavage of the same 13mer substrate. In one case the presence of long RNA hybridising arms significantly impeded the cleavage reaction. Cleavage rates displayed first order dependence on hydroxide ion concentration at low pHs. At higher pH, some ribozymes deviated from this first order dependence because of a change in the rate-determining step, possibly due to a requirement for a conformation change in the ribozyme substrate complex prior to cleavage. Ribozyme cleavage was strongly dependent on temperature in the range 5-45 degrees C, with an activation energy for the reaction of approximately 60 kJ mol-1. The ribozymes displayed biphasic dependence on magnesium ion concentration; evidence of strong apparent binding (Kd approximately 10 mM) as well as a looser interaction was observed for all ribozymes. PMID- 7479037 TI - In vitro activity of minimised hammerhead ribozymes. AB - A number of minimised hammerhead ribozymes (minizymes) which lack stem II have been kinetically characterised. These minizymes display optimal cleavage activity at temperatures around 37 degrees C. The cleavage reactions of the minizymes are first order in hydroxide ion concentration up to around pH 9.3 above which the cleavage rate constants decline rapidly. The reactions show a biphasic dependence on magnesium-ion concentration; one of the interactions has an apparent dissociation constant of around 20 mM while the other appears to be very weak, showing no sign of saturation at 200 mM MgCl2. The minizymes are significantly less active than comparable, full-size ribozymes when cleaving short substrates. However, at a particular site in a transcribed TAT gene from HIV-1, minizymes are more effective than ribozymes. PMID- 7479039 TI - Mutational analysis of the att DNA-binding domain of phage Mu transposase. AB - The transposase (A protein) of phage Mu encodes binding to two families of DNA sites, att sites located at the Mu ends and enhancer sites located internally. Separate subdomains in the N-terminal domain I of Mu A protein are known to be involved in recognition of the att and enhancer sites. We have delineated an approximately 135 aa region within domain I beta gamma that specifies binding to Mu att sites. This peptide was overexpressed and its properties compared with that of the larger domain I beta gamma as well as the intact Mu A protein. Extensive mutagenesis of residues around a putative helix-turn-helix DNA-binding motif within the I beta domain identified several mutants defective in DNA transposition in vivo. Of these, Mu A(K157Q) was completely defective in att DNA binding. Mu A(F131S) and Mu A(R146N) had a lower affinity for att DNA and low levels of transposition in vitro. Our results indicate that residues in the gamma region are required for activity and that residues outside the beta gamma region must also influence discrimination between the multiple att sites. PMID- 7479040 TI - Mutation detection using immobilized mismatch binding protein (MutS). AB - An accurate and highly sensitive mutation detection assay has been developed. The assay is based on the detection of mispaired and unpaired bases by immobilized mismatch binding protein (Escherichia coli MutS). The assay can detect most mismatches and all single base substitution mutations, as well as small addition or deletion mutations. The assay is simple to use and does not require the use of either radioactivity or gel electrophoresis. PMID- 7479041 TI - Phosphoryl migration during the chemical synthesis of RNA. AB - By the use of high sensitivity assay systems, we have measured the occurrence of strand scission and phosphoryl migration that accompany the deblocking of chemically synthesized oligoribonucleotides. Substantial phosphoryl migration was observed for both enzymatically derived poly(uridylic acid) and synthetic uridine oligoribonucleotides 2'-O-protected with the 1-(2-fluorophenyl)-4 methoxypiperidin-4-yl (Fpmp) group, when these species were subjected to the acidic conditions suggested for Fpmp deprotection. Strand scission occurred in parallel and could be demonstrated readily by 5'-32P end labeling, but not by 3' 32P end labeling, of the acid-treated oligoribonucleotides. Increasing the pH of the deprotection solution and decreasing the temperature at which the deprotection was accomplished diminished both phosphoryl migration and strand scission. A mechanism that can rationalize these results is discussed. PMID- 7479043 TI - The effects of sequence context on base dynamics at TpA steps in DNA studied by NMR. AB - Base dynamics, heretofore observed only at TpA steps in DNA, were investigated as a function of sequence context by NMR spectroscopy. The large amplitude conformational dynamics have been previously observed in TnAn segments where n > or = 2. In order to determine whether the dynamic characteristics occur in more general sequence contexts, we examined four self-complementary DNA sequences, [d(CTTTA-NATNTAAAG)2] (where N = A, C, T, G and N = complement of N). The anomalous broadening of the TpA adenine H2 resonance which is indicative of large amplitude base motion was observed in all nine unique four nucleotide contexts. Furthermore, all the adenine H2 resonances experienced a linewidth maximum as a function of temperature, which is a characteristic of the dynamic process. Interestingly, the temperature of the linewidth maximum varied with sequence indicating that the thermodynamics of TpA base dynamics are also sequence dependent. In one example, neither a T preceding nor an A trailing the TpA step was required for base dynamics. These results show that base dynamics, heretofore observed in only a few isolated sequences, occurs at all TpA steps which are either preceded or followed by a thymine or adenine, respectively, and may be characteristic of all TpA steps in DNA notwithstanding sequence context. PMID- 7479042 TI - Photooxidation of d(TpG) by riboflavin and methylene blue. Isolation and characterization of thymidylyl-(3',5')-2-amino-5-[(2-deoxy-beta-D- erythro pentofuranosyl)amino]-4H-imidazol-4-one and its primary decomposition product thymidylyl-(3',5')-2,2-diamino-4-[(2-deoxy-beta-D- erythro-pentofuranosyl)amino] 5(2H)-oxazolone. AB - The major initial product of riboflavin- and methylene blue-mediated photosensitization of 2'-deoxyguanosine (dG) in oxygen-saturated aqueous solution has previously been identified as 2-amino-5-[(2-deoxy-beta-D-erythro pentofuranosyl)amino] 4H-imidazol-4-one (dlz). At room temperature in aqueous solution dlz decomposes quantitatively to 2,2-diamino-4-[(2-deoxy-beta-D-erythro- pentofuranosyl)amino]-5(2H)-oxazolone (dZ). The data presented here show that the same guanine photooxidation products are generated following riboflavin- and methylene blue-mediated photosensitization of thymidylyl-(3',5')-2' deoxyguanosine [d(TpG)]. As observed for the monomers, the initial product, thymidylyl-(3',5')-2-amino-5-[(2-deoxy- beta-D-erythro-pentofuranosyl)amino]-4H imidazol-4-one [d(Tplz)], decomposes in aqueous solution at room temperature to thymidylyl-(3',5')-2,2-diamino-4- [(2-deoxy-beta-D-erythro-pentofuranosyl)amino] 5(2H)-oxazolone [d(TpZ)]. Both modified dinucleoside monophosphates have been isolated by HPLC and characterized by proton NMR spectrometry, fast atom bombardment mass spectrometry, chemical analyses and enzymatic digestions. Among the chemical and enzymatic properties of these modified dinucleoside monophosphates are: (i) d(Tplz) and d(TpZ) are alkali-labile; (ii) d(Tplz) reacts with methoxyamine, while d(TpZ) is unreactive; (iii) d(Tplz) is digested by snake venom phosphodiesterase, while d(TpZ) is unaffected; (iv) relative to d(TpG), d(TpZ) and d(Tplz) are slowly digested by spleen phosphodiesterase; (v) d(Tplz) and d(TpZ) can be 5'-phosphorylated by T4 polynucleotide kinase. The first observation suggests that dlz and dZ may be responsible for some of the strand breaks detected following hot piperidine treatment of DNA exposed to photosensitizers. PMID- 7479044 TI - Structural basis for the RNA binding selectivity of oligonucleotide analogues containing alkylsulfide internucleoside linkages and 2'-substituted 3' deoxyribonucleosides. AB - In this report we describe the synthesis of oligonucleotides containing sulfide linked dinucleoside units, namely rT(2'OH)sdT, rT(2'OMe)sdT, dTsrU(2'OMe) and dT(2'OMe)srU(2'OMe). We also describe the interactions of such oligomers with complementary DNA and RNA targets, and provide the structural basis for their remarkable RNA binding selectivity. In all cases, the Tm values of the S/P chimera duplexes were lower than those of the corresponding unmodified duplexes. We attribute this to steric interactions between the 5'sulfur and the atoms of the nearby base/sugar residues. The 2'-substituents (i.e., 2'OH or 2'OMe) vicinal to the alkylsulfide internucleoside linkage significantly perturb the structure and stability of the duplexes formed with DNA, and more so than with RNA. The introduction of three rT(2'OH)sdTp (or rT(2'OMe)sdTp) units into an oligodeoxynucleotide sequence was sufficient to abolish binding to complementary DNA but not RNA. The same three substitutions with dTsrU(2'OMe)p and dT(2'OMe)srU(2'OMe)p did not abolish binding to DNA but the resulting complexes had poor thermal stability. The RNA-binding 'selectivity' exhibited by these oligomers is attributed to the tendency of the 2'-substituted (branched) furanoses to adopt the C3'-endo pucker, a conformation that is inconsistent with the B-form structure of helical DNA. The preference of these sugars to exist often exclusively in the C3'-endo form is attributed to stereoelectronic effects, namely gauche and anomeric effects. Our findings support the hypothesis that nucleoside analogues puckered exclusively in the C3'-endo form may result in them being especially good binders of targeted mRNA [S.H. Kawai (1991), Ph.D. Thesis, McGill University; Kawasaki et al. (1993) J. Med. Chem. 36, 831-841]. PMID- 7479045 TI - The ribonuclease activity of nucleolar protein B23. AB - Protein B23 is an abundant nucleolar protein and putative ribosome assembly factor. The protein was analyzed for ribonuclease activity using RNA-embedded gels and perchloric acid precipitation assays. Three purified bacterially expressed forms of the protein, B23.1, B23.2 and an N-terminal polyhistidine tagged B23.1 as well as the natural protein were found to have ribonuclease activity. However, the specific activity of recombinant B23.1 was approximately 5 fold greater than that of recombinant B23.2. The activity was insensitive to human placental ribonuclease inhibitor, but was inhibited by calf thymus DNA in a dose dependent manner. The enzyme exhibited activity over a broad range of pH with an apparent optimum at pH 7.5. The activity was stimulated by but not dependent on the presence of low concentrations of Ca2+, Mg2+ or NaCl. The Ca2+ effect was saturable and only stimulatory in nature. In contrast, Mg2+ and NaCl exhibited optimal concentrations for stimulation and both inhibited the ribonuclease at concentrations above these optima. These data suggest that protein B23 has intrinsic ribonuclease activity. The location of protein B23 in subcompartments of the nucleolus that contain preribosomal RNA suggests that its ribonuclease activity plays a role in the processing of preribosomal RNA. PMID- 7479046 TI - Sequences 5' of the first upstream open reading frame in GCN4 mRNA are required for efficient translational reinitiation. AB - Translation of yeast GCN4 mRNA occurs by a reinitiation mechanism that is modulated by amino acid levels in the cell. Ribosomes which translate the first of four upstream open reading frames (uORFs) in the mRNA leader resume scanning and can reinitiate downstream. Under non-starvation conditions reinitiation occurs at one of the remaining three uORFs and GCN4 is repressed. Under starvation conditions, in contrast, ribosomes bypass the uORFs and reinitiate at GCN4 instead. The high frequency of reinitiation following uORF1 translation depends on an adequate distance to the next start codon and particular sequences surrounding the uORF1 stop codon. We present evidence that sequences 5' to uORF1 also strongly enhance reinitiation. First, reinitiation was severely inhibited when uORF1 was transplanted into the position of uORF4, even though the native sequence environment of the uORF1 stop codon was maintained, and this effect could not be accounted for by the decreased uORF1-GCN4 spacing. Second, insertions and deletions in the leader preceding uORF1 greatly reduced reinitiation at GCN4. Sequences 5' to uORF1 may influence the probability of ribosome release following peptide termination at uORF1. Alternatively, they may facilitate rebinding of an initiation factor required for reinitiation prior to resumption of the scanning process. PMID- 7479047 TI - Synthesis and biological activities of a phosphorodithioate analog of 2',5' oligoadenylate. AB - To enhance the resistance of 2-5A (pppA2'p5'A2'p5'A) to degradation by exo- and endonucleases, a phosphorodithioate analog was synthesized using a solid-phase phosphite triester approach with N6-benzoyl-5'-O-dimethoxytrityl-3'-O-t butyldimethylsilyladenosine 2'-[S-(beta-thiobenzoylethyl) pyrrolidinophosphorothioamidit e]. 5'-Monophosphorylation was accomplished with 2 [2-(4,4'-dimethoxytrityloxy)-ethylsulfonyl]ethyl-(2-cyanoe thyl)-(N,N- diisopropyl)-phosphoramidite. The resulting product, p5'A2'(s2p)- 5'A2'(s2p)5'A, was approximately 10-fold less effective as an activator of purified human recombinant 2-5A-dependent RNase than was 2-5A itself. This loss of activation ability was related directly to the loss of binding ability of the phosphorodiothioate analog. As predicted, p5'A2'(s2p)5'A2' (s2p)5'A was stable to snake venom phosphodiesterase and the nucleolytic activities of both human lymphoblastoid CEM cell extracts and human serum, under conditions that led to facile degradation of parent 2-5A. This nuclease stability permitted the observation of the CEM cell extracts and human serum phosphatase activity which led to 5'-dephosphorylation of p5'A2'(s2p)5'A2'(s2p)5'A. PMID- 7479049 TI - Rapid DNA preparation for 2D gel analysis of replication intermediates. PMID- 7479048 TI - Large scale isolation of expression vector cassette by magnetic triple helix affinity capture. PMID- 7479050 TI - Protection of megabase DNA from shearing. PMID- 7479051 TI - Hydroxyquinoline overcomes PCR inhibition by UV-damaged mineral oil. PMID- 7479052 TI - A lacZ-hygromycin fusion gene and its use in a gene trap vector for marking embryonic stem cells. PMID- 7479053 TI - Construction of specific cosmids from YACs by homologous recombination in yeast. PMID- 7479055 TI - Functional expression of a single-chain anti-progesterone antibody fragment in the cytoplasm of a mutant Escherichia coli. PMID- 7479054 TI - Identification of a differentially-expressed message associated with colon cancer liver metastasis using an improved method of differential display. PMID- 7479056 TI - Insertion of unique sites into YAC arms for rapid physical analysis following YAC transfer into mammalian cells. PMID- 7479057 TI - A rapid procedure for the compositional analysis of yeast artificial chromosomes. PMID- 7479058 TI - Aberrant pre-mRNA maturation is caused by LINE insertions into introns of the white gene of Drosophila melanogaster. AB - Insertional mutagenesis screens have provided thousands of mutant alleles for analysing genes of varied functions in Drosophila melanogaster. We here document mechanisms of insertional mutagenesis by a LINE element, the I factor, by determining the molecular structure of RNAs produced from two alleles of the white gene of D.melanogaster, wIR1 and wIR6. These alleles result from insertion of the I factor into introns of the gene. We show that sequences present within the element direct aberrant splicing and termination events. When the I factor is inserted within the white first intron it may lead to the use of a cryptic 3' splice site which does not contain the dinucleotide AG. This splicing gives rise to a chimeric messenger RNA whose synthesis is controlled differently in tissues where the mutated gene is expressed. When the I factor is inserted within the white last intron it induces synthesis of truncated mRNAs. These results provide, for the first time, mechanisms for I factor insertional mutagenesis. They are discussed in the more general context of RNA processing in Drosophila and the evolution of eukaryotic gene introns. PMID- 7479059 TI - Species- and tissue-specific expression of the C-terminal alternatively spliced form of the tumor suppressor p53. AB - Alternative splicing of the p53 transcript which so far has been demonstrated only in the murine system has been proposed as a general regulatory mechanism for the generation of functionally different p53 proteins. We analyzed by RT-PCR the pattern of p53 mRNAs within the region spanning exons 10 and 11 of the p53 gene in 13 different tissues from two independent mouse strains, in 10 different rat tissues and in six different human tissues. PCR products of the expected sizes, corresponding to the normally spliced and the alternatively spliced p53 mRNAs, were detected in mice. Alternatively spliced mRNA was found at approximately 25 20% the level of the normally spliced p53 mRNA in most tissues analyzed. In spleen and kidney the proportion of alternatively spliced p53 mRNA was much lower. Surprisingly, examination of p53 mRNAs isolated from 10 different rat tissues and six human tissues within the same region of the p53 gene showed only products of normal size. Although a potential homologous alternative 3' splice site within intron 10 of the human p53 gene is present in the genomic sequence of human p53, the expected corresponding alternatively spliced p53 mRNA was undetectable. These findings imply that the generation of functionally different forms of p53 by alternative splicing of p53 transcripts is a species-specific event, possibly indicating species-specific mechanisms for regulating p53 activities. PMID- 7479060 TI - New efficient sulfurizing reagents for the preparation of oligodeoxyribonucleotide phosphorothioate analogues. AB - A set of new sulfurizing agents representing disulfides of arylsulfonic acids has been developed for the automated synthesis of phosphorothioate oligonucleotide analogues via the phosphoramidite method. These reagents, such as bis(benzenesulfonyl)disulfide, bis(p-toluenesulfonyl)disulfide, bis(p methoxybenzensulfonyl)disulfide, and bis (p-chlorobenzenesulfonyl) disulfide, are easily prepared crystalline solid compounds. They are relatively inexpensive, easy to handle, and efficiently convert internucleotide cyanoethyl phosphite to the phosphorothioate triester within 1-2 min. The efficiency of phosphorothioate oligonucleotide synthesis with the use of these reagents is comparable to that of phosphodiester oligonucleotides. PMID- 7479061 TI - Isolation of genomic and cDNA clones encoding bovine poly(A) binding protein II. AB - cDNA clones for bovine poly(A) binding protein II (PAB II) were isolated. Their sequence predicts a protein of 32.8 kDa, revising earlier estimates of molecular mass. The protein contains one putative RNA-binding domain of the RNP type, an acidic N-terminal and a basic C-terminal domain. Analyses of authentic PAB II were in good agreement with all predictions from the cDNA sequence except that a number of arginine residues appeared to be post-translationally modified. Poly(A) binding protein II expressed in Escherichia coli was active in poly(A) binding and reconstitution of processive polyadenylation, including poly(A) tail length control. The cDNA clones showed a number of potential PAB II binding sites in the 3' untranslated sequence. Bovine poly(A)+RNA contained two mRNAs hybridizing to a PAB II-specific probe. Analysis of a genomic clone revealed six introns in the coding sequence. The revised molecular mass led to a demonstration of PAB II oligomer formation and a reinterpretation of earlier data concerning the protein's binding to poly(A). PMID- 7479062 TI - Triplex formation with alpha anomers of purine-rich and pyrimidine-rich oligodeoxynucleotides. AB - Nuclease-resistant alpha anomers of pyrimidine-rich CT- and purine-rich GA- and GT-containing oligonucleotides were investigated for their triplex-forming potential and compared with their corresponding nuclease-sensitive beta anomers. Both 23mer CT-alpha and 23mer CT-beta had quite similar triplex binding affinities. Synthetic 23mer GT-alpha oligonucleotides were capable of triplex formation with binding affinities slightly lower than corresponding 23mer GT-beta oligonucleotides. The orientation of third strand GT-alpha binding was parallel to the purine strand of the duplex DNA target, whereas the orientation of third strand GT-beta binding was found to be antiparallel. Triplex formation with both GT oligonucleotides showed the typical dependence on magnesium and temperature. In contrast, 23mer GA-alpha oligonucleotides did not support triplex formation in either orientation under a variety of experimental conditions, whereas the corresponding 23mer GA-beta oligonucleotides demonstrated strong triplex formation in the antiparallel orientation. GA-alpha oligonucleotides covalently conjugated to acridine were similarly unable to demonstrate triplex formation. GA alpha oligonucleotides, in contrast to GT-alpha oligonucleotides, were capable of self-association, detectable by gel retardation and UV spectroscopy, but competing self-association could not fully account for the lack of triplex formation. Thus for in vivo triplex gene regulation strategies using GT oligonucleotides the non-natural alpha anomer may be a feasible alternative to the natural beta anomer, allowing for a comparable degree of triplex formation without rapid cellular degradation. However, alpha anomeric inversion does not appear to be a feasible alternative in applications involving GA oligonucleotides. PMID- 7479063 TI - Cell cycle-dependent regulation of RNA polymerase II basal transcription activity. AB - Regulation of transcription by RNA polymerase II (pol II) in eukaryotic cells requires both basal and regulatory transcription factors. In this report we have investigated in vitro pol II basal transcription activity during the cell cycle by using nuclear extracts from synchronized HeLa cells. It is shown that pol II basal transcription activity is low in the S and G2 phases and high in early G1 phase and TFIID is the rate limiting component of pol II basal transcription activity during the cell cycle. Further analyses reveal that TFIID exists as a less active form in the S and G2 phases and nuclear extracts from S and G2 phase cells contain a heat-sensitive repressor(s) of TATA box binding protein (TBP). These results suggest that pol II basal transcription activity is regulated by a qualitative change in the TFIID complex, which could involve repression of TBP, during the cell cycle. PMID- 7479064 TI - The purine-rich trinucleotide repeat sequences d(CAG)15 and d(GAC)15 form hairpins. AB - The structures of single-stranded (ss) oligonucleotides containing (CAG)15 [ss(CAG)15] or (GAC)15 [ss(GAC)15] were examined. At 10 degrees C, the electrophoretic mobilites of the two DNAs were similar to ss(CTG)15, a DNA that forms a hairpin containing base paired and/or stacked thymines. At 37 degrees C in 50 mM NaCl, single-strand-specific P1 nuclease cleaved the G33-G36 phosphodiesters of ss(GAC)15, and the G32-A34, G35-C36 phosphodiesters of ss(CAG)15 (where the loop apex of both DNAs = A34). Electrophoretic mobility melting profiles indicated that the melting temperature (Tm) of ss(CAG)15 in low (approximately 1 mM Na+) ionic strength was 38 degrees C. In contrast, the Tm of ss(GAC)15 was 49 degrees C, a value similar to the Tm of ss(CTG)15. These results provide evidence that ss(GAC)15 and ss(CAG)15 form similar, but distinguishable hairpin structures. PMID- 7479065 TI - NMR analysis of the trans-activation response (TAR) RNA element of equine infectious anemia virus. AB - Transcription of lentiviral DNA in the host cell is regulated by an interaction between the viral TAR RNA stem-loop and the viral Tat protein. Here we present a model of the three-dimensional structure of the TAR RNA stem-loop of the equine infectious anemia virus (EIAV), derived from two- and three-dimensional NMR data. This 25 nucleotide RNA consists of an A-form helical stem capped by two U-G base pairs and a four-nucleotide loop. Two loop cytidines are stacked into the loop interior and likely form a non-Watson-Crick C-C base-pair. The two nucleotides at the top of the loop, U13 and G14, appear to be excluded from the interior of the loop and solvent exposed. It is significant that now for the EIAV TAR-Tat system, three-dimensional structures are now known for both the RNA and protein components. PMID- 7479066 TI - Mutagenic and genotoxic effects of DNA adducts formed by the anticancer drug cis diamminedichloroplatinum(II). AB - The toxicity and mutagenicity of three DNA adducts formed by the anticancer drug cis-diamminedichloroplatinum(II) (cis-DDP or cisplatin) were investigated in Escherichia coli. The adducts studied were cis-[Pt(NH3)2(d(GpG))] (G*G*), cis [Pt(NH3)2(d(ApG))] (A*G*) and cis-[Pt(NH3)2(d(GpTpG))] (G*TG*), which collectively represent approximately 95% of the DNA adducts reported to form when the drug damages DNA. Oligonucleotide 24-mers containing each adduct were positioned at a known site within the viral strand of single stranded M13mp7L2 bacteriophage DNA. Following transfection into E. coli DL7 cells, the genomes containing the G*G*, A*G* and G*TG* adducts had survival levels of 5.2 +/- 1.2, 22 +/- 2.6 and 14 +/- 2.5% respectively, compared to unmodified genomes. Upon SOS induction, the survival of genomes containing the G*G* and A*G* adducts increased to 31 +/- 5.4 and 32 +/- 4.9% respectively. Survival of the genome containing the G*TG* adduct did not increase upon SOS induction. In SOS induced cells, the G*G* and A*G* adducts gave rise predominantly to G-->T and A-->T transversions respectively, targeted to the 5' modified base. In addition, A-->G transitions were detected for the A*G* adduct and low levels of tandem mutations at the 5' modified base as well as the adjacent 5' base were also observed for both adducts. The A*G* adduct was more mutagenic than the G*G* adduct, with a mutation frequency of 6% compared to 1.4% for the latter adduct. No cis-[Pt(NH3)2)2+ intrastrand crosslink-specific mutations were observed for the G*TG* adduct. PMID- 7479067 TI - The expression of biologically active human p53 in Leishmania cells: a novel eukaryotic system to produce recombinant proteins. AB - We have investigated the use of Leishmania cells as a novel eukaryotic expression system for the production of recombinant protein. These cells are easy to maintain, requiring no CO2 incubator or shaker, and can be grown in standard tissue culture media. Leishmania cells can be readily transfected with plasmid DNA by electroporation and transformants selected with antibiotic resistance. Recent studies have shown that it is possible to express foreign genes in Leishmania for the purpose of understanding the biology of this protozoan cell. In the present study we report the use of this system as a means of producing a biologically functional human p53 protein. The conformation of the p53 protein is critical for its ability to bind specific DNA sequences. It is demonstrated that Leishmania-synthesized human p53 is phosphorylated and can bind specifically to its enhancer DNA sequence. These data demonstrate that Leishmania may represent a simple eukaryotic expression system for the production of biologically active recombinant proteins. PMID- 7479068 TI - Members of a family of proteins (the RD family) detected by a U1 70K monoclonal antibody are present in spliceosomal complexes. AB - We have characterized a monoclonal antibody (mAb) to the U1 snRNP component U1 70K. We find that this antibody recognizes several proteins, in addition to U1 70K, in purified spliceosomal complexes and in total HeLa cell nuclear extract preparations. The novel mAb U1 70K antigens can also be specifically immunoprecipitated by the antibody. Similarly to U1 70K, many of the mAb U1 70K antigens can be phosphorylated by a co-purifying kinase activity. The epitope recognized by mAb U1 70K was previously shown to be a repeating arginine/aspartate (RD) dipeptide. Thus we have designated the novel mAb U1 70K antigens the RD family. Comparison of mAb U1 70K with a recently characterized antibody, mAb 16H3, whose epitope is a repeating R/D or R/E motif, showed that a large subset of the antigens are common. In contrast, most of the mAb U1 70K antigens are distinct from the proteins detected by mAb 104, an antibody to the SR family of splicing factors. PMID- 7479069 TI - Mutational analysis of a conserved motif of Agrobacterium tumefaciens VirD2. AB - The VirD2 polypeptide from Agrobacterium tumefaciens, in the presence of VirD1, introduces a site- and strand-specific nick at the T-DNA borders. A similar reaction at the origin of transfer (oriT) of plasmids is essential for plasmid transfer by bacterial conjugation. A comparison of protein sequences of VirD2 and its functional homologs in bacterial conjugation and in rolling circle replication revealed that they share a conserved 14 residue segment, HxDxxx(P/u)HuHuuux [residues 126-139 of VirD2; Ilyina, T.V. and Koonin, E.V. (1992) Nucleic Acids Res. 20, 3279-3285]. A mutational approach was used to test the role of these residues in the endonuclease activity of VirD2. The results demonstrated that the two invariant histidine residues (H133 and H135) are essential for activity. Mutations at three sites, histidine 126, aspartic acid 128 and aspartic acid 130, that are conserved in a subfamily of the plasmid mobilization proteins, led to the loss of VirD2 activity. Aspartic acid at position 130, could be substituted with glutamic acid and to a much lesser extent, with tyrosine. In contrast, another conserved residue, asparagine 139, tolerated many different amino acid substitutions. The non-conserved residues, arginine 129, proline 132 and leucine 134, were also found to be important for function. Isolation of null mutations that map throughout this conserved domain confirm the hypothesis that this region is essential for function. PMID- 7479070 TI - An oligodeoxyribonucleotide that supports catalytic activity in the hammerhead ribozyme domain. AB - A study of the activity of deoxyribonucleotide-substituted analogs of the hammerhead domain of RNA catalysis has led to the design of a 14mer oligomer composed entirely of deoxyribonucleotides that promotes the cleavage of an RNA substrate. Characterization of this reaction with sequence variants and mixed DNA/RNA oligomers shows that, although the all-deoxyribonucleotide oligomer is less efficient in catalysis, the DNA/substrate complex shares many of the properties of the all-RNA hammerhead domain such as multiple turnover kinetics and dependence on Mg2+ concentration. On the other hand, the values of kinetic parameters distinguish the DNA oligomer from the all-RNA oligomer. In addition, an analog of the oligomer having a single ribonucleotide in a strongly conserved position of the hammerhead domain is associated with more efficient catalysis than the all-RNA oligomer. PMID- 7479072 TI - Selection of aminoacyl-tRNAs at sense codons: the size of the tRNA variable loop determines whether the immediate 3' nucleotide to the codon has a context effect. AB - Codon context can affect translational efficiency by several molecular mechanisms. The base stacking interactions between a codon-anticodon complex and the neighboring nucleotide immediately 3' can facilitate translation by amber suppressors and the tRNA structure is also known to modulate the sensitivity to context. In this study the relative rates of aminoacyl-tRNA selection were measured at four sense codons (UGG, CUC, UUC and UCA), in all four 3' nucleotide contexts, through direct competition with a programmed frameshift at a site derived from the release factor 2 gene. Two codons (UGG and UUC) are read by tRNAs with small variable regions and their rates of aminoacyl-tRNA selection correlated with the potential base stacking strength of the 3' neighboring nucleotide. The other two codons (CUC and UCA) are read by tRNAs with large variable regions and the rate of selection of the aminoacyl-tRNAs in these cases varied little among the four contexts. Re-examination of published data on amber suppression also revealed an inverse correlation between context sensitivity and the size of the variable region. Collectively the data suggest that a large variable loop in a tRNA decreases the influence of the 3' context on tRNA selection, probably by strengthening tRNA-ribosomal interactions. PMID- 7479073 TI - Interactions of calf thymus DNA polymerase alpha with primer/templates. AB - The interactions of calf thymus DNA polymerase alpha (pol alpha) with primer/templates were examined. Simply changing the primer from DNA to RNA had little effect on primer/template binding or dNTP polymerization (Km, Vmax and processivity). Surprisingly, however, adding a 5'-triphosphate to the primer greatly changed its interactions with pol alpha (binding, Vmax and Km and processivity). While changing the primer from DNA to RNA greatly altered the abilit of pol alpha to discriminate against nucleotide analogs, it did not compromise the ability of pol alpha to discriminate against non-cognate dNTPs. Thus the nature of the primer appears to affect 'sugar fidelity', without altering 'base fidelity'. DNase protection assays showed that pol alpha strongly protected 9 nt of the primer strand, 13 nt of the duplex template strand and 14 nt of the single-stranded template from hydrolysis by DNase I and weakly protected several bases outside this core region. This large DNA binding domain may account for the ability of a 5'-triphosphate on RNA primers to alter the catalytic properties of pol alpha. PMID- 7479071 TI - A compilation of composite regulatory elements affecting gene transcription in vertebrates. AB - Over the past years, evidence has been accumulating for a fundamental role of protein-protein interactions between transcription factors in gene-specific transcription regulation. Many of these interactions run within composite elements containing binding sites for several factors. We have selected 101 composite regulatory elements identified experimentally in the regulatory regions of 64 genes of vertebrates and of their viruses and briefly described them in a compilation. Of these, 82 composite elements are of the synergistic type and 19 of the antagonistic type. Within the synergistic type composite elements, transcription factors bind to the corresponding sites simultaneously, thus cooperatively activating transcription. The factors, binding to their target sites within antagonistic type composite elements, produce opposing effects on transcription. The nucleotide sequence and localization in the genes, the names and brief description of transcription factors, are provided for each composite element, including a representation of experimental data on its functioning. Most of the composite elements (3/4) fall between -250 bp and the transcription start site. The distance between the binding sites within the composite elements described varies from complete overlapping to 80 bp. The compilation of composite elements is presented in the database COMPEL which is electronically accessible by anonymous ftp via internet. PMID- 7479074 TI - NMR characterisation of a triple stranded complex formed by homo-purine and homo pyrimidine DNA strands at 1:1 molar ratio and acidic pH. AB - Homo-purine (d-TGAGGAAAGAAGGT) and homo-pyrimidine (d-CTCCTTTCTTCC) oligomers have been designed such that they are complementary in parallel orientation. When mixed in a 1:1 molar ratio, the system adopts an antiparallel duplex at neutral pH with three mismatched base pairs. On lowering the pH below 5.5, a new complex is formed. The NMR results show the coexistence of a intermolecular pyrimidine.purine:pyrimidine DNA triplex and a single stranded oligopurine at this pH. The triplex is stabilized by five T.A:T, four C+.G:C and two mismatched triads, namely, C+.G-T and T.A-C. This triplex is further stabilized by a Hoogsteen C+.G base-pair on one end. Temperature dependence of the imino proton resonances reveals that the triplex dissociates directly into single strands around 55 degrees C, without duplex intermediates. Parallel duplexes are not formed under any of the conditions employed in this study. PMID- 7479075 TI - Marked increase in the number and variety of mitochondrial DNA rearrangements in aging human skeletal muscle. AB - Several reports have shown that individual mitochondrial DNA (mtDNA) deletions accumulate with age. However, the overall extent of somatic mtDNA damage with age remains unclear. We have utilized full-length PCR to concurrently screen for multiple mtDNA rearrangements in total DNA extracted from skeletal muscle derived from physiologically normal individuals (n = 35). This revealed that both the number and variety of mtDNA rearrangements increases dramatically between young and old individuals (P < 0.0001). We further examined the mtDNA from both the younger and older subjects by Southern blot analysis and observed an age-related increase in mtDNA(s) comparable in size to mtDNA products unique to patients with known mtDNA deletions. These data imply that a wide spectrum of mtDNA rearrangements accumulate in old individuals, which correlates with the marked age related decrease in OXPHOS capacity observed in post-mitotic tissues. PMID- 7479076 TI - Isolation of YAC insert sequences by representational difference analysis. AB - We present a method for the isolation of YAC insert sequences by representational difference analysis (RDA). To achieve maximal representation of the sequences, the amplicons were generated from a Mbol digestion product. RDA was performed using a 970 kb insert YAC clone. After two rounds of re-association and selective amplification 92% of the difference product represented sequences derived from the YAC insert. Twenty insert-specific sequence-tagged sites were readily defined. The difference product was also successfully used to isolate microsatellite markers, to identify clones from a human PAC library and as a chromosome painting probe in fluorescence in situ hybridization. PMID- 7479077 TI - Stability and functional effectiveness of phosphorothioate modified duplex DNA and synthetic 'mini-genes'. AB - Several gene transfer techniques that employ 'naked DNA' molecules have recently been developed and numerous gene therapy protocols that make use of 'naked-DNA' have been proposed. We studied the possibility of enhancing the stability of 'naked DNA vectors' and thus also gene transfer and expression efficiencies, by constructing phosphorothioate (PS-) double strand DNA molecules and functional transcription units. We first synthesized short PS-double strand DNA molecules by the annealing of two complementary, 35 nt long, oligonucleotides. The accessibility of DNA modifying enzymes to this molecule was significantly decreased: T4-ligase and kinase activity were respectively reduced up to 1/2 and to 1/6, as compared to the normal phosphodiester molecule. Nucleolytic stability was increased either to purified enzymes (DNase I and Bal31) or to incubations in fresh serum, cell culture medium or in muscle protein extract. Phosphorothioate end-capped complete eukaryotic transcription units (obtained by Taq polymerase amplification with PS-primers) were not significantly protected from nucleolytic attack. On the contrary, synthetic transcription units, 'mini genes', obtained by Taq amplification with 1, 2 or 3 PS-dNTP substitutions, were resistant to DNase I and Bal31 nucleolytic activity. Transcription efficiency, driven by the T7 promoter, was 96.5, 95 and 33.5% (respectively with 1, 2 or 3 substitutions), as compared to the normal phosphodiester molecules. PMID- 7479078 TI - COUP-TF II homodimers are formed in preference to heterodimers with RXR alpha or TR beta in intact cells. AB - Chicken ovalbumin upstream promoter-transcription factor (COUP-TF) represses the transcriptional activity of a number of nuclear receptors, including that of retinoid receptors (RAR and RXR) and thyroid hormone receptors (TR). Since COUP TF is capable of binding to DNA in vitro either as a homodimer or as a heterodimer with RXR or TR, it has not been possible to distinguish between competitive DNA binding and heterodimer formation as a mechanism to account for the repression. Using a two-hybrid system we have investigated the dimerisation properties of COUP-TF II in intact cells. In conditions where COUP-TF II homodimers and RXR alpha-RAR alpha heterodimers were formed we were unable to detect the formation of heterodimers between COUP-TF II and RXR alpha. Moreover, we were unable to detect an interaction between COUP-TF II and RXR alpha on DNA. Similarly COUP-TF II homodimers and RXR alpha-TR beta heterodimers are favoured over COUP-TF II-TR beta heterodimers. We conclude that the formation of functionally inactive heterodimers is unlikely to represent a general mechanism by which COUP-TF represses the transcriptional activity of nuclear receptors and favour a model in which repression is mediated by COUP-TF homodimers competing for binding to DNA. PMID- 7479079 TI - Solid-phase synthesis of a nucleopeptide from the linking site of adenovirus-2 nucleoprotein, -Ser(p5'CATCAT)-Gly-Asp-. Convergent versus stepwise strategy. AB - The synthesis of a nucleopeptide with the sequence -Ser(p5'CATCAT)-Gly-Asp- has been undertaken by either convergent or stepwise solid-phase strategies, both of which use base-labile permanent protecting groups. The coupling of phosphitylated protected peptides onto oligonucleotide-resins did not afford the desired nucleopeptide, which was nevertheless obtained after oligonucleotide elongation at the hydroxyl group of the resin-bound peptide and deprotection under mild basic conditions. A preliminary study on the stability of different nucleopeptides to bases is also reported. PMID- 7479080 TI - A comparison of the different DNA binding specificities of the bZip proteins C/EBP and GCN4. AB - The bZip proteins GCN4 and C/EBP differ in their DNA binding specificities: GCN4 binds well to the pseudopalindromic AP1 site 5'-A4T3G2A1C0T1C2'A3'T4'-3' and to the palindromic ATF/CREB sequence 5'-A4T3G2A1-C0*G0'T1'C2'A3'T4'-3'; C/EBP preferentially recognizes the palindromic sequence 5'-A4T3T2G1C0*G0'C1'A2'-A3'T4' 3'. According to the X-ray structures of GCN4-DNA complexes, five residues of the basic region of GCN4 are involved in specific base contacts: asparagine -18, alanine -15, alanine -14, serine -11 and arginine -10 (numbered relative to the start point of the leucine zipper, which we define as +1). In the basic region of C/EBP position -14 is occupied by valine instead of alanine, the other four residues being identical. Here we analyse the role of valine -14 in C/EBP-DNA complex formation. Starting from a C/EBP-GCN4 chimeric bZip peptide which displays C/EBP specificity, we systematically mutated position -14 of its basic region and characterized the DNA binding specificities of the 20 possible different peptides by gel mobility shift assays with various target sites. We present evidence that valine -14 of C/EBP interacts more strongly with thymine 2 than with cytosine 1' of the C/EBP binding site, unlike the corresponding alanine -14 of GCN4, which exclusively contacts thymine 1' of the GCN4 binding sites. PMID- 7479081 TI - Modified curved DNA that could allow local DNA underwinding at the nucleosomal pseudodyad fails to position a nucleosome in vivo. AB - In competitive in vitro reconstitution experiments synthetic DNA composed of tandem repeats of the repetitive sequence (A/T)3NN(G/C)3NN, specifically the 20 bp 'TG sequence' (5'-TCGGTGTTAGAGCCTGTAAC-3'), was reported to associate with the histone octamer with an affinity higher than that of nucleosomally derived DNA. However, at least two groups have independently shown that tandem repeats of the TG sequence do not accommodate a stably positioned nucleosome in vivo. It was suggested that the anisotropic flexibility of the TG sequence, governed by a 10 bp sequence periodicity, is incompatible with the required underwinding of the DNA helix at the nucleosome pseudodyad while maintaining a bending preference that can be accommodated in the remainder of the nucleosome. Here we test this hypothesis directly by studying the in vivo nucleosomal structure of modified TG sequences designed to accommodate underwinding at the pseudodyad. We show that these modifications are not sufficient to allow stable incorporation of the TG sequence repeat into a nucleosome in vivo, but do note invasion from one end of the TG heptamer of a translationally random but rotationally constrained nucleosome. We discuss possible reasons for the absence of nucleosomes from the TG sequence in vivo. PMID- 7479082 TI - 3-D models of the antigenomic ribozyme of the hepatitis delta agent with eight new contacts suggested by sequence analysis of 188 cDNA clones. AB - We mapped 359 mutations at 25 positions in synthetic variants of the antigenomic ribozyme of the hepatitis delta agent by analyzing the sequences of 188 cDNA clones. These data were used to identify three features of the ribozyme: highly conserved nucleotides, positions with restricted nucleotide substitutions and three-dimensional relationships between nucleotides. The distribution of mutations at the 25 positions was as follows: G-11 (the eleventh nucleotide from the cleavage site) was mutated in 56 clones; G-12 in 36; U-15 in 33; C-13 in 26; G-28 in 23; C-27 in 21; C-29 in 19; U-26 in 17; C-18 in 14; A-14 in 13; C-16 in 13; C-19 in 12; U-17 in 11; A-20 in 10; G-42 in 9; G-40 in 7; G-41 in 7; C-24 in 6; U-32 in 6; U-23 in 5; C-25 in 4; C-21 in 3; G-30 in 3; G-31 in 3; C-22 in 1. All clones containing a mutation at C-25 had an A at this position, suggesting that the extra cyclic amino group present in adenine and cytosine may function during the cleavage event. Mutations at certain positions were common in simple clones (containing only one or two mutations), while mutations at other positions were over-represented in more complex clones. Both compensatory base changes and co-mutational frequencies were used to identify eight pairs of nucleotides which may interact with each other: G-11 and C-18, G-12 and C-27, C-13 and G-28, C-21 and U-23/C-24, C-21 and G-30, U-23 and G-31/U-32, C24 and G-30, C-27 and G-42. These pairs, which involve some of the most conserved positions in the molecule, suggest interactions among nucleotides previously depicted in open-loop structures. The newly proposed points of contact between pairs of nucleotides are compatible with both the axehead and pseudoknot secondary structural models and were combined with previously proposed Watson-Crick base paired helices to produce two three dimensional models. In both of these, C-25 and C-76 are placed near the cleavage site. PMID- 7479083 TI - Role of RNA secondary structure of the iron-responsive element in translational regulation of ferritin synthesis. AB - Iron regulates synthesis of the iron storage protein ferritin at the translational level through interaction between a stem-loop structure, the iron responsive element (IRE), located in the 5'-untranslated region (5'-UTR) of ferritin mRNAs, and a protein, the iron regulatory protein (IRP). The role of IRE secondary structure in translational regulation of ferritin synthesis was explored by introducing ferritin constructs containing mutations in the IRE into Rat-2 fibroblasts. Our in vivo studies demonstrate that size and sequence of the loop within the IRE and the distance and/or spatial relationship of this loop to the bulged nucleotide region closest to the loop must be preserved in order to observe iron-dependent translation of ferritin mRNA. In contrast, changes in nucleotide sequence of the upper stem can be introduced without affecting translational regulation in vivo, as long as a stem can be formed. Our in vivo results suggest that only a very small variation in the affinity of interaction of IRP with IRE can be tolerated in order to maintain iron-dependent regulation of translation. PMID- 7479084 TI - Initiation of unidirectional ColE2 DNA replication by a unique priming mechanism. AB - The ColE2 DNA can be replicated in an in vitro system consisting of a crude extract of Escherichia coli cells. DNA synthesis requires a plasmid-coded protein (Rep) and host DNA polymerase I but not host RNA polymerase. Replication starts at a fixed region containing the origin and proceeds unidirectionally. The leading- and lagging-strand DNA fragments synthesized around the origin were identified from early replicative intermediates. The 5' end of the leading-strand DNA fragment was mapped at a unique position in the minimal origin and carried RNA of a few residues. The results suggested that the initiation of the leading strand DNA synthesis does not require the host DnaG protein. Thus the Rep protein itself seems to be a primase. Synthesis of the primer RNA at a fixed site in the origin region on a double-stranded DNA template is a unique property of the ColE2 Rep protein among other known primases. The 3' end of the lagging-strand DNA fragment was mapped at a unique position just at the end of the minimal origin region. Termination of the lagging-strand DNA fragment at that position seems to be the mechanism of the unidirectional replication of ColE2 plasmid. PMID- 7479085 TI - CGG repeats associated with DNA instability and chromosome fragility form structures that block DNA synthesis in vitro. AB - A large increase in the length of a CGG tandem array is associated with a number of triplet expansion diseases, including fragile X syndrome, the most common cause of heritable mental retardation in humans. Expansion results in the appearance of a fragile site on the X chromosome in the region of the CGG array. We show here that CGG repeats readily form a series of barriers to DNA synthesis in vitro. There barriers form only when the (CGG)n strand is used as the template, are K(+)-dependent, template concentration-independent, and involve hydrogen bonding between guanines. Chemical modification experiments suggest these blocks to DNA synthesis result from the formation of a series of intrastrand tetraplexes. A number of lines of evidence suggest that both triplet expansion and chromosome fragility are the result of replication defects. Our data are discussed in the light of such evidence. PMID- 7479087 TI - PCR product with strands of unequal length. PMID- 7479089 TI - Transverse temperature-gradient single-strand conformation polymorphism analysis for temperature optimization of 'Cold'-SSCP mutation detection. PMID- 7479088 TI - A strategy for quickly identifying all unique two-hybrid or library plasmids within a pool of yeast transformants. PMID- 7479086 TI - HMGI(Y) and Sp1 in addition to NF-kappa B regulate transcription of the MGSA/GRO alpha gene. AB - Expression of the chemokine MGSA/GRO is upregulated as melanocytes progress to melanoma cells. We demonstrate that constitutive and cytokine induced MGSA/GRO alpha expression requires multiple DNA regulatory regions between positions -143 to -62. We have previously shown that the NF-kappa B element at -83 to -65 is essential for basal and cytokine induced MGSA/GRO alpha promoter activity in the Hs294T melanoma and normal retinal pigment epithelial (RPE) cells, respectively. Here, we have determined that the Sp1 binding element located approximately 42 base pairs upstream from the NF-kappa B element binds Sp1 and Sp3 constitutively and this element is necessary for basal MGSA/GRO alpha promoter activity. We demonstrate that the high mobility group proteins HMGI(Y) recognize the AT-rich motif nested within the NF-kappa B element in the MGSA/GRO alpha promoter. Loss of either NF-kappa B or HMGI(Y) complex binding by selected point mutations in the NF-kappa B element results in decreased basal and cytokine induced MGSA/GRO alpha promoter activity. Thus, these results indicate that transcriptional regulation of the chemokine MGSA/GRO alpha requires at least three transcription factors: Sp1, NF-kappa B and HMGI(Y). PMID- 7479091 TI - [Computer supported documentation of diagnostic results and interpretation of bone scintigraphy]. AB - A PC software program with a graphical users platform was developed which enables the transfer of bone scintigraphy results to vectorgraphy-based skeleton images, and to implement a fully automatic anatomical localization definition of lesions. Documentation of the results is made possible by a complete, conventional structured medical letter. Additional information such as data on the patient, diagnostic methods and results, sacro-iliacal indexes, recommendations and remarks can be added to the scintigrams. By implementation of a database and PCX interface, the system is open. PMID- 7479090 TI - [Image fusion of MRI and immunoscintigraphy with MAb-170 in ovarian tumors]. AB - In recent years multimodality imaging achieved growing importance. It is mostly performed by means of quite expensive software and hardware solutions. In the present pilot study a simple and low-cost procedure was developed to achieve image fusion in the pelvis. The image data of immunoscintigraphy (SPECT) and MRI were transferred to a personal computer and combined by standard software for image manipulation. The results in eleven patients with space-occupying lesions in the pelvis showed that adequate anatometabolic slices could be achieved. The results show a tendency to increased specificity and precision of multimodality imaging in comparison with SPECT and MRI alone. In conclusion, the low-cost solution, as developed by us, is feasible in clinical practice. Its results are reliable in clinical decision making. PMID- 7479093 TI - [Determination of three body compartments in the chest by scintigraphy]. AB - We adapted a previously described method for the measurement of transthoracic tissue thickness and volumes of blood and interstitial tissue. This non-invasive procedure can be performed safely and by relatively simple means. We used a gamma camera and 99mTc to obtain transmission and emission scintigrams. A computer program written for a personal computer, can recognize the borders of the lung in a fully automatic manner and do the necessary calculations. Clinical studies show significantly higher interstitial volumes of the lung in patients with chronic pulmonary sarcoidosis. PMID- 7479094 TI - [Immunoscintigraphy of breast cancer xenografts. 99m-Tc-labeled anti-mucin monoclonal antibodies BM-7 and 12H12]. AB - The present study was undertaken to evaluate the correlation of the favorable in vitro characteristics of the anti-mucin Mabs 12H12 and BM-7 with high tumor accumulation in vivo. They were labeled with 99mTc; their biodistribution in nude mice bearing mammary tumor xenograft AR was examined and immunoscintigraphy was performed after 24 h. 99mTc-labeling of the Mabs 12H12 and BM-7 led to tumor uptakes of 20.7% and 8.8% ID/g, respectively, after 48 h. Tumor-to-muscle ratios were 31 (12H12) and 18 (BM-7). Tumor xenografts were clearly visualized in immunoscintigrams. Combination of Mab 12H12 and 99mTc provides high tumor-to tissue ratios shortly after administration. PMID- 7479092 TI - [DACS--a cost-advantageous nuclear medical document archiving and communication system]. AB - A simple and inexpensive solution for the complete digital management of a nuclear medical institution is presented. The starting point was provided by five different gamma cameras, one of which was equipped with a PC-based ICON computer. The remaining cameras were first adapted to ICONs and subsequently integrated into a Macintosh LAN, of 15 computers, allowing the entire image acquisition, processing, interpretation, documentation and archiving to be performed on the same comfortable menu-guided surface. Complete patient management is accomplished by a specific commercially available data base to which a specially developed image and document archive was connected. The system described provides complete digital management without the conventional filing systems at the moderate price of approximately DM 11 x 10(4) plus the cost of the new camera computers. PMID- 7479095 TI - [Significance of lymph node metastases in differentiated thyroid carcinoma]. AB - 464 patients with differentiated thyroid carcinoma were scored and treated using a risk-oriented management scheme employing early surgical re-intervention including compartment-oriented lymph-node dissection in cases of lymph-node metastases or local recurrences without routine external radiotherapy. A multivariate analysis showed that there was no effect of lymph-node metastases on survival. The difference in survival between patients with (24%) and without (76%) lymph-node metastases (univariate analysis) were due to the coincidence of higher tumor stage (T4) and higher incidence of peripheral metastases (M1). Thus, provided surgery is efficient, N1 is not a risk factor influencing survival. PMID- 7479096 TI - [Biokinetics of tumor-affinity yttrium preparations--2]. AB - After demonstrating that the application form 87Y-NTA-EDTMP-Ca is superior to the so far used citrate form of yttrium especially regarding the tumour/liver and tumour/bone ratios, the relationships between the concentrations of the components in the 87Y-NTA-EDTMP-Ca mixture and the 87Y biokinetics were investigated in tumour-bearing mice. The increase of the EDTMP and Ca2+ concentrations by several orders of magnitude caused no significant changes of tumour radioactivity but a clear radioactivity decrease in the liver, spleen and bone, respectively. The increase of the NTA concentration led also to a radioactivity decrease in all organs and tissues investigated. A significant radioactivity reduction in the tumour could be observed only after administration of rather high NTA amounts. Considerations of the tumour/background ratios depending on different compositions of the injection mixture showed the possibility of optimizing radionuclide biodistribution. PMID- 7479097 TI - Does every patient deserve a nurse? PMID- 7479098 TI - Nursing in Lithuania as perceived by Lithuanian nurses. PMID- 7479099 TI - Policy-relevant research on infant mortality: rhetorical criticism of mass media. PMID- 7479100 TI - Tuberculosis: what nurses need to know to help control the epidemic. PMID- 7479101 TI - Nurses' attitudes toward active euthanasia. PMID- 7479104 TI - Patient privacy issues in a renal dialysis unit. PMID- 7479103 TI - Professional development. Infection control. Knowledge for practice. PMID- 7479102 TI - New rules for the game: interdisciplinary education for health professionals. PMID- 7479105 TI - Discharge dilemmas. PMID- 7479106 TI - Rationing rationales. PMID- 7479107 TI - Poor performance. PMID- 7479108 TI - Breathing easier. PMID- 7479111 TI - A stereotype too far. PMID- 7479109 TI - Nurses' net assets. PMID- 7479110 TI - Shattered dreams. PMID- 7479112 TI - Setting up an HIV-testing service with same-day results. AB - The advent of human immunodeficiency virus (HIV) has meant that the speciality of genito-urinary medicine has had to respond promptly and decisively in order to limit the extent of infection. This paper describes how a unit has made its service more adaptable to clients' needs to encourage people to come forward for HIV tests. The results of the study reported demonstrate the demand for this service. PMID- 7479113 TI - The resurgence of tuberculosis: current issues for nursing. AB - Completing our series on tuberculosis, this paper explores the extent of and some of the reasons for the recent resurgence of the disease and looks at the role of nurses in responding to the needs of clients. PMID- 7479114 TI - School nurses' response to the measles vaccination campaign. AB - This paper describes the results of a retrospective analysis of the success of nurses, specifically school nurses, in undertaking last year's nationwide rubella and measles immunisation programme for five- to 16-year-olds. The aim of the study was to identify lessons for future practice, training needs, operational planning and resource management. PMID- 7479115 TI - Making sense of drug compliance by patients. AB - Success in the treatment of patients frequently relies on their taking prescribed medication. However, patients do not always comply. Some of the reasons for this are considered together with some of the means by which compliance can be improved. It is suggested that the patient requires sensitive individual attention in order to understand and use his or her medication to its best effect. PMID- 7479116 TI - The role of the neonatal nurse practitioner. AB - The author describes the development of the nurse practitioner role in a neonatal unit. The role of the practitioner and the management of a caseload in collaboration with and under the general supervision of a physician is discussed. This paper continues the more general debate surrounding the development of such roles in other specialties. PMID- 7479117 TI - Community spirit. PMID- 7479118 TI - Bottle-feeding. Formulas and equipment. PMID- 7479119 TI - Telling tales. PMID- 7479120 TI - Risking change in the theatre. PMID- 7479122 TI - Professional development. Infection control. The role of the nurse. PMID- 7479121 TI - From start to finish. PMID- 7479123 TI - Stress in cancer care and coping strategies. PMID- 7479124 TI - A feat of bravery. Interview by David Payne. PMID- 7479125 TI - On duty. Interview by Renata Langford. PMID- 7479126 TI - Effective solutions. PMID- 7479127 TI - The life of Riley. Interview by David Payne. PMID- 7479128 TI - Prone to attack. PMID- 7479129 TI - The use of patient complaints to improve service delivery. AB - In this paper changes to the procedures involved in making complaints are outlined. The alterations in the process follow on from the Department of Health's response to the Wilson report. The new guidelines for dealing with complaints are designed to be simpler and more explicit in order to encourage members of the public to make their concerns known. The ways in which complaints can be used positively to improve on services are outlined. PMID- 7479130 TI - Survival tactics. PMID- 7479131 TI - Complaints can improve care. AB - Complaints are often seen as being entirely negative. However, they could be turned to the advantage of an organisation by allowing it to rectify its shortcomings. A complaints procedure can form part of a quality improvement cycle. This paper discusses why nurses are often unable to see the benefits of complaints and how they can be used to improve patient care in the future. PMID- 7479132 TI - The thyroid gland 2. PMID- 7479133 TI - Political implications of the named-nurse concept. AB - This paper considers the role of the named nurse as a linchpin in the commissioning/providing cycle. It argues that, whatever the impact on patient care, nurses should be wary of claims that the introduction of named nursing through government policy demonstrates esteem for British nursing. PMID- 7479134 TI - Quality of life following bone marrow transplant. AB - Continuing our series on haematological malignancies, this second paper on bone marrow transplantation looks at the psychological impact and quality of life of survivors. It also looks at the effect on patient's sexuality, the psychiatric aspects of BMT and the stress on nursing staff. PMID- 7479135 TI - Badge of courage. PMID- 7479136 TI - Change in consciousness. PMID- 7479137 TI - Home comforts. PMID- 7479139 TI - Core truths. PMID- 7479140 TI - Sensible sex education. PMID- 7479138 TI - Living with the effects of child sexual abuse. PMID- 7479141 TI - A powerful prescription. PMID- 7479143 TI - Moving on up. Interview by Janet Snell. PMID- 7479142 TI - Race relations. PMID- 7479144 TI - Theatre of opportunity. PMID- 7479145 TI - Body of evidence. PMID- 7479146 TI - Breaking the last taboo. PMID- 7479147 TI - Ethnic dilemma. PMID- 7479148 TI - The golden handshake. PMID- 7479149 TI - Recognising and dealing with the abuse of older people. AB - The phenomenon of elder abuse and neglect is not new; indeed, there is evidence that suggests the issues have been present within society for a considerable period of time. Despite the efforts of health- and social-care professionals to draw attention to the topic since the issue was first raised in the late 1970s, it is only since 1988 that the issues related to elder abuse and neglect have begun to be addressed. Health and social-care professionals have not viewed work with older people very positively. PMID- 7479150 TI - A&E--why so complacent? AB - Elder abuse often remains unnoticed in A&E departments. Some reasons lie in the history of care of older people and in the development of elder abuse as a social problem. PMID- 7479151 TI - Sick source of funding. PMID- 7479152 TI - Carers' perspectives. AB - The issues around elder abuse and carers are complex. They concern individual carers and users and their relationships. They concern professionals who are involved and have implications for social policy. Alternatives to family care must be made available and we must address the issue of how long-term care is to be provided. PMID- 7479153 TI - The conflict between lay and professional views of labour. AB - This paper is one of two reviewing the evidence of differing perceptions of pregnancy and labour between medical professionals and the women who are consumers of maternity services, and how those perceptions have changed over the years. The author points out the existence of power imbalances, which can be argued as being at the root of the expert maternity committee's report, Changing Childbirth. The second article will look at the impact of the report on this situation. PMID- 7479155 TI - Meeting mental health needs of ethnic minority groups. AB - The TULIP Outreach Service uses a team approach model to provide community support to people with long-term mental health problems. It was primarily set up to reach people falling through the net of mainstream services, that is, homeless or potentially homeless people and those from minority ethnic groups. This paper describes how the team approach works in practice. It also gives the preliminary results of an evaluation showing the team's success in targeting and supporting these clients. PMID- 7479154 TI - The bladder: cystectomy and ileal conduit to treat cancer. AB - In the fourth paper in this urology series, bladder cancer and its treatment are discussed. The pre- and post-operative care and possible complications are described for the patient undergoing cystectomy and formation of ileal conduit. Psychological care following the stoma formation and the contribution of the stoma therapist are addressed. PMID- 7479156 TI - Beyond belief. PMID- 7479157 TI - Different values. PMID- 7479158 TI - Florence of Arabia. PMID- 7479159 TI - A heart-breaking lesson. PMID- 7479160 TI - Disenchantedly yours. PMID- 7479161 TI - Looking abroad. PMID- 7479162 TI - Enuresis in children. PMID- 7479164 TI - Perspectives on nursing and its role. PMID- 7479163 TI - Professional development. Infection control. Revision notes. PMID- 7479165 TI - Alcohol and alcoholism: role of nursing management. PMID- 7479166 TI - Extroversion, neuroticism and job satisfaction: a comparative study of staff nurses and students. PMID- 7479167 TI - The ECG in Wolff-Parkinson-White syndrome. PMID- 7479168 TI - NASPE Survey on Catheter Ablation. PMID- 7479169 TI - Endocardial fragmented electrogram and prediction of ventricular tachycardia by body surface signal-averaged electrocardiographic mapping. AB - Signal-averaged (SA) electrocardiography and SA electrocardiographic mapping were performed in 50 patients with old myocardial infarction, 19 of whom had left ventricular aneurysm and 11 of whom had clinical sustained ventricular tachycardia. The SA electrocardiogram and SA electrocardiographic mapping data were then compared with those obtained by endocardial catheter mapping in patients with or without fragmented electrograms, sustained ventricular tachycardia, and ventricular aneurysm. Compared to SA electrocardiography, the SA map correlates with sustained VT with improved sensitivity but decreased specificity. However, SA electrocardiographic mapping had the advantage of displaying the extent of the body surface area that was positive for late potentials. In addition, the site of the longest endocardial fragmented electrogram could be predicted by SA electrocardiographic mapping, suggesting that this technique deserves wider clinical application. PMID- 7479170 TI - Initial clinical experience with a new dual sensor SSIR pacemaker controlled by body activity and minute ventilation. AB - Fourteen patients were implanted with a single chamber dual sensor pacemaker (Legend Plus) that measures minute ventilation (VE) via variations in impedance between a bipolar lead and the pacemaker case, and activity via a piezoelectric crystal bonded to the pacemaker case. Chronotropic incompetent patients were exercised on a treadmill and a bicycle in dual sensor mode. Activity only indicated pacing rate was measured using a strap-on pacemaker. Both implanted and strap-on pacemakers were adjusted to yield a steady-state pacing rate of 100 beats/min during hall walk. Pacing rate, VE, and oxygen uptake (VO2) were measured continuously. Linear curve fit analysis slopes for plots of VE versus pacing rate during exercise (1.33-1.49) compared favorably to values reported in normals. Peak pacing rates achieved for treadmill and bicycle testing for dual sensor mode were higher than activity mode alone. Slopes of heart rate to VE or VO2 were not significantly different (P < 0.05) for dual sensor mode in contrast to activity alone. In conclusion, the Legend Plus dual sensor rate adaptive pacing therapy delivered pacing rates more proportional to VE and VO2 under different types of exercise than rates indicated by a strap-on pacemaker in activity mode. PMID- 7479171 TI - Right bundle branch block of unknown age in the setting of acute anterior myocardial infarction: an attempt to define who should be paced prophylactically. AB - It is widely accepted that patients presenting with acute anterior myocardial infarction and acute onset of right bundle branch block should be prophylactically paced in contrast with those who have a chronic bundle branch block. The admitting physician is faced with the dilemma of how to act if the age of this conduction disturbance is unknown. This problem has further intensified in recent years, with the introduction of thrombolytic treatment, where insertion of a central vascular line is associated with increased morbidity. The objectives of this study were to define clinical or electrocardiographic parameters that may help the admitting physician to decide whether patients presenting with an anterior wall myocardial infarction and a right bundle branch block of unknown age should be prophylactically paced. We examined prospectively the in-hospital clinical course of 39 consecutive patients presenting with an acute myocardial infarction in whom the age of a right bundle branch block upon admission was unknown (group C, n = 39) and compared with two similar groups of patients who presented with an acute right bundle branch block (group A, n = 38) and with a known chronic right bundle branch block (group B, n = 22). Thirty-three patients (33%) died, with cardiogenic shock being the leading cause of death in the entire population. Prophylactic pacing, which was carried out in 66% and 54% of patients in groups A and C, respectively, did not reduce mortality rates. No clinical or electrocardiographic variables on admission were predictive to support prophylactic pacing in group C. In 10 of 46 (22%) patients who were prophylactically paced with a transvenous electrode, the following complications attributed to the procedure were detected: (1) either rapid sustained ventricular tachycardia (during implantation) that was unresponsive to overdrive pacing, or ventricular fibrillation necessitating electrical defibrillation (4 patients); (2) recurrent episodes of rapid nonsustained ventricular tachycardia, which stopped only after the pacemaker was turned off (1 patient); (3) complete AV block (1 patient); (4) fever appearing on the third or fourth day after implantation (3 patients); and (4) a large hematoma in the groin in 1 patient who was treated with thrombolysis shortly before pacemaker electrode insertion. Thus, the complications of transvenous temporary pacing in the era of thrombolysis may outweight any theoretical advantage. PMID- 7479172 TI - Pacemaker and defibrillator lead entrapment: case studies. AB - Cadavers and cineradiographic analysis have been used to document the effects of the medial subclavicular musculotendinous complex (MSMC) upon lead function. Four cadavers with pacemakers were dissected and photographed to demonstrate the course a lead takes as it passes through the costoclavicular region. One lead had been placed into the cephalic vein. In the other three cadavers, leads placed by currently accepted techniques of subclavian venipuncture were all found to pass through the soft tissues of the subclavicular region before entering the venous system. Cineradiographic results from a patient with a defibrillator, taken before and after replacement of a broken lead, show the effect of clavicular motion on a lead that passes through the MSMC. Furthermore, cineradiography makes it possible to identify the point where the lead entered the vein, and whether or not it escaped being caught up in the soft tissues of the MSMC. PMID- 7479173 TI - Feasibility of radiofrequency powered, thermal balloon ablation of atrioventricular bypass tracts via the coronary sinus: in vivo canine studies. AB - Radiofrequency catheter ablation of left-sided accessory pathways is technically demanding and usually requires left heart catheterization. The feasibility of creating lesions from within the coronary sinus of sufficient size to ablate accessory pathways in humans using a thermal balloon catheter was studied in 20 dogs. In group 1 (n = 14), 17 thermal inflations were performed in 12 dogs at either 70 degrees, 80 degrees, or 90 degrees C each for 30 or 60 seconds (in 2 dogs two non-thermal control inflations were performed). Animals were sacrificed 6.3 +/- 1.6 days later. In group 2 (n = 6), seven thermal inflations were performed at 90 degrees C each for 180, 300, or 360 seconds. Group 2 animals received antiplatelet and anticoagulant therapy for 1 week and were sacrificed at 13 +/- 10.7 days. In both groups, hemodynamic, angiographic, and electrocardiographic studies were performed at baseline, 1 hour after inflation, and prior to sacrifice. All dogs remained clinically stable throughout the procedure and no complications were attributed to the effect of thermal inflation. Thermal lesions measured 14.4 +/- 4.4 mm in length and extended from the coronary sinus intima to a mean depth of 2.9 +/- 1.2 mm (range 1.4-6.5 mm). Group 2 lesions were significantly deeper than group 1 lesions (P = 0.03). Of the 24 thermal lesions created, atrial necrosis was present in 23 and ventricular necrosis in 11. In all lesions there was some degree of either atrial necrosis, ventricular necrosis, or both. A variable degree of coronary sinus thrombus was present in 18 dogs without clinical sequelae. It is concluded that radiofrequency balloon heating via the coronary sinus can create thermal lesions in the atrioventricular sulcus of dogs that may be of sufficient size to ablate accessory left-sided pathways in humans. PMID- 7479174 TI - Transtelephonic monitoring and transmission of stored arrhythmia detection and therapy data from an implantable cardioverter defibrillator. AB - A new transtelephonic monitoring device designed for use with implantable cardioverter defibrillators (ICDs) was evaluated. It is capable of interrogating ICDs and transmitting the following data via telephone: programmed parameters (e.g., ventricular tachycardia [VT] and ventricular fibrillation [VF] detection, therapies), number of VT and VF episodes, identification of successful therapies, the 20 cycle lengths preceding the last episode detected, the 10 cycle lengths after the last delivered therapy, battery voltage, and real-time transmission of the patient's rhythm. Eighteen patients (mean age 64 +/- 17 years; 15 males) were implanted with an ICD and epicardial lead system. The patients who did not live near the primary hospital were provided with this transmitter and instructed to transmit monthly and whenever presyncope, syncope, or a shock were experienced. Five hundred ten episodes of spontaneous arrhythmia (495 VT, 15 VF) were detected in 14 of 18 patients in a 24-month period and the success of each therapy (antitachycardia pacing, cardioversion 0.4-34 J, defibrillation 34 J) was analyzed. The number of therapies delivered and their success (%) in terminating the arrhythmia were: 380 ramp/86%, 116 burst/84%, 119 cardioversion/57%, and 15 defibrillations/100%. Sixty-three (42%) of the 152 transmissions indicated an arrhythmia. Twenty-five (16%) of the 152 were transmitted because of symptoms. Sixteen (9.7%) of 165 VT episodes could not be terminated by the full set of programmed VT therapies. Analysis of the pre- and post-episode intervals along with the patient's transmitted rhythm indicated that sinus tachycardia or atrial fibrillation were likely responsible for these episodes.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7479175 TI - The subcutaneous array: a new lead adjunct for the transvenous ICD to lower defibrillation thresholds. AB - Despite the benefits of transvenous implantable cardioverter defibrillators (ICDs), concern exists that patients with high defibrillation thresholds (DFTs) have an inadequate safety margin between the DFT and the maximum defibrillator energy. A new transvenous ICD lead adjunct, a subcutaneous lead array (SQ Array), was developed to increase safety margins by lowering DFTs. Composed of three lead elements joined in a common yoke, the SQ Array is tunneled subcutaneously in the left lateral chest. Serving as their own controls, 20 patients were studied intraoperatively comparing transvenous lead-alone DFTs with lead-SQ Array DFTs. Seventeen males and three females were randomized to receive the SQ Array through the CPI Ventak PRx/Endotak 70 series protocol. Mean patient age was 63.7 +/- 2.5 years and mean ejection fraction 0.34 +/- 0.04. DFTs were determine using a precise protocol of step-down/step-up testing commencing at 20 joules. Lead-alone DFTs were tested using the proximal coil as the anode (+). For the lead-SQ Array, the proximal coil and the array were linked as a common anode. The lead-SQ Array resulted in a statistically significant reduction in mean monophasic DFT from 23.3 +/- 2.3 joules (lead-alone) to 13.5 +/- 1.9 joules (lead-SQ Array) (P < 0.001). Six patients had lead-alone DFTs > 25 joules but did not require thoracotomy because of adequate DFT reduction with the SQ Array. We conclude that the SQ Array adjunct to the transvenous ICD lead lowers monophasic DFTs an average of 9.8 joules (40.6%) obviating the need for a thoracotomy in selected patients. PMID- 7479178 TI - Graphical representation of circadian patterns of heart rate variability components. PMID- 7479177 TI - Basic concepts in cellular cardiac electrophysiology: Part I: Ion channels, membrane currents, and the action potential. PMID- 7479176 TI - Effects of nuclear magnetic resonance imaging on cardiac pacemakers. AB - Patients with cardiac pacemakers are currently restricted from nuclear magnetic resonance imaging (MRI). The aim of the study was to analyze the influence of MRI on new generation pacemakers. Tests were performed using a phantom model with seven dual chamber and two single chamber systems in a 0.5 Tesla MRI scanner. Monitoring by telemetry and oscillography were used during the standard clinical scan sequences as well as a pacemaker inquiry after each sequence. Spin echo, gradient echo, and fast field echo sequences were performed with the following stimulation modes: VVI, VVIR, VOO, DDD, DDDR, and DOO. On entering the static magnetic field, the reed switch was activated followed by asynchronous stimulation. The subsequent scan showed no influence on the stimulation function nor on the pacemaker program. Event counter function remained intact. Pacemakers with automatic mode switching to demand pacing or programmed inactivation of the reed switch were triggered in the dual chamber mode and were inhibited in the one chamber mode during the scan. Alterations of pacemaker program or rapid pacing were not observed. MRI scan could induce voltage as high as intracardiac signals, but the stimulation threshold of the heart was not reached. Thus, pacemakers should be programmed in the asynchronous mode during scan to avoid inhibition and trigger mechanism. PMID- 7479179 TI - Specialists and the health care system of the 21st century. PMID- 7479180 TI - To pace or not to pace? PMID- 7479181 TI - Suppression of ventricular output by noise in the atrial channel in a dual chamber pacemaker. AB - In a pacemaker dependent, 60-year-old man with complete heart block, programming high (5.0 V and 1.0 ms) atrial output and 5.0 volt and 0.5 ms ventricular output while in the DDD mode of a pulse generator, resulted in loss of ventricular channel output with resulting asystole, while the simultaneous, pulse generator produced ECG interpretation channel falsely indicated continuing ventricular channel output. Ventricular pacing was promptly restored by programming to the emergency VVI mode. At later operation the atrial lead was determined to be defective, with a low impedance, while the ventricular lead was intact (lead impedance telemetry was not available). The manufacturer has acknowledged a pulse generator design anomaly that may occur in the setting of a low atrial impedance and issued a Health Safety Alert. PMID- 7479182 TI - Cardioverter discharges following sensing of electrical artifact due to fluid penetration in the connector port. AB - We report a unique case of fluid penetration, 3 months after implantation, in the connector port of an automatic implantable cardioverter defibrillator (ICD) with transvenous subcutaneous lead system. The patient had coronary artery disease and recurrent episodes of ventricular fibrillation, the fluid caused electrical signals interpreted as ventricular fibrillation by the device, which triggered shock delivery. PMID- 7479183 TI - Erroneous discharge of an implantable cardioverter defibrillator caused by an electric razor. AB - We report an unusual case of the erroneous discharge of a third-generation multiprogrammable implantable cardioverter defibrillator in a 64-year-old patient with a history of recurrent ventricular tachycardias caused by electromagnetic interference while shaving with an electric razor. Electromagnetic interference was related to a defect in the electrode's insulation and could not be provoked in an intact electrode. PMID- 7479184 TI - Counseling and management of patients with Telectronics 330-801 Accufix leads. PMID- 7479185 TI - X World Symposium on Cardiac Pacing and Electrophysiology. Buenos Aires, Argentina, October 22-26, 1995. Abstracts. PMID- 7479186 TI - Microleakage in Class 2 composite resin restorations. AB - Using the dye penetration method, the effect on the microleakage of class 2 resin restorations of two dentin bonding systems that use 10% citric acid-containing dentin conditions was studied. These results were then compared to the results obtained when the manufacturer's conditioner was replaced by a mild surface active detergent containing 0.2% EDTA and 1% NaF. The cavities that were treated with the 0.2% EDTA-containing conditioner in place of the manufacturer's conditioner showed similar results: Neither of the two bonding systems could totally prevent cervical gap formation. PMID- 7479187 TI - Surface roughness of opalescent porcelains after polishing. AB - Intraoral porcelain polishing is an important consideration in many restorative and esthetic procedures. Several porcelain polishing systems as well as improved ceramics are now commercially available. This study evaluated the efficacy of eight different intraoral polishing techniques on three opalescent porcelains. The surface roughness (Ra) of the opalescent porcelains were measured before and after the polishing procedures with a profilometer. These results were then compared to self-glazed and overglazed control groups. Five of the techniques tested produced surfaces smoother than glazing. The use of a 30-fluted carbide bur before diamond polishing paste produced the smoothest surfaces. PMID- 7479188 TI - Monkey pulpal response to adhesively luted indirect resin composite inlays. AB - Monkey pulpal responses to resin-bonded indirect resin composite inlays were histopathologically evaluated by placing them in either etched-enamel, total etched, or adhesive-resin-lined cavities. Initial pulpal responses caused by re exposure of the cut dentin surfaces and luting procedure subsided if a tight marginal seal was secured by final cementation of the inlay. The adhesive resin coating of freshly cut dentinal walls/floors seems to provide a new technique to protect the dentin and pulp in indirect restorations requiring temporary sealing. PMID- 7479189 TI - In vivo diagnostic assessment of dentinal caries utilizing acid red and povidone iodine dyes. AB - In order to compare the clinical utilization of diagnostic dyes to identify carious dentin, the present in vivo study compared two caries-detector dyes, acid red (1% acid red in propylene glycol wt/wt) and povidone-iodine (8% povidone iodine in water wt/vol). A total of 221 cavities prepared by 19 senior dental students under the supervision of two teachers at the Universidad Nacional Autonoma de Mexico Dental School were used. Dyes were applied double-blind under standardized conditions to at least two teeth of the same patient. Data were analyzed using the Mantel-Haenszel chi-square test and a test of proportions. Results showed that molars or premolars did not appear to be positively stained more frequently by one dye or another. Also, 36.7% of teeth tested positive to either one of the two dyes. Prudent utilization of either acid red or povidone iodine appeared to be equally useful in assisting clinical decisions concerning cavity size while restoring dentinal lesions. However, povidone-iodine may be preferred in clinical settings where acid red is expensive and/or difficult to obtain. Further research is necessary to establish accurately the mechanism of staining by povidone-iodine. PMID- 7479190 TI - The board examination: a true test or only a rite of passage? PMID- 7479191 TI - The use of diet analysis and advice in the management of dental caries in adult patients. PMID- 7479192 TI - Influence of different etchants and etching times on shear bond strength. AB - This study compared the shear bond strength to enamel of Scotchbond Multi-Purpose Dental Adhesive System's bonding resin following etching of enamel with 10% maleic, 1.6% oxalic, 10% phosphoric, and 35% phosphoric acids for 15, 30, and 60 seconds. Three hundred and sixty human molar teeth were used to create 12 groups of 30 enamel specimens per group (n = 30). Flattened enamel surfaces were treated with the different etchants for the time periods indicated, the surfaces primed, and adhesive resin applied according to the manufacturer's recommendations. The specimens were completed with Silux Plus resin and subjected to 1000 thermocycles (5-55 degrees C) followed by shear stress in an Instron Testing Machine to failure within a 24-hour period. A two-way ANOVA revealed significant differences for shear bond strength values by type of etchant (10% phosphoric, 35% phosphoric > 10% maleic > 1.6% oxalic acid) (P < 0.005) and by length of application time (P < 0.005). The interaction term for these two treatments was statistically significant (P < 0.005). PMID- 7479193 TI - Revised FDA labeling guideline for theophylline oral dosage forms. PMID- 7479195 TI - Bioequivalence and narrow therapeutic index drugs. AB - Every prescription written for a generic drug requires an act of faith by the prescriber that any one of the several available products will be therapeutically equivalent to the innovator (brand name) products. Concerns about this act of faith have been expressed for many years, particularly in the wake of the generic scandals that occurred in 1989-1990, and especially relative to the drugs with a narrow therapeutic range. We contend that these drugs are actually the least likely to pose problems in ensuring therapeutic equivalence, but that new criteria must be established for bioequivalence because the present system is wasteful and is stifling innovation in the industry. We propose four suggestions to the scientific and regulatory communities that we believe could assist in modifying the process such that innovation is encouraged and practitioners are reassured relative to the appropriateness of using generic drugs. PMID- 7479194 TI - A large population-based follow-up study of trimethoprim-sulfamethoxazole, trimethoprim, and cephalexin for uncommon serious drug toxicity. AB - We conducted a population-based 45-day follow-up study of 232,390 people who were prescribed trimethoprim-sulfamethoxazole (TMP-SMZ), 266,951 prescribed trimethoprim alone, and 196,397 prescribed cephalexin, to estimate the risk of serious liver, blood, skin, and renal disorders resulting in referral or hospitalization associated with these drugs. The results were based on information recorded on office computers by selected general practitioners in the United Kingdom, together with a review of clinical records. The risk of clinically important idiopathic liver disease was similar for persons prescribed TMP-SMZ (5.2/100,000) and those prescribed trimethoprim alone (3.8/100,000). The risk for those prescribed cephalexin was somewhat lower (2.0/100,000). Only five patients experienced blood disorders, one of whom was exposed to TMP-SMZ; of seven with erythema multiforme and Stevens-Johnson syndrome, four were exposed to TMP-SMZ. The one case of toxic epidermal necrolysis occurred in a patient who took cephalexin. Finally, only five cases of acute parenchymal renal disease occurred, none likely to be caused by a study drug. We conclude that the risk of the serious diseases studied is small for the three agents, and compares reasonably with the risk for many other antibiotics. PMID- 7479196 TI - Thrombolytic-associated cholesterol emboli syndrome: case report and literature review. AB - Thrombolytics can cause cholesterol embolization syndrome (CES). This adverse effect has received less attention than other risks of thrombolytic therapy, such as systemic bleeding and hemorrhage, with only sporadic reports of CES in the literature. Risk factors have not been consistently identified and emphasized; therefore, occurrence of CES after thrombolysis remains difficult to predict, it results in substantial morbidity and mortality, and it lacks effective pharmacologic treatment. Heightened awareness of the disorder can aid in its correct identification and reporting. PMID- 7479197 TI - Pharmacokinetics and pharmacodynamics of high-dose zidovudine administered as a continuous infusion in patients with cancer. AB - STUDY OBJECTIVE: To investigate the pharmacokinetics and pharmacodynamics of high dose intravenous zidovudine (ZDV). DESIGN: Phase 1, dose-escalating, unblinded study in patients with cancer. SETTING: A university-affiliated cancer treatment center. PATIENTS: Fourteen patients (6 women) with solid tumors that were unresponsive to standard therapy received 31 courses of therapy. INTERVENTIONS: Intravenous ZDV was administered in doses of 2, 3, 4, 5.5, 7, 8.5, 10, 12, 15, or 20 g/m2/day as a continuous infusion over 48 hours. Patients also received fluorouracil plus leucovorin for 24 hours before the start of and during the ZDV infusion. If no dose-limiting toxicities were encountered, subsequent doses were escalated. Blood samples were collected at 24 and 48 hours after the start of the infusion, and hourly for 4 hours after stopping the infusion. Urine was collected in five patients during the infusion and for 24 hours after stopping it. Blood for measuring peripheral white blood cells was collected before and at the end of the infusion in seven patients to measure DNA chain breaks due to incorporation of ZDV. MEASUREMENTS AND MAIN RESULTS: Zidovudine was measured in plasma by high performance liquid chromatography and in urine fluorescence polarization immunoassay. Its incorporation into DNA was measured by determining DNA strand breakage in peripheral white blood cells using fluorescence analysis. Pharmacokinetic models were fit to plasma ZDV concentrations using extended least squares regression. Short-term high-dose ZDV was generally well tolerated, with adverse effects related to large amounts of free water administered during the infusion. The mean (SD) ZDV pharmacokinetic values were total clearance 1.44 (1.09) L/hr/kg, volume of distribution 2.72 (2.97) L/kg, and half-life 1.2 (0.6) hours. There was considerable interpatient variability in total drug clearance. Although ZDV exposure increased proportionately with increasing dose, two of three patients receiving the highest dose (20 g/m2/day) had markedly low total drug clearances. The relation between the percentage of abnormal DNA in peripheral white blood cells and zidovudine area under the plasma ZDV versus time curve was described by the Emax pharmacodynamic model. CONCLUSIONS: The pharmacokinetics of high-dose ZDV administered by continuous infusion to patients with cancer are similar to those reported with lower doses in patients with infection due to the human immunodeficiency virus. Further study of potential nonlinear pharmacokinetic behavior at doses above 20 g/m2/day is necessary. The high between-patient variability in ZDV clearance results in variable levels of exposure in vivo, and indicates the need for concentration- or effect-controlled study designs in the further evaluation of the agent's antineoplastic effects. PMID- 7479198 TI - The effect of high-dose ascorbate supplementation on plasma lipoprotein(a) levels in patients with premature coronary heart disease. AB - STUDY OBJECTIVE: To determine the efficacy of high-dose ascorbate supplementation in lowering lipoprotein(a) [Lp(a)] levels in patients with premature coronary heart disease (CHD). DESIGN: Randomized, double-blind, placebo-controlled trial. SETTING: Outpatient clinic. PATIENTS: Forty-four patients with documented premature CHD, defined as confirmed myocardial infarction and/or angiographically determined stenosis of 50% or greater in at least one major coronary artery before age 60 years. INTERVENTIONS: Patients were block randomized on the basis of age, gender, and screening Lp(a) concentrations to receive ascorbate 4.5 g/day or placebo for 12 weeks. MEASUREMENTS AND MAIN RESULTS: High-dose ascorbate was well tolerated and produced a marked elevation in mean plasma ascorbate levels (+1.2 mg/dl; p < 0.001). Multiple linear regression analysis revealed no significant effect of supplementation on postintervention Lp(a) levels (p = 0.39) in a model that included treatment group assignment, and baseline Lp(a) levels. CONCLUSIONS: Our findings do not support a clinically important lowering effect of high-dose ascorbate on plasma Lp(a) in patients with premature CHD. PMID- 7479199 TI - Pharmacokinetics and pharmacodynamics of glipizide after once-daily and divided doses. AB - STUDY OBJECTIVE: To determine the pharmacokinetics and pharmacodynamics of glipizide given as a single, oral, 20-mg dose, versus three different divided dose regimens totaling 20 mg each. DESIGN: Randomized (in order of dosing regimens), open-label, crossover study. SETTING: University medical center clinical research center. PATIENTS: Six subjects with noninsulin-dependent diabetes mellitus. INTERVENTIONS: Patients were studied on four separate occasions separated by at least 3 days. The divided-dose regimens were designed to simulate delayed absorption of the drug over 2, 4, and 8 hours. Blood samples for measuring glipizide, glucose, and C-peptide were obtained over 24 hours. MEASUREMENTS AND MAIN RESULTS: Glipizide peak concentrations and time to peak differed significantly with the dosage schedule; when smaller doses were administered more often, peak concentrations were lower and more delayed. The mean values for area under the curve from time zero to infinity (range 7240.7 10,001.8 micrograms.L-1.hr-1; 16,226-22,414 nmol.L-1.hr-1), clearance (0.44-0.64 ml.min-1.kg-1; 0.07-0.11 ml.sec-1.kg-1), post-distribution phase volume (0.17 0.25 L.kg-1), and half-life (4.2-5.4 hrs) were not significantly different among regimens. Neither morning fasting glucose nor maximum and minimum times and concentrations of glucose and C-peptide over 24 hours were statistically different among regimens. Similarly, no significant differences were found in area under the concentration-time curve for glucose and C-peptide measured over 2.5 hours after each meal and from time zero to 24 hours. CONCLUSIONS: The timing of a glipizide dose in relation to a meal and simulated delayed or prolonged absorption appear to have little influence on the drug's pharmacodynamic effects. PMID- 7479200 TI - Piperacillin-tazobactam pharmacokinetics in patients with intraabdominal infections. AB - STUDY OBJECTIVE: To determine the appropriate compartmental and noncompartmental pharmacokinetic parameters for intravenous piperacillin and tazobactam. DESIGN: Sequential selection of patients entered into a randomized, open-label clinical efficacy trial. SETTING: Los Angeles County-University of Southern California Medical Center. PARTICIPANTS: Sequential sample of 18 patients admitted for intraabdominal infections and consented into a comparative antibiotic trial. INTERVENTIONS: Patients received piperacillin 4 g plus tazobactam 500 mg by intravenous intermittent infusion every 8 hours. MEASUREMENTS AND MAIN RESULTS: The estimated noncompartmental pharmacokinetic parameters (mean +/- SD) for piperacillin and tazobactam, respectively, were as follows: maximum concentration in plasma 218.7 +/- 48.9 micrograms/ml and 27.8 +/- 9.1 micrograms/ml; half-life 1.07 +/- 0.22 hours and 1.00 +/- 0.27 hours; elimination rate constant 0.67 +/- 0.13 hr-1 and 0.73 +/- 0.18 hr-1; area under the concentration-time curve from zero hour to infinity 288.5 +/- 71.25 mg.hr/L and 36.3 +/- 9.55 mg.hr/L; total plasma clearance 14.75 +/- 3.93 L/hour and 14.78 +/- 4.39 L/hour; renal clearance 5.69 +/- 1.94 L/hour and 7.85 +/- 3.37 L/hour; volume of distribution at steady state 21.00 +/- 4.18 L and 22.47 +/- 8.27 L; and mean residence time 1.72 +/- 0.29 hours and 1.79 +/- 0.35 hours. CONCLUSION: Our findings were similar to those in other surgical patient models. The two-compartmental model best described piperacillin and tazobactam disposition in our patients. Bayesian analyses of the two-compartment models of piperacillin and tazobactam were able to predict trough, peak, and 2-hour postadministration levels without bias. PMID- 7479202 TI - Publication rates in U.S. schools and colleges of pharmacy, 1976-1992. AB - To establish publication rates of U.S. schools and colleges of pharmacy (SCOP) for 1976-1992, we obtained data from the Science Citation Index (SCI) Corporate Index. The SCI data base covers the top 4500 journals in the technical and scientific fields. Citations were counted without regard to publication type (letter, abstract, review, etc.). Duplicative publications were eliminated. Faculty counts were obtained from the American Association of Colleges of Pharmacy Roster of Faculty and Staff for the inclusive years. Total publications for all schools increased over 100% from 1976 to 1992, and the number of faculty members increased by 40% during that time. However, only 12 (16%) of the SCOP averaged 50 or more publications/year, whereas 43 (59%) averaged fewer than 20. Data were also normalized by full-time faculty members. Only 13 (18%) SCOP averaged 1.0 or more publication/faculty/year, and 38 (52%) averaged fewer than 0.5. Publication rates were greater for medical center-based than for nonmedical center-based SCOP (p < 0.05), and for public than for private SCOP (p < 0.05). These data suggest that over half of the existing SCOP are minimally productive, generating less than 20 publications/year or 0.5 publication/faculty/year. PMID- 7479201 TI - Comparison of the pharmacodynamic activity of cefotaxime plus metronidazole with cefoxitin and ampicillin plus sulbactam. AB - STUDY OBJECTIVE: To compare the pharmacokinetic and pharmacodynamic activity of three drug regimens: cefotaxime plus metronidazole, cefoxitin, and ampicillin sulbactam against two organisms frequently isolated in intraabdominal infection, Escherichia coli and Bacteroides fragilis. DESIGN: Open-label, three-way crossover study. SETTING: Hartford Hospital Clinical Research Center. PARTICIPANTS: Nine healthy volunteers. INTERVENTIONS: Subjects received the following regimens: (1) a single 1-g intravenous dose of cefotaxime plus a single 500-mg oral dose of metronidazole; (2) two intravenous doses of cefoxitin, 2 g each dose given every 6 hours; and (3) two intravenous doses of ampicillin sulbactam, 3 g each dose given every 6 hours. MEASUREMENTS AND MAIN RESULTS: Serum bactericidal titers and drug concentrations were measured over a 12-hour period. The cefotaxime-metronidazole regimen showed superior activity against E. coli compared with ampicillin-sulbactam and cefoxitin. The mean areas under the bactericidal activity curve (AUBC) for the three regimens were 550.2, 68.7, and 48.9, respectively (p = 0.0001). There was no significant difference in AUBC among the three regimens for B. fragilis. Serum concentrations of cefotaxime remained above the minimum inhibitory concentration (MIC) for E. coli significantly longer than did concentrations of ampicillin-sulbactam and cefoxitin (p = 0.0002 and p = 0.0023, respectively). Serum concentrations of metronidazole were still at 9 times the MIC for B. fragilis at the end of the 12 hour dosing interval; for ampicillin-sulbactam and cefoxitin concentrations remained above the MIC for one-half and less than one-fourth, respectively, of the dosing interval (p < 0.0001). The ratio of AUC:MIC was also favorable for metronidazole (212.2) compared with 63.4 for ampicillin-sulbactam and 9.2 for cefoxitin. CONCLUSIONS: The combination of cefotaxime-metronidazole, even at the relatively low doses used in this study, provides coverage against gram-negative and anaerobic pathogens that is at least as effective as that of cefoxitin and ampicillin-sulbactam. In addition, its cost is considerably less expensive than that of the other regimens. PMID- 7479203 TI - Psychotropic drug use in relation to psychiatric symptoms in community-living persons with Alzheimer's disease. AB - We attempted to determine the relationship between psychiatric symptoms and psychotropic drug use in persons with Alzheimer's disease based on a multicenter patient registry of 671 community-living persons diagnosed with the disease by published criteria. Logistic regression was performed to determine which symptoms were associated psychotropic use after controlling for age, sex, and Mini-Mental Status Examination (MMSE) score. At least one psychotropic drug was reported by 31% of patients, and 66% had at least one psychiatric symptom. Antipsychotics were associated with a lower MMSE score (odds ratio = 0.92, 95% confidence interval 0.88-0.97), emotional lability (OR = 4.52, 95% CI 1.69-11.94), and hallucinations (OR = 6.54, 95% CI 2.99-14.26). Antidepressants were associated with depressive symptoms (OR = 5.8, 95% CI 2.61-13.46), and benzodiazepines with a lower MMSE score (OR = 0.93, 95% CI 0.90-0.97). Community-living persons with Alzheimer's disease are frequently prescribed psychotropic drugs; however, more than 50% of patients with a psychiatric symptom did not report taking one of these agents. This suggests that alternative therapies and no treatment are also prevalent. PMID- 7479204 TI - The effect of age and everyday exercise on steady-state plasma digoxin concentrations. AB - Age-related physiologic changes may significantly alter the disposition and pharmacologic characteristics of many drugs. The elderly are the most frequent users of digoxin because of increased prevalence of atrial fibrillation and congestive heart failure. This study was conducted to confirm the decrease in digoxin concentrations during exercise, to determine if age is a factor in this decrease, and to explore the difference between chronologic age and physiologic age. Eighteen men age 50-85 years were treated with digoxin for more than 1 month before enrolling and had serum digoxin concentrations of 0.4-2.0 micrograms/L. They were evaluated during a 3-hour period in the morning (A.M. dose withheld). Blood samples were obtained every 10 minutes during sequential 60-minute periods of rest (phase I), walking (phase II), and rest (phase III). There were no significant differences in mean concentration between phases II and I (p < 0.76), phases III and phase I (p < 0.70), or phases II and III (p < 0.37). The effect of age was positively correlated with the mean concentration of phase II but was not statistically significant (p < 0.62). Statistically significant correlations were seen only between the exercise phase and serum albumin and Mini-Mental Status Examination scores. We conclude that exercise has minimal, if any, clinically relevant effects on plasma digoxin concentrations. Increasing chronologic age has no influence on a decrease in the concentrations with exercise; a younger physiologic age may play a role. PMID- 7479205 TI - Pharmacodynamic modeling of finasteride, a 5 alpha-reductase inhibitor. AB - Finasteride is a 4-azasteroid inhibitor of one isoenzyme of 5 alpha-reductases that converts testosterone to dihydrotestosterone (DHT). We characterized the time course of DHT concentrations. The following model was used to assess DHT pharmacodynamics: [formula: see text] where joint fitting of three dose levels yielded kin0 = 28% change/hour, kout = 0.28 hour-1, IC50 = 0.012 ng/ml, and Emax = 0.7. The modification of a previous model with the maximum partial effect factor, Emax, may be useful in characterizing the pharmacodynamics of drugs with similar indirect mechanisms. PMID- 7479206 TI - Computer-assisted continuous infusion for the delivery of target-controlled infusions of propofol during outpatient surgery. AB - Propofol was administered intravenously using a first-order, three-compartment, computer-controlled infusion pump (CCIP) based on the Ohmeda 9000 syringe pump. The CCIP system produced a target-controlled infusion (TCI) based on the estimated blood concentration (EBC). Twenty-five patients undergoing ambulatory surgery completed the study. The induction EBC was 4.72 +/- 0.28 micrograms/ml, and was achieved rapidly with minimal hemodynamic changes. The emergence EBC was 1.55 +/- 0.21 micrograms/ml, and the discharge EBC was 1.06 +/- 0.22 micrograms/ml. The EBC, together with clinical signs, allowed for predictable inductance and emergence from anesthesia using propofol. PMID- 7479207 TI - Heparin monitoring in acute thrombosis associated with antiphospholipid antibody syndrome. AB - Antiphospholipid antibody syndrome is associated with recurrent thromboembolism and the presence of antibodies directed against negatively charged phospholipids. Despite significant hypercoagulability, patients with the disorder often have elevated baseline activated partial thromboplastin times. Although heparin is indicated for the treatment of acute thrombosis, heparin monitoring is complicated by laboratory test interference. A practical treatment approach involves patient-specific screening for a reagent insensitive to the presence of the inhibitor, and determination of a reagent-specific therapeutic range in seconds that corresponds to heparin serum concentrations of 0.2-0.4 U/ml. Other monitoring methods, including using the patient's baseline activated partial thromboplastin time or using an antifactor Xa activity assay, have not been reported or studied but may offer alternatives for heparin monitoring in patients with coagulation laboratory test interference associated with antiphospholipid antibody syndrome. PMID- 7479208 TI - Acetazolamide toxicity and pharmacokinetics in patients receiving hemodialysis. AB - Acetazolamide-induced central nervous system toxicity occurred in two patients undergoing hemodialysis. Symptoms of toxicity included fatigue, lethargy, and confusion, which resolved several days after discontinuing acetazolamide. Pharmacokinetic studies showed markedly elevated serum concentrations of the drug during the period of toxicity, which decreased at a slower rate compared with that reported in patients with normal renal function. The effect of hemodialysis on acetazolamide clearance was quantified. The agent should be avoided in patients receiving dialysis unless the dosage is reduced and serum concentration monitoring can be performed in a timely manner. These patients should be monitored closely for central nervous system toxicity if acetazolamide is given. PMID- 7479210 TI - Atracurium resistance in a critically Ill patient. AB - A previously healthy 25-year-old man with metastatic testicular teratocarcinoma became resistant to atracurium-induced neuromuscular blockade as evidenced by train-of-four (TOF) monitoring combined with clinical assessment. Subsequently he had an adequate response with a standard dosage of pancuronium. During the first 10 days of neuromuscular blockade, the atracurium requirements escalated from 0.31 to 1.7 mg/kg/hour, guided by TOF monitoring, movement, and spontaneous respirations. The infusion was discontinued but later reinstituted. Despite a total atracurium loading dose of 1.4 mg/kg followed by an infusion rate titrated to 1.7 mg/kg/hour, inadequate paralysis persisted. Atracurium was terminated and an intravenous infusion of pancuronium 0.10 mg/kg/hour was started. Over the next 3 days the pancuronium infusion was titrated down to a range of 0.04-0.06 mg/kg/hour, followed by a maintenance infusion of 0.01-0.05 mg/kg/hour for 5 days. A pharmacokinetic alteration, such as increased metabolism or elimination, may have caused the atracurium resistance. PMID- 7479209 TI - Amoxapine overdose in a young man: a transient mitochondrial abnormality? AB - Amoxapine is a second-generation tricyclic antidepressant structurally related to the neuroleptic loxapine. It was previously marketed as an alternative to traditional tricyclic antidepressants because of alleged shorter onset of action and fewer cardiotoxic effects. However, various adverse reactions, including cardiac dysrhythmias, renal failure, coma, seizures, and neuroleptic malignant syndrome, were reported during therapy or after acute overdose. A 14-year-old boy ingested 1900 mg of amoxapine and developed seizures, hypertension, hyperpyrexia, altered mental status, myoglobinuria, renal failure, and transient magnetic resonance imaging (MRI) changes suggestive of hypertensive encephalopathy and neuroleptic malignant syndrome. Since mitochondrial disorders can cause multisystem failure, including encephalopathy, renal tubular dysfunction, and myopathy, a transient, toxic disorder of mitochondrial function was considered as the basis for the patient's clinical and MRI changes. PMID- 7479211 TI - Inconsistency with train-of-four monitoring in a critically ill paralyzed patient. AB - Problems occurred with train-of-four (TOF) monitoring during prolonged therapy with nondepolarizing neuromuscular blocking agents (NNMBAs). A previously healthy 25-year-old male with metastatic testicular teratocarcinoma was paralyzed with an atracurium infusion to facilitate mechanical ventilation. Dosage titration was initially based on clinical assessment; however, on day 4 of atracurium, TOF monitoring was initiated. During days 4 to 10 of atracurium therapy, TOF monitoring correlated well with clinical assessment of the depth of paralysis. On day 13, atracurium was discontinued and a pancuronium infusion was initiated. During the 9 days of pancuronium therapy, TOF monitoring suggested overparalysis on several occasions (no thumb twitch at 80 mamp of ulnar nerve stimulation) despite clinical evidence of spontaneous movement or respirations. The patient was edematous and had extremely dry skin during some of these instances of inappropriate TOF response. Although these problems were rectified, TOF response continued to be erroneous. Thus we had to rely primarily on clinical assessment to monitor the duration of NNMBA therapy. This case demonstrates that TOF data and clinical assessment of neuromuscular blockade may not always correlate. PMID- 7479212 TI - Train-of-four: to use or not to use. AB - A pattern of nerve stimulation called train-of-four (TOF) is frequently used to monitor therapy with nondepolarizing neuromuscular blocking agents in patients treated in intensive care units. Based on our experience with TOF monitoring in several critically ill patients, we believe its application as an indicator of neuromuscular blockade may be problematic in this setting. PMID- 7479213 TI - [Selected aspects concerning the possibility of improving treatment results for malignant neoplasms]. PMID- 7479214 TI - [Evaluation of thyroid anti-membrane and anti-thyroglobulin antibody determination in patients with thyroid cysts treated with external drainage under ultrasonographic control]. AB - In patients (43 women and 12 men) external open drainage under ultrasonography control was performed. In 25 cases contents of serous nature, in 27 cases of colloid and in 3 cases of suppurative character were obtained. Anti-membrane thyroid antibodies (ATMA) and anti-thyroglobulin antibodies (Tg-Ab tire) were determined in the cyst contents as well as in the serum. The antibodies were present in the serum of 48% of the patients with serous cyst and in 52% with colloidal cysts. In serous cyst contents, ATMA and Tg-Ab antibodies were present in 52% of cases, while in the colloidal contents ATMA antibodies were found in 60% of patients and Tg-Ab antibodies in 68%. In 3 patients with cysts containing pus no antibodies were found. When antibodies were detected in the contents of serous and colloidal cysts their presence were also found in the serum. Among 55 cases with thyroid cysts, the antibodies were found in the serum of 47% of patients. The results indicate involvement of immunological factors the role of which has not been explained. PMID- 7479215 TI - [Concentration of chromium in blood serum of patients with chronic renal failure]. AB - A group of 66 patients with chronic renal failure (CRF) was studied. 42 of them (21 to 63 years old) were treated with haemodialysis and 24 of them (20 to 63 years old) were treated without dialysis. A group of 16 healthy subjects, 19 to 73 years old, were also studied. The blood samples in non-hemodialyzed patients and healthy subjects were taken only once after cannula had been inserted into the antebrachial vein. The blood samples in hemodialyzed patients were withdrawn four times: just before dialysis, during haemodialysis after the first hour of dialysis (from the arterial and from venous line of the dialyser) and after haemodialysis. Chromium concentrations in serum in all examined groups and chromium concentrations in the dialysis fluid as well as in the demineralized water used for dialysis were measured by flame atomic absorption spectrophotometry. In all patients with CRF serum chromium concentration was significantly higher than in the controls. During haemodialysis a decrease of serum chromium concentration was observed. Significant correlations between chromium concentration and creatinine concentration, haemoglobin concentration, haematocrit value, erythrocytes were observed. CONCLUSIONS: 1. Serum chromium concentration in patients with CRF was significantly higher than in normals. 2. No significant differences were found in serum chromium concentration between subgroups of patients with chronic renal failure. 3. Haemodialysis decreases significantly serum chromium concentration. PMID- 7479216 TI - [The influence of recombinant human erythropoietin (rHu-EPO) on circadian rhythm of prolactin secretion in patients with chronic renal failure treated with hemodialysis]. AB - Patients with chronic renal failure (CRF) have high serum prolactin level. In healthy subjects the circadian rhythm of prolactin secretion is well known. This study aimed to investigate, if the circadian rhythm of prolactin was maintained in patients with CRF treated with haemodialysis despite hyperprolactinaemia, and how the treatment with erythropoietin influenced the circadian rhythm of prolactin. Serum prolactin was determined four times a day at 6:00, 12:00, 18:00, 24:00 in 30 patients with CRF and in 15 healthy subjects. In the patients with CRF this investigation was both done on the day with haemodialysis and on the day without haemodialysis. The patients were divided into two groups: the first one included patients treated with erythropoietin and the second one included patients not treated with erythropoietin. The results of the investigation were analysed using the Cosinor test. Serum concentration of hormones was determined by LIA. It was shown that the circadian rhythm of prolactin was maintained in patients with CRF both on the day with and without haemodialysis. Both in patients with CRF and in healthy subjects the circadian rhythm indicated the highest serum concentration of prolactin in the night hours i.e. 22:30-1:00. In patients treated with erythropoietin a significantly lower average day concentration of prolactin was observed in comparison with patients not treated with erythropoietin. PMID- 7479217 TI - [The effect of short-term erythropoietin therapy on selected parameters of cell mediated and humoral immunity in hemodialyzed uremic patients]. AB - This study aimed to assess the influence of short-term therapy with recombinant human erythropoietin (rhEPO) on selected parameters of humoral and cell mediated immunity in haemodialyzed uraemic patients (HDP). In 12 HDP before, and after 1 and 3 months of rhEPO therapy the following parameters were assessed: nitroblue tetrazolium (NBT) test, NBT test after latex stimulation, number of B, T and CD4 and CD4- and CD8-positive T lymphocytes, serum concentrations of IgG, IgA and IgM. The number of granulocytes with a positive NBT test was significantly higher after 3 months of rhEPO therapy. The number of granulocytes with a positive NBT test after latex stimulation increase both after 1 and 3 months of rhEPO therapy but significantly only after 3 months of treatment. The number of CD4-positive T lymphocytes increased significantly after 3 months of rhEPO therapy, while the number of CD8-positive lymphocytes decreased significantly after 1 month of therapy. The CD4/CD8 ratio increased significantly after 3 months of rhEPO therapy. Serum IgA concentration increased significantly after 1 and 3 months, while serum IgG level only after 3 months of rhEPO therapy. From the results obtained in this study it follows that rhEPO therapy exerts a positive effect on function of both T and B lymphocytes in haemodialyzed uraemic patients. PMID- 7479218 TI - [Sick sinus syndrome (extrinsic and intrinsic form) during long term observation]. AB - The aim of the study has to be find, on basis of long term follow-up, an independent factors determining of pacemaker implantation in patients with intrinsic and extrinsic form of sick sinus syndrome. The second point was to find an independent risk factors of stroke and another serious complication in this group of patients (specially in paced group). The study group consisted 169 pts with sick sinus syndrome (in all pts abnormal electrophysiological tests- transoesophageal atrial stimulation). The form of sinus node dysfunction (extrinsic or intrinsic) was determined on basis of electrophysiological study after pharmacological denervation of the heart (propranolol and atropine i.v. in doses 0.2 and 0.04 mg/kg body weight respectively). The statistical analysis was performed on all pts (97 men and 72 women 48 +/- 16 yrs old) using Cox's model of proportional hazardous analysing. The follow-up period ranging from 14 to 84 months (mean 49.9 +/- 19). Pacemaker was implanted in 88 pts during observation period. Stroke signs were found in 7 pts including 6 pts after pacemaker implantation. There was 13 death (7 sudden or primary neurological death). CONCLUSIONS: 1) independent factors of pacemaker implantation were syncope, age and persistent bradycardia, 2) form of sinus node dysfunction was not an independent factor of this decision, 3) age (older), persistent after pacemaker implantation syncope and registered in transesophageal ecg ventriculo-atrial condition were independent risk factors of stroke in pts with VVI stimulation, 4) despite of pacemaker implantation (VVI mode) many patients have presyncopal or syncope attacks but life comfort in this group was much better then before implantation. PMID- 7479219 TI - [Comparison of selected parameters of dialysis quality in hemodialysis patients not requiring and requiring treatment with erythropoietin]. AB - After 1/2 year long clinical observation 50 maintenance hemodialyzed patients were divided into two groups. Group I consisted of patients with haemoglobin concentration exceeded 9.5 g/dl and hematocrit above 30%, not treated with rHuEPO. Patients of group II permanently or periodically required treatment of rHuEPO to maintain haematological parameters written above. Then mean maintaining dose of rHuEPO was 5,500 units weekly. The concentrations of haemoglobin, albumin, creatinine, transferrin, KT/V and body mass index (BMI) handled as parameters of quality of dialysis therapy (PQD) of two groups were compared. Lack of treatment with rHuEPO in hemodialyzed patients of group I, with adequate Hb concentration, does not change significantly PQD in comparison with patients of group II treated with rHuEPO. PMID- 7479220 TI - [Pulmonary function in patients with rheumatic mitral valve disease. Preoperative evaluation]. AB - The aim of the study was to observe the mechanism of cardiac-airway interaction in patients with rheumatic mitral valve disease. 61 pts with combined mitral disease and 9 with isolated mitral stenosis (42 females, 28 males; mean age 47 y) were studied. They were in NYHA functional class II-IV (36 pts in II, 22 in III, 12 in IV class). Each patient was assessed by clinical, noninvasive cardiological investigation (estimation of left atrium dimension, mitral gradient, mitral area and right ventricular systolic pressure on echocardiographic examination and relative heart volume on fluoroscopy) and lung function studies i.e. spirography, plethysmography, flow-volume curve and histamine provocation test. The bronchial hyperreactivity was found in 20 pts (75% undergoing test-mean PC40 = 0.1474 mg/ml). In 7 pts histamine test was negative. The reversible airway obstruction (after 200 micrograms FNT) was observed in 43 (61%) pts. CONCLUSION: In pts with mitral valve disease we observed significant bronchial hyperresponsiveness to histamine and reversible airway obstruction with no correlation between pulmonary function parameters and cardiological status. PMID- 7479221 TI - [Unusual clinical course of ruptured aortic aneurysms--report of three cases]. AB - Three cases of unexpected clinical course of ruptured aortic aneurysm have been presented in patients of their 7th decade life. All of them had arterial hypertension. Signs and symptoms on admission to hospital (dysphagia, chest and interscapular pain, hematemesis, abdominal pain, elevated body temperature and diminished exercise tolerance) were non-specific of aortic aneurysm, suggesting other disease. Dramatic clinical course with hypovolemic shock in two cases led to death. One of them refused surgery. In the third one, in spite of blood effusions to pleural cavity, pericardial sac and mediastinum, effective hypotensive therapy with a preservation of the slow heart rate and fluid evacuation from pericardial sac, gave the opportunity to perform elective surgery. Aortic dissection often presents an atypical course and when suspected, all available imaging technics including computed tomography and nuclear magnetic resonance must be used. PMID- 7479222 TI - [Treatment of multiple myeloma associated anemia with human recombinant erythropoietin in a hemodialysis patient]. AB - Recombinant human erythropoietin (rHuEPO) was used to treat the anaemia in a hemodialyzed 60 year old man with multiple myeloma requiring chemotherapy; the type of serum M component was IgA kappa. During the 5 months before rHuEPO treatment the patient was given multiple blood transfusions. rHuEPO was started 125 U/kg/week subcutaneously in divided doses after dialysis. After introduction of rHuEPO, blood transfusions were no longer required and haemoglobin concentration increased from 5.2 mmol/l to 6.9 mmol/l after 8 weeks of treatment. We conclude that rHuEPO treatment in haemodialysed patients with multiple myeloma may be safe, well tolerated and clinically effective. PMID- 7479223 TI - [Vasculitis--new etiopathogenic and clinical aspects]. PMID- 7479224 TI - [XXI International Congress of Electrocardiology 3-7 July 1994, Yokkohama, Japan]. PMID- 7479225 TI - [Effect of pharmacologic blockade of the autonomic system on time domain parameters of atrial and ventricular signal-averaged ECG in subjects with mitral valve prolapse]. AB - The aim of this study was evaluation of pharmacologic autonomic blockade on time domain parameters of atrial (ASAECG) and ventricular signal-averaged ECG (SAECG) in patients with mitral valve prolapse syndrome (MVP) regarded 20 subjects with MVP (mean age 34.9 +/- 5.3) (group I) were studied as well as a control group consisting of 20 healthy subjects (mean age 37.5 +/- 4.6) (group II). Initially, two dimensional echocardiography with color doppler examination, ambulatory Holter monitoring, multistage treadmill exercise test, transoesophageal pacing (TEP), recording of ASAECG and SAECG were made. Subsequently, pharmacologic autonomic system blockade was done by using intravenously propranolol (0.1 mg/kg) and atropine (0.02 mg/kg). The following time-domain parameters of ASAECG were calculated: the root mean square voltage for the last 10, 20, 30 ms of filtered P wave (RMS 10, 20, 30) and time duration of filtered P-wave (PWD). Moreover, the following parameters of SAECG were analyzed: the root mean square voltage of the signals in the last 40 and 50 ms of the filtered QRS (RMS 40, 50), the total time duration of QRS(t-QRS) and the duration of low amplitude signals < 40 microV (LPD). The autonomic blockade did not change significantly the obtained values of RMS 10, 20, 30 in subjects with MVP as follows: RMS 10 from 5.5 +/- 2.9 microV to 6.3 +/- 2.4 microV, RMS 20 from 8 +/- 3.7 microV to 8.5 +/- 2.4 microV and RMS30 from 10.2 +/- 4.9 microV to 10.3 +/- 3.3 microV. On the other hand, also the values of RMS 10 did not change statistically significant after blockade in control group (from 5.8 +/- 2.2 microV to 5.6 +/- 1.7 microV, p > 0.05), but the values of RMS20 i RMS30 increased significantly in this group, respectively from 7 +/- 3.1 microV to 8.3 +/- 3 microV (p < 0.02) and from 8.4 +/- 3.1 microV to 10.1 +/- 3.6 microV, (p < 0.05). The values of PWD have changed differently in the examined groups being under the influence of the autonomic blockade: in group I decreased from 119 +/- 12 ms to 114 +/- 28 ms (p > 0.05), whereas in group II increased from 110 +/- 13 ms to 116 +/- 9 ms (p < 0.02).(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 7479226 TI - [Frequency, diagnostic possibilities and importance of ischemic heart disease in patients with chronic obstructive pulmonary disease]. PMID- 7479227 TI - [Level of atrial natriuretic peptide (ANP) in plasma of patients with chronic renal failure during hemodialysis]. AB - Two groups of subjects (uraemic and control) were studied. The group A consisted of 20 patients treated by HD (haemodialysis). The mean age was x +/- SD--36.6 +/- 7.3 years, duration of haemodialysis treatment 32.8 +/- 7.7 months, cuprophan dialyzers and acetate containing solution--35 mEq/l--were used, time of HD--4 hours 3 times weekly, predialysis serum creatinine was 876 +/- 189 mumol/l (9.8 +/- 2.1 mg%). This group of patients was subdivided in two groups. The group I comprised 9 patients with hypotensive episodes that occurred during HD (systolic blood pressure < 90 mmHg), and the group II comprised 11 patients without hypotensive episodes. The control group comprised 20 healthy subjects (mean age 36.7 +/- 12.1 years, serum creatinine level 77 +/- 16 mumol/l (0.9 +/- 0.2 mg%). In all examined subjects the following experimental protocol was used. Blood pressure (BP) was determined at about 8 a.m. after an overnight rest. Then blood samples were withdrawn for estimation of ANP, haematocrit value (Ht), haemoglobin (Hb) and creatinine concentrations. Between 8 and 12 a.m. all examined uraemic subjects were dialyzed. After each hour of dialysis BP was measured and blood samples were taken. ANP (Peninsula Lab.Kids.) was measured using RIA method, and other biochemical parameters using routine methods. Serum creatinine and plasma ANP levels significantly decreased after HD. No significant differences were seen between the both uraemic groups. No significant correlations between systolic blood pressure, ANP level in examined group were observed. CONCLUSIONS: 1. In all uraemic subjects, plasma ANP level was significantly higher than in control subjects.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7479228 TI - [Plasma activity of diamine oxidase after heparin in patients with diabetic enteropathy]. AB - Diamine oxidase (DAO) is an enzyme found almost exclusively in the enterocyte of the villous tips. In the basal state plasma DAO activity is low but increases significantly after i.v. injection of heparin. It is well recognised that postheparin plasma activity of DAO reflects clearly the integrity of the small bowel mucosa. The aim of this study was to investigate postheparin plasma DAO activity in diabetics with enteropathy. The study was carried out in a group of 72 individuals subdivided into five groups. Group I consisting of 16 patients with type 1 diabetes, group II--20 patients with type 2 diabetes, group III--8 patients with newly diagnosed type 1 diabetes, group IV--8 diabetics with enteropathy and group V--the control group. Blood samples were taken at the time 0' and at 60' and 120' intervals following i.v. injection of heparin. DAO activity was assessed by RIA methods. Basal values of DAO were nearly the same in all of the subjects (< 50 pmol/min/ml). After injection of heparin a very high DAO activity was observed in all groups (> 400 pmol/min/ml) except group IV (< 270 pmol/min/ml) (p < 0.001). There were no significant differences between the groups according to the type of diabetes, its duration and the methods of treatment. In conclusion, the postheparin plasma DAO activity in diabetics with enteropathy was very low, but of itself, diabetes and its treatments had no effect on the results under investigation. PMID- 7479229 TI - [Chronic inflammation of the gastric mucosa and some immunologic aspects of Helicobacter pylori infection]. AB - The aim of the study was to determine what is the correlation between immunological phenomena and pathological changes in chronic gastritis and how it refers to the histology and endoscopy. PATIENTS AND METHODS: 42 patients with dyspepsia underwent following procedure: 1) gastroscopy and antral biopsy of 3 specimens; 2) histology; 3) immunofluorescence of the specimens in purpose to detect bound immunoglobulins using antibodies anti-human IgA + IgG + IgM labelled with rhodamine; 4) serologic test for anti-H.pylori antibodies. The research included 17 females and 25 males (ages 30-86, median 53.4 +/- 15.47). The obtained data were compared referring to mutual correlations and presented according to the kind of pathological changes depending on:--presence or absence of Helicobacter pylori infection;--presence or absence of anti-Helicobacter pylori antibodies;--presence or absence of lymphocytic infiltration;--presence or absence of bound immunoglobulins in gastric mucosa. RESULTS: --We have not observed ulcerations in anti-Helicobacter pylori seronegative group;--intestinal metaplasia and gastric ulcer were more frequent in bound immunoglobulins positive group;--biliary reflux was observed less frequently in lymphocyte infiltration negative group then in the positive one. CONCLUSION: pathological changes in chronic gastritis may depend not only on the presence of Helicobacter pylori infection, but also on the presence and quality of immunological response on its antigens. PMID- 7479230 TI - [Hypothermia during chemotherapy for lymphomas]. AB - There are only few reports in the literature on the occurrence of hypothermia after chemotherapy. It occurred after various cytostatics and lasted for few hours to several days. Our material consisted of 11 patients with malignant lymphoma who were given chemotherapy protocols including cisplatin. In 5 patients (2 with Hodgkin's disease and 3 with non-Hodgkin's lymphoma) who had high fever, after treatment the temperature decreased down to 34.3 degrees C. Hypothermia disappeared spontaneously after few days. The drug responsible for this effect in our patients was cisplatin. PMID- 7479231 TI - [Comparison of treatment effects with an angiotensin converting enzyme inhibitor- lisinopril and a calcium blocker--nifedipine retard on urinary albumin excretion in patients with non-complicated essential hypertension]. AB - The purpose of the study was to compare the effect of treatment with an angiotensin converting enzyme inhibitor (Lisinopril, MSD) or calcium blocker (Nifedipine retard, MSD) treatment during three months on blood pressure (measured with sphygmomanometric method and ambulatory blood pressure monitoring- ABPM) and urinary albumin excretion in essential hypertension class I acc. to WHO. Fifteen untreated patients aged 38 +/- 5 years with essential hypertension participated in the study and received diet with normal sodium content. Urinary albumin excretion was measured by RIA method in two 24 hour urine collections and mean value was calculated. ABPM was measured with Spacelabs monitor. After first examination 8 patients were randomly selected for the treatment with lisinopril and 7 patients to the treatment with nifedipine. The doses of both drugs were gradually adjusted to reach diastolic blood pressure below 90 mmHg. After 3 months of treatment urinary albumin excretion and blood pressure was found in both after treatment in patients treated with lisinopril but not in those receiving nifedipine. In patients treated with lisinopril a correlation between the decrease in systolic and diastolic blood pressure (measured by ABPM) and decrease of urinary albumin excretion was demonstrated. It was concluded that the normalization of blood pressure induced by lisinopril treatment in patients with uncomplicated essential hypertension and normoalbuminuria is accompanied with significant diminution of urinary albumin excretion which suggests preventive action of the drug in the development of microalbuminuria. Diminution of urinary albumin excretion caused by lisinopril is probably due to both the decrease of blood pressure and the specific renal action of the drug. PMID- 7479232 TI - [Use of fuzzy set theory in diagnostics]. PMID- 7479234 TI - [Langerhans cell histiocytosis--immunologic and clinical aspects]. PMID- 7479233 TI - [Use of fuzzy set theory in therapy]. PMID- 7479235 TI - [Extraintestinal salmonellosis in patients with blood diseases caused by Salmonella enteritidis]. AB - Four cases of extraintestinal salmonellosis caused by Salmonella enteritidis were described. Underlying diseases in the three patients were haematological neoplasms (2 splenectomized and 1 with massive leukemic infiltrations of the spleen) and in the fourth haemophilia B: only that patient had a prior symptomatic intestinal infection. Blood cultures for S.ent. were positive in all patients and additionally in that suffering from Hodgkin's disease urine, lymph node and stool cultures also showed S.ent. In the haemophiliac patient culture of suppurated hematoma was positive. All patients recovered from S. sepsis but three then died of their neoplastic diseases. The haemophiliac patient is in a good condition. Various disturbances in the immunological tests were observed. PMID- 7479236 TI - [Langerhans Cell Histiocytosis in adults]. AB - Five cases of Langerhans' cell histiocytosis in adult patients were reviewed. Signs, symptoms, the course of the disease and methods of treatment were presented. Results of tests, including results of organ biopsies that provided definitive diagnosis in most cases, were also analysed. PMID- 7479238 TI - [Role of prostaglandins F and E in regulation of bronchial activity]. AB - Prostaglandins (PGs) belong to the family of eicosanoids. A main substrate in their biosynthesis is arachidonic acid. Involvement of PGs in regulation of bronchial activity is a complex process and far from being fully understood. In 1968 it was shown that prostaglandins E1 and E2 cause bronchodilation, while prostaglandin F2a constricts an isolated human smooth muscle. The recent studies which revealed protective effects of PGE2 on bronchial reaction induced in patients by ultrasonically nebulized distilled water, allergen, metabisulfite, or exercise, revitalized interest among clinicians. Reports that PGE2 inhibits airways response in bronchial provocation with lysine aspirin, allow to believe that PGE2 plays a role in pathogenesis of aspirin induced asthma. PMID- 7479239 TI - [Treatment strategy for rheumatoid arthritis]. PMID- 7479237 TI - [Unusual clinical course of ruptured aortic aneurysms--reports of three cases]. AB - Three cases of unexpected clinical course of ruptured aortic aneurysm have been presented in patients of their 7th decade life. All of them had arterial hypertension. Signs and symptoms on admission to hospital (dysphagia, chest and interscapular pain, hematemesis, abdominal pain, elevated body temperature and diminished exercise tolerance) were non-specific of aortic aneurysm, suggesting other disease. Dramatic clinical course with hypovolemic shock in two cases led to death. One of them refused surgery. In the third one, in spite of blood effusions to pleural cavity, pericardial sac and mediastinum, effective hypotensive therapy with a preservation of the slow heart rate and fluid evacuation from pericardial sac, gave the opportunity to perform elective surgery. Aortic dissection often presents an atypical course and when suspected, all available imaging technics including computed tomography and nuclear magnetic resonance must be used. PMID- 7479240 TI - [Increased cholesterol content in erythrocyte membranes of patients with chronic renal failure treated with hemodialysis]. AB - The investigate whether the blood rheological disturbances occurring in hemodialyzed patients are related to the erythrocyte lipid composition, cholesterol and phospholipid content was studied of the erythrocyte membrane. The study included 40 patients with chronic renal insufficiency. 34 of them had hypertension and proteinuria. The cholesterol/phospholipid ratio was significantly higher in the patients after hemodialysis (mean 2.48; SD 0.14) compared with the patients before hemodialysis (mean 2.08; SD 0.19). There was a significant increase of cholesterol in subgroup with hypertension and proteinuria. A significant correlation was found between membrane cholesterol/phospholipid ratio and serum levels of LDL-cholesterol. PMID- 7479241 TI - [levels of erythropoietin in serum of patients with primary hyperparathyroidism]. AB - The present study aimed assess the relationship between erythropoietin (EPO) and parathormone (PTH) secretion in patients with primary hyperparathyroidism (PHP). Seventeen patients with PHP and 24 patients with uncomplicated cholelithiasis were examined before and during 14 days after surgical treatment. In PHP patients plasma EPO levels were significantly higher than in patients with cholelithiasis (31.5 + 4.3 vs 14.1 + 0.9 mU/ml, p < 0.01). After cholecystectomy transient increase of plasma EPO level was noticed postoperatively during the first 6 days. Such an increase of plasma EPO was absent in PHP patients after removal of the cause of primary hyperparathyroidism. Only in PHP patients a significant negative correlation was found preoperatively between plasma EPO level and Hct value or creatinine clearance respectively. CONCLUSIONS: 1. Patients with PHP are characterized by significantly elevated plasma EPO levels which are related to the degree of anaemia and decrease of GFR. 2. In patients with PHP presence of significant relationship between EPO and PTH secretion could not be proven. PMID- 7479242 TI - [The diagnostic value of cytology in children on continuous ambulatory peritoneal dialysis (CAPD)]. AB - Peritonitis is a common clinical problem that occurs in patients undergoing CAPD. Early diagnosis of this complication is difficult in some cases and elevated peritoneal cell count may be a misleading factor in diagnosis. The aim of the study was to determine the cell count in consecutive dialysate exchanges after starting CAPD. Elevated peritoneal cell count was found in 17 of 21 (80%) analyzed periods at the beginning of CAPD. Peritonitis was diagnosed in 5 of these patients and confirmed by culture in 4 and clinical observation in 1. In the remaining 12 cases (70%) the elevated cell count normalized during consecutive exchanges of dialysate. The coexistence of such nonspecific symptoms as fever, abdominal pain of varying intensity, cloudy fluid with or without the presence of fibrin can be the cause of diagnostic mistake and institution of unnecessary antibiotic treatment. In doubtful case, the reduction of the dwell time to 2 hours and observation of the cell count in consecutive exchanges is very helpful, especially if rapid culture diagnosis is not available. PMID- 7479243 TI - [Doppler examination of portal venous flow in liver neoplasms]. AB - An attempt was made to measure blood flow of portal system at various stages of neoplastic changes in the liver, which could be of some prognostic and therapeutic value. In the group of 80 studied persons, 20 patients had hepatic angioma, 6 patients hepatic cancer, 24 patients metastatic cancer affecting to the liver. Control group consisted of 50 healthy volunteers. No disturbances of blood flow in portal system in the group of patients with hepatic angioma were found. In the group with primary hepatic cancer decrease of TAV and TF in portal and splenic veins were noted, especially in 2 cases with numerous changes with relatively big sum of diameters, as compared with control method. In group with metastases of cancer to the liver slight decrease of blood flow in portal and splenic veins was observed as compared to the control group. Decrease in blood flow in the portal system was seen in cases with numerous and relatively big neoplastic focuses in the liver. Blood flow monitoring might be helpful in evaluation of the progress or regression of infiltrative changes in the liver. PMID- 7479245 TI - [Clinical picture and outcome of conservative treatment for obstructive sleep apnea]. AB - The aim of the study was to assess the clinical picture of patients with obstructive sleep apnea (OSA) and to investigate the long term effectiveness of treatment with continuous positive airway pressure (CPAP). 170 subjects were investigated with Mesam 4 screening device. Full polysomnography was performed in suspected patients confirming the diagnosis (AHI > 10) in 72 cases (69M, 3F), mean age 46.7 +/- 8.4 yrs, mean weight 110 +/- 19 kg. Snoring (100%) apneas (96%), excessive daytime sleepiness (98%) and nocturia (73%) were the most common symptoms. Application of CPAP therapy resulted in elimination of apneas, improvement in sleep architecture and in arterial blood oxygen saturation. Apnea index decreased from 63 +/- 22 to 4 +/- 4. First night compliance to CPAP was about 95%. OSA symptoms rapidly regressed. Follow-up examination was performed in 45 patients who used CPAP for more than 1 year. All subjects presented with improvement in quality of life, all but one were using CPAP almost every night (mean 6.6 +/- 0.9 night per week). The most common side effects of CPAP were uncomfortable nose mask (42%), throat dryness (27%) and eye irritation (24%) We concluded that CPAP treatment in OSA occurs to be effective, long term compliance is high, side effects are benign and mostly due to poorly fitting mask. PMID- 7479244 TI - [Evaluation of early treatment results in Hodgkin's disease during the cytostatic protocol containing methotrexate]. AB - 28 patients suffering from advanced Hodgkin's disease were treated according to protocol containing 7 cytostatic drugs including methotrexate and corticosteroids, but not cyclophosphamide, chlormethine or procarbazine. 22 patients i.e. 78.5% attained complete remission and 5 i.e. 17.8% a partial one. Drug tolerance was satisfactory: out of side-effects the most common was myelosuppression, especially granulocytopenia. Our early results were similar to those obtained with MOPP-like protocols, but early undesirable effects less often and of a smaller degree. Because the protocol has been aimed at avoiding late toxicity of cytostatics, its final evaluation will be possible in the future. PMID- 7479246 TI - [Endocrine abnormalities in active nephrolithiasis]. PMID- 7479247 TI - [Autoimmune hypoglycemia]. PMID- 7479248 TI - [Pancreatic ascites]. AB - Two cases of pancreatic ascites have been presented. 1. A 29 year old man, heavy alcoholic, with fast growing ascites presented since long biochemic features of pancreatitis. The diagnosis of the pancreatic origin of ascites was made on the ground of the high level of amylase in the ascitic fluid. Conservative treatment was effectless. The patients refused surgical therapy. Then he died in septic shock. Autopsy confirmed the pancreatic origin of ascites. 2. A 43 year old man heavy alcoholic as well admitted because of fast growing ascites. As in the first case examination of the fluid confirmed the diagnosis of pancreatic origin of ascites. Again a high level of amylase and protein was found. After clinical treatment and parenteral nutrition the condition of the patient improved and he was dismissed 4 weeks later. PMID- 7479249 TI - [Implications of chaos theory in pharmacotherapy]. PMID- 7479250 TI - [Differences and similarities between asthma and chronic obstructive pulmonary diseases]. PMID- 7479251 TI - [Activity of pyrimidine 5'nucleotidase (py5'Nd) of erythrocytes in essential hypertension]. AB - Activity of pyrimidine 5'nucleotidase was measured in hemolysate of erythrocytes of healthy persons and patients with essential hypertension. Cytidine 5'monophosphate (CMP) and uridine 5'monophosphate (UMP) were used as the substrates for evaluation of activity of so called I-isoenzyme and uridine 3'monophosphate (U3'MP) was used as a substrate for the II isoenzyme of Py5'Nd. It was found that the activity of Py5'Nd I was lower in hypertensives (26.8 mU/gHb (CMP)) and 69.3 mU/gHb (UMP) in comparison with normotensives (62.3 and 117.4 mU/gHb respectively) (p < 0.05). The activity of Py5'Nd II did not differ between studied groups. Possible metabolic consequences of decreased activity of Py5'Nd are discussed. PMID- 7479252 TI - [Interdigestive gallbladder volume and and meal-induced gallbladder emptying in patients with diabetes]. AB - The aim of the study was to find whether the type of diabetes and the duration of this disease causes any influence on gallbladder (GB) emptying, after exclusion of possible effect of obesity and a hyperglycaemia over 7 mmol/l (126 mg%) on this phenomenon. The investigations were carried out in 44 subjects with diabetes mellitus, among them 22 with type I and 22 with type II. These groups were divided according to disease duration: shorter or longer than 3 years. Two control groups (CI, CII) comprised each 20 healthy subjects. The age of CI and CII corresponded to the age of diabetes groups type I and II, respectively. All subgroups as well as the control groups were matched according to sex percentage structure, body mass index (BMI) and body surface (BS). In groups with type I and II diabetes, the number of cases with diabetic neuropathy was comparable. There was not found any significant (p > 0.05) influence of diabetes on the interdigestive GB volume, neither non-standardized nor standardized with regard to standard BS (1.73 m2). In comparison to the control groups, after a test meal, there was in all diabetic groups a statistically significant (0.001 > p < 0.05) reduction of GB emptying observed. This diminution was most expressed in the group with long-lasting type I diabetes. PMID- 7479253 TI - [Changes of transport functions in peritoneum caused by glucose and its penetration in vitro]. AB - The changes of the transperitoneal molecules transport caused by glucose and dynamics of its transport have been determined in the case of undamaged tissue and the membrane with injured mesothelium in the in vitro studies. In the intact glucose induced increase of peritoneal uric acid absorption (40%) and lowering of urea and gentamicin excretion (25-40%). Glucose in the dialysis fluid lowered uric acid absorption and urea excretion (20-40%) in denuded animal peritoneum. The transperitoneal glucose transport remained constant at 2.61 x 10(-4) x cm x s 1. The mesothelium destruction caused an increase (32%) of mean values of absorption and excretion of this compound. The obtained results suggest that modifications of transport functions of the peritoneum caused by glucose depend rather on direct action of hypertonicity and metabolic effects of glucose than of the osmotic gradient. Furthermore, these changes may contribute to the rapid transperitoneal transport of glucose which increase after the destruction of the mesothelium. PMID- 7479254 TI - [Activity of converting enzyme angiotensin I to angiotensin II (ACE) in patients with diabetes type II]. AB - The aim of the work was evaluation of activity of angiotensin converting enzyme (ACE) in non-insulin dependent diabetes mellitus depending on evidence of hypertonia, duration of diabetes and possibility of fat metabolism disturbances. 20 patients were studied (16 women and 4 men, average age 57.5 years). Average time of duration of diabetes was approximately 9 years (from 1 year to 25 years). The control group included 68 people (34 women and 34 men) in age of 19 to 67 years. Convertase activity was determined by Friedland and Silverstein's spectrofluorometric assay. Average ACE activity values in diabetic patient (44.45 +/- 13.14 mmol/ml/min) were close up to control group values and were not significantly different. Similar significant difference was not ascertain between average ACE activity values in patients with more than and less than 10 years of diabetes, in patients with hypercholesterolemia over and less 300 mg%, in patients with and without hypertonia and in patients with and without LDL hypercholesterolemia over and less 200 mg%. Positive correlation was observed between ACE activity values and triglyceride levels in patients with less than 10 years of diabetes (r = 0.88, p < 0.01) and in patients with hypercholesterolemia over 300 mg% (r = 0.75, p < 0.01). In this last group, in addition, the positive correlation was present between ACE activity values and total cholesterol levels (r = 0.86, p < 0.001). Correlation between ACE activity and total cholesterol level was ascertained in patients with LDL hypercholesterolemia over 200 mg% too (r = 0.81, p < 0.01).(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7479255 TI - [Clinical course of Waldenstrom macroglobulinemia based on personal observation of 24 patients]. PMID- 7479256 TI - [Infection in the dialysis unit as a contributing factor to the problem of hospital acquired infections]. AB - Hospital acquired infections are especially dangerous for patients with chronic renal insufficiency. An epidemic of hospital acquired infection of dramatic course in described in 13 patients (8 being on haemodialysis (HD) and 5 treated conservatively), with 15 episodes of septic syndrome (12 cases) or septic shock (3 cases). Initial cultures of blood, urine and pharyngeal swabs were positive only in 5 patients treated conservatively. Initial cultures of dialysing fluid from all 12 monitors of HD in our Dialysis Unit revealed the presence of Gram negative bacteria (Achromobacter) in blood of 3 patients as well as in the dialysing fluid compartments from 2 monitors of Gambro AK 10/100. The possible role of Gram-negative, apparently benign water rods, in the pathogenesis of hospital acquired opportunistic infections in Dialysis Unit were discussed. PMID- 7479257 TI - [Some aspects of epidemiology in insulin-dependent diabetes mellitus (IDDM) (I)]. AB - Epidemiological studies based on registries from many centers in the world revealed geographical variation of IDDM, with the highest incidence in Scandinavian countries. There is a gradient north to south in occurrence of IDDM, and differences in incidence between urban and rural populations. Poland is a country with a rather low incidence of IDDM, not exceeding 6/100,000 in the population 0-29 years of age until 1992. Generally a higher incidence of IDDM has been observed in Whites, there are differences between migrating populations ethnically homogenous. The highest incidence occurs in 10-14 age group, with male sex predominance. PMID- 7479258 TI - [Some aspects of epidemiology in insulin-dependent diabetes mellitus (IDDM) (II)]. AB - Seasonality is a characteristic feature of IDDM incidence. The lowest incidence is observed in summer, whereas the highest in winter/autumn. The risk of IDDM increases in children with independently: breast feeding shorter then 3 months, feeding with cow's milk instead of breast feeding, early introducing of solid diet in babies, or high consumption of cow's milk in later years. Children with new onset IDDM have elevated levels of the antibodies against cow's milk proteins and beta-lactoglobulin. It has been proved that there is a connection with HLA DQalfaArg52 i DQbeta-Asp57 and polymorphic region of insulin gene, and the IDDM susceptibility. PMID- 7479259 TI - [Importance of nitric oxide in physiological states and pathology]. PMID- 7479260 TI - [Overdose of methotrexate with a fatal outcome in a patient with rheumatoid arthritis]. AB - A 52-year old female patient with rheumatoid arthritis (RA) had mistakenly taken 112.5 mg methotrexate (MTX) over a period of five days. As a result, extensive erosions along with necrotic changes occurred in the oral cavity, the groins and vulva mucous membrane. Haemorrhages in the gastrointestinal tract, dermal xanthochromia, interstitial pneumonia and progressive renal insufficiency, as well as hair loss, also followed. Laboratory examination showed peripheral blood agranulocytosis, a decrease in the overall number of blood platelets, anaemia, an increase in the level of transaminases and bilirubin, along with megakaryocyte deficiency. Cultivated dermal specimens initially developed bacteria, then Candida albicans. The patient died on the 22nd day after the overdose from pneumonia and extensive mycosis. PMID- 7479261 TI - [Secondary myelodysplastic syndromes]. PMID- 7479262 TI - [Treatment of disseminated intravascular coagulation syndrome]. PMID- 7479263 TI - [Acute, post-transfusion lung injury (TRALI) and related states]. PMID- 7479264 TI - [Principles of procedure for patients treated with cytostatics]. PMID- 7479265 TI - [Acute agranulocytosis--pathogenesis, diagnosis and treatment]. PMID- 7479266 TI - [Preparing patients with hematologic diseases for surgical interventions]. PMID- 7479267 TI - [Acute hemolysis--diagnosis and management]. PMID- 7479268 TI - [Essential thrombocythemia]. PMID- 7479269 TI - [Possibilities for inhibiting the progression of chronic renal diseases]. PMID- 7479270 TI - [Gastrointestinal non-Hodgkin's lymphomas. Personal observations]. AB - The localization, symptoms, endoscopic findings, histologic type, infiltration of bone marrow, lymphoma cell presence in the blood, and results of therapy were evaluated in 18 patients with gastrointestinal lymphoma. Only 9 patients met the criteria of primary gastrointestinal lymphoma, the remaining 9 presenting an extra-nodular localization of systemic disease, i.e. secondary lymphoma. The most common site of process was stomach, and most of the patients had histologically documented a high grade malignant lymphoma. Necessity of supplementing surgery with chemo- and/or radiotherapy was discussed. Such combined treatment should give a 10-year survival time in 60% of the treated patients (confirmed with the Kaplan-Meier's method). The authors concluded that stomach is the most common localization of gastrointestinal lymphoma, and that gastrointestinal non Hodgkin's lymphoma diagnosis should include peripheral blood and bone marrow haematological examinations which clearly rise the reliability of differentiation between primary or secondary lymphomas. PMID- 7479271 TI - [The influence of HIV infection on development of tuberculosis]. PMID- 7479273 TI - [Megakaryocytes as a component of bone marrow-blood barrier]. PMID- 7479272 TI - [Familial occurrence of pheochromocytoma]. AB - Familial occurrence of pheochromocytoma in mother and two sons (11 and 14 years old) is described. The diagnosis was based on biochemical tests and localization methods. Bilateral tumors were demonstrated on computerized tomographic scanning in two of our patients. No association with multiple endocrine neoplasma was found. Removal of the tumors resulted in normalization of blood pressure and biochemical parameters. The histopathological examination of the tumor confirmed the diagnosis. A possibility of familial occurrence should be considered in patients with pheochromocytoma and appropriate diagnostic tests must be performed in their near relatives. PMID- 7479274 TI - Fumes from the spleen. PMID- 7479275 TI - The Tasmanian SIDS Case-Control Study: univariable and multivariable risk factor analysis. AB - A population-based retrospective case-control study has been conducted in Tasmania since October 1988. Study measurements pertained to the scene of death of last sleep, as well as a verbal questionnaire on relevant exposures. From 1 October 1988 to 1 October 1991, 62 cases of sudden infant death syndrome (SIDS) occurred. Case response rate for retrospective interviews was 94% (58/62). The initial control response rate was 84% (101/121). After stratification for maternal age and birthweight, there was no increase in risk associated with the usual side position (odds ratio [OR] 1.05 [0.27, 5.02]), compared with the supine position (OR 1.00, reference). The prone position was associated with increased risk [OR 5.70 (1.67, 25.58)], relative to the supine position. In the final multivariable model, predictors of SIDS in this study were usual prone position (P < 0.001), maternal smoking (P = 0.008), a family history of asthma (P = 0.045) and bedroom heating during last sleep (P = 0.039). Protective factors were maternal age over 25 years (P = 0.013) and more than one child health clinic attendance (P = 0.003). The results provide further support for current health education activities which aim to inform parents of modifiable risk factors for SIDS, including the prone sleeping position, thermal stress and infant exposure to tobacco smoke. PMID- 7479277 TI - Racial differences in the patterns of preterm delivery in central North Carolina, USA. AB - In order to assess racial differences in rates of idiopathic preterm labour, preterm premature rupture of membranes, and medically indicated preterm delivery, the authors analysed data on 388 preterm (< 37 completed weeks of gestation) births (7.9% of all births) occurring between 1 September 1988 and 31 August 1989, in three central North Carolina counties. The crude relative risk (RR) of preterm birth among black women compared with white women was 2.6 [95% confidence interval (CI) 2.1, 3.1]. With adjustment for age, gravidity, marital status, education, and county of residence, the estimated relative risk for black women compared with white women was 2.1 (95% CI 1.1, 4.1) for medically indicated preterm delivery, 1.6 (95% CI 1.1, 2.3) for preterm birth as a result of preterm labour, and 1.9 (95% CI 1.2, 3.1) for preterm premature rupture of membranes. Compared with white women, black women were at the highest risk of a preterm birth before 34 weeks of gestation (RR = 2.9; 95% CI 1.8, 4.7). The risk of medically indicated preterm delivery at 36 weeks was considerably higher for black women than for white women (RR = 3.4; 95% CI 1.1, 10.2). For a better understanding and ultimately a reduction of the risk for preterm delivery among black women, investigation of specific aetiological pathways and gestational age groups may be required. PMID- 7479276 TI - Birthweight, preterm births and neonatal mortality in Belgium and the United States. AB - Belgium is known to have a lesser low birthweight rate and a lower infant mortality rate than the United States. We used previously unpublished data to show that beneath this comparison lies a more complicated picture. Singleton live birth certificates for 1986-87 were analysed. Despite a lower mean birthweight in Belgium (3360 g) than in the United States (3420 g), Belgium had fewer (4.9%) low birthweight infants than the US (5.9%) because of fewer preterm births (4.4 vs. 9.3%). Consistent with the excess of preterm births in the US, the residual distribution of birthweight was smaller in Belgium (2.2% vs. 3.1%). Whereas neonatal mortality was 4.8/1000 in Belgium and 5.6/1000 in the US, birthweight specific neonatal mortality was higher in Belgium. The challenge for Belgium is to improve the survival of newborns regardless of their birthweight. In the US, the task is to eliminate the excess of small preterm infants. PMID- 7479278 TI - Did preterm deliveries continue to decrease in France in the 1980s? AB - To study the trends in gestational age and birthweight in France, we compared data from three surveys that were based on representative samples of births in 10 French regions and were conducted in 1972, 1981 and 1988-89. Only single livebirths were considered in the analysis. The rate of preterm births decreased from 7.9% in 1972 to 5.8% in 1981 and 4.0% in 1988-89. The corresponding rates of children with a birthweight below 2500 g were 5.4, 4.3 and 4.7%. The mean birthweight of preterm babies born in 1988-89 was lower than the mean birthweight of those born in 1972 and 1981. The reduction in preterm births cannot be attributed to the improved accuracy of gestational age determination, but the general use of ultrasound may have played a major role in the apparent reduction of the mean birthweight of preterm babies. PMID- 7479279 TI - Do pregnant smokers eat differently from pregnant non-smokers? AB - This article describes a study of the relationship between diet and smoking in a group of 821 Norwegian pregnant women. The study is part of a multi-centre project, examining risk factors for intrauterine growth retardation. Two 3-day dietary records were collected during the 17th and 33rd week of pregnancy. Information on smoking habits and other relevant parameters were collected through an extensive questionnaire. The results showed that the smokers consumed significantly less than the non-smokers of bread, cakes and cookies, vegetables, fruits and berries, cheese, yoghurt, low fat milk, juice and tea. The smokers also consumed significantly more meat, margarine, whole milk, soft drinks and coffee than the non-smokers on both occasions. The diet of the smokers contained significantly less protein, carbohydrate, dietary fibre, thiamin, riboflavin, vitamin C, calcium and iron as compared with the non-smokers. Fat contributed significantly more to the energy content of the diet of the smokers and it is concluded that their diet was less nutritious than that of the non-smokers throughout pregnancy. PMID- 7479280 TI - Maternal Lyme disease and congenital malformations: a cord blood serosurvey in endemic and control areas. AB - This report describes a cohort study of over 5000 infants and their mothers who participated in a cord blood serosurvey designed to examine the relationship between maternal exposure to Lyme disease and adverse pregnancy outcome. Based on serology and reported clinical history, mothers of infants in an endemic hospital cohort are 5 to 20 times more likely to have been exposed to B. burgdorferi as compared with mothers of infants in a control hospital cohort. The incidence of total congenital malformations was not significantly different in the endemic cohort compared with the control cohort, but the rate of cardiac malformations was significantly higher in the endemic cohort [odds ratio (OR) 2.40; 95% confidence interval (CI) 1.25, 4.59] and the frequencies of certain minor malformations (haemangiomas, polydactyly, and hydrocele), were significantly increased in the control group. Demographic variations could only account for differences in the frequency of polydactyly. Within the endemic cohort, there were no differences in the rate of major or minor malformations or mean birthweight by category of possible maternal exposure to Lyme disease or cord blood serology. The disparity between observations at the population and individual levels requires further investigation. The absence of association at the individual level in the endemic area could be because of the small number of women who were actually exposed either in terms of serology or clinical history. The reason for the findings at the population level is not known but could be because of artifact or population differences. PMID- 7479281 TI - Epidemiology of Down's syndrome in a Scottish city. AB - The aim of the study was to investigate aspects of the epidemiology of Down's syndrome (DS) in a population subjected to continuous surveillance. Data were obtained from the Glasgow Register of Congenital Anomalies. Over the period 1980 90, 173 cases of DS were identified (1.2 per 1000 births), of which 135 (78.1%) were livebirths, five (2.9%) were stillbirths and 33 (19.1%) were terminations following prenatal diagnosis. The increasing risk of DS with advancing maternal age was confirmed, although most DS pregnancies occurred in women aged under 35. While the birth prevalence of DS did not vary significantly over time, there was a significant increase in DS pregnancy prevalence. The proportion of all DS pregnancies terminated showed a statistically significant rise from 14% in 1980 87 to 31% in 1988-90 following the introduction of a new screening marker for DS risk. Almost half of DS pregnancies in women aged 35 and over ended in termination following prenatal diagnosis compared with only 7% in women under 35. There was a statistically significant excess risk of DS pregnancy in the more prosperous areas of the city; this excess persisted, but was not statistically significant, after standardising the rates for maternal age. These findings suggest that there is no scope for reducing service provision for DS children on the basis of the epidemiological trends, that the antenatal screening programme, while benefiting from the recent introduction of serum markers of risk, continues to have a greater impact on older women, and that further aetiological research using population based registries is required. PMID- 7479282 TI - Efficiency of sampling: birthweight and gestational age distributions in two cohorts, < 31 weeks and 500-1499 grams. AB - We studied the efficiency of two common sampling strategies used to assemble cohorts to study the long-term problems of preterm infants: infants with birthweights of 500-1499 g, and infants with gestational ages (GA) of < 31 weeks. Birthweight, GA and 2-year outcome data from a population based study of infants < 2001 g, the Central New Jersey Brain Hemorrhage Study (NBH), were used to define the birthweight and GA distributions, at enrollment and at the age of 2 years, of overlapping subsets: infants 500-1499 g (n = 599) and infants < 31 weeks of age (n = 522). Using frequencies from the NBH study, we estimated that 1000 infants of 500-1499 g enrolled at birth would produce 712 infants at the age of 2 years, 498 below 31 weeks and 214 above. Enrolling 1000 infants < 31 weeks would produce a cohort of 697 infants at the age of 2, all of whom were < 31 weeks. Neither sampling strategy maximised the statistical power to investigate the pathophysiological determinants of long-term outcomes associated with short GA. Both methods oversampled older GAs. A stratified sampling technique based on GA, designed to produce equal numbers of subjects at each week of GA, would improve statistical power to study long-term outcomes. As we move from descriptive to analytical studies of preterm infants, we need to devise efficient, GA-based, sampling strategies that maximise statistical power to test pathophysiological hypotheses. PMID- 7479283 TI - Low birthweight as a determinant of infectious disease mortality: a methodological evaluation. AB - Low birthweight (LBW) is cited as an important determinant of infectious disease mortality during infancy, both in developed and developing countries. We applied methodological standards to evaluate the scientific validity and clinical applicability of epidemiological studies of this relationship. Three published studies, all reporting an elevation of the risk of infectious disease mortality in infants born with LBW, were identified. Noteworthy methodological deficiencies found in one or more studies included failure to obtain objective data about birthweight and gestational age for a high proportion of study subjects, lack of explicit criteria for assigning causes of death, failure to safeguard against biases in assessment of cause of death and inadequate control for potential confounding variables. In addition, clinical application of the results of these studies was limited by lack of exploration of the roles of preterm birth vs. small-for-gestational-age (SGA) birth, as well as by inattention to the role of congenital anomalies, in explaining the elevated risk. Although our evaluation does not necessarily negate the results of these studies, the failure of any of the studies to comply with all of the standards highlights the need for further research to provide credible evidence regarding the presence and strength of a relationship between LBW and infectious disease mortality in both developed and developing countries. PMID- 7479284 TI - Isolation and primary structures of neuropeptides of the AKH/RPCH family from various termite species. AB - We have isolated neuropeptides of the AKH/RPCH family from extracts of whole heads of four termite species (Mastotermes darwiniensis, Microhodotermes viator, Hodotermes mossambicus, and Trinervitermes trinervoides) using the effect of mobilizing lipids in Locusta migratoria for bioassay. Isolation was essentially achieved by two steps of reversed-phase chromatography (on phenyl-support followed by C-18). The peptides were identified by Edman degradation after deblocking with oxoprolyl peptidase. Each termite species contained only one AKH/RPCH family member. The primary structure in M. darwiniensis and T. trinervoides is pGlu-Val-Asn-Phe-Ser-Pro-Asn-Trp-NH2, a peptide previously found mainly in cockroaches and code named Pea-CAH-I. The peptide from M. viator has the primary sequence pGlu-Ile-Asn-Phe-Thr-Pro-Asn-Trp-NH2; it is a novel member of the family and is code-named Miv-CC (Microhodotermes viator corpus cardiacum peptide). Phylogenetic relations between the known cockroach and mantid AKH/RPCH octapeptides and the termite peptides from this study could be revealed employing the parsimony method. Based on a computer analysis, using PAUP 3.1.1., we concluded that termites are plesiomorphic with regard to cockroaches, and mantids are the sister taxon to the termite/cockroach group. PMID- 7479285 TI - Structure-activity relations of fulicin, a peptide containing a D-amino acid residue. AB - Structure-activity relations of fulicin (H-Phe-D-Asn-Glu-Phe-Val-NH2) isolated from the ganglia of a pulmonate mollusc Achatina fulica were investigated on the contraction of penis retractor muscle of this snail. The contraction-enhancing activity of fulicin was reduced to about 1/3000 when D-Asn2 was replaced by L Asn, whereas substitution of the other D-amino acid for D-Asn2 showed no significant reduction of the activity. On the contrary, if a D-amino acid was introduced into any position other than position 2 the activity was reduced remarkably. The presence of Glu3 and five amino acid residues as a whole were also required for activity. PMID- 7479286 TI - Effects of surgical denervation on substance P and calcitonin gene-related peptide in adjuvant arthritis. AB - The occurrence of substance P (SP) and calcitonin gene-related peptide (CGRP) was assessed in the ankle joints and dorsal root ganglia (L2-L6) by immunohistochemistry and radioimmunoassay (RIA) after unilateral sciatic nerve section in adjuvant arthritis. Nerve section in arthritic rats had no clear-cut effect on warmth, redness, and swelling of ankle joints, but significantly affected the occurrence of SP and CGRP. Immunohistochemical analysis showed an almost complete disappearance of SP-positive fibers in the ipsilateral ankle joint in normal rats after axotomy, whereas in arthritic rats occasional SP positive fibers remained. In dorsal root ganglia, only occasional SP-positive cells could be detected in normal and arthritic rats after axotomy. A similar but somewhat less pronounced effect of axotomy was noted for neuronal CGRP-LI. RIA showed a decrease in SP in ankle joints by 45% in normal rats and 58% in arthritic rats; the decrease in CGRP was 41% and 47%, respectively. In dorsal root ganglia, the decrease in SP after surgical denervation was 25% in normal rats and 54% in arthritic rats; the decrease in CGRP was 18% and 27%, respectively. The tissue concentrations of SP and CGRP in ipsilateral ankle joints and dorsal root ganglia were consistently correlated in normal as well as arthritic rats. The present study shows that an interruption of the nerve supply to joints cannot fully prevent the development of arthritis, although it significantly reduces the occurrence of sensory neuropeptides. PMID- 7479287 TI - Enhancement of NGF gene expression in rat brain by the memory-enhancing peptide AVP(4-8). AB - Northern blot analysis of nerve growth factor (NGF) was used to evaluate the effect of exogenous AVP(4-8) on the transcription of NGF gene in rat brain. NGF expression was found to be significantly enhanced by exogenous AVP(4-8) in the hippocampus as well as in the cerebral cortex in a time period of 12 h. This effect was inhibited by an antagonist to AVP(4-8). In addition, gel mobility shift assay was also used to observe the in vitro expression of c-fos gene in rat hippocampal slices. Our results suggest that NGF gene is one of the target genes responsible for memory-enhancing responses induced by AVP(4-8) and that the enhancement of NGF gene expression may share the signaling pathway mediated by AVP(4-8) receptor and c-fos gene expression. PMID- 7479288 TI - Involvement of septide-sensitive tachykinin receptors in inositol phospholipid hydrolysis in the rat urinary bladder. AB - The selective NK2 agonist [Lys5-MeLeu9,Nle10]NKA(4-10) markedly stimulated [3H]inositol monophosphate (PI1) formation in prisms from the rat urinary bladder. This response was blocked by the NK2 antagonist SR 48968. Senktide (NK3 agonist) was inactive. Septide, a short SP analogue, and the NK1 agonists [Pro9]SP and [Sar9,Met(O2)11]SP also stimulated [3H]IP1 formation and several NK1 tachykinin antagonists (RP 67580, CP 96345, GR 82334, and [D-Pro9,t beta BPr10,Trp11]SP) were more potent in blocking the septide than the [Pro9]SP response. GR 82334 was the most discriminative. SR 48968 (10(-6) M shifted the [Pro9]SP dose-response curve but did not modify the septide dose-response curve. Septide had a low affinity for [3H][Pro9]SP binding sites, suggesting further that septide and NK1 agonists act on different receptors. Finally, both [Pro9]SP and [Sar9,Met(O2)11]SP blocked the septide-evoked response, acting as partial agonists at the septide-sensitive tachykinin receptors. PMID- 7479289 TI - Growth hormone-releasing hormone mediates feeding-specific feedback to the suprachiasmatic circadian clock. AB - Growth hormone-releasing hormone (GHRH) is known to stimulate food intake in a circadian phase-dependent manner in rats. The suprachiasmatic nucleus (SCN) is an important site of action for this effect. In light of the central role played by the SCN in the control of circadian rhythms, together with the phase-dependent nature of GHRH-induced feeding, we sought to determine the possible involvement of SCN GHRH activity in the regulation of circadian rhythmicity. Two studies were conducted using hamsters as subjects. Study one replicated the daytime feeding stimulatory effects of GHRH in hamsters, thereby validating its appetitive effects in this species. Study two showed that, in free-running hamsters, intra SCN microinjections of GHRH produced phase advances when injected during the subjective day while having little effect during the subjective night. The GHRH phase-response curve was found to resemble that observed for nonphotic influences on the clock. It is suggested that GHRH input to the SCN is a neural representation of a nonphotic influence (perhaps feeding specific) on the clock. PMID- 7479290 TI - Dissociated feeding and hypothermic effects of neuropeptide Y in the paraventricular and perifornical hypothalamus. AB - The present study investigated the effects of neuropeptide Y (NPY) on food intake and body temperature (Tbo) in free-feeding unrestrained rats following injection into the medial hypothalamic paraventricular nucleus (PVN) or the lateral perifornical hypothalamus (PFH). NPY (78-235 pmol) or saline was infused unilaterally into the PVN or PFH in a volume of 0.4 microliter and simultaneous measures of food intake and Tbo were taken every 30 min for 3 h. Results indicated that NPY evoked changes in eating behavior and Tbo that were dependent upon the site of hypothalamic injection. Although PVN and PFH administration of NPY both increased food intake dose dependently within 30 min of treatment, PFH NPY-injected rats (n = 9) showed a stronger behavioral response compared to rats (n = 9) receiving NPY injections into the PVN. In PVN-treated rats, however, the increased eating was associated with a significant decline in Tbo evident within the first 30-min test interval. A mean maximal decline of 0.92 +/- 0.26 degree C occurred within 90 min of PVN treatment of the highest dose, which produced a reduction in Tbo that was maintained for 2.5 h. In contrast, NPY infusion into the PFH failed to reliably alter Tbo at any of the doses tested. These findings are consistent with evidence that NPY in the PVN and PFH may have distinct functions and suggest that although PFH NPY acts to stimulate a robust and relatively specific ingestive response, PVN NPY may participate in the complex integrative mechanisms responsible for the simultaneous regulation of feeding, thermoregulatory, and metabolic processes. PMID- 7479291 TI - Alpha-melanocyte-stimulating hormone (MSH) inhibits insulin secretion in HIT-T 15 cells. AB - alpha-Melanocyte-stimulating hormone (MSH) antagonizes the binding of interleukin 1 beta (IL-1) to its type I receptors on T cells. IL-1 modulates insulin secretion in the insulin-secreting tumor cell line, HIT-T 15 cells. In the present studies, we examined a hypothesis that alpha-MSH may modulate insulin secretion in HIT-T 15 cells. The addition of alpha-MSH over the concentrations of 10(-10) M inhibited insulin secretion for 4 h. In addition, the carboxyl-terminal tripeptide of alpha-MSH [alpha-MSH(11-13)], which is supposed to be an important sequence of alpha-MSH on its antipyretic effects, significantly inhibited insulin secretion at the concentration of 10(-9) M. In contrast, acetyl-[Nle4,D-Phe7] alpha-MSH(4-10)-amide (10(-11) to 10(-9) M) failed to inhibit insulin secretion. The maximal effect of alpha-MSH on insulin secretion was obtained at the concentration of 10(-9) M. The addition of alpha-MSH from 10(-11) to 10(-9) M significantly inhibited 7.5 mM glucose-stimulated insulin secretion at each period of 0-5, 5-15, and 15-30 min. Radioreceptor assay suggested that HIT-T 15 cells possess the specific binding sites for 125I-labeled alpha-MSH. The present studies indicated that alpha-MSH inhibits insulin secretion through the specific binding site in HIT-T 15 cells. PMID- 7479293 TI - Tachykinins with unusual structural features from a urodele, the amphiuma, an elasmobranch, the hammerhead shark, and an agnathan, the river lamprey. AB - Tachykinins were purified from extracts of gastrointestinal tissues of the urodele, Amphiuma tridactylum (three-toed amphiuma), and the elasmobranch Sphyrna lewini (hammerhead shark), and from the brain of the agnathan Lampetra fluviatilis (river lamprey). The amphiuma substance P (SP) (DNPSVGQFYGLM-NH2) contains 12 amino residues compared with 11 for mammalian SP and lacks the Arg/Lys-Pro-Xaa-Pro motif that is characteristic of NK1 receptor-selective agonists. Lampetra SP (RKPHPKEFVGLM-NH2) is identical to SP from the sea lamprey and the shark SP-related peptide (AKFDKFYGLM-NH2) is identical to dogfish scyliorhinin I. Amphiuma neurokinin A (NKA) (HKDAFIGLM-NH2) and lamprey NKA (HFDEFVGLM-NH2) contain 9 amino acid residues compared with 10 for mammalian NKA. The shark NKA-related peptide (ASGPTQAGIV10GRKRQKGEMF20VGLM-NH2) shows limited structural similarity to mammalian neuropeptide gamma and the teleost tachykinin, carassin but contains 24 rather than 21 amino acid residues. The data show that the primary structures of the tachykinins have been very poorly conserved during vertebrate evolution and that pressure has acted only to maintain the functionally important sequence -Phe-Xaa-Gly Leu-Met-NH2 at the COOH-termini of the peptides. PMID- 7479292 TI - Parallel bioassay of PG-SPI, an amphibian acidic SP-like peptide, mammalian basic substance P, and neurokinins A and B on in vitro and in vivo test systems. AB - In vitro and in vivo test systems were used to compare the biological activities of substance P and the neurokinins A and B with those of a newly isolated substance P-like acidic peptide, PG-SPI (pGlu-Pro-Asn-Pro-Asp-Glu-Phe-Phe-Gly-Leu Met-NH2). On nearly all the isolated smooth muscle preparations tested, PG-SPI appeared only slightly more potent than SP, but on guinea pig trachea it was 50 times more potent. The in vitro spasmogenic effect of PG-SPI on guinea pig trachea was inhibited by the NK1 receptor antagonist, CP 96,345. In in vivo tests, intracerebroventricularly injected PG-SPI was about 20 times more potent than SP in inhibiting gastric acid secretion and emptying in rats. Tests with antagonists showed that CP 96,345 reduced PG-SPI-induced inhibition of gastric emptying and the NK2 receptor antagonist, MEN 10,376, sharply blocked PG-SPI induced inhibition of gastric acid secretion. These findings fit poorly into the current classification of tachykinin receptors and suggest that the acidic substance P-like peptide is a valuable tool for studying the functional role of other tachykinin receptor subtypes. PMID- 7479294 TI - Dynorphin A(1-8) in human placenta: amino acid sequence determined by tandem mass spectrometry. AB - Presence of the kappa receptor-preferring neuropeptide dynorphin A(1-8) in human placenta has been demonstrated by mass spectrometry to establish rigorously the appropriate molecular weight and amino acid sequence. Liquid secondary ionization mass spectrometry produced the protonated molecule ion, (M + H)+, at m/z 981 of the endogenous peptide, and tandem mass spectrometry collected the product ion spectrum that contained the appropriate amino acid sequence-determining fragment ions produced from the precursor ion (M + H)+. The amino acid sequence of the peptide is YGGFLRRI. PMID- 7479295 TI - Inhibitory effect of zinc-chelating dipeptide on parathyroid hormone-stimulated osteoclast-like cell formation in mouse marrow cultures: involvement of calcium signaling. AB - A possible mechanism of zinc action inhibiting the PTH-induced osteoclast-like cell formation in mouse marrow culture system in vitro was investigated. Bone marrow cells were cultured for 7 days in alpha-minimal essential medium containing a well-known bone-resorbing agent parathyroid hormone (1-34) (PTH). Osteoclast-like cell formation was estimated with staining for tartrate-resistant acid phosphatase (TRACP), a marker enzyme of osteoclasts. The effect of zinc sulfate (10(-6) M) or beta-alanyl-L-histidinato zinc (AHZ; 10(-6) M) inhibiting the PTH (10(-8) M)-induced osteoclast-like cell formation was clearly seen in the absence or presence of theophylline (10(-4) M). However, zinc compounds did not inhibit the stimulatory effect of dibutyryladenosine 3',5'-cyclic monophosphate (DBcAMP; 10(-4) M) on osteoclast-like cell formation. The stimulating effect of PTH (10(-8) M) on osteoclast-like cell formation was clearly weakened (about 50%) in the presence of EGTA (1.0 mM) or dibucaine (10(-5) M). Phorbol 12-myristate 13 acetate (PMA), a protein kinase C activator (10(-8) to 10(-6) M), clearly stimulated osteoclast-like cell formation. PMA effect was inhibited by the presence of AHZ (10(-6) M) or zinc sulfate (10(-8) M). However, the inhibitory effect of zinc compounds was not seen in the presence of both PTH (10(-8) M) and PMA (10(-6) M). The present findings suggest that zinc compounds inhibit PTH stimulated osteoclast-like cell formation mediated through the Ca(2+)-dependent activation of protein kinase C. PMID- 7479296 TI - Effect of BCH 325 (Pro-D-Phe-Pro-Gly) on central dopaminergic functions. AB - Three behavioral models were used to characterize the pharmacological action of BCH 325 on central dopaminergic transmission. The effect of acute SC treatment with BCH 325 upon dopaminergic mechanisms affecting motor activity was studied on the climbing behavior of mice. It was shown that the beta-casomorphin analogue evoked a dose-dependent increase in apomorphine (APO)-induced hypoactivity that was reversed by sulpiride (SULP). In in vitro studies on slices of nucleus accumbens of mice it could be demonstrated that 10(-6) M APO caused a reduction of K(+)-stimulated [14C]dopamine (DA) release that was potentiated following simultaneous incubation with 10(-6) M BCH 325. To prove a postsynaptic influence in D1 receptor-mediated behavior pattern, the action of BCH 325 was studied on bromocriptine (BROMO)-evoked yawning behavior of rats after pretreatment with reserpine (RES) or saline. The peptide could not influence the BROMO yawning after saline administration, but it was able to normalize the number of yawns, which were reduced after RES. To investigate the effect of BCH 325 on postsynaptic D2 receptors, jerking behavior on RES-pretreated rats after a high dose of BROMO was used. Following RES pretreatment only, the number of BROMO induced jerks was decreased by treatment of rats with 0.5 mumol/kg BCH 325. In contrast, the jerking behavior was enhanced by 0.5 mumol/kg BCH 325 in rats that were additionally treated with alpha-methyl-p-tyrosine (MPT). In biochemical studies on slices of the nucleus accumbens of mice, the in vivo pretreatment with RES caused a reduction of K(+)-stimulated [14C]DA release that was blocked by the SC administration of 0.5 mumol/kg BCH 325.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7479297 TI - Prolonged effects of tumor necrosis factor-alpha on anterior pituitary hormone release. AB - We examined the chronic (72 h) effects of 30 ng/ml recombinant murine tumor necrosis factor (TNF)-alpha on release of immunoreactive growth hormone (GH), prolactin (PRL), thyrotropin (TSH), and TSH glycosylation, as assessed by lectin binding, in cultured rat anterior pituitary cells. In cultured cells from adult female rats, TNF-alpha significantly suppressed basal and GH-releasing hormone (GRH)-stimulated GH release. TNF-alpha also suppressed basal PRL release and completely abolished the PRL response to TRH (0.1-10 nM). Whereas TNF-alpha reduced basal TSH release, it significantly enhanced the maximal TSH response to TRH. TNF-alpha did not affect the concanavalin A and lentil lectin binding of TSH accumulated in the medium during the 4-day culture, but significantly decreased the lentil lectin binding of TSH released in response to acute TRH stimulation. TNF-alpha significantly enhanced the inhibitory effect of somatostatin on stimulated PRL release, but not on GH or TSH release. Compared to cell cultures from adult female rats, in anterior pituitary cell cultures from 12-day-old rats the effects of prolonged exposure to TNF-alpha on hormone release were diminished or absent. Pituitary hormone release was unaffected by acute (3 h) exposure to TNF-alpha. These results demonstrate a direct effect of TNF-alpha on anterior pituitary hormone release, which is cell-type specific and age dependent. PMID- 7479298 TI - Neutral endopeptidase can hydrolyze beta-amyloid(1-40) but shows no effect on beta-amyloid precursor protein metabolism. AB - High performance liquid chromatographic analyses of incubations of beta-amyloid(1 40) with neutral endopeptidase revealed at least nine product peaks, indicating that neutral endopeptidase can cleave beta-amyloid at multiple sites. Mass spectroscopic analysis of hydrolyzed beta-amyloid identified at least five cleavage sites, between residues Glu3-Phe4, Gly9-Trp10, Phe19-Phe20, Ala30-Ile31, and Gly33-Leu34. In contrast, amyloid precursor protein metabolism in Neuro2A cells was unaffected by the expression of recombinant neutral endopeptidase in the same cells or by the addition of a secreted form of neutral endopeptidase to spent Neuro2A cell media. PMID- 7479299 TI - Plasma atrial natriuretic peptide during spontaneous bronchoconstriction in asthmatics. AB - The aim of this work was to establish the role of endogeneous ANP during a spontaneous asthma attack. Forced expiratory lung volume in 1 s (FEV1), cardiovascular parameters, and plasma ANP, cAMP, and cGMP were measured for 60 min before and 10 min after treatment with a bronchodilator in 10 asthmatics. The results show that in the presence of moderate bronchoconstriction, FEV1 was 54 +/ 3% (+/-SE); ANP levels initially were slightly elevated at 47 +/- 10 pg/ml and decreased to 26 +/- 3 pg/ml (p < 0.05) over 60 min, with no change in FEV1. Following salbutamol inhalation, FEV1 increased to 77 +/- 4% with no change in ANP. We conclude that endogenous ANP does not act as a bronchodilator in asthmatics with moderate bronchospasm. PMID- 7479300 TI - Changes in late auditory evoked potentials induced by growth hormone-releasing hormone (GHRH) but not somatostatin (SRIF) after peripheral administration in male controls. AB - To investigate possible influences of growth hormone-releasing hormone (GHRH) and somatostatin (SRIF) on auditory perceptional processes, 12 subjects received either placebo (sodium chloride 0.9%), GHRH (50 micrograms), or SRIF (100 micrograms) on different days. Late auditory evoked potentials (AEP) were computed and further analyzed by using the brain electric source analysis (BESA) method. Reduced late AEP latencies were observed following GHRH administration. In contrast, SRIF had no significant effects on the AEP. The changes in late auditory processing seen after administration of GHRH were most likely induced by a direct central nervous action. PMID- 7479301 TI - Proopiomelanocorticotropin (POMC) peptides and lipoprotein lipase activity in vitro. AB - Effects of alpha-melanocyte-stimulating hormone (alpha-MSH), beta-melanocyte stimulating hormone (beta-MSH), beta-lipotropin (beta-LPH), and beta-endorphin (beta-EPH) at concentrations from 10(-9) M up to 10(-6) M on human adipose tissue lipoprotein lipase (LPL) were studied in a cell-free system. alpha-MSH and beta MSH did not exert any effect on LPL; no degradation of these peptides in the incubation medium could be detected by HPLC analysis. beta-LPH and beta-EPH failed to alter enzyme activity. However, HPLC analysis revealed an unspecific rapid degradation of the peptides due to the activity of tissue proteases. Therefore, the protease inhibitors amastatin, antipain, APMSF, and TPCK were tested at concentrations of 10(-5), 10(-4), and 10(-3) M for their efficacy to inhibit degradation. None of the inhibitors was able to substantially reduce proteolysis of beta-LPH, as was the case with amastatin, APMSF, and TPCK for beta EPH. However, antipain at 10(-4) M preserved at least 20% of the initial peptide concentration from proteolysis up to 150 min. Antipain caused a decrease in lipoprotein lipase activity (LPLA), which was dependent on concentration. The adverse effect of antipain at concentrations of 10(-4) M on LPL was completely abolished by beta-EPH at a concentration of 10(-6) M. PMID- 7479302 TI - Autoradiographic localization of NK1 and NK3 tachykinin receptors in rat kidney. AB - The distribution of neurokinin receptors in rat kidney, renal artery, renal vein, and proximal ureter was evaluated by autoradiography after in vitro labeling of NK1 sites with [125I]Bolton-Hunter substance P (BHSP) or NK3 sites with [125I][MePhe7]neurokinin B ([MePhe7]NKB). Film autoradiography using [125I][MePhe7]NKB revealed specific binding sites associated with the renal vein and its large branches, the renal pelvis, the inner strip of outer renal medulla, and the proximal ureter. High-resolution autoradiograms demonstrated that these sites were localized to the smooth muscle layer in the veins, pelvis, and ureter. Neither the renal arterial system nor the renal cortex contained specific [125I][MePhe7]NKB binding sites although a high level of nonspecific binding was associated with the renal artery. Specific binding of [125I]BHSP was associated with the renal artery and renal pelvis but not the renal veins. Arterial NK1 receptors appeared to be localized to the adventitia. The results indicate that at least two types of tachykinin receptor are present in the rat kidney. The distinct localization observed for most of the NK1 and NK3 receptors suggests that they have different functions. PMID- 7479303 TI - Autoradiographic localization of calcitonin gene-related peptide receptors in guinea pig respiratory tract: effect of capsaicin pretreatment. AB - Calcitonin gene-related peptide (CGRP) is a potent vasodilator peptide present in capsaicin-sensitive neurons innervating the respiratory tract. In this study, the autoradiographic distribution of [125I]CGRP binding sites was investigated in guinea pig airways. Extremely dense specific binding occurred over parenchymal tissue, with moderate specific binding over tracheal glands, the endothelium of pulmonary veins and arteries, and small blood vessels in the bronchial wall. The localization of binding sites for [125I]CGRP over blood vessels but not bronchial smooth muscle correlates well with the physiological actions of this peptide, although the function of the parenchymal sites is unknown. No significant difference in binding was seen in vehicle- or capsaicin-pretreated animals, suggesting that sites are not reliant on factors from capsaicin-sensitive neurons. PMID- 7479304 TI - Proteolytic degradation of hemoglobin in erythrocytes leads to biologically active peptides. AB - A number of hemoglobin-derived homogeneous peptides were isolated from erythrocyte lysate. The amino acid sequences of nine peptides were determined. Seven out of nine peptides were relatively long, 30-32 membered peptides covering the N- or C-terminal sequences of globin chains. The remaining two were the pentapeptide neo-kyotorphin and its tetrapeptide derivative. PMID- 7479305 TI - Age-related changes in mitogen-induced beta-endorphin release from human peripheral blood mononuclear cells. AB - beta-Endorphin is an opioid peptide synthesized in the pituitary, hypothalamus, and immunocytes, known to affect immune responses both when added in vitro and when its synthesis is increased in vivo (e.g., during stress). We show here that, similar to its concentrations in peripheral blood mononuclear cells, the release of the opioid peptide from these cells after stimulation with polyclonal mitogens such as PHA or Con-A is also age dependent. Moreover, the effect of both mitogens on Ca2+ homeostasis changes with age. Finally, the ionophore ionomycin and the Ca2+ ATPase blocker thapsigargin induce the same age related effect on beta endorphin release. For these reasons, we suggest that calcium homeostasis might be important for the differences observed in the release of the opioid from cells obtained from younger (< or = 30 years) or older (> or = 45 years) volunteers. PMID- 7479306 TI - Integration of vagal afferent responses to duodenal loads and exogenous CCK in rats. AB - The neurophysiological responses to 0.1 ml duodenal balloon inflation, 0.5 ml duodenal loads of normal saline, and 100 pmol close celiac arterial infusions of cholecystokinin (CCK) were obtained from 14 left cervical vagal afferent fibers in 14 rats. Duodenal, but not gastric, loads increased discharge rates in these slowly adapting fibers. CCK alone excited these fibers, and CCK pretreatment amplified subsequent duodenal load responses. Furthermore, duodenal loads generated greater responses when combined with CCK infusions. The small (< 3 mm) receptive fields of these fibers were localized to the ventral wall of the proximal duodenum, with C fiber conduction velocities (< 2 m/s). These results demonstrate for the first time rat duodenal load-sensitive vagal afferents. They can integrate signals arising from CCK and duodenal loads, and may mediate aspects of the role of CCK in the inhibition of gastric emptying and the control of food intake. PMID- 7479307 TI - Expression of pituitary adenylate cyclase activating polypeptide and receptors in human brain tumors. AB - The capacity of brain tumor samples to synthesize pituitary adenylate cyclase activating polypeptide (PACAP) was evaluated by the reverse transcriptase polymerase chain reaction technique (RT-PCR). The expression of PACAP receptors was assessed by a combination of RT-PCR techniques, conventional binding techniques, and also by the ability of PACAP to stimulate adenylate cyclase activity. A weak PACAP mRNA and PACAP receptor mRNA expression was detected in only 3 of 16 meningiomas. A weak PACAP-stimulated adenylate cyclase activity (+20%) was detected in 10 of the 16 samples but binding of labeled PACAP was never observed. In the 16 gliomas studied (including two oligodendrogliomas and two ependymomas), PACAP mRNA was identified in 13 samples and PACAP receptor mRNA in 15 samples. PACAP receptors were identified in all the samples by binding studies and/or by PACAP stimulation of the adenylate cyclase activity. PACAP mRNA was never detected in pituitary adenomas (three prolactinomas, two mixed PRL-GH producing tumors, three GH-secreting tumors, three gonadotrophinomas, one ACTH producing tumor, two nonsecreting tumors) whereas PACAP receptor mRNA was highly expressed in all the tumors except prolactinomas, where it was at the limit of detection, confirming the binding and adenylate cyclase activation results. Thus, it is unlikely that the neuropeptide PACAP could influence meningioma's cell growth; PACAP secreted from extratumoral areas may influence pituitary tumors and PACAP could participate to gliomas development. PMID- 7479308 TI - Substance P induces the expression of intercellular adhesion molecule-1 on vascular endothelial cells and enhances neutrophil transendothelial migration. AB - To determine whether substance P induces the expression of adhesion molecules ICAM-1 and VCAM-1 on vascular endothelial cells, we examined the effect of substance P, its carboxyl- and amino-terminal peptides, and neurokinin A on the expression of ICAM-1 and VCAM-1 on cultured human umbilical vein endothelial cells (HUVEC). Substance P and the carboxyl-terminal peptide SP(6-11), but not the amino-terminal peptide SP(1-9) or neurokinin A, increased ICAM-1 expression on HUVEC. In contrast, substance P did not significantly induce VCAM-1 expression on HUVEC. Furthermore, stimulation of HUVEC with substance P increased neutrophil transendothelial migration through an ICAM-1-dependent mechanism, as indicated by the inhibitory effect of anti-ICAM-1 antibody. These results indicate that substance P induces ICAM-1 expression on vascular endothelial cells and thereby enhances neutrophil transendothelial migration. PMID- 7479309 TI - Central neuropeptide Y enhances bile secretion through vagal and muscarinic but not nitric oxide pathways in rats. AB - Neuropeptide Y (NPY) acts in the central nervous system to regulate gastrointestinal functions in rats and dogs. The effects of intracisternal injection of NPY on bile secretion and biliary components were investigated in urethane-anesthetized rats with bile duct cannula. Intracisternal NPY (0.02-0.12 nmol) dose-dependently increased bile secretion by 9.2-19.5%. The secretory response occurred within the first 20-40 min and lasted for the 120-min observation period. Intravenous injection of NPY (0.12 nmol) did not modify bile secretion under identical conditions. Biliary bile acid, phospholipid, and cholesterol secretion were not modified by intracisternal injection of NPY (0.12 nmol), whereas bicarbonate was increased by 19.0 +/- 1.7% from 40 to 120 min after NPY injection. Cervical cord transection at the C6 level, acute bilateral adrenalectomy (-120 min), or injection of NG-nitro-L-arginine methyl ester (10 mg/kg, IV, -15 min), an inhibitor of nitric oxide biosynthesis, did not alter intracisternal NPY (0.12 nmol)-induced stimulation of bile secretion. Atropine (2.0 mg/kg, IP, -30 min) and bilateral cervical vagotomy (-120 min) completely abolished the stimulatory effect of intracisternal NPY (0.12 nmol) on bile secretion. These findings indicate that NPY acts in the brain to stimulate bicarbonate-dependent bile secretion through vagal and muscarinic pathways and suggest that peptides in the central nervous system may be involved in the vagal regulation of bile secretion. PMID- 7479310 TI - Characterization of receptors using cyanine 3-labeled neuropeptides. AB - We labeled substance P (SP), neurokinin A (NKA), and [Lys0]gastrin-releasing peptide-27 (GRP) with cyanine 3.18 (cy3). Cy3-peptides purified by HPLC were fully active, determined by [Ca2+]i mobilization. Binding was specific because it was abolished by unlabeled peptides and receptor antagonists. Transfected cells yielded a log-fold greater cy3 intensity than control cells by FACS. Confocal microscopy of transfected cells and cultured enteric neurons showed that cy3-SP bound to surface receptors and was internalized. Internalization was observed in living cells by capture of sequential images. Recovery of surface binding sites was monitored by flow cytometry using cy3-SP. Thus, cy3 neuropeptides can be used to isolate and study receptor-bearing cells. PMID- 7479311 TI - Characterization of angiotensin II receptor subtypes in pancreatic acinar AR42J cells. AB - The AR42J acinar cell line was characterized as a potential cellular model to assess the functional aspects of an exocrine pancreatic angiotensin system. Binding studies revealed that the AR42J cells express high affinity angiotensin II binding sites (Kd = 0.73 +/- 0.06 nM; Bmax = 292 +/- 15 fmol/mg protein, n = 3). Competition studies established that these cells, similar to the intact pancreas, express predominantly the AT2 receptor subtype. The AT2-selective antagonists CGP 42112A, PD 123177, and PD 123319 competed for the majority of angiotensin II binding. However, 10-15% of the angiotensin II binding sites were competed for by the AT1-selective antagonist DuP 753 (Losartan). Affinity labeling of these binding sites with [125I]angiotensin II followed by SDS gel electrophoresis under reducing conditions revealed a single band comprising a molecular mass of 108,000 Da. Competition with unlabeled angiotensin II or the AT2 antagonist, but not the AT1 antagonist, abolished the 108,000-Da band. In intact cells, angiotensin II caused a rapid increase in intracellular calcium (Ca2+) using Fura-2 as a Ca2+ indicator. Pretreatment of the cells with the AT1 antagonist DuP 753 completely inhibited the angiotensin II-induced rise in Ca2+; however, the AT2 antagonists CGP 42112A and PD 123177 were ineffective in blocking the Ca2+ increase. These results demonstrate that this pancreatic acinar cell line expresses both AT2 and AT1 angiotensin II receptor subtypes. The AT1 receptor is coupled to the mobilization of Ca(2+)--a characteristic shared by AT1 receptors in other tissues. PMID- 7479312 TI - Immunoprecipitation and characterization of a binding protein specific for the peptide, intestinal trefoil factor. AB - Recombinant rat intestinal trefoil factor (rITF) and human spasmolytic polypeptide (hSP) were irreversibly cross-linked to specific binding sites in solubilized rat intestinal epithelial membranes and human adenocarcinoma cells. Analysis of the immunoprecipitates by immunoblotting identified a cross-linked protein complex of approximately 45 kDa, which under reducing conditions appeared as a approximately 28-kDa band and the latter displayed ligand-stimulated phosphorylation of a tyrosine, but not a threonine or serine, residue in the binding complex. [125I]rITF was used to localize binding sites by autoradiography of frozen sections from rat gastrointestinal tissues. A high density of specific [125I]rITF binding sites was present within gastric, colonic, and jejunal mucosal glands. Unlabeled hSP partially inhibited [125I]rITF binding at a concentration of 1 microM when compared with the same concentration of unlabeled rITF. These studies support earlier observations for the existence of trefoil binding sites in the gastrointestinal tract and further suggest that hSP has affinity for the mucosal rITF binding site. PMID- 7479313 TI - Neuropeptide Y, the hypothalamus, and diabetes: insights into the central control of metabolism. AB - Neuropeptide Y (NPY), a major brain neurotransmitter, is expressed in neurons of the hypothalamic arcuate nucleus (ARC) that project mainly to the paraventricular nucleus (PVN), an important site of NPY release. NPY synthesis in the ARC is thought to be regulated by several factors, notably insulin, which may exert an inhibitory action. The effects of NPY injected into the PVN and other sites include hyperphagia, reduced energy expenditure and enhanced weight gain, insulin secretion, and stimulation of corticotropin and corticosterone release. The ARC PVN projection appears to be overactive in insulin-deficient diabetic rats, and could contribute to the compensatory hyperphagia and reduced energy expenditure, and pituitary dysfunction found in these animals; overactivity of these NPY neurons may be due to reduction of insulin's normal inhibitory effect. The ARC PVN projection is also stimulated in rat models of obesity +/- non-insulin diabetes, possibly because the hypothalamus is resistant to inhibition by insulin; in these animals, enhanced activity of ARC NPY neurons could cause hyperphagia, reduced energy expenditure, and obesity, and perhaps contribute to hyperinsulinemia and altered pituitary secretion. Overall, these findings suggest that NPY released in the hypothalamuss, especially from the ARC-PVN projection, plays a key role in the hypothalamic regulation of energy balance and metabolism. NPY is also found in the human hypothalamus. Its roles (if any) in human homeostasis and glucoregulation remain enigmatic, but the animal studies have identified it as a potential target for new drugs to treat obesity and perhaps NIDDM. PMID- 7479314 TI - Immunohistochemical localization of adrenomedullin in canine heart and aorta. AB - Adrenomedullin (ADM) is a new endogenous hypotensive and vasorelaxing peptide that may play an important role in the regulation of cardiovascular function. Although ADM was originally isolated from pheochromocytoma, ADM-like immunoreactivity has also been widely detected in various tissues, including the cardiovascular system. Based upon the reports that ADM mRNA and ADM-like immunoreactivity are present in the heart, the present study was designed to investigate the immunohistochemical localization of ADM in the canine heart and aorta. In the canine heart, immunohistochemical examination revealed positive immunostaining within the myocardia in both atria and ventricles. ADM immunoreactivity was observed within the cytoplasm of myocardium, and was widely distributed in the peripheral cytoplasm. ADM immunoreactivity was more intense in the atria than in the ventricles. In the canine aorta, vascular smooth muscle cells of the aorta and vasa vasorum were also immunopositive for ADM. ADM immunoreactivity was mostly localized in the perinuclear position within the smooth muscle cells. There was no immunoreactivity in endothelium, endocardium, epicardium, adventitia, or connective tissues. The current study demonstrates for the first time that immunoreactive ADM by immunohistochemistry is present in the cardiovascular system. As ADM has hypotensive and vasorelaxing actions and circulates in the body, ADM is a cardiovascular peptide hormone that may play an important role in the regulation of cardiovascular system. PMID- 7479315 TI - Metabolism of the anticancer peptide H-Arg-D-Trp-NmePhe-D-Trp-Leu-Met-NH2. AB - H-Arg-D-Trp-NmePhe-D-Trp-Leu-Met-NH2 (Antagonist G) will be the first broad spectrum neuropeptide antagonist to enter a phase I clinical trial. Its in vitro and in vivo metabolism has been extensively characterized. The major metabolites were identified and their structures elucidated by mass spectroscopy and amino acid analysis. Metabolism occurred almost exclusively at the C-terminus and was arrested by phenylmethylsulfonylfluoride, a known serine-protease inhibitor. Biological characterization of the metabolites demonstrated that the degradation of Antagonist G produces metabolites that retain neuropeptide antagonist properties. PMID- 7479317 TI - Distribution, formation, and molecular forms of the peptide xenin in various mammals. AB - In the present investigation we isolated the recently discovered pentacosapeptide xenin from gastric mucosa of man, dog, pig, guinea pig, rat, and rabbit. HPLC, mass spectrometry, and amino acid sequence analysis showed xenin-25 in concentrations of 54-144 pmol/g tissue in gastric mucosa of each species. Extraction with 2% TFA followed by analytical C18 HPLC revealed 0.02-84 pmol/g xenin-25 also in hypothalamus, lung, liver, heart, kidney, adrenal gland, pancreas, testicle, skin, and duodenal, jejunal, ileal, and colonic mucosa of dog and man, respectively. Digestion of these acid extracts with pepsin liberated xenin-25 in concentrations from 2 up to 166 pmol/g tissue. Gel chromatography revealed a large molecular weight precursor of xenin-25 and evidence for an endogenous acid protease coeluting with pepsinogen capable of releasing xenin-25 from its precursor. Maximal concentrations of xenin-25 were obtained when canine gastric mucosa was incubated with 2% TFA at room temperature for 2 h. Longer incubation times led to a decline of xenin-25 concentration and to formation of xenin-16 and xenin-9, both C-terminal fragments of xenin-25. We conclude that xenin-25 is present not only in human gastric mucosa but also in the stomach of various other mammals. Xenin-25 is further present in low concentrations in many other organs where a pepsin-like protease generates xenin-25 from a large precursor and processes it to smaller fragments. PMID- 7479316 TI - Isolation and characterization of ovokinin, a bradykinin B1 agonist peptide derived from ovalbumin. AB - A vasorelaxing peptide was purified from a peptic digest of ovalbumin, after three steps of reverse-phase HPLC. The structure of the peptide was Phe-Arg-Ala Asp-His-Pro-Phe-Leu, which corresponded to residues 358-365 of ovalbumin. The peptide was named ovokinin. Ovokinin showed relaxing activity for a canine mesenteric artery (EC50 = 6.3 microM). The relaxing activity was blocked by the bradykinin B1 antagonist [des-Arg9] [Leu8]bradykinin, but not by the B2 antagonist Hoe 140. Ovokinin binds to B1 receptors (IC50 = 64 microM). Prostaglandin I2 was released from the artery after ovokinin stimulation as a relaxing factor. Thus, ovokinin is a weak bradykinin B1 agonist peptide derived from food proteins. PMID- 7479318 TI - Analysis of the peptide content of the locust vasopressin-like immunoreactive (VPLI) neurons. AB - Isolated cell bodies of the locust vasopressin-like immunoreactive (VPLI) neurons, analyzed by HPLC separation and radioimmune assay, contain three arginine vasopressin-like peptides: a previously identified monomer (Fl, Cys-Leu Ile-Thr-Asn-Cys-Pro-Arg-Gly-NH2) and its antiparallel homodimer (F2), but also the previously unreported parallel homodimer (PDm). VPLI neuron activity significantly reduces the level of cAMP in the CNS. Of the three synthetic peptides, only the monomer (F1, 10(-8) and 10(-6) M) is capable of inhibiting a forskolin-stimulated increase in cAMP in isolated neural membranes. The antiparallel (F2) and parallel dimers (PDm) of this peptide have no effect on this second messenger. PMID- 7479319 TI - Diuretic activity of C-terminal group analogues of the insect kinins in Acheta domesticus. AB - A series of insect kinin analogues, AFFPWG-X, modified at the C-terminal group, were evaluated in a cricket Malpighian tubule secretion bioassay. The results were compared with activity profiles observed in a cockroach hindgut myotropic bioassay for these analogues. Although the replacement of the C-terminal amide group with a negatively charged acid leads to a precipitious drop in diuretic activity, it can be partially restored with the introduction of ester groups such as methyl or benzyl. The presence of branched chain character in the C-terminal group or a C-terminal alpha-carbon-amide distance spanning five methylene group spacers is incompatible with the receptor interaction required for biological activity. Significant diuretic activity is retained with four or fewer methylene groups in this region. C-terminal group analogues containing -SCH3, NHCH2CH2OCH3, or -OCH2(C6H5) offered the greatest retention of diuretic activity while providing increased hydrophobicity and/or steric bulk. The data are of potential value in the development of mimetic analogues of this insect neuropeptide family. Mimetic analogues are potentially valuable tools to insect neuroendocrinologists studying diuresis and/or engaged in the development of future pest management strategies. PMID- 7479320 TI - Behavioral effects of the cyclic cholecystokinin peptide analogue JMV-320. AB - The in vivo effects of JMV-320 (a highly selective CCKB receptor ligand) and of CCK-4 on exploratory activity and memory in rats were compared. JMV-320 and CCK-4 did not modulate exploratory activity in an open field test but decreased it in an elevated plus-maze. CCK-4 (50 micrograms/kg) impaired passive avoidance response 3 h after training and JMV-320 (1 and 10 micrograms/kg) decreased active avoidance response 24 h after training. The behavioral effects of JMV-320 resemble the effects of CCK-4 and suggest that in vivo JMV-320 acts as a CCKB receptor agonist. PMID- 7479321 TI - alpha-MSH-induced behavior: changes after diazepam and baclofen administration related with cyclic AMP levels. AB - The present work was performed to evaluate the participation of the benzodiacepinic GABAA and GABAB components upon excessive grooming, locomotion, rearing, and stretching/yawning syndrome induced by the intracerebroventricularly alpha-MSH administration by using GABAA and GABAB agonists. It also aims at evaluating possible relation between changes in cAMP levels in caudate-putamen and accumbens nuclei and the behavioral responses. Injection of diazepam or baclofen reduced the total behavioral scores in a dose-related manner as well as the cAMP levels with respect to the control values (animals treated with artificial cerebrospinal fluid). When diazepam was tested in animals simultaneously injected with alpha-MSH, behavioral scores decreased with respect to those treated with the peptide alone. Cyclic AMP also decreased after combined treatment (MSH + diazepam). PMID- 7479322 TI - Developmental changes of tachykinins in the hypothalamus and anterior pituitary of female Siberian hamsters from prepuberty to adulthood. AB - The developmental changes of hypothalamic and anterior pituitary tachykinin concentrations were studied in female Siberian hamsters kept either under short (SD) or long (LD) daily photoperiods. The animals were killed between 15 and 70 days of age. Hypothalamic NKA sharply increased starting at 15 days up to 50 days in LD animals, and between 20 and 60 days in SD animals. Hypothalamic SP levels increased in a similar manner in SD animals, but in LD animals the increment was less pronounced, with increased levels from day 20 to 40, followed by a plateau. In the anterior pituitary gland, NKA concentrations in LD animals increased at 40 days of age, with only slight increases afterward, but overall the increment curve was considerably flatter than for hypothalamic NKA. In SD animals, the increase of anterior pituitary NKA was much steeper than in LD animals. However, the total content of NKA in the AP was similar in both SD and LD animals, because the AP weight was considerably higher in LD- than in SD-exposed hamsters. These results showed that photoperiod did not markedly affect the developmental changes in hypothalamic NKA. The developmental changes in anterior pituitary NKA concentrations were considerably smaller than in the hypothalamus in LD animals, but in SD animals they were much steeper. NKA concentrations in the anterior pituitary were markedly affected by the photoperiod. Concentrations of NKA in the anterior pituitary of the Siberian hamster at the age of 15 days of age were already higher than in the anterior pituitary of adult rats or Syrian hamsters.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7479323 TI - Fluorescence polarization assay for endothelin-converting enzymes. AB - The conversion of big endothelin to endothelin by alpha-chymotrypsin was determined by following its single Trp fluorescence polarization. This provides a novel, simple, fast, and sensitive identifying assay in the search for a native endothelin-converting enzyme. PMID- 7479325 TI - Comparison of hCRF and oCRF effects on cardiovascular responses after central, peripheral, and in vitro application. AB - Three assays have been used to show that the neuropeptides human corticotropin releasing factor (hCRF) and the ovine analogue oCRF produced substantial dose dependent cardiovascular responses. The assays included intracerebroventricular (ICV) and intravenous (IV) administration in conscious rats, and also in vitro experiments with resistance arteries. Central administration of the peptides (0.1 10 micrograms, ICV) caused an increase in blood pressure and heart rate, whereas peripheral administration (0.75-750 micrograms/kg, IV) produced a decrease in blood pressure and tachycardia. Isometric ring preparations of mesenteric resistance arteries (diameter 200 microns) relaxed in response to both peptides (1-100 nM). In all cases, the effects were more pronounced for hCRF compared to compared to oCRF. Furthermore, all effects were inhibited by the CRF analogue alpha-helical CRF(9-41), the effect of the analogue being most potent against oCRF. The results of all three assays indicate that the difference in structure between hCRF and oCRF produces differences in biological activity. PMID- 7479326 TI - A new tritiated oxytocin receptor radioligand--synthesis and application for localization of central oxytocin receptors. AB - A new tritiated oxytocin antagonist radioligand was synthesized by introducing a tritiated propionic acid residue into the free amino group of ornithine in position 8 of the parent peptide [1-(beta-mercapto-beta,beta-cyclopentamethylene propionic acid), 2-(O-methyl)-tyrosine, 4-threonine, 8-ornithine, 9 tyrosylamide]vasotocin (OTA), that was previously described. The tritiated compound [3H][1-(beta-mercapto-beta,beta-cyclo-pentamethylene propionic acid), 2 (O-methyl)-tyrosine, 4-threonine, 8-(N6-propionyl)-ornithine, 9 tyrosylamide]vasotocin ([3H]PrOTA) was obtained in good yield with high specific activity (100 Ci/mmol). [3H]PrOTA exhibited the same affinity (Kd = 0.8 nM) and selectivity for the myometrial oxytocin receptor as the iodinated antagonist [125I]OTA. Autoradiographic localization of oxytocin receptors in the rat brain showed specific binding sites for [3H]PrOTA within regions of the limbic system, the neocortex, and hypothalamus, which is consistent with the binding pattern obtained with [125I]OTA. The high specific activity in combination with the long half-life of tritium and its low radiotoxicity as compared to iodine-125 makes the new tritiated antagonist a valuable tool for pharmacological studies. PMID- 7479324 TI - Rat corticostatin R4: synthesis, disulfide bridge assignment, and in vivo activity. AB - We have synthesized significant amounts of the most potent member of the rat corticostatins that inhibits ACTH-induced corticosteroid and compared its structure to that of the natural hormone. The cystine bridging arrangement that corresponds to that reported for a human defensin (3-31, 5-20, 10-30) was determined. The in vitro corticostatic activity of the synthetic rat corticostatin R4 paralleled that of the natural R4. Biological studies in vivo showed that doses of 8 or 12 mg corticostatin/kg effectively interfered with corticosterone release in stressed rats. We conclude that in the assays that were used, the biological activity of the synthetic and natural molecules was identical. The availability of significant amounts of synthetic material will make possible studies investigating the physiological role played by corticostatins in modulating the activity of the hypothalamic-pituitary-adrenal axis. PMID- 7479327 TI - ELISA measurements of vasotocin and isotocin in plasma and pituitary of the rainbow trout: effect of salinity. AB - The fish neurohypophysial hormones arginine vasotocin (AVT) and isotocin (IT) were measured for the first time by ELISAs (in comparison with other techniques) in plasma and hypophysis of rainbow trout adapted stepwise from freshwater (FW) to seawater (SW). AVT concentrations were higher than IT in plasma and, conversely, lower in hypophysis. No difference appeared between FW and SW conditions, but plasma hormone concentrations fell when FW fish were moved to 1/3 SW and increased progressively when fish were moved from 1/3 SW to SW. Peptide values obtained in 1/3 SW may correspond to the lowest osmoregulatory constraints occurring in an isosmotic medium in comparison to FW or full SW. The data suggest that storage and/or release of AVT and IT differ, but vary in a similar way with external salinity, and that these peptides should play a role in teleost fish osmoregulation. PMID- 7479328 TI - Characterization and autoradiographic mapping of [3H]CP96,345, a nonpeptide selective NK1 receptor antagonist in guinea pig lung. AB - We have studied binding and distribution of NK1 receptors in guinea pig lung using [3H]CP96,345. Kinetic studies showed that specific binding of [3H]CP96,345 was rapid and reversible, giving a kinetic dissociation constant (Kd) of 0.28 +/- 0.05 nM. The specific binding was also saturable and Scatchard analysis indicated a single class of binding site with an equilibrium Kd of 0.12 +/- 0.03 nM and maximum binding capacity (Bmax) of 107.0 +/- 10.3 fmol/mg of protein. Competition studies showed the rank order of affinity for agonists and antagonists as follows: SP > NKA = septide >> NKB = senktide; CP96,345 > FK888 > FK224 > L668169. NK3 agonists, NK2-selective antagonists, and a calcium channel blocker, diltiazem, showed no displacement, indicating high selectivity for NK1 receptors. Autoradiographic mapping showed specific labeling over airway smooth muscle from central to peripheral airways, submucosal glands, and nerve fibers of trachea. The labeling of airway epithelium was increased with diminishing size of airways. Pulmonary blood vessels were also moderately labeled and there was sparse labeling over alveolar walls. [3H]CP96,345 may provide a useful tool to evaluate NK1 receptor expression in peripheral organs. PMID- 7479329 TI - Heterogeneity of neurokinin1 binding sites in porcine respiratory tract. AB - High-affinity NK1 binding sites for [125I]BH-SP were characterized in adult porcine respiratory tract. The affinity and density of NK1 sites were significantly higher in tracheal epithelium and smooth muscle than in the lung. The potency order for agonists was: SP > neuropeptide-gamma > physalaemin > NKA > eledoisin > septide > SP methyl ester > GR 73632 > NKB > senktide > SP(1-7). For antagonists: CP 99,994 > CP 96,345 > spantide > L 703,606 >> WIN 51,708. The CP compounds discriminated between very high- and high-affinity NK1 sites in all three tissues. The subpopulation of sites with very high affinity for CP compounds also preferentially bound septide. Both binding components were inhibited by guanine nucleotide and showed equal affinities for SP. We propose an NK1B subtype with very high affinity for the antagonists CP 99,994 and CP 96,345 and the agonist septide, and an NK1A subtype with lower affinities for these ligands. PMID- 7479330 TI - Immunocytochemical localization of motilin-containing cells in the rabbit gastrointestinal tract. AB - Motilin-immunopositive cells (Mo cells) are known to be present in the upper small intestine of various species, including man. However, whether Mo cells are present in the rabbit gastrointestinal tract remained to be elucidated. Therefore, this study was designed to investigate the distribution of Mo cells in the rabbit gastrointestinal tract by the avidin-biotin-peroxidase complex method using a new anti-motilin serum (CPV2) raised in chickens. The results of an enzyme-linked immunosorbent assay suggested that this antiserum recognized the C terminal region of the motilin molecule. Motilin-immunopositive cells were found in the epithelia of the crypts and villi throughout the rabbit gastrointestinal tract from the gastric antrum to the distal colon, but no immunostaining occurred in the gastric body. Morphometric analysis revealed that Mo cells were localized preferentially in the upper small intestine, as reported for other species, and the cell densities (cells/mm2, mean +/- SE) were: gastric antrum (0.41 +/- 0.16), duodenum (8.2 +/- 0.8), jejunum (1.9 +/- 0.5), ileum (0.62 +/- 0.14), cecum (0.19 +/- 0.05), proximal colon (0.13 +/- 0.03), and distal colon (0.39 +/- 0.18). Our results demonstrated conclusively that Mo cells exist in the rabbit gastrointestinal tract and showed for the first time their regional distribution. Furthermore, our new chicken antiserum would appear to be a useful tool for the determination of plasma motilin concentrations by radioimmunoassay and for the immunoneutralization of endogenous motilin in the rabbit. PMID- 7479331 TI - Neuropeptide Y inhibits Ca2+ oscillations, cyclic AMP, and secretion in melanotrope cells of Xenopus laevis via a Y1 receptor. AB - The melanotrope cells in the pituitary gland of Xenopus laevis are innervated by neurons containing neuropeptide Y (NPY). In the present study, the mechanism of action of NPY on the melanotropes has been investigated. NPY inhibited in vitro secretion from melanotropes in intact neurointermediate lobes as well as from isolated, single melanotropes. Inhibition of secretion from neurointermediate lobes was mimicked by the NPY analogues PYY and [Leu31,Pro34]NPY, whereas NPY(13 36) was inactive. Secretion from isolated melanotropes was inhibited by [Leu31,Pro34]NPY and NPY(13-36), but NPY(13-36) was 10-fold less potent than [Leu31,Pro34]NPY. Studies on isolated cells revealed that NPY and its analogues inhibited the occurrence of intracellular Ca2+ oscillations with the same potency as they inhibited secretion from isolated cells. In addition to inhibiting basal secretion and spontaneous Ca2+ oscillations, NPY inhibited the basal production of cyclic AMP. On the basis of these results it is proposed that NPY inhibits secretion from Xenopus melanotropes by inhibiting cyclic AMP-dependent spontaneous Ca2+ oscillations through a Y1-like receptor. PMID- 7479332 TI - [125I]endothelin binding in rat cerebellum is increased following L-2 chloropropionic-acid-induced granule cell necrosis. AB - The systemic administration of L-2-chloropropionic acid (L-CPA) to rats produced a marked depletion of cerebellar granule cells (> 80% of the total) when administered in a single oral dose of 750 or 250 mg/kg/day for 3 days. The nature of the L-CPA-induced neurotoxicity is currently unknown but it exhibits a number of features in common with excitatory amino acid-induced neuronal cell death. We observed an increase in [125I]endothelin-1 (ET-1) binding in the cerebellar cortex, as measured by quantitative receptor autoradiography, which occurs at 48 h, but not 24 h, following the 750 mg/kg L-CPA dosing regimen. The increase in [125I]ET-1 binding did not parallel the cellular damage and resultant astrocyte proliferation, as measured by GFAP immunoreactivity, which was primarily confined to the granular layer of the cerebellar cortex. The increased [125I]ET-1 binding occurred in the molecular layer of the cerebellar cortex (controls 2.03 +/- 0.26 fmol/mg tissue; L-CPA-treated 6.69 +/- 0.45 fmol/mg tissue, n = 6; p < 0.01, Student's t-test) which appeared to contain astrocytic processes originating from the large increase in astrocyte number situated in the granular layer. Pretreatment of the rats with the irreversible NMDA receptor antagonist, MK-801, protected the cerebellar granule cells against the L-CPA neurotoxicity and also prevented the increase in [125I]ET-1 binding in the cerebellar cortex. The increased [125I]ET-1 binding in rat cerebellum appears to be linked to the reactive gliosis that occurs in association with neuronal cell injury. PMID- 7479333 TI - Microstructural analysis of licking behavior following peripheral administration of bombesin or gastrin-releasing peptide. AB - To determine the effects of bombesin (BN) and gastrin-releasing peptide [GRP(1 27)] on food intake and the microstructure of licking, the rate and pattern of licking were measured by lickometer after intraperitoneal administration of 2, 4, 8, and 16 micrograms/kg of each peptide. Although both peptides inhibited intake significantly, the dose-response functions of the peptides were different. At lower doses (2-4 micrograms/kg) the peptides were equipotent; at higher doses (8 16 micrograms/kg) BN was significantly more potent than GRP(1-27), possibly due to an action of BN on neuromedin B (NMB) receptors. The microstructural results demonstrated that both peptides decreased intake by increasing the rate of decay of licking and decreasing the number of bursts and clusters without changing the initial rate of licking or the size of the bursts and clusters. This combination of microstructural changes is evidence that both peptides increased the postingestive negative feedback effects of the diet without changing the orosensory positive feedback. These results are consistent with, but do not prove, the hypothesis that endogenous GRP(1-27) acts as a postingestive satiety signal in rats. PMID- 7479334 TI - The effect of vasoactive intestinal peptide (VIP) on superoxide dismutase and catalase activities in renal tissues of rats exposed to hemorrhagic ischemia reperfusion. AB - The effect of vasoactive intestinal peptide (VIP) on the activities of superoxide dismutase and catalase was investigated in renal tissues of rats exposed to 30% hemorrhage followed by reperfusion. In addition to enzyme activities, renal tissues were also histologically evaluated. Thirty percent hemorrhage had no significant effect on the activity of either enzyme. Reperfusion altered the activity of renal catalase but not of superoxide dismutase. On the other hand, administration of VIP (25 ng.kg-1) together with shed blood retransfusion protected the renal tissue from hemorrhagic ischemia-reperfusion injury without increasing superoxide dismutase and catalase activity. These results seem to be related either to the inhibitory effect of VIP on production or quenching activity of some reactive oxygen species. In conclusion, VIP may be a novel promising therapeutic approach toward defenses against hemorrhagic ischemia reperfusion injury as an antioxidant. PMID- 7479335 TI - Interaction between long-term potentiation and the action of a peptide transmitter. AB - The interaction between long-term potentiation (LTP) and the action of a peptide transmitter was examined with evoked field potential recording in the CA1 region of the hippocampus in anesthetized male rats. The peptide transmitter transiently reversed and opposed the effects of LTP, and LTP reversed the action of the peptide transmitter. Because the peptide transmitter is released and has action particularly during sexual behaviors, the results are interpreted to mean that any memory trace encoded by potentiating mechanisms in the rat hippocampus is probably not accessible in the 20-30-min period of time surrounding reproductive activity in male rats. PMID- 7479336 TI - Immunolocalization of neuropeptide Y in human pancreatic endocrine tumors. AB - Neuropeptide Y (NPY) is a 36 amino acid peptide known to inhibit glucose stimulated insulin secretion. NPY has recently been shown to be synthetized within rat islets of Langerhans and to be secreted in a differentiated rat insulin-secreting cell line, and as to this date the localization of NPY in human endocrine pancreas has not been reported. As NPY shares high amino acid sequence homology with peptide YY (PYY) and pancreatic polypeptide (PP), the polyclonal antibodies raised against these peptides often cross-react with each other. To demonstrate the presence of NPY in the human endocrine pancreas, we used a highly specific monoclonal antibody raised against NPY and another against its C flanking peptide (CPON). We studied three cases of hyperplasia of Langerhans islets and 11 cases of endocrine tumors of the pancreas. NPY and CPON were detected in all three cases of hyperplasia. For the 11 pancreatic tumors, five and nine of the tumors were positive for the antibodies NPY and CPON, respectively. The two negative tumors for CPON immunoreactivity were differentiated insulinomas, which showed no evidence of other hormonal secretion. In normal Langerhans islet, NPY and CPON immunoreactivities were colocalized in glucagon-producing cells (alpha-cells) and in a few insulin-secreting cell (beta cells).(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7479337 TI - Pituitary adenylate cyclase-activating peptide stimulates neurite growth in PC12 cells. AB - The ability of PACAP-38 to stimulate morphological development was studied using rat pheochromocytoma PC12 cells. PACAP-38 produced concentration-dependent increases in percentage of cells exhibiting neurite extension. Similar increases were produced by forskolin (28 +/- 2% at 96 h) and 8-bromo cAMP (30 +/- 2%). Vasoactive intestinal peptide and alpha-calcitonin gene-related peptide were without effect. PACAP-38 produced significant increases in PC12 cell cAMP content and inositol phosphate turnover. Intracellular [Ca2+] increased from 169 +/- 14 nM to 560 +/- 58 nM in response to 1 microM PACAP-38. PACAP-stimulated neurite outgrowth was abolished by RpcAMPS, an inhibitor of cAMP-dependent kinases but was unaffected by the protein kinase C antagonist H7. PMID- 7479338 TI - Production of bioactive enkephalin from the nonendocrine cell lines COS-7, NIH3T3, Ltk-, and C2C12. AB - Enkephalin is synthesized from proenkephalin in neuroendocrine cells. For the attempt to induce nonneuroendocrine origin cells to produce enkephalin, we used a mammalian expression vector for fusion peptides, pMEproCT beta, in which a fused peptide is designed to be cleaved by a yeast Kex2-like endoprotease furin. Met Enkephalin was expressed in four nonneuroendocrine cell lines: COS-7, C2C12, Ltk , and NIH3T3. The four cell lines produced a marked amount of Met-enkephalin, which appeared as a single peak on reverse-phase HPLC. Because transplantation of adrenal medullary cells to the subarachnoid space has been used to alleviate terminal cancer pain, and enkephalin appears to play a central role in relieving pain, this enkephalin expression vector may be useful for direct enkephalin expression in pericancerous tissues. PMID- 7479341 TI - Seizure-induced activation of enkephalin- and somatostatin-synthesizing neurons. AB - The extent of the neuronal network that is activated by kainic acid-induced seizures was anatomically identified and neurochemically characterized. Seizure activated neurons were identified through the immunocytochemical demonstration of Fos protein in neuronal nuclei. These seizure-activated neurons were characterized by determining if they contained the mRNA for somatostatin or enkephalin, using in situ hybridization procedures. The results demonstrate that a majority of enkephalin- and somatostatin-synthesizing neurons expressed the Fos protein following seizures and that they represent a major component of the kainic acid-induced, seizure-activated neuronal network. PMID- 7479339 TI - Effect of carbachol on the release of peptide YY from isolated vascularly and luminally perfused rat ileum. AB - Possible cholinergic control on the release of PYY from intestine into the lumen or blood vessel was studied by radioimmunoassay in the isolated perfused rat ileum. The basal release of PYY into the lumen was 43.1 +/- 8.9 pg/min, which was comparable with that into the vasculature (35.2 +/- 2.6 pg/min). The administration of 1 microM carbachol into the vascular perfusate resulted in a more than 40-fold increase of the luminal release but only a twofold increase of the vascular release. Carbachol-induced release of PYY into both lumen and vasculature was completely blocked by atropine, but not by hexamethonium. Tetrodotoxin abolished carbachol-induced release of PYY into lumen and vasculature. These data suggest that the ileal PYY release, into either lumen or vasculature, is under the control of postganglionic cholinergic neurons via muscarinic receptors. PMID- 7479340 TI - Biphasic effect of MCH on alpha-MSH release from the tilapia (Oreochromis mossambicus) pituitary. AB - The effect of melanin-concentrating hormone (MCH) on the release of alpha melanocyte stimulating hormone (alpha-MSH) from the tilapia pituitary gland was studied in vitro. In a superfusion set up, 10 nM to 1 microM synthetic salmon MCH caused a concentration-dependent inhibition of alpha-MSH release from tilapia neurointermediate lobes (NILs). Immunoneutralization of MCH in tilapia NILs further indicated that endogenous MCH has an inhibitory effect on the melanotropes. The release of monoacetylated alpha-MSH release was more strongly inhibited by MCH than that of des-, and diacetylated alpha-MSH, indicating that MCH modulates the secretory signal of the melanotropes in a quantitative and qualitative manner. A high concentration of MCH (10 microM) substantially increased the release of alpha-MSH. Further evidence in support of a stimulatory action of high concentrations of MCH was provided by the observation that the MCH analogue MCH(2-17) at 10 and 35 microM enhanced alpha-MSH release as well. Therefore, we conclude that the response of pituitary melanotropes to MCH is biphasic, as was reported previously for the effects of MCH on other targets in fish and mammals. Under physiological conditions the inhibitory action of MCH on fish melanotropes most likely dominates. PMID- 7479342 TI - Interaction of CCKB receptors with amphetamine in responding for conditioned rewards. AB - Cholecystokinin (CCK) has been localized in the nucleus accumbens (NAC) where it may interact with dopamine neurotransmission. NAC dopamine is involved in the control over behavior produced by conditioned rewards. The present experiment tested the whether blockade of endogenous CCKB receptors with L-365,260 (0.1 mg/kg, IP) potentiates bar pressing for stimuli previously associated with food reward. Intra-accumbens amphetamine (20 micrograms) facilitated bar pressing for conditioned rewards. Systemic administration of L-365,260 potentiated this amphetamine response but produced no effect on responding when administered alone. These findings suggest that endogenous CCKB mechanisms may normally inhibit DA function in reward-related behaviors. PMID- 7479343 TI - Changes in amino acid levels in rat plasma, cisternal cerebrospinal fluid, and brain tissue induced by intravenously infused arginine-vasopressin. AB - Circulating arginine-vasopressin (AVP) is known to reduce the blood-to-brain transfer of large neutral amino acids (AA). As a first step to examine whether the reduced uptake by brain endothelial cells is reflected in changes in large neutral amino acid levels of the extracellular fluid environment of cells within the nervous tissue, we measured the concentrations of amino acids in plasma, cerebrospinal fluid (CSF), and hippocampal tissue of rats before and after infusion of AVP (34 and 68 ng/min/kg, respectively) over the time period of 60 min. AA levels changed in all compartments investigated during both saline and AVP infusions. Whereas in the saline-infused controls changes in CSF AA levels paralleled those in plasma, this correlation was abolished by raising AVP concentrations. The effect of AVP was found to be i) dependent on the AA, ii) different with respect to direction and iii) magnitude of changes in AA levels, and iv) in some cases dose dependent. In summary, AVP infusion increased plasma levels of 10 AA, but decreased all 15 AA measured by some 30% in CSF. In contrast to CSF, levels of AA were slightly enhanced in the hippocampal tissue. The results are not solely explicable by a reduced blood-to-brain transfer of AA. We conclude that further mechanisms by which AVP affects the availability of AA to the brain may exist. The physiological significance of the findings might be related to brain osmoregulation, especially in situations of stress. PMID- 7479344 TI - Differential modulation of alpha 2-adrenergic and mu-opioid spinal antinociception by neuropeptide FF. AB - Neuropeptide FF (NPFF) has been found to act as an antiopioid peptide. However, IT NPFF has recently been shown to potentiate the antinociceptive effects of IT morphine and to produce antinociception on its own. The aim of this study was to find out whether pretreatment with NPFF causes a comparable potentiation of dexmedetomidine-induced antinociception. NPFF (0.05-10.0 nmol) produced no antinociceptive effects in the rat tail flick test. NPFF potentiated the antinociceptive effect of IT morphine (7.8 nmol). This potentiation was prevented by IT naltrindole (28 nmol), which did not attenuate the antinociceptive effect of morphine. Dexmedetomidine (1.6-6.4 nmol) produced a dose-dependent antinociceptive effect, which was not potentiated by NPFF. Activation of the endogenous delta-opioid system due to the antiopioid effect of IT NPFF is proposed as an explanation to the reported differential action of NPFF on the mu opioid and the alpha 2-adrenergic systems. PMID- 7479346 TI - The value of cytometric DNA analysis as a prognostic tool in neuroendocrine neoplastic diseases. AB - In several traditionally non-endocrine, common, human, neoplastic diseases, it has become well established during the last few years, that cytometric analyses of the DNA distribution pattern of the nuclei of tumour cells can be an excellent supplement to the conventional prognostic tools, (such as clinical staging and histopathologic malignancy assessments). When analogous studies of the value of DNA analysis by means of flow cytometry and/or image cytometry are made in neuroendocrine (NE) neoplastic diseases, the ensuing results often become rather disappointing. Thus, clear-cut aneuploid DNA histograms can be found in the neoplastic cell nuclei of clinically and histopathologically completely benign NE adenomas (and even hyperplastic nodules). In contrast, highly aggressive NE carcinomas not seldom reveal themselves to be composed of tumour cells with nuclei, displaying an euploid, i.e. normal, DNA pattern. Statements of this kind have been based on the results of comprehensive investigations in several laboratories, analysing such NE tumours as insulomas/insular carcinomas, bronchial/gastrointestinal carcinoids, phaeochromocytomas, paragangliomas, neuroblastomas, adenomas of the anterior pituitary gland, parathyroid adenomas, medullary carcinoma of the thyroid and Merkel-cell tumours of the skin. Thus, the prognostic value of the cytometric DNA ploidy pattern of the nuclei of neoplastic parenchymal cells is definitely lower in NE tumours than in most of the traditionally non-endocrine carcinomas and sarcomas. Data from published and unpublished series of these kinds of NE tumours, and those of prostatic and breast carcinomas with NE differentiation, are given. By means of a new, consecutive double staining technique, it was shown that in idiopathic nesidioblastosis, the hyperinsulinism is caused by beta cells with a nuclear DNA ploidy pattern of euploid type. By the same technique, it can be shown that in the pathogenesis of the hypergastrinaemia-induced ECL-cell carcinoids of the stomach, a switch from an euploid to an aneuploid nuclear DNA distribution pattern occurs in the ECL-cells when they pass from a state of hyperplasia to that of a genuine neoplasia. In neuroblastomas, a triploid (i.e. aneuploid) DNA pattern is part of an algorithm capable of predicting a 96% survival rate, whereas a diploid/tetraploid (i.e. euploid) DNA pattern predicts a 0% survival. PMID- 7479347 TI - Recent advances in cytometry, cytogenetics and molecular genetics of thyroid tumours and tumour-like lesions. AB - In this review we summarize the most recent advances of flow and image cytometry, cytogenetics and molecular genetics of tumours and tumour-like lesions of the thyroid, especially focusing on the putative relationship between nodular goitres and adenomas, the differential diagnosis of benign and malignant neoplasms and the pathogenesis and prognosis of carcinomas. Data obtained by in situ hybridization and immunohistochemical detection of proliferative markers, metal binding proteins and oncogenes, anti-oncogenes, growth factors and growth factor receptors are also used whenever appropriate. PMID- 7479345 TI - ACTH-related peptides: receptors and signal transduction systems involved in their neurotrophic and neuroprotective actions. AB - ACTH-related peptides are promising neurotrophic and neuroprotective agents, as demonstrated in many in vivo and in vitro studies. They accelerate nerve repair after injury, improving both sensor and motor function. Furthermore, ACTH-related peptides have neuroprotective properties against cisplatin- and taxol-induced neurotoxicity, they improve neuronal function in animals with neuropathy due to experimental diabetes, and they prevent degeneration of myelinated axons in rats suffering from experimental allergic neuritis, a model of peripheral demyelinating neuropathy. Studies in neuronal cultures have corroborated these clinical observations and serve to investigate the mechanism of action of the ACTH-related peptide effects. This paper reviews both in vitro and in vivo effects and emphasizes the mechanism of action. Recent data on melanotrophic receptors and signal transduction systems will be discussed in this context. PMID- 7479348 TI - Diagnostic problems in differentiated carcinomas of the thyroid. AB - In this review article we consecutively address the following issues: differential diagnosis of follicular adenomas and minimally invasive follicular carcinomas, with a special emphasis on the hyalinizing trabecular tumours; differential diagnosis of benign and malignant papillary lesions; individualization of the variants of papillary carcinoma that carry a guarded prognosis; differential diagnosis of differentiated thyroid carcinomas exhibiting foci of other types of thyroid carcinoma, namely mucoepidermoid and poorly differentiated carcinoma; and, finally, the diagnostic problems raised by the so called mixed medullary-follicular carcinomas. PMID- 7479349 TI - Diagnostic problems in medullary carcinoma of the thyroid. AB - Medullary carcinoma of the thyroid (MCT), the tumor entity related to parafollicular C cells of the thyroid gland, is characterized by peculiar morphological, functional and behavioral features. Apart from the constant finding of calcitonin production by MCT, heterogeneous patterns of growth as well as different associations with other lesions in the setting of MEN II syndrome have been observed. This review addresses these points, as were discussed at the 1992 European School of Pathology courses. Heterogeneity of histological patterns of MCT suggests caution in the interpretation of any thyroid tumor. Immunohistochemical demonstration of calcitonin is apparently the only valid criterion for a differential diagnosis. A correct identification of MCT is important for the individual patient (to plan proper therapy) but also for patient's relatives to screen other familiar cases in the setting of MEN II syndromes. PMID- 7479350 TI - Pituitary lesions in multiple endocrine neoplasia syndrome (MENS) type 1. AB - This review summarizes knowledge on pathology of proliferative lesions of the pituitary gland in multiple endocrine neoplasia syndrome type 1 (MENS 1). When compared with sporadic tumors occurring in general population, pituitary adenomas associated with MENS 1 show the following distinguishing features: (1) they are more often endocrinologically functional, (2) they are more often GH or PRL producing and (3) they are more frequently plurihormonal. The study of autoptic cases discloses that pituitary PRL or mixed GH-PRL cell adenomas in MENS are multiple and are associated with PRL or GH cell hyperplasia of the peritumoral parenchyma. PMID- 7479351 TI - Null cell adenomas and oncocytomas of the pituitary gland. AB - This review highlights the most interesting features of null cell adenomas and oncocytomas of the pituitary gland. Recently, application of sensitive methods have documented a very low amount of active hormone production in these clinically nonfunctioning tumours. However, further studies are needed to clarify the histogenesis of null cell adenomas and oncocytomas. PMID- 7479352 TI - Histopathological diagnosis of parathyroid diseases. AB - Hyperparathyroidism is the predominant disease of the parathyroid gland. This disease is nowadays quite common and is often diagnosed at an earlier stage, mainly by means of serum calcium determinations on wide indications. This means that when detected, the glandular abnormalities may be less advanced, which could hamper differentiation of adenoma from chief cell hyperplasia, and of normal glands from slightly hyperplastic ones. Normal glands and pathological glands both show wide variations in size, cellular composition and arrangement, as described in this article. The usefulness of applying fat staining to distinguish between normal and abnormal glands is also reported. It is important to bear in mind that the parathyroid diagnosis is in fact an indirect diagnosis based on the assessment of an associated gland or glands. PMID- 7479353 TI - Endocrine tumors of the small and large intestine. AB - Among endocrine tumors arising in the intestinal tract, midgut argentaffin EC cell carcinoids, duodenal gastrin cell tumors and rectal trabecular L cell carcinoids, in order of decreasing frequency, are those better represented. Together they account for more than 80% of such tumors. Duodenal somatostatin cell tumors, gangliocytic paragangliomas and poorly differentiated neuroendocrine carcinomas, are also well defined tumor entities. The carcinoid syndrome with intermittent flushing, hypotension and diarrhea, and the Zollinger-Ellison syndrome with severe peptic ulcer disease, are the only hyperfunctional syndromes consistently found in association with these tumors. The carcinoid syndrome arises in about 10% of intestinal carcinoids, usually in their advanced metastatic stage. The Zollinger-Ellison syndrome occurs in association with about 40% of gastrin cell tumors, including small intramural growths. Tumor prognosis depends on mode and site of presentation, histology, cell type(s), size, level of invasion, metastases (especially distant metastases) and associated clinical syndrome or background disease. PMID- 7479354 TI - Endocrine tumours of the stomach. AB - This review illustrates the current classification of gastric endocrine tumors: a) carcinoid tumors (associated with chromic atrophic gastritis; with Zollinger Ellison syndrome and multiple endocrine neoplasia type I; sporadic) and b) neuroendocrine carcinomas. The pathogenetic characteristics and clinical implications that justify this classification are discussed. PMID- 7479355 TI - Nucleolar and AgNOR-analysis of prostatic intraepithelial neoplasia (PIN), atypical adenomatous hyperplasia (AAH) and prostatic carcinoma. AB - Atypical glandular proliferations of the prostate are found in combination with prostate carcinomas but also with benign prostatic hyperplasia (BPH). These atypical glandular lesions that mimic glandular carcinoma within the anterocentral part of the prostate (transition zone) are named "atypical adenomatous hyperplasia (AAH)." The term for atypical ductal and acinar mainly papillary and/or cribriform proliferations in the peripheral zone is "prostatic intraepithelial neoplasia (PIN)." In this study the significance of the status of nucleoli and the number of silver-stained nucleolar organizing regions (AgNOR) as a method for measuring proliferative activity was investigated in biopsies of AAH, PIN and carcinomas of different grades of malignancy. The aim was to prove whether there are relationships of AAH to low grade carcinomas on the one hand and of PIN to high grade carcinomas on the other hand. The frequency of nuclei with small nucleoli was low in BPH (0.6%). The values increased to 8.3% in AAH, and 28.7% in PIN. The frequency of nuclei with prominent nucleoli in low grade carcinomas was 41.7% and in high grade carcinomas 81.1%. Usually, one single nucleolus was found per nucleus. Multiple prominent nucleoli were found in high grade carcinomas and in some cases of high grade PIN only. The result of AgNOR analysis was similar: low numbers in BPH, low to moderate values in AAH, moderate values in low grade carcinomas and high values in PIN, as well as high grade carcinomas. Besides correlation between topography, histology and cytology, the similarities of nucleolar and AgNOR-analysis between PIN and high grade carcinomas support the idea that PIN is the precursor of the peripheral prostatic carcinoma. The histological, cytological and cell kinetical features of AAH are intermediate between BPH and low grade carcinoma of the prostate. Therefore, the findings to date are inconclusive and further follow up studies are necessary to prove if AAH may be a precursor of transition zone carcinoma of the prostate. PMID- 7479356 TI - The expression and localization of bcl-2 protein in normal skin and in non melanoma skin cancers. AB - Non-melanoma skin cancers are the most commonly diagnosed malignancies and are typically indolent in their clinical behavior. Although predisposing factors leading to the development of these cancers, such as ultraviolet irradiation, are well described, the molecular events involved in their pathogenesis are incompletely understood. The localization of bcl-2 expression within the skin was determined using immunohistochemical methodologies and an anti-bcl-2 monoclonal antibody. The cytoarchitectural distribution of bcl-2 protein in normal skin included basal keratinocytes, the dermal papillae of the hair follicle, the keratinized Huxley's and Henle's layers, and the keratinized outer root sheath cells of the isthmus and infundibulum of the hair follicle. In addition, intense immunoreactivity was noted in the secretory coil of eccrine sweat glands. The distribution of bcl-2 protein within normal skin did not correlate with the known histologic localization of stem cell compartments. Basal cell carcinomas expressed high levels of bcl-2 protein. In contrast, squamous cell carcinomas typically exhibited no immunohistochemically detectable bcl-2 protein. The findings suggest a potential contribution of bcl-2 gene deregulation to the pathogenesis of some types of non-melanoma skin cancer. PMID- 7479357 TI - Detection of loss of heterozygosity in the p53 tumor suppressor gene using a PCR based assay. AB - Inactivation of the p53 tumor suppressor gene has been reported to be a prognostic factor in several human cancer types. Normal function of the gene is affected by deletion in one allele; dysfunction of the other allele is often caused by a mutation. In tumors of heterozygous individuals, deletion of one allele can be detected as loss of heterozygosity (LOH). A recently found variable number of tandem repeats (VNTR) segment in intron 1 of the p53 gene seems to be highly polymorphic and, therefore, a very useful marker in detecting LOH in various types of tumor samples. We in vitro amplified the VNTR segment from genomic DNA samples of 101 lung cancer patients and run conventional agarose gel electrophoreses in order to detect the alleles of various length, differing by the number of repeats. The usefulness of the method was studied using DNA from white blood cell samples and from fresh and formalin-fixed, paraffin-embedded tumor samples. Of the patients, 56% were found to have two different alleles, i.e. were informative in this assay. In 18% of the lung tumors from the informative cases, LOH in the p53 suppressor gene was detected. PMID- 7479358 TI - Urokinase plasminogen activator induces angiogenesis and tumor vessel invasion in breast cancer. AB - Urokinase plasminogen activator (uPA) is a proteolytic enzyme implicated in cancer invasion and tumor progression. Urokinase PA and its inhibitor (PAI-1) appear to be new and independent prognostic markers in breast cancer. To investigate how uPA- and PAI-1-levels correlate with angiogenesis and tumor vessel invasion, we counted microvessels and their tumor invasion and determined the uPA- and PAI-1 levels in 42 primary invasive breast carcinomas. 20 Patients had no lymph node metastasis at the time of surgery, while 22 patients had positive nodes. Using light microscopy, we highlighted the vessels by staining their endothelial cells immunocytochemically for CD31 and Factor VIII. After gaining tumor tissue extracts, we determined the uPA- and PAI-1-levels by ELISA. A positive correlation between microvessel density, angioinvasion and uPA- and PAI-1-levels was found. We speculate that high uPA levels may induce tumor neovascularisation, angioinvasion and may cause tumor progression and metastasis. The degradation of the vessel wall by uPA causes a leak. This wall defect may, on the one hand, be the stimulus for endothelial cell proliferation and formation of new blood vessels and, on the other hand, it may be the place of tumor cell entry. PMID- 7479360 TI - Double duplication in a nonrotational colon. Study of a case associated with mucinous adenoma. AB - Duplications of gastrointestinal tract are unusual anomalies, especially in the colon. We report a case with two colonic duplications in a 20 year old woman who complained of recurrent backache associated with left lower quadrant abdominal pain. Neither physical examination nor laboratory test alterations were detected. Roentgenographic examination showed organic scolinosis, no intestinal gas and a mass with central hyperdensity and thickened wall in the descending colon area on the CT scan. Laparotomy was performed in which 22 cm of ascending colon and terminal ileum sited on left side, was excised. Pathological study revealed two unrelated and cystic duplications without communication to intestinal lumen. Their epithelial lining were of colonic or gastric types and showed a mucinous adenoma; the remaining layers were normal except for eosinophilic inflammatory infiltrate with Charcot-Leyden like crystaloids. In conclusion, we present a double spherical colonic duplication type I associated to a mucinous adenoma on a non-rotational ascending colon in a patient with organic scoliosis. PMID- 7479359 TI - The use of CD31 and collagen IV as vascular markers. A study of 56 vascular lesions. AB - A monoclonal antibody against the endothelial cell adhesion molecule CD31 (endo CAM, PECAM-1) was tested on wax embedded tissue of 56 cases of vascular lesions including benign and malignant vascular neoplasms. Additional preparations were stained with antibodies to type IV collagen and to von Willebrand factor/Factor VIII-RAG (vWf). Our results prove reliability of CD31 and its superiority to vWf in the labelling of endothelium and its subsets in such lesions, with the exception of lymphangiomas. As markers for basal lamina collagen and endothelium respectively, type IV collagen and CD31 together provide a powerful tool for 1) the diagnosis of endothelial neoplasms and 2) the definition of endothelial, mural and pericytic or perivascular tissue compartments in vascular lesions of complex architecture. Hence, we use CD31 and type IV collagen in cases of presumed vascular neoplasms, adding other markers to the panel in accordance with the differential diagnosis, as well as in the recognition of compromised endothelia, such as in vascular invasion by various malignant neoplasms. PMID- 7479361 TI - Degradation of macromolecules during preservation of lyophilized pathological tissues. AB - Methods of preserving biological specimens are becoming more important due to recent advances in molecular biological analysis. Storing samples in a freezer, however, is still the most commonly used method of preserving pathological specimens. We investigated the feasibility of using freeze-dried tissues stored at room temperature as an alternative method of preserving tissue samples for molecular analysis at the DNA, RNA, and protein levels. The freeze-dried tissues were transferred to a sealed vial filled with nitrogen gas and kept for months at room temperature. DNA and protein were stably preserved for at least 24 weeks. RNA, however, showed slight degradation after 10 weeks of storage. Controlling moisture and temperature during long-term storage was found to be important, as it significantly affected the stability of these cellular molecules in tissues. Shelf-stable preservation eliminates the need for storage in a freezer and allows convenient shipping of samples to distant places. These findings should provide practical basis for the development of a convenient and economical way of preserving pathological specimens for a variety of analyses in the field of molecular biology. PMID- 7479362 TI - The neuropathology and epidemiology of AIDS. A Berlin experience. A review of 200 cases. AB - The brains of 200 patients who died with Acquired Immunodeficiency Syndrome (AIDS) from Berlin were examined retrospectively. This study was specifically intended to evaluate and document the prevalence of neuropathologic abnormalities, establishing the frequency of the various types of structural lesions, their combinations, their relative incidence, and the risk factors involved in different age groups. The data were compared and contrasted with the findings reported from other parts of the world and other German cities. It was found that the mean age of this group of patients was 41.4 years old, 75% were homosexual/bisexuals (H/B) and 18.5% were drug abusers (DA). Only 5.5% were women. Brain parenchymal changes, called in this report, HIV-related encephalopathy (HIVRE), characterized by vacuolization or spongy changes and astrocytosis in the subcortical white matter, and occasionally in gray matter, were found in 67 patients (33.5%). Drug abusers had a higher incidence of HIVRE (59.5%) compared with homosexual/bisexuals (28%). This is statistically significant (p < 0.0005). CMV encephalitis was found in 26 patients (13%) (8% of the drug abusers in contrast to 13% in the homosexual/bisexuals group). Primary central nervous system lymphoma (PCNSL) was seen in 28 patients (14%) regardless of the risk factor involved. 20 (13%) of the 150 H/B and 3 (8%) of the 37 DA had CMV encephalitis. Of the 150 H/B, 24 (16%) had PCNSL compared with only 4 of 37 (11%) of the DA. A significant incidence of opportunistic infections, both protozoal and viral was found in all groups. Cerebral toxoplasmosis occurred in 68 patients (34%). Microglial (phagocytic) nodules, probably related to CMV or cerebral Toxoplasmosis, were observed in 40 cases (20%). Diffuse microglial proliferation was noted in 104 patients (52%). Cerebral cryptococcosis was found in three patients. Progressive multifocal leukoencephalopathy was seen in 16 patients (8%). Various combinations of CNS pathological processes were found in 44 of the patients (22%). These include concomitant infections with Toxoplasma gondii and HIVRE in 13 patients; Toxoplasmosis and PCNSL in 8 patients; Toxoplasmosis with CMV and HIVRE in 4 patients; Toxoplasmosis with CMV in 2 patients; Toxoplasmosis with PCNSL and CMV in 2 patients; Toxoplasmosis with PCNSL and HIVRE in 2 patients and Toxoplasmosis with PML and HIVRE in 2 patients; Cerebral CMV with PCNSL and HIVRE in 4 patients; Cerebral CMV with HIVRE in 2 patients; PML with PCNSL in one patient; PML with HIVRE in 2 patients; and PML with PCNSL and HIVRE in one patient. Cerebrovascular lesions were found in 34 patients (17%).(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 7479363 TI - Genotype study of non-Hodgkin's lymphoma. AB - DNA from 47 patients with non-Hodgkin's lymphoma (NHL) was studied for immunoglobulin and T-cell receptor (TCR) gene rearrangements with Southern blot hybridization. In 83% of the cases the genotypic changes were consistent with immunophenotypic and morphologic examination. Two cases showed mixed genotype and 9 cases of B-cell NHL (67% of centroblastic, 36% of follicular and 33% of large cell anaplastic) showed a population of cells with TCR gamma rearrangements in addition to immunoglobulin rearranged bands. We compared the TCR gamma variable region usage in these rearrangements in B-cell NHL with T-cell NHL and reactive hyperplasia. In T-cell NHL TCR gamma variable regions located at the 3' part of the variable locus were used more often, whilst in B-cell NHL regions of the 5' portion of the locus were preferentially used. Our results confirm the genotypic heterogeneity of histologically defined subtypes of NHL. PMID- 7479364 TI - Multiple skeletal muscle metastases from malignant pleural mesothelioma. AB - A 54-year-old male patient (occupational asbestos exposure over 27 years) died 2 years and 4 months after the diagnosis of malignant pleural mesothelioma of the right side, despite twice undergoing pleurectomy and radiotherapy. The autopsy revealed a locally advanced pleural mesothelioma of both sides involving the pericardium, heart, right diaphragm, liver and peritoneum. Disseminated metastases in numerous lymph nodes and a hematogenous metastatic spread into both lungs, the thyroid gland, peritoneum and skeletal musculature were found. The left temporal muscle and proximal limb skeletal muscles of the right upper arm and both thighs exhibited multiple metastases measuring up to 7 cm in diameter. Microscopically, a biphasic type of mesothelioma was detected. To our knowledge this is the first extensive case report on muscle metastases in malignant mesothelioma. Reviewing the literature, it is thought that in the presented case the long survival time of the patient and his protracted preterminal immobility could have contributed to the unusual formation of multiple skeletal muscle metastases. PMID- 7479365 TI - A case of pleomorphic T-cell lymphoma with a high content of reactive histiocytes presented with hypereosinophilia. AB - A case of peripheral T-cell lymphoma classified, according to the updated Kiel classification, as a large pleomorphic T-cell lymphoma with a high content of reactive histiocytes and blood hypereosinophilia is reported. Light microscopic examination revealed a diffuse effacement of the lymph node structure by large pleomorphic lymphoma cells mixed with eosinophils and many histiocytes, some of them presenting discrete features of hemophagocytosis. The neoplastic cells were CD3, CD5, CD8 and HLA-DR positive but failed to show CD30 antigen. DNA molecular analysis displayed simultaneous rearrangements of the genes coding for the delta chain of the T-cell receptor and for the Ig heavy chain. Increased serum levels of angiotensin converting enzyme and ferritin were found and probably induced by the reactive histiocytes. Immunoassays (ELISA) with antibodies directed against some cytokines and against the Tac peptide (sIL-2R) were performed. They demonstrated high serum levels of sIL-2R and a slight increase in GM-CSF, but neither IL-5 nor IL-3. The association of blood hypereosinophilia and histiocytic hyperplasia with a peripheral T-cell lymphoma is discussed. PMID- 7479366 TI - Primary osteosarcoma of the breast. AB - A case of primary osteosarcoma of the breast in an 83 year-old woman was studied by using a large immunohistochemical panel of antibodies. Tumor cells were stained intensely for vimentin, and focally for alpha-1-antitrypsin. No immunoreactivity was detected for cytokeratin. The absence of an epithelial component excludes a metaplastic carcinoma. The multinucleated giant cells stained strongly for KP-1 antibody, indicating a histiocytic nature. The question is raised whether osteosarcoma of the breast is a malignant tumor de novo, or develops by transformation of a fibroadenoma. PMID- 7479367 TI - Establishment of two rat osteosarcoma cell lines (YROS-1 and YROS-2) induced by radioactive phosphorus. AB - Two rat osteosarcoma cell lines, YROS-1 and YROS-2, were established from two experimental osteosarcomas and induced by internal irradiation with radioactive phosphorus. Both cell lines formed a monolayer cell sheet in vitro with focal piling. The YROS-1 cells were refractile and spindle or polygonal in shape, whereas the YROS-2 cells were flat, spread and polygonal in shape. Ultrastructurally, the YROS-1 cells had well-developed rough-surfaced endoplasmic reticulum with focal pericellular deposition of calcified matrix, whereas YROS-2 had abundant polysomes and intracytoplasmic filaments. Both cell lines grew stably with population doubling times of 23 and 39 h, respectively. Flow cytometry revealed that YROS-1 was rich in proliferating cells compared to YROS 2, with a higher colony-forming efficiency. YROS-1 showed high alkaline phosphatase activity, while YROS-2 possessed low activity. When subcutaneously transplanted into lumbodorsal area of athymic nude mice, only YROS-1 formed tumors with frequent lung metastasis. PMID- 7479368 TI - Pierre Dustin (1914-1993), founder of the European Society of Pathology. PMID- 7479369 TI - Distribution and expression of the alpha-tubulin mRNA in the hippocampus and the temporal cortex in Alzheimer's disease. AB - The distribution of the messenger RNA for alpha-tubulin has been investigated by in situ hybridization in the human hippocampus and temporal cortex in normal subjects and in Alzheimer's disease. The alpha-tubulin mRNA was strongly expressed in neurons in the gyrus dentatus, in the Ammon's horn and in cortical layers of the temporal cortex. The same distribution was observed in Alzheimer's disease. An important reduction of the hybridization signal was apparent, however, in areas rich in neurofibrillary lesions, e.g. as in layer II of entorhinal cortex. Neurons containing neurofibrillary tangles exhibited a weaker hybridization signal than adjacent neurons devoid of neurofibrillary tangles. The immunoreactivity for alpha-tubulin was drastically reduced in tangles-bearing neurons. These results indicate that tubulin transcription is reduced in tangles bearing neurons, a reduction which might play a role in the reported decrease in the number of microtubules in neurons containing neurofibrillary tangles. PMID- 7479370 TI - Histopathology and ultrastructure of the thymus in OP/OP rats with special focus on thymic macrophages. Effect of bone marrow transplantation. AB - The thymi of op/op rats were investigated before and after bone marrow graft. Normal littermates were used as controls. The thymic involution and atrophy in non-treated op/op rats was characterized by the presence of pyknotic thymocytes (apoptosis) and numerous lipid-laden macrophages containing pleiomorphic lysosomes. This thymic atrophy is histologically very similar to the thymic involution induced by glucocorticoids or by stress in normal rats. In treated new born op/op rats, the osteopetrotic process began to be reversed 2 weeks after the graft. The thymus of treated op/op recovered a normal weight and its histology and ultrastructure were quite similar to that of normal littermates. These findings are discussed in relation to the defective macrophage function and cellular immunity impairment reported in osteopetrosis. PMID- 7479371 TI - Morphological changes of thymus in retrovirus-induced murine acquired immunodeficiency syndrome (MAIDS). AB - The possible contribution of the thymus in the setting of acquired immunodeficiencies is still questioned. Here we report some new findings regarding a potential involvement of the thymus in mice infected with RadLV-Rs, a viral mixture inducing murine acquired immunodeficiency syndrome (MAIDS). Thymi were sequentially removed, weighted and morphologically analyzed at different time intervals post-infection. Infection with RadLV-Rs led to a decrease in thymus weight mostly apparent from the fourth week. The first changes were seen at the third week as perivascular clusters of B-cells at the cortico-medullary junction. The ensuing process of atrophy mainly involved the cortex, while a mixed population of large T- and B-cells filled the medulla. These observations are discussed with regard to the pathological changes occurring in other lymphoid and non-lymphoid organs, in the context of the lymphoproliferation and immunodeficiency characterizing the disease, and by comparison with other models of retrovirus-induced immunodeficiencies. PMID- 7479372 TI - Ductular reaction in the liver. PMID- 7479373 TI - Primary non-Hodgkin lymphoma of the gastric stump. AB - Patients with a history of gastroduodenal surgery for benign lesions are at increased risk of developing cancer of the gastric stump. The majority of malignant tumors arising in gastric remnants are adenocarcinomas, but a few of cases of malignant lymphoma have been reported as well. Some recent studies have suggested that Helicobacter pylori-associated gastritis may play a role in the pathogenesis of primary gastric lymphoma. We report a case of primary high grade polymorphic B-cell lymphoma (high grade MALT lymphoma) occurring in the gastric stump of a 74-year-old man 17 years after Billroth-II gastrectomy. The non tumorous gastric mucosa showed features of reflux gastritis with presence of deeply situated dense lymphoid aggregates in the lamina propria. However, no micro-organisms with the morphology of Helicobacter pylori could be found at the luminal surface. The literature concerning gastric stump lymphomas is reviewed. PMID- 7479374 TI - Distribution of interferon-gamma receptors in normal and psoriatic skin. AB - Recent data suggest that imbalances in production and secretion of cytokines, in particular interferon-gamma (IFN-gamma), may be crucial in the pathogenesis of psoriasis. In order to exert its role on target cells, IFN-gamma has to interact with a specific cell membrane receptor termed the IFN-gamma-receptor (IFN-gamma R). We studied the distribution of IFN-gamma Rs in frozen skin biopsies from 25 psoriatics and 5 normal controls with two unrelated monoclonal antibodies, and compared its distribution with that of the IFN-gamma-inducible HLADR- and ICAM-1 antigens. In normal skin, IFN-gamma Rs were restricted to the basal cell layer; weak staining was found on scattered mononuclear cells in the papillary dermis. In 13/25 active psoriatic lesions, additional suprabasal immunoreactive foci, and in 5/25 cases, diffuse immunoreactivity of the entire epidermis were seen. No striking topographical similarities between the site and number of IFN-gamma R+, HLADR+ and ICAM-1+ keratinocyte foci were observed, suggesting that cytokines other than IFN-gamma induce HLADR-antigens on psoriatic keratinocytes in vivo. The restricted distribution of IFN-gamma R on the germinative cell layer in normal skin confirms the role played by IFN-gamma in the normal growth regulation of the epidermis. The de novo suprabasal expression of IFN-gamma R in psoriasis argues against the current hypothesis that IFN-gamma R are down-regulated due to a local excess of IFN-gamma or transforming growth factor alpha (TGF-alpha). Whether IFN-gamma Rs in psoriatic skin are functionally normal and involved in signal transmission, remains to be studied. PMID- 7479375 TI - Unusual intracytoplasmic inclusions in metastatic carcinoma. Discussion of their possible significance. AB - Unusual filamentous inclusion bodies in the cytoplasm of metastatic tumor cells are described. Their presence (intermingled with zymogen granules) seems rather restricted to cells of primary or metastatic acinar cell carcinoma of the pancreas, acinic carcinoma of salivary gland (parotid gland) and Paneth cells (neoplastic or in zinc deficiency state). For the time being, the real nature of these inclusions (deranged zymogen granules?) is unclear. This case also emphasizes the value of electron microscopy in solving the problem of the occult primary tumor and avoiding the misdiagnosis of an endocrine tumor (e.g. islet cell tumor or carcinoid), or a duct cell tumor with eosinophilic granular cytoplasm or in recognizing the foci of acinar cells in a mixed variant of carcinoma of the pancreas. PMID- 7479376 TI - Development. The science of damage limitation. AB - The building of a normal embryo is not accomplished by following a series of complex instructions; rather a number of critical processes enable the cells involved in the process of development to follow pathways which subsequently constrain their growth, mobility and differentiation. There is considerable redundancy in the system, both in terms of mechanisms and permissible outcomes. The way that these processes have become established in evolution gives us a number of insights into how these processes may fail and information from developmental biology informs us about a number of other pathological processes. PMID- 7479377 TI - Radial scar versus tubular carcinoma of the breast. A comparative study with quantitative techniques (morphometry, image- and flow cytometry). AB - The present study is focused on the differential diagnosis between radial scar (RS) and tubular carcinoma (TC) using morphometrical and cytophotometrical analysis (static and flow cytometry) of a number of histologically well established RS cases (17 lesions) compared with 6 early infiltrating small TC with sclerotic stroma and pseudo-RS fields. One case displayed both RS and TC foci in contiguity. Mean nuclear area was larger in the group of tubular carcinomas (51.0 mu 2) than in the case of radial sclerosis (38.30 mu 2). We also found a larger number of aneuploid cases in tubular carcinomas measured by image cytometry, but both types of lesions were diploid when measured by flow cytometry; only one case of radial scar resulted aneuploid. PMID- 7479378 TI - Lymph node involvement revealing a lymphomatous polyposis of the gastrointestinal tract. AB - The digestive tract is the most frequent site of extranodal malignant lymphomas. Lymphomatous polyposis is one of them, and its prognosis is poor. It corresponds to a digestive localization of mantle cell lymphoma. In most cases it is discovered following digestive symptoms. However, in some cases this digestive malignant lymphoma may be asymptomatic. Thus complete endoscopic exploration of the digestive tract including biopsies is necessary for every patient presented with lymph node mantle cell lymphoma. PMID- 7479379 TI - New aspects of cell biology in osteosarcoma. AB - Among the solid tumors of childhood and adolescence, osteosarcoma (OS) represents the most prominent example of efficient aggressive chemotherapy with secondary surgical therapy. A specific subclassification of the tumor is indispensable and must include recent results of cell biology. The co-distribution of different collagen types I-VI reflects the diverse differentiation of osteosarcoma cells, supporting the concept of a pluripotent mesenchymal cell to be the stem cell of the tumor. In contrast, osteonectin (SPARC) may not be considered as a reliable marker for osteosarcoma. The experience of special proteins being secreted by osteosarcoma cells is rather limited. Detailed molecular biological studies are still lacking. A loss of alleles on chromosome 17, particularly in the defined region 17p 13, can be observed in more than 75% of all OS, suggesting the contribution of a tumor suppressor gene, p53, located in that region. It is a 53 kd nucleophosphoprotein binding the major transforming protein, the large T antigen of Simian Virus 40. Immunohistological results showed positive staining with the antibody Pab 240 in 13 of 18 cases. In one osteoblastic OS, a novel splice mutation resulting in a fusing of exon 5 directly to exon 7 was detected. RB1 gene is also of major importance for the tumorigenesis of OS. The multidrug resistance (mdr) is associated with a membrane-bound channel-forming transport protein, the P-glycoprotein. It is a conserved plasma membrane component of about 170 kd. Both the human isoforms mdr 1 and mdr 3 are localised in the long arm of chromosome 7.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7479380 TI - New elements for an updated classification of the carcinomas of the stomach. AB - Based upon the results of a thorough study of 213 patients submitted to potentially curative resection we propose the following histologic classification of gastric carcinoma: isolated-cell carcinoma (6.6%), glandular carcinoma (41.8%), solid carcinoma (13.1%) and mixed carcinoma (38.5%). Half of the mixed carcinomas displayed a predominant isolated-cell pattern, whereas in the other half the isolated-cells were a minor component (the predominant component being either glandular, solid, or both). The survival of patients with mixed carcinomas is significantly worse than those of patients with other histologic types of gastric carcinoma regardless of the site of tumors and the inclusion, or not, in the series, of post-operative deaths. The proposed classification keeps its independent prognostic significance in a multifactorial analysis, appearing as the second most important prognostic factor in patients with gastric carcinomas, after the TNM staging and before venous invasion. PMID- 7479381 TI - Immunotactoid glomerulopathy with unusually thick extracellular microtubules and nodular glomerulosclerosis in a diabetic patient. AB - It has recently been suggested that immunotactoid glomerulopathy be separated from much more common fibrillary glomerulonephritis by ultrastructural features of highly organized immune deposits containing tubules of more than 30 nm in diameter. We report and discuss the results of a light, immunofluorescence and electron microscopic study of a needle renal biopsy from a 75-year-old, non insulin dependant diabetic female presented with nephrotic syndrome, hypertension and a progressive renal failure. A unique coexistence of nodular glomerulosclerosis, as traditionally ascribed to diabetes with a peculiar type of immunotactoid glomerulopathy was confirmed by the exclusion of amyloidosis, monoclonal gammopathies, systemic autoimmune diseases and cryoglobulinemia. Mesangial, scattered subepithelial and segmentally prominent subendothelial immune deposits were found highly organized in mostly parallel arrays of 40 to 91 nm thick tubules. The average thickness of 67 nm exceeds the average diameter of tubules in all other 11 published cases of immunotactoid glomerulopathy to date. By immunofluorescence, predominantly capillary wall, thick, ribbon-like glomerular deposits contained IgG, IgM, kappa and lambda light chains of equal intensity, C3, C4 and fibrin related antigens. Mild to moderate glomerular cell proliferation associated with nodular sclerosis has been assumed to be causally related to immunotactoid deposits. PMID- 7479383 TI - XVth European Congress of Pathology. Copenhagen, Denmark, September 3-8, 1995. Abstracts. PMID- 7479382 TI - Pathological anatomy in Denmark, a historical survey. PMID- 7479384 TI - Diversity and modulation of plasminogen activator activity in human prostate carcinoma cell lines. AB - Baseline cellular plasminogen activator (PA) activity, and the cellular proteins responsible for variations in PA activity were evaluated in three human prostate carcinoma cell lines. Net PA activity in the cell lines PC-3, DU-145, and LNCaP was measured using a plasminogen-dependent fibrin lysis assay. These three cell lines were then analyzed to determine the specific protein(s) responsible for differences in PA activity. mRNA and protein levels of cellular urinary PA (uPA), tissue PA (tPA), PA inhibitor 1 (PAI1), PA inhibitor 2 (PAI2), and uPA receptor (uPAr) were measured using Northern analysis and ELISA assays. Net cellular PA activity in the three cell lines varied over a 3-fold range (PC3 > DU145 >> LNCaP). Net PA activity in the fibrinolysis assay demonstrated a direct correlation with mRNA transcript levels of uPA, tPA, PAI1, and uPAr (PC-3 > DU 145 > LNCaP). uPA protein was identified in both the PC-3 and the DU-145 lines. tPA, PAI1, and PAI2 proteins were identified only in PC-3 cells. In general, cellular protein levels correlated with mRNA levels. These findings demonstrate that prostate carcinoma cell lines vary in their net PA activity. This variability results from both qualitative and quantitative differences in the cellular expression of PA regulatory proteins. PMID- 7479385 TI - Molecular mass and carbohydrate structure of prostate specific antigen: studies for establishment of an international PSA standard. AB - Despite the widely accepted use of prostate specific antigen (PSA) as a marker of prostate cancer, this molecule has not yet been completely characterized. Past studies have well established, however, using both amino acid and cDNA sequencing techniques, that PSA contains 237 amino acids, with a molecular mass of 26,079 Da for the peptide moiety of the molecule. The present study reports analysis of this protein by ion spray mass spectrometry (ISMS) and analysis of its carbohydrate moiety by NMR spectroscopy. The predominant PSA molecular species detected by ISMS was at relative molecular mass (M(r)) of 28,430, indicating that PSA contains a carbohydrate residue of M(r) 2,351, for a total percentage of carbohydrate of 8.3%. Analysis of PSA by SDS-PAGE, however, showed a M(r) of 32,000 to 33,000, suggesting an overestimation of the molecular weight by the latter technique. The complete primary structure of the PSA carbohydrate chain was determined by NMR spectroscopy in combination with carbohydrate composition analysis. The experimentally determined carbohydrate content of PSA confirms that only one N-glycosylation site is occupied in the protein. The proposed carbohydrate structure is a diantennary N-linked oligosaccharide of the N acetyllactosamine type, with a sialic acid group at the end of each of the two branches. In addition, our data indicate that approximately 70% of the PSA molecules contain a fucose group in the core chitobiose moiety. The calculated molecular weight of this carbohydrate structure (M(r) 2,351.8) is in excellent agreement with the predicted molecular weight of the carbohydrate group, based on the M(r) 28,430 for PSA measured by ion spray mass spectrometry and M(r) 26,079 calculated from the consensus sequence for the peptide portion of the molecule. ISMS of PSA is thus proposed as a convenient and reliable method of quality control, an indispensible step towards international standardization of this very important tumor marker for detection and monitoring of prostatic diseases, especially prostate cancer. PMID- 7479386 TI - Identity of PSA purified from seminal fluid by different methods: comparison by amino acid analysis and assigned extinction coefficients. AB - To determine the true extinction coefficient of prostate specific antigen (PSA) and to measure any differences in PSA when isolated from seminal fluid by four different published methods, we studied 10 different lots of PSA by quantitative amino acid analysis. Despite an expected PSA concentration of 1 mg/ml based on gravimetric analysis at an average optical density of 1.45 at 280 nm, we recovered only 0.79 mg/ml by quantitative amino acid analysis (range 0.752 to 0.820 mg/ml with a coefficient of variation [C.V.] of 3.3% among the 10 lots). The concentration of 0.79 mg/ml was based on a molecular weight of 28,430 daltons for glycosylated PSA determined by ion spray mass spectroscopy [Belanger et al: Prostate, 27:187-197, 1995]. From these 10 amino acid analyses, we calculated the extinction coefficient of PSA at 280 nm as 1.84 +/- 0.04 ml x mg-1 x cm-1 (range 1.78 to 1.90 with a C.V. of 2.2%). Similar concentrations of PSA were obtained by amino acid analysis regardless of the method of purification. These observations support the presence of a single form of PSA in seminal fluid and are consistent with the molecular evidence that PSA is transcribed from a single gene locus on the long arm of chromosome 19 [Riegman et al.: Genomics 14:6-11, 1992]. They do not support the recent contention by the Roswell Park group that the PSA they isolated in 1979 [Wang et al.: Invest Urol 17:159-163, 1979; Wang et al.: Prostate 24:107-108, 1994] is different from p30 reported a year earlier [Sensabaugh: J Forensic Sci 23:106-115, 1978]. PMID- 7479388 TI - Serum free prostate specific antigen: isoenzymes in benign hyperplasia and cancer of the prostate. AB - Prostate specific antigen (PSA) in serum of patients with benign prostatic hyperplasia (BPH) or prostate cancer (P-CA) not bound to alpha-1-antichymotrypsin (ACT) was analyzed by chromatofocusing. The procedure allowed the simultaneous separation of complexed and free PSA and the fractionation of the free PSA fraction into several isoenzymes. The detection of the isoenzymes was strongly dependent on the combination of antibodies introduced in the applied commercially available immunoassays (Cobas Core, Delfia). Isoenzymes in sera of patients with benign prostatic hyperplasia were mainly situated in the pI range of 6.6 to 7.3. Isoenzymes in sera of prostate cancer patients or in PSA from LNCAP cells were mainly situated in the pI range 7.0 to 8.3. Neuraminidase treatment of the sera shifted the isoelectric points of all three sources towards more basic pHs. An irregular glycosylation process in the dysplastic cells of the prostate is suggested to be the cause for the shift of the isoelectric points. The difference of isoenzyme distribution along the pH axis is discussed as a diagnostic tool to differentiate between BPH and P-CA. PMID- 7479387 TI - Protein kinase C regulation of the adenylyl cyclase system in rat prostatic epithelium. AB - In the context of the crosstalk between transmembrane signalling pathways, we studied the loci within the stimulatory receptor/Gs protein/adenylyl cyclase system at which protein kinase C (PKC) exerts regulatory effects in rat prostatic epithelial cells. The treatment of cells with the PKC activator phorbol 12 myristate 13-acetate (PMA) resulted in an impairment of the stimulation of adenylyl cyclase activity in terms of both potency, as seen with both vasoactive intestinal peptide (VIP) and pituitary adenylyl cyclase-activating peptide (PACAP 27), and efficacy, as seen with the beta-adrenergic agonist isoproterenol. This inhibitory effect of PMA could be prevented by cell incubation with pertussis toxin but not with cholera toxin, pointing to a Gi- but not Gs-dependent mechanism. This hypothesis was reinforced by ADP-ribosylation experiments that showed a low extent of alpha i with pertussis toxin but no change of alpha s with cholera toxin, as well as by the observation of the loss of the ability of low Gpp[NH]p doses to inhibit forskolin-stimulated adenylyl cyclase activity (a measure of Gi function) after cell treatment with PMA. However, the phorbol ester did not modify the adenylyl cyclase catalytic subunit, as shown by experiments on direct stimulation of the enzyme by forskolin. Whatever the exact mechanisms, the results support a crosstalk between the PKC and the adenylyl cyclase systems in rat prostatic epithelial cells in terms of an impairment of adenylyl cyclase stimulation, due presumably to phosphorylation of both membrane receptors (coupled to Gs) and Gi protein, but not of Gs protein or the adenylyl cyclase itself. PMID- 7479390 TI - Commentary on maximal androgen blockade in prostate cancer: a theory to put into practice? AB - The population at risk of prostate cancer is on the increase, and so is public awareness of this disease. There has been an unresolved controversy surrounding the benefits of maximal androgen blockade (MAB) as a valid approach to treatment of non-curative prostate cancer since it was first proposed in 1945. How are we to interpret the data on MAB in order to give each patient the best advice on treatment? Studies of MAB using medical castration (luteinizing hormone-releasing hormone [LHRH] analogue plus antiandrogen) vs. LHRH analogues alone are inconclusive when viewed collectively, although the largest showed objective benefits for MAB. The remaining studies have insufficient power to show the expected effect size. Studies of MAB using surgical castration plus antiandrogen vs. surgical castration alone also gave inconsistent results, although a meta analysis is in favor of MAB on objective criteria of response. Among trials of MAB using an LHRH analogue vs. surgical castration alone, one is positive and the remaining two are neutral for MAB. No study shows MAB to be worse than either medical or surgical castration alone. An overall meta-analysis shows a trend for benefit with MAB but is not statistically significant. The existing data have strongly suggested that there may be a particular benefit for certain subgroups of patients (including those with minimal disease) but numbers studied have been too small to allow valid conclusions. The INT 0105 trial in progress may permit firmer conclusions to be drawn on this and other questions. In the meantime one of the drawbacks to current MAB regimens is the exchange of modest clinical advantages for the side effects of nilutamide and flutamide. Given that the disease is noncurative, improved quality of life is the main goal of therapy, and excellent tolerability of treatment is fundamental to this. In a comparative trial, bicalutamide (Casodex) was more effective than flutamide (each in combination with an LHRH analogue) in terms of time to treatment failure and produced a significantly lower incidence of diarrhoea. In conclusion, the evidence supports early use of adequate hormonal treatment, and this should mean either medical or surgical castration, ideally augmented by an antiandrogen. Tolerability of the antiandrogen is a key consideration in gaining an improvement in quality of life with MAB. PMID- 7479389 TI - Raloxifene (LY156758) produces antimetastatic responses and extends survival in the PAIII rat prostatic adenocarcinoma model. AB - The benzothiophene antiestrogen, raloxifene (LY156758), has selective estrogen pharmacological antagonist activity in rats. The PAIII rat prostatic adenocarcinoma model was used to evaluate the effects of this agent on the lymphatic and pulmonary metastasis and survival in tumor-bearing male Lobund Wistar (LW) rats. Raloxifene was inactive against colony formation of PAIII cells in vitro. Similarly, following subcutaneous (s.c.) implantation of 10(6) PAIII cells in the tail, s.c. administration of raloxifene (2.0, 10.0, or 20.0 mg/kg/day) for 30 days failed to demonstrate cytoreductive activity against primary tumor growth in the tail. However, in these same animals, raloxifene administration produced significant (P < 0.05) inhibition of PAIII metastasis from the primary tumor in the tail to the gluteal and iliac lymph nodes (maximal responses = 89% and 81% from control values, respectively). PAIII metastasis to the lungs was significantly inhibited by raloxifene treatment. Numbers of pulmonary foci in PAIII-bearing rats were significantly (P < 0.05) reduced by raloxifene administration in a dose-related manner (maximal reduction = 97% from control values). In these animals, maximal regression of 20% for ventral prostate and 21% for seminal vesicle were also seen after raloxifene administration (P < 0.05 for both). Coadministration of E2B and raloxifene had no consistent antagonistic effect upon the antitumor responses produced by raloxifene. Raloxifene (40.0 mg/kg/day for 28 days) produced marked decreases in PAIII metastasis in the lymphatic and pulmonary components. Continued administration of the compound produced significant (P < 0.05) extension of survival of PAIII bearing rats. Further studies are needed to define the maximal antitumor efficacy and the mechanism of action of raloxifene in urogenital solid tumor animal models. These data support the contention that raloxifene represents a class of active antimetastatic agents with potential efficacy in the treatment of hormone insensitive human prostatic cancer. PMID- 7479391 TI - Prostate specific antigen in a community-based sample of men without prostate cancer: correlations with prostate volume, age, body mass index, and symptoms of prostatism. AB - The correlation between both prostate specific antigen levels (PSA) and prostate specific antigen density (PSAD) and age, prostate volume parameters, body mass index, and the International Prostate Symptom Score (IPSS) were studied in a community-based population. A sample of 502 men aged 55 through 74 years was evaluated, excluding those with a serum PSA above 10 ng/ml, those with biopsy proven prostate cancer, and those who had previously undergone a prostate operation. PSA and PSAD did not correlate with the body mass index. Weak correlations were found between PSA and age (r = 0.25; P < 0.001), PSAD and age (r = 0.17; P < 0.001) and between PSA and the total prostate volume (r = 0.58; P < 0.001). PSA did not correlate independently with age after adjustment for volume (P = 0.22). The finding that PSAD correlates with age (r = 0.17; P < 0.001) is partly explained by the incomplete volume adjustment of PSAD which is proved by the positive correlation between PSAD and prostate volume (r = 0.26; P < 0.001). In the main target age-range for prostate cancer screening there is a poor basis for the use of age-specific reference values or volume adjustment for PSA levels in order to increase the clinical usefulness of this serum marker. Comparison of the results of the present study and studies conducted in others regions shows that there may be significant differences in PSA values per age stratum. Further studies are needed to clarify the reasons for these differences. PMID- 7479392 TI - Work environment and prostate cancer risk. AB - A case-referent study of 345 prostate cancer cases and 1,346 referents was carried out in the Netherlands to investigate the relationship between work environment and prostate cancer risk. Cases were selected from the Cancer Registry of the Comprehensive Cancer Centre IKO. Referents (men diagnosed with benign prostate hyperplasia) were recruited with assistance of the pathology laboratories in the IKO region. Questionnaires were mailed to all subjects to obtain information on their work history and occupational exposure. Moreover, workers in farming (n = 323), and in metal work and maintenance (n = 340), were requested to complete short supplements to the questionnaire inquiring in more detail into specific types of exposure. Significantly elevated risks were found for work in food manufacturing and for bookkeepers. Significantly elevated odds ratios (OR) were also observed for jobs held between 1960 and 1970 in administration, in storage, or as farm laborer. In addition, a statistically significant excess risk was found for subjects who reported frequent occupational exposure to cadmium. Cases who worked in farming applied pesticides during significant more days per year than the referents did. A nonsignificantly elevated OR was found for maintenance of tractors and agricultural machinery. Among metal workers, mechanics, and repairmen, nonsignificantly increased ORs were observed with regard to the use of acids, solvents, iron, and steel, and for welding and maintenance of machinery. PMID- 7479393 TI - Cell proliferation in dysplasia of the prostate: analysis by PCNA immunostaining. AB - Patterns of cell proliferation in the prostate were compared between benign epithelium and dysplasia. Proliferating cell nuclear antigen (PCNA) immunostaining was used to quantitate proliferation, and basal cells were tallied separately from secretory cells with the aid of keratin immunostaining. Using a novel technique, absolute cell densities (cells/mm) were determined and used to calculate growth fractions. In benign epithelium, 83% of PCNA+ cells were basal cells, while only 7% of PCNA+ cells in dysplasia were basal cells and there was a clear separation between groups. This dramatic shift of the proliferative compartment to the secretory cells in dysplasia was accompanied only by a moderate increase in overall secretory cell density and moderate reduction in basal cell density, but these ranges overlapped those of benign epithelium. The median PCNA+ secretory cell "growth fraction" was 0.12% in benign epithelium and 1.06% in dysplasia. The findings presented give further support to the concept that dysplasia represents an evolutionary stage in the malignant transformation of prostatic epithelium. The patterns of change in PCNA immunostaining may reflect certain aspects of the biologic nature of malignant transformation. PMID- 7479394 TI - Induction of urinary plasminogen activator by retinoic acid results in increased invasiveness of human prostate cancer cells PC-3. AB - Overproduction of uPA by prostate cancer cells in vivo results in tumor invasiveness and osteoblastic skeletal metastasis due to its mitogenic actions in osteoblasts. In the present study we have examined the effect of several growth factors and steroid hormones on regulating uPA gene expression in the human prostate cancer cell line (PC-3). Treatment of these cells with dexamethasone (Dex) caused a decrease, whereas epidermal growth factor (EGF) and fetal bovine serum (FBS) increased uPA expression in a dose-dependent manner. Trans retinoic acid (RA) also induced uPA mRNA and protein production in a dose-dependent manner (10(-6) to 10(-9) M). This increase was seen as early as 2 hr of treatment until 48 hr. Dex treatment resulted in decreased tumor cells invasiveness, whereas exposure to EGF and RA caused an increase in the invasive capacity of PC-3 cells. These studies should help to better understand the control mechanism of uPA expression in prostate cancer, where uPA has been implicated as a major pathogenetic factor. PMID- 7479395 TI - Influence of dihydrotestosterone, epidermal growth factor, and basic fibroblast growth factor on the cell kinetics of the PC3, DU145, and LNCaP prostatic cancer cell lines: relationship with DNA ploidy level. AB - The cell kinetics (percentage of cells in the S+G2 phases of the cell cycle) and the DNA ploidy levels (nuclear DNA content) were determined in 108 samples each of the PC3, DU145, and LNCaP prostate cancer models. This was carried out by means of the digital cell image analysis of Feulgen-stained nuclei. Two to three hundred cell nuclei were analyzed for each of the 324 samples under study. The three cell lines were submitted to experimental conditions including the addition of dihydrotestosterone (DHT), epidermal growth factor (EGF), and basic fibroblast growth factor (bFGF), either alone or in combination, to the culture media. The results show that under the present culture conditions, the PC3 cell line was DHT , EGF- and bFGF-insensitive. In contrast to what is generally reported in the literature, the DU145 cell line was DHT- and EGF-sensitive under the present culture conditions, but bFGF-insensitive. The LNCaP cell line was DHT-sensitive, but EGF- and bFGF-insensitive. While mainly tetraploid, the three cell lines nevertheless exhibited a significant level of heterogeneity in their nuclear DNA content distributions. Indeed, the proportions of non-tetraploid (diploid, hyperdiploid, triploid, hypertriploid, hypertetraploid, polymorphic) DNA histograms were 14% in the PC3, 16% in the DU145, and 29% in the LNCaP cell lines. These results suggest that the DNA ploidy level would not influence the hormone sensitivity level in the cell lines since they had significantly distinct hormone sensitivity profiles while remaining mainly tetraploid. PMID- 7479397 TI - Examining trends. PMID- 7479396 TI - Stimulation of human prostate cancer cell lines by factors present in human osteoblast-like cells but not in bone marrow. AB - Secondary deposits of prostate tumours are frequently found in the skeleton where they produce osteoblastic lesions. In this study both osteoblast-like cells and bone marrow from the proximal femur have been cultured to determine whether or not they can release factors which could support the growth of secondary prostate tumours. Media conditioned by both osteoblast-like cells (OBCM) and bone marrow were examined for their potential to stimulate prostate carcinoma cell lines. Whilst the results obtained demonstrated that OBCM could enhance the growth of both the hormone sensitive (LNCaP) and hormone unresponsive (PC-3 and DU-145) prostate carcinoma cell lines, no proliferative effect could be shown on cell lines derived from cancers of the breast, bladder, and liver. Significantly, media conditioned by either bone marrow or human skin fibroblasts also had no effect on the growth of prostate carcinoma cell lines. This study supports the possibility that the proliferation of prostate cancer cells at secondary skeletal sites, in vivo, may be due to osteoblast derived factors. PMID- 7479398 TI - Let's play balls ... under OUR rules. PMID- 7479399 TI - Tibial pilon fractures: a comparative clinical study of management techniques and results. AB - Thirty-eight consecutive pilon fractures were reviewed retrospectively to compare the radiographic and clinical results with the original injuries based on three different treatment options: external fixation only, external fixation with limited internal fixation, and internal fixation only. Dates of injury were from February 1985 to February 1989. Treatment method was the surgeon's preference. The mean follow-up time was 28 months. Results were tabulated by clinical and radiographic scores. In general, simple fracture types had good results; complex ones did less well. In this study, fracture severity appeared to be the key variable in outcome. PMID- 7479400 TI - Closed reduction and percutaneous stabilization of tibial plateau fractures. AB - Twenty-eight patients with tibial plateau fractures treated by closed reduction using forceful traction and percutaneous stabilization with pins or screws were reviewed. For six bicondylar fractures, transfixion pins with an external fixation device were used. In six cases, an arthroscopic exam was also done. For 21 patients, external support was used postoperatively. Twenty-one patients were followed an average of 26 months. All fractures healed. Functional ratings were: 14 excellent, 6 good, and 1 poor. Radiographic ratings were: 11 excellent, 9 good, and 1 fair. For fractures with large fragments without excessive comminution or severe central depression, this technique yields good results with minimal surgical morbidity. PMID- 7479401 TI - Compartment pressure monitoring during intramedullary fixation of tibial fractures. AB - Twenty-six legs in 25 patients were monitored prospectively for compartment pressures during intramedullary nailing of open and closed tibial shaft fractures. Twenty-three patients were treated within 24 hours of admission. Twenty-three unreamed and three reamed intramedullary rods were utilized. Compartment pressures were measured initially, following fracture reduction, and during reaming. Pressures were also measured at the completion of nailing with the ankle dorsiflexed and at rest (plantar flexed) to determine positional effects on compartment pressures. Nine fractures (35%) were found to have persistently elevated pressures (> 40 mm Hg) and underwent immediate four compartment fasciotomy. The remaining group of 17 fractures (65%) was monitored throughout the intraoperative period. No patient monitored intraoperatively developed postoperative compartment syndrome. PMID- 7479402 TI - Expanding the use of the ankle distractor in the treatment of complex ankle fractures. AB - Use of the ankle distractor with ankle arthroscopy is well known to allow improved evaluation of the intraarticular surfaces. A less common application of the ankle distractor is for the treatment of complex ankle fractures. We evaluated the use of the ankle distractor in the treatment of six patients with ankle trauma. Five of these cases involved comminuted intraarticular fractures of the distal tibia, and one case involved a malunion of an ankle fracture with widening of the mortise. In each case, the ankle distractor increased the exposure of the ankle joint and allowed better access for fracture fragment reduction. There were no complications associated with the use of the distractor. The technique of using the ankle distractor in these situations is described. PMID- 7479403 TI - Adolescent femoral shaft fractures: rigid versus flexible nails. AB - Twenty-one skeletally immature patients between 11 and 16 years of age were treated operatively for a unilateral femoral shaft fracture. Eleven patients underwent fixation with flexible nails and 10 with rigid nails. The patients were studied retrospectively to determine the similarity of the groups. Each method of fixation was then compared to determine the technical ease and final outcome. Both methods gave excellent final radiographic alignment with minimal complications. Flexible nailing required much less operative time and less fluoroscopy time. Estimated cost of using Ender nails is much less than using Russell-Taylor interlocking nails. PMID- 7479404 TI - Compression plating of acute femoral shaft fractures. AB - Although intramedullary fixation has standardized the management of femoral shaft fractures, compression plating may offer a viable alternative. A retrospective review of acute femoral shaft fractures managed by open reduction and compression plating between 1980 and 1987 revealed 71 fractures in 69 patients. There were 58 closed fractures, and six grade I and seven grade II open fractures. All patients received prophylactic antibiotics. Sixty-nine percent of patients were bone grafted. Sixty-six of 71 fractures (93%) healed uneventfully. The average time until union was 16 weeks (range: 23 to 72). Complications included two nonunions, two refractures, and one broken implant (7%). Careful adherence to the principles of prophylactic antibiotics, medial cancellous bone grafting, and meticulous soft tissue technique can significantly reduce the complication rate of compression plating. PMID- 7479405 TI - Percutaneous osteosynthesis with two screws in treating femoral neck fractures. AB - In a consecutive, retrospective study, 182 patients were followed 1 year after surgical treatment of a fractured femoral neck. The patients were admitted on alternate days to the neighboring departments of orthopedic surgery O and phi, respectively. In Department O, the patients were submitted to sliding screw-plate (SSP) osteosynthesis, while patients in Department phi were treated with two percutaneously introduced screws. Our results showed that double-screw osteosynthesis can be performed significantly quicker and with less blood loss than application of SSP. Healing of the fractures in the two groups did not differ significantly, nor did the cumulative rate of failure or the frequency of secondary surgical procedures. We find that, judged from these parameters, percutaneously introduced double-screw osteosynthesis is at least as effective as SSP osteosynthesis. PMID- 7479406 TI - Lower limb salvage: microvascular reconstruction of post-traumatic soft tissue and skeletal defects. AB - This study reviews the principles of reconstructive microsurgery for lower limb salvage after severe open tibial fractures. Initial decision-making principles are reviewed with reference to useful scoring systems directed at appropriate patient selection for limb salvage. Soft tissue reconstruction is discussed with emphasis on timing and selection. Skeletal reconstruction focuses on available options and indications. PMID- 7479407 TI - Anterior stabilization of thoracolumbar burst fractures using the Kaneda device: a preliminary report. AB - Anterior decompression and fusion is a valuable technique in the treatment of thoracolumbar burst fractures. Anterior instrumentation has evolved to correct deformity and stabilize the spinal segments during decompression and bone grafting as a single-stage procedure. One anterior device developed by Kaneda has been used in our institution since 1989. This study is an initial review of our experience with this instrumentation. Anterior decompression and fusion augmented with the Kaneda device was performed in 20 patients with thoracolumbar burst fractures. Timing of surgery was early (< 15 days) in 13 patients, intermediate (16 to 120 days) in 2, and delayed (> 121 days) in 5. Sixteen patients were neurologically intact (Eismont grade D), 3 with distal weakness (Eismont grade B or C) and 1 with complete paraplegia (Eismont grade A). All patients with deficits recovered one Eismont grade after anterior decompression and fusion. Complications included pulmonary problems in 2 patients, a thoracic duct laceration requiring ligation, and a sympathectomy effect of the lower extremity. One patient had a pulmonary embolus and developed a hematoma at the graft harvest site while on anticoagulants with resultant meralgia paresthetica. There have been three screw failures and one definite pseudarthrosis. Anterior decompression and fusion supplemented by the Kaneda device was performed on 20 patients with thoracolumbar burst fractures. The average correction of kyphosis was approximately 50% acutely, with loss of approximately 50% of correction at follow up. In some patients, coronal plane deformity was created by the device. No effect on neurologic recovery could be shown with this small series. Perioperative complication rate was 30%, and 1 patient developed pseudarthrosis. PMID- 7479408 TI - A comparison study of treatment of thoracolumbar fractures using the ACE Posterior Segmental Fixator and Cotrel-Dubousset instrumentation. AB - The results of 26 consecutive thoracolumbar fractures treated with Cotrel Dubousset instrumentation (CDI) (n = 12) or the ACE Posterior Segmental Fixator (n = 14) with a mean follow up of 20.2 months were analyzed. Preoperatively, no statistically significant difference was noted between the two treatment groups. Postoperatively, no statistically significant difference was noted for improvement of kyphosis (mean: -6.00 degrees ACE, 1.92 degrees CDI), vertebral body height (mean: 17.86% ACE, 18.83% CDI), vertebral body angle (mean: -6.21 degrees ACE, -5.42 degrees CDI), or estimated blood loss (mean: 1544 cc ACE, 1620 cc CDI). All patients with incomplete paraplegia improved by at least one Frankel grade. Statistically significant differences were noted in operative time (mean: 269 minutes ACE, 357 minutes CDI, P < .0005), and in the number of instrumented levels (mean: 3 ACE, 5.8 CDI). All patients exhibited solid fusion radiographically. Thoracolumbar fractures can be effectively treated by either CDI or the ACE Fixator. The ACE Fixator has the advantage of sparing motion segments and decreased operative time. PMID- 7479409 TI - A computer tomographic classification system for the displaced intraarticular fracture of the os calcis. AB - The treatment of the displaced intraarticular fracture of the os calcis continues to be controversial. One of the reasons for this is the lack of a workable classification system which would allow comparison among different treatment modalities. Plain radiographs are unable to depict the complex three-dimensional pathology of this fracture. Computed tomography (CT) scanning, however, has the potential to quite accurately depict all components of this injury. At our hospital, a five-part, CT-based classification system has been utilized. This system suggests which fractures will do well with conservative care, and which fractures are amenable to operative stabilization. PMID- 7479410 TI - The difficult diagnosis of posterior tibialis tendon rupture in sports injuries. AB - Posterior tibialis tendon rupture is a diagnosis that is often missed in sports related injuries. This is thought to be secondary to the nonspecific clinical findings in healthy, active individuals, and the lack of any laboratory or radiographic test to reliably confirm the diagnosis. We report five cases of surgically confirmed posterior tibialis tendon rupture secondary to sports related trauma. Based on our review of these patients, the diagnosis of posterior tibialis tendon rupture should be strongly suspected in the adult patient presenting with a history of a twisting ankle injury, especially in the setting of high-impact loading and generalized medial ankle pain and swelling. A flexible asymmetric pes planus and forefoot pronation deformity with absence of posterior tibialis tendon function on manual testing is seen on examination. The patient is usually unable to perform ipsilateral single leg heel rise and has less severe pes planus of the contralateral foot. This study reviews the presentation, pathophysiology, diagnosis, and treatment of posterior tibialis tendon rupture. PMID- 7479411 TI - The effect of intercarpal joint motion on wrist motion: are there key joints? An in vitro study. AB - The contribution of each intercarpal joint individually and as a member of a regional group to total wrist motion was analyzed in 10 fresh, frozen cadaver specimens. Each intercarpal joint had an effect on the total wrist motion, but the contribution of each to the different planes of motion was not equal. The scaphoid-capitate joint had a governing effect on the motion of a region comprised of scaphoid-trapezium, scaphoid-trapezoid, and scaphoid-capitate. The lunate-capitate joint had a governing effect on the motion of a region comprised of the lunate-triquetrum, lunate-capitate, triquetrum-hamate, and hamate-capitate complex. Elimination of motion in either of the key governing joints affected total wrist motion the same as elimination of motion in all the joints in that region. Intercarpal fusion for treatment of ligament injury of the proximal intercarpal row may be limited to the scaphoid-capitate or lunate-capitate joint. There is no need to attempt fusion in all joints of either of the two regions. PMID- 7479412 TI - Cervical spinal fractures in Alpine skiers. AB - A 5-year review of skiing injuries in the Aspen/Snowmass area revealed 18 cervical fractures. This represents a yearly average of 3.6 fractures (range: 2 to 5). There were 16 men and 2 women identified, with an average age of 40.8 years (range: 20 to 73). Using estimates of skier days and total injuries, cervical fractures represented approximately 0.1% of all skiing injuries. Six of 18 injuries (33%) were avulsion injuries of the spinous processes and were treated symptomatically with a soft collar. However, five patients had neurologic involvement secondary to fracture displacement, two with lasting quadriparesis. In addition, there was one death attributed to cervical fracture. Forty-four percent of the patients had associated injuries, most commonly facial and head (33%). The three oldest patients (average age: 67 years, range: 57 to 73) had three of the more serious injuries. Two of these patients were the quadriplegics, and a third required halo stabilization of bilateral C2 facet fractures. Typically, those patients who were involved in collisions with immovable objects (trees or fences) had more severe injuries and a greater chance of neurologic involvement. PMID- 7479414 TI - Use of an infra-meniscal portal for complex posterior horn meniscal resection. PMID- 7479413 TI - Anterior glenohumeral dislocation with ipsilateral humeral shaft fracture. PMID- 7479415 TI - Bilateral dorsal midtarsal dislocations. PMID- 7479416 TI - The role of hip arthroscopy in the diagnosis and treatment of hip disease. AB - Injuries to the hip are commonly encountered by orthopedic surgeons and will, for the most part, respond well to conservative therapy. When a patient's hip pain has not responded to the appropriate treatment and radiographic measurements have not shown any specific pathology, arthroscopy of the hip can be a valuable diagnostic and therapeutic procedure. We retrospectively reviewed 94 consecutive patients who experienced refractory hip pain that persisted longer than 6 months (average: 2.1 years) and who had an average follow up of 2.5 years. Statistical analysis was performed using Fisher's Exact Test (P value) and Pearson (r value) correlation coefficient to compare preoperative symptomatology and physical examination findings with intraoperative pathology. Statistically significant results were obtained when comparing preoperative symptoms of a painful hip click and mechanical symptoms of locking with acetabular labral injuries (P = .000, r = .896) and loose bodies (P = .000, r = .896), respectively. Radiologic studies were nondiagnostic in 76% of patients in this series. Hip arthroscopy can be a valuable and viable procedure in the diagnosis and treatment of protracted or progressive hip pain lasting more than 6 months. PMID- 7479417 TI - Radiologic case study. Glomus tumor. PMID- 7479419 TI - Thromboembolic complications following trauma surgery: incidence and outcomes. AB - Thromboembolic disease is an unusual condition in the general community. The incidence of symptomatic acute deep vein thrombosis (DVT) is very low in the general population before the age of 50 years. After the age of 50 years, the annual incidence increases. Thromboembolic disease is, however, a major cause of death and morbidity among hospitalized patients. In this setting, it has been estimated that pulmonary embolism (PE) kills about 100,000 Americans each year, and contributes to the death of another 100,000. It is probably the most common preventable cause of death in hospitalized patients. Thromboembolic disease is a cluster of related conditions, many of them clinically silent. DVT and PE manifest themselves with very general symptoms, making the clinical diagnosis (symptoms and physical signs) insensitive and nonspecific. PMID- 7479418 TI - Preventing DVT following total knee replacement: a review of recent clinical trials. AB - Deep vein thrombosis (DVT) following total hip or knee arthroplasty is a frequent complication which can cause fatal pulmonary embolism (PE). Although many different approaches to its prevention have been attempted, low-dose warfarin has become the standard treatment for the prevention of thromboembolic disease following total hip and knee arthroplasty. Although subcutaneous heparin can reduce the incidence of thromboembolic disease, it has been associated with excessive bleeding complications. The introduction of low molecular weight heparins promises fewer bleeding complications with efficacy equal to that of warfarin. We wish to report the results of a randomized, prospective study comparing a low molecular weight heparin with low dose warfarin in the prevention of thromboembolic disease following total knee arthroplasty (TKA). PMID- 7479421 TI - Treatment of established deep vein thrombosis: a review of the therapeutic armamentarium. AB - Although it has never been evaluated by an appropriately designed clinical trial, there is a strong impression that acute symptoms of deep vein thrombosis (DVT) and pulmonary embolism (PE) are improved by heparin treatment. The first experimental evidence showing that anticoagulants were effective in reducing mortality from PE came in landmark study by Barritt and Jordon in Lancet in 1960. In 1986, Hull et al demonstrated that failure to attain an adequate anticoagulant effect with heparin in patients with proximal vein thrombosis is associated with a high rate of recurrence. Current practice favors admission to the hospital of patients with venous thromboembolism to be treated with intravenous (IV) heparin for at least 4-5 days, overlapped with initiation of warfarin. Four randomized trials evaluating continuous intravenous (CIV) vs. subcutaneous (SQ) heparin administration in the treatment of DVT found no significant differences in the observed rates of clinically important events. Thus, SQ route heparin is as efficacious and would allow treatment at home. Low molecular weight heparins, when released by the FDA for this indication, will make this more feasible. PMID- 7479420 TI - Deep vein thrombosis prophylaxis in the outpatient setting: preventing complications following hospital discharge. AB - Venous thromboembolism is an important complication in patients undergoing major orthopedic surgery. Without prophylaxis, there is a 50% to 60% incidence of deep vein thrombosis (DVT), a 10% to 30% incidence of proximal vein thrombosis, and a 1% to 2% rate of fatal pulmonary embolism after total hip replacement. The incidence of DVT after knee arthroplasty is even higher, ranging from 55% to 70% and, although there are no accurate figures on the incidence of fatal pulmonary embolism, it is likely to be similar to that following hip arthroplasty. Low molecular weight heparins (LMWHs) have been shown to be effective and safe in the prevention of venous thromboembolism in patients undergoing major orthopedic procedures. In comparative studies, LMWHs are more effective than adjusted dose heparin or warfarin without increased risk of bleeding. PMID- 7479423 TI - Critical care pathways in thromboembolic disease. AB - Critical Care Pathways are outlines of each day's expected interventions and treatments for a particular process. They are part of a larger concept of case management, as an extramural, multidisciplinary approach to a patient's problems, with details suitable to their personality and case. This constitutes the overall concept of managed care. Rather than DVT, this presentation focuses on thromboembolism. I think you'll see why I chose to emphasize this complication instead. Modern medicine has been turned upside down; the rules are different, expectations are different. I would suggest to physicians who are resisting the CCP concept that it may be your salvation as much as it is a paperwork torment. It's clearly a way to provide better care for your patients. PMID- 7479422 TI - Consortium data: comparative efficacy of low molecular weight heparin and warfarin following total hip replacement. AB - The Consortium Study is a prospective, randomized research project designed to extend the medical community's understanding of the incidence of thromboembolic disease following hip replacement surgery. The study's methodology of assessing deep vein thrombosis (DVT) and pulmonary embolism (PE) is by clinical symptomatology. Once clinical symptoms occur, diagnostic confirmation includes noninvasive techniques such as duplex ultrasonography, venography, VQ lung scan, and/or lung angiography. The study is designed to recruit a cohort of 4500 patients and will involve 150 medical centers in the United States. The final conclusion and definitive data of the incidence of DVT and PE, when comparing enoxaparin with warfarin, will be available after the full complement of 4500 patients is recruited. This study will demonstrate the incidence of thromboembolic disease during the acute hospital phase and the medium-term phase for the patient undergoing elective hip replacement. PMID- 7479424 TI - Investigations on preparation and properties of modified polyacrylamide hydrogels for application as wound dressing materials. AB - Investigations on the influence of the polymerization conditions and feed ratio as well as various modification of the composite polyacrylamide-agar hydrogels on their properties has been presented. Results of the model studies have been utilized in the optimalization of the manufacturing parameters and procedures of the preparation of hydrogel dressing foils and granulated dressing gels. PMID- 7479425 TI - [Alloplastic hydroxyapatite implants in treatment of periodontitis]. AB - The authors estimated the porous hydroxyapatite alloplasts in the treatment of periodontitis in a clinical and radiological way. The material has been used in 30 patients who had bone pockets deeper than 6 mm. Together, 46 bone pockets were filled. We obtained reduction of the depths of the pockets of 3.02 mm, decrease of the degree of the teeth mobility and the improvement of Engelberger, Marthaler and Raitetschak's radiological indicator by 13%. The results were compared with other works describing the usage of this material in stomatology. Basing on the literature the authors also presented the histological changes in periodontium tissues after the usage of tricalcium phosphate and compact and porous alloplastic hydroxyapatite ceramics. PMID- 7479426 TI - Does the liquid method of electret forming influence the adhesion of blood platelets? AB - This work presents the results of the effect of the electric charge on the adhesion of blood platelets. All experiments were carried out on polyethylene foil. The liquid method was used to form electrets. The evaluation of the electret effect influence on the adhesion of blood platelets was made on the basis of the observation of the electret surface after the contact with fresh citrate human blood group O Rh+ in an electron scanning microscope. Experimental results confirmed the essential influence of the electric charge on the process of adhesion of blood platelets. It was noticed that the preliminary aging of electrets decreases the density of the surface charge and improves the athrombogenic characteristics of polyethylene foil. PMID- 7479427 TI - [Analysis of the effects of selected polymers on morpho-dynamic features of rat hepatocytes]. AB - The thesis shows the results of the research on the influence of the film-form polymers on the hepatocytes in rats. The polymers were composed of polyethylene (PE), polyvinyl chlorides (P-25, P-47), polyesters (A, B, E) and irradiated L7 collagen. The aim of the research was to establish whether the changes in the specific morpho-dynamic characteristics-which prove the changes in the hepatocyte metabolic activity in rats-could be used to assess the usability of the above mentioned polymers for medicine. The evaluation of the research findings was analysed on the basic of the stereometric methods and factor analysis. On the basic of the obtained results, polyvinyl chlorides P-25 and P-47 have been evaluated most favourably as far as their possible application in medicine is concerned, whereas polyester E shows the most negative characteristics for such an application. PMID- 7479428 TI - [Use of irradiation technique for obtaining and modifying biopolymers. Review]. AB - A review of papers concerning application of radiation techniques to the biopolymers production is presented. The nature of electron and gamma irradiation influence on polymers is outlined. Advantages of the method from the point of view of biocompatibility and bio-functionality of biopolymers are underlined. Among them the most important are the following: chemical purity of products, high efficiency of the method, expanded influence on polymers' structure, usefulness in the graft copolymerization, ability of avoiding enhanced temperature during polymerisation and sterility of products. Examples of biopolymers obtained or modified by means of irradiation techniques are gathered. PMID- 7479429 TI - Amphotericin B. PMID- 7479430 TI - Melanomas in children. AB - Although rare, melanomas do occur in children and adolescents. Pediatricians should be aware of the clinical features of melanoma and the risk factors for developing this malignancy. Children at high risk for melanoma should have at least annual cutaneous examinations in search of suspicious lesions. If a lesion is suspected of being a melanoma, it should be removed surgically and submitted for pathologic examination. Education of parents and children about the deleterious effects of ultraviolet light affords a means of counteracting the increasing incidence of melanoma at the grassroots level. The use of sunscreens, hats, and other protective clothing and the judicious timing of daily solar exposure should serve to prevent sunburning, limit tanning, and decrease the incidence of melanoma and other more common cutaneous malignancies, such as basal cell carcinoma and squamous cell carcinoma. PMID- 7479431 TI - Eating disorders among children and adolescents. AB - Anorexia nervosa and bulimia nervosa represent extreme manifestations of weight control patterns in adolescents. When they occur in children, they often are associated with more severe psychiatric or family dysfunction. However, rather than framing these conditions as purely psychiatric problems, it is more useful to approach them in terms of development. Then, the pediatrician can be positioned to address the various biologic, psychological, and social issues presenting in the individual. Furthermore, the pediatrician already is familiar with the principles of managing chronic illnesses, and eating disorders frequently require months to years of treatment. Finally, for each patient in a pediatrician's practice who has an eating disorder, several other patients will develop unhealthy habits with respect to eating, food choices, and weight control. By being familiar with the larger issues associated with weight loss, rather than the more narrow topic of eating disorders, the pediatrician will be able to help all patients remain healthy in a culture of thinness, regardless of their diagnosis. PMID- 7479432 TI - Osteomyelitis. PMID- 7479433 TI - Status epilepticus. PMID- 7479434 TI - Principles of inborn errors of metabolism: an exercise. AB - Many inborn errors of intermediary metabolism present during periods of poor oral intake or intercurrent illness. Under these conditions, the body switches to a predominantly catabolic state, when glycogen, fat, and protein are mobilized as alternative energy sources. If a genetic defect already exists in any of these catabolic pathways, the problem is accentuated because the catabolic state demands a high flux of metabolite through these pathways. Therefore, a cardinal principle in the management of children who have IEM is to prevent them from lapsing into a catabolic state. This can be done through: 1) aggressive nutrition management to promote positive nitrogen balance and decrease accumulation of toxic metabolites, and 2) immunizations to minimize the possibility of viral or bacterial illness. When catabolism does occur, high-dose glucose infusion is a potent method for turning off the catabolic state. This simplistic view of the catabolic pathways neglects many details in each of the metabolic pathways and diseases cited in the article and questions. Nevertheless, it should provide a useful framework for explaining some of the clinical findings encountered in caring for this special group of patients. A partial list of more commonly encountered IEMs is contained in the Table. The different parts of the biochemical pathways discussed previously are combined into Figure 6. Hopefully, for some readers, the Figure also will lead to the affirmation that biochemistry is not so bad after all! PMID- 7479435 TI - Poverty and health. PMID- 7479436 TI - Antibiotic drug levels. PMID- 7479438 TI - [Looking for a needle in a haystack. Identification of differentially expressed genes]. PMID- 7479437 TI - [Structure and properties of transcription IIIA from Xenopus laevis]. PMID- 7479439 TI - [Regulation of fibrinogen molecule synthesis]. PMID- 7479440 TI - [Mosaic proteins]. PMID- 7479441 TI - [Flippases--phospholipid transporting proteins]. PMID- 7479442 TI - [Glycosaminoglycans--structure and function]. PMID- 7479443 TI - [Lipids--liposomes--biological membranes]. PMID- 7479444 TI - [Regulation of the cell cycle--involvement of protein inhibitors of cyclin dependent kinases]. PMID- 7479445 TI - [Regulation of Escherichia coli heat shock response]. PMID- 7479446 TI - Functional colonic and anorectal disorders. Detecting and overcoming causes of constipation and fecal incontinence. AB - Evaluation of constipation and fecal incontinence begins with comprehensive history taking, which may include overcoming a significant psychosocial barrier. Before functional anorectal diseases can be controlled, the fundamental cause must be determined and any underlying disorder corrected. In constipation, a colonic or anorectal motility disorder is often the cause; about half of refractory cases are the result of obstructive defecation. In fecal incontinence, dysfunction of several anatomic or physiologic mechanisms may be the cause. Anorectal manometry is useful in assessment in both disorders. Other helpful tests are colonic-transit measurement in constipation and electrophysiologic tests and defecography in fecal incontinence. Treatment of constipation often includes dietary measures and use of laxatives or prokinetic agents; fecal incontinence may respond to bulking or antidiarrheal agents. In both disorders, some patients have responded to the recently described technique of neuromuscular conditioning with biofeedback. In some cases, surgical repair must be considered. PMID- 7479448 TI - Confidential help for impaired physicians. PMID- 7479447 TI - Congestive heart failure. Considerations for primary care physicians. AB - Initial evaluation of congestive heart failure (CHF) should emphasize a search for curable and precipitating causes. Echocardiography is mandatory if CHF is a serious consideration after initial evaluation. Digitalis, diuretics, and vasodilators in the form of angiotensin-converting enzyme (ACE) inhibitors are the mainstays of pharmacologic therapy. Early use of ACE inhibitors for asymptomatic left ventricular dysfunction and established CHF lessens morbidity and mortality. The number of expensive hospitalizations can be reduced by patient education and institution of a flexible therapeutic program guided by the patient's symptoms and daily weigh-ins. PMID- 7479449 TI - Health risks in young athletes. PMID- 7479450 TI - Vaccination update. Hib, hepatitis, polio, varicella, influenza, pneumococcal and meningococcal disease. AB - Immunization of infants and children is the most effective strategy for decreasing the incidence of some infectious diseases. Most invasive disease due to Haemophilus influenzae type b occurs before age 5 years, and routine vaccination of infants for hepatitis B is currently recommended because selective immunization of high-risk persons has not been feasible. Decades of use of poliovirus vaccine has effectively eliminated cases of wild-virus infection, although some vaccine-related cases still occur. The newly approved varicella vaccine appears to be a cost-effective way to decrease infection rates in children. Improved immunization rates for influenza and pneumococcal and meningococcal diseases could help decrease excess mortality in elderly persons and those with chronic illness. PMID- 7479451 TI - Recurrent corneal erosions. Management by the primary care physician. AB - The primary care physician should be aware of recurrent corneal erosion, a condition that is quite common but is often misdiagnosed. Most cases are a result of minor corneal abrasion, after which the corneal epithelium undergoes a cycle of breakdown and healing that may last for months. Use of a bland antibiotic ointment twice a day for 4 to 8 weeks may be successful, but if the patient is very uncomfortable or continues to have symptoms, referral to an ophthalmologist is appropriate. PMID- 7479452 TI - Chronic schizophrenia. Options for pharmacologic management. PMID- 7479453 TI - Pitfalls in HIV testing. Application and limitations of current tests. AB - Enzyme-linked immunosorbent assay (ELISA) and Western blot assay are the most commonly used laboratory tests for HIV infection. Both detect antibodies to HIV. ELISA results are based on detection of antigen-antibody complexes by using antibodies labeled with an enzyme that produces a color change in the presence of a specific substrate. Currently licensed ELISA tests have greater than 98% sensitivity and specificity for HIV. Western blot analysis detects antibodies to specific HIV antigens and is best used as a confirmatory test. In spite of the high sensitivity and specificity of both tests, false-positive and false-negative results do occur. Physicians should be aware of specific causes of inaccurate results. In individual cases, knowledge of the patient's history and the criteria used by the laboratory performing the test is important. PMID- 7479454 TI - Fetal alcohol syndrome. How you can help prevent it. AB - Fetal alcohol syndrome results from maternal alcohol consumption during pregnancy. Clinical manifestations include growth deficiencies, facial abnormalities, neurologic disorders, cardiac septal defects, and minor joint and limb abnormalities. Diagnosis is based on clinical findings and a history of maternal alcohol consumption of more than two drinks per day. Total abstinence from alcohol during pregnancy, especially for moderate and heavy drinkers, should be advocated before conception or as early as possible in the prenatal period. With proper education and counseling, the incidence of this unfortunate but preventable syndrome could be significantly decreased. PMID- 7479455 TI - Arteriosclerosis obliterans. Diagnostic and nonoperative treatment options. PMID- 7479457 TI - Medical education: an unwelcome guest at the healthcare table. PMID- 7479456 TI - Fingertip and nail bed injuries. Repair techniques for optimum outcome. AB - Most fingertip injuries can be treated in a procedure room, provided proper lighting and equipment are available. Sound judgment and knowledge of fingertip anatomy are essential. Determining the mechanism of injury is important, because it may indicate the degree of contamination, amount of tissue loss, and best treatment. Superficial wounds may be allowed to granulate and contract spontaneously. In children, even amputation may heal by secondary intention, with the fingertip sutured back in place as a biologic dressing. Split- and full thickness skin grafts may be appropriate, but diminished sensibility limits their usefulness on volar surfaces. Local skin flaps are indicated when the wound bed is unsuitable for grafting or when skin is needed to cover exposed bone or tendon. Direct closure may be used in amputations of 2 to 3 mm. When the nail bed is lacerated, the nail plate must be removed and the wound repaired. Any free segments of nail bed should be sutured in place as a free graft. In children, treatment should be conservative, with emphasis on preservation of digital length. PMID- 7479458 TI - Glad to be alive. PMID- 7479459 TI - Inflammatory diseases of the colon. Narrowing a wide field of symptoms and possible causes. AB - Symptoms and physical findings may indicate the severity of inflammatory disease of the colon, but detailed history taking is needed to limit the wide spectrum of possible causes. Infectious causes should be ruled out before other disease is assumed to be present. No single test is sufficient to diagnose ulcerative colitis or Crohn's disease. Laboratory testing, histologic assessment, endoscopy, radiology, and bowel studies are often necessary in differential diagnosis. Because of the systemic nature of colitis, manifestations in the musculoskeletal, ocular, dermatologic, hepatobiliary, and other systems may occur and provide clues. Treatment depends on the type and severity of disease. Sulfasalazine (Azulfidine), sulfa-free 5-aminosalicylic acid compounds, and corticosteroids are mainstays of treatment of ulcerative colitis and Crohn's disease. Supportive care and judicious use of antimicrobial therapy are usually effective in colitis due to bacterial, parasitic, and sexually transmitted infections and are useful for symptoms caused by colonic ischemia and vasculitis. Colitis resulting from radiation therapy may present several years after the procedure and can be difficult to diagnose and treat. In many cases of inflammatory colon disease, especially chronic conditions, consultation with a gastroenterologist is highly recommended. PMID- 7479460 TI - Anorectal disorders. Five common causes of pain, itching, and bleeding. AB - Hemorrhoids are a common source of anorectal symptoms, which are determined by their size. A helpful measure in small hemorrhoids is a high-fiber diet; in moderate lesions, ligature therapy is effective. Surgery should be reserved for large hemorrhoids. Anal fissures, probably caused by trauma to the anal canal during defecation, may persist because of a cycle of hard stools, pain, and reflex spasm. A high-fiber diet and sitz baths relieve acute fissures. Lateral partial internal sphincterotomy is usually effective when they become chronic. Perianal abscess is often caused by acute infection of the anal glands; fistula is the result of chronic infection. Swelling and induration may be present. Pain is throbbing and continuous, and perianal examination may require use of an anesthetic. Incision and drainage with follow-up to ensure resolution of infection is required. Pruritus ani may result from several contributing conditions or may be idiopathic. Restoration of dry, intact perianal skin is the treatment goal. Patients should be taught gentle hygiene and drying methods and advised to avoid caffeine or other dietary items that seem to exacerbate symptoms. Condylomata acuminata cause bleeding and pain if allowed to progress. Biopsy should be considered in patients at risk of dysplasia. Repeated application of caustic topical agents may help small lesions. Large, extensive, and persistent lesions require surgical ablation. PMID- 7479461 TI - Colorectal neoplasms. Screening can save lives. AB - Colorectal cancer is a very important cause of death and disability throughout the world. In the future, it may be possible to reduce the risk of this disease through such measures as dietary modification or use of aspirin or other nonsteroidal anti-inflammatory drugs, but this remains to be proved. It may also be possible to detect persons at high risk or those with early lesions by using noninvasive studies, such as genetic tests or detection of trace markers in the stool, but these approaches either are not yet available or are not proved in clinical trials. However, through careful and systematic clinical history taking and appropriate screening studies, neoplasms can be detected early or in the precancerous stage. Mortality from colon cancer could be reduced dramatically by applying currently available methods as part of a health maintenance program. Fecal occult blood testing and flexible sigmoidoscopy must be integrated into practice routines in the same way that Pap smears and mammography have been accepted by physicians and patients as a way to save lives. PMID- 7479463 TI - The older driver--a review. AB - There are differences between elderly drivers and younger drivers in that the elderly are more likely to have cognitive, motor and sensoriperceptual deficits affecting their driving performance. The elderly driver is more likely to have a chronic illness and to be on medication, that might adversely affect driving. The elderly driver requires more study and help either by retraining or provision of adaptations to vehicles. Fitness to drive is of clinical importance to doctors in most specialities and recent studies would suggest that doctors need to be more aware of the current guidelines on driving and be prepared to offer advice. PMID- 7479464 TI - Telling parents their child has severe congenital anomalies. AB - A large number of potential malformations exist, but the key skills in communicating with the parents of a malformed baby are the same. Dissatisfaction is common in this situation and has prompted a national initiative in improving standards of disclosure of disability and malformation--the Right from the start strategy (see appendix). Key skills include keeping the family together, ie, not removing the baby unnecessarily, knowing how parents like to be informed, listening, checking that parents have understood, being nonderogatory and, most importantly, emphasizing the positive side of their newborn child. PMID- 7479465 TI - The management of acute myocardial infarction. AB - Greater understanding of the underlying pathophysiology of acute myocardial infarction (AMI) has led to more aggressive management and lower mortality, both in-hospital and long term. AMI results mainly from thrombotic occlusion of the infarct-related coronary artery. The ensuing necrosis evolves over a 6-12 h period providing a time window for interventions designed to reduce eventual infarct size. The most appropriate interventions are those which restore coronary artery patency and hence myocardial blood flow as soon as possible. Occasionally, disruption of the occluding thrombus and compression of the underlying atheromatous lesion is best achieved by direct percutaneous transluminal coronary angioplasty. For the vast majority however, revascularisation by drug therapy is more appropriate. As soon as possible, all patients without contraindications should be offered oral aspirin and intravenous thrombolysis, usually with streptokinase but occasionally with tissue plasminogen activator. Patients in whom these agents are contraindicated should be considered for intravenous beta blockade using atenolol or metoprolol to reduce myocardial demand and hence infarct size. Patients with large infarcts, impaired ventricular function, left ventricular failure or hypertension should be considered for early angiotensin converting enzyme inhibitor therapy. Other agents may be valuable symptomatically, but have no proven role in reducing infarct size or mortality. After the first 24 h, the main aims of management are to assess the likelihood of later ischaemic events or death (risk stratification) and hence to choose appropriate long term secondary prophylaxis. PMID- 7479462 TI - Renal dysfunction associated with liver transplantation. AB - It has been known for some time that a variety of liver diseases affect kidney function, but renal dysfunction associated with orthotopic liver transplantation has received scant attention. Although the mechanisms mediating these abnormalities are incompletely defined, advances in the understanding of renal pathophysiology after liver transplantation have made it possible to develop new treatment strategies. Aggressive and early intervention to diagnose and treat renal complications associated with liver transplantation should be the goal for transplant centres. PMID- 7479466 TI - The clinical and laboratory correlates of an increased urinary 5 hydroxyindoleacetic acid. AB - Over a five-and-a-half-year period, there were 298 laboratory requests for urinary 5-hydroxyindoleacetic acid (5-HIAA). The clinical and laboratory associations of the 24 patients in which there were 43 urinary 5-HIAA 24-h collection results greater than the laboratory upper reference limit are detailed. Four were confirmed carcinoid tumours and two were phaeochromocytomas. Flushing was a prominent symptom in 46% and diarrhoea or altered bowel habit in 37%. Associated with the raised urinary 5-HIAA values were increased levels of 4 hydroxy-3-methoxymandelic acid and homovanillic acid in 14.3% and 21%, respectively, of those collections where the metabolites were requested. Diagnostic imaging was performed in 57%. While the specificity was 88%, 5-HIAA is relatively insensitive in the diagnosis of carcinoid tumours and a more widespread use of diagnostic imaging including isotope scanning with labelled metaiodo-benzylguanidine, vasoactive intestinal peptide and octreotide is suggested. PMID- 7479468 TI - Hyperkalaemia in diabetes: prevalence and associations. AB - Hyperkalaemia is associated with diabetes, but there are no recent reports of its prevalence and associations. Serum potassium concentrations were measured in all 1764 patients attending a diabetic clinic over a 12-month period and found to be > 5.0 mmol/l in 270 (15%), and > 5.4 mmol/l in 67 (4%). There was no other evident cause of hyperkalaemia in 41 of these 67 patients. These data serve to highlight the risk of dangerous hyperkalaemia in diabetic patients, particularly with concurrent administration of angiotensin-converting-enzyme inhibitors and potassium-sparing diuretics. PMID- 7479467 TI - Doubly disabled: diabetes in combination with an eating disorder. AB - We present a series of patients with both an eating disorder and diabetes mellitus and compare these to a group of non-diabetic patients from the same clinic. Significantly more of the diabetic patients had previous attempts at treatment for their eating disorder. A high incidence of diabetic complications was noted with clear implications, both clinically and economically, for early intervention. The incidence of childhood trauma was lower in the diabetic than the non-diabetic group. In the majority of patients, diabetes developed before the eating disorder, suggesting that diabetes itself may provide the vulnerability and increase the risk of developing an eating disorder. Early intervention in diabetic clinics may prevent the development of serious eating disorders. PMID- 7479470 TI - Normolipemic xanthomatosis. AB - A 20-year-old woman presented with tendon and tuberous xanthomas. Plasma lipid levels were normal. Xanthomatosis with normal lipid levels is rare. PMID- 7479471 TI - Pure white cell aplasia and health food products. AB - The safety of 'health' foods and complementary medicine products is increasingly questioned. We report a case of pure white cell aplasia developing in a patient who took a variety of such products leading to an excessive intake of zinc. Recovery was complete following the withdrawal of her 'medications'. The aetiology of the pure white cell aplasia is discussed with respect to the ingredients of the products ingested. PMID- 7479469 TI - Rapidly progressive glomerulonephritis complicating acute rheumatic fever. AB - A case of acute rheumatic fever and glomerulonephritis following streptococcal throat infection is presented. The coincidence of rheumatic fever and post streptococcal glomerulonephritis is uncommon, but well recognised. This case is of additional interest since the nephritis was crescentic. PMID- 7479472 TI - Vasospasm causing a cold hemiplegic arm. PMID- 7479473 TI - Renal calculi in primary hyperaldosteronism. AB - Increased urinary calcium (Ca++) excretion and the presence of negative Ca++ balance is well documented in primary hyperaldosteronism. However, renal calculi as a major manifestation of this disorder has not previously been described. This report describes a patient who presented with renal calculi in association with primary hyperaldosteronism. We believe that primary hyperaldosteronism was a major pathogenetic factor in the formation of renal calculi since the increased urinary excretion of Ca++ and uric acid noted at onset declined following a short term spironolactone administration and remission from renal calculi has persisted following initial nephrolithotomy and continued spironolactone therapy, which also corrected hypertension and hypokalemia, a hallmark of this disorder. PMID- 7479474 TI - Giant cell carcinoma of the lung. AB - A 50-year-old non-smoking, hypertensive female, presenting with superior vena caval compression, was found to have giant cell carcinoma of the lung. She received intensive combination chemotherapy. However she died in the following 36 hours, as a consequence of refractory hypotension. PMID- 7479475 TI - Tuberculosis of the pancreas. AB - Tuberculosis of the pancreas is extremely rare. We report a case which demonstrates the diagnostic confusion which may arise in this condition. A 55 year-old alcoholic caucasian man presented with loss of weight and obstructive jaundice. Ultrasonography, endoscopic retrograde cholangiopancreatography and computed tomography showed a mass lesion in the head of the pancreas, diagnosed as probably malignant. He underwent a Whipple's pancreatoduodencetomy and made a good recovery. Histological examination showed typical features of tuberculosis. In the absence of other foci of active disease chemotherapy was not given. He remains well 12 months after operation. PMID- 7479476 TI - Bilateral adrenal masses. PMID- 7479477 TI - Sternal mass and fever. PMID- 7479478 TI - Acute pulmonary oedema in late pregnancy. PMID- 7479479 TI - Hyponatraemic rhabdomyolysis in Addison's disease. PMID- 7479480 TI - Clinical guidelines. PMID- 7479481 TI - Are blood tests of value in the primary assessment and resuscitation of patients in the A&E department? PMID- 7479482 TI - Comparative yolk cholesterol content in four Spanish breeds of hens, an F2 cross, and a White Leghorn population. AB - Yolk cholesterol content (milligrams per gram of yolk and milligrams per egg) was studied in eggs of four Spanish breeds of hens (Castellana, Buff Prat, Vasca, and Villafranquina), an F2 from a cross between Castellana and Buff Prat (C x BP-F2), and a White Leghorn population. Birds were tested at 30 wk of age. There were differences among breeds (P < .001), cholesterol concentration being significantly lower in the C x BP-F2 (13.14 +/- .26 mg/g yolk) than in the other breeds. Villafranquina and Buff Prat showed significantly lower cholesterol (14.54 +/- .26 and 14.56 +/- .26 mg/g yolk, respectively) than the White Leghorn (16.30 +/- .26 mg/g yolk). Eggs from the Vasca breed contained significantly more cholesterol (19.09 +/- .26 mg/g yolk) than did eggs laid by the other breeds, whereas no differences were found between Leghorn and Castellana. A negative heterosis percentage of -13% was found in the C x BP-F2 when compared with the parental breed means. The reduction in cholesterol concentration per egg observed in the C x BP-F2 (220.49 mg per egg) was not related to breed differences in egg weight or to the proportion of yolk per egg. Eggs from the Vasca breed, with the lightest yolk and smallest yolk: albumen ratio (P < .05), contained the highest amount of cholesterol (304.29 mg per egg). Eggs from the White Leghorn containing the second highest amount of cholesterol (275.63 mg per egg). Thus, the variation in cholesterol content per egg was not attributable to differences in the proportion of yolk. PMID- 7479483 TI - Survey of major histocompatibility complex class II haplotypes in four turkey lines using restriction fragment length polymorphism analysis with nonradioactive DNA detection. AB - Four turkey lines were typed for MHC Class II haplotypes with restriction fragment length polymorphism (RFLP) analysis using a nonradioactive probe made from a chicken genomic clone of MHC Class II genes. The RFLP analysis detected 18 new patterns in the populations. There were three new haplotypes that had a frequency of about 10% or more in a population, whereas the rest appeared only once. The haplotype frequencies were significantly different in the E line, selected only for increased egg production, and the F line, selected only for increased body weight, compared with their respective randombred control lines. The shift of haplotype frequencies in the two selected lines seemed to be in opposite directions. One, but not the same, haplotype predominated in the selected lines, with about 50% of total haplotypes. Fewer haplotypes were frequent in the selected lines, whereas the frequencies in the control lines were relatively widely distributed, with the most frequent haplotype being below 35%. The frequency of homozygotes of the Class II haplotypes was the highest in the F line. PMID- 7479484 TI - The stability of three randombred control lines of turkeys. AB - A survey of growth and reproduction traits over eight and seven generations, respectively, is presented for three randombred control (RBC) lines of turkeys. Of the 26 traits measured, regression coefficients for the three RBC lines differed for only 8-wk body weight of females and egg weight, although the difference was in magnitude and not direction of the slopes. A common environmental trend was suggested for 8-wk male and female body weight and for 16 wk shank width of males. There was little evidence of genetic drift in these three RBC lines of turkeys. PMID- 7479485 TI - Effects of strain and rearing dietary regimens on brown-egg pullet growth and strain, rearing dietary regimens, density, and feeder space effects on subsequent laying performance. AB - The objective of this experiment was to compare the growth and development of two brown-egg strains when grown on three different dietary regimens, i.e., a "normal" step-down protein (SDP) regimen, a step-up protein regimen with a low energy starter (SUPLES), and a step-up protein regimen with a high energy starter (SUPHES). The SUPLES and SUPHES feeding regimens resulted in significantly lower BW and feed conversion, shorter sternum length, and lower tibial breaking strength than the SDP regimen. The weights of the liver, spleen, and fat pad were not different among the three regimens. The high energy starter in the SUPHES regimen significantly reduced FC below that obtained with the SUPLES program. The SUPHES regimen significantly reduced total feed cost compared with the other two feeding regimens. Both step-up protein regimens reduced total protein, energy, Ca, P, lysine, and sulfur amino acid consumption. Because tibial breaking strength was also reduced by the step-up protein regimens, the data suggest that the levels of dietary Ca and P should be increased with these programs to compensate for the reduced consumption associated with them. PMID- 7479486 TI - Treatment of commercial broiler chickens with a characterized culture of cecal bacteria to reduce salmonellae colonization. AB - The effect of treatment with a newly developed characterized continuous-flow (CCF) culture composed of 29 strains of cecal bacteria on salmonellae colonization was evaluated in commercially reared broiler chickens. Newly hatched chicks in three flocks were sprayed with CCF culture as they were placed in rearing houses and compared with untreated flocks on the same farm. Hatchery transport tray liners and rearing house feed, water, and little samples were cultured for the presence of salmonellae. Cecal samples were cultured after 3- and 6-wk growout. Skin-feather samples were cultured at 6-wk growout. Fifty-five percent of the transport tray liners were contaminated with salmonellae on the day of chick placement. At 3 wk, salmonellae serotypes present on the tray liners were widely distributed in the litter of the rearing houses. The results indicated that exposure to salmonellae occurred before culture treatment and continued in the rearing houses during the 6-wk growout period. Salmonellae cecal colonization was decreased (P < .05) in two of the treated flocks at 3 wk and diminished (P < .07) in the third treated flock compared with control flocks. At 6 wk, skin-feather contamination and cecal colonization were decreased (P < .05) in one of the treated flocks whereas no treatment effect occurred in two of the treated flocks compared with controls. The results clearly indicate the necessity of implementing integrated programs to control salmonellae in both the hatchery and rearing house environments. The CCF culture served to enhance salmonellae colonization resistance and may serve as a useful component of an integrated control program. PMID- 7479487 TI - Synergistic action of two sources of avian growth factors on proliferative differentiation of chick embryonic hematopoietic cells. AB - During embryonic development, the components of the avian immune system undergo ontogeny in several distinct organs, including the bone marrow, spleen, thymus, and bursa of Fabricius. This process is regulated and controlled by the complex interactions of various cytokines and colony-stimulating factors (CSF). The objective was to examine the action of two different sources of hematopoietic growth factors, spleen-conditioned media (SCM) and chick embryo extract (CEE), on the proliferation of hematopoietic cells from various organs and on the differentiation of progenitor cells in semi-solid culture. Spleen and bone marrow cells obtained at Day 16 of incubation responded in a dose-dependent manner to the addition of SCM and CEE alone or in combination. No proliferative effect of SCM was observed on cells obtained from embryonic thymus or bursa. Clonal analysis of bone marrow and spleen cells suggested that CEE may contain the avian equivalents of stem cell factor, interleukin-3, granulocyte-macrophage CSF, granulocyte-CSF, and macrophage-CSF. Clonal analysis of SCM cultures suggested that in addition to myelomonocytic growth factor, which affects primarily macrophage-granulocyte lineages, a thrombocyte-CSF-like activity was also apparent. The SCM alone tended to act upon committed late progenitors. The combination of CEE and SCM amplified the size and the total number of colonies obtained and appeared to act synergistically upon progenitors with a high level of proliferative potential. This response on young progenitors was confirmed when cells were cultured in CEE and SCM prior to clonal analysis. These results document the presence of thrombocyte CSF in SCM and the effect of both CEE and SCM on the proliferative differentiation of avian embryonic hematopoietic progenitors. PMID- 7479488 TI - Eimeria acervulina infection: evidence for the involvement of CD8+ T lymphocytes in sporozoite transport and host protection. AB - Host-parasite interactions during coccidial infection are complex, and host responses leading to parasite elimination are not well understood. Interactions between host cells and parasites were investigated using two-color immunofluorescent staining of duodenal tissue infected with Eimeria acervulina. Tissues were removed from TK strain chickens 24, 48, and 72 h after primary infection with 2.5 x 10(7) normal or irradiated oocysts and also following secondary infection with 2.5 x 10(7) normal oocysts. The tissues were embedded in freezing compound, frozen, sectioned, and fixed in cold acetone. Sections were stained with monoclonal antibodies to identify CD4+, CD8+, Ig+, and Bu1a+ lymphocytes, as well as macrophages and parasites. At 24 h after all infections, sporozoites were seen primarily in CD8+ lymphocytes and macrophages. Many CD8+ lymphocytes were seen at all times in infected tissues and frequently appeared to be in contact with infected epithelial cells. Sporozoites were occasionally seen in CD4+ lymphocytes but none were noted in Ig+ or Bu1a+ lymphocytes. Meront development was extensive after primary infection with normal oocysts, sparse after primary infection with irradiated oocysts, and moderate after secondary infection. PMID- 7479489 TI - Tolerance of turkeys to diets high in trypsin inhibitor activity from undertoasted soybean meals. AB - The objectives were to determine the responses of turkeys to soybean meals (SBM) differing in urease and trypsin inhibitor activity, to estimate the AME of diets containing these SBM, and to determine the responses to supplemental L-Met and L Lys. Four experiments were conducted with poults 1 to 3 wk of age and one with turkeys 6 to 8 wk of age. In Experiment 1, the trypsin inhibitor activities (TI) were 1.8, 4.2, 5.4, 7.0, and 8.8 mg trypsin inhibited/g SBM (method of Hamerstrand et al., 1981). The corresponding urease indices were .02, .14, .51, .90, and 1.5 pH units. The SBM were 46% of the diet. Significant pancreatic hypertrophy occurred with dietary concentrations of TI of 3.2 mg/g and above. At 4.0 mg TI/g of diet, the feed:gain ratio was increased, but body weight gain and AME of the diet were reduced. In Experiments 2, 3, and 4, poults responded similarly to Met additions to diets containing 46% SBM with TI of 1.8 or 4 mg/g SBM, or to Met or Met plus Lys additions to diets containing 40.7 or 49.6% SBM with TI of 2 or 11 mg/g SBM. In Experiment 5, the SBM contained TI at 4.3, 6.1, 8.9, or 12.5 mg/g. The corresponding urease indices were .05, .27, 1.43, and 1.72 pH units. The SBM were 49.6% of the diet. Using 6 to 8 wk old turkeys, the AME of the four diets were determined to be 2.76, 2.71, 2.58, and 2.57 Mcal/kg. The AME of diets containing 4.4 and 6.2 mg TI/g of diet were reduced (P < .05). In conclusion, through 3 wk of age, turkeys can tolerate soybean TI concentrations of 2.5 mg TI/g of diet. Turkeys 6 to 8 wk of age can tolerate 3 mg of soybean TI/g of diet. PMID- 7479490 TI - Dates and date pits as ingredients in broiler starting and coturnix quail breeder diets. AB - Two broiler growth and two quail breeder studies were conducted to evaluate the dietary potential of dates, date meat, and date pits. The test ingredients were incorporated in broiler starting diets at levels ranging from 8 to 43% dates, 16 to 43% date meat, and 5 to 27% date pits. The quail breeder diet had ingredient ranges of 10 to 30% dates, 8 to 24% date meat, and 5 to 15% date pits. The date ingredient diets supported broiler weights and feed conversions comparable to or better than the control diet. Quail breeder feed consumption decreased at the 30% date level; however in the second trial 24% dates supported normal intake. Egg production and egg weight on all date and date part diets were equal to those of the control. PMID- 7479491 TI - Replacing protein in corn-soybean turkey diets with methionine and lysine. AB - The extent to which dietary protein can be minimized by using Met and Lys supplements for market turkeys has been investigated in three experiments involving 2,750 birds. Large White (Nicholas) male turkey poults were fed corn soybean meal diets of varying protein level to supply Lys from 80 to 120 of NRC (1984) from day-old to 18 wk of age. Performance with diets of 90% NRC Lys was equal to that with diets of higher Lys, provided that SAA were at 100% of NRC. Up to .2% Lys.HCl could be substituted into the 100 or 90% of NRC diets with no depression in performance, suggesting that the other essential amino acids were present in adequate amounts. When turkeys were fed diets of 85% of NRC (1984) Lys, maximum growth and breast meat yield were obtained with supplemental SAA at 100% during 0 to 18 wk of age and Lys at 100% during 12 to 18 wk of age. The requirements for other amino acids were supplied by diets formulated to provide 85% of the Lys requirement from protein. These values were for turkeys subjected to 18 C during final growout; they were not adequate in warmer temperatures. PMID- 7479492 TI - Comparison of dietary 25-hydroxycholecalciferol and cholecalciferol in broiler chickens. AB - We conducted a series of 10 feeding trials involving over 36,000 broilers. The effects of various dietary levels of cholecalciferol (vitamin D3) or 25 hydroxycholecalciferol (25-OH-D3) were compared using a "basal" dosage level of 69 micrograms/kg feed, as well as levels ranging from .5 to 1.5 times the basal level. For all 10 studies, average body weight increased by an average of .042 +/ .03 kg (P < .001) and adjusted feed efficiency decreased (improved) by an average of .026 +/- .0046 kg/kg (P < .001) in birds fed 25-OH-D3 in comparison to those fed vitamin D3 at the basal level. Changes in mortality were not detected. Evaluation of different dietary levels of 25-OH-D3 revealed a significant dose response relationship, with maximal effects on weight gain, feed efficiency, and breast meat yield being observed in the range of 50 to 70 micrograms/kg feed. Preliminary studies with different levels of vitamin D3 suggested no additional benefits on weight gain or feed efficiency with higher dietary levels of vitamin D3. Serum 25-OH-D3 concentrations increased more rapidly in birds fed 25-OH-D3 than in birds fed vitamin D3. There were significant correlations with body weight, feed conversion, and serum 25-OH-D3 concentrations, with no correlations observed between serum 1,25-(OH)2D3 concentrations and these variables. PMID- 7479493 TI - Dietary formulation with cottonseed meal on a total amino acid versus a digestible amino acid basis. AB - The results of previous experiments indicated that performance of chicks fed corn cottonseed meal (CSM) diets was inferior to performance of chicks fed a corn soybean meal (SBM) diet and that true digestibilities of amino acids (AA) in CSM were lower than those in SBM. Therefore, the current study evaluated the formulation of diets containing expander-solvent CSM on an equivalent total AA basis vs an equivalent digestible AA basis compared to a corn-SBM diet. True digestibility of AA in the CSM, corn, and SBM were determined using the precision feeding assay, with cecectomized roosters and TMEn was determined using conventional roosters. One-week-old male chicks were then fed diets containing corn and SBM only or diets containing increasing levels of CSM from 5 to 40% that were formulated to be equal in total or digestible AA to the corn-SBM diet. All diets contained 20.7% CP and 3,200 kcal TMEn/kg. Weight gain and feed efficiency of chicks fed 20% or more CSM on a total AA basis were less (P < .05) than those of chicks fed the corn-SBM diet. Growth performance of chicks fed up to 20% CSM on a digestible basis was equivalent to that of chicks fed the corn-SBM diet, but dietary levels of 30 or 40% CSM depressed performance even on a digestible basis. Increasing most essential AA by 15% in the 40% CSM (digestible basis) diet had no effect on performance. True digestible AA and ME values determined in chicks were similar to those determined in roosters. These results indicate that formulation of diets containing CSM on a digestible AA basis is superior to formulation on a total AA basis and that at least 20% CSM could be fed to chicks if the diet contains adequate levels of digestible AA and Fe to bind gossypol. The results also indicated that the depression in performance from feeding 30 or 40% CSM diets was not associated with dietary AA levels. PMID- 7479494 TI - The effect of enzyme predigestion on the quality of poultry by-product meal from whole turkey mortality. AB - Enzyme predigested poultry by-product meal made from onfarm, whole turkey mortality (E-TBM) was compared with soybean meal (SBM) and commercial poultry by product meal (CPBM) in each of three experiments. In Experiment 1, young poults were fed diets containing 16, 20, or 24% CP with either SBM or E-TBM as protein sources. A separate group of poults was fed a nitrogen-free diet. The E-TBM was higher than SBM in protein, fat, lysine, methionine, and total sulfur amino acids. Diets containing E-TBM resulted in increased BW, feed consumption, and a higher protein efficiency ratio (PER) and net protein ratio (NPR) compared with diets containing SBM. In Experiment 2, feeding nonenzyme-treated turkey by product (TBM) or E-TBM resulted in significantly improved BW and feed efficiency compared with CPBM but this may have been due to decreased protein in diets containing CPBM. Protein source did not affect feed intake, PER, NPR, or net protein utilization (NPU). In Experiment 3, the TMEn content of CPBM was lower than both TBM and E-TBM but lysine and methionine digestibilities were greater. PMID- 7479496 TI - Ontogeny of pituitary and serum growth hormone in growing turkeys as measured by radioimmunoassay and radioreceptor assay. AB - Some mammalian studies have revealed a wide discrepancy in pituitary and circulating growth hormone (GH) measurements determined by immunological and biological assay methods. Recent studies demonstrating that avian GH exists in numerous isoforms raise concerns that immunological measurement of GH may not accurately reflect the amount of biologically active hormone present. We sampled eight different male turkeys of a commercial strain weekly until 6 wk of age, and then biweekly until 20 wk. Total pituitary GH content and serum GH concentration were measured by avian GH RIA and radioreceptor assay (RRA). The highest mean serum GH concentration occurred at 3 wk, and the ontogeny of serum GH content from 1 to 8 wk was similar whether measured by RIA or RRA. Pituitary GH content was highest at 6 wk, but RIA and RRA estimates differed markedly throughout the study. Pituitary content of biologically active GH, as estimated by RRA, exceeded that of immunologically active GH from 2 to 10 wk, whereas the reverse was true at 14, 18, and 20 wk. We conclude that this avian GH RIA accurately measures bioactive circulating turkey GH, but that the pituitary of the young turkey may contain bioactive GH isoforms that have poor immunological activity in our RIA. PMID- 7479495 TI - Viability assessment of turkey sperm using fluorescent staining and flow cytometry. AB - Turkey sperm viability was evaluated using several fluorescent stains both singularly and in combination. Dilution curves and several extenders were used to determine optimal stain concentrations. Semen was collected from eight toms, pooled, diluted, stained, and evaluated microscopically within 2 h of collection. Replicates were assessed for both viable and nonviable sperm (green and red fluorescence, respectively) using flow cytometry. SYBR-14, which likely requires membrane potential for optimal fluorescence, or Calcein AM (CAL), which assesses the membrane integrity of cells (both green fluorescence), in combination with propidium iodide (PI) to stain the dead or degenerating cells (red fluorescence) provided optimal results. Sperm were killed by unprotected freeze-thawing to provide mixed aliquots containing known amounts of fresh:killed sperm. The percentage of viable sperm, as determined by SYBR-14 with PI or CAL with PI staining, were 76.6, 58.8, 39.3, 20.1, 8, and 73.5, 55.8, 36.0, 17.1, .4, respectively, for ratios of 100:0, 75: 25, 50:50, 25:75, and 0:100 of fresh:killed mixtures. Semen from 30 individual toms was collected on 2 d and examined in replicate using these staining combinations. The proportion of viable sperm, as indicated by uptake of SYBR-14 or CAL stain, ranged from 55.8 to 86.7 and 38.0 to 86.1, respectively. Staining combinations were effective in estimating the viability of turkey sperm and could be useful for monitoring sperm viability before and after storage. PMID- 7479497 TI - Evidence of a ventilation-perfusion mismatch during acute unilateral pulmonary artery occlusion in broilers. AB - A ventilation-perfusion (V/Q) mismatch can be diagnosed when a moderate increase in cardiac output (CO) causes systemic hypoxemia, thereby exposing a pulmonary incapacity to fully oxygenate the additional blood flowing through the pulmonary vasculature. The susceptibility of broiler chickens to hypoxemia was evaluated in lightly anesthetized, clinically healthy, 40- to 49-d-old males. A snare placed around one pulmonary artery permitted acute, reversible shunting of the entire CO through the unobstructed lung. Blood samples were withdrawn from arterial and venous cannulas for blood gas analysis, and a pulse oximeter was used for noninvasive measurements of the percentage saturation of hemoglobin with oxygen in arterialized capillary beds. The partial pressure of oxygen in arterial blood averaged 103 mm Hg during control periods, 79 mm Hg (hypoxemia) when the pulmonary artery snare was tightened for 12 min, and 101 mm Hg within 5 min after releasing the snare. The percentage saturation of hemoglobin with oxygen before, during, and after tightening the snare averaged, respectively, 96, 91, and 96% for arterial blood, 81, 55, and 78% for venous blood, and 87, 67, and 88% for arterialized capillary beds. Tightening the snare increased the partial pressure of carbon dioxide and the hydrogen ion concentration above control levels in both arterial and venous blood, and these variables returned to control levels upon release of the snare. The combined data constitute direct evidence that clinically healthy broiler chickens are susceptible to hypoxemia during an acute moderate (approximately twofold) increase in pulmonary blood flow. PMID- 7479498 TI - A comparison of argon, carbon dioxide, and nitrogen in a broiler killing system. AB - Argon, CO2, and N2 gasses were each evaluated in a broiler chicken gas killing system. Birds were killed by individual exposure to one of the three gasses for 2 min in a flow-through system. The gasses were evaluated by determining the time, in seconds, for the following responses: first reaction to the gas, loss of posture, eye closure, initiation of death struggle, and cessation of respiration. Percentage blood loss over a 3-min bleed time was determined by weight loss. Breast muscle pH values were determined at 15 min and 24 h post-mortem on the Pectoralis major muscle. Carbon dioxide resulted in the earliest first reaction, loss of posture, eye closure, and initiation of struggle. Argon and N2 exhibited a delayed first reaction, a less severe early reaction, but a more severe unconscious death struggle. All birds died in approximately 75 s. Results indicate that the flow-through gas system takes longer to kill broilers than the immersion systems previously reported. Gas killing resulted in lower (P < .05) blood loss. Initial breast muscle pH values were significantly highest for the birds killed with CO2, followed by the control treatment, which was significantly higher than the values for broilers killed with either Ar or N2. After 24 h of chilling, there were no differences in broiler breast muscle pH among the four treatments. These results indicated that a flow-through gassing chamber may be a feasible, although slower, method of performing gas killing as compared to an immersion system. PMID- 7479499 TI - Detection of Salmonella enteritidis in eggs and chicken with enzyme-linked immunosorbent assay. AB - An ELISA previously developed for the rapid detection of Salmonella enteritidis (SE) in environmental samples was modified and applied to food samples. A sandwich ELISA was designed that employs affinity-purified rabbit polyclonal antibodies for the capture stage and highly specific monoclonal antibodies for the detection stage. Thirty-nine species of bacteria other than SE, including 32 Salmonella species, were included in cross-reactivity testing with ELISA. Results showed no reactivity with any species tested besides SE. Salmonella enteritidis was added to homogenized food samples (chicken skin, meat, and eggs) to test ELISA sensitivity. The lower limit for ELISA detection of SE was 10(4) cells/mL for pure cultures and in 10% meat (wt/vol), 10(5) cells/mL in 10% skin (wt/vol), and 10(7) cells/mL in 10% eggs (wt/vol). Salmonella enteritidis detection with ELISA was confirmed with the Food and Drug Administration (FDA) Bacteriological Analytical Manual (BAM) method. Results were obtained within 24 h for ELISA method compared to 96 h for the BAM procedure. Results show that sensitivity of ELISA can vary with the type of food tested for detection of SE. PMID- 7479500 TI - Heritability of ascites and the ratio of right to total ventricle weight in broiler breeder male lines. AB - The objectives of this study were to induce ascites in three pure male line populations by subjecting them to cold stress, and to calculate heritability estimates for ascites (ASCITES) and the ratio of right to total ventricle weight (RATIO) in these populations under cold stress. Data were collected during the winter of 1993 to 1994 from 3,436 cockerels representing three commercial broiler breeder male lines. The male lines used in this study were characterized by: rapid growth rate and good feed efficiency (RG), moderate growth, good conformation, and excellent livability (MG), and maximum white meat yield and rapid growth (YD). Birds were cold stressed and euthanatized at 8 wk of age. Ratios of right to total ventricle weight and ascites (birds with fluid in the abdominal cavity) were recorded on every bird. Line means for the incidence of ascites were 17.5, 18.7, and 33.5% for RG, MG, and YD, respectively. Lines RG and MG did not differ from each other for ascites, but both were less than YD (P < .05). Means and standard deviations for the ratio of right to total ventricle weight (RATIO) were .299 +/- 0.9, .297 +/- .08, and .294 +/- .09 for RG, MG, and YD, respectively. Line means for RATIO did not differ. Heritability estimates of RATIO were .21 +/- .09, .21 +/- .09, and .27 +/- .08 for RG, MG, and YD, respectively; and heritabilities of ascites were .36 +/- .10, .11 +/- .08, and .44 +/- .09, respectively. Phenotypic correlations of ASCITES with RATIO were .54, .43, and .50 in RG, MG, and YD and genetic correlations of ASCITES with RATIO were .69 +/- .13, .46 +/- .33, and .78 +/- .10 in RG, MG, and YD, respectively. PMID- 7479501 TI - Restricted maximum likelihood estimates of genetic parameters of adult male and female Rhode Island red chickens divergently selected for residual feed consumption. AB - In adult chickens, feed intake can be predicted by multiple linear regression from body weight, change in body weight during the recording period (for males and females), and egg mass (for females). Residual feed consumption (RFC) is estimated by the deviation of observed from predicted values for feed intake. A divergent selection experiment has been conducted since 1975 in a Rhode Island Red population. Each sex was selected on the basis of its own RFC, Line R+ for high values of RFC and Line R- for low values. In addition to the traits measured to obtain RFC, egg production traits and body measurements (BM) have been recorded, including wattle length, shank length, and rectal temperature. After pooling data from the two lines and the base population, the data set included 1,064 males, each with 7 variables, and 3,780 females, each with 11 variables. Genetic parameters were estimated by a multivariate derivative-free-restricted maximum likelihood procedure, which yields estimates free of bias due to selection and inbreeding. The RFC appeared to be moderately heritable in males (h2 = .33) and in females (h2 = .27) and poorly correlated between sexes, with a genetic correlation of .19 between RFC in males (RFCm) and in females (RFCf). The RFCm and RFCf were weakly correlated with egg production traits. Significant, positive correlations were found between RFCm and the BM traits that were related to heat dissipation. Correlations between feed intake, body weight, and BM traits recorded in males or in females were always < 1, except for shank length.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7479502 TI - Random amplified polymorphic DNA comparisons among broiler lines selected for incidence of tibial dyschondroplasia. AB - Lines selected for high (H) and low (L) incidence of tibial dyschondroplasia (TD) for eight generations and a randombred control (C) line of broiler chickens were fingerprinted by random amplification of genomic DNA mixed from 20 individuals of each line with 20 oligonucleotide primers. Among these 20 primers, 15 could distinguish the H from the L line, 14 the H from the C line, and 13 the L from the C line. Band sharing (BS), on the average over 20 primers, was .7 for the H vs L comparison and .8 for both H vs C and L vs C comparisons. The levels of BS calculated from individuals was .6 between the H and L line, .7 between the H and C line, and .7 between the L and C line. The ranking of BS values obtained from individual DNA samples was consistent with that obtained from the mixed DNA samples. Genomic distance between divergently selected lines (H vs L) was larger than that between the divergently selected lines and randombred line (H vs C and L vs C). Individual variation within lines was detected in spite of eight generations of selection. Results showed that eight generations of divergent selection for TD incidence in broiler chickens had resulted in genetic variation among lines. The procedure of random amplified polymorphic DNA assay using mixed DNA samples could be used to evaluate genetic distance among lines of chickens. PMID- 7479503 TI - Correlated responses to long-term divergent selection for eight-week body weight in chickens: growth, sexual maturity, and egg production. AB - Thirty-six generations of divergent selection for BW at 8 wk of age (BW8) resulted in approximately an eightfold difference between the high (HWS) and low (LWS) lines for this trait. Correlated traits included BW at 4, 24, and 38 wk of age (BW4, BW24, BW38, respectively), age at first egg (AFE), and percentage hen day egg production (HDP). Responses of BW4 followed the same pattern as that for the selected trait, with the response about five times greater during the first 18 than the last 18 generations of selection in Line HWS and less than two times greater in Line LWS. For BW24 and BW38, correlated responses were greater for LWS than for HWS females without feed restriction, suggesting changes in growth curves after selection age. Although AFE was delayed in both lines, the delay was greater in Line LWS (some individuals of which were anorexic) than in Line HWS and greater in the second half than the first half of the experiment. For pullets that commenced lay, HDP declined slightly in both lines. Correlations between BW at 4, 8, 24, and 38 wk of age were moderate to high and positive in both lines. When feed intake was restricted in Line HWS, however, there were no correlations of BW4 or BW8 with BW24 or BW38. Correlations between AFE and BW at all ages were negative in Line LWS. In Line HWS there were negative correlations of AFE with BW24 and with BW38. Relaxed lines, established periodically during the experiment, were satisfactory monitors of environmental influences for primary and correlated traits. PMID- 7479504 TI - Turkey breeder hen performance by strain during consecutive lay periods. AB - The results of this study provide information about reproductive performance among commercially available strains of Large White Turkey breeder hens, Nicholas (N), British United (B), and Hybrid (H). The hens were managed identically through two consecutive lay periods, summer-fall (1st-yr) followed by winter spring (recycled). Effects of the intensity (351 vs 24 lx) of supplemental light were evaluated during the 1st-yr lay period. Data were collected for onset of oviposition, egg production, BW, feed consumption, livability, egg weight, and egg components. Light intensity had similar effects on all variables measured. There were neither strain differences nor genetic interactions with light intensity treatment. There was no difference in livability among strains (P = .20) or between year of lay (P = .08), and there was no strain by year interaction. Body weight differed by strains and year and there was a significant strain by year interaction. In general, N were heavier than H or B in the 1st-yr and early recycle lay period, whereas H and B were similar in BW in both lay periods. Recycled hens of all strains were heavier than 1st-yr hens. The onset of lay was earlier in H than B or N in both lay periods and all three strains had a delay in onset of lay after recycling. Through 23 wk photostimulation the B hens produced more eggs per hen than N or H hens (B > N > H) and recycled hens produced more eggs per hen than 1st-yr hens. Strain, year, and strain by year interaction effects were significant for egg weight. The first eggs laid, as well as subsequent eggs in both lay periods, were heaviest in N hens. In all strains, percentage yolk increased and albumen decreased with time in lay. However, N hens had a smaller percentage yolk and greater albumen than H or B hens. The mean percentage shell was greatest in H hens. PMID- 7479505 TI - The effect of early feed restriction on broiler performance. AB - In an attempt to determine the effect of early feed restriction on broiler performance, nine trials were conducted. Feed restriction was based on previous 24-h feed consumption values of the full-fed control groups. Feed restriction was from either 7 to 14 d of age (Trials 1 to 5) or 8 to 16 d of age (Trials 6 to 9). The treatments for Trials 1 to 5 were 90, 75, and 60% of previous 24-h feed consumption of full-fed controls. The treatments for Trials 6 to 9 were 80 and 60% of previous 24-h feed consumption of full-fed controls. Early feed restriction did not affect abdominal fat. Early feed restriction did improve feed conversion (grams of feed:BW). Males and females equated at 41 d of age for an 8% reduction (90% feed restriction) at 14 d of age when compared with full-fed control broilers. Neither females or males could overcome a 17 or 18% weight reduction (75% feed restriction) at 14 d of age by 41 d of age. At 48 or 49 d of age, both males and females could equate in weight with full-fed control broilers for a weight reduction of 27 and 31% (60% feed restriction) at 16 d of age. Mortality or deformed legs did not differ between any of the comparisons. PMID- 7479506 TI - Consumer acceptance of irradiated poultry. AB - A simulated supermarket setting (SSS) test was conducted to determine whether consumers (n = 126) would purchase irradiated poultry products, and the effects of marketing strategies on consumer purchase of irradiated poultry products. Consumer preference for irradiated poultry was likewise determined using a home use test. A slide program was the most effective educational strategy in changing consumers' purchase behavior. The number of participants who purchased irradiated boneless, skinless breasts and irradiated thighs after the educational program increased significantly from 59.5 and 61.9% to 83.3 and 85.7% for the breasts and thighs, respectively. Using a label or poster did not increase the number of participants who bought irradiated poultry products. About 84% of the participants consider it either "somewhat necessary" or "very necessary" to irradiate raw chicken and would like all chicken that was served in restaurants or fast food places to be irradiated. Fifty-eight percent of the participants would always buy irradiated chicken if available, and an additional 27% would buy it sometimes. About 44% of the participants were willing to pay the same price for irradiated chicken as for nonirradiated. About 42% of participants were willing to pay 5% or more than what they were currently paying for nonirradiated chicken. Seventy-three percent or more of consumers who participated in the home use test (n = 74) gave the color, appearance, and aroma of the raw poultry products a minimum rating of 7 (= like moderately). After consumers participated in a home-use test, 84 and 88% selected irradiated thighs and breasts, respectively, over nonirradiated in a second SSS test. PMID- 7479507 TI - Effects of feeding fumonisin B1 present in Fusarium moniliforme culture material and aflatoxin singly and in combination to turkey poults. AB - Diets containing 200 mg fumonisin B1/kg of feed and .75 mg aflatoxin/kg of feed singly or in combination were fed to female turkey poults (Nicholas Large White) from day of hatch to 21 d of age. When compared with controls, 21-d body weight gains were reduced 10% by fumonisin B1, 39% by aflatoxins, and 47% by the combination. Relative weights (grams/100 g body weight) of the kidney and pancreas increased in poults fed the diet containing aflatoxins alone, whereas the relative weight of the liver decreased. Relative weights of the liver and pancreas increased in the poults fed the fumonisin diet. Relative weights of the kidney, pancreas, and gizzard increased in the poults fed the combination diet, whereas the relative weight of the liver decreased. Most serum constituents, hematology values, and activities of enzymes measured were altered in poults receiving the diets containing aflatoxins with or without fumonisin B1. No major histological lesions were observed in tissues from control poults or poults fed the diet containing fumonisin alone. Lesions associated with aflatoxins were only observed in the liver and occasionally in the kidney of poults fed the diets containing aflatoxins with or without fumonisin B1. The primary hepatic change was bile duct hyperplasia with some hepatocellular degeneration and necrosis and megalocytosis. Occasional necrotic and degenerating tubular epithelial cells were observed in the kidneys. The increased toxicity in poults fed the combination diet for most variables can best be described as additive, although some variables showed less than additive toxicity.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7479508 TI - Chicken tumor necrosis-like factor. I. In vitro production by macrophages stimulated with Eimeria tenella or bacterial lipopolysaccharide. AB - HD11, a transformed avian macrophage cell line, and chicken peripheral blood leukocyte-derived macrophages (PBL-M phi) were stimulated with bacterial endotoxin lipopolysaccharide (LPS) or Eimeria tenella sporozoites and merozoites. The specific cytotoxicities of the culture supernatants against different target cell lines were measured, and the kinetics of tumor necrosis-like factor (TNF) production by HD11 and PBL-M phi were also measured. The results showed that HD11 and PBL-M phi secreted a TNF-like factor when stimulated with Eimeria parasites or LPS. A time- and dose-dependent TNF-like factors production by PBL-M phi was observed poststimulation with Eimeria parasites. Chicken TNF-like factor preferentially kills CHCC OU-2 cells, a fibroblast cell line of chicken origin, when compared to LM cells, a murine cell line used for mammalian TNF. This study indicates that chicken M phi produce a significant level of TNF-like factor following coccidial infection. PMID- 7479509 TI - Sodium requirement for broiler breeder hens. AB - Two experiments were conducted with Arbor Acres hens to determine their Na requirement. Sodium chloride was added to a corn-soybean meal diet to furnish daily intakes of 35, 65, 95, 125, and 150 mg per hen in Experiment 1. The 150 mg level was replaced with 180 mg in Experiment 2. The hens were maintained in floor pens with wood shavings used as litter. Experiment 1 was conducted for 9 wk and Experiment 2 for 12 wk. Egg production was significantly reduced during Week 4, when the hens had an intake of 35 mg Na/d. Egg production was reduced by Week 7 when the diet furnished 65 mg/d. Plasma Na was reduced and plasma K increased when the intake was 35 or 65 mg/d. Performance data from 6 to 9, and 6 to 12 wk were used in Experiments 1 and 2, respectively, to determine the daily Na requirement. The daily requirement for maximum production was 113.8 and 96 mg and the requirement for egg mass was 105 and 100 mg in Experiments 1 and 2, respectively. PMID- 7479510 TI - Endogenous amylase levels and response to supplemental feed enzymes in male turkeys from hatch to eight weeks of age. AB - Amylase and xylanase enzyme concentrations in the pancreas, small intestine, and crop were measured in Nicholas male poults fed diets with and without supplemental amylase and xylanase from 0 to 8 wk of age. Eight birds from each of three diets (control, amylase-supplemented, xylanase-supplemented) were killed every 3 d to determine the amylase and xylanase activity within the pancreas, small intestine, and crop. Pancreatic organ weight was not affected by diet, indicating an absence of dietary amylase effect upon pancreatic tissue growth. Pancreatic amylase activity was not consistently affected by diet. Amylase activity within the intestinal chyme increased sporadically with dietary amylase supplementation over the control and xylanase-supplemented diets. Increasing supplemental amylase activity levels may provide more conclusive evidence of an additive effect of dietary amylase and endogenous amylase activity. Xylanase supplementation within the feed did not affect endogenous amylase activity. PMID- 7479511 TI - Effects of protein level and enzyme supplementation upon growth and rate of digesta passage of male turkeys. AB - An experiment was conducted to determine the effect of feeding enzyme supplements (Avizyme and protease) at two levels of dietary crude protein (24 and 28%) to male turkeys from 0 to 5 wk of age. The Avizyme-protease enzyme mixture was fed at five concentrations within each protein level in an effort to determine an optimal level of supplementation. Enzymes were added to the 24% protein diet to determine whether the supplements would enhance the growth performance of birds fed the lower protein diet to the level of those fed the 28% protein diet. Rate of passage was evaluated to determine whether an ANF effect corresponded to enzyme concentration or protein level. Dietary protein level significantly influenced growth. Birds fed the 28% protein diet had improved body weight gain, feed consumption, and feed efficiency of 11.5, 6.5, and 4.4%, respectively, when compared with birds fed the 24% protein diet. Enzyme addition to the 24% protein corn-soybean meal diet produced a graded response but did not improve growth over the control. Although enzyme supplementation improved poult utilization of the 24% protein diet, growth and feed utilization were not equal to the performance of the poults fed the 28% protein diet. When the enzyme mixture was added to the 28% diet, performance was not consistently altered. Rate of digesta passage was not different between the levels of dietary protein or among levels of enzyme supplementation, indicating that the applied protein levels did not deleteriously affect nutrient utilization. PMID- 7479512 TI - Growth and intestinal morphology of male turkeys as influenced by dietary supplementation of amylase and xylanase. AB - Three diets, a control and two diets supplemented with an enzyme cocktail premix containing either amylase or xylanase, were each fed to 100 male poults (10 replicates of 10 poults per pen) from 0 to 5 wk of age to observe the effects upon body weight gain, feed intake, and feed efficiency. The amylase-supplemented diet significantly increased feed efficiency through the first 2 wk and significantly increased body weight gain and feed intake through the first 3 wk. Xylanase supplementation did not improve growth or feed efficiency over the control. Mean villus length within the jejunum and ileum was significantly increased at 2 and 3 wk of age by dietary supplementation of amylase when compared with the control and xylanase diets. These findings suggest that the increased growth associated with the amylase diet during 0 to 3 wk can, in part, be explained by the increase in absorptive surface area, allowing for increased digestion of available nutrients coupled with increased enzyme activity for carbohydrate degradation from the supplemental enzymes. PMID- 7479513 TI - The nutritive value of dietary fats in relation to their chemical composition. Apparent fat availability and metabolizable energy in two-week-old chicks. AB - Broiler chicks at 2 wk of age were fed the following fats and oils at 4% in a corn-wheat-soybean basal diet: palm oil (PO); tallow (T1 and T2); tallow + soybean oil (50:50, TSO); tallow+acidulated soybean oil soapstock (50:50, TASO); soybean oil (SO); and linseed oil (LO). All of these, with the exception of T1, contained 5% soybean lecithin. The apparent fat availability (AFA) and AME of added fats ranged from 78.8 to 96.0% and from 7.32 to 8.95 kcal/g, respectively. Three fat sources (TSO, SO, and LO) resulted in the highest values for these two measurements, whereas PO, T1, T2, and TASO showed the lowest values. There was no difference for AME of diets among fat-supplemented treatments, regardless of the difference in the nutritive values of the fats and oils tested. The apparent availability of each fatty acid (AAFA) for C16:0, C18:0, C18:1, C18:2, and C18:3 was calculated from the analysis of the diets and excreta collected over a 4-d period. The results of the present study indicated that 2-wk-old chicks utilize TSO, SO, and LO more efficiently than PO, T1, T2, and TASO. PMID- 7479514 TI - Metabolizable energy and amino acid availability of full-fat seeds, meals, and oils of flax and canola. AB - The TME, true amino acid availability (TAAA), AME, and apparent amino acid availability (AAAA) were determined in full-fat seeds, meals, oils, and meal-oil mixtures of flax and canola using Single Comb White Leghorn roosters by the force feeding and total excreta collection assay. The metabolizable energy values were corrected to zero nitrogen balance to obtain AMEn and TMEn. The AMEn and TMEn contents of canola seed (4.46 and 4.56 kcal/g) were respectively higher (P < .05) than those of flax seed (3.75 and 3.75 kcal/g). Flax meal contains contents of AMEn (2.07 kcal/g) and TMEn (2.07 kcal/g) similar to those (1.98 and 2.09 kcal/g) of canola meal. There were no differences in AMEn and TMEn values between flax oil (8.10 and 8.28 kcal/g) and canola oil (8.25 and 8.46 kcal/g). No differences were observed among AME, AMEn, TME, and TMEn values within each test ingredient except canola meal: its TME value was higher (P < .05) than its AMEn value. The TAAA values were higher for full-fat seeds than for the meals and also higher for canola seed and meal than for flax seed and meal. The AAAA values, although lower than TAAA values, followed similar patterns as TAAA values. Both AAAA and TAAA values were higher when the ingredients were fed with a basal diet than when fed alone. PMID- 7479515 TI - Re-evaluation of the methionine and protein requirements of the broiler breeder hen. AB - Three experiments were conducted to study the Met and protein requirements of Arbor Acres broiler breeder hens. Broken line regression analysis in Experiments 1 and 3 indicated the Met requirement was 335 and 323 mg/d, respectively, with a production of a daily egg content (EC) of 42.3 or 43.0 g. A broken line could not be fitted to the data in Experiment 2. Therefore, this procedure could not be used to calculate a requirement. An average of 7.83 mg Met was required to produce 1 g of EC. Using this value, the requirement for the three experiments would be 344 mg at the level of EC produced in these experiments. The requirement would be higher with either a higher rate of lay or with larger eggs. PMID- 7479516 TI - Effect of alpha-tocopherol on antioxidants, lipid peroxidation, and the incidence of pulmonary hypertension syndrome (ascites) in broilers. AB - Research has demonstrated a compromised antioxidant capacity in broilers with pulmonary hypertension syndrome (PHS). Thus, the objective of the present study was to assess the effects of vitamin E on PHS-induced mortality, tissue antioxidants, and plasma lipid peroxides in male broilers. Control broilers were provided normal ventilation but others, maintained under low ventilation conditions to induce PHS, were randomly assigned to nonimplanted (NI), placebo (PL), or vitamin E (VE) implanted groups. The VE implant released a total of 15 mg of alpha-tocopherol from 0 to 3 wk of age. Tissues and blood samples were obtained at 3 and 5 wk of age from birds with (PHS+) and without (PHS-) PHS. Five week PHS cumulative mortality was lowered by alpha-tocopherol with mortality rates of 3.6, 4.2, 11.9, and 11.8%, for Controls, VE, NI, and PL groups, respectively. The PHS+ birds exhibited lower body weights, higher hematocrit, right ventricular hypertrophy, lower alpha-tocopherol and glutathione (GSH) concentrations in liver and lung, as well as indicators of oxidative stress, including elevated plasma lipid peroxides and lower oxidized GSH in liver and erythrocytes, at 5 wk of age. All birds exhibited lower erythrocyte catalase activity at 5 than at 3 wk of age. An improved antioxidant capacity was observed in VE birds, including higher liver and lung alpha-tocopherol at 3 and 5 wk, higher liver GSH at 3 wk, and lower plasma lipid peroxide values at 5 wk of age. Direct correlations observed between body weight and plasma lipid peroxides at 3 wk (r = .45) and between right ventricular hypertrophy and plasma lipid peroxides at 5 wk (r = .48), suggests that lipid peroxidation plays a role in the etiology of PHS. The results indicate that the VE implant was effective in lowering PHS induced mortality in broilers apparently by attenuating processes leading to lipid peroxidation. PMID- 7479517 TI - Dietary cholesterol and fat saturation effects on plasma esterified and unesterified cholesterol in selected lines of Japanese quail females. AB - Three lines of Japanese quail females, randombred controls (CL), high response (HL), and low response (LL) lines, selected for plasma total cholesterol for 18 generations, were fed all-plant-source, nonatherogenic diets to which 0 or .5% cholesterol were added from 6 to 18 wk of age. In all three lines, plasma cholesterol increased when cholesterol was fed; however, responses were greater in the HL than in the LL line, with CL intermediate. In a second experiment, females of the three lines were fed, from 6 to 14 wk of age, four isocaloric, isonitrogenous plant-source diets to which were added: 1) 10% glucose monohydrate (cerelose); 2) 10% cerelose + .1% cholesterol; 3) 4% corn oil; or 4) 4% coconut oil. Baseline data obtained before feeding experimental diets indicated that the HL had significantly higher plasma total, esterified (EC) and unesterified (UEC) cholesterol than LL and that nonovulating females had higher concentrations of esterified cholesterol than ovulating females. Diets used did not affect cholesterol fractions in the ovulating females, although there were significant differences among lines. Dietary cholesterol significantly increased the ratio of EC to UEC. Sclerotic lesion scores were higher in the HL than the LL birds and in birds fed the coconut oil diet. PMID- 7479518 TI - Film oxygen transmission rate effects on ground chicken meat quality. AB - Effects of the packaging film oxygen transmission rate (OTR) on the odor, color, aerobic plate count, and cooked volatile compounds in ground chicken leg meat were evaluated over a 14-d refrigerated storage (4 C) period. Freshly desinewed and ground chicken leg meat was packaged under minimal vacuum in five films of different OTR. Films ranged from 30 to 12,000 mL oxygen/m2 per 24 h. Odor scores of meat in packages opened after 10 d were lower for the lowest OTR film than films with higher OTR. Aerobic plate counts increased at a faster rate in the higher OTR films, and cooked meat volatile profiles showed little variation due to OTR film type. Specific products will require different packaging to optimize shelf-life quality. For ground chicken leg meat, an intermediate OTR film is best to maintain a majority of the quality attributes during refrigerated storage. PMID- 7479520 TI - The effect of chopped computer and bond paper mixed with wood shavings as a litter material on broiler performance. AB - The impact of utilizing chopped computer and bond paper as a litter material on growth performance of broilers reared to market age was evaluated. In the first experiment, broilers were placed in pens at 10 birds per square meter, and four different litter treatments were used; 1) 100% chopped computer and bond paper (CCP); 2) a mix of 75% CCP and 25% wood shavings (WS); 3) 50% CCP and 50% WS; 4) 100% WS. Experiment 2 evaluated the same treatments, but with densities of 14 birds per square meter. Litter treatments had no effect on body weight gain, feed consumption, conversion, or mortality for either of the two experiments. No differences were found for percentage litter moisture among treatments. Chopped computer and bond paper alone or mixed in different percentages with WS can be used as an alternate litter material for rearing broilers as long as there is good waterer management, especially with open watering systems. These systems may have the tendency to increase the incidence of litter caking. PMID- 7479519 TI - Yolk thiobarbituric acid reactive substances and n-3 fatty acids in response to whole and ground flaxseed. AB - n-3 fatty acid (FA)-enriched shell eggs have been proposed as an economical source for increasing consumption of n-3 FA. As dietary n-3 FA sources vary with respect to fatty acid profile and lipid stability, the comparison of resulting egg products is warranted. This study was designed to determine yolk thiobarbituric acid reactive substances (TBARS) and n-3 FA when hens were fed whole or ground flaxseed. Gold flaxseed (whole or ground) fed at levels of 5 or 15% were compared to a 1.5% menhaden oil or a typical control layer ration. A 5 wk feeding trial was used with 21 hens per treatment. Eggs were collected and FA composition determined during Weeks 4 and 5 of the feeding trial. The TBARS content of yolk was also determined. All flaxseed treatments increased total n-3 FA (C18:3 + C20:5 + C22:6) marine n-3 FA (C20:5 + C22:6) did not increase proportionately with increasing dietary flaxseed. Seed form did not influence n-3 FA deposition at the 5% level of flaxseed; however, ground 15% flaxseed resulted in greater total n-3 FA deposition than whole 15% flaxseed. Yolk TBARS were not different due to diet, indicating that seed form did not influence the oxidative quality of these n-3 FA-rich lipids. PMID- 7479521 TI - Therapeutic potential of drugs with mixed 5-HT4 agonist/5-HT3 antagonist action in the control of emesis. AB - Drugs interacting with serotonin (5-hydroxytryptamine, 5-HT) receptors are of value in the treatment of several gastrointestinal disturbances. Selective 5-HT3 receptor antagonists (ondansetron, granisetron, tropisetron) are widely utilized to control emesis induced by chemotherapy and radiation, while agonists at 5-HT4 receptors (cisapride, renzapride, BIMU compounds) are endowed with gastrointestinal prokinetic action. Here we overview the therapeutic potential of drugs with potent mixed 5-HT4 agonist/5-HT3 antagonist properties (i.e. BIMU 1) in the management of anticancer therapy-induced emesis and of intestinal adynamic post-operative conditions associated with vomiting. In the former situation, the agonism at 5-HT4 receptors is expected to be of benefit via two possible mechanism: (i) inhibition of 5-HT release from enterochromaffin cells; (ii) restoration of anally driven peristaltic waves in the upper gastrointestinal tract. Moreover, 5-HT4 receptor-induced prokinetic activity may counteract colonic constipation, an unwanted effect which occurs in a number of patients treated with pure 5-HT3 receptor antagonists. Additionally, the above mentioned drugs might be of value in post-operative conditions associated with intestinal adynamia and emesis sensitive to 5-HT3 receptor blockade. PMID- 7479522 TI - Behavioural assessment in rats of the antipsychotic potential of the potent dopamine D2 receptor antagonist, (-)eticlopride. AB - The effects of the selective D2 DA receptor antagonist, (-)eticlopride, a drug belonging to the benzamide class, were investigated on the D2 DA agonist SND 919- and CQP 201-403-induced stereotyped behaviour and on CQP 201-403-induced shaking, in rats, and on isolation-induced aggression, in mice. (-)Eticlopride was also tested over a wide dose range (5-1200 micrograms kg-1, s.c.) for sedative and cataleptic activity, in rats. For comparison, some experiments were performed with (-)sulpiride (10 and 40 mg kg-1, s.c.) The data obtained show that ( )eticlopride differs from (-)sulpiride and potentially modifies animal behaviour, whether spontaneous or induced; moreover, they suggest a potential clinical use for this neuroleptic in the management of psychotic states. PMID- 7479524 TI - Formation of F2-isoprostanes in oxidized low density lipoprotein: inhibitory effect of hydroxytyrosol. AB - Oxidatively-modified low-density lipoproteins (LDL) contribute to the onset of the atherosclerotic disease. A recently discovered marker of lipid peroxidation in a series of prostaglandin F2-like compounds, the prostaglandin isomers isoprostanes, that are generated from arachidonic acid through cyclooxygenase independent pathways following free radical injury and are endowed with potent biological activities. The incidence of cardiovascular disease in the Mediterranean area is low, possibly because of the type of fat (mainly olive oil) and other components (e.g. fruits and vegetables) of the diet. Natural antioxidants abound in this kind of diet and may also contribute to the observed protection from coronary heart disease (CHD) by retarding the formation of the atherosclerotic plaque. Olive oil, the major dietary fat in the Mediterranean countries, is rich in phenols with antioxidant properties. We thus investigated the formation of isoprostanes during in vitro LDL oxidation and tested the effect of an olive-oil-extracted phenol (i.e. hydroxytyrosol). Our data show that production of isoprostanes and other markers of lipid peroxidation occurs during LDL oxidation and is inhibited by hydroxytyrosol. PMID- 7479523 TI - Novel met-enkephalin analogue: a potent systemic mu agonist antinociceptive agent. AB - A new met-enkephalin analogue (compound 82/205) was evaluated for its opioidergic activity in mice. The compound showed antinociception (warm water tail-flick test), tolerance, cross tolerance to morphine and physical dependence. The time course of antinociceptive effect of the compound was comparable to morphine. The antinociceptive ED50 (mumol kg-1, i.p.) values for the compound and morphine base were 5.31 and 7.59, respectively. Its antinociceptive effect was blocked by naloxone, beta-FNA (mu antagonist) and naloxonazine (mu1 antagonist) but not by ICI 174,864 (delta antagonist). Naloxone precipitated withdrawal jumpings were 2.6 times less in compound 82/205 treated mice than the morphine treated group. The new analogue compound 82/205 is a potent mu agonist antinociceptive with a possible weak dependence liability. PMID- 7479525 TI - Tumour necrosis factor mediates E-selectin production and leukocyte accumulation in myocardial ischaemia-reperfusion injury. AB - The aim of our study was to examine the mechanism of E-selectin production and leukocyte accumulation in myocardial ischaemia-reperfusion injury. Myocardial injury was induced in anaesthetized rats by the clamping of the left main coronary artery followed by reperfusion. After thoracotomy a silk suture was placed under the left coronary artery. The ligature was tied for a period of 1 h and after this period it was untied and the ischaemic myocardium was reperfused for 1 h (MI/R rats) or removed (SHAM MI/R rats). Myocardial ischaemia plus reperfusion in untreated rats decreased survival rate, produced a marked myocardial necrosis, enhanced cardiac myeloperoxidase activity (a marker enzyme commonly used to assess polymorphonuclear leukocyte infiltration) and increased serum creatinephosphokinase (CPK) activity, serum levels of tumour necrosis factor-alpha (TNF-alpha) and serum levels of soluble E-selectin (sE-selectin). Furthermore, MI/R rats had an increased pressure rate index studied as a quantitative means for assessing myocardial oxygen demand. Administration of cloricromene, an inhibitor of TNF-alpha, reduced TNF-alpha production, significantly lowered serum sE-selectin levels, blunted leukocyte accumulation in the ischaemic myocardium and protected the myocardium from injury due to ischaemia and reperfusion. The results of the present study show an involvement of E-selectin in vivo in the pathogenesis of myocardial ischaemia and reperfusion and suggest that TNF-alpha may induce in vivo the production of a specific adhesion mechanism which sustains leukocyte infiltration. PMID- 7479526 TI - Atypical antagonism of D1-receptor-mediated vasodilator response in the perfused kidney by SCH23390. AB - Vasodilator responses to dopexamine, fenoldopam and dopamine, which are known to have agonist activity at D1-dopamine receptors, were examined in the rat isolated perfused kidney preparation. Perfusion pressure was raised by perfusing with the thromboxane TxA2 analogue, U46619, and vasodilator responses were observed as dose-related falls in perfusion pressure. Propranolol (10(-6) M) and prazosin (10(-6) M) were present throughout to eliminate beta 2 and alpha 1-adrenoceptor mediated responses, respectively. The vasodilator responses were antagonized by SCH23390 (10(-9) M), indicating that they were mediated via D1-receptors. The displacements of the dose-response curves for fenoldopam, dopexamine and dopamine were, however, non-parallel with significant depression of the maxima to 30.2, 37.9 and 34.3%, respectively. In the presence of SCH23390 (10(-8) M) and prazosin (10(-6) M), dopexamine, isoprenaline and noradrenaline produced dose-related renal vasodilation. This was antagonized by propranolol indicating a role for beta-adrenoceptors. In the case of dopexamine, the maximum response was depressed in the presence of propranolol. The reason for the atypical blockade of vasodilator responses by SCH23390 was investigated. One possibility was the appearance of transient vasoconstrictor responses at higher doses of fenoldopam, dopexamine and dopamine, usually preceding the vasodilatation. The possibility was therefore considered that the vasoconstriction may have opposed the usual vasodilation at high doses and thus limited the size of the maximum vasodilation in the presence of SCH23390. The vasoconstriction by fenoldopam was not antagonized by S-sulpiride, the D2-receptor antagonist but was blocked by mianserin and therefore attributed to 5-HT2 receptor stimulation.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7479527 TI - Effect of (-)-cathinone, a psychoactive alkaloid from khat (Catha edulis Forsk.) and caffeine on sexual behaviour in rats. AB - The effect of (-)-cathinone, caffeine and their combinations was studied on the sexual behaviour of male rats. Male sexual activities were assessed by recording the erectile responses (grooming of genitalis, yawning/stretching and homosexual mounting), in the absence of females. The copulatory behaviour was observed by caging males with receptive females brought into oestrus with s.c. injection of oestradiol benzoate and progesterone. The copulatory pattern of male rats (mounting, intromissions, ejaculations and refractory period) was recorded. The oral treatment of cathinone (5 mg kg-1 day-1), caffeine (50 mg kg-1 day-1) and their combinations for 15 days increased arousal (motivation) in male rats as evidenced by increased mounting performance and anogenital investigatory behaviour. However, erectile and ejaculatory responses, measured in the present study, showed no stimulant effect. It is conceivable from the present results that cathinone, the psychostimulant constituent of khat modified masculine pattern behaviour and caffeine also changed the effect of cathinone when administered concomitantly. However, our data provide no evidence that cathinone could be considered as an aphrodisiac. PMID- 7479528 TI - Hepatic glutathione after ethanol administration in rat: effects of cimetidine and omeprazole. AB - As a fraction of ingested ethanol (EtOH) is metabolized by gastric mucosa, different amounts of alcohol reach the liver, when the same dose is administered by oral or intravenous route. In previous experiments, we demonstrated that the decrease of hepatic reduced glutathione (GSH) is less pronounced and is followed by a quicker recovery after oral than after intraperitoneal administration of the same amount of EtOH. Therefore, the time-course of hepatic GSH concentration seems to be an indirect assay of EtOH metabolism by the liver. On the basis of these findings, any condition causing a reduced function of gastric alcohol dehydrogenase (ADH) should show up as a more severe depletion of hepatic GSH. In the same rat experimental model we determined the effects of cimetidine and omeprazole administration on gastric ADH activity and on the time-course of hepatic GSH after EtOH load. Cimetidine was shown to inhibit gastric ADH with a Ki of 0.167 +/- 0.009 mmol l-1; accordingly, the pretreatment with this drug (20 mg kg-1 b.w. per day for 1 week) determined, after oral EtOH load, a marked reduction of hepatic GSH, likewise after intraperitoneal administration. Omeprazole exerted only a marginal inhibition on gastric ADH and this drug (0.3 mg kg-1 b.w. per day for 1 week) did not modify the time-course of hepatic GSH concentrations after EtOH load. This study indicates that the inhibition of gastric ADH, when associated with EtOH intake, induces depletion of the hepatic GSH concentration and, therefore, possible liver damage. PMID- 7479529 TI - Effects of prostaglandin E1, and papaverine on non-neurogenic and neurogenic contraction of the isolated rabbit erectile tissue. AB - Experiments were performed to get further insights into the erectogenic mechanism of prostaglandin E1 (PGE1), which was compared to that of papaverine (PAP). PGE1 and PAP were effective in abolishing the contraction induced by N-ethylmaleimide (NEM), an adenylate cyclase blocker. However, preincubation with PGE1 but not with PAP markedly attenuated the amplitude of adrenergic nerve mediated contraction following prolonged electrical field stimulation. Preincubation with PGE1 was ineffective in counteracting the increase in tension due to exogenous norepinephrine. These data together with previous studies corroborate the hypothesis that in the presence of PGE1 a dual erectogenic mechanism takes place in modulating the cyclic-adenosine-monophosphate metabolism of the cavernous smooth muscle cell as well as the release of norepinephrine from the sympathetic terminal. PMID- 7479530 TI - Enhancement effect of carteolol on the clonidine-induced vasodilation of rat mesenteric arteries. AB - It has demonstrated that carteolol can increase the endothelium-dependent vasodilation induced by alpha-2 adrenergic agonist. In order to evaluate the effect of carteolol, and to clarify the mechanism, we examined the effects of 10 microM carteolol on the vasodilation induced by increasing doses (10(-7)-10(-4) M) of clonidine in perfused rat mesenteric arteries preconstricted with 100 microM phenylephrine. Clonidine elicited a dose-dependent vasodilation of the mesenteric arteries preconstricted with phenylephrine. Carteolol enhanced the vasodilation induced by higher doses (10(-5) and 10(-4) M) of clonidine, although carteolol itself exerted no direct vasodilating effect. On the other hand, 10 microM propranolol or 10 microM metoprolol did not augment the clonidine-induced vasodilation. In the presence of 100 microM NG-monomethyl L-arginine (LNMMA), an analogue of L-arginine, the enhancement of the clonidine-induced vasodilation by carteolol was abolished. This inhibition by LNMMA was restored with 300 microM L arginine, but not with 300 microM D-arginine. These results suggest that carteolol enhances the clonidine-induced vasodilation by an endothelial-related mechanism mediated by the release of endothelium-derived nitric oxide in resistance vessels. PMID- 7479531 TI - Biochemical changes induced by pyrphenoxone in the lens of rabbits and rats. AB - The xanthomatine analogue, pyrphenoxone, which is known to diminish the incidence of cataract in animals and in man, was applied in two different in vivo models of cataract induced in rabbits by tryptophan-free dietary regimen and in rats by hypergalactosemic diet. The drug was also applied at different concentrations in an in vitro model of cataract. It was found that soluble proteins and sulphurated amino acids of the lens in all in vivo and in vitro models of cataract were higher after pyrphenoxone was applied. Furthermore, the drug treatment was followed by a dose-dependent increase in reduced glutathione content in the lens of rabbits and rats. The same was found in the in vitro model of cataract. These results suggest that pyrphenoxone may act by inducing various biochemical changes that lead to a protection of lens against oxidative processes. PMID- 7479532 TI - 1st European Congress of Pharmacology. Milan, Italy, June 16-19, 1995. Abstracts. PMID- 7479533 TI - Been there, done that, what's next? PMID- 7479534 TI - Application as scholarship: a community client experience. AB - A community health nursing experience with a complex aggregate client was developed for B.S.N. completion students at a private university. Working in collaboration with the neighborhood group, a survey based on Spradley's notion of healthy neighborhoods and Cottrell's idea of community competence was developed. Community health nursing students drew a representative sample of neighborhood dwellers and interviewed them to assess their concerns. Data were summarized, inferences drawn, and problems amenable to nursing intervention identified and prioritized. Students presented their findings to the neighborhood group, city service providers, and various volunteer agencies. This experience provided service to the community client, public service for faculty, and an exceptional learning opportunity for students. PMID- 7479535 TI - Abuse during pregnancy: frequency, severity, perpetrator, and risk factors of homicide. AB - To determine the frequency, severity, and perpetrator of abuse during pregnancy as well as the occurrence of risk factors of homicide, a stratified, prospective cohort analysis was completed on 1,203 African-American, Hispanic, and Anglo women in urban public health prenatal clinics. All women were assessed for abuse at the first prenatal visit and twice again during the pregnancy. All women were administered the Conflicts Tactics Scale, the Index of Spouse Abuse and the Danger Assessment Scale. Prevalence of physical abuse during pregnancy was 16%, one in every six women. Women abused during pregnancy had significantly higher scores on all instruments and more risk factors of homicide when compared with women abused prior to but not during pregnancy. To protect women's safety, a protocol of assessment and intervention for abuse must be standard care for all pregnant women. PMID- 7479536 TI - Women with diabetes during pregnancy: sociodemographics, outcomes, and costs of care. AB - This study provides sociodemographic, outcome, and cost data on a population (N = 55) of predominately low-income, diabetic women who were hospitalized during pregnancy. Study findings indicated that 43 percent received no prenatal care in the first trimester, 20 percent delivered a low-birthweight infant, 47 percent had a cesarean delivery, and 63 percent reported an annual income under $12,500. Following the women's initial admission for glucose control, 19 acute care visits and 32 rehospitalizations were recorded for them. The mean hospital charges for antepartum initial hospitalization for glucose control were $4,665 (4.3 days). The mean charges for postpartum hospitalization were $7,793 (4.3 days). The mean hospital charges per infant were $12,991. Given the data presented in this study, it is imperative that monies be targeted to provide a broad spectrum of health care services that will meet the unique needs of this population. These services should address not only the needs related to superimposed disease state but also identify mechanisms to assist women to receive care prior to conception, or at the very least to begin prenatal care in the first trimester of pregnancy. PMID- 7479537 TI - Evaluation of public health nursing home visits to pregnant women on WIC. AB - Selected pregnant women participating in the Special Supplemental Food Program for Women, Infants and Children (WIC) were receiving public health nursing (PHN) home visiting services. The purposes of this study were to describe PHN services provided to pregnant women on WIC; and to compare birth outcomes and prenatal care (PNC) utilization between two groups, women with WIC services only and women with WIC plus PHN services. In this retrospective cross-sectional study, the sample consisted of all women on WIC (N = 398) at the Madison Department of Health who delivered singleton births during 1991, divided into the WIC-only group (n = 301) and the WIC + PHN group (n = 97). Three instruments were used for data collection: birth certificates, the WIC record, and the antepartal nursing record. Data were described; differences between groups were tested. Primary PHN activities included: assessment of physical and behavioral indicators, parenting skills, and support systems; teaching about nutrition, use of drugs, danger signs, labor signs, and stress management; and referrals to a variety of community resources. No difference in adequacy of PNC or in birth outcomes was found between the two groups. Findings have been used to plan the agency's future PNC coordination services. PMID- 7479538 TI - Client characteristics associated with not keeping appointments for public health nursing home visits. AB - Home visits have been identified as an intervention strategy for high-risk pregnant women. A necessary component of this intervention strategy is for the women to be home for the visit with the nurse. The purpose of this study was to identify factors associated with women not keeping their home visit appointments with a public health nurse. Subjects were 232 low-income high-risk pregnant women who received at least one home visit by a public health nurse. Younger age, more stressful life events, receipt of medical assistance, inadequacy of prenatal care, noncompliance with health recommendations, and longer gestational time were associated with not keeping public health nursing appointments. While this study provides some insight into the factors associated with women not keeping their appointments with the public health nurse, further investigation is warranted to identify strategies to reduce the incidence of not-kept appointments. PMID- 7479539 TI - Building community capacity for health promotion: a challenge for public health nurses. AB - The centennial celebration of public health nursing is a strong reminder of the tradition and practice of public health nursing's commitment to communities. Partnerships with communities give public health nurses fiduciary responsibility to be actively involved in public health reform to advance health promotion and health protection. Public health nurses must rise to the challenge to build community capacity through facilitating community participation, enhancing community health services, and coordinating public policy to achieve core public health responsibilities of assessment, policy, and assurance. This paper explicates strategies for building community capacity for health promotion. PMID- 7479540 TI - Health promotion and the "ideology of choice". AB - Assessment and intervention in public health nursing are intricately linked to health promotion. Health promotional strategies in turn focus attention on life style modifications. A broad base of research warns of the dangers of the "ideology of choice" that currently accompany such lifestyle-change attempts. The "ideology of choice" is inherent in pervasive views that individuals are "responsible for" and "choose" their disease. The rhetoric of individual responsibility pervades the discourse around both health promotion and health care reform. Without continual awareness of both the complex and multifaceted nature of such life-style "choices," and the balancing of a compassionate stance toward human frailty, health promotion attempts easily degenerate into a victim blaming stance. Public health nurses need to be particularly aware of these dangers during this period when cost constraints are generating more pervasive "ideology of choice" rhetoric, targeting those with the scarcest resources and the most profound needs. PMID- 7479541 TI - HIV-related concerns and educational needs of public health nurses in a rural state. AB - An anonymous questionnaire was completed by 369 nurses in public health departments in a rural Southeastern state to examine the relationship between nurses' prior HIV training and their HIV-related knowledge, attitudes, concerns, and perceived training needs. The survey was conducted in three predominantly urban counties with the highest number of AIDS cases and in 38 rural counties with two or fewer reported AIDS cases. Knowledge answers were generally 70%-90% correct and attitudes more favorable than unfavorable. Attitude was more frequently associated with HIV training level than was knowledge. Concerns about working with persons with high-risk behaviors were expressed by more than half the nurses and were more prevalent in rural areas. Nurses with more training had more concerns about client care and fewer fears about HIV work. Almost all (85%) were concerned about lack of community resources. Most nurses wanted more training of the client-sensitive type provided by the state. With the increasing incidence of HIV/AIDS in rural areas, planning continuing education for staff not only on new developments and current therapies (desired by 98%) but on managing feelings about clients with high-risk behaviors seems especially important not only for the staff, but for their significant others and communities. PMID- 7479542 TI - Chattanooga Creek: case study of the public health nursing role in environmental health. AB - Public health nurses have two primary roles in protecting their communities from hazardous substances: community assessment and health education. Developing assessment skills in environmental health enables public health nurses to collaborate with other federal, state, and county agencies in identifying public health hazards and making health-based recommendations at hazardous waste sites needing remedial or removal interventions. Community health education empowers communities to minimize their exposure to hazardous wastes in their environment. Methods for community environmental health assessment and interventions are demonstrated in this article by activities conducted at the Chattanooga Creek site in Chattanooga, Tennessee. A thorough assessment and collaborative approach between government agencies, local health professionals, and community members resulted in a successful community health education program and this site's placement on the National Priorities List. PMID- 7479543 TI - Women's beliefs concerning condom acquisition and use. AB - Condoms are a time-honored and reliable method of protection against pregnancy and sexually transmitted infections. However, their use, and thus their effectiveness, is determined by individual behavior. The purpose of this paper is to report attitudes and salient beliefs related to condom use in a sample of adult women. The study used Ajzen and Fishbein's Theory of Planned Behavior to identify modal, salient beliefs regarding condom acquisition and use as intentional behaviors. The study sample consisted of 58 community women who reported using condoms for contraceptive purposes within the last five years. In face-to-face, audiotaped interviews, open-ended questions were used to solicit beliefs regarding condom acquisition and use. All subject narratives were content analyzed for recurrent themes. Women cited accessibility and effectiveness in preventing pregnancy and sexually transmitted diseases as both advantages and as factors contributing to the ease of acquisition and use. Disadvantages and factors that might deter condom acquisition and use included embarrassment, objections by male partner, and effect on spontaneity. Overall, subjects exhibited accurate knowledge regarding the benefits of condom acquisition and use. However, it is possible that expressed negative beliefs could take precedence in decision-making and reduce the probability of consistent condom use. PMID- 7479544 TI - Developing a collaborative community partnership program in medical asepsis with tattoo studios. AB - The possibility of transmission of infectious agents during tattooing has become a legitimate issue of concern for health care providers. A collaborative educational program was developed by a county health department, College of Nursing, and tattoo artists to address issues of medical asepsis with the goal of producing a mechanism for certification of tattoo studios. The group's effort was enhanced by recognizing each other's value systems and by the mutual need for a successful program. A framework for developing, implementing, and evaluating community partnerships was addressed. This program demonstrated that community health nurses can play an instrumental role in collaborating with both health care providers and personal-service workers to minimize transmission of infectious agents during cosmetic procedures. PMID- 7479545 TI - The effect of pH and temperature on the self-association of recombinant human interleukin-2 as studied by equilibrium sedimentation. AB - The self-association of recombinant human interleukin-2 (rhIL-2) in solution was investigated as a function of pH and temperature using equilibrium sedimentation. Studies were performed at pH 3.6, 6.5 and 8.2, at 1 degree C and 20 degrees C. A model assuming an ideal single molecular species describes the data observed at pH 6.5 at both temperatures. At pH 8.2, the data from both temperatures can be better described by a weak monomer-dimer association equilibrium. The values of the association constants obtained indicate the presence of less than 10% dimer at a concentration of 1 mg/ml at both temperatures. At pH 3.6, aggregates with a Z average molecular weight of over 35 times that of monomeric rhIL-2 were formed. The smallest associating species present under these conditions corresponds to the monomer, which produces aggregates with a wide range of molecular weights. The monomer appears to be in equilibrium with the smallest aggregates, in that a model describing an indefinite association fits the data obtained at the highest centrifugal speed. No model was found to successfully describe the association of the monomer into the much larger aggregates observed at lower speeds. This may be the result of the lack of rapid thermodynamic reversibility of the larger aggregates. Temperature was found to have no significant effect on the largest aggregates that were formed at pH 3.6. PMID- 7479546 TI - Interdomain interactions in the chimeric protein toxin sCD4(178)-PE40: a differential scanning calorimetry (DSC) study. AB - The thermal denaturation of the chimeric protein toxin known as sCD4(178)-PE40 (sCD4-PE40) was studied using differential scanning calorimetry (DSC). sCD4-PE40 consists of HIV-binding domains of the T-cell membrane protein known as CD4 and the cytotoxic domains of Pseudomonas exotoxin A (PE40). sCD4-PE40 undergoes two DSC transitions. An endothermic transition associated with unfolding of the CD4 and PE40 components occurs at approximately 46 degrees C in buffered saline at pH 6.5. An exothermic transition associated with precipitation of unfolded protein occurs at higher temperatures. Both transitions are irreversible. DSC studies of solutions of pH 5.0 to 9.5 indicate that sCD4-PE40 shows maximal thermal stability at around pH 6.5. Variable pH experiments are also presented on solutions of sCD4(183) and PE40 revealing how these components denature as independent structural entities. sCD4(183) denaturation occurs at significantly higher temperatures than does the CD4 component of sCD4-PE40. PE40 denaturation occurs at the same temperatures as sCD4-PE40. These results suggest that the native CD4 and PE40 components are independent and non-interacting entities in the chimeric sCD4-PE40 molecule and that unfolding of the less-stable PE40 component induces unfolding of the CD4 component. These destabilizing interdomain interactions of sCD4-PE40 are in contrast to the stabilizing interactions which apparently exist in wild-type Pseudomonas exotoxin A between its PE40 domains and the cell binding domain of the native toxin (analogous to the CD4 component in sCD4-PE40). Reasons are discussed why the type of interdomain interactions observed for sCD4-PE40 might be the norm for chimeric proteins. PMID- 7479547 TI - Primary interactions of three quaternary ammonium compounds with blastospores of Candida albicans (MEN strain). AB - The absorption of three quaternary ammonium compounds (QAC), cetylpyridinium chloride, cetrimide and benzalkonium chloride, onto the surface of blastospores of Candida albicans (MEN strain) was examined at room temperature. Equilibrium uptake occurred in less than 30 seconds for cetylpyridinium chloride and cetrimide whereas 5 min contact time was required for benzalkonium chloride. The adsorption of all three agents may be mathematically described as Langmuirian and hence a concentration-dependent formation of drug-monolayer on the surface of the blastospore occurred. From this the number of molecules adsorbed onto the surface of a single blastospore was calculated to be 1.33 x 10(12), 3.17 x 10(12) and 2.32 x 10(12) for cetylpyridinium chloride, cetrimide and benzalkonium chloride, respectively. These dissimilarities are most likely due to differences in the orientations of both the cationic nitrogen atom and the accompanying lipophilic portions of each QAC at the blastospore surface. Relating these observations to the known antiadherence effects of cetylpyridinium chloride and cetrimide, it can be concluded that monolayer coverage of the blastospore surface with QAC does not account for the observed reduced adherence. This suggests that the anti-adherence effects are due to either direct interaction with, or steric blockade of, adhesions on the blastospore surface. PMID- 7479548 TI - Antitumor effect of arterial administration of a medium-chain triglyceride solution of an angiogenesis inhibitor, TNP-470, in rabbits bearing VX-2 carcinoma. AB - Using rabbits bearing VX-2 carcinoma on the inner side of the leg, we examined the antitumor activity of a medium-chain triglyceride (MCT) solution of an angiogenesis inhibitor, TNP-470 (AGM-1470, 6-O-(N-chloroacetylcarbamoyl) fumagillol), following administration into the femoral artery feeding the tumor. The MCT solution of TNP-470 (1 and 5 mg) strongly suppressed tumor growth following a single intra-arterial (i.a.) injection 2 or 3 weeks after tumor inoculation. Moreover, remarkable regression of well-developed tumors, those 4 weeks after inoculation, was obtained by i.a. injection of the MCT solution containing 20 mg of TNP-470 without any influence on body weight. The antitumor effects were potentiated by coadministration of doxorubicin or mitomycin C (MMC) in the solution or microspheres containing MMC. In a shell-less chorioallantoic membrane (CAM) assay, angiogenesis was inhibited when a droplet of the MCT solution containing 25 micrograms of TNP-470 was placed on the CAM for 2 days, suggesting that the prolonged antitumor effect resulted from the inhibition of tumor neovascularization by sustained drug release from the preparation. These results indicate that i.a. injection of the MCT solution of TNP-470 is promising for treating well-developed tumors. PMID- 7479549 TI - Selective inhibition of bitter taste of various drugs by lipoprotein. AB - Previously, we demonstrated that lipoprotein composed of phosphatidic acid (PA) and beta-lactoglobulin (LG) selectively and reversibly suppress the frog taste nerve response to bitter substances. In the present study, we examined the effects of various lipoproteins on the taste sensation to various stimuli in humans by a psychophysical method. Among various lipoproteins composed of different of lipids and proteins, the lipoproteins composed of PA and proteins were most effective in suppressing bitter taste. The lipoproteins composed of PA and LG, bovine serum albumin, ovalbumin, alpha-lactoalbumin or casein similarly suppressed effects on sensation of bitter taste. Using PA-LG, the effects on taste sensation to various stimuli were examined. The bitter taste of all twelve substances examined was inhibited, while saltiness of NaCl and sweetness of sucrose were not inhibited. The inhibition of bitter taste was completely reversible. Masking of the target sites for bitter substances on the taste receptor membranes with PA-LG seems to contribute to the inhibition of bitter taste. Direct binding of the bitter substances to PA-LG in the medium also contributes to the inhibition of bitter taste of certain substances. Among various drugs, basic and hydrophobic substances such as quinine, denatortium and propranolol have low taste thresholds and are said to be the most bitter. PA-LG most effectively suppressed the bitter taste of such substances. PA originates from soybeans and the proteins used except for bovine serum albumin originate from milk or eggs, and hence the lipoproteins can be safely used to mask the bitter taste of drugs. PMID- 7479550 TI - Prolonged blood circulation in rats of nanospheres surface-modified with semitelechelic poly[N-(2-hydroxypropyl)methacrylamide]. AB - Semitelechelic poly[N-(2-hydroxypropyl)methacrylamide]s (ST-PHPMA) containing one amino end-group and differing in molecular weight were synthesized by radical polymerization in the presence of 2-aminoethanethiol (AET) as chain transfer agent. These polymers were covalently attached via amide bonds to the surface of nanospheres based on a copolymer of methyl methacrylate, maleic anhydride, and methacrylic acid. When compared to unmodified nanospheres, those with the surface modified with ST-PHPMA possessed a decreased protein (albumin, IgG, fibrinogen) adsorption in vitro, an increased intravascular half-life as well as a decreased accumulation in the liver after intravenous administration into rats. The higher the molecular weight of the ST-PHPMA, the more pronounced the changes in these properties. The results obtained have clearly demonstrated that covalently attached ST-PHPMA chains are efficient in decreasing the biorecognition of negatively charged (hydrophilic) polymer surfaces. PMID- 7479551 TI - Modelling mucoadhesion by use of surface energy terms obtained from the Lewis acid-Lewis base approach. II. Studies on anionic, cationic, and unionisable polymers. AB - Surface energies of carbopol, chitosan, hydroxypropyl cellulose (HPC) and poly(HEMA) were assessed from contact angle and surface tension experiments. The surface energy was considered in terms of an apolar Lifshitz-van der Waals term and a polar acid-base term, which in turn is divided into electron donor and electron receptor (Lewis acid-Lewis base) contributions. Using these surface energy terms the interaction of dry and hydrated polymer with mucin in the presence of either artificial gastric or intestinal fluid, or saline was predicted. The predictions were related to measured forces of detachment. There was a significant difference between the surface energy on dry and hydrated HPC and also for carbopol; for the other polymers either the surface energy of the hydrated material was not detectable, or the effect of hydration was minimal. There were good correlations between mucoadhesive strength and the calculated free energies of interaction between mucin and polymer in the presence of each of the fluids, for each individual polymer. Thus, two trends were observed, one for unionisable and the other for ionisable polymers. It is argued that the increased mucoadhesion seen with ionisable polymers (compared with the predicted value based on results of unionisable polymers) is a direct result of the ionic interaction. No attempt has been made to correct for the ionisation effect, but the surface energy predictions provide insight into the mechanism of the mucoadhesion process. This approach is useful for understanding and predicting interactions between different materials and biological components. PMID- 7479552 TI - Autoradiographic imaging of the distribution of 5-fluorouracil through cervical tissue following in vitro surface application of a bioadhesive cervical patch. AB - The distribution of 5-fluorouracil through cervical tissue has been assessed following the in vitro application of a bioadhesive patch to excised human cervix. The bioadhesive matrix contained a total of 20 mg of 5-fluorouracil spiked with 5-fluorouracil-6-3H and was applied for fixed periods of either 4 or 24 hours. Tissue slices were sectioned perpendicular to the plane of the applied patch and the autoradiographic image developed by placing a frozen tissue slice on Hyperfilm with subsequent instant thawing and refreezing, the resulting bilayer being maintained at -18 degrees C for 24 hours. The developed image was analysed by scanning densitometry and raster scans were visualised with three dimensional contouring software. The autoradiograms showed darker areas surrounding tissue ducts, suggesting that 5-FU was spilling from the lumen into the surrounding stroma. Transport of 5FU via aqueous channels may thus make an important contribution to the rapid penetration of the drug through the cervical stroma. Three-dimensional autoradiographic images showed that, for a 4-hour patch application, there were areas of relatively low drug concentration within the upper 5 mm of tissue, where CIN lesions can exist in the glandular tissue or cervical crypts. However, extending the application time to 24 hours produced areas of high drug concentration extending throughout this region. PMID- 7479553 TI - Interaction between bile salts and beta-adrenoceptor antagonists. AB - The interactions between beta-adrenoceptor antagonists and bile salts were investigated by microcalorimetry. Nadolol and oxprenolol interactions with dihydroxy salts could be described by a 1:1 interaction model with the thermodynamic parameters indicating that the drugs were incorporated within the bile salt aggregates. This weak interaction was primarily hydrophobic although electrostatic attraction also played a role. Atenolol and metoprolol did not interact with the dihydroxy salts. None of the compounds interacted with trihydroxy bile salts or with salts below their aggregation concentration. Phase separation resulted when propranolol and alprenolol were present in dihydroxy salt solutions above a certain concentration with the interaction being of a hydrophobic and electrostatic nature. The implications of these results on in vivo drug absorption are discussed. PMID- 7479554 TI - Enhanced transdermal delivery of diazepam by submicron emulsion (SME) creams. AB - Diazepam, a lipophilic drug with CNS activity, serves here as a model to investigate the efficacy of SubMicron Emulsion (SME) as a novel transdermal vehicle. Diazepam was formulated in various topical regular creams and SubMicron Emulsion creams of different compositions. The different formulations were applied topically and protection against Pentamethylenetetrazole induced convulsive effects in mice was monitored. The efficacy of Diazepam applied topically in emulsion creams strongly depends on the oil droplet size and to a lesser degree--on the formulation composition and the oil type. Processing medium chain-triglyceride (MCT) emulsion with a high-pressure homogenizer causes a drastic reduction in the droplet size, thereby significantly increasing the transdermal activity of Diazepam. In this case both the high-pressure homogenization and the presence of lecithin, an efficient dispersant, contribute to the effective droplet size reduction of below 1 micron, usually between 100 300 nm. The SubMicron Emulsions as vehicles for transdermal delivery of Diazepam generate significant systemic activity of the drug as compared with regular creams or ointments. Transdermal delivery of Diazepam via SME formulations is very effective, and the activity may reach the range of parenteral delivery. A single application of Diazepam in SME cream to mice skin provides pronounced transdermal drug delivery and prolonged protective activity up to 6 hours. PMID- 7479556 TI - The role of metabolites in bioequivalency assessment. II. Drugs with linear pharmacokinetics and first-pass effect. AB - Simulations were conducted to address the question of whether metabolite data are required for bioequivalence evaluation of immediate release formulations with drugs exhibiting linear pharmacokinetics and first-pass effect. Plasma level-time profiles were generated for parent drug and metabolite using relevant rate constants obtained from a bivariate normal distribution and designated random error. Simulation results showed that the need for metabolite data (Cmax) in the assessment of bioequivalence depends on the relative variability between the absorption process of the drug and first-pass route for metabolite(s). The importance of metabolite Cmax data in the evaluation of rate of availability is clearly demonstrated for drugs with a high degree of intra-subject variation in the first-pass metabolism compared to the absorption process of the drug. Under such conditions, a wider confidence interval was found for the metabolite rather than parent drug. Opposite results were obtained when the intra-subject variance was high for drug absorption relative to first-pass effect. Discrepancies were observed for the scenarios in which the elimination pathway of the metabolite is more variable than the absorption process of the drug. The simulation results were in agreement with real bioequivalence data. It is thus recommended that, in the absence of the information on the relative variability of absorption and first-pass process, both parent drug and metabolite data be included for documentation of bioequivalence, should the metabolite(s) play an important role in the determination of efficacy and safety of the drug. PMID- 7479555 TI - Comparison of intestinal permeabilities determined in multiple in vitro and in situ models: relationship to absorption in humans. AB - In vitro and in situ experimental models that are descriptive of drug absorption in vivo are valuable tools in the discovery of new chemical entities that are bioavailable after oral administration. The specific objective of the study was to compare the intestinal permeabilities obtained in the three absorption models for consistency, and to assess the utility of the models in predicting the fraction of dose absorbed in human studies. The intestinal absorption models that were compared are widely used: the rat in situ single-pass intestinal perfusion system, the rat everted intestinal ring method, and monolayers of human colon adenocarcinoma cell line (CACO-2). The models were compared using small molecular reference compounds, as well as a series of peptidomimetic (PM) analogs. Each model had strong potential for estimating the fraction absorbed. For small organic molecules, excellent correlation was observed when permeabilities from CACO-2 cells and perfusions, or everted rings and perfusions, were compared. Weaker correlation was observed between everted rings and CACO-2 cells. Permeabilities for the set of reference compounds and PMs were positively correlated between any two of the three systems. Variance between correlations for reference compounds and PMs are likely due to structural features and physicochemical properties that are unique to the latter class of compounds. The results support caution in extrapolating correlations based on findings with small organic molecules to the behavior of complex peptidomimetics. Corroboration of permeabilities with two methods of determination is a useful cross-validation of experimental systems, as well as producing a reliable permeability assessment.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7479558 TI - The distribution of oleic acid between salbutamol base drug and different propellant blends. AB - The distribution of oleic acid between Salbutamol base drug and the solvent in metered-dose inhalers (MDI's) has been investigated. The equilibrium surfactant concentration in the drug dispersions has been determined using a colorimetric method. The samples examined contained Salbutamol base drug particles and oleic acid dispersed in different propellant blends of freon 11 and 12. The maximum equilibrium concentration observed depended on the propellant blend used. The propellant blend and the distribution of the surfactant affected the dispersion stability. The effect of the surfactant is illustrated by the adsorption isotherm for oleic acid onto the Salbutamol particles. The results are correlated with zeta-potentials and particle size measurements made on similar systems in order to characterize the properties of surfactant stabilized MDI's. PMID- 7479557 TI - Monoclonal digoxin-specific antibodies induce dose- and affinity-dependent plasma digoxin redistribution in rats. AB - The effect of three monoclonal digoxin-specific antibodies on total and free digoxin plasma disposition was studied in rats in order to determine the role of affinity constant (Ka) and dose. Thirty minutes after digoxin infusion, administration of a stoichiometrical dose of the ICIO, 6C9 and 9F5 IgG (Ka = 6 10(9), 3.1 10(8) and 2.5 10(7) M-1, respectively) resulted in a plasma digoxin increase linearly related to Ka. The mean free plasma digoxin was 0.6 +/- 0.4, 7.8 +/- 3.3 and 43 +/- 22% respectively after 1C10, 6C9, and 9F5 IgG infusion in comparison to 70 +/- 9% in the control group. When the IgG:digoxin ratio increased from 1 to 5, plasma digoxin Cmax and AUCT also increased as a function of both affinity (Ka) and dose (N), but not linearly. The product of NKa defined an immunoreactivity factor that was well fitted to the digoxin redistribution parameters (Cmax and AUCT) by a Hill equation. PMID- 7479559 TI - Pharmacokinetics of oral extended-release dosage forms. I. Release kinetics, concentration, and absorbed fraction. AB - In this study, we derive pharmacokinetic models for oral extended-release (OER) drug products with defined in vivo release kinetics (IVRK) and a compartmental system. Fitting the model to clinical data, we were able to examine the correlation between released and absorbed fractions. Furthermore, we found that absorbed fractions of OER products can be expressed by absorption rate and release duration only. The expression is unchanged in different compartmental systems with the same IVRK, implying that the IVRK drives the pharmacokinetic system of an OER product. The apparent absorption rate constant of an OER product can be estimated by solving an implicit equation using observed concentrations. We also propose a new method for calculating absorbed fractions, which is more accurate than Loo-Riegelman method. Ultimately, these methods may permit optimally designed OER products. PMID- 7479560 TI - Experimental design and efficient parameter estimation in preclinical pharmacokinetic studies. AB - Monte Carlo simulation technique used to evaluate the effect of the arrangement of concentrations on the efficiency of estimation of population pharmacokinetic parameters in the preclinical setting is described. Although the simulations were restricted to the one compartment model with intravenous bolus input, they provide the basis of discussing some structural aspects involved in designing a destructive ("quantic") preclinical population pharmacokinetic study with a fixed sample size as is usually the case in such studies. The efficiency of parameter estimation obtained with sampling strategies based on the three and four time point designs were evaluated in terms of the percent prediction error, design number, individual and joint confidence intervals coverage for parameter estimates approaches, and correlation analysis. The data sets contained random terms for both inter- and residual intra-animal variability. The results showed that the typical population parameter estimates for clearance and volume were efficiently (accurately and precisely) estimated for both designs, while interanimal variability (the only random effect parameter that could be estimated) was inefficiently (inaccurately and imprecisely) estimated with most sampling schedules of the two designs. The exact location of the third and fourth time point for the three and four time point designs, respectively, was not critical to the efficiency of overall estimation of all population parameters of the model. However, some individual population pharmacokinetic parameters were sensitive to the location of these times. PMID- 7479561 TI - Gradient high performance liquid chromatographic assay for degradation products of adinazolam mesylate in a sustained release tablet formulation. AB - A gradient high performance liquid chromatographic method was developed to determine degradation products of adinazolam mesylate in a sustained release tablet formulation. Sample preparations were chromatographed on a YMC-Basic column using a formate buffer/acetonitrile gradient with absorbance detection at 254 nm. Adinazolam mesylate was found to degrade at high relative humidity and temperature to form a major product, the 6-aminoquinoline analog, plus numerous other compounds. Five of these compounds were identified and their structures indicate that the solid-state degradation of adinazolam, in the presence of sufficient moisture, involves not only a hydrolytic mechanism, but also an oxidative mechanism. Potential process impurities were resolved from the drug and degradation products. Recovery was near 100% over the 0.5 to 10% range for the major degradate (6-aminoquinoline) and over the 0.5 to 1% range for the other analytes. The method was applied to tablet samples stressed at high relative humidity and temperature. The relative standard deviation of the assay for the 6 aminoquinoline was less than 2% and less than 13% for the minor components. Calculated mass balances (sum of adinazolam plus degradation products in the degraded tablet divided by the same sum in the undegraded tablet) were less than 100% and were dependent on the extent of degradation in the tablet. The average mass balance result obtained for samples that were an average of 9.5% degraded was 95.0 +/- 1.5%. It is possible that the decrease in mass balance with increase in percent degradation may be explained by the formation of many components at trace levels due to degradation by various permutations of hydrolytic and oxidative reaction pathways. PMID- 7479562 TI - Quantitative microdialysis for studying the in vivo L-DOPA kinetics in blood and skeletal muscle of the dog. AB - In this study the microdialysis technique, using alpha-methyldopa as internal standard (IS), is introduced for the in vivo determination of L-DOPA, dopamine (DA), and their metabolites dihydroxyphenylacetic acid (DOPAC) and 3-O-methyldopa (3-OMD) in blood plasma and skeletal muscle extracellular fluid (ECF), in anaesthetised beagle dogs, after i.v. administration of L-DOPA. In a first calibration experiment, the in vivo relative losses (RL) of the compounds and the IS were determined. These were lower in skeletal muscle than in blood plasma. K was defined as the ratio of the RL of the IS to the RL of the compound of interest and was shown to be constant for a certain compound within one tissue. However, except for DA, a significant difference was seen in K values between blood plasma and skeletal muscle. In a second step, the method was validated in blood plasma. The AUC0-->3 values for the non-protein bound L-DOPA did not differ significantly between the dialysis (141.3 +/- 16.0 nmol.h/ml) and traditional whole blood sampling (145.3 +/- 18.7 nmol.h/ml), confirming that microdialysis combined with accurate calibration is a reliable technique for studying the kinetics of drugs in vivo in different tissues. PMID- 7479563 TI - The electrophoretic mobility of tripeptides as a function of pH and ionic strength: comparison with iontophoretic flux data. AB - Capillary electrophoresis (CE) is an extremely efficient separations tool which can also be used to determine fundamental molecular parameters, e.g., the electrophoretic mobility of a molecule. We have studied the changes in the CE estimated electrophoretic mobility of thyrotropin releasing hormone (TRH) as a function of pH and ionic strength. Further, we have used CE to estimate the mobilities of two synthetic analogs of TRH to examine the behavior of positive (basic) and negative (acidic) peptides under the conditions of this work. These data were then compared with literature values of iontophoretic flux of these molecules under similar formulation conditions. Our results suggest that CE could potentially assist formulation optimization for the iontophoretic delivery of peptides. PMID- 7479564 TI - Analysis of diclofenac and four of its metabolites in human urine by HPLC. AB - An HPLC method for the determination of diclofenac (DCF) and four of its metabolites (3'-hydroxydiclofenac, 4'-hydroxydiclofenac, 5-hydroxydiclofenac, and 3'-hydroxy-4'-methoxydiclofenac) in human urine is described. Following base hydrolysis, the samples were neutralized and extracted. Evaporated extracts were reconstituted in mobile phase containing ascorbic acid, and chromatographed, using flow-rate programming, on a reversed-phase column. Absolute recovery (average), was at least 78% for diclofenac and ranged from 75 to 85% for the four metabolites. Standard curves showed linearity over the range of concentrations of 0.2 to 40 ug/mL, using 0.25 mL of urine. Specificity was demonstrated by examining chromatograms of extracts of blank urine from 8 volunteers and 24 study subjects. Good accuracy was observed for all compounds over the concentration range of 0.2 to 40 ug/mL using 0.25 mL of urine. Based on accuracy and precision criteria, the limit of quantitation for all 5 analytes was 0.4 ug/mL, using 0.25 mL of urine. Analysis of urine from subjects with normal and reduced renal function who received diclofenac orally demonstrated that total diclofenac and metabolites excreted in the urine represented approximately 31% and 4% of an oral dose of diclofenac, respectively. PMID- 7479565 TI - Prodrugs to improve the oral bioavailability of a diacidic nonpeptide angiotensin II antagonist. AB - DMP 811 is a diacidic angiotensin II antagonist. It has relatively low oral bioavailability in rats. A prodrug approach to improving oral bioavailability was tested. Five esters were synthesized and their stability in rat plasma in vitro was determined. The hydrolysis rates of these five esters ranged from almost immediate to negligible. A simple n-propyl ester was hydrolyzed very slowly (< 10% in 24 hr) in rat plasma in vitro, and after oral dosing in rats plasma prodrug concentrations were much greater than DMP 811 concentrations. A pivaloyloxymethyl ester (1) was hydrolyzed relatively rapidly in rat plasma in vitro. Prodrug 1 was rapidly hydrolyzed by the intestine in vitro, and the intestinal permeation of DMP 811 was increased. DMP 811 oral bioavailability was 47% in rats dosed with 10 mg/kg 1, compared to 11% for rats dosed with 10 mg/kg DMP 811. However, DMP 811 bioavailability was only 27% after a 2 mg/kg dose of 1. In vitro plasma hydrolysis of 1 was highly species-dependent, with a half-life of 13 hr in human plasma but only 1 min in rat plasma. The prodrug approach has potential for improving the oral bioavailability of DMP 811, but selection of the optimal prodrug must be done in humans or in a species, such as dogs, with hydrolysis characteristics closer to humans. PMID- 7479566 TI - Electrolyte-induced changes in glass transition temperatures of freeze concentrated solutes. AB - Addition of electrolytes to solutions of non-crystallizing solutes can cause a significant decrease in the glass transition temperature (Tg') of the maximally freeze-concentrated solution. For example, addition of 2% sodium chloride to 10% solutions of dextran, PVP, lactose, and sucrose causes a decrease in Tg' of 14 degrees to 18 degrees C. Sodium phosphate has a smaller effect on Tg' and is unusual in that 1% to 2% sodium phosphate in 10% PVP causes a second glass transition to be observed in the low-temperature thermogram, indicating a phase separation in the freeze concentrate. Comparison of DSC thermograms of fast frozen solutions of sucrose with and without added sodium chloride shows that electrolyte-induced reduction of Tg' is not caused by a direct plasticizing effect of the electrolyte on the freeze concentrate. Measurement of unfrozen water content as a function of temperature by a pulsed nmr method shows that the most likely mechanism for electrolyte-induced changes in Tg' is by increasing the quantity of unfrozen water in the freeze concentrate, where the unfrozen water acts as a plasticizer and decreases Tg'. The correlation time (tau c) of water in the freeze concentrate is in the range of 10(-7) to 10(-8) seconds. The results underscore the importance of minimizing the amount of added salts to formulations intended for freeze drying. PMID- 7479567 TI - Process-induced crystallinity changes in albuterol sulfate and its effect on powder physical stability. AB - Pharmaceutical powders are often milled to achieve the optimum particle size. These size reduction processes can introduce dislocations and/or defects onto particle surfaces affecting the overall crystallinity of the powder. If enough energy is imparted, amorphous regions on the particle surfaces may be produced. These amorphous regions have the propensity to absorb significant quantities of water. In this study the effect of sorbed water on the physical characteristics of albuterol sulfate is investigated. Physical properties of this compound are studied in both micronized and unmicronized states using scanning electron microscopy, differential scanning calorimetry, powder x-ray diffraction, solution microcalorimetry, laser diffraction particle size analysis and water vapor sorption analysis. Subtle differences in crystallinity induced by air jet micronization are detected by several analytical methods. Amorphous to crystalline conversions are observed, the kinetics of which are found to be both temperature and relative humidity dependent. These experiments show the dynamic nature of micronized albuterol sulfate and aid in the determination of the actual physical state of this pharmaceutical powder. PMID- 7479569 TI - Drug release from semisolids: effect of membrane permeability on sensitivity to product parameters. PMID- 7479568 TI - Gastric pH influences the appearance of double peaks in the plasma concentration time profiles of cimetidine after oral administration in dogs. AB - The plasma concentration-time profiles of cimetidine often exhibit two peaks following oral administration of a single dose in the fasted state, while the concurrent administration of some antacids results in a lower extent as well as rate of absorption. In the present work, absorption of cimetidine after a single dose in the fasted state was studied as a function of gastric pH in male beagle dogs to determine whether gastric pH plays a role in the double peak phenomenon and/or can account for the decrease in bioavailability when antacids are coadministered. The extent of absorption of cimetidine was not influenced significantly by gastric pH, indicating that elevation of gastric pH is not the cause of decreases in the bioavailability of cimetidine when it is administered with antacids. Distinct double peaks or plateaux were noted in 8 of 10 plasma profiles when the gastric pH was 3 or below. Irregular absorption behavior was observed in 2 of 6 profiles in the pH range of 3 to 5, while single peaks were observed in all 10 profiles when the gastric pH was maintained at pH > or = 5. It was concluded that gastric pH is a major factor in the generation of cimetidine double peaks. Changes in gastric pH also resulted in changes in the apparent kinetics of absorption. Below pH 5, absorption mostly followed zero-order kinetics (9 of 16 profiles) or a more complex kinetic process involving at least two components to the absorption phase (5 of 16 profiles). At gastric pH > or = 5, however, absorption followed first order kinetics in 7 of 10 profiles. These differences in kinetics of absorption are postulated to arise from variations in gastric emptying as a function of pH and/or carryover effects of gastric pH into the upper intestine. PMID- 7479571 TI - Dose-dependent brain delivery of zidovudine through the use of a zidovudine chemical delivery system. PMID- 7479570 TI - Model features of a cardiac iontophoretic drug delivery implant. PMID- 7479572 TI - Effects of lead and mercury on histamine uptake by glial and endothelial cells. AB - The effects of lead and mercury on [3H]-histamine uptake by cultured astroglial and endothelial cells of rat brain were studied. Experimental data showed that both metal ions inhibited the uptake in both cell types of concentrations as low as 1-10 microM. The effects were consistent with non/competitive inhibitions. With either lead or mercury exposure, the inhibition of the uptake was greater in astroglial than in cerebral endothelial cells. Contrary to the above findings, 100 microM of mercuric chloride produced stimulation of histamine uptake and this stimulation was much more pronounced in cultured cerebral endothelial cells than in astroglial cells. Inhibition of [3H]-histamine uptake by lead acetate and mercuric chloride was considered to be association with a loss of the transmembrane Na+ and/or K+ gradient while stimulation of the uptake by high concentration of mercury might be related to a direct effect on histamine transporter. It is noteworthy, that cultured astroglial cells, derived from neonatal rat brain, are much more sensitive to the toxic effects of these heavy metal ions than cultured endothelial cells derived from the brain capillaries of the same species of animals. PMID- 7479573 TI - Effects of platelet-activating factor antagonists WEB 2086 and BN 50730 on digoxin-induced arrhythmias. AB - Effects of platelet-activating receptor antagonists WEB 2086 (1.0-30.0 mg.kg-1 intravenously) and BN 50730 (10.0 mg.kg-1 intravenously) alone or in combination with CGS 8515 (a specific 5-lipoxygenase inhibitor, 0.3 mg.kg-1 intravenously) and Dazmegrel (a thromboxane synthase inhibitor, 1.0 mg.kg-1.hr-1 intravenous infusion) on digoxin-induced arrhythmias were investigated in anaesthetised guinea-pigs. ECG, mean arterial blood pressure, heart rate and arrhythmias were recorded, starting 30 min. before digoxin administration and continuing for 60 min. afterwards. WEB 2086 (10.0 mg.kg-1 intravenously) reduced the mortality rate and arrhythmia score significantly compared to the control values. However, in combination with CGS 8515, it did not affect the mortality rate. BN 50730 (10.0 mg.kg-1) reduced the incidence of ventricular fibrillation and also arrhythmia score. BN 50730 in combination with Dazmegrel was reduced the arrhythmia score, incidence of ventricular fibrillation and mortality rate significantly, compared to control values. Digoxin-induced acute rise in mean arterial blood pressure was not affected by any of drug treatment except WEB 2086 (10.0 mg.kg-1) in combination with CGS 8515. Heart rate values did not differ between groups. However, pressure-rate index was reduced by WEB 2086 alone or in combination with CGS 8615. Results showed that although two different platelet-activating factor antagonists have different effects on the incidence of ventricular fibrillation and mortality, they improved the digoxin-induced arrhythmias when they were used either separately or in combination with CGS 8515 or Dazmegrel by implicating that platelet-activating factor has a role on digoxin-induced arrhythmias. PMID- 7479574 TI - Depletion of copper and manganese in brain after MPTP treatment of mice. AB - The mechanism of action of MPTP, a parkinsonism-inducing drug has been related to trace metals as a result of the observed potentiation of the neurotoxic action of the drug when diethyldithiocarbamate is concurrently administered. Diethyldithiocarbamate is a well-known chelator of trace metals, particularly copper. In the present study we analyzed the concentrations of copper and manganese in four brain regions of mice treated with neurotoxic doses of MPTP, in order to further substantiate the relationship between trace metals of MPTP induced neurotoxicity. Male Swiss albino mice were administered with MPTP (30 mg/kg) for either three or five days. Seven days after the last MPTP administration, they were sacrificed and the content of manganese and copper in the following regions was determined by graphite furnace atomic absorption spectrophotometry: cortex, cerebellum, midbrain and corpus striatum. Results indicate a significant depletion of manganese in corpus striatum (19.5% versus control) in the mice treated with MPTP for 5 days. Copper was also found to be decreased in corpus striatum (17.3% in mice treated for 3 days and 51.3% in mice treated for 5 days). Midbrain copper was depleted by 42.9% in the group of mice treated for 5 days with MPTP. Results indicate that MPTP induced a diminution of both copper and manganese in corpus striatum, suggesting that this alteration could be related to MPTP mechanism of action. PMID- 7479575 TI - Comparison of the binding activities of some drugs on alpha 2A, alpha 2B and alpha 2C-adrenoceptors and non-adrenergic imidazoline sites in the guinea pig. AB - Simultaneous computer modelling of control and guanfacine-masked [3H]-MK 912 saturation curves as well as guanfacine competition curves revealed that both alpha 2A- and alpha 2C-adrenoceptor subtypes were present in the guinea pig cerebral cortex. The Kd value of [3H]-MK 912 determined for the alpha 2A-subtype was 403 pM and for the alpha 2C-subtype 79.8 pM; the receptor sites showing capacities 172 and 19.5 fmol/mg protein, respectively. The Kds of guanfacine were 20 and 880 nM for the alpha 2A- and alpha 2C-adrenoceptor, respectively. In the guinea pig kidney [3H]-MK 912 bound to a single saturable site with Kd 8.34 nM and capacity 285 fmol/mg protein, the site showing pharmacological properties like an alpha 2B-adrenoceptor. Binding constants of 22 compounds for the three guinea pig alpha 2-adrenoceptor subtypes were determined by computer modelling competition curves using for the cerebral cortex a "3-curve assay", for the kidney an "1-curve assay", and using [3H]-MK 912 as labelled ligand. Of the tested drugs guanfacine and BRL 44408 were found to be clearly alpha 2A selective, Spiroxatrine, yohimbine, rauwolscine and WB 4101, as well as [3H]-MK 912 itself, were found to be alpha 2C-selective. The most selective compounds for alpha 2B-adrenoceptors, when compared to alpha 2A-adrenoceptors, were ARC 239 and prazosin. In the guinea pig kidney [3H]-p-aminoclonidine bound to alpha as well as to non-adrenergic imidazoline sites. The alpha 2-adrenoceptors could be completely blocked using 10 microM (-)-adrenaline without the non-adrenergic sites being affected. During these conditions the analysis of combined saturation and competition studies using labelled and unlabelled p-aminoclonidine with computer modelling revealed that the ligand labelled two different sites with Kds of 310 and 47,000 nM, respectively. Competition curves of 16 compounds for the non-adrenergic [3H]-p-aminoclonidine sites were shallow and resolved into two site fits. For the high affinity [3H]-p-aminoclonidine site the highest affinities were shown by 1-medetomidine, UK-14,304, guanabenz and detomidine; the Kds of these drugs ranging 26-72 nM. All drugs tested showed low but varying affinities for the low affinity [3H]-p-aminoclonidine site. These data indicated that the [3H]-p-aminoclonidine binding sites of the guinea pig kidney are grossly different from the [3H]-idazoxan binding I2-receptors previously demonstrated also to be present in the guinea pig kidney. PMID- 7479576 TI - 1 alpha,25-Dihydroxyvitamin D3 decreases scalding- and platelet-activating factor induced high vascular permeability and tissue oedema. AB - We have previously demonstrated that 1,25-dihydroxyvitamin D3 decreases endotoxin induced mortalities of rat and mouse from 80% and 70% to 14% and 10% (unpublished results). In the present study, pretreatment of 1,25-dihydroxyvitamin D3 decreased pad oedema and pulmonary vascular permeability induced by scalding and platelet-activating factor. 1,25-Dihydroxyvitamin D3 in a dose of 4 x 10(-9) mol/kg decreased pulmonary extravascular Evans blue concentration increased by platelet-activating factor from 6.32 +/- 0.63 ng/g to 2.57 +/- 0.28 ng/g (P < 0.01), and lung water/dry lung weight from 3.79 +/- 0.57 to 1.51 +/- 0.13 (P < 0.01). 1,25-Dihydroxyvitamin D3 of 4 x 10(-11) moles/kg and calcipotriol (a vitamin D analogue which has limited hypercalcaemic effect) of 4 x 10(-9) moles/kg had similar effects. Karyocyte counting of bronchoalveolar lavage fluid and the reduction of mean arterial pressure were also reduced to some extent. These results suggest that 1,25-dihydroxyvitamin D3 may be a new and effective drug to reduce vascular permeability and ameliorate oedema. PMID- 7479577 TI - Is there any cross-antagonism between mu-opioid and alpha 2-adrenergic receptors in the rat spinal cord? AB - A functional connection, evidenced by synergism and cross-tolerance, seems to exist between the mu-opioid and the alpha 2-adrenergic receptors in the superficial dorsal horn of the rat spinal cord. In this study possible cross antagonism between these systems was studied by intrathecal administration of morphine 2.5 micrograms or dexmedetomidine 1.25 micrograms 15 min. after subcutaneous injection of naloxone 1.0 mg/kg or atipamezole 3.0 mg/kg in the morphine group and 4.0 mg/kg or 1.0 mg/kg in the dexmedetomidine group, respectively. No cross-antagonism of the antinociceptive effect was seen either in the tail flick or in the hot plate test at 15, 30, 45 and 60 min. from the pretreatment. PMID- 7479578 TI - Effects of long-term antiepileptic therapy on the catabolism of testosterone. AB - The serum levels of testosterone, sex hormone binding globulin, and free testosterone index were measured in 51 epileptic men (age 18-45) in order to assess the possible effects of antiepileptic drugs on sexual dysfunction. An analytical gas chromatography-mass spectrometry method was developed to assess the urinary excretion of testosterone, epitestosterone, androsterone, etiocholanolone, 11-OH androsterone and 11-OH etiocholanolone and to evaluate if the catabolism of testosterone had been increased. Twenty normal healthy males of similar age, 18-45 years, served as control group. Patients receiving polytherapy (n = 34) or monotherapy with carbamazepine (n = 8) or phenytoin (n = 9) showed higher levels of sex hormone binding globulin and testosterone, and lower levels of free testosterone than did the controls (P < 0.03). Urinary excretion of the metabolites androsterone and 11-OH androsterone was significantly reduced (P < 0.02) in the polytherapy group, while the monotherapy group showed only significant differences (P < 0.02) in the elimination of 11-OH androsterone. Our results suggest that an induction of the hepatic synthesis of sex hormone binding globulin may be the mechanism by which the antiepileptic drugs lower the levels of free testosterone in serum. However, the reduced excretion of androsterone and the normal levels of etiocholanolone show that the antiepileptic drugs do not produce an increase in the main catabolism pathway of testosterone. PMID- 7479579 TI - Differences in brain 5-HT transporter dissociation rates among animal species. AB - The potential of using receptor-ligand dissociation rates as a model for investigating molecular changes in receptors was tested using the dissociation of [3H]citalopram, [3H]paroxetine and [3H]imipramine from the brain 5-HT transporter of four different species (mouse, rat, pig and man). Since the dissociation rates of each of the three ligands differed in most of the species investigated, receptor-ligand dissociation rate constants would seem to be a sensitive measure of receptor conformation. The model could be useful in the search of structural variation in receptors whether attributable to genetic factors or to posttranslational modification. PMID- 7479580 TI - Different beta-adrenergic receptor density in different rat skeletal muscle fibre types. AB - The effects of adrenaline on skeletal muscle differ between fibre types. The aim of the present study was to investigate the beta-adrenoceptor density, affinity and subtype in rat skeletal muscles with different fibre type composition. beta Adrenoceptors were determined in cryostat sections to avoid methodological problems with variable recovery, using the non-selective beta-adrenoceptor ligand [3H]CGP-12177 and beta 1- and beta 2-selective cold ligands CGP 20712A and ICI 118,551. In the presence of protease inhibitors [3H]CGP-12177 binding was stable, saturable, reversible, and displaceable. Scatchard analysis of binding saturation data was compatible with a single class of specific binding sites. Binding site density (Bmax) was higher (P < 0.02) in adult soleus (9.38 +/- 1.13 fmol x mg protein-1) than in adult extensor digitorum longus (4.74 +/- 0.39 fmol x mg protein-1), whereas the dissociation constants (Kd), 0.37 +/- 0.05 and 0.31 +/- 0.04 nM for soleus and extensor digitorum longus, respectively, were not significantly different. For young rats (5-6 weeks), Bmax was 11.21 +/- 0.33 and 5.45 +/- 0.11 fmol x mg protein-1 (P < 0.05), and Kd was 0.27 +/- 0.02 and 0.24 +/- 0.04 nM for soleus and epitrochlearis, respectively. These results correspond to a receptor density of 2 and 1 pmol x g w.wt.-1 in muscles containing.mainly type I and type II fibres, respectively. Displacement studies with CGP 20712A and ICI 118,551 were compatible with mainly beta 2-adrenoceptors, but 7-10% beta 1 adrenoceptors were present in both types of muscle.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7479581 TI - Mechanism of cisplatin ototoxicity: antioxidant system. AB - The dose and duration limiting toxic effects of cisplatin are ototoxicity and nephrotoxicity. While several studies have attempted to shed some light on the causes of nephrotoxicity, the reasons for ototoxicity induced by cisplatin are poorly understood. Therefore, this investigation was undertaken to delineate the potential mechanisms underlying cisplatin ototoxicity. The role of glutathione (GSH), oxidized glutathione (GSSG) and malondialdehyde levels, and antioxidant enzyme activities [superoxide dismutase, catalase, GSH peroxidase, and GSH reductase] were examined in cochlear toxicity following an acute dose of cisplatin. Male Wistar rats were treated with various doses of cisplatin. Pretreatment auditory brain stem evoked responses (ABR) were performed and then post-treatment ABRs and endocochlear potentials were also performed after three days. Acute cochlear toxicity (ototoxicity) was evidenced as elevated hearing thresholds and prolonged wave I latencies in response to various stimuli (clicks and tone bursts at 2, 8, 16 and 32 kHz) on ABRs. The endocochlear potentials were reduced (50% control) in cisplatin-treated rats as compared to control animals. The rats were sacrificed and cochleae isolated. The GSH, GSSG and malondialdehyde levels, and antioxidant enzyme activities were determined. Cisplatin ototoxicity correlated with a decrease in cochlear GSH [0.45 +/- 0.012 nmol/mg] after cisplatin administration compared to 0.95-012 nmol/mg in control cochleae (P < 0.05). Superoxide dismutase, catalase activities and malondialdehyde levels were significantly increased in the cochleae of cisplatin injected rats. Cochlear GSH peroxidase and GSH reductase activity significantly decreased after cisplatin administration.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7479582 TI - Cytochrome P4502E1 inhibition by propylene glycol prevents acetaminophen (paracetamol) hepatotoxicity in mice without cytochrome P4501A2 inhibition. AB - Acetaminophen hepatotoxicity is associated with its biotransformation to the reactive metabolite N-acetyl-p-benzoquinone imine that binds to protein. Two forms of cytochrome P450, CYP2E1 and CYP1A2, have been implicated as primarily responsible for the bioactivation. To determine the relative contributions of these P450's, overnight fasted male NMRI mice were pretreated with 10 ml of 50% v/w propylene glycol/kg or fluvoxamine (10 mg/kg) at -80 and -20 min. relative to acetaminophen dosing to inhibit CYP2E1 and CYP1A2, respectively. Mice were sacrificed at 0.5 or 4 hr after a hepatotoxic dose of acetaminophen (300 mg/kg). Propylene glycol or propylene glycol plus fluvoxamine, but not fluvoxamine alone protected against acetaminophen hepatotoxicity as indicated by abolished increase in serum alanine aminotransferase activity, less depletion of hepatic glutathione and lower liver:body weight ratios. Propylene glycol inhibited the activity of CYP2E1 as indicated by 84% reduction in the clearance of 3 mg/kg dose of chlorzoxazone, whereas fluvoxamine inhibited the activity of CYP1A2 as indicated by 40% reduction in the clearance of a 10 mg/kg dose of caffeine. For this animal model, the data are consistent with the notion that hepatoxicity is associated with bioactivation of acetaminophen by CYP2E1 but not by CYP1A2. PMID- 7479583 TI - Immunosuppression, macroencapsulation and ultraviolet-B irradiation as immunoprotection in porcine pancreatic islet xenotransplantation. AB - Membrane encapsulation or ultraviolet-B irradiation, with or without mild immunosuppressive treatment, was applied in order to prolong the survival of xenogeneic porcine foetal pancreatic grafts. Non-diabetic C57BL/6 mice were transplanted with porcine islet-like cell clusters, either membrane-encapsulated in the epididymal fat pad, or non-encapsulated under the kidney capsule. The animals were treated with daily subcutaneous injections of either cyclosporin A (12.5 mg/kg b.wt.), 15-deoxyspergualin (5.0 mg/kg b.wt.), ethyl (E)-6- (1,3 dihydro-4-hydroxy-6-methoxy-7-methyl-3-oxo-5-isobenzofuranyl) -4-methyl-4- hexenoate. (RS-61443) (70 mg/kg b.wt.) or with cyclophosphamide (70 mg/kg b.wt.) every second day. A fulminant mononuclear cell infiltration was observed 14 days after transplantation both around the subcapsular graft and outside the membranes in the saline treated control group. The membrane had pores of 0.45 micron and was designed to allow macromolecule transport but prevents cells from crossing. Therefore, xenoantigens can escape from the membrane implants and cause an immune reaction. A significantly weaker mononuclear cell infiltration was, however, seen when the membrane barrier was combined with 15-deoxyspergualin, cyclophosphamide or RS-61443 treatment but the morphology of the encapsulated ICC was not improved. The best subcapsular, non-encapsulated graft survival was obtained in animals treated with 15-deoxyspergualin or cyclophosphamide and the graft insulin content measurements confirmed the morphological data. There was no prolongation of islet-like cell cluster graft survival under the kidney capsule after ultraviolet-B irradiation alone (650 J/m2 for 90 sec.), and no synergistic effect was observed when ultraviolet-B irradiation was combined with 15-deoxyspergualin therapy (2.0 mg/kg b.wt.).(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7479584 TI - Developmental toxicity of inhaled N-methylpyrrolidone in the rat. PMID- 7479585 TI - Crown-rump length in aneuploid fetuses: implications for first-trimester biochemical screening for aneuploidies. AB - This study examined the effect of estimation of gestational age from the menstrual history compared with that from crown-rump length (CRL) measurement on the detection rate of screening for aneuploidies in the first trimester. Pregnancy-associated plasma protein A (PAPP-A) was assayed in blood collected prior to chorionic villus sampling in 356 women with unaffected pregnancies and 28 women with an aneuploid pregnancy. There were 14 Down's syndrome (DS) pregnancies. All pregnancies were dated from menstrual history and CRL measurement. The average CRL gestation in the aneuploid population was 2.5 days less than that derived from the LMP (95 per cent confidence interval (CI) for LMP CRL gestation: using the algorithm based on unaffected pregnancies 0-3.5 days; using the matched case-control approach 1-4.5 days). The average CRL gestation in the DS population was 2 days less but this did not reach statistical significance (95 per cent CI for LMP-CRL gestation: using the algorithm -1 to 4.5 days; using the matched case-control approach 0 to 5.5 days). The detection rate of aneuploidies in the first trimester using maternal serum PAPP-A was reduced by 7 per cent (and by 3 per cent for DS) for a 5 per cent false-positive rate when using CRL rather than LMP to date the pregnancy. This phenomenon is a consequence of an apparent reduction of gestational age when estimated by CRL in the aneuploid population. Further studies are required to evaluate whether CRL is an unbiased estimate of gestation for Down's syndrome pregnancies. PMID- 7479586 TI - Prenatal diagnosis of congenital diaphragmatic hernia: timing of visceral herniation and outcome. AB - Ultrasonographic prenatal diagnosis of congenital diaphragmatic hernia is well established, but the correlation of prenatal detection with clinical outcome remains unclear. We report our experience with 15 cases of prenatally diagnosed congenital diaphragmatic hernia. Seven fetuses were detected at 14-16 weeks' gestation; two with a normal sonographic study at 15 and 16 weeks' gestation showed visceral herniation at 21 and 23 weeks, respectively. In the remaining six cases, a diaphragmatic hernia was found at ultrasonography after 24 weeks' gestation, while previous sonographic studies had been normal. All seven fetuses in whom a diaphragmatic hernia was diagnosed before 16 weeks' gestation were aborted; four of them had severe malformations or karyotype abnormalities. The two neonates who were diagnosed at 21 and 23 weeks' gestation died after surgical repair. In contrast, all six infants whose visceral herniation was diagnosed after 24 weeks of gestation, and whose sonographic studies at 15-23 weeks had been normal, are alive and well after corrective surgery. The results of this series suggest that the timing of visceral herniation into the thoracic cavity is a major indicator of the prognosis of these fetuses and that herniation that occurs after 25 weeks of gestation carries a favourable clinical outcome. Normal sonographic studies during the first half of pregnancy do not exclude the subsequent development of congenital diaphragmatic hernia, raising questions about the advisability of repeat examinations at later stages of gestation. PMID- 7479587 TI - Fetal ductus venosus flow velocity waveforms and maternal serum AFP before and after first-trimester transabdominal chorionic villus sampling. AB - Doppler flow velocity waveform recording in the fetal ductus venosus and umbilical artery as well as maternal blood sampling for serum alpha-fetoprotein (MSAFP) was performed before and after transabdominal chorion villus sampling (TACVS) in 36 women of advanced maternal age (> or = 36 years). Gestational age ranged between 11 and 13 weeks. No chromosomal anomaly was detected. No statistically significant difference was observed in ductus venosus velocity parameters or in the umbilical artery pulsatility index (PI) before and after CVS in 35 women with a normal pregnancy outcome. One case resulted in fetal loss. Post-CVS median MSAFP levels at 12 weeks (25 kIU/l) and 13 weeks (35 kIU/l) were significantly higher than pre-CVS levels. In three cases, post-CVS MSAFP levels were higher than 600 kIU/l, correlating with feto-maternal transfusions of approximately 1.0-1.4 ml, i.e., of around 40 per cent of feto-placental blood volume. One of these cases displayed absence of fetal peripheral blood flow velocities and fetal bradycardia following TACVS, resulting in fetal loss 1 week later. The remaining two cases had a normal pregnancy outcome, but showed a more than 50 per cent reduction in ductus venosus velocity after TACVS, whereas umbilical artery PI remained unchanged. However, similar velocity changes were associated with only small feto-maternal transfusions. Umbilical artery PI values remained unchanged. PMID- 7479588 TI - A chromosome 21-specific cosmid cocktail for the detection of chromosome 21 aberrations in interphase nuclei. AB - Fluorescent in situ hybridization (FISH) with a 21q11-specific probe (CB21c1) consisting of three non-overlapping cosmids has been applied to interphase amniocytes of pregnancies at increased risk for fetal aneuploidy (N = 78) and to interphase lymphocytes, cultured and uncultured, of patients referred for Down syndrome (N = 19 and 28, respectively). In the uncultured amniocytes, six chromosome aberrations were detected: three cases of trisomy 21, a triploidy, a de novo 46,XX,t(21q21q), and a mosaic 46,XY/47,XY,+dic(21)(q11)/48,XY,+dic(21)(q11),+del(21)(q11). In 15 cultured and 20 uncultured blood samples, FISH correctly diagnosed trisomy 21 (full or mosaic) at the interphase level, which was confirmed in all cases by subsequent karyotyping. Because of specific and strong signals in interphase nuclei, CB21c1 appears to be a useful tool for the rapid detection of chromosome 21 abnormalities. PMID- 7479589 TI - Risk-based prenatal screening for trisomy 18 using alpha-fetoprotein, unconjugated oestriol and human chorionic gonadotropin. AB - Nine centres collaborated to examine the feasibility of a screening method for trisomy 18 that was based on assigning individual risk, using a combination of maternal age and measurements of alpha-fetoprotein (AFP), unconjugated oestriol (uE3), and human chorionic gonadotropin (hCG). Second-trimester measurements of these analytes were obtained from 94 trisomy 18 pregnancies. In the 89 pregnancies without an associated open defect, the median levels for AFP, uE3, and hCG were 0.65, 0.43 and 0.36 multiples of the unaffected population median, respectively. The strongest individual predictor of risk for trisomy 18 was uE3, followed by hCG, AFP, and maternal age, in that order. Using a method of individual risk estimation that is based on the three markers and maternal age, 60 per cent of pregnancies associated with trisomy 18 would be detected at a risk cut-off level of 1:100, with a false-positive rate of about 0.2 per cent. One in nine pregnancies identified as being at increased risk for trisomy 18 would be expected to have an affected pregnancy. This risk-based screening method is more efficient than an existing method that is based on fixed analyte cut-off levels. Even though the birth prevalence of trisomy 18 is low, prenatal screening can be justified when performed in conjunction with Down syndrome screening and when a high proportion of women offered amniocentesis have an affected fetus. PMID- 7479590 TI - Molecular prenatal diagnosis of 3-hydroxy-3-methylglutaryl CoA lyase deficiency. AB - We report the first molecular prenatal diagnosis of 3-hydroxy-3-methylglutaryl CoA lyase (HL) deficiency. The proband had a classic but severe presentation with hypoketotic hypoglycaemia and acidosis, secondary mental retardation, and epilepsy, and HL deficiency was documented in cultured fibroblasts. We found him to be homozygous for the frameshift mutation N46fs (+1), which yields a distinct pattern on single-strand conformation polymorphism (SSCP) analysis. In two subsequent pregnancies, molecular prenatal diagnosis was performed using SSCP. In the first, chorionic villus biopsy was normal. In the second pregnancy, amniocentesis revealed an affected fetus. In both pregnancies, the diagnosis was confirmed enzymatically. HL activity was less than 7 per cent of control values in amniocytes and fetal liver of the affected pregnancy. In the second pregnancy, amniotic fluid metabolite measurements by stable isotope dilution-selected ion monitoring mass spectrometry showed greater than 100-fold increases of 3-hydroxy 3-methylglutaric acid and of 3-methylglutaconic acid levels compared with controls. PMID- 7479591 TI - The effect of thermal instability of intact human chorionic gonadotropin (ihCG) on the application of its free beta-subunit (free beta hCG) as a serum marker in Down syndrome screening. AB - The kinetics of free beta hCG concentrations were measured in 30 maternal whole blood samples from second-trimester pregnancies during 72 h incubation at 3, 20, and 30 degrees C. Dissociation of intact hCG (ihCG) was undetectable at 3 degrees C and produced a more than 20 per cent increase of free beta hCG at 20 degrees C and a more than 100 per cent increase at 30 degrees C. hCG dissociation at 30 degrees C was not reduced by a protease inhibitor (sodium iodoacetate) and also occurred in purified hCG dissolved in a protease-free incubation medium. These results were reproduced under conditions of sample transport by post at different environmental temperatures. In conclusion, reliable free beta hCG assessment requires that the specimen be kept cool from vene puncture until assay or completely other transport strategies have to be considered. Evaluation of free beta hCG as an effective marker in prenatal Down syndrome screening must be reconsidered from this aspect. PMID- 7479592 TI - Second-trimester levels of maternal urinary gonadotropin peptide in Down syndrome pregnancy. AB - Urinary gonadotropin peptide (UGP; beta-core fragment), a major metabolite of human chorionic gonadotropin (hCG), was shown recently to be markedly elevated in Down syndrome pregnancy between 19 and 22 weeks of gestation. To confirm and extend this finding, we obtained maternal urine and matching maternal serum samples from 14 cases of Down syndrome and six other aneuploidies between 17 and 21 weeks of gestation. UGP was measured in all these samples and in 91 singleton control urines. Results were corrected for urinary creatinine level and expressed as multiples of the control median (MOM). hCG levels were assayed in all serum samples from the cases and compared with previously established reference values. The median UGP level in Down syndrome cases was 5.34 MOM (range 2.71-12.57); 88 per cent of the values were above the 95th centile of control levels after modelling. The median maternal serum hCG level for the same cases was 2.20 MOM (range 0.84-3.40); 36 per cent of the values were above the 95th centile. The level of UGP in every case including all other aneuploidies was higher than the comparable maternal serum hCG level. Elevated UGP measurements are strongly associated with fetal Down syndrome during the second trimester and could contribute to improved Down syndrome screening protocols that are more accessible and less expensive than are currently available. PMID- 7479593 TI - Urinary multiple marker screening for Down's syndrome. AB - We have examined the possibility of using multiple markers in maternal urine rather than serum in order to screen for Down's syndrome. Urine samples were available from 36 cases (24 Down's syndrome, five Edwards' syndrome, three Turner's syndrome, one Klinefelter's syndrome, one triploidy, one triple-X, one twin discordant for Down's syndrome) and 294 controls, including three twins. Three markers were tested: the beta-core fragment of human chorionic gonadotrophin (hCG), total oestrogen (tE) and the free alpha subunit of hCG. Levels were corrected for creatinine excretion and expressed as multiples of the gestation-specific median (MOM) level from the singleton controls. The median value for the singleton Down's syndrome cases was 6.02, 0.74, and 1.08 MOM for beta-core-hCG, tE, and alpha-hCG, respectively. The increases in beta-core-hCG and the reduction in tE levels were highly significant (P < 0.0001 and 0.005, respectively; Wilcoxon rank sum test) but the increase in free alpha-hCG was not (P = 0.40). On the basis of a mathematical model, the expected detection rate for a 5 per cent false-positive rate was 79.6 per cent for beta-core-hCG alone, which increased to 82.3 per cent when combined with tE. Aneuploidies other than Down's syndrome were characterized by low levels of tE and either low or high beta-core hCG. PMID- 7479594 TI - Prenatal diagnosis of metatropic dwarfism. AB - We present a case of prenatal diagnosis of severe metatropic dysplasia at 20 weeks' gestation. The characteristic prenatal features of this rare autosomal recessive chondrodysplasia appear to be significant dwarfism with an enlarged head and a narrow thorax associated with enlargement of the hands and feet, and the radiographic 'dumb-bell' appearance of the long bones. PMID- 7479595 TI - Apparent segregation of null alleles ascribed to deletions of the ornithine transcarbamylase gene in congenital hyperammonaemia. AB - Ornithine transcarbamylase (OTC) deficiency is an X-linked disorder of the urea cycle mapped to chromosome Xp21.1. Here, we show that apparent segregation of null alleles at the OTC locus and flanking polymorphic loci mimicked false maternity or false paternity in three affected families. Based on these observations, we suggest giving consideration to gene deletion when dealing with segregation of null alleles in OTC deficiency. PMID- 7479596 TI - Ultrasound diagnosis of the Pena Shokeir phenotype at 14 weeks of pregnancy. AB - This report describes the early prenatal diagnosis of the Pena Shokeir phenotype in an at-risk patient at 14 weeks' gestation. The diagnosis was based on an abnormal fetal movement profile, in association with an abnormal position of the fetal limbs. Pena Shokeir phenotype describes an inherited condition characterized by arthrogryposis and dysmorphic features as a result of fetal akinesia. It is a lethal abnormality and early diagnosis allows safer surgical methods of termination. PMID- 7479597 TI - Prenatal ultrasonographic diagnosis of congenital megalourethra. AB - Congenital megalourethra is a rare genital anomaly characterized by dilatation of the penile urethra without evidence of distal obstruction. Reports of the prenatal diagnosis of this condition in the literature are limited. We present a case of congenital megalourethra with obstructive uropathy from the posterior urethra diagnosed prenatally at 18 weeks of gestation. 'Prune-belly'-like features, colonic malrotation, and imperforate anus were also found on autopsy. PMID- 7479599 TI - A case of paternal uniparental disomy for chromosome 11. AB - We report a case of paternal uniparental disomy for chromosome 11 that presented as severe intrauterine growth retardation. Autopsy following intrauterine death also revealed aberrant intestinal rotation and hypospadias. Chromosome analysis of direct preparations from placental biopsy showed an abnormal 47,XY,+11 karyotype. Analysis of long-term cultures from the placenta revealed 46,XY/47,XY,+11 mosaicism. Fluorescence in situ hybridization (FISH) studies on interphase nuclei confirmed trisomy 11 in multiple placental sites but detected only disomic cells in fetal skin. Investigation using microsatellite polymorphisms demonstrated paternal isodisomy at loci D11S909, D11S956, and D11S488, and paternal heterodisomy at locus D11S928. PMID- 7479598 TI - Prenatal diagnosis of trisomy 8 mosaicism in CVS after abnormal ultrasound findings at 12 weeks. AB - We describe a case of trisomy 8 mosaicism in which fetal chromosome analysis was prompted by ultrasound abnormalities, i.e., hygroma colli and dilatation of the renal pelves. Chorionic villus sampling (CVS) was performed, with a false negative result on direct karyotype analysis, although cultured trophoblasts revealed trisomy 8 mosaicism. Fetal autopsy confirmed the abnormalities found on ultrasound examinations and fetal tissue examination showed different levels of trisomy 8 mosaicism. To our knowledge, this is the first prenatal diagnosis of trisomy 8 made on ultrasound findings. PMID- 7479600 TI - Early prenatal diagnosis of ventricular diverticulum complicated by serous pericardial effusion. AB - An unusual case of apical diverticulum complicated by serous pericardial effusion and diagnosed ultrasonographically at 13 weeks of gestation is described. A therapeutic abortion was induced at 14 weeks and the complete post-mortem examination did not show additional malformation. Cardiac diverticulum is a rare malformation that occurs as an isolated defect or as part of a complex midline thoraco-abdominal defect. Only two prenatally diagnosed cases have been previously reported in the literature and none with associated hydropericardium. PMID- 7479601 TI - Effect of gravidity and parity on the parameters used in serum screening for trisomy 21. PMID- 7479602 TI - [Morphological typing, evaluation of tumor dignity and prognosis and etiologic classification of adrenomedullary and adrenocortical neoplasias]. AB - The aim of morphological tumour diagnosis is to answer clinical questions on type, biological potential, prognosis and aetiology of individual neoplasms. The limitations and perspectives of different methods used in the diagnosis of adrenal tumours, ranging from histology to molecular genetic DNA analyses, are described. When surgical specimens from adrenal neoplasms cannot be typed on the basis of histology and/or with clinica data (e.g., endocrine symptoms and history) as adrenocortical tumours, phaeochromocytomas or metastases to the adrenal, immunohistological investigations with a panel of different antibodies are necessary. After identification of the tissue derivation of an individual adrenal tumour, its biological potential must be assessed. Among adrenocortical neoplasms, adenomas and carcinomas can be distinguished by evaluation of various histological parameters (including structural features and signs of invasion) according to defined algorithms. In addition, conventional histology (by estimation of mitotic activity) allows the discrimination of tumours with especially high malignant potential from other adrenocortical carcinomas. In contrast, among adrenomedullary tumours even the combined use of histological, immunohistological and DNA cytophotometric techniques only allows the definition of risk groups (benign versus suggestive of malignancy), while reliable recognition of an individual malignant phaeochromocytoma is so far impossible. The question as to whether a particular phaeochromocytoma represents a sporadic tumour or a neoplasm inherited as one feature of a defined syndrome cannot be answered with the above methods, but only by the application of molecular genetic techniques. PMID- 7479603 TI - [Placental site trophoblastic tumor. Morphology, differential diagnosis and prognosis]. AB - A placental-site trophoblastic tumor is a rare neoplasia that is derived from the cells of the intermediate trophoblast. Morphological, biochemical, and Doppler ultrasound findings are presented regarding differential diagnosis using material from three recent cases. Essentially, placental-site trophoblastic tumors can be diagnosed if infiltration of the myometrium is seen by a monomorphic trophoblastic proliferation that is not interrupted by decidual cells. Necrosis and hemorrhages are not features of placental-site trophoblastic tumors. However, there is a peculiar behavior towards the uterine vasculature as spiral arteries are dilated and transformed the same way as occurs at the site of physiological implantation of pregnancy. It appears that as a result of this phenomenon there is a characteristic finding in ultrasound. Examination of this type of tumor demonstrates cystic spaces that can be defined as blood vessels by their Doppler signal. In two of the three cases a hysterectomy was performed, and criteria for the assumption of malignant placental-site tumors are therefore presented. However, only the mitosis rate seems to possess some predictive value. PMID- 7479604 TI - [Hippel-Lindau syndrome and sporadic renal cell carcinomas. Pathogenesis, morphologic spectrum and molecular genetics]. AB - von Hippel-Lindau (VHL) disease is an autosomal dominant heritable disease that often occurs in association with various benign and malignant tumours. Clinically the disease is classified as VHL 1 (without phaeochromocytoma) and VHL 2 (with phaeochromocytoma). Genetically, VHL is caused by germline mutations in the VHL tumour suppressor gene. More than 100 germline mutations and rearrangements have been identified, but the biological function of a hypothetical VHL protein is not yet known. Genotype-phenotype correlations should aid in the understanding of this biological role. All VHL manifestations subsequently develop to VHL mutations, but some mutations may act in a tissue-specific manner. Whereas missense mutations cause tumour suppression to fail in adrenal cells, more severe structural mutations are usually necessary for tumour development in renal cells. As predicted by the tumour suppressor theory, the VHL gene also plays a critical part in the pathogenesis of sporadic non-VHL-associated tumours. In a large number of sporadic renal clear cell carcinomas, mutations and hypermethylation cause inactivation of the VHL gene. Together with allelic 3p loss, these constitute rate-limiting events in renal tumourigenesis. Insights into the molecular basis of phenotypic variability in VHL disease and the confirmation of the tumour-suppressor criteria in VHL and non-VHL sporadic tumours indicate an important role of the VHL gene in the development of these tumours. PMID- 7479605 TI - [Primary pulmonary B-cell lymphoma of the MALT type]. AB - The malignant B-cell lymphoma of the MALT-type shows significant differences from nodal lymphomas with respect to its biological, molecular genetic, and clinical properties. According to the proposal from the International Lymphoma Study Group for a Revised European-American Classification of Lymphoid Neoplasms, MALT-B-cell lymphomas have been defined as an extranodal subgroup of marginal zone B-cell lymphomas and may have lower and higher grade types. We have applied this classification to 24 cases of primary B-cell lymphomas of the lung. These tumors showed histopathologic features of low (n = 20) and/or high grade neoplasms (n = 4) recapitulating the structure of the acquired bronchus-associated lymphoid tissue (BALT). According to our experience, the diagnosis of BALT-type B-cell lymphoma, is both practically feasible and reproducible not only for surgical material but even for biopsy specimens. PMID- 7479606 TI - [Arterial wall texture after uncomplicated endarterectomy]. AB - Recanalisation of the carotid sinus is one of the most frequently performed vascular interventions; endarterectomy of the coronary arteries is an auxiliary measure accompanying aortocoronary bypass. Enucleation of the sclerotic plaque is followed by reconstruction of the intima and by rapid smoothing over of the "steps" along the side of the recanalised segment by proliferation of smooth muscle cells from the remaining media. The thinner the residual media, the thicker grows the neointima. Occasionally elastic lamellae reminiscent of a lamina elastica interna are formed close to the lumen. Moreover, in some cases renewed formation of the characteristic intimal spur (fibrotic ridge) is seen at the carotid sinus inlet, a sign of continued irregularity of blood flow postoperatively. In contrast, no focal lesions are generally observed in recanalised segments in the coronary arteries. New sclerotic pads that occur in the late phase bear a morphological resemblance to those in operated restenoses. The distribution of these pads is largely congruent with that of the sclerotic plaques in non-operated cervical arteries. Thus, even when recanalisation of the carotid sinus is uncomplicated, reparative processes are followed by the reformation of sclerotic pads, largely determined by haemodynamic factors. In the coronary arteries, concentric rather than focal sclerotic lesions predominate. PMID- 7479607 TI - [New possibilities for structural analysis of bone biopsies using microcomputer tomography (muCT)]. AB - The combined histological and microcomputed analysis of human iliac crest biopsies leads to major advances in our understanding of three-dimensional bone architecture. Microcomputed tomography avoids the time-consuming reconstruction and artifacts of serial sections. Furthermore, its high resolution allows the recording of structural differences as low as 10 microns. Thus, three-dimensional analysis in combination with histological evaluation of cellular dynamics facilitates earlier and easier recording of changes of cancellous bone. PMID- 7479608 TI - [Neurocysticercosis]. AB - A 54-year-old female patient with a 10-year history of ventriculoperitoneal shunt resulting from communicating hydrocephalus of undetermined aetiology is reported. Transient gait disturbances and cerebral infarction at the age of 46 did not lead to further insights into the nature of the disease. After many years with only occasional disturbances, a distinct organic brain syndrome developed. Thorough examination led to a tentative diagnosis of neurocysticercosis; this was based on the history, liquor diagnosis and cerebral microcalcifications in CT. Despite the initiation of specific therapy, the patient died of the sequelae of the disease. At autopsy, characteristic cicatricial residues of mainly basal leptomeningitis were found with collapsed parasitic cysts. Additional intracerebral mesenchymal glial reactions were less conspicuous. Residual ependymitis had caused aqueductal stenosis. Death was due to cachexia, bronchopneumonia and a lung abscess. The clinical course and morphology of neurocysticercosis are discussed. The disease has become rare in our country, but is globally the most important parasitic disease of the central nervous system. PMID- 7479610 TI - [Ectopic hamartomatous thymoma. Case report with special reference to differential diagnosis]. AB - We report the case of an ectopic hamartomatous thymoma in a 56-year-old male patient. The lesion arose subcutaneously in the supraclavicular region. Histologically, the well-circumscribed but unencapsulated tumour was composed of uniform fusiform tumour cells. In addition, mature fatty tissue, scattered T lymphocytes, and an epithelial and a myoepithelial tumour cell component were found. The epithelial differentiation of the spindle cell tumour component was confirmed immunohistochemically and by electron microscopy. Ectopic hamartomatous thymoma has to be distinguished from ectopic cervical thymoma, thymolipoma, ectopic salivary tissue, teratoma, peripheral nerve sheath tumours, malignant epithelial tumours with thymus-like differentiation, biphasic synovial sarcoma, and skin adnexal tumours. PMID- 7479609 TI - [Prolactin producing hypophyseal carcinoma. Case report of an extremely rare metastatic tumor]. AB - A 59-year-old male patient was transnasally operated on because of a pituitary adenoma with hypopituitarism. A second operation and X-ray therapy followed a half year later due to recurrent tumor. Both neoplasmas were classified as sparsely granulated prolactin cell adenomas. Immunohistochemical studies revealed strong immunoreactivity for prolactin and FSH in the tumor cells of both the pituitary adenoma and the recurrent tumor. Two years later the prolactin plasma levels were extremely elevated. A tumor in the liver was identified. Biopsy revealed a solid endocrine tumor containing prolactin by immunohistology. Due to structural and immunohistological similarities this tumor could be identified as a metastasis of the pituitary tumor. After 5 months of therapy the patient died from thrombembolism. Post-mortem studies confirmed the diagnosis of a metastasizing prolactin-secreting pituitary carcinoma. Only six similar cases have been reported in the literature. Our case report confirms the experience with 35 definite pituitary carcinomas reparted in the current literature: malignant pituitary tumors develop after pituitary surgery and can be identified not from the pituitary tumor, but only from its metastases. PMID- 7479611 TI - [Detection of atypical mycobacterial endophthalmitis by PCR]. AB - We report a case of a 60 year old male who presented with a therapy resistant endophthalmitis which led to an evisceratio bulbi. Histologically, a granulomatous inflammation was present, in which acid-fast bacilli could not be detected. Molecular genetic examination by means of polymerase-chain-reaction technique (PCR) of the routinely processed tissue material revealed the diagnosis of an atypical mycobacterial infection, because DNA specific for atypical mycobacteria was detected. This example shows that PCR technique provides a useful tool in the diagnosis of mycobacterial infections as an adjunct to surgical pathology. PMID- 7479612 TI - Transplacental clearances of inert hydrophilic tracers in rabbits of 18 days gestation. AB - Transplacental clearances were measured for radiolabelled Cl-, SO4(2-), mannitol, sucrose, raffinose, PEG-900 and inulin. Maternal placental blood flows were measured with radiolabelled microspheres. At 18 days of gestation (term 31 days), the fetuses and extra-fetal fluids were growing at 55 and 32 per cent per day, accounting for a net transplacental filtration rate of 14.2 nl/sec per gram placental weight. Pore theory and a least squares fit of the Patlak equation yielded an equivalent pore radius of 1.75 nm. It was demonstrated that the clearance of the largest tracer, inulin, was 30 times higher than it would have been in the absence of net filtration. Comparison with literature data showed that there was a small increase in placental permeability per gram placenta between 14 and 18 days of gestation but that the increase between 18 and 28 days of gestation was about 14-fold for Cl- and 300-fold for inulin. There was no evidence for a decreasing equivalent pore radius in the course of gestation from 14-18 days. PMID- 7479613 TI - Characteristics of trophoblast cells migrating from first trimester chorionic villus explants and propagated in culture. AB - We developed a method of propagating pure first trimester human trophoblast cells growing out of primary explants of mechanically derived chorionic villus fragments (Yagel et al, 1989; Graham et al, 1992). We have now extensively characterized these cells during their initial outgrowth and in long-term culture, employing a variety of markers and techniques as outlined below. By double label immunofluorescence using epithelial (cytokeratin) and mesenchymal (vimentin) cell markers, we identified the chorionic villus migrant cell populations as pure trophoblast (39 per cent of outgrowths) or a mixture of trophoblast and fibroblast (61 per cent). Further phenotyping of the pure trophoblast outgrowths by double label immunostaining using anti-cytokeratin antibody and a panel of other primary antisera revealed that these cells exhibit a variety of markers characteristic of extravillous invasive trophoblast cells in situ: insulin-like growth factor (IGF)-II, NDOG-5, proliferating cell nuclear antigen (PCNA), human leucocyte antigen framework antigen (W6/32) and a distinct set of integrins including alpha 1, alpha 3, alpha 5, alpha v and beta 1 subunits and alpha v beta 3/beta 5 vitonectin receptor. They were negative for alpha 6 and beta 4 integrin subunits. Immunogold electron microscopy of explants grown on type IV collagen gel revealed the production of conventional and oncofetal types of fibronectin by mononucleate trophoblast cells and human placental lactogen by multinucleate cells. Immunolabelling, flow cytometry and immunoprecipitation revealed that this phenotypic profile was retained with complete fidelity in the long-term culture; thus, trophoblasts migrating out of first trimester chorionic villus explants and their propagated progeny belong to the invasive extravillous trophoblast of the placenta. PMID- 7479614 TI - Increased immunohistochemical expression of neutral metalloendopeptidase (enkephalinase; EC 3.4.24.11) in villi of the human placenta with pre-eclampsia. AB - The purpose of this study was to identify the presence of placental neutral metalloendopeptidase (NEP; enkephalinase; EC 3.4.24.11) in human normotensive and pre-eclamptic pregnancy. The localization of NEP in placentae from normotensive, chronic hypertensive and pre-eclamptic pregnancies was carried out on fresh frozen tissues by using a monoclonal primary antibody developed against human common acute lymphoblastic leukaemia antigen (CD10) together with the avidin biotin-peroxidase method. In placentae from normotensive, chronic hypertensive and superimposed pre-eclamptic pregnancies, intense staining was found in the extravillous trophoblast, and also in fibroblasts of the chorionic plate and stem villi. Light to moderate staining was noted in the villous-associated trophoblast and in some cells from the villous core. In cases of pre-eclampsia, very intense staining was detected not only on the surface, but also in the cytoplasm of the villous-associated trophoblast. The increased expression of placental NEP in pre eclampsia suggests that this enzyme may be involved in the regulation of the local concentration of circulating biologically active peptides at the fetomaternal interface, and thus could be implicated in the pathophysiological changes of this syndrome. PMID- 7479615 TI - Hypertension in pregnancy: changes in activin A maternal serum concentration. AB - Human placenta is the major source of activin A in maternal circulation. The aim of the present study was to evaluate maternal activin A serum concentration in pregnant women with chronic hypertension (n = 14), pregnancy-induced hypertension (n = 10) or pre-eclampsia (n = 16). In the group of pregnant women with chronic hypertension and of healthy pregnant women (n = 10) activin A was measured in samples collected longitudinally throughout gestation. Using a specific two-site enzyme-linked immunosorbent assay, it has been possible to measure maternal serum activin A concentration. In addition, the effect of recombinant human activin A administration on mean arterial pressure and heart rate in female rats have been also investigated. Mean +/- SEM of maternal serum activin A concentration in pre eclamptic women (57.4 +/- 28.3 ng/ml), was significantly higher than in women with pregnancy-induced hypertension (14.8 +/- 10.5 ng/ml), chronic hypertension (10.3 +/- 5.4 ng/ml) or healthy control women (9.2 +/- 9.4 ng/ml) (P < 0.01). Serum activin A levels evaluated 2 weeks after anti-hypertensive treatment were not significantly different in pre-eclamptic women. Moreover, when exogenous recombinant human activin A was administered in female rats arterial pressure or frequency of heart rate did not change. The present study showed that maternal serum activin A concentration is abnormally high in patients with pre-eclampsia. Thus, since the patients with chronic hypertension or pregnancy-induced hypertension have activin A concentration in the normal range of values, activin A may be a prognostic marker of hypertension in pregnancy. PMID- 7479616 TI - Placental isoferritin levels during first trimester of normal and complete molar gestations. AB - Serum placental isoferritin levels (PLF) levels were measured in 33 patients admitted for routine scanning of a first trimester normal singleton pregnancy and six patients who were hospitalized for uterine evacuation of a complete molar gestation. Venous blood was obtained upon admission and before curettage, when necessary. Serum was separated into glass tubes, immediately frozen and stored at -20 degrees C until analysed. The mean serum PLF levels were 18.1 (+/- 14) U/ml and 5.5 (+/- 2) U/ml for normal and molar gestations, respectively, with a significant difference between the two groups (P = 0.001). Sixty-seven per cent of normal pregnancies had serum PLF levels > or = 10 U/ml, whereas none of the molar gestation group reached this threshold level. Furthermore, the molar gestation group's low serum PLF levels remained unchanged throughout the entire follow-up period and until their beta-human chorionic gonadotrophin levels were undetectable. Unlike normal pregnancies, the molar trophoblast does not seem to secrete or synthesize PLF, suggesting that the complete molar placenta has different protein-producing capabilities when compared with those found in normal pregnancies. Further studies, including serum PLF among other categories of gestational trophoblastic neoplasms, are recommended before this data can be integrated into routine clinical work. PMID- 7479617 TI - The study of mitochondrial gene modifications in human placenta. AB - A quantitative study on the effects of placental senescence on mitochondrial DNA (mtDNA) was carried out by measuring mitochondrial gene mutation, levels of mRNA for the cytochrome c oxidase subunit I (COI) and of mtDNA in normal placental tissues at different stages. Deleted mtDNA, the expression of COI in mitochondria and the amounts of mtDNA per cell were examined by the polymerase chain reaction, northern blot analysis and southern blot analysis, respectively. No accumulation of mutant mtDNA with a 4977 base pair (bp) deletion was detected in the normal placenta during pregnancy. There appeared to be a gradual increase of COI mRNA as pregnancy progressed, while the ratio of mtDNA to total DNA in human placenta tended to decrease with gestation. These results indicate that placental ageing is not associated with the accumulation of mtDNA mutation with the 4977 bp deletion or markedly reduced expression of the mitochondrial genome. PMID- 7479618 TI - Birthweight and sodium/proton exchanger activity in the term human placenta. PMID- 7479619 TI - Conference report: placental growth and function. PMID- 7479620 TI - Exercise in middle-aged adults: self-efficacy and self-presentational outcomes. AB - BACKGROUND: Whereas self-efficacy expectations have been identified as important determinants of exercise participation patterns, little empirical work that examines efficacy expectations as outcomes of exercise participation or their theoretical relationship to other psychological outcomes associated with exercise has been conducted. In the context of middle-aged males and females, the present study attempted to integrate social cognitive and impression management perspectives with respect to anxiety associated with exercise. METHODS: Formerly sedentary subjects participated in a 5-month exercise program with assessments of physique anxiety, efficacy, outcome expectations, and anthropometric variables prior to and following the program. RESULTS: Both acute bouts and long-term participation in exercise resulted in significant increases in self-efficacy. In turn, these changes in efficacy and initial positive outcome expectations were significant predictors of reductions in physique anxiety, even when controlling for the influence of gender and reductions in body fat, weight, and circumferences. CONCLUSIONS: The findings are discussed in terms of the implications for structure and content of exercise environments and the utility of the proposed theoretical integration. Strategies for enhancing beliefs regarding health and fitness outcomes associated with exercise rather than appearance outcomes may be required to maximize reductions in negative body image. PMID- 7479621 TI - Potential sources of multiple mutations in human cancers. AB - Increasing evidence indicates that most human cancers contain multiple chromosomal alterations. These aberrations are the result of mutations produced not only during the initiation of cancer but also during tumor progression. Since the rates of spontaneous mutations exhibited by normal human cells cannot account for the large numbers of mutations routinely reported in human cancers, we argued that cancer cells are genetically unstable; i.e., they express a mutator phenotype. In this review, we consider potential endogenous sources of these mutations and the recent evidence demonstrating that multiple mutations are present in human cancers. These studies, which connect mismatch repair, genomic instability, and cancer, support the mutator phenotype hypothesis. We conclude that, if multiple mutations are necessary for the progression of cancer, then agents designed to delay their accumulation could significantly reduce cancer deaths. PMID- 7479623 TI - Commercial availability of alcohol to young people: results of alcohol purchase attempts. AB - OBJECTIVES: Commercial availability is an important contributor to alcohol use by young people. The purpose of this study was to test the purchasability of alcohol by youth who appeared to be under the legal age of sale in all types of businesses licensed to sell alcohol in upper Midwest communities. METHODS: Female buyers who were age 21 but whose apparent age as rated by a panel averaged 19 years attempted to purchase beer without age identification at all off-sale outlets (n = 395) and a random sample of on-sale outlets (n = 492) in 24 communities in Minnesota and Wisconsin. Two purchase attempts were made at each outlet, with the buyer randomly assigned to outlet and community. Characteristics of the outlets, clientele, signs, interaction with the sellers or servers, and purchase success were recorded. RESULTS: The buyers were successful in purchasing beer without age identification in 50% of attempts in on-sale outlets and 52% of off-sale attempts. Almost 75% of the outlets sold beer to the buyers at least once in two attempts. In off-sale businesses buyers were more successful if the salespersons were male and if the businesses were located in residential areas or malls. On-sale purchase attempts were more likely to be successful if the server appeared to be younger than 30 years, if the business was a restaurant as opposed to a bar alone, and if signs warning against sales to minors were located at the checkouts. CONCLUSIONS: The results of this study confirm and extend previous reports of the ability of youth to purchase alcohol and support the conclusion that commercial availability of alcohol to youth is widespread. PMID- 7479622 TI - Skin cancer risk and sun protection learning by helpers of patients with nonmelanoma skin cancer. AB - BACKGROUND: Knowledge-based skin cancer risk and prevention educational interventions by physicians and nurses were directed to subjects who had a nonmelanoma skin cancer. These high-risk patients asked relatives or friends to assist with postoperative care rendered after surgical removal of the skin cancer. The patient's experience with the nonmelanoma skin cancer was expected to raise the awareness of the helper. The study examined whether the patient became a source of information, risk assessment, and skills training for his or her helper. METHODS: Sequential patients between 30 and 60 years of age and their designated helpers completed a self-report questionnaire prior to the intervention and 1 year after the intervention. The questionnaire examined: (a) knowledge of skin cancer and sun protection; (b) individual susceptibility as determined by ease of sunburning or tanning; (c) attitudes about self-esteem, sun exposure, and health locus of control; (d) intentions to use sun protection; and (e) behaviors of sun protection used. RESULTS: Over the course of 1 year, 200 pairs were entered into the study. Both patients and helpers demonstrated an increase in knowledge after the educational intervention with the patients. There was a change in the self-reported intention to use sun protection and the behaviors of sun protection used in both patients and helpers after the intervention; however, there was no attitudinal change demonstrated in pre- and post-tests for either patients or helpers. Among both patients and helpers, gender-specific differences in attitudes, intentions, and behaviors existed. Women expressed a greater likelihood of taking precautions, including the use of sunblock; however, men reported the protective strategy of wearing a hat more than women did. Men valued a tan more than women and had greater outdoor exposure, which they restricted after the intervention. Women helpers ceased using indoor tanning devices after the intervention. Behavioral change in use of sun protection measures, including protective clothing or sunblock use and decrease in hours of outdoor sun exposure or use of indoor tanning devices, was dependent upon the patient or helper's own reported susceptibility as determined by his or her history of poor tanning and ease of sunburning. CONCLUSION: Patients transferred knowledge to their helpers. Intention to change behavior and behavioral change were strongly correlated with the individual's reported susceptibility to easy sunburning and poor tanning. Despite a lack of change in attitudes, changes in both intention to change behavior and behavior itself occurred in those who perceived themselves to be at risk. While the attitudes of participants in this study reflected popular beliefs, targeted education of high risk adults with a nonmelanoma skin cancer caused changes in sun protection behaviors in both the patients and their helpers. PMID- 7479624 TI - Recruitment to a university alcohol program: evaluation of social marketing theory and stepped approach model. AB - BACKGROUND: This study was a first initiative to evaluate the application of social marketing theory (SMT) to increase attendance at an alcohol abuse education program for university residence hall students and to ascertain whether aggressive recruitment strategies are necessary as part of the stepped approach model (SAM) of service delivery. METHOD: SMT and public health strategies that include focus groups, in-depth interviews, and intercept interviews were used to develop recruitment materials in a Test Hall. These new recruitment materials were introduced to the residents in the Treatment Hall (N = 727) and were compared to the Usual Care, Control Hall (N = 706) which received the recruitment materials normally provided to residents as well as to three Historical Halls separately and combined which had used the Usual Care recruitment materials in the past. RESULTS: The Treatment Hall percentage attendance was significantly superior (0.001 < p < 0.05) in all comparisons. The percentage attendance did not differ significantly from marketing literature expectations. The projections for campus-wide attendance for residence hall students were between 207 and 243 participants and for nationwide attendance, 36,900 +/- 8,185. CONCLUSIONS: The results suggest that the SMT and public health methods used are helpful in developing recruitment strategies and are an important initial step of the SAM and that a "minimal intervention" recruitment strategy is a cost-effective approach that can have a dramatic impact. PMID- 7479626 TI - Can we target smoking groups more effectively? A study of male and female heavy smokers. AB - BACKGROUND: In recent years a number of studies have examined subgroups within the smoking population. These studies have provided information which facilitate improved targeting of programs and interventions to help smokers quit. Despite this there have been few population studies which have described male and female heavy smokers and compared their characteristics. METHODS: We used representative population data on 789 smokers to examine differences between light and heavy smokers of each gender and between male and female heavy smokers directly. RESULTS: Of those who smoked, 35% of males and 24% of females were classified as heavy smokers and represent a large target group within the smoking population. Whereas a light smoker was characterized as smoking a median of 12.5 cigarettes per day, the heavy smoker smoked a median of 32 cigarettes per day. Univariate comparison of heavy smokers and light smokers showed a number of statistically significant differences according to demographics, in smoking behaviors and knowledge, in beliefs about smoking, in smoking characteristics, and in other health indicators. Because of interactions between gender and work/leisure day smoking variation, separate logistic regression models based on gender were fitted to the data. Again, compared with light smokers, several significant differences emerged for both heavy female and heavy male smokers under the headings mentioned above. Further logistic regression analyses showed differences between male and female heavy smokers when compared directly. CONCLUSIONS: The smoking population of South Australia is not homogeneous and this segmentation study has shown the need for varying approaches to different segments when developing intervention programs. PMID- 7479625 TI - Effects of treatment on cardiovascular risk among smokeless tobacco users. AB - BACKGROUND: Studies show sustained levels of nicotine among young males using smokeless tobacco, causing concern for subsequent cardiovascular risk. Also, there is little information on effects of nicotine replacement on cardiovascular risk in cessation programs. This study investigates the effects of nicotine gum replacement in smokeless tobacco cessation on cardiovascular risk factors. METHODS: Smokeless tobacco users, ages 18-65, were randomly assigned in a double blind fashion to 2-mg nicotine or placebo gum. At baseline, Week 4, and Week 8, dependent measurements, heart rate, blood pressure, and weight were recorded, and fasting lipoprotein profiles were drawn. RESULTS: This paper focuses on the smokeless tobacco users who refrained from use during the study period (N = 56). The nicotine gum group weighed less (P = 0.033) than the placebo group throughout the study and weight increased at a significant rate between Weeks 4 and 8 for both groups as gum decreased. Triglycerides were higher for the nicotine gum group than the placebo group (P = 0.031), with triglycerides decreasing between Weeks 4 and 8, with a similar effect seen among nonabstinent smokeless tobacco users. There was no dose, time, or dose by time effect for the other dependent measures. CONCLUSIONS: Among smokeless tobacco users who were abstinent, weight increased, with subjects on nicotine gum weighing less throughout the study. The lipoprotein profile, heart rate, and blood pressure did not improve over time, contrary to smokers in whom HDL increases and heart rate decreases with cessation. This could relate to different routes of administration, pharmacokinetics, or by-products of tobacco smoking being absent in smokeless tobacco. In addition, nicotine gum appeared to have neither an adverse nor a positive effect on heart rate, blood pressure, LDL, HDL, or total cholesterol. PMID- 7479627 TI - Medication cost savings associated with weight loss for obese non-insulin dependent diabetic men and women. AB - BACKGROUND: The cost of diabetes mellitus can be defined in both medical and economic terms. Although there is an abundance of literature concerning the medical management of diabetes, few studies have assessed the financial impact of its treatment. This study examined the savings in prescription costs associated with a weight reduction program for obese men and women with non-insulin dependent diabetes mellitus (NIDDM). RESEARCH DESIGN AND METHODS: Forty subjects ages 40-70 years who had body mass indexes of 30-40 kg/m2 and NIDDM of more than 1 year duration were assigned to one of two 800-kcal weight-loss programs for 12 weeks. A cost analysis was done on the 32 subjects who were taking anti hypertensive and/or anti-diabetes medications. A list of medications and monthly amounts was obtained at the start, upon completion, and 1 year following completion of the diet. The average out-of-pocket cost for a month's supply of each prescription was calculated by polling 16 retail pharmacies in Lexington, Kentucky. RESULTS: Subjects lost an average of 15.3 kg (33.7 lb) over the 12 weeks. At 1-year follow-up, subjects maintained a mean 9.0-kg (19.8 lb) weight loss. The average monthly prediet out-of-pocket cost for anti-hypertensive and anti-diabetes medications and supplies was $63.30 per subject. Following completion of the diet, this cost per month decreased to $20.40 and at 1-year follow-up the average monthly cost per subject was $32.40. The estimated average savings in prescription costs per subject over the year was $442.80. CONCLUSIONS: Significant short- and long-term savings in prescription costs were obtained following a 12-week hypocaloric weight reduction program for obese individuals with NIDDM. PMID- 7479628 TI - AIDS and cancer registry linkage: measurement and enhancement of registry completeness. The National AIDS/Cancer Match Study Group. AB - BACKGROUND AND METHODS: To measure AIDS registry completeness for Kaposi's sarcoma (KS) and cancer registry completeness for KS and non-Hodgkin's lymphoma (NHL), we linked AIDS and cancer registries at selected health departments. RESULTS: We found 10,350 people with KS: 1,935 reported only to the AIDS registry, 1,428 reported only to the cancer registry, and 6,987 reported to both. Ninety-three cases of non-HIV-associated KS were expected. For KS, AIDS registry completeness was 84% [6,987/(6,987 + 1,428 - 93)] and cancer registry completeness was 78% [6,987/(6,987 + 1,935)]. Cancer registry completeness for AIDS-related NHL was 76%. CONCLUSION: If this linking were conducted nationally, about 5,700 additional cases of AIDS and 10,000 additional cases of AIDS-related cancers would be recorded. PMID- 7479629 TI - Lifestyle intervention: results of the Treatment of Mild Hypertension Study (TOMHS). AB - BACKGROUND: Questions regarding the efficacy of nonpharmacologic approaches for the treatment of stage 1 hypertension were addressed as part of the Treatment of Mild Hypertension Study (TOMHS), a 4-year, randomized clinical trial (N = 902). This report describes the lifestyle intervention program used in TOMHS, presents data on the lifestyle changes observed, and focuses on the effect of weight loss on blood pressure and blood lipid levels. METHODS: Participants were randomly assigned to receive either placebo or one of five different antihypertensive medications. All took part in a lifestyle intervention program to reduce weight and sodium and alcohol intake and to increase physical activity. RESULTS: Substantial changes from baseline levels were achieved for all lifestyle intervention variables. Mean weight change was -10.5 lb (-5.6%) at 1 year, -8.5 lb (-4.5%) at 2 years, -7.4 lb (-4.0%) at 3 years, and -5.7 lb (-3.0%) at 4 years. At 4 years, 70% of participants remained below baseline weight and 34% maintained a weight loss of 10 lb or greater. Mean change in urinary sodium excretion was -12.5 mmol/8 hr (-23%) at 1 year, -10.7 mmol/8 hr (-20%) at 2 years, -8.4 mmol/8 hr (-16%) at 3 years, and -4.6 mmol/8 hr (-9%) at 4 years. Alcohol intake declined by 1.6 drinks/week among drinkers at 4 years. Reported leisure physical activity increased by 86% at 1 year and remained 50% above baseline at 4 years. Beneficial changes in blood pressure and serum lipids were associated with these changes. CONCLUSIONS: These results support a role for lifestyle interventions as the initial treatment for stage 1 hypertension and demonstrate that such interventions can be successfully implemented in the clinical setting. PMID- 7479630 TI - Smokers who fail to identify smoking cessation as a health priority. AB - BACKGROUND: Smokers who identified smoking cessation as the primary activity they could undertake to improve their health were compared with smokers who did not identify smoking cessation. Examined factors included personal characteristics, health status, smoking attitudes and behaviors, and social factors. METHODS: Subjects were 225 smokers who participated in a telephone survey of 853 randomly selected adults. RESULTS: Fifty-six percent of smokers did not identify smoking cessation as a priority. Nonidentifiers were more likely than identifiers to have minimal personal income, no paid employment, poor or fair health, dissatisfaction with their overall physical condition, less energy than others, little perceived control over their health, smoked fewer cigarettes per day, and reported no previous quit attempts. They were less likely to agree that they lacked the willpower to quit or that smoking helped them deal with stress. CONCLUSIONS: Smokers who fail to identify smoking cessation as a priority strategy for health improvement are unlikely to be exposed to workplace and social censures and should be targeted by health professionals. PMID- 7479631 TI - Coffee intake and risk of hip fracture in women in northern Italy. AB - BACKGROUND: Information on a possible relation between coffee, caffeine, and other methylxanthine-containing beverages and hip fracture is relatively scanty and controversial. We present here the results of a case-control study conducted in northern Italy. METHODS: A total of 279 cases of hip fracture and 1,061 controls in hospital for acute, nonneoplastic nontraumatic, non-hormone-related diseases were interviewed during their hospital stay. Odds ratios (OR) and their 95% confidence intervals (CI) according to consumption of coffee and other methylxanthine-containing beverages were derived from multiple logistic regression equations including terms for age, education, body mass index, smoking status, alcohol drinking, calcium intake, menopausal status, and estrogen replacement therapy. RESULTS: Compared with nondrinkers, the OR was 1.2 (95% CI, 0.8 to 1.7) for coffee drinkers. No association emerged with number of cups/day (OR = 1.2 for 1 cup/day, 1.0 for 2, 1.4 for 3 and 4, and 1.2 for 5 or more cups/day) or with duration of coffee intake (OR = 1.0 for less than 30 years and OR = 1.1 for more than 30 years). Similarly, no statistically significant association was observed with decaffeinated coffee (OR = 1.3), tea (OR = 1.3), or cola intake (OR = 0.6). OR for coffee drinking were computed across strata of age, menopausal status, education, smoking status, total alcohol drinking, and calcium intake. CONCLUSIONS: The present study found no association between hip fractures among women and consumption of regular or decaffeinated coffee, tea, and cola. PMID- 7479632 TI - Distribution of smokers by stage in three representative samples. AB - OBJECTIVES: A key variable for the design of individual and public health interventions for smoking cessation is Stage of Change, a variable which employs past behavior and behavioral intention to characterize an individual's readiness to change. Reactively recruited samples distort estimates of the stage distribution in the population because such samples attract a disproportionate number of late-stage participants. Three representative samples are described which provide accurate estimates of the stage distribution in the population. These samples are of adequate size to permit within-sample comparisons with respect to sex, age, Hispanic or non-Hispanic origin, race, and education level. The implications of using stage distribution as a tool for planning intervention is discussed. METHOD: The first sample of 4,144 smokers was from the state of Rhode Island and involved a random-digit-dial survey. The second sample of 9,534 smokers was from the state of California and involved a stratified random-digit dial survey. The third sample of 4,785 smokers was from a total of 114 worksites located in four different geographic locations. RESULTS: The stage distributions were approximately identical across the three samples, with approximately 40% of the sample in Precontemplation, 40% in Contemplation, and 20% in Preparation. The stage distribution was generally stable across age groups with the exception of the 65 years and older group. Education level did affect the stage distribution with the proportion of the sample in Precontemplation decreasing as education level increased. In all three samples, minor differences in stage distribution were related to Hispanic origin and race, but the pattern was not consistent across the samples. CONCLUSIONS: The pattern of stage distribution has important implications for the design of interventions. Existing interventions are most appropriate for the Preparation stage, but the majority of the three samples were in the first two stages, resulting in a likely mismatch between the smoker and the intervention. The stability of distribution across age suggests that interventions that are appropriately matched to stage can be applied across all age groups. The differences found with respect to education, Hispanic origin, and race can serve as a guide to the tailoring of intervention materials. PMID- 7479633 TI - Predictors of and satisfaction with bilateral prophylactic mastectomy. AB - BACKGROUND: Women with a first-degree relative with breast cancer are at increased risk of developing this disease. The optimal medical management of these women is unclear, with options including close breast cancer screening, bilateral prophylactic mastectomy, or participation in chemoprevention trials. Among women who undergo prophylactic bilateral mastectomy, very little is known about satisfaction with this surgery. Also, we know very little about variables related to prophylactic mastectomy decision making. METHODS: Participants were women at increased risk of breast cancer due to family history. These women were categorized by self-report as not interested in prophylactic mastectomy (n = 58), interested but deciding against surgery (n = 92), or subsequently having a bilateral prophylactic mastectomy (n = 14). Information on screening practices, risk perception, level of depression, and cancer-related worry was collected. Women completing prophylactic mastectomy reported on their satisfaction with the surgery and breast reconstruction. RESULTS: Women selecting surgery reported more breast cancer worry. The group expressing no interest in surgery reported fewer biopsies and lower risk estimates. Women completing surgery were satisfied with their decision, although satisfaction with reconstruction was mixed. CONCLUSION: Factors influencing surgical decision making may include breast-cancer-related worry, biopsy history, and subjective breast cancer risk. PMID- 7479634 TI - [An affinity chromatography sorbent containing concanavalin A groups immobilized by complex formation with cobalt]. AB - A Spheron-base affinity sorbent containing Concanavalin A (Con A) groupings has been synthesized. Con A was immobilized via the formation of a ternary complex with a stationary ligand (tetrafunctional) triethylene tetramine) and cobalt(III) ions. The inertness of cobalt(III) provided strong binding of Con A to the matrix. The sorbent was tested for the ability to absorb proteins (albumin, lysozyme, heparin, and ribonuclease) and used the isolation of glucosaminidase from the proteolytic complex of the fungus Geotrichum candidum. The purified enzyme was characterized biochemically. PMID- 7479635 TI - [Regulation of biosynthesis of intracellular enzymes in Bacillus intermedius 3S 19]. AB - The addition of microelements (Co2+, Cu2+, Zn2+, and Fe2+) to a cultivation medium increased the activity of phosphomonoesterase but not of proteinase and ribonuclease. Glucose and inorganic phosphate (Pi) were the main factors that affected the direction and intensity of the biosynthesis of extracellular enzymes. PMID- 7479636 TI - [Interrelationship between populations of cellulolytic microorganisms during digestion of cellular tissue by rumen contents]. AB - Digestion of blotting paper and straw by microorganism contained in the rumen of cattle was studied. It was established that bacterial populations (with inhibition of fungi by nystatin) and fungal populations (with inhibition of bacterial by penicillin and streptomycin) digested a greater amount of cellular tissue than the native rumen contents did. Counting the bacterial and fungal populations suggested the existence of competitive relationships between the cellulolytic microorganisms of the rumen. PMID- 7479637 TI - [The effect of autolysis on characteristics of amino acid mixtures, obtained using ethanol-assimilating yeasts]. AB - The influence of the enzyme preparation - macerate, concentration of the yeast biomass in the reaction medium, and chloroform on the autolysis of the ethanol assimilating yeast Saccharomyces cerevisiae BKM-Y-2656, BKM-Y-2465, and the baker's yeast r. L-2 was studied. Amino acid mixtures were isolated from the yeast autolysates. The use of macerate and a decrease in the biomass concentration were shown to increase the yield of the amino acid mixtures. The use of chloroform positively influenced the baker's yeast autolysis. Conditions of the joint action of these factors on the intensification of the process of the amino acid mixtures preparation based on the autolysis were determined. PMID- 7479638 TI - [Does AIDS influence the epidemiology of tuberculosis?]. PMID- 7479639 TI - [Patient Education Study Group of the German Society of Pneumology and the German Respiratory League of the German Society of Pneumology. Recommendations for structuring patient training in obstructive respiratory tract diseases]. PMID- 7479640 TI - [Adjusting background flow in measuring ventilation of newborn infants and infants using the flow-through technique]. AB - Ventilatory measurements especially in preterm infants are hampered by the additional apparatus dead space (face mask, pneumotach, shutter, ...). The apparatus dead space can be higher than the physiological dead space and affects gas exchange and blood gases significantly. For lung function testing in premature or term infants a background flow (flow-through technique) is frequently used to eliminate the apparatus dead space. This background flow must be higher than the maximum inspiratory flow (maxVin). For its optimization ventilatory measurements were performed in 62 sleeping infants aged 2 to 28 days (birth weight 760-5000 g). maxVin correlated strongly with minute ventilation VE (r = 0.90; p < 0.001). The ratio maxVin/VE decreased significantly (p < 0.05) with increasing frequency: from 4.4 +/- 0.8 (f < 40/min); 4.1 +/- 0.7 (f = 40 59/min); 3.8 +/- 0.5 (f = 60-79/min) to 3.4 +/- 0.4 (f > 80/min). With increasing frequency the flow signal becomes sinusoidal and the ratio maxVin/VE tends to pi. Therefore, during tidal breathing the background flow should be at least three but not more than the six times VE to avoid an impairment of measurements. PMID- 7479641 TI - [The importance of lung function as a predictor of 2-year mortality in mucoviscidosis]. AB - Prediction of mortality in cystic fibrosis (CF) has become an important issue in the era of lung transplantation. A study from Toronto has shown a forced expiratory volume in one second (FEV1) below 30% predicted to be the best predictor of two-year mortality. The purpose of this investigation was to assess whether these data are relevant for other CF centers with a different overall life expectancy. We have analysed pulmonary function tests (PFTs) obtained in our population of CF patients between 1979 and 1991. 19 of the 73 patients included in this analysis had died. Cox proportional hazard regression analysis was performed using the lung function parameters as single covariates. In addition results of PFTs obtained two years before death were compared to similarly lagged values of surviving patients. Inspiratory vital capacity (IVC) (p < 0.0001), the ratio of residual volume/total lung capacity (RV/TLC) (p < 0.0001), forced expiratory volume in one second (FEV1) (p < 0.0002), specific conductance (sGaw) (p < 0.025) and a weight for height below the third percentile (p < 0.023) were significant predictors of two-year mortality. Two-year mortality were 50% for a FEV1 < 30% predicted and 47% for a RV/TLC ratio > 50%. Our data confirm that a FEV1 < 30% and a RV/TLC ratio > 50% are significant predictors of mortality in CF. However, interindividual variability in survival with a FEV1 < 30% is high. The decision for lung transplantation can therefore not be based on pulmonary function alone. PMID- 7479642 TI - [The "neuron-specific enolase" tumor marker as a prognostic indicator in small cell bronchial carcinoma]. AB - To assess the predictive value of the tumour marker NSE in respect of survival time prognosis in patients with small-cell bronchial carcinoma we performed serial measurements of the NSE concentration in 67 patients in whom the small cell bronchial carcinoma had been newly diagnosed, before and during chemotherapy or radiotherapy. Pretherapeutic NSE determination proved an important predictive parameter with regard to survival time prognosis. In patients with an initial NSE concentration of over 60 ng/ml the survival time was significantly reduced (from 12.1 months to 8.4 months, p < 0.01). The pretherapeutic NSE concentrations are associated with the initial tumour stage. Serial NSE determinations reflect the course of the disease. A drop in NSE concentrations during the early phase of therapy may be important as a secondary prognosis indicator. PMID- 7479643 TI - [Cor triatriatum--a rare cause of pulmonary hemosiderosis]. AB - Pneumological examinations including open lung biopsy performed on a male patient of 30 years of age suffering from severe respiratory distress that disabled him, as well as from massive recurring attacks of hemoptysis, resulted in suspicion of idiopathic pulmonary hemosiderosis (also known as Ceelen-Gellerstedt's syndrome). Diagnosis of cor triatriatum followed by surgery was arrived at only after a pulmonary oedema had developed and after other rare cardiac diseases had been considered. This rare congenital malformation--which occasionally becomes clinically manifest only in the adult--should be suspected in differential diagnosis of respiratory distress and a sometimes also life-threatening hemoptysis. Echocardiography is the diagnostic method of choice in this regard. PMID- 7479644 TI - Time course of the effect of praziquantel on Schistosoma mansoni attachment in vitro: comparison with its effects on worm length and motility. AB - We report herein that praziquantel inhibits the capacity of adult male Schistosoma mansoni to attach to the bottom of a glass dish and produces a diminution of worm length and motility, acting under a similar concentration and time dependency. After removal of 1 microM praziquantel from the medium, the worms progressively recuperated their initial length and motility without recovering their attachment capability. The absence of calcium or presence of 1 microM verapamil did not change the praziquantel-induced effect on worm attachment, causing only a transient decrease in worm length and motility. The present data indicate that the diminution of motility induced by praziquantel results from the progressive contraction of the longitudinal musculature of the worm. In contrast, the loss of attachment should not be causally related to the contraction of the worm since these two praziquantel-induced effects have distinct patterns of response under some experimental conditions. PMID- 7479646 TI - An immunocytochemistry study comparing the occurrence of neuroactive substances in the nervous system of cercariae and metacercariae of the eye fluke Diplostomum spathaceum. AB - The nervous system of two larval stages (cercariae, metacercariae) of eye fluke Diplostomum spathaceum was investigated immunocytochemically by the application of antisera to the amino acid glutamate and to neuropeptides isolated from invertebrates (Mollusca) and from vertebrates to whole-mount preparations. In cercariae, positive immunoreactivity (IR) was observed with antisera raised against Catch-relaxing peptide (CARP), FMRFamide, alpha-caudodorsal cell peptide (alpha-CDCP), substance P, vasotocin, and vasopressin. In metacercariae, in addition to positive staining with these antisera, the ones raised against glutamate, APGWamide, caudodorsal cell hormone I (CDCH-I), and small cardiac peptide B (SCPB) also gave positive IR in the nervous system. In the two larval stages the most extensive pattern of IR was observed with anti-FMRFamide and anti CARP. In the nervous system of metacercariae the same immunoreactive neurosubstances appeared to be present as in that of cercariae. The increase in the variety of immunoreactive neurosubstances in the more complex nervous system of metacercariae is discussed in relation to parasite development and to host adaptation. PMID- 7479645 TI - Content of some microelements in chickens with ascaridiasis under combined drug treatment. AB - The content of zinc, cobalt, and selenium in the intestinal wall of Leghorn chickens infected with Ascaridia galli was determined by nondestructive neutron activation analysis. A comparison was made between the infected animals that were treated with Piperathrine and those that were given microelements or those that did not receive microelements. Age-dependent changes were found in the content of the microelements in the control groups. It is concluded that zinc, cobalt, and selenium are unevenly distributed in the intestinal tissues, with a dominating density of selenium occurring in the epithelial layer of the intestinal mucosa. PMID- 7479648 TI - DNA measurements and ploidy determination of developmental stages in the life cycles of Theileria annulata and T. parva. AB - The relative DNA levels of different developmental stages of Theileria annulata and T. parva in the cow and the tick were measured by the cytophotometric DNA technique using the fluorochrome Hoechst 33258 as a staining dye. The results revealed that sporozoites, merozoites, gamonts, and gametes were haploid, whereas multinucleated intralymphocytic schizonts were polyploid. No difference was observed between T. parva and T. annulata in these stages. For both Theileria species, the DNA measurements revealed that fusion of gametes occurred in the gut of the final host, thus providing evidence of sexual reproduction. However, differences were observed between the two parasites in the tick. Whereas T. parva zygotes underwent a two-step meiotic division, a comparable reduction division could not be unequivocally detected in T. annulata. Differences could also be detected in the further development of kinetes, indicating that Theileria species are not characterized by only one life cycle, which is specific for this genus. PMID- 7479647 TI - Circulating trans-sialidase activity and trans-sialidase-inhibiting antibodies in Trypanosoma cruzi-infected mice. AB - Parasite-derived trans-sialidase (TS) activity was demonstrated in the serum and blood of Trypanosoma cruzi-infected mice. Serum TS activity levels correlated well with parasitemia in BALB/c and Swiss mice during the initial stages of the infection. However, in later stages the TS activity levels decreased despite increasing parasitemia. This coincided with the appearance of circulating TS antibodies. On the other hand, there was always a good correlation between TS activity and parasitemia in athymic nude mice. Sera from mice with high parasitemia and low TS activity inhibited TS activity in vitro. The inhibition was also observed with purified serum IgG, and it was absorbed by staphylococcal protein A, indicating that it was caused by anti-TS IgG antibodies. These antibodies inhibited the enzymatic activity of insolubilized TS, indicating that they act by interfering with the catalytic site rather than by aggregating the enzyme. The presence of inhibitory antibodies, however, did not prevent the progression of parasitemia in BALB/c mice. PMID- 7479649 TI - Isolation, in vitro culture, ultrastructure study, and characterization by lectin agglutination tests of Phytomonas isolated from tomatoes (Lycopersicon esculentum) and cherimoyas (Anona cherimolia) in southeastern Spain. AB - Plants of Lycopersicon esculentum (grown in greenhouses) and Anona cherimolia cultivated in southeastern Spain were examined for the presence of trypanosomatid flagellates. Kinetoplastid protozoa were found in the fruits but not in the phloem or other plant tissues. Parasites were detected from the onset of fruiting. Isolates were detected from the onset of fruiting. Isolates were adapted to in vitro culturing in monophase media. The form and the structural organization was studied by scanning and transmission electron microscopy. The parasites showed an ultrastructural pattern similar to that of other species of the genus Phytomonas. In tomatoes experimentally inoculated with flagellates cultivated in vitro, we observed that the parasites did not lose their infectious capacity. Three strains of trypanosomatids of the genus Phytomonas, isolated from different species of Euphorbia (E. characias and E. hyssopifolia) and from Cocos nucifera, were compared with our isolates by lectin-agglutination tests. Our isolates were different from the two strains isolated from Euphorbia, but with this technique we could not differentiate our isolates from those of the coconut, nor could we differentiate between the isolates, their ultrastructural similarity together with their similar behavior in the lectin-agglutination test suggesting that these isolates have a common origin. PMID- 7479650 TI - Disaccharidase deficiencies in Mongolian gerbils (Meriones unguiculatus) protected against Giardia lamblia. AB - The activities of the disaccharidases lactase, maltase, sucrase and trehalase were examined in gerbils during Giardia lamblia infections. In a primary infection with trophozoites, the activities of all four enzymes were reduced from day 10 post-infection (p.i.) and remained at low levels well past the elimination phase of the infection. However, during a challenge infection, the disaccharidase decreases were short-lived, with impairments being seen only on days 2 and/or 4 post-challenge (p.c.). Sucrase activity was not affected by a challenge infection. When 0.1 mg of a soluble extract of G. lamblia trophozoites was used to challenge gerbils previously exposed to the live parasite, the pattern and duration of enzyme deficiencies were comparable with those observed after the challenge with the live parasite. In addition, decreasing the extract dose used to challenge the gerbils led to smaller disaccharidase deficiencies. G. lamblia infected gerbils were also challenged with a soluble extract of Entamoeba histolytica trophozoites, and this had no effect on the disaccharidase activities. Therefore, the presence of the intact parasite was not necessary to induce enzyme reductions in immune animals. In addition, the effects seen during the secondary infection were parasite-specific and may have involved the host's immune response to Giardia antigens. Immune gerbils were further challenged with the in vitro-released excretory/secretory products of G. lamblia. Under our experimental conditions, disaccharidase activities were found to be affected by these products in a manner that was inconsistent with the results of the live parasite challenge, and this merits further study. PMID- 7479653 TI - Immunology and morphology studies on the proliferation of in vitro cultivated Echinococcus multilocularis metacestodes. AB - The larval stage of Echinococcus multilocularis causes alveolar echinococcosis (AE) in various mammals, including humans. Traditionally metacestodes are maintained in the laboratory by serial transplantation passages into susceptible animals such as mice or gerbils. However, in animal models it has always been difficult to draw definite conclusions about the factors modulating metacestode differentiation, and investigations on gene expression and respective regulation have been hampered by the complexicity of the host-parasite interplay. This paper describes the maintenance and proliferation of E. multilocularis metacestodes as well as the formation of protoscolices in a chemically defined medium devoid of host influence. The interactive role of a heterologous human cell line (CACO2) in the in vitro development of metacestodes was also assessed. The morphology and ultrastructure of in vitro-generated metacestodes was studied using scanning (SEM) and transmission electron microscopy (TEM). Different cultivation procedures were analyzed in terms of expression of B- and T-cell epitopes and of the relevant laminated layer-antigen Em2; the exact localization of this antigen was further demonstrated by immunogold electron microscopy. PMID- 7479651 TI - Evidence of two genetic entities in Bothriocephalus funiculus (Cestoda) detected by arbitrary-primer polymerase chain reaction random amplified polymorphic DNA fingerprinting. AB - The genetic diversity of two samples of Cestoda (Bothriocephalus funiculus, Renaud and Gabrion, 1984) parasitizing two sympatric teleostean species was assessed using random amplified polymorphic DNA (RAPD). A total of 72 Bothriocephalus were analyzed individually, and electrophoretic analysis of the amplification products of 65 primers among the 68 tested revealed monomorphic patterns, reflecting the close genetic relatedness within and between the parasites of the two samples. However, 3 primers showed polymorphic patterns at 6 RAPD sites. Analysis of the distribution of these genomic fragments, assuming random mating, showed strong linkage disequilibria (only 8 genetic combinations were observed among the 32 expected). Two genetic entities displaying a high degree of host specificity were evidenced within our two samples of funiculus. This powerful molecular technique can be used as a diagnostic tool in studies concerning the biodiversity of related genetic entities and could have broad applications in parasitology. PMID- 7479652 TI - DNA measurements reveal differences in the life cycles of Babesia bigemina and B. canis, two typical members of the genus Babesia. AB - The relative DNA levels of different developmental stages of Babesia bigemina and B. canis were measured by cytophotometry using the fluorochrome Hoechst 33258 as a staining dye. The DNA measurements provided direct proof of sexual reproduction, i.e., fusion of gametes resulting in the formation of zygotes followed by a meiotic division. Both Babesia species are considered to be typical members of the genus Babesia; however, the DNA measurements revealed important differences in the life cycle of these parasites, indicating that Babesia species are not characterized by a life cycle, which is specific for this genus. PMID- 7479654 TI - In vitro effects of VD-99-11 on Angiostrongylus cantonensis and isolated frog rectus. AB - In vitro effects of VD-99-11 were examined using adult Angiostrongylus cantonensis and isolated frog rectus. In A. cantonensis, paralysis was elicited by VD-99-11 at 10(-9)-10(-6) g/ml. The paralysis caused by VD-99-11 (10(-8) g/ml) was antagonized by picrotoxin or bicuculline but not by phentolamine. A relationship between VD-99-11 and gabergic antagonists was observed in worm preparations contracted by eserine or pyrantel: VD-99-11 at higher concentrations (3x10(-6) g/ml) caused a marked contraction. In worm preparations contracted with eserine or pyrantel, the only additional contraction induced by VD-99-11 (5x10( 6) g/ml) was antagonized by strychnine. In experiments on the guanidine (2.5x10( 3) M)-induced twitch responses in isolated frog rectus, marked stimulation was caused by VD-99-11 (3-5x10(-6) g/ml). The stimulated responses induced by VD-99 11 were antagonized by tetrodotoxin, D-tubocurarine, strychnine, and hemicholinium-3, respectively. These results suggest that VD-99-11 seems superior to milbemycin D, milbemycin oxime, and ivermectin in some aspects, such as in vitro potency, though this new substance is similar to these drugs in having two different actions on the gabergic mechanism at lower concentrations and on the cholinergic mechanism at higher concentrations. PMID- 7479655 TI - 3'-Deoxy-3'-fluoroinosine as a potent antileishmanial agent. The metabolism and selective cytotoxic effect of 3'-deoxy-3'-fluoroinosine against Leishmania tropica and L. donovani in vitro and in vivo. AB - We studied the antileishmanial activity of 3'-deoxy-3'-fluoroinosine (3'-FI) against Leishmania tropica and L. donovani. In in vitro cultivation, the EC50 values (the concentration of drug necessary to inhibit the growth rate of cells to 50% of the control value) obtained for 3'-FI against the promastigotes of L. tropica and L. donovani were 2.3 x 10(-7) and 1.0 x 10(-6) M, respectively. It was less toxic toward mouse mammary-tumor FM3A cells, a model host; the EC50 value was 1.9 x 10(-4) M. Leishmania promastigote metabolized 3'-FI to 3'-deoxy 3'-fluoroadenosine 5'-triphosphate (3'-FATP) but FM3A cells did not. 3'-FI was effective against L. donovani amastigotes in J774.1 cells in an in vitro cultivation system under conditions similar to those used in the in vivo assay. 3'-FI (50 mg/kg, given i.v.) showed a cytotoxic effect against the amastigotes of L. donovani in mice. PMID- 7479656 TI - Tetraploids of the lung fluke Paragonimus westermani found in China. AB - Two groups of Paragonimus westermani (Tematoda: Platyhelminthes) exist in nature: diploids and triploids. Generally, these two groups live allopatrically, but in Kuandian, Liaoning Province, in the Republic of China, they live sympatrically. In our Chinese experiment on Paragonimus we used metacercariae of P. westermani, which we collected in Kuandian, Xigutai, and performed a cytological analysis. The results were as follows: (1) the P. westermani in Xigutai lived sympatrically as diploids and triploids; (2) all of the small metacercariae were diploids; (3) the large metacercariae were in large proportion triploids; (4) we found one tetraploid specimen in both the medium and the large metacercariae--this was the first time tetraploid lung flukes were discovered; (5) the somatic chromosomes of the tetraploids were different in numbers (4n = 44), but we could not find any difference in the karyotype of haploid sets and that of the diploids and the triploids; (6) unlike the triploids, during their meiosis the tetraploids produced a chromosome pairing, and we found a tendency of the large chromosomes to become quadrivalent; and (7) also unlike the triploids, a great number of spermatids were found in the tetraploid testes. Because of these findings, we can consider tetraploids to be autotetraploids, and these are probably produced by the fertilization of diploids and triploids. We also think that the gametes of tetraploids have a fertilization capability. PMID- 7479657 TI - Factors related to driving in persons with an implantable cardioverter defibrillator. AB - A study of 97 persons (mean age = 66 yrs, 79% male) with an ICD for an average of 2.2 years was conducted to determine whether patients resume driving (N = 72) post-ICD insertion despite instructions not to do so. Those who had resumed were queried about their driving habits, the presence of symptoms associated with arrhythmias, the occurrence of shocks in the previous year, and the importance of driving to maintenance of lifestyle. Our assumption was that patients return to driving to maintain their pre-ICD lifestyle of functional independence, and to resume social roles such as provider for the family. Seventy-four percent of subjects reported driving an average of 60 mi/week despite being instructed not to drive by their physician or other health care provider. Of those who resumed driving, > 4% had received a shock while driving. Over 86% of subjects believe driving was an important part of maintaining one's lifestyle. Reasons for driving included necessity (62%), such as to work or a physician appointment, or social (58%), such as driving to the store or church. Symptoms such as dizziness, palpitations and lightheadedness were experienced by 80% of subjects, with 43% receiving a shock from their ICD within the previous year. There were significant correlations between driving and the importance of driving to maintaining one's lifestyle (p < .05), driving for necessity (p < .01), for social reasons (p < .01) and being the primary driver in the family (p < .05). PMID- 7479658 TI - Angina, functional impairment and physical inactivity among Mexican-American women with depressive symptoms. AB - There has been a long-standing interest in the relationship between cardiovascular disease (CVD) and psychiatric morbidity, specifically depressive symptoms. The aims of this study were to determine the following: 1. The relationship between the presence of angina and depressive symptoms; 2. The influences of sociodemographic covariates on this relationship; and 3. The effects of functional impairment or inactivity level in mediating the relationship between angina and depressive symptoms. The data utilized for this study was from the Hispanic Health and Nutrition Examination Survey (HHANES) on 1,558 Mexican American women. The survey was cross sectional, conducted in 1982 84. The dependent variable was depression as measured by the Center for Epidemiologic Depression Scale (CES-D). The independent variable was the presence of angina as measured by the Rose Questionnaire. Covariates included functional impairment, physical activity, age, education, acculturation, poverty level, marital status, and work status. The findings suggest that functional impairment has a stronger association with depressive symptoms than angina or physical inactivity. Significant sociodemographic variables include single marital status and poverty. Comorbidity has been a neglected area of research for Mexican American women, but this study suggests that functional impairment is a more important factor associated with depressive symptomatology than angina. PMID- 7479660 TI - Pain, pain relief and accuracy of their recall after cardiac surgery. AB - This study investigated how much pain and pain relief cardiac surgery patients experience in Intensive Care Units (ICUs), and the accuracy of their recall later, during hospital recovery. Thirty-nine patients completed the first interview in the ICU, and 31 of them answered recall questions later. The worst pain patients experienced in the ICU was moderately high, and patients reported 65% pain relief from analgesics administered. In general, patients' later recalled ICU pain was less severe and recalled pain relief from pain medications was greater than what they had reported when in the ICU. A substantial number of patients were unable to rate the effects of analgesics because they did not remember being mediated. One gender difference was noted: women recollected having a higher "worst pain" in ICU than men. Although recall of ICU pain is less than accurate, ICU cardiac surgical patients have episodes of substantial pain. Despite advances in pain management, patients report that pain relief is frequently incomplete and that they do not remember receiving analgesics. A more proactive approach to pain management by health care professionals, which includes informing patients when they are receiving analgesics, may help to improve pain relief in cardiac surgical patients. PMID- 7479659 TI - Behavioral and physiological response during feeding in infants with congenital heart disease: a naturalistic study. AB - Twenty mother-infant dyads, ten with congenital heart disease (CHD) and ten controls, were studied during a naturally occurring feeding episode. Heart and respiratory rate data from a Hewlett Packard Infant Monitor were recorded on a computer while the feeding was videotaped for later scoring. No significant relationship was found between potent disengagement cues and changes in either heart or respiratory rate during feeding. However, the presentation of subtle disengagement cues were observed to be significantly more frequent among infants with CHD. A wide range of individual differences were noted in presentation of disengagement cues and heart and respiratory rates in both groups. No discernable pattern of heart rate or respiratory rate increase or decrease related to subtle or potent disengagement cures was found in either the infants with CHD or the healthy controls. PMID- 7479661 TI - Pharmacologic management of cardiac arrest: an update perspective. PMID- 7479662 TI - Physician education and management of hypercholesterolemia and compliance issues. PMID- 7479663 TI - Shivering associated with cardiac surgery: predictive variables and impact on nursing intervention. PMID- 7479664 TI - Regular atrial fibrillation spells trouble. PMID- 7479665 TI - Atrial fibrillation and flutter after coronary artery bypass surgery: impact on recovery. PMID- 7479666 TI - Lazaroids protect isolated rat pancreatic acinar cells against damage induced by free radicals. AB - Lazaroids, 21-aminosteroids without gluco- and mineralocorticoid activity, protect against oxidative injury in nervous system cells and may therefore also have a potential for treatment of pancreatitis, where oxidative stress contributes to cell injury. The present study evaluates the protective potential of the lazaroids U-78518F, U-74500A, and U-74389F against damage to isolated pancreatic acinar cells exposed to two models of oxidative stress: (a) a XOD/HX model, consisting of xanthine oxidase, hypoxanthine, and chelated FeCl3; and (b) an ADP/Fe model, consisting of FeSO4 and the reducing agent ADP. Both models caused time-dependent cell injury as assessed by uptake of trypan blue and release of lactate dehydrogenase. Short-term peak production of free radicals in the XOD/HX model--as monitored by the deoxyribose assay--was more injurious to cells than continuous radical generation at lower levels in the ADP/Fe model. In general, lazaroids at 1-10 microM reduced oxidative damage and deoxyribose oxidation in both models. The degree of reduction of cell damage and deoxyribose oxidation depended on the type and concentration of the lazaroid and the model used. Lazaroid concentrations < 0.1 microM were ineffective, and concentrations > 50 microM even accelerated cell injury, although lazaroids still served as scavengers at high concentrations. At least part of the noxious effects of high lazaroid concentrations is due to nonspecific membrane damage because these concentrations caused cell injury also in the absence of oxidative stress. The limited range of protective concentrations has to be observed in further in vivo studies. Interestingly, acinar cells in the absence of lazaroids also reduced radical-induced deoxyribose degradation.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7479667 TI - Detection of superoxide free radicals in rats with acute pancreatitis. AB - To study the importance of oxygen-derived free radicals in acute pancreatitis, an experimental study of in vivo detection of superoxide free radicals (O2-) was performed using rats. Using a new chemiluminescence probe (2-methyl-6-[p methoxyphenyl]-3,7-dihydro-imidazol[1,2-1]pyrazin- 3-one; MCLA; a Cypridina luciferin analogue) and a highly sensitive photon counting system, O2- from the pancreatic surface of rats with experimental acute pancreatitis induced by 180 micrograms cerulein/kg was detected. The time course of MCLA-dependent luminescence suggested that O2- production began 2-3 h after cerulein injection and then decreased gradually. Superoxide free radical production in the pancreas of rats with cerulein-induced acute pancreatitis was confirmed using MCLA dependent chemiluminescence. This new method allows direct observation of the behavior of oxygen-derived free radicals. PMID- 7479668 TI - The effects of green tea catechins (Polyphenon) on DL-ethionine-induced acute pancreatitis. AB - The effects of green tea catechins (Polyphenon) on DL-ethionine-induced acute pancreatitis in rats were examined. The acute pancreatitis induced in this study was characterized by moderate inter- and intrastitial edema and patchy acinar cell necrosis. In rats induced with acute pancreatitis by an intraperitoneal injection of DL-ethionine, the wet weight of the pancreas (0.47 +/- 0.059 g/100 g body weight; p < 0.05), the serum amylase (10,432 +/- 996 IU/L; p < 0.001), and the tissue concentration of lipid peroxides (19.5 +/- 1.78 nmol/mg tissue DNA; p < 0.001) were significantly increased compared with values obtained in control rats (0.39 +/- 0.037 g/100 g body weight, 5,639 +/- 1,568 IU/L, and 10.7 +/- 1.04 nmol/mg tissue DNA, respectively) injected with isotonic saline. In contrast, in rats injected with DL-ethionine and supplied with a green tea catechin solution as a beverage instead of water during the experimental period, the tissue of pancreas was almost-correct, and the wet weight of the pancreas (0.39 +/- 0.054 g/100 g body weight; p < 0.05), the serum amylase (5,716 +/- 708 IU/L; p < 0.001), and the concentration of lipid peroxides in tissue (11.5 +/- 2.15 nmol/mg tissue DNA; p < 0.001) were significantly decreased compared with values obtained in rats injected with DL-ethionine and supplied with water as a beverage. These data suggest that green tea catechins may have a protective effect on the pathogenesis of experimental acute pancreatitis. PMID- 7479669 TI - Therapeutic regimens in acute experimental pancreatitis in rats: effects of a protease inhibitor, a beta-agonist, and antibiotics. AB - The osmolality of contrast injected retrograde into the rat pancreatic duct did not affect the severity of the pancreatitis (Urografin, 1,300 mOsm/kg, and Hexabrix, 580 mOsm/kg). The severity of the pancreatitis induced in rats was assessed by survival rate, histologic grading, wet lung ratio, and serum levels of amylase, lipase, and trypsin-like activity. Rats with pancreatitis induced by retrograde injected Urografin, lipopolysaccharide, taurocholic acid plus enterokinase were treated with either intravenous (i.v.) FUT-175 (Nafamstat Mesilate), FUT-175 administered by retrograde pancreatic injection, i.v. terbutaline, i.v. piperacillin sodium, piperacillin sodium by retrograde pancreatic duct injection, or a combination of FUT-175 plus terbutaline and piperacillin. Survival among the rats was increased and the incidence of pancreatic infection reduced in rats treated with i.v. piperacillin or with a combination of FUT-175 plus i.v. terbutaline, plus i.v. piperacillin compared to controls. PMID- 7479670 TI - Energy metabolism in mouse pancreas in response to different dosages of a CCK analogue. AB - Stimulation of the exocrine pancreas with cholecystokinin analogues leads to a variety of intraacinar processes, many coupled to energy consumption. It was hypothesized that extensive ATP depletion could play a role in the pathophysiology of acute pancreatitis, especially in the hyperstimulation (cerulein) model. Mice received seven intraperitoneal injections of cerulein at hourly intervals, at doses ranging from physiological (0.1 micrograms/kg) to pharmacological (50 micrograms/kg). A single dose of cerulein induced a 28-33% decrease in ATP, whereas a complete course of injections led to a nadir as low as 45% of the control value. The overall pattern of ATP tissue content during the observed time course was surprisingly similar in all four groups and statistically not different at any time point. Until 12 h, ATP levels in all groups remained below the control value. In contrast, serum amylase and light microscopy reflected a degree of pancreatitis in a close dose-response pattern to the administered cerulein dose. These findings suggest that ATP depletion- although probably facilitating acinar damage--does not seem to play a causal or primary role in the pathophysiology of acute pancreatitis. PMID- 7479671 TI - Changes in gene expression of pancreatitis-associated protein and pancreatic secretory trypsin inhibitors in experimental pancreatitis produced by pancreatic duct occlusion in rats: comparison with gene expression of cholecystokinin and secretin. AB - Pancreatic duct occlusion is known to produce a sustained increase in the plasma cholecystokinin (CCK) concentration and to affect the tissue content of CCK in the rat. The tissue content of CCK is correlated with regenerative changes in the pancreas after pancreatic duct occlusion. In the present study, we examined the changes in mRNA levels of pancreatic secretory trypsin inhibitors (PSTIs), pancreatitis-associated protein (PAP), and amylase in the pancreas in comparison with changes in CCK and secretin mRNA levels in the intestine and the histological changes produced by pancreatic duct ligation. Rats with an internal bile fistula and with obstruction of pancreatic flow were prepared and were sacrificed 1, 3, 7, 10, 14, and 28 days later. Then mRNA levels of CCK, secretin, PSTIs, PAP, and amylase were determined by slot-blot analysis. The CCK mRNA level gradually increased to a peak on day 10, was slightly lower on day 14, and returned to the control level on day 28. The level of secretin mRNA did not change. The mRNA levels of PSTIs increased significantly on day 3 after occlusion. PAP mRNA was detectable on days 1 and 3, being maximal on day 1. The mRNA level of amylase was markedly decreased on days 1 and 3, then remained lower than the control level. Histological examination showed acute inflammatory changes in the pancreas on days 1 and 3 and regenerative changes from day 7. These results suggest that a change in gene expression of PAP reflects acute inflammatory changes in the pancreas most sensitively. PMID- 7479672 TI - Correlation of nucleolar organizer regions with secretory and regenerative process in experimental cerulein-induced pancreatitis in the rat. AB - Silver staining of nucleolar organizer regions (AgNORs) demonstrates loops of DNA that transcribe ribosomal RNA. Their number and size have been attributed to rDNA transcription activity involved in protein synthesis and thus associated with proliferation. The exact relationship among proliferation, protein synthesis, and expression of AgNORs is, however, not yet well established. We therefore investigated AgNORs in an experimental model of cerulein-induced rat pancreatitis. During secretory stimulation with maximal doses of cerulein (0.25 micrograms/kg/h) for 12 h, AgNOR number and size per nucleus as well as 3H thymidine label index did not change, although there was a marked increase in pancreatic volume flow, up to 150%, and of protein synthesis rate, up to 180% of the control levels. In contrast, after infusion of supramaximal doses of cerulein (5.0 micrograms/kg/h), AgNOR and 3H-thymidine label values rose significantly, with a distinct peak at day 3 after induction of pancreatitis. Most interestingly, AgNOR number and size were elevated 12 h before DNA replication started as determined by 3H-thymidine incorporation. At the same time intracellular protein synthesis proved to be decreased approximately 30-50% compared to controls. Our data confirm that AgNOR is a marker of proliferation that reflects regulatory events in the cell cycle earlier than 3H-thymidine incorporation. Here we demonstrate for the first time that this phenomenon is independent of the total intracellular protein synthesis rate. PMID- 7479673 TI - Effect of a platelet-activating factor antagonist on pancreas perfusion after 24 h of ischemia. AB - Platelet-activating factor (PAF) is a strong mediator of inflammation that is present in many mammalian tissues and cell types. In the pancreas, PAF can be synthesized in acinar cells after stimulation with secretagogues. The present study uses a perfused porcine pancreas model to investigate the role of PAF in pancreatic ischemia and the effect of the PAF antagonist bepafant on pancreas preservation. Pancreata were preserved with or without bepafant, stored for 24 h at 4 degrees C, and then reperfused at 37 degrees C in a perfusion chamber. Reperfusions were significantly improved by the addition of bepafant. This was indicated by a significantly increased arteriovenous volume flow (16.54 +/- 1.88 ml/min versus controls 8.54 +/- 1.31 ml/min; p = 0.0068; bepafant, n = 7; controls, n = 12) and a reduced vascular resistance (p = 0.0068; bepafant, 1.95 +/- 0.22 mm Hg * min/ml versus controls 4.08 +/- 0.56 mm Hg * min/ml). Radioimmunological quantification of PAF in pancreatic tissue revealed that PAF levels remain unchanged during storage in a cold protective solution at 4 degrees C but increase significantly during surgical pancreas preparation under general anesthesia (from 142.1 +/- 21.2 to 368.8 +/- 52.5 pg/g; n = 15; p = 0.0007). The present study shows that bepafant improves pancreas preservation after cold ischemia. The beneficial effect might be explained by antagonizing inflammatory and vasoconstrictory responses to PAF synthesized during surgical pancreas preparation. PMID- 7479674 TI - Effect of chronic administration of hydrocortisone on the induction and evolution of acute pancreatitis induced by cerulein. AB - The effects of chronic administration of hydrocortisone (10 mg/kg/day) on the development and evolution of acute pancreatitis induced by supramaximal stimulation with cerulein were examined in the rat. In these circumstances the potentially therapeutic effect of L-364,718, a CCK-receptor antagonist, was assayed. Administration of hydrocortisone over 7 days did not increase the severity of edematous acute pancreatitis induced by cerulein, since the reduction in pancreatic secretion, the hyperamylasemia and the increase in the levels of hematocrit and fluid in the pancreatic tissue were similar in rats with acute pancreatitis treated and untreated with hydrocortisone previously. When hydrocortisone was administered chronically, before administration of supramaximal doses of cerulein, a spontaneous regression of acute pancreatitis occurred. However, when hydrocortisone administration was continued after inducing pancreatitis, pancreatic recovery was prevented, observing a significantly depressed acinar secretion and elevated values of hematocrit and tissue fluid (edema). L-364,718 administration proved to be detrimental in the evolution of edematous acute pancreatitis when the rats had been treated chronically with hydrocortisone because the blockade exerted on secretion prevented the draining of enzymes stored in excess by hydrocortisone administration. PMID- 7479675 TI - Gastric acid-independent enhancement of exocrine pancreatic secretion by dietary protein in chronic bile-pancreatic juice diverted rats. AB - Diversion of bile-pancreatic juice (BPJ) causes exocrine pancreatic hypersecretion in a fasting state, and we demonstrated that a feeding of dietary protein stimulates further secretion of pancreatic enzymes in chronic BPJ diverted rats. We determined that gastric acid secretion is associated with the stimulatory effect of dietary protein feeding on pancreatic secretion in the diverted rats. BPJ was diverted with a cannula of the common bile-pancreatic duct and returned to the upper ileum for 7 days. The BPJ-diverted rats were administered omeprazole (12 mg/kg body weight) orally and were fed a 25% casein, fat-free diet spontaneously under unrestrained conditions. The volume of BPJ was not changed after administration of omeprazole and feeding the diet. Total protein, amylase, and chymotrypsin secretions were not changed 1 h after omeprazole administration but increased gradually after feeding the diet. The secretion of these enzymes reached peak values 120-150 min after feeding, and these values were two- to threefold higher than those before feeding. We conclude that gastric acid secretion is not responsible for the stimulation of pancreatic secretion that occurs after feeding a dietary protein to chronic BPJ-diverted rats. PMID- 7479676 TI - The ratio between anionic and cationic trypsin in rat pancreas varies with CCK stimulation. AB - The effects of long-term hyperstimulation with cholecystokinin (CCK) on the pancreatic contents of anionic and cationic trypsin(ogen) and amylase were studied in the rat. Endogenous hyperCCKemia was evoked in rats by pancreaticobiliary diversion (PBD), and exogenous hyperCCKemia by continuous subcutaneous CCK infusion. In addition, the effect of continuous subcutaneous infusion of the CCK A receptor antagonist devazepide was studied. After 4 weeks blood samples were obtained for the determination of plasma CCK concentrations and the animals were sacrificed. The pancreatic glands were harvested, weighted, and extracted. The extracts were analyzed for anionic and cationic trypsin and amylase. Enzyme contents showed a large interindividual variation. The most consistent change in enzyme pattern was an increase in the ratio between anionic and cationic trypsin in animals with hyperCCKemia (PBD operated or CCK infused). Furthermore, this ratio decreased significantly in animals treated with devazepide. In conclusion, stimulation of the CCK receptor changed the ratio between anionic and cationic trypsin in the pancreatic gland, while it was reversed during blockade of the receptor. PMID- 7479677 TI - Pancreatitis and the risk of pancreatic cancer. AB - To offer quantitative evidence on the association between pancreatitis and pancreatic cancer, we analyzed data from a hospital-based case-control study conducted in northern Italy between 1983 and 1992, including a total of 362 incident cases of histologically confirmed pancreatic cancer and 1,408 controls admitted to hospital for acute, nonneoplastic, nondigestive tract disorders. Information was obtained using a structured questionnaire on sociodemographic characteristics and lifestyle habits (including tobacco and alcohol consumption) and a problem-oriented medical history, which included history of pancreatitis and age at its first diagnosis. Pancreatitis was reported by 24 (6.6%) cases and 18 (1.3%) controls, yielding an age- and sex-adjusted relative risk (RR) of 5.7 (95% confidence interval, 2.9-11.4). The risk of pancreatic cancer was appreciably higher 5 or more years after diagnosis of pancreatitis (RR = 6.9) than in the first 4 years (RR = 2.1), and in subjects below age 60 (RR = 8.3) than in elderly ones (RR = 2.6), but similar in males and females. The time-risk relationship is strongly indicative of a real relationship between pancreatitis and pancreatic cancer. After allowing for tobacco and alcohol, besides area of residence and education, the association between pancreatitis and pancreatic cancer appeared to be partly explained by such covariates (RR = 3.9); however, this may represent some degree of overadjustment if, for instance, alcohol is causally linked to pancreatitis, which, in turn, is causally related to pancreatic cancer. In terms of population attributable risk, pancreatitis would explain approximately 5% of pancreatic cancer cases. PMID- 7479678 TI - Effect of endogenous insulin on pancreatic exocrine secretion in perfused dog pancreas. AB - Canine pancreas was perfused with an intraarterial infusion of Krebs-Ringer bicarbonate solution containing 5% dog red blood cells, 0.1% bovine serum albumin, and 3% dextran at 15 ml/min, while portal effluent was continuously collected. Pancreatic juice was obtained in 15-min samples via main pancreatic duct cannulation. After a 1-h basal period, secretin and cholecystokinin-8 (CCK), at doses of 2.5 ng.min-1 each, were simultaneously infused for 10 min, with background infusion of a normal rabbit serum (NRS) or an antiinsulin serum (Anti I) in 5 ml each via a sidearm of the intraarterial catheter. The infusion of secretin and CCK resulted in a significant increase in pancreatic bicarbonate and protein secretion during the infusion of NRS, whereas the pancreatic secretory response of bicarbonate and protein was profoundly suppressed by the infusion of Anti-I in six pancreata so studied. This suppression by Anti-I coincided with significant increases in somatostatin and pancreatic polypeptide levels in portal venous effluent. In three additional pancreata, simultaneous infusions of Anti-I with antisomatostatin (5 ml) and antipancreatic polypeptide (5 ml) serum failed to inhibit the pancreatic exocrine secretion. These results indicate that secretin- and CCK-stimulated pancreatic secretion of bicarbonate and protein depends heavily on local action of insulin. The suppression by Anti-I of pancreatic secretion is mediated, in part, by local releases of somatostatin and pancreatic polypeptide. Thus, the insuloacinar axis plays an important regulatory role in pancreatic exocrine secretion in the dog, and it involves at least three islet hormones including insulin, somatostatin, and pancreatic polypeptide. PMID- 7479679 TI - The effects of glucagon-like peptide-I (GLP-I) on hormone secretion from isolated human pancreatic islets. AB - Glucagon-like peptide-I (GLP-I) is a potent incretin hormone that is now considered as a new therapeutic tool in the treatment of diabetes mellitus. In this study we characterized the effects of GLP-I on peptide hormone release from isolated human pancreatic islets. GLP-I stimulated insulin release in the presence of 10 mM glucose (2.8 mM glucose, 100%; 10 mM glucose, 166%; 10 mM glucose + 10 nM GLP-I, 222%) but had only a weak insulinotropic effect (128%) at 2.8 mM glucose. Glucagon release was inhibited by 10 mM glucose (2.8 mM glucose, 100%; 10 mM glucose, 72%) and by 10 nM GLP-I at 2.8 mM glucose (67%). Somatostatin secretion was increased by 10 mM glucose (2.8 mM glucose, 100%; 10 mM glucose, 166%). GLP-I stimulated somatostatin release in the presence of 2.8 mM glucose (172%). Pancreatic polypeptide (PP) secretion was enhanced by 10 mM glucose (2.8 mM glucose, 100%; 10 mM glucose, 236%). GLP-I induced PP release only in the presence of 2.8 mM glucose (184%). PMID- 7479680 TI - Effects of arginine on the secretion of insulin and islet amyloid polypeptide in humans. AB - Islet amyloid polypeptide (IAPP) is synthesized in islet B cells and stored in the secretory granules. We examined whether IAPP and insulin are released in parallel in humans. Arginine hydrochloride (5 g) was injected intravenously at three glucose levels in 11 healthy 58-year-old female subjects. In the fasting state (plasma glucose, 4.9 +/- 0.3 mM), serum insulin levels were 62 +/- 8 pM and plasma IAPP levels were 5.9 +/- 0.8 pM (r = 0.74, p < 0.01; insulin/IAPP ratio, 13.4 +/- 3.6). The insulin response to arginine was 426 +/- 84 pM (p < 0.001), whereas the IAPP response was 4.9 +/- 1.8 pM (p < 0.01) (r = 0.93, p < 0.001). At 16.3 +/- 0.5 mM glucose, the insulin response was increased to 1,516 +/- 325 pM (p < 0.001), whereas the IAPP response was increased to 10.4 +/- 2.8 pM (p < 0.01) (r = 0.91, p < 0.001). No further increases were seen at 35.0 +/- 2.0 mM glucose. The ratios of insulin response/IAPP response, which represent the relative secretion of the two peptides, were 169 +/- 31, 158 +/- 17, and 162 +/- 17, at the three glucose levels. Thus, the ratios of the insulin/IAPP responses to arginine were the same regardless of the glucose level, and the insulin and IAPP responses to arginine were highly correlated with each other at all glucose levels. We conclude that the two peptides are cosecreted in strict parallelism after arginine stimulation in humans over a wide range of glucose levels. PMID- 7479682 TI - The 2 A crystal structure of subtilisin E with PMSF inhibitor. AB - Using enzyme prepared by the DNA recombination technique, subtilisin E from Bacillus subtilis was crystallized in space group P2(1)2(1)2(1) with two molecules in an asymmetric unit. The crystal structure of PMSF-inhibited subtilisin E was solved by molecular replacement followed by refinement with the X-PLOR program. This resulted in the 2.0 A structure of subtilisin E with an R factor of 0.191 for 8-2 A data and r.m.s. deviations from ideal values of 0.021 A and 2.294 for bond lengths and bond angles respectively. The PMSF group covalently bound to Ser221 appeared very clearly in the electron density map. Except for the active site disturbed by PMSF binding, the structural features of subtilisin E are almost the same as in other subtilisins. The calcium-binding sites are different in detail in the two independent molecules of subtilisin E. Based on the structure, the remarkably enhanced heat stability of mutant N118S of subtilisin E is discussed. It is very likely that there is an additional water molecule in the mutant structure, which is hydrogen bonded to side chains of Ser118 and its neighbouring residues Lys27 and Asp120. PMID- 7479683 TI - The application of hydrogen bonding analysis in X-ray crystallography to help orientate asparagine, glutamine and histidine side chains. AB - Protein X-ray crystallography produces an electron density map that rarely detects individual hydrogen atoms or distinguishes between carbon, nitrogen and oxygen atoms in the electron density. This makes it difficult to orientate the side chains of Asn, Gln and His, which appear symmetrical in the electron density; their orientation is usually judged on the basis of hydrogen bonding. Based on the observation that almost all buried donors and acceptors are satisfied, we have developed a simple algorithm to compare the alternative conformations of these residues and, where possible, identify the most favourable. In a cross-section of protein structures we found a few side chains (15.0% of Asn and Gln and 9.9% of His) which would be more favourable in the alternative orientation. We have also found that this proportion rises slightly with worsening resolution. PMID- 7479681 TI - Defective glucose-stimulated insulin release from perifused islets of C57BL/6J mice. AB - Previous work has shown that the C57BL/6J (BL/6) mouse strain develops type 2 diabetes after being fed a high-fat, high-simple carbohydrate (HFHSC) diet. In contrast, the AJ mouse strain does not. The aim of the present study was to determine if differences in the insulin secretory characteristics of isolated perifused islets of these animals could help explain why the BL/6 mouse develops diet-induced diabetes. Insulin secretion was assessed as mean integrated area under the curve during 20 min of stimulation with 27.7 mM glucose or 5 mM lauric acid. We found that both glucose- and laurate-stimulated insulin secretions were significantly less in euglycemic BL/6 mice than in the euglycemic AJ mice. The defect in insulin response to glucose, but not laurate, in islets from the BL/6 mouse was exacerbated when the animals were fed the HFHSC diet. These data suggest that the BL/6 mouse has a defective insulin response to glucose, which is exacerbated by a diabetogenic diet. PMID- 7479684 TI - Neural network system for the evaluation of side-chain packing in protein structures. AB - An artificial neural network system is used for pattern recognition in protein side-chain-side-chain contact maps. A back-propagation network was trained on a set of patterns which are popular in side-chain contact maps of protein structures. Several neural network architectures and different training parameters were tested to decide on the best combination for the neural network. The resulting network can distinguish between original (from protein structures) and randomized patterns with an accuracy of 84.5% and a Matthews' coefficient of 0.72 for the testing set. Applications of this system for protein structure evaluation and refinement are also proposed. Examples include structures obtained after the application of molecular dynamics to crystal structures, structures obtained from X-ray crystallography at various stages of refinement, structures obtained from a de novo folding algorithm and deliberately misfolded structures. PMID- 7479686 TI - A simple algorithm for the calculation of multiple site titration curves. AB - A simple algorithm for the calculation of multiple site titration curves is proposed. It is based on a hybridization of two computational techniques: (i) a modified Tanford-Roxby iterative procedure [Tanford and Roxby (1972) Biochemistry, 11, 2193-2198] and (ii) the Boltzmann statistics. The sites characterized by strong electrostatic coupling were selected for statistical mechanical treatment, whereas all other sites were treated by means of the modified Tanford-Roxby procedure. The selection of the two sets was made on the basis of a criterion related to the interaction energy between the titratable sites in the protein molecule. The algorithm was tested for bovine pancreatic trypsin inhibitor and the pK values calculated were discussed in the light of experimental data and theoretical results obtained by other authors. The algorithm can easily be coded and incorporated into any program package for the calculation of electrostatic interactions in proteins. PMID- 7479685 TI - Context dependence of phenotype prediction and diversity in combinatorial mutagenesis. AB - Two different combinatorial mutagenesis experiments on the light-harvesting II (LH2) protein of Rhodobacter capsulatus indicate that heuristic rules relating sequence directly to phenotype are dependent on which sets or groups of residues are mutated simultaneously. Previously reported combinatorial mutagenesis of this chromogenic protein (based on both phylogenetic and structural models) showed that substituting amino acids with large molar volumes at Gly beta 31 caused the mutated protein to have a spectrum characteristic of light-harvesting I (LH1). The six residues that underwent combinatorial mutagenesis were modeled to lie on one side of a transmembrane alpha-helix that binds bacteriochlorophyll. In a second experiment described here, we have not used structural models or phylogeny in choosing mutagenesis sites. Instead, a set of six contiguous residues was selected for combinatorial mutagenesis. In this latter experiment, the residue substituted at Gly beta 31 was not a determining factor in whether LH2 or LH1 spectra were obtained; therefore, we conclude that the heuristic rules for phenotype prediction are context dependent. While phenotype prediction is context dependent, the ability to identify elements of primary structure causing phenotype diversity appears not to be. This strengthens the argument for performing combinatorial mutagenesis with an arbitrary grouping of residues if structural models are unavailable. PMID- 7479687 TI - Second-generation octarellins: two new de novo (beta/alpha)8 polypeptides designed for investigating the influence of beta-residue packing on the alpha/beta-barrel structure stability. AB - The sequence of octarellin I, the first de novo (beta/alpha)8 polypeptide, was revised according to several criteria, among others the symmetry of the sequence, beta-residue volume and hydrophobicity, and charge distribution. These considerations and the overall conclusions drawn from the first design led to two new sequences, corresponding to octarellins II and III. Octarellin II retains perfect 8-fold symmetry. Octarellin III has the same sequence as octarellin II, except for the beta-strands which exhibit a 4-fold symmetry. The two proteins were produced in Escherichia coli. Infrared and CD spectral analyses of octarellins II and III reveal a high secondary structure content. Non-denaturing gel electrophoresis, molecular sieve chromatography and analytical ultracentrifugation suggest that both of these second-generation artificial polypeptides exist as a mixture of a monomer and a dimer form. Octarellins II and III are at least 10 times more soluble than octarellin I. Urea-induced unfolding followed by fluorescence emission suggests that the tryptophan residues, designed to be buried in the (beta/alpha)8, are indeed packed in the hydrophobic core of both proteins. However, octarellin III displays a higher stability towards urea denaturation, indicating that introducing 4-fold symmetry into the beta-barrel might be important for stability of the overall folding. PMID- 7479688 TI - Engineering ribonuclease A: production, purification and characterization of wild type enzyme and mutants at Gln11. AB - Bovine pancreatic ribonuclease A (RNase A) has been the object of much landmark work in biological chemistry. Yet the application of the techniques of protein engineering to RNase A has been limited by problems inherent in the isolation and heterologous expression of its gene. A cDNA library was prepared from cow pancreas, and from this library the cDNA that codes for RNase A was isolated. This cDNA was inserted into expression plasmids that then directed the production of RNase A in Saccharomyces cerevisiae (fused to a modified alpha-factor leader sequence) or Escherichia coli (fused to the pelB signal sequence). RNase A secreted into the medium by S.cerevisiae was an active but highly glycosylated enzyme that was recoverable at 1 mg/l of culture. RNase A produced by E.coli was in an insoluble fraction of the cell lysate. Oxidation of the reduced and denatured protein produced active enzyme which was isolated at 50 mg/l of culture. The bacterial expression system is ideal for the large-scale production of mutants of RNase A. This system was used to substitute alanine, asparagine or histidine for Gln11, a conserved residue that donates a hydrogen bond to the reactive phosphoryl group of bound substrate. Analysis of the binding and turnover of natural and synthetic substrates by the wild-type and mutant enzymes shows that the primary role of Gln11 is to prevent the non-productive binding of substrate. PMID- 7479689 TI - Conversion of human 15-lipoxygenase to an efficient 12-lipoxygenase: the side chain geometry of amino acids 417 and 418 determine positional specificity. AB - Positional specificity determinants of human 15-lipoxygenase were examined by site-directed mutagenesis and by kinetic analysis of the wild-type and variant enzymes. By comparing conserved differences among sequences of 12- and 15 lipoxygenases, a small region responsible for functional differences between 12- and 15-lipoxygenases has been identified. Furthermore, the replacement of only two amino acids in 15-lipoxygenase (at 417 and 418 in the primary sequence) by those found in certain 12-lipoxygenases results in an enzyme that has activity similar to 12-lipoxygenase. An examination of the activity of nine variants of lipoxygenase demonstrated that the amino acid side-chain bulk and geometry of residues 417 and 418 are the key components of the positional specificity determinant of 15-lipoxygenase. Overexpression of a variant (containing valines at positions 417 and 418) that performs predominantly 12-lipoxygenation was achieved in a baculo-virus-insect cell culture system. This variant was purified to > 90% homogeneity and its kinetics were compared with the wild-type 15 lipoxygenase. The variant enzyme has no change in its apparent KM for arachidonic acid and a minor (3-fold) change in its Vmax. For linoleic acid, the variant has no change in its KM and a 10-fold reduction in its Vmax, as expected for an enzyme performing predominantly 12-lipoxygenation. The results are consistent with a model in which two amino acids of 15-lipoxygenase (isoleucine 417 and methionine 418) constitute a structural element which contributes to the regiospecificity of the enzyme. Replacement of these amino acids with those found in certain 12-lipoxygenases results in an enzyme which can bind arachidonic acid in a catalytic register that prefers 12-lipoxygenation. PMID- 7479690 TI - Mutational analysis of DNase I-DNA interactions: design, expression and characterization of a DNase I loop insertion mutant with altered sequence selectivity. AB - A mutant of bovine pancreatic DNase I containing two additional residues in a loop next to C173 has been expressed in Escherichia coli, purified and characterized biochemically. Modelling studies suggest that the inserted arginine and glutamate side chains of the modified loop sequence C173-R-E-G-T-V176 could contact the bases 3' to the cleaved bond in the major groove of a bound DNA, and that up to 10 bp could interact with the enzyme and potentially influence its cutting rate. The loop insertion mutant has an 800-fold lower specific activity than wild-type and shows overall cleavage characteristics similar to bovine pancreatic DNase I. Compared with the wild-type enzyme, the mutant shows a strongly enhanced preference for cutting the inverted repeat: (formula: see text) or close variants thereof. Unexpectedly for a minor groove binding protein, the preferred cutting sites in opposite strands are staggered by 1 bp in the 5' direction, causing the cleavage of a TA and a TT step, respectively. This finding demonstrates that the sequence context is relatively more important for the cutting frequency than the nature of the dinucleotide step of the cleaved bond, and clearly shows that base recognition is involved in determining the sequence selectivity of the mutant. The importance of the sequence 5' to the cleaved bond for the cutting rate suggests that the additional major groove contacts may require a distortion of the DNA associated with a higher energy barrier, resulting in an increased selectivity for flexible DNA sequences and a lower overall activity of the mutant enzyme. PMID- 7479691 TI - The position of the structurally autonomous kringle 2 domain influences the functional features of tissue-type plasminogen activator. AB - Tissue-type plasminogen activator (t-PA) is composed of structurally autonomous domains. From the N-terminus of t-PA, a finger-like domain (F), an epidermal growth factor-like domain (G), two kringle domains (K1 and K2) and a serine protease domain (P) can be discerned. The K2 domain of t-PA is known to be involved in lysine binding, fibrin binding and fibrin-dependent plasminogen activation. To study the functional autonomy of the K2 domain in t-PA we constructed, with the aid of a cassette t-PA gene [Rehberg et al. (1989) Protein Engng, 2, 371-377], mutant t-PA genes coding for four molecules (FGK1K2P, FGK2K1P, GK1K2P and GK2K1P) in which the K2 domain was placed in two different positions in t-PA. The DNAs of wild-type t-PA and the t-PA variants were expressed in Chinese hamster ovary cells and the recombinant proteins were purified by affinity chromatography. All molecules were expressed in their single chain form and could be converted to their two-chain form. With these molecules, lysine binding, fibrin binding and fibrin-dependent plasminogen activation were studied. All variants showed affinity for lysyl-Sepharose and aminohexyl Sepharose. Reversal of the K domains (FGK2K1P versus FGK2K1P and GK1K2P versus GK2K1P) resulted in a 23-47% weaker interaction to both lysyl-Sepharose and aminohexyl-Sepharose. Deleting the F domain (FGK1K2P versus GK1K2P and FGK2K1P versus GK2K1P) resulted in a 20-70% improvement of the interactions lysyl Sepharose and aminohexyl-Sepharose.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7479693 TI - Generation of analogs having potent antimicrobial and hemolytic activities with minimal changes from an inactive 16-residue peptide corresponding to the helical region of Staphylococcus aureus delta-toxin. AB - The delta-toxin is a 26-residue peptide from Staphylococcus aureus with the sequence formyl-MAQDIISTIGDLVKWIIDTVNKFTKK. NMR studies indicate that the segment IISTIGDLVKWIIDTV occurs in an alpha-helical conformation in the toxin. A synthetic peptide corresponding to this segment, although helical, did not exhibit hemolytic activity. Since charged residues like D and K are likely to modulate cytolytic activity, analogs of the 16-residue peptide were synthesized where D was systematically replaced by K. Analogs in which the first D and both Ds were replaced by K showed potent antimicrobial and hemolytic activities. The analog in which the second D was replaced by K was relatively less active. However, all the peptides showed an alpha-helical structure with similar helical content. The activities of the peptides were found to correlate directly with their ability to permeabilize model membranes. Thus, by minimal judicious replacement of charged amino acids, it should be possible to generate cytolytic peptides from short segments of peptide toxins. PMID- 7479692 TI - Engineering disulfide-linked single-chain Fv dimers [(sFv')2] with improved solution and targeting properties: anti-digoxin 26-10 (sFv')2 and anti-c-erbB-2 741F8 (sFv')2 made by protein folding and bonded through C-terminal cysteinyl peptides. AB - Single-chain Fv fusions with C-terminal cysteinyl peptides (sFv') have been engineered using model sFv proteins based upon the 26-10 anti-digoxin IgG and 741F8 anti-c-erbB-2 IgG monoclonal antibodies. As part of the 741F8 sFv construction process, the PCR-amplified 741F8 VH gene was modified in an effort to correct possible primer-induced errors. Genetic replacement of the N-terminal beta-strand sequence of 741F8 VH with that from the FR1 of anti-c-erbB-2 520C9 VH resulted in a dramatic improvement of sFv folding yields. Folding in urea glutathione redox buffers produced active sFv' with a protected C-terminal sulfhydryl, presumably as the mixed disulfide with glutathione. Disulfide-bonded (sFv')2 homodimers were made by disulfide interchange or oxidation after reductive elimination of the blocking group. Both 26-10 (sFv')2 and 741F8 (sFv')2 existed as stable dimers that were well behaved in solution, whereas 741F8 sFv and sFv' exhibited considerable self-association. The 741F8 sFv binds to the extracellular domain (ECD) of the c-erbB-2 oncogene protein, which is often overexpressed in breast cancer and other adenocarcinomas. The recombinant ECD was prepared to facilitate the analysis of 741F8 binding site properties; the cloned ECD gene, modified to encode a C-terminal Ser-Gly-His6 peptide, was transfected into Chinese hamster ovary cells using a vector that also expressed dihydrofolate reductase to facilitate methotrexate amplification. Optimized cell lines expressed ECD-His6 at high levels in a cell bioreactor; after isolation by immobilized metal affinity chromatography, final ECD yields were as high as 47 mg/l. An animal tumor model complemented physicochemical studies of 741F8 species and indicated increased tumor localization of the targeted 741F8 (sFv')2 over other monovalent 741F8 species. PMID- 7479694 TI - Prediction of the structure of GroES and its interaction with GroEL. AB - The three-dimensional structure of the GroES monomer and its interaction with GroEL has been predicted using a combination of prediction tools and experimental data obtained by biophysical [electron microscope (EM), Fourier transform infrared (FTIR), and nuclear magnetic resonance (NMR)] and biochemical techniques. The GroES monomer, according to the prediction, is composed of eight beta-strands forming a beta-barrel with loose ends. In the model, beta-strands 5 8 run along the outer surface of GroES, forming an antiparallel beta-sheet with beta 4 loosely bound to one of the edges. beta-strands 1-3 would then be parallel and placed in the interior of the molecule. Loops 1-3 would face the internal cavity of the GroEL-GroES complex, and together with conserved residues in loops 5 and 7, would form the active surface interacting with GroEL. PMID- 7479695 TI - Structural basis for serpin inhibitor activity. AB - The mechanism of formation and the structures of serpin-inhibitor complexes are not completely understood, despite detailed knowledge of the structures of a number of cleaved and uncleaved inhibitor, noninhibitor, and latent serpins. It has been proposed from comparison of inhibitor and noninhibitor serpins in the cleaved and uncleaved forms that insertion of strand s4A into preexisting beta sheet A is a requirement for serpin inhibitor activity. We have investigated the role of this strand in formation of serpin-proteinase complexes and in serpin inhibitor activity through homology modeling of wild type inhibitor, mutant substrate, and latent serpins, and of putative serpin-proteinase complexes. These models explain the high stability of the complexes and provide an understanding of substrate behavior in serpins with point mutations in s4A and of latency in plasminogen activator inhibitor I. PMID- 7479696 TI - Structural model of the photosynthetic reaction center of Rhodobacter capsulatus. AB - The reaction center (RC) from the photosynthetic bacterium Rhodobacter (Rb.) capsulatus has been the subject of a considerable amount of molecular biological and spectroscopic work aimed at improving our understanding of the primary steps of photosynthesis. However, no three-dimensional structure is available for this protein. We present here a model obtained by combining information from the structure of the highly homologous RC from Rhodopseudomonas (Rps.) viridis with molecular mechanics and simulated annealing calculations. In the Rb. capsulatus model the orientations of the bacteriochlorophyll monomer and the bacteriopheophytin on the branch inactive in electron transfer differ significantly from those in the RCs of Rps. viridis and Rb. sphaeroides. The bacteriopheophytin orientational difference is in good accord with previous linear dichroism measurements. A comparison is made of interactions between the pigments and the protein environment that may be of functional significance in Rps. viridis, Rb. sphaeroides, and Rb. capsulatus. PMID- 7479698 TI - The cytidylyltransferase superfamily: identification of the nucleotide-binding site and fold prediction. AB - The crystal structure of glycerol-3-phosphate cytidylyltransferase from B. subtilis (TagD) is about to be solved. Here, we report a testable structure prediction based on the identification by sequence analysis of a superfamily of functionally diverse but structurally similar nucleotide-binding enzymes. We predict that TagD is a member of this family. The most conserved region in this superfamily resembles the ATP-binding HiGH motif of class I aminoacyl-tRNA synthetases. The predicted secondary structure of cytidylyltransferase and its homologues is compatible with the alpha/beta topography of the class I aminoacyl tRNA synthetases. The hypothesis of similarity of fold is strengthened by sequence-structure alignment and 3D model building using the known structure of tyrosyl tRNA synthetase as template. The proposed 3D model of TagD is plausible both structurally, with a well packed hydrophobic core, and functionally, as the most conserved residues cluster around the putative nucleotide binding site. If correct, the model would imply a very ancient evolutionary link between class I tRNA synthetases and the novel cytidylyltransferase superfamily. PMID- 7479697 TI - The catalytic domain of a bacterial lytic transglycosylase defines a novel class of lysozymes. AB - The 70-kDa soluble lytic transglycosylase (SLT70) from Escherichia coli is a bacterial exo-muramidase that cleaves the cell wall peptidoglycan, producing 1,6 anhydro-muropeptides. The X-ray structure of SLT70 showed that one of its domains is structurally related to lysozyme, although there is no obvious similarity in amino acid sequence. To relate discrete structural features to differences in reaction mechanism and substrate/product specificity, we compared the three dimensional structure of the catalytic domain of SLT70 with the structures of three typical representatives of the lysozyme superfamily: chicken-type hen egg white lysozyme, goose-type swan egg-white lysozyme, and phage-type lysozyme from bacteriophage T4. We find a particularly close relationship between the catalytic domain of SLT70 and goose-type lysozyme, with not only a significant similarity in overall structure, but even a weak homology in amino acid sequence. This finding supports the notion that the goose-type lysozyme takes up a central position in the lysozyme superfamily and that it is structurally closest to the lysozyme ancestors. The saccharide-binding groove is the most conserved part in the four structures, but only two residues are absolutely preserved: the "catalytic" glutamic acid and a structurally required glycine. The "catalytic" aspartate is absent in SLT70, a difference that can be related to a different mechanism of cleavage of the beta-1,4-glycosidic bond. The unique composition of amino acids at the catalytic site, and the observation of a number of differences in the arrangements of secondary structure elements, define the catalytic domain of SLT70 as a novel class of lysozymes. Its fold is expected to be exemplary for other bacterial and bacteriophage muramidases with lytic transglycosylase activity. PMID- 7479699 TI - Hard alpha-keratin IF: a structural model lacking a head-to-tail molecular overlap but having hybrid features characteristic of both epidermal keratin and vimentin IF. AB - In intermediate filaments (IF) both epidermal keratin and vimentin molecules have been shown to have an eight residue head-to-tail overlap between the rod domains of similarly directed molecules. In the case of the epidermal keratins this region has also been shown to have particular structural/functional significance since it represents a hot-spot for mutations in the four keratinopathies characterized to date. While there is good evidence that this head-to-tail overlap is present in IF containing Type III, IV, and V chains, as well as in the epidermal keratin IF (Ib/IIb), there are no data currently available for the hard alpha-keratin IF (Ia/IIa). Using a variety of data derived from X-ray diffraction and crosslinking studies, as well as theoretical modeling, it is now possible to demonstrate that the overlap region is not a feature of hard alpha-keratin IF. Indeed, it is shown that there is a nine residue gap between consecutive parallel molecules in the IF. An explanation for this observation is presented in terms of compensating disulfide bonds that occur both within the IF, and between the IF and the matrix in which the IF are embedded. PMID- 7479700 TI - Size-independent comparison of protein three-dimensional structures. AB - Protein structures are routinely compared by their root-mean-square deviation (RMSD) in atomic coordinates after optimal rigid body superposition. What is not so clear is the significance of different RMSD values, particularly above the customary arbitrary cutoff for obvious similarity of 2-3 A. Our earlier work argued for an intrinsic cutoff for protein similarity that varied with the number of residues in the polypeptide chains being compared. Here we introduce a new measure, rho, of structural similarity based on RMSD that is independent of the sizes of the molecules involved, or of any other special properties of molecules. When rho is less than 0.4-0.5, protein structures are visually recognized to be obviously similar, but the mathematically pleasing intrinsic cutoff of rho < 1.0 corresponds to overall similarity in folding motif at a level not usually recognized until smoothing of the polypeptide chain path makes it striking. When the structures are scaled to unit radius of gyration and equal principle moments of inertia, the comparisons are even more universal, since they are no longer obscured by differences in overall size and ellipticity. With increasing chain length, the distribution of rho for pairs of random structures is skewed to higher values, but the value for the best 1% of the comparisons rises only slowly with the number of residues. This level is close to an intrinsic cutoff between similar and dissimilar comparisons, namely the maximal scaled rho possible for the two structures to be more similar to each other than one is to the other's mirror image. The intrinsic cutoff is independent of the number of residues or points being compared. For proteins having fewer than 100 residues, the 1% rho falls below the intrinsic cutoff, so that for very small proteins, geometrically significant similarity can often occur by chance. We believe these ideas will be helpful in judging success in NMR structure determination and protein folding modeling. PMID- 7479701 TI - Crystallization and preliminary crystallographic analysis of a 2,3 dihydroxybiphenyl dioxygenase from Pseudomonas sp. strain KKS102 having polychlorinated biphenyl (PCB)-degrading activity. AB - Crystals have been obtained for a 2,3-dihydroxybiphenyl dioxygenase (conventionally called BphC) from a polychlorinated biphenyl (PCB)-degrader, Pseudomonas sp. strain KKS102. The crystals were grown using both ammonium sulfate and MPD as the precipitating agents. The crystals belonged to a tetragonal space group (I422) and diffracted to 2.5 A. PMID- 7479702 TI - Cloning and expression of the uracil-DNA glycosylase inhibitor (UGI) from bacteriophage PBS-1 and crystallization of a uracil-DNA glycosylase-UGI complex. AB - The uracil-DNA glycosylase inhibitory protein (UGI) from the bacteriophage PBS-1 has been cloned and overexpressed. The nucleotide sequence is identical to that for the previously described PBS-2 inhibitor. The recombinant PBS-1 UGI inhibits the uracil-DNA glycosylase from herpes simplex virus type-1 (HSV-1 UDGase), and a complex between the HSV-1 UDGase and PBS-1 UGI has been crystallized. The crystals have unit cell dimensions a = 143.21 A, c = 40.78 A and are in a polar hexagonal space group. There is a single complex in the asymmetric unit with a solvent content of 62% by volume and the crystals diffract to 2.5A on a synchrotron radiation source. PMID- 7479703 TI - Crystallization and preliminary X-ray crystallographic studies of nonhistone region of macroH2A.1. AB - Histone macroH2A has a novel hybrid structure consisting of a large nonhistone region and a region that closely resembles a full-length histone H2A. One key to understanding macroH2A function is determining the structure and function of its nonhistone region. The nonhistone region of one of the two known macroH2A subtypes was expressed in Escherichia coli and purified using affinity and molecular sieve chromatography. Crystals of the protein suitable for structural studies were grown from polyethylene glycol solutions by vapor equilibration techniques. The crystals belong to the hexagonal space group P6(4) (or its enantiomorph P6(2)) with unit cell parameters: a = b = 106.2 A, c = 125.9 A, alpha = beta = 90 degrees, and gamma = 120 degrees. There are four molecules in the asymmetric unit. Self-rotation function studies revealed three twofold noncrystallographic rotation axes related approximately by 222 symmetry. These crystals have 47% solvent content and diffract to 3.8 A resolution. PMID- 7479704 TI - Crystallization and preliminary X-ray analysis of the cytoplasmic domain of human erythrocyte band 3. AB - A cytoplasmic domain of the human erythrocyte membrane protein band 3 (M(r) = 42,500), residues 1-379, expressed in and purified from E. coli, has been crystallized by the method of vapor diffusion in sitting drops with subsequent streak-seeding at room temperature. Initial crystals were grown from solutions containing 65-68% saturated ammonium sulfate at pH 4.9 and 2 mg/ml protein. Subsequent streak-seeding into solutions of 50-53% ammonium sulfate at pH 4.9 and 7 mg/ml protein produced single crystals suitable for X-ray analysis, which contained pure protein as revealed by gel electrophoresis. The crystals belong to the monoclinic space group C2 with cell dimensions of a = 178.8 A, b = 90.5 A, c = 122.1 A, and beta = 131.3 degrees and diffract at least to 2.7 A resolution (at 100 K). A self-rotation function shows the presence of approximate 222 local symmetry. PMID- 7479706 TI - Automatic identification of discrete substates in proteins: singular value decomposition analysis of time-averaged crystallographic refinements. AB - The singular value decomposition (SVD) provides a method for decomposing a molecular dynamics trajectory into fundamental modes of atomic motion. The right singular vectors are projections of the protein conformations onto these modes showing the protein motion in a generalized low-dimensional basis. Statistical analysis of the right singular vectors can be used to classify discrete configurational substates in the protein. The configuration space portraits formed from the right singular vectors can also be used to visualize complex high dimensional motion and to examine the extent of configuration space sampling by the simulation. PMID- 7479705 TI - A predicted consensus structure for the protein kinase C2 homology (C2H) domain, the repeating unit of synaptotagmin. AB - A secondary structure has been predicted for the protein kinase C2 regulatory domain found in homologous form in synaptotagmin, some phospholipases, and some GTP activated proteins. The proposed structure is built from seven consecutive beta strands followed by a terminal alpha helix. Considerations of overall surface exposure of individual secondary structural elements suggest that these are packed into a 2-sheet beta sandwich structure, with one of only three of the many possible folds being preferred. PMID- 7479707 TI - Phe-46(CD4) orients the distal histidine for hydrogen bonding to bound ligands in sperm whale myoglobin. AB - The role of Phe-46(CD4) in modulating the functional properties of sperm whale myoglobin was investigated by replacing this residue with Leu, Ile, Val, Ala, Trp, Tyr, and Glu. This highly conserved amino acid almost makes direct contact with the distal histidine and has been postulated to affect ligand binding. The overall association rate constants for CO, O2, and NO binding were little affected by decreasing the size of residue 46 step-wise from Phe to Leu to Val to Ala. In contrast, the rates of CO, O2, and NO dissociation increased 4-, 10-, and 25-fold, respectively, for the same series of mutants, causing large decreases in the affinity of myoglobin for all three diatomic gases. The rates of autooxidation at 37 degrees C, pH 7.0 increased dramatically from approximately 0.1-0.3 h-1 for wild-type, Tyr-46, and Trp-46 myoglobins to 1.5, 5.2, 4.9, and 5.0 h-1 for the Leu-46, Ile-46, Val-46 and Ala-46 mutants, respectively. Rates of NO and O2 geminate recombination were measured using 35 ps and 9 ns laser excitation pulses. Decreasing the size of residue 46 causes significant decreases in the extent of both picosecond and nanosecond rebinding processes. High resolution structures of Leu-46 and Val-46 metmyoglobins, Val-46 CO-myoglobin, and Val-46 deoxymyoglobin were determined by X-ray crystallography. When Phe-46 is replaced by Val, the loss of internal packing volume is compensated by (1) contraction of the CD corner toward the core of the protein, (2) movement of the E-helix toward the mutation site, (3) greater exposure of the distal pocket to intruding solvent molecules, and (4) large disorder in the position of the side chain of the distal histidine (His-64). In wild-type myoglobin, the van der Waals contact between C zeta of Phe-46 and C beta of His-64 appears to restrict rotation of the imidazole side chain. Insertion of Val at position 46 relieves this steric restriction, allowing the imidazole side chain to rotate about the C alpha - C beta bond toward the surface of the globin and about the C beta - C gamma bond toward the space previously occupied by the native Phe-46 side chain. This movement disrupts hydrogen bonding with bound ligands, causing significant decreases in affinity, and opens the distal pocket to solvent water molecules, causing marked increases in the rate of autooxidation.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 7479708 TI - Empirical correlation for the replacement of Ala by Gly: importance of amino acid secondary intrinsic propensities. AB - A series of Ala vs. Gly mutations at different helical and nonhelical positions of the chemotactic protein CheY, from E. coli, has been made. We have used this information to fit a general analytical equation that describes the free energy changes of an Ala to Gly mutation within +/- 0.45 kcal mol-1 with 95% confidence. The equation includes three terms: (1) the change in solvent-accessible hydrophobic surface area, corrected for the possible closure of the cavity left by deleting the C beta of the Ala; (2) the change in hydrophilic area of the nonintramolecularly hydrogen-bonded groups; and (3) the dihedral angles of the position being mutated. This last term extends the calculation to any conformation, not only alpha-helices. The general applicability of the equation for Ala vs. Gly mutations, when Ala or a small solvent-exposed polar residue is the wild-type residue, has been tested using data from other proteins: barnase, CI2 trypsin inhibitor, T4 lysozyme, and Staphylococcus nuclease. The predictive power of this simple approach offers the possibility of extending it to more complex mutations. PMID- 7479709 TI - Interaction stabilizing tertiary structure of bacteriorhodopsin studied by denaturation experiments. AB - The structural stability of bacteriorhodopsin was studied by denaturation experiments, using aliphatic alcohol as denaturants. The disappearance of a positive peak at 285 nm of the circular dichroism spectra, the change in the intrinsic fluorescence decay time, and the decrease of the regeneration activity bacteriorhodopsin indicated the denaturation of the tertiary structure of this protein at a methanol concentration of about 3 M. The circular dichroism band at 222 nm was unchanged by the denaturation. It was concluded that the alcohol denatured state in water was similar to the molten globule state of soluble proteins, in which only the tertiary structure was destroyed. Solvent substitution from water to hexane did not cause denaturation of bacteriorhodopsin. However, further addition of alcohol destroyed the secondary as well as the tertiary structures. Comparing the alcohol effects of bacteriorhodopsin in water to that in hexane, the dominant interactions for the structure formation of this protein could be revealed: the hydrophobic interaction that arose from the structure of water is essential for the stability of membrane spanning helices, while the interaction which binds the helices is polar in nature. PMID- 7479710 TI - A continuum theory for the prediction of lateral and rotational positioning of alpha-helices in membrane proteins: bacteriorhodopsin. AB - We have developed a new method for the prediction of the lateral and the rotational positioning of transmembrane helices, based upon the present status of knowledge about the dominant interaction of the tertiary structure formation. The basic assumption about the interaction is that the interhelix binding is due to the polar interactions and that very short extramembrane loop segments restrict the relative position of the helices. Another assumption is made for the simplification of the prediction that a helix may be regarded as a continuum rod having polar interaction fields around it. The polar interaction field is calculated by a probe helix method, using a copolymer of serine and alanine as probe helices. The lateral position of helices is determined by the strength of the interhelix binding estimated from the polar interaction field together with the length of linking loop segments. The rotational positioning is determined by the polar interaction field, assuming the optimum lateral configuration. The structural change due to the binding of a prosthetic group is calculated, fixing the rotational freedom of a helix that is connected to the prosthetic group. Applying this method to bacteriorhodopsin, the optimum lateral and rotational positioning of transmembrane helices that are very similar to the experimental configuration was obtained. This method was implemented by a software system, which was developed for this work, and automatic calculation became possible for membrane proteins comprised of several transmembrane helices. PMID- 7479711 TI - Multiple modes of ligand recognition: crystal structures of cyclin-dependent protein kinase 2 in complex with ATP and two inhibitors, olomoucine and isopentenyladenine. AB - Cyclin-dependent kinases (CDKs) are conserved regulators of the eukaryotic cell cycle with different isoforms controlling specific phases of the cell cycle. Mitogenic or growth inhibitory signals are mediated, respectively, by activation or inhibition of CDKs which phosphorylate proteins associated with the cell cycle. The central role of CDKs in cell cycle regulation makes them a potential new target for inhibitory molecules with anti-proliferative and/or anti neoplastic effects. We describe the crystal structures of the complexes of CDK2 with a weakly specific CDK inhibitor, N6-(delta 2-isopentenyl)adenine, and a strongly specific inhibitor, olomoucine. Both inhibitors are adenine derivatives and bind in the adenine binding pocket of CDK2, but in an unexpected and different orientation from the adenine of the authentic ligand ATP. The N6-benzyl substituent in olomoucine binds outside the conserved binding pocket and is most likely responsible for its specificity. The structural information from the CDK2 olomoucine complex will be useful in directing the search for the next generation inhibitors with improved properties. PMID- 7479712 TI - Effects of pH and high ionic strength on the adsorption and activity of native and mutated cellobiohydrolase I from Trichoderma reesei. AB - Cellobiohydrolase I (CBHI) is the major cellulase of Trichoderma reesei. The enzyme contains a discrete cellulose-binding domain (CBD), which increases its binding and activity on crystalline cellulose. We studied cellulase-cellulose interactions using site-directed mutagenesis on the basis of the three dimensional structure of the CBD of CBHI. Three mutant proteins which have earlier been produced in Saccharomyces cerevisiae were expressed in the native host organism. The data presented here support the hypothesis that a conserved tyrosine (Y492) located on the flat and more hydrophilic surface of the CBD is essential for the functionality. The data also suggest that the more hydrophobic surface is not directly involved in the CBD function. The pH dependence of the adsorption revealed that electrostatic repulsion between the bound proteins may also control the adsorption. The binding of CBHI to cellulose was significantly affected by high ionic strength suggesting that the interaction with cellulose includes a hydrophobic effect. High ionic strength increased the activity of the isolated core and of mutant proteins on crystalline cellulose, indicating that once productively bound, the enzymes are capable of solubilizing cellulose even with a mutagenized or with no CBD. PMID- 7479714 TI - Optimal local propensities for model proteins. AB - Lattice models of proteins were used to examine the role of local propensities in stabilizing the native state of a protein, using techniques drawn from spin-glass theory to characterize the free-energy landscapes. In the strong evolutionary limit, optimal conditions for folding are achieved when the contributions from local interactions to the stability of the native state is small. Further increasing the local interactions rapidly decreases the foldability. PMID- 7479713 TI - The heat capacity of proteins. AB - The heat capacity plays a major role in the determination of the energetics of protein folding and molecular recognition. As such, a better understanding of this thermodynamic parameter and its structural origin will provide new insights for the development of better molecular design strategies. In this paper we have analyzed the absolute heat capacity of proteins in different conformations. The results of these studies indicate that three major terms account for the absolute heat capacity of a protein: (1) one term that depends only on the primary or covalent structure of a protein and contains contributions from vibrational frequencies arising from the stretching and bending modes of each valence bond and internal rotations; (2) a term that contains the contributions of noncovalent interactions arising from secondary and tertiary structure; and (3) a term that contains the contributions of hydration. For a typical globular protein in solution the bulk of the heat capacity at 25 degrees C is given by the covalent structure term (close to 85% of the total). The hydration term contributes about 15 and 40% to the total heat capacity of the native and unfolded states, respectively. The contribution of non-covalent structure to the total heat capacity of the native state is positive but very small and does not amount to more than 3% at 25 degrees C. The change in heat capacity upon unfolding is primarily given by the increase in the hydration term (about 95%) and to a much lesser extent by the loss of noncovalent interactions (up to approximately 5%).(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7479715 TI - Activation of blood coagulation factor VIIa with cleaved tissue factor extracellular domain and crystallization of the active complex. AB - Exposure of blood to tissue factor leads to the formation of a high affinity tissue factor/factor VIIa complex which initiates blood coagulation. As a first step toward obtaining structural information of this enzyme system, a complex of active-site inhibited factor VIIa (F.VIIai) and soluble tissue factor (sTF) was prepared for crystallization. Crystals were obtained, but only after long incubation times. Analysis by SDS-PAGE and mass spectrometry indicated the presence of sTF fragments similar to those formed by proteolytic digestion with subtilisin (Konigsberg, W., Nemerson, Y., Fang, C., Lin, T.-C. Thromb. Haemost. 69:1171, 1993). To test the hypothesis that limited proteolysis of sTF facilitated the crystallization of the complex, sTF fragments were generated by subtilisin digestion and purified. Analysis by tandem mass spectrometry showed the presence of nonoverlapping N- and C-terminal sTF fragments encompassing more than 90% of the tissue factor extracellular domain. Enzymatic assays and binding studies demonstrated that an equimolar mixture of N- and C-terminal fragments bound to factor VIIa and fully restored cofactor activity. A complex of F.VIIai and sTF fragments was prepared for crystallization. Crystals were obtained using microseeding techniques. The best crystals had maximum dimensions of 0.12 x 0.12 x 0.6 mm and showed diffraction to a resolution of 3 A. PMID- 7479716 TI - Crystallization of Thermus thermophilus histidyl-tRNA synthetase and its complex with tRNAHis. AB - Histidyl-tRNA synthetase (HisRS) has been purified from the extreme thermophile Thermus thermophilus. The protein has been crystallized separately with histidine and with its cognate tRNAHis. Both crystals have been obtained using the vapor diffusion method with ammonium sulphate as precipitant. The crystals of HisRS with histidine belong to the spacegroup P2(1)2(1)2 with cell parameters a = 171.3 A, b = 214.7 A, c = 49.3 A, alpha = beta = gamma = 90 degrees. A complete data set to a resolution of 2.7A with an Rmerge on intensities of 4.1% has been collected on a single frozen crystal. A partial data set collected on a crystal of HisRS in complex with tRNAHis shows that the crystals are tetragonal with cell parameters a = b = 232 A, c = 559 A, alpha = beta = gamma = 90 degrees and diffract to about 4.5 A resolution. PMID- 7479717 TI - Commercial applications of speech interface technology: an industry at the threshold. AB - Speech interface technology, which includes automatic speech recognition, synthetic speech, and natural language processing, is beginning to have a significant impact on business and personal computer use. Today, powerful and inexpensive microprocessors and improved algorithms are driving commercial applications in computer command, consumer, data entry, speech-to-text, telephone, and voice verification. Robust speaker-independent recognition systems for command and navigation in personal computers are now available; telephone based transaction and database inquiry systems using both speech synthesis and recognition are coming into use. Large-vocabulary speech interface systems for document creation and read-aloud proofing are expanding beyond niche markets. Today's applications represent a small preview of a rich future for speech interface technology that will eventually replace keyboards with microphones and loud-speakers to give easy accessibility to increasingly intelligent machines. PMID- 7479718 TI - Military and government applications of human-machine communication by voice. AB - This paper describes a range of opportunities for military and government applications of human-machine communication by voice, based on visits and contacts with numerous user organizations in the United States. The applications include some that appear to be feasible by careful integration of current state of-the-art technology and others that will require a varying mix of advances in speech technology and in integration of the technology into applications environments. Applications that are described include (1) speech recognition and synthesis for mobile command and control; (2) speech processing for a portable multifunction soldier's computer; (3) speech- and language-based technology for naval combat team tactical training; (4) speech technology for command and control on a carrier flight deck; (5) control of auxiliary systems, and alert and warning generation, in fighter aircraft and helicopters; and (6) voice check-in, report entry, and communication for law enforcement agents or special forces. A phased approach for transfer of the technology into applications is advocated, where integration of applications systems is pursued in parallel with advanced research to meet future needs. PMID- 7479719 TI - Deployment of human-machine dialogue systems. AB - The deployment of systems for human-to-machine communication by voice requires overcoming a variety of obstacles that affect the speech-processing technologies. Problems encountered in the field might include variation in speaking style, acoustic noise, ambiguity of language, or confusion on the part of the speaker. The diversity of these practical problems encountered in the "real world" leads to the perceived gap between laboratory and "real-world" performance. To answer the question "What applications can speech technology support today?" the concept of the "degree of difficulty" of an application is introduced. The degree of difficulty depends not only on the demands placed on the speech recognition and speech synthesis technologies but also on the expectations of the user of the system. Experience has shown that deployment of effective speech communication systems requires an iterative process. This paper discusses general deployment principles, which are illustrated by several examples of human-machine communication systems. PMID- 7479720 TI - What does voice-processing technology support today? AB - This paper describes the state of the art in applications of voice-processing technologies. In the first part, technologies concerning the implementation of speech recognition and synthesis algorithms are described. Hardware technologies such as microprocessors and DSPs (digital signal processors) are discussed. Software development environment, which is a key technology in developing applications software, ranging from DSP software to support software also is described. In the second part, the state of the art of algorithms from the standpoint of applications is discussed. Several issues concerning evaluation of speech recognition/synthesis algorithms are covered, as well as issues concerning the robustness of algorithms in adverse conditions. PMID- 7479721 TI - User interfaces for voice applications. AB - This paper discusses some of the aspects of task requirements, user expectations, and technological capabilities that influence the design of a voice interface and then identifies several components of user interfaces that are particularly critical in successful voice applications. Examples from several applications are provided to demonstrate how these components are used to produce effective voice interfaces. PMID- 7479722 TI - Speech technology in the year 2001. AB - This paper introduces the session "Technology in the Year 2001" and is the first of four papers dealing with the future of human-machine communication by voice. In looking to the future it is important to recognize both the difficulties of technological forecasting and the frailties of the technology as it exists today- frailties that are manifestations of our limited scientific understanding of human cognition. The technology to realize truly advanced applications does not yet exist and cannot be supported by our presently incomplete science of speech. To achieve this long-term goal, the authors advocate a fundamental research program using a cybernetic approach substantially different from more conventional synthetic approaches. In a cybernetic approach, feedback control systems will allow a machine to adapt to a linguistically rich environment using reinforcement learning. PMID- 7479723 TI - Toward the ultimate synthesis/recognition system. AB - This paper predicts speech synthesis, speech recognition, and speaker recognition technology for the year 2001, and it describes the most important research problems to be solved in order to arrive at these ultimate synthesis and recognition systems. The problems for speech synthesis include natural and intelligible voice production, prosody control based on meaning, capability of controlling synthesized voice quality and choosing individual speaking style, multilingual and multidialectal synthesis, choice of application-oriented speaking styles, capability of adding emotion, and synthesis from concepts. The problems for speech recognition include robust recognition against speech variations, adaptation/normalization to variations due to environmental conditions and speakers, automatic knowledge acquisition for acoustic and linguistic modeling, spontaneous speech recognition, naturalness and ease of human-machine interaction, and recognition of emotion. The problems for speaker recognition are similar to those for speech recognition. The research topics related to all these techniques include the use of articulatory and perceptual constraints and evaluation methods for measuring the quality of technology and systems. PMID- 7479724 TI - Speech technology in 2001: new research directions. AB - Research in speech recognition and synthesis over the past several decades has brought speech technology to a point where it is being used in "real-world" applications. However, despite the progress, the perception remains that the current technology is not flexible enough to allow easy voice communication with machines. The focus of speech research is now on producing systems that are accurate and robust but that do not impose unnecessary constraints on the user. This chapter takes a critical look at the shortcomings of the current speech recognition and synthesis algorithms, discusses the technical challenges facing research, and examines the new directions that research in speech recognition and synthesis must take in order to form the basis of new solutions suitable for supporting a wide range of applications. PMID- 7479725 TI - New trends in natural language processing: statistical natural language processing. AB - The field of natural language processing (NLP) has seen a dramatic shift in both research direction and methodology in the past several years. In the past, most work in computational linguistics tended to focus on purely symbolic methods. Recently, more and more work is shifting toward hybrid methods that combine new empirical corpus-based methods, including the use of probabilistic and information-theoretic techniques, with traditional symbolic methods. This work is made possible by the recent availability of linguistic databases that add rich linguistic annotation to corpora of natural language text. Already, these methods have led to a dramatic improvement in the performance of a variety of NLP systems with similar improvement likely in the coming years. This paper focuses on these trends, surveying in particular three areas of recent progress: part-of-speech tagging, stochastic parsing, and lexical semantics. PMID- 7479726 TI - The future of voice-processing technology in the world of computers and communications. AB - This talk, which was the keynote address of the NAS Colloquium on Human-Machine Communication by Voice, discusses the past, present, and future of human-machine communications, especially speech recognition and speech synthesis. Progress in these technologies is reviewed in the context of the general progress in computer and communications technologies. PMID- 7479727 TI - Site-directed mutagenesis of Azotobacter vinelandii ferredoxin I: cysteine ligation of the [4Fe-4S] cluster with protein rearrangement is preferred over serine ligation. AB - The [4Fe-4S] cluster of Azotobacter vinelandii ferredoxin I receives three of its four ligands from a Cys-Xaa-Xaa-Cys-Xaa-Xaa-Cys sequence at positions 39-45 while the fourth ligand, Cys20, is provided by a distal portion of the sequence. Previously we reported that the site-directed mutation of Cys20 to Ala (C20A protein) resulted in the formation of a new [4Fe-4S] cluster that obtained its fourth ligand from Cys24, a free cysteine in the native structure. That ligand exchange required significant protein rearrangement. Here we report the conversion of Cys20 to Ser (C20S protein), which gives the protein the opportunity either to retain the native structure and use the Ser20 O gamma as a ligand or to rearrange and use Cys24. X-ray crystallography demonstrates that the cluster does not use the Ser20 O gamma as a ligand; rather it rearranges to use Cys24. In the C20S protein the [4Fe-4S] cluster has altered stability and redox properties relative to either C20A or the native protein. PMID- 7479729 TI - Direct structure analysis in protein electron crystallography: crystallographic phases for halorhodopsin to 6-A resolution. AB - The crystal structure of halorhodopsin was determined in (centrosymmetric) projection to 6-A resolution by direct methods that use only the amplitudes of the electron diffraction pattern. A multisolution technique was used to generate initial 15-A-resolution basis sets, and after selection of the best phase set (by the closest match of magnitude of Eobs and magnitude of Ecalc), annealing of individual reflections was used to improve its accuracy. The Sayre equation was then used to expand the phase terms to 10 A, followed again by phase annealing. A final expansion with the Sayre equation enlarged this corrected phase set to 6 A. When the condition of density flatness was used to locate the best phase solution after each extension, a final structure could be observed that was quite similar to the one found earlier by analysis of electron micrographs. PMID- 7479728 TI - Trans-dominant inhibitory human immunodeficiency virus type 1 protease monomers prevent protease activation and virion maturation. AB - Production of infectious human immunodeficiency virus (HIV) requires proper polyprotein processing by the dimeric viral protease. The trans-dominant inhibitory activity of a defective protease monomer with the active site Asp-25 changed to Asn was measured by transient transfection. A proviral plasmid that included the drug-selectable Escherichia coli gpt gene was used to deliver the wild-type (wt) or mutant proteases to cultured cells. Coexpression of the wt proviral DNA (HIV-gpt) with increasing amounts of the mutant proviral DNA (HIV gpt D25N) results in a concomitant decrease in proteolytic activity monitored by in vivo viral polyprotein processing. The viral particles resulting from inactivation of the protease were mostly immature, consisting predominantly of unprocessed p55gag and p160gag-pol polyproteins. In the presence of HIV-1 gp160 env, the number of secreted noninfectious particles correlated with the presence of increasing amounts of the defective protease. Greater than 97% reduction in infectivity was observed at a 1:6 ratio of wt to defective protease DNA. This provides an estimate of the level of inhibition required for effectively preventing virion processing. Stable expression of the defective protease in monkey cells reduced the yield of infectious particles from these cells by 90% upon transfection with the wt proviral DNA. These results show that defective subunits of the viral protease exert a trans-dominant inhibitory effect resulting from the formation of catalytically compromised heterodimers in vivo, ultimately yielding noninfectious viral particles. PMID- 7479730 TI - Lineage specification of neuronal precursors in the mouse spinal cord. AB - We have investigated the differentiation potential of precursor cells within the developing spinal cord of mice and have shown that spinal cord cells from embryonic day 10 specifically give rise to neurons when plated onto an astrocytic monolayer, Ast-1. These neurons had the morphology of motor neurons and > 83% expressed the motor neuron markers choline acetyltransferase, peripherin, calcitonin gene-related peptide, and L-14. By comparison, < 10% of the neurons arising on monolayers of other neural cell lines or 3T3 fibroblasts had motor neuron characteristics. Cells derived from dorsal, intermediate, and ventral regions of the spinal cord all behaved similarly and gave rise to motor neuron like cells when plated onto Ast-1. By using cells that expressed the lacZ reporter gene, it was shown that > 93% of cells present on the Ast-1 monolayers were motor neuron-like. Time-lapse analysis revealed that the precursors on the Ast-1 monolayers gave rise to neurons either directly or following a single cell division. Together, these results indicate that precursors in the murine spinal cord can be induced to differentiate into the motor neuron phenotype by factors produced by Ast-1 cells, suggesting that a similar factor(s) produced by cells akin to Ast-1 may regulate motor neuron differentiation in vivo. PMID- 7479731 TI - Nramp defines a family of membrane proteins. AB - Nramp (natural resistance-associated macrophage protein) is a newly identified family of integral membrane proteins whose biochemical function is unknown. We report on the identification of Nramp homologs from the fly Drosophila melanogaster, the plant Oryza sativa, and the yeast Saccharomyces cerevisiae. Optimal alignment of protein sequences required insertion of very few gaps and revealed remarkable sequence identity of 28% (yeast), 40% (plant), and 55% (fly) with the mammalian proteins (46%, 58%, and 73% similarity), as well as a common predicted transmembrane topology. This family is defined by a highly conserved hydrophobic core encoding 10 transmembrane segments. Other features of this hydrophobic core include several invariant charged residues, helical periodicity of sequence conservation suggesting conserved and nonconserved faces for several transmembrane helices, a consensus transport signature on the intracytoplasmic face of the membrane, and structural determinants previously described in ion channels. These characteristics suggest that the Nramp polypeptides form part of a group of transporters or channels that act on as yet unidentified substrates. PMID- 7479732 TI - Transcytosis of cholera toxin subunits across model human intestinal epithelia. AB - Cholera toxin (CT) elicits a massive secretory response from intestinal epithelia by binding apical receptors (ganglioside GM1) and ultimately activating basolateral effectors (adenylate cyclase). The mechanism of signal transduction from apical to basolateral membrane, however, remains undefined. We have previously shown that CT action on the polarized human intestinal epithelial cell line T84 requires endocytosis and processing in multiple intracellular compartments. Our aim in the present study was to test the hypothesis that CT may actually move to its site of action on the basolateral membrane by vesicular traffic. After binding apical receptors, CT entered basolaterally directed transcytotic vesicles. Both CT B subunits and to a lesser extent CT A subunits were delivered intact to the serosal surface of the basolateral membrane. The toxin did not traverse the monolayer by diffusion through intercellular junctions. Transcytosis of CT B subunits displayed nearly identical time course and temperature dependency with that of CT-induced Cl- secretion--suggesting the two may be related. These data identify a mechanism that may explain the link between the toxin's apical receptor and basolateral effector. PMID- 7479733 TI - Efficient gene transfer into human hepatocytes by baculovirus vectors. AB - Viral vectors are the most efficient tools for gene delivery, and the search for tissue-specific infecting viruses is important for the development of in vivo gene therapy strategies. The baculovirus Autographa californica nuclear polyhedrosis virus is widely used as a vector for expression of foreign genes in insect cells, and its host specificity is supposed to be restricted to arthropods. Here we demonstrate that recombinant A. californica nuclear polyhedrosis virus is efficiently taken up by human hepatocytes via an endosomal pathway. High-level reporter gene expression from heterologous promoters was observed in human and rabbit hepatocytes in vitro. Mouse hepatocytes and some other epithelial cell types are targeted at a considerably lower rate. The efficiency of gene transfer by baculovirus considerably exceeds that obtained by calcium phosphate or lipid transfection. These properties of baculovirus suggest a use for it as a vector for liver-directed gene transfer but highlight a potential risk in handling certain recombinant baculoviruses. PMID- 7479734 TI - A detergent-free method for purifying caveolae membrane from tissue culture cells. AB - Current methods for purifying caveolae from tissue culture cells take advantage of the Triton X-100 insolubility of this membrane domain. To circumvent the use of detergents, we have developed a method that depends upon the unique buoyant density of caveolae membrane. The caveolae fractions that we obtain are highly enriched in caveolin. As a consequence we are able to identify caveolae associated proteins that had previously gone undetected. Moreover, resident caveolae proteins that are soluble in Triton X-100 are retained during the isolation. PMID- 7479735 TI - Biosynthetic transport of the asialoglycoprotein receptor H1 to the cell surface occurs via endosomes. AB - Signals for endocytosis and for basolateral and lysosomal sorting are closely related in a number of membrane proteins, suggesting similar sorting mechanisms at the plasma membrane and in the trans-Golgi network (TGN). We tested the hypothesis that basolateral membrane proteins are transported to the cell surface via endosomes for the asialoglycoprotein receptor H1. This protein was tagged with a tyrosine sulfation site (H1TS) to allow specific labeling with [35S]sulfate in the TGN. Madin-Darby canine kidney cells expressing H1TS were pulse-labeled and chased for a period of time insufficient for labeled H1TS to reach the cell surface. Upon homogenization and gradient centrifugation, fractions devoid of TGN were subjected to immunoisolation of compartments containing mannose 6-phosphate receptor, which served as an endosomal marker. H1TS in transit to the cell surface was efficiently coisolated, whereas a labeled secretory protein and free glycosaminoglycan chains were not. This indicates an indirect pathway for the asialoglycoprotein receptor to the plasma membrane via endosomes and has important implications for protein sorting in the TGN and endosomes. PMID- 7479737 TI - Processing and cryopreservation of placental/umbilical cord blood for unrelated bone marrow reconstitution. AB - Clinical evidence of hematopoietic restoration with placental/umbilical cord blood (PCB) grafts indicates that PCB can be a useful source of hematopoietic stem cells for routine bone marrow reconstitution. In the unrelated setting, human leukocyte antigen (HLA)-matched donors must be obtained for candidate patients and, hence, large panels of frozen HLA-typed PCB units must be established. The large volume of unprocessed units, consisting mostly of red blood cells, plasma, and cryopreservation medium, poses a serious difficulty in this effort because storage space in liquid nitrogen is limited and costly. We report here that almost all the hematopoietic colony-forming cells present in PCB units can be recovered in a uniform volume of 20 ml by using rouleaux formation induced by hydroxyethyl starch and centrifugation to reduce the bulk of erythrocytes and plasma and, thus, concentrate leukocytes. This method multiples the number of units that can be stored in the same freezer space as much as 10 fold depending on the format of the storage system. We have also investigated the proportion of functional stem/progenitor cells initially present that are actually available to the recipient when thawed cryopreserved PCB units are infused. Progenitor cell viability is measurably decreased when thawed cells, still suspended in hypertonic cryopreservative solutions, are rapidly mixed with large volumes of isotonic solutions or plasma. The osmotic damage inflicted by the severe solute concentration gradient, however, can be averted by a simple 2 fold dilution after thawing, providing almost total recovery of viable hematopoietic progenitor cells. PMID- 7479736 TI - A Drosophila seminal fluid protein, Acp26Aa, stimulates egg laying in females for 1 day after mating. AB - Mating triggers behavioral and physiological changes in the Drosophila melanogaster female, including an elevation of egg laying. Seminal fluid molecules from the male accessory gland are responsible for initial behavioral changes, but persistence of these changes requires stored sperm. Using genetic analysis, we have identified a seminal fluid protein that is responsible for an initial elevation of egg laying. This molecule, Acp26Aa, has structural features of a prohormone and contains a region with amino acid similarity to the egg laying hormone of Aplysia. Acp26Aa is transferred to the female during mating, where it undergoes processing. Here we report the generation and analysis of mutants, including a null, in Acp26Aa. Females mated to male flies that lack Acp26Aa lay fewer eggs than do mates of normal males. This effect is apparent only on the first day after mating. The null mutation has no other detectable physiological or behavioral effects on the male or the mated female. PMID- 7479738 TI - Evidence for incorporation of intact dietary pyrimidine (but not purine) nucleosides into hepatic RNA. AB - The absorption and metabolism of dietary nucleic acids have received less attention than those of other organic nutrients, largely because of methodological difficulties. We supplemented the rations of poultry and mice with the edible alga Spirulina platensis, which had been uniformly labeled with 13C by hydroponic culture in 13CO2. The rations were ingested by a hen for 4 wk and by four mice for 6 days; two mice were fed a normal diet and two were fed a nucleic acid-deficient diet. The animals were killed and nucleosides were isolated from hepatic RNA. The isotopic enrichment of all mass isotopomers of the nucleosides was analyzed by selected ion monitoring of the negative chemical ionization mass spectrum and the labeling pattern was deconvoluted by reference to the enrichment pattern of the tracer material. We found a distinct difference in the 13C enrichment pattern between pyrimidine and purine nucleosides; the isotopic enrichment of uniformly labeled [M + 9] isotopomers of pyrimidines exceeded that of purines [M + 10] by > 2 orders of magnitude in the avian nucleic acids and by 7- and 14-fold in the murine nucleic acids. The purines were more enriched in lower mass isotopomers, those less than [M + 3], than the pyrimidines. Our results suggest that large quantities of dietary pyrimidine nucleosides and almost no dietary purine nucleosides are incorporated into hepatic nucleic acids without hydrolytic removal of the ribose moiety. In addition, our results support a potential nutritional role for nucleosides and suggest that pyrimidines are conditionally essential organic nutrients. PMID- 7479739 TI - Oxidative/reductive conjugation of mannan to antigen selects for T1 or T2 immune responses. AB - The induction of CD8+ cytotoxic T lymphocytes (CTLs) is desirable for immunization against many diseases, and recombinant-synthetic peptide antigens are now favored agents to use. However, a major problem is how to induce CTLs, which requires a T1-type response to such synthetic antigens. We report that T1 type (generating high CTL, low antibody) or T2-type (the reciprocal) responses can be induced by conjugation of the antigen to the carbohydrate polymer mannan: T1 responses are selected by using oxidizing conditions; T2 responses are selected by using reducing conditions for the conjugation. Using human MUC1 as a model antigen in mice, immunization with oxidized mannan-MUC1 fusion protein (ox M-FP) led to complete tumor protection (challenge up to 5 x 10(7) MUC1+ tumor cells), CTLs, and a high CTL precursor (CTLp) frequency (1/6900), whereas immunization with reduced mannan-MUC1 FP (red-M-FP) led to poor protection after challenge with only 10(6) MUC1+ tumor cells, no CTLs, and a low CTLp frequency (1/87,800). Ox-M-FP selects for a T1 response (mediated here by CD8+ cells) with high interferon gamma (IFN-gamma) secretion, no interleukin 4 (IL-4), and a predominant IgG2a antibody response; red-M-FP selects for a T2-type response with IL-4 production and a high predominant IgG1 antibody response but no IFN-gamma. PMID- 7479740 TI - High-affinity binding of Escherichia coli SecB to the signal sequence region of a presecretory protein. AB - The Escherichia coli cytosolic homotetrameric protein SecB is known to be involved in protein export across the plasma membrane. A currently prevalent view holds that SecB functions exclusively as a chaperone interacting nonspecifically with unfolded proteins, not necessarily exported proteins, whereas a contrary view holds that SecB functions primarily as a specific signal-recognition factor- i.e., in binding to the signal sequence region of exported proteins. To experimentally resolve these differences we assayed for binding between chemically pure SecB and chemically pure precursor (p) form (containing a signal sequence) and mature (m) form (lacking a signal sequence) of a model secretory protein (maltose binding protein, MBP) that was C-terminally truncated. Because of the C-terminal truncation, neither p nor m was able to fold. We found that SecB bound with 100-fold higher affinity to p (Kd 0.8 nM) than it bound to m (Kd 80 nM). As the presence of the signal sequence in p is the only feature that distinguished p from m, these data strongly suggest that the high-affinity binding of SecB is to the signal sequence region and not the mature region of p. Consistent with this conclusion, we found that a wild-type signal peptide, but not an export-incompetent mutant signal peptide of another exported protein (LamB), competed for binding to p. Moreover, the high-affinity binding of SecB to p was resistant to 1 M salt, whereas the low-affinity binding of SecB to m was not. These qualitative differences suggested that SecB binding to m was primarily by electrostatic interactions, whereas SecB binding to p was primarily via hydrophobic interactions, presumably with the hydrophobic core of the signal sequence. Taken together our data strongly support the notion that SecB is primarily a specific signal-recognition factor. PMID- 7479741 TI - The BCL2 major breakpoint region is a sequence- and cell-cycle-specific binding site of the Ku antigen. AB - The majority of translocations involving BCL2 are very narrowly targeted to three breakpoint clusters evenly spaced over a 100-bp region of the gene's terminal exon. We have recently shown that the immediate upstream boundary of this major breakpoint region (mbr) is a specific recognition site for single-strand DNA (ssDNA) binding proteins on the sense and antisense strands. The downstream flank of the mbr is a helicase binding site. In this report we demonstrate that the helicase and ssDNA binding proteins show reciprocal changes in binding activity over the cell cycle. The helicase is maximally active in G1 and early S phases; the ssDNA binding proteins are maximally active in late S and G2/M phases. An inhibitor of helicase binding appears in late S and G2/M. Finally, at least one component of the helicase binding complex is the Ku antigen. Thus, a protein with helicase activity implicated in repair of double-strand breaks, variable (diversity) joining recombination, and, potentially, cell-cycle regulation is targeted to the BCL2 mbr. PMID- 7479742 TI - Inhibition of phosphatidylinositol 3-kinase activity by association with 14-3-3 proteins in T cells. AB - Proteins of the 14-3-3 family can associate with, and/or modulate the activity of, several protooncogene and oncogene products and, thus, are implicated in regulation of signaling pathways. We report that 14-3-3 is associated with another important transducing enzyme, phosphatidylinositol 3-kinase (PI3-K). A recombinant 14-3-3 fusion protein bound several tyrosine-phosphorylated proteins from antigen receptor-stimulated T lymphocytes. PI3-K was identified by immunoblotting and enzymatic assays as one of the 14-3-3-binding proteins in resting or activated cells. Moreover, endogenous 14-3-3 and PI3-K were coimmunoprecipitated from intact T cells. Far-Western blots of gel-purified, immunoprecipitated PI3-K with a recombinant 14-3-3 fusion protein revealed direct binding of 14-3-3 to the catalytic subunit (p110) of PI3-K. Finally, anti phosphotyrosine immunoprecipitates from activated, 14-3-3-overexpressing cells contained lower PI3-K enzymatic activity than similar immunoprecipitates from control cells. These findings suggest that association of 14-3-3 with PI3-K in hematopoietic (and possibly other) cells regulates the enzymatic activity of PI3 K during receptor-initiated signal transduction. PMID- 7479743 TI - Mutagenesis of the human apolipoprotein B gene in a yeast artificial chromosome reveals the site of attachment for apolipoprotein(a). AB - Lipoprotein(a) [Lp(a)] is a lipoprotein formed by the disulfide linkage of apolipoprotein (apo) B100 of a low density lipoprotein particle to apolipoprotein(a). Prior studies have suggested that one of the C-terminal Cys residues of apo-B100 is involved in the disulfide linkage of apo-B100 to apo(a). To identify the apo-B100 Cys residue involved in the formation of Lp(a), we constructed a yeast artificial chromosome (YAC) spanning the human apo-B gene and used gene-targeting techniques to change Cys-4326 to Gly. The mutated YAC DNA was used to generate transgenic mice expressing the mutant human apo-B100 (Cys4326Gly). Unlike the wild-type human apo-B100, the mutant human apo-B100 completely lacked the ability to bind to apo(a) and form Lp(a). This study demonstrates that apo-B100 Cys-4326 is required for the assembly of Lp(a) and shows that gene targeting in YACs, followed by the generation of transgenic mice, is a useful approach for analyzing the structure of large proteins coded for by large genes. PMID- 7479744 TI - Changes in expression of putative antigens encoded by pigment genes in mouse melanomas at different stages of malignant progression. AB - Cutaneous melanomas of Tyr-SV40E transgenic mice (mice whose transgene consists of the tyrosinase promoter fused to the coding regions of simian virus 40 early genes) strikingly resemble human melanomas in their development and progression. Unlike human melanomas, the mouse tumors all arise in genetically identical individuals, thereby better enabling expression of specific genes to be characterized in relation to advancing malignancy. The products of pigment genes are of particular interest because peptides derived from these proteins have been reported to function as autoantigens with immunotherapeutic potential in some melanoma patients. However, the diminished pigmentation characteristic of many advanced melanomas raises the possibility that some of the relevant products may no longer be expressed in the most malignant cells. We have therefore investigated the contributions of several pigment genes in melanotic vs. relatively amelanotic components of primary and metastatic mouse melanomas. The analyses reveal marked differences within and among tumors in levels of mRNAs and proteins encoded by the wild-type alleles at the albino, brown, slaty, and silver loci. Tyrosinase (the protein encoded by the albino locus) was most often either absent or undetectable as melanization declined. The protein encoded by the slaty locus (tyrosinase-related protein 2) was the only one of those tested that was clearly present in all the tumor samples. These results suggest that sole reliance on targeting tyrosinase-based antigens might selectively favor survival of more malignant cells, whereas targeting the ensemble of the antigens tested might contribute toward a more inclusive and effective antimelanoma strategy. PMID- 7479745 TI - Agonist-induced desensitization of dopamine D1 receptor-stimulated adenylyl cyclase activity is temporally and biochemically separated from D1 receptor internalization. AB - The regulation of the dopamine D1 receptor was investigated by using c-myc epitope-tagged D1 receptors expressed in Sf9 (fall armyworm ovary) cells. Treatment of D1 receptors with 10 microM dopamine for 15 min led to a loss of the dopamine-detected high-affinity state of the receptor accompanying a 40% reduction in the ability of the receptor to mediate maximal dopamine stimulation of adenylyl cyclase activity. After 60 min of agonist exposure, 45 min after the occurrence of desensitization, 28% of the cell surface receptors were internalized into an intracellular light vesicular membrane fraction as determined by radioligand binding and supported by photoaffinity labeling, immunocytochemical staining, and immunoblot analysis. Pretreatment of cells with concanavalin A or sucrose completely blocked agonist-induced D1 receptor internalization without preventing agonist-induced desensitization, indicating a biochemical separation of these processes. Collectively, these findings indicate that the desensitization of D1 receptor-coupled adenylyl cyclase activity and D1 receptor internalization are temporarily and biochemically distinct mechanisms regulating D1 receptor function following agonist activation. PMID- 7479746 TI - A DNA sequencing strategy that requires only five bases of known terminal sequence for priming. AB - We have previously reported an enhanced version of sequencing by hybridization (SBH), termed positional SBH (PSBH). PSBH uses partially duplex probes containing single-stranded 3' overhangs, instead of simple single-stranded probes. Stacking interactions between the duplex probe and a single-stranded target allow us to reduce the probe sizes required to 5-base single-stranded overhangs. Here we demonstrate the use of PSBH to capture relatively long single-stranded DNA targets and perform standard solid-state Sanger sequencing on these primer template complexes without ligation. Our results indicate that only 5 bases of known terminal sequence are required for priming. In addition, the partially duplex probes have the ability to capture their specific target from a mixture of five single-stranded targets with different 3'-terminal sequences. This indicates the potential utility of the PSBH approach to sequence mixtures of DNA targets without prior purification. PMID- 7479747 TI - Long-term synaptic transformation of hippocampal CA1 gamma-aminobutyric acid synapses and the effect of anandamide. AB - Evidence is presented for a distinctive type of hippocampal synaptic modification [previously described for a molluscan gamma-aminobutyric acid (GABA) synapse after paired pre- and postsynaptic excitation]: transformation of GABA-mediated synaptic inhibition into synaptic excitation. This transformation persists with no further paired stimulation for 60 min or longer and is termed long-term transformation. Long-term transformation is shown to contribute to pairing induced long-term potentiation but not to long-term potentiation induced by presynaptic stimulation alone. Further support for such mechanistic divergence is provided by pharmacologic effects on long-term transformation as well as these two forms of long-term potentiation by Cl- channel blockers, glutamate and GABA antagonists, as well as the endogenous cannabinoid ligand anandamide. PMID- 7479748 TI - A cooperative model for receptor recognition and cell adhesion: evidence from the molecular packing in the 1.6-A crystal structure of the pheromone Er-1 from the ciliated protozoan Euplotes raikovi. AB - The crystal structure of the pheromone Er-1 from the unicellular eukaryotic organism Euplotes raikovi was determined at 1.6 A resolution and refined to a crystallographic R factor of 19.9%. In the tightly packed crystal, two extensive intermolecular helix-helix interactions arrange the Er-1 molecules into layers. Since the putative receptor of the pheromone is a membrane-bound protein, whose extracellular C-terminal domain is identical in amino acid sequence to the soluble pheromone, the interactions found in the crystal may mimic the pheromone receptor interactions as they occur on a cell surface. Based on this, we propose a model for the interaction between soluble pheromone molecules and their receptors. In this model, strong pheromone-receptor binding emerges as a consequence of the cooperative utilization of several weak interactions. The model offers an explanation for the results of binding studies and may also explain the adhesion between cells that occurs during mating. PMID- 7479749 TI - Mutations in the gene encoding the alpha subunit of the rod cGMP-gated channel in autosomal recessive retinitis pigmentosa. AB - Mutations in the genes encoding two proteins of the retinal rod phototransduction cascade, opsin and the beta subunit of rod cGMP phosphodiesterase, cause retinitis pigmentosa (RP) in some families. Here we report defects in a third member of this biochemical pathway in still other patients with this disease. We screened 94 unrelated patients with autosomal dominant RP and 173 unrelated patients with autosomal recessive RP for mutations in the gene encoding the alpha subunit of the rod cGMP-gated cation channel. Five mutant sequences cosegregated with disease among four unrelated families with autosomal recessive RP. Two of these were nonsense mutations early in the reading frame (Glu76End and Lys139End) and one was a deletion encompassing most if not all of the transcriptional unit; these three alleles would not be expected to encode a functional channel. The remaining two mutations were a missense mutation (Ser316Phe) and a frameshift [Arg654(1-bp del)] mutation truncating the last 32 aa in the C terminus. The latter two mutations were expressed in vitro and found to encode proteins that were predominantly retained inside the cell instead of being targeted to the plasma membrane. We conclude that the absence or paucity of functional cGMP-gated cation channels in the plasma membrane is deleterious to rod photoreceptors and is an uncommon cause of RP. PMID- 7479750 TI - Signaling by N- and C-terminal sequences of parathyroid hormone-related protein in hippocampal neurons. AB - Parathyroid hormone-related protein (PTHrP) is synthesized in the brain, and a single type of cloned receptor for the N-terminal portion of PTHrP and PTH is present in the central nervous system. Nothing is known about the physiological actions or signaling pathways used by PTHrP in the brain. Using cultured rat hippocampal neurons, we demonstrate that N-terminal PTHrP[1-34] and PTH[1-34] signal via cAMP and cytosolic calcium transients. The cAMP response showed strong acute (< or = 6 h) homologous and heterologous desensitization after preincubation with PTHrP or PTH. In contrast, the acute calcium response did not desensitize after preincubation with PTHrP; in fact, preincubation dramatically recruited additional responsive neurons. Unexpectedly, C-terminal PTHrP[107-139], which does not bind or activate the cloned PTH/PTHrP receptor, signaled in neurons via cytosolic calcium but not cAMP. Although some neurons responded to both PTHrP[1-34] and PTHrP[107-139], others responded only to PTHrP[1-34]. We conclude that certain hippocampal neurons exhibit dual signaling in response to PTHrP[1-34] and that some neurons have a receptor for C-terminal PTHrP that signals only via cytosolic calcium. PMID- 7479751 TI - Cloning and characterization of the Xenopus cyclin-dependent kinase inhibitor p27XIC1. AB - We have isolated a gene encoding Xic-1, a 27-kDa cyclin-dependent kinase (Cdk) inhibitor from Xenopus ovary that shares significant homology with both mammalian CIP1 and Kip1/Kip2. The N- and C-terminal halves of Xic-1 are sufficient for interacting with Cdks and proliferating cell nuclear antigen, respectively. Recombinant Xic-1 inhibits Xenopus cyclin E/Cdk2, cyclin A/Cdk2 and cyclin B/Cdc2 activities, although with quite different IC50 values. Truncation of the N terminus of Xic-1 increases the IC50 value for cyclin A/Cdk2 50-fold with no effect on the inhibition of cyclin E/Cdk2 or cyclin B/Cdc2.Xic-1 inhibits both single-stranded and nuclear DNA synthesis in egg extracts, an effect reversed by proliferating cell nuclear antigen or cyclin E/Cdk2, respectively. These results suggest a function for Xic-1 in the control of DNA synthesis by cyclin E/Cdk2. PMID- 7479752 TI - Transgenic mice carrying the diphtheria toxin A chain gene under the control of the granzyme A promoter: expected depletion of cytotoxic cells and unexpected depletion of CD8 T cells. AB - We have generated transgenic mice bearing the diphtheria toxin A chain (DTA) gene under the control of granzyme A (GrA) promoter sequences (GrA-DTA). GrA is expressed in activated cytotoxic cells but not in their immediate progenitors. These GrA-DTA mice are deficient in cytotoxic functions, indicating that most cytotoxic cells express GrA in vivo. Surprisingly, one founder strain containing a multicopy GrA-DTA insert show a marked and selective deficiency in CD8+ cells in peripheral lymphoid organs. This depletion was not observed in thymus, where the distribution of CD4+ and CD8+ cells is normal. Moreover, the emigration of T cells from thymus is normal, indicating that the depletion occurs in the periphery. GrA-DTA mice should be useful as models to dissect the role of cytotoxic cells in immune responses and as recipients of normal and neoplastic hematopoietic cells. The selective depletion of CD8+ cells in one founder strain could have implications for postthymic T-cell development. PMID- 7479753 TI - Mycoplasmas and oncogenesis: persistent infection and multistage malignant transformation. AB - Oncogenic potential of human mycoplasmas was studied using cultured mouse embryo cells, C3H/10T1/2 (C3H). Mycoplasma fermentans and Mycoplasma penetrans, mycoplasmas found in unusually high frequencies among patients with AIDS, were examined. Instead of acute transformation, a multistage process in promotion and progression of malignant cell transformation with long latency was noted; after 6 passages (1 wk per passage) of persistent infection with M. fermentans, C3H cells exhibited phenotypic changes with malignant characteristics that became progressively more prominent with further prolonged infection. Up to at least the 11th passage, all malignant changes were reversible if mycoplasmas were eradicated by antibiotic treatment. Further persistent infection with the mycoplasmas until 18 passages resulted in an irreversible form of transformation that included the ability to form tumors in animals and high soft agar cloning efficiency. Whereas chromosomal loss and translocational changes in C3H cells infected by either mycoplasma during the reversible stage were not prominent, the onset of the irreversible phase of transformation coincided with such karyotypic alteration. Genetic instability--i.e., prominent chromosomal alteration of permanently transformed cells--was most likely caused by mutation of a gene(s) responsible for fidelity of DNA replication or repair. Once induced, chromosomal alterations continued to accumulate both in cultured cells and in animals without the continued presence of the transforming microbes. Mycoplasma-mediated multistage oncogenesis exhibited here shares many characteristics found in the development of human cancer. PMID- 7479754 TI - Evidence for phosphatidylinositol 3-kinase as a regulator of endocytosis via activation of Rab5. AB - Phosphatidylinositol (PI) 3-kinases have been implicated in several aspects of intracellular membrane trafficking, although a detailed mechanism is yet to be established. In this study we demonstrated that wortmannin, a specific inhibitor of PI 3-kinases, inhibited constitutive endocytosis of horseradish peroxidase and transferrin in BHK-21 and TRVb-1 cells. The IC50 was approximately 40 ng/ml (93 nM). In addition, wortmannin blocked the stimulation of horseradish peroxidase uptake by the small GTPase Rab5 but not the stimulation by the GTPase-defective, constitutively activated Rab5 Gln79-->Leu mutant (Rab5:Q79L), providing further evidence that PI 3-kinase activity is essential for the early endocytic process. To further investigate the mechanism, we examined the effect of wortmannin on early endosome fusion in vitro. Wortmannin decreased endosome fusion by 80% with an IC50 value similar to that in intact cells. Addition of Rab5:Q79L but not wild type Rab5 reversed the inhibitory effect of wortmannin. Furthermore, addition of a constitutively activated PI 3-kinase but not its inactive counterpart stimulated early endosome fusion in vitro. These results strongly indicate that PI 3-kinase plays an important role in regulation of early endosome fusion, probably via activation of Rab5. PMID- 7479755 TI - Myc and Max: molecular evolution of a family of proto-oncogene products and their dimerization partner. AB - The myc gene family encodes a group of transcription factors that regulate cell proliferation and differentiation. These genes are widely studied because of their importance as proto-oncogenes. Phylogenetic analyses are described here for 45 Myc protein sequences representing c-, N-, L-, S-, and B-myc genes. A gene duplication early in vertebrate evolution produced the c-myc lineage and another lineage that later gave rise to the N- and L-myc lineages by another gene duplication. Evolutionary divergence in the myc gene family corresponds closely to the known branching order of the major vertebrate groups. The patterns of sequence evolution are described for five separate highly conserved regions, and these analyses show that differential rates of sequence divergence (= mosaic evolution) have occurred among conserved motifs. Further, the closely related dimerization partner protein Max exhibits significantly less sequence variability than Myc. It is suggested that the reduced variability in max stems from natural selection acting to preserve dimerization capability with products of myc and related genes. PMID- 7479756 TI - Subunit interactions in coordination of Ni2+ in cyclic nucleotide-gated channels. AB - Cyclic nucleotide-gated (CNG) channels present a unique model for studying the molecular mechanisms of channel gating. We have studied the mechanism of potentiation of expressed rod CNG channels by Ni2+ as a first step toward understanding the channel gating process. Here we report that coordination of Ni2+ between histidine residues (H420) on adjacent channel subunits occurs when the channels are open. Mutation of H420 to lysine completely eliminated the potentiation by Ni2+ but did not markedly alter the apparent cGMP affinity of the channel, indicating that the introduction of positive charge at the Ni(2+) binding site was not sufficient to produce potentiation. Deletion or mutation of most of the other histidines present in the channel did not diminish potentiation by Ni2+. We studied the role of subunit interactions in Ni2+ potentiation by generating heteromultimeric channels using tandem dimers of the rod CNG channel sequence. Injection of single heterodimers in which one subunit contained H420 and the other did not (wt/H420Q or H420Q/wt) resulted in channels that were not potentiated by Ni2+. However, coinjection of both heterodimers into Xenopus oocytes resulted in channels that exhibited potentiation. The H420 residues probably occurred predominantly in nonadjacent subunits when each heterodimer was injected individually, but, when the two heterodimers were coinjected, the H420 residues could occur in adjacent subunits as well. These results suggest that the mechanism of Ni2+ potentiation involves intersubunit coordination of Ni2+ by H420. Based on the preferential binding of Ni2+ to open channels, we suggest that alignment of H420 residues of neighboring subunits into the Ni(2+)-coordinating position may be associated with channel opening. PMID- 7479757 TI - Vector-mediated delivery of 125I-labeled beta-amyloid peptide A beta 1-40 through the blood-brain barrier and binding to Alzheimer disease amyloid of the A beta 1 40/vector complex. AB - The brain amyloid of Alzheimer disease (AD) may potentially be imaged in patients with AD by using neuroimaging technology and a radiolabeled form of the 40 residue beta-amyloid peptide A beta 1-40 that is enabled to undergo transport through the brain capillary endothelial wall, which makes up the blood-brain barrier (BBB) in vivo. Transport of 125I-labeled A beta 1-40 (125I-A beta 1-40) through the BBB was found to be negligible by experiments with both an intravenous injection technique and an internal carotid artery perfusion method in anesthetized rats. In addition, 125I-A beta 1-40 was rapidly metabolized after either intravenous injection or internal carotid artery perfusion. BBB transport was increased and peripheral metabolism was decreased by conjugation of monobiotinylated 125I-A beta 1-40 to a vector-mediated drug delivery system, which consisted of a conjugate of streptavidin (SA) and the OX26 monoclonal antibody to the rat transferrin receptor, which undergoes receptor-mediated transcytosis through the BBB. The brain uptake, expressed as percent of injected dose delivered per gram of brain, of the 125I,bio-A beta 1-40/SA-OX26 conjugate was 0.15 +/- 0.01, a level that is 2-fold greater than the brain uptake of morphine. The binding of the 125I,bio-A beta 1-40/SA-OX26 conjugate to the amyloid of AD brain was demonstrated by both film and emulsion autoradiography performed on frozen sections of AD brain. Binding of the 125I,bio-A beta 1-40/SA OX26 conjugate to the amyloid of AD brain was completely inhibited by high concentrations of unlabeled A beta 1-40. In conclusion, these studies show that BBB transport and access to amyloid within brain may be achieved by conjugation of A beta 1-40 to a vector-mediated BBB drug delivery system. PMID- 7479759 TI - Light intensity regulation of cab gene transcription is signaled by the redox state of the plastoquinone pool. AB - The eukaryotic green alga Dunaliella tertiolecta acclimates to decreased growth irradiance by increasing cellular levels of light-harvesting chlorophyll protein complex apoproteins associated with photosystem II (LHCIIs), whereas increased growth irradiance elicits the opposite response. Nuclear run-on transcription assays and measurements of cab mRNA stability established that light intensity dependent changes in LHCII are controlled at the level of transcription. cab gene transcription in high-intensity light was partially enhanced by reducing plastoquinone with 3-(3,4-dichlorophenyl)-1,1-dimethyl urea (DCMU), whereas it was repressed in low-intensity light by partially inhibiting the oxidation of plastoquinol with 2,5-dibromo-3-methyl-6-isopropyl-p-benzoquinone (DBMIB). Uncouplers of photosynthetic electron transport and inhibition of water splitting had no effect on LHCII levels. These results strongly implicate the redox state of the plastoquinone pool in the chloroplast as a photon-sensing system that is coupled to the light-intensity regulation of nuclear-encoded cab gene transcription. The accumulation of cellular chlorophyll at low-intensity light can be blocked with cytoplasmically directed phosphatase inhibitors, such as okadaic acid, microcystin L-R, and tautomycin. Gel mobility-shift assays revealed that cells grown in high-intensity light contained proteins that bind to the promoter region of a cab gene carrying sequences homologous to higher plant light responsive elements. On the basis of these experimental results, we propose a model for a light intensity signaling system where cab gene expression is reversibly repressed by a phosphorylated factor coupled to the redox status of plastoquinone through a chloroplast protein kinase. PMID- 7479758 TI - Crystal structure of the cell cycle-regulatory protein suc1 reveals a beta-hinge conformational switch. AB - The Schizosaccharomyces pombe cell cycle-regulatory protein suc1, named as the suppressor of cdc2 temperature-sensitive mutations, is essential for cell cycle progression. To understand suc1 structure-function relationships and to help resolve conflicting interpretations of suc1 function based on genetic studies of suc1 and its functional homologs in both lower and higher eukaryotes, we have determined the crystal structure of the beta-interchanged suc1 dimer. Each domain consists of three alpha-helices and a four-stranded beta-sheet, completed by the interchange of terminal beta-strands between the two subunits. This beta interchanged suc1 dimer, when compared with the beta-hairpin single-domain folds of suc1, reveals a beta-hinge motif formed by the conserved amino acid sequence HVPEPH. This beta-hinge mediates the subunit conformation and assembly of suc1: closing produces the intrasubunit beta-hairpin and single-domain fold, whereas opening leads to the intersubunit beta-strand interchange and interlocked dimer assembly reported here. This conformational switch markedly changes the surface accessibility of sequence-conserved residues available for recognition of cyclin dependent kinase, suggesting a structural mechanism for beta-hinge-mediated regulation of suc1 biological function. Thus, suc1 belongs to the family of domain-swapping proteins, consisting of intertwined and dimeric protein structures in which the dual assembly modes regulate their function. PMID- 7479760 TI - Interactions of a Rel protein with its inhibitor. AB - Cactus, a Drosophila homologue of I kappa B, binds to and inhibits Dorsal, a homologue of the p50 and p65 components of NF-kappa B. We describe experiments in yeast with various Dorsal and Cactus derivatives showing that Cactus blocks the DNA binding and nuclear localization functions of Dorsal. In contrast, Dorsal's transcriptional activating region is functional in the Dorsal-Cactus complex. We identify two Dorsal mutants, Dorsal C233R and Dorsal S234P, that escape Cactus inhibition in vivo, and we show that these mutants fail to interact with Cactus in vitro. From this and data of others, we identify the likely surface of Dorsal that binds Cactus. We also describe a modified PCR mutagenesis procedure, easier to use than conventional methods, that produces a library of high complexity. PMID- 7479761 TI - The inability of phosphatidylinositol 3-kinase activation to stimulate GLUT4 translocation indicates additional signaling pathways are required for insulin stimulated glucose uptake. AB - Recent experimental evidence has focused attention to the role of two molecules, insulin receptor substrate 1 (IRS-1) and phosphatidylinositol 3-kinase (PI3 kinase), in linking the insulin receptor to glucose uptake; IRS-1 knockout mice are insulin resistant, and pharmacological inhibitors of PI3-kinase block insulin stimulated glucose uptake. To investigate the role of PI3-kinase and IRS-1 in insulin-stimulated glucose uptake we examined whether stimulation of insulin sensitive cells with platelet-derived growth factor (PDGF) or with interleukin 4 (IL-4) stimulates glucose uptake; the activated PDGF receptor (PDGFR) directly binds and activates PI3-kinase, whereas the IL-4 receptor (IL-4R) activates PI3 kinase via IRS-1 or the IRS-1-related molecule 4PS. We found that stimulation of 3T3-L1 adipocytes with PDGF resulted in tyrosine phosphorylation of the PDGFR and activation of PI3-kinase in these cells. To examine whether IL-4 stimulates glucose uptake, L6 myoblasts were engineered to overexpress GLUT4 as well as both chains of the IL-4R (L6/IL-4R/GLUT4); when these L6/IL-4R/GLUT4 myoblasts were stimulated with IL-4, IRS-1 became tyrosine phosphorylated and associated with PI3-kinase. Although PDGF and IL-4 can activate PI3-kinase in the respective cell lines, they do not possess insulin's ability to stimulate glucose uptake and GLUT4 translocation to the plasma membrane. These findings indicate that activation of PI3-kinase is not sufficient to stimulate GLUT4 translocation to the plasma membrane. We postulate that activation of a second signaling pathway by insulin, distinct from PI3-kinase, is necessary for the stimulation of glucose uptake in insulin-sensitive cells. PMID- 7479762 TI - Contraction due to microtubule disruption is associated with increased phosphorylation of myosin regulatory light chain. AB - Microtubules have been proposed to function as rigid struts which oppose cellular contraction. Consistent with this hypothesis, microtubule disruption strengthens the contractile force exerted by many cell types. We have investigated alternative explanation for the mechanical effects of microtubule disruption: that microtubules modulate the mechanochemical activity of myosin by influencing phosphorylation of the myosin regulatory light chain (LC20). We measured the force produced by a population of fibroblasts within a collagen lattice attached to an isometric force transducer. Treatment of cells with nocodazole, an inhibitor of microtubule polymerization, stimulated an isometric contraction that reached its peak level within 30 min and was typically 30-45% of the force increase following maximal stimulation with 30% fetal bovine serum. The contraction following nocodazole treatment was associated with a 2- to 4-fold increase in LC20 phosphorylation. The increases in both force and LC20 phosphorylation, after addition of nocodazole, could be blocked or reversed by stabilizing the microtubules with paclitaxel (former generic name, taxol). Increasing force and LC20 phosphorylation by pretreatment with fetal bovine serum decreased the subsequent additional contraction upon microtubule disruption, a finding that appears inconsistent with a load-shifting mechanism. Our results suggest that phosphorylation of LC20 is a common mechanism for the contractions stimulated both by microtubule poisons and receptor-mediated agonists. The modulation of myosin activity by alterations in microtubule assembly may coordinate the physiological functions of these cytoskeletal components. PMID- 7479763 TI - Proteolysis of major histocompatibility complex class II-associated invariant chain is regulated by the alternatively spliced gene product, p41. AB - Invariant chain (Ii) is an intracellular type II transmembrane glycoprotein that is associated with major histocompatibility complex class II molecules during biosynthesis. Ii exists in two alternatively spliced forms, p31 and p41. Both p31 and p41 facilitate folding of class II molecules, promote egress from the endoplasmic reticulum, prevent premature peptide binding, and enhance localization to proteolytic endosomal compartments that are thought to be the sites for Ii degradation, antigen processing, and class II-peptide association. In spite of the dramatic and apparently equivalent effects that p31 and p41 have on class II biosynthesis, the ability of invariant chain to enhance antigen presentation to T cells is mostly restricted to p41. Here we show that degradation of Ii leads to the generation of a 12-kDa amino-terminal fragment that in p41-positive, but not in p31-positive, cells remains associated with class II molecules for an extended time. Interestingly, we find that coexpression of the two isoforms results in a change in the pattern of p31 degradation such that endosomal processing of p31 also leads to extended association of a similar 12-kDa fragment with class II molecules. These data raise the possibility that p41 may have the ability to impart its pattern of proteolytic processing on p31 molecules expressed in the same cells. This would enable a small number of p41 molecules to modify the post-translational transport and/or processing of an entire cohort of class II-Ii complexes in a manner that could account for the unique ability of p41 to enhance antigen presentation. PMID- 7479764 TI - Identification of haptoglobin as a natural inhibitor of trypanocidal activity in human serum. AB - Trypanosomes are protozoan parasites of medical and veterinary importance. Trypanosoma brucei rhodesiense and Trypanosoma brucei gambiense infect humans, causing African sleeping sickness. However, Trypanosoma brucei brucei can only infect animals, causing the disease Nagana in cattle. Man is protected from this subspecies of trypanosomes by a toxic subtype of high density lipoproteins (HDLs) called the trypanosome lytic factor (TLF). The toxic molecule in TLF is believed to be the haptoglobin-related protein that when bound to hemoglobin kills the trypanosome via oxidative damage initiated by its peroxidase activity. The amount of lytic activity in serum varies widely between different individuals with up to a 60-fold difference in activity. In addition, an increase in the total amount of lytic activity occurs during the purification of TLF, suggesting that an inhibitor of TLF (ITLF) exists in human serum. We now show that the individual variation in trypanosome lytic activity in serum correlates to variations in the amount of ITLF. Immunoblots of ITLF probed with antiserum against haptoglobin recognize a 120-kDa protein, indicating that haptoglobin is present in partially purified ITLF. Haptoglobin involvement is further shown in that it inhibits TLF in a manner similar to ITLF. Using an anti-haptoglobin column to remove haptoglobin from ITLF, we show that the loss of haptoglobin coincides with the loss of inhibitor activity. Addition of purified haptoglobin restores inhibitor activity. This indicates that haptoglobin is the molecule responsible for inhibition and therefore causing the individual variation in serum lytic activity. PMID- 7479766 TI - Calcium signaling in a narrow somatic submembrane shell during synaptic activity in cerebellar Purkinje neurons. AB - Temporal and spatial changes in the intracellular Ca2+ concentration ([Ca2+]i) were examined in dendrites and somata of rat cerebellar Purkinje neurons by combining whole-cell patch-clamp recording and fast confocal laser-scanning microscopy. In cells loaded via the patch pipette with the high-affinity Ca2+ indicator Calcium Green-1 (Kd approximately 220 nM), a single synaptic climbing fiber response, a so-called complex spike, resulted in a transient elevation of [Ca2+]i that showed distinct differences among various subcellular compartments. With conventional imaging, the Ca2+ signals were prominent in the dendrites and almost absent in the soma. Confocal recordings from the somatic region, however, revealed steep transient increases in [Ca2+]i that were confined to a submembrane shell of 2- to 3-microns thickness. In the central parts of the soma [Ca2+]i increases were much slower and had smaller amplitudes. The kinetics and amplitudes of the changes in [Ca2+]i were analyzed in more detail by using the fast, low-affinity Ca2+ indicator Calcium Green-5N (Kd approximately 17 microM). We found that brief depolarizing pulses produced [Ca2+]i increases in a narrow somatic submembrane shell that resembled those seen in the dendrites. These results provide direct experimental evidence that the surface-to-volume ratio is a critical determinant of the spatiotemporal pattern of Ca2+ signals evoked by synaptic activity in neurons. PMID- 7479765 TI - Development of a safe live Leishmania vaccine line by gene replacement. AB - Vaccination with live Leishmania major has been shown to yield effective immunization in humans; however, this has been discontinued because of problems associated with virulence of the available vaccine lines. To circumvent this, we tested the ability of a dhfr-ts- null mutant of L. major, obtained by gene targeting, to infect and then to vaccinate mice against challenge with virulent L. major. Survival and replication of dhfr-ts- in macrophages in vitro were dependent upon thymidine, with parasites differentiating into amastigotes prior to destruction. dhfr-ts- parasites persisted in BALB/c mice for up to 2 months, declining with a half-life of 2-3 days. Nonetheless, dhfr-ts- was incapable of causing disease in both susceptible and immunodeficient (nu/nu) BALB/c mice. Animal infectivity could be partially restored by thymidine supplementation. When inoculated by the i.v., s.c., or i.m. routes into mice, dhfr-ts- could elicit substantial resistance to a subsequent challenge with virulent L. major. Thus, Leishmania bearing auxotrophic gene knockouts can be safe and induce protective immunity. Potentially, dhfr-ts- could be used as a platform for delivery of immunogens relevant to other diseases. PMID- 7479768 TI - Abr and Bcr are multifunctional regulators of the Rho GTP-binding protein family. AB - Philadelphia chromosome-positive leukemias result from the fusion of the BCR and ABL genes, which generates a functional chimeric molecule. The Abr protein is very similar to Bcr but lacks a structural domain which may influence its biological regulatory capabilities. Both Abr and Bcr have a GTPase-activating protein (GAP) domain similar to those found in other proteins that stimulate GTP hydrolysis by members of the Rho family of GTP-binding proteins, as well as a region of homology with the guanine nucleotide dissociation-stimulating domain of the DBL oncogene product. We purified as recombinant fusion proteins the GAP- and Dbl-homology domains of both Abr and Bcr. The Dbl-homology domains of Bcr and Abr were active in stimulating GTP binding to CDC42Hs, RhoA, Rac1, and Rac2 (rank order, CDC42Hs > RhoA > Rac1 = Rac2) but were inactive toward Rap1A and Ha-Ras. Both Bcr and Abr acted as GAPs for Rac1, Rac2, and CDC42Hs but were inactive toward RhoA, Rap1A, and Ha-Ras. Each individual domain bound in a noncompetitive manner to GTP-binding protein substrates. These data suggest the multifunctional Bcr and Abr proteins might interact simultaneously and/or sequentially with members of the Rho family to regulate and coordinate cellular signaling. PMID- 7479767 TI - Crystal structure of the I-domain from the CD11a/CD18 (LFA-1, alpha L beta 2) integrin. AB - We report the 1.8-A crystal structure of the CD11a I-domain with bound manganese ion. The CD11a I-domain contains binding sites for intercellular adhesion molecules 1 and 3 and can exist in both low- and high-affinity states. The metal bound form reported here is likely to represent a high-affinity state. The CD11a I-domain structure reveals a strained hydrophobic ridge adjacent to the bound metal ion that may serve as a ligand-binding surface and is likely to rearrange in the absence of bound metal ion. The CD11a I-domain is homologous to domains found in von Willebrand factor, and mapping of mutations found in types 2a and 2b von Willebrand disease onto this structure allows consideration of the molecular basis of these forms of the disease. PMID- 7479770 TI - Detection of membrane-bound HLA-G translated products with a specific monoclonal antibody. AB - A monomorphic anti-HLA-G monoclonal antibody (mAb) was obtained by immunization of HLA-B27/human beta 2-microglobulin double-transgenic mice with transfected murine L cells expressing both HLA-G and human beta 2-microglobulin. This mAb, designated BFL.1, specifically recognizes, by flow cytometry analysis, the immunizing HLA-G-expressing cells, whereas it does not bind to parental untransfected or to HLA-B7- and HLA-A3-transfected L cells, suggesting that it distinguishes between classical HLA-A and -B and nonclassical HLA-G class I molecules. This was further assessed by the absence of BFL.1 reactivity with a number of human cell lines known to express classical HLA class I proteins. In addition, we showed that the BFL.1 mAb also labels HLA-G-naturally-expressing JEG 3 and HLA-G-transfected JAR human choriocarcinoma cell lines as well as a subpopulation of first-trimester placental cytotrophoblast cells. Further biochemical studies were performed by immunoprecipitation of biotinylated membrane lysates: BFL.1, like the monomorphic W6/32 mAb, immunoprecipitated a 39 kDa protein in HLA-G-expressing cell lines, a size corresponding to the predicted full-length HLA-G1 isoform. However, in contrast to W6/32, which immunoprecipitates both classical and nonclassical HLA class I heavy chains, BFL.1 mAb does not recognize the class Ia products. Such a mAb should be a useful tool for analysis of HLA-G protein expression in various normal and pathological human tissues and for determination of the function(s) of translated HLA-G products. PMID- 7479769 TI - Grb-IR: a SH2-domain-containing protein that binds to the insulin receptor and inhibits its function. AB - To identify potential signaling molecules involved in mediating insulin-induced biological responses, a yeast two-hybrid screen was performed with the cytoplasmic domain of the human insulin receptor (IR) as bait to trap high affinity interacting proteins encoded by human liver or HeLa cDNA libraries. A SH2-domain-containing protein was identified that binds with high affinity in vitro to the autophosphorylated IR. The mRNA for this protein was found by Northern blot analyses to be highest in skeletal muscle and was also detected in fat by PCR. To study the role of this protein in insulin signaling, a full-length cDNA encoding this protein (called Grb-IR) was isolated and stably expressed in Chinese hamster ovary cells overexpressing the human IR. Insulin treatment of these cells resulted in the in situ formation of a complex of the IR and the 60 kDa Grb-IR. Although almost 75% of the Grb-IR protein was bound to the IR, it was only weakly tyrosine-phosphorylated. The formation of this complex appeared to inhibit the insulin-induced increase in tyrosine phosphorylation of two endogenous substrates, a 60-kDa GTPase-activating-protein-associated protein and, to a lesser extent, IR substrate 1. The subsequent association of this latter protein with phosphatidylinositol 3-kinase also appeared to be inhibited. These findings raise the possibility that Grb-IR is a SH2-domain-containing protein that directly complexes with the IR and serves to inhibit signaling or redirect the IR signaling pathway. PMID- 7479771 TI - Detection of Ca2+ entry through mechanosensitive channels localizes the site of mechanoelectrical transduction in hair cells. AB - A hair cell, the sensory receptor of the internal ear, transduces mechanical stimuli into electrical responses. Transduction results from displacement of the hair bundle, a cluster of rod-shaped stereocilia extending from the cell's apical surface. Biophysical experiments indicate that, by producing shear between abutting stereocilia, a bundle displacement directly opens cation-selective transduction channels. Specific models of gating depend on the location of these channels, which has been controversial: although some physiological and immunocytochemical experiments have situated the transduction channels at the hair bundle's top, monitoring of fluorescence signals from the Ca2+ indicator fura-2 has instead suggested that Ca2+ traverses channels at the bundle's base. To examine the site of Ca2+ entry through transduction channels, we used laser scanning confocal microscopy, with a spatial resolution of < 1 micron and a temporal resolution of < 2 ms, to observe hair cells filled with the indicator fluo-3. An unstimulated hair cell showed a "tip blush" of enhanced fluorescence at the hair bundle's top, which we attribute to Ca2+ permeation through transduction channels open at rest. Upon mechanical stimulation, individual stereocilia displayed increased fluorescence that originated near their tips, then spread toward their bases. Our results confirm that mechanoelectrical transduction occurs near stereociliary tips. PMID- 7479772 TI - The purification and characterization of fetal liver hematopoietic stem cells. AB - Thy-1loSca-1+Lin-Mac-1+CD4- cells have been isolated from the livers of C57BL-Thy 1.1 fetuses. This population appears to be an essentially pure population of hematopoietic stem cells (HSC), in that injection of only six cells into lethally irradiated adult recipients yields a limit dilution frequency of donor cell reconstituted mice. Sixty-seven to 77% of clones in this population exhibit long term multilineage progenitor activity. This population appears to include all long-term multilineage reconstituting progenitors in the fetal liver. A high proportion of cells are in cycle, and the absolute number of cells in this population doubles daily in the fetal liver until 14.5 days postcoitum. At 15.5 days postcoitum, the frequency of this population falls dramatically. Long-term reconstituting HSC clones from the fetal liver give rise to higher levels of reconstitution in lethally irradiated mice than long-term reconstituting HSC from the bone marrow. The precise phenotypic and functional characteristics of HSC vary according to tissue and time during ontogeny. PMID- 7479773 TI - Opposing early and late effects of insulin-like growth factor I on differentiation and the cell cycle regulatory retinoblastoma protein in skeletal myoblasts. AB - The mechanisms by which insulin-like growth factors (IGFs) can be both mitogenic and differentiation-promoting in skeletal myoblasts are unclear because these two processes are believed to be mutually exclusive in this tissue. The phosphorylation state of the ubiquitous nuclear retinoblastoma protein (Rb) plays an important role in determining whether myoblasts proliferate or differentiate: Phosphorylated Rb promotes mitogenesis, whereas un- (or hypo-) phosphorylated Rb promotes cell cycle exit and differentiation. We hypothesized that IGFs might affect the fate of myoblasts by regulating the phosphorylation of Rb. Although long-term IGF treatment is known to stimulate differentiation, we find that IGFs act initially to inhibit differentiation and are exclusively mitogenic. These early effects of IGFs are associated with maintenance of Rb phosphorylation typical of proliferating cells; upregulation of the gene expression of cyclin dependent kinase 4 and cyclin D1, components of a holoenzyme that plays a principal role in mediating Rb phosphorylation; and marked inhibition of the gene expression of myogenin, a member of the MyoD family of skeletal muscle-specific transcription factors that is essential in muscle differentiation. We also find that IGF-induced inhibition of differentiation occurs through a process that is independent of its mitogenic effects. We demonstrate, thus, that IGFs regulate Rb phosphorylation and cyclin D1 and cyclin-dependent kinase 4 gene expression; together with their biphasic effects on myogenin expression, these results suggest a mechanism by which IGFs are initially mitogenic and subsequently differentiation-promoting in skeletal muscle. PMID- 7479776 TI - Nonsense mutation in the phosphofructokinase muscle subunit gene associated with retention of intron 10 in one of the isolated transcripts in Ashkenazi Jewish patients with Tarui disease. AB - Mutations in the human phosphofructokinase muscle subunit gene (PFKM) are known to cause myopathy classified as glycogenosis type VII (Tarui disease). Previously described molecular defects include base substitutions altering encoded amino acids or resulting in abnormal splicing. We report a mutation resulting in phosphofructokinase deficiency in three patients from an Ashkenazi Jewish family. Using a reverse transcription PCR assay, PFKM subunit transcripts differing by length were detected in skeletal muscle tissue of all three affected subjects. In the longer transcript, an insertion of 252 nucleotides totally homologous to the structure of the 10th intron of the PFKM gene was found separating exon 10 from exon 11. In addition, two single base transitions were identified by direct sequencing: [exon 6; codon 95; CGA (Arg) to TGA (stop)] and [exon 7; codon 172; ACC (Thr) to ACT (Thr)] in either transcript. Single-stranded conformational polymorphism and restriction enzyme analyses confirmed the presence of these point substitutions in genomic DNA and strongly suggested homozygosity for the pathogenic allele. The nonsense mutation at codon 95 appeared solely responsible for the phenotype in these patients, further expanding genetic heterogeneity of Tarui disease. Transcripts with and without intron 10 arising from identical mutant alleles probably resulted from differential pre-mRNA processing and may represent a novel message from the PFKM gene. PMID- 7479775 TI - Genes that control a temperature-compensated ultradian clock in Caenorhabditis elegans. AB - Substantial progress has been made in understanding the genetic basis of temperature-compensated circadian clocks. Ultradian rhythms, with a period shorter than 24 h, are at least as widespread as circadian rhythms. We have initiated genetic analysis of defecation behavior, which is controlled by an ultradian clock in Caenorhabditis elegans. The defecation motor program is activated every 45 sec, and this rhythm is temperature compensated. We describe mutations in 12 genes that either shorten or lengthen the cycle period. We find that most of these mutations also disrupt temperature compensation, suggesting that this process is an integral part of the clock. These genes open the way for molecular genetic dissection of this ultradian clock. PMID- 7479774 TI - Modulation of cholecystokinin activity by albumin. AB - We found that a variety of cholecystokinin (CCK) receptor ligands bind to bovine serum albumin (BSA). This binding was rapid, fully reversible, temperature independent, of low affinity, and specific for BSA; it depended on the concentration of BSA, the chemical structure of the ligand, and the chemical composition of the incubation medium. BSA also decreased the binding of 125I labeled CCK octapeptide (125I-CCK-8) to CCK receptors on pancreatic acini and membranes but increased the potency with which CCK-8 inhibited binding of 125I CCK-8. These counterintuitive findings appeared to result from BSA altering the affinities of CCK-8 for different affinity states of the pancreatic CCK receptor. An alternate hypothesis is that BSA increased the efficacy of CCK-8 such that it bound to receptors and also caused biochemical changes in other receptors that reduced their ability to bind 125I-CCK-8. BSA enhanced the ability of CCK-8 to stimulate amylase secretion from pancreatic acini and to cause contraction of dispersed gastric smooth muscle cells. Thus, CCK can bind to BSA, and the BSA-CCK complex has substantially different activities from the free, uncomplexed hormone. PMID- 7479777 TI - Desensitization of N-methyl-D-aspartate receptors: a problem of interpretation. AB - The phenomenon of desensitization is universal, but its mechanism is still ill understood and controversial. A recently published study [Lin, F. & Stevens, C. F. (1994) J. Neurosci, 14, 2153-2160] attempted to cast light on the mechanism of desensitization of N-methyl-D-aspartate (NMDA) receptors, in particular the vexed question of whether the channel must open before it can desensitize. During the desensitizing preexposure to agonist in those experiments, more desensitization was produced when channel openings were observed than when no openings were observed. The conclusion that "desensitization occurs more rapidly from the open state" unfortunately was based on a stochastic fallacy, and we present here a theoretical treatment and illustration showing that the observed behavior is predicted by a simple mechanism in which desensitization can occur only from a shut state. PMID- 7479778 TI - Transcription factor TFIID is a direct functional target of the adenovirus E1A transcription-repression domain. AB - The 243-amino acid adenovirus E1A oncoprotein both positively and negatively modulates the expression of cellular genes involved in the regulation of cell growth. The E1A transcription repression function appears to be linked with its ability to induce cellular DNA synthesis, cell proliferation, and cell transformation, as well as to inhibit cell differentiation. The mechanism by which E1A represses the transcription of various promoters has proven enigmatic. Here we provide several lines of evidence that the "TATA-box" binding protein (TBP) component of transcription factor TFIID is a cellular target of the E1A repression function encoded within the E1A N-terminal 80 amino acids. (i) The E1A N-terminal 80 amino acids [E1A-(1-80)protein] efficiently represses basal transcription from TATA-containing core promoters in vitro. (ii) TBP reverses completely E1A repression in vitro. (iii) TBP restores transcriptional activity to E1A-(1-80) protein affinity-depleted nuclear extracts. (iv) The N-terminal repression domain of E1A interacts directly and specifically with TBP in vitro. These results may help explain how E1A represses a set of genes that lack common upstream promoter elements. PMID- 7479779 TI - In vitro integration of human immunodeficiency virus type 1 cDNA into targets containing protein-induced bends. AB - Integration of human immunodeficiency virus type 1 cDNA into a target DNA can be strongly influenced by the conformation of the target. For example, integration in vitro is sometimes favored in target DNAs containing sequence-directed bends or DNA distortions caused by bound proteins. We have analyzed the effect of DNA bending by studying integration into two well-characterized protein-DNA complexes: Escherichia coli integration host factor (IHF) protein bound to a phage IHF site, and the DNA binding domain of human lymphoid enhancer factor (LEF) bound to a LEF site. Both of these proteins have previously been reported to bend DNA by approximately 140 degrees. Binding of IHF greatly increases the efficiency of in vitro integration at hotspots within the IHF site. We analyzed a series of mutants in which the IHF site was modified at the most prominent hotspot. We found that each variant still displayed enhanced integration upon IHF binding. Evidently the local sequence is not critical for formation of an IHF hotspot. LEF binding did not create preferred sites for integration. The different effects of IHF and LEF binding can be rationalized in terms of the different proposed conformations of the two protein-DNA complexes. PMID- 7479780 TI - VIP21/caveolin is a cholesterol-binding protein. AB - VIP21/caveolin is localized to both caveolae and apical transport vesicles and presumably cycles between the cell surface and the Golgi complex. We have studied the lipid interactions of this protein by reconstituting Escherichia coli expressed VIP21/caveolin into liposomes. Surprisingly, the protein reconstituted only with cholesterol-containing lipid mixtures. We demonstrated that the protein binds at least 1 mol of cholesterol per mole of protein and that this binding promotes formation of protein oligomers. These findings suggest that VIP21/caveolin, through its cholesterol-binding properties, serves a specific function in microdomain formation during membrane trafficking. PMID- 7479781 TI - Identification of a Drosophila muscle development gene with structural homology to mammalian early growth response transcription factors. AB - In Drosophila, stripe (sr) gene function is required for normal muscle development. Some mutations disrupt embryonic muscle development and are lethal. Other mutations cause total loss of only a single muscle in the adult. Molecular analysis shows that sr encodes a predicted protein containing a zinc finger motif. This motif is homologous to the DNA binding domains encoded by members of the early growth response (egr) gene family. In mammals, expression of egr genes is induced by intercellular signals, and there is evidence for their role in many developmental events. The identification of sr as an egr gene and its pattern of expression suggest that it functions in muscle development via intercellular communication. PMID- 7479782 TI - Chloroplast DNA variation and the recent radiation of the giant senecios (Asteraceae) on the tall mountains of eastern Africa. AB - Chloroplast DNA restriction-site variation was surveyed among 40 accessions representing all 11 species of giant senecios (Dendrosenecio, Asteraceae) at all but one known location, plus three outgroup species. Remarkably little variation (only 9 variable sites out of roughly 1000 sites examined) was found among the 40 giant senecio accessions, yet as a group they differ significantly (at 18 sites) from Cineraria deltoidea, the closest known relative. This pattern indicates that the giant senecios underwent a recent dramatic radiation in eastern Africa and evolved from a relatively isolated lineage within the Senecioneae. Biogeographic interpretation of the molecular phylogeny suggests that the giant senecios originated high on Mt. Kilimanjaro, with subsequent dispersion to the Aberdares, Mt. Kenya, and the Cherangani Hills, followed by dispersion westward to the Ruwenzori Mountains, and then south to the Virunga Mountains, Mt. Kahuzi, and Mt. Muhi, but with dispersion back to Mt. Elgon. Geographic radiation was an important antecedent to the diversification in eastern Africa, which primarily involved repeated altitudinal radiation, both up and down the mountains, leading to morphological parallelism in both directions. In general, the plants on a given mountain are more closely related to each other than they are to plants on other mountains, and plants on nearby mountains are more closely related to each other than they are to plants on more distant mountains. The individual steps of the geographic radiation have occurred at various altitudes, some clearly the result of intermountain dispersal. The molecular evidence suggests that two species are extant ancestors to other species on the same or nearby mountains. PMID- 7479783 TI - Identification of an additional gene required for eukaryotic nonsense mRNA turnover. AB - Loss of function of any one of three UPF genes prevents the accelerated decay of nonsense mRNAs in Saccharomyces cerevisiae. We report the identification and DNA sequence of UPF3, which is present in one nonessential copy on chromosome VII. Upf3 contains three putative nuclear localization signal sequences, suggesting that it may be located in a different compartment than the cytoplasmic Upf1 protein. Epitope-tagged Upf3 (FLAG-Upf3) does not cofractionate with polyribosomes or 80S ribosomal particles. Double disruptions of UPF1 and UPF3 affect nonsense mRNA decay in a manner indistinguishable from single disruptions. These results suggest that the Upf proteins perform related functions in a common pathway. PMID- 7479784 TI - The cell wall components peptidoglycan and lipoteichoic acid from Staphylococcus aureus act in synergy to cause shock and multiple organ failure. AB - Although the incidence of Gram-positive sepsis has risen strongly, it is unclear how Gram-positive organisms (without endotoxin) initiate septic shock. We investigated whether two cell wall components from Staphylococcus aureus, peptidoglycan (PepG) and lipoteichoic acid (LTA), can induce the inflammatory response and multiple organ dysfunction syndrome (MODS) associated with septic shock caused by Gram-positive organisms. In cultured macrophages, LTA (10 micrograms/ml), but not PepG (100 micrograms/ml), induces the release of nitric oxide measured as nitrite. PepG, however, caused a 4-fold increase in the production of nitrite elicited by LTA. Furthermore, PepG antibodies inhibited the release of nitrite elicited by killed S. aureus. Administration of both PepG (10 mg/kg; i.v.) and LTA (3 mg/kg; i.v.) in anesthetized rats resulted in the release of tumor necrosis factor alpha and interferon gamma and MODS, as indicated by a decrease in arterial oxygen pressure (lung) and an increase in plasma concentrations of bilirubin and alanine aminotransferase (liver), creatinine and urea (kidney), lipase (pancreas), and creatine kinase (heart or skeletal muscle). There was also the expression of inducible nitric oxide synthase in these organs, circulatory failure, and 50% mortality. These effects were not observed after administration of PepG or LTA alone. Even a high dose of LTA (10 mg/kg) causes only circulatory failure but no MODS. Thus, our results demonstrate that the two bacterial wall components, PepG and LTA, work together to cause systemic inflammation and multiple systems failure associated with Gram-positive organisms. PMID- 7479786 TI - Functional implications for the microtubule-associated protein tau: localization in oligodendrocytes. AB - We present evidence that the microtubule-associated protein tau is present in oligodendrocytes (OLGs), the central nervous system cells that make myelin. By showing that tau is distributed in a pattern similar to that of myelin basic protein, our results suggest a possible involvement of tau in some aspect of myelination. Tau protein has been identified in OLGs in situ and in vitro. In interfascicular OLGs, tau localization, revealed by monoclonal antibody Tau-5, was confined to the cell somata. However, in cultured ovine OLGs with an exuberant network of processes, tau was detected in cell somata, cellular processes, and membrane expansions at the tips of these processes. Moreover, in such cultures, tau appeared localized adjacent to or coincident with myelin basic protein in membrane expansions along and at the ends of the cellular processes. The presence of tau mRNA was documented using fluorescence in situ hybridization. The distribution of the tau mRNA was similar to that of the tau protein. Western blot analysis of cultured OLGs showed the presence of many tau isoforms. Together, these results demonstrate that tau is a genuine oligodendrocyte protein and pave the way for determining its functional role in these cells. PMID- 7479785 TI - Interleukin 1 beta suppresses transforming growth factor-induced inorganic pyrophosphate (PPi) production and expression of the PPi-generating enzyme PC-1 in human chondrocytes. AB - Articular cartilage chondrocytes have the unique ability to elaborate large amounts of extracellular pyrophosphate (PPi), and transforming growth factor beta (TGF beta) appears singular among cartilage regulatory factors in stimulating PPi production. TGF beta caused a time and dose-dependent increase in intracellular and extracellular PPi in human articular chondrocyte cultures. TGF beta and interleukin 1 beta (IL-1 beta) antagonistically regulate certain chondrocyte functions. IL-1 beta profoundly inhibited basal and TGF beta-induced PPi elaboration. To address mechanisms involved with the regulation of PPi synthesis by IL-1 beta and TGF beta, we analyzed the activity of the PPi-generating enzyme NTP pyrophosphohydrolase (NTPPPH) and the PPi-hydrolyzing enzyme alkaline phosphatase. Human chondrocyte NTPPPH activity was largely attributable to plasma cell membrane glycoprotein 1, PC-1. Furthermore, TGF beta induced comparable increases in the activity of extracellular PPi, intracellular PPi, and cellular NTPPPH and in the levels of PC-1 protein and mRNA in chondrocytes as well as a decrease in alkaline phosphatase. All of these TGF beta-induced responses were completely blocked by IL-1 beta. Thus, IL-1 beta may be an important regulator of mineralization in chondrocytes by inhibiting TGF beta-induced PPi production and PC-1 expression. PMID- 7479787 TI - Genetic rearrangements in the rfb regions of Vibrio cholerae O1 and O139. AB - The recent emergence of a pathogenic new non-O1 serotype (O139) of Vibrio cholerae has led to numerous studies in an attempt to identify the origins of this new strain. Our studies indicate that O139 strains have clear differences in the surface polysaccharides when compared with O1 strains: the lipopolysaccharide can be described as semi-rough. Southern hybridization with the O1 rfb region demonstrates that O139 strains no longer contain any of the rfb genes required for the synthesis of the O1 O-antigen or its modification and also lack at least 6 kb of additional contiguous DNA. However, O139 strains have retained rfaD and have a single open reading frame closely related to three small open reading frames of the O1 rfb region. This region is closely related to the H-repeat of Escherichia coli and to the transposases of a number of insertion sequence elements and has all the features of an insertion sequence element that has been designated VcIS1. Transposon insertion mutants defective in O139 O-antigen (and capsule) biosynthesis map to the same fragment as VcIS1. Preliminary sequence data of complementing clones indicate that this DNA encodes a galactosyl transferase and other enzymes for the utilization of galactose in polysaccharide biosynthesis. We propose a mechanism by which both the Ogawa serotype of O1 strains and the O139 serotype strains may have evolved. PMID- 7479789 TI - Electrostatic force microscope for probing surface charges in aqueous solutions. AB - A scanning force microscope was converted to an electrostatic force microscope by charging the usually neutral cantilever with phospholipids. The electrostatic force microscope was used to study surface electrostatic charges of samples in aqueous solutions. Lysozymes, DEAE-Sephadex beads, 3-propyltriethoxysilane treated glass and mica were imaged in water or phosphate buffer with electrostatic force microscopy. The adhesion force measured when a charged probe and oppositely charged specimen interacted was up to 500 times greater than when a bare probe was used. This dramatic increase in measured adhesion force can be attributed to the energy required to break the salt bridges formed between the charged probe and the specimen. The use of phospholipids to functionalize the cantilever tip allows the incorporation of other biomolecules and ligands that can be used as biologically specific tips (e.g., receptors, drugs) for the study of intermolecular interactions. PMID- 7479790 TI - Cyclic polyamides for recognition in the minor groove of DNA. AB - Small molecules that specifically bind with high affinity to any designated DNA sequence in the human genome would be useful tools in molecular biology and potentially in human medicine. Simple rules have been developed to rationally alter the sequence specificity of minor groove-binding polyamides containing N methylimidazole and N-methylpyrrole amino acids. Crescent-shaped polyamides bind as antiparallel dimers with each polyamide making specific contacts with each strand on the floor of the minor groove. Cyclic polyamides have now been synthesized that bind designated DNA sequences at subnanomolar concentrations. PMID- 7479788 TI - ndhF sequence evolution and the major clades in the sunflower family. AB - An extensive sequence comparison of the chloroplast ndhF gene from all major clades of the largest flowering plant family (Asteraceae) shows that this gene provides approximately 3 times more phylogenetic information than rbcL. This is because it is substantially longer and evolves twice as fast. The 5' region (1380 bp) of ndhF is very different from the 3' region (855 bp) and is similar to rbcL in both the rate and the pattern of sequence change. The 3' region is more A+T rich, has higher levels of nonsynonymous base substitution, and shows greater transversion bias at all codon positions. These differences probably reflect different functional constraints on the 5' and 3' regions of ndhF. The two patterns of base substitutions of ndhF are particularly advantageous for phylogenetic reconstruction because the conserved and variable segments can be used for older and recent groups, respectively. Phylogenetic analyses of 94 ndhF sequences provided much better resolution of relationships than previous molecular and morphological phylogenies of the Asteraceae. The ndhF tree identified five major clades: (i) the Calyceraceae is the sister family of Asteraceae; (ii) the Barnadesioideae is monophyletic and is the sister group to the rest of the family; (iii) the Cichorioideae and its two basal tribes Mutisieae and Cardueae are paraphyletic; (iv) four tribes of Cichorioideae (Lactuceae, Arctoteae, Liabeae, and Vernonieae) form a monophyletic group, and these are the sister clade of the Asteroideae; and (v) the Asteroideae is monophyletic and includes three major clades. PMID- 7479791 TI - RecA.oligonucleotide filaments bind in the minor groove of double-stranded DNA. AB - Escherichia coli RecA protein, in the presence of ATP or its analog adenosine 5' [gamma-thio]triphosphate, polymerizes on single-stranded DNA to form nucleoprotein filaments that can then bind to homologous sequences on duplex DNA. The three-stranded joint molecule formed as a result of this binding event is a key intermediate in general recombination. We have used affinity cleavage to examine this three-stranded joint by incorporating a single thymidine-EDTA.Fe (T*) into the oligonucleotide part of the filament. Our analysis of the cleavage patterns from the joint molecule reveals that the nucleoprotein filament binds in the minor groove of an extended Watson-Crick duplex. PMID- 7479792 TI - Similar antigenic surfaces, rather than sequence homology, dictate T-cell epitope molecular mimicry. AB - Molecular mimicry, normally defined by the level of primary-sequence similarities between self and foreign antigens, has been considered a key element in the pathogenesis of autoimmunity. Here we describe an example of molecular mimicry between two overlapping peptides within a single self-antigen, both of which are recognized by the same human self-reactive T-cell clone. Two intervening peptides did not stimulate the T-cell clone, even though they share nine amino acids with the stimulatory peptides. Molecular modeling of major histocompatibility complex class II-peptide complexes suggests that both of the recognized peptides generate similar antigenic surfaces, although these are composed of different sets of amino acids. The molecular modeling of a peptide shifted one residue from the stimulatory peptide, which was recognized in the context of the same HLA molecule by another T-cell clone, generated a completely different antigenic surface. Functional studies using truncated peptides confirmed that the anchor residues of the two "mimicking" epitopes in the HLA groove differ. Our results show, for two natural epitopes, how molecular mimicry can occur and suggest that studies of potential antigenic surfaces, rather than sequence similarity, are necessary for analyzing suspected peptide mimicry. PMID- 7479793 TI - Y chromosome short arm-Sxr recombination in XSxr/Y males causes deletion of Rbm and XY female sex reversal. AB - We earlier described three lines of sex-reversed XY female mice deleted for sequences believed close to the testes-determining gene (Sry) on the Y chromosome short arm (Yp). The original sex-reversed females appeared among the offspring of XY males that carried the Yp duplication Sxr on their X chromosome. Earlier cytogenetic observations had suggested that the deletions resulted from asymmetrical meiotic recombination between the Y and the homologous Sxr region, but no direct evidence for this hypothesis was available. We have now analyzed the offspring of XSxr/Y males carrying an evolutionarily divergent Mus musculus domesticus Y chromosome, which permits detection and characterization of such recombination events. This analysis has enabled the derivation of a recombination map of Yp and Sxr, also demonstrating the orientation of Yp with respect to the Y centromere. The mapping data have established that Rbm, the murine homologue of a gene family cloned from the human Y chromosome, lies between Sry and the centromere. Analysis of two additional XY female lines shows that asymmetrical Yp Sxr recombination leading to XY female sex reversal results in deletion of Rbm sequences. The deletions bring Sry closer to Y centromere, consistent with the hypothesis that position-effect inactivation of Sry is the basis for the sex reversal. PMID- 7479794 TI - Muscle-specific expression of Drosophila hsp70 in response to aging and oxidative stress. AB - Induction of Drosophila hsp70 protein was detected during aging in flight muscle and leg muscle in the absence of heat shock, using an hsp70-specific monoclonal antibody, and in transgenic flies containing hsp70-beta-galactosidase fusion protein reporter constructs. While hsp70 and reporter proteins were induced during aging, hsp70 message levels were not, indicating that aging-specific induction is primarily posttranscriptional. In contrast, hsp22 and hsp23 were found to be induced during aging at the RNA level and with a broader tissue distribution. The same muscle-specific hsp70 reporter expression pattern was observed in young flies mutant for catalase (H2O2:H2O2 oxidoreductase, EC 1.11.1.6). In catalase (cat) hypomorphic lines where flies survived to older ages, the time course of hsp70 reporter expression during aging was accelerated, and the initial and ultimate levels of expression were increased. The hsp70 reporter was also induced in young flies mutant for copper/zinc superoxide dismutase (superoxide:superoxide oxidoreductase, EC 1.15.1.1). Taken together, the results suggest that aging-specific hsp70 expression may be a result of oxidative damage. PMID- 7479796 TI - Mutations in the MSH3 gene preferentially lead to deletions within tracts of simple repetitive DNA in Saccharomyces cerevisiae. AB - Eukaryotic genomes contain tracts of DNA in which a single base or a small number of bases are repeated (microsatellites). Mutations in the yeast DNA mismatch repair genes MSH2, PMS1, and MLH1 increase the frequency of mutations for normal DNA sequences and destabilize microsatellites. Mutations of human homologs of MSH2, PMS1, and MLH1 also cause microsatellite instability and result in certain types of cancer. We find that a mutation in the yeast gene MSH3 that does not substantially affect the rate of spontaneous mutations at several loci increases microsatellite instability about 40-fold, preferentially causing deletions. We suggest that MSH3 has different substrate specificities than the other mismatch repair proteins and that the human MSH3 homolog (MRP1) may be mutated in some tumors with microsatellite instability. PMID- 7479795 TI - Benzoic acid 2-hydroxylase, a soluble oxygenase from tobacco, catalyzes salicylic acid biosynthesis. AB - Benzoic acid 2-hydroxylase (BA2H) catalyzes the biosynthesis of salicylic acid from benzoic acid. The enzyme has been partially purified and characterized as a soluble protein of 160 kDa. High-efficiency in vivo labeling of salicylic acid with 18O2 suggested that BA2H is an oxygenase that specifically hydroxylates the ortho position of benzoic acid. The enzyme was strongly induced by either tobacco mosaic virus inoculation or benzoic acid infiltration of tobacco leaves and it was inhibited by CO and other inhibitors of cytochrome P450 hydroxylases. The BA2H activity was immunodepleted by antibodies raised against SU2, a soluble cytochrome P450 from Streptomyces griseolus. The anti-SU2 antibodies immunoprecipitated a radiolabeled polypeptide of around 160 kDa from the soluble protein extracts of L-[35S]-methionine-fed tobacco leaves. Purified BA2H showed CO-difference spectra with a maximum at 457 nm. These data suggest that BA2H belongs to a novel class of soluble, high molecular weight cytochrome P450 enzymes. PMID- 7479797 TI - Relating aromatic hydrocarbon-induced DNA adducts and c-H-ras mutations in mouse skin papillomas: the role of apurinic sites. AB - Mouse skin tumors contain activated c-H-ras oncogenes, often caused by point mutations at codons 12 and 13 in exon 1 and codons 59 and 61 in exon 2. Mutagenesis by the noncoding apurinic sites can produce G-->T and A-->T transversions by DNA misreplication with more frequent insertion of deoxyadenosine opposite the apurinic site. Papillomas were induced in mouse skin by several aromatic hydrocarbons, and mutations in the c-H-ras gene were determined to elucidate the relationship among DNA adducts, apurinic sites, and ras oncogene mutations. Dibenzo[a,l]pyrene (DB[a,l]P), DB[a,l]P-11,12 dihydrodiol, anti-DB[a,l]P-11,12-diol-13,14-epoxide, DB[a,l]P-8,9-dihydrodiol, 7,12-dimethylbenz[a]anthracene (DMBA), and 1,2,3,4-tetrahydro-DMBA consistently induced a CAA-->CTA mutation in codon 61 of the c-H-ras oncogene. Benzo[a]pyrene induced a GGC-->GTC mutation in codon 13 in 54% of tumors and a CAA-->CTA mutation in codon 61 in 15%. The pattern of mutations induced by each hydrocarbon correlated with its profile of DNA adducts. For example, both DB[a,l]P and DMBA primarily form DNA adducts at the N-3 and/or N-7 of deoxyadenosine that are lost from the DNA by depurination, generating apurinic sites. Thus, these results support the hypothesis that misreplication of unrepaired apurinic sites generated by loss of hydrocarbon-DNA adducts is responsible for transforming mutations leading to papillomas in mouse skin. PMID- 7479798 TI - Cloning and characterization of a cellular apoptosis susceptibility gene, the human homologue to the yeast chromosome segregation gene CSE1. AB - We recently isolated human cDNA fragments that render MCF-7 breast cancer cells resistant to cell death caused by Pseudomonas exotoxin, Pseudomonas exotoxin derived immunotoxins, diphtheria toxin, and tumor necrosis factor. We report here that one of these fragments is an antisense fragment of a gene homologous to the essential yeast chromosome segregation gene CSE1. Cloning and analysis of the full-length cDNA of the human CSE1 homologue, which we name CAS for cellular apoptosis susceptibility gene, reveals a protein coding region with similar length (971 amino acids for CAS, 960 amino acids for CSE1) and 59% overall protein homology to the yeast CSE1 protein. The conservation of this gene indicates it has an important function in human cells consistent with the essential role of CSE1 in yeast. CAS is highly expressed in human tumor cell lines and in human testis and fetal liver, tissues that contain actively dividing cells. Furthermore, CAS expression increases when resting human fibroblasts are induced to proliferate and decreases when they are growth-arrested. Thus, CAS appears to play an important role in both toxin and tumor necrosis factor mediated cell death, as well as in cell proliferation. PMID- 7479799 TI - Transfer RNA editing in land snail mitochondria. AB - Some mitochondrial tRNA genes of land snails show mismatches in the acceptor stems predicted from their gene sequences. The majority of these mismatches fall in regions where the tRNA genes overlap with adjacent downstream genes. We have synthesized cDNA from four circularized tRNAs and determined the sequences of the 5' and 3' parts of their acceptor stems. Three of the four tRNAs differ from their corresponding genes at a total of 13 positions, which all fall in the 3' part of the acceptor stems as well as the discriminator bases. The editing events detected involve changes from cytidine, thymidine, and guanosine to adenosine residues, which generally restore base-pairing in the stems. However, in one case an A-A mismatch is created from an A-C mismatch. It is suggested that this form of RNA editing may involve polyadenylylation of the maturing tRNAs as an intermediate. PMID- 7479801 TI - Voice communication between humans and machines--an introduction. PMID- 7479802 TI - Scientific bases of human-machine communication by voice. AB - The scientific bases for human-machine communication by voice are in the fields of psychology, linguistics, acoustics, signal processing, computer science, and integrated circuit technology. The purpose of this paper is to highlight the basic scientific and technological issues in human-machine communication by voice and to point out areas of future research opportunity. The discussion is organized around the following major issues in implementing human-machine voice communication systems: (i) hardware/software implementation of the system, (ii) speech synthesis for voice output, (iii) speech recognition and understanding for voice input, and (iv) usability factors related to how humans interact with machines. PMID- 7479800 TI - Lack of functional retinoblastoma protein mediates increased resistance to antimetabolites in human sarcoma cell lines. AB - Growth inhibition assays indicated that the IC50 values for methotrexate (MTX) and 5-fluorodeoxyuridine (FdUrd) in HS-18, a liposarcoma cell line lacking retinoblastoma protein (pRB), and SaOS-2, an osteosarcoma cell line with a truncated and nonfunctional pRB, were 10- to 12-fold and 4- to 11-fold higher, respectively, than for the HT-1080 (fibrosarcoma) cell line, which has wild-type pRB. These Rb-/- cell lines exhibited a 2- to 4-fold increase in both dihydrofolate reductase (DHFR) and thymidylate synthase (TS) enzyme activities as well as a 3- to 4-fold increase in mRNA levels for these enzymes compared to the HT-1080 (Rb+/+) cells. This increase in expression was not due to amplification of the DHFR and TS genes. Growth inhibition by MTX and FdUrd was increased and DHFR and TS activities and expression were correspondingly decreased in Rb transfectants of SaOS-2 cells. In contrast, there was no significant difference in growth inhibition among these cell lines for the nonantimetabolites VP-16, cisplatin, and doxorubicin. A gel mobility-shift assay showed that parental SaOS 2 cells had increased levels of free E2F compared to the Rb-reconstituted SaOS-2 cells. These results indicate that pRB defective cells may have decreased sensitivity to growth inhibition by target enzymes encoded by genes whose transcription is enhanced by E2F proteins and suggest mechanisms of interaction between cytotoxic agents and genes involved in cell cycle progression. PMID- 7479803 TI - The role of voice input for human-machine communication. AB - Optimism is growing that the near future will witness rapid growth in human computer interaction using voice. System prototypes have recently been built that demonstrate speaker-independent real-time speech recognition, and understanding of naturally spoken utterances with vocabularies of 1000 to 2000 words, and larger. Already, computer manufacturers are building speech recognition subsystems into their new product lines. However, before this technology can be broadly useful, a substantial knowledge base is needed about human spoken language and performance during computer-based spoken interaction. This paper reviews application areas in which spoken interaction can play a significant role, assesses potential benefits of spoken interaction with machines, and compares voice with other modalities of human-computer interaction. It also discusses information that will be needed to build a firm empirical foundation for the design of future spoken and multimodal interfaces. Finally, it argues for a more systematic and scientific approach to investigating spoken input and performance with future language technology. PMID- 7479804 TI - Computer speech synthesis: its status and prospects. AB - Computer speech synthesis has reached a high level of performance, with increasingly sophisticated models of linguistic structure, low error rates in text analysis, and high intelligibility in synthesis from phonemic input. Mass market applications are beginning to appear. However, the results are still not good enough for the ubiquitous application that such technology will eventually have. A number of alternative directions of current research aim at the ultimate goal of fully natural synthetic speech. One especially promising trend is the systematic optimization of large synthesis systems with respect to formal criteria of evaluation. Speech recognition has progressed rapidly in the past decade through such approaches, and it seems likely that their application in synthesis will produce similar improvements. PMID- 7479805 TI - Models of speech synthesis. AB - The term "speech synthesis" has been used for diverse technical approaches. In this paper, some of the approaches used to generate synthetic speech in a text-to speech system are reviewed, and some of the basic motivations for choosing one method over another are discussed. It is important to keep in mind, however, that speech synthesis models are needed not just for speech generation but to help us understand how speech is created, or even how articulation can explain language structure. General issues such as the synthesis of different voices, accents, and multiple languages are discussed as special challenges facing the speech synthesis community. PMID- 7479806 TI - Research in speech communication. AB - Advances in digital speech processing are now supporting application and deployment of a variety of speech technologies for human/machine communication. In fact, new businesses are rapidly forming about these technologies. But these capabilities are of little use unless society can afford them. Happily, explosive advances in microelectronics over the past two decades have assured affordable access to this sophistication as well as to the underlying computing technology. The research challenges in speech processing remain in the traditionally identified areas of recognition, synthesis, and coding. These three areas have typically been addressed individually, often with significant isolation among the efforts. But they are all facets of the same fundamental issue--how to represent and quantify the information in the speech signal. This implies deeper understanding of the physics of speech production, the constraints that the conventions of language impose, and the mechanism for information processing in the auditory system. In ongoing research, therefore, we seek more accurate models of speech generation, better computational formulations of language, and realistic perceptual guides for speech processing--along with ways to coalesce the fundamental issues of recognition, synthesis, and coding. Successful solution will yield the long-sought dictation machine, high-quality synthesis from text, and the ultimate in low bit-rate transmission of speech. It will also open the door to language-translating telephony, where the synthetic foreign translation can be in the voice of the originating talker. PMID- 7479807 TI - Linguistic aspects of speech synthesis. AB - The conversion of text to speech is seen as an analysis of the input text to obtain a common underlying linguistic description, followed by a synthesis of the output speech waveform from this fundamental specification. Hence, the comprehensive linguistic structure serving as the substrate for an utterance must be discovered by analysis from the text. The pronunciation of individual words in unrestricted text is determined by morphological analysis or letter-to-sound conversion, followed by specification of the word-level stress contour. In addition, many text character strings, such as titles, numbers, and acronyms, are abbreviations for normal words, which must be derived. To further refine these pronunciations and to discover the prosodic structure of the utterance, word part of speech must be computed, followed by a phrase-level parsing. From this structure the prosodic structure of the utterance can be determined, which is needed in order to specify the durational framework and fundamental frequency contour of the utterance. In discourse contexts, several factors such as the specification of new and old information, contrast, and pronominal reference can be used to further modify the prosodic specification. When the prosodic correlates have been computed and the segmental sequence is assembled, a complete input suitable for speech synthesis has been determined. Lastly, multilingual systems utilizing rule frameworks are mentioned, and future directions are characterized. PMID- 7479808 TI - Speech recognition technology: a critique. AB - This paper introduces the session on advanced speech recognition technology. The two papers comprising this session argue that current technology yields a performance that is only an order of magnitude in error rate away from human performance and that incremental improvements will bring us to that desired level. I argue that, to the contrary, present performance is far removed from human performance and a revolution in our thinking is required to achieve the goal. It is further asserted that to bring about the revolution more effort should be expended on basic research and less on trying to prematurely commercialize a deficient technology. PMID- 7479809 TI - State of the art in continuous speech recognition. AB - In the past decade, tremendous advances in the state of the art of automatic speech recognition by machine have taken place. A reduction in the word error rate by more than a factor of 5 and an increase in recognition speeds by several orders of magnitude (brought about by a combination of faster recognition search algorithms and more powerful computers), have combined to make high-accuracy, speaker-independent, continuous speech recognition for large vocabularies possible in real time, on off-the-shelf workstations, without the aid of special hardware. These advances promise to make speech recognition technology readily available to the general public. This paper focuses on the speech recognition advances made through better speech modeling techniques, chiefly through more accurate mathematical modeling of speech sounds. PMID- 7479810 TI - Training and search methods for speech recognition. AB - Speech recognition involves three processes: extraction of acoustic indices from the speech signal, estimation of the probability that the observed index string was caused by a hypothesized utterance segment, and determination of the recognized utterance via a search among hypothesized alternatives. This paper is not concerned with the first process. Estimation of the probability of an index string involves a model of index production by any given utterance segment (e.g., a word). Hidden Markov models (HMMs) are used for this purpose [Makhoul, J. & Schwartz, R. (1995) Proc. Natl. Acad. Sci. USA 92, 9956-9963]. Their parameters are state transition probabilities and output probability distributions associated with the transitions. The Baum algorithm that obtains the values of these parameters from speech data via their successive reestimation will be described in this paper. The recognizer wishes to find the most probable utterance that could have caused the observed acoustic index string. That probability is the product of two factors: the probability that the utterance will produce the string and the probability that the speaker will wish to produce the utterance (the language model probability). Even if the vocabulary size is moderate, it is impossible to search for the utterance exhaustively. One practical algorithm is described [Viterbi, A. J. (1967) IEEE Trans. Inf. Theory IT-13, 260-267] that, given the index string, has a high likelihood of finding the most probable utterance. PMID- 7479811 TI - The roles of language processing in a spoken language interface. AB - This paper provides an overview of the colloquium's discussion session on natural language understanding, which followed presentations by M. Bates [Bates, M. (1995) Proc. Natl. Acad. Sci. USA 92, 9977-9982] and R. C. Moore [Moore, R. C. (1995) Proc. Natl. Acad. Sci. USA 92, 9983-9988]. The paper reviews the dual role of language processing in providing understanding of the spoken input and an additional source of constraint in the recognition process. To date, language processing has successfully provided understanding but has provided only limited (and computationally expensive) constraint. As a result, most current systems use a loosely coupled, unidirectional interface, such as N-best or a word network, with natural language constraints as a postprocess, to filter or resort the recognizer output. However, the level of discourse context provides significant constraint on what people can talk about and how things can be referred to; when the system becomes an active participant, it can influence this order. But sources of discourse constraint have not been extensively explored, in part because these effects can only be seen by studying systems in the context of their use in interactive problem solving. This paper argues that we need to study interactive systems to understand what kinds of applications are appropriate for the current state of technology and how the technology can move from the laboratory toward real applications. PMID- 7479812 TI - Models of natural language understanding. AB - This paper surveys some of the fundamental problems in natural language (NL) understanding (syntax, semantics, pragmatics, and discourse) and the current approaches to solving them. Some recent developments in NL processing include increased emphasis on corpus-based rather than example- or intuition-based work, attempts to measure the coverage and effectiveness of NL systems, dealing with discourse and dialogue phenomena, and attempts to use both analytic and stochastic knowledge. Critical areas for the future include grammars that are appropriate to processing large amounts of real language; automatic (or at least semi-automatic) methods for deriving models of syntax, semantics, and pragmatics; self-adapting systems; and integration with speech processing. Of particular importance are techniques that can be tuned to such requirements as full versus partial understanding and spoken language versus text. Portability (the ease with which one can configure an NL system for a particular application) is one of the largest barriers to application of this technology. PMID- 7479813 TI - Integration of speech with natural language understanding. AB - The integration of speech recognition with natural language understanding raises issues of how to adapt natural language processing to the characteristics of spoken language; how to cope with errorful recognition output, including the use of natural language information to reduce recognition errors; and how to use information from the speech signal, beyond just the sequence of words, as an aid to understanding. This paper reviews current research addressing these questions in the Spoken Language Program sponsored by the Advanced Research Projects Agency (ARPA). I begin by reviewing some of the ways that spontaneous spoken language differs from standard written language and discuss methods of coping with the difficulties of spontaneous speech. I then look at how systems cope with errors in speech recognition and at attempts to use natural language information to reduce recognition errors. Finally, I discuss how prosodic information in the speech signal might be used to improve understanding. PMID- 7479814 TI - A perspective on early commercial applications of voice-processing technology for telecommunications and aids for the handicapped. AB - The Colloquium on Human-Machine Communication by Voice highlighted the global technical community's focus on the problems and promise of voice-processing technology, particularly, speech recognition and speech synthesis. Clearly, there are many areas in both the research and development of these technologies that can be advanced significantly. However, it is also true that there are many applications of these technologies that are capable of commercialization now. Early successful commercialization of new technology is vital to ensure continuing interest in its development. This paper addresses efforts to commercialize speech technologies in two markets: telecommunications and aids for the handicapped. PMID- 7479815 TI - Voice-processing technologies--their application in telecommunications. AB - As the telecommunications industry evolves over the next decade to provide the products and services that people will desire, several key technologies will become commonplace. Two of these, automatic speech recognition and text-to-speech synthesis, will provide users with more freedom on when, where, and how they access information. While these technologies are currently in their infancy, their capabilities are rapidly increasing and their deployment in today's telephone network is expanding. The economic impact of just one application, the automation of operator services, is well over $100 million per year. Yet there still are many technical challenges that must be resolved before these technologies can be deployed ubiquitously in products and services throughout the worldwide telephone network. These challenges include: (i) High level of accuracy. The technology must be perceived by the user as highly accurate, robust, and reliable. (ii) Easy to use. Speech is only one of several possible input/output modalities for conveying information between a human and a machine, much like a computer terminal or Touch-Tone pad on a telephone. It is not the final product. Therefore, speech technologies must be hidden from the user. That is, the burden of using the technology must be on the technology itself. (iii) Quick prototyping and development of new products and services. The technology must support the creation of new products and services based on speech in an efficient and timely fashion. In this paper I present a vision of the voice processing industry with a focus on the areas with the broadest base of user penetration: speech recognition, text-to-speech synthesis, natural language processing, and speaker recognition technologies. The current and future applications of these technologies in the telecommunications industry will be examined in terms of their strengths, limitations, and the degree to which user needs have been or have yet to be met. Although noteworthy gains have been made in areas with potentially small user bases and in the more mature speech-coding technologies, these subjects are outside the scope of this paper. PMID- 7479816 TI - Processing of speech signals for physical and sensory disabilities. AB - Assistive technology involving voice communication is used primarily by people who are deaf, hard of hearing, or who have speech and/or language disabilities. It is also used to a lesser extent by people with visual or motor disabilities. A very wide range of devices has been developed for people with hearing loss. These devices can be categorized not only by the modality of stimulation [i.e., auditory, visual, tactile, or direct electrical stimulation of the auditory nerve (auditory-neural)] but also in terms of the degree of speech processing that is used. At least four such categories can be distinguished: assistive devices (a) that are not designed specifically for speech, (b) that take the average characteristics of speech into account, (c) that process articulatory or phonetic characteristics of speech, and (d) that embody some degree of automatic speech recognition. Assistive devices for people with speech and/or language disabilities typically involve some form of speech synthesis or symbol generation for severe forms of language disability. Speech synthesis is also used in text-to speech systems for sightless persons. Other applications of assistive technology involving voice communication include voice control of wheelchairs and other devices for people with mobility disabilities. PMID- 7479817 TI - Meiosis in Drosophila: seeing is believing. AB - Recently many exciting advances have been achieved in our understanding of Drosophila meiosis due to combined cytological and genetic approaches. New techniques have permitted the characterization of chromosome position and spindle formation in female meiosis I. The proteins encoded by the nod and ncd genes, two genes known to be needed for the proper partitioning of chromosomes lacking exchange events, have been identified and found to be kinesin-like motors. The effects of mutations in these genes on the spindle and chromosomes, together with the localization of the proteins, have yielded a model for the mechanism of female meiosis I. In male meiosis I, the chromosomal regions responsible for homolog pairing have been resolved to the level of specific DNA sequences. This provides a foundation for elucidating the molecular basis of meiotic pairing. The cytological techniques available in Drosophila also have permitted inroads into the regulation of sister-chromatid segregation. The products of two genes (mei S332 and ord) essential for sister-chromatid cohesion have been identified recently. Additional advances in understanding Drosophila meiosis are the delineation of a functional centromere by using minichromosome derivatives and the identification of several regulatory genes for the meiotic cell cycle. PMID- 7479818 TI - Sex and the single cell: meiosis in yeast. AB - Recent studies of Saccharomyces cerevisiae have significantly advanced our understanding of the molecular mechanisms of meiotic chromosome behavior. Structural components of the synaptonemal complex have been identified and studies of mutants defective in synapsis have provided insight into the role of the synaptonemal complex in homolog pairing, genetic recombination, crossover interference, and meiotic chromosome segregation. There is compelling evidence that most or all meiotic recombination events initiate with double-strand breaks. Several intermediates in the double-strand break repair pathway have been characterized and mutants blocked at different steps in the pathway have been identified. With the application of genetic, molecular, cytological, and biochemical methods in a single organism, we can expect an increasingly comprehensive and unified view of the meiotic process. PMID- 7479819 TI - Suppression of retinal neovascularization in vivo by inhibition of vascular endothelial growth factor (VEGF) using soluble VEGF-receptor chimeric proteins. AB - The majority of severe visual loss in the United States results from complications associated with retinal neovascularization in patients with ischemic ocular diseases such as diabetic retinopathy, retinal vein occlusion, and retinopathy of prematurity. Intraocular expression of the angiogenic protein vascular endothelial growth factor (VEGF) is closely correlated with neovascularization in these human disorders and with ischemia-induced retinal neovascularization in mice. In this study, we evaluated whether in vivo inhibition of VEGF action could suppress retinal neovascularization in a murine model of ischemic retinopathy. VEGF-neutralizing chimeric proteins were constructed by joining the extracellular domain of either human (Flt) or mouse (Flk) high-affinity VEGF receptors with IgG. Control chimeric proteins that did not bind VEGF were also used. VEGF-receptor chimeric proteins eliminated in vitro retinal endothelial cell growth stimulation by either VEGF (P < 0.006) or hypoxic conditioned medium (P < 0.005) without affecting growth under nonstimulated conditions. Control proteins had no effect. To assess in vivo response, animals with bilateral retinal ischemia received intravitreal injections of VEGF antagonist in one eye and control protein in the contralateral eye. Retinal neovascularization was quantitated histologically by a masked protocol. Retinal neovascularization in the eye injected with human Flt or murine Flk chimeric protein was reduced in 100% (25/25; P < 0.0001) and 95% (21/22; P < 0.0001) 0.0001) of animals, respectively, compared to the control treated eye. This response was evident after only a single intravitreal injection and was dose dependent with suppression of neovascularization noted after total delivery of 200 ng of protein (P < 0.002). Reduction of histologically evident neovascular nuclei per 6-microns section averaged 47% +/- 4% (P < 0.001) and 37% +/- 2% (P < 0.001) for Flt and Flk chimeric proteins with maximal inhibitory effects of 77% and 66%, respectively. No retinal toxicity was observed by light microscopy. These data demonstrate VEGF's causal role in retinal angiogenesis and prove the potential of VEGF inhibition as a specific therapy for ischemic retinal disease. PMID- 7479820 TI - Proteolytic maturation of protein C upon engineering the mouse mammary gland to express furin. AB - Endoproteolytic processing of the human protein C (HPC) precursor to its mature form involves cleavage of the propeptide after amino acids Lys-2-Arg-1 and removal of a Lys156-Arg157 dipeptide connecting the light and heavy chains. This processing was inefficient in the mammary gland of transgenic mice and pigs. We hypothesized that the protein processing capacity of specific animal organs may be improved by the coexpression of selected processing enzymes. We tested this by targeting expression of the human proprotein processing enzyme, named paired basic amino acid cleaving enzyme (PACE)/furin, or an enzymatically inactive mutant, PACEM, to the mouse mammary gland. In contrast to mice expressing HPC alone, or to HPC/PACEM bigenic mice, coexpression of PACE with HPC resulted in efficient conversion of the precursor to mature protein, with cleavage at the appropriate sites. These results suggest the involvement of PACE in the processing of HPC in vivo and represent an example of the engineering of animal organs into bioreactors with enhanced protein processing capacity. PMID- 7479821 TI - Molecular cloning and characterization of a cellular phosphoprotein that interacts with a conserved C-terminal domain of adenovirus E1A involved in negative modulation of oncogenic transformation. AB - The adenovirus type 2/5 E1A proteins transform primary baby rat kidney (BRK) cells in cooperation with the activated Ras (T24 ras) oncoprotein. The N-terminal half of E1A (exon 1) is essential for this transformation activity. While the C terminal half of E1A (exon 2) is dispensable, a region located between residues 225 and 238 of the 243R E1A protein negatively modulates in vitro T24 ras cooperative transformation as well as the tumorigenic potential of E1A/T24 ras transformed cells. The same C-terminal domain is also required for binding of a cellular 48-kDa phosphoprotein, C-terminal binding protein (CtBP). We have cloned the cDNA for CtBP via yeast two-hybrid interaction cloning. The cDNA encodes a 439-amino acid (48 kDa) protein that specifically interacts with exon 2 in yeast two-hybrid, in vitro protein binding, and in vivo coimmunoprecipitation analyses. This protein requires residues 225-238 of the 243R E1A protein for interaction. The predicted protein sequence of the isolated cDNA is identical to amino acid sequences obtained from peptides prepared from biochemically purified CtBP. Fine mapping of the CtBP-binding domain revealed that a 6-amino acid motif highly conserved among the E1A proteins of various human and animal adenoviruses is required for this interaction. These results suggest that interaction of CtBP with the E1A proteins may play a critical role in adenovirus replication and oncogenic transformation. PMID- 7479822 TI - Specific and high-affinity binding of inositol phosphates to an isolated pleckstrin homology domain. AB - Pleckstrin homology (PH) domains are found in many signaling molecules and are thought to be involved in specific intermolecular interactions. Their binding to several proteins and to membranes containing 1-alpha-phosphatidylinositol 4,5 bisphosphate [PtdIns(4,5)P2] has been reported. A region that includes the PH domain has also been implicated in binding of phospholipase C-delta 1 (PLC-delta 1) to both PtdIns(4,5)P2 and D-myo-inositol 1,4,5-trisphosphate [Ins(1,4,5)P3] [Cifuentes, M. E., Delaney, T. & Rebecchi, M. J. (1994) J. Biol. Chem. 269, 1945 1948]. We report herein that the isolated PH domain from PLC-delta 1 binds to both PtdIns(4,5)P2 and Ins(1,4,5)P3 with high affinity and shows the same binding specificity seen by others with whole PLC-delta 1. Thus the PH domain is functionally and structurally modular. These results demonstrate stereo-specific high-affinity binding by an isolated PH domain and further support a functional role for PH domains in the regulation of PLC isoforms. Other PH domains did not bind strongly to the compounds tested, suggesting that inositol phosphates and phospholipids are not likely physiological ligands for all PH domains. Nonetheless, since all PH-domain-containing proteins are associated with membrane surfaces, several PH domains bind to specific sites on membranes, and PH domains appear to be electrostatically polarized, a possible general role for PH domains in membrane association is suggested. PMID- 7479823 TI - Identification and characterization of a Drosophila nuclear receptor with the ability to inhibit the ecdysone response. AB - In a search for retinoid X receptor-like molecules in Drosophila, we have identified an additional member of the nuclear receptor superfamily, XR78E/F. In the DNA-binding domain, XR78E/F is closely related to the mammalian receptor TR2, as well as to the nuclear receptors Coup-TF and Seven-up. We demonstrate that XR78E/F binds as a homodimer to direct repeats of the sequence AGGTCA. In transient transfection assays, XR78E/F represses ecdysone signaling in a DNA binding-dependent fashion. XR78E/F has its highest expression in third-instar larvae and prepupae. These experiments suggest that XR78E/F may play a regulatory role in the transcriptional cascade triggered by the hormone ecdysone in Drosophila. PMID- 7479824 TI - Indirect coupling of phosphate release to de novo tension generation during muscle contraction. AB - A key question in muscle contraction is how tension generation is coupled to the chemistry of the actomyosin ATPase. Biochemical and mechanochemical experiments link tension generation to a change in structure associated with phosphate release. Length-jump and temperature-jump experiments, on the other hand, implicate phase 2slow, a significantly faster, markedly strain-sensitive kinetic process in tension generation. We use a laser temperature jump to probe the kinetics and mechanism of tension generation in skinned rabbit psoas fibers--an appropriate method since both phosphate release and phase 2slow are readily perturbed by temperature. Kinetics characteristic of the structural change associated with phosphate release are observed only when phosphate is added to fibers. When present, it causes a reduction in fiber tension; otherwise, no force is generated when it is perturbed. We therefore exclude this step from tension generation. The kinetics of de novo tension generation by the temperature-jump equivalent of phase 2slow appear unaffected by phosphate binding. We therefore propose that phosphate release is indirectly coupled to de novo tension generation via a steady-state flux through an irreversible step. We conclude that tension generation occurs in the absence of chemical change as the result of an entropy-driven transition between strongly bound crossbridges in the actomyosin ADP state. The mechanism resembles the operation of a clock, with phosphate release providing the energy to tension the spring, and the irreversible step functions as the escapement mechanism, which is followed in turn by tension generation as the movement of the hands. PMID- 7479825 TI - Expression and signaling specificity of the IFNAR chain of the type I interferon receptor complex. AB - The IFNAR chain of the type I interferon (IFN) receptor (IFNIR) undergoes rapid ligand-dependent tyrosine phosphorylation and acts as a species-specific transducer for type I IFN action. Using the vaccinia/T7 expression system to amplify IFNAR expression, we found that human HeLa-S3 cells transiently express high levels of cell surface IFNAR chains (approximately 250,000 chains per cell). Metabolic labeling and immunoblot analysis of transfected HeLa cells show that the IFNAR chain is initially detected as 65-kDa and 98-kDa precursors, and then as the 130-kDa mature protein. Due to variation in N-glycosylation, the apparent molecular mass of the mature IFNAR chain varies from 105 to 135 kDa in different cells. IFNIR structure was characterized in various human cell lines by analyzing 125I-labeled IFN cross-linked complexes recognized by various antibodies against IFNIR subunits and JAK protein-tyrosine kinases. Precipitation of cross-linked material from Daudi cells with anti-IFNAR antibodies showed that IFNAR was present in a 240-kDa complex. Precipitation of cross-linked material from U937 cells with anti-TYK2 sera revealed a 240-kDa complex, which apparently did not contain IFNAR and was not present in IFN-resistant HEC1B cells. The tyrosine phosphorylation and down-regulation of the IFNAR chain were induced by type I IFN in several human cell lines of diverse origins but not in HEC1B cells. However, of type I IFNs, IFN-beta uniquely induced the tyrosine phosphorylation of a 105 kDa protein associated with the IFNAR chain in two lymphoblastoid cell lines (Daudi and U266), demonstrating the specificity of transmembrane signaling for IFN-beta and IFN-alpha through the IFNAR chain. PMID- 7479826 TI - Passive immunotherapy for retroviral disease: influence of major histocompatibility complex type and T-cell responsiveness. AB - Administration of virus-specific antibodies is known to be an effective early treatment for some viral infections. Such immunotherapy probably acts by antibody mediated neutralization of viral infectivity and is often thought to function independently of T-cell-mediated immune responses. In the present experiments, we studied passive antibody therapy using Friend murine leukemia virus complex as a model for an immunosuppressive retroviral disease in adult mice. The results showed that antibody therapy could induce recovery from a well-established retroviral infection. However, the success of therapy was dependent on the presence of both CD4+ and CD8+ T lymphocytes. Thus, cell-mediated responses were required for recovery from infection even in the presence of therapeutic levels of antibody. The major histocompatibility type of the mice was also an important factor determining the relative success of antibody therapy in this system, but it was less critical for low-dose than for high-dose infections. Our results imply that limited T-cell responsiveness as dictated by major histocompatibility genes and/or stage of disease may have contributed to previous immunotherapy failures in AIDS patients. Possible strategies to improve the efficacy of future therapies are discussed. PMID- 7479827 TI - Molecular basis of human mitochondrial very-long-chain acyl-CoA dehydrogenase deficiency causing cardiomyopathy and sudden death in childhood. AB - beta-Oxidation of long-chain fatty acids provides the major source of energy in the heart. Defects in enzymes of the beta-oxidation pathway cause sudden, unexplained death in childhood, acute hepatic encephalopathy or liver failure, skeletal myopathy, and cardiomyopathy. Very-long-chain acyl-CoA dehydrogenase [VLCAD; very-long-chain-acyl-CoA:(acceptor) 2,3-oxidoreductase, EC 1.3.99.13] catalyzes the first step in beta-oxidation. We have isolated the human VLCAD cDNA and gene and determined the complete nucleotide sequences. Polymerase chain reaction amplification of VLCAD mRNA and genomic exons defined the molecular defects in two patients with VLCAD deficiency who presented with unexplained cardiac arrest and cardiomyopathy. In one, a homozygous mutation in the consensus dinucleotide of the donor splice site (g+1-->a) was associated with universal skipping of the prior exon (exon 11). The second patient was a compound heterozygote, with a missense mutation, C1837-->T, changing the arginine at residue 613 to tryptophan on one allele and a single base deletion at the intron exon 6 boundary as the second mutation. This initial delineation of human mutations in VLCAD suggests that VLCAD deficiency reduces myocardial fatty acid beta-oxidation and energy production and is associated with cardiomyopathy and sudden death in childhood. PMID- 7479828 TI - Fasciclin II controls proneural gene expression in Drosophila. AB - Fasciclin II (Fas II), an NCAM-like cell adhesion molecule in Drosophila, is expressed on a subset of embryonic axons and controls selective axon fasiculation. Fas II is also expressed in imaginal discs. Here we use genetic analysis to show that Fas II is required for the control of proneural gene expression. Clusters of cells in the eye-antennal imaginal disc express the achaete proneural gene and give rise to mechanosensory neurons; other clusters of cells express the atonal gene and give rise to ocellar photoreceptor neurons. In fasII loss-of-function mutants, the expression of both proneural genes is absent in certain locations, and, as a result, the corresponding sensory precursors fail to develop. In fasII gain-of-function conditions, extra sensory structures arise from this same region of the imaginal disc. Mutations in the Abelson tyrosine kinase gene show dominant interactions with fasII mutations, suggesting that Abl and Fas II function in a signaling pathway that controls proneural gene expression. PMID- 7479829 TI - Calicheamicin-DNA complexes: warhead alignment and saccharide recognition of the minor groove. AB - The solution structures of calicheamicin gamma 1I, its cycloaromatized analog (calicheamicin epsilon), and its aryl tetrasaccharide complexed to a common DNA hairpin duplex have been determined by NMR and distance-refined molecular dynamics computations. Sequence specificity is associated with carbohydrate-DNA recognition that places the aryl tetrasaccharide component of all three ligands in similar orientations in the minor groove at the d(T-C-C-T).d(A-G-G-A) segment. The complementary fit of the ligands and the DNA minor groove binding site creates numerous van der Waals contacts as well as hydrogen bonding interactions. Notable are the iodine and sulfur atoms of calicheamicin that hydrogen bond with the exposed amino proton of the 5'- and 3'-guanines, respectively, of the d(A-G-G A) segment. The sequence-specific carbohydrate binding orients the enediyne aglycone of calicheamicin gamma 1I such that its C3 and C6 proradical centers are adjacent to the cleavage sites. While the enediyne aglycone of calicheamicin gamma 1I is tilted relative to the helix axis and spans the minor groove, the cycloaromatized aglycone is aligned approximately parallel to the helix axis in the respective complexes. Specific localized conformational perturbations in the DNA have been identified from imino proton complexation shifts and changes in specific sugar pucker patterns on complex formation. The helical parameters for the carbohydrate binding site are comparable with corresponding values in B-DNA fibers while a widening of the groove is observed at the adjacent aglycone binding site. PMID- 7479830 TI - A single-stranded DNA binding protein binds the origin of replication of the duplex kinetoplast DNA. AB - Replication of the kinetoplast DNA (kDNA) minicircle of trypanosomatids initiates at a conserved 12-nt sequence, 5'-GGGGTTGGTGTA-3', termed the universal minicircle sequence (UMS). A sequence-specific single-stranded DNA-binding protein from Crithidia fasciculata binds the heavy strand of the 12-mer UMS. Whereas this UMS-binding protein (UMSBP) does not bind a duplex UMS dodecamer, it binds the double-stranded kDNA minicircle as well as a duplex minicircle fragment containing the origin-associated UMS. Binding of the minicircle origin region by the single-stranded DNA binding protein suggested the local unwinding of the DNA double helix at this site. Modification of thymine residues at this site by KMnO4 revealed that the UMS resides within an unwound or otherwise sharply distorted DNA at the minicircle origin region. Computer analysis predicts the sequence directed curving of the minicircle origin region. Electrophoresis of a minicircle fragment containing the origin region in polyacrylamide gels revealed a significantly lower electrophoretic mobility than expected from its length. The fragment anomalous electrophoretic mobility is displayed only in its native conformation and is dependent on temperature and gel porosity, indicating the local curving of the DNA double helix. We suggest that binding of UMSBP at the minicircle origin of replication is possible through local unwinding of the DNA double helix at the UMS site. It is hypothesized here that this local melting is initiated through the untwisting of unstacked dinucleotide sequences at the bent origin site. PMID- 7479831 TI - Association of a M(r) 90,000 phosphoprotein with protein kinase PKR in cells exhibiting enhanced phosphorylation of translation initiation factor eIF-2 alpha and premature shutoff of protein synthesis after infection with gamma 134.5- mutants of herpes simplex virus 1. AB - The protein encoded by the gamma 134.5 gene of herpes simplex virus precludes premature shutoff of protein synthesis in human cells triggered by stress associated with onset of viral DNA synthesis. The carboxyl terminus of the protein is essential for this function. This report indicates that the shutoff of protein synthesis is not due to mRNA degration because mRNA from wild-type or gamma 134.5- virus-infected cells directs protein synthesis. Analyses of the posttranslational modifications of translation initiation factor eIF-2 showed the following: (i) eIF-2 alpha was selectively phosphorylated by a kinase present in ribosome-enriched fraction of cells infected with gamma 134.5- virus. (ii) Endogenous eIF-2 alpha was totally phosphorylated in cells infected with gamma 134.5- virus or a virus lacking the 3' coding domain of the gamma 134.5 gene but was not phosphorylated in mock-infected or wild-type virus-infected cells. (iii) Immune precipitates of the PKR kinase that is responsible for regulation of protein synthesis of some cells by phosphorylation of eIF-2 alpha yielded several phosphorylated polypeptides. Of particular significance were two observations. First, phosphorylation of PKR kinase was elevated in all infected cells relative to the levels in mock-infected cells. Second, the precipitates from lysates of cells infected with gamma 134.5- virus or a virus lacking the 3' coding domain of the gamma 134.5 gene contained an additional labeled phosphoprotein of M(r) 90,000 (p90). This phosphoprotein was present in only trace amounts in the immunoprecipitate from cells infected with wild-type virus or mutants lacking a portion of the 5' domain of gamma 134.5. We conclude that in the absence of gamma 134.5 protein, PKR kinase complexes with the p90 phosphoprotein and shuts off protein synthesis by phosphorylation of the alpha subunit of translation initiation factor eIF-2. PMID- 7479832 TI - Multiple protein kinase A-regulated events are required for transcriptional induction by cAMP. AB - The second messenger cAMP stimulates the expression of numerous genes via the protein kinase A-mediated phosphorylation of the cAMP response element-binding protein (CREB) at Ser-133. Ser-133 phosphorylation, in turn, appears to induce target gene expression by promoting interaction between CREB and CBP, a 265-kDa nuclear phospho-CREB-binding protein. It is unclear, however, whether Ser-133 phosphorylation per se is sufficient for CREB-CBP complex formation and for target gene induction in vivo. Here we examine CREB activity in Jurkat T cells after stimulation of the T-cell receptor (TCR), an event that leads to calcium entry and diacylglycerol production. Triggering of the TCR stimulated Ser-133 phosphorylation of CREB with high stoichiometry, but TCR activation did not promote CREB-CBP complex formation or target gene induction unless suboptimal doses of cAMP agonist were provided as a costimulus. Our results demonstrate that, in addition to mediating Ser-133 phosphorylation of CREB, protein kinase A regulates additional proteins that are required for recruitment of the transcriptional apparatus to cAMP-responsive genes. PMID- 7479833 TI - The nuclear matrix protein NMP-1 is the transcription factor YY1. AB - NMP-1 was initially identified as a nuclear matrix-associated DNA-binding factor that exhibits sequence-specific recognition for the site IV regulatory element of a histone H4 gene. This distal promoter domain is a nuclear matrix interaction site. In the present study, we show that NMP-1 is the multifunctional transcription factor YY1. Gel-shift and Western blot analyses demonstrate that NMP-1 is immunoreactive with YY1 antibody. Furthermore, purified YY1 protein specifically recognizes site IV and reconstitutes the NMP-1 complex. Western blot and gel-shift analyses indicate that YY1 is present within the nuclear matrix. In situ immunofluorescence studies show that a significant fraction of YY1 is localized in the nuclear matrix, principally but not exclusively associated with residual nucleoli. Our results confirm that NMP-1/YY1 is a ubiquitous protein that is present in both human cells and in rat osteosarcoma ROS 17/2.8 cells. The finding that NMP-1 is identical to YY1 suggests that this transcriptional regulator may mediate gene-matrix interactions. Our results are consistent with the concept that the nuclear matrix may functionally compartmentalize the eukaryotic nucleus to support regulation of gene expression. PMID- 7479834 TI - Mxi2, a mitogen-activated protein kinase that recognizes and phosphorylates Max protein. AB - We describe Mxi2, a human protein that interacts with Max protein, the heterodimeric partner of the Myc oncoprotein. Mxi2 encodes a 297-residue protein whose sequence indicates that it is related to extracellular signal-regulated kinases (ERK protein kinases). Mxi2 in yeast interacts with Max and with the C terminus of c-Myc. Mxi2 phosphorylates Max both in vitro and in vivo. The Mxi2 putative substrate recognition region has sequence similarity to the helix-loop helix region in Max and c-Myc, suggesting that substrate recognition might be mediated via this motif. Phosphorylation by Mxi2 may affect the ability of Max to oligomerize with itself and its partners, bind DNA, or regulate gene expression. PMID- 7479835 TI - Electrophoretic variation in adenylate kinase of Neisseria meningitidis is due to inter- and intraspecies recombination. AB - In prokaryotic and eukaryotic organisms, the electrophoretic variation in housekeeping enzymes from natural populations is assumed to have arisen by the accumulation of stochastic predominantly neutral mutations. In the naturally transformable bacterium Neisseria meningitidis, we show that variation in the electrophoretic mobility of adenylate kinase is due to inter- and intraspecies recombination rather than mutation. The nucleotide sequences of the adenylate kinase gene (adk) from isolates that express the predominant slow electrophoretic variant were rather uniform, differing in sequence at an average of 1.1% of nucleotide sites. The adk sequences of rare isolates expressing the fast migrating variant were identical to each other but had a striking mosaic structure when compared to the adk genes from strains expressing the predominant variant. Thus the sequence from the fast variants was identical to those of typical slow variants in the first 158 bp of the gene but differed by 8.4% in the rest of the gene (nt 159-636). The fast electrophoretic variant appears to have arisen by the replacement of most of the meningococcal gene with the corresponding region from the adk gene of a closely related Neisseria species. The adk genes expressing the electrophoretic variant with intermediate mobility were perfect, or almost perfect, recombinants between the adk genes expressing the fast and slow variants. Recombination may, therefore, play a major role in the generation of electrophoretically detectable variation in housekeeping enzymes of some bacterial species. PMID- 7479838 TI - The ability to associate with activation domains in vitro is not required for the TATA box-binding protein to support activated transcription in vivo. AB - The TATA box-binding protein (TBP) interacts in vitro with the activation domains of many viral and cellular transcription factors and has been proposed to be a direct target for transcriptional activators. We have examined the functional relevance of activator-TBP association in vitro to transcriptional activation in vivo. We show that alanine substitution mutations in a single loop of TBP can disrupt its association in vitro with the activation domains of the herpes simplex virus activator VP16 and of the human tumor suppressor protein p53; these mutations do not, however, disrupt the transcriptional response of TBP to either activation domain in vivo. Moreover, we show that a region of VP16 distinct from its activation domain can also tightly associate with TBP in vitro, but fails to activate transcription in vivo. These data suggest that the ability of TBP to interact with activation domains in vitro is not directly relevant to its ability to support activated transcription in vivo. PMID- 7479837 TI - A behavioral screen for isolating zebrafish mutants with visual system defects. AB - Optokinetic and phototactic behaviors of zebrafish larvae were examined for their usefulness in screening for recessive defects in the visual system. The optokinetic response can be reliably and rapidly detected in 5-day larvae, whereas the phototactic response of larvae is variable and not robust enough to be useful for screening. We therefore measured optokinetic responses of mutagenized larvae as a genetic screen for visual system defects. Third generation larvae, representing 266 mutagenized genomes, were examined for abnormal optokinetic responses. Eighteen optokinetic-defective mutants were identified and two mutants that did not show obvious morphological defects, no optokinetic response a (noa) and partial optokinetic response a (poa), were studied further. We recorded the electroretinogram (ERG) to determine whether these two mutations affect the retina. The b-wave of noa larvae was grossly abnormal, being delayed in onset and significantly reduced in amplitude. In contrast, the ERG waveform of poa larvae was normal, although the b-wave was reduced in amplitude in bright light. Histologically, the retinas of noa and poa larvae appeared normal. We conclude that noa larvae have a functional defect in the outer retina, whereas the outer retina of poa larvae is likely to be normal. PMID- 7479836 TI - Ultra-low concentrations of naloxone selectively antagonize excitatory effects of morphine on sensory neurons, thereby increasing its antinociceptive potency and attenuating tolerance/dependence during chronic cotreatment. AB - Ultra-low picomolar concentrations of the opioid antagonists naloxone (NLX) and naltrexone (NTX) have remarkably potent antagonist actions on excitatory opioid receptor functions in mouse dorsal root ganglion (DRG) neurons, whereas higher nanomolar concentrations antagonize excitatory and inhibitory opioid functions. Pretreatment of naive nociceptive types of DRG neurons with picomolar concentrations of either antagonist blocks excitatory prolongation of the Ca(2+) dependent component of the action potential duration (APD) elicited by picomolar nanomolar morphine and unmasks inhibitory APD shortening. The present study provides a cellular mechanism to account for previous reports that low doses of NLX and NTX paradoxically enhance, instead of attenuate, the analgesic effects of morphine and other opioid agonists. Furthermore, chronic cotreatment of DRG neurons with micromolar morphine plus picomolar NLX or NTX prevents the development of (i) tolerance to the inhibitory APD-shortening effects of high concentrations of morphine and (ii) supersensitivity to the excitatory APD prolonging effects of nanomolar NLX as well as of ultra-low (femtomolar picomolar) concentrations of morphine and other opioid agonists. These in vitro studies suggested that ultra-low doses of NLX or NTX that selectively block the excitatory effects of morphine may not only enhance the analgesic potency of morphine and other bimodally acting opioid agonists but also markedly attenuate their dependence liability. Subsequent correlative studies have now demonstrated that cotreatment of mice with morphine plus ultra-low-dose NTX does, in fact, enhance the antinociceptive potency of morphine in tail-flick assays and attenuate development of withdrawal symptoms in chronic, as well as acute, physical dependence assays. PMID- 7479839 TI - Genetic and comparative analyses reveal an alternative secondary structure in the region of nt 912 of Escherichia coli 16S rRNA. AB - Mutations at position 912 of Escherichia coli 16S rRNA result in two notable phenotypes. The C-->U transition confers resistance to streptomycin, a translational-error-inducing antibiotic, while a C-->G transversion causes marked retardation of cell growth rate. Starting with the slow-growing G912 mutant, random mutagenesis was used to isolate a second site mutation that restored growth nearly to the wild-type rate. The second site mutation was identified as a G-->C transversion at position 885 in 16S rRNA. Cells containing the G912 mutation had an increased doubling time, abnormal sucrose gradient ribosome/subunit profile, increased sensitivity to spectinomycin, dependence upon streptomycin for growth in the presence of spectinomycin, and slower translation rate, whereas cells with the G912/C885 double mutation were similar to wild type in these assays. Comparative analysis showed there was significant covariation between positions 912 and 885. Thus the second-site suppressor analysis, the functional assays, and the comparative data suggest that the interaction between nt 912 and nt 885 is conserved and necessary for normal ribosome function. Furthermore, the comparative data suggest that the interaction extends to include G885-G886-G887 pairing with C912-U911-C910. An alternative secondary structure element for the central domain of 16S rRNA is proposed. PMID- 7479840 TI - Identification of a point mutation in the catalytic domain of the protooncogene c kit in peripheral blood mononuclear cells of patients who have mastocytosis with an associated hematologic disorder. AB - Both stem cells and mast cells express c-kit and proliferate after exposure to c kit ligand. Mutations in c-kit may enhance or interfere with the ability of c-kit receptor to initiate the intracellular pathways resulting in cell proliferation. These observations suggested to us that mastocytosis might in some patients result from mutations in c-kit. cDNA synthesized from peripheral blood mononuclear cells of patients with indolent mastocytosis, mastocytosis with an associated hematologic disorder, aggressive mastocytosis, solitary mastocytoma, and chronic myelomonocytic leukemia unassociated with mastocytosis was thus screened for a mutation of c-kit. This analysis revealed that four of four mastocytosis patients with an associated hematologic disorder with predominantly myelodysplastic features had an A-->T substitution at nt 2468 of c-kit mRNA that causes an Asp-816-->Val substitution. One of one patient examined who had mastocytosis with an associated hematologic disorder had the corresponding mutation in genomic DNA. Identical or similar amino acid substitutions in mast cell lines result in ligand-independent autophosphorylation of the c-kit receptor. This mutation was not identified in the patients within the other disease categories or in 67 of 67 controls. The identification of the point mutation Asp816Val in c-kit in patients with mastocytosis with an associated hematologic disorder provides insight not only into the pathogenesis of this form of mastocytosis but also into how hematopoiesis may become dysregulated and may serve to provide a means of confirming the diagnosis, assessing prognosis, and developing intervention strategies. PMID- 7479841 TI - Redefining the Epstein-Barr virus-encoded nuclear antigen EBNA-1 gene promoter and transcription initiation site in group I Burkitt lymphoma cell lines. AB - The Epstein-Barr virus-encoded nuclear antigen EBNA-1 gene promoter for the restricted Epstein-Barr virus (EBV) latency program operating in group I Burkitt lymphoma (BL) cell lines was previously identified incorrectly. Here we present evidence from RACE (rapid amplification of cDNA ends) cloning, reverse transcription-PCR, and S1 nuclease analyses, which demonstrates that the EBNA-1 gene promoter in group I BL cell lines is located in the viral BamHI Q fragment, immediately upstream of two low-affinity EBNA-1 binding sites. Transcripts initiated from this promoter, referred to as Qp, have the previously reported Q/U/K exon splicing pattern. Qp is active in group I BL cell lines but not in group III BL cell lines or in EBV immortalized B-lymphoblastoid cell lines. In addition, transient transfection of Qp-driven reporter constructs into both an EBV-negative BL cell line and a group I BL cell line gave rise to correctly initiated transcripts. Inspection of Qp revealed that it is a TATA-less promoter whose architecture is similar to the promoters of housekeeping genes, suggesting that Qp may be a default promoter which ensures EBNA-1 expression in cells that cannot run the full viral latency program. Elucidation of the genetic mechanism responsible for the EBNA-1-restricted program of EBV latency is an essential step in understanding control of viral latency in EBV-associated tumors. PMID- 7479842 TI - Attenuated virulence of chitin-deficient mutants of Candida albicans. AB - We have analyzed the role of chitin, a cell-wall polysaccharide, in the virulence of Candida albicans. Mutants with a 5-fold reduction in chitin were obtained in two ways: (i) by selecting mutants resistant to Calcofluor, a fluorescent dye that binds to chitin and inhibits growth, and (ii) by disrupting CHS3, the C. albicans homolog of CSD2/CAL1/DIT101/KT12, a Saccharomyces cerevisiae gene required for synthesis of approximately 90% of the cell-wall chitin. Chitin deficient mutants have no obvious alterations in growth rate, sugar assimilation, chlamydospore formation, or germ-tube formation in various media. When growing vegetatively in liquid media, the mutants tend to clump and display minor changes in morphology. Staining of cells with the fluorescent dye Calcofluor indicates that CHS3 is required for synthesis of the chitin rings found on the surface of yeast cells but not formation of septa in either yeast cells or germ tubes. Despite their relatively normal growth, the mutants are significantly less virulent than the parental strain in both immunocompetent and immunosuppressed mice; at 13 days after infection, survival was 95% in immunocompetent mice that received chs3/chs3 cells and 10% in immunocompetent mice that received an equal dose of chs3/CHS3 cells. Chitin-deficient strains can colonize the organs of infected mice, suggesting that the reduced virulence of the mutants is not due to accelerated clearing. PMID- 7479843 TI - Potent, structurally constrained agonists and competitive antagonists of corticotropin-releasing factor. AB - Predictive methods, physicochemical measurements, and structure activity relationship studies suggest that corticotropin-releasing factor (CRF; corticoliberin), its family members, and competitive antagonists (resulting from N-terminal deletions) usually assume an alpha-helical conformation when interacting with the CRF receptor(s). To test this hypothesis further, we have scanned the whole sequence of the CRF antagonist [D-Phe12,Nle21,38]r/hCRF-(12-41) (r/hCRF, rat/human CRF; Nle, norleucine) with an i-(i + 3) bridge consisting of the Glu-Xaa-Xaa-Lys scaffold. We have found astressin [cyclo(30-33)[D Phe12,Nle21,38,Glu30,Lys33]r/ hCRF(12-41)] to be approximately 30 times more potent than [D-Phe12,Nle21,38]r/hCRF-(12-41), our present standard, and 300 times more potent than the corresponding linear analog in an in vitro pituitary cell culture assay. Astressin has low affinity for the CRF binding protein and high affinity (Ki = 2 nM) for the cloned pituitary receptor. Radioiodinated [D-125I Tyr12]astressin was found to be a reliable ligand for binding assays. In vivo, astressin is significantly more potent than any previously tested antagonist in reducing hypophyseal corticotropin (ACTH) secretion in stressed or adrenalectomized rats. The cyclo(30-33)[Ac-Pro4,D-Phe12,Nle21,38,Glu30,Lys33++ +]r/hCRF-(4-41) agonist and its linear analog are nearly equipotent, while the antagonist astressin and its linear form vary greatly in their potencies. This suggests that the lactam cyclization reinstates a structural constraint in the antagonists that is normally induced by the N terminus of the agonist. PMID- 7479844 TI - Characterization of Arabidopsis thaliana cDNAs that render yeasts tolerant toward the thiol-oxidizing drug diamide. AB - Diamide oxidizes cellular thiols and induces oxidative stress. To isolate plant genes which may, when overexpressed, increase tolerance of plants toward oxidative damage, an in vivo diamide tolerance screening in yeasts was used. An Arabidopsis cDNA library in a yeast expression vector was used to transform a yeast strain with intact antioxidant defense. Cells from approximately 10(5) primary transformants were selected for resistance to diamide. Three Arabidopsis cDNAs which confer diamide tolerance were isolated. This drug tolerance was specific and no cross tolerance toward hydroperoxides was found. One cDNA (D3) encodes a polypeptide which has an amino-terminal J domain characteristic of a divergent family of DnaJ chaperones. Another (D18) encodes a putative dTDP-D glucose 4,6-dehydratase. Surprisingly, the third cDNA (D22) encodes a plant homolog of gamma-glutamyltransferases. It would have been difficult to predict that the expression of those genes would lead to an improved survival under conditions of depletion of cellular thiols. Hence, we suggest that this cloning approach may be a useful contribution to the isolation of plant genes that can help to cope with oxidative stress. PMID- 7479845 TI - Complexes of p21RAS with JUN N-terminal kinase and JUN proteins. AB - RAS gene-encoded p21 protein has been found to increase in vitro phosphorylation of JUN via its kinase, JUN N-terminal kinase (JNK). This effect is mediated by increased phosphorylation of JNK in the presence of wild-type and oncogenic (Val 12) p21 protein in a dose-dependent manner. Oncogenic p21 protein is more potent in mediating this effect than its normal counterpart. Both normal and oncogenic p21 proteins bind to purified JNK and to JNK that is present in cell extracts from transformed fibroblasts and melanoma cells. Oncogenic and normal p21 proteins have also been found to bind to bacterially expressed JUN protein. This binding is dose dependent, enhanced by the presence of GTP, and depends on the presence of the first 89 amino acids of JUN (the delta domain), as it does not occur with v-jun. While the ability of both normal and oncogenic p21 proteins to bind JNK is strongly inhibited by a p21 peptide corresponding to aa 96-110, and more weakly inhibited by the p21 peptide corresponding to aa 115-126, p21-JUN interaction is inhibited by peptides corresponding to aa 96-110 and, to a lesser degree, by peptides corresponding to aa 35-47. The results suggest that the p21 protein interacts specifically with both JNK and JUN proteins. PMID- 7479846 TI - The versican C-type lectin domain recognizes the adhesion protein tenascin-R. AB - The core proteins of large chondroitin sulfate proteoglycans contain a C-type lectin domain. The lectin domain of one of these proteoglycans, versican, was expressed as a recombinant 15-kDa protein and shown to bind to insolubilized fucose and GlcNAc. The lectin domain showed strong binding in a gel blotting assay to a glycoprotein doublet in rat brain extracts. The binding was calcium dependent and abolished by chemical deglycosylation treatment of the ligand glycoprotein. The versican-binding glycoprotein was identified as the cell adhesion protein tenascin-R, and versican and tenascin-R were both found to be localized in the granular layer of rat cerebellum. These results show that the versican lectin domain is a binding domain with a highly targeted specificity. It may allow versican to assemble complexes containing proteoglycan, an adhesion protein, and hyaluronan. PMID- 7479847 TI - Location of the active site of allosteric chorismate mutase from Saccharomyces cerevisiae, and comments on the catalytic and regulatory mechanisms. AB - The active site of the allosteric chorismate mutase (chorismate pyruvatemutase, EC 5.4.99.5) from yeast Saccharomyces cerevisiae (YCM) was located by comparison with the mutase domain (ECM) of chorismate mutase/prephenate dehydratase [prephenate hydro-lyase (decarboxylating), EC 4.2.1.51] (the P protein) from Escherichia coli. Active site domains of these two enzymes show very similar four helix bundles, each of 94 residues which superimpose with a rms deviation of 1.06 A. Of the seven active site residues, four are conserved: the two arginines, which bind to the inhibitor's two carboxylates; the lysine, which binds to the ether oxygen; and the glutamate, which binds to the inhibitor's hydroxyl group in ECM and presumably in YCM. The other three residues in YCM (ECM) are Thr-242 (Ser 84), Asn-194 (Asp-48), and Glu-246 (Gln-88). This Glu-246, modeled close to the ether oxygen of chorismate in YCM, may function as a polarizing or ionizable group, which provides another facet to the catalytic mechanism. PMID- 7479849 TI - Isolation, regulation, and DNA-binding properties of three Drosophila nuclear hormone receptor superfamily members. AB - We have designed a rapid cloning and screening strategy to identify new members of the nuclear hormone receptor superfamily that are expressed during the onset of Drosophila metamorphosis. Using this approach, we isolated three Drosophila genes, designated DHR38, DHR78, and DHR96. All three genes are expressed throughout third-instar larval and prepupal development. DHR38 is the Drosophila homolog of NGFI-B and binds specifically to an NGFI-B response element. DHR78 and DHR96 are orphan receptor genes. DHR78 is induced by 20-hydroxyecdysone (20E) in cultured larval organs, and its encoded protein binds to two AGGTCA half-sites arranged as either direct or palindromic repeats. DHR96 is also 20E-inducible, and its encoded protein binds selectively to the hsp27 20E response element. The 20E receptor can bind to each of the sequences recognized by DHR78 and DHR96, indicating that these proteins may compete with the receptor for binding to a common set of target sequences. PMID- 7479848 TI - Stimulation-dependent I kappa B alpha phosphorylation marks the NF-kappa B inhibitor for degradation via the ubiquitin-proteasome pathway. AB - The nuclear translocation of NF-kappa B follows the degradation of its inhibitor, I kappa B alpha, an event coupled with stimulation-dependent inhibitor phosphorylation. Prevention of the stimulation-dependent phosphorylation of I kappa B alpha, either by treating cells with various reagents or by mutagenesis of certain putative I kappa B alpha phosphorylation sites, abolishes the inducible degradation of I kappa B alpha. Yet, the mechanism coupling the stimulation-induced phosphorylation with the degradation has not been resolved. Recent reports suggest a role for the proteasome in I kappa B alpha degradation, but the mode of substrate recognition and the involvement of ubiquitin conjugation as a targeting signal have not been addressed. We show that of the two forms of I kappa B alpha recovered from stimulated cells in a complex with RelA and p50, only the newly phosphorylated form, pI kappa B alpha, is a substrate for an in vitro reconstituted ubiquitin-proteasome system. Proteolysis requires ATP, ubiquitin, a specific ubiquitin-conjugating enzyme, and other ubiquitin-proteasome components. In vivo, inducible I kappa B alpha degradation requires a functional ubiquitin-activating enzyme and is associated with the appearance of high molecular weight adducts of I kappa B alpha. Ubiquitin mediated protein degradation may, therefore, constitute an integral step of a signal transduction process. PMID- 7479851 TI - Delayed male maturity is a cost of producing large sperm in Drosophila. AB - Among fruit-fly species of the genus Drosophila there is remarkable variation in sperm length, with some species producing gigantic sperm (e.g., > 10 times total male body length). These flies are also unusual in that males of some species exhibit a prolonged adult nonreproductive phase. We document sperm length, body size, and sex-specific ages of reproductive maturity for 42 species of Drosophila and, after controlling for phylogeny, test hypotheses to explain the variation in rates of sexual maturation. Results suggest that delayed male maturity is a cost of producing long sperm. A possible physiological mechanism to explain the observed relationship is discussed. PMID- 7479850 TI - Alignment of a 1.2-Mb chromosomal region from three strains of Rhodobacter capsulatus reveals a significantly mosaic structure. AB - High-resolution physical maps of the genomes of three Rhodobacter capsulatus strains, derived from ordered cosmid libraries, were aligned. The 1.2-Mb segment of the SB1003 genome studied here is adjacent to a 1-Mb region analyzed previously [Fonstein, M., Nikolskaya, T. & Haselkorn, H. (1995) J. Bacteriol. 177, 2368-2372]. Probes derived from the ordered cosmid set of R. capsulatus SB1003 were used to link cosmids from the St. Louis and 2.3.1 strain libraries. Cosmids selected this way did not merge into a single contig but formed several unlinked groups. EcoRV restriction maps of the ordered cosmids were then constructed using lambda terminase and fused to derive fragments of the chromosomal map. In order to link these fragments, their ends were transcribed to produce secondary probes for hybridization to gridded cosmid libraries of the same strains. This linking reduced the number of subcontigs to three for the St. Louis strain and one for the 2.3.1 strain. Hybridization of the same probes back to the ordered cosmid set of SB1003 positioned the subcontigs on the high resolution physical map of SB1003. The final alignment of the restriction maps shows numerous large and small translocations in this 1.2-Mb chromosomal region of the three Rhodobacter strains. In addition, the chromosomes of the three strains, whose fine-structure maps can now be compared over 2.2 Mb, are seen to contain regions of 15-80 kb in which restriction sites are highly polymorphic, interspersed among regions in which the positions of restriction sites are highly conserved. PMID- 7479852 TI - Serine phosphorylation of human P450c17 increases 17,20-lyase activity: implications for adrenarche and the polycystic ovary syndrome. AB - Microsomal cytochrome P450c17 catalyzes both steroid 17 alpha-hydroxylase activity and scission of the C17-C20 steroid bond (17,20-lyase) on the same active site. Adrenal 17 alpha-hydroxylase activity is needed to produce cortisol throughout life, but 17,20-lyase activity appears to be controlled independently in a complex, age-dependent pattern. We show that human P450c17 is phosphorylated on serine and threonine residues by a cAMP-dependent protein kinase. Phosphorylation of P450c17 increases 17,20-lyase activity, while dephosphorylation virtually eliminates this activity. Hormonally regulated serine phosphorylation of human P450c17 suggests a possible mechanism for human adrenarche and may be a unifying etiologic link between the hyperandrogenism and insulin resistance that characterize the polycystic ovary syndrome. PMID- 7479853 TI - Identification of a DNA element determining synaptic expression of the mouse acetylcholine receptor delta-subunit gene. AB - mRNAs for acetylcholine receptor genes are highly concentrated in the endplate region of adult skeletal muscle largely as a result of a transcription restricted to the subneural nuclei. To identify the regulatory elements involved, we employed a DNA injection of a plasmid containing a fragment of the acetylcholine receptor delta-subunit gene promoter (positions -839 to +45) linked to the reporter gene lacZ with a nuclear localization signal. Injection of the wild-type construct into mouse leg muscles yielded preferential expression of the reporter gene in the synaptic region. Analysis of various mutant promoters resulted in the identification of a DNA element (positions -60 to -49), referred to as the N box, that plays a critical role in subneural expression. Disruption of this 12-bp element in the context of a mouse delta-subunit promoter from positions -839 to +45 gives widespread expression of the reporter gene throughout the entire muscle fiber, indicating that this element is a silencer that represses delta-subunit gene transcription in extrajunctional areas. On the other hand, this element inserted upstream of a heterologous basal promoter preferentially enhances expression in the endplate region. This element therefore regulates the restricted expression of the delta-subunit gene both as an enhancer at the endplate level and as a silencer in extrajunctional areas. Furthermore, gel-shift experiments with mouse muscle extracts reveal an activity that specifically binds the 6-bp sequence TTCCGG of this element, suggesting that a transcription factor(s) controls the expression of the delta-subunit gene via this element. PMID- 7479854 TI - Rho protein regulates tight junctions and perijunctional actin organization in polarized epithelia. AB - The rho family of GTP-binding proteins regulates actin filament organization. In unpolarized mammalian cells, rho proteins regulate the assembly of actin containing stress fibers at the cell-matrix interface. Polarized epithelial cells, in contrast, are tall and cylindrical with well developed intercellular tight junctions that permit them to behave as biologic barriers. We report that rho regulates filamentous actin organization preferentially in the apical pole of polarized intestinal epithelial cells and, in so doing, influences the organization and permeability of the associated apical tight junctions. Thus, barrier function, which is an essential characteristic of columnar epithelia, is regulated by rho. PMID- 7479855 TI - Point mutation of the autophosphorylation site or in the nuclear location signal causes protein kinase A RII beta regulatory subunit to lose its ability to revert transformed fibroblasts. AB - The RII beta regulatory subunit of cAMP-dependent protein kinase (PKA) contains an autophosphorylation site and a nuclear location signal, KKRK. We approached the structure-function analysis of RII beta by using site-directed mutagenesis. Ser114 (the autophosphorylation site) of human RII beta was replaced with Ala (RII beta-P) or Arg264 of KKRK was replaced with Met (RII beta-K). ras transformed NIH 3T3 (DT) cells were transfected with expression vectors for RII beta, RII beta-P, and RII beta-K, and the effects on PKA isozyme distribution and transformation properties were analyzed. DT cells contained PKA-I and PKA-II isozymes in a 1:2 ratio. Over-expression of wild-type or mutant RII beta resulted in an increase in PKA-II and the elimination of PKA-I. Only wild-type RII beta cells demonstrated inhibition of both anchorage-dependent and -independent growth and phenotypic change. The growth inhibitory effect of RII beta overexpression was not due to suppression of ras expression but was correlated with nuclear accumulation of RII beta. DT cells demonstrated growth inhibition and phenotypic change upon treatment with 8-Cl-cAMP. RII beta-P or RII beta-K cells failed to respond to 8-Cl-cAMP. These data suggest that autophosphorylation and nuclear location signal sequences are integral parts of the growth regulatory mechanism of RII beta. PMID- 7479856 TI - Modification of the substrate specificity of an acyl-acyl carrier protein thioesterase by protein engineering. AB - The plant acyl-acyl carrier protein (ACP) thioesterases (TEs) are of biochemical interest because of their roles in fatty acid synthesis and their utilities in the bioengineering of plant seed oils. When the FatB1 cDNA encoding a 12:0-ACP TE (Uc FatB1) from California bay, Umbellularia californica (Uc) was expressed in Escherichia coli and in developing oilseeds of the plants Arabidopsis thaliana and Brassica napus, large amounts of laurate (12:0) and small amounts of myristate (14:0) were accumulated. We have isolated a TE cDNA from camphor (Cinnamomum camphorum) (Cc) seeds that shares 92% amino acid identity with Uc FatB1. This TE, Cc FatB1, mainly hydrolyzes 14:0-ACP as shown by E. coli expression. We have investigated the roles of the N- and C-terminal regions in determining substrate specificity by constructing two chimeric enzymes, in which the N-terminal portion of one protein is fused to the C-terminal portion of the other. Our results show that the C-terminal two-thirds of the protein is critical for the specificity. By site-directed mutagenesis, we have replaced several amino acids in Uc FatB1 by using the Cc FatB1 sequence as a guide. A double mutant, which changes Met-197 to an Arg and Arg-199 to a His (M197R/R199H), turns Uc FatB1 into a 12:0/14:0 TE with equal preference for both substrates. Another mutation, T231K, by itself does not effect the specificity. However, when it is combined with the double mutant to generate a triple mutant (M197R/R199H/T231K), Uc FatB1 is converted to a 14:0-ACP TE. Expression of the double-mutant cDNA in E. coli K27, a strain deficient in fatty acid degradation, results in accumulation of similar amounts of 12:0 and 14:0. Meanwhile the E. coli expressing the triple-mutant cDNA produces predominantly 14:0 with very small amounts of 12:0. Kinetic studies indicate that both wild-type Uc FatB1 and the triple mutant have similar values of Km,app with respect to 14:0-ACP. Inhibitory studies also show that 12:0-ACP is a good competitive inhibitor with respect to 14:0-ACP in both the wild type and the triple mutant. These results imply that both 12:0- and 14:0-ACP can bind to the two proteins equally well, but in the case of the triple mutant, the hydrolysis of 12:0-ACP is severely impaired. The ability to modify TE specificity should allow the production of additional "designer oils" in genetically engineered plants. PMID- 7479857 TI - Target sequence recognition by the calmodulin superfamily: implications from light chain binding to the regulatory domain of scallop myosin. AB - Some of the rules for how members of the calmodulin (CaM) superfamily bind to target peptides are revealed by the crystal structure of the regulatory domain of scallop myosin. The structure shows that the IQ motif of the heavy chain in this invertebrate myosin imposes constraints on both the positioning and conformation of the individual lobes of the light chains. In contrast, analysis of the contact residues in the targets bound by Ca(2+)-CaM reveals how the structure of CaM accommodates a broader range of sequences consonant with this protein's functional diversity. PMID- 7479858 TI - Putative receptor binding sites on alphaviruses as visualized by cryoelectron microscopy. AB - The structures of Sindbis virus and Ross River virus complexed with Fab fragments from monoclonal antibodies have been determined from cryoelectron micrographs. Both antibodies chosen for this study bind to regions of the virions that have been implicated in cell-receptor recognition and recognize epitopes on the E2 glycoprotein. The two structures show that the Fab fragments bind to the outermost tip of the trimeric envelope spike protein. Hence, the same region of both the Sindbis virus and Ross River virus envelope spike is composed of E2 and is involved in recognition of the cellular receptor. PMID- 7479859 TI - Localization of vasopressin mRNA and immunoreactivity in pituicytes of pituitary stalk-transected rats after osmotic stimulation. AB - The presence of [arginine] vasopressin (AVP) mRNA and AVP immunoreactivity in pituicytes of the neural lobe (NL) of intact and pituitary stalk-transected rats, with and without osmotic stimulation, was examined. AVP mRNA was analyzed by Northern blotting, as well as by in situ hybridization in combination with immunocytochemistry using anti-glial fibrillary acidic protein (GFAP) as a marker for pituicytes. In intact rats, a poly(A) tail-truncated 0.62-kb AVP mRNA was detected in the NL and was found to increase 10-fold with 7 days of continuous salt loading. Morphological analysis of the NL of 7-day salt-loaded rats revealed the presence of AVP mRNA in a significant number of GFAP-positive pituicytes in the NL and in areas most probably containing nerve fibers. Eight days after pituitary stalk transection the NL AVP mRNA diminished in animals given water to drink, whereas in those given 2% saline for 18 h followed by 6 h of water, a treatment repeated on 6 successive days beginning 2 days after surgery, the 0.62 kb AVP mRNA was present. The AVP mRNA in the pituitary stalk-transected, salt loaded rats showed an exclusive cellular distribution in the NL, indicative of localization in pituicytes. Immunoelectron microscopy showed the presence of AVP immunoreactivity in a subpopulation of pituicytes 7 and 10 days after pituitary stalk transection in salt-loaded animals, when almost all AVP fibers had disappeared from the NL. These data show that a subset of pituicytes in the NL is activated to synthesize AVP mRNA and AVP in response to osmotic stimulation. PMID- 7479860 TI - An alternative pathway for signal flow from rod photoreceptors to ganglion cells in mammalian retina. AB - Rod signals in the mammalian retina are thought to reach ganglion cells over the circuit rod-->rod depolarizing bipolar cell-->AII amacrine cell-->cone bipolar cells-->ganglion cells. A possible alternative pathway involves gap junctions linking the rods and cones, the circuit being rod-->cone-->cone bipolar cells- >ganglion cells. It is not clear whether this second pathway indeed relays rod signals to ganglion cells. We studied signal flow in the isolated rabbit retina with a multielectrode array, which allows the activity of many identified ganglion cells to be observed simultaneously while the preparation is stimulated with light and/or exposed to drugs. When transmission between rods and rod depolarizing bipolar cells was blocked by the glutamate agonist 2-amino-4 phosphonobutyric acid (APB), rod input to all On-center and briskly responding Off-center ganglion cells was dramatically reduced as expected. Off responses persisted, however, in Off-center sluggish and On-Off direction-selective ganglion cells. Presumably these responses were generated by the alternative pathway involving rod-cone junctions. This APB-resistant pathway may carry the major rod input to Off-center sluggish and On-Off direction-selective ganglion cells. PMID- 7479861 TI - The angiotensin II type 2 (AT2) receptor antagonizes the growth effects of the AT1 receptor: gain-of-function study using gene transfer. AB - The type 1 angiotensin II (AT1) receptor is well characterized but the type 2 (AT2) receptor remains an enigma. We tested the hypothesis that the AT2 receptor can modulate the growth of vascular smooth muscle cells by transfecting an AT2 receptor expression vector into the balloon-injured rat carotid artery and observed that overexpression of the AT2 receptor attenuated neointimal formation. In cultured smooth muscle cells, AT2 receptor transfection reduced proliferation and inhibited mitogen-activated protein kinase activity. Furthermore, we demonstrated that the AT2 receptor mediated the developmentally regulated decrease in aortic DNA synthesis at the latter stages of gestation. These results suggest that the AT2 receptor exerts an antiproliferative effect, counteracting the growth action of AT1 receptor. PMID- 7479862 TI - Submillisecond events in protein folding. AB - The pathway of protein folding is now being analyzed at the resolution of individual residues by kinetic measurements on suitably engineered mutants. The kinetic methods generally employed for studying folding are typically limited to the time range of > or = 1 ms because the folding of denatured proteins is usually initiated by mixing them with buffers that favor folding, and the dead time of rapid mixing experiments is about a millisecond. We now show that the study of protein folding may be extended to the microsecond time region by using temperature-jump measurements on the cold-unfolded state of a suitable protein. We are able to detect early events in the folding of mutants of barstar, the polypeptide inhibitor of barnase. A preliminary characterization of the fast phase from spectroscopic and phi-value analysis indicates that it is a transition between two relatively solvent-exposed states with little consolidation of structure. PMID- 7479863 TI - Pituitary follicle-stimulating hormone (FSH) induces CREM gene expression in Sertoli cells: involvement in long-term desensitization of the FSH receptor. AB - Transcription factor CREM (cAMP-responsive element modulator) plays a pivotal role in the nuclear response to cAMP in neuroendocrine cells. We have previously shown that follicle-stimulating hormone (FSH) directs CREM expression in male germ cells. The physiological importance of FSH in Sertoli cell function prompted us to analyze its effect on CREM expression in these cells. We observed a dramatic and specific increase in the CREM isoform ICER (inducible cAMP early repressor) expression, with a peak 4 h after FSH treatment of primary Sertoli cells. Interestingly, induced levels of ICER protein persist for a considerably longer time. Induction of the repressor ICER accompanies early down-regulation of the FSH receptor transcript, which leads to long-term desensitization. Here we show that ICER represses FSH receptor expression by binding to a CRE-like sequence in the regulatory region of the gene. Our results confirm the crucial role played by CREM in hormonal control and suggest its role in the long-term desensitization phenomenon of peptide membrane receptors. PMID- 7479864 TI - Interaction between focal adhesion kinase and Crk-associated tyrosine kinase substrate p130Cas. AB - The focal adhesion kinase (FAK) has been implicated in integrin-mediated signaling events and in the mechanism of cell transformation by the v-Src and v Crk oncoproteins. To gain further insight into FAK signaling pathways, we used a two-hybrid screen to identify proteins that interact with mouse FAK. The screen identified two proteins that interact with FAK via their Src homology 3 (SH3) domains: a v-Crk-associated tyrosine kinase substrate (Cas), p130Cas, and a still uncharacterized protein, FIPSH3-2, which contains an SH3 domain closely related to that of p130Cas. These SH3 domains bind to the same proline-rich region of FAK (APPKPSR) encompassing residues 711-717. The mouse p130Cas amino acid sequence was deduced from cDNA clones, revealing an overall high degree of similarity to the recently reported rat sequence. Coimmunoprecipitation experiments confirmed that p130Cas and FAK are associated in mouse fibroblasts. The stable interaction between p130Cas and FAK emerges as a likely key element in integrin-mediated signal transduction and further represents a direct molecular link between the v Src and v-Crk oncoproteins. The Src family kinase Fyn, whose Src homology 2 (SH2) domain binds to the major FAK autophosphorylation site (tyrosine 397), was also identified in the two-hybrid screen. PMID- 7479865 TI - The activation domain of GAL4 protein mediates cooperative promoter binding with general transcription factors in vivo. AB - Most proteins that activate RNA polymerase II-mediated transcription in eukaryotic cells contain sequence-specific DNA-binding domains and "activation" regions. The latter bind general transcription factors and/or coactivators and are required for high-level transcription. Their function in vivo is unknown. Since several activation domains bind the TATA-binding protein (TBP), TBP associated factors, or other general factors in vitro, one role of the activation domain may be to facilitate promoter occupancy by supporting cooperative binding of the activator and general transcription factors. Using the GAL4 system of yeast, we have tested this model in vivo. It is demonstrated that the presence of a TATA box (the TBP binding site) facilitates binding of GAL4 protein to low- and moderate-affinity sites and that the activation domain modulates these effects. These results support the cooperative binding model for activation domain function in vivo. PMID- 7479866 TI - Mice lacking inducible nitric oxide synthase are not resistant to lipopolysaccharide-induced death. AB - Nitric oxide produced by cytokine-inducible nitric oxide synthase (iNOS) is thought to be important in the pathogenesis of septic shock. To further our understanding of the role of iNOS in normal biology and in a variety of inflammatory disorders, including septic shock, we have used gene targeting to generate a mouse strain that lacks iNOS. Mice lacking iNOS were indistinguishable from wild-type mice in appearance and histology. Upon treatment with lipopolysaccharide and interferon gamma, peritoneal macrophages from the mutant mice did not produce nitric oxide measured as nitrite in the culture medium. In addition, lysates of these cells did not contain iNOS protein by immunoblot analysis or iNOS enzyme activity. In a Northern analysis of total RNA, no iNOS transcript of the correct size was detected. No increases in serum nitrite plus nitrate levels were observed in homozygous mutant mice treated with a lethal dose of lipopolysaccharide, but the mutant mice exhibited no significant survival advantage over wild-type mice. These results show that lack of iNOS activity does not prevent mortality in this murine model for septic shock. PMID- 7479867 TI - Protective immune responses induced by secretion of a chimeric soluble protein from a recombinant Mycobacterium bovis bacillus Calmette-Guerin vector candidate vaccine for human immunodeficiency virus type 1 in small animals. AB - A recombinant Mycobacterium bovis bacillus Calmette-Guerin (BCG) vector-based vaccine that secretes the V3 principal neutralizing epitope of human immunodeficiency virus (HIV) could induce immune response to the epitope and prevent the viral infection. By using the Japanese consensus sequence of HIV-1, we successfully constructed chimeric protein secretion vectors by selecting an appropriate insertion site of a carrier protein and established the principal neutralizing determinant (PND)-peptide secretion system in BCG. The recombinant BCG (rBCG)-inoculated guinea pigs were initially screened by delayed-type hypersensitivity (DTH) skin reactions to the PND peptide, followed by passive transfer of the DTH by the systemic route. Further, immunization of mice with the rBCG resulted in induction of cytotoxic T lymphocytes. The guinea pig immune antisera showed elevated titers to the PND peptide and neutralized HIVMN, and administration of serum IgG from the vaccinated guinea pigs was effective in completely blocking the HIV infection in thymus/liver transplanted severe combined immunodeficiency (SCID)/hu or SCID/PBL mice. In addition, the immune serum IgG was shown to neutralize primary field isolates of HIV that match the neutralizing sequence motif by a peripheral blood mononuclear cell-based virus neutralization assay. The data support the idea that the antigen-secreting rBCG system can be used as a tool for development of HIV vaccines. PMID- 7479868 TI - Role of the C2A domain of synaptotagmin in transmitter release as determined by specific antibody injection into the squid giant synapse preterminal. AB - Squid synaptotagmin (Syt) cDNA, including its open reading frame, was cloned and polyclonal antibodies were obtained in rabbits immunized with glutathione S transferase (GST)-Syt-C2A. Binding assays indicated that the antibody, anti-Syt C2A, recognized squid Syt and inhibited the Ca(2+)-dependent phospholipid binding to the C2A domain. This antibody, when injected into the preterminal at the squid giant synapse, blocked transmitter release in a manner similar to that previously reported for the presynaptic injection of members of the inositol high polyphosphate series. The block was not accompanied by any change in the presynaptic action potential or the amplitude or voltage dependence of the presynaptic Ca2+ current. The postsynaptic potential was rather insensitive to repetitive presynaptic stimulation, indicating a direct effect of the antibody on the transmitter release system. Following block of transmitter release, confocal microscopical analysis of the preterminal junction injected with rhodamine conjugated anti-Syt-C2A demonstrated fluorescent spots at the inner surface of the presynaptic plasmalemma next to the active zones. Structural analysis of the same preparations demonstrated an accumulation of synaptic vesicles corresponding in size and distribution to the fluorescent spots demonstrated confocally. Together with the finding that such antibody prevents Ca2+ binding to a specific receptor in the C2A domain, these results indicate that Ca2+ triggers transmitter release by activating the C2A domain of Syt. We conclude that the C2A domain is directly related to the fusion of synaptic vesicles that results in transmitter release. PMID- 7479869 TI - Role of the C2B domain of synaptotagmin in vesicular release and recycling as determined by specific antibody injection into the squid giant synapse preterminal. AB - Synaptotagmin (Syt) is an inositol high-polyphosphate series [IHPS inositol 1,3,4,5-tetrakisphosphate (IP4), inositol 1,3,4,5,6-pentakisphosphate, and inositol 1,2,3,4,5,6-hexakisphosphate] binding synaptic vesicle protein. A polyclonal antibody against the C2B domain (anti-Syt-C2B), an IHPS binding site, was produced. The specificity of this antibody to the C2B domain was determined by comparing its ability to inhibit IP4 binding to the C2B domain with that to inhibit the Ca2+/phospholipid binding to the C2A domain. Injection of the anti Syt-C2B IgG into the squid giant presynapse did not block synaptic release. Coinjection of IP4 and anti-Syt-C2B IgG failed to block transmitter release, while IP4 itself was a powerful synpatic release blocker. Repetitive stimulation to presynaptic fiber injected with anti-Syt-C2B IgG demonstrated a rapid decline of the postsynaptic response amplitude probably due to its block of synaptic vesicle recycling. Electron microscopy of the anti-Syt-C2B-injected presynapse showed a 90% reduction of the numbers of synaptic vesicles. These results, taken together, indicate that the Syt molecule is central, in synaptic vesicle fusion by Ca2+ and its regulation by IHPS, as well as in the recycling of synaptic vesicles. PMID- 7479870 TI - Intraembryonic hematopoietic cell migration during vertebrate development. AB - Vertebrate hematopoietic stem cells are derived from vental mesoderm, which is postulated to migrate to both extra- and intraembryonic positions during gastrula and neurula stages. Extraembryonic migration has previously been documented, but the origin and migration of intraembryonic hematopoietic cells have not been visualized. The zebrafish and most other teleosts do not form yolk sac blood islands during early embryogenesis, but instead hematopoiesis occurs solely in a dorsal location known as the intermediate cell mass (IM) or Oellacher. In this report, we have isolated cDNAs encoding zebrafish homologs of the hematopoietic transcription factors GATA-1 and GATA-2 and have used these markers to determine that the IM is formed from mesodermal cells in a posterior-lateral position on the yolk syncytial layer of the gastrula yolk sac. Surprisingly, cells of the IM then migrate anteriorly through most of the body length prior to the onset of active circulation and exit onto the yolk sac. These findings support a hypothesis in which the hematopoietic program of vertebrates is established by variations in homologous migration pathways of extra- and intraembryonic progenitors. PMID- 7479871 TI - The hypocone as a key innovation in mammalian evolution. AB - The hypocone, a cusp added to the primitively triangular upper molar teeth of therian mammals, has evolved convergently > 20 times among mammals during the Cenozoic. Acquisition of the hypocone itself involves little phenotypic change, but subsequent diversification of groups possessing the hypocone may be greatly enhanced. Our analysis of the Cenozoic mammalian radiations, including the Recent fauna, shows that high species diversity of mammals with hypocones and association of the hypocone with herbivory strongly support recognition of the hypocone as a key innovation that has allowed invasion of, and diversification within, herbivorous adaptive zones. In contrast, mammals lacking hypocones show no marked increase in species diversity during the Cenozoic. PMID- 7479872 TI - Chimeric Na+/H+ exchangers: an epithelial membrane-bound N-terminal domain requires an epithelial cytoplasmic C-terminal domain for regulation by protein kinases. AB - All cloned members of the mammalian Na+/H+ exchanger gene family encode proteins that consist of two functionally distinct domains: a membrane-bound N terminus and a cytoplasmic C terminus, which are required for ion transport and regulation of transport, respectively. Despite their similarity in structure, three members of this family, designated NHE1, NHE2, and NHE3, exhibit different kinetic mechanisms in response to growth factors and protein kinases. For instance, growth factors stimulate NHE1 by a change in the affinity constant for intracellular H+, K'(Hi+), and regulate NHE2 and NHE3 by a change in Vmax. We have constructed chimeric Na+/H+ exchangers by exchanging the N and C termini among three cloned rabbit Na+/H+ exchangers (NHE1 to NHE3) to determine which domain is responsible for the above Vmax-vs.-K'(H(i)+) effect of the Na+/H+ isoforms. All of the chimeras had functional exchange activity and basal kinetic properties similar to those of wild-type exchangers. Studies with serum showed that the N terminus is responsible for the Vmax-vs.-K'(H(i)+) stimulation of the Na+/H+ exchanger isoforms. Moreover, phorbol 12-myristate 13-acetate and fibroblast growth factor altered Na+/H+ exchange only in chimeras that had an epithelial N-terminal domain matched with an epithelial C-terminal domain. Therefore, the protein kinase-induced regulation of Na+/H+ exchangers is mediated through a specific interaction between the N- and C-termini, whcih is restricted so that epithelial N- and epithelial N-and C-terminal portions of the exchangers are required for regulation. PMID- 7479873 TI - Glucose stimulation of insulin release in the absence of extracellular Ca2+ and in the absence of any increase in intracellular Ca2+ in rat pancreatic islets. AB - Insulin secretion has been studied in isolated rat pancreatic islets under stringent Ca(2+)-depleted, Ca(2+)-free conditions. Under these conditions, the effect of 16.7 mM glucose to stimulate insulin release was abolished. Forskolin, which activates adenylyl cyclase, also failed to stimulate release in the presence of either low or high glucose concentrations. A phorbol ester (phorbol 12-myristate 13-acetate; PMA) increased the release rate slightly and this was further increased by 16.7 mM glucose. Remarkably, in the presence of both forskolin and PMA, 16.7 mM glucose strongly augmented insulin release. The augmentation was concentration dependent and monophasic and had a temporal profile similar to the "second phase" of glucose-stimulated insulin release, which is seen under normal conditions when Ca2+ is present. Metabolism is required for the effect because mannoheptulose abolished the glucose response. Other nutrient secretagogues, alpha-ketoisocaproate, and the combination of leucine and glutamine augmented release under the same conditions. Norepinephrine, a physiological inhibitor of insulin secretion, totally blocked the stimulation of release by forskolin and PMA and the augmentation of release by glucose. Thus, under the stringent Ca(2+)-free conditions imposed, the stimulation of insulin release by forskolin and PMA, as well as the augmentation of release by glucose, is under normal physiological control. As no increase in intracellular [Ca2+] was observed, the results demonstrate that glucose can increase the rate of exocytosis and insulin release by pancreatic islets in a Ca(2+)-independent manner. This interesting pathway of stimulus-secretion coupling for glucose appears to exert its effect at a site beyond the usual elevation of intracellular [Ca2+] and is not due to an activation by glucose of protein kinase A or C. PMID- 7479874 TI - The rotated hypoblast of the chicken embryo does not initiate an ectopic axis in the epiblast. AB - In the amniotes, two unique layers of cells, the epiblast and the hypoblast, constitute the embryo at the blastula stage. All the tissues of the adult will derive from the epiblast, whereas hypoblast cells will form extraembryonic yolk sac endoderm. During gastrulation, the endoderm and the mesoderm of the embryo arise from the primitive streak, which is an epiblast structure through which cells enter the interior. Previous investigations by others have led to the conclusion that the avian hypoblast, when rotated with regard to the epiblast, has inductive properties that can change the fate of competent cells in the epiblast to form an ectopic embryonic axis. Thus, it has been suggested that the hypoblast normally induces the epiblast to form a primitive streak at a specific locus. In the work reported here, an attempt was made to reexamine the issue of induction. In contrast to previous reports, it was found that the rotated hypoblast of the chicken embryo does not initiate formation of an ectopic axis in the epiblast. The embryonic axis always initiates and develops according to the basic polarity of the epiblast layer. These results provoke a reinterpretation of the issues of mesoderm induction and primitive streak initiation in the avian embryo. PMID- 7479875 TI - Amyloid beta peptide potentiates cytokine secretion by interleukin-1 beta activated human astrocytoma cells. AB - Neurodegenerative processes in Alzheimer disease (AD) are thought to be driven in part by the deposition of amyloid beta (A beta), a 39- to 43-amino acid peptide product resulting from an alternative cleavage of amyloid precursor protein. Recent descriptions of in vitro neurotoxic effects of A beta support this hypothesis and suggest toxicity might be mediated by A beta-induced neuronal calcium disregulation. In addition, it has been reported that "aging" A beta results in increased toxic potency due to peptide aggregation and formation of a beta-sheet secondary structure. In addition, A beta might also promote neuropathology indirectly by activating immune/inflammatory pathways in affected areas of the brain (e.g., cortex and hippocampus). Here we report that A beta can modulate cytokine secretion [interleukins 6 and 8 (IL-6 and IL-8)] from human astrocytoma cells (U-373 MG). Freshly prepared and aged A beta modestly stimulated IL-6 and IL-8 secretion from U-373 MG cells. However, in the presence of interleukin-1 beta (IL-1 beta), aged, but not fresh, A beta markedly potentiated (3- to 8-fold) cytokine release. In contrast, aged A beta did not potentiate substance P (NK-1)- or histamine (H1)-stimulated cytokine production. Further studies showed that IL-1 beta-induced cytokine release was potentiated by A beta-(25-35), while A beta-(1-16) was inactive. Calcium disregulation may be responsible for the effects of A beta on cytokine production, since the calcium ionophore A23187 similarly potentiated IL-1 beta-induced cytokine secretion and EGTA treatment blocked either A beta or A23187 activity. Thus, chronic neurodegeneration in AD-affected brain regions may be mediated in part by the ability of A beta to exacerbate inflammatory pathways in a conformation-dependent manner. PMID- 7479876 TI - Changing nucleosome positions through modification of the DNA rotational information. AB - The effects of the rotational information of DNA in determining the in vitro localization of nucleosomal core particles (ncps) have been studied in the Saccharomyces cerevisiae 5S rRNA repeat gene. We have altered the distribution of the phased series of flexibility signals present on this DNA by inserting a 25-bp tract, and we have analyzed the effects of this mutation on the distribution and on the frequencies of ncps, as compared with the wild type and a reference 21-bp insertion mutant. The variation of the standard free energy of nucleosome reconstitution was determined. The results show that the DNA rotational information is a major determinant of ncps positioning, define how many rotationally phased signals are required for the formation of a stable particle, and teach how to modify their distribution through the alteration of the rotational signals. PMID- 7479877 TI - Shared functions in vivo of a glycosyl-phosphatidylinositol-linked aspartyl protease, Mkc7, and the proprotein processing protease Kex2 in yeast. AB - The MKC7 gene was isolated as a multicopy suppressor of the cold-sensitive growth phenotype of a yeast kex2 mutant, which lacks the protease that cleaves pro-alpha factor and other secretory proproteins at pairs of basic residues in a late Golgi compartment in yeast. MKC7 encodes an aspartyl protease most closely related to product of the YAP3 gene, a previously isolated multicopy suppressor of the pro alpha-factor processing defect of a kex2 null. Multicopy MKC7 suppressed the alpha-specific mating defect of a kex2 null as well as multicopy YAP3 did, but multicopy YAP3 was a relatively weak suppressor of kex2 cold sensitivity. Overexpression of MKC7 resulted in production of a membrane-associated proteolytic activity that cleaved an internally quenched fluorogenic peptide substrate on the carboxyl side of a Lys-Arg site. Treatment with phosphatidylinositol-specific phospholipase C shifted Mkc7 activity from the detergent to the aqueous phase in a Triton X-114 phase separation, indicating that membrane attachment of Mkc7 is mediated by a glycosyl-phosphatidylinositol anchor. Although disruption of MKC7 or YAP3 alone resulted in no observable phenotype, mkc7 yap3 double disruptants exhibited impaired growth at 37 degrees C. Disruption of MKC7 and YAP3 in a kex2 null mutant resulted in profound temperature sensitivity and more generalized cold sensitivity. The synergism of mkc7, yap3, and kex2 null mutations argues that Mkc7 and Yap3 are authentic processing enzymes whose functions overlap those of Kex2 in vivo. PMID- 7479878 TI - Emergence of the ZNF91 Kruppel-associated box-containing zinc finger gene family in the last common ancestor of anthropoidea. AB - The ZNF91 gene family, a subset of the Kruppel-associated box (KRAB)-containing group of zinc finger genes, comprises more than 40 loci; most reside on human chromosome 19p12-p13.1. We have examined the emergence and evolutionary conservation of the ZNF91 family. ZNF91 family members were detected in all species of great apes, gibbons, Old World monkeys, and New World monkeys examined but were not found in prosimians or rodents. In each species containing the ZNF91 family, the genes were clustered at one major site, on the chromosome(s) syntenic to human chromosome 19. To identify a putative "founder" gene, > 20 murine KRAB containing zinc finger protein (ZFP) cDNAs were randomly cloned, but none showed sequence similarity to the ZNF91 genes. These observations suggest that the ZNF91 gene cluster is a derived character specific to Anthropoidea, resulting from a duplication and amplification event some 55 million years ago in the common ancestor of simians. Although the ZNF91 gene cluster is present in all simian species, the sequences of the human ZNF91 gene that confer DNA-binding specificity were conserved only in great apes, suggesting that there is not a high selective pressure to maintain the DNA targets of these proteins during evolution. PMID- 7479880 TI - Crystal structure of a DNA decamer showing a novel pseudo four-way helix-helix junction. AB - The crystal structure of the decanucleotide d(CGCAATTGCG)2 has been solved by a combination of molecular replacement and heavy-atom procedures and has been refined to an R factor of 20.2% at 2.7 A. It is not a fully base-paired duplex but has a central core of eight Watson-Crick base pairs flanked by unpaired terminal guanosines and cytosines. These participate in hydrogen-bonding arrangements with adjacent decamer duplexes in the crystal lattice. The unpaired guanosines are bound in the G+C regions of duplex minor grooves. The cytosines have relatively high mobility, even though they are constrained to be in one region where they are involved in base-paired triplets with G.C base pairs. The 5'-AATT sequence in the duplex region has a narrow minor groove, providing further confirmation of the sequence-dependent nature of groove width. PMID- 7479879 TI - Crystal structure of Pseudomonas aeruginosa catabolic ornithine transcarbamoylase at 3.0-A resolution: a different oligomeric organization in the transcarbamoylase family. AB - The crystal structure of the Glu-105-->Gly mutant of catabolic ornithine transcarbamoylase (OTCase; carbamoyl phosphate + L-ornithine = orthophosphate + L citrulline, EC 2.1.3.3) from Pseudomonas aeruginosa has been determined at 3.0-A resolution. This mutant is blocked in the active R (relaxed) state. The structure was solved by the molecular replacement method, starting from a crude molecular model built from a trimer of the catalytic subunit of another transcarbamoylase, the extensively studied aspartate transcarbamoylase (ATCase) from Escherichia coli. This model was used to generate initial low-resolution phases at 8-A resolution, which were extended to 3-A by noncrystallographic symmetry averaging. Four phase extensions were required to obtain an electron density map of very high quality from which the final model was built. The structure, including 4020 residues, has been refined to 3-A, and the current crystallographic R value is 0.216. No solvent molecules have been added to the model. The catabolic OTCase is a dodecamer composed of four trimers organized in a tetrahedral manner. Each monomer is composed of two domains. The carbamoyl phosphate binding domain shows a strong structural homology with the equivalent ATCase part. In contrast, the other domain, mainly implicated in the binding of the second substrate (ornithine for OTCase and aspartate for ATCase) is poorly conserved. The quaternary structures of these two allosteric transcarbamoylases are quite divergent: the E. coli ATCase has pseudo-32 point-group symmetry, with six catalytic and six regulatory chains; the catabolic OTCase has 23 point-group symmetry and only catalytic chains. However, both enzymes display homotropic and heterotropic cooperativity. PMID- 7479881 TI - Interleukin 2 activates STAT5 transcription factor (mammary gland factor) and specific gene expression in T lymphocytes. AB - Although prolactin and interleukin 2 (IL-2) can elicit distinct physiological responses, we have found that their signal pathways share a common signal transducer and activator of transcription, STAT5. STAT5 was originally identified as a mammary gland factor induced by prolactin in lactating breast cells. Here we demonstrate that STAT5 is activated after IL-2 stimulation of two responsive lymphocyte cell lines, Nb2 and YT. Activation of STAT5 is measured both by IL-2 induced tyrosine phosphorylation and by IL-2-induced DNA binding. The STAT5 DNA recognition site is the same as the interferon gamma-activated site (GAS) in the interferon regulatory factor 1 gene. We demonstrate that the GAS element is necessary and sufficient for transcriptional induction by both IL-2 and prolactin in T lymphocytes. These results indicate that the role of STAT5 in the regulation of gene expression is not restricted to mammary cells or to prolactin, but is an integral part of the signal pathway of a critical immunomodulatory cytokine, IL 2. PMID- 7479882 TI - Cytoplasmic free-Ca2+ level rises with repellents and falls with attractants in Escherichia coli chemotaxis. AB - Cytoplasmic free-Ca2+ levels in Escherichia coli were measured by use of the fluorescent Ca(2+)-indicator dye fura-2. Chemotactically wild-type E. coli regulated cytoplasmic free Ca2+ at approximately 100 nM when no stimuli were encountered, but changes in bacterial behavior correlated with changes in cytoplasmic free-Ca2+ concentration. For chemotactically wild-type E. coli, addition of a repellent resulted in cells tumbling and a transient increase in cytoplasmic free-Ca2+ levels. Conversely, addition of an attractant to wild-type cells caused running and produced a transient decrease in cytoplasmic free-Ca2+ levels. Studies with mutant strains showed that the chemoreceptors were required for the observed changes in cytoplasmic free-Ca2+ levels in response to chemical stimuli. PMID- 7479884 TI - exl protein specifically binds BLE1, a bicoid mRNA localization element, and is required for one phase of its activity. AB - Localization of mRNAs, a crucial step in the early development of some animals, has been shown to be directed by cis-acting elements that presumably interact with localization factors. Here we identify a protein, exl, that binds to BLE1, an RNA localization element from the Drosophila bicoid mRNA. Using mutations in BLE1, we demonstrate a correlation between in vitro exl binding and one phase of in vivo localization directed by BLE1, implicating exl in that localization event. Furthermore, the same phase of localization is disrupted in exuperantia mutants, suggesting that exl and exuperantia proteins interact. Identification of a protein that binds specifically to an mRNA localization element and acts in mRNA localization opens the way for a biochemical analysis of this process. PMID- 7479883 TI - Distribution of parthenogenetic cells in the mouse brain and their influence on brain development and behavior. AB - A systematic analysis of parthenogenetic (PG) cell fate within the central nervous system (CNS) was made throughout fetal development and neonatal and adult life. Chimeras were made between PG embryos carrying a ubiquitously expressed lacZ transgene and normal fertilized embryos. After detailed histological analysis, we find that the developmental potential of PG cells is spatially restricted to certain parts of the brain. PG cells are prevalent in telencephalic structures and are largely excluded from diencephalic structures, especially the hypothalamus. These spatial restrictions are established early in development. Behavioral studies with chimeras identified an increase in male aggression when the proportion of PG cells in the brain was high. These studies demonstrate that imprinted genes play key roles in development of the CNS and may be involved in behavior. PMID- 7479885 TI - Gene for the catalytic subunit of mouse DNA-dependent protein kinase maps to the scid locus. AB - The gene encoding the catalytic subunit of DNA-dependent protein kinase (DNA PKcs) has been proposed recently as a candidate gene for the mouse severe combined immune deficiency (scid) locus. We have used a partial cDNA clone for human DNA-PKcs to map the mouse homologue using a large interspecific backcross panel. We found that the mouse gene for DNA-PKcs does not recombine with scid, consistent with the hypothesis that scid is a mutation in the mouse gene for DNA PKcs. PMID- 7479886 TI - Gene conversions and unequal crossovers between CYP21 (steroid 21-hydroxylase gene) and CYP21P involve different mechanisms. AB - Most cases of congenital adrenal hyperplasia, the inherited inability to synthesize cortisol, are caused by mutations in the steroid 21-hydroxylase gene (CYP21). Steroid 21-hydroxylase deficiency is unusual among genetic diseases in that approximately 95% of the mutant alleles have apparently been generated by recombination between a normally active gene (CYP21) and a linked pseudogene (CYP21P). Approximately 20% of mutant alleles carry DNA deletions of 30 kb that have presumably been generated by unequal meiotic crossing-over, whereas 75% carry one or more mutations in CYP21 that are normally found in the CYP21P pseudogene. These latter mutations are termed "gene conversions," although the mechanism by which they are generated is not well understood. To assess the frequency at which these different recombination events occur, we have used PCR to detect de novo deletions and gene conversions in matched sperm and peripheral blood leukocyte DNA samples from normal individuals. Deletions with breakpoints in a 100-bp region in intron 2 and exon 3 were detected in sperm DNA samples with frequencies of approximately 1 in 10(5)-10(6) genomes but were never detected in the matching leukocyte DNA. Gene conversions in the same region occur in approximately 1 in 10(3)-10(5) genomes in both sperm and leukocyte DNA. These data suggest that whereas deletions occur exclusively in meiosis, gene conversions occur during both meiosis and mitosis, or perhaps only during mitosis. Thus, gene conversions must occur by a mechanism distinct from unequal crossing-over. PMID- 7479887 TI - Two subsites in the binding domain of the acetylcholine receptor: an aromatic subsite and a proline subsite. AB - The ligand binding site of the nicotinic acetylcholine receptor (AcChoR) is localized in the alpha-subunit within a domain containing the tandem Cys-192 and 193. By analyzing the binding-site region of AcChoR from animal species that are resistant to alpha-neurotoxins, we have previously shown that four residues in this region, at positions 187, 189, 194, and 197, differ between animals sensitive (e.g., mouse) and resistant (e.g., mongoose and snake) to alpha bungarotoxin (alpha-BTX). In the present study, we performed site-directed mutagenesis on a fragment of the mongoose AcChoR alpha-subunit (residues 122-205) and exchanged residues 187, 189, 194, and 197, either alone or in combination, with those present in the mouse alpha-subunit sequence. Only the mongoose fragment in which all four residues were mutated to the mouse ones exhibited alpha-BTX binding similar to that of the mouse fragment. The mongoose double mutation in which Leu-194 and His-197 were replaced with proline residues, which are present at these positions in the mouse AcChoR and in all other toxin binders, bound alpha-BTX to approximately 60% of the level of binding exhibited by the mouse fragment. In addition, replacement of either Pro-194 or -197 in the mouse fragment with serine and histidine, respectively, markedly decreased alpha BTX binding. All other mutations resulted in no or just a small increase in alpha BTX binding. These results have led us to propose two subsites in the binding domain for alpha-BTX: the proline subsite, which includes Pro-194 and -197 and is critical for alpha-BTX binding, and the aromatic subsite, which includes amino acid residues 187 and 189 and determines the extent of alpha-BTX binding. PMID- 7479888 TI - Pregnenolone sulfate enhances post-training memory processes when injected in very low doses into limbic system structures: the amygdala is by far the most sensitive. AB - Immediate post-training, stereotactically guided, intraparenchymal administration of pregnenolone sulfate (PS) into the amygdala, septum, mammillary bodies, or caudate nucleus and of PS, dehydroepiandrosterone sulfate, and corticosterone into the hippocampus was performed in mice that had been weakly trained in a foot shock active avoidance paradigm. Intrahippocampal injection of PS resulted in memory enhancement (ME) at a lower dose than was found with dehydroepiandrosterone sulfate and corticosterone. Intraamygdally administered PS was approximately 10(4) times more potent on a molar basis in producing ME than when PS was injected into the hippocampus and approximately 10(5) times more potent than when injected into the septum or mammillary bodies. ME did not occur on injection of PS into the caudate nucleus over the range of doses tested in the other brain structures. The finding that fewer than 150 molecules of PS significantly enhanced post-training memory processes when injected into the amygdala establishes PS as the most potent memory enhancer yet reported and the amygdala as the most sensitive brain region for ME by any substance yet tested. PMID- 7479890 TI - Asbestos and mesothelioma: genetic lessons from a tragedy. PMID- 7479891 TI - Ahead of schedule and under budget: the Genome Project passes its fifth birthday. PMID- 7479889 TI - Self-recognition in primates: phylogeny and the salience of species-typical features. AB - Self-recognition has been explored in nonlinguistic organisms by recording whether individuals touch a dye-marked area on visually inaccessible parts of their face while looking in a mirror or inspect parts of their body while using the mirror's reflection. Only chimpanzees, gorillas, orangutans, and humans over the age of approximately 2 years consistently evidence self-directed mirror guided behavior without experimenter training. To evaluate the inferred phylogenetic gap between hominoids and other animals, a modified dye-mark test was conducted with cotton-top tamarins (Saguinus oedipus), a New World monkey species. The white hair on the tamarins' head was color-dyed, thereby significantly altering a visually distinctive species-typical feature. Only individuals with dyed hair and prior mirror exposure touched their head while looking in the mirror. They looked longer in the mirror than controls, and some individuals used the mirror to observe visually inaccessible body parts. Prior failures to pass the mirror test may have been due to methodological problems, rather than to phylogenetic differences in the capacity for self-recognition. Specifically, an individual's sensitivity to experimentally modified parts of its body may depend crucially on the relative saliency of the modified part (e.g., face versus hair). Moreover, and in contrast to previous claims, we suggest that the mirror test may not be sufficient for assessing the concept of self or mental state attribution in nonlinguistic organisms. PMID- 7479893 TI - The genome of Arabidopsis thaliana. AB - Arabidopsis thaliana is a small flowering plant that is a member of the family cruciferae. It has many characteristics--diploid genetics, rapid growth cycle, relatively low repetitive DNA content, and small genome size--that recommend it as the model for a plant genome project. The current status of the genetic and physical maps, as well as efforts to sequence the genome, are presented. Examples are given of genes isolated by using map-based cloning. The importance of the Arabidopsis project for plant biology in general is discussed. PMID- 7479892 TI - Gene disruptions using P transposable elements: an integral component of the Drosophila genome project. AB - Biologists require genetic as well as molecular tools to decipher genomic information and ultimately to understand gene function. The Berkeley Drosophila Genome Project is addressing these needs with a massive gene disruption project that uses individual, genetically engineered P transposable elements to target open reading frames throughout the Drosophila genome. DNA flanking the insertions is sequenced, thereby placing an extensive series of genetic markers on the physical genomic map and associating insertions with specific open reading frames and genes. Insertions from the collection now lie within or near most Drosophila genes, greatly reducing the time required to identify new mutations and analyze gene functions. Information revealed from these studies about P element site specificity is being used to target the remaining open reading frames. PMID- 7479894 TI - The genome of Caenorhabditis elegans. AB - The physical map of the 100-Mb Caenorhabditis elegans genome consists of 17,500 cosmids and 3500 yeast artificial chromosomes (YACs). A total of 22.5 Mb has been sequenced, with the remainder expected by 1998. A further 15.5 Mb of unfinished sequence is freely available online: because the areas sequenced so far are relatively gene rich, about half the 13,000 genes can now be scanned. More than a quarter of the genes are represented by expressed sequence tags (ESTs). All information pertaining to the genome is publicly available in the ACeDB data base. PMID- 7479895 TI - How is the Human Genome Project doing, and what have we learned so far? AB - In this paper, we describe the accomplishments of the initial phase of the Human Genome Project, with particular attention to the progress made toward achieving the defined goals for constructing genetic and physical maps of the human genome and determining the sequence of human DNA, identifying the complete set of human genes, and analyzing the need for adequate policies for using the information about human genetics in ways that maximize the benefits for individuals and society. PMID- 7479896 TI - Mapping the mouse genome: current status and future prospects. AB - The mouse is the best model system for the study of mammalian genetics and physiology. Because of the feasibility and importance of studying genetic crosses, the mouse genetic map has received tremendous attention in recent years. It currently contains over 14,000 genetically mapped markers, including 700 mutant loci, 3500 genes, and 6500 simple sequence length polymorphisms (SSLPs). The mutant loci and genes allow insights and correlations concerning physiology and development. The SSLPs provide highly polymorphic anchor points that allow inheritance to be traced in any cross and provide a scaffold for assembling physical maps. Adequate physical mapping resources--notably large-insert yeast artificial chromosome (YAC) libraries--are available to support positional cloning projects based on the genetic map, but a comprehensive physical map is still a few years away. Large-scale sequencing efforts have not yet begun in mouse, but comparative sequence analysis between mouse and human is likely to provide tremendous information about gene structure and regulation. PMID- 7479898 TI - Catalytic editing properties of DNA polymerases. AB - Enzymatic incorporation of 2',3'-dideoxynucleotides into DNA results in chain termination. We report that 3'-esterified 2'-deoxynucleoside 5'-triphosphates (dNTPs) are false chain-terminator substrates since DNA polymerases, including human immunodeficiency virus reverse transcriptase, can incorporate them into DNA and, subsequently, use this new 3' end to insert the next correctly paired dNTP. Likewise, a DNA substrate with a primer chemically esterified at the 3' position can be extended efficiently upon incubation with dNTPs and T7 DNA polymerase lacking 3'-to-5' exonuclease activity. This enzyme is also able to use dTTP bearing reporter groups in the 3' position conjugated through amide or thiourea bonds and cleave them to restore a DNA chain terminated by an amino group at the 3' end. Hence, a number of DNA polymerases exhibit wide catalytic versatility at the 3' end of the nascent DNA strand. As part of the polymerization mechanism, these capabilities extend the number of enzymatic activities associated with these enzymes and also the study of interactions between DNA polymerases and nucleotide analogues. PMID- 7479897 TI - High frequency of inactivating mutations in the neurofibromatosis type 2 gene (NF2) in primary malignant mesotheliomas. AB - Malignant mesotheliomas (MMs) are aggressive tumors that develop most frequently in the pleura of patients exposed to asbestos. In contrast to many other cancers, relatively few molecular alterations have been described in MMs. The most frequent numerical cytogenetic abnormality in MMs is loss of chromosome 22. The neurofibromatosis type 2 gene (NF2) is a tumor suppressor gene assigned to chromosome 22q which plays an important role in the development of familial and spontaneous tumors of neuroectodermal origin. Although MMs have a different histogenic derivation, the frequent abnormalities of chromosome 22 warranted an investigation of the NF2 gene in these tumors. Both cDNAs from 15 MM cell lines and genomic DNAs from 7 matched primary tumors were analyzed for mutations within the NF2 coding region. NF2 mutations predicting either interstitial in-frame deletions or truncation of the NF2-encoded protein (merlin) were detected in eight cell lines (53%), six of which were confirmed in primary tumor DNAs. In two samples that showed NF2 gene transcript alterations, no genomic DNA mutations were detected, suggesting that aberrant splicing may constitute an additional mechanism for merlin inactivation. These findings implicate NF2 in the oncogenesis of primary MMs and provide evidence that this gene can be involved in the development of tumors other than nervous system neoplasms characteristic of the NF2 disorder. In addition, unlike NF2-related tumors, MM derives from the mesoderm; malignancies of this origin have not previously been associated with frequent alterations of the NF2 gene. PMID- 7479899 TI - Yeast global transcriptional regulators Sin4 and Rgr1 are components of mediator complex/RNA polymerase II holoenzyme. AB - Sin4 and Rgr1 proteins, previously shown by genetic studies to play both positive and negative roles in the transcriptional regulation of many genes, are identified here as components of mediator and RNA polymerase II holoenzyme complexes. Results with Sin4 deletion and Rgr1 truncation strains indicate the association of these proteins in a subcomplex comprising Sin4, Rgr1, Gal11, and a 50-kDa polypeptide. Taken together with the previous genetic evidence, our findings point to a role of the mediator in repression as well as in transcriptional activation. PMID- 7479900 TI - Optimization of rates of protein folding: the nucleation-condensation mechanism and its implications. AB - Small, single-module proteins that fold in a single cooperative step may be paradigms for understanding early events in protein-folding pathways generally. Recent experimental studies of the 64-residue chymotrypsin inhibitor 2 (CI2) support a nucleation mechanism for folding, as do some computer stimulations. CI2 has a nucleation site that develops only in the transition state for folding. The nucleus is composed of a set of adjacent residues (an alpha-helix), stabilized by long-range interactions that are formed as the rest of the protein collapses around it. A simple analysis of the optimization of the rate of protein folding predicts that rates are highest when the denatured state has little residual structure under physiological conditions and no intermediates accumulate. This implies that any potential nucleation site that is composed mainly of adjacent residues should be just weakly populated in the denatured state and become structured only in a high-energy intermediate or transition state when it is stabilized by interactions elsewhere in the protein. Hierarchical mechanisms of folding in which stable elements of structure accrete are unfavorable. The nucleation-condensation mechanism of CI2 fulfills the criteria for fast folding. On the other hand, stable intermediates do form in the folding of more complex proteins, and this may be an unavoidable consequence of increasing size and nucleation at more than one site. PMID- 7479902 TI - Mutagenesis in the C-terminal region of human interleukin 5 reveals a central patch for receptor alpha chain recognition. AB - Cassette mutagenesis was used to identify side chains in human interleukin 5 (hIL 5) that mediate binding to hIL-5 receptor alpha chain (hIL-5R alpha). A series of single alanine substitutions was introduced into a stretch of residues in the C terminal region, including helix D, which previously had been implicated in receptor alpha chain recognition and which is aligned on the IL-5 surface so as to allow the topography of receptor binding residues to be examined. hIL-5 and single site mutants were expressed in COS cells, their interactions with hIL-5R alpha were measured by a sandwich surface plasmon resonance biosensor method, and their biological activities were measured by an IL-5-dependent cell proliferation assay. A pattern of mutagenesis effects was observed, with greatest impact near the interface between the two four-helix bundles of IL-5, in particular at residues Glu-110 and Trp-111, and least at the distal ends of the D helices. This pattern suggests the possibility that residues near the interface of the two four helix bundles in hIL-5 comprise a central patch or hot spot, which constitutes an energetically important alpha chain recognition site. This hypothesis suggests a structural explanation for the 1:1 stoichiometry observed for the complex of hIL 5 with hIL-5R alpha. PMID- 7479901 TI - Role of insulin-like growth factors and myogenin in the altered program of proliferation and differentiation in the NFB4 mutant muscle cell line. AB - In the present study we used the mutant muscle cell line NFB4 to study the balance between proliferation and myogenic differentiation. We show that removal of serum, which induced the parental C2C12 cells to withdraw from the cell cycle and differentiate, had little effect on NFB4 cells. Gene products characteristic of the proliferation state, such as c-Jun, continued to accumulate in the mutant cells in low serum, whereas those involved in differentiation, like myogenin, insulin-like growth factor II (IGF-II), and IGF-binding protein 5 (IGFBP-5) were undetectable. Moreover, NFB4 cells displayed a unique pattern of tyrosine phosphorylated proteins, especially in low serum, suggesting that the signal transduction pathway(s) that controls differentiation is not properly regulated in these cells. Treatment of NFB4 cells with exogenous IGF-I or IGF-II at concentrations shown to promote myogenic differentiation in wild-type cells resulted in activation of myogenin but not MyoD gene expression, secretion of IG FBP-5, changes in tyrosine phosphorylation, and enhanced myogenic differentiation. Similarly, transfection of myogenin expression constructs also enhanced differentiation and resulted in activation of IGF-II expression, showing that myogenin and IGF-II cross-activate each other's expression. However, in both cases, the expression of Jun mRNA remained elevated, suggesting that IGFs and myogenin cannot overcome all aspects of the block to differentiation in NFB4 cells. PMID- 7479903 TI - Analysis of the relation between the sequence and secondary and three-dimensional structures of immunoglobulin molecules. AB - Methods of structural and statistical analysis of the relation between the sequence and secondary and three-dimensional structures are developed. About 5000 secondary structures of immunoglobulin molecules from the Kabat data base were predicted. Two statistical analyses of amino acids reveal 47 universal positions in strands and loops. Eight universally conservative positions out of the 47 are singled out because they contain the same amino acid in > 90% of all chains. The remaining 39 positions, which we term universally alternative positions, were divided into five groups: hydrophobic, charged and polar, aromatic, hydrophilic, and Gly-Ala, corresponding to the residues that occupied them in almost all chains. The analysis of residue-residue contacts shows that the 47 universal positions can be distinguished by the number and types of contacts. The calculations of contact maps in the 29 antibody structures revealed that residues in 24 of these 47 positions have contacts only with residues of antiparallel beta strands in the same beta-sheet and residues in the remaining 23 positions always have far-away contacts with residues from other beta-sheets as well. In addition, residues in 6 of the 47 universal positions are also involved in interactions with residues of the other variable or constant domains. PMID- 7479905 TI - Genomic structure of human microtubule-associated protein 2 (MAP-2) and characterization of additional MAP-2 isoforms. AB - We have determined that the gene for human microtubule-associated protein 2 (MAP 2) spans 19 exons, including 6 exons identified in this study, 1-4, 8, and 13; all six of these exons are transcribed. The alternative splicing of coding exons generates a greater diversity of MAP-2 transcripts and isoforms. The first three exons encode alternate 5' untranslated regions that can be spliced to additional untranslated sequences contained in exons 4 and 5. Exons 8 and 13 are transcribed in human fetal spinal cord, adult brain, MSN cells, and rat brain, and each exon maintains an open reading frame with both high and low molecular weight MAP-2 isoforms. Antibodies generated to synthetic peptides of exons 8 and 13 demonstrate that these exons are translated and MAP-2 isoforms containing these exons are generated. PMID- 7479904 TI - Mutant forms of growth factor-binding protein-2 reverse BCR-ABL-induced transformation. AB - Growth factor-binding protein 2 (Grb2) is an adaptor protein that links tyrosine kinases to Ras. BCR-ABL is a tyrosine kinase oncoprotein that is implicated in the pathogenesis of Philadelphia chromosome (Ph1)-positive leukemias. Grb2 forms a complex with BCR-ABL and the nucleotide exchange factor Sos that leads to the activation of the Ras protooncogene. In this report we demonstrate that Grb2 mutant proteins lacking amino- or carboxyl-terminal src homology SH3 domains suppress BCR-ABL-induced Ras activation and reverse the oncogenic phenotype. The Grb2 SH3-deletion mutant proteins bind to BCR-ABL and do not impair tyrosine kinase activity. Expression of the Grb2 SH3-deletion mutant proteins in BCR-ABL transformed Rat-1 fibroblasts and in the human Ph1-positive leukemic cell line K562 inhibits their ability to grow as foci in soft agar and form tumors in nude mice. Furthermore, expression of the Grb2 SH3-deletion mutants in K562 cells induced their differentiation. Because Ras plays an important role in signaling by receptor and nonreceptor tyrosine kinases, the use of interfering mutant Grb2 proteins may be applied to block the proliferation of other cancers that depend in part on activated tyrosine kinases for growth. PMID- 7479906 TI - Fixing human factor IX (fIX): correction of a cryptic RNA splice enables the production of biologically active fIX in the mammary gland of transgenic mice. AB - Transgenic mice and sheep secrete only low levels of human factor IX in their milk because of an aberrant splicing of the transgene RNA in the mammary gland. Removal of the cryptic 3' splice site prevents this splicing and leads to the production of relatively high levels of factor IX. The purified protein is fully active showing that the mammary gland is capable of the efficient post translational modification of this protein and that transgenic animals are a suitable means of its production. PMID- 7479907 TI - The vertebrate alcohol dehydrogenase system: variable class II type form elucidates separate stages of enzymogenesis. AB - A mixed-class alcohol dehydrogenase has been characterized from avian liver. Its functional properties resemble the classical class I type enzyme in livers of humans and animals by exhibiting low Km and kcat values with alcohols (Km = 0.7 mM with ethanol) and low Ki values with 4-methylpyrazole (4 microM). These values are markedly different from corresponding parameters of class II and III enzymes. In contrast, the primary structure of this avian liver alcohol dehydrogenase reveals an overall relationship closer to class II and to some extent class III (69 and 65% residue identities, respectively) than to class I or the other classes of the human alcohol dehydrogenases (52-61%), the presence of an insertion (four positions in a segment close to position 120) as in class II but in no other class of the human enzymes, and the presence of several active site residues considered typical of the class II enzyme. Hence, the avian enzyme has mixed-class properties, being functionally similar to class I, yet structurally similar to class II, with which it also clusters in phylogenetic trees of characterized vertebrate alcohol dehydrogenases. Comparisons reveal that the class II enzyme is approximately 25% more variable than the "variable" class I enzyme, which itself is more variable than the "constant" class III enzyme. The overall extreme, and the unusual chromatographic behavior may explain why the class II enzyme has previously not been found outside mammals. The properties define a consistent pattern with apparently repeated generation of novel enzyme activities after separate gene duplications. PMID- 7479908 TI - Phage display selection of ligand residues important for Src homology 3 domain binding specificity. AB - The Src homology 3 (SH3) domain is a 50-aa modular unit present in many cellular proteins involved in intracellular signal transduction. It functions to direct protein-protein interactions through the recognition of proline-rich motifs on associated proteins. SH3 domains are important regulatory elements that have been demonstrated to specify distinct regulatory pathways important for cell growth, migration, differentiation, and responses to the external milieu. By the use of synthetic peptides, ligands have been shown to consist of a minimum core sequence and to bind to SH3 domains in one of two pseudosymmetrical orientations, class I and class II. The class I sites have the consensus sequence ZP(L/P)PP psi P whereas the class II consensus is PP psi PPZ (where psi is a hydrophobic residue and Z is a SH3 domain-specific residue). We previously showed by M13 phage display that the Src, Fyn, Lyn, and phosphatidylinositol 3-kinase (PI3K) SH3 domains preferred the same class I-type core binding sequence, RPLPP psi P. These results failed to explain the specificity for cellular proteins displayed by SH3 domains in cells. In the current study, class I and class II core ligand sequences were displayed on the surface of bacteriophage M13 with five random residues placed either N- or C-terminal of core ligand residues. These libraries were screened for binding to the Src, Fyn, Lyn, Yes, and PI3K SH3 domains. By this approach, additional ligand residue preferences were identified that can increase the affinity of SH3 peptide ligands at least 20-fold compared with core peptides. The amino acids selected in the flanking sequences were similar for Src, Fyn, and Yes SH3 domains; however, Lyn and PI3K SH3 domains showed distinct binding specificities. These results indicate that residues that flank the core binding sequences shared by many SH3 domains are important determinants of SH3 binding affinity and selectivity. PMID- 7479909 TI - Multiple calcium channel transcripts in rat osteosarcoma cells: selective activation of alpha 1D isoform by parathyroid hormone. AB - Osteoblasts express calcium channels that are thought to be involved in the transduction of extracellular signals regulating bone metabolism. The molecular identity of the pore-forming subunit (alpha 1) of L-type calcium channel(s) was determined in rat osteosarcoma UMR-106 cells, which express an osteoblast phenotype. A homology-based reverse transcriptase-polymerase chain reaction cloning strategy was employed that used primers spanning the fourth domain. Three types of cDNAs were isolated, corresponding to the alpha 1S (skeletal), alpha 1C (cardiac), and alpha 1D (neuroendocrine) isoforms. In the transmembrane segment IVS3 and the extracellular loop formed by the IVS3-S4 linker, a single pattern of mRNA splicing was found that occurs in all three types of calcium channel transcripts. Northern blot analysis revealed an 8.6-kb mRNA that hybridized to the alpha 1C probe and 4.8- and 11.7-kb mRNAs that hybridized to the alpha 1S and alpha 1D probes. Antisense oligonucleotides directed to the calcium channel alpha 1D transcript, but not those directed to alpha 1S or alpha 1C transcripts, inhibited the rise of intracellular calcium induced by parathyroid hormone. However, alpha 1D antisense oligonucleotides had no effect on the accumulation of cAMP induced by parathyroid hormone. When L-type calcium channels were activated with Bay K 8644, antisense oligonucleotides to each of the three isoforms partially inhibited the rise of intracellular calcium. The present results provide evidence for the expression of three distinct calcium channel alpha 1 subunit isoforms in an osteoblast-like cell line. We conclude that the alpha 1D isoform is selectively activated by parathyroid hormone. PMID- 7479910 TI - Activated Drosophila Ras1 is selectively suppressed by isoprenyl transferase inhibitors. AB - Ras CAAX (C = cysteine, A = aliphatic amino acid, and X = any amino acid) peptidomimetic inhibitors of farnesyl protein transferase suppress Ras-dependent cell transformation by preventing farnesylation of the Ras oncoprotein. These compounds are potential anticancer agents for tumors associated with Ras mutations. The peptidomimetic FTI-254 was tested for Ras1-inhibiting activity in whole animals by injection of activated Ras1val12 Drosophila larvae. FTI-254 decreased the ability of Ras1val12 to form supernumerary R7 photoreceptor cells in the compound eye of transformed flies. In contrast, it had no effect on the related supernumerary R7 phenotypes of flies transformed with either the activated sevenless receptor tyrosine kinase, Raf kinase, or a chimeric Ras1val12 protein that is membrane associated through myristylation instead of isoprenylation. Therefore, FTI-254 acts as an isoprenylation inhibitor to selectively inhibit Ras1val12 signaling activity in a whole-animal model system. PMID- 7479911 TI - Thermodynamic parameters for the helix-coil transition of oligopeptides: molecular dynamics simulation with the peptide growth method. AB - The helix-coil transition equilibrium of polypeptides in aqueous solution was studied by molecular dynamics simulation. The peptide growth simulation method was introduced to generate dynamic models of polypeptide chains in a statistical (random) coil or an alpha-helical conformation. The key element of this method is to build up a polypeptide chain during the course of a molecular transformation simulation, successively adding whole amino acid residues to the chain in a predefined conformation state (e.g., alpha-helical or statistical coil). Thus, oligopeptides of the same length and composition, but having different conformations, can be incrementally grown from a common precursor, and their relative conformational free energies can be calculated as the difference between the free energies for growing the individual peptides. This affords a straightforward calculation of the Zimm-Bragg sigma and s parameters for helix initiation and helix growth. The calculated sigma and s parameters for the polyalanine alpha-helix are in reasonable agreement with the experimental measurements. The peptide growth simulation method is an effective way to study quantitatively the thermodynamics of local protein folding. PMID- 7479912 TI - Muscarinic acetylcholine receptor down-regulation limits the extent of inhibition of cell cycle progression in Chinese hamster ovary cells. AB - Cellular desensitization is believed to be important for growth control but direct evidence is lacking. In the current study we compared effects of wild-type and down-regulation-resistant mutant m3 muscarinic receptors on Chinese hamster ovary (CHO-K1) cell desensitization, proliferation, and transformation. We found that down-regulation of m3 muscarinic acetylcholine receptors was the principal mechanism of desensitization of receptor-activated inositol phosphate phospholipid hydrolysis in these cells. Activation of wild-type and mutant receptors inhibited anchorage-independent growth as assayed by colony formation in agar. However, the potency for inhibition of anchorage-independent growth was greater for cells expressing the mutant receptor. Activation of either receptor also initially inhibited anchorage-dependent cell proliferation in randomly growing populations. Rates of DNA synthesis and cell division were profoundly reduced by carbachol in cells expressing either receptor at early time points. Analysis of cell cycle parameters indicated that cell cycle progression was inhibited at transitions from G1 to S and G2/M to G1 phases. However, mutant receptor effects on anchorage-dependent growth were sustained, whereas wild-type receptor effects were transient. Thus, receptor down-regulation restored cell cycle progression. In contrast, activation of either receptor blocked entry into the cell cycle from quiescence, and this response was not reduced by receptor down-regulation. Therefore, activation of m3 muscarinic acetylcholine receptors inhibited CHO cell anchorage-dependent and -independent growth. In anchored cells carbachol inhibited the cell cycle at three distinct points. Inhibitions at two of these points were eliminated by wild-type receptor down-regulation while the other was not. These results directly demonstrate that desensitization mechanisms can act as principal determinants of cellular growth responses. PMID- 7479913 TI - Anti-nuclear antibody production and immune-complex glomerulonephritis in BALB/c mice treated with pristane. AB - The pathogenesis of systemic lupus erythematosus is thought to be primarily under genetic control, with environmental factors playing a secondary role. However, it has been shown recently that intraperitoneal injection of pristane (2,6,10,14 tetramethylpentadecane) induces autoantibodies typical of lupus in BALB/c mice, a strain not usually considered to be genetically susceptible to the disease. In this study, the induction of autoimmune disease by pristane was investigated. BALB/c mice receiving pristane were tested for autoantibody production and histopathological evidence of glomerulonephritis. Six of 11 mice developed IgM anti-single-stranded DNA antibodies shortly after receiving pristane and 4 developed IgM anti-histone antibodies, but anti-double-stranded DNA antibodies were absent. IgG anti-DNA and anti-histone antibodies were absent. In contrast, the lupus-associated anti-nuclear ribonucleoprotein/Sm and anti-Su autoantibodies produced by these mice were predominantly IgG. In addition to autoantibodies, most of the mice developed significant proteinuria. Light microscopy of the kidney showed segmental or diffuse proliferative glomerulonephritis. Electron microscopy showed subepithelial and mesangial immune-complex deposits and epithelial foot process effacement. Immunofluorescence revealed striking glomerular deposition of IgM, IgG, and C3 with a mesangial or mesangiocapillary distribution. Thus, pristane induces immune-complex glomerulonephritis in association with autoantibodies typical of lupus in BALB/c mice. These data support the idea that lupus is produced by an interplay of genetic and environmental factors and that unlike the MRL or (NZB x W)F1 mouse models, in which genetic susceptibility factors are of primary importance, environmental factors are of considerable importance in the autoimmune disease of pristane treated BALB/c mice. PMID- 7479914 TI - Mice deficient in the orphan receptor steroidogenic factor 1 lack adrenal glands and gonads but express P450 side-chain-cleavage enzyme in the placenta and have normal embryonic serum levels of corticosteroids. AB - The orphan nuclear receptor steroidogenic factor 1 (SF-1) is expressed in the adrenal cortex and gonads and regulates the expression of several P450 steroid hydroxylases in vitro. We examined the role of SF-1 in the adrenal glands and gonads in vivo by a targeted disruption of the mouse SF-1 gene. All SF-1 deficient mice died shortly after delivery. Their adrenal glands and gonads were absent, and persistent Mullerian structures were found in all genotypic males. While serum levels of corticosterone in SF-1-deficient mice were diminished, levels of adrenocorticotropic hormone (ACTH) were elevated, consistent with intact pituitary corticotrophs. Intrauterine survival of SF-1-deficient mice appeared normal, and they had normal serum level of corticosterone and ACTH, probably reflecting transplacental passage of maternal steroids. We tested whether SF-1 is required for P450 side-chain-cleavage enzyme (P450scc) expression in the placenta, which expresses both SF-1 and P450scc, and found that in contrast to its strong activation of the P450scc gene promoter in vitro, the absence of SF-1 had no effect on P450scc mRNA levels in vivo. Although the region targeted by our disruption is shared by SF-1 and by embryonal long terminal repeat-binding protein (ELP), a hypothesized alternatively spliced product, we believe that the observed phenotype reflects absent SF-1 alone, as PCR analysis failed to detect ELP transcripts in any mouse tissue, and sequences corresponding to ELP are not conserved across species. These results confirm that SF-1 is an important regulator of adrenal and gonadal development, but its regulation of steroid hydroxylase expression in vivo remains to be established. PMID- 7479917 TI - Particles move along actin filament bundles in nerve growth cones. AB - Organelle movement along actin filaments has been demonstrated in dissociated squid axoplasm [Kurznetsov, S. A., Langford, G.M. & Weiss, D. G. (1992) Nature (London) 356, 722-725 and Bearer, E.L., DeGiorgis, J.A., Bodner, R.A., Kao, A.W. & Reese, T.S. (1993) Proc. Natl. Acad. Sci. USA 90, 11252-11256] but has not been shown to occur in intact neurons. Here we demonstrate that intracellular transport occurs along actin filament bundles in intact neuronal growth cones. We used video-enhanced differential interference contrast microscopy to observe intracellular transport in superior cervical ganglion neurons cultured under conditions that enhance the visibility of actin bundles within growth cone lamellipodia. Intracellular particles, ranging in size from < 0.5-1.5 microns, moved along linear structures (termed transport bundles) at an average maximum rate of 0.48 micron/sec. After particle movement had been viewed, cultures were preserved by rapid perfusion with chemical fixative. To determine whether particle transport occurred along actin, we then used fluorescence microscopy to correlate this movement with actin and microtubule distributions in the same growth cones. The observed transport bundles colocalized with actin but not with microtubules. The rates of particle movement and the association of moving particles with actin filament bundles suggest that myosins may participate in the transport of organelles (or other materials) in intact neurons. PMID- 7479916 TI - Structure determination of murine mitochondrial carbonic anhydrase V at 2.45-A resolution: implications for catalytic proton transfer and inhibitor design. AB - The three-dimensional structure of murine mitochondrial carbonic anhydrase V has been determined and refined at 2.45-A resolution (crystallographic R factor = 0.187). Significant structural differences unique to the active site of carbonic anhydrase V are responsible for differences in the mechanism of catalytic proton transfer as compared with other carbonic anhydrase isozymes. In the prototypical isozyme, carbonic anhydrase II, catalytic proton transfer occurs via the shuttle group His-64; carbonic anhydrase V has Tyr-64, which is not an efficient proton shuttle due in part to the bulky adjacent side chain of Phe-65. Based on analysis of the structure of carbonic anhydrase V, we speculate that Tyr-131 may participate in proton transfer due to its proximity to zinc-bound solvent, its solvent accessibility, and its electrostatic environment in the protein structure. Finally, the design of isozyme-specific inhibitors is discussed in view of the complex between carbonic anhydrase V and acetazolamide, a transition state analogue. Such inhibitors may be physiologically important in the regulation of blood glucose levels. PMID- 7479915 TI - Coexpression of NF-kappa B/Rel and Sp1 transcription factors in human immunodeficiency virus 1-induced, dendritic cell-T-cell syncytia. AB - Productive infection of T cells with human immunodeficiency virus 1 (HIV-1) typically requires that the T cells be stimulated with antigens or mitogens. This requirement has been attributed to the activation of the transcription factor NF kappa B, which synergizes with the constitutive transcription factor Sp1 to drive the HIV-1 promoter. Recently, we have found that vigorous replication of HIV-1 takes place in nonactivated memory T cells after syncytium formation with dendritic cells (DCs). These syncytia lack activated cells as determined by an absence of staining for Ki-67 cell cycle antigen. The expression and activity of NF-kappa B and Sp1 were, therefore, analyzed in isolated T cells and DCs from humans and mice. We have used immunolabeling, Western blot analysis, and electrophoretic mobility shift and supershift assays. T cells lack active NF kappa B but express Sp1 as expected. DCs express high levels of all known NF kappa B and Rel proteins, with activity residing primarily within RelB, p50, and p65. However, DCs lack Sp1, which may explain the failure of HIV-1 to replicate in purified DCs. Coexpression of NF-kappa B and Sp1 occurs in the heterologous DC T-cell syncytia that are induced by HIV-1. Therefore, HIV-1-induced cell fusion brings together factors that upregulate virus transcription. Since DCs and memory T cells frequently traffic together in situ, these unusual heterologous syncytia could develop in infected individuals and lead to chronic HIV-1 replication without ostensible immune stimulation. PMID- 7479918 TI - Myelin-associated neurite growth-inhibitory proteins and suppression of regeneration of immature mammalian spinal cord in culture. AB - Neurite outgrowth across spinal cord lesions in vitro is rapid in preparations isolated from the neonatal opossum Monodelphis domestica up to the age of 12 days. At this age oligodendrocytes, myelin, and astrocytes develop and regeneration ceases to occur. The role of myelin-associated neurite growth inhibitory proteins, which increase in concentration at 10-13 days, was investigated in culture by applying the antibody IN-1, which blocks their effects. In the presence of IN-1, 22 out of 39 preparations from animals aged 13 17 days showed clear outgrowth of processes into crushes. When 34 preparations from 13-day-old animals were crushed and cultured without antibody, no axons grew into the lesion. The success rate with IN-1 was comparable to that seen in younger animals but the outgrowth was less profuse. IN-1 was shown by immunocytochemistry to penetrate the spinal cord. Other antibodies which penetrated the 13-day cord failed to promote fiber outgrowth. To distinguish between regeneration by cut neurites and outgrowth by developing uncut neurites, fibers in the ventral fasciculus were prelabeled with carbocyanine dyes and subsequently injured. The presence of labeled fibers in the lesion indicated that IN-1 promoted regeneration. These results show that the development of myelin associated growth-inhibitory proteins contributes to the loss of regeneration as the mammalian central nervous system matures. The definition of a critical period for regeneration, coupled with the ability to apply trophic as well as inhibitory molecules to the culture, can permit quantitative assessment of molecular interactions that promote spinal cord regeneration. PMID- 7479919 TI - Rotation of subunits during catalysis by Escherichia coli F1-ATPase. AB - During oxidative and photo-phosphorylation, F0F1-ATP synthases couple the movement of protons down an electrochemical gradient to the synthesis of ATP. One proposed mechanistic feature that has remained speculative is that this coupling process requires the rotation of subunits within F0F1. Guided by a recent, high resolution structure for bovine F1 [Abrahams, J. P., Leslie, A. G., Lutter, R. & Walker, J. E. (1994) Nature (London) 370, 621-628], we have developed a critical test for rotation of the central gamma subunit relative to the three catalytic beta subunits in soluble F1 from Escherichia coli. In the bovine F1 structure, a specific point of contact between the gamma subunit and one of the three catalytic beta subunits includes positioning of the homolog of E. coli gamma subunit C87 (gamma C87) close to the beta-subunit 380DELSEED386 sequence. A beta D380C mutation allowed us to induce formation of a specific disulfide bond between beta and gamma C87 in soluble E. coli F1. Formation of the crosslink inactivated beta D380C-F1, and reduction restored full activity. Using a dissociation/reassembly approach with crosslinked beta D380C-F1, we incorporated radiolabeled beta subunits into the two noncrosslinked beta-subunit positions of F1. After reduction of the initial nonradioactive beta-gamma crosslink, only exposure to conditions for catalytic turnover results in similar reactivities of unlabeled and radiolabeled beta subunits with gamma C87 upon reoxidation. The results demonstrate that gamma subunit rotates relative to the beta subunits during catalysis. PMID- 7479920 TI - Lyophilization-induced reversible changes in the secondary structure of proteins. AB - Changes in the secondary structure of some dozen different proteins upon lyophilization of their aqueous solutions have been investigated by means of Fourier-transform infrared spectroscopy in the amide III band region. Dehydration markedly (but reversibly) alters the secondary structure of all the proteins studied, as revealed by both the quantitative analysis of the second derivative spectra and the Gaussian curve fitting of the original infrared spectra. Lyophilization substantially increases the beta-sheet content and lowers the alpha-helix content of all proteins. In all but one case, proteins become more ordered upon lyophilization. PMID- 7479921 TI - Reactivated and latent varicella-zoster virus in human dorsal root ganglia. AB - Ganglia obtained at autopsy were examined by in situ hybridization from one patient with zoster (also called herpes zoster or shingles), two varicella-zoster virus (VZV)-seropositive patients with clinical evidence of zoster, one VZV seronegative child, and one fetus. Ganglia positive for VZV had a hybridization signal in both neuronal and nonneuronal satellite cells. Ganglia obtained from the fetus and from the seronegative infant were consistently negative for VZV. Two striking observations were evident regarding the presence of VZV DNA in ganglia obtained from the individual with zoster at the time of death. First, ganglia innervating the sites of reactivation and ganglia innervating adjacent sites yielded strongly positive signals in neurons and satellite cells, whereas ganglia from distant sites were rarely positive. Second, VZV DNA was found in both the nuclei and the cytoplasm of neurons innervating areas of zoster. However, in neurons innervating zoster-free areas, VZV DNA was found only in the nucleus of neurons and their supporting satellite cells. Immunohistochemistry with a fluorescent monoclonal antibody to the VZV glycoprotein gpI, a late virus protein, revealed a positive signal in the cytoplasm of ganglia with clinical evidence of reactivation. These results illustrate that both neuronal and satellite cells become latently infected following primary VZV infection. The presence of VZV DNA and gpI in the cytoplasm of neurons demonstrates productive infection following reactivation at the site of latency. PMID- 7479922 TI - Interleukin 2 induces CD8+ T cell-mediated suppression of human immunodeficiency virus replication in CD4+ T cells and this effect overrides its ability to stimulate virus expression. AB - The nonlytic suppression of human immunodeficiency virus (HIV) production from infected CD4+ T cells by CD8+ lymphocytes from HIV-infected individuals is one of the most potent host-mediated antiviral activities observed in vitro. We demonstrate that the pleiotropic cytokine interleukin 2 (IL-2), but not IL-12, is a potent inducer of the CD8+ HIV suppressor phenomenon. IL-2 induces HIV expression in peripheral blood or lymph node mononuclear cells from HIV-infected individuals in the absence of CD8+ T cells. However, IL-2 induces CD8+ T cells to suppress HIV expression when added back to these cultures, and this effect dramatically supersedes the ability to IL-2 to induce HIV expression. Five to 25 times fewer CD8+ cells were required to obtain comparable levels of inhibition of viral production if they were activated in the presence of IL-2 as compared with IL-12 or no exogenous cytokine. Furthermore, IL-2 appeared either to induce a qualitative increase in HIV suppressor cell activity or to increase the relative frequency of suppressor cells in the activated (CD25+) CD8+ populations. Analyses of proviral levels in peripheral blood mononuclear cells suggest that CD8+ T cell mediated lysis of in vivo infected cells is not induced by IL-2. These results have implications for our understanding of the effects of impaired IL-2 production during HIV disease as well as the overall effects of IL-2-based immunotherapy on HIV replication in vivo. PMID- 7479923 TI - Down-regulation of NF-kappa B protein levels in activated human lymphocytes by 1,25-dihydroxyvitamin D3. AB - The effect of 1,25-dihydroxyvitamin D3 [1,25(OH)2)D3], a steroid hormone with immunomodulating properties, on nuclear factor kappa B (NF-kappa B) proteins was examined in in vitro activated normal human lymphocytes by Western blot analysis. Over a 72-hr period of activation, the expression of the 50-kDa NF-kappa B, p50, and its precursor, p105, was increased progressively. When cells were activated in the presence of 1,25(OH)2D3, the levels of the mature protein as well as its precursor were decreased. The effect of the hormone on the levels of p50 was demonstrable in the cytosolic and nuclear compartments; it required between 4 and 8 hr and was specific, as 25-hydroxyvitamin D3 and 24,25-dihydroxyvitamin D3 were ineffective. Besides p50, 1,25(OH)2D3 decreased the levels of another NF-kappa B protein, namely c-rel. In addition, 1,25(OH)2D3 decreased the abundance of a specific DNA-protein complex formed upon incubation of nuclear extracts from activated lymphocytes with a labeled NF-kappa B DNA binding motif. Further, 1,25(OH)2D3 inhibited the transcriptional activity of NF-kappa B in Jurkat cells transiently transfected with a construct containing four tandem repeats of the NF kappa B binding sequence of the immunoglobulin kappa light chain gene linked to the chloramphenicol acetyltransferase reporter gene. These observations demonstrate directly that there is de novo synthesis of NF-kappa B during human lymphocyte activation and suggest that this process is hormonally regulated. PMID- 7479925 TI - Blocking effects of polyunsaturated fatty acids on Na+ channels of neonatal rat ventricular myocytes. AB - Recent evidence indicates that polyunsaturated long-chain fatty acids (PUFAs) prevent lethal ischemia-induced cardiac arrhythmias in animals and probably in humans. To increase understanding of the mechanism(s) of this phenomenon, the effects of PUFAs on Na+ currents were assessed by the whole-cell patch-clamp technique in cultured neonatal rat ventricular myocytes. Extracellular application of the free 5,8,11,14,17-eicosapentaenoic acid (EPA) produced a concentration-dependent suppression of ventricular, voltage-activated Na+ currents (INa). After cardiac myocytes were treated with 5 or 10 microM EPA, the peak INa (elicited by a single-step voltage change with pulses from -80 to -30 mV) was decreased by 51% +/- 8% (P < 0.01; n = 10) and 64% +/- 5% (P < 0.001; n = 21), respectively, within 2 min. Likewise, the same concentrations of 4,7,10,16,19-docosahexaenoic acid produced the same inhibition of INa. By contrast, 5 and 10 microM arachidonic acid (AA) caused less inhibition of INa, but both n - 6 and n - 3 PUFAs inhibited INa significantly. A monounsaturated fatty acid and a saturated fatty acid did not. After washing out EPA, INa returned to the control level. Raising the concentration of EPA to 40 microM completely blocked INa. The IC50 of EPA was 4.8 microM. The inhibition of this Na+ channel was found to be dose and time, but not use dependent. Also, the EPA induced inhibition of INa was voltage dependent, since 10 microM EPA produced 83% +/- 7% and 29% +/- 5% inhibition of INa elicited by pulses from -80 to -30 mV and from -150 to -30 mV, respectively, in single-step voltage changes. A concentration of 10 microM EPA shifted the steady-state inactivation curve of INa by -19 +/- 3 mV (n = 7; P < 0.01). These effects of PUFAs on INa may be important for their antiarrhythmic effect in vivo. PMID- 7479926 TI - Differential activation of yeast adenylyl cyclase by Ras1 and Ras2 depends on the conserved N terminus. AB - Although both Ras1 and Ras2 activate adenylyl cyclase in yeast, a number of differences can be observed regarding their function in the cAMP pathway. To explore the relative contribution of conserved and variable domains in determining these differences, chimeric RAS1-RAS2 or RAS2-RAS1 genes were constructed by swapping the sequences encoding the variable C-terminal domains. These constructs were expressed in a cdc25ts ras1 ras2 strain. Biochemical data show that the difference in efficacy of adenylyl cyclase activation between the two Ras proteins resides in the highly conserved N-terminal domain. This finding is supported by the observation that Ras2 delta, in which the C-terminal domain of Ras2 has been deleted, is a more potent activator of the yeast adenylyl cyclase than Ras1 delta, in which the C-terminal domain of Ras1 has been deleted. These observations suggest that amino acid residues other than the highly conserved residues of the effector domain within the N terminus may determine the efficiency of functional interaction with adenylyl cyclase. Similar levels of intracellular cAMP were found in Ras1, Ras1-Ras2, Ras1 delta, Ras2, and Ras2-Ras1 strains throughout the growth curve. This was found to result from the higher expression of Ras1 and Ras1-Ras2, which compensate for their lower efficacy in activating adenylyl cyclase. These results suggest that the difference between the Ras1 and the Ras2 phenotype is not due to their different efficacy in activating the cAMP pathway and that the divergent C-terminal domains are responsible for these differences, through interaction with other regulatory elements. PMID- 7479924 TI - Staphylococcal enterotoxins modulate interleukin 2 receptor expression and ligand induced tyrosine phosphorylation of the Janus protein-tyrosine kinase 3 (Jak3) and signal transducers and activators of transcription (Stat proteins). AB - Staphylococcal enterotoxins (SE) stimulate T cells expressing the appropriate variable region beta chain of (V beta) T-cell receptors and have been implicated in the pathogenesis of several autoimmune diseases. Depending on costimulatory signals, SE induce either proliferation or anergy in T cells. In addition, SE can induce an interleukin-2 (IL-2) nonresponsive state and apoptosis. Here, we show that SE induce dynamic changes in the expression of and signal transduction through the IL-2 receptor (IL-2R) beta and gamma chains (IL-2R beta and IL-2R gamma) in human antigen-specific CD4+ T-cell lines. Thus, after 4 hr of exposure to SEA and SEB, the expression of IL-2R beta was down-regulated, IL-2R gamma was slightly up-regulated, while IL-2R alpha remained largely unaffected. The changes in the composition of IL-2Rs were accompanied by inhibition of IL-2-induced tyrosine phosphorylation of the Janus protein-tyrosine kinase 3 (Jak3) and signal transducers and activators of transcription called Stat3 and Stat5. In parallel experiments, IL-2-driven proliferation was inhibited significantly. After 16 hr of exposure to SE, the expression of IL-2R beta remained low, while that of IL2R alpha and IL2R gamma was further up-regulated, and ligand-induced tyrosine phosphorylation of Jak3 and Stat proteins was partly normalized. Yet, IL-2-driven proliferation remained profoundly inhibited, suggesting that signaling events other than Jak3/Stat activation had also been changed following SE stimulation. In conclusion, our data suggest that SE can modulate IL-2R expression and signal transduction involving the Jak/Stat pathway in CD4+ T-cell lines. PMID- 7479927 TI - Identification of a unique membrane-bound molecule on a hemopoietic stem cell line and on multipotent progenitor cells. AB - Hemopoietic stem cells are a distinct population of cells that can differentiate into multilineages of hemopoietic cells and have long-term repopulation capability. A few membrane-bound molecules have been found to be preferentially, but not uniquely, present on the surface of these primitive cells. We report here the identification of a unique 105-kDa glycoprotein on the surface of hemopoietic stem cell line BL3. This molecule, recognized by the absorbed antiserum, is not present on the surface of myeloid progenitors 32D and FDC-P1 cells, EL4 T cells, and NIH 3T3 fibroblasts. This antiserum can also be used to block the proliferation of BL3 cells even in the presence of mitogen-stimulated spleen cell conditioned medium, which is known to have a stimulating activity on BL3 cells. It can also inhibit development of in vitro, fetal liver cell-derived multilineage colonies, but not other types of colonies, and of in vivo bone marrow cell-derived colony-forming unit spleen foci. These data suggest that gp105 plays an important role in hemopoietic stem cell differentiation. PMID- 7479928 TI - Mismatch repair in Escherichia coli enhances instability of (CTG)n triplet repeats from human hereditary diseases. AB - Long CTG triplet repeats which are associated with several human hereditary neuromuscular disease genes are stabilized in ColE1-derived plasmids in Escherichia coli containing mutations in the methyl-directed mismatch repair genes (mutS, mutL, or mutH). When plasmids containing (CTG)180 were grown for about 100 generations in mutS, mutL, or mutH strains, 60-85% of the plasmids contained a full-length repeat, whereas in the parent strain only about 20% of the plasmids contained the full-length repeat. The deletions occur only in the (CTG)180 insert, not in DNA flanking the repeat. While many products of the deletions are heterogeneous in length, preferential deletion products of about 140, 100, 60, and 20 repeats were observed. We propose that the E. coli mismatch repair proteins recognize three-base loops formed during replication and then generate long single-stranded gaps where stable hairpin structures may form which can be bypassed by DNA polymerase during the resynthesis of duplex DNA. Similar studies were conducted with plasmids containing CGG repeats; no stabilization of these triplets was found in the mismatch repair mutants. Since prokaryotic and human mismatch repair proteins are similar, and since several carcinoma cell lines which are defective in mismatch repair show instability of simple DNA microsatellites, these mechanistic investigations in a bacterial cell may provide insights into the molecular basis for some human genetic diseases. PMID- 7479929 TI - A recombinant bcl-x s adenovirus selectively induces apoptosis in cancer cells but not in normal bone marrow cells. AB - Many cancers overexpress a member of the bcl-2 family of inhibitors of apoptosis. To determine the role of these proteins in maintaining cancer cell viability, an adenovirus vector that expresses bcl-xs, a functional inhibitor of these proteins, was constructed. Even in the absence of an exogenous apoptotic signal such as x-irradiation, this virus specifically and efficiently kills carcinoma cells arising from multiple organs including breast, colon, stomach, and neuroblasts. In contrast, normal hematopoietic progenitor cells and primitive cells capable of repopulating severe combined immunodeficient mice were refractory to killing by the bcl-xs adenovirus. These results suggest that Bcl-2 family members are required for survival of cancer cells derived from solid tissues. The bcl-xs adenovirus vector may prove useful in killing cancer cells contaminating the bone marrow of patients undergoing autologous bone marrow transplantation. PMID- 7479930 TI - Selective neuronal toxicity of cocaine in embryonic mouse brain cocultures. AB - Cocaine exposure in utero causes severe alterations in the development of the central nervous system. To study the basis of these teratogenic effects in vitro, we have used cocultures of neurons and glial cells from mouse embryonic brain. Cocaine selectively affected embryonic neuronal cells, causing first a dramatic reduction of both number and length of neurites and then extensive neuronal death. Scanning electron microscopy demonstrated a shift from a multipolar neuronal pattern towards bi- and unipolarity prior to the rounding up and eventual disappearance of the neurons. Selective toxicity of cocaine on neurons was paralleled by a concomitant decrease of the culture content in microtubule associated protein 2 (MAP2), a neuronal marker measured by solid-phase immunoassay. These effects on neurons were reversible when cocaine was removed from the culture medium. In contrast, cocaine did not affect astroglial cells and their glial fibrillary acidic protein (GFAP) content. Thus, in embryonic neuronal glial cell cocultures, cocaine induces major neurite perturbations followed by neuronal death without affecting the survival of glial cells. Provided similar neuronal alterations are produced in the developing human brain, they could account for the qualitative or quantitative defects in neuronal pathways that cause a major handicap in brain function following in utero exposure to cocaine. PMID- 7479932 TI - Sperm capacitation in humans is transient and correlates with chemotactic responsiveness to follicular factors. AB - In humans, only a small fraction (2-12%) of a sperm population can respond by chemoattraction to follicular factors. This recent finding led to the hypothesis that chemotaxis provides a mechanism for selective recruitment of functionally mature spermatozoa (i.e., of capacitated spermatozoa, which possess the potential to undergo the acrosome reaction and fertilize the egg). This study aimed to examine this possibility. Capacitated spermatozoa were identified by their ability to undergo the acrosome reaction upon stimulation with phorbol 12 myristate 13-acetate. Under capacitating conditions, only a small portion (2-14%) of the spermatozoa were found to be capacitated. The spermatozoa were then separated according to their chemotactic activity, which resulted in a subpopulation enriched with chemotactically responsive spermatozoa and a subpopulation depleted of such spermatozoa. The level of capacitated spermatozoa in the former was approximately 13-fold higher than that in the latter. The capacitated state was temporary (50 min < life span < 240 min), and it was synchronous with the chemotactic activity. A continuous process of replacement of capacitated/chemotactic spermatozoa within a sperm population was observed. Spermatozoa that had stopped being capacitated did not become capacitated again, which indicates that the capacitated state is acquired only once in a sperm's lifetime. A total sperm population depleted of capacitated spermatozoa stopped being chemotactic. When capacitated spermatozoa reappeared, chemotactic activity was restored. These observations suggest that spermatozoa acquire their chemotactic responsiveness as part of the capacitation process and lose this responsiveness when the capacitated state is terminated. We suggest that the role of sperm chemotaxis in sperm-egg interaction in vivo may indeed be selective recruitment of capacitated spermatozoa for fertilizing the egg. PMID- 7479934 TI - Synergism between UV-B radiation and a pathogen magnifies amphibian embryo mortality in nature. AB - Previous research has shown that amphibians have differential sensitivity to ultraviolet-B (UV-B) radiation. In some species, ambient levels of UV-B radiation cause embryonic mortality in nature. The detrimental effects of UV-B alone or with other agents may ultimately affect amphibians at the population level. Here, we experimentally demonstrate a synergistic effect between UV-B radiation and a pathogenic fungus in the field that increases the mortality of amphibian embryos compared with either factor alone. Studies investigating single factors for causes of amphibian egg mortality or population declines may not reveal the complex factors involved in declines. PMID- 7479933 TI - Peptide conjugation to an in vitro-selected DNA ligand improves enzyme inhibition. AB - An in vitro selection technique was used to identify a specific high-affinity DNA ligand targeted to human neutrophil elastase (HNE). 1H NMR data and a comparative analysis of the selected sequences suggest that the DNA folds into a G-quartet structure with duplexed ends. The high-affinity binding DNA alone did not inhibit the enzymatic activity of HNE. The DNA was covalently attached to a tetrapeptide, N-methoxysuccinyl-Ala-Ala-Pro-Val, that is a weak competitive inhibitor of HNE. HNE was inhibited by this DNA-peptide conjugate nearly five orders of magnitude more effectively than by the peptide alone. These results demonstrate that in vitro-selected nucleic acids can be used as a vehicle for molecular delivery. PMID- 7479931 TI - Cloning of a sodium channel alpha subunit from rabbit Schwann cells. AB - Overlapping cDNA clones spanning the entire coding region of a Na-channel alpha subunit were isolated from cultured Schwann cells from rabbits. The coding region predicts a polypeptide (Nas) of 1984 amino acids exhibiting several features characteristic of Na-channel alpha subunits isolated from other tissues. Sequence comparisons showed that the Nas alpha subunit resembles most the family of Na channels isolated from brain (approximately 80% amino acid identity) and is least similar (approximately 55% amino acid identity) to the atypical Na channel expressed in human heart and the partial rat cDNA, NaG. As for the brain II and III isoforms, two variants of Nas exist that appear to arise by alternative splicing. The results of reverse transcriptase-polymerase chain reaction experiments suggest that expression of Nas transcripts is restricted to cells in the peripheral and central nervous systems. Expression was detected in cultured Schwann cells, sciatic nerve, brain, and spinal cord but not in skeletal or cardiac muscle, liver, kidney, or lung. PMID- 7479935 TI - The genetic link between the Chinese bamboo partridge (Bambusicola thoracica) and the chicken and junglefowls of the genus Gallus. AB - Further comparison of mitochondrial control-region DNA base sequences of 16 avian species belonging to the subfamily Phasianinae revealed the following: (i) Generalized perdicine birds (quails and partridges) are of ancient lineages. Even the closest pair, the common quail (Coturnix coturnix japonica) and the Chinese bamboo partridge (Bambusicola thoracica), maintained only 85.71% identity. (ii) The 12 species of phasianine birds previously and presently studied belonged to three distinct branches. The first branch was made exclusively of members of the genus Gallus, while the second branch was made of pheasants of the genera Phasianus, Chrysolophus, and Syrmaticus. Gallopheasants of the genus Lophura were distant cousins to these pheasants. The great argus (Argusianus argus) and peafowls of the genus Pavo constituted the third branch. The position of peacock pheasants of the genus Polyplectron in the third branch was similar to that of the genus Lophura in the second branch. Members of the fourth phasianine branch, such as tragopans and monals, were not included in the present study. (iii) The one perdicine species, Bambusicola thoracica, was more closely related to phasianine genera Gallus and Pavo than to members of other perdicine genera. The above might indicate that Bambusicola belong to one-stem perdicine lineage that later splits into two sublineages that yielded phasianine birds, one evolving to Gallus, and the other differentiating toward Pavo and its allies. PMID- 7479936 TI - The gene distribution of the maize genome. AB - Previous investigations from our laboratory showed that the genomes of plants, like those of vertebrates, are mosaics of isochores, i.e., of very long DNA segments that are compositionally homogeneous and that can be subdivided into a small number of families characterized by different GC levels (GC is the mole fraction of guanine+cytosine). Compositional DNA fractions corresponding to different isochore families were used to investigate, by hybridization with appropriate probes, the gene distribution in vertebrate genomes. Here we report such a study on the genome of a plant, maize. The gene distribution that we found is most striking, in that almost all genes are present in isochores covering an extremely narrow (1-2%) GC range and only representing 10-20% of the genome. This gene distribution, which seems to characterize other Gramineae as well, is remarkably different from the gene distribution previously found in vertebrate genomes. PMID- 7479937 TI - Fine specificity of the antibody response to myelin basic protein in the central nervous system in multiple sclerosis: the minimal B-cell epitope and a model of its features. AB - T cells, B cells, and antibody are found in the white matter of the central nervous system in multiple sclerosis. The epitope center for the antibody response to human myelin basic protein (MBP) fits precisely the minimal epitope Pro85-Val-Val-His-Phe-Phe-Lys-Asn-Ile-Val-Thr-Pro96 for that reported for HLA DR2b (DRB1*1501)-restricted T cells that recognize MBP [Wucherpfenning, K.W., Sette, A., Southwood, S., Oseroff, C., Matsui, M., Strominger, J. & Hafler, D. (1994) J. Exp. Med. 179, 279-290], and overlaps with the reported DR2a-restricted epitope for T cells reactive to MBP [Martin, R., Howell, M. D., Jaraquemada, D., Furlage, M., Richert, J., Brostoff, S., Long, E. O., McFarlin, D. E. & McFarland, H. F. (1991) J. Exp. Med. 173, 19-24]. We describe a molecular model of this epitope. PMID- 7479939 TI - The extent of heterocellular communication mediated by gap junctions is predictive of bystander tumor cytotoxicity in vitro. AB - Herpes simplex virus thymidine kinase (HSV-tk)/ganciclovir (GCV) viral-directed enzyme prodrug gene therapy causes potent, tumor-selective cytotoxicity in animal models in which HSV-tk gene transduction is limited to a minority of tumor cells. The passage of toxic molecules from HSV-tk+ cells to neighboring HSV-tk- cells during GCV therapy is one mechanism that may account for this "bystander" cytotoxicity. To investigate whether gap junction-mediated intercellular coupling could mediate this bystander effect, we used a flow cytometry assay to quantitate the extent of heterocellular coupling between HSV-tk+ murine fibroblasts and both rodent and human tumor cell lines. Bystander tumor cytotoxicity during GCV treatment in a coculture assay was highly correlated (P < 0.001) with the extent of gap junction-mediated coupling. These findings show that gap junction-mediated intercellular coupling contributes to the in vitro bystander effect during HSV tk/GCV therapy and that retroviral transduction of tumor cells is not required for bystander cytotoxicity. PMID- 7479938 TI - Unimpaired autoreactive T-cell traffic within the central nervous system during tumor necrosis factor receptor-mediated inhibition of experimental autoimmune encephalomyelitis. AB - The critical role of tumor necrosis factor (TNF) as a mediator in autoimmune inflammatory processes is evident from in vivo studies with TNF-blocking agents. However, the mechanisms by which TNF, and possibly also its homologue lymphotoxin alpha, contributes to development of pathology in rheumatoid arthritis and Crohn disease and in animal models like experimental autoimmune encephalomyelitis is unclear. Possibilities include regulation of vascular adhesion molecules enabling leukocyte movement into tissues or direct cytokine-mediated effector functions such as mediation of tissue damage. Here we show that administration of a TNF receptor (55 kDa)-IgG fusion protein prevented clinical signs of actively induced experimental autoimmune encephalomyelitis. Significantly, the total number of CD4+ T lymphocytes isolated from the central nervous system of clinically healthy treated versus diseased control animals was comparable. By using a CD45 congenic model of passively transferred experimental autoimmune encephalomyelitis to enable tracking of myelin basic protein-specific effector T lymphocytes, prevention of clinical signs of disease was again demonstrated in treated animals but without quantitative or qualitative impediment to the movement of autoreactive T lymphocytes to and within the central nervous system. Thus, despite the uninterrupted movement of specific T lymphocytes into the target tissue, subsequent disease development was blocked. This provides compelling evidence for a direct effector role of TNF/lymphotoxin alpha in autoimmune tissue damage. PMID- 7479940 TI - Designing safer chemicals: predicting the rates of metabolism of halogenated alkanes. AB - A computational model is presented that can be used as a tool in the design of safer chemicals. This model predicts the rate of hydrogen-atom abstraction by cytochrome P450 enzymes. Excellent correlations between biotransformation rates and the calculated activation energies (delta Hact) of the cytochrome P450 mediated hydrogen-atom abstractions were obtained for the in vitro biotransformation of six halogenated alkanes (1-fluoro-1,1,2,2-tetrachloroethane, 1,1-difluoro-1,2,2-trichloroethane, 1,1,1-trifluro-2,2-dichloroethane, 1,1,1,2 tetrafluoro-2-chloroethane, 1,1,1,2,2,-pentafluoroethane, and 2-bromo-2-chloro 1,1,1-trifluoroethane) with both rat and human enzyme preparations: In(rate, rat liver microsomes) = 44.99 - 1.79(delta Hact), r2 = 0.86; In(rate, human CYP2E1) = 46.99 - 1.77(delta Hact), r2 = 0.97 (rates are in nmol of product per min per nmol of cytochrome P450 and energies are in kcal/mol). Correlations were also obtained for five inhalation anesthetics (enflurane, sevoflurane, desflurane, methoxyflurane, and isoflurane) for both in vivo and in vitro metabolism by humans: In[F(-)]peak plasma = 42.87 - 1.57(delta Hact), r2 = 0.86. To our knowledge, these are the first in vivo human metabolic rates to be quantitatively predicted. Furthermore, this is one of the first examples where computational predictions and in vivo and in vitro data have been shown to agree in any species. The model presented herein provides an archetype for the methodology that may be used in the future design of safer chemicals, particularly hydrochlorofluorocarbons and inhalation anesthetics. PMID- 7479941 TI - Identification and characterization of an immunophilin expressed during the clonal expansion phase of adipocyte differentiation. AB - Mouse 3T3-L1 cells differentiate into fat-laden adipocytes in response to a cocktail of adipogenic hormones. This conversion process occurs in two discrete steps. During an early clonal expansion phase, confluent 3T3-L1 cells proliferate and express the products of the beta and delta members of the CCAAT/enhancer binding protein (C/EBP) family of transcription factors. The cells subsequently arrest mitotic growth, induce the expression of the alpha form of C/EBP, and acquire the morphology of fully differentiated adipocytes. Many of the genes induced during the terminal phase of adipocyte conversion are directly activated by C/EBP alpha, and gratuitous expression of this transcription factor is capable of catalyzing adipose conversion in a number of different cultured cell lines. The genetic program undertaken during the clonal expansion phase of 3T3-L1 differentiation, controlled in part by C/EBP beta and C/EBP delta, is less clearly understood. To study the molecular events occurring during clonal expansion, we have identified mRNAs that selectively accumulate during this phase of adipocyte conversion. One such mRNA encodes an immunophilin hereby designated FKBP51. In this report we provide the initial molecular characterization of FKBP51. PMID- 7479942 TI - Rapamycin inhibits clonal expansion and adipogenic differentiation of 3T3-L1 cells. AB - Differentiating 3T3-L1 cells express an immunophilin early during the adipocyte conversion program as described in this issue [Yeh, W.-C., Li, T.-K., Bierer, B. E. & McKnight, S. L. (1995) Proc. Natl. Acad. Sci. USA 92, 11081-11085]. The temporal expression profile of this protein, designated FK506-binding protein (FKBP) 51, is concordant with the clonal-expansion period undertaken by 3T3-L1 cells after exposure to adipogenic hormones. Having observed FKBP51 synthesis early during adipogenesis, we tested the effects of three immunosuppressive drugs -cyclosporin A, FK506, and rapamycin--on the terminal-differentiation process. Adipocyte conversion was not affected by either cyclosporin A or FK506 and yet was significantly reduced by rapamycin at drug concentrations as low as 10 nM. Clonal expansion was impeded in drug-treated cultures, as was the accumulation of cytoplasmic lipid droplets normally seen late during differentiation. Rapamycin treatment likewise inhibited the expression of CCAAT/enhancer binding protein alpha, a transcription factor required for 3T3-L1 cell differentiation. All three of these effects were reversed by high FK506 concentrations, indicating that the operative inhibitory event was mediated by an immunophilin-rapamycin complex. PMID- 7479943 TI - Human naive and memory T lymphocytes differ in telomeric length and replicative potential. AB - The present study has assessed the replicative history and the residual replicative potential of human naive and memory T cells. Telomeres are unique terminal chromosomal structures whose length has been shown to decrease with cell division in vitro and with increased age in vivo for human somatic cells. We therefore assessed telomere length as a measure of the in vivo replicative history of naive and memory human T cells. Telomeric terminal restriction fragments were found to be 1.4 +/- 0.1 kb longer in CD4+ naive T cells than in memory cells from the same donors, a relationship that remained constant over a wide range of donor age. These findings suggest that the differentiation of memory cells from naive precursors occurs with substantial clonal expansion and that the magnitude of this expansion is, on average, similar over a wide range of age. In addition, when replicative potential was assessed in vitro, it was found that the capacity of naive cells for cell division was 128-fold greater as measured in mean population doublings than the capacity of memory cells from the same individuals. Human CD4+ naive and memory cells thus differ in in vivo replicative history, as reflected in telomeric length, and in their residual replicative capacity. PMID- 7479944 TI - Restriction-modification systems as genomic parasites in competition for specific sequences. AB - Restriction-modification (RM) systems are believed to have evolved to protect cells from foreign DNA. However, this hypothesis may not be sufficient to explain the diversity and specificity in sequence recognition, as well as other properties, of these systems. We report that the EcoRI restriction endonuclease modification methylase (rm) gene pair stabilizes plasmids that carry it and that this stabilization is blocked by an RM of the same sequence specificity (EcoRI or its isoschizomer, Rsr I) but not by an RM of a different specificity (PaeR7I) on another plasmid. The PaeR7I rm likewise stabilizes plasmids, unless an rm gene pair with identical sequence specificity is present. Our analysis supports the following model for stabilization and incompatibility: the descendants of cells that have lost an rm gene pair expose the recognition sites in their chromosomes to lethal attack by any remaining restriction enzymes unless modification by another RM system of the same specificity protects these sites. Competition for specific sequences among these selfish genes may have generated the great diversity and specificity in sequence recognition among RM systems. Such altruistic suicide strategies, similar to those found in virus-infected cells, may have allowed selfish RM systems to spread by effectively competing with other selfish genes. PMID- 7479945 TI - Mouse model for Usher syndrome: linkage mapping suggests homology to Usher type I reported at human chromosome 11p15. AB - Usher syndrome is a group of diseases with autosomal recessive inheritance, congenital hearing loss, and the development of retinitis pigmentosa, a progressive retinal degeneration characterized by night blindness and visual field loss over several decades. The causes of Usher syndrome are unknown and no animal models have been available for study. Four human gene sites have been reported, suggesting at least four separate forms of Usher syndrome. We report a mouse model of type I Usher syndrome, rd5, whose linkage on mouse chromosome 7 to Hbb and tub has homology to human Usher I reported on human chromosome 11p15. The electroretinogram in homozygous rd5/rd5 mouse is never normal with reduced amplitudes that extinguish by 6 months. Auditory-evoked response testing demonstrates increased hearing thresholds more than control at 3 weeks of about 30 decibels (dB) that worsen to about 45 dB by 6 months. PMID- 7479946 TI - Antagonism of WT1 activity by protein self-association. AB - Germline loss-of-function mutations at the Wilms tumor (WT) suppressor locus WT1 are associated with a predisposition to WTs and mild genital system anomalies. In contrast, germ-line missense mutations within the WT1 gene encoding the DNA binding domain often yield a more severe phenotype consisting of WT, sexual ambiguity, and renal nephropathy. In this report, we demonstrate that the products of mutant alleles that impair DNA recognition can antagonize WT1 mediated transcriptional repression. We demonstrate that WT1 can self-associate in vitro and in vivo and that the responsible domain maps to the amino-terminal region of the protein. Oligomers of full-length protein form less efficiently or produce less stable complexes than oligomers between truncated polypeptides and full-length protein. Our data suggest a molecular mechanism to explain how WT1 mutations may act in deregulating cellular proliferation and differentiation. PMID- 7479947 TI - Identification of a putative estrogen response element in the gene encoding brain derived neurotrophic factor. AB - We have been studying the role and mechanism of estrogen action in the survival and differentiation of neurons in the basal forebrain and its targets in the cerebral cortex, hippocampus, and olfactory bulb. Previous work has shown that estrogen-target neurons in these regions widely coexpress the mRNAs for the neurotrophin ligands and their receptors, suggesting a potential substrate for estrogen-neurotrophin interactions. Subsequent work indicated that estrogen regulates the expression of two neurotrophin receptor mRNAs in prototypic peripheral neural targets of nerve growth factor. We report herein that the gene encoding the neurotophin brain-derived neurotrophic factor (BDNF) contains a sequence similar to the canonical estrogen response element found in estrogen target genes. Gel shift and DNA footprinting assays indicate that estrogen receptor-ligand complexes bind to this sequence in the BDNF gene. In vivo, BDNF mRNA was rapidly up-regulated in the cerebral cortex and the olfactory bulb of ovariectomized animals exposed to estrogen. These data suggest that estrogen may regulate BDNF transcription, supporting our hypothesis that estrogen may be in a position to influence neurotrophin-mediated cell functioning, by increasing the availability of specific neurotrophins in forebrain neurons. PMID- 7479948 TI - Artificial regulation of gene expression in Escherichia coli by RNase P. AB - Plasmids encoding various external guide sequences (EGSs) were constructed and inserted into Escherichia coli. In strains harboring the appropriate plasmids, the expression of fully induced beta-galactosidase and alkaline phosphatase activity was reduced by more than 50%, while no reduction in such activity was observed in strains with non-specific EGSs. The inhibition of gene expression was virtually abolished at restrictive temperatures in strains that were temperature sensitive for RNase P (EC 3.1.26.5). Northern blot analysis showed that the steady-state copy number of EGS RNAs was several hundred per cell in vivo. A plasmid that contained a gene for M1 RNA covalently linked to a specific EGS reduced the level of expression of a suppressor tRNA that was encoded by a separate plasmid. Similar methods can be used to regulate gene expression in E. coli and to mimic the properties of cold-sensitive mutants. PMID- 7479949 TI - Molecular mechanism of protein-retinal coupling in bacteriorhodopsin. AB - Bacteriorhodopsin is a membrane protein that functions as a light-driven proton pump. Each cycle of proton transport is initiated by the light-induced isomerization of retinal from the all-trans to 13-cis configuration and is completed by the protein-driven reisomerization of retinal to the all-trans configuration. Previous studies have shown that replacement of Leu-93, a residue in close proximity to the 13-methyl group of retinal, by alanine, resulted in a 250-fold increase in the time required to complete each photocycle. Here, we show that the kinetic defect in the photocycle of the Leu-93-->Ala mutant occurs at a stage after the completion of proton transport and can be overcome in the presence of strong background illumination. Time-resolved retinal-extraction experiments demonstrate the continued presence of a 13-cis intermediate in the photocycle of the Leu-93-->Ala mutant well after the completion of proton release and uptake. These results indicate that retinal reisomerization is kinetically the rate-limiting step in the photocycle of this mutant and that the slow thermal reisomerization can be bypassed by the absorption of a second photon. The effects observed for the Leu-93-->Ala mutant are not observed upon replacement of any other residue in van der Waals contact with retinal or upon replacement of Leu-93 by valine. We conclude that the contact between Leu-93 and the 13-methyl group of retinal plays a key role in controlling the rate of protein conformational changes associated with retinal reisomerization and return of the protein to the initial state. PMID- 7479950 TI - Mg-SINE: a short interspersed nuclear element from the rice blast fungus, Magnaporthe grisea. AB - A short interspersed nuclear element, Mg-SINE, was isolated and characterized from the genome of the rice blast fungus, Magnaporthe grisea. Mg-SINE was isolated as an insertion element within Pot2, an inverted-repeat transposon from M. grisea and shows typical features of a mammalian SINE. Mg-SINE is present as a 0.47-kb interspersed sequence at approximately 100 copies per haploid genome in both rice and non-rice isolates of M. grisea, indicating a common evolutionary origin. Secondary structure analysis of Mg-SINE revealed a tRNA-related region at the 5' end which folds into a cloverleaf structure. Genomic fusions resulting in chimeric Mg-SINEs (Ch-SINEs) composed of a sequence homologous to Mg-SINE at the 3' end and an unrelated sequence at its 5' end were also isolated, indicating that this and other DNA rearrangements mediated by these elements may have a major effect on the genomic architecture of this fungus. PMID- 7479951 TI - Failure of programmed cell death and differentiation as causes of tumors: some simple mathematical models. AB - Most models of tumorigenesis assume that the tumor grows by increased cell division. In these models, it is generally supposed that daughter cells behave as do their parents, and cell numbers have clear potential for exponential growth. We have constructed simple mathematical models of tumorigenesis through failure of programmed cell death (PCD) or differentiation. These models do not assume that descendant cells behave as their parents do. The models predict that exponential growth in cell numbers does sometimes occur, usually when stem cells fail to die or differentiate. At other times, exponential growth does not occur: instead, the number of cells in the population reaches a new, higher equilibrium. This behavior is predicted when fully differentiated cells fail to undergo PCD. When cells of intermediate differentiation fail to die or to differentiate further, the values of growth parameters determine whether growth is exponential or leads to a new equilibrium. The predictions of the model are sensitive to small differences in growth parameters. Failure of PCD and differentiation, leading to a new equilibrium number of cells, may explain many aspects of tumor behavior--for example, early premalignant lesions such as cervical intraepithelial neoplasia, the fact that some tumors very rarely become malignant, the observation of plateaux in the growth of some solid tumors, and, finally, long lag phases of growth until mutations arise that eventually result in exponential growth. PMID- 7479952 TI - Effects of TH1 and TH2 cytokines on CD8+ cell response against human immunodeficiency virus: implications for long-term survival. AB - CD8+ cells from long-term survivors [LTS; infected with human immunodeficiency virus (HIV) for 10 or more years and having CD4+ cell counts of > or = 500 cells per microliters] have a 3-fold greater ability to suppress HIV replication than do CD8+ cells from patients who have progressed to disease (progressors) during the same time period. A change in the pattern of cytokines produced in the host from those that typically favor cell-mediated immunity (T helper 1, TH1 or type 1) to those that down-regulate it (T helper 2, TH2 or type 2) was investigated as a cause of this reduced CD8+ cell anti-HIV function. Treatment of CD8+ cells from LTS with the TH1 cytokine interleukin (IL)-2 enhanced their anti-HIV activity, whereas exposure of these cells to TH2 cytokines IL-4 or IL-10 reduced their ability to suppress HIV replication and to produce IL-2. IL-2 could prevent and reverse the inhibitory effects of IL-4 and IL-10. Moreover, prolonged exposure of CD8+ cells from some progressors to IL-2 improved the ability of these cells to suppress HIV replication. These observations support previous findings suggesting that strong CD8+ cell responses play an important role in maintaining an asymptomatic state in HIV infection. The data suggest that the loss of CD8+ cell suppression of HIV replication associated with disease progression results from a shift in cytokine production within the infected host from a TH1 to a TH2 pattern. Modulation of these cytokines could provide benefit to HIV-infected individuals by improving their CD8+ cell anti-HIV activity. PMID- 7479953 TI - An unmodified heptadecapeptide pheromone induces competence for genetic transformation in Streptococcus pneumoniae. AB - Competence for genetic transformation in Streptococcus pneumoniae has been known for three decades to arise in growing cultures at a critical cell density, in response to a secreted protease-sensitive signal. We show that strain CP1200 produces a 17-residue peptide that induces cells of the species to develop competence. The sequence of the peptide was found to be H-Glu-Met-Arg-Leu-Ser-Lys Phe-Phe-Arg-Asp-Phe-Ile-Leu-Gln-Arg- Lys-Lys-OH. A synthetic peptide of the same sequence was shown to be biologically active in small quantities and to extend the range of conditions suitable for development of competence. Cognate codons in the pneumococcal chromosome indicate that the peptide is made ribosomally. As the gene encodes a prepeptide containing the Gly-Gly consensus processing site found in peptide bacteriocins, the peptide is likely to be exported by a specialized ATP-binding cassette transport protein as is characteristic of these bacteriocins. The hypothesis is presented that this transport protein is encoded by comA, previously shown to be required for elaboration of the pneumococcal competence activator. PMID- 7479954 TI - Layer-specific programs of development in neocortical projection neurons. AB - How are long-range axonal projections from the cerebral cortex orchestrated during development? By using both passively and actively transported axonal tracers in fetal and postnatal ferrets, we have analyzed the development of projections from the cortex to a number of thalamic nuclei. We report that the projections of a cortical area to its corresponding thalamic nuclei follow highly cell-specific programs of development. Axons from cells in the deepest layers of the cerebral cortex (layer 6 and superficial subplate neurons) appear to grow very slowly and be delayed for several weeks in the cerebral white matter, reaching the thalamus over a protracted period. Neurons of layer 5, on the other hand, develop their projections much faster; despite being born after the neurons of deeper layers, layer 5 neurons are the first to extend their axons out of the cortical hemisphere and innervate the thalamus. Layer 5 projections are massive in the first postnatal weeks but may become partly eliminated later in development, being overtaken in number by layer 6 cells that constitute the major corticothalamic projection by adulthood. Layer 5 projections are area-specific from the outset and arise as collateral branches of axons directed to the brainstem and spinal cord. Our findings show that the early development of corticofugal connections is determined not by the sequence of cortical neurogenesis but by developmental programs specific for each type of projection neuron. In addition, they demonstrate that in most thalamic nuclei, layer 5 neurons (and not subplate or layer 6 neurons) establish the first descending projections from the cerebral cortex. PMID- 7479955 TI - Antibodies against the T61 antigen inhibit neuronal migration in the chick optic tectum. AB - Cell migration in the central nervous system depends, in part, on receptors and extracellular matrix molecules that likewise support axonal outgrowth. We have investigated the influence of T61, a monoclonal antibody that has been shown to inhibit growth cone motility in vitro, on neuronal migration in the developing optic tectum. Intraventricular injections of antibody-producing hybridoma cells or ascites fluid were used to determine the action of this antibody in an in vivo environment. To document alterations in tectal layer formation, a combination of cell-nuclei staining and axonal immunolabeling methods was employed. In the presence of T61 antibody, cells normally destined for superficial layers accumulated in the ventricular zone instead, leading to a reduction of the cell dense layer in the tectal plate. Experiments with 5-bromo-2'-deoxyuridine labeling followed by antibody staining confirmed that the nonmigrating cells remaining in the ventricular zone were postmitotic and had differentiated. The structure of radial glial cells, as judged by staining with a glia-specific antibody and the fluorescent tracer 1,1'-dioctadecyl-3,3,3',3' tetramethylindocarbocyanine perchlorate (DiI), remained intact in these embryos. Our findings suggest that the T61 epitope is involved in a mechanism underlying axonal extension and neuronal migration, possibly by influencing the motility of the leading process. PMID- 7479956 TI - Visual attention to surfaces in three-dimensional space. AB - Although attention plays a significant role in vision, its spatial deployment and spread in the third dimension is not well understood. In visual search experiments we show that we cannot easily focus attention across isodepth loci unless they are part of a well-formed surface with locally coplanar elements. Yet we can easily spread our attention selectively across well-formed surfaces that span an extreme range of stereoscopic depths. In cueing experiments, we show that this spread of attention is, in part, obligatory. Attentional selectivity is reduced when targets and distractors are coplanar with or rest on a common receding stereoscopic plane. We conclude that attention cannot be efficiently allocated to arbitrary depths and extents in space but is linked to and spreads automatically across perceived surfaces. PMID- 7479957 TI - Prion protein (PrP) synthetic peptides induce cellular PrP to acquire properties of the scrapie isoform. AB - Conversion of the cellular isoform of prion protein (PrPC) into the scrapie isoform (PrPSc) involves an increase in the beta-sheet content, diminished solubility, and resistance to proteolytic digestion. Transgenetic studies argue that PrPC and PrPSc form a complex during PrPSc formation; thus, synthetic PrP peptides, which mimic the conformational pluralism of PrP, were mixed with PrPC to determine whether its properties were altered. Peptides encompassing two alpha helical domains of PrP when mixed with PrPC produced a complex that displayed many properties of PrPSc. The PrPC-peptide complex formed fibrous aggregates and up to 65% of complexed PrPC sedimented at 100,000 x g for 1 h, whereas PrPC alone did not. These complexes were resistant to proteolytic digestion and displayed a high beta-sheet content. Unexpectedly, the peptide in a beta-sheet conformation did not form the complex, whereas the random coil did. Addition of 2% Sarkosyl disrupted the complex and rendered PrPC sensitive to protease digestion. While the pathogenic A117V mutation increased the efficacy of complex formation, anti PrP monoclonal antibody prevented interaction between PrPC and peptides. Our findings in concert with transgenetic investigations argue that PrPC interacts with PrPSc through a domain that contains the first two putative alpha-helices. Whether PrPC-peptide complexes possess prion infectivity as determined by bioassays remains to be established. PMID- 7479958 TI - Growth hormone secretagogues: characterization, efficacy, and minimal bioactive conformation. AB - Another class of growth hormone (GH) secretagogues has been discovered by altering the backbone structure of a flexible linear GH-releasing peptide (GHRP). In vitro and in vivo characterization confirms these GH secretagogues as the most potent and smallest (M(r) < 500) reported. Anabolic efficacy is demonstrated in rodents with intermittent delivery. A convergent model of the bioactive conformation of GHRPs is developed and is supported by the NMR structure of a highly potent cyclic analog of GHRP-2. The model and functional data provide a logical framework for the further design of low-molecular weight secretagogues and illustrate the utility of an interdisciplinary approach to elucidating potential bound-state conformations of flexible peptide ligands. PMID- 7479959 TI - The histone fold: a ubiquitous architectural motif utilized in DNA compaction and protein dimerization. AB - The histones of all eukaryotes show only a low degree of primary structure homology, but our earlier crystallographic results defined a three-dimensional structural motif, the histone fold, common to all core histones. We now examine the specific architectural patterns within the fold and analyze the nature of the amino acid residues within its functional segments. The histone fold emerges as a fundamental protein dimerization motif while the differentiations of the tips of the histone dimers appear to provide the rules of core octamer assembly and the basis for nucleosome regulation. We present evidence for the occurrence of the fold from archaebacteria to mammals and propose the use of this structural motif to define a distinct family of proteins, the histone fold superfamily. It appears that evolution has conserved the conformation of the fold even through variations in primary structure and among proteins with various functional roles. PMID- 7479960 TI - Calcium/calmodulin-dependent kinase II and long-term potentiation enhance synaptic transmission by the same mechanism. AB - Ca(2+)-sensitive kinases are thought to play a role in long-term potentiation (LTP). To test the involvement of Ca2+/calmodulin-dependent kinase II (CaM-K II), truncated, constitutively active form of this kinase was directly injected into CA1 hippocampal pyramidal cells. Inclusion of CaM-K II in the recording pipette resulted in a gradual increase in the size of excitatory postsynaptic currents (EPSCs). No change in evoked responses occurred when the pipette contained heat inactivated kinase. The effects of CaM-K II mimicked several features of LTP in that it caused a decreased incidence of synaptic failures, an increase in the size of spontaneous EPSCs, and an increase in the amplitude of responses to iontophoretically applied alpha-amino-3-hydroxy-5-methyl-4-isoxazolepropionate. To determine whether the CaM-K II-induced enhancement and LTP share a common mechanism, occlusion experiments were carried out. The enhancing action of CaM-K II was greatly diminished by prior induction of LTP. In addition, following the increase in synaptic strength by CaM-K II, tetanic stimulation failed to evoke LTP. These findings indicate that CaM-K II alone is sufficient to augment synaptic strength and that this enhancement shares the same underlying mechanism as the enhancement observed with LTP. PMID- 7479961 TI - Neural correlates of mental transformations of the body-in-space. AB - Regional cerebral blood flow was measured with positron emission tomography in human subjects during the performance of a task requiring mental rotation of their hand and a perceptually equivalent control task that did not require such a process. Comparison of the distribution of cerebral activity between these conditions demonstrated significant blood flow increases in the superior parietal cortex, the intraparietal sulcus, and the adjacent rostralmost part of the inferior parietal lobule. These findings demonstrated that, in the human brain, there is a specific system of parietal areas that are involved in mental transformations of the body-in-space. PMID- 7479962 TI - Identification of residues linked to the slow-->fast transition of thrombin. AB - Residues energetically linked to the allosteric transition of thrombin from its anticoagulant slow form to the procoagulant fast form have been identified by site-directed mutagenesis. The energetics of recognition by the two forms of the enzyme were probed by using a synthetic chromogenic substrate, fibrinogen, and hirudin. The thrombin residues E39, W60d, E192, D221, and D222 are linked to the slow-->fast transition and are part of an "allosteric core" through which events originating at the Na+ binding loop propagate to other regions of the enzyme. The thrombin residues Y76, W96, W148, and R173 lie at the periphery of the allosteric core, affect recognition of fibrinogen and hirudin to the same extent in both forms, and are not linked to the slow-->fast transition. PMID- 7479963 TI - Telomere length and replicative aging in human vascular tissues. AB - Because repeated injury of the endothelium and subsequent turnover of intimal and medial cells have been implicated in atherosclerosis, we examined telomere length, a marker of somatic cell turnover, in cells from these tissues. Telomere lengths were assessed by Southern analysis of terminal restriction fragments (TRFs) generated by HinfI/Rsa I digestion of human genomic DNA. Mean TRF length decreased as a function of population doublings in human endothelial cell cultures from umbilical veins, iliac arteries, and iliac veins. When endothelial cells were examined for mean TRF length as a function of donor age, there was a significantly greater rate of decrease for cells from iliac arteries than from iliac veins (102 bp/yr vs. 47 bp/yr, respectively, P < 0.05), consistent with higher hemodynamic stress and increased cell turnover in arteries. Moreover, the rate of telomere loss as a function of donor age was greater in the intimal DNA of iliac arteries compared to that of the internal thoracic arteries (147 bp/yr vs. 87 bp/yr, respectively, P < 0.05), a region of the arterial tree subject to less hemodynamic stress. This indicates that the effect is not tissue specific. DNA from the medial tissue of the iliac and internal thoracic arteries showed no significant difference in the rates of decrease, suggesting that chronic stress leading to cellular senescence is more pronounced in the intima than in the media. These observations extend the use of telomere size as a marker for the replicative history of cells and are consistent with a role for focal replicative senescence in cardiovascular diseases. PMID- 7479964 TI - An astrocytic binding site for neuronal Thy-1 and its effect on neurite outgrowth. AB - Thy-1, a member of the immunoglobulin superfamily, is one of the most abundant glycoproteins on mammalian neurons. Nevertheless, its role in the peripheral or central nervous system is poorly understood. Certain monoclonal antibodies to Thy 1 promote neurite outgrowth by rodent central nervous system neurons in vitro, suggesting that Thy-1 functions, in part, by modulating neurite outgrowth. We describe a binding site for Thy-1 on astrocytes. This Thy-1-binding protein has been characterized by immunofluroesence with specific anti-idiotype monoclonal antibodies and by three competitive binding assays using (i) anti-idiotype antibodies, (ii) purified Thy-1, and (iii) Thy-1-transfected cells. The Thy-1 binding protein may participate in axonal or dendritic development in the nervous system. PMID- 7479965 TI - Effects of receptor dimerization on the interaction between the class I major histocompatibility complex-related Fc receptor and IgG. AB - The neonatal Fc receptor (FcRn) transports maternal IgG from ingested milk in the gut to the bloodstream of newborn mammals. An FcRn dimer was observed in crystals of the receptor alone and of an FcRn-Fc complex, but its biological relevance was unknown. Here we use surface plasmon resonance-based biosensor assays to assess the role of FcRn dimerization in IgG binding. We find high-affinity IgG binding when FcRn is immobilized on a biosensor chip in an orientation facilitating dimerization but not when its orientation disrupts dimerization. This result supports a model in which IgG-induced dimerization of FcRn is relevant for signaling the cell to initiate endocytosis of the IgG-FcRn complex. PMID- 7479967 TI - Periodicity of strong nucleosome positioning sites around the chicken adult beta globin gene may encode regularly spaced chromatin. AB - Positioned nucleosomes contribute to both the structure and the function of the chromatin fiber and can play a decisive role in controlling gene expression. We have mapped, at high resolution, the translational positions adopted by limiting amounts of core histone octamers reconstituted onto 4.4 kb of DNA comprising the entire chicken adult beta-globin gene, its enhancer, and flanking sequences. The octamer displays extensive variation in its affinity for different positioning sites, the range exhibited being about 2 orders of magnitude greater than that of the initial binding of the octamer. Strong positioning sites are located 5' and 3' of the globin gene and in the second intron but are absent from the coding regions. These sites exhibit a periodicity (approximately 200 bp) similar to the average spacing of nucleosomes on the inactive beta-globin gene in vivo, which could indicate their involvement in packaging the gene into higher-order chromatin structure. Overlapping, alternative octamer positioning sites commonly exhibit spacings of 20 and 40 bp, but not of 10 bp. These short-range periodicities could reflect features of the core particle structure contributing to the pronounced sequence-dependent manner in which the core histone octamer interacts with DNA. PMID- 7479966 TI - Immunosuppressive drugs prevent a rapid dephosphorylation of transcription factor NFAT1 in stimulated immune cells. AB - The immunosuppressive drugs cyclosporin A and FK506 interfere with the inducible transcription of cytokine genes in T cells and in other immune cells, in part by preventing the activation of NF-AT (nuclear factor of activated T cells). We show that transcription factor NFAT1 in T cells is rapidly dephosphorylated on stimulation, that dephosphorylation occurs before translocation of NFAT1 into the cell nucleus, and that dephosphorylation increases the affinity of NFAT1 for its specific sites in DNA. Cyclosporin A prevents the dephosphorylation and the nuclear translocation of NFAT1 in T cells, B cells, macrophages, and mast cells, delineating at least one mechanism that contributes to the profound immunosuppressive effects of this compound. PMID- 7479968 TI - Hepatitis B virus HBx protein deregulates cell cycle checkpoint controls. AB - The human hepatitis B virus (HBV) HBx protein is a small transcriptional activator that is essential for virus infection. HBx is thought to be involved in viral hepatocarcinogenesis because it promotes tumorigenesis in transgenic mice. HBx activates the RAS-RAF-mitogen-activated protein (MAP) kinase signaling cascade, through which it activates transcription factors AP-1 and NF-kappa B, and stimulates cell DNA synthesis. We show that HBx stimulates cell cycle progression, shortening the emergence of cells from quiescence (G0) and entry into S phase by at least 12 h, and accelerating transit through checkpoint controls at G0/G1 and G2/M. Compared with serum stimulation, HBx was found to strongly increase the rate and level of activation of the cyclin-dependent kinases CDK2 and CDC2, and their respective active association with cyclins E and A or cyclin B. HBx is also shown to override or greatly reduce serum dependence for cell cycle activation. Both HBx and serum were found to require activation of RAS to stimulate cell cycling, but only HBx could shorten checkpoint intervals. HBx therefore stimulates cell proliferation by activating RAS and a second unknown effector, which may be related to its reported ability to induce prolonged activation of JUN or to interact with cellular p53 protein. These data suggest a molecular mechanism by which HBx likely contributes to viral carcinogenesis. By deregulating checkpoint controls, HBx could participate in the selection of cells that are genetically unstable, some of which would accumulate unrepaired transforming mutations. PMID- 7479969 TI - Membrane transport mechanisms probed by capacitance measurements with megahertz voltage clamp. AB - We have used capacitance measurements with a 1-microsecond voltage clamp technique to probe electrogenic ion-transporter interactions in giant excised membrane patches. The hydrophobic ion dipicrylamine was used to test model predictions for a simple charge-moving reaction. The voltage and frequency dependencies of the apparent dipicrylamine-induced capacitance, monitored by 1-mV sinusoidal perturbations, correspond to single charges moving across 76% of the membrane field at a rate of 9500 s-1 at 0 mV. For the cardiac Na,K pump, the combined presence of cytoplasmic ATP and sodium induces an increase of apparent membrane capacitance which requires the presence of extracellular sodium. The dependencies of capacitance changes on frequency, voltage, ATP, and sodium verify that phosphorylation enables a slow, 300- to 900-s-1, pump transition (the E1-E2 conformational change), which in turn enables fast, electrogenic, extracellular sodium binding reactions. For the GAT1 (gamma-aminobutyric acid,Na,Cl) cotransporter, expressed in Xenopus oocyte membrane, we find that chloride binding from the cytoplasmic side, and probably sodium binding from the extracellular side, results in a decrease of membrane capacitance monitored with 1- to 50-kHz perturbation frequencies. Evidently, ion binding by the GAT1 transporter suppresses an intrinsic fast charge movement which may originate from a mobility of charged residues of the transporter binding sites. The results demonstrate that fast capacitance measurements can provide new insight into electrogenic processes closely associated with ion binding by membrane transporters. PMID- 7479971 TI - Stabilization of tetanus and diphtheria toxoids against moisture-induced aggregation. AB - The progress toward single-dose vaccines has been limited by the poor solid-state stability of vaccine antigens within controlled-release polymers, such as poly(lactide-co-glycolide). For example, herein we report that lyophilized tetanus toxoid aggregates during incubation at 37 degrees C and elevated humidity -i.e., conditions relevant to its release from such systems. The mechanism and extent of this aggregation are dependent on the moisture level in the solid protein, with maximum aggregation observed at intermediate moisture contents. The main aggregation pathway is consistent with formaldehyde-mediated cross-linking, where reactive electrophiles created and stored in the vaccine upon formalinization (exposure to formaldehyde during vaccine preparation) react with nucleophiles of a second vaccine molecule to form intermolecular cross-links. This process is inhibited by the following: (i) succinylating the vaccine to block reactive amino groups; (ii) treating the vaccine with sodium cyanoborohydride, which presumably reduces Schiff bases and some other electrophiles created upon formalinization; and (iii) addition of low-molecular weight excipients, particularly sorbitol. The moisture-induced aggregation of another formalinized vaccine, diphtheria toxoid, is also retarded by succinylation, suggesting the generality of this mechanism for formalinized vaccines. Hence, mechanistic stability studies of the type described herein may be important for the development of effective single-dose vaccines. PMID- 7479972 TI - Inhibition of human immunodeficiency virus type 1 transcription and replication by DNA sequence-selective plant lignans. AB - A plant lignan, 3'-O-methyl nordihydroguaiaretic acid (3'-O-methyl NDGA, denoted Malachi 4:5-6 or Mal.4; molecular weigth 316), was isolated from Larrea tridentata and found to be able to inhibit human immunodeficiency virus (HIV) Tat regulated transactivation in vivo, induce protection of lymphoblastoid CEM-SS cells from HIV (strain IIIB) killing, and suppress the replication of five HIV-1 strains (WM, MN, VS, JR-CSF, and IIIB) in mitogen-stimulated peripheral blood mononuclear cells, all in a dose-dependent manner. Mal.4 inhibits both basal transcription and Tat-regulated transactivation in vitro. The target of Mal.4 has been localized to nucleotides -87 to -40 of the HIV long terminal repeat. Mal.4 directly and specifically interferes with the binding of Sp1 to Sp1 sites in the HIV long terminal repeat. By inhibiting proviral expression, Mal.4 may be able to interrupt the life cycles of both wild-type and reverse transcriptase or protease mutant viruses in HIV-infected patients. PMID- 7479970 TI - Th1 CD4+ lymphocytes delete activated macrophages through the Fas/APO-1 antigen pathway. AB - The Fas/APO-1 cytotoxic pathway plays an important role in the regulation of peripheral immunity. Recent evidence indicates that this regulatory function operates through deletion of activated T and B lymphocytes by CD4+ T cells expressing the Fas ligand. Because macrophages play a key role in peripheral immunity, we asked whether Fas was involved in T-cell-macrophage interactions. Two-color flow cytometry revealed that Fas receptor (FasR) was expressed on resting murine peritoneal macrophages. FasR expression was upregulated after activation of macrophages with cytokines or lipopolysaccharide, although only tumor necrosis factor-alpha rendered macrophages sensitive to anti-FasR antibody mediated death. To determine the consequence of antigen presentation by macrophages to CD4+ T cells, macrophages were pulsed with antigen and then incubated with either Th1 or Th2 cell lines or clones. Th1, but not Th2, T cells induced lysis of 60-80% of normal macrophages, whereas macrophages obtained from mice with mutations in the FasR were totally resistant to Th1-mediated cytotoxicity. Macrophage cytotoxicity depended upon specific antigen recognition by T cells and was major histocompatibility complex restricted. These findings indicate that, in addition to deletion of activated lymphocytes, Fas plays an important role in deletion of activated macrophages after antigen presentation to Th1 CD4+ T cells. Failure to delete macrophages that constitutively present self antigens may contribute to the expression of autoimmunity in mice deficient in FasR (lpr) or Fas ligand (gld). PMID- 7479973 TI - Blockade of action potential activity alters initial arborization of thalamic axons within cortical layer 4. AB - In the formation of connections during the development of the nervous system, it is generally accepted that there is an early phase not requiring neural activity and a later activity-dependent phase. The initial processes of axonal pathfinding and target selection are not thought to require neural activity, whereas the later fine-tuning of connections into their final adult patterns does. We report an apparent exception to this rule in which action potential activity seems to be required very early in development for thalamic axons to form appropriate patterns of terminal arborizations with their ultimate target neurons in layer 4 of the cerebral cortex. Blockade of sodium action potentials during the 2-week fetal period when visual thalamic axons initially grow into the primary visual cortex in cats prevents the normally occurring branching of lateral geniculate nucleus axons within layer 4. This observation implies a role for action potential activity in cerebral cortical development far earlier than previously suspected, weeks before eye-opening and the onset of the well-known process of activity-dependent reorganization of axonal terminal arbors that leads to the formation of ocular dominance columns. PMID- 7479974 TI - CD27-CD70 interactions regulate B-cell activation by T cells. AB - CD27, a member of the tumor necrosis factor (TNF) receptor family, binds to its ligand CD70, a member of the TNF family, and subsequently induces T-cell costimulation and B-cell activation. CD27 is expressed on resting T and B cells, whereas CD70 is expressed on activated T and B cells. Utilizing transfected murine pre-B-cell lines expressing human CD27 or CD70, we have examined the effect of such transfectant cells on human B-cell IgG production and B-cell proliferation. We show that the addition of CD27-transfected cells to a T-cell dependent, pokeweed mitogen-driven B-cell IgG synthesis system resulted in marked inhibition of IgG production, whereas the addition of CD70-transfected cells enhanced IgG production. The inhibition and enhancement of pokeweed mitogen driven IgG production by CD27 and CD70 transfectants were abrogated by pretreatment with anti-CD27 and anti-CD70 monoclonal antibodies, respectively. In contrast, little or no inhibition of IgG production and B-cell proliferation was noted with CD27-transfected cells or either anti-CD27 or CD70 monoclonal antibody in a T-cell-independent Staphylococcus aureus/interleukin 2-driven B-cell activation system. In this same system CD70-transfected cells enhanced B-cell IgG production and B-cell proliferation, and this enhancement could be gradually abrogated by addition of increasing numbers of CD27-transfected cells. These results clearly demonstrate that interactions among subsets of T cells expressing CD27 and CD70 play a key role in regulating B-cell activation and immunoglobulin synthesis. PMID- 7479975 TI - Fluidity of the lipid domain of cell wall from Mycobacterium chelonae. AB - The mycobacterial cell wall contains large amounts of unusual lipids, including mycolic acids that are covalently linked to the underlying arabinogalactan peptidoglycan complex. Hydrocarbon chains of much of these lipids have been shown to be packed in a direction perpendicular to the plane of the cell surface. In this study, we examined the dynamic properties of the organized lipid domains in the cell wall isolated from Mycobacterium chelonae grown at 30 degrees C. Differential scanning calorimetry showed that much of the lipids underwent major thermal transitions between 30 degree C and 65 degrees C, that is at temperatures above the growth temperature, a result suggesting that a significant portion of the lipids existed in a structure of extremely low fluidity in the growing cells. Spin-labeled fatty acid probes were successfully inserted into the more fluid part of the cell wall. Our model of the cell wall suggests that this domain corresponds to the outermost leaflet, a conclusion reinforced by the observation that labeling of intact cells produced electron spin resonance spectra similar to those of the isolated cell wall. Use of stearate labeled at different positions showed that the fluidity within the outer leaflet increased only slightly as the nitroxide group was placed farther away from the surface. These results are consistent with the model of mycobacterial cell wall containing an asymmetric lipid bilayer, with an internal, less fluid mycolic acid leaflet and an external, more fluid leaflet composed of lipids containing shorter chain fatty acids. The presence of the low-fluidity layer will lower the permeability of the cell wall to lipophilic antibiotics and chemotherapeutic agents and may contribute to the well-known intrinsic resistance of mycobacteria to such compounds. PMID- 7479976 TI - Signal-induced degradation of I kappa B alpha requires site-specific ubiquitination. AB - The inhibitor protein I kappa B alpha controls the nuclear import of the transcription factor NF-kappa B. The inhibitory activity of I kappa B alpha is regulated from the cytoplasmic compartment by signal-induced proteolysis. Previous studies have shown that signal-dependent phosphorylation of serine residues 32 and 36 targets I kappa B alpha to the ubiquitin-proteasome pathway. Here we provide evidence that lysine residues 21 and 22 serve as the primary sites for signal-induced ubiquitination of I kappa B alpha. Conservative Lys- >Arg substitutions at both Lys-21 and Lys-22 produce dominant-negative mutants of I kappa B alpha in vivo. These constitutive inhibitors are appropriately phosphorylated but fail to release NF-kappa B in response to multiple inducers, including viral proteins, cytokines, and agents that mimic antigenic stimulation through the T-cell receptor. Moreover, these Lys-->Arg mutations prevent signal dependent degradation of I kappa B alpha in vivo and ubiquitin conjugation in vitro. We conclude that site-specific ubiquitination of phosphorylated I kappa B alpha at Lys-21 and/or Lys-22 is an obligatory step in the activation of NF-kappa B. PMID- 7479978 TI - Covalent protein-DNA complexes at the 5' strand termini of meiosis-specific double-strand breaks in yeast. AB - During meiosis in Saccharomyces cerevisiae, the first chemical step in homologous recombination is the occurrence of site-specific DNA double-strand breaks (DSBs). In wild-type cells, these breaks undergo resection of their 5' strand termini to yield molecules with 3' single-stranded tails. We have further characterized the breaks that accumulate in rad50S mutant stains defective in DSB resection. We find that these DSBs are tightly associated with protein via what appears to be a covalent linkage. When genomic DNA is prepared from meiotic rad50S cultures without protease treatment steps, the restriction fragments diagnostic of DSBs selectively partition to the organic-aqueous interphase in phenol extractions and band at lower than normal density in CsCl density gradients. Selective partitioning and decreased buoyant density are abolished if the DNA is treated with proteinase K prior to analysis. Similar results are obtained with sae2-1 mutant strains, which have phenotypes identical to rad50S mutants. The protein is bound specifically to the 5' strand termini of DSBs and is present at both 5' ends in at least a fraction of breaks. The stability of the complex to various protein denaturants and the strand specificity of the attachment are most consistent with a covalent linkage to DSB termini. We propose that the DSB associated protein is the catalytic subunit of the meiotic recombination initiation nuclease and that it cleaves DNA via a covalent protein-DNA intermediate. PMID- 7479977 TI - Cell cycle regulation of the cyclin A gene promoter is mediated by a variant E2F site. AB - Cyclin A is involved in the control of S phase and mitosis in mammalian cells. Expression of the cyclin A gene in nontransformed cells is characterized by repression of its promoter during the G1 phase of the cell cycle and its induction at S-phase entry. We show that this mode of regulation is mediated by the transcription factor E2F, which binds to a specific site in the cyclin A promoter. It differs from the prototype E2F site in nucleotide sequence and protein binding; it is bound by E2F complexes containing cyclin E and p107 but not pRB. Ectopic expression of cyclin D1 triggers premature activation of the cyclin A promoter by E2F, and this effect is blocked by the tumor suppressor protein p16INK4. PMID- 7479979 TI - Neurotrophin 3 rescues neuronal precursors from apoptosis and promotes neuronal differentiation in the embryonic metanephric kidney. AB - We analyzed the developmental regulation and role of the neurotrophins during metanephric kidney morphogenesis. RNase protection assay revealed the presence of nerve growth factor, neurotrophin 3 (NT-3), and brain-derived neurotrophic factor mRNAs and the regulation of their expression during embryonic development of rat metanephros. NT-3 induced differentiation (neurite outgrowth) and survival (inhibition of apoptosis) of the neuronal precursors in cultured nephrogenic mesenchymes and neuronal differentiation in cultured whole kidneys, whereas NT 4/5, brain-derived neurotrophic factor, and nerve growth factor were without effect. The neurotrophins did not trigger tubular differentiation of isolated nephrogenic cells, which underwent apoptosis when cultured with or without the neurotrophins. NT-3 is thus an inducer of differentiation and a survival factor for renal neuronal cells, but none of the neurotrophins is a morphogen in kidney tubule induction. PMID- 7479980 TI - A null mutation in the gene encoding a type I interferon receptor component eliminates antiproliferative and antiviral responses to interferons alpha and beta and alters macrophage responses. AB - To examine the in vivo role(s) of type I interferons (IFNs) and to determine the role of a component of the type I IFN receptor (IFNAR1) in mediating responses to these IFNs, we generated mice with a null mutation (-/-) in the IFNAR1 gene. Despite compelling evidence for modulation of cell proliferation and differentiation by type I IFNs, there were no gross signs of abnormal fetal development or morphological changes in adult IFNAR1-/- mice. However, abnormalities of hemopoietic cells were detected in IFNAR1 -/- mice. Elevated levels of myeloid lineage cells were detected in peripheral blood and bone marrow by staining with Mac-1 and Gr-1 antibodies. Furthermore, bone marrow macrophages from IFNAR1 -/- mice showed abnormal responses to colony-stimulating factor 1 and lipopolysaccharide. IFNAR1 -/- mice were highly susceptible to viral infection: viral titers were undetected 24 hr after infection of IFNAR1 +/+ mice but were extremely high in organs of IFNAR1 -/- mice, demonstrating that the type I IFN system is a major acute antiviral defence. In cell lines derived from IFNAR1 -/- mice, there was no signaling in response to IFN-alpha or -beta as measured by induction of 2'-5' oligoadenylate synthetase, antiviral, or antiproliferative responses. Importantly, these studies demonstrate that type I IFNs function in the development and responses of myeloid lineage cells, particularly macrophages, and that the IFNAR1 receptor component is essential for antiproliferative and antiviral responses to IFN-alpha and -beta. PMID- 7479981 TI - Invariant chain made in Escherichia coli has an exposed N-terminal segment that blocks antigen binding to HLA-DR1 and a trimeric C-terminal segment that binds empty HLA-DR1. AB - Invariant chain (Ii), a membrane glycoprotein, binds class II major histocompatibility complex (MHC) glycoproteins, probably via its class II associated Ii peptide (CLIP) segment, and escorts them toward antigen-containing endosomal compartments. We find that a soluble, trimeric ectodomain of Ii expressed and purified from Escherichia coli blocks peptide binding to soluble HLA-DR1. Proteolysis indicates that Ii contains two structural domains. The C terminal two-thirds forms an alpha-helical domain that trimerizes and interacts with empty HLA-DR1 molecules, augmenting rather than blocking peptide binding. The N-terminal one-third, which inhibits peptide binding, is proteolytically susceptible over its entire length. In the trimer, the N-terminal domains act independently with each CLIP segment exposed and free to bind an MHC class II molecule, while the C-terminal domains act as a trimeric unit. PMID- 7479982 TI - Spontaneous inflammatory demyelinating disease in transgenic mice showing central nervous system-specific expression of tumor necrosis factor alpha. AB - Cytokines are now recognized to play important roles in the physiology of the central nervous system (CNS) during health and disease. Tumor necrosis factor alpha (TNF-alpha) has been implicated in the pathogenesis of several human CNS disorders including multiple sclerosis, AIDS dementia, and cerebral malaria. We have generated transgenic mice that constitutively express a murine TNF-alpha transgene, under the control of its own promoter, specifically in their CNS and that spontaneously develop a chronic inflammatory demyelinating disease with 100% penetrance from around 3-8 weeks of age. High-level expression of the transgene was seen in neurons distributed throughout the brain. Disease is manifested by ataxia, seizures, and paresis and leads to early death. Histopathological analysis revealed infiltration of the meninges and CNS parenchyma by CD4+ and CD8+ T lymphocytes, widespread reactive astrocytosis and microgliosis, and focal demyelination. The direct action of TNF-alpha in the pathogenesis of this disease was confirmed by peripheral administration of a neutralizing anti-murine TNF alpha antibody. This treatment completely prevented the development of neurological symptoms, T-cell infiltration into the CNS parenchyma, astrocytosis, and demyelination, and greatly reduced the severity of reactive microgliosis. These results demonstrate that overexpression of TNF-alpha in the CNS can cause abnormalities in nervous system structure and function. The disease induced in TNF-alpha transgenic mice shows clinical and histopathological features characteristic of inflammatory demyelinating CNS disorders in humans, and these mice represent a relevant in vivo model for their further study. PMID- 7479983 TI - Nitric oxide inhibits the release of norepinephrine and dopamine from the medial basal hypothalamus of the rat. AB - Previous research indicates that norepinephrine and dopamine stimulate release of luteinizing hormone (LH)-releasing hormone (LHRH), which then reaches the adenohypophysis via the hypophyseal portal vessels to release LH. Norepinephrine exerts its effect via alpha 1-adrenergic receptors, which stimulate the release of nitric oxide (NO) from nitricoxidergic (NOergic) neurons in the medial basal hypothalamus (MBH). The NO activates guanylate cyclase and cyclooxygenase, thereby inducing release of LHRH into the hypophyseal portal vessels. We tested the hypothesis that these two catecholamines modulate NO release by local feedback. MBH explants were incubated in the presence of sodium nitroprusside (NP), a releaser of NO, and the effect on release of catecholamines was determined. NP inhibited release of norepinephrine. Basal release was increased by incubation of the tissue with the NO scavenger hemoglobin (20 micrograms/ml). Hemoglobin also blocked the inhibitory effect of NP. In the presence of high potassium (40 mM) medium to depolarize cell membranes, norepinephrine release was increased by a factor of 3, and this was significantly inhibited by NP. Hemoglobin again produced a further increase in norepinephrine release and also blocked the action of NP. When constitutive NO synthase was inhibited by the competitive inhibitor NG-monomethyl-L-arginine (NMMA) at 300 microM, basal release of norepinephrine was increased, as was potassium-evoked release, and this was associated in the latter instance with a decrease in tissue concentration, presumably because synthesis did not keep up with the increased release in the presence of NMMA. The results were very similar with dopamine, except that reduction of potassium-evoked dopamine release by NP was not significant. However, the increase following incubation with hemoglobin was significant, and hemoglobin, when incubated with NP, caused a significant elevation in dopamine release above that with NP alone. In this case, NP increased tissue concentration of dopamine along with inhibiting release, suggesting that synthesis continued, thereby raising the tissue concentration in the face of diminished release. When the tissue was incubated with NP plus hemoglobin, which caused an increase in release above that obtained with NP alone, the tissue concentration decreased significantly compared with that in the absence of hemoglobin, indicating that, with increased release, release exceeded synthesis, causing a fall in tissue concentration. When NO synthase was blocked by NMMA, the release of dopamine, under either basal or potassium-evoked conditions, was increased. Again, in the latter instance the tissue concentration declined significantly, presumably because synthesis did not match release. Therefore, the results were very similar with both catecholamines and indicate that NO acts to suppress release of both amines. Since both catecholamines activate the release of LHRH, the inhibition of their release by NO serves as an ultra-short-loop negative feedback by which NO inhibits the release of the catecholamines, thereby reducing the activation of the NOergic neurons and decreasing the release of LHRH. This may be an important means for terminating the pulses of release of LHRH, which generate the pulsatile release of LH that stimulates gonadal function in both male and female mammals. PMID- 7479984 TI - Spatial heterogeneity of inhibitory surrounds in the middle temporal visual area. AB - A recurrent theme in the organization of vertebrate visual cortex is that of receptive fields with an associated "silent" opponency component. In the middle temporal area (area MT), a cortical visual area involved in the analysis of retinal motion in primates, this opponency appears in the form of a region outside the classical receptive field (CRF) that in itself gives no response but suppresses responses to motion evoked within the CRF. This antagonistic motion surround has been described as very large and symmetrically arrayed around the CRF. On the basis of this view, the primary function of the surround has long been thought to consist of simple figure-ground segregation based on movement. We have made use of small stimulus patches to map the form and extent of the surround and find evidence that the surround inhibition of many MT cells is in fact confined to restricted regions on one side or on opposite sides of the CRF. Such regions endow MT cells with the ability to make local-to-local motion comparisons, capable of extracting more complex features from the visual environment, and as such, may be better viewed as intrinsic parts of the receptive field, rather than as separate entities responsible for local-to-global comparisons. PMID- 7479985 TI - Evidence for synaptotagmin as an inhibitory clamp on synaptic vesicle release in Aplysia neurons. AB - While previous studies have demonstrated that synaptotagmin plays an essential role in evoked neurotransmitter release, it has been difficult to determine whether it acts to facilitate or inhibit release. To address this question, we used acute genetic manipulations to alter the expression of synaptotagmin in Aplysia neurons. Transient overexpression of synaptotagmin in acutely dissected cholinergic neurons and in cultured glutaminergic neurons decreased the amplitude of the excitatory postsynaptic potential (EPSP) by 32% and 26%, respectively. In contrast, treatment of cultured presynaptic neurons with synaptotagmin antisense oligonucleotides increased the amplitude of the EPSP by 50-75%. These results are consistent with a role of synaptotagmin as an inhibitor of release. PMID- 7479986 TI - Inhibition of ascorbate peroxidase by salicylic acid and 2,6-dichloroisonicotinic acid, two inducers of plant defense responses. AB - In recent years, it has become apparent that salicylic acid (SA) plays an important role in plant defense responses to pathogen attack. Previous studies have suggested that one of SA's mechanisms of action is the inhibition of catalase, resulting in elevated levels of H2O2, which activate defense-related genes. Here we demonstrate that SA also inhibits ascorbate peroxoidase (APX), the other key enzyme for scavenging H2O2. The synthetic inducer of defense responses, 2,6-dichloroisonicotinic acid (INA), was also found to be an effective inhibitor of APX. In the presence of 750 microM ascorbic acid (AsA), substrate-dependent IC50 values of 78 microM and 95 microM were obtained for SA and INA, respectively. Furthermore, the ability of SA analogues to block APX activity correlated with their ability to induce defense-related genes in tobacco and enhance resistance to tobacco mosaic virus. Inhibition of APX by SA appears to be reversible, thus differing from the time-dependent, irreversible inactivation by suicide substrates such as p-aminophenol. In contrast to APX, the guaiacol utilizing peroxidases, which participate in the synthesis and crosslinking of cell wall components as part of the defense response, are not inhibited by SA or INA. The inhibition of both catalase and APX, but not guaiacol peroxidases, supports the hypothesis that SA-induced defense responses are mediated, in part, through elevated H2O2 levels or coupled perturbations of the cellular redox state. PMID- 7479987 TI - Inferring phylogenies from DNA sequences of unequal base compositions. AB - A new method for computing evolutionary distances between DNA sequences is proposed. Contrasting with classical methods, the underlying model does not assume that sequence base compositions (A, C, G, and T contents) are at equilibrium, thus allowing unequal base compositions among compared sequences. This makes the method more efficient than the usual ones in recovering phylogenetic trees from sequence data when base composition is heterogeneous within the data set, as we show by using both simulated and empirical data. When applied to small-subunit ribosomal RNA sequences from several prokaryotic or eukaryotic organisms, this method provides evidence for an early divergence of the microsporidian Vairimorpha necatrix in the eukaryotic lineage. PMID- 7479988 TI - Boldness and predator deterrence: a critique of Godin & Davis. AB - Godin & Davis (1995) claimed to have provided experimental evidence for a predator pursuit deterrence function of predator inspection in the guppy. We doubt that they have shown that inspection behaviour per se deters attack because their evidence is actually correlational and thus inherently ambiguous; furthermore we think their methods are flawed. PMID- 7479990 TI - The Wellcome Foundation Lecture, 1994. The fetal origins of adult disease. AB - Recent findings suggest that many human fetuses have to adapt to a limited supply of nutrients and in doing so they permanently change their physiology and metabolism. These 'programmed' changes may be the origins of a number of diseases in later life, including coronary heart disease and the related disorders: stroke, diabetes and hypertension. PMID- 7479989 TI - Comparison of a mitochondrial gene and a corresponding nuclear pseudogene. AB - Nuclear copies of mitochondrial genes have been reported several times. Presented here is a direct comparison of a fragment of the mitochondrial gene coding for Cytochrome b and its assumed nuclear pseudogene in a phylogenetic context. By studying eight such sets of genes a direct measurement of relative rates of several types of substitutions were made. As expected mitochondrial third position transitions are the fastest accumulating substitutions, here indicated to be at least up to 39 times faster than corresponding positions in the supposed nuclear pseudogene. Translocated mitochondrial genes, evolving much slower than their functional 'counterpart', reflect the ancestral-pre-translocated form of the gene. A warning is given against unwanted inclusion of paralogous sequences in phylogenetic analysis and against the use of versatile primers that can promote such incidents. PMID- 7479991 TI - Delayed density-dependence in a small-rodent population. AB - The role of delayed density-dependent processes in the dynamics of animal populations poses a problem for ecologists; although generally assumed important in populations that show cyclic or chaotic fluctuations, little experimental evidence for such processes exist. Through manipulation of vole densities within enclosed areas it was shown that reproduction, recruitment, and body growth rate in introduced populations were negatively affected by high previous density. In addition, female movement patterns shifted, and territoriality as well as home range size was increased after high density. The observed changes in female spacing-behaviour suggested that negative effects of previous density were partly mediated by social interactions, and agreed with the finding that smaller (less competitive) females were the ones suffering most from increased competition. Contrary to expectations from recent work, predation could be excluded as the cause of delayed density-dependence in this study. Instead, chemical analyses of a dominating food plant suggested that herbivory at high vole-density had delayed negative effects on food quality. PMID- 7479992 TI - Calibration of thromboplastins. AB - Anticoagulant therapy with coumarin drugs requires tests on the patient's blood to determine the dose rate for the drug. The tests involve measuring the effect of a preparation of a certain reagent, thromboplastin, in accelerating the clotting of the patient's blood. Features of the World Health Organization's recommendation on the appropriate laboratory procedure for a calibration assay and evaluation of results call for comment. The standard method involves estimation of a linear structural relation, a statistical technique long known but seldom finding practical application. This paper reviews its history, as a prelude to discussing strange mistakes in earlier publications; it also examines assessment of the precision of estimation and other aspects of the statistical method. PMID- 7479993 TI - The variance of successive peaks in synaptic amplitude histograms: effects of inter-site differences in quantal size. AB - Variability in the measured amplitude of evoked synaptic events can arise from several factors, including: measurement noise, trial-to-trial variation in the amplitude of the response at a single release site, or variation between different release sites (inter-site variation) in the mean amplitude of the single quantal response. Classic (linear) models of variability include only the first two of these factors, although differences in the number of postsynaptic receptors or the degree of electrotonic attenuation for different release sites could cause substantial inter-site variations in quantal size. In this paper, the effect of inter-site variation on the variance of successive histogram peaks has been determined analytically and verified by computational studies. This effect is minimal at the edges of the histogram and contributes maximally to central peaks. Linear approximations to the variance of successive histogram peaks may therefore result in very poor fits to measured data if substantial inter-site variation in quantal size is involved. Our computational results indicate that for synaptic contacts with high release probabilities and substantial inter-site variation, the variance of histogram peaks will decrease with increasing quantal content. PMID- 7479994 TI - Multiple origins of anaerobic ciliates with hydrogenosomes within the radiation of aerobic ciliates. AB - Some ciliates live anaerobically and lack mitochondria, but possess hydrogenosomes: organelles that contain hydrogenase and produce hydrogen. The origin of hydrogenosomes has been explained by two competing hypotheses: (i) they are biochemically modified mitochondria; or (ii) they are derived from endosymbiotic association(s) of ciliates and anaerobic eubacteria that possessed the hydrogenosome biochemistry. Phylogenetic analyses of representative aerobic, and anaerobic hydrogenosomal ciliates using host nuclear SSU rDNA sequences indicate a minimum of three, but more likely four, separate origins of hydrogenosomes. Whereas this does not refute either hypothesis, the implausibility of multiple convergent endosymbioses gives further support to the view that hydrogenosomes in ciliates derive from an existing organelle, which ultrastructural evidence suggests is the mitochondrion. Our results indicate a considerable potential for physiological-biochemical plasticity among a group of predominantly aerobic eucaryotes, and provide a phylogenetic framework to further refine and test hypotheses of the origins of the hydrogenosomal enzymes. PMID- 7479996 TI - Cover illustration--regional sources of natural products: Taxomyces andreanae. PMID- 7479995 TI - Development and migration of olfactory neurones in the nervous system of the neonatal opossum. AB - The neonatal opossum (Monodelphis domestica) was used to assess how different populations of cells are generated in the olfactory region, and how they migrate along pathways to the central nervous system. Developing nerve cells were immunocytochemically labelled using antisera directed against two specific markers of olfactory receptor neurones: olfactory marker protein (OMP) and the dipeptide carnosine. In new-born opossums both carnosine and OMP are already co expressed in primary olfactory neurones and in those axons that extend towards the olfactory bulb. Expression of these markers in olfactory receptor neurones during the first postnatal days reflects the advanced developmental state of this system compared to other regions of the central nervous system (such as the cortex and cerebellum), which are highly immature and less developed in comparison with those of new-born rats or mice. A second, distinct population of carnosine/OMP expressing cells was also identified during the first postnatal week. These neurones were present as clusters along the olfactory nerve bundles, on the ventral-medial aspect of the olfactory bulb and in the basal prosencephalon. The distribution of this cell population was compared to another group of well characterized migratory neurones derived from the olfactory placode, which express the decapeptide GnRH (Gonadotropin-releasing hormone, also known as LHRH). GnRH was never co-localized with carnosine/OMP in the same migratory cells. These observations show that distinct cell populations arise from the olfactory placode in the neonatal opossum and that they migrate to colonize the central nervous system by following common pathways. PMID- 7479997 TI - Melatonin receptors potentiate contractile responses to adrenergic nerve stimulation in rat caudal artery. PMID- 7479998 TI - A comparison of the action of mexiletine and morphine in rodent models of acute and chronic pain. PMID- 7479999 TI - Effects of chaparrin, nordihydroguaiaretic acid and their structural analogues on Entamoeba histolytica cultures. PMID- 7480000 TI - Development-related changes in colon muscarinic receptors. PMID- 7480001 TI - Lead-induced alteration in vascular function of rat aorta. PMID- 7480002 TI - A model for beta-amyloid aggregation and neurotoxicity based on the free radical generating capacity of the peptide: implications of "molecular shrapnel" for Alzheimer's disease. PMID- 7480003 TI - Alpha 1-adrenoceptors: pharmacological classification and newer therapeutic applications. PMID- 7480004 TI - Cholinergic channel activators: novel opportunities for the treatment of CNS disorders. AB - Negative connotations associated with the use of (-)-nicotine has limited medicinal chemistry research in the area of nAChRs [5]. However, recent evidence suggests that a diversity of nAChR subunits exist, that each subtype may be involved in mediating specific neurochemical/behaviors, and that these subtypes have a defined pharmacology that may be selectively targeted [1]. (+/-) Epibatidine, GTS-21 and ABT-418 differentially interact with nAChR subtypes to elicit a diversity of behavioral effects including analgesia, neuroprotection and cognitive enhancement. These agents therefore represent important new pharmacological probes to dissect the nAChR subtype(s) mediating specific pharmacological responses to nAChR activation. PMID- 7480005 TI - Effects of long-term treatment with a monoamine oxidase B inhibitor, selegeline (L-deprenyl), on amino acid transmitter systems in the ageing rat brain. PMID- 7480006 TI - Hyaluronan receptor antagonists alter skin inflammation and fibrosis following injury. PMID- 7480007 TI - Drug discovery and development in academic institutions. PMID- 7480009 TI - Nitrovasodilator-induced relaxation in guinea pig vascular and tracheal smooth muscle. PMID- 7480008 TI - Efficacy of peptide and nonpeptidic agonists at the cloned human delta opioid receptor. PMID- 7480010 TI - Endotoxin-induced hypotension exhibits seasonal variation: a role for the submandibular glands? PMID- 7480011 TI - Do related kappa agonists produce similar effects on cardiac ion channels? PMID- 7480012 TI - all-trans-retinol modulation of nitronaphthalene-induced lung and liver injury in male Sprague-Dawley rats. PMID- 7480013 TI - Nitric oxide synthase antagonism potentiates pressure-flow autoregulation in the superior mesenteric artery. PMID- 7480014 TI - A selective Ascaris acetylcholine receptor antagonist: 1,1-dimethyl-4-(2 quinolyl)-piperazinium iodide. PMID- 7480015 TI - Effects of the pyrrolizidine alkaloid, retrorsine, on sulfur metabolism, in the liver. PMID- 7480016 TI - Similarity of effects of N8-acetylspermidine and its deacetylase inhibitor on stimulating the growth of L1210 cells. PMID- 7480017 TI - Effects of barbital or diazepam withdrawal on seizure thresholds to bicuculline, picrotoxin and flurazepam. PMID- 7480018 TI - In vitro cytotoxicity of chaparrin from Castela texana. PMID- 7480020 TI - Serotoninergic 5-HT1-like receptors of the rabbit ear artery: how silent are they? PMID- 7480019 TI - Analeptic effect of TRH in rats selectively bred for differential CNS sensitivity to ethanol. PMID- 7480021 TI - Immobilization of frog sperm by exposure to Ruta chalepensis extract. PMID- 7480022 TI - Determination of the cyclic GMP concentration yielding half-maximal activation of cyclic GMP-dependent protein kinase in intact ovine basilar arteries. PMID- 7480023 TI - Metabolic activation of carcinogenic arylamines by rapid acetylator, slow acetylator, and chimeric recombinant Syrian hamster NAT2 allozymes. PMID- 7480024 TI - Interleukin-1 beta induces nerve growth factor release and gene expression in human glioblastoma cells. PMID- 7480025 TI - Pregnancy-related alterations of vascular responses in rat. PMID- 7480026 TI - The dermal toxicity of industrial metal-working lubricants. PMID- 7480028 TI - Partial left ventricular ischemia in isolated hearts obtained from ethanol dependent rats. PMID- 7480029 TI - Effects of imipramine on the blood pressure of pithed rats. PMID- 7480027 TI - Clinical testing of "amole" (Agave lecheguilla): phase 1-A. PMID- 7480030 TI - Differential effects of histamine on T helper type 2 (TH2) lymphocytes. AB - Histamine inhibits IL-4 secretion from T helper type 2 cells but enhances IL-5 secretion from the same cells. Both of these effects are mediated by H2 receptors. Not all cyclic AMP-elevating agents affect IL-4 and IL-5 secretion. The observations suggest a selective effect of histamine on TH2 lymphocytes. Furthermore, Cl- channels may be involved in histamine-mediated regulation of cytokine secretion. The data suggest that histamine-induced elevation of cyclicAMP may be an important, but probably not the sole, mechanism underlying the selective effects of histamine on cytokine production. PMID- 7480031 TI - M1 muscarinic receptor blockade causes insulin resistance in the cat. PMID- 7480032 TI - The activation of serotonergic 5-HT1A presynaptic receptors or an enhancement of 5-HT postsynaptic activity increase learning. PMID- 7480033 TI - Effect of serum lipoprotein fractions from hypertriglyceridemic patients upon some rat liver mitochondrial functions. PMID- 7480034 TI - Interaction of glutathione derivatives with brain 2-amino-3-hydroxy-5-methyl-4 isoxazole propionate (AMPA) receptors. PMID- 7480035 TI - Direct vascular effects of thiopental. PMID- 7480037 TI - Effects of chronic consumption of a salted diet and exogenous administration of capsaicin on the gastric mucosa of the rat during postnatal development. PMID- 7480036 TI - Contractile actions of thrombin receptor-derived polypeptides in rat and guinea pig lung parenchymal smooth muscle. PMID- 7480038 TI - Intracellular calcium buffering in adrenergic nerves: evidence for decline in the aged rat. PMID- 7480039 TI - [Management and the quality of nursing care]. PMID- 7480040 TI - [Rehabilitation and education of the patient with Down syndrome]. PMID- 7480042 TI - [The price of certain beliefs]. PMID- 7480041 TI - [The fight against poliomyelitis. Search for a vaccine]. PMID- 7480043 TI - [Europe against Cancer. A project of the Permanent Committee of Nurses for basic nursing education]. PMID- 7480044 TI - [Psychosocial aspects of burnout]. PMID- 7480045 TI - [The hospital discharge card. Problems of recording diagnoses, interventions and and hospital procedures]. PMID- 7480050 TI - Improving patient moving and handling skills. AB - Training in manual handling should incorporate a problem-solving approach. The handling and moving of patients is a learned skill that needs regular revision. Assessment of patient capabilities is essential, as is the provision of a safe, ergonomically satisfactory environment. There is no single technique that works in all situations. PMID- 7480047 TI - [The nurse's relationship with the patient, his family and with the health care staff]. PMID- 7480048 TI - [Overcoming the obstacles that interfere with nursing research]. PMID- 7480051 TI - Lyme disease. PMID- 7480052 TI - Evaluating a patient teaching programme. AB - Patient teaching is an essential element of care that should involve the multidisciplinary team. Patients and their significant others must have access to patient teaching programmes in order that they may make informed choices. The design of the programme must be considered carefully in order to meet the needs of patients and their significant others. Patients clearly benefit from well organised and appropriately planned patient teaching programmes. PMID- 7480049 TI - Using a personal profile to claim CATS points. AB - Career progression in nursing increasingly requires evidence of academic as well as practical abilities. CATS claims are awarded for proof of prior learning. Universities differ in the tariffs they award for nursing courses so it pays to shop around. Developing appropriate academic study skills is more important than claiming maximum CATS points. PMID- 7480046 TI - [The system of diagnosis related groups. Classification of hospitalization episodes for the reimbursement of hospital stay]. PMID- 7480054 TI - The use of compression to treat lymphoedema. AB - Lymphoedema in breast cancer patients can be treated. Patients need to feel that they have control over arm swelling. The treatment needs to be initiated by a physiotherapist working with a nurse specialist. PMID- 7480053 TI - Training staff to provide a continence helpline. AB - Incontinence is still a taboo subject with many people suffering in silence. A telephone helpline can encourage people to make contact with health professionals, but those running the helpline must be appropriately trained. Essential ingredients for a training session include assessment of prior knowledge, a clear overall aim and learning objectives, an environment condusive to learning, involvement of the learners and feedback from them. PMID- 7480055 TI - Peak flow meters. AB - Peak flow is an objective measure of respiratory status. Nurses working with people with asthma require specialist knowledge. Nurses need to have an awareness of products available for people with asthma. Self-management plans can benefit people with asthma. PMID- 7480056 TI - Defining the concept of care in the community. AB - Care in the community has meant many different policies at different times. Recent policies have changed definitions but have done little to improve the lot of people affected by mental illness. PMID- 7480057 TI - From angels to villains. PMID- 7480059 TI - Helping relatives cope with the grieving process. AB - Grief may be a natural reaction to loss, felt by those most affected, within their own cultural context. Stage-theory approaches to grief may encourage people to think of grief in a simplistic or inflexible way. It is important to allow patients to make their own responses and to express their feelings. PMID- 7480058 TI - The history of industrial action in nursing. AB - The long tradition of successful campaigning and industrial action by nurses counters the profession's complacent image. Throughout the century, nurses have fought for, and gained, improved working conditions and pay in line with inflation. The threat of local pay bargaining means that nurses continue to need the protection of their unions. PMID- 7480060 TI - Commode design for frail and disabled people. AB - Commode use carries a significant risk of falls for frail older people. Traditional commode design is often inadequate. Working with manufacturers can bring essential design changes. PMID- 7480061 TI - The psychological impact of warts on patients' lives. AB - Warts were for many years associated with witches and magic rituals. Caused by the human papilloma virus, warts are a common dermatological problem. Warts can have devastating psychological effects on patients. It is vital that patients are offered support and advice as well as treatment. PMID- 7480062 TI - Structure and function of phospholipase A2 receptor. PMID- 7480063 TI - The regulation of stearoyl-CoA desaturase (SCD). AB - The stearoyl-CoA desaturase gene family encodes stearoyl-CoA desaturase, the key enzyme involved in the biosynthesis of unsaturated fatty acids, as well as in the regulation of this process. Because of the important role that the SCD gene product plays in fat cell metabolism, future studies on SCD1 gene expression could provide new insights into the role of fatty acids in cellular regulation, metabolism, and gene expression both in normal and disease states. In addition, the SCD gene family can be used as a model to study mechanisms of cellular differentiation, tissue-specific gene expression, and dietary and hormonal regulation of gene expression. PMID- 7480064 TI - A family of phospholipid autacoids: occurrence, metabolism and bioactions. PMID- 7480065 TI - Intestinal lipoprotein synthesis and secretion. PMID- 7480066 TI - Oxylanosterols as modifiers of cholesterol biosynthesis. AB - In this review, the thinking and strategy that lead to the design of mechanism based inhibitors of cholesterol biosynthesis have been recounted. This work began with a purely biochemical perspective on the mechanism of lanosterol demethylation. The final efforts focused on pharmacology and drug design thus bringing the basic science effort to a practical application. Most recently, a series of 15-oxalanosterols, which act as pure suppressors of HMG-CoA reductase lacking lanosterol demethylase inhibition properties has been identified. These molecules also lower serum cholesterol and show promise as potential agents for clinical evaluation. The utility of these compounds and validation of our hypothesis will have to await further testing. PMID- 7480067 TI - Optimising the radioimmunotherapy of malignant disease: the broadening choice of carrier and effector moieties. AB - The treatment of cancer by radioimmunotherapy remains an experimental approach successful only in a limited number of selected disease conditions. One ground for optimism over the future of radioimmunotherapy lies in the fact that where cures have been obtained it has been despite the design of the immunoconjugate rather than because of it. As the choice of available functional components for conjugate construction increases, the process of evaluation and optimisation is underway. The replacement of the commonly used 131I with radionuclides possessing radiation characteristics more suited to particular disease states, and tailored to the behaviour of the carrier vehicle, should bring improved energy deposition within tumour while reducing whole body radiation burden. Similarly, the introduction of chemically or genetically engineered targeting molecules in place of conventional antibodies may bring improved pharmacokinetic characteristics and higher tumour accumulation. Optimisation of the therapeutic and carrier moieties employed in radioimmunotherapy should bring distinct improvements in clinical efficacy. PMID- 7480068 TI - Microbiological quality of pharmaceutical raw materials. AB - A total of 115 samples of pharmaceutical raw materials (excipients) were analysed: 36 lactose, 27 talc, 19 corn starch, 18 arabic gum, 8 gelatin, 3 gelatinized starch, 3 cellulose and one tragacanth gum. 69.9% of the samples showed less than 10(2) bacteria/g (mean = 23.2 cfu/g) and 95.2% less than 10(2) fungi/g (mean = 4.92 cfu/g). Arabic and tragacanth gum were the most contaminated products by bacteria and fungi, respectively. Pregelatinized starch, cellulose and lactose were the least contaminated excipients. In none of the samples Escherichia coli or Salmonella-Shigella were detected; however, strains of Enterobacter, Serratia and Proteus were isolated from 10 samples of 5 different excipients. Only 5 samples did not comply with the microbiological standards as established by the European Pharmacopoeia and USP. PMID- 7480069 TI - Towards understanding the role of the first extracellular loop for the binding of peptide hormones to G-protein coupled receptors. AB - By a combination of molecular modelling and site-directed mutagenesis studies, we have recently identified a key residue in the first extracellular loop which determines agonist selectivity and high-affinity binding in the V1a vasopressin receptor. Based on primary sequence analysis and structure-activity relationship studies of other neuropeptides and their receptors, the corresponding amino acid in the first extracellular loop is proposed to play a homologous role in conferring affinity and selectivity. This would seem to be the case notably for angiotensin, cholecystokinin, neuropeptide Y and neurokinin receptors. PMID- 7480070 TI - Monthly prostaglandin bibliography prepared by the University of Sheffield Biomedical Information Service. PMID- 7480071 TI - Inhibition by beraprost sodium of thrombin-induced increase in endothelial macromolecular permeability. AB - Effect of beraprost sodium (BPS), a long-acting and orally active stable analogue of PGI2, on the macromolecular permeability of cultured vascular endothelial cells (HUVEC) was detected by the transport of FITC-albumin. Thrombin treatment resulted in induction of FITC-albumin transport across the endothelial cell monolayer. The albumin transport induced by thrombin was not accompanied by any damage to the cells. BPS had no effect on the permeability of resting endothelial monolayers, while BPS inhibited the thrombin-induced increase in the albumin permeability in a dose-dependent manner (30-1000 nM). Treatment of the cells with PGI2 or dibutyryl cAMP caused a significant inhibition of the thrombin-induced increase in the albumin permeability. These results strongly suggested that BPS suppressed the thrombin-induced macromolecular permeability in HUVEC through the elevation of its intracellular cAMP, and that BPS was a suppressor against inflammatory vascular changes such as exudation. PMID- 7480072 TI - The contribution of hepatocytes to prostaglandin synthesis in rat liver. AB - The contribution of hepatocytes to liver prostaglandin (PG) synthesis Is not clear. We compared prostaglandin synthesis in homogenates of whole liver, freshly isolated hepatocytes, and mixed non-parenchymal cells from the same rat livers, and optimized the assay. Whole liver homogenates made 27.2 +/- 7.1 mg PGE2/mg protein/5 min (+/- SEM, n = 4 livers). Hepatocyte homogenates made 39 +/- 9% as much PGE2/mg protein as did the matched whole livers. Non-parenchymal cell homogenates made slightly more PGE2 than whole liver, but much more PGD2. Subsequent studies showed that fresh hepatocyte suspensions contain significant contamination with non-parenchymal cells. Homogenates from ricin-purified hepatocyte monolayers made at least half as much PGE2 as did conventional monolayers. However, taking cellular purity into account, hepatocytes must contain much less than a third of liver cyclooxygenase activity. PMID- 7480073 TI - Evidence of a temperature-sensitive step in the release of prostaglandin E2 in calcium ionophore-stimulated rat muscle. AB - Recent studies have shown that mild hypothermia (32-35 degrees C) confers striking protection against ischemic muscle and neuronal injuries, although the mechanisms are unknown. We previously demonstrated that the release of prostaglandin E2 (PGE2) from metabolically stressed muscles was dependent on calcium and was abolished at or below 35 degrees C. In this study, we examined the temperature response of the release of arachidonic acid (AA) and its cyclooxygenase metabolites, PGE2 and prostaglandin F2 alpha (PGF2 alpha) from rat skeletal muscle in the presence of calcium ionophore A23187, an agent that directly elevates intracellular calcium. Calcium ionophore markedly stimulated the release of AA, PGE2 and PGF2 alpha at 37 degrees C, as expected. Reducing the temperature to 35 degrees C and below sharply decreased PGE2 and PGF2 alpha release but not AA release. The activity of phospholipase A2 stimulated by calcium ionophore was unaffected when temperature of incubation was lowered from 37 to 32 degrees C. The results suggest that reducing temperature from 37 degrees C to 35 degrees C or below inhibits the conversion from free arachidonate to PGs in calcium ionophore-stimulated muscle. PMID- 7480074 TI - The effect of a TXA2 receptor antagonist ON-579 on experimental allergic reactions. AB - The effect of a thromboxane A2 (TXA2) receptor antagonist, ON-579, on experimental allergic skin and airway reactions was studied in vivo. ON-579 at doses of 1 and 20 mg/kg clearly inhibited U-46619-induced increases in respiratory resistance (Rrs) in guinea pigs. ON-579 at doses of 1, 20 and 50 mg/kg inhibited the aerosolized antigen-induced biphasic increase in Rrs in guinea pigs. Moreover, ON-579 clearly inhibited repeated aeroantigen-induced airway hyperreactivity in guinea pigs. ON-579, however, did not have any significant effects on allergic cutaneous reactions in rats. These results suggest that ON-579 is a relatively selective TXA2 antagonist, especially in the airways, and indicate the efficacy of ON-579 on antigen-induced increase in airway resistance and antigen-induced airway hyperreactivity in guinea pigs. PMID- 7480075 TI - Protective capability of dietary Zizyphus mistol seed oil, rich in 18:3, n-3, on the development of two murine mammary gland adenocarcinomas with high or low metastatic potential. AB - The modulating effect of dietary Zizyphus mistol seed oil, rich in the n-3 polyunsaturated fatty acid alpha-linolenic acid, on two murine mammary gland adenocarcinomas having low (M2) and high (MM3) metastatic abilities was investigated. Following tumor subcutaneous inoculation, 165 BALB/c mice were fed on a semisynthetic diet containing 5% of mistol oil which has 25% of 18:3, n-3 (mistol group). The other group received a stock diet (C group). Tumor growth parameters such as success of tumor implants (ST), latency time (LT), tumor growth (TG), tumor volume (TV), survival, and number and sites of metastases were recorded. Fatty acid profiles of liver lipids correlated well with dietary fatty acids, with a n-3/n-6 ratio of 0.90:1 in the mistol group against 0.24:1 in the C group. None of the tumor growth parameters were worse in the mistol group with respect to the controls. On the contrary, survival in M2 hosts and tumor weight in MM3-bearing mice were significantly ameliorated. A tendency towards a diminution of total number of metastases in both types of tumor was observed in the mistol group. From these results, it is suggested that Z. mistol seed oil may be of potential value in nutritional oncology. PMID- 7480076 TI - Improvement in blood flow and diabetic neuropathy by thromboxane A2 dual blocker KDI-792. AB - We studied the effect of a thromboxane A2 (TXA2) dual blocker KDI-792 on skin blood flow as well as on the peripheral nerve function of nine diabetics with neuropathy. After administration of KDI-792, there was no change in urinary TXB2; however, urinary 6-keto-prostaglandin (PG) F1 alpha increased significantly. Nerve conduction velocity (NCV) and vibration perception threshold (VT) in the four extremities improved significantly, as did deep skin temperature and skin blood flow. The degree of improvement in sensory NCV in the lower extremities correlated significantly with that of deep skin temperature in the toes and the degree of improvement of VT in the lower extremities correlated well with that of deep skin temperature in the soles and of skin blood flow in the toes. Based on these findings, treatment of diabetic neuropathy with a TXA2 dual blocker appears to increase PGI2 production, improving blood flow, and resulting in improvement of nerve functions. PMID- 7480077 TI - Eicosanoids, fatty acids and neutrophils: their relevance to the pathophysiology of disease. AB - PUFA and their eicosanoid metabolites are potent biological modifiers. They have beneficial effects in a number of diseases, which may result in part from their direct actions on neutrophils as well as from their ability to modulate eicosanoid biosynthesis. A consideration of their interactions with other cell types, e.g. lymphocytes and macrophages, is beyond the scope of this review. Small alterations in structure can result in large changes in the neutrophil response. This will have important implications for the further development and use of fatty acids for therapeutic purposes. PMID- 7480078 TI - Prostaglandin synthesis in human cancer cells: influence of fatty acids and butyrate. AB - Previous research has suggested that prostaglandins (PGs) may play a role in the development of colon cancer since tumor cells produce more PGs than normal cells. However, the exact mechanism by which PGs play a role in the development of cancer is not known. In addition, factors that influence PG synthesis are not known since they are complicated by the presence of homeostatic mechanisms. To avoid the homeostatic mechanisms, the present research was designed to examine factors that may influence PG synthesis in an in vitro system, i.e., a tissue culture. We have chosen two human colon cancer cell lines that differ in their ability to metabolize long-chain fatty acids (LCFAs), LS174T cells and HT-29 cells. We examined the effect of LCFAs on their membrane fatty acid composition, growth, and ability to release the main PGs (PGE2 and PGI). The LCFAs used were those most common in the colonic lumen [18:0, 18:2 (n-6), and 18:3 (n-3)]. In addition, we examined the effect of butyrate on the above mentioned parameters. Butyrate is produced in the colon through fermentation of dietary fibers. The data obtained suggest that although both of these tumor cell lines are of human colonic origin, they differ in their response to LCFAs and butyrate in some of the characteristics studied, such as growth, composition of membranes, and the relationship between membrane FA composition and PG synthesis. Polyunsaturated fatty acid supplementation stimulated the growth of HT-29 cells but not of LS174T cells when compared with growth in media supplemented with 18:0.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7480080 TI - Monthly prostaglandin bibliography prepared by the University of Sheffield Biomedical Information Service. PMID- 7480079 TI - Effects of gamma-linolenic and dihomo-gamma-linolenic acids on 7,12 dimethylbenz(alpha)anthracene-induced mammary tumors in rats. AB - The effects of pure gamma-linolenic acid (GLA; C18:3, n-6) and dihomo-gamma linolenic acid (DGLA; C20:3, n-6) were investigated in 7,12 dimethylbenz(alpha)anthracene (DMBA) (10 mg/rat)-induced mammary tumors in Sprague-Dawley rats. 0.15 g of GLA, DGLA, or corn oil (CO) were administered (two times per week) by oral intubation, for 12 weeks to rats maintained on a 5% (w/w) CO diet. Tumor incidence, tumor multiplicity, and percent of tumor-bearing rats were highest in the CO group. Tumor multiplicity was significantly reduced in the GLA group (p = 0.015). Feeding of GLA and DGLA resulted in significant alterations in levels of these fatty acids in phospholipids of mammary tissue, thymus, colon, liver, stomach, and ovary. These results suggest that GLA may have a small, but significant, inhibitory effect on the development of DMBA-induced mammary tumors in rats. PMID- 7480081 TI - The control of prostaglandin production by the endometrium in relation to luteolysis and menstruation. AB - Oestradiol acting on a progesterone-primed uterus stimulates prostaglandin (PG) F2 alpha synthesis by the endometrium. In some species (notably the sheep, cow and goat) oxytocin released from the ovary also forms part of the physiological stimulus for increased endometrial PGF2 alpha production. The corpus luteum contains high concentrations (> 1 microgram/g tissue) of this peptide in these species. The intracellular mechanisms by which these three hormones control endometrial PGF2 alpha synthesis and release are far from clear. Oxytocin stimulates the synthesis of inositol phosphates and diacylglycerol in the endometrium of some species, but whether this pathway is involved in endometrial PGF2 alpha synthesis is still open to question. There is evidence that increased endometrial PGF2 alpha synthesis is dependent upon increased endometrial protein synthesis but, apart from the recorded effects of steroid hormones on the concentrations of phospholipase A2, prostaglandin H synthase and oxytocin receptors, it is not known what other endometrial proteins are involved. Some disorders of menstruation are associated with abnormal PG production by the endometrium, but the reasons for this abnormality are not clear. During early pregnancy an increase in PGF2 alpha synthesis by the endometrium is prevented, except in the pig where the PGF2 alpha produced is directed from the venous drainage to the uterine lumen. In those species in which endometrial PGF2 alpha synthesis is dependent upon oxytocin secreted by the ovary, the conceptus secretes an interferon-tau (previously named trophoblast protein-1) which prevents oestradiol and oxytocin acting on a progesterone-primed uterus from stimulating endometrial PGF2 alpha synthesis. The identities of the factors produced by the conceptus which prevent endometrial PGF2 alpha synthesis during early pregnancy in other species are not known, although it is clear that they are not interferons. PMID- 7480082 TI - PAF effects on eicosanoid release in neonatal rat cardiomyocytes. AB - The aim of this study was to find out whether, in neonatal rat cardiomyocytes, platelet-activating factor (PAF) can stimulate eicosanoid formation. For this purpose neonatal cardiomyocytes were incubated for 60 min at 37 degrees C in HANKS buffer with PAF (10-1000 nM), and the eicosanoids thromboxane A2 (TXA2) and prostacyclin (PGI2) were assessed in the supernatant as TXB2 and 6-keto-PGF1 alpha, respectively, by an enzyme immunoassay. PAF caused concentration-dependent release of PGI2; TXA2, however, was significantly released only at the highest concentration of PAF (1000 nM). Acetylsalicylic acid (556 microM) and the PAF antagonist WEB 2086 (10 microM) significantly attenuated PAF-induced eicosanoid formation. We conclude that in neonatal rat cardiomyocytes PAF can induce eicosanoid formation and this effect is brought about by activation of a specific PAF receptor. PMID- 7480083 TI - Distribution of arachidonic, eicosapentaenoic, docosahexaenoic and related fatty acids in ovine endometrial phospholipids in late gestation and labor. AB - The quantitative distribution of phospholipid (PL) fatty acids from ovine endometrial tissues taken at 105 (n = 3) and 131 and 147 (n = 5) days of gestation age (dGA) and in spontaneous labor (SL, n = 3) is reported. Phosphatidylcholine (PC), phosphatidylethanolamine (PE), phosphatidylinositol (PI), phosphatidylserine (PS), and lysophosphatidylcholine (LPC), were separated by thin layer chromatography (TLC) and analyzed for fatty acid composition by quantitative gas chromatography (GC). Saturates are found mainly in PS and PI and unsaturates predominantly in PC and PE. The major long-chain polyunsaturated fatty acids (LC-PUFA), arachidonic acid (AA), eicosapentaenoic acid (EPA), and docosahexaenoic acid (DHA), are found primarily in PC, PE, and PI. AA accumulates in PC, PI and PS (p < 0.05) from late gestation to term and significantly declines in PC and PS (p < 0.02) during labor, suggesting that ovine endometrium is a possible source of prostaglandin (PG) precursors. EPA decreases significantly from around 105 dGA to term and at labor in PC (p < 0.02) and in PI (p < 0.01), which may indicate the involvement of 3-series PGs in the regulation of uterine contraction. Unsaturation index (UI) and total PUFA increase from late gestation to term in PE (p < 0.05) and decrease during labor (p < 0.05). The ratios of n-6/n-3 PUFA increase in PI (p < 0.05) and in PC (p < 0.01) during labor mainly due to the decline of EPA in these PL.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7480084 TI - Consumption of a garlic clove a day could be beneficial in preventing thrombosis. AB - The effect of the consumption of a fresh clove of garlic on platelet thromboxane production was examined. A group of male volunteers in the age range 40-50 years participated in the study. Each volunteer consumed one clove (approximately 3 g) of fresh garlic daily for a period of 16 weeks. Each participant served as his own control. Thromboxane B2 (TXB2, a stable metabolite of thromboxane A2), cholesterol and glucose were determined in serum obtained after blood clotting. After 26 weeks of garlic consumption, there was an approximately 20% reduction of serum cholesterol and about 80% reduction in serum thromboxane. No change in the level of serum glucose was observed. Thus, it appears that small amounts of fresh garlic consumed over a long period of time may be beneficial in the prevention of thrombosis. PMID- 7480085 TI - Effect of BAY u3405, a thromboxane A2 receptor antagonist, on neuro-effector transmission in canine tracheal tissue. AB - Thromboxane A2 (TXA2) is reported to potentiate vagal nerve neuro-effector transmission in airway smooth muscle tissue. We investigated the effects of BAY u3405 (3(R)-[[4-fluorophenyl)sulfonyl]amino]-1,2,3,4,-tetrahydro-9H-carbazole - 9 propanoic acid), a potent and selective TXA2 receptor antagonist, on the increase in vagal nerve neuro-effector transmission induced by a TXA2 mimetic, U-46619, in the canine trachea. We measured the contractions of canine tracheal smooth muscle evoked by electrical field stimulation (EFS) and by acetylcholine (ACh) in the presence and absence of a subthreshold dose of U-46619 (the highest dose that did not induce any smooth muscle contraction). We then examined whether BAY u3405 inhibited the effect of U-46619 on tracheal smooth muscle. The following results were obtained: (i) subthreshold doses of U-46619 (10(-10) M and 10(-9) M) significantly increased the amplitude of the contractions evoked by EFS; (ii) by contrast, U-46619 had no effect on the contractile response of smooth muscle to exogenously applied ACh; (iii) the contraction evoked by EFS was completely abolished by the application of atropine (10(-6) M) or tetrodotoxin (10(-7) M), indicating that EFS caused the smooth muscle contraction through the release of ACh from vagal nerve terminals; and (iv) pretreatment with BAY u3405 (10(-6) M) abolished the excitatory action of U-46619 on the amplitude of twitch contraction evoked by EFS in the trachea. These results indicate that U-46619, at low concentrations, has a prejunctional action stimulating neuro-effector transmission, presumably increasing ACh release from vagal nerve terminals through TXA2 receptors.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7480086 TI - Acute burn down regulates rabbit splanchnic and renal prostanoid release. AB - This study examines the hypothesis that acute thermal injury decreases renal and splanchnic vasodilator eicosanoids. Anesthetized rabbits were subjected to sham or a 25% total body surface area burn and fluid resuscitated. At 2, 4, 6, 12, and 24 h postburn the superior mesenteric and renal arteries were cannulated and perfused in vitro with their end organs with Krebs buffer (pH 7.4, 37 degrees C). Renal and splanchnic prostaglandins (PGs) 6-keto-PGF1 alpha (PGI2), and PGE2, and thromboxane B2 (TxB2) release were measured by EIA at 15 min of perfusion. The major eicosanoids released were PGI2 from the splanchnic bed and PGI2 and PGE2 from the kidney. Renal PGE2 and PGI2 and splanchnic PGI2 release were decreased by 50% or more 12 h postburn (p < 0.01) but were restored to sham burn levels 24 h postburn. Loss of these endogenous renal and splanchnic vasodilators 12 h postburn may contribute to ischemia of both vascular beds at this critical time period following acute burn injury. PMID- 7480087 TI - EGF and TPA stimulate de novo synthesis of PGHS-1 and PGHS-2 through different signal transduction pathways. AB - Epidermal growth factor (EGF) as well as phorbol 12-myristate 13-acetate (TPA) stimulate de novo synthesis of PGHS (prostaglandin H synthase)-1 and PGHS-2 mRNA, resulting in increased production of PGE2 in rat tracheal epithelial cells (RTE, EGV-6 cells). Stimulation of PGE2 production by TPA is more potent than that by EGF. Staurosporine and H-7, protein kinase C (PKC) inhibitors, suppressed the increase of mRNA and PGE2 levels caused by TPA, but not that caused by EGF. On the other hand, methyl 2,5-dihydroxycinnamate, a tyrosine kinase inhibitor (TKI), suppressed the increase of mRNA and PGE2 levels caused by EGF, but not that caused by TPA. These results indicate that EGF stimulates de novo synthesis of PGHS-1 and PGHS-2 mRNA through a signal transduction pathway which is independent from PKC-associated mechanisms but dependent upon the tyrosine kinase activity of the EGF receptor. PMID- 7480089 TI - [The leukotriene signal pathway--a very promising point of entry for inhibitors in therapy of inflammatory diseases]. PMID- 7480088 TI - [Receptor independent activation of G proteins]. PMID- 7480091 TI - [1994 annual report on the development of the HIV-AIDS epidemic in Germany]. PMID- 7480090 TI - [Pharmacology and society]. PMID- 7480092 TI - Marine fungi--a prolific resource of biologically active natural products? PMID- 7480093 TI - Application of the rough sets theory to the analysis of structure-activity relationships of antimicrobial pyridinium compounds. AB - Relationship between chemical structure and antimicrobial activity of 80 quaternary pyridinium chlorides were analysed using the method of rough sets. The compounds are described by eight conditions attributes (substituents in pyridine) and divided into three classes of activity (MIC). Using the rough sets approach a smallest set of four condition attributes significant for a high quality of classification and accuracy of classes has been found. The resulting decision rules describe relations between structure and antibacterial activity of pyridinium chlorides in terms of significant condition attributes. This may be helpful in predicting structures of new antibacterial compounds to be synthesized. PMID- 7480094 TI - Determination of itraconazole and its metabolite in plasma using SPE-HPLC. AB - A SPE-HPLC procedure for the determination of itraconazole and its metabolite has been developed. The method was shown to be suitable for routine therapeutic monitoring in leukemic patients. PMID- 7480095 TI - A new sensitive and simple method for detection of catecholamines from adrenal chromaffin cells. AB - A biosensor was used for the analysis of catecholamines in media and lysates of cultured bovine adrenal chromaffin cells. The sensor is composed of coimmobilised laccase and glucose dehydrogenase coupled with an oxygen electrode, using the catalytic effect of cate cholamines for glucose oxidation in this system. The analysis time is almost 5 min. The correlation between the biosensor and HPLC determination is 0.99. PMID- 7480096 TI - Influence of ointment formulation on skin erythema induced by nicotinate esters. AB - The influence of the chemico-physical nature of the vehicle on the skin penetration of topically applied drugs was evaluated in vivo. Five different ointment formulations containing nicotinate esters as model penetrants were tested on human skin. The degree of drug penetration allowed by the various formulations was revealed by means of the erythema induced by the drug, detected by a X-Rite tristimulus reflection colorimeter. The excipient influenced the penetration of the nicotinate esters used to various extents. The concentrations of the tested drugs were found to be an important factor influencing drug penetration and persistence of erythema. PMID- 7480097 TI - Pharmacokinetic interaction between endogenous cortisol and exogenous corticosteroids. AB - A problem in the evaluation of pharmacokinetic interactions between prednisolone and cortisol is that both steroids bind to cortisol binding globulin (CBG) and albumin. The binding of both steroids to CBG is saturable in the therapeutic concentration range. General drug binding equations were applied to two drugs and two binding sites and two implicit cubic equations were derived. These equations cannot be solved algebraically. However, the free concentration can be calculated using spreadsheet programs on a personal computer. A spreadsheet for these equations was developed using EXCEL 5.0 and its SOLVER option. Free concentrations of prednisolone and cortisol were determined as a function of total concentrations of both ligands. The simulations show that the protein binding of cortisol remains relatively constant in the physiological range but changes when exogenous corticosteroid is present. However, the degree of protein binding of exogenous corticosteroids that bind to CBG depends on the cortisol concentration which competes for binding sites. At the same time, the exogenous corticosteroid suppresses the release of endogenous cortisol. The presented approach is able to take all of these pharmacokinetic and pharmacodynamic interactions into account leading to a more accurate estimation of active free corticosteroid concentrations. PMID- 7480098 TI - Pharmacokinetic interactions of nifedipine and quinidine. AB - Several clinical investigations have been published regarding the interaction of nifedipine and quinidine. The results of these studies are contradictory. In vitro studies indicate that the 3-hydroxylation and N-oxigenation of quinidine appear to involve the P4503A4 family, a form of cytochrome that predominantly catalyzes the aromatization of nifedipine, too. The aim of our study was to investigate the effect of oral intake of 200 mg quinidine on the kinetics of 20 mg nifedipine as a retarded formulation and vice versa. Twelve healthy male volunteers between 18 and 40 years were treated. Each subject was studied on three occasions each separated by a one week washout period. Drug administration consisted of one oral dose of nifedipine (Adalat retard 20 mg), one oral dose of quinidine (Chinidin sulfuricum "Buchler" 200 mg) or a combination of both (20 mg nifedipine and 200 mg quinidine) in a randomised 3 way crossover. Administration of the test drugs in combination slightly increased the bioavailability of both- nifedipine [N] to 18% and quinidine [Q] to 16%--and decreased the clearance of both drugs. The results were not statistically significant. Based on our data, the combination of nifedipine and quinidine seems to lack a clinically relevant interaction. PMID- 7480099 TI - Effect of doxorubicin and phenytoin on sea urchin development. AB - The effect of doxorubicin and phenytoin on Paracentrotus lividus development and fertilization is reported. Doxorubicin at concentrations between 1.4 and 1.8 microM causes severe disturbances of development and at 2.0 microM, complete embryonic degeneration, if added in the period from fertilization on. Concentrations up to 20 microM are ineffective on development if the treatment is started at 12 h after fertilization. Phenytoin at concentrations between 4.0 and 7.0 microM causes severe disturbances of development, if added from 5 min after fertilization on. Concentrations of 10 microM are lethal to all embryos. Again, if the drug is added at 12 h after fertilization, no effect is observed up to the concentration of 20 microM. Both drugs are ineffective on the fertilization process up to the concentration of 20 microM for doxorubicin and of 200 microM for phenytoin. PMID- 7480100 TI - Investigation of cytotoxic effects of 8 norbornane derivatives on 4 human cancer cell lines using the MTT assay. AB - A group of 8 structurally modified norbornane analogs, selected on basis of water solubility, was tested for its potential cytotoxic effects using a modified MTT assay. Four tumor cell lines, three hematological and a hepatocellular one were used as an experimental model system. Some analogs exhibited an antitumor effect which was shown to be tissue specific. The cytotoxic effect is most likely due to the nature of the side chains rather than to the basic bicyclic norbornane structure of the molecule. These results not only provide a basis for the synthesis of derivatives with effective antitumor activity, but also give some indication of the the molecular target of these compounds. PMID- 7480101 TI - Local anesthetic effect of carbisocaine and its enantiomers. AB - The optically active isomers of carbisocaine [1-methyl-2-diethylaminoethyl ester of 2-(n)-heptyloxycarbonilic acid] were prepared. The blocking activity of equimolar concentrations of the carbisocaine and its corresponding enantiomers was tested on isolated rat sciatic nerves. There were no significant differences between the anesthetic action of racemic form and enantiomers, however, lower activity for the (--)-enantiomer was observed. The results may indicate negligible stereoselectivity of action of highly lipophilic local anesthetic carbisocaine in the excitable membrane. PMID- 7480103 TI - Arylhydrazones and phenylsemicarbazones of 2-aminobenzophenones as antithrombotic agents. PMID- 7480102 TI - [Comparison of the bioavailability of beta-aescin after single oral administration of two different drug formulations containing an extract of horse chestnut seeds]. AB - The relative oral bioavailability of beta-escine (CAS 11072-93-8) from a sugar coated tablet formulation was compared to a reference preparation available in capsule form in 18 healthy, male volunteers over a 48 h period. The study design was randomized, single-blind and cross-over. Both the test and the reference preparation contained 50 mg standardized horse chestnut seed extract; beta-escine was taken as the reference substance. By means of a newly developed, validated radioimmunosorbent assay (RIA), beta-escine in plasma was determined (blind samples) after oral intake of a single dose of each drug formulation. The confidence limits calculated for the AUC, Cmax and Tmax of the test preparation exceed the upper limit of the specified equivalence range of 80%--125%, but do never fall below the lower limit. Therefore, bioin-equivalence cannot be rejected statistically. All the bioavailability data for the test preparation--measured with the newly developed RIA--exceed the corresponding values for the reference preparation. As the rate of absorption of aesculetinic triterpene glycosides is low, the higher bioavailability of the test preparation is desirable from a therapeutical point of view. Since the reference preparation is classified as being clinically effective, the test preparation must also be estimated as being clinically effective. Adverse drug effects were not observed with either the test preparation or the reference preparation. PMID- 7480104 TI - Biological accessibility and distribution of calcitonin in rats with experimental osteoporosis. PMID- 7480105 TI - [The metabolism of the beta receptor blocker talinolol in humans--relationship between structure, polarity and amount excreted]. PMID- 7480107 TI - The Anna-Monika-Foundation--an endeavor in depression research. PMID- 7480106 TI - Fast negative feedback actions of corticosterone and of dexamethasone on CRF stimulated ACTH secretion in rats. PMID- 7480108 TI - Norepinephrine metabolism and psychoactive drugs in the endogenous depressions. 1968. AB - After intraventricular injection of norepinephrine-H3, the concentration of norepinephrine, of normetanephrine and of the deaminated catechols in rat brains was determined, following action of imipramine, desmethylimipramine, chlorpromazine, lithium chloride or cocaine. Following administration of imipramine, desmethylimipramine, and chlorpromazine, norepinephrine concentration decreased significantly at first, had distinctly increased 4.5 hours after imipramine and desmethylimipramine but was normal once again after chlorpromazine. Normetanephrine concentration increased after imipramine and desmethylimipramine but was unchanged after chlorpromazine. Under the effect of these drugs, the deaminated catechols showed no changes compared with control values. Cocaine resembled the antidepressants, but the amount of deaminated compounds was reduced. Lithium chloride, on the other hand, increased the concentration of deaminated catechols under certain conditions, reduced normetanephrine concentration but did not influence norepinephrine concentration. In addition to the animal experiments, the following data of six patients with an "endogenous" depression were recorded over a period of several weeks: the clinical findings by means of the Hamilton Depression Rating Scale, and the excretion of normetanephrine and of vanillylmandelic acid (VMA) in the urine before, during and after treatment with imipramine. The therapy led to a significant reduction of VMA; however, this reduction cannot be correlated with an improvement in the clinical findings. On the other hand, excretion of normetanephrine is apparently not dependent on the administration of imipramine but seems to reflect the clinical state, since improvement of the depression was regularly combined with an increased excretion of normetanephrine. PMID- 7480109 TI - The clinical research of affective disorders. A personal perspective. PMID- 7480110 TI - Evolutionary radiation of visual and olfactory brain systems in primates, bats and insectivores. AB - How brains have evolved in response to particular selection pressures is illuminated by ecological correlates of differences in brain structure among contemporary species. The focus of most comparative studies has been on the overall size of brains relative to body size, hence ignoring the ways in which selection operates on specific neural systems. Here we investigate evolutionary radiations in the size of visual and olfactory brain structures within three orders of mammals: primates, bats and insectivores. The comparative relationships within these three orders show both similarities and differences. After removal of the allometric effect of overall brain size, the sizes of different structures within each sensory modality are positively correlated in all three orders. Correlations between visual and olfactory structures, however, are negative in primates, negative but non-significant in insectivores, and positive in bats. In both primates and insectivores, nocturnal lineages tend to have larger olfactory structures than do diurnal or partly diurnal lineages, and among the primates diurnal lineages have larger striate visual cortexes. Hence the apparent trade off between vision and olfaction in primates seems to be related to the divergence of nocturnal and diurnal forms. However, negative correlations between visual and olfactory structures were also found when nocturnal strepsirhines and diurnal haplorhines were analysed separately, suggesting that ecological variables in addition to activity timing may be significant. Indeed, there were also associations with diet: frugivory was associated with enlargements of the geniculostriate visual system in diurnal primates, enlargements of olfactory structures in nocturnal primates, and possibly enlargements of both in bats. Further ecological associations were found within insectivores: aquatic lineages had smaller olfactory structures than in their non-aquatic counterparts, and fossorial lineages had smaller optic nerves than in non-fossorial forms. We conclude that activity timing, diet and habitat have each played a role in the evolutionary radiation of mammalian sensory systems, but with varying effects in the different taxa. Some of the associations between ecology and sensory systems suggest alternative explanations for correlates of overall brain size, which have in the past commonly been interpreted in terms of selection on intelligence. PMID- 7480111 TI - The spatial struggle of tit-for-tat and defect. AB - The pioneering work by Trivers (1971), Axelrod (1984) and Axelrod & Hamilton (1981) has stimulated continuing interest in explaining the evolution of cooperation by game theory, in particular, the iterated prisoner's dilemma and the strategy of tit-for-tat. However these models suffer from a lack of biological reality, most seriously because it is assumed that players meet opponents at random from the population and, unless the population is very small, this excludes the repeated encounters necessary for tit-for-tat to prosper. To meet some of the objections, we consider a model with two types of players, defectors (D) and tit-for-tat players (T), in a spatially homogeneous environment with player densities varying continuously in space and time. Players only encounter neighbours but move at random in space. The analysis demonstrates major new conclusions, the three most important being as follows. First, stable coexistence with constant densities of both players is possible. Second, stable coexistence in a pattern (a spatially inhomogeneous stationary state) may be possible when it is impossible for constant distributions (even unstable ones) to exist. Third, invasion by a very small number of T-players is sometimes possible (in contrast with the usual predictions) and so a mutation to tit-for-tat may lead to a population of defectors being displaced by the T-players. PMID- 7480112 TI - A composite estimate of primate phylogeny. AB - This paper presents an estimate of the phylogeny of all 203 species of primate. The composite tree is derived by applying a parsimony algorithm to over a hundred previous estimates, and is well resolved, containing 160 nodes. The ages of over half the clades in the tree have been estimated from information in the literature. Bootstrapping has been used to indicate the degree of certainty associated with each clade. The tree will be a useful framework for comparative biologists and shows which areas of primate phylogeny are still only sketchily known. PMID- 7480113 TI - Abundant distribution of locustatachykinin-like peptide in the nervous system and intestine of the cockroach Leucophaea maderae. AB - An antiserum raised to the locust neuropeptide locustatachykinin I (LomTK I) was used for analysis of the distribution of tachykinin-related peptide in the cockroach Leucophaea maderae. Extracts of dissected brains, suboesophageal ganglia, thoracic ganglia and midguts were separated by high performance liquid chromatography and the fractions analysed in enzyme-linked immunosorbent assay with use of the LomTK antiserum. Each of the tissues was found to contain LomTK like immunoreactive (LomTK-LI) components with retention times corresponding approximately to synthetic LomTK I and II and callitachykinins I and II. The LomTK antiserum was also used for immunocytochemical mapping of peptide in the nervous system and intestine of L. maderae. A large number of LomTK-LI interneurons were detected in the proto-, deuto- and tritocerebrum of the brain and in the suboesophaegeal ganglion. The immunoreactive neurons supply processes to most parts of the brain: the central body, protocerebral bridge, mushroom body calyces, antennal lobes, optic lobe and most regions of the non-glomerular neuropil. A few protocerebral neurons send LomTK-LI processes to the glandular lobe of the corpora cardiaca. In each of the thoracic ganglia there are six LomTK LI interneurons and in each of the unfused abdominal ones there are two interneurons. The fused terminal ganglion contains some additional cell bodies in the posterior neuromers. LomTK-LI cell bodies were detected in the frontal ganglion and fibres were seen in this ganglion as well as in the hypocerebral ganglion. The frontal ganglion supplies LomTK-LI processes to the muscle layer of the pharynx. The muscle layer of the midgut is innervated by LomTK-LI fibres from the stomatogastric system (oesophageal nerve and associated ganglia). Additionally the midgut contains numerous LomTK-LI endocrine cells. A number of the pharyngeal dilator muscles were also found to be innervated by LomTK-LI fibres, probably derived from cell bodies in the suboesophageal ganglion. All the LomTK-LI neurons of the central nervous system appear to be interneurons, suggesting a neuromodulatory role of the endogenous tachykinins. The tachykinin like peptides from peripheral ganglia may be involved in the control of foregut and midgut contractility and possibly the peptide of the endocrine cells in the midgut has additional actions related to intestinal function. PMID- 7480114 TI - Integrating sphere effect in whole-bladder-wall photodynamic therapy: II. The influence of urine at 458, 488, 514 and 630 nm optical irradiation. AB - Whole-bladder-wall (WBW) photodynamic therapy (PDT) performed with 458 nm instead of 630 nm wavelength might be advantageous. On the basis of Monte Carlo (MC) computer simulations using in vitro bladder optical properties, these wavelengths show an equally strong integrating sphere effect, while haematoporphyrin derivatives can be excited equally efficiently and more easily with an Ar+ laser at 458 nm. To test this, fluence rates were measured at the walls of two piglet bladders during in vivo and in vitro WBW optical irradiations at 458, 488, 514 and 630 nm. In the in vitro experiment, a controlled amount of urine with known absorption coefficient at the irradiation wavelengths was introduced in the bladder cavity. The optical absorption and scattering coefficients and anisotropy factor of the tissue of both piglet bladders were determined in vitro with a double integrating sphere set-up. MC simulations, using the in vitro optical properties, agree only partly with the measured bladder wall fluence rates. In the in vitro experiment with saline in the bladder cavity, the fluence rate at the bladder wall is lowest for 514 nm irradiation and highest for 458 and 630 nm irradiation. In the in vivo experiment and the in vitro experiment with light absorbing urine in the bladder cavity, which mimics the clinical situation, irradiation at 458 nm wavelength resulted in the lowest fluence rate for a given optical power emitted. It cannot be completely ruled out that in an in vivo bladder the light absorption by haemoglobin further reduces the integrating sphere effect at wavelengths shorter than 630 nm. Thus, WBW PDT with red light (630 nm) is technically more advantageous than that with green light (514 nm) or blue light (488 and 458 nm) as this gives the strongest integrating sphere effect. PMID- 7480115 TI - Dosimetry of 169 Yb seed model X1267. AB - Unlike previous brachytherapy sources a number of published studies have been addressed to the dosimetry of 169 Yb seeds, manufactured in several prototypes, before widespread clinical use has been made. Discrepancies seen in the dosimetry obtained for ytterbium seed prototypes appear to be related to inconsistency and non-reproducibility in the vendor's calibration procedure to determine contained activity. Av. The comparison of 169 Yb seed dosimetries demonstrates a need for more accurate implementation of calibration procedures to determine the air kerma rate for the definitive 169 Yb seed design. This paper reports an experimental procedure to determine the reference air kerma rate, Kr (mu Gy h-1), defined as the kerma rate at 1 m along the source transverse axis in free space for the new 169 Yb seed, model X1267. A mean value of the ratio Kr/Av = 1.53 mu Gy h-1 mCi-1 was obtained from determining the Kr value of eleven seeds. Since this ratio is only 3% less than the air kerma rate constant for the 169 Yb point source, (gamma delta)k = 1.58 mu Gy h-1 m2 mCi-1, this means that the Av is closer to an apparent activity than a contained activity, Ac. A Monte Carlo simulation to determine the ratio between reference air kerma rate and the contained activity gave Kr/Ac = 1.33 mu Gy h-1 mCi-1. For the dose rate constant in water we obtained DKr (1, pi/2) = 1.20 +/- 0.05 cGy h-1 (mu Gy h-1)-1, using calibrated thermoluminescent dosimeters (TLDs) and DKr (1, pi/2) = 1.21 +/- 0.03 cGy h-1 (mu Gy h-1)-1 by Monte Carlo simulation. TLDs were used both to determine the radial dose distribution along the seed transverse axis and to calibrate GAFChromic films to obtain the two-dimensional dose distribution around the seed. PMID- 7480116 TI - Implantation guidelines for 169 Yb seed interstitial treatments. AB - The adequacy of an interstitial implant carried out with a new radioactive source, the 169 Yb seed model X1267, has been examined by computing volumetric indices based on dose-volume histograms. The comparison of these indices with the ones computed for 125I seed implantations shows that the use of ytterbium seeds presents an improvement of the dose homogeneity in interstitial implants. This is due to the significant build-up associated with 169 Yb photons that reduces the rapid dose fall-off with the distance from the source. Moreover, relative to 192Ir, the lower photon energy gives 169 Yb the advantage in clinical use of reduced radiation exposure (i) to health care workers, (ii) to relatives of treated patients and (iii) to healthy neighbouring tissues of the patients if appropriate thin shielding is used. PMID- 7480117 TI - Deduction of the air w value in a therapeutic proton beam. AB - Utilization of air-filled ionization chambers with 60Co-based reference calibrations in proton dosimetry requires application of water to air stopping power ratios and the mean energy required to produce an ion pair (W or w). Accepted uncertainties in current w values for protons leads to a dosimetric uncertainty of 4 per cent when ionization chambers are employed to measure absorbed dose. For this reason, proton dosimetry protocols recommend the use of calorimetry as the absorbed dose standard. We used calorimetry in conjunction with an ionization chamber with 60Co reference calibrations to deduce the proton w value in the entrance region of a 250 MeV proton beam: 34.2 +/- 0.5 eV. Application of this w value, with its 1.5 per cent uncertainty, allows determination of dose in therapeutic proton beams, with uncertainties comparable to photon and electron values. PMID- 7480119 TI - Light propagation through teeth containing simulated caries lesions. AB - The methods currently utilized in dentistry to detect caries lesions have their limitations and alternatives are being investigated. A promising option is tooth transillumination which is based on an increase of light scattering or light absorption in the affected tissue region. In this study transillumination applied to detect approximal caries lesions was investigated using premolar teeth containing simulated caries lesions. Cavities were drilled at the approximal surface and filled with light absorbing and light scattering fluids in different dye and particle concentrations to model successive stages of lesion progress. For light absorbing cavities the extinction as function of the decadic absorption coefficient measured at the occlusal surface could be approximated by the Lambert Beer law (r = 0.98 +/- 0.01). For light scattering cavities the extinction as a function of the decadic reduced scatter coefficient wad fitted to a straight line (r = 0.98 +/- 0.03) for mu's (lambda = 633 nm) < 1.25 mm-1. For higher reduced decadic scatter coefficients the curves levelled off due to multiple scattering. In addition, the contribution of the dentinal cavity part to the radiance change induced by the total cavity was estimated. For light absorbing cavities illuminated with red light the average contribution was 10.5 (SD 4.2)% and for those illuminated with green light it was 1.4 (SD 0.9)%, indicating that the radiance change caused by a caries lesion is mainly determined by the enamel lesion part. PMID- 7480118 TI - A new correction method for gamma camera non-uniformity due to energy response variability. AB - We present a new uniformity correction (Fourier energy correction) which is designed to correct for gamma camera non-uniformity caused by variations of the energy response function within a wide spectral range. A convolution model is used to describe the spatial distortions of the energy response function. The model is solved in Fourier space. A preliminary flood acquisition is required to obtain energy-dependent Fourier weights which are used to correct subsequent acquisitions. The influence of the parameters involved in the correction procedure is studied and the Fourier energy correction is compared to a conventional multiplicative energy correction for different acquisition geometries. The Fourier energy correction appears especially useful when the energy information associated with each detected photon is analysed using a fine sampling, or when windows different from the photopeak window are used. PMID- 7480120 TI - Depth of ionization chamber in water. AB - The device developed by the authors and described here enables the user to measure the depth from the water surface to the point of measurement for a cylindrical ion chamber with a waterproof plastic cap in a water phantom, free of surface-tension error with a high precision. The device seeks vertical orientation and provides the convenience of hands-free operation. The measurement process is simple and quick with a precision of 0.1 mm. (The device is currently available as a 'water phantom depth gauge' from Nuclear Associates, Division of Victoreen Inc., Clare Place, NY, USA.) PMID- 7480121 TI - A comparison of the threshold detail detectability of a screen-film combination and computed radiology under conditions relevant to high-kVp chest radiography. AB - There is conflicting evidence regarding the viability of photostimulable phosphor computed radiology (CR) when used with high x-ray beam energies. In this study a standard screen-film combination was compared with CR using identical exposure conditions, which simulated those of a high-energy chest examination. The comparison was made using a psychophysical technique in which the threshold contrast detail detectability is assessed by human observers. Threshold contrast determination is an efficient way with which to assess the overall image quality. The CR images were processed and displayed using two independent image processing algorithms and hard-copy formats. The results showed, at the 5% level of statistical significance, that there was no difference in the overall image quality between any of the modalities being compared. It may be concluded therefore that CR is a potentially viable option for use in high-energy radiography of the chest. PMID- 7480122 TI - Inspiratory muscle training in the patient with neuromuscular disease. AB - Pulmonary complications due to respiratory muscle dysfunction are commonly a source of morbidity and mortality in patients with neuromuscular diseases. This review discusses the adverse effects of respiratory muscle weakness on pulmonary mechanics and examines the role that inspiratory muscle training may play in reversing pulmonary dysfunction in these individuals. In asymptomatic persons, it is well established that the inspiratory muscles can be trained to increase both force and endurance. In patients with neuromuscular diseases, the effects of training protocols on force and endurance are more controversial. This article reviews seven studies that have evaluated respiratory muscle training in a total of 75 patients with varied neuromuscular disorders. Training regimens included breathing through inspiratory resistive loads and isocapnic hyperpnea. Despite methodologic differences among studies, investigators have generally shown that the inspiratory muscles are similar to other skeletal muscle groups in that they can be trained for both force and endurance in these patients. The training related improvements in inspiratory muscle performance are more pronounced in patients who are less severely affected by their disease. In those patients who have disease to the extent that they are already retaining carbon dioxide, there is little change in force or endurance with training. In these individuals, the inspiratory muscles may already be working at a level sufficiently severe to provide a training stimulus with each breath. No adverse effects of inspiratory muscle training were reported. Inspiratory muscle training can improve force and endurance in patients with neuromuscular weakness.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7480123 TI - Specialization, certification, turf, and society. PMID- 7480124 TI - Validity of the Peabody Developmental Gross Motor Scale as an evaluative measure of infants receiving physical therapy. AB - BACKGROUND AND PURPOSE: The purpose of this study was to examine the validity of the Peabody Developmental Gross Motor Scale (PDMS-GM) as an evaluative measure of infants receiving physical therapy. SUBJECTS AND METHODS: Infants who attended an early intervention program (N = 124) were grouped by diagnosis: cerebral palsy, Down syndrome, hydrocephalus, preterm with developmental delay, full term with developmental delay, and other. The PDMS-GM was administered to each infant three times over a 6-month period by a therapist who did not provide treatment. RESULTS: Mean scaled scores and age-equivalent scores increased for each group. Individual change was examined using the reliable change index. The results indicated that the change in total raw score for 62% of the infants was greater than what could be attributed to measurement error. When minimal clinically important change was defined as 10 scaled score points, the index of responsiveness was equal to 0.5. This finding indicates that a sample size of 68 subjects per group would be needed when the PDMS-GM is used as an outcome measure in research. CONCLUSION AND DISCUSSION: The mean change scores for each group support the use of the PDMS-GM as an evaluative measure. For many infants, particularly infants with cerebral palsy, the PDMS-GM was not responsive to change over a 6-month period. The index of responsiveness suggests that the PDMS GM should be used only as an outcome measure in large clinical trials. The PDMS GM is not recommended for evaluating the direct effects of physical therapy but is recommended for providing a global measure of change in motor development as part of a multidimensional assessment. PMID- 7480125 TI - The effect of solid ankle-foot orthoses on movement patterns used in a supine-to stand rising task. AB - BACKGROUND AND PURPOSE: Within dynamical pattern theory, ankle motion can be proposed to be a control variable, and solid ankle-foot orthoses (SAFOs) can be considered a constraint to ankle movement. The purpose of this study was to examine the effect of SAFOs on movement patterns used to rise from the supine position to erect stance. SUBJECTS: Thirty-nine nondisabled young adults, ranging in age from 20 to 28 years (X = 22.7, SD = 1.87), participated. METHODS: Subjects were videotaped while rising from a supine position on a floor mat. Each subject performed 10 trials under each of four conditions: without SAFOs, right SAFO, left SAFO, and bilateral SAFOs. Movement patterns were described within three body components (ie, upper extremities, axial region, and lower extremities) by determining the mode and the incidence of each movement pattern under each condition. The subjects' mode movement patterns in the no SAFO condition were compared with mode movement patterns in the SAFO conditions using McNemar tests. RESULTS: Without SAFOs, subjects rose most commonly using a push and reach pattern of the upper extremities, a forward with rotation pattern in the trunk, and an asymmetrical squat in the lower extremities. Changes in the incidence of movement patterns occurred in all of the SAFO conditions when compared with the no SAFO condition. These changes resulted in more asymmetry when SAFOs were worn, and asymmetry was most notable in the axial region. CONCLUSIONS AND DISCUSSION: From a dynamic pattern theory perspective, ankle motion is a control variable for the supine-to-stand rising task. PMID- 7480126 TI - Outcomes research: shifting the dominant research paradigm in physical therapy. AB - This article discusses outcomes research in physical therapy and places its conceptual roots within the work on quality-of-care assessment. An argument is advanced that the outcomes research movement in medicine has stimulated clinical researchers in physical therapy to address disability outcomes in addition to traditional impairment outcomes. If physical therapy clinical research moves beyond this broadening of clinical outcomes to investigate explicitly the hypothesized relationship between impairment and disability, outcomes research will have stimulated a shift in the dominant research paradigm in the profession. The development and testing of theory regarding the pathogenesis of disability will be needed to guide the direction of this type of physical therapy research. Such a shift in the dominant research paradigm in physical therapy could produce dramatic findings that have a direct impact on clinical practice. PMID- 7480128 TI - Clinical assessment of the respiratory muscles. AB - This review examines approaches to evaluation of the respiratory muscles and describes new techniques that may be more quantitative, less effort dependent, and less invasive than conventional methods. To evaluate strength of the respiratory muscles, maximum inspiratory and expiratory pressures remain useful measures. Potential methodologic errors, however, necessitate careful technique. Evaluation of the twitch response to direct phrenic nerve stimulation may ultimately prove more quantitative and less effort dependent than measurements of maximum pressure. Many techniques are also available to measure endurance of respiratory muscles, but most are less than satisfactory outside the research environment because of poor reproducibility and other procedural difficulties. The maximum incremental resistive loading test, however, has proven to be practical and well tolerated. There is little substitute for careful clinical observation of respiratory muscle coordination and movement, particularly in the patient with suspected respiratory muscle weakness or chest wall distortion. In conclusion, though the respiratory muscles are difficult to evaluate, techniques are available that can be quite helpful for assessment, particularly in response to interventions such as rehabilitation. PMID- 7480127 TI - Considerations when testing and training the respiratory muscles. AB - The purpose of this review is to describe the muscles of respiration, mechanisms that lead to their dysfunction, and unique features of the respiratory muscles that should be considered when designing testing and training protocols for patients with respiratory compromise. The diaphragm's unique shape is an important determinant of its function. Respiratory pathophysiology combined with systemic abnormalities may result in inspiratory muscle weakness, fatigue, and injury. The specificity of training principle, as it applies to respiratory muscle training in patients with respiratory compromise, will be described. Precautions related to this type of training will also be outlined. In conclusion, a better understanding of the mechanisms contributing to inspiratory muscle dysfunction will assist physical therapists in designing the most appropriate training programs. This article, along with the other articles in this focus on respiratory muscle training, will provide therapists with scientifically based guidelines for rehabilitation of patients with impaired respiratory muscle function. PMID- 7480129 TI - Respiratory muscle training for patients with chronic obstructive pulmonary disease. AB - The purpose of this article is to describe respiratory muscle training techniques and the effectiveness of this training in patients with chronic obstructive pulmonary disease (COPD). The respiratory muscles can be strengthened, but the benefit of stronger respiratory muscles in patients with COPD is not clear. Maximal sustained voluntary ventilation, inspiratory resistive breathing, and threshold loading are the three most commonly used techniques for improving the endurance of the inspiratory muscles in patients with COPD. Recent studies using inspiratory resistive breathing with targeted devices or threshold trainers have shown more consistent increases in inspiratory muscle function and exercise tolerance than studies using other techniques. Endurance exercise involving the extremities improves inspiratory muscle endurance in younger individuals with cystic fibrosis but not in older persons with COPD. Recommendations are outlined regarding the techniques to use for respiratory muscle training and which patient populations may benefit from these techniques. PMID- 7480130 TI - The role of urocanic acid in UV-induced immunosuppression: recent advances (1992 1994). AB - Cis-urocanic acid (UCA), formed in the epidermis by UV irradiation of trans-UCA, has been implicated as a mediator of the immunosuppression induced by UV exposure of the skin. This review covers recent work in which the wavelength dependence of cis-UCA formation, the interaction of UCA isomers with DNA, the effects of UCA isomers on the immune system and their interaction with histamine are examined. Results are frequently conflicting, particularly when considering the possible mode of action of cis-UCA but, overall, a multifaceted role for UCA in immunomodulation by UV radiation is substantiated. PMID- 7480131 TI - p53 gene mutations and photocarcinogenesis. PMID- 7480132 TI - Formation of 7,8-dihydro-8-oxoguanine in the 1,2-dioxetane-induced oxidation of calf thymus DNA: evidence for photosensitized DNA damage by thermally generated triplet ketones in the dark. AB - Isolated calf thymus DNA was treated with the 1,2-dioxetanes 3-acetoxymethyl 3,4,4-tri-methyl-1,2-dioxetane, 2,3-dimethylbenzofuran dioxetane, 3-hydroxymethyl 3,4,4-trimethyl-1,2-dioxetane (HTMD), 3,3,4,4-tetramethyl-1,2-dioxetane and 3,4,4 trimethyl-1,2-dioxetane (TrMD), which on thermal decomposition generate triplet excited carbonyl products. To monitor quantitatively the formation of the mutagenic oxidation product 7,8-dihydro-8-oxoguanine (8-oxoGua), a sensitive and selective HPLC electrochemical assay was used after acidic hydrolysis (HF/pyridine) of the dioxetane-treated DNA. High yields of 8-oxoGua (up to ca 4% of the available guanine) were obtained for HTMD and TrMD. Both were investigated in detail with respect to effects of concentration, time and temperature. The oxidative reactivity of 1,2-dioxetanes was compared with several type I (benzophenone and riboflavin) and type II (methylene blue and rose bengal) photooxidants and disodium 1,4-etheno-2,3-benzodioxin-1,4-dipropionate as a chemical source of singlet oxygen. The persistence of 8-oxoGua towards oxidation by HTMD was examined in the reaction with 7,8-dihydro-8-oxo-2'-deoxyguanosine (8 oxodGuo) and with oxidized DNA. It was shown that, indeed, 8-oxoGua is consumed in the oxidized DNA on prolonged exposure to an excess of HTMD. The reaction of 8 oxodGuo with HTMD afforded the two 4R* and 4S* diastereomers of 9-(2-deoxy-beta-D erythropentofuranosyl)-4, 8-dihydro-4-hydroxy-8-oxoguanine as main oxidation products. Trapping experiments with tert-butanol confirmed that hydroxyl radicals are not involved, whereas the use of the triplet quenchers sodium 9,10-dibromo anthracene-2-sulfonate and 2,3-diazabicyclo[2.2.1]hept-2-ene established that triplet-excited states are mainly responsible for the observed DNA oxidation through type I action (electron transfer chemistry). The role of singlet oxygen was tested by means of deuterium isotope effects in D2O versus H2O, but no definitive conclusion could be reached in regard to the involvement of 1O2 in these oxidations.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7480133 TI - Spontaneous recovery of fluorescence by photobleached surface-adsorbed proteins. AB - Fluorescence photobleaching of a carboxyfluorescein-labeled protein (erythrocyte cytoskeletal protein 4.1) immobilized on bare glass is found to be spontaneously reversible, provided that the sample is deoxygenated. After a short (hundredths of seconds) photobleaching laser flash, the subsequent fluorescence excited by a dim probe beam partly recovers on a long (tenths of second) time scale, even in the absence of chemical exchange or diffusion processes. Neither the fraction of the fluorescence that bleaches reversibly nor its recovery rate is a strong function of fluorophore surface concentration. At a fixed surface concentration, the reversibly photobleached fraction and its recovery rate decreases with increasing duration or intensity of the bleaching flash. On the other hand, nondeoxygenated air-equilibrated samples exhibit almost total irreversible bleaching on this time scale. Quantitative fluorescence microscopy experiments occasionally require deoxygenation to avoid photochemical crosslinking or photobleaching or to enhance the triplet state population. The observation presented here indicate that fluorescence recovery after photobleaching (FRAP) experiments performed under deoxygenated conditions for measuring diffusion or chemical kinetics should be interpreted with caution: fluorescence recoveries may be due to intrinsic photochemical processes rather than fluorophore mobility. The recovery effect appears too slow to be ascribed simply to a relaxation of a triplet state; other possible explanations are offered. PMID- 7480134 TI - Comparison of the solid-matrix luminescence properties of benzo(a)pyrene-DNA adducts on alpha-cyclodextrin/NaCl and trehalose/NaCl matrices. AB - The solid-matrix luminescence properties of (+/-)-trans-7,8-dihydroxy-anti-9, 10 epoxy-7,8,9,10-tetrahydrobenzo(a)pyrene-DNA ([+/-]-anti-BPDE-DNA) adducts were compared on alpha-cyclodextrin (CD)/NaCl and trehalose/NaCl solid matrices. Both the optimum composition for the solid matrices and the best solvent system were obtained experimentally for acquiring the maximum room-temperature fluorescence (RTF) and room-temperature phosphorescence (RTP) signals for the (+/-)-anti-BPDE DNA. Most of the solid-matrix RTF and RTP data were obtained at 296 K and 93 K for (+/-)-anti-BPDE-DNA adducts adsorbed on 1% alpha-CD/NaCl and 80% trehalose/NaCl. The RTF signals were strong for (+/-)-BPDE-DNA adducts on both solid matrices, but RTP was only obtained on the trehalose/NaCl solid matrices with the 80% trehalose yielding the strongest RTP signal for (+/-)-anti-BPDE-DNA. The fluorescence lifetime data for (+/-)-anti-BPDE-DNA gave two components on 1% alpha-CD/NaCl. For 80% trehalose/NaCl, three components were revealed, but two components were obtained with 80% trehalose/NaCl after ether extraction of the solid matrix. The third component was ascribed to the formation of the tetrols from (+/-)-anti-BPDE-DNA adducts during the drying step in the sample preparation of 80% trehalose/NaCl. The results give the first reported data on the solid matrix luminescence of the (+/-)-anti-BPDE-DNA adducts. These results should be of considerable interest not only from an analytical viewpoint but as a new means of studying the luminescence characteristics of the adducts. PMID- 7480135 TI - Investigation of light-induced conformation changes in spiropyran-modified succinylated poly(L-lysine). AB - To determine the maximum range of coupling between side-chain photochromism and polypeptide conformation change, we modified the carboxylate side chains of succinylated poly(L-lysine) with a spiropyran to form polypeptide I. The extent of modification was determined to be 35.5%. The spacer group length between the polypeptide alpha-carbon and the dye was 12 atoms, providing minimum polypeptide dye interaction. Conformation changes were monitored by circular dichroism as a function of light adaptation and solvent composition (hexafluoroisopropanol [HFIP] vs trifluoroethanol [TFE]). Under all solvent compositions, the dark adapted dye was in the merocyanine form. Light adaptation by visible light converted the dye to the spiropyran form. When dissolved in TFE, I adopted a helical conformation insensitive to light adaptation. With increasing percentage HFIP, a solvent-induced helix-to-coil transition was observed around 80% (vol/vol) HFIP. At 100% HFIP, both light- and dark-adapted forms of I were in the coil state. Near the midpoint of the solvent-induced helix-to-coil transition, light adaptation caused conformation changes. Applying helix-to-coil transition theory, we measured a statistically significant difference in coil segment-HFIP binding constant for light- vs dark-adapted solutions (6.38 +/- 0.03 M-1 vs 6.56 +/- 0.03 M-1), but not for the nucleation parameter sigma (1.2 +/- 0.4 10(-3) vs 1.3 +/- 0.3 x 10(-3). The small binding constant difference translated to a light induced binding energy difference of 17 cal/mol/monomer. Near the midpoint of the helix-to-coil transition, collective interactions between monomer units made possible the translation of a small energy difference (less than RT) into large macromolecular conformation changes.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7480136 TI - Identification of a large genomic region in UV-irradiated human cells which has fewer cyclobutane pyrimidine dimers than most genomic regions. AB - Size separation after UV-endonuclease digestion of DNA from UV-irradiated human cells using denaturing conditions fractionates the genome based on cyclobutane pyrimidine dimer content. We have examined the largest molecules available (50-80 kb; about 5% of the DNA) after fractionation and those of average size (5-15 kb) for content of some specific genes. We find that the largest molecules are not a representative sampling of the genome. Three contiguous genes located in a G+C rich isochore (tyrosine hydroxylase, insulin, insulin-like growth factor II) have concentrations two to three times greater in the largest molecules. This shows that this genomic region has fewer pyrimidine dimers than most other genomic regions. In contrast, the beta-actin genomic region, which has a similar G+C content, has an equal concentration in both fractions as do the p53 and beta globin genomic regions, which are A+T-rich. These data show that DNA damage in the form of cyclobutane pyrimidine dimers occurs with different probabilities in specific isochores. Part of the reason may be the relative G+C content, but other factors must play a significant role. We also report that the transcriptionally inactive insulin region is repaired at the genome-overall rate in normal cells and is not repaired in xeroderma pigmentosum complementation group C cells. PMID- 7480138 TI - Effect of hematoporphyrin derivative on hematological parameters in rats. AB - Wistar rats were injected with hematoporphyrin derivative (Hpd) intraperitoneally and kept in the dark. Rats were sacrificed 2,24,48 and 72 h after injection. It was observed that Hpd in the dark did not affect the hemoglobin content and number of erythrocytes, while the leukocyte count was increased and blood pH decreased. Blood levels of glucose and lactate were increased significantly. Because the food intake was similar in all the groups, glycogenolysis was suspected to be the source of increased glucose levels in blood. However, a significant increase in the glycogen content of the livers of Hpd-treated rats was observed, which rules out glycogenolysis. Hyperglycemia may result due to a number of reasons such as stimulation of the central nervous pathways innervating the liver and adrenal medulla, excessive glucogenesis in liver from glycogen and noncarbohydrate sources, emotional stress, anesthesia and hormonal effects. The present study rules out hyperglycemia due to anesthesia and glucogenesis in the liver. Maintenance of blood glucose levels is a highly complex mechanism. Further investigations to understand these mechanisms are in progress. PMID- 7480137 TI - Cloning, expression and sequence analysis of cDNA for the luciferases from the Japanese fireflies, Pyrocoelia miyako and Hotaria parvula. AB - Cloning and sequence analysis of cDNA for the luciferases of Pyrocoelia miyako and Hotaria parvula were carried out (GenBank accession numbers L39928 and L39929, respectively). The amino acid sequence, deduced from the nucleotide sequence, showed P. miyako luciferase to consist of 548 amino acid residues with a molecular weight of 60,955, while the luciferase of H. parvula consisted of 548 amino acid residues with a molecular weight of 60,364. Pyrocoelia miyako luciferase showed 82.1% homology with the luciferase of Photinus pyralis and less than 70% homology with other firefly luciferases, whereas H. parvula luciferase showed 98%, 82.5% and 81.2% homology with the luciferases of Luciola mingrelica, Luciola lateralis and Luciola, cruciata respectively. Two regions in the enzymes were found to be highly conserved. The amino acid sequences were used to construct a phylogenetic tree, which showed that the fireflies could be divided into two groups. PMID- 7480140 TI - In vivo excitation of photosensitizers by infrared light. AB - Use of near infrared instead of visible light would markedly improve tissue penetration, making larger tumors candidates for photochemotherapy. Because common photesensitizers exhibit virtually no absorption in this wavelength region, conditions are required where the simultaneous action of two photons is possible. Healthy tissue (rat ears), sensitized by hematoporphyrin derivative, sulfonated chloroaluminum phthalocyanine or pheophorbide alpha, was irradiated (1064 nm, 10 ns) with power densities up to 200 MW cm-2 and total energy densities up to 200 kJ cm-2. No reproducible photodynamic lesions were observed, but there was sensitizer fluorescence that depended quadratically on the excitation intensity. PMID- 7480139 TI - The effect of chronic low-dose UVB radiation on Langerhans cells, sunburn cells, urocanic acid isomers, contact hypersensitivity and serum immunoglobulins in mice. AB - C3H mice were irradiated three times a week for up to 6 weeks with either 500 J/m2 or 1000 J/m2 broadband UVB (270-350 nm) or 3000 J/m2 narrowband UVB (311-312 nm; TL01 source). Each dose was suberythemal to the mouse strain used. The number of Langerhans cells (LC) in the epidermis was reduced by over 50% after 2 weeks of irradiation with the UVB source and by 20% following TL01 irradiation. Continued irradiation for up to 6 weeks resulted in no further decrease in LC numbers in the case of the UVB source but a steady decline to 40% in the case of the TL01 source. Sunburn cells were detected following irradiation with both sources but the numbers were very low in comparison with acute exposure. Ultraviolet-B exposure resulted in doubling of the thickness of the epidermis throughout the 6 weeks of irradiation while TL01 exposure did not alter epidermal thickness. Conversion of trans- to cis-urocanic acid (UCA) was observed with both UVB and TL01 sources. The percentage of cis-UCA started to return to normal after 4 weeks of TL01 exposure despite continued irradiation. As observed following a single exposure, the contact hypersensitivity (CH) response was significantly reduced following 6 weeks of UVB irradiation but was unaffected by TL01 exposure, indicating no correlation between cis-UCA levels and CH response. Total serum immunoglobulin levels remained unchanged throughout the 6 weeks of UVB or TL01 irradiation but IgE titers significantly increased in all cases in the first 2 weeks of irradiation, indicating a possible shift to a TH2 cytokine profile.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7480141 TI - Age-related changes in the human lens as monitored by detection of porphyrin excited states. AB - Previous studies have shown that the triplet state lifetimes of various porphyrins are increased by several orders of magnitude when they are bound to lens protein. Flash photolysis studies of mesotetra (p-sulfonatophenyl)porphyrin (TPPS) on intact bovine lenses indicated a biexponential decay of the triplet state with lifetimes of 160 microsecond and 1.6 ms. Here we extend those measurements to TPPS associated with intact human lenses. Steady-state fluorescence measurements indicate that TPPS binds to both young and old human lenses. In an intact young human lens, the TPPS triplet state is observed to decay biexponentially with lifetimes of 50 and 680 microsecond. As the age of the lens increases, the lifetime of the shorter-lived component lengthens while that of the longer-lived component decreases slightly. In order human lenses, the two lifetimes coalesce and the triplet decay exhibits purely monoexponential behavior. These photophysical characteristics apparently are due to age-related modification(s) of the protein in the human lens resulting in an increasingly more homogeneous environment around the porphyrin. PMID- 7480142 TI - Structure-activity relationship of porphines for photoinactivation of bacteria. AB - The antibacterial photodynamic effects of uncharged (o-tetrahydroxyphenyl porphine [THPP], m-THPP and p-THPP), cationic (5,10,15,20-tetra[4-N methylpyridyl]porphine [TMPyP]) and anionic (5,10,15,20-tetra[4-sulfonatophenyl porphine] [TPPS4]) porphines on Staphylococcus aureus and Escherichia coli bacteria inactivation were examined. The results show that uncharged porphines provoked antibacterial photodynamic activity on S. aureus, and also on E. coli in the presence of the membrane-disorganizing peptide polymixin B nonapeptide (PMNP). The TMPyP compound was highly photoactive toward gram-positive bacteria but only marginally effective on gram-negative cells, whereas TPPS4 showed no activity on either gram-positive or gram-negative bacteria. The photoactivity of TMPyP is due to the electrostatic attraction between the positively charged sensitizer molecule and the negatively charged membrane of the gram-positive target cells. For TPPS4, the inactivity toward gram-positive bacteria is due to electrostatic repulsion between the charged sensitizer molecule and the cell membrane. For gram-negative bacteria, the inactivity is conceivably due to preferential (electrostatic) binding to the positively charged PMNP, which is an adjuvant for membrane disorganization, but has no effect on cell viability. For hydrophobic sensitizers, the photoactivity depends on the state of aggregation. The extent of deaggregation of the different THPP isomers was determined by fluorescence measurements of bound sensitizers and could be positively correlated with their photoinactivation capacity. We conclude that the structure-activity relationships of these porphines are affected by their net charge and by aggregation. PMID- 7480143 TI - Quantitative assessment of cumulative damage from repetitive exposures to suberythemogenic doses of UVA in human skin. AB - Daily exposures to relatively small suberythemogenic fluences of UVA (50-200 kJ/m2) for 8 days resulted in cumulative morphological skin alterations indicative of early tissue injury. Histologically, irradiated skin revealed epidermal hyperplasia, inflammation and deposition of lysozyme along the dermal elastic fiber network. Sunburn cells were also present within the epidermis. These changes were quantified by image analysis and were found to be related to the cumulative UVA fluence. A long UVA waveband (UVAI, 340-400 nm) was as effective as a broad UVA band (320-400 nm), suggesting that these changes are induced by longer UVA wavelengths. PMID- 7480144 TI - Alterations in erythrocyte band 3 organization induced by the photosensitizer, hematoporphyrin derivative. AB - Photosensitization of erythrocytes in the presence of hematoporphyrin derivative causes cross-linking of membrane proteins. This cross-linking is associated with partial lysis of the cells and an increased susceptibility to heat-induced membrane fragmentation. The effect of photosensitization on the organization of erythrocyte band 3 was monitored using the technique of time-resolved phosphorescence anisotropy. Band 3 rotational diffusion was somewhat restricted upon photooxidation, indicating aggregation of this major integral membrane protein. PMID- 7480145 TI - Low-temperature trapping of early photointermediates of alpha-isorhodopsin. AB - Alpha-Isorhodopsin, an artificial visual pigment with a 9-cis-4,5-dehydro-5,6 dihydro(alpha)retinal chromophore, was photolyzed at low temperatures and absorption difference spectra were collected as the sample was warmed. A bathorhodopsin (Batho)-like intermediate absorbing at ca 495 nm was detected below 55 K,a blue-shifted intermediate (BSI)-like intermediate absorbing at ca 453 nm was observed when the temperature was raised to 60 K and a lumirhodopsin (Lumi)-like intermediate absorbing at ca 470 nm was found when the sample was warmed to 115 K. Photointermediates from this pigment were compared to those of native rhodopsin and 5,6-dihydroisorhodopsin. As in native rhodopsin, Batho is the first intermediate detected in alpha-isorhodopsin, though unlike native rhodopsin at low temperatures BSI is observed prior to Lumi formation. Alpha Isohodopsin behaves similarly to 5,6-dihydroisorhodopsin, with the same early intermediates observed in both artificial visual pigments lacking the C5-C6 double bond. The transition temperature for BSI formation is higher in alpha isorhodopsin, suggesting an interaction involving the chromophore ring in BSI formation. The transition temperature for Lumi formation is similar for these two pigments as well as for native rhodopsin, suggesting comparable changes in the protein environment in that transition. PMID- 7480146 TI - Photostationary state compositions of retinal and related compounds included in beta-lactoglobulin. Effects of protein host on isomer distribution of polyene substrates. AB - The UV-visible absorption spectra and photostationary state compositions of retinal (or the related C-18 ketone, 3-dehydroretinal and the C-22 aldehyde) imbedded in the binding cavity of beta-lactoglobulin (BLG) are consistent with the view that the carbonyl group of these polyenes are hydrogen-bonded with the protein host, most likely with the lone protonated lysine residue in the binding pocket. Patterns of variation in photochemical behavior of the imbedded chromophore versus that in solution are discussed in terms of possible specific protein-substrate interactions. The results are also compared with that of the methyl ether of retinol where similar hydrogen bonding is not possible. PMID- 7480147 TI - Chemical aspects of bioluminescence. PMID- 7480148 TI - Biochemistry of bacterial bioluminescence. PMID- 7480149 TI - Green fluorescent protein. AB - Several bioluminescent coelenterates use a secondary fluorescent protein, the green fluorescent protein (GFP), in an energy transfer reaction to produce green light. The most studied of these proteins have been the GFPs from the jellyfish Aequorea victoria and the sea pansy Renilla reniformis. Although the proteins from these organisms are not identical, they are thought to have the same chromophore, which is derived from the primary amino acid sequence of GFP. The differences are thought to be due to changes in the protein environment of the chromophore. Recent interest in these molecules has arisen from the cloning of the Aequorea gfp cDNA and the demonstration that its expression in the absence of other Aequorea proteins results in a fluorescent product. This demonstration indicated that GFP could be used as a marker of gene expression and protein localization in living and fixed tissues. Bacterial, plant and animal (including mammalian) cells all express GFP. The heterologous expression of the gfp cDNA has also meant that it could be mutated to produce proteins with different fluorescent properties. Variants with more intense fluorescence or alterations in the excitation and emission spectra have been produced. PMID- 7480150 TI - Molecular evolution of the Ca(2+)-binding photoproteins of the Hydrozoa. AB - Alignment of the primary structures of the hydrozoan photoproteins, aequorin, mitrocomin, clytin and obelin showed very strong amino acid sequence identities. The Ca(2+)-binding sites of the proteins were found to be highly conserved. The Ca(2+)-binding sites were also homologous to the Ca(2+)-binding sites of other Ca(2+)-binding proteins. However, aequorin, mitrocomin, clytin and obelin differed from other Ca(2+)-binding proteins in that they contained a relatively large number of cysteine, tryptophan, histidine, proline and tyrosine residues, suggesting that these residues may have evolved as part of the light-emitting mechanism. Construction of a phylogenetic tree showed that aequorin, mitrocomin, clytin and obelin form a closely related group of proteins. PMID- 7480151 TI - Interaction of photodynamically induced cell killing and dark cytotoxicity of rhodamine 123. AB - Loss of clonogenicity of Chinese hamster ovary (CHO) cells, murine L929 fibroblasts and human bladder carcinoma T24 cells caused by photodynamic treatment (PDT) with hematoporphyrin derivative (HPD) is synergistically enhanced by subsequent incubation with rhodamine 123 in the dark. For CHO and L929 cells this synergistic interaction can be explained by an increased uptake of rhodamine 123 as the result of the photodynamic treatment. With aluminum phthalocyanine (AIPC) as photosensitizer only additive effects were observed in the three cell lines. Incubation in the dark with rhodamine 123, followed by a photodynamic treatment with HPD, resulted in an antagonistic interaction with regard to loss of colony formation. With AIPc the combination of treatments resulted in an additive effect with L929 and T24 cells, whereas with CHO cells a slight antagonistic interaction was observed. An antagonistic effect was also observed in model experiments, treating histidine photodynamically with HPD and measuring oxygen consumption. A possible explanation of these results could be an interaction or complex formation of rhodamine 123 with HPD resulting in a diminished singlet oxygen production. With AIPc this does not take place. PMID- 7480152 TI - Evaluation of new benzoporphyrin derivatives with enhanced PDT efficacy. AB - A first report on the biological evaluation of a series of isomerically pure benzoporphyrin derivatives (cis- and trans-isomers) as methyl esters is described. In preliminary in vivo studies, the n-hexyl ether analogues of both cis- and trans-isomers of benzoporphyrin derivatives were found to be more active than the industrially prepared benzoporphyrin derivative, a mixture of monocarboxylic acids (BPDMA, Quadralogic Technologies, Vancouver). Further studies with 4-de-vinyl-4- (1-hexyloxyethyl) benzoporphyrin derivative showed that, like BPDMA, it had reduced residual skin phototoxicity compared in mice with Photofrin. The uptake and clearance characteristics of BPDMA were also compared with the 4-(1-hexyloxyethyl)-derivative by in vivo reflection spectroscopy. PMID- 7480153 TI - Ultraviolet erythema sensitivity in anamnestic (I-IV) and phototested (1-4) Caucasian skin phototypes: the need for a new classification system. AB - The anamnestic skin phototypes (ASP) I-IV of 22 Caucasian volunteers wee compared with their phototested skin phototypes (PSP) using solar simulating, broadband UV radiation. The Commission Internationale de l'Eclairage (CIE)-weighted (i.e. erythemally effective) minimal erythema doses (MED) for solar simulating radiation varied from 20 mJ/cm2 (PSP type 1) to 57 mJ/cm2 (PSP type 4). In only 11 of 21 volunteers did the ASP (I-IV) and PSP (1-4) classifications coincide, and the MED values of the volunteers within the different ASP groups (I-IV) overlapped considerably. To compare the reactivity to erythematogenic radiation of different wavelengths, narrowband monochromator irradiations were performed at 298 nm, 310 nm and 330 nm. The CIE-weighted MED values at these wavelengths (20 80 mJ/cm2) corresponded well with those obtained in the broadband testing. Our results indicate that, with classification by interrogation, Caucasian skin can reliably be classified into only two subtypes, corresponding to Fitzpatrick phototypes I-III and phototype IV, respectively. A classification into four sensitivity types can be achieved by phototesting, only. We propose that the concept of ASP should be used with caution. The concept of PSP 1-4 should be favored. PMID- 7480154 TI - Carcinogenic and melanogenic effects of a filtered metal halide UVA source and a tubular fluorescent UVA tanning source with or without additional solar-simulated UV radiation in hairless mice. AB - The carcinogenic and melanogenic effects of a filtered metal halide source (UVASUN) that emits UV radiation in a range from 340 to 400 nm and a bank of Philips TL 09R tubes (TL 09) emitting in a range from 310 to 400 nm were studied in lightly pigmented hairless hr/hr C3H/Tif mice. Both the carcinogenic effect of the two UVA radiation sources alone and in combination with a UV source, consisting of one Philips TL 12 and five Bellarium-S SA-1-12 tubes emitting radiation somewhat similar to the UV part of the solar spectrum (SOLAR UV), were investigated. Finally, the melanogenic effect of exposure to the two UVA sources were studied. The mice were exposed to the UVA sources 30 min/day 5 days/week, in equal erythemogenic doses, calculated by using the Commission Internationale de l'Eclairage human erythema action spectrum. Equal erythemogenic doses of TL 09 and UVASUN induced the same degree of skin pigmentation, but skin tumor development was enhanced in mice exposed to TL 09 compared with UVASUN (P < 0.0005). For all but one tumor, endpoint pretreatment with TL 09 or UVASUN for 91 days did not influence tumor development during subsequent exposure to SOLAR UV radiation 10 min/day, 4 days/week. Exposure to the two UVA radiation sources after 91 days of SOLAR UV exposure significantly enhanced skin tumor development. Overall, the data on the interaction between exposure to the UVA sources and SOLAR UV indicated that the risk of SOLAR UV-induced carcinogenesis was independent of the type of prior-UVA exposure and post-UVA exposure. PMID- 7480155 TI - Photosensitization of murine tumor, vasculature and skin by 5-aminolevulinic acid induced porphyrin. AB - The effects of topical and systemic administration of 5-aminolevulinic acid (ALA) were examined in several murine tumor systems with regard to porphyrin accumulation kinetics in tumor, skin and blood, vascular and tumor cell photosensitization and tumor response after light exposure. Marked, transient increases in porphyrin levels were observed in tumor and skin after systemic and topical ALA. Rapid, transient, dose-dependent porphyrin increases were also observed in blood; these were pronounced after systemic ALA injection and mild after topical application. They were highest within 1 h after ALA injection, thereafter declining rapidly. This matched the clearing kinetics of injected exogenous protoporphyrin IX (PpIX). Initially, vascular photosensitivity changed inversely to blood porphyrin levels, increasing gradually up to 5 h post-ALA, as porphyrin was clearing from the bloodstream. This pattern was again matched by injected, exogenous PpIX. After therapeutic tumor treatment vascular disruption of the tumor bed, while observed, was incomplete, especially at the tumor base. Minimal direct tumor cell kill was found at low photodynamic therapy (PDT) doses (250 mg/kg ALA, 135 J/cm2 light). Significant, but limited (< 1 log) direct photodynamic tumor cell kill was obtained when the PDT dose was raised to 500 mg/kg systemic ALA, followed 3 h later by 270 J/cm2, a dose that was however toxic to the animals. The further reduction of clonogenic tumor cells over 24 h following treatment was moderate and probably limited by the incomplete disruption of the vasculature. Tumor responses were highest when light treatment was carried out at the time of highest tumor porphyrin content rather than at the time of highest vascular photosensitivity. Tumor destruction did not reach the tumor base, regardless of treatment conditions. PMID- 7480157 TI - On the possibility of calcium imaging using Indo-1 with three-photon excitation. AB - We show that the calcium fluorophore Indo-1 can be excited by simultaneous absorption of three-photons at 885 nm, a wavelength readily available from Ti:sapphire lasers. Three-photon excitation was demonstrated by the emission intensity of Indo-1 which depended on the cube of the laser power, and by a higher anisotropy than was observed for two-photon excitation. Excitation of Indo 1 becomes a two-photon process when the wavelength is decreased to 820 nm. Three photon excitation was accomplished at a low 17 microM concentration of Indo-1. Examination of the spatial profile of the excited Indo-1 showed a smaller volume for three- versus two-photon excitation. These results suggest that three-photon excitation may be useful in fluorescence microscopy using the long wavelength output of Ti:sapphire lasers, and may provide higher spatial resolution than available using two-photon excitation. PMID- 7480156 TI - Potentiation of merocyanine 540-mediated photodynamic therapy by salicylate and related drugs. AB - Simultaneous exposure to merocyanine 540 (MC540) and light of a suitable wavelength kills leukemia, lymphoma and neuroblastoma cells but is relatively well tolerated by normal pluripotent hematopoietic stem cells. This differential phototoxic effect has been exploited in preclinical models and a phase I clinical trial for the extracorporeal purging of autologous bone marrow grafts. Salicylate is known to potentiate the MC540-mediated photokilling of tumor cells. Assuming that salicylate induces a change in the plasma membrane of tumor cells (but not normal hematopoietic stem cells) that enhances the binding of dye molecules it has been suggested that salicylate may provide a simple and effective means of improving the therapeutic index of MC540-mediated photodynamic therapy. We report here on a direct test of this hypothesis in a murine model of bone marrow transplantation as well as in clonal cultures of normal murine hematopoietic progenitor cells. In both systems, salicylate enhanced the MC540-sensitized photoinactivation of leukemia cells and normal bone marrow cells to a similar extent and thus failed to improve the therapeutic index of MC540 significantly. On the basis of a series of dye-binding studies, we offer an alternative explanation for the potentiating effect of salicylate. Rather than invoking a salicylate-induced change in the plasma membrane of tumor cells, we propose that salicylate displaces dye molecules from serum albumin, thereby enhancing the concentration of free (active) dye available for binding to tumor as well as normal hematopoietic stem cells. PMID- 7480158 TI - Insights into cellular energy metabolism from transgenic mice. AB - Rapid progress in manipulating the mouse genome now makes it possible to express any cloned gene in virtually any cell type of the mouse. Homologous recombination in embryonic stem cells allows the specific mutation of any gene. These tools are finding widespread application to problems in cell biology and physiology. After a brief description of some of the molecular genetic techniques available for the mouse, the application of transgenic mice to problems in cellular energy metabolism is discussed. The effects of increasing glucose transport on carbohydrate metabolism in muscle and fat are presented. Experiments using a transgenic mouse expressing creatine kinase in liver to understand ATP catabolism and regulation of oxidative phosphorylation are discussed. Finally, mice either lacking or misexpressing different isoforms of creatine kinase have been useful for understanding the detailed role of this important enzyme in cellular energy metabolism. PMID- 7480159 TI - Nonsynaptic modulation of neuronal activity in the brain: electric currents and extracellular ions. AB - Nonsynaptic interactions between neurons have been eclipsed by our increasingly detailed understanding of chemical synapses, but they do play significant roles in the nervous system. This review considers four classes of nonsynaptic interaction, mainly in mammalian brain. 1) Electrotonic (and chemical) coupling through gap junctions has effects during development and under some, often pathological, conditions in the mature brain. 2) Ephaptic transmission is mediated by electrical coupling between specific neuronal elements in the absence of specialized contacts, notably in the cerebellum, and in axon tracts affected by demyelination. 3) Field effect interactions are mediated by large extracellular currents and potential fields generated by the hippocampus and other cortical structures. Both endogenous and applied electric fields alter neuronal excitability at field strengths over a few millivolts per millimeter. Weaker fields have more subtle effects, for instance, on axonal growth during development and repair and, more controversially, in behavioral responses to environmental fields. 4) There are fluctuations in extracellular ions such as K+, which are released during neuronal activity and which alter neuronal excitability. Field effects and ion fluctuations probably have modest effects during physiological activity but have a significant impact on epileptic seizures, and can sustain them in the absence of synaptic transmission. PMID- 7480160 TI - Cyclic nucleotide phosphodiesterases: functional implications of multiple isoforms. AB - In the last few years there has been a veritable explosion of knowledge about cyclic nucleotide phosphodiesterases. In particular, the accumulating data showing that there are a large number of different phosphodiesterase isozymes have triggered an equally large increase in interest about these enzymes. At least seven different gene families of cyclic nucleotide phosphodiesterase are currently known to exist in mammalian tissues. Most families contain several distinct genes, and many of these genes are expressed in different tissues as functionally unique alternative splice variants. This article reviews many of the more important aspects about the structure, cellular localization, and regulation of each family of phosphodiesterases. Particular emphasis is placed on new information obtained in the last few years about how differential expression and regulation of individual phosphodiesterase isozymes relate to their function(s) in the body. A substantial discussion of the currently accepted nomenclature is also included. Finally, a brief discussion is included about how the differences among distinct phosphodiesterase isozymes are beginning to be used as the basis for developing therapeutic agents. PMID- 7480161 TI - Human surfactant protein B: structure, function, regulation, and genetic disease. AB - Elucidation of the structure and function of the hydrophobic surfactant protein (SP-B) and the SP-B gene has provided critical insight into surfactant homeostasis and control of respiratory epithelial cell gene expression. Surfactant protein B, in concert with surfactant protein A (SP-A), surfactant protein C (SP-C), and surfactant phospholipids, contributes to the structure and function of surfactant particles, determining surface activities and pathways by which surfactant phospholipids and proteins are processed, routed, packaged, and secreted from lamellar bodies by type II epithelial cells. After secretion, SP-B plays an essential role in determining the structure of tubular myelin, the stability and rapidity of spreading, and the recycling of surfactant phospholipids. The biochemical and structural signals underlying the homeostasis of alveolar surfactant are likely mediated by interactions between the surfactant proteins and phospholipids producing discrete structural forms that vary in size, aproprotein, and phospholipid content. Distinctions in structure, protein, and size are likely to determine the function of surfactant particles, their catabolism, or recycling by alveolar macrophages and airway epithelial cells. Analysis of the genetic controls governing the SP-B gene has led to the definition of DNA-protein interactions that determine respiratory epithelial cell gene expression in general. The important role of SP-B in lung function was defined by the study of a lethal neonatal respiratory disease, hereditary SP-B deficiency, caused by mutations in the human SP-B gene. PMID- 7480162 TI - Anatomy and electrophysiology of fast central synapses lead to a structural model for long-term potentiation. AB - Detailed knowledge of the anatomy of central synapses is essential to the interpretation of the vast quantity of electrophysiological findings that have been published in recent years. When their function is considered, it is not surprising that, in both anatomy and electrophysiology, fast central synapses show important differences to the neuromuscular junction. This review concentrates on the detailed anatomy of the common excitatory synapses that impinge on dendritic spines, but also refers to other glutamatergic and GABAergic synapses. This information is brought together with present knowledge of the electrophysiology of fast neurotransmission in the brain. Various types of evidence are outlined, explaining why it is now widely accepted that release of transmitter from a single vesicle virtually saturates the small number of receptors available on the postsynaptic membrane of central synapses. Finally, the anatomic literature suggests that a particular type of spine synapse, which electron microscopy reveals to have a perforated active zone, may represent a synapse with high efficacy. This suggestion is shown to be completely compatible with the electrophysiological data, and a model is presented that shows that all the apparently conflicting data in the field of long-term potentiation could be compatible. This stresses the need for cooperative collaboration between laboratories that have apparently conflicting findings. PMID- 7480163 TI - Dynamics of nerve-muscle interaction in developing and mature neuromuscular junctions. AB - Neuromuscular connections have long served as models of synaptic structure and function. They also provide illuminating insights into the dynamic cell-cell interactions governing synaptogenesis, neuromuscular differentiation, and the maintenance of effective function. This paper reviews recent advances in our understanding of the regulatory and inductive interactions involved in motor axon pathfinding, target recognition, bidirectional control of gene expression during synapse formation, motoneuron cell death, terminal rearrangement, and the ongoing remodeling of synaptic number, structure, and function to adjust to growth and changes in use. PMID- 7480164 TI - Role of microtubule-associated proteins in the control of microtubule assembly. AB - In eukaryotic cells, microtubules, actin, and intermediate filaments interact to form the cytoskeletal network involved in determination of cell architecture, intracellular transport, modulation of surface receptors, mitosis, cell motility, and differentiation. Cytoskeletal organization and dynamics depend on protein self-associations and interactions with regulatory elements such as microtubule associated proteins (MAPs). The MAP family includes large proteins like MAP-1A, MAP-1B, MAP-1C, MAP-2, and MAP-4 and smaller components like tau and MAP-2C. This review focuses on relevant aspects of MAP function, with emphasis on their roles in modulating cytoskeletal interactions. In this context, MAP expression mechanisms and posttranslational modifications are also discussed. Microtubule associated proteins have a rather widespread distribution among cells, but certain MAPs have been identified in specific cell types. Within single neurons, MAP-2 is dendritic while tau is preferentially an axonal protein. Their expression is developmentally regulated. Even though MAPs share a capacity to interact with the COOH-terminal tubulin domain, stabilize microtubules, and link them with other cytoskeletal polymers, they exhibit structural differences. However, MAP-2, MAP-4, and tau have common repetitive microtubule-binding motifs. Microtubule-associated proteins not only control cytoskeletal integrity, but they also appear to interact with highly structural elements of cells. Molecular biological approaches permitted localization of new MAPs in cultured mammalian cells and invertebrate organisms and other microtubule-interacting proteins that exhibit transient interactions with microtubules. The structural/functional aspects of several new MAP-like proteins in centrosomes and the mitotic spindle, functionally implicated in cell cycle events, are also analyzed. PMID- 7480165 TI - Ion channel regulation by G proteins. AB - Ion channels are poised uniquely to initiate, mediate, or regulate such distinct cellular activities as action potential propagation, secretion, and gene transcription. In retrospect, it is not surprising that studies of ion channels have revealed considerable diversities in their primary structures, regulation, and expression. From a functional standpoint, the various mechanisms coopted by cells to regulate channel activity are particularly fascinating. Extracellular ligands, membrane potential, phosphorylation, ions themselves, and diffusible second messengers are all well-established regulators of ion channel activity. Heterotrimeric GTP-binding proteins (G proteins) mediate many of these types of ion channel regulation by stimulating or inhibiting phosphorylation pathways, initiating intracellular cascades leading to elevation of cytosolic Ca2+ or adenosine 3',5'-cyclic monophosphate levels, or by generating various lipid derived compounds. In some cases, it seems that activated G protein subunits can interact directly with ion channels to elicit regulation. Although there is currently little direct biochemical evidence to support such a mechanism, it is the working hypothesis for the most-studied G protein-regulated ion channels. PMID- 7480166 TI - Ligation of nonmatching DNA molecule ends. AB - T4 DNA ligase can promote the in vitro ligation of blunt DNA ends to ends bearing a 2-nucleotide single-stranded protrusion. This was shown by digestion of plasmids pBR322 and pSP71 with the appropriate restriction enzymes followed by recircularization of the plasmids and transformation of Escherichia coli. It could be ruled out that such nonmatching ligations are due to the presence of contaminating nucleases. The efficiency of ligation is of the same order of magnitude as that obtained with blunt end ligations. The interaction of a number of different combinations of blunt and sticky ends, the latter bearing both 3' and 5' protrusions, was investigated. Ligation of nonmatching ends was shown to take place in all cases. Several ligation junctions were sequenced, showing that during the ligation process the 2-nucleotide protrusion is trimmed away. In two instances the ligation event was accompanied by the specific loss of either 3 or 15 nucleotide pairs as well as the protrusion. An intermolecular ligation involving nonmatching ends was also performed, demonstrating that this form of ligation can be usefully employed in molecular cloning experiments. PMID- 7480167 TI - Isolation and characterization of a novel insertion sequence element, IS1248, in Paracoccus denitrificans. AB - A new suicide vector, pRVS3, was constructed to facilitate gene replacements in the genome of Paracoccus denitrificans. In control experiments, incorporation of this suicide vector into the genome did not depend on the presence of homologous DNA. Using appropriate restriction enzymes, the suicide vector and flanking DNA were recovered from the genomic DNA. Sequence analysis demonstrated that both up- and downstream of the ex-integrant vector there was an element that showed high homology with bacterial insertion sequences (IS). Southern blot analysis of wild type and integrant strains revealed that at least four copies of this IS element reside in the P. denitrificans genome, one of which, designated IS1248, had been involved in the transpositional event described here. IS1248 is 830 bp long, has 13-bp imperfect inverted repeats at the borders, and contains five open reading frames. With respect to the organization and primary sequences of the open reading frames, IS1248 closely resembles IS869 and IS427 of Agrobacterium tumefaciens, IS402 of Pseudomonas cepacia, and ISmyco found in Mycobacterium tuberculosis. PMID- 7480168 TI - Characterization of a 3.5-kbp plasmid from Helicobacter pylori. AB - A 3.5-kbp plasmid (pHPM180) was isolated from Helicobacter pylori (HPM180) and the DNA sequence was determined. Two open reading frames (ORF1 and ORF2) were identified which could encode polypeptides of 54,517 and 27,629 Da, respectively. Ribosome binding and promoter consensus sequences were identified, as well as two 232-bp direct repeats and four 22-bp direct repeats. DNA sequence homology was found between pHPM180 ORF1 and a 684-base pair HindIII fragment from a 5.6-kbp H. pylori plasmid. ORF1 showed amino acid homology with six replication proteins from bacterial plasmids with theta-type replicons. Additional sequence identity was found between pHPM180 noncoding DNA and a segment of H. pylori pHPK255, a plasmid that replicates via a rolling circle type mechanism. A ribonuclease protection assay determined that ORF1 was transcribed in H. pylori HPM180. PMID- 7480169 TI - Mutational analysis of SpvR binding to DNA in the regulation of the Salmonella plasmid virulence operon. AB - The Salmonella plasmid-borne spvR gene encodes a 33-kDa regulatory protein that activates transcription of the spvABCD operon during the stationary phase of bacterial growth. We used gel mobility shift assays to demonstrate that SpvR recognizes a specific target DNA sequence within a 318-bp EcoRI-ApaI fragment upstream of spvA. The addition of unlabeled target DNA to the radioactive labeled DNA-SpvR complex resulted in competitive inhibition of band retardation confirming the specificity of SpvR binding. Introduction of target DNA on a high copy number plasmid into wild-type Salmonella dublin Lane resulted in a substantial decrease of SpvB synthesis, confirming the binding properties of this DNA segment in vivo. Three SpvR mutants were constructed and were shown to abolish the positive regulatory function of SpvR. By site-specific mutagenesis of spvR, three single amino acids within the putative SpvR N-terminal alpha-helix domains were substituted by prolines. This resulted in loss of binding to the spvA promoter sequence and in loss of activation of the spvABCD genes. This study demonstrates that the regulatory function of SpvR is mediated by specific binding to the promoter region of the spvABCD operon. PMID- 7480170 TI - Dosage of Tn916 circular intermediates in Enterococcus faecalis. AB - Upon excision, conjugative transposon Tn916 forms a nonreplicating circular intermediate (CI), which is essential both for transposition and conjugal transfer. In this work we developed an assay to quantify the circular forms of Tn916. By this method we defined a new measurable trait of Tn916-carrying strains: the "CI copy number." CI dosage was performed by nested PCR (with primers designed on Tn916 termini) in a limiting dilution assay, where Poisson statistics were used to calculate the number of PCR amplification targets from the proportion of the negative endpoints. The number of CI was normalized to the number of bacterial chromosomes. This method enabled us to study the relationship between CI copy number, presence of tetracycline (the resistance marker of Tn916) in the culture medium, and conjugation frequency. Three isogenic strains of Enterococcus faecalis OG1 with Tn916 inserted at different sites on the chromosome were investigated for CI content. CI copy number varied depending on the strain, ranging between 7.8 and 610 copies per 10(6) chromosomes. Growth in liquid media containing tetracycline provoked an important increase both in CI copy number and conjugation frequency. This effect was more marked in low frequency donors. While cell-cell contact during filter mating did not produce an increase in CI copy number, Tn916 conjugation frequency was found to be dependent on CI copy number in donor cells. The dose-response curve showed a linear relationship with a slope of 0.74, for the entire range of conjugation frequencies tested (from 5.1 x 10(-8) to 2.8 x 10(-6) transconjugants per donor. PMID- 7480171 TI - Analysis of a retron EC86 and EC67 insertion site in Escherichia coli. AB - The Escherichia coli clinical strain 49 contains the integration sites for the retrons EC86 and EC67 but not the retrons themselves. To compare the chromosomal structures before and after retron integration, the DNA sequence of the integration site in strain 49 was determined and compared to the corresponding sequences in strains containing EC86 and EC67. The results suggested that when these retrons inserted into the E. coli chromosome they replaced a 3.5-kb fragment of chromosomal DNA. It is proposed that the replacement of preexisting DNA by a retron may be a general mechanism for retron integration, since in the three examples in which the integration sites are known, the retrons appear to have integrated into the chromosome by replacing a preexisting DNA segment. PMID- 7480172 TI - Multidrug resistance plasmid pSK108 from coagulase-negative staphylococci; relationships to Staphylococcus aureus qacC plasmids. AB - The 2.4-kb Staphylococcus epidermidis plasmid, pSK108, encodes a qacC multidrug resistance determinant. Sequence analysis has revealed that pSK108 is a member of the pC194 family of rolling circle replicating plasmids and suggests that the DNA segment containing qacC, which is bounded by the replication nick site and the minus origin palA, represents a resistance gene cassette that has undergone horizontal genetic exchange. PMID- 7480173 TI - Brazilin stimulates the glucose transport in 3T3-L1 cells. AB - Brazilin (7,11b-dihydrobenz[b]indeno-[1,2-d]pyran-3,6a,9,10(6H)- tetrol) was found to have hypoglycemic action and increase glucose metabolism in experimental diabetic animals. In order to investigate the mechanism of hypoglycemic action of brazilin, the effects of brazilin on glucose transport, insulin receptor autophosphorylation, and protein kinase C(PKC) activity in 3T3-L1 cells were studied. Brazilin increased basal glucose transport in 3T3-L1 fibroblasts and adipocytes. However, insulin-stimulated glucose transport was not influenced. Autophosphorylation of the partially purified insulin receptor was not affected by brazilin treatment in 3T3-L1 fibroblasts. However, brazilin decreased the PKC activity in 3T3-L1 fibroblasts and adipocytes. PMID- 7480174 TI - Effect of quercitrin on lactose-induced chronic diarrhoea in rats. AB - Quercitrin (3-rhamnosylquercetin) is a bioflavonoid contained in several crude drugs traditionally used for its antidiarrhoeal activity. The antidiarrhoeic effect of quercitrin on experimental chronic diarrhoea in rats was studied. Adult rats were fed for 14 days with a synthetic diet in which all soluble carbohydrates were substituted by lactose, resulting in chronic diarrhoea with body weight loss, colonic hyperplasia, reduced average cell size, increased alkaline phosphatase activity, increased mucus production and cytopathological alterations of the enterocyte. The rest of the animals were allowed to recover from chronic diarrhoea for 3 or 7 days, by feeding them with a standard diet, and half of them were also given quercitrin orally (50 mg/kg day). Diarrhoea ceased 48 h after lactose withdrawal, and body weight recovery was apparent after 3 days. Nevertheless, most of the alterations of the colonic mucosa persisted at that time. Quercitrin-treated rats had less diarrhoeal output and did not show mucosal hyperplasia after three days of treatment. All animals had greatly recovered by the seventh day, but histological alterations were still present, although to a lesser extent in quercitrin-treated rats. Quercitrin and related flavonoids may play a role in intestinal repair following chronic mucosal injury. PMID- 7480175 TI - Potent antiplatelet, anti-inflammatory and antiallergic isoflavanquinones from the roots of Abrus precatorius. AB - Five isoflavanquinones have been isolated from the roots of Abrus precatorius L. (Leguminosae). Three of them are new and designated as abruquinones D, E, and F. The pharmacological activities of the isoflavanquinones have been evaluated. The results indicated that abruquinones A, B, and D exhibited remarkable inhibitory effects on the platelet aggregation. The IC50 of abruquinones A and B for the inhibition of the platelet aggregation induced by arachidonic acid (AA) and collagen were less than 5 micrograms/ml, and of abruquinone D, was less than 10 micrograms/ml for that induced by AA. On the other hand, abruquinones A, B, D, and F showed strong anti-inflammatory and antiallergic effects. The IC50 of abruquinones A, B, D, and F for the inhibition of superoxide formation were less than 0.3 micrograms/ml, for the inhibition of the release of both beta glucuronidase and lysozyme from rat neutrophils and the release of both beta glucuronidase and histamine from mast cells were less than 1 microgram/ml. PMID- 7480176 TI - Biological activities of pyrrolidinoindoline alkaloids from Calycodendron milnei. AB - Certain genera of the tribe Psychotrieae, specifically Calycodendron and Psychotria, found on Pacific Islands, synthesize a series of Nb-methyltryptamine derived alkaloids made by linking together 2 to 8 pyrrolidinoindoline units. Nine alkaloids of this class have been isolated from the aerial parts and stem bark of Calycodendron milnei, a species endemic to the Vate Islands (New Hebrides), and examined for potential application as anti-cancer and anti-infective agents. All members of the series exhibited readily detected cytotoxic activity against proliferating and non-proliferating Vero African green monkey kidney cells in culture, with the most potent activity being exhibited by vatamine and quadrigemine C. Only hodgkinsine A exhibited substantial antiviral activity against a DNA virus, herpes simplex type 1, and an RNA virus, vesicular stomatitis virus. All members of the series showed readily detected anti bacterial, anti-fungal, and anti-candidal activities using both tube dilution and disc diffusion assay methods. The most potent anti-microbial alkaloids were hodgkinsine A and quadrigemine C, which exhibited minimum inhibitory concentration (MIC) values as low as 5 micrograms/ml. PMID- 7480177 TI - Physiological potential of beta-carotene in prolonging the survival of the host bearing transplantable murine lymphoma. AB - beta-Carotene, when supplemented in diet, has been found to increase the survival period of mice bearing a transplantable tumor, Dalton's lymphoma. Tumor cell count, body weight pattern, hematological parameters like total count showed marked alterations in a dose-responsive manner with beta-carotene administration when compared to their untreated counterparts. Decreased tumor cell proliferation is also reflected by increased hemoglobin levels of the host. PMID- 7480178 TI - Biological activities of cyanobacteria: evaluation of extracts and pure compounds. AB - A total of 80 lipophilic and hydrophilic extracts obtained from 20 samples of cultured freshwater and terrestrial cyanobacteria were investigated for their biological activities. Of all the extracts, 26% exhibited a significant lethal effect against brine shrimp. Out of 54 extracts tested for antimicrobial activity, 78% showed antibacterial and 45% antifungal activities. Of 30 extracts tested for cytotoxicity against KB cells, none was found to be active. Bioassay guided fractionation of the lipophilic extracts of Fischerella ambigua led to the isolation of three compounds; ambigols A (1) and B (2), and tjipanazole D (3). Compounds 1 and 2 exhibited antibacterial, antifungal, cytotoxic, molluscicidal, and anti-inflammatory, and antiviral activities. Tjipanazole D (3) showed moderate antibacterial properties. PMID- 7480179 TI - Antinociceptive properties of steroids isolated from Phyllanthus corcovadensis in mice. AB - The antinociceptive actions of the steroid compounds isolated from the leaves, stems, and roots of P. corcovadensis have been investigated in mice. Stigmasterol, stigmasterol acetate, beta-sitosterol, and aspirin (3-100 mk/kg, i.p.) inhibited, in a dose-related manner, acetic acid-induced abdominal constriction in mice with ID50s of 16, 11, 9, and 24 mg/kg, respectively. In the formalin test, stigmasterol and stigmasterol acetate (10-100 mg/kg, i.p.) caused graded inhibition of both the neurogenic (first phase) and inflammatory phases (second phase) of formalin-induced pain. However, both compounds were more effective in relation of the second phase of the formalin test with ID50 values of 26 and 41 mg/kg, respectively. Furthermore, both steroids failed to affect the edematogenic response of the formalin test. Given orally, stigmasterol and stigmasterol acetate (50-200 mg/kg) also exhibited significant though less potent analgesic action against both acetic acid- and formalin-induced nociception in mice. In addition, stigmasterol (up to 100 mg/kg, i.p.), in contrast to morphine (10 mg/kg, s.c.), had no analgesic effect in either tail-flick or hot-plate models. These findings suggest that stigmasterol and beta-sitosterol may account, at least in part, for the antinociceptive actions reported previously for the hydroalcoholic extract of Phyllanthus corcovadensis. PMID- 7480181 TI - Effect of peroxisomicine and related anthracenones on catalase activity. AB - Dimeric anthracenones were isolated from toxic plants of the genus Karwinskia (Rhamnaceae). T 514 or peroxisomicine A1 is one of these toxic compounds which produces an irreversible and selective damage on the peroxisomes of yeast cells in vivo. In this paper we now report the inhibitory effect in vitro of peroxisomicine A1 and other structurally related anthracenones on liver catalase activity. The peroxisomicine A1 produces a non-competitive inhibition with respect to H2O2 on bovine, dog, and mouse liver catalases. In the three cases Vmax was decreased whereas Km was unaffected. Other dimeric anthracenones of natural origin were also found to be inhibitors of bovine liver catalase. There is a relationship between structure and degree of inhibition of all anthracenonic compounds tested. Peroxisomicine A1 and peroxisomicine A2 caused the highest degree of inhibition (IC50 = 3.34 and 3.64 microM, respectively). PMID- 7480182 TI - Anti-diabetic effects of onion and garlic sulfoxide amino acids in rats. AB - On oral administration of onion (Allium cepa L.) and garlic (A. sativum L.) sulfoxide amino acids viz., S-methylcysteine sulfoxide (SMCS) and S-allylcysteine sulfoxide (SACS) to alloxan-diabetic rats for a month, their diabetic condition, being characterized by glucose intolerance, weight loss, depletion of liver glycogen, etc., was ameliorated as comparable to rats treated with glibenclamide and insulin. However, only the last two drugs significantly increased the conversion of labelled acetate to liver cholesterol which indicates a deterrent effect of the allium products against the synthesis of a risk factor. PMID- 7480183 TI - Characterization of the hypoglycemic effects of Trigonella foenum graecum seed. AB - The whole powder of Trigonella foenum graecum seeds and its extracts were tested for their hypoglycemic effect on normal and diabetic model rats. The powder, its methanol extract, and the residue remaining after methanol extraction had significant hypoglycemic effects when fed simultaneously with glucose. The water extract of the methanol extractive-free residue of the seed powder showed significant hypoglycemic activity at different prandial states. The Soluble Dietary Fibre (SDF) fraction showed no effect on the fasting blood glucose levels of nondiabetic or NIDDM model rats. However, when fed simultaneously with glucose, it showed a significant hypoglycemic effect (p < 0.05) in NIDDM model rats. Chemical analysis showed that the major constituent of the SDF is a galactomannan. The results confirm the involvement of SDF in the hypoglycemic effect of T. foenum graecum seeds. However, compound(s) other than SDF is (are) also involved in the hypoglycemic activity. PMID- 7480180 TI - Inhibition of lipid peroxidation and superoxide generation by diterpenoids from Rosmarinus officinalis. AB - Four diterpenoids, carnosic acid (1), carnosol (2), rosmanol (3), and epirosmanol (4), were isolated as antioxidative agents from the leaves of Rosmarinus officinalis by bioassay-directed fractionation. These diterpenoids inhibited superoxide anion production in the xanthine/xanthine oxidase system. Mitochondrial and microsomal lipid peroxidation induced by NADH or NADPH oxidation were also completely inhibited by these diterpenes at the concentration of 3-30 microM. Furthermore, carnosic acid protected red cells against oxidative hemolysis. These phenolic diterpenes were shown to be effective to protect biological systems against oxidative stresses. PMID- 7480184 TI - Antifungal principles of Baccharis pedunculata. AB - Four compounds including a flavone, an acetylenic lactone, a prenylated coumarin, and a 3-methyl ether flavone were isolated from the dichloromethane leaf extract of Baccharis pedunculata (Mill.) Cabr. (Asteraceae). The latter three compounds were identified to be responsible for the antifungal activity against some human pathogenic and phytopathogenic fungi. The most active compound, lachnophyllum lactone, an acetylenic lactone, showed a very high toxicity (LD50 2 micrograms/ml) against human keratinocytes. PMID- 7480185 TI - An antifungal naphthoquinone, xanthones and secoiridoids from Swertia calycina. AB - A chemical and biological screening of 25 species of the Gentianaceae family has been undertaken. Both methanolic and dichloromethane extracts of Swertia calycina exhibited a strong antifungal activity against Cladosporium cucumerinum and Candida albicans. The compound responsible for this activity has been isolated and identified as 2-methoxy-1,4-naphthoquinone. It is the first naphthoquinone to be described in Gentianaceae species. LC-UV and LC-TSP-MS analysis of the crude extracts of Swertia calycina also allowed on-line identification of six known xanthones and secoiridoids. PMID- 7480186 TI - Antimicrobial activity of tannins from Terminalia citrina. AB - Isolation of the fruit CH3OH extract of Terminalia citrina yielded five known tannins identified as corilagin (1) (3), punicalagin (2) (4), 1,3,6-tri-O-galloyl beta-D-glucopyranose (3) (5), chebulagic acid (4) (6), and 1,2,3,4,6-penta-O galloyl-beta-D-glucopyranose (5) (7) by comparison of their physical and spectral data with those of authentic samples. These tannins were tested for antimicrobial action. PMID- 7480187 TI - Studies on the inhibitory effects of some acridone alkaloids on Epstein-Barr virus activation. AB - Twenty-five acridone alkaloids from Citrus plants were examined for their inhibitory effects on Epstein-Barr virus activation by a short-term in vitro assay. 5-Hydroxynoracronycine (20) and acrimarine-F (25) showed remarkable inhibitory effects. PMID- 7480188 TI - Cytotoxicity of diterpenes from Premna schimperi and Premna oligotricha. AB - The cytotoxicity of two diterpenes from Premna schimperi and Premna oligotricha (Verbenaceae) was studied using the MTT assay. Their cytotoxic activity against three human (HeLa, SK.N.SH, and ECV 304) and two murine (L929 and RAW 264.7) carcinoma cell lines varied between 1.5 to 35 micrograms/ml and was comparable with azauridine and chlorambucil. PMID- 7480189 TI - Spasmolytic flavonols from Artemisia abrotanum. AB - Four flavonols with spasmolytic activity have been isolated from a methanol extract of Artemisia abrotanum L. (Asteraceae), as the principles primarily responsible for the smooth muscle relaxing activity of this plant. The flavonols show a dose-dependent relaxing effect on the carbacholine-induced contraction of guinea-pig trachea, the EC50 values for compounds 1-3 are 20-30 mumol/l while compound 4 is less active. PMID- 7480190 TI - Lipoxygenase inhibition and antioxidant properties of protoberberine and aporphine alkaloids isolated from Mahonia aquifolium. AB - Products of lipoxygenase metabolism play a role in the pathogenesis of psoriasis. Four protoberberine alkaloids, berberine, oxyberberine, jatrorrhizine, columbamine, and two aporphine alkaloids, magnoflorine, and corytuberine, isolated from Mahonia aquifolium, were tested for lipoxygenase inhibition. Oxyberberine, corytuberine, and columbamine were the most potent lipoxygenase inhibitors tested, whereas berberine and magnoflorine exhibited only low potencies. A strong linear correlation (r = 0.866) between lipoxygenase inhibition and lipid antioxidant properties of these compounds was found. These data suggest that the mechanism of lipoxygenase inhibition by these alkaloids may be linked to the inhibition of lipid hydroperoxide substrate accumulation. Inhibition of lipoxygenase by these compounds may contribute to the therapeutic effect of M. aquifolium extracts in the treatment of psoriasis. PMID- 7480191 TI - Sesquiterpene lactones and other constituents of Centaurea napifolia. AB - Aerial parts of Centaurea napifolia furnished the sesquiterpene lactones cnicin, 4'-acetoxycnicin, melitensin, dehydromelitensin, two known esters of dehydromelitensin, lappaol and a lappaol isomer, and a 1,2-diacylated glucose. PMID- 7480192 TI - Rearranged taxoids from Taxus yunnanensis. AB - Two rearranged 2(3-->20)abeotaxanes were isolated from the leaves and stems of Taxus yunnanensis, and designated as taxin B and 2-deacetyltaxin B. The structure of taxin B with a unique 6/10/6 skeleton was deduced on the basis of 1H-NMR, 13C NMR, 1H-1H COSY, DEPT, NOE, and mass spectra and confirmed by X-ray crystallographic analysis. The structure of 2-deacetyltaxin B is elucidated by analog. PMID- 7480193 TI - Isorhamnetin sulphate from the leaves and stems of Oenanthe javanica in Korea. PMID- 7480194 TI - Isolation and antibacterial activity of marchantin A, a cyclic bis(bibenzyl) constituent of Hungarian Marchantia polymorpha. PMID- 7480195 TI - Ineupatorolides from Carpesium glossophyllum. PMID- 7480196 TI - Mayteine and 6-benzoyl-6-deacetylmayteine from Maytenus krukovii. PMID- 7480197 TI - Effect of schisandrin B on hepatic glutathione antioxidant system in mice: protection against carbon tetrachloride toxicity. AB - Pretreating female Balb/c mice with schisandrin B (Sch B) at increasing daily doses (1-4 mmol/kg) for 3 days caused dose-dependent increases in hepatic glutathione S-transferase (GST) and glutathione reductase (GRD) activities. However, the activities of glucose-6-phosphate dehydrogenase (G6PDH), Se glutathione peroxidase (GPX), and gamma-glutamylcysteine synthetase (GCS) were down-regulated to varying degrees in a dose-dependent manner. While there were biphasic changes in hepatic reduced glutathione (GSH) level as well as susceptibility of hepatic tissue homogenates to in vitro peroxide-induced GSH depletion, a gradual decrease in hepatic malondialdehyde content was observed. The beneficial effect of Sch B on the hepatic GSH anti-oxidant system became more evident after CCl4 challenge. The same Sch B pretreatment regimen caused a dose dependent protection against carbon tetrachloride (CCl4)-induced hepatotoxicity. The hepatoprotection was associated with significant enhancement in hepatic GSH status, as indicated by the substantial increase in tissue GSH levels and the corresponding decrease in susceptibility of tissue homogenates to GSH depletion. Where the activities of GST and GRD were increased linearly over non-CCl4 control values, there was also a gradual elevation in G6PDH activity upon administration of increasing doses of Sch B. In contrast, GPX activity was moderately down regulated. The ensemble of results suggests that the hepatoprotection afforded by Sch B pretreatment may mainly be attributed to the enhancement in the functioning of the hepatic GSH anti-oxidant system, possibly through stimulating the activities of GSH related enzymes. PMID- 7480198 TI - Bellidifolin stimulates glucose uptake in rat 1 fibroblasts and ameliorates hyperglycemia in streptozotocin (STZ)-induced diabetic rats. AB - Bellidifolin, isolated from Swertia japonica, was found to be a potent hypoglycemic agent in STZ-induced diabetic rats by both oral and intraperitoneal administration. Bellidifolin significantly lowered the loaded glucose level in normal as well as diabetic rats. Bellidifolin also lowered blood triglyceride levels significantly. It stimulated glucose uptake activity in Rat 1 fibroblasts expressing human insulin receptors. PMID- 7480199 TI - Emodin pharmacokinetics in rabbits. AB - Administration of emodin to rabbits by i.v. bolus resulted in a serum profile which could be well described by a two-compartment model. The AUC of emodin was 518 micrograms.min/ml, clearance was 72.3 ml/min, and elimination half life was 227 min which was much longer than that reported in a previous study. Oral administration of emodin resulted in a very low serum concentration. Protein binding of emodin was investigated by the equilibrium dialysis method. Emodin was found to be highly bound (99.6%) to serum protein. PMID- 7480201 TI - Inhibitors of DNA topoisomerase I and II isolated from the Coptis rhizomes. AB - A water extract of Coptis chinensis was found to have the ability to stabilize the cleavable complex with mammalian DNA topoisomerase I. As the result of bioassay-guided fractionation, two protoberberine alkaloids, epiberberine and groenlandicine, were identified as active principles with topoisomerase I mediated DNA cleavage activity in vitro. These two alkaloids did not induce topoisomerase II-mediated DNA cleavage. During further examination of the structurally related protoberberine alkaloids, berberrubine which is produced during the processing of Coptis rhizome as traditional medicine, was identified as a specific inducer of topoisomerase II-mediated DNA cleavage in vitro. These results indicated that protoberberine alkaloids are a chemical family which can induce cleavable complexes with topoisomerases I and II. PMID- 7480200 TI - Reversal of daunomycin and vinblastine resistance in multidrug-resistant P388 leukemia in vitro through enhanced cytotoxicity by triterpenoids. AB - Examined in vitro were the effects of some triterpenoids from Panax (Araliaceae) and Glycyrrhiza (Leguminosae) spp. on the sensitivity to daunomycin (DAU) and vinblastine (VBL) of adriamycin (ADM)-resistant P388 leukemia cells (P388/ADM), which were resistant to multiple anticancer drugs. Quasipanaxatriol, 20(S) protopanaxatriol, ginsenoside Rh2, and compound K greatly enhanced the cytotoxicity of the anti-cancer drugs in P388/ADM cells. The extent of enhancement was different among the triterpene compounds; the 4- to 46-fold increase in DAU cytotoxicity was observed in P388/ADM cells in the presence of non-toxic or marginally toxic concentrations of individual compounds, while those for VBL were in the ratios of 2- to 37-fold. The maximum increase in cytotoxicity was observed with 50 microM quasipanaxatriol; the resistance indices defined to be the ratios of the IC50 values for P388/ADM and P388 parental cells decreased from 79 to 1.7 and from 180 to 4.9 in the cases of DAU and VBL, respectively. The reversal of DAU resistance in P388/ADM by quasipanaxatriol could be explained by the effective accumulation of the drugs mediated by the DAU-efflux blockage. PMID- 7480202 TI - Anti-herpes virus activity of aporphine alkaloids. AB - We evaluated, in cell cultures, the action of a series of 19 aporphine alkaloids against Herpes simplex virus type 1 (HSV-1). On the basis of viral titre reduction, six alkaloids were found to be active. The mode of action of the three most potent inhibitors, oliverine HCl, pachystaudine, and oxostephanine, was studied. These compounds did not have any virucidal or prophylactic effect but they were shown to interfere with the viral replicative cycle. Although DNA synthesis was reduced, their exact target remains to be elucidated. In the discussion, some structure-activity relationships are considered. PMID- 7480203 TI - Potent inhibitors of tyrosinase activity and melanin biosynthesis from Rheum officinale. AB - Thirty-three crude drug extracts were screened for their tyrosinase inhibitory activity. Among them, the acetone extract of the rhizomes of Rheum officinale Baillon showed the strongest inhibitory activity. Tyrosinase inhibitory activity guided fractionation and chemical analysis led to the isolation of two potent compounds, 3,4',5-trihydroxystilbene-4'-O-beta-D-(2"-O-galloyl)glucopyr anoside (1) and 3,4',5-trihydroxystilbene-4'-O-beta-D-(6"-O-galloyl)glucopyr anoside (2). These compounds showed a competitive inhibition against tyrosinase and also inhibited the melanin biosynthesis. PMID- 7480204 TI - Effect of oral administration of a pectic polysaccharide fraction from a kampo (Japanese herbal) medicine "juzen-taiho-to" on antibody response of mice. AB - A dried decoction of a kampo (Japanese herbal) medicine, Juzen-Taiho-To (TJ-48), has been fractionated into five fractions and tested for their effects on antibody response of mice. An intraperitoneal injection (300 mg/kg) of TJ-48 stimulated anti-sheep red blood cell (SRBC) antibody response of normal Balb/c mice, but only the polysaccharide fraction (F-5, 300 mg/kg) enhanced the antibody response among the fractions from TJ-48. When F-5 (0.5 or 1.0 g/kg/day) was orally administered to normal Balb/c mice (7-weeks-old) from 7 days before to 4 days after immunization with SRBC, the number of anti-SRBC-IgM-PFC in spleen and the titer of anti-SRBC-IgM in plasma were increased significantly. However, a lower dose (0.1 g/kg/day) of F-5 did not show a significant stimulative activity on the anti-SRBC-response. Although aged Balb/c mice (6-months-old) produced a lower level of anti-SRBC-IgG in comparison with young Balb/c mice (8-weeks-old), the anti-SRBC-IgG response of the aged mice was stimulated significantly when F-5 (0.13 g/kg/day) or TJ-48 (1.0 g/kg/day) was orally administered to the aged mice from 6 days before immunization. Intraperitoneal injections of i-carrageenan (2.5 mg/kg/day) at 3 and 1 days before the immunization with SRBC increased the level of anti-SRBC antibody response compared with normal mice. Oral administrations of TJ-48 (1.0 g/kg/day) or F-5 (0.5 g/kg/day) to the i-carageenan-treated mice reduced the level of the anti-SRBC-antibody response near to that of normal mice.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7480205 TI - Radical scavenger activity of different 3',4'-dihydroxyflavonols and 1,5 dicaffeoylquinic acid studied by inhibition of chemiluminescence. AB - To gain more insights into structure-activity relationships, four 3',4' dihydroxyflavonols differing in the substitution of the A and C rings and 1,5 dicaffeolyquinic acid were evaluated for their ability to inhibit chemiluminescence of human neutrophils stimulated with opsonized zymosan or FMLP as well as in an enzymatic system with H2O2 and horseradish peroxidase. It could be shown that an additional o-dihydroxy structure in the A-ring, or a 6-methoxy group, respectively, has no significant influence, thus confirming the o dihydroxy group of the B-ring as the most important structural feature for the radical scavenger activity. It can be supposed that the main effect of the tested flavonols is based on their inhibition of myeloperoxidase, besides inhibition of enzymes involved in activating the NADPH-oxidase, and a direct reaction with oxygen radicals. Inhibition of chemiluminescence by 1,5-dicaffeoylquinic acid was in the same order as those observed with the flavonols. PMID- 7480207 TI - Triterpenoid glycosides from the fruits of Kochia scoparia. AB - From the fruits of Kochia scoparia (L.) Schrad, five triterpenoid glycosides were isolated for the first time from this plant. They were elucidated as momordin Ic, the 6'-methyl ester of momordin Ic, momordin IIc, 2'-O-beta-D glucopyranosylmomordin Ic, and 2'-O-beta-D-glucopyranosylmomordin IIc on the basis of spectral and chemical methods. The last two saponins are new natural products. PMID- 7480206 TI - Asteryunnanosides F and G: two new triterpenoid saponins from Aster yunnanensis. AB - Two new triterpenoid saponins, aster-yunnanosides F and G, were isolated from the roots of Aster-yunnanensis. Their structures were determined as oleanolic acid-28 O-beta-D-glucopyranosyl-(1-->6)-beta-D-glucopyranoside and 3-O-beta glucopyranosyloleanolic acid-28-O-beta-D-glucopyranosyl-(1-->6)-[alpha-L rhamnopyranosyl-(1-->2) ]- beta-D-glucopyranoside by spectral data, especially 2D NMR analysis including COSY, HETCOR, HOHAHA, and ROESY techniques, and chemical transformation. PMID- 7480208 TI - Determination of strychnine and brucine by capillary zone electrophoresis. AB - A procedure for quantitative estimation of strychnine and brucine in the extracts of Strychnos nux-vomica seeds by capillary zone electrophoresis (CZE) was developed. The buffer solution used was 10mM phosphate buffer-MeOH (9:1), pH 2.5. The linear calibration range was 0.01-0.15 mg/ml. This method is useful for the qualitative and quantitative determination of strychnine and brucine in plant drug samples, as well as in human plasma. PMID- 7480209 TI - Authentication of Panax species and their adulterants by random-primed polymerase chain reaction. AB - DNA from dried or fresh roots of three medicinal Panax species and their adulterants were amplified by AP-PCR or RAPD. Fingerprints for P. ginseng or P. quinquefolius were found to be consistent irrespective of plant source or age. On the other hand, very different fingerprints were obtained from adulterants. Hence the PCR approach may be used as a means to authenticate the concerned Panax species. Analysis of the degree of similarity of the fingerprints confirmed that P. ginseng is more closely related to P. quinquefolius than to P. notoginseng. PMID- 7480210 TI - A cytotoxic diacetylene from Dendropanax arboreus. AB - The crude ethanol extract from the leaves of Dendropanax arboreus (Araliaceae) from Monteverde, Costa Rica, exhibits cytotoxic activity against Hep-G2, A-431, H 4IIE, and L-1210 tumor cell lines, but is not toxic against normal hepatocytes. The active component has been isolated by activity-directed separation and identified by 1H- and 13C-NMR spectroscopy as the acetylenic compound cis-1,9,16 heptadecatriene-4,6-diyne-3,8-diol. PMID- 7480211 TI - Effects of catechins on the mouse tumor cell adhesion to fibronectin. AB - We studied the effects of 5 kinds of catechins on the adhesion of mouse lung carcinoma 3LL and melanoma B16F10 cells to the fibronectin substratum. (-) Epicatechin gallate and (-)-epigallocatechin gallate were active in inhibiting the 3LL cell adhesion, while (+)-catechin, (-)-epicatechin, and (-) epigallocatechin were inactive. Gallate-containing catechins also impaired adhesion and/or spreading of B16F10 cells. PMID- 7480212 TI - Screening for in vitro antifungal activities of some indole alkaloids. AB - Several indole alkaloids were tested by the agar dilution technique on a panel of human pathogenic fungi, yeasts, and dermatophytes. Our results indicate that the most active compounds possess a beta-carboline skeleton and that the presence of a 3-4 double bond enhances the activity. Our results also show that antifungal activities are not linked to the cytotoxic, antimicrobial or antiparasitic properties. PMID- 7480213 TI - Effects of honokiol and magnolol on acetylcholine release from rat hippocampal slices. AB - To study the possible mechanism through which honokiol and magnolol elicit their central depressant effects, we examined the influence of these two phenolic compounds on 25 mM K(+)-stimulated release of [3H]acetylcholine (ACh) from the rat's hippocampal slices. Honokiol, but not magnolol, elicited a concentration dependent enhancement of K+-evoked ACh release. Addition of either tetrodotoxin, pilocarpine, or methoctramine had no effect on honokiol-enhanced ACh release. These results suggest that honokiol enhanced K(+)-evoked ACh release directly on hippocampal cholinergic terminals via receptors other than the M2 cholinergic subtypes. PMID- 7480215 TI - Bistetrahydrofuranoid acetogenins from Rollinia sericea. PMID- 7480216 TI - Lignans from barks of Hernandia nymphaefolia and H. peltata. PMID- 7480214 TI - Phenylethanoids in the herb of Plantago lanceolata and inhibitory effect on arachidonic acid-induced mouse ear edema. AB - The five phenylethanoids, acteoside (1), cistanoside F (2), lavandulifolioside (3), plantamajoside (4) and isoacteoside (5) were isolated from the herb of Plantago lanceolata L. (Plantaginaceae). Compounds 1, the major phenylethanoid in the herb of P. lanceolata L., and 4, the major phenylethanoid in the herb of P. asiatica L., showed inhibitory effects on arachidonic acid-induced mouse ear edema. PMID- 7480217 TI - Flavonoids of Artemisia austriaca. PMID- 7480218 TI - Luteolin 7-diglucuronide, the major flavonoid compound from Aloysia triphylla and Verbena officinalis. PMID- 7480219 TI - Tannin constituents of Terminalia glaucescens. PMID- 7480220 TI - Glucuronidation of magnolol assessed using HPLC/fluorescence. PMID- 7480221 TI - The fetal fibroblast: the effector cell of scarless fetal skin repair. AB - Human fetal skin heals without scar formation when it is transplanted to a subcutaneous location on an adult athymic mouse and subsequently wounded. In contrast, human fetal skin of identical gestational age heals with scar formation when transplanted to a cutaneous location on the athymic mouse recipient. To determine if mouse (adult) or human (fetal) fibroblasts are healing the graft wounds, we performed indirect immunohistochemistry for mouse and human collagen types I and III. Full-thickness skin grafts (n = 51) from human fetuses at 18 weeks' (n = 4) or 24 weeks' (n = 2) gestational age were placed onto athymic mice in two locations: cutaneously onto a fascial bed and subcutaneously in a pocket under the murine panniculus carnosus. Linear incisions were made in each graft 7 days after transplantation. Grafts were harvested at 7, 14, and 21 days after wounding and stained with hematoxylin and eosin or Mallory's trichrome. Immunohistochemistry for either human collagen type I or type III or for mouse collagen type I was performed. The subcutaneous grafts healed with human collagen types I and III in a scarless pattern. The wound collagen pattern was reticular and unrecognizable from the surrounding dermis. Hair follicles and sebaceous gland patterns were unchanged in the wounded dermis. Conversely, the cutaneous grafts healed with mouse collagen in a scar pattern with disorganized collagen fibers and no appendages. Mouse collagen scar was present along the base of the cutaneous grafts and as a thin capsule around the subcutaneous grafts. We conclude that (1) subcutaneous grafts heal with human fetal collagen and no scar formation, and (2) cutaneous grafts heal with mouse collagen in a scar pattern. Fetal fibroblasts can heal fetal skin wounds without scar despite being perfused by adult serum and inflammatory cells in an adult environment. These data suggest that the fetal fibroblast is the major effector cell for scarless fetal skin repair. PMID- 7480222 TI - The palatal island flap on retrospection. AB - The place of the palatal island flap in present-day surgery is evaluated. Attention is drawn to the hitherto unrecognized fact that the first palatal island flap was executed decades before its general introduction in 1962. Its origins are discussed, and the subsequent process from its use as a panacea to the gradual evolution of its indications is described. Variations in anatomy, complications, success rates, and contraindications are reviewed. Attention is paid to the problems inherent in studies of palate surgery, both retrospective and prospective. A balance is made up of what has and has not been proven adequately. To date, no advantages over other palate-lengthening procedures have been established, while there is some evidence that it might in fact be deleterious to maxillary growth. As a result, at present, the palatal island flap is used mainly in closure techniques, as in fact was done by the first two surgeons using that it more than half a century ago. PMID- 7480223 TI - Anatomic considerations in transconjunctival blepharoplasty. AB - To better identify the anatomy of the lower eyelid in relation to the transconjunctival approach to blepharoplasty, 20 fresh cadaver lower eyelids were dissected. Cross-sectional cuts of fresh cadaver specimens also were made and examined by whole-organ photomicrographs of the entire lower eyelid and its associated soft-tissue structures. These were correlated with magnetic resonance imaging studies in the living patient. Measurements were made of the relationship of the lid margin, tarsal plate, infraocular fat (including the three traditional fat compartments), inferior oblique muscle, and eyelid (on stretch). Our findings suggest that a transconjunctival incision closer to the fornix, directed toward the infraorbital rim, allows the surgeon a direct and safe approach to the fat compartments. No distinct anatomic compartmentalization of the periorbital fat as traditionally described was found. There was a consistent extension of the lateral pad, lateral to the lateral canthus of the eye. There are also accumulations of fat outside the orbital septum in the cheek and beneath the orbicularis muscle that contribute to the fullness of the lower eyelid and which cannot be addressed by the transconjunctival approach. PMID- 7480224 TI - Anterior SMAS plication for the treatment of prominent nasomandibular folds and restoration of normal cheek contour. AB - Prominent nasomandibular folds and cheek concavities are common sequelae of the aging process. The difficulty in achieving initial and long-lasting correction has been well documented. Anterior SMAS plication as an adjunct to subcutaneous rhytidectomy has proven to be an effective, straightforward method of flattening and rejuvinating the nasomandibular folds. Ptotic cheek fat is returned to the cheek hollow, and the end result is a more youthful, symmetrical flattening of the nasomandibular folds and jowls. The anterior SMAS is plicated vertically with a row of permanent sutures placed just lateral to the nasomandibular fold. Less tension is required on the skin flaps at closure, and this permits precise control of anterior SMAS vectors without the time, expense, and potential morbidity of more extensive dissections. The additional cost is that of two sutures and an extra 10 to 15 minutes of operative time. This technique has been performed on 226 patients of both sexes, and there have been no complications, including nerve injuries, infections, or dehiscences. The longest follow-up has been 2.2 years, and there have been no asymmetries, contour irregularities, or early recurrences. PMID- 7480225 TI - Unusual complications of the Le Fort I osteotomy. AB - In this report we describe three very different and interesting complications of the Le Fort I osteotomy. The first case is of a unilateral third nerve palsy developing as a result of a carotid-cavernous fistula and false aneurysm of the internal carotid artery. The second complication to be reported is total unilateral blindness. The third case is one of total avulsion of a lateral segment of the palate in a patient with a complete bilateral cleft of the primary and secondary palates. All complications were the result of a Le Fort I osteotomy performed for correction of midface hypoplasia secondary to congenital cleft lip and palate. The literature is reviewed, and the pathomechanics of each injury are hypothesized. PMID- 7480226 TI - Consideration of operative procedures for zygomatic reduction in Orientals: based on a consecutive series of 28 clinical cases. AB - Operative procedures for reduction malar bone plasty were considered in our consecutive series of 28 Japanese cases. The patients consisted of 7 males and 21 females whose ages ranged from 21 to 57 years. The operative procedures used were either shaving the protruded zygomatic body and arch with chisels by means of an intraoral approach and/or an extraoral sideburn-lateral canthal approach or, alternatively, mobilizing the protruded zygoma en bloc medioposteriorly into reduction of the prominent zygoma by a coronal approach. In principle, the noncoronal approach was indicated for males and young females, and the coronal approach, conducted in association with forehead lifting with or without canthopexy, was indicated for females over age 40. These procedures have individual merits and demerits. In some cases, associated surgery was performed concomitantly according to the facial balance or facial rejuvenization. We believe that the optimal operative approach and operative procedure should be considered in accordance with the patient's sex, age, wishes, and/or cultural background. PMID- 7480227 TI - Morbidity and functional outcome of free jejunal transfer reconstruction for circumferential defects of the pharynx and cervical esophagus. AB - Free jejunal transfer has been criticized by some surgeons as unreliable, poorly tolerant of radiation therapy, and associated with significant morbidity and dysphagia. To determine the validity of these criticisms, we reviewed 93 patients who underwent 96 free jejunal transfers for repair of circumferential pharyngoesophageal defects over a 5-year period. The free jejunal transfer success rate was 97 percent; all 3 failures were repaired with repeated free jejunal transfer. The complication rate was 57 percent (55 of 96); fistula (19 percent) and stricture (15 percent) were the most common complications. An oral diet was tolerated by 80 percent of patients; 85 percent of these resumed their diet within 2 weeks of surgery. Causes of dysphagia were multifactorial. Postoperative radiation therapy did not increase morbidity or dysphagia. The perioperative mortality rate was 2 percent. We conclude that free jejunal transfer is a reliable method of pharyngoesophageal reconstruction that has an acceptable morbidity rate and a low mortality rate. Postoperative radiation therapy is well tolerated by the free jejunal transfer. PMID- 7480228 TI - The effect of Biocell texturing and povidone-iodine irrigation on capsular contracture around saline-inflatable breast implants. AB - We performed a prospective, concurrently controlled, and blinded 4-year clinical study on 60 patient volunteers to determine the effects of two independent variables, McGhan's Biocell texturization and Betadine antibacterial irrigation, on the incidence of fibrous capsular contracture around saline-inflatable implants following retromammary augmentation. Each patient was randomly assigned both a textured and a smooth implant and both saline and Betadine irrigation so that each patient served as her own control. The textured devices irrigated with Betadine experienced an overall incidence of contracture of only 4 percent compared with 50 percent for the smooth devices irrigated with saline solution. The Betadine-irrigated devices in general had a lower incidence of contracture than the saline-irrigated devices, and the textured-surface devices in general had a lower incidence of contracture than the smooth devices. Antibacterial irrigation and surface texturization may work in a cumulative manner to reduce the early incidence of capsular contracture. PMID- 7480229 TI - Biodegradation of polyurethane foam, revisited, in the rat model. AB - In a prospective, randomized, controlled animal study, we systematically analyzed implanted polyurethane foam to determine the rate of degradation in the rat and to compare it to our previous human data. Sixteen 1-cm2 silicone-backed pieces of polyurethane foam were randomly implanted into dorsal subcutaneous pockets in each of 16 Sprague-Dawley rats. Eight animals had polyurethane implants removed at 3 and 6 months and the remaining 8 animals at 9 and 12 months. Specimens were examined either by histology or by scanning electron microscopy of recovered foam after collagenase digestion of the capsule. Histologically, there was evidence of foam degradation and a multinucleated giant cell inflammatory response surrounding the implants. With the scanning electron microscope, the strut width of the polyurethane foam upon implantation measured 51.4 +/- 1.3 microns (mean+SEM). This progressively decreased to 29.1 +/- 1.3 microns at 3 months, 16.6 +/- 0.6 micron at 6 months, 14.9 +/- 0.5 micron at 9 months, and 13.2 +/- 0.3 micron at 12 months (p < 0.0001). Duration of implantation has a significant impact on polyurethane degradation as measured by scanning electron microscopy in the rat animal model. The rate of degradation in the rat is much faster than in our human study, indicating that the rat studies cannot be used to draw conclusions regarding rate of biodegradation in humans. PMID- 7480230 TI - An anatomic study for the rectus abdominis myocutaneous flap combined with a vascularized rib. AB - The anatomy of the vascular connections between the eighth intercostal vessels and the deep epigastric vessels was studied in a series of 20 dissections in 10 preserved cadavers to develop the rectus abdominis myocutaneous flap combined with a vascularized rib. The distance between the point where the eighth intercostal vessels attached to the costal margin and the xiphoid process and the arcuate line, which become a good guide for design of the flap, was measured. The average length of the vascular pedicle of a vascularized rib was over 10 cm. Possible clinical uses of this compound flap for reconstruction of a composite maxillary or mandibular defect are described. PMID- 7480231 TI - Extended skin island delay of the unipedicle TRAM flap: experience in 35 patients. AB - A technique to surgically delay a single-pedicle TRAM flap is described. This extended skin island delay essentially divides the unipedicle TRAM flap into two stages separated by 1 week. Experience with this flap in 35 patients is reported. On the basis of this experience, we believe that the extended skin island delayed TRAM flap should be considered a safe and reliable alternative to double muscle pedicle and free tissue transfer in high-risk patients. PMID- 7480232 TI - Hernia prevention and aesthetic contouring of the abdomen following TRAM flap breast reconstruction by the use of polypropylene mesh. AB - The value of synthetic mesh use in the treatment of recurrent abdominal hernias is well recognized and has led to its advocacy by some authors as an adjunct in primary hernia repair. Mesh use in the donor-site closure associated with TRAM flap reconstruction is typically restricted to situations where undue tension or questionable tissue integrity may be predisposing factors to herniation. Although more liberal use of mesh has been advocated for these circumstances, fear of mesh complications may continue to restrict its use. We present a series of 65 consecutive patients who had routine mesh application to fascial closures following TRAM flap breast reconstruction. The use of mesh provides an added margin of strength to fascial reconstruction and was found to have additional benefit as a technical adjunct to the aesthetic aspects of the abdominoplasty. Mean patient follow-up was 56.4 months. The resulting rates of hernia (1.5 percent) and mesh-related infection (1.5 percent) demonstrate its considerable safety. We recommend consideration of polypropylene mesh use for improved strength and aesthetic quality of the donor-site closure following TRAM flap breast reconstruction. PMID- 7480233 TI - Evidence for the efficacy of TRAM flap delay in a rat model. AB - The dilemma in selecting a variant of the transverse rectus abdominis musculocutaneous (TRAM) flap for autogenous tissue breast reconstruction is to balance maximal flap perfusion against the sacrifice of abdominal-wall integrity. A surgical delay has been suggested as one very simple fascia-sparing technique that simultaneously augments flap volume, but no randomized clinical studies have been performed to prove whether there is any benefit whatsoever following such a maneuver. Because large groups of nearly identical subjects may be more readily available for comparison in an animal model, the rat TRAM flap provides an inexpensive method to further investigate the delay phenomenon in this setting. Staged procedures were performed in 15 Sprague-Dawley rats initially causing a delay by division of one or both dominant vessels (cranial epigastric) supplying the rectus abdominis muscles. Either 2 or 4 weeks later, TRAM flaps were elevated based only on the subservient (caudal epigastric) pedicle. A control group of 7 rats with immediate formation of a TRAM flap had a mean survival of 46.3 +/- 15.37 percent of their original flap surface area. After a 2-week delay, viability was 50.2 +/- 17.54 percent for ipsilateral and 39.4 +/- 7.57 percent for flaps with prior bilateral dominant pedicle division. Neither was significantly different from control (p = 0.68 and 0.38, respectively). However, the ipsilateral 4-week delayed group had 67.8 +/- 8.96 percent flap survival, which represented a significant enhancement when compared with the undelayed control TRAM flaps (p < 0.02).(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7480234 TI - Preoperative and postoperative evaluation of latissimus dorsi myocutaneous flap vascularization by color flow duplex scanning. AB - At the Plastic and Reconstructive Surgery Department of the University of Perugia, 104 patients have been examined by color flow duplex scanning in order to study latissimus dorsi vascular supply before and after surgical procedures. This noninvasive ultrasound technique provides a precise and accurate measurement of blood flow velocity (15 cm/s) and volume (27 ml/min) within the thoracodorsal artery, which represents the vascular pedicle of the flap, and it allows us to detect the exact location of the perforating branches with their cutaneous distribution. This preoperative evaluation, pointing out vascular pathologies that contraindicate use of the latissimus dorsi flap, can be considered a great help for the reconstructive surgeon in the choice of the most suitable operation, and it can surely lead to a significant decrease in postoperative graft failure rates. PMID- 7480235 TI - The gluteus maximus muscle-splitting myocutaneous flap for treatment of sacral and coccygeal pressure ulcers. AB - We describe a modified technique using the gluteus maximum muscle as a splitting myocutaneous flap to close specifically low sacral and coccygeal pressure ulcers. Twenty-eight patients with sacral or coccygeal stage IV pressure ulcers (average size 4 x 4 cm) underwent a gluteus maximus muscle-splitting myocutaneous flap when conservative treatment failed to heal the ulcer. Twenty-seven of the 28 patients had complete healing of the pressure ulcer site at an average follow-up of 15 months (range 2 to 40 months). Complications occurred in 7 patients, requiring revision of the flap in 2 patients. The advantages of this technique include reduced blood loss, preservation of most of the gluteus maximus for future use, and retained function of the gluteus maximus for stair climbing and single-limb support in the ambulatory patient. We recommend the gluteus maximus muscle-splitting myocutaneous flap as the procedure of choice for closure of small low sacral or coccygeal ulcers in both the ambulatory and nonambulatory patient. PMID- 7480236 TI - The fallacy of presumed superiority of proximally based versus distally based flaps. AB - As our knowledge of the origin of the cutaneous circulation becomes more precise, many preconceived notions continue to be abandoned. In this context, the general rule that distally based flaps are inherently inferior to proximally based flaps also should appropriately be challenged. To disprove this long-standing dictum, multiple abdominal flaps were simultaneously elevated in 15 Sprague-Dawley rats that included or excluded all possible combinations of musculocutaneous or axial skin perforators from each major source vessel (cranial or caudal epigastrics, superficial epigastric or lateral thoracic, respectively). These flaps were sequenced randomly in each rat to have superior (proximal) or inferior (distal) pedicles such that at least five flaps of each subtype were analyzed. Mean flap survival when the specific source vessel was retained uniformly was significantly greater than in all flaps where the source vessel had been excluded regardless of the orientation of the flap pedicle. All flaps with intact circulation had no significant difference in mean flap survival. Similarly, when normal flap circulation was excluded, no difference was observed in mean flap survival. Again, both these observations were true irrespective of the location of the flap base. Thus the major factor in determining flap viability was not whether its pedicle was proximally or distally based but rather the origin and quality of its intrinsic blood supply. PMID- 7480237 TI - A prospective, double-blind, placebo-controlled trial of a single dose of azithromycin on postoperative wound infections in plastic surgery. AB - Over a 9-month period from September of 1991 to May of 1992, 339 patients were included in a randomized, double-blind, placebo-controlled study using azithromycin as the prophylactic agent to determine whether it effects a clinically meaningful reduction in postoperative surgical infections in plastic surgery. Azithromycin was given as prophylaxis in 171 patients and placebo in 168 patients. The study medication was a single oral dose taken at 8 P.M. the day before surgery. The patients were followed up for a minimum of 4 weeks after surgery. The patients who received wound infection prophylaxis had 5.1 percent infections compared with 20.5 percent in the placebo group (p = 0.00009). Eighty percent of all wound infections were first seen after discharge, explaining why plastic surgeons might overlook their infectious complications. There was a significant reduction in postoperative complications (p = 0.04) and in the additional use of antibiotics postoperatively (p = 0.007) in the prophylaxis group. Subgroup analysis showed a significant reduction in surgical infections in breast surgery (p < 0.05) and reconstructive surgery with flaps (p < 0.05). No effect of the prophylactic regime was demonstrated in patients undergoing secondary surgery for cleft lip and palate disease. PMID- 7480238 TI - Multiple primary melanomas: data and significance. AB - Cancer statistic reports show that the incidence of melanoma has increased each decade. It is now estimated that approximately 5 percent of the patients with primary cutaneous melanoma will develop another primary melanoma in their lifetime. This report describes the information gathered from 27 patients at the Yale Melanoma Unit who have developed 59 individual primary melanomas; 22 of the patients developed a second primary melanoma, and 5 patients each developed three primary melanomas. In 8 patients (30 percent), the second primary melanoma was diagnosed within 1 month of the first malignancy and was considered synchronous. The remaining 24 melanomas in the 19 patients presented subsequently: 4 (17 percent) within the first year, 7 (29 percent) during the second year, and 13 (54 percent) beyond the second year of the first diagnosis. Although the thickness of the initial melanoma ranged from 0.2 to 6.0 mm, all subsequent melanomas were either in situ or less than 1.0 mm in thickness. This study shows that patients who developed more than one melanoma invariably had thin subsequent lesions. The implications of the multiple melanomas are not a poorer prognosis, but rather that the patients' prognosis is the same as that of the original, or thickest, melanoma. PMID- 7480239 TI - Injectable cartilage. AB - Slowly polymerizing calcium alginate gels were investigated as a means of delivering large numbers of isolated chondrocytes by means of injection to determine if these gels would promote engraftment and could provide three dimensional templates for new cartilage growth. Chondrocytes isolated from the articular surface of calf forelimbs were added to a 1% sodium alginate dissolved in a 0.1 M potassium phosphate buffer solution (pH 7.4) to generate a final cellular density of 10 x 10(6)/ml (representing approximately 10 percent of the cellular density of human juvenile articular cartilage). The calcium alginate chondrocyte mixture was injected through a 22-gauge needle in 100-microliters aliquots under the panniculus carnosus on the dorsum of nude mice and incubated for 6 (n = 4), 8 (n = 11), and 12 (n = 12) weeks in vivo. Time-zero specimens (n = 10) consisting of 100-microliters aliquots of the calcium alginate-chondrocyte mixture were used to calculate initial weight. At harvest, all calcium alginate chondrocyte specimens exhibited a pearly opalescence and were firm to palpation as early as 6 weeks after injection. By 12 weeks of in vivo incubation, the specimens weighed 0.15 +/- 0.04 gm, significantly more than the initial weight of 0.11 +/- 0.01 gm (p < 0.05). Specimens stained with hematoxylin and eosin demonstrated lacunae within a basophilic ground-glass substance. Control specimens of calcium alginate without chondrocytes (n = 4) had a doughy consistency 12 weeks after injection and had no histologic evidence of cartilage formation.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7480240 TI - Telescoping and glue technique in vein grafts for arterial defects. AB - Fibrin glue was applied at the anastomotic site of the vein graft that was placed in rat femoral artery defects by the telescoping anastomotic technique at both proximal and distal anastomoses. All 35 vein grafts thus placed were patent, yielding a statistically higher patency rate than those placed in the same femoral artery by the same telescoping anastomotic technique but without fibrin glue. The telescoping anastomotic technique assisted by fibrin glue appears to give as reliable a patency rate as the conventional method even if it is used twice, both at the proximal and distal anastomoses, in autogenous vein grafting for arterial defects. Deformities of the inserted vessel should be minimized for patency by some means such as an application of fibrin glue so that the inserted vessel does not bulge under high arterial pressure. PMID- 7480241 TI - Delivery of demineralized bone powder by fibrin sealant. AB - The main purpose of this study was to determine whether the use of fibrin sealant as a delivery vehicle for demineralized bone powder would result in bone induction in heterotopic and orthotopic sites. Rat demineralized bone powder alone or in different concentrations of fibrin sealant matrix (4, 8, 15, and 45 mg/ml) was bioassayed for bone induction by implantation in intramuscular sites. Distribution of treatment groups was as follows: demineralized bone powder alone (n = 12), demineralized bone powder plus 4 mg/ml fibrin sealant (n = 11), demineralized bone powder plus 8 mg/ml fibrin sealant (n = 11), demineralized bone powder plus 15 mg/ml fibrin sealant (n = 11), demineralized bone powder plus 45 mg/ml fibrin sealant (n = 10), 4 mg/ml fibrin sealant (n = 13), and 45 mg/ml fibrin sealant (n = 11). In a second group of rats, 8-mm critical-sized calvarial defects were created and treated with demineralized bone powder plus 30 mg/ml fibrin sealant. Intramuscular implants were retrieved after 28 days, while calvarial implants were retrieved at 28 days (n = 8), 3 months (n = 8), or 4 months (n = 5). Implants were then x-rayed and submitted for histology. Results showed bone formation as evidenced by radiopacity and histology. Radiopacity measurements of demineralized bone powder implants alone or in a fibrin sealant matrix were associated with immature woven bone at the implantation site. Fibrin sealant allowed bone formation by demineralized bone powder to occur, improved the handling of demineralized bone powder, and facilitated the shaping of implants. PMID- 7480242 TI - No holds barred. PMID- 7480243 TI - The Plastic Surgery Educational Foundation, breast implants, and outcome studies. PMID- 7480244 TI - Combined dorsal forearm and lateral arm flap. AB - The free latissimus dorsi musculocutaneous flap and the free groin flap have been used for the coverage of medium- to large-sized soft-tissue defects in the hand. However, these are often too bulky for the hand, requiring secondary operation for thinning. We have used the dorsal forearm flap combined with the lateral arm flap for the coverage of large soft-tissue defects in the hand. This flap is based on the reversed vascular pedicle of the posterior interosseous artery. The posterior radial collateral branch of the profunda brachii artery is then anastomosed to the recipient vessel to augment the vascular supply to the lateral arm flap. We have used this combination flap in two clinical cases and achieved one-stage soft-tissue reconstruction of the hand. PMID- 7480245 TI - Paecilomyces variotii contamination in the lumen of a saline-filled breast implant. AB - This report describes a case of gross contamination with the filamentous fungus P. variotii cultured from an intraluminal saline breast implant removed from a patient 14 months after implantation because of severe capsular contracture. We suspect the fungal contamination occurred when a container of saline was left open in the operating room prior to filling and placement of the implant. This case may be the first documented report of microbial growth and reproduction in the internal environment of a saline implant. We assume that organisms such as P. variotii can survive--and accumulate biomass--on the minute amounts of substrates that diffuse across an implant envelope. PMID- 7480246 TI - Repair of radiation-induced vesicovaginal fistula with a rectus abdominis myocutaneous flap. AB - The cutaneous RAM flap was used successfully to repair a radiation-induced vesicovaginal fistula. Patient selection is important, ideally one with a fatty lower abdominal wall and without a previous incision transecting the inferior rectus muscle below the umbilicus. The RAM flap has excellent vascularity, allows augmentation of the bladder, and permits an extraperitoneal approach, factors that are important when operating in an irradiated field. PMID- 7480247 TI - Distraction lengthening of a replanted digit. AB - The Ilizarov technique was used to lengthen a replanted middle finger on a musician's left hand. Bone consolidation was achieved in 6 months without the need for a bone graft. The elongated digit has good sensibility, appropriate stability, and sufficient length, which were essential for a good performance in her profession. The unique indication to apply the callus distraction technique in such a special condition was described, and the relative merits of other treatment alternatives were discussed. PMID- 7480248 TI - Ganglion cyst presenting as a mass in the axilla. AB - A case is reported of an axillary mass that was consistent with a tumor. On exploration, the mass was found to be a large ganglion stemming from the shoulder joint. PMID- 7480249 TI - A simple fail-safe method for the harvesting of cranial bone. AB - The technique of cranial bone graft harvesting with the sagittal saw is described. With this technique regular bone grafts of any size can be obtained. Bone-graft size dictates the technique to be used. Small grafts (less than 3.0 cm in shortest dimension) are harvested in situ, while larger grafts are harvested by formal craniectomy. PMID- 7480250 TI - An alternative to a graft of skin in the treatment of lagophthalmos. AB - In spite of adequate preoperative evaluation, lagophthalmos may appear during a brow lift procedure if the patient has had a prior upper lid blepharoplasty. Relief of this condition may be obtained by incising the upper lid scar and spreading the wound. Subsequent granulation results in an acceptable scar. PMID- 7480251 TI - The double dermal keyhole pattern for breast reduction. AB - A small contribution is made to the dependable keyhole pattern in breast reduction. A double dermal modified keyhole closure is described. It is a simple, versatile, and adaptable method of closure regardless of the pedicle technique used for reduction mammaplasty. A triangular flap (skin and dermis) is incorporated on the medial keyhole flap. A deepithelialized dermal flap is created on the lateral keyhole flap. In the final closure of the wound, the medial triangular flap is sutured over the lateral deepithelialized dermal flap. A double dermal closure at the inverted T region is thereby created. The frequency of occurrence of wound spread and early postoperative wound dehiscence is decreased by this closure, which provides cosmetic improvement and wound support where it is needed the most. PMID- 7480252 TI - One rectus abdominis muscle for two separated soft-tissue reconstructions. AB - One rectus abdominis muscle flap, when based on superior and inferior pedicles, can be transferred simultaneously as two free-muscle flaps in two different configurations during the same operation. The two free flaps based on either pedicle are safe, reliable, and extremely well vascularized. Excellent functional and cosmetic results have been achieved in five patients with minimal donor-site morbidity. It has been our flap of choice for reconstructing the moderate- to large-sized defects at two separate sites of the upper and lower extremities. PMID- 7480253 TI - The split gluteus maximus muscle turnover flap. PMID- 7480254 TI - Therapeutic principles and options in radiation oncology. AB - Radiation therapy is used increasingly as a definitive or adjunctive treatment for cancer. Definitive irradiation allows functional preservation of structures critical to function. Adjunctive irradiation following surgical resection of the primary lesion treats regional lymph nodes and/or sites of primary extension at high risk for microscopic involvement. Specialized techniques can be applied to deliver higher total doses of irradiation and minimize late normal tissue effects to adjacent normal tissues. Local-regional tumor control combined with functional preservation represents the specific goals of radiation therapy applied alone or in conjunction with surgical resection. PMID- 7480255 TI - Choice of drugs after microsurgery. PMID- 7480256 TI - Implantation and expansion of split-thickness skin grafts. PMID- 7480257 TI - An occasion to revisit immediate tissue expansion. PMID- 7480258 TI - Grafting of bone from a dog into the human skull: an historical note. PMID- 7480259 TI - Atraumatic enophthalmos. PMID- 7480260 TI - A cheap imaging system. PMID- 7480261 TI - Vitiligo at incision lines. PMID- 7480262 TI - Utilization of the breast for penile reconstruction in a transsexual. PMID- 7480263 TI - Three helpful hints for face lift surgery. PMID- 7480265 TI - Removal of silicone from breast implants. PMID- 7480264 TI - Harvesting a full-thickness skin graft. PMID- 7480267 TI - Evaluating autogenous breast reconstruction with MRI. PMID- 7480266 TI - Removal of the septum and its replacement as a graft for the difficult nasal deformity. PMID- 7480268 TI - Editorials: for whom? PMID- 7480269 TI - The Oklahoma City bombing: a war of the worlds perspective. PMID- 7480270 TI - HLA typing in women with breast implants. AB - Since the 1970s, anecdotal reports have described a relatively small number of women who received silicone gel breast implants and later developed either a recognized rheumatologic disease or unexplained symptoms suggestive of an autoimmune disorder. The study reported here examined whether there is any association between the symptoms seen in implant patients and HLA molecules. One hundred and ninety-nine subjects were evaluated by HLA typing: symptomatic patients with implants (group I, n = 77), asymptomatic women with implants (group II, n = 37), healthy female volunteers without implants (group III, n = 54), and fibromyalgia patients without implants (group IV, n = 31). A statistically significant 68 percent of group I were positive for HLA-DR53, compared with 35 percent of group II and 52 percent of group III. The fibromyalgia patients were strikingly similar to group I women in terms of HLA-DR molecules, with 65 percent of group IV being positive for DR53. Group I also had a statistically significant increased frequency of HLA-DQ2. Asymptomatic women with implants (group II) had an increased frequency of DR1 and DQ1. In addition, 42 percent of symptomatic patients with implants formed autoantibodies to their own B cells; of these, 81 percent were DR53-positive. Although frequencies of capsular contracture and implant rupture were not significantly different in the two groups with implants, there were statistically significant associations in group I between contractures and ruptures and the presence of DR53 and B-cell autoantibodies. These data suggest that symptomatic patients with implants share important genetic characteristics (primarily HLA-DR53 positivity) that differentiate them from their asymptomatic counterparts. DR53 may be a marker of women who are predisposed by their HLA genotype to develop symptoms following exposure to silicone gel breast implants. PMID- 7480271 TI - The fate of breast implants: a critical analysis of complications and outcomes. AB - Complications and outcomes were monitored following the implantation of 1655 breast implants over a 15-year period. Smooth, polyurethane, and textured implants were used in a variety of clinical settings. The time course of capsular contracture was analyzed by the Kaplan-Meier method. Regardless of implant type or indication for surgery, the probability of contracture increased with time. Polyurethane-covered implants were associated with a significant reduction in the risk of contracture for at least 7 years following implantation. Smooth and textured silicone implants had contracture rates similar to each other, and the particular type of surface texturing (Biocel versus Siltex) was of no consequence. Contracture was more common following breast reconstruction and implant replacement than after augmentation mammaplasty and was not affected by filler material or implant size. Implant position did not alter the risk of contracture after augmentation; tissue expansion did not affect the risk of contracture after breast reconstruction. Infections were unusual but most common after reconstruction and unrelated to surface texture or filler material. Skin wrinkling was more frequent with saline implants and in the presence of surface texturing. Implant rupture was rare, with an incidence of 1 per 760 implant years. Implant-associated connective-tissue disease was noted in only one individual, an incidence of 1 per 3801 implant-years. PMID- 7480272 TI - Alveolar cleft bone grafts: results and imprecisions of the dental radiograph. AB - Alveolar cleft bone grafts customarily have been evaluated by one-dimensional dental radiographic measurements. Based on the dental radiograph, remarkable successes with just a single bone graft have been reported in the literature. At the Montreal Children's Hospital, the experience with 101 alveolar bone grafts in 62 cleft lip and palate patients was retrospectively reviewed to determine (1) the precision of dental radiographs at evaluating the clinical outcome, (2) the effect of dental maturation on alveolar bone grafts, and (3) the effect of augmentation bone grafts. The dental radiograph significantly overestimated the number of clefts that could be managed orthodontically (p < 0.01, McNemar's test) and thus poorly prognosticated the ability to achieve bony obliteration of the alveolar cleft and orthodontic closure of the dental gap. Bone grafts performed during the preeruptive canine dentition yielded significantly better results (p < 0.05, chi-squared test). With each subsequent augmentation bone-graft procedure performed, there existed a trend toward improved dental arch stability and radiographic and clinical outcomes. PMID- 7480273 TI - Secondary rhinoplasty following open rhinoplasty. AB - On the basis of 56 cases of secondary rhinoplasty following an initial open rhinoplasty, I conclude that secondary surgery is safe and effective. Either an open or closed approach can be used in most cases with little risk of skin necrosis or poor scar formation. The decision as to which approach to use depends on numerous factors. In general, a closed technique is favored when augmentation is the solution, while an open technique is favored when structural correction is required. Reopening the nose was done consistently without problems but was avoided in two patients because of severe thinning following previous defatting. However, certain "stigmata" were seen following open primary rhinoplasty and should be avoided: (1) a depressed, visible scar, (2) destruction of the soft tissue facets and nostril apices, (3) columella deformities with associated nostril asymmetry, and (4) excessive tip or supratip defatting. PMID- 7480274 TI - Influence of steroids on postoperative swelling after facialplasty: a prospective, randomized study. AB - Steroids are widely used in facial aesthetic surgery to reduce postoperative edema. We performed a randomized, double-blind study to try to document the effectiveness of this practice. Fifty consecutive facialplasty patients of one surgeon were randomized to steroid and no steroid groups. Steroid group patients received betamethasone 6 mg IM preoperatively. Postoperative scoring of swelling was performed at approximately days 5 and 9 by a single observer. There were no significant differences between the two groups at either postoperative interval or in the rate of improvement. Subgroups of patients who underwent additional procedures also showed no significant differences. We were not able to demonstrate any statistically significant difference in swelling after facialplasty with this steroid regimen. PMID- 7480276 TI - Orbicularis muscleplasty and face lift: a better orbital contour. AB - In the course of aging, the jugopalpebral tegumentary continuity disappears, and the orbital rim becomes more visible with the appearance of the palpebrojugal fold and the wrinkles of crow's feet. The deepening crease just above the sulcus cannot be well assessed because of the palpebral bags and the depression below the cheek due to ptosis of the premalar fat pad that comes down into the nasolabial fold. Repositioning of these volumes can be ensured by raising the cutaneous muscular and fat structures en bloc. Following an anatomic study of the aging process, I propose to section the fibers of the orbicularis muscle and to include them in the large cheek flap. This manipulation of the orbicularis will consolidate a weakened orbital septum. Resectioning of the fat pads under these conditions may not be necessary or at least may be more moderate because the deserted zones will be filled, which is what is desired. The cutaneous and capillary incisions adapted to the requirements of this technique are described. The results obtained with 123 patients over a period of 3 years by the same surgeon appear to be more complete and more natural because the transitional zones between the closed techniques of face lift on the one hand and of lower blepharoplasty on the other cease to exist. PMID- 7480275 TI - Subciliary malar augmentation and cheek advancement: a 6-year study in 22 patients undergoing blepharoplasty. AB - Twenty-two patients seeking aesthetic improvement of their eyelid and orbital cheek region underwent malar augmentation and cheek advancement in conjunction with lower eyelid blepharoplasty by means of a subciliary incision. The follow-up ranged from 12 to 78 months, with a mean follow-up of 44 months. Concomitant other aesthetic procedures were performed in 27 percent of the patients. Satisfactory goals of lower eyelid aesthetic improvement and malar augmentation with cheek advancement were achieved in 21 of 22 patients. A single patient had her implants removed within 3 weeks of her initial operation, at her request. Complications included hematoma formation in two patients, transient ectropion in two patients, and a palpable implant edge in a single patient. There were no long term functional or aesthetic sequelae. Malar augmentation and cheek advancement have been found to be a significant adjunct to lower eyelid blepharoplasty because they improve the appearance of the cheek region and add a youthful contour to the malar area. These straightforward procedures add an effective dimension to blepharoplasty in appropriately selected patients. PMID- 7480277 TI - Aesthetic restoration of the severely disfigured face in burn victims: a comprehensive strategy. AB - Although highly specialized burn centers have significantly reduced mortality rates following extensive total body surface area burns, survivors are often left with grotesque facial disfigurement. The strategy of modern facial restoration emphasizes enhancement of aesthetic appearance as significantly as mitigation of functional impairment. Criteria for success are (1) an undistracted "normal" look at conversational distance, (2) facial balance and symmetry, (3) distinct aesthetic units fused by inconspicuous scars, (4) "doughy" skin texture appropriate for corrective makeup, and (5) dynamic facial expression. Since 1985, the author has successfully restored 17 severely disfigured burned faces by replacement of entire aesthetic units with microvascular "prepatterned" composite flaps blended into the facial canvas by cosmetic camouflage techniques. The series includes hemiface (2), neck/jaw (5), chin/lower lip (3), cheek/malar (5), peri-orbital (2), nose (3), upper lip (4), and ear reconstructions (4). Flaps represented are free preauricular (1), radial forearm (6), ulnar forearm (1), free scapular (6), ilio-osteocutaneous (2), temporoparietal (8), vascularized forehead island (3), supraclavicular (1), and SMAS (1). Important to outcome is extensive initial intraoperative "sculpting" to simulate normal planes and contours. Seams are placed at junctions of facial components. Three-dimensional imaging is used to assess architectural asymmetries, and bone grafts are aided by computer-generated acrylic models. Adjunctive procedures include tensor fasciae latae slings, intraoperative tissue expansion, suction-assisted lipectomy, and scar management. After restoration of facial form and texture, flesh color make up and/or tattooing of beard, lips, scars, eyebrows, etc., aid to hide scars and pigment the skin to harmonize with the rest of the face. In all cases, facial integrity has been aesthetically restored and, in most instances, with makeup, is near normal in social settings at conversational distances. Facial animation is retained and color matches are excellent. One flap was lost early in the series. PMID- 7480278 TI - Maxillofacial growth after neck burn injury at a young age: an experimental study in the rabbit. AB - An experimental model was designed to define alterations in the normal mandibular growth process under the influence of postburn neck contractures. Additionally, this craniofacial growth model was used to compare two early treatment modalities of neck burns in their capability to minimize contracture and hence allow for normal mandibular growth and development. Growth implies increase in size as well as change in shape and position. These three aspects of growth were defined accordingly to Bjork as rotations. The intramatrix rotation expresses the change in mandibular shape, and the matrix rotation expresses the change in mandibular position relative to surrounding structures. The total rotation expresses both, and together with the measurement of the mandibular length, they represent the increase in mandibular size. Thirty-two 7-week-old rabbits were divided at random in four groups of eight rabbits each and randomized for selection for the 14 operation days defined as t = 0: Group A: controls to define normal mandibular growth Group B: untreated third-degree neck burns Group C: third-degree neck burns treated by a full-thickness skin graft Group D: third-degree neck burns treated by a myocutaneous flap All animals underwent placement of two bone markers in the maxilla. With biweekly intervals, standardized lateral skull roentgenographs were taken until the rabbits reached the age of 21 weeks. In this time period, major growth accelerations including the pubertal growth spurt took place. By the use of 13 reference points and 7 reference lines, rotations and distances were calculated. Statistical analysis of the data was performed. The results show that the normal mandibular length was unaffected in all groups. There were no statistically significant changes in matrix, intramatrix, and total rotations of the mandible and the maxilla. There was a statistically significant difference in the displacement of the mandibular reference point between all groups, suggesting a variable degree of normal backward skull rotation, namely, due to group B. Explanations to be considered concerning the fact that the only minor differences were found in group B: 1. Drawbacks of the animal model: differences in skin texture, postnatal mandibular growth, and head position compared with those of humans. 2. Other functional adaptation mechanisms such as changes in head position, which are recruited at first in adapting to disturbances of homeostasis, were not measured. Soft-tissue compensation probably has overcome major bony deformations. Nevertheless, some drawbacks of the model can be viewed as ideal concerning treatment of neck burns.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7480279 TI - The application of skin external expander to postburn advanced scar contracture. AB - In recent years, the universally used internal expansion by implanting a silicone elastomer balloon into the soft tissue under the skin has shown certain disadvantages, i.e., certain complications associated with the operation, high cost, and the fact that the materials cannot be recycled. That the materials are used only once adds to the operating cost. In view of these disadvantages, we have designed and developed a kind of mechanical external skin expander, which is composed of (1) an adjustable tractor, (2) preserving splints, and (3) a traction frame. The tractor is used to grip the contracted skin in order to make the soft tissue of the skin expand with the lapse of time and increase in traction weight. The external skin expansion has the following advantages compared with internal expansion: avoiding the complications caused by implanting the soft-tissue expander beneath the skin; simplicity and efficiency of operation; and the fact that the equipment can be reused, reducing the operating which is suitable for these basic units. The data of the expanded skin area can be calculated through the computer. Since June of 1993, a satisfactory curative effect has been obtained in 44 patients suffering from advanced postburn scars. Thirty-two patients among them were treated for more than half a year, with the scar becoming normal in both appearance and function. External skin expansion provides a new method for expansion of skin soft tissue. It has a bright future in its clinical application. PMID- 7480280 TI - Latissimus dorsi musculocutaneous flap without muscle. AB - The possibility of raising the cutaneous island of the latissimus dorsi musculocutaneous flap without muscle based on only one cutaneous perforator is presented in this paper. An anatomic study performed in 40 fresh cadaver specimens injected with colored latex demonstrated that the vertical intramuscular branch of the thoracodorsal artery gives off two to three cutaneous branches (perforators) that are consistently present. The largest one, measuring approximately 0.4 to 0.6 mm in diameter, provides the blood supply to a 25 x 15 cm cutaneous island. The incorporation of the proximal trunk of the thoracodorsal artery lengthens the pedicle, facilitating the anastomosis or the arc of rotation (in the case of island flaps) but does not increase the amount of tissue transferable. Five clinical cases were done with this technique without tissue necrosis or flap loss. PMID- 7480281 TI - TRAM flap vascular delay for high-risk breast reconstruction. AB - The purpose of this study was to evaluate the intraoperative changes in physiologic blood pressure following vascular delay of the TRAM flap. Ligation of the superficial and deep inferior epigastric vessels 2 weeks prior to the TRAM flap was performed. The incidence of fat necrosis was 4.3 percent in 23 high-risk patients who underwent 30 immediate breast reconstructions. Direct measurement of blood pressure in the deep inferior epigastric artery and vein was performed in a control group without delay consisting of 13 low-risk patients and in the study group of 7 high-risk patients who underwent vascular delay. Changes in TRAM flap perfusion pressure were examined following the change in location of the flap from the abdomen to the chest. Blood pressure measurements demonstrated that arterial pressure in the proximal stump of the deep inferior epigastric artery was 64 percent in the study group prior to delay and 72 percent in the control group. The physiologic response to vascular delay included an overall increase in arterial pressure with a decrease in venous congestion. TRAM flap perfusion pressure was significantly increased from 13.3 mmHg (control) to 40.3 mmHg (delayed) in the region of the midrectus perforators (p < 0.05). These data suggest that the technique of TRAM flap delay may increase the reliable tissue volume and improve the safety of the TRAM flap. PMID- 7480282 TI - Quantification of functional deficits associated with rectus abdominis muscle flaps. AB - To evaluate the deficits in trunk flexion due to partial or complete unilateral rectus abdominis muscle flap transfer in reconstructive surgery, 27 patients (11 females, 16 males) with rectus abdominis muscle flap transfer dating back at least 3 years, clinically healthy at the time of testing, and 22 controls (10 females and 12 males), comparable in age and body mass index, were compared. For clinical assessment of power of the straight and oblique abdominal muscles, Janda's test of muscle function was used, and torque (in N.m) of isometric trunk flexion at 0, 5, 10, 20, and 30 degrees was determined with the TEF Modular Component, an auxiliary unit of the Cybex 6000. Results were analyzed for males and females separately. Relative torque (N.m/kg) of operated male and female patients was significantly lower (p < 0.05) as compared with controls. Differences were more pronounced in males than in females. Clinically, there were no significant between-group differences in the power of the straight and oblique abdominal muscles. In conclusion, trunk flexion deficits following rectus abdominis muscle flap transfer can be compensated for satisfactorily in most cases, supporting the use of this muscle in reconstructive surgery, if patients are selected carefully. PMID- 7480283 TI - A true osteomyocutaneous free-flap model in rats: the saphenous artery osteomyocutaneous flap. AB - A new composite free-flap model, namely, the saphenous artery osteomyocutaneous flap, is described in the rat. This is a true osteomyocutaneous flap composed of a skin island from the medial aspect of the lower leg, the gracilis and semitendinosus muscles, and a bone segment from the tibia based on the saphenous vascular pedicle. After anatomic studies in 10 rats, 20 flaps were transplanted microsurgically to the abdominal region in the recipient rats with a 90 percent success rate. In selected animals, arteriography was carried out. The results from gross observation of the flap survival and histologic and fluorochrome bone labeling studies revealed a complete survival for each component of the flap in all animals with a patent vascular pedicle at 2 weeks after transfer. The model seems to be suitable for use in metabolic, vascular, and immunologic experimental studies on composite free flaps. PMID- 7480284 TI - Assessment of microcirculation of an axial skin flap using indocyanine green fluorescence angiography. AB - In many cases the complexities of skin-flap microcirculation are difficult to assess despite all the subjective and objective examination techniques available today. Adequate microcirculation is essential for tissue viability, so any method employed for studying microcirculation should provide as accurate an assessment of the prevailing conditions as possible. Of all the clinical methods, the fluorescence technique using the dye sodium fluorescein has so far provided the most reliable results. However, the pharmacokinetic properties of this tracer have prevented the technique from becoming established in clinical practice. The fluorescent dye indocyanine green (Cardio Green), on the other hand, has far more favorable pharmacokinetics. In an experimental animal model, the fluorescence technique using indocyanine green (indocyanine green angiography, ICGA) was used to study postoperative changes in the microcirculation of a skin flap. On the day of operation, indocyanine green angiography revealed a state of hemodynamic imbalance for which the organism was able to compensate in the postoperative phase with the aid of humoral, physical, and metabolic factors. With indocyanine green angiography it was possible to quantify objectively the new hemodynamic equilibrium. Basically, microcirculation may be quantified in temporal and spatial terms. The significant objectivity of indocyanine green angiography and short intervals between each examination favor its possible and meaningful use in clinical practice and give cause for continuing studies. PMID- 7480285 TI - Effects of delay procedures on vasculature and survival of arterialized venous flaps: an experimental study in rabbits. AB - The effect of delay procedures on the survival of large arterialized venous flaps was studied in a rabbit ear model. Both ears of 25 New Zealand White rabbits (n = 50) were randomized into three operative groups receiving (1) no delay, (2) limited delay, or (3) extensive delay. Two weeks later the ear flap was completely divided, and arterialization of the flap venous tree was achieved by anastomosing the central artery of the ear to the anterior branch of the flap central vein. Survival area of 10 flaps in each group was assessed at 14 days. Flap vasculature was assessed in 10 additional ears in the nondelayed and extensively delayed groups by standardized vascular injections with radiopaque silicon rubber immediately, 6 hours, 24 hours, and 2 weeks after arterialization. Delay procedures significantly increased (p = 0.001, unpaired two-tailed Student's t test) survival of arterialized venous flaps in this model. Whereas all flaps in the non-delayed group underwent total necrosis, the mean viable surface area of the flaps was 67.9 percent in the limited delay group and 94.0 percent in the extensively delayed group. Lastly, angiographic studies suggest that delay procedures prior to arterialization of the venous tree enhance perfusion of the arterial tree by increasing venoarterial communications. PMID- 7480286 TI - Complete elimination of large cafe-au-lait birthmarks by the 510-nm pulsed dye laser. AB - Traditional treatment modalities for cafe-au-lait spots include cryotherapy, dermabrasion, and excision with or without skin grafting. These treatments have shown varying degrees of success and, in some cases, have yielded significant side effects, such as permanent pigmentary alteration and scarring. The 510-nm pulsed dye laser has been shown to selectively destroy melanosomes that make up such epidermal pigmented lesions as cafe-au-lait spots without significant adverse sequelae. Thirty patients with 34 large cafe-au-lait birthmarks received 510-nm pulsed dye laser treatments at 6- to 8-week intervals with energy densities ranging from 2.0 to 4.0 J/cm2. An average of 8.4 laser treatments were necessary to achieve complete elimination of the cafe-au-lait marks under study. No lesional recurrences have been observed 1 year following termination of treatment. PMID- 7480287 TI - Utilizing the osseointegration principle for fixation of nail prostheses. AB - A new osseointegrated anchorage device that holds a prosthetic nail plate was used successfully in eight patients, for a total of nine missing fingernails, secondary to trauma. There was only one early failure associated with patient noncompliance. The average follow-up time was 18 months. Details of the technique and three illustrative cases are presented. PMID- 7480288 TI - Effect of motion on digital nerve repair in a fresh cadaver model. AB - Information in the literature regarding the postoperative management of digital nerve lacerations is vague, and postoperative immobilization for up to 3 weeks is frequently recommended. In order to define more precisely what, if any, postoperative restrictions are necessary, a fresh cadaver model was designed for digital nerve division, resection, repair, and passive motion. Ten digital nerves were divided at the proximal interphalangeal joint and then repaired, mobilized, and inspected. Intact nerve repairs were serially resected in order to determine the limits of resection that would allow motion without repair disruption. All repairs were resistant to disruption even with hyperextension up to a resection length of 2.5 mm, and all repairs were resistant to disruption if splinted in neutral up to a resection length of 5 mm. There was not 100 percent disruption of repairs until a resection length of 1 cm and range of motion including hyperextension. These results give valuable objective data that can be used to guide early motion and splinting protocols after various degrees of digital nerve injury and repair. PMID- 7480289 TI - An experimental model to determine the effect of irradiated tissue on neutrophil function. AB - Complications of irradiated tissue include infections and impaired healing. Although fibrosis and hypovascularity contribute, a cellular mechanism has not been identified. This study examines the effect of radiation (10 to 30 Gy) on neutrophil function in a rabbit wound cylinder model. At 3 to 12 weeks after radiation, subcutaneous wound cylinders were implanted in both irradiated and control fields in 19 rabbits. Wound neutrophils were subsequently assayed for phagocytosis (3H-labeled Staphylococcus aureus assay), superoxide production (cytochrome c reduction assay), and surface Mac-1 expression (flow cytometric assay using MHM 23 monoclonal antibody). Phagocytosis of 3H-labeled S. aureus was significantly lower in neutrophils from irradiated fields compared with controls at 6 and 12 weeks after radiation (6.5 versus 18.9 bacteria per neutrophil at 12 weeks; p = 0.027). Stimulated neutrophils from irradiated tissue could not increase superoxide production or Mac-1 expression as much as controls, with differences increasing as postirradiation time increased. The diminished phagocytosis, superoxide production, and Mac-1 expression provide a cellular mechanism that may account for susceptibility to infection and poor healing in irradiated tissues. PMID- 7480290 TI - A rapid in vitro assay of cellular chemomigration in an epithelial carcinoma cell line. AB - We have studied the chemomigration activity of an epithelial carcinoma cell line using a modified 96-well Boyden chamber apparatus consisting of upper and lower wells separated by an 8-microns pore polycarbonate filter. Cells from the malignant squamous carcinoma cell line A-431 were plated in the upper wells over a collagen IV-coated filter. In chemokinesis assays, the cells were allowed to migrate toward NIH 3T3 fibroblast-conditioned medium or control media in the lower wells for 6 hours at 37 degrees C with 10% CO2. A-431 cells preferentially migrate across the barrier toward conditioned media but not control media. Control normal keratinocytes showed no migration. A highly metastatic melanoma cell line and poorly metastatic melanoma cell line, in which chemomigration has been shown previously to correlate with metastatic potential, were used as positive and negative cellular controls. This system provides a rapidly quantifiable method by which the invasion characteristics of multiple cell lines can be studied simultaneously in a single assay using the 96-well format. PMID- 7480291 TI - Quo vadis. PMID- 7480293 TI - Salvage of an amputated ear temporarily lodged in a forearm. AB - After amputation of an ear, the best results are attained through the use of microvascular replantation. The literature reports very few examples of the successful replantation of a whole ear when compared with the vast number of attempts. We report the case of a 17-year-old boy with a recurrence of a voluminous arteriovenous malformation in the right temporoparietal area. Since radical excision was impossible without removing the ear, the cartilage with the perichondrium was inserted into the distal third of the left forearm. Six months later a composite free flap formed by a radial forearm flap including the cartilage was transferred to the permanent site. In the following months, the edema of the flap obscured the outline of the ear, forcing us to remove the skin and subcutaneous layer of the flap and cover the anterior face of the ear with a split-thickness skin graft. The indications and advantages of such a procedure are also discussed. PMID- 7480292 TI - Managed care: now you see it, now you don't. PMID- 7480295 TI - Seromuscular patch as an adjunct in reconstruction of the cervical esophagus with free jejunal transfer. AB - During cervical esophageal reconstruction, an auxiliary piece of seromuscular tissue can be developed based on the same vascular pedicle as the interpositional piece. This segment of vascularized tissue can be used as a reconstructive aid in compromised areas where wound problems may create disastrous complications (e.g., carotid rupture or tracheoesophageal fistula). The proximal bowel anastomosis can be reinforced, an exposed carotid can be covered, or the distal jejunal anastomosis can be circumferentially wrapped. Any skin deficiencies created by placement of the seromuscular sleeve are easily corrected by a split-thickness skin graft. PMID- 7480294 TI - Toxic shock syndrome as a complication of breast prostheses. AB - A case of a 21-year-old woman who developed toxic shock syndrome 6 days after augmentation mammaplasty with saline breast implants is reported. The infecting organism was S. aureus that was toxic shock syndrome exotoxin 1-negative and staphylococcal endotoxin B-positive. The causes and etiology of this rare postoperative complication are discussed. PMID- 7480296 TI - A new skin suture technique for multiple Z-plasty. AB - A new skin suture technique was developed for multiple Z-plasty. In this technique, the wound of multiple Z-plasty is closed by several continuous locking sutures. With use of this technique, the operative time is reduced, and suture removal is facilitated to alleviate pain and discomfort on the part of the patient. PMID- 7480297 TI - Prevention of hypertrophic scars by long-term paper tape application. AB - Longitudinal stretching of wounds that cross the relaxed skin tension lines appears to be the stimulus that causes the body to form hypertrophic scars. Control or elimination of such stretching forces by the long-term use of paper tape, beginning at 2 weeks, has been effective in preventing such hypertrophic scarring. PMID- 7480298 TI - A composite lateral arm flap for the secondary repair of a multiply ruptured Achilles tendon. AB - Reported uses of the lateral arm flap for Achilles tendon reconstruction are few, and all refer to delayed primary repair. We would not only like to suggest its use for secondary repairs but also to recommend the incorporation of a suitable olecranon fragment with the triceps tendon graft as a workable anchoring device. This provides a more secure insetting in the all too common operative situation of a short, frayed distal stump and, most important, helps to insure against postoperative rerupture. PMID- 7480299 TI - A second look at the second-look technique in face lifts. AB - Hematoma remains a problematic complication of face lift surgery. Sequelae of hematomas may include temporary skin irregularities, a prolonged postoperative recovery period, skin necrosis, and a compromised final result. Attempts to maximize intraoperative and postoperative hemostasis and thus prevent postoperative hematoma formation have focused on the use of local anesthetics with epinephrine modifications, topical thrombin, bed rest, blood pressure control, local anesthetics with intravenous sedation rather than general anesthesia, and the use of the "second-look" technique popularized 30 years ago. When initially practiced by the senior author, the second-look technique resulted in a dramatic decrease in the incidence of hematoma formation from 21.7 to 6.7 percent. It was concluded at that time that the second-look technique was responsible for the reduced incidence of this face lift complication. However, the present series of 82 face lifts performed using a modification of the original Marcaine/epinephrine infiltration technique without the second-look technique demonstrates an overall incidence of hematoma formation of 7.3 percent. These results demonstrate no advantage of the second-look technique over the traditional technique when epinephrine is infiltrated in the fashion described and meticulous hemostasis is obtained. PMID- 7480300 TI - Plastic surgery at the Johns Hopkins Hospital in the 1940s: personal reflection on a period of transition. PMID- 7480301 TI - The sharp treatment of abrasion with dirt tattoo. PMID- 7480302 TI - Microskin grafts using skin-graft meshers. PMID- 7480303 TI - Frontal sinus obliteration. PMID- 7480304 TI - Sphincter pharyngoplasty. PMID- 7480305 TI - Intraductal migration of silicone. PMID- 7480306 TI - Silicone breakdown and clinical implications of mammary and extramammary synovial metaplasia in periprosthetic capsules. PMID- 7480309 TI - Salmonella infection following breast reconstruction. PMID- 7480307 TI - Localized atypical mycobacteria. PMID- 7480308 TI - Silicone gel implant explantation. PMID- 7480310 TI - Gigantic lipomas. PMID- 7480311 TI - Lateral paracanthal incision. PMID- 7480312 TI - Vaginal stents for McIndoe's vaginal construction. PMID- 7480313 TI - Cardiomyoplasty. PMID- 7480314 TI - Complications of harvesting cranial bone grafts. PMID- 7480315 TI - Surgical treatment of axillary osmidrosis. PMID- 7480317 TI - Treatment for self-inflicted laceration scars. PMID- 7480316 TI - Correction of gynecomastia with an inframammary incision and subsequent scar. PMID- 7480318 TI - Autotransfusion and reduction mammaplasty. PMID- 7480319 TI - Insect cell expression of recombinant imidazoleglycerolphosphate dehydratase of Arabidopsis and wheat and inhibition by triazole herbicides. AB - Imidazoleglycerolphosphate dehydratase (IGPD; EC 4.2.1.19), which is involved in the histidine biosynthetic pathway of Arabidopsis thaliana and wheat (Triticum aestivum), has been expressed in insect cells using the baculovirus expression vector system. N-terminal amino acid sequencing indicated that recombinant IGPDs (rIGPDs) were produced as mature forms via nonspecific proteolytic cleavages in the putative transit peptide region. The wheat rIGPD contained one Mn atom per subunit, and the Mn was involved in the assembly of the subunits to form active IGPDs. Protein-blotting analysis, using antibodies raised against the wheat rIGPD, indicated that IGPD was located in the chloroplasts of wheat. The rIGPDs of Arabidopsis and wheat, which were 86% identical in their primary structures deduced from the cDNAs, exhibited similar properties in terms of the molecular mass, pH optimum, and the Km for the substrate, imidazoleglycerolphosphate. However, the nonselective herbicides 3-amino-1,2,4-triazole and a newly synthesized triazole [(1R*, 3R*)-[3-hydroxy-3-(2H-[1,2,4]triazole-3-yl) cyclohexyl]- phosphonic acid], inhibited Arabidopsis and wheat IGPDs in a mixed type and a competitive manner, respectively. PMID- 7480320 TI - Light-dependent and tissue-specific expression of the H-protein of the glycine decarboxylase complex. AB - Glycine decarboxylase is a mitochondrial enzyme complex, which is the site of photorespiratory CO2 and NH3 release. Although the proteins that constitute the complex are located within the mitochondria, because of their intimate association with photosynthesis their expression is controlled by light. Comparisons of the kinetics of mRNA accumulation between the small subunit of ribulose-1,5-bisphosphate carboxylase/oxygenase and the H-protein of glycine decarboxylase during the greening of etiolated Arabidopsis thaliana suggest that their expression is controlled in parallel. A genomic clone for the H-protein (gdcH) was isolated from Arabidopsis and sequenced. The upstream region from -856 to +62 was fused to the beta-glucuronidase (GUS) reporter gene, and this construct was transformed into tobacco. This 5' upstream regulatory region appears to control GUS expression in a manner very similar to that of the endogenous H-protein gene. Constructs with deletions in the 5' upstream region were transformed into tobacco. These deletions revealed that light-dependent and tissue-specific expression was largely controlled by a 259-bp region between -376 and -117 bp. This region contains several putative GT boxes with the GGTTAA consensus core sequence. Once these strong light-dependent elements were removed, a second level of control was revealed. In constructs in which the gdcH 5' regulatory region was shortened to -117 bp or less, there was more GUS activity in the roots than in the leaves, and in dark-grown plants than in light-grown plants. This suggests that more proximal control elements may be responsible for the constitutive low levels of gene expression noted in all nonphotosynthetic tissues. PMID- 7480321 TI - Retention of cadmium in roots of maize seedlings. Role of complexation by phytochelatins and related thiol peptides. AB - Cd from roots of maize was partitioned in seedlings exposed to 3 microM CdSO4 for 1 to 7 d. Most of the root Cd (92-94%) was buffer soluble and provided the classical metal-induced cysteine-rich, high-molecular-weight Cd-binding complex. This complex, however, bound only part of the Cd within the roots, from 19% after 1 d of exposure to 59% by d 7. Three families of peptides formed the Cd-binding complex: (gamma-glutamic acid-cysteine)n-glycine [(gamma-Glu-Cys)n-Gly], or phytochelatins, (gamma-Glu-Cys)n, and (gamma-Glu-Cys)n-Glu. The monothiols gamma Glu-Cys-Gly (glutathione), gamma-Glu-Cys, and gamma-Glu-Cys-Glu were absent from the complex. The n2 oligomers of any peptide were the least concentrated, whereas the n3 and n4 oligomers increased in the complex with exposure to Cd. By d 7, 75% of (gamma-Glu-Cys)4-Gly, 80% of (gamma-Glu-Cys)4, and 73% of (gamma-Glu-Cys)3-Glu were complexed with Cd. The peptide thiol:Cd molar ratio for the complexes was 1.01 +/- 0.07, as if the minimal amount of thiol was used to bind Cd. Acid-labile sulfide occurred in the complexes from d 1 onward at the low S2-;Cd molar ratio of 0.18 +/- 0.02. PMID- 7480322 TI - Differential accumulation of antioxidant mRNAs in Arabidopsis thaliana exposed to ozone. AB - Antioxidant isoenzymes function to eliminate free radicals and are localized to several different subcellular compartments within the plant cell. In Arabidopsis thaliana exposed to ozone (O3), we have monitored the accumulation of mRNAs encoding both cytosolic and chloroplastic antioxidant isoenzymes. Two different O3 exposure protocols yielded similar results. Upon O3 exposure, the steady-state levels of three mRNAs encoding cytosolic antioxidant isoenzymes (ascorbate peroxidase, copper/zinc superoxide dismutase, and glutathione S-transferase) increase. The glutathione S-transferase mRNA responds very quickly to the oxidative stress (2-fold increase in 30 min) and is elevated to very high levels, especially in plants grown with a 16-h photoperiod. In contrast, O3 exposure causes a decline in the levels of two chloroplastic antioxidant mRNAs (iron superoxide dismutase and glutathione reductase) and two photosynthetic protein mRNAs (chlorophyll a/b-binding protein and ribulose-1,5-bisphosphate carboxylase/oxygenase small subunit). We show that this decline does not include all mRNAs encoding chloroplast-targeted proteins, since O3 causes an elevation of mRNA encoding the chloroplast-localized tryptophan biosynthetic enzyme phosphoribosylanthranilate transferase. Two alternative hypotheses that could explain this differential mRNA accumulation in response to O3 are discussed. PMID- 7480323 TI - Specific binding of a dichloroacetamide herbicide safener in maize at a site that also binds thiocarbamate and chloroacetanilide herbicides. AB - Dichloroacetamide safeners such as N,N-diallyl-2,2-dichloroacetamide and (R,S)-3 dichloroacetyl-2,2,5-trimethyl-1,3-oxazolidine protect maize (Zea mays) against injury from thiocarbamate and chloroacetanilide herbicides. Binding activity of tritium-labeled (R,S)-3-dichloroacetyl-2,2,5-trimethyl-1,3-oxazolidine (15 Ci/mmol; referred to as [3H]Saf) was characterized in extracts of etiolated maize seedlings. The binding is saturable, involves a single class of binding sites (Kd 0.12 microM; maximal binding in coleoptiles 0.53 nmol/g fresh weight, equivalent to 55 pmol/mg protein), and is sensitive to boiling and protease treatment. Binding in etiolated maize seedlings is highest in the coleoptile and lowest in the leaves. Binding of [3H]Saf also occurs in etiolated sorghum (Sorghum bicolor) shoots but not several other cereals. There is a good correlation between known safener effectiveness and the concentration that inhibits [3H]Saf binding half maximally among 21 dichloroacetamides and related compounds. N,N-Diallyl-2,2 dichloroacetamide had the lowest inhibitor concentration that reduces specific binding by 50% (IC50), 0.01 microM. [3H]Saf binding is inhibited by 4 chloroacetanilide herbicides with IC50 values of 0.07 to 0.48 microM and by 12 thiocarbamate herbicides and analogs with IC50 values of 0.06 to 2.3 microM. The inhibition of [3H]Saf binding by alachlor and S-ethyl dipropylthiocarbamate is competitive. PMID- 7480325 TI - Cloning and characterization of a glutathione S-transferase that can be photolabeled with 5-azido-indole-3-acetic acid. AB - Previously, we identified a soluble protein from Hyoscyamus muticus that was photolabeled by 5-azido-indole-3-acetic acid. This protein was determined to be a glutathione S-transferase (GST; J. Bilang, H. Macdonald, P.J. King, and A. Sturm [1993] Plant Physiol 102: 29-34). We have examined the effect of auxin on the activity of this H. muticus GST. Auxins reduced enzyme activity only at high concentrations, with 2,4-dichlorophenoxyacetic acid (2,4-D) and 2,4,5 trichlorophenoxyacetic acid being more effective than indole-3-acetic acid (IAA) and naphthylacetic acid. IAA was a noncompetitive inhibitor, whereas inhibition by 2,4-D was competitive with respect to 1-chloro-2,4-dinitro-benzene. We also present the sequence of a full-length cDNA clone that codes for a GST and contains all partial amino acid sequences of the purified protein. The auxin binding GST was found in high amounts in roots and stems and low amounts in leaves and flower buds. The steady-state mRNA level was not regulated by IAA or naphthylacetic acid, whereas 2,4-D and 2,3-dichlorophenoxyacetic acid increased mRNA levels. We propose a model in which 2,4-D is a substrate for GST, whereas IAA binds at a second site, known as a ligandin-binding site for the purpose of intracellular transport. PMID- 7480324 TI - Adenosine 5'-diphosphate-glucose pyrophosphorylase from potato tuber. Significance of the N terminus of the small subunit for catalytic properties and heat stability. AB - cDNAs encoding the large subunit and a possibly truncated small subunit of the potato tuber (Solanum tuberosum L.) adenosine 5'-diphosphate-glucose pyrophosphorylase have been expressed in Escherichia coli (A.A. Iglesias, G.F. Barry, C. Meyer, L. Bloksberg, P.A. Nakata, T. Greene, M.J. Laughlin, T.W. Okita, G.M. Kishore, J. Preiss, J Biol Chem [1993] 268: 1081-1086). However, some properties of the transgenic enzyme were different from those reported for the enzyme from potato tuber. In this work, extension of the cDNA was performed to elongate the N terminus of the truncated small subunit by 10 amino acids. This extension is based on the almost complete conservation seen at the N-terminal sequence for the potato tuber and the spinach leaf small subunits. Expressing the extended cDNA in E. coli along with the large subunit cDNA yielded a transgenic heterotetrameric enzyme with similar properties to the purified potato tuber enzyme. It was also found that the extended small subunit expressed by itself exhibited high enzyme activity, with lower affinity for activator 3 phosphoglycerate and higher sensitivity toward inorganic phosphate inhibition. It is proposed that a major function of the large subunit of adenosine 5' diphosphate-glucose pyrophosphorylases from higher plants is to modulate the regulatory properties of the native heterotetrameric enzyme, and the small subunit's major function is catalysis. PMID- 7480326 TI - N-acylphosphatidylethanolamine in dry and imbibing cottonseeds. Amounts, molecular species, and enzymatic synthesis. AB - N-Acylphosphatidylethanolamine (NAPE), an unusual acylated derivative of phosphatidylethanolamine (PE), was recently shown to be synthesized from PE and free fatty acids in cotyledons of cotton (Gossypium hirsutum L.) seedlings (K.D. Chapman, T.S. Moore [1993] Plant Physiol 102: 761-769). Here we report that NAPE is present in dry seeds of cotton and increases with time of imbibition from 2.31 nmol/seed in dry seeds to 4.26 nmol/seed in 4-h-soaked seeds. Total phospholipid/seed also increased such that the relative percentage of NAPE was similar in dry and soaked seeds (2.3 mol% compared to 2.6 mol%, respectively). The major molecular species of NAPE were identified in both dry and soaked seeds by fast atom bombardment mass spectrometry and collisionally activated dissociation tandem mass spectrometry as 16:0/18:2-PE(N-palmitoyl), 16:0/18:2 PE(N-linoleoyl), and 18:2/18:2-PE(N-palmitoyl). The specific activity of NAPE synthase in seed extracts increased with increasing time of imbibition from 35 pmol h-1 mg-1 protein in dry seeds to 129 pmol h-1 mg-1 protein in 4-h-soaked seeds. Collectively, our results indicate that NAPE is present in dry cottonseeds and synthesized during imbibition. The biosynthesis of NAPE provides a mechanism for maintaining membrane integrity during seed rehydration and may indicate that NAPE plays a protective role in intracellular membranes of plant tissues, as has been suggested for intracellular membranes of animal tissues. PMID- 7480327 TI - The diageotropica mutation and synthetic auxins differentially affect the expression of auxin-regulated genes in tomato. AB - The effect of a tomato (Lycopersicon esculentum) mutation, diageotropica (dgt), on the accumulation of mRNA corresponding to tomato homologs of three auxin regulated genes, LeAux, LeSAUR, and Lepar, was examined. The dgt mutation inhibited the induction of LeAux and LeSAUR mRNA accumulation by naphthalene acetic acid (NAA) but had no effect on NAA-induced Lepar mRNA accumulation. The effect of two synthetic auxins, NAA and 3,7-dichloro-8-quinoline carboxylic acid (quinclorac), on the accumulation of LeAux, LeSAUR, and Lepar mRNA was also examined. Quinclorac induced the expression of each of the auxin-regulated genes, confirming its proposed mode of herbicidal action as an auxin-type herbicide. Concentrations of quinclorac at least 100-fold higher than NAA were required to induce LeAux and LeSAUR mRNA accumulation to similar levels, whereas Lepar mRNA accumulation was induced by similar concentrations of NAA and quinclorac. Collectively, these data suggest the presence of two auxin-dependent signal transduction pathways: one that regulates LeSAUR and LeAux mRNA accumulation and is interrupted by the dgt mutation and a second that regulates Lepar mRNA accumulation and is not defective in dgt tomato hypocotyls. These two auxin regulated signal transduction pathways can be further discriminated by the action of two synthetic auxins, NAA and quinclorac. PMID- 7480328 TI - Temporal and spatial expression of amygdalin hydrolase and (R)-(+)-mandelonitrile lyase in black cherry seeds. AB - In black cherry (Prunus serotina Ehrh.) macerates, the cyanogenic diglucoside (R) amygdalin undergoes stepwise degradation to HCN catalyzed by amygdalin hydrolase (AH), prunasin hydrolase, and (R)-(+)-mandelonitrile lyase (MDL). A near full length AH cDNA clone (pAH1), whose insert encodes the isozyme AH I, has been isolated and sequenced. AH I exhibits several features characteristic of beta glucosidases of the BGA family, including their likely nucleophile center (isoleucine-threonine-glutamic acid-asparagine-glycine) and acid catalyst (asparagine-glutamic acid-proline/isoleucine) motifs. The temporal expression of AH and MDL in ripening fruit was analyzed by northern blotting. Neither mRNA was detectable until approximately 40 days after flowering (DAF), when embryos first became visible to the naked eye. Both mRNAs peaked at approximately 49 DAF before declining to negligible levels when the fruit matured (82 DAF). Taken together with enzyme activity data, these time courses suggest that AH and MDL expression may be under transcriptional control during fruit maturation. In situ hybridization analysis indicated that AH transcripts are restricted to the procambium, whereas MDL transcripts are localized within cotyledonary parenchyma cells. These tissue-specific distributions are consistent with the major locations of AH and MDL protein in mature seeds previously determined by immunocytochemistry (E. Swain, C.P. Li, and J.E. Poulton [1992] Plant Physiol 100:291-300). PMID- 7480329 TI - Evidence that plant K+ channel proteins have two different types of subunits. AB - Plant K+ channel proteins have been previously characterized as tetramers of membrane-spanning alpha subunit polypeptides. Recent studies have identified a 39 kD, hydrophilic polypeptide that is a structural component of purified animal K+ channel proteins. We have cloned and sequenced an Arabidopsis thaliana cDNA encoding a 38.4-kD polypeptide that has a sequence homologous to the animal K+ channel beta subunit. Southern and northern analyses indicate the presence of a gene encoding this cDNA in the Arabidopsis genome and that its transcription product is present in Arabidopsis cells. To our knowledge, this is the first report to document the presence of K+ channel beta subunits in plants. PMID- 7480330 TI - The electronic Plant Gene Register. PMID- 7480331 TI - Stress activation of a bean hydroxyproline-rich glycoprotein promoter is superimposed on a pattern of tissue-specific developmental expression. AB - The HRGP4.1 gene, which encodes a cell wall hydroxyproline-rich glycoprotein, was isolated from a genomic library of bean (Phaseolus vulgaris L.). Two transcripts, one induced by wounding and one by elicitation, were transcribed from the same initiation site. The gene encodes a polypeptide of 580 amino acids with the amino terminal half consisting of repeats of the sequence serine-(proline)4-lysine histidine-serine-(proline)4-(tyrosine)3-histidi ne and the carboxyl-terminal half composed of repeats of the sequence serine-(proline)4-valine-tyrosine-lysine tyrosine-lysine. A 964-bp upstream promoter fragment was translationally fused to the beta-glucuronidase reporter gene (Escherichia coli uidA) and transferred into tobacco by Agrobacterium tumefaciens-mediated leaf disc transformation. Analysis of beta-glucuronidase activity showed that wounding caused local activation of the HRGP4.1 promoter in the phloem. Infection by tobacco mosaic virus was a less effective inducer than wounding. Stress induction was superimposed on tissue specific developmental expression in stem nodes and root tips, suggesting that HRGP4.1 may have specific structural roles in development as well as protective functions in defense. Deletion analysis showed that control of tissue specificity and wound inducibility lies in a region between -94 and -251 relative to the transcription start site and that activation by infection lies outside that region. PMID- 7480332 TI - Function of phytochrome A in potato plants as revealed through the study of transgenic plants. AB - We have generated transgenic potato plants (Solanum tuberosum) containing the potato phytochrome protein encoded by the PHYA gene cDNA (phyA) in sense or antisense orientation under the control of the 35S cauliflower mosaic virus promoter. Plants with increased and decreased phyA levels were analyzed. When grown under white light, development and growth of sprouts and plants were barely distinguishable from wild type. Under continuous far-red light, stem extension, leaf expansion, and hook opening of sprouts were accelerated in phyA overexpressors and delayed in antisense plants. Sprouts with reduced phyA levels were less sensitive to red light with regard to stem extension and expression of the small subunit genes for ribulose bisphosphate carboxylase. Under low red light:far-red light ratios, increased phyA levels reduced the stem extension component of the shade-avoidance response, whereas decreased levels led to an increase in the response. PMID- 7480333 TI - Transgenic analysis of a hybrid poplar wound-inducible promoter reveals developmental patterns of expression similar to that of storage protein genes. AB - The wound-inducible win3 multigene family from hybrid poplars (Populus trichocarpa x Populus deltoides) encodes proteins with structural similarities with Kunitz-type protease inhibitors (H.D. Bradshaw Jr., J.B. Hollick, T.J. Parsons, H.R.G. Clarke, M.P. Gordon [1990] Plant Mol Biol 14: 51-59), and at least one member, win3.12, is transcribed de novo in the injured and uninjured leaves of wounded trees (J.B. Hollick, M.P. Gordon [1993] Plant Mol Biol 22: 561 572). A previous study demonstrated that 1352 bp of 5' flanking DNA from the win3.12 gene confers local wound-regulated expression of the beta-glucuronidase gene in transgenic tobacco (Nicotiana tabacum cv Xanthi n.c.) (J.B. Hollick, M.P. Gordon [1993] Plant Mol Biol 22: 561-572). We extend this transgenic analysis here by examining the developmental regulation and systemic wound induction conferred by the same transgene construct in tobacco. Biochemical and histochemical surveys of beta-glucuronidase activity are described for four, independent transgenic lines. The observed spatial and temporal expression patterns coincide with dormant storage tissues and with previously described expression patterns for both seed and vegetative storage protein genes. Developmental northern blot analysis of win3 RNA levels in poplar seeds confirms that proper temporal expression of the reporter gene is maintained during tobacco seed maturation. These results demonstrate that a putative Kunitz-type protease inhibitor can be wound inducible in addition to being expressed in developing seeds. PMID- 7480334 TI - Plant antibodies for immunotherapy. AB - The original report of Hiatt (1989) initiated a wave of excitement at the realization that a complex mammalian protein such as immunoglobulin could be assembled within a plant cell. The general reaction was one of amazement, but interest in exploiting the possibilities arising from the discovery, for example to make antibodies of therapeutic value, has taken a considerable time to develop. In the meantime, other recombinant expression systems and traditional cell culture techniques have advanced and overcome some of their problems, particularly those associated with yields. Plants, however, still offer unique advantages, especially in their ability to match the protein assembly capabilities of mammalian cells (as demonstrated by the assembly of SIgA molecules), as well as to provide antibodies in bulk at low cost. In addition, the area of "immunization" of plants holds great promise and will surely be a field of enormous growth for the future. PMID- 7480335 TI - Lectins as plant defense proteins. PMID- 7480337 TI - Expression of an Arabidopsis potassium channel gene in guard cells. AB - The Arabidopsis thaliana KAT1 cDNA encodes a voltage-gated inward-rectifying K+ channel. A KAT1 genomic DNA clone was isolated and sequenced, and a 5' promoter and coding sequences containing eight introns were identified. Reporter gene analysis of transgenic plants containing the KAT1 promoter fused to bacterial beta-glucuronidase showed robust beta-glucuronidase activity primarily in guard cells. PMID- 7480336 TI - Characterization and gene expression of nodulin Npv30 from common bean. AB - We previously reported that transcripts for a 30-kD nodulin (Npv30) are very abundant in the nodule. In this paper we describe the isolation and characterization of Npv30 cDNA and genomic clones. Npv30 has the following characteristic features: (a) a putative signal sequence at the deduced amino terminal region, (b) a proline-rich stretch at the carboxy terminus, and (c) a characteristic domain of four cysteines that resemble metal-binding sites. In Phaseolus vulgaris L., Npv30 is encoded by a small gene family that shares discrete sequence homologies with another small gene family in soybean. An antibody against a beta-galactosidase-Npv30 fusion protein detected two proteins of 28 and 30 kD. Although Npv30 transcripts are very abundant, they encode proteins that are hardly detected in nodule fractions, suggesting that these proteins have a short half-life and/or the mRNAs are strongly regulated at the translational level. Npv30 transcripts were detected in the infected cells of the nodule by in situ hybridization experiments. PMID- 7480338 TI - Molecular and ultrastructural analysis of a nonchromosomal variegated mutant. Tomato mitochondrial mutants that cause abnormal leaf development. AB - Mutants were recovered in a population of cybrids formed following protoplast fusion between tomato (Lycopersicon esculentum Mill.) cv UC82 and Lycopersicon pennellii Corr. The cybrids were identified as individuals with recombinant cytoplasmic genomes but only tomato nuclear genomes. The mutants were identified based on two features, a variegated sectoring of light and dark green regions on their leaves, stems, and fruit, and reduced growth in the field. The mutants produced 50% of the shoot fresh weight and 20% of the fruit fresh weight of the parental type, UC82. The variegated sectoring was maternally inherited. The chloroplast genome in the mutants was indistinguishable from the chloroplast genome in UC82, when distribution of restriction endonuclease sites was used as an assay. The mitochondrial genome in the mutants, however, was recombinant, containing genes from UC82 and L. pennellii. Light microscopic analysis of the leaves of the mutants demonstrated an absence of the palisade layer in the variegated sectors. Electron microscopic analysis of these same regions demonstrated an absence of normal inner membranes in the mitochondria of these cells. PMID- 7480339 TI - Histones of Chlamydomonas reinhardtii. Synthesis, acetylation, and methylation. AB - Histones of the green alga Chlamydomonas reinhardtii were prepared by a new method and fractionated by reversed-phase high-performance liquid chromatography. Acid-urea-Triton gel analysis and tritiated acetate labeling demonstrated high levels of steady-state acetylation for the single histone H3 protein, in contrast to low levels on histones H4 and H2B. Twenty percent of histone H3 is subject to dynamic acetylation with, on average, three acetylated lysine residues per protein molecule. Histone synthesis in light-dark-synchronized cultures was biphasic with pattern differences between two histone H1 variants, between two H2A variants, and between H2B and ubiquitinated H2B. Automated protein sequence analysis of histone H3 demonstrated a site-specific pattern of steady-state acetylation between 7 and 17% at five of the six amino-terminal lysines and of monomethylation between 5 and 81% at five of the eight amino-terminal lysines in a pattern that may limit dynamic acetylation. An algal histone H3 sequence was confirmed by protein sequencing with a single threonine as residue 28 instead of the serine28-alanine29 sequence, present in all other known plant and animal H3 histones. PMID- 7480340 TI - Identification and gene expression of anaerobically induced enolase in Echinochloa phyllopogon and Echinochloa crus-pavonis. AB - Enolase (2-phospho-D-glycerate hydrolase, EC 4.2.1.11) has been identified as an anaerobic stress protein in Echinochloa oryzoides based on the homology of its internal amino acid sequence with those of enolases from other organisms, by immunological reactivity, and induction of catalytic activity during anaerobic stress. Enolase activity was induced 5-fold in anoxically treated seedlings of three flood-tolerant species (E. oryzoides, Echinochloa phyllopogon, and rice [Oryza sativa L.]) but not in the flood-intolerant species (Echinochloa crus pavonis). A 540-bp fragment of the enolase gene was amplified by polymerase chain reaction from cDNAs of E. phyllopogon and maize (Zea mays L.) and used to estimate the number of enolase genes and to study the expression of enolase transcripts in E. phyllopogon, E. crus-pavonis, and maize. Southern blot analysis indicated that only one enolase gene is present in either E. phyllopogon or E. crus-pavonis. Three patterns of enolase gene expression were observed in the three species studied. In E. phyllopogon, enolase induction at both the mRNA and enzyme activity levels was sustained at all times with a further induction after 48 h of anoxia. In contrast, enolase was induced in hypoxically treated maize root tips only at the mRNA level. In E. crus-pavonis, enolase mRNA and enzyme activity were induced during hypoxia, but activity was only transiently elevated. These results suggest that enolase expression in maize and E. crus-pavonis during anoxia are similarly regulated at the transcriptional level but differ in posttranslational regulation, whereas enolase is fully induced in E. phyllopogon during anaerobiosis. PMID- 7480341 TI - A potent antimicrobial protein from onion seeds showing sequence homology to plant lipid transfer proteins. AB - An antimicrobial protein of about 10 kD, called Ace-AMP1, was isolated from onion (Allium cepa L.) seeds. Based on the near-complete amino acid sequence of this protein, oligonucleotides were designed for polymerase chain reaction-based cloning of the corresponding cDNA. The mature protein is homologous to plant nonspecific lipid transfer proteins (nsLTPs), but it shares only 76% of the residues that are conserved among all known plant nsLTPs and is unusually rich in arginine. Ace-AMP1 inhibits all 12 tested plant pathogenic fungi at concentrations below 10 micrograms mL-1. Its antifungal activity is either not at all or is weakly affected by the presence of different cations at concentrations approximating physiological ionic strength conditions. Ace-AMP1 is also active on two Gram-positive bacteria but is apparently not toxic for Gram-negative bacteria and cultured human cells. In contrast to nsLTPs such as those isolated from radish or maize seeds, Ace-AMP1 was unable to transfer phospholipids from liposomes to mitochondria. On the other hand, lipid transfer proteins from wheat and maize seeds showed little or no antimicrobial activity, whereas the radish lipid transfer protein displayed antifungal activity only in media with low cation concentrations. The relevance of these findings with regard to the function of nsLTPs is discussed. PMID- 7480342 TI - Levels of a terpenoid glycoside (blumenin) and cell wall-bound phenolics in some cereal mycorrhizas. AB - Four cereals, Hordeum vulgare (barley), Triticum aestivum (wheat), Secale cereal (rye), and Avena sativa (oat), were grown in a defined nutritional medium with and without the arbuscular mycorrhizal fungus Glomus intraradices. Levels of soluble and cell wall-bound secondary metabolites in the roots of mycorrhizal and nonmycorrhizal plants were determined by high-performance liquid chromatography during the first 6 to 8 weeks of plant development. Whereas there was no difference in the levels of the cell wall-bound hydroxycinnamic acids, 4-coumaric and ferulic acids, there was a fungus-induced change of the soluble secondary root metabolites. The most obvious effect observed in all four cereals was the induced accumulation of a terpenoid glycoside. This compound was isolated and identified by spectroscopic methods (nuclear magnetic resonance, mass spectrometry) to be a cyclohexenone derivative, i.e. blumenol C 9-O-(2'-O-beta glucuronosyl)-beta-glucoside. The level of this compound was found to be directly correlated with the degree of root colonization. PMID- 7480343 TI - Galactinol synthase from kidney bean cotyledon and zucchini leaf. Purification and N-terminal sequences. AB - Galactinol synthase (GS) was purified 1591-fold with a 3.9% recovery from the cotyledon of kidney bean (Phaseolus vulgaris) by a novel scheme consisting of ammonium sulfate fractionation followed by diethylaminoethyl, Affi-Gel Blue, and UDP-hexanolamine affinity chromatography. The purified enzyme had a specific activity of 8.75 mumol mg-1 min-1, a pH optimum of 7.0, and requirements for manganese ion and DTT. The enzyme exhibited a Km = 0.4 mM for UDP-galactose and a Km = 4.5 mM for myo-inositol. It was identified as a 38-kD peptide that co purified with a 41- and a 43-kD peptide as shown by sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE). Purification to homogeneity was achieved by isolating the 38-kD peptide from the SDS-PAGE gel. To clarify conflicting reports in the literature about the relative molecular mass of purified GS from zucchini leaf (Cucurbita pepo), a similar scheme with modified eluting conditions was used to purify GS from this source. Zucchini leaf GS was purified to homogeneity and identified as a 36-kD peptide on SDS-PAGE. Partial N terminal sequences of the 38-kD peptide from kidney bean cotyledon and the 36-kD peptide from zucchini leaf were obtained. To facilitate identification of GS during the purification, an assay utilizing thin-layer chromatography and an isotopic analytic imaging scanner was developed. PMID- 7480344 TI - Polyphenol oxidase in potato. A multigene family that exhibits differential expression patterns. AB - Polyphenol oxidase (PPO) activity in potato (Solanum tuberosum) plants was high in stolons, tubers, roots, and flowers but low in leaves and stems. PPO activity per tuber continued to increase throughout tuber development but was highest on a fresh weight basis in developing tubers. PPO activity was greatest at the tuber exterior, including the skin and cortex tissue 1 to 2 mm beneath the skin. Flowers had high PPO activity throughout development, particularly in the anthers and ovary. Five distinct cDNA clones encoding PPO were isolated from developing tuber RNA. POT32 was the major form expressed in tubers and was found in all parts of the tuber and at all stages of tuber development. It was also expressed in roots but not in photosynthetic tissues. POT33 was expressed in tubers but mainly in the tissue near the skin. POT72 was detected in roots and at low levels in developing tubers. NOR333 was identical with the P2 PPO clone previously isolated from potato leaves (M.D. Hunt, N.T. Eannetta, Y. Haifeng, S.M. Newman, J.C. Steffens [1993] Plant Mol Biol 21: 59-68) and was detected in young leaves and in tissue near the tuber skin but was highly expressed in flowers. The results indicate that PPO is present as a small multigene family in potato and that each gene has a specific temporal and spatial pattern of expression. PMID- 7480345 TI - Intracellular carbonic anhydrase of Chlamydomonas reinhardtii. AB - An intracellular carbonic anhydrase (CA; EC 4.2.1.1) was purified to homogeneity from a mutant strain of Chlamydomonas reinhardtii (CW 92) lacking a cell wall. Intact cells were washed to remove periplasmic CA and were lysed and fractionated into soluble and membrane fractions by sedimentation. All of the CA activity sedimented with the membrane fraction and was dissociated by treatment with a buffer containing 200 mM KCI. Solubilized proteins were fractionated by ammonium sulfate precipitation, anionic exchange chromatography, and hydrophobic interaction chromatography. The resulting fraction had a specific activity of 1260 Wilbur-Anderson units/mg protein and was inhibited by acetazolamide (50% inhibition concentration, 12 nM). Final purification was accomplished by the specific absorption of the enzyme to a Centricon-10 microconcentrator filter. A single, 29.5-kD polypeptide was eluted from the filter with sodium dodecyl sulfate-polyacrylamide gel electrophoresis sample buffer, and a 1.5 M ammonium sulfate eluate contained CA activity. In comparison with human CA isoenzyme II, the N-terminal and internal amino acid sequences from the 29.5-kD polypeptide were 40% identical with the N-terminal region and 67% identical with an internal conserved region. Based on this evidence, we postulate that the 29.5-kD polypeptide is an internal CA in C. reinhardtii and that the enzyme is closely related to the alpha-type CAs observed in animal species. PMID- 7480346 TI - A putative vacuolar processing protease is regulated by ethylene and also during fruit ripening in Citrus fruit. AB - A putative citrus vacuolar processing thiolprotease cDNA (Cit-vac) was isolated from a cDNA library of Citrus fruits (Citrus sinensis L. Osbeck var Washington navel). The cDNA is 58 and 57% identical with vacuolar processing seed proteases from castor bean and soybean, respectively. The Citvac sequence shows a typical signal peptide for entering into the endoplasmic reticulum and two glycosylation signals. Using an in vitro transcription-translation system, we show that the Citvac precursor is able to enter a microsomal fraction and to undergo proteolytic processing and glycosylation. Transcript levels for the Citvac are developmentally regulated in the flavedo (outer colored part of the fruit peel) and increase during fruit ripening and in the flower during opening. Exogenous treatment with ethylene induces Citvac mRNA expression in both fruits and leaves. Citvac is encoded by one or two genes in the Citrus genome. The possible role of the Citvac gene product during fruit ripening and other ethylene-mediated processes is discussed. PMID- 7480347 TI - Coi1-dependent expression of an Arabidopsis vegetative storage protein in flowers and siliques and in response to coronatine or methyl jasmonate. AB - The phytotoxin coronatine and the plant growth regulator methyl jasmonate (MeJA) inhibit the growth of Arabidopsis seedlings. Coronatine and MeJA induced the accumulation of an approximately 29-kD protein in wild-type seedlings but not in seedlings of the coi1 mutant, which is insensitive to both compounds. The approximately 29-kD protein was recognized not only by antibodies raised against the partially purified polypeptide, but also by antibodies raised against vegetative storage proteins (VSPs) from soybean (29 kD) and poplar (32 kD). In the absence of added MeJA/coronatine, the VSP-like protein was highly expressed in flowers and siliques but not in seeds, seedlings, or mature leaves of wild type Arabidopsis. By contrast, this protein could not be detected in coi1 seedlings treated with coronatine or MeJA, and it was found in very low levels in the male sterile flowers of coi1. A transcript corresponding to the gene of the Arabidopsis 27-kD VSP precursor shows the same pattern of expression as the VSP like protein. Significantly, the VSP-like protein was not detected in green siliques or seeds obtained from coi1 flowers fertilized with wild-type pollen. We conclude that the VSP-like protein is normally expressed in maternal tissues, where it is regulated by COI1, but is not essential for the development of siliques. PMID- 7480348 TI - Targeting and release of phytohemagglutinin from the roots of bean seedlings. AB - Phytohemagglutinin (PHA), an abundant vacuolar seed protein of the common bean (Phaseolus vulgaris), is a tetramer of two homologous polypeptides, PHA-E and PHA L. The roots of bean seedlings release into the culture medium a cross-reacting lectin that is most closely related to PHA-E. Reverse-transcriptase polymerase chain reaction with root mRNA as template was used to identify PHA transcripts in the roots of bean seedlings. Roots were found to contain mRNA for PHA-E but not for PHA-L. Indirect immunocytochemical detection with colloidal gold and antibodies to deglycosylated PHA showed that in the meristem of the primary root, PHA accumulates in vacuoles. However, in elongated root cells PHA was found only in the cell walls, indicating targeting to an alternate location. These results are discussed in relation to the various mechanisms that may account for the release of a normally vacuolar protein by roots. PMID- 7480349 TI - A rapeseed cold-inducible transcript encodes a phosphoenolpyruvate carboxykinase. AB - We have isolated a clone corresponding to a new cold-regulated gene from a cDNA library made from rapeseed (Brassica napus cv Samourai) cold-acclimated etiolated seedlings. Sequence analysis and homology searches showed that this clone encodes a protein highly homologous to the ATP-dependent phosphoenolpyruvate carboxykinase (PEPCK; EC 4.1.1.49) from Saccharomyces cerevisiae, Trypanosoma, Rhizobium sp., and Escherichia coli; we refer to the B. napus clone as BnPEPCK. A potential ATP-binding site existing in all PEPCK proteins was also found in BnPEPCK. Although there was a basal expression of BnPEPCK in seedlings grown at control, room temperature, the steady-state level of the transcripts increased at 4 degrees C and decreased to normal levels when the seedlings were returned to control temperature (22 degrees C). Using antibodies made against a recombinant histidine-BnPEPCK fusion protein, we demonstrated that BnPEPCK protein level is correlated with the accumulation of the BnPEPCK transcript. PMID- 7480350 TI - Molecular cloning and characterization of the cDNA coding for the biotin containing subunit of the chloroplastic acetyl-coenzyme A carboxylase. AB - We report the molecular cloning and sequence of the cDNA coding for the biotin containing subunit of the chloroplastic acetylcoenzyme A (CoA) carboxylase (ACCase) of Arabidopsis thaliana (CAC1). The 3' end of the CAC1 sequence, coding for a peptide of 94 amino acids, which includes a putative biotinylation motif, was expressed in Escherichia coli as a glutathione-S-transferase (GST) fusion protein. The resulting GST-CAC1 fusion protein was biotinylated in vivo, indicating that CAC1 codes for a biotin-containing protein. Antibodies generated to the GST-CAC1 protein reacted solely with the 38-kD biotin-containing polypeptide of Arabidopsis. Furthermore, these antibodies inhibited ACCase activity in extracts from Arabidopsis leaves. The deduced amino acid sequence of CAC1 has an apparent N-terminal chloroplast-targeting transit peptide. The CAC1 protein is coded by a single Arabidopsis gene, and its mRNA accumulates to the highest levels in organs that are undergoing rapid growth. The amino acid sequence of the CAC1 protein is most similar to the biotin carboxyl-carrier protein component of eubacterial ACCases. These characterizations identify CAC1 as the biotin-containing subunit of the plastidic, heteromeric ACCase of Arabidopsis. The results support the ancient origin of the two structurally distinct ACCases of plants. PMID- 7480351 TI - Identification and characterization of cDNAs encoding ethylene biosynthetic enzymes from Pelargonium x hortorum cv Snow Mass leaves. AB - Two Pelargonium 1-aminocyclopropane-1-carboxylate (ACC) synthase cDNAs (GAC-1 and GAC-2) were identified and characterized. GAC-1 is 1934 bp long with a 1446-bp open reading frame encoding a 54.1-kD polypeptide. GAC-2 is a 1170-bp-long ACC synthase polymerase chain reaction fragment encoding 390 amino acids. Expression of GAC-1 and GAC-2 together with a previously identified ACC oxidase (GEFE-1) was examined in different Pelargonium plant parts, and leaves were subjected to osmotic stress (sorbitol), metal ion stress (CuCl2), auxin (2,4 dichlorophenoxyacetic acid [2,4-D]), and ethylene. GAC-1 expression was not detectable in any of the plant parts tested, whereas high levels of GAC-2 were expressed in the leaf bud, young leaf, young floret, fully open floret, and senescing floret. GAC-2 was expressed to a lesser degree in fully expanded leaves or roots and was undetectable in old leaves and floret buds. GEFE-1 was detectable at all leaf ages tested, in young and fully open florets, and in the roots; however, the highest degree of expression was in the senescing florets. GAC-1 was induced by sorbitol. Both GAC-1 and GAC-2 were only slightly affected by CuCl2 and induced indirectly by 2,4-D. GEFE-1 was highly induced by sorbitol, CuCl2, and 2,4-D. GAC-1, GAC-2, and GEFE-1 were unaffected by ethylene treatment. These results suggest that GAC-1 is only induced by stress and that GAC-2 may be developmentally regulated, whereas GEFE-1 is influenced by both stress and development. PMID- 7480352 TI - Pseudoreversion substitution at large-subunit residue 54 influences the CO2/O2 specificity of chloroplast ribulose-bisphosphate carboxylase/oxygenase. AB - Chlamydomonas reinhardtii mutant 31-4E lacks ribulose-1,5-bisphosphate carboxylase/oxygenase (Rubisco; EC 4.1.1.39) holoenzyme due to a mutation in the chloroplast rbcL gene. This mutation causes a glycine54-to-aspartate substitution within the N-terminal domain of the Rubisco large subunit. In the present study, photosynthesis-competent revertants were selected to determine whether other amino acid substitutions might complement the primary defect. Revertants were found to arise from only true reversion or either of two forms of pseudoreversion affecting residue 54. One pseudorevertant has a glycine54-to-alanine substitution that decreases the accumulation of holoenzyme, but the purified Rubisco has near normal kinetic properties. The other pseudorevertant has a glycine54-to-valine substitution that causes an even greater decrease in holoenzyme accumulation. Rubisco purified from this strain was found to have an 83% decrease in the Vmax of carboxylation and an 18% decrease in the CO2/O2 specificity factor. These results indicate that small increases in the size of amino acid side chains can influence Rubisco assembly or stability. Even though such changes occur far from the active site, they also play a significant role in determining Rubisco catalytic efficiency. PMID- 7480354 TI - The electronic Plant Gene Register. PMID- 7480355 TI - [Changes in sequence of time in schizophrenia--from 'severance' to 'collapse' in flux of time]. PMID- 7480353 TI - Multiple isoforms of Arabidopsis casein kinase I combine conserved catalytic domains with variable carboxyl-terminal extensions. AB - Three cDNA clones encoding isoforms of casein kinase I (CKI) were isolated from Arabidopsis thaliana. One full-length clone, designated CKI1, contained an open reading frame of 1371 bp encoding a protein of 51,949 D with an isoelectric point of 9.7. In addition to the highly conserved catalytic domain (of about 300 amino acids), the Arabidopsis CKI isoforms contain 150 to 180 amino acid carboxyl terminal extensions, which show among themselves a lower level of sequence conservation. These extensions do not show any sequence similarity to nonplant CKI isoforms, such as rat testis CKI delta, which is their closest isolated homolog, or to yeast CKI isoforms. Three additional isoforms of Arabidopsis CKI were found in the data bases of expressed sequence tags and/or were isolated serendipitously in nonspecific screening procedures by others. One of them also shows a carboxyl-terminal extension, but of only 80 amino acids. Casein kinase activity was detected in the soluble fraction of Escherichia coli strains expressing the CKI1 protein. This activity showed the crucial properties of CKI, including the ability to phosphorylate the D4 peptide, a specific substrate of CKI, and inhibition by N-(2-aminoethyl)-5-chloroisoquinoline-8-sulfonamide, a specific CKI inhibitor. Like several recombinant CKI isoforms from yeast, CKI1 was able to phosphorylate tyrosine-containing acidic polymers. PMID- 7480356 TI - [Lewis-x, a sugar chain structure, suppresses the epileptic seizure in El mouse]. AB - El mouse is a mutant strain with genetic epileptogenicity. Epileptic convulsion of El mouse has been shown to be related to metabolical and structural abnormalities in the hippocampal neuron. Gamma-globulin purified from the human serum has a marked anti-convulsive effect in El mouse. Human IgG, 80% of the gamma-globulin, has the reactivity indicating the presence of Lewis-x structure in the sugar chain. In the present study, the author found that 3' fucosyllactose, the terminal structure of Lewis-x, suppressed the tonic-clonic convulsion of El mouse. Lewis-x is a ligand of granular membrane protein-140 on the surfaces of the vascular endothelial cell and platelet. The vascular endothelial cell and platelet produce platelet-derived growth factor-BB, which prolongs the life-span of the hippocampal neuron and accelerates the formation of its axon and dendrite. The vascular endothelial cell and platelet in El mouse may be activated by 3'-fucosyllactose, and may secrete platelet-derived growth factor BB, which, in turn, may reduce the sensitivity to have epileptic convulsions in El mouse. Moreover, the author found that the fraction having Lewis-x reactivity obtained from the serum of El mouse had much reactivity of NeuAc alpha 2-6Gal. The sialic acid in Lewis-x structures of El mouse may reduce the ability of the Lewis-x structure to activate the endothelial cell and platelet. The findings in this study indicate that sugar chain structures and their metabolisms are closely related to the mechanism of genetic seizure susceptibility of El mouse. PMID- 7480357 TI - [Mental health activities for evacuees of the volcanic eruption of Mt. Unzen Fugen--crisis intervention of a psychiatrist]. AB - In a regional mental health activity for residents who have been evacuated for over three years on account of the volcanic eruption of Mt. Unzen-Fugen, a crisis intervention of the author took place in 76 people (21 males and 55 females) between June 1992 and November 1994. Most of them were over 41 in both sexes and half of female subjects was over 61. Using ICD-10, 57.9% of total were tentatively diagnosed as having depressive disorders (F32, F34.1, F41.2 and F43.2). Of the 76 cases, 49 people improved with counseling, 22 people received a prescription of psychotropics from general practitioners and 5 people required outpatient care or hospitalization at psychiatric or geriatric institutions. The outcome as of November 1994 was 60 remitted cases including 6 relapse cases, 6 worsened cases and 10 unchanged cases. The case history and the reality of crisis intervention was introduced on some cases and the evacuated people's peculiar psychology, that mental scars and grief after experiencing the disaster are unspoken and kept locked in their hearts, was discussed. On the basis of the above results and discussion, I considered the meaning of visiting the evacuees' shelters and emphasized the importance of the following basic strategies of crisis intervention to evacuated people: 1. to give them counseling so that they can accept their devastating experiences as part of their life, 2. to give support to the whole family, 3. to work in close cooperation with general doctors in cases requiring drug therapy. Furthermore, it was insisted that it is necessary to establish a disaster mental health system to give support to evacuees who need psychiatric help on a long time scale not just immediately after the disaster. Finally, I pointed out the undeveloped level of disaster mental health activities in Japan and referred to future problems. PMID- 7480358 TI - Ethanol and acetaldehyde metabolism in chinese with different aldehyde dehydrogenase-2 genotypes. AB - Alcohol dehydrogenase (ADH) and aldehyde dehydrogenase (ALDH) are the major enzymes responsible for ethanol metabolism in humans. Both enzymes exhibit genetic polymorphisms among racial populations. About half of the Chinese population lack mitochondrial ALDH2 activity and such a deficiency has been believed to be a negative risk factor for the development of alcoholism. To assess ethanol and acetaldehyde metabolism in Chinese with different ALDH2 genotypes, we genotyped 273 male adults at the ADH2, ADH3, and ALDH2 loci by using polymerase chain reaction-directed mutagenesis and restriction fragment length polymorphisms. Of the 143 individuals homozygous for both the ADH2*2 and the ADH3*1 alleles, 80, 55, and 8 were identified as ALDH2*1/*1, ALDH2*1/*2, and ALDH2*/*2, respectively. Five each from the above three ALDH2 genotypic subjects underwent alcohol elimination testing. Blood ethanol and acetaldehyde levels were determined at various times up to 130 min after intaking a low dose of ethanol (0.2 g/kg body weight) by using head-space gas chromatography and high performance liquid chromatography with fluorescence detection, respectively. The mutant homozygotes of ALDH2*2/*2 and the heterozygotes exhibited significantly higher peak acetaldehyde concentrations and also greater areas under the blood concentrations-time curve (AUC) than did the normal homozygotes of ALDH2*1/*1, with the mutant homozygotes both being the largest. The mutant homozygotes displayed significantly higher peak ethanol levels and AUC compared to the normal homozygotes. Of the 17 subjective feeling items tested, palpitation, facial warming, effects of alcohol, and dizziness were found to be most pronounced among the mutant homozygotes.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7480359 TI - A study on Huntington's disease associated trinucleotide repeat within the Chinese population. AB - Analysis of the polymorphic (CAG)n repeat in the huntingtin gene within the Chinese population in Taiwan confirmed the presence of an expanded repeat on all Huntington's disease (HD) chromosomes. Measurement of the specific CAG repeat sequence in 35 HD chromosomes from 11 unrelated families and 159 control chromosomes showed a range of from 9 to 29 (with a median of 17) repeats in normal subjects and 40 to 58 (with a median of 44) in affected subjects. The size distributions of normal and affected alleles did not overlap. The change in the size of the repeat in the HD size range on transmission for both sexes was variable. The expansion size inversely correlated with age at the onset in HD, especially from the early-onset disease. In summary, the molecular biology of HD is indistinguishable in Chinese and Caucasian populations, but the idea that world-wide HD evolved from a very limited number of European founders is no longer tenable. PMID- 7480360 TI - Effects of ascites and body position on gas exchange in patients with cirrhosis. AB - The effects of ascites and body position on gas exchange were evaluated in 22 male cirrhotics. All underwent spirometry and plethysmography in the sitting (erect) position. Single breath carbon monoxide diffusing capacity (DLco) was measured in the erect and supine positions in a random sequence. A significantly negative correlation was found between Kco (DLco corrected by alveolar volume [VA]) and lung volumes. DLco was significantly higher in the erect than in the supine position. On the other hand, Kco was significantly higher in the supine position. The differences of DLco and Kco between the erect and supine positions (the data in the supine minus those in the erect position) correlated highly with FEV1 and FVC. Thirteen patients underwent large volume paracentesis, and pulmonary function tests were done one day before and after paracentesis. Lung volumes and DLco increased significantly after paracentesis. In contrast, Kco decreased significantly. A significantly negative correlation was found between the change of Kco before and after paracentesis and that of lung volumes (the data after minus those before paracentesis). There was no remarkable change in the differences of DLco and Kco between the two positions after paracentesis. It is concluded that ascites may prevent further worsening of gas exchange and attenuate the postural effect on gas exchange in cirrhotic patients with significant amounts of ascites. PMID- 7480361 TI - Evaluation with receptor binding assay on the water extracts of ten CNS-active Chinese herbal drugs. AB - In the present study, we established receptor binding assays to evaluate the water extracts of ten central nervous system (CNS)-active Chinese herbal drugs. These ten herbal drugs are Chaihu (Radix Bupleuri), Chuanxiong (Rhizoma Chuanxiong), Danggui (Radix Angelicae sinensis), Danshen (Radix Salviae miltiorrhizae), Duhuo (Radix Angelicae pubescentis), Hangqin (Radix Scutellariae), Qinjiao (Radix Gentianae macrophyllae), Shengma (Rhizoma Cimicifugae), Suanzaoren (Semen Zizphi spinose), and Yangjihua (Flos Daturae). The results indicated that these water extracts contained the principles acting on the dopamine (D1 & D2), muscarinice acetylcholine (M1), or 5-HT (5-HT1A & 5 HT2) receptors, or the benzodiazepine and the gamma-amino-n-butyric acid (GABA) binding sites of GABAA receptors as determined by receptor binding assays. The receptors or binding sites which predominantly acted by each water extract are listed as follows: Chaihu: D2, 5-HT1A, GABA; Chuanxiong: GABA, 5-HT1A; Danggui: GABA, 5-HT1A; Danshen: BDZ; Duhuo: GABA, 5-HT1A, D2, D1; Hangqin: BDZ, D1, 5 HT1A; Qinjiao: GABA, BDZ, 5-HT1A, D2; Shengma: 5-HT1A; Suanzaoren: 5-HT1A, 5-HT2, GABA; Yangjihua: M1, 5-HT1A, 5-HT2. These results provided evidence to explain the CNS effects of these herbal drugs at the receptor level. Furthermore, these results provided information to direct the isolation and purification of receptor interactive compounds from these herbal drugs. PMID- 7480362 TI - Antihypertensive effects of AT-112, a newly synthesized quinazoline derivative, in spontaneously hypertensive rats. AB - The purpose of this study was to investigate the anti-hypertensive effect and possible mechanism of action of 2-(4-phenyl-1-piperazinyl)methyl-2,3 dihydroimidazo[1,2- c]quinazolin-5(6H)-one (AT-112), a newly synthesized ketanserin derivative, on in vivo and in vitro models. In vivo, central and peripheral administration of AT-112 produced a dose-dependent decrease of arterial blood pressure in conscious spontaneously hypertensive rats (SHR). In in vitro study, the vasodilating effect of AT-112 on isolated thoracic aorta was endothelium-independent. AT-112 was found to be a potent alpha 1-adrenoceptor blocking agent in the rat thoracic aorta as revealed by its competitive antagonism of phenylephrine (pA2 = 9.82 +/- 0.19), but was found to have noncompetitive antagonism to 5-HT. The IC50 of ketanserin and AT-112 to phenylephrine were 27.8 nM and 0.36 nM, respectively, and to 5-HT were 5.73 nM and 0.44 microM, respectively. Competition binding studies demonstrated that the affinity of AT-112 to alpha 1-adrenoceptor (Ki = 2.01 +/- 0.09 nM) was significantly higher than that of alpha 2-adrenoceptor (Ki = 3.86 +/- 0.64 MM). Based on the results of these in vivo, in vitro and competition binding studies, AT-112 is a potent antihypertensive agent, and its antihypertensive action is mainly mediated by the blockade of the alpha 1-adrenoceptor. PMID- 7480363 TI - Production, purification and characterization of two proteinaceous hen-egg-white lysozyme inhibitors from Pseudomonas aeruginosa M-1001. AB - Two proteinaceous lysozyme inhibitors, hen-egg-white lysozyme inhibitors F-I and F-II, were isolated from the culture broth of a bacterial strain identified as Pseudomonas aeruginosa M-1001. Maximum lysozyme inhibitory activity was obtained when the bacterium was grown aerobically in a medium consisting of 0.25% glucose, 0.25% beef extract, 0.25% polypepton, 1.0% sodium L-glutamate, and 1.0% soluble starch (pH 7.0) at 37 degrees C after 20-24 hrs. F-I and F-II were purified 20 and 7.5-fold, respectively, from the culture supernatant of P. aeruginosa M-1001 by ammonium sulfate fractionation, DEAE-Sepharose CL-6B column chromatography, and Sephacryl S-200 gel chromatography. The molecular weights of F-I and F-II were estimated to be about 57,000 and 33,000, by SDS-PAGE, respectively. F-I was stable in a pH range between 6 and 10 and below 50 degrees C. F-II was stable in a pH range between 6 and 11 and below 40 degrees C. Many Gram-positive bacteria were found to be inhibited by the crude lysozyme inhibitors. PMID- 7480364 TI - Expression of matrix proteins in cloned fibroblasts derived from periodontal tissue under different cell growth densities. AB - Extracellular matrix (ECM) proteins have been implicated in the attachment, migration and differentiation of cells during periodontal wound healing. This study was designed to investigate the expression of ECM proteins in gingival fibroblasts (GF) and periodontal ligament fibroblasts (PF) at different cell growth densities. Both mass-cultured and cloned cells were derived from explants of healthy human gingivae and periodontal ligament. The grown cells, in situ, were subjected to immunofluorescent staining for the expression of collagen type I (CI), III (CIII) and fibronectin (Fn), and then measured by microfluorimetry. Partial biochemical measurement of the ECM proteins secreted in the medium was done by SDS-PAGE and densitometry. These data indicated that CI, CIII and Fn were expressed in both mass-cultured and cloned GF and PF. There were variations in the expression of these ECM proteins among the clones. PF and most of its clones produced a greater amount of total cellular and ECM proteins than did GF. The expression of these proteins was found to be greater in areas of low, as opposed to high, cell density. Parallel results were also noted in limited biochemical analyses. Thus, PF differed from GF with regard to the production of cellular and matrix proteins, and protein metabolism was affected by cell growth density. These data tend to support the previous hypothesis that PF is the essential cell type contributing to periodontal regeneration. PMID- 7480366 TI - [Drug dependence in psychiatric practice--new trends in therapy of dependence]. PMID- 7480365 TI - The effect of cobalt-60 irradiation on bone marrow cellularity and alveolar osteoclasts. AB - The purpose of this study was to investigate the mechanisms of bone resorption in the rat dentoalveolar complex occurring as a result of orthodontic appliance therapy. Utilizing whole body radioactive cobalt 60 (Co60) irradiation in combination with orthodontic tooth movement, the activation, recruitment, and life span of osteoclasts was studied. Thirty-four adult Fischer 344 rats were irradiated with 10 and 20 Gray of Co60. Twelve days after irradiation, each rat was fitted with an orthodontic appliance; the rats were sacrificed 7, 14, or 21 days after appliance placement. To serve as controls, another group of 12 rats was subjected to orthodontic treatment only. Histologic sections were prepared from decalcified maxillary alveolar process, and osteoclasts were counted. In the control group, osteoclasts were presence in the periodontal membrane for four weeks after appliance placement, and the largest number of osteoclasts was observed in the second week. The smaller total irradiation dose (10 Gray, administered in 5 daily fractions) induced a transient reduction in the bone marrow cell count of more than 90%, followed by a complete rebound. The peak osteoclst number also was increased 110%. In contrast, the higher total dose (20 Gray, administered in 10 daily fractions) resulted in a reduction of 60% in bone marrow cellularity and a decrease in the peak osteoclast number by 40%. Fluctuations in bone marrow cellularity generally corresponded to similar variations in the osteoclast number, but there was an apparent lack of correspondence between bone marrow cell and white blood cell values. The total period of osteoclastic presence in the periodontal membrane following orthodontic activation, normally four weeks in duration, was reduced by one week in the irradiated animals. These findings lead us to speculate that the mechanisms of osteoclast activation and recruitment following orthodontic appliance therapy may involve three consecutive waves of osteoclast maturation. The total duration of osteoclastic bone resorption lasts 4 weeks, but the osteoclast lie span is calculated to be 9 to 10 days (9 days x 3 waves = 27 days). If the results of this study are proved to be reproducible, a scientific basis may have been provided to support the common practice in orthodontics wherein the duration between orthodontic appointments is 4-5 weeks. PMID- 7480367 TI - [Drug substitution treatment of heroin dependent patients with codeine preparations--treatment effects from the viewpoint of the physicians and patients]. AB - The controversy caused by the debate on treatment and substitution strategies in drug addiction is unchanged. This is particularly true concerning the use of codein-based drugs in the substitution of heroin abusers. However, only a few studies on the effects of this old and widely used method of substitution have been carried out. This article presents one retrospective and prospective study on the effects of the codein-based substitution in heroin abusers (n = 416). With respect to the issues addressed by this study such as somatic and psychic health, social integration, delinquincy and consumption patterns, patients as well as clinicians report an improvement in general health and fewer of the problems usually associated with heroin abuse, similar to the results from substitution treatment elsewhere. PMID- 7480368 TI - [Motivational work with alcohol dependent patients]. AB - Motivational work is secondary preventive work. The aim is to increase activity for maintaining long-term abstinence. The principles and structure of a secondary preventive program with several opportunities of intervention in the frame of detoxification treatment are presented. PMID- 7480369 TI - [Results of integrated inpatient detoxification and motivation treatment of alcohol dependent patients]. AB - During the course of the disease many alcoholic patients are detoxified on an inpatient basis. Detoxification lasts for some days to 2 or 3 weeks. The treatment goal of abstinence usually cannot be achieved by detoxification alone. Therefore a qualified detoxification should consist of a medical treatment, a careful (psychiatric) diagnostic process and a psychotherapeutic treatment aiming at motivation for further treatment. The described program integrates these components. The psychotherapeutic approach is emphatic and directive, trying to convince all patients to apply for further alcoholism treatment. Follow-up data of 120 of the initial 130 patients eight months after detoxification showed, that 68 patients (57%) started further treatment, most of them as inpatients. In the group of prognostic unfavourable patients, who had not received any disease related counseling before detoxification (77% of the sample), 47% started further alcoholism treatment. PMID- 7480370 TI - [Treatment of drug dependent patients in a psychiatric department]. AB - The authors report their three years experience with treating addicts (both alcoholics and users of prescription and illegal drugs) in a specialized unit employing a low-threshold admission policy. Difficulties as well as advantages of the concept are discussed. For the common treatment of patients addicted to different groups of substances on the same ward to be successful certain prerequisites probably have to be met: A clearly structured daily routine respectively therapy-program and close professional cooperation within a multifunctional therapeutic team harbouring a basic attitude of acceptance towards their patients. PMID- 7480371 TI - [Forensic psychiatric expert evaluation of so-called drug-induced psychoses]. AB - A considerable number of schizophrenic persons also abuse illegal drugs. This comorbidity raises problems of diagnosis which become particularly important when concerning forensic questions. The hypothesis of a distinct psychopathological syndrome of drug induced psychosis is widely criticized in the literature. A case report is presented to demonstrate the problems of differential diagnoses and the difficulties for psychiatric expertizing arising from these problems. PMID- 7480372 TI - [Management dialogue within multi-professional teams: contribution of psychoanalysis to integrated therapy of psychotic patients]. AB - By means of case studies and experiences in supervising multiprofessional teams in psychiatric wards it is pointed out that the institutional frame of psychiatry is dynamically connected with the intrapsychic conflicts of the psychotic patients and their interpersonal defence. The understanding of the teammembers' countertransference prevents collusive patterns of relationship. This symbolizing process which is permanently endangered by the psychotic splitting mechanisms of the patients as well as by the institutional defence contributes to develop a "common language" in the multiprofessional teams. Furthermore psychoanalytic supervision enables the multiprofessional staff to understand the intrapsychic and the interactional dimension of the behaviour of the psychotic patient and to maintain their empathy even under difficult conditions. PMID- 7480373 TI - [Social adjustment of psychiatric patients: evaluation of a modified version of the GAF (Global Assessment of Functioning) Scale]. AB - A modified version of the Global Assessment of Functioning Scale (GAFS-M) which is proposed as axis V for DSM-IV by Goldman et al. (1992), was analyzed in a consecutive series of 266 psychiatric patients. Scores were clearly dependent on ICD-10 diagnosis and on other psychiatric variables. The scale was almost independent on self-rating in relevant psychopathological dimensions. Altogether, the results favour the view that the modified version of the GAFS is an economic and sufficient instrument for assessing adaptive functioning in psychiatric patients. PMID- 7480374 TI - [Initial experiences with an electronic information system in a psychiatric treatment clinic]. AB - Inspite of the inherent potential, the computer-based medical record is underutilized today, although the use of computers in medicine and psychiatry is widespread. Lack of information is one of the reasons why physicians have resisted the introduction of information systems in medical practice beyond auxiliary services. The author describes the functions of a clinical information system currently in use in a psychiatric inpatient hospital, offering an electronic medical record, office automation tools, knowledge databases and individual PC-programs. He discusses measures to preserve the confidentiality and integrity of the electronic data and shortterm experiences concerning benefits and risks of the system. PMID- 7480376 TI - [From explanation to mythology--sexual abuse two more times and a helpful manual]. PMID- 7480375 TI - ["Masked" or "hidden" schizophrenia: initial diagnosis of a 12 year psychoses based on negative symptoms]. AB - We report th case of a 45-year-old woman who was referred to our hospital for a treatment of analgetic substance abuse. Surprisingly she reported after some time that she had been hearing imperative and commenting voices for 12 years. We discuss the importance of negative symptoms for the diagnosis of schizophrenia. PMID- 7480377 TI - Ketone reductase activity and reduced haloperidol/haloperidol ratios in haloperidol-treated schizophrenic patients. AB - Twenty schizophrenic patients were treated with a fixed haloperidol (HL) dose of 20 mg/day for 4 weeks. The conversion of HL to its reduced metabolite (reduced haloperidol, RH) occurs via the ketone reductase enzyme. RH is also converted back to HL by the cytochrome P450 2D6 isozyme. Ketone reductase activity can be measured in red blood cells. Plasma HL and RH levels were assayed by high performance liquid chromatography. Blood samples were obtained at baseline and during weeks 2 and 4 of HL therapy. Seventeen of 20 patients had ketone reductase values < 3. A significant correlation between ketone reductase and RH/HL plasma ratios was observed at week 4 in these 17 patients. Patients with ketone reductase activity < 3 could represent a subgroup of patients that metabolize HL differently. The wide interpatient variability observed with HL and RH plasma levels in HL-treated patients could reflect differences in ketone reductase activity and the metabolic status of debrisoquin hydroxylase (cytochrome P450IID6) in psychiatric patients. PMID- 7480378 TI - Lateral ventricular enlargement in schizotypal personality disorder. AB - Although an increase in the ratio of ventricular space to brain (ventricle-brain ratio), VBR) on computed tomography (CT) has been among the most robust findings in chronic schizophrenia, VBR has not been investigated in a large, well characterized clinical population of patients with schizotypal personality disorder (SPD), a clinical entity with a phenomenologic, gentle biological, and treatment response relationship to chronic schizophrenia. Accordingly, CT scans were obtained in 36 male SPD patients, 23 males with other personality disorders, 133 male schizophrenic patients, and 42 male normal volunteers. The mean body of the lateral VBR was significantly greater in the SPD patients than in the patients with other personality disorders. The VBR of the SPD patients did not differ significantly from either that of the normal volunteers or the schizophrenic patients but was intermediate between the two groups. There were no correlations with either psychotic-like or deficit-related symptoms of SPD in either the SPD or total personality disorder cohorts. SPD patients, like schizophrenic patients, may have increased VBRs compared wit patients with other personality disorders; their VBRs fall between the means of schizophrenic patients and normal control subjects. PMID- 7480379 TI - Discriminant analysis of MRI measures as a method to determine the presence of dementia of the Alzheimer type. AB - Multivariate discriminant analysis of brain volumes obtained from semiautomated magnetic resonance image (MRI) quantification was used in an attempt to identify demented patients very early in the course of the disease. Temporal and posterior frontal brain volumes were quantified from MRIs in a cross-sectional study of 31 male and female patients with dementia of the Alzheimer type (DAT) and 29 age- and sex-matched healthy comparison subjects. Mean scores on the Folstein Mini Mental State Examination (MMS) were in the mild range for the DAT group (20 +/- 6.6), but patients with moderate and severe dementia were also included (MMS range of entire DAT group = 4-28). Significant mean differences in frontal and temporal lobe brain volumes were found between the DAT group and the age- and sex matched healthy comparison group, but the sensitivity of any single measure was limited to 87% with a specificity of 83%. Initial multivariate discriminant analysis revealed significant gender differences among the healthy subjects, but not the DAT patients. The large group size allowed for subsequent discriminant analyses to be performed by gender. All healthy subjects and DAT patients were correctly classified by gender-specific discriminant functions. The male discriminant variables included brain volume, age, and temporal lobe measures. Inclusion of age in the male discriminant function accounted for age-related brain atrophy, a finding that may have emerged as a consequence of the broad age range of the male DAT population (50-81 years). The male discriminant function was also successfully applied to an independent group of mildly demented subjects that included patients for whom the diagnosis of dementia was uncertain but verified by follow-up clinical evaluations. Measures of temporal lobe brain matter and temporal lobe cerebrospinal fluid volumes were the significant discriminator variables for the women. Quantitative MRI and multivariate discriminant analysis showed promise in distinguishing the dementing process from healthy aging in a group of 60 subjects. Moreover, while not diagnostic of DAT, the approach appeared to offer additional information about the probability of a diagnosis being later confirmed in patients with very mild dementia for whom the clinical identification of DAT is uncertain. PMID- 7480380 TI - Distractibility in schizophrenia. AB - This study examined the effects of neuroleptic medication on the allocation of attentional resources to distracting stimuli in patients with schizophrenia. Twenty-five patients were tested twice (medication-free and after medication stabilization) on the Identical Pairs versions of the Continuous Performance Test under both distraction and no-distraction conditions. Sixteen patients were chronically ill adults and nine patients were young neuroleptic-native patients in the early stages of illness. Results indicated that neuroleptic treatment did not improve distractibility for either group and that both groups were comparably distractible. These findings suggest that medication does not improve the misallocation of attention to distracting stimuli in patients with schizophrenia. PMID- 7480381 TI - Memory and social skill in schizophrenia: the role of gender. AB - The relationship between social skill and memory was examined in a group of patients with schizophrenia and schizoaffective disorder. There were few or no differences between male and female patients in history of the illness, symptomatology, memory, and other cognitive factors, or social skill. Despite the absence of differences, impaired memory was related to poorer social skills for the women, but not the men. Reanalysis of data on social skill and memory from an earlier study (Mueser et al., 1991) revealed a similar pattern of results. The findings suggest that the contribution of cognitive factors to impaired social competence in schizophrenia may differ in men and women. Alternatively, the results may reflect sampling bias, whereby more severely ill women are studied than men, relative to their respective populations. PMID- 7480382 TI - Schizophrenic patients' perceptions of their relatives' attitudes. AB - Although the attitudes of family members as revealed by measures of expressed emotion (EE) have been shown to be associated with the course of schizophrenic illness, little is known about how the patients perceive these attitudes. A detailed interview was used to assess patients' perceptions of their family members' behaviors toward them. Family member's EE was assessed with the Five Minute Speech Sample (FMSS). Overall, patients' perceptions of criticism were congruent with a component of the FMSS-EE that measures criticism. Results indicated that when family members had high EE/critical scores, patients perceived them as displaying more instances of critical behavior. All cases in which patients' perceptions of criticism were incongruent with measures of FMSS EE occurred among ethnic minority group members. Although the FMSS-EE did not predict outcome in this study, patients who perceived their relatives as higher in criticism had more negative outcomes at 1 year. These findings suggest that when family environments are examined in patients from ethnic minority groups, the patients' perspective may be a more potent predictor of outcome than traditional measures of EE. PMID- 7480383 TI - Family psychiatric screening instruments for epidemiologic studies: pilot testing and validation. AB - Family history, a risk factor for psychiatric disorders, is infrequently assessed in epidemiologic studies due to time and cost constraints. We designed a brief computer-scorable instrument, the Family History Screen for Epidemiologic Studies (FHE), which collects a pedigree and screens for 15 DSM-III diagnoses in an informant and in his family members. The FHE was administered to one informant in 77 families in which we had collected pedigrees, interviewed 77 informants and 239 relatives using the Lifetime Anxiety version of the schedule for Affective Disorders and Schizophrenia or the Epidemiologic version of the Schedule for Affective Disorders and Schizophrenia for School-Aged Children, and performed best-estimate diagnoses. We evaluated the accuracy with which the FHE predicted best-estimate diagnoses. For adults reporting on themselves, the FHE demonstrated high levels of sensitivity and specificity for depression (67.4, 75.0) and panic (92.5, 89.2), and low sensitivity and high specificity for substance abuse (33.3, 93.6). For informants reporting on adult relatives, sensitivity was low and specificity was high for depression (35.2, 84.9), panic (20.0, 91.7), and substance abuse (42.1, 93.4). For informants reporting on children, perhaps due to lower prevalence, sensitivity and specificity were poor. The FHE is a good screen for psychiatric disorders in adult informants, but it is not useful for family history. It may be useful in primary care medical settings as a screen for psychiatric history. PMID- 7480384 TI - Dorsal periaqueductal gray-induced aversion as a simulation of panic anxiety: elements of face and predictive validity. AB - Neurosurgical stimulation of the dorsal periaqueductal gray (dPAG) matter in man induces acute signs of autonomic arousal and feelings of subjective anxiety; those signs have phenomenological similarity with the symptom profile characterizing a panic attack. Animals undergoing dPAG stimulation show comparable physical signs of autonomic activation and sudden fear-suggestive behavioral reactions that can be shaped into operant self-interruption behavior. Drugs known to acutely reduce (alprazolam, clonazepam) or precipitate (yohimbine, caffeine) panic attacks in patients were found to acutely and dose-dependently reduce or enhance, respectively, aversion induced by dPAG stimulation in rats. When considered as an animal model of panic anxiety, the dPAG model simultaneously meets criteria of face validity (symptomatic homology) and predictive validity (pharmacological homology under short-term treatment); aspects of its construct validity (theoretical rationale supporting the model) are discussed. It is suggested that dPAG stimulation-induced aversion may represent a model of some aspects of panic disorders. PMID- 7480385 TI - Effect of melatonin on active transport of serotonin into blood platelets. AB - Previous research has demonstrated that the circadian rhythm of melatonin negatively correlates with that of the sensitivity of platelet serotonin uptake sites. Moreover, serotonin has been found to be a competitive inhibitor of melatonin binding to platelets. The goal of the present study was to investigate whether melatonin directly affects platelet serotonin transport. Blood samples from 12 healthy subjects were assayed for both serotonin transport into platelets. Active transport of serotonin (at a concentration of 10(-6) M) into blood platelets was measured in the presence of melatonin at physiological concentrations (10(-12) M to 2 x 10(-9) M) and high nonphysiological concentrations (2 x 10(-8) M to 2 x 10(-3) M). Melatonin inhibited serotonin uptake by blood platelets at the high concentrations only ( > 10(-5) M with IC50 value of 1.1 x 10(-3) M) and had no effect at physiological concentrations. PMID- 7480386 TI - Biological correlates of slow wave sleep deficits in functional psychoses: 31P magnetic resonance spectroscopy. AB - Deficits in slow wave sleep (SWS) are consistently seen in schizophrenia and related psychotic disorders. However, the pathophysiological significance of this finding is uncertain. In 19 patients with psychotic illness, sleep and 31P magnetic resonance spectroscopy (MRS) studies were carried out before the patients began medication treatment. Polysomnographic studies were carried out in 2-3 consecutive nights. MRS studies were performed with a surface coil and a depth-resolved pulse sequence focusing on the dorsal prefrontal cortex. Phosphomonoesters were correlated with visually scored delta and Stage 4 sleep, as well as with automated delta wave counts. An inverse relation was also seen between negative symptoms scores and SWS. The association between decreases brain anabolic processes (reflected by decreased PME) and decreased SWS may be related either to processes of accelerated aging or to developmentally mediated alterations in cortical synaptic pruning, postulated to underlie the pathophysiology of functional psychoses. PMID- 7480387 TI - Brain morphology assessed by computed tomography in patients with geriatric depression, patients with degenerative dementia, and normal control subjects. AB - To determine whether patients with geriatric depression have specific alterations in brain morphology, internal (ventricles) and external (frontal, temporal, and parieto-occipital) components of the cerebrospinal fluid (CSF) spaces were examined. Planimetric measurements of computed tomographic (CT) scans from patients with geriatric depression were compared with measurements from two age- and sex-matched control groups: normal control subjects and patients with primary degenerative dementia. Scans of 34 patients (6 men, 28 women; mean age = 70.7 years) who met DSM-III-R diagnostic criteria for major depression, 29 patients with DSM-III-R primary degenerative dementia (8 men, 21 women; mean age = 71.2 years), and 43 nonpsychiatric control subjects (10 men, 33 women; mean age = 70.8 years) were evaluated. The areas of the frontal and parieto-occipital sulci, the Sylvian fissures, and the lateral and third ventricles were measured separately for the right and left hemispheres. Compared with the control subjects, patients with geriatric depression revealed a remarkable enlargement (up to 125%) of the left Sylvian fissure on several levels and a more subtle enlargement of the ventricles, cortical sulci, and right Sylvian fissure (20-50%). The laterality index differed significantly between depressed patients and normal control subjects (but not between the demented patients and the normal control group) only for the Sylvian fissure. Demented patients showed a considerable brain atrophy that affected all CSF components (enlargement of 30-160%) but the left temporal region was less affected than in the depressed patients. Compared with the findings in geriatric depression, ventricular enlargement was significant in dementia.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7480388 TI - Reliability of cerebral measures in repeated examinations with magnetic resonance imaging. AB - This study examined the reliability of quantitative measures of cerebral magnetic resonance images (MRI) in repeated scans. Ten subjects were scanned twice, at 2- to 4-week intervals. Volumetric data from 14 regions of the cerebrum, the caudate nucleus, and the lateral ventricles and area measures of the corpus callosum were acquired. Intrarater and scan-rescan reliabilities, including the relative percent error from each of these two sources, were determined for each structure. Intraclass correlations ranged from 0.88 for the head of the caudate nucleus to 0.99 for the ventricular volume. Quantitative cerebral MRI measures of these structures are stable over time intervals of 2-4 weeks. PMID- 7480389 TI - Against the laterality index as a measure of cerebral asymmetry. AB - The laterality index (right - left)/(right + left) has been widely used to measure cerebral asymmetry. However, its validity rests on the assumption that confounding effects of overall head and body size are symmetrical on both sides of the brain. We have found that this assumption may not always be justified. In magnetic resonance imaging data from a group of 43 right-handed normal control and schizophrenic subjects, the laterality index controlled poorly for the asymmetrical confounding effect of height, leading to a spurious (although statistically significant) correlation between height and apparent cerebral asymmetry. Regression modeling is advocated as a more rational and effective way to explore and control size effects confounding cerebral lateralization. PMID- 7480390 TI - No elevated D2 dopamine receptors in neuroleptic-naive schizophrenic patients revealed by positron emission tomography and [11C]N-methylspiperone. AB - The dopamine hypothesis of schizophrenia received strong support when a two- to three-fold elevation of D2 receptor densities was demonstrated by positron emission tomography (PET) and [11C]N-methylspiperone ([11C]NMSP). In the present study, the reproducibility of this finding was examined by application of a similar method in seven normal comparison subjects and seven neuroleptic-naive schizophrenic patients examined by PET before and after administration of haloperidol, 7.5 mg. After haloperidol, the specific binding of [11C]NMSP was reduced by 80-90%, resulting in a signal-to-noise ratio that was unfavorably low for reliable quantification. No significant difference was found between normal subjects and patients in a descriptive analysis of the time-activity curves or in a nonequilibrium graphical determination of D2 receptor densities in the basal ganglia. The results are consistent with those of a previous quantitative PET study of [11C]raclopride binding, which showed normal densities of D2 receptors in the striatum of neuroleptic-naive schizophrenic patients. PMID- 7480391 TI - Regional differentiation of cortical activity in schizophrenia: a complementary approach to conventional analysis of regional cerebral blood flow. AB - This study examines an alternative to a narrow locus-of-dysfunction cortical pathophysiology in schizophrenia, which in turn suggests a modified approach to the analysis of data from regional cerebral blood flow studies. The results provide qualified support for a model of impaired differentiation of cortical activity in schizophrenia. As an index of the differentiation of cortical fields, regional gradients of cortical blood flow were examined. Medicated patients (n = 15) failed to show the differentiation of networks in frontal areas during a verbal memory recognition task observed in normal comparison subjects (n = 15). Unmedicated patients (n = 15) at rest tended to lose normal lateralization and had increased gradients in the right frontal area compared with normal comparison subjects. PMID- 7480392 TI - Basal ganglia T2 relaxation times in schizophrenia: a quantitative magnetic resonance imaging study in relation to tardive dyskinesia. AB - Transverse, or spin-spin, relaxation times (T2) from magnetic resonance images of basal ganglia structures were compared between control subjects and patients with schizophrenia, who were subdivided on the basis of the presence or absence of tardive dyskinesia. As a group, schizophrenic patients showed evidence of somewhat more prolonged T2 relaxation times in the right putamen and globus pallidus than did control subjects; there were no significant correlations between hemispheric T2 values and corresponding volumes of the lateral ventricles. Overall, there was little difference in T2 values between patients with and without tardive dyskinesia. These data extend the range of evidence for basal ganglia dysfunction in schizophrenia, but they do not support earlier reports of prominent T2 changes associated with tardive dyskinesia. PMID- 7480393 TI - Roots of unitive experience. PMID- 7480394 TI - Stones in a stream. PMID- 7480395 TI - Thoughts without a thinker: Buddhism and psychoanalysis. PMID- 7480396 TI - In search of the self: Zen Buddhism and psychoanalysis. PMID- 7480397 TI - Repression or suppression? Freud's interpretation of the dream of Irma's injection. PMID- 7480398 TI - Blow up: the "I" of the camera. PMID- 7480399 TI - A follow-up of child analysis. The analyst as a real person. AB - Thirteen cases of child analysis or intensive psychotherapy are described briefly, along with follow-up data. Seven made spontaneous contact with the analyst, some as long as thirty or forty years after the termination of treatment; three more were followed into young adulthood through reports of family, therapists, or friends; and three more made no contact. The process of internalization and preservation of the analyst as a real person is discussed, as well as why it occurs in some cases and not others. Most of the late contacts were made to obtain help in decisions regarding further treatment for self or family members. PMID- 7480400 TI - Trauma in the preverbal period. Symptoms, memories, and developmental impact. AB - In this paper, five children who experienced traumas during the preverbal period are described. The clinical presentations are oriented around three questions: (1) What are the expectable symptomatic reactions of preverbal infants to trauma? (2) To what extent and in what forms are preverbal traumatic experiences retained in memory? and (3) Does trauma in the preverbal period have enduring effects? In these five preverbal infants, symptomatology consistent with typical posttraumatic diagnostic criteria was observed. The clinical material also suggested that the capacity to encode and retain meaningful internal representations of the salient elements of a traumatic experience may be present as early as the second half of the first year of life. The developmental implications of early trauma, particularly if it is severe, appear to be significant. PMID- 7480401 TI - The role of free movement in separation-individuation. A study of paralysis. AB - This paper reevaluates the role of free movement in the separation-individuation process through the special case of its absence. We put forward some ideas, formulated gradually during the psychoanalytically oriented psychotherapy of a paralyzed adult suffering from a narcissistic personality disorder. By analyzing the transference and countertransference in this case, we realized that the patient's inability to regulate his distance from his mother by free movement impelled him to use an alternative mode of regulating his object loss and re engulfment anxieties. We elaborate on two alternative mechanisms to cope with these anxieties: the motoric mode of approaching-distancing, and the mode of idealization-devaluation, inflating and deflating the inner significance of the object. We discuss the implications of these modes for the stability of self and object experiences, their different use of the Doing and Being experiential modalities, and their relation to pathological narcissism. PMID- 7480402 TI - Erotic transference in the male adolescent-female analyst dyad. AB - Erotic transference involving female analysts and adult male patients has received increased attention in the past ten years. Absent from the literature, however, are any clinical reports concerning erotic transferences of adolescent male patients with female therapists. This paper presents a fragment of a treatment involving an adolescent male patient who developed an erotic transference. The vicissitudes of the transference, as well as his relationship to his mother and father, are discussed from the perspective of developmental conflicts and tasks of adolescence. The countertransference dimensions of this particular gender constellation are also examined from the standpoint of intrapsychic and cultural factors. PMID- 7480403 TI - Daughters and mothers. Aspects of the representational world during adolescence. AB - This paper explores the transformations of the girl's intrapsychic relationship to the mental representations of her mother and their contribution to mastery of the specifically female developmental milestones during adolescence and early adulthood. The vicissitudes of the girl's intrapsychic organization of the mental representations to the maternal object both reflect and characterize the psychological tasks of female adolescence: puberty, menarche, sexuality, and psychological independence. It is argued that the characteristic fantasies of this period can be understood not only as representing an intrapsychic developmental progression along object lines but as complex compromise formations which serve multiple structural functions as well. Clinical material is presented to illustrate typical fantasies of female adolescence. PMID- 7480404 TI - Termination in child analysis. A child-led process? AB - The paper begins by drawing attention to the disjunction between the theoretical conceptualizations of termination in child analysis and the process as it actually occurs. The central focus is on the process itself. Material is presented from four child analyses to illustrate the idiosyncratic ways in which the termination phase unfolded. The cases selected make it possible to consider termination in relation to each developmental phase. Our central argument is that, although the analyst should have certain criteria for termination in mind, termination in child analysis is essentially a child-led process. PMID- 7480405 TI - The analysis of an overstimulated child. AB - This paper shows how environmental and medical overstimulation affected a latency girl in all her developmental phases, leading to severe impairment of object relationships. These factors are seen against the background of an acrimonious parental divorce. Suzie's negative body image was inextricably interwoven with anxieties about her masturbation activities and castration fantasies and this was fueled by her physical condition (lichen sclerosis et atrophicis). She was in a constant state either of withdrawness or excitement. The initial task of the treatment was to provide Suzie with a holding environment so that she could find alternative means of gratification and control in order to manage within a more organized secondary process mode. When her difficulties around excitation abated, she could begin to make analytic use of verbalization, clarification, and interpretations. PMID- 7480406 TI - Enactment and play following medical trauma. An analytic case study. AB - This paper on the four-year analysis of a five-year-old girl with a school phobia details the playroom enactments of medical trauma and its sequelae. Early in the analysis the patient expressed herself almost exclusively through play. Gradually she connected her conflicts, including those over her compulsive masturbation, to her hospital experiences. Only after the traumatic medical experiences were no longer the major issue did other important conflicts become available for analytic work. Physical activity remained prominent in the analysis and generated pressure for mutual enactment. This case again draws our attention to the complex overlap of play, enactments, and verbalization in child analysis. PMID- 7480407 TI - Post-traumatic stress and coping in an inner-city child. Traumatogenic witnessing of interparental violence and murder. AB - Violence today appears to be ubiquitous: it even enters the clinical session, deeply internalized within child victims who were exposed to often unspeakable horror. Violence and its pernicious, horrific effects are observed in the streets, schools, parks, playgrounds, and homes of some inner-city communities. This article introduces the use of Anna Freud's Diagnostic Profile system with an inner-city child who, at the age of four, witnessed his mother fatally stab his father with a kitchen knife and at age eleven was assessed and treated by the author. Clinicians may wonder whether any kind of therapy could ever undo the serious fixations, regressions, developmental arrests, and integrate trauma shattered ego functions observed in children exposed to visual horror and affective terror. Application of the Profile may offer some direction with these children: a panoramic view of their painful mood, their hypervigilance and distrust, fears, separation and annihilation anxieties, nightmares (with murder imagery), developmental anomalies and arrests is presented with clarity and force. The therapist uses countertransference responses to monitor the affect tolerance in the child and to determine the appropriate dosages of awareness the child can integrate from one moment to the next. The therapist also serves as the child's external stimulus barrier and explores feelings about media-driven portrayals of violence, stereotypes, and inner-city children and youths. The unsurpassed utility of the Profile as a diagnostic system that documents vital economic, dynamic, structural, genetic and adaptive-coping information about the child is discussed in detail as is the Profile's added benefit of possibly guarding against misdiagnosis and charting a course for psychotherapy in difficult city-violence trauma cases. PMID- 7480408 TI - Freud's psychology. Can it survive? AB - The increasing diversity of, and disparities between, psychoanalytic theories raises the question of whether, and to what extent, these various formulations remain firmly rooted in Freud's basic psychoanalytic concepts. Many analysts believe that these theories exemplify "unity in diversity." This view is contested in this paper. It is argued that Freud's basic concepts are his metapsychological concepts which, although capable of modification in the light of fresh clinical and theoretical findings are, in their fundamentals, indispensable. It is contended that it is precisely these fundamentals that are dislodged or put at risk by a great deal of current psychoanalytic thinking, and that it is in this (negative) respect that many diverse approaches are in concert. The argument is pursued with special reference to infant observational research, contemporary affect theory, and the neuroscience of Luria. PMID- 7480409 TI - Helping foster children to mourn past relationships. AB - In spite of the increasing number of foster children who have had pathological relationships with their biological parents, there are relatively few reports of intensive therapy with these children. This paper focuses on the psychoanalytic psychotherapy of two young foster children whose treatment began shortly after placement. The paper describes the children's developmental problems, their conflicts and defenses, and the therapeutic process utilized to enable them to resume progressive development. The children were helped to mourn their past objects and to form more appropriate attachments to their foster parents. PMID- 7480410 TI - Resistance analysis and psychic reality. AB - The purpose of this paper is to elucidate some aspects of our mode of working; how we as analysts listen, observe, and collect information to formulate appropriate resistance interventions. Analytic exploration and technique should to a greater extent than what is usual include a careful analysis of verbal and nonverbal resistance expressions, as these may contain important material related to the analysand's psychic and historical reality. Our theoretical and technical conceptualization of how a systematic and consistent resistance analysis is performed should be improved. Our listening and observing attitude involves the simultaneous use of various forms of auditory, visual, and enactive data. This is exemplified by an excerpt from an analytic hour in which a specific resistance problem--an analysand's suppressed response to crying--is examined and linked to specific preverbal experiences. PMID- 7480411 TI - Silent eulogy. Elective mutism in a six-year-old Hispanic girl. AB - A case of elective mutism in a six-year-old Hispanic girl treated in a public mental health setting is presented. The central feature of the case was the facilitation of a delayed grieving process in a little girl who had witnessed her father's death. The presentation stresses the critical need to integrate the unfolding clinical material with the realities of the stresses of parental distress and resistance. An attempt is made to understand this "resistance" in the context of cultural and social realities, while attending to the possible underlying psychodynamic meaning. PMID- 7480412 TI - Traumatic memory and the intergenerational transmission of Holocaust narratives. AB - This paper investigates the roles of affect regulation, narrative cohesion, and symbolic representation in the intergenerational transmission of the Holocaust experience. A study of the reminiscences of mothers who are Holocaust survivors and their daughters' reflections about the Holocaust illustrates the process of the transmission of trauma by tracing the transgenerational evolution of narrative forms, dynamic themes, and affective organization. The quality of the survivor parent's organization and integration of affect has significant bearing on how her child assimilates her knowledge of the Holocaust and develops the capacity to tolerate and express painful emotions. Through the preservation, transformation, and transmutation of traumatic memory, children of survivors strive to assimilate, redeem, and transform their tragic historical legacy. PMID- 7480413 TI - Models of reflexive recognition. Wallon's Origines du caractere and Lacan's "Mirror stage". AB - Jacques Lacan drew on the empirical research of the psychologist Henri Wallon both in his 1938 theory of the mirror stage ("the intrusion complex") and in subsequent formulations. Yet, like Lacan's famous "return" to Freud, his recourse to Wallon is revisionary and, at times, antithetical. This essay examines the disparities between Wallon's and Lacan's work on the mirror experience, focusing especially on the status of the mirror and the identity of the reflected image. An analysis of these differences helps to clarify the meaning of the child's specular activity--"an ontological structure of the human world"--in Lacan's later view and the implications of reflexive recognition for his conception of parental agency. PMID- 7480414 TI - The child is father of the man. Wordsworth's Ode: Intimations of immortality and his secret sharers. AB - William Wordsworth's "Ode: Intimations of Immortality" is manifestly about both the poet's loss of inspirational perceptive powers and emotional intensity with age, and the compensations of maturity. It also refers to the poet's fear that he might lose his "secret sharers," real or fantasied, consciously or unconsciously conceived parent substitutes for whom and with whom one creates. Wordsworth anticipated that with his upcoming marriage he would lose his sister Dorothy and his close friend and collaborator Samuel Taylor Coleridge, who played important roles in his creativity. Optimism and relief replaced sadness when he realized that he was not deprived of his sharers. The concepts of "secret sharers" and "collective alternates" for whom one creates are intimately related. PMID- 7480415 TI - "Lazarus stand forth" H. D. encounters Freud. AB - H.D., the noted writer Hilda Doolittle, wrote three differing accounts of her first analytic session with Freud, held on March 1, 1933. The first account was written on the same day as her session in a letter to her friend Bryher; it indicates that Freud did not meet H. D.'s preformed transference expectation. Although H. D. focused on her fear of Freud's rejection, her deeper concern was her disappointment in Freud and her fear of loving a person who might die soon. In 1944, five years after Freud's death, H. D. wrote a second account of the session in "Tribute to Freud" in which she attempted to recall an idealized memory of Freud. To do this her description split the session into a part in which she projected her anger onto Freud and a part in which she reconciled with Freud in a mystical, wordless communication by means of her relation to his dog, Yofi. Her third account, "Advent," was written in 1948, following a period of psychosis. At that time H. D. was able to acknowledge that she had been frightened of loving a man who was vulnerable to death and to restore Freud's presence by identifying with this analytic ability. PMID- 7480416 TI - The examination dream revisited. A clinical note. AB - In each of three dreams reported to me fortuitously within a few days of each other, the patient was anxious about a forthcoming medical examination. Two of the patients reported a sense of guilt for immoral behavior; the feared illness could be interpreted as punishment. PMID- 7480417 TI - Loewald's "Positive neutrality" and the affirmative potential of psychoanalytic interventions. AB - Patients who suffer from severe vulnerability in the narcissistic realm but who otherwise enjoy impressively intact ego functioning can sometimes be helped to endure the rigors of psychoanalysis in the early months of treatment by the analyst's emphasis on, or even exaggeration of, the affirmative elements in ordinary psychoanalytic interventions. Loewald's (1960) concept of "positive neutrality" is explored in an effort to determine an acceptable element of affirmation in the initial analytic stance taken toward these patients, and case material is offered to illustrate the application of these principles in the first fifteen months of psychoanalytic work with a high-functioning but narcissistically vulnerable man. It is thought that this stance, used judiciously, does not compromise the full emergence and analysis of the transference but rather enhances the process. To illustrate this progression, a session is presented from the thirtieth month in the treatment of the same analysand, when the affirmative emphasis had receded and more ordinary interpretive activity had begun to dominate the analytic interchange. PMID- 7480418 TI - A childhood gender identity disorder. Analysis, preoedipal determinants, and therapy in adolescence. AB - This paper is about a four-year-old boy's wish to be a girl. This wish was a compromise formation, the consequence of intrapsychic conflict. Using reconstructive inferences and research findings, I suggest a line of development in the boy's femininity, beginning with constitutional and maturational factors, with being shaped in a feminine direction by his environment, and with shaping himself internally in the form of a compromise formation in that direction by the time he was two to three years of age. The disorder began to acquire a persistence and stability during the preoedipal phase and, with minor changes in its components, remained operative during the oedipal phase, latency, and adolescence. PMID- 7480419 TI - Touching limits in the analytic dyad. AB - The matter of limits and boundary violations by both parties in the analytic dyad remains an unsettled technical and ethical concern, whether touching has to do with actual physical contact or is expanded in its meaning to include its psychic equivalents. Touching, probing, and breaching of the idiosyncratic perimeters of the private self of one by the other in the dyadic intimacy are necessary components of the healing contact but pose an inevitable liability for violation, disruption, and damage. Clinical data remind the analyst of the near-physical impact of words. And the data sometimes speak for the legitimate place of restrained forms of physical contact, as nonverbal necessities of analytic communication, in critical instances in which a viable analytic engagement could not otherwise be sustained. PMID- 7480420 TI - The ideal of the anonymous analyst and the problem of self-disclosure. AB - The principle of analytic anonymity is critically reviewed. A connection between the technical stance of nondisclosure and idealization of the analyst is proposed. Some preliminary suggestions are offered concerning what kinds of information about the analyst are useful to communicate to a patient. PMID- 7480421 TI - Aloneness in the countertransference. AB - This survey of a variety of ways in which analysands induce a feeling of aloneness in the analyst includes reference to the developmental origins, unconscious dynamics, and characterological settings of these ways. An account is presented of the role of narrative choice in defining the phenomena of an analysis not just in reporting pre-existing phenomena. Also included are some technical suggestions. PMID- 7480422 TI - The nature and function of a pathological oedipal constellation in a female patient. AB - The first analysis of a female patient dealt essentially with separation individuation and narcissistic issues. During the second analysis a dynamic constellation of narcissistic character pathology, some psychopathic tendencies, perverse fantasies, and enactments emerged. Prominent was an unconscious fantasy of stealing the sexual organs of one parent to sexually satisfy the other parent. The etiology of this constellation and its transmutations in analysis are described. PMID- 7480423 TI - The dream is the guardian of sleep. AB - Freud's concept that the dream protects sleep is first applied to a clinical vignette involving combat veterans and then re-examined. Although conceived in the context of the topographical model, it is more compatible with the structural model. The mind produces a believed hallucination mimicking gratification of impulses that would lead to awakening. It is hypothesized that the dream develops in the neonate during the arousal of the REM state to protect sleep from burgeoning object-directed impulses. The clinical vignette suggests that traumatic dreams can use past trauma to insure current vigilance. Traumatic memories may symbolize fantasy complexes in dreams. PMID- 7480424 TI - A classic revisited: K. R. Eissler's "The effect of the structure of the ego on psychoanalytic technique". AB - Re-examining a classic paper by K. R. Eissler forty years after its publication recalled for the author his own "origins" in psychoanalysis in the same period, allowing a vivid context in which to view Eissler's contribution. Eissler's attempt to maintain a disciplined clinical position based on the structural theory as the foundation for psychoanalysis as a science, in the face of many proposed changes in both technique and theory in the decade after Freud's death, made many analysts refer to this paper as a kind of bible for the orthodox. This revisit attempts to explore the questions of how and why such a characterization of this paper has taken place. PMID- 7480425 TI - Clinical practice, measurement and information technology. PMID- 7480426 TI - The Psychogeriatric Assessment Scales: a multi-dimensional alternative to categorical diagnoses of dementia and depression in the elderly. AB - The Psychogeriatric Assessment Scales (PAS) provide an assessment of the clinical changes seen in dementia and depression. Principal components analysis and latent trait analysis were used to develop a set of scales to summarize these clinical changes. There are three scales derived from an interview with the subject (Cognitive Impairment, Depression, Stroke) and three from an interview with an informant (Cognitive Decline, Behaviour Change, Stroke). Results are reported on the reliability and validity of these scales using data from clinical samples in Sydney and Geneva and a population sample from Canberra. The scales were found to have excellent validity when judged against clinical diagnoses of dementia and depression and could distinguish Alzheimer's from vascular dementia. Cut-off points were developed to indicate correspondence between scale scores and clinical diagnoses. Percentile rank norms were developed from the Canberra population sample. The PAS is easy to administer and score and can be used by lay interviewers after training. It is intended for application both in research and in services for the elderly. PMID- 7480427 TI - Saccadic abnormalities in psychotic patients. I. Neuroleptic-free psychotic patients. AB - Most of the previous research reporting abnormalities of rapid re-fixation eye movements (saccades) in patients with schizophrenia has used patients receiving neuroleptic medication. In this study non-neuroleptically medicated schizophrenics were compared with other psychiatric patients using a variety of saccadic paradigms to determine the specificity of saccadic dysfunction. The patient groups consisted of schizophrenics (N = 18), bipolar affectives (N = 18), anxiety neurotics (N = 10) and normal controls (N = 31), none of whom had received neuroleptic medication for the preceding 6 months. Four behavioural paradigms, reflexive, predictive, remembered and ANTI were used to elicit saccades. The primary abnormality in the schizophrenic group was a significantly increased rate of distractibility in the ANTI (saccades made towards the target rather than in an opposite direction) and REM (saccades made prior to the imperative cue) paradigms. The major neuropsychological variable predictive of these errors was Wisconsin card sort perseverative errors. These data, in conjunction with findings from previous neurological research, would seem to provide converging evidence towards dysfunction of prefrontal cortex in schizophrenia. PMID- 7480428 TI - Saccadic abnormalities in psychotic patients. II. The role of neuroleptic treatment. AB - The effects of dopamine-antagonistic neuroleptic (NL) medication on saccadic eye movements were compared in matched groups of 40 NL-treated and 18 NL-free schizophrenic patients and in 18 NL-treated and 14 NL-free bipolar affective patients. Manipulation of the saccadic paradigm yielded data on four types of saccade: those reflexively elicited by novel stimuli (REFLEX saccades), those directed towards the remembered location of a target now extinguished (REM) or towards the location where a predictably alternating target is expected to appear (PRED), or ANTI saccades, directed away from the stimulus to the mirror image location. Extensive psychiatric, neurological and neuropsychological assessments were also carried out on all subjects. The saccades of NL-treated patients, regardless of diagnosis, were less spatially accurate than those of NL-free patients, with a greater tendency to fall short of the target when generated towards the locus of a mentally represented target. This effect was greatest with a predictably alternating target, especially during periods when target visibility was withdrawn, only a temporal cue remaining. This pattern of impairment which is also found in early stages of Parkinson's disease is likely to be due to deficiency of striatal dopamine. Its best clinical predictors were disease duration, and Webster-Parkinsonism scores. Failure to suppress reflexive saccades to the stimulus in the REM and ANTI paradigms were more closely associated with schizophrenia than with NL treatment and were best predicted by negative symptoms and Wisconsin perseverative errors, both of which are widely regarded as indicators of frontal lobe dysfunction. PMID- 7480429 TI - Concepts of mental illness and medical pluralism in Harare. AB - The Focus Group Discussions (FGD) described in this paper are the first step of a study aiming to develop an 'emic' case-finding instrument. In keeping with the realities of primary care in Zimbabwe, nine FGD were held with 76 care providers including 30 village community workers, 22 traditional and faith healers (collectively referred to as traditional healers in this paper), 15 relatives of patients and 9 community psychiatric nurses. In addition to the general facets of concepts of mental illness, three 'etic' case vignettes were also presented. A change in behaviour or ability to care for oneself emerged as the central definition of mental illness. Both the head and the heart were regarded as playing an important role in the mediation of emotions. The types of mental illness described were intimately related to beliefs about spiritual causation. Angered ancestral spirits, evil spirits and witchcraft were seen as potent causes of mental illness. Families not only bore the burden of caring for the patient and all financial expenses involved, but were also ostracized and isolated. Both biomedical and traditional healers could help mentally ill persons by resolving different issues relating to the same illness episode. All case vignettes were recognized by the care providers in their communities though many felt that the descriptions did not reflect 'illnesses' but social problems and that accordingly, the treatment for these was social, rather than medical. The data enabled us to develop screening criteria for mental illness to be used by traditional healers and primary care nurses in the next stage of the study in which patients selected by these care providers on the grounds of suspicion of suffering from mental illness will be interviewed to elicit their explanatory models of illness and phenomenology. PMID- 7480430 TI - Magnetic resonance imaging volumetric measurements of the superior temporal gyrus, hippocampus, parahippocampal gyrus, frontal and temporal lobes in late paraphrenia. AB - Quantified magnetic resonance measurements were made of superior temporal gyrus, parahippocampal gyrus, hippocampal, frontal and temporal lobe volumes and of the planar area of the thalamus and basal ganglia structures in 47 late paraphrenic patients and 33 healthy elderly controls. The late paraphrenics were divided into 31 schizophrenics and 16 patients with delusional disorder according to ICD-10 guidelines. Patients with delusional disorder tended to have smaller left temporal volumes compared with control subjects and patients with schizophrenia, but this difference failed to reach accepted levels of statistical significance after correction for the effects of multiple statistical comparisons, age and total brain size. Physiological right-left asymmetry, reported for temporal and frontal lobe volumes, was present in control, schizophrenic and delusional disorder subjects but delusional disorder patients had a significantly greater degree of temporal lobe asymmetry. The results add to the evidence for heterogeneity among late-onset psychoses and emphasize the subtle nature of any structural brain abnormalities in these patients. PMID- 7480431 TI - Neuropsychiatric and neuropsychological correlates of delusions in Alzheimer's disease. AB - We examined the prevalence, phenomenology, and clinical correlates of delusions in a consecutive series of 103 patients with probable Alzheimer's disease (AD). Patients were examined with the Present State Exam and the Dementia-Psychosis Scale. Twenty-one patients (20%) met DSM-III-R criteria for a delusional disorder. The most frequent delusion type was paranoid (71%), followed by hypochondriacal (67%), the Capgras syndrome (29%), house misidentification (29%), and grandiose delusions (29%). Out of the 21 AD patients with delusions, 76% had three or more different types of delusions simultaneously. The frequency of delusions was not significantly associated with age, education, or age at dementia onset, and the type and severity of cognitive impairments was similar for AD patients with and without delusions. However, AD patients with delusions had significantly higher mania and anosognosia scores. PMID- 7480432 TI - Insight and psychosis: a social perspective. AB - The last 5 years have seen a resurgence of interest in insight, with better definitions and operationalizations of the concept. However, recent work has paid little attention to the ways in which patients' understanding of their illnesses may be shaped by perceptions of mental illness and treatment prevailing in their cultures and social groups. We review work by social scientists on cultural and social variations in lay perceptions of mental illness and on the stigma attached to mental illness, and argue that these varying views are likely to be one of the influences on insight, requiring further exploration in research on this subject. PMID- 7480433 TI - The structure of common psychiatric symptoms: how many dimensions of neurosis? AB - In order to replicate and elaborate the two-dimensional model of depression and anxiety underlying the structure of common psychiatric symptoms proposed by Goldberg et al. (1987), we carried out latent trait analyses on PSE symptom data of the original Manchester study and two recent Dutch studies. We used the same analytical strategy as Goldberg et al. to facilitate comparison with the earlier work. It was found that a more comprehensive set of common psychiatric symptoms caused an extra, third dimension to emerge, so that the earlier anxiety dimension became split between a specific anxiety axis characterized by situational and phobic anxiety and avoidance, and a non-specific anxiety axis characterized by free-floating anxiety, various symptoms relating to tension, irritability and restlessness. It is argued that three dimensions are sufficient to account for the covariance between common psychiatric symptoms. A fairly consistent correlation between the non-specific anxiety and the depression dimension was found across sites, as well as independence of the specific anxiety dimension from the other two dimensions. Furthermore, the depression dimension was robust with similar symptom profiles across samples, but there appeared to be local differences in the structure of anxiety symptoms. PMID- 7480434 TI - Exclusion of linkage of schizophrenia to the gene for the dopamine D2 receptor (DRD2) and chromosome 11q translocation sites. AB - There have been previous reports of a 1q43;11q21 translocation cosegregating with schizophrenia and a 9p22;11q22.3 translocation cosegregating with manic depression. In addition, the genes for the dopamine D2 receptor and for tyrosinase both map to chromosome 11q. Three 11q DNA markers were used to investigate 23 pedigrees containing multiple cases of schizophrenia. Strongly negative lod scores were obtained, providing evidence against linkage over a 70 cM region which included both translocation sites and both candidate genes. PMID- 7480435 TI - Psychosocial sequelae of the 1989 Newcastle earthquake: I. Community disaster experiences and psychological morbidity 6 months post-disaster. AB - A stratified random sample of 3007 Australian adults completed a screening questionnaire 6 months after the 1989 Newcastle earthquake. Information was obtained on initial earthquake experiences and reactions, use of specific services, social support, coping strategies and psychological morbidity. This questionnaire was the first phase of the Quake Impact Study, a longitudinal project investigating the psychosocial impact of the earthquake. Two weighted indices of exposure were developed: a threat index, which measured exposure to injury or the possibility of injury; and a disruption index, which measured experiences of property damage, displacement and other losses. Levels of exposure to threat and disruption events were significant predictors of morbidity on both the General Health Questionnaire and Impact of Event Scale, as were coping style and gender. Effects of exposure to threat and disruption were largely additive, with higher exposure being associated with greater use of support services, higher perceived stressfulness and more severe psychological morbidity. Use of avoidance as a coping strategy, female gender, lower social support and being older were also associated with higher post-disaster psychological distress. It was estimated that 14.8% of the population was exposed to high levels of threat or disruption, of whom approximately 25% experienced moderate to severe psychological distress as a direct result of the disaster. It was further estimated that 18.3% of those exposed to high levels of threat were at risk of developing post-traumatic stress disorder, representing approximately 2% of the city's adult population. PMID- 7480436 TI - Lateralized abnormality of high energy phosphate metabolism in the frontal lobes of patients with bipolar disorder detected by phase-encoded 31P-MRS. AB - High energy phosphate metabolites were measured using phase-encoded in vivo phosphorus-31 magnetic resonance spectroscopy (31P-MRS) in both the left and right frontal lobes of 25 patients with bipolar disorder. Eleven patients were examined in the depressive state, 12 in the manic state, and 21 in the euthymic state. Twenty-one age-matched normal volunteers were also examined. The phosphocreatine (PCr) peak area percentage in the left frontal lobe in the patients in the depressive state was decreased compared with that in the normal controls. It was significantly negatively correlated with the Hamilton Rating Scale for Depression score evaluated at the time of 31P-MRS examination. The PCr peak area percentage in the right frontal lobe in the patients in the manic and the euthymic states was decreased compared with that in the controls. These results are compatible with previous reports describing reduction of glucose metabolism in the left frontal lobe in depressive patients with bipolar disorder and trait-dependent right hemisphere dysfunction in bipolar disorder. PMID- 7480437 TI - Social, psychological and physical correlates of eating problems. A study of the general adolescent population in Norway. AB - The characteristics and concurrent predictors of eating problems were identified. Ninety-seven per cent of a representative sample of Norwegian adolescents (N = 11315) completed a questionnaire containing a 12-item version of the Eating Attitudes Test (EAT) and measures of a large number of physiological, psychological and social factors previously suggested to be of aetiological importance. Social class, drug use and indicators of 'the model child' (high grades, high occupational aspirations and much homework) were not related to eating problems. Logistic regression analyses identified perceived obesity as the strongest associated factor, followed by gender, depression, excessive exercise and unstable self-perceptions. In addition, adopting idols with perfect bodies, body dissatisfaction, low self-worth, feminine sex-role orientation, lack of parental care, early pubertal timing, age and number of alcohol intoxications all added to the probability of eating problems. About 6% perceived themselves as obese in spite of subnormal BMI. Such misperceptions increased the risk of high scores on EAT. PMID- 7480438 TI - The neuropsychology of schizophrenia: relations with clinical and neurobiological dimensions. PMID- 7480439 TI - People with learning disabilities and challenging behaviour: the characteristics of those referred and not referred to specialist teams. AB - The characteristics of two samples of people with learning disabilities and severely challenging behaviour were examined. The experimental sample comprised subjects who had been referred to two specialist challenging behaviour services in separate counties. The comparison sample comprised subjects from the same counties who were not referred or accepted for specialist intervention. No significant differences were found between the samples in terms of age, gender, social impairment, dependency characteristics, ability levels and mental health. Significant differences between them were found in the nature and severity of aberrant behaviour presented, as assessed by their carers. The implications of these findings are discussed with regard to how the term 'severely challenging behaviour' is defined in practice. PMID- 7480440 TI - The Cardinal Needs Schedule--a modified version of the MRC Needs for Care Assessment Schedule. AB - This paper describes a modified version of the MRC Needs for Care Schedule (the Cardinal Needs Schedule), for measuring needs for psychiatric and social care amongst patients with severe psychiatric disorders. The modified schedule has three new features: (i) it is quick and easy to use; (ii) it takes systematic account of the views of patients and their carers; (iii) it defines and identifies need in a way that is concise and easy to interpret. The paper describes why the three new features were considered necessary, and then gives an overview of the structure of the Cardinal Needs Schedule, together with a description of how the three new features were developed. During a study of social services care management the practicality of the modified schedule was investigated and further data were obtained on the reliability and validity of the standardized approach to measuring need, in domains not previously investigated. Because of its speed and simplicity, the Cardinal Needs Schedule offers a new choice to researchers who wish to use a standardized and practical assessment of need in evaluative studies of community care. Examples of the usage of the modified schedule are given in an Appendix. PMID- 7480441 TI - Neuropsychological evidence for frontostriatal dysfunction in schizophrenia. AB - Schizophrenics and controls were compared on a computerized test of attentional set-shifting which provides a componential analysis of the Wisconsin Card Sort Test and has previously been shown to be sensitive to frontal lobe dysfunction and Parkinson's disease. The main test was of extra-dimensional shifting where subjects are required to shift response to an alternative perceptual dimension. In one condition, termed 'perseveration', subjects are required to shift to a novel dimension and ignore the previously relevant one. In the other condition, termed 'learned irrelevance', subjects are required to shift to the previously irrelevant dimension and ignore a novel one. Chronic medicated schizophrenics (N = 32) show a highly significant impairment on the perseveration but not the learned irrelevance condition, as compared to normal age and IQ matched controls (N = 24). This was true even of a subgroup of patients with preserved IQ. The impairments in attentional set-shifting failed to correlate with patients' scores on the Mini-Mental State Examination (mean; S.D. 26.8; 1.8) or with scores on a test of recognition memory. These results provide evidence for a specific deficit in a set-shifting test of executive function and support a hypothesis of frontostriatal dysfunction in schizophrenia. PMID- 7480442 TI - Electroencephalography and single photon emission computed tomography in dementia: a comparative study. AB - A series of elderly patients with dementia of Alzheimer type (AD), multi-infarct dementia (MID) and functional (non-organic) psychiatric illness (major depressive disorder) were selected according to DSM-III-R criteria and received: a battery of cognitive tests, EEG and Single Photon Emission Computed Tomography (SPECT) using 99mTc HMPAO. The EEG and SPECT scans were examined independently of the clinical data. The former were divided into two abnormal categories, those showing AD type change and vascular change respectively, and a normal group. The SPECT scans were classified as follows: a SPECT rCBF pattern showing bilateral temporoparietal perfusion deficits (AD type); those showing single focal perfusion deficits or multiple areas of low perfusion in the cerebral cortex suggestive of ischaemic change (vascular type SPECT picture); a mixed AD/MID pattern; and those with normal scan findings. There were significant associations between clinical diagnosis, EEG rating and SPECT rCBF pattern, approximately three-quarters of AD and MID patients having the predicted EEG and scan changes. Normal EEG recordings were more common in the MID patients. The two tests agreed in about two-thirds of cases, with no consistent pattern apparent in the cases with divergent findings. Each test misclassified a minority of dementia patients, but in only one patient were both investigations normal. Almost half of the so called 'functionally ill' patients had abnormal rCBF changes, showing mainly vascular changes while one-fifth had abnormal EEGs. PMID- 7480443 TI - Perspective production in a savant autistic draughtsman. AB - This study examines perspective construction in an autistic patient (E.C.) with quasinormal intelligence who exhibits exceptional ability when performing three dimensional drawings of inanimate objects. Examination of E.C.'s spontaneous graphic productions showed that although his drawings approximate the 'linear perspective' system, the subject does not use vanishing points in his productions. Nevertheless, a formal computational analysis of E.C.'s accuracy in an experimental task showed that he was able to draw objects rotated in three dimensional space more accurately than over-trained controls. This accuracy was not modified by suppressing graphic cues that permitted the construction of a vanishing point. E.C. was also able to detect a perspective incongruency between an object and a landscape at a level superior to that of control subjects. Since E.C. does not construct vanishing points in his drawings, it is proposed that his production of a precise realistic perspective is reached without the use of explicit or implicit perspective rules. 'Special abilities' in perspective are examined in relation to existing theoretical models of the cognitive deficit in autism and are compared to other special abilities in autism. PMID- 7480444 TI - Premenstrual syndrome. PMID- 7480445 TI - The anatomy of melancholia: does frontal-subcortical pathophysiology underpin its psychomotor and cognitive manifestations? PMID- 7480446 TI - Cognitive decline in an elderly population--a two wave study of change. AB - A sample of 1111 survivors from a population aged 75 years and over completed the Mini-Mental State Examination (MMSE) twice, separated by 28 months on average. There was a mean decline of 1.3 points in MMSE score. With increasing age, mean drop in score also increased. The proportion at each age identified as newly cognitively impaired according to any standard cut-point on MMSE rose markedly. Mean decline was greater in women than men even after adjustment for age. Cognitive change on the MMSE was approximately unimodally and normally distributed. This distribution was a marked contrast to the distribution of MMSE scores themselves, which was skewed due to truncation of scores at the maximum. The decline was not due to the inclusion of individuals with physical impairment. These findings indicate that cognitive decline, like dementia, becomes increasingly common with advancing age, and suggest that dementia may be regarded as one extreme of the continuum of cognitive decline. PMID- 7480448 TI - Clinical and cognitive diversity of psychotic states arising in late life (late paraphrenia). AB - This study aimed to explore the heterogeneity of paranoid psychosis with onset in late life by using cognitive factors in a centroid method of cluster analysis. Forty-seven subjects were allocated to two different clusters, the first with 24 (51.1%) and the second with 23 (48.9%) patients. Their cognitive attainment was evaluated against the performance of 33 elderly controls, all groups being matched for age, sex, and the numbers of years of education. Patients in cluster 2 showed a pattern of widespread cognitive impairment, which involved general measures of cognitive performance (MMSE, CAMCOG, WAIS-R verbal and performance scores), memory (digit and spatial span, delayed matching-to-sample, recognition memory for words and faces), and executive functions (verbal fluency, extra and intra-dimensional shift ability, spatial working memory, and planning). In contrast, patients in cluster 1 were only impaired on their extra-dimensional set shift and planning abilities, suggesting a more specific and restricted executive functioning deficit. We also analysed the impact that the use of antipsychotic medication could have had on patients' cognitive performance, which was shown to be negligible. In addition, there was no difference between the clusters with regard to the number of patients using neuroleptics, suggesting that the medication was unlikely to have introduced a performance bias in the two patient clusters. The validity of the subdivision of these patients into two separate groups was further supported by other clinical findings. Patients in cluster 1 exhibited more severe psychotic symptoms, as measured by the SAPS, than their counterparts in cluster 2, and were also more likely to display first-rank symptoms of Schneider. Conversely, cluster 2 membership was strongly associated with the presence of neurological signs and negative symptoms. We suggest that psychotic states arising in late life are a heterogeneous condition that may be best divided in two: 'type A', including patients with a wide range of psychotic symptoms, mild increase in the frequency of neurological signs, and cognitive deficits restricted to executive functions, and 'type B', which includes patients with less complex psychotic symptoms associated with a marked increase in the frequency of neurological signs and generalized cognitive impairment. The basis for this subdivision and the prospect for future studies are discussed. PMID- 7480447 TI - Cognitive features of psychotic states arising in late life (late paraphrenia). AB - The cognitive performance of 47 elderly psychotic patients with onset of symptoms in late life (late paraphrenia) was compared to that of 33 controls matched for age, sex, ethnic origin, number of years of education, and pre-morbid IQ as measured by the NART. Neuropsychological indices of general cognitive functioning (MMSE, CAMCOG, WAIS-R verbal and performance scores) showed that patients were performing the tasks at a significantly lower level than controls. Patients also showed a trend to have a lower span capacity than controls, particularly at the spatial span subtest. There was no obvious impairment of learning as measured by the digit and spatial recurring span tasks nor of simultaneous matching-to-sample ability. However, patients' performance on a delayed-matching-to-sample procedure was significantly worse than that of controls. In addition, patients performed worse than controls on the Recognition Memory Test for Faces, but not for Words. Finally, the performance of patients on tests assessing executive functioning (Verbal Fluency Test, Computerized Extra and Intra-Dimensional Shift Task, Computerized Spatial Working Memory Task, and Computerized Tower of London Task) was consistently worse than that of controls. These results suggest that psychotic states arising in late life are predominantly associated with a decline on measures of general cognitive ability and executive functioning. The neuropsychological meaning of these findings is discussed in the light of cognitive models of psychotic symptoms, as well as of schizophrenia and dementia research. We concluded that the lack of a clear pattern of impairment among these patients may be the result of their clinical and cognitive diversity. PMID- 7480449 TI - Three syndromes in schizophrenia: validity in young patients with recent onset of illness. AB - The heterogeneity of schizophrenia constitutes an impediment in the identification of reliable biological markers of the illness and aetiological factors. The two syndrome concept of the illness has proved a powerful stimulus in this endeavour. There is however evidence now to regard this construct as inadequate and in need of refinement. Using a sample of young patients with early onset of illness who fulfilled the Research Diagnostic Criteria for schizophrenia, we have shown that three rather than two dimensions better describe the pathology of the illness. The results of factor analysis showed these syndromes, termed disorganization, positive, and negative syndromes, to account for 65% of the total variance of the psychopathology of the disorder. The negative syndrome was associated with disturbed pre-morbid functioning while the disorganization syndrome bore significant association with early onset of illness, poorer socio-educational attainment, and impairment on neuropsychological tests, especially those designed to evaluate frontal lobe functioning. The positive syndrome was not associated with any of the assessed variables. These results provide strong support for the validity of the three syndrome construct even among a sample of non-institutionalized patients. PMID- 7480450 TI - The family life cycle: a forgotten variable in the study of women's employment and well-being. AB - One hundred and sixteen working-class housewives, part-time and full-time employees were compared on a range of well-being measures in three different lifestages. A group of factory employees was contrasted with a 'mixed occupation' group for a comparison of type of employment. The general contention that the association between employment and well-being would vary with position in the life cycle was confirmed for distress, physical illness and some of the satisfaction measures. Only one measure (personal competence) suggested that employment had a positive influence on well-being irrespective of lifestage. Young employees working for longer hours and older housewives were both defined as 'cases' by the General Health Questionnaire (GHQ-12). However, their profiles on the other well-being variables were very different. PMID- 7480452 TI - A test-retest reliability study of child-reported psychiatric symptoms and diagnoses using the Child and Adolescent Psychiatric Assessment (CAPA-C). AB - Seventy-seven 10-18-year-old psychiatric in-patients and out-patients took part in a test-retest study of the Child and Adolescent Psychiatric Assessment (CAPA). They were interviewed on two occasions several days apart. Overall reliability of diagnosis ranged from kappa = 0.55 (conduct disorder) to 1.0 (substance abuse or dependence). In general, reliability for scale scores of psychopathology was somewhat lower in out-patients than in-patients, though the opposite was the case for anxiety disorders and psychosocial incapacity and the reliability of the diagnosis of conduct disorder--the only individual diagnosis sufficiently common to permit this comparison. Unreliability of reports of behavioural problems was found to be related to admitting to being a liar in the first interview. The implications of these results for the use of the CAPA are discussed. PMID- 7480451 TI - The Child and Adolescent Psychiatric Assessment (CAPA). AB - Great advances have been made during the last 20 years in the development of structured and semi-structured interviews for use with psychiatric patients. However, in the field of child and adolescent psychiatry there have been weaknesses in the specification and definition of both symptoms and the psychosocial impairments resulting from psychiatric disorder. Furthermore, most of the available interviews for use with children have been tied to a single diagnostic system (DSM-III, DSM-III-R, or ICD-9). This has meant that symptom coverage has been limited and nosological comparisons have been inhibited. The Child and Adolescent Psychiatric Assessment (CAPA) represents an attempt to remedy some of these shortcomings. This paper outlines the principles adopted in the CAPA to improve the standardization, reliability and meaningfulness of symptom and diagnostic ratings. The CAPA is an interviewer-based diagnostic interview with versions for use with children and their parents, focused on symptoms occurring during the preceding 3 month period, adapted for assessments in both clinical and epidemiological research. PMID- 7480453 TI - A prospective clinical study of stereotactic subcaudate tractotomy. AB - This study describes a cohort of 23 patients undergoing stereotactic subcaudate tractotomy. Research Diagnostic Criteria indicated that 70% suffered major depressive disorder; the remainder mostly had a bipolar affective disorder. There were serial assessments pre-operatively and at 2 weeks and 6 months post operatively using the Hamilton Rating Scale for depression, the Present State Examination (PSE), Newcastle Scale, the Beck Depression Inventory, and the Taylor Manifest Anxiety Scale. Neuropsychological assessment included tests thought to be sensitive to frontal lobe dysfunction, as well as tests of general intelligence, attention, memory, language and visuo-spatial function. Post operatively, depression rating scale scores decreased significantly but most patients continued to exhibit a number of PSE syndromes. Depression rating scale scores were correlated with 1 year global outcome: there was no significant correlation except for the 6 month assessment when lower Hamilton scores were found to be associated with better global outcome. Correlations between the neuropsychological tests and the Hamilton and Beck depression scales at 2 weeks post-operatively suggested that an improvement in psychiatric condition was associated with greater efficiency on some tests of attention and verbal recall, as well as faster performance on a sorting task. By contrast, the changes at 6 months suggested an association between improvement in psychiatric condition and less efficient performance on certain neuropsychological tests including verbal recognition memory, attention and two tests of frontal lobe dysfunction. PMID- 7480454 TI - Prescription pattern of antidepressants in out-patient psychiatric practice. AB - The prescription pattern of antidepressants was reviewed in a large sample of psychiatric out-patients in the Region of Piedmont. Antidepressants were administered to 780 patients; most were diagnosed with mood disorders (62% of prescriptions) and anxiety disorders (34%), the rest with other diagnoses. The mean prescribed dose of antidepressants was at the lower end of, or below, the recommended range for out-patients, with the exception of amineptine and mianserin. The administered dose of tricyclic antidepressants was significantly associated with sex and diagnosis: female patients received a daily dose that was significantly lower than that for males, and patients with a diagnosis of mood disorders received significantly more medication than those with other diagnoses. PMID- 7480455 TI - The Gospel Oak Study stage IV: the clinical relevance of subjective memory impairment in older people. AB - The prevalence rate of subjective memory impairment (SMI) and its value as a predictor of future depression or dementia was studied in a community sample of elderly residents in one electoral ward using the short-CARE. SMI was found to be common, occurring in 25% of subjects. Subjects with SMI were more likely to be suffering from either dementia or depression than those without the complaint, although 60% of subjects with SMI did not have evidence of either disorder. When followed up over a 2-year period, subjects with SMI were found to be at four-fold greater risk of developing future dementia and two-fold greater risk of developing a depression compared with those without SMI. The SMI scale was not found to be useful as a population screen for dementia or depression, although two of the nine items might have value as screening questions in clinical circumstances to determine those with memory complaints at risk for dementia. PMID- 7480456 TI - The relationship between bipolar disorder and alcoholism: a controlled family study. AB - Bipolar disorder and alcoholism are familial disorders. The familial-genetic relationship between both is controversial and has received insufficient study. This study explores whether bipolar disorder and alcoholism share familial risk factors, and whether the co-occurrence of lifetime diagnosis of bipolar disorder and alcoholism is familial. We report on first-degree relatives of 146 consecutively admitted patients with either bipolar disorder or/and alcoholism; relatives of the patients (in total 728 relatives directly interviewed) were compared with first-degree relatives of 109 general population probands (320 relatives directly interviewed). Overlap between the familial components underlying bipolar disorder and alcoholism was not observed if the analysis was restricted to 'pure' diagnostic groups. Excess comorbidity between bipolar disorder and alcoholism was observed in relatives. Multiple sources for this excess of comorbidity between major affective disorders and alcoholism in families of probands with bipolar disorder are likely; in particular, we found evidence for a distinct subgroup of comorbid cases with familial comorbidity; however, excess comorbidity was also found in absence of familial loading with alcoholism. PMID- 7480457 TI - Familial depression versus depression identified in a control group: are they the same? AB - Subjects who meet the criteria for an affective syndrome possibly are aetiologically heterogeneous. An approach to this possibility involves examining affectively ill subjects obtained by different methods of ascertainment. This study compares depressed and manic subjects who are related to affectively ill probands with affectively ill subjects who were obtained from a study of a control population, and, therefore, were less likely to be familial. The subjects were identified in a large collaborative study of depression where both family members as well as controls were personally interviewed and followed up for 6 years after admission to the study. Data were obtained on subtypes of affective disorder using the Research Diagnostic Criteria and information was gathered about psychiatric hospitalizations, suicide attempts, alcoholism and psychological functioning prior to admission. Similar assessments were made for the comparison groups for the 6 year period between intake and follow-up. Relatives of bipolar I/schizoaffective manic probands were more likely to show mania than affectively ill controls or relatives of unipolar/schizoaffective depressed probands. Affectively ill controls were less likely to be hospitalized and less likely to suffer from an incapacitating depression. They were also likely to have functioned in a more healthy fashion than the affectively ill relatives of the bipolars and unipolars, in the 5 years before admission to the study. In the 6 year follow-up, both the subjects themselves and raters assessed the depressed controls as functioning better than the affectively ill relatives of the probands. Further, assessment of global adjustment during the 6 year period was worse for the relatives of affectively ill probands than for the depressed controls. Length of major depression was longer in relatives of bipolar and unipolar probands than in controls. Though all of the subjects in this study met research criteria for an affective illness, there were marked differences in the qualitative aspects of these illnesses with the relatives of affectively ill probands, who functioned less well and had longer and more severe episodes and more hospitalizations. PMID- 7480458 TI - Is seeking treatment for depression predicted by a history of depression in relatives? Implications for family studies of affective disorder. AB - Most family studies of psychiatric illness ascertain probands through treatment facilities. If psychiatric illness in relatives influences the probability of treatment seeking, then the risk of illness in the relatives of treated probands will not be representative of the risk of illness in all probands. We have investigated this question in a population-based sample of female twins with a lifetime history of DSM-III-R defined major depression (MD) (N = 753), 36% of whom reported seeking professional help for their depression. Logistic and Cox regression methods were used. Treatment seeking for MD was significantly associated with age, education, comorbidity with anxiety disorders, degree of impairment and number of symptoms. Controlling for these predictors, treatment seeking was significantly predicted by the presence of one or more relatives with a lifetime diagnosis of MD (OR = 1.62). The probability of treatment seeking for MD in the proband was significantly increased if the affected relative: (i) had an onset of their MD prior that of the proband and; (ii) themselves sought treatment for their depression. Seeking treatment in this epidemiological sample of depressed women was associated with a 32% increase in risk of MD in relatives. Selecting probands for family studies from treated populations may select for families with an increased rate of illness. For MD, this artefact can explain only a modest proportion of the familial aggregation observed in prior family studies that utilized treated probands. PMID- 7480459 TI - Sub-typing depression, I. Is psychomotor disturbance necessary and sufficient to the definition of melancholia? AB - Melancholia is most commonly distinguished from non-melancholic depression by the presence of psychomotor disturbance (PMD) and a set of 'endogeneity' symptoms. We examine the capacity of an operationalized clinician-rated measure of PMD (the CORE system) to predict diagnostic assignment to 'melancholic/endogenous' classes by the DSM-III-R and Newcastle systems. Examining a pre-established CORE cut-off score (> or = 8) against independent diagnostic assignment, PMD was present in 51% of those assigned as melancholic by DSM-III-R, and 85% of those assigned as endogenous by the Newcastle system, quantifying the extent to which it is 'necessary' to the two definitions of 'melancholia'. Additionally, multivariate analyses established that the addition of a refined set of historically suggested endogeneity symptoms added only slightly to overall discrimination of melancholic and non-melancholic depressives. While only few endogeneity symptoms independent of psychomotor disturbance were suggested, their specific relevance varied against system definition of melancholia (appetite/weight loss and terminal insomnia being identified for DSM-III-R; anhedonia for Newcastle; and diurnal variation in mood and energy for both systems). Results allow consideration of the relative importance of two domains (psychomotor disturbance and 'endogeneity' symptoms) to clinical definition of melancholia, and have the potential to assist both classification and pursuit of neurobiological determinants. We interpret findings as suggesting a 'core and mantle' model for conceptualizing the clinical features of melancholia, with psychomotor disturbance as the core and with independent endogeneity symptoms as only a thin mantle. PMID- 7480460 TI - Sub-typing depression, II. Clinical distinction of psychotic depression and non psychotic melancholia. AB - We have attempted to clarify clinical differentiating features of psychotic depression. Forty-six depressed subjects meeting DSM-III-R criteria for major depression with psychotic features were compared with (i) DSM-defined melancholic, (ii) Newcastle-defined endogenous, and (iii) a residual DSM-defined major depressive episode group. Additionally, a 'bottom up' latent class analysis (LCA) suggested a larger sample of 82 'psychotic depressive' subjects, and multivariate analyses contrasted these subjects with both LCA-identified melancholic and all residual depressed subjects. Analyses suggested that, in addition to two features with absolute specificity (delusions and hallucinations), both the DSM-defined and LCA-defined 'psychotic depressive' subjects were significantly more likely to demonstrate marked psychomotor disturbance, to report two morbid cognitions (feeling sinful and guilty; feeling deserving of punishment), as well as be more likely to report constipation, terminal insomnia, appetite/weight loss and (variable across the defined 'psychotic depressive' groups) loss of interest and pleasure. The study identifies a wider set of potentially discriminating clinical variables than previous studies, as well as both indicating the existence and assisting identification of 'true' psychotic depression in the absence of formal psychotic features being acknowledged or elicited. PMID- 7480461 TI - Sub-typing depression, III. Development of a clinical algorithm for melancholia and comparison with other diagnostic measures. AB - We describe the development of a clinical algorithm to differentiate melancholic from non-melancholic depression, using refined sets of 'endogeneity' symptoms together with clinician-rated CORE scores assessing psychomotor disturbance. Assignment by the empirically developed algorithm is contrasted with assignment by DSM-III-R and with several other melancholia sub-typing indices. Both the numbers of 'melancholics' assigned by the several systems and their capacity to distinguish 'melancholics' on clinical, demographic and a biological index test (the DST) varied across the systems with the algorithm being as 'successful' as several systems that include inter-episode and treatment response variables. Analyses provide information on the criteria set developed for DSM-IV definition of 'melancholia'. PMID- 7480462 TI - Three-year follow-up of Mini-Mental State Examination score in community residents aged 85 and over. AB - The objective of this study was to describe over time the course of cognitive function of elderly without cognitive impairment and of elderly with different stages of impairment, and to assess if the change in cognitive function was dependent on the initial level of function. The Mini-Mental State Examination (MMSE) was used at two time points. The first assessment (MMSE-1) was part of a community-based study and was obtained from 871 subjects. For the second assessment (MMSE-2) a sample of 166 subjects was drawn from the subjects alive at follow-up who had an MMSE-1 score. This sample was stratified by MMSE-1 score to avoid oversampling of subjects with high MMSE-1 scores. A second MMSE score was obtained from 134 elderly, whereas 18 subjects refused participation and 14 subjects were not traceable. The median age at first assessment was 89 years (25th percentile 87, 75th percentile 92), the mean follow-up period (S.D.) was 3.3 (0.5) years. The median change in MMSE score was minus 4 points (95% confidence interval (CI) -7 to -2) and the slope of the regression line of MMSE-2 on MMSE-1 was 1.1 (95% CI 0.9-1.3). It is likely that the slope was underestimated due to a floor effect, regression to the mean and missing observations. However, the probability of decline decreased if MMSE-1 was higher. Nevertheless, the probability ranged from 27 to 59% for subjects with the highest MMSE-1 scores aged 85 and 95 years respectively.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7480463 TI - Sequential cross-sectional and 10-year prospective study of severe negative symptoms in relation to duration of initially untreated psychosis in chronic schizophrenia. AB - Current clinical correlates of duration of initially untreated psychotic symptoms were investigated in a cross-sectional analysis followed by a 10-year prospective study among 88 in-patients with a long-standing schizophrenic illness, many of whom had experienced prolonged periods of untreated psychosis due to illness onset and hospital admission in the pre-neuroleptic era. After controlling for the effects of age, and duration and continuity of subsequent neuroleptic treatment, the primary clinical correlate of duration of initially untreated psychosis was muteness. Over the subsequent 10-year-period, no new cases of muteness emerged and some existing cases of muteness partially resolved, though the speech that emerged remained very sparse and revealed generally gross cognitive debility. The pathophysiology underlying active, unchecked psychosis may also constitute an active morbid process that is associated with the further progression of severe negative symptoms and cognitive dysfunction in the long term. PMID- 7480464 TI - Schizophrenic subjects with no history of admission to hospital. AB - It has often been assumed that all subjects with schizophrenia will eventually be admitted to hospital and therefore little bias is introduced by restricting research to hospitalized subjects. Using the Lothian Psychiatric Case Register, 66 subjects were identified who had been diagnosed in Edinburgh as suffering from schizophrenia between 1978 and 1989 but had no history of hospital admission by December 1991. This represented an adjusted average of 6.7% of the estimated annual rate of first diagnosis of schizophrenia: the proportion of such patients did not change over the period. Using a case-control design, the index cases were compared with a control group of schizophrenic patients who had been admitted to hospital within 3 months of diagnosis. At the time of diagnosis, the cases were generally less disturbed with lower levels of violent behaviour and less evidence of neglect or hallucinations. They had a longer duration of illness prior to diagnosis, were more often diagnosed by a consultant and unemployed. In a follow up study of the index cases, 59 (89%) were traced, of whom 6 (10%) were deceased. The outcome of the illness was heterogeneous although the course was often chronic. The general practitioner provided most of the care they received. The small proportion of such patients suggests that their exclusion from most published series does not seriously bias our picture of the natural history of schizophrenia. Moreover, as there was no increase in the proportion over the period, first admission rates for schizophrenia in Scotland are a reasonable approximation to incidence rates. PMID- 7480465 TI - Acute stress response: a comparison of head injured and non-head injured patients. AB - Acute stress responses were investigated in head injured (N = 38) and non-head injured (N = 38) motor vehicle accident victims immediately following their trauma. Results indicated that amnesia of the traumatic event was associated with less acute post-traumatic stress, intrusive symptoms, perceived injury, and fear of future risk. A proportion of head injured patients reported intrusive and avoidance symptoms despite being amnesic of their trauma. Findings are discussed in terms of the differential post-traumatic adjustment of head injured and non head injured traumatized patients. PMID- 7480466 TI - The relationship between suggestibility and anxiety among suspects detained at police stations. AB - The present study investigated the relationship between anxiety, as measured by the State-Trait Inventory (Spielberger, 1983) and interrogative suggestibility, as measured by the Gudjonsson Suggestibility Scale (GSS 2, Gudjonsson, 1987). One hundred and sixty-one suspects detained at two English police stations for questioning were assessed prior to being interviewed by the police. Unexpectedly, trait anxiety correlated more consistently with suggestibility than did state anxiety. The correlations were consistently higher among the Caucasian than the Afro-Caribbean subjects and the Afro-Caribbean subjects were significantly more suggestible than the Caucasian subjects even after their GSS 2 memory and IQ scores had been controlled for. The main practical implications of the findings are that interrogative suggestibility cannot be easily evaluated from the person's self-reported anxiety and the situation in which people are assessed may influence the relationship between these psychological variables. PMID- 7480467 TI - On the accuracy of personality judgment: a realistic approach. AB - The "accuracy paradigm" for the study of personality judgment provides an important, new complement to the "error paradigm" that dominated this area of research for almost 2 decades. The present article introduces a specific approach within the accuracy paradigm called the Realistic Accuracy Model (RAM). RAM begins with the assumption that personality traits are real attributes of individuals. This assumption entails the use of a broad array of criteria for the evaluation of personality judgment and leads to a model that describes accuracy as a function of the availability, detection, and utilization of relevant behavioral cues. RAM provides a common explanation for basic moderators of accuracy, sheds light on how these moderators interact, and outlines a research agenda that includes the reintegration of the study of error with the study of accuracy. PMID- 7480468 TI - Aesthetic properties of pictorial perception. AB - The present study assessed the dynamic person-object relationship in pictorial perception as associated with a high-level experience like the aesthetic. A series of experiments was designed to gain insight into the complexities of self world interactions involving the perceptual experience, which is the basis of artistic communication. Participants were asked to produce line drawings, which were then ranked by a larger group of participants as to whether they conveyed expressive contents. This resulted in considerable agreement among the judges. In addition, the factor structure of the semantic differential responses was determined. The results indicated that considerable semantic space existed between them. A theory of a linkage between affective and cognitive processes in perceptual experience is proposed. PMID- 7480469 TI - Contributions to psychohistory: XXIII. Hemingway "analyzed". AB - This paper is a review of major published works from psychoanalysts, psychiatrists, psychologists, academic scholars, and experts who have presented an "analysis" of the personality of Ernest Hemingway. Points of agreement and contrasting points of contention are discussed. PMID- 7480470 TI - A cautionary note about the cross-national and clinical validity of cut-off points for the Maslach Burnout Inventory. AB - In the present study, burnout scores of three samples, as measured with the Maslach Burnout Inventory, were compared: (1) the normative American sample from the test-manual (N = 10,067), (2) the normative Dutch sample (N = 3,892), and (3) a Dutch outpatient sample (N = 142). Generally, the highest burnout scores were found for the outpatient sample, followed by the American and Dutch normative samples, respectively. Slightly different patterns were noted for each of the three components. Probably sampling bias, i.e., the healthy worker effect, or cultural value patterns, i.e., femininity versus masculinity, might be responsible for the results. It is concluded that extreme caution is required when cut-off points are used to classify individuals by burnout scores; only nation-specific and clinically derived cut-off points should be employed. PMID- 7480473 TI - A new laboratory method for altering positive affect. AB - This paper reports the development with 95 undergraduates of a new method for altering Positive Affect in the laboratory. The method consists of assigning persons to complete a boring task for a specific amount of time and shortly after beginning the task, informing them that the assigned time period has either been increased or reduced. The advantages of this method are that it is effective, relatively free of demand characteristics, ethical, fast to administer, and parallels "real world" experiences. In addition, this procedure seems to circumvent limitations of other mood-induction procedures. PMID- 7480471 TI - Psychological adjustment to hospitalization: factorial structure, antecedents, and outcome. AB - Using various methods of observation, the psychological status of 211 patients of both sexes was assessed after the surgery or treatment justifying their hospitalization. Intercorrelations of scores on 40 variables obtained to describe the state of the patients were submitted to a principal components analysis. The latter yielded a factor of maladjustment to hospitalization associated with various psychosociological antecedents of the patients and a short-term unfavorable development of their physical health. This dimension seems to correspond not just with personal characteristics but to involve the interaction of medical staff members and patients in a manner detrimental to patients' recovery. PMID- 7480472 TI - Dissociation and somatization in adolescent inpatients with and without history of abuse. AB - 33 adolescents were tested for signs of dissociation and somatization as these processes relate to histories of physical and sexual abuse, a characteristic of over half of patients admitted for acute psychiatric disorder. Analysis suggested that the 19 adolescents having histories of abuse were more likely to score higher on dissociation and somatization. PMID- 7480475 TI - Associations of stressful life events and marital quality. AB - This is an exploration of whether stressful life events were related to 200 couples' marital quality after four or five years of marriage. Implications for measuring influence of stressful life events on marital quality are discussed. PMID- 7480474 TI - Prejudice toward persons living with a fatal illness. AB - Vignettes depicting a person living with a fictitious fatal illness were presented to 222 undergraduates. Manipulated variables in a completely randomized 3 x 2 x 2 factorial design were method of transmission of the illness (genetic/contagious/infectious), the population likely to become ill (anyone/primarily marginal persons), and the amount of suffering (little/much). Subjects reported on perceptions of ease of transmission of the illness, danger of contact with the ill person, blame, support of sanction, and social distance. Multivariate analysis of covariance showed that the manipulated variables affected the dependent variables considered together. Greatest prejudice was shown toward persons suffering greatly from an infectious illness that affected primarily marginal persons. Results are discussed relative to HIV. PMID- 7480477 TI - Childhood trauma and personality disorder in 43 women with panic disorder. AB - Among 43 women with panic disorder (38 with agoraphobia), 23 reported histories of childhood sexual abuse associated with high incidence of other early adverse experiences. The finding that subjects reporting childhood sexual and physical abuse were more likely to receive an Axis II diagnosis than those who did not confirms prior work. PMID- 7480476 TI - Different HIV risk profiles in samples of college students and homeless persons. AB - A cross-sectional survey examined demographic characteristics, self-reported sexual behaviors, and knowledge of AIDS in samples of 106 homeless persons and 260 college students. As expected, the two samples differed with respect to age, gender, race, and education. Respondents in both samples possessed moderate knowledge of HIV infection and AIDS and reported they considered their personal risk of HIV infection low. Both samples acknowledged frequent use of alcohol with sexual activity, active and passive oral sex, and lax use of condoms. A discriminant analysis indicated that a greater percentage of college students were sexually active than of the homeless sample and that the homeless group had a higher proportion of individuals who were either homosexual or bisexual, who had sexual contact with multiple partners or who had visited a prostitute, who had sexually transmitted diseases, and who had injected drugs. Although the risk profiles differed, each group reported high-risk behaviors and perceptions of low personal risk of HIV infection. PMID- 7480478 TI - Further validation of the Self-Defeating Personality Scale. AB - In a sample of 51 women and 35 men those who scored higher on the Self-defeating Personality Scale reported having "dropped out" of useful activities more often and scored as less assertive on the Assertiveness Self-report Inventory. Women were more likely than men to report having experienced at least one self defeating relationship. Attempts to replicate gender differences reported earlier were partially successful. The results provide additional validation for the Self defeating Personality Scale. PMID- 7480479 TI - Personality traits of women with breast cancer: before and after diagnosis. AB - The aim of this study was to investigate the extent to which the diagnosis of breast cancer influences the self-assessment of personality traits by patients. In 1989 and 1990, all female inhabitants of Nijmegen aged 43 years and over were invited to participate in a population screening for breast cancer and were asked to complete a personality questionnaire. During 1989-1992, 25 women who were diagnosed as having breast cancer were psychologically assessed for a second time 1 1/2 years after diagnosis. A control group of 822 healthy women also completed the questionnaire twice. A statistically significant decrease in scores on three scales (Rationality, Emotional Expression-out, and Emotional-Control) was reported by the patient group compared to the control group. PMID- 7480480 TI - Judgments of substance use and creativity in 'ordinary' people's everyday lifestyles. AB - To investigate the relationship between substance use and creativity, four pairs of raters independently assessed the reports of over 600 respondents from the Mass-Observation (M-O) Archive. The M-O directive addressed, their use of 'stimulants and relaxants.' Correlations between 'creativity' ratings and 'substance use' ratings were low but significantly positive (around .275). The observed links are discussed in relation to laboratory-based and other studies of substance use and creativity. PMID- 7480481 TI - Children's occupational sex-role stereotyping in 1994. AB - To update the basis of the explanation of how elementary children evaluate the capability of males versus females to perform occupations which are dominated by males, females, or are gender balanced (neutral) in the workforce 180 girls and 200 boys in Grades 1, 2, and 3 completed a questionnaire indicating which of 39 occupations were mostly done by women, by either, or by men on a 5-point scale. The most and least stereotyped occupations were very like gender-occupational data from 1972 and 1983 and reflected the actual sex-typing in the labor force. Sex and grade differences were noted for male occupations but not for female occupations. Some differences from 1977 data of Garrett, et al. were noted. The neutral status of some occupations appeared to assume a male worker. PMID- 7480482 TI - Self-defeating personality and depression: a closer look. AB - 55 men and 55 women were given Schill's Self-defeating Personality Scale, a measure of Beck's cognitive triad, and a measure of the personality dimensions of sociotropy and autonomy. The more characteristics of self-defeating personality endorsed, the more negative were subjects' cognitions about themselves, the world, and the future. Self-defeating men and women scored higher on sociotropy (social dependence), suggesting threats to relationships may be an important source of depression for them. Self-defeating women also scored higher on the autonomy subscales, suggesting threats to their independence may be a source of depression as well. Implications for treatment were discussed. PMID- 7480483 TI - Time series analysis of improved quality of life in Canada: social change, collective consciousness, and the TM-Sidhi program. AB - Two replication studies test in Canada a field theory of the effect of consciousness on social change. The exogenous variable is the number of participants in the largest North American group practice of the Transcendental Meditation and TM-Sidhi program, in Iowa. The first study indicated a significant reduction in violent deaths (homicide, suicide, and motor vehicle fatalities), using both time series intervention analysis and transfer function analysis methods, in weeks following change in the exogenous variable during the period 1983 to 1985. The second study, using time series intervention analysis, gave during and after intervention periods a significant improvement in quality of life on an index composed of the behavioral variables available on a monthly basis for Canada from 1972 to 1986-homicide, suicide, motor vehicle fatalities, cigarette consumption, and workers' days lost due to strikes. Implications of the findings for theory and social policy are noted briefly. PMID- 7480484 TI - Treatment of sexual offenders in an outpatient community-based program. AB - The components of a court-mandated outpatient treatment program in Florida for adult sex offenders are discussed. Selection and treatment factors are presented, in addition to the success rate, in terms of recidivism for program participants (N = 202) over a 5-year time span. This study is an extension of the 1989 work of Bingham and Piotrowski. PMID- 7480485 TI - Decrease violence in a day program: community-based endeavors. AB - Aggressive behaviors of two men with mental retardation were investigated during sheltered workshop and community employment. The first year of community work showed a monthly average reduction in aggressive behaviors of 93%. Changing to a typical work setting with regular expectations led to this decrease. PMID- 7480487 TI - An exercise for increasing students' understanding of life-span developmental psychology. AB - During the first week of a class in developmental psychology, students listed 5 to 7 characteristics they believed represent life-span developmental characteristics for the categories of infants, young children, adolescents, middle-aged adults, and older adults. Students retained their lists which were referred to during discussion throughout the course. Students reported this activity increased their understanding of topics central to life-span developmental psychology. PMID- 7480488 TI - Comparison of undergraduates with older adults on love styles and life satisfaction. AB - In a previous study, sex differences were noted in the correlation of 215 undergraduates' ratings on Lee's love styles and their ratings on their satisfaction with life, work, and personal relationships. For example, game playing love was positively related to life satisfaction for the 42 undergraduate men but was negatively related to life satisfaction for the 98 undergraduate women. In the current study, 215 adults over the age of 29 years were surveyed to examine whether the previous differences between the sexes held up in an older sample. Values for the older sample of 215 were similar to those for the 98 undergraduate women but not for the 42 undergraduate men. For example, game playing love was negatively related to life satisfaction for both the 130 older women and the 85 older men. PMID- 7480486 TI - Effectiveness of quality of life therapy for depression. AB - A bibliotherapy outcome study was conducted to assess the efficacy of Quality of Life. Therapy for depression. Sixteen clinically depressed community volunteers who showed an aptitude for and interest in bibliotherapy and were not suffering from other disorders met weekly to discuss a manual on Quality of Life Therapy. All subjects who completed treatment were reclassified as nondepressed and showed significant increases in quality of life and self-efficacy at the end of treatment. All but one subject maintained these improvements at a follow-up assessment. The potential for increasing the improvement rate of a treatment by matching patients to treatment modalities that fit their aptitudes, skills, and interests is discussed. PMID- 7480489 TI - Perceptions of disability by students, the formerly obese, and the general community. AB - There has been recurring interest over nearly three decades in preferences for different disabilities. In the present study, the perceptions of eight disabilities were compared across three groups, 42 occupational therapy students, 20 formerly obese, and 166 of the general community. A disability ranking task was adapted from a forced-choice task used by other researchers. Consistent with previous results, blindness was ranked as most difficult to live with. Contrary to prediction, there was no significant difference between the formerly obese subsample and the general community in their ranking of obesity; however, these groups differed in their ranking of diabetes. There were minor differences between the students and other groups. Across the entire sample, the disability of 'needing a hearing aid' was ranked as easiest to live with. Implications of the results are discussed, and suggestions made for further research. PMID- 7480490 TI - Split-half reliability of the Work Addiction Risk Test: development of a measure of workaholism. PMID- 7480492 TI - Faking and its detection on the Holden Psychological Screening Inventory. AB - This paper describes an index for the detection of fakers on the Holden Psychological Screening Inventory. Initially, the index is constructed and validated using an experimental paradigm. Subsequently, cut-off scores are developed based on a consideration of distributions of the index scores for the experimental groups (n = 101), for general psychiatric patients (n = 64), and for the general adult normative population (n = 564). PMID- 7480491 TI - Typological description of the chronic low back-pain syndrome using the Million Behavioral Health Inventory. AB - The Million Behavioral Health Inventory was administered to 67 individuals with chronic low back pain who participated in a rehabilitation program on pain and stress. Base-rate scores from the 20 scales, submitted to cluster analysis using Ward's minimum variance method, gave four subtypes. Discriminant analysis accurately classified 98.51% of the sample. The discriminant functions were interpreted and labeled as I: somatic distress and II: inhibition-dependency. The groups were then examined for differences and similarities with respect to those dimensions, and the groups were labeled I-introversive, II-denial/minimizers, III conformers, and IV-severe psychophysiological reaction. The types were next compared across medical, vocational, and demographic data. With the exception of sex of subject, no other data were significant by type. The results were discussed, and recommendations for research were made. PMID- 7480493 TI - Response to Sputa and Paulson's "Birth order and family size: influences on adolescents' achievement and related parenting behaviors". PMID- 7480494 TI - Development of the Relationship Beliefs Questionnaire. AB - The cognitive mediation model of human psychological functioning has received increased attention in examining the role of cognitive variables within distressed relationships. Development of the Relationship Beliefs Questionnaire, designed to assess adherence to relationship-specific irrational beliefs, is described. In 370 subjects, factor analysis yielded nine factors which made conceptual sense. On the total scale Cronbach alpha was .95. These results are described in terms of research and practice applications. PMID- 7480495 TI - Perceptions of childhood obesity of undergraduate students in physical education. AB - A sample (N = 200) of undergraduate students in physical education from 12 universities in a midwestern state was sent the 1990 Price questionnaire; 178 responded (89%). 96% of the respondents indicated that normal weight is very important in children, 88% agreed that physical education teachers should play major roles in treating childhood obesity. 92% believed their college courses prepared them to administer exercise programs to help children reduce weight, and 70% supported school-based weight-reduction strategies. Over-all, the students seemed to want to help eliminate childhood obesity and indicated they should become significantly involved in school programs designed to achieve this goal. PMID- 7480496 TI - Responses of noninterviewable long-term care patients before and after interinstitutional relocation. AB - This study investigated the health consequences of an interinstitutional relocation on a sample of 103 noninterviewable, cognitively impaired patients in long-term care. Relocated patients functioned as their own controls. Specifically, data were abstracted from the patients' medical records on 16 health indicators for the 6 months prior to and following relocation as well as for the same months in the year before. Multivariate analysis indicated patients in general exhibited good health pretransfer but worse health posttransfer. In addition, less impaired patients were somewhat more affected by relocation. PMID- 7480497 TI - Use of dreams in therapy: a survey of clinicians in private practice. AB - A literature review indicated a dearth of research on how dreams are used in therapeutic settings so a questionnaire was sent to a random sample of 500 members of the Florida Psychological Association to assess (a) the extent of dream use in therapy, (b) theoretical approaches applied to dream interpretation, and (c) basis for experience and expertise in dream work. Of the 500 potential clinicians, 228 returned survey forms for a response rate of 46%. Analysis indicated that 83% of the respondents used dream material at least occasionally in their practice and that Freudian and Gestalt approaches were most often used in dream interpretation. Interestingly, most respondents gained their 'experience' in dream work through self-study and continuing education workshops and seminars. PMID- 7480498 TI - Attitudes of Hong Kong medical students toward different modalities of treatment for cancer. AB - 140 medical undergraduates in a Hong Kong medical school rated the usefulness of different modalities of cancer treatment for curative, medically palliative, and psychologically palliative care, respectively. Factor analyses showed that respondents used a Chinese versus non-Chinese cultural framework in conceptualizing modalities of treatment outside the conventional Western medical model. The functions of unconventional modalities were mainly for psychologically palliative care according to the respondents, although they did not regard these modalities as adversive to curative and medically palliative treatments. Among the Western medical modalities, surgery was rated as more useful for curative treatment while chemotherapy and radiotherapy were rated as more useful for symptom control. The implications of the findings for cancer care and further research were discussed. PMID- 7480499 TI - Alexithymia and coping with stress in healthy persons: alexithymia as a personality trait is associated with low social support and poor responses to stress. AB - We examined the relationship between alexithymia and coping with stress among 179 Japanese healthy volunteers. Two correlational analyses indicated that persons scored as alexithymic were more likely to indicate lower social support and poorer responses to stress. Multiple regression analysis also indicated that these lowered coping responses were fully explained by alexithymia scores. These results suggest that alexithymia, as a personality trait, may help to explain these individuals' low social support and poor responses to stress. PMID- 7480502 TI - Economic factors and the rates of suicide in Germany between 1881 and 1989. AB - The potential consequence of economic stress most frequently cited in the literature of medical sociology is the increase in the rate of suicide, it probably being the most valid and reliable indicator of collective mental health. To assess the probability of such being the consequence of current economic realignment in the Federal Republic of Germany, we deemed it promising to evaluate the extent to and manner in which economic factors have to date affected the frequency of suicide in Germany. The current study analyzed the effects of four economic variables (growth of the economy, average real income, unemployment and frequency of bankruptcy) on the rates of suicide in Germany from 1881 to 1989. We set the commencement date of the period analyzed as early as possible to include long-term developments as well as the effects of different moderator variables. The annual fluctuations of all four variables, in conformity with our hypothesis, correlated both in the period preceding World War II as well as in the postwar period with those in the rates of suicide. The strongest correlations held for the rate of unemployment and for the frequency of bankruptcy in times of obvious social disintegration coupled with diminished state safeguards against unemployment. Our hypothesis that the effects of economic factors would more strongly influence the rates of suicide by men as opposed to women could not be corroborated. PMID- 7480501 TI - Reconstruction and augmentation patients' reaction to the media coverage of silicone gel-filled implants: anxiety evaluated. AB - Media coverage tends to focus on sensational aspects. Especially prominent are controversial items related to public health and well-being. This research examined the response by breast-reconstruction and breast-augmentation patients to extensive media coverage regarding silicone gel-filled implants. Results of our survey suggested that media coverage influenced these women. Common concerns expressed by respondents included autoimmune disease, capsular contracture, leakage, increased risk of cancer, and anxiety. All respondents used consultation with their physicians as a primary means of coping with anxiety. Anxiety over media coverage led to some hesitancy concerning use of implants by surveyed patients. Patients who experienced difficulty with their reconstruction noted more anxiety associated with media coverage. PMID- 7480500 TI - Fear of weight gain: its correlates among school-aged adolescents. PMID- 7480503 TI - Self-criticism and dependency in relation to anger. PMID- 7480504 TI - A study of planning and mathematics instruction for students with learning disabilities. AB - The purpose of this study was to extend research in training the use of cognitive strategies or planning to mathematical computation for 4 students with specific learning disabilities. A cognitive education method utilized in previous research was duplicated. It was expected that students would find the instruction differentially effective based upon their initial scores on a measure of planning. Using the Planning, Attention, Simultaneous, Successive model as a base, a cognitive instruction which facilitated planning was provided to two students with low scores on planning, obtained using an experimental version of the Das-Naglieri Cognitive Assessment System, and two students with average planning scores. All students completed three sessions of baseline and seven sessions of cognitive instruction in addition and multiplication. During the cognitive instruction phase, 5-min. sessions of self-reflection and verbalization of strategies about the mathematics problems were conducted after each initial 10 min. session of mathematics. Scores on addition problems showed that all students improved. On multiplication, however, 2 students with low planning scores improved considerably but not 2 with higher planning scores. Implications are provided. PMID- 7480505 TI - Persistence of SCII scores and CPI scores of students entering a baccalaureate nursing program during a seven-year period. Strong-Campbell Interest Inventory. PMID- 7480506 TI - Loneliness, duration of loneliness, and reported stress symptomatology. AB - The correlations between intensity of loneliness, duration of loneliness, and reported psychological and somatic stress symptoms were examined in 195 undergraduate students (100 women and 95 men). Analysis indicated that intensity and duration of loneliness were significantly correlated and both were also significantly positively correlated with subjects' reported psychological and somatic stress symptoms. In a forced-entry multiple regression equation predicting over-all stress symptomatology, only duration of loneliness emerged as a significant predictor, while intensity of loneliness did not add significant predictability. For college students, loneliness may be related to a wide array of reported psychological and somatic stress symptoms. It is also suggested that the duration of a lonely experience is perhaps a better predictor for symptoms of stress than intensity of loneliness. PMID- 7480507 TI - Abortion laws and infanticide. PMID- 7480508 TI - Perceived consequences to the predictor: a variable in the release of psychiatric patients. AB - The present study examined factors hypothesized to influence mental health professionals' perceptions of dangerousness, predictions of violence, and decisions on patients' release. 120 mental health professionals employed in state mental hospitals were each given one of 12 patient profiles. The independent variables, manipulated within vignettes, were (a) violence history, (b) paranoid schizophrenia versus nonparanoid schizophrenia, and (c) perceived consequences in terms of liability and publicity. Type of schizophrenia did not affect ratings, but violence history of the predictee and perceived consequences to the predictor did significantly influence the ratings. Patients with actual violence histories were viewed by the subjects as having more potential for future violence, as being more globally dangerous, and as requiring a more secure placement than those with histories of threats of violence or no violence. Possible litigation following release led to a recommendation for more secure placement than did minimal legal consequences. Predictions of violence and decisions on hospital release were interpreted as dependent on both predictor and patient-related variables. PMID- 7480509 TI - Is health locus of control related to health education activity. AB - Analysis of responses of 1922 teachers indicated that Multidimensional Health Locus of Control scale was a suitable instrument for assessing their health beliefs. The results suggest that health belief is not linked to whether teachers teach about cancer and, by implication, health generally. PMID- 7480510 TI - Microgenesis of the length perception of paired lines. AB - The changes in perception associated with exposure time are probably continuous in the microgenetic process, as is demonstrated by such phenomena as intensity change in illusions, adaptation and normalization, practice effect, aftereffects, etc. The simultaneous contrast and assimilation in the perception of length of paired lines were measured by controlling the exposure duration or repetition. In the tachistoscopic experiment, the greatest contrast appeared at an early stage of microgenesis. In the repeated presentation experiment, the perception changed from contrast to assimilation on the expanded stage of microgenesis. These changes in perception are assumed to be the results of the differentiation of set that progresses in the microgenetic process. In the fixed-set experiment, the differentiation of set was reflected in the intensity of the aftereffect. The changes in the aftereffect and the transformation into the assimilation coincided with each other when the exposure time was varied. An expansion of the conception of perceptual microgenesis to a wider range of perceptual variability is accordingly proposed. PMID- 7480511 TI - The association of assimilation and an increase in visibility in perceptual grouping. AB - Subjects performed a series of forced-choice discriminations to determine whether both group-assimilation and group-visibility associations could be obtained from nearly identical strong and weak group patterns. The discrimination between the context+target square and the context [symbol: see text] was better than between the target- and background, as was the case for --, whose context and target components were its left and right halves, but not for [symbol: see text]. Square and -- produced a better performance when their lines (halves) were the same in color, and a poorer performance when their lines were different in color, but [symbol: see text] produced the reverse. Likewise, only square and -- produced a better performance when closed, and a poorer performance when open. These context+target etc., same-different, and closure results argue that square and -- produced a greater increase in visibility of their component --, more assimilation among their parts, and a stronger group than did [symbol: see text]. This evidence of a group-assimilation-visibility association cannot be attributed to the fortuitous occurrence of an increase in visibility with one object, assimilation with a second, and closure with a third, unlike previous evidence. This association cannot be explained by feature-based theories. Therefore, a superordinate unit is the cause of this association. PMID- 7480512 TI - The effect of a masked stimulus on the response to the masking stimulus. AB - Four experiments are reported in which the subjects had to respond to a target that masked a preceding prime via metacontrast masking. In one part of Experiment 1, the subjects discriminated the target's shape (square or diamond) by a motor choice reaction, and in another part they had to respond with a simple reaction. The prime was neutral (circular) with respect to the target's shape. The data showed a facilitation effect. In both tasks the reaction time was reduced by the masked prime. However, the reduction was more pronounced with simple reaction than with choice reaction. In the other experiments, additional primes were used with the same angular shapes as the targets. In Experiments 2 and 3, after discriminating the target's shape by a choice reaction, the subjects had to judge the prime's shape in a signal-detection task. While neither the d' value for discriminating the angular primes from the circular ones (Exp. 2) nor the d' value for distinguishing between the angular primes (Exp. 3) was different from zero, the choice-reaction data showed a congruency effect. With a congruent prime (i.e., a prime that had the same shape as the target), the reaction times were reduced. With an incongruent prime, the reaction times grew. In Experiment 4 the errors were investigated. The facilitation effect was present in the RT, but not in the number of errors, whereas the congruency effect was present in the number, but not in the RT of errors. While the facilitation effect can be attributed either to an unspecific activation by the masked prime or to an influence of the prime on attentional processes, the congruency effect can be explained by the assumption that the masked prime directly activates the specific response, which corresponds to the prime's shape. PMID- 7480513 TI - Acute effect of altered tryptophan levels and alcohol on aggression in normal human males. AB - Normal males received amino acid mixtures designed to raise or lower tryptophan availability, and thus to raise or lower brain serotonin synthesis. They also received alcoholic or non-alcoholic drinks. The subjects were tested in the Taylor Competitive Reaction Time Task in which they competed against a (non existent) partner in a reaction time task. The magnitude of electric shocks that the subjects were willing to give to their bogus partner was used as a measure of aggression. Lowered tryptophan levels and ingestion of alcohol were associated with increased aggression. Our data support the idea that low serotonin levels may be involved in the etiology of aggression. They suggest that subjects with low brain serotonin levels may be particularly susceptible to alcohol-induced violence. PMID- 7480514 TI - Characterization of topographic EEG changes when smoking a cigarette. AB - The acute effects of cigarette smoking on the human electroencephalogram (EEG) were investigated by the topographic mapping technique. Twenty-six subjects participated in this study, which involved sham smoking and real smoking of preferred cigarettes. Effects of smoking were analyzed by statistical and multivariate analysis. Analysis of variance and t-test results showed a significant decrease in the theta and alpha 1 bands but a significant increase in the alpha 2, beta 1, and beta 2 bands. Factor analysis and cluster analysis showed that there were two or three independent regions on the scalp that indicate the effects of smoking on topographic EEG. A hypothesis was formed that smoking has different effects on human EEG profiles for different brain regions and that there are individual variations in the EEG responses to smoking. PMID- 7480515 TI - ABT-418: discriminative stimulus properties and effect on ventral tegmental cell activity. AB - Previous studies have established that ABT-418 [(S)-3-methyl-5-(1 methyl-2 pyrrolidinyl)isoxazole hydrochloride] is a novel neuronal nicotinic acetylcholine receptor (nAChR) ligand with cognitive enhancing and anxiolytic-like activity 3- to 10-fold more potent than (-)-nicotine in rodents. A series of experiments was conducted to determine the discriminative stimulus properties of ABT-418 in comparison with (-)-nicotine, and to determine the relative potencies of these compounds on ventral tegmental area (VTA) neurons. While rats were able to discriminate (-)-nicotine 1.9 mumol/kg in 39 days, they were not able to discriminate 1.9 or 6.2 mumol/kg ABT-418 from a saline solution during 50 days of training. In rats trained to discriminate 1.9 mumol/kg (-)-nicotine, a reduced generalization was induced by ABT-418 at 1.9 and 6.2 mumol/kg, an effect completely blocked by the cholinergic channel blocker mecamylamine (15 mumol/kg, IP). However, in extensively trained rats, intraperitoneal or subcutaneous injections of ABT-418 induced 78-82% generalization at the 6.2 mumol/kg dose. The predominant metabolites of (-)-nicotine and ABT-418 (continine and A-87770, respectively) were devoid of any effect in nicotine-trained rats. The reduced potency of ABT-418 in nicotine-trained rats is consistent with the electrophysiological findings showing that ABT-418 is 3-fold less potent than (-) nicotine in activating dopamine-containing neurons in the VTA area. PMID- 7480516 TI - Rate of binding of various inhibitors at the dopamine transporter in vivo. AB - The rate of entry of drugs into brain is thought to be a factor in their abuse liability. In this investigation, we have examined the rate of entry and binding at dopamine transporters in mouse striatum for a variety of dopamine transporter inhibitors. The method utilized was based on measuring the displacement of 3H-WIN 35,428 from striatal dopamine transporter sites in vivo at different times. Eleven cocaine analogs (RTI-31, RTI-32, RTI-51, RTI-55, RTI-113, RTI-114, RTI 117, RTI-120, RTI-121, WIN 35,065-2, and WIN 35,428) as well as other dopamine uptake site blockers (bupropion, nomifensine, and methylphenidate) were compared with (-) cocaine for their rates of displacement of 3H-WIN 35,428 binding in vivo. The drugs that displayed the fastest occupancy rates were bupropion, (-) cocaine, nomifensine, and methylphenidate. RTI-51, RTI-121, RTI-114, RTI-117, RTI 120, RTI-32, RTI-55, and RTI-113, showed intermediate rates, whereas RTI-31, WIN 35,065-2, and WIN 35,428 exhibited the slowest rates of displacement. While many of the cocaine analogs have proven to be behaviorally and pharmacologically more potent than (-) cocaine, their rates of entry and binding site occupancy were slower than that for (-) cocaine. Earliest times of transporter occupancy by the different drugs were correlated (although weakly) with their degree of lipophilicity (r = 0.59; P < 0.02). Kinetic effects and metabolism of the compounds could complicate the interpretations of these data.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7480517 TI - Response to smoking as a function of prior smoking amounts. AB - Smoking patterns are changing to accommodate new environmental smoking restrictions. The current study explored the effects of prior smoking amounts on the behavioral, physiological, and subjective effects of smoking a single cigarette. Ten smokers (six females, four males) each participated in four laboratory sessions of 6 h duration. During each session, they smoked 0, 2, 5, or 11 cigarettes which were evenly spaced throughout the 6-h period at intervals ranging from 30 min (11 cigarettes) to 120 min (2 cigarettes), with condition order determined by a Latin square. At the end of each session, all smoked a final cigarette. Response to the last cigarette of the 6-h session was influenced by pretreatment smoking amounts. Number of puffs drawn from the cigarette (15.0, 14.1, 13.3, and 10.1) was inversely related to prior smoking density, as was heart rate increase (5.8, 4.0, 2.4, and 1.3 bpm). Three physical symptoms of smoking, dizzy, lightheaded and tingling, were significantly greater when preceded by 6 h of no smoking than when preceded by smoking at 30-min intervals. However, these symptoms were rated as mild (19-27 on a 100-point scale) even when no cigarettes had been smoked. These results suggest some dissipation of acute tolerance after 6 h of smoking abstinence. Liking and satisfaction ratings were inversely related to the pretreatment density of smoking. Liking scores for the last cigarette were 85, 71, 68 and 48 (on a 100-point analog scale) when 0, 2, 5 and 11 cigarettes had been smoked. Overall, the study showed that physiological and subjective effects of smoking are modulated by recent histories of smoking.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7480518 TI - Velnacrine maleate improves delayed matching performance by aged monkeys. AB - Velnacrine maleate is a novel, orally active acetylcholinesterase inhibitor of the acridine class with a longer duration of action than physostigmine. Velnacrine has shown efficacy in the treatment of Alzheimer's disease and in improving both normal and experimentally impaired mnemonic function in animals and humans. To characterize this action further, the present study evaluated velnacrine for its ability to ameliorate the decline in short-term memory associated with aging in non-human primates at two time points after velnacrine administration: (1) 30 min and (2) 24 h. Initially, doses of 1, 2, 4, and 6 mg/kg, PO (free base corrected) were administered once to each of six aged (25-40 years), memory-impaired macaques that had been trained to perform a delayed matching-to-sample (DMTS) paradigm. The dose associated with the greatest improvement in session performance was administered three more times to the same individual. Four of the six monkeys showed improved DMTS performance during the repeated best dose phase (phase 2). Almost all of the improvement occurred during long-delay trials. Compared to placebo trials, velnacrine induced a significant improvement of long delay DMTS (58.0-66.7%, 13.4% of the placebo value). Long delay DMTS remained significantly improved during the test session conducted 24 h following velnacrine administration. Pharmacokinetic analysis following administration of 4 or 6 mg/kg velnacrine to three aged monkeys revealed peak plasma concentrations ranging from 27 to 166 ng/ml, 30-60 min after dosing. Six hours after dosing velnacrine plasma levels decreased to 5.1-11.8 ng/ml; and 24 h after dosing velnacrine plasma levels were less than the limit of quantitation (5 ng/ml).(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7480519 TI - State-dependent effects of atypical benzodiazepine-receptor agonists. AB - The state-dependent effect of the BZ-receptor agonist diazepam (1.25-10 mg/kg), the partial agonist FG 8205 (0.5-4.0 mg/kg) and the BZ1-receptor agonist zolpidem (0.25-2 mg/kg) were investigated in rats. During daily sessions, animals were trained to acquire FR10 lever pressing for food reinforcement whilst under the influence of the agonists, using an operant technique. Forty-eight hours after the final training session under drug, their performance of the FR10 was evaluated during a test session, carried out following vehicle administration only. Neither diazepam, nor FG 8205 impaired acquisition of the task. In the group treated with 2 mg/kg zolpidem, six out of eight rats failed to learn within 20 sessions, but the smaller doses were without effect on acquisition. When drug treatment was withdrawn, there was evidence that all three of the agonists tested produced state-dependency. This was apparent in the form of longer latencies to obtain reinforcement and decreased lever pressing rates. The significance of these findings are discussed in the context of the relationship between the state dependent effects of BZ-receptor agonists and their other properties, and the receptor subtypes which might underly these effects. PMID- 7480521 TI - Sensitization to apomorphine in pigeons: unaffected by latent inhibition but still due to classical conditioning. AB - When administered apomorphine, pigeons exhibit protracted bouts of pecking behavior. This response is subject to sensitization, as it initially increases with repeated drug injections. The hypothesis is examined that the sensitization is due to a Pavlovian conditioning of the drug-induced pecking to the environment in which it first takes effect. In a first experiment, we attempted to suppress this conditioning by extensively pre-exposing the birds to the test environment and saline injections (latent inhibition procedure). As the experiment yielded undiminished sensitization, it cast doubt on the conditioning hypothesis. However, while inhibitory pretraining also proved ineffective in a second experiment, a shortening of response latencies specific to the environment in which the animals had first experienced the apomorphine effect supported the conditioning hypothesis. It is suggested that the absence of latent inhibition may be due to the interference of a context-dependent conditioning effect. A third experiment that examined the hypothesis that the reinforcing properties of apomorphine might be attributable to its well known anorectic properties. The results provided some support for this notion. At the same time, they also confirmed that apomorphine-induced pecking conditions reliably to environmental cues. These cues are then by themselves capable of provoking conditioned pecking. PMID- 7480520 TI - Effects of post-trial administration of nicotine on human memory: evaluating the conditions for improving memory. AB - In the studies reported here, we investigated the effects of nicotine on memory for unrelated word lists. Nicotine was delivered through cigarette smoking, and memory performance was assessed using both intentional and incidental recall tasks, and employing an additional, indirect measure of memory. We report the results of four experiments in which we manipulated 1) the intake of nicotine using nicotine-containing and nicotine-free cigarettes, 2) the associative aspects of the word-sets, by unrelated words and category words and by instructing subjects to use an associative mnemonic strategy, 3) the opportunity for rehearsal between the presentation and recall, and 4) the time of nicotine administration, post- or pre-trial. We found a positive effect of post-trial nicotine on memory in the incidental recall task, as indicated by enhanced repetition priming, but no effect of nicotine on either immediate recall or pronunciation times (experiments 1 and 2). In experiment 3 we examined the effects of post-trial nicotine using associative and unrelated word-lists, when volunteers were instructed to use an associative mnemonic strategy. We found no main effect of nicotine, but when volunteers were distracted from rehearsal, related words were recalled better than unrelated words. Experiment 4 was a positive control for the timing of nicotine administration within our experimental design, and this showed that pre-trial nicotine not only improved free recall but differentially enhanced the recall of category words over unrelated words.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7480522 TI - Primacy and recency effects in rhesus monkeys (Macaca mulatta) using a serial probe recognition task. I. Effects of diazepam. AB - In this study, we evaluated the effects of diazepam (0.2, 0.8, 1.6 and 3.2 mg/kg; IM) on the primacy and recency memory effects in four rhesus monkeys trained on a six-item serial probe recognition (SPR) task. Only the highest dose of diazepam (3.2 mg/kg) consistently affected the shape of the monkeys' serial position curves. Accuracy on the probe trials was disrupted for list items which occurred in the middle portion and recency memory component of the serial position curve, without affecting the primacy component. Diazepam, however, also produced several nonspecific effects on SPR performance. Both the 1.6 and 3.2 mg/kg diazepam doses disrupted accuracy on the non-matching probe trials, and the 3.2 mg/kg dose caused an increase in response latencies which were unrelated to any one component of the serial position curve. This is the first demonstration in nonhuman primates showing that the primacy and recency memory effects can be differentiated on the basis of diazepam dose. PMID- 7480523 TI - The olfactory tubercle as a site of action of neuroleptics with an atypical profile in the paw test: effect of risperidone, prothipendyl, ORG 5222, sertindole and olanzapine. AB - The paw test was used to detect the preclinical profile (classical versus atypical) of five putative, atypical neuroleptics, namely olanzapine, sertindole, risperidone, prothipendyl and ORG 5222. In the paw test classical neuroleptics increase the hindlimb reaction time (HRT), a parameter with predictive validity for antipsychotic efficacy, at doses comparable to those necessary for increasing forelimb reaction time (FRT), a parameter with predictive validity for extrapyramidal side-effects, whereas atypical neuroleptics increase HRT at doses that are much smaller than those increasing FRT. All tested compounds showed the profile of atypical neuroleptics in the paw test. Using the FRT/HRT ratio of minimum effective doses as overall predictor of a favourable ratio of extrapyramidal and therapeutic effects of these drugs, the following order was found: olanzapine (20) > sertindole = risperidone = prothipendyl (10) > ORG 5222 (3). The ability of compounds to attenuate locomotor activity elicited either from the olfactory tubercle (10 micrograms dopamine: OT test) or from the nucleus accumbens (1 microgram ergometrine: ACC test) was used to establish whether the compounds preferentially act in one of these structures. Previous research has shown that classical neuroleptics are far less potent in the OT test than in the ACC test, whereas atypical neuroleptics are far more potent in the OT test than in the ACC test. All five agents preferentially acted in the olfactory tubercle. The order of potency in the olfactory tubercle was as follows: sertindole > ORG 5222 > risperidone > olanzapine > prothipendyl. It is concluded that risperidone, prothipendyl, ORG 5222, sertindole and olanzapine not only show the profile of atypical neuroleptics in the paw test, but also preferentially act in the olfactory tubercle, but not in the nucleus accumbens, viz. two features that they share with the atypical neuroleptics clozapine and thioridazine and with the putative, atypical neuroleptics raclopride and remoxipride. PMID- 7480524 TI - Onset and time course of antidepressant action: psychopharmacological implications of a controlled trial of electroconvulsive therapy. AB - Onset and time course of antidepressant effect were examined in 47 patients with major depressive disorder who had been randomly assigned to twice weekly bilateral, brief pulse electroconvulsive therapy plus one simulated treatment per week (ECTx2) or to a three times weekly schedule of administration (ECTx3). Rapid improvement was observed in the ECTx3 group in whom the number of real ECTs to 30% reduction on the Hamilton Depression Scale (HAM-D) was 3.2 +/- 1.90, administered over 7.3 +/- 4.43 days and to 60% reduction, 5.9 +/- 3.09 real ECTs over 13.7 +/- 7.21 days. Among the responders in both groups combined, 24.3 +/- 29.58% of the overall improvement in HAM-D was contributed by the first real ECT, 60.9 +/- 28.13% by the first four real ECTs and 91.6 +/- 25.82% by the first eight. Although 85.3% of the responders had reached 60% HAM-D improvement after eight ECTs, a clinically significant minority (14.7%) responded later in the course (ECT 9-12). However, response was predictable on the basis of symptomatic improvement (30% on the HAM-D) by the sixth real ECT. Thirty-three out of 34 responders would have been correctly identified by this criterion and only 2 out of 13 non-responders mis-identified (P < 0.000001). Once achieved, the antidepressant effect was stable, without continuation pharmacotherapy, until 1 week after the last treatment and on lithium carbonate (Li) or Li plus clomipramine for a further 3 weeks. These findings confirm the clinical impression that ECT is a rapidly effective treatment for major depression with a shorter latency than generally reported for antidepressant drugs.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7480525 TI - Clinical trials of antidepressants: the hidden face: where locus of control appears to play a key role in depression outcome. AB - It remains difficult to determine in what measure improvements observed in clinical trials of anti-depressants may be attributable to the psychological predispositions of the subjects. The present article focuses on the effect of a psychological variable, the Health Locus of Control, which measures the extent of a subject's belief that he is in control over his own health. The hypothesis is that depressed subjects whose locus of control is internal, i.e. who perceive themselves to be in control, rather than external, i.e. control perceived as being in others or just chance, will improve more markedly and consistently on the Hamilton Depression Rating Scale, across a number of clinical trials. Forty nine depressive patients undergoing treatment with four different compounds were included, after a week's placebo run-in period, in a classical 42-day follow-up study comprising visits on days -7, 0, 10, 21; and 42. Interactions between the type of locus of control and the clinical course were investigated by MANOVA. Results show that with a classical design of clinical trials of antidepressants, locus of control plays a significant role if it is internal (P < 0.001) in consolidating the improvement process, and that this is true irrespective of type of antidepressant. The relationship between the concept of locus of control and placebo effect is discussed. PMID- 7480526 TI - Effects of ethanol on the processing of low probability stimuli: an ERP study. AB - The effect of a moderate dose of ethanol on the processing of low probability task-relevant and task-irrelevant stimuli was investigated using event-related potentials (ERPs). Sixteen subjects received alcoholic and placebo beverages on alternate days. ERPs were recorded from 15 locations on the scalp. The subjects were asked to press a button upon detection of rare target stimuli embedded among frequent standard and rare "novel" stimuli. Ethanol 1) reduced the amplitude of P3 to novel stimuli, but not P3 to target stimuli, 2) did not affect the mismatch negativity, and 3) delayed P3 latency and reaction time independently. These results suggest that 1) the processing of rare task-irrelevant stimuli is more vulnerable to the effects of ethanol than is the processing of task-relevant stimuli, and 2) ethanol impacts stimulus evaluation time and response production stages of information processing. PMID- 7480528 TI - The mitochondrial toxin 3-nitropropionic acid induces vacuous chewing movements in rats. Implications for tardive dyskinesia? AB - Rats were chronically treated for 4 weeks with three doses (4, 8, 12 mg/kg per day) of the mitochondrial toxin 3-nitropropionic acid (3-NP). The behaviour was videotaped at intervals during and after treatment, and vacuous chewing movements (VCM), a putative analogue to tardive dyskinesia (TD), as well as the general behaviour were scored. During treatment, 3-NP dose-dependently increased VCM and dose-dependently reduced motor activity, which is similar to the behavioural effect of long-term neuroleptic treatment. The results may support the hypothesis that neuroleptic-induced impairment of energy metabolism, a potential excitotoxic mechanism, is involved in the development of VCM in rats and probably TD in humans. PMID- 7480529 TI - Concurrent self-administration of ethanol and an alternative nondrug reinforcer in monkeys: effects of income (session length) on demand for drug. AB - Eight rhesus monkeys (Macaca mulata) were trained to self-administer orally delivered ethanol (8%) and saccharin (0.03 or 0.3% wt/vol) or water under concurrent fixed-ratio (FR) schedules. The FR requirement for saccharin was fixed at 32, while the FR for ethanol was varied (4, 8, 16, 32, 64 and 128) in a non systematic order to assess demand for drug. Demand was defined as consumption plotted as a function of price (FR). Income was defined as the duration of access to available resources. Income was varied by allowing access to the concurrently available liquids 20, 60 or 180 min per day. Order of testing was counter balanced across monkeys. Saccharin deliveries were much higher than ethanol deliveries under the 180-min income condition; however, they were lower than ethanol deliveries when income was reduced to 20 min and the ethanol FR was 4, 8 or 16. Thus, when the price of drug was relatively low, consumption of drug exceeded that of the nondrug reinforcer, and that relationship was reversed as income decreased. Saccharin deliveries sustained a proportionally greater reduction due to decreased income compared to ethanol deliveries. As income decreased from 180 to 20 min, saccharin deliveries were reduced by an average of 79.1% (across ethanol FR conditions) while ethanol deliveries were reduced by an average of 41.2 and 40.8% when concurrent saccharin or water were available, respectively; thus, drug self-administration was more resistant to income changes than saccharin. The demand for ethanol was shifted downward in a parallel fashion as income decreased.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7480527 TI - Enhanced oral activity responses to intrastriatal SKF 38393 and m-CPP are attenuated by intrastriatal mianserin in neonatal 6-OHDA-lesioned rats. AB - Enhanced oral activity is induced in neonatal 6-hydroxydopamine- (6-OHDA-) lesioned rats by systemic administration of the dopamine (DA) D1 receptor agonist SKF 38393 and serotonin (5-HT) 5-HT2A,2C agonist m-chlorophenylpiperazine (m CPP). The DA D1 receptor antagonist SCH 23390 effectively attenuates the effect of SKF 38393 but not m-CPP. The 5-HT2 antagonist mianserin attenuates the effects of both m-CPP and SKF 38393, suggesting that DA agonist effects are mediated by 5 HT neurochemical systems. To test whether DA and 5-HT agonist effects and interactions might occur within the neostriatum, rats were implanted with permanent injection cannulae, with tips in the ventral striatum. One group of rats was lesioned at 3 days after birth with 6-OHDA HBr (100 micrograms salt form, in each lateral ventricle; desipramine HCl pretreatment, 20 mg/kg IP, base form, 1 h), while controls received the vehicle in place of 6-OHDA. Cannulae were implanted when rats weighed 200-250 g. During a 1-h observation session SKF 38393 (5 nmol per side) produced 74.3 +/- 19.2 oral movements in intact rats and 310.7 +/- 97.0 oral movements in 6-OHDA-lesioned rats. m-CPP (10 nmol per side) produced 72.6 +/- 15.1 and 274.5 +/- 65.0 oral movements in these respective groups. These responses were several-fold greater than the 25.3 +/- 7.3 and 41.8 +/- 9.5 oral movements in the same groups after saline (0.5 microliter per side) (P < 0.05). Mianserin (6 nmol per side) alone had no effect on oral activity but attenuated responses to both SKF 38393 and m-CPP in intact and 6-OHDA-lesioned rats.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7480531 TI - Treatment of depressive outpatients with lorazepam, alprazolam, amytriptyline and placebo. AB - This randomized double-blind study in 342 mildly to moderately depressive outpatients investigated the antidepressant effectiveness and speed of action of lorazepam, alprazolam and amitriptyline versus placebo. Six weeks of drug treatment were followed by a drug taper period, a control period with placebo and a control period without placebo, of 2 weeks duration each. Clinical improvement was assessed by rating scales (Clinical Global Impressions, Hamilton Rating Scales for Depression and Anxiety) and patient's self-ratings (Patient's Global Impressions, Self-rating Depression Scale and Visual Analogue Scale). At the end of week 6 all active drugs showed similar efficacy which was significantly superior to placebo. Compared to placebo, onset of efficacy was earlier on benzodiazepines than on amitriptyline . While tapering by decreasing the dosage, replacing drug with placebo and finally discontinuing placebo, clear withdrawal phenomena were not seen, but 20% of patients, equally distributed to all treatment groups, did not want to stop taking tablets after replacing drug with placebo. Drop-out rate during the treatment period was very low (9%). Significantly interfering adverse effects were seen in 27 patients, without predominance in one of the active drug groups. PMID- 7480530 TI - Fluctuations in nucleus accumbens dopamine concentration during intravenous cocaine self-administration in rats. AB - Fluctuations in extracellular dopamine and DOPAC levels in nucleus accumbens septi (NAS) were monitored in 1-min microdialysis samples taken from rats engaged in intravenous cocaine self-administration. For four rats the dose per injection was fixed at 2.0 mg/kg; for four others the dose per injection was varied irregularly, from one response to the next, between three levels (0.5, 1.0 and 2.0 mg/kg). Regardless of the dosing regimen, extracellular dopamine levels were tonically elevated by 200-800% within the cocaine self-administration periods, fluctuating phasically within this range between responses. In the fixed dose condition, the phasic increases following each injection (and the phasic decreases preceding them) averaged approximately 50% of the mean tonic elevation. Phasic fluctuations in dopamine levels remained time-locked to lever-presses even when response rate was irregular, because of the variable dose condition. In the variable dose condition greater increases in dopamine and longer inter-response times followed injections of the higher doses; dopamine fluctuations were consistent with the multiple-infusion pharmacokinetics of cocaine. DOPAC levels showed a slow tonic depression during cocaine self-administration, but individual injections were not associated with discernible phasic fluctuations of DOPAC. These data are consistent with the hypothesis that falling dopamine levels trigger successive responses in the intravenous cocaine self-administration paradigm, but inconsistent with the notion that extracellular dopamine levels are depleted at the times within sessions when the animal initiates drug-seeking responses. PMID- 7480532 TI - Defective habituation to nociceptive stimulation in alcohol-avoiding ANA rats. AB - Brain opioidergic mechanisms participate in the regulation of motivational and ingestive behaviours. Since alcohol is believed to activate endogenous opioid systems and to produce opioid-mediated antinociception, the present experiments were performed to find out if alcohol-induced antinociception differs between the alcohol-preferring AA and alcohol-avoiding ANA rat lines. Alcohol doses relevant to the voluntary alcohol intake by the AA rats (0.5-1.0 g/kg, intraperitoneally) failed to alter tail-flick (TF) latency in a 55 degrees C water bath by either rat line. However, repeated measurement of TF latency, even without any alcohol treatment, prolonged tail-flick latencies in AA but not in ANA rats. Prolongation of TF latency was also seen in non-selected Wistar rats, indicating that the ANA rats respond abnormally in this test. The antinociceptive effects of swimming induced stress (3 min at 15 degrees C) and those of cumulative morphine administration (0.5-16.0 mg/kg, subcutaneously) were similar in both rat lines. Using higher, motor-impairing alcohol doses with repeated baseline TF determinations, it was observed that a dose of 1.5 g/kg induced slight antinociception only in the AA rats, while 2.0 g/kg produced similar effects in both rat lines. It is thus concluded that the alcohol-preferring AA rats do not show any enhanced alcohol-induced antinociception at relevant alcohol doses. However, the alcohol-avoiding ANA rats appear to have a defective ability to habituate to repeated sensory stimuli, which could contribute to their alcohol avoidance by preventing the development of tolerance to aversive effects of alcohol. PMID- 7480533 TI - Localization of genes influencing ethanol-induced conditioned place preference and locomotor activity in BXD recombinant inbred mice. AB - Genetic differences in ethanol's ability to induce conditioned place preference were studied in 20 BXD Recombinant Inbred (RI) mouse strains and in the C57BL/6J and DBA/2J progenitor strains. Male mice from each strain were exposed to a Pavlovian conditioning procedure in which a distinctive floor stimulus (CS+) was paired four times with ethanol (2 g/kg). A different floor stimulus (CS-) was paired with saline. Control mice were injected only with saline. Floor preference testing without ethanol revealed significant genetic differences in conditioned place preference, with some strains spending nearly 80% time on the ethanol paired floor while others spent only 50% (i.e., no preference). Control mice showed genetic differences in unconditioned preference for the floor cues, but unconditioned preference was not genetically correlated with conditioned preference. There were also substantial genetic differences in ethanol-stimulated activity, but contrary to psychomotor stimulant theory, ethanol-induced activity on conditioning trials was not positively correlated with strength of conditioned place preference. However, there was a significant negative genetic correlation (r = -0.42) between test session activity and preference. Quantitative trait loci (QTL) analyses showed strong associations (P < 0.01) between conditioned place preference and marker loci on chromosomes 4, 8, 9, 18 and 19. Weaker associations (0.01 < P < 0.05) were identified on several other chromosomes. Analysis also yielded several significant QTL for unconditioned preference, ethanol-stimulated activity, and sensitization. Overall, these data support the conclusion that genotype influences ethanol-induced conditioned place preference, presumably via genetic differences in sensitivity to ethanol's rewarding effects. Moreover, several chromosomal regions containing candidate genes of potential relevance to ethanol-induced conditioned place preference have been identified. PMID- 7480534 TI - Conditioned reinforcing effects of capsules associated with high versus low monetary payoff. AB - The ability of a placebo drug capsule to serve as a conditioned reinforcer as a function of being paired with money reinforcement was evaluated. Volunteers were administered two differently colored capsules that presumably contained two different drugs. Although the volunteers were told they might contain a stimulant, sedative, or placebo, both capsules contained only a placebo. During sessions, volunteers participated in performance tasks. The tasks were programmed so that following one capsule, the amount of money obtained contingent upon responding was greater (high frequency of reinforcement) than following the other capsule (low frequency of reinforcement). During experiment 1, participants were exposed twice each to the two reinforcement conditions (sampling). During these choice sessions, 9 of 12 participants chose the capsule associated with the high frequency of reinforcement 2 or 3 times. Experiment 2 was designed to explore further whether the differential mood effects observed during sampling sessions could be conditioned. Although this could not be demonstrated, the self administration results demonstrating the control of choice behavior even in the absence of pharmacological effects suggest that drugs may function as conditioned reinforcers. This finding has implications for broadening our understanding of the determinants of initiation and continued drug use. PMID- 7480535 TI - Effects of post-ethanol administration of NMDA and non-NMDA receptor antagonists on the development of ethanol tolerance in C57B1 mice. AB - The effect of post-ethanol administration of NMDA and non-NMDA receptor antagonists on the development of environment-dependent ethanol tolerance was studied in C57B1 mice. Ethanol tolerance was produced by daily injections of ethanol (3.5 g/kg, IP) in the same experimental environment and measured as ethanol-produced "sleep-time" during 5 consecutive days. The non-competitive NMDA receptor antagonist, dizocilpine (MK-801; 0.1 mg/kg, IP), and the competitive NMDA receptor antagonist, CGP 39551 (5 mg/kg, IP), both given 120 min after the administration of ethanol, inhibited the development of tolerance to the hypnotic actions of ethanol. In contrast, the development of ethanol tolerance was not altered by administration of the specific AMPA/KA receptor blocking agents, NBQX (10 mg/kg, IP), and LY326325 (2.5 mg/kg, IP), respectively. Modulation of NMDA receptor activity by drugs like NMDA, d-cycloserine, and milacemide, which are known to enhance learning and memory in rodents, had no significant effect on the development of ethanol tolerance. Our present data confirm and extend previous findings which indicate that NMDA, but not non-NMDA, glutamate receptors may play an important role in the neuroadaptive processes associated with the development of ethanol tolerance. PMID- 7480536 TI - Differential modulation of antipredator defensive behavior in Swiss-Webster mice following acute or chronic administration of imipramine and fluoxetine. AB - The Mouse Defense Test Battery (MDTB) has been designed to assess defensive reactions in Swiss-Webster mice to situations associated with a natural predator, the rat. Primary measures taken before, during and after predator confrontation comprise escape attempts, predator assessment, defensive attack and flight. Previous reports from this laboratory have shown that the panic-promoting drug yohimbine potentiated flight behavior, while long-term treatment with the panicolytic agent alprazolam reduced this response. In order to evaluate further the possibility that the MDTB may represent an effective animal model of panic attacks, the present study investigated the behavioral effect of imipramine and fluoxetine, two serotonin reuptake inhibitors (SRIs) known to alleviate panic symptoms when given on a repeated basis. Both drugs were administered acutely and chronically (one daily IP injection for 21 days) at 5, 10 and 15 mg/kg. Our results showed that a single dose of imipramine or fluoxetine strongly potentiated flight reactions in response to an approaching predator and increased defensive attack toward the rat. This was in contrast to chronic treatment with each drug which dramatically decreased flight responses and defensive attack behaviors. In addition, long-term administration with both SRIs produced a reliable attenuation of predator assessment activities. Taken together, these findings suggest an acute anxiogenic-like effect of imipramine and fluoxetine followed by a fear/anxiety reducing effect after repeated administrations. These results support clinical observations revealing an acute anxiogenic effect of SRIs followed by an anxiolytic and/or panicolytic effect after chronic use, and support previous results suggesting that the MDTB may be useful for the investigation of panic-modulating agents. PMID- 7480538 TI - Contrasting effects on methamphetamine sensitization of ceruletide, a cholecystokinin-like decapeptide, and haloperidol. AB - Ceruletide, a cholecystokinin-like decapeptide, and haloperidol show neuroleptic actions through inhibition of dopamine release and blockade of dopamine receptors, respectively. In this study, the effects of both drugs on methamphetamine sensitization were assessed by means of ambulation in mice. The enhancement in ambulation increase caused by five repeated administrations of methamphetamine (2 mg/kg, SC) at 3- to 4-day intervals was dose-dependently reduced when it was administered simultaneously with ceruletide (0.01-0.1 mg/kg, SC) or haloperidol (0.03-0.3 mg/kg, SC). However, only haloperidol could inhibit the induction of methamphetamine sensitization as assessed by challenge with methamphetamine alone. Post-treatment with ceruletide (0.03 mg/kg) 3 h after each methamphetamine accelerated, whereas such post-treatment with ceruletide (0.1 mg/kg) or haloperidol (0.03-0.3 mg/kg) delayed, the induction of methamphetamine sensitization. On the other hand, mice given five pretreatments with ceruletide (0.01-0.1 mg/kg) or haloperidol (0.03-0.3 mg/kg) at 3- to 4-day intervals did not exhibit any significant change in the sensitivity to methamphetamine. The present results suggest that, in contrast to the dose-dependent inhibition of methamphetamine sensitization in the simultaneous administration and post treatment schedules, although both drugs can antagonize the acute stimulant effect of methamphetamine. PMID- 7480537 TI - Antipsychotic agents antagonize non-competitive N-methyl-D-aspartate antagonist induced behaviors. AB - Antipsychotic agents were tested for their ability to antagonize both dopaminergic-induced and non-competitive N-methyl-D-aspartate (NMDA) antagonist induced behaviors. All of the agents dose-dependently antagonized the apomorphine induced climbing mouse assay (CMA) and dizocilpine (MK-801)-induced locomotion and falling assay (MK-801-LF) with a CMA/MK-801-LF ratio of less than or equal to 1.6. However, clozapine and its structural analog olanzapine more potently antagonized MK-801-LF (1.1 and 0.05 mg/kg) than the CMA (12.3 and 0.45 mg/kg) and as a result had a CMA/MK-801-LF ratio of 11.2 and 9, respectively. Furthermore, phencyclidine (PCP) (2 mg/kg) can selectively induce social withdrawal in naive rats that were housed in pairs (familiar) for 10 days prior to testing without affecting motor activity. SCH 23390, raclopride, haloperidol, chlorpromazine and risperidone failed to reverse the social withdrawal induced by PCP up to doses which produced significant motor impairment. However, clozapine (2.5 and 5.0 mg/kg) and olanzapine (0.25 and 0.5 mg/kg) significantly reversed this social withdrawal in rats. Therefore, the non-competitive NMDA antagonists PCP and MK 801 can induce behaviors in Rodents which are selectively antagonized by clozapine and olanzapine. Furthermore, assessment of the effects of antipsychotic agents in the CMA, MK-801-LF and PCP-induced social withdrawal assays may provide a preclinical approach to identify novel agents for negative symptoms and treatment resistant schizophrenia. PMID- 7480539 TI - The effects of repeated treatment with 8-hydroxy-2-(di-n-propylamino) tetralin (8 OH-DPAT) on the lever press responding of the rat under FI and DRL schedules of food reinforcement. AB - In general, the effects of 8-OH-DPAT on the body temperature of rats or in inducing the 5-HT syndrome show rapid tolerance. However, in contrast, the 8-OH DPAT-induced increase in the activity of rats in a two-way active avoidance task only occurs after repeated administration, i.e. there is sensitisation. The present study was conducted to examine whether this developing hyperactivity may also be expressed as increased rates of lever press responding, and if so, under which conditions it occurs. Rats were trained to press levers under fixed interval 60-s (FI 60) or differential reinforcement of low rates 20-s or 72-s (DRL20, DRL72) schedules of food reinforcement. Groups of trained rats were then treated daily 5 min before testing with doses of 0.01, 0.1 and 1.0 mg/kg 8-OH DPAT SC for 10-21 days. In all three procedures, in the first couple of days of drug treatment, 8-OH-DPAT generally suppressed lever pressing in a dose-dependent manner. Thereafter, tolerance to this effect was seen to a greater (DRL20, DRL72) or lesser (FI60) extent. Some evidence for stimulation of low rates of lever press responding was seen after 10 days treatment under FI60, but not in DRL20 or DRL72 during short 30 to 60 min long daytime tests although in the latter case, the rats responded to the stimulating effects of 0.8 mg/kg SC amphetamine administered once at the end of the experiment. However, when rats were allowed to respond under DRL72 testing for 12 h during the night, after 10 days treatment a clear stimulation of lever pressing was observed. This stimulation was not specific to lever pressing, however, since a stimulation of entries into the food tray and licking were also seen. From these results, it may be concluded that the stimulating effect of 8-OH-DPAT after repeated administration may be expressed as increased rates of lever pressing, but not under all conditions in which psychomotor stimulation by amphetamine is seen. The potential for 8-OH-DPAT and related compounds to stimulate motor responding in this way should be taken into account when interpreting the effects of these drugs in animal models of psychiatric disorders. PMID- 7480540 TI - Effect of (-)-DS 121 and (+)-UH 232 on cocaine self-administration in rats. AB - The novel dopamine autoreceptor antagonists (-)-DS 121 and (+)-UH 232 were tested for their ability to alter cocaine self-administration behavior in rats reinforced on a progressive ratio (PR) schedule. (-)-DS 121 (15 mg/kg) and (+)-UH 232 (30 mg/kg) produced significant decreases in breaking point. (-)-DS 121 produced variable results on rate of cocaine intake on an FR1 schedule, indicating that rate may on occasion be insensitive to changes in cocaine reinforcement. In animals previously trained to self-administer cocaine, (-)-DS 121 failed to maintain responding when substituted for cocaine. This profile suggests that (-)-DS 121 is a promising new candidate for the treatment of cocaine abuse. PMID- 7480541 TI - Dissociation between the attentional effects of infusions of a benzodiazepine receptor agonist and an inverse agonist into the basal forebrain. AB - The effects of infusions of the benzodiazepine receptor (BZR) full agonist chlordiazepoxide (CDP) or the full inverse agonist beta-CCM into the basal forebrain on behavioral vigilance were tested. Vigilance was measured by using a previously characterized task that requires the animals to discriminate between visual signals of variable length and non-signal events. Measures of performance included hits, misses, correct rejections, false alarms, side bias, and errors of omission. Following the infusion of saline (0.5 microliters/hemisphere), the relative number of hits varied with signal length. In response to shorter signals, the number of hits decreased over time, indicating a vigilance decrement. Infusions of CDP (20, 40 micrograms/hemisphere) initially decreased the relative number of hits in response to shorter signals and, later in the course of the test sessions, to longer signals as well. CDP did not affect the relative number of correct rejections. In contrast, infusions of the inverse agonist beta-CCM (1.5, 3.0 micrograms/hemisphere) did not affect the relative number of hits but decreased the relative number of correct rejections (i.e., increased the number of false alarms). These data suggest that the basal forebrain mediates the attentional effects of BZR ligands. As systemic or intrabasalis administration of BZR agonists and inverse agonists was previously demonstrated to decrease and augment, respectively, activated cortical acetylcholine (ACh) efflux, their effects on behavioral vigilance are hypothesized to be mediated via their effects on cortical ACh. PMID- 7480543 TI - Catalepsy as a rodent model for detecting antipsychotic drugs with extrapyramidal side effect liability. AB - The predictive validity of catalepsy as a rodent model for detecting the extrapyramidal side effects (EPS) of antipsychotic drugs was recently questioned when the novel antipsychotic savoxepine produced little catalepsy in rodents while producing significant EPS in schizophrenic patients. Because catalepsy is viewed as an important model for predicting EPS, we decided to re-evaluate the effects of savoxepine. Savoxepine, clozapine, haloperidol, olanzapine, ORG 5222, raclopride, and risperidone were examined in two tests for catalepsy (grid and bar tests) in male Sprague-Dawley rats. The ability to antagonize amphetamine induced hypermotility was also examined, since this measure is believed to predict clinical efficacy. With the exception of clozapine, all drugs produced dose-dependent catalepsy in both tests. For each drug, the minimum effective dose for producing catalepsy was greater than or equal to the ED50 for antagonizing amphetamine-induced hyperactivity (defined as the dose producing a 50% reduction in hyperactivity). Clozapine resulted in the widest separation of effective doses in the catalepsy and activity models. Raclopride produced the next largest separation while the remaining drugs resulted in only a one- or two-fold dose separation between the two behavioral tests. The results with haloperidol and clozapine are consistent with the clinical effects of these drugs (severe versus mild EPS). The ratios of effective doses in catalepsy and activity for the remaining novel drugs are also consistent with preliminary clinical findings indicating some EPS with each of these compounds. Thus, catalepsy remains a suitable rodent model for detecting compounds with EPS liability in humans. PMID- 7480542 TI - Effects of single and repeated exposure to apomorphine on the acoustic startle reflex and its inhibition by a visual prepulse. AB - The acoustic startle reflex (ASR) is inhibited by startle-irrelevant stimuli that briefly precede reflex elicitation. This effect, prepulse inhibition (PPI), is reduced in strength for animals that have received dopamine agonists, such as apomorphine (APO). Reduction in PPI is most evident for weak masked noise prepulses, thus suggesting that APO disrupts the reception of stimuli to the extent that they present a low signal-to-noise ratio. Here we examine the effect of APO on PPI produced by non-masked visual prepulses. Light flashes were given at two intensities, 40, 70, 110, or 220 ms before ASR elicitation. In phase 1 (5 weeks in duration) half of the animals received one weekly injection of APO (0.5 mg/kg, IP) and one of vehicle (VEH), while the other half received two injections of VEH. Within these groups, half were tested 30 min after the injections, the other half kept test naive (four groups total). In phase 2, following a 4-week rest, all groups were tested after a low dose of APO (0.1 mg/kg) and VEH, 1 week apart. APO eliminated PPI for a dim flash and reduced PPI for a brighter flash to a level normally obtained with the dim flash, while increasing both ASR control values and activity. The bright light was maximally effective at a lead time of 70 ms and APO did not alter this value. Because in general the time of maximal inhibition varies with prepulse intensity for visual stimuli, the finding that the time of the peak remained constant reveals that APO has its effect on inhibition rather than on effective stimulus intensity. In phase 2, APO reduced PPI with no sign of sensitization from past drug exposure. However, APO increased the ASR only in groups previously exposed to APO, indicating behavioral sensitization. The differential effects of repeated exposure on these response measures suggest that neural substrates for the several behavioral effects of APO function at least in part independently. PMID- 7480544 TI - Further studies to examine the nature of dexfenfluramine-induced suppression of heroin self-administration. AB - The present series of experiments sought to investigate further the mechanism by which dexfenfluramine, a selective 5-HT releaser/reuptake inhibitor, reduces heroin self-administration by male Wistar rats. In experiment 1, the effect of combined intravenous heroin and intraperitoneal dexfenfluramine injections on operant responding for food was examined. In experiment 2, the maintenance of dexfenfluramine suppression of heroin self-administration following chronic (7 day) treatment was evaluated. Finally, in experiment 3, the ability of various 5 HT antagonists to block the dexfenfluramine suppression was examined. The results from experiment 1 suggest that sensorimotor deficits/malaise potentially associated with heroin/dexfenfluramine combinations are unlikely to account for the reductions in heroin self-administration. Experiment 2 suggested that the suppressant effect of dexfenfluramine on heroin responding may diminish rapidly following chronic treatment. Finally, central 5-HT1 and/or 5-HT2, but not 5-HT3, receptors may underlie the suppressant effects of dexfenfluramine on heroin self administration. PMID- 7480545 TI - Effect of scopolamine on visual attention in rats. AB - In the present study, the effects of scopolamine (SCOP) were determined upon the performance of rats in the five-choice serial reaction time task, a test of attention analogous to the continuous performance test in man. Rats were trained to detect and respond to brief flashes of light presented randomly in one of five locations until a stable level of performance was reached. SCOP (0.03-0.1 mg/kg SC) was administered 30 min prior to testing under standard conditions of stimulus presentation. SCOP reduced response accuracy at the highest dose and dose-dependently increased omissions and perseverative responses. However, these effects were mimicked by scopolamine methylbromide (SCOPMBr) which might suggest a peripheral site of action. When the task difficulty was increased by manipulating the stimulus presentation parameters, i.e. reduced stimulus intensity, duration or temporal predictability, SCOP (0.075 mg/kg SC) failed further to impair performance accuracy. However, in two separate experiments, SCOP (0.075 mg/kg SC) robustly increased the distractibility caused by a burst of loud white-noise occurring unpredictably during the intertrial-interval: SCOP significantly decreased accuracy and increased omissions, magazine latency, premature and perseverative responses compared with vehicle and the equivalent dose of SCOPMBr. These findings provide further evidence of a role for the central cholinergic system in attention. PMID- 7480547 TI - The stimulation of cholecystokinin receptors in the rostral nucleus accumbens significantly antagonizes the EEG and behavioural effects induced by phencyclidine in rats. AB - The influence of cholecystokinin (CCK), bilaterally injected into the rostral nucleus accumbens, on the EEG and behavioural effects induced by phencyclidine (PCP) has been studied in rats. CCK (10 ng) significantly inhibited PCP-induced EEG effects (increase of spectral power with respect to pre-drug tracing; increase of relative power distribution in the slowest frequency bands), and behavioural effects (circling and ataxia). The inhibitory effects of CCK were completely antagonized by 1 ng PD 135-158, a selective CCKB receptor antagonist, but not by lorglumide (1 microgram), a selective CCKA receptor antagonist. Since the effects induced by PCP in rodents have been proposed to be an experimental correlate of the psychotic symptoms it induces in humans, these results indicate that CCK may act as a neuroleptic. They also suggest that CCKB receptors located in the rostral nucleus accumbens may be involved in the neuroleptic-like activity of CCK. PMID- 7480546 TI - Cocaine and cocaethylene: effects on extracellular dopamine in the primate. AB - Cocaine and cocaethylene (a psychoactive metabolite of concurrent cocaine and ethanol consumption) were studied in the anesthetized vervet monkey. The ability of each to elevate extracellular DA in the caudate nucleus was assessed using microdialysis probes acutely lowered through chronic guide cannulae. Blood samples were also collected to determine plasma levels of the two drugs. Doses of 1.5 mumol/kg cocaine (equivalent to 0.5 mg/kg cocaine-HCl) and cocaethylene were administered intravenously. Microdialysis and blood samples were collected at 5 min intervals immediately following drug administration. Both drugs caused a maximal four-fold increase in extracellular DA during the 5- to 10-min period following drug administration. This is the first report of cocaine (and cocaethylene) induced alterations in extracellular DA in primates. The abilities of cocaine and cocaethylene to produce euphoria are being compared in ongoing clinical research studies. The potential use of these results for interpreting the neurochemical basis of any differences in those studies is discussed. PMID- 7480548 TI - Learning versus performance effects of cocaine on discriminative heart rate conditioning in rats. AB - The study examined the effects of cocaine on learning and performance of a classically conditioned heart rate (HR) discrimination in rats involving two auditory conditioned stimuli (CSs). In the discrimination protocol, one CS (CS+) was paired with the shock unconditioned stimulus (US) on a consistent basis and the other CS (CS-) was always presented alone. Four groups received an IP injection of 1, 3, 10, or 30 mg/kg cocaine and a fifth group received saline. Shortly after the injections, all groups were given six CS-alone trials, followed by 24 randomly sequenced discrimination conditioning trials (12 CS+ and 12 CS-). Approximately 72 h later, all groups were given six test trials with each CS in the absence of cocaine to evaluate the presence or absence of discrimination learning. All cocaine groups showed impaired discrimination performance on the discrimination conditioning trials, reductions in early pretest CS-alone responses, and reductions in resting HR. However, on the non-drug test trials discrimination performance was normal in all cocaine groups. The results established that in spite of major changes in HR dynamics, learning of the HR discrimination was not affected by cocaine but that cocaine did interfere with the performance of the discrimination. Except for the highest 30 mg group, the performance decrement appeared to be related to a cocaine-produced reduction in the capacity to inhibit bradycardia responding to the safe CS-. It was suggested that this loss of inhibitory control may have been due to cocaine changes in a corticothalamic pathway that controls inhibition of bradycardia to a safe CS-. PMID- 7480549 TI - Comparison of the spectrum of cognitive effects of alprazolam and adinazolam after single doses in healthy subjects. AB - Single doses of alprazolam (0, 0.5, 1.5 mg) or adinazolam mesylate sustained release tablets (SR) (0, 15, 45 mg) were administered to separate groups of 12 healthy men in a crossover design. Psychomotor performance was assessed by digit symbol substitution (DSST), and memory was assessed using a test battery which reflects various aspects of memory, including attention/working memory, explicit memory (recall of categorically related words), semantic memory (fragmented picture recognition, generation of category exemplars), and implicit memory (time saved in resolving fragmented pictures on the second exposure). Maximal psychomotor performance and memory decrements for the highest active doses were significantly different from placebo for all tasks at some time after dosing. The maximum decrement in DSST was not significantly different between drugs at the high dose (P = 0.288). Maximum attention/working memory decrements were significantly different between the high doses of the active compounds (P = 0.031), and the difference in maximum category recall decrement was marginally significant (P = 0.067). Access to knowledge memory was not significantly altered by these drugs; these results are similar to those obtained for other benzodiazepines. Both drugs exhibited slight effects on implicit memory. The results suggest that the sedative and memory effects of these triazolobenzodiazepines may not be closely related and suggest that adinazolam has a somewhat different spectrum of cognitive effects relative to alprazolam. PMID- 7480551 TI - Naloxone blocks the antianxiety but not the motor effects of benzodiazepines and pentobarbital: experimental studies and literature review. AB - The role of opioid systems in the anticonflict effect of chlordiazepoxide, diazepam and pentobarbital was evaluated with a modified Vogel procedure. First, morphine, ineffective by itself, was combined with subeffective or marginally effective doses of the benzodiazepines in order to detect possible potentiation. However, the combined treatment reduced licking in the Vogel procedure as well as in a licking test where no shock was administered. Several doses of the benzodiazepines and pentobarbital were then administered in combination with several doses of the opiate antagonist naloxone. A dose-dependent inhibition of anticonflict effect was obtained. In an additional experiment, it was shown that naloxone blocked the effects of diazepam in the elevated plus-maze procedure. Motor deficiencies, as evaluated with a rotarod test, produced by the benzodiazepines and pentobarbital could not be antagonized by naloxone. It is concluded that opioids are important for the anticonflict but not for the motor effects of these drugs. An analysis of published studies concerning the interaction of opioids and benzodiazepines in several procedures supposed to reflect anxiolytic effects shows that the inhibition obtained with naloxone is reliable and not procedure specific. The mechanisms by which opiate antagonists produce this inhibition of anticonflict activity are not known. It is tentatively suggested that opioid activation associated with stress may be a necessary component of anxiolysis. PMID- 7480550 TI - The delta opioid receptor antagonist naltrindole attenuates both alcohol and saccharin intake in rats selectively bred for alcohol preference. AB - This study demonstrates that the selective delta receptor antagonists ICI 174864 and naltrindole (NTI) attenuate alcohol intake in a dose-dependent manner, without altering water intake, in rats selectively bred for alcohol preference. ICI 174864 had a very limited duration of action, as evidenced by the fact that suppression of alcohol intake lasted for only an hour following ICI 174864 administration. NTI, when administered in a dose of 10 mg/kg, suppressed alcohol intake by 28%. Increasing the dose of NTI to 15 mg/kg produced a 44% suppression of alcohol intake, but a further increase to 20 mg/kg did not produce greater suppression than was seen with a dose of 15 mg/kg (46% versus 44%, respectively). This suggests that NTI is maximally effective in suppressing alcohol intake at a dose of 15.0 mg/kg. NTI displayed a long duration of action, as evidenced by attenuation of alcohol drinking that lasted for at least 8 h following drug treatment. Administering the maximally effective dose of NTI (15 mg/kg) in two parts, separated by 4 h, served to prolong the duration of action of NTI and produced an attenuation of alcohol intake, but not water intake, that lasted for at least 28 h. The effect of NTI on alcohol intake was not specific for alcohol, as evidenced by the fact that NTI reduced the intake of saccharin solutions with and without alcohol. PMID- 7480552 TI - Reward shifts and motor responses following microinjections of opiate-specific agonists into either the core or shell of the nucleus accumbens. AB - Differences in pharmacology, anatomical connections, and receptor densities between the "core" and "shell" of the nucleus accumbens suggest that behavioral activity normally modulated by the accumbens, such as reward and motor functions, may be differentially regulated across the mediolateral axis. This study investigated the effects of opiate receptor-specific agonists on reward and motor functions in either the accumbens core or shell, using the intracranial self stimulation (ICSS) rate-frequency curve-shift method. Microinjections of the mu opiate receptor-specific agonist, DAMGO (vehicle, 0.03 nmol, and 0.3 nmol), or the delta opiate receptor-specific agonist DPDPE (vehicle, 0.3 nmol, 3.0 nmol), were administered bilaterally in a random dose order with a minimum of 3 days between injections. Rats were tested over three consecutive 20-min rate-frequency curves immediately following a microinjection to investigate the time course of drug effects. Both opiate agonists decreased the ICSS frequency necessary to maintain half-maximal response rates when injected into the medial and ventral shell region of the accumbens. However, DAMGO microinjections into the lateral accumbens core or the control site of the caudate increased the frequency necessary to elicit half-maximal response rates, while DPDPE microinjections into these regions had no effect. Evaluation of motor effects show that administration of DAMGO resulted in a suppression of activity in all locations. In contrast, DPDPE microinjections resulted in little or no effect on lever pressing activity at any location. PMID- 7480553 TI - Phencyclidine injections into the dorsal hippocampus disrupt long- but not short term memory within a spatial learning task. AB - Since the hippocampus is likely to be a major site of phencyclidine (PCP) action, the effects of various doses of PCP (1.8, 18 or 36 nM) as well as 3.6 nM MK-801 or saline injected directly into the dentate gyrus of the hippocampus was tested for acquisition of a spatial navigation task (dry land version of a water maze) using a paradigm that assesses short term memory based on learning within a day and long term memory based on learning between days. Results indicated that relative to saline or 1.8 nM PCP injected rats, rats with 18 or 36 nM PCP or 3.6 nM MK-801 injections were impaired in acquisition of the task as measured by increased distances traveled to find the food location between days but not within days. In additional experiments 36 nM PCP or 3.6 nM MK-801 did not produce any deficits in the acquisition of an object discrimination task. It is suggested that PCP through its blocking action of the NMDA receptor in the dentate gyrus or CA1 region of the dorsal hippocampus mediates the consolidation of new spatial location information. PMID- 7480554 TI - Smoking, processing speed and attention in a choice reaction time task. AB - Nineteen subjects performed a choice reaction time task in which two levels of choice (two and four stimuli), and two levels of spatial attention (narrow and wide) were manipulated under each of two smoking conditions: sham smoking (denicotinised cigarette) or regular smoking (0.8 mg nicotine cigarette). All three factors significantly affected reaction time, with the smallest reaction times being recorded to the two-choice narrow grouped stimuli recorded under the high nicotine condition. Nicotine appears to speed decision time for both complex and hard-to-attend tasks, which is compatible with a role for nicotinic receptors in systems jointly mediating attention, memory and processing speed. PMID- 7480555 TI - The role of the ascending 5-hydroxytryptaminergic pathways in timing behaviour: further observations with the interval bisection task. AB - This experiment examined the effect of destroying the 5-hydroxytryptaminergic (5HTergic) pathways on rats' ability to discriminate between two durations. Rats received injections of 5,7-dihydroxytryptamine into the median and dorsal raphe nuclei or sham lesions. They were trained to press lever A following a 2-s presentation of a light and lever B following an 8-s presentation of the light. For some rats, the levers were inserted into the chamber immediately after stimulus presentation ("no-poke-requirement"); for others, the levers were not inserted until a flap covering the food tray positioned midway between the levers had been depressed ("poke-requirement"). When stable performance was attained, "probe" trials were introduced in which the light was presented for intermediate durations. Logistic functions were derived relating percent choice of lever B to log stimulus duration. Under the "no-poke-requirement" condition, the bisection point (duration yielding 50% choice of lever B) was shorter in the lesioned rats than in the control rats. Under the "poke-requirement" condition, this effect of the lesion was attenuated. There was no effect of the lesion on the Weber fraction under either condition. The levels of 5HT and 5-hydroxyindoleacetic acid were reduced in the brains of the lesioned rats, but the levels of noradrenaline and dopamine were not altered. It is proposed that rats may attain accurate timing under the interval bisection task by moving from one lever to the other during stimulus presentation; this movement may be facilitated by destruction of the 5HTergic pathways.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7480556 TI - Differential effects of flumazenil in alcoholic and nonalcoholic cirrhotic patients. AB - Ligands to the benzodiazepine receptor (BZR) accumulate in hepatic encephalopathy; the benzodiazepine antagonist flumazenil can reverse some manifestations of this condition. This study was designed to explore the effects of flumazenil on cognitive function and anxiety levels in cirrhotic patients without hepatic encephalopathy. Twenty such patients--ten alcoholic, ten nonalcoholics--and ten normal volunteers matched for age and sex were randomly allocated to treatment order (flumazenil or placebo first) in a double-blind cross-over trial. Cognitive function was evaluated with a battery of psychological tests shown previously to be sensitive to the impairment induced by liver disease. Cirrhotic patients performed worse than controls on several tests: digit cancellation, digit symbol substitution, key tapping and Reitan's trail B test. Flumazenil did not reverse these cognitive impairments but it did induce anxiety in nonalcoholic cirrhotics. On one index of memory--delayed word recall- alcoholics performed worse than nonalcoholic cirrhotics. Flumazenil reversed this memory impairment in the alcoholic cirrhotic group. These results suggest that alcohol consumption induces changes at the BZR that are different from changes induced solely by liver impairment. PMID- 7480558 TI - Society for Psychophysiological Research 35th annual meeting, Toronto, Canada, October 12-15, 1995. Abstracts. PMID- 7480557 TI - Effects of the dopamine D1 receptor antagonist SCH 39166 on the ingestive behaviour of alcohol-preferring rats. AB - The present study evaluated the effect of the selective and long-acting dopamine D1 receptor antagonist SCH 39166 on several aspects of the ingestive behaviour of genetically selected alcohol-preferring rats, bred from Sardinian alcohol preferring rats. The effect of subchronic (8 days) subcutaneous drug administration was evaluated on the simultaneous daily intake of 10% ethanol, food and water. SCH 39166, 0.1 mg/kg, did not significantly modify the intake of the three ingesta. The dose of 1 mg/kg differentially modified rat ingestive behaviour, inhibiting intake of 10% ethanol, without modifying total fluid and food intake. The higher dose of 5 mg/kg produced a non-selective suppression of ingestive behaviour, which was accompanied by behavioural impairment. Acute drug injection was tested on 2-h intake of 10% sucrose, 0.1% saccharin, water or food. The doses of 0.1 and 1 mg/kg markedly inhibited the 2-h intake of 10% sucrose and 0.1% saccharin, but they did not modify either the 2-h water intake in water deprived and water sated rats or the 2-h food intake in food deprived rats. These findings suggest an important role of mechanisms mediated by D1 receptors in the control of alcohol and sweet solution intake, but not in water and food intake. Moreover, they indicate that SCH 39166, in relation to its selectivity and long lasting activity, is an interesting pharmacological tool to investigate further the role of D1 receptor mechanisms in the control of ingestive behaviour. PMID- 7480559 TI - A tale of two societies. AB - Although consultation-liaison psychiatry grew out of the general hospital psychiatry movement, the American Psychosomatic Society has played a vital role in nurturing the discipline. Through the Society's journal, Psychosomatic Medicine, and its meetings, consultation psychiatry continues to stay in the mainstream. The subspecialty has developed in both content and strength to merit a focused organization, the Academy of Psychosomatic Medicine. The role of both organizations in the development of consultation-liaison psychiatry is discussed and recommendations for future collaborative activities are suggested. PMID- 7480560 TI - Physical symptoms of chronic fatigue syndrome are exacerbated by the stress of Hurricane Andrew. AB - This study examined the effects of Hurricane Andrew on physical symptoms and functional impairments in a sample of chronic fatigue syndrome (CFS) patients residing in South Florida. In the months after Hurricane Andrew (September 15 December 31, 1992), 49 CFS patients were assessed for psychosocial and physical functioning with questionnaires, interviews, and physical examinations. This sample was made up of 25 CFS patients living in Dade county, a high impact area, and 24 patients in Broward and Palm Beach counties, areas less affected by the hurricane. Based on our model for stress-related effects on CFS, we tested the hypothesis that the patients who had the greatest exposure to this natural disaster would show the greatest exacerbation in CFS symptoms and related impairments in activities of daily living (illness burden). In support of this hypothesis, we found that the Dade county patients showed significant increases in physician-rated clinical relapses and exacerbations in frequency of several categories of self-reported CFS physical symptoms as compared to the Broward/Palm Beach county patients. Illness burden, as measured on the Sickness Impact Profile, also showed a significant increase in the Dade county patients. Although extent of disruption due to the storm was a significant factor in predicting relapse, the patient's posthurricane distress response was the single strongest predictor of the likelihood and severity of relapse and functional impairment. Additionally, optimism and social support were significantly associated with lower illness burden after the hurricane, above and beyond storm-related disruption and distress responses. These findings provide information on the impact of environmental stressors and psychosocial factors in the exacerbation of CFS symptoms. PMID- 7480561 TI - Macronutrient intake, eating habits, and exercise as moderators of menstrual distress in healthy women. AB - The present investigation studied the relationship between symptoms of menstrual distress and macronutrient intake, eating behavior, and exercise in healthy women. Twenty-six normally menstruating women with no complaints of menstrual distress completed a disguised questionnaire on menstrual symptoms and monitored the type and amount of food consumed as well as the type and duration of exercise during a full menstrual cycle. Menstrual cycle phases were determined by the presence of menses, ovarian hormonal assays, and basal temperature monitoring. Reports of pain, water retention, negative affect, behavior change, and arousal were significantly higher (p < .05 or better) in the perimenstruum when compared to the follicular and luteal phases. During the perimenstruum, a higher energy intake of carbohydrate was associated with higher ratings of negative affect (p < .01) and impaired performance/decreased activity (p < .05). Lower energy intake of protein was associated with higher ratings of well being (p < .05). Overeating and dieting behavior were related to greater water retention (p < .01), autonomic reactions (p < .05), and appetite (p < .05). The amount of aerobic exercise in contrast to the intensity was related to lower water retention (p < .01), autonomic reactions (p < .05), and appetite (p < .01). Carbohydrate consumption, eating behavior, and regular exercise are reliably associated with menstrual distress and deserving of experimental evaluation as treatment interventions for menstrual distress. PMID- 7480562 TI - Power spectral analysis of heart rate variability in healthy young women during the normal menstrual cycle. AB - This study investigated the fluctuations of autonomic nervous activities during the menstrual cycle. Twenty college females were tested for cardiovascular reactivity to mental challenge during both follicular and luteal phases across two menstrual cycles. Power spectral analysis of heart rate variability (HRV) was used to examine the autonomic nervous activities. At baseline, although heart rate and blood pressure did not differ across the menstrual cycle, the low frequency (LF) component in the HRV was higher and the high-frequency (HF) component in the HRV was lower during the luteal phase than during the follicular phase. The LF/HF ratio was also significantly greater in the luteal phase. These data suggest that sympathetic nervous activities are predominant in the luteal phase as compared with follicular phase. In addition, the power spectral analysis of HRV has more sensitivity than heart rate or blood pressure in assessing the slight fluctuations of autonomic nervous activities during the menstrual cycle. PMID- 7480563 TI - Stress and the erectile response to intracavernosal prostaglandin E1 in men with erectile dysfunction. AB - The erectile response to intracavernosal injection (ICI) of smooth muscle relaxants is often impaired in men with presumed psychogenic erectile dysfunction. This study tests the hypothesis that such impairment results from stress-related increase in circulating norepinephrine (NE). Fifty-nine men with erectile dysfunction had their nocturnal penile tumescence (NPT) monitored over 2 nights, and ICI of 10 micrograms prostaglandin E1 was given on the second morning. Psychometric and neuroendocrine measures of stress, including plasma and urinary catecholamines, were taken on both mornings. Inhibition of the ICI response was determined by the difference between NPT and ICI responses, and two groups of "high inhibition" and "low inhibition" men were compared. The high inhibition group showed higher "Trait" and "State" anxiety and a neuroendocrine profile of higher initial cortisol and lower plasma catecholamine levels than the low inhibition group. These differences were evident on both mornings and hence were not specifically related to the ICI. There was no support for the hypothesis that inhibition of response to ICI results from increased circulating NE, though the possibility that increased NE release occurred specifically in the erectile tissues could not be excluded. PMID- 7480564 TI - The effects of a new alpha-2 adrenoceptor antagonist on sleep and nocturnal penile tumescence in normal male volunteers and men with erectile dysfunction. AB - The effects of a new alpha-2 adrenoceptor antagonist, RS 15385, on sleep and nocturnal penile tumescence (NPT) was assessed by intravenous infusion of the drug, in two doses (high and low), and of placebo through the night in 12 young normal volunteers and 24 men with erectile dysfunction. The drug reduced total sleep time in the younger normal volunteers only with the high dose and in the dysfunctional men with both doses. There was no selective effect on different sleep stages in the normal volunteers, except for an increase in the per cent of total sleep time in Stage 1 with the high dose. The dysfunctional men were divided into two age groups (< 47 years and > 47 years). There was an age-related increase in the proportion of total sleep in rapid eye movement (REM), which was only apparent in the older age group. In the volunteers, there was a curvilinear dose-response effect on NPT. The lower dose modestly increased erectile response, particularly during non-REM sleep, whereas the higher dose reduced erectile response, most noticeably during REM. The only positive effect of the higher dose was an increase in spontaneous erections after lights out before sleep onset. The older dysfunctional group showed no drug effects on NPT. The younger dysfunctional men showed increased erectile response during non-REM with the higher dose, an effect that was also significant in the interval between sleep onset (Stage 2) and first REM. PMID- 7480568 TI - Parental history of hypertension, sodium loading, and cardiovascular response to stress. AB - Parental history of hypertension, dietary sodium, and psychological stress have all been implicated in the development of essential hypertension and may interact in elevating disease risk. The mechanism by which this might occur is unclear, but it may be related to changes in the peripheral vasculature. The present study examined the effects of parental history and sodium on cardiovascular responses to an extended stressor. Eighteen normotensive offspring of hypertensives and 18 offspring of normotensives were exposed to a 1-hour shock-avoidance video-game procedure after 14 days of sodium loading (10 1-g tablets/day) and again after 14 days of placebo tablets. Order of sessions was counterbalanced between subjects in a double-blind design. In offspring of hypertensives, sodium loading elevated total peripheral resistance and norepinephrine responses to stress relative to placebo conditions and compared with offspring of normotensives. These increases were accompanied by decreases in stroke volume and cardiac output, which may explain the absence of familial differences in blood pressure responses to stress and sodium. Sodium loading had no effect on offspring of normotensives. The elevated resistance in offspring of hypertensives may suggest the initiation of pathological processes. The absence of sodium effects on resting values indicates the importance of research under conditions of stress. PMID- 7480565 TI - Gender differences in patterns of dynamic cardiovascular regulation. AB - The purpose of this research was to examine the role of gender in hemodynamic response patterns to stress. Sixty-four male and 55 female young adults were administered a protocol of rest, mental arithmetic, video game, and anger recall interview while blood pressure, heart rate, and cardiac impedance measures were obtained. Men had higher levels of systolic blood pressure and cardiac output and greater reactivity to tasks, indexed by increases in cardiac output and diastolic blood pressure. Extreme groups of myocardial and vascular reactors were formed from cardiac output and total peripheral resistance change scores during mental arithmetic. The myocardial reactors exhibited greater sensitivity to task demands, whereas vascular reactors exhibited comparable total peripheral resistance increases to all tasks. Vascular reactors exhibited greater diastolic blood pressure reactivity than myocardial reactors. Future studies should address the predictive validity of these reactor patterns for the subsequent development of coronary heart disease. PMID- 7480567 TI - Inhibited breathing decreases renal sodium excretion. AB - This study tested the hypothesis that breathing at the upper end of the normal range of end tidal CO2 decreases renal sodium excretion. Normotensive human subjects learned to self-regulate end tidal CO2 using a respiratory gas monitor and feedback procedure. Urine flow rates were increased by a standardized water drinking regimen. Urinary volume and sodium and potassium excretion were decreased during 30 minutes of inhibited (i.e. high normal end tidal CO2) breathing, compared with levels preceding and after task performance. Blood pressure, but not heart rate, increased during task performance. Plasma volume increase under these conditions is indicated by the observation that urinary excretion of an endogenous digoxin-like factor was increased. The physiological mechanism by which inhibited breathing elicits renal sodium retention remains to be determined. This breathing pattern could mediate the role of behavioral stress in some forms of hypertension. PMID- 7480569 TI - Self-efficacy as a moderator of perceived control effects on cardiovascular reactivity: is enhanced control always beneficial? AB - We have found that enhanced control has an attenuating effect on cardiovascular reactivity when effort of responding is maintained constant; however, not all individuals will react to increased control in the same manner. In the present study, 40 subjects engaged in a mental arithmetic task under high control (self paced) and low control (externally paced) conditions. Subjects' self-efficacy concerning this task was assessed. As expected, significant main effects were found for control condition, with high control producing smaller blood pressure and heart rate changes than low control (11.4 vs. 20.4 mm Hg (systolic blood pressure), 4.4 vs. 11.4 mm Hg (diastolic blood pressure), and 6.2 vs. 7.9 beats per minute (heart rate)). No main effects were found for self-efficacy. However, the interaction between control and self-efficacy was significant for systolic blood pressure and heart rate and marginally significant for diastolic blood pressure; post hoc tests showed that this was due to the effect of self-efficacy classification under high control conditions; subjects with low self-efficacy for the mental arithmetic task evidenced cardiovascular changes that were significantly greater than those of the high self-efficacy group (8.0 vs. 14.8 mm Hg (systolic blood pressure), 2.7 vs. 6.1 mm Hg (diastolic blood pressure), and 5.2 vs. 7.1 beats per minute (heart rate). The data suggest that the reactivity observed during active coping is due in part to the effort of responding and in part to the match between the demands of the task and certain mastery-related attributes of the individual. PMID- 7480566 TI - Adrenergic blockade ameliorates cellular immune responses to mental stress in humans. AB - This study evaluated the sympathoadrenal modulation of behaviorally evoked immune responses by administration of a nonselective adrenoceptor antagonist (labetalol) to subjects exposed to mental stress. In a 2 x 2 factorial design, subjects were assigned to a labetalol or saline condition and, within each condition, were exposed either to acute laboratory stress or no stress (control). Lymphocyte subsets, natural killer (NK) cell cytotoxicity, and T cell proliferation to phytohemagglutinin and concanavalin A were assessed pre-experimentally, at baseline after infusion and after 18 minutes of mental stress (or rest). By comparison with the other three conditions, the saline-stress group showed a greater peripheral NK cell number and cytotoxicity, lower mitogenic response to phytohemagglutinin and concanavalin A, and diminished ratio of CD4:CD8 cells after the stressor. As predicted, immune responses did not differ among the remaining groups (labetalol-stress, saline-rest, labetalol-rest). Group differences in NK cell cytotoxicity were not significant after controlling for differences in NK cell numbers. These findings demonstrate that the occurrence of certain immunologic responses to acute psychological stress are dependent on concomitant activation of the sympathetic nervous system. PMID- 7480570 TI - Relationships between thyroid hormones and symptoms in combat-related posttraumatic stress disorder. AB - This study was designed to investigate relationships between serum thyroid hormone levels and the severity of posttraumatic stress disorder (PTSD) symptoms in a group of 65 male Vietnam combat veterans who participated as members of cohorts in an elective inpatient treatment program. Thyroid hormone measures included serum free and total triiodothyronine (T3), free and total thyroxine (T4), and thyroxine-binding globulin. To estimate symptom severity, the Clinician Administered PTSD Scale (CAPS-2), based on DSM-III-R diagnostic criteria for PTSD, was used. Significant positive correlations were observed between free T3, total T3, total T4, and the "hyperarousal" frequency subscale score and the CAPS 2 frequency sum score. Patients with increased thyroid hormone levels and increased hyperarousal symptoms might constitute a clinically significant subtype among patients with PTSD. Alternatively, increased thyroid activity and hyperarousal symptoms may be associated with phase-related characteristics in PTSD. Research strategies for further evaluation of these preliminary findings are discussed. PMID- 7480571 TI - Metaphor and meaning in conversion disorder: a brief active therapy. AB - The concept of conversion disorder as a change in somatic function that symbolically represents an unconscious conflict is currently challenged in the literature. In this article, the author elaborates on the psychodynamic concept of conversion and defines its characteristics and mode of diagnosis. The usefulness of this approach is demonstrated in a detailed case presentation of a brief, active psychodynamic psychotherapy of six sessions that led to the rapid disappearance of symptoms. In particular, the metaphoric meanings of the physical symptoms were interpreted and followed by an immediate disappearance of symptoms. The logic of this therapeutic approach and its implications as a transference cure are discussed in detail. PMID- 7480572 TI - The stability of alexithymia in depressed patients. AB - The stability of alexithymia as measured by the Toronto Alexithymia Scale (TAS) and its relationship to depression were investigated in 50 depressed inpatients. The test-retest coefficient for TAS over a 5-day period was 0.57 (p < 0.001). A reliable change index for depressed mood indicated that mood covaried with the TAS, but changes in TAS were not clinically significant. The data support alexithymia as a stable personality construct. PMID- 7480573 TI - Influence of hereditary factors in psychogenic disorders. AB - As part of a research project, a sample of 50 pairs of twins (21 pairs of identical twins, 16 pairs of nonidentical twins of the same sex, and 13 pairs of male-female twins, n = 100 test persons) was examined between 1963 and 1969 and recently, 20 years later, followed up. The index twins were drawn from among the patients making use of the services of an outpatient clinic; they had been diagnosed as having psychoneurotic, character-neurotic of psychosomatic disorders. The question again looked into was that of nature versus nurture: identical twins displayed significantly higher similarity with regard to both severity of their neuroses and manifestation of neurotic symptoms than did nonidentical twins. The hereditary factor proved to be of greater import for the continuing childhood neuroses than for the subsiding childhood neuroses. Yet a certain hereditary factor effect was observed for the neuroses which first manifest themselves in adulthood. Our findings indicate that the hereditary factor is not more pronounced in childhood than in adulthood; it is equally effective in adults. PMID- 7480574 TI - A comparative study of thought disorder in acute and chronic schizophrenia. AB - A differential phenomenological study of acute and chronic schizophrenia is scanty. Thought disorder was assessed in 22 acute and 23 chronic schizophrenics. The scale for the assessment of thought, language and communication was used. Poverty of speech was significantly more frequent in acute schizophrenia. Positive formal thought disorder was unusually found to be severer in chronic schizophrenia. No other significant difference was found. From the perspective of thought disorder, acute and chronic forms of schizophrenia seem to be in a continuum with minimal difference. PMID- 7480575 TI - Cognitive performance in patients recovering from depression. AB - Investigations into the development of cognitive impairment in 30 hospitalized depressive patients have suggested that such disturbances encompass two dichotomous entities: a core entity of long-persisting, therapy-resistant impairment and an entity of reversible impairment with a prompt onset of improvement. The course of improvement turned out to evolve largely independently with respect to psychopathology: about one half of patients displayed severe cognitive dysfunctions which remained virtually unchanged until hospital discharge, while 80% significantly improved within the first 12 days with respect to their depressive symptoms, and 53.3% displayed a clear response to treatment at the time of hospital discharge. In particular, a considerable number of patients with a significant reduction of depressive symptomatology at hospital discharge still suffered from severe cognitive dysfunctions. Neither antidepressant or antipsychotic medication nor acute side effects due to medication explained the development of cognitive impairment. Accordingly, cognitive impairment seems to represent an essentially independent syndrome complex comparable to 'deficits' or 'negative symptoms' in schizophrenia. On the other hand, single case correlation analysis revealed a somewhat closer relationship of cognitive impairment with speech behavior and voice sound characteristics. Two thirds of patients displayed significant correlations between cognitive performance scores and second-order constructs like speech flow, dynamics or intonation. However, the respective subgroups defined by significant correlations with single speech parameters were relatively small (< or = 35% of cases), thus indicating that there exist large interindividual differences as to how cognitive impairment affects the patients' speech. PMID- 7480576 TI - Compulsive and impulsive symptomatology in trichotillomania. AB - Although classified as a disorder of impulse control, trichotillomania (hair pulling) may have some phenomenological overlap with obsessive-compulsive disorder (OCD). The question arises as to whether trichotillomania is best conceptualized as a disorder characterized by impulsivity or compulsivity. Impulsive and compulsive symptoms and traits were compared in 43 patients who presented for treatment of trichotillomania, OCD or impulsive personality disorder. Trichotillomania patients had significantly lower scores of obsessive compulsive symptoms than OCD patients, and significantly higher impulsiveness scores than this group. Measures of aggression did not differ significantly between groups. PMID- 7480577 TI - True auditory hallucinations as a conversion symptom. AB - Detailed descriptions of true hallucinations as a conversion symptom are rare in the psychiatric literature. The author presents a case with true conversive auditory hallucinations which paradigmatically demonstrates the usefulness of the concept in the treatment of such hallucinations. The phenomenon of true conversive hallucinations may contribute to the embryological understanding of mental disorders. PMID- 7480578 TI - Expressed emotion, communication deviance and schizophrenia. An exploratory study of the relationship between two family variables and the course and outcome of a psychoeducational treatment programme. AB - The relationship between expressed emotion (EE) and communication deviance (CD), and outcome in a group of early-onset schizophrenics after 2 years of psychoeducational treatment was examined. Patients' parents were assessed on EE and CD before and after the 2-year period. Outcome was measured by Global Assessment Scale (GAS) and relapse rate. Results showed that the correlation between CD and outcome was higher than between EE and outcome. In most of the patients who improved their GAS score during the treatment programme, their parents changed from high to low EE. CD proved to be more resistant to psychosocial intervention than EE. PMID- 7480579 TI - Beyond the brain-mind dichotomy and toward a common organizing principle of pharmacological and psychological treatments. PMID- 7480581 TI - Alexithymia: a further exploration of its nomological network. AB - The present studies were designed in order to obtain a better understanding of the nomological network of the alexithymia concept. In study I, the links between alexithymia, coping, and self-rated health were explored. As predicted, strong negative correlations were found between alexithymia and the expression of emotions, daydreams and fantasies, and planful and rational actions. Contrary to expectations, no clear associations were found with self-reported health status. In study II, the focus was on links with personality, temperament, and self reported (susceptibility to) homesickness. Although some correlations reached statistical significance, all were rather modest. The most remarkable finding was the independence of alexithymia and neuroticism. In study III, the associations between alexithymia and aspects of love experiences were examined, again yielding low correlations. It is concluded that alexithymia is relatively independent of temperament and personality. In addition, it is suggested that the association between alexithymia and the experience of positive emotions deserves further exploration. PMID- 7480580 TI - Work environment, skills utilization and health of Swedish nurses--results from a national questionnaire study. AB - Nurses make up a considerable proportion of the Swedish health care staff. Few nationally representative studies of their work environment and health, however, have been published. The present questionnaire study on work conditions, skill utilization, health and well-being of a national representative sample of 3,500 Swedish nurses, indicates a high degree of job satisfaction. Despite high satisfaction, 40% of the respondents felt too tired after work to socialize with family and 45% had seriously considered quitting their jobs. Factors such as improved work climate and development of supervisory skills were suggested as psychosocial interventions. Improvement in physical work environment factors had low priority. Medical care nurses perceive a higher work load and higher job demands in relation to decision latitude, and geriatric nurses report less work development. No differences between disciplines were found in work relations. In conclusion, there is a need to further develop skills development and utilization and work organization. Results will be used in interventions to enhance the work environment for nurses. PMID- 7480582 TI - Predictive value of alexithymia: a prospective study in somatizing patients. AB - In the present study, the potential role of alexithymia in predicting the long term treatment outcome was investigated prospectively in 30 patients with DSM-III R somatoform disorders and anxiety disorders. Using SCID interviews, diagnoses were assessed before inpatient treatment and 2 years after discharge. Patients who met criteria for DSM-III-R undifferentiated somatoform disorder at follow-up exhibited higher pretreatment alexithymia scores (as measured by the TAS) as compared with patients who showed remission of their somatoform disorder or patients who never had met criteria for a somatoform disorder. As a result stepwise logistic regression analyses, high alexithymia scores emerged as a significant predictor of persistent somatization, independent of other measures of psychopathology, sociodemographic variables, and measures of illness severity. PMID- 7480583 TI - Alexithymia in inflammatory bowel disease. A case-control study. AB - The purported association between alexithymia and inflammatory bowel disease (IBD) was investigated in a group of 112 IBD patients (89 with ulcerative colitis and 23 with Crohn's disease) using the well-validated 20-Item Toronto Alexithymia Scale. Alexithymia was assessed also in a group of 112 normal subjects matched for gender, age, and education. The IBD group was significantly more alexithymic than the control group, and no significant difference was found between the ulcerative colitis and Crohn's disease patients. Alexithymia was unrelated to the duration of illness and the level of disease activity. Although the rate of alexithymia in the IBD group (35.7%) was significantly higher than the rate in the control group (4.5%), it is lower than rates of alexithymia that have been found among psychiatric patients with disorders that also have been linked theoretically and clinically with alexithymia. PMID- 7480584 TI - Predictors of job satisfaction and job influence--results from a national sample of Swedish nurses. AB - Results from a questionnaire study on work conditions, job satisfaction, job influence, health and well-being, based on a representative sample of 3,500 Swedish nurses, indicate a high rate of overall job satisfaction. High decision latitude, consistency between work assignments and education, as well as low sickness absenteeism did all associate with high job satisfaction, number of working years did not. Compared to nurses satisfied with their jobs, twice as many sick days were found among those dissatisfied. Greater job influence related to lower job demands, more stimulating work and skills development as well as more responsibility and involvement. Anxiety and depression, intellectual satisfaction, job demands, job control and perceived general health explained 22% of the total variance in job satisfaction. The study has identified some key factors influencing overall job satisfaction. By focusing on these, job satisfaction among nurses should improve. PMID- 7480585 TI - [Assessment of grief exemplified pregnancy loss: development and initial results on the validity of the Munich Grief Scale]. AB - Bereavement research is hampered conceptually by confounding of grief and depression, methodologically by a lack of generally agreed-upon measures. Therefore we developed the Munich Grief Scale (MGS) based on the Perinatal Grief Scale (PGS, Potvin et al. 1989) and results from a previous study. Validation included comparisons with standardized self-report scales of depression, anxiety and physical symptoms. It was based on a follow-up study of 125 patients who had miscarried and 3 comparison groups who either suffered from a miscarriage or a stillbirth. Self-reports were compared to expert ratings of grief and depression. The MGS is a economical, reliable self- and expert rating scale after pregnancy loss. The applicability of the grief measures to other kinds of bereavements remains to be determined. PMID- 7480586 TI - [Study of the practicability of qualitative and quantitative documentation of outcome in inpatient psychotherapy]. AB - The results of a study evaluating the practicability of two newly-developed documentation forms are presented (n = 336 inpatient psychosomatic psychotherapeutic treatment episodes in 8 clinics). Parallel versions of the forms were completed by patients and therapists. The "ErgeDoku-A-Form" allows for the naming of up to five therapy goals determined at the beginning of therapy and evaluated in relation to their achieved quality at the end of therapy. The "ErgeDoku-B-Form" describes a variety of problem areas as well as questions related to medication and changes in data structure are presented. Results reveal high levels of acceptance in daily clinical practice (indicating clear and appropriate semantics) as well as highly significant relationships between patient and therapist evaluations. PMID- 7480587 TI - [Between isolation and new hope: the psychosocial status of HIV-infected women]. AB - HIV-infected women were an increasingly larger group among the risk groups in Germany. In order to illuminate the psychological situation of HIV-infected women in Germany, 100 women with HIV disease were enrolled in a basic study. 72% of the patients were intravenous drug addicted, 28% were infected through other means. The study population comprised patients of all stages of infection. We found 52% in the first stages, 28% in ARC and 20% with AIDS. Evaluated data included medical, socio-demographic, and data regarding the patients' history of drug abuse. The SCL-90-R, a psychological symptom inventory, was applied. Questions about differences between the two groups of HIV-infected women (iv drug addicted vs non-drug addicted) were answered first. Second, we described three clusters of HIV infected women with regard to different levels of psychological distress. Important factors to distinguish between these clusters are the subjective impact of physical distress (e.g. suffering from HIV-associated symptoms) and loss of joy and interest in sex. In order to illuminate the psychosocial situation, we discuss the specific situation of three patients. PMID- 7480588 TI - [Need for psychosocial counseling--a comparison of women and men in East and West Germany]. AB - The increasing use of psychosocial medical care by women has resulted in a wider range of counselling facilities for specific problems concerning women over the last few years. According to our survey including the population of both Western and Eastern Germany, however, there is no evidence that women have a greater need for counselling activities than men. Needs for counselling are more freely admitted by women than by men; counselling needs for so-called "women's problems" are more commonly observed in Eastern Germany, whereas male oriented problems predominate in Western Germany. The highest need for counselling exists in the field of "Health" and "Social Security". PMID- 7480589 TI - [Analysis of etiologic models of disease development and continuation of bulimia nervosa using structural equation modeling]. AB - In contrast to the great number of etiological models for bulimia nervosa, few conceptions determine the therapeutic practice. Two of these models, the ego psychological model of reduced impulse-control and the behavioural model focussing the influence of "restrained eating" are empirically compared. Both models are not supposed to be of ubiquitous validity, but valid for subgroups of bulimic women. The theoretical assumptions are translated into structural equations and a sample of 127 bulimic women is divided up into 2 subsamples according to criteria which are presumed to define the validity of both etiological models. In agreement with the assumptions the "restrained eating" model shows the better "fit" with the one subsample while the ego-psychological model is more adequate for the other group. PMID- 7480590 TI - [Significance of symbolism in analytic and analytically oriented psychotherapy]. AB - Since the very beginning of psychoanalysis there has been a long lasting discussion about the definition and the role of symbols and symbolization within the theoretical framework of psychoanalysis. The discussion has been summarized by Orban (1976) and Speidel (1978). In this paper a psychoanalytic understanding of creative processes and the relevance of symbolization for the course of treatment is presented. PMID- 7480591 TI - [Using the method of central relationship conflict topic in a study of the process and outcome of long-term inpatient group psychotherapy]. AB - Within a research project dealing with the process and outcome of inpatient group psychotherapy, the core conflictual relationship theme method (CCRT) was used to determine if the central basic interpersonal problems of patients differ in comparison of individual pre- and postreatment sessions, if the CCRT determined at the beginning of treatment is related to outcome, and if there is a similarity between the CCRT from individual and group sessions. Comparisons of the CRRT( components) from the beginning and the end of treatment (related to 19 patients) indicated marked changes on the levels of wishes and responses from others as well as "formal" changes of the narratives (temporal relations as well as the objects mentioned). The second part of the study (n = 26) revealed differences between subgroups determined on the basis of differential outcome measures with respect to the structure of their conflicts. The similarity of the CCRT from group and individual sessions which was studied for nine patients appeared to be moderate. The study revealed that the CCRT might need some minor revisions but appears to be an important method to thoroughly describe the effects of psychotherapies. PMID- 7480592 TI - [The Patient Satisfaction Questionnaire: a suitable tool for quality control in the physician-patient relationship?]. AB - The relationship between doctor and patient receives increasing interest in the public press and in scientific discussions. An increasing number of intervention studies aiming at an enhancement of doctor-patient communication is currently in action. This paper examines the potential of a patient satisfaction questionnaire (PSQ) to assess the need for such an intervention study or to identify those residents who would profit most from an intervention. We investigated a sequential sample of 257 outpatients at the Outpatient Clinic of a Medical University Clinic (137 men, 120 women, mean age 42 +/- 17 years) who were seen by eleven residents for the first time. We used the german translation of an american 14-item questionnaire. Typical psychosocial characteristics of the sample were also assessed. 73.2 percent of the patients are "satisfied" or "very satisfied" with various physician activities during the examination, the mean sum score of the PSQ is 60.1 +/- 7.1 points (range: 39-70 points; maximum score: 70 points). Residents reach a mean total score between 58 and 63 points. Neither in single items nor in the sum score was there any significant difference between residents. We found no differences in PSQ values attributable to sociodemographic variables. The results show that the PSQ is not able to identify those residents to whom intervention studies should be offered in order to enhance psychosocial skills. Its use as an evaluation tool seems also limited because patient satisfaction is very high already prior to an intervention. PMID- 7480593 TI - [Effects of psychological treatment in the general hospital: follow-up study of patients with somatoform behavior]. AB - Over a period of 35 month patients with psychic disorders were treated by a psychological consultation-liaison-service during their stay at a general hospital. A follow-up investigation using a questionnaire was carried out in 1990 in patients with somatization. 66.6% of the patients with somatization took part in the investigation. 47.6% of these patients initiated a psychosomatic treatment after discharge from hospital. 62.8% of the patients described their general state of health as improved. The frequency of symptoms and the amount of restrictions in cause of symptoms could be explained by two factors: Patients without an opportunity to continue a psychosomatic treatment and patients who anticipated negative consequences in case of a psychosomatic treatment reported significantly more complaints and a worse health status. PMID- 7480594 TI - Adult immunisation--too little, too late? PMID- 7480595 TI - Vitamin A deficiency in the South Pacific. AB - Vitamin A deficiency is a major cause of morbidity, mortality and blindness among children. Although vitamin A deficiency is known to affect many children in developing countries, the magnitude of the problem in the South Pacific region is unclear. METHODS: Five cross-sectional surveys for vitamin A deficiency were conducted between 1989 and 1992 in the Republic of Kiribati, Tuvalu, the Republic of Vanuatu, Solomon Islands and the Cook Islands. RESULTS: In total, 10,673 children between the ages of 6 and 72 months were examined for clinical signs of vitamin A deficiency (nightblindness and xerophthalmia). The prevalence of xerophthalmia was 14.76% in the Republic of Kiribati, 1.55% in Solomon Islands, 0.59% in the Cook Islands, 0.28% in Tuvalu, and 0.11% in Vanuatu. The most common clinical findings were Bitot's spots followed by nightblindness. Xerophthalmia were more common among boys (Kiribati P < 0.001, Solomon Islands P = 0.03) and tended to occur in older preschool children (P < 0.0001). CONCLUSIONS: These studies suggest that vitamin A deficiency is a public health problem in the Republic of Kiribati and Solomon Islands. PMID- 7480596 TI - The school health service in Fife: a survey of the views of school head and guidance teachers. AB - A survey of a sample of primary and secondary school headteachers and guidance teachers in Fife was carried out in November 1993. It gathered information about their views on the current school health service. The findings showed that there was a high level of satisfaction with the service but there was inadequate communication between the health service staff and school staff. Teachers clearly indicated a desire to give health education matters a higher priority and issues such as smoking and drug taking by young people in Fife were specifically mentioned. Increased collaboration with health staff in delivering the health and social curriculum appeared to be welcomed. Examples of early implementation of the health-promoting school concept were given. It is important that, at this time of change in school health services, there is adequate discussion with local authority education departments. Improved communication between health services and education staff at all levels will be important to ensure a smooth evolution of school health services and to facilitate the wide introduction and development of the health-promoting school concept. PMID- 7480597 TI - Evaluation of the Fife health visitor proactive training programme. AB - Proactive health visiting refers to a model of health visiting practice which involves actively seeking and meeting the health needs of a defined community. In Fife a working group was established to draw up an operational definition of this concept. Training for health visitors was introduced on a patch-by-patch basis in Fife between December 1992 and May 1993 and an evaluation was subsequently carried out to assess whether or not the proactive concept was being implemented and practised throughout Fife. The survey results suggest that the proactive training has been successful in bringing about a change in health visitor practice over a relatively short time period. With an increasing amount of health visiting work taking place in clinic settings it will be important to define the relative roles of general practitioners, practice nurses and health visitors. A number of operational constraints were identified by the survey and these tended to limit the implementation of the proactive approach. However, we believe that the initiatives which have been successfully introduced should be encouraged and supported, and recommend that this approach be more widely adopted. PMID- 7480598 TI - A corporate needs assessment for the purchase of district nursing: a qualitative approach. AB - A qualitative research methodology was applied to a corporate assessment of need for district nursing services. Semi-structured interviews were held with district nurses and the transcripts analysed for common themes. The major determinants of district nurses' roles were the priorities and practices of other community-based services, particularly social services and primary care. Purchasing and contract setting in this area needs to focus more on the interface with other services. PMID- 7480599 TI - Should nurses in Israeli primary care clinics be expected to manage streptococcal throat infections? AB - INTRODUCTION: Legal regulations in Israel allow nurses to perform only limited clinical procedures. OBJECTIVE: To determine the probability of streptococcal infection in adults with sore throat, as assessed clinically by standard nursing procedures. DESIGN: Using throat culture as the standard, the contribution of various clinical findings (fever, exudate, erythrocyte sedimentation rate (ESR), white blood count) to the determination of the diagnosis of streptococcal infection was assessed, using logistic regression analysis. SETTING: Israeli general practice. PATIENTS: 100 consecutive adult patients presenting with a red, sore throat. RESULTS: Six patients showed the full clinical picture of exudate, increased ESR and leukocytosis, with an 82% probability of streptococcal infection. Forty-nine patients showed none of these three findings, and only one of them (among the three patients with rhinitis) had a positive throat swab. Forty-five patients showed an intermediate clinical picture which did not provide a reliable basis for the diagnosis or exclusion of streptococcal infection. Fever alone did not significantly (P < 0.05) predict streptococcal infection. CONCLUSIONS: Nurses may safely discharge adult patients with a red, sore throat without antibiotic treatment only if they have no additional signs or symptoms. The few patients with all the clinical findings may be treated with antibiotics without a throat swab. All other patients should be referred for examination by a doctor. PMID- 7480600 TI - Access, expectations and communication: Japanese mothers' interaction with GPs in a pilot study in North London. AB - The objectives of this study were: (i) to assess access to British GPs among Japanese mothers in Barnet: (ii) to describe what Japanese mothers expect from a consultation with a GP and; (iii) to understand what the barriers are for Japanese mothers in communicating with GPs. Semi-structured interviews were conducted with eighteen Japanese mothers residing in Barnet, North London, England. The interviews were recorded and their content subsequently transcribed and translated into English. First, gaining quick access to GPs was found to be much more difficult than communicating with GPs. The receptionist and appointment systems were sometimes seen as impediments to early consultation. Second, language was regarded as the biggest barrier in face-to-face communication. In particular the Japanese mothers found it difficult to understand colloquial expressions which were sometimes favoured by GPs. Third, GPs were unfamiliar with the Japanese mothers' expectations and experiences within their own health care system. These findings in relation to access, expectations and communication have provided us with some pointers as to the nature of the current difficulties experienced by one ethnic group. PMID- 7480601 TI - The use of desk-top cholesterol analysers in general practice. AB - An examination of the use of desk-top cholesterol analysers was performed within 10 practices in Kirklees, West Yorkshire. The survey entailed a questionnaire survey of the principal users of desk-top cholesterol analysers and the analysis of two laboratory quality control samples. The majority of practices used Boehringer Lipotrend, with frequency of use varying from < 5 tests/week to a maximum of 20/week. Training by manufacturers was absent in four practices. The maintenance of analysers was inadequate in seven practices as was quality control in five practices. Analysis of laboratory quality control samples showed about 50% of results within 0.5 mmol/l of mean, but 20.6% differed from mean by > 1 mmol/l. The coefficients of variation were 18% and 20% respectively. The use of desk-top cholesterol analysers is increasing in general practice, but improvements in training and quality control are necessary to ensure precise and accurate results with near-patient testing. PMID- 7480602 TI - Evaluation of methods for calculating census health indicators for GP practices. AB - The patients registered with a general practice are usually spread over many census areas and overlap with the distribution of neighbouring practices, so a validated method of aggregating census data to describe the characteristics of practice patients is required. Four methods were used to provide estimates of the percentage of patients aged 75 years and over from census data for 81 practices in Suffolk, England, and these were compared with values derived from the FHSA patient register. Census values for practice areas produced better estimates than those based on the location of the surgery, but the best methods were based on patient-weighted averages of ward and enumeration district data. The finer geographical detail of enumeration districts did not produce substantially more accurate estimates than the ward-level data: both gave estimates with limits of agreement within 2% of the patient register values. Errors in the census, errors in patient registers and selective geographical distributions of practice patients prevent close matching of census and register measures, but two of the methods tested produced estimates that allow broad comparisons between practices. PMID- 7480603 TI - Deprivation and hospital use in a non-metropolitan region of England. AB - OBJECTIVE: To investigate influences on hospital use in a non-metropolitan region of England (Wessex). DESIGN: Least-squares regression analysis was used to identify area predictors, at local authority level, of number of consultant episodes and (separately) of bed use. Variables were assigned for deprivation, strength of primary care, private sector provision, and local levels of ill health. Short-stay (less than 30 days) episodes and bed use were examined separately from long stays. RESULTS: There were substantial differences between the regression models for number of episodes and bed use, and between those for short stays and long stays. Significant predictors of the number of short-stay episodes were: number of general practitioners, standardised mortality ratio and proportion of residents with limiting long-term illness; these variables did not predict bed use in short-stay episodes. None of the variables was significantly associated with long-stay episodes or bed use. CONCLUSIONS: Analysis of geographic variation in hospital use should distinguish between short-stay and long-stay use, and should defend the choice of number of admissions versus bed use as the dependent variable. Deprivation was not shown to be a factor influencing bed use in Wessex. This finding argues against the use of a single funding formula across the whole of the country. PMID- 7480604 TI - A widespread community outbreak of E coli O157 infection in Scotland. AB - Between the end of March and the end of May 1994, 22 cases of laboratory confirmed infection by E coli O157, phage type 4, verotoxin type 1 & 2, were reported to the Scottish Centre for Infection and Environmental Health. Although cases were distributed throughout six health board areas, pulsed field gel electrophoresis indicated that the causative organisms were clonal or very closely related. The eight earliest cases had a connection with a chain of butchers' shops although no organisms were isolated from food or surfaces. A case control study of 9 cases and 27 controls showed a statistically significant association between consumption of burgers and illness, leading to a statement from the Chief Medical Officer emphasising the importance of thorough cooking of burgers and other meat. This outbreak highlights the importance of a unified approach to epidemiological investigation when several administrative areas are involved. PMID- 7480605 TI - Outbreak of Vibrio cholerae 01 in Hong Kong related to contaminated fish tank water. AB - An outbreak of 12 cholera cases, caused by Vibrio cholerae eltor inaba, occurred in Hong Kong during a three week period in June-July 1994. Only adults of both sexes were affected. Epidemiological investigations showed linkage in all cases with consumption of seafood, including shellfish, mantis shrimps and crabs. Microbiological findings demonstrated that contaminated seawater in fish tanks used for keeping alive these seafoods is the most likely vehicle of transmission. Aggressive control measures, promptly instituted, included prohibition of use of contaminated typhoon shelter water in fish tanks, use of seawater with E. coli counts below 610 organisms/100 ml, and the banning of unlicensed food sampans in typhoon shelters. These measures, coupled with public announcements and an active health education campaign on food safety and personal hygiene, abruptly terminated the outbreak. Places which practise the use of seawater, from probable contaminated sources, to keep alive their seafood for human consumption should be alerted to the possibility of transmission of Vibrio cholerae through this route. PMID- 7480606 TI - Dispelling the myth that 50 percent of U.S. schoolchildren have never had a cavity. AB - The erroneous claim that 50 percent of U.S. schoolchildren have never had a cavity has taken on the virtues of truth through frequent and widespread restatement. The 50-percent caries-free statement is an excessively optimistic misrepresentation by the media of the 1986-87 survey of oral health among schoolchildren by the National Institute of Dental Research because it only tells part of the story--it ignores dental disease in the primary dentition. This article documents that numerous public policy papers reflect failure to consider primary tooth caries data. Consequently, a significant disease burden has been overlooked. The article reviews the persistent underreporting of children's caries experience in policy documents and the dental literature, and reviews additional epidemiologic studies of caries reported in U.S. dental literature since 1985. Dental caries remains the single most common disease of childhood that is not self-limiting or amenable to a course of antibiotics. The popular statement that half of U.S. schoolchildren have never experienced tooth decay fails profoundly to reflect the extremity and severity of this still highly prevalent condition of childhood. At a time of extreme pressure on the Medicaid Early and Periodic Screening, Diagnosis, and Treatment budget this uncritically held belief is leading to inappropriate policy and funding decisions that can put the health of children at risk. PMID- 7480607 TI - Reinventing vital statistics. The impact of changes in information technology, welfare policy, and health care. AB - Vital statistics offers a case study in the potential of new information technology and reengineering to achieve better public sector performance. New technology--notably the shift from a paper to an electronic process for recording vital events and transmitting the data to public agencies--is creating opportunities to produce more timely, accurate, and useful information. The furthest advanced innovation is the electronic birth certificate. At the same time, changes in welfare policy and health care--including efforts to establish paternity at the time of birth and to improve health care outcomes--are creating pressures for more policy-relevant data about vital events. In addition, the rise of integrated health plans and health information networks is radically altering the organizational context of vital statistics. On the basis of a State-by-State survey of vital statistics officials, the authors estimate that at the end of 1994, 58 percent of all births in the United States were being recorded on an electronic birth certificate and communicated to a public agency electronically. Nearly all respondents reported that the electronic birth certificate brought improvements in both timeliness and accuracy of data. Achieving the full promise of the new technology, however, will require more fundamental changes in institutions and policies and a reconceptualization of the birth certificate as part of a broader perinatal information system. PMID- 7480608 TI - Interactions between hospital and nursing home use. AB - National estimates are provided, for the first time, of the number of hospitalizations in a year for elderly persons who also experience some nursing home use, and patterns for this interaction are described. In 1987, 816,000 persons were transferred from nursing homes to hospitals, constituting 8.5 percent of all Medicare hospital admissions for persons ages 65 and older. Another 347,000 hospital stays involved people admitted from the community and discharged to a nursing home. The reporting of discharge destination on Medicare hospital bill data in 1987 also is analyzed. It was found that these data may have underreported a nursing home as the destination by between 15 and 20 percent. The magnitude of hospitalizations of nursing home residents suggests that programs aimed at improving nursing home care might have an important impact on total days of hospital care, and that it is important to learn more about the optimal use of expensive hospital care. PMID- 7480609 TI - Our health paradigm in peril. AB - Health care systems are rapidly shifting attention from providing health care to producing health, profoundly altering how and which services are provided. To free up individual and collective resources for investment in activities with a greater impact on health, less care will be given. This paper posits that the current model--increased health resources make for better health care make for better health status--is too simplistic a system. Structural problems inherent in this model are being observed as the boundaries of the paradigm are pushed. Resources are limited, and health outcomes are no longer being improved despite the application of large percentages of Gross National Product. A new health paradigm is emerging, one with increased focus on health prerequisites such as housing, minimum decent income, food, education, and good social and physical environment. PMID- 7480610 TI - Translating research into MCH service: comparison of a pilot project and a large scale resource mothers program. AB - This study examines the process and effect of translating a pilot research project into a large-scale service program. In a pilot resource mothers program for pregnant teenagers, participants had fewer low birth weight infants than teenagers in the comparison group. In the corresponding large-scale service program, a similarly positive effect on low birth weight was not seen. In an effort to understand how these differences occurred, the evaluation methodologies and key characteristics that describe the background, infrastructure, components, and service providers of the two projects were compared. Important differences between the pilot project and the service program were seen in funding stability, diversity of staff, community versus health department ownership of the program, caseloads, and levels of training and supervision. It seems probable that these differences brought about changes in the intensity and character of the intervention from the pilot to the service program, leading to a reduction of the intervention's efficacy in reducing the number of low birth weight infants. The implications of these findings for researchers and program planners are discussed. PMID- 7480613 TI - Racial variation in spontaneous fetal deaths at 20 weeks or older in upstate New York, 1980-86. AB - The distribution of spontaneous fetal deaths (at age 20 weeks or more) by maternal race has received considerably less study than other adverse pregnancy outcomes. The purpose of this study was twofold--(a) to describe spontaneous fetal deaths among white, black, and American Indian women and (b) to determine if there was any variation by International Classification of Diseases, Ninth Revision (ICD-9) cause of death, gestational age at death, or maternal age at loss among these groups of mothers. Using the fetal death certificate registry maintained by the New York State Department of Health, 8,592 spontaneous fetal deaths at age 20 weeks or more were identified among upstate (exclusive of New York City) mothers between 1980 and 1986. By race it was 7,300 for white women, 1,257 for black women, and 27 for American Indian women. Spontaneous fetal death rates varied by maternal race as listed on vital records--black, 13.5 per 1,000 total births, white, 8.3, and American Indian, 8.1. The three leading causes of death (ICD-9,779, 762, and 761) did not vary by maternal race. Gestational age at death, imputed from last menstrual period, did vary by maternal race. Fetal deaths to white and black mothers were observed to occur most often between 24 weeks of pregnancy (39 percent) and 32 weeks (43 percent), while American Indian fetal deaths generally occurred later (more than 33 weeks) in pregnancy (41 percent). Most spontaneous fetal deaths occurred to mothers ages 20-29 regardless of race. Black teenage mothers, however, experienced the largest proportion of losses(23 percent) compared with white (10 percent) and American Indian (I I percent) teenage mothers. PMID- 7480612 TI - Outbreak of Shigella flexneri linked to salad prepared at a central commissary in Michigan. AB - In August 1992, the Michigan Department of Public Health was notified of a cluster of persons with Shigella flexneri infections, all of whom had eaten at different outlets of a single restaurant chain. The chain prepared many foods at a central kitchen. A matched case-control study to determine risk factors for illness among patrons of the restaurant chain was undertaken. An inspection of the commissary and a review of commissary inspection and employee records were conducted. Of the 46 patients identified, 44 had eaten tossed salad versus 33 of the 71 controls (matched odds ratio = 56.9; 95 percent confidence interval 5.0, 648.1). Improper salad preparation techniques were used, and the salad preparation area had not been inspected in several years. Some salad preparers had been ill shortly before the outbreak but continued to work. The greater use of central kitchens could lead to larger outbreaks of illness related to improper food handling. Raw vegetables are a potential vehicle for transmission of shigellosis. Coordination of all agencies responsible for inspecting commissaries should be assured. Assuring restriction of ill food handlers will require management to take an active role in identification of ill employees and in the development of incentives to report illness. PMID- 7480611 TI - Continued adverse trends in coronary heart disease mortality among blacks, 1980 91. AB - An abrupt downturn in mortality rates from coronary heart disease occurred in the United States in the mid-1960s, and for the next decade all four major sex-race groups experienced virtually identical rates of decline. Beginning around the mid 1970s, however, trends for blacks and whites began to diverge, with a deceleration in the annual fall in rates for blacks. The recent release of mortality data extending through 1991, with correction of the denominator estimates in the 1980s using the 1990 census, demonstrate a striking linearity of this trend over the entire decade. In 1989, for the first time since the category of coronary heart disease has been recorded in vital statistics, the age-adjusted death rate for it among black men exceeded that of whites. As a result of the divergent trends among men, an excess of 4,000 deaths of blacks were recorded in 1991 alone. Among women, coronary heart disease mortality was higher among blacks at the beginning of this period, and the average annual percent decline was only two-thirds that of whites. As a result, the absolute mortality gap between blacks and whites steadily increased from 19 to 33 percent (1980 to 1991). This study indicates that the factors that have led to the decline in coronary heart disease have not influenced all demographic groups equally over the last decade. PMID- 7480614 TI - Prevalence and consequences of smoking, alcohol use, and illicit drug use at five worksites. AB - Employers are becoming increasingly concerned about the consequences and costs of substance use in their workplaces. Despite this heightened awareness, little information is available to guide them in setting up worksite-based prevention and assistance programs. Most estimates of the prevalence or consequences of substance use are derived from large national surveys of households or individual persons. The primary contribution of this research to the public health literature is the empirical results from a unique data set. In particular, this study presents results of a survey administered to more than 1,200 employees at five different worksites. Descriptive statistics for the prevalence of smoking, alcohol and illicit drug use, prescription drug misuse, and workplace consequences, such as reduced performance and absenteeism, are reported, as well as findings from a multivariate analysis of substance use prevalence and consequences. Compared with national averages, workers at these five sites tended to have substance use profiles similar to or slightly lower than estimates from large national surveys. The study's estimates may help employers identify the extent of a substance abuse problem in their worksites and specific areas to target for possible intervention. PMID- 7480615 TI - Predictors of maintained high-risk behaviors among impoverished women. AB - The researchers sought to explore and describe the demographic, cognitive, psychosocial, and behavioral factors associated with the continued risky behavior of a convenience sample of homeless and drug-addicted women two to four weeks after they had completed an AIDS education program. The sample included 942 crack users and 767 women who had multiple sex partners. Analyses revealed that impoverished women who maintained multiple sexual partners were less likely to be in drug recovery programs than in homeless shelters. They were more likely to share needles and be involved sexually with male injection drug users compared with impoverished women who did not maintain multiple sexual partners. Persistent crack users were older than those who reported cessation of crack use, were more often African American, and were more likely to have sex partners who were injecting drug users. Women who demonstrated less improvement in depression and distress scores, concerns, use of affective coping, appraisal of threat, and social support were more likely to maintain crack use and multiple partners. The study's implications for the design of intervention programs aimed at risk reduction based on ethnicity are discussed. PMID- 7480616 TI - Multidimensional causal model of dental caries development in low-income preschool children. AB - Despite the decline in the incidence of dental caries in the United States over the past several years, the condition remains a significant problem for the nation's poor children. Efforts to identify the factors responsible for caries development in samples of children of low socioeconomic status have primarily focused on a limited number of variables, and those have been predominantly biological (mutans streptococci, for example). Resulting models of caries development have usually shown good sensitivity but poor specificity. They have had limited implications for treatment. In an effort to produce a comprehensive model of caries development, 184 low-income preschool children were clinically assessed for mutans streptococci and for decayed, missing, or filled surfaces of deciduous teeth twice, first at age 4 years (baseline) and again a year later (year 1 assessment). As the clinical assessments were being done, caretakers were being interviewed to obtain data from five domains: demographics, social status, dental health behaviors, cognitive factors such as self-efficacy (self confidence) and controllability, and perceived life stress. Data were analyzed using a structural equations modeling approach in which variables from all domains, plus baseline decayed missing and filled surfaces and baseline mutants, were used together to create a model of caries development in the year 1 assessment. Results confirmed earlier work that suggested that caries development at a 1-year followup was strongly dependent on earlier caries development. Early caries development in this sample was determined in part by mutans levels and by dental health behaviors. These behaviors themselves were accounted for partly by a cognitive factor. The results support the advantages of employing multidimensional models and provide some direction for intervention to reduce caries incidence. PMID- 7480617 TI - Misunderstanding of 'safer sex' by heterosexually active adults. AB - To assess the understanding of safer sex among heterosexual adults, people enrolled in human immunodeficiency virus (HIV) education trials at a sexually transmitted disease (STD) clinic and a university student health service were surveyed concerning sexual behavior with their latest reported partner. Of 646 sexually active persons enrolled in the trials, 233 (36 percent) reported having had safer sex with their latest partner; 124 of them (53 percent) also reported having vaginal or anal intercourse without a condom during that sexual encounter. Among the 124 who reported safer sex despite having intercourse without a condom, only 23 percent reported asking partners about their HIV status, 46 percent had asked about intravenous drug use, and 47 percent had asked about the number of prior sexual partners. For 34 percent of those surveyed, the length of the sexual relationship with their latest partner was 1 month or less, and 18 percent estimated that this partner had had 11 or more prior sexual partners. STD clinic participants characterized intercourse without a condom as safer sex more often than student health service enrollees (76 percent versus 39 percent, P < 0.001). The concept of safer sex is often misunderstood by persons engaging in behavior at risk for HIV transmission, and the level of misunderstanding differs among samples. Interventions to reduce transmission of HIV must confront misconceptions about the risk of sexual intercourse without condoms and include specific instructions understood by the targeted group. PMID- 7480618 TI - What is the addicts' grapevine when there's 'bad dope'? An investigation in New Jersey. AB - After a rash of fatal overdoses among drug users that was attributed to the synthetic narcotic analgesic fentanyl, the New Jersey Department of Health conducted street interviews with 160 injection drug users in an attempt to identify the channels through which this population had heard about the outbreak and to gauge drug addicts' responses to the incident. The results of the investigation suggest that the drug users learn about such severe threats to health from a variety of sources. The frequency with which some of these sources are reported differs significantly according to the sex of the drug user and, even when sex is controlled, the frequency may vary substantially from city to city in a relatively limited geographic area. Although television was, for this population, a more important source of information about the outbreak than was any other formal means of communication, drug users did not regard TV as a reliable source of good information about "bad dope." Moreover, it does not appear that broadcasts of public warning messages about such substances are a guarantee that addicts will not search for the drug. The data reported in this study point up a need for health officials' greater understanding of the channels through which drug users receive information on threats to their health. The study also provides an understanding of how public health messages are perceived and processed by needle users. The final lesson is the need for close collaboration among drug enforcement personnel, testing laboratories, and health officials in the various affected locales to clarify the public health message. PMID- 7480619 TI - Sustained effects of educating retailers to reduce cigarette sales to minors. AB - Despite state laws prohibiting the purchase of tobacco by minors, the ease with which underage youth can purchase cigarettes has been documented nationwide. The public health community as well as policy makers have called for a combination of retailer education and enforcement of laws prohibiting tobacco sales to minors. Enforcement activity may not be feasible in many communities, however, and an educational intervention may be the only option. This paper reports results of a 6-month followup assessment following a face-to-face education intervention with retailers to reduce cigarettes sales to minors in San Diego County, CA. A control experimental group, pre-post design was employed to study the sustained effects of the program on the illegal sale of cigarettes to minors. A total of 236 stores were visited by minors, ages 14-17 years, with the intent of purchasing cigarettes. Information was collected three times: pre-test, immediately following the intervention, and 6 months after the intervention ended. The groups included a no-treatment control group of 108 stores and an intervention group of 128 that received three educational visits from project staff over a 1-year period. Community education via media and informational presentations was also conducted. As previously reported, a 68-percent pretest sales rate was found for stores overall. Immediately following the intervention, 32 percent of the intervention group and 59 percent of the control group sold cigarettes to minors. These results were maintained 6 months following the conclusion of the intervention. Results are discussed in terms of education versus use of enforcement. PMID- 7480620 TI - The career of Surgeon General Thomas J. Parran, Jr., MD, (1892-1968). PMID- 7480621 TI - An experiment in Tort reform. PMID- 7480622 TI - The Disease Impact Assessment System (DIAS). PMID- 7480623 TI - Analysis of the mechanism of DNA recombination using tangles. PMID- 7480624 TI - Overexpression of integral membrane proteins for structural studies. PMID- 7480626 TI - Space and radiation protection: scientific requirements for space research. AB - Ionizing radiation poses a significant risk to humans living and working in space. The major sources of radiation are solar disturbances and galactic cosmic rays. The components of this radiation are energetic charged particles, protons, as well as fully ionized nuclei of all elements. The biological effects of these particles cannot be extrapolated in a straightforward manner from available data on x-rays and gamma-rays. A radiation protection program that meets the needs of spacefaring nations must have a solid scientific basis, capable not only of predicting biological effects, but also of making reliable estimates of the uncertainty in these predictions. A strategy leading to such predictions is proposed, and scientific requirements arising from this strategy are discussed. PMID- 7480625 TI - Radiation protection in space. AB - The challenge for planning radiation protection in space is to estimate the risk of events of low probability after low levels of irradiation. This work has revealed many gaps in our knowledge that require further study. Despite investigations of several irradiated populations, the atomic-bomb survivors remain the primary basis for estimating the risk of ionizing radiation. Compared with previous estimates, two new independent evaluations of available information indicate a significantly greater risk of stochastic effects of radiation (cancer and genetic effects) by about a factor of three for radiation workers, including space travelers. This paper presents a brief historical perspective of the international effort to assure radiation protection in space. PMID- 7480627 TI - Results of space experiments. AB - Life science research in space was started in Europe with the first Biostack experiment flown onboard Apollo 16 in 1972. Biostack was designed to investigate the biological effects of single heavy ions of cosmic radiation. Among several undertakings towards this goal, the Biostack achieved the highest precision in the determination of the spatial correlation of the observed biological response of single test organisms to the passage of single heavy ions, which is the mandatory requirement. It also provided information on the influence of additional spaceflight factors, such as microgravity, on radiation effects and measurements of the spectrum of charge and energy of the cosmic radiation. The experiment was performed as an international cooperation effort. This report gives a summary of the biological data accumulated in this and the follow-on experiments of the Biostack program. PMID- 7480629 TI - Theoretical nuclear database for high-energy, heavy-ion (HZE) transport. AB - Theoretical methods for estimating high-energy, heavy-ion (HZE) particle absorption and fragmentation cross-sections are described and compared with available experimental data. Differences between theory and experiment range from several percent for absorption cross-sections up to about 25%-50% for fragmentation cross-sections. PMID- 7480628 TI - Initiation-promotion model of tumor prevalence in mice from space radiation exposures. AB - Exposures in space consist of low-level background components from galactic cosmic rays (GCR), occasional intense-energetic solar-particle events, periodic passes through geomagnetic-trapped radiation, and exposure from possible onboard nuclear-propulsion engines. Risk models for astronaut exposure from such diverse components and modalities must be developed to assure adequate protection in future NASA missions. The low-level background exposures (GCR), including relativistic heavy ions (HZE), will be the ultimate limiting factor for astronaut career exposure. We consider herein a two-mutation, initiation-promotion, radiation-carcinogenesis model in mice in which the initiation stage is represented by a linear kinetics model of cellular repair/misrepair, including the track-structure model for heavy ion action cross-sections. The model is validated by comparison with the harderian gland tumor experiments of Alpen et al. for various ion beams. We apply the initiation-promotion model to exposures from galactic cosmic rays, using models of the cosmic-ray environment and heavy ion transport, and consider the effects of the age of the mice prior to and after the exposure and of the length of time in space on predictions of relative risk. Our results indicate that biophysical models of age-dependent radiation hazard will provide a better understanding of GCR risk than models that rely strictly on estimates of the initial slopes of these radiations. PMID- 7480630 TI - A Green's function method for heavy ion beam transport. AB - The use of Green's function has played a fundamental role in transport calculations for high-charge high-energy (HZE) ions. Two recent developments have greatly advanced the practical aspects of implementation of these methods. The first was the formulation of a closed-form solution as a multiple fragmentation perturbation series. The second was the effective summation of the closed-form solution through nonperturbative techniques. The nonperturbative methods have been recently extended to an inhomogeneous, two-layer transport media to simulate the lead scattering foil present in the Lawrence Berkeley Laboratories (LBL) biomedical beam line used for cancer therapy. Such inhomogeneous codes are necessary for astronaut shielding in space. The transport codes utilize the Langley Research Center atomic and nuclear database. Transport code and database evaluation are performed by comparison with experiments performed at the LBL Bevalac facility using 670 A MeV 20Ne and 600 A MeV 56Fe ion beams. The comparison with a time-of-flight and delta E detector measurement for the 20Ne beam and the plastic nuclear track detectors for 56Fe show agreement up to 35% 40% in water and aluminium targets, respectively. PMID- 7480631 TI - Flow-cytometric identification and follow-up of mice exposed to x-irradiation: evaluation of a model system. AB - An experimental model system is presented that allows the identification and follow-up of mice exposed to ionizing radiation using flow-cytometric measurements of peripheral blood cells. In an experiment, properties of peripheral blood cells were analysed with flow cytometry for a rapid identification of individuals exposed to radiation. Individuals were then followed longitudinally in an attempt to identify those developing neoplasias. Male CBA mice, 25 days old, were subjected to fractionated x-irradiation (4 x 1.31 Gy) to induce haematopoietic malignancies. By repeated blood sampling followed by flow cytometry, frequencies of micronucleated erythrocytes and of proliferating nucleated cells were determined. Neoplasias were diagnosed by histopathology. Five days after the end of radiation exposure, increased frequencies of proliferating cells, polychromatic erythrocytes and micronucleated normochromatic erythrocytes clearly distinguished the exposed group from the control group. Increased cell proliferation in peripheral blood cells could be used to identify animals with manifest tumours, although these animals were at a late stage of tumour development. Animals with thymic lymphoma (not generalized) could not be identified with the flow-cytometric parameters used. We consider that this model system has a potential use when a small number of risk individuals need to be identified and monitored within a large population. PMID- 7480632 TI - Sensitization to x-rays by sodium arsenite or heat in normal cells and in cells with an induced tolerance for heat and arsenite. AB - In this study we compared sensitization to x-rays by heat or sodium arsenite and the effect of an induced heat or arsenite resistance on radiosensitization. Treatment of Reuber H35 hepatoma cells with either heat or arsenite causes a dose dependent radiosensitization. Based on a comparison of isosurvival doses for arsenite and heat, arsenite causes a stronger enhancement of the radiosensitivity. Radiosensitization increases exponentially with increasing sensitizer dose. It is gradually lost when the time interval between irradiation and treatment with heat or arsenite increases, depending on the treatment sequence. For x-rays prior to heat, radiosensitization disappears approximately twice as fast as in the reverse case. Arsenite radiosensitization shows approximately the same kinetics for an isoeffective combination, but slightly longer times are needed for the complete clearance of the interaction. As with heat, an exposure to arsenite induces a stress response in cultured cells which results in the development of an increased tolerance towards a second exposure. Heat and arsenite induce self- as well as cross-tolerance. The reduction in arsenite or heat toxicity in tolerant cells is correlated with a reduction in radiosensitization. The mechanisms for heat and arsenite cytotoxicity appear to be different. A combination of non-toxic doses of heat and arsenite has a synergistic effect on the cytotoxicity. One hour incubation with 0.02 mM arsenite at 41 degrees C has the same cytotoxicity as 0.2 mM after 3 h incubation at 37 degrees C, and the amount of radiosensitization induced by these treatments is approximately the same. PMID- 7480633 TI - Adaptive response of the chicken embryo to low doses of x-irradiation. AB - Chicken embryos were x-irradiated in ovo with 5-30 cGy (=priming dose) at the 13th-15th day of development. After 3-48 h, brain- and liver-cell suspensions were x-irradiated in vitro with (challenge) doses of 4-32 Gy. Significantly less radiation damage was observed when the radiation response was measured by scheduled DNA synthesis, nucleoid sedimentation and viscosity of alkaline cell lysates 12-36 h after the priming exposure. In vivo, pre-irradiation with 10 cGy enhanced regeneration as evidenced by the DNA content of chicken embryo brain and liver 24 h following a challenge dose of 4 Gy. From nucleoid sedimentation analyses in brain and liver cells immediately after irradiation with 16 Gy and after a 30-min repair period in the presence of aphidicolin, dideoxythymidine and 3-aminobenzamide or in the absence of these DNA repair inhibitors, it is concluded that a reduction of the initial radiation damage is the dominant mechanism of the "radio-adaptive" response of the chicken embryo. Sedimentation of nucleoids from ethidium bromide (EB) (0.75-400 micrograms/ml)-treated cells suggests a higher tendency of "radio-adapted" cells to undergo positive DNA supercoiling in the presence of high EB concentrations. PMID- 7480634 TI - Induction of gene conversion in yeast cells continuously cultured at high radiation background. AB - The induction of genetic damage was investigated by culturing diploid yeast Saccharomyces cerevisiae D7 cells continuously at radiation levels ranging from 0.383 microSv/h to 1.275 mSv/h by selecting appropriate concentrations of tritiated water in the growth medium. These radiation levels correspond to 3 10,000 times the natural background. Parameters such as growth kinetics, gene conversion frequency at background radiation and after a challenging dose of acute gamma-radiation or alkylating agent N-methyl-N'-nitro-N-nitrosoguanidine (MNNG) were assessed. The gene conversion frequency in most of the assays was in the range of 5-10 convertants per 10(6) cells, as in the case of controls. However, a number of the cultures showed conversion frequencies above 20 per 10(6) viable cells. This stochastic phenomenon occurred more frequently in cells which were incubated at higher radiation levels and for longer durations. This suggests that radiation is responsible for the phenomenon. When subculturing continued beyond 900 h, gene conversion frequencies reverted back to normal values in all cultures in spite of elevated background radiation levels, thus suggesting an adaptive response. The generation time of the cells was 78 min in all cultures irrespective of the radiation level. The response of the cells cultured at elevated background radiation levels to subsequent challenging treatment with gamma-radiation or MNNG was identical to that of the control cultures. Our results suggest that in eukaryotic yeast, low-level radiation may induce an adaptive response to chronic radiation, whereas no such response could be detected when the cells were challenged with acute high-dose exposure or with MNNG. PMID- 7480635 TI - The use of an in vitro technique to predict the absorption of dietary radiocaesium by sheep. AB - The validity of an in vitro extraction technique to predict the availability for absorption of radiocaesium in the sheep gut has been assessed. The technique (a 2 h extraction with caesium chloride) was found to be valid for sources with a low availability for transfer across the gut, but inappropriate for ionic radiocaesium or radiocaesium incorporated internally within herbage. For such radiocaesium sources, which have a high in vitro availability, no correlation was found between in vitro extraction and true absorption measurements. A true absorption value of 0.80 is recommended for these sources, although values for individual sheep in the range 0.60 to 1.00 should be expected. PMID- 7480637 TI - Discovery of X rays. PMID- 7480636 TI - Effects of alpha-particles on survival and chromosomal aberrations in human mammary epithelial cells. AB - We have studied the radiation responses of a human mammary epithelial cell line, H184B5 F5-1 M/10. This cell line was derived from primary mammary cells after treatment with chemicals and heavy ions. The F5-1 M/10 cells are immortal, density-inhibited in growth, and non-tumorigenic in athymic nude mice and represent an in vitro model of the human epithelium for radiation studies. Because epithelial cells are the target of alpha-particles emitted from radon daughters, we concentrated our studies on the efficiency of alpha-particles. Confluent cultures of M/10 cells were exposed to accelerated alpha-particles [beam energy incident at the cell monolayer = 3.85 MeV, incident linear energy transfer (LET) in cell = 109 keV/microns] and, for comparison, to 80 kVp x-rays. The following endpoints were studied: (1) survival, (2) chromosome aberrations at the first postirradiation mitosis, and (3) chromosome alterations at later passages following irradiation. The survival curve was exponential for alpha particles (D0 = 0.73 +/- 0.04 Gy), while a shoulder was observed for x-rays (alpha/beta = 2.9 Gy; D0 = 2.5 Gy, extrapolation number 1.6). The relative biological effectiveness (RBE) of high-LET alpha-particles for human epithelial cell killing was 3.3 at 37% survival. Dose-response curves for the induction of chromosome aberrations were linear for alpha-particles and linearquadratic for x rays. The RBE for the induction of chromosome aberrations varied with the type of aberration scored and was high (about 5) for chromosome breaks and low (about 2) for chromosome exchanges.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7480638 TI - Roentgen. PMID- 7480639 TI - The quest for the laws governing radiations and the search for beneficial innovations. AB - The last few years of the 19th Century saw the initial discoveries of ionizing radiation. These seminal discoveries were followed by an era of intensive studies of the physics related to radiation and clinical applications. The quantum theory revolutionized the ideas about nuclear structure and had a major impact on the physics of radiation. The discoveries of artificial radioactivity and of the neutron and the associated nuclear research have led to the availability of a variety of labeled compounds important to the study of human metabolism. Radioimmunoassay is an example, while other labeled compounds have therapeutic significance. New concepts in the acceleration of electrons and positive ions have contributed to the ability to concentrate radiation energy. New concepts in physics have led to the development of significant and versatile forms of diagnostic imaging. An example is computed tomography. Magnetic resonance is another example of an important physical concept which, decades after its discovery, made possible important applications in imaging and spectroscopy. Unlike ionizing radiation, the less energetic radiofrequency photons can convey information about molecular bonds but do not have sufficient energy to break them. Photon and positron emission scanning and tomography provide external images of internal concentrations of radionuclides, permitting the noninvasive determination of function. Dosimetry is fundamentally important to the diagnostic and therapeutic uses of radiation at both microscopic and macroscopic levels. A variety of radiation measuring instruments have been designed and studied for different purposes, including those based on radiation chemical response. Mammography is just one of the diagnostic applications where dosimetry, detector sensitivity characteristics, the radiation energy spectrum and image resolution are all vital parameters. In radiation treatment, physical and mathematical developments and quantitative radiation biology have increasingly led to optimum conformal radiation treatment, with dosimetry as a guiding parameter. PMID- 7480640 TI - A short history of the radiation chemistry of water. AB - The first hundred years of radiation studies have greatly clarified the chemical processes that are induced by radiation. While the fundamental ionization processes are similar in all systems, properties of the medium such as phase, polarity and composition can greatly affect the chemistry. In all systems, one needs to understand what chemical species are formed, what the internal energies are, how they are dispersed spatially, and what reactions can occur. In this review, the progress that has been made in understanding the chemistry that occurs after a radiolysis event for liquid water is outlined as an example of how progress has been made in radiation chemistry. PMID- 7480641 TI - One hundred years of X rays in biological research. AB - The history of X rays in biological research began almost simultaneously with Roentgen's discovery of his new rays. The history may be unique because of the remarkable collaboration of physicists, chemists, biologists and clinicians- collaborations which have produced and are continuing to produce major contributions to both biological and medical science. These contributions include the use of X rays to investigate molecular structure and function, the first demonstration of induced mutagenesis, the delineation of the cell cycle, the initiation of in vitro and in vivo cloning of mammalian cells, and original studies in DNA repair. The following is a personal overview of the history of some of these developments and their relationship to areas of current biological research. In each case an attempt has been made to trace developments from an early observation or observations to the current day. The history has been divided into two segments, each of approximately 50 years. This division seems appropriate because the separation occurs at approximately the same time as developments which were to play a major role in determining the future of radiation research. PMID- 7480643 TI - Comet assay analysis of repair of DNA strand breaks in normal and deficient human cells exposed to radiations and chemicals. Evidence for a repair pathway specificity of DNA ligation. AB - The induction and resealing of DNA strand breaks in a cell line with a proven defect in DNA ligase I, 46BR, and in two Bloom's syndrome cell lines, YBL6 and GM 1492, were compared to those observed in normal human 1BR/3 fibroblasts after treatment with a variety of genotoxic agents whose lesions are processed by different repair pathways. This analysis was performed using the single-cell gel electrophoresis assay. The three types of cells were found to have similar capabilities to recognize and incise ultraviolet photoproducts and also demonstrated similar amounts of DNA breaks immediately after gamma irradiation. During post-treatment incubation, 46BR cells showed a marked DNA re-ligation defect after ultraviolet radiation damage, GM 1492 cells demonstrated a highly reduced DNA joining ability after relatively high doses of ultraviolet radiation, and YBL6 cells were particularly affected in DNA re-ligation after damage by 4 nitroquinoline-1-oxide. The two Bloom's syndrome cell lines and 46BR cells had a nearly normal ability to reseal breaks resulting from gamma irradiation or treatment with xanthine plus xanthine oxidase. These findings suggest that different DNA ligases may be involved in different DNA repair pathways in human cells. PMID- 7480642 TI - Delayed effects of external radiation exposure: a brief history. AB - Within months of Roentgen's discovery of X rays, severe adverse effects were reported, but not well publicized. As a result, over the next two decades, fluoroscope operators suffered lethal skin carcinomas. Later, case reports appeared concerning leukemia in radiation workers, and infants born with severe mental retardation after their mothers had been given pelvic radiotherapy early in pregnancy. Fluoroscopy and radiotherapy for benign disorders continued to be used with abandon until authoritative reports were published on the adverse effects of ionizing radiation by the U.S. NAS-NRC and the UK MRC in 1956. Meanwhile, exposure to the atomic bombs in Japan had occurred and epidemics of delayed effects began to be recognized among the survivors: cataracts (1949), leukemia (1952) and severe mental retardation among newborn infants after intrauterine exposure (1952). No statistically significant excess of germ-cell genetic effects was detected by six clinical measurements (1956), the F1 mortality (1981), cytogenetic studies (1987) or biochemical genetic studies (1988). Somatic cell effects were revealed by long-lasting chromosomal aberrations in peripheral lymphocytes (1968), and somatic cell mutations were found at the glycophorin A locus in erythrocytes (1992). Molecular biology is a likely focus of new studies based on the function of the gene for ataxia telangiectasia (1995), a disorder in which children have severe, even lethal acute radiation reactions when given conventional doses of radiotherapy for lymphoma, to which they are prone. Also, obligate heterozygote female relatives can be studied for increased susceptibility to radiation-induced breast cancer, as suggested by clinical studies. The tumor registries in Hiroshima and Nagasaki now provide incidence data that show the extent of increases in eight common cancers and no increase in eight others (1994). The possibility of very late effects of A-bomb exposure is suggested by recent reports of increased frequencies of hyperparathyroidism, parathyroid cancers and certain causes of death other than cancer. PMID- 7480645 TI - Kinetics of the formation of chromosome aberrations in X-irradiated human lymphocytes: analysis by premature chromosome condensation with delayed fusion. AB - Human lymphocytes irradiated with graded doses of up to 5 Gy of 150 kV X rays were fused with mitotic CHO cells after delay times ranging from 0 to 14 h after irradiation. The yields of dicentrics seen under PCC conditions, using C-banding for centromere detection, and of excess acentric fragments observed in the PCC experiment were determined by image analysis. At 4 Gy the time course of the yield of dicentrics shows an early plateau for delay times up to 2 h, then an S shaped rise and a final plateau which is reached after a delay time of about 8 to 10 h. Whereas the dose-yield curve measured at zero delay time is strictly linear, the shape of the curve obtained for 8 h delay time is linear-quadratic. The linear yield component, alpha D, is formed entirely in the fast process manifested in the early plateau, while component beta D2 is developed slowly in the subsequent hours. Analysis of the kinetics of the rise of the S-shaped curve for yield as a function of time leads to the postulate of an "intermediate product" of pairwise DNA lesion interaction, still fragile when subjected to the stress of PCC, but gradually processed into a stable dicentric chromosome. It is concluded that the observed difference in the kinetics of the alpha and beta components explains a number of earlier results, especially the disappearance of the beta component at high LET, and opens possibilities for chemical and physical modification of the beta component during the extended formation process after irradiation observed here. PMID- 7480644 TI - DNA sequence analysis of HPRT- mutants induced in human lymphoblastoid cells adapted to ionizing radiation. AB - Radioadaptation to the mutagenic effect of ionizing radiation by pre-exposure of human cells to a low dose has been shown to decrease the proportion of HPRT- mutants of the deletion type. To determine whether point mutations would be affected by the adaptive treatment, the molecular nature of mutations induced after exposure to low, high or low plus high doses was established. DNA sequencing of 38 point mutants which still expressed mRNA was performed using reverse transcription/polymerase chain reaction amplification. Under all conditions, base substitutions were the most common mutational event (range 72 80%), the remainder being frameshift and small deletions. The types and proportions of base changes did not appear to be differentially modified. A clustering of mutations was observed in exon 8, independently of the radiation protocol. About 40% of the mutants exhibited incorrect splicing of mRNA. The lack of striking modifications between the different molecular spectra of point mutations suggests that the low-dose pre-exposure does not affect the production and/or the processing of lesions leading to point mutations. Thus the highly significant effect triggered by the low dose is the preferential reduction of deletion-type mutations. In view of the actual small data set, definitive conclusions will be drawn only when our observations are confirmed or can be generalized to human endogenous loci other than the HPRT locus, which is particularly prone to the recovery of deletion-type mutations. PMID- 7480646 TI - Clastogenic factors in the plasma of Chernobyl accident recovery workers: anticlastogenic effect of Ginkgo biloba extract. AB - Clastogenic factors are found in the plasma of persons irradiated accidentally or therapeutically. They persisted in the plasma of A-bomb survivors over 30 years. Clastogenic factors were found in 33 of 47 Chernobyl accident recovery workers (often referred to as liquidators) in a previous study (I. Emerit et al., J. Cancer Res. Clin. Oncol. 120, 558-561, 1994). In the present study, we show that there is a positive correlation between clastogenic activity and dose and that these biomarkers of oxidative stress can be influenced successfully by appropriate antioxidant treatment. With the authorization of the Armenian Ministry of Health, 30 workers were treated with antioxidants from Ginkgo biloba leaves. The extract EGb 761 containing flavonoids and terpenoids was given at a daily dose of 3 x 40 mg (Tanakan, IPSEN, France) during 2 months. The clastogenic activity of the plasma was reduced to control levels on the first day after the end of the treatment. A 1-year follow-up showed that the benefit of the treatment persisted for at least 7 months. One-third of the workers again had clastogenic factors after 1 year, demonstrating that the process which produced clastogenic factors continued. However, the observation that antioxidants do not have to be given continuously is encouraging for intervention trials on a large-scale basis. These appear justified, since clastogenic factors are thought to be risk factors for the development of late effects of irradiation. PMID- 7480647 TI - Pulmonary carcinogenesis in the F344 and Wistar rat after inhalation of plutonium dioxide. AB - Pulmonary carcinogenesis was compared in female F344 and Wistar rats after inhalation of high-fired 239PuO2. Plutonium particle aggregation, as determined by quantitative light and scanning electron microscopic autoradiography, was greater for the F344 strain than for the Wistar strain. The median survival times were similar in control and low-dose (0.8-1.0 Gy) groups of both strains, but were significantly decreased in the high-dose (34-37 Gy) groups of both strains. Squamous metaplasia was not found in control or low-dose groups of either strain, but was found in 62-65% of high-dose groups of both strains. Adenomatous metaplasia was considerably higher in control and low-dose groups of F344 rats than in Wistar rats. A total of 87 lung tumors were found in 140 exposed F344 rats and 46 lung tumors in 176 exposed Wistar rats. The incidence of lung tumors in F344 rats was 1.7% in controls, 20% in the low-dose group and 82% in the high dose group. The incidence of lung tumors in Wistar rats was 0.1% in controls, nil in the low-dose group and 68% in the high-dose group. About half of all lung tumors in both strains were considered to be the primary cause of death. The median survival times of rats of both strains in the high-dose groups that died with lung tumors were greater compared with rats in these groups that died without lung tumors. In contrast, these differences did not occur among rats in the low-dose groups. The absolute risk was 1900 lung tumors per 10(4) Rat-Gy for F344 rats receiving low doses and nil for Wistar rats receiving low doses, but about 210 lung tumors per 10(4) Rat-Gy for high-dose groups of both strains. The adenomatous tumor phenotype predominated in the F344 strain, while the squamous tumor phenotype predominated in the Wistar strain. Risk of squamous tumors was similar for both strains. Overall, the F344 strain appears to be more "sensitive" than the Wistar strain to formation of lung tumors at low to moderate doses from inhaled 239PuO2 due mostly to an increased incidence of adenomatous phenotype tumors. PMID- 7480648 TI - Prevalence of hepatitis B surface antigen, hepatitis B e antigen and antibody, and antigen subtypes in atomic bomb survivors. AB - On the basis of previous studies showing an association between hepatitis B surface antigen (HBsAg) positivity and radiation exposure in atomic bomb (A-bomb) survivors, we investigated further the active state of hepatitis B virus (HBV) infection by incorporating tests for hepatitis B e antigen (HBeAg) and hepatitis B e antibody (anti-HBe) and HBsAg subtypes into our biennial health examinations. Among 6548 A-bomb survivors for whom HBsAg was assayed between July 1979 and July 1981, 129 persons were HBsAg positive. HBeAg and anti-HBe were measured in 104 of these persons and subtypes of HBsAg in 98 persons. Among those exposed to radiation (average liver dose 0.58 Sv), the odds ratio of HBsAg positivity tended to increase with radiation dose (P for trend = 0.024). The P values for association between the prevalence of HB e antigen and radiation dose and between the prevalence of anti-HBe and radiation dose were 0.094 and 0.17, respectively. The HB antigen subtype adr was predominant over other subtypes in both Hiroshima and Nagasaki, but the distribution of subtypes did not seem to differ in relation to radiation dose. These results suggested that A-bomb survivors remain in an active state of HBV infection and that the mechanism(s) of seroconversion may be impaired. PMID- 7480649 TI - The pO2 in a murine tumor after irradiation: an in vivo electron paramagnetic resonance oximetry study. AB - Using electron paramagnetic resonance (EPR) oximetry with the oxygen-sensitive paramagnetic material, fusinite, we have measured the partial pressure of oxygen (pO2) in the mouse mammary adenocarcinoma MTG-B. The average pO2 in untreated tumors was low (about 5 mm Hg) and decreased with tumor growth. Magnetic resonance imaging and histological examination were used to localize the position of the fusinite with respect to tumor margins and vascularization. The pO2 was generally higher in the periphery than in the center of the tumors, but there was considerable variation among tumors both during normal growth and after radiation treatment. After a single 20-Gy dose, a characteristic pattern of change in tumor pO2 was observed. In irradiated tumors, there was an initial reduction in pO2 (minimum occurred 6 h postirradiation) which was followed by a transient increase in pO2 to levels higher than the preirradiation pO2 (maximum occurred 48 h postirradiation). This work demonstrates postirradiation changes in pO2 of potential radiobiological significance. Compared to other oxygen assessment techniques, EPR oximetry is very useful because it can assess pO2 in the same region of the tumor over the course of tumor growth and during response to treatment. Thus EPR could be used to identify potentially radioresistant tumors as well as to identify tumors with slow reoxygenation. PMID- 7480650 TI - Enhanced radiation-induced cell killing by carboplatin in cells of repair proficient and repair-deficient cell lines. AB - The objective of this study was to determine whether a deficiency for either one of two repair processes influences the phenomenon of enhancement of radiation induced cell killing by carboplatin which has been reported previously in one cell line (V79) and which is presumably a result of an interaction between these two therapeutic modalities. Cell killing was enhanced in cells of four cell lines when the cells were exposed to carboplatin before and during irradiation in either air or hypoxia. In cell lines proficient in both excision repair and DNA double-strand break repair (K1 and AA8), and in a cell line deficient in nucleotide excision repair (UV41), the enhancement was characterized as both a reduction in the shoulder region of the survival curves indicated by a reduced Dq and a reduction in D0 in the terminal region of the survival curves determined for cells exposed in air and under hypoxic conditions. Only the latter effect was observed in a cell line deficient in DNA double-strand break repair (xrs-5). The survival curves were fitted to the data using the repair saturation model and a computer program developed by N. Albright (Radiat. Res. 118, 112-130, 1989). In hypoxia, the reductions in Dq were as great as from 7.0 Gy to 2.1 Gy, 3.3 Gy to 0 Gy and 1.7 Gy to 0 Gy for K1, AA8 and UV41 cells, respectively. Sensitizer enhancement ratios ranged from 1.3 to 1.7 and were similar for irradiation in air and under hypoxic conditions. This enhanced cell killing by carboplatin combined with radiation required levels of the drug sufficient to produce cytotoxicity by the drug alone as exemplified by the UV41 cell line, which is intrinsically sensitive to carboplatin and in which 1/30 of the drug concentration required for the other cell lines produced an enhanced cell killing at an equitoxic dose of only 5 microM. PMID- 7480651 TI - Long-term effects of prenatal exposure to low levels of gamma rays on open-field activity in male mice. AB - The open-field activity of first-generation (F1) hybrid male C57BL/6 x C3H mice irradiated with gamma rays on day 14 of gestation was studied at the following ages: 6-7 months (young), 12-13 months (adult) and 19-20 months (old). Doses were 0.5 Gy or 1.0 Gy. Open-field activity was recorded with a camera. The camera output signal was recorded every second through an A/D converter to a personal computer. The field was divided into 25 8-cm2 units. All recordings were continuous for 60 min. The walking speed of the 1.0-Gy group recorded at 19-20 months was higher than that for the comparably aged control group. The time which the irradiated group, recorded at 19-20 months, spent in the corner fields was high in comparison with the control group at the same age. Conversely, the time spent by the irradiated group in the middle fields when recorded at 19-20 months was shorter than in the comparably aged control group. No effect of radiation was shown for any of the behaviors observed and recorded at 6-7 and 12-13 months. The results demonstrate that such exposure to gamma rays on day 14 of gestation results in behavioral changes which occur at 19-20 months but not at 6-7 or 12-13 months. PMID- 7480652 TI - Theoretical calculation of electronic stopping power of water vapor by proton impact. AB - The energy loss of proton beams in water vapor is analyzed with a full quantum mechanical treatment, the distorted-wave model. This model takes into account distortion effects due to the long-range Coulomb potential. Projectile energies from 10 keV up to 1 MeV are considered. Mean stopping power and equilibrium charge-state fractions are calculated and compared with experimental data. The validity of Bragg's additivity rule is investigated. PMID- 7480653 TI - Clinical image evaluation. AB - The radiologist interpreting mammograms is responsible for an ongoing evaluation of the quality of the initial images produced by the facility. The Mammography Quality Standards act requires an external review of clinical images at least every 3 years from each facility accredited. Clinical image evaluation includes an assessment of position and compression, labeling, image quality, and artifacts. For each of these assessment categories, specific criteria have been identified for distinguishing properly performed from deficient clinical images. PMID- 7480654 TI - Quality control in mammography. AB - This article describes the essential elements of an effective quality control program in mammography. Quality Control tests recently updated in the 1994 ACR Mammography Quality Control Manual, such as the darkroom fog and limiting spatial resolution tests, are described in detail. The article goes on to describe some common problems in mammography quality control, including processor sensitometry and image quality problems. PMID- 7480657 TI - The development and meaning of appropriateness guidelines. AB - Practice guidelines are important for high quality and economical patient care. As gatekeepers to health care, primary care physicians should refer to imaging guidelines when treating a patient with breast disease. The American College of Radiology (ACR) has undertaken the task of publishing guidelines for breast imaging in the hopes of avoiding a plethora of competing, possibly contradictory guidelines written by groups less qualified in imaging protocol than the ACR and with the goal of minimizing the dilemmas of interpretation and patient management often present in breast imaging. PMID- 7480655 TI - The Mammography Quality Standards Act of 1992. History and philosophy. AB - The Mammography Quality Standards Act (MQSA) of 1992 established a precedent in the practice of mammography by creating federal quality standards for all parts of the mammography system. Heralded by some as crucial so all US women can reap the full benefits of early detection of breast cancer, MQSA implementation was delegated to the Food and Drug Administration (FDA) on June 2, 1993. In this article the major historical forces that surrounded MQSA's enactment as well as the FDA's philosophy on regulation development, inspections, and compliance under MQSA is summarized. PMID- 7480658 TI - Breast masses. Appropriate evaluation. AB - Detection and management of a breast mass requires an optimal environment for interpretation, relevant use of clinical information, technically excellent imaging procedures, proper interpretation of the findings, and pertinent recommendations. Masses suspicious for malignancy by only a single modality ideally should undergo some form of biopsy, as the consequences of delayed diagnosis of breast cancer can be devastating. Early invasive breast cancer can be identified and treated if the appropriate evaluation is performed. PMID- 7480656 TI - Pathology of benign and malignant breast disorders. AB - An understanding of the basic anatomy and pathology of the breast is necessary for optimal interpretation of breast imaging studies. This article discusses the normal anatomy and the pathology of benign, high-risk, and malignant lesions of the breast. PMID- 7480659 TI - Evaluation of breast microcalcifications. AB - As screening mammography has become more widespread, the urgency to provide more sensitive and more specific interpretations has also increased. Although many calcifications detected on mammograms are characteristically benign and need no further evaluation, there are many microcalcifications that warrant further evaluation with special views. The importance of their detection lies in the fact that many malignancies are mammographically manifested solely as microcalcifications. Once detected, radiologists have attempted to characterize and stratify microcalcifications by their level of suspicion to improve the predictive value of biopsy recommendations as depicted in Table 1. This is an important endeavor because the cost to society, especially in this era of limited resources, and the collective anxiety produced by benign breast biopsies are both great. When a biopsy is recommended, the breast team, which includes the surgeon, radiologist, and pathologist should ensure that histopathologic correlation is achieved. By participating in this process, keeping track of the results of biopsies generated from our practices, and learning from our colleagues, we can fine tune our interpretive skills. Communication among us and our colleagues from other specialties should be encouraged and can be fostered by the use of the BI RADS lexicon. PMID- 7480660 TI - Management of probably benign breast lesions. AB - Several large-scale prospective clinical studies establish the validity of managing probably benign breast lesions with periodic mammographic surveillance as a safe and effective alternative to immediate tissue diagnosis. This approach to mammographic interpretation is now very widely accepted; the American College of Radiology indeed includes "probably benign - short interval follow-up suggested" as one of the five final assessment categories in the Breast Imaging Reporting and Data System (BI-RADS), which all American radiologists are encouraged to use. There is also general consensus that probably benign interpretations should involve (1) cases restricted to nonpalpable lesions; (2) use of the specific imaging criteria described in the prospective clinical studies; and (3) pre-interpretation comparison with prior mammograms, if available, to ensure that new or progressing lesions undergo prompt biopsy (it makes no sense to recommend follow-up for a lesion that has already demonstrated interval progression when the very demonstration of progression during surveillance is what prompts biopsy instead of continued follow-up). As yet unresolved issues concerning probably benign lesions include whether initial full problem-solving imaging should be performed in all cases, whether to use patient age and lesion size (for solitary masses) as additional criteria in deciding between the management alternatives of mammographic follow-up and immediate tissue diagnosis, and what should be the specific timing, frequency, and duration of the follow-up examinations that constitute the surveillance protocol. PMID- 7480662 TI - Evaluation of the breast undergoing lumpectomy and radiation therapy. AB - As larger numbers of women undergo breast-conserving treatment, it is important for radiologists to understand the usual alterations in the mammogram that result from surgery and radiation and to be able to differentiate these from findings that may indicate a new breast cancer. Scar formation, skin changes, increased breast density, and coarse calcifications are normal findings after breast conserving treatment. Microcalcifications and new masses may indicate recurrent tumor. Fat necrosis and fibrosis can produce changes that may be identical to recurrent cancer. Appropriate follow-up with mammography should help in the early diagnosis of recurrent breast cancer. PMID- 7480663 TI - The current role of ultrasonography in breast imaging. AB - When properly performed and interpreted, US is an indispensable tool in breast imaging. Despite improvements in equipment, the major roles for US in the breast remain the differentiation of cysts from solid masses and the guidance of interventional procedures. Neither of these are trivial tasks, for they have significantly reduced the number of surgical biopsies performed in the United States. Research studies investigating feature analysis and Doppler show future potential for US to be used to differentiate benign from malignant solid masses. PMID- 7480664 TI - Needle biopsy techniques. AB - With the ability to perform good diagnostic mammography, breast ultrasound, ductography, cyst aspiration, abscess drainage, and stereotactic or ultrasound guided percutaneous biopsy, the modern breast radiologist should play the central role in breast diagnosis. The definitive diagnoses afforded by percutaneous breast biopsy that obviate surgery entirely in benign lesions and streamline the therapeutic surgery for malignant lesions now can be obtained routinely. The future holds even more exciting challenges for the radiologist as percutaneous lumpectomy becomes a reality. Thus the breast radiologist, armed with the technology and techniques of the twenty-first century, truly stands on the threshold of a new era. PMID- 7480661 TI - Evaluation of the augmented breast. AB - Silicone gel implants and their complications constitute a public health problem. Millions of American women have these implants and many are concerned about their possible health effects. The history of breast augmentation as well as current concepts in imaging augmented patients are discussed in this article. Special attention is paid to the diagnosis of implant rupture by mammography, sonography, and MR imaging. PMID- 7480665 TI - MR imaging of the breast for cancer evaluation. Current status and future directions. AB - This article discusses the current status of clinical feasibility testing and outlines some of the advantages and disadvantages of contrast-enhanced breast MR imaging. A review of imaging parameters includes the dose of paramagnetic contrast agents, two- versus three-dimensional scanning, and postprocessing data analysis with an emphasis on areas for further development. Principles of interpretation for normal tissue, benign lesions, and invasive cancers are summarized including causes of false-positive and false-negative MR imaging interpretations. Future clinical roles for breast MR imaging such as MR imaging directed biopsies and therapies and MR as a complementary technique to film screen mammography for selected clinical situations are suggested. PMID- 7480666 TI - Digital mammography, computer-aided diagnosis, and telemammography. AB - Film-screen x-ray mammography is an effective tool for the detection and diagnosis of breast cancer. Several limitations of current mammographic technology have been identified, and there is now good evidence that improved image quality and possibly reduced radiation dose could be achieved if mammograms are acquired directly in digital form. In this article, the rationale for digital mammography is presented and various approaches to the design of a digital mammography system are described. Current challenges in developing a practical, clinically acceptable digital mammography system are discussed and applications of digital mammography to computer-aided diagnosis, telemammography, and quantitative imaging are suggested. PMID- 7480667 TI - Communication issues in breast imaging. AB - It is hoped that this discussion makes it clear that unambiguous communication to the woman and health care provider is extremely desirable. This communication along established guidelines using standardized terminology should diminish lack of appropriate follow-up with its inherent negative outcomes. PMID- 7480668 TI - A national breast cancer database. AB - The medical audit of technical and interpretive aspects of mammography has been recommended by a number of expert panels and as part of some regulations that govern the oversight of mammography. The value of the medical audit is to aid practitioners in the performance evaluation of mammography in their facilities. Collection and analysis of audit data at a regional or national level have the potential to allow for the comparison of practice-level performance with regional and national-level performance and to provide a public health tool for the evaluation and improvement of breast cancer detection. In addition, a regional and national-level breast cancer database has the potential for allowing monitoring and tracking of women participating in a screening program. Prior to establishing such a database, data must be protected from discovery and disclosure, and patient and physician confidentiality must be ensured. PMID- 7480669 TI - Why question screening mammography for women in their forties? AB - Groups that do not recommend mammography for all women in their forties do so with reason. Even after seven randomized controlled trials and participation by over 170,000 women it is not clear whether screening women in this age group for breast cancer saves lives. It is clear that in contrast to older women, in women in their forties there is no effect for several years after screening begins, and an uncertain marginal effect after a decade. If an effect is present it may be owing to the fact that women move into an age group when mammography is effective. The uncertainty about mammography in younger women is strong enough that investigators from the seven trials are working to conduct overview analyses of the data from all the trials to assess better at what age breast cancer screening saves lives. There also is enough uncertainty that a trial with almost 200,000 women has begun in the United Kingdom and another one is being considered for Europe. If the answer about mammography for women in their forties is clear, why would such large new trials be started? Meanwhile, evidence exists that breast cancer screening costs a great deal, not just in dollars but in human terms. There also is increasing concern about the possibility of overdiagnosis. The financial costs primarily are owing to charges for screening mammograms. The human costs primarily are owing to false-positive readings and, if present, overdiagnosis. Ultimately, the decision to screen women in their forties for breast cancer involves a weighting of benefits (in mortality reduction) and costs (in dollars, anxiety over false-positive results, biopsy and surgery for noncancerous lesions, and overdiagnosis). Most emphasis in the debate about breast cancer screening has been on the evidence about effectiveness. But the costs are substantial. Groups that do not recommend routine screening mammography for all women in their forties require a greater standard of evidence for effectiveness of screening than those that do. Why is this so? First, there are financial considerations. When a health care intervention is recommended for every person in our society, costs mount fast. At any time, but particularly in an era of cost constraints, it is not only reasonable but incumbent on all involved in health care to look carefully at how much benefit there is for the expenditure and how strong is the scientific evidence. Second, many persons who consider prevention activities argue that there is an ethical reason.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 7480670 TI - Mammography screening and the controversy concerning women aged 40 to 49. AB - The screening controversy arose from inappropriate and scientifically insupportable data analysis of the early results from the randomized, controlled trials of screening. Data were used that were inadequate for making medical recommendations by using subgroup analyses from trials that were not designed to permit subgroup analysis by age. Contrary to suggestions made by some opposed to screening women younger than the age of 50, there are no parameters such as breast density, cancer detection rate, or positive predictive value that change abruptly at age 50 or any other age. The most recent meta-analysis provides statistically significant "proof" that screening can reduce the death rate from breast cancer by at least 24% for women aged 40 to 49. PMID- 7480671 TI - [Magnetic resonance tomography: new results of contrast media research. 8th contrast media expert discussion, 1993]. PMID- 7480672 TI - Hepatic metastases: basic principles and implications for radiologists. PMID- 7480673 TI - Aberrant venous drainage to the liver: imaging implications. PMID- 7480674 TI - SPECT with 2-[fluorine-18]fluoro-2-deoxy-D-glucose (FDG). PMID- 7480675 TI - Aberrant gastric venous drainage in cirrhotic livers: imaging findings in focal areas of liver parenchyma. AB - PURPOSE: To analyze imaging finding in focal areas of liver parenchyma with aberrant gastric venous drainage (AGVD) in cirrhotic livers. MATERIALS AND METHODS: Eighteen patients with AGVD confirmed with computed tomography (CT) during arterial portography and arteriography underwent ultrasound (US) (n = 18), CT (n = 18), magnetic resonance (MR) imaging (n = 12), dynamic CT (n = 14), and dynamic MR imaging (n = 8). Imaging and histologic (n = 2) findings were analyzed. RESULTS: The focal area with AGVD was hypoechoic on US scans (n = 7), hypoattenuating on CT scans (n = 4), hyperintense on T1-weighted MR images (n = 6), and hypointense on T2-weighted MR images (n = 5). Early enhancement was seen with dynamic CT (n = 8) and with dynamic MR imaging (n = 6), without definite staining on arterial phase of arteriography. Histologic findings revealed hyperplastic changes of liver. CONCLUSION: The imaging findings, except for early enhancement, were very similar to those of adenomatous hyperplasia or well differentiated hepatocellular carcinoma. The early enhancement was considered due to early venous return compared with that of the surrounding liver. PMID- 7480676 TI - Effect of temporary occlusion of the hepatic vein on dual blood in the liver: evaluation with spiral CT. AB - PURPOSE: To evaluate the contribution of hepatic veins to the dual blood supply in the liver under temporary hepatic venous occlusion. MATERIALS AND METHODS: Selected hepatic veins in 23 patients with liver tumors were temporarily occluded with a balloon catheter. Computed tomography (CT) arteriography, CT during arterial portography (CTAP), or both were performed with a spiral technique with and without temporary occlusion of a hepatic vein. RESULTS: After hepatic vein occlusion, a well-demarcated, wedge-shaped area of hypoattenuation was seen at CTAP and/or hyperattenuation was seen at CT arteriography in the following regions: left lobe (left hepatic vein), ventral part of the anterior segment and the medial segment except for the ventromedial part (middle hepatic vein), dorsal part of the anterior segment and the ventral part of the posterior segment (right hepatic vein), and dorsocaudal part of the right lobe (inferior right hepatic veins). CONCLUSION: After hepatic venous occlusion, the portal veins become draining veins and the occluded area is supplied with arterial blood alone. PMID- 7480677 TI - Helical CT of the liver: value of an early hepatic arterial phase. AB - PURPOSE: To prospectively evaluate a dual-phase helical acquisition in computed tomography (CT) of the liver. MATERIALS AND METHODS: Two helical CT acquisitions were performed in 103 patients: hepatic arterial phase (HAP) 15 seconds after injection of 2 mL/kg contrast material at a rate of 6 mL/sec and the portal venous phase (PVP) 90 seconds after injection. Detection of focal liver lesions, vascular anatomic features, and perfusion abnormalities were assessed by two reviewers. Liver enhancement was also calculated on every section. RESULTS: Of 119 detected focal liver lesions, nine were seen during HAP only and 40 during PVP only. According to a 0-3 score, visualization of the arterial anatomic landmarks was better during HAP (P < .0001). Five right replaced and two left replaced hepatic arteries were visualized only during HAP. Parenchymal perfusion abnormalities were found in 32 patients during HAP and in four patients during PVP. CONCLUSION: A dual-phase acquisition depicted 8% additional focal liver lesions and outlined the entire vasculature of the liver. PMID- 7480678 TI - Hepatic contrast enhancement at CT: influence of intravenous glucagon in a canine model. AB - PURPOSE: To investigate the effect of glucagon on the timing and degree of hepatic enhancement at computed tomography. MATERIALS AND METHODS: Each of 11 dogs underwent injection of contrast material at two uniphasic rates (0.5 mL/sec and 1.5 mL/sec) with without previous intravenous administration of 0.5 mg of glucagon. Scans were obtained at a single intrahepatic level every 3.5 seconds for 120-160 seconds. Attenuation values in the liver, aorta, portal vein, and inferior vena cava (IVC) were measured. RESULTS: Glucagon administration was associated with greater hepatic enhancement throughout the study: Peak hepatic enhancement was greater at contrast injection rates of 0.5 mL/sec (P = .021) and 1.5 mL/sec (P = .0001). Peak hepatic enhancement also occurred earlier during the glucagon runs. Portal vein enhancement was greater during the glucagon runs at an injection rate of 1.5 mL/sec (P = .032). IVC enhancement was greater during the nonglucagon runs at 0.5 mL/sec (P = .013) and at 1.5 mL/sec (P = .005). CONCLUSION: Intravenous administration of glucagon before contrast material injection produces greater and more rapid hepatic enhancement in a canine model. PMID- 7480679 TI - Idiopathic inflammatory bowel disease: endoscopic-radiologic correlation. AB - PURPOSE: To assess the comparative and adjunctive diagnostic value of double contrast barium enema examination (DCBE) and colonoscopy in cases of idiopathic inflammatory bowel disease. MATERIALS AND METHODS: Radiographic and endoscopic findings were correlated, with use of a standardized evaluation sheet, in a prospective, blinded study in 99 patients with previously well-established Crohn disease of the colon (CD [ n = 51]) or ulcerative colitis (UC [n = 48]). RESULTS: Overall, substantial to almost perfect agreement was found in more than 80% of scores with respect to the items of the standardized evaluation sheet in the colon segments depicted with both imaging modalities. Almost perfect agreement (kappa > 60%) was found for two essential diagnostic and differential diagnostic features: aphthoid erosions and fine granularity. CONCLUSION: DCBE and colonoscopy are complementary imaging modalities for optimal detection of all mucosal and structural colon lesions, except inflammatory lesions without distortion of the mucosal relief and small numbers of inflamed superficial erosions and ulcers. PMID- 7480680 TI - Small hepatocellular carcinoma in patients undergoing liver transplantation: detection with CT after injection of iodized oil. AB - PURPOSE: To assess computed tomography (CT) with iodized oil for depiction of small hepatocellular carcinoma (HCC) before liver transplantation. MATERIALS AND METHODS: Thirty-five consecutive cirrhotic patients underwent CT with iodized oil to determine the presence, number, size, and location of possible nodules. All patients underwent liver transplantation within 4 months after CT. Explanted livers were cut in 8-mm slices that corresponded to axial CT scan planes. Comparison between CT staging and pathologic findings was made. RESULTS: Pathologic studies showed 17 HCC nodules (diameter, 0.9-4.0 cm) in nine of the 35 livers. CT depicted nine of these 17 nodules. Lesion-by-lesion analysis revealed a sensitivity of 53%; CT falsely depicted three additional nodules not confirmed with pathologic findings. Patient-by-patient analysis revealed an 89% sensitivity and an 88% specificity. CONCLUSION: CT with iodized oil, when assessed lesion by lesion, has a low sensitivity. These results must be considered when liver resection is proposed for HCC. PMID- 7480681 TI - Potentially resectable pancreatic adenocarcinoma: spiral CT assessment with surgical and pathologic correlation. AB - PURPOSE: To evaluate the accuracy of spiral computed tomography (CT) in assessing the resectability of small pancreatic ductal adenocarcinoma and to correlate the CT findings with histopathologic and surgical findings. MATERIALS AND METHODS: Spiral CT scans obtained in 64 patients who underwent surgery for potentially resectable pancreatic adenocarcinoma were prospectively assessed for tumor resectability. CT findings were correlated with surgically assessed extent of tumor and pathologic findings. RESULTS: Fifty-seven (89%) of 64 carcinomas were detected with spiral CT. Twenty-four carcinomas were resectable at surgery and 40 were not. The average size of resectable tumors was 3.1 cm (range, 1.0-7.5 cm). The overall accuracy of spiral CT for assessing resectability was 70%. Of resected tumors, 14 were hypoattenuating compared with the remaining pancreas and 10 were isoattenuating. Eleven tumors showed neointimal proliferation in arterioles at histologic examination. CONCLUSION: Further progress in preoperative staging of pancreatic ductal adenocarcinoma with spiral CT should be directed toward improving detection of small pancreatic tumors and assessment of early metastatic disease. PMID- 7480682 TI - Suspect breast lesions: findings at dynamic gadolinium-enhanced MR imaging correlated with mammographic and pathologic features. AB - PURPOSE: To prospectively correlate dynamic contrast enhancement at magnetic resonance (MR) imaging with mammographic and pathologic features of suspect breast lesions. MATERIALS AND METHODS: Forty-nine patients with 51 breast lesions underwent gadolinium-enhanced spoiled gradient-recalled echo (SPGR) MR imaging at 1.5 T, as well as excisional biopsy or cyst aspiration. RESULTS: Twenty-two of 22 (100%) invasive carcinomas 8 mm or more in diameter, including three (12%) not evident on dense mammograms, enhanced 2.0 or more times the unenhanced intensity. One of three predominantly ductal carcinomas in situ and 10 of 26 (38%) benign lesions enhanced 2.0 or more times. Time-intensity curves were not statistically significantly different among enhancing carcinomas, fibroadenomas, or other benign lesions and showed no statistically significant correlations with pathologic size, nodal status, or hormone receptor status of invasive carcinomas. CONCLUSION: MR imaging enhancement of 2.0 or more times had high sensitivity (100%) for invasive carcinomas 8 mm or more in diameter, with moderate specificity (65%). Time-intensity curves showed no significant difference between enhancement of benign and malignant lesions. PMID- 7480683 TI - Pulmonary nodules: improved detection with vascular segmentation and extraction with spiral CT. Work in progress. AB - PURPOSE: To determine whether extraction of pulmonary vessels from computed tomographic (CT) images with automated segmentation improves the detection of pulmonary nodules. MATERIALS AND METHODS: Simulated nodules were superimposed on normal spiral CT images. Eight patients referred for CT assessment of pulmonary nodules were selected for clinical evaluation. Vessels were extracted from both the simulation and clinical study with a three dimensional seeded region-growing algorithm. Three experienced radiologists were asked to locate the nodules and assign a level of confidence to their findings. Sensitivity and proportion of false-positive results per case (FPC) were calculated. Observer performance was evaluated by alternate free-response receiver operating characteristic analysis. RESULTS: Extraction of vascular structures from CT scans improved sensitivity from 63% to 84% in the simulation study and from 58% to 78% in the clinical study. The proportion of FPC decreased from 52% to 24% and from 55% to 12%, respectively. Radiologists performed consistently better with the segmented images than with the original images in both the simulation (P = .006) and the clinical (P = .0013) study. CONCLUSION: Automated vessel subtraction and extraction improves detection of pulmonary nodules. PMID- 7480684 TI - Uncommon pneumoconioses: CT and pathologic findings. AB - PURPOSE: To correlate the computed tomographic (CT) features of pneumoconioses with histologic findings. MATERIALS AND METHODS: Thin-section CT scans obtained in 48 patients with a history of occupational exposure to dust and radiographic changes suggestive of pneumoconiosis were retrospectively reviewed. Histologic samples were available in 22 cases. RESULTS: The most common CT features were as follows: in 21 arc welders, ill-defined micronodules concentrated in the centrilobular regions (n = 15); in 19 graphite workers, small nodular hyperattenuating areas (n = 17) (ill defined or well defined, corresponding to macular lesions along the walls of bronchioles and nodules, respectively), interlobular septal thickening (n = 11), and large hyperattenuating areas (n = 10); in aluminum pneumoconiosis, predominant reticular (n = 2), nodular (n = 2), and upper-lobe fibrosis (n = 2); and in hard-metal pneumoconiosis, multilobular ground-glass attenuation and consolidation with shrinkage (corresponding to marked intra-alveolar desquamation and multinucleated giant cells with mural mononuclear cell infiltrate). CONCLUSION: Predominant findings are characteristic in each type of pneumoconiosis and are depicted at thin-section CT. PMID- 7480685 TI - Characterization of adrenal masses with chemical shift and gadolinium-enhanced MR imaging. AB - PURPOSE: To assess the potential role of chemical shift and dynamic gadolinium enhanced magnetic resonance (MR) imaging in the characterization of adrenal masses. MATERIALS AND METHODS: Fifty-one adrenal masses (35 adenomas and 16 nonadenomas) in 43 patients were evaluated with chemical shift MR imaging, dynamic gadolinium-enhanced MR imaging, or both. The relative change in the signal intensity (SI) ratio of the adrenal mass to liver and paraspinal muscles was quantitatively and qualitatively assessed. Opposed-phase gradient-echo (GRE) images were compared with in-phase images. RESULTS: With qualitative visual inspection, only adenomas showed a decrease in relative SI ratio on opposed-phase chemical shift images (specificity, 100%; sensitivity, 81%). Quantitative ratios corresponding to 100% specificity were also observed, with similar sensitivities. Adenomas could not be differentiated from nonadenomas with visual assessment of maximum SI after contrast material administration or washout. CONCLUSION: Characterization of an adrenal mass as an adenoma can be made with high specificity and acceptable sensitivity by visually comparing opposed-phase with in-phase GRE images. PMID- 7480686 TI - Adrenal masses in patients with malignancy: prospective comparison of echo planar, fast spin-echo, and chemical shift MR imaging. AB - PURPOSE: To assess echo-planar, fast spin-echo (SE), and chemical shift magnetic resonance (MR) imaging in differentiation of adrenal adenomas from malignant adrenal masses in patients with known malignancy. MATERIALS AND METHODS: Sixty eight adrenal masses (23 malignant, 45 benign) in 68 patients with known malignancy were examined with echo-planar, fast SE with and without fat suppression, and chemical shift pulse sequences. RESULTS: With a cutoff T2 value of 75 msec, the sensitivity of echo-planar imaging for benign lesions was 82%, and specificity was 96%. With a cutoff adrenal mass-to-spleen signal intensity ratio of 0.80, the sensitivity of fast SE imaging for benign lesions was 53%, and specificity was 96%. With a cutoff adrenal mass-to-spleen ratio of 0.55, the sensitivity of chemical shift imaging for benign lesions was 80], and specificity was 100%. CONCLUSION: Chemical shift imaging and calculated T2 values from echo planar imaging are promising techniques for differentiation of adrenal adenomas from malignant adrenal masses and can obviate biopsy. PMID- 7480687 TI - Quantitative Doppler assessment of acute scrotal inflammation. AB - PURPOSE: To establish quantitative Doppler criteria for acute unilateral epididymitis and/or orchitis. MATERIALS AND METHODS: Triplex ultrasonography (US) was used to prospectively evaluate 31 patients with acute hemiscrotal pain and 15 asymptomatic control subjects. Morphology and perfusion were assessed, and peak systolic velocities (PSVs) were recorded. Ratios of PSV were calculated by using values from the right and left sides. RESULTS: PSVs in control subjects differed significantly from those in patients (P < .0001). A PSV > or = 15 cm/sec produced diagnostic accuracy of 90% for orchitis and 93% for epididymitis: Five of six false-negative findings were in patients younger than age 15 years. Epididymal PSV ratios > or = 1.7 or testicular ratios > or = 1.9 were diagnostic of acute inflammation (P < .0001). In three cases, morphologic and color Doppler findings were normal and only the PSV ratios were diagnostic. CONCLUSION: PSV and PSV ratios provide diagnostic criteria for acute unilateral epididymitis and/or orchitis that are more accurate than morphologic evaluation and color flow imaging either alone or in combination. PMID- 7480689 TI - Insufficiency fracture of the talus: diagnosis with MR imaging. AB - PURPOSE: To describe the clinical setting and pattern of talar insufficiency stress fractures as diagnosed with magnetic resonance (MR) imaging. MATERIALS AND METHODS: The authors retrospectively reviewed the clinical history and MR images from four patients with five talar insufficiency fractures. All patients were women (age range, 30-70 years). Images were evaluated for the location and orientation of fractures and the presence of concomitant osseous and soft-tissue injury. RESULTS: Concomitant fractures were identified in two patients. Only two fractures were typical in location and orientation at the talar neck. Three fractures were atypical in location and orientation. One occurred horizontally in the talar body and two occurred in the posteromedial talus (one was oriented vertically and one horizontally). Two patients had associated concomitant stress fractures--one in the distal tibia and one in the calcaneus. CONCLUSION: The location and orientation of talar insufficiency fractures is more variable than previously recognized. Specific diagnoses and coexisting fractures can be determined with MR imaging. PMID- 7480690 TI - US depiction of partial-thickness tear of the rotator cuff. AB - PURPOSE: To test previously defined ultrasound (US) criteria for identification of partial-thickness tears of the rotator cuff. MATERIALS AND METHODS: Before shoulder arthroscopy, 52 patients with shoulder pain for more than 3 months were examined with a 7.5-MHz commercially available linear-array transducer and a standardized study protocol. The criteria used to detect partial-thickness tears were (a) a mixed hyper- and hypoechoic focus in the crucial zone of the supraspinatus tendon and (b) a hypoechoic lesion visualized in two orthogonal imaging planes with either articular or bursal extension. RESULTS: The US findings were reported as partial-thickness tears in 17 shoulders, of which three were false-positive findings. There was one false-negative finding. The sensitivity of US in depiction of partial-thickness tears was 93%, and specificity was 94%. The positive predictive value was 82%, and the negative predictive value was 98%. CONCLUSION: US can depict most partial-thickness tears with use of the criteria described. PMID- 7480688 TI - Femoral head perfusion and composition: MR imaging and spectroscopic evaluation of patients with systemic lupus erythematosus and at risk for avascular necrosis. AB - PURPOSE: To measure fat content and relative marrow perfusion of the femoral head in patients at risk for avascular necrosis. MATERIALS AND METHODS: Thirteen patients with systemic lupus erythematosus (SLE) without avascular necrosis and with long-term corticosteroid treatment and 12 healthy subjects were evaluated. Hip perfusion was assessed with gadolinium-enhanced T1-weighted magnetic resonance (MR) imaging. Marrow composition in the femoral head and neck was measured with hydrogen-1 MR spectroscopy. RESULTS: Percentage fat content in the femoral head was similar between the two groups (SLE patients, 77% +/- 5 [standard deviation]; healthy subjects, 78% +/- 6; P = .74). Rate of enhancement of the femoral head was significantly higher in the SLE than the healthy group (11%/min +/- 6 vs 6.9%/min +/- 3, respectively; P = .05), was inversely related to fat content in healthy subjects (r = -.74, P = .02), and was more variable in the SLE group (r = -.53, P = .11). CONCLUSION: Perfusion of the femoral head is inversely related to marrow fat content in healthy and is higher in SLE patients without hip avascular necrosis. PMID- 7480691 TI - Cortical defect of the distal fibula: variant of ossification. AB - PURPOSE: To elucidate the nature of a normally occurring cortical defect of the distal fibula. MATERIALS AND METHODS: In patients with sprain and nonspecific ankle pain during a 2-year period, the authors reviewed ankle radiographs, computed tomographic (CT) scans, and magnetic resonance (MR) images. Cases were added from our file. Dried skeletons and cadavers were evaluated with radiography for the presence of this cortical defect. RESULTS: Fifteen (1.8%) of 847 radiographs in 739 patients (male, 377; female, 362; age range, 15-84 years) showed a small cortical defect in the anterior aspect at the level of the distal tibiofibular joint. The age range of the subjects with the defect (male, four; female, 11) was 16-66 years. MR imaging in two patients showed a small low-signal intensity area at the insertion site of the anterior tibiofibular ligament. Evaluation of 148 skeletons (75 subjects) revealed similar cortical defect in two cases (1.4%). CONCLUSION: The cortical defect of the distal fibula, at the insertion site of the anterior tibiofibular ligament, has no clinical significance and should not be confused with neoplasm. PMID- 7480692 TI - Osteoid osteoma: percutaneous radio-frequency ablation. AB - PURPOSE: To evaluate a percutaneous technique for in situ destruction of osteoid osteoma. MATERIALS AND METHODS: Radio-frequency ablation was performed in 18 patients with osteoid osteoma (17 male, one female; age range, 8-42 years). Diagnosis was established by means of clinical and radiographic features and confirmed by means of needle biopsy findings. No attempt was made to remove the lesion. A small radio-frequency electrode introduced into the lesion through the biopsy track was used to produce thermal necrosis of a 1-cm sphere of tissue. RESULTS: Symptoms were completely relieved in 16 (89%) of 18 patients. In one patient, a second procedure was required for pain relief. All but two patients underwent treatment as outpatients: These two were hospitalized for 1 night only. All patients resumed normal daily activities immediately. No casts or external supports were required; there were no complications. Twelve patients were followed up for more than 1 year. There were no recurrences CONCLUSION: Radio frequency ablation of osteoid osteoma is a promising alternative to surgery in selected patients. PMID- 7480693 TI - Iliac artery patency before and immediately after percutaneous transluminal angioplasty: assessment with time-of-flight MR angiography. AB - PURPOSE: To assess the efficacy of magnetic resonance (MR) angiography of iliac arteries before and immediately after percutaneous transluminal angioplasty (PTA). MATERIALS AND METHODS: In 14 patients with 22 diseased iliac artery segments (external or common), axial two-dimensional time-of-flight MR angiography was performed. Images were reconstructed with a maximum-intensity projection (MIP) algorithm. MR angiography was performed 1-4 days after diagnostic digital angiography and 6-24 hours after PTA. Findings obtained before and immediately after PTA were compared for number and location of significant (ie, > 50%) stenoses, length and diameter of balloon to be employed, and diameter of the stenotic artery after PTA. Linear regression analysis was performed. RESULTS: Sensitivity and specificity of MR angiography for determination of significant stenoses were 95% and 97%, respectively. Before PTA, balloon dimensions depicted on MR angiograms and digital angiograms were well correlated (r = .76, P < .05). After PTA, MR angiograms and digital angiograms provided similar findings in all but one case. CONCLUSION: MR angiography helped determine if PTA is indicated and depicted iliac artery patency after PTA. PMID- 7480694 TI - Study of thrombus from thrombosed hemodialysis access grafts. AB - PURPOSE: To determine the volume and composition of clot within thrombosed hemodialysis access grafts. MATERIALS AND METHODS: Clots were collected in 22 patients at surgical thrombectomy of polytetrafluoroethylene grafts. Histologic analysis was performed in 10 of these clots plus 21 randomly selected clots from the pathology archives. RESULTS: A small, firm piece of whitish thrombus ("arterial plug") was almost always recovered from the arterial limb of the graft. This plug had a concave surface and ranged from 5 mm to 3 cm in length. The remaining clot was soft, red thrombus. The mean weight of all clots was 3.4 g, and mean volume was 3.2 cm3. Average graft length was 42 cm. Histologically, the arterial plug had a characteristic appearance of densely compacted alternating layers of erythrocytes and fibrin. CONCLUSION: Clot volume in thrombosed dialysis grafts is much less (approximately equal to 25%) than would be expected if the graft were completely filled with thrombus, a finding of significance to mechanical thrombolytic techniques. Resistance of the arterial plug to pulse-spray thrombolysis is likely due to compaction. PMID- 7480695 TI - Renal vasculature in potential renal transplant donors: comparison of MR imaging and digital subtraction angiography. AB - PURPOSE: To compare magnetic resonance (MR) angiography with a selective inversion-recovery sequence and digital subtraction angiography (DSA) in renal transplant donors. MATERIALS AND METHODS: Thirteen potential donors underwent MR imaging at 1.5 T and conventional angiography. Blinded evaluation of imaging findings was performed. A selective inversion-recovery sequence was used to obtain MR arteriograms. RESULTS: Eight accessory vessels were present; MR angiography showed 100% sensitivity in accessory vessel detection. Both studies depicted early arterial branching in two vessels, mild fibromuscular dysplasia in one patient, normal renal size in all patients, and prominent fetal lobulation in two kidneys. A small cyst was detected with MR imaging only. Minor venous anomalies were noted in three patients, though these were not surgically confirmed owing to contralateral surgery. CONCLUSION: Despite the small study population, MR angiography with this sequence appears to have potential value in pretransplantation evaluation of the kidney. Advantages include short examination time, noninvasiveness, avoidance of iodinated contrast media, no radiation, and lower cost than DSA. PMID- 7480696 TI - Effectiveness of lead aprons in vascular radiology: results of clinical measurements. AB - PURPOSE: To determine the effectiveness of 0.5-mm-thick lead-equivalent aprons in vascular radiology. MATERIALS AND METHODS: The radiation dose to radiology personnel was measured under and over lead aprons in more than 100 patient examinations. Thermoluminescent dosimeters of high and normal sensitivities (detection levels of 1 and 10 microGy, respectively) were used. Dose measurements in clinical practice were complemented with real-time measurement of generating potential, dose-area product, and fluoroscopy time. In addition, measurements of the radiation attenuation of aprons were performed in simulated exposure conditions. The generating potential in fluoroscopy--weighted for the contribution to the occupational exposure--varied between about 70 and 95 kV per examination (average, 78 kV). RESULTS: On the basis of measurements obtained under clinical conditions in vascular radiology, attenuation factors between 70 and 280 were derived. The typical radiation attenuation factor for 0.5-mm-thick lead-equivalent aprons was 200. CONCLUSION: The effectiveness of 0.5-mm-thick lead-equivalent aprons, determined with clinical measurements, agrees with estimates based on generating potential. Previously published effective doses, inferred from nonclinically determined radiation transmission factors, probably overestimate the actual dose levels. PMID- 7480698 TI - Real-time biplanar needle tracking for interventional MR imaging procedures. AB - PURPOSE: To evaluate real-time biplanar tracking of a specially designed needle with magnetic resonance (MR) imaging guidance. MATERIALS AND METHODS: The needle is made of polyetheretherketone and has a miniature radio-frequency coil incorporated into the tip. Tracking software on two workstations is used to compute three-dimensional coordinates of the coil and to display the position as a moving symbol in two imaging planes. Validation of needle tracking was performed in a harvested human liver. T2-weighted fast spin-echo images were used to target a 1-cm cyst. Success of needle placement was confirmed with aspiration and with updated gradient-recalled-echo images. RESULTS: The cyst was successfully targeted from different approaches. Tracking procedures were monitored in real time simultaneously on two separate images. CONCLUSION: Real time biplanar needle tracking may prove to be useful for both diagnostic and therapeutic interventional MR imaging procedures. PMID- 7480697 TI - Late reobstruction in iliac arterial stents: percutaneous treatment. AB - PURPOSE: To retrospectively analyze in a nonrandomized fashion the efficacy of percutaneous reintervention in obstructed iliac stents. MATERIALS AND METHODS: In 21 symptomatic patients with iliac lesions, 26 reinterventions (16 for stent occlusion and 10 for stent stenosis) were performed. Restenoses were treated with balloon dilation and either atherectomy or stent placement. Reocclusions were treated with atherectomy or aspiration thrombectomy and then recanalization with balloon dilation and selective stent placement. RESULTS: Balloon angioplasty for stent stenosis was effective in all but one patient. Recanalization was successful in 14 of 16 patients with stent occlusion. The mean period of patency after reintervention was 18 months +/- 15. Cumulative stent stenosis patency after reintervention was 87% after 1 year. Stent occlusion patency was 57%. Recurrent stent obstruction occurred in eight of 24 (33%) patients with successful primary interventions. CONCLUSION: Percutaneous reintervention for both stent stenosis and occlusion is feasible with a moderate complication rate and may be attempted before surgery. PMID- 7480699 TI - Fetal liver calcifications: sonographic appearance and postnatal outcome. AB - PURPOSE: To determine the importance of intrahepatic calcifications seen sonographically in the fetus. MATERIALS AND METHODS: The authors retrospectively reviewed sonograms obtained in fetuses scanned between 16 and 40 gestational weeks in whom intrahepatic calcifications were seen. The number and character of these calcifications were determined. Follow-up scans obtained in utero and after birth were also reviewed. Outcome was determined after birth. RESULTS: Thirty three fetuses had intrahepatic calcifications at 16-38 weeks gestation. Four of the 33 fetuses died, one of which had liver calcifications as the only sonographic finding. This fetus had cytomegalovirus and was the only fetus with increasing numbers of calcifications on follow-up scans. All fetuses with isolated liver calcifications had a normal postnatal outcome (96% survival rate for fetuses with isolated intrahepatic calcifications). Of eight fetuses with additional anomalies, five lived; only two were healthy newborns. CONCLUSION: The outcome in fetuses with isolated intrahepatic calcifications is usually excellent, although viral causes must be excluded if additional findings appear. PMID- 7480700 TI - Intussusception: ability of fluoroscopic images obtained during air enemas to depict lead points and other abnormalities. AB - PURPOSE: To evaluate the ability of fluoroscopic images obtained during air enemas to depict or exclude lead points of intussusceptions and other abnormalities that require surgical treatment in the absence of intussusception. MATERIALS AND METHODS: The clinical, radiologic, surgical, and pathologic findings were reviewed in 14 patients with lead points or other lesions. RESULTS: Fluoroscopic images failed to depict a lead point in 10 patients. The air enema easily reduced intussusceptions with benign lead points in seven patients. Fluoroscopic images depicted pathologic lead points in two patients and were normal in two patients with intussusception. Fluoroscopic images failed to depict an abnormality that required surgery in the absence of intussusception in two patients. CONCLUSION: Successful reduction of an intussusception does not always rule out a lead point. Other imaging studies, the patient's condition, and laparotomy may also be necessary to diagnose and treat lead points and other lesions. PMID- 7480701 TI - Frontal foramina in pediatric skull in cases of congenital hydrocephalus. AB - PURPOSE: To describe a new observation, frontal calvarial foramina, in pediatric patients with congenital hydrocephalus secondary to central nervous system malformation. MATERIALS AND METHODS: Frontal foramina were initially identified in three female patients with Chiari II malformation. Subsequently, head computed tomographic (CT) scans in 99 patients with congenital hydrocephalus were retrospectively reviewed. CT scans in a control group of 116 patients without hydrocephalus were also retrospectively reviewed. RESULTS: Frontal foramina were found in eight of 61 (13%) patients with Chiari II malformation, in one child with Dandy-Walker malformation, and in one child with occipital horn dilatation (colpocephaly), but not in control patients. Sequential CT examinations in three patients with frontal foramina depicted gradual closure after ventriculoperitoneal shunt placement. CONCLUSION: Frontal foramina may represent an abnormality variably expressed in certain central nervous system malformations that cause congenital hydrocephalus. The presence of frontal foramina palpated or visualized on plain radiographs may help in the diagnosis of congenital hydrocephalus and central nervous system malformation. PMID- 7480702 TI - Soft-copy computed radiography in neonatal and pediatric intensive care units: cost-savings analysis. AB - PURPOSE: To determine whether soft-copy interpretation of computed radiography (CR) in neonatal and pediatric intensive care units (ICUs) can result in a cost savings compared with screen-film radiography for the radiology department. MATERIALS AND METHODS: The actual cost of CR equipment, maintenance contracts, and the estimated cost for data storage were compared with the potential savings that could result from eliminating film, decreasing the number of librarians and lost radiographs, and improving technologists' efficiency. RESULTS: After the 1st year, net cash flow from use of soft-copy CR interpretation is projected to result in a slight savings for the radiology department but not enough to offset the capital equipment purchase price. CONCLUSION: Soft-copy CR ICU imaging will not result in a cost savings. To have a balanced net cash flow would require either more than doubling potential savings or decreasing capital equipment cost by greater than one-half. The justification for soft-copy CR needs to be judged by its effect on increasing physician efficiency and timely decisions on patient care. PMID- 7480703 TI - Omission of axillary lymph node dissection in early-stage breast cancer: effect on treatment outcome. AB - PURPOSE: To determine the effect omission of axillary lymph node dissection has on outcome in patients treated with breast-conserving therapy for early-stage invasive breast cancer. MATERIALS AND METHODS: The authors evaluated 492 patients with breast cancer treated with (n = 32) and without (n = 456) axillary lymph node dissection. The primary tumor characteristics of the two groups were similar, though the median age was different. All patients received whole-breast radiation (mean dose, 50 Gy); additional tumor bed boosts and nodal irradiation were used more often in patients without dissection. RESULTS: Median follow-up in patients without and with dissection was 60 and 52 months, respectively. The 5 year survival was 88% and 93%, respectively. There were no regional failures in the group treated without dissection. Crude rates of local and distant failure were similar for both groups. CONCLUSION: Omission of axillary lymph node dissection should be considered in patients whose pathologic nodal status will not influence decisions regarding adjuvant therapy. PMID- 7480704 TI - Cholangiocarcinoma: clinical significance of tumor location along the extrahepatic bile duct. AB - PURPOSE: To define the criteria for resection and/or radiation therapy (RT) of extrahepatic bile duct cancer. MATERIALS AND METHODS: Of 81 patients with extrahepatic bile duct cancer treated from 1983 to 1992, those with proximal duct lesions (n = 56) underwent RT and/or resection or palliative care, and those with distal lesions (n = 25) underwent resection with or without RT. Follow-up was available 3-114 months (median, 28 months). RESULTS: Patients with distal bile duct cancer lived longer than patients with proximal bile duct cancer (survival with Kaplan-Meier analysis, 53% vs 13% at 5 years, respectively, P < .01). Median survival in patients with proximal cancer after RT was more than double that without RT (17 months vs 6 months, respectively, regardless of stage [P = .01]); survival was not significantly different after resection. In patients with distal cancer, RT after resection made no significant difference in median survival (68 months). CONCLUSION: Patients with proximal cancer should undergo primary RT, and expectations should be limited. Patients with distal cancer should undergo resection, and RT may not be needed. PMID- 7480706 TI - Brain lesions in patients with AIDS: H-1 MR spectroscopy. AB - PURPOSE: To evaluate the role of proton (hydrogen-1) magnetic resonance (MR) spectroscopy in the differential diagnosis of focal brain lesions in patients with acquired immunodeficiency syndrome (AIDS). MATERIALS AND METHODS: Twenty-six men with 35 AIDS-related brain lesions underwent MR imaging and localized H-1 MR spectroscopy. Lesions consisted of 11 toxoplasmic abscesses, 12 progressive multifocal leukoencephalopathic lesions, eight lymphomas, and four cryptococcomas. Metabolite peak areas in the lesions were compared with those in the contralateral hemisphere in each patient. RESULTS: H-1 MR spectroscopic findings showed significantly different biochemical profiles for each diagnostic group (P = .0001) with regard to N-acetyl compounds, total creatine pool, choline containing compounds, myoinositol, and lactate. H-1 MR spectroscopy alone helped correctly diagnose 94% (84% with jackknifed classification) of the brain lesions, without overlap between toxoplasmosis and lymphoma. CONCLUSION: H-1 MR spectroscopy is a sensitive and potentially specific noninvasive adjunctive method for differential diagnosis of focal brain lesions in AIDS. PMID- 7480705 TI - H-1 MR spectroscopic imaging of white matter signal hyperintensities: Alzheimer disease and ischemic vascular dementia. AB - PURPOSE: To investigate the association of white matter signal hyperintensities (WMSHs) with changes in hydrogen-1 metabolites. MATERIALS AND METHODS: T2 weighted magnetic resonance (MR) imaging and H-1 MR spectroscopic imaging were performed in 21 elderly control subjects without or with minimal WMSHs, eight elderly subjects with substantial WMSHs, 11 probable Alzheimer disease patients with WMSHs, and eight ischemic vascular dementia (IVD) patients with WMSHs. N acetylaspartate (NAA), choline-containing metabolites (Cho), and creatine containing metabolites (Cr) were analyzed. RESULTS: Differences in regional metabolite levels were found within the supraventricular brain of elderly control subjects. In Alzheimer disease patients, extensive WMSHs showed a lower percentage of NAA and a higher percentage of Cho compared with contralateral normal-appearing white matter (NAWM); in IVD patients, extensive and large WMSHs were associated with a higher percentage of Cho and a lower percentage of Cr compared with contralateral NAWM. CONCLUSION: Regional metabolite variation and the presence of WMSHs are important covariants that must be accounted for in analysis of MR spectroscopic data. PMID- 7480707 TI - MR imaging of phagocytosis in experimental gliomas. AB - PURPOSE: To determine whether phagocytosis can be observed in vivo in glioma cells. MATERIALS AND METHODS: Rat C6 glioma cells were studied in culture and after intracerebral implantation into 13 rats. Monocrystalline iron oxide nanoparticles (MION), a model marker of phagocytosis, was administered intravenously to tumor-bearing rats at 2-20 mg of iron per kilogram. Magnetic resonance (MR) imaging was performed at multiple time points. RESULTS: Glioma cells in culture showed uptake of MION in amounts of up to 10 ng of iron per 10(6) cells, corresponding to approximately 50,000 particles per cell. Fluorescently labeled MION was found to be located primarily in tubular lysosomes. Intracerebral gliomas showed characteristic changes in signal intensity at MR imaging that peaked 12 hours after administration of MION and lasted up to 5 days; these changes corresponded to uptake and subsequent biodegradation of MION by tumor cells. CONCLUSION: Phagocytosis of glioma cells can be detected in vivo with iron oxide-enhanced MR imaging, and this may permit accurate delineation of tumor margins. PMID- 7480708 TI - Reproducibility of quantitative, spirometrically controlled CT. AB - Quantitative spirometrically controlled computed tomography (with 1-mm-thick sections) was performed twice (with a 5-minute break) in 24 adult patients with pulmonary disease to objectively evaluate parenchymal changes in the lung. Twelve measurements of attenuation were made on apical, carinal, and basal scans (right, left, total of each level, total right, total left, total of all three scans), obtained at 50% vital capacity. Since differences in measurements between the first and second examination were not significant, the method provides highly reproducible results. PMID- 7480710 TI - New contraindication to intravascular iodinated contrast material. PMID- 7480709 TI - Peripherally inserted central venous catheters: US-guided vascular access in pediatric patients. AB - Ultrasound (US)-guided peripheral venipuncture was performed for peripheral insertion of 222 central venous catheters over a 12-month period. Initial placement was successful in 218 patients but unsuccessful in eight; placement was successful in four the next day (success rate, 98%; complication rate, 5%). Catheters were in place from 3 days to 6 months (mean, 36 days). US guidance allowed successful venipuncture for placement of central venous catheters in children. PMID- 7480711 TI - New contraindication to intravascular iodinated contrast material. PMID- 7480712 TI - Intraosseous ganglia of the wrist. PMID- 7480713 TI - Nephrotoxicity from contrast media: attenuation with theophylline. PMID- 7480714 TI - Nephrotoxicity from contrast media: attenuation with theophylline. PMID- 7480715 TI - Malpractice lawsuits in radiology: comparison of the Italian and American situations. PMID- 7480716 TI - Recertification. PMID- 7480717 TI - Pulmonary thromboembolism: recent developments in diagnosis with CT and MR imaging. AB - Recent technical advances in computed tomography (CT; helical and electron beam) and magnetic resonance (MR) imaging have spurred a renewed interest in these modalities for the diagnosis of acute and chronic pulmonary embolism (PE). These techniques can enable accurate clot detection down to segmental pulmonary arteries, with CT currently allowing more accuracy than that with MR imaging. Ongoing technical advances, particularly in MR angiography, will likely increase diagnostic accuracy. Inability to reliably detect subsegmental acute emboli may not prove to be a clinically significant limitation if lung imaging is coupled with evaluation for deep venous thrombosis. MR imaging can potentially accomplish this within a single examination. Incorporation of CT and MR imaging into diagnostic algorithms for suspected PE can be cost-effective. Evaluation of these new modalities should be based on patient outcome, not solely on clot detectability. Well-designed clinical trials are warranted before CT and MR imaging can be used routinely in the diagnosis of acute PE. PMID- 7480718 TI - Technique for MR imaging of the liver. PMID- 7480719 TI - Thin-section three-dimensional spiral CT: is isotropic imaging possible? PMID- 7480720 TI - Role of duplex US as a screening test for carotid atherosclerotic disease: benefit without cost? PMID- 7480721 TI - Management of small renal neoplasms and angiomyolipoma: a growing problem. PMID- 7480722 TI - Risk of intentional reversible thrombosis of transjugular intrahepatic portosystemic shunt. PMID- 7480723 TI - Judicious use of intentional reversible thrombosis of transjugular intrahepatic portosystemic shunt and renal blood flow. PMID- 7480724 TI - Small renal parenchymal neoplasms: further observations on growth. AB - PURPOSE: To determine the growTH rate and behavior of small (< or = 3.5-cm diameter) incidentally detected renal parenchymal neoplasms. MATERIALS AND METHODS: Forty renal parenchymal tumors in 37 adult patients (mean age, 65.5 years) were observed with computed tomography (CT) and ultrasound for 1.75-8.5 years (mean, 3.25 years). Surgical removal was performed of 26 tumors proved and graded at pathologic examination. Fourteen tumors with CT criteria of neoplasm are being followed up. RESULTS: The overall growth rate of neoplasms was 0-1.1 cm/y (mean, 0.36 cm/y). Thirty tumors grew 0.5 cm/y or less and 19 grew very slowly (< or = 0.35 cm/y). No patient developed metastatic disease. Nine (24%) had multiple neoplasms. CONCLUSION: Most small, incidentally discovered, well marginated renal parenchymal neoplasms grow slowly and are not an immediate threat to a patient's life. Watchful waiting might be appropriate instead of surgical removal, especially in patients who are elderly or may not survive surgery. PMID- 7480725 TI - Renal angiomyolipoma: growth followed up with CT and/or US. AB - PURPOSE: To follow up the growth of renal angiomyolipomas (AMLs). MATERIALS AND METHODS: Patients with known AML (n = 55) were divided into three groups on the basis of initial clinical and computed tomographic (CT) findings: group 1, isolated AML (n = 43); group 2, multiple AMLs without tuberous sclerosis (TS) (n = 6); group 3, multiple AMLs with TS (n = 6). Follow-up ultrasonography (US) and CT were performed in 55 and 31 patients, respectively. Growth of the AMLs (n = 59) was evaluated on the basis of area on initial and follow-up images. RESULTS: Mean percentage growth was 17%, 128%, and 47%, and mean growth rate per year was 5%, 22%, and 18% in groups 1, 2, and 3, respectively. New renal lesions were noted in three patients in groups 1 and 2, but no new lesions were detected in group 3, because of the large number of AMLs. New extrarenal lesions were observed in four, two, and three patients in groups 1, 2, and 3, respectively. No correlation was found between percentage of fatty tissue and growth rate. CONCLUSION: Multiple AMLs show more growth than solitary AMLs. PMID- 7480726 TI - Change in endometrial thickness in postmenopausal women undergoing hormone replacement therapy. AB - PURPOSE: To evaluate the effect over time of hormone replacement on endometrial thickness in asymptomatic postmenopausal women. MATERIALS AND METHODS: Endometrial thickness was measured on 644 sonograms of 120 postmenopausal volunteers receiving no hormones or one of three hormone regimens. Each subject underwent ultrasound (US) two to nine times. In addition, eight women using sequential hormones underwent US seven to 20 times over 2 months. The greatest endometrial thickness and change in endometrial thickness over time were evaluated. RESULTS: Endometrial thickness was statistically significantly greater in women using sequential hormones (8 mm) compared with that in control subjects (5 mm). Change in endometrial thickness (> 3-mm difference) was seen in 23 of 40 women using sequential hormones compared with six of 61 controls. Women using sequential hormones demonstrated maximum endometrial thickness on days 13-23 of the cycle. CONCLUSION: Women using sequential hormones show greater endometrial thickness than that in controls and show the most variation in measurements. They should undergo US either early or late in the hormone cycle to evaluate the endometrium at its thinnest. PMID- 7480727 TI - Diffuse uterine adenomyosis: morphologic criteria and diagnostic accuracy of endovaginal sonography. AB - PURPOSE: To determine the accuracy of endovaginal ultrasound (US) in the diagnosis of uterine adenomyosis and to evaluate the frequency of observed sonographic criteria. MATERIALS AND METHODS: Endovaginal US was performed in 100 consecutive women undergoing hysterectomy for a variety of benign and malignant conditions. Adenomyosis was diagnosed when a poorly defined area of abnormal echotexture (decreased or increased echogenicity, heterogeneous echotexture, myometrial cysts) was present in the myometrium. All endovaginal US findings were correlated with those from histologic examination. RESULTS: Endovaginal US depicted 25 of 29 pathologically proved cases of adenomyosis. Adenomyosis was correctly ruled out in 61 of 71 patients. Endovaginal US had a sensitivity of 86%, a specificity of 86%, and a positive and negative predictive value of 71% and 94%, respectively. Of the 25 patients with true-positive findings at US, the myometrium demonstrated heterogeneous and hypoechoic areas with or without the presence of cysts in 21 (84%) patients, hypoechoic areas with cysts in three (12%) patients, and heterogeneous areas within the myometrium in one (4%) patient. CONCLUSION: Adenomyosis of the uterus can be accurately diagnosed with endovaginal US with use of specific sonographic criteria. PMID- 7480728 TI - Renal perfusion: pharmacologic changes depicted with power Doppler US in an animal model. AB - PURPOSE: To evaluate the ability of power Doppler ultrasonography to depict changes in renal perfusion after pharmacologic manipulations. MATERIALS AND METHODS: Six domestic swine were intubated and anesthetized. The right renal artery was catheterized for injection of epinephrine (100 micrograms) and papaverine (2 mg/kg). Intraarterial blood pressure, heart rate, and oxygen saturation were measured continuously. Longitudinal images of the kidney were obtained, and the power Doppler color map was analyzed subjectively and by computer at baseline and at the time of maximum drug effect. RESULTS: Epinephrine produced vasoconstriction with a diminished power Doppler color map and a decrease in both subjective and computer scores. After papaverine-induced vasodilatation, the power Doppler map was enhanced, as reflected by increased subjective and computer scores. CONCLUSION: The power Doppler color map reflects anticipated changes in renal perfusion after alterations in blood flow by vasoactive drugs. PMID- 7480729 TI - Ovarian cancer: staging with CT and MR imaging. AB - PURPOSE: To evaluate ovarian cancer staging and tumor resectability with computed tomography (CT) or magnetic resonance (MR) imaging. MATERIALS AND METHODS: Eighty two women underwent CT (n = 43) or MR imaging (n = 50); eleven of these 82 underwent both. Imaging was performed within 4 weeks of surgical staging. Radiologic, surgical, and histopathologic findings were compared. RESULTS: Overall staging accuracy was similar for CT and MR imaging (77% [33 of 43] vs 78% [39 of 50]). Evaluation of pelvic cancer extent was better with MR imaging than with CT. There was no difference in detection of abdominal disease. Most mesenteric and small-bowel implants were not detected with either CT or MR imaging. For CT, the positive predictive value for cancer nonresectability was 100% (three of three patients); the negative predictive value was 92% (37 of 40 patients). The positive and negative predictive values for MR imaging were 91% (10 of 11 patients) and 97% (38 of 39 patients). CONCLUSION: While the staging accuracy of both CT and MR imaging is only moderate, prediction of tumor resectability is excellent. PMID- 7480730 TI - Hyperparathyroidism in high-risk surgical patients: evaluation with double-phase technetium-99m sestamibi imaging. AB - PURPOSE: To evaluate the accuracy of technetium-99m sestamibi as a single agent in the detection and localization of hyperfunctioning parathyroid tissue in patients who underwent prior neck exploration or who otherwise are high surgical risks. MATERIALS AND METHODS: Thirty-nine patients with hyperparathyroidism underwent 40 double-phase Tc-99m sestamibi studies. Histopathologic correlation was obtained for all studies. RESULTS: Thirty patients had solitary adenomas, and nine had hyperplastic glands. One developed recurrent hypercalcemia after five gland resection and underwent repeat imaging and operation. Double-phase sestamibi imaging prospectively enabled localization of 46 (77%) of 60 abnormal glands. Twenty-eight (93%) of 30 adenomas were correctly localized, whereas 18 (60%) of 30 hyperplastic glands were localized. The overall specificity of the study was 98% with one false-positive study. Gland weight and vascularity were statistically significant predictors of uptake of sestamibi. CONCLUSION: Double phase Tc-99m sestamibi imaging is a promising technique for localization of parathyroid adenomas in high-risk surgical patients. Localization of multiple hyperplastic glands remains a challenge. PMID- 7480731 TI - Role of Doppler US in screening for carotid atherosclerotic disease. AB - PURPOSE: To evaluate Doppler ultrasound (US) as a screening modality before arteriography for extracranial carotid artery disease. MATERIALS AND METHODS: The net benefit in stroke reduction from screening with Doppler US was calculated on the basis of literature estimates of disease prevalence, risk reduction data, and locally validated sensitivities and specificities for detection of carotid artery stenosis and occlusion in 215 patients. RESULTS: Screening a symptomatic population demonstrated a net stroke reduction. Screening asymptomatic populations with a 20% prevalence of > or = 60% stenosis also yielded a net stroke reduction. Screening low-prevalence (5%) asymptomatic populations produced a small benefit, which was lost if arteriographic or surgical complications increased slightly. Arteriographic confirmation of the US diagnosis of occlusion produced a small benefit only in the symptomatic population. CONCLUSION: Screening symptomatic and high-prevalence asymptomatic populations with US reduces stroke. Increased arteriographic or surgical complication rates reduce the benefit of screening in any population. PMID- 7480732 TI - Three-dimensional craniocervical helical CT: is isotropic imaging possible? AB - PURPOSE: To assess whether isotropic resolution is attainable in three dimensional craniocervical helical computed tomography (CT). MATERIALS AND METHODS: Two cadaveric cervical spine and two orbital phantoms were scanned in air and water by using 180 degrees-interpolated helical CT with three-dimensional reconstruction. Scan parameters yielded nominally isotropic voxels with reconstruction of 1-mm collimated and incremental sections at 0.2-mm intervals. CT scan angle was varied with the assumption that volume-averaging artifacts should be scan angle-independent with isotropic voxels. Twelve blinded observers analyzed three-dimensional images generated from multiple CT orientations to assess changes in volume-averaging artifacts that would indicate voxel anisotropy. RESULTS: Significant observable differences in three-dimensional image representation were present in test anatomic regions in one cervical spine phantom (two-tailed P < .006) and in both orbital phantoms (P < .008 and P < .006). No statistically significant differences were present for control anatomic regions. CONCLUSION: Isotropic scanning is not possible in three-dimensional craniocervical helical CT. PMID- 7480733 TI - Toxoplasmosis and primary central nervous system lymphoma in HIV infection: diagnosis with MR spectroscopy. AB - PURPOSE: To differentiate intracranial lymphoma from Toxoplasma gondii lesions in patients infected with the human immunodeficiency virus, by means of localized spin-echo proton magnetic resonance (MR) spectroscopy. MATERIALS AND METHODS: Twenty-seven lesions were studied (18 T gondii lesions, nine lymphoma lesions) at 1.5 T. Spectra were acquired at an echo time of 135 msec from voxels centered on the lesions. Both visual analysis and spectral fitting were used to obtain metabolite ratios for choline (Cho), creatine (Cr), N-acetyl (NA), lactate, and lipids. RESULTS: Three spectral categories were seen. One had large lipid peaks with suppression of other metabolites. Another had an elevated Cho/Cr ratio with relatively diminished NA. The third had features of the other two. Examples of each spectrum type were acquired from both T gondii and lymphoma lesions. Neither method of analysis allowed differentiation between lesion types. MR spectroscopy showed an overlap of spectra. CONCLUSION: The authors conclude that toxoplasmosis and lymphoma cannot be differentiated with spin-echo proton MR spectroscopy at 135 msec. PMID- 7480734 TI - Spinal cord ependymomas: MR imaging features. AB - PURPOSE: To assess the magnetic resonance (MR) imaging characteristics of spinal intramedullary ependymomas. MATERIALS AND METHODS: MR images obtained in 25 patients (aged 12-73 years) with proved intramedullary ependymomas were retrospectively reviewed. T1- and T2-weighted images were obtained in all patients. Gadopentetate dimeglumine was intravenously administered in 23 patients; enhanced sagittal and axial T1-weighted spin-echo images were reviewed. RESULTS: All tumors had hyperintense signal on T2-weighted images. In the 23 patients who received contrast material, all tumors became enhanced; enhancement was heterogeneous in 15 patients and homogeneous in eight patients. Twenty tumors had sharply defined, enhanced borders. Nineteen tumors were centrally located in the spinal cord. A hypointense rim on T2-weighted images was noted in five patients. CONCLUSION: Intramedullary ependymomas become enhanced after administration of gadopentetate dimeglumine; the enhanced borders are usually sharply marginated. They are characteristically located centrally in an expanded spinal cord. Hemosiderin is often present at the periphery of cervical ependymomas. PMID- 7480735 TI - Hemodynamic alterations in the paravertebral venous plexus after spinal injury. AB - PURPOSE: To assess the prevalence of altered venous hemodynamics after spinal cord injury. MATERIALS AND METHODS: The authors performed venography in 200 ambulant patients and 107 patients with spinal injury. The hemodynamic characteristics of the paravertebral venous plexus were documented after pedal injection of contrast material. The hemodynamic and clinical data were correlated in patients with spinal cord injury. RESULTS: Abnormal venous return via the paravertebral venous plexus in the absence of iliac or inferior vena caval occlusion was shown in 73 (68%) of the 107 patients with spinal injury. This paravertebral venous flow route is not related to the time between injury and examination. It was demonstrable in the early as well as late post-traumatic period and was reproducible at subsequent examinations. The paravertebral venous flow route was more commonly seen in cervical than thoracolumbar injuries and was closely related to the severity of neurologic deficit (P = .0012). CONCLUSION: Abnormal vertebral venous hemodynamics occur in most patients with spinal cord injury and are more likely to occur in patients with complete neurologic deficit and cervical injury. PMID- 7480737 TI - Anal sphincter complex: endoanal MR imaging of normal anatomy. AB - PURPOSE: To determine the normal anatomy of the anal sphincter complex on magnetic resonance (MR) images. MATERIALS AND METHODS: Ten healthy volunteers (four men, six women; age range, 21-26 years) underwent MR imaging with an endoanal coil. RESULTS: The lower part of the anal canal contained the internal sphincter, the longitudinal muscle layer, and the external sphincter; the upper part comprised the internal sphincter, the longitudinal layer, and the puborectal muscle. At the upper end, the puborectal muscle was attached to the levator ani muscle. Anteriorly, the external sphincter was connected to the urogenital diaphragm; posteriorly, it was attached to the coccyx with the anococcygeal ligament. All perianal spaces were visible. The morphology of the anterior part of the external sphincter, different in men and women, was well displayed. CONCLUSION: Depiction of the anal sphincter complex on MR images is excellent. PMID- 7480736 TI - Inflammation: imaging with methoxy poly(ethylene glycol)-poly-L-lysine-DTPA, a long-circulating graft copolymer. AB - PURPOSE: To test whether a nontargeted, long-circulating, synthetic polymer accumulates in areas of inflammation, with high capillary permeability and increased regional blood flow. MATERIALS AND METHODS: Methoxy poly(ethylene glycol)-poly-L-lysine (PL)-diethylenetriaminepentaacetic acid (MPEG-PL-DTPA) was labeled with technetium-99m for scintigraphy and with gadolinium for magnetic resonance (MR) imaging. Eleven Escherichia coli-infected rats were injected with 1.0 mCi (37 MBq) of Tc-99m-labeled MPEG-PL-DTPA for scintigraphy. Twelve rats underwent 1.5-T MR imaging after intravenous injection of gadolinium-labeled MPEG PL-DTPA (35 mumol/kg). RESULTS: Tc-99m-labeled MPEG-PL-DTPA demonstrated nearly eight-fold higher accumulation in infected muscle when compared with normal muscle. Scintigrams and MR images showed areas of inflammation with peak accumulation at 24 hours after injection of Tc-99m- or gadolinium-labeled MPEG-PL DTPA. CONCLUSION: Nontargeted, long-circulating, copolymers can efficiently accumulate in sites of inflammation and thus represent an alternative to inflammation-specific agents. PMID- 7480738 TI - Proctographic features of anismus. AB - PURPOSE: To document the proctographic features of anismus at evacuation proctography and determine the optimum radiologic measurements for diagnosis. MATERIALS AND METHODS: Twenty-four patients with anismus according to clinical and multiple physiologic criteria were examined with evacuation proctography. Structural and functional measurements were compared with those of a group of 20 asymptomatic subjects. RESULTS: No significant difference between patients and control subjects was found with respect to pelvic descent, rectocele, or any anorectal angle measurement. In patients with anismus, initiation of evacuation was prolonged (median, 9 vs 3 seconds for control subjects; P < .0001) and anal canal width was reduced (median, 0.6 vs 1.2 cm; P = .0075). Evacuation time was increased (median, 50 vs 10 seconds; P < .0001), and the percentage of contrast material evacuated was decreased (median, 60% vs 100%; P < .0001). Only four patients were able to evacuate more than 66% of the contrast material within 30 seconds, whereas all control subjects were able to do so. CONCLUSION: Measurement of the anorectal angle to diagnose anismus should be abandoned. Patients with anismus demonstrate delayed initiation of evacuation, which is also prolonged and incomplete. Incomplete evacuation after 30 seconds is highly suggestive of anismus. PMID- 7480739 TI - Colon cancer: morphology detected with barium enema examination versus histopathologic stage. AB - PURPOSE: To determine the relationship between the morphology of colon carcinomas detected with barium enema examination and the cancer stage. MATERIALS AND METHODS: Clinical, radiographic, endoscopic, surgical, and histopathologic findings were retrospectively reviewed in 152 patients with colon cancer detected with barium enema examination during a 2-year period. RESULTS: Eighty-six patients (57%) had lesions in the rectum and sigmoid and descending colon, and 66 (43%) patients had lesions more proximally in the colon. Lesions on the right side of the colon were less likely to cause symptoms than those on the left side. Eighty-one patients (53%) had annular or semiannular lesions, 57 (38%) had polypoid lesions, and 14 (9%) had plaquelike or carpet lesions. Six patients (4%) had Dukes stage A lesions; 84 (55%), Dukes stage B lesions; 42 (28%), Dukes stage C lesions; and 20 (13%), Dukes stage D lesions. CONCLUSION: Annular or semiannular carcinomas had higher rates of serosal invasion and lymph node metastasis than polypoid carcinomas, but the rates of liver metastases were comparable. PMID- 7480740 TI - Detection of pelvic recurrence of colorectal carcinoma: prospective, blinded comparison of Tc-99m-IMMU-4 monoclonal antibody scanning and CT. AB - PURPOSE: To prospectively compare the accuracy of imaging with technetium-99m labeled Fab' fragment of the anti-carcinoembryonic antigen antibody (CEA) IMMU-4 with that of computed tomography (CT) for the detection of pelvic recurrence of colorectal carcinoma. MATERIALS AND METHODS: In 61 patients, blinded interpretations of both modalities were correlated with surgical-pathologic (n = 23) or clinical and CT follow-up findings (n = 38). RESULTS: Sensitivity and specificity with antibody scanning alone and combined with CT (79% and 84% vs 83% and 81%, respectively) were not significantly different from those values for CT alone (66% and 97%, respectively). Sensitivity of antibody scanning was greater for recurrences larger than 2 cm (94% vs 55% [P = .02]), serum CEA more than 2.5 ng/mL (91% vs 40% [P = .03]), and combined planar and single photon emission CT antibody scanning compared with planar alone (79% vs 48% [P = .03]), without a significant decrease in specificity. CONCLUSION: Antibody scanning does not improve on findings at CT alone for recurrent colorectal carcinoma but can help differentiate recurrent tumor from fibrosis. PMID- 7480741 TI - Pathologic primer on gastritis: an illustrated sum and substance. PMID- 7480742 TI - Radiologic, endoscopic, and surgical gastrostomy: an institutional evaluation and meta-analysis of the literature. AB - PURPOSE: To evaluate the effectiveness and safety of radiologic, percutaneous endoscopic (PEG), and surgical gastrostomy. MATERIALS AND METHODS: This project involved 5,752 patients (837 patients underwent radiologic gastrotomy; 4,194, PEG; and 721, surgical gastrostomy). Seventy-two (47 male, 25 female; age range, 12-94 years) underwent gastrostomy within 1 year in this series (radiologic gastrostomy, n = 33; PEG, n = 35; surgical gastrostomy, n = 4). A meta-analysis of 5,680 additional cases from literature published from 1980 to the present was also performed. RESULTS: Rates of successful tube placement were higher for radiologic gastrostomy than for PEG in our series and in the meta-analysis (99.2% vs 95.7%, P < .001). Major complications occurred less frequently after radiologic gastrostomy in our series and in the meta-analysis (5.9% vs 9.4% for PEG and 19.9% for surgery, P < .001). Thirty-day procedure-related mortality rates were highest for surgery (2.5% vs 0.3% for radiologic gastrostomy and 0.53% for PEG, P < .001). CONCLUSION: Radiologic gastrostomy is associated with a higher success rate than is PEG and less morbidity than either PEG or surgery. PMID- 7480743 TI - Lymph-node metastases: efficacy for detection with helical CT in patients with gastric cancer. AB - PURPOSE: To determine the efficacy of helical computed tomography (CT) in the detection of lymphadenopathy in patients with gastric cancer. MATERIALS AND METHODS: Helical CT (5-mm section thickness, 5-mm/sec table incrementation) was performed in 58 patients (39 men, 19 women; mean age, 63.8 years) with gastric cancer; 1,082 lymph nodes (138 positive, 944 negative for metastasis) were resected at surgery. Findings at CT and resection were compared. Sensitivity for detecting lymph nodes was evaluated according to nodal size and presence of metastasis. RESULTS: Seven (1.1%) of 649 lymph nodes 1-4 mm, 160 (45.1%) of 355 nodes 5-9 mm, and 56 (72%) of 78 nodes larger than 9 mm were detected at CT. For nodes of at least 5 mm, sensitivity for detecting metastasis-positive nodes (79 [75.2%] of 105 nodes) was higher than that for detecting metastasis-negative nodes (137 [41.8%] of 328; P < .01). Differences between positive and negative nodes in CT attenuation (110 HU +/- 25 vs 66 HU +/- 32) and short-to-long axis ratios (0.81 +/- 0.15 vs 0.57 +/- 0.15; P < .001) were significant. CONCLUSION: Helical CT is effective for detection of metastatic lymphadenopathy from gastric cancer. CT attenuation and lymph-node configuration aid in diagnosis of malignant adenopathy. PMID- 7480744 TI - HIV infection: analysis in 259 consecutive patients with abnormal abdominal CT findings. AB - PURPOSE: To evaluate abdominal computed tomographic (CT) findings in patients with human immunodeficiency virus (HIV) infection. MATERIALS AND METHODS: CT scans with abnormal findings in 259 patients (247 men, 12 women; age range, 21-60 years) with HIV infection were analyzed. Diagnoses were mycobacterial infection (n = 87), lymphoproliferative disease (n = 63), Kaposi sarcoma (n = 17), fungal infection (n = 17), hepatocellular disease (n = 13), Pneumocystis carinii infection (n = 8), other disorders (n = 39), or unknown (n = 30). RESULTS: Abnormal findings included lymph-node enlargement (n = 159), hepatomegaly (n = 100), splenomegaly (n = 62), gastrointestinal mass or wall thickening (n = 61), and low-attenuation lesions in the liver (n = 50) or spleen (n = 55). Diagnoses thought to account for CT findings were made (n = 229) or suspected (n = 18) in 247 (95%) of the 259 patients. CONCLUSION: In most patients with HIV infection, abnormal abdominal CT findings are due to a second disease. Even relatively nonspecific findings should not be ascribed to HIV infection and should prompt a search for an opportunistic infection or neoplasm. PMID- 7480746 TI - Small or absent fetal stomach: prognostic significance. AB - PURPOSE: To investigate the outcome of fetuses with a small or absent stomach. MATERIALS AND METHODS: We retrospectively reviewed sonograms in 87 fetuses with a small or absent stomach. Gestational age, amniotic fluid volume, stomach size, morphologic defects, and karyotypes were correlated with clinical follow-up or autopsy findings. RESULTS: Eight pregnancies were excluded because of oligohydramnios due to ruptured membranes. Seventy-nine fetuses with an absent (n = 27) or small (n = 52) stomach were included in the study group. Gestational ages ranged from 18 to 39 weeks (mean, 27 weeks). An abnormal outcome (structural abnormalities, intrauterine fetal or postnatal death) occurred in 23 (85%) of 27 fetuses with an absent stomach and 27 (52%) of 52 fetuses with a small stomach (combined, 63%). Karyotype was abnormal in eight (38%) of 21 fetuses with an absent stomach and two (4%) of 46 fetuses with a small stomach. CONCLUSION: An absent or small fetal stomach after 18 weeks gestation is associated with a guarded prognosis. PMID- 7480745 TI - Microabscesses of the liver and spleen in AIDS: detection with 5-MHz sonography. AB - PURPOSE: To determine whether sonograms of the liver and spleen, obtained with 5 MHz linear-array transducers, aid in detection of hepatosplenic microabscesses in patients with acquired immunodeficiency syndrome (AIDS). MATERIALS AND METHODS: Abdominal sonographic examinations (n = 111) were performed in 102 consecutive patients with AIDS. A 3.5-MHz sector transducer was used in each study, with additional images of the hepatic and splenic parenchyma obtained with a 5-MHz linear-array transducer. Each study was reviewed without benefit of the 5-MHz images, and categories of the hepatic and splenic parenchyma were as follows: 1, lesions definitely present; 2, lesions possibly present; and 3, lesions absent. The 5-MHz images were subsequently reviewed, and studies were recategorized. Findings were correlated with results of pathologic examination. RESULTS: The 5 MHz scans enabled identification of focal hepatic or splenic lesions in 14 of 96 studies placed in category 2 or 3 on the basis of the 3.5-MHz sector scans. CONCLUSION: In patients with AIDS, 5-MHz sonograms of the liver and spleen enable detection of microabscesses not confidently identified on 3.5-MHz scans. PMID- 7480747 TI - Recurrent carcinoma after breast conservation: diagnosis with stereotaxic core biopsy. AB - PURPOSE: To evaluate the utility of stereotaxic core biopsy (SCB) in diagnosing recurrent carcinoma after breast-conserving therapy (BCT). MATERIALS AND METHODS: Of 316 patients evaluated with SCB, 17 (5%) had nonpalpable lesions in breasts previously treated with BCT. Surgical correlation was obtained in 14 patients. SCBs were performed with digital stereotaxic equipment, with patients prone. RESULTS: Carcinoma detected at SCB in 11 (79%) of 14 patients included infiltrating ductal carcinoma (IFDC) in five, ductal carcinoma in situ (DCIS) in five, and infiltrating lobular carcinoma (ILC) in one. Surgical histopathologic findings agreed with core biopsy findings in 10 of these patients. In one patient with two SCB-proved foci of IFDC, one IFDC was identified at mastectomy. In two (14%) patients, SCB revealed atypical ductal hyperplasia; however, DCIS was found at surgical biopsy. In one patient, the SCB finding of fat necrosis was confirmed at surgical biopsy. CONCLUSION: These data suggest that SCB may be useful in diagnosing recurrent carcinoma in the conservatively treated breast. PMID- 7480748 TI - Automated large-core needle biopsy of surgically removed breast lesions: comparison of samples obtained with 14-, 16-, and 18-gauge needles. AB - PURPOSE: To determine the needle size that would consistently enable the pathologist to correctly diagnose tissue core biopsy specimens. MATERIALS AND METHODS: Fifty-seven surgically removed mass lesions of the breast were sampled with a short-throw automated biopsy gun and an 18-gauge needle first, followed by a 16- and a 14-gauge needle. Samples were evaluated independently by three pathologists, and findings were compared with the final diagnosis made at surgical excision biopsy. RESULTS: The sensitivity for the diagnosis of all 26 malignant lesions was 100%, 92%, and 65% for biopsy samples obtained with 14-, 16 , and 18-gauge needles, respectively. Specificity for malignancy was 100% for all three needle sizes. All of the benign diseases were correctly diagnosed in samples obtained with all three size needles except for two samples that did not survive processing. CONCLUSION: Biopsy samples obtained with a 14-gauge needle provide the most accurate diagnosis, which correlates with the diagnosis made with the surgical excision biopsy technique. PMID- 7480749 TI - Breast tumors: comparative accuracy of MR imaging relative to mammography and US for demonstrating extent. AB - PURPOSE: To evaluate the comparative accuracy of magnetic resonance (MR) imaging relative to mammography and ultrasonography (US) for assessing the extent of breast tumors. MATERIALS AND METHODS: Histologic results and preoperative imaging findings (mammography, US, MR imaging) were analyzed regarding tumor size and multifocality of 61 tumors in 60 women undergoing mastectomy for carcinoma. RESULTS: In 10% of cases, the index tumor was not seen at mammography. With US, 15% of the index tumors were not recognized, while MR imaging missed 2% of the index tumors. On mammographic and US images, tumor size was underestimated significantly (P < .005), by 14% and 18%, respectively, while MR imaging showed no significant difference in size compared with that found in a pathologic evaluation. Mammography showed 31% of the additional invasive lesions, while US showed 38% and MR imaging showed 100%. CONCLUSION: MR imaging was the most accurate of the three preoperative imaging modalities in assessing the size and number of malignant lesions in the breast. PMID- 7480750 TI - Scanner conformity in CT densitometry of the lungs. AB - PURPOSE: To quantify inter- and intrascanner conformity in computed tomographic (CT) densitometry of the lungs. MATERIALS AND METHODS: With six scanners from four manufacturers, a lung densitometry protocol with several variations was applied for performance comparison. Phantoms included water, air, and a humanoid thorax phantom equipped with a dog lung and exchangeable pseudolungs of polyethylene foam. RESULTS: All scanners produced acceptable CT numbers (Hounsfield units) for water, but some not for air. An incorrect calibration of air density affected all CT numbers at lung densities, but the error was easily correctable. Some systems were more sensitive to object size than others were. Sensitivity of CT numbers to section thickness, reconstruction filter, zoom factor, and table height was small, except for two scanners in relation to section thickness. CONCLUSION: After correction for poor air calibration, scanner conformity was acceptable when the reproducibility of lung densitometry in clinical practice was set as a reference. PMID- 7480751 TI - Interpretation time of serial chest CT examinations with stacked-metaphor workstation versus film alternator. AB - PURPOSE: Interpretation time of serial staging chest CT cases, which each contained current and previous examinations, with a simple prototype workstation called filmstack was experimentally compared with interpretation time with a film alternator. MATERIALS AND METHODS: The filmstack displayed a "stack" of sections for each examination; user controls allowed rapid selection of preset attenuation windows and both synchronized and unsynchronized scrolling. Eight radiologists were timed as they used the filmstack and the film alternator to interpret four ergonomically complex serial CT cases. RESULTS: All reports dictated on the basis of findings with filmstack and film were of acceptable clinical accuracy. The time to examine a case with filmstack was significantly faster than the time with film, including the time to load and unload the alternator (99% confidence [P = .01]). There was no statistically significant difference in interpretation time between filmstack and prehung film. CONCLUSION: Use of a low-cost stacked CT workstation with a single 1,024 x 1,024 monitor is an effective means of interpreting cases that require comparison of multiple CT examinations. PMID- 7480752 TI - Added value of radiologist consultation to family practitioners in the outpatient setting. AB - PURPOSE: To measure the added value of a radiologist's consultation to the interpretation of radiographs previously read by a family practitioner. MATERIALS AND METHODS: The authors reviewed 1,674 chest and extremity radiographs previously read by a family practitioner and consulting radiologist. The 196 radiographs in which there was a discrepancy between the family practitioner's and radiologist's report were evaluated by a radiologist and family physician not involved in and blinded to the original interpretations. The overall accuracy of the participants was determined and differences statistically quantified. RESULTS: The overall sensitivity of the radiologists was greater than that of the family practitioners (92% vs 86%); specificity was not significantly different. For extremity examinations, there were no significant differences in accuracy of the radiologists and family practitioners; the sensitivity of radiologists for chest studies was considerably greater (89% vs 80%). Radiologic consultation was of particular value in the detection of pneumonia and masses. CONCLUSION: At a family practice center, the radiologist's role for extremity radiographs might be limited to individual consultation, with review of all chest radiographs. PMID- 7480753 TI - Fractures of the first rib in child abuse. AB - PURPOSE: To evaluate the incidence and specificity of fracture of the first rib as an indicator of child abuse and to determine a mechanism of fracture. MATERIALS AND METHODS: All infants and young children with rib fractures who were seen at the radiology department of an academic medical center over a 2-year period were identified from the radiology data base. RESULTS: Rib fractures were attributed to child abuse in 12 of 35 children identified. Apart from a neonate with congenital osteogenesis imperfecta, only three children, all abused, had first-rib fracture, one bilaterally. Two additional children with first-rib fracture were identified from years prior to the study period. In four children, first-rib fractures were "isolated," without fractures of adjacent bones. CONCLUSION: Child abuse should be considered in cases of pediatric rib fracture, particularly fracture of the first rib. Possible mechanisms for first-rib fracture include impact force, compressive force, and shaking or acute axial load (slamming), which cause an indirect fracture. PMID- 7480754 TI - Cartilaginous abnormalities and growth disturbances in Legg-Calve-Perthes disease: evaluation with MR imaging. AB - PURPOSE: To evaluate the magnetic resonance (MR) imaging appearance of cartilaginous abnormalities in Legg-Calve-Perthes (LCP) disease and their association with growth arrest. MATERIALS AND METHODS: Twenty-seven MR images and radiographs of 25 hips in 23 patients were reviewed for epiphyseal, physeal, and metaphyseal abnormalities. Follow-up radiographs (mean time, 20.5 months) were evaluated for growth arrest. RESULTS: Epiphyseal abnormality seen in 74% of cases was not related to growth disturbances. Transphyseal bone bridging on MR images, the strongest predictor of abnormal growth (P < .01), was found in 63% of femurs. MR imaging helped detect physeal abnormalities with greater sensitivity (94%), specificity (89%), and positive predictive value (94%) than radiography. Metaphyseal extension of physeal cartilage seen in 81% of cases was associated with growth abnormality (P < .01), particularly when detected at radiography. CONCLUSION: Cartilaginous physeal and metaphyseal abnormalities in LCP disease are common and frequently result in growth arrest. PMID- 7480755 TI - Peripherally inserted central catheters in children. AB - PURPOSE: To assess the feasibility and complications of peripherally inserted central catheters (PICCs) in pediatric patients. MATERIALS AND METHODS: The authors attempted to place PICCs in 122 patients aged 9 days to 19 years (mean, 6.82 years; median, 5 years). Catheters were placed to allow prolonged administration of antibiotics or chemotherapeutic agents (n = 50), provide total parenteral nutrition (n = 41), and establish prolonged intravenous access for blood draws and fluid administration (n = 31). Silicone catheters measuring 3, 4, and 5 F were inserted in either basilic or cephalic veins and positioned at the junction of the superior vena cava and right atrium under fluoroscopic guidance. Patients were monitored for complications until devices were removed. RESULTS: Fluoroscopically guided PICC placement was successful in 137 of 148 attempts. Postinsertion complications included mechanical defects of the catheter, PICC related infection, occlusion of the PICC, and venous stasis. Complications occurred at a rate comparable to those seen with blind insertion. CONCLUSION: Fluoroscopically guided PICC placement is feasible and safe in pediatric patients. PMID- 7480756 TI - Calcification of coronary arteries: detection and quantification with double helix CT. AB - PURPOSE: To evaluate the accuracy of double-helix computed tomography (CT) in coronary artery calcification detection and quantification. MATERIALS AND METHODS: One hundred sixty patients with coronary disease (135 men, 25 women; age range, 45-62 years), of whom 138 had obstructive (stenosis of > 50% of diameter; n = 129) or mild (< 50% stenosis; n = 9) coronary artery disease (CAD) and 22 had normal coronary arteries (per angiographic findings), and 56 age-matched healthy control subjects underwent double-helix CT. RESULTS: Double-helix CT findings indicated that calcification was significantly more prevalent in patients with CAD (> 83%) than in patients with normal coronary arteries (27%) or in healthy control subjects (34%; P < .01). Sensitivity in detecting obstructive CAD was high (91%); however, specificity was low (52%) because of calcification in nonobstructive lesions. Comparison of double-helix CT and angiographic findings indicated that double-helix CT was 84% accurate with positive and negative predictive values of 89% and 59%, respectively. CONCLUSION: Double-helix CT is a useful noninvasive method for detection and quantification of coronary artery calcification. PMID- 7480758 TI - Renal arteries: clinical comparison of three-dimensional time-of-flight MR angiographic sequences and radiographic angiography. AB - PURPOSE: To prospectively compare three magnetic resonance (MR) angiographic techniques in the renal arteries. MATERIALS AND METHODS: Twenty-five adult patients underwent three-dimensional time-of-flight MR angiography with three different sequences: conventional, tilted optimized nonsaturating excitation (TONE), and selective inversion-recovery rapid gradient-echo (SIR-RAGE). Fifteen also underwent radiographic angiography. Stenosis grade measured with each MR angiographic technique was compared with that measured with radiographic angiography by using correlation coefficients. Visible artery lengths with each MR angiographic technique were compared by using the Turkey method. RESULTS: Correlation between stenosis grades with each MR angiographic technique and with radiographic angiography was good (P < .01). Stenosis was correctly excluded with SIR-RAGE findings in six patients. Mean visible artery length was greatest with SIR-RAGE (P < .01). CONCLUSION: SIR-RAGE depicts a greater length of renal artery than does conventional MR angiography. TONE also improved depiction of distal arteries. Normal SIR-RAGE findings were highly predictive of normal arteries. PMID- 7480759 TI - Percutaneous endoluminal placement of stent-grafts for the treatment of isolated iliac artery aneurysms. AB - PURPOSE: To evaluate the feasibility of stent-grafts for treatment of isolated iliac artery aneurysms (IAAs). MATERIALS AND METHODS: Nine IAAs in eight patients were treated with transluminally placed endovascular stent-grafts. All patients were men (median age, 72 years). In three, the aneurysm involved both the common and internal iliac arteries. In one, common and external iliac arteries were involved. The other aneurysms involved only the common iliac artery. Two aneurysms were treated with balloon-expandable stents covered with polytetrafluoroethylene (PTFE) graft material, three were treated with self expanding Z-stents covered with a woven polyester graft, and four were treated with self-expanding Z-stents covered with PTFE. RESULTS: Transluminal placement of the stents was successful in all patients with thrombosis of the aneurysms. There were no distal thromboembolic events, deaths, or infections. The median follow-up period was 8.5 months. CONCLUSION: Initial results suggest that transluminal stent-graft placement for treating isolated IAAs is a safe and effective alternative to surgery in selected patients. Long-term follow-up data are needed before this approach can be recommended for the primary treatment of IAAs. PMID- 7480757 TI - Breath-hold gadolinium-enhanced MR angiography of the abdominal aorta and its major branches. AB - PURPOSE: To develop and evaluate a sequence for breath-hold three-dimensional gadolinium-enhanced magnetic resonance (MR) angiography of the abdominal aorta. MATERIALS AND METHODS: In 63 patients, the abdominal aorta and its branches were imaged for 29, 43, or 58 seconds with breath holding. A fast spoiled gradient echo sequence was used at 1.5 T during infusion of 42 mL of a gadolinium chelate. Correlation with conventional angiography was performed in 19 patients. MR image quality (signal-to-"total" noise ratio [S/N*]) with breath holding was compared with that with free breathing (104 patients). RESULTS: With breath-hold gadolinium-enhanced MR angiography, renal, celiac, and superior mesenteric artery occlusive disease was graded appropriately in 15 of 19 patients, and 10 of 11 accessory renal arteries were depicted correctly. Renal artery branches were visualized in 86 of 95 kidneys on breath-hold images compared with only 84 of 236 kidneys with free breathing (P < .001). Distal renal artery S/N* was 3.1 with breath holding and 2.1 with free breathing (P < .001). CONCLUSION: Breath holding statistically significantly improves three-dimensional gadolinium-enhanced MR angiography of the renal, celiac, and superior mesenteric arteries. PMID- 7480760 TI - Budd-Chiari syndrome: technical, hemodynamic, and clinical results of treatment with transjugular intrahepatic portosystemic shunt. AB - PURPOSE: To evaluate use of the transjugular intrahepatic portosystemic shunt (TIPS) as a nonsurgical approach for the management of Budd-Chiari syndrome (BCS). MATERIALS AND METHODS: Twelve patients with fulminant (n = 2), subacute (n = 5), or chronic (n = 5) BCS underwent TIPS placement. Hepatic venous obstruction was demonstrated at computed tomography and color duplex sonography. BCS was confirmed histologically in all patients. Hemodynamic parameters and clinical characteristics were assessed. RESULTS: TIPS creation was successful in all patients. Treatment reduced the portal venous pressure gradient by 75% and resulted in a mean shunt flow of 2,300 mL/min +/- 650 (standard deviation). No serious procedure-related complications were observed. The two patients with fulminant BCS died of septicemia or progressive liver failure despite intervention. The other 10 patients showed clinical improvement with reduction or disappearance of ascites. During follow-up, shunt dysfunction occurred in five of 10 patients with recurrence of ascites requiring repeat intervention. CONCLUSION: TIPS placement is safe and effective in patients with portal hypertension caused by subacute or chronic BCS. PMID- 7480761 TI - Treatment of large HCC: transcatheter arterial chemoembolization combined with percutaneous ethanol injection versus repeated transcatheter arterial chemoembolization. AB - PURPOSE: To compare the efficacy of transcatheter arterial chemoembolization (TACE) combined with percutaneous ethanol injection (PEI) versus repeated TACE in the treatment of large hepatocellular carcinoma (HCC). MATERIALS AND METHODS: Fifty-three patients with cirrhosis and a large HCC (main tumor, 3.1-8.0 cm in diameter with no more than two daughter nodules) were enrolled in a prospective, randomized study. Twenty-six patients underwent a single TACE session followed by PEI (TACE-PEI group), whereas 27 patients underwent two to five TACE sessions (TACE group). Both groups of patients were similar with regard to liver function. Follow-up ranged from 8 to 39 months. RESULTS: Complete therapeutic responses were higher (P < .05) and tumor recurrences during follow-up were lower (P < .05) in the TACE-PEI group than in the TACE group. Patients in the TACE-PEI group survived longer than those in the TACE group, although the difference was not significant (P > .1). The rates of survival without recurrence were better in the TACE-PEI group than in the TACE group (P < .05). CONCLUSION: Use of a single TACE session combined with PEI is more effective than repeated TACE in the treatment of large HCC. PMID- 7480762 TI - Anterior shoulder instability: diagnostic criteria determined from prospective analysis of 121 MR arthrograms. AB - PURPOSE: To determine magnetic resonance (MR) arthrographic criteria in the diagnosis of anterior glenohumeral instability. MATERIALS AND METHODS: In 121 patients with diagnoses proved surgically, the labrum, glenohumeral ligaments, and capsular insertion types were assessed prospectively with gadolinium-enhanced MR arthrography. Findings were compared in stable and unstable shoulders. RESULTS: Operative results showed 59 normal, 57 torn, and five deficient labra. In 37 unstable shoulders, 31 had discrete inferior labral-ligamentous lesions and six had capsular laxity. MR arthrograms showed labral abnormalities with 92% sensitivity, 92% specificity. Inferior labral-ligamentous lesions enabled prediction of anterior instability with 76% sensitivity (capsular laxity was missed in all shoulders), 98% specificity. Inferior labral-ligamentous abnormalities were strongly associated with unstable shoulders (P << .0001), whereas noninferior labral-ligamentous abnormalities were related to stable shoulders (P = .01). Capsular insertion types showed no significant differences between stable and unstable shoulders (P > .8). CONCLUSION: On MR arthrograms, inferior labral-ligamentous abnormalities were most closely correlated with anterior glenohumeral instability. Capsular insertion sites had no role in the prediction of shoulder instability. PMID- 7480763 TI - Anterior cruciate ligament tear: prospective evaluation of diagnostic accuracy of middle- and high-field-strength MR imaging at 1.5 and 0.5 T. AB - PURPOSE: To compare the diagnostic efficacy of current-generation middle- and high-field-strength magnetic resonance (MR) imagers in the diagnosis of anterior cruciate ligament (ACL) tears. MATERIALS AND METHODS: In 114 of 230 patients referred for knee imaging, MR imaging at 0.5 and 1.5 T was performed with identical sequences but with a slightly longer total imaging time and bandwidth optimization at 0.5 T. Radiologists were blinded to diagnosis and field strength. Sensitivity, specificity, and accuracy were determined, and ACL tear was confirmed by means of arthroscopy and pathology. RESULTS: There was no difference between the field strengths in accuracy, sensitivity, or specificity for the diagnosis of ACL tears in 86 patients with disrupted ACLs and 28 patients with intact ACLs. Accuracy for all ACL tears was 90% at 0.5 T and 91% at 1.5 T. Similarly, there were no differences in diagnosis of meniscal tears (79 with, 149 without) or posterior cruciate ligament tears (seven with, 107 without). CONCLUSION: Higher field strength does not confer higher accuracy in the diagnosis of ACL tears at MR imaging. PMID- 7480765 TI - Study of osteomyelitis: utility of combined histologic and microbiologic evaluation of percutaneous biopsy samples. AB - PURPOSE: To determine the utility of combined histologic and microbiologic evaluation of percutaneous biopsy samples in cases of suspected osteomyelitis. MATERIALS AND METHODS: Twenty-five patients with suspected osteomyelitis were prospectively studied over a 1-year period. With usual trephine techniques under radiologic guidance, core and aspiration biopsy samples were obtained in each patient for both histologic and microbiologic analyses. RESULTS: Sixteen biopsy specimens demonstrated histologic evidence of osteomyelitis. Of these, eight were also culture positive. Seven of the eight culture-negative, histologically positive cases were interpreted as chronic osteomyelitis. In no patient were cultures positive and histologic findings negative. Three of 19 patients with proved osteomyelitis had negative histologic and microbiologic findings. The sensitivity of culture in the diagnosis of osteomyelitis in our study was 42%; the sensitivity of both culture and histologic findings was 84%. CONCLUSION: Although the volume of a biopsy specimen is a major determinant of culture yield, a portion of a percutaneous biopsy sample should be histologically evaluated for possible osteomyelitis. PMID- 7480764 TI - Musculoskeletal neoplasm: perineoplastic edema versus tumor on dynamic postcontrast MR images with spatial mapping of instantaneous enhancement rates. AB - PURPOSE: To evaluate the utility of fast, contrast-enhanced, sequential magnetic resonance (MR) imaging in differentiating between extraosseous tumor and perineoplastic edema. MATERIALS AND METHODS: Fourteen patients underwent sequential MR imaging (3.5 seconds per image) after bolus administration of gadopentetate dimeglumine. Initial rates of enhancement (initial slope) were calculated on a pixel-by-pixel basis and displayed as a "slope image"' in which pixel intensity reflected the slope value. Close correlation with wedge biopsy specimens was performed. RESULTS: Mean initial slope values were viable extraosseous tumor, 9.33 (standard deviation, 2.23); infiltrated muscle, 9.07 (2.31); edematous muscle without tumor infiltration, 5.48 (1.27); normal muscle, 1.11 (0.65). Differences in initial slope between all neoplastic and nonneoplastic tissues were statistically significant. Within individual patients, initial slope of edematous muscle was always 20% or more lower than that of neoplastic tissue. Slope images highlighted areas of viable extraosseous tumor and infiltrated muscle against edematous and normal tissues. CONCLUSION: Computer generated slope images derived from sequential postcontrast MR images allow differentiation between tumor and nonneoplastic edema and may thereby guide the surgeon in planning limb-sparing procedures. PMID- 7480767 TI - CT bone window photography in patients with cancer. AB - PURPOSE: To assess the utility of routinely photographing computed tomographic (CT) bone windows in patients with cancer. MATERIALS AND METHODS: The impression section of body CT reports were reviewed in 4,683 patients with cancer (2,240 female and 2,443 male patients, aged 2 months to 97 years [mean, 55 years]). RESULTS: The presence of definite or possible bone metastasis was mentioned in 523 (11.2%) patients. No prior radiologic examination was available in 165 patients. In the 358 patients who had undergone previous radiologic examinations, findings were positive in 271 and normal or indeterminate in 87. In 252 (5.4%) patients, CT with bone windows may have been needed for diagnosis of bone metastasis: 110 had extensive nonosseous metastases, 77 had no osseous metastasis, 19 had bone findings that were not followed up radiologically, and 46 had bone lesions that were new findings. These new lesions were visible on scans photographed at soft-tissue windows in 45 (97.8%) patients. CONCLUSION: Routine photography of CT bone windows is not necessary in patients with cancer. PMID- 7480766 TI - Patellar tendinitis: MR imaging features, with suggested pathogenesis and proposed classification. AB - PURPOSE: To characterize the magnetic resonance (MR) imaging features of patellar tendinitis. MATERIALS AND METHODS: Fifteen patients with a clinical diagnosis of patellar tendinitis underwent gadolinium-enhanced MR imaging of the knee. RESULTS: Grades of patellar abnormality, based on findings in the enthesial region at MR imaging, correlated with signs of increasing fibrovascular repair: grade 1 (n = 4), enhancing area adjacent to patellar apex, with marginal zone of intermediate signal intensity, and a patellar apical chondral-bone avulsion; grade 2 (n = 5), same signs as grade 1 damage but without avulsion; grade 3 (n = 6), homogeneous, nonenhancing area of intermediate signal intensity adjacent to the patellar apex seen on all images. Changes were most obvious posteriorly and involved the central and medial thirds of the tendon. Chronic injury to the medial retinaculum was a common associated finding. CONCLUSION: Patellar tendinitis demonstrates a consistent spectrum of changes at MR imaging that can aid understanding of the origin and treatment of damage. PMID- 7480768 TI - Iliopsoas bursa imaging: efficacy in depicting abnormal iliopsoas tendon motion in patients with internal snapping hip syndrome. AB - PURPOSE: To evaluate the efficacy of both iliopsoas bursa imaging and therapeutic injections for determination of the cause of pain in patients with internal snapping hip syndrome. MATERIALS AND METHODS: Twelve patients underwent iliopsoas bursa imaging. One patient underwent the procedure bilaterally and five underwent repeated studies for a total of 21 studies. Eight patients also received therapeutic injections after bursa imaging. RESULTS: Abnormal motion of the iliopsoas musculotendinous unit that occurred synchronously with the snap was revealed in 15 of the 21 studies. Four studies revealed additional information that contributed to symptoms. Eight patients received therapeutic injections with relief of symptoms that ranged from 2 weeks to 2 years. CONCLUSION: Iliopsoas bursa imaging is a simple, rapid, and reproducible method with which abnormal iliopsoas tendon motion can be documented in patients with internal snapping hip syndrome and is also useful for diagnosis of other causes of atypical hip pain. Therapeutic injection into the iliopsoas bursa may delay or obviate surgery. PMID- 7480769 TI - Report from the First Roentgen Centennial Fellow. PMID- 7480770 TI - Limited-stage small-cell lung cancer: patient survival after combined chemotherapy and radiation therapy with and without treatment protocols. AB - PURPOSE: To compare survival of patients who undergo treatment in protocols versus survival of patients not in protocols. MATERIALS AND METHODS: Records of 81 adult patients with small-cell lung cancer who underwent chemotherapy and radiation therapy in 1987-1992 were reviewed retrospectively. Forty-one patients were in a protocol; 40 patients were not. Patient demographics and prognostic factors were not statistically significantly different. RESULTS: Median overall survival was 16.7 months in the nonprotocol group versus 29.0 months in the protocol group (P = .0023). Median disease-specific survival was 18.3 months in the nonprotocol group versus 27.1 months in the protocol group (P = .0176). Survival was not statistically significantly influenced by Karnofsky performance status, weight loss, or thoracic radiation dose. CONCLUSION: There was a highly statistically significant difference in survival outcome in the nonprotocol group versus the protocol group (P = .0023). Differences in chemotherapy-radiation therapy timing and other treatment-related factors may have contributed substantially to the improved survival in the protocol group. PMID- 7480771 TI - Decreased acute toxicity by using midline mucosa-sparing blocks during radiation therapy for carcinoma of the oral cavity, oropharynx, and nasopharynx. AB - PURPOSE: To determine whether midline mucosa-sparing blocks (MSBs) protecting the aerodigestive tract can statistically significantly reduce acute toxicity during radiation therapy for carcinoma of the head and neck, without compromising tumor control. MATERIALS AND METHODS: Radiation records and simulation films were reviewed in 125 patients with carcinoma of the oral cavity, oropharynx, or nasopharynx. Patients with and without MSBs were compared. Measures of acute toxicity during radiation therapy were weight loss (> or = 5%), hospitalization for nutritional support, and unplanned treatment interruptions (> or = 5 days). Actuarial local-regional tumor control was compared. RESULTS: Patients with MSBs had significantly less weight loss (26 of 50 vs 37 of 47 patients, P = .006), fewer hospitalizations for nutritional support (one of 61 vs seven of 64 patients, P = .04), and a trend toward fewer treatment interruptions (10 of 61 vs 19 of 64 patients, P = .07) than patients without MSBs. The 3-year actuarial tumor control rates in the neck were similar. CONCLUSION: Midline MSBs decrease acute toxicity during radiation therapy for carcinoma of the oral cavity, oropharynx, and nasopharynx without compromising tumor control. PMID- 7480772 TI - Intracranial aneurysms: endovascular evaluation with three-dimensional-display MR angiography. AB - To augment information at digital subtraction angiography, 17 patients with 22 intracranial aneurysms underwent three-dimensional-display magnetic resonance (MR) angiography. Reconstructions created to simulate vascular endoscopy provided additional information in 18 aneurysms about the aneurysmal sac, its orifice and neck, and/or its relation to the parent vessel and nearby vasculature. No additional information was provided in two aneurysms, and two aneurysms were not depicted. PMID- 7480773 TI - "Failure to correctly diagnose" is not violation of EMTALA. PMID- 7480775 TI - GA: is there a nurse-patient privilege?: caveat before disclosing information. PMID- 7480776 TI - LA: E.R. treatment delayed 2 1/2 hours: delay not proximate cause of death. PMID- 7480774 TI - Nurses fail to monitor blood sugar despite family pleas. PMID- 7480777 TI - Charge nurse prevents "call to doctor": catastrophic results. PMID- 7480778 TI - Nurse risk manager alleges "retaliatory transfer". PMID- 7480781 TI - OH: "LifeFlight" nurse & pilot to marry: hospital's nepotism policy mandates transfer. PMID- 7480780 TI - MI: independent living home pt. drowns: failure to follow nursing care plan. PMID- 7480779 TI - Arbitrator's award of E.R. job to existing employee upheld. PMID- 7480782 TI - Pt. allergic to meds: nurse placing hep-lock joined as defendant. PMID- 7480783 TI - Colloidal gold post-embedding immunocytochemistry. PMID- 7480784 TI - Kainic acid seizures in the rat. PMID- 7480785 TI - Nitric oxide: a radical neurotransmitter in the central nervous system. PMID- 7480786 TI - Endogenous control of epilepsy: the nigral inhibitory system. PMID- 7480788 TI - The structure and function of Pacinian corpuscles: a review. PMID- 7480789 TI - [Development of information biology]. PMID- 7480787 TI - Monoamine oxidase: distribution in the cat brain studied by enzyme- and immunohistochemistry: recent progress. AB - Localization of MAO-containing neurons, fibers and glial cells has been described by recent progress in MAO histochemistry and immunohistochemistry. It does not necessarily correspond to those containing monoamines. MAO-A is demonstrated in many noradrenergic cells, but it is hardly detectable in DA cells. Increase of 5 HT and DA concentration after inhibition of MAO-A indicates the possible existence of MAO-A in such neuronal structures. MAO-A is also undetectable in neurons containing 5-HT, a good substrate for MAO-A. These neurons contain MAO-B. There still remain contradictions to be solved in future. MAO is present in astroglial cells, in which monoamines released in extracellular space may be degraded. In glial cells, MAO may also play a role to regulate concentration of telemethylhistamine and trace amines. Such cells appear to transform MPTP to MPP+, a neurotoxin for nigral DA neurons. PMID- 7480790 TI - [Toward virtual life from bioinformatics]. PMID- 7480791 TI - [Epstein-Barr virus oncogenesis: immortalization by a human ubiquitous virus and neoplastic conversion by risk factors]. PMID- 7480792 TI - [The catalytic mechanism of carbonic anhydrase]. PMID- 7480793 TI - [Taste sensor]. PMID- 7480795 TI - [South-Hyougo earthquake and Institute for Protein Research]. PMID- 7480794 TI - [An ambient and low temperature two-dimensional electrophoresis: a convenient way to detect cold-dissociable proteins]. PMID- 7480796 TI - Prostaglandin E2 regulates inducible nitric oxide synthase in the murine macrophage cell line J774. AB - We have evaluated the role of prostaglandin E2 (PGE2) in the synthesis of nitric oxide (NO) by the activation of the inducible form of nitric oxide synthase (NOS) in the murine macrophage cell line, J774, stimulated with different doses of lipopolysaccharide (LPS). The stimulation of the J774 line with suboptimal doses of LPS (0.1 microgram/mL) caused a production of endogenous PGE2 that was capable of stimulating NOS activity inducing an increase in the NO synthesis, as attested by the fact that cyclooxygenase enzyme inhibitor, indomethacin, significantly reduced NO secretion. On the contrary, a higher dose of LPS (1 microgram/mL) produced high levels of PGE2 that reduced the levels of NOS and, subsequently, NO production. Experiments carried out with exogenous PGE2 indicated that concentrations between 1 and 10 ng/mL are able to stimulate the expression of NOS and the release of NO, while higher concentrations (> 50 ng/mL) are inhibitory. Furthermore, our data indicate that there is a network of interaction which involves NO, PGE2, and tumor necrosis factor. High levels of PGE2 inhibited TNF alpha secretion, which in turn could exert inhibitory effects on NO synthesis. PMID- 7480797 TI - Leukotriene C4-stimulated contractions in bullfrog lung are affected by cold acclimation and calcium antagonists. AB - Leukotriene C4 (LTC4) contracts isolated bullfrog lung. This study examined effects of cold-acclimation and the involvement of extracellular and intracellular Ca++ on the contractile response to LTC4. The response to LTC4 was greater in lungs from warm-acclimated (22 degrees C) frogs compared with cold acclimated (5 degrees C) frogs at incubation temperatures of both 22 degrees C and 5 degrees C. LTC4, LTC5, and N-methyl LTC4 were equally effective in stimulating lung contraction at concentrations from 1-100 nM. Nicardipine (3 microM) partially antagonized the response to LTC4, but verapamil, nifedipine, or nitrendipine at the same concentration was ineffective. Ethylene glycol tetraacetic acid (EGTA, 0.3 mM) prevented the response to 30 nM LTC4, but the response was restored when the lung was retested in EGTA-free medium containing Ca++, suggesting that extracellular Ca++ was involved in the response. Caffeine (10 mM) or thapsigargin (1 mM) inhibited the responses to LTC4, suggesting a role for intracellular Ca++ in the contraction. PMID- 7480798 TI - Interleukin-1 beta-stimulated prostaglandin synthesis by human decidual cells is independent of protein kinase C. AB - Basal prostaglandin E2 (PGE2) synthesis by human decidual cells was stimulated by phorbol myristate acetate (PMA) which activates protein kinase C. Staurosporine, which is an inhibitor of protein kinase C in most systems, also increased basal PGE2 synthesis. Further work is needed to explain this finding, as another inhibitor of protein kinase C, H7, inhibited PGE2 production under similar culture conditions. Interleukin-1 beta (IL-1 beta)-stimulated PGE2 synthesis was potentiated by coincubation with PMA or staurosporine, indicating that IL-1 beta and protein kinase C increase decidual PGE2 synthesis through different mechanisms. Desensitization of the decidual cells for 24 h with PMA did not affect IL-1 beta-stimulated PGE2 synthesis. The complex roles of protein kinase C in regulating decidual prostaglandin synthesis require further investigation, but it is clear that the effects of IL-1 beta are not mediated by protein kinase C. PMID- 7480799 TI - Induction of alkalinization in cultured renal cells (MDCK line) by prostaglandin E2. AB - The effect of prostaglandin E2 (PGE2), on the intracellular pH (pHi) in BCECF loaded Madin Darby Canine Kidney (MDCK) cells was investigated. PGE2 elevated the pHi. Under resting conditions, pHi of MDCK cells suspended in PBS at pH 7.4 was 7.11 +/- 0.08; PGE2 increased pHi with an EC50 of 0.16 microM. PGF2 alpha elicited a similar response to PGE2, with an EC50 of 0.24 microM. Amiloride (0.4 mM) reversed the response to PGE2 (control 7.18 +/- 0.05; PGE2 7.26 +/- 0.05; after amiloride 7.18 +/- 0.05). In MDCK cells exposed to a Na(+)-free solution, alkalinization induced by this eicosanoid was blocked (Ringer-choline 7.16 +/- 0.03; PGE2 7.16 +/- 0.02). PGE2 increased by 100% the rate of recovery after an acidification pulse with ammonium chloride. In the presence of Ringer-HCO3- (pH 7.4), there was a delay in the maximal response to this prostaglandin (PBS 2.2 +/ 0.27, Ringer-bicarbonate 3.4 +/- 0.55 min) and the pHi increment was less marked than in PBS (0.09 pH units in HCO3- versus 0.16 pH units in PBS; P < 0.001). This effect of PGE2 was not blocked by 4,4'-diisothiocyanatostilbene-2,2'-disulfonic acid (1.0 mM). PMA (100 nM), activator of protein kinase C, mimicked the response to PGE2, suggesting the participation of this kinase on the effect of the prostanoid. As expected, two inhibitors of protein kinase C, staurosporine and sphingosine, abolished the response to PGE2. Staurosporine (0.10 microM), an inhibitor of protein kinase C, blocked the response to PGE2 (control 7.02 +/- 0.04; PGE2 and staurosporine 7.03 +/- 0.04, n = 9, not significant). Sphingosine, another inhibitor of protein kinase C, also blocked the response to PGE2. Two analogues of cAMP did not modify the pHi. In summary, PGE2 induced an intracellular alkalinization via stimulation of a Na+/H+ exchanger, with the participation of protein kinase C, in MDCK cells. PMID- 7480801 TI - Obituary for Professor Priscilla Piper. PMID- 7480802 TI - Oscillating prostacyclin and thromboxane generation by human vessels: biological and mathematical evidence for negative feedback control. AB - The presented study investigates the time-dependent release of PGI2 and TXA2 by isolated human umbilical veins in vitro using the radioimmunoassay for measurement. After changing the nutritional fluid--Krebs-Henseleit solution at pH 7.4, 37 degrees C, 95% O2/5% CO2--the release graph oscillates. These oscillations with time were verified by variance analysis and are very similar for both substances. This indicates one or several negative feedback mechanisms acting on the common path of synthesis from the membrane-bound phospholipids to PGH2, which are effective in the regulation of eicosanoid biosynthesis in vitro. A mathematical function describing the observed PGI2 and TXA2 synthesis is communicated. PMID- 7480800 TI - Beta-endorphin inhibits prostaglandin synthesis in rat ovaries and blocks induced ovulation. AB - The aim of the present study was to explore the action of exogenous beta endorphin on the number of oocytes ovulated and on prostaglandin (PG) production in ovaries isolated from pregnant mare's serum gonadotropin/human chorionic gonadotropin(PMSG/hCG)-primed immature rats. An intrabursal injection of the opioid (0.084 microgram) was given 4 hours after hCG and the number of oocytes within the oviducts on the following morning was reduced (P < 0.05). The same effect was also attained with an intraperitoneal (IP) injection (0.5 microgram). The time course of PG synthesis was quantified in ovaries of rats treated with an IP injection. Eight hours after hCG, prostaglandin content increased (P < 0.01) and remained high until 12 hours after hCG (P < 0.001). This increase was inhibited by the in vivo treatment with beta-endorphin. On isolated ovaries, beta endorphin (10(-8) M) had a clear inhibitory action on prostaglandin production. beta-Endorphin's effect on prostaglandin synthesis in the ovaries is of importance in the ovulatory process. The possible physiological role of beta endorphin merits further investigation. PMID- 7480803 TI - Prevention of abnormalities in motor nerve conduction and nerve blood-flow by a prostacyclin analog, beraprost sodium, in streptozotocin-induced diabetic rats. AB - The effects of the prostacyclin analog beraprost sodium on motor nerve function and nerve blood-flow were examined in streptozotocin-induced diabetic rats. Oral administration of beraprost sodium 0.1 mg/kg/day for 8 wks significantly (P < 0.001) improved caudal motor nerve conduction velocity and sciatic nerve blood flow, both of which are impaired in diabetic rats. Beraprost sodium did not affect glucose, sorbitol, or fructose levels in the sciatic nerve. However, a decreased content of cyclic AMP in the sciatic nerve and higher level of thromboxane B2 in the thoracic aorta of diabetic rats, as compared with those in normal rats, were reversed by the treatment with beraprost sodium (P < 0.01). Results suggest that beraprost sodium may have value in treating diabetic neuropathy, mainly by improving endoneurial blood-flow. PMID- 7480805 TI - Regulation of two isozymes of prostaglandin endoperoxide synthase and thromboxane synthase in human monoblastoid cell line U937. AB - The mechanism responsible for the rapid increase of thromboxane A2 synthesis by cells of the human monoblastoid cell line U937, which were differentiated with 12 O-tetradecanoyl-phorbol-13-acetate, induced by lipopolysaccharide (LPS) was studied. Both RNA blot and immunoblot analyses showed that LPS increased the levels of prostaglandin endoperoxide synthase-1 (PES-1) and -2 (PES-2) in a time dependent manner, and the modes of induction of the two isozymes differed. The maximum PES-1 mRNA level was 1.6 times higher 36 h after than before stimulation by LPS, and that of PES-2 mRNA was elevated about 20-fold at its peak at 12 h after stimulation. Consequently, the immunoreactive PES-1 and PES-2 protein levels also increased time-dependently after LPS stimulation. However, the effects of LPS on the thromboxane synthase mRNA and protein levels were much less marked. These results indicate that LPS-induced thromboxane synthesis by the differentiated cells was regulated at the levels of the two PES isozymes, predominantly at the PES-2 level. PMID- 7480804 TI - Interleukin-1 beta induces the synthesis and activity of cytosolic phospholipase A2 and the release of prostaglandin E2 in human amnion-derived WISH cells. AB - The objective of this study was to examine the expression and activity of cytosolic phospholipase A2 (cPLA2) in relation to prostaglandin E2 (PGE2) synthesis in human amnion-derived WISH cells in response to stimulation by interleukin-1 beta (IL-1 beta). cPLA2 activity was characterized by sensitivity to heat and acid treatment, stability to dithiothreitol, and inhibition by the specific inhibitor, arachidonyl trifluoromethyl ketone (AACOCF3). Treatment of WISH cells with IL-1 beta (0.01-1 ng/mL) for up to 24 h resulted in a significant increase in PGE2 release in a concentration- and time-dependent manner accompanied by increases both in total cellular cPLA2 activity and in cPLA2 protein levels detected by Western blot analysis. The parallel increase in total cellular cPLA2 activity and cPLA2 protein level indicates that IL-1 beta may induce the synthesis of cPLA2. Incubation of the cells with 10 microM AACOCF3 for 24 h significantly inhibited IL-1 beta-induced PGE2 production strongly suggesting that cPLA2 mediates IL-1 beta-induced PGE2 formation. In unstimulated cells, there is appreciable total cellular cPLA2 activity and protein, but these cells produce low amounts of PGE2 until stimulated by IL-1 beta, suggesting that cPLA2 translocation from cytosol to the membrane is necessary for its bioactivity. In contrast to IL-1 beta, treatment with phorbol ester (12-O tetradecanoyl phorbol-13-acetate, TPA, 10(-10)-10(-6)M) for 24 h significantly inhibited total cellular cPLA2 activity in a concentration-dependent manner. The amount of total cellular cPLA2 protein seen on Western blot remained unchanged following TPA treatment. These data suggest that in WISH cells, IL-1 beta induces both translocation to the membrane and de novo synthesis of cPLA2 protein to sustain prostaglandin (PG) synthesis. In contrast, TPA may only cause cPLA2 translocation but no increase in cPLA2 protein synthesis, resulting in limited PG synthesis. Our results provide a mechanism for the effect of IL-1 beta on prostaglandin synthesis in human amnion cells and provide support for a role of cPLA2 in the mechanism initiating human parturition. PMID- 7480806 TI - PGE2 induces the transition from non-adherent to adherent bone marrow mesenchymal precursor cells via a cAMP/EP2-mediated mechanism. AB - When mesenchymal precursor cells from bone marrow are cultured in the presence of dexamethasone, the existence of distinct non-adherent and adherent populations can be demonstrated. The addition of PGE2, forskolin, or dibutyryl-cAMP can induce a transition from the former to the latter and this may be an important mechanism in the bone anabolic effects of PGE2. On the other hand, phorbol 12 myristate 13-acetate (PMA), an activator of protein kinase C, and sulprostone, an agonist for the PGE2 receptor EP1/EP3 subtypes, had no effect. The phosphodiesterase inhibitor, isobutylmethylxanthine (IBMX), had a synergistic effect in combination with PGE2, whereas neomycin, an inhibitor of inositol phosphate activity, had no effect, and LiC1, an inhibitor of inositol triphosphate metabolism, had an inhibitory effect on the PGE2-induced transition. Consistent with this, the addition of PGE2 to non-adherent bone marrow cells caused a 100% increase in cAMP synthesis. These results suggest that the induction of the transition from non-adherent to adherent osteoblast precursor is mediated by the EP2-PGE2 receptor subtype via an increase in intracellular cAMP synthesis. PMID- 7480808 TI - [Transsexual wish and and cis-sexual defense]. AB - The desire of trans-sexual patients to appropriate the physical attributes of the opposite sex via hormone treatment or surgery confronts psychoanalysis and sex research with problems that are not only psychological in nature. Sigusch uses the term "defence" here not only in a technical sense but also in an epistemological, discourse-analytic and disposition-analytic register. The general dispositions determining the reality of gender and gender difference are "cis-sexual" in nature. The polarity they display extends to the physical and manifests itself in the form of sexual binarism. The author coins the consciously neologistic term "cis-sexual" to describe individuals where physical gender and gender identity are completely and "naturally" co-extensive. Only the realization that "second-nature" is in reality primary can trigger the detotalization of so called trans-sexuality that Sigusch is concerned to call into question. PMID- 7480807 TI - Discordance between macrophage arachidonate metabolic phenotype and the expression of cytosolic phospholipase A2 and cyclooxygenase. AB - Macrophages (M phi s) exhibit variations in their ability to release and metabolize arachidonate (AA) depending on their state of activation, differentiation, and tissue origin. In order to understand these variations on a molecular level, we determined whether differences in AA release and metabolism by murine peritoneal M phi s could be explained in terms of cytosolic phospholipase A2 (cPLA2) and cyclooxygenase (COX) expression. Resident M phi s exhibited greater COX capacity (conversion of exogenous AA to PGE2) but lower phospholipase (PLase) activity (release of endogenous AA) than elicited M phi s. Activation of resident M phi s in vivo with endotoxin increased both their PLase activity and COX capacity. Despite the observed differences in PLase activity, peritoneal M phi s under all conditions expressed similar amounts of cPLA2 mRNA and protein. All M phi s exhibited COX-1 mRNA and protein (i.e., the constitutive isoform of COX), although elicited M phi s exhibited increased mRNA for COX-1 but decreased levels of protein, relative to resident M phi s. Elicited (but not resident) cells also exhibited COX-2 mRNA but not COX-2 protein (i.e., the inducible form of COX). Despite the increased COX capacity of resident cells with in vivo activation, their expression of COX-2 mRNA and protein was equivalent to that of unactivated cells, becoming apparent only after cell adherence in vitro. In sum, there is no simple relationship between the ability of M phi s to release and metabolize AA, and the expression of cPLA2 or COX isoforms. Moreover, adherence appears to be important for the expression of COX-2 by M phi s. PMID- 7480809 TI - [Anna, Irma and Dora--the key to the mothers in the creation of psychoanalysis]. AB - As a science caught up between the mythical and the rational, psychoanalysis is subject to the pulls of two opposing currents, a creative enlightened one concerned with the investigation of the unconscious, and a conservative counter current observable in the psychoanalytic theory of femininity. In her analysis of these warning elements the author goes back to the genesis of psychoanalysis and its creator and takes a close look at the primal myth (the legend of Breuer and Anna O.), the primal dream (Irma), and finally the primal analysis (the case of Dora). King's analysis demonstrates that Freud unconsciously appropriated female psychic features and--in unconscious identification with the mother of the Urszene, bound up with adolescent regression and attendant bi-sexual grandeur fantasies--imagined himself as ideal woman and "conceiver". By appropriating female potency for himself and thus making the opposite sex superfluous, Freud's theory of femininity leaves the place of women vacant, relegating them to the role of a defective entity, a castrated man, rather than acknowledging their otherness. In King's view, this phallic monism is inextricably interwoven with adolescent bisexual identifications and fantasies of grandeur about Freud's own gender, while denying gender-difference genuine recognition and integration. PMID- 7480811 TI - [Troubling femininity. On the topic of adolescence]. AB - Adolescence is of cardinal importance for gender identity, a period of self differentiation in which sexual identity is finally and fully established. The author proceeds from a somato-psychic relation model taking shape between mother and daughter at birth and based on unconscious projections on the part of the mother. The model centres around the contrast between the body as envelope and as (empty) cavity. In the course of adolescence, in which femininity, sexuality and motherhood are supposed to manifest themselves and unite in the female body, girls are faced with the difficult task of relinquishing the idea of their body as a cavity and identifying with the idea of physical containment. With reference to the case of a bulimic adolescent and more general remarks on anorexia, the author traces the difficulties encountered on the arduous path to the appropriation of femininity, which, if strayed from, can lead to the brink of death. PMID- 7480810 TI - [Between idealization and devaluation--problems and perspectives of theoretical analyses of female homo- and heterosexuality]. AB - Female homosexuality remains a controversial topic. Comparing psychoanalytic and sociological-feminist approaches to the phenomenon, the author points up clear theoretical distinctions between the two perspectives. Feminist authors tend towards an idealization of female homosexuality, while psychoanalysts display an inclination to devalue it as something pathological. However, the latent messages in both discourses have a number of points in common, traceable to a key problem they both have to come to grips with, the "lesbian complex". In lesbian relationships Flaake discerns a "third chance", an area not sanctioned by society, in which it is possible to re-attempt something left unattained in the early mother-daughter relationship: the appropriation of the female body. PMID- 7480812 TI - [Difficulties in becoming a woman and staying a woman. On the problems of female identity in menopause]. AB - With a very few exceptions, the sparse psychoanalytic literature on menopause has regarded it almost exclusively as a kind of ailment, a process of decay and loss. By contrast, recent feminist discourse has demonstratively set itself apart from Freud's theory of femininity and emphasized the opportunities that the menopause brings, albeit at the expense of female sexual desire. Hettlage-Varjas and Kurz suggest that critical review of Freud's theories and more recent concepts of femininity can be integrated into psychoanalytical theory on the psychology of conflict and instinctual drives. They thus avoid the equally dangerous temptations either of classifying the female climateric as pathological or else denying the anxieties and sense of loss that involves and declaring it a conflict free zone. PMID- 7480815 TI - Accord VIII--summary. "Sustaining growth and fairness". PMID- 7480816 TI - Issues in occupational health and safety. Gluteraldehyde causes nurse's asthma. PMID- 7480814 TI - [Castration anxiety and oral envy in sexual relations. Analytic studies with ethnological observations]. AB - Ethno-psychoanalysis is the study of the unconscious in foreign cultures. It can however also be of use in understanding unconscious elements operative in our own culture. To illustrate the point, the author describes the "Koro" epidemic which occurs periodically on the island of Hainan off the south of coast of China. This epidemic largely affects young unmarried men and the author pinpoints the unconscious conflicts underlying this collective phenomenon. It transpires that the epidemic is in effect a species of rite of passage in which a group of young males make the communal attempt to overcome castration anxieties which are themselves the product of covert gender envy. In the second section, Gerlach reports on his psychoanalytic encounter with a young man in Germany displaying similar symptoms. The profounder dimensions of this condition were only comprehensible to the author on account of his knowledge of "Koro". PMID- 7480813 TI - [The glass woman]. AB - The author recounts the case of history of a woman patient seeking psychoanalytic treatment for a variety of extremely severe symptoms. In the course of treatment the original symptom constellation changed, revealing new facets but never disappearing completely. Discussing the compulsive, phobic symptomatology of the patient in terms of the traumatic sexual conflicts underlying them and the attendant break with outward reality and psychotic fabrication of a world of the patient's own making, Zeul warns against premature nosological classification. She contends that, in a case like the present one, the diagnosis of the disturbance into a neat set of nosological compartments- borderline/hysteria/psychosis etc.--makes little sense and should be supplanted by an attempt to describe the psychic mechanisms of mental illness. PMID- 7480817 TI - The new Health Minister. PMID- 7480818 TI - Measuring productivity in health care. PMID- 7480819 TI - Epidermoid carcinomas of the floor of mouth treated by exclusive irradiation: statistical study of a series of 207 cases. AB - PURPOSE: Our aim was to study the results and the complications of this series of 207 patients treated from 1976 to 1992 and to compare them with the results of the literature. MATERIAL AND METHODS: The treatment consisted of a combination of external beam irradiation (EBI) (to the tumor and the node areas) and complementary brachytherapy to the primary tumor in 105 cases. In 102 cases, brachytherapy only was delivered to the tumor with or without neck dissection of the node areas. The brachytherapy was performed mostly with the hairpin technique in the beginning (in 61 cases) and subsequently the plastic tube technique with iridium wires. The dosimetry followed the Paris system rules. There were 41% T1, 48% T2, 8% T3, 2% T4 and 1% Tx with 83% N0, 12% N1, 3% N2, 2% N3. Ninety-one percent of patients were male and 9% were female. RESULTS: The local control at 5 years is 97, 72 and 51%, respectively, for T1, T2, T3. The specific survival (without the patients dead of intercurrent disease or second cancer) was 88, 47 and 36%, and the overall survival at 5 years was 71, 42 and 35%. Of note, approximately one third of patients died of intercurrent disease or second cancers (70 patients = 34%). Significant factors that can influence the results are: for local control, the size of the lesion T1/T2/T3 (p < 0.0001); for the locoregional control, the absence of node involvement. The following factors are not significant: age and sex (age significant for survival). The significant treatment factors are the safety margin (p < 0.0003), brachytherapy only on T for T2N0 (p = 0.01). A total duration of treatment of less than 50 days is also significant. The spacing and the total dose (higher dose for large tumors) were not found to be significant. The complications were classified into four grades: grade 1, 17% (median healing, 2 months for soft tissue complications and 5 months for bone complications); grade 2, 12%; grade 3, 6% (frequently requiring surgical resection); grade 4, 0.5% (death, one case). In this series, for the complications, the dose rate, the spacing and the total dose were found not to be significant. CONCLUSION: Carcinomas of the floor of mouth treated by exclusive irradiation have a rather good prognosis and the number of grade 2 and 3 complications remains acceptable. Exclusive brachytherapy is preferable to the combination of EBI plus brachytherapy for T1T2N0. PMID- 7480820 TI - Indications, techniques and results of postoperative brachytherapy in cancer of the oral cavity. AB - PURPOSE/OBJECTIVE: We tried to reduce the number of local recurrences after surgery and external beam irradiation (EBI) in carcinoma of the oral cavity, when margins were positive or close. MATERIAL AND METHODS: From 1980 to 1992, we treated 97 cases of carcinomas of the oral cavity by postoperative brachytherapy. Surgery was combined with EBI+brachytherapy in 51 cases and with brachytherapy alone in 46 cases. We treated 29 T1, 34 T2, 30 T3T4 and four Tx (73% were N0 at first examination and 23% had positive nodes). The type of surgery is analysed. Brachytherapy was performed in one or two planes along the surgical scar. If the mandibular rim was resected, especially when the tongue or the remaining floor were sutured to the internal face of the inferior lip or to the buccal mucosa, the bridge technique was used. To decrease the dose to the inferior part of the mandible, the bridge was modified thanks to experimental dosimetry. RESULTS: At 5 years, the local control (LC) is 89%, the locoregional control (LRC) 82%, the specific survival (SS) 74% and the overall survival (OS) 67%. COMPLICATIONS: We noted 19% of grade 1 (minor), 12% of grade 2 (moderate) and 6% of grade 3 (major) complications. CONCLUSION: Compared with the results of the literature, we think that postoperative brachytherapy can improve classical radiosurgical results in selected cases with a risk of local recurrence. PMID- 7480821 TI - Low dose rate versus high dose rate intraluminal brachytherapy for malignant endobronchial tumors. AB - Although the evolution from low dose rate to high dose rate brachytherapy for malignant endobronchial malignancies was primarily based on economy, patient convenience, and radiation protection, the difference in therapeutic index, if any, between these two modalities must be kept in mind. Our experience with both methods permits assessment of the feasibility of replacing low dose rate brachytherapy with high dose rate brachytherapy. Results with our first 110 patients (group 1) treated with low dose rate brachytherapy (133 procedures) were compared with results with our initial 59 consecutive patients (group 2) treated with high dose rate brachytherapy (161 procedures). In group 1, patients were treated with one or two sessions of 30-60 Gy each calculated at a 1 cm radius. In patients in group 2, we aimed at three weekly sessions of 7 Gy each calculated at a 1 cm radius. External beam irradiation therapy had previously been given to 88% of patients in group 1 and to 85% of patients in group 2. Laser bronchoscopy was performed in 36% of patients in group 1 and in 24% of patients in group 2 before brachytherapy. Clinical or bronchoscopic improvement was noted in 72% of patients in group 1 and in 85% of patients in group 2 (p > 0.05). Complication rates were low and comparable. Survival was similar in both groups (median < 6 months). Although both low dose rate and high dose rate brachytherapy appear equally effective in palliation for malignant endobronchial obstruction, we are now practicing the latter exclusively. PMID- 7480822 TI - Postoperative radiotherapy in carcinoma of the cervix: treatment results and prognostic factors. AB - In order to assess the role of postoperative radiotherapy and prognostic factors, 126 patients who were treated with radiotherapy after surgery for clinical early stage carcinoma of the cervix were reviewed. All patients received external pelvic radiotherapy and 37 patients were treated with additional vaginal cuff irradiation. The 5-year overall survival, disease-free survival and locoregional control rates were 71.1, 69.9 and 78.1%, respectively. The 5-year disease-free survival rates were 40% for grade 3 vs. 75.4% for grade 1 tumours (p = 0.05), 76.5% for pathological stage IB versus 54.1% for pathological stage IIA (p = 0.04), 36.6% for node-positive patients versus 82.5% for node-negative patients (p = 0.0017), 54% for full thickness cervical invasion versus 100% superficial cervical invasion (p = 0.01), 34.8% for positive margins versus 78.1 for negative margins (p < 0.0001). After a multivariate analysis, tumour grade (p = 0.026) and presence of positive margins (p = 0.006) were found to independently influence the outcome. Grade II and III complication rate was 5.5% in all patients. In conclusion, postoperative radiotherapy should be used in patients treated with simple hysterectomy as well as those treated with radical hysterectomy with unfavorable pathological findings. PMID- 7480823 TI - Radiation treatment of cervical lymph node metastases from an unknown primary: an analysis of outcome by treatment volume and other prognostic factors. AB - One hundred and forty-four patients with diagnosis of carcinoma involving cervical lymph nodes from an unknown primary site were seen at the Princess Margaret Hospital, Toronto between 1970 and 1986. This paper addresses the outcome of two treatment approaches using radiation treatment as potentially curative therapy. The treatment approaches compared in a retrospective analysis are radiation to the involved node regions alone (85 cases), and radiation to both the nodes and to potential primary sites in the head and neck (59 cases). The overall 5-year survival rate of the entire group of 144 cases was 41%. There was a trend to increased survival in favour of the group receiving radiation to the nodes and potential primary sites (p = 0.07), however, when differences in extent of nodal involvement were considered and adjusted for using the Cox Proportional Hazard Model, no difference in survival, or cause-specific survival was found between the two treatment groups (p = 0.18 and 0.22, respectively). A total of seven head and neck primary cancers were subsequently discovered, six in the group receiving radiation only to the nodes and one in the group receiving radiation to the nodes and primary sites. The results of this retrospective analysis need to be interpreted with caution but suggest that, for some patients with this diagnosis, radiation to the involved node region alone is adequate. A discussion of the literature is presented with attention to the effect of extent of treatment on outcome. PMID- 7480824 TI - Intrarectal formalin application, an effective treatment for grade III haemorrhagic radiation proctitis. AB - Haemorrhagic radiation proctitis (HRP) is infrequently seen amongst the patients who are either undergoing or have undergone radiotherapy to the pelvis. We treated 16 documented cases of HRP, who did not respond to conventional steroid retention enemas, with 4% formalin application. It was observed that the rectal bleeding was controlled completely in 81% cases in median follow up of 11 months (range 6-17 months) and diversion colostomy could be avoided in all the cases. The effectiveness of local formalin application in severe HRP is described in this article. PMID- 7480826 TI - A mathematical model of the volume effect which postulates cell migration from unirradiated tissues. AB - PURPOSE: In order to simulate the large variation in tolerance doses for very small treatment volumes, we introduce a model which assumes the presence of cells which have migrated from unirradiated tissues. METHODS AND MATERIALS: In order to represent serial architecture, the new model adds a new parameter to the familiar expression for serial architecture. Data derived from the model is fitted to the dose-response data developed by Hopewell et al. (Hopewell, J.W., Morris, A.D. and Dixon-Brown, A. The influence of field size on the late tolerance of the rat spinal cord to single doses of X rays. Br. J. Radiol. 60: 1099-1108, 1987) using white matter necrosis of rat spinal cord. RESULTS: The new model with a cell migration term more accurately describes the large differences in threshold doses for a very small treatment volume than a model without a cell-migration term. CONCLUSION: Although these results do not prove that cell migration is the mechanism behind the volume effect for very small volume, they do suggest that the probability of normal tissue complication is more accurately predicted by the new model. PMID- 7480825 TI - Toxicity, biodistribution and radioprotective capacity of L-homocysteine thiolactone in CNS tissues and tumors in rodents: comparison with prior results with phosphorothioates. AB - L-Homocysteine thiolactone (L-HCTL) was evaluated for its potential as an intravenously-administered central nervous system (CNS) radioprotector in C3H mice and F344 rats. Toxicity assessments in the mouse yielded a LD50 of 297 mg/kg and in the rat 389 mg/kg. Biodistribution studies in tumor-bearing mice showed that brain specimens contained more label at 10 min than the tumors but less at 30 or 60 min. Brain uptake relative to the tumors, the brain/tumor ratio, ranged between 0.5 and 3.3. The cervical spinal cord of non-tumor-bearing rats was irradiated with 32 Gy 137Cs with or without prior treatment with L-HCTL following which the time to forelimb or hindlimb paralysis was measured to determine the relative protective factors (RPFs) for this radiation dose. For forelimb paralysis the RPF was 1.9 (+/- 1.0, SD) and for hindlimb it was 2.0 (+/- 1.1, SD). 36B-10 glioma cells irradiated in vitro with or without L-HCTL and assayed for colony forming capacity demonstrated a dose modifying factor (DMF) of only 1.15 (+/- 0.16, SE). Rats bearing intracerebral 36B-10 glioma received 137Cs irradiation with or without L-HCTL after which the tumors were similarly assayed in vitro. From this the glioma DMF was 1.2 (+/- 0.30, SE). Compared to prior results with phosphorothioates our data show that the toxicity of L-HCTL is roughly the same as WR2721, WR77913 and WR3689 and that it distributes at higher levels in the CNS after systemic administration. L-HCTL may well equal these phosphorothioates at protecting normal CNS tissue without requiring administration directly into the cerebrospinal fluid-containing spaces and it does not protect the 36B-10 glioma. PMID- 7480827 TI - Clinical implementation of an objective computer-aided protocol for intervention in intra-treatment correction using electronic portal imaging. AB - In order to test the feasibility of a protocol for intra-fractional adjustment of the patient position, during radiation therapy treatment in the pelvic region, a two-fold study is carried out. The protocol involves an objective quantitative measurement of the error in positioning starting from the comparison of a portal image with a reference image. The first part of the study applies the protocol to determine the efficacy of adjustment using subjective determination of the positioning errors by a clinician by measuring the residual errors after adjustment. A group of 13 patients was followed extensively throughout their treatment, analyzing 240 fields. In the second part the measurement itself determines the extent of readjustment of the position. Throughout the procedure elapsed time is measured to determine the extra time involved in using this procedure. For this part a group of 21 patients was followed yielding statistics on 218 fields. Using this computer aided protocol it is shown that systematic as well as random errors can be reduced to standard deviations of the order of 1 mm. The price to pay however is additional treatment time up to 58% of the treatment time without the protocol. Time analysis shows that the largest part of the added time is spent on the readjustment of the patients' position adding a mean of 37% of time to the treatment of one field. This is despite the fact that the readjustment was performed using a remote couch controller. Finally a statistical analysis shows that it is possible to select patients benefiting from the use of such a protocol after a limited number of fractions. PMID- 7480828 TI - A diagnostic-quality electronic portal imaging system. AB - Initial clinical experience is presented, on the use of a prototype portal imaging system which is designed to provide diagnostic-quality images on-line at the accelerator. The system comprises a compact diagnostic X-ray unit mounted on the accelerator head, with its source in the isocentric plane exactly 37 degrees around from the therapy source. The image detector is an image intensifier with digital image storage/processing facilities. Images were taken at the accelerator of treatment fields in seven patients (pelvic, head and neck, chest fields) and these images were comparable in anatomical contrast to simulator radiographs. Two techniques for marking the therapy field onto the portal images were successfully demonstrated. One was to mark relevant corners or edges of the therapy light field on the skin using small Pb markers which become clearly visible in the 'diagnostic' portal image. The other was to record a separate 'therapy' image and through the software, extract and superimpose the field edges on to the 'diagnostic' image. The system and method proved fundamentally sound on criteria of image quality, geometric precision of rotation between therapy and imaging conditions, and its potential for development as a practical clinical tool. PMID- 7480829 TI - A comparison of conventional 'forward planning' with inverse planning for 3D conformal radiotherapy of the prostate. AB - A radiotherapy treatment plan optimisation algorithm has been applied to 48 prostate plans and the results compared with those of an experienced human planner. Twelve patients were used in the study, and 3-, 4-, 6- and 8-field plans (with standard coplanar beam angles for each plan type) were optimised by both the human planner and the optimisation algorithm. The human planner 'optimised' the plan by conventional forward planning techniques. The optimisation algorithm was based on fast simulated annealing using a cost-function designed to achieve a homogenous dose in the 'planning-target-volume' and to minimise the integral dose to the organs at risk. 'Importance factors' assigned to different regions of the patient provide a method for controlling the algorithm, and it was found that the same values gave good results for almost all plans. A study of the convergence of the algorithm is presented and optimal convergence parameters are determined. The plans were compared on the basis of both dose statistics and 'normal-tissue complication-probability' (NTCP) and 'tumour-control-probability' (TCP). The results of the comparison study show that the optimisation algorithm yielded results that were at least as good as the human planner for all plan types, and on the whole slightly better. A study of the beam-weights chosen by the optimisation algorithm and the planner revealed differences that increased with the number of beams in the plan. The planner was found to make small perturbations about a conceived optimal beam-weight set. The optimisation algorithm showed much greater showed much greater variation, in response to individual patient geometry, frequently deselecting certain beams altogether from the plan. The algorithm is shown to be a useful tool for radiotherapy treatment planning. For simple (e.g., three-field) plans it was found to consistently achieve slightly higher TCP and lower NTCP values. For more complicated (e.g., eight-field) plans the optimisation also achieved slightly better results with generally less numbers of beams, unfavourable beams being deselected from the plan. Probably the greatest benefit is the reduced time taken by the optimisation to compute optimised beam-weights. This time always < or = 5 min; a factor of up to 20-times faster than the human planner. PMID- 7480830 TI - Intracytoplasmic Sperm Injection. Proceedings of a symposium. Queensland, Australia, 2-4 October 1994. PMID- 7480831 TI - The potential of intracytoplasmic sperm injection (ICSI) to transmit genetic defects causing male infertility. PMID- 7480832 TI - Cell cycle factors in the human oocyte and the intracytoplasmic injection of spermatozoa. PMID- 7480833 TI - Spermatids as male gametes. AB - Intracytoplasmic sperm injection (ICSI) is becoming increasingly popular in human infertility clinics as an efficient method for the treatment of male infertility. It is proposed that spermatids can be used as substitutes for spermatozoa if men are unable to produce sperm in their testes. At least in the hamster and mouse, the nuclei of round spermatids were capable of participating in syngamy when incorporated into homologous mature oocytes either by microsurgical ICSI or electrofusion. Normal mouse offspring were born after after electrofusion of oocytes with round spermatids. When culture in vitro of spermatogonia and spermatocytes is perfected, then spermatids, transforming spermatids and spermatozoa will all be able to be used as male gametes. PMID- 7480834 TI - Comparative intracytoplasmic sperm injection (ICSI) in human and domestic species. AB - The current clinical use of intracytoplasmic sperm injection (ICSI) for the alleviation of male factor infertility has prompted a re-investigation of sperm injection techniques in a number of animal species. This report examines sperm injection of in vitro matured oocytes in the major domestic species and compares the results with the human. Ovine, bovine and porcine oocytes can undergo fertilization and at least limited development without exogenous activation either prior to or subsequent to injection. Porcine is temperature sensitive during fertilization and the early stages of embryo development. The oocytes of all three domestic species, particularly ovine, have a tendency to activate after the injection procedure regardless of the presence or absence of sperm. The implications for early development studies and the practical use of direct sperm injection for domestic species are discussed. PMID- 7480835 TI - Intracytoplasmic sperm injection (ICSI) versus high insemination concentration (HIC) for human conception in vitro. AB - The use of high insemination concentration (HIC) for in vitro fertilization (IVF) was compared with intracytoplasmic sperm injection (ICSI) in cases of male factor infertility. Sibling oocytes (n = 252) from 24 patients were used, 123 for HIC and 129 for ICSI. Although the incidence of fertilization was decreased with HIC (48% v. 61%), this treatment was nevertheless a viable option for many patients, especially when ICSI was not available. However, there was a higher incidence of cytoplasmic fragmentation of embryos after HIC compared with ICSI (36% v. 10%, P = 0.003) and the outcome was significantly affected by the severity of teratozoospermia. Using a cut-off of 5% normal forms, the incidence of fertilization with HIC for the group with < 5% normal forms was 37% compared with 72% for the group with > 5% normal forms; there was also a significant decrease in cleavage rate (P = 0.05) and the number of regular embryos (P = 0.005), and an increase in cytoplasmic fragmentation (P = 0.006) in patients with < 5% normal forms. No distinction was made between cases of teratozoospermia when ICSI was used. The present study confirms the value of HIC as a first line treatment for male infertility, as long as ICSI remains significantly more expensive and concerns on safety are mooted. However, the use of sibling oocytes for ICSI is recommended, especially in cases with < 5% normal sperm morphology. PMID- 7480836 TI - Sperm preparation for intracytoplasmic injection: methods and relationship to fertilization results. AB - Sperm preparation for intracytoplasmic sperm injection (ICSI) is described and the effect of high speed centrifugation during preparation on fertilization rate is evaluated. No significant differences were found in the 2-pronuclear or abnormal fertilization rates between sibling oocytes injected with sperm prepared by swim-up or mini-Percoll combined with high speed centrifugation. The high fertilization rate obtained with both methods indicates that high speed centrifugation is not necessary to prepare sperm for ICSI. Fertilization rates were also compared for sperm obtained from ejaculates, fresh and frozen epididymal aspirates, and testicular biopsies. High fertilization rates were obtained from all groups but they were significantly higher in those oocytes injected with epididymal sperm (78% per oocyte surviving injection). The high fertilization rate with epididymal sperm may reflect sperm quality or may result from the method of sperm preparation for injection. Fertilization after the injection of sperm from which the tail was dislodged during immobilization was compared with that obtained using intact sperm. A significantly lower rate of 2 pronuclear fertilization was found in those oocytes injected with sperm heads only (55%) compared with intact sperm (68%), although cleavage rates between the two groups were similar. The use of hypo-osmotic medium to select potentially live sperm from an immotile sample is also described and fertilization was obtained after the injection of sperm with a structural defect which were selected using this technique. These results indicate that high fertilization rates can be obtained with ejaculated, epididymal and testicular sperm without special treatment.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7480838 TI - Assessment of fertilization failure and abnormal fertilization after intracytoplasmic sperm injection (ICSI). AB - The assessment of fertilization is an important part of intracytoplasmic sperm injection (ICSI) and oocytes are routinely examined about 17 h after injection using Nomarski differential interference contrast optics. However, it is not possible to conclusively determine the aetiology of fertilization anomalies in this manner, so cytological studies were undertaken to determine the causes of failed and abnormal fertilization after ICSI. Oocytes which exhibited no evidence of fertilization, one pronucleus (PN) or 3 PN were fixed in glutaraldehyde, stained with Hoechst 33342 and examined by fluorescence microscopy to identify PN, metaphase chromosomes, sperm heads and polar bodies. A total of 428 unfertilized oocytes were examined from 170 ICSI cycles. Overall, 82% of these unfertilized oocytes were still at metaphase II (non-activated) while the remaining 18% were activated and had 1 PN and two polar bodies. The majority (71%) of the metaphase II oocytes contained a swollen sperm head, which indicates that the spermatozoon was correctly injected but the oocyte did not activate and complete its second meiotic division. The swollen sperm head was located among the metaphase chromosomes in 4.3% of these oocytes, while in some cases (6.6%), the sperm chromosomes had undergone premature chromosome condensation (PCC). Other aetiologies of failed fertilization in these metaphase oocytes were ejection of the spermatozoon from the oocyte (19%) and complete failure of sperm head decondensation (10%). A similar pattern of anomalies was found in 1 PN oocytes, although the ratios were different (swollen sperm head, 51%; ejection of the spermatozoon, 19%; undecondensed sperm head, 30%). Seventy abnormally fertilized oocytes were also examined, of which 63 had 3 PN and a single polar body, indicating that the unextruded second polar body developed into the third PN. In conclusion, the present study demonstrates that the principal cause of fertilization failure after ICSI is failure of oocyte activation and not ejection of the spermatozoon from the oocyte. It is also apparent that further studies are needed to elucidate the mechanisms that control oocyte activation and sperm head decondensation in injected oocytes. PMID- 7480837 TI - Intracytoplasmic sperm injection: instrumentation and injection technique. PMID- 7480839 TI - Development and implementation of intracytoplasmic sperm injection (ICSI). AB - The purpose of this paper is to elucidate the experimental steps that led to the development of intracytoplasmic sperm injection (ICSI) and its application in the human. ICSI has become the most successful micromanipulation procedure for treating male infertility. A total of 355 in vitro fertilization (IVF) cycles utilizing ICSI are described; 180 couples were previously treated in 509 IVF cycles but achieved no fertilization and 175 couples could not be treated by IVF because of extremely poor semen parameters. Of the 3063 metaphase II (M II) oocytes retrieved, 2970 were injected with a survival rate of 93.6%, yielding 1917 bipronuclear zygotes (64.5%). In 148 patients, a foetal heart was evidenced by ultrasound; 11 of these patients miscarried between 7 and 13 weeks of gestation. The ongoing pregnancy rate was 38.6% (137/355) per retrieval and 40.5% (137/338) per embryo replacement. At the time of writing, there were 22 deliveries and one therapeutic abortion for a trisomy 21 chromosomal abnormality. In addition, 66 singleton, 37 twin, 10 triplet and 1 quadruplet pregnancies were ongoing. The concentration of motile spermatozoa in the ejaculate only slightly influenced the fertilization rate (P < 0.001) and the pregnancy outcome (P < 0.01). A preliminary injection procedure utilizing intracytoplasmic injection of isolated sperm heads was performed in 35 M II human oocytes with resultant fertilization and cleavage rates of 74% and 73% respectively. Skills in ICSI were acquired by injecting hamster and unfertilized human oocytes with human sperm. ICSI can be used to successfully treat couples who have failed IVF or who have too few spermatozoa for conventional in vitro insemination.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7480840 TI - Intracytoplasmic sperm injection--clinical results from the reproductive medicine unit, Adelaide. AB - The clinical results of 391 cycles of intracytoplasmic sperm injection (ICSI) performed between June 1993 and July 1994 are presented in this report. A total of 4797 oocytes were collected, of which 3792 were injected. Of these, 2603 (69%) fertilized, with normal and three pronuclear fertilization rates of 65% and 4% respectively. About 6% of the oocytes were destroyed while denuding and during ICSI. There were 373 (95%) embryo transfers from which 119 pregnancies arose, giving pregnancy rates of 32% per transfer and 30% per cycle, and an implantation rate of 15% per embryo. Of the pregnancies, 98 (82%) were ongoing. Supernumerary embryos were frozen in 44% of the cycles and 61 subsequent transfers of 130 frozen-thawed embryos produced 11 pregnancies (18%). Only 47 (12%) patients had less than 50% of their oocytes fertilized (mean 31%) after ICSI, and of these, 8 had no fertilization of 13 eggs. Nevertheless, 37 of these 47 patients had an embryo transfer and 9 achieved a pregnancy with an implantation rate of 14% per embryo. The percent normal sperm morphology weakly correlated with percent fertilization (r2 = 0.027, P < 0.02) but not with the implantation rate (r2 = 0.003, P > 0.05). Fifty-nine patients with only occasional motile sperm in the ejaculate and 23 patients in whom epididymal sperm were aspirated were treated. The fertilization rates (66% and 70% respectively) and pregnancy rates per transfer (32% and 24% respectively) were comparable in these two subgroups. The overall ICSI results were also compared with 515 cycles of routine in vitro fertilization (IVF) which were performed at the same time.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7480841 TI - Factors affecting success with intracytoplasmic sperm injection. AB - In this study, 141 couples underwent 163 cycles of in vitro fertilization (IVF) and embryo transfer in which the eggs were inseminated by intracytoplasmic sperm injection (ICSI). Overall, 41% of the injected eggs were normally fertilized and 81% of the resulting embryos were suitable for cryopreservation (91 embryos) or uterine transfer. From 153 fresh embryo transfers, 45 ongoing or delivered pregnancies (27.6% per cycle) were achieved, and of the 507 embryos transferred, 54 successfully implanted giving an implantation rate per embryo of 10.7%. Five additional pregnancies did not yield a viable fetus or underwent a spontaneous abortion, giving a miscarriage rate of 10% (5/50). Increased maternal age or a prior diagnosis of failed fertilization after conventional IVF had a significantly negative impact on success. Sperm from the testis and epididymis, those retrieved by electro-ejaculation, and completely immotile ejaculated sperm all gave rise to pregnancies. ICSI reinsemination was used with limited success to rescue failed fertilization cycles, although the implantation rate per embryo was poor (5%). ICSI has greatly improved the ability to use IVF for treating couples with a poor fertilization potential. PMID- 7480842 TI - The use of intracytoplasmic sperm injection for the treatment of severe and extreme male infertility. AB - The outcome of treatment by intracytoplasmic sperm injection (ICSI) is described for patients with severe male infertility. In 296 consecutive cycles, a normal fertilization rate of 69% was achieved with 288 cycles (97%) resulting in embryos suitable for transfer. A total of 32 clinical pregnancies were achieved from the transfer of fresh embryos (clinical pregnancy rate of 12% per transfer) and an additional 44 clinical pregnancies were obtained after the transfer of frozen thawed embryos (clinical pregnancy rate of 16% per transfer). Overall, 57 of the 76 pregnancies were ongoing or delivered. An analysis of outcome in 5 male factor subgroups revealed no significant differences in pregnancy and implantation rates between the categories. However, the fertilization rate was significantly lower in patients with oligoasthenoteratozoospermia and significantly higher in those patients for whom epididymal sperm were used for insemination. The treatment of patients with extreme male infertility is also described; normal fertilization and embryo development were obtained using ICSI in patients with mosaic Klinefelter's syndrome, severe sperm autoimmunity, round-headed acrosomeless sperm (globozoospermia), completely immotile sperm selected by hypo-osmotic swelling and sperm isolated from testicular biopsies. Three ongoing pregnancies were obtained from 6 patients for whom testicular sperm were used. These results demonstrate the value of ICSI in the management of severe male infertility, however, the treatment of some types of extreme male infertility using ICSI may be limited. PMID- 7480843 TI - Clinical results from intracytoplasmic sperm injection at monash IVF. AB - The impact of a modification of the intracytoplasmic sperm injection (ICSI) technique on fertilization and pregnancy rates was examined in a retrospective analysis of 171 consecutive ICSI treatment cycles (156 patients). Patients were selected for ICSI on the basis of severe oligoasthenozoospermia (65 patients) or following conventional in vitro fertilization (IVF) with failed or poor fertilization (70 patients). Seven patients in which epididymal or testicular sperm was used, 10 patients with sperm antibodies and 4 patients with retrograde ejaculation or who required electro-ejaculation were also treated with ICSI. In the first 105 cycles (102 patients), single sperm, rendered immotile, were injected into the ooplasm of 979 metaphase II (M II) oocytes using an established technique (Method 1). In the following 66 cycles (513 M II oocytes injected), the ICSI procedure was modified by increased aspiration of the oolemma to ensure the intracytoplasmic deposition of sperm (Method 2). The patient groups did not differ between the two injection procedures. The normal (two pronuclear) fertilization rate increased significantly (P < 0.001) from 34.3% with Method 1 to 73.1% with Method 2, with no difference in the oocyte degeneration rate (4.3% v. 4.5% respectively). The incidence of failed fertilization was significantly (P < 0.01) reduced from 17.1% (18 cycles) to 1.6% (1 cycle) with the change in technique. As a consequence of the increased fertilization rates with Method 2, more embryos were available for assessment and transfer, and a pregnancy rate per oocyte retrieval of 21.2% was obtained for Method 2. Fertilization, embryo transfer and pregnancies were obtained in all patient groups treated with ICSI.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7480844 TI - Clinical intracytoplasmic sperm injection (ICSI) results from Royal North Shore Hospital. AB - The technique of intracytoplasmic sperm injection (ICSI) was first introduced to the Royal North Shore Hospital in April 1993 as part of a controlled study of 100 patient cycles in which sibling oocytes were inseminated by either subzonal insemination (SUZI) or ICSI. This trial showed direct sperm injection to be superior in terms of fertilization. In that study, 58 embryo transfers of 101 ICSI-derived embryos resulted in 10 pregnancies. No miscarriages have occurred and a total of 10 fetal hearts (9.8% per embryo transferred) were detected on ultrasound. There have been 10 deliveries of 10 babies. Since the beginning of 1994, intracytoplasmic injection has been used exclusively for patients requiring micromanipulation to achieve fertilization. There have been 200 patient cycles with 1650 oocytes collected (8.8 oocytes per cycle). Of these oocytes, 1548 were mature (94%) and were subjected to ICSI, and normal fertilization occurred in 874 (56%) of the injected oocytes. The number of oocytes which cleaved and were suitable for fresh transfer or cryopreservation was 818 (94%). There have been 153 fresh embryo transfers of 326 embryos. Twenty-six pregnancies (17% per embryo transfer) have resulted, 22 of which proceeded to ultrasound examination in which 23 fetal hearts were detected (7% per embryo transferred). Three miscarriages have occurred, leaving 19 ongoing pregnancies. There have been 127 cryopreservation procedures involving 492 embryos. To date, there have been 47 embryo thaw cycles, and 93 of the 115 (81%) thawed embryos survived and were transferred. These 47 embryo transfers resulted in 10 pregnancies (21% per embryo transfer), one of which one has miscarried.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7480845 TI - Microfertilization techniques--the Swedish experience. AB - Intracytoplasmic sperm injection (ICSI) has been studied in this animal research programme since 1990. In 1993, the technique was first applied clinically and up to the present time (September 1994), a total of 456 couples have been studied in 538 cycles. The principal indication for the use of ICSI has been severe male sub fertility as judged by a semen analysis. In addition, men with high titres of antisperm antibodies, blockage of the vas deferens and neurological disorders such as spinal cord lesions have been included in the programme. Men with genetic disorders such as cystic fibrosis and acrosome-deficient spermatozoa have also been treated successfully. The overall fertilization rate using ICSI was 59%, which is similar to the conventional in vitro fertilization (IVF) programme in Goteborg, however, the pregnancy rate per embryo transfer (29%) and the ongoing pregnancy rate per transfer (22%) were slightly lower. The total number of pregnancies was 144 with 111 of the pregnancies either ongoing or already delivered. To date, 36 healthy children have been born following 29 deliveries and no major malformations have been diagnosed. Being the first programme in Scandinavia to perform ICSI, this unit has experienced long waiting lists which indicates that severe male sub-fertility will be one of the major groups for treatment with assisted reproductive technologies in the future. PMID- 7480846 TI - Intracytoplasmic sperm injection (ICSI): the Brussels experience. AB - The present report covers the results of a 26-month period in which 1275 consecutive treatment cycles by intracytoplasmic sperm injection (ICSI) were performed in 919 couples. These couples were afflicted with male factor infertility and had had at least one previous failed conventional in vitro fertilization (IVF) treatment cycle. In other couples, the husband had semen parameters incompatible with conventional IVF or suffered from excretory azoospermia which required microsurgical epididymal sperm aspiration or testicular sperm retrieval. Overall, the 2 pronuclear (PN) fertilization rate was 47.7% per retrieved oocyte-cumulus complex and 66.4% per successfully injected metaphase II oocyte. Embryo transfer was performed in 90.8% of started cycles and 362 clinical pregnancies were recorded, giving a clinical pregnancy rate of 28.4% per started cycle or 31.3% per transfer. In addition, updated results on the outcome of pregnancies after microassisted fertilization are presented. As of 30 August 1994, 416 children have been born. Although 16 major congenital malformations have been observed (3.9%), there appears to be no reason for serious concern as regards the occurrence of major congenital anomalies after ICSI. PMID- 7480848 TI - Going international in nursing research. PMID- 7480847 TI - Fertilizing capacity of epididymal and testicular sperm using intracytoplasmic sperm injection (ICSI). AB - For men with uncorrectable obstructive azoospermia, their only hope of fathering a child is microsurgical epididymal sperm aspiration (MESA) combined with in vitro fertilization (IVF). In 1988, proximal epididymal sperm were demonstrated to have better motility than senescent sperm in the distal epididymis, and it was thought that retrieval of motile sperm from the proximal epididymis would yield reliable fertilization and pregnancy rates after conventional IVF. However, the results to date have been poor, and although a minority of patients achieved good fertilization rates with IVF, the vast majority (81%) had consistently poor or no fertilization and the pregnancy rate averaged only 9%. Recently, intracytoplasmic sperm injection (ICSI) has been successfully used to achieve fertilization and pregnancies for patients with extreme oligoasthenozoospermia. ICSI has therefore been applied to cases of obstructive azoospermia and, in this report, 67 MESA-IVF cases are compared with 72 MESA-ICSI cases. The principle that motile sperm from the proximal segments of the epididymis should be used for ICSI was followed, although in the most severe cases in which there was an absence of the epididymis (or absence of sperm in the epididymis), testicular sperm were obtained from macerated testicular biopsies. These sperm only exhibited a weak, twitching motion. In 72 consecutive MESA cases, ICSI resulted in fertilization and normal embryos for transfer in 90% of the cases, with an overall fertilization rate of 46%, a cleavage rate of 68%, and ongoing or delivered pregnancy rates of 46% per transfer and 42% per cycle. The pregnancy and take-home baby rates increased from 9% and 4.5% with IVF to 53% and 42% with ICSI. There were no differences between the results for fresh epididymal, frozen epididymal or testicular sperm, and the number of eggs collected did not affect the outcome. The results were also unaffected by the aetiology of the obstruction such as congenital absence of the vas deferens or failed vasoepididymostomy. The only significant factor which affected the pregnancy rate was female age. It is concluded that although complex mechanisms involving epididymal transport may be beneficial for conventional fertilization of human oocytes (in vivo or in vitro), none of these mechanisms are required for fertilization after ICSI. Given the excellent results with epididymal and testicular sperm, ICSI is obligatory for all future MESA patients. Finally, the use of ICSI with testicular sperm from men with non-obstructive azoospermia is also discussed. PMID- 7480849 TI - The effects of small shifts in body weight on blood flow and interface pressure. AB - The purpose of this study was to (a) determine whether small shifts in body weight change pressure and blood flow under a bony prominence, and (b) identify the magnitude of change in blood flow and pressure under a bony prominence over time. Blood flow and interface pressure in two positions (lateral oblique/supine) with two small shifts in each position (thigh/rib cage) were evaluated over time in 50 nursing home residents. There was a significant decrease in interface pressure under the trochanter through the small shifts intervention, F = 5.36, p < .01. There was a significant decrease in interface pressure, F = 3.90, p < .05, and a significant increase in blood flow, F = 4.85, p < .05, under the sacrum through the small shifts intervention. Blood flow and interface pressure did not change significantly over time. PMID- 7480850 TI - The efficacy of parent training for promoting positive parent-toddler relationships. AB - The effectiveness of a parent training program for promoting positive parent child relationships was examined among families of 2-year-olds. Forty-six mothers and fathers and their toddlers were assigned to either an intervention or comparison group. Intervention group parents participated in a 10-week program that focused on principles for effectively interacting with their toddlers. Parents completed measures of parenting self-efficacy, depression, stress, and perceptions of their toddler's behaviors and were videotaped playing with their toddlers preintervention, postintervention, and 3 months following the intervention. Repeated measures ANOVAs showed that the parent training program led to significant increases in maternal self-efficacy, decreases in maternal stress, and improvements in the quality of mother-toddler interactions. No significant effects were found among fathers. Explanations for obtaining different outcomes for mothers and fathers are discussed and directions for future research are recommended. PMID- 7480852 TI - Adjustment of women and their husbands to recurrent breast cancer. AB - The psychosocial adjustment of women with recurrent breast cancer (N = 81) and their husbands (N = 74) were compared to determine if they report different levels of adjustment, support, symptom distress, hopelessness, and uncertainty. Women with recurrent breast cancer reported more emotional distress than their husbands, but both had a similar number of psychosocial role problems. Women and husbands differed in the amount of support and uncertainty they reported but not in the levels of symptom distress or hopelessness they perceived. Women, in contrast to their husbands, expressed more surprise that their cancer recurred and found the recurrent phase of illness more distressing than the initial diagnosis. PMID- 7480853 TI - Influence of symptoms, lung function, mood, and social support on level of functioning of patients with COPD. AB - To explain how symptoms, lung function, mood, and social support affect level of functioning, patients (N = 143) with chronic obstructive pulmonary disease (COPD) completed measures assessing their symptoms (Bronchitis-Emphysema Symptom Checklist), mood (Profile of Mood States), social support (Personal Resource Questionnaire), and functioning (Sickness Impact Profile). Those who were receiving oxygen therapy (n = 52) had significantly lower FEV1 scores and experienced significantly poorer functioning than those who were not receiving oxygen therapy (n = 91). Results of path analyses indicated that symptoms and mood directly, and social support indirectly, influenced the functioning of those who were not receiving oxygen. For those who were receiving oxygen, only symptoms directly, and FEV1 indirectly, influenced their functioning. These models need to be confirmed using other samples of patients with COPD. PMID- 7480851 TI - Consistency between maternal employment attitudes and employment status. AB - The stability of mothers' attitudes toward employment, employment status, and consistency between employment attitudes and behavior over a 15-month period was examined. Factors such as family finances, availability of child care, and child's health status that could influence either employment attitudes or employment behavior were explored. Mothers of preterm infants (N = 98) and mothers of full-term, healthy infants (N = 101) completed questionnaires regarding home/employment orientation, choice and satisfaction with the employment decision, and employment behavior as part of a battery of instruments administered when their infants were 3, 9, and 18 months old. Analyses indicated considerable stability in attitudes, behaviors, and consistency over a 15-month period for mothers of both full-term and preterm infants. Consistency groups were created based on employment status (employed versus not employed) and consistency status (consistent versus inconsistent) at each time point. Consistency groups differed on ratings of financial need and availability of child care. PMID- 7480855 TI - The effect of chronic obstructive pulmonary disease on quality of life. AB - Lazarus and Folkman's (1984) theory of stress, coping, and adaptation served as the framework for a path model of quality of life in chronic obstructive pulmonary disease (COPD). It was hypothesized that psychosocial variables (depression, anxiety, self-esteem, optimism, and social support) would mediate the effects of demographic (age, socioeconomic status) and disease (dyspnea, disease severity, and functional status) variables on life quality in COPD. Interview, pulmonary function, and 6-minute walk test data obtained from 126 subjects with COPD resulted in support for the majority of the hypothesized relationships. Variables having direct effects on life quality were self-esteem, depression, social support, and age. Anxiety and optimism did not have their hypothesized effects on quality of life, suggesting the need to reconsider their importance in influencing life quality in COPD. PMID- 7480856 TI - Matching versus cultural competence in research: methodological considerations. AB - Researcher/participant matching has been used in nursing research as a strategy for the development of culturally competent knowledge. In this manuscript, the complexities inherent in matching in nursing research are discussed. In raising and discussing issues and questions related to matching, we argue that although matching may be one of the strategies used to generate culturally competent knowledge, it is frequently too complex for operationalization. Therefore, we propose that researchers reflect on questions related to their knowledge of culture, their cultural sensitivity, and the nature of collaboration during every phase of the research. Reflecting on and responding to these questions in every research project may facilitate the development of more culturally competent knowledge. PMID- 7480854 TI - Psychophysiologic factors contributing to functional performance in people with COPD: are there gender differences? AB - The purpose of this study was to compare the functional performance profiles of men and women with chronic obstructive pulmonary disease, describe the extent to which physiologic impairment, physical symptoms, and psychosocial resources contribute in a cumulative manner to performance, and outline the extent to which these contributions differ across gender. Secondary data analyses were employed. Although women (n = 45) reported more functional difficulty than men (n = 44) in 9 of 12 Sickness Impact Profile categories, the differences were not significant. Using hierarchical regression procedures, physiologic, symptomatic, and interactive variables predicted total (R2 = .64) and physical performance (R2 = .52), while symptomatic and psychosocial variables predicted psychosocial performance (R2 = .53). Results indicate that models of functional performance may be different for men and women. PMID- 7480859 TI - . . . and even more on TT. PMID- 7480857 TI - Triangles and crystals: on the geometry of qualitative research. AB - Triangulation has become increasingly appealing to researchers in nursing as a device to grasp the complexity of human phenomena, operationalize the holistic elan of nursing, and to accommodate both qualitative and quantitative approaches to inquiry. Yet, a misplaced ecumenicism, definitional drift, and conceptual misappropriation are evident in discussions of triangulation, which has become a technique for everything. Moreover, the triangle is somewhat lacking as image and metaphor for qualitative inquiry. As an idea and technique (in)formed by the triangle, triangulation should be used only to refer to a distinctive strategy for confirmation employed within research paradigms in which convergent and consensual validity are valued, and in which it is deemed appropriate to use information from one source to corroborate another. PMID- 7480858 TI - . . . and even more on TT. PMID- 7480860 TI - Re: Replication research in nursing. PMID- 7480861 TI - Colorectal cancer. Epidemiology, etiology, pathogenesis and prevention. AB - Colorectal cancer (CRC) is a neoplasm with a steadily growing incidence in Western countries. Moreover the age of 50 and over is a critical risk factor. The relationship between dietary, environmental factors and CRC has been evaluated. At present, a number of genetic risk factors such as the genetic susceptibility, tumor suppressor genes and oncogenes are thought to play a major role. The correlation of CRC with adenomatous polyps and chronic inflammatory disease, ulcerative colitis in particular, has been evaluated. Prevention of CRC is mainly based on a rational diet and in the adoption of mass screening programmes. PMID- 7480862 TI - Radiotherapy in rectal cancer: stage of the disease and therapeutic approach. AB - Rectal cancer requires treatments tailored according to the stage of the disease at diagnosis. In stage I lesions exclusive surgery is the elective treatment. Radiotherapy is used in those cases where the site of the lesion would sacrifice sphincter function. When the features of neoplasm are favourable (size < 5 cm, grading 1-2, histological type adenocarcinoma) local excision and postoperative radiotherapy enable to achieve results comparable to those of radical surgery. In operable lesions (T3-T4 (vagina) N0-2, M0) at high risk for local recurrence, high dose preoperative radiotherapy in some randomized studies has shown a significant improvement in local control and survival with a low toxicity, especially in patients undergoing radical surgery. Concomitant chemotherapy in postoperative radiotherapy has significantly improved local control and survival as compared to radiotherapy alone. Continuous infusion appears to reduce the toxicity, which is otherwise high. Randomized studies of preoperative radiochemotherapy vs radiation alone or vs postoperative radiochemotherapy are in progress in various Centers. Intensification with radiotherapy alone (IORT, fractionation) is also under study. In lesions unresectable for cure (T4, N0-3, M0) radiotherapy alone or combined with surgery did not show significant results. Intensification with concomitant chemotherapy and/or with IORT has enabled over 70% local control and 50% 5-year survival. PMID- 7480863 TI - Combined modality staging of low risk rectal cancer. AB - The work-up conducted in order to reach a correct diagnosis based on a combined modality staging in patients with low risk rectal cancer, is presented. The diagnostic accuracy of each imaging technique is analyzed by the comparison of the radiologic TN with the histologic TN in 13 patients directly referred to surgery based on combined modality staging. Control with histologic findings has staged as T1-T2 13 patients directly referred to surgery, while only a patient was shown to have microinfiltration of perirectal fat. As for T, diagnostic accuracy was 90% for transrectal US, 66% for MRI, 61% for CT. Accuracy of transrectal sonography in the differentiation of T1 from T2 was 70% with a trend towards overstaging. None of the procedures has suggested the presence of metastatic lymph nodes while on histology small neoplastic foci were evidenced in a normal-sized perirectal lymph node. The high diagnostic accuracy of US in the definition of T in rectal cancer is confirmed. It is underlined that no procedure enables the detection of neoplastic microinfiltration of normal lymph nodes. PMID- 7480865 TI - Combined modality staging of high risk rectal cancer. AB - The personal experience with the radiologic staging of high risk rectal cancer undergoing preoperative treatment is reported. 61 patients shown to be affected by locally advanced (T3-T4, N+with any T) rectal tumors on combined modality staging, underwent restaging 4-5 weeks after treatment. In all patients liver US, pelvic CT and colorectal enema were performed before and after treatment. In 22 patients with low rectal cancer transrectal sonography was performed. In 19 patients pelvic MRI was performed. The 61 patients were considered operable on second staging and thus referred to surgery. Subsequent control on histology has confirmed the diagnostic accuracy of the single procedures and of their combination. US was shown to be of high accuracy in the evaluation of T (90.8%) and of high predictive value for N. CT accuracy (84%) and MRI accuracy (78.9%) was lower because both tend to overstaging in the evaluation of T of rectal tumors undergoing preoperative treatment. Low CT accuracy (64%) and MRI accuracy (58%) was observed for N. There was optimum agreement between histology and imaging in the assessment of tumor shrinkage, well visualized by rectal enema which supported combined restaging with 88% accuracy for T. PMID- 7480864 TI - Combined modality therapy in low risk (T2N0) rectal cancer. AB - The authors' experience with local excision (LE) and adjuvant radiotherapy in the treatment of selected cases of rectal cancer, is reported. 41 patients with distal rectal cancer underwent elective LE for cure. Selection criteria were: the site of tumor in the lower rectum, exophytic growth, maximum diameter equal to or lower than 4 cm, tumor "freely" mobile on the rectal wall, clinical staging T1-2 N0M0, histological grading G1-2. Patients shown to be T2 on definitive histology underwent adjuvant radiotherapy to the site of tumor and to pelvic lymph nodes. LE was performed via transanal route under general anesthesia. Operative mortality was 0% and morbidity 7.3%. In 37 cases (90%) surgery was considered radical and curative. The incidence of local recurrence was 5.4%, overall evidence of disease 8.1%, cancer-specific mortality 5.4% and 5-year actuarial survival 90%. The combination with radiotherapy has achieved similar results in T1 (22 cases) and T2 (15 cases) tumors. It is concluded that LE combined with radiotherapy in T2 tumors in selected cases represent a valid therapeutic alternative to more demolitive surgery. PMID- 7480866 TI - Combined modality therapy of resectable high risk rectal cancer. AB - Aim of this phase I-II study was to evaluate the efficacy of preoperative concomitant radiochemotherapy in resectable high risk (TNM stage: II and III) rectal tumors, 64 patients entered the study: 37 had low rectal cancer, 27 mid rectal cancer. 50 patients were clinically staged as stage III (Dukes C) and 14 as stage II (Dukes B). Treatment protocol included bolus mitomycin C at the dose of 10 mg/m2 on day 1 and 5FU continuous infusion at the daily dose of 1000 mg/m2 on day 1, 2, 3, 4. Concomitant external radiotherapy up to a dose of 3780 cGy was delivered at the daily dose of 180 cGy. Surgery was performed 4 to 5 weeks after radiation therapy (RT). Before surgery all patients were clinically restaged to evaluate the response to concomitant radiochemotherapy. Treatment compliance was 97%. Toxicity was 27% prevalently shown as bone marrow depletion and radiodermatitis. In 37 patients (61%) there was 50% reduction (partial response) of neoplastic volume. In 5 patients (8%) no neoplastic cells were evidenced in the surgical specimen on histology (complete response). The distance between the lower margin of the tumor and the internal anal orifice increased in 72% of cases. Postoperative morbidity was 28%. The incidence of anastomotic dehiscences was 8.7% over 46 anterior resections. Postoperative mortality was nil. Definitive staging evidenced 24 patients (39%) stage I or with no evidence of tumor. The incidence of local recurrence was 5% and that of distant metastasis 8%. PMID- 7480867 TI - Preoperative radiotherapy and IORT in resectable rectal cancer at high risk for local recurrence. AB - Over a 4-year period 35 patients with mid- (15 patients) and low (20 patients) rectal cancer clinically staged as T2 N1-2, T3 N0-2 underwent a protocol of combined surgery and radiotherapy. The protocol included: preoperative external beam radiotherapy (38 Gy, ICRU50); surgical resection; IORT on tumor bed (10 Gy). Toxicity of preoperative treatment was mild with a single case (2.9%) of grade 3 gastrointestinal toxicity. 18 patients underwent anterior resection and 17 abdominoperineal resection. Perioperative mortality and morbidity were 0% and 17.1% respectively. At a mean follow-up of 25 months all patients were alive. The single anastomotic recurrence observed was rescued with abdominoperineal resection. PMID- 7480868 TI - Preoperative concomitant radiochemotherapy and IORT in locally advanced rectal cancer. AB - 34 patients with locally advanced (T4) or recurrent rectal cancer have been treated with: 1. external beam radiotherapy (45-48 Gy) + 5FU(1000 mg/m2/daily iv continuous infusion day 1-4) + MMC (10 mg/m2/daily iv, day 1); 2. surgical resection (if feasible) + IORT (10-15 Gy); 3. adjuvant chemotherapy (5FU+leucovorin, 6-8 cycles). Grade 3 acute hematological toxicity was observed in 1 case only. 23 of 34 patients underwent radical surgery. Perioperative mortality and morbidity was 0% and 17% respectively. In the 23 operated patients with a mean follow-up of 18.6 months there were 2 local recurrences, 5 blood metastases, (1 death of disease). 16 patients were shown to be NED (3-36 months). PMID- 7480869 TI - Combined modality follow-up and postoperative recurrence in rectal cancer. AB - Rectal cancer patients are at high risk for disease progression even after radical surgery for cure. Prognosis depends on a timely diagnosis for effective, curative therapies. In resected patients, an accurate follow-up especially within the first two years of surgery, is required. Clinical and radiologic follow-up is based on periodical, timed physical and instrumental exams (rectal exploration, neoplastic markers, rectoscopy, colonic enema, CT MRI, TRUS) which supply information on locoregional (liver, lymph nodes, peritoneum, chest) recurrence. Rectal exploration, endoscopy, colonic enema and TRUS enable the control of anastomosis and the identification of possible metachronous tumors. CT and MRI as liver US and chest X-ray, detect recurrence in other sites (pelvic perianastomotic recurrence, peritoneal carcinomatosis, lymph node or liver locations). A diagnostic problem is represented by the difficult differentiation of local recurrence from postoperative fibrosis. In this case, surveillance in the course of time with exams more reliable in tissue differentiation (MRI, though with some limitations) and US- or CT-guided biopsy, is required. PMID- 7480870 TI - The identification of two myoinhibitory peptides, with sequence similarities to the galanins, isolated from the ventral nerve cord of Manduca sexta. AB - Two new myoinhibitory peptides, Mas-MIP I and Mas-MIP II, were identified from the ventral nerve cord of the adult tobacco hornworm, Manduca sexta. Sequences obtained by a combination of automated Edman degradation and electrospray mass spectrometry were, respectively, AWQDLNSAW and GWQDLNSAW. The native peptides were found to co-elute with synthetic C-terminal amides on a reverse phase HPLC system. When applied to isolated ilea (anterior hindgut) of adult M. sexta, both peptides were found to significantly reduce the rate of peristalsis, or abolish peristalsis entirely, at concentrations of 1 x 10(-9) M. Both peptides share sequence similarities with Lom-MIP, a previously identified myoinhibitory peptide from Locusta migratoria, and with the N-terminal portion of vertebrate peptides in the galanin family. PMID- 7480873 TI - Isolation, identification and functional significance of [Hyp2]Met-callatostatin and des Gly-Pro Met-callatostatin, two further post-translational modifications of the blowfly neuropeptide Met-callatostatin. AB - Two post-translationally modified neuropeptides of the Met-callatostatin (Gly-Pro Pro-Tyr-Asp-Phe-Gly-Met-NH2) family have been identified from head extracts of the blowfly Calliphora vomitoria. They are the octapeptide, [Hyp2]Met callatostatin, (Gly-Hyp-Pro-Tyr-Asp-Phe-Gly-Met-NH2) and the truncated hexapeptide, des Gly-Pro Met-callatostatin (Pro-Tyr-Asp-Phe-Gly-Met-NH2). The existence of the [Hyp2]Met-callatostatin variant, in addition to the previously identified [Hyp3]Met-callatostatin peptide, suggests that the motif for prolyl hydroxylation in C. vomitoria is more variable than those known from mammalian and other invertebrate studies where, in those regulatory peptides containing a pair of adjacent prolyl residues so far studied, e.g., bradykinin, and the mosquito peptide Aea HP-I, only one of the pair (the second) is known to undergo hydroxylation. The truncated hexapeptide, des Gly-Pro Met-callatostatin could be produced as a result of the action of a dipeptidyl peptidase II type of enzyme which is known from mammalian studies to be unique in its ability to cleave between the two prolyl residues of an Xaa-Pro-Pro- sequence, where Xaa is any unprotected NH2-terminal amino acid. This enzyme is, however, considered unlikely to be able to cleave the Gly-Hyp-Pro-sequence, which would suggest a functional significance for such a post-translational modification. For this reason, it is of interest that [Hyp2]Met-callatostatin (and earlier, [Hyp3]Met-callatostatin) have been shown to be potent inhibitors of the spontaneous contractions of the hindgut of C. vomitoria (biphasic dose-response curve with IC50 values of 10(-14) M and 10(-7) M).(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7480872 TI - Identification of two allatostatins from the cricket, Gryllus bimaculatus de Geer (Ensifera, Gryllidae): additional members of a family of neuropeptides inhibiting juvenile hormone biosynthesis. AB - Two peptide inhibitors of juvenile hormone biosynthesis, designated G. bimaculatus allatostatins A1 and A2, have been purified from extracts of the brain of the field cricket Gryllus bimaculatus. The primary structures of these peptides were assigned as Ala-Gln-His-Gln-Tyr-Ser-Phe-Gly-Leu-NH2 (Grb-AST A1) and Ala-Gly-Gly-Arg-Gln-Tyr-Gly-Phe-Gly-Leu-NH2 (Grb-AST A2). Each of the peptides shows C-terminal amino acid sequence similarity to cockroach allatostatins and blowfly callatostatins. The two peptides are potent inhibitors of in vitro juvenile hormone production by corpora allata from virgin females of G. bimaculatus. PMID- 7480874 TI - Isolation and primary structure of a novel adipokinetic peptide from the pyrgomorphid grasshopper, Phymateus leprosus. AB - Using a heterologous bioassay, monitoring lipid increase in the haemolymph of migratory locusts, two peptides have been purified from methanolic extracts of corpora cardiaca of the pyrgomorphid grasshopper, Phymateus leprosus. The structures of both peptides were elucidated by a combination of Edman degradation, after deblocking the N-terminal pyroglutamic acid residue, and mass spectrometric techniques. One peptide is an octapeptide (pGlu-Leu-Asn-Phe-Ser-Thr Gly-Trp-NH2) which also occurs in desert locusts; the second peptide is a novel decapeptide member of the AKH-family (pGlu-Leu-Thr-Phe-Thr-Pro-Asn-Trp-Gly-Ser NH2). PMID- 7480871 TI - Isolation and identification of Lom-SG-SASP, a salivation stimulating peptide from the salivary glands of Locusta migratoria. AB - From a methanolic extract of about 2500 salivary glands of Locusta migratoria a peptide was isolated which stimulates cAMP production in the salivary glands and salivation. Maldi-TOFMS revealed a mass of 1779 Da. The primary structure of the peptide is NH2-EVGDLFKEWLQGNMN-COOH. The peptide is named Locusta migratoria Salivary Gland-Salivation Stimulating Peptide (Lom-SG-SASP) because of its simulating effect on salivation. Lom-SG-SASP displays no relevant sequence similarities with any other known peptide from vertebrate or invertebrate sources. The effect of synthetic Lom-SG-SASP on cAMP production in the salivary glands and on salivation is discussed. PMID- 7480875 TI - A novel low molecular weight ecdysiotropin in post-diapause, pre-hatch eggs of the gypsy moth, Lymantria dispar L. (Lepidoptera: Lymantriidae). AB - Extracts of post-diapause, pre-hatch eggs of the gypsy moth, Lymantria dispar L. were examined for prothoracicotropic hormone (PTTH)-like activity using an in vitro assay involving last-instar prothoracic glands (PGs). The eggs were extracted in water, eluted from a low-pressure C18-silica cartridge in 60% acetonitrile, and fractionated on a high-performance, size-exclusion column. The primary ecdysiotropic activity eluted with an estimated molecular weight of 2.1 kDa far below the 4-7 kDa size determined for the low molecular weight PTTHs (bombyxins). Dose-response analysis revealed that the maximum activation was reduced by 75% by organic solvent extraction, but the remaining activity retained the ability to maximally activate the PGs 10-fold in vitro. At least some of the ecdysiotropic activity in the post-diapause, pre-hatch egg is localized in the brain of the pharate larva, and this activity increases dramatically prior to hatch when eggs are incubated at 25 degrees C. PMID- 7480876 TI - An ecdysiostatin from flies. AB - A bioassay for compounds with ecdysiostatic activity ('ecdysiostatins') was developed in order to prove the existence of an ecdysiostatin in blowfly larvae. The factor eluted by HPLC like Neb-TMOF (trypsin modulating oostatic factor), a hexapeptide that inhibits ecdysone biosynthesis. The ecdysiostatic activity of Neb-TMOF is specific, related peptides were less active or inactive. TMOF inhibits ecdysone biosynthesis rather than its secretion. It increases the cAMP level of ring glands. Effectors that elevate the intracellular cAMP can mimic the inhibitory effect of the ecdysiostatin. This suggests that cAMP may control steroidogenesis in fly larvae by shutting ecdysone biosynthesis. Thus, the hexapeptide acts as a prothoracicostatic hormone (PTSH) that controls ecdysone biosynthesis. PMID- 7480877 TI - Feeding the mosquito Aedes aegypti with TMOF and its analogs; effect on trypsin biosynthesis and egg development. AB - Female Aedes aegypti that were given a blood meal by enema deposited yolk in their oocytes and synthesized trypsinlike enzymes in their midgut. When females were given an enema of Aea-TMOF (Trypsin Modulating Oostatic Factor) (NH2 YDPAPPPPPP-COOH) and blood both egg development and trypsin biosynthesis were inhibited. Similar results were observed if TMOF was mixed with the blood meal and fed to female mosquitoes through a membrane. Renin inhibitor (NH2-PHPFHFFVYK COOH) or poly proline given by enema with the blood meal did not affect egg development or trypsin biosynthesis. Feeding of TMOF analogs P1 (NH2-YDPAP-COOH) or P4 (NH2-YDPAPPPP-COOH) inhibited trypsin biosynthesis in the midgut. Injecting or giving an enema of an amidated peptide (NH2-WRPGPPPPPP-CONH2) of HIV-2 X-ORF protein also inhibited egg development and trypsin biosynthesis in the mosquito gut. When [3H]TMOF was purified by high performance liquid chromatography (HPLC) and fed with the blood meal through a membrane to female mosquitoes, [3H]TMOF outside the gut increased linearly for the first 24 h and 28% of the hormone was found outside the gut at 72 h. These results suggest that TMOF and its active analogs traverse the gut epithelial cells into the hemolymph, bind TMOF gut receptor(s) and modulate trypsin biosynthesis. PMID- 7480878 TI - Synergism between diuretic peptides controlling ion and fluid transport in insect malpighian tubules. AB - This study provides clear evidence for synergism between diuretic peptides from Locusta migratoria, Locusta-DP (a CRF-related peptide) and locustakinin (an insect myokinin). At low concentrations, the peptides act co-operatively to stimulate Malpighian tubule fluid secretion by more than the sum of their separate responses. They work via different second messengers, and their synergistic actions can be mimicked with exogenous 8-bromo-cyclic AMP replacing Locusta-DP, and/or with thapsigargin substituting for locustakinin. Additionally, 5-HT, which had been shown previously to stimulate tubule secretion via a cyclic AMP-independent mechanism, potentiated the response to Locusta-DP. Potassium is invariably the dominant cation secreted by locust tubules, but Locusta-DP, whether in the absence or presence of locustakinin, increases JNa at the expense of JK. In contrast, JNa and JK increase in parallel with JV after stimulation by locustakinin. These findings are consistent with the known actions of cyclic AMP and leucokinin-VIII on mosquito tubules. The ability to vary the relative amounts of Na+ and K+ delivered to the hindgut may be important for the maintenance of haemolymph volume and composition in recently fed locusts. PMID- 7480879 TI - Evidence that locustatachykinin I is involved in release of adipokinetic hormone from locust corpora cardiaca. AB - The glandular cells of the corpus cardiacum of the locust Locusta migratoria, known to synthesize and release adipokinetic hormones (AKH), are contacted by axons immunoreactive to an antiserum raised against the locust neuropeptide locustatachykinin I (LomTK I). Electron-microscopical immunocytochemistry reveals LomTK immunoreactive axon terminals, containing granular vesicles, in close contact with the glandular cells cells. Release of AKH I from isolated corpora cardiaca of the locust has been monitored in an in vitro system where the amount of AKH I released into the incubation saline is determined by reversed phase high performance liquid chromatography with fluorometric detection. We could show that LomTK I induces release of AKH from corpora cardiaca in a dose-dependent manner when tested in a range of 10-200 microM. This is thus the first clear demonstration of a substance inducing release of AKH, correlated with the presence of the substance in fibers innervating the AKH-synthesizing glandular cells, in the insect corpora cardiaca. PMID- 7480880 TI - Morphological changes of BM-N4 cells induced by bombyxin, an insulin-related peptide of Bombyx mori. AB - Bombyxin is a brain secretory peptide of the silkmoth Bombyx mori whose amino acid sequence shows considerable sequence homology with vertebrate insulin-family peptides. We found that a Bombyx cell line, named BM-N4, showed morphological changes when bombyxin was added to the culture medium at doses as low as 10(-10) M. Bombyxin-treated cells displayed a series of morphological modifications: 1 to 2 weeks after introduction of bombyxin, the cells increased in size, then they trended to aggregate, or took a spindle shape. These changes of the cells were bombyxin specific, not induced by other vertebrate insulin-family peptides. To prove that these modifications of the cells are mediated by receptors on the cell surface of the cells, we performed a receptor binding assay using 125I-labeled bombyxin. Scatchard analysis of the binding assay indicated that this cell line has a single class of receptors for bombyxin with a Kd = 2.36 +/- 0.56 nM and each cell has 15,800 +/- 1400 binding sites on the cell surface. PMID- 7480881 TI - Intracellular signal transduction of PBAN action in lepidopteran insects: inhibition of sex pheromone production by compactin, an HMG CoA reductase inhibitor. AB - Pheromone biosynthesis activating neuropeptide (PBAN) regulates sex pheromone production in the pheromone glands of many species of female moths. In order to probe the biochemical steps as well as underlying mechanisms regulated by PBAN, we have tested the effect of chemicals on sex pheromone production by using an in vitro assay. Among the chemicals we tested here, compactin, a specific 3-hydroxy 3-methylglutaryl coenzyme A (HMG CoA) reductase inhibitor, clearly inhibited the pheromone biosynthesis in the silkworm, Bombyx mori, and the common cutworm, Spodoptera litura. Since the activation of HMG CoA reductase occurs by dephosphorylation mediated by a specific phosphatase and the biochemical step regulated by PBAN in bombykol biosynthesis is similar to the one catalyzed by HMG CoA reductase in cholesterol biosynthesis, the present results support the idea that phosphoprotein phosphatase has a significant role to regulate bombykol production in the intracellular transduction of PBAN action in B. mori. PMID- 7480882 TI - Characterization of the adipokinetic hormone receptor form the fat body of Manduca sexta. AB - A tritium labeled Manduca sexta adipokinetic hormone (M-AKH) was synthesized (pE L-T-[p3H]F-T-S-S-W-G-NH2) (specific activity 27 Ci/mmol) which was fully active in a bioassay. It was used in a filtration based binding assay to characterize the M-AKH receptor from the fat body of M. sexta. Membrane fractions were prepared from fat body and optimal binding conditions were determined. A Kd of 7.10(-10) M was determined and the receptor concentration estimated to be 0.5 pmol/mg membrane protein. No receptor binding was found when membranes were prepared from brain, heart or flight muscle of M. sexta or from fat body of the cockroach Blaberus discoidalis. However, specific binding was found with membrane preparations from the pterothoracic ganglion of M. sexta. The membranes from the ganglion had a much smaller number of binding sites than the fat body membranes, however, the binding was specific and observed in each experiment. PMID- 7480883 TI - Binding affinity and physiological activity of some HVFLRFamide analogues on the oviducts of the locust, Locusta migratoria. AB - SchistoFLRFamide (PDVDHVFLRFamide) is an insect neuropeptide which inhibits spontaneous and induced contractions of locust oviduct. The active core for inhibition lies within the sequence HVFLRFamide, whereas the core for binding lies within the sequence VFLRFamide. This latter peptide shows activity reversal, possessing minor stimulatory activity. The bioassay and receptor binding assay were employed to define the relationship between the binding affinity and biological activity of HVFLRFamide analogues on the oviduct of Locusta migratoria. Each amino acid in the sequence VFLRFamide was substituted with a structurally similar or dissimilar amino acid to yield a group of HVFLRFamide analogues. These analogues were tested for their binding affinity to receptors in locust oviduct membrane and for their biological effects on contractions of the isolated locust oviduct. The results indicate that (1) with the His residue in position 1, no activity reversal is achieved, the analogues are either inhibitory or possess no biological activity; (2) the C-terminal RFamide group is critical for binding affinity and biological activity of the analogues and (3) substitution of Arg5 or Phe6 with structurally similar amino acids Lys5 or Tyr6 results in two high-affinity antagonists, while substitution of Val2 with Leu2 or Ala2 results in high-affinity agonists. PMID- 7480884 TI - Identification and partial characterization of receptors for allatostatins in brain and corpora allata of the cockroach Diploptera punctata using a binding assay and photoaffinity labeling. AB - We have developed both an in vitro binding assay and a photoaffinity labeling assay to demonstrate and partially characterize putative receptors for allatostatins in brain and in corpora allata of Diploptera punctata. Isolated brain membranes were photoaffinity labeled with 125I-RYBPA (photoaffinity analogue of dip-allatostatin 5). Following labeling with 125I-RYBPA, SDS-PAGE and autoradiography revealed the presence of a putative receptor (37 kDa) for dip allatostatin 5 and dip-allatostatin 7. Specific labeling was demonstrated by dose dependent competition with either dip-allatostatin 5 or dip-allatostatin 7. The in vitro binding assay indicated that the receptor for dip-allatostatin 5 had a Kd of (9.0 +/- 0.9).10(-10) M and Bmax of 2.2 +/- 0.3 pmol/mg membrane protein. For dip-allatostatin 7, two Kd values of (1.5 +/- 0.1).10(-9) M and (3.8 +/- 0.3).10(-9) M were obtained, with Bmax values of 7.2 +/- 0.7 pmol/mg membrane protein and 11.4 +/- 1.0 pmol/mg membrane protein respectively. This indicates that there were probably two putative receptor sites for dip-allatostatin 7 although only one band was observable following photoaffinity labeling. Binding was saturable, specific and reversible. Using the in vitro binding assay, the Kd of the putative receptor in CA for dip-allatostatin 7 was shown to be (7.2 +/- 0.9).10(-10) M. PMID- 7480885 TI - Pseudodipeptide analogs of the pyrokinin/PBAN (FXPRLa) insect neuropeptide family containing carbocyclic Pro-mimetic conformational components. AB - Three N-terminal amino acid residues of the C-terminal core pentapeptide Phe-X Pro-Arg-Leu-NH2 (X = Gly, Ser, Thr, Val) of the pryokinin/PBAN insect neuropeptide family were replaced by nonpeptide moieties. To reestablish some of the conformational properties lost upon removal of the peptide bonds and Pro of the three amino acid residue block, carbocyclic Pro-mimetic components were incorporated into pseudodipeptide analogs. The most active analog contained a trans-DL-1,2-cyclopentanedicarboxyl carbocyclic component and proved to be over 3 orders of magnitude more potent than a simple, straight chain pseudodipeptide analog and approached the potency of the pentapeptide core in a cockroach hindgut myotropic bioassay. The pseudodipeptide analog retains a critical carbonyl residue which can participate in a hydrogen bond that stabilizes a beta-turn conformation in the active core region of the pyrokinin/PBAN peptides. This study demonstrates that knowledge of active conformation can be used to enhance the biological potency of pseudopeptide mimetic analogs of insect neuropeptides. The analogs represent a milestone in the development of pseudopeptide and nonpeptide mimetic analogs of this peptide family, which has been associated with such critical physiological processes as hindgut and oviduct contraction, pheromone biosynthesis, diapause induction, and induction of melanization and reddish coloration in a variety of insects. Mimetic analogs are potentially valuable tools to insect neuroendocrinologists studying these physiological processes and/or engaged in the development of future pest management strategies. PMID- 7480886 TI - Investigation of a screening battery for immunotoxicity of pharmaceuticals within a 28-day oral toxicity study using azathioprine and cyclosporin A as model compounds. AB - The authors have investigated a panel of parameters for immunotoxicity that may be incorporated in routine screening for toxicity of pharmaceuticals. This panel comprises serum immunoglobulin concentrations, cellularity of bone marrow, weights and histopathology of thymus, spleen, and lymph nodes, histopathology of Peyers' patches, and FACScan analysis of lymphocyte subpopulations in the spleen, in addition to parameters of toxicity to other systems. To study the value of these assays for pharmaceuticals, the authors used the immunosuppressants azathioprine (AZP) and cyclosporin A (CsA) as model compounds with known immunotoxic activity. In two separate experiments, rats were treated by daily gastric intubation with 0, 5, 12.5, and 25 mg AZP/kg body wt or 0, 1.25, 5, and 20 mg CsA/kg body wt. In the AZP study, the histopathology of the thymus and the spleen were valuable parameters of immunotoxicity, since these organs showed microscopic alterations at relatively low dose levels. In the CsA experiment, both the histopathology of the thymus and the data provided by FACScan analysis were sensitive indicators of immunotoxicity detecting effects at the lowest dose level employed. The data indicate that the lymphoid system is the most sensitive target of toxicity after AZP or CsA administration. The authors conclude that their test battery yielded immunotoxicity profiles of AZP and CsA in rats that were consistent with published findings in the literature, indicating the usefulness of the test battery employed. PMID- 7480887 TI - The ability of predicted internal dose measures to reconcile tumor bioassay data for chloroform. AB - PBPK models are developed in the hope that they will improve our ability to extrapolate from one species to another and from one exposure regime to another. Evidence that a dose measure was successful at reconciling the available animal bioassay data would be encouraging. It would give us some confidence that the dose measure (as evaluated by a PBPK model) might yield reasonable predictions for yet other species (e.g., humans) and other dose routes. We have investigated the ability of a modified version of the Corley et al. (R. A. Corley, A. L. Mendrala, F. A. Smith, D. A. Staats, M. L. Gargas, R. B. Conolly, M. E. Andersen, and R. H. Reitz, 1990, Toxicol. Appl. Pharmacol. 103, 512-527) PBPK model for chloroform to reconcile the available bioassay data. Two rate-dependent dose measures, maximal rate of metabolism in the liver, and percentage of hepatocytes killed per day performed well at reconciling the rodent liver bioassay data, while all rate-independent dose measures performed less well. In contrast, none of the PBPK dose measures were capable of reconciling the rat and mouse kidney tumor response data. Here, administered dose scaled to body surface area performed the best. PMID- 7480888 TI - A review of the acceptable daily intakes of pesticides assessed by WHO. AB - Over the past three decades, WHO has evaluated and reevaluated, through the Joint FAO/WHO Meeting on Pesticide Residues, 230 pesticides. The acceptable daily intakes (ADIs) of these pesticides are analyzed along with their scientific bases. In most cases, the evaluation process was consistent with the stated general principles and procedures of JMPR. However, the safety factors used in allocating the ADIs of several pesticides seem to be inconsistent with the severity of the toxicity, and thus, may require further consideration. These chemicals are abamectin, dinocap, procymidone, chlormequat, ethion, glyphosate, fentin hydroxide, and fentin compounds. In addition, pesticides that were evaluated many years ago, e.g., bromomethane, might merit a reevaluation in light of any relevant recent data. PMID- 7480889 TI - Assessment of novel foods: a call for a new and broader GRAS concept. AB - Novel or new foods may be those introduced for the first time from other parts of the world or they may result from the application of new or modified physical, chemical, and biotechnological treatments performed separately or combined. In addition to offering new possibilities of food for a growing human population, the development raises questions on the criteria for the acceptance of such foods. Conceptual, ethical, and technological issues are involved and call for a world-wide debate on a new and broader GRAS concept, including not only toxicological and nutritional elements but also cultural and philosophical values. PMID- 7480890 TI - Increasing lung cancer mortality rates in the elderly: a manifestation of differential survival. AB - Lung cancer mortality rates in the elderly are increasing. Using published United States mortality data, annual age-specific lung cancer mortality rates from 1968 to 1989 were determined for age groups over age 50 and compared to corresponding annual age group population sizes. Rising lung cancer mortality rates among the elderly in the United States from 1968 to 1989 were increasingly dependent, with increasing age, upon increasing age group population size. This finding suggests that differential survival, and its effect upon the surviving gene pool in an aging population, may account for observed increasing lung cancer mortality rates in recent successive elderly cohorts. That is, increasing lung cancer mortality rates in the elderly may reflect changes in the genetic susceptibility of the surviving population rather than changes in environmental exposures. PMID- 7480891 TI - Safety evaluation of pullulanase enzyme preparation derived from Bacillus licheniformis containing the pullulanase gene from Bacillus deramificans. AB - Pullulanase enzyme is an amylopectin debranching enzyme used in starch hydrolysis. This article describes studies conducted to investigate the safety of a pullulanase enzyme preparation produced by a strain of Bacillus licheniformis that has been transformed by introduction of genetic material from another Bacillus species, B. deramificans. A 4-week dietary toxicity study in rats was conducted in which test animals received pullulanase in the feed at concentrations of 0.2, 1.0, and 5.0%. No adverse treatment-related effects were observed. Lack of genetic toxicity potential was demonstrated by the results of a bacterial mutation assay in Salmonella typhimurium strains TA98, TA100, TA1535, TA1537, and TA1538, in an in vitro histidine forward mutation study in mouse lymphoma cells, and in in vivo mouse bone marrow chromosome aberration and micronucleus assays. The enzyme preparation also has been shown to be a nonirritant in eye and primary dermal irritation tests in rabbits and is nontoxic by inhalation exposure. Finally, the genetically altered B. licheniformis has been demonstrated to be nonpathogenic upon single intraperitoneal injection to rats of both live and killed cells at doses up to 10(11) cells/kg. The results of these studies demonstrate that the enzyme preparation may be considered safe when employed in starch processing. PMID- 7480892 TI - Alternative models for low dose-response analysis of biochemical and immunological endpoints for tetrachlorodibenzo-p-dioxin. AB - As part of the current reassessment of dioxin, the empirical relationships between administered tetrachlorodibenzo-p-dioxin and selected immune and biochemical endpoints were investigated. A dose-response analysis from the published literature was performed using Linear, Sigmoid-Emax and Power Law functions. The results of this analysis indicate that the use of a wide dose range may bias the interpretation of low-dose phenomenon. The Power Law function was applied exclusively to low-dose subsets enabling estimation of dose response in the low-dose range. Subsequently, high-dose data were fit using Power Law subset analysis. This approach resulted in a change in slope value from low- to high-dose subsets for thymic atrophy, immune suppression, benzo(a)pyrene hydroxylase activity, and ethoxyresorufin-o-deethylase activity. This change suggests that there is a high probability that there is a tissue concentration along the dose-response continuum which results in biological activity from low to high dose. This analysis also demonstrates that the Power Law functional fit to the low-dose data differs quantitatively from the fit to the high-dose data for several noncancer endpoints. Therefore, by using the Power Law function a more accurate and biologically relevant assessment of risk can be produced for non-cancer endpoints. PMID- 7480893 TI - Vermiculite: a review of the mineralogy and health effects of vermiculite exploitation. AB - Vermiculite is a mica-type mineral that is being used in increasing quantities for insulation, in composite cements, and in horticulture. No serious health risks have been found resulting from the exposure to vermiculite alone nor are any anticipated in view of its long-term chemical durability, even with respect to fibers of vermiculite. Vermiculite ores may contain a variety of other minerals including asbestos, which, if present in significant quantities, could pose a health risk to producers and end users. A variety of regulations are in place that should prevent inadvertent exposure to asbestos in vermiculites, but great care is needed if true asbestos hazards are to be correctly identified. At the same time, vermiculites containing any of a variety amphibole minerals should not be excluded from use because of mistaken identification of these minerals as asbestos. A variety of test methods that have been developed are described and recommendations are made with respect to appropriate action levels for asbestos in vermiculites or other raw materials. PMID- 7480894 TI - A model for estimating the daily dietary intake of a substance from food-contact articles: styrene from polystyrene food-contact polymers. AB - An approach has been developed to estimate the exposure of consumers to styrene from polystyrene food-contact articles which incorporates published literature on the diffusion of styrene through polymeric materials and industry survey data on uses of polystyrene in food-contact applications. The approach has been shown to be quite practical and has yielded an exposure estimate of 3 ppb (9 micrograms/day) styrene, on average, in the daily diet of people in the United States. This value compares reasonably well with the value of 1-4 micrograms/day for residents of the United Kingdom in 1983. The value is also four orders of magnitude less than the acceptable daily intake calculated by the Styrene Information and Research Center. PMID- 7480895 TI - Principles underlying dose selection for, and extrapolation from, the carcinogen bioassay: dose influences mechanism. AB - The purpose of the bioassay is not to simply find chemicals that can be labeled as carcinogens. On the contrary, the overall goal is to provide a reasonable assessment of the possible hazard that a chemical might pose to people under realistic conditions of exposure. This paper focuses upon the doses commonly used in the bioassay within the context that dose influences mechanism and, over a wide range of doses, mechanism changes with changing dose. Thus, a carcinogenic effect observed at a high dose is not necessarily expected to occur at lower doses. A variety of examples are provided to illustrate the points that (a) any high dose, no matter how high, that permits test animals to live long enough to develop tumors is not an appropriate criterion for defining an acceptable high dose to employ in a carcinogen bioassay; and (b) emphasis should be placed upon research that may discern probable thresholds for the carcinogenic effect of chemicals, especially nongenotoxic chemicals. PMID- 7480896 TI - FEMA GRAS--a GRAS assessment program for flavor ingredients. Flavor and Extract Manufacturers Association. AB - The generally recognized as safe (GRAS) assessment program of the Flavor and Extract Manufacturers Association (FEMA) of the United States was initiated in 1959 to provide for the assessment of flavor ingredients as GRAS under the Food Additives Amendment to the U.S. Federal Food, Drug, and Cosmetic Act. FEMA sponsored the formation of an independent panel of experts to perform GRAS assessments and to provide their conclusions to the U.S. Food and Drug Administration, the food and flavor industries, and the public. The program was designed to account for the legal, regulatory, and scientific issues associated with GRAS assessments and has continued to incorporate changes in the law and in science. This review describes the legal and scientific foundation of the FEMA program. PMID- 7480897 TI - Risk assessment: the default conservatism controversy. AB - EPA cancer risk assessment rests heavily on defaults. Defaults are a reduction of science to generic principles selected as a policy matter on the basis of "conservatism" for use in risk assessment. Conservatism is understood to mean a choice to avoid underestimating risk. The recent report of the National Academy of Sciences (1994) has turned the spotlight on the controversy regarding the use of generic principles as defaults and whether conservatism is the appropriate value criterion for their selection. Defaults had their origin in the early 1970s and the debate has continued regarding the scientific basis for the defaults and whether a conservatism, a value that the NAS said is "beyond science," is appropriate as a basis for the policy choices. This paper briefly examines the CAPRA recommendations to reduce the reliance on defaults, the history of the default conservatism controversy, and EPA's initial draft response to the CAPRA recommendations. PMID- 7480898 TI - [The diagnostic and therapeutic unity of endoscopic retrograde cholangiopancreatography in patients with a diagnosis of choledocholithiasis and its complications]. AB - Reviewing the records of 60 patients admitted with clinical diagnosis of common duct stone, we demonstrated that the symptoms and the biochemical liver tests were predictive of choledocholithiasis only 50% of the time. The sensitivity for the ultrasound was 42%. The ERCP diagnosed pancreatic or biliary pathology in over 90% of the patients. Through endoscopic sphincterotomy, the ERCP prevented the surgical exploration of the common bile duct in 93% of the patients, and did not aggravate the clinical picture in the group with acute pancreatitis. The sequence of ERCP--laparoscopic cholecystectomy decreased the hospital days and has emerged as the treatment of choice for the patients with cholelithiasis and suspected common duct stone. PMID- 7480900 TI - [Perinatal factors which affect low birth weight]. AB - The incidence of low-birth weight babies in the Santo Tomas maternity ward was 9% between the first of August of 1992 and the 31 of July of 1993. We found a higher risk of having low birth weight babies if the mother was toxemic, without prenatal care of low socioeconomic status and if she was single. We did not find an association with adolescence, short mother's height, low educational status and primi- or multiparity of 5 or more. PMID- 7480899 TI - [Adverse reactions to contrast media for intravenous use. A comparison between ionic and nonionic media]. AB - The purpose of this study was to show that the nonionic contrast media of low osmolarity are better tolerated than ionic media of high osmolarity, which to date are most frequently used in the authors hospital. We found that adverse reaction occurred in 80% of the group in which nonionic contrast medium was used. The authors conclude that nonionic, low osmolarity contrast media significantly reduce the frequency of adverse reactions and increase their safe use. PMID- 7480901 TI - [Subcostal, 3-4 cm, mini-cholecystectomy. A 3-year follow-up]. AB - The authors report the results of mini-cholecystectomy performed through a 3 to 4 cm long subcostal incision in 29 patients with the diagnosis of acute or chronic cholecystitis, from February 1991 to November 1922. Some of the patients were obese, diabetics or presented as emergency cases. The patients were operated on in the morning, as in laparoscopic cholecystectomy, began oral intake in the afternoon and were discharged on the day after surgery. Dissection of the gallbladder was facilitated by the use of a modified gynecologic valve and long thin instruments. Duration of surgery varied from 40 to 140 minutes. Patients could return to work on the third day after surgery. Notably, the costs/benefits were on the third more favorable than those of laparoscopic cholecystectomy. PMID- 7480902 TI - [Penetrating cardiac trauma]. AB - The author reviewed the clinical records of 40 patients with the diagnosis of penetrating cardiac trauma operated on in a period of 38 months in order to confirm the prognostic value of trauma indexes: Physiologic Index, Revised Trauma Scale and Index of Penetrating Cardiac Trauma. Thirty-eight patients were men and two were women with an average age of 26.8 years. Thirty-one patients suffered knife wounds and nine were wounded by bullets. The 15 unstable patients (systolic Blood Pressure 80 mmHg after infusion of crystalloid solutions) were submitted to thoracotomy. The 25 stable patients underwent the performance of a pericardial (subxiphoid) window for diagnosis, followed by sternotomy if the result was positive. There were 5 deaths all of which occurred in the operating room due to exsanguination. In all these cases Revised Trauma Scale was less than 3 and the Physiologic Index was grade III in four and grade II in one patient. Our results indicate that the Revised Trauma Scale and the Physiologic Index are good prognostic indicators of survival and that patients with Physiologic Index grade I and II have good probabilities for survival if they are operated on promptly. PMID- 7480903 TI - [Pseudosarcoma botryoides during pregnancy]. AB - Pseudosarcoma botryoides is an infrequent vaginal tumor with benign evolution. Its importance lies in to distinguish of sarcoma botryoides, what is usually fatal. A case of vaginal polyp during pregnancy is presented. After cesarean section, polyp was excised and diagnosis of sarcoma botryoides was made. Patient was referred to ION for treatment and her diagnosis was changed to pseudosarcoma by revision of histologic plates. Patient doesn't need further treatment but previous excision, and she is asymptomatic twelve months later. Histologic findings of both tumors are described and confusion between them are noted. PMID- 7480904 TI - [Breast cancer]. AB - The authors reviewed the clinical records of 31 patients with carcinoma of the breast diagnosed at the Dr. Rafael Hernandez Hospital in Chiriqui from January 1988 to December 1990. All the patients were women and the average age was 51.9 years. 41.9% of the patients were classified Stage 1; 29%, Stage 2; 16.1%, Stage IIIA; and 12.8%, Stage IIIB. 74% of the patients were treated with a modified radical mastectomy, radiation and hormonal therapies. Systemic cytotoxic therapy was given to 77% of the patients. Total survival 48 months after diagnosis was 83.3% in patients at Stage I; 66%, at Stage II; 40%, at Stage IIIA; and 0%, in patients at Stage IIIB. PMID- 7480905 TI - [Herpes zoster in infants]. AB - The authors present the clinical history of four children under two years of age who were hospitalized in the Arnulfo Arias Madrid Medical Complex with the diagnosis of herpes zoster. The mothers of these children had varicella when in the third, sixth, eight and fifth month of pregnancy respectively and the children were 3, 24, 14 and 8 months old when they had herpes zoster. The first child (whose mother had varicella when she was three months pregnant) was born underweight, dysphagic and premature. The fourth (whose mother had varicella in the 5th. month of gestation) was only underweight. The other two (mothers had varicella in the 6th and 8th month of pregnancy, respectively) were born without apparent abnormality. The authors, based on their findings, believe that there is risk for the child to have a congenital malformation when the mother develops varicella in the first months of pregnancy. PMID- 7480906 TI - [Optimization of the immunoelectrophoresis technic (western blot) for the confirmation of human immunodeficiency virus infection (HIV) in Panama]. AB - The purpose of this study is to report the results of the authors' investigation to apply the western blot technique (WB UP-LCS) in the diagnosis of human immunodeficiency virus type 1 (HIV-1) infection. To do this, the authors separated the proteins of the HIV-1 virus by electrophoresis, based on their molecular weight, in poliacilamide gel with SDS (SDS-PAGE) during 3 hours at 200 volts. Then they electrotransferred these proteins to nitrocellulose paper during four hours at 200 milliamperes, with the aid of external cooling. The nitrocellulose strips were evaluated considering the incubation time (1 and 16 hours), two conjugates (human anti IgG with Peroxidase and human anti IgG Biotin plus Streptatividine with Peroxidase) and two dilutions of the patients' sera (1/50 and 1/100). Based on their results the Authors conclude that, in the first place, the optimal conditions for the test include a dilution of 1/100 of the patients serum, incubation of the serum for 16 hours and the use of the conjugate of anti human IgG with Biotin and Streptavidine with Peroxidase; secondary, that the immunologic reactivity against proteins p24 and gp 160/120 is the most important diagnostic criterion for the confirmation of infection with HIV-1 and that they obtained a diagnostic correlation of 100% at a cost which was 5 to 7 times less than that of the commercial system. PMID- 7480907 TI - [Acute fatty liver in pregnancy. A diagnosis to be considered]. AB - The clinical records and follow up of two patients with jaundice in the third trimester of pregnancy and final diagnosis of acute fatty liver of pregnancy (AFLP) are reported. The purpose of this paper is to orient clinicians on the appropriate diagnostic approach in every woman with jaundice in the third trimester of pregnancy. PMID- 7480908 TI - Treatment of leprosy with WHO's standard fixed duration polychemotherapy. PMID- 7480909 TI - The hemorrhagic syndrome of leptospirosis: an experimental study in guinea pigs. AB - The hemorrhagic syndrome of leptospirosis was studied in guinea pigs. The study correlates hematological, histopathological and immunohistochemical alterations in sixty animals inoculated by the intraperitoneal route with 1ml of the culture of virulent strain of Leptospira interrogans serovar copenhageni. Leptospirae antigens were detected by immunoperoxidase, chiefly in liver, kidney and heart muscle capillaries. Possible pathogenic mechanisms responsible for hemorrhagic syndrome are discussed with emphasis on toxic and anoxic attacks causing damage to endothelia, platelet depletion and alterations to hemostasia rates: prothrombin time [PT], partial thromboplastin time [PTT] and fibrinogen concentrations. The clinical-laboratory picture is compatible with the histopathological observation of disseminated intravascular coagulation [DIC] in most of the guinea pigs from day 4 of infection. PMID- 7480910 TI - Antischistosomal activity of acridanone-hydrazones in Cebus monkeys experimentally infected with the SJ strain of Schistosoma mansoni. AB - In this study, four compounds were utilized at the dose of 12.5 mg/kg body weight, p.o., to treat Cebus monkeys experimentally infected with about 200 cercariae of Schistosoma mansoni (SJ strain), via transcutaneous route. The oograms performed with rectal snips, as well as stool examinations carried out periodically, showed no viable eggs of the parasite, from day 29 to 226 post treatment. The perfusion undertaken after killing the animals showed absence of worms in the treated monkeys, whereas 83 worms were recovered from the control, thus corroborating the results obtained by means of oograms and coproscopy. These results confirm the efficacy of 9-acridanone-hydrazones previously tested against the LE strain of S. mansoni. The low curative dose and apparent absence of toxicity render these drugs an important therapeutic reserve, taking into consideration the reports on the resistance of S. mansoni to the modern drugs oxamniquine and praziquantel. PMID- 7480911 TI - [Epidemiologic study on tegumentary leishmaniasis in the municipality of Corguinho, Mato Grosso do Sul -- Studies in the human population]. AB - A study on the resident population of 150 inhabitants of Boa Sorte in the Municipality of Corguinho, Mato Grosso do Sul State, Brazil was made, from March 1991 to March 1994, to establish the prevalence of South American Cutaneous Leishmaniasis (SACL), and to characterize the affected population, in an area of recent transmission. Twelve of the inhabitants showed lesions suspected to be SACL, and in 8 cases it was possible to confirm this by biopsy and parasitology. The mucosal form was found in one patient only, the rest showed the following cutaneous forms: ulcerated (3), ulcero-verrucose (1), hyperkeratotic ulcer (1), infiltrated maccule (1), nodule with florid regional adenopathy (1). All patients reacted favorably to treatment with glucantime, with lesion scarring. Side effects were rare. The parasite isolated from all patients was identified as Leishmania (Viannia) braziliensis. The Montenegro skin test, applied to the 150 inhabitants, showed 32 reactive ones. Of these, six were carriers of the disease, 21 showed sequelae suggestive of the disease and five showed no signs of infection. The age grouping of the cohort ranged from 22 to 78 years, 75% being male. To date, transmission is suspected to be in the peridomicile. PMID- 7480912 TI - [Feeding and defecation dynamics of Triatoma sordida]. AB - Today, Triatoma sordida is the most frequently captured triatomine in Brazil. For a better understanding of its vectorial potential, its feeding dynamics and dejections behaviour were studied in comparison with Triatoma infestans and P. megistus. The proportion of T. sordida and T. infestans that blood fed on anaesthetized rats did not differ significantly. There was no significant difference in the time elapse between release of the bugs and the initiation of blood feeding. The mean time between completion of a blood meal and the first dejection was significantly lower in T. infestans than in T. sordida. The numbers of insects that defecated during the blood meal or immediately afterwards was similar for both species. Approximately 80 specimens of each of T. sordida, T. infestans and Panstrongylus megistus were offered daily blood meals for 30 minutes, over a period of 30 days. T. sordida bit more frequently than did P. megistus and T. infestans. These data could indicate that T. sordida may not be completely adapted to its host and might have difficulty in complete its blood meal. PMID- 7480913 TI - [Seroepidemiology of hepatitis A virus infection in street children of Goiania Goias]. AB - A cross-sectional study was performed to determine the age-prevalence of hepatitis A virus (HAV) among street youth from Goiania city--Central Brazil, and to determine if any risk factors could be identified. The surveys were conducted between 1991/1992. The street youth were 397 individuals aged 7 to 21 years old living in institutions and teenagers working at streets. Then, 313 adolescents working at streets and with family links, and 84 institutionalized ones without family were screened for antibodies against HAV. Prevalences ranged from 80.0% to 92.2% to total anti-HAV and there was not a statistically significant trend of positivity with age in this group. Univariate analysis for risk factors associated with HAV infection was performed with no statistically significant difference for adolescents working at streets and living with parents, and street youth with family links. Economic variables were not statistically associated with seropositivity, probable due to homogeneous characteristics of the group. This study disclosed that street youth have a high rate of HAV infection. These findings do not suggest differences between subgroups of populations in acquiring immunity to HVA. The public health implication and the need of screening other subgroups of population of the same city were suggested in order to discuss vaccine strategy in underdeveloped countries. PMID- 7480915 TI - [Epidemiology of Toxoplasma gondii infection in the municipality of Ribeirao das Neves, MG. I. Importance of domestic animals as sources of T. gondii infection in humans]. AB - During a six month period (June 1983 to January 1984), a case-control study was carried out to determine if the variables associated with domestic animals contact would be correlated with the prevalence of Toxoplasma gondii infection. A randomized sample of 500 subjects from the population of Ribeirao das Neves, Minas Gerais, Brazil was studied. A statistically significant correlation was found with cat, chicken and pig contact but none with consumption of meat, milk and eggs. PMID- 7480914 TI - [Retrospective study on Latrodectus stings in Bahia, Brazil]. AB - This work is a retrospective study of latrodectism in the State of Bahia, Brazil, from August 1980 to July 1990. The data concerning the accidents were obtained from file cards at the Antivenom Information Center of Bahia (AVICB). Latrodectus curacavienis was the ethiologic agent identified in 28% of the arachnid accidents. The major incidence was registered in urban area (57%) affecting men (70%) more than women, with 10 to 29 year-old age group (58%). Local pain (56%), erythematous papula (29%) and light oedema (17%) were the principal local symptoms. Pain in the limbs (29%), tremor and rigidities (29%), sweating (28%), limbs and arms paresthesia (21%) and abdominal pain (17%) were systemic ones. The treatment was mainly symptomatic (67%) and antivenin serum was used in 21% of the cases. After serotherapy, 64% of the patients left the hospital within less than 24 hours. PMID- 7480916 TI - [Phlebotomus fauna (Diptera: Psychodidae) of 3 provinces of the northeastern region of Maranon, Peru]. AB - Two thousand seven hundred and seventy four sand flies were captured since 1987 to 1992 in endemic areas of human bartonellosis and Peruvian Verruga (district of San Jose de Lourdes, province of San Ignacio - 05 degrees 03' LS, and district of Lonya Grande, province of Utcubamba - 06 degrees 05'30" LS) belonging to the Northeast region of Maranon, Peru. Thirteen species were identified, 11 belonging to the genus Lutzomyia, 1 to Brumptomyia and the other to Warileya. Lutzomyia sp, close to L. serrana and L. maranonensis n. sp., Galatti, Caceres & Le Pont are more prevalent (89.74% of the individuals captured). Suggests working of the kind. Lutzomyia verrucarum (Townsend, 1913) is reported in Utcubamba province for the first time. PMID- 7480917 TI - [Participant survey in the intervention of intestinal helminthiasis transmission (satellite city of Paranoa -- Federal District]. AB - This work was developed in the Paranoa city town of the Federal District of Brazil. It used techniques of participative research with the aim of reducing intestinal helminths transmission in a group of children at nutritional risk. The family nucleus, that is, parents, children and other relatives living together in the same house, was used as reference for this research. All steps of the work were followed simultaneously by technical and educative actions. The technical actions were related to faecal examinations and medication. The educative process, stimulated critical analysis and reflections on the central problem and life conditions, with a strong focus on Health Promotion to improve life quality, as well as an empowerment process. During the work development, of the action program included the following steps: home visits, interviews, group meetings, feedback on the faeces examinations results and treatment. An evaluation of the final results was made through quantitative and qualitative methods, which showed the construction, acquisition and socialisation of knowledge. PMID- 7480919 TI - [Intestinal parasite contamination of vegetables sold at supermarkets in the city of Rio de Janeiro]. AB - Parasitological examinations were performed in material derived from 220 vegetables to detect cysts and eggs of intestinal parasites. This material was collected from supermarkets in northern and southern areas and from CEASA (Central Supply Market--40 vegetables) in Rio de Janeiro city. Vegetables proceeding from the northern areas presented a greater degree of contamination mostly of helminth eggs; lettuce was the most contaminated vegetable. The authors suggest that differences found between the northern and southern areas of the city may be due to either packing or manipulation by shop assistants and consumers. PMID- 7480918 TI - [Assessment of the Mitsuda reaction in lepromatous patients inactive before and after immunotherapy]. AB - In this study the immunopotentiator levamisole as well as a mixture of BCG/Mycobacterium leprae were investigated in inactive lepromatous leprosy patients by using the Mitsuda reaction as a parameter. Twenty lepromatous patients ten years ago classified as histologically negative for Mitsuda's test were divided into three groups: five patients that were only retested with Mitsuda antigen; eight patients that received oral levamisol and seven patients that received a mixture of alive BCG plus autoclaved M. leprae. The results indicated that: 1) the levamisole did not alter the reactivity to lepromin in any of the patients studied; 2) neither the changes in the reactivity to lepromin by using the mixture (3 cases) nor those that occurred spontaneously (3 cases) were clear. They properly reflected the natural variation of patients with some degree of resistance to Mycobacterium leprae. PMID- 7480920 TI - [American tegumentary leishmaniasis in the northeastern state of Sao Paulo Brazil]. AB - An outbreak of American Cutaneous Leishmaniasis (ACL) occurring in the Northeast region of Sao Paulo state in 1992 is described. After the notification of the 12 human cases, a skin test survey of the local population was carried out in a rural area of the Itupeva municipality. The survey consisted of 144 interviews and 100 clinical examinations using the Montenegro skin test (MST). A prevalence of 34% positive MST was encountered. The predominant species of sandflies captured both in domestic and nearby areas of secondary vegetation were L. intermedia, L. whitmani and L. migonei. The presence of L. longipalpis in the nearby area of secondary vegetation was also registered. PMID- 7480921 TI - [Delayed hypersensitivity to Trypanosoma cruzi antigen. I -- Experimental study in rabbits]. AB - Delayed-type skin hypersensitivity in rabbits infected with homologous or heterologous T. cruzi has been elicited upon injection of 50 micrograms of protein in the T12E antigen derived from a parasite clone. The specificity of this reaction was indicated by absence of skin reactivity in control rabbits that received same quantity of antigen. Also, the intradermal injection of the antigen in five occasions in control rabbits neither induced seroconversion nor shifted their skin sensitiveness. On the other hand, chagasic rabbits that underwent a series of five skin testings one week apart, did not alter the intensity of the skin reactivity, and their specific serum antibody titers remained similarly high. Of interest, the ECG patterns remained unchanged after a series of five skin testings in control and in chagasic rabbits. This study shows that the T12E antigen might be an useful tool for diagnosing T. cruzi infections, besides serving as an immunologic marker for morbidity in Chagas' disease. PMID- 7480922 TI - [Delayed hypersensitivity to Trypanosoma cruzi antigen. II--Use of the skin test with T12E antigen for the diagnosis of Chagas disease]. AB - Delayed-type skin reactivity against a Trypanosoma cruzi antigen was elicited in 35.7 percent of the individuals living in the country of Mambai, state of Goias, Brazil. The specificity of this skin reaction was shown in 93 out of 94 (98.7%) chagasic patients, in whom the parasitemias were detected by xenodiagnosis. In these patients, however, the hemagglutination, immunofluorescence and complement fixation assays were positive in respectively, 100, 97.8 and 80.6 per cent. The relationship between the combined positive results by hemagglutination and immunofluorescence with that obtained with skin testings was 0.897, in the overall population in this study. The quantity of 50 micrograms of protein in 100 microliters of the T12E antigen did not produce undesired effects, and did not shift the immunologic assays, when healthy volunteers were skin tested five times within 15-day intervals. Also, the potency of this antigen remained unaltered after 24 months at -10 degrees C. PMID- 7480923 TI - [Delayed hypersensitivity to Trypanosoma cruzi antigen. III--Sensitivity of the skin test with T12E antigen in the diagnosis of Chagas disease in hospitalized patients]. AB - In this study we evaluated three immunologic methods for confirmation of clinical diagnosis of Chagas' disease, in 120 in-patients. The skin testing with the T12E antigen and the immunofluorescence were positive in 10% of cases. The hemagglutination was positive in 14.1% of these patients. The co-positivity of results of skin-testing with hemagglutination and of this latter with immunofluorescence reached 7.5% of cases. However, only 5% of co-positivity was achieved with results of skin testing, immunofluorescence and hemagglutination. Furthermore, 19.1% of the in-patients showed at least one out of these exams to be positive. The specificity of the skin testing paralleled that of the immunofluorescence. The sensitivity of these exams, however, was lower than that obtained by the indirect hemagglutination. These data show that the skin testing with T12E antigen allows the diagnosis of Chagas' disease by a simple and handy method. PMID- 7480924 TI - [Effectiveness of aminosidine sulphate in severe visceral leishmaniasis, resistant to the treatment with pentavalent antimony]. AB - A grave kala-azar infection in a 14 years old boy is described. The leishmanial infection failed to respond to ten interrupted courses of glucantime of variable duration (14-56 days) at a dose of 20mg Sb5/kg/day. However a favorable response occurred to intramuscular aminosidine sulphate (20mg/kg/day) for 20 days. This same regimen was repeated 20 days later. After the first treatment splenic puncture parasite density fell from 50 amastigotes per oil immersion field to 3 amastigotes in 10 fields. A further splenic puncture 7 months after treatment was negative. The marked hepatoesplenomegaly gradually resolved over 26 months follow up and he gained 13 kilograms in weight. After aminosidine sulphate therapy his Montenegro reaction become positive and his lymphocytes responded to leishmania antigens. PMID- 7480925 TI - [Paracoccidioidomycosis in a patient with human immunodeficiency virus infection. A necropsy case]. AB - This is a case report of the association of Paracoccidioidomycosis and Acquired Immunodeficiency Syndrome (AIDS) occurring in a 43-year old male. This is, to the best of our knowledge, the first detailed pathological account of that association. Also discussed are the low rates of that association, its natural history and treatment results. It is emphasised the importance of the associations of AIDS and tropical infectious diseases in this country. PMID- 7480926 TI - [A new case of human pulmonary dirofilariasis acquired in Brazil]. PMID- 7480928 TI - [Decision to extract for periodontal reasons]. AB - The decisions to extract that are discussed here mainly relate to severe periodontal bone losses and plain interradicular attacks. Extractions for periodontal reasons appear at several treatment stages: immediate extraction in terminal stages, e.g. for teeth without any strategic importance, or deferred extraction if one chooses to wait for treatment response. The elements of the decision are various: function and strategic importance of the tooth, extension of the lesion, level of the interradicular attack, any endoperiodontal lesions, fractures and luxations, radicular proximity, implication of wisdom teeth, evolution after treatment. All these factors are discussed. PMID- 7480927 TI - [A case of urticaria during pentavalent antimony use in a patient with mucous leishmaniasis]. PMID- 7480929 TI - [Should we or should we not extract impacted teeth?]. AB - Do we have to remove the impacted teeth? The pathology of an impacted tooth is reviewed by the author as well as the possible complications of its removal and others treatment possibilities. Seeing the increase of pathology and operative complications owing to the age, the author propounds the management of immature impacted third molars when they are not in a propitious position or when the posterior eruption space is reduced. About the impacted teeth covered partially or completely by soft tissue, his advice consists too in removing them if they don't have the possibility to take up good position on the dental arch. PMID- 7480930 TI - [The decision to extract in orthodontics]. AB - Serial extractions of premolars are submitted, in the present state of the contemporary orthodontics, in opposition to the non-extraction treatments (maxillary expansion, molar distancing...). The choice depends on many parameters including the severity and the nature of the malocclusion, the age and the morphological type of the patient and also the "orthodontic philosophy" relating to the techniques used by each orthodontist. The indications and contraindications of these teeth removals are explained showing the benefits and the disadvantages in different cases. "Orthodontics extractions" of other teeth (2nd bicuspid, 1st and 2nd molar, incisor) are also discussed. PMID- 7480931 TI - [Extraction in orthodontics: when and why?]. AB - During the last century orthodontic treatment philosophies changed continuously. The necessity to extract teeth in order to correct a lack of space has many times been the subject of discussion. In fact there exists several techniques which make it possible to avoid extractions in many cases. A number of criteria which influence the decision to extract or not, as well as some techniques to regain space in upper and lower arch are discussed. PMID- 7480932 TI - [Guided tooth eruption via serial extraction]. AB - Crowding of teeth is the result of arch length discrepancy (ALD). Minimal crowding can be solved with slice-technique, more severe crowding (ALD > or = 5 mm) can be intercepted with a serial extraction procedure. In this paper a sequence of serial extraction will be explained and illustrated with case reports. PMID- 7480933 TI - [The decision to extract]. PMID- 7480934 TI - [The limits of conservative care in pedodontics]. AB - Despite progress in the decrease of caries by means of many preventive dental programs, children's teeth of poor socio-economic groups are still strongly affected. The well known role of children's teeth in oro-facial growth and harmony, as well as in the development of functions as chewing, speaking or phonation and swallowing, has justified their preservation during all their functional periods. Several conservative therapies allow to reach this purpose. However, extensive dental caries, difficulty to get child tooth infection under control, injuries on permanent tooth germs and child's general health deficiency, can limit preservation of infected teeth, leading to their extraction. PMID- 7480935 TI - [Effects of Casimiroa edulis (Rutacea) on blood pressure and heart rate in albino rats]. AB - Aqueous extracts of Casimiroa edulis leaves injected intravenously to 20 normal Sprague-Dawley rats produced a rapid and transitory increase in blood pressure. The amplitude of the blood pressure rise was dose dependent. The increase was statistically significant (p < or = 0.01) at a concentration of 220 mOsm/l, and was associated with a significant heart rate decrease (p < or = 0.01) probably secondary to barorreceptor stimuli. The application of 130 mOsm/l of the extract did not produce a significant increase in blood pressure, which was followed by a rapid pressure normalization. The slight increase in cardiac rate during the pressure lift was probably due to sympathetic stimuli. PMID- 7480936 TI - Seroepidemiology of Toxoplasma gondii (Apicomplexa) in meat producing animals in Costa Rica. AB - A serologic screening for Toxoplasma gondii antibodies by IgG indirect fluorescent antibodies test (IgG-IFAT) was carried among 496 swine and 601 cattle serum samples from throughout Costa Rica, to study the possible role of their meat in the transmission of the parasite. The overall prevalence of antibodies was 34.4% in cattle and 43.8% in swine. No significant differences were found in the antibody prevalence between males and females in both animal groups, which acquire the infection early in their lives. Swine did not present significant differences between age groups. In cattle there was a high percent of seropositivity between the first and third years of age. The provinces that showed a greater number of seropositive animals were Limon and Puntarenas for both animals, and Guanacaste for cattle. PMID- 7480937 TI - [Reproduction and average size at maturity of the trout Oncorphynchus mykiss (Salmoniformes:Salmonidae) in the Venezuelan Andes)]. AB - From January 1987 to February 1988, a total of 393 rainbow trout were captured in a high mountain stream of the Venezuelan Andes (8 degrees 40'N; 71 degrees 5'W). Gonadal development was measured using the Nikolsky method. The mean size for sexual maturation was analyzed with the method of relative frequency of adults and young for each size class. Males reproduced throughout the year, while ripe females were observed only in January (11%). A half of the males was sexually mature at the minimum size of 13.40 cm SL (16.25 for females). All individuals were sexually mature at 26.40 cm (males) and 26.80 cm (females). PMID- 7480938 TI - [Home range, use of habitat and daily activity of the gopher Orthogeomys heterodus (Rodentia: Geomyidae) in a Costa Rican horticultural area]. AB - Home range, daily activity and habitat use of four Giant Pocket Gophers Orthogeomys heterodus were determined by radiotelemetry. The study was carried out in a horticultural area of Cartago province, Costa Rica, during December 1989 and January 1990. Home range averaged 325 m2 for males and 233 m2 for females. There was a significant relationship between home range size and individual body mass. In average, 78% of localizations were in the nest or refuge located in all cases in the border of crops. Activity was greatest from 0600 to 0800 h and 1200 to 1400 h, and was reduced at night. PMID- 7480939 TI - Patterns of reproduction among the Fang of Nsork (Equatorial Guinea): pregnancies, abortions and child spacing. AB - Fang women are known to practice virtually no contraception but for them induced abortion is not an acceptable option. Their reproductive behaviour consequently is governed by the ability to conceive, spontaneous intrauterine mortality and child spacing (due to prolonged breast-feeding and sexual abstinence). In a sample of 587 women from one hospital and one clinic in Nsork, there was a positive correlation between maternal age and the number of pregnancies, resulting in a mean of 5.52 pregnancies per female and one child born every 2.5 years. The reported spontaneous abortion rate was 28.6%. PMID- 7480940 TI - [The effect neutron irradiation of Uranium 235 on the venom of Lachesis muta muta (Ofidia: Viperidae)]. AB - The effect of irradiation of the aqueous solution of L. m. muta venom was evaluated with thermic neutrons from Uranium-235 employing doses of 1.61 to 5.19 Gray. The venom was examined for protein content by the Folin Lowry Method modified by Stauffer; for acute toxicity by intraperitoneal route estimation in mice and for immunochemical tests by the antigen-antibody reactions evaluation. Neutronic radiation affects all evaluated parameters in venom (decrease in protein content levels, increase of LD50 values and decrease in the number of precipitating antigen-antibody) as shown by immunodiffusion and immunoelectrophoresis. PMID- 7480942 TI - Incidence of Listeria monocytogenes in pasteurized ice cream and soft cheese in Costa Rica, 1992. PMID- 7480941 TI - [Cicatrization effect of Jatropha curcas latex (Angiospermae: Euforbiaceae)]. AB - The wound healing effects of Jathopha curcas latex upon surgical wound produced in Balb/c mice skin, were studied with a modification of the Hoowes-Sooy-Harvey method. The effects of topical treatment using single 50 ul doses of latex at different dilutions (10% to 100%) was compared with a multiple dose treatment (four 25 ul/dose q12h, latex 5% to 100%). The single dose treatment with 10%, 50% or 100% latex and the multiple dose treatment with dilutions between 5% and 10%, have a healing effect but only on males. The multiple dose treatment with 50% or pure undiluted latex produced caustic lesions to treated skin. PMID- 7480943 TI - Cockroaches as transport hosts of the protozoan Toxoplasma gondii. PMID- 7480945 TI - The effects of community pluralism on press coverage of health risks from local environmental contamination. AB - Based on the conflict/consensus model of Tichenor, Donohue and Olien, we proposed that mass mediated information signalling that local agents are contaminating the local environment and posing health risks is conflict-generating information and, therefore, will be controlled in the interest of community stability. We expected such control to vary by community structure. A content analysis of nine months of coverage by 19 newspapers supported the hypothesis that papers in more pluralistic communities were more likely than papers in less pluralistic communities to link contamination from local agents to threats to human health in the community and to frame such stories as problems. Newspapers in less pluralistic communities were more likely to frame local contamination in the context of solutions to the problem and were more likely to link contamination to health risks if the contamination were in a distant community. PMID- 7480944 TI - [Isolation of the bacteria Ureaplasma sp. in the reproductive tract of milking cows in Costa Rica]. AB - This is the first report of Ureaplasma sp. from the reproductive tract of Costa Rican cows. Among 204 animals sampled from 11 dairy farms in the country's Central Plateau, the infection rate was 0-71%. Isolation was more frequent in vulvo-vestibular (38.7%) than in cervical swabs (23%). Ureaplasma was correlated with clinical granular vulvitis symptoms. PMID- 7480946 TI - An improved approximation to the exact solution of the two-stage clonal growth model of cancer. AB - Multistage clonal growth models are of interest for cancer risk assessment because they can explicitly incorporate data on cell replication. Both approximate and exact formulations of the two stage growth model have been described. The exact solution considers the conditional probability of tumors arising in previously tumor-free animals; the approximate solution estimates total probability of tumor formation. The exact solution is much more computationally intensive when time-dependent cell growth parameters are included. The approximate solution deviates from the exact solution at high incidences and probabilities of tumor. This report describes a computationally tractable, 'improved approximation' to the exact solution. Our improved approximation includes a correction term to adjust the unconditional expectation of intermediate cells based on the time history of formation of intermediate cells by mutation of normal cells (recruitment) or by cell division in the intermediate cell population (expansion). The improved approximation provided a much better match to the exact solution than the approximate solution for a wide range of parameter values. The correction term also appears to provide insight into the biological factors that contribute to the variance of the expectation for the number of intermediate cells over time. PMID- 7480947 TI - Significance of exposure assessment to analysis of cancer risk from inorganic arsenic in drinking water in Taiwan. AB - The primary source of evidence that inorganic arsenic in drinking water is associated with increased mortality from cancer at internal sites (bladder, liver, lung, and other organs) is a large ecologic study conducted in regions of Southwest Taiwan endemic to Blackfoot disease. The dose-response patterns for lung, liver, and bladder cancers display a nonlinear dose-response relationship with arsenic exposure. The data do not appear suitable, however, for the more refined task of dose-response assessment, particularly for inference of risk at the low arsenic concentrations found in some U.S. water supplies. The problem lies in variable arsenic concentrations between the wells within a village, largely due to a mix of shallow wells and deep artesian wells, and in having only one well test for 24 (40%) of the 60 villages. The current analysis identifies 14 villages where the exposure appears most questionable, based on criteria described in the text. The exposure values were then changed for seven of the villages, from the median well test being used as a default to some other point in the village's range of well tests that would contribute to smoothing the appearance of a dose-response curve. The remaining seven villages, six of which had only one well test, were deleted as outliers. The resultant dose-response patterns showed no evidence of excess risk below arsenic concentrations of 0.1 mg/l. Of course, that outcome is dependent on manipulation of the data, as described. Inclusion of the seven deleted villages would make estimates of risk much higher at low doses. In those seven villages, the cancer mortality rates are significantly high for their exposure levels, suggesting that their exposure values may be too low or that other etiological factors need to be taken into account. PMID- 7480948 TI - Presenting uncertainty in health risk assessment: initial studies of its effects on risk perception and trust. AB - Some analysts suggest that discussing uncertainties in health risk assessments might reduce citizens' perceptions of risk and increase their respect for the risk-assessing agency. We tested this assumption with simulated news stories varying simple displays of uncertainty (e.g., a range of risk estimates, with and without graphics). Subjects from Eugene, Oregon, read one story each, and then answered a questionnaire. Three studies tested between 180 and 272 subjects each. Two focus groups obtained more detailed responses to these stories. The results suggested that (1) people are unfamiliar with uncertainty in risk assessments and in science; (2) people may recognize uncertainty when it is presented simply; (3) graphics may help people recognize uncertainty; (4) reactions to the environmental problems in the stories seemed affected less by presentation of uncertainty than by general risk attitudes and perceptions; (5) agency discussion of uncertainty in risk estimates may signal agency honesty and agency incompetence for some people; and (6) people seem to see lower risk estimates (10(-6), as opposed to 10(-3)) as less credible. These findings, if confirmed, would have important implications for risk communication. PMID- 7480949 TI - On the MVK stochastic carcinogenesis model with Erlang distributed cell life lengths. AB - This paper proposes extending the MVK carcinogenesis model by adopting the Erlang distribution for the life length of the intermediate cells. The investigation concentrates on the survival function and the mean value functions. The approach is basically numerical, making use of the Mathematica software system. The paper also provides a closed form expression for the survival function for a variation of the original MVK model, where all the model parameters are piecewise constants. PMID- 7480950 TI - Gender differences in risk perception: effects differ in stressed vs. non stressed environments. AB - Contrary to previous reports that women are more concerned about environmental risks than men, we hypothesized that men and women residing in neighborhoods stressed by multiple hazards would demonstrate similar concerns about local environmental conditions. Analysis of a national data base and an aggregate of ten local data bases found greater female than male concern about local technological, behavioral, and land use hazards in good neighborhoods, but, as expected, not in stressed ones. We urge analysts to conduct more studies in stressed neighborhoods in order to better understand the perspective of those who live with environmental risks. PMID- 7480951 TI - Perception of nuclear energy and coal in France and The Netherlands. AB - This study focuses on the perception of large scale application of nuclear energy and coal in the Netherlands and France. The application of these energy-sources and the risks and benefits are judged differently by various group in society. In Europe, France has the highest density of nuclear power plants and the Netherlands has one of the lowest. In both countries scientists and social scientists completed a questionnaire assessing the perception of the large scale application of both energy sources. Furthermore, a number of variables relating to the socio cultural and political circumstances were measured. The results indicate that the French had a higher risk perception and a more negative attitude toward nuclear power than the Dutch. But they also assess the benefits of the use of nuclear power to be higher. Explanations for these differences are discussed. PMID- 7480952 TI - Reanalysis of dose-response data from the Iraqi methylmercury poisoning episode. AB - Applying a hockey stick parametric dose-response model to data on late or retarded development in Iraqi children exposed in utero to methylmercury, with mercury (Hg) exposure characterized by the peak Hg concentration in mothers' hair during pregnancy, Cox et al. calculated the "best statistical estimate" of the threshold for health effects as 10 ppm Hg in hair with a 95% range of uncertainty of between 0 and 13.6 ppm. A new application of the hockey stick model to the Iraqi data shows, however, that the statistical upper limit of the threshold based on the hockey stick model could be as high as 255 ppm. Furthermore, the maximum likelihood estimate of the threshold using a different parametric model is virtually zero. These and other analyses demonstrate that threshold estimates based on parametric models exhibit high statistical variability and model dependency, and are highly sensitive to the precise definition of an abnormal response. Consequently, they are not a reliable basis for setting a reference dose (RfD) for methylmercury. Benchmark analyses and statistical analyses useful for deriving NOAELs are also presented. We believe these latter analyses- particularly the benchmark analyses--generally form a sounder basis for determining RfDs than the type of hockey stick analysis presented by Cox et al. However, the acute nature of the exposures, as well as other limitations in the Iraqi data suggest that other data may be more appropriate for determining acceptable human exposures to methylmercury. PMID- 7480953 TI - Tissue dosimetry, pharmacokinetic modeling, and interspecies scaling factors. AB - Interspecies scaling factors (ISFs) are numbers used to adjust the potency factor (for example, the q1* for carcinogens or reference doses for compounds eliciting other toxic endpoints) determined in experimental animals to account for expected differences in potency between test animals and people. ISFs have been developed for both cancer and non-cancer risk assessments in response to a common issue: toxicologists often determine adverse effects of chemicals in test animals and then they, or more commonly risk assessors and risk managers, have to draw inferences about what these observations mean for the human population. This perspective briefly reviews the development of ISFs and their applications in health risk assessments over the past 20 years, examining the impact of pharmacokinetic principles in altering current perceptions of the ISFs applied in these health risk assessments, and assessing future directions in applying both pharmacokinetic and pharmacodynamic principles for developing ISFs. PMID- 7480954 TI - Diagnostic-based pharmacological treatment of behavior disorders in persons with developmental disabilities: a review and a decision-making typology. AB - This article reviews the diagnostic rationale for treatment of behavior disorders with psychotropic medication. After reviewing the concerns over the use of psychotropic medication and the use of diagnostic criteria with persons with developmental disabilities in the past, the variety of potential diagnoses that may underlie a behavior disorder are reviewed. A model to guide clinical decision making is then developed on the basis of this review, and areas for future research are suggested. The key questions are identified as making reliable differential diagnoses of behavior disorders and demonstrating that these differential diagnoses successfully identify more effective psychotropic medications for the treatment of behavior disorders in persons with developmental disabilities. PMID- 7480955 TI - Therapeutic shock device (TSD): clinical evaluation with self-injurious behaviors. AB - A man with profound mental retardation and multiple topographies of severe self injurious behavior (SIB) had been receiving contingent shock for SIB for 2 years before the study started. Shock was being delivered with a handheld shock stick (Hot Shot Sabre Six), which produced burns to the man's skin. SIB rate, without a shock contingency, was 10 responses/min. The Therapeutic Shock Device (TSD), worn by the client and remotely operated by a radio frequency signal, provided superior control of SIB (0.02 responses/min) compared with the shock stick (0.06 responses/min) without causing tissue damage. TSD treatment was introduced in a mixed multiple baseline design across times, settings, and behaviors. The client did not appear to find the TSD aversive. To the contrary, his behaviors indicated that he preferred to wear it. The TSD appeared to provide a substitute for restraint, the hypothesized reinforcer for the man's most frequent form of SIB. PMID- 7480956 TI - Analog baselines: a critical review of the methodology. AB - Analog baselines are an experimental methodology for identifying the functions of maladaptive behavior in the naturally occurring environment (Iwata, Dorsey, Slifer, Bauman, & Richman, 1982; Iwata et al., 1994). This article identifies a number of potential limitations in this methodology. These include: (a) procedural problems inherent in the use of multielement designs, (b) the fidelity of analog baseline design conditions, (c) the relation of the analog conditions to the naturally occurring environment, (d) a narrow analysis of behavior limited by an implicit adherence to an ABC model of behavior, (e) a limited acknowledgement of multifunction and idiosyncratically motivated behaviors, (f) problems in the definition of response classes, and (g) difficulties in the use of analog baselines to design interventions. Future research should attend to three main questions. First, the convergent validity of different assessment methodologies, including analog baselines, should be evaluated. Future research should attend to procedures that can integrate the entire clinical process of referral, identifying the functions of the target behavior, including other methods of identifying the functions of behavior, treatment design, and implementation. Second, assessment failures could be examined carefully to identify ways of developing this methodology further. Third, the process of designing an intervention depends upon input from many sources of information. The use of analog baselines will be enhanced by a greater understanding of the process of clinical decision making. PMID- 7480957 TI - Major depression in a small group of adults with Down syndrome. AB - The clinical histories and treatment of the nine individuals with Down syndrome (DS) and major depression (MD) previously noted in a report on the psychopathology of a population of 164 adults with DS with and without health disorders from a Down Syndrome Clinic are presented (Myers & Pueschel, 1991). The clinical characteristics including DSM-III-R (1987) criteria of these 9 patients plus 13 individuals with DS and MD described in case reports in the literature are summarized. Depression is rarely verbalized and commonly appears as crying, depressed appearance, or mood lability. Vegetative symptoms of disinterest with severe withdrawal and mutism, psychomotor retardation, decreased appetite, weight loss, and insomnia are prominent. Verbal expression of preoccupations of suicide, death, self-depreciation, and guilt were infrequent and may either be not present or not reported due to mutism or moderate level of mental retardation (MR). Hallucinations were prominent. Family history of depression was infrequent. Psychological stressors were noted mostly in the study sample and not in the 13 from the literature. The pattern of vegetative symptomatology with few verbal complaints and prominent hallucinations may be related to moderate mental retardation in these groups with DS rather than specifically to DS. PMID- 7480958 TI - Descriptive analysis of multiple response topographies of challenging behavior across two settings. AB - Time-based lag sequential analyses were conducted on 23 topographies of challenging behavior shown by five young people with severe mental retardation across two settings. Potential behavioural functions were identified for 21 of the 23 behaviors. Responses classes, including two or more distinct behaviors, were identified for four of the five participants. Two participants showed evidence of two functionally distinct response classes; for one person each response class included both self-injurious and stereotypic behaviors. For 9 of the 11 behaviors for which data were available, the hypothesised function of behaviors was consistent across settings. For one person, data suggested that the function of one self-injurious behavior was contextually determined. Cross validation with brief experimental analyses resulted in agreement on the general behavioral function for 12 of the 14 instances in which both approaches positively identified a potential function for a behavior. Implications for future research and practice are discussed. PMID- 7480959 TI - Parental reports of community activity patterns: a comparison between young children with disabilities and their nondisabled peers. AB - The extent and nature of community activities of families of children with disabilities in comparison with their nondisabled peers were examined. Parents of 82 children with disabilities and 132 children without disabilities, ages 2 through 5 years, completed a Demographic Questionnaire, Community Activities Questionnaire (CAQ), and Parent Experience Survey (PES). The CAQ assessed frequency of child participation in age-appropriate community activities, and the PES was a retrospective self-evaluation by parents of the experience of introducing the child to common community settings. Both groups of parents reported highly similar experiences when shopping with their children, although parents of children with disabilities reported more fear and risk associated with public settings. Overall, parents of children with disabilities reported participation in fewer community activities than parents of children without disabilities. This difference was largely accounted for by a lower level of participation by families having children with disabilities in a subset of activities that reflect opportunities for family enrichment. PMID- 7480960 TI - [Cytomegalovirus infection in HIV]. AB - The data are taken from a retrospective study of 24 patients with AIDS who developed CMV disease. This study was performed to detect the principal characteristics associated with the development of CMV disease. Age, sex, risk factors, the time interval between HIV seropositivity to CMV disease were examined, as well as efficacy of gancyclovir therapy in both groups of patients with symptomatic and asymptomatic retinitis. When the chorioretinitis is still asymptomatic the early treatment of CMV disease may suppress progression of retinic lesions and seems to be efficacious to prevent the evolution of CMV disease. PMID- 7480961 TI - Atrial natriuretic factor and sympathetic activation in human heart failure. AB - The neurohormonal changes occur early, have important prognostic value, and may play a role in the evolution and progression of heart failure in man. Atrial natriuretic factor (ANF) is a natriuretic and vasorelaxant peptide. Previous studies indicated that plasma ANF provides prognostic information and, ANF levels closely related to both severity of disease and catecholamine levels but, it is still unclear if high circulating levels of ANF, which are present in heart failure constantly, may be to correlate with sympathetic nervous activity in man. Thus, the aim of the present study was to investigate the relations between the release of ANF and the sympathetic system in human heart failure. We studied 18 patients with heart failure (CHF) and a control Group (n = 14) of healthy subjects. To induce adrenergic activation in physiologic way patients were underwent to a low-exercise by cycle-ergometer in supine position. Blood was collected at rest, and immediately after exercise for determination of plasma levels of ANF, norepinephrine and epinephrine levels. ANF values at rest were 35.9 +/- 19.2 pg/ml in controls and 190.7 +/- 34.2 pg/ml (p < 0.001 vs controls) in CHF patients. As well norepinephrine levels showed higher values in patients (295.7 +/- 47.8 pg/ml), than in normal subjects (143.5 +/- 33.3 pg/ml; p < 0.01). In CHF patients epinephrine levels were 100.1 +/- 21.2 pg/ml (p < 0.01 vs controls). ANF levels were in normal subjects 87.9 +/- 19.2 pg/ml (p < 0.01 vs rest) after exercise. In CHF patients ANF values were 275.3 +/- 59.8 pg/ml; p < 0.001).(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7480962 TI - Theophylline in the treatment of the elderly with chronic obstructive pulmonary disease. AB - The authors describe the pharmacological effects of theophylline and elucidate its mechanisms of action according to latest hypothesis. The narrow therapeutic range of the drug and the existence of factors capable of modifying its distribution and clearance in the elderly, are taken into consideration, as well. The behaviour of theophylline concentrations in two groups of elderly subjects affected by chronic bronchitis, treated respectively with intravenous aminophylline and anhydrous theophylline administered orally are assessed. Usefulness of associating the two routes of administration is underlined. In fact, the delay in the achievement of the therapeutic range observed after oral intake can be made up for by the initial peak induced by intravenous injection. Our observations in the elderly are consistent with the necessity of therapeutic monitoring. In addition, a careful dosage of the drug can avoid the occurrence of untoward effects. PMID- 7480963 TI - [Long-term therapy in chronic hepatitis C with recombinant alpha interferon]. AB - The dosage and duration of Chronic Hepatitis (CH) C therapy with alpha (alpha IFN) is still an unresolved problem. Considering mainly antiviral activity of alpha-IFN, the authors carried out research on 35 patients with CH-C to evaluate the efficacy and safety of long term therapy and to compare recombinant alpha-IFN 2a with recombinant alpha-IFN-2b. 21 patients (Group A: 20 males, 1 female, mean age 48.8 years, 14.2% CPH, 76% CAH, 9.5% AC) were treated with r-alpha-IFN-2a; 14 patients (Group B: 7 males, 7 females, mean age 45.2 years, 7.1% CPH, 78.5% CAH, 14.3% AC) were treated with r-alpha-IFN-2b, at a dosage of 3 MU thrice weekly (TIW) for 12 months; the follow-up was from 6 to over 24 months. Complete Response (CR) was achieved in 49.3% of the patients treated, Long Term Response (LTCR) in 19.3%, Partial Response (PR) in 29% and No Response (NR) in 22.6%), Delayed CR in 6.5%. The authors found a similar number of CR and LTRC in group A and group B; a greater percentage of PR in group A and a greater percentage of NR in group B. Early and serious side effects in 4 patients, late and serious side effects in 2 patients were observed. The date suggest that the long term therapy of CH with r-alpha IFN is effective in inducing clinical and biochemical remission in a large portion of patients and a greater number of sustained response, with respect to a therapy schedule of 6 months even at greater dosages. PMID- 7480964 TI - [Reinfusion ascites therapy: considerations after a year's experience]. AB - Ascites often appears as a complication of several illnesses. The therapy is essentially based on the use of low-sodium diet, plasma or albumin infusion, diuretics and low-dosed ACE-inhibitors. To use the simple paracentesis or special techniques as Rhodiascit or Lee Veen Shunt means not to resolve definitively the problem and sometimes to cause undesirable complications. The authors present a new therapeutic tactics that joins the use of technique of double filtration of ascitic fluid and reinfusion of concentrated proteins (DFAF) with the injection in the peritoneal cavity of beta-interferon and the venous infusion of ATIII. Twenty patients affected by hepatic cirrhosis with the presence of ascitic fluid not treatable with the usual therapy have been subjected to this treatment. All the patients showed an immediate improvement of the clinical situation. After one year of observation, we describe our results. Twelve patients needed a further treatment with the DFAF technique, two patients died for the original pathology and six patients just needed an adjustment of pharmacologic therapy. PMID- 7480965 TI - Acute effects of alpha-1 adrenoceptor antagonist, doxazosin on circulating vasoactive hormones. AB - The acute effects of doxazosin, a new selective alpha-1 adrenergic blocking agent, on blood pressure, heart rate, and on some circulating vasoactive hormones, such as plasma renin activity, catecholamines, serotonin and endothelin 1 in ten healthy normotensive volunteers and eight moderate-severe essential hypertensives were studied. Two milligrams doxazosin was administered orally in randomised fashion as compared to placebo and the acute effects were evaluated one half to four hours later. Doxazosin did not reduce blood pressure and did not increase heart rate in normotensive subjects as compared to placebo. A significant decrease in blood pressure was found in hypertensives after doxazosin (p < 0.01), without change in heart rate. Simultaneously, doxazosin did not modify plasma renin activity, catecholamines, serotonin and endothelin-1 concentrations both in normotensive and hypertensive subjects when compared with placebo administration. Thus, it appears that doxazosin does not influence local and systemic vasopressor hormones involving in the regulation of blood pressure. In conclusion, the lack of effects on plasma vasoactive factors confirms the selective alpha-1 postsynaptic antagonism of doxazosin and an effective antihypertensive action in the treatment of essential hypertension. PMID- 7480966 TI - A new dry chemistry immunoassay: comparison with a fluorescence polarization system. AB - A new dry chemistry fluorescence immunoassay has been recently developed for therapeutic drug monitoring. In the present study we compared this new assay with a fluorescence polarization immunoassay. The drugs tested were carbamazepine, phenobarbital, phenytoin, theophylline and valproate. An high degree of correlation (coefficients ranging from 0.95 to 1.00) was obtained for all drugs tested. PMID- 7480967 TI - [Glucose homeostasis and the counterregulation system, hypoglycemia in transitory CRF deficit]. PMID- 7480968 TI - [Acute manifestations of diabetic gastroparesis]. PMID- 7480969 TI - Treatment of myocardial infarction: hypotensive effect of different thrombolytic agents. AB - The incidence of hypotension in patients treated with thrombolytic agents for myocardial infarction was investigated in a series of 71 patients, 17 treated with urokinase, 35 with rtPA and 19 with APSAC. Hypotension was observed in 23.5% of the first group, in 5.5% of the second, and in 42.10% of the third (p < 0.002 between rtPA and APSAC). In the inferior location hypotensive reaction was much more frequent than in anterior one (p < 007) especially if a right ventricular involvement was associated. Even if hypotension is a minor and generally harmless complication, it poses many practical problems, and its occurrence must be taken into account when choosing a fibrinolytic treatment. PMID- 7480970 TI - Lp(a) levels and reduced risk of vascular atheromatosis in patients with alcoholic liver disease. AB - In a group of 80 patients (50 male and 30 female) aged between 44 and 65, affected by alcoholic liver disease (46 with steatosis and 34 with liver cirrhosis) the Authors examined the relationship between the plasma lipid, in particular of Lp(a), and the incidence of vascular atherosclerotic plaques. The results were compared with those found in the controls (50 subjects of similar age, social and working status to that of the above patients but nondrinkers without liver or other metabolic disease). In the patients with steatosis we found a moderate increase in plasma lipid fractions including total, HDL and LDL cholesterol, but low levels of Lp(a), with an incidence of arterial plaques of 10.86%. In those with liver cirrhosis the findings were characterized by low levels of lipids and in particular of Lp(a), with an incidence of arterial plaques of 8.82%, decidedly less marked than in the controls (16%). In both cases the low incidence of vascular involvement appears to be in some way linked with low levels of Lp(a) and the severity of liver disease and not with the behaviour of HDL cholesterol. PMID- 7480971 TI - [Free radicals in anesthesia and the role of exogenous antioxidants]. AB - The alterations of the normal biological oxidoreductive balance can be due both to an increase of the plasmatic concentration of the free radical and the loss of the protective mechanisms. These conditions lead to the damages of cellular structure by the mechanism of lipoperoxidation. We have studied the lipoperoxidative effects of anesthetic drugs and the possible protective effect of the taurine. Two groups of 30 patients subjected to general anesthesia for a time of about 160 minutes, have been studied. The taurine has been administered in the second group, intravenously, before and during anesthesia. Blood samples were drawed in all patients in order to determinate plasmatic Malondialdehyde (MDA) and Glutathione peroxidase activities both in erythrocyte lysate and plasma. Our results revealed an increase of values of MDA and Glutathione peroxidase activities in the first group (without taurine). We think that this increase has been caused by the anesthetic drugs administration and that a protective cellular effect is developed by the taurine administration. PMID- 7480973 TI - Does it exist also a "late" post-surgical hypoparathyroidism? AB - Hypoparathyroidism following neck surgery, mainly thyroidectomy, is not a rare event, well known since many years. Post-surgical hypoparathyroidism may occur in form of two clinical syndromes of different etiology and prognosis. The first disease is a transitory hypoparathyroidism that might spontaneously recover within a few weeks or months. The second disease is a permanent hypoparathyroidism needing a definitive opoterapic treatment. Anyhow in both cases, hypocalcemic symptoms begin always within a short time from surgery, usually after an asymptomatic period elapsing from days to months. Only few cases of hypoparathyroidism clinically conclamate after many years from surgery have been reported. Description of a patient with hypoparathyroidism that became clinically evident thirty years after the thyroid surgery is herewith described. Our findings and review of a few cases reported by medical literature, can suggest a third form of post-surgical hypoparathyroidism with the distinctive feature of a very late beginning, probably following a long period of a latent parathyroid insufficiency. PMID- 7480974 TI - The importance of inflammation in early asthma. PMID- 7480972 TI - Helicobacter pylori in Barrett's esophagus and in normal or inflamed esophageal mucosa: a retrospective study. AB - To investigate on the presence and significance of Helicobacter pylori (Hp) in Barrett's mucosa, esophageal biopsy specimens from 34 patients with columnar lined epithelium were retrospectively examined. The incidence of colonization was compared with that observed in a control group of 76 subjects with normal esophageal mucosa or moderate esophagitis. Three of the 34 Barrett's specimens were Hp-positive and only 1 of these showed evidence of inflamed mucosa (p = NS). We do not find in this study any evidence to support the existence of association between Hp and Barrett's esophagus or esophageal inflammation. The presence of Hp in antral mucosa of all positive cases suggest the possibility that esophageal colonisation might merely be a consequence of reflux from the gastric antrum. PMID- 7480975 TI - The chemotherapy of tuberculosis--from the past to the future. PMID- 7480976 TI - Long-term oxygen therapy may improve skeletal muscle metabolism in advanced chronic obstructive pulmonary disease patients with chronic hypoxaemia. AB - Skeletal muscle metabolite depletion exists in advanced chronic obstructive pulmonary disease (COPD) patients with chronic hypoxaemia. The purpose of this study was to investigate if long-term oxygen therapy (LTOT) can improve skeletal muscle energy metabolism. Eight patients with advanced COPD, four with chronic hypoxaemia, were investigated using muscle biopsy specimens from the quadriceps femoris muscle applying the needle biopsy technique. The investigation was performed twice, before and after approximately 8 months of LTOT in the hypoxaemic patients. Eight healthy controls of similar age were also investigated. In the COPD patients, muscle glycogen, ATP and creatine phosphate (CrP) concentrations, were 42% (P < 0.01), 18% (P < 0.05) and 21% (P = n.s.) lower than in the healthy controls, respectively, while creatine (Cr) and lactate concentrations were 21% and 90% higher, respectively in the COPD patients compared to the healthy control subjects (P < 0.05). After LTOT, the 'energy index' CrP/(CrP + Cr) ratio increased by 0.12 in the LTOT patients but decreased by 0.12 in the control COPD patients (P < 0.05). The results indicate an improvement in skeletal muscle energy metabolism during LTOT in COPD patients with chronic hypoxaemia. PMID- 7480977 TI - Perception of dyspnoea during acute changes in lung function in patients with either asthma or COPD. AB - The aim of the study was to evaluate the relationship between several lung function indices and perceived dyspnoea during bronchoconstriction. Acute changes in lung function were induced by inhaled histamine followed by terbutaline, in 12 asthmatics and 12 subjects with chronic obstructive pulmonary disease (COPD). A bipolar visual analogue scale (VAS), allowing subjects to report either improvement or worsening when moving off from a 'nochange' midpoint, was used to rate shortness of breath. Large swings in ratings were seen in all asthmatics and in seven out of 12 COPD subjects (high perceivers). Using linear regression of VAS rating against parallel change in lung function, on a within-subject basis, the highest degree of correlation between dyspnoea and objective response was found to involve the change in specific inspiratory resistance (sRin) in the asthmatics. In the five low perceivers, the ability to discriminate an increase in airway obstruction, estimated as the VAS/change in lung function slope, was very poor. Using a stepwise multiple regression analysis, the sensation of dyspnoea was found to be significantly related to the FEV1 and the sRin in the asthmatics, to the inspiratory vital capacity and the maximal inspiratory flow at 50% FVC (MIF50) in the COPD subjects with high perception, and to the MIF50 in the COPD subjects with low perception. PMID- 7480978 TI - Assessment of the continued supervision and asthma management knowledge of patients possessing home nebulizers. AB - This paper describes the results of a questionnaire survey on some aspects of nebulizer use, nebulizer instruction, and features relating to the recognition and management of deteriorating asthma. The study population consisted of 90 children with home nebulizers attending a paediatric asthma clinic, and 200 asthmatic children and 200 asthmatic adults whose nebulizers had been purchased directly from a manufacturer in the U.K. The results suggest that follow-up supervision of the patients who bought their own nebulizers occurred in only approximately 25% of cases. Written information focusing on the management of symptoms was also lacking. Peak flow meters were being under-used. Although the majority (77-100%) of patients were aware of the '4 h rule' for repeat use of bronchodilator therapy at home, there was still some confusion about the acceptable time interval and action to be taken should the dose be required more frequently. Very few (32%) in the nationwide adult group had a crisis action plan should the nebulizer fail to produce relief. This paper recommends that a simple treatment and crisis plan should be included with the purchased nebulizer, highlighting the key areas to be discussed by the patients with the doctor or other medical professionals. Such a package may prevent the occasional disaster that can occur with unsupervised nebulized bronchodilator usage. PMID- 7480979 TI - Why disease due to Mycobacterium tuberculosis is less common than expected in HIV positive patients in Edinburgh. AB - By December 1993, only five cases of tuberculosis were observed in the 1030 HIV positive patients in Edinburgh, U.K., although, on the basis of historical tuberculin skin test data, between four and eight new cases of tuberculosis were expected per year. Of 310 HIV-positive patients, none of the 19 (6.1%) who were tuberculin skin test positive had developed tuberculosis after 87 months (average) of follow-up. It is suggested that new or re-infection is a more common cause of tuberculosis in HIV-positive patients than reactivation. Restriction fragment length polymorphism typing of Mycobacterium tuberculosis strains could confirm this hypothesis and support currently suggested additional infection control procedures. PMID- 7480980 TI - Two years experience with recombinant human DNase I in the treatment of pulmonary disease in cystic fibrosis. AB - Recombinant human DNase I (rhDNase) has been shown to improve pulmonary function in patients treated for up to 6 months. A cohort of 52 cystic fibrosis patients with a FVC > 40% predicted were enrolled into an open label study in order to evaluate longer-term effects of rhDNase. They received 2.5 mg rhDNase twice daily for 6 months followed by a 2-week wash-out period, and for the subsequent 18 months were treated with rhDNase once daily. Twenty-six male and 26 female patients with a mean FVC of 2.941 and FEV1 of 1.471 were recruited. Thirteen patients did not complete the study; there were seven deaths, three patients withdrew consent and three patients were lost to follow-up. Improvement in pulmonary function was seen following treatment and changes were evaluated as mean percent change from baseline. The maximum improvement occurred in the first month followed by a plateau at a lower level of improvement. The mean improvement in FEV1 over the first month was 13.3% (range 12-14.1%), followed by a plateau at around 7.1% (range 4.6-11.0%) for the subsequent 23 months. Mean FVC was improved by 12.03% (range 9.0-14.3%) over the first month and subsequently 4.2% (range - 2.2-10.2%). The effects on pulmonary function were similar for both treatment doses of rhDNase. There was also a steady improvement in weight from a mean of 54.2 kg to 55.7 kg at the end of the study.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7480981 TI - Effect of menthol vapour on airway hyperresponsiveness in patients with mild asthma. PMID- 7480982 TI - Distribution of human neutrophil elastase in diffuse alveolar damage and pneumonia in a case of neonatal sepsis. PMID- 7480983 TI - Pleural effusion in ankylosing spondylitis; successful treatment with intra pleural steroid administration. PMID- 7480984 TI - Assessing equivalence of inhaled drugs. AB - The move to disease management has led to an increase in the practice of drug or formulation substitution on the basis of equivalence. Well established guidelines are available for judging equivalence between oral, but not inhaled, formulations. This article describes the criteria by which equivalence can be assessed and concludes that although traditional issues such as adequate sample size are important, studies also need to be designed in such a way as to avoid the possibility of falsely concluding clinical equivalence. PMID- 7480985 TI - Measurement of respiratory muscle strength. PMID- 7480986 TI - Comparison of four-times-a-day and twice-a-day dosing regimens in subjects requiring 1200 micrograms or less of budesonide to control mild to moderate asthma. AB - The aim of this study was to compare the efficacy, compliance and side-effects of budesonide administered twice daily (b.d.) and four times a day (q.d.) with a Turbuhaler device in asthmatic subjects requiring < or = 1200 micrograms daily. The randomized, parallel group study design included a 2-week baseline period followed by a 6-12-month treatment period. Subjects were assessed at regular intervals in hospital through FEV1, PC20 methacholine, adrenal function and throat swabs. They were asked to record their symptoms and PEF values morning and evening at home. An asthmatic flare-up, which was the main outcome resulting in a patient's termination of the study, was defined beforehand as (a) 25% or greater diurnal variability in PEF for 2 consecutive days, and/or (b) nocturnal awakenings due to asthma symptoms 2 days or more in the same week and/or (c) an increase (doubling or more) in the need for inhaled bronchodilator 2 days in the same week. Fifty-eight adult asthmatic subjects (20 males and 38 females) entered the study, one-half being randomly assigned to the b.d. regimen and one half to the q.d. regimen. Fourteen subjects were on 400 micrograms, 15 subjects on 800 micrograms and 29 subjects on 1200 micrograms of budesonide daily. Seventeen flare-ups were recorded in the b.d. regimen group as opposed to 11 in the q.d. regimen (P = 0.05), significant differences being found in the 800 and 1200 micrograms groups (a total of 13 flare-ups in the b.d. group and eight flare-ups in the q.d. group for the two doses, P = 0.01).(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7480987 TI - The role of fine needle aspiration in the diagnosis of palpable metastases from lung cancer. PMID- 7480989 TI - Miliary tuberculosis in a high prevalence area of the U.K.: Blackburn 1978-1993. AB - Thirty-nine cases of military tuberculosis treated between 1978-1993 in a district with a high incidence of tuberculosis are reported. Twenty-eight patients were of Indian Subcontinent (ISC) ethnic origin, 10 patients were Caucasian and one patient was Chinese. Mortality at 10% in both Caucasian (chi2 4.94; 0.05 > P > 0.01) and ISC patients (chi2 10.22; 0.001 > P) were significantly lower than in a recently reported series from Edinburgh, as was the proportion of post mortem diagnoses of miliary tuberculosis (chi2 6.35; 0.02 > P > 0.01). Both the rate of miliary tuberculosis and the average age at diagnosis in Caucasian patients were lower than in the Edinburgh series suggesting that miliary tuberculosis could be under-diagnosed in the elderly Caucasian population in the Blackburn district. The better outcome of those diagnosed in life may be partly due to heightened awareness of tuberculosis locally and partly due to treatment by a centralized tuberculosis service. PMID- 7480988 TI - Extrinsic and intrinsic asthma from childhood to adult age: a 10-yr follow-up. AB - BACKGROUND: Asthma constitutes one of the most common chronic diseases in childhood, yet little is known about the factors that determine the outcome in childhood asthma. The purpose of this study was to describe various factors of potential importance for the outcome in children with intrinsic and extrinsic asthma. METHODS: Of 85 consecutive children, 5-15 years of age with asthma, 70 (82%) participated in a 10-yr follow-up examination. At the time of referral, all children underwent certain tests for asthma (case history, total IgE, skin prick tests, radioallergosorbent (RAST) tests and specific bronchial provocations). On the basis of these tests, 24 children had intrinsic asthma and 46 children had extrinsic asthma. RESULTS: At the follow-up examination, 60 of the 70 adults (86%, 20 with intrinsic asthma) had current symptoms; 54 of the 60(90%; 18 with intrinsic asthma) were receiving maintenance therapy. Both FEV1 %predicted and FEV1/FVC increased significantly (from 73% +/- 19% to 92% +/- 17% and from 75% +/ 13% to 80% +/- 12% respectively) from childhood to early adulthood (P < 0.0001 and P < 0.001), respectively); a matching amelioration of symptoms was observed (P < 0.0001). Adults with current symptoms had a significantly lower FEV1 %predicted at the time of follow-up (90% +/- 2% vs. 100% +/- 4%, P < 0.02) but not in childhood (73% +/- 20% vs. 71% +/- 10%) than adults who had ceased wheezing. CONCLUSIONS: In children with intrinsic asthma, the outcome seems to be predicted by a combination of the initial frequency of symptoms (P = 0.04), initial FEV1 (P = 0.002), active smoking (P = 0.001) and age at onset of respiratory symptoms (P = 0.001), whereas the initial FEV1 (P < 0.001) seems to be a strong predictor for the outcome in children with extrinsic asthma. These findings suggest that the pathogenic mechanisms underlying intrinsic and extrinsic asthma in children may differ. PMID- 7480990 TI - Influenza immunization status and viral respiratory tract infections in patients with chronic airflow limitation. PMID- 7480991 TI - Thoracoscopic bullectomy and tetracycline pleurodesis for the treatment of spontaneous pneumothorax. AB - Initial experience of thoracoscopic bullectomy and tetracycline pleurodesis for the treatment of spontaneous pneumothorax is reported. Thirty-three out of 49 patients admitted with spontaneous pneumothorax were suitable for treatment with this minimally invasive method. This series demonstrates that this surgical management offers early discharge and return to normal activities with excellent medium-term results, despite the three early failures. It is felt that with increased experience in thoracoscopy and improved selection of patients, thoracoscopic bullectomy and pleurodesis will become the treatment of choice for primary spontaneous pneumothorax. PMID- 7480992 TI - A drying chamber for use with small volume jet nebulizers. PMID- 7480993 TI - Bedside fiberoptic bronchoscopy for tracheostomy decannulation. PMID- 7480994 TI - A case of chronic gastrointestinal blood loss in cystic fibrosis. PMID- 7480995 TI - Paediatric endobronchial leiomyoma mimicking asthma. PMID- 7480996 TI - Audit of procedures related to outcome of chest clinic consultation. PMID- 7480999 TI - [French Society of Orthopedic Surgery and Traumatology. Membership list]. PMID- 7480998 TI - Nocturnal hypoxaemia in cystic fibrosis. PMID- 7480997 TI - Recurrent laryngeal palsy and mediastinal lymphadenopathy. PMID- 7481000 TI - [Vaccination of the elderly]. PMID- 7481001 TI - [Whipple's disease: more on the identity of its etiological agent]. PMID- 7481002 TI - [Autosomal dominant polycystic kidney disease. A comparative genetic and clinico radiological study between patients with normal renal function and their healthy relatives]. AB - We studied 6 families with autosomal dominant polycystic kidney disease (ADPKD), to compare patients with normal renal function and healthy family members. From 38 subjects, 20 were carried of the genetic abnormality associated to the disease (however, in 4 renal cysts were absent) and 18 were healthy relatives. We did not found any difference between the two groups in respect to clinical manifestations, physical examination or blood chemistry and renal function tests studied, except an increase of phosphaturia in ADPKD patients. However, an increase in kidney size in ADPKD was noted, more evident in hypertensive patients. The blood pressure was correlated with kidney size in patients with ADPKD, but not in healthy relatives. We conclude that in early stages of the disease, patients with ADPKD do not have clinical or analytical abnormalities although they have an increase in kidney size. Genetic studies have great value to detect patients in early stages of the disease. PMID- 7481003 TI - [The clinical efficacy and complications of the transjugular intrahepatic portosystemic shunt-stent]. AB - TIPSS is a new therapeutic modality for decompressing the portal tree and its use has broadened in the last five years. From February 1993 to August 1994 a prospective study was performed to evaluate its efficacy and safety. Nineteen cirrhotic patients (Child A-5, B-10, and C-4) with a TIPSS placed were included. The mean follow-up was 7.2 months. The indication was therapy of esophageal variceal bleeding in 18 patients (acutely in 8 and elective in 10 patients) and refractory ascites in one. In all cases could the "stent" be placed and the portocaval gradient decreased from 22.8 +/- 3.71 to 9.3 +/- 2.27 mmHg. In the first thirty days the mortality rate was 10.5%, with the following complications: two portal thromboses, two acute non-lithiasic cholecystitis, one hemoperitoneum, one spontaneous bacterial peritonitis, and one hepatic encephalopathy. During the follow-up period two patients developed hemorrhagic relapses and two additional patients subclinical encephalopathy. TIPSS dysfunction was observed in 57.8%. PMID- 7481004 TI - [Acute pancreatitis in patients infected with the human immunodeficiency virus]. AB - This study was designed to define the etiological spectrum of acute pancreatitis in patients infected with HIV in our environment and to evaluate the relevance of cytomegalovirus as etiological agent. A retrospective analysis was first made of clinical, analytical, radiological and pathological findings of patients infected with HIV clinically diagnosed of acute pancreatitis (period 1984-1993). Then, clinical records of patients with necropsy proven CMV pancreatic involvement were retrospectively analyzed (period 1985-1992) to evaluate whether they had been diagnosed of acute pancreatitis when alive. Nine cases were diagnosed of acute pancreatitis. A presumptive etiology (didanosine) was considered in seven cases and idiopathic in two. Five patients died, two from causes directly related to acute pancreatitis. Necropsy was performed in two cases. Mortality was associated with the development of renal insufficiency or the appearance of three or more complications (p = 0.039). With regard to the second phase of the study CMV pancreatic involvement was demonstrated in 4 necropsies out of a total of 22 with evidence of CMV infection in one or more organs. None of these patients was diagnosed of acute pancreatitis when alive. Acute pancreatitis is an uncommon entity in patients infected with HIV. Presumptively, it could be attributed to didanosine therapy in most of our cases. The mortality rate was high, particularly when renal failure developed. In no case was CMV involved as etiologic agent when the patient was alive, although necropsy showed that pancreas can be subclinically involved in the context of a CMV disseminated infection. PMID- 7481005 TI - [Toxic myopathies related to the administration of hypolipidemic agents: are the drugs the only things responsible?]. AB - The characteristics are reported of five myopathies initially considered toxic in nature caused by the administration of blood lipid lowering agents. In three cases the final diagnosis was hypothyroidism, thus demonstrating the need to rule out the potential causes of secondary dyslipidemia before initiating therapy. Likewise, and because of the rare true muscle involvement from blood lipid lowering agents, it is necessary to investigate those diseases or factors enhancing muscle toxicity when it appears. PMID- 7481007 TI - [Whipple's disease: an entity to keep in mind]. PMID- 7481006 TI - [Poisoning by organophosphates and paraquat: apropos 14 cases]. AB - We present the clinical aspects and the evolution of 14 patients brought in the last 5 years in an ICU polyvalent with diagnostic of intoxication by organophosphate (OF) 10 and paraquat 4. The intoxicated by OF were subjugated at the same treatment record book with atropine, gastric washing, coal activated and pralidoxime. At the intoxicated by paraquat used precocious hemoperfusion. The mortality in intoxicated by OF went 20%, and 100% in paraquat intoxications. We analyzed to them the complications during their stay in ICU, as soon as diverse clinical aspects of this intoxications and equal form varied aspects about treatment. PMID- 7481009 TI - [The pathogenetic mechanisms of dyspnea]. PMID- 7481008 TI - [Cystic fibrosis in adults: the new approach to an old disease]. PMID- 7481010 TI - [The role of emergency therapeutic endoscopic retrograde cholangiopancreatography in acute biliary pancreatitis]. PMID- 7481011 TI - [Pulmonary infiltrates and respiratory failure in a heart transplant patient with a primary cytomegalovirus infection]. PMID- 7481012 TI - [Diarrhea, a constitutional syndrome and arthralgias]. PMID- 7481013 TI - [Cavitary pneumonia in a neutropenic patient]. PMID- 7481014 TI - [A solitary pulmonary nodule in an alcoholic patient]. PMID- 7481015 TI - [Hemiparesis-ataxia in an HIV-positive man]. PMID- 7481016 TI - [Arterial hypertension does not constitute the principal cerebrovascular risk factor in different subtypes of stroke]. PMID- 7481017 TI - [Acute brucellosis and rhabdomyolysis]. PMID- 7481018 TI - [New momentum for the journal]. PMID- 7481019 TI - [Blood saving]. PMID- 7481020 TI - [Propofol in continuous perfusion as anesthetic in experimental surgery in the rabbit]. AB - OBJECTIVES: To study anesthesia with intramuscular ketamine chlorohydrate and continuous intravenous perfusion of propofol (2.6 phenyl diisopropyl) in 30 white New Zealand rabbits undergoing tracheal and vascular surgery and experimental bronchoscopy without mechanical ventilation. MATERIAL AND METHOD: The animals were monitored by transcutaneous pulse oximetry and were sedated before canalization of the auricular vein. Oxygen was administered under a bell at a rate of 5 l/min. Variables recorded were heart and respiratory rates, oxygen saturation, blink reflex, and movements during surgery. The anesthetic protocol consisted of sedation (intramuscular ketamine chlorohydrate 50 mg/kg), followed by induction (intravenous propofol 3 mg/kg), and maintenance with intravenous infusion of diluted propofol in 2 phases as follows: a loading dose of 1.2 mg/kg/min first, followed by maintenance with 0.9 mg/kg/min propofol in dextrose at 5% concentration (4 mg/ml). RESULTS: The procedure afforded adequate anesthesia for surgery lasting up to 70 min. Vital signs were stable in all animals. The perfusion rate was modified during surgery in 8 animals, in 5 because of superficial anesthesia and in 3 because of respiratory depression. Respiratory rate was the only parameter that was significantly correlated with depth of anesthesia and propofol dose. Postanesthetic recovery was rapid in all animals. No cases of heart/respiratory failure or death occurred. CONCLUSIONS: The proposed method is useful for anesthetizing the rabbit, as it renders mechanical ventilation unnecessary. Only simple monitoring of respiratory rate is required. PMID- 7481021 TI - [Infusion of propofol as anesthetic and antiemetic drug]. AB - OBJECTIVES: To determine the possible antiemetic effect of propofol as the single agent for intravenous anesthesia in children undergoing strabismus surgery, in comparison with other techniques. PATIENTS AND METHOD: Eighty ASA I and II patients between 3 and 10 years, divided into 4 groups of 20, were studied prospectively. Groups I, II and III were premedicated with dehydrobenzylperidol (90 micrograms/kg) i.m. Group I then received sleep doses of thiopental and N2O. Group II received propofol, sleep doses, and N2O. Groups III and IV received sleep doses of propofol followed by intravenous perfusion (10 mg/kg/h). RESULTS: Half the patients in group I experienced postoperative vomiting, more than in groups II, III and IV (p < 0.05). The incidence of vomiting in group II was 30%. Groups III and IV experienced less vomiting (5-10%) than did groups I and II (p < 0.05). The incidence of bradycardia during surgery was higher in groups III and IV (p < 0.05); medical treatment was not required. CONCLUSIONS: Propofol used as the single agent for induction and maintenance of anesthesia offers clear advantages in reducing emesis after strabismus surgery. PMID- 7481023 TI - [Comparative study of continuous epidural analgesia versus intermittent, patient controlled, epidural analgesia during labor]. AB - OBJECTIVE: To assess the analgesic efficacy and the incidence of side effects of three procedures of epidural analgesia during labor. PATIENTS AND METHODS: One hundred and one nulliparous pregnant women at term with cephalic presentation were randomly allocated to 3 groups (A: 31, B: 35, C: 35). Group A was given an initial dose of 8 ml bupivacaine 0.25% (20 mg) with epinephrine 1/200,000 and 50 micrograms fentanyl followed by a continuous infusion of 8 ml/h bupivacaine 0.125% (10 mg/h) with epinephrine 1/400,000. The initial dose in group B was 10 ml bupivacaine 0.125% (12.5 mg) with epinephrine 1/400,000 and 50 micrograms fentanyl followed by a continuous infusion of 12 ml/h bupivacaine 0.0625% (7.5 mg/h) with epinephrine 1/800,000 and 12 micrograms/h fentanyl. In group C the initial dose was the same as for group A, but maintenance was provided through a patient-controlled (PCA) system with 5 ml bupivacaine 0.25% (12.5 mg) and epinephrine 1/200,000 set for a minimum interval of 1 h. If analgesia was inadequate all patients could receive top-ups of 5 ml bupivacaine 0.25% (12.5 mg) with epinephrine 1/200,000. RESULTS: Quality of analgesia was similar in all three groups. We found no significant differences in level of sensory/motor blockade, hemodynamic changes, side effects or type of delivery. The total dose of bupivacaine was highest in group A, whereas group B received the higher total dose of fentanyl. The number of Apgar scores under 8 was highest in group A, although the scores rose within 5 min of delivery in all cases. CONCLUSIONS: We believe that the protocol of choice is the one used with group B (initial dose of 10 ml bupivacaine 0.125% + epinephrine 1/400,000 + 50 micrograms fentanyl followed by 12 ml/h bupivacaine 0.0625% + epinephrine 1/800,000 + 12 mu cg/h fentanyl in continuous infusion). The use of PCA systems with our unscreened patients yielded no advantages over the procedure used with group B. PMID- 7481025 TI - [Physiopathologic implications of the anesthesiologic management of pediatric laparoscopic surgery]. AB - We study gas exchange and hemodynamic repercussions during pediatric laparoscopic surgery. We provided balanced anesthesia with muscle relaxation while ventilation was maintained with FiO2 at 0.4 and flow volume between 10-15 ml/kg-1 during abdominal laparoscopic procedures performed in 10 ASA I-II children (4-14 years). Pneumoperitoneum was produced with CO2 insufflated up to a pressure of 15 mmHg. Airways pressure (PIP), PaO2, PaCO2, heart rate (HR), systolic arterial pressure (SAP) and diastolic arterial pressure (DAP) were measured before insufflation and 5, 30 and 60 minutes afterwards. We calculated the ratio of dead space to flow volume (VD/VT), thoracic distensibility and metabolic production of CO2 (VCO2). Insufflation caused an immediate reduction (29-33%) in dynamic thoracic distensibility (p = 0.0004), but no hypoventilation or increases in VD/VT. The decrease in PaO2 was small (5-6%) but statistically significant (p = 0.0188). Hypercapnia (14-21%) was due to an increase in VCO2 caused by gradual peritoneal absorption of CO2 (24-32%, p = 0.0013). We also found increases in SAP (10%, p = 0.02) and DAP (32%, p = 0.0001) at 5 min, along with an increase in HR (8%, p = 0.0163) at 60 min. Arterial CO2 levels were held within physiological limits by compensatory hyperventilation (+25% of physiological VT). Capnography proved to be an excellent guide. Any clinical repercussion of hemodynamic effects was blocked by a dose of atropine given before insufflation and by the excess loading volume (10 ml/kg of crystalloids). Laparascopic surgery in children diminishes thoracic distensibility and causes hypercapnia, making it necessary to measure PefCO2 to regulate ventilation. PMID- 7481024 TI - [A case of malignant hyperthermia: anesthesia and recovery]. AB - We report the case of a 29-years-old woman with Meniere's syndrome undergoing surgery to drain the endolymphatic sack. After anesthetic induction trismus developed but tubes were inserted without difficulty. Tachycardia began 15 min after start of balanced anesthesia with isoflurane, as arterial oxygen saturation and CO2 pressure decreased and temperature increased. These symptoms responded favorably to immediate treatment with dantrolene sodium, which was continued for 30 h in the ward for postoperative intensive care. Blood chemistry reports showed high enzyme levels that peaked at 24 h. We stress the importance of monitoring end tidal CO2 pressure and of rapidly starting specific treatment with dantrolene for correction of the clinical picture described. PMID- 7481022 TI - [Isoflurane versus propofol for sedation after heart surgery]. AB - OBJECTIVE: To compare isoflurane and propofol as the principal sedatives in ventilated patients after cardiac surgery with extracorporeal circulation (ECC). PATIENTS AND METHODS: Forty consecutive, randomized patients undergoing cardiac surgery with ECC were studied prospectively. Patients were assigned to receive either isoflurane or propofol. Variables recorded were level of sedation, adaptation to the respirator and hemodynamic stability. RESULTS: Each group included 20 patients. The mean dose of isoflurane was 0.39% +/- 0.06% of end tidal concentration. The mean propofol dose was 2.11 +/- 0.55 mg/kg/h. Statistically significant differences were found for stabilization of sedation time (4 min +/- 1.17 for isoflurane and 11.7 min +/- 4.78 for propofol) and time to endotracheal extubation (56.2 min +/- 20.47 for isoflurane and 72.65 min +/- 30.90 for propofol), number of times dosage had to be changed (2.20 +/- 0.89 with isoflurane and 7.05 +/- 2.58 with propofol) and time of administration had to be interrupted (8.45 min +/- 8.73 with propofol and 0.75 min +/- 1.94 with isoflurane). CONCLUSIONS: In comparison with propofol, isoflurane offers advantages of rapid stabilization of sedation, ease of use and shorter time to endotracheal extubation. PMID- 7481026 TI - [Anesthesia with continuous + infusion of propofol for trans-sternal thymectomy in myasthenic patients]. AB - Patients with myasthenia gravis respond unpredictably to muscle relaxants and more often suffer respiratory complications after surgery. We describe the use of total intravenous anesthesia with propofol and alfentanil without muscle relaxants is three myasthenic patients classified as Osserman I-IIB. Mean time since appearance of the disease was 1 year and all were undergoing transsternal thymectomy. Time in surgery ranged from 115 to 170 min and mean total dose of propofol was 1,374 mg. In all cases total intravenous anesthesia afforded good conditions for intubation, maintenance during surgery and rapid recovery from anesthesia, with early extubation. PMID- 7481027 TI - [Analysis of preoperative reserve criteria and intraoperative transfusion of blood derivatives at an anesthesiology and recovery service]. AB - Three hundred and three patients undergoing elective surgery were studied prospectively. Patient data and results of serial blood cell counts were recorded before and after surgery and used to evaluate the criteria used to reserve and transfuse blood and blood products, as well as to assess the outcome of transfusions performed. Six hundred and ninety-nine units were reserved, indicating a reservation-to-transfusion ratio of 1.8. The most often cited reasons for reserving blood were surgical technique, nature of disease and prolonged surgery. A high number of reservations were called for from the ward by the surgeons themselves. A total of 180 patients were given transfusions, the main reason being the anesthesiologist's judgement that the Hb count was low (in 26% of cases). Transfusions because of slight bleeding during surgery and prolonged surgery gave rise to the highest Hb counts. The lowest post-transfusion Hb counts recorded occurred when the reason cited was presence of signs of anemia. We conclude that the ratio of reservations to transfusions in our study was acceptable, as were the reasons justifying reserves. There is an overall tendency to achieve excessively high Hb counts (10-11 g/dl) in the early postoperative period. The best criterion for performing transfusion is the presence of signs of anemia. PMID- 7481028 TI - [Methods of blood saving in the surgical patient]. PMID- 7481029 TI - [Treatment of reflex sympathetic dystrophy with continuous blockade of the brachial plexus]. PMID- 7481030 TI - [Sedation of children for CAT: assessment of chloral hydrate]. PMID- 7481031 TI - [Remembering Juan Vicente Hedo's ether (1847). Chronicle of an unexplained oversight]. PMID- 7481033 TI - [An analysis of heart rate variability. Spectrum analysis methods and complex signal demodulation]. AB - The analysis of heart rate variability provides valuable information in the noninvasive study of neurovegetative activity and its modifications induced by drugs, physiological maneuvers or disease processes, and in the evaluation of prognosis and risk stratification in different cardiovascular diseases. A review is made of the different methods used to study heart rate variability, and an account is given of the information provided by spectral methods and the new procedures based on the complex demodulation of the time series composed of consecutive cardiac cycles. The limitations of the spectral methods are deal with, particularly in defining the time-dependent changes in variability and their relation to clinical events. Likewise, a description is given of the ability of complex demodulation to define the time course of the oscillations into which the analyzed time series is decomposed. Complex demodulation based on the fast Fourier transform and its inverse is able to establish the instantaneous amplitude and frequency of each of the oscillations contained in the time series, separated by specific filters in the previously selected frequency bands (high: 0.15-0.40 Hz, low: 0.04-0.15 Hz, or very low: < 0.04 Hz), and from which the original signal may be reconstructed. The evaluation of the different methodological approaches, and the analysis of the causal relations between the variability modifications and clinical events will further extend the clinical relevance of the study of heart rate variability. PMID- 7481032 TI - [Rehabilitation of the coronary patient. Secondary prevention. A document of the Working Group on Cardiac Rehabilitation of the Sociedad Espanola de Cardiologia]. AB - There is scientific evidence that these measures of secondary prevention and cardiac rehabilitation, not only diminishes the mortality and morbidity of the patients but also favour the return to work, reduce incapacities and improve the quality of life. The economic cost of these methods must not be an obstacle for the Administration, as recent studies about the cost-benefit analysis of these programmes have demonstrated. PMID- 7481034 TI - [Acute myocarditis with severe cardiac dysfunction in the pediatric population. The evolution and differential characteristics with respect to adult myocarditis]. AB - AIMS: The aim of our study was to assess the spontaneous outcome of acute myocarditis associated with severe cardiac dysfunction in children, as well as to compare these features with those occurring in adult patients. METHODS: Fifty patients consecutively diagnosed of acute myocarditis during the last 7 years in our hospital were studied; 15 patients were children younger than 14 years, and 35 were adults. Immunosuppressive therapy was not used in any patient. RESULTS: Mean age was 2 +/- 3 years in children, ranging from 2 months to 12 years. One patient required temporary pacing for a third-degree atrioventricular block, while the remaining 14 children had severe congestive heart failure, with a left ventricular ejection fraction of 30 +/- 12% (16 to 44%). After a mean follow-up of 21 +/- 26 months, only 3 children died, at 1, 4 and 10 months after the initial diagnosis. Death was sudden in all 3 patients. Left ventricular ejection fraction rose to 45 +/- 14% at 1 month after diagnosis, and to 58 +/- 15% at the end of follow-up. Unfavorable evolution (death or evolution to chronic dilated cardiomyopathy, with a left ventricular ejection fraction < 45%) occurred in 6 children (40%) at 1 month after diagnosis and in only 4 (25%) at the end of follow-up. The 9 children with 1-month favorable outcome were alive and had an ejection fraction > 45% at long-term, while only 2 of the 6 children with 1-month unfavorable outcome were alive and had an ejection fraction > 45% at long-term. Only the 3 children who died had an ejection fraction < 30% at 1-month. Favorable outcome was more frequent in children that in adult patients with acute myocarditis (75% versus 46%). CONCLUSIONS: The outcome of acute myocarditis with severe cardiac dysfunction was favorable in a majority of pediatric patients; this favorable evolution was less frequent in adults. Patients in whom left ventricular ejection fraction did not increase at short-term had a higher risk of death, and they should probably be considered for heart transplantation. PMID- 7481035 TI - [The incidence of major histocompatibility system antigens in dilated and ischemic myocardiopathies]. AB - AIM: The purpose of this study was to analyze the frequency of the different antigens of HLA in patients with diagnosis of very advanced dilated cardiomyopathy and ischemic heart disease by comparing them with a control group of supposedly healthy subjects. MATERIAL AND METHOD: The group of dilated cardiomyopathy consisted of 35 patients (8 women and 27 men) aged between 14 and 60 years. The group of ischemic heart disease included 32 patients (4 women and 28 men) aged between 34 and 64 years. The control group comprised 1337 subjects of the Spanish Mediterranean area, supposedly healthy and recruited from paternity studies. RESULTS: In dilated cardiomyopathy we found a higher incidence in comparison with the control group of the A-2 (62.86% vs 46.22%), B-12 (60.00% vs 32.38%) and DQ-3 (82.86 vs 49.96%) antigens, and a lower incidence of B-51 (0.00% vs 12.49%). In ischemic heart disease we found, when comparing to the control group, a higher incidence of A-11 (31.25% vs 13.08%) and A-29 (34.38% vs 14.58%) antigens and a lower incidence of DQ-2 (15.63% vs 49.88%). CONCLUSIONS: In the Spanish Mediterranean area, the presence of A-2, B-12 and DQ-3 antigens, as well as the absence of B-51 would favour the appearance of advanced dilated cardiomyopathy. The presence of the A-11 and A-29 antigens would predispose to ischemic cardiomyopathy while the presence of DQ-2 would have a protective effect on the appearance of this cardiopathy. PMID- 7481036 TI - [The usefulness of the isovolumetric relaxation time of both ventricles in detecting acute rejection in the heart transplant patient]. AB - AIMS: The purpose of our study was to evaluate the usefulness of the isovolumetric relaxation time in both ventricles when diagnosing acute rejection in transplanted patients. METHOD: 68 endomyocardial biopsies were performed on a total of 38 patients. An echocardiographic study was carried out within the first 24 hours of each biopsy. All registrations were made by the same person. The isovolumetric relaxation time was measured in the left and right ventricles. The patients were divided according to two criteria: according to the degree of rejection (0-I, II, III) and according to whether the rejection was treatable (III) or non-treatable (0, I and II). RESULTS: In both ventricles, there was a progressive decrease of the isovolumetric relaxation time corresponding to higher degrees of rejection: in the left ventricle (0-I = 90 +/- 16; II = 74 +/- 16; III = 70 +/- 26; significant differences of II and III in relation to 0-I) as well as in right ventricle (0-I = 43 +/- 16; II = 37 +/- 14; III = 29 +/- 8; significant difference of III in relation to 0-I). The patients with treatable and non treatable rejection were compared: no differences were found in the isovolumetric relaxation time of the left ventricle (0, I and II = 85 +/- 16 vs III = 70 +/- 26), but they were found in the right ventricle (0, I and II = 41 +/- 15 vs III = 29 +/- 8). CONCLUSIONS: Acute heart rejection induces a decrease of the isovolumetric relaxation time in both the left ventricle and the right ventricle. However, the isovolumetric relaxation time of the right ventricle seems to be a more useful parameter than isovolumetric relaxation time of the left ventricle, as it permits to detect whether an acute heart rejection is treatable or non treatable. PMID- 7481037 TI - [Myocardiopathies (II). Genetic changes in the etiopathogenesis of hypertrophic myocardiopathy. The therapeutic prospects]. AB - The genetical characterization of any disease implies its immediate theoretic and practical reorganization since all the basic clinical aspects such as ethology, diagnosis, prognosis, prevention and finally treatment are affected. This can be the fact for hypertrophic myocardiopathy in the near future. Recently, mutations in some new genes causing this alteration, apart from those found in the beta myosin heavy chain gene, have been identified. Hypertrophic cardiomyopathy could be classified etiopathogenetically as primary, if it is due to a genetic alteration in any of the components of the sarcomere, and secondary when the initial factor is external, although it will be eventually reflected in a malfunction of the sarcomere. Therefore hypertrophic cardiomyopathy could be defined as a myocardial disorder with and autosomic hereditary pattern which is characterized by a ventricular hypertrophy due to alterations in the cardiac sarcomere. Detected mutations so far, which have been admitted to be the primary alteration in this disease, are localized in the beta-myosin heavy chain gene (14q1), in the alpha-tropomyosin gene (15q2), in the cardiac troponin T gene (1q3), as well as in the chromosome 11 p13-q13 loci. Different authors have pointed out a possible epigenetic mechanism produced by endogenous or environmental secondary factors as responsible for hypertrophic myocardiopathy.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7481038 TI - [A paradoxical intraventricular gradient in a patient with atypical hypertrophic myocardiopathy]. AB - A paradoxic diastolic intraventricular flow (directed from apex to base), caused by a pressure gradient between a small apical aneurysm and the main left ventricular cavity, connected by a narrow muscular tunnel, was detected by Doppler echocardiography in a 69 year old male with apical hypertrophic cardiomyopathy and effort angina. Coronary arteriography was normal and the clinical, electrocardiographic, echocardiographic and angiocardiographic features of the patient were otherwise typical of apical hypertrophic cardiomyopathy. Paradoxic diastolic gradients have been occasionally described in patients with hypertrophic cardiomyopathy, and indicate the existence of a sequestered apical cavity, which is frequently not detected by echocardiography. Identification of these gradients may have important clinical implications, since they have been related with a higher incidence of embolic events, severe ventricular arrhythmias and myocardial ischaemia and necrosis. PMID- 7481039 TI - [A retroaortic anomalous left coronary artery]. AB - We report a 67-years-old patient with angina pectoris and positive treadmill test. In the coronary arteriography the left coronary artery arose from the right coronary sinus of Valsalva through an independent ostium and followed a retroaortic course. Coronary artery disease was no demonstrated, so we thought that the myocardial ischaemia shown in the treadmill test, was due to the anomalous origin of the left coronary artery. Surgical treatment was recommended. PMID- 7481040 TI - [Bilateral pulmonary thromboendarterectomy in chronic pulmonary thromboembolism. A case report and review of the literature]. AB - A 39-years-old male patient with chronic venous insufficiency, deep venous thrombosis and recurrent pulmonary embolism in the past medical history. After syncopal event was diagnosed of bilateral chronic pulmonary embolism, pulmonary hypertension and right ventricular failure. Fibrinolytic treatment was no effective therapeutic modality. Under cardiopulmonary bypass, bilateral pulmonary thromboendarterectomy with extension into lobe arteries, plus insertion of caval filter was performed. We present our experience with this case and a review of the literature. PMID- 7481041 TI - [Evolving identities of bronchopulmonary adenocarcinomas]. PMID- 7481042 TI - [Chemosensitivity and sleep apnea syndromes]. PMID- 7481043 TI - [Lung cancer in young subjects: implications for epidemiological and etiological research]. PMID- 7481044 TI - [Palliative treatment of bronchial obstruction by tumor. For whom? Why? How?]. PMID- 7481045 TI - [Cytokine inhibitors in intensive care]. AB - OBJECTIVE: To examine three typical disease states seen in intensive care, sepsis, Fulminant purpura and acute respiratory distress syndrome (ARDS) to assess the implication of cytokines in their pathogenesis and particularly in the clinical applications of possible cytokine inhibitors. SOURCE OF DATA: The data bank of MedLine and the Index Medicus 1985-1993 and the first part of 1994. These sources have enabled us to consult publications in French and English and to include information on both animals and humans. The publications issued from Intensive Care Congresses have also been scrutinised; the Societe de Reanimation de Langue Francaise, The American Thoracic Society and the 13th and 14th International Symposium on Intensive Care and Emergency Medicine, Brussels. SELECTION OF DATA: This review emphasizes certain areas of work including recognised work which has been published on the immunotherapy of sepsis in man, on those papers which have been published as a preliminary communication in the from of a summary and on certain papers relating to animal work which are regularly cited in Intensive Care literature. DISCUSSION: The relationship between cytokines and the three selected disease states have been briefly described. The greater part of those papers which have either been published or are in the process of being published present pharmacotherapeutic data in phase 2 or phase 3 in relationship to anticytokines and sepsis. As for the treatment of Fulminant purpura and ARDS, using anticytokine antibodies in 1994 we are still in the stage of hypothesis and speculation. CONCLUSIONS: Future clinical strategies designed to combat. Future clinical strategies designed to fight against the most critical diseases in intensive care medicine require some use of any kind of immunotherapy. In animal studies, convincing data are available showing that immunotherapy improves the prognosis of sepsis, whereas in humans, to date, the results appear to be deceiving. Future research in this direction is mandatory, in sepsis and in other disease states, like ARDS, because no other hope for treating these patients seems to appear in a near future. PMID- 7481046 TI - [Metrology of airborne particles carrying the principal air allergens]. AB - The relationship between exposure to aeroallergens and the acquisition of allergy and asthma has been shown over the past ten years. Thanks to new developments for detecting major allergens amongst the principle aeroallergens, in future it will be possible to measure their airborne concentration and to determine the particle size of particles carrying them. We report the results obtained from three studies in which we have shown that airborne mites Group I and II and cockroaches Bla g 1 and Bla g 2 allergens have a broadly similar airborne behaviour. That is to say that they are not measurable unless the atmosphere is artificially disturbed and they are carried principally on particles of > 10 microns. On the other hand, 30-40% of cat allergens are carried on particles < 5 microns and measurable in the air without any artificial disturbance. According to our data and those of the literature, we propose a different classification for aeroallergens according to characteristic aerodynamics. Although some progress in the standardisation of techniques for sampling airborne allergens may have been accomplished, other studies are required to improve their reliability in order that airborne measurements can become a marker of allergic exposure in both domestic and occupational environment. PMID- 7481048 TI - [Study of chemosensitivity in patients believed to have sleep apnea syndrome]. AB - We performed polysomnography and measured hypoxic ventilatory (HVR), hypercapnic ventilatory responses (HCVR) in 42 patients (60 +/- 11 years) with obesity and a clinical suspicion of sleep apnea syndrome (SAS) in order to determine whether an altered chemosensitivity was associated with SAS. The apnea/hypopnea index was 38 +/- 20 events per hour of sleep in 28 patients (SAS+ group) and less than 10 in the 14 others (SAS- group). The 2 groups differed only by a lower waking PaO2 in SAS+ as compared to SAS- (71.0 +/- 9 vs 77.4 +/- 8 mmHg, p < 0.05). HVR and HCVR were not significantly different in the 2 groups (0.82 +/- 0.58 vs 0.86 +/- 0.37 l.min-1.%-1; 1.41 +/- 0.81 vs 1.40 +/- 0.67 l.min-1.mmHg-1, respectively). In SAS+ group, HVR or HCVR did not change 3 or 12 months after continuous positive airway pressure (CPAP) therapy while both polysomnography and PaO2 returned to normal. We conclude that in patients with mild obesity and SAS there is no difference in chemosensitivity due to the presence of sleep apnea and that CPAP therapy does not alter these measurements. These results suggest no direct effect of SAS on chemosensitivity in the population studied. PMID- 7481049 TI - [Evolvements in the presentation of primary bronchial adenocarcinomas observed in a pneumology service over an 11-year period]. AB - The evolution of the clinical presentation, endoscopy and radiology in a consecutive series of 121 cases of primary bronchial adenocarcinoma has been studied in a retrospective manner over a period of 11 years. The mean age was 60 +/- 12 and 16.5% of the patients were women. The frequency of smoking and the quantity smoked in packed years was significantly higher in men than in women. The frequency of the presenting signs and the endoscopic features did not change over the period. On the other hand, the peripheral and mediastinal-hilar forms were more frequent from 1990 in a significant manner so that the hilar form became rarer. The result seems to confirm an increase in the frequency of a new radio-clinical expression of primary bronchial adenocarcinoma. These mediastinal hilar adenocarcinomas do not have specific characteristics but are associated with a higher mortality. PMID- 7481047 TI - [Epidemiology of malignant pleural mesothelioma in the Nantes-Saint Nazaire region. Course in 1956-1992]. AB - The aim of this study was to estimate the evolution of the incidence of mesothelioma in the Loire-Atlantique department since 1956 and to register all the cases diagnosed between 1985 and 1992 and to complete the data of earlier studies carried out between 1956 and 1984. The cases were indexed from the files of the pathology department and also from demographic and occupational data concerning the patients which had been gathered retrospectively from an inquiry of patients attached either to private physicians or to a hospital service. From 1956 to 1984 there were 125 cases (119 men, 6 women) who had been diagnosed; 92 cases were registered between 1985 and 1992 (79 men and 13 women). An increase in the annual number of cases was significant. The incidence during the period 1985 1992 was 10.9 per million inhabitants (men 19.4, women 3) against 8.7 (men 17.2, women 0.8) for the period 1975 to 1984 and 2.6 (men 5.2, women 0.2) for the period 1956-1974. The mean age of the subjects at the time of diagnosis rose during the period studied (59.2 +/- 9.4 between 1956-1974 to 63.1 +/- 11.9 between 1975-1984 and 67.0 +/- 9.7 between 1985-1992). Occupational exposure to asbestos was certain or probable in 85 per cent of cases with a median duration of exposure of 25 years (range 2 months to 48 years) with a median interval between the first exposure to diagnosis of 44 years (range 10-70 years). The industrial sector most often implicated was naval construction (127 cases).(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7481050 TI - [Diagnostic and therapeutic strategies in exacerbations of chronic bronchitis in city practice]. AB - The consensus conference convened by the French Language Society for Infectious Disease at Lille in 1991 stressed the fact that two germs were most often the cause of exacerbation in chronic bronchitis (Streptococcus pneumoniae and Haemophilus influenzae) and that antibiotic therapy was the "safe solution" and that the first intention treatment should be either penicillin A, a first generation cephalosporin or a macrolide for the first 8-10 days. A chest x-ray was recommended if there was the slightest doubt about co-existing parenchymal disease with a reevaluation around the 7th day and a prescription of penicillin A plus a beta-lactamase inhibitor or a second or third generation cephalosporin in case of failure. The aim of this study was to assess the diagnostic and therapeutic attitudes of general practitioners when faced with exacerbation in chronic bronchitis in an adult of 60 without severe signs and to find out the antibiotic of first choice and also the antibiotic to be used if the first treatment failed. One hundred doctors were drawn at random from a list of general practitioners in Bordeaux. They were requested to reply to a questionnaire on the strategy of first choice antibiotic and the means of reassessment of the treatment after it had been instituted and the strategy used when faced with a patient who did not improve after the initial treatment.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7481053 TI - [Pleural fibroma. Contribution of immunohistochemistry]. AB - The authors report a case of pleural fibroma discovered during an examination for thoracic pains in a 65 year old woman. Histochemical studies were positive with vimentin and negative with KL1 antibodies and compatible with the most recent hypotheses concerning the purely fibroblastic origin of these tumours. PMID- 7481051 TI - [Presumed transudative pleural effusion?]. AB - We report a case of recurrent transudative pleural effusion. The initial cardiovascular investigations failed to determine its cause. The catheterization showed a "dip-plateau" suggesting a restrictive cardiomyopathy. Endomyocardial biopsy finally proved a cardiac amyloidosis. PMID- 7481052 TI - [Disseminated aspergillosis with pulmonary and cardiac localization in an HIV infected patient]. AB - The authors report a case of invasive aspergillosis with pulmonary and cardiac involvement revealed by a pericarditis in a seropositive patient (VIH1). Clinical findings, differential diagnosis and outcome underly the issues encountered in the management of aspergillosis in AIDS patients. Indeed, the low prevalence of aspergillosis in AIDS and the insidious features of aspergillosis locations make the early diagnosis difficult. It is though essential for a favourable outcome under specific treatment. The risk factors, clinical characteristics and diagnostic procedures are mentioned as well as the various available treatments. PMID- 7481055 TI - [Dr. Freitag's dynamic tracheobronchial prosthesis (or dynamic stent)]. PMID- 7481054 TI - [Diagnostic difficulties in angiosarcoma of the pulmonary artery with hemoptysis]. AB - Intimal sarcomas growing from the pulmonary trunk or branches of the pulmonary artery, are rare tumours in which the diagnosis is most often made at autopsy or during a thoracotomy. Usually the clinical pictures is non specific resembling a severe pulmonary embolus which is resistant of all treatment. With the help of new imaging techniques, a pre-operative diagnosis is made in more than half the cases. When there is a tumour which is relatively localised and without endoluminal invasion, as in the observation reported here, the diagnosis rests on the histology from the operative specimen. PMID- 7481056 TI - [Cylindric intrabronchial support made of silicone Endoxane (Pr Dumon)]. PMID- 7481057 TI - [A double radiological curiosity]. PMID- 7481058 TI - [Respiratory syncytial virus infection in adults]. PMID- 7481059 TI - [References for evaluation instruments in quality assurance in rehabilitation--3. The EAM profile system--capacity and requirement profile as a decision aid in the tension field between (medical) rehabilitation and vocational integration]. AB - The EAM profile system is characterized by enabling comparison of a subject's abilities with the requirements present on the job. The article initially outlines the structure of the necessary abilities and/or requirements profiles, describes the assessment criteria used, and concludes by presenting a case example, showing that the question of individual employability and placement can be answered by acting upon profile comparison findings. PMID- 7481060 TI - [Different phases in neurologic rehabilitation. Association of German Pension Insurance Carriers]. AB - Elaborated by the project subgroup for "Neurological Rehabilitation" of the Federation of German Pension Insurance Institutes, the phase model presented describes the phases of neurological care and rehabilitation for patients with neurological conditions (notably stroke, craniocerebral trauma, and multiple sclerosis). It deals with acute and intensive care, inpatient medical rehabilitation, as well as the phase of aftercare and the care/rehabilitation phase where long-term support, attendant and/or maintenance measures are required (outpatient or partial-hospitalization medical rehabilitation modalities are not included as yet). Based on patient characteristics, care and rehabilitation goals are specified along with the care and rehabilitation tasks that can be derived from these, with great diversity of individual management course being possible. PMID- 7481061 TI - [Neuropsychological analysis of schizophrenia-induced attention deficits--attempt at an overview]. AB - In this review schizophrenic attention disorders are described from a neuropsychological perspective. After a historical excursion to Wundt, James and Kraepelin, the experimental psychology attention research of the last decades is presented, with special regard to Joseph Zubin's model of attention. After describing the symptomatology and course of schizophrenic psychosis, the concept of positive and negative symptoms according to Crow and Andreasen is set out as well. In describing the cognitive disorders it is on the one hand tried to elucidate the concept of "Basisstorung", and, on the other, to deal with attention disorders as potential vulnerability markers. After setting out various models re. the origin of attention disorders in both acute and chronic psychoses, implications for diagnosis and treatment are offered for the clinical psychologist in conclusion. PMID- 7481064 TI - [Attitude of physicians to rehabilitation--an empirical study of rheumatologists]. AB - Although the medical doctor holds a central steering function in making rehabilitation referrals, hardly any empirically founded information is available on physicians' attitudes towards rehabilitation, their indication considerations and actual referral practice. Prompted essentially by this information deficit, an anonymous questionnaire study was undertaken with 49 rheumatologists concerning their knowledge and experience in rehabilitation, their views of various forms of rehabilitation (inpatient, partial-hospitalization, out-patient) as well as medical further education in rehabilitation subjects. Reflecting a positive basic view of rehabilitation, nearly half of the responding rheumatologists subjectively felt well-informed about rehabilitative issues, with 49% stating that they frequently recommended patients for rehabilitation. As far as preferences are concerned in terms of community vs. non-community rehabilitation concepts, priority is given by 39% to partial-hospitalization rehabilitation ("most favourable"); more than 75% of the interviewees however advocated case-by-case decision-making. Accordingly, only 18% consider ambulatory or day centre programmes true alternatives for inpatient rehabilitation, and 78% rather consider them complementary to each other. The advantage of partial hospitalization rehabilitation close to the rehabilitee's place of living is seen above all in potential involvement of relatives and family doctor. Aspects stated in favour of non-community inpatient rehabilitation primarily are getting away from stressful family surroundings and the comprehensive medical programme available. A strong need for rehabilitation-related further education is expressed, with forms of rehabilitation, psychosocial rehabilitation, and access modes among the primary contents to be addressed. PMID- 7481063 TI - [Dermatologic climate therapy--definition, indications and public health necessity]. AB - Dermatological climatotherapy is used in the treatment of chronic and chronically relapsing long-term dermatoses, such as, especially, atopic dermatitis and psoriasis, complementing the dermatological therapies applied at the patient's place of living. It is a classical dermatological in-clinic therapy carried through in specific rough and stimulating climate areas of proven therapeutic benefit, primarily the North Sea climate and certain alpine locations over 1 500 m above sea-level, like especially in the high mountain valley of Davos, Switzerland. Dermatological climatotherapy is a well-tried therapeutic agent which, in comparison to the dermatological therapies applied at home, as a rule has fewer side effects as well as, above all, an additional long-lasting time effect. The latter is of particular importance regarding chronical diseases and cannot be achieved by other therapies. Dermatological climatotherapy, hence, represents an all-entailing therapy form and moreover is the most comprehensive therapy anyway. Dermatological climatotherapy can only be performed in rough and stimulating climate zones with additional special insolation, thermichygric and defined aerosol conditions. On the one hand, the organism is stabilized general by those climatic conditions, and, on the other, the constitutionally damaged skin is affected positively by the direct influence of climate factors. As every climate has its specific overall effect, the therapeutical immediate and long term effects of a climate have to be proven scientifically by follow-up studies. PMID- 7481062 TI - [Neuropedagogic aspects of children with acquired craniocerebral trauma--an overview]. AB - In this article a review is given of the neuropedagogical aspects of children with acquired head injury (cerebral concussion). For this purpose attention has been paid to a study of the literature in the period 1970-1991. The attention has been focused on the most frequently occurring disorders such as cognitive and other neuropsychological disorders, behavior disorders and the post-traumatic syndrome. It appeared that the views about the origin of the disorders are very diverse. This applies in particular to cognitive and other neuropsychological disorders and behavior disorders. A consensus for the existence of a post traumatic syndrome has not been found. PMID- 7481066 TI - ["Profession-oriented project work" in preparation for occupational rehabilitation]. AB - In the framework of pre-rehabilitation courses for German-origin immigrants as well as other rehabilitees of mostly foreign nationality, the Bad Krozingen cardiac care centre has for some two years, i.e. since 1993, undertaken practical trials of a model occupational field-oriented project classroom. To enable rehabilitees to better prepare for their future work or occupational fields, internal differentiation is used to facilitate occupation-specific preparation in a project worksetting, with self-directed activity of the individual in a teamwork situation being the main focus of enhancement. This afternoon "occupational field-oriented project work" complements morning classrooms that, aside various other subjects, are focussed on German and maths as well as EDP fundamentals, with electronic text processing in particular viewed as a tool that facilitates written documentation of teamwork, hence is dealt with right at the beginning of the course. PMID- 7481065 TI - [Naturopathy in geriatric rehabilitation]. AB - In the treatment of acute disorders, the complex interactions existing among various geriatric illnesses and age-specific impaired organ functioning, demand a therapeutic outlook oriented towards essential aspects, which can, and should, however be expanded in a holistic sense during rehabilitation. In this context, classic naturopathic techniques offer methods of treatment highly suitable in particular for geriatric patients with reduced general condition, methods that are readily accepted by them and available from the areas of kinesitherapy, hydrotherapy, phytotherapy, dietetics, and the restoration of harmonic life patterns and rhythms. PMID- 7481067 TI - Properties and regulation of pH-dependent cation channels in the apical membrane of cultured proximal tubule cells. AB - The established opossum kidney (OK) cell line serves as a model system for ion and substrate transport in the renal proximal tubule. Previous experiments on OK cells revealed a channel-mediated Na+ conductance which is regulated by intracellular pH (pHi). In this study we report on patch clamp experiments determining the properties and pHi dependence of a cation channel located in the apical membrane. This channel is selective for sodium over chloride but discriminates poorly between the monovalent cations Na+,K+,Li+ and Cs+. Its open probability (P(o)) rises at depolarising membrane potentials. Under normal conditions the channel is inactive in the cell-attached patch mode and is activated upon excision. However, after excision the channel usually runs down within 30-90 s which cannot be overcome by either altering the Ca(2+) concentration (10(-3) mol/l, 10(-6) mol/l, Ca(2+)-free) or adding 1 mmol/l Mg-ATP to the bath solution. In the cell-attached patch mode the channel could be activated by decreasing pHi from pH 7.4 to pH 6.5, by either the ammonium prepulse technique or the nigericin K+ method, in more than 50% of the experiments performed. In the renal proximal tubule such a non-selective cation channel would constitute a functional Na+ channel and might therefore support Na+ reabsorption especially during the intracellular acidification due to hormonal inhibition of the Na+/H+ exchanger. PMID- 7481068 TI - Effects of glyburide on renal tubule transport and potassium-channel activity. AB - Renal clearance, microperfusion and patch-clamp techniques were used to investigate the effects of the K-channel blocker glyburide on electrolyte excretion, the transport properties of the thick ascending limb (TAL) of Henle and K-channel activity in the apical membrane of the TAL and of the cortical collecting tubule. Our data suggest that the K-channel blocker glyburide can inhibit transport of Na and K in the TAL by blocking K recycling across the apical membrane. Additionally, inhibition of K secretion in the collecting ducts occurs by decreasing the activity of apical K channels and prevents kaliuresis. PMID- 7481069 TI - Effects of enalapril treatment on gene expression of smooth muscle myosin heavy chain isoforms in glomeruli of diabetic rats. AB - We have previously shown that smooth muscle contains three types of myosin heavy chains: SM1, SM2, and SMemb. The present study was designed to assess how glomerular expression of mRNA for these isoforms is regulated and whether their expression is affected by enalapril treatment in diabetic rats. Animals were divided into 4 groups: (1) untreated diabetic rats; (2) enalapril-treated diabetic rats; (3) untreated control rats, and (4) enalapril-treated control rats. Enalapril treatment was continued for 24 weeks. The glomerular mRNA levels for SM1 and SM2 showed little change in all groups throughout the experimental period. In contrast, SMemb mRNA in group 1 increased significantly with age compared to levels found in untreated controls (4.6-fold higher at 4 weeks, p < 0.01; 6.8-fold higher at 12 weeks, p < 0.01, and 10.6-fold higher at 24 weeks, p < 0.001). Enalapril reduced both creatinine clearance (p < 0.01) and urinary protein excretion (p < 0.01) in diabetic rats. Moreover, enalapril significantly attenuated the increase in the glomerular SMemb mRNA level in diabetic rats (the difference between treated and untreated rats was significant at p < 0.01 from week 4 to 24). However, enalapril had no effect on SMemb mRNA levels in controls. These data suggest that SMemb is a molecular marker for phenotypic alteration and that the beneficial effect of enalapril on proteinuria and renal function may be, at least in part, associated with reducing SMemb mRNA expression in diabetic glomeruli. PMID- 7481070 TI - Mercapturic acid formation in cultured opossum kidney cells. AB - We investigated the last step of mercapturic acid formation, the N-acetylation of cysteine S-conjugates, in the established opossum kidney (OK) cell line which exhibits characteristics of the proximal tubule. S-Benzyl-L-cysteine was used as a model substance for such a cysteine S-conjugate. We succeeded in showing that OK cells absorb S-benzyl-L-cysteine via an active transport system which is inhibitable by phenylalanine. This transport follows Michaelis-Menten kinetics and the two characterizing parameters were determined: the Michaelis-Menten constant Km = 1.8 mmol/l, and the maximum of the difference between the intracellular and the extracellular concentration of S-benzyl-L-cysteine delta Cmax = 19.4 mmol/l. S-Benzyl-L-cysteine is converted to N-acetyl-S-benzyl-L cysteine at a constant rate, which is independent of the extracellular S-benzyl-L cysteine concentration. Under the tested experimental conditions this is probably due to saturation of the microsomal N-acetyltransferase catalyzing this reaction. In conclusion, we have shown that OK cells are a suitable model for studying mercapturate formation. They take up S-benzyl-L-cysteine mainly via the same carrier as phenylalanine, which is known to be transported in the rat by the high capacity, low-affinity neutral amino acid carrier, and convert it to N-acetyl-L benzyl-S-cysteine. PMID- 7481071 TI - Nitric oxide does not contribute to sodium retention and peripheral vasodilation induced by partial portal vein ligation in rats. AB - The role of nitric oxide (NO) synthesis in the peripheral vasodilation and sodium retention that occurs after partial portal vein ligation (PVL) was investigated. Hemodynamic studies in PVL rats with sodium retention and in sham-operated controls were conducted on the day when PVL rats developed transient and maximal sodium retention. Measurements were obtained before and during two consecutive periods after NO synthesis inhibition with NG-monomethyl-L-arginine (L-NMMA). Under baseline conditions, PVL rats with sodium retention were hypotensive, with equivalent decreases in total peripheral resistance and glomerular filtration rate in comparison to the control group. After L-NMMA, peripheral resistance and arterial pressure increased by similar extent in both groups. As compared with controls, PVL rats with sodium retention remained hypotensive and vasodilated. Furthermore, L-NMMA-induced natriuresis was attenuated in the PVL group. Additionally, serum and urinary levels of nitrate and nitrite did not vary before surgery and at the time of sodium retention. These results suggest that in PVL rats (1) vasodilation is not NO mediated; (2) vasodilation is not a sufficient explanation for sodium retention, and (3) a sodium-retaining factor acting on the renal tubules is responsible for sodium retention. PMID- 7481074 TI - Inhibition of in vitro HIV infection by trinitrophenyl-protein conjugates. AB - Levels of natural antibodies (NAb) with high anti-trinitrophenyl (TNP) activity are increased during human immunodeficiency virus (HIV) infection. The aim of the present study was to examine the anti-HIV effect of natural anti-TNP antibodies, as well as that of their internal image, TNP antigen, on HIV infection in vitro. The results obtained with anti-TNP antibodies, as assessed by syncytia formation, were variable, although they demonstrated an inhibitory effect. In contrast, using RT activity assay plus evaluation of syncytia formation and the viral cytopathic effect, we found that bovine serum albumin (BSA) bearing different TNP groups was able to inhibit HIV infection of peripheral mononuclear cells and T4 cell lines without affecting cell metabolism or proliferation. BSA alone was devoid of activity; the antiviral effect depended on TNP substitution of the BSA molecule, and passage through an anti-TNP immunoadsorbent abolished this effect. The mechanism by which TNP exerts this antiviral effect is unclear. Antigenic epitopes may be shared by HIV and TNP, since monoclonal antibodies directed against various HIV proteins reacted with TNP in an enzyme immunoassay. TNP-BSA, however, did not bind to the CD4 receptor. PMID- 7481073 TI - Cyclosporine-induced apoptosis in CD4+ T lymphocytes and computer-simulated analysis: modeling a treatment scenario for HIV infection. AB - Cyclosporine A (CsA) is a potent immunosuppressive drug which interferes in vitro and in vivo with T-cell function. CsA has been shown to arrest T-cell maturation intrathymically and to inhibit T-cell proliferation. In this study, we demonstrate that CsA induces apoptosis in the canine CD4+ CD8- T-lymphocyte cell line 401 in a dose- and time-dependent fashion. Similar results could also be obtained from human peripheral blood lymphocytes. Apoptosis is observed within 4 hours after CsA application and is not prevented by excessive addition of ConA supernatant as a source of interleukins. The induction of apoptosis in CD4+ T lymphocytes suggests a possible treatment option for human immunodeficiency virus (HIV) infections, since the major target population for the HIV would be ablated at short term. A computer-simulated analysis with the "Cybermouse" HIV model confirmed that the virus would eventually disappear and HIV-infected macrophages would also be substantially reduced if CsA were given in combination with drugs which block viral replication (3'-azido 3'-deoxythymidine or 2',3' dideoxycytidine). This treatment scenario could be applied under controlled conditions and with supportive patient care. A further review of the literature also suggests the positive impact of CsA treatment on the progression and outcome of AIDS-related mortality. PMID- 7481072 TI - Micropuncture study of the effect of furosemide on proximal and distal tubules of the rat nephron. AB - The tubular effects of furosemide were studied by micropuncture and clearance techniques on 20 rats. Collections of tubular fluid (TF) from early distal (ED) and late proximal (LP) segments of the same nephrons and of different nephrons were performed during baseline conditions. Re-collections were taken from the same sites and new collections from different nephrons after 10 mg/kg furosemide. The glomerular filtration rate (GFR) was 1,309 +/- 212 microliters/min during baseline, and 1,348 +/- 199 microliters/min after furosemide (p > 0.89); while the urine flow rate rose from 36 +/- 8 to 167 +/- 30 microliters/min (p < 0.001). The nephron filtration rate (NFR) was not different in 46 paired distal (33.3 +/- 2.6 nl/min) versus proximal samples (34.2 +/- 2.9 nl/min, p > 0.72), neither was it different during baseline (37.2 +/- 1.4, n = 120) as compared to furosemide (37.2 +/- 2.7, n = 91, p > 0.99). The percent reabsorption (PR) at the ED sampling site was 87 +/- 4% during baseline, and 89 +/- 3% in 13 paired samples during furosemide (p > 0.47). PR at the LP sampling sites was 83 +/- 2% during baseline, and 80 +/- 2% in 26 paired samples during furosemide (p > 0.63). In 31 paired ED-LP collections, PR was 82 +/- 4 (ED) versus 72 +/- 4% (LP) during baseline, and 87 +/- 3 versus 74 +/- 6%, respectively, during furosemide. The respective collection rates were 4.6 +/- 1.0 versus 9.5 +/- 1.3 nl/min during baseline (p < 0.0001), 5.8 +/- 2.3 versus 8.7 +/- 3.0 nl/min during furosemide. The LP-ED differences obtained during baseline were not different from those measured during furosemide for the collection rate, PR and NFRs. The absolute LP resorption rate was not significantly different during baseline as compared to furosemide. Thus, furosemide did not affect the difference between ED and LP collection sites in collection rate, absolute and fractional reabsorption, in the absence of changes in GFR and NFR. These data indicate that furosemide acts solely along Henle's loop, where it blocks Na+ transport. The urine flow rate rises during furosemide because water abstraction along the distal tubule is reduced by the isotonicity of ED TF, and along the collecting ducts by the isotonicity of the medullary and papillary interstitium caused by the diuretic. We conclude that under the conditions of the present study, furosemide has no proximal effect. PMID- 7481076 TI - Expression of neutrophil activation markers and neutrophil adhesion to chondrocytes in rheumatoid arthritis patients: relationship with disease activity. AB - Neutrophil infiltration in synovial fluid is an important step in inflammation characterizing rheumatoid arthritis (RA). In this study, the activation and functional state of neutrophils in the blood and synovial fluid of patients with rheumatoid arthritis were compared: mean density of neutrophil activation markers CD11b, CD18 and L-selectin was measured with a flow cytometer, and adhesion to chondrocytes using a fluorimetric assay. No significant differences between control and patient peripheral blood neutrophils were observed. When comparing neutrophils of patient peripheral blood with paired synovial fluid, an increase in CD11b (p = 0.008) and a decrease in L-selectin (p = 0.008) were measured. For neutrophils of control and patient peripheral blood, fMet-Leu-Phe stimulation induced upregulation of CD11b (resp p = 0.007 and p = 0.008) and CD18 (resp p = 0.005 and p = 0.01). In the synovial fluid, no significant increase in CD11b and CD18 could be induced with fMet-Leu-Phe. Percentages of adherent neutrophils were comparable between controls and patients, both in peripheral blood and synovial fluid. Adhesion to chondrocytes of peripheral blood neutrophils of patients was correlated with clinical (Ritchie) and biological (erythrocyte sedimentation rate) parameters (resp r = 0.67, r = 0.73). In conclusion, these results demonstrate that peripheral blood neutrophil adhesion to chondrocytes was correlated with active disease, and that synovial fluid neutrophils were activated in vivo. These findings provide further evidence for the contributing role of neutrophils in articular destruction in RA. PMID- 7481075 TI - T-cell repertoires in healthy and diseased human tissues analysed by T-cell receptor beta-chain CDR3 size determination: evidence for oligoclonal expansions in tumours and inflammatory diseases. AB - Many examples of oligoclonal T-cell expansion in infiltrated diseased tissues have been reported. However, it remains to be established whether such observations can be generalized and to what extent oligoclonal patterns obtained after in vitro culture of T-cell infiltrates reflect in vivo situations. Using new high resolution analysis which requires no in vitro cellular expansion, we detected such oligoclonal T-cell expansions in 7/7 melanoma tumour biopsies, 3/3 biopsies of inflammatory skin during acute graft versus host disease (aGVHD) after allogeneic bone marrow transplantation (alloBMT) and 7/7 synovial membranes from patients with rheumatoid arthritis. Thus, oligoclonal T-cell expansions are readily observed when a sufficiently sensitive detection method is used, suggesting that similar expansions are the rule among T-cell infiltrates in different diseases. This observation and the monitoring of the in vivo evolution of such expansion during the course of the disease and during in vitro culture should have important clinical implications. PMID- 7481078 TI - On becoming skillful: patterns and constraints. PMID- 7481077 TI - Partial inhibition of Theileria parva-infected T-cell proliferation by antisense IL2R alpha-chain RNA expression. AB - Antisense RNA expression was used to assess the role of the interleukin 2 receptor (IL2R) alpha-chain in proliferation of T cells transformed by infection with the intracellular parasite Theileria parva. Two vectors were constructed, in which part of the bovine IL2R alpha-chain cDNA was inserted in either a sense or antisense orientation in the plasmid pHS1-CAT, which is expressed by induction of the human metallothionein IIA (hMT-IIA) with cadmium (Cd2+). A T. parva-infected T-cell line, TpM(803), known to express the IL2 and IL2R genes in a constitutive way, was permanently transfected with one of the two constructs, and the effect of antisense IL2R alpha-chain RNA expression upon proliferation of TpM(803) cells was tested. Antisense-transfected TpM(803) cells grew much more slowly than sense transfected cells even in the absence of added Cd2+, but the addition of Cd2+ to the culture medium resulted in further inhibition. A strong reduction in IL2R alpha-chain mRNA was observed in antisense but not in sense-transfected cells. These observations confirm a role for the IL2R alpha-chain in proliferation of T. parva-infected TpM(803) cells. PMID- 7481079 TI - Children's activity levels and lesson context during third-grade physical education. AB - Little is known about children's activity levels during physical education classes and how they relate to the national health objectives for the year 2000 (U.S. Public Health Service, 1991). We systematically observed students' physical activity and associated variables in 293 third-grade physical education lessons in 95 schools in 4 Child and Adolescent Trial for Cardiovascular Health (CATCH) centers in California, Louisiana, Minnesota, and Texas. The influence of independent variables was assessed simultaneously by analysis of variance. Significant differences among study centers were evidenced for both physical activity and lesson context variables. Additionally, there were differences among variables for lesson location and teacher specialty, but not for teacher gender. Boys were more active than girls, but only during free play opportunities. The findings have widespread implications for educators responsible for developing and implementing health-related physical education programs, as well as for designing and conducting staff development. PMID- 7481080 TI - Expert-novice differences in cognitive and skill execution components of youth baseball performance. AB - Previous research has shown that experts exhibit superior response selection and skill execution during performance in youth sport. The purpose of this study was to examine differences in cognitive and skill execution components of game performance in young baseball players (N = 159) with varying levels of expertise. Three levels of expertise (low-, average-, and high-skilled players) were identified at each age level (7, 8, 9, and 10 years of age). Game performance was videotaped, and measures of skill execution (throwing accuracy, throwing force, fielding, catching, batting average, and batting contact) and cognitive components (positioning, decisions) were developed from observational analysis. The results indicated that baseball skill execution during game play maximally discriminated expertise levels. PMID- 7481081 TI - The validity of the Tritrac-R3D Activity Monitor for the assessment of physical activity in children. AB - The purpose of this study was to evaluate the validity of the Tritrac-R3D Activity Monitor, a new instrument designed to improve assessments of physical activity. Comparisons were made with a heart rate monitor and with a Caltrac Activity Monitor. Thirty-five children (ages 9-11 years) were monitored on 3 different school days with all 3 instruments. The Tritrac was moderately correlated with the heart rate monitor (r = .58) and highly correlated with the Caltrac monitor (r = .88). By taking advantage of the minute-by-minute timing capability of the Tritrac and the heart rate monitors, it was discovered that the correlations between these instruments were highest during free play situations (lunch/recess, recess, after school) and were lower when activity was more limited (class time) or structured (physical education). The ability of the Tritrac to assess activity on a minute-by-minute basis may greatly enhance its overall utility. PMID- 7481082 TI - The manipulation of vision during the powerlift squat: exploring the boundaries of the specificity of learning hypothesis. AB - The available information for controlling a multidegree-of-freedom sport action was manipulated in 2 experiments. In the first, 10 intermediate lifters were participants; for the second, 8 skilled and 8 less skilled lifters were observed. Three single repetitions of a powerlift squat were performed under 3 vision conditions (i.e., full, ambient, no vision). The less skilled and intermediate lifters' technical performance decreased significantly with the removal of visual information. There was no detrimental effect in the skilled group. Despite the differing information constraints, skilled lifters exhibited a high level of positioning accuracy and timing consistency across conditions. These data fail to support the theoretical predictions of the specificity of learning hypothesis. The differences between the task constraints in this study and those in manual aiming investigations may represent a boundary to the current propositions of the specificity of learning hypothesis. PMID- 7481083 TI - Value orientations of preservice teachers. AB - Curriculum theorists have acknowledged the critical role that beliefs and values play in the decisions that teachers make, but very little is known about how teachers' value profiles develop. The purpose of this investigation was to examine the educational value orientations of a group of physical education preservice teachers (N = 16) enrolled in an elementary methods course and to investigate the link between value profiles and teacher behaviors. Value profiles were assessed using the Value Orientation Inventory (Ennis & Hooper, 1988). The results indicated that teachers in training entered field-based experiences with defined value profiles, but that these profiles were not stable constructs. Priorities within competing value orientations fluctuated over the course of a semester. Possible explanations for the changes that occurred and implications for teacher education programs are discussed. PMID- 7481084 TI - The effects of cadence, impact, and step on physiological responses to aerobic dance exercise. AB - The physiological responses to aerobic dance exercise of varied impact (high, low), step (less arm movement vs. more arm movement), and cadence (124 vs. 138 beats.min-1) were investigated. Experienced, female aerobic dancers (N = 16) performed activities that combined the levels of impact and step for 3 trials of 8-min each. Dependent variables included heart rate, percentage of maximal heart rate, oxygen consumption, percentage of maximal oxygen consumption, and respiratory exchange ratio. Repeated measures analyses of variance indicated a significant Impact x Step interaction whereby oxygen consumption was greater for the high impact-less arm movement activity (jog), while the low impact-more arm movement activity (power jack) was greater for heart rate. The interaction of aerobic dance characteristics (e.g., impact, arm movement) that may alter physiological responses to aerobic dance exercise should be identified in future aerobic dance routines and studies. PMID- 7481085 TI - Physiological and perceived exertion responses to six modes of submaximal exercise. AB - In order to compare the cardiovascular and perceived exertion responses to 6 modes of exercise, 10 male recreational exercisers were habituated to treadmill jogging, stationary skiing, shuffle skiing, stepping, stationary cycling, and stationary rowing. After following a specific dietary preparation, each participant performed a 20-min exercise bout at a constant 14-rating of perceived exertion (RPE) followed by a second exercise bout at 60% of mode-specific peak oxygen consumption (VO2peak). On the 14-RPE trial, oxygen consumption (VO2) and oxygen pulse were significantly higher during jogging than during other exercise modes, and oxygen pulse was higher during skiing than during shuffle skiing. On the 60% VO2peak bout, oxygen pulse was significantly higher during jogging than during shuffle skiing, cycling and rowing. Ratings of perceived exertion were significantly higher during cycling than during jogging. These results indicated that a variety of exercise modes can be used to develop fitness, but jogging may induce a slightly more favorable VO2-to-RPE relationship. PMID- 7481086 TI - Fair Play for Kids: effects on the moral development of children in physical education. AB - Commitment to the principles of sportspersonship is an acknowledged goal for school physical education. However, few programs have been implemented to investigate moral development changes in physical activity settings. A field experiment was designed to examine the effect of participation in educational activities selected from Fair Play for Kids (1990) on the moral judgment, reason, intention, and prosocial behavior of children (N = 452) in the 4th through 6th grades. Six intact classrooms at each grade level (N = 18) were randomly assigned to the following groups: (a) control, (b) Fair Play for Kids curriculum during physical education only, or (c) Fair Play for Kids curriculum during all school subjects. Experimental protocol extended for 7 months of an academic year, and moral development indicators were assessed prior to and following the intervention. Using class as the unit of analysis, 3 x 2 (Group x Time) repeated measures analyses of variance revealed that both treatment groups were significantly higher than the control group at posttest for moral judgment, reason, and intention scores. For students within classes, repeated measures analyses showed that treatment group participants had significantly higher posttest scores on all 4 measures as compared to students in the control group. Results provide initial validation of the Fair Play For Kids curriculum for effecting change in the moral development of elementary school students. PMID- 7481087 TI - Relationships of age and muscle mass to peak torque in high school wrestlers. PMID- 7481088 TI - Does mental practice work like physical practice without information feedback? PMID- 7481089 TI - Residual expression of reporter genes in constructs mimicking HIV genome organization. AB - Plasmids were constructed whereby the expression of a reporter gene, either the cDNA corresponding to the secreted form of human alkaline phosphatase (SEAP) or the herpes simplex virus type 1 (HSV1) thymidine kinase (tk) gene, was rendered dependent upon the expression of the human immunodeficiency virus type 1 (HIV1) tat and rev proteins. The SEAP or tk genes were placed between HIV1 splice donor and acceptor sites. One SEAP construct carried a series of alternating splice donor and acceptor sites. In all cases, the rev response element mapped within an intron. Despite such mimicry of the HIV1 genome, residual expression of the reporter gene in the absence of tat and rev was observed. These results, as well as non-specific T-cell recruitment, suggest limits to the specificity of using HIV-activated toxic gene expression to kill HIV-infected cells. PMID- 7481090 TI - Stochastic events in the amplification of HTLV-I integration sites by linker mediated PCR. AB - Human T-cell leukaemia virus type I (HTLV-I) proviral integration sites from an asymptomatic carrier and from the MT4 cell line were analysed by linker-mediated PCR (LMPCR) and inverse PCR (IPCR). LMPCR was more sensitive, allowing detection of a greater number of integrated proviruses. Reconstruction experiments using a cloned integrated HTLV-1 provirus indicated that > 100 copies were necessary to be detected frequently by LMPCR. To circumvent this problem, the LMPCR analysis was performed approximately 20 times per sample. Thus, for the MT4 cell line, the seven major integration sites were accompanied by approximately 20 clones of lesser frequency. For an asymptomatic HTLV-I carrier, nine integration sites were identified in a single amplfication, while a further 9 followed from 14 additional reactions. These findings show that there is a stochastic element to sampling HTLV-I integration sites by LMPCR, which tends to underestimate the actual number of HTLV-I bearing clones. Accordingly, those detected in at least two reactions represent the most abundant clones. PMID- 7481091 TI - Morphological changes in lymph nodes and expression of VCAM1 and cytokines at the late stages of SIV-induced disease in rhesus monkeys. AB - Four patterns of structural alterations were found in lymph nodes (LNs) from rhesus monkeys 17 to 34 months after infection with simian immunodeficiency virus (SIV-mac251). SIV p27gag antigen and viral particles were localized either between the processes of follicular dendritic cells (FDCs) or in the cytoplasm of macrophages. In hyperplastic follicles, enlarged germinal centres contained numerous Ki67+ proliferating centroblasts which were rather rare in light zones occupied by the CD23+ FDC network. Involuted follicles contained a small number of Ki67+ centroblasts and the CD23 labelling was limited to a very small apical zone. A correlation was found between the morphological characteristics of the follicles (hyperplasia-involution) and the level of expression of the vascular cell adhesion molecule 1 (VCAM1) on FDCs. A gradient in VCAM1 intensity with no expression in the subcapsular-intermediary sinuses, low membrane labelling in the mantle and strong expression in the FDC network was observed. IL1 alpha+ and IL6+ (interleukin) cells (lymphocytes and macrophages) were detected in the mantle, the interfollicular area and the medulla of LNs. Expression of the tumour necrosis factor alpha and ultrastructural markers of interferon alpha production were found in a few FDC and macrophages. Our findings indicate a close relationship between the morphofunctional properties of FDC and the LN structure in SIV infection. PMID- 7481092 TI - Broadly neutralizing, MN-like PND-directed antibodies in Rwandan children with long-term HIV1 infection. AB - Sera from 11 perinatally HIV1-infected Rwandan children with prolonged survival were tested in vitro for the presence of neutralizing antibodies against different HIV1 strains. These 11 sera from long survivor (LS) children were compared with 16 sera from Rwandan children with AIDS. Sera from HIV1-infected children exhibited the greatest neutralizing activity against HIV1MN cell-free infection. They also inhibited HIV1RII and HIV1LAI cell-free infection with lower titres. Higher neutralization titres were observed in sera from LS compared to the AIDS group, with a significant difference for HIV1MN and HIV1LAI strains. Sera from LS children also inhibited syncytium formation induced by HIV1MN infected cells with higher titres than AIDS children. Sera from the HIV1-infected children showed reactivity to the HIV1MN V3 peptide, as well as to both the US/European and the African consensus V3 peptides. Higher reactivity was observed in sera from LS than from AIDS children, and the difference was significant toward the African consensus peptide. The LS children also had significantly higher V3MN IgG avidity than the AIDS children. These data support the notion that the humoral response to the V3 domain, associated with a broadly neutralizing activity, may be an important factor in the prolonged survival of these children. The specificity against HIV1MN also suggests that an antigenically MN-related strain may be prevalent in Rwanda, and that an MN related principal neutralizing domain sequence could be an important determinant for candidate vaccines in this part of Africa. PMID- 7481093 TI - Inhibition of herpes simplex, rabies and rubella viruses by lectins with different specificities. AB - A series of eleven lectins were investigated for their inhibiting activity towards herpes simplex type 1, rabies and rubella viruses. Herpes simplex virus adsorption was inhibited by Limulus polyphemus (LPA), Canavalia ensiformis (ConA) and wheat germ (WGA) agglutinins, whereas Bauhinia purpurea (BPA) and soybean agglutinins were more effective if present during viral replication; rabies virus attachment to susceptible cells was prevented by LPA and Narcissus pseudonarcissus (NPA) agglutinin whereas WGA and BPA were inhibitory at a later step; rubella virus multiplication was affected only after the attachment step by NPA, ConA and WGA. PMID- 7481094 TI - Measurement of rabies virus N protein in rabies vaccines. AB - In this study, we designed a capture ELISA using a monoclonal antibody specific for the N protein, the major protein of the rabies virus nucleocapsid, to measure the N protein content in rabies vaccines. Free N protein content was compared in the two types of rabies vaccine currently used in humans: suckling mouse brain (SMB) vaccine prepared from rabies virus-infected brain tissue, and tissue culture (TC) vaccine prepared from supernatants of rabies virus-infected cells. It was found that SMB vaccines contained considerably higher amounts of N protein than most of the TC vaccines. Possible implications concerning the efficacy of rabies vaccines are discussed. PMID- 7481095 TI - Detection of IgM antibodies specific for measles virus by capture and indirect enzyme immunoassays. AB - During a measles outbreak, 112 serum specimens from 88 hospitalized patients were received in our laboratory for investigation of a morbilliform rash. These specimens (88 acute- and 24 convalescent-phase) were tested for the presence of measles-specific IgM antibodies by a capture EIA (enzyme immunoassay) using peroxidase-conjugated measles virus antigens and by an indirect EIA. Commercially available indirect EIA kits for measles-specific IgM antibodies were also used and compared with our homemade EIAs. Specificity studies included a collection of serum specimens containing rheumatoid factor, antinuclear antibodies or IgM antibodies specific to other viruses, and sera from blood donors and healthy children. Sensitivity of capture EIA and indirect EIA to detect measles IgM was 91.8 and 90.3%, respectively, and specificity was 98.2% for both tests. Specific IgM antibodies were detected in 70.5% of serum specimens at the first day after rash onset and were present for a month following the rash. Among the commercial measles IgM detection assays, EIA "Behring" was found to be a valid alternative for detection of measles virus-specific IgM. PMID- 7481096 TI - My response to "polio vaccines and the origin of AIDS". PMID- 7481097 TI - The determinant of severe cerebral dysfunction in patients undergoing emergency extracorporeal life support following cardiopulmonary resuscitation. AB - We investigated the factors associated with cerebral dysfunction in patients undergoing extracorporeal life support (ECLS) following conventional advanced cardiac life support (ACLS). The subjects were 9 patients in whom ECLS was started following ACLS because of intractable cardiac arrest. We investigated whether the irreversibility of cerebral dysfunction during ECLS was related to the cardiopulmonary resuscitation (CPR) time, arterial pH and blood gases, hemoglobin concentration (Hb), peak arterial pressure (PAP) before the start of ECLS and total doses of epinephrine and sodium bicarbonate administered during CPR. Two of the 3 patients who recovered consciousness were weaned from ECLS and survived, while all 6 patients who did not recover from coma were not weaned and died. There was no difference in the CPR time, Hb and PAP before the start of ECLS along with total doses of epinephrine and sodium bicarbonate administered during CPR between the patients who recovered consciousness and those who did not. In addition, there was no difference in arterial pH and blood gases except the arterial oxygen tension (PaO2) between the groups. The PaO2 values before the start of ECLS in the patients who remained in coma ranged from 34 to 58 mmHg, whereas those in the patients who recovered consciousness ranged from 132 to 442 mmHg. The PaO2 values before the start of ECLS in the patients who remained in coma were less than 60 mmHg, whereas those in the patients who recovered consciousness were over 60 mmHg. The present study suggests that hypoxemia during CPR may play a major role in severe cerebral dysfunction in patients undergoing ECLS and PaO2 during CPR. PMID- 7481098 TI - Failure of the laryngeal mask airway (LMA) to ventilate patients with severe tracheal stenosis. AB - The Laryngeal Mask Airway (LMA) was used during the resuscitation process in two patients with severe tracheal stenosis. In both cases the LMA failed to restore ventilation and both succumbed as a result of hypoxemic cardiac arrest. PMID- 7481099 TI - Post-resuscitation disease--a new nosological entity. Its reality and significance. AB - Experimental and clinical investigations of patients resuscitated after cardiac arrest or terminal states, testify to the fact that in the post-resuscitation period alongside processes of recovery and compensation, a number of new pathological phenomena develop. The latter differ substantially from those caused by ischemia and hypoxia. These post-resuscitation processes involve not only the CNS, but also the entire body and may lead to severe disability and even death of the seemingly successfully revived body. The data available suggests that this post-resuscitation pathology is a new nosological entity--a post-resuscitation disease. This disease has it own specific etiology, pathogenesis, variants of the clinical course (a number of syndromes) and the system of treatment and rehabilitation. In view of the ever wider use of resuscitation in clinical practice, it is expedient to organize an all-round study of this nosological entity, the optimum systems of its treatment, and to include this entity into International Classification of Diseases of WHO. PMID- 7481100 TI - The opioid/anti-opioid balance in shock: a new target for therapy in resuscitation. AB - Teleologically, pain is of paramount importance for survival and induces the organism to cope in an active way with aggressions from a basically hostile environment. While the activation of endogenous analgesic (opioid) systems typically occurs in conditions of surrender (pre-terminal conditions, sustained tortures, etc.), the activation of endogenous anti-analgesic systems triggers mechanisms of active or passive defence (such as camouflage) aimed at survival. The distinctive features of the main anti-analgesic systems (melanocortinergic, cholecystokininergic, thyroliberinergic) and the dramatic results obtained in experimental pre-terminal conditions (hemorrhagic shock, prolonged respiratory arrest) with the administration of their neuropeptide transmitters (ACTH and several ACTH-fragments, including alpha-MSH, CCK peptides and thyrotropin releasing hormone) are here reviewed. The study of the mechanisms underlying the resuscitating effects of these neuropeptides has led to the discovery of the (often extremely potent) resuscitating effect of other drugs (protoveratrines, nicotine, centrally-acting cholinergic agents, ganglion-stimulating drugs). It is particularly remarkable that in pre-terminal conditions these neuropeptides and drugs have highly impressive effects on cardiocirculatory parameters at doses that are almost or actually inactive under normal conditions, and that their resuscitating effect is obtained without the need for any other supportive treatment and at dose-levels well below toxic ranges. Finally, in hemorrhage shocked animals, the treatment with anti-analgesic neuropeptides shortly after bleeding considerably extends the time-limit for an effective and definitively curing blood reinfusion. This would be of self-evident importance in clinical practice, because an extremely simple, non-toxic first-aid treatment in the field, shortly after a massive hemorrhage, could resuscitate the patient for a period sufficient to effectively set up the most appropriate in-hospital treatment. PMID- 7481101 TI - Pilot study of intravenous magnesium sulfate in refractory cardiac arrest: safety data and recommendations for future studies. AB - Recent case reports have evidenced a temporal association between administration of i.v. magnesium sulfate (M) and resuscitation from prolonged cardiac arrest refractory to standard (S) ACLS attempts. However, speculation has arisen that M as a vasodilator, may decrease aortic diastolic and coronary perfusion pressure (CPP), aortic systolic and cerebral perfusion pressures (CePP), which may decrease resuscitation rates and neurologic recovery, as compared to standard ACLS alone (SA). OBJECTIVE: To resolve positive beginning evidence vs. negative theory, we conducted a pilot study of M+S vs. SA in refractory cardiac arrest on resuscitation rates (% R, return of stable pulses > 30 min without CPR, first in hospital cardiac arrest > 5-min duration) and neurologic recovery/survival to hospital discharge (SHD). METHODS: All patients from 1 January 1990-31 December 1991 at Rose Hospital, in cardiac arrest refractory to S through the first epinephrine dose (including 3 defibrillation attempts with pulseless VT/VF) were included in the data analysis, except: (1) patients with trauma, known poisoning, < 18 years, pregnancy excluded; (2) Standard ACLS alone patients with cardiac arrest < 5-min duration were not included in the SA comparison group, because the shortest cardiac arrest time before i.v. MgSO4 administration in the M+S group was 5 min. M+S (N = 29) and SA (N = 33) groups were also comparable on mean age (72-73 years) in this open-label prospective case-matched control group study. RESULTS: SHD rates were nearly equivalent between M+S (5.2%) and SA (4.5%). Complete or partial neurologic recovery, as best neurologic status post-R, occurred in 21% (6/29) M+S patients vs. 9% (3/33) SA (P = 0.17), even though cardiac arrest time on the study code call for resuscitated patients averaged shorter with SA (14.2 min) than M+S (19.8 min). M was frequently administered late in the resuscitation attempt--code call to M administration averaged 16.5 min (< 10 min in only 4/28 patients). A trend toward increased % R with M was evidenced: 21% (7/33) SA vs. 35% (10/29) M+S (P = 0.21). A temporal association between M administration and first return of spontaneous circulation (ROSC) was also documented in 4 of 10 M+S patients (pulseless electrical activity (3)/pulseless VT (1)), who had first ROSC/R occur within 0.5-2.25 min following first i.v. M bolus delivery, after 11-30 min (mean = 20 min) of continuous pulseless rhythm refractory to standard ACLS. All M+S resuscitations occurred within the dose range 2.5-5 g (i.v. push): 3/6 (50%) and 7/13 (54%) R occurred with 1-3 g and 4-5 g MgSO4, respectively (at least 11/13 patients had peripheral i.v. delivery with 4-5 g M). Analyzing post-ROSC hypotension proved important, as 50% of pts with first recorded systolic BP post-ROSC < 90 mmHg were resuscitated vs. 83% with > 90 mmHg (P = 0.10). A trend toward increased post-ROSC hypotension was evidenced with i.v. MgSO4: Recorded first or second systolic BP < 90 mmHg post-ROSC occurred in 66% of M+S vs. 42% of SA patients. All 3 M+S patients having a wide open i.v. levophed infusion as vasopressor support, started immediately post-ROSC/i.v. MgSO4 with systolic BP < 90 mmHg and continued at least 15 min (titrating to a systolic BP approximately 110 mmHg), had a temporal association between M delivery and R after 14-30 min of continuous pulselessness refractory to S. CONCLUSION: Human research determining whether i.v. MgSO4 increases long-term survival from refractory cardiac arrest should be vigorously pursued, as it is safe to proceed given the above described considerations. PMID- 7481102 TI - Hemodynamics and oxygen metabolism in the pig during long-term hypothermia: comparison of 2 pH strategies. AB - The aim of this study was to investigate the influence of acid-base management (pH stat or alpha stat) on hemodynamics and oxygen metabolism during long-term hypothermia in the pig. Seventeen female pigs were anesthetized, slowly cooled to 28 degrees C with cooling mats and kept at this temperature for 36 h. Thereafter, slow rewarming was performed with inhalation of a 40 degrees C warm air/oxygen mixture and insulation. Eight of the 17 pigs were ventilated according to the pH stat strategy and 9 according to the alpha stat strategy. Both groups were compared 4 times each for hemodynamics and metabolism during cooling, hypothermia and rewarming. The 2 strategies showed no significant difference in cardiac output, heart rate or mean arterial pressure. The only difference in hemodynamics was observed in mean pulmonary artery pressure and pulmonary artery resistance during hypothermia, showing higher values in pH stat animals. As for oxygen metabolism, oxygen consumption during hypothermia was significantly higher in alpha stat animals. Long-term hypothermia with spontaneous circulation revealed significant differences in oxygen metabolism and pulmonary artery pressure as well as resistance between alpha and pH stat acid-base management. These differences may be of importance when using moderate hypothermia in a clinical or experimental setting. PMID- 7481103 TI - Peritoneal cooling for mild cerebral hypothermia after cardiac arrest in dogs. AB - After normothermic cardiac arrest in dogs, we found that mild hypothermia (34 degrees C) of 1-2 h reduced brain damage, providing that hypothermia was achieved within 15 min of reperfusion. A clinically feasible rapid brain-cooling method is needed. As head-neck surface cooling alone in dogs was found to be too slow (0.1 degrees C/min), we reviewed peritoneal cooling in the Introduction and Discussion sections. PRELIMINARY EXPERIMENTS WITHOUT CARDIAC ARREST: In 5 dogs with spontaneous circulation and IPPV, 2 L of Ringer's solution at 10 degrees C were instilled into the peritoneal cavity, left for 5 min, and drained. Brain (tympanic membrane) temperature (Tty) decreased by a mean of 0.3 degrees C/min (12 min to 34 degrees C). Core (pulmonary artery) temperature (Tpa) decreased by a mean of 0.8 degrees C/min (5 min to 34 degrees C). COOLING AFTER CARDIAC ARREST: In our reproducible dog model of normothermic ventricular fibrillation cardiac arrest of 11 min (no flow), brief low-flow normothermic cardiopulmonary bypass (CPB) was used for reperfusion and restoration of spontaneous circulation (ROSC) within 2 min. In 24 dogs, mild hypothermia was induced by head-neck surface cooling with ice bags, starting with reperfusion, plus peritoneal lavage as above, starting with ROSC. All 24 dogs were resuscitated. Initial head-neck surface cooling alone over 2 min decreased Tty by only 0.15 degrees C/min. Subsequent additional peritoneal lavage decreased Tty by a mean of 0.3 degrees C/min (11 min to 34 degrees C); and Tpa 0.6 degrees C/min (7 min to 34 degrees C). There were no significant physiologic effects. We conclude that peritoneal instillation of cold Ringer's solution is more rapidly effective than other non intravascular cooling methods reported previously. Peritoneal cooling should be tried in patients during CPR. PMID- 7481104 TI - Post-ischemic hypothermia ameliorates ischemic brain damage but not post-ischemic audiogenic seizures in rats. AB - AIM: The objective of this study was to describe a pattern of recovery and histological nerve cell loss in Sprague-Dawley rats exposed to severe brain ischemia and to compare it to that of Wistar rats. METHODS: Ether- and ketamine anesthetized Sprague-Dawley rats were exposed to 3, 5, 6 and 10 min of normothermic severe brain ischemia (4 groups) induced by hypotension and neck compression. In group No. 5, the brain temperature was rapidly lowered, after 10 min of ischemia, to 30 degrees C during 45-50 min of recirculation. Wistar rats (group No. 6) served as controls (10-min normothermic ischemia). RESULTS: In Sprague-Dawley (S-D) rats, post-ischemic audiogenic seizures (PAS) increased with the duration of ischemia and reached 86% (6/7 rats), after 10 min of ischemia. Mortality rate was high (50% = 7/14 rats). No seizure activity was observed after 10 min of ischemia in 6 Wistar (W) rats, and all survived. In the S-D rats, 10 min of ischemia produced histopathological damage in all brain regions examined, except in the thalamus. Damage was less severe in the W rats. Post-ischemic hypothermia ameliorated hippocampal and cortical nerve cell damage, but had no effect on the incidence of PAS activity and mortality. In W rats, hippocampal nerve cell loss was much less severe than in the S-D rats and cortical damage was not observed. CONCLUSION: Sprague-Dawley rats develop post-ischemic audiogenic seizures more frequently than Wistar rats and mortality rate is high. The incidence of post-ischemic audiogenic seizures and mortality correlates well with the duration of ischemia. Post-ischemic moderate hypothermia (30 degrees C) significantly ameliorated the hippocampal and cortical nerve cell losses after 10 min of severe brain ischemia, but did not improve outcome. It appears that the high mortality rate of S-D rats following brain ischemia is related to frequent post-ischemic audiogenic seizures. PMID- 7481106 TI - Active transport in the alveolar epithelium of the adult lung: vestigial or vital? AB - Active secretion by mammalian fetal pulmonary alveolar epithelium is well recognized, as is the role of the adult epithelium in the secretion of surfactant. Recent studies have demonstrated active absorption by adult epithelium involving two sodium-dependent pathways. This finding has focused attention on how poorly we understand both the disposition of alveolar liquid and the physiological role of surfactant. In this paper we review the evidence that the adult mammalian alveolar epithelium absorbs solutes by active transport, and we assess the physiological importance of the resulting liquid movements. PMID- 7481105 TI - Role of endothelin-1 in the failure of cerebral circulation after complete global cerebral ischemia. AB - The role of endothelin-1 (ET-1) in the failure of cerebral circulation after cardiac arrest is unknown. We evaluated the effect of complete global cerebral ischemia that was induced on the plasma ET-1 concentration in 5 dogs, by occluding the ascending aorta and caval veins for 15 min. A fiberoptic catheter was inserted into the sagittal sinus for the continuous monitoring of venous oxygen saturation. Blood samples were collected before and at 0.5, 1, 2, 4, and 6 h after the ischemic insult. The ET-1 concentration in the sagittal sinus and abdominal aorta was assayed. Complete global cerebral ischemia resulted in a significant 3-fold increase in the sagittal sinus concentration of ET-1 (P < 0.01) that was associated with a significant decrease in the sagittal sinus venous oxygen saturation (P < 0.01); the arterial ET-1 concentration remained unchanged. The difference between arterial and venous ET-1 concentrations during the hypoperfusion phase subsequent to the ischemic insult may result from an increased secretion of ET-1 by the cerebrovasculature and a decrease in the clearance of ET-1 in the cerebral circulation. The result suggests that ET-1 may contribute to the failure of cerebral circulation after cardiac arrest. PMID- 7481107 TI - Hypoxia sensitive neurons in the caudal hypothalamus project to the periaqueductal gray. AB - Previous studies have demonstrated that the caudal hypothalamus modulates the respiratory responses to hypoxia and hypercapnia. In addition, many of the neurons in this area have a basal discharge related to the cardiac and/or respiratory cycles and are stimulated by hypoxia or hypercapnia. The purpose of the present study was to determine if these hypothalamic neurons project to a known cardiorespiratory area, the periaqueductal gray in the rat. In a first set of experiments, rhodamine-tagged microspheres were injected into the periaqueductal gray (PAG) to determine the areas of the caudal hypothalamus that project to the PAG. These studies revealed that the caudal hypothalamus sends strong ipsilateral and weak contralateral projections to the PAG. In a second set of experiments, single unit recordings were made from neurons in the caudal hypothalamus; the basal discharge of these neurons were examined with signal averaging techniques. Each neuron (n = 79) was tested for a response to inhalation of a hypoxic (10% O2) and a hypercapnic (5% CO2) gas. Antidromic activation techniques were then used to determine if neurons in the caudal hypothalamus send projections to or through the PAG. Nineteen percent (n = 15) of the hypothalamic neurons studied could be activated from the PAG; approximately 53% (n = 8) of these were excited by hypoxia and 27% (n = 4) by hypercapnia. Most of these neurons tested (42 of 64 neurons) had a basal discharge related temporally to the cardiac and/or respiratory cycles. These findings suggest that a caudal hypothalamic to periaqueductal gray projection is involved in the integrated response to hypoxia. PMID- 7481108 TI - Medullary neuronal activities in gasping induced by pharyngeal stimulation and hypoxia. AB - We examined the hypothesis that medullary respiratory-related and non-respiratory related neuronal activities are similarly altered with the "aspiration reflex", induced by mechanical stimulation of the epipharyngeal mucosa, and gasping, induced by severe hypoxia. Extracellular neuronal activities were recorded in decerebrate, paralyzed and ventilated cats. Phrenic activity and neuronal activities were monitored in eupnea and gasping. Seventy-one unit activities were recorded in the lateral medulla including the nucleus tractus solitorii (NTS), lateral tegmental field (LTF) and the nucleus ambiguus (NA). The respiratory modulation of a neuronal activity was quantified by a eta 2 statistic (Orem, J. and Dick, T., 1983, J. Neurophysiol. 50: 1098-1107). The eta 2 values of the units ranged from 0.02 to 0.93. Inspiratory-related activities with relative high eta 2 values (n = 16) were recorded in the region closed to the NTS. Phase spanning (n = 7) and expiratory-related activities (n = 10) were recorded in the ventral medullary region. Units with low eta 2 values (n = 29) and with no spontaneous activity (n = 9) in eupnea were recorded in the region of the LTF. In both "aspiration reflex" and gasping, inspiratory-related activities were augmented and expiratory-related activities were suppressed. Tonic units were activated and additional activities were recruited. The modulation of the neuronal activities to gasping induced by anoxia was identical to that induced by pharyngeal stimulation in either hyperoxia or severe hypoxia. We concluded that medullary gasping mechanism is recruited by pharyngeal stimulation.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7481109 TI - Circulatory and respiratory effects of an hypoxic stress in the Siberian sturgeon. AB - Changes in circulatory, ventilatory and acid-base variables were studied in Siberian sturgeon (Acipenser baeri) exposed to acute and severe hypoxia (PWO2 = 10 torr), followed by a rapid return to normoxia. This treatment caused a significant stress, revealed by the high levels of plasma catecholamines and cortisol. The moderate circulatory changes firstly observed would represent the effects of increased plasma catecholamine levels together with an increased adrenergic nervous tone on the cardiovascular system. Then, these effects were masked by a possible vagal reflex resulting in bradycardia. Deep hypoxia induced a ventilatory alkalosis combined with a moderate metabolic acidosis. The latter amplified concomitantly with a massive flush of lactate into the blood stream. The initial hyperventilation was followed by a deep ventilatory depression. During return to normoxia, hyperventilation resumed consistent with the repayment of an oxygen debt. Thus, the sturgeon, although considered as an archaic fish, developed the same adaptative responses as teleosts submitted to comparable hypoxic conditions. PMID- 7481111 TI - Hypoxic ventilatory responses in Tibetan residents of 4400 m compared with 3658 m. AB - Lifelong Tibetan residents of 3658 m ventilate as much and have hypoxic and hypercapnic ventilatory responsiveness as least as great as acclimatized newcomers, and likely greater than lifelong North or South American high-altitude residents. To determine whether Tibetans residing at altitudes > 3658 m maintained similar levels of ventilation, hypoxic and hypercapnic ventilatory responses, we transported 20 lifelong residents of > or = 4400 m to 3658 m for comparison with 27 similarly-aged male Tibetan residents of 3658 m. At 3658 m, the 4400 m compared with the 3658 m Tibetans had similar levels of minute ventilation and arterial O2 saturation, higher respiratory quotients but lower hypoxic ventilatory responses. We conclude that Tibetan residents of > or = 4400 m ventilate as much as Tibetan residents of 3658 m despite an altitude-associated blunting of their hypoxic ventilatory responses. Thus, factors other than hypoxic ventilatory chemosensitivity are likely to be important contributors to resting ventilation among Tibetan high altitude residents. PMID- 7481110 TI - Hypoxic ventilatory response predicts the extent of maximal breath-holds in man. AB - To understand the factors influencing breath-holding performance, we tested whether the hypoxic (HVR) and hypercapnic ventilatory responses (HCVR) were predictors of the extent of maximal breath-holds as measured by breath-hold duration, the lowest oxyhemoglobin saturation (SpO2min), lowest calculated PaO2 (PaO2min) and highest end-tidal PCO2 (PETCO2max) reached. Steady state isocapnic HVR and hyperoxic HCVR were measured in 17 human volunteers. Breath-holds were made at total lung capacity (TLC), at TLC following hyperventilation, at functional residual capacity, and at TLC with FIO2 = 0.15. SpO2 was measured continuously by pulse oximetry, and alveolar gas was measured at the end of breath-holds by mass spectrometry. PaO2min was calculated from SpO2min and PETCO2max. HVR was a significant predictor of both SpO2min and PaO2min. HVR and forced vital capacity were predictors of breath-hold duration by multiple linear regression. HCVR had no significant predictive value. We conclude that HVR, but not HCVR, is a significant predictor of breath-holding performance. PMID- 7481112 TI - Cardio-ventilatory control in rainbow trout: I. Pharmacology of branchial, oxygen sensitive chemoreceptors. AB - The effects of various neurochemicals on O2-sensitive chemoreceptor afferent discharge in the glossopharyngeal nerve (cranial nerve IX) were examined in an isolated, perfused first gill arch preparation from rainbow trout. Afferent neural activity from O2 receptors in the first gill arch increased in response to hypoxic perfusate and NaCN. Adrenergic agonists (epinephrine, norepinephrine and isoproterenol) had little effect on neural activity. Dopamine and 5 hydroxytryptamine (serotonin) caused a brief, small burst in chemoreceptor activity followed by a mild inhibition of receptor discharge. Acetylcholine and nicotine were potent neurochemical stimulants; muscarine had only a slight effect. While atropine completely blocked the effects of acetylcholine on receptor discharge, it only slightly inhibited responses to hypoxia and NaCN. Thus, although cholinergic mechanisms appear more likely than either adrenergic or serotonergic mechanisms to alter cardiovascular and ventilatory reflexes in fishes through their effects on O2-sensitive chemoreceptor activity, the transduction process involved in O2-chemoreception appears to be complex and not dependent on any single one of the neurochemicals tested. PMID- 7481113 TI - Dependency of upper airway patency on head position: the effect of muscle contraction. AB - In the present study we examined the effect of flexion and extension of the head on upper airway (UAW) patency in anesthetized dogs, and compared the dilatory and stabilizing effects of electrically stimulated UAW muscles at the different head positions. Flexion of the head increased UAW resistance (Ruaw) and reduced maximal flow (Vmax), but had little effect on the negative pressure at which UAW collapse occurred (Pcrit). Extension of the head, on the other hand, resulted in more negative Pcrit values and increased Vmax without significantly affecting Ruaw. Electrically induced UAW muscle contraction affected the pressure-flow curve and Ruaw, as well as Pcrit. Changing head position had a substantial effect on the dilatory and stabilizing effect of the various UAW muscles. However, independent of head position, genioglossus stimulation was most effective in reducing Ruaw and increasing Pcrit. We conclude that in the anesthetized, supine dog, head position affects the mechanical properties of the UAW and the effects of UAW muscle contraction. PMID- 7481114 TI - Dependence of collateral and small airway resistances of CO2 and volume in dog lobes. AB - We examined the hypothesis that collateral channels were identical within small airways and ducts, therefore both should respond similarly to chemical and mechanical stresses. A double lumen catheter was wedged into a segmental bronchus of the dog and humidified air or 10% CO2 in air (Vs) flowed at a segmental bronchial pressure (Pb) of 2 cm H2O. A small circular area about 1 cm diameter was peeled from the wedged segment and covered with a capsule glued to the surrounding pleura for measuring either a small airway flow (Vsaw) or capsule pressure (Pcap). Collateral resistance (Rcoll) and small airway resistance (Rsaw) were calculated as Rcoll = Pb/(Vs-Vsaw), Rsaw = (Pb-Pcap)/Vsaw, respectively. Hypocapnia (air) resulted in increases in Rcoll and Rsaw, while hypercapnia (10% CO2) generally had the opposite effect. Gcoll and Gsaw both increased linearly with lung volume (VL). The pattern of the responses of Rsaw closely paralleled those of Rcoll to local hypercapnia and hypocapnia, and to changing VL, implying that the major sites of resistance along collateral channels and along the airways are functionally and structurally similar. PMID- 7481115 TI - Hypoxic helium breathing does not reduce alveolar-arterial PO2 difference in the horse. AB - In a previous study we evaluated the mechanism of alveolar-arterial PO2 (AaPO2) reduction when nitrogen is replaced with helium in normoxia (FIO2 = 0.21). The reduction in AaPO2 was not due to changes in VA/Q inequality, pulmonary O2 diffusing capacity, or cardiac output, but to more complete diffusion equilibration as a consequence of the higher ventilation and thus PAO2 (which reduced the average slope of the hemoglobin O2 dissociation curve (ODC), and thus enhanced diffusive equilibration). We hypothesized that hypoxic He/O2 breathing in contrast would not reduce the AaPO2 because PAO2 and PaO2, although higher with He than N2, would remain constrained to the linear region of the ODC. Breathing hypoxic gas mixtures did constrain the PAO2 to the linear region of the ODC, even when PAO2 was increased by He/O2 breathing. Thus, the average slope of the ODC did not change when He replaced N2 and this explains the lack of change in AaPO2, as hypothesized. PMID- 7481116 TI - The effects of blood CO2 reaction rates on CO2 removal from muscle in exercised trout. AB - The purpose of this study was to examine the relative importance of some of the factors potentially limiting carbon dioxide (CO2) removal from the white muscle in the rainbow trout following exhaustive exercise. Inhibition of red blood cell carbonic anhydrase (CA) activity by acetazolamide (Az) infusion following exercise caused a large increase in both arterial and muscle PCO2 indicating retention of CO2 in the animal. Despite the increase in muscle PCO2, there was no significant decrease in muscle pH in Az-treated fish, probably due to the relatively high buffer capacity of white muscle. The presence of CA in the plasma had a transient, significant effect on blood PCO2 and pH, but no significant effect on muscle PCO2 or pH. Thus, CA treatment transiently enhances CO2 removal from the animal following exercise, but the extracellular catalysis of the CO2 reactions in the plasma does not appear to have a large impact on CO2 removal from the tissues. PMID- 7481117 TI - Magnitude and time course of acute hypoxic pulmonary vasoconstriction in man. AB - Acute hypoxic pulmonary vasoconstriction has an established role in the preservation of ventilation-perfusion balance. To further characterize this homeostatic response in man we have attempted to measure both the time course and magnitude of blood flow diversion from single hypoxic lobes. Lobar hypoxia (mean PO2 38 +/- 1.5(SEM)mmHg, mean PCO2 39.9 +/- 0.9 mmHg) was induced by inflating catheter-tip balloons in left upper lobe bronchi during fibreoptic bronchoscopy under local anaesthesia in 8 normal subjects. An index of lobar blood flow was obtained by acquiring dynamic scintigraphic lung images during a continuous intravenous infusion of the short-lived radioisotope krypton-81m dissolved in 5% glucose solution. In 3 subjects blood flow to the occluded lobes was monitored while the lobes were maintained under hyperoxic conditions (mean PO2 127.8 +/- 31.5 mmHg, mean PCO2 40.2 +/- 1.3 mmHg). Under hypoxic conditions the blood flow to the occluded lobes fell to 53% of baseline after 5 min with a mean time constant of 151 +/- 24.8 sec. Under hyperoxic conditions there was no significant change from baseline blood flow. We conclude that this technique has allowed us to monitor both the dynamic and steady state responses of the pulmonary circulation to lobar hypoxia in man. PMID- 7481118 TI - Embryogeny of oxygen consumption in 13 altricial and precocial birds. AB - The embryogeny of O2 consumption (VO2) was examined in 6 altricial and 7 precocial bird species (egg mass: 1.13 to 330 g). The 13 species studied were measured in most cases daily, some of them were continuously measured for 24 hours. 10 of 13 species exhibited a clear plateau-phase in about the last fourth of their embryogeny. Mean total energy production during incubation period was 1.83 kJ/g in altricial and 1.77 kJ/g in precocial species. No marked differences concerning the occurrence of a plateau phase could be found in the general course between the two developmental modes. This may reflect the existence of a basically identical course of the embryonic VO2 in the examined bird species. PMID- 7481119 TI - Qualitative course of embryonic O2 consumption in altricial and precocial birds. AB - The course of embryonic VO2 was examined in 24 altricial and 27 precocial bird species to test differences in its phenology. Some authors suggest that altricial species lack a plateau in VO2-development found in precocial species. Based on the recently available data we find the following: All species (except members of the family Megapodiidae) exhibit a plateau. Their VO2-development follows an identical basic course which can be described by an exponential increase interrupted by the plateau. There seems to be no fundamental difference between altricials and precocials with respect to the general presence of a plateau. In small altricials the plateau lasts only a few hours. It can be overlooked easily when the measurement is not continuous or when many individual measurements are averaged. However, the plateau-VO2 is lower (about 35%) and shorter (12.4% of incubation time; 16.9% in precocials) in the altricials. Total energy consumption during incubation is 1.9 kJ/g in altricial and 2.2 kJ/g in precocial species. PMID- 7481120 TI - [Spontaneous use of active and passive coping strategies for pain in patients with rheumatoid arthritis]. AB - The aim of the study was to determine association between active and passive pain coping strategies with some psychological and functional variables in 67 chronic RA patients. Active strategies reveal negative association with helplessness with as results of illness, and passive strategies reveal positive association with depression, helplessness and functional impairment. The patients who are using mostly active pain coping strategies reveal significantly less helplessness in coping with RA. PMID- 7481121 TI - [Diagnostic methods of detection of early degenerative changes in the lumbo coccygeal triangle in relation to the use of prostheses]. AB - In the evaluation of early degenerative changes of lumbocoxal triangle, that is of sacroiliac joints with the patients with above knee amputations, two diagnostic methods were used: X-ray method and scintigraphic method. X-ray method: pictures of sacroiliac joints according to Barsony as morphologic method show first degenerative changes in the period of six to twelve months after amputation. Scintigraphy of sacroiliac joints and the determination of s.i. index as functional method has established the degree of degenerative changes, that is their progression or stagnation. It has been seen that the degree of degenerative changes of sacroiliac joints in not the same with the patients who regularly use prostheses and those who do it occasionally. With the patients who use the prosthesis all the time, the degenerative changes stagnate. PMID- 7481122 TI - [Conservative treatment of the painful, stiff shoulder]. AB - The painful respectively stiff shoulder form an important part of orthopaedic practice. The treatment of patients with such a disease is at first always conservative except acute rupture of the tendon. Recurrences and those who do not respond to therapy require surgery. This study includes 86 patients treated for the painful shoulder. The were followed up and subjected to statistical analysis to determine the success of treatment by conservative methods. Conservative methods were in 83 shoulders, what amounts to a rate of 93 percent. Our results correspond to those reported in literature. PMID- 7481123 TI - [Measurement of motor strength of the hand using the Martin Vigorometer in evaluating the success of physical therapy in arthritis of the thumb]. AB - Motor strength of the fist was examined in 60 women with rhisarthrosis of the thumb before and after physical therapy with parafin compresses and magnetotherapy, as well as ultrasound through the water with VIGORIMETER "MARTIN". By measuring a better effect has been proved by using parafin compresses and magnetotherapy instead of the therapy with the ultrasound through the water. PMID- 7481124 TI - [Dynamometry of the hand--yesterday--today--tomorrow]. PMID- 7481125 TI - [Radiologic analysis of osseous changes in coxarthrosis]. AB - Morphologic and osteogenetic changes of the hip joint in patients with coxarthrosis were studied. Examination included 224 patients treated and operated on Department of Orthopaedics, Clinic Hospital Osijek. Coxarthrosis was proved in 331 hips, 183 cases (55,29%) referred to a secondary and 148 844,71%) to primary arthrosis. Osteogenetic changes on the head of femur were found in 148 x-rays (44,71%). The upper osteophyte of the head of femur was found in 79 x-rays of the hip (22,36%) but lower osteophyte of the head of femur only in 26 cases (7,85%). In 48 x-rays (14,50%) osteophytes were found both on the upper and lower sides of the head of femur. 217 x-rays of hips (65,56%) proved the existence of medial cervial osteophyte, according to frequency mostly type 1. Osteophyte of the roof of acetabulum were expressed on 158 x-rays (44,73). The double bottom of the acetabulum was studied on several measurement levels and showed from 0,24 to 2,35 mm observed from cranial to caudal measurement levels. The studies proved that the patients with coxarthrosis experienced a number of morphological and osteogenetic changes on the head and neck of femur as well as on the acetabulum. Their features on x-rays will depend on the cause and degree of the development of coxarthrosis. PMID- 7481126 TI - Pulmonary embolism: difficulties in the clinical diagnosis. AB - Pulmonary embolism is a common disease in the United States, affecting as many as 500,000 persons annually. Unfortunately, underdiagnosis of the disorder is also common, resulting in significant excess morbidity and mortality rates. Factors such as a lack of symptoms or the unavailability of effective testing techniques are not the reasons for pulmonary embolism's underdiagnosis. Instead, the disease is often often overlooked because the symptoms and signs caused by pulmonary embolism are nonspecific and may be confused with a variety of other cardiopulmonary disorders that have similar presentations. In addition to this overlap of symptoms and signs, pulmonary embolism frequently coexists with those diseases it mimics. This article describes the clinical characteristics of pulmonary venous thromboembolism, reviewing the symptoms and signs it typically exhibits, as well as the disease's routine laboratory test and chest radiographic abnormalities. These are contrasted with the clinical features of pneumonia, the disease for which pulmonary embolism is most commonly mistaken. I will show that clinical features can be used to increase suspicion for pulmonary embolism, but that additional diagnostic testing is necessary to definitively diagnose or exclude the condition. Accurate diagnostic tests are available for venous thromboembolism, even in the face of other coexistent cardiopulmonary disorders. As such, it should be possible to significantly reduce the morbidity and mortality rates caused by pulmonary embolism through both heightened clinical awareness and appropriate use of diagnostic testing strategies. PMID- 7481127 TI - Neoplastic mimics of pneumonia. AB - Several types of neoplastic conditions are included in the differential diagnosis of pneumonia. Bronchial obstruction with cancer can produce obstructive pneumonia that results in intractable infection. Bronchogenic carcinoma and metastatic cancer involving the airways may produce this clinical presentation. Bronchioloalveolar carcinoma is a relatively common form of primary lung cancer that characteristically presents as a chronic infiltrate associated with cough, hypoxemia, shortness of breath, and mucus hypersecretion. This cancer has two distinct histological types with markedly different prognosis. The mucinous variety is much more likely to be multicentric and rapidly progressive whereas the nonmucinous variety may be localized. Lymphoproliferative diseases may also present in an infiltrative appearance. Kaposi's sarcoma infiltrating the lungs, particularly associated with acquired immune deficiency syndrome, presents a diagnostic dilemma because of the high incidence of pulmonary infection in these patients. PMID- 7481128 TI - Radiation pneumonitis: a mimic of infectious pneumonitis. AB - The onset of fever and pulmonary infiltrates in patients who have been treated with thoracic irradiation is a relatively common occurrence. Radiation pneumonitis and infectious pneumonitis share many clinical features. The major objective of the clinician in evaluating patients who present with a febrile pneumonitis syndrome is to establish the correct diagnosis with as much certainty as possible. This article will review the pathogenesis, histopathology, and clinical features of radiation pneumonitis. Certain clinical and radiographic aspects of radiation pneumonitis will help the clinician arrive at the correct diagnosis. The patient with radiation pneumonitis will present with an insidious onset of dyspnea, fever, and nonproductive cough. Review of the chest radiograph at presentation and all chest radiographs since the completion of radiation therapy will provide the key to the clinical diagnosis of radiation pneumonitis. The infiltrate characteristically has a sharp margin that conforms to the port of irradiation. If the chest radiograph is not characteristic, it will be necessary to rule out infection. If noninvasive studies are nondiagnostic, then bronchoscopy may be necessary to exclude potential infectious agents. PMID- 7481129 TI - The fibroproliferative phase of late adult respiratory distress syndrome. AB - Tissue response to insults is similar regardless of the tissue involved, and occurs in two sequential and interconnected steps, inflammation and fibroproliferation. Adult respiratory distress syndrome (ARDS) is a disease characterized by acute onset of diffuse and severe inflammatory reaction of the lung parenchyma with loss of compartmentalization, resulting in protein rich exudative edema. Following tissue injury, a complex pattern of responses begins to repair the lung. Ineffective repair is evident histologically with extensive pulmonary fibroproliferation and clinically with fever (without a source of infection) and inability to improve lung function. We will review recent observations indicating that an exaggerated pulmonary inflammatory response plays a key role in the progression of ARDS. We will provide a unifying pathogenetic model of ARDS, showing how the evolution from acute to chronic inflammation explains the progression of histological, laboratory, clinical, and physiological findings seen during the course of unresolving ARDS. PMID- 7481130 TI - Pulmonary sarcoidosis: a mimic of respiratory infection. AB - Sarcoidosis is an idiopathic multisystem disorder with several clinical and roentgenographic features suggestive of respiratory infection. In the absence of infection, it is characterized by the microscopic presence of noncaseating epithelioid granuloma in affected tissues. When present, constitutional symptoms, fever, coughing, and exertional dyspnea usually develop insidiously, although occasionally Lofgren's syndrome--the triad of bilateral hilar adenopathy, erythema nodosum and polyarticular arthritis--may herald the onset of acute disease. Pulmonary involvement is the roentgenographic hallmark of sarcoidosis; bilateral hilar adenopathy is the most common manifestation. However, parenchymal infiltrates and pleural effusion may occur. Although numerous bacterial and fungal organisms may mimic the clinical and roentgenographic features of sarcoidosis, tuberculosis and fungal infections associated with granulomatous inflammation are the infectious processes most apt to cause diagnostic confusion. Several diagnostic clues are available to the clinician confronted with the consideration of sarcoidosis. Roentgenographic staging of the disorder (stage 0, normal radiograph; stage I, isolated bilateral hilar adenopathy; stage II, hilar adenopathy and parenchymal involvement; stage III, isolated parenchymal involvement; and stage IV, parenchymal fibrosis) provides a framework on which a differential diagnosis of likely infectious agents may be constructed and a history of travel to regions of endemic fungal infection may further narrow the differential diagnosis. An unexplained exudative lymphocytic pleural effusion or CD-4 lymphocyte predominance in bronchoalveolar lavage (BAL) fluid may also suggest a diagnosis of sarcoidosis. However, the definitive diagnosis of sarcoidosis is dependent upon the histological demonstration of noncaseating granuloma and the exclusion of infection in the appropriate clinical and roentgenographic setting. PMID- 7481131 TI - Neurologic aspects of hyperventilation syndrome. PMID- 7481132 TI - Panic attacks and panic disorder: the great neurologic imposters. AB - Patients who experience panic attacks, panic disorder, or agoraphobia have significant impairment associated with their disorders. The cost to society in health care costs as well as the human suffering and mortality is high and may be even higher than necessary because of misdiagnosis and inadequate treatment of these patients. Although we do not have many answers in the areas of pathophysiology or neurochemistry of panic disorder, we do have effective treatments for panic disorder and agoraphobia. If these are conceptualized as distinct disorders with specific symptoms, making a diagnosis of panic disorder or agoraphobia is relatively easy. Making the correct diagnosis may save the patient many months or years of suffering and many inappropriate tests or treatments. PMID- 7481133 TI - Funny spells in neuroendocrine disorders. PMID- 7481134 TI - Unusual seizure types. AB - There is no consensus as to what constitutes a diagnostic complex partial seizure type for any given focus. Nevertheless, some forms are seen infrequently, such as in the patient just described who had parietal seizures, whereas others, such as the bland onset followed by oral-alimentary automatisms of temporal lobe epilepsy, are more common. It helps to be aware of the possible origins of these unusual forms when differentiating electrical seizures from pseudoseizures, planning for use of specific pharmacologic treatments, or planning a surgical resection. Although one cannot recommend a surgical resection based on semiology alone, one can place intracranial electrodes more strategically when the hypothesis regarding the seizure focus includes the clinical characteristics of the patient's seizures. PMID- 7481135 TI - Autonomic epilepsy. PMID- 7481136 TI - Nonepileptic events. PMID- 7481137 TI - Migraine aura and prodrome. PMID- 7481138 TI - Transient global amnesia. PMID- 7481139 TI - Paroxysmal movement disorders. PMID- 7481140 TI - Sleep-related spells associated with parasomnias and narcolepsy. PMID- 7481141 TI - Paroxysmal nonepileptic disorders in children and adolescents. PMID- 7481142 TI - Undine's course. PMID- 7481143 TI - Comparison of vascular access devices. AB - OBJECTIVE: To provide an overview of venous access device designs and methods of insertion and removal. CONCLUSIONS: Venous access devices are indicated for many patients who require reliable long-term venous access. Three types of venous access devices are available including nontunneled, tunneled, and implanted ports. Since their introduction into clinical practice, the widespread use of these devices has had an enormous impact on cancer treatment by decreasing the overuse of peripheral veins while allowing for more flexibility and choice of the type of device used. Although numerous devices are available, each offers unique designs and performance expectations. Each type of device has similar features and can be used for intravenous drug and nutritional therapy, administration of blood products, and withdrawal of blood. IMPLICATIONS FOR NURSING PRACTICE: Even if only a single device is used in a given setting, the nurse must have a basic understanding of all types of venous access devices. It is imperative that the nurse be fully cognizant of the anatomic position and structure of the major vessels associated with the central venous system, especially for the insertion of peripheral central catheters. Understanding the venous system and venous access design can assist in preventing or assessing potential complications. PMID- 7481144 TI - Selection of vascular access devices and nursing care. AB - OBJECTIVE: To provide an overview of the patient selection criteria for a vascular access device (VAD) and its maintenance care. CONCLUSIONS: The availability of different types of devices had led to confusion on the part of many health care professionals as to which device is the best for their purpose. Selecting the appropriate VAD is an interdisciplinary decision based on patient characteristics, type of therapy, support system availability, and cost. Numerous controversial issues surround the care of VADs, and research results have not yet resolved these practice issues. Although various procedures are used throughout the country, it can be reasonably concluded that the fundamental requirement for successful maintenance of a VAD is strict adherence to established care protocols. IMPLICATIONS FOR NURSING PRACTICE: Assessment of the need for a VAD is an ongoing process. Nurses are in a perfect position to assess the factors that precede device selection, including the frequency of access, type of intravenous therapy, and the condition of the patient's peripheral veins. The vast array of devices demands specialized knowledge in selecting the appropriate VAD, providing correct care, and evaluating the device once inserted or implanted. PMID- 7481145 TI - Management of infection and occlusion associated with vascular access devices. AB - OBJECTIVE: To provide an overview of the etiology, assessment, diagnosis, and management of infections and occlusions that occur with venous access devices (VADs). CONCLUSIONS: The two major complications of VADs are infections and occlusions. Several strategies have been attempted to prevent and treat infections and occlusions; however, one specific method has yet to be determined. As a result, major controversies exist regarding the best method to manage these complications. Controlled randomized studies are needed to examine each type of VAD in relation to preventive and treatment strategies. IMPLICATIONS FOR NURSING PRACTICE: Preservation of VADs demands the development of and adherence to specific guidelines for maintenance care as well as an awareness of the incidence, presentation, and management of complications. Clearly, prevention is the key to effective management of infection and occlusion associated with VADs. Nurses must be familiar with institutional policies and procedures for the care of VADs. PMID- 7481146 TI - Rare complications of vascular access devices. AB - OBJECTIVE: To provide an overview of rare complications of vascular access devices in terms of etiology, assessment, management, and follow-up. CONCLUSIONS: Rare complications of vascular access devices are serious and in some cases life threatening. These complications include pinch-off and catheter fracture, catheter malposition and migration, cardiac perforation, extravasation, breakage, and defective devices. Knowledge and awareness of these complications can contribute to accurate identification and immediate management. Documentation and reporting of complications to the Food and Drug Administration helps to ensure the safety and efficacy of vascular access devices. IMPLICATIONS FOR NURSING PRACTICE: Developing the expertise needed to manage these types of complications is a continual challenge to the oncology nurse. Although managing rare complications is usually performed by a physician, recognizing the clinical manifestations will alert the nurse to seek medical advice immediately, thus decreasing the risk of mortality. All health care professionals are responsible for continually monitoring vascular access devices, consistently documenting findings, and communicating recommendations for care. PMID- 7481147 TI - Arterial, peritoneal, and intraventricular access devices. AB - OBJECTIVE: To provide an overview of access devices used to treat cancers through the arterial, peritoneal, and intraventricular body systems. CONCLUSIONS: Short term and long-term devices have been developed over the last 35 years for cancer treatment. Although less amenable to standard methods of therapy, the various access devices available to access the arterial, peritoneal, and intraventricular systems have provided a safe and reliable means for drug therapy. Access devices assist in delivering high concentrations of drugs directly to the center of the tumor. Complications and toxicities are inherent with these devices from the drug therapy as well as the device. Nursing assessment can provide early identification of potential problems and implementation of appropriate interventions. IMPLICATIONS FOR NURSING PRACTICE: As the availability of these devices increases, so must the nurse's knowledge base to provide optimal safe care. Oncology nurses are challenged to know the differences between the devices, the device of choice for the individual patient, insertion procedures, and maintenance protocols. PMID- 7481149 TI - Ambulatory infusion pumps: application to oncology. AB - OBJECTIVE: To provide an overview of the various types of ambulatory pumps available in relation to mechanism of delivery, patient selection criteria, potential complications, and educational support. CONCLUSIONS: Ambulatory pumps are an important means of drug delivery in a wide range of clinical conditions. Technologies permitting drug self-administration and automated drug delivery outside the hospital have become increasingly more sophisticated and adaptable; more than 600 models are available. With the shift of health care delivery from the hospital setting to the outpatient and home settings, reliable effective ambulatory pumps are necessary to safely deliver medications. IMPLICATIONS FOR NURSING PRACTICE: The use of ambulatory pumps is becoming more common as they are initiated as a part of front-line patient therapy. The nurse involved in the selection and management of ambulatory pumps is challenged to be familiar with the large variety available. Selecting the best type of pump is based on several factors, such as the type of therapy, disease state, reimbursement, and the ability to operate and troubleshoot complications. PMID- 7481148 TI - The use of access devices in cancer pain control. AB - OBJECTIVE: To provide an overview of access devices and infusion pumps available for pain management. CONCLUSIONS: Pain still remains a significant problem in patients with cancer. Access devices and infusion pumps have expanded pain management options for patients unable to use simple routes for medication administration. When access devices are selected and used appropriately, pain relief can be accomplished safely. IMPLICATIONS FOR NURSING PRACTICE: Nursing assessment is essential to develop an effective pain management regimen. Because patients are the best source of information about their pain, the nurse is in an excellent position to determine if the placement and use of an access device could be useful in controlling a patient's pain. PMID- 7481150 TI - Patient education and compliance issues associated with access devices. AB - OBJECTIVES: To explore the variables impacting patient education and to describe techniques for implementing successful education. CONCLUSIONS: Patients have the inherent right to be informed about all available options, significant risks, and treatment options. Throughout the teaching process, adequate time is needed for the patient to demonstrate skills in care and to voice concerns. Written literature and videos from the manufacturer or developed within the institution augment the nurse's teachings. Having a family member or significant other present is necessary to ensure that the teaching is complete. IMPLICATIONS FOR NURSING PRACTICE: Nurses are responsible for teaching patients about device care and how to troubleshoot complications. Nurses must be aware of factors that can affect learning, such as the patient's age, physical and emotional status, educational level, and current stress level. Failure to provide proper instruction may resort in the omission of vital steps, which may create problems with any type of device. PMID- 7481151 TI - [Medical care of elderly patients]. PMID- 7481152 TI - [Is the indomethacin test able to indicate the etiological diagnosis of isolated fever?]. AB - The aim of this study was to determine if evolution of fever after administration of indomethacin to febrile patients could separate those with fever of infectious origin from those with non-infectious origin. All patients with a rectal body temperature superior than 38 degrees C for at last 5 days and without any antibiotic or antipyretic therapy for more than 48 hours, were included in a 1 year prospective study. Each patient received one time 50 mg of indomethacin and rectal body temperature was obtained every 3 hours for 12 hours. Forty-five patients were included, 20 in group I (fever of infectious origin) and 25 in group II (fever of non-infectious origin). The mean age and mean initial body temperature were similar in the two groups. After administration of indomethacin, mean duration and mean amplitude of abatement of fever were similar in the two groups. Hypothermia was observed more frequently in group II (28%) than in group I (10%) (p < 0,05) and was preferentially associated to a malignant lymphoma. Also, diagnostic procedure with indomethacin is of little interest to separate fever of infectious origin from other, but hypothermia would suggest a fever due to a malignant lymphoma. PMID- 7481153 TI - [Sleep apnea and fibromyalgia: the absence of correlation does not indicate an exclusive central hypothesis]. AB - Fibromyalgia (FM) is a chronic painful syndrome characterized by widespread aching and points of tenderness, changed perception of pain and reduced brain serotonin. Abnormal EEG patterns have been reported in this condition. A study of FM occurrence in subjects with sleep abnormalities demonstrated by polysomnography was performed. Fifty patients (group 1:29 with sleep apnea and group 2:21 with poor sleep without sleep apnea) and 31 control subjects (without any sleep abnormalities) were submitted to the same clinical research of FM (ACR criteria). There was one 1 FM in group 1 (3.4%), one FM in group 2 (4.7%), and one FM in control group (3.2%). Sleep abnormalities and particularly sleep apnea are not significantly associated with FM. Other pathophysiological factors than central mechanisms are probably involved in the pathogenesis of FM. PMID- 7481155 TI - [Myocardial infarction and revascularization. Current indications]. AB - The physiopathologic role of thrombosis in the genesis of myocardial infarction, began to be suspected early in the 20th century but its logical treatment, thrombolysis, was first used on a large scale only ten years ago. Today, it is well established that short, middle and long-term mortality is correlated to coronary permeability, the delay in the revascularization treatment start-up, its efficacy, its swiftness of action, and to the maintaining of permeability following reperfusion. The importance of time elapse before reperfusion is obtained was demonstrated as early as 1986 by the GISSI study. According to this study, the administration of streptokinase (compared to a conventional treatment) reduced mortality at 21 days respectively by 47%, 23%, and 17%, depending on whether patients were treated within one hour, three hours, or between 3 and 6 hours following the onset of the painful symptoms. One of the major teachings of the GUSTO study, reported at the end of 1993, was the confirmation of the so called "open artery" theory: mortality at 30 days was of 4.5% among patients whose coronary circulation was restored at the 90th minute, whatever thrombolytic treatment was used, compared to 8.9% when the coronary artery remained occluded. The value of aspirin in preserving coronary permeability following thrombolysis was demonstrated by the ISIS-2 study: mortality at 5 weeks was reduced by 23% in the group of patients randomised to receive only aspirin, while it was reduced by 25% in the group of patients randomised to be treated with streptokinase, and by 42% in the group randomised to receive both aspirin and streptokinase, compared to the group who received neither aspirin nor streptokinase. However, mortality during the first days following randomisation was identical among the groups, with or without aspirin, which suggested its action was rather one of prevention against reocclusion than one of accelerating dissolution of the thrombus. However, in spite of improved therapeutical protocols, a normal flow, which is the major criteria for a reduced mortality, is only obtained at the 90th minute in 54% of the patients who were administered the up-to-date treatment ie aspirin accelerated t-PA-heparin in combination.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 7481154 TI - [Skin manifestations related to hepatitis C viruses]. AB - The hepatitis C virus causes both hepatic and extrahepatic disorders, particularly as regards dermatology. The link between essential mixed cryoglobulinemias and the C virus infection has been clearly evidenced., whereas its frequency seems low in other systemic vasculitis such as polyarteritis nodosa. Similarly, the link between C virus hepatopathy and porphyria cutanea tarda is now proven. Lichen planus is also described as being associated with this virus, but further epidemiological studies are required to determine the exact prevalence of lichen in C virus hepatopathy cases. Finally, various cutaneous disorders, such as urticaria, erythema multiforme, dermo-hypodermitis, etc, occasionally arise during acute or chronic hepatitis C. PMID- 7481156 TI - [Systemic lupus erythematosus and Salmonella enteritidis osteomyelitis]. AB - Infections in systemic lupus erythematosus are frequent. However, osteoarticular Salmonella infections are rarely reported. We report a case of systemic lupus erythematosus diagnosed in a 15 year-old girl. Seven months later, she presented with fever and a localized collection of the upper extremity of the left tibia related to a Salmonella enteritidis acute osteomyelitis (sub periosteal abscess). The out-come was chronic and led to death. The authors emphasize the severity of non typhoidal salmonellosis in systemic lupus erythematosus. PMID- 7481157 TI - [Uveitis: late complication of undiagnosed Whipple disease]. AB - We report a case of severe chronic bilateral uveitis in a 60-year-old white man without neurological manifestations. Spondylarthritis and isolated malabsorption suggested the diagnosis of Whipple's disease. A normal duodenal biopsy delated the diagnosis, who was secondarily demonstrated an adenopathy biopsy and macrophage study on vitrectomy sample (macrophage distended by periodic acid Schiff positive material). With 1 year follow-up the uveitis was rapidly and completely cured by an antibiotic association (rifampicin and trimethoprim sulfamethoxazol). The characteristics of ocular involvement in Whipple's disease and treatment of the disease are discussed. PMID- 7481159 TI - [Principles of organization of a center for pain evaluation and treatment]. AB - Since the development during the sixties of the pioneer pain clinics in the United-States, the need of a pluridisciplinary approach of the chronic pain patient has progressively compelled recognition. The principles of organization of this new care units--the pin center--are now clearly determined. It has become classic to compare acute pain as a warning symptom with chronic pain as an illness in itself with its constellation of psychosocial factors. Chronic pain refers to a daily pain persisting for 3 to 6 months. Neurophysiological, neuropsychological and behavioral differences legitimate the acute/chronic distinction. We will consider the following items: the types of patient, the multidisciplinary model, the team functioning, the initial consultation, the multidisciplinary synthesis discussion, the somatician role, the psychiatrist role and the possible disadvantages. Beside the care mission, multidisciplinary pain centers also have a key role in clinical research and teaching. PMID- 7481158 TI - [Nephropathies caused by interferon alpha: apropos of 2 cases]. AB - Two cases reports of interferon alpha-associated nephropathy are reported. The first observation is a membranoproliferative glomerulonephritis and the second a renal microangiopathy. The different cases in the literature are reviewed and the pathophysiology is discussed. PMID- 7481160 TI - [Yersinia enterocolitica septicemia, iron overload and deferoxamine]. AB - Infections due to Yersinia enterocolitica are usually limited to the bowel. When infection is generalized, the role of iron overload and iron chelation has been discussed. We report the case of a 55 year-old patient with sideroblastic anemia who received repetitive transfusions and deferoxamine for 4 years and heme arginate for 2 months, and who was admitted in our institution for Yersinia enterocolitica sepsis. Treatment by third-generation cephalosporins and aminoglycosides has allowed favorable outcome. PMID- 7481161 TI - [Strategy of treatment of cancer pain in children]. AB - Most children with cancer will experience pain during their illness, whether it may be cured or not. They may suffer from acute pain related to treatments or to invasive procedures, or from prolonged pain due to the evolution of cancer or sequellae of treatment. Pain must be considered as a major symptom and must be suspected, diagnosed and evaluated. Physicians have to analyse the different components, the physiopathologic data, and causes of pain. Treatments have to be prescribed, adjusted to each patient and monitored in conformity with rigorous guidelines in order to obtain the best analgesic efficacy and the lowest side effect levels. The use of opioids is frequent and their doses are higher as those used in adults. Neuropathic pain is frequent in children suffering from cancer and requires treatment by means of antidepressant drugs if clinical signs of neuropathic pain are predominant. PMID- 7481162 TI - [Towards a vaccination against malaria]. AB - For 20 years, the prospect of anti-malarial vaccination has aroused many hopes, but in the end, it has mostly given rise to doubts and disappointment. If most attempts have been to no avail, this is because the issue at stake is amazingly difficult. Besides the very complex antigenic structure of the protozoa Plasmodium, there is first the existence of at least three different targets during the plasmodial cycle, then the necessity of appropriate adjuvants and, most of all, the imperfection of our experimental models. Recently, Pattaroyo and the various groups who worked with him have eventually met success with vaccine trials in man: they used a synthetic antigene, SPf66, on volunteers in South America, then on a larger population sample in East Africa. The results are still quite modest: people are protected against the malarial disease but not against the parasitemia and only in approximately 40% of cases. Nevertheless, these results have the merit of representing the first successful anti-malarial vaccination in man. Although great advances are still needed, a decisive step forward has been taken. Other types of vaccine will soon be tested by other groups (anti-gametocyte vaccines) and prospects of significant improvements are offered by the technique of DNA-vaccines. If it is now certain that one or several vaccines will be available in a near future, no one is able to set the time delay necessary to reach this stage. In any case, hoping that this type of vaccine will eradicate the disease is not realistic since a disease as complex as malaria, in terms of epidemiology, cannot be eliminated by only one method.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7481163 TI - [Sarcoidosic dactylitis]. PMID- 7481164 TI - [Apyrexia induced by non-steroidal anti-inflammatory agents in 5 cases of Hodgkin disease]. PMID- 7481165 TI - [Cardiac amyloidosis disclosed by abdominal arterial embolism]. PMID- 7481166 TI - On prevalence and incidence. PMID- 7481167 TI - Psychological reactions to information about risk of ischaemic heart disease in general practice. AB - OBJECTIVE: To examine the psychological reactions in 40-49 year old men diagnosed as having an increased risk for the development of ischaemic heart disease at a health examination in general practice. DESIGN: A multipractice study including a questionnaire about the psychological well-being before and 6 months after a health examination aimed at finding an increased risk for ischaemic heart disease. SETTING: General practice in the county of Aarhus, Denmark. Sixty five general practitioners. PARTICIPANTS: 123 men with and 150 men without an increased risk of ischaemic heart disease. OUTCOME MEASURES: Psychological well being was measured by the General Health Questionnaire (12 item version). RESULTS: No significant change in GHQ-scores after the screening examination. CONCLUSION: Information about increased risk of IHD in 40-49 year old men at a health examination in general practice did not change the psychological well being as measured by a General Health Questionnaire 6 months after the examination. PMID- 7481168 TI - General practitioners' management of female urinary incontinence. Medical records do not reflect patients' recall. AB - OBJECTIVES: To investigate the prevailing management of female urinary incontinence by Norwegian general practitioners (GPs), and to compare information given by the patients with information in their medical records. DESIGN: Incontinent women who had received reimbursement for incontinence aids or drugs were identified by the local Insurance Offices, and a random sample was interviewed about what examinations and treatments they had received. Similar information was later collected from their GPs. SETTING: The Bergen area, Norway. SUBJECTS: Forty-eight GPs and 82 patients. MAIN OUTCOME MEASURES: Actions taken according to the patients and their medical records. Kappa statistics were used to evaluate the strength of agreement. RESULTS: The GPs had done a gynaecological examination in 54%, a leakage provocation test in 12%, and a urinalysis in 73% of the patients. Thirty-two per cent of the women had been instructed in pelvic floor exercises and 13% in bladder training. The GPs had prescribed oestrogens for 48% of the older women (> 59 years), anticholinergics for 29% of those with urge or mixed incontinence. Overall Kappa between the two data sources was 0.37. CONCLUSION: GPs' incontinence management can be improved, especially regarding gynaecological examination, pelvic floor exercises, and the prescription of oestrogens. Retrospective chart data or patient interviews are unreliable data sources. PMID- 7481169 TI - Medical audit used for estimation of optimum level of outpatient care. AB - OBJECTIVE: To create a model for determining the optimum level of outpatient medical care and to determine this level. DESIGN: Expert committees were established comprising one or two hospital physicians and two general practitioners who checked medical records for all outpatient visits to doctors made by a defined population during a defined period of time. The determination of optimum level was made blindly by each member of the expert committee. SETTING AND PARTICIPANTS: The study comprised all visits to physicians during 10 weeks, in total more than 4,000, made by the people in a Swedish community, of which 2,084 were randomized. RESULTS: Total agreement between the members of the committee was initially reached for 84% of the visits and, after a common discussion between the committee members, for 99%. A general practitioner was considered to be the optimum level of care for 76% of the patients in the total series, in the upper ages (above 80) for about 85%. CONCLUSION: This method seemed suitable for determining the optimum level of care in a population and may be of value when planning for an optimum health care service. Based on the results from our study it seems reasonable to assume that general practitioners whose training corresponds to that of Swedish ones are competent to treat about 75% of all consultations. PMID- 7481170 TI - Psychosocial factors and heavy smoking during pregnancy among parous Scandinavian women. AB - OBJECTIVE: To analyse how psychosocial and social factors are associated with heavy smoking in the third trimester of pregnancy. DESIGN: Data were collected from a prospective study primarily initiated to study fetal growth retardation. (NICHD Study of successive small for gestational age births). SETTING: Uppsala in Sweden, Bergen and Trondheim in Norway. SUBJECTS: Study subjects were 775 para I and II, referred from GPs or maternity wards to the University Hospitals in Uppsala, Bergen and Trondheim. All women smoked at the time of conception. "Heavy smokers" were defined as women who smoked 15 or more cigarettes per day during the third trimester. MAIN OUTCOME MEASURE: Psychosocial factors and potentially associated heavy smoking in the third trimester. RESULTS: Young women and women without a partner smoked heavily more often than older women and married/cohabitant women. Growing up with just one parent and the experience of a difficult childhood were also significantly associated with heavy smoking before delivery. If the woman's family (except partner) did not approve of the pregnancy, the woman was more often a heavy smoker. The women who smoked heavily in the third trimester had started smoking at an earlier age than the rest of the smokers. CONCLUSION: The pregnant woman's previous and present psychosocial conditions are related to her heavy smoking in the third trimester of pregnancy. PMID- 7481172 TI - General practice consultations in central and northern Finland. AB - OBJECTIVE: To examine general practice consultations in health centres in central and northern parts of Finland. DESIGN: A questionnaire concerning all general practice consultations during one week. SETTING: Central and northern parts of Finland in a region comprising 72% of the area of the country and one-third of its population. PARTICIPANTS: 851 health centre doctors. OUTCOME MEASURES: Consultation rates by age and sex of the patients and by characteristics of doctors and their practices. RESULTS: The average consultation rate per 1,000 female inhabitants was 43 and per 1,000 male inhabitants 34 per week. For total contacts, a J-shaped association with increasing age was demonstrated for both sexes. The average number of consultations per doctor in a week was 72. Consultation rates were lowest among physicians over 45 years of age. A low consultation rate was also found among general practitioners not participating in the personal doctor programme and among those who worked in municipalities with over 30,000 inhabitants. CONCLUSION: In Finland, consultation rates per 1,000 inhabitants, as well as per general practitioner, are remarkably lower than in most European countries. The personal doctor programme with defined lists of patients seems to be associated with high consultation rates. PMID- 7481173 TI - Care for demented patients in different living conditions. Effects on cognitive function, ADL-capacity and behaviour. AB - OBJECTIVE: To assess if and how demented patients' living conditions affect their level of functioning. DESIGN: A prospective concurrent control-study. SETTING: Sundsvall, an industrial community in the middle of Sweden. PARTICIPANTS: Forty six demented patients in group-living (GL-group) and 62 controls (C-group). MAIN OUTCOME MEASURE: Cognitive function, ADL-capacity and behaviour were measured every third month during one year. RESULTS: Both groups deteriorated, but the decline was significantly slower in the GL-group regarding some ADL-functions, e.g. dressing and motor functions, whereas some behavioural disturbances were significantly more frequent in the GL-group. No differences were observed regarding the level of cognitive function. CONCLUSION: The living condition has small effects on cognitive function and ADL-capacity. The behavioural disturbances observed in GL should be the focus of future research. PMID- 7481171 TI - Assessment of sickness certification and concepts of musculoskeletal disease and illness in the general population. AB - OBJECTIVE: To investigate how laymen assess the need for sickness certification and how they use the concepts "disease" and "illness" in relation to musculoskeletal disorders. DESIGN: Mail questionnaire with simulated case histories. SUBJECTS: A stratified, random sample of 600 Norwegians, aged 16-69 years. MAIN OUTCOME MEASURES: Scores that measured the respondents' use of the concept "illness", and their view on need for sickness certification were constructed and analysed. RESULTS: Response rate was 54%. The respondents perceived seven case histories describing musculoskeletal health problems more often as "illness" (61%) than as "disease" (46%). There was a close correlation between the use of "illness" and assessed need for sickness certification. However, sickness certification could be suggested for cases with neck pain, even if the patient was not perceived as ill. Women suggested sickness certification significantly more often than men. Age, length of education, and personal experience with serious musculoskeletal health problems influenced suggestions on certification among men. However, these variables could not explain any difference in use of the concept "illness". PMID- 7481174 TI - Health and health care utilization among early retirement pensioners with musculoskeletal disorders. AB - OBJECTIVE: To estimate subjective health status among early retired individuals pensioned due to disorders of the musculoskeletal system. DESIGN: A survey of self-reported health status and health care utilization by means of a mailed questionnaire. SETTING: Early retirees (cases) and a random sample (controls) of individuals from the municipality of Kristianstad, Sweden. SUBJECTS: CASES: all individuals aged 25-59 years granted a full-time early retirement pension during the period 1986-1990 due to disorders of the musculoskeletal system (n = 450). The response rate was 83.6%. CONTROLS: An age- and sex-matched sample (n = 450). The response rate was 82.7%. MAIN OUTCOME MEASURES: Self-reported health status and health care utilization. RESULTS: Early retirement entailed a deterioration in self-reported health status among men, all ages, and women aged 25-54 years. Female retirees aged 55-64 reported an improvement in health status since retirement. Early retired female immigrants were less satisfied with their health status than the Swedish ones. Early retirees reported higher health care utilization than controls, but with a reduction since retirement. Drug consumption was high. CONCLUSION: The beneficial effects of early retirement were evident among old women. Men and young and middle-aged women may need support to adjust to life as a retiree. Special attention should be given to the female immigrant retiree. PMID- 7481175 TI - Decentralized diagnostics of beta haemolytic streptococci group A--introduction of a developmental model for microbiological consultation in primary health care. AB - OBJECTIVE: To define and evaluate forms for introduction of decentralized diagnosis in primary care. DESIGN: The study was divided into three phases. Intervention I. An intensive course on microbiological diagnostics in cases of tonsillitis with information about rapid EIA-based test kits for beta haemolytic streptococci group A. Intervention II. External quality assurance of the decentralized test. Evaluation. Recording of changes in work practice and attitude regarding decentralized diagnosis by means of a questionnaire. PARTICIPANTS: Thirty-three GPs and 36 laboratory technicians from 37 primary health care centres (PHC) in the county of Ostergotland, Sweden. RESULTS: Thirty one of the responding GPs (94%) considered that the information gained from quality assurance was useful. Twenty-nine GPs (88%) reported that they had changed their clinical practice to some extent, and 26 of the responding laboratory technicians (72%) reported that they had changed method after the interventions. CONCLUSION: Uniform acceptance was not achieved, but a large number of GPs and PHC laboratory technicians did conform to a rational-empirical strategy for change in clinical practice. PMID- 7481176 TI - Prevalence of diabetes mellitus and impaired glucose tolerance in a middle-aged Finnish population. AB - OBJECTIVE: To describe the prevalence of previously diagnosed and undiagnosed diabetes and impaired glucose tolerance (IGT) by sex, marital status, employment, exercise behaviour, and body mass index (BMI), and to compare previously diagnosed and undiagnosed diabetic patients. DESIGN: A survey of the total 55 year-old population living in a Finnish city on 1 October 1990. All the participants who were not on antidiabetic medication were invited for an oral glucose tolerance test (OGTT). SETTING: Oulu, a city in northern Finland. SUBJECTS: 1,008 eligible subjects (456 men and 552 women), of whom 345 men (76%) and 435 women (79%) participated. MAIN RESULTS: The prevalence of previously diagnosed diabetes was 6.1% in the men and 3.0% in the women and that of previously undiagnosed diabetes 4.5% of the men and 3.7% of the women. The prevalence of IGT was 28.6% in the men and 26.7% in the women. Diabetes was related to BMI, and previously undiagnosed diabetes was related to the amount of physical exercise in both sexes. One third of the undiagnosed diabetic men and half of the undiagnosed diabetic women were identified among the persons with marked obesity (BMI 30 kg/m2 or more). The group of previously diagnosed diabetic patients included a significantly greater proportion of retired people and people on sick leave than the group of undiagnosed diabetic persons. CONCLUSIONS: Diabetes is common in middle-aged Finns. Undiagnosed diabetes was detected especially in persons with marked obesity who were not physically active. PMID- 7481178 TI - Validity of self-reported smoking habits. PMID- 7481179 TI - Acute low back pain: is the number of patients taking sick leave exponentially related to time? PMID- 7481177 TI - Prevalence of known and previously unknown diabetes mellitus and impaired glucose tolerance in an adult Norwegian population. Indications of an increasing diabetes prevalence. The Nord-Trondelag Diabetes Study. AB - OBJECTIVE: To establish the prevalence of known and undiagnosed diabetes mellitus and impaired glucose tolerance in a representative Norwegian adult population according to the 1980 World Health Organization Expert Committee diagnostic criteria. DESIGN: Screening survey applying questionnaire and non-fasting blood glucose followed by a fasting and an oral glucose tolerance test. SETTING: The county of Nord-Trondelag, Norway, 1984-86. SUBJECTS: All inhabitants aged > or = 20 years (85,100); 90.3% participated. For previously unknown diabetes: all inhabitants > or = 40 years (53,001)--participation rate 84.7%. MAIN OUTCOME MEASURE: Prevalence. RESULTS: The prevalence of previously known diabetes was 2.6% in men and 3.2% in women. Total diabetes prevalence > or = 40 years was 4.8% in men and 5.6% in women. The prevalence increased continuously with age until 90 years. One patient in five was previously undiagnosed. Impaired glucose tolerance in those with an abnormal non-fasting value was rare; only 0.9% in men and 0.2% in women. The prevalence of known diabetes was three times higher than 20-30 years ago and four times higher than 50 years ago. Except for known diabetes in the very old the prevalence corresponded well with recent Nordic studies. IMPLICATIONS: The present diabetes prevalence was so high that interested general practitioners will get sufficient experience in follow-up of diabetic patients to facilitate a good quality of the care. PMID- 7481180 TI - Model selection techniques for the covariance matrix for incomplete longitudinal data. AB - In longitudinal studies with incomplete data, where the number of time points can become numerous, it is often advantageous to model the covariance matrix. We describe several covariance models (for example, mixed models, compound symmetry, AR(1)-type models, and combination models) that offer parsimonious alternatives to unstructured sigma. We evaluate each covariance model with longitudinal data concerning cholesterol as the repeated outcome measure. We discuss strategies for deciding the 'best' model and show a graphical technique for judging goodness-of fit of covariance models. PMID- 7481181 TI - Calculation of unconditional and conditional reference intervals for foetal size and growth from longitudinal measurements. AB - Methods for estimating unconditional and conditional reference intervals for foetal size and growth based on longitudinal observations are presented. The methods are based on simple random-effects regression models and involve transforming both the response and the covariate (timepoint). A dataset from a designed longitudinal study of foetal size is analysed in detail as a motivating example. PMID- 7481185 TI - Confidence limits for the population prevalence rate based on the negative binomial distribution. AB - This paper shows that the extension of the simple procedure of George and Elston in calculation of confidence limits for the underlying prevalence rate to accommodate any finite number of cases in inverse sampling is straightforward. To appreciate the fact that the length of the confidence interval calculated on the basis of the first single case may be too wide for general utility, I include a quantiative discussion on the effect due to an increase in the number of cases requested in the sample on the expected length of confidence intervals. To facilitate further the application of the results presented in this paper, I present a table that summarizes in a variety of situations the minimum required number of cases for the ratio of the expected length of a confidence interval relative to the underlying prevalence rate to be less than or equal to a given value. I also include a discussion on the relation between Clemans's confidence limits on the expected number of trials before the failure of a given device and those presented here. PMID- 7481182 TI - Estimating the reliability of an exposure variable in the presence of confounders. AB - In this paper we discuss estimation of the reliability of an exposure variable in the presence of confounders measured without error. We give an explicit formula that shows how the exposure becomes less reliable as the degree of correlation between the exposure and confounders increases. We also discuss biases in the corresponding logistic regression estimates and methods for correction. Data from a matched case-control study of hormone levels and risk of breast cancer are used to illustrate the methods. PMID- 7481184 TI - A hybrid paired and unpaired analysis for the comparison of proportions. AB - A method is presented for making inferences about the difference between two binomial proportions when there is partial overlap of the two samples. The method will simplify to the usual cases of two independent samples or paired samples (McNemar's method) depending on whether there is no or complete overlap. The method is used to compare sensitivities, specificities and Youden indices from a comparative study of trauma triage (screening) instruments. PMID- 7481183 TI - A comparison of frailty models for multivariate survival data. AB - This paper reviews some of the main approaches to the analysis of multivariate censored survival data. Such data typically have correlated failure times. The correlation can be a consequence of the observational design, for example with clustered sampling and matching, or it can be a focus of interest as in genetic studies, longitudinal studies of recurrent events and other studies involving multiple measurements. We assume that the correlation between the failure or survival times can be accounted for by fixed or random frailty effects. We then compare the performance of conditional and mixture likelihood approaches to estimating models with these frailty effects in censored bivariate survival data. We find that the mixture methods are surprisingly robust to misspecification of the frailty distribution. The paper also contains an illustrative example on the times to onset of chest pain brought on by three endurance exercise tests during a drug treatment trial of heart patients. PMID- 7481186 TI - Investigating a sequence of randomized phase II trials to discover promising treatments. AB - We consider clinical trial strategies to study diseases in which there is rapidly developing technology. We assume the availability of a limited number of patients for screening treatments over a time horizon, and that availability of new treatments for test is staggered over time. We assume further that patient response is binary and rapidly observable. We consider the strategy of conducting a sequence of two-armed randomized clinical trials. We carry over the treatment with the larger number of observed successes on the current trial to the next trial for comparison with a new treatment, with this process repeated at each step. For a fixed total number of patients (N), the number of trials one may conduct in sequence (k) is inversely related to the sample size per trial (2n), N = 2nk. We investigate how k and n influence (a) the expected success probability for the treatment selected at the end, and (b) the expected number of total successes for the N patients. The ultimate objective is to select one treatment, the winner at stage k, to test against a standard regimen in a randomized comparative phase III trial. PMID- 7481187 TI - Breaking the matches in a paired t-test for community interventions when the number of pairs is small. AB - There is considerable interest in community interventions for health promotion, where the community is the experimental unit. Because such interventions are expensive, the number of experimental units (communities) is usually small. Because of the small number of communities involved, investigators often match treatment and control communities on demographic variables before randomization to minimize the possibility of a bad split. Unfortunately, matching has been shown to decrease the power of the design when the number of pairs is small, unless the matching variable is very highly correlated with the outcome variable (in this case, with change in the health behaviour). We used computer simulation to examine the performance of an approach in which we matched communities but performed an unmatched analysis. If the appropriate matching variables are unknown, and there are fewer than ten pairs, an unmatched design and analysis has the most power. If, however, one prefers a matched design, then for N < 10, power can be increased by performing an unmatched analysis of the matched data. We also discuss a variant of this procedure, in which an unmatched analysis is performed only if the matching 'did not work'. PMID- 7481188 TI - Computing back-calculation estimates of AIDS epidemic. AB - The method of back-calculation estimates the number of HIV infections from AIDS incidence data and projects future AIDS incidence. We explore a conditional likelihood approach for computing estimates of the number of HIV infections and the parameters in the epidemic density. This method is asymptotically equivalent to the usual likelihood method. The asymptotic normal distribution of the estimates facilitates the computation of confidence intervals. We compute standard deviations for the estimates of HIV incidence and project AIDS incidence from the underlying multinomial distributions. We illustrate the methods with applications to AIDS data in the United States. PMID- 7481189 TI - A back-calculation method to estimate the age and period HIV infection intensity, considering the susceptible population. AB - An extension of the back-calculation method to reconstruct the past history of HIV infections is presented. In this method the HIV infection intensity, as a function of age and calendar time, is assumed to belong to a family of parametric functions and is back-calculated from the observed AIDS incidence cases, knowledge of the incubation period and of the susceptible population, the final model being chosen as the one that gives the best fit. The model allows for the interaction between age and calendar time. The application to the main risk groups in Italy, male IVDUs and homosexuals, showed that age and the susceptible population were important in estimating the epidemic among IVDUs; interaction between age and calendar time was found to be significant, showing that among young IVDUs the risk of infection decreased faster than among older IVDUs. Sensitivity of the estimates to the assumption of population and choice of the infection function is investigated. PMID- 7481191 TI - Analysis of ordinal data in a study of endometrial cancer under a matched pairs case-control design. AB - While estimating odds ratios (ORs) in the context of dose levels of conjugated oestrogen exposure and development of endometrial cancer, the categories formed by the levels of the exposure are ordinal in nature. In the literature, the binary logistic model is used for estimating OR for each category relative to the baseline category. We describe the use of two ordinal logistic models, the cumulative logit and continuation-ratio logit models, to estimate the ORs for the matched pairs case-control data set of the Los Angeles endometrial cancer study. A test for equality of the cumulative ORs across the exposure levels is proposed. The test statistic follows asymptotically the chi-square distribution. PMID- 7481190 TI - Estimation of mean sojourn time in breast cancer screening using a Markov chain model of both entry to and exit from the preclinical detectable phase. AB - The sojourn time, time spent in the preclinical detectable phase (PCDP) for chronic diseases, for example, breast cancer, plays an important role in the design and assessment of screening programmes. Traditional methods to estimate it usually assume a uniform incidence rate of preclinical disease from a randomized control group or historical data. In this paper, a two-parameter Markov chain model is proposed and developed to explicitly estimate the preclinical incidence rate (lambda 1) and the rate of transition from preclinical to clinical state (lambda 2, equivalent to the inverse of mean sojourn time) without using control data. A new estimate of sensitivity is proposed, based on the estimated parameters of the Markov process. When this method is applied to the data from the Swedish two-county study of breast cancer screening in the age group 70-74, the estimate of MST is 2.3 with 95 per cent CI ranging from 2.1 to 2.5, which is close to the result based on the traditional method but the 95 per cent CI is narrower using the Markov model. The reason for the greater precision of the latter is the fuller use of all temporal data, since the continuous exact times to events are used in our method instead of grouping them as in the traditional method. Ongoing and future researches should extend this model to include, for example, the tumour size, nodal status and malignancy grade, along with methods of simultaneously estimating sensitivity and the transition rates in the Markov process. PMID- 7481193 TI - Estimation of a common odds ratio under binary cluster sampling. AB - We present results from a simulation study for the estimation of a common odds ratio in multiple 2 x 2 tables when the data are correlated within clusters. We model the correlation of the data by the beta-binomial distribution. Through a simulation study, we compare the Mantel-Haenszel estimator with Rao and Scott's estimator in terms of their biases, observed variances, relative efficiencies of their variances and 95 per cent coverage proportions. We limit the simulation study to the case where there are the same number of subjects in each cluster and the same number of observations in each row of each stratum. When rho = 0, we recommend use of the Mantel-Haenszel estimator gamma MH with an unadjusted variance and Rao and Scott's estimator gamma RSP with a pooled design effect. In general, when rho > 0, we recommend the Mantel-Haenszel estimator gamma MH with an adjusted variance and Rao and Scott's estimator gamma RSP with a pooled design effect. PMID- 7481192 TI - Transient effects in the Cox proportional hazards regression model. AB - We consider a model for mortality rates that includes both the long and short term effects of switching from an initial to a second state, for example, when patients receive an initial treatment and then switch to a second treatment. We include transient effects associated with the switch in the model through the use of time-dependent covariates. One can choose the form of the time-dependent covariate to correspond with a variety of possible transition patterns. We use an exponential decay model to compare the survival experience of transplant versus dialysis treatment of end stage renal disease (ESRD) patients from the Michigan Kidney Registry (MKR). This model involves a hazard function that has an initial effect in mortality at the time of transplant, expected to be higher, followed by a smooth exponential decay to a long term effect, expected to be lower than the risk for those remaining on dialysis. Cox and Oakes used this model to analyse the Stanford Heart Transplant data. The model implicitly suggests there is a time at which the hazard curves (and survival curves) for the treatment groups cross. Those crossing times are useful in advising patients who have the option of receiving a transplant. We describe methods for obtaining estimates of the crossing times and their associated variances, and then apply them in analysing the MKR data. PMID- 7481195 TI - Comparison of tests and sample size formulae for proving therapeutic equivalence based on the difference of binomial probabilities. AB - To prove the hypothesis that a new treatment is as effective as a standard one, a possibility is to test the one-sided null hypothesis of a clear inferiority of the new treatment against the alternative hypothesis that, if at all, it is only negligibly inferior. Such a problem is of clinical relevance if, for instance, a new treatment with an effectiveness which is comparable to that of the standard one would be preferred if it is less toxic. If the difference between the two treatments is measured by the difference of failure rates, approximate statistical tests and sample size formulae have to be used. This paper reports the results of an extensive empirical investigation comparing the well known calculations proposed by Blackwelder, Rodary, ComNougue and Tournade, which propose a sample size formula for the test of Dunnett and Gent, and Farrington and Manning. The investigation was conducted in order to allow more comprehensive conclusions than those which may be drawn from the limited examples given by the last authors. For the usual settings in clinical trials, the formulae of Farrington and Manning are recommended. However, there are combinations of statistical parameters for which they are not preferred. PMID- 7481194 TI - A comparison of size and power calculations for the Wilcoxon statistic for ordered categorical data. PMID- 7481197 TI - Risk ratio and rate ratio estimation in case-cohort designs: hypertension and cardiovascular mortality. PMID- 7481196 TI - An adaptive method for establishing a dose-response relationship. AB - A multistage sampling method is proposed in dose-response trials, where dose adaptions can be performed in the preplanned adaptive interim analyses. The overall test for proving a dose-response relationship is performed by Fisher's product criterion for the p-values from the separate tests of a dose effect in the disjoint samples at the different stages. Based on these p-values decision boundaries for early stopping with the rejection of the global null hypothesis of no existing dose-response relationship are introduced. The power of the adaptive two-stage procedure using a particular adaptation rule is compared with the power of the test for a linear trend under analysis of variance assumptions in extensive simulations. The bias in estimation is also quantified. This procedure could be used for establishing a dose-response relationship without including a placebo treatment. PMID- 7481198 TI - Some perspectives on the career of Samuel W. Greenhouse: the first 75 years. PMID- 7481200 TI - Biostatistical concepts and methods in the legal setting. AB - Biostatistical concepts and methods apply to various problems arising in actual U.S. legal cases. These involve: measures of association, assessing the potential effect of omitted variables and the Peters-Belson approach to regression. In particular, we present the inapplicability of Fisher's exact test in the case where the process determining the marginal sample sizes is not independent of the hypothesis under study by the 2 x 2 table. We adapt Cornfield's procedure to assess whether the omission of another factor could have contributed to a non significant finding of the effect of the major factor under investigation. Finally, we demonstrate the relevance and utility of the Peters-Belson regression methodology to equal pay and promotion cases. PMID- 7481199 TI - Testing independence when the form of the bivariate distribution is unspecified. AB - We address the problem of testing for independence between X and Y in two situations. In the first we assume that the joint distribution of X and Y is unknown but the observations on X and Y are identifiable. In the second case we assume that the distribution of (X, Y) is exchangeable. Here we consider both when (X, Y) are identifiable and when they are not. We illustrate applications to the testing of independence in DNA databases and in twin studies. PMID- 7481201 TI - Biostatistical methodology in clinical trials--a European guideline. PMID- 7481202 TI - Biostatistical methodology in clinical trials in applications for marketing authorizations for medicinal products. CPMP Working Party on Efficacy of Medicinal Products Note for Guidance III/3630/92-EN. PMID- 7481203 TI - The biostatistician in medical research: allocating time and effort. AB - Biostatisticians in research juggle many responsibilities: short-term consulting; long-term collaboration; teaching/training, and statistical research. In an institutionally-supported service group, the biostatistician frequently faces allocation of limited resources (time and effort) over multiple projects, none of which individually supports the biostatistician. In addition to the level of support provided by a specific project, there are several major issues with resource allocation: the quality of the science and data in the project; the possibility that long-term support develops from the work; personal and institutional considerations that involve the specific investigator or project. In this paper, we discuss these considerations along with some examples. We present guidelines for making decisions about allocation of time and effort and the possible implications of setting such priorities. PMID- 7481205 TI - Graphical methods for assessing violations of the proportional hazards assumption in Cox regression. AB - A major assumption of the Cox proportional hazards model is that the effect of a given covariate does not change over time. If this assumption is violated, the simple Cox model is invalid, and more sophisticated analyses are required. This paper describes eight graphical methods for detecting violations of the proportional hazards assumption and demonstrates each on three published datasets with a single binary covariate. I discuss the relative merits of these methods. Smoothed plots of the scaled Schoenfeld residuals are recommended for assessing PH violations because they provide precise usable information about the time dependence of the covariate effects. PMID- 7481204 TI - Effects of model misspecification in the estimation of variance components and intraclass correlation for paired data. AB - Paired data occur in many experimental situations. When one views the subjects as a random sample from some large population, it may seem reasonable to model the data according to the typical one-way random effects analysis of variance (ANOVA). It is then usually of interest to estimate variance components and intraclass correlation. These estimators can be biased if key assumptions are violated, leading to erroneous interpretations and conclusions. We focus upon assumptions about the equality or inequality of means and/or variances of the two measures on each subject. In the framework of the one-way random effects ANOVA model, and three generalizations of it, we document estimators obtained as solutions to the likelihood equations. We consider the potentially serious effects of mistaken assumptions. Our findings suggest that the most general model considered is most desirable if consistent and efficient estimation of the between-subject variance component and intraclass correlation is the main goal. We also briefly connect our exposition to the study of reliability or agreement. PMID- 7481206 TI - Age-adjusted survival curves with application in the Framingham Study. AB - Kaplan-Meier curves provide descriptors of survival information for different subgroups within a data set. Investigators frequently use these figures for descriptive comparison of the effect of a particular measure upon survival. When subjects enter a study at different ages and age is associated with survival, it is often desirable to adjust survival information for age differences in the subgroups. This report discusses several methods for age-adjustment of survival curves, including direct age-adjustment and proportional hazard modelling with age as a covariate. PMID- 7481207 TI - Assessment of cardiovascular risk factors in the elderly: the Framingham Heart Study. AB - In elderly Framingham men and women, systolic blood pressure and cigarette smoking status, as well as a subject's age and sex, strongly influenced the risk of developing cardiovascular disease during ten years of follow-up. Multivariable proportional hazards models were used to assess the roles of several primary risk factors and to examine their secondary effects. The first three factors were noted in both sexes, separately and combined, but the risk function for blood pressure was steeper in men than in women (hazard ratio, HR, 1.53 per 20 mmHg, 95 per cent confidence interval, CI, 1.33 to 1.75 in men; HR = 1.19, 95 per cent CI 1.07 to 1.33 in women). Systolic pressure measured ten years earlier also contributed to CVD risk (HR = 1.16 per 20 mmHg, 95 per cent CI 1.04 to 1.30). even after accounting for current level. Smoking was associated with a 64 per cent elevation in risk, male sex with a 51 per cent increase, and each 5 years increment in age with a 22 per cent increase. Body mass index measured 10 years ago had a modest association, but current body mass index did not. Among diabetic subjects, total serum cholesterol had an asymmetrical U-shaped risk function, the risk increasing to either side of sex-specific median values; diabetes per se, however, was not significant in the final model. In non-diabetic subjects, there was little change in CVD risk up to the median cholesterol values and a modest increase thereafter. None of the risk functions was age dependent. PMID- 7481208 TI - Development of health risk appraisal functions in the presence of multiple indicators: the Framingham Study nursing home institutionalization model. AB - A health risk appraisal function is a mathematical model designed to estimate the risk or probability of a person's mortality or morbidity for various diseases based upon risk factors such as age, medical history and smoking behaviour. The Framingham Study has contributed substantially to the development and use of these for endpoints such as mortality and incidence of coronary heart disease and other cardiovascular diseases. This paper discusses a methodology for the development of health risk appraisal functions when the number of potential risk factors is large and illustrates it with sex specific functions for nursing home institutionalization. The methodology involves grouping variables substantively into sets, applying principal component factor analysis and variable clustering to obtain substantively meaningful composite scores, ranking these in order of substantive importance, and then entering these with a hierarchical ordering into a Cox proportional hazard regression. PMID- 7481210 TI - Assessing goodness-of-fit of parametric regression models for lifetime data graphical methods. AB - Graphical methods are often used to check goodness-of-fit of models to data. It is common to plot residuals against a reference distribution so that when the model fits the data, the configuration should be close to a straight line. Since the resemblance to a straight line is often unclear, it has been suggested to add simulated envelopes within which the configuration is expected to lie. The implementation of this method for survival data analysis is not straightforward. In this paper we point out the difficulties which arise in constructing envelopes on residual plots for randomly censored data. Methods are suggested to deal with the problems and evaluated; they are illustrated by simulated data and on a follow-up study of myeloma. PMID- 7481209 TI - A quality-adjusted survival meta-analysis of adjuvant chemotherapy for premenopausal breast cancer. International Breast Cancer Study Group. AB - The purpose of this paper is to develop and apply a meta-analysis methodology, that does not require patient-level data, for comparing treatments in terms of quality-of-life-adjusted survival. As a motivating example, we considered adjuvant chemotherapy for breast cancer. This therapy has been shown to offer an improvement in recurrence-free and overall survival, especially for younger women, but its acute toxic effects discourage some physicians from prescribing it. To determine whether the benefit of adjuvant chemotherapy treatment outweighs its costs in terms of toxic effects, we performed a meta-analysis of quality adjusted survival based on data from 1229 patients, 49 years of age or younger, randomized in eight clinical trials that compared chemotherapy versus no adjuvant systemic therapy. We conducted the meta-analysis by performing a quality-adjusted survival analysis known as a Q-TWiST analysis on each trial. A Q-TWiST analysis allows one to make treatment comparisons that incorporate differences in quality of life associated with various health states. In this analysis, we define as health states the periods of time patients spend: (i) with subjective toxic effects of chemotherapy; (ii) without symptoms of recurrence and toxicity, and (iii) following disease recurrence. We assigned weights to each health state which reflect their relative value in terms of quality of life and allowed them to vary in a sensitivity analysis. We then combined the individual trial results in a meta-analysis, using a multivariate regression model, in such a way that we could easily perform an overall sensitivity analysis. Individual patient-level data are not required to perform this meta-analysis methodology if the individual Q-TWiST analysis results for each trial are available. PMID- 7481211 TI - Methods for maternal age-standardization of the incidence of congenital abnormalities. AB - In order to compare the birth incidences of particular congenital abnormalities in different populations, it is often necessary to allow for the effects of maternal age. Three age-adjusted indices are defined, based on analogous indices from the literature on mortality studies, namely, the Standardized Mortality Ratio, the Comparative Mortality Figure and Kerridge's Inverse Method. In most practical situations the differences between them are likely to be trival. However, the first index is maximally efficient under multiplicative risk models and is easily adjusted for incomplete data. The second is the only one to provide valid comparisons under additive, as well as multiplicative, models. The third has the advantage that it does not require a knowledge of the maternal age distribution of all births in the population. The use of the three indices is illustrated with published data on Down's syndrome. PMID- 7481212 TI - Predicting the success of in vitro fertilization: conventional semen analysis compared to the hamster ova penetration test. AB - In vitro fertilization has been advocated as a possible treatment for male factor infertility. Using data collected on 154 men, conventional semen analysis and the hamster ova penetration test are compared with respect to their power in predicting the success of assisted conception. As performing the hamster test is more expensive than the semen analysis, we also investigate a combined strategy. It performs better than the discrimination based solely on the semen parameters, but does not lead to an improvement over the overall discriminatory power obtained from the hamster test alone. PMID- 7481213 TI - Analysis of dependent survival data applied to lifetimes of amalgam fillings. AB - When studying lifetimes of amalgam fillings, one is faced with the fact that each patient may contribute multiple survival data. This creates a dependence problem, and some methods for solving it are presented here. Analysis for individual patients with simple approaches for combining data is discussed first. Then a parametric frailty model is applied to analyse variation within and between patients. The results are presented in the form of an empirical Bayes estimate for each patient. PMID- 7481214 TI - On the sample size for one-sided equivalence of sensitivities based upon McNemar's test. AB - In this paper we formalize the problem of testing the one-sided equivalence in the sensitivities of two medical diagnostic tests under a matched-pair study design. We derive conditional and unconditional sample size formulae which are decreasing functions of the probability of being diagnosed by both tests. We calculate upper boundary and midpoint sample sizes. Results of a Monte Carlo simulation study that compares the proposed sample size formulae with that of Lachenbruch suggest that our midpoint conditional sample size is the best choice to obtain the desired power for the type of equivalence studies discussed in the paper. PMID- 7481215 TI - [Clinical examination of the shoulder and diagnostic maneuvers]. AB - The authors review in this article the anamnestic and examination steps of a patient suffering from chronic shoulder pain. They first remind the reader of the muscles intervening in the different shoulder's movements and then go over to the systematic review of the information disclosed by the disorders of the shoulder's motion, whether passively actively or against resistance. They describe the most important diagnosis tests. They finally describe an examination strategy that makes it possible to take a rational clinical decision. PMID- 7481216 TI - [Strategy for the use of imaging in shoulder impingement syndrome]. AB - The authors try to define the relative place of conventional radiology, ultrasound, arthrography, computerized tomography and magnetic resonance imaging in the diagnosis of shoulder impingement syndrome. The above mentioned types of examinations are analysed and commented one by one with their respective advantages and drawbacks. A global approach is described at the end of this article. PMID- 7481218 TI - [Antero-superior impingement of the shoulder. Indications for and options in surgical treatment]. AB - Many therapeutic questions and controversies surround the impingement syndrome of the shoulder, however it appears that the major procedure is rotator cuff decompression. This is achieved by antero-inferior acromioplasty and section of the coracoacromial ligament, during open or arthroscopic surgery. Which patients will be helped by this procedure? When does the surgical decision have to be taken? Is the suture of the rotator cuff tear and the removal of intratendinous calcifications needed in addition to the acromioplasty? Many major questions remain unanswered. PMID- 7481217 TI - [Role of conservative treatment in impingement syndromes of the shoulder]. AB - The impingement syndrome of the shoulder could be primary when the origin is abnormalities of the coracoacromial arch, or secondary when the problem concerns the tendons, the hyperlaxity of the joint or an imbalance between the different muscles. In every case, the treatment has to be first of all conservative, including analgesics, anti inflammatory drugs and rehabilitation. A review of these therapeutic procedures is done. PMID- 7481219 TI - [Hypertrophic cardiomyopathy]. AB - Hypertrophic cardiomyopathy is an uncommon disease, important to recognize. Family screening is mandatory, with physical examination, EKG and echocardiography. Genetic diagnosis will modify our approach in a near future. The different treatments are described: drugs, surgery, pacing. The role of the general practitioner is complementary to the cardiologist's. His task is especially important in the first step of the diagnosis. PMID- 7481220 TI - [Congestive cardiomyopathy]. AB - Idiopathic congestive cardiomyopathy, which is often the result of a silent viral myocarditis, will occur in about 400 patients per year in Belgium. However, a comparable clinical and paraclinical picture, characterized by dilatation of the cardiac chambers and diffuse and profound hypocontractility of the myocardium, can be the common consequence of a large number of cardiac diseases. In front of a clinical presentation of heart failure, the physician will aim to identify the global myocardial insult and to rule out etiologies which, corrected, do not necessarily produce irreversible myocardial damage. This differential diagnosis will be based on medical history, physical examination, echocardiography and coronary angiography. Prognosis of congestive cardiomyopathy, although generally poor, can be greatly improved by a rational pharmacological therapy as well as new interventions (implantable defibrillator, cardiac transplantation) whose correct indications rely on technological advances and a better knowledge of the natural history of the disease. PMID- 7481221 TI - [Myocarditis]. AB - The definition, pathogenesis and clinical picture of myocarditis are reviewed. Recent studies emphasize the importance of immune mechanisms in the genesis of the myocardial damage. Among particular forms of myocarditis, Coxsackie B viral infections, HIV infections, Lyme disease and Chagas disease are successively considered. The main measures for the treatment of myocarditis remain a restriction of physical activities along with the classical medications of heart failure. Due to the increased embolic risk, anticoagulation is often indicated. PMID- 7481222 TI - [Medical and surgical treatment of dilated and viral cardiomyopathies]. AB - Treatment of cardiomyopathy has to take into account correction of etiology or improvement of aggravating factors. Management of cardiomyopathy is not basically different from treatment of classic heart failure. Some drug as amlodipine or miscellaneous beta-blockers might be more effective in cardiomyopathy. Heart transplantation remains the main surgical treatment of end stage cardiomyopathy while other procedures (cardiomyoplasty, ...) are still under assessment. PMID- 7481223 TI - [Electrocardiographic tracings]. PMID- 7481224 TI - [Breast cancer from the general practitioner's viewpoint]. AB - The problem of breast cancer remains a major preoccupation and a challenge for the general practitioner in his daily practice. In a telephonic consultation with several female colleagues, the latter were of the opinion that the master-plan still remains a rapid operation and psychological support after diagnosis. Specialized hospitals are sometimes feared; modern surgery is unappreciated. PMID- 7481225 TI - [Current status and future of diagnosis and therapy of breast cancer: introduction]. AB - The breast cancer in women remains the most frequent with about 30 percent and the rate continues to increase. Happily, parallel to this increase, the diagnosis is done earlier so that the survival is better and treatment becomes less severe. We have several hypotheses for the cause but no one is certain. Risk factors, forecast factors and clinical staging have been established to permit comparable clinical studies and to evaluate treatment efficiency. PMID- 7481226 TI - [Difficult diagnosis in breast cancer]. AB - A detailed senologic evaluation with complete clinical examination, (color Doppler) echography, comparative mammography and cytology or microbiopsy can avoid surgery in case of a post-traumatic fat necrosis, a Mondor Syndrome, an infectious mastitis or a ductal ecstasy with nipple discharge. Actually we have still difficulties to interpret non palpable breast lesions and to supervise operated and radiated breasts. PMID- 7481227 TI - [Fine-needle cytopuncture and/or microbiopsies]. AB - When lesions cannot be clearly identified by sonography or mammography and if their images are clinically occult, their cytology or histology confirmation is required. Stereotactic biopsy allows an accurate locating of non palpable lesions, that are also invisible at sonography. The choice between a fine needle aspiration biopsy and core needle micro-biopsy depends upon local and technical possibilities but above all upon mammographic aspect of the lesion. When microcalcifications have to be identified both methods should be used. PMID- 7481229 TI - [Which surgery for which breast cancer?]. AB - After a brief historical presentation, the two more commonly realized surgical procedure developed in the surgical treatment of breast cancers are described. The relative indications for the mastectomy and the conservative surgery (tumorectomy and quadrantectomy) are explained in different clinical situations. PMID- 7481228 TI - [Surgical approach to subclinical breast lesions]. AB - The treatment of non palpable lesions is one of the most important aspect of breast surgery. Mortality due to breast cancer significantly decreases by about 40% if the tumor is treated before becoming palpable. However, specific modalities have to be strictly respected by a multidisciplinary and experienced medical team. Due to the lack of mass screening organised by public health authorities, physicians in charge of the first line have to generously prescribe mammograms to their patients aged 50-70. Expertise is required for the second line. An expert radiologist is a radiologist who is able to detect subclinical breast lesions and to precisely locate them within the breast. An expert surgeon is an experienced one who can read a mammogram, locate a non palpable lesion and completely remove it according to radiologist information. The pathologist refusing a frozen section and having a look on the mammogram to understand the medical story is the good one, allowing an appropriate therapy based on a precise and complete histologic diagnosis. PMID- 7481230 TI - [Role of axillary assessment and current technical aspects in the surgical treatment of breast cancer]. AB - In case of invasive breast cancer, the pathologic assessment of axillary lymph nodes status gives information on staging, prognosis and on the use of systemic adjuvant therapy. Removing at least ten axillary lymph nodes seems to prevent complications arising from uncontrolled tumor growth in the axilla. An axillary dissection is not justified in ductal carcinoma in situ (DCIS). To assess new techniques of axillary dissection will probably help to decrease the complications of the classical approach with oncological safety. Liposuction and endoscopic biopsies seem to give good results in preliminary studies. PMID- 7481231 TI - [Breast cancer: apropos of "in situ"]. AB - Since the introduction of screening mammography, the proportion of ductal carcinoma in situ (DCIS) has significantly increased. Their early detection result in the majority of the cases in an exclusive radiologic expression and a histopathologic measure inferior to 20 mm. Newly formulated proposals of histologic classifications may permit in the future to identify subtypes of DCIS with a different clinical behaviour and prognosis. In analogy to the treatment's evolution for invasive mammary cancers, the concept of conservative breast surgery is evaluated in prospective randomized trials, together with the impact of adjuvant radiotherapy. Preliminary results report high survival rates, despite a considerable number of local recurrences. Radiotherapy seems not to reduce proportionally the risk for non-invasive and invasive recurrences. PMID- 7481232 TI - [Breast reconstruction following mastectomy]. AB - In our country reconstructive breast surgery developed approximatively 20 years ago. In the early 70s breast reconstruction was only performed by simple implantation of a prosthesis. Other surgical techniques were quickly introduced, such as the transfer of the musculocutaneous flap of the latissimus dorsi and the autogenous reconstruction with the rectus abdominis musculocutaneous island flap, better known as TRAM. Since the beginning, breast reconstruction was recommended after a minimal delay of one year. More recently reconstruction was proposed to start earlier and immediate breast reconstruction became indicated in certain specific instances. PMID- 7481233 TI - [What is the role of radiotherapy in the treatment of breast cancer in 1995?]. AB - In 1995, radiotherapy continues to remain one important component in the treatment of breast cancer both for the breast conserving approach and after a mastectomy in case of more locally advanced disease: it helps to improve the local control and so to safe more breast or to avoid a relapse in the mastectomy scar, a source of distress for the woman. Nevertheless, this modality requires a precise technique and integration with surgery and chemotherapy. Besides this curative approach, radiotherapy plays an important role in the treatment of brain or bone metastases where it is quite useful to improve the patient's quality of life. PMID- 7481234 TI - [Respective indications for hormone therapy, induction chemotherapy and adjuvant chemotherapy in breast cancer without clinically discernable distant metastases]. AB - Adjuvant therapy for breast cancer is an effective mean to reduce the likelihood of recurrence and to increase overall survival, as shown by the Early Breast Cancer Trialists' Group Overview. Although these benefits are clearly significant, they represent a modest improvement in the care of breast cancer. The role of pre-operative chemotherapy remains to be defined in early stage breast cancer. Otherwise, pre-operative chemotherapy has become 'standard of care' in locally advanced and inflammatory breast cancer. There are still several questions to be answered in the field of adjuvant and pre-operative chemotherapy. Prospective randomized clinical studies are the only way to obtain further therapeutic progress for this highly lethal disease. PMID- 7481235 TI - [Psychological aspects of breast cancer]. AB - The announcement of the diagnosis of breast cancer is always distressing. With breast cancer, the symbol of femininity is compromised. The loss of the breast, partial or total, will interfere with the identity of the woman. The psychological impact of breast cancer varies from a woman to another. It depends of the age, the patient's personality, the real life experience, her social and familial relationships, her psychological or psychiatric history. But the way the disease will be diagnosed and the given treatments may also influence the pattern of the psychological impact. Every time a cancer is diagnosed and especially a breast cancer for a woman, the psychological history should be taken into account. In our discussion, we will talk about the psychic scene invaded by a sick body. And like every story, it starts with "Once upon a time ...". PMID- 7481236 TI - [Neurological complications of diabetes]. AB - One of the hallmarks of diabetes mellitus is its propensity to cause neurological complications. Diabetes is an independent risk factor of stroke. Diabetic neuropathy represents the most common type of peripheral neuropathy in our country. Improved glucose control can improve nerve function and restoration of the euglycemic state appears to stop the progression of the neuropathy. Treatment strategy of painful neuropathy with tricyclic antidepressant, anticonvulsant, anesthesia agents and topical capsaicin is discussed. PMID- 7481237 TI - [Diabetic retinopathy]. AB - Diabetic retinopathy remains the most frequent cause of blindness in the active population of industrialized countries. However, decreased vision and blindness can efficiently be reduced by good metabolic control, yearly retinal examination since onset of diabetes from puberty onwards, and timely laser photocoagulation. Increased frequency of retinal controls, in case of retinal or general risk factors, enables to detect and to treat threatening lesions early, leading to reduction of decreased vision and blindness. PMID- 7481238 TI - [Renal complications of diabetes]. AB - Many diabetic patients will develop a nephropathy that will eventually result in end-stage renal failure. The early stage ("incipient") of diabetic nephropathy generally appears after 5 to 20 years of diabetes and is characterized by microalbuminuria (30 to 300 mg/day), which is only detectable by sensitive radio immuno-enzymatic methods. When a frank proteinuria develops (> 500 mg/day), the glomerular filtration rate inexorably declines, resulting in terminal renal failure after several years. The onset of microalbuminuria or the elevation of blood pressure (above 120-140/80 mmHg) are predictive of a poor evolution and require appropriate preventive therapeutic interventions. These include an optimal control of hyperglycaemia, dietary proteins and salt restriction, and prescription of anti-hypertensive drugs, with a particular benefit ascribed to angio-tension converting enzyme inhibitors (and maybe to certain calcium channel blockers). These interventions have been proven efficient to prevent or slow down the evolution of diabetic nephropathy. PMID- 7481239 TI - [Vascular complications of diabetes]. AB - Vascular complications are common in diabetics. Patients with type I diabetes mellitus may be more likely to suffer microvascular complications (retinopathy, nephropathy, neuropathy), whereas patients with type II diabetes may be more likely to have macrovascular complications (similar to atherosclerotic disease). Amputation rates are 10 to 15 times more common in the diabetic than in the non diabetic. A meticulous examination (historical and physical) combined with doppler ultrasonography will indicate the presence of peripheral vascular disease in the diabetic. Such an evaluation enables correct management of the risk factors and ensures an appropriate follow-up and treatment of the diabetes related lesions as soon as they occur. PMID- 7481240 TI - [Daily aspects of proctology]. AB - Revue of the commonest anal diseases encountered in the daily practice of general medicine. Hemorrhoids share their symptoms with a whole series of other diseases, and it is this lack of specificity that always calls for a thorough examination to reach a precise diagnosis. The most usual and accessible methods of treatment are described. Anal fissures: their diagnosis and, mostly, their prompt conservative treatment lead to frequent therapeutic success. Anal fistulas: if their description and origins are interesting, it is mostly the prompt incision and drainage of the abscess, by the general practitioner, without use of antibiotics, that will prevent the formation of fistulas. Pilonidal sinuses must be recognized as incurable without the help of surgery. Condylomas can often be treated at the office. PMID- 7481241 TI - [Anorectal tumor pathology and its diagnosis]. AB - Anal cancer is very rare, but its diagnosis is often too late. Treatment is usually non surgical and can be very effective. Prognosis can be improved by earlier diagnosis. Proctoscopy is mandatory for all symptomatic patients. Could an high risk population be determined, the incidence of anal cancer could fall. By contrast, rectal cancer is one of the most frequent and its prognosis has not been improved. Hope can be found in screening and prevention; these will be more effective when the transformation sequence of adenoma to cancer will be entirely explained. PMID- 7481242 TI - [Anorectal localization of chronic inflammatory diseases of the intestines]. AB - In Ulcerative Colitis (UC), the rectum is always involved, but the disease may be extended to the left colon (distal colitis) or the entire organ (pancolitis). Secondary perineal lesions (erythema, fissures, hemorrhoids) are limited to local irritation related to the diarrheic syndrome; specific ano-rectal lesions are related to the mucosal inflammatory process, but others pathological situations may be observed and are due to the postoperative status (stenotic ileo-anal anastomosis, pouch inflammation). Ano-rectal localisation of Crohn's Disease (CD), sometimes underestimated, is clinically more complex (abscess, fistula, ulcer, etc) and requires precise classical (endoscopy, conventional radiology) or more modern (echo-endoscopy, nuclear magnetic resonance) investigations. The treatment is more difficult (medical conservative, "a minima" or more extended surgery). Differential diagnosis, symptoms, methods of investigation and appropriated therapeutical choices are successively developed. PMID- 7481243 TI - [Bone following the menopause]. AB - Bone is a tissue undergoing permanent remodeling, submitted to the influence of endocrine and local (or paracrine) factors. After reaching a peak, bone mass decline physiologically with age, showing a more important decrease during early postmenopausal years. Bone remodeling tightly couples bone resorption due to osteoclasts to bone formation due to osteoblasts. Both osteoclasts and osteoblasts originate from bone marrow and therefore synthesize and respond to common cytokines such as interleukin 1, 6, 11 or to growth factors as colony stimulating factors (CSF). The understanding of the mechanisms that regulate osteoclasts and osteoblasts activation both in physiological situation or during the loss of gonadal functions is in progress. In the next future, it would be possible to give a more efficient and rational therapy than today. PMID- 7481244 TI - [Cardiovascular effects of replacement hormones]. AB - The authors discuss the possibility of a consensus concerning hormone replacement therapy in relation with the risk of cardiovascular diseases in postmenopausal women anno 1995. Although there are not yet randomised clinical trials published concerning the effect of hormone replacement therapy, combined or not, on the incidence of coronary heart disease, the criteria for causal association seem to be indeed present. Relative risk of women on replacement therapy compared to those without such therapy is around 0.55; there is a gradient which expresses itself by an inverse correlation between the length of the hormonal replacement therapy and risk of coronary heart disease; this substitution therapy is most probably an independent protective factor; there are numerous possible pathogenetic mechanisms, the most important being likely a significant increase of HDL-cholesterol; animal experiments confirm the favourable effects of natural oestrogens on atherogenesis. Finally, a recent clinical trial seems to show that the favourable effects on the coronary risk profile are observed both with and without combined hormonal substitution therapy. PMID- 7481245 TI - [Estrogens following menopause. Methods of hormone replacement]. AB - Even today hormone replacement therapy concerns still only a rather small number of post-menopausal women: 19% of women aged 50-75 on the basis of prescriptions. The compliance of oestrogen substitution reflects even more a general reticence against the use of hormones within the public opinion and among physicians. Thus, hard work for widespread information remains to be achieved, based on scientifically well established data. The purpose of this presentation is to describe benefits and drawbacks of the modes of administration of oestrogens and of oestro-progestative associations and to consider the new developments in this matter. PMID- 7481246 TI - [Compliance with hormone replacement therapy]. AB - Hormone Replacement Therapy (HRT) after menopause reduces climacteric symptoms and increases the quality of life. If correct dosage is used, bone loss can be prevented and fracture frequency reduced. Epidemiological studies also suggest a decreased cardio-vascular mortality among HRT users. A high proportion of post menopausal women should therefore be treated using long-term therapy, but currently only a small proportion of women use HRT in Belgium, (around 14%). Further their compliance is particularly low. Fear of cancer and drug-related side effects, such as unaccepted bleeding are often mentioned causes of low compliance. Education of both physicians and patients, regarding HRT seems to be a crucial issue. The physician's attitude towards HRT is often reported as negative. Osteoporosis prevention and related bone mass measurements may have a favorable effect on compliance, but this is not well established. Establishing a confidential relationship with patients and the involvement of nursing counseling are strategies which may improve compliance. The use of easy-to-take medications supported by calendar blisters or reminders, may facilitate chronic use of medication. Future research needs to investigate the physician's and the patient's decision-making process. PMID- 7481247 TI - [Methotrexate treatment in non-cancerous disorders]. AB - Weekly administration of low doses methotrexate (7.5 mg to 25 mg a week) is considered as one of the most important progresses in the management of rheumatoid arthritis. Its efficacy has also been demonstrated for the treatment of other inflammatory processes such as psoriasis arthritis and, more recently, Crohn's disease. We here review the indications, the limits and the side effects of such therapeutic approach in various inflammatory and/or autoimmune diseases. PMID- 7481248 TI - [The treatment of deep venous thrombosis with low-molecular-weight heparin]. AB - The authors, via a long review of published studies, show that low-molecular weight heparin, with early start of antivitamin K, is actually the treatment of choice in thromboembolic disease. PMID- 7481249 TI - [Leading drugs in 1995: success, failures and uncertainties]. AB - Presentation of drugs most worthy of interest in autumn 1995. Among these, the ACE inhibitors, the HMGCOA inhibitors, the Proton Pump inhibitors, the serotoninergics used against depression and migraine, the endobronchial corticoids and finally the ASA. Review of their successes, failures and uncertainties. PMID- 7481250 TI - [Health economics: viewpoint of a general practitioner]. AB - Health economics is a quite old discipline, linked to Public Health. It suffers a prejudice because of the circumstances in which it was reintroduced in the programs of continued medical education. The physician should be informed about and should deal with the development of this discipline. PMID- 7481251 TI - [Health economics and antibiotic therapy]. AB - In the field of antibiotic therapy, particularly the methods of economic evaluation hold one's attention within the wide range of health economics' applications. Several tools allow a comparison of the outcomes of alternative strategies and thereby guide choices to the most appropriate solutions. After a brief recall of the methods classically used to evaluate health care strategy, the authors stress the importance and difficulty of fixing and applying a correct and satisfactory procedure for evaluation. An evaluation example of antibiotic therapy allows to illustrate the application of the principles confronting a field in which competition is intense and economic stakes stay large--a fact which naturally yields to seek after objective decision making criteria. The health care policies drawn by public authorities as well as the marketing strategies of the health sector trade are partly based on such evaluations. If these techniques are not intended for the practitioner in the first place, they should not be indifferent to him since they influence health authorities and thereby indirectly affect the therapeutic freedom of the physician. PMID- 7481252 TI - [Economic approach to breast cancer screening in Belgium]. AB - In Belgium, the breast cancer screening by mammography involves a great number of women. It is largely non organized and regarded to the small number of data, it is impossible to determine if this screening is useful and if it has a good "cost benefit" ratio. This work studies the economical aspects of breast cancer screening in the belgian health care system. From information found in other countries, we build 4 models corresponding to an organized and a spontaneous screening. We studied the total processus from screening to diagnosis, including the quality assurance and the evaluation of effectiveness in the organized models. We then applied the reimbursements of the belgian health insurance in the models and compared the costs. It appears that a screening for breast cancer must be organized to give a best "cost-effectiveness" ratio. Pilot projects should be the best way to study the best organization modalities in Belgium. PMID- 7481253 TI - [Practical attitude in the matter of screening in the general practitioner's office]. PMID- 7481254 TI - [The right choice in medical imaging studies. General practitioner's viewpoint]. AB - Different criteria are shown, able to guide the general practitioner in his choice of schemes of investigations for diagnosis in radiology respecting the realities of the patient and of the socioeconomic surrounding. PMID- 7481256 TI - [Pharma-clinics. How I treat... a patient with atrial fibrillation. 2]. PMID- 7481255 TI - [The "right choice" in medical imaging studies: the radiologist's viewpoint- application to lumbosciatic pain]. AB - These past few years, very efficient imaging technologies, which are not traumatic for the patient, but often quite expensive, have been developed. Within the context of budgetary restrictions, these progress require a carefully planned strategy on the choice of investigations. First of all basic rules are defined: "the most simple act", "the direct way", "the direct exploitation of results", "the therapeutic relevance", "the multidisciplinary integration". Several erroneous types of behaviour are also described: "the most comprehensive set of examinations", "the examination in fashion", "the exaggerated medico-legal fear". Finally, these principles apply to a common clinical situation: lumbosciatic pain. Relevant examinations are first spotted: standard examination, myelography, discography, CT scan, MRI. A synthetic diagnostic approach is then suggested. The latter rules out any complementary study in the case of non-chronic common lombosciatic pain and in the absence of certain revealing signs. In the other cases, the correct combination of conventional radiology, nuclear medicine, CT and MRI is suggested, according to the clinical context. PMID- 7481257 TI - [Clinical case of the month. Growing cutaneous nodules in a 60-year-old]. PMID- 7481258 TI - [Current therapeutics 1990-1995 in the cardiovascular field]. PMID- 7481259 TI - [Arterial hypertension in children]. PMID- 7481260 TI - [Cervical herniated disk]. PMID- 7481261 TI - [Deep venous thrombosis of the upper limbs. Literature review. Apropos of a case of Paget-Schrotter syndrome]. PMID- 7481262 TI - [Internuclear ophthalmoplegia and sickle cell trait: apropos of a case]. PMID- 7481263 TI - [How I examine... global dilatation of the cerebral ventricles in an elderly patient]. PMID- 7481264 TI - [Pharma-clinics. Drug of the month. Terbinafine (Lamisil)]. PMID- 7481265 TI - [Pharma-clinics. How I treat... an erectile deficiency]. PMID- 7481267 TI - [Arterial hypertension and vascular resistance]. PMID- 7481266 TI - [Clinical case of the month. Prolonged spontaneous remission of a typical type 1 diabetes]. PMID- 7481268 TI - [Iatrogenic encephalopathies due to non-psychotropic drugs]. PMID- 7481269 TI - [Digestive system tuberculosis in the Liege area]. PMID- 7481270 TI - [Clinical significance of bronchial hyperreactivity]. PMID- 7481271 TI - [Surgical gloves, condoms, rubber balloons and other latex objects. A look at their pathogenic aspects]. PMID- 7481272 TI - [Cognitive distortion in the affirmation of self. Various observations]. PMID- 7481273 TI - [How I explore... fever of unknown origin]. PMID- 7481274 TI - [Pharma-clinics. Drug of the month. Azithromycin (Zitromax)]. PMID- 7481275 TI - Physiological monitoring of elite cyclists. Practical methods. PMID- 7481276 TI - Vitamin E status and response to exercise training. AB - Vitamin E is an important intramembrane antioxidant and membrane stabiliser. Over the past 40 years, vitamin E supplementation has been advocated for athletes in the hope of improving performance, minimising exercise-induced muscle damage and maximising recovery. However, there is currently a lack of conclusive evidence that exercise performance or recovery would benefit in any significant way from dietary vitamin E supplementation. Exceeding current recommended intakes of vitamin E even by several orders of magnitude will result in relatively modest increases in tissue or serum vitamin E concentrations. Most evidence suggests that there is no discernible effect of vitamin E supplementation on performance, training effect or rate of postexercise recovery in either recreational or elite athletes. There is very little evidence, particularly involving humans, that exercise or training will significantly alter tissue or serum vitamin E levels. While there is some evidence that certain indices of tissue peroxidation may be reduced following dietary vitamin E supplementation, the physiological and performance consequences in humans of these relatively minor effects are unknown. Although there appears to be little reason for vitamin E supplementation among athletes, it does not appear that the practice of supplementation is harmful. PMID- 7481278 TI - Aging, physical activity and sports injuries. An overview of common sports injuries in the elderly. AB - Illness and aging both cause many structural and functional alterations in the human body, rendering elderly people liable to overloading of the musculoskeletal and cardiovascular systems. It should, however, be kept in mind that immobilisation and inactivity have even more deleterious effects on structures and functions in the elderly than in younger adults. Most physically active elderly people are selected individuals with respect to their superior health and physical capacity compared with inactive persons of the same age, thus making it possible to further improve their physical capacity. They will, however, be affected by some of the drawbacks of physical overloading, mostly due to the diminished ability of aging body systems to adapt to high levels of loading. The safety margin of an exercise dose tends to decline with aging. Exertional injuries are common among the elderly, and are connected mostly with degenerative aging processes. Acute injuries are common in those elderly people participating in sport activities which demand high coordination, reaction time, and balance capabilities, such as ball games, down-hill skiing, and gymnastics. Muscle has been reported to be the most commonly acutely injured tissue among active elderly athletes. The lower extremities are the most susceptible to injury. A large proportion of injuries (acute and exertional) are mild and can be treated by brief cessation of training and competition activities. Some of the injuries are, however, long term and cause disability not only during training and competition, but also in the normal activities of daily living. It is important that these injuries are treated as soon as possible and in the most effective way, similarly to injuries suffered by younger people. In treating elderly people, it is most important to avoid the detrimental effects of immobilisation; this requires active treatment and rehabilitation with compensatory exercise therapy. The best 'treatment' for sports-related injuries is prevention. Good agility, technical skills, and cardiovascular and musculoskeletal fitness are important in injury prevention among the elderly. Appropriate training programmes, the use of safe and familiar equipment, careful warming up and cooling down, multiphasic training [including the training of neurophysiological functions (balance, coordination and reaction time)] and muscle strength are essential aspects of injury prevention. PMID- 7481277 TI - Delayed muscle soreness. The inflammatory response to muscle injury and its clinical implications. AB - Delayed onset muscle soreness (DOMS) is a sensation of discomfort that occurs 1 to 2 days after exercise. The soreness has been reported to be most evident at the muscle/tendon junction initially, and then spreading throughout the muscle. The muscle activity which causes the most soreness and injury to the muscle is eccentric activity. The injury to the muscle has been well described but the mechanism underlying the injury is not fully understood. Some recent studies have focused on the role of the cytoskeleton and its contribution to the sarcomere injury. Although little has been confirmed regarding the mechanisms involved in the production of delayed muscle soreness, it has been suggested that the soreness may occur as a result of mechanical factors or it may be biochemical in nature. To date, there appears to be no relationship between the development of soreness and the loss of muscle strength, in that the timing of the two events is different. Loss of muscle force has been observed immediately after the exercise. However, by collecting data at more frequent intervals a second loss of force has been reported in mice 1 to 3 days post-exercise. Future studies with humans may find this second loss of force to be related to DOMS. The role of inflammation during exercise-induced muscle injury has not been clearly defined. It is possible that the inflammatory response may be responsible for initiating, amplifying, and/or resolving skeletal muscle injury. Evidence from the literature of the involvement of cytokines, complement, neutrophils, monocytes and macrophages in the acute phase response are presented in this review. Clinically, DOMS is a common but self-limiting condition that usually requires no treatment. Most exercise enthusiasts are familiar with its symptoms. However, where a muscle has been immobilised or debilitated, it is not known how that muscle will respond to exercise, especially eccentric activity. PMID- 7481279 TI - Effectiveness of prophylactic ankle stabilisers for prevention of ankle injuries. AB - Ankle injuries are common at many levels of athletic participation. A relatively recent approach in injury intervention is the use of prophylactic ankle stabilisers (PAS). PAS are used with the intention of reducing the frequency and severity of ankle injuries in a cost-effective manner. To date, 4 studies have been completed to determine the clinical efficacy of PAS. Although all of the studies have methodological limitations, a general consensus of agreement exists among the findings: PAS are effective in reducing the incidence of acute ankle sprains. However, the effect of PAS on ankle sprain severity remains unclear, as varying results have been reported. PAS do not increase the risk of knee injuries. The use of PAS for ankle injury reduction appears to be justified although further research is required. PMID- 7481280 TI - Gastro-oesophageal reflux and exercise. Important pathology to consider in the athletic population. AB - Gastro-oesophageal reflux is commonly found in the general population, and has recently been demonstrated to occur more frequently during exercise than at rest. This fact is significant to the substantial number of athletes who complain of exertional upper gastrointestinal symptoms and exercise-induced chest pain. A diagnosis of exercise-induced gastro-oesophageal reflux can be confirmed by means of ambulatory pH monitoring. A positive diagnosis allows for appropriate management of the individual. This can involve simple measures, such as recommendations for changes in diet, timing of meals, and nature of exercise. However, pharmacological intervention may be required. A decrease in morbidity associated with cardiac origins of exercise-induced pain can also be expected with a more comprehensive understanding of this pathology. PMID- 7481281 TI - Anterior cruciate ligament injuries in young athletes. Recommendations for treatment and rehabilitation. AB - Complete midsubstance tears of the anterior cruciate ligament (ACL) in skeletally immature adolescents are being diagnosed and reported with increasing frequency. Conservative treatment of such ACL tears in junior high schools, subsequent meniscal tears, and very early osteoarthritis. Results of primary repair of the torn ACL and extra-articular reconstructive procedures have been disappointing. Intra-articular ACL reconstruction in young athletes approaching skeletal maturity using the autogenous patellar tendon graft gives excellent knee stability, and decreases the risk of meniscal tears. Athletes are able to return to competitive sports at their preinjury level. Here, the existing literature on the subject of midsubstance ACL tears in young athletes is reviewed, and guidelines for the management of ACL-deficient knee in this population are provided. PMID- 7481282 TI - Growth and development of young athletes. Should competition levels be age related? PMID- 7481283 TI - Gastrointestinal symptoms during exercise. The effect of fluid supplementation. PMID- 7481284 TI - Exercise loading of tendons and the development of overuse injuries. A review of current literature. AB - This review examines recent studies on the effects of exercise on tendons in animal models. Although tendon adaptation to exercise has been described using histology, morphometry, ultrasonography and molecular biology, precise measurements of excess tendon loading during exercise protocols have not been reported. Only a few studies have attempted to evaluate the mechanical strength of exercised tendons. The long term effect of exercise on tendons appears to be positive, but researchers have suggested that periods of mechanical weakness occur in tendons during adaptation to loading conditions. Studies documenting changes associated with the terminal state of pathological tendons are also summarised. Unfortunately, there are no descriptions of tendon tissue in the early stages of overuse injury. Since blood flow is commonly implicated in the emergence of tendinitis, the final section covers recent work on blood flow and tendon physiology. Related research identifying cellular mediators (hyperthermia, hypoxia, and oxidative stress) involved in the development of tendinitis is also presented. Suggestions for further research into exercise loading and the development of tendon overuse injuries are made. PMID- 7481287 TI - [Patients with lung diseases and sports]. AB - Lung patients are suffering from dyspnea. These symptoms make any physical exertion unpleasant, and patients become sedentary. This behaviour leads to a deconditioning of the cardiocirculatory and muscular system which renders activity even more unpleasant, thus reinforcing the sedentary life style. This downward spiral can be interrupted by a program of progressive exercise which should be adapted to the type and severity of the pulmonary disease. In this context, the value of sport therapy in the treatment of asthma bronchiale and COPD is redefined. The possibilities and limitations for this form of treatment are explained. PMID- 7481286 TI - [What is your diagnosis? Situs inversus with incomplete right bundle-branch block]. PMID- 7481288 TI - [Sports and nutrition]. AB - A varied, well-balanced, healthy diet, rich in carbohydrates, is one of the cornerstones of sports nutrition. In addition, an equilibrated supply in energy and nitrogen, the ideal distribution of the nutrients (6 to 9 g carbohydrates per kilogram body weight, 1 to 1.5 g fat per kilogram body weight, 1.2 to 1.8 g protein per kilogram body weight), a well-balanced fluid intake and the choice of food with a high density of nutrients constitute an important cornerstone. All the forementioned elements, which are contained in a basic nutrition, guarantee the needs of the micro- and macronutrients in combination with a cautious substitution of minerals, trace elements and vitamins. The endurance capacity is supported by this high-quality basic diet which can be modified minimally during performance and for regeneration. PMID- 7481289 TI - [Doping]. AB - The use of performance-enhancing drugs and methods has been known for centuries. Man always tried to expand his given natural limits. In sport, these practices are called 'doping' and are forbidden by the rules. Today, doping is the advertent or inadvertent use of substances of forbidden classes or of forbidden methods defined by the actual doping list. This doping list is a so-called 'open' list that does not mention all substances within a forbidden class, but only some examples. This imposes high demands on physicians treating athletes. The Commission against Doping of the Swiss Sports Federation helps in case of questions about medical treatments. The Swiss Federation of Sports Medicine confirmed lately its fight against doping with a formal position paper. PMID- 7481285 TI - Methodological and statistical considerations for exercise-related hormone evaluations. AB - Improvements in laboratory techniques have allowed research related to exercise endocrinology to flourish. The emerging literature, however, is often inconsistent and contradictory. The discrepancies in research findings are possibly the result of poor control of confounding variables and/or inappropriate methodologies or analyses. Environmental and pretesting behavioural conditions must be standardised to minimise the influence of variables not directly related to the investigation. Environmental temperature and relative humidity, alcohol, caffeine and nicotine intake, prandial state, sleep deprivation and previous exercise can each alter hormonal responses to exercise. Both prescription and over-the-counter medications can also modify normal hormonal secretions thereby confusing exercise-induced findings. Specimen collection and analysis procedures must be controlled carefully. Changes in plasma volume related to postural changes or tourniquet-induced stasis can confound attempts to isolate exercise related endocrine responses. The established circadian and rhythmical variations characteristic of many hormones need to be controlled. The specimen selection (plasma, serum, urine, etc), collection, storage and analysis procedures should be carefully planned and evaluated. The magnitude of haemolysis, analytical and biological variation must also be monitored. Isolating the hormonal perturbations resulting from a particular exercise variable can be very difficult. Exercise intensity, duration, mode, frequency and volume may each have specific effects on the endocrine changes seen with exercise and training. Furthermore, hormonal responses to exercise are dependent upon initial training status and fitness level. The statistical procedures and data presentation options selected to convey experimental findings can bias experimental results. The descriptive and inferential statistics to be used for data analysis should be preplanned and consistent with the underlying assumptions of the analytical procedure. Careful consideration should be given to the biological relevance of statistically significant findings. In some cases, data transformations (e.g. absolute vs relative changes, logarithmic) should be considered for analysis or presentation. Given the individual nature of hormonal responses to exercise, emphasis should be placed presenting individual data. Other considerations, including age, sex, racial origin and disease conditions need to be controlled for when trying to examine exercise-induced hormone changes. PMID- 7481290 TI - [A case from practice (331). Bilateral sensory carpal tunnel syndrome more pronounced on the right side]. PMID- 7481291 TI - [Hypoglycemia: diagnostic approach]. AB - Hypoglycaemia can occur if the endogenous liver glucose output is lower than the glucose uptake in insulin-sensitive and insulin-insensitive tissues. The onset of hypoglycaemia induces the production of counterregulatory hormones such as glucagon, epinephrine, growth hormone and cortisol, and symptoms of neuroglycopenia. The correlation between biological hypoglycaemia and the symptoms associated with low blood sugar is particularly poor in diabetic patients and in patients with suspected postprandial hypoglycaemia. It is important to discriminate between fasting and postprandial hypoglycaemia. Idiopathic postprandial hypoglycaemia should be diagnosed clinically without further laboratory assessment, whereas the etiology of a fasting hypoglycaemia needs to be clarified further by laboratory testing, as it is potentially life threatening. PMID- 7481292 TI - [Interdisciplinary therapy of Ewing sarcoma]. AB - Ewing's sarcoma is the most common bone tumor in children under ten years of age. It represents a highly malignant tumor that usually arises in bone, but occasionally occurs in soft tissues. Most patients with Ewing's sarcoma present with what appears to be localized disease, but surgery and radiotherapy alone is unlikely to be curative due to the presence of micrometastases. Consequently chemotherapy has become increasingly effective in eliminating these cells, as well as assisting in obtaining local control. The goal of the optimal integration of the three modalities of therapy is to eradicate all tumor while preserving as much function as possible. The extent of the disease determines the aggressiveness of treatment. Diagnostic evaluation includes modern radiologic imaging, immunochemical and cytogenetic techniques. The outlook has dramatically changed in case of nonmetastatic disease, achieving local control of the tumor in 80%, and producing 5-year disease-free survival in two thirds of the patients. PMID- 7481293 TI - [Cyst of the major omentum--laparoscopic diagnosis]. AB - This report describes the case of a 22-year-old female patient with noncharacteristic complaints in the lower abdomen. Sonography revealed a large cyst close to the right adnexa, suggestive of adnexitis; however, laparoscopy, performed because of suspected adnexitis, revealed a cystic tumor connected to the omentum instead. PMID- 7481295 TI - [What is your diagnosis? Right ventricular hypertrophy]. PMID- 7481294 TI - [A case from practice (332). Giardia lamblia infection of the gastrointestinal tract]. PMID- 7481296 TI - [Clinical video-capillary microscopy]. PMID- 7481298 TI - [Initiation of treatment in Parkinson disease]. AB - Initiating treatment in a patient with Parkinson's disease requires consideration of age, degree of disease activity, and consequences of long-term treatment. In young patients, albeit dopaminergic agonists or selegiline are possible alternatives, they will never be as effective as L-dopa, and they can be used only initially in very mild cases and for a short period of time. Long-acting L dopa is probably better after titration with standard preparations. Potential neuroprotection by selegiline has not been confirmed so far, but it acts as a mild anti-Parkinsonian. In patients over 60, it is recommended not to use anticholinergic or dopaminergic agonists, but to start with a low dose of L-dopa and increase it by a maximum of 125 mg every 7 to 10 days. Clozapine can be very useful against psychosis. PMID- 7481299 TI - [Therapeutic immunomodulation in neurology: from hope to reality. Exemplified by multiple sclerosis]. AB - Multiple strategies for specific therapies are presently attempted in multiple sclerosis, in which an autoimmune reaction leads to demyelinisation within the central nervous system. Similar approaches are generally used in other potential autoimmune diseases. They try to interfere with the presentation and recognition of putative autoantigens by the immune system and also with the inflammatory process that gives rise to the demyelinating lesion. The authors review extensively these various prospects and their immediate implications for multiple sclerosis patients. PMID- 7481297 TI - [Reiter's disease--a review of an interdisciplinary disease]. AB - Over the past 20 years the term 'Reiter's disease' has been expanded from the classical triad of sterile urethritis, conjunctivitis and arthritis to include a broader range of reactive arthritides. Indeed, the classical syndrome as described by Reiter is rarely seen in its complete form, and the presence or absence of active infection in the joint is being redefined in the light of recent techniques of molecular biology. The terminology of Reiter's syndrome, reactive arthritis and seronegative spondylarthropathy are defined here using case examples. PMID- 7481300 TI - [Thrombolytic therapy in ischemic cerebrovascular accidents]. AB - Thrombolysis together with neuroprotective agents the two most important recent advances in the treatment of acute ischemic stroke. This update summarizes the recent randomized clinical trials and emphasizes the question of risk versus benefit of this potentially useful but dangerous treatment. PMID- 7481302 TI - [Current viewpoints in the treatment of epilepsies: role of new antiepileptics?]. AB - Although standard anticonvulsants are effective in achieving complete seizure control in the majority of patients, an appreciable proportion (about 20 to 25%) is at least in part resistant to conventional pharmacotherapy. Efficacy of carbamazepine, phenytoin, phenobarbital and valproate is very similar. Should one drug fail because of inadequate efficacy and unacceptable adverse effects, an alternative monotherapy should be used. Surgical treatment is a possible therapeutic option for only some of these patients. The development of newer, more effective drugs, such as vigabatrin, lamotrigine, gabapentin and oxcarbazepine, for monotherapy is desirable. The search for new antiepileptic agents is reasonable in order to reduce the proportion of drug-resistant patients. The choice of conventional or new drugs should include not only expected efficacy and risk of adverse effects, but also pharmacokinetic properties and expense. Consequently, no general rule is appropriate, and each decision and recommendation for treatment should be individualized. PMID- 7481301 TI - [Stabilization treatments of migraine]. AB - Pathophysiology and principles for diagnosis of migraine, in particular ways to obtain an accurate history between attacks, are presented in the introduction. Identification of migraine triggers is an extremely important part of migraine therapy. Selected pain-relieving drugs should include primarily simple analgesics and antiemetics, while sumatriptan should be reserved mainly for severe migraine attacks. Pharmacotherapy, psychological and physical therapy are all components of a systemic approach to the treatment of migraine. To discontinue overused medication is the first step in the prevention of migraine. A number of available drugs are able to reduce the frequency and severity of migraine attacks. Several considerations should be evaluated before symptomatic and prophylactic therapies are introduced. Our strategies are based on experience collected during the treatment of 82 patients. PMID- 7481303 TI - [Myasthenia-like syndromes: current and future treatments]. AB - Therapeutical aspects of disorders of the neuromuscular transmission are based on differential diagnosis between pre- or post-synaptically localized abnormalities of the neuromuscular junction, recognition of acquired and congenital forms and search for associated diseases and tumors, on prescription of drugs improving the transmission of acetylcholine and immunosuppressive agents, and on regular and quantified follow-up. In this clinical and therapeutic review, the two most common forms, myasthenia gravis and the Lambert-Eaton myasthenic syndrome, are discussed in the light of the 44 patients recently seen in our department. PMID- 7481306 TI - [What is your diagnosis? Coronary heart disease]. PMID- 7481305 TI - [Antisense oligonucleotides--a new generation of potential drugs]. PMID- 7481304 TI - [Toward a treatment for dementia]. AB - The clinical manifestations of so-called 'untreatable dementias' result from neuronal dysfunction causing premature neuronal death. As long as the neurosciences won't have an explanation for the increased vulnerability and cause of death of neurons in neurodegenerative disorders, no preventive or curative treatment can be expected. So far, the treatment of dementia focuses essentially on the consequences of neuronal dysfunction or cell loss by either a palliative approach addressing psychosis, behavior or anxiety and depression, or by substitution of deficient neurotransmitters with quasi no success. Only tacrine, a cholinesterase inhibitor, now available in Switzerland, has a marginal effect in early cases, but every other substitutive approach has failed so far. Muscarinic agonists as well as antiamyloid substances will be tested soon in clinical trials. Growth factors (especially NGF) raise big hopes for the near future, but they are still under preclinical evaluation. PMID- 7481307 TI - [Drug treatment of benign prostatic hyperplasia]. PMID- 7481309 TI - [Rare association of a coronary fistula with a fusiform aneurysm of the superior vena cava: diagnosis via transesophageal echocardiography and MRI]. AB - A congenital fistula of the left circumflex coronary artery with large aneurysmal sacculations and drainage into the vena cava superior is reported in an asymptomatic black adult female. An indicator dilution curve excluded a significant left-to-right shunt. In addition, the patient had a large fusiform aneurysm of the superior vena cava with maximal extension in the anterior upper mediastinum. Transesophageal Doppler echocardiography and magnetic resonance imaging were complementary diagnostic tools, the first for clearly visualizing coronary anatomy and shunt, the second for accurate imaging of the aneurysmal vena cava superior in the upper mediastinum. PMID- 7481308 TI - [Germ cell tumors in children and adolescents]. AB - 3.7% of the diagnoses in the Pediatric Tumour Registry (Zentrales Tumor-Register) of the German Society for Pediatric Oncology and Haematology (Gesellschaft fur Padiatrische Onkologie und Hamatologie) concern germ cell tumors. The annual incidence has increased from 0.22 in 1980 to 0.60 per 100,000 children in 1992. The most common localizations are the coccygeal region (25%), followed by the testes (20%), the ovaries (20%) or the brain (20%). Some malignant germ cell tumors tend to secret the tumour markers Alpha-1-Feto-protein and HCG, and may then be diagnosed clinically due to the increased serum or spinal fluid levels. In the cases of extensive tumors growth and secreting intracranial germ cell tumors, a preoperative chemotherapy is favourable. The latest chemotherapy regime containing Cis-Platinum allows us to achieve long term remission in 80% of cases. Risk factors for the regime include histology, localization stage, and incomplete resection of the primary tumors. PMID- 7481310 TI - [Pneumonia, biclonal gammopathy, paralysis of the fibular nerve and cerebrovascular insult]. PMID- 7481311 TI - [Recurrent respiratory tract infections]. AB - On the occasion of a treatment of frequent infections of the upper respiratory tract an antibody deficiency syndrome was noticed in a 32-year-old female patient. Further investigations revealed a variable immunodeficiency syndrome, a disease belonging to the primary humoral immune defects and presenting with multiple clinical manifestations. Leading symptoms, however, are recurring respiratory infections that may lead to bronchiectasis. Therapy consists in the prevention of infections by periodical intravenous substitution with 7-S immunoglobulins and opportune prescription of antibiotics. PMID- 7481312 TI - [A case from practice (333). Amebic abscess of the left liver lobe. Left-sided kidney cyst]. PMID- 7481313 TI - [What is your diagnosis? Proctalgia fugax]. PMID- 7481314 TI - [Tuberculosis in the next century?]. PMID- 7481315 TI - [Air pollution in Switzerland and respiratory diseases in adults.Results of a preliminary study of the cross-sectional part of the Sapaldia study]. AB - In the cross-sectional part of the Sapaldia study, 9651 participants answered a health questionnaire and were submitted to medical tests. In order to look for possible environmental influences from the environment, urban, rural and alpine regions were selected. As expected, the highest concentrations of NO2, SO2 and particles were measured in cities. The study showed that the prevalence of respiratory symptoms or diseases was higher in the more polluted areas. Forced vital capacity was lower in regions with higher mean annual values of pollutants, as compared with regions with the lowest mean values. Allergology tests showed that about one third of the population was atopic. Passive exposure to tobacco smoke, especially at the workplace, was associated with increased risk and prevalence of respiratory symptoms and diseases, such as asthma, bronchitis and chronic bronchitis. In conclusion, long-term exposure to environmental pollutants, even at low concentrations, significantly decreases the lung-function and increases the risk for respiratory symptoms or diseases. PMID- 7481316 TI - [Lung problems in acute to subacute exposure to medium altitudes]. AB - The hypobaric hypoxia of moderate altitude elicits various mechanisms of acute to subacute physiologic adaptations of the healthy lung: First of all it causes a hyperventilation, which increases the diminished arterial pO2. Because of hypoxemia-induced vasoconstriction, pulmonary arterial hypertension develops. The adrenergic stimulation of the cardiac output also increases the pulmonary perfusion. Most likely because of the diminished density of ambient air there is a measurable increase of exspiratory bronchial flow or, respectively, a diminution of the peripheral airway resistance. In higher altitudes, limitation of oxygen-diffusion under physical exertion is observed. The consequences of acute hypobaric hypoxia for diseased lungs depend on preexisting ventilation/perfusion mismatch or diffusion impairment. Arterial hypoxemia and hence also pulmonary arterial hypertension are increased. In the presence of normal chemoreceptor sensitivity (type pink puffer), a hyperventilation, which is often perceived as dyspnea by the patient, is induced. Mostly patients with chronic obstructive pulmonary disease adapt, however, surprisingly well to moderate altitude. Bronchial asthma improves frequently because allergen concentrations are low and air density is diminished. On the other hand, physical exertion in dry and cold ambient air may also elicit acute asthmatic exacerbations. The assessment of moderate altitude tolerance by patients with chronic lung diseases and prophylactic precautions before the ascent are discussed. The only altitude-specific disease of the healthy lung is the so called high-altitude pulmonary edema. The major pathogenetic factor for its development is an inadequate or overshooting response to hypobaric hypoxia (nonuniform pulmonary arterial vasoconstriction, diminished hypoxic ventilatory drive, retention of fluid, centralization of blood volume and capillary leak). Prophylactic and therapeutic implications are discussed. PMID- 7481318 TI - [Tuberculosis in Switzerland]. AB - In Switzerland, in 1992, 957 persons suffered from tuberculosis; 52.3% were Swiss, 47.7% foreigners. Most of the swiss TB patients were more than 65 years old, whereas the foreigners generally were young patients originating from countries with high TB-infection rates. Asylum seekers had much higher TB-case rates (131 cases per 100,000) than other foreigners (27 cases per 100,000) or Swiss (9 cases per 100,000). Special refuge reception centers have been set up in Switzerland, in charge of tuberculosis screening procedures in this high-risk group on arrival to this country. Although HIV and AIDS patients bear a much higher risk of developing tuberculosis once infected, the HIV epidemic did not lead to an increase of tuberculosis in Switzerland so far, since young Swiss are rarely infected with tuberculosis. HIV-infected, drug addicts, homeless persons and alcoholics run a higher risk of contracting tuberculosis only when congregating with a person suffering from active tuberculosis not yet diagnosed or improperly treated. In order to maintain low levels of tuberculosis in Switzerland DOT (directly observed therapy) must be implemented in all patients with uncertain compliance, especially as cultural and social backgrounds become increasingly complex. PMID- 7481317 TI - [Safety and side effects of inhalant steroid therapy in children with bronchial asthma]. AB - Therapy of asthma with inhaled steroids in children becomes the main standard for treatment of asthmatic airway inflammation. Thus, in most children asthma is controlled by the new galenic formulations of topical inhalative corticosteroids. Dose-dependent side effects must be known and have to be detected by appropriate testing. In childhood the suppression of the hypothalamic-pituitary adrenocortical axis and of growth is the main concern of steroid therapy. The latter depends on dose and duration of intake as well as the susceptibility of the growth phase during which the child inhales steroids. Today, doses of topical steroid up to about 800 micrograms/day seem to be safe. However, some cases with individual reactions to low doses of inhaled steroids are known: therefore, close clinical control of children on inhaled steroids is extremely important. PMID- 7481319 TI - [Therapy-resistant asthma: causes and therapy]. AB - Despite the introduction of new and potent antiasthmatic drugs, a minority of asthma patients, remains without response to therapy. Some of the patients with asthma refractory to therapy actually do not have asthma at all, but suffer from another syndrome mimicking asthma. Asthma may also exist in combination with a syndrome mimicking asthma (e.g. vocal cord dysfunction). Patients may then get treated with high doses of corticosteroids unnecessarily, because the symptoms due to vocal cord dysfunction are misinterpreted as being asthmatic. Sometimes asthma is complicated by an independent factor (e.g. gastroesophageal reflux), which does not respond to antiasthmatic drugs. In all cases of asthma being unresponsive to therapy, the diagnosis of asthma must be questioned, and other complicating factors need to be excluded. The patient's compliance should be encouraged, and correct handling of modern inhalation devices has to be monitored. Patients with refractory asthma should also undergo flexible bronchoscopy to exclude morphologic changes of the bronchial system and to obtain mucosal biopsies. PMID- 7481320 TI - [Therapeutic problems in pneumonia]. AB - In Switzerland 10 out of 1000 adults suffer from pneumonia each year. It is of note that mycoplasma, influenza virus and pneumococci are the most common causative agents of community-acquired pneumonia. For the latter macrolides are presently the antibiotics of choice. Pneumonias occurring in patients with immune disorders should be treated primarily with amoxicillin + clavulanic acid or with cephalosporins of the second generation, because infestation with germs like haemophilus influenzae and klebsiella pneumoniae have to be considered. If the empirically chosen therapy should fail, the therapeutic strategy should not be changed blindly. Differential diagnosis and appropriate investigations are necessary (other germ, other disease, complications?). Problems in treating patients with pneumonia are illustrated by three case examples. PMID- 7481321 TI - [What is your diagnosis? Scimitar variant (abnormal right-sided pulmonary veins with normal outlet in left atrium) with normal size right lung]. PMID- 7481322 TI - [Stress and immune function: facts and myths]. PMID- 7481323 TI - [Current therapy concepts in early and intermediate stages of Hodgkin diseases in adults]. AB - 80% or more of patients suffering from Hodgkin's lymphoma (HD) of limited stage are cured with modern treatment. In current scientific studies a controlled and stepwise reduction of the treatment intensity in early stage HD is investigated. Overall survival rate 20 years after treatment is approximately 65%. This figure is approximately between 15 and 20% lower than expected according to the rate of tumor control. This discrepancy is due to an increased incidence of certain other diseases, with an increased incidence of secondary solid malignancies as the main problem. We don't know the relative importance of disease-associated factors, hereditary factors or of treatment-associated factors for this increased incidence of secondary solid malignancies. Irradiation of large volumes is considered an important risk factor. Several retrospective analyses show that the incidence of secondary solid malignancies is significantly lower after limited irradiation compared to large field irradiation. The incidence of secondary solid malignancies after combined treatment with limited field irradiation plus chemotherapy is not higher when compared with chemotherapy alone. After chemotherapy alone this risk is comparable or less than after large field irradiation alone. Finally there is only a small risk of leukemia after treatment with ABVD. An influence of diagnostic laparotomy with splenectomy on the survival of these patients has not been observed. Several retrospective analyses and at least one randomized study with limited chemotherapy, combined with limited irradiation, showed a tumor control rate and an overall survival rate comparable to the figures after more intensive treatment. Based on clinical examinations, patients with CS I and II HD are grouped in low, intermediate and high risk levels.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7481324 TI - [The role of radiotherapy in the local treatment of Ewing's sarcoma]. AB - Therapy of Ewing's sarcoma consists of adequate local treatment for control of the primary tumor (surgery or radiotherapy or a combination of both) and systemic therapy. Tumors with favorable prognosis (small tumor volume, good response to chemotherapy) may be managed by a single local modality. In tumors with poor prognosis, combined local treatment (surgery plus radiotherapy) seems to improve local control and survival. As an innovative approach, preoperative radiotherapy for high-risk tumors is investigated in the EICESS-92-study. PMID- 7481325 TI - [Neuroblastoma: diagnosis and therapy]. AB - Prognosis of neuroblastoma is primarily dependent on the stage of disease. While stages 1 to 3 show a survival rate of 94 to 66%, stage 4 has a survival rate below 20%. The most important prognostic factor is the extent of disease. Age, LDH, resectability of primary tumor, cytologic and molecular parameters are defined to have a prognostic impact as well. A stage- and risk-adapted therapy of neuroblastoma needs a thorough assessment of all these factors. PMID- 7481326 TI - [Demonstration of mesangial IgA deposits in kidney biopsies of pediatric patients: comparison with the clinical picture]. AB - Clinical and morphological findings were evaluated in 25 children with mesangial IgA deposits. 19 patients had recurrent macroscopic hematuria (n = 10), chronic proteinuria > 40 mg/(m2.h) (n = 5), recurrent hematuria with chronic proteinuria (n = 3), or chronic nephrotic syndrome (n = 1). The glomerular involvement was similar in six patients with history of Schonlein-Henoch purpura and in 13 patients without such history: normal or nearly normal glomeruli (n = 3), focal and segmental glomerulonephritis (n = 11) and diffuse proliferative glomerulonephritis (n = 5). End-stage renal disease developed in two patients with proteinuria > 40 mg/(m2.h) and more than 50% of their glomeruli are affected by crescents. The common histopathological features in patients with and without history of Schonlein-Henoch purpura suggest a common pathogenesis. The risk of poor outcome appears, related to the severity of proteinuria and to the presence of crescents, in more than 50% of glomeruli. Mesangial IgA deposits were also demonstrated in six children with steroid-responsive idiopathic nephrotic syndrome: light microscopic studies revealed normal or nearly normal glomeruli in five and focal segmental glomerular sclerosis in one patient. The microscopic findings were clearly different in the six patients with idiopathic nephrotic syndrome as compared with the patient with chronic nephrotic syndrome, who presented with severe glomerular lesions and extensive crescent formation. The results indicate that the presence of mesangial IgA deposits in the clinical setting of idiopathic childhood nephrotic syndrome is incidental and that such patients should still be considered as having idiopathic nephrotic syndrome in spite of their immunopathological features. PMID- 7481327 TI - [Radiological case of the week (1). Pulmonary lymphangioleiomyomatosis]. PMID- 7481328 TI - [A case from practice (334). Hyperhidrosis of 1 1/2 year duration, probably postmenopausal. Arterial hypertension for years. Obesity]. PMID- 7481329 TI - [What is your diagnosis? Digitalis poisoning]. PMID- 7481330 TI - [Anatomic-pathological bases of coronary heart disease]. AB - Coronary artery disease is described by its extreme presentations. At one end we see the slowly growing fibrous plaque gradually occluding the artery. Collaterals develop and confine the loss of myocardium to the inner layer of the left ventricle. At the other end of the spectrum we see the rupture of a soft plaque. Blood breaking in an atheroma occludes a coronary artery suddenly. There is no time to develop collaterals. The myocardial infarction corresponds in its size to the perfusion-area of the occluded artery. Transmural infarctions lead to rupture of the free wall, ventricular septum or papillary muscle. The coronary morphology of our patients suffering from coronary heart disease lies between these two extremes. However, many patients die suddenly due to arrhythmia a long time before the loss of myocardium disturbs left ventricular pump function so severely that lethal heart failure may develop. PMID- 7481331 TI - [Asymptomatic ischemia--an important part of the spectrum of coronary disease]. AB - Angina pectoris and asymptomatic myocardial ischemia are part of the spectrum of coronary heart disease. Not the presence or absence of angina determines the future of the patient, but repeated ischemia and the progression of the coronaropathy. This progression is neither linear with time, nor is the moment of plaque rupture foreseeable. Silent myocardial infarctions increase with age and are very frequent in diabetics. In patients without neuropathy but with asymptomatic myocardial ischemia the central pain threshold is higher than in patients with angina pectoris. The best noninvasive test for the detection, localization and estimation of extension of myocardial ischemia, be it pain-free or symptomatic, is 201-thallium scintigraphy, combined with the exercise ECG. The fight against all amendable cardiovascular risk factors and pharmacotherapy are the first steps, if asymptomatic myocardial ischemia is suspected. Augmented dyspnea on effort and rhythm disturbances are indicators of advanced multivessel heart disease. Under these circumstances coronary angiography is indicated, and further treatment should follow the generally accepted rules such as for patients with angina pectoris. PMID- 7481332 TI - [Drug therapy in angina pectoris]. AB - The currently available antianginal drugs act by reduction of myocardial O2 requirement and/or by coronary vasodilatation. The choice between beta blockers, nitrates, calcium antagonists or their combination depends on the clinical presentation, coexisting disorders and specific factors in individual patients. In addition to symptomatic treatment, secondary prophylactic measures, such as aspirin and reduction of serum cholesterol, are also necessary to prevent progression of the underlying coronary artery disease. In this paper the comparative efficacy and the indications of the various types of antianginal drugs are discussed. PMID- 7481333 TI - [Interventional cardiology]. AB - The evolution of cardiology over the past 25 years has been dominated by the invention and development of interventional techniques. These techniques offer 'surgical treatment' through transvascular access to the heart, avoiding therefore the classic transthoracic surgical approach. Today, coronary artery disease, valvular heart disease and arrhythmias are accessible to interventional treatments. In this article, a review is given about recent developments and results of interventional treatment for coronary artery disease with special consideration of stenting. This article should help the general practitioner or internist to guide and inform their patients suffering from coronary artery disease. PMID- 7481334 TI - [Thrombolysis of myocardial infarction--state of the art]. AB - Thrombolytic therapy has revolutionized the treatment of acute myocardial infarction by reducing mortality and preserving left ventricular function. It is a relatively safe and cost-effective therapy, although underused in many countries. To further improve the results and the availability of this therapy, the reduction of the door-to-needle time in hospitals is the most important strategy. Alternatively, prehospital thrombolytic therapy has been shown to be feasible and to result in a similar risk reduction as thrombolytic therapy started in hospitals. In this overview the results of recent studies are summarized and discussed in view of defining indications more appropriately. Our current knowledge of adjunctive therapy is summarized briefly, and the benefits, risks and costs of thrombolytic therapy are presented. Interventional procedures are emphasized for treatment of patients with cardiogenic shock and unstable angina. Besides emergency and rescue PTCA, interventional therapy is an attractive and superior alternative to thrombolytic therapy. However, the results of interventional strategies depend largely on the operators and institutional experience and structural facilities, which are limited to a small number of institutions. Thus, any attempt has to be undertaken to provide thrombolytic therapy as early as possible to a patient population as large as possible. PMID- 7481336 TI - [What is your diagnosis? Broad complex tachycardia]. PMID- 7481335 TI - [The other heart--pictures and thoughts for a farewell]. AB - This is an attempt to draw attention to the demanding and complementary reality of modern medical doctors by confronting cardiology and cordiology (symbolism and mythology of the heart). After a survey of the symbolic and mythological world of the heart, the question of compatibility between the apparently opposing poles of cardiologic curative technology and cardiologic emotionalism is posed. With respect to a comprehensive medicine of tomorrow, it is crucial whether modern doctors will be capable of a two-fold synthesis, namely the harmonious interaction between a rational basic position (raison de la mathematique) and an emotional standpoint (raison du coeur) and the increasing harmony of male and female norms. PMID- 7481337 TI - [Cardiotoxicity and cardioprotection in chemo- and radiotherapy for malignant diseases]. PMID- 7481338 TI - [Cardioprotection in chemo- and radiotherapy for malignant diseases--an echocardiographic pilot study]. AB - AIM: Pilot study, examining the cardioprotective effect of an antioxidant regimen in patients with malignancies receiving high dose chemo- or radiotherapy. PATIENTS AND METHODS: 14 patients with chemotherapy and 10 patients with radiotherapy were randomized in a double-blind fashion (placebo versus vitamin E and C and N-acetylcysteine). Systolic and diastolic echocardiographic parameters were determined before and within three weeks of treatment completion. RESULTS: Left ventricular ejection fraction fell significantly in patients receiving placebo (radiotherapy: 67 +/- 6 to 56 +/- 2%, p = 0.008, chemotherapy: 67 +/- 7 to 60 +/- 8%, p = 0.05). Patients on antioxidants showed no significant fall in EF (radiotherapy: 63 +/- 8 to 61 +/- 7%, chemotherapy: 67 +/- 5 to 64 +/- 6%). CONCLUSION: The small number of patients in the study precludes a definitive statement. The preliminary results, however, suggest efficient cardioprotection by this cheap and safe antioxidant combination, so that larger studies are warranted for confirmation. PMID- 7481341 TI - [Benefits of a new galenic form of levodopa and benserazide in the treatment of patients with Parkinson disease]. AB - We describe a new, water-soluble formulation of levodopa plus benserazide (Madopar LIQ) for the treatment of Parkinson's disease. Two dosage strengths are available: 100 mg levodopa plus 25 mg benserazide and 50 mg levodopa plus 12.5 mg benserazide. Pharmacokinetic data show that levodopa absorption is more rapid than with standard Madopar, resulting in a shorter time to peak plasma concentration. Other pharmacokinetic values are comparable to those obtained with the standard formulation. We discuss the clinical advantages of this new water soluble formulation, particularly when the patient requires rapid onset of action for morning or circadian akinesia. The indications of this formulation in patients with dysphagia and in other clinical situations, e.g. during the postoperative period and for levodopa dosage adjustment in ambulatory care, are discussed. PMID- 7481340 TI - [Multiple familial pheochromocytomas: sonographic demonstration of multiple adrenal, celiac and bladder localizations in a child]. AB - Familial pheochromocytoma is an uncommon form of this neoplasia. It is characterized by an autosomal dominant inheritance and multicentric locations. It is more frequently encountered in children, but with a smaller risk of malignant transformation than in adults. The familial form may be associated with other disorders, particularly with multiple endocrine neoplasia (MEN), Von Hippel Lindau's disease and Von Recklinghausen's disease. We report the case of a nine year-old boy with multiple familial pheochromocytoma in whom sonography allowed to demonstrate five localizations. PMID- 7481339 TI - [Prevalence of medullary sponge kidney in patients with and without nephrolithiasis]. AB - Reports on the prevalence of medullary sponge kidneys have given conflicting results. The present work is aimed at defining the prevalence of this abnormality in renal stone formers compared with a non-stone-forming control population by strict radiological criteria. Three separate studies have been carried out: the retrospective analysis of 191 intravenous urographies, which had been performed in 1988; the respective prospective analysis of 104 consecutive intravenous urographies and of 149 consecutive urographies, which had been performed at the end of intravenous digitalized subtraction renal angiographies. The 444 patients were classified according to presence or absence of nephrolithiasis. After exclusion of 70 patients (16%) of which the X-ray didn't fulfill the quality criteria, urographies were classified as 'medullary sponge kidneys', 'papillary blush' or 'negative' according to the radiological aspect of the papillae. Diagnosis of medullary sponge kidneys was based on presence of at least three linear or round papillary opacities seen in at least on papilla even of one kidney on late films (at least ten minutes after injection of contrast medium), taken without ureteral obstruction or abdominal compression. Overall results reveal that prevalence of a medullary sponge kidney in renal stone formers (8.5%) is significantly (p < 0.01) higher than in the control population (1.5%) and higher than what has been generally reported. Results of the retrospective and of both prospective analysis did not significantly differ. The best diagnostic tool to unmask medullary sponge kidneys remains urography. PMID- 7481342 TI - [Radiological case of the week (2). Intestinal wall gangrene in mesenterial infarction with intramural and intraportal gas]. PMID- 7481343 TI - [A case from practice (335). Stage B3 HIV infection. Acute cholecystitis with cough. Pruriginous dermatitis. Chronic hepatitis B]. PMID- 7481344 TI - [Chest pain, dyspnea, syncope]. PMID- 7481345 TI - [Diabetes]. PMID- 7481346 TI - [Behavior of pancreatic beta-cells in type I and II diabetes: clinical implications?]. PMID- 7481347 TI - [Therapeutic approach to a multi-metabolic syndrome]. PMID- 7481349 TI - [Gestational diabetes]. PMID- 7481348 TI - [Genetics of type II diabetes: current status!]. PMID- 7481350 TI - [Quality of blood sugar control and late complications of diabetes]. PMID- 7481351 TI - [Diabetes and infection]. PMID- 7481352 TI - [AIDS and diabetes]. PMID- 7481353 TI - [Inefficacy of oral antidiabetics: failure or natural history of diabetes?]. PMID- 7481354 TI - [Type II diabetes (NIDDM: non-insulin dependent diabetes): they must be diagnosed, they must be treated; it is urgent!]. PMID- 7481355 TI - [Management of the insulin-dependent diabetic in ambulatory practice: the first year]. PMID- 7481356 TI - [Management of type II diabetics in ambulatory practice]. PMID- 7481357 TI - [The pancreatic heart]. PMID- 7481358 TI - [Alcohol and diabetes]. PMID- 7481359 TI - [Insulin therapy today and tomorrow]. PMID- 7481360 TI - [The kidney in diabetes]. PMID- 7481361 TI - [Diabetes secondary to endocrine diseases]. PMID- 7481362 TI - [Diabetes and pancreatic tumors]. PMID- 7481363 TI - [Treating diabetic patients: various personal experiences]. PMID- 7481364 TI - Lasers in surgical oncology: Part I. PMID- 7481365 TI - Laser photochemotherapy: a less invasive approach for treatment of cancer. AB - The effectiveness of combining surgery with chemo- and radiation therapy in treatment of human cancer provides a useful model for further development of new multimodality approaches including laser photochemotherapy. Laser endoscopy often is a useful treatment for obstructive tumors in airways, but interstitial laser fiberoptics is becoming a more precise, minimally invasive alternative for ablation of unresectable or recurrent neoplasms. Combining intratumor chemotherapy with laser energy delivery via interstitial fiberoptics should be most effective using drugs activated by photothermal energy. A number of investigators have shown that anthracyclines and cis-platinum are likely candidates for light or heat activation in cancer cells. An advantage of anthracyclines is their dual role as antitumor drugs and as photosensitizers. Because they are effective chemotherapy agents without photoactivation, two approaches are possible to increase tumor responses. Maximum tolerated dose followed by photoillumination via laser fiberoptics can be used to obtain better tumor palliation. Improved treatment response to lower intratumor drug levels after laser activation also should reduce systemic toxicity. Preclinical studies and recent case reports from several groups suggest photochemotherapy with currently approved drugs and lasers may soon become an attractive alternative for treatment of recurrent tumors in cancer patients. PMID- 7481366 TI - Nd:YAG laser procedures in tumor treatment. AB - Due to the wide variability of tissue interactions and the possibility of specific applications, the neodymium-yttrium-aluminum-garnet (Nd:YAG) laser is the most important surgical laser. With the adequate choice of application mode and relationship between interaction time and power density, it can be used for precise cutting in the contact mode with the bare fiber, with either a wide or small coagulation seam. With a handpiece a precise focal coagulation for preparation and hemostatic purposes is possible, as well as cutting with the focussed noncontact beam where additionally a wide coagulation seam is necessary. Endoscopically guided coagulation, vaporization, and cutting are possible with bare fibers which can be introduced through nearly all endoscopes. With increasing importance, the possibility for wide and homogeneous volume coagulation is used to destroy diseased tissues either by noncontact irradiation or interstitial placement of the fiber. Thus, the field of laser application in tumor therapy ranges from the treatment of superficial tumors to endoscopic tumor ablation, resection of neoplastic tissue in parenchymatous organs, and interstitial thermotherapy with coagulation of deep-seated primary and secondary malignancies. With its different application modes the laser can be used as a surgical instrument or as a central therapeutical method, whereby perfect control of tissue interactions is always possible by using either visual control for superficial and endoscopic procedures or magnetic resonance imaging and color coded duplex sonography as a control for interstitial procedures. During 12 years of clinical work we have developed several application modes and have proved the Nd:YAG laser to be an effective instrument in tumor therapy. PMID- 7481367 TI - Laser surgery in gynecologic oncology. AB - In the treatment of premalignant epithelial cancers of the female lower genital tract, the CO2 laser beam is used with precision through a surgical microscope for tissue ablation and excision. Intra-abdominal vaporization of abnormal tissues can be performed endoscopically through fiberoptics. Surgical lasers provide thermal scalpels for hemostatic excision of malignant lesions. Photodynamic therapy is applied in the treatment of isolated malignancies persistent after conservative treatment. Surgical laser systems are important tools in the treatment of gynecologic malignancies. PMID- 7481368 TI - Endoscopic treatment of gastrointestinal tumors: indications and results of laser photocoagulation and photodynamic therapy. AB - Both Nd:YAG laser and photodynamic therapy (PDT) have ideal indications in endoscopic treatment of gastrointestinal (GI) tumors. A retrospective analysis of 3,505 Nd:YAG laser treatments in 1,015 patients revealed that recanalization of inoperable tumors can be obtained in 93% and 97% of patients with upper and lower GI neoplasms, respectively. Subjective improvement of symptoms was achieved in 74% and 97%, respectively. The overall morbidity and mortality rates were 3% and 1%, respectively, for upper GI tumors and 3% and 0.5% for lower GI tumors. Colorectal adenomas were eradicated in 84% of cases, with a morbidity and mortality of 5% and 0%, respectively. Early stage esophageal and gastric carcinomas were treated with Nd:YAG laser when lesions had well-defined borders or protruded over the mucosal surface and with PDT (38 PDT cycles in 27 patients) in the case of undefined borders or ulcerated lesions. The cure rate was 73% for esophageal tumors and 85% for gastric tumors. Sunburns after photosensitization and local complications occurred in 7% and 6% of patients, respectively. No death was related to endoscopic treatment. PMID- 7481372 TI - [Fatal familial insomnia and prion diseases]. AB - Fatal familial insomnia has recently enlarged the group of prion diseases. The disease starts between 35 and 60 years of age, is inherited as an autosomic dominant trait, and leads to death within 7 to 32 months. Clinical symptoms and signs include insomnia dysautonomia, cognitive and motor alteration. The discrete topography of the lesions in fatal familial insomnia underlines the role of the thalamus in the regulation of the sleep-wake cycle. Atrophy, neuronal loss and gliosis are prominent in the anterior and dorsomedial nuclei of the thalamus. Spongiosis, which is usually found in prion diseases, is absent in fatal familial insomnia. An abnormal prion protein (PrPsc) is detected in the brain. There is a mutation at codon 178 of the gene encoding this protein. Fatal insomnia is distinct from Creutzfeldt-Jakob disease on clinical, histopathologic and molecular grounds. It provides new information about genetics of prion diseases which share the characteristics of being altogether inherited and, in most cases, transmissible. The recent finding of abnormal PrP in diffuse subcortical gliosis suggests that other degenerative disorders could actually be prion diseases. PMID- 7481371 TI - [Is there a single form of Charcot-Marie-Tooth disease, or are there several variations? A classification difficulty]. PMID- 7481369 TI - Lasers in rectosigmoid tumors. AB - Over a 14-year period, 746 patients were treated at the Lille Laser Center for rectosigmoid tumor. Two hundred seventy-two patients were treated for palliation of symptoms from rectosigmoid cancer and 474 patients were treated for a benign rectosigmoid villous adenoma. The immediate success rate and complication rate were 85% and 2%, respectively, for patients with advanced cancer, and 92.8% and 1.8% for those with a villous adenoma. Patients with an advanced cancer remained functionally improved during a 10.1-month average period after initial improvement. The recurrence rate after initial treatment for villous adenomas was 18% during a 29.7-month average follow-up. Immediate results were influenced by reason for treatment, initial symptoms, and circumferential extension for patients with a cancer, and only by circumferential extension for patients with a villous adenoma. Long-term results were influenced by reason for treatment and circumferential extension for patients with cancer, and by reason for treatment, initial histology and localization for patients with a villous adenoma. Laser treatment provided relief of symptoms in inoperable patients with advanced rectosigmoid carcinomas. Because treatment is long and difficult and the cancer rate is high, endoscopic laser should be limited in patients with a circumferential villous adenoma to non-surgical candidates. The risk of complication after surgery (some being fatal) has to be balanced against the risk of undetected carcinoma and the indication for endoscopic laser treatment should be discussed case by case. PMID- 7481370 TI - Applications of laser technology in breast cancer therapy. AB - Breast cancer is an increasingly common problem affecting one in nine women. The optimal management of carcinoma of the breast remains controversial. This paper reviews the rationale for the use of laser technology in the treatment of primary and advanced breast cancer. The CO2 laser has several properties which make it advantageous for breast surgery. The technical details for optimal laser utilization are presented. Experimental evidence documents a marked reduction of local tumor recurrence following surgery with lasers. Preliminary human studies suggest that laser use lengthens the disease-free interval and may decrease local recurrence. Interstitial laser therapy holds promise for use in the treatment of locally advanced breast tumors and has been suggested by some as a potential modality for the primary therapy of breast cancer. The clinical use of lasers in the treatment of breast cancer is justified. PMID- 7481374 TI - [Progressive severity of left unilateral apraxia in 2 cases of Alzheimer disease]. AB - Two patients presented with progressive left unilateral motor apraxia and progressive visuo-spatial difficulties, including constructional apraxia, dressing apraxia, spatial dysgraphia and dyslexia, spatial acalculia and neglect of the left side, without significant changes in the other cognitive functions. In both patients, radiological tests demonstrated cortical atrophy, more marked in the retrorolandic areas. A diagnosis of Alzheimer's disease was made in the first patient by cortical biopsy and in the second patient by post-mortem examination. The second patient died from an intercurrent neoplastic condition early in the course. Post-mortem examination of her brain showed the atrophy to be predominantly in the superior parietal gyri. In these areas, in addition to neuritic plaques and neurofibrillary tangles, these was also severe gliosis and superficial spongiosis. These two cases were clinically different from cortico basal degeneration, where the motor difficulties are associated with occulo-motor problems, extrapyramidal signs and involuntary movements of the "alien hand" type. These data confirm the presence of focal forms of Alzheimer's disease. The motor apraxia seen in our patients resembles the kinesthesic apraxia described by Luria. PMID- 7481373 TI - [Isoform 4 of apolipoprotein E and Alzheimer disease. Specificity and clinical study]. AB - We studied apolipoprotein E (apoE) phenotype in 113 patients with possible and probable Alzheimer's disease (AD), 49 patients with Parkinson's disease (including 11 patients with dementia) and 23 patients with mixed and vascular dementia. Normal controls were 498 young, healthy blood donors previously recorded. All patients were assayed for blood lipid parameters. All AD patients underwent a neuropsychological evaluation (including a mini-mental status and 5 subtests of Cole and Dastoor hierarchic dementia scale) and a detailed interrogation of them and their caregivers about their familial and personal medical history. The recorded data included age at onset, clinical subtype (i.e. amnesic or aphaso-apraxic), occurrence of fits, cases of probable dementia in relatives, and ages of their parents at death. There was a significant association between the fourth isoform of apoE and AD, as in previous works. We did not found such an association for PD patients (even with dementia) nor mixed and vascular demented patients. We failed to find any association between any clinical characteristic of the patients and the biological subgroups defined by the number of epsilon 4 alleles, except with regard for the age of onset. Surprisingly, the mothers of epsilon 4 bearers had a significantly longer life than mothers of other patients. We failed to found any significant difference of apoE2 isoform frequency between AD patients and controls. AD patients had higher levels of cholesterol and apoAl than did MP and mixed and vascular demented patients. ApoAl level is known to constitute a protective factor against coronary heart disease, which is usually increased by the presence of apoE-epsilon 4.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7481375 TI - [Anarthria, progressive apraxia and extrapyramidal syndrome: an uncommon clinical form of corticobasal degeneration? A case studied by HMPAO Tc99m single-photon emission tomography]. AB - A 48-year-old man presented with impaired joint movement and buccofacial apraxia. The disease progressed for six years associating an akineto-hypertonic syndrome, marked anarthria, saliva incontinence, bi-opercular syndrome, bucco-facial apraxia, severe global gestual apraxia and a frontal syndrom. Oculo-motricity and gait were normal. Magnetic resonance imaging of the brain demonstrated fronto parietal atrophy and HMPAO Tc99 tomography revealed hypoperfusion of the cortex clearly predominating in the left parietal region. These particular findings with predominantly intense joint involvement is similar to the clinical picture in corticobasal degeneration--subcortical signs (progressively uncontrollable hypertonia) together with cortical signs (severe gestual apraxia). The neuroradiological imaging and functional results also suggest a degenerative process. PMID- 7481377 TI - [Isolated angiitis of the central nervous system. Clinical and neuropathological study of 2 cases]. AB - Isolated angiitis of the central nervous system (IAC) is an idiopatic, recurrent vasculitis confined to the CNS involving small blood vessels. We describe the clinical, angiographic, and neuropathological data in two patients with IAC and delineate the main clinical and neuropathological features in both cases as well as the importance of a complete autopsy for discovering subclinical vasculitic lesions outside the CNS. Patient 1 concerned a 40 year-old-man that evolved for the last three years, initially with focal seizures, headache, and neurological focal deficits, later on the left sided hemihyposthesia and preferentially left parieto-occipital dysfunctions. He presented an oligoclonal band in CSF with slight hyperproteinorraquia and 25 lymphocytes. A cerebral angiography was compatible with angiitis and a leptomeningeal/cerebral biopsy showed lymphocytic vasculitis in the leptomeningeal and intraparenchymatous cerebral small vessels. These results lead to start a treatment with Cyclophosphamide associated to high dose of steroids. The patient clearly improved and now is almost asymptomatic. Patient 2 concerned a 67 year-old-man that evolved for 4 years with encephalic ischemic lesions distributed and confined throughout the brain stem and cerebellum, temporary remissions occurred and the patient required high-dose steroids and Cyclophosphamide to improve. Conventional and MRI angiographies only suggested the diagnosis that was confirmed at autopsy. The patient died after a massive pulmonary thromboembolism and a complete necropsic study showed abundant lymphocytic infiltrates, without granulomatous lesions, in the intraparenchymatous and leptomeningeal cerebral small vessels specially at the brain stem and cerebellar level where many demyelinated greyish areas and few infarctions were to be seen. The inflammatory cells were, in both cases, predominantly CD4+ T lymphocytes.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7481376 TI - [Clozapine in the treatment of mental manifestations of Parkinson disease]. AB - BACKGROUND: Medical treatment of Parkinsonian syndromes is often complicated by psychiatric side effects such as confusional states, hallucinations and psychosis. Recent pilot studies report good clinical results with the atypical neuroleptic clozapine. PATIENTS AND METHODS: We report on 15 patients with Parkinsonian syndromes: 11 with idiopathic Parkinson's disease (IPD), 3 with multiple system atrophy (MSA) and 1 with postencephalitic Parkinsonism (SPP). The mean age was 68.8 +/- 10 years; the mean duration of Parkinsonian symptoms was 6.8 +/- 5.7 years. The Hoehn & Yahr grade was: 3.5 +/- 0.8. Eleven patients were suffering from psychotic episodes, 10 from hallucinations, 8 from confusional states. Clozpine was introduced at nighttime and dosage was modified until the appearance of clinical effect or intolerable side effects. RESULTS: We report on an observed cumulative duration of clozapine treatment of 13 patient-years. The average treatment duration was 10.5 +/- 10.4 months. The mean daily dose was 33.3 +/- 30 mg (range: 6.2-100). There was at least transitory improvement of psychiatric symptoms in all patients. There was constant and complete improvement in 7 patients (46%) and satisfactory improvement in 5 patients (33.3%). The levodopa dosage was unchanged (mean dosage 563 +/- 232 mg), and the dosage of dopamine agonists was significantly increased. None of our patients experienced motor deterioration. Side effects comprised sialorrhoea, sedation, orthostatic hypotension, and delirium tremens and an epileptic seizure in one patient each. Two patients died suddenly at the 63rd and at the 86th day of treatment respectively, outside the hospital. These deaths seemed to be unrelated to the treatment. There was no agranulocytosis. CONCLUSION: Clozapine is an efficient antipsychotic drug in Parkinsonian patients with no motor side effects in the dosages used. The effective dosage is very low in comparison to psychiatric patients. However various side effects may occur and close monitoring is required. PMID- 7481379 TI - [Loss of psychic auto-activation. Obsessive-compulsive behavior. Toxoplasmic abscess of the basal ganglia]. AB - A 34 year old woman with AIDS presented with adynamia and obsessive-compulsive behaviour. CT scan revealed bilateral toxoplasmic lesions involving basal ganglia. The neuropsychological picture resolved under specific antitoxoplasmic treatment. However, relapse of toxoplasmosis occurred in associating with the same behavioral changes. This is the first case of such a syndrome due to infectious lesions, and in which improvement through transient, was observed. PMID- 7481380 TI - [Claude syndrome caused by mesencephalic infarction: 2 cases]. AB - Paramedian midbrain infarcts limited to the oculomotor nerve fibers are uncommon. We studied 2 cases where the clinical syndrome included a third cranial nerve palsy and a contralateral cerebellar ataxia. The CT scan disclosed a paramedian midbrain tegmental infarct, so that it is possible to term our two cases Claude's syndrome. The oculomotor nerve fascicular palsies were complete in one case and limited to the extraocular muscles in the second case. The selective involvement of oculomotor function suggests intraaxial fascicular organisation of the third cranial nerve in the brainstem. PMID- 7481383 TI - [Benign neuropathy of the chin]. AB - A 49-year old man without relevant prior neurological disorder but with overweight, dyslipidaemia and mild glucose dysregulation, noticed on awakening a spontaneous, left-sided and painless numb chin, attributed to isolated mental neuropathy, which regressed entirely within a few weeks. The clinical condition was stable at 1-year follow-up. Ancillary investigations (including electromyography) did not disclose any cause to this episode, notably there was no underlying malignancy or systemic disease. A thorough literature review revealed another similar case which too remained unexplained. The hypothesis of a microvascular neuropathy in relation to vascular risk factors is cautiously raised; such a mechanism has been inferred previously in exceptional cases of regressive mental neuropathy in the context of sickle-cell crisis or systemic vasculitis. Thus, at variance with classical notions, the outlook of the numb chin syndrome is occasionally benign. PMID- 7481381 TI - [Cerebral infarction disclosing neurocysticercosis]. AB - An ischaemic stroke related to neurocysticercosis observed in the Paris neighbourhood is reported. A 32-year-old man originating from Cape Verde presented a left lacunar stroke. CT scan showed multiple areas of calcifications and a left capsulostriatal lucency. Gadolinium enhanced MRI disclosed significant abnormalities surrounding the left middle cerebral artery in the sylvian fissure. The titers of anticysticercian antibodies were highly positive in the serum and the CSF. The investigations in search of another cause of the stroke were negative. The outcome was good after a treatment associating albendazole and methylprednisolone. The increasingly recognized links between neurocysticercosis and stroke, especially in endemic areas are reviewed. Neurocysticercosis is an exceptional cause of ischaemic stroke in non endemic areas. In non-endemic areas, this cause of stroke has to be systematically suspected in patients coming from endemic areas. The diagnosis is based upon MRI, CT scanner and serologic tests. Treatment requires anti-parasitic drugs. The role of the corticosteroids is discussed. PMID- 7481384 TI - [Exacerbation of myasthenia gravis by pefloxacin]. AB - Pefloxacine 800 mg single dose was given as routine treatment after a cystomanometric examination to a 45-year-old woman with a 30-month history of generalized myasthenia gravis. One hour after, the patient developed exacerbation of myasthenia gravis with bilateral ptosis and an increased generalized weakness. She experienced a rapid improvement during the next 8 hours and physical examination returned to normal within one day. No additional factors which might have contributed to the exacerbation of myasthenia gravis were found. The report of exacerbation of myasthenia gravis with other antibiotic belonging to the group of fluoroquinolones (ciprofloxacin, norfloxacin and ofloxacin) prompt us to recommend caution with the use of all fluoroquinolones in myasthenic patients. PMID- 7481382 TI - [Narcolepsy disclosing neurosarcoidosis]. AB - A 37-year-old man developed excessive daytime sleepiness, sleep attacks and cataplexy revealing an hypothalamic tumour. Multiple Sleep Latency Tests (MSLT) were characteristics of narcolepsy. Tissue typing was positive for HLA DR2 and DQ1. Most cases of narcolepsy are idiopathic without any evidence of brain pathology. Although symptomatic narcolepsy may occur occasionally with diencephalic lesions. The relationship between narcolepsy with diencephalic lesions is unsettled and will be discussed. PMID- 7481385 TI - The concept and classification of Guillain-Barre syndrome and related disorders. PMID- 7481386 TI - [Drug-resistant epilepsy in adults]. AB - Various estimation of the proportion of difficult-to-treat epileptic patients is given in the literature. It varies from 5 percent to 25 percent, because of imprecise meaning of pharmacoresistancy and similar terms. There is no such thing as a unique pharmacoresistant epilepsy. Persisting seizures despite antiepileptic drug prescription does not necessarily mean refractory seizures. Seizures may be drug-resistant for a limited period only, or be intermittent, or be a long lasting condition. Seizure frequency may remain unchanged, or decrease, more or less. Toxicity of AEDs must be considered. Unbearable side effects are a limiting factor for maintaining some active drugs. In practice, the term pharmacoresistancy may be used when seizures occur in spite of a convenient therapy, whatever their frequency, and whatever the resulting handicap. Predictive factors for intractibility may be considered under four headings, i.e. seizure types, patient, epileptic syndrome, and antiepileptic drugs. Most of the elements of the first three groups are available immediately after the first seizure and response to treatment may be estimated one or two years later. Outcome of an epilepsy may be evaluated, or guessed, rather early. This is crucial for management of epileptic patients. PMID- 7481378 TI - [Value of gadolinium magnetic resonance imaging of the lumbosacral roots in acute polyradiculoneuritis]. AB - Four patients with acute inflammatory polyradiculoneuropathy were evaluated with MRI. In 3 of 4 cases, gadolinium enhancement was observed in the nerve roots of cauda equina, on frontal and horizontal slices. This enhancement was correlated with the severity of the clinical picture and the cerebrospinal-fluid inflammatory protein concentration and supports the inflammatory nature of this forms of acute polyradiculoneuropathy. PMID- 7481389 TI - [Vascular dementia: clinical and radiological characteristics in 26 cases]. AB - Twenty six patients with vascular dementia were studied. Five of them only, were referred for a cognitive decline and all the others because of focal neurological symptoms or a stroke. Twelve patients had a previous history of stroke and 21 an history of high blood pressure. Twenty three patients had gait disturbances. CT scanner and MRI showed a large proportion (23 patients) of lacunar or small sub cortical strokes. Leuko-araiosis was present in 21 patients and cortical stroke only in 6. Finally, the lack of information concerning the onset and the course of the cognitive decline in 12 patients explain the difficulties to fulfill the criteria in some diagnostic evaluation scores particularly those of the DSM III R. PMID- 7481387 TI - [From the Revue Neurologique to the Societe de Neurologie: 1893-1899]. PMID- 7481388 TI - [Myoclonus of focal action and localized hemispheric lesion. A polygraphic and pharmacological study]. AB - Authors report a case of focal action myoclonus due to a localized hemispheric lesion. The woman had cardiac arrest complicated by ischemic stroke involving the left parietal region. Action myoclonus of the inferior right limb was the sole residual complaint, provoked by movement and maintenance of posture. Patient noted that myoclonus was worsened by carbamazepine and vigabatrin. EEG-EMG polygraphic study demonstrated action myoclonus and showed a particular EEG activity consisting in a volley of spikes on the vertex region during the right inferior limb movements. Contralateral movements did not elicit this activity. Pharmacological response to clonazepam, piracetam and alcohol was studied. A marked improvement of myoclonus was obtained with alcohol. Pathophysiology of this exceptional form of cortical action myoclonus is discussed. PMID- 7481390 TI - [Lhermitte-Duclos disease: is MRI diagnosis feasible?]. PMID- 7481392 TI - [Monocular circumduction nystagmus. Are the central oculomotor pathways partly monocular?]. AB - Circumduction-related nystagmus is an extremely rare and complex oculomotor anomaly. A case observed in a patient with multiple sclerosis was monocular. The lack of any motor paralysis sign demonstrated that central oculomotor pathway dysfunction can affect only one eye. This would only be possible if the central oculomotor pathways were at least in part monocular. PMID- 7481393 TI - [Pure cerebellar syndrome associated with anti-Hu antibodies]. AB - Anti-Yo is the most frequent antibody associated with pure paraneoplastic cerebellar degeneration. Cerebellar degeneration may be associated with anti-Hu antibody but other signs of nervous system involvement occur during the course of the disease. We report the case of a 69-year old man presenting with a severe acute cerebellar dysfunction of sudden onset. High titers of anti-Hu antibodies were detected in serum and cerebrospinal fluid by immunohistochemistry and western blot. No other neurological symptom was found after a follow-up of 18 months. Biopsy of a mediastinal lymph node revealed a poorly differentiated carcinoma. We suggest that the course of the anti-Hu syndrome may rarely be similar to the anti-Yo syndrome. PMID- 7481394 TI - [Unilateral benign episodic mydriasis]. AB - Ophthalmoplegic migraine is a rare phenomenon. In this setting, extrinsic as well as intrinsic fibers of the third nerve are involved. We report 2 cases of isolated, unilateral, remitting, mydriasis and describe 23 other cases already published. CT scan, MRI and angiogram were always normal when performed. The pathophysiology of this syndrome is probably related to a migrainous manifestation. PMID- 7481391 TI - [Isolated peripheral facial paralysis and HIV infection: 7 cases]. AB - Human immunodeficiency virus infection frequently involves the peripheral or central nervous system. Seven cases of isolated peripheral facial palsy that occurred at various stages of the disease are reported. The palsy was the first manifestation of the disease in 5 patients, and revealed it in 3 of them. The palsy involved the right side in 6 cases (complete N = 2; incomplete N = 4); it was bilateral in one (complete on both sides). Abnormalities of the cerebrospinal fluid were present in all patients but differed depending on the stage of the retroviral infection. Electroneuromyography defined the type of the palsy, principally due to axonotmesis in 4 patients (including the bilateral one) and principally to neurapraxia in 3 patients. The outcome was good in most cases. The physiopathology remains undetermined. PMID- 7481395 TI - [L-dopa-responsive dystonia: 2 familial cases of adult onset with sleep disorders]. AB - A woman and her son had progressive dystonia and chronic insomnia at 32 and 19 years of age respectively. Levodopa was markedly effective at low dose both for dystonia and insomnia without dyskinesia over a 5-year follow-up period. PMID- 7481397 TI - [Eosinophil meningoradiculitis caused by Angiostrongylus cantonensis]. AB - The authors report the case of a 46-year-old woman who presented with an eosinophilic meningoradiculits after a trip to Tahiti. Because of the type of presentation of the symptomatology, the CSF findings, and the spontaneously favourable outcome, it was related to an infection by Angiostrongylus cantonensis. The authors review the neurological manifestations of this parasite and emphasize the different clinical aspects of infection by Gnathostoma spinigerum, which causes myeloencephalitis in the South-East of Asia. PMID- 7481396 TI - [Transverse myelitis caused by Mycoplasma pneumoniae infection]. AB - A twenty-six year old man was admitted for febrile atypical pneumoniae. Few hours later, he presented an acute flaccid paraplegia with dorsal pain. Cerebrospinal fluid analysis showed a high leukocyte count with raised protein level. Neuroradiological examinations (myelography and MRI) were normal. Seric immunological disorders were reported. High complement-fixing antibody titers to Mycoplasma pneumoniae were found in the serum and in the CSF. The patient was treated with antibiotic and corticosteroids. He improved dramatically within one month. Neurological complications of Mycoplasma pneumoniae infections have been described (meningoencephalitis, meningitidis, polyradiculoneuropathies, cerebellar ataxia, cranial nerve palsies). Nineteen cases of transverse myelitis induced by Mycoplasma pneumoniae have been previously reported. Pathophysiological mechanisms of nervous system complications induced by Mycoplasma pneumoniae were discussed. Vascular mechanisms, direct invasion by the pathogen, toxic, immunological causes were examined. PMID- 7481399 TI - [Should asymptomatic carotid stenosis be operated on?]. AB - Should asymptomatic carotid stenosis be operated on? The aim of the present editorial is to stress that there is no simple answer to this question. The only undisputed point is that mild non hemodynamic stenosis carry such a low risk of ipsilateral brain infarct that surgery is unjustified. For tight asymptomatic stenosis ( > or = 75% at angiography according to the European method of measurement, > or = 60% according to the American method), the indication of surgery should be discussed on a case by case basis. Four main factors should be taken into account: the spontaneous risk of ipsilateral brain infarct (2% per year for stenosis > or = 75%) the risk of surgery (which should be less than 3%); the patient's life expectancy, and his own will as regards a surgical operation. It is thus possible to evaluate for each patient, the pertinence of preventive surgery and the benefit/risk ratio of the operation in order to optimize the surgical decision. PMID- 7481398 TI - [Caudate nucleus infarction]. PMID- 7481400 TI - [Conduction blocks and peripheral neuropathies]. AB - The conduction block (CB) of the nerve impulses in the peripheral nervous system induces a partial or complete loss of the nerve function, usually more marked on the motor fibers. The CB is due to a focal loss of the electrical properties of the nerve fibers. The underlying mechanisms include paranodal and segmental demyelination, and physical or chemical dysfunction of the ionic channels. The electrophysiological study allows distinction of axonal lesion from dysfunction due to CB. Recognition and quantification of the CB requires criteria that are not yet widely accepted. The different modalities of the examination are detailed and the criteria are discussed. The peripheral neuropathies with CB have different aetiologies; physical (electrical, thermal, percussion, compression or constriction injuries, radiation-induced), toxic (acting either on the axon or the myelin), more rarely ischaemic (arteriopathy or vasculitis), but frequently inflammatory (acute or chronic, primary or secondary) or related to dysimmune states (multifocal sensory-motor and motor neuropathies with persistent CB). Their review allows a discussion of their differential diagnosis in relation to the underlying mechanisms, site and duration (transient to persistent) of the CB. PMID- 7481401 TI - [Athymhormic syndrome caused by bilateral striato-capsular infarction. Moyamoya disease in adults]. AB - A 49 year-old women with no medical history suddenly presented bilateral striato capsular infarct causing frontal-like behavioural disturbances associating inertia with loss of drive, interest and affect, and preservation of intellectual function ("athymhormic syndrome" or "loss of psychic self-activation"). Ischaemic lesions mainly affected right globus pallidus and left lentiform nucleus with no involvement of the caput of the nuclei caudati. Such changes were close to anoxic lesions known to cause the same symptomatology. Infarction was attributed to a Moyamoya disease on angiographic data. Bilateral involvement of basal ganglia, arterial borderzones or both, may explain the frequent occurrence of neuropsychological disturbances in Moyamoya disease. PMID- 7481402 TI - [Athymhormic syndrome and progressive paraplegia disclosing adrenoleukomyeloneuropathy in an adult]. AB - A 40-year-old man presented with both paraparesia and an athymhormic syndrome. Bicapsular lesions together with abnormal evoked motor potentials suggested cortical and spinal involvement in the deficiency syndrome. The neuropsychologic symptoms appeared to be secondary to bilateral pallidium lesions. Associated signs were adrenoleukocystrophy and adrenomyeloneuropathy suggesting the term, adrenoleukocyeloneuropathy. PMID- 7481403 TI - [Kufs disease with leukoencephalopathy]. AB - We report a case of adult neuronal ceroid lipofuscinosis (Kufs' disease) with leukoencephalopathy on cerebral scan CT and MRI. A 52 year-old woman presented with partial complex epileptic seizure followed by progressive dementia, cerebellar ataxia, pyramidal and akineto-rigid signs and symptoms. After 6 years of evolution, cerebral stereotactic biopsies showed a diffuse gliosis of the white matter, but no clear demyelination. Nerve and glial cells contained numerous PAS+ autofluorescent granules. In the oligodendrocytes and astrocytes of the white matter these granules appeared electronmicroscopically as cytoplasmic osmiophilic lamellar bodies with fingerprint profile combined with some curvilinear and rectilinear aspects. The cortical nerve cells contained granular osmiophilic bodies. This "leukoencephalopathic" variant of Kufs' disease is probably related to the pigmentary type of orthochromatic leukodystrophy, wherein similar inclusions have been only described in the macrophages and glial cells of the white matter. PMID- 7481404 TI - [Multiple system atrophy with Lewy bodies]. AB - The term multiple system atrophy has been used to define a unique sporadic neurodegenerative disease, usually occurring in midlife, pathologically characterized by degeneration of the nigro-striato-pallidal and olivo-ponto cerebellar systems and autonomic neurons of the spinal cord, and by the presence of characteristic oligodendroglial inclusions. In many cases, this disease can be readily distinguished, both clinically and pathologically, from idiopathic Parkinson's disease. However, often clinical differentiation is difficult, and in a few autopsied cases the presence of Lewy bodies, the characteristic inclusion of idiopathic Parkinson's disease, can lead to diagnostic confusion. Such a pathological association, named the "transitional variant" by some authors, is of unknown clinical and pathological significance. We here report three new cases of such an association from the United Kingdom Parkinson's Disease Society Brain Bank. Clinical and pathological data derived from these new observations, as well as from literature cases, tend to suggest that this pathological association is of no particular clinical significance. The comparison of the prevalence of Lewy bodies in normal elderly individuals and in multiple system atrophy suggests a chance association of the two pathologies. However, the small number of cases so far studied as well as the heterogeneity of series analyzed does not exclude the presence of common susceptibility factors for both diseases. PMID- 7481405 TI - [N-acetyl-aspartate abnormalities in internal-temporal epileptic foci using proton magnetic resonance spectroscopy]. AB - The aim of this study was to characterize the neurochemical abnormalities related to N-acetyl-aspartate which is a neuronal marker, within an epilepticus focus located in the internal-temporal area, using proton magnetic resonance spectroscopy. Eleven patients with a mono-hippocampal epileptics focus on clinical and per-critical electroencephalographical criteria, were matched with 11 controls by age, sex and laterality. Proton spectroscopy of a volume of 8 cm3 was performed within the ipsilateral and the contralateral internal-temporal area and within the 2 hippocampus of controls. Volumetry of the ipsilateral and the contralateral hippocampus and of the 2 hippocampus of controls was performed using resonance magnetic imaging. All these measurements were performed during the interictal stage. The results were concordant to show a decrease of the ratio N-acetyl-aspartate/choline and N-acetyl-aspartate/creatine within the epilepticus focus, in relation with a hippocampal atrophy. This study finds similar results to those of other previous works. The decrease of N-acetyl-aspartate levels within the epilepticus focus could be related to a decrease of the neuronal cell density. This procedure is able to show a decrease of the levels of this metabolite within an internal temporal epilepticus focus and associated with a hippocampal atrophy. PMID- 7481406 TI - [Cognition disorders and parkinsonian syndrome: diffuse Lewy body disease?]. AB - A 68 year old man developed progressive cognitive impairment with an akinetic rigid syndrome which was atypically responsive to levodopa. The patient died after 18 months and the postmortem examination showed typical Lewy bodies in the substantia nigra and pale bodies in the cortex which were unlabelled by ubiquitin antibodies. This case is particular by the neuropathological examination (no immunoreactivity to antibodies against ubiquitin) and by the abnormal response to treatment (dystonia). These two aspects are discussed. PMID- 7481407 TI - [Dacrystic and asystolic epileptic seizures]. AB - A 33-year-man with an encephalopathy of unknown aetiology, had an history of epilepsia for 30 years. Different types of seizures were seen, including grand mal and frontal attacks. Epilepsia was associated with mental retardation and behavioral disorders. At the age of 33, he was admitted for repetitive general convulsions. Epilepticus status lasted for two weeks and improved with vigabatrin et clonazepam. General seizures, frontal motor convulsions with arms and trunk antepulsion, and dacrystic attacks were seen. The latter seemed to be like normal crying because they were accompanied by lacrimation, contorted and mournful facies, and sobbing sounds. One year later, repetitive cardiac arrests occurred during a new epilepticus status. Cardiac arrests, observed on ECG holter lasted 10 to 24 seconds, without cardiac dysfunction. EEG patterns on ECG holter lasted 10 to 24 seconds, without cardiac dysfunction. EEG patterns included theta and delta activity with rhythmic slow wave epileptic activity, predominating on right side, in temporal areas. CT scan was normal. MRI showed right cerebral atrophy, prevailing in the temporo-mesial region, with right temporal horn enlargement. This case report of dacrystic seizures, the first one with MRI study, suggests that temporo-mesial structures of the non-dominant hemisphere may be involved in dacrystic and asystolic attacks. PMID- 7481408 TI - [Hemiballismus with logorrhea and thymo-affective disinhibition caused by hematoma of the left subthalamic nucleus]. AB - A 71 year-old hypertensive right-handed man presented a small intracerebral haematoma limited to the left subthalamic nucleus that resulted in right hemiballismus and a neurobehavioral disinhibition syndrome. The latter associated logorrhoea and euphoria, but there was no frontal signs nor mental confusion. The patient kept full insight into his mental change, that contrasted with his prior personality. Hemiballismus and the manic-like state resolved within 2 months. This case and some others from the literature illustrated that a focal lesion in the basal ganglia could result in manic-like affective change, and that a relation did exist between hyperkinetic syndrome and euphoria. Our case contrasted with most of the published cases of neurobevioral changes following basal ganglia discrete lesions, expressing as hypokinesia, apathy and loss of motivation. With reference to the functional circuits linking the frontal cortex and the basal ganglia, as described by Alexander, this syndrome of combined disinhibition of motricity, speech and affect could involve either the lateral orbito-frontal or the anterior cingulate loops, more especially the indirect pathways travelling through the subthalamic nucleus, lesion of which is known to disinhibit thalamo-cortical activity. PMID- 7481409 TI - [Conference at the Salpetriere. April 1994 . Familial dementia and onirism in a 63-year-old woman]. PMID- 7481410 TI - Circadian rhythm adaptation to simulated night shift work: effect of nocturnal bright-light duration. AB - We compared bright-light durations of 6, 3 and 0 hours (i.e. dim light) during simulated night shifts for phase shifting the circadian rectal temperature rhythm to align with a 12-hour shift of the sleep schedule. After 10 baseline days there were 8 consecutive night-work, day-sleep days, with 8-hour sleep (dark) periods. The bright light (about 5,000 lux, around the baseline temperature minimum) was used during all 8 night shifts, and dim light was < 500 lux. This was a field study in which subjects (n = 46) went outside after the night shifts and slept at home. Substantial circadian adaptation (i.e. a large cumulative temperature rhythm phase shift) was produced in many subjects in the bright light groups, but not in the dim light group. Six and 3 hours of bright light were each significantly better than dim light for phase shifting the temperature rhythm, but there was no significant difference between 6 and 3 hours. Thus, durations > 3 hours are probably not necessary in similar shift-work situations. Larger temperature rhythm phase shifts were associated with better subjective daytime sleep, less subjective fatigue and better overall mood. PMID- 7481411 TI - Effects of timed bright-light exposure on shift-work adaptation in middle-aged subjects. AB - Shift workers suffer from a constellation of symptoms that can severely compromise their ability to perform optimally on-shift. The largest single factor contributing to shift-worker problems is sleep disturbance, and there is little question that the primary cause of such sleep disturbance is circadian disruption. Recently, a number of studies have demonstrated that timed exposure to bright light can help facilitate adaptation to simulated shift-work schedules, at least in younger subjects. The aim of the current study was to assess the effects of bright-light interventions in middle-aged individuals undergoing a simulated shift-work regimen. Results indicate that although light was effective in resetting the circadian clocks of these subjects by more than 6 hours, there was little effect on measures of on-duty alertness and performance or on off-duty sleep. These findings suggest that middle-aged subjects may be less phase tolerant than young subjects, and they raise questions concerning the utility of bright-light interventions in some shift-work populations. PMID- 7481412 TI - Effects of bedtime administration of zolpidem on circadian and sleep-related hormonal profiles in normal women. AB - Short-acting benzodiazepine hypnotics may phase-shift circadian rhythms and improve adaptation of sleep patterns to abrupt time shifts, depending on the timing of administration. The aim of the present study was to determine whether bedtime administration of zolpidem, a non-benzodiazepine hypnotic, causes alterations in circadian rhythmicity or in the normal interactions between sleep and hormones. Eight normal women (aged 21-33 years) each participated in a baseline study and a study with zolpidem administration. On each occasion, blood samples were obtained at 20-minute intervals for 25 hours, starting at 1000 hours. Zolpidem (10 mg) was given orally at 2245 hours. Zolpidem administration was associated with an increase in stages III + IV sleep. Cortisol, melatonin, thyrotropin and growth hormone profiles were similar in both experimental conditions. In contrast, though remaining in the normal range, the nocturnal elevation of prolactin was enhanced two-fold in all subjects after zolpidem during early sleep, and prolactin levels were still 50% higher than baseline in late sleep. Morning levels were similar in both studies. In conclusion, bedtime administration of 10 mg zolpidem, a standard clinical dosage, systematically induces a transient moderate hyperprolactinemia, but does not alter other sleep related hormonal secretions or endocrine markers of circadian rhythmicity. PMID- 7481413 TI - Sleep complaints among elderly persons: an epidemiologic study of three communities. AB - The frequencies of five common sleep complaints--trouble falling asleep, waking up, awaking too early, needing to nap and not feeling rested--were assessed in over 9,000 participants aged 65 years and older in the National Institute on Aging's multicentered study entitled "Established Populations for Epidemiologic Studies of the Elderly" (EPESE). Less than 20% of the participants in each community rarely or never had any complaints, whereas over half reported at least one of these complaints as occurring most of the time. Between 23% and 34% had symptoms of insomnia, and between 7% and 15% percent rarely or never felt rested after waking up in the morning. In multivariate analyses, sleep complaints were associated with an increasing number of respiratory symptoms, physical disabilities, nonprescription medications, depressive symptoms and poorer self perceived health. Sleep disturbances, particularly among older persons, oftentimes may be secondary to coexisting diseases. Determining the prevalence of specific sleep disorders, independent of health status, will require the development of more sophisticated and objective measures of sleep disturbances. PMID- 7481414 TI - Four-year follow-up of mortality and sleep-related respiratory disturbance in non demented seniors. AB - To ascertain whether sleep-disordered breathing (SDB) in the elderly is associated with increased mortality, a prospective cohort study with 4-year follow-up was conducted at a retirement village complex in Sydney, Australia. The subjects were 163 non-demented retirement village residents. Logistic regression was used to assess SDB and co-morbidity as independent predictors of mortality. Respiratory disturbance index (RDI) was measured in the home; those subjects with RDIs > or = 15 were classified as having SDB. Co-morbidity was measured by an index of Burden of Illness based on the medical history obtained at baseline. At 4 years, 27% (4/15) of those subjects with RDIs > or = 15 and 22% (33/148) of those with RDIs < 15 were dead. RDI had an odds ratio (OR) of 1.00 (95% CI: 0.96, 1.04). Burden of Illness had an OR of 1.90 (95% CI: 1.34, 2.71). Adjustment for age and sex did not alter these findings. Significant predictors of mortality from the illness measure were a history of hypertension, Parkinson's disease and other severe illnesses (usually cancer). RDI was not a predictor of mortality in this population of non-demented seniors, where the prevalence of high levels of RDI was low. PMID- 7481415 TI - Automated scoring of sleep in the neonatal lamb. AB - The study of sleep is an important and rapidly expanding area of research that generates large bodies of data. Manual scoring of sleep states from polygraph recordings is a laborious and often subjective task. Even when care is taken, the opportunity for disagreement between investigators and between laboratories remains great. To avoid this difficulty and to reduce the subjectivity of sleep state scoring we have designed a computer-based algorithm for scoring sleep state in the lamb. The algorithm underlying the system relies upon spectral analysis of the electrocorticogram and upon amplitude analysis of the electrooculogram and nuchal electromyogram. Partitioning the spectral power observed within the electrocorticogram (1-4 Hz frequency range) reliably identifies deep quiet sleep. Wakefulness and active sleep are then identified based upon threshold crossings of the electrooculogram and of the electromyogram of the nuchal muscles. We compared the sleep states returned by the algorithm to those scored visually by trained personnel for 1 hour of data collected from each of five 19-day-old lambs. There was good agreement between the two methods of scoring sleep. The percents agreement between the algorithm-derived scores and visual scores were as follows: active wakefulness 97%, quiet wakefulness 87%, quite sleep 85% and phasic active sleep 82%. As such, our algorithm provides a fast, reliable and objective method for scoring sleep state in the young lamb. PMID- 7481416 TI - Slow wave sleep and cyclic alternating pattern (CAP) in HIV-infected asymptomatic men. AB - Alterations of sleep structure have been reported in asymptomatic human immunodeficiency virus (HIV)-infected subjects. In these patients some authors have found an increased percentage of slow wave sleep (SWS) and a SWS preponderance in the second half of the night, as well as subjective sleep complaints. Other authors have found an increased stage 1 non-rapid eye movement (NREM) and reduced stage 2 NREM percentages in asymptomatic subjects. We evaluated the macrostructure and the microstructure (cyclic alternating pattern, CAP) of sleep in nine HIV-infected asymptomatic men without sleep complaints or psychiatric illness, in comparison with nine age-matched controls. Our study showed a decreased amount of SWS and a significantly higher CAP rate in HIV subjects, suggesting an altered organization of the sleep process in these patients. PMID- 7481417 TI - Sleep/waking and EEG power spectrum effects of a nonselective serotonin (5-HT) antagonist and a selective 5-HT reuptake inhibitor given alone and in combination. AB - Sleep/waking stages, electroencephalogram (EEG) power spectra and behavior were studied in rats for 8 hours following intraperitoneal administration of a nonselective serotonin (5-HT) antagonist (0.1 and 2.0 mg/kg methiothepin) and a selective 5-HT reuptake inhibitor (20 mg/kg zimeldine), given alone and in combination. Consistent with earlier studies, zimeldine gave a biphasic effect on sleep and waking. Waking was increased and slow wave sleep (SWS)-2 decreased initially, followed by an increase in SWS-2 in the second 2-hour period. Rapid eye movement (REM) sleep was reduced throughout the experiment. EEG power densities were generally reduced in the higher frequencies, but the effect differed somewhat in the different vigilance states and between the fronto frontal and fronto-parietal EEG leads. Zimeldine did not change behavior. Methiothepin, at 0.1 mg/kg, gave only minor effects by itself, but it blocked the initial waking increase of zimeldine. So did 2.0 mg/kg methiothepin, but this dose markedly changed sleep/waking stages by itself: SWS-1 was profoundly increased, whereas waking, SWS-2 and REM sleep were reduced. Total SWS (TSWS) was markedly increased due to the SWS-1 increase. Because TSWS was increased while SWS-2 was decreased following 2.0 mg/kg methiothepin, it is concluded that spindle activity was facilitated, whereas slow wave activity was antagonized. Methiothepin, at 2.0 mg/kg, also markedly changed EEG power densities within TSWS and induced cataleptic behavior. It is concluded that the initial waking increase of zimeldine depends on simultaneous activation of several different 5-HT receptor subtypes. The other zimeldine effects were not consistently antagonized, thus the mechanisms behind these effects remain unclear. PMID- 7481418 TI - First-night effect in normal subjects and psychiatric inpatients. AB - The goal of the present study was to evaluate the first-night effect in psychiatric inpatients using large subject samples (n > 30) in order to obtain a good statistical evaluation. Thirty-two normal subjects and 94 psychiatric inpatients (38 depressives and 56 insomniacs) were studied for three consecutive nights in the hospital sleep laboratory. Our results showed clearly that there was a first-night effect in normal subjects, similar to that reported in previously published data, characterized by a longer rapid eye movement (REM) sleep latency (p < 0.05), increased wakefulness (p < 0.01) and total sleep time (p < 0.02) and a decreased sleep efficiency (p < 0.01). REM sleep latency and stage REM in the first third of the night were still altered in the second night. Both clinical groups had a less marked first-night effect than normal subjects, showing alterations only observed in REM sleep (p < 0.01) (decreased REM sleep, longer REM sleep latency, increased REM sleep gravity center). However, the first night effect was more pronounced in insomniacs than in depressed patients. No statistical differences between the second and third nights' recordings were found in sleep parameters. It is suggested that first-night data should not be simply discarded but could be used in subsequent analyses. PMID- 7481419 TI - Acute, subchronic and withdrawal sleep EEG changes during treatment with paroxetine and amitriptyline: a double-blind randomized trial in major depression. AB - Paroxetine (30 mg), a selective serotonin (5-HT) reuptake inhibitor, was compared in a double-blind trial to amitriptyline (150 mg) in a sample of 40 inpatients aged 18-65 years who fulfilled Research Diagnostic Criteria for major depression. Patients were studied after a placebo drug washout period of 10 days and after an active 4-week treatment period. Sleep EEG recordings were performed before and at the end of the study as well as during acute treatment (first 2 days) and following withdrawal of active medication. Paroxetine shows an antidepressant effect similar to amitriptyline with a different side-effect profile typical of 5 HT reuptake inhibition. Paroxetine and amitriptyline decreased the amount of REM sleep, a well-known effect of classical antidepressants. Paroxetine also shared with other 5-HT reuptake inhibitors an alerting effect on sleep that was not shown to be detrimental on subjective sleep quality. PMID- 7481420 TI - Magnocellular nuclei of the basal forebrain: substrates of sleep and arousal regulation. AB - Magnocellular regions of the basal forebrain (BF) are recognized as important sites of sleep-wake regulation. Evidence is reviewed for the coexistence within the BF of mechanisms that regulate neocortical and limbic system arousal along with mechanisms that promote sleep. Arousal-related functions are mediated by a system of magnocellular cholinergic neurons. BF cholinergic neurons project monosynaptically to the entire neocortex and limbic telencephalon, exert excitatory effects on target cells and participate in the regulation of activated EEG patterns characteristic of waking and REM sleep. Evidence suggests that, within the waking state, the BF cholinergic system modulates processing of sensory information in the neocortex and is involved in cognitive processes. One or more noncholinergic cell types are responsible for the sleep-promoting functions of the BF. Neurons that display elevated discharge rates during transitions from waking to sleep and during nonREM sleep have been recorded in BF sites were electrical stimulation evokes sleep and experimental lesions cause insomnia. BF neurons function to promote sleep, in part, via descending inhibition of caudal hypothalamic and brainstem activating systems. GABAergic neurons located within magnocellular regions of the BF are hypothesized to mediate sleep-promoting actions. Afferents to the BF from hypothalamic and brainstem regions are functionally important for sleep-wake regulation. Thermosensitive inputs from the anterior hypothalamus modulate the activity of BF sleep- and arousal-related cell types. Excitatory effects of brainstem inputs to BF arousal-related cells have been documented. Additional evidence supports a critical role for GABAergic-cholinergic interactions, both within the magnocellular BF and at cortical and diencephalic sites, in the regulation of behavioral state. PMID- 7481421 TI - Oral appliances for the treatment of snoring and obstructive sleep apnea: a review. AB - This paper, which has been reviewed and approved by the Board of Directors of the American Sleep Disorders Association, provides the background for the Standards of Practice Committee's parameters for the practice of sleep medicine in North America. The 21 publications selected for this review describe 320 patients treated with oral appliances for snoring and obstructive sleep apnea. The appliances modify the upper airway by changing the posture of the mandible and tongue. Despite considerable variation in the design of these appliances, the clinical effects are remarkably consistent. Snoring is improved and often eliminated in almost all patients who use oral appliances. Obstructive sleep apnea improves in the majority of patients; the mean apnea-hypopnea index (AHI) in this group of patients was reduced from 47 to 19. Approximately half of treated patients achieved an AHI of < 10; however, as many as 40% of those treated were left with significantly elevated AHIs. Improvement in sleep quality and sleepiness reflects the effect on breathing. Limited follow-up data indicate that oral discomfort is a common but tolerable side effect, that dental and mandibular complications appear to be uncommon and that long-term compliance varies from 50% to 100% of patients. Comparison of the risk and benefit of oral appliance therapy with the other available treatments suggests that oral appliances present a useful alternative to continuous positive airway pressure (CPAP), especially for patients with simple snoring and patients with obstructive sleep apnea who cannot tolerate CPAP therapy. PMID- 7481424 TI - Bibliography of recent literature in sleep research. PMID- 7481422 TI - Practice parameters for the treatment of snoring and obstructive sleep apnea with oral appliances. American Sleep Disorders Association. AB - These clinical guidelines, which have been reviewed and approved by the Board of Directors of the American Sleep Disorders Association (ASDA), provide recommendations for the practice of sleep medicine in North American with regards to the use of oral appliances for the treatment of snoring and obstructive sleep apnea. Oral appliances have been developed for the treatment of snoring and have been applied to the treatment of obstructive sleep apnea, a syndrome associated with morbidity. Based on a review of the relevant scientific literature, the Standards of Practice Committee of the ASDA has developed guidelines describing the use of oral appliances for the treatment of snoring and obstructive sleep apnea in adults. PMID- 7481423 TI - Sleep and breathing in amyotrophic lateral sclerosis. PMID- 7481425 TI - [Ethical guidelines for human research sponsored by FAPESP. Group of Bioethics and Populational Research Fundacao de Amparo a Pesquisa do Estado de Sao Paulo]. PMID- 7481426 TI - Treatment of transfixing hepatic lesions with a hydrostatic balloon. PMID- 7481427 TI - E test: a novel technique for antimicrobial susceptibility testing. AB - We describe the applicability of the E test (AB Biodisk Solna, Sweden), a new method for determining minimum inhibitory concentrations of antimicrobial agents against bacteria. This report is based on the literature review and on our own experience using the E test for susceptibility testing of the Xanthomonas maltophilia, Streptococcus pneumoniae and Streptococcus viridans group against eight different drugs. PMID- 7481428 TI - Aspergillary bronchopneumonia: an unusual cause of atelectasis and asphyxia in a leukemic patient. AB - A 22-year-old man in his first relapse of T-acute lymphoblastic leukemia developed fever and a pulmonary infiltrate after 23 days of granulocytopenia. Although having been under amphotericin B for 10 days, productive purulent cough ensued, with right lobe atelectasis and acute ventilatory failure that resolved after the elimination of a thick gelatinous bronchial plug. Sputum cultures yielded Candida Albicans and Staphylococcus epidermidis, and microscopic examination of the sputum plug disclosed Aspergillus hyphae. The patient died 9 days after, of a disseminated Aspergillus infection, confirmed by necropsy. PMID- 7481429 TI - Nucleolar organizer regions (NORs) AB - Counts of nucleolar regions (NORs) demonstrated by a silver staining technique in paraffin sections, have been used to distinguish between benign and malignant lesions. AgNORs were studied in 24 biopsies from oral cavity (5 cases of normal oral mucosa, 5 of pseudocarcinomatous hyperplasia and 14 of squamous cell carcinoma, subdivided according to degree of differentiation: 5 grade 1, 5 grade 2 and 4 grade 3) to find whether they were helpful in distinguishing pseudocarcinomatous hyperplasia due to chronic parasitic infections from squamous cell carcinoma. Two methods of counting AgNORs were used: (A) a simpler one which counts nucleolar clusters (AgNU) and satellite AgNORs and the other (B) counting all individual AgNORs, including those within AgNUs. In both methods the lowest mean values were observed for grade 3 carcinoma, while the highest belonged to grade 2 carcinoma. The simpler method (A) was the most useful because AgNU counts showed significant difference when pseudocarcinomatous hyperplasia was compared with grade 1 and 2 carcinomas, which are the most difficult to discriminate from it. However the overlapping of values render the technique of limited use in individual cases. PMID- 7481430 TI - Neurological complications of coronary arteriography. AB - The authors report a 46 year-old female who was submitted to a right femoral artery coronary arteriography. During the procedure she presented an upward, downward and convergence gaze paresis; an upbeating nystagmus. A NMRI showed a change in T-2 weighted signal of the left paramedian mesencephalic region. A diagnosis of stroke was made and dextran 40 was prescribed. Vertical gaze paresis progressively disappeared. Convergence paresis and nystagmus improved partially only. The possible pathophysiological basis underlying this clinical picture are: catheter-induced dislodgement of an atheromatous plaque; small thrombi formation and release from the catheter tip; catheter-induced vascular lesion with subsequent thromboembolism or dissection of the intima; cardiac arrhythmia; catheter-induced or contrast medium-induced arterial spasm; a combination of two or more of these. PMID- 7481431 TI - Fabry's disease in a female carrier with bilateral thalamic infarcts: a case report and a family study. AB - An unusual case of a young woman, heterozygote for Fabry gene is reported, who presented bilateral thalamic infarcts due to occlusions of central nervous system vessels. Three other members of her family were studied. Fabry's disease (angiokeratoma corporis diffusum) is included among the rare causes of ischemic stroke in young adults. PMID- 7481432 TI - Intercoronary connection with bidirectional blood flow and concentric left ventricular hypertrophy. AB - Cardiac catheterization in a 55-year-old man, with a 6-month history of atypical chest pain and Q-waves in DII, DIII and AVF, showed concentric left ventricular (LV) hypertrophy and a large intercoronary connection between right coronary artery (RCA) and circumflex artery (CX), with bidirectional blood flow. Although the RCA and CX were normal, selective injection of CX filled RCA retrogradely and in the same way selective injection of RCA filled CX. Possible mechanisms and literature are reviewed. PMID- 7481434 TI - Limb salvage procedures in osteosarcomas around the knee joint. AB - Surgical techniques developed for restoring skeletal continuity after a local resection of a bone tumor (limb salvage procedures) revolutionized the management of the patient suffering from osteosarcomas around the knee. In this article the authors review the current status of various reconstruction procedures; they include osteoarticular allograft arthrodesis, rotation plasty, expandable prosthesis and knee arthroplasty. In summary they suggest that the knee arthrodesis is the preferred procedure for young and active patients, the rotation plasty should be recommended as the alternative to amputation for very young patients, and the customized prostheses are preferred for patients with limited longevity. PMID- 7481435 TI - Facial bone lengthening apparatus with expander and miniplates. AB - Since 1869, bone lengthening has been pursued; when Langenbeck tried growing a shortened bone extremity utilizing for this an arteriovenous fistula in the thigh. At the beginning of this century, codovilla (1905) introduced bone lengthening methods for lower limbs. PMID- 7481433 TI - Restoration of sensitivity after removal of the sural nerve. A new application of latero-terminal neurorraphy. PMID- 7481436 TI - [Laser in gynecology. Palliative laser treatment in genital and breast cancers]. AB - Local, so-called incurable, recurrences of breast or genital cancers were as therapeutic challenge in the past. The authors undertook a pilot trial of laser treatment in 45 patients, as a palliative measure, in view of the serious psychological and physical problems experienced by these women constantly confronted by the presence of painful, visible and often ulcerated tumor lesions. CO2 and Nd:YAG lasers were used for tumor vaporisation and coagulation, the combined use of these two wavelengths providing the new study concept which emerged from this pilot trial. PMID- 7481437 TI - [Ovarian cancer and pregnancy]. AB - The authors report 6 cases of carcinoma of the ovary presenting during pregnancy in the Department of Gynecology and Obstetrics of La Grave University Hospital, Toulouse, between 1972 and 1994. Analysis of this series is followed by a review of the literature which reveals that carcinoma of the ovary fortunately remains a rare event during pregnancy. All reported series conclude that the diagnosis is made at an early stage. Management depends in great part on the age of the pregnancy and stage of malignancy. During the first 6 months, immediate surgery is essential, with sacrifice of the pregnancy, except at stage IA1. During the final 3 months, surgery is delayed until fetal maturity. This classical attitude is currently being questioned. This classical attitude is currently being questioned. Some teams feel that conservative treatment followed by chemotherapy is possible, thereby preserving the pregnancy. The choice of the couple must always be taken into account when making these management decisions. PMID- 7481438 TI - [Preliminary clinical study on the management of 26-34 week pregnancy with premature rupture of the membranes (PRM)]. AB - The aim of this study was to determine the optimal solution to the dilemma of "wait-and-see" or "evacuation of uterine contents" and to identify objective criteria in support of one or other of these management choices, in particular in 26-34 week pregnancies with PRM, which is the most difficult situation. On the basis of a clinical study of 54 cases, week pregnancies with PRM did not enable temporization for any considerable time because of the onset of amniotic infection, and a large number of premature newborn with a gestational age of 31 32 weeks, below 1500-1700 g respectively, died a short time after birth. CONCLUSIONS: In 26-34 week pregnancies with PRM, a "wait-and-see" attitude with its sometimes very considerable septic risks is justified by a high chance of fetal survival score (CFSS)--a concept suggested by the authors--, i.e. with a gestational age between 30-34 weeks and respective fetal weights (determined by ultrasonography) between 1400-2100 g, absence of intrauterine fetal pathology, favorable fetal vitality parameters, good prognosis foe vaginal delivery, low C reactive protein levels, low-grade pathogenicity of cervicovaginal flora, etc. In their preliminary study, the authors suggest determination of a chance of fetal survival score (CFSS) which is of prognostic value and can be useful in determining optimal management in the dilemma raised by PRM in 26-34 week pregnancies. PMID- 7481440 TI - [Cervical pregnancy. Contribution of ultrasonography to diagnosis and therapeutic management]. AB - Three recent cases of cervical pregnancy form the basis of a review of its principal clinical and ultrasonographic characteristics and their resultant management consequences. Ultrasonography enables better diagnosis and management, leading to the preservation of subsequent fertility. PMID- 7481439 TI - [A comparative study of 2 ways of clinical management in premature rupture of the membranes at term: temporization versus labor induction]. AB - Two defined management approaches, temporization limited to 48 hours and immediate induction of labor, for premature rupture of the membranes at term were compared in a prospective study between January 1 1991 and November 30 1993 in the Maternity Unit of Yaounde University Hospital. During this period, 268 cases of premature rupture of the membranes were seen among 3252 deliveries, i.e. an incidence of 8.2%. In the temporization group (153 cases), spontaneous onset of labor was effective in 95 patients (62.1%) within 12 hours and in 137 patients (89.5%) within 24 hours after premature rupture of the membranes. Spontaneous deliveries in this temporization group accounted for 129 cases (92.8%). In the induction of labor group, spontaneous delivery occurred in 119 cases (93.2%). There were ten cesareans in the temporization group and eight cesareans and two vacuum cup extractions in the induction group. Short term (24 hours) prophylactic antibiotics were given to 34 patients, i.e. 16 cases in which the duration of rupture of the membranes was more than 24 hours and 18 cases of cesarean section. Maternal infections concerned 18 cases (6.7%) including 12 cases (4.4%) of malaria. Microbiology of vaginal swabs revealed 6 cases of pseudomonas, 4 cases of staphylococcus aureus and 3 of candida albicans. Neonatal infections confirmed by blood culture and assay of C-reactive-protein involved 24 cases (20.3%). There were three fetal deaths, i.e. perinatal mortality of 1.1%. Risk factors, in these three fetal deaths, included postmaturity (1 case).(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7481441 TI - [Simultaneous intrauterine and extrauterine pregnancy in the presence of inert intrauterine device: a case report]. AB - The authors report a case of simultaneous intra-uterine and ectopic pregnancy in the presence of a Lippes loop (inert intra-uterine device) which appeared to have been well tolerated for four years. Predisposing factors appeared to be infection, evidenced by the peri-adnexal adhesions found at surgery for the ectopic pregnancy, together with spontaneous multiple ovulation. The intra uterine pregnancy culminated in a spontaneous abortion. Although apparently very rare, the association of intra-uterine pregnancy and ectopic pregnancy in the presence of an intra-uterine device should be born in mind whenever intrauterine contraception fails. PMID- 7481442 TI - [Uterine inversion: a cause of obstetric hemorrhage not to be missed. Discussion]. AB - Severe obstetric bleeding is the primary cause of maternal morbidity and mortality. While classical causes such as uterine inertia or retained placenta are well known, inversion of the uterus is a rare cause which should not be missed. Only very rapid diagnosis and management can avoid serious and irreversible consequences. A case of inversion of the uterus is reported. The circumstances of onset and treatment possibilities are also discussed. PMID- 7481443 TI - [Sacrococcygeal teratomas. Apropos of a case of teratoma weighing 4200 g]. AB - We report a clinical case of a large sacro-coccygeal teratoma diagnosed antenatally by ultra-sound at 35 weeks gestation. A review of the literature shows that their outcome depends upon the size of the tumor, its degree of maturity, whether its location is pelvic or not, the presence of any associated malformations, prematurity, atraumatic delivery and upon prompt and complete removal of the tumor. PMID- 7481444 TI - [What about partial subcutaneous symphysiotomy or Zarate's operation? Review of the literature]. AB - This study is a review of the literature on Zarate's partial subcutaneous symphysiotomy. Unfortunately, this operation is now forgotten since many obstetricians in western countries and even those of developing countries feel that symphysiotomy is an obsolete obstetric procedure. Partial subcutaneous symphysiotomy is an operation intended to widen the maternal pelvis in order to enable vaginal delivery in the presence of moderate feto-pelvic disproportion. The procedure, which requires only a urinary catheter, a scalpel and local anesthesia, consists of division of the anterior ligament of the public symphysis and part of the inferior ligament (arcuate ligament of the pelvis), while leaving intact the superior ligament or "anatomo-surgical restraint" of Zarate, ensuring limitation of pelvic spread. Symphysiotomy has currently been abandoned by all or almost all teams because of its alleged maternal and fetal morbidity and mortality. We feel, however, that in certain rural areas of third world countries, often ill-equipped and with poor medical services, this procedure can still be very useful in certain exceptional situations in dystocia by moderate feto-pelvic disproportion, in circumstances in which cesarean section is impossible. The experience of the rare teams still using symphysiotomy shows that a degree of disdain is legitimate in face of the ominous reservations expressed as to the possible complications of this procedure. PMID- 7481445 TI - [Gynecologic monitoring of tamoxifen treatment of breast cancer]. PMID- 7481446 TI - [Management of difficult cases of biliary lithiasis]. AB - To assess that insertion of a biliary endoprosthesis is a safe and effective treatment in patients with retained common bile duct stones (CBDS). BACKGROUND: Before 1974, surgical exploration including cholecystectomy with choledochotomy was the only effective treatment for CBDS. However, morbi-mortality increases rapidly in the elderly and is over 10% in patients over 65, particularly if other serious medical conditions are present. Endoscopic sphincterectomy is now an established procedure, with a success rate of about 90%, complications are relatively rare, and mortality is about 1%. However, the size and number of the stones, the appearance of the terminal bile duct and presence of peripapillary diverticulum influence the outcome. At the present time technique of fragmenting the stones using intracorporeal electrohydraulic, extracorporeal shock waves or laser lithotripsy would be seen to be an attractive alternative approach. Endoprosthesis insertion is a safe, effective and cheap treatment in CBDS. PATIENTS, METHODS AND RESULTS: 40 patients with unextractable CBDS were studied and treated with endoscopic stenting. Four patients were lost for follow-up. The remaining patients comprised 29 females and 7 males, with a mean age of 67 years. Four patients underwent surgical treatment for post-sphincterotomy haemorrhage (1 patient), internal endoprosthesis migration (1 patient) and peripapillary diverticulum (2 patients). In 7 high-risk patients with large CBDS, recurrent obstructive jaundice was successfully treated by replacing the endoprosthesis. In 6 of them we added medical treatment with ursodeoxycholic acid and after 24 months follow-up, the stones disappeared. In 13 patients the endoprosthesis were inserted temporally, and in a second approach we removed the CBDS by mechanical lithotripsy. The remaining 12 patients were still alive without symptoms after a follow-up of 24 months. CONCLUSIONS: On the basis of these findings we therefore concluded that endoscopic insertion of a biliary endoprosthesis is a safe and effective treatment for CBDS in patients in whom endoscopic sphincterotomy and attemps to remove the stones are not successful. PMID- 7481447 TI - [Complications of splenectomy in the Sugiura-Futagawa procedure]. AB - Evaluation of the complications of splenectomy in the Sugiura-Futawaga operation. BACKGROUND: Splenectomy in portal hypertension is a more difficult procedure and probably has a higher morbidity and mortality. METHOD: A retrospective study of 155 patients with hemorrhagic portal hypertension treated by means of the Sugiura Futawaga operation was conducted to evaluate the complications related to splenectomy. RESULTS: Mean age of the patients was 42 years: 81 patients were male and 74 female. In 87 cases liver cirrhosis was shown, 39 cases with portal fibrosis, 23 cases with primary biliary cirrhosis and 6 cases with normal liver (idiopathic portal hypertension). No patient received preoperative immunization against pneumococcus, but received antibiotic prophylaxis. Complications were found in 16 cases (10.3%): 13 of them related to the surgical procedure (peritonitis 3.9%, subphrenic collection 1.9%, pancreatic pseudocyst 1.9% and pancreatic fistula 0.6%) and three related to the absence of spleen (1.9%) with septic shock as manifestation. CONCLUSIONS: The complications related to splenectomy in the Sugiura-Futagawa procedure are low, as well as the overwhelming post-splenectomy infection rate. Splenectomy in the Sugiura-Futagawa operation has no important role in the post-operative morbidity. PMID- 7481448 TI - [Dynamic high-contrast computed tomography in the diagnosis of pancreatic necrosis]. AB - This work intended to find out whether dynamic high-contrast computed axial tomography (CAT scan) in effective and more useful than conventional computed tomography for the diagnosis of pancreatic necrosis in patients with severe acute pancreatitis. BACKGROUND: Although many methods have been used to predict the severity and extent of pancreatic necrosis, few studies have assessed computed tomography. METHODS: Longitudinal, prospective and comparative study was performed on consecutive patients with severe acute pancreatitis in which dynamic high-contrast computed tomography and conventional computed tomography were carried out. RESULTS: In pancreatic necrosis, pancreatic densities were lower. Necrosis by itself was also associated with higher severity scores, but not with a higher mortality rate. However, mortality rates were influenced by the extent of pancreatic necrosis. Sensitivity of 100%, specificity of 68%, positive predictive and negative predictive values of 100 and 42% were achieved. STATISTICS: Densities of TCNL and TCAC were analyzed using student's t test of independent samples. The correlation of the aorta/pancreas index and the presence of necrosis was analyzed using a linear correlation test. CONCLUSIONS: The authors conclude that dynamic high-contrast computed tomography is more effective than conventional computed tomography in diagnosing pancreatic necrosis. Moreover, the diagnosis of pancreatic necrosis and its extent makes it possible to formulate a prognosis as to severity of the disease. PMID- 7481449 TI - [Duodenal cancer]. AB - To analyze the general characteristics and outcome of a cohort of 6 patients with malignant duodenal tumors who underwent surgical radical resection. DESIGN: Longitudinal retrolective study. SETTING: A third level medical center. RESULTS: Mean age was 48 years. M/F ratio was 2:1. The most frequent symptom was duodenal obstruction. Mean time between the beginning of the symptoms and the correct diagnosis was 15 months. Five of six patients demonstrated abnormalities on gastrointestinal X rays. All patients underwent Whipple resection, this was curative in 2, palliative in 2 more and not evaluated in two cases due to operative mortality. Five tumors were adenocarcinomas and one a carcinoid tumor. The long-term mortality occurred in 2 patients. CONCLUSIONS: The diagnosis of primary duodenal carcinoma needs a deep clinical suspicion, some patients when treated by radical resection may experience long-term survival. PMID- 7481450 TI - [Cyto-reduction of rectal cancer after preoperative chemoradiotherapy]. PMID- 7481451 TI - [Pathogenesis of cirrhotic hepatic encephalopathy. Treatment implications]. AB - BACKGROUND: Hepatic encephalopathy (HE) is a neuropsychologic syndrome associated with multifactorial metabolic disruptions. Several physiopathogenic theories have been proposed. OBJECTIVES: To review the most important theories (ammonia, mercaptans, short chain fatty acids, false neurotransmitters, gamma-aminobutyric acid, endogenous benzodiazepines) and to analyze the different therapeutic modalities derived from these theories. METHODS: An intensive bibliographic review was carried out using Med-Line and Current Contents data base. Randomized therapeutic trials published in the last ten years were particularly analyzed. RESULTS: From all theories proposed, the revised ammonia theory better explains pathogenesis of HE based on the information about efficacy of anti-ammonia agents. Authors propose a theory in order to explain the effect of ammonia and serotonin in neurotransmission, particularly excitatory glutamatergic neurotransmission. CONCLUSIONS: It seems that ammonia has a predominant role in pathogenesis of HE. Some other agents may be involved but its nature is not actually defined. Glutamatergic neurotransmission un HE has been recently studied. PMID- 7481453 TI - [Epidemiology of digestive system cancer in the State of Sonora]. PMID- 7481454 TI - [Internal biliary fistula: study of 13 cases and review of the literature]. AB - Between 1986 and 1994 13 patients with spontaneous internal biliary fistula were operated at the Hospital das Clinicas of the University of Sao Paulo. Ten were patients women and three were men. Their mean age was 55.2 years (range 30 to 87 years). The etiology was cholelithiasis in all cases and the most frequent type of fistula was cholecystoduodenal. There was no mortality. A review of literature of this condition and a discussion about the diagnosis, treatment and etiopathogenesis are presented. PMID- 7481455 TI - [Hepatic venous system and its application in the so-called split-liver transplantation technic]. AB - Sixty fresh adult livers were obtained from cadavers together with celiac trunk, head of the pancreas and superior mesenteric artery. The right portal vein, left portal vein and their respective branches were dissected as well as the hepatic veins. There was only one right hepatic vein in 59 cases. The median hepatic vein was present in 53 (88.3%) cases and the left hepatic vein only in 46(76.3%). In 59(98.3%) cases, there were right and left portal vein but in one (1.6%) case no portal bifurcation has been found. The median portal vein has been found only in 9(15.2%) cases. PMID- 7481452 TI - [Hereditary nonpolyposis colorectal cancer. Lynch syndrome]. AB - BACKGROUND: Hereditary non-polyposis colorectal cancer represent 5% of the colorectal cancer population. It's characterized by an autosomal dominant inheritance; there are two variants: Lynch syndrome I is characterized by predisposition to early onset colorectal cancer, with predominance for proximal location to the splenic flexure and synchronous and/or metachronous colorectal cancers. Lynch syndrome II shares the same features, but shows association with extracolonic cancers, particularly carcinomas of endometrium, ovary and stomach. However, this syndrome is not easily recognized, due to the lack of phenotypical expression as familiar adenomatous polyposis. OBJECTIVE: The purpose of this report is to review the natural history, diagnosis, treatment, surveillance of the probands and their families, and to update recent research on the genotypic risk biomarkers located in chromosomes 2, 3 and 7. METHODS: We analyzed the available literature to discuss the clinical characteristics, pathology, genetics, management, and surveillance of hereditary non-polyposis colorectal cancer. CONCLUSIONS: The diagnosis of hereditary non-polyposis colorectal carcinoma is not a rare event. It requires the demonstration of vertical transmission, therefore, the past history of colorectal cancer in the proband and his family should be carefully taken; including the anatomic site, histology and age at diagnosis. PMID- 7481456 TI - [Infections of central venous catheters]. AB - The central venous catheter is considered the lifeline of patients requiring long term parenteral nutrition, and its infectious complications accordingly represent a frequent cause for hospitalization and morbidity. Modern catheters made of Silastic increase the duration and efficiency of venous access, but their substitution is also more complex and expensive. In a small series of patients undergoing home parenteral nutrition due to short bowel syndrome, and carrying. Silastic catheters, infection of the cannula was treated by administration of systemic as well as local antimicrobial agents. The septic process was resolved without the need of removing the catheters, and with excellent tolerance to the medications. It is concluded that in-situ sterilization of long-term catheters should be attempted in selected cases. PMID- 7481457 TI - [Experimental evaluation in mice of vitamin B complex repellent activity against Culex quinquefasciatus]. AB - One untested but widely held opinion in Brazil is that oral use of vitamin B complex is useful as a mosquito repellent. We exposed Balb-C mice to a large number of Culex quinquefasciatus females, after giving vitamin B complex to mice (drops by mouth). There was no difference between mice attractiveness to mosquitoes in the vitamin B group compared to normal, but very few of the females bite mice in both groups, so the experiment was inconclusive. We suggest further experiments in this subject, using other models were the animals are more attractive to mosquitoes. PMID- 7481459 TI - Intestinal transit and opaque enema study in chagasic mice. AB - The report concerns as experimental study utilizing thirty-three mice inoculated subcutaneously with 1000 blood forms of Trypanosoma cruzi. Intestinal transit and morphology of colon were studies by means of X-rays. In subacute stage intestinal transit was normal. In chronic stages the mice displayed a delay in evacuation time. In one out of 12 animals the opaque enema documented the existence of megacolon. PMID- 7481458 TI - [Risk factors for surgical wound infection development in head and neck cancer surgery]. AB - Over a period of 24 months, patients undergoing surgical procedures for head and neck cancer at the Cancer Hospital in Rio de Janeiro, Brazil were followed-up prospectively for identification of surgical wound risk factors. A total of 273 patients fulfilling the National Nosocomial Infection Surveillance (NNIS) surgical criteria completed the analysis. The overall wound infection rate was 25.3%. Of the 11 potential risk factors for postoperative wound infections analysed by stepwise multiple logistic regression, the five variables that were independent of each other and highly predictive were patient undergoing an operation classified as either contaminated or infected, a surgery lasting longer than 5 hours, an American Society of Anesthesiologist (ASA) preoperative assessment score of 4 and 5, age over 50 years and prior radiotherapy. The preoperative stay, antibiotic prophylaxis, surgical drains, preoperative nutritional status, sex and the presence of prior infection did not have an independent significant contribution to the risk of infection. This study reports specific risk factors for head and neck cancer surgeries and contributes substantially to the effectiveness of infection control strategies to prevent their occurrence in this high-risk group of patients. PMID- 7481462 TI - [Participation of the adhesion molecules in the development of inflammatory response]. AB - The authors present a review of adhesion molecules involved in inflammatory response. The recent description of adhesion molecules expressed on circulating leukocytes and endothelial cells, have been elucidated the leukocyte-endothelial interactions, essential for transmigration of leukocytes into tissues. It also indicates how anti-inflammatory drugs affect adhesion molecules. Finally, anti adhesion therapies are discussed as new strategies for the treatment of inflammatory diseases. PMID- 7481461 TI - [Cardiovascular impact of alcoholism]. AB - A history of alcohol abuse associated with hypertension and chronic alcohol consumption is the leading cause of secondary cardiomyopathy. Both acute and chronic alcohol consumption precipitate arrhythmias. However moderate alcohol intake exerts a protective effect against coronary heart disease and stroke. Morbidity is lowest among light drinkers, slightly higher among lifelong abstainers and much higher among heavy drinkers. Magnesium deficiency is common among chronic alcoholics and frequent in Brazil. Magnesium therapy may have beneficial effects when there is excessive ethanol consumption. PMID- 7481460 TI - [Nutritional and energy metabolism changes in abdominal irradiation--experimental study]. AB - In this study the effects on nutritional status and energetic metabolism due to abdominal irradiation were analysed. Adult male Wistar rats (48), were divided in two groups Control (C) and Radiated (R). The rats were maintained all time in metabolic cages. The study was done in two periods: Period 1 begun at 0 day, where rats adapted to cages and oral diet, had food and water "ad libitum". At the day 4 indirect calorimetric measurements were performed (calorimetry I). At Period 2, group R rats abdominal radiation at a 300cGy/day rate, for 5 consecutive days, and group C started a pair feeding process linked individually to R rats and suffered application of simulated-radiation. Two other calorimetric measurements (II,III) were performed during Period 2. After radiation the last calorimetry was performed (IV). At sacrifice (day 14) blood was collected for determination of hemoglobin, haematocrit, albumin and transferrin. There were no statistical differences among groups C and R during Period 1 (p < 0.05). Great reduction in food intake and weight variation were found in Period 2, but weight loss was significantly higher in R rats. Nitrogen balance decreased in Period 2, but without difference among the groups (p < 0.05). Serum albumin was significantly lower in R rats. Respiratory quotient decreased in both groups during Period 2, but R rats kept it lower (p < 0.05). The energy expenditure level decreased after radiation in Group R. During Period 2 total substrate oxidation decreased in R rats. Radiation decreased glucose and protein oxidation.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7481463 TI - [Medical staff of teaching hospitals and the medical education process]. AB - At the in S. Paulo University Medical School (FMUSP), 39% of physicians that work there take some part in teaching work ten percent of these professionals had completed the mastership, 25% concluded the doctorship and 5% are free professors. PMID- 7481464 TI - Chagas' disease in the Brazilian Amazon. II. A serological survey. AB - A serological survey, involving indirect immunofluorescence testing of blood sera samples, was carried out on the residents of one in every five dwellings in the town of Barcelos (in the northern part of the State of Amazonas, on the right bank of the Rio Negro, 490 Km from Manaus by river) and on the rural populations of the villages of Piloto and Marara (also on the right bank of the Rio Negro, 30 minutes by boat from Barcelos). A total of 710 sera samples were tested, 628 from the resident population in the town of Barcelos, 35 from Piloto and 47 from Marara. The tests were carried out using human anti-gammaglobulin type IgG (Biolab) and antigen from formolized culture of T. cruzi Y strain. The sera were serially diluted from 1:40 to 1:320 in PBS 7.2. Of the 710 samples examined 89 (12.5%) were positive for anti-T.cruzi antibodies: 2 of these (2.2%) at a dilution of 1:320; 12 (13.4%) at 1:160; 38 (42.6%) at 1:80; and the remainder at 1:40, giving a median serological dilution of 1:80. The following questions are discussed: the high serological prevalence for Chagas' infection found in our survey; the possibility of serological cross-reactions; the need for confirmatory tests for the positives reactions; and the strong correlation between our results and preliminary epidemiological data (such as the level of human contact with wild triatominae, know locally as "Piacava's lice". We draw attention to the isolation by xenodiagnosis of one strain of T.cruzi from a patient with positive serology for Chagas' infection. PMID- 7481465 TI - Dot-ELISA for the detection of IgM and IgG antibodies to Schistosoma mansoni worm and egg antigens, associated with egg excretion by patients. AB - Human schistosomiasis, caused by Schistosoma mansoni, is highly prevalent in Brazil and usually diagnosed by time consuming stool analysis. Serological tests are of limited use in this disease, mainly for epidemiological studies, showing no discrimination between previous contact with the parasite and active infections. In the present study, we standardized and compared a Dot-ELISA for IgM and IgG antibodies against S. mansoni antigens from eggs and worms with a routine IgG and IgM immunofluorescence assay using similar antigens, in the study of sera from 27 patients who had quantified egg stool excretion. The positivity obtained for IgG Dot-ELISA was 96.3% and 88.9% for IgM Dot-ELISA with worm antigen and 92.6% and 90.9% with egg antigen. The IFI presented similar positivities using worm antigen, 92.6% (IgG) and 96.3% (IgM), and lower results with egg antigen, 77.8%(IgG and IgM). The patients studied were divided into two groups according to their egg excretion, with greater positivity of serological tests in higher egg excreters. When comparing the quantitative egg excretion and the serological titers of the patients, we detected a correlation only with IgM Dot-ELISA, with r = 0.552 (p = 0.0127). These data show that Dot-ELISA can be used for the detection of specific antibodies against S. mansoni in sera from suspected patients or in epidemiological studies and, with further purification of egg antigen and larger samples, IgM Dot-ELISA could be a possible tool for rough estimates of parasite burden in epidemiological studies. PMID- 7481466 TI - Diagnosis of human toxoplasmosis by a Dot enzyme-linked immunosorbent assay. AB - A Dot enzyme-linked immunosorbent assay (Dot-ELISA) was standardized and evaluated for the serodiagnosis of human toxoplasmosis. Out of 538 serum samples tested by the immunofluorescence test for toxoplasmosis (IFAT-IgG) as reference test, 183 (34%) were positive at cut off 1:16 and 192 (36%) were positive for Dot ELISA-IgG at cut-off 1:256. For Dot-ELISA, co-positivity was 0.94, co-negativity 0.94 and concordance 0.88 in relation to IFAT-IgG. These results suggest the usefulness of Dot-ELISA (cut-off titer of 1:256) for the serodiagnosis of human toxoplasmosis. The main advantage of this technique is simplicity, positive test can be visually identified (colored precipitate). It does not require a special equipment and it can be used as a qualitative test to screen large numbers of samples or as a quantitative assay to determine end-point titration of individual sera. PMID- 7481467 TI - Evaluation of the enzyme-linked-immuno-electro-diffusion-assay (ELIEDA) for the diagnosis of Schistosoma mansoni infection with low worm burden. AB - An immunoprecipitation technique, ELIEDA (enzyme-linked-immuno-electro-diffusion assay), was evaluated for the diagnosis of Schistosoma mansoni infection with low worm burden. One hundred of serum samples from patients excreting less than 600 eggs per gram of feces (epg), with unrelated diseases and clinically healthy subjects were studied. In patients with egg counts higher than 200 epg, the sensitivities of IgM and IgG ELIEDA were 1,000 and 0.923, respectively, not differing from other serologic techniques, such as indirect hemaglutination (IHAT), immunofluorescence (IFT) tests and immuno-electrodiffusion assay (IEDA). However in patients with low egg counts (< 100 epg), the IgG ELIEDA provided better results (0.821) than IgM ELIEDA (0.679), showing sensitivity that did not differ from that of IgG IFT (0.929), but lower than that of IgM IFT (0.964). However, its sensitivity was higher than that found with IHAT (0.607) and IEDA (0.536). The specificity of IgG ELIEDA was comparable to that of other techniques. The data indicate that IgG ELIEDA might be useful for the diagnosis of slight S. mansoni infections, and the cellulose acetate membrane strips can be stored for further retrospective studies. PMID- 7481468 TI - Natural killer cell activity in experimental paracoccidioidomycosis of the Syrian hamster. AB - The study evaluated the activity of NK cells during the course of experimental infection of hamsters with Paracoccidioides brasiliensis. Eighty hamsters were infected with P. brasiliensis by intratesticular route and sacrificed at 24h, 48h, 96h, 1, 2, 4, 8 and 11 weeks of infection and compared to 40 noninfected hamsters employed as controls. These animals were submitted to the study of NK cytotoxic activity by a single-cell assay and humoral immune response by immunodiffusion and ELISA tests. The production of macrophage migration inhibitory factor in the presence of Phytohemagglutinin and P. brasiliensis antigen and histopathology of the lesions were evaluated at 1, 4, 8 and 11 weeks of infection. The infected animals displayed significantly high levels of NK activity during the four weeks of infection that decreased from the 8th week on when compared to controls. This impairment of NK activity was associated with depression of cell-mediated immune response and with increase in the extension of the histopathologic lesions. There was an inverse correlation between NK cell activity and specific antibody levels. The results suggest that after initial activation, NK cells were unable to control the fungus dissemination. The impairment of NK activity in the late stages of the infection might be related to immunoregulatory disturbances associated with paracoccidioidomycosis. PMID- 7481469 TI - Central nervous system virion detection in acute measles: histopathological, ultrastructural and pathogenetic aspects. AB - Histopathological and ultrastructural studies of 23 patients who died with clinical diagnosis of measles were carried out. In 12 cases viral nucleocapsids were searched by electron microscopy and detected in 100% of the cases in the lungs and in 50% of the cases in the central nervous system. They were mostly intranuclear. Histopathological changes associated to neurological alterations and the detection of virion are discussed in relation to acute and delayed clinical manifestations. PMID- 7481471 TI - Changes in pentobarbital induced sleeping-time in mice infected with Schistosoma mansoni and immunosuppressed. AB - To evaluate whether the intensity of the hepatic granulomatous response induced by S. mansoni eggs plays a role in drug metabolism, mice were infected with 40 cercariae and tested to assess the sodic pentobarbital induced sleeping-time. To decrease the inflammatory reaction the animals were irradiated with 400 Rad or received azathioprine, 20 mg/kg, 3 times a week, for 4 weeks, respectively in or beginning in the 33th post-infection day. In infected animals receiving azathioprine the area of the hepatic granulomas was smaller and the sleeping-time was similar to that of non-infected ones (controls). In mice infected and irradiated the granuloma dimensions were similar to those of animals only infected, in these two latter groups of animals, the sleeping-time was more prolonged than that of the control animals. These results show that: 1) mice with unaltered hepatic granulomatous reaction show reduction in metabolism of sodic pentobarbital; 2) granulomatous response diminished by azathioprine does not interfere with the capacity of metabolism of the anesthetic drug. PMID- 7481470 TI - Intestinal parasitism in the Xavante Indians, central Brazil. AB - This paper reports the findings of a survey for intestinal parasites among the Xavante Indians from Central Brazil. A. lumbricoides (25.0%) and hookworms (33.6%) were the two most common helminths; E. histolytica complex (7.8%) and G. lamblia (8.6%) the most common protozoans. The majority (58.5%) of positive individuals hosted only one species of helminth. Egg counts for helminths, and for A. lumbricoides in particular, were found to be not dispersed at random, with a few individuals, all of whom young children, showing very high counts. The prevalence rates of intestinal parasites for the Xavante are below those reported for other Amerindian populations from Brazil. PMID- 7481472 TI - Cardiac sympathetic-parasympathetic balance in rats with experimentally-induced acute chagasic myocarditis. AB - To clarify the mechanism responsible for the transient sinus tachycardia in rats with acute chagasic myocarditis, we have examined the cardiac sympathetic parasympathetic balance of 29 rats inoculated with 200,000 parasites (Trypanosoma cruzi). Sixteen infected animals and 8 controls were studied between days 18 and 21 after inoculation (acute stage). The remaining 13 infected animals and 9 controls were studied between days 60 and 70 after inoculation (sub-acute stage). Under anesthesia (urethane 1.25 g/kg), all animals received intravenous atenolol (5 mg/kg) and atropine (10 mg/kg). Acute stage: The baseline heart rate of the infected animals was significantly higher than that of the controls (P < 0.0001). The magnitude of the negative chronotropic response to atenolol was 4 times that of the controls (P < 0.00001). This response correlated with the baseline heart rate (r = -0.72, P < 0.001). The heart rate responses to the beta-blocker and to atropine, of the infected animals studied during the sub-acute stage, were not different from controls. These findings suggest that cardiac sympathetic activity is transiently enhanced and cardiac parasympathetic activity is not impaired, in rats with acute chagasic myocarditis. The transient predominance of cardiac sympathetic activity could explain, in part, the sinus tachycardia observed in the acute stage of experimentally-induced chagasic myocarditis. PMID- 7481474 TI - Mammomanogamus (Syngamus) laryngeus infection: a new Brazilian human case. PMID- 7481473 TI - The experimental granuloma. A hypothesis to explain the persistence of the lesion. AB - Granulomatous inflammation is the morphological substrate of a variety of important infectious diseases such as tuberculosis, leprosy, schistosomiasis and others. Nevertheless, although many aspects of this special type of inflammation are known, fundamental questions concerning granuloma formation, persistence, fate and significance for host-parasite relationships still remain to be elucidated. In this brief review, the basic and more relevant literature related to experimental investigations on granuloma physiopathology is presented. Based on recent investigations performed in our laboratory showing that MDF (Macrophage Deactivating Factor) secreted by epithelioid cells and characterized as the calcium-binding protein protein MRP-14 deactivates activated macrophages, a hypothesis to explain the persistence of granulomatous inflammation is put forward. PMID- 7481476 TI - Geographical considerations on HAM/TSP in Japan. PMID- 7481475 TI - A case of mycotic keratitis caused by Fusarium solani. AB - A 36-year-old black man, without history of systemic disease or ocular trauma developed a corneal infection in his left eye. He was treated with antibacterial antibiotic and corticosteroids for one month prior to diagnosis. Fungal hyphae and chlamydospores were found in a KOH preparation of the corneal scrapings, and positive cultures for Fusarium solani were obtained in Sabouraud dextrose agar. It is emphasized the cautious use of antibiotics and steroids in corneal diseases, and the need of considering the involvement of opportunistic fungi in the etiology of these infections. PMID- 7481477 TI - Mycobacterium avium complex (MAC): an unusual potential pathogen in cerebrospinal fluid of AIDS patients. AB - Mycobacterium avium complex (MAC) is frequently isolated from patients with late complications of Acquired Immunodeficiency Syndrome (AIDS), especially in North America and Europe. However, its isolation from the central nervous system (CNS) has been seldom reported in these countries. MAC infections in AIDS patients in African and Latin American countries are believed to be uncommon. We report the isolation of MAC from cerebrospinal fluid (CSF) of 11 AIDS patients out of 1723 (0.63%) seen at "Centro de Referencia e Treinamento-AIDS", Sao Paulo and discuss the significance of its isolation. PMID- 7481478 TI - Blood culture as a parameter of treatment effectiveness in experimental histoplasmosis of the hamster. AB - The aim of this study was to determine the value of blood culture as a parameter of treatment effectiveness in experimental histoplasmosis. A total of thirty five hamsters, weighing approximately 120g, were inoculated intracardially with 0.1 ml of a suspension containing 4 x 10(7) cells/ml of the yeast phase of H. capsulatum. Treatments were started one week after the infection and lasted for 3 weeks. The azoles, (itraconazole, saperconazole and fluconazole) were administered once a day by gavage, at a dose of 8 mg/kg; Amphotericin B was given intraperitoneally every other day at a dose of 6mg/kg. Blood samples (1 ml) were obtained by heart punction from the 4th day after infection and were seeded in Sabouraud honey-agar and BHI-agar. The hamsters that survived were killed one week after treatment completion and the following criteria were considered for treatment evaluation: 1) rate of spontaneous death, at the end of the experience; 2) microscopic examination of Giemsa smears from liver and spleen and 3) determination of CFU in spleen cultures. Amphotericin B was the most effective drug, with negative blood cultures at day 20, negative spleen cultures in all cases and all the animals survived until the end of the study. Fluconazole was the less effective drug, blood cultures were positive during the whole experience, spleen cultures showed a similar average of CFU when compared with the control animals and 42.8% of these animals died. Saperconazole and itraconazole showed a similar activity, with survival of all hamsters and negative blood cultures at 23 and 26 days respectively. Blood culture seems to be valuable parameter for treatments' evaluation in experimental histoplasmosis of the hamster. PMID- 7481479 TI - Biochemical analysis of ferritin subunits in sera from adult Still's disease patients. AB - To determine the origin of the increased serum ferritin that occurs in adult Still's disease (ASD), we analyzed subunits of the serum ferritin as follows. Gel filtration with Sepharose CL-6B demonstrated that the molecular weight of serum ferritin was about 490 kDa. Western blot analysis revealed only L-subunits (molecular weight 19 kDa) in patients with serum ferritin levels higher than 1,000 ng/ml. Patients with serum ferritin levels higher than 1,000 ng/ml, however, showed G-subunits (molecular weight 23 kDa) in addition to the L subunits. When concanavalin A (Con-A) Sepharose 4B was used in an absorption test, the percentage absorption was extremely low in the patients with serum ferritin levels higher than 1,000 ng/ml. Isoferritin patterns of the patients determined by chromatofocusing revealed traces of acidic ferritin. The findings suggested that glycosylated ferritin does not account for the major portion of the increased serum ferritin. PMID- 7481480 TI - An immunohistochemical study of immunological phenomena in minor salivary glands in patients with Sjogren's syndrome. AB - Using different monoclonal antibodies, we performed an immunofluorescent technique on labial salivary glands in order to investigate the immunological phenomena involved in Sjogren's syndrome (SS). An aberrant expression of HLA-DR molecules was detected on cytoplasm of epithelial labial salivary cells in 9 out of 19 (47%) patients, with SS. No such expression was found in 8 patients without SS or in 3 normal controls. HLA-DQ molecules were demonstrated also in two out of ten SS patients without HLA-DR. A lymphocytic infiltration was not correlated with the expression of class II molecules. T cells bearing gamma delta receptors were not detected. The intracellular adhesion molecules (ICAM-1) and lymphocyte function associated antigen-1 (LFA-1) were not found on epithelial glandular salivary cells of patients and controls. In conclusion, these data suggested that the absence of ICAM-1 and LFA-1 in salivary cells and the absence of infiltrating T cells bearing gamma delta receptors exclude their immunopathogenetic role in SS; moreover, these data demonstrated that the aberrant expression of HLA class II molecules on epithelial salivary cells of patients with SS is not a phenomenon correlated with the lymphocytic infiltration. PMID- 7481482 TI - Spa therapy for gonarthrosis: a prospective study. AB - The objective of this study was to evaluate the effect of spa therapy on clinical parameters of patients with gonarthrosis. Patients with gonarthrosis (n = 33) underwent a 2-week spa therapy using three treatment regimes and a 20-week follow up as follows: group I (n = 11) had mineral water baths and hot native mineral mud packs, group II (n = 12) had mineral water baths and rinsed mineral-free mud packs and group III (n = 10) had tap water baths and mineral-free mud packs. The patients and the assessing rheumatologist were blinded to the difference in the treatment protocols. A significant improvement in the index of severity of the knee (ISK), as well as night pain scores, was achieved in group I. Improvement in physical findings and a reduction in pain ratings on a visual analogue scale (VAS) did not reach statistical significance. Analgesic consumption was significantly decreased in both groups I and III for up to 12 weeks. Global improvement assessed by patients and physician was observed in all three groups up to 16 weeks but persisted to the end of the follow-up period in group I only. Patients with gonarthrosis seemed to benefit from spa therapy under all three regimes. However, for two parameters (night pain and ISK) the combination of mineral water baths and mud packs (group I) appeared to be superior. PMID- 7481481 TI - Longitudinal study on the production of and cellular response to interleukin-2 in patients with systemic lupus erythematosus. AB - Interleukin-2 (IL-2) has been proven to be a defective element in immune regulation in systemic lupus erythematosus (SLE). However, its course in time is unknown. We studied its production and cellular response in the peripheral blood cells of 30 SLE patients and 12 healthy subjects. In addition, we studied the spontaneous and lipopolysaccharide (LPS) induced production of IL-1, which have been found to be, respectively, increased and lowered in untreated SLE patients. Patients were studied at the outset, when still untreated, and at 1, 2, 6, 12, 18, and 24 months. At the outset, 18 had active disease and 12 were in remission. The decreased proliferative response of T cells to IL-2 and the deficient production of IL-1 upon LPS induction became normal after 6 months treatment, whereas the expression of high affinity IL-2 receptors took 18 months to become normal and the deficient production of IL-2 took 2 years. Despite clinical remission, the decreased capacity of T cells to absorb IL-2 persisted for 2 years. The effect of various prednisone dosages on the measured variables was evaluated. With intermediate doses of prednisone (20-45 mg), we observed the largest improvement in IL-2 production and in IL-1 production upon LPS stimulation. Higher doses of prednisone reduced also the spontaneous production of IL-1 and resulted in an increase in the expression of CD25+ cells. The addition of low doses of cytotoxic drugs (oral cyclophosphamide or azathioprine) resulted in an improvement in the capacity to absorb IL-2 and a reduction in spontaneous IL-1 production.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7481483 TI - In vivo and in vitro expression of adhesion molecules by peripheral blood lymphocytes from patients with primary Sjogren's syndrome: culture-associated enhancement of LECAM-1 and CD44. AB - The interaction of adhesion receptors on lymphocytes with their ligands over endothelial cells provides the mechanism by which lymphocytes infiltrate target tissues in autoimmune diseases. Primary Sjogren's syndrome (SS) is associated with lymphocytic infiltration in exocrine glands. The aim of this study was to examine levels of expression of adhesion molecules by peripheral blood lymphocytes from patients with SS (before and after stimulation). Peripheral blood lymphocytes from 16 patients with primary SS and from 15 controls were stained directly or cultured for 72 h with and without phytohaemagglutinin (PHA). Indirect immunofluorescence with monoclonal antibodies and flow cytometry were used. The following molecules were detected in patients before culture: CD18 (mean percentage 94%), CD11a (94%), CD11b (39%), CD54 (23%), CD58 (62%), CD44 (Hermes-1; 82%), CD49-d (VLA-4; 80%), CD25 (11%) and LECAM-1 (62%). After stimulation with PHA, there was an increase in the levels of CD18 (2.5-fold), CD11a (2.3-fold), CD54 (10.2-fold), CD58 (2.5-fold), CD44 (2.4-fold), CD49d (3.4 fold) and CD25 (62-fold) on lymphocytes from both patients and controls. The number of positive cells and level of expression did not differ from the controls, except in the case of unstimulated, cultured lymphocytes in which the levels of CD44 and LECAM-1 were increased more in patients than in normal controls. The increase in the level of in vitro expression of CD44 (P < 0.05) and LECAM-1 (P < 0.002) on lymphocytes from patients with primary SS reached statistical significance when compared to similarly cultured lymphocytes from controls.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7481484 TI - Elevated levels of soluble intercellular adhesion molecule-1 correlate with disease activity in Behcet's disease. AB - The objective of this study was to measure soluble intercellular adhesion molecule-1 (sICAM-1) in patients with Behcet's disease (BD) and to analyse the relationship of sICAM-1 levels with clinical and some laboratory measures of disease activity. Forty patients with BD fulfilling the International Study Group Criteria for the diagnosis of BD and 20 healthy controls were studied. Twenty patients had active, and 20 patients had inactive disease. Serum sICAM-1 was determined by a sandwich ELISA. The mean (+/- SD) sICAM-1 level was significantly higher in the whole BD group (297.3 +/- 86.6 ng/ml) than in the healthy controls (213 +/- 83.5 ng/ml; P < 0.05). The mean sICAM-1 levels in active and inactive BD patients were 315.7 +/- 76.3 ng/ml and 258.3 +/- 73.3 ng/ml, respectively. The mean sICAM-1 level in active patients was significantly higher than in inactive patients and healthy controls (P < 0.02 and P < 0.001, respectively). No statistically significant difference in mean sICAM-1 levels was found between inactive BD patients and healthy controls (P > 0.05). There was no statistically significant difference between the mean sICAM-1 levels of active patients with (351.3 +/- 77.2 ng/ml) or without vascular lesions (292 +/- 68.8; P > 0.05). In spite of a positive correlation between disease activity and both erythrocyte sedimentation rate and C-reactive protein (CRP; P < 0.01), we found no correlation between sICAM-1 and either of them (P > 0.05). The elevated levels of sICAM-1 may be due to endothelial cell activation and/or damage or may be the result of inflammation.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7481485 TI - The specificity of the anti-Proteus antibody response in tissue-typed rheumatoid arthritis (RA) patients from Brest. AB - Anti-Proteus mirabilis antibody titres were found to be elevated in 50 active tissue-typed French rheumatoid arthritis (RA) patients from Brest when compared to 49 healthy French controls using enzyme-linked immunosorbent assay (ELISA; P < 0.001) and indirect immunofluorescence assay (IIFA; P < 0.001). However, there was no significant elevation in antibody titres against Escherichia coli or Salmonella typhimurium in the RA patients compared to the controls when measured by ELISA. Serum levels of C-reactive protein (CRP) were also found to be significantly higher in RA patients when compared to healthy control subjects (P < 0.001). These results suggest that P. mirabilis may play an important and specific role in the triggering and persistence of RA. PMID- 7481487 TI - Collaborating without caving in. PMID- 7481486 TI - Expression of a multidrug resistance gene in human rheumatoid synovium. AB - The objective of this study was to assess the expression of a multidrug resistance (MDR) phenotype, implicated in the cellular resistance of tumor to chemotherapy, in rheumatoid synovial membrane. Synovial membrane from 16 rheumatoid (RA) patients was studied. Six patients with osteoarthritis constituted the control group. The cell membrane expression of the glycoprotein Pgp 170, encoded by the MDR 1 gene, was determined by an immunoperoxidase technique using two different monoclonal antibodies (JSB 1, C 219). The polymerase chain reaction (PCR) methods were used in parallel to detect the presence of the MDR 1 gene mRNA in the synovial cells. Pgp 170 was expressed on the cell membrane of five RA patients and MDR 1 cellular transcription was detected in one other RA patient. We did not observe any association between synovial glycoprotein expression and age, disease activity, and a specific treatment with a long-acting drug. However, MDR protein expression was associated with the successive treatment with more than three disease-modifying antirheumatic drugs (DMARDs). We concluded that the synovial membrane expresses a glycoprotein recognized by the antibodies JSB 1 and C 219. The absence of concomitant MDR 1 transcription suggests the expression of an atypical MDR phenotype in the synovial membrane, distinct from the Pgp 170 encoded by the MDR 1 gene. The implications of the MDR phenotype and the resistance of RA to DMARDs is further discussed. PMID- 7481488 TI - When bone disappears. PMID- 7481489 TI - Who says there's nothing we can do? PMID- 7481490 TI - Wide-complex tachycardia. PMID- 7481491 TI - Diabetes 2000. Complications during pregnancy. PMID- 7481492 TI - 1995 earnings survey: the news is mixed. PMID- 7481493 TI - From hospital to hospice: bridging the gap. PMID- 7481494 TI - Once the patient's home. PMID- 7481495 TI - Serving as an expert witness. PMID- 7481496 TI - BSN again? PMID- 7481498 TI - Help for spouses of chronically ill patients. PMID- 7481497 TI - The risk of cross-sensitive drug reactions. PMID- 7481499 TI - [Cadmium content in daily food rations of rural school-age children]. AB - Cadmium was determined in 230 daily food rations of children aged 8-9 years and 12-13 years. The study was carried out in spring and in autumn during three successive days. Daily meals were collected from the households selected in the area of five villages. Cadmium was determined by the extraction ASA method, after dry mineralization at about 450 degrees C. The level of cadmium in the examined food rations varied from 6 to 569 micrograms/person/day, with the mean value of 43.5 +/- 52.6 micrograms/person/day, and the median of 31.3 micrograms. In 93% of food rations the amount of cadmium was below the upper limit--the dose permitted by the WHO for adults i.e. 57-71 micrograms/day. Considering the children's smaller body weight the daily cadmium intake in food rations was converted into the daily cadmium intake per 1 kilogram body weight. In the group of children aged 8-9 years cadmium intake was 0.3-15 micrograms, with the mean value of 1.7 micrograms, and in the group of 12-13-years-old children it was 0.1-13.9 micrograms, with the mean of 1.3 micrograms. Only 36% of food rations in the group of younger children and 58% of food rations in the older group contained the amount of cadmium below the tolerable value of 1 microgram/kg body weight/day. The weekly cadmium intake was calculated for each child based on its amount in daily food rations during the successive three days. The mean value was 12.3 micrograms/kg body weight, with the median of 8.1 micrograms, within the range of values 2.1-58.8 micrograms.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7481503 TI - [HACCP--system of analysis to determine critical control points to assure food safety]. AB - The philosophy and application of HACCP system in food processing, catering and food inspection to ensure food safety has been presented. The HACCP system stages were pointed out and explained. Also the advantageous and difficulties in implementation of HACCP system were presented. PMID- 7481502 TI - [Determination of lead (II) in food using the ICP-AES method after condensation on the lanthanum carrier]. AB - The purpose of the study was utilization of emission atomic absorption spectrometry with excitation in induction-coupled plasma, and molecular spectrometry for determination of lead in milk and dairy products after condensation in ammonia atmosphere. Two methods were proposed for lead condensation on lanthanum carrier--in form of hydroxide or sulphide. By coprecipitation of lead on lanthanum in ammonia atmosphere lead separated and selectively condensed. Other metals (Zn, Cd, Cu) coexistent in food products remained as amino complexes in the solution. The sediment remaining after lead condensation was dissolved in 2 mol/dm3 hydrochloric acid solution. The selected conditions of lead separation and condensation make possible determination of the metal in the concentrations from 0.01 mcg/ml to 0.2 mcg/ml by the ICP-ARS and dithizone methods. The method of condensation and determination was used for lead determination in milk and dairy products in concentrations below 1.5 x 10(-5)%. The studied food products were mineralized with concentrated HNO3, H2SO4 and perhydrol. This mineralization method makes it possible to determine in one mineralisate directly copper and zinc by ICP-ARS and dithizone methods, and lead after condensation. The obtained results were precise and reproducible. PMID- 7481500 TI - [A study of the content of synthetic food dyes in gelatin jellies]. AB - Methods of identification and quantitative determination of synthetic dyes in gelatine jellies have been described. Extraction and isolation procedures of synthetic food dyes have been developed specifically for gelatine products. The various techniques have been used as follows: paper chromatography, absorption spectrophotometry and polarography. Tartrazine, Cochineal Red and Indigotin in amounts permitted by Polish legislation have been determined. PMID- 7481504 TI - [Use of the ICP-AES method for multi-element analysis of tobacco leaves]. AB - The method of the atomic emission spectrometry with an excitation in the inductively coupled plasma ICP-AES has been applied to the simultaneous analysis of the elements: Al, Cd, Cr, Cu, Fe, Li, Mg, Mn, Ni, P, Pb, S, Sr and V in the tobacco leaves samples CTA-OTL-1. The spectrometric analyses have been carried out on the emission spectrometer Spectroflame ICP M. About 300 mg of the sample and 1 cm3 of the concentrated HNO3 acid have been sealed in the quartz tube and mineralized by the pressure method. Since the ICP-AES technique is the most suitable to the direct analysis of the solutions, the main problem to solve was a mineralization of the sample. An application of the HNO3 acid alone did not give satisfactory results--unsolved residue remained. As a consequence the results for Mg, Ni, Pb were too low, whereas for the Al-nearly twice lower than the respective values given in the sample reference data. It has been observed that an addition of the conc. HF acid helps to dissolve the residue and, consequently, the optimalization of the mineralization process in the presence of the HNO3-HF mixture has been carried out. The results show that the appropriate amount of the HF acid to be added is 0.25 cm3. The other methods of mineralization using various mixtures of acids did not give good results. Only HNO3-HF mixture used in appropriate proportions give results in agreement with provided sample characteristics.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7481506 TI - [Method of standards preparation for colorimetric determination of ozone in the atmosphere]. AB - A new simple method of standards preparation for the colorimetric determination of "natural ozone" in the atmosphere in field conditions has been developed. An exactly known quantity of alcoholic iodine solution, adequate with respect to the stochiometric quantity of ozone is deposited on the iodine starch papers. In the comparison of the colours of these spots with the papers. In the comparison of the colours of these spots with the papers saturated with a mixture of drawing inks (burnt senna), a scale of constant standards was obtained for ozone determination in air by the iodometric method. The direct placing of a definite quantity of iodine on iodine starch papers enables the determination of small ozone concentrations (0.2 to 7 micrograms/m3) and brought about an essential increase in the accuracy and precision of the method. PMID- 7481501 TI - [Lead, cadmium, copper and zinc content in vegetables, gooseberry fruit and soil from gardening plots of Lublin]. AB - Lead, cadmium, copper and zinc contents in vegetables, fruits of gooseberries and in soil of Lublin gardening plots "Pionier" and "Podzamcze" both situated along heavy traffic streets, and "Pionier" additional to the close vicinity of automobile factory were determined by atomic absorption spectrophofometry. Statistical analysis of the results respected the streets or factory distances from the gardening plots, and the species of the vegetables. Levels of the determined elements in the most of the samples were lower than permitted by the Ministry of Health Regulation established in 1993. According to that Regulation a slight exceeding of zinc content in the red beetroots and in the overground parts of the leak were found. Lead content in the parsley root and in the dill was close to the tolerance limit. Both lead and cadmium of the soil of "Podzamcze" gardening plot significantly exceeded the levels considered as tolerable for unpolluted grounds, whereas the soil of "Pionier" contained lead close to limit of tolerance level. PMID- 7481507 TI - [Comparative analysis of two diffusion methods for radon Rn-222 estimation in atmospheric air by means of gamma ray spectrometry and liquid scintillation counting]. AB - The comparative measurements of radon Rn-222 concentration in indoor air of some buildings and in radon labour chamber have been conducted using two different diffusions methods. The results of parallel measurements using radon charcoal detectors and gamma-rays spectrometry with liquid alpha scintillation counting (Pico-Rad) have been presented. It was concluded that both methods offer the similar measurements possibilities of radon concentrations in the air from about 30 Bq/m3 to about 600 Bq/m3 with the real average divergence between both methods of about +/- 11%. PMID- 7481508 TI - A statistical evaluation of the occurrence of keratinolytic fungi in the sediments of two dam reservoirs. AB - Sediments from two dam reservoirs "Przeczyce" and "Plawniowice" were examined for keratinolytic fungi. The results show the dependence of keratinolytic fungi in sediments on the degree of water contamination with sewage. Ch. keratinophilum is species associated with sewage inputs to superficial water. PMID- 7481509 TI - [The influence of diesel fuel oil on the number of bacteria, fungi, actinomycetes and soil microbial biomass]. AB - In laboratory conditions the influence of Diesel fuel oil on the total number bacteria, fungi, actinomycetes and content of microbial biomass in soil was studied. The fuel oil was applied at dosage of 1, 2 and 5 g/100 g of soil. Samples of soil were analysed 1, 7, 14, 21 and 150 days after oil applied. Microbiological analyses involved determinations of the total number of bacteria and actinomycetes (on Bunta-Roviry medium) and fungi (on Martin's agar) by the plate method. The content of microbial biomass in soil was determined using the physiological method of Anderson and Domsch. Diesel fuel oil application caused the changes of total number of investigated groups of soil microorganisms. These results were confirmed by the investigated groups of soil microorganisms. These results were confirmed by the investigation on microbial biomass in soil. In the most of cases the fuel oil was the most stimulated for the number of bacteria and the most inhibited for actinomycetes in comparison with soil without oil (control). From soil with Diesel fuel oil addition Pseudomonas, Arhtrobacter, Bacillus and Penicillium, Aspergillus, Fusarium, Trichoderma strains were isolated. The possibility of the fuel oil as source of carbon utilization by these strains will be investigate. PMID- 7481510 TI - [Parents, from the opinions of students in the first classes of general education secondary schools]. AB - In cooperation with the workers of the Province Sanitary-Epidemiological Stations in Bydgoszcz, Gdansk, Koszalin, Cracow, Lodz, Poznan and Wroclaw an inquiry study was carried out of 839 students (517 girls and 322 boys) of the first classes of secondary schools, of whom 518 attended state schools and 321 attended schools run by non-governmental agencies. Since no statistically significant differences were found between girls and boys and between both types of schools, the results are presented for the whole studied population. Only 57% of the students regarded that in difficult situations they could rely on help and support of their parents; 55% considered that a mutual understanding existed between them and their parents; 7% described their relations with their parents as bad; 47 regarded their home atmosphere as calm and serene; 9% reported their families as contankerous; in 6% of the families quarrels between parents were frequent, and 15% of the students experienced such quarrels as particularly depressing, with 38% of girls and 14% of boys ruminating them even during lessons. Of interest were the data concerning spending of leisure time with parents. Time was spent with parents on school-days by 20% of the responders, and on Saturday and Sunday by 34%. On the other hand, time was not spent with parents on school-days by 37%, and during weekends by 16%. On weekdays 26% of female students from state-run schools and 12% of those from other schools spent the leisure time with their parents. Only 55% of the responding boys considered that the time spent with parents was sufficient. PMID- 7481505 TI - [Calcium, magnesium, copper and zinc in blood serum in men with femoral atherosclerosis]. AB - Calcium, magnesium, copper and zinc were determined in serum in men with femoral atherosclerosis. Also total cholesterol, HDL- and LDL-cholesterol were estimated. Subjects in control group were in the same range of age. The present study compares two methods of preparing serum for the flame atomic absorption spectrometry. In the first method (I), serum was diluted by the demineralized water. In the second method (II), serum protein was precipitated by trichloracetic acid (TCA). We found that the first method (I) was simpler and more precise than the other. The optimal proportions of serum and water amounted to 1 + 62 for calcium and magnesium and the coefficient of variation (C.V.) was 2.8%. For copper and zinc, the optimal dilution was 1 + 3 (C.V. = 1%). Total cholesterol was significantly higher (p < 0.05) and HDL-cholesterol was lower (p < 0.05) in subjects with femoral atherosclerosis as compared with controls. There was no difference in LDL-cholesterol. Subjects with femoral atherosclerosis had significantly higher level of all micro- and macroelements determined, then those of the control group (p < 0.05). However, higher serum copper does not necessary mean higher copper body status. It is possible that higher serum copper in femoral atherosclerosis reflects the transfer of copper from the tissue to the ceruloplasmin, as an acute phase reaction. On the other hand, the differences in serum magnesium, calcium, copper and zinc concentrations, may indicate the possible involvement of these elements in the disorder of total and HDL cholesterol in femoral atherosclerosis. PMID- 7481511 TI - [Standard radiography of the mandibular condyle in the dental office. The modified Parma and the Zimmer trans-orbital production technic using a retro alveolar radiologic unit]. AB - Trauma to the condylar area is rather frequent and the stomatologist should be capable of performing a standard radiographic exploration. Two incidences performed in the stomatologist's office using a dental tube appear to be essential. The modified Parma and the transorbitary Zimmer incidences are needed for lateral and anteroposterior views respectively. The equipment required for these views are described together with several examples. PMID- 7481512 TI - [The value of 3-dimensional computed tomography in the preoperative evaluation of mandibular condyle fractures]. AB - The recent expansion of surgical treatment for displaced condylar neck fractures should rely on a precise and careful preoperative radiological assessment. Two goals must be achieved. First the diagnosis must be confirmed by usually sufficient conventional imaging. Secondly, all the features of the fracture have to be analysed to come to the surgical decision and to choose the best osteosynthesis technique. Compared to traditional imaging, three dimensional computed tomography offers the possibility of a direct viewing. The extend of the fracture, the aspect and degree of displacement of the condylar fragment in relation to the ramus or the fossa are easily assessed. The examination and the integration of multiple axial or coronal bidimensional CT scan sections are no more necessary. The clearer perception of both the spacial relationships and delineation of these condylar fractures results in a better choice for surgical planning. PMID- 7481513 TI - [Primary carcinoma of the parotid gland. A retrospective study of 31 cases]. AB - The medical records of 31 patients with histologically proven primary carcinoma of the parotid gland seen between January 1980 and December 1992 were reviewed. The efficiency of combined treatment both on locoregional control and overall survival was assessed. Twenty nine patients underwent extensive surgery and postoperative radiotherapy, two patients underwent extensive surgery alone. Locoregional control of disease at the end of treatment was achieved in 29 patients (94%). Eleven patients had recurrences. For the complete series the overall two and five-year survival rates were 73% and 46% respectively. Even in case of locoregional control with combined treatment, overall survival rates are low because of distant metastases. PMID- 7481514 TI - [Our experience with parotid cysts]. AB - The cysts of the parotid gland are very rare lesions of congenital or acquired aetiopathogenesis. In most cases, the symptom is an unilateral painless swelling in the area of the parotid without any fixation to the overlying skin or any involvement of the facial nerve. These must be distinguished from neoplasms with a similar clinical presentation or neoplasms with a cystic component. Making the right preoperative diagnosis can be extremely difficult but the adequate therapy remains the superficial parotidectomy. However the final diagnosis is established histopathologically. The precise histological classification is often an elaborate task because of the similarities in the epithelial lining of the various cysts and the heterogeneity in their histogenic origin. In this study, the authors describe four cases of parotid cystis. They examine the clinical aspects, the preoperative diagnostic approach, the treatment and they analyse the histopathological findings. Finally, their results are compared with the data found in the international medical literature. PMID- 7481515 TI - [Palliative treatment of facial paralysis]. AB - Facial nerve palsy disrupts both the static and dynamic equilibrium of the half face involved. The imbalance worsens with age and senility further aggravates the situation. Palliative surgery can be used in complete intractable facial palsy or rarement for partial palsies or as a temporary treatment. The operation is proposed mostly for elderly patients and does not create other malformations or dysfunctions. Simple and effective reduction, cervicofacial lifting and plicature of the skin muscles is used with ocular protection (passive eyelid circle, lateral de-epidermalized skin flap, blepharorraphia) with or without use of locoregional tissues (orbito-naso-genial and labiogenial flaps) are usually sufficient. PMID- 7481516 TI - [A study model of the oral manifestations of HIV infection. The correlation and conformity of the WHO registry]. PMID- 7481517 TI - [Extemporaneous histologic examination of cutaneous carcinoma of the face]. AB - One hundred thirty-eight skin cancers of the face were reviewed retrospectively in order to evaluate the extemporaneous histology examination. The results obtained were satisfactory and in agreement with those in the literature. This technique is perfectly adapted to cancers with a facial localization. In our opinion, the indication for extemporaneous examinations should be widened to all cases of epithelioma of the face. PMID- 7481519 TI - [Sialometry of 156 healthy subjects. Physiologic factors which influence non stimulated saliva secretion]. AB - PURPOSE: Sialometria or measurement of salivary fluid is an important test for the patients with glandular disfunction. Biological parameters are being studied which may influence the results of the unstimulated salivary flow measurements. METHODS: Sialometria test, which was carried on 159 healthy subjects of a population (81 male and 79 females), studies habits (tobacco), age, sex, corporal weight, environmental factors, oral exploration (caries, lack of teeth and fillings) RESULTS: We found less salivary fluid in the elderly subjects (p = 0.0001), in women (p = 0.045) and with missing teeth (p = 0.0001). We did not find any variation respecting smoking habit, or corporal weight. CONCLUSIONS: Before carrying out the salivary resting flowmetry we ought to bear in mind the variations respecting sex, age and number of teeth missing when interpreting a resting flowmetry. PMID- 7481518 TI - [Bone lengthening: application to the mandible. Review of the literature]. AB - In literature, the authors demonstrate that progressive distraction osteogenesis, according to the Ilizarov method, is applicable to the mandible. Mandibular distraction is achievable through external approach or through oral approach. Further experimental and clinical studies will be necessary to define the most stable apparatus and the best indications that are currently hemifacial microsomia and facial asymmetry associated with T.M.J. ankylosis. PMID- 7481520 TI - Immune-mediated liver damage in chronic hepatitis C. AB - Although the exact mechanisms inducing the hepatic injury in chronic hepatitis C infection are still not completely elucidated, the evidence of a direct cytopathic viral effect is sparse. The overall majority of experimental studies and clinical observations among patients instead favour immune-mediated hepatocellular damage. This does not exclude the possibility of direct cytopathic viral effects partly contributing to the cell injury. The immune reaction is not always harmful, however. Instead, an intact immune system is probably important for virus clearance in acute infection and after interferon treatment. The therapeutic possibilities are still not adequate, and attempts to understand the immunologic events and manipulate them could perhaps in the future give us more effective tools in the management of this disease. PMID- 7481521 TI - Epidermal growth factor attenuates the sclerotherapy-induced biomechanical properties of the oesophagus. An experimental study in minipigs. AB - BACKGROUND: The effect of epidermal growth factor (EGF) on the biomechanical properties of the oesophagus subjected to sclerotherapy was studied in Goettingen minipigs by means of impedance planimetry. METHODS: Seventeen animals underwent three sessions of weekly endoscopic sclerotherapy. During these 3 weeks and for the subsequent 2 weeks they were treated with either EGF or placebo. After another 3 weeks an impedance planimetric study was done. Ten healthy non sclerosed pigs were studied as controls. Impedance planimetry was performed by stepwise pressure-induced balloon inflation for analysis of oesophageal cross sectional area 5 and 10 cm above the gastro-oesophageal junction (GEJ). RESULTS: Systemic treatment with EGF (total daily dose of 30 micrograms/kg/day, administered subcutaneously) for 3 to 7 days per week combined with a weekly paravenous injection of 20-40 micrograms/kg attenuated the oesophageal damage caused by sclerotherapy, implying less pronounced narrowing 5 cm above and less dilation 10 cm above the GEJ. CONCLUSION: These observations suggest a potential therapeutic role for EGF in attenuating sclerotherapy-induced oesophageal injury. PMID- 7481522 TI - Comparison of cholecystokinin, pentagastrin, and duodenal oleate in gastroprotection in rats. AB - BACKGROUND: Cholecystokinin (CCK) and gastrin show a potent influence on gastric secretion and motility, but their role in mucosal integrity has been little studied. METHODS: In this study the effects of CCK-8, pentagastrin, and duodenal oleate on acute gastric lesions induced by 100% ethanol were studied in rats. RESULTS: CCK-8 was about 13 times more potent than pentagastrin in protecting the gastric mucosa against ethanol damage. CCK released by duodenal oleate also protected gastric mucosa against this damage. The protective effects of CCK-8 were almost completely abolished by the blockage of CCK-A receptors with loxiglumide, whereas the protective effect of pentagastrin was completely abolished by L-365,260. The protective effects of CCK, pentagastrin, or duodenal oleate against ethanol injury were accompanied by a marked increase in luminal content of somatostatin, suggesting that this peptide is implicated in this protection. The protective activity of CCK and pentagastrin against ethanol injury was accompanied by a significant increase in gastric blood flow. Inhibition of nitric oxide (NO) synthase with NG-nitro-L-arginine methyl ester abolished almost completely both gastric protection and hyperemia induced by CCK and pentagastrin. Addition of L-arginine, but not D-arginine, restored the protective and hyperemic effects of CCK and pentagastrin. Pretreatment with the sulfhydryl blocking agent N-ethylmaleimide also abolished the protective and hyperemic effects of CCK and pentagastrin. The hyperemia, but not the protection, afforded by CCK and pentagastrin was reduced after sensory nerve deactivation with capsaicin. CONCLUSIONS: Both exogenous and endogenous CCK and pentagastrin exert protective activity against ethanol damage, and this effect is mediated through separate receptors, NO, and sulfhydryl-sensitive pathway. PMID- 7481523 TI - Low doses of pentagastrin stimulate gastric lipase secretion in man. AB - BACKGROUND: Gastric lipase is an important enzyme for dietary triglyceride digestion in normal subjects. Its regulation is unknown, as is the relation between the quantity and activity of the enzyme. METHODS: In a dose-response study we investigated the effect of low doses of pentagastrin (less than 1000 ng/kg/h) on the output of gastric lipase measured by a kinetic assay and an enzyme-linked immunosorbent assay (ELISA). RESULTS: In five healthy volunteers stepwise intravenous pentagastrin infusions of 0, 50, 100, 500, and 1000 ng/kg/h resulted in a stepwise increase in the lipase output, as measured with ELISA. However, the lipolytic activity, measured with a kinetic assay, decreased as the pH of the gastric contents decreased. CONCLUSION: We conclude that secretion of the gastric lipase is stimulated by pentagastrin, but the simultaneous secretion of acid counteracts the lipolytic activity of the enzyme when food is not present. PMID- 7481526 TI - Is Helicobacter pylori infection a real risk factor for gastric carcinoma? AB - BACKGROUND AND METHODS: That Helicobacter pylori has a role in the pathogenesis of gastric carcinoma is widely accepted, although not all doubts are definitively clarified. The purpose of this work was to detect the differences in presence and mean titer of anti-H. pylori antibodies between groups with gastric (n = 65), colonic (n = 70), and lung (n = 43) carcinoma. RESULTS: The highest prevalence of anti-H. pylori antibodies was found in patients with pulmonary carcinoma (88.4%), which significantly surpassed (p = 0.02) that in the group with gastric carcinoma (69.2%). The groups with colonic and gastric carcinomas failed to show any difference in this respect. Mean antibody titer was significantly higher in subjects with lung carcinoma than in those with gastric carcinoma (p = 0.005). This difference was unrelated to age. CONCLUSIONS: These results contradict the hypothesis assuming a relationship between H. pylori infection and the sequence of phenomena leading to gastric carcinoma. PMID- 7481524 TI - Analysis of antral biopsy specimens for evidence of acquired mucosa-associated lymphoid tissue in HIV1-seropositive and HIV1-negative patients. AB - BACKGROUND: There is conflicting evidence concerning the prevalence of Helicobacter pylori gastritis in HIV1-infected patients. Furthermore, a possible influence of immunodeficiency on the acquisition of mucosa-associated lymphoid tissue (MALT) in the antral mucosa remains to be elucidated. METHODS: Seventy seven consecutive HIV1-infected patients (mean age, 40.2 years) were compared in a prospective study with 77 HIV1-negative age-matched patients, using immunohistochemical stainings. RESULTS: In HIV1-infected patients the prevalence of H. pylori gastritis was lower and the inflammatory reaction less pronounced than in controls. Lymphoid follicles and intraepithelial B cells were significantly more often detected in HIV1-negative patients. CONCLUSIONS: Evidence of acquired MALT is only rarely found in HIV1-infected patients. These findings might contribute to the explanation of why low-grade gastric MALT lymphomas have not been reported in HIV1-infected patients so far. PMID- 7481527 TI - Absence of correlation between hepatic function and characteristics of migrating motor complexes in the gastrointestinal tract. AB - BACKGROUND: Cyclic changes in gallbladder filling and emptying during the migrating motor complex (MMC) cycle have been demonstrated by scintigraphy. However, a possible cyclic change in the hepatic function and handling of the pharmacologic agents used for scintigraphy during the MMC cycle could have an influence on these results. The aim of the present study was to investigate the hepatic handling of cholic acid and mebrofenin in relation to the MMCs of the gastrointestinal tract. METHODS: The plasma disappearance rate of 14C-cholic acid and the hepatic uptake and excretion of 99mTc-mebrofenin were examined during phase I and phase II of the MMC in six healthy male volunteers. RESULTS: The plasma disappearance rate of 14C-cholic acid showed a biexponential course with an initial rapid and late slow phase after a bolus injection. There were no significant differences between the initial or late plasma disappearance rate of 14C-cholic acid during phase I as compared with phase II. The results of the time activity curves from the 99mTc-mebrofenin scintigraphy showed an exponential rapid increase in radioactivity followed by an almost linear slow decrease after a bolus injection. There was no significant difference between phase I and phase II in any of the variables studied. CONCLUSION: The lack of a relationship between hepatic handling of cholic acid and mebrofenin and MMC excludes this as a possible source of error in the investigations of the dynamic function of the enterohepatic circulation and especially gallbladder motility by the use of either cholic acids or iminodiacetic acid derivatives as investigative agents. PMID- 7481528 TI - Gastric output of IgA in man: relation to migrating motility complexes and sham feeding. AB - BACKGROUND/AIMS: The immunologic reactivity of the gastric mucosa is poorly understood. The origin and dynamics of immunoglobulin A (IgA) occurring in the gastric lumen were investigated in healthy, Helicobacter pylori-negative volunteers. METHODS: Gastroduodenal manometric motility recordings were combined with gastric luminal perfusion, enabling calculation of gastric acid output and analysis of the total IgA output. RESULTS: Acid output and total IgA correlated with the migrating motility complexes (MMC). The gastric IgA release showed maximal values in association with gastric motility phase III (maximal motor activity) and lowest values during phases I and II (none or irregular motor activity). The IgA output correlated with neither swallowed saliva (as indicated by amylase in the gastric perfusate) nor duodenogastric reflux (as indicated by gastric occurrence of bilirubin and/or duodenally infused PEG4000). Stimulation of gastric acid secretion by sham feeding during phase II-like motor activity (n = 6) induced a rapid and transient doubling of IgA output. There was no significant correlation between gastric acid secretion and gastric IgA release. CONCLUSION: Substantial amounts of IgA are released into the human stomach, most likely originating from the gastric mucosa. The up-regulation of IgA release in association with the activity front of the MMC and anticipatory to food intake suggests a neuroendocrine control of gastric mucosal immune responses. PMID- 7481525 TI - Cluster analysis of Helicobacter pylori genomic DNA fingerprints suggests gastroduodenal disease-specific associations. AB - BACKGROUND: Helicobacter pylori infection is now accepted as the most common cause of chronic active gastritis and peptic ulcer disease. The etiologies of many infectious diseases have been attributed to specific or clonal strains of bacterial pathogens. Polymerase chain reaction (PCR) amplification of DNA between repetitive DNA sequences, REP elements (REP-PCR), has been utilized to generate DNA fingerprints to examine similarity among strains within a bacterial species. METHODS: Genomic DNA from H. pylori isolates obtained from 70 individuals (39 duodenal ulcers and 31 simple gastritis) was PCR-amplified using consensus probes to repetitive DNA elements. The H. pylori DNA fingerprints were analyzed for similarity and correlated with disease presentation using the NTSYS-pc computer program. RESULTS: Each H. pylori strain had a distinct DNA fingerprint except for two pairs. Single-colony DNA fingerprints of H. pylori from the same patient were identical, suggesting that each patient harbors a single strain. Computer assisted cluster analysis of the REP-PCR DNA fingerprints showed two large clusters of isolates, one associated with simple gastritis and the other with duodenal ulcer disease. CONCLUSIONS: Cluster analysis of REP-PCR DNA fingerprints of H. pylori strains suggests that duodenal ulcer isolates, as a group, are more similar to one another and different from gastritis isolates. These results suggest that disease-specific strains may exist. PMID- 7481530 TI - Enzymatic hydrolysis of long-chain alkanoylcholines in rat intestinal loops. AB - BACKGROUND: The hydrolysis of long-chain alkanoylcholines, presumably catalyzed by butyryl-cholinesterase (EC 3.1.1.8), in rat intestinal loops was studied. The substances have earlier been found to be rapidly degraded in vitro. METHODS: Radiolabeled substrates were used, and a radiochromatographic detection method was applied. RESULTS AND CONCLUSION: The long-chain alkanoylcholines were rapidly hydrolyzed. The rates of the reaction and the chain-length dependence were similar to those reported earlier in vitro. At high substrate concentrations the hydrolysis reaction was inhibited. This could be due to conformational changes of the enzyme, caused by the adsorption of the cationic amphiphile, or to a decrease in the free substrate concentration after incorporation of the amphiphilic ester into the lipid layer of the cell membranes. The enzymatic activity towards the substrates in different parts of the rat intestinal tract was also studied and found to be highest in the duodenum. PMID- 7481529 TI - Developmental, nutritional, and hormonal regulation of the pancreatitis associated protein I and III gene expression in the rat small intestine. AB - BACKGROUND: The pancreatitis-associated protein (PAP) I and III, but not PAP II, mRNAs are constitutively expressed in the small intestine of rats. METHODS: We studied expression of both PAP I and PAP III mRNAs during development and on nutritional and hormonal manipulations. RESULTS: Between day 20 of gestation and day 21 of age, PAP mRNAs could barely be detected. Their concentrations increased dramatically from day 21 to day 45 of age and remained constant thereafter. Rats adapted to a diet with low carbohydrate content showed a significant decrease in PAP mRNA concentrations. Finally, whereas thyroidectomy and ovariectomy induced a decrease in both mRNA concentrations, and adrenalectomy a limited decrease in PAP III mRNA only, diabetes and castration did not alter the expression of either gene. CONCLUSION: Gene expression of PAP I and III mRNAs is regulated in a coordinate manner in the rat small intestine during development and on nutritional and hormonal manipulations. PMID- 7481531 TI - Inability of Lactobacillus casei strain GG, L. acidophilus, and Bifidobacterium bifidum to degrade intestinal mucus glycoproteins. AB - BACKGROUND: Lactic acid bacteria have been suggested for use in the prevention of relapse of ulcerative colitis and of recurrent pouchitis. These strains may not damage the protective intestinal mucus glycoproteins. METHODS: Lactobacillus casei strain GG and strains isolated from a commercial fermented product (Lactobacillus acidophilus, Bifidobacterium bifidum, and a mesophylic lactic culture) were cultured in vitro on hog gastric mucin and human intestinal glycoproteins. Furthermore, germ-free rats were mono-associated with Lactobacillus GG and poly-associated with the other strains. Glycoproteins were isolated from rat distal ileum, cecum, and colon. Mucus degradation was established by assaying carbohydrates (hexosamines, hexoses, pentoses), proteins, and blood group antigenicity. RESULTS: All strains colonized the intestinal mucus but were not found in the deep crypts. Degradation of mucus glycoproteins was observed neither in vitro nor in vivo. CONCLUSION: The tested strains do not break down intestinal mucus glycoproteins and thus far are safe to use for therapy. PMID- 7481532 TI - Bacteriologic analysis of mucosal biopsy specimens for detecting small-intestinal bacterial overgrowth. AB - BACKGROUND: Although culture of luminal secretions is regarded as the most accurate diagnostic test for small-intestinal bacterial overgrowth, obtaining an aspirate is often difficult owing to the sparseness of luminal secretions present at the time of aspiration. Obtaining a mucosal biopsy specimen for bacteriologic analysis would overcome this problem. METHODS: Culture of small-intestinal and gastric aspirates and unwashed small-intestinal mucosal specimens was performed in 51 adult subjects investigated for small-intestinal overgrowth. RESULTS: Highly significant (r = 0.85-0.90; p < 0.0005) correlations were found between viable bacterial counts in small-intestinal luminal secretions and biopsy specimens. Small-intestinal bacterial overgrowth was present in 60.8% of subjects. When specimens weighing 4.0-84.0 mg were suspended in diluent, total aerobic and/or anaerobic bacterial counts > or = 10(2) CFU/ml were found to have 90.3% sensitivity and 100% specificity for small-intestinal bacterial overgrowth. CONCLUSION: Culture of an unwashed small-intestinal mucosal biopsy specimen is a useful alternative to culture of a small-intestinal aspirate for detecting subjects with small-intestinal bacterial overgrowth, especially when luminal secretions are scanty at the time of aspiration. PMID- 7481533 TI - A randomized surveillance study of patients with pedunculated and small sessile tubular and tubulovillous adenomas. The Funen Adenoma Follow-up Study. AB - BACKGROUND: We wanted to assess the influence of various surveillance intervals on the risk of new neoplasia after removal of pedunculated and small sessile tubular and tubulovillous adenomas. METHODS: After initial colonoscopic polypectomy patients were randomized to surveillance with either 2 years (group A) or 4 years (group B) between colorectal examinations. RESULTS: The cumulated risk of a patient having new adenomas was 35.0% (28.7-41.4%) in group A and 35.5% (28.4-42.7%) in group B after 48 months. The risk increased to 44.9% (36.0-53.9%) and 60.1% (48.5-71.7%), respectively, after 96 months. The risk of significant neoplasia (carcinoma or adenoma with villous structure, severe dysplasia, or diameter > 10 mm) was 5.2% (2.3-8.1%) and 8.6% (3.8-13.3%) after 48 months and 8.6% (4.2-13.0%) and 17.4% (7.6-27.2%) after 96 months. More than one adenoma at first examination was associated with higher risk of new adenomas. Furthermore, we found a tendency for age above 60 years and male gender to be associated with higher risk of new adenomas. More than two adenomas at first examination was the only factor found to be associated with a higher risk of new significant neoplasia. One patient in group A and two patients in group B developed cancer, which is not significantly different from the number expected (3.43) in the average Danish population (RR = 0.9, 0.2-2.6). CONCLUSION: After colonoscopy with removal of all polyps, colorectal examination at 4 years resulted in a similar risk of new adenomas compared with examinations at 2 and 4 years. However, new significant neoplasia tended to be more frequent when first surveillance was at 4 years. Extending the surveillance to 8 years also tended to increase the risk more in the group being examined every 4 years, but reduction of the number of surveillance examinations by more than 50% and a probable reduction of complications from surveillance examinations themselves may justify a recommendation for the longest interval. PMID- 7481534 TI - Anti-neutrophil cytoplasmic antibodies in chronic inflammatory bowel disease. Prevalence and diagnostic role. AB - BACKGROUND: Anti-neutrophil cytoplasmic antibodies (ANCA), originally found to be associated with vasculitis, have been reported to be present in chronic inflammatory bowel disease. Most often the ANCA staining pattern is of the perinuclear type (p-ANCA), although nuclear and cytoplasmic stainings are seen. Single studies have shown some of the antibodies to react with lactoferrin or cathepsin G; however, most studies have not been able to determine a main antigenic specificity. We studied the prevalence of ANCA in sera from 155 patients with ulcerative colitis, 128 patients with Crohn's disease, and 51 patients with coeliac disease. The presence of ANCA was correlated to disease activity, extent, and age of onset of the diseases. Furthermore, we tried to characterize the antigen specificity by enzyme-linked immunosorbent assay (ELISA), using elastase, lactoferrin, myeloperoxidase, proteinase 3, and cathepsin G as antigens. METHODS: The sera were screened for ANCA by indirect immunofluorescence. Anti-nuclear antibodies (ANA) were analysed on HEp2 cells, and ELISA for specific ANCA was performed using the antigens mentioned. RESULTS: Most of the sera with positive immunofluorescence had the p-ANCA type of pattern. Seventy-eight of 155 (50.3%) of the patients with ulcerative colitis were ANCA positive, compared with 31 of 128 (24.2%) of patients with Crohn's disease (p < 0.001). However, in the subgroup with Crohn's colitis, 16 of 44 (36.4%) were ANCA positive. Only 4 of 51 patients (7.7%) with coeliac disease showed positive immunofluorescence (p < 0.001 compared with ulcerative colitis). Less than 10% of the samples were positive in the specific ELISA assays; thus other than the most well known granule proteins can be the target for ANCA in ulcerative colitis. CONCLUSION: ANCA occur significantly more often in ulcerative colitis than in Crohn's disease. However, the prevalence of ANCA is rather high in Crohn's colitis. ANCA are thus of limited value in differentiating Crohn's colitis from ulcerative colitis. ANCA found in inflammatory bowel disease are different from those associated with vasculitis. The antigen(s) responsible remain to be determined. PMID- 7481535 TI - Disease activity courses in a regional cohort of Crohn's disease patients. AB - BACKGROUND: The study was a follow-up of an inception cohort of 373 patients with Crohn's disease. METHODS: Annual assessments, life table analyses, and Markov chain analyses, estimating the probability for remission and relapse with time, and working capacity were carried out. RESULTS: The clinical course of Crohn's disease differs markedly over time, from ever-relapsing cases to a quiescent course with remission for several years, interrupted by years with relapse. No predictive factors have been found for the subsequent course with regard to age, sex, extent of disease at diagnosis, and treatment in the year of diagnosis. The relapse rate within the year of diagnosis and the following 2 years, however, does correlate positively (p = 0.00001) with the relapse rate in the following 5 years. Furthermore, the relapse rate for 1 year during the disease course influences the relapse rate the following year, indicating a disease pattern over time with waves of at least 2 years' duration. A slight tendency towards burning out was found. The disease course reflected in working capacity for the patients showed that a minor part--up to 15% after 15 years--will become incapable and obtain disablement pension, while 75% of the patients each year are fully capable of work. Within 10 years 50% of the patients will not have experienced any year with impaired capacity for work. PMID- 7481537 TI - Role of nitric oxide in motility and secretion of the feline hepatobiliary tract. AB - BACKGROUND: Nitric oxide (NO) mediates inhibition of gastrointestinal smooth muscle cells via nonadrenergic, non-cholinergic (NANC) nervous pathways. The effect of NO on the absorption and secretion by the mucosa of the gastrointestinal and hepatobiliary tracts is less well known. The aim of this study was to evaluate the effects of a pharmacologic blockade of NO synthase on sphincter of Oddi activity, gallbladder function, and bile secretion and to demonstrate the presence of NO synthase-positive neurons in this region. METHODS: Experiments were conducted on anesthetized cats after blockage of noradrenergic and cholinergic neurotransmission. Flow resistance in the sphincter of Oddi, gallbladder fluid absorption and motility, bile outflow from the liver, and bile salt secretion were registered. RESULTS: Flow resistance exerted by the sphincter of Oddi increased dose-dependently in response to the NO synthase blocker NG nitro-L-arginine. The increase in flow resistance was reversed stereospecifically by L-arginine, the substrate for NO synthesis. No significant effects on bile secretion, gallbladder fluid transport, or gallbladder motility were observed. NO synthase-positive neurons were identified close to the sphincter of Oddi and in the gallbladder mucosa. CONCLUSIONS: This tonically active inhibitory NANC innervation of the sphincter of Oddi may be important in the physiologic regulation of the bile duct pressure. PMID- 7481538 TI - Scandinavian views on the non-surgical treatment of peptic ulcers, 1940-1975. Rationales for recommendations and results. PMID- 7481539 TI - Dynamic characteristic of gastro-oesophageal reflux in ambulatory patients with gastro-oesophageal reflux disease and normal control subjects. AB - BACKGROUND: The aim of the study was to investigate the dynamic characteristics of pathologic gastro-oesophageal reflux. METHODS: Five-channel ambulatory 24-h oesophageal pH monitoring was performed in 19 gastro-oesophageal reflux disease patients (age, 21-74 years) and in 19 healthy volunteers (age, 21-64 years). The pH was recorded at 3, 6, 9, 12, and 15 cm from the lower oesophageal sphincter (LOS), using a sample frequency of 4 Hz for each channel. Automated analysis included calculation of the ascending velocity of the refluxate and duration and extent (cm above the LOS) of all individual reflux episodes. RESULTS: In the patients more upright reflux episodes reached the proximal sensor than in the controls (20% and 11%, respectively, P < 0.01). The duration of the reflux episodes (measured at 3 cm above the LOS) was longer in the patients than in controls (P < 0.0001). This effect was independent of the proximal extent of the reflux episodes. Ascending velocities of upright acid reflux were higher in controls (1.8 to 2.7 cm/sec) than in patients (0.7 to 2.2 cm/sec; P = 0.01). CONCLUSIONS: The dynamic characteristics of pathologic reflux differ significantly from those of physiologic reflux. PMID- 7481540 TI - Cholecystokinin in the control of gastric acid secretion and gastrin release in response to a meal at low and high pH in healthy subjects and duodenal ulcer patients. AB - OBJECTIVE: In healthy subjects a gastric meal at low pH inhibits gastric acid secretion, possibly by reducing gastrin release, whereas duodenal ulcer (DU) patients have been reported to show a lack of this low pH inhibition of gastric secretion. METHODS: The intragastric pH profiles were measured in seven healthy subjects and seven DU patients after meals of pH 6.5 and 3.0 without or with pretreatment with loxiglumide (1.2 g orally), a selective antagonist of type-A cholecystokinin (CCK) receptors. During all tests (30 min before and 30, 60, and 90 min after each meal) plasma gastrin, CCK, and somatostatin were determined by specific radioimmunoassays. RESULTS: In healthy subjects a standard meal at pH 6.5 and 3.0 resulted in median 3-h intragastric pH of 3.8 and 2.8, respectively. In DU patients under the same conditions the pH 6.5 meal resulted in median 3-h intragastric pH of 3.4, and the acidified meal in pH 2.2. After pretreatment with loxiglumide the median pH after both meals was significantly lower in healthy controls but not in DU patients. After the pH 6.5 meal, in healthy subjects the plasma gastrin rose by 57%, CCK by 177%, and somatostatin by 39%, and in DU patients by 152%, 367%, and 125%, respectively. Pretreatment with loxiglumide led to a marked increase in plasma gastrin response to the pH 6.5 meal only in healthy controls and not in DU subjects, and it was accompanied by a significant increase in plasma CCK and a decrease in plasma somatostatin. The pH 3.0 meal resulted in a significantly smaller rise in plasma gastrin and a higher increase in CCK and somatostatin in both groups; again, after treatment with loxiglumide only healthy controls and not DU patients showed significant increase in plasma gastrin level. CONCLUSIONS: Acidification of meals results in the reduction of plasma gastrin and increase in plasma CCK and somatostatin in both healthy subjects and DU patients. DU patients differ from healthy subjects by virtually unchanged plasma gastrin response to a meal after CCK antagonism with loxiglumide, suggesting a defect in both gastric acid and gastrin inhibition by CCK in these patients. PMID- 7481536 TI - Transforming growth factor beta 1, extracellular matrix, and inflammatory cells in wound repair using a closed duodenal loop pancreatitis model rat. Immunohistochemical study. AB - BACKGROUND: Serial changes in the localization of various components of extracellular matrix in acute pancreatitis have been reported, but there have been no reports on serial changes in the localization of transforming growth factor beta and the determination of cells producing extracellular matrix. METHODS: In this study serial relationships between the localization of transforming growth factor beta 1, fibronectin and type-III collagen, inflammatory cells, and serum amylase levels in the process of tissue repair in acute pancreatitis were studied using a closed duodenal loop model rat. Furthermore, the cells producing transforming growth factor beta 1, fibronectin, and type-III collagen were investigated by immunoelectron microscopy. RESULTS: Three to 6 h after duodenal ligation slight localization of transforming growth factor beta 1 and fibronectin and inflammatory cell infiltration were observed in the interlobular space. Twelve to 24 h after duodenal ligation the infiltration of polymorphonuclear leukocytes and the deposition of transforming growth factor beta 1 and fibronectin were observed extensively in the interlobular and intralobular spaces. After release of the loop, infiltration of fibroblasts and marked deposition of fibronectin and type-III collagen were observed around the tubular complexes, but the deposition of transforming growth factor beta 1 was slight. Also, fibronectin and type-III collagen were shown to be produced by fibroblasts and acinar cells. CONCLUSIONS: These findings suggest that transforming growth factor beta 1 appears at the injured sites from the early stage of acute pancreatitis. Moreover, it is extensively related to the production of extracellular matrix such as fibronectin and type-III collagen. Furthermore, these substances are closely involved in the healing process of acute pancreatitis. PMID- 7481541 TI - Dyspeptic symptoms and gastric emptying of solids in patients with functional dyspepsia. Role of Helicobacter pylori infection. AB - BACKGROUND: Our aim was to investigate the relation between dyspeptic symptoms, gastric emptying of digestible and indigestible solids, and Helicobacter pylori infection in patients with functional dyspepsia. METHODS: We used isotopic labeling and radiologic techniques to study gastric emptying of a solid meal and of 10 radiopaque indigestible solids in 50 healthy volunteers and 50 patients with functional dyspepsia. In addition, we determined the presence of seven symptoms of dyspepsia and added the score for each symptom to obtain an index of dyspepsia for each patient. RESULTS: Seventy-eight per cent of our dyspeptic patients had gastroparesis to a solid meal, and 68% to indigestible solids. We found no apparent relation between gastroparesis or H. pylori infection and dyspeptic symptoms separately or as an index of dyspepsia. Moreover, the presence of the bacteria was not related to gastroparesis to a solid meal or to indigestible solids. CONCLUSIONS: We conclude that neither symptoms of dyspepsia nor H. pylori appears to be related to gastroparesis to solids. H. pylori infection is not related to dyspeptic symptoms. PMID- 7481542 TI - Helicobacter pylori seropositivity among Swedish adults with and without abdominal symptoms. A population-based epidemiologic study. AB - BACKGROUND: The role of Helicobacter pylori in functional dyspepsia is unclear. The aim of this population-based study was to determine whether the prevalence of H. pylori infection is higher among people with dyspepsia or irritable bowel syndrome (IBS) than among symptomless persons after control for age, sex, and socioeconomic status. METHODS: In a postal questionnaire we asked a representative sample (20-79 years; n = 1260) from a Swedish municipality about abdominal symptoms in the preceding 3 months. A randomly selected subsample, 50 with dyspepsia, 50 with IBS, and 50 symptomless, matched with regard to age, sex, and education, were tested for the presence of IgG antibodies to H. pylori, using the HM-CAP immunoassay. RESULTS: Fifty-five persons (38%) were H. pylori seropositive. The seroprevalence among dyspeptics (33%) did not exceed that in healthy people (48%) or in those reporting IBS (33%). The prevalence increased with age and with lower social class, but the latter association disappeared when age was taken into account. Neither sex nor symptom intensity predicted Helicobacter seropositivity. CONCLUSION: Our data are incompatible with an important aetiologic role for H. pylori in functional dyspepsia. PMID- 7481543 TI - In vitro toxicity of gluten peptides in coeliac disease assessed by organ culture. AB - BACKGROUND: We have used organ culture to investigate the in vitro toxicity of three oligopeptides corresponding to amino acids 31-49 (peptide A), 202-220 (peptide B), and 3-21 (peptide C) of A-gliadin, Frazer's fraction III (FFIII), and ovalbumin. METHODS: Eight to 14 jejunal biopsy specimens were obtained from each of 8 treated and 7 untreated coeliac patients and 5 normal controls and cultured for 18 h in organ culture with test peptide (1 mg/ml) or medium alone. Mean enterocyte cell heights (ECH) were compared with paired values for specimens grown in medium alone. RESULTS: A significant reduction in the mean of the ECH values for each of the patient groups was observed with peptide A and FFIII in both treated (p = 0.01 and 0.02, respectively) and untreated (p = 0.03 and 0.01) coeliac patients when compared with tissue incubated with medium alone. No significant changes in the mean ECH value were noted in any of the patient groups in tissue incubated with peptide B, peptide C, or ovalbumin as compared with those with medium alone. CONCLUSIONS: These results suggest that peptide A is toxic in vitro to the jejunal mucosa of both treated and untreated coeliac patients, correlating with recent findings that this peptide exacerbates coeliac disease in vivo. PMID- 7481544 TI - Gut failure in pediatric and adult patients. Candidates for small-bowel transplantation in southern Finland. AB - BACKGROUND: This study investigated the need for small-bowel transplantation (SBT) in southern Finland. In addition, we evaluated the effect of colon resection on the time required for intestinal adaptation in patients with gut failure. METHODS: Mortality, morbidity, intestinal adaptation time, and surgical management were evaluated retrospectively in 24 patients (12 adults and 12 children) with jejunoileal exculsion. Of the 24 patients 19 had undergone accompanying colon resection. RESULTS: Intestinal adaptation had occurred in 12 of 18 survivors (67%). Two unadapted survivors were considered to be currently suitable candidates for SBT. The length of the remaining colon was inversely correlated (correlation coefficient r = -0.789; p < 0.0001) with the duration of intestinal adaptation. CONCLUSIONS: Each year one to two patients per 1.27 million people could benefit from SBT in southern Finland. The length of the remaining colon is a major determinant of the duration of intestinal adaptation in patients with massive jejunoileal resection. PMID- 7481545 TI - Clinical and biologic features of adenomatosis coli in Northern Italy. AB - BACKGROUND: Familial adenomatous polyposis (FAP) is a hereditary disease characterized by more than 100 adenomas scattered in the large bowel and by various extracolonic manifestations. We proposed a) to establish the frequency of the disorder in Northern Italy, b) to describe the most relevant clinical findings, and c) in a subgroup of 21 patients (from 8 families), to evaluate the spectrum of mutations of the APC gene. METHODS AND RESULTS: Patients with FAP diagnosed between 1961 and 1991 were referred to our Study Group from surgery and gastroenterology units of the region Emilia-Romagna. The incidence of FAP was in the order of 1 in 16,500, with about a third of patients being 'single' cases. Colorectal malignancies were present in 75.6% of symptomatic patients but absent in most (93.75%) of the asymptomatic family members ('call-up' individuals). Gastric, duodenal, and jejunal adenomas were found in 8.2%, 30.6% and 53.3% of the investigated patients, respectively. Congenital hypertrophy of the retinal pigment epithelium and occult jaw lesions were seen in 64.7% and 39.5% of FAP patients but only in 0.5% and 2.5% of a matched, by age and sex, control population. These two clinical markers had a specificity of 99% and 97%, although their sensitivity was 64% and 39%. Finally, mutations of the APC gene were detected in 6 families (16 affected individuals) of the 8 families (21 affected individuals) tested; no correlation could be found between genotype and phenotype. CONCLUSIONS: This study confirms that early diagnosis is essential for an appropriate management of FAP patients, although this aim remains elusive in single cases. High-risk individuals are ideal candidates for APC gene mutation analysis, which should be offered to all first-degree relatives of affected patients. PMID- 7481546 TI - Intestinal permeability to polyethylene glycols in monozygotic twins with Crohn's disease. AB - BACKGROUND: A deranged mucosal permeability, demonstrated in several studies, has been proposed to play a role in the pathogenesis of Crohn's disease. The possibility of a genetically determined alteration of paracellular transport has been indicated in some investigations. The identification of a group of monozygotic twin pairs concordant and discordant for Crohn's disease, prompted this investigation. METHODS: Intestinal absorption after an oral load of different-sized polyethylene glycols (mol.wt, 458-810) was studied as 6-h urinary recovery. The study groups comprised twins with Crohn's disease (n = 19) and their healthy twin siblings (n = 9), non-twin patients with Crohn's disease (n = 14), and healthy controls (n = 30). RESULTS: No differences were found in the absorption of polyethylene glycols between the study groups. CONCLUSION: The results give no support to the hypothesis of a genetically determined intestinal leakiness in Crohn's disease. PMID- 7481547 TI - Serum nitrate levels in ulcerative colitis and Crohn's disease. AB - BACKGROUND: Nitric oxide is an important mediator in inflammatory and autoimmune mediated tissue destruction and may be of pathophysiologic importance in inflammatory bowel disease. We studied whether serum levels of nitrate, the stable end-product of nitric oxide, are increased in active Crohn's disease or ulcerative colitis, in comparison with quiescent disease and healthy controls. The setting was the gastroenterology unit of the Free University Hospital, Amsterdam. METHODS: In 146 patients--75 with ulcerative colitis and 71 with Crohn's disease--and 33 controls serum nitrate was measured by the Griess reaction after enzymatic conversion of nitrate to nitrite with nitrate reductase. RESULTS: Median serum nitrate concentrations did not differ statistically significantly between ulcerative colitis (median, 34.2 mumol/l; range, 15.6-229.4 mumol/l), Crohn's disease (median 32.3 mumol; range 13.2-143.2 mumol/l), and healthy controls (median, 28.7 mumol/l; range, 13.0-108.4 mumol/l). However, when active ulcerative colitis patients (median, 44 mumol/l; range, 29.1-229.4 mumol/l were compared with inactive ulcerative colitis patients (median, 31.2 mumol/l; range, 15.6-59.7 mumol/l), a significant difference in nitrate concentration was found (p < 0.0001). A significant positive correlation was found between serum nitrate levels in ulcerative colitis and erythrocyte sedimentation rate (ESR) (r = 0.30, p - 0.01), leucocyte count (r = 0.27, p = 0.02), and thrombocyte count (r = 0.24, p = 0.04). Comparing active Crohn's disease patients (median, 37.5 mumol/l; range, 13.2-143.2 mumol/l) with inactive Crohn's disease patients (median, 31.3 mumol/l; range, 14.5-92.3 mumol/l) also showed a significant difference in serum nitrate concentration (p < 0.009). Serum nitrate levels correlated with the ESR (r = 0.26, p = 0.028) and serum albumin (r = 0.38, p = 0.004) as well. CONCLUSION: Nitric oxide production is increased in both active ulcerative colitis and Crohn's disease and may be implicated in the pathogenesis of inflammatory bowel disease. PMID- 7481548 TI - Interferon-alpha 2b, with or without prior hepatic artery embolization: clinical response and survival in mid-gut carcinoid patients. The Norwegian carcinoid study. AB - BACKGROUND: Mid-gut carcinoid tumours often present with liver metastases, and survival has then been less than 2 years in earlier reports. We have evaluated the effects of interferon therapy on clinical response and survival, with or without hepatic artery embolization in these patients. METHODS: In a prospective study 30 female and 12 male patients, aged 23 to 75 years, with mid-gut carcinoid tumours and liver metastases underwent surgery with removal of as much as possible of their primary tumour. If technically feasible, embolization of hepatic arteries was performed in the absence of contraindications. Seventeen patients were embolized, and all patients received interferon-alpha 2b treatment for 1 year. Response factors were computer tomography (CT) measurement of the largest liver metastasis and the 24-h urinary excretion of 5-hydroxyindoleacetic acid (5-HIAA). After 12 months patients with objective response or stable disease either continued or withdrew from interferon therapy. Survival was estimated when all patients had been observed for at least 36 months. RESULTS: Nine patients reduced the dose, and five withdrew from interferon treatment owing to side effects the 1st year. Three patients died. Fifteen patients (39%) showed objective response 12 months after inclusion. Cumulative 5-year survival estimated from inclusion was 37.5% in all 42 patients but 71.4% in those who continued interferon therapy. The difference in survival between the interferon treated and those who withdrew from interferon therapy at 12 months was significant when embolization was corrected for in a Cox model (p < 0.0125). The seemingly increased survival in embolized versus non-embolized patients did not reach statistical significance (p = 0.07). CONCLUSION: Interferon induced an objective response in mid-gut carcinoid patients as judged by the 24-h urinary 5 HIAA excretion. Patients receiving continuous interferon therapy showed improved response and survival compared with patients who stopped the treatment. Regardless of medical therapy, more survivors and more responders, as evaluated from CT measurements, were found among the embolized patients than among the non embolized. Embolization could, however, not be shown to have a significant effect on survival. PMID- 7481549 TI - Endoscopic manometry of the sphincter of Oddi: its usefulness for the diagnosis and treatment of benign papillary stenosis. AB - BACKGROUND: Endoscopic manometry is considered useful to identify dysfunction of the sphincter of Oddi (SO) and to predict in which patients good results can be expected after endoscopic sphincterotomy, but this has not been definitively demonstrated. METHODS: Endoscopic manometry of the SO was used in a group of 30 patients with benign papillary stenosis (BPS), in comparison with 30 control subjects. During endoscopic manometry an intravenous bolus of cholecystokinin octapeptide was given to 12 patients and to 10 controls. In 24 BPS patients endoscopic sphincterotomy was performed. RESULTS: No significant differences were observed between controls and patients with regard to median values of SO basal (20 and 21.5 mmHg) and peak pressure (123 and 126 mmHg), wave amplitude (100 mmHg), frequency (4 waves/min), and propagation of the common bile duct/duodenum gradient (12.5 and 12.1 mmHg). In two BPS patients a paradoxic response to CCK-OP was observed. Endoscopic sphincterotomy, performed in 24 BPS patients (17 with SO basal pressure less than 40 mmHg and 7 with more than 40 mmHg), gave good results in 23, without any complication. No differences were observed in the results of the endoscopic sphincterotomy among patients with basal pressure more than 40 mmHg and those with less than 40 mmHg. CONCLUSIONS: On the basis of this study, manometric data do not seem helpful for diagnosis of BPS or to discriminate which patients can be treated with endoscopic sphincterotomy. PMID- 7481550 TI - Lymphangioma of Vater's ampulla: a rare cause of obstructive jaundice. Endoscopic therapy. AB - BACKGROUND: We report endoscopic excision of a lymphangioma in an infrequent location, Vater's ampulla, with resolution of the obstructive jaundice caused by this tumor. METHODS: An endoscopic papillotomy was performed together with polypectomy of the tumoral mass. RESULTS: The patient evolved satisfactorily, with jaundice and biochemical cholestasis disappearing. CONCLUSIONS: We believe endoscopic therapy is the ideal treatment for small lymphangiomas in the digestive tract. PMID- 7481553 TI - Nitric oxide synthase pathway may mediate human natural killer cell cytotoxicity. AB - The present study provides evidence that the human natural killer (NK) cell effector mechanism causing target cytolysis has a requirement for L-arginine. In a deficient medium (DM) containing only salts, buffer system and glucose, NK cell mediated cytotoxicity was found to decrease by 70% as compared to that obtained in a complete medium (CM). However, adding L-arginine to such DM could restore the activity of NK cells to the normal level. Many other components of CM, such as serum, glutamine and vitamins did not improve NK cell-mediated killing in DM. When all amino acids except L-arginine were added to DM only a partial recovery of NK cell functional cytolysis was seen. L-arginine enhanced the NK cell activity in a dose-dependent manner. Additionally, the inhibitor of both inducible and constitutive nitric oxide synthase, N-monomethyl-L-arginine (L NMMA) inhibited NK cytolytic activity in DM supplemented with L-arginine indicating participation of nitric oxide (NO). The results also show that the stimulatory effect of L-arginine on human NK cell-mediated cytotoxicity was accompanied by an increase in NO formation as determined by accumulation of nitrite and citrulline. L-NMMA gave a dose-dependent reduction in NO generation as well. The nitrite and citrulline production dose-dependently correlated with not only the concentration of L-arginine in the cultivation medium, but also the enhanced NK cell-mediated cytolysis. Taken together, these findings could define a L-arginine/NO-linked effector mechanism in human NK cells. Nitrite and citrulline were not formed when NK cell-mediated target cell killing took place in a L-arginine-free DM supplemented with additives. Thus, it appears as if human NK cells may cause target cell killing via both NO-dependent and -independent processes. PMID- 7481552 TI - Hyperoncotic dextran and systemic aprotinin in necrotizing rodent pancreatitis. AB - BACKGROUND: Dextrans improve pancreatic microcirculation in acute experimental pancreatitis. They could therefore facilitate the transport of a protease inhibitor to ischemic areas of tissue injury and be of additional benefit. METHODS: To compare the effects of dextrans with and without aprotinin, necrotizing pancreatitis was induced in 33 male dextran-resistant Wistar rats by intraductal infusion of low-dose glycodeoxycholic acid (10 mmol/l) followed by intravenous cerulein (5 micrograms/kg/h) for 6 h. Three and four hours after the start of the cerulein infusion the animals received infusions of either Ringer's lactate (RL) (12 ml/kg), 70,000 Da dextran (10%) (DEX-70) (4 ml/kg) alone, or DEX 70 (4 ml/kg) with aprotinin (5000 IU/kg) (DEX-70/A). RESULTS: The death rate was 60% within 9 h in the RL group (6 of 10) but only 10% in the DEX-70 group (1 of 10) (p < 0.03; Fisher's exact test) and 23% in the DEX-70/A group (3 of 13). Histomorphometry demonstrated a significant reduction of acinar necrosis in both treatment groups compared with control animals (p < 0.014; t test). Total amounts of trypsinogen activation peptides (TAP) in ascites were also significantly lower in these groups (p < 0.05; t test). CONCLUSIONS: DEX-70 given 3 h and 4 h after induction of pancreatitis significantly reduced the levels of TAP, limited acinar necrosis, and improved survival rate in acute necrotizing rodent pancreatitis. There was no additional benefit from the combination with aprotinin. PMID- 7481551 TI - Serum levels of soluble interleukin-2 receptor in chronic hepatitis C treated with interferon-alpha. AB - BACKGROUND: Serum levels of soluble interleukin-2 receptor (sIL-2R) seem to serve as a marker for the activation of T lymphocytes. The aim of this study was to evaluate the clinical significance of such levels in patients with chronic hepatitis C (CHC) treated with interferon. METHODS: We measured serum levels of sIL-2R in 37 patients with CHC before and after treatment with recombinant interferon-alpha. Serum receptor levels were then compared with the response of the hepatitis C virus (HCV)-RNA level in serum after interferon. RESULTS: Receptor levels were significantly higher in the patients with chronic persistent hepatitis and chronic active hepatitis than in normal controls (p < 0.01). There was a weak correlation between serum sIL-2R and alanine aminotransferase (ALAT) levels (r = 0.14, p = 0.010). Patients were then classified into three groups on the basis of the effect of interferon treatment on HCV-RNA levels in serum: sustained response (SR; n = 21), non-sustained response (NSR; n = 14), and nonresponse (NR; n = 2). Before and during interferon treatment the serum sIL-2R level remained increased in the SR group and in the combined groups with NSR or NR. However, after interferon was withdrawn, the serum sIL-2R decreased in the SR group but remained significantly increased in the combined response group (p < 0.01-0.05). CONCLUSION: This finding seems to reflect the disappearance of HCV RNA from the serum of the patients with an SR, and monitoring of sIL-2R levels may therefore be of value as an adjunct to the measurement of serum ALAT and HCV RNA in evaluating the response to the interferon therapy for CHC. PMID- 7481554 TI - Down-regulation of CD59 (protectin) expression on human colorectal adenocarcinoma cell lines by levamisole. AB - The vulnerability of tumour cells to complement-mediated immune attack is regulated by membrane associated molecules. Recently, we have shown that the expression of the membrane attack complex inhibitor CD59 is enhanced on colonic adenocarcinoma cells compared to normal colonic epithelial cells. CD59 was shown, in the same study, to protect the tumour cells from complement-mediated lysis. Levamisole (LMS), used in conjunction with 5-fluorouracil as adjuvant therapy, reduces the incidence of colon cancer relapse following surgical resection. This led to our investigation of the effect of LMS on CD59 expression and function on the human colorectal cell lines HT29 and Caco-2. When cultured in the presence of 10 microM LMS, the cells reduced their expression of CD59 in a time-dependent manner. LMS treated HT29 cells were more sensitive to lysis by complement than control cells, and the reduction in CD59 expression was shown to be partly responsible for this. A reduction in CD59 expression will augment complement mediated immune surveillance and may contribute to LMSs anti-tumour activity in vivo. PMID- 7481556 TI - Characterization of monoclonal antibodies to the alpha IIb beta 3 integrin on bovine platelets. AB - A set of monoclonal antibodies (MoAbs) to leucocyte antigens is an essential tool to identify different cell types and functional membrane molecules involved in immune responses. Since no MoAbs existed to bovine integrins, except against the beta 2 subfamily, we generated MoAbs to beta 3 integrin after the immunization of mice with bovine platelets. Two MoAbs, IL-A164 (IgG2a) and IL-A166 (IgG1), were selected that reacted specifically with bovine platelets and detected the same membrane molecule. The antigen was a heterodimer of two polypeptide chains of 122 kDa and 95 kDa as resolved by SDS-PAGE under reducing conditions. Although the Mr of the smaller subunit is identical to that of beta 2 integrin, preabsorption with an antibody to beta 2 (or CD18) did not remove the bovine antigen. Comparing the molecular masses of the two subunits in reduced and non-reduced forms showed a pattern that was similar to that of human GPIIb/IIIa (also called alpha IIb beta 3 or CD41a). Reduction of the bovine molecule increased the apparent Mr of the light chain from 76 kDa to 95 kDa, while the heavy subunit changed from 136 kDa to 122 kDa. As with human GPIIb, the decrease in Mr of the alpha-subunit is probably a result of a small disulphide-linked polypeptide, although no additional evidence for this was detected for the bovine integrin. Sequencing of the N-terminal amino acids of both bovine polypeptides showed identity of the bovine integrin with human GPIIb/IIIa. PMID- 7481555 TI - Antigen-pulsed, interleukin-4-treated B cells activate primed T cells in vitro but not naive T cells in vivo. AB - The ability of B cells to act as effective antigen-presenting cells is a source of debate which centres on the degree of activation of either B cells or T cells. We have investigated whether B cells treated with interleukin 4 (IL-4) can express the two signals required to activate T cells: MHC Class 2/antigenic peptide complexes(signal 1) and the costimulatory molecules B7-1 and B7-2 (signal 2). We have also determined whether these cells could activate antigen experienced T cells in vitro and whether they could prime naive T cells in vivo. We found that B cells expressed abundant MHC Class 2 molecules and moderate levels of B7-2 after 24 h culture in IL-4 with or without purified protein derivative (PPD) but B7-1 was not detectable. PPD-pulsed, IL-4 treated B cells induced antigen-experienced T cells to proliferate in vitro but these cells failed to prime naive T cells in vivo when injected into mice. We conclude that signals, in addition to those induced with IL-4, are required for B cells to initiate an immune response to antigen. PMID- 7481557 TI - Short-term administration of selected anti-T-cell receptor V beta chain specific MoAb reduces sialadenitis in MRL/lpr mice. AB - Sialadenitis develops spontaneously in MRL/Mp mice bearing a lymphoproliferative gene, lpr (MRL/Mp-lpr/lpr). Based on recent observations of an oligoclonal expansion of T-cell receptor (TCR) expressing V beta chain families (V beta 4, V beta 8.1,2, V beta 10b) in salivary glands of these mice we have initiated selective antibody therapy. Treatment with monoclonal antibodies (MoAb) specific for T cells expressing a mixture of TCR V beta 4, V beta 8.1,2 and V beta 10b was applied to MRL/lpr mice before and after the spontaneous development of sialadenitis. The in vivo treatment with V beta 4, V beta 8.1,2 and V beta 10b MoAb did not prevent the development of sialadenitis. However, in animals with established sialadenitis, treatment with the MoAb significantly decreased the inflammation compared with the control groups. Immunohistochemical staining of cell phenotypes demonstrated a change in the ratio of CD4/CD8 in the animals with established sialadenitis. Altogether, these findings illustrate that it is possible to modulate sialadenitis and infiltrate cell phenotypes in vivo in MRL/lpr mice with specific anti-TCR V beta MoAb treatment. PMID- 7481558 TI - A single major transcript encodes the membrane-bound form of rat immunoglobulin E. AB - The primary structure of the membrane-bound form of rat immunoglobulin E was determined by PCR amplification and nucleotide sequence analysis of its mRNA. The sequence was found to correspond to the previously identified membrane exons of the rat epsilon chain gene. The donor splice site in the C4 exon was mapped to a position 35 nt upstream of the stop codon for the secreted form of rat IgE. Therefore, the membrane-bound IgE lacks the 12 C-terminal amino acids present in the secreted form of the protein. Recently, five novel epsilon chain transcripts were isolated from human IgE producing B-cells or B-cell lines. Four of these transcripts encode proteins which differ in their C-terminal ends from the classical membrane or secreted forms of human IgE. To investigate if these transcripts were likely to represent functional mRNAs, their evolutionary conservation was studied by screening a rat IgE producing B-cell line for the expression of similar transcripts. By PCR amplification and cloning of transcripts, containing both the C3 and the M2 exons, approximately 10,000 independent cDNA clones were obtained. These clones were screened with probes directed against regions specific for each of the five novel human epsilon chain mRNAs. However, no evidence was found for the presence of transcripts with a similar structure, indicating that no specific function associated with these transcripts and their corresponding proteins has been conserved between human and rat. The lack of additional M2-containing transcripts in the rat suggest that the novel human IgE transcripts are byproducts of differential splicing and that they most likely encode proteins with no evolutionarily important function. PMID- 7481559 TI - Immunobiological studies on experimental visceral leishmaniasis. IV. Kinetics of evolution of disease-promoting versus host-protective cells of monocyte macrophage lineage and their characterization. AB - The evolution of cells of the monocyte-macrophage lineage (MML cells) in the spleen of Leishmania donovani (LD) infected BALB/c mice was studied. Spleen cells were fractionated on a discontinuous percoll gradient and adherent cells (AC) were purified from fractionated spleen cells by adherence steps that appeared at the interfaces of 25-35%, 35-40%, 40-45% and 45-50% percoll gradients. The AC were characterized as MML cells on the basis of positive staining for non specific esterase. Adherent cells that appeared at the interfaces of 25-35% and 40-45% were defined as A and C, respectively, and both of them showed extreme variation in a progressive infection. It was observed that A supported parasite replication whereas C remained refractory when infected with LD in vitro. Furthermore, when A cells and C cells were used as antigen-presenting cells to stimulate mixed population of IFN-gamma producing and IL-4 producing T-cells, it was observed that IL-4 and IFN-gamma were the predominating cytokine in the T cell supernatant, respectively. Both A and C were found to be increased hand-in hand up to 5 months of infection and from then on A decreased and C increased in their numerical strength (A-C reciprocity). The evolution of A-C reciprocity coincided with the gradual reduction in the parasitaemia in the spleen suggesting that this may contribute to the acquisition of anti-leishmania immunity. PMID- 7481560 TI - Lack of functional similarity between complement factor H and anticardiolipin cofactor, beta 2-glycoprotein I (apolipoprotein H). AB - Beta 2-glycoprotein I (beta 2-GPI) is a 50 kDa protein in human plasma composed of five repeating complement control protein modules thereby closely resembling complement factor H which has 20 such units. Both beta 2-GPI and factor H (150 kDa) have binding sites for negatively charged polyions. beta 2-GPI has been shown to act as a cofactor for antiphospholipid antibodies upon their binding to anionic phospholipids. In factor H the polyanion recognition site participates in the discrimination between alternative pathway activating and non-activating surfaces. In light of the structural similarity between beta 2-GPI and factor H we have examined whether beta 2-GPI has a role in the alternative complement pathway recognition process. Both activators (zymosan) and non-activators (sheep erythrocytes) of the alternative complement pathway were coated with C3b. Radiolabelled factor H was observed to recognize C3b on both surfaces, whereas beta 2-GPI bound to neither. In competition experiments beta 2-GPI could not prevent the association of 125I-H with either non-activator or activator bound C3b. Conversely, factor H could not replace beta 2-GPI as a cofactor for antiphospholipid antibodies upon their binding to anionic phospholipids. It is concluded that beta 2-GPI and factor H, despite similarities in structure, exhibit distinct, non-overlapping functions. PMID- 7481561 TI - Calprotectin-mediated zinc chelation as a biostatic mechanism in host defence. AB - The S-100 Ca2+ binding protein, calprotectin, isolated from neutrophil lysates, has been reported to exhibit zinc reversible biostatic activity in vitro. We verified these findings with C. albicans and investigated whether the growth inhibition resulted from zinc deprivation due to chelation by calprotectin. Calprotectin concentrations of 250 micrograms/ml significantly inhibited the growth of C. albicans. This was reversed by supplementing culture medium with 10 microM ZnSO4. Incubation of calprotectin in culture medium for 24 h prior to inoculation significantly reduced the minimum inhibitory concentration. When this latter medium was ultrafiltered to remove the calprotectin and then inoculated with C. albicans, significant growth inhibition was still present: again it was reversed by zinc. These findings implicate zinc chelation as a novel, potentially important host defence function of an abundant neutrophil protein. PMID- 7481562 TI - Both immunization with protein and recombinant vaccinia virus can stimulate CTL specific for the E7 protein of human papilloma virus 16 in H-2d mice. AB - The transforming protein E7 of human papilloma virus type 16 can stimulate cytotoxic T lymphocytes (CTL) which can protect experimental animals against growth of E7 expressing tumour cells. In this study we compared CTL responses in mice immunized with either E7 protein in MF59 adjuvant or with recombinant vaccinia virus expressing E7 (Vac-E7). We have chosen H-2d mice because no E7 specific CTL responses have been described in this MHC haplotype. Immunization of these mice with Vac-E7 generated CTL which lysed target cells infected with Vac E7 or transfected with the E7 gene. CTL from mice immunized with E7 protein in MF59 adjuvant showed specificity for the same target cells. Antibody blocking experiments revealed that both immunization with Vac-E7 and E7 protein stimulated CD8+ effector CTL. The find specificity of CTL induced by the two immunization protocols was similar. A major CTL epitope was mapped to the carboxyl terminal amino acids 48-98 of the E7 protein. Peptide isolation from E7 expressing cells followed by HPLC separation indicated that CTL induced by immunization with protein and Vac-E7 recognized the same HPLC purified peptide fractions. Together, the study suggests that vaccines based on protein can activate CTL with similar fine specificity to CTL induced by vaccines based on recombinant vaccinia virus. PMID- 7481563 TI - Distributions of HLA class II alleles in autoantibody subsets among Norwegian patients with systemic lupus erythematosus. AB - In order to find potential correlations between HLA class II alleles and anti-SS A, -SS-B, -Sm and anti-snRNP responses among Norwegian patients with systemic lupus erythematosus (SLE), HLA-DRB1, -DRB3*0101, -DQA1 and -DQB1 alleles were determined by DNA typing 50 patients and 108 controls. HLA distributions were analysed in the following autoantibody subgroups: anti-SS-A with -SS-B, anti-SS-A without -SS-B, anti-snRNP without -Sm, anti-SS-A without -snRNP and anti-snRNP without -SS-A. The autoantibodies were detected by EIA (enzyme immunuassay). Patients with anti-SS-A and -SS-B had significantly increased frequencies of DRB1*03, DRB3*0101, DQA1*0501, DQB1*0201 (in linkage disequilibrium) versus controls and versus patients without anti-SS-A and -SS-B. No differences in HLA distribution were found when the group with anti-SS-A alone was compared to the group with anti-SS-A and concomitant -SS-B. Comparing the groups with and without anti-SS-A and -SS-B, the highest RR were found for the alleles DRB1*03, DRB3*0101, DQB1*0501, DQB1*0201 (in linkage disequilibrium) with RR: 16.8, 5.0, 19.6, 10.3, respectively, P < 0.05). RR for DQw2/DQw6 heterozygotes was 3.5 (Ns.), and RR for cases having DQ alpha molecules with glutamine in position 34 and DQ beta molecules with leucine in position 26 on both chains was 6.3 (P < 0.05). No HLA associations were observed in the group with anti-snRNP without concomitant -Sm or without concomitant -SS-A. These results show that production of anti-SS-A and -SS-B is associated to the HLA alleles DRB1*03, DRB3*0101, DQA1*0501, DQB1*0201, and that this haplotype shows stronger correlation to these responses than DQw2/DQw6 heterozygosity or HLA molecules having glutamine in position 34 (DQ alpha) and leucine in position 26 (DQ beta). The failure to observe any correlation with DRBI*15,16 (DR2) in the group with anti-SS-A alone may demonstrate ethnic differences concerning this response. The failure to identify any HLA associations for the anti-snRNP response may reflect the heterogeneity of the molecules that constitute this antigen. PMID- 7481564 TI - The importance of C4A null genes in Norwegian patients with systemic lupus erythematosus. AB - C4A null genes were determined by RFLP (Taq I) and SSO-probing on PCR-amplified C4-DNA in 51 Scandinavian patients with systemic lupus erythematosus (SLE) and 124 controls. Associations of the alleles DRB1*0301, DQA1*0501, DQB1*0201 had previously been found in this SLE group, as well as increased frequency of HLA DRB1 and -DQ homozygosity. The frequency of the allele C4A*Q0 was increased among the patients (RR = 2.3, P = 0.0172). The SSO-probing revealed additional cases of C4A*Q0 homozygotes among the controls, leading to diverging RR values for C4A*Q0 homozygotes depending on the technique used. The RFLP method gave an RR of 9.7 (P = 0.0028), while the SSO-probing resulted in an RR of 4.8 (P = 0.0153), demonstrating that unprecise characterization of C4A*Q0 in a relatively small material has great effect on the calculated RR. Multiple 2 x 2 tests were performed in an attempt to detect the strongest association of the alleles DRB1*0301, DQA1*0501 and C4A*Q0 (in linkage disequilibrium). These comparisons showed a trend towards stronger association for DAQ1*0501 and DRB1*0301 than for C4A*Q0, and no interaction between the HLA alleles and the allele C4A*Q0. This may suggest that HLA class II molecules themselves and/or an unknown susceptibility gene located near the DQA1 and DRB1 loci are involved in the pathogenesis of SLE. PMID- 7481565 TI - Correlation of IgG Fc receptors on granulocytes with serum immune complex level in systemic lupus erythematosus. AB - The expression of Fc gamma RI, Fc gamma RII and Fc gamma RIII on granulocytes in the blood of patients with systemic lupus erythematosus was investigated. The relationship between the receptor expression and serum immune complex (IC) concentration was analysed. The decrease in mean fluorescence intensity of both Fc gamma RII and Fc gamma RIII of patients' granulocytes stained by specific monoclonal antibodies (using MoAb IV.3 and 3G8) was significant. The detected decrease of Fc gamma RII was inversely correlated with the high circulating IC level in patients' sera. PMID- 7481566 TI - Late-onset schizophrenia and late paraphrenia. AB - The term "late-onset schizophrenia" was first coined by Manfred Bleuler (1943) to describe a form of schizophrenia with an onset between the ages of 40 and 60. This concept has been adopted by German psychiatry. Until recently, British and American psychiatrists had little interest in this patient group. However, they often used the term "late-onset schizophrenia" interchangeably with late paraphrenia or as a generic term for both these diseases, even though the concept of late paraphrenia is quite different from that of late-onset schizophrenia. Late paraphrenia is a British concept that includes all delusional disorders starting after age 60. This confusion of terms and concepts is even more important now, because recent neuroimaging and neuropsychological studies suggest that an organic substrate probably exists in most cases of late paraphrenia, while only minor organic abnormalities can be found in late-onset schizophrenia. We believe it is of utmost importance to establish a clear boundary between late onset schizophrenia and other delusional disorders in middle and old age, because the confusion in terminology and concepts is a serious impediment to comparative international research. PMID- 7481567 TI - Prenatal development of monozygotic twins and concordance for schizophrenia. AB - While twin concordances for schizophrenia have been used to estimate heritability and to develop genetic models, concordances in subtypes of monozygotic (MZ) twins can also be used to investigate the influence of prenatal development in the etiology of mental illness. We used within-pair variability and mirroring of fingerprints to estimate retrospectively the placentation status of concordant and discordant MZ twins. The results indicate that concordant MZ pairs were more likely to have been monochorionic (MC) and to have shared a single placenta, whereas discordant MZ pairs appear more likely to have been dichorionic (DC) with separate placentas. Pairwise concordances for MZ twins without MC markers averaged 10.7 percent. In contrast, concordances for MZ twins with one or more MC markers averaged 60 percent. This suggests that simple MZ concordance rates may overestimate schizophrenia heritability and that prenatal development may also be important in the etiology of schizophrenia. Because MC (but not DC) twins usually share fetal blood circulation and hence are likely to share infections, these results are consistent with the hypothesis that fetal infections may be a significant etiological factor in schizophrenia. PMID- 7481568 TI - Expectations and motives for substance use in schizophrenia. AB - This study examined the internal reliability of standardized measures of substance use expectancies and motives in a schizophrenia population (n = 70) and the relationship of these expectancies and motives to alcohol and drug use disorders. Internal reliabilities were uniformly high for the subscales of the expectancy and motive measures. Analyses of the relationship between substance use disorders and expectancies revealed strong substance-specific expectations. Alcohol expectancies were related to alcohol disorders but not to drug disorders; cocaine expectancies were related to drug but not to alcohol disorders; and marijuana expectancies were more strongly related to drug than to alcohol use disorders. In contrast, motives were related to substance use disorders, and self reported substance use problems were related to expectancies and motives in a non specific manner. These results suggest that expectancy and motive questionnaires developed for the primary substance abuse population may be valid for psychiatric populations. Research on motives and expectancies may help to clarify the functions of substance abuse in persons with schizophrenia. PMID- 7481569 TI - Personal therapy: a disorder-relevant psychotherapy for schizophrenia. AB - While the long-term care of ambulatory schizophrenia patients requires highly effective interpersonal treatment skills among clinicians, there is little evidence to support an empirically validated individual psychotherapy of schizophrenia. Personal therapy (PT) attempts to address the apparent limitations of traditional psychotherapy by modifying the "model of the person" to accommodate an underlying pathophysiology, minimizing potential iatrogenic effects of maintenance antipsychotic medication, controlling sources of environmental provocation, and extending therapy to a time when crisis management has lessened and stabilization is better ensured. By means of graduated, internal coping strategies, PT attempts to provide a growing awareness of personal vulnerability, including the "internal cues" of affect dysregulation. The goals are to increase foresight through the accurate appraisal of emotional states, their appropriate expression, and assessment of the reciprocal response of others. The strategies are supplemented by phase-specific psychoeducation and behavior therapy techniques. Practical issues in the application of this new intervention are discussed. Preliminary observations from two samples of patients, one living with and the other living independent of family, suggest differential improvement over time among PT recipients. PMID- 7481570 TI - Interpersonal problem solving and information processing in schizophrenia. AB - Cognitive models of interpersonal problem solving have been proposed for, but infrequently tested on, samples of schizophrenia subjects. This study undertook to examine the relationships between the receiving, processing, and sending skills that comprise one model of interpersonal problem solving with information processing and social cue perception. Twenty-six patients with a DSM-III-R diagnosis of schizophrenia or schizoaffective disorder completed measures of interpersonal problem solving, social cue perception, visual vigilance, verbal memory, conceptual flexibility, and psychiatric symptoms. Significant and robust relationships were found between sensitivity to social cues and receiving, processing, and sending skills. Only recognition and recall memory, of the various other information-processing measures, were found to be related to any of the three problem-solving skills. Associations between problem solving and cognitive deficits did not seem to be attributable to psychiatric symptoms. Implications of these findings for understanding and remediating the problem solving deficits of schizophrenia patients are discussed. PMID- 7481571 TI - Psychosocial functioning and subjective experience in schizophrenia: a reanalysis. AB - Data collected by Brekke et al. (1993) on the symptomatology, psychosocial functioning, and subjective experience of schizophrenia outpatients were reanalyzed using LISREL to elucidate a causal model that would depict the functional relationships between the variables. The model that best fit the data parallels another model tested previously on cardiac patients and shows that subjective experience is much more influenced by symptomatology than by social functioning. This confirms Brekke et al.'s main finding. The implications of these results for intervention and for future research are considered. PMID- 7481572 TI - Expressed emotion and language disturbances in parents of stable schizophrenia patients. AB - Nineteen healthy parents of long-term schizophrenia outpatients were assessed for levels of expressed emotion (EE) and for characteristics of communication style which are putative markers of vulnerability to schizophrenia. We administered measures of communication deviance, linguistic reference performance, global disorganization, and positive formal thought disorder. Parents high in EE showed significantly poorer linguistic reference performance and greater disorganization in their speech than parents low in EE. These findings support the idea that high EE in some individuals may be associated with cognitive characteristics indicative of a vulnerability to schizophrenia, and this may account in part for the well-established association between EE level in parents and prognosis in patients. PMID- 7481574 TI - The epidemiology of drug-induced akathisia: Part I. Acute akathisia. AB - This article reviews the epidemiological data on drug-induced acute akathisia, examining studies in which akathisia was the primary focus as well as those in which it was one of a number of drug side effects studied. The studies are diverse in methodology and suffer from many limitations. Incidence rates for acute akathisia with conventional neuroleptics vary from 8 to 76 percent, with 20 to 30 percent being a conservative estimate; preliminary evidence suggests that the newer atypical antipsychotic drugs are less likely to produce acute akathisia. A number of nonneuroleptic drugs--in particular the serotonin-specific reuptake inhibitors--have been implicated in the development of akathisia, but the epidemiological data are limited. Risk factors for neuroleptic-induced akathisia are not completely understood. Drug dose, rate of increment of dose, and drug potency seem to be important, but the role of sociodemographic factors and other treatment-related variables is modest. Drug-induced parkinsonism is significantly correlated with akathisia. Evidence for iron deficiency as a risk factor is conflicting, and its contribution is likely to be minor. PMID- 7481573 TI - Cost of relapse in schizophrenia. AB - To estimate the national annual cost of rehospitalization for multiple-episode schizophrenia outpatients, and to determine the relative cost burden from loss of medication efficacy and from medication noncompliance, the yearly number of neuroleptic-responsive multiple-episode schizophrenia inpatients in the United States who are discharged back to outpatient treatment was estimated. The cohort at risk for future relapse and rehospitalization was determined. The research literature on the expected rates of relapse for schizophrenia patients on maintenance antipsychotic medication was reviewed; in particular, monthly relapse rates under the optimal medication conditions of compliant patients taking optimal doses of a depot neuroleptic (optimal neuroleptic dose) and under the less optimal conditions of patients stopping medication (medication noncompliant) was estimated. Using established noncompliance rates from the literature, it became possible to estimate a "real world" rehospitalization rate for this cohort, as well as the relative burden accruing from loss of medication efficacy and from medication noncompliance. Finally, cost estimates for index hospitalizations and rehospitalizations were derived from data on national expenditures for inpatient mental health care. The monthly relapse rates are estimated to be 3.5 percent per month for patients on maintenance neuroleptics and 11.0 percent per month for patients who have discontinued their medication. Postdischarge noncompliance rates in community settings are estimated to be 7.6 percent per month. These estimates were entered into a survival analysis model to determine the real world relapse rate of this cohort. An estimated 257,446 multiple-episode (> or = two hospitalizations) schizophrenia patients were discharged from short-stay (< or = 90 days) inpatient units in the United States during 1986. The estimated aggregate baseline inpatient cost for the index hospitalizations of this cohort was $2.3 billion (1993 dollars). Within 2 years after discharge, the aggregate cost of readmission approached $2 billion. Loss of neuroleptic efficacy accounted for roughly 60 percent of the rehospitalization costs and neuroleptic noncompliance for roughly 40 percent. The economic burden due to loss of efficacy is relatively higher during the first postdischarge year, whereas the burden from noncompliance is higher in the second year. Because loss of medication efficacy and medication noncompliance act synergistically on relapse, substantial inpatient cost savings can be realized by linking better pharmacologic treatments of schizophrenia with more effective strategies to manage medication noncompliance. PMID- 7481575 TI - The epidemiology of drug-induced akathisia: Part II. Chronic, tardive, and withdrawal akathisias. AB - This article examines the epidemiological data on chronic akathisia, tardive akathisia, and withdrawal akathisia. The limitations of the data are discussed- in particular, the lack of consistent definitions of the syndromes. The studies suggest that a significant proportion of patients chronically treated with neuroleptics suffer from akathisia. The prevalence may be as high as 40 percent, although a conservative estimate would be closer to 30 percent. Risk factors for the development of chronic akathisia and tardive akathisia are poorly understood, but old age, female sex, iron deficiency, negative symptoms, cognitive dysfunction, and affective disorder diagnosis need to be studied further for their potential role. While there is convincing evidence that akathisia may develop after neuroleptic cessation or reduction in dose, the prevalence and risk factors for withdrawal akathisia are not known. Reports of akathisia in children and the elderly have been few, and more systematic research is necessary. Akathisia appears to be common in individuals with mental retardation treated chronically with neuroleptics. PMID- 7481576 TI - Weight gain associated with neuroleptic medication: a review. AB - In this article we review the empirical literature on weight gain associated with neuroleptic drug use. Weight gain, which appears to be associated with an increase in appetite, is variable but likely to be larger initially and then plateau. Clozapine and low-potency phenothiazines are associated with the largest gains and molindone with weight loss, but the mechanism is not known. Amantadine and fenfluramine may reverse weight gain to some degree. Dietary fat seems to play an important role in obesity, and research is needed to increase the data base and elucidate possible mechanisms. Studies are also needed to evaluate preventive strategies and to determine which drugs are least likely to produce weight gain as well as which drugs could be added to a neuroleptic regimen to control weight. PMID- 7481577 TI - Psychopathological dimensions in schizophrenia. AB - To test underlying latent syndromes within schizophrenic syndromes assessed by the Positive and Negative Syndrome Scale (PANSS), a sample of 100 DSM-III-R schizophrenia patients was obtained through a semistructured interview for schizophrenia. Confirmatory factor analysis was used to evaluate the underlying symptomatological dimensions. The positive and negative construct was not confirmed by this study. The three- and four-dimension models obtained higher goodness of fit than the one- or two-dimension models. Models composed of three or four dimensions shared positive and negative syndromes, in addition to "disorder of relating" and/or "disorganization" dimensions. The disorder of relating dimension of PANSS was composed of emotional withdrawal and passive/apathetic social withdrawal. The disorganization dimension comprised two symptoms of PANSS (conceptual disorganization and poor attention) and an item not included in the PANSS: inappropriate affect. It is suggested that a schedule for the assessment of this item be added to the PANSS. PMID- 7481578 TI - Time trends in schizophrenia: changes in obstetric risk factors with industrialization. AB - The frequency of schizophrenia may have increased during the early stages of industrialization and may now be declining. Early in the process of development, the illness appeared to be more common among the upper classes and later, more common in the lower classes. As with certain other diseases, the occurrence of schizophrenia may be influenced by the transition from poverty to affluence. Explanations for such a pattern of occurrence include the combined effect of (1) variation in the rate of birth complications owing to cephalopelvic disproportion secondary to changes in nutrition, and (2) increased infant survival following improvements in obstetric and neonatal care. These effects help elucidate the pattern of occurrence of schizophrenia in immigrant groups. PMID- 7481579 TI - First person account: schizophrenia--adrift in an anchorless reality. AB - The article that follows is part of the Schizophrenia Bulletin's ongoing First Person Accounts series. We hope that mental health professionals--the Bulletin's primary audience--will take this opportunity to learn about the issues and difficulties confronted by consumers of mental health care. In addition, we hope that these accounts will give patients and families a better sense of not being alone in confronting the problems that can be anticipated by persons with serious emotional difficulties. We welcome other contributions from patients, ex patients, or family members. Our major editorial requirement is that such contributions be clearly written and organized, and that a novel or unique aspect of schizophrenia be described, with special emphasis on points that will be important for professionals. Clinicians who see articulate patients, with experiences they believe should be shared, might encourage these patients to submit their articles to First Person Accounts, Division of Clinical and Treatment Research, NIMH, 5600 Fishers Lane, Rm. 18C-06, Rockville, MD 20857. PMID- 7481580 TI - Systemic sclerosis 'state of the art' 1995. PMID- 7481581 TI - Cytokine autoantibodies in rheumatoid arthritis. AB - Sera from 42 patients with rheumatoid arthritis (RA) and 40 healthy controls (HC) were examined for cytokine autoantibodies (CK-aAb) by accurate and sensitive radioimmunoassays. The prevalences of detectable CK-aAb in RA (HC) were: aAb-IL-1 alpha = 36% (38%); aAb-IL-6 = 29% (13%), p = 0.06; aAb-IL-8 = 0% (0%); aAb-IFN alpha = 12% (3%), p = 0.11. The levels of the individual CK-aAb did not correlate, and there were no correlations between CK-aAb levels and clinical or laboratory variables. CK-aAb levels remained constant in 8 RA patients tested over a period of 6 months. With regard to alterations in aAb-IL-1 alpha levels, 4/11 HC were consistently positive over 18-36 months; 2/11 converted and became highly positive. The levels of aAb-IFN alpha and aAb-IL-6, but not aAb-IL-1 alpha, tended to be increased in RA patients; aAb-IL-8 were undetectable in both RA and HC. PMID- 7481582 TI - Serum values of pepsinogens and antibodies to Helicobacter pylori in relation to a history of peptic ulcer disease and NSAID use. AB - The present study was undertaken to investigate whether patients with a history of peptic ulcer disease (PUD) have serological features indicative of the presence of more severe gastritis, compared to patients without a history of PUD. In addition we investigated whether current chronic use of non steroidal anti inflammatory drugs (NSAIDs) was associated with serological features indicative of a chemical (type C) gastritis. In total 850 consecutive outclinic patients were interviewed by a standard questionnaire with emphasis on demographic data, diagnosis, chronic use of NSAID, and history of PUD. Serum pepsinogen A (PgA) and pepsinogen C (PgC), the PgA:PgC ratio, and IgA and IgG antibodies to H. pylori were measured in all patients. After controlling for age, sex, H. pylori seropositivity, and presence of RA, a correlation was found between a decreased pepsinogen A:C ratio and the use of NSAID. Patients with a history of PUD have serological features indicative of a persistently more severe gastritis. Secondly we found serological evidence for the existence of a chemical (type C) gastritis in NSAID users. PMID- 7481583 TI - Antibodies against oxidised low-density lipoprotein in juvenile chronic arthritis. AB - Enhanced lipid peroxidation, reported to take place in rheumatoid joints and suggested to play a significant role in joint inflammation, led us to study the occurrence of antibodies against oxidised low-density lipoprotein (Ox-LDL Ab) in patients with juvenile chronic arthritis. Enzyme-linked immunosorbent assay was used to detect Ox-LDL Ab and antiphospholipid antibodies (aPL Ab) in sera from 84 patients and 91 controls. Elevated levels of Ox-LDL Ab were found in 14 patients (17%) as opposed to 4 controls (4%; p < 0.01). Similarly, 14 patients had an elevated aPL Ab level and a fairly good correlation between Ox-LDL Ab and aPL Ab (r = 0.52) existed in the patients. The increased frequency of elevated levels of Ox-LDL Ab may reflect lipid peroxidation occurring in rheumatoid joints but crossreactivity with aPL Ab for the induction of Ox-LDL Ab cannot be excluded. PMID- 7481584 TI - Scoring of synovial membrane hypertrophy and bone erosions by MR imaging in clinically active and inactive rheumatoid arthritis of the wrist. AB - MRI-scores of synovial membrane hypertrophy and bone erosions of the RA-wrist are introduced. Gadolinium-DTPA enhanced magnetic resonance imaging (MRI) and conventional radiography (CR) of the wrist were performed in 16 patients with rheumatoid arthritis (RA) and 3 healthy controls. A MRI-score of synovial membrane hypertrophy was obtained by summation of gradings of synovial hypertrophy in 6 regions of the wrist. The score was significantly higher in wrists with than in wrists without clinical signs of active arthritis. The score was 0 in all healthy controls. Each bone of the wrist was assessed by MRI and CR with respect to bone erosions. Bone erosions were detected by MRI in 14 wrists in contrast to only 6 wrists by CR. In all patients the erosions were more numerous on MRI. The introduced methods may be useful quantitative measures of synovitis and early joint destruction in RA. PMID- 7481586 TI - HLA-B27 in healthy adults in UAE. An extremely low prevalence in Emirian Arabs. AB - We reviewed the prevalence of HLA-B27 in 760 healthy individuals in the UAE, where the community is multi-ethnic. All were potential live donors for renal and bone marrow transplantation. The overall prevalence rate was 6.4%; 5.7% in Arabs only and 7.4% in the Asian group. Significant intra-racial variation were not observed between the major ethnic groups (P 0.235 and 0.1). Emirian Arabs exhibited a very low prevalence rate of 0.5% which was highly significant in comparison to other Arab subgroups, thus allowing a degree of intra-racial variations of the marker. The pair-wise comparison in all other Arab subgroups did not yield any significant differences. The Yemeni Arabs in this study had the highest rate of 17% and unexpectedly, Arabs with an African admixture (Somalis and Sudanese) also showed relatively high figures. PMID- 7481585 TI - Plasma viscosity in the monitoring of therapy in rheumatoid arthritis patients. AB - Plasma viscosity (PV) was compared with erythrocyte sedimentation rate (ESR), C reactive protein (CRP) and clinical measures in the monitoring of parenteral gold therapy in a longitudinal study of 40 consecutive rheumatoid arthritis (RA) patients. Significant correlations with the combined clinical variables were only found for differences in PV and not for the other laboratory tests. PV but not ESR or CRP discriminated significantly between active and inactive disease. The sensitivity and specificity for PV above the normal range as a sign of disease activity was 89% and 73%, respectively, which was superior compared with ESR and CRP. The advantages of PV over CRP and ESR justify wider use of this test and warrant further studies of PV in the monitoring of other therapies in RA patients. PMID- 7481587 TI - Arthrodesis in rheumatoid arthritis using absorbable screws and rods. AB - In this series absorbable fixation devices (screws and rods) made of self reinforced poly-L-lactide (SR-PLLA) were used to stabilize arthrodesis in rheumatoid arthritis. These fixation devices have been used mainly in the fracture treatment or in the fixation of osteochondral lesions or osteotomies in hallux valgus operations. In arthrodesis of post-traumatic arthritis these devices have been used successfully. Fifty-three arthrodeses (18 in the wrist, 18 in the hand, 6 talocrural joint and 11 subtalar-calcaneocuboid-talonavicular joint) were performed for 47 patients using absorbable screws and rods between 1989 and 1994. Overall results in every group were good. There were 3 superficial infections and 2 nonunions (both talocrural arthrodesis). According to this study it seems that absorbable screws and rods are interesting alternative fixation devices in arthrodesis in rheumatoid arthritis. PMID- 7481588 TI - Rheumatic pains of previously undiagnosed diabetic subjects. AB - To identify the early diabetic musculoskeletal symptoms of previously undiagnosed diabetic subjects, a case-control study was carried out. The cases and controls were recruited from a population aged 55 years. Questions concerning the symptoms were asked before the 2-h oral glucose tolerance tests (OGTT). The results show that pain in the right hand was the most prominent symptom among the diabetic women. Pains in the left hand and the shoulders in the diabetic women and pains in the right knee and the right hip joint in the diabetic men tended to be more prevalent than the corresponding symptoms in the controls. The highest prevalence of most musculoskeletal pains occurred in the highest tertile of 2-h OGTT values among women. The conclusion is that the hand pain is closely associated with the development of diabetes and may give clues to an early diagnosis of diabetes in a middle-aged population. PMID- 7481589 TI - The reliability and validity of pain threshold measurements in osteoarthritis of the knee. AB - The purpose of this study was to examine the reliability and validity of measuring pain threshold (PT) of persons with osteoarthritis (OA) of the knee. The PTs of women with and without OA of the knee were measured on 3 occasions at 6 sites at the knee with a pressure dolorimeter. Subjects with OA also recorded their pain on a visual analogue scale (VAS) and on the McGill Pain Questionnaire (MPQ). The reliability coefficients for repeated measures of PT varied from 0.61 to 0.91 in the OA group and from 0.71 to 0.90 in the control group. PT was lower in the OA group at all sites. Only the control group demonstrated significant differences between sites. The correlations between PT and measures of pain intensity were poor. It is concluded that the measurement of PT at the knee distinguishes OA from healthy controls, and that it has moderate reliability. However, it cannot substitute for a measure of pain intensity. PMID- 7481590 TI - Are tender point scores assessed by manual palpation in fibromyalgia reliable? An investigation into the variance of tender point scores. AB - To determine the reliability of tender point assessment by manual palpation in patients with fibromyalgia, and to investigate the sources of variance, tender point scores by manual palpation were obtained in 30 fibromyalgia patients by 2 investigators; the assessments were repeated after 1 week. Test-retest stability and inter-observer agreement of tender point scores was moderate to high. The most important sources of variance were inter-individual patient differences in (1) mean tenderness (pain thresholds) and (2) tender point profiles (consistent intra-individual differences between the 14 tender point scores), and (3) residual variance, not attributable to any systematic effect (error). It is concluded that the reliability of tender point assessment by manual palpation is acceptable. It seems to be not less than reliability of assessment by pressure algometer. Tender point scores in fibromyalgia reflect individual differences in mean pain thresholds as well as individual tender point profiles. PMID- 7481591 TI - Fungal arthritis. A review. AB - Fungal arthritis (FA) due to a candida infection was reviewed in English literature and described in 45 cases. The increasing use of potent antibiotics, immunosuppressives, and especially the use of artificial joints, predisposed to the infection. Weightbearing joints, particularly the knees, were most frequently affected. Symptoms were described as a warm, tender and swollen joint, and the duration of symptoms prediagnosis was described as being up to 4 years. Amphotericin B was the treatment drug of choice, and in cases of no response, supplemented with either flucytosine or ketoconazole. Local FA healed in all cases. FA in an artificial joint resulted in all cases in removal of the prostheses. Mortality in systemic fungal infections including FA was 50%. PMID- 7481593 TI - Adhesive capsulitis. Is it possible in childhood? AB - Adhesive capsulitis affecting shoulders has been extensively studied by orthopedic surgeons, but few reports have been published on this condition when it affects other joints (hips, ankles). To our knowledge no pediatric cases have been reported. As its treatment requires prolonged physiotherapy and sometimes manipulation, a correct diagnosis is essential. We would like to emphasize the importance of having this condition in mind when one is regarding a differential diagnosis, also in children. PMID- 7481594 TI - Small nerve fibre dysfunction in a patient with Sjogren's syndrome. Neurophysiological and morphological confirmation. AB - An morphological confirmation of neurophysiologically assessed small nerve fibre dysfunction in a patient with Sjogren's syndrome was sought. Conventional motor and sensory nerve conduction studies were normal, while examination of the thermal specific and thermal pain sensitivity and an autonomic nervous system evaluation showed striking abnormalities. Sural nerve electron microscopy showed almost normally large nerve fibres and abnormalities in unmyelinated nerve fibres or Schwann cells, and verified morphologically the neurophysiologically supposed small nerve fibre involvement. PMID- 7481592 TI - Thymoma and pure red cell aplasia in a patient with systemic lupus erythematosus. AB - We present the case of a female patient with a diagnosis of systemic lupus erythematosus (SLE) at the age of 54 years. At the age of 63 years, she suffered from malignant thymoma and 3 years after removal of the thymoma a diagnosis of pure red cell aplasia (PRCA) was established. This is, to our knowledge, the first report of the occurrence of SLE, thymoma and PRCA in the same patient. The case is discussed with regard to the already known associations between these diseases. PMID- 7481595 TI - Pulmonary aspergilloma complicating Wegener's granulomatosis. PMID- 7481596 TI - Sex role orientation and its relationship to the development of identity and moral thought. AB - This paper examined the links among sex role orientation, ego identity development and moral reasoning. 76 female and 58 male students were assessed on Bem's sex role inventory, ego identity, care-based and justice-based moral thought. For women, identity was negatively related to femininity and positively related to androgyny. Also, high levels of care-based moral reasoning, i.e., a balanced concern for the welfare of self and others, was related to androgyny for women. There were no significant findings for men. No significant relationships were found between sex role orientation and justice-based moral reasoning. Thus, the results suggested that for women in particular, relinquishing the stereotyped sex role definition and becoming more androgynous is important for their ego identity as well as care-based moral development. PMID- 7481597 TI - Cognitive sequelae of repeated head injury in a population of intravenous drug users. AB - The relationship between closed head injury and performance on neuropsychological (NP) tests was investigated in a group of intravenous drug users (IVDUs). Subjects with repeated head traumas involving loss of consciousness (LOC) performed worse than both a control group without LOC and reference group with only a single episode of LOC. There were no significant differences between the last two groups. Performance on tests of memory, attention, and motor performance was significantly worse in the group with repeated head injury. The average time since the last episode of LOC was more than 11 years. We conclude from these findings that a single episode of LOC does not result in significant cognitive impairment in this population. Two or more episodes, however, are more likely to produce chronic cognitive impairment. PMID- 7481598 TI - Personality disorders and psychosocial changes after intensive psychotherapy: a prospective follow-up study of an outpatient psychotherapy project, 5 years after end of treatment. AB - This report presents the results of a 7-year prospective outcome study designed to examine the psychosocial changes during and after therapy among 25 outpatients suffering from personality disorders and psychoses. The therapeutic approach was based mainly on object relations theory and psychodynamic self-psychology, and focused in particular on affect consciousness, parental images, self-image and interpersonal relations. Twenty patients completed the form which measured psychosocial changes during therapy, and 21 persons participated at follow-up. The data show statistically significant changes in the capacity to tolerate intimate relationships and actually establish such relationships, improved quality of contact in relationships with friends, a general raising of socioeconomic status and reduced use of ordinary health and social services. Both the global psychosocial outcome at follow-up as measured by HSRS and the general level of symptoms measured by SCL-90 suggest that 76% of the sample had reached a level of psychosocial functioning and adaptation that can be defined as "no caseness". PMID- 7481599 TI - RASP: a system for the analysis of memory tasks. AB - The purpose of this paper is to introduce a system for the analysis of memory tasks. The system involves four basic task dimensions, derived from an ecological analysis of memory functioning: Reproductivity (R), Acquisition (A), Sufficiency (S), and Productivity (P). Three studies are reported in which professional memory psychologists were asked to rate sets of standard laboratory and more ecologically relevant memory tasks in terms of the four RASP dimensions. In general, results implied independence across the two sets of tasks between the four dimensions. Interrater reliabilities were found to be modest to high. However, it was suggested that RASP best be used as a tool for understanding the meaningful relations that exist among dimensions within general sets or pools of tasks, for different classes of tasks, as well as for prototypical tasks. We argue that, especially when the relationship between RASP profiles for two particular tasks is crossvalidated by behavioral data, and conflict with assumptions about what memory systems or processes are involved, then there is a potential for conceptual refinement. This refinement could be attained either through an increased theoretical generalization, or through an increased theoretical precision. Based on obtained RASP information at the level of classes of tasks (e.g., episodic vs. semantic) as well as at a prototypical task level (e.g., word recall vs. prose recall), several intriguing memory phenomena are discussed. Finally, we suggest that RASP may serve as a tool for reducing the number of task-specific assumptions of what systems and processes mediate memory performance. PMID- 7481601 TI - Behavioral determinants of mothers' safety measures to prevent injuries of pre school children. AB - Home-related injuries are a major threat to pre-school children in the western world. In this study the behavioral determinants of 18 parental safety measures were assessed. To select behavioral determinants, the Attitude-Social influence Self-efficacy/barriers model was used with the inclusion of variables from the Health Belief Model and the Protection Motivation Theory. A written questionnaire was completed by 1129 Dutch mothers of pre-school children. Most safety measures were explained (rather) well by the same set of determinants. Main determinants for adopting or not adopting a safety measure were the mother's belief in the necessity of the safety measure according to the child's age, her belief about her partner's opinion on the necessity and her belief about the success of taking the measure. Subsequent important determinants were the mother's belief about the inconvenience and instrumentality of the safety measure; the perceived susceptibility had a minor or moderate influence on most measures. This study did not indicate that more highly educated mothers were more knowledgeable about safety-related subjects. The theoretical implications of this study are discussed. PMID- 7481600 TI - Work motivation among high school students before and during the economic recession in the Swedish labour market. AB - In 1990, 2349 high school pupils from 21 high schools in the Stockholm, Uppsala and Sodertalje regions answered a questionnaire concerning how important different characteristics of work were in their future job. The questionnaire also included an open question, "How would you describe an ideal job?" In 1993 the same questionnaire was answered by 1171 high school students. The two measurement occasions were separated by economic changes leading, from a condition of economic security and full employment, to a condition characterised by economic insecurity and high unemployment especially among young people. Results showed that the economic insecurity of 1993 lowered the students' ratings of the importance of all characteristics of work with the exception of the security of work. A strong gender factor influenced the ratings. Thus, the female students rated the importance of the characteristics of work higher than the male students did. The only exception concerned "good pay", which was rated higher by the male. In 1993 these effects were less pronounced, although still evident. The motivational structure seemed to be unaffected by the economic insecurity. The same four motivational factors, i.e. (1) the expressive work goals, (2) the working conditions, (3) the benefits of work and (4) the social aspects of work, appeared in the analyses of both measurement occasions. PMID- 7481602 TI - Preferential looking: a sudden shift in visual fixation pattern at 32 months of age. AB - Observations of eye movements of young children in a modified preferential looking task suggest a change in the visual looking pattern taking place during a very brief time between 32 and 33 months of age. In the younger children, a grating stimulus elicited eye movements towards the target followed by a few seconds of focused attention; in the older children a visual avoidance behavior was observed where subjects looked consistently in the opposite direction of the target. An analysis of video recordings suggests that this avoidance pattern was a composite of a very brief initial target-directed eye movement which was quickly arrested, and followed by eye movements in the opposite direction, initiating a search of the visual scene. PMID- 7481603 TI - Research, prevention and impact. PMID- 7481605 TI - Pooled reanalysis of cancer mortality among five cohorts of workers in wood related industries. AB - OBJECTIVES: To provide more information regarding the risk of cancer associated with wood dust, a pooled reanalysis of data from five cohort studies was performed. METHODS: The combined cohort consisted of 28,704 persons from five studies: British furniture workers, members of the union representing furniture workers in the United States, two cohorts of plywood workers, and one of wood model makers, among whom 7665 deaths occurred. Pooled analyses were carried out for all of the cohorts combined, the two furniture worker cohorts combined, and the two plywood workers cohorts combined. RESULTS: Significant excesses of nasal [observed 11, standardized mortality ratio (SMR) 3.1, 95% confidence interval (95% CI) 1.6-5.6] and nasopharyngeal (observed 9, SMR 2.4, 95% CI 1.1-4.5) cancer were observed. That for nasal cancer appeared to be associated with exposure to wood dust but was based solely on cases from the British furniture worker cohort, while that of nasopharyngeal cancer was observed for furniture and plywood workers and was associated with both high and low probability of wood dust exposure. Some support for an excess risk of multiple myeloma was also observed but was less clearly associated with wood dust exposure. No excesses of lung, larynx, stomach, or colon cancer were found to be associated with any surrogate indicators of wood dust exposure. CONCLUSIONS: Workers exposed to wood dust may have an excess risk of nasopharyngeal cancer and multiple myeloma in addition to sinonasal cancer. The limitations of this study would tend to obscure relationships, rather than create false positive findings. PMID- 7481604 TI - Possible mechanisms for the respiratory tract effects of noncarcinogenic indoor climate pollutants and bases for their risk assessment. AB - This review outlines the effects of pollutants on the lungs. Mechanisms and effects relevant to the assessment of indoor-air risk are especially dealt with. Important mediators have also been considered. Concentration-effect relationships exist for toxic reactions, sensitization reactions, and neurogenic effects. If Harber's Law is used for extrapolations from higher concentrations to the lower indoor-air levels, the indoor-air risk estimate may exceed the real risk. Additivity seems to apply to toxic and neurogenic effects of low doses. Only already sensitized subjects and possible subjects with a profound alpha 1 antitrypsin deficiency appear to be extremely sensitive, and a safety factor of 10 seems adequate for the protection of other groups. Thus combining occupational exposure limits (OEL), Harber's Law, and the safety factor suggests that no direct lung effects should be expected from a substance if the exposure level does not exceed 1/40.OEL. PMID- 7481606 TI - Pregnancy-related sickness absence among employed women in a Swedish county. AB - OBJECTIVES: The objective of this study was to analyze the variation of pregnancy related sickness absence among employed women according to age, occupation, and degree of male-female domination within occupations. METHODS: Data from a prospective study of all new sick-leave spells exceeding 7 d in 1985 and 1986 in the county of Ostergotland, Sweden, were related to the population at risk, through the Swedish Medical Birth Register. The subjects included in the analysis were all 7000 employed women that gave birth in 1985 and 1986, of which some 3000 were sick-listed at least once with pregnancy-related diagnoses. RESULTS: There was little difference in the pregnancy-related sickness absence between the age groups. The age-standardized rate for sick leaves involving pregnancy-related diagnoses differed substantially between occupations. Women in the metal industry had the highest rates; those employed in administration, banking, and insurance had the lowest. White-collar occupations generally had lower rates and blue collar occupations higher rates, with some exceptions (eg, in saw mills, farming, and the chemical industry). Gender-integrated occupations had the lowest sick leave rate, while extremely male-dominated jobs had the highest. The latter association remained after adjustment for occupational area. CONCLUSIONS: There were considerable differences between occupational groups in the rates of sick leaves involving pregnancy-related diagnoses. Some differences were related to physical load of the jobs being done, but not all. It seems important to consider also male-female domination within a job with respect to such sick leaves. PMID- 7481607 TI - Influence of occupational physical activity on pregnancy duration and birthweight. AB - OBJECTIVES: The influence of occupational physical activity on pregnancy duration and birthweight was examined. METHODS: In this prospective study information on levels of occupational physical activity was collected during a personal interview before pregnancy, and possible changes were registered during follow up, which lasted until after birth. Data on pregnancy duration and birthweight were obtained from midwives, physicians, and obstetricians. The occupational energy expenditure was operationalized in intensity and fatigue scores, which were studied as such and in combination with workhours and work speed. The occupational biomechanical load was operationalized in a peak and a chronic pressure score. RESULTS: The participants were part of a group of 260 cleaners, kitchen staff, and clerical workers enrolled from 39 Dutch hospitals between August 1987 and January 1989 before they became pregnant. One hundred and twenty eight of these women were eligible for study because they became pregnant, they worked at least six weeks during pregnancy, and information on work aspects during pregnancy and pregnancy outcome was complete. Work with a high intensity score, and to a less extent work with a high fatigue score, had the most outstanding effect (up to 18 d shorter) on pregnancy duration when the work speed was high. None of the studied aspects of occupational physical activity showed a relevant influence on birthweight when adjusted for pregnancy duration. CONCLUSIONS: This study indicates that the levels of occupational physical load found in the work of nonmedical hospital staff, especially when combined with high work speed, can lead to a shorter pregnancy period. PMID- 7481608 TI - Changes in occupational physical loading during the lifetimes of Finnish men. AB - OBJECTIVES: The aims of this study were to compare summary measures for occupational physical loading based on different periods of work history and to describe how loading exposures change with age and vary by age group. METHODS: Detailed work histories reported by 232 men, aged 35-69 years, were used to compare loading in the current, longest, and heaviest jobs and a lifetime measure and to graph changes in loading over time. RESULTS: The longest job was the best surrogate for lifetime loading (correlation coefficient 0.90). The heaviest work tended to occur in the men's teens and early twenties. For older men, the current or longest job was a poor substitute for the job with the heaviest loading (correlation coefficients 0.24, 0.28). There appeared to be both a cohort and a period effect for sedentary work, lifting and time spent in twisted or bent positions, but not for driving. CONCLUSIONS: If the true risk for back problems is not limited to current activities, exposure misclassification may occur in many studies. PMID- 7481609 TI - Sources of variance in exposure to nonneutral trunk postures in varying work situations. AB - OBJECTIVES: The purpose of this study was to estimate the different components of variance in exposure to nonneutral trunk postures. METHODS: The percentages of worktime spent in trunk flexion and trunk rotation or lateroflexion during 10-min observation periods were used as primary measures in analyses of variance to separate the total variance in its components. For two occupational groups six observation periods were selected per worker, and for four other groups 10 periods were used. Most of the 35 subjects were not restricted to one workplace, and they performed dynamic work while being observed. RESULTS: The variance within workers contributed around 80% to the total variability of exposure to both unfavorable postures. These results indicate that it is difficult to achieve an unbiased estimate of exposure to postural stress for individual workers who perform dynamic tasks in varying work situations. CONCLUSIONS: The results indicate that it is difficult to achieve an unbiased estimate of exposure to postural stress for individual workers who perform dynamic tasks in varying work situations. Measurement strategy for exposure assessment requires many repeated observations per worker. PMID- 7481610 TI - Prevalence of microfungi in Finnish cow barns and some aspects of the occurrence of Wallemia sebi and Fusaria. AB - OBJECTIVES: The occurrence of microfungi in the air and in feeding and bedding materials was studied on 32 Finnish dairy farms. METHODS: Air samples for determining viable and total spore concentrations were collected on membrane filters and with a cascade impactor. Genera of mesophilic, xerophilic, and thermophilic fungi were identified in four culture media. Total spore counts were done with the aid of an epifluorescence microscope. To identify fungal flora in agricultural materials, feeding and bedding material samples were also taken from the farms. RESULTS: The airborne spore concentrations varied for viable mesophilic, xerophilic, and thermophilic fungi from 10(1) to 10(7) colony-forming units per cubic meter, and for total spores from 10(5) to 10(7) spores per cubic meter. Aspergillus, Penicillium, Cladosporium, Absidia species, Wallemia sebi and yeasts were the predominant fungi in the air, as well as in the material samples. CONCLUSIONS: In general, the airborne spore concentrations were high although the variation in the concentrations of different fungal groups was large between the farms. Along with using new growth media, two fungi whose prevalence was earlier poorly known in Finland were detected. W sebi proved to be the most abundant xerophilic fungi in the air and hay samples, while Fusarium spp were very common in grain and straw but rare in air. PMID- 7481611 TI - IARC evaluates wood dust and formaldehyde. International Agency for Research on Cancer. PMID- 7481612 TI - From research to prevention-the 50th anniversary congress of the Finnish Institute of Occupational Health. PMID- 7481613 TI - Dioxins--a health hazard? PMID- 7481614 TI - [Are skin tumors on the increase?]. AB - The skin is the most common site of malignancy. Epithelial tumors, i.e., basalioma and squamous-cell carcinoma, are among the most frequent skin tumors but have a good prognosis if detected early. Prognosis of metastatic melanoma however is bad. Due to increased UV-exposure the frequency of all three tumors has much increased in recent years. Cutaneous lymphomas and kaposi sarcoma are rare skin tumors. The latter is nowadays of increased interest because of its association with HIV infection. An overview of the current epidemiologic data on malignant skin tumors is presented. PMID- 7481615 TI - [Drug-induced neurological complications in a hospital cohort]. AB - Among the patients admitted to our clinic between 1988-1994, we selected 109 with a neurologic disorder induced by medication. We registered 34 cases of drug induced headache, 33 of extrapyramidal syndrome, 10 of polyneuropathy, and 5 of myopathy. 12 patients presented neurologic side effects of antiepileptic drugs, and 15 were admitted for epileptic seizures or neuropsychological alterations induced by drugs. The purpose of this article is to review the clinical symptoms of these types of neurologic disorders. It also discusses the drug-induced neurologic complications encountered in general practice. Therapeutic advice, and particularly the choice of psychotropic drugs for epileptic or elderly patients, is proposed. PMID- 7481616 TI - [Hemoperitoneum due to coitus without vaginal lesions]. AB - A 27-year-old female patient presented in our clinic 14 hours after onset of acute lower abdominal pain during sexual intercourse. Sonography showed ascites around the uterus and structures suspected to be clotted blood. Diagnostic laparoscopy showed in the abdomen about 400 ml of clotted blood: sources of bleeding were small injuries of the left sacro-uterine ligament. Therapy was not necessary, except careful rinsing since hemostasis was already complete. It is important to consider abdominal injuries in cases of abdominal pain after sexual intercourse, even if vulvar or vaginal injuries are absent. PMID- 7481617 TI - [How do melanoma cells circumvent the immune response? Consequences for future therapy attempts and current biopsy processing]. AB - Malignant melanoma can be recognized by the immune system via HLA-associated tumor-associated antigens. Nevertheless, melanoma cells circumvent immune response in most cases. The mechanisms which may be responsible are defects in the expression of HLA-class I molecules, release of soluble adhesion molecules, and secretion of immunosuppressive cytokines such as interleukin-10. Using modern techniques the individual melanoma cells can be cultured and characterized by immunological and molecular biological methods. An exact characterization of the individual tumors will result in specific immunotherapy for the individual patient. PMID- 7481619 TI - [Scientific raisins from 125 years SMW (Swiss Medical Weekly). Biermer's anemia and hypothyroidism. 1940]. PMID- 7481620 TI - [The extent of transfusion-associated HIV infections in Switzerland until 1994: an updated estimate]. AB - OBJECTIVE: The first report on transfusion-associated HIV infections was published in the USA in 1982. The first case reports in Switzerland were published in 1986. So far there has never been a methodologically sound answer to the question of how many persons were infected with HIV by receiving transfusions in Switzerland before the introduction of universal HIV blood donor screening. METHODS: The following available data sources were analyzed simultaneously: firstly, the results of the look-back study conducted in 1993, secondly, the reports of HIV infections and AIDS cases in the national surveillance system, and, thirdly, the claims for compensation for HIV-infected transfusion recipients and hemophiliacs. Two methodologically different and independent estimates were obtained. Firstly, the coverage of the look-back study was estimated, which made it possible to calculate the total number of documentable transfusion-associated HIV infections in Switzerland. Secondly, matching was performed on the cases in the look-back study and the reports in the national surveillance system. Applying formulas of capture-recapture designs provided a second estimate of the total number of documentable transfusion-associated HIV infections. The claims for compensation were used to corroborate the estimates obtained. RESULTS: The two methods produced almost identical figures which were corroborated by the number of claims for compensation. It is therefore estimated that 80 to 100 persons in Switzerland may have been diagnosed as having HIV infection because of transfusions in Switzerland in the years after 1980. The last five known infections occurred in 1986 (four) and, after termination of the look-back study, in 1994 (one). However, the estimate of 80 to 100 does not include individuals who were infected before 1986 and died soon--within weeks or a few months--after the transfusion without diagnosis of HIV infection being possible. CONCLUSION: This estimate of the total number of transfusion-associated HIV infections in Switzerland is approximately half earlier published ones. In addition, the present study will probably reduce the remaining uncertainties about the size of these iatrogenic HIV infections in the 1980s. PMID- 7481618 TI - [Fasciola hepatica--a unusual cause of acute cholecystitis with cholestatic jaundice]. AB - A 29-year-old patient was admitted with acute abdomen in the 17th week of pregnancy. History revealed two episodes of colic in the right hypochondriac area during the previous six months, but no other abdominal complaints. Clinically the picture was that of acute cholecystitis. Laboratory findings included an elevated white cell count, a slight elevation of serum transaminases and a marked increase of serum alkaline phosphatase and bilirubin. Echographically there were dilated intra- and extrahepatic bile ducts containing two hyperechogenic elements without casting an acoustic shadow. A hydrops of the gallbladder with sludge and a thickening of the wall could also been seen. Because of pregnancy an ERCP could not be performed due to the need for X-ray, so we had to resort to open surgery. Under tocolytic and antibiotic shielding we carried out open cholecystectomy and choledochoscopic exploration of the common bile duct. Using a Fogarty balloon catheter we extracted two live, adult liver flukes and placed a T-tube in the duct. Because of positive fecal probes for fasciola eggs the T-tube had to be left in place until childbirth. Afterwards we performed a pre-cut-papillotomy by ERCP and took the T-tube out, having confirmed a clear duct on a T-tube cholangiogram. With negative fecal probes and the eosinophilia on the white cell count returning to normal, we decided against the planned chemotherapy and assumed self-healing of the disease. The patient has been well since.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7481621 TI - [Imported dengue fever in Switzerland--serological evidence for a hitherto unexpectedly high prevalence]. AB - Dengue fever is probably the most important arthropod-borne viral infection worldwide. The World Health Organization estimates an incidence of 100 million cases per year. Dengue fever is of clinical importance mainly in tropical and subtropical zones, but the increasing number of travellers visiting these endemic areas means that physicians in temperate zones will meet this tropical infectious disease more frequently. To estimate the frequency of dengue fever in Switzerland, we retrospectively analyzed a cohort of patients in whom clinicians wished to rule out a case of malaria on the assumption that some of these patients may have been exposed to dengue virus while travelling in tropical and subtropical areas. Dengue was diagnosed by serology on contemporary stored serum samples with a commercially available assay that has been evaluated before. During the study period (1.1.93 to 1.7.94) a malaria diagnosis was requested by the physicians in our center in 87 cases and malaria was diagnosed in 10 (11.5%). In 38 of these 87 cases a stored serum sample was available for dengue serology. All of these patients had visited an area where not only malaria but dengue virus was endemic. In 7 of these 38 cases (18%) malaria was diagnosed. In 3 patients (8%) the dengue serology (IgM and IgG) was indicative of acute dengue fever. All 3 cases had clinical findings compatible with the disease. In 4 more patients (10%) anti-dengue IgG antibodies were found. All of these 4 patients had previous exposure to dengue virus but no acute clinical illness.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7481622 TI - [Malfunction of a mitral bioprosthesis, hemolysis and acute renal insufficiency]. AB - Intravascular hemolysis is a rare complication following valvular replacement, particularly with bioprostheses. It is generally secondary to prosthetic malfunction and is the cause of anemia. We report a rare case of acute renal insufficiency and severe hemolysis caused by malfunction of a bioprosthesis in the mitral position. PMID- 7481623 TI - [Epidemiology of antibiotic resistance: implications for empirical treatment in intensive care]. AB - The adequacy of initial antibiotic therapy is an important prognostic factor in severe infections. Concerning nosocomial infections, the selection of appropriate empirical therapy should take into account the incidence of offending pathogens within a specific unit. The changing trends in the hospital's microbial resistance patterns should be known to the physicians. The bacteria involved and the susceptibility testing vary widely among institutions and among countries. Many risk factors for acquisition of resistant pathogens have been identified. The duration of stay in hospital, previous colonization, and antibiotic treatment are the most frequently cited risk factors. When P. aeruginosa is suspected, an ureido-penicillin/aminoglycoside combination is usually effective. However, in some units with high levels of resistance, the beta-lactam should be ceftazidime or imipenem. When enterobacteria are suspected, a third generation cephalosporin, alone or in combination with an amino-glycoside or a broad spectrum penicillin associated with a beta-lactamase inhibitor is appropriate. Early nosocomial staphylococcal infections are treated with nafcillin or oxacillin, alone or in combination with an aminoglycoside. In units with a high rate of MRSA, the initial antibiotic therapy should include a glycopeptide. PMID- 7481624 TI - [Treatment of constipation in pregnant women. A multicenter study in a gynecological practice]. AB - Constipation is a common problem during pregnancy. It is essential to relieve the condition without putting the developing child at risk or further stressing the mother. The fact that lactulose is not absorbed in the small intestine is of great significance when treating constipation during pregnancy, as it presents no threat to the foetus. Lactulose also does not appear in breast milk so that the treatment can also be continued during nursing. The present study was an open base-line controlled study to show the efficacy, safety and acceptability of dry lactulose (Duphalac Dry) in pregnant women suffering from chronic or occasional constipation. It was a multi-center study in 12 gynecological practices. 62 women aged between 19 and 40 years were treated during 4 weeks with lactulose. 34% had chronic constipation and the remaining 66% constipation due to the pregnancy. The main criteria for efficacy were frequency of defecation and consistency of stools. There was overall judgement of the efficacy by patients and doctors. Safety was assessed by the number of reported side-effects and compared to the reported concomitant symptoms before treatment. Here also, the patients and their doctors gave a global impression. In addition, patients rated the product flavor and administration methods. The frequency of defecation was significantly increased after 1 week (4.0 vs. 2.5 [median per week], p < 0.001) and normalized after 2 weeks (6 stools per week). Also, the consistency of the stools was normalized during the treatment. The global assessment of efficacy by the treating physicians was good or very good in 84% of the cases and by the patients in 81%.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7481625 TI - [Scientific raisins from 125 years SMW (Swiss Medical Weekly. On the familial occurrence of intersexuality and adrenal gland hyperplasia. 1941]. PMID- 7481626 TI - [Tuberculosis: quo vadis? Spotlight on history and current aspects]. AB - In short review some milestones of the history of tuberculosis research are outlined. Perspectives on the past, present and the future of tuberculosis research are linked to these historical facts. Within this context the contribution of members of the Medical Faculty at the University of Berne such as L. Lichtheim, Th. Langhans and E. A. E. Klebs receive particular consideration in connection with the 125th anniversary issue of the "Schweizerische Medizinische Wochenschrift" which was founded in 1871 by Klebs. PMID- 7481627 TI - [Ludwig Lichtheim and Robert Koch. How the discovery of the tubercle bacillus in Switzerland was made known (1882/1883)]. AB - The discovery of the tubercle bacillus by Robert Koch in March 1882 was immediately reported by the medical press, including that of Switzerland. Among the clinicians most active in introducing the new views and techniques to Swiss medical practitioners was Ludwig Lichtheim (1845-1928), professor of internal medicine in Berne (1879-1888), who had been acquainted with Koch since their time in Breslau. Lichtheim's successful efforts are illustrated by quotations from his (unpublished) memoirs. PMID- 7481628 TI - [Current developments in Mycobacteria diagnosis]. AB - The reemergence of tuberculosis in the industrialized countries has hastened the development of new laboratory techniques. Hence, well-known shortcomings of traditional techniques such as the lack of a rapid and specific detection system, the delayed availability of species identification and drug susceptibility results, and the lack of a reliable method for determining strain identity for epidemiological purposes, have become immediate targets for implementing molecular biology techniques. In particular, nucleic acid amplification techniques, restriction fragment-length polymorphism and single-strand conformation polymorphism analyses have dramatically improved diagnostic timeliness and accuracy of mycobacteriology laboratory results. Our paper reviews recent developments and comments on the diagnostic applications of the new tools as compared to traditional techniques. PMID- 7481629 TI - [Laparoscopic adrenalectomy in pheochromocytoma]. AB - Standard adrenalectomy for pheochromocytoma was until recently performed via the transperitoneal open approach. The disadvantage of the open procedure is a more painful and longer recovery. With good surgical experience in laparoscopic techniques and in endocrine surgery, laparoscopic adrenalectomy can be performed safely without complications. The advantage of the laparoscopic technique is excellent vision of the operative field with magnification even of very small vessels. The first two cases of laparoscopic right adrenalectomy at our institution are described. PMID- 7481630 TI - [Scientific raisins from 125 years SMW (Swiss Medical Weekly). Clinico bacteriological studies on sulfonamide resistance of pneumococci. 1942]. PMID- 7481631 TI - [Cardiac rehabilitation--how and when?]. PMID- 7481632 TI - [Current status of cardiovascular rehabilitation and projections for 2010: results of a survey in the Vaud canton]. AB - Compared to the situation in other European countries and the United States, Switzerland chiefly offers inpatient cardiac rehabilitation programmes (CRP). In the opinion of specialists and medical literature, we can expect an increased number of patients with cardiovascular diseases in the next 15 years, as well as a widening of indications for CRP among older patients and those with advanced cardiac conditions. We conducted an inquiry among professionals practising or prescribing CRP in order to estimate population needs and coverage for CRP and to identify the factors that will determine its future. Based on various sources of information, we estimated the incidence of coronary events and demand for CRP in the Canton of Vaud. Cardiac rehabilitation centers provided their occupation statistics for 1993. Simulations were carried out based on the opinion of specialists and expected demographic trends for the next 15 years. In recent years, 600 to 700 patients have been annually enrolled in CRP. Among patients admitted to acute care hospitals after myocardial infarction, 65% of those under 65 years of age and 14% of those between 65 and 74 entered CRP programmes. Most of them followed inpatient programmes in two private clinics which currently cover CRP needs with a capacity of 1500 patients a year. If we expect a 20% increase in the rate of coronary events over the next 15 years, due to demographic trends, without a change in the incidence of coronary disease, the present resources in residential CRP will remain sufficient. However, we can expect that part of the patient surplus will enter outpatient CRP programmes. PMID- 7481633 TI - [Detection of alcoholism in the medical office: applicability of the CAGE questionnaire by the practicing physician. Group of Medical Practitioners PMU]. AB - INTRODUCTION: The general practitioner (GP) plays a very important role in early detection of alcoholism. Clinical evaluation is often the only method used, although it has been suggested that any systematic patient history should include the CAGE test. We compare the effectiveness of these two approaches and attempt to determine the applicability of the CAGE test in a general practitioner's usual practice. METHOD: 12 GPs took part in this study during 6 months. They looked for possible alcohol abuse in each new patient by a standard patient history and clinical examination. Patients were randomized into 2 groups, one of which was given the CAGE test and the other not. For each patient in the CAGE group the applicability of the test was quantified by the GP. RESULTS: 416 patients were included; 214 were randomized into the "CAGE group" and 202 into the control group. On a clinical basis, 15 patients in the control group and 16 in the "CAGE group" (14 men, 2 women) were suspected of alcohol abuse. The CAGE test was positive in 15 patients (7%); among these, 6 were not suspect on a clinical basis. In patients aged 18-34, the detection rate of alcohol-related problems more than doubled when the CAGE test was used. The age of the patients influenced performance of the CAGE test and clinical evaluation. Only 2% of women had a positive CAGE test. Administration of the CAGE test was considered easy in 112 patients and average to difficult in 50, while the test was inapplicable with 52 patients. The latter proportion was higher than that observed in institutions (hospitals, outpatient departments) of the same region. Applicability was influenced neither by the sex nor the age of the patients, but varied greatly according to the physician (from 38% to 100%). CONCLUSION: The CAGE test increases the number of patients detected with alcohol problems by 37% and seems to be especially useful when administered to young people. The number of women with alcohol problems is probably underestimated by both clinical evaluation and the CAGE test. Furthermore, physicians in private practice are more reluctant to use the CAGE test systematically than those in a public institution. PMID- 7481634 TI - [Initial results with laparoscopic fundoplication]. AB - 17 patients underwent laparoscopic Nissen fundoplication for gastroesophageal reflux disease not responding to medical treatment. Conversion to laparotomy was necessarily due to large paraesophageal hiatus hernia (3) or severe peri esophagitis (1) in 4 patients. There was no perioperative morbidity or mortality. At 3 months all patients were symptom-free. Esophageal manometry revealed a rise in mean lower esophageal sphincter pressure from 4 mm Hg preoperatively to 11 mm Hg postoperatively, and lengthening of the high pressure zone from 2.3 to 3.3 cm. One patient needed reoperation one year after fundoplication due to wrap disruption with recurrent reflux disease. Laparoscopic fundoplication involves low perioperative morbidity and shortens the postoperative recovery time in selected patients. PMID- 7481635 TI - [Initial personal experiences with alpha-1-antitrypsin determination in feces]. AB - Fecal alpha-1-antitrypsin is recommended as a marker of enteric protein loss and in patients with Crohn's disease as an index of intestinal inflammatory activity. We describe our experience in 88 patients with chronic diarrhea or suspicion of protein-losing enteropathy. We measured alpha-1-antitrypsin concentration in random stool samples (n = 7), quantitative alpha-1-antitrypsin excretion in a 24 h feces collection (n = 59) and fecal alpha-1-antitrypsin clearance (n = 22). 13 of 88 patients with the following diagnoses had increased values: Crohn's disease (3/9), other inflammatory diseases of the small intestine (3/3, Whipple's disease, eosinophilic gastroenteritis, celiac disease), hypertrophic gastropathy (1/4), infectious diarrhea (2/6), irritable bowel syndrome (2/29), chronic pancreatitis (2/32) and diarrhea of other reasons (0/5). In patients with Crohn's disease, alpha-1-antitrypsin excretion correlated with the clinical disease activity. All 3 patients with other inflammatory diseases of the small intestine showed increased fecal alpha-1-antitrypsin. All but 2 of the 32 patients with diarrhea due to chronic pancreatitis had normal values. Of 29 patients with idiopathic diarrhea, only 2 showed slightly increased fecal alpha-1-antitrypsin. 10 of the 11 patients with increased alpha-1-antitrypsin excretion in 24 h stool collection had normal alpha-1-antitrypsin concentration in random stool samples. Of the 5 patients with increased alpha-1-antitrypsin clearance, 4 also had increased alpha-1-antitrypsin in 24 h stool collection, but only one had increased alpha-1-antitrypsin concentration in random stool sample. Fecal alpha-1 antitrypsin measurement proved helpful in differing between inflammatory and non inflammatory diarrhea.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7481636 TI - [Scientific raisins from 125 years SMW (Swiss Medical Weekly). Myalgia epidemica in Switzerland 1940 (Bornholm disease in Gais and Teufen). 1941]. PMID- 7481637 TI - [Epidemic of type B botulism: Sion, December 1993-January 1994]. AB - 12 cases of food-borne botulism were registered in Sion, Switzerland, between 31 December, 1993 and 12 January, 1994. A type B toxin was isolated from the serum of one patient and from the incriminated ham. Clinical data of 10 male patients aged 21 to 54 years and some epidemiologic data are reported. The clinical course was mild to moderate with predominant autonomic and gastro-intestinal symptoms and signs: blurred vision (10 patients of 10), dry mouth with dysphagia (9/10), asthenia (7/10), diarrhea and/or constipation (7/10), nausea and vomiting (6/10), abdominal cramps (5/10), impaired sexual function (5/10), dilated pupils (4/10). Some discomfort (mainly blurred vision, asthenia and impaired sexual function) persisted for several months in most patients. Neuromuscular involvement was never the reason for seeking medical assistance and had often disappeared at the time of the first visit. Two patients were hospitalized, one for transient ileus of unknown origin and the second (first suspected case) for monitoring and infusion of trivalent equine botulinum antitoxin. This treatment was administered on day eight after intoxication and had no effect on this patient's outcome when compared with others. No patient died. Epidemiology, diagnosis, treatment and prognosis of botulism are discussed. PMID- 7481638 TI - [Postoperative work incapacity, type of work resumed and residual pain following surgical treatment of lumbar herniated disk: prognostic factors]. AB - The outcome in 258 patients 3 to 4 years after operation for lumbar disc herniation was studied. The median duration of postoperative work incapacity was 3.5 months; 202 (78%) patients resumed full time work in their previous job; 16 (6%) did not resume work due to lumbar pathology; 193 (75%) were free of pain. Predictive factors for a good professional outcome were a preoperative work incapacity inferior to 4 weeks and, for men only, operation at the L5-S1 level and the absence of daily lifting of heavy weights. PMID- 7481639 TI - [Risk in major hepatectomy. A consecutive series of 113 extensive hepatectomies]. AB - Up to now, liver resections have been the initial treatment of almost all cancers and benign tumors limited to a liver lobe. This retrospective review assesses the results of a consecutive series of 113 major elective hepatic resections during a ten-year period. Major hepatectomy was defined by the resection of at least 3 Couinaud segments. Mean age was 52 years (20 to 79 years). There were 62 women and 51 men. 35 resections were performed for colorectal metastases, 22 for a benign tumor, 20 for non-colorectal metastases, 11 for hydatid disease, 10 for hepatocarcinoma, 7 for cholangiocarcinoma and 8 for other indications. The resections performed were 86 right hepatectomies with 18 extended right hepatectomies, 24 left hepatectomies with 4 extended left hepatectomies and 3 trisegmentectomies. Total vascular exclusion was used in 22 patients (19%). Mortality rate was zero. Significant morbidity was encountered in 24 patients (21%). These results suggest that the mortality rate may be independent of the extent of liver resection, provided that hepatic function is normal and preoperative selection adequate. With improving surgical management and techniques, and the use of intra-operative sonography, extensive liver surgery can now be performed with a very low mortality rate. PMID- 7481640 TI - [Does laparoscopic colonic carcinoma surgery satisfy the radicality criteria of open surgery?]. AB - Minimal invasive surgery is applicable to almost all colorectal operations, with major benefit for the patient. Technically even cancer operations can be performed. However, in laparoscopic assisted colorectal surgery the question of radicality outweighs that of performability and patient comfort. From a prospective series of 88 laparoscopic colorectal operations, 36 were for carcinoma. 34 patients who underwent conventional surgery were matched with regard to age, sex, type of operation and tumor stage (TNM, grading) to compare the two techniques. The two interventions followed exactly the same guidelines. We compared the length of the fixed specimens, the number of resected lymph nodes and the need for blood transfusions. Postoperative complications were noted and follow-up was 3-12 months. The data obtained showed no difference between the two treatment groups, with a slight trend towards laparoscopic surgery with regard to number of resected lymph nodes. A portside metastasis was observed 9 months after an initial tumor stage T4N1M1. The close relation between the portside and infiltration of the adenocarcinoma into the abdominal wall together with a drain placed through this whole was suspected to be the cause of this complication. We conclude that laparoscopic assisted colorectal surgery for cancer is equal to open operation regarding radicality of resection. Long term results are mandatory to determine the value of minimal invasive surgery in the field of oncology. PMID- 7481641 TI - [How reliable is the measurement of colonic transit time using a marker technique?]. AB - Radioopaque markers are well established tools to determine segmental and total colonic transit time. However, since no data are available on intraindividual reproducibility, in this study reproducibility was assessed in 14 health male and 12 healthy female volunteers. The mean segmental colonic transit and total colonic transit time were not significantly different for the two measurements. Colonic transit time was consistently shorter in males than in females. The menstrual phase did not show a significant influence in the latter. The intraindividual variations of the colonic transit times were acceptable with a reproducibility coefficient of 20 h. PMID- 7481643 TI - General medicine and geriatrics, where is the difference? The example of infective disease. AB - Geriatric medicine differs from internal medicine not in quality, but rather in the probability structures of diagnosis and outcome, presentation of illness and the need for explicit determination of aim of intervention. Older people need more investigation than the young if comparable levels of diagnostic accuracy are to be obtained. In view of the progressive loss of adaptability with its loss of sufficient functional and social reserves, the old more often need a formal program of rehabilitation than younger patients. Infectious diseases exemplify this difference. Age associated impairment of immunity, such as lower T-cell activity and IL-2 production, are associated with an increasing lethality of infections, and permit the reactivation of latent infections. The loss of adaptability has its impacts on mental functioning by increasing the risk of delirium as a consequence of infectious diseases or drug side effects. The cryptic presentation of illness makes even severe infections such as endocarditis, peritonitis or tuberculosis difficult to diagnose. The traditional exclusion of older people from research studies contributed to the still prevailing underestimation of the complexity and need of adequate therapy in geriatric patients. PMID- 7481642 TI - [Scientific raisins from 125 years SMW (Swiaa Medical Weekly). 2nd international medical week dedicated in Switzerland. Luzern, 31 August-5 September 1936. Fibroplastic parietal endocarditis with eosinophilia. An unusual disease. 1936]. PMID- 7481644 TI - [Does serum HCV-RNA-positive hepatitis C differ from serum HCV-RNA-negative hepatitis C?]. AB - Viral RNA is detectable in the serum of the majority but not all patients with chronic hepatitis C. Whether the viremic form differs from the non-viremic form of the disease is unknown. We therefore compared histology (modified Knodell score) and liver function (conventional liver function tests, galactose elimination capacity and aminopyrin breath test) of viremic (n = 45) and non viremic (n = 37) patients with chronic hepatitis C. Neither the total histologic score, nor any of the individual histologic parameters assessed differed significantly in serum HCV RNA positive and negative patients. Compared to non viremic subjects, patients with detectable HCV RNA in serum had slightly higher transaminases (p = ns), lower serum albumin (p < 0.05) and decreased galactose elimination capacity (p < 0.05). This trend towards more severe functional hepatic impairment in serum HCV RNA positive patients persisted when cirrhotics and non-cirrhotics were analyzed separately; it fell just short of reaching statistical significance, however, most probably due to the small number of subjects per patient group. The median age of serum HCV RNA positive cirrhotics was 17 years, and that of serum HCV RNA negative cirrhotics 22 years higher than that of the respective non-cirrhotics. Biochemical (transaminases) and histological (intralobular and piece-meal necrosis) markers of disease activity, as well as metabolic functional reserve (aminopyrin breath test, galactose elimination capacity) and histologic severity of fibrosis correlated only loosely (Rs = 0.24-0.56), albeit significantly (p < 0.05).(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7481645 TI - [Gluten-sensitive enteropathy with intestinal T-cell lymphoma: an unusual cause of in disabling osteomalacia]. AB - We describe a 64-year-old woman with a malignant intestinal T-cell lymphoma who presented four years later with disabling osteomalacia and secondary hyperparathyroidism due to malabsorption. Only two years later, when the patient had developed fatty stools, flatulence and weight loss, diagnosis of gluten sensitive enteropathy (GSE) was confirmed by small-intestine biopsy. This case report illustrates that in adults GSE can be oligosymptomatic for long periods. In cases of osteomalacia or rare intestinal T-cell lymphoma a detailed history of bowel movements, inspection of stools, quantification of fat excretion in stools and laboratory tests for malabsorption are recommended. Positive antibodies against gliadin, endomysium and reticulin may support the diagnosis of GSE. However, intestinal biopsy is necessary to verify the presence of GSE. In view of the unspecific histological changes, a follow-up biopsy is recommended in oligosymptomatic cases. Serial measurements of antibodies allow supervision of compliance for a diet strictly free of gluten. In addition, lactose containing milk products need to be restricted initially because of secondary lactase deficiency. PMID- 7481646 TI - [Delayed allergic reaction to Chlorambucil (Leukeran). Case report and literature review]. AB - A 63-year-old man developed a mild hemorrhagic diathesis which led to the diagnosis of chronic lymphatic leukemia. Treatment with prednisone and chlorambucil was initiated. During the first 10 day-cycle (10 mg chlorambucil and 100 mg prednisone) no side effects were noted. On the 8th day of second cycle (10 mg chlorambucil and 25 mg prednisone) the patient noticed fever, tiredness, myalgia, pruritus and erythema on the skin. The third cycle (10 mg chlorambucil alone) hat to be stopped on the second day due to the development of myalgia, generalized erythroderma with exfoliation and edema of the face and arms. In the patch-test with chlorambucil, a strongly positive (histologically verified) allergic reaction was noted. A lymphocyte stimulation test (detecting in vitro lymphocytes sensitized to chlorambucil) was also positive. These findings, together with the marked clinical reaction to chlorambucil alone, led to the diagnosis of a delayed hypersensitivity reaction to chlorambucil. Such well documented allergic reactions to chlorambucil are very rarely described in the literature. Up to now only 1 case of immune hemolytic anemia, 1 case of Lyell syndrome, 2 cases of delayed hypersensitivity reactions and a few cases of less well documented type I reactions have been described. In all these cases no crossreactivity with other alkylating agents occurred, and hence all patients could be successfully treated with cyclophosphamide. PMID- 7481647 TI - [Dysphagia, cavernous changes of the lungs and subarachnoid hemorrhage following kidney transplantation]. AB - A 54-year-old patient with polycystic kidney disease developed relapsing urinary tract infections after renal transplantation. In the post-transplantation period he underwent two rejection episodes. The general condition worsened without obvious reason. Six months after transplantation the patient was hospitalized due to gram-negative sepsis. Despite adequate antimicrobial treatment he continued to lose weight and complained of dysphagia. Chest X-ray and computer tomogram of the thorax showed multiple cavitating infiltrations of the lung. A few days later the patient died from an extended subarachnoidal hemorrhage. PMID- 7481648 TI - [Scientific raisins from 125 years SMW (Swiss Medical Weekly). Picture and early course of tuberculosis in patients from concentration camps.1947]. PMID- 7481653 TI - [Cardiac pacemaker for the treatment of obstructive hypertrophic cardiomyopathy, a concept which is gaining ground]. AB - Pacing reduces the sub-aortic pressure gradient in hypertrophic obstructive cardiomyopathy. This phenomenon was described long ago but only recently, with the development of sophisticated pacemakers, has it become possible to use pacemakers for the long term treatment of this disease. The acute hemodynamic improvement with pacemaker treatment is well documented. The reason for this acute and chronic improvement is of multiple origin. Pacing modulates the activation sequence of the left ventricle, reduces contractility, reduces subaortic flow velocities, influences mitral valve movement and leads to remodeling of the ventricle in the long term. All these factors together explain the clinical and hemodynamic improvement in patients suffering from hypertrophic obstructive cardiomyopathy. PMID- 7481649 TI - [Patient management following cardiac transplantation]. AB - Prognosis of heart transplant patients has dramatically improved in the last decade thanks to the use of cyclosporine as immunosuppressive treatment. Optimal follow-up and early recognition of complications and side effects of treatment lead to longer survival and marked improvement of these patients' quality of life. However, the incidence of complications remains high and their management calls for particular care. We review our experience of heart transplantation in Lausanne and describe in detail the results and complications such as rejection, transplant coronary artery disease, hypertension, renal failure, metabolic disorders, cancer and conduction problems. Cooperation of the medical team (cardiologist, surgeon, anesthetist, general practitioner) is essential to long term success of the transplant program. PMID- 7481650 TI - [Coronary endothelial dysfunction and graft atheromatosis following heart transplantation]. AB - Graft atheromatosis is the most important limiting factor on long-term survival after heart transplantation. Histologically it involves so-called myointimal proliferation occurring in either circumscribed or diffuse form. Endothelial dysfunction with impaired release of nitric oxide represents an early stage of graft atheromatosis. Progression of the disease typically leads to a diffuse narrowing of the coronary tree; however, focal stenoses may also occur. Endothelial dysfunction results in a decrease in physiological coronary flow reserve during exercise, whereas pharmacological flow reserve after papaverine or adenosine administration is maintained. This functional disturbance can be enhanced by transplantation-related (e.g., vascular graft rejections, cytomegalovirus infections, etc.) as well as by cardiovascular risk factors (e.g., hypercholesterolemia, hypertension). The occurrence of endothelial dysfunction and graft atheromatosis may be delayed, although probably not prevented, by elimination of risk factors and optimization of immunosuppressive treatment. Preliminary data suggest that long-term administration of the calcium antagonist diltiazem may have a protective effect. PMID- 7481651 TI - [Hypertrophic cardiomyopathy: spontaneous course]. AB - Hypertrophic cardiomyopathy is a relatively rare (prevalence approximately 0.2%), primary myocardial disorder with an autosomal pattern of inheritance, characterized by mostly asymmetric left ventricular hypertrophy with myocyte and myofibrillar disarray. To date, about 34 mutations of the beta-cardiac myosin heavy chain gene have been described and shown to have prognostic implications. The disease has an annual mortality rate of 3%, related to both sudden cardiac death and progressive systolic dysfunction. Not only diastolic but also progressive systolic dysfunction with cavity dilatation occurs in a minority of patients with severe hypertrophy during the long-term course. Sudden death often occurs in young, asymptomatic or mildly symptomatic patients. The degree of hypertrophy and the presence of a pressure gradient are of little prognostic significance. Nonsustained ventricular tachycardia is associated with a poor prognosis in the presence of a history of syncope. PMID- 7481652 TI - [Hypertrophic obstructive cardiomyopathy: surgical treatment]. AB - Hypertrophic obstructive cardiomyopathy (HOCM) is an autosomal, dominant inherited disease of the myocardium which leads slowly to increasing subvalvular, septal and left ventricular hypertrophy and deterioration of systolic and diastolic left ventricular compliance. Difficult molecular-genetic investigations localized genetic defects on different chromosomes. The disease is pathological anatomically characterized by asymmetric subvalvular (aortic) septal hypertrophy and left ventricular outflow tract obstruction resulting in additional left ventricular hypertrophy and dysfunction. Histologically the myocytes are hypertrophied, exhibit atypical branching (disarray), and there is a high amount of interstitial connective tissue. In our biopsy material (from myectomies) dysplasia could be detected in more than 30% of dysplastic intramural arteries with partly extential media. These changes may indicate microcirculatory disturbances resulting in arrhythmias, syncopes, sudden death, and anginal pain on the basis of microcirculatory disturbances and scar development. Today the discussion of DDD-pacemaker therapy has resumed, but one must wait for definite results, especially in patients in whom surgical treatment seems to be the best choice. The indication for surgical treatment, which usually is transaortic subvalvular myectomy (Morrow) and modifications, is very restrictive. Only patients in clinical degree III (NYHA) after long-term medical treatment are candidates for surgery. In some mainly younger patients the indication in lower clinical degrees was accepted because of a family history with sudden death and personal experience of syncope, life-threatening tachycardia, or after resuscitation. In the period 1963 to 1994 466 patients were operated upon. The mean age was 44.9 years (range 3 months to 82 years). Total early mortality was 4.9% (n = 12).(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7481654 TI - [Scientific raisins from 125 years SMW (Swiss Medical Weekly). On Parpanit, a new agents which modifies extrapyramidal motor disorders. 1946]. PMID- 7481655 TI - [Health care economics, uncertainty and physician-induced demand]. AB - The health care market is a very particular one that is mainly characterized by the absence of information and transparency at every level, particularly between the physician-supplier and the patient-consumer. On this market it is up to the physician to evaluate and define the patient's needs and to decide which are the most effective goods for the patient. The determinants of medical prescription are not only related to the health status of the patient, but also to the payment system (fee for services, salary), to physician density, professional uncertainty, the role and status of the physician in his profession, the legal framework which rules the medical profession, and also the information level of the patient. Agency relationship and professional uncertainty are the most relevant determinants of supplier-induced demand. Professional uncertainty inherent in the practice of a stochastic art such as medicine will "always" give an ethical justification for supplier-induced demand or for the pursuit of "maximal" and/or "defensive" care when market competition is perceived by the physician as a threat to his/her income or employment. Time is ripe for consumers and physicians empowerment in the aim to promote better self-management of health and more thoughtful access to care (for consumers) and more evidences based medicine for physicians. PMID- 7481656 TI - [Permanent junctional reciprocating tachycardia: a little-known clinical entity curable with radiofrequency ablation]. AB - We report our experience of 5 patients with the permanent form of junctional reciprocating tachycardia (PJRT), a rare form of supraventricular arrhythmia. PJRT was discovered at a mean age of 31 years (8-60 years) and the mean duration of tachycardia was 13 years (1-40 years). 4 patients had nearly incessant tachycardia and one had paroxysmal attacks. Heart rate varied between 100 and 190 beats/minute and the minimal heart rate was on average 114 beats/minute. Four patients had palpitations, 2 developed tachycardia-induced cardiomyopathy, reversible after control of the arrhythmia, and 4 had asymptomatic episodes of PJRT. ECG showed in all cases a narrow-complex tachycardia with inverted P waves in inferior leads and RP interval greater than PR. All patients presented a posteroseptal accessory pathway. 4 patients received different antiarrhythmic drugs with only partially effective results. Radiofrequency catheter ablation of the accessory pathway was performed in all patients and was successful in 4, who remained free of recurrence after a mean follow-up of 26.5 months (4-37 months). The procedure was partially successful in the 5th patient, who is now asymptomatic under sotalol. Radiofrequency catheter ablation is therefore the treatment of choice of PJRT, a rare arrhythmia which should nevertheless be known in order to treat the patient correctly and avoid progression to cardiac failure, which is not always completely reversible. PMID- 7481657 TI - [Human echinococcosis in Switzerland, 1984-1992]. AB - In a retrospective study (1984-1992), new cases of human echinococcosis were registered in Switzerland based on information obtained from (a) questionnaires sent to 294 of the 300 acute hospitals in all parts of the country and to 17 institutes of pathology (268 answers form hospitals: 91%), and (b) from tracing back cases reported under the official notification system since 1 January 1988 by laboratories to the Federal Health Office or recorded at the Institute of Parasitology in Zurich. Cases were regarded as verified if the diagnosis was documented by unequivocal findings (by radiology, ultrasonography, pathomorphology etc. and often by additional detection of anti-Echinococcus antibodies). Patients with antibodies but without reports on further findings were classified as suspected cases. From 1984 to 1992 (9 years), 302 new cases of human echinococcosis were diagnosed in Switzerland and verified in this study, corresponding to an annual average of 34 new cases and a range between 26 and 43 cases. The total number of 302 cases included 228 (75%) of cystic echinococcosis (CE) (Echinococcus granulosus), 65 (22%) of alveolar echinococcosis (AE) (E. multilocularis), and 9 (3%) of non-specified echinococcosis (NSE) (Echinococcus sp.). Among 185 patients with CE and 60 patients with AE and known geographic origin, the ratio of Swiss nationals to foreigners was 25%:75% and 88%:12% respectively. Based on a total population (Swiss nationals and foreigners) of 6.62 million in 1988 and the case numbers of 1984-1992, the following average annual incidence rates per 100,000 inhabitants were calculated: 0.51 for all new cases, 0.38 for CE, 0.11 for AE and 0.01 for NSE. In the 37 years since 1956 there has been steady increase in new cases of CE due to the importation of such cases by foreigners, whereas the case numbers of AE have remained nearly constant with a range between 7 and 10 new cases per year. In our study 258 suspected seropositive patients were registered but not added to the total number of cases, due to the lack of further findings. PMID- 7481658 TI - [Crohn disease in an AIDS patient]. AB - We report on a 32-year-old homosexual AIDS patient with Crohn's disease of the ileocecal region. Radiographically the bowel of this region showed thickening of the wall and moderate narrowing of the lumen. Sonography showed an enlargement of the mesenterial lymph nodes. Due to mechanic ileus, ileocecal resection was performed. Histology of the resected bowel segment revealed extensive inflammatory changes throughout the wall and deep ulcerations, corresponding to changes in Crohn's disease. An underlying infection was histologically excluded. The immunologic aspects of Crohn's disease and of AIDS, and the differential diagnoses of Crohn's disease in AIDS patients, are discussed. PMID- 7481660 TI - [Scientific raisins from 125 year SMW (Swiss Medical Weekly) Q fever (Queensland fever) in Switzerland (endemic pneumonia caused by Rickettsia burneti). 1948]. PMID- 7481659 TI - [Assessment of disease course in chronic rheumatoid arthritis: new quantitative dimensions for clinical practice]. AB - Longterm follow-up of patients with rheumatoid arthritis requires systematic measurement of all relevant disease dimensions. The disease process can be measured reliably with the disease activity score (DAS) which integrates the erythrocyte sedimentation rate and the number of swollen and the number of tender joints out of 28 defined joints. Cumulatively the disease process may result in joint damage which can be measured with a radiological score averaging the level of destruction of 10 key joints. Synovitis and/or joint damage lead to reduced joint mobility and muscle strength which may be quantified using a muscle strength index (MSI) which integrates the information of isometric muscle strength of knee and elbow extension and flexion. The most important outcome dimensions are symptoms and physical functional disability, which can be measured comprehensively with a symptom-oriented rheumatoid arthritis disease activity index (RADAI) and the health assessment questionnaire (HAQ). Reduction of a wide range of information to a few valid indices representing all relevant disease dimensions allows interpretable but comprehensive evaluation of the disease course. PMID- 7481661 TI - [Annual meeting of the Swiss Society of Rheumatology and The Swiss Society of Physical Medicine and Rehabilitation. Loeche-les-Bains, 7-9 September 1995. Abstracts]. PMID- 7481662 TI - [Annual meeting of the Swiss Society of Intensive Medicine and the Swiss Society of Anesthesiology and Resuscitation. Zurich-Irchel, 28-30 September 1995. Abstracts]. PMID- 7481663 TI - [Violence--an element of the psychiatric profession]. AB - In an article on principles it is shown that violence plays a significant part in psychiatry: The patients bring violence into the psychiatric institution as a symptom as well as an origin of their disorders. The institution itself can again not be free of structural violence. Add to this a resonance in the personal aggressive emotions of the professionals. In this way violence becomes an element of the psychiatric profession. In the centre of this appears the specific competence of the psychiatrist to judge about the ability of judgement ("reality control") of other people in an authoritarian manner. This competence which cannot be eliminated from the psychiatric work gives power and has a latent totalitarian aspect. Thus taking over the responsibility to limit the violence that is set free through psychic disorders we have to limit this responsibility in the same time. PMID- 7481664 TI - [Psychotherapy with dyssocial patients]. AB - Psychotherapy of dissocial personalities is, even by psychotherapists, often judged sceptically. As obstacles are most of all mentioned the lack of motivation and the excessive acting out of these patients. Because of the development of the personality of dissocial people motivation in the ordinary sense can not be expected. But this does not mean that therapy is impossible. These patients are often rejected by therapists because they feel injured by the patients' rejection and by their devaluation of the therapeutic attention. We find the same dynamics concerning the excessive acting out or these patients. Problems in countertransference also arise because therapists must reduce their therapeutic ambitions and must be satisfied with improvements which-in their eyes-are negligible. PMID- 7481666 TI - [Psychotherapy with violent neurotic patients]. AB - The paper presents a model of interactional processes for analytical psychotherapy with violent neurotic patients. The starting point is a special situation of transference related to the negative therapeutic reaction: The interaction is reduced to the demand for being satisfied immediately by the therapist. A model of the conditions for the emerging of violent acting out is developed and the structure of transference with violent neurotic patients is exposed. Special difficulties in the managing of transference and the technique of interventions are discussed. PMID- 7481665 TI - [Epidemiology of violence in psychiatry]. AB - An overview of the literature indicates that violence in psychiatry is caused by a minority of patients. Despite the lack of precise definition, between 3 and 10% of patients shows an assaultive behaviour, at least once during the hospitalisation, but, fortunately, injuries are exceptional. The authors present some results of a research project evaluating the frequency and the cost of violence. PMID- 7481667 TI - [Therapeutic reflections on violence in schizophrenic patients]. AB - A model for a psychotherapeutic access to the violence of schizophrenics is presented. The theory bases on three schizophrenic operations: The loss of thinking correlated to the absence of the psychical representative for the act of violence. The splitting of the ego by which the patient knows of his violence but does not remember it. The paradoxon of speaking and not speaking about violence in the same time. The active use of these psychotic mechanism in the therapy and technique is showed by casuistics. Special problems are discussed. PMID- 7481668 TI - [Identity crisis and training of psychiatrists]. AB - A trend towards remedicalization in Psychiatry has become apparent in the 80's and 90's. Most psychiatrists tend however to maintain an eclectic attitude, although a few manifest a clear identity crisis leading sometimes to an hyperidentification with one of the psychiatric theoretical models. The training programs for psychiatric residents should take into consideration two important factors in the building up of the professional identity of the future psychiatrists: the tendency of the residents to identify with the ideology of their teachers and the modelling effect of the different types of clinical activities on the theoretical approach of the clinician. PMID- 7481669 TI - [Quality management in dentistry?]. PMID- 7481671 TI - [Materials and technics in root canal treatments in Switzerland--a determination of their status]. AB - No data is available on either the materials nor techniques used by Swiss clinicians when doing root canal therapy (RCT). Consequently, in January 1993 questionnaires were posted to all dentists registered with the Swiss Dental Association, excluding those known not to practice RCT. Sixteen questions were answered detailing the disinfecting and obturating materials used, which types of files, preparation and obturation techniques were used and which radiographs were taken during RCT. Demographic details were also answered. Of the 3353 questionnaires posted, 2091 (62.4%) were completed and returned. Major results showed that 50.6% of the responding clinicians used AH26 to obturate canals, 45% Endomethasone and 36.6% N2. Canals were disinfected using Ledermix (81.2%), Asphaline (67.6%), Ca(OH)2 (60.2%). Clinicians used H2O2 and NaOCl equally frequently (75%). Hedstroem files, reamers and flexofiles were used by 52%, 29% and 26% of the clinicians, respectively. Canals were obturated by single-cone technique (68%), lateral condensation (34%), or by using only a paste or sealer (23%). 86.4% take a diagnostic radiograph, 69.2% determine working length from radiographs, and 91.5% take a final radiograph as well. Too many Swiss clinicians still use formaldehyde-containing products in root canal therapy coupled with a "single cone" or a "no cone" obturation technique. PMID- 7481672 TI - [The subjective evaluation of oral health in 40- to 69-year-old subjects. A representative survey of 600 persons in German- and French-speaking Switzerland]. AB - Individual oral hygiene, professional dental care and subjective satisfaction with the oral health were investigated in 600 persons representative for the German- and French-speaking population of Switzerland. They cleaned their teeth on average 2.4 x per day, 85% had a "family dentist" whom they had last visited 14.6 months ago and who works to their full satisfaction. On average, every 13.5 month they underwent professional cleaning of their teeth which was performed in 55% by their dentist and in 45% by a dental hygienist. They were well satisfied with the condition of their teeth and gums and even better so with their reconstructions. In 1984 a gap of consumption of toothbrushes and toothpastes had been apparent in over 50-year-old people. Today this is just visible in the over 60 year old. 40 to 60 year old people care for their oral hygiene and regularly go to professional control and cleaning appointments. Only over 60 year old people are consuming slightly less dental services and buying less toothbrushes and toothpastes. Turning points are loss of teeth, retirement, isolation and sharp cuts in income. PMID- 7481673 TI - [The computer-generated occlusion of Cerec-2 inlays and onlays]. AB - The Cerec-2 unit offers three different options to generate the occlusion: anatomically extrapolated ("Extrapolation"), correlated to a wax-up or functionally generated path ("Correlation") and bucco-orally flat ("Linear") as known from Cerec-1. These three computer-generated occlusions were evaluated by measuring the vertical discrepancy and the specific time needed for manufacturing (design, machining, contouring and adjusting the occlusal relation) of inlays and overlays. The occlusion of inlays generated with "Extrapolation" (0.42 +/- 0.16 mm) and "Correlation" (0.80 +/- 0.17 mm) showed significantly (p < 0.001) less vertical discrepancy than "Linear" (2.46 +/- 0.50 mm). The specific production time of inlays differed significantly (p < 0.001) between "Extrapolation" (16.05 +/- 0.80 min), "Linear" (19.25 +/- 1.14 min) and "Correlation" (27.33 +/- 0.75 min). The vertical discrepancies of overlays generated with "Correlation" (1.26 +/- 0.43 mm) were significantly (p < 0.001) lower than with "Extrapolation" (2.46 +/- 0.50 mm) and "Linear" (2.75 +/- 0.57). The specific overlay production time was significantly (p < 0.001) longer with "Correlation" (39.44 +/- 1.36) than with "Linear" (23.9 +/- 1.26 min) and "Extrapolation" (23.71 +/- 1.01 min). Related to the technical status of 10/1994 and as a result of the present evaluation, "Extrapolation" was chosen to be the most practical of the computer generated occlusions evaluated in the present study. PMID- 7481670 TI - [Damage to neighboring teeth during the preparation of proximal cavities. An in vivo study]. AB - It has been shown that iatrogenic preparation damage increases caries progression and the need for restorative therapy of the adjacent tooth. The aim of this study was to quantify the damage of the adjacent tooth using three different preparation techniques: (1) diamond bur alone; (2) diamond bur combined with the EVA system and the Cavishape file; (3) diamond bur combined with an axial margin trimmer. All 136 preparations were performed by dental practitioners in their own practice. Examination of the area directly opposite to the axial box margin revealed significantly less damage (p < 0.01) using additionally the EVA system with the Cavishape file or the axial margin trimmer compared to using only diamond burs. The area opposite between the two box margins, which was in all three methods prepared only by the diamond bur, showed no significant differences between the methods under study. There were only a few teeth (overall 5%) with no damage of the adjacent surface. In order to reduce iatrogenic damage for the adjacent teeth the preparation technique has to be modified. PMID- 7481674 TI - [Impressions and esthetic rehabilitation. The preparatory work, clinical procedures and materials]. PMID- 7481675 TI - [The voltage-dependence of battery-operated light-curing units]. PMID- 7481676 TI - [The therapy of dens invaginatus. A clinical concept for prevention and therapy in teeth with a dens invaginatus]. PMID- 7481677 TI - [The Federation Dentaire Internationale: a modern umbrella organization]. PMID- 7481678 TI - ["To be convincing, dentists must appear creative"]. PMID- 7481679 TI - [Quality management and ISO 9000 certification]. PMID- 7481680 TI - [Patients at risk, social circumstances, medicolegal activities and general dentistry. Interview by Kurt Venner]. PMID- 7481681 TI - [The risk patient in dental practice. A report on the SSO congress of 18-20 May 1995 in Lugano. Societe Suisse d'Odonto-Stomatologie]. PMID- 7481683 TI - [Constructive discussions. A report on 45th Congress of the German Society for Oral and Maxillofacial Surgery in Erfurt]. PMID- 7481682 TI - [CAR 95. A report on the 9th International Symposium CAR 95 (Computer Assisted Radiology) of 21 to 24 June 1995 in Berlin]. PMID- 7481684 TI - [3 symposia on minimally invasive surgery. A report on the 12th International Congress for Oral and Maxillofacial Surgery of 29 June to 2 July 1995 in Budapest]. PMID- 7481685 TI - [The anatomically difficult maxilla--preprosthetic surgery and implants. A report on the 46th annual meeting of the Association for Oral Surgery within the German Society for Dentistry, Oral Medicine and Orthodontics jointly with the German Society for Implantology in Dentistry, Oral Medicine and Orthodontics, Inc. from 25 to 27 May 1995 in Bad Homburg]. PMID- 7481686 TI - ["Cost savings in the area of oral prophylaxis will be expensive"]. PMID- 7481687 TI - [Biocem--a new material for retrograde root filling. The 5-year clinical results]. AB - This paper describes the application of a new material, Biocem, for retrograde root canal filling. A total of 234 roots in 222 teeth were treated during the period 1987-1992. Biocem consists of two components and once mixed, the autopolymerisation results in a three-dimensional structure. The biomechanical and histologic properties and a clinical longterm follow-up confirm the optimal qualities of this material. It fulfills the requirements in regard to biocompatibility, watertight sealing and application. The material proved superior in comparison to conventional materials and ceramics. The evaluation of the results are most encouraging. PMID- 7481688 TI - [Mercury emission measurements in a crematorium. The dentistry aspects]. AB - The amount of mercury released during cremation was calculated in a major Swiss city at two crematoria. A total of 60 mercury "output" calculations were carried out by the Swiss Material Testing Institute (Empa). The amount of mercury initially present ("input") in the dentitions of 54 deceased persons was assessed from their post-mortem dental radiographs and by clinical examination. The correlation between the "input" and the "output" was 0.93, irrespective of the age at death. However, the "input" was calculated to be 1.8 times higher than the "output" for the deceased people with amalgam restorations. In the blind study, the difference was 1.3 times. The main source of mercury was undoubtedly the amalgam restorations. The amount of mercury recorded during the cremation of 88% of the deceased people without amalgam restorations was under the accepted level of 200 micrograms/m3. However, in 3 cases, the amount of mercury was slightly more than 200 micrograms/m3. In contrast, the amount of mercury recorded during the cremation of only 18% of the deceased people with amalgam restorations was less than the accepted level of 200 micrograms/m3. The amount of mercury contamination during cremation as a result of amalgam fillings is so low that no additional preventive measures are required at the crematoria. PMID- 7481689 TI - Collagen metabolism and tooth eruption: the effects of sodium morrhuate infusions on premolar eruption in dogs. AB - To test the essential contribution to tooth eruption of the known high level of collagen metabolism in the periodontal ligament, we have infused the crypts of erupting premolars in dogs with sodium morrhuate, a compound known to reduce production, hydroxyproline content and maturation of collagen. Infusions of sodium morrhuate early or later in eruption for more than half the period of eruption had no effect on the process evaluated radiographically and clinically. These data, considered together with other studies, suggest that collagen metabolism per se plays no essential role in tooth eruption. PMID- 7481690 TI - [Cerec restorations. Cerec inlays and onlays: the clinical results and experiences after 6 years of use in private practice]. PMID- 7481691 TI - [Waste management in the dental office]. PMID- 7481693 TI - [Esthetics in implantology. A report on the scientific meeting of the Societe Suisse d'Implantologie orale, Lugano, 17 May 1995]. PMID- 7481692 TI - [Europe seen from Switzerland from the viewpoint of dentistry]. PMID- 7481695 TI - ["Women and men dentists must develop new ideas in order to win back the strength of conviction of earlier years"]. PMID- 7481694 TI - [Esthetics is the science of the beautiful (Duden). A report on the technical meeting of the Swiss Union for Pediatric Dentistry (SVK) of 17 May 1995 in Lugano]. PMID- 7481696 TI - [Fluorides in caries prevention. A report on the prophylaxis meeting of the School Dentistry Clinic Basel of 8 and 9 June 1995]. PMID- 7481697 TI - [The memory remains]. PMID- 7481699 TI - [The phagocytosis of polymorphonuclear neutrophilic granulocytes in progressive periodontitis]. AB - The aim of this paper was the evaluation of the phagocytic activity of neutrophils in blood and in gingival pocket fluid in patients suffering from rapidly progressive periodontitis (RPP) and postjuvenile periodontitis (PJP). Prior to periodontal treatment the authors evaluated the capacity to phagocytose latex particles of peripheral blood neutrophils from 21 patients with RPP, 51 with PJP and 59 healthy subjects (control group) as well as the phagocytic activity of neutrophils in pocket fluid from 21 patients with RPP, 14 with PJP and from 20 healthy subjects. This phagocytic activity was significantly lower in all examined groups in comparison with the control group. A similar evaluation executed 3 months after treatment revealed normal phagocytosis of blood neutrophils from patients with RPP. In patients receiving complementary pharmacotherapy (spiramycine combined with metronidazol), a better improvement of phagocytosis was noted, than that observed in patients treated only surgically. PMID- 7481702 TI - [Fissure caries. Their diagnosis and therapeutic principles]. PMID- 7481700 TI - [The microstructure and materials properties of provisional crown and bridge materials]. AB - The fracture resistance, surface hardness and microstructure of different manually and automatically mixed temporary crown and bridge resins and two light cured materials were examined in this study. The materials on butylmethacrylate (BMA) based showed low transverse strength compared to the materials on urethanmethacrylate (UDMA) and Bis-GMA base. Their values corresponded to the transverse strength of a manually mixed, chemically cured resin on PMMA base. The automatically mixed material on Bis-GMA base had lower strength values and surface hardness but fewer macroporosities compared to the manually mixed resins. The light cured material on UDMA base showed a transverse strength and surface hardness, which were higher than the values of the materials on Bis-GMA base. The strength values of the light cured material on PMMA base decreased within 7 days and were already after 24 h in the range of the materials on BMA base. PMID- 7481701 TI - [The reliability of apical x-ray pictures in the diagnosis of mandibular bone lesions. A review of the literature and in-vitro study]. AB - Using 6 macerated human anatomical preparations of mandibular jaw regions with either front teeth or premolars and molars, the radiological detectability of artificial periapical bone lesions was evaluated in relation to the size of the lesions and to the angle of x-ray projection. The artificial lesions were enlarged stepwise until distinct radiological visibility was attained. For each step of enlargement, x-ray pictures were taken with orthoradial as well as with at 25 degrees mesially and distally excentered projections. All x-ray pictures were evaluated by 6 dentists. The following results were obtained: The size of a periapical bone lesion, at which it is becoming radiologically detectable, varies between the different regions of the lower jaw. Isolated spongiosa lesions being larger than 3 mm in diameter are most often detectable at mandibular front teeth and premolars. Isolated spongiosa lesions at mandibular molars are generally non detectable. Atypical lesions, e.g. discontinuities of bony structures are particularly difficult to detect radiologically. There were no statistically significant differences in lesion detectability between x-rays of different angle projections. PMID- 7481698 TI - [The curing of composites under Cerec inlays]. AB - 96 Cerec CAD/CIM inlays (Vita Cerec Mk II, shades A2C and B3C) were seated into one reusable mod-cavity in a human molar. Light- (Brilliant Lux Incisal, Coltene) and dual- (Vita Duo Cement) curing composite resin respectively were used and the proximal areas were irradiated using a curing light (Coltolux II) with 4 different procedures: A) 210 s; B) 120 s (including 30 s Luciwedge, LW); C) 120 s (no LW) and D) 60 s. Vickers hardness (VHN) of the cured resin was measured at the pulpo-axial walls of the removed inlays. Polymerization-% of the resin was calculated relative to VHN of control samples, whose polymerization was scored as 100%. Two way Anova and Scheffe test were used for statistical comparison of the data. The results for the light-/dual-curing groups for shade A2C were: A) 97/89%, B) 89/90%, C) 91/88% D) 79/69%. Results for shade B3C were: A) 97/87%, B) 86/87%, C) 89/88% and D) 71/69%. No significant differences (p > 0.05) were found between light- and dual-curing. With procedures A, B and C light- and dual-curing resulted in adequate polymerization rates (PR > 85%) whereas D (PR < 80%) was significantly (p < 0.001) lower than A, B and C. Dual-curing resin had no advantages vs. light-curing with respect to polymerization rate when seating Cerec CAD/CIM inlays. The overall handling of the light-curing composite resin was judged to be easier than that of the dual cure material. PMID- 7481703 TI - [TENS in the face and neck areas. Transcutaneous electric nerve stimulation (TENS) in the therapy of chronic pain in the face and neck areas]. PMID- 7481704 TI - [The Federation Dentaire Internationale (FDI): a modern umbrella organization]. PMID- 7481706 TI - [The determination of the status of prosthodontics at Basel 1995]. PMID- 7481705 TI - [Changing pathways to oral health. A report on the symposium on the occasion of the retirement of Prof. Dr. Klaus H. Rateischak of 24 June 1995 in Basel]. PMID- 7481707 TI - [Esthetics in implantology. A report on the annual meeting of the Swiss Society for Implantology (SGI) of 17 May 1995 in the Congress-Center Lugano]. PMID- 7481708 TI - [Implants: the follow-up and treatment of peri-implant lesions. A report on the course "The follow-up of oral implants--treatment of peri-implant lesions" of 3 May 1995 in the Exhibits-Congress Center Basel]. PMID- 7481710 TI - [The 2nd National Congress for Dental Students. A report on the 2nd National Congress of the Association for Dental Students of Switzerland]. PMID- 7481711 TI - [Esthetics is the science of beauty (after Duden). A report on the professional meeting of the Association suisse de Pedodontie (ASP) of 17 May 1995 at Lugano]. PMID- 7481709 TI - [Ha-Ti user meeting. A report on the Ha-Ti-Dialog-Meeting 1994 in Freiburg im Breisgau]. PMID- 7481712 TI - [Fluoride in caries prevention. A report on the prophylaxis meeting of the Basel School Dentistry Clinic of 8 and 9 June 1995]. PMID- 7481713 TI - [The diagnostic specificity and sensitivity and the importance of disease prevalence]. AB - In this paper the terms "diagnostic specificity, sensitivity and efficiency" are explained using the example of feline immunodeficiency virus (FIV). Clinical conditions are discussed which require tests with high sensitivity and/or specificity. In addition it is shown that the prevalence of a disease is an important factor for the predictive value of a test result. For instance, tests with good specificity will lead to a high proportion of false negative values if the prevalence of the disease detected by the test is low. In order to improve the predictive value of a positive test result, the clinician must try to assign a patient to a population of individuals in which the prevalence of the suspected disease is elevated. This can be done by carefully evaluating the history and the clinical signs. PMID- 7481714 TI - [The control of hip dysplasia in Switzerland. A retrospect of the past 24 years]. AB - Examination of over 3700 dogs has proven that 42% of all purebred dogs in Switzerland are affected by CHD despite control programs for the last 30 years. Breeding with dysplastic dogs and lack of progeny control are responsible for this slow progress. PMID- 7481716 TI - [Ultrasonography of the adrenal gland in dogs]. AB - The technic for the ultrasonographic evaluation of the canine adrenal glands is described and illustrated. When specific preconditions are met, it is feasible to image both adrenal glands. Indications for adrenal ultrasonography are discussed. The normal ultrasonographic appearance and ultrasonographic features of adrenal disease are described. PMID- 7481715 TI - [Pectineus myectomy, tenotomy of the iliopsoas and neurectomy of the joint capsule as symptomatic therapy for coxarthrosis of the dog]. AB - Abduction and extension of the hip is reduced and painful in patients with coxarthrosis. Contracture of the pectineus and iliopsoas muscles restricts the abduction and extension of the hip, respectively. Pectineus myectomy and tenotomy of the iliopsoas muscle result in clinical improvements. This symptomatic therapy is relatively simple, with minimal invasion. Clinical results are satisfactory. PMID- 7481717 TI - [Enteral forced feeding using blind fixed stomach probes: two practical methods]. AB - Whereas fasting is well tolerated by the healthy organism, protein-energy malnutrition in the sick animal has a negative impact on organ functions, immunocompetence, wound healing, and reconvalescence. Sick animals, physically unable to eat or held off food for medical reasons for more than 5 days, consequently need some sort of nutritional support. Force feeding through a gastrostomy tube has several advantages compared to other forms of enteral nutrition. Gastrostomy tube feeding is relatively physiological, is quite comfortable for the animals and their owners, can replace oral feeding for weeks, and is associated with only rare complications. Gastrostomy tubes can be placed surgically, by use of a gastroscope, or blindly. Blind percutaneous gastrostomy tube placement is minimally invasive, fast, does not require any expensive equipment, and hence is well suited for practice. PMID- 7481718 TI - Suppurative pleuropneumonia and a pulmonary abscess in a ram: ultrasonographic and radiographic findings. AB - This report describes a two-year-old White Alpine ram with suppurative pleuropneumonia and a lung abscess. Prior to admission, the ram had been unsuccessfully treated with antibiotics and levamisole. Clinical examination revealed that the general behaviour and condition of the ram were severely disturbed. The rectal temperature and respiratory rate were increased. Auscultation of the lungs revealed increased vesicular sounds. Based on clinical findings, a tentative diagnosis of bronchopneumonia was made. To confirm the diagnosis, blood was taken for serological testing for Maedi-Visna, and endoscopic examination of the respiratory tract and ultrasonographic and radiographic examination of the thorax were performed. Cytologic and bacteriologic examination of tracheal secretions revealed large numbers of neutrophils and Actinomyces pyogenes organisms. A pocket of gas, surrounded by a capsule of soft tissue density, overlying the base of the heart, and a horizontal fluid line were observed on radiographs. Ultrasonographic examination revealed an effusion between the pleura and the lung on the left side of the thorax; an encapsulated abscess was seen on the right side of the thorax. Centesis and aspiration of this accumulation of fluid yielded foul-smelling pus. Post mortem examination confirmed the clinical, radiographic and ultrasonographic findings. The ram had severe chronic suppurative pleuropneumonia with abscess formation between the pleura and lung on the right side. PMID- 7481719 TI - The molecular logic of smell. AB - Mammals can recognize thousands of odors, some of which prompt powerful responses. Recent experiments illuminate how the nose and brain may perceive scents. PMID- 7481720 TI - Can environmental estrogens cause breast cancer? PMID- 7481721 TI - A pox on the pox. New vaccine raises hopes and doubts. PMID- 7481722 TI - Emerging viruses. AB - Hemorrhagic fever viruses are among the most dangerous biological agents known. New ones are discovered every year, and artificial as well as natural environmental changes are favoring their spread. PMID- 7481723 TI - Another episode in the continuing conflict. PMID- 7481724 TI - Wild death. PMID- 7481725 TI - MealMate: improving the nutritional status of elders using a milk-based nutritional supplement. AB - MealMate was a longitudinal research project conducted by the staff at the Geriatric Health Institute, a joint venture between Sioux Valley Hospital and the University of South Dakota School of Medicine. During Fall 1994, 64 older adults enrolled in this nutrition study at a Sioux Falls congregate dining site and drank a half-pint of whole milk combined with Carnation Instant Breakfast daily for one month. Pre-test and post-test data collected included anthropometric measures, specific blood tests from a venipuncture blood draw and use of standardized instruments to assess nutritional risk, depression, mental status and general demographic information. Results suggest that a longitudinal study with elders can be done effectively (attrition rate of less than 11%). While elders were very healthy at the onset, they nevertheless showed improved nutritional status over time. Laboratory tests showed significant increases in Vitamin D levels in adults over age 79. Using a more malnourished, home-bound sample of elders over age 79 and adding only whole milk to their diets are discussed as possible considerations for future research. PMID- 7481726 TI - A new class of antihypertensive: angiotensin II receptor antagonists. PMID- 7481727 TI - Validity criteria for exposure assessment methods. AB - The validity of any exposure assessment is an essential step in risk assessment. Following definitions of the various types of measurement validity, such as content, criterion and construct validity, and study validity, some shortcomings of actual exposure measurements are discussed. Both direct and indirect ways of measuring exposures to air, water and food pollutants are taken into consideration, with special attention being payed to indirect measurements using questionnaires and/or time-activity budgets. It is concluded that it is difficult to measure validity in an absolute way. Rather, a relative evaluation is possible comparing the validity of one result or study to that of another. PMID- 7481728 TI - Exposure assessment needs in studies of acute health effects. AB - It is known that pollutants in the outdoor and indoor environments are associated with acute adverse effects on health. The acute health effects produced include a wide range, from minor biological changes to causes of illness and exacerbation of disease, to death. Valid assessment of total exposures in various microenvironments are much more meaningful in terms of the various health responses. Short and long exposure times are both important, especially as related to the locations of such exposure and activities engaged in at the time. The quantitative character of exposure-response relationships will require more extensive research on obtaining appropriate exposure information, including monitoring and validating exposures and time activities, and on further modelling. More knowledge is also needed regarding the quantitative nature of the acute health effects, combined effects of different pollutants, and on the effects of various co-variates, effect-modifiers, and confounders. Efforts should also consist of promoting acute health effects studies that have appropriate monitoring and exposure assessment. PMID- 7481729 TI - Exposure assessment needs in studies of delayed health effects. AB - In studies of delayed effects of environmental exposures there is a need for accurate reconstruction of exposure, to the environmental factor under study, for a large number of individuals. There is also a need for similar assessments of exposure to possible confounding factors for the same population. In general, such reconstruction is not possible but it is often possible to make an estimate of exposures as they would have likely occurred. This requires selection of populations and circumstances which allow such estimates to be made. For each environmental exposure and for each disease outcome to be evaluated it is likely that the optimum exposure assessment methodology will be different. In each case these opportunities will have to be identified, tested and validated to the extent possible. Exposures will almost always have to be assessed retrospectively, if for no other reason than that prospective designs would not yield results for long periods of time after the initiation of the study. Examples are presented for the case of residential radon progeny exposure and associated excess lung cancer, where the delay is of the order of decades, and for the case of the possible teratogenic effects associated with living in the proximity of sites in which hazardous wastes are stored. In the latter case the delay is much shorter. PMID- 7481730 TI - Assessment of exposure to chemical contaminants in water and food. AB - The main questions on the validity of the studies and routine monitoring (e.g., in the scope of the WHO/UNEP GEMS programme) assessing exposure to chemicals through water and food (i.e., through digestion) relate to the following. (a) SAMPLING: variations due to sampling strategy and approach usually exceed analytical errors. (b) MEASUREMENT: quality assessment and quality control (QA/QC) procedures are essential. However, potential limitations of measurement errors resulting from implementation of these procedures should be kept in perspective with other errors and confounding factors. (c) METHODOLOGY: which depends on the variability of the intake patterns in the population, as well on the adequacy of the information on the consumption of food of a specified type as an indicator of the intake of a specific chemical. These factors should be considered in particular in studies using 'food basket' methods. Whenever possible, structure of the population in respect to the factors determining typical diet should be registered. (d) Form of reporting: limiting possibilities for re-analysis of the exposure estimates. In conclusion, the routinely collected data on exposure through food and water should be used with care in epidemiological studies. Assessment of the exposure conducted specially for the epidemiological study should consider possible changes of the diet with time. The data on confounders are crucial for exposure estimates. PMID- 7481731 TI - Assessment of dermal exposure to chemicals. AB - The methods for the dermal exposure assessment vary in their complexity and are in some sense complementary to each other. The most easy-to-use methods involve a pseudo-skin-approach, such as gloves and removal by washing. In some cases generic modelling appears to be possible. The experimental methods can indicate and even quantitate the presence of chemicals on the skin. This enables studies on the occurrence of local effects in relation to the exposure. When the interest is on systemic effects, the dermal exposure is only of interest if it represents the amount that is available for penetration through the skin. This may vary largely between compounds due to the large variation in dermal absorption. When this degree of absorption is not known, the alternate method may be biological monitoring, at least when it is based on a detailed pharmacokinetic knowledge of that compound. The most sophisticated method, applied to study occupational exposures, is formed by a combination of monitoring on clothing (pseudo-skin), hand washing (removal) and biological monitoring. In any case, the assessment of dermal exposure should be based on a sampling strategy that takes into account the distribution of the contamination on the body, the variation in time of the exposure, the duration of the exposure as well as the degree of skin protection afforded by clothing. PMID- 7481732 TI - Exposure assessment strategies in epidemiological studies of health effects of electric and magnetic fields. AB - In the growing epidemiological literature on potential health effects of electric and magnetic fields in homes, from appliances, and in the workplace, exposure assessment persists as the principal challenge. Associations of concern are residential exposure and childhood cancers, occupational exposure and adult leukemia and brain cancer, and video display terminal use and pregnancy outcome. The biologically appropriate measure of exposure is undefined, there are a multitude of exposure sources, and because the concern is with such rare diseases as leukemia and brain cancer, all studies have retrospective exposure ascertainment. Residential exposure assessment has been based on wire configuration codes of nearby electric power lines or on in-home measurements, both proxies for historical exposures of interest. Each has theoretical and logistical advantages and disadvantages. Appliance exposures are based on self report, focusing on those used for prolonged periods in close proximity to the user, such as electric blankets. Workplace exposures have been assessed by job title, more recently supplemented with measurements. Research is needed to: (1) develop and validate historical exposure models; (2) find ways to integrate exposure across diverse sources in the home and workplace; and (3) exploit laboratory findings to guide the selection of exposure metrics and relevant time windows. PMID- 7481733 TI - Methodology for assessment of exposure to environmental factors in application to epidemiological studies: assessment of exposure to natural ionizing radiation. AB - This paper reviews the recent progress of passive measurement methods of gamma radiation and radon. Relatively cheap and reliable dosimeters are now available. Results from national and world-wide surveys of natural radiation in bedrock and soils and indoor radon concentrations exist. These can be used to more precisely indicate areas with enhanced exposure to gamma radiation, indoor radon levels and radium and radon in household water. Besides direct measurements of the radiation intensity, additional information (e.g., history of residence, house ventilation habits) is required to reconstruct historical exposure levels for epidemiological investigations. The availability and correctness of this information may influence the validity of exposure estimates. PMID- 7481734 TI - Monitoring of exposure to air pollution. AB - This paper reviews recent advancements in methods of human exposure assessment to air pollutants. Instruments are becoming available for continuous recording of personal exposures, but their operating performance should be thoroughly assessed for accuracy, precision, interference, etc. Even after the method validation, it will be difficult and expensive to undertake continuous personal exposure monitoring in large study groups. The specification of exposures across a large population is probably best carried out using a combination of measurements and models. Detailed exposure measurements can be valuable in small-scale studies of exposure patterns, in pilot studies or to assess the validity of surrogate or proxy measures of exposure. PMID- 7481735 TI - Questionnaire based exposure assessment methods. AB - In environmental epidemiological studies, questionnaires may be the method of choice for assessing exposure because no other source of information is available, or because they provide the most efficient study design, allowing a larger study size and greater statistical power than would be possible with other more accurate measurement techniques. Problems may arise from a lack of content validity (the questionnaire does not cover all sources of exposure to the hazard of interest) or criterion validity (e.g., through inaccurate recall or misunderstanding of questions). Methods for validating questionnaires are described. The usefulness of validated 'standard' questionnaires is discussed, particularly in regard to comparison of exposure patterns in various populations studied separately. It is stressed that the validity of standard questionnaires should not be automatically assumed when they are applied to populations other than those in which they have been tested. PMID- 7481736 TI - Models of human exposure based on environmental monitoring. AB - The relevant exposure to environmental pollutants or relevant confounders can virtually never be measured directly in sufficient detail in a sufficient number of people included in an epidemiological study. Instead, surrogate indicators of exposure are used that are implicitly or explicitly linked by (conceptual) models to the 'relevant exposure'. Using specified indices (e.g., the coefficient of alienation and forecasting efficiency), the models of different forms can be compared and tested in special investigations. This can evaluate the criterion validity of the model through analysis of the agreement between the model estimates, based on measured surrogate indicators of the exposure, and the actual exposure. The perspective on the construct under study may further change the validity of the exposure indicator. Therefore, sub-studies evaluating the quality of exposure indicators should be considered as integral part of any study in environmental epidemiology, preferably as a pre-study or pilot study. PMID- 7481737 TI - Study design for exposure assessment in epidemiological studies. AB - We consider the implication, for study efficiency, of choice of method of exposure assessment in epidemiological studies, and in particular the optimal allocation of resources that should be devoted to improving the accuracy of exposure assessments. Useful for this purpose is a general result that the efficiency of a study based on approximate exposures relative to one based on exact exposures is equal to the square of the correlation between the true exposure and the approximate measurement in the study base (called the validity coefficient). This implies that to maximize study power, investment in increased precision is worthwhile up to the point at which proportional increase in total costs per subject exceed the proportional gain in the square of the validity coefficient. This result does not hold if exposure measurement error depends on disease status (is differential), or if important confounders are measured with error. 'Classical' exposure measurement error (uncorrelated with true exposure) or misclassification usually biases estimates of effect. Information from validity or reliability sub-studies can be used to correct this bias, but not substantially recover lost efficiency. There are several papers on the optimal allocation of resources to a validity sub-study. PMID- 7481738 TI - Methodology for assessment of exposure to environmental factors in application to epidemiological studies. WHO, regional office for Europe. AB - Assessment of exposure to environmental factors is a crucial step in each study investigating the impact of these factors on health, and is an essential component of the risk assessment process. The methodology of exposure assessment is developing rapidly, creating new opportunities for environmental epidemiology. At the same time, the practical requirements of population studies impose several restrictions on the feasibility of measurement methods, and determine certain requirements related to the method's validity. To evaluate existing exposure assessment methods in terms of their applicability to environmental epidemiology, the Consultation reviewed exposure assessment needs in studies of acute and delayed health effects, methods of assessing exposure through specific media or routes, and issues related to study design and exposure modelling. The recommendations refer to study design and conduct, methods of evaluation and publication, and to the policy issues and applications. PMID- 7481739 TI - Organochlorine residues in two seal species from western Iceland. AB - Harbour seals and grey seals from Faxafloi (Western Iceland) were analyzed for contamination with organochlorines (HCB, PCBs, p,p'-DDT,p,p'-DDE, chlordanes and the enantiomers of alpha-HCH). Although the values strongly varied, both harbour seals and grey seals on averaged showed comparable levels of PCBs, p,p'-DDT, p,p' DDE and alpha-HCH, as well as chlordanes (sigma CD). Differences were measured in the HCB content of the seal species. The HCB/alpha-HCH ratio in harbour seals was < 1 and in grey seals > 1. The results from seals on Iceland were compared with data derived from seal samples of the Antarctic, the Arctic (Spitzbergen) and the North Sea (Germany) in order to give an insight into the global distribution of pollution with chlorinated organics. PMID- 7481740 TI - Aluminum levels in human milk. AB - Aluminium levels in human milk in the winter period of 1992/93 ranged from 0.004 mg/l to 2.67 mg/l and the mean value was 0.38 mg/l. The nursing women were divided into several groups according to the results of a questionnaire, i.e. according to their social status (if they were refugees or not), number of deliveries, post partum day, the weight they put on during pregnancy, their age, and smoking habits. The mean levels of aluminium for each group were presented. The measurement of aluminium was done by graphite furnace atomic absorption spectrophotometry. PMID- 7481741 TI - Dietary mercury intake and human exposure in an Adriatic population. AB - A study was conducted to examine human exposure to mercury through dietary mercury intake in a population living in an industrially non-polluted area of the Adriatic Sea. The results have shown that approximately 20% of the subjects had a weekly dietary mercury intake above the provisional tolerable weekly intake (PTWI), primarily those consuming fish and other seafood > 6 times/week. The estimated seafood consumption corresponding to a mean intake of PTWI of 300 micrograms total mercury was 1559 g, and 1365 g for a PTWI of 200 micrograms methylmercury. However, the total mercury content in hair in individuals consuming total mercury above the PTWI was in the range of 1.3-12.9 micrograms/g, whereas the methylmercury content in hair in subjects consuming methylmercury above the PTWI was between 1.1-10.8 micrograms. Thus, the mercury content in hair did not reach the critical level at which toxic effects of mercury could be expected. The results, particularly those related to methylmercury exposure, did not differ significantly from data reported earlier from an industrially polluted area, thus indicating that the mercury content of fish and consequent human exposure to mercury reflects primarily the general ecological characteristics of the Adriatic, rather than the impact upon a specific local pollution. PMID- 7481742 TI - Microbial biosorption of copper and lead from aqueous systems. AB - Biosorption of metal ions from aqueous systems was evaluated using a culture of acidic soil isolates grown in a completely mixed, aerobic, semi-batch culture reactor. The laboratory scale system was used to test single and bimetallic solutions of copper and lead with sulfates, chlorides, or nitrates. To elucidate the key factors influencing biosorption and to characterize metal uptake by cellular and extra cellular components of the microbial system, a dialysis testing procedure was developed. A direct contact technique was used to determine the rate of metal sorption on cellular surfaces. The effectiveness of biosorption was influenced by pH, initial metal concentrations, and anionic composition. Respirometric tests were carried out to identify potential inhibitory effects of metal accumulation on microbial oxygen uptake rates. PMID- 7481744 TI - Environmental toxicology: hazards to the environment and man in the mediterranean region. Papers presented at a meeting of the Society of Ecotoxicology and Environmental Safety (SECOTOX). Rome, Italy, 26-29 September 1993. PMID- 7481745 TI - Biological monitoring of lead in workers exposed to tetraethyllead. AB - The excretion of inorganic, and total lead was investigated in the urine of workers who were exposed to tetraethyllead (TEL) at gasoline stations. Concentrations of total and inorganic lead after chelation-extraction were determined by flame atomic absorption spectrophotometry (FAAS) in combination with a slotted quartz tube. The limit of detection was 5.2 x 10(-3) micrograms Pb/ml; average total lead and inorganic lead concentrations in the urine of workers were 79.0 (range 22.6-158.9 micrograms Pb/g creatinine) and 37.3 micrograms Pb/g creatinine (range 5.1-121.0), respectively. In the control group, the mean of the urinary total lead and inorganic lead levels were 5.5 and 3.9 micrograms Pb/g creatinine, respectively. The difference between total and inorganic lead concentrations could be due to organic lead present in the urine. PMID- 7481743 TI - A possible path for mercury in biological systems: the oxidation of metallic mercury by molecular oxygen in aqueous solutions. AB - Metallic mercury has been assumed by several authors as not very reactive and, as a consequence, with little or no toxicity. The toxicity of this element is usually ascribed to alkyl-mercury ions considered to be formed by some microorganisms. In this work, we describe experiments that clearly show that metallic mercury can be easily oxidized by molecular oxygen in aqueous solution in the presence of species such as chloride, which complex Hg(II). The experiments were carried out using metallic mercury in NaCl aqueous solution under 'open air' (temperature and agitation rate maintained constant) and under more controlled conditions (CO2 rate bubbling, i.e. pH = 4.2; air rate bubbling, i.e., O2 constant concentration, temperature, agitation rate). The reactions were monitored spectrophotometricaly at 230 nm (HgCl2-(4)). Significative values of the concentration of Hg(II) in the form of HgCl2-(4) were soon attained in those solutions. For example, in 'open air' conditions, at 25 degrees C and [NaCl] = 30 g/l (0.51 mol/dm3), the maximal concentration of 13 ppm (6.44 x 10(-5) mol/dm3) of Hg(II) in the form of HgCl2-(4) was reached in 120 min; for [NaCl] = 5 g/l at 25 degrees C, (0.085 mol/dm3) the maximal concentration of 0.3 ppm (1.53 x 10(-6) mol/dm3) of Hg(II) in the form of HgCl2-(4) was reached in 10 min. The rate constants, kobs, of the oxidation of the metallic mercury under the studied conditions are pseudo zero-order at 25 degrees C, and under more controlled conditions have ranged from 1.0 x 10(-7) mol/min ([NaCl] = 5 g/l identical to 0.085 mol/dm3) to 20.0 x 10(-7) mol/min ([NaCl] = 300 g/l identical to 5.12 mol/dm3). The rate constant increases with temperature, up to 25 degrees C, from where kobs remains constant up to 40 degrees C. From the analysis of the experimental results it was possible to propose a mechanism of oxidation of metallic mercury by O2 in aqueous solution containing NaCl. This oxidation is proposed as a possible route for the introduction of mercury into biological systems. PMID- 7481746 TI - Biochemical biomarkers in ecotoxicology--some recent developments. AB - Biochemical biomarkers measure the exposure of organisms to environmental chemicals. They can also provide measures of toxic effect, e.g. where they are based on molecular mechanisms which underly toxicity. Ideally, biomarkers should be sensitive, specific, simple to use and suitable for the assay of material obtained by non-destructive sampling procedures (e.g. of blood). Recently, there has been encouraging progress in the development of several different types of biomarker assays: (1) The measurement of inhibition of serum 'B' esterases to monitor exposure of birds to organophosphorus insecticides. (2) The measurement of DNA damage caused by aromatic hydrocarbons. DNA adduct formation has been studied using the 32P-postlabelling technique. Several other techniques are currently under investigation. (3) The measurement of disturbances to the transthyretin-retinol binding protein complex caused by a metabolite of 3,4,3',4',tetrachlorobiphenyl. (4) The measurement of precursors of clotting proteins in blood following the inhibition of the Vitamin K cycle by anticoagulant rodenticides. Of these examples, the first is only a biomarker of exposure but the remaining three examples are, in principle, biomarkers of toxic effect since they all represent measures of molecular mechanisms which underly toxicity. Biochemical biomarkers have considerable potential for measuring effects of chemicals under field conditions--especially where carefully selected combinations of them are used. PMID- 7481747 TI - The study of interactive effects of pollutants: a biomarker approach. AB - Biochemical biomarkers, such as inhibition of serum butyryl cholinesterase (BuChE) and brain acetyl cholinesterase (AChE), have been useful in studies of interactive effects of pesticides in birds. Examples of interactions due to increased activation or decreased detoxication are reviewed. Studies have shown that hybrid red-legged partridges (Alectoris rufa cross) pretreated with the inducing ergosterol biosynthesis inhibiting (EBI) fungicide, prochloraz, were more sensitive to the toxic effects of the organophosphorous (OP) insecticide, malathion, than controls. A dose of 90 mg/kg prochloraz produced greater inhibition at 1, 4 and 24 h following oral administration of 50 mg/kg malathion, compared to corn oil controls. Pigeons (Columba livia) given 180 or 90 mg/kg prochloraz showed greater inhibition of BuChE activity following malathion administration than did control birds. Starlings (Stumus vulgaris), however, appeared not to be induced by 180 or 300 mg/kg prochloraz, and no difference in BuChE activity following dosing with malathion was apparent in comparison with controls. Other EBIs and OP combinations have been investigated in the partridge. Birds pretreated with prochloraz showed a trend towards greater inhibition of serum BuChE activity at most time points following dosing with the OPs dimethoate and chlorpyriphos. Birds pretreated with the EBI penconazole showed significantly greater inhibition of serum BuChE activity at 1, 4 and 24 h after malathion administration than did controls. The mechanism of increased activation of malathion due to induction of cytochrome P-450 by prochloraz is reviewed. In the case of interactions due to inhibition of detoxication, inhibition of brain AChE activity was a useful biochemical biomarker.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7481748 TI - Enzymological differences of AChE and diazinon hepatic metabolism: correlation of in vitro data with the selective toxicity of diazinon to fish species. AB - The in vitro hepatic metabolism of diazinon, as well as the sensitivity of the brain acetylcholine esterase, to diazoxon inhibitory action have been studied in order to explain the different toxicity of diazinon to Oncorhynchus mykiss (rainbow trout), Poecilia reticulata (guppy), Brachydanio rerio (zebra fish) and Cyprinus carpio (carp). In spite of a very sensitive acetylcholine esterase the carp is very resistant to diazinon toxicity because of its very low rate of bioactivation and relatively high activity of detoxicating enzymes. The trout is very sensitive towards diazinon in spite of its low activity of bioactivation, because of its lack of detoxicating enzymes and a very sensitive acetylcholine esterase. Diazinon is very toxic for the guppy, because this fish combines a relatively sensitive acetylcholine esterase with a high rate of bioactivation. The zebra fish has the most insensitive acetylcholine esterase, associated with a limited activation rate, thus resulting a rather resistant species. The results obtained indicate that diazinon toxicity differences among the fish species studied can largely be explained in relation to metabolic balances in the liver and with the features of the target enzyme. PMID- 7481749 TI - Immobilised yeast cells biosensor for total toxicity testing. AB - An immobilised yeast cell biosensor has been developed for the total toxicity testing of a sample that may contain a number of different pollutant species; the biosensor uses an amperometric gas diffusion oxygen as indicator electrode. The method is based on the perturbation of the respiratory activity of a yeast, Saccharomyces cerevisiae, immobilised on an agar gel containing the culture medium (i.e., 'agarised medium'), by the toxic test substance. Glucose is used as substrate while the toxic substances tested consist of several metallic ions, phenol and cationic or anionic surfactants. PMID- 7481750 TI - Drinking water quality and monitoring in north Africa: the Moroccan experience. AB - In Morocco, the water quality standards have been derived from World Health Organization guidelines for all that concerns bacteriological and toxicological parameters; for the rest of the parameters, several other sources have been used especially EEC, US and Canadian standards. Local considerations have dictated, however, the value of the maximum acceptable level adopted for the total dissolved solids (2000 mg/l), instead of the 1500 mg/l generally considered to be the maximum acceptable level. The National Office of Potable Water (ONEP) developed quite an extensive network for water quality monitoring, comprised of a central laboratory conducting all sorts of analytical determinations ranging from parasitological, planktonic and bacteriological examinations, to the determination of trace metals, pesticides, global radioactivity and toxicity tests. The work of the central laboratory is seconded by 34 small branch laboratories spread all over the country. The preparation of reference analytical methodologies has been a subject on the agenda of the next phase of the work of the commission in charge of the preparation of standards dealing with drinking water. A first approach of the Moroccan water quality standards implementation have been made by ONEP. The evaluation of the costs necessary to be in conformity with these standards and the dispensation requests to be presented, where appropriate, to health authorities are on the way. PMID- 7481751 TI - The evolution of groundwater quality in France: perspectives for enduring use for human consumption. AB - France is rich in groundwater. It has many aquifers with renewable resources which are estimated at 100 billion m3/year, of which 3.5 billion m3, or 60% of the water used in France for human consumption, are withdrawn each year. This practice is justified by the often naturally pure quality of the water and by the regularity of the resource. Nevertheless, free aquifers, in spite of natural physical and geochemical barriers, are not sufficiently protected from anthropic surface contamination and when pollutants reach them, the consequences are never negligible, be it from a sanitary, economic or natural heritage point of view. The most extensive pollution is nitrate contamination. Nitrate concentrations have been increasing constantly over the last 30 years and in some regions have gone over the critical threshold of 50 mg/l which is the European standard, and concentrations of 100 mg/l have been measured in some places. The gravity of other types of pollution--mining, industrial or domestic--is determined by the mineral or organic products involved. The cumulative effect of all of this pollution is serious from various standpoints: from an economic and environmental point of view because aquifers contribute to the maintenance of aquatic life, and for our natural heritage because of the long-term degradation of vast aquifers in some very urban and industrial regions. We might, therefore, question the possible long-term use of aquifers as a source of drinking water. PMID- 7481752 TI - Use of computer models to assess exposure to agricultural chemicals via drinking water. AB - Surveys of drinking water quality throughout the agricultural regions of the world have revealed the tendency of certain crop protection chemicals to enter water supplies. Fortunately, the trace concentrations that have been detected are generally well below the levels thought to have any negative impact on human health or the environment. However, the public expects drinking water to be pristine and seems willing to bear the costs involved in further regulating agricultural chemical use in such a way so as to eliminate the potential for such materials to occur at any detectable level. Of all the tools available to assess exposure to agricultural chemicals via drinking water, computer models are one of the most cost-effective. Although not sufficiently predictive to be used in the absence of any field data, such computer programs can be used with some degree of certainty to perform quantitative extrapolations and thereby quantify regional exposure from field-scale monitoring information. Specific models and modeling techniques will be discussed for performing such exposure analyses. Improvements in computer technology have recently made it practical to use Monte Carlo and other probabilistic techniques as a routine tool for estimating human exposure. Such methods make it possible, at least in principle, to prepare exposure estimates with known confidence intervals and sufficient statistical validity to be used in the regulatory management of agricultural chemicals. PMID- 7481753 TI - Investigating groundwater pollution from different sources with combined biological and chemical methods. AB - This paper reviews groundwater pollution caused by the disposal of untreated effluents of a dye factory located 20 km to the west of Nicosia (Cyprus). The task of the work was to investigate the nature of the pollution and differentiate it from other possible pollution sources in the area. It focused on toxicity testing and biofractionation in order to address the most toxic pollutants and, on the evaluation of GC/FID profiles for investigating the connection between groundwater pollution and the effluents. This connection was successful due to a multiple comparison amongst the GC/FID profiles resulting from the polluted groundwater, the water from the reference areas and the dye effluents. PMID- 7481754 TI - Trihalomethanes in drinking water and cancer: risk assessment and integrated evaluation of available data, in animals and humans. AB - In our study, we attempted to jointly consider THM concentration data collected from drinking waters and carcinogenic risk assessment derived from mathematical models commonly used in this field (multi-stage models for laboratory animal experimentation data, and 'unit risk' derived from the relative risk in the case of epidemiological data). In order to estimate the risks related to joint exposure to different THMs, in this study the risk additivity hypothesis is taken into account. Based on animal data for the various tumors, carcinogenic risk estimates for different THM combinations vary from 2.7 x 10(-7) to 4.6 x 10(-6) per micrograms/l in relation to different carcinogenic substances published in the literature or specifically calculated in this study. The carcinogenic risk parameters derived from experimental studies and from epidemiological data were substantially consistent. Our study uses also as an example some data on concentration levels of THMs for drinking water supplies in Sardinia. The area mean THM concentration values for each supply varied, for ground waters, from 8.1 to 13.6 micrograms/l and, for surface waters, from 52.8 to 168 micrograms/l. For the 1976-1989 period, bladder cancer standardized mortality rates in the water distribution system areas where the THMs were measured indicate values similar, but generally lower, than the national ones, except in the province of Cagliari where the values were not significantly different. The risk estimates derived from animal studies are of the same order of magnitude as the epidemiological data in literature. PMID- 7481755 TI - Women's longevity. PMID- 7481756 TI - MHC class I gene expression. PMID- 7481757 TI - Antibiotics that resist resistance. PMID- 7481758 TI - Patents. New biotech law shores up U.S. firms. PMID- 7481759 TI - New clock gene cloned. PMID- 7481760 TI - Muscular dystrophies: diseases of the dystrophin-glycoprotein complex. PMID- 7481761 TI - Cracking the neuronal code. PMID- 7481762 TI - Development and plasticity of cortical processing architectures. AB - One of the basic functions of the cerebral cortex is the analysis and representation of relations among the components of sensory and motor patterns. It is proposed that the cortex applies two complementary strategies to cope with the combinatorial problem posed by the astronomical number of possible relations: (i) the analysis and representation of frequently occurring, behaviorally relevant relations by groups of cells with fixed but broadly tuned response properties; and (ii) the dynamic association of these cells into functionally coherent assemblies. Feedforward connections and reciprocal associative connections, respectively, are thought to underlie these two operations. The architectures of both types of connections are susceptible to experience dependent modifications during development, but they become fixed in the adult. As development proceeds, feedforward connections also appear to lose much of their functional plasticity, whereas the synapses of the associative connections retain a high susceptibility to use-dependent modifications. The reduced plasticity of feedforward connections is probably responsible for the invariance of cognitive categories acquired early in development. The persistent adaptivity of reciprocal connections is a likely substrate for the ability to generate representations for new perceptual objects and motor patterns throughout life. PMID- 7481763 TI - The brain's visual world: representation of visual targets in cerebral cortex. AB - Microelectrode recordings from behaving monkeys have shown that neuronal responses in the visual cerebral cortex can depend greatly on which aspect of the scene is the target of the animal's attention. Accumulating evidence suggests that while the early stages of the visual pathway provide a faithful representation of the retinal image, later stages of processing in the visual cortex hold representations that emphasize the viewer's current interest. By filtering out irrelevant signals and adding information about objects whose presence is remembered or inferred, the cortex creates an edited representation of the visual world that is dynamically modified to suit the immediate goals of the viewer. PMID- 7481764 TI - Functional brain imaging studies of cortical mechanisms for memory. AB - Recent functional brain imaging studies in humans indicate that learning and memory involve many of the same regions of the cortex that process sensory information and control motor output. The forms of perceptual and motor learning that can occur without conscious recollection are mediated in part by contractions and expansions of representations in the sensory and motor cortex. The same regions are also engaged during the conscious storage and retrieval of facts and events, but these types of memory also bring into play structures involved in the active maintenance of memories "on line" and in the establishment of associative links between the information stored in different sensory areas. Although the picture of memory that is emerging from functional imaging studies is consistent with current physiological accounts, there are puzzles and surprises that will be solved only through a combination of human and animal studies. PMID- 7481765 TI - Aberrant subcellular localization of BRCA1 in breast cancer. AB - The BRCA1 gene product was identified as a 220-kilodalton nuclear phosphoprotein in normal cells, including breast ductal epithelial cells, and in 18 of 20 tumor cell lines derived from tissues other than breast and ovary. In 16 of 17 breast and ovarian cancer lines and 17 of 17 samples of cells obtained from malignant effusions, however, BRCA1 localized mainly in cytoplasm. Absence of BRCA1 or aberrant subcellular location was also observed to a variable extent in histological sections of many breast cancer biopsies. These findings suggest that BRCA1 abnormalities may be involved in the pathogenesis of many breast cancers, sporadic as well as familial. PMID- 7481766 TI - Orphanin FQ: a neuropeptide that activates an opioidlike G protein-coupled receptor. AB - A heptadecapeptide was identified and purified from porcine brain tissue as a ligand for an orphan heterotrimeric GTP-binding protein (G protein)-coupled receptor (LC132) that is similar in sequence to opioid receptors. This peptide, orphanin FQ, has a primary structure reminiscent of that of opioid peptides. Nanomolar concentrations of orphanin FQ inhibited forskolin-stimulated adenylyl cyclase activity in cells transfected with LC132. This inhibitory activity was not affected by the addition of opioid ligands, nor did the peptide activate opioid receptors. Orphanin FQ bound to its receptor in a saturable manner and with high affinity. When injected intracerebroventricularly into mice, orphanin FQ caused a decrease in locomotor activity but did not induce analgesia in the hot-plate test. However, the peptide produced hyperalgesia in the tail-flick assay. Thus, orphanin FQ may act as a transmitter in the brain by modulating nociceptive and locomotor behavior. PMID- 7481767 TI - Defects in B lymphocyte maturation and T lymphocyte activation in mice lacking Jak3. AB - Biochemical studies of signaling mediated by many cytokine and growth factor receptors have implicated members of the Jak family of tyrosine kinases in these pathways. Specifically, Jak3 has been shown to be associated with the interleukin 2 (IL-2) receptor gamma chain, a component of the receptors for IL-2, IL-4, IL-7, IL-9, and IL-15. Mice lacking Jak3 showed a severe block in B cell development at the pre-B stage in the bone marrow. In contrast, although the thymuses of these mice were small, T cell maturation progressed relatively normally. In response to mitogenic signals, peripheral T cells in Jak3-deficient mice did not proliferate and secreted small amounts of IL-2. These data demonstrate that Jak3 is critical for the progression of B cell development in the bone marrow and for the functional competence of mature T cells. PMID- 7481768 TI - Mutation of Jak3 in a patient with SCID: essential role of Jak3 in lymphoid development. AB - Males with X-linked severe combined immunodeficiency (XSCID) have defects in the common cytokine receptor gamma chain (gamma c) gene that encodes a shared, essential component of the receptors of interleukin-2 (IL-2), IL-4, IL-7, IL-9, and IL-15. The Janus family tyrosine kinase Jak3 is the only signaling molecule known to be associated with gamma c, so it was hypothesized that defects in Jak3 might cause an XSCID-like phenotype. A girl with immunological features indistinguishable from those of XSCID was therefore selected for analysis. An Epstein-Barr virus (EBV)-transformed cell line derived from her lymphocytes had normal gamma c expression but lacked Jak3 protein and had greatly diminished Jak3 messenger RNA. Sequencing revealed a different mutation on each allele: a single nucleotide insertion resulting in a frame shift and premature termination in the Jak3 JH4 domain and a nonsense mutation in the Jak3 JH2 domain. The lack of Jak3 expression correlated with impaired B cell signaling, as demonstrated by the inability of IL-4 to activate Stat6 in the EBV-transformed cell line from the patient. These observations indicate that the functions of gamma c are dependent on Jak3 and that Jak3 is essential for lymphoid development and signaling. PMID- 7481769 TI - Defective lymphoid development in mice lacking Jak3. AB - The Janus tyrosine kinases (Jaks) play a central role in signaling through cytokine receptors. Although Jak1, Jak2, and Tyk2 are widely expressed, Jak3 is predominantly expressed in hematopoietic cells and is known to associate only with the common gamma (gamma c) chain of the interleukin (IL)-2, IL-4, IL-7, IL 9, and IL-15 receptors. Homozygous mutant mice in which the Jak3 gene had been disrupted were generated by gene targeting. Jak3-deficient mice had profound reductions in thymocytes and severe B cell and T cell lymphopenia similar to severe combined immunodeficiency disease (SCID), and the residual T cells and B cells were functionally deficient. Thus, Jak3 plays a critical role in gamma c signaling and lymphoid development. PMID- 7481770 TI - Superior parietal cortex activation during spatial attention shifts and visual feature conjunction. AB - Positron emission tomography was used to measure changes in the regional cerebral blood flow of normal people while they searched visual displays for targets defined by color, by motion, or by a conjunction of color and motion. A region in the superior parietal cortex was activated only during the conjunction task, at a location that had previously been shown to be engaged by successive shifts of spatial attention. Correspondingly, the time needed to detect a conjunction target increased with the number of items in the display, which is consistent with the use of a mechanism that successively analyzes each item in the visual field. PMID- 7481771 TI - Positional cloning and sequence analysis of the Drosophila clock gene, timeless. AB - The Drosophila genes timeless (tim) and period (per) interact, and both are required for production of circadian rhythms. Here the positional cloning and sequencing of tim are reported. The tim gene encodes a previously uncharacterized protein of 1389 amino acids, and possibly another protein of 1122 amino acids. The arrhythmic mutation tim01 is a 64-base pair deletion that truncates TIM to 749 amino acids. Absence of sequence similarity to the PER dimerization motif (PAS) indicates that direct interaction between PER and TIM would require a heterotypic protein association. PMID- 7481772 TI - Rhythmic expression of timeless: a basis for promoting circadian cycles in period gene autoregulation. AB - The clock gene timeless (tim) is required for circadian rhythmicity in Drosophila. The accumulation of tim RNA followed a circadian rhythm, and the phase and period of the tim RNA rhythm were indistinguishable from those that have been reported for per. The tim RNA oscillations were found to be dependent on the presence of PER and TIM proteins, which demonstrates feedback control of tim by a mechanism previously shown to regulate per expression. The cyclic expression of tim appears to dictate the timing of PER protein accumulation and nuclear localization, suggesting that tim promotes circadian rhythms of per and tim transcription by restricting per RNA and PER protein accumulation to separate times of day. PMID- 7481773 TI - Isolation of timeless by PER protein interaction: defective interaction between timeless protein and long-period mutant PERL. AB - The period (per) gene likely encodes a component of the Drosophila circadian clock. Circadian oscillations in the abundance of per messenger RNA and per protein (PER) are thought to arise from negative feedback control of per gene transcription by PER. A recently identified second clock locus, timeless (tim), apparently regulates entry of PER into the nucleus. Reported here are the cloning of complementary DNAs derived from the tim gene in a two-hybrid screen for PER interacting proteins and the demonstration of a physical interaction between the tim protein (TIM) and PER in vitro. A restricted segment of TIM binds directly to a part of the PER dimerization domain PAS. PERL, a mutation that causes a temperature-sensitive lengthening of circadian period and a temperature-sensitive delay in PER nuclear entry, exhibits a temperature-sensitive defect in binding to TIM. These results suggest that the interaction between TIM and PER determines the timing of PER nuclear entry and therefore the duration of part of the circadian cycle. PMID- 7481774 TI - Participation of the human beta-globin locus control region in initiation of DNA replication. AB - The human beta-globin locus control region (LCR) controls the transcription, chromatin structure, and replication timing of the entire locus. DNA replication was found to initiate in a transcription-independent manner within a region located 50 kilobases downstream of the LCR in human, mouse, and chicken cells containing the entire human beta-globin locus. However, DNA replication did not initiate within a deletion mutant locus lacking the sequences that encompass the LCR. This mutant locus replicated in the 3' to 5' direction. Thus, interactions between distantly separated sequences can be required for replication initiation, and factors mediating this interaction appear to be conserved in evolution. PMID- 7481776 TI - Does the p53 up-regulated Gadd45 protein have a role in excision repair? PMID- 7481775 TI - Mutations in the dystrophin-associated protein gamma-sarcoglycan in chromosome 13 muscular dystrophy. AB - Severe childhood autosomal recessive muscular dystrophy (SCARMD) is a progressive muscle-wasting disorder common in North Africa that segregates with microsatellite markers at chromosome 13q12. Here, it is shown that a mutation in the gene encoding the 35-kilodalton dystrophin-associated glycoprotein, gamma sarcoglycan, is likely to be the primary genetic defect in this disorder. The human gamma-sarcoglycan gene was mapped to chromosome 13q12, and deletions that alter its reading frame were identified in three families and one of four sporadic cases of SCARMD. These mutations not only affect gamma-sarcoglycan but also disrupt the integrity of the entire sarcoglycan complex. PMID- 7481778 TI - Great transitions. PMID- 7481777 TI - Does the p53 up-regulated Gadd45 protein have a role in excision repair? PMID- 7481780 TI - Earth's carrying capacity. PMID- 7481779 TI - Earth's carrying capacity. PMID- 7481781 TI - AZT trial in Thailand. PMID- 7481782 TI - Biopolymer sequence availability. PMID- 7481783 TI - Dioxin research in Vietnam. PMID- 7481784 TI - Thailand weighs AIDS vaccine tests. PMID- 7481786 TI - Publicity fears cancel gene talk. PMID- 7481785 TI - IL-12 deaths: explanation and a puzzle. PMID- 7481787 TI - Tamoxifen's trials and tribulations. PMID- 7481788 TI - Monoclonal antibodies at age 20: promise at last? PMID- 7481789 TI - Sifting mitosis, cell fate in fly eyes. PMID- 7481791 TI - Asian anthropoids strike back. PMID- 7481790 TI - New clues found to how some people live with HIV. PMID- 7481792 TI - All about AIDS. PMID- 7481793 TI - Urgently needed: policies on access to data by erstwhile collaborators. PMID- 7481794 TI - Pharmacia Biotech & Science prize. Grand prize winner. Life and death decisions: ced-9 and programmed cell death in Caenorhabditis elegans. PMID- 7481795 TI - The Yin and Yang of T cell costimulation. PMID- 7481796 TI - Trimeric G proteins: surprise witness tells a tale. PMID- 7481797 TI - Remembering x-rays. PMID- 7481798 TI - Protein design: a hierarchic approach. AB - The de novo design of peptides and proteins has recently emerged as an approach for investigating protein structure and function. Designed, helical peptides provide model systems for dissecting and quantifying the multiple interactions that stabilize secondary structure formation. De novo design is also useful for exploring the features that specify the stoichiometry and stability of alpha helical coiled coils and for defining the requirements for folding into structures that resemble native, functional proteins. The design process often occurs in a series of discrete steps. Such steps reflect the hierarchy of forces required for stabilizing tertiary structures, beginning with hydrophobic forces and adding more specific interactions as required to achieve a unique, functional protein. PMID- 7481799 TI - Tertiary and quaternary structural changes in Gi alpha 1 induced by GTP hydrolysis. AB - Crystallographic analysis of 2.2 angstrom resolution shows that guanosine triphosphate (GTP) hydrolysis triggers conformational changes in the heterotrimeric G-protein alpha subunit, Gi alpha 1. The switch II and switch III segments become disordered, and linker II connecting the Ras and alpha helical domains moves, thus altering the structures of potential effector and beta gamma binding regions. Contacts between the alpha-helical and Ras domains are weakened, possibly facilitating the release of guanosine diphosphate (GDP). The amino and carboxyl termini, which contain receptor and beta gamma binding determinants, are disordered in the complex with GTP, but are organized into a compact microdomain on GDP hydrolysis. The amino terminus also forms extensive quaternary contacts with neighboring alpha subunits in the lattice, suggesting that multimers of alpha subunits or heterotrimers may play a role in signal transduction. PMID- 7481800 TI - Crystal structure of the biphenyl-cleaving extradiol dioxygenase from a PCB degrading pseudomonad. AB - Polychlorinated biphenyls (PCBs) typify a class of stable aromatic pollutants that are targeted by bioremediation strategies. In the aerobic degradation of biphenyl by bacteria, the key step of ring cleavage is catalyzed by an Fe(II) dependent extradiol dioxygenase. The crystal structure of 2,3-dihydroxybiphenyl 1,2-dioxygenase from a PCB-degrading strain of Pseudomonas cepacia has been determined at 1.9 angstrom resolution. The monomer comprises amino- and carboxyl terminal domains. Structural homology between and within the domains reveals evolutionary relationships within the extradiol dioxygenase family. The iron atom has five ligands in square pyramidal geometry: one glutamate and two histidine side chains, and two water molecules. PMID- 7481801 TI - Guidelines for protein design: the energetics of beta sheet side chain interactions. AB - To determine the interaction energy between cross-strand pairs of side chains on an antiparallel beta sheet, pairwise amino acid substitutions were made on the solvent-exposed face of the B1 domain of streptococcal protein G. The measured interaction energies were substantial (1.8 kilocalories per mole) and comparable to the magnitude of the beta sheet propensities. The experimental results paralleled the statistical frequency with which the residue pairs are found in beta sheets of known structure. PMID- 7481802 TI - Uncoupling cell fate determination from patterned cell division in the Drosophila eye. AB - Cell proliferation and cell fate specification are under strict spatiotemporal control in the developing Drosophila eye. Cells excluded from five-cell preclusters synchronously enter a single additional cell cycle, the second mitotic wave, after which the remaining cells are sequentially recruited. When the second mitotic wave was blocked with the human cyclin-dependent kinase inhibitor p21CIP1/WAF1, each cell type was still specified. Hence, cell fate determination is regulated independently of the division pattern of precursor cells. However, the second mitotic wave is needed to generate appropriate numbers of each cell type. Moreover, p21 can arrest precursor cell proliferation and allow appropriate fate choice in vivo. PMID- 7481803 TI - Lymphoproliferative disorders with early lethality in mice deficient in Ctla-4. AB - The role of the cell-surface molecule CTLA-4 in the regulation of T cell activation has been controversial. Here, lymph nodes and spleens of CTLA-4 deficient mice accumulated T cell blasts with up-regulated activation markers. These blast cells also infiltrated liver, heart, lung, and pancreas tissue, and amounts of serum immunoglobulin were elevated. The mice invariably became moribund by 3 to 4 weeks of age. Although CTLA-4-deficient T cells proliferated spontaneously and strongly when stimulated through the T cell receptor, they were sensitive to cell death induced by cross-linking of the Fas receptor and by gamma irradiation. Thus, CTLA-4 acts as a negative regulator of T cell activation and is vital for the control of lymphocyte homeostasis. PMID- 7481804 TI - Genomic structure of an attenuated quasi species of HIV-1 from a blood transfusion donor and recipients. AB - A blood donor infected with human immunodeficiency virus-type 1 (HIV-1) and a cohort of six blood or blood product recipients infected from this donor remain free of HIV-1-related disease with stable and normal CD4 lymphocyte counts 10 to 14 years after infection. HIV-1 sequences from either virus isolates or patient peripheral blood mononuclear cells had similar deletions in the nef gene and in the region of overlap of nef and the U3 region of the long terminal repeat (LTR). Full-length sequencing of one isolate genome and amplification of selected HIV-1 genome regions from other cohort members revealed no other abnormalities of obvious functional significance. These data show that survival after HIV infection can be determined by the HIV genome and support the importance of nef or the U3 region of the LTR in determining the pathogenicity of HIV-1. PMID- 7481805 TI - Converting Escherichia coli RNA polymerase into an enhancer-responsive enzyme: role of an NH2-terminal leucine patch in sigma 54. AB - The protein sigma 54 associates with Escherichia coli core RNA polymerase to form a holoenzyme that binds promoters but is inactive in the absence of enhancer activation. Here, mutants of sigma 54 enabled polymerases to transcribe without enhancer protein and adenosine triphosphate. The mutations are in leucines within the NH2-terminal glutamine-rich domain of sigma 54. Multiple leucine substitutions mimicked the effect of enhancer protein, which suggests that the enhancer protein functions to disrupt a leucine patch. The results indicate that sigma 54 acts both as an inhibitor of polymerase activity and as a receptor that interacts with enhancer protein to overcome this inhibition, and that these two activities jointly confer enhancer responsiveness. PMID- 7481806 TI - Transition in specification of embryonic metazoan DNA replication origins. AB - In early Xenopus embryos, in which ribosomal RNA genes (rDNA) are not transcribed, rDNA replication initiates and terminates at 9- to 12-kilobase pair intervals, with no detectable dependence on specific DNA sequences. Resumption of ribosomal RNA (rRNA) synthesis at late blastula and early gastrula is accompanied by a specific repression of replication initiation within transcription units; the frequency of initiation within intergenic spacers remains as high as in early blastula. These results demonstrate that for rRNA genes, circumscribed zones of replication initiation emerge in intergenic DNA during the time in metazoan development when the chromatin is remodeled to allow gene transcription. PMID- 7481809 TI - Genetic discrimination. PMID- 7481808 TI - Genetic discrimination. PMID- 7481807 TI - A left-handed parallel beta helix in the structure of UDP-N-acetylglucosamine acyltransferase. AB - UDP-N-acetylglucosamine 3-O-acyltransferase (LpxA) catalyzes the transfer of (R) 3-hydroxymyristic acid from its acyl carrier protein thioester to UDP-N acetylglucosamine. LpxA is the first enzyme in the lipid A biosynthetic pathway and is a target for the design of antibiotics. The x-ray crystal structure of LpxA has been determined to 2.6 angstrom resolution and reveals a domain motif composed of parallel beta strands, termed a left-handed parallel beta helix (L beta H). This unusual fold displays repeated violations of the protein folding constraint requiring right-handed crossover connections between strands of parallel beta sheets and may be present in other enzymes that share amino acid sequence homology to the repeated hexapeptide motif of LpxA. PMID- 7481810 TI - Genetic discrimination. PMID- 7481811 TI - E-mail privacy. PMID- 7481812 TI - Intrachain calcium binding. PMID- 7481813 TI - Varmus puts his stamp on NIH. PMID- 7481814 TI - Limb development. Gene ties arthropods together. PMID- 7481815 TI - Sexing fossils: a boy named Lucy? PMID- 7481816 TI - Dogfight erupts over animal studies in the Serengeti. PMID- 7481817 TI - Ensemble activity and behavior: what's the code? PMID- 7481818 TI - Resistance to radiation. PMID- 7481819 TI - Origin of bilaterian body plans: evolution of developmental regulatory mechanisms. AB - An argument is proposed to explain the origin of large metazoans, based on the regulatory processes that underlie the morphogenetic organization of pattern in modern animals. Genetic regulatory systems similar to those used in modern, indirectly developing marine invertebrates are considered to indicate the Precambrian regulatory platform on which were erected innovations that underlie the development of macroscopic body plans. Those systems are genetic regulatory programs that produce groups of unspecified "set-aside cells" and hierarchical regulatory programs that initially define regions of morphogenetic space in terms of domains of transcription factor expression. These ideas affect interpretation of the development of arthropods and chordates as well as interpretation of the role of the genes of the homeotic complex in embryogenesis. PMID- 7481820 TI - Opposing effects of ERK and JNK-p38 MAP kinases on apoptosis. AB - Apoptosis plays an important role during neuronal development, and defects in apoptosis may underlie various neurodegenerative disorders. To characterize molecular mechanisms that regulate neuronal apoptosis, the contributions to cell death of mitogen-activated protein (MAP) kinase family members, including ERK (extracellular signal-regulated kinase), JNK (c-JUN NH2-terminal protein kinase), and p38, were examined after withdrawal of nerve growth factor (NGF) from rat PC 12 pheochromocytoma cells. NGF withdrawal led to sustained activation of the JNK and p38 enzymes and inhibition of ERKs. The effects of dominant-interfering or constitutively activated forms of various components of the JNK-p38 and ERK signaling pathways demonstrated that activation of JNK and p38 and concurrent inhibition of ERK are critical for induction of apoptosis in these cells. Therefore, the dynamic balance between growth factor-activated ERK and stress activated JNK-p38 pathways may be important in determining whether a cell survives or undergoes apoptosis. PMID- 7481821 TI - Designer cytokines: targeting actions to cells of choice. AB - Some growth factors are therapeutically useful partly because restricted expression of their receptors limits their action to particular cell types. However, no unique stimulatory factor is known for many clinically relevant cell types, such as CD34+ hematopoietic stem cells. Here, soluble alpha receptor (R alpha) components for interleukin-6 (IL-6) and ciliary neurotrophic factor (CNTF) were targeted in an active form to cells expressing surface markers such as CD34 or CD45, thereby rendering those cells responsive to IL-6 or CNTF. The targeting of R alpha components may provide the means to create "designer" cytokines that activate a desired cell type expressing a specific cell surface marker. PMID- 7481823 TI - Self-release of CLIP in peptide loading of HLA-DR molecules. AB - The assembly and transport of major histocompatibility complex (MHC) class II molecules require interaction with the invariant chain. A fragment of the invariant chain, CLIP, occupies the peptide-binding groove of the class II molecule. At endosomal pH, the binding of CLIP to human MHC class II HLA-DR molecules was counteracted by its amino-terminal segment (residues 81 to 89), which facilitated rapid release. The CLIP (81-89) fragment also catalyzed the release of CLIP(90-105) and a subset of other self-peptides, probably by transient interaction with an effector site outside the groove. Thus, CLIP may facilitate peptide loading through an allosteric release mechanism. PMID- 7481825 TI - The development of crustacean limbs and the evolution of arthropods. AB - Arthropods exhibit great diversity in the position, number, morphology, and function of their limbs. The evolutionary relations among limb types and among the arthropod groups that bear them (insects, crustaceans, myriapods, and chelicerates) are controversial. Here, the use of molecular probes, including an antibody to proteins encoded by arthropod and vertebrate Distal-less (Dll and Dlx) genes, provided evidence that common genetic mechanisms underlie the development of all arthropod limbs and their branches and that all arthropods derive from a common ancestor. However, differences between crustacean and insect body plans were found to correlate with differences in the deployment of particular homeotic genes and in the ways that these genes regulate limb development. PMID- 7481824 TI - Cytotoxic T lymphocyte lysis inhibited by viable HIV mutants. AB - Immune evasion by the human immunodeficiency virus (HIV) is unexplained but may involve the mutation of viral antigens. When cytotoxic T lymphocytes engaged CD4 positive cells that were acutely infected with HIV bearing natural variant epitopes in reverse transcriptase, substantial inhibition of specific antiviral lysis was observed. Mutant viruses capable of these transactive effects could facilitate the persistence of a broad range of HIV variants in the face of an active and specific immune response. PMID- 7481822 TI - Sequence and characterization of a coactivator for the steroid hormone receptor superfamily. AB - A yeast two-hybrid system was used to identify a protein that interacts with and enhances the human progesterone receptor (hPR) transcriptional activity without altering the basal activity of the promoter. Because the protein stimulated transactivation of all the steroid receptors tested, it has been termed steroid receptor coactivator-1 (SRC-1). Coexpression of SRC-1 reversed the ability of the estrogen receptor to squelch activation by hPR. Also, the amino terminal truncated form of SRC-1 acted as a dominant-negative repressor. Together, these results indicate that SRC-1 encodes a coactivator that is required for full transcriptional activity of the steroid receptor superfamily. PMID- 7481826 TI - Selective opioid inhibition of small nociceptive neurons. AB - Opioid analgesia, the selective suppression of pain without effects on other sensations, also distinguishes between different types of pain: severe, persistent pain is potently inhibited by opioids, but they fail to cohceal the sensation of a pinprick. The cellular basis for this specificity was analyzed by means of patch-clamp experiments performed on fluorescently labeled nociceptive neurons (nociceptors) that innervate rat tooth pulp. Activation of the mu opioid receptor inhibited calcium channels on almost all small nociceptors but had minimal effect on large nociceptors. Somatostatin had the opposite specificity, preferentially inhibiting calcium channels on the large cells. Because persistent pain is mediated by slow-conducting, small nociceptors, opioids are thus likely to inhibit neurotransmitter release only at those primary synapses specialized for persistent pain. PMID- 7481827 TI - Regulated expression of the neural cell adhesion molecule L1 by specific patterns of neural impulses. AB - Development of the mammalian nervous system is regulated by neural impulse activity, but the molecular mechanisms are not well understood. If cell recognition molecules [for example, L1 and the neural cell adhesion molecule (NCAM)] were influenced by specific patterns of impulse activity, cell-cell interactions controlling nervous system structure could be regulated by nervous system function at critical stages of development. Low-frequency electrical pulses delivered to mouse sensory neurons in culture (0.1 hertz for 5 days) down regulated expression of L1 messenger RNA and protein (but not NCAM). Fasciculation of neurites, adhesion of neuroblastoma cells, and the number of Schwann cells on neurites was reduced after 0.1-hertz stimulation, but higher frequencies or stimulation after synaptogenesis were without effect. PMID- 7481828 TI - Scope of the AIDS epidemic in the United States. AB - Two-dimensional deconvolution techniques are used here to reconstruct age specific human immunodeficiency virus (HIV) infection rates in the United States from surveillance data on acquired immunodeficiency syndrome (AIDS). This approach suggests that 630,000 to 897,000 adults and adolescents in the United States were living with HIV infection as of January 1993, including 107,000 to 150,000 women. The estimated incidence of HIV infection declined markedly over time among white males, especially those older than 30 years. In contrast, HIV incidence appears to have remained relatively constant among women and minorities. As of January 1993, prevalence was highest among young adults in their late twenties and thirties and among minorities. An estimated 3 percent of black men and 1 percent of black women in their thirties were living with HIV infection as of that date. If infection rates remain at these levels, HIV must be considered as endemic in the United States. PMID- 7481829 TI - [Postoperative homeostatic imbalance after trauma surgical interventions of various degrees in polytrauma]. AB - The objective of this study was to evaluate and compare the derangement of body homeostatis and the inflammatory response after different types of traumatological operations in patients with multiple injuries. These were determined in a total of 60 operations. The procedures comprised osteosynthesis of the femur (n = 28), the pelvic girdle (n = 11) the spine (n = 8), and facial and basal skull reconstructions (n = 13). Specific and unspecific parameters of the inflammatory response were determined on the morning of the operation, immediately after the procedure, every 6 h on the 1st day and 48 h after the end of surgery. After all types of operations (pelvis, femur, spine, face/basal skull) significant alterations were observed for neutrophil elastase, C-reactive protein, interleukin 6, interleukin 8, antithrombin III, partial thromboplastin time and other parameters. The degree of postoperative changes differed significantly (Kruskal-Wallis test, P < 0.05) among the four types of operations for lactate, heart rate, PO2/FiO2 ratio and nitrogen excretion and showed a strong discriminating tendency for neutrophil elastase and C-reactive protein. The changes were most pronounced after operations on the pelvic girdle, followed by procedures in the femoral, spinal, and facial/basal skull regions. We conclude that a considerable inflammatory response and pronounced disturbance of body homeostasis follow traumatological operative procedures, varying in severity with the type of surgery. Several parameters allow quantitation of the surgical trauma and differentiation between different operations/regions. Further research should focus on the interrelationship between pre-existing preoperative inflammation and the additional trauma inflicted by surgery in patients with severe injuries.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7481830 TI - [Peripheral vascular injuries in polytrauma]. AB - Between 1972 und 1993 a total of 68 patients were treated at the Department of Surgery of the University Clinic of Mannheim for peripheral vascular injury resulting from multiple trauma. The average age of these patients was 31.3 years, and most of them were male (88.2%; n = 60). The injured vessels were localized evenly in all the extremities: 31 patients (45.5%) presented with arterial damage of the upper extremity, and 37 (54.5%) showed lesions along the femoro-popliteal arteries. The most frequent location of injured vessels in the multiply traumatized patient was the popliteal artery (n = 18, 26.5%), the distal part of the superficial femoral artery (n = 12, 17.6%), the brachial artery (n = 14, 20.6%) and the axillary artery (n = 10, 14.6%). The dominant cause, of trauma was road traffic accidents (72%), and 20 patients (29%) acquired their vascular injuries as motorcyclists. There were also 13 occupational accidents (19%) involving vascular injuries. In addition to a vascular trauma 34 patients (50%) had complicated fractures, and a further 34 patients (50%) had multiple fractures: 12 (17.6%) had head and brain damage, 5 (7.3%) had blunt abdominal trauma and 6 (8.8%) had blunt thoracic injury. The general amputation rate was 2.9% (n = 2). One patient died on the table of a torn off subclavian artery combined with multiple other injuries. Paresis of the plexus is a particular problem after vascular lesions of the upper extremity: in 22 patients (71%) paresis of the plexus persisted after successful vascular reconstruction (follow up period between 3 months and 16 years, median time 3.45 years).(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7481831 TI - [Surgical therapy of Achilles tendon rupture]. AB - From 1980 to 1991, 314 patients with acute rupture of the Achilles tendon were treated operatively in the "Berufsgenossenschaftliche Unfallklinik Tubingen" with suture of the tendon combined with a triceps surae tendon tip-over graft and subsequent cast immobilization for 6 weeks. Analysis of these cases revealed a significant reduction of severe soft tissue complications (3.2%), which were all controllable and a very low rerupture rate (0.3%) compared with the results obtained in the relevant literature. In 223 of these patients postoperative assessment according to the scale of Holz was carried out an average of 8.1 years after injury. The results were assessed as good in 87.4%, satisfactory in 11.2% and poor in 1.4%. Compared with conservative treatment with immobilization these results are significantly better than reported in the literature. The number of patients with Achilles tendon rupture who have received conservative functional treatment with a newly developed boot is still so small that reliable statistical statements are not yet possible. Therefore, operative therapy with an established procedure should still be regarded as the method of choice for treatment of Achilles tendon rupture; the tip-over graft is a valuable complement to the tendon suture. PMID- 7481832 TI - [Reduction of the anterior drawer of the knee joint by rehabilitation orthoses. Comparison of the MVP orthosis vs. the Donjoy-Gold point orthosis]. AB - In 20 patients with chronic anterior instability of one knee, two rehabilitation braces (MVP, Donjoy-Gold point) were tested for their ability to limit tibiofemoral translation. A KT-1000 arthrometer was used to quantify the anterior drawer. Both braces significantly reduced passive anterior drawer at 67 N, 89 N and maximal manual shear load and also active anterior drawer. Without a brace the mean anterior drawer was 12.3 mm (67 N), 14.4 mm (89 N), 17.2 mm (maximal manual drawer) and 9.1 mm (active drawer). With the MVP brace the mean anterior drawer was 6.8 mm (67 N), 8.5 mm (89 N), 11.0 mm (maximal manual drawer) and 6.8 mm (active drawer). With the Donjoy brace the mean anterior drawer was 9.2 mm (67 N), 11.0 mm (89 N), 13.1 mm (maximal manual drawer) and 7.4 mm (active drawer). Control values for the non-injured opposite knee were 6.5 mm (67 N), 7.2 mm (89 N), 8.1 mm (maximal manual drawer) and 5.6 mm (active drawer). In direct comparison, the MVP brace provided better stabilization than the Donjoy-Gold point brace did. PMID- 7481833 TI - [Covered osteosynthesis through cannulated screws in medial Garden type I and II femoral neck fractures]. AB - Fracture of the femoral neck is a common problem in elderly patients. In these patients with concomitant illness and medical complications the hip fracture is then often the reason for a long-term institutionalization with serious implications for the outcome. While the treatment of displaced femoral neck fractures Garden's stage III and IV is quite clear, the procedure for impacted or undisplaced fractures Garden's stage I or II is still the subject of controversy. In a retrospective study was shown that conservative treatment of impacted or undisplaced femoral neck fractures is associated with a high complication rate of 30-40%. When these femoral neck fractures were broken down according to Garden's classification the complication rate in cases of Garden's stage II was found to be 1.5 times that in femoral neck fractures in Garden's stage I. With a P-value less than 0.05 this difference was statistically significant (Wilcoxon test). Because of these major problems associated with conservative treatment we changed our current algorithm. Covered internal fixation with cancellous screws is now considered to be indicated for all impacted or undisplaced femoral neck fractures. Between 1990 und 1992, 53 patients (42 female, 11 male) with femoral neck fracture in Garden's stage I (24 patients) or II (29 patients) were treated in our Department of Traumatology with 6.5 mm cancellous screws. In all case early mobilization and weight-bearing were allowed. In this prospective study the average follow-up was 14.3 +/- 2.1 months (range 8-20 months).(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7481834 TI - [Post-traumatic os odontoideum]. AB - This is a report about a 20-month-old child with acquired os odontoideum caused by a fall. Diagnosis of atlanto-axial instability was eventually made 29 months later after repeated trauma and transitory tetraparesis. Dorsal C1/2 fusion with an autogenic rib graft and plaster fixation for 8 weeks failed; an autogenic iliac bone graft plus external fixation for 12 weeks finally resulted in solid fusion. PMID- 7481835 TI - [Birth trauma as the cause of fracture of the distal epiphysis of the humerus. A case report]. AB - The authors report on a case of fracture of the distal humeral epiphysis sustained at birth. This is extremely rare. The main problem in treatment of this kind of fracture is correct and timely diagnosis. In almost all reported cases the fracture has been confused with elbow dislocation. The reason for this mistake is difficult roentgenological orientation owing to invisible ossification nuclei. Ultrasono und examination is recommended for diagnosis. The treatment of the injury is conservative. Closed reduction and immobilization for 3 weeks in an above-the-elbow plaster cast is advised. The end-results have been very good in all published cases, as they were in the present case, in which the shape of the injured elbow corresponded absolutely to that on the healthy side, mobility was unrestricted and no growth disturbances were observed after 9 months. PMID- 7481836 TI - [Experiences with using the soft cast in injuries of the fibular ligament of the upper ankle joint]. AB - From June 1993 until October 1994, 75 persons with recent ruptures in the region of the fibular ligament at the ankle joint were treated with a Softcast at our hospital. The patients were divided into a surgical and a conservative group: 33 patients were operated on and 42 were treated conservatively; 49 persons were male, 26 female. Both groups were initially treated with a conventional cast, and after the first week a Softcast was used. Surgery was performed whenever X-ray examination showed a difference of more than 10 degrees in supinatory subluxation between the unstable ankle and the intact one or anterior instability of more than 10 mm. In 55 of these patients follow up by clinical and X-ray examination was possible after an average period of 14.5 months after trauma and treatment. For the clinical follow-up the 100-point Pforringer-Stolz score was used (6), and the X-ray examination was performed under constant conditions by means of these standardized conditions, excellent and good results were achieved in more than 95% of the patients. The authors recommend the semi-rigid immobilization with Softcast in both operatively and conservatively treated patients with ruptures of the lateral ankle ligaments. PMID- 7481837 TI - [Current status of diagnosis and therapy of chronic collateral ligament instability of the ankle joint: results of a survey of 267 German clinics in 1994]. AB - A questionnaire was mailed to the heads of 400 German orthopaedic departments listed by the "Deutsche Gesellschaft fur Unfallchirurgie", with the aim of evaluating the diagnostic and therapeutic approaches to chronic ankle instability that were current in 1994. The questionnaire was structured to focus on diagnostic imaging techniques, conservative and surgical management, and postoperative treatment. Evaluable questionnaires were returned from 267 hospitals (66.7%). The number of ligament reconstructions performed per institution ranged from 1 to over 100 (mean 17.5). Radiographic stress investigations and clinical examination were considered by 71.6% of the surgeons to be the most important criteria for surgical therapy. The methods most frequently used for primary reconstruction of the ligaments were: periosteal flaps (45.3%), anatomical repair (23.6%), different tenodeses (20.6%), and free anatomical implants of autologous or exogenous material (10.5%). Half the respondents used reconstruction procedures involving secondary repair of the injured ligament without augmentation or with periosteal flaps. The other half of the German surgeons preferred tenodeses or various other procedures. Conservative management of chronic ankle instability was considered by surgeons to be less appropriate for young active patients. Postoperative treatment includes casting and orthosis. The duration varied from 2 weeks to 8 weeks. The results of the nationwide survey demonstrate that more objective clinical methods of diagnosis are needed to allow preoperative differentiation between functional and mechanical instability. With better knowledge of the function of individual ligaments and more frequent exposure of the ligaments for simple overlap and direct suture, a lower rate of primary non-anatomical reconstructions may be possible in the future. PMID- 7481838 TI - Gemcitabine--status of preclinical studies and perspectives for future clinical applications of this novel nucleoside analog. Symposium proceedings. Amsterdam, The Netherlands, March 15, 1994. PMID- 7481839 TI - Gemcitabine: a modulator of intracellular nucleotide and deoxynucleotide metabolism. AB - Gemcitabine (2',2'-difluorodeoxycytidine, dFdC) is a deoxycytidine (dCyd) analog that extensively modulates intracellular CTP and dCTP metabolism. In Chinese hamster ovary (CHO) cells, a 4-hour exposure to gemcitabine (100 mumol/L) reduced cellular CTP and dCTP concentrations to 5.9% and 50%, respectively. Intracellular UTP concentrations increased, indicating a metabolic block at CTP synthetase. Pool-sizes of ATP and GTP remained unaffected. In contrast, a CHO mutant deficient in deoxycytidine kinase, and thus unable to accumulate dFdCTP, maintained its CTP pools under identical conditions, suggesting that the CTP pool depletion was dependent on dFdC phosphorylation. Neither 100 mumol/L arabinosylcytosine nor 5 mmol/L hydroxyurea affected CTP levels, indicating that inhibition of DNA synthesis by analog incorporation or by depletion of dNTP pools were not the causes of the CTP pool perturbation. Metabolic studies demonstrated that incorporation of [3H]uridine into the UTP pool was not impaired by dFdC treatment, whereas the specific activity of the CTP pools decreased as a function of increasing gemcitabine concentration and time of exposure. Comparable results were obtained using 3-deazauridine, a known inhibitor of CTP synthetase. We conclude that high cellular concentrations of dFdCTP deplete cellular CTP concentrations by inhibition of the dCTP pool and also may be a limiting factor for RNA synthesis. PMID- 7481840 TI - Induction of apoptosis by gemcitabine. AB - Inhibition of cellular DNA synthesis is the major action of gemcitabine. In cells, this drug is converted to its triphosphate (dFdCTP), which is incorporated into DNA and terminates DNA strand elongation. After incorporation of gemcitabine nucleotide into the DNA strand, one more deoxynucleotide is incorporated, and thereafter the DNA polymerases are unable to proceed ("masked chain termination"). Gemcitabine also inhibits DNA synthesis indirectly by decreasing cellular dNTP pools via inhibition of ribonucleotide reductase. Incubation of human leukemia cells (CEM) with gemcitabine leads to apoptotic cell death. Two types of DNA fragmentation were observed in the gemcitabine-treated cells: (1) large-sized double-stranded DNA fragments range from 5 kb to 500 kb with the majority of the fragments located at 50 kb, and (2) nucleosomal-sized DNA fragments. Both types of drug-induced DNA fragmentation were detected in exponentially growing cells and were much more prominent in cells synchronized at S phase. The gemcitabine-induced DNA fragmentation in either synchronized or nonsynchronized cells was inhibited by the DNA synthesis inhibitor, aphidicolin. Thus, incorporation of gemcitabine into DNA is essential to induce DNA fragmentation. The intracellular calcium chelator BAPTA-AM inhibited the drug induced nucleosomal DNA fragmentation but did not prevent the large-sized DNA fragmentation, suggesting that the nucleosomal DNA fragmentation is a calcium dependent event, whereas the large-sized DNA fragmentation is independent of calcium. Furthermore, BAPTA-AM did not prevent the morphologic appearance of apoptotic bodies in cells incubated with gemcitabine, indicating that degradation of DNA to nucleosomal fragments is not an essential element of the apoptotic process. Phorbol 12-myristate 13-acetate also inhibited drug-induced nucleosomal DNA fragmentation, but prevented neither large-sized DNA fragmentation nor formation of apoptotic bodies. In contrast, aphidicolin inhibited both types of DNA fragmentation and blocked the formation of apoptotic bodies in the presence of gemcitabine. These data suggest that the generation of large-sized DNA fragments caused by incorporated gemcitabine monophosphate in DNA is critical in gemcitabine-induced apoptosis, whereas nucleosomal DNA fragmentation is not a requirement in this cell death process. PMID- 7481841 TI - Consequences of 2',2'-difluorodeoxycytidine (gemcitabine) on replicative DNA synthesis in intact HL-60 cells. AB - The technique of pH-step alkaline elution was used to assess the effects of gemcitabine (dFdC) on replicative DNA synthesis in intact HL-60 human myeloid leukemia cells. Although gemcitabine did cause profound inhibition of DNA chain elongation, it was progressively incorporated through nascent DNA replication intermediates of increasing size into genomic DNA. Hence, in the intact cell, it is not a chain terminator, at least not in the absolute sense of the term. In comparison to cytosine arabinoside (ara-C), the progression of incorporated gemcitabine from small nascent DNA fragments to genomic-length DNA was less complete. Furthermore, at equitoxic exposures, less gemcitabine was incorporated into DNA than ara-C. Studies of the effects of gemcitabine on ribonucleotide reduction in HL-60 cells revealed that dGTP pools, but not dCTP pools, were reduced by a 3-hour exposure to 40 nmol/L gemcitabine (the concentration that causes 50% lethality). This reduction was transient, and recovery of dGTP pool size was accomplished within 16 hours. These studies indicate that the effects of gemcitabine on inhibiting replicative DNA chain elongation comprise an important component of the cytotoxicity of the drug. PMID- 7481842 TI - Gemcitabine: metabolism, mechanisms of action, and self-potentiation. AB - Gemcitabine (dFdC) is a new anticancer nucleoside that is an analog of deoxycytidine. It is a pro-drug and, once transported into the cell, must be phosphorylated by deoxycytidine kinase to an active form. Both gemcitabine diphosphate (dFdCTP) and gemcitabine triphosphate (dFdCTP) inhibit processes required for DNA synthesis. Incorporation of dFdCTP into DNA is most likely the major mechanism by which gemcitabine causes cell death. After incorporation of gemcitabine nucleotide on the end of the elongating DNA strand, one more deoxynucleotide is added and thereafter, the DNA polymerases are unable to proceed. This action ("masked termination") apparently locks the drug into DNA as the proofreading enzymes are unable to remove gemcitabine from this position. Furthermore, the unique actions that gemcitabine metabolites exert on cellular regulatory processes serve to enhance the overall inhibitory activities on cell growth. This interaction is termed "self-potentiation" and is evidenced in very few other anticancer drugs. PMID- 7481843 TI - Induction of resistance to 2',2'-difluorodeoxycytidine in the human ovarian cancer cell line A2780. AB - 2',2'-Difluorodeoxycytidine (gemcitabine; dFdC) is a nucleoside analog with promising antitumor activity. To be active it must be phosphorylated by deoxycytidine kinase (dCK). We induced resistance to gemcitabine in the human ovarian carcinoma cell line A2780 by exposure to increasing concentrations of gemcitabine. At 72 hours' exposure the IC50, defined as the concentration of gemcitabine causing 50% growth inhibition, increased from 0.6 nmol/L gemcitabine in A2780 to 92 mumol/L in the resistant variant, AG6000. AG6000 is cross resistant to other drugs that require activation by dCK, such as I-beta-D arabinofuranosylcytosine, 5-aza-2'-deoxycytidine, and 2-chlorodeoxyadenosine. AG6000 was also cross-resistant to 2',2'-difluorodeoxyuridine (dFdU), the deamination product of gemcitabine. In addition, cross-resistance to the multidrug-resistance drugs doxorubicin and vincristine was observed. This was not associated with induction of P-glycoprotein. No accumulation of gemcitabine triphosphate could be detected in AG6000 cells, in contrast to the parental A2780 cells. There was no specific dCK activity in extracts from AG6000 cells. Western blot analysis using a polyclonal anti-dCK antibody did not reveal any dCK protein in AG6000 cell extracts. Reverse transcribed and polymerase chain reaction amplified mRNA, using specific dCK primers, demonstrated that AG6000 expressed a normal length amplicon of 701 base pairs, besides an aberrant amplicon of 500 base pairs. Although the resistant cell line is routinely cultured in 6 mumol/L gemcitabine, the resistant phenotype can be maintained for at least 10 passages without gemcitabine. These results indicate that the gemcitabine resistance phenotype is stable and mainly due to dCK deficiency. PMID- 7481844 TI - Schedule-dependent antitumor effect of gemcitabine in in vivo model system. AB - Gemcitabine (2',2'-difluorodeoxycytidine, dFdC, LY188011) is a new deoxycytidine analog with preclinical antitumor activity in solid tumors from murine and human origin. Of particular importance is the fact that the therapeutic effects of gemcitabine at the maximum tolerated dose level are dependent on the administration schedule. This paper describes the sensitivity pattern of gemcitabine in human head and neck squamous cell carcinoma, ovarian carcinoma, and soft tissue sarcoma, all growing as xenografts in athymic nude mice. The drug was injected intraperitoneally in various schedules at equitoxic, maximum tolerated dose levels, resulting in a reversible weight loss that varied between 5% and 15%. Generally, it was found that treatment with 120 mg/kg gemcitabine, injected four times at 3-day intervals, was more effective than the schedules of daily (five times 2.5 to 3.5 mg/kg) or weekly (two times 240 mg/kg) injections. Other workers have shown that this 3-day interval schedule also was active in human pancreas and lung carcinoma xenografts. Additional experiments were performed on normal mice bearing the colon 26-10 murine colon carcinoma. The effect of a continuous intravenous infusion system was investigated by giving two injections of 15 mg/kg gemcitabine for 24 hours at a 7-day interval. Interestingly, the efficacy of treatment increased dramatically with this infusion schedule, producing complete remissions in most tumors. In conclusion, our data on the effect of gemcitabine in animal tumor models indicate that (1) the time interval between push injections is important when intermittent schedules are used and (2) continuous infusions over a 24-hour period can be very effective in in vivo models. PMID- 7481845 TI - Evaluation of the cytotoxic activity of gemcitabine in primary cultures of tumor cells from patients with hematologic or solid tumors. AB - Gemcitabine (2'-deoxy-2',2'-difluorocytidine; dFdC) is a novel nucleoside analog that has shown clinical activity against solid tumors. The semiautomated fluorometric microculture cytotoxicity assay (FMCA) was used for evaluation of the cytotoxicity of gemcitabine in vitro in primary cultures of human cells from patients with hematologic or solid tumors. The activity pattern of cytarabine (ara-C), 2-chlorodeoxyadenosine (CdA), etoposide (VP-16), doxorubicin, and cisplatin were included for comparison. One hundred forty samples were tested using continuous drug exposure. Gemcitabine showed high activity against hematologic samples, whereas little or no activity was observed in the solid tumor groups. A similar pattern of activity also was observed for ara-C and CdA, whereas etoposide, cisplatin, and doxorubicin were relatively more active in solid tumors. Cross-resistance analysis between gemcitabine and the standard drugs revealed the following rank order of correlation coefficients: ara-C > doxorubicin > CdA > cisplatin > VP-16. The results indicate that gemcitabine is differentially active against hematologic tumors and that the activity pattern of gemcitabine resembles that of ara-C. However, these results also indicate that gemcitabine may be more active against some solid tumor groups in comparison to ara-C and CdA. PMID- 7481846 TI - Medicinal chemistry of difluoropurines. AB - A series of over 70 difluoropurine analogs was synthesized by varying the C-2, 6 and 8 substituents about the purine ring system. After initial in vitro and in vivo screening, testing concentrated on the 2,6-diaminopurine analog (dFdAP) and the guanosine analog (dFdG). dFDAP appears to be a prodrug for dFdG. Both compounds significantly inhibited mammary tumor growth in mice, caused a moderate inhibition in ovarian and lymphosarcoma models, and demonstrated no activity in lung and melanoma models. This is a narrower spectrum of activity than that of gemcitabine (dFdC). The antitumor activity of dFdAP in human xenografts that are refractory to standard clinical agents was comparable or superior to that of gemcitabine. However, during the preliminary toxicology testing, dFdG was associated with several deaths caused by cardiac toxicity. Therefore, although dFdG is a potentially useful oncolytic, further investigation is required. PMID- 7481848 TI - Radiosensitization of human tumor cells by gemcitabine in vitro. AB - Gemcitabine is a novel nucleoside analog that has shown clinical activity in a variety of solid tumors. The use of gemcitabine in combination with radiation was investigated in vitro using HT-29 colon carcinoma cells. Gemcitabine was demonstrated to be a potent radiosensitizer even at noncytotoxic concentrations. Increasing the concentration of gemcitabine and/or the duration of exposure increased the radiosensitization of cells. Biochemical studies indicated that a reduction in the dATP pool was the most likely mechanism of radiosensitization by gemcitabine. This effect is of great clinical importance and should be investigated in vivo. PMID- 7481847 TI - Difluorodeoxyguanosine: cytotoxicity, metabolism, and actions on DNA synthesis in human leukemia cells. AB - The success of gemcitabine (2',2'-difluorodeoxycytidine; dFdC) resulted in new interest in its purine congeners. Based on the structure-activity relationship studies of catabolism and anabolism, 2',2'-difluorodeoxyguanosine (dFdG) emerged as a lead candidate among the difluoropurine analogs. The cytotoxicity, metabolism, and actions of dFdG on DNA synthesis were studied in the human leukemia lymphoblastoid line CCRF-CEM. The IC50 values of dFdG after a 72-hour continuous incubation were 0.01, 0.03, and 0.28 mumol/L for CCRF-CEM, K562, and HL-60 cells, respectively. A cell line deficient in dCyd kinase was equally sensitive to dFdG, suggesting that, in contrast to dFdC, dFdG may be activated by other deoxynucleoside kinase(s). Consistent with these data, coincubation with dGuo spared the dFdG-mediated toxicity; however, up to 500 mumol/L dCyd failed to reverse the toxicity of dFdG. These observations indicated that dGuo kinase, which phosphorylates arabinosylguanine, also appears to play a major role in activating dFdG. CCRF-CEM cells incubated with varying concentrations of [3H]dFdG accumulated dFdGTP in a dose-dependent manner; a 3-hour incubation with 1 mmol/L dFdG resulted in more than 600 mumol/L intracellular dFdGTP. This is in contrast to the gemcitabine triphosphate accumulation, which is saturated at 10 to 20 mumol/L of exogenous dFdC. dFdG metabolites affected ribonucleotide reductase, resulting in a lowering of the dCTP pool; this is in agreement with the effect of dFdC on dNTP pools in leukemia cell lines. The major effect of dFdG on macromolecular synthesis was inhibition of DNA synthesis. DNA primer extension over a defined template revealed that dFdGTP was a good substrate for DNA polymerase alpha and incorporated opposite C sites of the template. Unlike arabinosyl analogs, but similar to gemcitabine triphosphate, dFdGTP incorporation caused DNA polymerase to pause after one normal deoxynucleotide was incorporated beyond the analog. The unique activation requirements of dFdG, its novel mode of inhibition of DNA synthesis, and its potent toxicity to human leukemia cells make it a promising new antimetabolite. PMID- 7481850 TI - Pharmacokinetics of paclitaxel and carboplatin in combination. AB - We studied the pharmacokinetics of paclitaxel (Taxol; Bristol-Myers Squibb Company, Princeton, NJ) and carboplatin administered in combination to 21 patients with advanced non-small cell lung cancer. Paclitaxel was administered as a 24-hour intravenous infusion at doses of 135 to 200 mg/m2. Carboplatin, dosed to a target area under the concentration-time curve of 5, 7, 9, or 11 mg/mL.min, was administered as a 20-minute infusion immediately following paclitaxel. Neither the paclitaxel concentrations at the end of the infusion nor the terminal elimination of paclitaxel, as assessed by the duration of time that plasma paclitaxel concentrations were 0.05 mumol/L or greater, were different compared with historical data of paclitaxel as a single agent. Thus, we concluded that carboplatin had no perceived effect on the pharmacokinetics of paclitaxel in this schedule. The observed areas under the concentration-time curves for carboplatin were consistently 10% to 15% less than the target values. Although this may indicate a possible interaction between paclitaxel and carboplatin, it also may have been a result of inadequate assessment of glomerular filtration rate, which was used to determine the carboplatin dose. PMID- 7481851 TI - Management of breast cancer: status and future trends. AB - Metastatic breast cancer continues to be a challenge for the clinician. Heterogeneity of natural history and drug resistance have interfered with progress in treatment. Hormonal therapy is effective in 30% of patients, some of whom derive long-term benefit from sequential hormonal manipulations. Most patients have hormone-refractory tumors, and eventually all breast cancers become hormone resistant. Cytotoxic chemotherapy is the treatment of choice for these patients. Promising leads in recent clinical investigation include the development of dose-intensive therapies for induction or remission consolidation, biochemical modulation, development of new hormonal and cytotoxic agents, targeted therapy using immunologic or ligand-driven vehicles, and growth factor modulation. Among the encouraging clinical developments, the discovery of the taxanes represents an important step in improving the efficacy of cytotoxic therapy. Paclitaxel (Taxol; Bristol-Myers Squibb Company, Princeton, NJ) produces objective regressions in 40% to 60% of patients with untreated or minimally treated metastatic breast cancer. The efficacy of this agent persists in patients with chemotherapy-refractory tumors, including those with anthracycline resistance. New paclitaxel-based combinations, especially those with doxorubicin, cisplatin, or vinorelbine, appear promising. This agent, alone or in combination, is also under evaluation in combined-modality strategies for early and advanced primary breast cancer. PMID- 7481849 TI - Interaction between cisplatin and gemcitabine in vitro and in vivo. AB - Both gemcitabine (2',2'-difluorodeoxycytidine; dFdC) and cisplatin (cis-diammine dichloroplatinum; CDDP) are active against several solid malignancies, including ovarian cancer and head and neck squamous cell carcinoma. Because of differences in mechanisms of action and toxicity profiles, combination of the two drugs has enormous clinical potential. We studied possible synergism between the drugs: in vitro using three variants of the human ovarian cancer cell line A2780, and in vivo using gemcitabine- and cisplatin-sensitive and -resistant tumors, the head and neck cancer xenografts HNX-22B and HNX-14C and the murine syngeneic colon 26 10 tumor. In vitro, cells were cultured for 72 hours and exposed to the drugs for 1 to 72 hours; synergy was evaluated by multiple drug-effect analysis. In wild type A2780 and cisplatin-resistant ADDP cells, simultaneous exposure for 24 and 72 hours was synergistic, as well as preincubation with cisplatin for 4 hours followed by gemcitabine. Preincubation with gemcitabine for 4 hours followed by gemcitabine and cisplatin was synergistic in ADDP and A2780 cells. Cisplatin did not enhance the accumulation of gemcitabine triphosphate in A2780 and ADDP cells. Cisplatin caused a marginal decrease of the number of double strand breaks in the DNA caused by gemcitabine. In vivo, gemcitabine at the maximum tolerated dose of 100 or 120 mg/kg could be combined with cisplatin at 4 mg/kg. When injected simultaneously this resulted in at least additive anti-tumor activity in HNX-22B, but not in HNX-14C and colon 26-10 tumors. Cisplatin, injected 4 hours before or after gemcitabine, was equally active as the simultaneous schedule in HNX-22B tumors, but more toxic. In conclusion, the combination of gemcitabine and cisplatin can be synergistic in vitro and at least additive in vivo; this synergism is schedule dependent. The mechanism cannot be explained by gemcitabine triphosphate accumulation or DNA damage studies. PMID- 7481852 TI - Memorial Sloan-Kettering Cancer Center experience with paclitaxel in the treatment of breast cancer. AB - Paclitaxel (Taxol; Bristol-Myers Squibb Company, Princeton, NJ) is the most important new cytotoxic agent to be introduced for the management of breast cancer in many years. During this decade, investigators at Memorial Sloan Kettering Cancer Center have conducted multiple clinical and laboratory investigations aimed at optimally integrating this agent into therapeutic strategies for breast cancer. These studies address both single-agent and combination regimens in the metastatic and adjuvant settings. This report will review previous results, but focus on active studies and future avenues of research. PMID- 7481853 TI - Biweekly paclitaxel in the treatment of patients with metastatic breast cancer. AB - Paclitaxel (Taxol; Bristol-Myers Squibb Company, Princeton, NJ) administered by a 3-hour infusion causes a rapid decline to and recovery from the hematologic white blood cell nadir. This suggests that biweekly administration of paclitaxel alone or in combination with drugs that have limited hematologic toxicity may be possible. The first study discussed in this report tried to determine the tolerability and activity of biweekly paclitaxel administered in combination with cisplatin in patients with metastatic breast cancer. Subsequently, after an impressive response rate, a second study of biweekly paclitaxel alone was initiated to attempt to discern which activity spectrum and which toxicities were due to paclitaxel and which were due to cisplatin. Patients with metastatic breast cancer who received up to one prior adjuvant chemotherapy regimen were eligible for both studies. Paclitaxel was administered intravenously by a 3-hour infusion followed by intravenous cisplatin biweekly in the ambulatory setting. In the second study, paclitaxel was administered alone. Twenty-nine patients have been entered in the combination study, of whom 27 had received prior adjuvant therapy and 23 had received prior anthracyclines. Dose-limiting toxicity for the phase I study of paclitaxel and cisplatin proved to be a failure to recover neutrophil counts greater than 750 cells/microL by day 14. The maximum tolerated dose was paclitaxel 90 mg/m2 and cisplatin 60 mg/m2 every 2 weeks. Nonhematologic toxicities were mild and included fatigue, arthralgias, and nausea and vomiting. At this time, the 27 patients evaluable for response have achieved an 85% response rate and an 11% complete response rate. Complete responses have been seen in soft tissue, lung, and nodal disease. All patients with complete responses have had previous anthracyclines. The biweekly paclitaxel-alone study is still accruing patients. The current paclitaxel dose level is 150 mg/m2. It is still too early to evaluate response; however, response rates appear to be less impressive than those seen with combined paclitaxel/cisplatin. The final results of these studies are pending. PMID- 7481854 TI - Doxorubicin/paclitaxel combination chemotherapy for metastatic breast cancer: the Eastern Cooperative Oncology Group experience. AB - The addition of paclitaxel (Taxol; Bristol-Myers Squibb Company, Princeton, NJ) to the therapeutic armamentarium for breast cancer has resulted in novel opportunities and challenges for the clinician. In a series of trials that began in 1992, the Eastern Cooperative Oncology Group (ECOG) investigated the role of paclitaxel in combination with doxorubicin for the treatment of patients with advanced breast cancer. The design of the first trial, a limited-institution pilot study, involved alternating doxorubicin and paclitaxel as single agents. The alternating schedule was well tolerated and associated with objective responses in seven of 12 patients entered into the trial. In the second trial, a limited-institution phase I trial, bolus doxorubicin was combined with paclitaxel administered over a 24-hour infusion, with administration of the two drugs separated by 4 hours. While no dose-limiting toxicity was seen at doses of 50 and 150 mg/m2 of doxorubicin and paclitaxel, respectively, dose-limiting mucositis occurred at respective doses of 60 and 175 mg/m2. Mucositis was more common when administration of paclitaxel preceded that of doxorubicin than when doxorubicin preceded paclitaxel. Based on these results, the ECOG initiated a phase III trial comparing single-agent paclitaxel, single-agent doxorubicin, and the combination of paclitaxel and doxorubicin. This study, now an Intergroup trial of ECOG, the Southwest Oncology Group, and the North Central Cancer Treatment Group, is designed to accrue 730 patients. This trial will close to accrual in September 1995, and analysis of the trial should provide useful information regarding the potential synergy of doxorubicin and paclitaxel, the degree of cross-resistance between the two compounds, and the relationship between steady-state paclitaxel levels and response to therapy. In addition, ECOG is currently conducting a trial designed to confirm the striking activity of cisplatin and paclitaxel seen in the British Columbia trials. Future trials by the group will examine means of combining paclitaxel with other active agents. PMID- 7481855 TI - Phase I study of paclitaxel and carboplatin: implications for trials in head and neck cancer. AB - An ongoing phase I and pharmacokinetic trial of paclitaxel (Taxol; Bristol-Myers Squibb Company, Princeton, NJ) in combination with carboplatin is evaluating the maximum tolerated dose (MTD) of a 3-hour paclitaxel infusion combined with fixed doses of carboplatin in previously treated and untreated patients with a variety of advanced cancers. A patient's previous treatment status determines the fixed carboplatin dose: target area under the concentration-time curves of 4.0 and 4.5 mg.min/mL in previously treated and untreated patients, respectively. Studies 1 and 2 entered previously treated patients to establish the paclitaxel MTD without and with cytokine support: study 1 established 135 mg/m2 paclitaxel as the MTD without such support. In study 2, granulocyte colony-stimulating factor is administered, and the MTD has not yet been reached with paclitaxel doses of 135 mg/m2 to 230 mg/m2 assessed thus far and 250 mg/m2 now being evaluated. Objective responses have been seen in three of five patients with squamous cell carcinoma of the head and neck and in patients with non-small cell lung cancer and metastatic cancer of unknown primary site as well. Myelosuppression has been the dose-limiting toxicity, although significant nausea and vomiting and myalgia have been documented occasionally. Paclitaxel apparently has nonlinear pharmacokinetics with a beta half-life of 6.7 hours (SD +/- 1.3 hours). Future trials of paclitaxel/carboplatin will address the management of squamous cell carcinoma of the head and neck and non-small cell carcinoma of the lung. PMID- 7481856 TI - Feasibility and pharmacokinetics of paclitaxel, carboplatin, and concurrent radiotherapy for regionally advanced squamous cell carcinoma of the head and neck and for regionally advanced non-small cell lung cancer. AB - Sequential chemotherapy and radiotherapy offer considerable improvements in the care of patients with locally advanced non-small cell lung cancer (NSCLC) and squamous cell carcinoma of the head and neck (SCCHN). Improved survival for lung cancer and organ preservation in head and neck cancer have occurred with this approach, but local control remains a problem. Concurrent chemotherapy and radiotherapy can potentially improve both local control and control of micrometastases. We previously showed that concurrent carboplatin plus radiotherapy is a useful potential treatment for advanced NSCLC and SCCHN, producing good local control and acceptable toxicity. Paclitaxel (Taxol; Bristol Myers Squibb Company, Princeton, NJ) has recently demonstrated strong single agent activity against both NSCLC and SCCHN. Paclitaxel has also shown favorable interactions with radiotherapy and with platinum compounds. We therefore added weekly paclitaxel at 45 mg/m2 given after premedication and before carboplatin (100 mg/m2) weekly during concurrent standard-dose radiotherapy. Twenty patients (seven with SCCHN and 13 with NSCLC) have been treated (38 and 73 weekly doses, respectively). Toxicities have been manageable with delay or dose reduction in five and eight patients, respectively, for SCCHN and NSCLC. Based on these toxicities paclitaxel dose has been reduced to 40 mg/m2/wk. Plasma pharmacokinetics have shown that concurrent carboplatin and radiotherapy do not alter the pharmacokinetic behavior of paclitaxel compared with single-agent data. Concurrent therapy with carboplatin, paclitaxel, and radiotherapy is feasible on this schedule. Further case accrual to assess efficacy is ongoing. PMID- 7481857 TI - Phase I study of paclitaxel, cisplatin, and ifosfamide in patients with recurrent or metastatic squamous cell cancer of the head and neck. AB - The prognosis for patients with recurrent or metastatic squamous cell cancer of the head and neck is poor. Chemotherapy has not significantly improved survival. New agents and regimens are being developed in hopes of improving the outcome for these patients. A phase I/II trial using a combination of paclitaxel (Taxol; Bristol-Myers Squibb Company, Princeton, NJ), cisplatin, and ifosfamide is being performed. The phase I portion is completed and reported here. Granulocytopenia was the dose-limiting toxicity. Overall, the regimen was well tolerated. Major responses were observed in four of 10 evaluable patients in the phase I study. The phase II trial is ongoing. PMID- 7481858 TI - Weekly, outpatient paclitaxel and concurrent cranial irradiation in adults with brain tumors: preliminary results and promising directions. AB - In a phase I study, paclitaxel (Taxol; Bristol-Myers Squibb Company, Princeton, NJ) was given weekly by 3-hour infusion for 6 weeks concurrent with daily cranial irradiation as initial treatment for patients with newly diagnosed astrocytic brain tumors. The primary goal of the study was to establish the maximum tolerated dose of paclitaxel when administered by this schedule. Sixty patients were entered into the study and received at least one course of therapy. Fifty six patients completed treatment. The treatment regimen was well tolerated, with minimal hematologic toxicity. Peripheral neuropathy was dose-limiting. Median survival was 9.2 months for patients with glioblastoma multiforme and has not been reached for patients with anaplastic astrocytoma or astrocytoma. The maximum tolerated dose of paclitaxel in this study was 250 mg/m2 administered weekly. However, because five of six patients receiving this dose of paclitaxel developed peripheral neuropathy, and because patients with brain tumors may adapt to a superimposed neuropathy less well than patients without pre-existing nervous system dysfunction, we propose 225 mg/m2 as the recommended dose for subsequent phase II trials. PMID- 7481859 TI - The role of paclitaxel in the therapy of bladder cancer. AB - Although significant strides have been made in the development of effective chemotherapy for metastatic bladder cancer, those benefits have plateaued and prompted a search for new, active agents. The most active of a new generation of investigational agents is paclitaxel (Taxol; Bristol-Myers Squibb Company, Princeton, NJ). The only single-agent phase II trial to date, which used paclitaxel at a dose of 250 mg/m2 by 24-hour infusion every 3 weeks, resulted in a 42% response rate and a 27% complete response rate in previously untreated patients. With granulocyte colony-stimulating factor support, this therapy was well tolerated, with fewer than 10% of patients developing neurtropenic fever during therapy. Paclitaxel's potential as a radiation sensitizer and its relatively low renal excretion make it an attractive agent for use in patients with urothelial carcinoma. The goal of ongoing and planned clinical trials will be to (1) identify potential paclitaxel-containing combination regimens, (2) define the optimal dose and schedule of administration in patients with bladder cancer, and (3) define the activity of paclitaxel in previously treated patients. PMID- 7481860 TI - Paclitaxel plus estramustine in metastatic hormone-refractory prostate cancer. AB - Combination antimicrotubule therapy with estramustine phosphate (EMP) and vinblastine has reproducible activity in metastatic hormone-refractory prostate cancer (HRPC) with an objective response rate of 31%. Although paclitaxel (Taxol; Bristol-Myers Squibb Company, Princeton, NJ) by 24-hour infusion was inactive in HRPC, 0.5 to 1.0 nmol/L concentrations of paclitaxel combined with EMP exerted synergistic cytotoxicity in DU-145 androgen-independent human prostate cancer cell lines. Based on these results, we treated 24 patients with HRPC using the combination of paclitaxel 120 to 140 mg/m2 by 96-hour intravenous infusion every 3 weeks plus daily oral EMP at 600 mg/m2/d. Of seven patients with measurable soft tissue metastases, three have attained partial responses and a fourth patient is nearing partial response status. Of 16 patients with bone-only disease evaluated by change in serum prostate-specific antigen levels, 11 patients (68.8%) have had decreases of > or = 50% from pretreatment baseline. The prostate specific antigen decrease has exceeded 80% in six of 16 (37.5%) patients. For all 23 evaluable patients, the prostate-specific antigen has decreased by > or = 50% in 15 (65.2%) and by > or = 80% in eight (34.7%). Grade 4 leukopenia occurred in one of 21 patients treated at the paclitaxel dose of 120 mg/m2/96 hr and one of three patients treated at 140 mg/m2/96 hr. The incidence of nausea (50%) and peripheral edema (37.5%) was similar to that associated with single-agent EMP. These results demonstrate that 96-hour paclitaxel plus EMP is active in HRPC and provide further evidence that the rational combination of antimicrotubule agents leads to synergistic antitumor activity in HRPC. PMID- 7481861 TI - The activity of paclitaxel in gastrointestinal tumors. AB - Gastrointestinal malignancies, which are common around the world, are relatively refractory to available cancer chemotherapeutic agents, necessitating a search for new agents able to improve palliation and survival of patients with advanced disease. Currently, metastatic or local-regional unresectable carcinoma of the esophagus or gastroesophageal junction carries a dismal prognosis. Paclitaxel (Taxol; Bristol-Myers Squibb Company, Princeton, NJ), a new mitotic spindle inhibitor, has been studied in patients with advanced gastrointestinal carcinoma. In this phase II National Cancer Institute-sponsored study, previously untreated patients with unresectable local-regional or metastatic carcinoma of the esophagus or gastroesophageal junction (either squamous cell carcinoma or adenocarcinoma) received a starting dose of paclitaxel of 250 mg/m2 administered by a 24-hour intravenous infusion (with premedication) repeated every 21 days; all patients received subcutaneous granulocyte colony-stimulating factor 5 micrograms/kg daily 24 hours after the completion of the paclitaxel infusion. Fifty-one of 53 patients were assessable for response and response duration. Thirty-three patients had adenocarcinoma and 18 had squamous cell carcinoma. Sixteen (31%) patients achieved a response (one complete and 15 partial) and 11 (22%) achieved a minor response. Among 33 patients with adenocarcinoma, 12 (36%; 95% confidence interval, 14% to 58%) achieved either a complete (one patient) or partial (11 patients) response and six patients (18%) had a minor response. Four (22%; 95% confidence interval, 3% to 41%) of 18 patients with squamous cell carcinoma had a partial response and four (22%) had a minor response. At a median follow-up of 12+ months, 28 patients remain alive with an actuarial median survival duration of 10.2 months (range, 2 to 20+ months). These data suggest that paclitaxel is active against adenocarcinoma as well as squamous cell carcinoma of the esophagus. In a subsequent study, the combination of paclitaxel (175 mg/m2 over 3 hours on day 1), cisplatin (20 mg/m2 on days 1 to 5), and 5 fluorouracil (1,000 mg/m2/d in the first 10 patients but then reduced to 750 mg/m2/d, given as a continuous infusion on days 1 to 5) repeated every 28 days was given to patients with advanced adenocarcinoma or squamous cell carcinoma of the esophagus. Of 46 patients accrued (target accrual, 60), 35 are men and 11 are women, 30 have adenocarcinoma and 16 have squamous cell carcinoma. Among 39 patients evaluated for response so far, one has had a complete response and 16 have had partial responses (overall response rate, 44%; 95% confidence interval 28% to 59%). Five patients have had a minor response. The median granulocyte nadir was 1,200/microL.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 7481862 TI - A phase I study of cisplatin, etoposide, and paclitaxel in small cell lung cancer: a University of Colorado Cancer Center study. AB - The University of Colorado Cancer Center is conducting a phase I study of the three-drug PET combination of cisplatin, etoposide, and paclitaxel (Taxol; Bristol-Myers Squibb Company, Princeton, NJ) in patients with advanced (stage IV or IIIB with pleural effusion) small cell lung cancer. The primary study goal was to define the maximally tolerated doses given on an outpatient basis. Secondary goals were to determine toxicities, response rate, response duration, and survival. Paclitaxel was given as a 3-hour intravenous (IV) infusion prior to cisplatin and etoposide. The starting doses were paclitaxel 135 mg/m2 day I, cisplatin 80 mg/m2 IV day I, and etoposide 50 mg/m2 IV day I, and 100 mg/m2 orally days 2 and 3, every 3 weeks. In the second group, the etoposide was increased to 80 mg/m2 IV day I and 160 mg/m2 orally days 2 and 3. In the third group, paclitaxel was increased to 175 mg/m2 IV day I. Granulocyte colony stimulating factor was not given on the first cycle, but was given on subsequent cycles if grade 4 neutropenia developed. So far, 13 patients have been entered on the study; all are evaluable for toxicity and nine are evaluable for response. The major toxicity was neutropenia, with no other grade 4 toxicities observed. All patients received the full six cycles of therapy. Thus far, partial responses have been observed in four patients (44%) and complete responses in five patients (56%), for an overall response rate of 100%. This ongoing study has shown that full doses of each of these three active drugs can be administered safely on an outpatient basis. The encouraging early results should lead to a multicenter phase II evaluation of the PET combination. PMID- 7481863 TI - Current status of chemotherapy for ovarian cancer. AB - Standard treatment for patients with advanced ovarian cancer has been cytoreductive surgery followed by combination chemotherapy. Until recently, platinum-based chemotherapy was considered optimal and patients were treated with regimens built around either cisplatin or carboplatin. Recently, paclitaxel (Taxol; Bristol-Myers Squibb Company, Princeton, NJ) has been shown to be a highly active agent in refractory ovarian cancer patients. Subsequently, the Gynecologic Oncology Group performed a prospective randomized trial of paclitaxel plus cisplatin compared with cisplatin plus cyclophosphamide in suboptimal stage III and IV ovarian cancer patients. Based on higher response rates, longer time to progression, and marked improvement in median survival (37.5 months compared with 24.4 months), the Gynecologic Oncology Group currently considers paclitaxel plus cisplatin to be the new standard regimen for patients with advanced disease. More recently, paclitaxel plus carboplatin also has been evaluated in previously untreated patients. Using area under the curve dosing for carboplatin, it was demonstrated that this agent could be combined with paclitaxel (175 mg/m2 in a 3 hour infusion) with acceptable toxicity. All current Gynecologic Oncology Group protocols for untreated patients with ovarian cancer use a paclitaxel-based regimen. These clinical trials are evaluating the relative efficacy of carboplatin plus paclitaxel versus cisplatin plus paclitaxel as well as differences in dose and schedule and number of cycles of treatment. Investigational studies are continuing with high-dose chemotherapy that requires hematologic support as well as with intraperitoneal therapy (cisplatin or paclitaxel). PMID- 7481864 TI - The platinum compounds and paclitaxel in the management of carcinomas of the endometrium and uterine cervix. AB - Endometrial carcinoma and squamous cell carcinoma of the cervix are common invasive neoplasms of the female genital tract. Early diagnosis of a majority of patients has resulted in high cure rates for both diseases. In the last two decades, a growing number of active systemic drugs have been identified. Cisplatin has been extensively studied in both neoplasms and has clear activity (20% response rate in endometrial carcinoma and 23% response rate in squamous cell carcinoma of the cervix). Recently, paclitaxel (Taxol; Bristol-Myers Squibb Company, Princeton, NJ) has been shown to be clearly active in both (35% response rate in endometrial carcinoma and 17% response rate in squamous cell carcinoma of the cervix). The apparent clinical non-cross-resistance between paclitaxel and cisplatin in other neoplasms like ovarian carcinoma makes combinations including these two agents of great interest. In endometrial carcinoma, a phase I trial of cisplatin plus doxorubicin plus escalating paclitaxel doses is being performed by the Gynecologic Oncology Group (GOG). Based on the outcome of this study, a future randomized trial will compare the current standard, doxorubicin plus cisplatin, with either a combination of cisplatin, doxorubicin, and paclitaxel or a two-drug regimen of paclitaxel plus cisplatin. In squamous cell carcinoma of the cervix, a logical approach to the incorporation of paclitaxel into front-line therapy for advanced or recurrent disease is a phase III trial of the best regimen from GOG protocol 110 (cisplatin with or without either ifosfamide or dibromodulcitol) versus the same drugs plus paclitaxel. In addition, the GOG is conducting a phase I trial of paclitaxel given concomitantly with radiation in the hope that the resulting regimen will be an arm of a future randomized study in patients with locoregionally advanced disease (stages IIB through IVA). The ultimate role of paclitaxel in the management of patients with these two neoplasms awaits the results of these efforts. PMID- 7481865 TI - The treatment of early stage ovarian cancer. AB - Approximately one third of women with ovarian cancer present with localized disease. A series of recent studies have identified a population of patients who require only comprehensive surgical staging for optimal results and another group that may benefit from adjuvant therapy. A series of national and international studies are evaluating a variety of adjuvant treatments in prospective randomized trials that may enhance long-term survival in poor-prognosis early ovarian cancer. PMID- 7481866 TI - The role of paclitaxel in the treatment of head and neck cancer. AB - Current chemotherapeutic approaches to recurrent head and neck cancer have routinely yielded response rates of 10% to 30% (for single agents) and 30% to 50% (for combination chemotherapy). However, median survival times for patients with metastatic and/or recurrent disease have stagnated at around 6 months since the 1970s. The investigation of new drugs and treatment approaches thus continues to be a high priority. One such agent, paclitaxel (Taxol; Bristol-Myers Squibb Company, Princeton, NJ), has shown good single-agent activity and is also believed to potentiate the effects of radiation therapy in patients with head and neck cancer. We have focused on the addition of paclitaxel to the previously established combination of 5-fluorouracil, hydroxyurea, and radiation therapy. The study goals are to establish the maximum tolerated dose and dose-limiting toxicity of paclitaxel when added to this combination as a 5-day continuous infusion on a biweekly schedule. Preliminary results of this ongoing study have demonstrated activity in patients with poor-prognosis head and neck cancer. Major dose-limiting toxicities have consisted of neutropenia and mucositis, and definition of a recommended phase II dose is in progress. PMID- 7481867 TI - A clinical and pharmacokinetic study of high-dose carboplatin, paclitaxel, granulocyte colony-stimulating factor, and peripheral blood stem cells in patients with unresectable or metastatic cancer. AB - We have developed a regimen incorporating multiple cycles of high-dose carboplatin and fixed-dose paclitaxel (Taxol; Bristol-Myers Squibb Company, Princeton, NJ) with granulocyte colony-stimulating factor and peripheral blood stem cell support given every 21 days for up to four cycles. Our phase I study of this regimen has treated 26 patients with good performance status and histologically documented unresectable or metastatic carcinoma, sarcoma, or melanoma, 21 of whom received all planned courses every 21 days. Paclitaxel 250 mg/m2 was infused over 24 hours, followed by a 1-hour carboplatin infusion, with doses escalated between area under the concentration-time curve (AUC) targets of 8 and 20. Considering the carboplatin doses administered (two to three times those generally achieved with growth factor support), toxicity has been relatively modest. The median duration of grade 4 neutropenia and thrombocytopenia was not significantly different between the AUCs of 8 and 18, which proved to be the maximum tolerated carboplatin dose. Twelve courses were associated with hospitalization for neutropenic fever or catheter-related thrombophlebitis. One treatment-related death occurred, and severe toxicity caused withdrawal of two patients treated at the AUC of 20. Peripheral neuropathy was the most common serious nonhematologic complication. Pharmacokinetic analysis showed significantly lower measured versus predicted AUC values. Among 25 evaluable patients, preliminary results show one complete response (ovarian cancer) and 11 partial responses, including four in patients with non-small cell lung cancer. Additional issues to be addressed include the effect of a shorter (or longer) paclitaxel infusion on the carboplatin AUC (and the incidence of toxicity) and whether the discrepancy between actual and predicted AUCs (greater in our study than reported elsewhere) is due to the variability of creatinine clearance-determined glomerular filtration rate or to altered carboplatin pharmacokinetics when a short high-dose infusion follows paclitaxel. Additional patients are being accrued at the AUC of 18. PMID- 7481868 TI - Intraperitoneal paclitaxel in the management of ovarian cancer. AB - For several reasons, including its demonstrated activity in ovarian cancer, large size, hepatic metabolism, and lack of vesicant properties, paclitaxel (Taxol; Bristol-Myers Squibb Company, Princeton, NJ) is an interesting agent to consider for intraperitoneal administration in the management of ovarian cancer. Two phase I trials have confirmed a major pharmacokinetic advantage (> or = 1,000-fold) for peritoneal cavity exposure compared with the systemic compartment following intraperitoneal delivery of paclitaxel. A Gynecologic Oncology Group phase II trial of second-line intraperitoneal paclitaxel in ovarian cancer patients is currently in progress to determine whether the high concentrations and prolonged duration of exposure to paclitaxel of the surface of the peritoneal cavity can be translated into increased tumor cell kill and objective responses in this clinical setting. Future development of this novel therapeutic strategy will depend on the results of this important clinical trial. PMID- 7481869 TI - Carboplatin versus cisplatin in ovarian cancer. AB - The predominant data from clinical trials of advanced ovarian cancer have documented that carboplatin is equivalent to cisplatin in activity and causes considerably less ototoxicity, neurotoxicity, and nephrotoxicity. A large meta analysis of over 2,000 patients entered into phase III clinical studies showed that patients with advanced ovarian cancer had virtually identical survival durations when treated with carboplatin- versus cisplatin-containing regimens. Furthermore, in the recent National Institutes of Health Consensus Conference on Ovarian Cancer, it was concluded that "data from mature randomized clinical trials have indicated that the combination of carboplatin and cyclophosphamide is effective therapy" and that "the substitution of carboplatin for cisplatin leads to more acceptable toxicity." Cisplatin appears to be the analog of choice for intraperitoneal therapy, which has proven superior to intravenous (IV) therapy in a recently completed intergroup study. Thus, both analogs are likely to play an important role in therapy of curative intent for patients with stage III, optimal disease. Now that paclitaxel (Taxol; Bristol-Myers Squibb Company, Princeton, NJ) has proven to be an essential first-line agent in combination with cisplatin or carboplatin in the management of previously untreated, advanced disease, several new strategies have been developed. In the setting of phase III clinical trials by the Gynecologic Oncology Group (GOG) and the Southwest Oncology Group's (SWOG) Gynecologic Cancer Committee, for example, GOG-114/SWOG-9227 compares the standard IV paclitaxel/cisplatin regimen to a dose-intensive regimen incorporating two courses of carboplatin (area under the concentration-time curve = 9 mg/mL/min) plus intraperitoneal cisplatin/IV paclitaxel. The planned GOG replacement for this study will compare the standard IV paclitaxel (24-hour infusion)/cisplatin regimen to a paclitaxel (3-hour infusion)/carboplatin regimen. In the SWOG, two different high-dose regimens followed by autologous bone marrow transplantation are being evaluated in the setting of a phase II randomized trial. These regimens include high-dose carboplatin/cyclophosphamide/mitoxantrone and high-dose cisplatin/cyclophosphamide/thiotepa. The results of these and other GOG and SWOG trials will dictate the management of advanced ovarian cancer through the end of the century. PMID- 7481871 TI - [The protective effect of IL-1 beta on stress-induced gastric mucosal damage in rat]. AB - The effect of recombinant IL-1 beta on stress-induced gastric mucosal lessions was studied in rats. Pretreatment with IL-1 beta prevented formation of gastric mucosal damage in a dose-dependent manner. IL-1 receptor antagonist peptide (IRAP) could totally reverse the protective effect of IL-1 beta. IL-1 fragment peptide (163-171) had no effect on gastric ulcer formation in the experimental model, whereas sulfhydryl blocker N-ehtylmaleimide partially blocked the protective effect of IL-1 beta. The concentration of protein and noneprotein sulfhydryls in the gastric mucosa was significantly decreased 3 h after stress, and this decrease was partially prevented by IL-1 beta. IL-1 beta also decreased the concentration of molondialdehyde (MDA) in the gastric mucosa after 3 h stress. The results suggest that IL-1 beta could effectively lessen the degree of stress-induced gastric mucosal damage, due possibly to the production of endogenous sulfhydryl compounds in the gastric mucosa. PMID- 7481870 TI - Carboplatin in combination with paclitaxel in advanced ovarian cancer: dose determination and pharmacokinetic and pharmacodynamic interactions. AB - Data from various phase I/II studies of carboplatin in combination with paclitaxel (Taxol; Bristol-Myers Squibb Company, Princeton, NJ) have suggested that the degree of thrombocytopenia seen is less than that expected when carboplatin is given alone. However, some studies also have suggested that the area under the plasma concentration-time curve (AUC) of carboplatin is lower than that expected, raising the possibility of a pharmacokinetic interaction. Patients with advanced epithelial ovarian cancer were treated with first-line carboplatin (AUC = 7, using the 51Cr EDTA [edetic acid] clearance method) and escalating doses of paclitaxel. Thrombocytopenia was mild and was significantly less when the paclitaxel dose was 175 mg/m2 versus 150 mg/m2. Paclitaxel kinetics were nonlinear, as previously reported. The achieved carboplatin AUC was 7 +/- 1 mg/mL.min, indicating that the pharmacokinetics of carboplatin are not affected by paclitaxel. Glomerular filtration rates measured in 184 patients using the 51Cr EDTA clearance method were compared with rates estimated from the plasma creatinine level using the Cockcroft-Gault or Jeliffe formulas and showed a significant bias of these two formulas. Clearances above 50 mL/min were underestimated by an amount that became greater as the clearance increased and was approximately 25% to 35% for patients with clearances in the normal range. Since creatinine-based methods have been used in many previous studies, care is needed in interpreting the predicted AUC values from these studies. Carboplatin and paclitaxel may be given safely in combination at full doses, and the thrombocytopenia seen is significantly less than that observed with single-agent carboplatin. No evidence exists of a pharmacokinetic interaction, and the observation in some studies that the carboplatin AUC was lower than expected was probably due to the methodology used to estimate the glomerular filtration rate. PMID- 7481872 TI - [Role of locus coeruleus in analgesia caused by stimulation of supraoptic nucleus]. AB - The effects of stimulation of supraoptic nucleus (SON) on the changes of oxytocin (OT), arginine vasopressin (AVP), norepinephrine (NE) and serotonin (5-HT) in the perfusate of locus coeruleus (LC) and changes of pain threshild (PT) were studied by methods of microinjection, radioimmunoassay (RIA) and high pressure liquid chromatography (HPLC) technique. The results showed that the OT contents at 30, 60 and 90 min after stimulation, that of AVP at 30 min and 5-HT at 60 min were increased significantly, while the NE contents at 30 and 60 min were decreased markedly. Injection of V1 antagonist into LC did not affect the analgesic effect caused by administration of L-glutamic acid (L-Glu) into SON, which, however, could be partially inhibited by V2 antagonist or even reversed by OT antagonist. The above results suggested that the analgesic effect due to stimulation of SON is caused by increase of 5-HT and decreases of NE in the LC though the action of OT released by SON on OT and V2 receptor in LC. PMID- 7481873 TI - [Role of the lateroventral periolivary nucleus in the central chemoreception of respiration in rats]. AB - Using the methods of microinjection, microiontophoresis, electrolytic destruction and electric stimulation, the present work was undertaken to investigate whether the lateroventral periolivary nucleus (LVPO) might play a role in the central mechanism of chemoreception on respiration. Experiments were performed on 38 chloralose-urethane anesthetized Sprague-Dawley rats. The results were as follows: (1) Bilateral microinjection of acidified mock CSF into the LVPO region, evoked an enhancement in respiratory activities. (2) Microiontophoretic application of H+ caused mainly excitatory effects on the spontaneously discharging units in LVPO region, and some of the excitatory units were also responsible to microinjection of acidified mock CSF to the ventrolateral medullary surface. (3) After the electrolytic destruction of bilateral LVPO region in carotid body denervated rats, respiratory response to inhaled CO2 was attenuated significantly. (4) Electric stimulation of LVPO region caused an increase in discharging frequency of some units in the NTS and NRA regions. The above results indicate that LVPO plays a significant role in the central mechanism of chemoreception. PMID- 7481874 TI - [Impaired calcium uptake by cardiac sarcoplasmic reticulum and its underlying mechanism during rat septic shock]. AB - The underlying mechanism of Ca2+ uptake function of cardiac sarcoplasmic reticulum (SR) was investigated in the rat septic shock model produced by cecal ligation and puncture (CLP). The results are as follows. During the early phase of sepsis, the initial rate of ATP-dependent Ca2+ uptake by SR was decreased, while both the capacity of Ca2+ uptake and the activity of Ca(2+)-ATPase were unaffected. In the late sepsis, the impairment in SR function was even greater as the initial rate and the capacity of Ca2+ uptake by SR were significantly decreased, and this was paralleled by a reduction in Ca(2+)-ATPase activity. Although Ca2+ affinity (Km value) to calcium pump and the A0.5 values for Mg2+ and ATP activation on the Ca2+ uptake rate were unchanged, during sepsis the phosphorylation of SR vesicles by adding of catalytic subunit of the cAMP dependent protein kinase (PKA), calmodulin, or the fragment of PKC into Ca2+ uptake buffer, failed to stimulate Ca2+ uptake activities of SR isolated from early or late septic rats. These data suggest that depression of cardiac SR function is aggravated as sepsis develops, the impairment of SR Ca2+ uptake is possibly based on a mechanism of defective phosphorylation of SR rather than the ionic and energic regulatory actions of Ca2+, Mg2+, ATP on cardiac SR. PMID- 7481875 TI - [Effect of microinjection of sodium giutamate and glycine into ventrolateral nucleus of tractus solitarius on the evoked potentials of arcuatus nucleus in rabbits]. AB - Experiments were performed on 66 anaesthetized, vagotomized, paralyzed and artificially ventilated rabbits. The effects of microinjection sodium glutamate and glycine into ventrolateral nucleus of tractus solitarius (VLNTS) on hypothalamic arcuatus nucleus evoked potentials were observed. The main results obtained were as follows: (1) Phrenic nerve discharges were increased and the amplitude of P2 and N2 waves of arcuatus nucleus evoked potentials were decreased by microinjection of sodium glutamate into VLNTS. Phrenic nerve discharges were decreased and the amplitude of P2 and N2 waves of arcuatus nucleus evoked potentials were increased by microinjection of glycine into VLNTS. (2) After intravenous injection of naloxone, the phrenic nerve discharges could still be excited by microinjection of sodium glutamate as before, but the effects on the amplitude of P2 and N2 waves of arcuatus nucleus evoked potentials, unlike that of sodium glutamate, were reversed. The results suggest that the excitatory action of respiratory neurons in the VILTS may affect on the sensory function of arcuatus nucleus. The mechanism involved was discussed. PMID- 7481877 TI - [Alteration of phospholamban phosphatase activity associated with cardiac sarcoplasmic reticulum during sepsis in rats]. AB - In the present study, rat cardiac sarcoplasmic reticulum (SR) phospholamban (PLB) phosphatase was partially purified by chromatography on DEAE-Sephacel. This PLB phosphatase was indentical to phosphatase-1. It was shown on electrophoresis of SDS-PAGE autoradiography that the PLB phosphatase in rats during early sepsis (ES) depressed dephosphorylation of substrates (32P-phosphorylase a and 32P-SR). However, dephosphorylation of the substrates by the partially purified phosphatase during late sepsis (LS) was same as that in control rats. The partially purified PLB phosphatase activity in ES rats was significantly decreased, but showed no change in LS rats. The results above were confirmed by a studing of the substrate concentration (enzyme concentration, time)--enzyme reaction velocity curve in showing that both affinity and maximum initial velocity (Vmax) of the phosphatase in the ES rats were decreased, but had no change in those in the LS rats. PMID- 7481876 TI - [Altered ryanodine receptor of rat cardiac sarcoplasmic reticulum and its underlying mechanism during septic shock]. AB - The present study was undertaken to observe the changes of Ryanodine receptor of cardiac junctional sarcoplasmic reticulum (SR) in relation to membrane lipid microenvironment alteration during septic shock. The results showed that the Bmax for 3H-ryanodine binding to cardiac junctional SR was decreased by 41.3% (3.9 +/- 0.1 vs. sham 6.6 +/- 0.7 pmol/mg, P < 0.01) while the Kd value was unaffected during late septic shock (CLP 18 h). Ca2+ activated 3H-ryanodine binding significantly and reached a saturation value when Ca2+ concentration was 5 x 10( 5) mol/L, while the S0.5 and the Hill coefficient values remained unchanged during septic shock. Caffeine, ATP, and AMP-PCP activated while Mg2+, ruthenium red inhibited 3H-ryanodine binding in both groups but the A0.5 (concentration requires for half maximum activation) and the IC50 (concentration requires for half-maximum inhibition) for the above mentioned activators and inhibitors, were respectively unaffected during septic shock. Digestion of cardiac SR isolated from control rats with phospholipase A2 inhibited 3H-ryanodine binding, which could be dramatically recovered by the incorporation of phosphatidylcholine (PC), or phosphatidylserine (PS), or phosphatidylethanolamine (PE) into the isolated cardiac SR. Incorporation of above phospolipids into SR isolated from septic rats reversed shock-induced inhibition of 3H-ryanodine binding. It is concluded that the mechanism responsible for the inhibition of 3H-ryanodine binding of junctional SR during septic shock may be related to modification of membrane lipid microenvironment in response to PLA2 overactivation during septic shock. PMID- 7481878 TI - [Role of afferent renal nerves in 2K2C Goldblatt hypertension]. AB - In order to study the role of afferent renal nerves in 2K2C Goldblatt hypertension, the renal afferent nerves were selectively lesioned by bilateral T9 L2 spinal dorsal rhizotomy before clipping (internal diameter, 0.3 mm) of birenal arteries. Systolic blood pressure of the rat was measured by tail-cuff method. Concentrations of catecholamines, Ang II, and aldosterone were determined respectively by HPLC-EC and RIA, and vascular structural changes were measured by blood vessel micro-image analysis system coupled with a computer. The results showed that the concentrations of NE and E in medulla oblongata, adrenal gland and plasma, and of plasma Ang II and aldosterone as well as body weight of the rat were all significantly increased. The heart coefficient (heart wt/body wt), the media thickness, and the media thickness/lumen diameter in superior mesenteric arteries were also increased in 2K2C hypertensive rats (clipping, 6 wk) as compared with those in control rats. Bilateral rhizotomy delayed development of 2K2C hypertension and prevented above-mentioned vascular structural changes, the NE and E concentrations of medulla oblongata, adrenal gland and plasma were all decreased, hypothalamic NE and E were increased, and plasma Ang II level was not significantly changed. These results suggest that afferent renal nerves may play a partial role in the development of hypertension in 2K2C rats by activating sympathetic nervous system as a result of affecting metabolic activities of brain catecholaminergic neurons, and that high-plasma Ang II and aldosterone as well as heart hypertrophy and proliferation of vascular smooth muscle cells may also participate in the pathogenesis of hypertension in 2K2C rats. PMID- 7481879 TI - [Effect of angiotensin II on vasopressin gene transcription in the hypothalamus of rats]. AB - Experiments were carried out in male Sprague-Dawley rats. 3'-end digoxigenin labeled 26 bp oligonucliotide probe was used to detect the vasopressin (AVP) mRNA in the hypothalamus. Dot blotting technique was used in the investigation of AVP gene transcription level. The results showed that continuous intracerebroventricular administration (i.c.v.) of angiotensin II at a rate of 0.2 nmol/h for 2 days by using a miniosmotic pump resulted in a significant increase in daily water intake. An increase of AVP gene transcription level in the hypothalamus was also observed, but statistically insignificant. When daily water intake was limited (25 ml/d), continuous i.c.v. infusion of ANG II caused a significant increase in hypothalamic AVP gene transcription. It was also observed that hypothalamic AVP gene transcription level increased after salt loading and dehydration. However, intraperitoneal application of angiotensin converting enzyme inhibitor captopril (5 mg/(kg.d)) or i.c.v. nonpeptide angiotensin II receptor antagonist Dup753 (0.9 nmol/h) did not attenuate the increase of AVP gene transcription level induced by salt loading or dehydration. It is therefore suggested that the administration of ANG II enhances AVP gene transcription in the hypothalamus, especially when water intake is limited. However, this increase does not involve the participation of endogenous ANG II. PMID- 7481880 TI - [Effects of long-term atenolol treatment on beta-adrenoceptor subtypes in rat heart]. AB - The effects of long-term beta 1-AR selective antagonist atenolol treatment on beta-adrenoceptor subtypes were studied by radioligand binding assay, function determination and cAMP accumulation measerment in rat heart. The reasults showed that during long-term administration of atenolol: (1) The density of total beta AR was increased by approximately 57%; the positive inotropic response and cAMP formation induced by activation of beta-AR were also enhanced. (2) The 125I pindol competitive inhibition curve for CGP20712A showed that there were no significant difference in the percentage of beta 1- and beta 2-AR sites between the atenolol treated rats and the control rats; pA2 values for selective beta 1 AR antagonist CGP20712A and pKB values for selective beta 1-AR antagonist ICI 118, 551 were not significantly different in the two groups. The results suggested that beta 1- and beta 2-adrenoceptors were upregulated not only in density but also in positive inotropic effect to the same extent. PMID- 7481881 TI - [Excitatory effect of alpha-adrenoceptor on the transient inward current in sheep myocardium]. AB - Acetyl strophanthidin (AS) 0.05 mumol/L was used to induce a stable model of transient inward current (Iti) of sheep Purkinje fibers. The effect of the alpha adrenoceptor agonist phenylephrine (PE) 0.3, 1.0 mumol/L on the amplitude and duration of Iti was observed in the presence of propranolol 0.5 mumol/L to block the beta-adrenoceptor. When the preparation was perfused with PE 1.0 mumol/L for 20, 50 min, the amplitude of Iti decreased from control value 12.8 +/- 1.9 nA to respectively 10.7 +/- 1.2 nA (n = 5, P < 0.05) and 9.6 +/- 1.9 nA (n = 5, P < 0.01). The D50 of Iti prolonged correspondingly from control value 145 +/- 24.4 ms to 183.3 +/- 28.1 ms (n = 5, P < 0.05) and 207.5 +/- 34.2 ms (n = 5, P < 0.01). The inhibitory effect of PE on Iti showed a dose- and time-dependent manner. Both the time for arrival of the peak value and for return to baseline of Iti prolonged. This suggests that the kinetics of Iti channel changed in the presence of PE. When the Iti was enhanced by beta- adreceptor agonist isoprenaline (ISO) 1.0 mumol/L, the addition of PE 1.0 mumol/L to the perfusate for 10 min, the inhibitory effect become more pronounced, the amplitude of Iti decreased from 15.6 +/- 3.2 nA to 10.3 +/- 2.2 nA and the D50 of Iti prolonged from 92.5 +/- 14.3 ms to 132.5 +/- 36.0 ms (n = 5, P < 0.01). PMID- 7481882 TI - [Vasopressin facilitates aortic baroreceptor activity in the rat]. AB - Effect of arginine vasopressin (AVP) on the aortic baroreceptor activity of anesthetized rats was studied in 25 aorta arch-aortic nerve preparations. The results obtained were as follows: (1) By perfusing the isolated aortic arch with AVP (10(-8) mol/L), at constant intra-aortic pressure (IAP) of 13.3 kPa, the integrated aortic nerve activity (IANA) was increased by 40 +/- 4.7% (P < 0.0001), and the stepwise increase or decrease of IAP were accompanied correspondingly by changes of IANA. The function curve of aortic baroreceptor was shifted to the left and upward, and the peak into grated value (PIV) of the curve was increased from 621 +/- 85 to 711 +/- 81%.kPa-1 (P < 0.01). There were no changes in threshold pressure (TP), saturation pressure (SP), operating range (OR) and peak slope (PS). After washing out of AVP, the IANA returned to the control level. (2) The above effects of AVP were abolished by pretreatment with V1-antagonist [d (CH2)5Tyr- (Me) AVP, 10(-7) mol/L]. (3) By perfusing the aorta arch preparation with five different concentrations of AVP (10(-11)-10(-7) mol/L), at the same constant IAP, IANA were respectively 190%, 160%, 150%, 130% and 103% of the control. After pretreatment with V2-antagonist [d (CH2) (d Ile2Abu4] AVP (10(-7) mol/L), the perfusions with five different concentrations of AVP increased the IANA respectively to 130%, 150%, 160%, 170% and 180% of the control. When the V2-receptor-mediated vasodilating effect of AVP was blocked, the facilitatory effect of AVP on aortic baroreceptors via V1 receptors was significantly exhibited in a dose-dependent manner.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7481884 TI - [Observation on hypotensive action of adrenomedullin and its distribution in tissues]. AB - The effect of adrenomedullin (AM) on blood pressure and tension of aortic rings were studied in normal and in either spontaneous or renal hypertensive rats (SHR/RHR). The distribution characteristics of AM in viscera was also investigated. It was found that the hypotensive action of AM was far more prominent in hypertensive than in normotensive rats. The tension lowering action on the aortic rings is dose dependent and requires the presence of endothelium. Immunohistochemistry reactivity of AM of different degrees were also found in lung, kidney and myocardium, in addition to cells of pheochromocytoma and adrenomedulla. It was suggested that AM may play an important role in the regulation of blood pressure and visceral blood circulation. PMID- 7481883 TI - [Inhibitory effect and mechanism of scopolamine on respiration]. AB - In conscious rabbits, dogs and anaesthetized dogs, the respiratory frequency (FR), tidal volume (Vt) and minute ventilation (Vm) were determined in order to confirm whether the respiratory effect of scopolamine is stimulatory or inhibitory. Arterial blood samples were taken intermittently and analyzed for Po2, Pco2 and pH. When scopolamine was injected i.c.v., i.a. and i.v., the decrease of FR, Vt, Vm and Po2 and the increase of Pco2 were observed in two animals. When the drug was given in clinical doses used for treating respiratory failure, the effect was seen still observable. The results indicate that scopolamine inhibits respiration. Moreover, it was observed that excitatory effect of pilocarpine on respiration can be antagonized by scopolamine, but the inhibitory effect of 6 beta-acetoxy nortropane can not be antagonized, on the contrary, can be coordinated. The above results suggest that respiratory inhibition induced by scopolamine may be related to blocking M1-R of the respiratory center. PMID- 7481885 TI - [Induction of HSP70 mRNA in the brain and liver of rats after scalding]. AB - Induction of hsp70 mRNA in rats after scalding was analysed using 32P labeled hsp70 cDNA. The results showed that the expression of hsp70 mRNA in brain and liver was induced markedly after scalding. Time course of the increase showed that induction took place within 5 min and subsided 24 h after scalding. In the liver, induction took place still earlier, i.e. within 1 min and lasted for more than 24 h. The induction of hsp70 mRNA in rats after scalding did not seem to be due to elevation of core temprature since the increase of temperature of rat brain and liver after scalding was not more than 1 degree C. Western blot anlyses further showed that the induction of hsp70 occured within 5 min and lasted for more than 3 h in the brain of rats after scalding. The above results demonstrated that the hsp expression in internal organs could be induced by local injuries and was not necessarily dependent upon the direct effect of stimuli in intact animals. PMID- 7481886 TI - Hypercalciuria: is a low calcium diet appropriate? PMID- 7481887 TI - Simple urodynamic catheter placement. PMID- 7481888 TI - Getting ready for certification: voiding dysfunction. PMID- 7481889 TI - Can we talk with wives? PMID- 7481890 TI - Priapism: cause and treatment. PMID- 7481891 TI - Urinary incontinence in an acute care setting. AB - OBJECTIVE: To determine the prevalence of urinary incontinence in an acute care setting and the approaches to the management of it. STUDY DESIGN: An audit of hospital charts containing a diagnosis of urinary incontinence was conducted to gather data about patient demographics, related factors such as skin integrity and orientation, urinary tract infections, and nursing interventions. The audit was conducted by two data collectors who had an established interrater reliability at 100%. RESULTS: A 0.87% prevalence rate of cases with a diagnosis of urinary incontinence was found. An additional 13% of cases were documented only in nurses' notes. Nursing comfort measures were the primary choice the management of urinary incontinence. Bladder training and discharge planning were implemented infrequently. Additional findings regarding gender and age distribution, mental status, and management strategies are reported. CONCLUSIONS: Efforts to diagnosis and treat urinary incontinence must be directed to all age groups. Nursing protocols for accurate and consistent assessment, documentation, and discharge planning with follow-up should be implemented in the acute care setting so that patients can benefit from improved procedures to treat and manage urinary incontinence. PMID- 7481892 TI - Self-care strategies used for acute attack of interstitial cystitis. AB - OBJECTIVE: To determine the kinds of self-care used by women with interstitial cystitis and to find effective ways to manage symptoms of acute attack. STUDY DESIGN: One hundred thirty-eight women with interstitial cystitis completed a survey indicating how often they used and how effective they found strategies in four physical self-care subdomains (medication, treatment, diet, and body comfort) and three psychologic self-care subdomains (cognitive/stress reduction, distraction, and help-seeking). RESULTS: Descriptions of symptoms during an "acute attack" are more consistent with symptoms described "at onset" than symptoms experienced "currently." Physical strategies most used for acute attack included use of narcotics, antidepressants, and bladder analgesics, in addition to limiting the diet to bland or starchy foods, wearing loose clothing, taking hot baths, and using a heating pad over the pubis. Psychologic strategies most used included watching television, prayer, and seeking support from a significant other, family members, friends, and other patients with interstitial cystitis. CONCLUSIONS: Some psychologic and physical strategies found to be effective for other chronic pain conditions, such as biofeedback, alternating use of heat/cold, self-hypnosis, and redefining pain sensations were seldom used. Other nonmedical interventions such as massage and imagery, when used, were found to be more moderately to highly effective. Information about self-care and access to support groups may increase exposure to multiple ways of handling acute attacks. PMID- 7481893 TI - Electric stimulation and urinary incontinence: research and alternatives. PMID- 7481894 TI - Using condom seal to affix condom catheters. PMID- 7481896 TI - Dedifferentiated chondrosarcoma. AB - We reviewed 74 cases of dedifferentiated central and peripheral chondrosarcoma. Histologically these tumours consist of an underlying cartilaginous component (either benign or malignant) juxtaposed to a high-grade non-cartilaginous component, with a typically abrupt transition between the two tissue types. The non-cartilaginous component may constitute a very small or a very large proportion of the tumour, so diagnosis often requires histological evaluation of the entire tumour. The diagnosis is often suspected on the basis of the clinical course and careful evaluation of the radiographic characteristics. Central dedifferentiated chondrosarcoma can be classified radiographically into three types. In type I (36 cases in our review) the radiographic features are the same as those of a central chondrosarcoma, with the addition of a region with very aggressive radiographic features. Type 2 lesions (20 cases) resemble the underlying benign enchondroma but also have destructive changes and/or a large soft tissue mass. Type 3 lesions (8 cases) are not distinctive radiographically and present as a very high grade destructive lesion of bone. These cases are diagnosed following biopsy or tumour resection. The prognosis of these tumours is extremely poor, with 13% overall 5-year survival in this series. Improved survival was found in those cases where diagnosis was prompt and surgical treatment with a wide or radical margin was attained. No benefit was found from the use of adjuvant chemotherapy or radiotherapy. Thus, early recognition of the characteristic radiographic features, adequate histological sampling, and wide or radical surgical margins are necessary for satisfactory management of this highly malignant variant of chondrosarcoma. PMID- 7481895 TI - Juvenile spondyloarthropathies: clinical manifestations and medical imaging. AB - The spondyloarthropathies comprise four distinct entities--ankylosing spondylitis, psoriatic arthritis, the arthritis associated with inflammatory bowel disease, and Reiter's syndrome and other related forms of reactive arthritis. Although these are distinct diseases, they have a number of clinical, radiologic, and genetic characteristics in common which permit them to be classified under the unifying term "spondyloarthropathy". They are diseases of young adults, and when they present in patients under 16 years of age we refer to them as the "juvenile" spondyloarthropathies. They must be distinguished from juvenile rheumatoid arthritis, which is a totally separate entity; however the distinction may not always be obvious. Involvement of peripheral and sacroiliac joints commonly occurs in the juvenile spondyloarthropathies. The peripheral arthritis may be erosive and associated with bone apposition at the joint margins. Axial involvement is usually a late finding. Dactylitis and tenosynovitis are frequently present early on. Enthesitis, a highly specific feature, occurs much more often in the juvenile spondyloarthropathies than in the adult forms and it may be the only presenting feature. The plain radiograph is the primary and most important imaging modality for the assessment of these diseases. However, an expanding role of magnetic resonance imaging is evident. PMID- 7481897 TI - Determination of skeletal maturity by ultrasound: a preliminary report. AB - OBJECTIVE: Replacement of radiography by sonography for determination of skeletal maturity. DESIGN: Sonographic and radiographic evaluation of the maturation of the iliac bone apophysis (Risser's sign) and evaluation of the distal radial epiphyses. PATIENTS: 64 children and adolescents with idiopathic scoliosis, aged from 5 to 19 years. RESULTS: Sonograms with corresponding radiographs were available for the hand and wrist in 36 cases and for the iliac bone in 28 cases. Comparison of sonographic with radiological data revealed sonography to have a sensitivity for qualifying the status of the distal radial epiphysis of 93.1%. Specificity was 100%, accuracy 94.4% for investigation of the distal radial epiphysis. Overall accuracy for correct determination of Risser's stage by ultrasound (in comparison with the radiographic standard) was 89.2%. CONCLUSIONS: These findings suggest that ultrasound is a useful and highly acceptable tool for bone age determination in clinical routine or screening studies and will help to reduce ionizing radiation to children and adolescents. PMID- 7481898 TI - MRI appearance of meniscal cysts. AB - PURPOSE: The purpose of this study was to review the MRI criteria for the distinction of meniscal cysts from other cystic lesions in the knee so that appropriate treatment may be planned. DESIGN: A retrospective review of 62 knee MRI scans possibly showing meniscal cysts was performed. The type of meniscal tear, the appearance of the connection between the cyst and the meniscal tear, and the location, size, morphology, and signal characteristics of the meniscal cysts were recorded. Other types of fluid collections that had been mistaken for meniscal cysts were described. PATIENTS: Sixty-two patients were studied, ages 16 79 years, 61% male, 39% female. RESULTS AND CONCLUSIONS: Most of the meniscal cysts (91%) occurred immediately adjacent to the meniscal tear (98% horizontal cleavage tears, 49% anterolateral), with the tear leading directly into the cysts. In two cases, the cyst had dissected into the soft tissues distant from the meniscus and a connecting stalk was visualized. Fluid collections in normal bursae and recesses that had been mistaken for meniscal cysts had no direct connection to a meniscal tear. MRI can be used to distinguish meniscal cysts from other fluid collections that may mimic meniscal cysts. Pitfalls can be avoided by familiarity with the normal bursal and capsular anatomy, and by the application of specific diagnostic criteria. PMID- 7481899 TI - C1-2 arthrography. AB - OBJECTIVE: To describe the technique of C1-2 arthrography and recommend it as a suitable treatment for pain due to C1-2 abnormalities. MATERIALS AND METHODS: One hundred patients with the following conditions were studied: cervical pain or neuralgia without radiographic changes (group 1, n = 23), osteoarthritis (group 2, n = 37), rheumatoid arthritis (group 3, n = 23), ankylosing spondylarthritis (group 4, n = 5) and diverse conditions (group 5, n = 12). The technique consists of lateral puncture of the posterior aspect of the C1-2 joint with a 20-gauge needle under fluoroscopic control, arthrography using 1 ml contrast medium, and a 1-ml long-acting steroid injection subsequently. RESULTS: The articular cavity has an anterior and a posterior recess. Sometimes the posterior recess is large. In 18% of cases the contralateral joint also opacifies. CONCLUSIONS: C1-2 arthrography appears to be an efficient and safe technique for the treatment of upper cervical pain due to C1-2 articular disorders. PMID- 7481900 TI - Thickness of patellofemoral articular cartilage as measured on MR imaging: sequence comparison of accuracy, reproducibility, and interobserver variation. AB - OBJECTIVE: Since the thickness of cartilage is an important indicator of the status, progression and response to therapy of articular disorders, assessment of it is desirable. This study was undertaken to assess the accuracy, precision, and reliability of magnetic resonance (MR) measurements of articular cartilage. METHODS: Fifteen cadaveric patellas were imaged in the axial plane at 1.5 T. Gradient echo and fat-suppressed FSE, T2-weighted, proton density, and T1 weighted sequences were performed. We measured each 5-mm section separately at three standardized positions, giving a total of 900 measurements. These findings were correlated with independently performed measurements of the corresponding anatomic sections. A hundred random measurements were also evaluated for reproducibility and interobserver variation. RESULTS: Although all sequences were highly accurate (range r = 0.78-0.82), the T1-weighted images were the most accurate, with a mean difference of 0.25 mm and a correlation coefficient of 0.85. All sequences were also highly reproducible (mean difference between -0.09 and 0.05 mm) with little inter-observer variation (mean difference -0.04 and 0.11 mm). In an attempt to improve the accuracy of the MR measurements further, we retrospectively evaluated all measurements with discrepancies greater than 1 mm from the specimen. All these differences were attributable to focal defects causing exaggeration of the thickness on MR imaging. CONCLUSION: MR imaging is accurate, precise, and reliable as a basis for measuring articular cartilage and may potentially be usable to monitor progression of articular disorders. Care must be taken not to overestimate the thickness of areas with surface defects. PMID- 7481901 TI - Cat scratch disease: magnetic resonance imaging findings. AB - Cat scratch disease is an infectious lymphadenitis frequently occurring in children and adolescents. We present the magnetic resonance imaging findings of two patients with this disease. In both cases, lymphadenopathy was characterized by extensive stranding of the surrounding soft tissues, consistent with the inflammatory nature of this condition. Magnetic resonance imaging can be diagnostic and may obviate the need for invasive means of evaluation in patients suspected of having cat scratch disease. PMID- 7481902 TI - Grafted bone used in acetabular reconstruction: assessment by SPECT scintigraphy. AB - OBJECTIVE: To investigate incorporation of the grafted bone used for acetabular reconstruction. DESIGN: Single photon emission computed tomography (SPECT) was carried out at the 1st, 6th, and 12th month after the operation and the uptakes on serial SPECT scintigrams were normalized. The horizontal histogram of scintigraphic activity at the level of the central grafted bone in the coronal image was designated the profile curve. PATIENTS: Seven patients who had undergone bipolar hip arthroplasty for osteoarthritis of the hip were randomly selected. RESULTS AND CONCLUSION: Five of seven profile curves obtained at the 1st month demonstrated a single peak at the host-graft junction. The profile curve obtained at the 6th month showed two peaks at the host-graft junction and the periphery of the grafted bone in all patients. The profile curve at the 12th month showed slightly increased activity in the center of the grafted bone. This preliminary study demonstrates that comparison of the serial profile curves on SPECT is valuable in evaluating the progress of incorporation of the grafted bone in hip arthroplasty. PMID- 7481903 TI - Appendicular metastatic prostate cancer simulating osteosarcoma, Paget's disease, and Paget's sarcoma. AB - Skeletal metastases from prostate cancer is common and usually do not pose a diagnostic dilemma. This study reviews radiographic appearances of prostatic metastases to the appendicular skeleton in four patients where the appearances simulated osteosarcoma, Paget's disease and Paget's sarcoma. Prostatic metastases to long bones can produce appearances considered characteristic of other lesions and suggest misleading alternative diagnoses. PMID- 7481904 TI - Desmoplastic fibroma of the calcaneus. PMID- 7481905 TI - Osteoma of the innominate bone with intraosseous and parosteal involvement. AB - A case has been presented of a 47-year-old woman with a dense bony mass both within and on the surface of her right public ramus. This was discovered incidentally on plain radiographs of the pelvis. Besides osteoma, a differential diagnosis of parosteal osteosarcoma, ossifying parosteal lipoma, periostitis ossificans, osteochondroma with attenuated cartilage cap, and melorheostosis was considered. Histological evaluation of open biopsy samples showed typical findings of an osteoma. This is an unusual case of an osteoma with both parosteal and intraosseous involvement. PMID- 7481906 TI - Intraforaminal schwannoma of the sacrum. PMID- 7481907 TI - Small cell osteosarcoma of the soft tissue. AB - The lesion we described is an osteoid producing mitotically active spindle cell soft tissue neoplasm, which has light and ultrastructural features of an osteosarcoma. By immunohistochemistry it marks with p30/32MIC2-directed antibodies but fails to react with antibodies associated with other soft tissue neoplasms. The precise histogenesis of the lesion is unknown, but failure to elicit a periosteal reaction makes the periosteum less likely to be the primary tissue of origin. The failure to demonstrate any radiologic or histologic evidence of involvement of the underlying bone leaves the soft tissue as the most reasonable site of origin, although the parosteal region cannot be completely excluded. PMID- 7481908 TI - Low-grade chondrosarcoma in a 12-year-old boy. AB - We present a case of a rare central chondrosarcoma in a 12-year-old boy and discuss its radiographic and magnetic resonance appearance. PMID- 7481909 TI - Impacted fracture of the femoral head secondary to recurrent posterior hip dislocation in a child. PMID- 7481910 TI - Shoulder girdle lipomatosis. AB - Shoulder girdle lipomatosis is a distinct type of lipomatosis which has unique clinical and radiographic features. The MR findings reflect the pathologic findings and, in our opinion, allow the differentiation of shoulder girdle lipomatosis from other lipomatous masses on the basis of imaging findings alone. PMID- 7481911 TI - Piriformis bursitis causing sciatic neuropathy. PMID- 7481912 TI - Chondrosarcoma in a patient with relapsing polychondritis. AB - A case of a chondrosarcoma in a patient with relapsing polychondritis was presented. The association of these two unusual disorders of cartilaginous tissue has never been described before. The clinical presentation and radiologic and histologic features were discussed. PMID- 7481913 TI - Demographic and socioeconomic links to cigarette smoking. AB - This paper illuminates the demographic and socioeconomic factors associated with smoking statuses. It employs the 1990 National Health Interview Survey's Health Promotion and Disease Prevention Supplement and logistic regression to examine the covariates of smoking status among the U.S. adult population. Polychotomous logistic regression, which provides an alternate way to examine smoking practices, simultaneously considers multiple variables and their interactions, controls for other important covariates, produces predicted values and patterns, and allows multiple comparisons. By examining interactions, we have found that age displays distinct, often curvilinear, patterns with smoking; that compared to females, males have higher rates of cigarette consumption except at the youngest ages; that Anglos, especially Anglo males, exhibit high probabilities of cigarette consumption but also high probabilities of being former smokers; that Black males exhibit high probabilities of light smoking, but only at the older ages, and that they also exhibit high probabilities of being former light smokers; and that Mexican-American females are the least likely to currently smoke or to have ever smoked. By clearly elucidating the demographic and socioeconomic variations in smoking, arguably the most salient marker of health behavior, we can better target programs and policies geared toward the reduction of smoking-related diseases and deaths. PMID- 7481914 TI - Effects of lactation and contraceptive use on birth-spacing in Bolivia. AB - Based on the 1989 Demographic and Health Survey of Bolivia, analysis of the joint effects of breastfeeding and contraceptive use on birth-spacing showed the IUD to be the most effective contraceptive method used to delay conception. Breastfeeding significantly lengthened the birth interval, but only following second and higher parity births. In addition, conditions of poverty appeared to further inhibit the return of fecundity and delay conception. PMID- 7481915 TI - Gender and performance in athletics. AB - A comparison of women's and men's performance in swimming, running, and skating is made based on available world record data. The ratio of women's to men's record times is analyzed as a time series, and the asymptotic value of this ratio is discussed in the context of a model. "Rate of improvement curves" are defined over time and by event in a given sport and are compared across gender. The relationship of record time to event distance is estimated and also analyzed across event and gender boundaries. We make three conclusions: 1. The rate of improvement for women has been extraordinary and is larger for longer distance vents. 2. Law-like relations emerge for record times and event lengths. These relations have interesting physical interpretations when examined for various sports. 3. The male physiology is more suited to anaerobic strength events while, given increased access and participation, women can be expected to be more on a par with men in some long-distance aerobic events. PMID- 7481916 TI - Another look at mortality crossovers. AB - Considerable debate still ensues on whether or not the so-called "mortality crossover," the intersection of age curves of mortality at the older ages, is artifactual or real. One school of thought argues that it is a function of misreporting of ages, greater for one population than another. The other school of thought counters that it persists apart from age misreporting and is due to selective frailty processes as age increases. An historical review of the debate is reported, after which an attempt at a balanced conclusion is offered. PMID- 7481917 TI - Quantitative approaches to detect the fourth stage of the epidemiologic transition. AB - Widely diverse quantitative measures have been used to study the onset of the different stages of the epidemiologic transition, including the fourth stage. This report suggests additional methods for detecting the fourth stage of the epidemiologic transition that are more parsimonious than those previously used. While these methods are not new, they have potential for application in this area of research. An example of one of these methods is illustrated, using Canadian data. PMID- 7481918 TI - Effect of maternal age on birth outcomes among young adolescents. AB - This study examined the effect of maternal age on birth outcomes among young adolescents, ages 10 through 15. All records representing single births of primipara, Black or White adolescents, were selected for analysis from the 1983 1986 National Center for Health Statistics' Public Use Linked Live Birth-Infant Death Data File (n = 127,668). Logistic regression analyses controlled for effects of maternal race, marital status, prenatal care, gravidity, education, and metropolitan/nonmetropolitan residency. Univariate analyses indicated that the youngest adolescents were at greatest risk for negative birth outcomes including very preterm and preterm delivery, low birth weight, small for gestational age (SGA), and neonatal mortality. Logistic analyses showed similar results, with the exception that differences in SGA were insignificant. This study indicates the importance of examining age-specific birth outcomes among a population that has traditionally been studied in aggregate and underscores the need for increased prevention efforts. PMID- 7481919 TI - Emergence of diabetes mellitus in a Mexican-origin population: a multiple cause of-death analysis. AB - The "New World syndrome" is comprised of disorders that are hypothesized to have resulted from an interaction of the Amerindian genotype with an environment that includes marked changes in lifestyle and diet. The principal component of the syndrome is adult-onset (noninsulin dependent) diabetes mellitus. The purpose of this paper is to describe the emergence of diabetes in a Mexican-origin population. Using a unique file of multiple-cause mortality data, we have computed standardized mortality ratios and relative standardized mortality ratios for Mexican-origin individuals and for other white persons age 30 and over from the 1930's through the middle 1980's. Results for the study population residing in Bexar County (San Antonio), Texas, show that diabetes mortality for Mexican origin individuals did indeed increase in a pattern consistent with the New World syndrome hypothesis. This study is the first description of the emergence of diabetes using a data set with consistently defined causes of death and demographic characteristics. PMID- 7481920 TI - Effects of gender, birth order, and other correlates on childhood mortality in China. AB - Using data from the 1988 Two-Per-Thousand Survey of Fertility and Birth Control, this paper examines the effects of gender, birth order, and other correlates of childhood mortality in China. Controlling for family-level factors, childhood mortality is found to be associated with the child's gender and birth order. Among firstborn children the difference between male and female childhood mortality is not statistically significant, but among others, female children between ages 1 and 5 experience higher mortality than male children. Childhood mortality is slightly higher for children who have older brothers only than for those who have older sisters only, and it is highest for those who have both older brothers and sisters. Other factors affecting childhood mortality in China include mortality of older siblings, birth interval, urban/rural residence, mother's level of education, and mother's occupation. All interactive effects between gender and family-level characteristics are found to be statistically insignificant. PMID- 7481921 TI - The marriage squeeze and the rise in informal marriage in Brazil. AB - Around the world, populations have experienced shortages of one sex or the other at marriageable ages, as a result of mortality declines. The solutions to this problem vary with the cultural context. Declines in the spousal age difference and increases in dowry payments (India) and polygamy (Africa) are two adjustments to a disequilibrium in the marriage market. We hypothesize that in Brazil the marriage market finds its balance by "recycling" men through highly unstable informal unions. Using census and 1984 survey data, we establish the relationship between a marriage squeeze and the increase in informal marriage. Census data and a competing-risks analysis of marriage choice provide evidence that a marriage squeeze has affected both the chances of marrying at all and the type of marriage entered. PMID- 7481923 TI - Parental attachment, drug use, and facultative sexual strategies. AB - It is often asserted that sexual promiscuity and drug abuse appears to share a common etiology in poor parental attachment. This study explores this claim empirically among 480 college students. Other variables--religiosity, masculinity/femininity, sex, age, and physical appearance--that may enhance or reduce the incidence of promiscuity and drug use were included in multivariate analyses. Parental attachment was significantly related to both variables, and the combination of poor parental attachment and drug use was a strong predictor of promiscuity for both males and females. In multivariate analyses, religiosity was the most important predictor of promiscuity for males, and attachment was the most important for females. The findings are examined guided by the three desiderata commonly accepted as relevant to biosocial attachment theory. PMID- 7481924 TI - Understanding the AIDS epidemic and reacting sensibly to it. PMID- 7481922 TI - Birthweight-specific infant mortality risks for Native Americans and whites, United States, 1960 and 1984. AB - We used NCHS natality and linked-birth/death certificate tapes to compare birthweight-specific neonatal and postneonatal mortality risks for Native Americans and whites in 1960 and in 1984. The birthweight distributions for the two groups were similar both years. Native American neonatal mortality risk dropped from 20.2 in 1960 to 5.2 in 1984, and the relative risk for Native Americans with respect to whites fell from 1.31 in 1960 to a nonsignificant difference in 1984. Postneonatal mortality risks for Native Americans fell from 27.5 in 1960 to 6.2 in 1984, with a drop in the relative risk from 5.2 to 2.1. Although the relative improvements for Native Americans were highest in postneonatal survival, Native Americans still had over twice the level of white postneonatal mortality. Birthweight was positively associated with survival for both groups, but the odds of Native American neonatal death were affected less by low and very low birthweights. For both groups, improvements in neonatal mortality were highest at the lower birthweights, while the gains in postneonatal survival benefitted normal and high birthweight infants most. PMID- 7481925 TI - Stigma, HIV and AIDS: an exploration and elaboration of a stigma trajectory. AB - Stigma is a social construction which dramatically affects the life experiences of the individuals infected with the human immunodeficiency virus (HIV) and their partners, family and friends. While it has been generally recognized that the nature of stigma varies across illnesses, it has usually not been considered as changing and emerging over the course of a single illness. In this paper, HIV/AIDS is analyzed in terms of a stigma trajectory. The primary purpose is to conceptualize how individuals with HIV/AIDS experience stigma and to demonstrate how these experiences are affected by changes in the biophysical dimensions of HIV/AIDS. Four phases of the HIV/AIDS stigma trajectory are depicted: (1) at risk: pre-stigma and the worried well; (2) diagnosis: confronting an altered identity; (3) latent: living between illness and health; and (4) manifest: passage to social and physical death. The essential processes through which individuals personalize the illness, dilemmas encountered in interpersonal relations, strategies that are used to avoid or minimize HIV-related stigma, and subcultural networks and ideologies that are drawn upon to construct, avow, and adapt to an HIV identity are considered across the stigma trajectory. PMID- 7481926 TI - Exploring the iceberg of morbidity: a comparison of different survey methods for assessing the occurrence of everyday illness. AB - This paper examines problems in measuring the occurrence of acute symptoms of ill health. Health interview surveys and health diaries often lead to different results. Two key hypotheses assume that: 1, interviews using checklists are more sensitive to the respondent's psychological distress than are the open-ended questions of health diaries; and 2, health diaries demand high levels of compliance leading to underreporting of symptoms. An additional 3rd hypothesis assumes that the effect of psychological distress on response patterns is strong for reporting psychological symptoms but insignificant for musculoskeletal symptoms. The hypotheses were tested and explored with data from the Dutch Survey of General Practice, a nationwide study among 161 GPs. A random sample of 100 patients per GP was approached for a health interview and asked to keep a structured health diary during three weeks. Symptoms were recorded during the interview with a checklist and queried in the health diary with open-ended questions. The occurrence of symptoms was modelled with logistic regression. High levels of psychological distress increase the likelihood of recording symptoms for both instruments, but the increase is greater for the interviews. Respondents who have only received limited education, heavy smokers and those who suffer from chronic conditions have a significantly lower likelihood of recording symptoms in the diary as compared to the questionnaire. There was no significant effect of taking an interest in health matters, gender, and work and domestic role obligations. Taking the nature of symptoms into account, it was found that psychological distress had indeed a great effect on the response pattern for psychological symptoms, but not for musculoskeletal symptoms. The criticism that symptom checklists are sensitive to psychological distress rather than to physical illness alone, is confirmed in this study. Open-ended questions prevent biased responses, but result in fewer symptoms being recorded. Health diaries with open-ended questions 'produce' more symptoms but take more effort to complete, requiring sufficiently motivated respondents. It is recommended that a less biased specific list for the assessment of acute symptoms be developed. PMID- 7481927 TI - U.S. apartheid and the spread of AIDS to the suburbs: a multi-city analysis of the political economy of spatial epidemic threshold. AB - We compare mechanisms of AIDS diffusion at the county level from five U.S. central city epicenters into their associated metropolitan regions. Four of the five show an expanding 'hollowed out' center of physically and socially devastated, politically and economically abandoned high density minority neighborhoods, surrounded by rings of relatively affluent majority suburban populations. From these centers AIDS diffuses into the suburbs as a single, spatially extended disease ecosystem. The exception, San Francisco, has not yet experienced the 'hollowing out' process and is, we conclude, a major AIDS epicenter markedly less coupled to its suburbs because of that fact. This may constitute one of the few empirical observations of spatial threshold in epidemiology. Our empirical results contradict the conclusions of a recent National Research Council report that AIDS will be largely confined within marginalized urban populations. In reality U.S. urban apartheid, particularly its continuing disruption of minority social structures, has markedly accelerated the diffusion of AIDS into suburban communities. A widespread program of reform, which rebuilds minority physical and social community structures within both city and suburb, is an essential, but largely unrecognized, component to any serious strategy for the control of AIDS in the United States. PMID- 7481928 TI - Decisional conflict in general practice: strategies of patient management. AB - Exploratory interviews were carried out with 37 general practitioners as an initial stage of a larger project investigating medical decision making in primary care. Qualitative analysis of free responses to a set of semi-structured questions revealed a common set of variables which influence decisions about patient management. These included: time pressure, uncertainty and patient characteristics. Results corroborate previously reported findings and may have important implications for the variation of management decisions about similar patient complaints. Strategies of patient management are identified, and the correspondence between these and the postulates of decisional conflict theory explored. PMID- 7481929 TI - Physician participation in Medicaid managed care. AB - Medicaid programs throughout the United States are moving away from fee-for service medicine and increasingly towards managed care models in attempts to control rising health care costs. This study examines the participation of physicians in Arizona's prepaid, managed care Medicaid program, the Arizona Health Care Cost Containment System (AHCCCS). In particular, it considers the reasons that physicians decide to participate in AHCCCs, and for participants, the possible impact of these reasons on whether they participate fully or on a limited basis. A mail survey was sent to 300 primary care physicians in Arizona, of which 171 completed surveys were returned. Results suggested that physicians tend to participate because of reimbursement, approval of the 'type' of medicine that managed care allows them to practice, and their belief in Medicaid-type programs. Physician attitudes toward the patients, while generally negative, do not keep them from participating in Medicaid. However, the type of patients in AHCCCS is related to physician satisfaction with the AHCCCS health plans, as is reimbursement. The study indicates differences in the reasons for participation between Arizona's physicians and those in fee-for-service Medicaid programs of other states. PMID- 7481930 TI - Determinants of persistent compliance with screening for colorectal cancer. AB - Mass population screening for colorectal cancer is currently being evaluated by means of randomized controlled trials. These trials point to the likelihood that, if implemented, the level of both initial and sustained compliance will prevent the full potential of screening being realised. The paper opens by reviewing the evidence on determinants of compliance, both initial and longer term, although little empirical evidence on adherence to repeated screening is currently available. The paper then presents the results of a survey of persistent compliers and non-compliers within the English screening trial, in order to identify those characteristics most closely associated with persistent compliance behaviour. Persistent compliers are found, inter alia, to be of higher socio economic classes than persistent non-compliers, to have more personal and family experiences of illness and to visit their dentists more regularly. The results suggest that generalized attempts at compliance enhancement would be ineffectual against the prevailing background characteristics of the non-compliant population, and that the more overt targeting of efforts in this respect is to be preferred. PMID- 7481931 TI - Nemawashi essential for conducting research in Japan. AB - Nemawashi is a semi-formal but systematic and sequential consensus building procedure in Japan by which the approval of a proposed idea or project is sought from every person in a significant organizational position. For foreigners planning research in Japan, this concept has important implications since the project approval process is more obscure than in many Western countries. In this paper, I discuss observations as an outsider the research environment and culture in a Japanese hospital as seen from the inside, and draw conclusions for conducting research in Japan. From May to July of 1992, I was supported by the Japan-United States Educational Commission (the Fulbright Program) to interview physicians about end of life decision-making in Japan. The proposed project sought to obtain information on the way Japanese physicians use family and patient preferences in clinical decision-making. As I initiated the project, my Japanese advisor took great pains to explain the need for nemawashi to gain approval in my host institution. He underscored the importance of carefully informing every clinical and administrative person who might be affected by the proposed project, as well as the steps necessary for obtaining their endorsement. This process alone took three weeks and personal negotiations by my advisor at six levels before final project approval was granted by the dean. For actual data collection, my advisor made personal introductions to a high ranking physician from each department which greatly facilitated the scheduling of subsequent interviews with other faculty members in that department. The personal introductions by my host professor ensured 100% participant cooperation.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7481932 TI - Potties, pits and pipes: explaining hygiene behaviour in Burkina Faso. AB - Stool disposal practices have been shown to be associated with childhood diarrhoea. However, efforts to promote improved hygiene behaviour are hampered by a lack of understanding of what determines those behaviours. Data from 2793 household interviews with mothers of children from the town of Bobo-Dioulasso in Burkina Faso were analyzed to examine what differentiated mothers who reported using safer stool disposal practices from those who did not. Three 'outcomes' were considered: where the child was reported to defaecate; where the mother reported disposing of the child stools; and whether excreta were observed in the compound. Regression models were developed to identify those factors with the strongest independent associations with the outcomes. There was a consistent association between the source of water and the outcomes. Mothers with access to a tap in the yard reported using safe hygiene practices three times more often than mothers using wells outside the compound and twice as often as mothers who used public standpipes or wells within the yard. The source of water showed a similar pattern of association with observations of faecal matter in the environment. Improved sources of water may contribute to safer stool hygiene by reducing the time spent on water collection or by encouraging mothers to conform to higher standards of hygiene. Other factors which played a role in predicting the hygiene behaviour of mothers were the husbands' occupation, the number of health education sessions that she had attended, her zone of residence and family ownership of certain valuable objects. These factors are likely to be related and to be, to some extent, proxies for the real determinants of her behaviour. A model of the cultural, psycho-social and infrastructural proximate determinants of hygiene behaviour is proposed. Data from focus group discussions suggested that the main purpose of hygienic behaviour is to conform to existing norms of social etiquette. Trials of interventions based on changing such norms are needed to test whether this is an effective means of promoting of safer hygiene practices. PMID- 7481933 TI - Structural relationships between social support and coping. AB - Relationships between social support and coping were examined over a one-year period in a sample (n = 120) exposed to a specific stressor (i.e. a spinal cord injury). Two issues were evaluated: (1) patterns of social support and coping over time; and (2) the direction of the effects of coping on social support or vice versa. Subjects had incurred a spinal cord injury within the preceding year, completed their rehabilitation, and returned to the community. They were assessed at 1, 4 and 12 months post-discharge to capture possible changes in patterns of social support and coping. Covariance structure modelling indicated a single factor construct based on three measured subscales for social support (instrumental/informational/emotional), and for coping (problem oriented/perception-oriented/emotion-oriented). While the factor structure of coping was invariant over time, the structure of social support changed from a high concentration of informational support at one month to a higher saturation of emotional support at 4 and 12 months, potentially reflecting changes in the salience of different components of the network. The perceived availability of social support was seen to have direct effects on future coping. At one month, social support had a direct positive effect on coping at four months. However, at four months social support had a negative effect on coping at 12 months, which may reflect a change in the structure of the social support construct itself. That is, as the factor structure of social support varied with time, coping may also have been influenced by differing perception of one's needs and circumstances. Thus, the study provides evidence for the dynamic effects of social support on coping, depending on one's stage in the process of long-term adjustment. The findings underscore the interplay between social support and coping, and the need for future research and practical applications to recognize temporal effects on the relationships between these two complex constructs. PMID- 7481934 TI - AIDS knowledge and risk behaviors among domestic clients of female sex workers in Bali, Indonesia. AB - This study investigated AIDS and STD knowledge, risk behaviors and condom use among clients of female commercial sex workers in Bali, Indonesia. Although these clients were varied in their socioeconomic status, they all tended to have low levels of knowledge concerning HIV and STD transmission and prevention, multiple sexual partners, low frequencies of condom use with these partners, and experienced frequent STDs. Although HIV sero-prevalence rates are currently low in Indonesia, clients of CSWs are at high risk of HIV transmission. Given their high mobility and frequent sexual encounters with CSWs in other parts of Indonesia, these men could well be agents of rapid spread of the virus throughout Indonesia. Interventions to prevent the spread of the HIV virus must be targeted not only to CSWs, but also to their clients. These interventions should include educational activities concerning AIDS and STD transmission and prevention, condom promotion, efforts to improve condom availability, and activities to strengthen the health sectors' STD diagnosis and treatment capabilities for both CSWs and their clients. PMID- 7481935 TI - Socio-economic differences in smoking: Dutch adolescents' beliefs and behaviour. AB - Smoking behaviour is more frequent among those who have a low than those who have a high socio-economic status. The present study confirms that this is also true for Dutch adolescents. The purpose of this study was to investigate whether adolescents with a low socio-economic status (LSES) have different motives for smoking or non-smoking than those with a high socio-economic status (HSES), the aim being to formulate more specific recommendations for prevention strategies for the various groups. HSES adolescents perceived a stronger association between smoking and a number of health-related, personal and social disadvantages, while linking smoking more clearly with the discovery of the taste and with relief from boredom than LSES adolescents. LSES adolescents viewed smoking as a way of meeting people. HSES adolescents experienced less positive norms and less social pressure towards smoking than did LSES adolescents. Smoking among LSES adolescents seems to be embedded in the social culture, and has a stronger social function. Smoking prevention programmes for this group will have to be integrated into a broader, community-based approach, which should also pay attention to the smoking behaviour of the social environment in general and that of the parents in particular. Finally, improved legislation with regard to smoking prevention will be required, in order to facilitate the adoption and implementation of smoking prevention programmes. PMID- 7481936 TI - Socioeconomic equity in Finnish hospital care in relation to need. AB - The aim of the study was to evaluate the success of Finnish health care policy in establishing socioeconomic equity in the use of hospital inpatient care. We studied the use of short-term (< 30 days) care at Finnish general hospitals among those aged 25 or over, psychiatric and obstetric patients excluded. The data on service utilization were obtained from the 1988 Finnish Hospital Discharge Register. Patient data were linked with socioeconomic indicators from the 1970 1987 population censuses by personal identification number. The data on population at risk were obtained from the 1987 census. Hospital utilization was measured by annual risk of hospitalization, discharge rate, and inpatient days. The socioeconomic distribution of hospital utilization according to need was assessed by mortality and morbidity data. The same data were used to calculate inequity indices. Low socioeconomic groups used more hospital services than high in all age-groups and both genders. The socioeconomic differences in hospital utilization were similar to the gradients in death rate or to the prevalences of poor self-perceived health and limiting long-standing illness. In relation to need, the lower socioeconomic groups used at least as much inpatient care as the higher. The inequity index showed a neutral distribution of hospital services with respect to need. Finnish health care policy in the late 1980s seems to have been successful in providing hospital care equitably. This study compared overall hospital use with overall mortality and morbidity. It did not address possible socioeconomic differences in hospital use by causes of hospitalization or the quality of hospital services provided. PMID- 7481937 TI - Acceptability of prenatal diagnosis of sickle cell anaemia (SCA) by female patients and parents of SCA patients in Nigeria. AB - The acceptability of prenatal diagnosis (PND) of sickle cell anaemia (SCA) as a means of controlling sickle cell disorder in Nigeria was examined using a structured questionnaire. The respondents were comprised of 92 adult female patients with SCA, aged 15-20 (23 +/- 6) years; 53 HbAS mothers, aged 20-61 (37 +/- 11) years and 48 HbAS fathers, aged 33-65 (46 +/- 8) years. More than 85% of the respondents would like PND to be offered in Nigeria and 92% of the HbAS mothers as well as 86% of the fathers would like to have the investigation or allow their wives to go through the procedure, respectively. Only 35% of the patients as compared to 63% of the mothers and 51% of the fathers would opt for termination of an affected pregnancy. Fear of the complications of abortion and religious convictions were the two most frequently cited reasons for opposing pregnancy termination. The high percentage of parents who would opt for termination of an affected pregnancy was associated with the societal emphasis on perfection and on previous experience in management of SCA patients. It is interesting that most of the respondents still favoured effective genetic counselling as the best means of controlling SCA. PMID- 7481938 TI - Smoking and absence from work: Australian evidence. AB - This study reports on research into the relationship between absence from work and smoking. A key feature of the study is the data, which come from the National Health Survey (NHS) undertaken in 1989/90 in Australia. Involving responses from more than 54,000 individuals, the NHS provides what is almost certainly the largest and most comprehensive data set available in the world today containing information on both absence and smoking behaviour. Moreover, the data permit controls to be applied for a large number of influences thought to have some bearing on work attendance. Logit models of absence incidence over a two week period are estimated, and smoking is consistently found to have a large and significant impact on absence. This impact, however, is not consistent across the sexes. The probability of a male smoker being absent from work is estimated to be 66% greater than that for a male who has never smoked. For females, the corresponding figure is just 23%. The findings also suggest that it is important to distinguish ex-smokers from other non-smokers, with the incidence of absence among ex-smokers being almost as high as that for current smokers. Finally, no evidence was uncovered to suggest that absence varied with the actual quantity of tobacco smoked, as measured by both the number of cigarettes smoked and estimated daily nicotine and tar intakes. PMID- 7481939 TI - Health care reform in the United States: on the road to nowhere (again)? PMID- 7481940 TI - Cultural studies of biomedicine: an agenda for research. AB - This paper outlines a 'cultural studies' approach to investigations of the transnational world of contemporary biomedicine. Although biomedicine is fostered by an international political economy and global community of medical educators and bioscientists, it is taught, practiced, organized and consumed in local contexts. This essay argues that cultural studies of contemporary biomedicine should focus on the dynamic relationship between local and international worlds of knowledge, technology and practice. Three issues illustrate this approach: (1) an exploration of the tensions inherent in the local and cosmopolitan shaping of 'clinical narratives', with examples drawn from comparative studies of oncology; (2) an exploration of the influence of biomedical research findings and international clinical trials on the production of clinical narratives, with examples drawn from current research on breast cancer; and (3) an exploration of the local or national and 'international' or 'transnational' dimensions of the production of biotechnologies and pharmaceutical therapeutics. The essay concludes with a discussion of the limits that privilege either universal or local perspectives and claims to knowledge and the ethical challenges that become apparent from this perspective. PMID- 7481941 TI - Equity of health care in Australia. AB - This paper examines the equity characteristics of health care financing and delivery in Australia and compares its performance with recent findings on systems in Europe and the United States. Vertical equity of finance is evaluated with income and payment concentration indices derived from published survey data on taxes and expenditure by income decile. Horizontal equity of health care delivery is assessed with standardized expenditure concentration coefficients for three measures of health status and four types of health services, derived from household survey data on health care utilization, health status, income and demographics. Health cover is available to the entire population. Results show the financing system is slightly progressive despite the fact that 30% of payment comes from private sources, which are regressive. The equity index compares favorably to many European countries and is much better than the U.S. which has a regressive financing system. The Australian system fares less well in terms of equity of health care delivery. Several features favor privately insured higher income persons in use of health care and this is reflected, for some health status measures and types of service, in inequity favoring the better off. This contrasts with inequity favoring the less well off in many European countries and the U.S. This analysis provides a benchmark for monitoring the equity of the Australian system and provides information on the equity of a mixed private and public financing system that covers the entire population. This is relevant to the U.S. which is moving in this direction by extending private cover to the uninsured and to European countries that are increasing private sector involvement in health care financing. PMID- 7481942 TI - The relationship between cost-effectiveness analysis and cost-benefit analysis. AB - This paper examines the relationship between cost-effectiveness analysis and cost benefit analysis. Provided that a cost-effectiveness analysis includes all the relevant societal costs, it is shown that a cost-effectiveness analysis can be interpreted as a cost-benefit analysis where the willingness to pay per effectiveness unit is assumed to be constant and the same for everyone. To relax this assumption the willingness to pay per effectiveness unit can be allowed to vary depending on for instance the size of the health effects and the target population. It is argued that cost-effectiveness analysis is best viewed as a subset of cost-benefit analysis, where the aim of the analysis is to estimate the cost function of producing health effects. It is also concluded that to interpret and use cost-effectiveness analysis as a tool to maximize the health effects for one specified real-world budget, will be inconsistent with a societal perspective and is likely to lead to major problems of suboptimization. PMID- 7481943 TI - The social and political meaning of the silent epidemic of miners' phthisis, Bendigo 1860-1960. AB - Miners' Phthisis was suffered by goldminers in the Central Victorian city of Bendigo, Australia for 100 years from the 1860s to the 1960s. This paper examines the disease using the framework put forward by Karl Figlio in a study of miners' nystagmus and later adapted by Evan Willis to consider the social processes of Repetition Strain Injury. It demonstrates that a study of industrial disease does not require moral judgements on the disease's biological reality or medical controversy about its treatment in order to fit within such a framework. More, it argues that without an understanding of the social and political meaning of the disease historically and progressively, the biological has itself little meaning. PMID- 7481944 TI - Notions and treatment of guinea worm in northern Ghana. AB - Dracunculiasis, infection with Dracunculus medinensis or guinea worm, is widespread in the Northern Region of Ghana, where rural people drink from unprotected water sources such as ponds and small-scale dams. This paper discusses the results of an anthropological study of beliefs and practices concerning commonly occurring illnesses, such as infection with guinea worm (nierifu), in two rural Dagomba communities in the Northern Region of Ghana. The importance of knowing about local perceptions and treatment of guinea worm is stressed. Guinea worm is not attributed to water. The general understanding is that guinea worm is an innate part of human anatomy. It is not seen as an alien presence in the body. Guinea worm is rather said to be 'in people's blood', and sooner or later to 'stand up'. Guinea worm is considered an 'inevitable' feature of living. After a description of the background to the study, the methods are characterized. Brief background information on the people, their environment and their water sources are given. The central portion of the paper focuses on local perceptions of illness and notions of guinea worm ('guinea worm is in the human blood'), which are very different from those of biomedicine ('guinea worm is a disease'). Attention is also given to perceptions of water ('bitter' vs 'sweet') and the prevention of guinea worm. The social limitations to the filter technology are addressed. People's choice of therapy and the role of medicines (herbs and Western pharmaceuticals) in treatment of guinea worm are also considered. The paper concludes with a discussion of health education and stresses the importance of showing respect for the local view of guinea worm, which is said to be 'in the blood'. It is suggested that, since people are not adverse to the use of Western pharmaceuticals, the use of Western medicines to treat guinea worm should be further promoted. The social constraints on filtering must also be appreciated. These relate to the organization of labour within the household and the dominant role of the 'senior wife'. PMID- 7481945 TI - General practitioners' assessment of and interest in alternative medicine in Canada. AB - Canadian physicians' opinions about alternative medicine have, as yet, not been assessed. The objectives of this pilot study were to assess general practitioners': (1) desired involvement in alternative medicine; (2) perceived demand for alternative medicine; and (3) beliefs about the efficacy of different alternative approaches. The study design was a cross-sectional survey of 400 randomly selected Alberta and Ontario general practitioners. Of the 384 eligible physicians, 200 (52%) completed the questionnaire. Seventy-three percent of physicians felt that they should have some knowledge about the most important alternative treatments. However, with respect to other issues, physicians desired less involvement with alternative medicine. Sixty-five percent perceived a demand for alternative medicine from their patients, in particular chiropractic. Alternative medicine was perceived to be needed most for musculoskeletal problems and chronic pain or illness. Chiropractic, hypnosis and acupuncture (for chronic pain) were believed to be most efficacious, while homeopathy and reflexology were considered to be least efficacious. Undergraduate, graduate clinical and continuing medical education will need to address alternative treatments in order to provide physicians with up-to-date and relevant information. PMID- 7481946 TI - Powerlessness, health and mortality: a longitudinal study of older men and mature women. AB - National samples of older men (age 45-59 in 1966) and mature women (30-44 in 1967), surveyed periodically over more than a decade, establish the association over time between the sense of powerlessness and various indices of health status (chiefly, psychosocial symptoms and limits on physical activities). The results are basically coordinate for men and women, and they are replicated for initially healthy and initially impaired sub-samples. The results show that: (1) in each year, powerlessness is associated with greater activity limits and more psychosocial symptoms; (2) powerlessness also provides prospective prediction, since high initial powerlessness scores are associated with health problems observed five and ten years later, with initial health controlled; (3) increasing powerlessness accompanies deterioration in health (with stringent controls on prior health); and (4) for a sub-sample of men, mortality between 1976 and 1981 is also associated with initially high powerlessness scores (with prior health controlled). These results are discussed for their import in relation to the steadily growing interest in social psychological factors in health. PMID- 7481947 TI - Maternal activity budgets: feasibility of exclusive breastfeeding for six months among urban women in Honduras. AB - Although exclusive breastfeeding for 6 months is likely to be beneficial for infant health, mothers often cite time pressures as a reason to introduce other foods. Using 12-hr in-home observations at 19 and 24 wk postpartum, we compared maternal activity budgets and time spent breastfeeding among low-income, urban mothers not employed outside the home who exclusively breastfed (EBF) or gave other foods in addition to breast milk (BF+SF). Breastfeeding time was similar between groups (averaging approx 75 min/12 hr) except that multiparous EBF mothers at 24 wk spent more time breastfeeding their infants than did BF+SF mothers. However, total time devoted to infant feeding (including preparation and feeding of solids in the BF+SF group) was significantly greater in the BF+SF group than in the EBF group (except among multiparous women at 24 wk). Feeding frequency per 12-hr averaged 6.7-8.9 in the EBF group and 5.4-8.1 BF + 1.8 meals in the BF+SF group. Both EBF and BF+SF mothers had considerable 'free' time; they spent, on average, 2-3 hr during each 12-hr period resting, chatting or watching TV. Primiparous women tended to spend less time at household chores and child care than multiparous women. Of the BF+SF mothers, 60% said that breastfeeding was 'somewhat' or 'very' time-demanding (vs 15% of EBF mothers), 49% said that it interfered with other activities (vs 6% of EBF mothers), and 26% said that they gave other foods to extend the time between breastfeeds. Although many of the BF+SF mothers knew that giving foods may increase infant illness, there were cultural pressures not to exclusively breastfeed. For example, many introduced solids to their infants prior to 6 months because they believed that infants should 'learn' how to eat other foods, to 'acclimate' the stomach and learn to differentiate flavors. We conclude that time constraints are not a barrier to exclusive breastfeeding in this population, although they may be perceived as a barrier. PMID- 7481948 TI - Activists and delegators: elderly patients' preferences about control at the end of life. AB - The views of elderly patients concerning control over the dying process are explored in this paper. Thirty-eight hospitalized elderly patients were interviewed about their attitudes, both general and personal, toward exerting control over end-of-life decisions. A majority (27) of the participants could be categorized as 'activists', that is, they preferred to have a voice in decision making at the end of life. There were also a number of 'delegators' (11) in the study. They preferred to delegate the decision-making to their physicians, to God, or to fate. Activists were better educated, had held more professional and managerial jobs and tended more often to be middle class rather than lower class. Their advanced sophistication about consumer rights and their general knowledge of health issues is reflected in their greater interest in patient control over dying. Most activists rejected the idea of euthanasia or assisted suicide for themselves, but favoured the withholding and withdrawing of treatment. It is possible that these ideal types of 'activists' and 'delegators' could anchor a continuum that is measurable. Such a scale could facilitate the differentiation of the elderly on this issue and thereby prove useful in the development of appropriate policy. PMID- 7481949 TI - Predictors of emergency room use by homeless adults in New York City: the influence of predisposing, enabling and need factors. AB - Employing data from a 1987 shelter survey of 1260 homeless adults in New York City, multivariate models of emergency room (ER) use are developed which include an array of risk factors for visiting a hospital ER including health and mental health problems, victimization and injuries. The study's primary goal is to identify factors that predict ER use in this population. Multivariate logistic and linear regression models were tested separately for men and women predicting three outcomes: any use of the ER during the past 6 months, use of the ER for injuries vs all other reasons (given any ER use), and the number of ER visits (given any ER use). Lower alcohol dependence, health symptoms and injuries were strong predictors for both men and women; other significant predictors differed markedly by gender. Both models were highly significant and produced strikingly high risk profiles. A high prevalence of victimization and injuries underlies ER use among the homeless. Based upon the findings, we recommend expanded health and victim services as well as preventive measures. Until primary care becomes available for this population, we advise against policies that discourage ER use by the homeless. PMID- 7481950 TI - Attitudes towards people with HIV: are they as stigmatizing as people with HIV perceive them to be? AB - Stigma is a feature of HIV disease and many people who are HIV-positive report that their lives are affected by fear of discrimination (felt stigma). Although opinions do not necessarily predict behaviour, this article examines whether the public's avowed attitudes to people with HIV are as punitive and stigmatizing as those infected think they are, and the extent to which public attitudes may contribute to felt stigma. A street survey was conducted in Glasgow and Edinburgh, Scotland, asking a stratified quota sample of 300 men and women of all ages to complete a short questionnaire about their attitudes toward people with HIV. The same questionnaire was also completed by 42 men and women with HIV. One in five respondents in the street survey, and all of those with HIV, were also asked to complete the questionnaire imagining that they were a typical member of the public, to find out whether both groups attributed more hostile attitudes to generalized others than they themselves professed. Overall, the general public had relatively liberal views about people with HIV although a majority felt that some restrictions should be placed upon their freedom. Controlling for age, sex, socioeconomic status and city, people with HIV had more liberal attitudes than the general public, but perceived public attitudes to be far less liberal than were reported in the street survey. Respondents in the street survey also perceived generalized others to be less liberal than themselves. These results provide evidence of felt stigma among people with HIV and the policy implications are discussed. The findings are also set within a theoretical framework concerning the nature of attitudes, their relationship to behaviours, and the pervasiveness of negative images associated with AIDS. PMID- 7481951 TI - Family planning practices and attitudes among former Soviet new immigrant women in Israel. AB - One hundred young new immigrant women from the former U.S.S.R. now living in Israel answered a detailed semi-open questionnaire regarding their knowledge, attitudes and behaviors in fertility and birth control issues. A collective family planning profile of these women is largely in line with that of the urban population of Slavonic U.S.S.R., combining early marriage, early and low fertility, the latter achieved by both abortion and contraception. Most respondents and their partners tried to prevent unwanted pregnancies, usually starting from traditional methods and switching over time to modern ones. An IUD remained most popular contraceptive among parous women, while use of the pill, very rare in the U.S.S.R., has almost doubled upon migration, mostly among younger women. Still, they kept some misleading ideas on the pros and cons of traditional versus modern methods, suggesting lack of adequate information also upon migration. Like their ex-compatriots, our women preferred to solve their birth control problems without external professional involvement. Contrary to the expected, free abortion ideology was not universally advocated by our respondents, and most were fully aware of abortion limitations in Israel. While rationally condemning abortion in both moral and health terms, most respondents still use it, this gap between beliefs and practice being indicative of their readiness to adopt efficient contraception. This switch occurs faster in women actively involved with host society via work or studies. Younger women were found to be more flexible and advanced in their family planning practices than were older ones, while almost no differences were related to education and origin within the U.S.S.R. This exploratory study suggests that any investment into promotion of healthy fertility control practices among new immigrants will be cost-effective in the near future. PMID- 7481952 TI - An assessment of research on breastfeeding promotion strategies in developing countries. AB - Breastfeeding and its impact on child survival in developing countries have been well documented. Governments are being urged to encourage breastfeeding through legislation and promotional campaigns. The success of promotional programs depends not only on the interventions themselves but on the acceptance and acquisition of the desired knowledge, skills and behaviors. During the past decade, a variety of strategies have been used in an attempt to promote breastfeeding. These efforts include: (1) modifying hospital policies; (2) using social supports; (3) providing incentives; (4) educating mothers and health workers; and (5) initiating legislation and political action to create policies aimed toward healthier infant feeding practices. Research regarding these promotional activities has been both qualitative and quantitative in nature, designed to provide answers concerning the relative success of different promotional approaches. To varying degrees, findings and conclusions of breastfeeding promotion research can enhance the design, implementation and sustainability of these projects. However, programs and research designs are poorly described in the literature, providing vague conclusions and little guidance for future program planning. PMID- 7481953 TI - Strategy of risk approach in antenatal care: evaluation of the referral compliance. PMID- 7481954 TI - Pulmonary sarcoidosis: population differences and pathophysiology. AB - Sarcoidosis is characterized as a hyperimmune response to an unknown agent(s) at the lesion sites. Regarding pulmonary lesions, macrophage-T lymphocyte alveolitis precedes an epithelioid cell granuloma formation. In the 35 years since the first International Conference on Sarcoidosis, the definition of sarcoidosis has been revised several times, and many new pathophysiologic mechanisms have been proposed. In this review, we focused on four important issues: (1) possible population differences, (2) possible causative agent(s) and the current problems of disease susceptibility or resistance, (3) recent data on pathogenesis (including a genetic approach) and pathophysiologic processes, and (4) monitoring of disease activity. PMID- 7481955 TI - Medical characteristics of 61 unwarranted malpractice claims. AB - We describe malpractice claims closed by plaintiff attorneys after initial medical expert (ME) review. This retrospective study includes 61 cases, of which 33 originated in hospitals, 18 in physicians' offices, and 10 in nursing homes. No claims were against state or federal agencies. The reviews were done between 1987 and 1994 for 29 attorneys from 11 states. The cases fell into 12 diagnostic groups, and leading causes for action were late diagnosis (13 cases), drug administration (11 cases), sepsis (6 cases), and strokes (5 cases). We conclude that internists are particularly liable to unwarranted malpractice claims when unexpected complications occur and when treatment fails to cure infectious disease. However, such claims may be terminated following the initial ME review. PMID- 7481956 TI - Office or emergency department: what's the difference? AB - Many studies have examined why patients with nonurgent conditions seek care in emergency rooms. These studies have evaluated financial concerns, demographic variables, and proximity of the emergency room. However, none of these studies have attempted to predict which patients with nonurgent conditions will seek care at the emergency room and which will postpone medical treatment until regular office appointments can be obtained. Demographic and psychological variables help predict which patient will go to the emergency room for treatment. Since the demographics cannot be changed, health professionals must focus on problem solving skills to lessen the burden on our already overcrowded emergency rooms. PMID- 7481957 TI - Tuberculosis screening in adults who have received bacille Calmette-Guerin vaccine. AB - The incidence of tuberculosis among immigrants to the United States is 12 times that in the native-born population. Screening immigrants for tuberculosis is complicated by the widespread use of bacille Calmette-Guerin (BCG) vaccination. To determine the utility of tuberculin testing in adults who have been vaccinated with BCG vaccine, we studied the tuberculin reactions of 80 adults who came to us for naturalization physical examinations. No adverse effects were reported from tuberculin testing. Subjects from regions with a low prevalence of tuberculosis who had received BCG vaccine were significantly more likely to have a positive reaction than subjects who had not received BCG vaccine. However, among subjects from regions with a high prevalence of tuberculosis, there was no difference in the prevalence of positive reactions between those who reported having BCG vaccinations and those who said they had not. Interpretation of tuberculin reactions in immigrants who receive BCG vaccinations depends on the prevalence of tuberculosis in the country of origin. Adults receiving BCG vaccination who have a positive reaction and no evidence of active tuberculosis should receive prophylactic therapy or be observed carefully. PMID- 7481958 TI - Predicting fetal lung maturity by visual assessment of amniotic fluid turbidity: comparison with fluorescence polarization assay. AB - We prospectively studied 159 patients having clinically indicated amniocentesis. Amniotic fluid (3 to 5 mL) was placed in a nonheparinized glass tube. This sample was then classified as turbid (indicating maturity) or clear (indicating immaturity) on the basis of a single examiner's ability to read newspaper print through the glass tube. These results were then compared with fluorescence polarization values for the same sample. A value of 70 mg/g was considered positive evidence of fetal lung maturity. By study criteria, 62 samples (39%) indicated immaturity and 97 (61%) indicated maturity. Turbidity correctly identified 89 samples that produced fluorescence polarization values of at least 70 mg/g. Turbidity as a predictor of fetal lung maturity when compared with fluorescence polarization assay has a 91% positive and 87% negative predictive value. Visual inspection of amniotic fluid may be of value in areas where sophisticated methods are unavailable. PMID- 7481959 TI - Discussions with outpatients with chronic obstructive pulmonary disease regarding mechanical ventilation as life-sustaining therapy. AB - Patients with acute exacerbations of chronic obstructive pulmonary disease (COPD) are often too sick to make informed decisions regarding the use of mechanical ventilation (MV). Prior knowledge of their wishes regarding this form of therapy, therefore, is essential in order to preserve their autonomy. In order to characterize this group of patients with respect to their knowledge of mechanical ventilation as a potential treatment for their disease, we surveyed 40 stable outpatients with the diagnosis of COPD. All patients attended either the general medical clinic or the pulmonary clinic of a university and veterans' hospital. The median age of the patients was 67 years, and the median forced expiratory volume in 1 second was 1.26 L. Consecutive patients were asked to participate in a survey that examined the following questions: (1) Was there an awareness of MV as a treatment modality? (2) Had there been prior discussions with their physicians concerning MV? (3) Were the patients able to express a decision concerning MV as indefinite life-sustaining therapy? Our results showed that 35% of patients were not aware of MV. Only 14% had previously discussed MV with their physicians; 40% indicated a decision to use MV, 22% would decline MV, and 38% were unsure. Patients who had discussions were more likely to specify a choice concerning MV when compared with those patients who had no prior discussions. We conclude that discussions between patients with COPD and their physicians occur infrequently, but when they do occur, they help patients formulate a decision. PMID- 7481960 TI - Hyponatremia in pediatric patients with HIV-1 infection. AB - Hyponatremia has been recognized as a complication in adults with acquired immunodeficiency syndrome (AIDS). We did a retrospective study evaluating the medical records of 86 children (age 4 months to 21 years) with human immunodeficiency virus (HIV-1) infection to determine the frequency and clinical associations of hyponatremia. Twenty-two children (26%) developed hyponatremia (serum sodium < 135 mEq/L; range 104 to 134 mEq/L; mean 130 mEq/L). Fourteen were male; 18 of the 22 patients were black and 4 were white. At the time of hyponatremia, the children frequently had comorbid associations, including 8 (35%) with AIDS encephalopathy; 3 (14%) with cardiomyopathy; 3 (14%) using diuretics; 1 (5%) using pentamidine; 3 (14%) with bacterial pneumonia; 2 (9%) requiring gastric lavage feedings; 2 (9%) with tuberculosis meningitis; 2 (9%) with gastroenteritis; 1 (5%) with infection caused by Mycobacterium avium intracellulare; 1 (5%) each with brain tumor and tumor metastasis to brain. The cause of hyponatremia was attributed to syndrome of inappropriate antidiuretic hormone in 8 children; poor sodium intake and/or excessive diarrheal losses in 5; and the use of diuretics in 3 patients. Mild hyponatremia with no identifiable cause was found in 5 patients. PMID- 7481962 TI - Ozone level effect on respiratory illness: an investigation of emergency department visits. AB - This project examined the relationship of daily ozone (O3) levels to the number of respiratory-related emergency department (ED) visits. The confounding effects of temperature, humidity, mold, and pollen counts were examined, as were the effects of day of the week on numbers of ED visits. The design is a time-series relating concurrently measured variables that are repeatedly measured over time. Respiratory-related ED visits at three hospitals in a southern city were examined. Visits were analyzed separately for pediatric, adult, and geriatric patients. A significant association was found between elevated O3 levels and respiratory-related visits for adults. The same association was not found for pediatric or geriatric ED visits. Days with higher ozone were accompanied by higher numbers of adults going to the emergency department because of respiratory complaints. However, the effect was not large and does not appear to hold for pediatric or geriatric ED visits. PMID- 7481963 TI - Factors associated with participation in a preventive cardiology service by patients with coronary heart disease. AB - A free preventive cardiology service for patients and their asymptomatic family members was established at a large teaching hospital in the southeastern United States to provide medical triage and risk reduction counseling for prevention of coronary heart disease. Patients and families were informed about the service and invited to attend during a personal visit from members of the preventive cardiology consultation service. Our study was designed to explore determinants of decisions to attend the clinic. We conducted telephone interviews with 62 consecutive patients to collect data on individual perceptions of health risks, factors that modify perceptions of health risks and ability to make behavior changes, and actions taken to reduce risk of disease. Of the 62 patients interviewed, 39% had attended the clinic. Patients hospitalized for myocardial infarction or bypass surgery were more likely to attend the clinic than those who had angioplasty or catheterization without bypass surgery. PMID- 7481961 TI - Prospective survey of continuous versus single-injection spinal anesthesia in obstetrics. AB - Prior to the removal of spinal microcatheters from the market in 1992, these catheters were used extensively in the Obstetric Anesthesia Service at the Medical Center of Louisiana. We report on a prospective survey of the clinical application of single-injection versus continuous-catheter spinal anesthesia. Two hundred sixteen patients had single-injection anesthesia, and 218 had continuous catheter anesthesia. No neurologic complications other than postdural puncture headache (PDPH) were encountered in either group. Five patients had PDPH after single-injection technique, and 8 patients had PDPH from continuous spinal anesthesia. Patients with diabetes were at higher risk for PDPH, and blacks were at lower risk (relative risks 4.35 and 0.31, respectively). Additionally, PDPH was associated with lower intraoperative urine output. No increased risk of complications was found after continuous spinal anesthesia with microcatheters. PMID- 7481964 TI - Percutaneous dilatational tracheostomy: an alternative approach to surgical tracheostomy. AB - Percutaneous dilatational tracheostomy (PDT) offers a nonoperative alternative to patients needing an elective tracheostomy. The technique associated with PDT is essentially a progressive dilatation using blunt-tipped dilators for the placement of a tracheostomy tube. We retrospectively compared patient charges and operating time for 18 PDTs and 18 surgical tracheostomies. Our analysis suggests that percutaneous dilatational tracheostomy is a cost-effective alternative to the open procedure for elective tracheostomy. PMID- 7481965 TI - Rhabdomyolysis and myoglobinuric acute renal failure associated with classic heat stroke. AB - Classic heat stroke is a disorder of thermal regulation that predominantly affects elderly patients during heat waves. In contrast to exertional heat stroke, rhabdomyolysis and myoglobinuric acute renal failure are considered to be unusual manifestations of classic heat stroke. We retrospectively reviewed the charts of seven patients admitted to Maimonides Medical Center with classic heat stroke over a 3-day period during a heat wave in July 1993. Three of these patients with classic heat stroke had rhabdomyolysis, but no renal failure; two completely recovered; and one had an ataxic gait disturbance. Three additional patients had rhabdomyolysis and myoglobinuric acute renal failure; one of them completely recovered, one survived with quadriplegia, and one died. Our findings suggest that rhabdomyolysis and myoglobinuric acute renal failure are common manifestations of classic heat stroke. Recognition of this complication warrants rigorous hydration and alkalinization of the urine to prevent or attenuate myoglobinuric acute renal failure. PMID- 7481966 TI - Clonazepam-sensitive intermittent dystonic tremor. AB - We report three cases and a literature review describing a syndrome of intermittent idiopathic focal or segmental dystonic tremor dramatically responsive to clonazepam. All patients were young men who had intermittent symptoms. After magnetic resonance imaging of the brain and laboratory analysis of blood, each patient was treated with clonazepam and clinically observed for 1 year. In all three cases clonazepam produced full abatement of the tremor. None of the patients displayed progression of symptoms, and all have remained tremor free with stable doses of clonazepam. Clonazepam-sensitive intermittent dystonic tremor may represent a benign syndrome occurring in young men. PMID- 7481967 TI - Cardiopulmonary complications during laparoscopy: two case reports. AB - Laparoscopic surgery is growing in popularity. As a result, laparoscopic procedures are being done on a broader and older patient population. These patients may have underlying cardiopulmonary disease that predisposes them to complications not seen in younger patients. Anesthesiologists should be aware of this possibility and of the problems inherent to the pneumoperitoneum necessary for laparoscopy. We present two cases involving elderly patients to illustrate cardiopulmonary complications that can occur during establishment or maintenance of the increased intra-abdominal pressures required for laparoscopic surgery. The first case describes a patient who developed bradycardia and asystole during insufflation for a laparoscopic hernia repair. The second case involves severe hypercarbia and a pneumothorax due to massive subcutaneous emphysema that developed during a laparoscopic colon resection. PMID- 7481968 TI - Pulmonary hyalinizing granuloma: an unusual association with multiple sclerosis. AB - Pulmonary hyalinizing granuloma (PHG) is a rare entity included in the differential diagnosis of pulmonary nodules of unknown origin. The pathologic entity may represent a peculiar form of pulmonary immune reaction. We report the case of a 40-year-old white woman who had nodular lesions in both lung bases after a 10-year history of multiple sclerosis. An open lung biopsy was required to make the diagnosis. The association of pulmonary hyalinizing granuloma with multiple sclerosis could be coincidental, but since there is a possibility of immune reaction in the pathogenesis of both diseases, the association may be significant. PMID- 7481969 TI - Intramural abscess of the duodenum resulting from perforated peptic ulcer. AB - Even in this age of effective medical treatment for peptic ulcers, their complications remain the same. Many require prompt surgical intervention. We present an unusual case of postbulbar duodenal ulcer complicated by intramural perforation and abscess formation. Such complications have a high mortality rate because of frequent delays in diagnosis and treatment caused by a lack of peritoneal signs or other specific clinical findings. PMID- 7481970 TI - Crohn's disease masquerading as systemic lupus erythematosus. AB - We report the case of a 55-year-old woman who was thought to have systemic lupus erythematosus (SLE) for more than 13 years before Crohn's colitis was correctly diagnosed. The differentiation of Crohn's disease from SLE may be difficult, as illustrated by our case. Gastrointestinal symptoms, laboratory studies, and radiographic findings may appear similar between the two diseases. The correct diagnosis has important treatment and prognostic implications. PMID- 7481972 TI - Hormone therapy for gastrointestinal bleeding in hereditary hemorrhagic telangiectasia. PMID- 7481971 TI - Snake-strike--induced ischemic colitis with colonic stricture complicated by disseminated intravascular coagulation. AB - A 57-year-old farmer was struck on the right thumb by a pit viper (Agkistrodon blomhoffu). Subsequently, he had acute renal failure and disseminated intravascular coagulation (DIC), associated with melanotic stools and abdominal pain. Renal failure caused by renal cortical necrosis was successfully treated with hemodialysis. A double-contrast barium enema examination revealed multiple stenoses of the colon, regional edema, and longitudinal ulcer. Histologic examination of the stenotic lesions after laparotomy revealed fibrosis of both submucosa and proper muscle layer, with fibrotic thickness in the small arteries of the colonic wall, indicating that ischemic colitis was associated with DIC. In this case, DIC from viper toxins played an etiologic role in the development of ischemic colitis with stricture, as well as acute renal failure. PMID- 7481973 TI - Psychiatry in medicine. PMID- 7481974 TI - Acute severe autonomic insufficiency during pentamidine therapy. PMID- 7481975 TI - Chronic fatigue syndrome (myalgic encephalopathy). AB - Chronic fatigue syndrome is associated with many misconceptions. In this review, we attempt to summarize various pathogenic hypotheses for this disease and discuss new lines of insight into causes and treatments of this baffling and most frustrating condition. PMID- 7481976 TI - Herpes zoster and internal malignancy. AB - Herpes zoster (HZ) often occurs concomitantly with various internal malignancies, most commonly hematologic in origin. Authors in the past proposed that HZ was a marker for internal malignancy, since it is often found in association with a malignancy. A critical review of the literature revealed that when HZ and malignancy occur in the same individual, rarely does HZ precede the malignancy, but usually follows it. Many studies have evaluated HZ in cancer patients, but only three studies have evaluated the occurrence of malignancy after the diagnosis of HZ, and all found no increased incidence of internal malignancy in patients with HZ. Since HZ is a poor marker for internal malignancy, extensive workups to find occult malignancy are not indicated. In rare cases, however, HZ precedes a malignancy. We therefore recommend a baseline history and physical examination, with further directed workup only if there are abnormal findings. PMID- 7481977 TI - Glove safety: summary of recent findings and recommendations from health care regulators. AB - Every health care worker should be aware of the quality and deficiencies of gloves in the health care setting. Since the introduction of the human immunodeficiency virus into the health care setting, the medical community's compliance with universal precautions and the standards mandated by the Food and Drug Administration have worked in a mutual effort to enhance the quality of medical gloves considerably. Despite these improvements in protecting health care workers, recent studies on glove integrity before and after use warn against taking the protective aspect of gloves for granted. According to studies, latex gloves provide the best available protection from biohazardous substances, but stricter regulation of glove production and a continual observance of recommendations and regulations by health care regulators are necessary to improve glove quality. This report is designed to reveal the fact regarding the true safety integrity of two main glove types, latex and polyvinyl chloride, through permeability testing with various biomaterials and substances. PMID- 7481978 TI - Is tenure irrelevant for academic clinicians? AB - Academic tenure is the guarantee that a faculty member cannot be arbitrarily dismissed. The tenure system developed to maintain academics free of external pressure and coercion, and in dramatic instances tenure has been used to defend academic freedom. With the development of the research-oriented medical school staffed by full-time faculty, the reward system of tenure and promotion was grafted on to academic medicine. The changing face of academic clinical medicine places significant stresses on the traditional tenure system and raises the question of whether it is fundamentally irrelevant. Tenure has been defended as the protector of free inquiry, insurer of academic excellence, contributor to research and its benefits to society, and a means of selecting the best and brightest for our faculties. Tenure has been criticized as a contributor to academic sloth, codifier of gender and racial discrimination, and as a system unable to deal with the economic realities of high-priced referral specialties. As academic medical centers increase their efforts at clinical outreach, it is unclear whether traditional promotion and tenure will be of interest to clinicians employed solely to deliver primary care. In this special report, I review the history of tenure, arguments for and against its use for clinicians, and proposals for reform. PMID- 7481979 TI - Two biochemical markers effectively used to separate smokeless tobacco users from smokers and nonusers. AB - Data from asbestos workers were used to devise a cutpoint classifier to identify subjects as Nonuser (non-tobacco user), Smokeless (exclusive smokeless tobacco user), and Smoker (ignited tobacco user). In some clinical trials and smoking cessation programs, Smokeless should be separated from Smoker. One therefore needs a marker for smoke exposure, such as thiocyanate, since nicotine levels, as measured by cotinine, could be similar in both groups. Levels of cotinine (ng/mL) and thiocyanate (mumol/L) levels (mean +/- SD) were, respectively: 320.9 +/- 201.1 and 145.9 +/- 63.7 for the Smoker group; 339.1 +/- 327.5 and 32.0 +/- 16.9 for the Smokeless group; and 0.6 +/- 2.6 and 58.2 +/- 33.2 for the Nonuser group. For Nonuser, Smokeless, and Smoker, respectively, the self-reported status was 45.1%, 10.8%, and 44.1%, which was adjusted to 42.2%, 11.6%, and 46.2%; the classifier yielded sensitivities of 100%, 76.1%, and 92.2%; specificities of 96.1%, 97.6%, and 96.4%; and predictive values of 94.9%, 80.6%, and 95.6%. The classifier successfully identified Nonusers, separated Smokeless from Smoker, and determined the prevalence of false reports in our cohort. PMID- 7481980 TI - Discovery method and stage of breast cancer in two different patient populations. AB - Our purpose was to determine the discovery method and stage of newly discovered breast cancers at two different medical centers. To do this, we conducted a retrospective chart review at our urban and suburban teaching hospitals during 1991 through 1992 and 1992, respectively. Only 29 (26%) of 112 cancers at our urban hospital were discovered mammographically; 38 (38%) of 100 neoplasms were discovered using mammography at our suburban hospital. At both institutions, the mammographically discovered cancers had a lower stage than those detected clinically. Despite proven benefits and the efforts of organizations such as the American College of Radiology and the American Cancer Society, screening mammography remains underused in the patient populations that we studied. PMID- 7481981 TI - Delayed development of mid-ileal conduit stenosis: the importance of life-long urologic follow-up. AB - Mid-ileal conduit stenosis is an unusual yet morbid complication following ileal conduit urinary diversion. We report the cases of four patients who developed mid ileal stenosis at an average interval of 19.5 years after urinary diversion, with the longest interval being 25 years. This series emphasizes the importance of life-long follow-up of patients who have had urinary-intestinal diversion. PMID- 7481982 TI - Evaluation of a program to remove barriers to public health care for women with HIV infection. AB - To facilitate access to care for HIV-infected women, a maternal-child program was started at an HIV outpatient clinic in New Orleans. The program consisted of free child care and transportation, separate waiting and examination rooms, female care providers, merged visits for mother and child, and on-site colposcopy services. This study evaluates the efficacy of the program on improving attendance at the clinic by follow-up of 143 women and 557 men (serving as controls). Multivariate models were adjusted for history of intravenous drug abuse, race, age, CD4 cell count, staging of disease, and time in the clinic. Half of the clients achieved high attendance rates. Women were as likely as men to achieve high attendance rates before the interventions, but more likely to do so at both 6 months and 1 year postintervention (after adjustment for the other variables). These results suggest that responding to special gender-related needs can improve women's attendance for clinic visits. PMID- 7481983 TI - Thrombotic tendencies and correlation with clinical status in patients infected with HIV. AB - Previous publications have described thrombotic events with unclear causes in individuals infected with the human immunodeficiency virus (HIV). We stratified the cases of 52 individuals infected with HIV by degree of immunosuppression and the presence of complicating illnesses. Plasma from these individuals was screened for abnormalities that might predispose to thromboses. We found statistically significant differences between patients with CD4 counts < 200/mm3 and those whose CD4 counts were > 400/mm3 in the following: d-dimers, functional protein C, antigenic protein C, total protein S antigen, free protein S antigen, C4b-binding protein (C4b-BP), and von Willebrand antigen (vWD). Free protein S correlated inversely with C4b-BP; vWD directly with total protein S; and protein C inversely with d-dimers. D-dimers were significantly elevated only in immunosuppressed patients with complicating neoplastic/inflammatory disease. We propose that low-grade disseminated intravascular coagulopathy in severely immunosuppressed individuals with HIV and infectious, inflammatory, or neoplastic complications is responsible for depressed protein C, which, together with elevations in total protein S and vWD (markers of endothelial injury), indicates a thrombotic predisposition. PMID- 7481984 TI - Sequential endoscopic/laparoscopic management of sickle hemoglobinopathy associated cholelithiasis and suspected choledocholithiasis. AB - We reviewed the medical records of 17 patients with sickle hemoglobinopathy associated cholelithiasis who were candidates for laparoscopic cholecystectomy (LC) between 1991 and 1994. Eight patients with suspected choledocholithiasis (CDL) were identified, all of whom had preoperative endoscopic retrograde cholangiopancreatography (ERCP), which revealed CDL in 3 patients (18%), all of whom had endoscopic ductal clearance. Choledocholithiasis was suspected because of hyperbilirubinemia or serum liver enzyme abnormalities. Incremental hyperbilirubinemia exceeding "baseline" values by > 5 mg/dL was the best predictor of CDL. Subsequent LC was successful with discharge within 2 days of LC in all but one patient, who was converted to open cholecystectomy. This small series suggests that in sickle hemoglobinopathy patients with cholelithiasis (1) CDL is relatively common among patients with an increase above baseline hyperbilirubinemia, (2) bile duct dilatation, alkaline phosphatase, and serum aminotransferase levels are poor predictors of CDL, and (3) sequential endoscopic/laparoscopic management of cholelithiasis and suspected CDL is successful. PMID- 7481985 TI - Equal effectiveness of very-low-intensity anticoagulation and standard low intensity anticoagulation: a pilot study. AB - We compared the efficacy of very-low-intensity oral anticoagulation (OA) with that of the recommended standard low-intensity oral anticoagulation, using international normalized ratios (INRs). We enrolled 101 patients into a pilot study--51 patients in the very-low-intensity anticoagulation arm (INR 1.4 to 2.0) and 50 in the standard low-intensity anticoagulation arm (INR 2.0 to 3.0). They were monitored for thrombotic/embolic and hemorrhagic complications for an average follow-up of 1.5 years. Two thrombotic/embolic events occurred in the very-low-intensity group; no thrombotic/embolic events occurred in the standard low-intensity group. No major bleeding occurred in the very-low-intensity group; one major hemorrhagic event occurred in the standard low-intensity group. These findings did not achieve a statistically significant difference in major complications between the two groups. It appears that very-low-intensity OA (INR 1.4 to 2.0) is as effective in preventing thromboses as standard low-intensity OA (INR 2.0 to 3.0). PMID- 7481986 TI - Hepatobiliary cystadenoma. AB - Hepatobiliary cystadenomas are rare, truly benign cystic epithelial tumors with the potential to become cystadenocarcinoma. The initial symptom is vague, nonspecific abdominal pain, frequently in middle-aged women, but patients may be asymptomatic. Abdominal ultrasonography may be diagnostic. Intracystic fluid, obtained by sonographically guided fine-needle aspiration, may contain elevated levels of carcinoembryonic antigen. Complete resection or enucleation is the treatment of choice. We present the cases of two patients with hepatobiliary cystadenoma. PMID- 7481987 TI - Advanced triploid pregnancy and preeclampsia. AB - Triploidy, though unusual in advanced gestations, may be associated with preeclampsia. The increase in paternal genetic material associated with the triploid diandric placenta may support the role of immunologic factors in the development of preeclampsia. PMID- 7481988 TI - Tumoral fibrosis and lack of circumscription in granular cell tumor of the breast. AB - Granular cell tumors are benign neoplasms of putative Schwann cell origin that most commonly occur in the tongue. While granular cell tumors in the breast are uncommon, the occurrence of this tumor at this site warrants special consideration, since the mammographic, gross, and microscopic appearance can mimic carcinoma. We describe the case of a 61-year-old white woman with a granular cell tumor of the left breast. Our case and a review of the international literature show that desmoplasia and lack of circumscription are common features of granular cell tumor of the breast. PMID- 7481989 TI - Pulmonary zygomycosis after allogeneic bone marrow transplantation. AB - The Zygomycetes are uncommon human pathogens. They cause illness in some patients who are immunosuppressed and can present as any of several syndromes, including rhinocerebral, pulmonary, gastrointestinal, or cutaneous disease, or as disseminated infection. Pulmonary zygomycosis can occur in any of several patterns and can mimic more common pneumonic processes. This mimicry, as well as the rarity of the disease, may delay diagnosis. We present a case of pulmonary zygomycosis that occurred in a patient who was a bone marrow transplant recipient. The case illustrates several of the common features of the disease in this patient group. PMID- 7481990 TI - Massive edema of the ovary associated with androgenic manifestations. AB - Massive ovarian edema is a rare tumor-like condition of the ovary characterized by marked enlargement of one or both ovaries due to marked accumulation of edema fluid in the ovarian stroma. This paper reviews the literature on massive ovarian edema and presents a case associated with androgenic manifestations. PMID- 7481991 TI - Management of disseminated intra-abdominal lymphangiomatosis with protein-losing enteropathy and intestinal bleeding. AB - We encountered an unusually severe case of intra-abdominal lymphangiomatosis associated with protein-losing enteropathy and intestinal bleeding. A low-fat diet effectively raised the patient's serum levels of hemoglobin and the total serum protein, perhaps by inducing a reduction in intestinal lymph flow and pressure. PMID- 7481992 TI - Fatal outcome of jugular vein cannulation. AB - We describe the case of a woman who had cannulation of the internal jugular vein for the placement of a hemodialysis catheter. The procedure was unsuccessful and resulted in a retropharyngeal hematoma, which in turn caused severe compression on a carotid artery that was already severely narrowed by atherosclerosis. This resulted in massive cerebral hemispheric infarction and death. During the course of hospitalization, ultrasonography, computed tomography, and angiographic studies were done. Cannulation of neck vessels in patients with bleeding diatheses must be done with caution, especially in the presence of severe stenotic disease of the carotid arteries. PMID- 7481993 TI - Basal cell carcinoma due to radiotherapy for carcinoma of the cervix. AB - We report a case of basal cell carcinoma arising in a previously radiated field after treatment of cervical cancer. Our search of the literature yielded no other case of basal cell carcinoma due to irradiation for cervical carcinoma. PMID- 7481994 TI - Seabather's eruption. AB - Seabather's eruption is an unusual rash that develops in individuals who have been swimming in the ocean. We report the case of a 25-year-old woman who had the rash in a typical bathing suit distribution. Several species of cnidarian larvae have been implicated in causing the disease. Symptomatic treatment is the mainstay of therapy for this self-limited rash. Preventive measures allow patients to avoid the disease altogether. PMID- 7481995 TI - Leaking gallbladder remnant with cholelithiasis complicating laparoscopic cholecystectomy. AB - Since its introduction in 1989, laparoscopic cholecystectomy has rapidly become the preferred alternative to open cholecystectomy for symptomatic cholelithiasis. Although possible complications of laparoscopic cholecystectomy are essentially the same as for open cholecystectomy, we report a case of an inadvertent subtotal cholecystectomy, a complication that we believe has not been previously reported. PMID- 7481996 TI - Calcifying amyloidoma of the breast. AB - In this case, a distinctive amyloid tumor of the breast clinically simulated carcinoma, although the patient related onset to trauma. Hepatosplenomegaly, elevated globulins, and anemia led to identification of large amounts of monoclonal IgG-kappa production. The patient died of renal failure within several months despite chemotherapy. The matrix of the breast tumor was tinctorially characteristic of amyloid light chain (AL) protein. The mass contained islands of plasma cells that morphologically suggested local production of amyloid matrix. Moreover, plasma cell and matrix immunohistochemically displayed reactivity of IgG-kappa protein, indicating a clonal plasma cell infiltrate. Pseudo-acinar arrangement of plasma cells may be misinterpreted as epithelial cells in needle biopsy specimens. The notion that some amyloidomas may represent in situ production of protein by clonal immunocytes ("secretory immunocytomas") should be further studied. PMID- 7481997 TI - Unilateral polycystic (dysgenetic) disease of the parotid gland. AB - We report a case of unilateral polycystic disease of the parotid gland. Only eight cases of this disease have previously been published in the English language literature, and seven of them were bilateral. Thus, we are reporting the second case of unilateral involvement. The disease is apparently limited to the parotid gland and to women. Clinically, a fluctuating, long-standing, nontender parotid gland swelling is usually noticed in adulthood. Histologically, there are numerous epithelial-lined cysts, which appear to be derived from intercalated ducts. This disease probably represents a developmental condition. PMID- 7481998 TI - Systemic lupus erythematosus in a child with sickle cell disease. AB - Although sickle cell disease (SCD) and systemic lupus erythematosus (SLE) are two distinct chronic diseases, many clinical features are common to both conditions. We describe a young patient who had a mild clinical course of SCD until SLE developed when he was 15 years old. His initial manifestations of SLE including fever, chest pain, and lung infiltration with pleural effusion were thought to be complications of SCD. However, a deteriorating clinical course, presence of facial and truncal rash, and persistent pleural effusion led to the diagnosis of SLE. We compare our case and the 10 previously reported cases and discuss the possible association of complement defects and the pathogenesis of SLE in patients with SCD. Our report illustrates the importance of considering other disease processes when clinical features are atypical of SCD. PMID- 7481999 TI - Treatment of a pleural effusion due to Charcot's arthropathy. AB - Cervical spinal cord injury compromises the ventilatory muscles and is associated with respiratory complications. Patients who are quadriplegics are also at increased risk of Charcot's arthropathy of the spine. The development of a large pleural effusion in quadriplegia further increases respiratory compromise. I present a case of Charcot's arthropathy of the spine causing a large pleural effusion that presented as respiratory distress in a quadriplegic. Pleurodesis was successful in preventing reaccumulation of the pleural effusion with a reduction in dyspnea. PMID- 7482000 TI - Imported fire ants--from medical nuisance to medical menace: new concerns for medical facilities in endemic areas. PMID- 7482001 TI - Biliary sludging in trauma patients. PMID- 7482002 TI - D-dimer test for HELLP syndrome. PMID- 7482003 TI - Hereditary multiple glomus tumors (glomangiomas) PMID- 7482004 TI - An economic evaluation of schizophrenia--1991. AB - In 1991, the costs for schizophrenia, which has a lifetime prevalence of 1.5% among adult Americans, totaled $65 billion. Costs were broken down into their direct and indirect components. Direct costs, which totaled $19 billion dollars, consisted of treatment-related expenditures such as those for inpatients and outpatients, as well as nontreatment-related expenditures such as those for the criminal justice system used by individuals with schizophrenia. The direct costs were fairly similar to those of other recent estimates of the cost of schizophrenia. Indirect costs, which were $46 billion dollars, included the lost productivity of both wage earners ($24 billion) and homemakers ($4.5 billion), individuals who were in institutions ($4.5 billion) or who had committed suicide ($7 billion), and caregivers who took care of schizophrenic family members ($7 billion). Our method for calculating the indirect costs was slightly different than methods used in prior studies, which may account for our estimates being higher. The method for determining each expenditure is provided, and the implications of these staggering costs are discussed. PMID- 7482006 TI - An economic evaluation of manic-depressive illness--1991. AB - In 1991, the costs for manic-depressive illness, which has a lifetime prevalence of 1.3% among adult Americans, totaled $45 billion. Costs were broken down into their direct and indirect components. Direct costs totaling $7 billion consist of expenditures for inpatient and outpatient care, which are treatment related, as well as nontreatment-related expenditures such as those for the criminal justice system used by individuals with manic-depressive illness. Indirect costs, which were $38 billion, include the lost productivity of both wage-earners ($17 billion) and homemakers ($3 billion), individuals who are in institutions ($3 billion) or who have committed suicide ($8 billion), and caregivers who take care of manic-depressive family members ($6 billion). The method for determining each expenditure is provided, and the implications of these staggering costs are discussed. These calculations rely heavily on methods and data bases that were developed for the accompanying paper on the costs of schizophrenia. PMID- 7482005 TI - Schizophrenia: shifting the balance of care. AB - Schizophrenia poses a major challenge to policy makers in health and social care in England. As deinstitutionalisation has progressed, public concern about people with the condition has increased owing to a small number of public incidents. This paper describes the overall balance of care or support for people with schizophrenia, particularly between different settings and the services received. Using these data we are able to provide a basic estimate of the current resource implications of schizophrenia care in England with respect to different public sector agencies, the voluntary sector and service users and their families. The targeting of public sector resources between different care settings is also considered. The discussion suggests that resources are still targeted at existing care facilities rather than on individuals, and examines the roles of individual care agencies and the relationships between them. Four broad resource guidelines are suggested that could improve the effective use of resources for schizophrenia care. PMID- 7482007 TI - Managed mental health care and patterns of inpatient utilization for treatment of affective disorders. AB - In this analysis we made use of a large data base of individuals insured by large American corporations to estimate the impact of managed care provision on hospital care for depression. Data on 6,348 individuals hospitalized for depression were examined to assess the effect of managed care techniques on the cost per episode and the likelihood of rehospitalization. Preadmission certification programs were found to lead to significant long- and short-run savings for payers. PMID- 7482008 TI - Depressive disorders: treatment patterns and costs of treatment in the private sector of the United States. AB - The purpose of this paper is to examine the treatment patterns and costs of treatment for depressive disorders in the private sector of the United States. Based on the 1987-1989 calendar year MEDSTAT claim data, 40,898 patients were identified with a principal diagnosis of depressive disorder. Among a list of CPT 4 code procedures, individual psychotherapy had the highest frequency of usage followed by individual visits. Compared to individual psychotherapy, group/family psychotherapy had a much lower frequency of usage. Very few diagnostic episodes had laboratory work. In inpatient settings, costs of physician procedures and laboratory services were 2 times greater for patients with major depression or bipolar disorder than for patients with depression not otherwise specified (NOS) or dysthymic disorder. As expected, costs varied widely per episode. As the severity of illness increased, the cost variation became wider. PMID- 7482009 TI - Crisis intervention and affective disorders: a comparative cost-effectiveness study. AB - In recent years, psychiatric institutions have been increasingly urged to justify their clinical policies in order to ensure both effective treatment and efficient management. Assessment instruments for effectiveness and costs are essential to respond to these needs. The aim of this study was to determine the cost effectiveness of treatments for major depressive disorders. We conducted a comparative pilot investigation of treatment costs in patients with a major depressive episode assigned to specialised out-patient crisis intervention, to specialised in-patient treatment and to standard mental hospital care. The study included 122 subjects. The inclusion criteron was a diagnosis of DSM-III-R major depressive episode. Costs were assessed by determining the average cost for each treatment and the modalities of payment systems. Treatment duration and costs were high, but specialised crisis intervention may considerably reduce the duration of hospitalisation and its associated costs. The average costs of treating major depression were about 4 times greater in the specialised hospital unit than in the standard hospital unit and the crisis intervention centre. The burden of payment was comparatively higher for the state and reduced for insurance companies when the treatment of major depressive disorders involved less in-patient care. PMID- 7482010 TI - The role of private insurance in financing treatment for depression. AB - The health care system in the United States is expensive but fails large groups of its citizens, particularly those with serious mental illness. This paper examines the resource implications of adequate treatment for depression. An analysis of the various tasks necessary for a proper treatment program is used to indicate places where the current mechanisms of provision fall short. Appropriate innovations in principles of funding are suggested. PMID- 7482011 TI - Effectiveness, patient matching, and insurance coverage for depression. AB - The effectiveness of treatment for depression is not in doubt, and common humanity would indicate that it should be provided to those that need it. However, in this paper I argue that the existence of effective treatment should not be regarded as a sufficient condition for the extension of insurance cover. This is because insurance coverage cannot guarantee that treatment is delivered effectively, nor that it is given to the right people; neither does it lend itself to the maximization of cost-effectiveness. Payment systems must take account of these considerations. PMID- 7482012 TI - Isometric and dynamic performances of the trunk and associated factors. AB - STUDY DESIGN: Static and dynamic performances of the trunk were measured on an isoinertial device and data were collected regarding the working conditions and the anthropometric and psychosocial characteristics of the workers. OBJECTIVES: To describe the difference in static and dynamic performances of the trunk between workers with and without low back pain and to investigate the anthropometric, personal, and occupational factors associated with these performances. SUMMARY OF BACKGROUND DATA: The isometric torque and the parameters of displacement, velocity, and torque during dynamic tests were chosen to describe the isoinertial performances of the trunk. Dynamic characteristics in particular were investigated because it has been suggested in the literature that these discriminate best between workers with and without low back pain. METHODS: The protocol included a 52-item questionnaire, a clinical examination with a series of anthropometric measurements, and the isometric and dynamic tests (against 25% and 50% of the isometric values) on the isostation. RESULTS: The variables significantly associated with the trunk performances were weight, height, muscularity, sports, and leisure activities as positive factors, and linearity, health problems, and vehicle driving as negative factors. When these covariates were taken into consideration, the significant differences between subjects with and without low back pain were related only to the velocities during dynamic tests, which were reduced by 5.9%. CONCLUSIONS: The performances of the trunk are associated mainly with the anthropometric characteristics of the subjects but also with factors such as lumbar status, health, working conditions, and leisure activities. PMID- 7482013 TI - Spinal cord expansion after decompression in cervical myelopathy. Investigation by computed tomography myelography and ultrasonography. AB - STUDY DESIGN: This study analyzed anatomic characteristics of 44 patients with cervical myelopathy. OBJECTIVES: This study was conducted to clarify the patterns of spinal cord expansion after decompression, their relationship to clinical recovery, and the capacity of the compressed spinal cord for restoration. SUMMARY OF BACKGROUND DATA: No previous studies have clearly assessed the relationship between morphological changes of the spinal cord and clinical improvement after posterior decompression in patients with cervical myelopathy using computerized tomography myelography and intraoperative spinal sonography. METHODS: Forty-four patients with cervical myelopathy underwent posterior decompression. The diagnosis was cervical spondylosis in 26 patients and ossification of the posterior longitudinal ligament in 18 patients. The cross-sectional area, sagittal diameter, and frontal diameter of the spinal cord at the level of maximum compression were determined by computerized tomography myelography (performed before and 1 month after decompression) and by intraoperative spinal sonography. RESULTS: Expansion of the spinal cord was defined as immediate or gradual. Of the 44 patients, 20 demonstrated gradual expansion, and their clinical recovery rate was 68.4%. In contrast, the clinical recovery rate of patients without gradual expansion was only 32.6%. Good clinical recovery was significantly related to gradual expansion. The patients were divided into four groups (A-D) based on the clinical recovery rate and compression ratio. Group A (severe compression and poor clinical recovery) showed a significantly smaller cross-sectional area of the spinal cord than Group D (severe compression, but good clinical recovery). CONCLUSIONS: Preservation of the cross-sectional area of the spinal cord, even when it is severely compressed and deformed, is an important factor in a good surgical outcome. PMID- 7482014 TI - Decreased width of the spinal canal in patients with chronic symptoms after whiplash injury. AB - STUDY DESIGN: The sagittal diameter of the cervical spinal canal was measured on standard lateral radiographs of 48 consecutive patients with acute whiplash injury. A follow-up was done 12 months after injury. OBJECTIVES: The purpose of this study was to evaluate the relationship between spinal canal width and chronic symptoms after whiplash injury. SUMMARY OF BACKGROUND DATA: A narrow sagittal diameter of the cervical spinal canal has been correlated with neurologic deficits in degenerative conditions and after fractures. The authors are not aware of previous studies on whiplash patients related to this area. METHODS: Forty-eight consecutive patients were evaluated for whiplash trauma after car accidents. Twenty-four patients (12 men and 12 women; mean age, 37 years) had persistent symptoms after 12 months, while 24 patients (12 men and 12 women; mean age, 34 years) were asymptomatic. All measurements were done from standard lateral radiographs with a graphic digitizer connected to a microcomputer. Three anteroposterior and one oblique measurement of the canal were reconstructed for each vertebrae from C2-C6. Multivariate analysis of variance was used to evaluate any association between gender, remaining symptoms at follow-up, and the recorded values of the spinal canal width. RESULTS: The spinal canal was significantly smaller in the patients with persistent symptoms than in the asymptomatic group. A significant difference also was found between men and women. CONCLUSION: Narrow diameter of the cervical spinal canal is unfavorable in patients with whiplash. PMID- 7482015 TI - An epidemiologic study of episodes of back pain care. AB - STUDY DESIGN: This was a prospective community-based, observational design. OBJECTIVE: To describe the epidemiology and risk/prognostic factors for back pain episodes of care in a population representing the nonelderly in the United States. SUMMARY OF BACKGROUND DATA: Previous United States studies of the epidemiology of back pain care have used defined industrial populations or have relied on the patient's recall of symptoms and care. METHODS: Claims forms from the RAND Health Insurance Experiment, a randomized controlled trial of the use of health services, were analyzed. Claims forms were selected if one of the patient designated reasons for the visit was back pain. Visits were grouped into episodes of care. Descriptive statistics were calculated for episodes. Multivariate logistic regression was used to calculate adjusted odds ratios for independent explanatory sociodemographic and health status variables associated with back pain episodes of care. RESULTS: The 3105 adults in the Health Insurance Experiment had a combined 11,171 person-years of exposure. Six-hundred-eighty-six persons (22%) had a total 1020 episodes of back pain care, representing 8825 visits. Seventy-one percent of persons had a single episode during the Health Insurance Experiment, and 40% of these episodes consisted of a single visit. There were 9.1 episodes per 100 person-years. Insurance status, geographic site, white race, lesser education, poorer physical functioning, and greater pain at base-line all were independently associated with having a back pain episode of care. CONCLUSIONS: Back pain episodes of care occur commonly in the adult U.S. population, but usually are brief and recur infrequently. PMID- 7482016 TI - Early changes in bone mineral density above a combined anteroposterior L4-S1 lumbar spinal fusion. A clinical investigation. AB - STUDY DESIGN: Fifteen patients meeting rigid study entrance criteria for undergoing a combined anteroposterior L4-S1 spinal fusion as a first time operation were included in the study. Bone mineral density changes at adjacent segments (L3 and L2) were assessed by scanning patients in the anteroposterior and lateral projections before surgery and 3 and 6 months after surgery. OBJECTIVES: To prospectively evaluate changes in bone mineral density at adjacent segments after lumbar spinal fusion and postoperative bracing. SUMMARY OF BACKGROUND DATA: The majority of studies investigating changes in bone mineral density after spinal fusion have been performed in canines. Although such work has provided valuable insight, its application to humans is questionable because the loading mechanics are so different. METHODS: Bone mineral density was assessed noninvasively by dual energy x-ray absorptiometry in the lateral and anteroposterior projections. Data were analyzed with special edge detection software. RESULTS: In the lateral projection at L3, one level above the fusion, the preoperative bone mineral density was 0.91 +/- 0.24 (mean +/- standard deviation in g/cm2). This decreased significantly at 3 months to 0.82 +/- 0.24 and remained reduced at 6 months (0.80 +/- 0.21). This reflected a loss of primarily trabecular bone. In the anteroposterior projection at L2, there was a significant loss at 3- and 6-month follow-up (1.24 +/- 0.15 to 1.19 +/- 0.15 and 1.19 +/- 0.14). This may reflect changes at either or both of the anterior and posterior columns. CONCLUSION: Significant decreases in bone mineral density were noted above fused lumbar segments. This may have been related to immobilization or altered mechanics associated with arthrodesis. Trabecular bone of the axial skeleton is most responsive to change, compared with cortical bone, because of the higher metabolic activity. This may explain the reduction of bone mineral density at L3. Changes at L2 may have been related to altered loads at the anterior or posterior columns and should be further investigated. PMID- 7482017 TI - Morphometric study of the lumbar spinal canal in the Korean population. AB - STUDY DESIGN: The anatomic dimensions of the vertebral body and spinal canal of the lumbar spine were analyzed in Koreans. OBJECTIVES: To determine the normal dimension of the lumbar spinal canal in Koreans, to determine whether there are any racial differences in the morphometry of the lumbar spinal canal, and to provide criteria for diagnosing spinal stenosis in the Far Eastern Asian. SUMMARY OF BACKGROUND DATA: Some radiologic and anatomic studies have been conducted regarding the size of the lumbar spinal canal of whites and blacks in western and African countries. METHODS: One-thousand-eight-hundred measurements were performed on the transverse and sagittal diameters of vertebral bodies and spinal canals using complete sets of 90 lumbar vertebrae. RESULTS: The mean mid-sagittal diameter of the lumbar spinal canal in the Korean population was less than that measured in white and African populations, but there was no significant differences between the Korean, white, and African populations regarding the transverse diameter of the lumbar spinal canal. CONCLUSION: The mid-sagittal diameter of the lumbar spinal canal is narrowest in the Far Eastern Asian population; the radiologic criteria of spinal stenosis should be reconsidered for these people. PMID- 7482019 TI - Postnatal development of the thoracic spine. AB - STUDY DESIGN: Forty-two thoracic spines ranging from neonate to 15 years of age were studied. OBJECTIVES: Postnatal development of the thoracic vertebrae and their component elements were cataloged. SUMMARY OF BACKGROUND DATA: This study was part of an ongoing series concerning the progressive morphologic development of the human postnatal skeleton. METHODS: The spines were studied morphologically and radiologically. RESULTS: Fusion of the thoracic spine neurocentral synchondroses occurs in a cranial to caudal direction in humans by 6 to 7 years of age. The transverse processes exhibit a progressive posterior and inferior angulation with age and moving down the thoracic spine. The facet joints angulate accordingly. The spinous processes elongate and increasingly angulate with age, especially in the mid-thoracic region. The relative contribution of the posterior elements, compared with the centrum, to the growth of the vertebral body progressively decreases moving down thoracic spine. CONCLUSION: This study demonstrated areas of anatomic change that undoubtedly play a role in normal and abnormal spine development. PMID- 7482018 TI - Adolescent idiopathic scoliosis and cerebral asymmetry. An examination of a nonspinal perceptual system. AB - STUDY DESIGN: This study analyzed the asymmetry of a nonspinal sensory system, comparing healthy children and those with adolescent idiopathic scoliosis. OBJECTIVES: To test the hypothesis that differences in neurologic functioning that have been observed in idiopathic scoliosis are confined to motor organization and that the etiology thus is a defect solely of the motor cortex. SUMMARY OF BACKGROUND DATA: Recent reports associating scoliosis convexity with equilibrium control, central processing, handedness, and motor lateralization have suggested that idiopathic scoliosis is connected causally with the motor cortex. METHODS: Dichotic listening tests were performed on 20 healthy children and 31 children with adolescent idiopathic scoliosis. This tests perceptual asymmetry, and thus the organization of cognitive processing in the brain, a higher function that is not associated with posture or motor function. RESULTS: Subjects with scoliosis were significantly more lateralized for linguistic processing than the control group, indicating they had a greater degree of left right asymmetry throughout their cortical organization. CONCLUSION: In adolescent idiopathic scoliosis, the organization of the entire brain was more strongly lateralized than in subjects without scoliosis. It is unlikely that this caused or was caused by the spinal curvature. Subjects with adolescent idiopathic scoliosis showed generalized asymmetry of many functions and structures. An examination at the level of morphology and development is proposed. PMID- 7482020 TI - Reduction of bone retropulsed into the spinal canal in thoracolumbar vertebral body compression burst fractures. A prospective randomized comparative study between Harrington rods and two transpedicular devices. AB - STUDY DESIGN: This was a prospective, randomized study. OBJECTIVE: To compare the ability of three methods of internal fixation (Harrington rods, AO internal fixator, posterior segmental fixator) to obtain reduction of intracanal fragments in thoracolumbar vertebral compression burst fractures. SUMMARY OF BACKGROUND DATA: Sixty-seven acute thoracolumbar compression burst fractures of T12 or L1 were randomized into three groups that were treated using one of the three methods. Reduction was accomplished indirectly by distraction applied using the fixation device. METHODS: The spinal canal encroachment was calculated as a percentage of the estimated pre-injury value from serial transverse computed tomographic scans obtained on admission and immediately after surgery. RESULTS: The median preoperative sagittal encroachment of the spinal canal was 37% (range, 0-90%) of the normal diameter. All three methods of internal fixation produced a spinal canal clearance provided that the patient was operated on within 4 days after trauma. The median postoperative encroachment varied from 13% (range, 0 37%) to 22% (range, 0-37%), the best reduction being attained using Harrington rods and the poorest with the posterior segmental fixator. There was a suggestive statistical significance between these two. CONCLUSION: The differences in postoperative spinal canal encroachment and ability to obtain spinal canal clearance observed between the devices studied were small. There seems to be no reason to base the choice of the operative method in thoracolumbar fractures on any hypothetical differences in reductive power between Harrington rods and the AO internal fixator. PMID- 7482021 TI - Long-term follow-up results of thoracolumbar fractures after posterior instrumentation. AB - STUDY DESIGN: This retrospective study examined the hospital records of 60 patients with thoracolumbar fractures treated with posterior fusion and spinal instrumentation. The mean follow-up period was 66 months. OBJECTIVES: The goal of this study was to evaluate and analyze the long-term outcome and socioeconomic conditions of patients who had sustained a thoracolumbar fracture. SUMMARY OF BACKGROUND DATA: In four patients, additional spinal injuries were detected. Three of the patients evaluated with magnetic resonance imaging showed cord contusion and edema. In six patients, ruptured disc fragments were detected by preoperative magnetic resonance imaging or during surgery. METHODS: Long-term follow-up results in 60 patients with unstable thoracolumbar fractures treated with posterior fusion and spinal instrumentation were analyzed. Neurologic outcomes and independence in function and daily living activities were reviewed. Age, sex, mechanism of injury, associated injuries to the spinal cord, and associated injuries to the spinal cord and other systems were analyzed. Fractures were classified according to the system of Ferguson and Allen. RESULTS: The patients with incomplete spinal cord injury showed significant functional improvement. During the follow-up period, 28 patients showed neurologic improvement. Postoperative complications occurred in 11 patients. Five patients required late rod removal because of rod dislocation. CONCLUSION: Regardless of neurologic recovery, most patients reported some disability, usually caused by pain. Inability to return to alternative jobs resulted from insufficient educational background rather than neurologic dysfunction. Advanced academic achievement was the single most important predictive factor of ability to return to work. PMID- 7482022 TI - Latent solitary tuberculous psoas abscess 52 years after healed thoracolumbar tuberculous spondylitis. AB - STUDY DESIGN: This study reports on an extremely rare case of tuberculous psoas abscess and describes the mode of diagnosis and treatment. OBJECTIVE: This patient is presented to emphasize that cases of solitary psoas abscess resulting from tuberculosis exist today. SUMMARY OF BACKGROUND DATA: No recent cases of latent solitary tuberculous psoas abscess have been reported, to the authors' knowledge. METHODS: A tuberculous psoas abscess associated with fistula to the greater trochanter may remain dormant for years after tuberculous spondylitis has healed, as a distinct entity without concomitant active bone infection. In our 58 year-old female patient, the diagnosis of a psoas abscess was greatly aided by the "three pass" technetium bone scan and computed tomography, but the anamnesis also was important. RESULTS: Anti-tuberculosis medication (streptomycin, aminosalicylic, and isoniazid) combined with open drainage, curettage of the psoas, and simultaneous revision of the fistula eradicated the disease, making the course of the disease uneventful until the 5-year follow-up evaluation. CONCLUSIONS: Orthopedic surgeons must be aware of the rare, delayed appearance of a solitary psoas abscess after tuberculous spondylitis. PMID- 7482023 TI - Percutaneous intradiscal radio-frequency thermocoagulation. A cadaveric study. AB - STUDY DESIGN: This study examined the effect of radio-frequency thermocoagulation on cadaveric disc material. Radio-frequency lesions are induced to decompress the nerve root by coagulating the nucleus pulposus. OBJECTIVES: The purpose of this study was to assess the potential of the radio-frequency lesion technique for treating lumbar disc herniation percutaneously. The effect of radio-frequency thermocoagulation on cadaveric disc material was studied. SUMMARY OF BACKGROUND DATA: An intradiscal bipolar electrode is used for the thermocoagulation of the nucleus pulposus. METHODS: Lumbar discs of 27 adult cadavers were used for the experiments within 72 hours of donors' deaths. The physical and thermal parameters of intradiscal radio-frequency thermocoagulation were determined experimentally. Acute macroscopic and histologic changes before and after thermocoagulation were investigated. The temperature distribution of the tissue surrounding the disc was measured during the operation using thermocouples to evaluate potential heat damage. Also, computed tomography and magnetic resonance imaging of the disc were performed before and after thermocoagulation. RESULTS: Macroscopic observation showed that the lesion induced by radio-frequency thermocoagulation is homogeneous and intense without necrosis, and is limited to the nucleus pulposus. Histologically, the endplates and vertebral bodies were unaffected and an architectural disorganization of the stroma of the nucleus pulposus was seen. The temperature increase of the tissue surrounding the discs did not exceed 3-4 C. CONCLUSION: A bipolar electrode and a radio-frequency alternating current permits percutaneous intradiscal thermocoagulation for destruction of a portion of disc material in a cadaveric spine. PMID- 7482024 TI - Applications of the finite element method to thoracolumbar spinal research--past, present, and future. AB - The finite element method has been used in spine biomechanics research for nearly a quarter of a century. Recent developments have made it possible to simulate a variety of clinical situations in an increasingly realistic manner, and have elevated the finite element method into a fully complementary partnership with experimental approaches for investigating clinical problems of the spine. The impact of several of these new developments on present and future spine biomechanics research is addressed in this update. PMID- 7482025 TI - Vittorio Putti. PMID- 7482026 TI - Multilevel cervical spondylosis. Laminoplasty versus anterior decompression. AB - Poor overall outcome and a high incidence of postoperative kyphosis and progressive myelopathy have driven surgeons away from decompressive laminectomy as a treatment for multilevel cervical spondylosis. Dr. Henry Bohlman advocates anterior decompression and fusion as the best approach to the pathophysiology of this disorder, while Dr. Kiyoshi Hirabayashi believes that laminoplasty represents an excellent strategy for patients with degenerative disease, as well as those with ossification of the posterior longitudinal ligament. PMID- 7482027 TI - The virtual hospital. Providing multimedia decision support tools via the Internet. AB - Physicians in the clinical setting remain isolated from important sources of medical information. The authors have created a multimedia database known as The Virtual Hospital that improves access to current medical data, which is used to improve patient care decisions. The Virtual Hospital is a digital health sciences library stored on a server (computer) at The University of Iowa and delivered via the Internet to inexpensive personal computers in the workplace. The emerging standard of the World Wide Web is used to provide cross-platform distribution. PMID- 7482028 TI - Flexion-relaxation phenomenon (transfer of loads between lumbar tissues during the flexion-relaxation phenomenon) PMID- 7482029 TI - Thoracic discography: a safe and reliable technique. PMID- 7482030 TI - Deletion mapping of highly conserved transcribed sequence downstream from APRT locus. AB - To investigate the nature of DNA sequence rearrangements occurring in a highly malignant human colorectal carcinoma cell line (SW620) exhibiting a high level of chromosome instability, we characterized the molecular basis of deletions eliminating APRT. Deletions in SW620 resembled those in a variety of cell lines. They were joined at regions of little similarity through mono-, di-, or trinucleotide repeats. Breakpoint regions were rich in di- and trinucleotide repeats that might constitute pause sites for the replication complex. Deletions ranged in size from 1.8 to approximately 70 kb and were "directional" in that they eliminated sequences upstream of APRT but not downstream. Analysis of downstream sequences suggested that this pattern of deletion was due to the presence of another gene. Transcripts from these two genes converged but did not overlap. Given that this gene was not deleted in any hamster or human mutants, it appears essential for cell viability. This organization has important consequences for the pattern of mutation and repair of this region. PMID- 7482031 TI - UMP synthase activity expressed in deficient hamster cells by separate transferase and decarboxylase proteins or by linker-deleted bifunctional protein. AB - Segments of the human UMP synthase cDNA coding for the orotate phosphoribosyl transferase (OPRT) and orotidylate decarboxylase (ODC) domains of the bifunctional protein UMP synthase were produced by polymerase chain reaction techniques and cloned into a eukaryotic expression vector. The separate OPRT and ODC vectors, along with a selectable marker, were cotransfected into UMP synthase deficient hamster cells (Urd-C) that require exogeneous uridine for growth. Transfected Urd-C cells surviving selection in media without added uridine were isolated and designated transferase decarboxylase Urd-C (TDU). All of the selected colonies contained DNA corresponding to the OPRT and ODC expression vectors. Two cell lines (TDU3 and TDU5) integrated many more copies of the OPRT and ODC vectors into their genomes compared to the other TDU lines. A 28.6-kDa ODC protein band and a 24.4-kDa OPRT band were detected on western blots with UMP synthase-specific polyclonal antiserum. The OPRT activity of the TDU lines was up to 8.7 times the OPRT activity of control CHL cells, and the ODC activity was up to 12.5 times control levels. Both OPRT and ODC activities in the monofunctional proteins were less heat stable than in the bifunctional UMP synthase protein. The monofunctional OPRT protein was less stable than the ODC protein at 45 degrees C. Growth of transfected cells in 6-azauridine resulted in striking increases in activity and temperature stability for the monofunctional ODC protein. A UMP synthase bifunctional protein was constructed with a deletion of the suspected linker region joining the two catalytic domains. The linker-deleted UMP synthase showed no significant change in either OPRT or ODC activity or temperature stability. The increased stability of the bifunctional protein may be a factor in its evolutionary selection in mammalian cells. PMID- 7482032 TI - Severe phenotype in mice with termination mutation in exon 2 of cystic fibrosis gene. AB - Mice with a termination codon mutation in exon 2 of the cystic fibrosis (CF) gene were generated using homologous recombination in embryonic stem cells. Animals homozygous for the mutant allele display a severe intestinal phenotype similar to that previously reported for CF mutant mice. The null nature of this allele was demonstrated by the absence of detectable wild-type mRNA, by the absence of detectable CFTR in the serous gland collecting ducts of salivary tissues, and by the lack of cAMP-mediated short-circuit current responses in colonic epithelium of mutant animals. PMID- 7482033 TI - Mevalonate regulates polysome distribution and blocks translation-dependent suppression of 3-hydroxy-3-methylglutaryl coenzyme A reductase mRNA: relationship to translational control. AB - We reported previously that 3-hydroxy-3-methylglutaryl coenzyme A reductase synthesis is regulated at the translational level by mevalonate. To determine at what stage mevalonate affects reductase synthesis, we examined the distribution of reductase mRNA in polysomes from cells treated with lovastatin alone; lovastatin and 25-hydroxycholesterol; or lovastatin, 25-hydroxycholesterol, and mevalonate. In lovastatin-treated cells, reductase mRNA was primarily associated with heavy polysome fractions. When 25-hydroxycholesterol was added to lovastatin treated cells, reductase mRNA levels were reduced approximately fourfold in all polysome fractions, with no accompanying redistribution of reductase mRNA into lighter polysome fractions. However, addition of both 25-hydroxycholesterol and mevalonate to lovastatin-treated cells shifted reductase mRNA from heavier to lighter polysome fractions. No change in the distribution of control beta-actin or ribosomal protein S17 mRNA occurred with any of the treatments. These results suggest that mevalonate suppresses reductase synthesis at the level of initiation. When the translation inhibitor cycloheximide was added to all three regimens, reductase mRNA shifted into heavy polysome fractions. Treatment with either lovastatin alone or lovastatin plus 25-hydroxycholesterol resulted in a 50% greater loss of reductase mRNA from the heavy polysome fractions compared to the same fractions from noncycloheximide-treated cells. No loss of reductase mRNA occurred when cycloheximide was added to cells treated with both 25 hydroxycholesterol and mevalonate. beta-Actin mRNA levels and polysome distribution were not significantly changed by cycloheximide under any of these conditions. Translationally mediated suppression of reductase mRNA did not occur when protein synthesis was inhibited with puromycin. Our results indicate that regulation of reductase mRNA levels is translation-dependent and is linked to the rate of elongation. PMID- 7482034 TI - Chimeric envelope glycoproteins constructed between amphotropic and xenotropic murine leukemia retroviruses. AB - A set of chimeric envelope proteins between amphotropic and xenotropic murine leukemia retroviruses (MuLV), two closely related members in the MuLV family, were constructed. The purpose was to examine the regions that could be successfully exchanged between these two similar viral envelope proteins. The data indicate that fully active chimeras can be built when the junction is either at the EcoRI site (amino acid 169) 42 amino acids N-terminal to the polyproline hinge of gp70 (named CH4) or at the ScaI site (aa 593) in the membrane spanning portion of p15E (CH1). However, a chimera at the AflII site (aa 125, CH5) and two in the C-terminal end of gp70 (aa 418, CH2; aa 326, CH3) were inactive. These results, taken together with other data from our laboratory and others, suggest that the entire gp70/p15E structure is sensitive to alterations and that even envelope proteins that are very similar have only a limited ability to exchange sequences. PMID- 7482037 TI - [DSA of the pelvic-leg blood vessels with Imeron 400 and Optiray 350. Does a higher iodine delivery rate provide better vascular imaging?]. PMID- 7482036 TI - [Musculoskeletal tumors: epidemiology, clinical aspects and diagnostic requirements]. PMID- 7482038 TI - [The postmyelography syndrome--very rare in use of an "atraumatic needle". A controlled double-blind study]. PMID- 7482035 TI - Evidence for gene silencing by DNA methylation in normal human diploid fibroblasts. AB - Human diploid fibroblasts, strain MRC-5, were permeabilized by electroporation and treated with 5-methyl deoxycytidine triphosphate (5-methyl dCTP) in the S phase of the cell cycle. The frequency of TGR HPRT- cells was increased up to 20 fold in comparison to control untreated cultures. Representative TGR clones were unable to grow in HAT, and these were treated with 5-azacytidine (5-aza-CR). In many cases subsequent growth in HAT medium was observed, but in others it is likely that the cells had run out of growth potential. The results provide the first evidence of the silencing and reactivation of a gene in normal diploid mammalian cells. PMID- 7482039 TI - [Radiotherapy of advanced esophageal carcinoma--results from the Munster clinic]. PMID- 7482040 TI - [Postoperative, conventional fractionated radiotherapy for prevention of heterotopic ossification after total endoprosthesis of the hip joint]. AB - Radiotherapy is the only local means of treatment in the prevention of heterotopic bone formation. It is possible to reduce the incidence of HO following total hip arthroplasty from an average of 50% without further treatment to an average of 8% using different doses and fractionation schemes. In this retrospective clinical study 22 hips of 20 patients were treated with a conventional fractionated radiotherapy. Total doses of 12.0-20.0 Gy in daily fractionation of 200 cGy single doses were applied 18 hips were postoperatively irradiated as secondary prophylaxis against recurrence of HO. 4 hips were high risk cases with contralateral HO after previous hip surgery. After 6-30 months observation periods the roentgenogramms were analyzed with a modified Brooker Score. Patients with a score > II were classified as nonresponders. 95.5% of the treated hips showed satisfactory results, only one hip (4.5%) achieved a score IIIA. In this case radiotherapy was interrupted after 3 x 200 cGy. The high therapeutic efficiency of local irradiation in the prevention of heterotopic bone formation following total hip replacement could be proved. No considerable side effects were observed. Further prospective clinical investigations are necessary to optimize the doses and fractionation schemes. PMID- 7482041 TI - [The physician-patient relationship and sophisticated modern technology]. PMID- 7482042 TI - [High resolution CT of the petrous bone for locating a foreign body in the middle ear]. PMID- 7482043 TI - [Computerized tomography signs of shoulder instability]. PMID- 7482044 TI - [Lipoma of the stomach--diagnosis and therapy]. PMID- 7482045 TI - [Pulmonary complications in HIV-positive patients with minimal immune deficiency]. PMID- 7482046 TI - [Computerized tomography measurement of lung ventilation by inhalation of Isovist 300]. PMID- 7482047 TI - [Usefulness and risk of roentgen diagnosis. A contribution to quality assurance]. PMID- 7482048 TI - [Charles Thurstan Holland (1863-1941)]. PMID- 7482049 TI - [Surgery of infectious complications in prostheses in the abdominal aorta]. AB - The authors describe the successful solution of a serious and feared complication, i.e. infection of a vascular prosthesis. A hitherto not used procedure was adopted-concurrent removal of the infected prosthesis and restoration of the blood flow into both extremities by an extra-anatomical axillobiiliac bypass. The authors draw attention to the advantages of this method in context with possible future solutions; and with regard to the hitherto not complicated postoperative course they assume that this type of treatment could be final. At the same time they draw attention to the complexity of the problem from the diagnostic and therapeutic aspect. PMID- 7482050 TI - [Tracheal replacement with a swine prosthesis]. AB - In 12 experimental operations the authors tested the biocompatibility of a xenologous trachea from a pig which was used to replace a 5 cm defect of the trachea in a dog. After special preparation eliminating the antigenicity of the pig trachea the thus prepared graft was sutured into the thoracic portion of the dog trachea. The experiment could not be evaluated in two instances because the dog died on the operating table. In the remaining ten cases it was found that all anastomoses healed perfectly. The fibrous tissues of the tunica propria of the graft was replaced by granulation tissue. Across the anastomoses newly formed capillaries penetrated into the trachea where they caused chondrolysis. This led to a breakdown of the central portion of the prosthesis and stenosis in the airways with terminal respiratory failure. The mucosa of the graft was at some sites covered by metaplastically altered epithelium. Even a reinforcing vascular prosthesis did not prevent the breakdown of the wall of the xenologous graft. An asset for future work is the finding of a well healed suture between the graft and the trachea and evidence that newly formed blood vessels penetrated into the prosthesis. PMID- 7482051 TI - [Artificial retention of the testes]. AB - The author describes artificial retention of the testicle caused by a technical mistake of the surgeon during surgery of an inguinal hernia or hydrocele. The author emphasizes the experience of the surgeon needed during "minor" operations in children. PMID- 7482052 TI - [Thoracoscopy in upper thoracic sympathectomy]. AB - The authors describe their experience with the endoscopic technique of upper thoracic sympathectomy. They evaluate it as an easily performed and safe method which gives great comfort to the patient as well as excellent functional and cosmetic results and involves a minimal burden and ensures rapid convalescence. It enables the surgeon to obtain a perfect orientation in the surgical field as well as safe and perfect preparation of the thoracic portion of the sympathetic nerve. The low cost is also important. PMID- 7482053 TI - [A technic for laparoscopic gastrostomy]. AB - The authors describe the technique of laparoscopic gastrostomy and laparoscopic assisted gastrostomy. It is an alternative method for patients, when PEG (percutaneous endoscopic gastronomy) or other more physiologic way of food administration is not possible to perform. PMID- 7482054 TI - [The T-plate in unstable pertrochanteric fractures of the femur]. AB - The author analyzed and evaluated the therapeutic results of unstable pertrochanteric fractures of the femur treated by the method of a T-shaped splint in 14 patients aged 35-65 years. In 78% during treatment redislocation into the varus position occurred. The mean loss of correction was 36 degrees, despite the fact that a rest regime with a lateral spike splint lasted on average for 32.4 days. In 57% a secondary operation with the necessity of respongioplasty was performed, in two cases final corrective revalgization osteotomy had to be performed. The use of a T-shaped splint in the above indication is controversial. PMID- 7482055 TI - [Surgical problems in acute acalculous cholecystitis]. AB - The authors report on the prevalence of acute cholecystitis in the absence of calculi with special attention paid to the development of this disease as a complication of prolonged parenteral nutrition. In 1990-1993 at the Surgical Clinic in Plzen 125 cases of acute cholecystitis were operated, incl. 14 acalculous ones (11.2%). These 14 patients comprised 5 (28.6%) who were on parenteral nutrition for a prolonged period. The authors analyze the causes of the development of acute acalculous cholecystitis. They deal with the problem of diagnosis where in severe postoperative and posttraumatic cases ultrasonography proved useful. PMID- 7482056 TI - [Pancreatic head resection with preservation of the duodenum--the optimal surgical procedure in chronic pancreatitis]. AB - Surgical procedure appears as an optimal curative method for the treatment of advanced stages of all 4 types of chronic pancreatitis. Progress in diagnostic methods and defining of classification with more precision in last years led us to the idea to optimalize also surgical procedure for the treatment of chronic pancreatitis. Since 1985 we have been performing duodenum preserving pancreas head resection followed by pancreatogastroanastomosis in symptomatic form of chronic pancreatitis. Checking up of 28 patients operated on by this procedure confirm best results of all used methods. With regard to the character of the disease we are not able to improve external secretory function of the pancreas by this operation. External secretory insufficiency has to be substituted permanently by medicaments. Substantial contribution of this operation is in improvement of the quality of the life of the people suffering from chronic pancreatitis. PMID- 7482057 TI - [Pseudomembranous colitis. Personal experience]. AB - The author summarizes experience with the treatment of five patients with pseudomembranous colitis. The diagnosis of this disease was outdated and only in the last two cases it was possible to use the latex agglutination test to assess the presence of Clostridium difficile toxins. The patients were admitted to the clinic in a serious septic toxic condition and thus it proved possible to cure conservatively (using Vancomycin) only one female patient. Four patients were operated in a critical condition (subtotal colectomy with ileostomy and a mucous fistula of the rectosigmoid), two died from progressing sepsis (shock lung). The authors emphasize the necessity of early and accurate diagnosis, incl. assessment of the toxin titre. With regard to the fact that the method is demanding from the economic aspect, the authors propose an economical algorithm of examination and treatment of patients who suffer during prolonged hospitalization from repeated diarrhoea, and the use of broad spectrum antibiotics. PMID- 7482058 TI - [Preoperative drainage of the biliary tract in patients with obstructive jaundice]. AB - The authors present their experience with preoperative external-internal percutaneous transhepatic drainage of the biliary pathways (PTD) in 143 patients. From this group in the final stage 53 patients were operated. The authors discuss the contemporary position and importance of preoperative PTD and compare the advantages of external-internal PTD and external drainage of the biliary pathways. A rapidly performed and correctly indicated external-internal PTD reduces effectively within several days the patient's bilirubin level, the bile passes into the duodenum, dehydration of the organism does not occur and vitamin K absorption is not impaired. All this has an impact on the success of surgery. Moreover, the external-internal drain modified in a suitable way surgical tactics and ensured effective drainage of the biliary pathways during the early postoperative period. If radical surgery is impossible, it is feasible as a suitable alternative procedure of palliative surgery to insert by the percutaneous route an expansible metal stent or leave the external-internal PTD drain in situ for a prolonged period. The authors discuss the criteria for indication, analyze the main problems of this procedure and submit their results. PMID- 7482059 TI - [A method of using intraluminal splints in the treatment and prevention of recurrent ileus of the small intestine]. AB - The authors present a group of 23 patients where splints were used in the loops of the small intestine. The group is divided into two sub-groups. The first one comprises patients where the splint was inserted because of relapsing ileus of the small intestine after previous surgical operations. The second sub-group comprises patients where this operation was performed either preventively or as part of a primary operation or as part of a reoperation called for by complications immediately preceding operation. The internal splint is inserted via jejunostomy and through the loops of the small intestine it is guided by means of a sutured Foley catheter which after proper insertion of the splint is left in the caecum. The authors emphasize the importance of effective decompression of the small intestine during the early postoperative period which is made possible by suction of the ileous contents through lateral openings of the inserted probe. After three years' work, using this method, the authors confirm that it gives good results. PMID- 7482060 TI - [Iatrogenic pneumothorax]. AB - The authors present a group of 22 patients with iatrogenic pneumothorax. They analyze the basic causes of development of iatrogenic pneumothorax, such as cannulation of the subclavicular vein thoracic puncture. They give an account of their experience with the therapeutic procedure which involves the insertion of a classical thoracic drain and several days of active exhausting. In the conclusion they discuss the possibility to use a new method-videothoracoscopy-in some indicated cases. PMID- 7482062 TI - [Treatment of diffuse connective tissue diseases in the past and up to the present]. PMID- 7482061 TI - [Advantages of continuous resorbable mattress sutures in closing extensive laparotomies]. AB - In a group of 170 patients the mean time taken to close the abdominal cavity was 10 minutes, suppuration and fistulation resp. was observed in 2% of patients after 6 weeks. In 1.1% dehiscence occurred and in 3.5% hernia in the scar. All this applies to patients with severe malignity and hypoproteinaemia. Smead Jones technique of a continuous absorbable suture without knots is not only quick, safe, effective, cheap but also reliable in risk patients and obese patients with impaired healing, but only when its basic principles are respected: reliable anchoring and termination in fasciae, regularly alternating distances of stitches. PMID- 7482063 TI - [Methotrexate and salazosulfapyridine in the long-term treatment of rheumatoid arthritis]. AB - Long term treatment of 190 cases of rheumatoid arthritis with either methotrexate or salazosulfapyridine was analyzed for their efficacy and adverse effects. Both treatment groups showed improvement of clinical symptoms, erythrocyte sedimentation rate, and CRP after 1 month of treatment, while RAHA titers decreased significantly after several months. Erythrocyte sedimentation rate and CRP of salazo-sulfapyridine group, once improved, deteriorated again after 12 months of treatment, while methotrexate group showed sustained improvement for 48 months. Radiologic progression estimated according to JF Fries was significantly less in MTX group than in SASP group. Life table analysis showed that the overall probability of continuing methotrexate or salazosulfapyridine at 4 years was 63% and 55%, respectively. The main reason of treatment termination in methotrexate group was adverse effects, while that in salazo-sulfapyridine group was inefficacy. PMID- 7482065 TI - [A case of rheumatoid arthritis with malignant lymphoma taking methotrexate]. AB - We describe one case with rheumatoid arthritis who developed non-Hodgkin's lymphoma during treatment with low dose weekly methotrexate. A 73 year-old man had seropositive RA since 1974. He had been treated with several medications, including nonsteroidal antiinflammatory drugs, gold sodium thiomalate (from January, 1987), and bucillamine (from January, 1988). He presented to this hospital in April 1988, at a time when his rheumatoid arthritis worsened. Methotrexate was administered at a weekly dose of 7.5 mg orally, together with a daily dose of 5 mg of prednisone. He had had no joint-related pain and no side effects until December 1991 (total dose 1290 mg) when severe abdominal pain was started abruptly. The chest X-ray showed an abdominal free air and a diagnosis of acute panperitonitis was made. An emergency operation was carried out. There was a soft-tissue mass in the terminal of ileum which was ruptured with massive ascites. Histologic examination of the mass revealed a diffuse large cell lymphoma. The oncogenic potential of MTX and rheumatoid arthritis is reviewed. PMID- 7482064 TI - [IgG rheumatoid factor in rheumatoid arthritis with interstitial lung disease]. AB - Case of rheumatoid arthritis (RA) with interstitial lung disease (ILD) have been reported to show increased titers of rheumatoid factor (RF) in serum; however the pathogenic role of this substance in the lung is still obscure. The aims of this study were to estimate IgGRF in the bronchoalveolar lavage fluid (BALF) in RA and to investigate its possible roles in the ILD of RA. Two step bronchoalveolar lavage in 20 RA patients (9 males, 11 females) was performed in conjunction with high resolution chest CT. IgGRF was measured by enzyme-linked immunosorbent assay and its molecular size in BALF was evaluated by high performance liquid chromatography. IgGRF bound to BALF cells was eluted by sonication of the cells. RA patients with ILD had significantly elevated levels of IgGRF in serum as well as in BALF. The IgGRF in BALF was monomeric in 2 of 3 RA patients with ILD. The activity of IgGRF bound to BALF cells was significantly higher in RA with ILD than in control patients. These findings suggest that monomeric IgGRF in the lungs of RA patients could bind to the cells with Fc receptors. PMID- 7482067 TI - [Systemic lupus erythematosus and antiphospholipid antibody syndrome in a patient with congenital ichthyosiform erythroderma]. AB - A 23-year-old woman, who had nonbullous congenital ichthyosiform erythroderma since her childhood, was diagnosed as nephrotic syndrome caused by systemic lupus erythematosus (SLE). She was pregnant but experienced fetal loss at the age of 25. Although 10 mg/day of oral prednisolone was administered, low levels of serum complement, proteinuria, thrombocytopenia (6.0 x 10(4)/mm3) and biological false positive for STS continued. When she was 27 years old, right hemichorea developed. She was admitted to our hospital at the age of 28 because of low levels of serum complement, high titers of anti ds-DNA antibody, profuse proteinuria, gingival bleeding and thrombocytopenia (1.5 x 10(4)/mm3). The nephrotic syndrome gradually improved after 1 g/day of methylprednisolone for 2 days and the oral prednisolone dosage was then increased up to 40 mg/day, and was tapered to 10 mg/day. Epileptic attack (minor seizure) occurred at the age of 29. Continuous low levels of serum complement and high titers of anti ds-DNA antibody were improved by adding 50 mg/day of cyclophosphamide. However, high levels of beta 2 GPI dependent anticardiolipin antibody and lupus anticoagulant activity were observed throughout the study. Our patient was a very rare case of congenital ichthyosis with typical antiphospholipid antibody syndrome and SLE. A few cases of acquired ichthyosis associated with SLE has been reported, and ichthyosis developed only in active stage of SLE. However, our patient's ichthyosiform lesions were not changed throughout the course. PMID- 7482066 TI - [A case of p-ANCA positive necrotizing vasculitis associated with Graves' disease]. AB - A 22-year-old female who had been suffering from Graves' disease developed p-ANCA positive necrotizing vasculitis. Her Graves' disease has been treated with propylthiouracil (PTI) for 6 years, which failed to improve serum TSH. She developed transient and recurrent polyarthralgia 4 years before. Because of microscopic hematuria and polyarthralgia, She visited our hospital last year. She had hypergammaglobulinemia and microhematuria at that time. But no autoantibody was noted except anti-microsomal antibody at that time. Skin biopsy from her finger subdermal nodule revealed necrotizing vasculitis of small vessels. Serum p ANCA was positive. She was admitted under the diagnosis of p-ANCA positive necrotizing vasculitis. Renal biopsy revealed mesangium proliferative glomerulonephritis. Since she developed telescoped sediments after admission, predonisolone (PSL) 50 mg daily was started. Clinical and laboratory findings improved quite well. PSL was gradually tapered after 6 weeks. She is being maintained in good condition on tapering doses of both PSL and PTI. The coincidence of Graves' disease and vasculitis is rare. This case may provide some important immunological insights into the study of pathogenesis of necrotizing vasculitis and Graves' disease. PMID- 7482068 TI - [An autopsy case of segmental mediolytic arteritis (SMA) accompanied with microscopic polyarteritis nodosa]. AB - Segmental Mediolytic Arteritis (SMA) is a very rare arterial lesion which is limited in adults to the involvement of the intra-abdominal muscular arteries. The pathology is characterized by segmental disruption of the arterial media which leads segmental mediolysis, with subsequent dissecting aneurysm or rupture. A 73-year-old man was admitted to a hospital because of high fever, general fatigue and weight loss. These symptoms were resistant to antibiotic therapy, and soon after, renal insufficiency developed. Three months after the onset of symptoms, he died suddenly of hemorrhagic shock. Autopsy revealed rupture of the splenic artery and systemic necrotizing arteritis in the small-sized arteries of liver, pancreas and kidneys, as well as in the connective tissues around the adrenal glands. Histopathology of the splenic artery was consistent with SMA, and that of the systemic vascular lesions, with microscopic polyarteritis nodosa. Focal glomerular lesions characteristic of crescentic and/or granulomatous glomerulonephritis were present. A developmental mechanism for SMA is discussed with respect to this case together with a review of previous reports of this disease. PMID- 7482069 TI - [Pneumocystis carinii pneumonia associated with low dose methotrexate treatment for malignant rheumatoid arthritis]. AB - Low dose pulse methotrexate (MTX) has become a widely used therapy for rheumatoid arthritis (RA) because of its good response rate profile. With the increased use of MTX, reports of opportunistic infections associated with MTX therapy have appeared. Fourteen cases of pneumocystis carinii (PC) pneumonia in patients receiving low dose MTX have been previously reported. Yet, no case of PC pneumonia associated with low dose MTX has so far been reported in Japan. We report the first case in Japan of PC pneumonia occurring in a patient with rheumatoid vasculitis who was receiving low dose MTX. A 70-year old woman with 13 year history of RA presented with 3-day history of rapidly aggravating dyspnea, dry cough and fever. She had been receiving MTX 7.5 mg/week for 2.5 months because of her vasculitis symptoms. She had also been receiving prednisolone 7.5 mg/day which had been successfully tapered from an initial dose of 15 mg/day. At the time of her presentation with respiratory symptoms, all of her vasculitis symptoms had been alleviated. A chest radiograph revealed diffuse interstitial shadowing bilaterally and bilateral hilar and right lower lung field infiltrates. Her arterial blood gas showed severe hypoxemia (PaO2 27.7 torr). Polymerase chain reaction assay of bronchoalveolar lavage fluid showed PC. Although the patient required ventilatory support for 9 days, she was successfully treated with trimethoprime-sulphamethoxazole and methylprednisolone pulse therapy. Eight months later, the patient was well with no evidence of vasculitis or respiratory symptoms. PMID- 7482070 TI - [Retrovirus and autoimmune diseases]. PMID- 7482071 TI - [Apoptosis in autoimmune diseases]. PMID- 7482072 TI - Reference Centre to address STDs. PMID- 7482074 TI - Towards health-promoting schools in South Africa. PMID- 7482073 TI - Latest figures on HIV pregnancies. PMID- 7482075 TI - The prevalence and prevention of neural tube defects in Cape Town. PMID- 7482076 TI - Can we still justify the use of low-dose thiazide diuretics as first-line treatment for black South African hypertensives? PMID- 7482077 TI - Who promotes primary health care and why? PMID- 7482078 TI - The introduction of surfactant replacement therapy into South Africa. AB - Within the context of limited financial and physical resources in South Africa, academic neonatologists have established strict criteria for ventilation of neonates with hyaline membrane disease (HMD). In the private sector neonatal care is less structured. Following the introduction of the artificial surfactant (Survanta) in November 1991 it was considered important to monitor its use in the public and private sectors. In collaboration with the marketing company a data sheet containing demographic information and clinical details was drawn up to be completed in each case where Survanta was administered to babies with HMD. Data from 155 babies treated at 10 hospitals were included in the final analysis (70 babies from 4 State-funded academic hospitals and 85 from 6 privately funded hospitals). Within the group of private hospitals there were some which treated large numbers of babies weighing > 2,000 g, while in a few others there was a relative excess of babies weighing < 1,000 g. There was a higher incidence of patent ductus arteriosus and intraventricular haemorrhage, and a higher mortality rate at the academic hospitals. Poor outcome at these institutions may largely have been due to poor antenatal care. PMID- 7482079 TI - The selection of infants for surfactant replacement therapy under conditions of limited financial resources. AB - The cost of surfactant replacement therapy (SRT) will restrict its use under conditions of limited health resources. Before the local advent of SRT, infants ventilated for hyaline membrane disease (HMD) had an overall survival rate of 87% and an incidence of bronchopulmonary dysplasia of 6.4%. This, together with the cost of SRT, prompted a study to identify those infants who would benefit the most from SRT. Twenty-two infants assessed as having severe HMD were randomised to receive SRT at 3 - 4 hours (9) or at 6 - 8 hours (13) after birth. Two infants (15%) in the latter group did not require SRT. The outcome of these two groups was the same. Of 56 infants assessed as having moderate HMD, only 24 (43%) qualified for SRT from 6 hours of age. The outcome of the SRT and non-SRT infants was comparable. The group of infants with moderate HMD had a significantly better outcome than those with severe HMD. A limited period of observation to assess the severity of illness did not compromise outcome in this group of 78 infants with moderate to severe HMD. PMID- 7482080 TI - The late administration of surfactant. AB - Current recommendations for surfactant replacement therapy (SRT) in the treatment of hyaline membrane disease (HMD) are to administer the drug as soon as possible after starting ventilation in order to prevent ventilator lung damage. We present a review of 18 infants (gestational age 32.4 +/- 1.9 weeks and birth weight 1,795 +/- 427 g) who received the initial dose of SRT after they were 12 hours old. Fourteen infants were assessed as having HMD and 4 as having congenital pneumonia. Overall there was a significant and sustained improvement in oxygenation as measured by arterial/alveolar oxygen ratios. The outcome of these infants was good, with a duration of ventilation of 7.9 +/- 4.3 days and a duration of hospitalisation of 26.2 +/- 12.6 days. No infant developed bronchopulmonary dysplasia. Of particular interest is that 3 infants weighing > 2,400 g with congenital pneumonia responded to a single delayed dose of SRT. Late SRT is effective and there may be a place for SRT in the treatment of conditions other than HMD. PMID- 7482081 TI - The cost and effectiveness of surfactant replacement therapy at Johannesburg Hospital, November 1991-December 1992. AB - OBJECTIVE: To assess the impact of surfactant replacement therapy (SRT) on the outcome of hyaline membrane disease (HMD) and to assess the cost implications of a policy of selective administration of artificial surfactant. DESIGN: The short term outcome of 103 newborns ventilated for HMD (61 selected for SRT according to initial and/or ongoing oxygen requirements) was compared with that of a historical control group of 173 infants ventilated for HMD before the introduction of SRT. MAIN OUTCOME MEASURES: Mortality and morbidity of HMD including death, bronchopulmonary dysplasia, pneumothorax, pulmonary haemorrhage, patent ductus arteriosus and intraventricular haemorrhage. RESULTS: There were significant demographic differences between the treatment and control groups (black patients 74% v. 28%, P < 0.0001; unbooked mothers 72% v. 15%, P < 0.0001) as well as evidence of more severe lung disease in the treatment group (pressor support 44% v. 27%, P < 0.005; and paralysis during ventilation 38% v. 25%, P < 0.005). Pneumothorax was reduced in the SRT group (7% v. 17%, P < 0.01). There were no significant differences between the two groups in the incidence of BPD or mortality. The use of SRT added to the total cost of treating a patient ventilated for HMD. CONCLUSION: The selective use of SRT had the effect of converting severe disease into moderate disease rather than achieving maximal benefit in all cases of HMD through routine use of the product. A policy of restricting use may result in cost savings where resources are limited. PMID- 7482082 TI - The outcome at 12 months of very-low-birth-weight infants ventilated at Tygerberg Hospital. AB - OBJECTIVE: To determine the outcome at 1 year of age of a group of very-low-birth weight (VLBW) infants, from urban and rural communities, ventilated at Tygerberg Hospital, W. Cape. STUDY DESIGN: Prospective descriptive study in which the prevalence of bronchopulmonary dysplasia (BPD), sensorineural deafness, intraventricular haemorrhage (IVH), retinopathy of prematurity (ROP) and abnormal motor developmental outcomes were determined in 153 ventilated VLBW infants from rural and urban areas. Of these, 69% were from lower socio-economic backgrounds. MAIN OUTCOME MEASURES: Attrition rates for rural and urban babies, BPD, ROP, IVH and abnormal motor development. STUDY POPULATION AND SETTING: All ventilated VLBW infants discharged from the neonatal intensive care unit at Tygerberg Hospital over a 1-year period were followed up at 3-monthly intervals for 12 months. RESULTS: BPD was diagnosed in 19% of the babies, with significantly more babies with birth weights under 1,000 g and gestational ages under 28 weeks having BPD. Of the babies with BPD, 25% had abnormal motor development at 1 year of age. Seven per cent of the babies had grade 3 or 4 ROP and 2.6% had sensorineural hearing loss. One hundred and seventeen (79%) of the infants attended the follow up clinic until 12 months of age (corrected for prematurity). There were no significant differences in the number of babies followed up from rural or urban areas. Fourteen (11.9%) of the babies had abnormal motor development. A disturbing finding was that so many babies had spastic quadriplegia (8; 57%) versus diplegia (6; 43%). The incidence of abnormal motor development in the babies from the rural areas and in those with birth weights under 1,000 g from rural areas was high--a further cause for concern. CONCLUSION: The prevalence of the major complications associated with ventilated VLBW infants correlated well with those reported for similar infants from First-World countries. The poor motor developmental outcome of the babies from rural areas with birth weights under 1,000 g and high attrition rates for infants with serious complications such as BPD, IVH and ROP are distressing. PMID- 7482083 TI - Paediatric diarrhoea--rehydration therapy revisited. AB - Acute infective diarrhoea remains one of the most common causes of morbidity and mortality in children under 5 years of age. This paper reports on a 7-year experience of management of paediatric diarrhoea at King Edward VIII Hospital, Durban, in which an inpatient case-fatality rate of nearly 25% was reduced to less than 7%, and the admission rate was reduced by 60% by a cumulative effect of the following measures: one of four paediatric wards was converted into a diarrhoea ward; improved attention to protocol resulted in a rapid reduction in the inpatient case-fatality rate, but further improvement resulted from a strong commitment to an efficient outpatient oral rehydration protocol to reduce the pressure on inpatient beds, as well as a simplified approach to fluid therapy; and a formula was developed based on units of 5 ml/kg/h, and applicable to both oral and intravenous routes. The most important assessment of dehydrated patients is determination of a need for resuscitation. In less severely ill patients it is not necessary to calculate rehydration fluid requirements by a 'percent dehydration' formula. PMID- 7482084 TI - TB or not TB? An evaluation of children with an incorrect initial diagnosis of pulmonary tuberculosis. AB - OBJECT: The aim of the study was to identify diagnoses that are confused with pulmonary tuberculosis in children. DESIGN: Prospective, investigative clinical study. SETTING: Tertiary care teaching hospital and an urban tuberculosis clinic in an area with a very high incidence of pulmonary tuberculosis (> 800 new cases/100,000/year). PATIENTS: Children suspected of having tuberculosis, children followed up for pulmonary infiltrates with eosinophilia and children with congenital pulmonary anomalies were investigated. INTERVENTION(s). None. OUTCOME MEASURE: Pulmonary tuberculosis was diagnosed using modified World Health Organisation criteria and the diagnoses of those children not suffering from pulmonary tuberculosis were analysed. RESULTS: Of the 354 children initially suspected of suffering from tuberculosis 71 (20%) were found to be suffering from other pulmonary disease, viz. pneumonia or bronchopneumonia (29%), bronchopneumonia with wheezing (18%), and asthma with lobar or segmental collapse (12%). Of 14 children suffering from pulmonary infiltrates with peripheral eosinophilia 6 (43%) were initially incorrectly diagnosed and treated for tuberculosis. Of 54 children with congenital pulmonary anomalies, 8 (15%) were treated for tuberculosis before the correct diagnosis was made. Congenital anomalies most often confused with tuberculosis were unilateral lung hypoplasia, bronchogenic cyst and tracheal bronchus with an anomalous lobe. CONCLUSIONS: The criteria for diagnosing tuberculosis in children is complicated in areas with a high incidence of tuberculosis and poor socio-economic circumstances where many children presenting with conditions other than tuberculosis will be in contact with an adult case of pulmonary tuberculosis. The commonest conditions confused with tuberculosis are pneumonia, bronchopneumonia and asthma. Pulmonary infiltrates with peripheral eosinophilia and congenital lung abnormalities should be considered especially if the children have an atypical clinical picture or do not respond to tuberculosis treatment. PMID- 7482085 TI - Subclinical pertussis in incompletely vaccinated and unvaccinated infants. AB - Incidental to a phase II study of acellular and whole-cell pertussis vaccines involving 342 infants who were clinically observed from birth until the age of 9 months, subclinical pertussis was retrospectively diagnosed in 10 infants on the basis of serological evidence. IgG and IgA to filamentous haemagglutinin (FHA), pertussis toxin (PT) and agglutinogens 2 and 3 (AGG2, 3) were assayed by enzyme linked immunosorbent assay (ELISA) in serum obtained at birth and at 2, 4, 6 and 9 months of age. All 10 infants had > or = 4-fold rises in at least two different pertussis IgG antibodies. Nine of the 10 infants had > or = 4-fold increases in all three IgG antibodies measured. One infant had > or = 4-fold increases in IgG FHA and IgG-AGG2,3 but not IgG-PT. Seven infants had raised IgA antibodies to PT and FHA and 4 infants had raised IgA antibodies to AGG2,3. Subclinical infection provoked differing degrees of antibody production in response to multiple antigens. Subclinical infection was detected in both unvaccinated infants (4) and in infants who had been vaccinated from 2 months of age with either acellular (4) or whole-cell vaccines (2). Subjects were 8 months of age or younger and only 1 had completed primary vaccination. Other infections of infancy were commonly detected; 4 infants had upper respiratory disease about the time of subclinical pertussis. None had a household member with symptomatic pertussis. Likelihood of subclinical infection was related to significantly lower levels of maternally acquired pertussis IgG-AGG2,3 antibodies but not associated with infants' nutritional status. PMID- 7482086 TI - Rickets in black children beyond infancy in Natal. AB - OBJECTIVE: To determine the clinical spectrum of rickets among black children admitted to King Edward VIII Hospital, Durban. DESIGN: Prospective study of black children with rickets beyond infancy. SETTING: Hospital-based population; King Edward VIII Hospital, Durban. PARTICIPANTS: A total of 37 patients, aged 1-12 years, were recruited over a 3-year period. None had been on vitamin D or calcium supplementation prior to investigation. OUTCOME MEASURES: Rickets was diagnosed clinically, radiologically and biochemically (by a raised alkaline phosphatase value of > 350 IU). Gastro-intestinal, hepatic and renal glomerular causes were excluded in all patients using standard clinical and laboratory criteria. RESULTS: Twenty-three patients were diagnosed as having privational rickets. Nine had 25-hydroxyvitamin D (25-OHD) levels of < 10 ng/ml while 14 had levels within the normal range and were suspected of having dietary calcium deficiency. Ten had a phosphopenic variety of rickets; the remaining 4 had healing or healed rickets on the basis of radiological assessment and normal biochemical values. Pain together with difficulty in walking and bowing of the lower limbs were the main reasons for presentation. The main clinical findings were thickened wrists and ankles and rickety rosary (100%), stunting (85%), anterior bowing of lower limbs (70%) and genu valgum (65%). The calcium and vitamin D deficiency group showed a much better clinical, biochemical and radiological response to therapy than the phosphopenic group on follow-up (18 patients). CONCLUSION: This is the first substantial report on rickets in the older child in Natal, which extends the findings from Transvaal, thereby establishing a recognisable pattern of rickets beyond infancy in South Africa. It draws attention to the common clinical presentations which may alert health professionals to the presence of this problem. This report demonstrates that the two commonest types are privational rickets (due to calcium and/or vitamin D deficiency) and phosphopenic rickets. PMID- 7482087 TI - Extradural haematomas in children. AB - OBJECTIVE: To determine the cause, clinical presentation, radiological and computed tomography (CT) findings, anatomical position and post-surgical outcome of traumatic intracranial, extradural haemorrhage in children. DESIGN: Retrospective study, 1979-1994. SETTING: Neurosurgical referrals from the Trauma Unit at Red Cross War Memorial Children's Hospital, Cape Town. PARTICIPANTS: Forty-four children below the age of 12 years who had extradural haematomas surgically removed. RESULTS: Twenty-one extradural haematomas resulted from falls, 12 from motor vehicle accidents, and 6 from assaults; 1 was of unknown cause. Nineteen patients presented with persistent drowsiness (14/15 on the Glasgow Coma Scale), 17 had headache and vomiting, and 13 gave a history of initial loss of consciousness. Seventeen pre-operative neurological examinations were normal, 11 showed anisocoria, 10 hemiplegia, 4 ataxia, 1 nystagmus and 1 a full fontanelle. Cases of posterior fossa haematoma presented with headache and ataxia. Skull fractures were visible on radiographs in 26; computed tomographic diagnosis was made in all but 1. The site of extradural haematoma was parieto occipital in 24, posterior fossa in 11, frontal in 6, and temporal in 3. Subdural haematomas occurred in 4 cases. Surgical treatment consisted of 41 craniotomies/craniectomies and 3 burr-holes. Postoperatively 36 patients were neurologically normal, 6 had neurological deficits, and 2 died. CONCLUSION: Intracranial extradural haemorrhage in children presents in an atypical manner when compared with adults. An initial loss of consciousness followed by a lucid interval and a progressive deteriorating level of consciousness is the exception rather than the rule. Timeous neurosurgical treatment resulted in an excellent outcome in 36 of the 44 children. PMID- 7482089 TI - Ending physical punishment of children in South Africa. PMID- 7482088 TI - Diagnosis and treatment of ear disease among children in the Ellisras district. An outreach programme. AB - Audiologists and ENT registrars examined 2,036 children aged 10 years or younger by means of pure-tone audiometry, tympanometry and otoscopic examinations. Twenty per cent of these children had ear pathology and 7.5% had impaired hearing. Forty three pus swabs taken from patients with suppurative otitis media most often cultured Proteus mirabilis and Pseudomonas species. The prevalence of otitis media with effusion was twice as high in white children as in black children, resulting in a much higher prevalence of hearing loss in the first group. The black children had 7 times more dry perforations of the tympanic membrane and 11 times more wet perforations. A hundred and thirty-six patients were treated by the team and 137 were referred for further treatment. Sixteen operations were performed at a later stage. Further programmes are needed to diagnose and treat ear disease in rural communities. PMID- 7482090 TI - Caterpillar in the eye--an ophthalmic emergency! PMID- 7482091 TI - Transformation of the image of the general practitioner in the new South Africa. PMID- 7482092 TI - Abortion--who should decide? PMID- 7482093 TI - Co-operation between traditional healers and medical personnel. PMID- 7482094 TI - Malignant hyperthermia--still around. PMID- 7482095 TI - Infantile onset of HTLV-I-associated myelopathy (HAM/TSP) PMID- 7482096 TI - Helicobacter pylori positivity in asymptomatic children of different ethnic groups living in the same country. PMID- 7482098 TI - Breast-milk substitutes. PMID- 7482097 TI - Breast-milk substitutes. PMID- 7482099 TI - Breast-milk substitutes. PMID- 7482100 TI - Breast-milk substitutes. PMID- 7482101 TI - Control of epidemic early syphilis: the results of an intervention campaign using social networks. AB - BACKGROUND: During an epidemic of early syphilis, social networks were used for an intervention campaign. GOAL OF THIS STUDY: To characterize the epidemic and describe the yield of new cases from index-case interviews. METHODS: Analyses of morbidity data collected by the Montgomery County, Alabama, sexually transmitted disease program determined the course of the epidemic and characterized the new case yields from social networks identified via index-case interviews (partner notification investigations) and interviews with sex partners and their associates (cluster investigations). Results and costs were compared to a noncampaign period. RESULTS: The number of reported syphilis cases nearly doubled from 1990 to 1991 (201 to 348 per 100,000 residents). During the 21-week campaign, 373 case-patients had partner notification/cluster investigations; 113 (11%) of 984 sex partners and 41 (3%) of 1,146 high-risk associates (persons identified during cluster investigations) had syphilis. No subgroup of case patients for which the partner notification/cluster investigation yielded more infected persons than other subgroups was identified. The cost per case detected was more than twice that during a noncampaign period ($1,627 vs. $771). CONCLUSION: Partner notification investigations yielded more infected persons than cluster investigations. Further evaluation is needed to determine the role of intense partner notification/cluster investigators' efforts in the control of epidemic syphilis. PMID- 7482102 TI - Herpes simplex virus-2 may increase susceptibility of the sexual transmission of hepatitis C. AB - OBJECTIVES: Antibodies against herpes simplex viruses-1 and -2, cytomegalovirus, and syphilis were determined in six heterosexual couples with strong indications of having sexually transmitted hepatitis C virus infection and in 17 other heterosexual couples in which one partner was hepatitis C virus viremic (source partner), but the other had remained hepatitis C virus uninfected (exposed partner). STUDY DESIGN: Antibody testing was done with an enzyme-linked immunosorbent assay. Anti-herpes simplex virus 2 and anti-hepatitis C virus findings were further confirmed by immunoblotting. Hepatitis C virus RNA was determined by polymerase chain reaction and genotyped with type-specific primers. RESULTS: Five of six anti-hepatitis C virus-positive exposed heterosexual partners without parenteral risk factors, compared with three of 17 anti hepatitis C virus-negative exposed partners, had antibodies to herpes simplex virus-2. On the other hand, no statistically significant difference was found regarding the frequency of herpes simplex virus-2 seropositivity when source partners in the anti-hepatitis C virus concordant and discordant couples were compared. The presence of antibodies to herpes simplex virus-1, cytomegalovirus, and syphilis did not significantly differ between source or exposed partners in anti-hepatitis C virus concordant and discordant couples, respectively. No predominance of any one hepatitis C virus genotype or liver morphology in couples concordant compared with discordant for anti-hepatitis C virus was found. CONCLUSIONS: The findings support the role of herpes simplex virus-2 in the heterosexual transmission of hepatitis C virus infections, and more specifically an increase in susceptibility to hepatitis C virus infections in exposed heterosexual partners with antibodies to herpes simplex virus-2. PMID- 7482103 TI - Incision and drainage versus aspiration of fluctuant buboes in the emergency department during an epidemic of chancroid. AB - GOAL OF THIS STUDY: To compare outcomes of incision and drainage with packing to needle aspiration of purulent buboes in patients with presumed chancroid. The safety and efficacy of incision and drainage were compared with that of needle aspiration. STUDY DESIGN AND METHODS: The study was a prospective randomized, nonblinded clinical trial carried out from April 1992, to January 1994 in an inner city emergency department. A consecutive sample of 27 adults with fluctuant inguinal buboes presumed or proven to be chancroid or lymphogranuloma venereum were eligible for inclusion. Patients were excluded if they were believed to have buboes secondary to disease other than chancroid. After informed consent was obtained, patients had buboes drained by needle aspiration or by incision and drainage with packing according to a random numbers table. RESULTS: Twenty-seven patients were included in the study (22 men and 5 women), with 12 randomized to incision and drainage and 15 to aspiration. The mean age was 35.7 +/- 13 years. Genital ulcer or bubo pus cultures for Haemophilus ducreyi were positive in seven patients, negative in 15 patients, and not done in five patients. Follow-up was obtained for 23 (85%) patients, 11 of whom had incision and drainage and 12 of whom had aspirations. No adverse effects were reported in either group. CONCLUSION: Incision and drainage is an effective method for treating fluctuant buboes and may be preferable to traditional needle aspiration considering the frequency of required re-aspirations in the study patients. Limitations of this study include lack of complete laboratory testing and lack of follow-up of all patients. PMID- 7482104 TI - Screening for Chlamydia trachomatis infection in pregnant women in Martinique. AB - GOAL OF THIS STUDY: To determine the prevalence of Chlamydia trachomatis urogenital infection and to identify behavioral, demographic, and clinical factors associated with the infection in pregnant women in Martinique. STUDY DESIGN: One-thousand-four-hundred-eleven patients 15-39 years old, at 10-16 weeks of gestation and attending the prenatal clinic at Lamentin Hospital, were tested for Chlamydia trachomatis infection of the cervix and urethra using tissue culture. RESULTS: Chlamydia trachomatis was isolated from 375 (26.7%) women; 34% of them were positive in the cervix and urethra, 58% in the cervix only, and 8% in the urethra only. Factors found by multivariate analysis to be significantly associated with chlamydial infection were age less than 25 years, first intercourse at less than 18 years old, previous induced abortions, mucopurulent cervicitis, and repeated candidiasis. CONCLUSIONS: None of the factors associated with chlamydial infection was sensitive enough to permit efficient selective screening. It is cost effective to recommend a routine screening for chlamydial infection together with an educational program. PMID- 7482105 TI - Comparative study of fluconazole and clotrimazole for the treatment of vulvovaginal candidiasis. AB - BACKGROUND: The introduction of the triazole antifungal agent fluconazole, which can be used via an oral route and single dose, has had a significant impact on patient compliance. GOAL OF THIS STUDY: To compare the efficacy and safety of a single oral dose of 150 mg fluconazole with that of a 100-mg clotrimazole vaginal suppository twice a day in the morning and at bedtime for 3 days in the treatment of vulvovaginal candidiasis. STUDY DESIGN: A single-blind, randomized controlled trial was conducted at Ramathibodi Hospital between June 1 and September 30, 1993. RESULTS: There were 53 women in the group treated with fluconazole and 50 in the group treated with clotrimazole. There was no significant difference between the two groups regarding clinical characteristics (age and length of follow-up period). Mycological cure rates approximately 1 week after treatment were 79.2% in the fluconazole group and 80.0% in the clotrimazole group. Approximately 4 weeks after treatment, these rates were 60.4% and 66.0%, respectively. The side effects were minimal and did not warrant any treatment. The differences in the results were not statistically significant. CONCLUSION: We recommend that a single oral dose of 150 mg of fluconazole be given as an alternative method of treating vulvovaginal candidiasis, but the cost is higher than for clotrimazole. PMID- 7482106 TI - STD partner notification and referral in primary level health centers in Nairobi, Kenya. AB - BACKGROUND: Controlling sexually transmitted diseases requires that partners of patients with a sexually transmitted disease be notified and treated. However, many countries in the developing world lack the infrastructure and resources for effective partner referral. GOAL OF THIS STUDY: To provide information on rates of partner referral in primary-level health centers in Kenya, to identify characteristics of patients with sexually transmitted diseases who inform their partners about the need for treatment, and to evaluate the impact of a brief counseling intervention on rates of partner notification. STUDY DESIGN: Two hundred-fifty-four patients presenting for treatment of a sexually transmitted disease were given 5 to 10 minutes of additional counseling on the importance of referring partners for sexually transmitted disease treatment. All patients who returned for follow-up 1 week later were interviewed to determine whether they had notified their sex partners. RESULTS: Sixty-eight percent of patients who returned for follow-up reported they had referred their partners for treatment of a sexually transmitted disease. The highest rates of partner notification occurred among women attending maternal child health/family planning clinics and married men and women attending general outpatient clinics. CONCLUSION: Strengthening and directing counseling toward women in maternal child health/family planning clinics and married men and women in general clinics may be an effective and inexpensive way to increase partner notification in the developing world. PMID- 7482108 TI - Male urethritis with and without discharge: a clinical and microbiological study. AB - BACKGROUND: The definition of male urethritis in the absence of urethral discharge has not been well established. The sensitivity of urethral swabs and first-catch urine is controversial. GOAL OF THIS STUDY: To correlate clinical data (discharge or not), urethral swabs, and first-catch urine examinations with the microorganisms found within the urethra in a cohort of men attending the sexually transmitted disease clinic of Hopital Saint Louis (Paris) for treatment of urethral symptoms with or without discharge. STUDY DESIGN: Two-hundred-seventy three consecutive male patients entered this prospective study between October 1, 1992 and November 30, 1992. Fifty-two patients were excluded because they had been treated with antibiotics in the previous 3 months. All patients were screened for Chlamydia trachomatis, Neisseria gonorrhoeae, Mycoplasma genitalium, Trichomonas vaginalis, Ureaplasma urealyticum, Mycoplasma hominis, and Candida albicans. RESULTS: Two-hundred-nineteen patients were eligible for the study (122 with discharge and 97 with no discharge). The prevalence of microorganisms was as follows: Chlamydia trachomatis in 13%, Neisseria gonorrhoeae in 11%, Ureaplasma urealyticum in 7%, Mycoplasma genitalium in 17%, Trichomonas vaginalis in 1%, and indeterminate pathogens alone in 20%. All major pathogens and Mycoplasma genitalium were more common in patients with discharge. Stratification of results according to the presence of polymorphonuclear leukocytes on the urethral swab and first-catch urine showed a low sensitivity of both tests for Chlamydia trachomatis (29%), Mycoplasma genitalium (50% and 62%), and Ureaplasma urealyticum (33%) in patients with no discharge. CONCLUSION: A specific and sensitive search for Chlamydia trachomatis should be done in every patient with urethral symptoms whether or not the classic symptoms of urethritis are present (discharge, presence of polymorphonuclear leukocytes in the urethra or first catch urine). PMID- 7482107 TI - Spectrum of genital human papillomavirus infection in a female adolescent population. AB - BACKGROUND: Human papillomavirus infection is a sexually transmitted disease associated with cervical dysplasia and carcinoma. GOAL OF THIS STUDY: To determine prevalence rates of cervical human papillomavirus infection compared with other sexually transmitted diseases and risk factors associated with human papillomavirus infection among adolescent women, we evaluated 634 patients attending three urban adolescent clinics. STUDY DESIGN: Patient evaluation included Pap smears; screening for chlamydia, gonorrhea, and trichomoniasis; and testing of cervical swab samples for human papillomavirus DNA. RESULTS: Cervical human papillomavirus was the most common STD in our population (15.6%), followed by infection with Chlamydia trachomatis (11.0%), Neisseria gonorrhoeae (7.1%), and Trichomonas vaginalis (5.4%). The most prevalent human papillomavirus types were 16/18 (7.3%), followed by 31/33/35 (4.7%) and 6/11 (3.5%). When genital warts on exam, low-grade squamous intraepithelial lesions on cytology, or cervical human papillomavirus DNA were considered as indicators of genital human papillomavirus infection, 24% of patients had any manifestation of infection, including 15% with clinically apparent infection (genital warts), 36% with cytologically apparent infection without warts, and 49% with subclinical infection only (cervical human papillomavirus DNA without low-grade squamous intraepithelial lesions or warts). Factors associated with detection of cervical human papillomavirus DNA by multivariate analysis included number of lifetime sexual partners and genital warts on exam. CONCLUSION: Cervical human papillomavirus infection was the most prevalent sexually transmitted disease among an ethnically diverse group of urban adolescent females, with a large proportion of infections neither clinically nor cytologically apparent. The strong association with lifetime sexual partners substantiates that cervical human papillomavirus is acquired predominantly by sexual contact and often soon after the onset of sexual activity. PMID- 7482109 TI - Genital Chlamydia trachomatis infections in Uppsala County, Sweden, 1985-1993: declining rates for how much longer? AB - BACKGROUND: In Uppsala County, between 1985 and 1993, examinations for Chlamydia trachomatis were being performed in a central laboratory. A change in national sexually transmitted disease legislation in 1988 encouraged screening. GOALS OF THIS STUDY: To analyze trends in detection rates of genital chlamydial infections by age, sex, and clinic type, and to assess the influence of the legislation change. STUDY DESIGN: This was an analysis of 119,892 tests, representing 95.4% of all specimens sampled. Eighty-six percent of the samples were cultured, 14% were examined by enzyme immunoassay. RESULTS: Seventy-nine percent of specimens came from women. 7,989 positive samples were identified. Detection rates declined from 107.2 per 1,000 in 1985 to 32.3 in 1993 in women and from 183.3 to 70.7 in men. Positivity rates were highest in sexually transmitted disease and youth clinics and lowest in private practices. Among female youths, rates leveled off in later years, but the rates increased in male youths. The legislation change reduced the probability of a positive test in men but not in women. CONCLUSIONS: Genital chlamydial infection generally declined. However, among youths, an increase has occurred in recent years. Continued screening and the introduction of noninvasive diagnostic methods for males is warranted, particularly in youth clinics. Selective screening may be more cost-effective in other age groups. PMID- 7482111 TI - [The hematologist and the hematology and hemotherapy specialty in general county hospitals]. PMID- 7482110 TI - Acquired immunity to gonorrhea? PMID- 7482112 TI - [Ki-1-positive anaplastic large-cell lymphoma. Clinical significance]. PMID- 7482113 TI - [Anaplastic lymphoma versus Hodgkin's disease. Diagnosis, Controversies]. PMID- 7482114 TI - [Blood donors seropositive for type 1 human immunodeficiency virus (HIV-1). Epidemiologic characteristics and clinical course]. AB - PURPOSES: To evaluate retrospectively the epidemiologic profile and clinical course of the HIV-positive voluntary blood donors in the Sant Pau's Hospital Blood Bank (Barcelona-Spain) between 1986 and 1993. PATIENTS AND METHODS: A total of 119,345 blood donors were studied, and fifty seven such cases were identified. Risk behavior, causes of failure of the pre-donation procedures and clinical stage were analyzed. A follow-up of their infection was performed by the Infectious Disease Unit. RESULTS: The mean age of 57 seropositive donors was 31 +/- 8 years. Prevalence of HIV-1 infection among donors was 0.48 per 1000. Forty four (77%) were male and 13 (23%) female. Forty three (75%) were first time donors while 14 (25%) were repeat donors. The causes for the failure of the self deferral questionnaire were: 42 subjects did not consider that they had engaged in "risk" behavior, seven donors lied in order to know if they were HIV carriers and two cases were driven to donate due to social or family pressure. The distribution of these donors according to risk behaviour was: 30 (53%) heterosexual, 11 (20%) homosexual, 11 (20%) intravenous drug users and five (7%) with no identified risk. It was noted that HIV infection progressed more rapidly to AIDS in HIV - positive homosexual donors than in heterosexual subjects (p < 0.05). CONCLUSIONS: Heterosexual donors who maintain sexual contacts with different partners without prophylactic measures for HIV infection currently represent the highest risk group for transfusion-related HIV infection. Clinical evolution of HIV infection was faster and more devastating in the homosexual group than in the heterosexual group. PMID- 7482115 TI - [Incidence and characterization of large-cell anaplastic lymphoma]. AB - PURPOSE: Large cell anaplastic lymphoma (LCAL) is a new entity among large cell lymphoma. Diagnosis is based upon morphology and on positivity to Ki1 antigen. The objectives of this study were: 1) to determine the incidence of LCAL in the patient population seen at the Instituto Roffo between 1981 and 1993; 2) to study the immunological phenotype; 3) to study the association with Epstein-Barr virus and HIV; 4) To study the expression of oncogen bcl-2 and 5) to evaluate tumoral growth factor. MATERIAL AND METHODS: The study was done on 1030 biopsies of nodal and extranodal lymphoproliferative diseases, with Hodgkin and non-Hodgkin lymphoma. Of the 1030 consecutive cases, 67 were selected. They revealed pleomorphic cells with sinusoidal disposition. Biotin and avidin peroxidase techniques were used to identify the following antigens: CD30(Ki 1), CD45RO (UCHL1), CD20(L26), CD45(CL), Epstein Barr virus (VEB), human immunodeficiency virus (HIV), CD15, CD68, PCNA, bcl-2 oncogen, vimentin, membrane epithelial antigen (EMA). Of these cases, only 10 revealed strong positive reaction to CD 30 (Ki1), thus they were considered to fulfill criteria to be classified as LCAL. RESULTS: 1. Incidence of LCAL in the lymphoma population under study was 1%. 2. Phenotype was B in 3 cases, T in 3, and macrophage in 1.3. There was 1 case of positive EBV and 3 positive HIV. 4. Eight of 10 LCAL cases were positive for bcl 2 protein. 5. Tumoral growth factor was 57%. Also, it was noted that most cases were secondary to Hodgkin lymphoma, and a few secondary to polymorphic immunoblastic lymphoma, with negative reaction to CD15 and EMA and positive for Vimentin. CONCLUSIONS: 1. Incidence of LCAL amongst patients with lymphoma is very low. 2. Immunological phenotype is varied. 3. There was no significant association with EBV or HIV. 4. Oncogen bcl-2 was demonstrated in most cases. 5. There was a high percentage of cells in proliferation. PMID- 7482116 TI - [Influence of hematopoietic growth factors on transfusion support following autologous bone marrow transplantation]. AB - PURPOSE: To determine the effect of haematopoietic growth factors (HGFs) (G-CSF and GM-CSF) on supporting platelet and red blood cell (RBC) transfusions in patients undergoing autologous bone marrow transplantation. PATIENTS AND METHODS: We retrospectively evaluated the transfusion requirements of 44 patients over three months post-transplant. Ten of these patients received recombinant human G CSF and ten received GM-CSF. This group was compared with the control group, formed of 24 patients who did not received HGFs. The patients receiving HGFs did not differ significantly from those who did not receive growth factor with regard to age, sex, diagnosis, number of bone marrow cells infused and clinical factors affecting the efficacy of platelet and/or RBC transfusion (fever, infection, amphotericin B treatment, bleeding episodes). Statistical analysis of the results was made using the Mann-Whitney U test, with a p value significant at the 0.05 level. RESULTS: No significant effect of HGFs on platelet cell recovery was observed, but there was a trend for the time taken to recovery to increase. The median time of duration of platelet support dependence in the HGFs treated group was 19 days (range 7-100), compared with 15 days (range 3-65) in the control group. The number of platelet transfusions was 8.5 (2-43) and 7.5 (1-33) respectively. The treated patients received a median of 57 units of platelets (range 12-363), compared with 49 in the controls (range 7-206). In NHL there was a reduced need for platelet transfusions in HGFs group. In HD, HGFs increased platelet usage considerably (p < 0.05), although these groups were formed by only 5 and 7 patients respectively. The median RBC usage was the same between both patient groups (6 units of packed red cells). CONCLUSION: In our patients, the administration of HGFs has no beneficial effect on blood product requirements. PMID- 7482117 TI - [Expression of latent membrane protein (LMP) in large-cell anaplastic lymphomas]. AB - PURPOSE: The presence of Epstein-Barr virus (EBV)-encoded latent membrane protein (LMP) was investigated in 40 cases of lymphoproliferative diseases which include Hodgkin's disease (HD), anaplastic large cell lymphoma (ALCL) and non-Hodgkin's lymphoma (NHL) of B and T-cell nature. MATERIAL AND METHODS: All cases were immunophenotyped in paraffin-embedded lymph node tissues, with a complete panel of monoclonal antibodies against B-cells, T-cells, histiocytes, activation and proliferation markers and classified as: 24 anaplastic large cell lymphoma (ALCL, 8 classical type and 16 ALCL-HD related), 10 lymphocyte predominant HD (LP, 5 classical type and 5 with ALCL areas), 4 NHL (two T-Cell type and 2 T-cell rich B cell NHL). Immunohistochemistry techniques were performed ABC-complex and phosphatase alkaline anti-phosphatase alkaline (APAAP). RESULTS: LMP was detected in 35% (14/40) of total cases. In LP group one third of cases were LMP+. In ALCL HD related cases 44% were LMP+ versus 13% in ALCL group. All LMP cases were CD30+ except one NHL-T and a T-cell rich B-cell NHL. The predominant immunophenotype was LMP+/CD20+ (57%) versus LMP+/CD20-. Most cases were of B-cell (36%) lineage. Null ALCL cases were LMP-. CONCLUSIONS: LMP, the most oncogenic EBV protein could play a pathogenic role in lymphoproliferative processes. It is not exclusive of HD and appears in other NHL preferentially of B-cell nature, above all in ALCL cases relating the two neoplasias HD and ALCL, both CD30 positive. PMID- 7482119 TI - [Characterization of mutation of the P53 gene in lymphomas]. AB - PURPOSE: To analyze P53 mutations in a series of NHL and to determine its implication in the inactivation of the tumor suppressor function. MATERIAL AND METHODS: We analyzed 18 lymphomas by SSCP (exons 5 to 9) and subsequent sequencing technics to detect and characterize mutations on this gene. Loss of heterozigosity (LOH) was also studied with a VNTR near the p53 gene and with a intragenic microsatellite, comparing tumor and normal tissue. RESULTS: We found altered bands by SSCP in 5 cases, 4 of them have been described by sequencing analysis. One case was a polymorphism and the others were missense mutations. All mutations appeared in high grade lymphomas. Only one case showed LOH in both VNTR and microsatellite. CONCLUSIONS: This results suggest that p53 mutations may be associated with more malignant lymphomas. PMID- 7482118 TI - [Large-cell anaplastic lymphomas: a genetic and immunophenotypic study]. AB - PURPOSE: To characterize from a genetic point of view a group of non-Hodgkin's anaplastic large-cell lymphomas (ALCL) by Southern blot and PCR methods with different probes (molecular study) and with direct or post 24-78 hours cultures with GTC banding techniques (cytogenetic). To correlate the results to the immunophenotype performed with a complete panel of monoclonal antibodies (MoAb) according to avidin-biotine and alkaline phosphatase (APAAP) methods. MATERIAL AND METHODS: Sixty cases of ALCL were reviewed and only 19 selected (with frozen or fresh material) because a complete immunohistologic and genotypic correlation had been done. According to CD15 expression two groups were considered, CD15+ (the so-called Hodgkin's related or Hodgkin's like) and CD15- or classical type. RESULTS: Only 26.5% of cases showed immuno -genotypic correlation. Immunohistochemistry is an accurate method for activation, proliferation and B cell nature, but T-cell cases were not stained because T-cell paraffin markers are not completely specific. CD15 group had only 30% rearranged cases with scarce cytogenetic abnormalities, as it occurs in Hodgkin's disease (HD). ALCL classical type showed 66% rearranged cases, and one of the T-cell cases showed an incomplete t (2:5) translocation or polyploid cell lines. CONCLUSIONS: Both groups have different genetic and immunophenotypic behaviour which resembles HD or NHL. CD15 positive cases or ALCL HD-related constitute a borderline entity which has to be recognized because of the different therapy and clinical behaviour. PMID- 7482121 TI - [Current clinical use of hematopoietic growth factors]. PMID- 7482120 TI - [Treatment of idiopathic thrombocytopenic purpura with danazol]. AB - PURPOSE: To assess the response to danazol in our patient population with ITP. PATIENTS AND METHODS: We evaluated 17 patients with ITP (11 women and 6 men) with average age 67.3 +/- 9.9 years (range 46-83): 10 patients with acute ITP without response to corticosteroids during 4-6 weeks; 4 patients with acute ITP without treatment; 2 patients with chronic ITP and 1 patient with recurrent ITP resistant to corticosteroids. The dose of danazol ranged between 100 and 600 mg/d. The patients were included in two groups: 1) Responders if they achieved complete or partial remission; 2) Non-responders if they achieved transient or fair response. RESULTS: The overall response rate was 58.8% and in patients previously treated 46.1%. The time to response was 35.6 +/- 48.5 days (range 14-180 days). Among the 10 responder patients 6 have remained in remission, 1 died 6 months after the remission for unrelated disease, and 3 relapsed after discontinuation of treatment with a good response after the reinstitution of therapy. Two patients resistant to treatment with danazol were splenectomized with transient response, and they responded favorably to the reinstitution of danazol. The treatment had a good tolerance. CONCLUSION: 1) The treatment with danazol in ITP is a good alternative therapeutical approach as initial treatment in ITP and ITP patients without response to corticosteroids. 2) ITP patients resistant to danazol may response to this treatment after splenectomy. PMID- 7482122 TI - [Granulocyte abnormal chromatin clumping syndrome: report of a new case and review of the literature]. AB - Abnormal chromatin clumping syndrome (ACCS) is a rare granulocyte disease halfway between Ph'-negative chronic myelogenous leukaemia (CML) and the myelodysplastic syndromes. A new case of such syndrome is presented, the patient being a 65 year old man with anaemia, thrombocytopenia and myelaemic leucocytosis with abnormal condensation of the neutrophils' chromatin. Marked granulocytic hyperplasia was present in the bone marrow with strikingly dysplastic features, along with a decrease of the remaining blood cell lines. Irregular nuclear outlines and heterochromatin clumps separated by euchromatin bands were seen in the more mature granulocytes upon ultrastructural studies. No cytogenetic abnormalities were found in the bone marrow, and no ber-abl gene rearrangement was present in white cells from peripheral blood. The clinical course was marked by persistence of the anaemia and increasing leucocytosis which required 6-mercaptopurine treatment. Acute myeloblastic leukaemia occurred one year later and the patient died shortly after. It was concluded, from this case and others reported in the literature, that, with regard to its clinical course and proliferative character, ACCS is closer to Ph'-negative, bcr-abl negative CML rather than to the myelodysplastic syndromes. PMID- 7482123 TI - [Abundant erythroblastic islands in bone marrow aspirate in a case of polycythemia vera. Various considerations on this anatomo-functional structure]. AB - Despite the initial description, more than 30 years ago, of an erythroblastic island (EI), little is known about the mechanisms of union and interaction between the different cells of this anatomical and functional bone marrow structure. The observation of entire Els is extremely uncommon in normal bone marrow aspirates because of their dispersion during spread. A case report is described in which a 73-year-old woman, diagnosed 20 years ago with polycythaemia vera, showed multiple Els in a bone marrow aspirate. Their immunophenotyping revealed deficiency of several integrins. The ultrastructural study indicated a marked erythroblastic rophalocytosis. A hypothesis is suggested in view of these observations. PMID- 7482124 TI - [Fludarabine and fatal hemolytic anemia]. PMID- 7482125 TI - [Low-molecular weight heparins in dental extractions of patients receiving oral anticoagulant treatment]. PMID- 7482126 TI - [Leukocyte content in platelet concentrates]. PMID- 7482127 TI - [Health assistance hematology at a county hospital]. PMID- 7482128 TI - The distribution of OASDI taxes and benefits by income decile. AB - On average, persons receiving Social Security benefits tend to have lower current incomes than do persons paying Social Security taxes. This article documents OASDI's income distributional patterns by dividing the 1992 Current Population Survey population into 10 income deciles and tabulating benefits received and taxes paid by each decile. The benefits and taxes, when compared with non-Social Security income, are progressive: as income rises from decile to decile, the ratio of benefits to income falls, and, except at the highest deciles, the ratio of taxes to income rises. A large component of the current income distributional pattern is associated with age: the young on average receive more income and pay more taxes; the old on average receive more benefits. However, when benefits and taxes are tabulated for income deciles within specific age groups, a general progressivity is still observable, although it is weaker than that for the population as a whole. PMID- 7482129 TI - A guide to Social Security money's worth issues. AB - This article discusses some of the major issues associated with the question of whether workers receive their money's worth from the Social Security program. An effort is made to keep the discussion as nontechnical as possible, with explanations provided for many of the technical terms and concepts found in the money's worth literature. Major assumptions, key analytical methods, and money's worth measures used in the literature are also discussed. Finally, the key findings of money's worth studies are summarized, with some cautions concerning the limitations and appropriate usage of money's worth analyses. PMID- 7482130 TI - Mortality and income inequality among economically developed countries. AB - The absence of a correlation between age-adjusted death rates and the average income levels of economically developed countries has led researchers to conclude that income does not affect the mortality levels of economically developed countries. The mortality experiences of the former Soviet Union and some of the eastern European countries have further brought into question the importance of income's distribution in determining mortality among economically developed countries; prior to its breakup, the income distribution of the Soviet Union was as equal as that of Sweden, yet the life expectancy of the Soviets has been dramatically shorter than that of the Swedes. Using insights from a longitudinal microanalysis of U.S. mortality, this study presents evidence that, even for economically developed countries, the income distribution of a nation is an important determinant of its mortality. The results of this study also suggest that the relatively unequal income distribution of the United States is an important contributing factor to its low life expectancy relative to other high income countries. PMID- 7482131 TI - Workers' compensation: coverage, benefits, and costs, 1992-93. AB - Benefit payments under workers' compensation programs in the United States during 1992-93 stabilized in comparison with the experience of prior years, which had been marked by substantial growth. In 1992, the total benefit amount of $44.7 billion was 5.9 percent higher than the amount in 1991. The 1993 benefit amount of $42.9 billion represented a 3.9 percent decrease from the amount in 1992. From 1991 to 1993, benefits increased by only 1.8 percent. The payments for 1993 included $25.4 billion in wage-loss compensation and $17.5 billion for medical care. Employer costs of providing workers' compensation was $55 billion in 1992 and $57.3 billion in 1993. The cost of protection per covered employee was $597 in 1993, equal to $2.30 per $100 of payroll. In the same year, there were 96.1 million workers covered under Federal and State programs. This article examines 1992-93 program experience in terms of benefits, costs, and the components of change. PMID- 7482132 TI - Actuarial status of the Social Security and Medicare programs. PMID- 7482133 TI - Occupational risk of infection with hepatitis B and hepatitis C virus. AB - The risk of HBV infection among HCWs is well documented. With widespread use of hepatitis B vaccine, the risk of infection can largely be eliminated. Although some older physicians still have not received the vaccine, increased use by younger physicians and by HCWs in general in response to OSHA regulations is very encouraging. The risk of HCV infection among HCWs is not well defined. Although based on available data, this risk appears to be low, the risk of chronic liver disease with HCV infection is high, and effective measures that prevent transmission and infection are needed. PMID- 7482134 TI - Occupational risk of infection with human immunodeficiency virus. AB - The risk of HIV infection in surgical settings is a composite of overlapping risks related to the local prevalence of HIV, the route of exposure to HIV infected blood, and the susceptibility of the worker. Studies continue to suggest that the risk of blood contact, including percutaneous injuries, remains appreciable. Prevention of such exposures in the operating and delivery room by adoption of safer instruments, work practices, and techniques and by the consistent use of appropriate personnel protective equipment must be viewed as a priority. PMID- 7482135 TI - Microbiologic considerations. Disinfection and sterilization strategies and the potential for airborne transmission of bloodborne pathogens. AB - Bloodborne pathogens are transmitted from person to person by sexual contact and by injection of drugs and, in health-care settings, by needlesticks and, in the case of HBV, by mucous membrane exposure. Conventional disinfection and sterilization and housekeeping strategies are sufficient for the environmental control of bloodborne viruses. Environmentally mediated infection transmission of bloodborne viruses is unlikely and only HBV, because of the extraordinarily high viral concentration in blood, is thought to have a real potential for such transmission. No epidemiologic or laboratory data support the hypothesis that surgeons or other health care workers are at increased risk of infection by bloodborne pathogens. Although theoretically possible, airborne transmission does not appear likely. Studies that claim to show the potential for airborne transmission of bloodborne pathogens ignore the basic principles of aerobiology and the requirements for proving environmental transmission. PMID- 7482136 TI - Are universal precautions realistic? AB - In concluding whether universal precautions are necessary, it certainly appears that we need something to reduce the significant problem of HIV transmission to health-care providers. As occupational risk goes, it exceeds the occupational risk of a number of other high-risk professions. Unfortunately, we do not know if universal precautions are effective. We also do not know the true compliance rate in use of universal precautions, nor whether they have an impact on transmission even if effectively used. What are the alternatives? They are not great, but some have not been adequately explored or implemented. Re-engineering around needle use in the hospital is clearly the most likely area to produce concrete results, because needlesticks are overwhelmingly the greatest source of infection, but this has not been encouraged to the degree it could be, even with systems already developed. Universal testing does not appear to be a viable alternative, for numerous reasons already discussed. Finally, are universal precautions more important for other pathogens than HIV? I would say yes. Hepatitis B, hepatitis C, and nosocomial infections are more important both as public health issues and as health-care provider prevention issues. If universal precautions are effective in reducing any of these, they are worthwhile. PMID- 7482137 TI - Epidemiology of injuries by needles and other sharp instruments. Minimizing sharp injuries in gynecologic and obstetric operations. AB - Surgical and obstetric HCWs and epidemiologists will benefit from working together to describe the frequency and circumstances of percutaneous injuries in operating and delivery rooms. Rates of percutaneous injury vary among institutions, and attending and resident surgeons are among those at greatest risk for injury. Gynecologic surgery appears to have one of the highest rates of injury of the surgical specialties, and rates of injury vary by procedure within a given specialty. Suture needles cause the majority of injuries. Certain actions such as holding tissue while suturing or cutting are associated with a higher risk of injury. Many percutaneous injuries appear to be preventable. Epidemiologic data can be used to develop strategies based on the industrial hygiene model to reduce the incidence of percutaneous injury and to collect and disseminate data on the efficacy of new preventive measures. Potentially safer instruments and suture needles, technique modification strategies, and personal protective equipment such as cut-resistant gloves and finger protective strips are now available. Scientific assessment is needed of these and other new measures to determine whether they will decrease the risk of percutaneous injury, be acceptable to users, be cost effective, and avoid adverse consequences to patients or HCWs. PMID- 7482138 TI - Injury prevention in anesthesiology. AB - Anesthesia personnel routinely perform procedures that put them at risk for contact with blood and body fluids. Some of these exposures are to skin and mucous membranes; many are percutaneous injuries associated with the frequent use of needled devices. The high risk in anesthesia procedures for contact with infected body fluids is associated with a high incidence rate of actual infection with bloodborne pathogens. PMID- 7482139 TI - Hand protection. AB - The traditional purpose of surgical gloves is to prevent transmission of pathogens (usually bacterial) from surgeon to patient. Yet the hand is also the most common site of injury and blood contamination among operating room personnel. Thus, gloves also can protect against transmission of pathogens from patient to surgeon. This article focuses on the value of gloves for hand protection. The current data on such protection derive exclusively from studies that use glove leak and contamination as outcome measures. There are no data that measure protection in terms of actual disease transmission. PMID- 7482140 TI - Prevention of blood exposure. Body and facial protection. AB - Regardless of the specific clinical setting in the operating room, it is clear that better protection of all personnel is an appropriate objective in the current environment. Better protection through improved PPE and modification of operational practices is essential. A prompt response to blood contact when it does occur is likewise appropriate. With conscientious applications of methods to reduce blood exposure, it is hoped that the operating room can become a safer place with respect to occupational infections from bloodborne pathogens. PMID- 7482141 TI - How to select and evaluate new products on the market. AB - New devices and products often promise to protect health-care workers and patients from transmission of viral infections. These need to be evaluated carefully for efficacy, applicability, and cost in an objective, structured manner. PMID- 7482142 TI - Practical considerations in purchasing new products. AB - The introduction of acceptable PPE to OR personnel requires several items: a practical consideration for implementing CDC guidelines, evaluation of fiscal issues, a coordinated effort of clinical trials, and adherence to other standards for OR products. It is essential to know current research in the area of PPE in order to help institutions make knowledgeable, cost-effective, safe decisions on the purchase of products for the prevention of employee exposure to bloodborne pathogens. PMID- 7482143 TI - Postexposure prophylaxis for occupational exposures to hepatitis B, hepatitis C, and human immunodeficiency virus. AB - Bloodborne pathogens are becoming increasingly prevalent in, and therefore contributing increasing levels of risk to, the health-care work-place environment. This problem is magnified in the blood-intense operating room and obstetric environments. Whereas we will never be able to eliminate such risks entirely from the health-care workplace, a multifaceted approach to the management of these risks throughout the hospital environment and particularly in risk-intense environments will likely create a safer milieu and climate. Such an improved environment will clearly be necessary as we continue to strive to provide optimal care for all patients, irrespective of their bloodborne infection status. PMID- 7482144 TI - Preventing bloodborne pathogen transmission from health-care workers to patients. The CDC perspective. AB - The development of recommendations to manage the risk of bloodborne pathogen transmission from health-care workers to patients during invasive procedures has been difficult, primarily because of the limitations of available scientific data. Ultimately, both health-care workers and patients will be protected best by compliance with infection control precautions and by development of new instruments, protective equipment, and techniques that reduce the likelihood of intraoperative blood exposure without adversely affecting patient care. PMID- 7482145 TI - Bloodborne pathogen transmission from healthcare worker to patients. Legal issues and provider perspectives. AB - Health-care providers have an obvious, primary obligation to patients. Yet providers also have obligations to the public health (society), their institutional or individual self-interests, and their employees (fellow health care workers). These obligations contain inherent conflicts, and attempts to reconcile the conflicts often perpetuate contradictions. This article identifies and discusses some of the moral and legal bases of these conflicts. PMID- 7482146 TI - Normal topography of the conus elasticus. Anatomical bases for the spread of laryngeal cancer. AB - The topographic relations of the conus elasticus were studied with special regard to the local spread of laryngeal cancer. Sections of twelve plastinated adult human larynges were investigated. Lateral from the median cricothyroid ligament, the conus elasticus reveals two broad gaps containing adipose tissue and blood vessels. Along these routes, tumors of the larynx may easily extend into the ventral extralaryngeal tissues by continuous growth. Fibres of the conus elasticus cover the entire cranial surface of the vocalis muscle near the muscle's insertion at the thyroid cartilage. This part of the conus elasticus has been termed "thyroglottic ligament" in the fetus [22]. In the adult, this ligament prevents early cancer of the anterior vocal cords from invading adjacent structures. Other collagenous fibres continuous with the conus elasticus provide only an incomplete layer separating the lateral cricoarytenoid and the thyroarytenoid muscle. These fibres do not constitute an effective barrier against cancer growth. PMID- 7482147 TI - Distal vascular pedicle-hemisoleus to tibial length ratio as a main predictive index in preoperative flap planning. AB - The management of severe compound tibial fractures is aimed at obtaining osseous consolidation, proper cutaneous and muscular cover, absence of infection and restoration of limb function. A knowledge of the cutaneous and muscular vascularisation allows the surgeons to use many flaps in a rational and predictable manner. One of the most commonly used flaps is the soleus muscular flap. This may be used as a proximally or distally based muscular flap and the use of a hemisoleus flap has also been described. A morphometric analysis of the relation between tibial length and soleus vascular pattern provides a simple and reliable method for planning this flap preoperatively. The study was performed on fresh and preserved cadavers. PMID- 7482149 TI - Reconstruction of the aortic arches of a 28-day human embryo (stage 13) using the Born technique. AB - We present a reconstruction of the aortic arches of a human embryo at stage 13. Only a few original observations have been reported during the last 110 years and only two with three-dimensional reconstructions. The reconstruction technique originated in 1885. Modern computerized reconstruction techniques have no evident advantages and are more expensive. Aortic arch malformations are responsible for disorders which cannot be understood without a knowledge of their embryology. The embryonic aortic system is a typical example of Haeckel's law of recapitulation. A human embryo at stage 13 was prepared in 5 mm sagittal sections so as to reconstruct the entire dorsal aorta, the second (transient) arch, the third and fourth arches, and the sixth arches responsible for the pulmonary artery, pharynx, larynx and tracheoesophageal axis. They are only 5 pairs of arches in man. At this stage, the most important vessel is represented by the dorsal aorta with a cerebral route, due to predominance of the development of the central nervous system and its vascularisation over other organs. Vascularisation of the lower limbs appears before their macroscopic development and is probably one of the factors responsible for their growth. Many such reconstructions are necessary if conclusions are not to be based on too few specimens to be sound. A better knowledge of embryology lead to a new understanding of aortic arch malformations. PMID- 7482148 TI - A study of the fasciocutaneous vascularisation of the arm. Surgical applications. AB - The data on the fasciocutaneous vascularisation of the medial aspect of the arm are incomplete in the literature. This study presents the methodology and results of an anatomic study on 19 upper limbs with arterial injection. The vascularisation of the upper part of this region is provided in 68% of cases by the superficial fasciocutaneous branch (SFCB) of the superior ulnar collateral artery (SUC), that of the middle and lower part by 3-4 fasciocutaneous pedicles originating from the SUC which are prolonged distally by the recurrent posterior ulnar artery. The SFCB has important relations with the ulnar nerve; it travels dorsal to the nerve in 64% of cases and then allows the raising of a distally pedicled ulnar flap with a 3:1 ratio. In other anatomic configurations, a flap with retrograde flow supplied by the inferior pedicles of the SUC can always be raised. Its ratio is then 2:1. this flap allows coverage of all cases of posterior or anterior losses of substance at the elbow. These results supplement the data required for the raising of a distally-pedicled medial brachial flap, but also of a proximally-pedicled superior medial brachial flap and of a free superior medial brachial flap transposed by microsurgical means. PMID- 7482151 TI - Microsurgical anatomy of the human ophthalmic artery: a mesoscopic study of its origin, course and collateral branches. AB - In the present paper we have studied the gross (mesoscopic) anatomy of the ophthalmic a. in humans, using magnification by microsurgical systems to obtain data on the origin and course of this artery and its main collateral branches. Comparison of our results with previous reports indicates that, although the anatomical variations of the vascular system are well known, some patterns of frequency may be emphasised. Thus, the ophthalmic a. was usually found as a collateral branch of the internal carotid a., although other origins were also found. The ophthalmic a., once inside the orbit, followed a course above the optic nerve in most cases. All the collateral branches of the ophthalmic a., with the exception of the muscular branches, showed great constancy. PMID- 7482152 TI - Preliminary study of the human lumbar spine using speckle technique. Limits of compression, shear and torsion. Speckle photography--electronic speckle pattern interferometry. AB - Optical techniques using single wavelength lasers allow precise study of the superficial displacements of deeper anatomical structures through direct mechanical pull. Two techniques have been used in this study: 1. Double exposure speckle photography. Since 1980 the relationship between the surface displacements and minimal traction applied to the transverse processes of the spine, either unilateral or bilateral, was studied in an attempt to simulate muscular action exerted at this level. 2. Computerised speckle interferometry. This allowed analysis of the deformations between the lumbar vertebrae under vertical compression, shear or torsion outside the usual anatomical planes. This preliminary study outlines the mechanical behaviour and the plasticity of the vertebral structures but has been performed on only a few anatomical specimens thus not allowing statistical analysis. This would require a larger series. PMID- 7482153 TI - Topometry of normal intrahepatic bile ducts. AB - This paper focuses on practical problems which may also prove to be of theoretical importance, by presenting a method of establishing an exact topometry of the intrahepatic bile ducts. Measurements were made on corrosive casts of the intrahepatic bile ducts from 13 human livers. On the basis of the topometric results presented it is possible to construct an adequate model by means of plastic tubes and to use it for the study of bile flow. PMID- 7482150 TI - Blood supply of the olfactory nerve. Meningeal relationships and surgical relevance. AB - The authors report the results of a series of dissections and anatomic sections of the fronto-basal region of the brain and of the anterior cranial fossa in human cadavers. The constant presence of an arachnoidal cistern above the olfactory nerve was verified. The arachnoid separates from the pial membrane and forms a bridge with the ventral part of the olfactory bulb and tract, from the lateral edge of the olfactory sulcus to the medial edge of the gyrus rectus. The cistern is wide in its anterior portion, between the gyrus rectus and the olfactory bulb, and is reduced to a virtual slit in its posterior portion where the tract is lodged in the olfactory sulcus. The olfactory nerve can be separated without damaging fronto-basal arachnoidial adhesions over several centimeters. Dissection of this region after intravascular injection of colored media shows the constant presence of an artery destined to the olfactory bulb and tract. It originates either from the lateral surface of the anterior cerebral a. (segment A2), or from the medial fronto-basal a., and consistently provides terminal branches in front of the olfactory trigone in the medial olfactory sulcus. At their ventral extremity, the olfactory structures are therefore vascularised independently for several centimeters, from the lower face of the frontal lobe. The independent vascularisation of the olfactory nerve, the tenuous and easily detachable adhesions, and the actual presence of a true arachnoidal cistern all contribute to enabling surgical techniques which conserve olfactory function during anterior approaches. PMID- 7482154 TI - The upper end of the femur: dimensions of the endosteal canal. AB - Based on a prospective study of 30 right or left femurs removed in a continuous series in the anatomy department, a study was made of the dimensions of the endosteal canal of the upper end of the femur. The femora were analysed by means of 12 CT scans below the lesser trochanter. The sections were numbered and the dimensional parameters calculated by computer, using an original plan. The results were subjected to descriptive and correlative analysis. From these dimensional findings it appeared that the anatomy of the upper end of the femur is not random; morphotypic models exist and the laws of a normal model are regulated by a mathematical equation. PMID- 7482156 TI - A common bronchial arterial trunk arising from a left subclavian artery: a rare anatomic variant. AB - The origin of the bronchial arteries is very variable. An exceptional case is reported of a common bronchial arterial trunk arising from a left subclavian artery discovered incidentally during angiography in a patient without any pulmonary disease. An embryologic explanation is proposed for this anatomic variant. PMID- 7482157 TI - Excessive anteriorisation of the superior vena cava associated with an azygos lobe. AB - A wide variety of congenital vascular anomalies of the superior mediastinum exist. Being clinically silent, most of these anomalies are detected incidentally on plain radiographs or CT scans where they could be mistaken for mediastinal masses. Familiarity with these anomalies is very important for correct interpretation and avoidance of confusion. We present a case of a mediastinal mass detected accidentally on plain radiography which on further radiological investigation was found to be an unreported normal variant of the superior vena cava (SVC). CT scans of the thorax and superior vena cavograms showed excessive anteriorisation of the SVC in the presence of an azygos lobe. After reviewing the literature and the embryology of the SVC and azygos lobe, we postulate that the variation in the location of the SVC was possibly due to the presence of the azygos lobe. PMID- 7482155 TI - Popliteal artery entrapment syndrome. Anatomic and embryologic bases, diagnostic and therapeutic considerations following a series of 15 cases with a review of the literature. AB - We report 15 examples of popliteal artery entrapment syndrome observed in 11 patients. The anatomical causes were as follows: in one case, the popliteal artery presented an aberrant course medially to the medial head of the gastrocnemius muscle. In 5 cases, there was a small fibrous band linking the medial head of the gastrocnemius muscle to the lateral condyle and crossing behind the popliteal artery; in 5 cases this anomaly was also found in association with an abnormally high and/or internal insertion of the medial head of gastrocnemius muscle. In the last 4 cases, there was a muscular insertion anomaly associated with muscular hypertrophy causing arterial compression. Arteriography performed in the 11 patients showed evocative signs of the diagnosis in all cases where the artery was patent. Two popliteal arteries were occluded. CT scan and MRI examination of the popliteal fossa enabled us to define the muscular origin of the popliteal compression. All of the patients were operated upon; two received a reversed saphenous bypass and all of the others were treated by liberation of the popliteal artery and/or vein by a posterior approach. Follow-up in all patients at long term showed good prognosis. All of the patients were able to take up their previous physical activities without sequelae. Our review of the literature, which is based on 374 cases of popliteal artery entrapment observed in 280 patients, made it possible to define the frequency of the various anomalies observed, their symptoms and the different therapeutic possibilities. The multiple anatomical classifications as well as the arterial and muscular embryology are also described. PMID- 7482158 TI - Unusual association of two unilateral anomalies present in adulthood: pulmonary hypoplasia and renal agenesis. Embryology and clinical expression. AB - We report a case of pulmonary hypoplasia associated with renal agenesia diagnosed in a 46-year-old woman admitted to hospital because of a viral pleuro pericarditis. The chest x-ray raised the suspicion of left pulmonary hypoplasia. The CT scan and the Magnetic Resonance Image confirmed the presence of the congenital lung malformation and detected the absence of the left kidney. The clinical presentation, radiological diagnosis and embryological basis are described. PMID- 7482159 TI - The arterial blood supply of the pancreas: a review. I. The superior pancreaticoduodenal and the anterior superior pancreaticoduodenal arteries. An anatomical and radiological study. AB - The gross anatomy of the pancreatic blood supply has been subjected to numerous studies. The results of such studies, however, have never been summarized in detail, even in the most important textbooks. For this reason, a certain confusion was generated, especially regarding the interpretation of the nomenclature used to identify pancreatic arteries. This review summarizes more than two centuries of studies of the gross anatomy of the pancreatic blood supply, clarifies the arterial nomenclature, and underlines the aspects about which anatomists are not in agreement. Moreover, it supplies, as far as possible, documentary evidence for numerous observations previously reported only verbally. For this purpose, more than 200 references were directly consulted to provide the anatomical background of the topic, and more than 1000 angiograms were studied to support the review with original figures. The present paper, on the superior pancreaticoduodenal and anterior superior pancreaticoduodenal arteries, is the first of a series of articles dealing with the pancreatic blood supply. PMID- 7482160 TI - The Gerald Marks Lecture. SAGES and surgery. Barbers, endoscopists, minimal access surgeons, interventionists. PMID- 7482161 TI - Minimally invasive approach to perforated ulcer--is it? PMID- 7482162 TI - Combined laparoscopic and endoscopic treatment of perforated gastroduodenal ulcer using the ligamentum teres hepatis (LTH) AB - We propose a novel technique for laparoscopic treatment of perforated gastroduodenal ulcers. The principle of this procedure involves the closure of the perforated ulcer using the ligamentum teres hepatis (LTH). The LTH is cut near its umbilical end and then dissected up to the site of its hepatic insertion. The umbilical extremity of LTH is grasped with a Dormia noose passed through the ulcerated perforation via a gastroscope. Using the noose, the LTH is pulled through the ulcerated perforation until its volume fits and completely closes the perforation. This laparoscopic technique was performed in 15 patients (12 M, 3 F) with anterior perforated duodenal ulcer revealed within the previous 6 h. The procedure could not be performed in three cases: diameter of the perforation exceeding 1.5 cm (n = 1), general purulent peritonitis (n = 2). In the other 12 cases, closure of the ulcerated perforation with the LTH was realized without technical difficulty. The postoperative course was uncomplicated. The posttreatment comfort was excellent; the mean period of hospitalization was 10 days (range, 8-14 days). An endoscopic examination carried out following 5 weeks of anti-H2 treatment showed that cicatrization of the ulcer was good and that no pyloric stenosis remained. These initial results suggest that laparoscopic treatment of perforated gastroduodenal ulcer using the LTH is a simple procedure which can be performed with general assurance of success in patients whose perforated ulcers have occurred quite recently. As the laparoscopic procedure is less aggressive than a laparotomy, it enhances the postoperative comfort of patients and prevents the risk of parietal complications.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7482163 TI - Laparoscopic reoperations after failed and complicated antireflux operations. AB - Nineteen patients underwent laparoscopic reoperations for failed or complicated antireflux operations from a total of 248 patients with gastroesophageal reflux disease who had been operated on by this approach. Sixteen had been submitted to open surgery and three to laparoscopic surgery over a period ranging from 5 days to 31 years before the study. Three patients had been submitted to two open antireflux surgeries previously. Seventeen patients had recurrent reflux esophagitis after different types of surgeries, and two patients presented with gastric strangulation after fundoplication. The causes of recurrence were: slipped total fundoplications (3), disruption of total and partial fundoplications (6), too-tight total fundoplication (1), too-low (gastric) partial fundoplication (1), Allison procedure (1), partial fundoplication and paraesophageal hernia (2), and unknown (3). The laparoscopic approach was used in 18 patients and a laparoscopic-thoracoscopic approach in 1. The procedures included laparoscopic total fundoplications (11), partial fundoplications (4), transhiatal esophagectomy (1), Collis-Nissen (1), Roux-en-Y gastrectomy and thoracoscopic vagotomy (1), and intrathoracic fundoplication (1). One patient was converted to open surgery. Intraoperative complications included 1 pneumothorax, 1 gastric perforation, and 1 esophageal perforation during the introduction of a Maloney dilator. Mean operative time was 210 min, ranging from 140 to 320 min. Mean hospital stay was 3.1 days after treatment of failed operations and 22 days after treatment of complications. Postoperative complications included subcutaneous infection (1), gastric fistula (1), and liver hematoma (1). The results have been excellent and good in 84.3% of the patients after a mean follow up of 13 months. We concluded that laparoscopic reoperations are technically feasible with good preliminary results provided that the mandatory expertise is available. PMID- 7482164 TI - Gasless video-assisted reversal of Hartmann's procedure. AB - More than 60% of patients who are submitted to Hartmann's procedure refuse to undergo reversal. This procedure is in fact a major undertaking associated with significantly mortality and morbidity rates. The authors suggest a minimally invasive approach without pneumoperitoneum. A consecutive series of four male patients, average age 64 years, underwent laparoscopic assisted reversal of Hartmann's procedure in our department. The procedure was performed for intestinal malignant occlusion in two cases and for perforated diverticulitis in the other two. Mobilization was nearly immediate and incisional pain almost absent; peristalsis restarted after 36-48 h. Finally, the patients were discharged on day 6. Neither mortality nor morbidity occurred in the 8-month follow-up period. The authors conclude that this new laparoscopic procedure may lead to shorter hospital stays and increased acceptance by patients, while maintaining the same safety of the traditional open procedure. PMID- 7482165 TI - Observations of aggressive, small, flat, and depressed colon cancer. Report of three cases. AB - Three cases of small, flat, and depressed colon cancers are reported. One lesion is less than 1 cm in diameter with lymphatic invasion in the submucosa; the other measures 0.8 cm in the longest dimension and penetrates through the muscular layers to the subserosa. The third one, 1.2 cm in diameter, has had liver metastasis. The endoscopic appearances of two lesions with a resected specimen are presented in color to demonstrate some difficulties in visualizing these lesions for endoscopists. The biologic aggressiveness of these three lesions appears due to their rapid growth, which is expressed by a high mitotic rate of the cancer cells. Their histogenesis is considered to derive from (1) flat adenoma, from (2) serrated adenoma, and (3) from hyperplastic epithelium (or de novo in origin), respectively. PMID- 7482166 TI - Inflammatory markers following laparoscopic and open hernia repair. AB - Laparoscopic surgery may reduce the inflammatory response to surgery by the avoidance of a skin incision which is frequently the site of maximum tissue trauma. We hypothesized that the inflammatory response is less with minimally invasive procedures. The aim of this study was to evaluate the response of inflammatory mediators following laparoscopic and open hernia repair. Thirty-four patients undergoing unilateral primary inguinal hernia repair were prospectively assigned to either laparoscopic mesh hernia repair (n = 14), open mesh hernia repair (n = 11), or a Bassini repair (n = 9). Serum samples withdrawn prior to surgery, 6 h after surgery, and then again at 24 h after surgery were assayed for interleukin-6 and C-reactive protein content. Interleukin-6 levels at 24 h in the laparoscopic (13.1 +/- 3.1 pg/ml), open mesh (15.5 +/- 2.5 pg/ml), or Bassini group) (15.4 +/- 2.0 pg/ml) did not differ significantly. Neither did C-reactive protein levels at 24 h in the laparoscopic (12.4 +/- 2.7 pg/ml), open mesh (23.0 +/- 7.8 pg/ml), or Bassini group 18.6 +/- 6.6 pg/ml) differ significantly. The response of inflammatory mediators to hernia repair is not modified by undertaking the procedure laparoscopically. PMID- 7482167 TI - From laparoscopic training on an animal model to retroperitoneoscopic or coelioscopic adrenal and renal surgery in human. AB - So far, laparoscopic approaches to kidney and adrenal have been limited because of their retroperitoneal location. We here report eight renal and adrenal endoscopic procedures performed in seven patients: two adrenalectomies for hyperaldosteronism, one adrenalectomy for isolated metastasis from an adenocarcinoma of the lung; two nephrectomies for end-stage infected hydronephrosis, two partial nephrectomies for small circumscribed lesions of the kidney, and one endoscopic resection for pain relief of a voluminous cyst at the kidney. The approach was transperitoneal in two cases and retroperitoneal in five cases using the retropneumoperitoneum insufflation technique. One patient was operated by a combined approach using the retro- and transperitoneal routes. All procedures were successfully completed endoscopically. The retroperitoneoscopic approach of the kidney is safe and does not interfere with the peritoneal organs. Its working space is tenuous, but allows a direct access on the kidney with good exposure of its pedicle. For adrenal surgery, the retroperitoneoscopic dissection is more difficult, because movements of instruments are often impaired by the closeness of the costal margin and the iliac crest. However, in case of difficulties we found it very convenient to switch from a retroperitoneal endoscopic approach to a combined coelioscopic and retroperitoneoscopic operation. Far from excluding each other, both approaches are complementary, particularly for difficult situations (i.e., previous peritoneal or retroperitoneal surgery). PMID- 7482168 TI - A financial analysis of laparoscopic and open cholecystectomy. AB - Laparoscopic cholecystectomy (LC) is now the method of choice in treatment of symptomatic gallstone disease. Despite its rapidly growing popularity, comparative costs of this new method and open cholecystectomy (OC) remain unclear. The most outstanding feature of laparoscopic cholecystectomy is the period of short recovery. In Sweden the social insurance office documents sick leave period, sickness allowance, as well as diagnosis and therefore provides a reliable basis for an economic analysis. The purpose of this study was to estimate the hospital cost and costs due to sick leave in a series of patients operated on with elective cholecystectomy using the two methods. In each group 50 consecutive patients were studied retrospectively. The total hospital cost was 10% lower in the laparoscopy group--$1,864 as compared to $2,030 per patient in the OC group. Median number of days off work was 14 after LC and 35 days after open surgery, which corresponds to a median sickness allowance of $516 per patient (LC) compared to $1,424 (OC). Laparoscopic cholecystectomy is more cost effective than open cholecystectomy mainly due to a reduced sick leave period. PMID- 7482169 TI - Herniation of the lung following video-assisted minithoracotomy. AB - Intercostal pulmonary hernia is a rare finding in surgical practice. The hernia presents as an elastic expansion on the outer surface of the chest wall that is usually easily reducible. The protrusion increases in size during expiration and decreases, or disappears, during inspiration. Usually there are no other symptoms. We describe a man who developed a thoracic herniation of the lung 2 1/2 months after a video-assisted minithoracotomy. A concise review of the literature is included. PMID- 7482170 TI - Retained surgical sponge. AB - Surgical foreign bodies are retained more commonly than is suspected. About 50% will become symptomatic in the form of erosion into the bowel or vessels, fistulae, abscesses, obstruction, bleeding, or chronic pain. Expeditious removal is recommended. Laparoscopic retrieval is feasible especially if discovered early. PMID- 7482171 TI - Management of intraabdominal testicle during laparoscopic hernia repair. AB - A 22-year-old man who had previously undergone work-up for right cryptorchidism and been diagnosed as having right testicular absence was referred for repair of a right inguinal hernia. A laparoscopic approach was chosen for this patient, and at the time of laparoscopic herniorrhaphy, a small intraabdominal testicle was visualized and removed laparoscopically. Laparoscopy has served as an excellent procedure for locating intraabdominal testes and planning the most appropriate treatment for many cryptorchid patients. Until recently, atrophic testes located within the abdominal cavity through use of a laparoscope were removed via an open technique. Advances in laparoscopy now enable detection and definitive treatment for these patients without the need for large, more painful incisions. The laparoscopic approach to this patient enabled diagnosis and management of his cryptorchidism and provided a means for repair of his hernia. PMID- 7482172 TI - Possible neoplastic transformation within gastric hyperplastic polyp. Application of endoscopic polypectomy. AB - A total of 263 gastric hyperplastic polyps, which had been removed endoscopically from 202 patients, were clinicopathologically analyzed. Among these polyps, there were nine polyps with neoplastic components (3.4%), corresponding to adenoma in five lesions and mucosal adenocarcinoma in four lesions. Comparing the neoplastic transformed polyps with the pure hyperplastic polyps, there was no significant difference according to age, gender, location, gross appearance, or size. However, the transformed polyps which were located in the lower third of the stomach were larger in size (mean, 20.8 mm) and were more likely to be found among older patients (mean, 75.8 years) than were the pure hyperplastic polyps (mean size and age: 14.5 mm and 61.8 years). These results may indicate the possibility of a different carcinogenesis belonging to gastric hyperplastic polyps by location, and this finding seems to be significant in the application of endoscopic polypectomy. PMID- 7482173 TI - Laparoscopic cholecystectomy following autogenous breast reconstruction with a TRAM flap. PMID- 7482174 TI - Laparoscopic treatment of Meckel's diverticulum. Obstruction and bleeding managed with minimal morbidity. AB - We present two cases in which complication of a Meckel's diverticulum were dealt with, and in one case diagnosed using the laparoscope. One was a large bleeding diverticulum containing ectopic gastric mucosa, with the diagnosis suggested preoperatively, confirmed laparoscopically, and the pathology resected extracorporeally. The second was a partial intermittent small-bowel obstruction due to torsion around the mesodiverticular band, diagnosed and resected via the laparoscope. The literature of Meckel's diverticula and complications is reviewed, with open and laparoscopic treatment options. Although uncommon, many cases of Meckel's diverticulum may be quite suitable for laparoscopic diagnosis and treatment. PMID- 7482175 TI - Treatment of cholecystocolonic fistula by laparoscopy. AB - Cholecystocolonic fistula is an unusual complication of biliary tract disease. Many of the signs and symptoms of these fistulas are nonspecific, so the diagnosis is often not suspected preoperatively. It is important to make the diagnosis then to prevent fecal contamination when the fistula is divided. We recently encountered a patient who, while undergoing laparoscopic cholecystectomy, was found to have a fistula between the gallbladder and the proximal transverse colon. Important features in the management of this case are (1) maintaining a high index of suspicion for the presence of this complication, (2) use of cholecystography to establish the diagnosis, and (3) use of laparoscopic stapling techniques to divide the fistula while preventing fecal soilage. PMID- 7482176 TI - Acute acalculous cholecystitis due to Vibrio cholerae. AB - The case of a 57-year-old woman admitted with symptoms and signs suggesting an intestinal infection caused by Vibrio cholerae, and who also developed a clinical picture compatible with acute cholecystitis, is presented. Cholera was diagnosed by examining a fresh sample of stools and cultures. An abdominal sonogram disclosed signs of acute acalculous cholecystitis. She underwent cholecystectomy, and cultures of a clear fluid and a "milky" sediment found within the gallbladder were also positive for V. cholerae. This microorganism was seen at the gallbladder mucosa microscopically. The strain was serotyped V. cholerae 01 (El Tor) Ogawa and was the etiology of the acute acalculous cholecystitis in this patient. PMID- 7482177 TI - Laparoscopic resection of a large hepatic tumor. AB - Increasing experience among surgeons and a proliferation of endoscopic instrumentation have allowed a greater number of intraabdominal organs to be approached laparoscopically. Procedures once delegated to standard open technique are now attempted laparoscopically with increasing success. The following case report describes the laparoscopic removal of a large segment IV liver tumor. A review of the literature shows this to be one of the most complex hepatic lesions to be removed laparoscopically to date. PMID- 7482178 TI - Prevention of laparoscopic surgeon's thumb. AB - Laparoscopic surgery is being used in an increasing number of operations today. We have been performing various types of laparoscopic techniques in our department. A problem we sometimes have encountered involves a ring of pressure that develops around the surgeon's thumb at the end of the procedure, which is accompanied by an area of paresthesia in the distribution of the lateral digital nerve. We report our own experience and a small review of the literature. PMID- 7482179 TI - Digital nerve compression due to laparoscopic surgery. PMID- 7482180 TI - Monopolar electrosurgery and contact lasers. PMID- 7482181 TI - Laparoscopic treatment of gastroesophageal reflux disease--defining a gold standard. PMID- 7482183 TI - Laparoscopic reconstruction of gastroesophageal anatomy for the treatment of reflux disease. AB - This paper presents the technique and results of an operation that restores normal anatomical and physiological antireflux mechanisms for the treatment of gastroesophageal reflux disease (GERD). The Hill procedure was modified beginning in 1973, evolving into an operation that has been standard in our practice since 1987. Major changes included total fixation of the abdominal esophagus and elimination of phrenoesophageal bundle plication. We began performing the procedure laparoscopically in 1991 and simultaneously began a study to look at our results. This is the first report of the first 44 patients operated on from October 1991 through March 1994. There was one operative complication. Mean follow-up was 14 months. One patient was lost to follow-up and one patient died. There were no long-term side effects. A Visick grading scale was designed to categorize results. Forty graded satisfactory (95%) and two unsatisfactory. PMID- 7482182 TI - The spilled stone. A potential danger after laparoscopic cholecystectomy. AB - The application of laparoscopic techniques in digestive surgery to areas in which there was no previous experience has favored the appearance of new complications and clinical situations that were not observed during the open era. Initial opinion considered that stones left in the abdominal cavity were harmless, and a few clinical and experimental studies supported this opinion. But cumulative reports of cases suggest a potential danger. From 1991 to date, 49 cases of complications related to stones left in the abdominal cavity have been reported with severe complications that required an open surgical procedure. Stone spillage has not always been considered an indication of conversion of laparoscopic cholecystectomy but is now accepted as a source of infrequent but severe complications that may require a reintervention for treatment. Therefore it is recommended that efforts should be made to retrieve all spilled stones; the surgical procedure should be prolonged until this is achieved, in order to reduce one source of unpredictable morbidity. Open retrieval should be considered in selected cases if a large number or large stones are lost. PMID- 7482184 TI - Laparoscopic Nissen fundoplication in children. AB - Laparoscopic surgery for the definitive treatment of gastroesophageal reflux (GOR) has become an accepted alternative to conventional techniques in adults. Although relatively rare, GOR in children represents an important clinical entity with symptoms including failure to thrive, nocturnal dyspnea, and vomiting. This paper details our experience in eight children who have undergone a laparoscopic Nissen fundoplication for failed medical treatment of severe GOR. Particular attention is paid to technical aspects of the procedure and the differences between adult and pediatric techniques are emphasized. The preliminary results suggest that a laparoscopic Nissen fundoplication is a safe procedure which significantly improves reflux symptoms in children and that these results are comparable to those obtained with conventional surgery. PMID- 7482185 TI - Intraabdominal humidity and electromyographic activity of the gastrointestinal tract. Laparoscopy versus laparotomy. AB - The purpose of this study was to observe electromyographic activity of stomach, small bowel, and colon during and early after identical laparoscopic and conventional operations to compare the operative trauma. In nine dogs a cholecystectomy was performed laparoscopically (n = 5) or by laparotomy (n = 4). Analysis of electromyographic activity focused on rhythm, frequency, and amplitudes of slow waves. Furthermore, oxygenation of blood and tissue, intestinal impedance, intraabdominal humidity, and temperature were documented to investigate their influence on slow waves. Open cholecystectomy caused an evident decrease of frequency and amplitude of colonic slow waves in comparison to laparoscopic cholecystectomy. Arrhythmic slow waves were observed only in the stomach during conventional cholecystectomy. Stomach and small intestine showed no significant difference of frequency and amplitude of slow waves in both operation groups. Intraabdominal humidity and intestinal impedance differed significantly in laparoscopy and laparotomy. Laparoscopic cholecystectomy proceeded with a minor abdominal trauma documented by smaller alterations of slow waves. This may be caused by reduced peritoneal desiccation. PMID- 7482186 TI - Hypothermia induced by laparoscopic insufflation. A randomized study in a pig model. AB - Hypothermia is a common postsurgical problem, yet information documenting the impact of laparoscopy on perioperative heat balance is scarce. This paper quantifies the changes in core temperature over a 3-h period of high-flow CO2 insufflation in a randomized, controlled trial of six pigs. Each animal was anesthetized and studied on three occasions under standardized conditions, acting as its own control via insufflation with no gas compared with insufflation by cold gas and warmed gas. Insufflation of CO2 gas at high-flow rates over a prolonged period of time results in a significant fall in core temperature. The provision of warmed rather than cold insufflated gas confers no protection against changes in core temperature during laparoscopic surgery due to the small amount of heat required to warm the gas to body temperature. A much greater effect is the latent heat required to saturate the insufflated gas. Most of the hypothermic effect is due to this, and could be minimized by humidifying the flow. PMID- 7482188 TI - An evaluation of laparoscopic adhesiolysis in patients with chronic abdominal pain. AB - The purpose of this prospective study was to determine whether laparoscopic adhesiolysis ameliorates chronic abdominal pain in patients with abdominal adhesions. Forty-five patients with chronic abdominal pain lasting for more than 6 months but with no abnormal findings other than adhesions found at laparoscopy underwent laparoscopic adhesiolysis. Thirty-six patients (80%) were available for follow-up after a median time interval of 10 months (range: 6-36 months). Seventeen patients (47.2%) were free from abdominal pain and 13 patients (36.1%) reported significant amelioration of their pain. Six (16.6%) patients had no amelioration. Twenty-nine patients (80.6%) judged the outcome of the operation to be good or beneficial and 35 (97.2%) said that they would undergo the operation a second time if that were necessary. Laparoscopy is an effective tool for the evaluation of patients with chronic abdominal pain, and laparoscopic adhesiolysis cures or ameliorates chronic abdominal pain in more than 80% of patients. PMID- 7482187 TI - Laparoscopy without pneumoperitoneum. Effects of abdominal wall retraction versus carbon dioxide insufflation on hemodynamics and gas exchange in pigs. AB - Laparoscopic surgery with CO2 insufflation is associated with adverse effects on hemodynamics and gas exchange. The abdominal wall retractor (AWR) is an alternative for pneumoperitoneum. Hemodynamics and gas exchange during the use of an AWR were compared to those of CO2 pneumoperitoneum. In eight pigs subjected to 1 h of CO2 pneumoperitoneum or abdominal wall retraction, hemodynamics, gas exchange, and oxygen transport were studied in a randomized cross-over study design. The only change observed during abdominal wall retraction was mild respiratory alkalosis. In contrast, during CO2 pneumoperitoneum mean arterial blood pressure increased 13%, central filling pressures doubled, and a small increase in cardiac output was observed. Peak airway pressures increased 50%, end tidal CO2 increased 20%, and respiratory acidosis was induced (arterial pH from 7.46 +/- 0.07 to 7.31 +/- 0.06 and pCO2 from 33 +/- 3 mmHg to 53 +/- 4 mmHg). Arterial PO2 decreased but mixed venous oxygen saturation and oxygen consumption were unaffected. In contrast with CO2 pneumoperitoneum, laparoscopy using abdominal wall retraction was not associated with adverse effects on hemodynamics or gas exchange. PMID- 7482189 TI - Intraperitoneal fluid collection after laparoscopic appendectomy. Sonographic analysis in asymptomatic patients. AB - The aim of this prospective study was to evaluate the frequency of postoperative fluid collection after laparoscopic appendectomy in patients with normal postoperative development. Twenty-eight patients were included. The surgical technique, histological data, and postoperative development during the first postoperative month were recorded. A sonographic analysis was performed on the 5th postoperative day by a radiologist who was not aware of the histological and surgical data. Ten cases of fluid collection were found (37%). The frequency was higher in cases of suppurated appendicitis and significantly higher with associated periappendicitis. Peritoneal irrigation or retrocecal dissection did not influence the occurrence of fluid collection. Postoperative serous fluid collection occurs with a high frequency after laparoscopic appendectomies, and one must be careful in interpreting sonographic analyses in looking for deep abscesses in patients with difficult postoperative development. PMID- 7482190 TI - A porcine model for laparoscopic adrenalectomy. AB - Laparoscopic adrenalectomy has been successfully performed and described in humans. Though it is presently not widely employed it offers tremendous potential for adrenal surgery. It remains, however, an advanced technique requiring demonstration and practice. The aim of this study is to develop a porcine model of adrenalectomy for teaching and advancing the technique applicable to human surgery. Five female pigs weighing 10-20 kg underwent bilateral adrenalectomy by the lateral approach using endotracheal intubation. The left adrenal was resected with the pig in the right decubitus position. The animal was repositioned to left decubitus and the right adrenal was resected. In each case four 10-mm trochars were placed over the respective flanks. There was no need to mobilize the colon or liver (in the pig model). The peritoneum over each gland was incised and the gland dissected in a cephalocaudal manner. The vessels were ligated and divided particularly those from the vena cava. Upon completion blood loss (< 100 cc) and operating times (mean 60 min for the left and 90 min for the right) were assessed and the animal was sacrificed. The glands were weighed (mean 1.23 g R and 1.15 g L) and measured. Using this model, experience is gained in positioning the subject as well as trocars, in fine dissection in a lateral orientation, and in techniques of hemostasis. Although porcine and human adrenal anatomy differ in some details the pig constitutes an excellent model for the development of the technical experience required to perform adrenalectomy in humans. PMID- 7482191 TI - Prospective evaluation of laparoscopic colon resection versus open colon resection for adenocarcinoma. A multicenter study. AB - Laparoscopic colon resection (LCR) has been performed in the United States sine 1990. This procedure has been accepted by many as a reasonable alternative for nonmalignant, colonic, surgical disease, but the laparoscopic approach remains controversial for curative treatment of carcinoma. In this paper, the results of a nonrandomized series of two large experiences of laparoscopic colon resections were performed and followed for 3 1/2 years in a prospective fashion against an equal number of patients who underwent open resection. The setting was several large metropolitan hospitals in San Antonio, Texas. Over 194 patients were involved in this study. Each patient once diagnosed with resectable colonic cancer was allowed to choose their own procedure, laparoscopic or open colon resection, either of which was performed by the authors. Factors considered include age, sex, body habitus, stage of cancer, margins of resection, numbers of lymph nodes retrieved, hospitalization time, and follow-up period. Observations at this time indicate the following: (1) LCR allows for resection comparable to the classical approach, (2) equal numbers of mesenteric lymph nodes can be retrieved, (3) adequacy of margins of resection can be accurately determined by colonoscopy during LCR, and (4) brief follow-up periods show comparable survival and disease-free intervals. It is the conclusion of the authors that with proper training LCR will come to be recognized as a safe, effective surgical option for treatment of selected patients with colon cancer. PMID- 7482192 TI - Laparoscopic extramucosal myectomy with anterior fundoplication (Dor) for esophageal achalasia using intraoperative manometry. AB - Laparoscopic extramucosal myectomy with anterior fundoplication according to the Dor technique was performed on a 24-year-old-woman. Intraoperative inflation of a pneumatic balloon made the operative procedures such as extended submucosal dissection quite easy. Intraoperative gastrofiberscopy was useful for confirming that the remaining mucosal layer was not injured after completion of myectomy. Intraoperative manometry confirmed a complete decompression of the high-pressure zone in the lower esophageal sphincter. Complete relief of the symptoms has been recognized for 6 months after operation without any medication. It is considered that these laparoscopic procedures including intraoperative inflation of a pneumatic balloon, gastrofiberscopy, and intraoperative manometry can be used as a standard operation for esophageal achalasia. PMID- 7482193 TI - Laparoscopy to correctly diagnose and stage metastatic breast cancer mimicking Crohn's disease. AB - We report a case of metastatic lobular breast carcinoma with extrahepatic gastrointestinal disease. On the basis of clinical findings, radiologic investigations, computerized axial tomography, gastrointestinal endoscopy, and gastric biopsy, the diagnosis of gastric and ileal Crohn's disease was made. The correct diagnosis of peritoneal carcinomatosis was made at laparoscopy. This case exemplifies the utility of laparoscopy in establishing the diagnosis and staging for abdominal disease of uncertain etiology. PMID- 7482194 TI - Thoracoscopic excision of bronchogenic cyst of the esophagus. AB - Bronchogenic cyst of the esophagus is an uncommon cause of dysphagia, usually requiring surgical excision by thoracotomy. A case of thoracoscopic excision is described demonstrating reduced morbidity and rapid recovery. PMID- 7482195 TI - Laparoscopic laser lumbar diskectomy. Operative technique and case report. AB - Approximately 300,000 patients each year in the United States undergo laminectomy for disabling lumbar disc herniation. Post-laminectomy hospitalization is 3-7 days and convalescence may be prolonged. As an alternative to laminectomy, we have developed a technique for performing L5-S1 lumbar diskectomy laparoscopically. Using an anterior approach, the intervertebral disc space is opened and the diskectomy is performed under direct videolaparoscopic imaging. After pneumoperitoneum is established, the patient is placed in a steep Trendelenburg position. The small bowel is retracted cephalad and the colon is moved to the left. The iliac vessels are identified visually and by Doppler probe, and the presacral space is dissected in the midline to expose the L5-S1 disc. In the case presented, the disc annulus was opened with the Nd:YAG contact laser, and diskectomy was performed under direct videolaparoscopic vision using standard neurosurgical instruments modified for laparoscopy. The posterior longitudinal ligament can be visualized directly to define the posterior limits of the completed diskectomy. In the case described, pain relief was confirmed immediately after the procedure. The patient was discharged after 2 hospital days, and returned to normal activity in 8 days. PMID- 7482196 TI - Inferior brachial plexus injury during thoracoscopic sympathectomy. AB - A case of inferior brachial plexus injury during thoracoscopic sympathectomy is presented. As this complication occurred during the first operation of the kind in our department in 1991, it is believed that it can be prevented now by supervision of thoracoscopic sympathectomy by experienced surgeons. PMID- 7482197 TI - An intraluminal surgical approach to the management of gastric bezoars. AB - Trichobezoars are difficult to remove endoscopically, often cause nausea and vomiting, and can result in small-bowel obstruction. A patient with a trichobezoar presented to our clinic with symptoms of partial small-bowel obstruction. Multiple attempts at flexible endoscopic removal were unsuccessful. Two large-diameter percutaneous gastrostomies with an inflatable balloon and distal foam-rubber stent to assure intragastric positioning were introduced under general anesthesia. Visualization was provided by a 0 degree panavision laparoscope placed through one of the gastrostomies. The bezoar was removed through the second gastrostomy using standard laparoscopic instruments. The patient made an uneventful recovery. This is the first reported case of percutaneous removal of a trichobezoar. We conclude large-diameter gastrostomies may serve as a port of access for numerous other intraluminal procedures. PMID- 7482198 TI - Insufflation. Complications and recommendations. AB - This article focuses on complications in colonoscopy related to colonic insufflation. Causes of colonic insufflation complications are examined and methods that may be used to minimize complications are proposed. Testing of state of-the-art video colonoscope systems is the basis of this analysis of insufflation complications and the resulting recommendations. PMID- 7482199 TI - New closure technique for lateral operative laparoscopic trocar sites. A report of 80 closures. AB - The reported frequency of incisional hernias, after operative gynecological laparoscopy, at extraumbilical trocar sites is one per 32 puncture sites created by a 12-mm trocar. A new closure technique of suturing with the Grice Needle (Ideas for Medicine, Inc., Clearwater, FL) before removing the trocars was utilized to close 80 lateral trocar sites (42 consecutive laparoscopic myomectomies). The trocar sizes in this study were 12 mm and 18 mm. This is the largest reported series of lateral trocar-site closures. No hematomas or bleeding or incisional hernias have resulted from use of this technique. This closure allows the surgeon to completely close both peritoneum and fascia, of the lateral trocar sites, under direct laparoscopic visualization without the loss of pneumoperitoneum or risk of inadvertent injury to the small bowel. PMID- 7482200 TI - Atraumatic retractor for endoscopic surgery. AB - A novel retractor has been designed to achieve atraumatic retraction of the liver and displacement of hollow viscera during endoscopic surgery. The functional section of the instrument, when locked, forms a silicon-covered uniplanar hook which can be deployed for horizontal lift of solid organs such as the liver or be used to engage bowel for displacement from the operative field. Following laboratory testing, the instrument has been evaluated clinically and its ease of deployment and atraumatic nature have been confirmed. It has been found to be particularly effective for elevation of the thick fatty left lobe of the liver during antireflux surgery. PMID- 7482201 TI - Gallbladder remnant after laparoscopic cholecystectomy. PMID- 7482202 TI - Hospital charges for Nissen fundoplication. PMID- 7482203 TI - Laparoscopic repair of congenital diaphragmatic hernia in a 6-month-old child. AB - This report describes the laparoscopic approach of closure of a congenital left posterolateral diaphragmatic hernia in a 6-month-old boy. The pros and cons of such an approach are discussed. PMID- 7482204 TI - Chronic cholecystitis following hemobilia due to traumatic intrahepatic injury. AB - We report a patient who presented with colicky abdominal pain, hematemesis, and melena following a blunt abdominal injury sustained in an automobile accident. Serologic tests suggested liver dysfunction and computed tomography (CT) revealed dilatation of the intrahepatic bile duct. Duodenoscopy demonstrated hemobilia originating from the duodenal papilla of Vater. Angiography revealed a pseudoaneurysm of the hepatic artery and angiographic embolization sucessfully stopped the hemorrhaging. However, even following the angiographic embolization, the patient remained symptomatic and repeat CT demonstrated thickening of the gallbladder wall and a lesion resembling a blood clot. We strongly suspected cholecystitis and performed a laparoscopic cholecystectomy. Grossly, the resected specimen revealed wall thickening with a congested mucosa and three blood clots; histologically, these changes were consistent with a diagnosis of chronic cholecystitis. The clinical message of this paper is that patients should be observed for this usual complication of hemobilia after liver injury. PMID- 7482205 TI - Laparoscopic liver resection with the water-jet dissector. AB - Laparoscopic liver resection requires careful patient selection. Tumor size and location have a major influence on the feasibility of a laparoscopic operation. Isolation and ligation of blood vessels and bile ducts after selective liver dissection by suitable techniques are important for visual control of the operating field. Since the Jet-Cutter has proven to give excellent clinical results in conventional liver surgery, we carried out laparoscopic liver resections with the Jet-Cutter in six patients. Five tumors were located in the left liver lobe; the fifth was in segment 6. There were no intra- or postoperative complications. The patients were discharged from the hospital after a mean of 5.4 +/- 2.1 days. PMID- 7482206 TI - Video-assisted thoracoscopic suturing of apical bullae. An alternative to staple resection in the management of primary spontaneous pneumothorax. AB - We prospectively studied thoracoscopic suturing of apical bullae in the management of primary spontaneous pneumothorax. From June 1993 to July 1994, we performed 29 such procedures in 27 patients. (All males ranging in age from 15 to 40.) Bullae less than 2 cm in diameter were imbricated (18), while larger bullae were resected and repaired with 3/0 polypropylene suture (11). Postoperative morbidity was minimal. Averaged postoperative parenteral narcotic (Pethidine) requirement was 88 mg, chest drainage was 1.7 (range 1-4) days, and hospital stay was 2.5 (range 1-7) days. There was no recurrence after a mean follow-up of 10 months. In comparison, 32 patients prior to this study period underwent staple resection of apical bulla. Demographic data in the two groups were similar. Averaged Pethidine requirement in the "staple" group was 98 mg, chest drainage was 1.8 (range 1-5) days, and hospital stay was 2.6 (range 1-7) days. There was no statistical difference in the two groups with respect to pain medication, chest drainage, or hospital stay. The technique of thoracoscopic suturing can be easily acquired. In view of the high cost of staple-cutters, endoscopic suturing should be considered as a viable alternative to staple resection of apical bullae in the treatment of primary spontaneous pneumothorax. PMID- 7482207 TI - A posterior gastric approach to laparoscopic splenectomy. PMID- 7482208 TI - Sutured or fibrin-glued laparoscopic choledochojejunostomy. AB - Minimally invasive techniques for choledochojejunostomy offer theoretical advantages for palliating unresectable periampullary neoplasms. Fibrin glue, a biologic adhesive containing concentrated fibrinogen, may obviate suturing and promote healing without stricture formation. We examined the technical ability to perform laparoscopic choledochojejunostomy (LCJ) and the applicability of thrombin-activated fibrin glue in an animal model of biliary obstruction. Domestic pigs underwent laparoscopic cholecystectomy and ligation of the distal bile duct. Three days later, a side-to-side LCJ was performed by intracorporeal sutured anastomosis (n = 7) or using four stay sutures and homologous fibrin glue (n = 7). Control animals underwent a similar bypass via open laparotomy (n = 7). The postoperative interval to ambulation and oral intake was recorded, and serial serum liver enzymes were measured. The animals were sacrificed at 6 weeks, and tensile strength of the anastomoses was assessed by tensometry. Liver function tests returned to normal values within 7 days following all methods of choledochojejunostomy. In the fibrin glue group, three anastomotic leaks (43%) occurred in the 1st postoperative week. At 6 weeks, all other anastomoses were intact and patent by cholangiogram, but there was moderate stenosis of two open and one fibrin-glue anastomosis. The sutured LCJ, while taking approximately 1 h longer to perform (P < 0.05), resulted in similar efficacy and more rapid recovery (P < 0.05) than open biliary-enteric bypass. Fibrin-glued LCJ was performed rapidly, but had less tensile strength (P < 0.05) and often leaked in the early post-operative interval. We conclude that while there may be a role for laser-activated solders for primary anastomosis, thrombin-activated fibrinogen cannot be advocated as the primary means of creating biliary anastomoses. Using intracorporeal suturing techniques, laparoscopic choledochojejunostomy may be performed safely. PMID- 7482209 TI - The laparoscopic cholangiogram of doom. PMID- 7482210 TI - Incisional dilatation of a stenosed stapled coloproctostomy. PMID- 7482211 TI - Converting from carbon dioxide to nitrous oxide pneumoperitoneum. PMID- 7482213 TI - Changing strategies in the performance of laparoscopic Nissen fundoplication as a result of experience with 230 operations. AB - From September 1991 to January 1995 we performed 230 Nissen fundoplications by a laparoscopic technique. Whilst a loose 360 degree fundoplication secured by 3 or 4 sutures was performed in all instances, there was variation between surgeons regarding the performance of hiatal repair and division of short gastric vessels; 207 operations were completed laparoscopically and 23 were converted to an open operation when a satisfactory wrap could not be achieved. Operating time ranged from 30 to 260 min (median 95) and the median postoperative stay was 3 days (1 19). Twenty-three patients (10%) underwent a subsequent operation (14 within 3 months of the original surgery), 2 for recurrent reflux, 10 for para-esophageal herniation, 2 for a misplaced fundoplication resulting in gastric obstruction, 7 for persistent dysphagia (4 due to stenosis of the esophageal hiatus), 1 for bleeding, and 1 for mesenteric thrombosis. (This patient died.) Five other patients were readmitted to hospital subsequent to their discharge--four because of pulmonary emboli and one because of gastric obstruction. Some 226 patients (98%) are free of reflux symptoms with follow-up ranging up to 40 months (median 16). Absence of reflux and the integrity of the fundoplication has been confirmed by postoperative esophageal manometry and pH monitoring in 90 patients, and by barium meal in 126. Postoperative recovery has been quick and wound-related morbidity minimal. Although the rate of surgical revision was significant in this series, the likelihood of complications or further surgery, as well as incidence of conversion to open surgery, decreased in the second half of the experience.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7482212 TI - Laparoscopic procedure for suspected appendicitis. A prospective study in 283 consecutive patients. AB - Between September 1990 and December 1993, 283 consecutive patients were admitted with clinical symptoms of acute appendicitis. These patients underwent primary laparoscopic approach so that an appendicectomy could be performed by this method. In 49 cases (17.3%), primary laparoscopic examination corrected the preoperative diagnosis and the appendix was left in situ. Appendicectomy was performed in 234 cases (149 women, 85 men) with a mean age of 30 years. Requirement for open surgery occurred in 29 cases. The main cause of unsuccessful procedures was inflammation due to local or generalized peritonitis. Median operative time for a successful procedure was 60 min (range, 25-160). Four postoperative complications (one related to laparoscopic procedure), one case of wound infection, and no mortality resulted. After laparoscopic appendicectomy, the median hospital stay was 3 days (range, 1-16). These results suggest that a laparoscopic approach for suspected appendicitis is reliable, allowing abdominal exploration and safe appendicectomy. PMID- 7482214 TI - Individual assessment of visual perception by surgeons observing the same laparoscopic organs with various imaging systems. AB - Laparoscopic surgery necessitates that operations be performed via a television screen; therefore, image quality is crucial in determining the final outcome. Electronic imaging systems are constantly undergoing revisions and incorporating new ideas. Recently, we have been confronted with a variety of systems, and we, as operators, have no idea what the differences are between the old and the newer systems. As clinicians, we have no electronic yardstick available to compare the options presented or to check the specifications. This study examines critical aspects of image perception. General surgeons, gynecologists, operating-room nurses, as well as allied health personnel all experienced in laparoscopic surgery participated as test subjects. A blinded study was performed in which various camera systems (single chip, three-chip, digitized and nondigitized) were displayed in random fashion. Porcine abdominal areas were displayed using the various imaging units. Participants graded image perception based on the following characteristics: sharpness, color, contrast, and depth of field. The audience were blinded to the types, brand, and number of cameras utilized. A significant preference was shown for digitized systems (P < 0.0001). Also, digitized single-chip cameras scored higher than three-chip cameras (P < 0.05). We propose thorough testing by surgeons and nursing personnel before deciding what type of equipment to purchase. PMID- 7482215 TI - The complications of cholelithiasis caused by state authorization delays. AB - The impact of delay in elective cholecystectomy caused by preoperative state authorization requirements was retrospectively studied. The Student's t-test was used for statistical analysis. From 10/92 to 5/94, 146 patients were evaluated in an outpatient surgery clinic and elective cholecystectomy was planned. These patients were divided into two groups. Group 1 (101) patients waited for state authorization prior to cholecystectomy while group 2 (45) patients did not need state authorization. Group 1 patients waited an average of 112 days from the time of diagnosis to the time of surgery while group 2 patients waited an average of 21 days (P < 0.001). Group 1 patients were more likely to develop acute complications of cholelithiasis and require admission and urgent cholecystectomy than group 2 patients (22% vs 11%, P < 0.05). Group 1 patients requiring emergent treatment were found to have a longer hospital stay (P < 0.001), longer operative time (P < 0.05), and higher intraoperative blood loss (P < 0.005) than group 1 elective cases. Elective cholecystectomy patients who require state authorization have to wait longer prior to cholecystectomy and frequently develop acute complications of cholelithiasis requiring hospital admission and nonelective surgery. This translates into increased patient morbidity and added cost. State authorization requirements act as a barrier to providing timely care, add to cost, and must be considered in the current deliberations on health care policy. PMID- 7482216 TI - The influence of intraoperative gallbladder perforation on long-term outcome after laparoscopic cholecystectomy. AB - During laparoscopic cholecystectomy, gallbladder perforation with leakage of bile and/or gallstones into the abdominal cavity occurs frequently. When this occurs, our practice has been to lavage the operative field and retrieve as many gallstones as possible. We were concerned, however, that complications secondary to infection or adhesions might develop. To address this issue, our first 250 consecutive patients undergoing laparoscopic cholecystectomy were surveyed by postal questionnaire. In the 35-48 months (mean, 41 months) since operation, six patients (2.6%) died of nonbiliary causes. Of the 225 patients (90%) who completed the questionnaire, 73 (33%) suffered intraoperative gallbladder perforation. There were no late wound or intraabdominal infectious complications and no patient has required reoperation for intraabdominal sepsis or bowel obstruction. In the entire group, gastrointestinal symptoms were prevalent and included flatulence (40%), loose stools or fecal urgency (35%), belching (23%), and nausea (4%). The prevalence of these complaints was similar in patients with and without gallbladder perforation. Intraoperative gallbladder perforation during laparoscopic cholecystectomy, therefore, does not cause adverse long-term complications when accompanied by operative lavage and stone removal. PMID- 7482217 TI - A prospective cost and outcome comparison of inguinal hernia repairs. Laparoscopic transabdominal preperitoneal versus open tension-free preperitoneal. AB - We previously showed that patients undergoing transabdominal preperitoneal laparoscopic inguinal herniorrhaphy (TAPP) returned to activity twice as fast as open herniorrhaphy without mesh but that TAPP was twice as expensive. However, it was not clear if the immediate postoperative benefits offered by TAPP resulted from smaller incisions and less tissue dissection or from the requisite tension free placement of mesh. We have therefore completed a prospective outcome and cost analysis comparing TAPP (n = 59) to open preperitoneal mesh herniorrhaphy (PPO) (n = 40) to determine the differences between the two different surgical techniques. When comparing unilateral repairs, there was no difference in hernia type. PPO patients were older (P < 0.05) and their operations were shorter (P < 0.01). Comparison of outcome parameters of pill days, out-of-house activity, and intial day to full activity revealed no difference. Cost analysis showed that total costs, disposable equipment costs, and operating room time costs were significantly less for PPO (P < 0.01). There were two major complications (3%) and twelve minor complications (20%) in the TAPP group while PPO exhibited no major and five minor complications (12%). Follow-up data revealed one recurrence in the TAPP group. There were no recurrences in the PPO group at only 7 months average follow-up. We conclude that since both procedures had similar outcomes in the immediate postoperative period, the increased cost of TAPP and increased potential for both major and minor complications make it difficult to justify its routine use. PMID- 7482218 TI - Laparoscopic hernioplasty. TAPP vs TEP. AB - This study compares the results of two laparoscopic hernioplasties: the transabdominal preperitoneal (TAPP) and the totally extraperitoneal (TEP). Over a 43-month period 1,115 laparoscopic hernioplasties, 733 TAPP and 382 TEP, were performed in 866 patients. There were 11 major complications in the TAPP group (2 recurrences, 6 trocar hernias, 1 small-bowel obstruction, 1 trocar, and 1 dissection injury of the small bowel) compared to 1 recurrence and no intraperitoneal complications in the TEP group. Five TEP procedures required conversion to the TAPP approach, resulting in one umbilical hernia. The median time to return to work did not vary with the approach, but was prolonged in patients compensated for time off, 16 vs 8 days for noncompensated patients. Results suggested that both techniques shortened recovery and eliminated most early failures, but the totally extraperitoneal approach reduced the potential for intraperitoneal complications and may be the procedure of choice in most situations. PMID- 7482219 TI - Pretherapeutic assessment of resectability in patients with upper gastrointestinal tract cancer by using a combination of endoscopic ultrasonography (EUS) and laparoscopy. AB - An accurate pretherapeutic assessment of resectability in patients with upper gastrointestinal malignancies (UGIM) is mandatory in order to choose the optimal treatment strategy. Endoscopic ultrasonography (EUS) has significantly reduced the need for exploratory laparotomy in patients with UGIM, but the pretherapeutic evaluation in about 10% of the patients is incomplete due to certain limitations of the EUS. We prospectively evaluated the use and results of diagnostic laparoscopy in patients with UGIM selected for this procedure by EUS. In six patients with incomplete EUS, laparoscopy demonstrated nonresectability in five patients and a resectable tumor in one patient, and laparoscopy thus filled the informational gap in all cases. In addition, laparoscopy confirmed nonresectability in ten patients in whom EUS had suggested nonresectability. By employing the combinated use of EUS and laparoscopy it seems possible to avoid a great number of futile laparoscopies, and it should also reduce the need for explorative laparotomies. Larger prospective studies have been initiated and might be able to confirm this. PMID- 7482220 TI - Laparoscopic-assisted resection of a gastric lipoma. AB - Laparoscopic excision of a gastric lipoma is reported here. The diagnosis was done preoperatively be computed tomography, which gave evidence of fat tissue due to its low attenuation. The patient undergoes a complete submucosal excision enjoying the benefit of the laparoscopic approach with rapid postoperative recovery. This procedure offers a good alternative to open laparotomy in the treatment of benign gastric tumours. PMID- 7482221 TI - A double or bilobar gallbladder as a cause of severe complications after (laparoscopic) cholecystectomy. AB - A double or bilobar gallbladder is a rare congenital anomaly. If not recognized during preoperative evaluation or operation, it can cause severe complications. We describe two cases in which a second operation had to be performed because of the presence of a second or bilobar gallbladder that was not recognized in the preoperative evaluation and during (laparoscopic) cholecystectomy. The types of anomalies, the concomitant pathology, and treatment are discussed. PMID- 7482222 TI - Stress proteins: their role in the normal central nervous system and in disease states, especially multiple sclerosis. AB - Stress proteins are constitutively expressed in normal CNS tissues, in a variety of cell types (oligodendrocytes, astrocytes and neurons). Their function is uncertain but they may be critical during nervous system development and may protect cells from various stresses, such as hypoxia, anoxia, and excessive excitatory stimulation. Increased amounts of stress proteins are expressed in various cells of the CNS during acute toxic-metabolic states and in more chronic degenerative diseases. Increased expression of stress proteins may constitute a sensitive marker of cell injury. Antibodies to mycobacterial stress proteins bind to normal human myelin and to oligodendrocytes in regions of MS demyelination. Cellular immune responses to stress proteins occur with increased frequency and magnitude in persons with MS, especially those with recent onset of disease. In addition, there are populations of T cells expressing gamma/delta T cells in the brains and CSF of persons with MS, suggesting an in situ immune response to hsp. Humoral immune responses to stress proteins are found in CSF, but no disease specificity has been documented. Some myelin proteins have sequence homology with particular stress proteins. One instance is the homology between a peptide of mycobacterial hsp65 and the myelin protein CNP. Preliminary observations suggest that immune responses to such cross-reactive epitopes modify the course of EAE. All in all, these data support the hypothesis that an immune response to the stress proteins of an infectious agent could result in a cross-reactive immune response to CNS myelin, resulting in demyelination. This may be an especially important mechanism in MS. PMID- 7482223 TI - Immunological findings in amyotrophic lateral sclerosis. PMID- 7482224 TI - Guillain-Barre syndrome: clinical and immunological aspects. AB - Immune-mediated PNS disorders comprise a significant segment of diseases of the nervous system. Studies on GBS as a prototype of these disorders and its experimental model EAN have helped to elucidate some of the mechanisms responsible for myelin injury in the PNS. These mechanisms, although partially understood have been useful in implementing therapies such as plasmapheresis and IVIG and various other immunomodulators. The question of whether an infectious agent such as a virus can directly damage the myelin sheath and/or Schwann cells or whether the agent triggers an immune response against self through antigenic mimicry remains unanswered. The association between C. jejuni infection and GBS has opened new areas of investigation in understanding the immunopathogenesis of the disease. Similar observations with other environmental factors may be made in the future, pointing to the possibility that GBS may not be caused by a single agent but could be the result of an immunological attack on the PNS myelin assembly by a variety of agents or factors. Regardless of the etiology, if the myelin injury is aggravated by product of the immune cells, such as various cytokines, neutralization of these factors could help lessen the burden of injury to the nerves. Future research in autoimmune disorders of the PNS needs to focus on identifying environmental factors that directly, or indirectly through antigen mimicry, damage the PNS myelin. In parallel, further understanding of the immunopathogenesis by dissecting the immunological phenomenon at the systemic and local levels, especially the role of cytokines, growth factors, and adhesion molecules will pave the way for more rational therapies, even if the causative factors are not known. Studies in laboratory animals have demonstrated the efficacy of selective immunotherapy through modulation of the trimolecular complex, i.e., T cell receptor, MHC/molecule, and antigen. Immunological tolerance, presumably through deletion of autoreactive clones, clonal anergy, or active suppression, has proven effective in animals. Other modes of immunotherapy such as nonspecific depletion of T or B cells or down-regulation of activated cells have also been shown to abolish or decrease the severity of experimental autoimmune neurological disorders, including EAN. These immunotherapeutic modalities may become applicable to human autoimmune neuropathies. PMID- 7482225 TI - Immunopathology of the Lambert-Eaton myasthenic syndrome. AB - LEMS is an antibody-mediated autoimmune disease that can occur in isolation, or as a paraneoplastic disorder in association with SCLC (60% of patients). The underlying defect is a reduction in the quantal release of the neurotransmitter ACh from the presynaptic nerve terminal at the neuromuscular junction. Experimental evidence indicates the autoantibodies are directed against nerve terminal VGCCs causing down-regulation in the number of functional channels by cross-linkage. Functional VGCCs have been detected in SCLC cell lines. In cancer associated LEMS it appears likely that antibodies initially provoked by tumour VGCCs cross-react with VGCCs at the nerve terminal, causing the clinical disorder. Antibodies against L-, N- and P-/Q- subtypes of the calcium channels have been identified and radioimmunoassays have been developed to help diagnose the disease. Using peptide toxin 125I-omega-CmTx MVIIC to label P-/Q-type VGCC solubilised from human cerebellum, positive antibody titres can be detected in 85% of patients. However, autoantibodies in LEMS are heterogenous; the antigenic targets include different VGCC subtypes, the intracellular beta subunit and the synaptic vesicle protein synaptotagmin. The disease phenotype may reflect the diversity and titre of these different antibodies. PMID- 7482226 TI - Systemic lupus erythematosus: immunopathogenesis of neurologic dysfunction. AB - Neurologic complications of systemic lupus erythematosus (neuro-SLE) are common. The most frequent manifestations of neuro-SLE are seizures, encephalopathy, and behavioral changes, but a wide variety of other neurologic abnormalities affecting the central and peripheral nervous system and muscle also occur. Although the prevalence of neuro-SLE is high, the diversity of clinical presentations, the multiple potential etiologies, and the absence of sensitive and specific diagnostic tests render diagnosis difficult. Recent advances in understanding mechanisms of neuronal dysfunction combined with advances in imaging techniques, including functional imaging, should help in diagnosis and management. The mechanisms of neurologic injury can be divided into three broad categories. First, neuronal dysfunction may result from direct effects of the immune system on brain cells such as autoantibody binding to cell surface, immune complex deposition with secondary inflammation, and effects of cytokines. Second, immune- mediated injury to supportive structures such as the vasculature may also affect the nervous system by producing ischemia. Finally, the neuraxis may be affected by any one of several immune and non- immune effects of infection, toxins, and metabolic disturbances. PMID- 7482227 TI - Antigen-specific therapies for the treatment of autoimmune diseases. PMID- 7482228 TI - Are current immunological concepts of multiple sclerosis reflected by the immunopathology of its lesions? AB - Immunopathological studies on multiple sclerosis (MS) brain clearly indicate that a T cell-mediated immune response is the driving force in the induction of the lesions. This T cell-mediated response alone, however, is not sufficient to explain the widespread and selective destruction of myelin sheaths. According to present evidence, it is likely that antibodies directed against surface components of myelin sheaths are at least one factor involved in the demyelinating process. The patterns of inflammation, demyelination and oligodendrocyte destruction, however, suggest that the pathogenesis of the lesions may be fundamentally different in individual MS patients and that autoimmunity may not be the sole cause. In the case of autoimmune reactions various different proteins of the nervous system may become targets and it appears unlikely, that myelin basic protein is a major candidate for a pathogenetic role in MS. PMID- 7482230 TI - Percutaneous endoscopic lumbar discectomy: minimally invasive, but perhaps only minimally useful? AB - BACKGROUND A few recent studies dampen the euphoric reports of the value of percutaneous endoscopic lumbar discectomy (PELD) in the treatment of discogenic disease. We felt that a large-scale comparison between PELD and the still minimally invasive open lumbar disk surgery (OLDS) was called for. METHODS This report is based on the surgical experience of one year (1991) and is confined to the intervertebral disk level L4-5. Three hundred thirteen patients treated by open disk surgery were compared with 13 who were selected for percutaneous discectomy. RESULTS Using careful selection criteria, only 13 (4%) of a possible 326 were considered potentially suitable for PELD. Of these, only eight were wholly suitable and were operated on percutaneously. Within the first postoperative month, 62.5% (5 patients) of the PELD group required open surgery for definitive treatment, whereas only 14 (4%) of the 313 OLDS patients had to undergo additional surgery. CONCLUSIONS Although it may of benefit to a very few, we feel that the PELD method cannot be considered a substitute for, or even an alternative to, traditional surgery in most cases. PMID- 7482229 TI - Multiple sclerosis: a unique immunopathological syndrome of the central nervous system. PMID- 7482231 TI - Video-assisted thoracoscopic resection of intercostal neurofibroma. AB - BACKGROUND Video-assisted thoracoscopy (VAT) is being increasingly utilized by thoracic surgeons as an alternative to thoracotomy for several thoracic disorders. Neuroendoscopy is an exciting addition to the neurosurgical armamentarium. These procedures are attractive alternatives in the era of minimally invasive surgery and cost containment, while providing the highest quality medical care to patients. METHODS We report the application of this technology in a patient presenting with intractable thoracic radicular pain secondary to an intercostal neurofibroma. Complete excision of the intercostal neurofibroma was performed utilizing VAT. RESULTS The patient reported good relief of her preoperative thoracic radicular pain. She was ready for discharge from the hospital within 72 hours of surgery. CONCLUSIONS With the advent of improved instrumentation, video-assisted thoracoscopy offers a safe alternative to thoracotomy and the potential benefits of less postoperative discomfort and shorter hospital stays. The potential neurosurgical applications of VAT should not be overlooked. PMID- 7482232 TI - Clinicopathologic growth factors of acoustic neuromas. AB - BACKGROUND Understanding the tumor growth rate is very important when considering strategies for the treatment of an acoustic neuroma, although the natural course of acoustic neuromas has not been reviewed in detail. METHODS The clinicopathologic features and the postoperative growth of tumors were evaluated in 32 patients with acoustic neuromas. This study was undertaken to assess the variability of the growth potential of cells within an acoustic tumor and to determine the relationships between the growth rate and the clinicopathologic characteristics of the patients with acoustic neuromas, including age at surgery, gender, tumor location and preoperative size, the duration of the symptoms, the presence of cystic regions, the presence of Antoni type A and B cells, the tumor cell density, tumor vascularity, mitotic rate, the presence of hyaline degeneration and hemosiderin deposition, nucleolar organizer regions (NORs), and the level of proliferating cell nuclear antigen (PCNA). RESULTS The growth patterns of the tumors were divided into three groups according to their growth rate: a regression group, a "no-growth" group (growth rates from 0-0.11 cm/year) and a progression group (growth rates from 0.19-1.72 cm/year). An additional operation was required in all patients whose growth rate was more than 0.38 cm/year. A statistical study on the factors associated with an increased growth rate showed that the three histopathologic factors most significantly associated with a postoperative growth rate were hyaline degeneration (p < 0.05), cell density (p < 0.005), and PCNA labeling index (p < 0.005). CONCLUSIONS These results strongly suggest that acoustic tumors can be subdivided into several groups, based upon different biologic activities and tumor growth rates. PMID- 7482233 TI - Cerebrospinal fluid leak after acoustic neuroma surgery. AB - BACKGROUND: CSF rhinorrhea is a common complication of acoustic neuroma surgery. METHODS: Twenty cases of persistent cerebrospinal fluid rhinorrhea, 2 of transient leaks, and 2 of late meningitis occurring in a series of 156 posterior fossa acoustic neuroma removals were studied. Prophylaxis of leaks was usually attempted by filling opened air cells in the drilled internal auditory canal (IAC) with bone wax. Communications were repaired by rewaxing the IAC, or by fat packing of either the mastoid or the eustachian tube, middle ear, and mastoid. RESULTS: Most leaks were diagnosed within 1 week of surgery, although 1 occurred 4 years postoperatively. Those seen later were usually recurrences of a previously repaired leak or cases of meningitis. Leaks were more common later in the series when a more lateral unroofing of the IAC was used. The communication usually occurred through air cells opened while unroofing the canal, but two were via the vestibule and oval window. Treatment failed in 38% of cases in which canal rewaxing was used. There were no failures when the communication was sealed by fat obliteration of the eustachian tube, middle ear, and mastoid. CONCLUSIONS: Leaks are more common with a more lateral unroofing of the IAC. A leak may occur into the middle ear via the oval window. Optimal treatment is packing of the middle ear and mastoid with fat. PMID- 7482234 TI - Results and expectations with image-integrated brainstem stereotactic biopsy. AB - BACKGROUND: The histologic diagnosis of an intrinsic brainstem tumor remains problematic due to controversies in methods of biopsy. Despite the widespread use of biopsy techniques, stereotactic brain stem biopsy has received limited attention due to potential morbidity and limited sample size. METHODS: To evaluate the safety and efficacy of brain stem stereotactic biopsy using a dedicated computed tomography (CT)-stereotactic operating room suite, we reviewed our outcomes in 40 consecutive patients over a 13-year interval. This study included patients with midbrain lesions (n = 20), pontine lesions (n = 18), and medullary lesions (n = 2). Midline lesions were approached via a coronal, transthalamic trajectory; lateral brain stem lesions usually were approached via a transcerebellar route. RESULTS: A histologic diagnosis was achieved in 38 patients (95%). All patients had an immediate, intraoperative, postbiopsy CT scan to check for hemorrhage (none occurred). Morbidity was limited to one patient (2.5%) who developed a transient diplopia; there was no mortality. CONCLUSIONS: Stereotactic biopsy for intrinsic brain stem lesions proved as safe and effective as biopsy in the supratentorial compartment. Using high-resolution stereotactic imaging, an appropriate intraparenchymal trajectory, limited sampling, and specific neuropathologic tests, stereotactic techniques within the brain stem were performed with low risk and high accuracy. Biopsy results facilitated specific management strategies for each patient. PMID- 7482235 TI - Stereotactic biopsy of brainstem mass lesions. AB - BACKGROUND: Appropriate therapy of brainstem lesions is guided by accurate diagnosis. Because the majority of brainstem lesions are not amenable to surgical resection, stereotactic biopsy is an attractive method of obtaining pathological tissue. METHODS: We reviewed the medical records of all patients who underwent stereotactic biopsy of brainstem mass lesions at our institution by the senior author (WAF) over a 10-year period ending in December 1993. RESULTS: Twenty-four patients ranging in age from 3 to 68 years underwent stereotactic biopsy of brainstem mass lesions at our institution by the senior author (WAF) over a 10 year period ending in December 1993. RESULTS: Twenty-four patients ranging in age from 3 to 68 years underwent stereotactic biopsy of mass lesions of the brainstem. Sixteen lesions were located primarily in the pons, 7 in the midbrain, and 1 in the medulla. Twenty-two of the biopsies were approached transfrontally and two were approached via the suboccipital transcerebellar route. Pathological diagnosis was made in 23 of the 24 patients. The histologic diagnosis was astrocytoma in 16 patients, metastasis in 3, lymphoma in 1, germinoma in 1, chordoma in 1, progressive multifocal leukencephalopathy in 1, and was nondiagnostic in 1. Complications included 1 case of increased hemiparesis, 1 case of obstructive hydrocephalus, and 1 death. Six patients were less than 20 years of age and in each of these patients the preoperative diagnosis was astrocytoma. In all of these patients the pathology revealed astrocytoma. In the adult patients the pathology was more varied, with 7 of the 18 patients having tumors or pathology other than astrocytoma. CONCLUSIONS: The data suggest that, in adult patients, brainstem lesions are of varied pathology and stereotactic biopsy can provide adequate tissue for diagnosis. The data also suggest that the diagnosis of brainstem glioma in children can often be made without submitting the patient to the risk of surgery. PMID- 7482236 TI - Endoscopic removal of organized chronic subdural hematoma. AB - An endoscopic surgical approach to organized chronic subdural hematoma is described. Advantages of the endoscopic approach include access to virtually the entire hematoma cavity through a small craniectomy performed with local-standby anesthesia. Two illustrative cases are presented. PMID- 7482237 TI - Carbon dioxide gas replacement of chronic subdural hematoma using single burr hole irrigation. AB - In order to achieve complete irrigation, reduce air in the hematoma cavity, and obtain rapid reduction of the cavity without drainage, we use burr-hole evacuation of the hematoma followed by irrigation using physiologic saline solution and replacement of the hematoma with carbon dioxide (CO2) gas. We have treated 19 cases between the ages of 41 and 84. With the exception of one case with dementia of 5 years' duration, all cases were asymptomatic postoperatively. Computed tomography (CT) scans obtained immediately postoperatively showed the gas to fill the hematoma cavity, but on the following day, the gas and the hematoma cavity had nearly disappeared. There was no surgical mortality nor morbidity and there were no side effects of the CO2 gas. Recurrence was found in one case (5.3%). By filling the hematoma cavity with CO2, disappearance of the cavity is rapid and drainage is not needed. Moreover, this technique implies no restrictions on postoperative motility, and is thus suitable for elderly patients. PMID- 7482238 TI - Chronic subdural hematomas and seizures: the role of prophylactic anticonvulsive medication. AB - BACKGROUND: The rates of seizure activity associated with surgical treatment of chronic subdural hematoma (CSH) reported in the literature has varied greatly. The efficacy of prophylactic anticonvulsive medication (ACM) has been debated and its use been erratic. With improved diagnosis, reduction of associated morbidity impacts greatly on the mortality rate, and so the use of ACM may be important to the patient with CSH. METHODS: We conducted a retrospective analysis of the records of patients treated surgically for CSH and examined the prevalence of seizure activity, morbidity, and mortality and the effects of anticonvulsant medication. RESULTS: Ninety-eight patients (65 = male) were identified with a mean age of 72 years and a range of 4 to 97 years. Six patients had a preexisting seizure disorder; despite therapeutic serum levels of ACM, 3 of these 6 patients experienced seizures without mortality. Seizures occurred in 20/98 (20.4%) patients during hospitalization or follow-up. Of the 92 patients without a preexisting seizure disorder, 42 (46%) received prophylactic phenytoin with therapeutic serum levels. One patient among the 42 (2.4%) who received prophylactic ACM experienced seizure activity in comparison to 16 of 50 (32%) patients who did not receive adequate prophylactic ACM (P = < 0.001). Six of 11 deaths within one month of discharge occurred in patients with a new onset of seizures (P = < 0.005). Age, sex, history of trauma, Markwalder scores on admission, location of hematoma, or type of surgery were unrelated to the occurrence of seizures or mortality. Following hospital discharge, patients were continued on ACM for a mean duration of 8 months without seizures. CONCLUSIONS: The onset of new seizures was found in 17 (18.5%) of 92 patients and was associated with increases in morbidity and mortality. Patients who received prophylactic ACM demonstrated a significant decrease in the occurrence of seizures, and we therefore recommend the use of phenytoin prophylaxis in patients treated surgically for chronic subdural hematoma. PMID- 7482239 TI - Repeated decompressive craniectomy after head injury in children: two successful cases as result of improved neuromonitoring. AB - BACKGROUND Decompressive craniectomy in the treatment of posttraumatic brain swelling is not generally accepted. Until now the efficacy of operative decompressive craniectomy in posttraumatic brain swelling of children appeared more promising. However, the criteria for such procedures remain unclearly defined. METHODS We present two children who had repeated decompressive craniectomy following head injury, in order to control intracranial pressure (ICP) sufficiently. Our indications for performing a decompressive craniectomy in the presence of conservatively uncontrollable raised ICP are: (1) Patient is between the ages of 3 and 35 years. (2) An initial Glasgow Coma Scale (GCS) ranging between 4 and 8. (3) Three criteria have to be fulfilled at the same time: The cerebral perfusion pressure (CPP) has to drop to values of less than 60 mm Hg. It is impossible to control the ICP values (up to 45 mm Hg) conservatively. The diastolic velocity of the transcranial doppler sonography (TCD) has to decrease until only a systolic flow pattern is obtained. (4) No other mass lesion should be detected on cranial computed tomography (CCT) that could account for the rise in pressure. In both cases we performed bifrontal decompressive craniectomies. RESULTS Both patients survived. Seven months after the accident, patient No. 1 was oriented and could walk on her own with a mild right-side hemiparesis. Patient No. 2 could attend school 12 months postinjury. Both patients developed hygromas after the craniectomy. A shunt operation, however, was not necessary. CONCLUSIONS ICP monitoring, together with CCT examination, simultaneous recording of TCD, and systemic parameters, will reveal a patient at risk at a time when impending damage due to uncontrollable ICP may still be prevented. The simultaneous assessment of cerebral blood flow by transcranial doppler (TCD), in this situation, proves most valuable. It improves the guidelines of patient selection for decompressive craniectomy, in the presence of conservatively uncontrollable ICP. PMID- 7482240 TI - Intracerebral ring-enhancing lesions in a patient with multiple sclerosis: a case report. AB - BACKGROUND Intracerebral cystic lesions can easily be identified by computed tomography (CT) and magnetic resonance imaging (MRI). Cystic gliomas, brain abscesses, or metastatic carcinoma are the usual underlying causes for this neuroradiologic finding. CASE PRESENTATION We report here the case of a patient with multiple sclerosis who has two intracerebral cystic lesions. CONCLUSIONS Central nervous system (CNS) tissue destruction, necrosis, and cyst formation are very unusual in multiple sclerosis. Diagnostic difficulty therefore arises when patients suspected of having multiple sclerosis have radiologic evidence of cystic CNS lesions. PMID- 7482241 TI - Factors influencing tumor cell traffic in the central nervous system. PMID- 7482242 TI - Clinical research in intracranial aneurysms: an exercise in naming things correctly. Presidential address to the Society of Neurological Surgeons, San Francisco, May 27, 1980. PMID- 7482243 TI - Diffusion and perfusion imaging for acute stroke. PMID- 7482244 TI - The patient's contract with the doctor. PMID- 7482245 TI - Changing careers. PMID- 7482246 TI - Hot flushes in men with pituitary adenoma. AB - BACKGROUND: Hot flushes are common in menopausal women and also in men made acutely hypogonadal after orchiectomy or testicular injury. It is, however, an unusual symptom in patients with hypogonadism secondary to pituitary tumors. METHODS: In evaluating the histories of men with hypogonadal state associated with nonfunctioning pituitary macroadenoma we were struck by the presence of hot flushes in four of them. RESULTS: All four of the patients were hypogonadal with sexual dysfunction preoperatively. All had low gonadotropins and low testosterone levels with varying degrees of panhypopituitarism. All had successful transsphenoidal removal of tumors. None had endocrine improvement following surgery. All patients had improvement in sexual function and the hot flushes with administration of testosterone postoperatively. CONCLUSIONS: Hot flushes are an uncommon presentation in men with pituitary adenoma. Perhaps the symptom will become more prominent if it is specifically questioned. We postulate that the cause of the flushing is related to nonsuppressed pulsatile secretion of gonadotropin-releasing hormone (GnRH) from the hypothalamus. PMID- 7482247 TI - The pathologic, surgical, and MR spectrum of Rathke cleft cysts. AB - BACKGROUND: Rathke cleft cysts (RCC) uncommonly attain large enough size to cause symptoms or warrant neurosurgical intervention, leading to difficulty in recognizing their diverse presentations. METHODS: We retrospectively reviewed the clinical, intraoperative, pathologic, and magnetic resonance (MR) imaging findings of 16 cases seen at our institution since 1987. RESULTS: The most unusual clinical presentation was pituitary apoplexy, although CSF-like, cholesterol containing, or hemorrhagic cyst contents most often led to intraoperative misinterpretation. Microscopic features of squamous metaplasia, lymphocytic infiltrates, tubuloracemose glands, or coexistent adenomas demonstrated the pathologic variations in these intrasellar and suprasellar cysts. The MR findings of an anteriorly displaced pituitary stalk, the presence of a posterior ledge, and T2 hypointensity were distinctive, when present. CONCLUSIONS: Rathke cleft cysts have varied clinical presentations and sometimes confusing intraoperative and pathologic appearances. We document a new pathognomonic MR finding in these cysts of a posterior ledge sign. Our surgical approach is to open and drain cyst contents, treat the residual cyst lining with instillation of absolute alcohol, and close the floor of the sella with a bone strut. PMID- 7482248 TI - Sella turcica fractures in a trauma series and their clinical significance. AB - BACKGROUND: Fractures in the region of the sella turcica (ST) are usually associated with severe blunt head trauma and may produce many complications. The true incidence is difficult to assess since there have been only about 30 cases reported in the literature. METHODS: A total of 470 computed tomography (CT) sections--with optimum bone window images of skull bone--of 1600 head trauma patients who were admitted between 1979 and 1992, inclusive, to the Neurosurgery Department of Gazi University Medical School were analyzed retrospectively. RESULTS: There were a total of 17 patients with a fracture of the ST, constituting an incidence of 3.6%, and there was a preponderance of children. CONCLUSIONS: Computed tomography was very helpful in assessing the extent of ST fractures. In our cases, these fractures were not associated with a high complication or death rate, although a cerebrospinal fluid fistula was present in 53%. PMID- 7482250 TI - Traumatic basilar impression: case report. AB - A very rare case of traumatic basilar impression is reported. The patient, a 57 year-old man, was hit on the head vertically in the parietal region. X ray of the cervical spine and computed tomography (CT) scans showed intracranial indentation of the atlas and the odontoid process with a depressed fracture around the foramen magnum. There are no previous reports about this type of fracture. PMID- 7482249 TI - Should meningiomas involving the cavernous sinus be totally resected? AB - BACKGROUND: The surgical strategy for meningiomas involving the cavernous sinus (CS) is still controversial because of the difficult surgical approach and the risks of injury to the cranial nerves or the internal carotid artery. We evaluated the efficacy of subtotal removal of meningiomas involving the CS following chemical embolization, preserving cranial nerve functions. METHODS: We analyzed the histology, embolization material, recurrence rate, and neurologic complications in 19 fresh cases with meningioma involving the CS, who received subtotal removal surgery and were followed over 1 year. Patients were 6 men and 13 women aged from 30 to 69 years (mean, 52.2 years) on admission. All patients received preoperative chemical embolization of the feeding arteries to reduce intraoperative bleeding. In the 11 earlier patients, we used estrogen (Est) as the embolization material, but changed to estrogen with 25% alcohol+polyvinyl acetate (Est+PVac) for the latter 8 patients. RESULTS: Fourteen patients were recurrence free after 1-16 (6.5 +/- 5.5) years. The performance status of the survivors was good. Five patients, including two cases of malignant meningioma, developed recurrence, with a mean recurrence-free period of 1.7 +/- 0.5 years. Patients who received embolization with Est showed a higher recurrence rate than Est+PVac. CONCLUSIONS: These results suggest that subtotal resection following Est+PVac embolization is a safe and effective strategy for this tumor. PMID- 7482251 TI - Trigeminal neuralgia caused by a cerebellopontine-angle lipoma: case report. AB - This report describes the surgical management of a rare trigeminal neuralgia caused by a cerebellopontine (CP)-angle lipoma. A 13-year-old girl presented with typical trigeminal neuralgia in the right mandibular area. Magnetic resonance imaging (MRI) disclosed a lipoma in the right CP angle. A suboccipital craniectomy revealed a lipoma around the 7th and 8th cranial nerves, but there was no apparent lipoma on the surface of the trigeminal nerve. Since the trigeminal nerve swelled only by infiltration of the lipoma, a partial rhizotomy of the involved sensory root was inevitable. The surgery relieved her neuralgia. She remained analgesic in the mandibular area. PMID- 7482252 TI - Neurinoma of the trigeminal root and atypical trigeminal neuralgia: case report and review of the literature. AB - BACKGROUND: Neurinomas of the trigeminal nerve are a rare entity: those located in the posterior fossa account for 20% of all cases. In the majority of cases, the clinical presentation begins with fifth cranial nerve involvement producing a constant pain associated with other cranial nerve palsy and cerebellar signs. METHODS: We report the clinical features, neuroradiological imagings, and management of a case of trigeminal neurinoma located in the cerebellopontine angle, arising from the Vth cranial nerve root, presenting with an atypical trigeminal neuralgia; moreover, we analyze similar cases reported in the literature and we discuss whether pain can be produced by a neurinoma that arises central to the ganglion. RESULTS: The tumor presented with an atypical trigeminal neuralgia characterized by constant trigeminal pain with paroxystical burns, hyperesthesia and hyperactive autonomic dysfunction. Neuroradiological examinations provided the best preoperative localization of this lesion, allowing better planning of the surgical approach, considering the large size of this tumor. A retromastoid incision and posterior fossa craniectomy approach was used, with complete excision of the tumor. The paraxysmal sharp pain and hyperesthesia disappeared completely, but the constant burning pain persisted although it was less intense. At an 8-month follow-up examination, the patient showed a progressive improvement of clinical symptoms and control magnetic resonance imaging (MRI) showed the complete removal of the neoplasm and the absence of residuals or recurrences. CONCLUSION: Although in a high percentage of cases of atypical trigeminal neuralgia a neurovascular conflict might be suspected, the review of the present case suggests that the hypothesis of a trigeminal neurinoma must be investigated both with adequate neuroradiological procedures and/or microsurgical exploration of the trigeminal root. PMID- 7482253 TI - Adult-onset presentation of Dandy-Walker variant in siblings. AB - BACKGROUND: The Dandy-Walker syndrome and Dandy-Walker variant usually present as isolated cases of hydrocephalus in pediatric patients. METHODS AND RESULTS: THis paper consists of a case report of the adult onset of symptoms in two sisters having Dandy-Walker variant. Such an occurrence has never before been reported in the medical literature. Both patients presented with headaches and progressive neurologic deficit. On computed tomography (CT scan) of the head, both were found to have hydrocephalus, with hypoplasia of the inferior vermis. Both patients were treated successfully with ventriculoperitoneal shunting. A third sister, with a similar history, elected not to undergo CT scanning or surgical treatment. CONCLUSIONS: Variants of the Dandy-Walker syndrome may occasionally present clinically in the adult age group. Such an occurrence in siblings is consistent with an underlying genetic etiology. PMID- 7482255 TI - Basis of brain tumor biology. PMID- 7482254 TI - Familial syringomyelia: case report and review of the literature. AB - BACKGROUND: Syringomyelia is an uncommon disease of the spinal cord, occurring sporadically. However, rare familial cases with autosomal dominant or recessive inheritance patterns are reported and their incidence quoted as approximately 2%. Only one previous report originated from the United States. METHODS: We present a brother and sister with syringomyelia and associated Chiari type I malformation; both patients responded to surgical treatment. We review the world literature and briefly discuss pathogenetic theories of syringomyelia as well as the relevance of the histocompatibility leukocyte antigen profile. RESULTS: Both genetic and environmental factors appear to be involved in familial syringomyelia. CONCLUSION: We recommend that close relatives of patients affected with familial syringomyelia undergo routine neurologic and radiologic surveys. PMID- 7482256 TI - Effect of mannitol on focal cerebral ischemia evaluated by somatosensory-evoked potentials and magnetic resonance imaging. AB - BACKGROUND: Mannitol has been used in routine neurosurgical practice for the control of increased intracranial pressure. The effect of mannitol on focal cerebral ischemia was evaluated by somatosensory-evoked potentials (SEP) and magnetic resonance imaging (MRI). METHODS: The left middle cerebral artery (MCA) was exposed via the superomedial transorbital approach and occluded proximal to the origin of the perforating arteries. Ten cats received mannitol (0.5 g/kg IV) immediately, 6, 12, and 18 hours after MCA occlusion. The other 10 cats received saline solution and served as control. The animals were initially prepared to measure SEP before and 15, 30, and 60 minutes after MCA occlusion. Following SEP measurement, all cats were prepared for MRI. Sequential MRI of both intravoxel incoherent motion (IVIM) and T2-weighted spin echo techniques were obtained at 2, 4, 6, and 24 hours after MCA occlusion. The animals were sacrificed after the last MRIs for histologic study. RESULTS: The SEP amplitude decreased to about 10% at 15 minutes after MCA occlusion and then gradually recovered to 38% at 60 minutes in the mannitol group, and 21% in the control group. In MRI study, IVIM imaging demonstrated ischemic cerebral injury as a sharply demarcated area at 2 hours after MCA occlusion, while T-2 weighted imaging failed to show clear evidence of injury until 2-6 hours. High-signal intensity areas on both IVIM and T2-weighted images were smaller in the mannitol group than those in the control group. Histologic study demonstrated that infarction size was 36.9% +/- 7.7% of the left hemisphere in the mannitol group and 57.3% +/- 5.3% in the control group (p < 0.05). CONCLUSIONS: Mannitol is effective for acute cerebral ischemia, and SEP and MRI are useful for monitoring it. PMID- 7482257 TI - Hemodynamic evaluation with dynamic DSA during the treatment of cerebral vasospasm: a retrospective study. AB - BACKGROUND: Percutaneous transluminal angioplasty (PTA) and super-selective intra arterial infusion of papaverine have recently been performed for the treatment of cerebral arterial vasospasm, with generally favorable results in patients with delayed symptomatic cerebral vasospasm. METHODS: We used dynamic digital subtraction angiography (dynamic DSA) to measure local transit time before and after endovascular treatment (EVT) for the treatment of ruptured intracranial aneurysm and delayed symptomatic cerebral vasospasm. Mean transit time (MTT) was measured by dynamic DSA in 19 patients with ruptured intracranial aneurysm and delayed symptomatic cerebral vasospasm and 6 patients without symptomatic cerebral vasospasm (group III) just after surgery, immediately prior to EVT, just after EVT, and in the chronic stage of disease more than 1 month after onset of subarachnoid hemorrhage. EVT included PTA using a silicone nondetachable balloon and/or superselective infusion of 0.2% papaverine. RESULTS: Symptomatic vasospasm occurred between days 7 and 11 after onset of subarachnoid hemorrhage. The MTT just after the onset of neurologic deterioration in these 10 patients with complete neurologic recovery during the chronic stage of disease (group I) and the other 9 patients without complete recovery (group II) were respectively 6.92 +/- 0.42 seconds and 7.66 +/- 0.78 seconds. The latter value was significantly larger than the former (p < 0.02). MTT just after EVT in group II were also significantly greater than the corresponding MTT in group I (p < 0.002). In addition, immediately prior to EVT the MTT in group I and group II was significantly longer than that in group III (p < 0.0001). CONCLUSIONS: Functional analysis with dynamic DSA can be used to detect hemodynamic disturbances, and can be used for hemodynamic evaluation during the treatment of symptomatic cerebral vasospasm. PMID- 7482258 TI - Angiographic workup of a carotid cavernous sinus fistula (CCF) or what information does the interventionalist need for treatment? . AB - It is rare that a patient with a carotid cavernous sinus fistula (CCF) is referred to the interventionalist with a good and exhaustive angiographic workup, providing all the information necessary for a therapeutic decision. The goal of this technical note is to refresh our memory on the best way to study this group of patients. It does not have the pretension to bring a new technique. As there are two types of CCFs, direct and indirect, there are also two different angiographic workups. PMID- 7482259 TI - Neurosurgical management of the frontal sinus. PMID- 7482260 TI - Acromegaly and pituitary tumors: early anatomoclinical observations. AB - The elucidation of pituitary physiopathology has been a major challenge for physicians since early ages. Due to the unawareness of the endocrine system, acromegaly, the most striking pituitary disorder, was commonly regarded as an intrinsic bone disease. Andrea Verga and Vincenzo Brigidi, Italian authors of the 19th century, reported the first macroscopic and microscopic descriptions of pituitary adenomas in acromegalic patients. Although far from providing a correct pathogenetic interpretation of the disease, they opened the way to forthcoming observations and discoveries. A short history of acromegaly and pituitary physiopathology is drawn. PMID- 7482261 TI - Publication world and world publications. PMID- 7482262 TI - Use of outer layer of dura as a pedicled flap. PMID- 7482263 TI - Facial nerve outcome after acoustic neuroma surgery. PMID- 7482264 TI - Rectal squamous mucosectomy and ileal anal pull-through procedures: single surgeon experience in 105 patients. AB - BACKGROUND: The ileal anal pull-through procedure has become the most frequently used cointinence sparing procedure for patients with ulcerative colitis or familial polyposis. Areas of controversy concern the use of temporary ileostomies, and the extent of the rectal mucosectomy. The current report presents a single surgeon's experience with mucosectomy to the perianal skin (squamous mucosectomy), with ileal J-pouch reservoir construction and temporary ileostomy. METHODS: We reviewed the records of 105 consecutive patients undergoing this procedure by a single surgeon during a 5-year period. One hundred percent follow-up was achieved. RESULTS: There was 100% gross fecal continence, with 5% of patients expressing rare day time leakage, and 28% having intermittent nocturnal leakage. There were no instances of pelvic sepsis, and no pouches have been removed. The diverting ileostomy was associated with 6% morbidity. CONCLUSIONS: We conclude that the rectal mucosectomy can be safely extended to the levels of perianal skin with no loss in continence or function. We recommend that this be adopted as the standard for this procedure to ensure complete eradication of the underlying pathologic condition. PMID- 7482265 TI - Factors associated with a positive reexcision after excisional biopsy for invasive breast cancer. AB - BACKGROUND: Breast-conserving therapy followed by adjuvant radiotherapy represents an alternative to mastectomy as a treatment for invasive breast cancer. When excisional biopsy has been performed outside the parent institution, reexcision is often performed, with tumor being identified in 32% to 62% of the subsequent specimens. We analyzed not only the factors associated with a positive reexcision but also those factors associated with final surgical margins that are positive for tumor. METHODS: Between 1978 and 1991, 956 female patients with American Joint Committee on Cancer clinical stage I or II breast cancer were treated with breast-conserving therapy where a total of 420 patients underwent reexcision after an initial excisional biopsy. Several factors were analyzed to determine their association with a positive reexcision, the status of the final surgical margin, and the nature of the disease present within the reexcision specimen. RESULTS: Factors that correlated with a positive reexcision in both univariate and multivariate analysis were clinical tumor size, method of detection, the pathologic status of the axillary lymph nodes, and the histologic appearance. Those factors associated with finding invasive disease at the time of reexcision were clinical tumor size, clinical presentation, and nodal status. The single factor associated with finding residual in situ disease at the time of reexcision was histologic appearance of the primary tumor. A final positive margin was associated with method of tumor detection, age of the patient, and the presence of axillary lymph node metastases. CONCLUSIONS: The most significant factors associated with a positive reexcision are clinical tumor size, method of tumor detection, pathologic nodal status, and histologic appearance. Patients with larger tumors or those that are detected by physical examination, as well as invasive lobular carcinomas, may require a more generous initial resection to achieve negative surgical margins and avoid the likelihood of reexcision. PMID- 7482266 TI - Economic implications of thrombolysis or operation as the initial treatment modality in acute peripheral arterial occlusion. AB - BACKGROUND: Thrombolytic agents have been used to treat arterial occlusion for almost 40 years. Recently, an investigation of the costs associated with two treatment options for acute peripheral arterial occlusions, thrombolysis and surgical intervention, was completed. The availability of hospital cost data for patients enrolled in the thrombolytic and operative treatment groups provided a basis with which to accurately assess the financial impact of the different treatment strategies, both from a purely financial standpoint and in relation to outcome. METHODS: The patient base was composed of 114 patients with acute limb ischemia of less than 7 days' duration. The patients were randomly assigned to receive urokinase (n = 57) or to undergo an operation (n = 57) as the initial therapeutic intervention. Patients in the thrombolytic group underwent standard intraarterial diagnostic arteriography, and patients with embolic events, in whom complete lysis of all embolic and propagated thrombotic material was achieved, were subsequently treated with heparin and long-term warfarin (Coumadin) therapy. The economic analysis was undertaken after the completion of the trial. Statistical comparisons between groups were made with the Student t test for continuous, normally distributed data. Mortality and limb salvage rates were calculated from Kaplan-Meier curves, appropriate for the censored nature of the data. RESULTS: The total treatment costs did not differ significantly between the two treatment groups ($22,171 +/- $4,959 in the thrombolytic group and $19,775 +/ $5,253 in the operative group). The total hospital charges were similar between the two groups. Overall, the total charges were remarkably similar between the two treatment groups, averaging $40,823 +/- $8,764 in the thrombolytic group and $41,930 +/- $10,398 in the operative group. CONCLUSIONS: An economic analysis of the data confirmed that the total economic impact of thrombolysis approximated that of initial operative therapy. The improved clinical outcome in patients treated with thrombolysis suggests that this modality may be appropriate as the initial therapeutic intervention in the select group of patients seen within the first few hours of an acute peripheral arterial occlusive event. PMID- 7482267 TI - Early closure of colostomies in trauma patients--a prospective randomized trial. AB - BACKGROUND: Most traumatic colon injuries can be repaired primarily, but a colostomy may still be required for severe colonic or rectal injury. The current trend is to reverse the colostomy early, rather than to wait the traditional 3 months before closure. METHODS: Forty-nine patients with colostomies after abdominal trauma were entered into the study. All patients had undergone a contrast enema in the second postoperative week to assess distal colon healing. Patients were excluded from early closure for nonhealing of the bowel injury, unresolving wound sepsis, or an unstable condition. We then compared the outcome of the remaining 38 (77.6%) patients allocated to either an early or a late colostomy group in a controlled, prospective, randomized trial. RESULTS: We found no significant difference in morbidity between the two groups, with an overall complication rate of 26.3%. Technically the early closure of colostomies was far easier than late closure and required significantly less operating time (p = 0.036) and with less intraoperative blood loss (p = 0.020). The closure of end colostomies was more time consuming, both early (p < 0.001) and late (p < 0.001) and caused more bleeding (p < 0.001 and p < 0.001, respectively). Total hospitalization was marginally shorter overall for early closure, but late closure of end colostomies resulted in prolonged hospitalization (p = 0.023). CONCLUSIONS: The early closure of colostomies and the use of loop colostomies whenever possible are recommended as both safe and beneficial for patients with colonic injury after trauma. Contraindications for early closure include nonhealing distal bowel, persistent wound sepsis, or persistent postoperative instability. PMID- 7482268 TI - Papillary cystic and solid tumors of the pancreas: a pancreatic embryonic tumor? Studies of three cases and cumulative review of the world's literature. AB - BACKGROUND: The papillary cystic and solid tumor of the pancreas is rare. It occurs predominantly in young women, and most present a benign behavior. The pathogenesis of this tumor has attracted a number of investigations but remains unclear. METHODS: We present three patients with this tumor and a review of 289 others from the world's literature, a total of 292 cases. On the basis of the analyses of the clinical and pathologic features from the reported cases, the pathogenesis of this unusual tumor has been further explored. RESULTS: Ninety percent of the patients were female, with a mean age of 23.9 years. The tumors were usually quite large with a mean diameter of 10.3 cm. Ninety-two percent of these tumors were totally or partially cystic. Rupture of the capsule resulted in hemoperitoneum in eight cases, five of which were without any identifiable cause. Forty-three tumors (14.7%) have been recognized as malignant. The overall prognosis has been excellent and an aggressive approach to resection is indicated. CONCLUSIONS: The results of immunohistochemical staining and electromicroscopy were rather diverse, but most, including the current cases, support the hypothesis that the tumor originates from pleuripotential embryonic stem cells. Thus the term pancreatic embryonic tumors seems preferable to papillary cystic and solid tumor of the pancreas to delineate the origin of the tumor and to reflect some of its biologic characteristics. PMID- 7482269 TI - Nerve injury associated with laparoscopic inguinal herniorrhaphy. AB - BACKGROUND: As laparoscopic herniorrhaphy becomes more popular, it is important to realize the potential for injury to surrounding neural structures, with attendant severe disability. METHODS: Herein are discussed two patients with disabling neuralgia after laparoscopic herniorrhaphy. RESULTS: Both patients were treated with transabdominal removal of their prosthetic materials and anchoring staples, with dramatic symptomatic improvement. CONCLUSIONS: The surgeon should be aware of the anatomic considerations accompanying laparoscopic herniorrhaphy. In regard to nerve injury, laparoscopic herniorrhaphy may pose certain disadvantages over traditional hernia repairs. It may diminish the ability to appreciate the course of nerves in the inguinal region and their relationship to the spermatic cord, and injury to nerves may be difficult to recognize and treat. PMID- 7482270 TI - Age-related differences in the distribution of peripheral atherosclerosis: when is atherosclerosis truly premature? AB - BACKGROUND: The natural history of peripheral atherosclerosis in young adults appears to be unfavorable compared with that in older patients. No universally accepted definition of "premature" atherosclerosis exists, however, making comparison of clinical studies difficult. This study examined age-related differences in distribution of atherosclerotic lesions and determined an age threshold at which such differences became apparent. Such a threshold may provide a definition of premature atherosclerosis. METHODS: Arteriograms of all patients 49 years of age and younger undergoing evaluation of lower extremity ischemia during the past 5 years were reviewed and the findings were tabulated. Medical records were reviewed to obtain demographic data, assess risk factors, and confirm disease etiology. Exclusion criteria included normal arteriograms (three patients), history of acute or remote trauma (six patients), unclear cause of ischemic symptoms (three patients), arteritis (four patients), aneurysmal disease (one patient), and acute ischemia without prior chronic symptoms (12 patients). For comparison we also reviewed arteriograms performed during the same period in 140 patients older than 50 years of age who had chronic lower extremity ischemia caused by atherosclerosis. RESULTS: The mean age of the 59 study patients was 43.4 +/- 5.8 years (median age, 46 years; range, 25 to 49 years). Arteriograms were available in all cases; medical records were available in 54 (92%). Atherosclerosis involved only the aortoiliac segment in 25 patients (42%), the femoropopliteal-tibial arteries alone in 21 (36%), and both levels in 13 (22%). Patients with distal atherosclerosis had a higher prevalence of diabetes than those with proximal atherosclerosis (p = 0.004). Ninety-two (66%) of the 140 patients older than 50 years of age had atherosclerosis confined to a single arterial segment, which was not significantly different from the prevalence of single-level disease in the study group. However, 25 (54%) of the 46 study patients with single-level atherosclerosis had aortoiliac disease compared with only 15 (16%) of 92 patients older than 50 years of age with single-level disease (p < 0.001). CONCLUSIONS: In contrast to the pattern of disease in older adults, atherosclerosis in young, nondiabetic patients most commonly involves the aortoiliac segment. Differences in lesion distribution become increasingly apparent with age but are most striking between those 49 years of age and younger and those 50 years of age and older. Accordingly, we propose that premature peripheral atherosclerosis be defined as beginning at or before the age of 49 years. PMID- 7482271 TI - Long-term outcome after transaxillary approach for thoracic outlet syndrome. AB - BACKGROUND: Recurrence or persistence of neurologic symptoms after surgical treatment of patients with thoracic outlet syndrome (TOS) are reported to be as high as 25%. To identify factors affecting the long-term outcome of surgical treatment of patients with TOS, we reviewed our 20-year experience. METHODS: One hundred thirty-four transaxillary first rib resections were performed on 118 patients (43 men, 75 women, mean age 38 +/- 13 years). Eighty-three operations (61.9%) were undertaken to relieve symptoms resulting from compression of the lower roots of the brachial plexus, 37 (27.6%) for compression of both lower and upper roots, and 14 (10.5%) for lower root and vascular symptoms. All patients underwent a transaxillary extraperiosteal first rib resection with transection of the scalene muscles. In 73 cases (54.5%) a resection of the anterior scalene muscle was also performed. A cervical rib was removed in 28 cases (20.1%), and anomalous fibrous bands adjacent to the neurovascular bundle were resected in 41 cases (30.6%). RESULTS: No major complications were observed. Of 105 patients (118 procedures) followed up (mean follow-up, 99 +/- 72 months), good to excellent results were obtained in 96 cases (81.4%) and fair to poor results were recorded in 22 cases (18.6%). The presence of a long posterior first rib stump, measured from the chest x ray films, was the strongest determinant of the long term results among the variables examined (p < 0.0001). Reoperation, consisting of neurolysis and resection of the stump, was performed in 16 patients. The results were excellent in all cases at a mean follow-up of 66 +/- 46 months. Primary and secondary 10-year, actuarial freedom rates from recurrent symptoms were 80.9% and 93.1%, respectively. CONCLUSIONS: Our results suggest that the long-term outcome after surgery for TOS was strongly influenced by the extent of the first rib resection. PMID- 7482272 TI - Prognostic factors of resected adenocarcinoma of the esophagus. AB - BACKGROUND: The main purpose of this study was to determine prognostic factors in patients with surgical treatment of adenocarcinoma of the esophagus. METHODS: Within a 12.5-year period, esophageal adenocarcinoma was resected in 165 patients by radical transhiatal esophagectomy (n = 134) or transthoracic en bloc esophagectomy (n = 31). Tumors were analyzed according to the 1992 UICC classification with respect to pTNM stage, residual tumor (R) status, grading, and ratio of infiltrated to resected lymph nodes (lymph node ratio); both univariate and multivariate analysis of prognostic factors were performed. RESULTS: The 30-day mortality rate was 6.1%. A complete removal of the tumor was achieved in 83% of the patients. Lymph node metastases were not detected in mucosal cancer (pT1a) but were detected in 18% of submucosal cancer (pT1b), 77% of pT2, 83% of pT3, and 96% of pT4. The overall 5-year survival rate was 34%; for patients without postoperative residual tumor (R0) it was 41%, and for those without lymph node metastases (pN0, R0) 63%. The 5-year survival rate for patients (pN1) with less than 30% invaded lymph nodes was 45%, compared with 0% for more than 30% invaded nodes. Independent prognostic factors for R0 resected patients excluding postoperative fatal outcome were pT and lymph node ratio. CONCLUSIONS: Long-term survival after resection of esophageal adenocarcinoma is mainly associated with complete tumor removal, limited esophageal wall penetration, and ratio of infiltrated to removed lymph nodes of less than 0.3. PMID- 7482273 TI - Surgical management of subclavian vein obstruction, including six cases of subclavian vein bypass. AB - BACKGROUND: The indications are still being developed for the various procedures available to treat thrombotic and nonthrombotic subclavian vein obstruction. This article explores our experience with the surgical options, primarily with chronic venous occlusion, but also with acute thrombosis. METHODS: Twelve patients presented with acute or chronic venous thrombosis or nonthrombotic venous obstruction. Treatment included thrombolysis, vein patch angioplasty, thrombectomy, first rib resection, venolysis, and/or subclavian vein bypass by either jugular vein transposition or prosthetic axillojugular bypass. RESULTS: Six patients experienced relief of all symptoms; the other six have had relief from pain but continue to have occasional swelling. CONCLUSIONS: On the basis of our anecdotal experiences and those of others, the following protocol has been developed. After acute thrombosis, lytic therapy is recommended. If successful, first rib resection and venolysis should be considered; if unsuccessful and the patient has significant symptoms, immediate thrombectomy is indicated. If patency is established, rib resection and venolysis should follow. Persistent stenosis is treated by angioplasty, either with a vein patch or percutaneously. Complete occlusion is managed by jugular vein transposition or axillojugular bypass. Temporary arteriovenous fistulas accompany all open venous repairs. PMID- 7482275 TI - Necrotizing fasciitis: an indication for hyperbaric oxygenation therapy? AB - BACKGROUND: The accepted treatment protocol for necrotizing fasciitis (NF) consists of extensive surgery and wide spectrum antibiotics. Hyperbaric oxygenation (HBO) has been recommended as adjuvant therapy for NF, improving patient mortality and outcome. However, the beneficial effect of HBO for NF remains controversial. METHODS: A retrospective evaluation of treatment outcome in 37 patients treated for NF between 1984 and 1993 was carried out. The mortality rate, morbidity criteria, and risk factors for grave prognosis were compared between a group of 25 patients who received HBO as part of their treatment protocol and a group of the remaining 12 patients treated by surgical excision and antibiotics alone. RESULTS: The two groups were found to be similar with regard to age, gender, the incidence of individual risk factors for ominous prognosis, and the Acute Physiology and Chronic Health Evaluation (APACHE) II score for disease's severity on presentation. The mortality rate among the HBO treated patients was 36%, as opposed to 25% in the non-HBO group. The mean number of surgical debridements required per patient was significantly higher in the HBO group: 3.3 compared with 1.5 in the non-HBO-treated patients. Although the average length of hospitalization for survivors was shorter for the HBO group, the difference between the groups did not reach statistical significance. CONCLUSIONS: The results of this study cast doubt on the suggested advantage of HBO in reducing patient mortality and morbidity when used as adjuvant therapy for NF. PMID- 7482274 TI - Correlation between cystoscopic biopsy results and hypoamylasuria in bladder drained pancreas transplants. AB - BACKGROUND: Urinary amylase (UA) remains the most common biochemical parameter to detect rejection in bladder-drained pancreas allografts. With the development of the cystoscopic transduodenal pancreas transplant biopsy technique, tissue samples of the pancreas graft are now frequently obtained. A definitive correlative analysis between UA activity and biopsy results has not been done in the three different pancreas transplant categories (simultaneous pancreas-kidney, pancreas transplant alone, and pancreas after kidney). METHODS: We studied 66 pancreaticoduodenal biopsy specimens obtained for hypoamylasuria. Rejection was defined as a greater than 25% decrease from stable posttransplantation baseline on two consecutive measurements at least 12 hours apart. To perform biopsies we used our newly developed 14- and 16-gauge core-cut needles (50 cm long). Biopsy specimens were considered positive if either pancreatic or duodenal rejection was found. To assess the quality of UA activity we studied 13 biopsy specimens from patients with stable UA levels; these 13 specimens were negative for rejection. RESULTS: Acute rejection was diagnosed in 36 biopsy specimens (55%). The mean decrease in UA levels was 67% +/- 8% (range, 28% to 99%) for the positive biopsy results, and 57% +/- 16% (range, 22% to 92%) for the negative biopsy results (p = 0.147). Within 1 month, UA levels returned to baseline in 19% of our patients with positive biopsy results versus 97% with negative results; postbiopsy 1-year graft survival was 64% versus 97% (p < or = 0.05). In assessing the test quality of our biopsy specimens (including 13 obtained for reasons other than hypoamylasuria), we found a sensitivity of 100% (stable UA levels mean no rejection) and a specificity of 30%. The predictive value of a positive test was 53%; of a negative test it was 100%. By performing biopsies we avoided antirejection treatment in 47% of the patients studied. We found no biopsy related complications. CONCLUSIONS: Stable UA levels reliably rule out rejection; a decrease is a marker for acute rejection but is unspecific. Performing biopsy is currently the only way to reliably diagnose rejection in solitary pancreas recipients (pancreas transplant alone and pancreas after kidney) and in simultaneous pancreas-kidney recipients with isolated hypoamylasuria. The procedure is safe and should always be attempted to avoid unnecessary rejection treatment. PMID- 7482276 TI - Bronchoscopic guidance makes percutaneous tracheostomy a safe, cost-effective, and easy-to-teach procedure. AB - BACKGROUND: We wanted to assess the efficiency of instituting a modified technique of percutaneous tracheostomy (PET) with bronchoscopic guidance. METHODS: During a 10-month period 48 consecutive trauma patients requiring tracheostomy were divided between a standard tracheostomy control group (ST) and a PET group. All patients were followed prospectively. The hospital charges were reviewed retrospectively. RESULTS: Age, gender, body habitus, and principal diagnosis were similar in the 21 ST patients and the 27 PET patients. All STs and 15 of the PETs were performed in the operating room (OR), and the 12 remaining PETs were done in the intensive care unit (ICU). Four patients in the ST group and six in the PET group died. One of these deaths occurred in a patient in the PET group with severe adult respiratory distress syndrome. Procedure time was shorter for PET (16 versus 45 minutes, p < 0.0001). Junior residents performed more PETs than STs (33% versus 10%), and PET was considered "easier" to perform than ST (81% versus 47%). Hospital charges for PET in the ICU were $3400 less per patient compared with ST or PET in the OR. CONCLUSIONS: PET was performed easily and safely in the OR and at the ICU bedside. PET required one-third the time of ST. Bronchoscopic supervision of PET may have contributed to the small number of complications and the educational experience of junior residents. PET in the ICU can reduce hospital charges significantly and avoids transport of patients to the OR. PET is as safe as ST and should be considered the procedure of choice for an ICU patient requiring an elective tracheostomy. PMID- 7482277 TI - Effects of early and delayed wound excision on pulmonary leukosequestration and neutrophil respiratory burst activity in burned mice. AB - BACKGROUND: Tissue myeloperoxidase (MPO) is a marker of neutrophil (PMN) accumulation in tissues (leukosequestration). We measured MPO in the livers, guts, and lungs of mice after burn injury and studied the additive effect of burn excision on lung MPO. Lung histologic characteristics were also examined. PMN respiratory activity was assessed by measuring intracellular H2O2 content. METHODS: Mice received 32% total body surface area (TBSA) burns; some underwent burn excision followed by wound closure with allograft skin, either immediately or 48 hours after burn. Tissue MPO was measured by a colormetric assay, and intracellular H2O2 was quantified by flow cytometry. RESULTS: MPO was elevated in lungs 8 to 24 hours after burn (p < 0.05) but not in the liver or ileum. Other burned mice received either immediate or 48-hour-delayed wound excision and allografting. In controls a similar-size area was excised and grafted with normal or burned skin. Burned animals had increased lung MPO compared with nonburned animals (p < 0.05). Highest lung MPO levels were seen after burn/immediate excision (p < 0.001). Lung MPO levels were not different comparing unburned mice undergoing skin excision and grafting with either nonburned or burned skin. When burn excision was delayed 48 hours, lung MPO was increased moderately (p < 0.05) but remained far below levels in mice that were excised immediately after burn. PMN influx into lung tissues was confirmed by histologic examination. PMN H2O2 production was increased in burned mice and was additionally increased after immediate wound excision. CONCLUSIONS: Although burn injury produces pulmonary leukosequestration, the phenomenon is unrelated to local effects of burned skin. In this experimental model immediate postburn wound excision increased pulmonary leukosequestration to higher levels than after burn injury alone, and intracellular H2O2 content also increased. Pulmonary leukosequestration may predispose to lung injury, possibly limiting the benefits of wound excision performed extremely early postburn. PMID- 7482279 TI - Localized giant cell tumor of tendon sheath of upper back. PMID- 7482278 TI - Scintigraphy and biodistribution of monoclonal adrenocortical antibody in mice grafted with human adrenocortical carcinoma. AB - BACKGROUND: A murine monoclonal antibody recognizing normal and neoplastic human adrenocortical cells has been evaluated for scintigraphic localization and biodistribution in 53 nude mice grafted subcutaneously with human adrenocortical cell lines SW-13 and T-CAR 1. METHODS: The immunoglobulin G1 antibody and its Fab'2 fragment were purified and labeled with 125I. The tumor grafts exhibited diameters of 5 to 15 mm at 4 to 6 weeks after transplantation, when mice received a single subcutaneous or intraperitoneal injection of 50 micrograms iodinated intact or fragmented antibody, respectively. RESULTS: Examination up to 8 days after immunoglobulin G administration showed mean radioactivity ratios less than 1.0 for tumor to blood and corresponding ratios in tumor to lung, liver, spleen, and kidney from 0.6 to 5.3 at the time of peak tumor to blood ratio. A high background activity was noted on scintigraphic tumor visualization with the iodinated immunoglobulin G. In contrast, the radiolabeled Fab'2 fragment displayed gradually rising tumor to blood ratios, which, 4 days after injection, averaged 10.5 for T-CAR1 and 5.3 for SW-13. Tumor transplants were scintigraphically visualized without substantial background activity 3 days after Fab'2 injection, when the ratio of radioactivity in the tumor to the investigated murine organs was 0.5 to 7.3. CONCLUSIONS: The findings substantiate that immunoscintigraphy with the Fab'2 fragment of the antiadrenocortical Ac5 antibody may become a tool to localize human adrenocortical carcinoma. PMID- 7482280 TI - Monosegmental liver transplantation. PMID- 7482281 TI - Successful repair of left ventricular rupture caused by blunt trauma: a case report. PMID- 7482283 TI - Is it time to change the prothrombin time? PMID- 7482282 TI - Repeated radical resection and intraoperative irradiation for recurrent pancreatic ductal adenocarcinoma after pancreatoduodenectomy. PMID- 7482284 TI - Infraperitoneal colorectal anastomosis: hand-sewn versus circular staples. PMID- 7482285 TI - Availability of specific endotoxin antagonists. PMID- 7482287 TI - Long-term results after operation for pancreatic pseudocysts. PMID- 7482286 TI - Is there a last word in inguinal herniorrhaphy with prosthesis? PMID- 7482288 TI - Effects of local connectivity on striatal function: stimulation and analysis of a model. AB - Neuronal population activity was investigated by computer simulation of a network model based on the neostriatum. Three network topologies were studied, based on different assumptions about the synaptic connectivity among medium spiny neurons. In all networks neurons were interconnected by inhibitory synapses. The connectivity was either symmetric, in which case all connections between cells were reciprocal and equal in strength; or asymmetric. Simulations showed that networks with symmetric connectivity receiving randomly distributed afferent excitation produced stationary spatial activity patterns. In contrast, asymmetric connectivity in homogeneous networks produced slow travelling-wave activity across the network. We suggest that the shape of the medium spiny neurons is an important determinant of synaptic connectivity and that changes in the shape of these neurons caused by Huntington's disease would result in asymmetric connectivity. Slow travelling-wave activity produced by asymmetric connectivity in the neostriatum could explain some aspects of the choreic movement and some electromyographic features seen in Huntington's patients. PMID- 7482289 TI - Corticosteroid-mediated modulation of carbachol responsiveness in CA1 pyramidal neurons: a voltage clamp analysis. AB - Pyramidal neurons in the rat hippocampal CA1 area contain mineralocorticoid (MRs) and glucocorticoid (GRs) receptors for corticosterone. Previous current clamp experiments showed that depolarizations evoked by carbachol (1-3 microM) depend on relative MR/GR occupation: carbachol responses are small with predominant MR activation and larger when both receptor types are occupied. Multiple K conductances underlie the carbachol-induced depolarization. In the present study we used the single electrode voltage clamp technique to examine which K conductances modulated by carbachol are sensitive to corticosteroid treatment in vitro. We observed that 1 microM carbachol significantly reduced the IK,Leak while the IM was hardly affected; carbachol effects on the IK,Leak were significantly reduced under conditions of predominant MR activation compared to simultaneous activation of MRs and GRs. With a higher (10 microM) carbachol dose, steroid modulation of the IK,Leak showed a similar tendency. The amplitude of the IM was largely reduced by 10 microM carbachol but appeared to be not affected by steroid treatment. We conclude that the previously described suppression of the carbachol-induced depolarization with predominant activation MRs is caused by an attenuation of the carbachol action on the IK,Leak. PMID- 7482290 TI - Time-dependent changes in dopamine agonist-induced striatal Fos immunoreactivity are related to sensory neglect and its recovery after unilateral prefrontal cortex injury. AB - This study examined interactions between the corticostriatal glutamatergic system and the nigrostriatal dopaminergic system via immunocytochemical examination of dopamine (DA) agonist induction of the striatal immediate early gene product Fos following cortical injury. After unilateral aspiration of the medial agranular cortex (AGm) region of prefrontal cortex, rats were tested for orientation to visual, tactile, and auditory stimuli. Fos immunoreactivity induced by d amphetamine (5 mg/kg) or apomorphine (5 mg/kg) was quantified in dorsolateral and ventrolateral regions of caudate-putamen (CPu) in rats still demonstrating sensory neglect (5 days postsurgery) and in rats recovered from sensory neglect produced by AGm ablation (29+ days postsurgery). The pattern of immunoreactivity of rats still demonstrating neglect differed from that of unlesioned rats or recovered AGm-ablated rats. In rats demonstrating sensory neglect, d-amphetamine or apomorphine induction of Fos in the ipsilateral CPu was reduced by about 40% compared to the contralateral CPu or to comparable readings in unlesioned controls. These asymmetries were restricted to dorsolateral CPu, the region receiving the densest input from AGm. In contrast, recovered AGm-ablated rats had DA agonist-induced striatal Fos immunoreactivity that was symmetrical between the two hemispheres and comparable to control values. These findings indicate that adaptations involving the striatal medium spiny neuron, a site of convergence of cortical glutamatergic and nigral dopaminergic afferents, may contribute to recovery from behavioral deficits resulting from neocortical injury. PMID- 7482291 TI - IPT: a novel iodinated ligand for the CNS dopamine transporter. AB - An iodinated cocaine derivative, N-(3'-iodopropen-2'-yl)-2 beta-carbomethoxy-3 beta-(4-chlorophenyl) tropane (IPT), was evaluated as a probe for in vitro and in vivo labeling of dopamine (DA) and serotonin (5-HT) transporters in Sprague Dawley rat brain. Saturation analysis of [125I]IPT in rat striatal homogenates, in two different buffer solutions, Tris-HCl and phosphate, demonstrated a one site binding with affinities (Kd) of 0.25 +/- 0.02 and 0.16 +/- 0.02 nM and densities (Bmax) of 939 +/- 161 and 1,982 +/- 137 fmol/mg protein, respectively. Competition by known DA transporter ligands showed a rank order of RTI-55 > IPT > GBR12909 > mazindol > (-)cocaine. Binding to 5-HT transporter sites was evaluated in rat cortical homogenates. Saturation experiment results showed a single site with a Kd value of 1.2 +/- 0.2 nM and a Bmax value of 100 +/- 20 fmol/mg protein. The rank order of potency of several monoamine uptake inhibitors (paroxetine > fluoxetine > mazindol > R-nisoxetine > GBR12909) suggests that [125I] IPT labels 5-HT transporters in rat cortical homogenates. Both ex vivo and in vitro autoradiographic studies revealed high densities of [125I]IPT binding sites in the caudate nucleus, putamen, olfactory tubercle and nucleus accumbens, areas known to be rich in dopaminergic innervation. Moderate accumulation of activity was also observed in the substantia nigra. The dorsal raphe, a region with a high density of 5-HT innervation, was labeled using in vitro autoradiography with [125I]IPT, but the labeling using ex vivo autoradiography was less prominent at 30 min postinjection and not noticeable at 60 min postinjection.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7482293 TI - In vivo characterization of two cell types in the rat globus pallidus which have opposite responses to dopamine receptor stimulation: comparison of electrophysiological properties and responses to apomorphine, dizocilpine, and ketamine anesthesia. AB - Extracellular single-unit recording techniques were used to examine the rat globus pallidus (GP). In both locally anesthetized, paralyzed rats and ketamine anesthetized rats, we observed two distinct biphasic extracellular waveforms, which we have labeled Type I (negative/positive waveform) and Type II (positive/negative waveform). No significant differences were observed in the firing pattern or number of cells per track between these cell types, although the Type II neurons had a faster mean firing rate in the locally anesthetized animals. A portion of both cell types could be antidromically activated from the subthalamic nucleus, although Type II neurons had significantly slower conduction velocities. The most striking pharmacological difference between the two cell types was that Type I GP neurons were inhibited by systemic administration of the dopamine agonist apomorphine; previous studies have repeatedly shown that Type II GP cells are excited by this treatment. Pretreatment with a subthreshold dose of apomorphine reduced the responsiveness of Type I cells to a subsequent high dose of apomorphine, as has been shown for Type II cells. However, pretreatment with the NMDA antagonist dizocilpine (MK801) produced a significant change in the pattern of response to apomorphine for Type II GP neurons only. Relative to observations in locally anesthetized, paralyzed rats, ketamine anesthesia reduced the firing rate of both cell types, but did not significantly alter their direction of response to apomorphine. Thus, this study has confirmed the existence of two GP cell types with distinct extracellular waveforms and different responses to dopamine receptor stimulation. These data may necessitate a reevaluation of general theoretical models of basal ganglia function in order to account for these opposite effects of dopamine receptor stimulation on pallidal output. PMID- 7482292 TI - D3 and D2 dopamine receptors: visualization of cellular expression patterns in motor and limbic structures. AB - The distribution of the D3 and D2 dopamine receptor subtypes in forebrain regions of the basal ganglia and mesocorticolimbic system was determined. This was assessed through combined fluorescent visualization of subtype selective anti peptide antibodies for these cloned receptors and detection of their ligand recognition sites using the D2 subfamily antagonist,N-(p-aminophenethyl) spiperone (NAPS fluoroprobe). The double-labeling technique enabled direct comparison of the cloned receptor proteins and NAPS fluoroprobe binding in vitro. The application of these two methods together produced results comparable to single-labeling paradigms. Functional D3 receptors, defined as the coincident fluorescence of the D3 receptor antisera and fluoroprobe binding, were detected in the core region of the nucleus accumbens and exhibited a laminated expression pattern in the frontal cortex. D3 receptor protein was expressed robustly in neurons of the dorsolateral striatum, but showed an intense neuropil reaction in the globus pallidus. Functional D2 receptors, defined as the coincident fluorescence of the D2 receptor antisera and fluoroprobe binding, were detected in the frontal cortex and the medial shell of the nucleus accumbens. Thus, heterogeneities occurred in the cellular expression of functional D3 and D2 receptors in forebrain dopaminoceptive areas. D3 appears more related to basal ganglia and structures involved with motoric behavior, while D2 was associated with regions associated with cognitive/affective functions. PMID- 7482295 TI - Neurotensin increases endogenous glutamate release in the neostriatum of the awake rat. PMID- 7482294 TI - Comparison of the action of the stereoisomers of the psychostimulant 4 methylaminorex (4-MAX) on midbrain dopamine cells in the rat: an extracellular single unit study. AB - In this study, we examined and characterized the action of the stereoisomers of 2 amino-4-methyl-delta 2-5-phenyl-oxazoline (4-methylaminorex, 4-MAX) on spontaneously active dopamine (DA) neurons in the substantia nigra pars compacta (SNC or A9) and ventral tegmental area (VTA or A10) in anesthetized male rats. This was accomplished using the technique of extracellular single unit recording. The intravenous (i.v.) administration of the stereoisomers of 4-MAX (0.1-6.4 mg/kg) produced a dose-dependent suppression of the basal firing rate of A10 DA cells with the following rank order of potency: trans 4S,5S > cis 4R,5S approximately cis 4S,5R >> trans 4S,5S 4-MAX. The rank order of potency of the isomers of 4-MAX to suppress the firing of A9 DA cells was trans 4S,5S = cis 4R,5S = cis 4S,5R >> trans 4R,5R. The trans 4S,5S isomer was 5-fold more potent in suppressing DA cell firing in the A10 compared to the A9 area. The suppressant action of the isomers on A9 and A10 DA cells was reversed by the i.v. administration of haloperidol and the D2/D3 receptor antagonists (-)-sulpiride and (-)-eticlopride but not by the D1 receptor antagonists SCH 23390 and SCH 39166. In addition, the suppressant action of the trans 4S,5S isomer on A10 DA cells was not antagonized or reversed by the i.v. administration of the receptor antagonists granisetron (5-HT3), ritanserin (5-HT2A,C), idazoxan (alpha 2), phentolamine (peripheral alpha 1), (+/-)-pindolol (5-HT1A,B beta) or prazosin (alpha 1). The pretreatment of animals with either alpha-methyl-p-tyrosine (AMPT) or reserpine, but not p-chlorophenylalanine (PCPA), (+/-)-fluoxetine or tomoxetine, significantly attenuated the suppression of A10 DA cell firing produced by trans 4S,5S 4-MAX. Overall, our results suggest that the suppressant action of 4-MAX on midbrain DA cell firing may be mediated by the release of DA, which subsequently interacts with D2/D3 receptors. PMID- 7482296 TI - [Megakaryocytic dysplasia in acute nonlymphoblastic leukemias]. AB - Megakaryocytic dysplasia, platelet and megakaryocytic counts were measured in 87 ANLL patients. High megakaryocytic levels were registered in 16.1%, normal in 17.2%, low or negligible in the rest of the examinees. Half of the patients had dysplasia. Thrombocytopenia present in 93% of cases attested to ineffective proliferation of megakaryocytes. Increased number of megakaryocytes with signs of dysplasia occurred more commonly in M0- and M4- variants of ANLL universally in anomalies of chromosome 3 long arm with involvement of q21 and/or q26 segments and occasionally in combination with other chromosome disorders. Megakaryocytosis developed less frequently in patients with ANLL variant M2 with t (8;21) and did not correlate with granulocyte and erythroid cell dysplasia. Pronounced megakaryocytosis in combination with thrombocytosis emerged in 4 patients: 2 of them had typical anomalies of chromosome 3--inv3(q21q26) and dup (3q21q26), 1 had monosomy 7 and 1 normal karyotype. Potential mechanisms responsible for dysplasia of megakaryocytes in ANLL are considered. PMID- 7482298 TI - [A "primitive" variant of the blast crisis in chronic myeloleukemia]. AB - While immunotyping blast cells from 45 patients with CML blast crisis, we detected 5 cases with immunologically primitive blast cells. The immunological phenotype of these cells corresponded to that of primitive stem cells which are characterized by expression of CD34 and HLA-DR antigens in the absence of other immunological markers. We suggest that blast cells from these patients may undergo differentiation similar to that of primitive stem cells that implies the existence of a new immunological variant of CML blast crisis, a primitive variant. Morphologically, blast cells in 3 cases could be classified as myeloid, in 2 cases precise identification was impossible. Cytochemically, this type of cells can be defined as mixed. The patients with CD34+ phenotype do not differ clinically or hematologically from those with CML blast crisis. Blast cells with membrane marker CD34 are likely to arise in any CML phase either as a component of overall leukemic population or predominant, single subclone. PMID- 7482297 TI - [The clinical significance of the expression of the multiple-drug resistance protein P-glycoprotein in chronic myeloleukemia]. AB - Cell resistance to pharmaceutical agents arises among other causes because of multiple drug resistance induced by P-glycoprotein (P-gp). The analysis of expression of P-gp and differentiation antigens of hemopoietic cells has been made on myeloid cells from 14 patients in CML chronic phase and 25 with CML acceleration and in blast crisis. Surface antigen expression was evaluated at flow cytofluorimetry (FACScan unit). Fluorescent dye rodomin (Rh123) helped examine P-gp functional activity. A close relationship is shown between P-gp expression and CD34 (r = 0.69. p = 0.0004), this giving evidence of these antigens expression on the same cells. In chronic phase P-gp is expressed on a few cells in some patients, its activity being low or absent. The appearance of UIC-2+ cells was unrelated to previous chemotherapy and brought no resistance to treatment. In terminal stage P-gp is expressed in 50% of cases. Functional tests identified the active protein in blast populations with a large number of UIC-2+ cells and in some patients with a small number of cells expressing P-gp. Therefore, comprehensive clinical investigations are needed of multiple drug resistance, though in half of the resistant patients in AML blast crisis P-gp+ cells were not identified suggesting the existence of other mechanisms responsible for resistance to treatment. PMID- 7482299 TI - [The diagnosis of a hemorrhagic syndrome in chronic myeloleukemia]. AB - A study was performed of 40 patients with chronic myeloid leukemia. Clinicohematological, radionuclide (51CR), electron-microscopic methods and examinations of plasma and platelet components of hemostasis were applied to elucidate stage-specific relationships between functional-morphological changes in platelets and early manifestations of hemorrhagic syndrome (HS). The latter was directly related to platelets functional activity and their ultrastructure. With HS progression, platelet functions decline, their degeneration becomes more evident in line with growing blood loss from gastrointestinal tract. PMID- 7482300 TI - [The principles of the differential diagnosis of mature-cell lymphatic tumors]. PMID- 7482301 TI - [The clinical significance of the immunological phenotype in non-Hodgkin's lymphomas]. AB - To study immunological phenotype of tumor cells in the blood and tumor tissue of 68 patients with non-Hodgkin's lymphomas, the authors have used flow cytometry of lymphocytes and enzyme immunoassay in cryostat sections of lymph nodes with application of monoclonal antibodies to differentiating antigens of human lymphocytes. Histological types of lymphoma are shown to differ immunophenotypically. Differential antigens of prognostic and diagnostic value have been determined. PMID- 7482302 TI - [Changes in the intraerythrocytic proteolysis of human hemoglobin in lymphogranulomatosis]. AB - The data obtained on red cell hemolysates from 16 patients with Hodgkin's disease varying in age, sex, the disease stage and histology, time since the diagnosis served the basis for comparative evaluation of endogenic intraerythrocytic hemoglobin proteolysis and relevant products. The results show that Hodgkin's disease brings about a 10-15-fold increase in the quantity of accumulated hemoglobin alpha-chain fragments. PMID- 7482303 TI - [The significance of lactate dehydrogenase and beta 2-microglobulin levels for the assessment of the prognosis and choice of therapy in multiple myeloma]. AB - Serum levels of lactate dehydrogenase (LDG) and beta 2-microglobulin (beta 2-MG) were measured in 164 and 128 patients with multiple myeloma (MM), respectively. High levels of LDG were recordable in 15.4% of patients at diagnosis and 36.8% of terminal stage patients. The frequency of extraosseous foci in untreated patients with high LDG activity made up 36.8%, survival median 19 months. In normal LDG activity the above values were 6.8% and more than 36 months, respectively. The highest LDG level occurred in patients with terminal plasmic cell leukemia. MM with IgD secretion was characterized by a a more frequent rise in LDG concentrations. Normal LDG amounts in active MM were seen in 58 (54.2%) out of 107 patients. beta 2-MG levels exceeded 6 mg/l in 75 of 128 patients with normal creatinine. These patients had a short survival median 24 months. Those patients who had beta 2-MG levels under 6 mg/l have not reached survival median for 36 months of follow-up. The authors hold that beta 2-MG concentrations are of prognostic value in all myeloma secretions and in nonsecretory myeloma as well though their indications are not absolute as 14% had low beta 2-MG levels in high MM activity. Comparative results are presented for 40 high-risk MM patients. Group 1 (20 patients) have received standard chemotherapy. Group 2 (20 patients) have undergone intensive polychemotherapy. Survival median made up 12 and 26 months for group 1 and 2, respectively. PMID- 7482304 TI - [The prolymphocytic-lymphocytic leukemization of T-cell lymphosarcomas]. AB - The paper presents a detailed clinical, hematological, morphological, ultrastructural and immunological characterisation of T-cell lymphosarcoma with prolymphocytic-lymphocytic leukemic transformation (PLLT). In PLLT the proportion of T-cell immunological subvariant of lymphosarcoma came to 15% being detected only in 8 out of 52 examinees. The patients (6 males and 2 females) varied in age from 24 to 76 years (median 49 years) and had the following histological forms of primary tumor tissue: lymphoblastic lymphosarcoma (n = 3), pleiomorphic small cell lymphosarcoma (n = 1), large-cell anaplastic lymphosarcoma (n = 1), prolymphocytic lymphosarcoma. Immunological characteristics of these 8 cases were heterogeneous: in lymphoblastic variant there was immature T-immunological phenotype. In pleomorphic small-cell lymphosarcoma there were also signs of T cell activation. In large-cell anaplastic lymphosarcoma an immunological phenotype of lymphoid cells from the primary tumor tissue and bone marrow differed in more advanced immunological differentiation of bone marrow tumor cells. In prolymphocytic variant tumor cells had features of T-helpers or T suppressors. Most of the patients received polychemotherapy according to the schemes for high-grade lymphosarcomas despite PLLT though the latter is not a universal indicator of late tumor progression, more aggressive course of the disease and poor prognosis. PMID- 7482305 TI - [Errors in the diagnosis and therapy of chronic lympholeukemias]. AB - The analysis of 67 cases of benign, progressive, tumor CLL, lymph node lymphocytoma, CLL sarcoma transformation showed that the cases were misdiagnosed in 35.8% cases. The most common mistake (25.4%) was aggressive chemotherapy based on histological diagnosis of prolymphocytic, prolymphocytic-lymphoblastic lymphosarcoma without consideration of cytological and clinical evidence, tumor phenotype. The authors think valid to use the following criteria in diagnosis of lymphoproliferative diseases: histological findings, clinical manifestations and blood picture, tumor cell cytology, primary location and predominant dissemination of the tumor, immunophenotype, characteristic chromosomal disorders, response to treatment. PMID- 7482306 TI - [Thrombophilia characterized by resistance to anticoagulants with indirect action]. AB - The paper reports 2 cases of resistance to indirect-action anticoagulants used for prevention of thrombosis. One female patient combined resistance to fenilin with that to aspirin and trental, platelet hyperaggregation returned to normal due to ibustrin. The change of fenilin for pelentan did not influence the situation. In another case the resistance was to fenilin, but hypocoagulation was achieved with neodicumarin in standard doses. Platelet hyperaggregation was corrected by routine doses of aspirin. Causes of resistance to anticoagulants acting indirectly are discussed. PMID- 7482307 TI - [Acute respiratory failure in hemoblastoses: the types of diffuse lung lesions]. PMID- 7482308 TI - [The sonographic indices of the spleen in different diseases of the blood system]. PMID- 7482309 TI - [A trial of the use of sandostatin in patients with chronic pancreatitis]. AB - Out of 15 patients with chronic pancreatitis (CP) treated with sandostatin, 8 patients demonstrated a complete and 6 partial response. One patient did not respond. Pain relief occurred in all of them. Side effects were registered in 3 patients (doughy stools 4 times a day throughout treatment). Normal blood levels of pancreatic enzymes, insulin secretion, parameters of blood inhibitory system did not change much because of sandostatin administration, whereas hypercoagulation got diminished. Rat experiments have revealed a trend to trypsin lowering in tissues of unaffected pancreas and more intensive inhibition of active trypsin by tissue inhibitors. PMID- 7482310 TI - [Sandostatin in the treatment of diarrhea of different origins]. AB - Antidiarrheal action of sandostatin (octreotide) was studied in dumping syndrome, diabetic enteropathy, short-bowel syndrome, Crohn's disease and ulcerative colitis. Sandostatin proved effective in diarrhea of secretory, motor and osmotic origin. Dumping syndrome, diabetic enteropathy, Crohn's disease and short-bowel syndrome responded completely. In ulcerative colitis the effect was paradoxical: intensive diarrhea discontinued, but moderate diarrhea became more intensive. Sandostatin indications in ulcerative colitis should receive further investigations. PMID- 7482311 TI - [The potentials for using sandostatin in gastroenterology (a review)]. PMID- 7482312 TI - [The outcome and course of IHD in patients with normal and altered coronary arteries under 10-15 years of observation]. AB - The course and prognosis of ischemic heart disease (IHD) were studied in 175 anginal IHD patients regularly examined coronarographically for 10-15 years. The clinical, bicycle ergometry and coronarography findings have given evidence for design of prognostic risk criteria. The first 3 years of the follow-up have shown cases of lethal and transmural myocardial infarction to occur only in high-risk group. Low-risk patients developed transmural myocardial infarction on follow-up year 7, while lethal outcome occurred 14 years after the start of the observation. In patients with normal or insignificantly changed coronary arteries the above events were not observed. PMID- 7482313 TI - [Corticosteroid therapy and aseptic bone necrosis in patients with rheumatoid arthritis]. AB - Two groups of RA patients were compared: 102 patients with osteonecrosis of the head of the femur (group 1) and 50 patients free of it (group 2). Group 1 patients had bilateral lesion of the hip joints (28% of cases), unilateral osteonecrosis of knee joint in 14.5% and bilateral in 2.9% of the patients. Unilateral humeral osteonecrosis occurred in 17.6%, bilateral in 4.9% of patients. Extra-articular manifestations of RA (lymphadenopathy, renal lesions and Cushing's syndrome) were diagnosed more frequently in group 1. Progression of osteonecrosis is related to a mean maximal day dose of prednisolone in the first month of therapy. PMID- 7482314 TI - [The history of the study of the pathogenesis and the search for agents for the treatment of cholera (based on data from the first 3 pandemics of 1817-1862)]. PMID- 7482316 TI - [High-dose therapy with autologous bone marrow transplantation in patients with resistant forms of lymphogranulomatosis]. PMID- 7482315 TI - [The results of 2 and 1/2 years of work of the Russian Multicenter Trial on the Treatment of Acute Myeloid Leukemias in Adults]. PMID- 7482317 TI - [Hematological masks in the internal disease clinic]. PMID- 7482318 TI - [Complications in hemotherapy]. PMID- 7482319 TI - [The therapy of mesangiocapillary glomerulonephritis]. AB - Follow-up of 7 patients with nephritically active mesangiocapillary chronic glomerulonephritis which received for 22 months combined therapy with prednisone (60 mg), chlorambucil (0.2 mg/kg), curantyl (400 mg), heparin or fenilin has found a significant lowering of proteinuria, hypercreatinemia, a rise in osmotic concentration. A complete remission was achieved in 2 patients, partial in 3 patients, a progression occurred after 8 years in one. PMID- 7482320 TI - [The treatment of nephrotic-type chronic glomerulonephritis with Sandimmun (cyclosporin A)]. AB - Sandimmun (ciclosporin A) treatment of chronic glomerulonephritis with nephrotic syndrome in a dose 2.5-5 mg/kg for 6-10 months relieved nephrotic syndrome in 2 and resulted in clinical improvement in other 2 of 4 patients treated. In 3 cases the patients failed previous therapy with high-dose corticosteroids and cytostatics. The drug reduced proteinuria, elevated serum levels of albumin and total protein, had low nephrotoxicity. PMID- 7482321 TI - [Antihyperlipidemic actions in the treatment of nephropathies]. AB - In view of weak implication of immunoinflammatory mechanisms in pathogenesis of noninflammatory nephropathy one may expect the disease course improvement and more slow progression of renal dysfunction. The mechanism of this action can be realized through hypolipidemic effect. In recent research there are data on a direct lovastatin action on the mechanism underlying nephritis progression by means of inhibiting proliferation of mesangial cells. PMID- 7482322 TI - [Primary chronic pyelonephritis and kidney anomalies. The characteristics of their diagnosis, course and outcomes (the author's own observations)]. PMID- 7482323 TI - [The effect of the early treatment of patients with hemorrhagic fever with renal syndrome on the further course of the disease]. AB - 173 case records have been analyzed to specify the role of early therapy (antipyretics, antibiotics, sulphanilamide drugs) on further development of hemorrhagic fever with renal syndrome. Early treatment of the above patients was found uneffective and not safe as to possible complications, for instance antipyretics occasionally brought hypotonia and shock. PMID- 7482324 TI - [The factors affecting the rehabilitation of patients on chronic hemodialysis treatment]. PMID- 7482325 TI - [Anomalous chordae tendineae]. AB - ECG of 322 patients with various cardiovascular diseases allowed the conclusion on the occurrence of anomalous chordae of the heart in 21.7% of cases. The chordae had no effect on the disease diagnosis, running, hemodynamics, left ventricular myocardial mass. PMID- 7482326 TI - [The physicochemical properties of molsidomine preparations and their efficacy in treating IHD and circulatory failure]. AB - Molsidomine drugs corvaton and sidnopharm were assessed clinically, physicochemically and experimentally. Corvaton tablets contained more active substance but less cleavage products than sidnopharm ones. In experiment with acute transitory coronary insufficiency corvaton was active both during myocardial ischemia and in reperfusion. Corvaton showed antianginal effect in 70.3%, sidnopharm in 58.6% of cases. Both drugs produced a positive action on coronary and myocardial reserves in coronary patients with angina class I-III but did not affect hemodynamics. Oral sidnopharm induced side effects 4 times more frequently than corvaton. Corvaton tablets (single dose) activity lasted more than sidnopharm effect by 2.2 hours. Oral corvaton remained active 5.3 hours longer than the same intravenous dose. PMID- 7482327 TI - [The lipid peroxidation indices of hypertension patients undergoing combined therapy with anaprilin, korinfar and hypothiazide]. AB - Lipid peroxidation (LPO) products and activity of antioxidant defense (AOD) were evaluated in the whole blood and red cells of 78 patients with hypertension stage II given anapriline, corinfar and hypotiazid. Combined therapy with anapriline and hypotiazid (80-120 mg/day and 50 mg/day, respectively) diminishes concentrations of diene conjugates and malonic dialdehyde (MDA) in blood though LPO parameters after the treatment exceeded those of healthy subjects. Native antioxidants content and activity of antioxidant enzymes glutathione peroxidase and superoxide dismutase increased. Combination of corinfar (30-60 mg/day) with anapriline (80-120 mg/day) inhibits LPO and antioxidant defense activity to optimal leading to normalization of some LPO values and activity of antioxidant enzymes after the treatment. This combination was also effective as related to lowering of high blood pressure. PMID- 7482328 TI - [Diabetic and gouty nephropathies (certain aspects)]. PMID- 7482329 TI - [The effect of beta-adrenoblockaders on the lipid composition of the blood]. PMID- 7482330 TI - [Secondary prevention and the rehabilitative therapy of patients with different forms of coronary pathology. The individual approach and criteria for determining efficacy]. PMID- 7482331 TI - [The effect of clinical and hemodynamic factors on the dynamic development over the course of 1 year of observation of heart failure in rheumatism patients]. AB - Factors contributing to congestive heart failure were specified in a 1-year follow-up of 108 patients with rheumatic heart disease. Relevant clinical factors comprised the kind of valvular defect, type of hemodynamic disorder and the degree of circulatory insufficiency. Among instrumental indices of importance were cardiac preload and myocardial contractility. Positive changes occurred in the use of peripheral vasodilators affecting primarily the venous vascular bed. The prognosis got deteriorated due to digitalis intoxication. The authors emphasize the necessity of more careful follow-up of rheumatic heart disease patients at high risk of progressive heart decompensation. PMID- 7482332 TI - [The hemodynamic status and right ventricular diastolic function of bronchial asthma patients]. AB - Doppler echocardiographic study of ventricular diastolic function in 90 BA patients gave evidence for diastolic left and right ventricular dysfunction in patients with moderately and gravely running BA. The above disorders were noted by changes in the speed of transmitral and transtricuspid flows: early filling rate (E) slowing, atrial systole filling rate (A) acceleration, E/A decrease. The degree of right ventricular diastolic dysfunction depends on right ventricular hypertrophy and total pulmonary resistance. Left ventricular diastolic function is related to right ventricular after-load. PMID- 7482333 TI - [Bronchial nonspecific reactivity in patients with bronchial asthma and in the preasthmatic state and its alteration under the influence of acupuncture]. AB - The development of nonspecific bronchial hypersensitivity and hyperreactivity in bronchial asthma and effectiveness of its correction with acupuncture were investigated in 152 patients with asthma and preasthma. 94 of them were subjected to acupuncture. The authors employed a complex of diagnostic methods with determination of 241 parameters processed later with the use of systemic modelling. Bronchial hypersensitivity as indicated by the response to acetylcholine was related to impaired coordination of bronchomotor tone regulating systems: parasympathetic part of the autonomic nervous system, eosinophilic and monocytic bronchial inflammation, glucocorticoid homeostatic alterations. The corrective role of acupuncture consists in reduction of nonspecific bronchial hyperreactivity, normalization of blood acetylcholine, resensitization of cell beta-adrenergic receptors, elevation of mean concentrations of 11-OCS and T-lymphocytes. PMID- 7482335 TI - [The mechanisms of the development of status asthmaticus]. AB - The author provides evidence confirming the fact that disturbance of mucociliary clearance (mucobronchostasis) because of chronic bronchitis and repetitive contacts with allergens underlies asthmatic condition. Hypophysis on leading role of B2-adrenoreceptor block in pathogenesis of asthmatic condition was not confirmed as the block occurs occasionally and depends on the disease form and severity. PMID- 7482334 TI - [Disorders in mineral metabolism at different stages of the development of bronchial asthma]. AB - Magnesium and calcium concentrations were measured by nuclear-absorption spectrometry (AAC-508, Hitachi, Japan) and selective ionometry (Microlyte, Kone, Finland) in the serum, red cells, lymphocytes, 24-h urine and exhaled air humor condensate in 23, 31 and 23 sibs (parents and their children) with preasthma, atopic bronchial asthma, free of these diseases, respectively. All the patients had magnesium deficiency and calcium hyperconcentrations in the blood cells, low urine excretion of magnesium. Parathormone and calcitonin serum concentrations underwent no changes. It is suggested that in contrast to changes of endobronchial homeostasis of bivalent cations, those of electrolyte cell composition are primary and appear prior to clinical manifestations of bronchial asthma. PMID- 7482336 TI - [The immunogenetic heterogeneity of chronic glomerulonephritis]. AB - Clinico-immunogenetic investigations show that there exists highly significant positive and negative associations of HLA-antigen with chronic glomerulonephritis (CGN). The detected HLA markers of CGN predisposition may appear valuable for grouping subjects at high risk to develop CGN, in prognostication of individual features of CGN running, differential diagnosis and perfection of early therapy. PMID- 7482339 TI - [The HLA-associated immunological mechanisms in rheumatic involvement of the myocardium]. AB - The examination of 95 Uzbeks with rheumatic myocardium and 60 healthy subjects of the same nationality has found accumulation in the patients' phenotypes of antigens HLA-B17, HLA-B21 and HLA-Cw4. A high PHA-induced response of T lymphocytes occurred in HLA-Cw4- and especially in HLA-B21-positive patients. The findings suggest participation of HLA-associated genes in the mechanisms of rheumatic process. These are thought responsible for rheumatism-specific cell immune reactions. PMID- 7482338 TI - [Antibodies to skeletal muscle myofibrillar proteins in the diagnosis of rheumatoid arthritis]. AB - One of the mechanisms of muscular involvement in RA patients was elucidated by identification of antibodies to skeletal muscle components (actin and tropomyosin). Clinical symptoms of skeletal muscles involvement in RA proved to be associated with elevated count of tropomyosin antibodies against an insignificant informative value of actin. Enzyme immunoassay with tropomyosin has established a direct correlation between the number of the antibodies and RA activity. PMID- 7482337 TI - [The treatment results in patients with aplastic anemia using different doses of antilymphocyte globulin]. AB - The authors present 5-year survival of 64 patients with aplastic anemia (AA) divided into 2 groups. Group 1 received antilymphocytic globulin (ALG) in low doses (1-5 mg/kg), group 2 in high doses (10-30 mg/kg). Survival of patients with severe AA in group 1 made up 28.5%, with mild AA 90.1%. Overall survival in group 1 was 64.4%, in group 2 53.8%. Overall toxicity came to 48.6%. By resultant complications high and low ALG doses did not differ. The authors propose to use low doses of ALG in mild disease whereas in severe condition high ALG doses are thought adequate. PMID- 7482340 TI - [The combined intensive therapy of rheumatoid arthritis with systemic manifestations]. AB - The efficacy of pulse therapy in combination with hemosorption or plasmapheresis, pulse therapy without extracorporeal treatment and methotrexate has been compared for 40 patients with rheumatoid arthritis (RA) with extra-articular manifestations. All kinds of intensive treatment were effective. Extracorporeal methods and pulse therapy relieved extra-articular symptoms. Isolated pulse therapy alleviated articular syndrome. The best results were obtained in double filtration of plasma in combination with pulse therapy. Such approach ensured improvement both articular and extra-articular inflammation. Combined intensive therapy proved an effective modality in RA able to produce responses in severe disease. PMID- 7482341 TI - [Permixon in the treatment of benign prostatic hyperplasia]. PMID- 7482342 TI - [Cases of Reiter's disease in twin brothers]. PMID- 7482343 TI - [The effect of angiotensin-converting enzyme inhibitors on left ventricular hypertrophy in patients with arterial hypertension]. PMID- 7482347 TI - [Dynamic bone marrow take and lymphohematopoietic chimerism following allogeneic bone marrow transplantation]. PMID- 7482344 TI - [The prostanoid system in patients with disordered heart rhythm]. PMID- 7482346 TI - [The role of hypoxia in the development of bronchial asthma]. PMID- 7482345 TI - [The erythrocyte pro-oxidant and antioxidant systems of patients with chronic kidney failure]. AB - Lipid peroxidation as shown by malonic dialdehyde (MDA) levels and enzymic antioxidant defense systems were evaluated in red cells from patients with renal affections free of chronic renal failure (group 1), in conservative curable stage of chronic renal failure (group 2a), in terminal stage nondialysis patients (group 2b) and in healthy donors. MDA was higher in patients, in group 2b in particular. MDA levels correlated with concentrations of endogenic creatinine in the serum. Catalase and glutathione peroxidase were at control levels. SOD was not changed in group 1 but appeared reduced in other groups. Its activity was not related to serum creatinine. An inverse relationship existed between MDA content and SOD activity in red cells. It is believed that progression of chronic renal insufficiency leads to activation of lipid peroxidation and deterioration of antioxidant defense in red cells contributing to more active red cell destruction causing anemia in uremia. PMID- 7482348 TI - [The current treatment of asthma and chronic obstructive pulmonary diseases (an international consensus on diagnosis and treatment)]. PMID- 7482349 TI - Increased risk of craniosynostosis with maternal cigarette smoking during pregnancy. PMID- 7482350 TI - Antisense targeting of engrailed-1 causes abnormal axis formation in mouse embryos. AB - Antisense oligonucleotide targeting of engrailed-1 (En-1) in early somite mouse embryos resulted in reduced En protein levels and produced abnormalities of the brain, face, and heart and shortening of the embryonic axis (caudal dysgenesis). Defects of the brain and limbs were consistent with abnormalities observed in null mutant mice described by other investigators. Abnormalities of the face and heart may be related to alterations in neural crest cells. Caudal dysgenesis suggested a role for En-1 in axis formation and this hypothesis was supported by results showing that En-1 protein and mRNA are present in the primitive streak. Thus, in addition to participating in the signaling pathway for brain and limb development, En-1 appears to play a role in patterning the embryonic axis. PMID- 7482351 TI - Antisense inhibition of engrailed genes in mouse embryos reveals roles for these genes in craniofacial and neural tube development. AB - The roles of engrailed-1 (En-1) and engrailed-2 (En-2) have been investigated during gastrulation and neurulation in mouse embryos. Using antisense oligonucleotides and murine whole embryo culture, early somite embryos were injected with antisense phosphorothioated oligonucleotides directed against En-1 or En-2 transcripts and then grown in vitro for 48 hr. Inhibition of En-1 by antisense targeting during this period resulted in embryos with craniofacial abnormalities, specifically loss of mid- and hindbrain tissue and hypoplasia in associated neural crest derived areas such as the face and first and second pharyngeal arches. In addition, En-1 appeared to be essential in early patterning of the neural tube. Embryos removed from culture as early as 8 hr following injection exhibited undulations in the presumptive spinal cord. Histological analysis of the affected neural tubes at 48 hr showed disrupted cytoarchitecture and in some cases, apparent dorsal-ventral duplication of the neural tube and underlying notochord. Using S100 beta as a notochord marker, embryos removed from culture at 8 or 12 hr following injection exhibited loss of S100 beta expression in the notochord region subjacent to affected neural tube segments, suggesting that S100 beta, or other notochordal genes, may be downstream components of the En-1 regulatory cascade. Furthermore, antisense inhibition of En-1 induced caudal dysgenesis, suggesting disruption in primitive streak function. Antisense targeting of En-2 expression of early somite stages produced few alterations in development, although approximately one third of the embryos exhibited a series of lateral folds in the spinal cord at the level of the forelimb-bud.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7482352 TI - Elevated midtrimester serum methylmalonic acid levels as a risk factor for neural tube defects. AB - The role of folic acid in the primary prevention of neural tube defects (NTDs) is well established. However, questions related to the protective mechanism remain unanswered. To help answer these questions, we designed a case-control study to assess the role of folate- and cobalamin-related metabolites in the pathogenesis of NTDs. Concentrations of folate, cobalamin, and 14 other related metabolites were measured by gas chromatography/mass spectrometry in midtrimester serum specimens from 32 women with an NTD-affected pregnancy and from 132 control women, and in serum specimens from 46 nonpregnant women who had a history of NTD affected pregnancy and from 43 nonpregnant control women. Log-transformed means of metabolites were compared between case and control women for both the midtrimester and nonpregnant groups. In the pregnant group, serum methylmalonic acid (MMA) concentrations were higher among case women than among control women (130 vs 105 nM). There was a strong dose-response relationship between midtrimester serum MMA level and the risk for an NTD-affected pregnancy, with the relative risk increasing 13-fold for women with MMA levels > 90th percentile. In the nonpregnant group, there was no difference in serum MMA levels between case and control women (140 vs 140 nM). Thus, the serum MMA levels of women in the midtrimester of pregnancies unaffected by NTDs were significantly lower than the levels of nonpregnant women, whereas the levels of women whose pregnancies were affected by NTDs were similar to those of nonpregnant women. The finding of elevated MMA serum concentrations among women in the midtrimester of NTD-affected pregnancies suggests that cobalamin may be involved in the etiology of NTDs. The possible role of cobalamin in relation to the protective effect of folic acid needs further evaluation. PMID- 7482353 TI - Effects of prenatal alcohol exposure on the postnatal morphology of the rat oculomotor nucleus. AB - Morphological development of the rat oculomotor nucleus was investigated on postnatal day 15 following a prenatal ethanol exposure. Analysis of toluidine blue stained plastic sections showed that the prenatal alcohol exposure caused a decrease in the density of neurons and an increase in the density of astrocytes in the center of the nucleus. There was an alcohol-induced reduction in the overall size of the cross-sectional region of the oculomotor nucleus, but no effect on the number of neurons per unit area of that total oculomotor region, indicating a delay or alteration of the migration of neurons to their normal clustered position in the center of the nucleus. The areas of the neuronal cell nucleus and nucleolus were not affected by the alcohol exposure. Analysis of Golgi-Cox-impregnated multipolar neurons showed that the alcohol exposure caused a reduction in area of the cell soma; a reduction in the number of dendritic branches; and a reduction in the complexity of the dendritic arbor relative to distance from the soma, based on concentric ring analysis. The results of this study demonstrate that gestational alcohol exposure can retard the maturation of the oculomotor nucleus. PMID- 7482354 TI - Anti-interferon immunoglobulins can improve the trisomy 16 mouse phenotype. AB - Mouse trisomy 16 is a well-studied model for human chromosome 21 trisomy (Down's syndrome). The late stage trisomy 16 mouse fetus exhibits significant growth retardation, inappropriately opened eyes, and convex rather than concave back curvature. The interferons (alpha, beta, and gamma) have potent growth retarding activity, and sensitivity to these cytokines is controlled by genes that map to mouse chromosome 16 and human chromosome 21. In experiments designed to determine if the interferons induce or aggravate the trisomy phenotype, mice pregnant with trisomy 16 fetuses were injected with a combination of anti-alpha, -beta, and gamma interferon IgG. This maternal anti-interferon treatment was found to provide measurable benefit to the development and growth of the trisomic fetuses with significant return-toward-normal values observed for overall fetal growth, eye opening, and back curvature. PMID- 7482355 TI - Detection of genotype-environment interaction in case-control studies of birth defects: how big a sample size? AB - Detecting interactions between risk factors in case-control studies of birth defects and other conditions usually requires increasing the sample size beyond that needed to detect marginal effects. A special case of such interaction is genotype-environment interaction in which the effects of an exposure on disease risk are modified by genetic susceptibility. When case-control studies are designed to detect marginal effects of an exposure (i.e., in the whole population), under many plausible interaction schemes, no additional case and control subjects are needed to detect genotype-environment interaction. On the contrary, inclusion of genotypic information generally can improve the statistical power of the original study. Using the example of oral clefts, maternal cigarette smoking, and genetic variation at the transforming growth factor alpha gene, we illustrate sample size and power issues in designing case control studies when prior information is available on both the marginal effects of the exposure and the genetic factor. PMID- 7482356 TI - Teratogen update: ergot and ergotamine. AB - Ergotamine is a purified ergot alkaloid that has been suspected of being teratogenic. Animal data have shown effects mainly at doses toxic to the mother. Clinical reports in humans have been anecdotal, but in many the malformations are consistent with vascular injury. However, epidemiologic studies have not shown any clear increase in malformations among exposed infants. This may reflect the limited exposure and toxicity when used episodically. Ergot toxicity which can result in severe vasospasm does not spare the fetus or mother and ergotamine should not be considered without risk. Ergotamine should be avoided in pregnancy, and the counselling of the woman with an exposed fetus should focus on possible toxic exposure. PMID- 7482357 TI - Working together. Physicians, midlevel practitioners reach compromise on scope-of practice issues. AB - When the 74th Texas Legislature convened last January, lobbyists for the Texas Medical Association anticipated a bitter fight over attempts by advanced nurse practitioners to gain authority to independently prescribe drugs. After all, physicians and nurses had been fighting for several sessions over scope-of practice issues. Near the end of the 73rd Legislature, in 1993, nurse practitioners (NPs) stalked away from the bargaining table, effectively killing a compromise that some thought was close to approval. PMID- 7482358 TI - Managing Medicaid. Impending change produces many questions, few answers. AB - There is a widely held belief that big trouble often starts with a simple declaration: "Now here's what we'll do." It's a variation of the one about hell and good intentions. During the most recent session of the Texas Legislature, lawmakers decided that what they would do is authorize the conversion of the state's Medicaid program to managed care. Otherwise, Texas faced a projected shortfall of up to $2 billion in state funds over the next biennium for the program that provides health care for about 2 million low-income Texans a month. PMID- 7482359 TI - Legislative soup, rule-making salad. Lawmakers and state agencies serve up 5.01(a) rules. AB - One response to tremendous changes in the health-care industry has been the formation of more and more physician/hospital organizations (PHOs). One kind of PHO has grown rapidly in Texas. Commonly referred to simply as the 5.01(a)--its section number in the Medical Practice Act of Texas that describes how it is formed and what it can do--the number of these nonprofit health-care corporations has more than quadrupled in Texas in the past 5 years, increasing from 36 in 1990 to 160 in 1995. PMID- 7482360 TI - Town versus gown. PMID- 7482361 TI - Treating violence. Future physicians search for answers to a different kind of epidemic. AB - When today's medical students look back at why they decided to become physicians, most say they thought about helping people who were battling diseases, not battling each other. Students going into emergency medicine and psychiatry may have considered the effects of gunshot wounds and terrorist bomb attacks. But the epidemic of violence is also running a more subtle course. Now all physicians, from ophthalmologists to orthopedic surgeons, are on call to watch for evidence of different kinds of abuse and neglect--child, elderly, mental, and physical. And many physicians are not prepared to recognize the signs of domestic violence, direct their patients to helpful resources, or protect themselves against violent attacks. The lack of instruction in these areas prompted the Texas Medical Association Medical Student Section (MSS) to adopt violence as its 1994-1995 social issue. PMID- 7482362 TI - Bike helmet campaign making statewide impact. PMID- 7482363 TI - Don't hold still for miseducation by utilization reviewers. PMID- 7482364 TI - Surgical aspects of epilepsy. 1920. PMID- 7482365 TI - Another view of military medicine. PMID- 7482366 TI - Patient protection rules a good first step. PMID- 7482367 TI - Governor's veto hurts physicians and patients. PMID- 7482368 TI - The fight for our profession. PMID- 7482369 TI - Is Texas ready? Gerontology specialists in health care. PMID- 7482370 TI - Statistical properties of segregating sites. AB - A mutation leading to a segregating site of a sample can be classified by the number of sequences in the sample that inherits the mutant nucleotide; it can also be classified by the frequencies of the two segregating nucleotides at the resulting segregating side. We define the size of a mutation to be the number of sequences in the sample that inherits the mutant nucleotide and the type of mutation (segregating site) to be the smallest value of the frequencies of segregating nucleotides. Each of these two classifications of mutations is analogous to allelic types in a sample of genes. Assuming the neutral Wright Fisher model, we derived in this paper the mean and variance of the frequency of mutations of each size and type, and the covariance between the numbers of mutations of two different sizes and two different types. Potential applications of these results are discussed. PMID- 7482371 TI - Divergence time and population size in the lineage leading to modern humans. AB - We have developed maximum likelihood (ML) methods for comparisons of nucleotide sequences from unlinked genomic regions. In the case of a single species, the ML method primarily estimates the effective population size (Ne) under both constant size and abrupt expansion conditions. In the case of two or three species, the ML method simultaneously estimates the species divergence time and the effective size of ancestral populations. This allows us to trace the evolutionary history of the human population over the past several million years (my). Available sequences at human autosomal loci indicate Ne = 10,000 in the Late Pleistocene, a figure concordant with the results obtained from mitochondrial DNA sequence and allele-frequency data analysis, and there is no indication of population expansion. The ML analysis of two species shows that humans diverged from chimpanzees 4.6 my ago and that the human and chimpanzee clade diverged from the gorilla 7.2 my ago. Furthermore, the effective population size of humans more than 4.6 my ago is nearly 10 times larger than Ne of modern humans. The effective population size in the human lineage does not seem to have remained constant over the past several million years. The ML model for three species predicts slightly different, but consistent results to those obtained by the two-species analysis. PMID- 7482372 TI - [Pharmacotherapy with endogenous substances]. PMID- 7482373 TI - [Intensive insulin therapy--is it worth the effort?]. AB - Diabetes mellitus is associated with a number of well-known, specific macro- and microvascular as well as neuropathic complications. The typical and specific association of microvascular and neuropathic complications with diabetes suggests a causal relationship with hyperglycemia or associated metabolic abnormalities. The results of the Diabetes Control and Complications Trial (DCCT) as well as other recent studies have demonstrated that in patients with insulin-dependent diabetes mellitus (IDDM) the incidence of retinopathy, nephropathy and neuropathy can be reduced by intensive treatment. Strategies of intensified insulin therapy and the clinical importance of improved diabetic control are outlined in view of these studies. PMID- 7482374 TI - [Immunoglobulins--merely expensive or also useful?]. AB - About 15 years ago, treatment with intravenous immune globulin (IVIG) for primary immunodeficiency diseases was introduced. The concept of replacement of deficient circulating antibodies to prevent infections in these patients subsequently proved to be beneficial. Up-to-date IVIG therapy is considered to be the treatment of choice in many primary or secondary immunodeficiency states. The observation of a significant increase in the platelet count in patients with agammaglobulinemia and severe thrombocytopenia after IVIG therapy developed interest in possible modulatory effects of IVIG on the immune system. Although the mode of action of IVIG in autoimmune diseases is not completely understood, therapeutic benefit has been shown in some diseases. It has to be stated that IVIG in autoimmune diseases are rarely first therapeutic choice; however, IVIG might be indicated in patients where conventional and cheaper therapy has failed. This review summarizes the status of IVIG therapy in primary and secondary immunodeficiency states and possible indications in autoimmune diseases. PMID- 7482375 TI - [Interferons as therapy in viral hepatitis: are promises kept?]. AB - Due to their antiviral and immunomodulatory properties, interferons constitute the only efficacious therapy against chronic viral hepatitis. Type-1 interferons are used for therapy with doses ranging between 3 and 10 million units on three days a week. Duration and dose vary with the various forms of viral hepatitis and have not been defined for all instances. In chronic hepatitis B, continued suppression of viral replication can be achieved in 40 to 50% of patients and leads to loss of HBV-DNA and HBe in the serum. 10 to 15% of patients also loose HBs-Ag later on. By carefully selecting patients, even higher success rates can be achieved; however, good selection criteria do not exist for chronic hepatitis C. With this hepatitis, interferons normalize aminotransferases frequently, but sustained elimination of viral infection is still a rare event. Finally, most patients have a relapse of their hepatitis when interferon therapy is terminated. Thus, only 20% of patients with chronic hepatitis C have a sustained benefit from their interferon treatment. Interferon therapy of chronic hepatitis D is even more difficult. Side effects increase with dose and duration of therapy. Fortunately, they only rarely require discontinuation of interferon and are fully reversible, when interferon is stopped. In conclusion, as there exist no therapeutic alternatives, patients with chronic hepatitis B and hepatitis C should be treated with interferon, even though only 20 to 40% of treated patients benefit in the long term. PMID- 7482376 TI - [Fibrinolytic agents--who benefits when?]. AB - Thrombolytic therapy mimics and enhances physiological fibrinolysis. The following substances are presently available for clinical use: the nonphysiological thrombolytics streptokinase, the APSAC (acylated plasminogen streptokinase activator complex), the physiological plasminogen activators urokinase and tissue plasminogen activator (t-PA). Whereas the first three systematically activate the fibrinolytic system, t-PA possesses relative fibrin selectivity. The fibrin-selective active pro-urokinase has not yet been officially approved for the treatment of thromboembolic diseases, but it is being clinically tested. Fibrinolytic therapy has an established place in the management of acute myocardial infarction and of massive pulmonary embolism. When an acute deep venous thrombosis is diagnosed with a proximal extension into the popliteal vein, thrombolytic therapy is clearly superior to heparin. The lysis has proven to be an effective form of treatment of peripheral occlusive arterial disease. Local thrombolytic therapy is an option for acute and chronic femoro popliteal occlusions involving the trifurcation into the calf arteries and for embolic occlusions of the same segment in patients with contraindications to surgical therapy. First study results of thrombolytic therapy of stroke are promising. PMID- 7482377 TI - [Blood thinning in heart patients]. AB - Virtually all modalities of anticoagulation play a role in cardiology. For primary prevention, aspirin showed no effect in a British study, but reduced the incidence of myocardial infarction by 44% per year in an American trial concerning men and by 32% concerning women. In patients with pre-existing coronary artery disease, the effect was even more marked. For unstable angina, several regimens were tested. Aspirin and heparin were the drugs most frequently used, either as single therapy or combined. They both proved efficacious, and so did ticlopidine. Promising results were also obtained with hirudin, a specific thrombin antagonist. For the treatment of acute infarction, the roles of aspirin and vitamin-K antagonists are soundly established. Aspirin reduces not only mortality but also other cardiac events. Coumadin has an even more impressive record. It reduces mortality by 25 to 35% and recurrent infarctions by 50%, with an acceptable risk for cerebral hemorrhage (1 to 2% per year). After coronary revascularization, aspirin is the therapy of choice for both coronary angioplasty and bypass surgery, with a few exceptions (poor left-ventricular function, stents in small arteries, etc.). Ticlopidine seems to be a valid alternative. Artificial heart valves require coumadin, with the exception of bioprosthesis in patients without additional risk factors. The same holds true for atrial fibrillation, except for the lone variety in young patients. Every cardiologist has to be cognizant of the armamentarium of anticoagulants and thoroughly trained in their indications and handling. PMID- 7482378 TI - [Erythropoietin, a milestone in the history of nephrology]. AB - Erythropoietin, a glycoprotein, is synthesized mainly in the kidney. With the destruction of renal tissue, erythropoietin production decreases; this is a major factor in the development of anemia in patients with renal failure. For about ten years now, recombinant human erythropoietin has been available for the treatment of renal anemia. All patients with renal insufficiency, independent of their plan for future renal replacement therapy, may benefit from erythropoietin. At what extent of anemia erythropoietin therapy should be started is still discussed and is certainly dependent on the degree of the patient's impairment by his anemia. Before beginning a therapy with erythropoietin, other forms of anemia observed in patients with renal failure, i.e. mainly iron deficiency, have to be excluded. A strict monitoring of hematocrit during treatment with erythropoietin is mandatory. Hypertension, seizures and cardiovascular complications have been observed with overdosing of erythropoietin. Special emphasis of this review is therefore put on the discussion of the dynamics of the erythropoietin-red cell system. PMID- 7482379 TI - [Growth hormone: who can benefit from it?]. AB - Recombinant human growth hormone (GH) has become available in unlimited quantities. Classic indications for GH therapy include GH deficiency of childhood and adult onset, Turner syndrome and chronic renal insufficiency. Nonclassic indications for GH use are short stature without GH deficiency, catabolic states, severe burns, osteoporosis. Controlled studies are necessary to prove a potential benefit of GH in nonclassic or new situations. The prescription of GH should be supervised by specialists to ensure that this expensive drug is used judiciously. PMID- 7482380 TI - [Bile acids in liver diseases--current indications]. AB - Ursodeoxycholic acid (UDCA) has beneficial effects in cholestatic diseases such as primary biliary cirrhosis, primary sclerosing cholangitis and cholestasis of pregnancy. In chronic hepatitis and alcoholic hepatitis, an effect of UDCA is uncertain. After organ transplantation (liver, bone, heart), favorable effects of UDCA which await confirmation were observed. UDCA very likely has a beneficial effect in children with Byler's and Alagille's syndrome, extrahepatic biliary atresia after the Kasai procedure and in cholestasis of cystic fibrosis. Unclear is the effect of UDCA in benign intermittent cholestasis (Summerskill-Tygstrup syndrome). Children with cholestasis and inborn errors of bile acid synthesis need additional administration of a primary bile acid (cholic acid, chenodeoxycholic acid). UDCA treatment in general does not lead to definitive cure of the disease but to improvement of laboratory parameters and possibly of symptoms and liver histology, and liver transplantation may be postponed to a later time point. PMID- 7482381 TI - [Carnitine: vitamin or doping?]. AB - Carnitine (L-trimethyl-3-hydroxy-4-ammonio-butyrate) is a chemically simple substance with essential functions in energy metabolism. Most of the carnitine needed is ingested by the diet, in particular by meat and dairy products. The rest is biosynthesized, starting from the amino acids lysine and methionine. In humans, carnitine biosynthesis is just sufficient to avoid symptomatic carnitine deficiency, when carnitine intake by the diet is absent or low. Carnitine can therefore not be regarded as a vitamin. Primary carnitine deficiency is mostly due to increased renal carnitine excretion because of a defective carnitine transport in the proximal tubule. Secondary carnitine deficiency can develop due to increased excretion of acylcarnitines in patients with organic acidurias or after longterm treatment with pivaloyl-esters or other drugs. Symptomatic carnitine deficiency should be treated with pharmacological doses of carnitine. There are no studies supporting the concept that acute administration of carnitine is associated with increased physical performance in athletes. Acute ingestion of carnitine by athletes is not doping, but cannot be advised from a scientific standpoint. So far, there are no randomized controlled studies investigating the effect of chronic carnitine administration on physical performance in athletes. PMID- 7482382 TI - [Sex hormone replacement following menopause--for all women? A plea for prevention using estrogens and gestagens]. AB - After the menopause, a growing proportion of women will have a good chance to add three decades or more to their lifetime. They must decide whether to start long term hormonal replacement therapy or to accept the risks of osteoporosis, fracture, cardiovascular disease, and a variety of psychological and physical problems as 'natural' destiny. The syndrome of postmenopausal endocrine deficiency is a primary glandular insufficiency, which in principle requires substitution with the secretory product of the gland. Postmenopausal osteoporosis and fractures are consequences of a pathological dysfunction of calcium metabolism. After estrogen withdrawal, the impaired hepatic and renal synthesis of calcitriol will result in a reduced intestinal resorption of calcium. Parathyroid hormone may initiate a vicious circle by acceleration of bone resorption, mobilization of bone calcium and a tendency to hypercalcemia. The failure to preserve circulating calcium due to 'escape' from calcitonin and a decreased renal tubular back-resorption are followed by an increased loss of calcium from a 'renal calcium leak', resulting in hypercalcuria. In order to maintain homeostasis, additional calcium is required, which will be supplied from accelerated bone resorption. Thus, the circle is closed by renewed osteolysis. The process is associated with accelerated bone turnover and a negative balance of calcium and bone. After a variable time interval which depends on the individual bone mass and rate of bone loss, these events will inevitably result in osteoporosis. Estrogen replacement will interrupt the circle, decelerate bone turnover, and re-establish a positive balance of calcium and bone. Estrogen withdrawal also favors an 'atherogenic' set of lipoproteins, which is strongly associated with increased coronary risk. Substitution with estrogens will favor a 'protective' profile of lipoproteins and cut the cardiovascular risk to about one half. Progestogens, in particular the 17 alpha-alkylated 19-nor-steroids, may reduce the favorable effects of estrogens on lipoproteins in a dose-dependent manner; however, they do not impair the antiatherogenic estrogen effects, even in presence of an atherogenic profile of lipoproteins. Thus, these anti-estrogenic effects of progestogens, at least in nonhuman primates, do not have clinical significance.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 7482384 TI - Provocative agents in panic disorder. AB - The pharmacological challenge strategy involves giving a provoking agent under controlled rules to clarify some aspect of behavioural or biological function. Various agents such as sodium lactate, carbon dioxide, caffeine, yohimbine, isoprenaline and now cholecystokinin have been used as provoking agents in healthy volunteers as well as in panic patients. Results obtained in this field are updated, with emphasis on the potential mechanisms of action. It is concluded that there may be a final pathway between carbon dioxide, sodium lactate, and cholecystokinin inducing panic attacks. PMID- 7482383 TI - [Therapie 1946-1995: 50 years of publication]. AB - Therapy is celebrating its 50th anniversary with a completely new layout and a special edition that will be released at the first meeting of the European Association for Clinical Pharmacology and Therapeutics. This is also an appropriate time to look at its history. Immediately after the Second World War, in 1946, the Societe de Therapeutique, which was originally founded in 1866, was resurrected as the Societe Francaise de Therapeutique et de Pharmacodynamie. This society launched Therapie. After a rather difficult beginning, the society began to grow and its journal appeared regularly. There were 1000 society members in 1961 and the journal published 1000 pages, mostly devoted to experimental pharmacology. The society began to concentrate on human pharmacology from 1972, and the number of articles on therapeutics and clinical pharmacology published in THERAPIE increased each year. The section on animal pharmacology was dropped in 1981 and the change in the journal's coverage was complete. The name of the society was changed to the 'Societe Francaise de Therapeutique et de Pharmacologie Clinique'. THERAPIE became, and has remained, the only French medical journal entirely devoted to clinical pharmacology and therapeutics. PMID- 7482385 TI - Single dose of prednisone does not induce amphetamine-like subjective effects in healthy subjects. AB - Among the methods developed in assessing abuse liability, the behavioural and subjective effects of drugs can be recorded using the Addiction Research Center Inventory (ARCI) in drug-experienced subjects and normal volunteers. Sixteen healthy volunteers with no history of drug abuse participated in the study. The subjective, behavioural and physiological effects of prednisone (30 and 60 mg) were compared with those of dextroamphetamine (15 mg) and placebo in a randomized double-blind Latin square design. The self-questionnaires (ARCI, Profile of Mood States, Visual Analogue Scales and Sleep Questionnaire) were completed before, 1, 2, 4 and 8 h post single oral dosing. Results showed that subjective effects of the two studied doses of prednisone did not resemble those induced by dextroamphetamine (15 mg). These results indicate that oral single doses of prednisone do not possess amphetamine-like subjective effects in a healthy population. The well established psychostimulant effect of amphetamine have been replicated on almost all subjective assessments. PMID- 7482386 TI - [Comparison of blood pressure profile during exercise with benazepril 10 mg and benazepril 5 mg/hydrochlorothiazide 6.25 mg fixed combination]. AB - This double-blind placebo-controlled study was designed to compare the efficacy of benazepril 10 mg (BZ 10) and the low-dose combination benazepril 5 mg/hydrochlorothiazide 6.25 mg (BZ + HCTZ) on exercise blood pressure in 71 patients with mild to moderate hypertension (diastolic blood pressure 95 to 114 mmHg at the end of a 2 week placebo run-in). A significant fall of systolic (SBF) and diastolic (DBP) blood pressure at rest was seen after a 4-week treatment period in the two active groups (-21.9/-10.3 mmHg with BZ 10, -19.5/-11.2 mmHg with BZ + HCTZ) and was greater in these two groups than in the placebo group ( 7.5/-2.5 mmHg, p < 0.01). For each level of the exercise test on bicycle ergometer, SBP falls seen after 1 month were significant in the two active groups (from -15 to -23 mmHg depending on the level and treatment group). The evolution of the SBP/heart rate slope was significantly different among the two active groups in favour of the BZ 10 regimen. The exercise test proved to be a valuable tool to show any difference between two treatments with comparable efficacy on SBP/DBP at rest. PMID- 7482388 TI - Critical reading of the meta-analysis of clinical trials. AB - In this paper we shall present the general principles of meta-analysis and will then discuss the various factors needed to evaluate a meta-analysis: description of the problem; definition of the outcome(s) (primary and secondary); methods for identifying and selecting trials for inclusion; statistical methods used; and the presentation and discussion of the results. We shall then examine other problems such as the detection of bias, the validity of the information provided by the meta-analysis, the problem of heterogeneity, the sensitivity and robustness of the meta-analysis, quality criteria for a meta-analysis, and how to locate published meta-analyses. Finally we present a decision algorithm which should help answer the question: should and can the results from the meta-analysis be integrated into clinical practice? PMID- 7482387 TI - Excitatory amino acid receptors and neurodegeneration. AB - This review describes recent advances in our understanding of the pharmacology of excitatory amino acid receptors, and the application of this knowledge to the unravelling of the aetiology of neurodegenerative diseases, and to their therapy. Ionotropic excitatory amino acid receptors can be divided into two large families, the NMDA receptor family, and the AMPA/kainate receptor family. Receptor cloning studies have shown there to be a large number of potential subtypes of receptors in both these families. Antagonists have been developed for the NMDA receptor which can interact with at least four independent drug recognition sites on the receptor. For the AMPA/kainate receptor, two classes of antagonist have so far been identified. Reasonably potent, selective and brain penetrating antagonists now exist for virtually all these sites, and compounds inhibiting the release of glutamic acid presynaptically have also been identified, such as riluzole. The ability of glutamic acid to kill neurons (excitotoxicity) seems to be mediated, in most cases, by an interaction with NMDA receptors, leading to an uncontrollable rise in intracellular calcium concentrations and thence cell lysis and death. The setting-up of glutamatergic loops seems to be a key process in the maintenance, spread and amplification of neurodegenerative foci. The existence of such processes has been amply demonstrated in animal models of stroke, in which both NMDA and AMPA/kainate receptor antagonists have neuroprotective effects. Clinical trials are underway with NMDA receptor antagonists in stroke. Excitotoxic mechanisms probably also contribute to pathology in head trauma and viral encephalopathy. Ingestion of excitatory amino acids may play a role in neurological conditions of dietary aetiology, such as neurolathyrism and domoic acid intoxication. For chronic neurodegenerative diseases, the role of excitatory amino acids is much less clear, although there is some evidence for the existence of excitotoxic mechanisms in amyotrophic lateral sclerosis. Evidence from animal models suggests that drugs that block glutamatergic neurotransmission might be beneficial in Parkinson's disease, Huntington's chorea and amyotrophic lateral sclerosis, but the relevance of these animal models to the human pathology is not clear. However, preliminary clinical results suggest riluzole to be efficacious in prolonging survival in amyotrophic lateral sclerosis, and certain weak NMDA receptor antagonists are currently used in the treatment of Parkinson's disease. The next few years could witness a breakthrough in the treatment of neurological conditions as drugs that interfere with glutamatergic transmission become available for clinical use. PMID- 7482389 TI - INDANA: a meta-analysis on individual patient data in hypertension. Protocol and preliminary results. AB - The overall effect of antihypertensive drug treatment has been well documented. The proportion of patients who benefit varies according to their baseline cardiovascular risk, and is small for the majority of people treated. Some investigators propose limiting the treatment target population to patients at high cardiovascular risk, but several assumptions must be made to justify this procedure. The INDANA project is a meta-analysis based on individual patient data, and thus offers the opportunity to check the validity of these assumptions. Its main objective is to identify responders (and non-responders) in the drug treatment of hypertension. The rationale and methods for such an approach are presented here, with the solution for some technical problems. The conclusion of the data collection has shown that the project is feasible. The results of the main analysis should be available in 1996, and should contribute to the selection of responders and to the individualization of the treatment of hypertension. PMID- 7482390 TI - [Women in clinical trials]. AB - In a workshop held in Giens in September 1994, representatives of drug companies and scientists met to discuss the place of women in clinical trials. They recommend that, in phase II and III studies, men and women should be included in a proportion equivalent to that observed for the condition studied. They also recommend that the impact of the gender on the results should be systematically studied. PMID- 7482391 TI - [Gene therapy]. AB - This paper considers the prerequisites for gene therapy. This new field of therapeutics has not yet proved its efficacity nor its safety. Use, as a drug of DNA molecules which can modify host genome and environment as well requires specific prerequisites; among them, animal experimental models are essential despite their limitations. The different preclinical and clinical steps in the design of gene therapy studies are also discussed. PMID- 7482392 TI - [The future of pharmacological models]. AB - Do pharmacological models have a future? This was the question that had to be answered during seminar n.3 of the annual clinical pharmacology meeting in GIENS. The concept of 'model' is very extensive: it comprises both simple physiological testing and the replication in animals of human diseases. The main problems of pharmacological models are their predictive value and their validity in relation to the pragmatic target of finding new active molecules. Among numerous models proposed by the participants, three types have been selected as examples in this paper: a human model (cholecystokinin inducing panic attacks), the goal of which is to discover new molecules active in panic disorders. an animal model close to clinical features (coronary restenosis) which was to date unable to help in identifying molecules acting in human pathology transgenic animals as tools in drug development. The guidelines are very clear: models, however far they are from human pathology, are useful in predicting new molecular developments. Models are necessary steps to go from receptors to ill patients. PMID- 7482393 TI - [Determination of drug posology in pediatrics]. AB - The dosage of drugs which might be used in children must be determined to avoid empirical use, even when no application has been submitted for a paediatric licence. Toxicological evaluation and assessment of the effects on growth, an adapted pharmaceutical form and paediatric pharmacokinetic and adult clinical data are essential before conducting trials designed to determine the paediatric dosage. The dose used during preliminary studies is extrapolated from the adult dose expressed in relation to weight, tested in a dose-effect study, and then more accurately defined on the basis of pharmacokinetic data in different age groups. Obtaining consent from both parents for studies whose direct benefit is not always obvious, as well as the global cost of these studies, constitute drawbacks to paediatric drug development. Incentives to determine a paediatric dosage could consist of public participation in funding, prolongation of the patent, and granting an advantageous price for a specifically paediatric pharmaceutical form or indication. PMID- 7482394 TI - [Arterial wall: a new pharmacological and therapeutic target?]. AB - In recent years, endothelial dysfunction and arterial remodelling in various cardiovascular diseases have emerged as two key concepts, with numerous interrelationships. Both endothelial dysfunction and arterial remodelling occur in various pathologies including heart failure, atherosclerosis, restenosis after angioplasty, and pulmonary hypertension, and have modified the therapeutic approach by offering new pharmacological targets: specific receptors not only at the site of the vascular smooth muscle cells but also on the endothelial cells, growth factors that stimulate proliferation of smooth muscle, and receptors and enzymes of the extracellular matrix. Among the various substances under research, the present review will discuss angiotensin II receptor antagonists, endothelin receptor antagonists, nitrates-NO donors, potassium channel activators, and substances interfering with proteoglycans and other components of the extracellular matrix. PMID- 7482395 TI - Resistance to activate protein C, the Arg506 to Gln mutation in the factor V gene, and venous thrombosis. Functional tests and DNA-based assays, pros and cons. PMID- 7482396 TI - Homozygous APC-resistance combined with inherited type I protein S deficiency in a young boy with severe thrombotic disease. AB - Inherited resistance to activated protein C (APC) is a frequent cause of familial thrombosis. It is associated with a factor V gene point mutation replacing arginine506 in the APC-cleavage site with a glutamine. Thrombotic events are rare during childhood even in patients with homozygous APC-resistance. We now wish to report on a case of severe venous thrombosis, in a 10-year-old boy. He was found to have pronounced APC-resistance due to homozygous factor V gene mutation in combination with inherited type I protein S deficiency. The two traits were independently inherited in the family. The APC-resistance was partially corrected by adding factor V, whereas added protein S was without effect. This is the first reported case of homozygous APC-resistance combined with another inherited prothrombotic disorder. It illustrates how multiple genetic defects may provoke thrombosis at young age and emphasizes the need of complete evaluation of thrombotic patients in order to determine whether multiple risk factors exist. PMID- 7482397 TI - Protein S mRNA in patients with protein S deficiency. AB - A protein S gene polymorphism, detectable by restriction analysis (BstXI) of amplified exonic sequences (exon 15), was studied in seven Italian families with protein S deficiency. In the 17 individuals heterozygous for the polymorphism the study was extended to platelet mRNA through reverse transcription, amplification and densitometric analysis. mRNA produced by the putative defective protein S genes was absent in three families and reduced to a different extent (as expressed by altered allelic ratios) in four families. The allelic ratios helped to distinguish total protein S deficiency (type I) for free protein S deficiency (type IIa) in families with equivocal phenotypes. This study indicates that the study of platelet mRNA, in association with phenotypic analysis based upon protein S assays in plasma, helps to classify patients with protein S deficiency. PMID- 7482398 TI - Identification of eight point mutations in protein S deficiency type I--analysis of 15 pedigrees. AB - We described molecular genetic studies of 15 patients with protein S deficiency type I (i.e. reduced total protein S antigen). All the exons of the PROS 1 gene were analyzed both by PCR and direct sequencing in all 15 probands. This analysis led to the identification of point mutations affecting eight individuals. One of these mutations (codon-25, insertion of T) has been described previously in a Dutch pedigree. The other mutations are novel and all are located in exons that code for the protein S domain that is homologous to the steroid hormone binding globulins. They include two amino acid replacements (one individual with 340 Gly- > Val, and two individuals with 467 Val --> Gly), and four frameshift mutations due to either one bp deletions (in codon 261 deletion of T and in codon 267 deletion of G) or insertions (in codon 565 insertion T and after codon 578 insertions of C). Studies performed in six families (totalling 43 subjects) showed cosegregation of the genetic abnormality with reduced plasma protein S levels, and provided genetic evidence for a heterozygous protein S deficiency in 25 of them. The yield of mutations in this study (53%) confirms that the percentage of protein S deficient cases in which a point mutation is found remains low. PMID- 7482399 TI - Abnormal processing of the glycoprotein IIb transcript due to a nonsense mutation in exon 17 associated with Glanzmann's thrombasthenia. AB - We analyzed the molecular genetic defect responsible for type I Glanzmann's thrombasthenia in a Japanese patient. In an immunoblot assay using polyclonal anti-GpIIb-IIIa antibodies, some GPIIIa (15% of normal amount) could be detected in the patient's platelets, whereas GPIIb could not (< 2% of normal amount). Nucleotide sequence analysis of platelet GPIIb mRNA-derived polymerase chain reaction (PCR) products revealed that patient's GPIIb cDNA had a 75-bp deletion in the 3' boundary of exon 17 resulting in an in-frame deletion of 25 amino acids. DNA analysis and family study revealed that the patient was a compound heterozygote of two GPIIb gene defects. One allele derived from her father was not expressed in platelets, and the other allele derived from her mother had a 9644C--> T mutation which was located at the position -3 of the splice donor junction of exon 17 and resulted in a termination codon (TGA). Moreover, quantitative analysis demonstrated that the amount of the abnormal GPIIb transcript in the patient's platelets was markedly reduced. Thus, the C --> T mutation resulting in the abnormal splicing of GPIIb transcript and the reduction in its amount is responsible for Glanzmann's thrombasthenia. PMID- 7482400 TI - Marked increase of activated factor VII in uremic patients. AB - We investigated plasma activated factor VII (FVIIa) levels in uremic patients (nondialysis group: n = 38; dialysis group: n = 36) and healthy controls (n = 32). We also measured the plasma levels of thrombomodulin (an indicator of endothelial cell injury) and tissue factor. Plasma FVIIa showed a marked increase in the nondialysis group (mean [95% confidence interval]: 4.6 [4.1-5.1] ng/ml, p < 0.0001) with the progressive impairment of renal function, as indicated by the serum creatinine level, when compared with the 32 controls (2.8 [2.5-3.1] ng/ml), and was further increased in the dialysis group (6.1 [5.5-6.8] ng/ml, p < 0.001 vs. nondialysis group). Plasma levels of thrombomodulin and tissue factor were also higher in the nondialysis group than the control group, and were further increased in the dialysis group. Plasma tissue factor levels did not show any correlation with FVIIa or thrombomodulin in both the nondialysis and dialysis groups. Thus, circulating tissue factor appears to be released by a different mechanism from thrombomodulin and may not contribute to the direct activation of factor VII in uremic patients. On the other hand, the plasma level of thrombomodulin was positively correlated with that of FVIIa in the nondialysis group, and this correlation was independent of renal function. Thus, enhanced conversion of factor VII zymogen to FVIIa, probably related to endothelial cell injury, may be a risk factor for cardiovascular events in uremic patients. PMID- 7482402 TI - Detection of ten new mutations by screening the gene encoding factor IX of Danish hemophilia B patients. AB - Hemophilia B is caused by a wide range of mutations. In order to characterize the mutations among patients in Denmark, we have systematically screened the entire coding region, the promoter region and exon flanking sequences of the gene encoding factor IX using single strand conformation and heteroduplex analyses. Patients from 32 different families were examined, and point mutations (23 different) were found in all of them. Ten of the mutations have not been reported by others; they include a splice site mutation, a single base pair deletion, and missense mutations. Notably, the study contains a female patient and a previously described Leyden mutation. In ten families with sporadic cases of hemophilia B, all 10 mothers were found to be carriers. The origin of two of these mutations was established. PMID- 7482401 TI - Antiphospholipid antibodies, haemostatic variables and thrombosis--a survey of 144 patients. AB - Several clotting abnormalities have been put forth to explain the thrombotic tendency of the antiphospholipid syndrome, but a possible role for fibrinogen and von Willebrand factor has been poorly investigated. The present cross-sectional retrospective study evaluated the relationship of IgG anticardiolipin antibodies, lupus anticoagulants, fibrinogen and von Willebrand factor with the occurrence of arterial and venous thromboses in patients with antiphospholipid antibodies. Among the clotting assays for the detection of lupus anticoagulant, dilute Russell's viper venom time correlated with a history of venous thrombosis more strongly than activated partial thromboplastin time (p < 0.0002 vs p < 0.009) and was the only test which correlated with a history of arterial thrombosis (p < 0.01), also at low levels of IgG anticardiolipin antibodies (p = 0.003). By regression analysis, and after correction for confounders, serum levels of IgG anticardiolipin antibodies were found to be positively associated with the number of venous events (p < 0.001). Plasma levels of fibrinogen and von Willebrand factor were associated with each other (p < 0.0001; r: 0.48) and with the occurrence of arterial and venous thromboses (p < 0.001). Moreover, plasma levels of fibrinogen and von Willebrand factor in thrombotic patients with antiphospholipid antibodies were significantly higher than those of a control group of thrombotic patients who suffered thrombosis for other reasons (p < 0.0001 and p = 0.0008 respectively). Titres of IgG anticardiolipin antibodies correlated with plasma levels of von Willebrand factor (p < 0.0001; r: 0.42).(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7482403 TI - Variability of in vivo recovery of factor IX after infusion of monoclonal antibody purified factor IX concentrates in patients with hemophilia B. The Mononine Study Group. AB - Monoclonal antibody purified factor IX concentrate, Mononine (Armour Pharmaceutical Company, Kankakee, Illinois, USA), is a recently developed replacement factor concentrate for the treatment of patients with hemophilia B. The pharmacokinetic properties of monoclonal antibody purified factor IX concentrate (MAb Factor IX concentrate) have been evaluated in only small samples of patients, and little is known about those factors that might influenced in vivo recovery of factor IX after infusion is a larger patient population. In vivo recovery of factor IX was therefore evaluated for 80 different indications in 72 patients who received MAb Factor IX concentrate for the management of spontaneous or trauma-induced bleeding, or as prophylaxis with surgery. The average recovery after infusions for presurgical pharmacokinetic analysis (mean +/- standard deviation) was 1.28 +/- 0.56 U/dl rise per U/kg infused (range 0.41-2.80), and the average recovery after all infusions for treatment was 1.23 +/- 0.49 U/dl rise per U/kg infused (range - 0.35-2.92). Recovery values for multiple MAb Factor IX doses in a given patient were also variable; the average recovery was 1.22 +/- 0.53 U/dl rise per U/kg given, and standard deviations ranged from 0.03 to 1.26. Patient age, weight, and MAb Factor IX concentrate dose minimally but significantly influenced factor IX recovery. There was no significant effect of either race, history of previous thrombotic complications during treatment with other replacement factor concentrates, or bleeding state on recovery. All of the patients treated with this preparation experienced excellent hemostasis, and no thrombotic complications were observed. PMID- 7482404 TI - The thrombin activation pathway modulates the assembly, structure and lysis of human plasma clots in vitro. AB - Thrombin activation of the soluble plasma protein fibrinogen is vital for successful haemostasis. Thrombin is generated from prothrombin by the prothrombinase complex which also includes factor Xa, factor Va, Ca2+ and a procoagulant membrane surface. Factor X activation is catalysed in a complex including either factor VIIa and tissue factor, or factor IXa and factor VIIIa. Factor IXa can be generated either by the factor VIIa/tissue factor complex or by factor XIa which is in turn produced by the contact phase reactions in vitro. Once activated, fibrinogen develops into the fibrin polymeric matrix at the site of injury. It is not known to what extent the properties of this haemostatic plug are sensitive to the pathway leading up to thrombin generation. Here static human plasma is studied in vitro using magnetically induced birefringence. It is shown that the contact phase/factor XIa pathway gives rise to linear fibrin assembly process curves whereas the factor VIIa/ tissue factor activation of factor X provokes largely sigmoid assembly. The latter pathway also causes the formation of significantly thicker fibres even though assembly is more rapid. This result is the inverse of that anticipated from the study of simple model systems. Whilst the streptokinase activated lysis both types of clot exhibits similar biphasic kinetics, an exponential main phase followed by a sigmoidal tailing off, the data suggest that clots produced by the contact phase/factor XIa pathway are more recalcitrant to lysis. These results demonstrate that the profile of thrombin generation not only determines the kinetics of assembly but also influences the rate of lysis and structure of the haemostatic plug. PMID- 7482405 TI - Cellular localization of enzymatically active thrombin in intact human tissues by hirudin binding. AB - Cellular sites of coagulation activation within complex, intact tissues have been studied by immunohistochemical techniques. Hirudin, a specific and high affinity inhibitor of the active site of thrombin, together with antibody to hirudin were applied to sections of AMeX-fixed specimens of normal lung, kidney, placenta, freshly incised skin and unperturbed skin obtained at fresh autopsy; to rheumatoid synovial tissue; and to malignant tissue from a variety of tumor types. Staining for thrombin was observed selectively on pulmonary alveolar, rheumatoid synovial, and placental macrophages that express an intact extrinsic coagulation pathway. Staining was also observed restricted to the endothelium of capillaries in freshly incised skin but not in either unperturbed skin or in aged incisions. Staining of tumor cell bodies was observed in small cell carcinoma of the lung, renal cell carcinoma, and malignant melanoma tissues that we found previously to show tumor cell-associated procoagulant activity. This staining occurred commonly on cells within the tumor mass that were distant from stromal fibrinogen/fibrin. By contrast, tumor-associated macrophage but not tumor cell staining was seen in adenocarcinoma and squamous cell carcinoma of the lung, and little or no staining was seen colon cancer tissue. Negative controls in which either the hirudin probe or its antibody were omitted failed to show staining. These results are in accord with previous findings and suggest that such techniques may be useful for studying the cellular sites of thrombin generation in intact tissues. We postulate that administration of potent and specific thrombin antagonists, such as hirudin, to patients with relevant tumor types might be followed by homing of hirudin to tumor cells in vivo so that effects of local thrombin generation on malignant progression can be determined. PMID- 7482406 TI - Synchronized inhibition of the phospholipid mediated autoactivation of factor XII in plasma by beta 2-glycoprotein I and anti-beta 2-glycoprotein I. AB - Lupus anticoagulants are a group of antibodies commonly found in patients with autoimmune diseases such as systemic lupus erythematosus. Lupus anticoagulants inhibit phospholipid dependent coagulation and may bind to negatively charged phospholipids. Recent studies have suggested an association between anti-beta 2 glycoprotein I and a lupus anticoagulant, whose activity is frequently dependent on the presence of beta 2-glycoprotein I. Based on these observations, the effect of anti-beta 2-glycoprotein I on the autoactivation of factor XII in plasma was investigated. Autoactivation initiated by the presence of negatively charged phospholipids, but not by sulfatide, was strongly inhibited by immunoaffinity purified anti-beta 2-glycoprotein I. The dose-response curve of anti-beta 2 glycoprotein I was identical with that of a precipitating antibody, showing no inhibition at low and high antibody dilutions and maximal inhibition at an intermediate dilution. At high antibody concentrations, an increased rate of factor XIIa activation was observed. This increase was of the same magnitude as the decreased rate observed in plasma supplemented with the same amount of beta 2 glycoprotein I as in the plasma itself. This confirms the inhibitory effect of beta 2-GP-I on the contact activation and shows that inhibition is effective on the autoactivation of factor XII in plasma. The inhibitory action of beta 2 glycoprotein I was independent of the inhibition caused by the anti-beta 2 glycoprotein I/beta 2 glycoprotein I complex suggesting a synchronized inhibition of factor XII autoactivation by beta 2-glycoprotein I and anti-beta 2 glycoprotein I. The inhibition caused by the antibody is suggested to be caused by a reduced availability of negatively charged phospholipids due to the binding of the anti-beta 2-GP-I/beta 2-GP-I complex. This complex may be a lupus anticoagulant. PMID- 7482407 TI - Soluble thrombomodulin purified from human urine exhibits a potent anticoagulant effect in vitro and in vivo. AB - We examined the anticoagulant activity of two major molecules of soluble thrombomodulin purified from human urine. The apparent molecular weights of these urinary thrombomodulins (UTMs) were 72,000 and 79,000, respectively. Both UTMs showed more potent cofactor activity for protein C activation [specific activity > 5,000 thrombomodulin units (TMU)/mg] than human placental thrombomodulin (2,180 TMU/mg) and rabbit lung thrombomodulin (1,980 TMU/mg). The UTMs prolonged thrombin-induced fibrinogen clotting time (> 1 TMU /ml), APTT (> 5 TMU/ml), TT (> 5 TMU/ml) and PT (> 40 TMU/ ml) in a dose-dependent fashion. These effects appeared in the concentration range of soluble thrombomodulins present in human plasma and urine. In the rat DIC model induced by thromboplastin, administration of UTMs by infusion (300-3,000 TMU/kg) restored the hematological abnormalities derived from DIC in a dose-dependent fashion. These results demonstrate that UTMs exhibit potent anticoagulant and antithrombotic activities, and could play a physiologically important role in microcirculation. PMID- 7482408 TI - Transforming growth factor beta 1 and beta 2 induce down-modulation of thrombomodulin in human umbilical vein endothelial cells. AB - To investigate the effects of transforming growth factor-betas (TGF-betas) on endothelial anticoagulant activity, we assayed thrombomodulin (TM) activity and antigen levels of human umbilical vein endothelial cells (HUVECs) incubated with TGF-betas in vitro. TGF-beta 1 suppressed surface TM activity and surface TM antigen levels maximally 12 h after incubation in dose-dependent manners. TGF beta 2 was almost equipotent with TGF-beta 1 for the suppression of them. Both TGF-betas suppressed total TM antigen level in HUVECs, and the time course of the suppression was similar to that of the cell surface TM antigen level. The maximal reductions of TM mRNA levels by TGF-betas were observed at several hours ahead of those observed in both surface and total TM antigens levels, suggesting that the TGF-beta-mediated suppression of TM antigen of HUVECs is primarily regulated at the TM mRNA level. Our present work suggests that the down-modulation of TM level induced by TGF-betas in HUVECs contributes in vivo to promoting the thrombogenesis either at the sites of injury of vessel walls, such as atherosclerotic lesions where TGF-beta 1 is released from platelets, smooth muscle cells and monocytes, or at neovascular walls in tumors secreting TGF-beta 2. PMID- 7482409 TI - IL-6 upregulates protein S expression in the HepG-2 hepatoma cells. AB - Several pro-inflammatory cytokines have been shown to be important in the modulation of the procoagulant response. However, what role these cytokines may have in the regulation of coagulation inhibitors is poorly understood. While the hepatocyte is a primary site of synthesis for the anticoagulant protein C and S, it is also a major target cell for the proinflammatory cytokine, IL-6. We have found that stimulation of HepG-2 hepatoma cells with IL-6 (5 ng/ ml) significantly increased the production of the anticoagulant cofactor, protein S, in both a time and dose dependent fashion. This increase was seen at both the RNA and protein level. A mouse monoclonal neutralizing antibody to human IL-6 suppressed the IL-6 effect in a concentration dependent fashion. IL-6 also increased the release of the C4b-binding protein but had no effect on protein C production. When combined with either dexamethasone or soluble IL-6 receptor, the IL-6 response was significantly enhanced. Oncostatin M, a functionally related cytokine, had a similar effect while other related cytokines, IL-11 and leukemia inhibitory factor, only had a marginal effect. IL-1, TGF-beta, and TNF-alpha had no significant effect on protein S production. These results indicate that IL-6 may play an important regulatory role in the anti-coagulant pathway. PMID- 7482410 TI - Protection of single-chain urokinase-type plasminogen activator (scu-PA) in aprotinin treated cardiac surgical patients undergoing cardiopulmonary bypass. AB - Intraoperative high-dose aprotinin administration has been shown to reduce the intra-and postoperative blood loss in cardiac surgery. The haemostatic effect has been attributed to platelet preserving properties and to inhibition of contact activation reducing thrombotic and fibrinolytic activity during and after cardiopulmonary bypass (CPB). Here we report on the effects of aprotinin on urokinase-type plasminogen activator, especially on the protection of the zymogen single-chain urokinase-type plasminogen activator (scu-PA). scu-PA occurs cell associated as well as free in the circulation (concentration 50 pM, half-life 5 min), and is potentially activated by kallikrein and plasmin, both potent targets for aprotinin. The generated active two-chain u-PA (tcu-PA) is a powerful activator of fibrinolysis. Sixteen male patients undergoing myocardial revascularization were randomly assigned to aprotinin treatment (A) or control group (C). Plasma concentration of total u-PA antigen and of the specific forms scu-PA(zymogen) and tcu-PA(active enzyme) were measured at different stages intraoperatively and two hours postoperatively. After an initial drop due to haemodilution at the onset of CPB, the concentrations of circulating u-PA forms restored intraoperatively in A, but remained subnormal in C until the end of the observation period. The concentration of total u-PA antigen of shed mediastinal blood was both in A and C two-fold higher than in the circulation, but the antigen was preserved as the zymogen scu-PA in A and largely converted to an inactive, non activatable form in C. Intra- and postoperative blood losses were less than half the amount in A as compared to C.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7482411 TI - Localization of a vitronectin binding region of plasminogen activator inhibitor 1. AB - The PAI-1 binding site for VN was studied using two independent methods. PAI-1 was cleaved by Staph V8 protease, producing 8 fragments, only 2 of which bound to [125I]-VN. These fragments were predicted to overlap between residues 91-130. Since PAI-2 has structural homology to PAI-1, but does not bind to vitronectin, chimeras of PAI-1 and PAI-2 were constructed. Four chimeras, containing PAI-1 residues 1-70, 1-105, 1-114, and 1-167 were constructed and expressed in vitro. PAI-1, PAI-2, and all of the chimeras retained inhibitory activity for t-PA, but only the chimera containing PAI-1 residues 1-167 formed a complex with VN. Together, these results predict that the VN binding site of PAI-1 is between residues 115-130. PMID- 7482412 TI - Plasminogen activator inhibitor type-1 determines plasmin formation in patients with ischaemic heart disease. AB - The aim of the present study was to find out whether plasminogen activator inhibitor type-1 (PAI-1) controls the formation of plasmin in patients with ischaemic heart disease. We examined PAI activity, PAI-1 antigen, tissue type plasminogen activator (t-PA) activity, t-PA antigen, plasmin-alpha2-antiplasmin complex (PAP-complex) and fibrin degradation products D-dimer in 62 patients before (unstimulated) and after infusion of 1-desamino-8-D-arginine vasopressin (DDAVP; stimulated). DDAVP was used in a standardized dose to trigger the release of t-PA from the vascular endothelium. We observed that under basal conditions (unstimulated) median plasma t-PA activity for the whole group of patients was 86.5 mIU/ml (0-900), and after stimulation 2550 mIU/ml (0-6800), P < 0.0001; median plasma concentration of t-PA antigen was 14.7 ng/ml (7.0-115.5) under basal conditions, and after stimulation 34.1 ng/ml (15.8-58.6), P < 0.0001; median plasma PAI activity was 16.9 IU/ml (1.5-144.8) under basal conditions, and after stimulation 3.1 IU/ml (0-118.5), P < 0.0001; median plasma concentration of PAI-1 antigen was 21.5 ng/ml (8.1-132.2) under basal conditions, and after stimulation 14.9 ng/ml (4.8-149.0), P < 0.0001; the median plasma concentration of PAP-complex was 469.5 ng/ml (185.0-1802.0) under basal conditions, and after stimulation 695.5 (243.0-2292.0), P < 0.0001; median plasma concentration of D dimer was 298.0 ng/ml (103.0-948.0) under basal conditions, and after stimulation 296.5 ng/ml (97.0-917.0), P < 0.0008.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7482413 TI - Enhanced vascular plasminogen activator (t-PA) release by epinephrine in aged rats. AB - Adrenergic stimulation induces an acute fibrinolytic response in the vascular wall and can play an important role in limiting thrombus growth. The incidence of thrombotic disease increases with age. As many adrenergic functions are affected by aging, the vascular response to adrenergic stimulation was studied in young (3 +/- 1 months) and aged (18 +/- 3 months) rats, using an experimental model of vascular perfusion. Five min epinephrine (EPI) infusion (0.15-25 microM) induced a dose-dependent increase in tissue-type plasminogen activator (t-PA) release and in perfusion pressure, in both young and aged rats. In the latter, however, the basal activity of t-PA was significantly higher than in young animals (0.22 +/- 0.02 vs 0.13 +/- 0.01 U/ml; means +/- SE; p < 0.01, n = 40) and EPI infusion induced a significantly higher increase in t-PA activity (0.78 +/- 0.06 vs 0.47 +/- 0.08 U/ml, p < 0.01, for 6.25 microM EPI). The response was characterized by a delayed onset. In contrast, the increase in the perfusion pressure by EPI was significantly lower in aged that in young rats, its basal levels being normal. To evaluate whether the increased vascular response of t-PA was peculiar for adrenergic stimulation, we infused platelet activating factor (PAF), an inducer of t-PA release, not related to catecholamines. PAF, during five-minute infusion induced a rapid and dose-dependent (100-1250 pM) increase in t-PA release, to the same extent in both young and aged rats. In conclusion, basal t-PA secretion was increased in aged rats probably as a response of vascular endothelium to chronic stimulation.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7482414 TI - Differential inhibition with antifibrinolytic agents of staphylokinase and streptokinase induced clot lysis. AB - The inhibitory effects of antifibrinolytic amino acids on clot lysis induced with recombinant staphylokinase (SakSTAR) or with streptokinase (SK) were evaluated in a human plasma milieu in vitro and in a hamster pulmonary embolism model in vivo. Addition of tranexamic acid to a system composed of 60 microliters 125I-fibrin labeled plasma clots submerged in 0.5 ml human plasma, caused dose-dependent inhibition of lysis; complete lysis in 120 min required 30 nM SakSTAR or 100 nM SK and was reduced to 50% with 0.015 mM or with 0.07 mM tranexamic acid, respectively. Aprotinin also produced dose-dependent inhibition; lysis with SakSTAR or with SK was reduced to 50% of the control value with 8 KIU/ml or with 10 KIU/ml aprotinin, respectively. Thus, in human plasma in vitro the antifibrinolytic potency of tranexamic acid was 5-fold higher towards SakSTAR than towards SK, whereas that of aprotinin was comparable towards both agents. In hamsters with pulmonary embolism given 0.063 mg/kg SakSTAR or 0.20 mg/kg SK over 30 min, the antifibrinolytic potency of tranexamic acid, administered as a single bolus injection or as a bolus injection followed by continuous infusion, was 8- to 10-fold higher towards SakSTAR than toward SK (50% reduction of clot lysis with SakSTAR at 12.5 mg/kg, as compared to 100-150 mg/kg with SK). In contrast, aprotinin was equipotent towards SakSTAR and SK (50% reduction of clot lysis with 2,000 to 2,700 KIU/kg).(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7482415 TI - Platelet accumulation on fibrin-coated polyethylene: role of platelet activation and factor XIII. AB - Platelet accumulation on small- and medium-calibre vascular grafts plays a significant role in graft occlusion. We examined platelet accumulation on the surface of fibrin-coated polyethylene tubing (internal diameter 0.17 cm) during 10 min flow (10 ml/min) at high wall shear rate (764 s-1). Washed platelets labelled with 51Cr were resuspended in Tyrode solution containing albumin, apyrase and red blood cells (hematocrit 40%). When the thrombin that was used to form the fibrin-coated surface was inactivated with FPRCH2Cl before perfusion of the tubes with the platelet: red blood cell suspension, the accumulation of platelets was 59,840 +/- 27,960 platelets per mm2, whereas accumulation on fibrin with residual active thrombin was 316,750 +/- 32,560 platelets per mm2 (n = 4). When the fibrin on the surface was cross-linked by including recombinant factor XIII (rFXIII) in the fibrinogen solution used to prepare the fibrin-coated surface, platelet accumulation, after thrombin neutralization, was reduced by the cross-linking from 46,974 +/- 9702 to 36,818 +/- 7964 platelets per mm2 (n = 12, p < 0.01). Platelet accumulation on tubes coated with D-dimer was ten times less than on tubes coated with D-domain; this finding also supports the observation that cross-linking of fibrin with the formation gamma-gamma dimers reduces platelet accumulation on the fibrin-coated surface. Thrombin-activated platelets themselves were shown to cross-link fibrin when they had adhered to it during perfusion, or in a static system in which thrombin was used to form clots from FXIII-free fibrinogen in the presence of platelets.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7482416 TI - KRDS, a peptide derived from human lactotransferrin, inhibits thrombin-induced thromboxane synthesis by a cyclooxygenase-independent mechanism. AB - KRDS, a tetrapeptide from human lactotransferrin, inhibits thrombin-induced platelet aggregation, secretion and thromboxane (TX) synthesis without interfering with phospholipase C (PLC) beta activation, since in previous work we have shown that Ca2+ mobilization and phosphorylation of the myosin light chain kinase (20 kDa) and pleckstrin (47 kDa) were normal. However, the inhibition of arachidonic acid-induced aggregation in the presence of KRDS is accompanied by normal TX synthesis suggesting that it does not interfere with the cyclooxygenase activity. To elucidate further the mechanisms of action of this peptide we tested its effect on U46619-induced platelet activation. KRDS inhibits U46619-induced platelet aggregation time- and dose-dependently without inhibiting the phosphorylation of pleckstrin. This suggests that the PLC pathway is not affected and that the inhibitory effect of KRDS is not due to and uncoupling of TXA2 from its receptor. In addition to the PLC pathway, protein tyrosine kinases play a major role in platelet signal transduction mechanisms. At least 7 tyrosine phosphorylated proteins are detected upon stimulation of platelets by thrombin. KRDS strongly inhibits the tyrosine-phosphorylated substrates, in particular two 100-105 kDa substrates which are related to GP IIb/IIIa activation and platelet aggregation. The absence of TX synthesis observed in the presence of KRDS could be due to the inactivation of cPLA2 since the latter needs tyrosine phosphorylation to be activated, thus explaining the inhibitory action of KRDS on platelet functions. PMID- 7482417 TI - Regulation of platelet aggregation by post-fibrinogen binding events. Insights provided by dithiothreitol-treated platelets. AB - A variety of data suggest that fibrinogen binding is necessary but not sufficient for platelet aggregation: post fibrinogen binding events may play an important role. The present study compared fibrinogen binding and platelet aggregation in response to dithiothreitol (DTT) and ADP. DTT induced saturable and specific fibrinogen binding (Kd 0.07 + 0.02 microM, Bmax 15,000 + 3000 molecules/platelet) which supported complete platelet aggregation as determined by single platelet counting. The aggregates were small, however, and more readily dissociated by EDTA than their ADP-treated counterparts, despite quantitatively similar fibrinogen binding. Unlike fibrinogen bound to ADP-stimulated platelets, fibrinogen bound to DTT-treated platelets remained sensitive to dissociation by EDTA over a 3 h time course, retained its ability to support aggregation, even when aggregation was induced 60 min after the initial platelet exposure to fibrinogen, and remained accessible to polyclonal antibodies and plasmin. Confocal scanning laser microscopy showed only a surface clustering of fibrinogen bound to DTT-treated platelets over the 3 h time course compared to rapid fibrinogen clearing from the surface of ADP-stimulated platelets. These data suggest that post fibrinogen binding events involved in the stabilization of fibrinogen binding and/or the redistribution of bound fibrinogen may play important roles in regulating platelet aggregation. PMID- 7482418 TI - Vessel wall injury and arterial thrombosis induced by a photochemical reaction. AB - Arterial thrombosis may be initiated in an experimental animal by a photochemical reaction between transmural green light and i.v. administered Rose Bengal, a photosensitizer dye. In this study, scanning electron microscopy has been used to reveal the nature of vessel injury and the cellular composition of the photochemically induced thrombus. A 5 mm segment of the guinea pig femoral artery was occluded by a thrombus about 10 min after irradiation with green light in the presence of systemically administered Rose Bengal. Electron microscopy revealed that following photochemical reaction, endothelial cells first contract and, with further irradiation, become detached from the vessel wall, with their cell membrane being destroyed at the irradiated site where an occlusive platelet-rich thrombus was formed. Endothelial cell injury and vessel occlusion could be completely inhibited by the aminothiol, DL-cysteine administered i.v. 1 min after Rose Bengal. The mechanism of endothelial injury in this model appears to be by singlet molecular oxygen, 1O2 formed by energy transfer from the photo-excited dye to O2. PMID- 7482419 TI - Sites of tissue factor pathway inhibitor (TFPI) and tissue factor expression under physiologic and pathologic conditions. On behalf of the Subcommittee on Tissue factor Pathway Inhibitor (TFPI) of the Scientific and Standardization Committee of the ISTH. AB - Under normal conditions, TFPI expression is restricted to megakaryocytes and the endothelium of the microvasculature. It is not synthesized by normal hepatocytes or by the endothelium of larger vessels. In contrast, endothelium and peripheral blood cells do not express tissue factor under normal conditions. Expression of tissue factor under normal physiologic conditions is widespread and is localized in areas which are physically separated from the circulating blood. During an inflammatory response, circulating monocytes have been shown to express tissue factor, whereas in general, expression of tissue factor has not been observed in the endothelium. Adherent monocytes/macrophages express both tissue factor and TFPI under pathologic conditions. Whether or not circulating peripheral blood monocytes under inflammatory conditions express TFPI is not known. PMID- 7482420 TI - Protein C deficiency: a database of mutations, 1995 update. On behalf of the Subcommittee on Plasma Coagulation Inhibitors of the Scientific and Standardization Committee of the ISTH. PMID- 7482421 TI - Which patients undergoing cardiopulmonary bypass should be assessed for development of heparin-induced thrombocytopenia? PMID- 7482422 TI - High-dose intravenous immunoglobulin delays clearance of von Willebrand factor in acquired von Willebrand disease. PMID- 7482423 TI - Dietary fat intake and plasma factor VII antigen concentration. PMID- 7482424 TI - The effect of ionizing radiation on endothelial tissue factor activity and its cellular localization. PMID- 7482425 TI - Molecular analysis in factor XIIIA deficiency. PMID- 7482426 TI - Catalyzing global access to hemophilia care. PMID- 7482427 TI - XVth Congress of the International Society on Thrombosis and Haemostasis. Jerusalem, Israel, June 11-16, 1995. Abstracts. PMID- 7482428 TI - Inhibition of platelet aggregation in whole blood by alcohol. AB - Our previous studies have demonstrated that addition of moderate volumes of absolute alcohol (34-170 mM final concentration) to whole blood produces concentration-dependent platelet aggregation, due to release of adenosine diphosphate (ADP) from erythrocytes. We have now investigated the effects of exposure of blood to ethanol by a more "physiologic" protocol, in which 7.8% (w/v) alcohol is added to achieve a final concentration of 1 to 85 mM in human and rat blood or platelet rich plasma (PRP). The effects of short incubation with alcohol on platelet aggregation induced by ADP, collagen and arachidonic acid were examined by the impedance method of aggregometry. Aggregation induced by collagen in PRP of either species was significantly inhibited by 85 mM ethanol, with concentrations as low as 4.25 mM inhibiting the response to collagen in rat whole blood. ADP stimulated only primary, reversible aggregation in rat PRP and whole blood, and these responses were unaffected by alcohol. Human platelets responded to ADP with irreversible aggregation, which was significantly attenuated by 85 mM ethanol in whole blood but not PRP. Arachidonic acid evoked irreversible platelet aggregation in all four preparations; this was significantly inhibited by the high dose ethanol in human and rat PRP, but not whole blood. In contrast to our earlier studies with absolute ethanol, there was no evidence of hemolysis (and therefore, ADP release from red blood cells) using the current protocol. The results of these experiments show that alcohol, at physiologically relevant concentrations, has an inhibitory effect on secondary platelet aggregation responses to some agonists in whole blood as well as PRP, possibly by its previously demonstrated effects on arachidonic acid release by phospholipases. The possibility remains to be considered that other blood cells might contribute to the effects of alcohol on platelet aggregation in whole blood. PMID- 7482429 TI - Endotoxin-induced pulmonary vascular injury is mainly mediated by activated neutrophils in rats. AB - Acute respiratory failure is a common complication in patients with disseminated intravascular coagulation associated with sepsis. To elucidate the role of coagulation abnormalities in acute lung injury in sepsis, we investigated the effect of anticoagulants on the pulmonary vascular injury in rat induced by lipopolysaccharide (LPS). When administered intravenously, LPS (5 mg/kg body weight) significantly increased the accumulation of 111indium-labeled neutrophils in lung 30 min after administration. Subsequently, the pulmonary vascular permeability and the serum level of fibrin and fibrinogen degradation products (E) [FDP (E)] increased and remained elevated for several hours. Neither heparin alone, heparin plus antithrombin III, or dansyl-Glu-Gly-Arg-chloromethyl ketone treated factor Xa, a selective inhibitor of thrombin generation, prevented LPS induced vascular injury 6 hours after LPS administration, whereas these substances significantly inhibited the increase in serum FDP (E) at that time. LPS-induced pulmonary vascular injury was significantly attenuated in rats with methotrexate-induced leukocytopenia or treated with ONO-5046, a potent granulocyte elastase inhibitor, although ONO-5046 did not inhibit the LPS-induced increase in serum FDP (E). Thus, activated leukocytes play a more important role than coagulation abnormalities in the pathogenesis of LPS-induced pulmonary vascular injury in an experimental rat model of endotoxemia. PMID- 7482430 TI - Antithrombin III deficiency as a risk factor for catheter-related central vein thrombosis in cancer patients. AB - The fibrin sleeve of venous catheters (VC) and parietal thrombi represent frequent and dangerous side-effects of central venous catheterization (CVC), due to the risk of embolism. Reduced levels of coagulation clotting factors inhibitors (such as Antithrombin III) are known to be associated with increased thrombogenic risk. The aim of this study was to evaluate the role of Antithrombin III (AT III) deficiency as a risk factor for thrombosis in cancer patients undergoing CVC. The study groups included patients with a reduced AT III activity (< 70%, 20 consecutive patients) and with normal AT III values (> 70%, 20 randomly selected patients), requiring a VC for chemotherapy and/or total parenteral nutrition. The study protocol included evaluation of Hb, PLTs, PT (INR), aPTT, Fibrinogen and AT III at days 0, 1, 3 and 8 after CVC and upon VC removal. Peripheral and pullout phlebographies were performed in all patients on catheter withdrawal. A quantitative scale was developed to evaluate both VC and parietal thrombus degree in each catheter-containing venous segment (subclavian, innominate, superior vena cava); the sum of the mean values was defined as overall thrombus. The average VC dwelling time was similar in both groups. There were no significant differences in Hb, PLTs, PT (INR), aPTT, Fibrinogen and in the remaining parameters of the study between the two groups. The group with AT III deficiency presented a higher degree of both parietal (p < 0.05) and overall thrombus (p < 0.02). Data showed a higher severity of CVC-related thrombosis in patients with AT III deficiency than in the control group. Further studies are needed to evaluate whether the therapeutically-induced normalization of AT III levels can reduce the thrombosis degree. PMID- 7482431 TI - The injection of heparin prolongs the plasma clearance of oxidized low density lipoprotein in the rat. AB - There is evidence that oxidized-LDL plays an important role in atherogenesis. We now report on the in vivo interaction between unfractionated heparin and oxidized LDL in rats. The recovery rates of the native LDL particles ranged between 75% and 85% of the injected dose. Heparin did not interfere with the clearance rates of native LDL. After administration of radioactive labeled oxidized-LDL particles, 26% of the material was measured in circulation after 5 minutes, 8% after 20 minutes, and 3% after 60 minutes. After injection of heparin 2 minutes prior to oxidized-LDL tracer particles, 44% of the tracer was found in blood after 5 minutes, 23% after 20 minutes, and 9% after 60 minutes. Oxidized-LDL tracer particles disappeared from blood with an alpha half-life of 5 minutes and a beta half-life of 7.5 minutes. After receptor blocking with unfractionated heparin the alpha half-life of the oxidized-LDL tracer was prolonged to 17.5 minutes and the beta half-life to 27.5 minutes. These results indicate that heparin molecules of a comparatively small molecular weight competed the scavenger receptor mediated uptake of oxidized-LDL particles in vivo. Oxidized LDL particles are known to mediate their pro-atherosclerotic activity in part by stimulating smooth muscle cell proliferation by a scavenger receptor-mediated pathway. It can be speculated, if heparins interfere with the uptake of oxidized LDL, heparins might thus in part exert their known antiatherosclerotic properties. PMID- 7482433 TI - Remnant-like lipoproteins stimulate whole blood platelet aggregation in vitro. AB - We have developed a simple, rapid assay method to measure remnant-like lipoproteins by using an immunoaffinity gel mixture of anti apo B-100 and apoA-1 antibodies to Sepharose 4B. Characterization of the unbound lipoproteins has shown that they represent chylomicron and VLDL remnant particles (RLP). Preincubation of whole blood with RLP resulted in the enhanced activation of aggregation with ADP and collagen. Such enhancement was not observed in the presence of lipoprotein deficient serum or albumin preparation. The extent of enhancement was 2.78 times by 7.5 microM of ADP and 44 times by 0.5 microgram/ml of collagen in the presence of RLP-preparation 1 (RLP-1), respectively. In the presence of RLP-2, the enhancement was 5.37 times by 7.5 microM of ADP and 102 times by 0.5 microgram/ml of collagen, respectively. On the other hand RLP slightly inhibited PRP aggregation by these agonists. Inhibitions were 19% by 7.5 microM of ADP and 18% by 1.0 microgram/of collagen in the presence of RLP-1, respectively. Incubation of whole blood with RLP did not result in the release of factors to stimulate platelets or ADP- or collagen-induced platelet aggregation in vitro. The extents of enhanced aggregation in whole blood or inhibition in PRP were not correlated with RLP-cholesterol nor RLP-protein concentrations of RLP preparations used. These results may indicate that RLP not only interact with platelets but with erythrocytes or leukocytes. Our findings support the hypothesis that the postprandial increase in remnant lipoproteins is an atherosclerotic risk factor and may be a part of the reasons of thrombotic complications by stimulating platelets in patients with remnant hyperlipoproteinemia. PMID- 7482432 TI - Inhibition of platelet aggregation and eicosanoid production by phenolic components of olive oil. AB - This study was designed to investigate the in vitro effects of phenolic compounds extracted from olive oil and from olive derived fractions. More specifically, we investigated the effects on platelets of 2-(3,4-di-hydroxyphenyl)-ethanol (DHPE), a phenol component of extra-virgin olive oil with potent antioxidant properties. The following variables were studied: aggregation of platelet rich plasma (PRP) induced by ADP or collagen, and thromboxane B2 production by collagen or thrombin stimulated PRP. In addition, thromboxane B2 and 12-hydroxyeicosatetraenoic acid (12-HETE) produced during blood clotting were measured in serum. Preincubation of PRP with DHPE for at least 10 min resulted in maximal inhibition of the various measured variables. The IC50s (concentration resulting in 50% inhibition) of DHPE for ADP or collagen-induced PRP aggregations were 23 and 67 microM, respectively. At 400 microM DHPE, a concentration which completely inhibited collagen-induced PRP aggregation, TxB2 production by collagen- or thrombin-stimulated PRP was inhibited by over 80 percent. At the same DHPE concentration, the accumulation of TxB2 and 12-HETE in serum was reduced by over 90 and 50 percent, respectively. We also tested the effects of PRP aggregation of oleuropein, another typical olive oil phenol, and of selected flavnoids (luteolin, apigenin, quercetin) and found them to be much less active. On the other hand a partially characterized phenol enriched extract obtained from aqueous waste from olive oil showed rather potent activities. Our results are the first evidence that components of the phenolic fraction of olive oil can inhibit platelet function and eicosanoid formation in vitro, and that other, partially characterized, olive derivatives share these biological activities. PMID- 7482434 TI - Mechanism of inhibition of fibrinolysis and fibrinogenolysis by the end fibrinogen degradation products. AB - Influence of the end fibrinogen-derived DH, DL and E fragments on fibrinolysis and fibrinogenolysis has been studied. Electrophoresis of the plasmin-digested unstabilized fibrin and fibrinogen showed that fragment E was the only inhibitor of plasmin hydrolysis of fibrinogen, the antifibrinolytic activity of the fragments being increased in a DL < E < DH series. The fragments were revealed by means of elastic light scattering and analytical ultracentrifugation to be arranged in a E > DL > DH series by their ability to form a complex with plasminogen. It was concluded that the complex formation did not greatly contribute to the mechanism of fibrinolysis inhibition. Antifibrinolytic effect of fragment DH is due to its antipolymerization activity. The paper discusses the competitive protein-protein interactions occurring on a polymeric matrix of fibrin. PMID- 7482435 TI - False-positive activated protein C (APC) sensitivity ratio caused by freezing and by contamination of plasma with platelets. PMID- 7482436 TI - Fibrinogen Guarenas I: partial characterization of a new dysfibrinogenemia with an altered rate of fibrinopeptide release and an impaired polymerization. AB - A congenitally abnormal fibrinogen was isolated from the blood of a young woman with a severe bleeding diathesis. Coagulation tests showed a prolonged Thrombin and Reptilase time partially corrected by Ca2+. Polymerization of thrombin induced preformed fibrin monomers was severely impaired. Thrombin caused the release of fibrinopeptides with normal retention times on HPLC. However, the rate of release was abnormally slow and the total amount of fibrinopeptide A (FpA) released reached only approximately 50% of the theoretical maximum. The rate and quantity of FpA release was normal when Reptilase was used. Transmission Electron Microscopy (TEM) of Thrombin induced clots showed an altered clot structure characterized by a reduced mean fiber diameter. The mother has a polymerization defect similar to the propositus, her fibrinopeptide release is unaffected however. The father has a minor fibrinopeptide release defect suggesting the presence of two populations of fibrinogen. This study supports the idea that the fibrinogen isolated from the propositus has two defects inherited as separate genetic traits. This fibrinogen has been named Fibrinogen Guarenas I. PMID- 7482437 TI - Effect of in vitro incorporation of prostanoid precursors, superoxide radical and hydrogen peroxide on platelet function. AB - The level of cyclic adenosine monophosphate (cAMP) in human platelets is known to be an important regulator of platelet function. The polyunsaturated fatty acids (PUFA) dihomo-gamma-linolenic acid (DHLA), and eicosapentaenoic acid (EPA), precursors of the prostaglandin (PG) 1 and 3 series respectively, were studied for their ability to stimulate platelet cAMP and/or PGE1 levels, and to inhibit platelet aggregation (PAg). Incubation of washed platelets (1 x 10(8)/ml) with 125 microM DHLA increased intraplatelet levels of PGE1 from 197 +/- 7 to 1622 +/- 9.7 picograms/10(8), cAMP from 3 +/- 0.8 to 31 +/- 1.9 picomoles/10(8), and inhibited collagen-induced PAg. Addition of 1 mumole of xanthine per unit of xanthine oxidase (a superoxide radical generating system) to the incubating medium potentiated the effects of both fatty acids, whereas 240 microM Hydrogen Peroxide (H2O2) inhibited these effects. These results suggest that: (1) DHLA may be more effective in inhibiting PAg than EPA, which has been reported to reduce the incidence of coronary diseases in some human populations; (2) That superoxide radical may activate the platelet cyclooxygenase system to increase lipid peroxidation of these PUFA prostanoid precursors and may result in the inhibition of PAg, whereas H2O2 may have an opposite effect. PMID- 7482438 TI - Urokinase binding and catabolism by Hep G2 cells is plasminogen activator inhibitor-1 dependent, analogous to interactions of tissue-type plasminogen activator with these cells. AB - The adherent human hepatoma cell line Hep G2 exhibits receptor mediated endocytosis and catabolism of tissue-type plasminogen activator.plasminogen activator inhibitor type-1 (t-PA.PAI-1) complexes formed when exogenous t-PA combines with endogenous PAI-1 in the extracellular matrix. To determine whether the other major PA, urokinase (u-PA), which also complexes with PAI-1, is metabolised via the same mechanism, 125I-labelled high (hmw) and low (lmw) molecular weight forms of u-PA were incubated with Hep G2 cells at 4 degrees C for 2 hr in the absence and presence of a 100-fold excess of unlabelled ligand in order to detect specific binding. Both hmw and lmw 125I-u-PA formed complexes with PAI-1 and these bound specifically and with high affinity (apparent Kd 3.9 and 4.1 nM, with Bmax 78 x 10(3) and 83 x 10(3) binding sites/cell respectively). Binding by each form of radiolabelled u-PA was inhibited in a dose-dependent fashion by unlabelled t-PA, hmw-u-PA, lmw-u-PA, and by monoclonal anti-PAI-1 antibody. At 37 degrees C, bound hmw and lmw 125I-u-PA.PAI-1 complexes were internalised and degraded rapidly. These findings indicate that the specificity of the previously described receptor which mediates PAI-1 dependent catabolism of t-PA by Hep G2 cells extends to complexes of u-PA with this inhibitor. PMID- 7482439 TI - Decreased fibrinolytic stimulation by a short-term venous occlusion test in patients with cerebrovascular disease. AB - The aim of the present study was to determine whether patients with previous cerebrovascular disease have abnormalities in hemostatic functions stimulated by a 5-min venous-occlusion-test. Twenty-two patients with brain ischaemia 3-6 months previously were compared to a control group of twenty patients with non vascular neurological diseases. Cases showed less increase in tissue plasminogen activator and fibrin degradation products than controls. There were no differences in baseline and stimulated levels of plasminogen activator inhibitor, fibrinogen degradation products, thrombin-antithrombin complex or global lysis assay (Fibriscreen). Since patients with acute cerebrovascular disease were not included, the present findings may represent either preexisting or longterm reactive fibrinolytic deficiencies, possibly predisposing to further thrombotic disease. PMID- 7482440 TI - Reductions in tissue plasminogen activator and thrombomodulin in blood draining veins damaged by venous access devices. AB - A frequent complication of venous access devices (VADs) is axillary-subclavian venous thrombosis. To study this problem we have compared blood drawn through VADs with peripheral blood samples in a group of oncology patients with venographically demonstrated venous damage (N = 14) and a group with normal venograms (N = 21). The samples were assayed for a battery of proteins believed to be involved in thrombogenesis. After approximately six weeks of catheterization the venographically abnormal patients had significantly less thrombomodulin (P = 0.0055) and significantly higher PAI:tPA (P = 0.022) in catheter-drawn samples as compared with the venographically normal group. Although the data are inconclusive, it is hypothesized that these changes resulted from local endothelial injury. PMID- 7482441 TI - Evidence against hypercoagulability in coronary artery disease. AB - The strong epidemiological association between elevated plasma clotting factors and coronary artery disease is generally interpreted as evidence that patients with coronary atherosclerosis are in a procoagulant (hypercoagulable) state. A dynamic global test was used to assess the overall coagulation status of 761 patients with coronary artery disease scheduled for coronary artery bypass grafting and compared to healthy matched controls (n = 100). Platelet reactivity to shear-stress was simultaneously measured from identical, non-anticoagulated blood samples. Contrary to expectation, the overall coagulation in cardiac patients did not differ significantly from that of controls. Furthermore, the coagulation status of patients bore no relationship to the severity of coronary atherosclerosis. The latter is in contrast with platelet reactivities, which were significantly increased in patients with > or = 2 vessel disease as compared with single vessel disease. The present results do not necessarily conflict with the finding of elevated plasma clotting factors in cardiac patients. However, they do not support the claim that these markers are a reflection of a hypercoagulable state. Indeed, this study confirms that such patients are in a prothrombotic state, which is related to enhanced platelet reactivities, and not to a prothrombotic imbalance of the coagulation mechanism. PMID- 7482442 TI - Thrombin induces GRO alpha/MGSA production in human umbilical vein endothelial cells. AB - Thrombin, besides being a potent coagulation factor, exerts influence on endothelial and leukocyte functions and may thus be involved in the regulation of inflammatory reactions. The present study investigated whether thrombin stimulates the production of growth-related cytokine/melanoma growth-stimulatory activity (GRO alpha/MGSA) in endothelial cells. Human umbilical vein endothelial cells (HUVEC) stimulated with thrombin were found to product GRO alpha/MGSA in a dose- and time-dependent manner. This action of thrombin was completely suppressed by preincubation with either hirudin or antithrombin-III (AT-III) heparin. Interestingly, the thrombin receptor-activating peptide SFLLRN mimicked the action of thrombin. In addition, staurosporine, a protein kinase C (PKC) inhibitor, attenuated the production of GRO alpha/MGSA by thrombin, SFLLRN and phorbol 12-myristate 13-acetate (PMA), but left the action of interleukin-1 beta (IL-1 beta) unchanged. These results suggest that catalytic activation of thrombin receptor by thrombin results in GRO alpha/MGSA production, at least in part, via a pathway involving PKC in HUVEC. PMID- 7482443 TI - Functional assay of plasma antithrombin using polyethylene glycol (PEG) defibrinated plasma. AB - Polyethylene glycol(PEG) was used to precipitate fibrinogen to prepare defibrinated plasma in the two stage clotting assay of antithrombin activity. Five percent PEG-8000 precipitated fibrinogen from plasma without loss of antithrombin activity in the defibrinated plasma. Fibrin degradation products(FDP) as high as 640 ug/ml did not interfere the two stage clotting assay using PEG defibrinated plasma possibly because part of FDP was precipitated by PEG in the process of plasma defibrination. The two stage clotting assay was very sensitive to the changes of antithrombin activity in the range of 60%-100% of normal level. The assay was reproducible and correlated with chromogenic assay. The decrease of plasma antithrombin activity in a baboon septic shock model was demonstrated with this assay. PMID- 7482444 TI - Characterisation of the chains of human fibrinogen isolated by perfusion chromatography using fibrin specific monoclonal antibodies. AB - In order to study the epitopes on fibrin to which monoclonal antibodies are directed, we required pure individual polypeptide chains of human fibrinogen in milligram quantities. High purity chains of human fibrinogen were rapidly obtained, in under 3 minutes, by the novel procedure of reversed-phase perfusion chromatography and these chains were subjected to immunological characterisation using monoclonal antibodies specific to the individual chains. Cross-reactivity against these antibodies in both immunoblotting and enzyme linked immunospecific assay (ELISA) procedures showed that these isolated fibrinogen chains were of high purity and retained high immunoreactivity. These chains were employed to initiate studies to define the epitopes in fibrin to which four fibrin specific monoclonal antibodies, B10, A11, 5F3 and 1H10 are targeted. Two of these antibodies, B10 and A11, were shown to be directed to a linear sequence on the A alpha chain, although the binding profiles for the two antibodies suggested that different epitopes may be involved for each of these two antibodies. MAbs, 1H10 and 5F3, however, did not bind to any of the three fibrinogen chains, suggesting that conformational epitopes in fibrin are likely to be involved in the binding of these two antibodies to fibrin. PMID- 7482445 TI - Effect of TNF-alpha, Shiga toxin and calcium ionophore on Weibel-Palade body content of endothelial cells: possible implications for the hemolytic uremic syndrome. PMID- 7482446 TI - Activating effect of the plasminogen activators on plasminogens of different mammalia species. PMID- 7482447 TI - [Early diagnosis of colorectal tumors]. PMID- 7482448 TI - [MUTAN--an AIDS project in Tanzania]. PMID- 7482449 TI - [Quality assurance in health care--a paper tiger?]. PMID- 7482450 TI - [Malignant non-Hodgkin's lymphomas. Classification, treatment and survival in a 10-year material]. AB - Over a ten year period, 71 cases of malignant non-Hodgkin's lymphomas were treated at the Department of Internal Medicine, Nordland Central Hospital. Of these cases, 41 were low grade malignant non-Hodgkin's lymphomas, while 28 were high grade. 41% presented localized disease. 30% had primary extranodal manifestations. Median age was 68 years. 5-years disease-specific survival was 57%. The results of treatment are judged to be satisfactory when compared with the results from other unselected materials. Improvements can be made on some points. The study revealed possibly too extensive use of surgery in patients with extranodal manifestations, and too extensive use of aggressive chemotherapy in patients with low grade malignancy. PMID- 7482451 TI - [Diagnosis and treatment of lymphomas. Retrospective quality assessment of a 10 year material]. AB - We have performed a quality assessment of staging and treatment of 64 patients with non-Hodgkin's lymphoma treated at the Department of Internal Medicine, Nordland Central Hospital from 1982 to 1991. The assessment was based on defined quality criteria. Journal records of patient history, physical examination and stage were unsatisfactory. Histological examinations, use of laboratory tests and X-ray examinations were appropriate. The choice of chemotherapy for high grade malignancy was adequate, average relative dose-intensity was low. Low utilization of radiotherapy could be explained in most cases by individual patient factors. 5 years disease-specific survival was 54% for all patients and 70% for those presenting with localized disease. PMID- 7482452 TI - [Cholesterol determination at conscription. A method to identify persons with high risk of developing cardiovascular disease]. AB - In the adult population, serum cholesterol level and risk of cardiovascular disease are related to some extent to habits and lifestyle established at an early age. We have estimated serum total cholesterol levels by means of a dry chemical method and have collected information on established cardiovascular risk factors among 1,203 young Norwegian men at conscription. 30 of the recruits with the highest serum cholesterol levels were later examined in the hospital's out patient clinic. A total of 30.8% of the recruits were daily smokers. Mean serum total cholesterol was 4.05 mmol/l with a 97.5 percentile value of 6.31 mmol/l. The prevalence of coronary heart disease among parents was significantly higher among recruits from the upper cholesterol quintile (4.2%) compared with those in the lowest quintile (0.8%) (p = 0.02). These findings show that cholesterol screening at conscription is feasible and can be used to identify a group of men at high risk of subsequently developing cardiovascular disease. PMID- 7482453 TI - [Pathogenesis of infections--a complex interplay between microbes and the immune defence]. AB - The pathogenesis of infectious diseases is regulated by a complex interplay between microbes and the immune system. To function well, the immune system is dependent on factors in both the internal and the external environment. Behavioural, social and cultural factors are all important. Increasing resistance to drugs and greater awareness of other side effects of antibiotics have led to greater recognition by doctors of the need to use alternative ways of treating and preventing infections, particularly by modulating the immune response. Important elements of the defence against various types of infectious agents are outlined. This is intended as a framework for understanding how infectious diseases can be dealt with by modulating the immune response instead of only attempting to kill the microbe. This is particularly important in patients with serious infections and in groups of patients specially predisposed to infections. PMID- 7482454 TI - [New immunopathogenetic aspects of infectious diseases]. AB - The clinical course of infectious diseases depends on the complicated interaction between the microorganism and the immune system of the host. The vast amount of knowledge acquired during recent years in the field of immunology will probably lead to new ways of treating infectious diseases by selective immunomodulation. In this paper, we review some newer aspects of host defence mechanisms of relevance to immunomodulation of infectious diseases. The cytokine network, apoptosis, superantigens, and oxidative stress are all discussed with reference to various infectious diseases. Furthermore, the possible role of physical and psychological stress in the immunopathogenesis of infectious disease is discussed in the light of the interplay between the immune system and the neuroendocrine system. PMID- 7482455 TI - [Virus as an etiological factor of cancer in human beings]. AB - Human oncogenic viruses are heterogenous as a group but share the property of causing persistent and often lifelong infections. Immortalization of host cells is seen in some but not all infections. Chromosomal instability of the infected cells is probably important. Epidemiological data indicate that viral oncogenesis in humans is a multifactorial process where host factors such as age, race, sex and immune-competence, and exogenous factors such as coinfections and cocarcinogens, are important determinants. PMID- 7482456 TI - [Determination of the activated proto-oncogene (Ki-ras) in feces. A new laboratory analysis for early diagnosis of colorectal cancer]. AB - Colorectal cancer is one of the most common malignancies in Norway. Many cases of the disease are detected at a stage where surgery is unlikely to result in cure. Currently used screening tests based on faecal occult blood and other tumour markers are poor indicators of colorectal neoplasia. Multiple gene alterations are associated with colorectal carcinogenesis. Mutations in the Ki-ras oncogene occur in 50% of colorectal carcinomas and also in 50% of the precursor lesions, the adenomas. These mutations have proved to be detectable in the faeces of patients with colorectal tumours, but the techniques used so far have been impractical for screening purposes. We have developed a rapid and simple laboratory technique, based on the polymerase chain reaction, for detecting mutated Ki-ras in the faeces. In eight of 12 patients with mutated Ki-ras in adenomas or carcinomas we found the corresponding mutations in stool samples. Our results represent a significant progression towards a simple and effective DNA based screening strategy for early detection of curable colorectal cancer. PMID- 7482457 TI - [Pelvic floor relaxation--a condition with many names and uncertain criteria]. AB - Pain in the pelvis and lower back appears to be becoming an increasing problem in Norway. The relation between pelvic pain and low back pain is unclear. In the literature a variety of designations, such as pelvic girdle relaxation, pelvic joint instability, posterior pelvic pain, peripartum pelvic pain etc. have been used as identifying labels. The condition(s?) seems to be culturally specific for the Nordic countries in particular. However, pelvic pain is also recognized in England, Holland and Australia, but seemingly unknown in France, Germany, Italy and Spain. The article describes the current knowledge--focusing criteria, etiology, prevalence and nomenclature--and points to the need for systematic epidemiological and clinical investigations. PMID- 7482458 TI - [Storage mite allergy in Western Norway]. AB - We describe two different studies of storage mite allergy from the Stavanger area performed among 2,273 patients who were referred to ear, nose and throat specialists. Storage mite allergy is common in the western part of Norway. It is a major factor in the development of allergic rhinoconjunctivitis and asthma in rural areas and among allergic patients living in damp houses. We conclude that, in western Norway, storage mites should be included in standard skin test when allergic rhinoconjunctivitis and/or asthma is suspected. PMID- 7482459 TI - [The HIV-epidemic in North-East Tanzania]. AB - A series of seven population-based HIV-screening surveys were conducted in Arusha region between November 1991 and April 1994. Communities included three wards within Arusha town, two townships along major highways in Arusha region, and two rural villages. A total of 3,440 people completed an interview to assess AIDS related knowledge, attitudes and practices, and of these, 1,932 consented to be tested for HIV (56.2%). Across communities, 7.4% of the participating women and 1.9% of the men tested HIV-positive. For both sexes the HIV-prevalence was significantly higher in urban than in rural areas. As many as 12.5% among young women from Arusha town were HIV-positive. HIV-prevalence was higher among participants from urban areas characterized by lower socio-economic status than among participants living in areas of higher socio-economic status. These surveys tend to confirm that women and poor people are the most vulnerable to HIV infection. The local HIV-epidemic seems to spread from urban to rural areas along major highways. PMID- 7482460 TI - [AIDS preventive work in Arusha and Kilimanjaro--health education]. AB - Over the past five years the Tanzanian-Norwegian AIDS Project (MUTAN) has assisted the National AIDS Control Programme in creating and testing innovative HIV/AIDS educational programmes. These programmes, designed to reach a variety of target groups, have been implemented throughout the Arusha and Kilimanjaro regions and include: public meetings, intensive courses, information centres, radio programmes and school-based programmes. A recent survey of 996 participants (15-54 years old) was designed to assess public exposure to HIV/AIDS information. A total of 72% of the participants reported having heard about AIDS on the radio, 74% having read about AIDS in newspapers, and 52% having heard about AIDS from a health worker during the previous month. Furthermore, 26% had listened to MUTAN's weekly radio programme at least once. 31% knew of MUTAN's information centres, and 15% had visited one of these centres. The results indicate that large proportion of the population is receiving in-depth HIV/AIDS information. It is recommended that future work concentrate on how to reach people with no or little formal education, young adults and women. PMID- 7482461 TI - [Bar attendants as health educators. Prevention of sexually transmitted diseases in high-risk areas]. AB - In Tanzania, a nation-wide programme against the spread of HIV is targeting villages along the main highways, mining and construction centres, and border towns. These places contain a high number of bars and guest houses, where female bar attendants offer sexual favours to obtain a minimum income for survival. The strategy is to elect and educate bar workers as "peer health educators", motivate them to reduce the number of sexual partners, to distribute and promote use of condoms, and to establish easier access to treatment for sexually transmitted diseases. The activities are monitored and supervised through regular visits by trained health behaviour officers. The Tanzanian-Norwegian AIDS project has implemented the programme in Arusha and Kilimanjaro regions. PMID- 7482462 TI - [Health care seeking behavior of patients with AIDS]. AB - We describe an anthropological study of the disease histories of ten persons with AIDS in Tanzania. It focuses on the health care-seeking behaviour of these patients. AIDS has many clinical manifestations, and it is for these manifestations that the AIDS patient seeks treatment. The case studies suggest that people with AIDS seek many different forms of treatment, both biomedical and "traditional", and will travel far and wide to obtain the best possible treatment. People with AIDS are usually discharged from the hospitals. The governmental health care service has little to offer them in terms of care and medicine. A majority of the AIDS sick are cared for at home by their own families. Home-based care is an important factor in the management of the disease, but has a number of constraints. PMID- 7482463 TI - [Counseling and HIV-testing among hospital patients in Arusha and Kilimanjaro]. AB - In 1989 the Ministry of Health in Tanzania decided that all patients suspected to be HIV-infected should be counselled in order to prevent or reduce the risk of HIV-transmission and to help people cope with the disease. This article describes the introduction of a counselling service at the Regional Hospitals of Arusha and Kilimanjaro and provides an overview of our experiences during the period 1990 92. Patients at the two hospitals received pre-test, post-test and follow-up counselling. Of 1,194 persons pre-test counselled, 95% gave their informed consent to be tested 62% were HIV-positive. Data on clinical pretest assessment of the patients matched the HIV-test assessment of the patients matched the HIV test results for those assessed to have full blown AIDS, but there was more discrepancy for patients assessed to have early signs of AIDS or to be in the terminal phase. The majority of the HIV-positive patients accepted their diagnosis when informed. The routines at the hospitals are still functional five years after the introduction of the counselling service. PMID- 7482464 TI - [Introduction and evaluation of HIV tests in the North-East part of Tanzania]. AB - The MUTAN (Tanzanian Norwegian AIDS Project) virology programme has comprised research, intervention, surveillance and education as part of the Tanzanian National AIDS Control Programme. HIV testing of blood donors was introduced in 1988 in the regions Arusha and Kilimanjaro. Simple and rapid HIV-tests have been evaluated continuously, as well as the possible use of alternative specimen samples for testing. The polymerase chain reaction for detection of HIV proviral DNA was established at the Northern Zone Reference Hospital, The Kilimanjaro Christian Medical Centre in Moshi, in 1993. PMID- 7482465 TI - [On solved mysteries and false expectations]. PMID- 7482466 TI - [Q10 and heart failure]. PMID- 7482467 TI - [Development of computer-assisted learning programs and encyclopedias]. PMID- 7482468 TI - [Evaluation of a course on instruction in medical education for hospital specialitst]. PMID- 7482469 TI - [When ethics interfere with research]. PMID- 7482470 TI - [The abnormal white line and hollow wall in horses, a comparative mycological study]. AB - There are few descriptions of an abnormal white line in the horse, and it is not known what causes this disorder. Thirty horses and ponies with this disorder were examined mycologically, as were 30 horses and ponies with normal hooves. Scopulariopsis brevicaulis was detected in 60% of the animals with an abnormal white line with or without hollow wall or brittle toe. PMID- 7482471 TI - [A study of the cause of massive mortality among marine-cultured rainbow trout (Oncorrhynchus mykiss, Walbaum) in a fish farm in the southwestern Netherlands]. AB - The present study investigated the mass mortality of marine cultured young rainbow trout (Oncorhynchus mykiss, Walbaum) in the Netherlands. The course of the disease, the clinical symptoms, and bacteriological and virological investigations lead to the diagnosis: 'primary infectious pancreatic necrosis virus (IPNV) infection followed by secondary vibriosis'. Treatment with flumequine seemed to be effective. The trout were possible infected with IPNV at the trout hatchery and nursery from which they originated. PMID- 7482472 TI - [What are the rules?]. PMID- 7482473 TI - [Revision evaluation of feed for horses]. PMID- 7482474 TI - [Rupture of the follicle]. PMID- 7482475 TI - [Veterinary dentistry (11). Feline gingivitis-stomatitis-pharyngitis complex. Chronic/recurrent stomatitis in cats]. AB - This is the fourth article in a series on veterinary dentistry in cats. This article describes the clinical signs, possible investigations, and differential diagnosis of the gingivitis-stomatitis pharyngitis complex (GSP complex), a complex and frequently occurring disease. Strategies for the treatment of feline chronic stomatitis complex, which is frequently idiopathic, are presented. PMID- 7482476 TI - [Veterinary medicine after 2002: primary and continuing education]. PMID- 7482477 TI - [The veterinarian after the year 2002]. PMID- 7482478 TI - [Control of Salmonella on dairy farms]. PMID- 7482479 TI - [Half-yearly Mastitis Panel Meeting. Management protocol appears to be a useful weapon against mastitis]. PMID- 7482480 TI - [Purchase of sheep and the BVD-risk connected with it]. PMID- 7482481 TI - ['Oat bumps' in horses. Differential diagnosis and nutritional aspects]. AB - Small bumps or swellings of about 0.5 mm in diameter in the skin of horses are called 'haverbultjes' in the Netherlands. Comparable skin diseases described in the literature are 'sweet feed-, protein- and heat bumps'. The differential diagnosis and the possible nutritional causes of 'haverbultjes' are presented. PMID- 7482482 TI - [Tendovaginitis of the musculi biceps brachii in dogs]. AB - Successful surgical correction of bicipital tenovaginitis and extensive new bone formation in the shoulder joint of an 8-year-old dog is described. Radiography and computed tomography revealed severe arthrosis of the shoulder joint with osteophyte formation encircling the bicipital tendon in the intertubercular groove. The diagnostic approach and therapy are described and discussed with reference to recent literature. PMID- 7482483 TI - [Annual report Veterinary Disciplinary Tribunal and Professional Board 1994. Attention to communication and reception of complaints is necessary]. PMID- 7482484 TI - [Drs. A.H.H.M. Mathijsen is leaving the Library. Already user-friendly before the term was coined]. PMID- 7482485 TI - [What is your diagnosis? Paratuberculosis]. PMID- 7482486 TI - [Testing area Wehl: study of risks of contacts in behalf of foot-and-mouth disease]. AB - A study on contacts between livestock holdings was conducted in a geographically defined area of 4 by 6 km in the Netherlands. The farmers were asked to record all contact on and off farm during a period of 2 weeks. The number of contacts in the 2 week period was high, on average 91.8 contacts per farm. The risk of spreading foot-and-mouth disease on or off the farm was greater for cattle farms than for swine farms. Relatively more people had contact with the animals while visiting cattle farms, thereby causing the higher risk. Most contacts occurred over short distances. Almost 50% of the contacts were within the research area. This finding confirms the importance of restricted areas around farms with foot and-mouth disease infections. PMID- 7482487 TI - [Sterin accreditation for the Inspection and Control activities of the Health Service. GD-IC: a new branch on the GD-tree! (Health Service)]. PMID- 7482488 TI - [Pioneers: veterinarians from earlier times (12). Pieter van Naaldwijck and Peter Janz Almanus van Coer]. PMID- 7482489 TI - [Certification]. PMID- 7482490 TI - [Complications following perineal urethrostomy in the male cat]. PMID- 7482491 TI - Nomenclature for factors of the HLA system, 1995. PMID- 7482492 TI - Anchored PCR cloning of the novel HLA-Cw*0704 allele detected by PCR-SSP. AB - The novel HLA-Cw*0704 allele, previously detected as the PCR-SSP variant Cw7/8v, has been cloned and sequenced from the homozygous typing cell KRO3/4 after amplification by anchored PCR. The nucleotide sequence of Cw*0704 is closely related to those of other Cw*07 alleles, but carries specific changes in exon 3 consistent with its serological behavior-a short Cw7 cross-reactive with antibodies directed against HLA-Cw8. Some of the substitutions of Cw*0704 have not been previously described for HLA-C but are found in HLA-B alleles and in published C sequences of non-human primates. The new allele carries a novel polymorphism in its 5' untranslated region (5' ut) that could be shared by all Cw*07 alleles. By PCR-SSP, Cw*0704 is a relatively common allele in English Caucasoids at a frequency of 4.6%. It is most often observed on HLA-B44 haplotypes previously described as HLA-C "blank", although linkage disequilibria with other HLA-B specificities have been found. PMID- 7482493 TI - The relative distribution of B35 alleles and their IEF isotypes in a HLA-B35 positive population. AB - The HLA-B35 serotype represents a group of antigens detectable by IEF, cytotoxic T cells, and by sequencing analysis. Four isotypes and eight alleles have been thus far reported. We have determined the relative frequencies of these B35 subtypes in a group of 203 unrelated people. Dot blot hybridization of PCR amplified products was performed using 23 sequence-specific oligo probes designed based on the EMBL HLA class I sequence database. The amplification was achieved by a pair of group-specific primers, producing approximately 600 bp fragments. By hybridization pattern analysis, we found that four alleles represent over 95% of the B35+ population, with relative frequency of 48.2% for B*3501, 23.7% for B*3502, 15.2% for B*3503, and 8.0% for B*3508. We also identified 3 individuals with B*3504 and one with B*3505, and seven samples with new patterns. B*3501 and B*3503 exactly correlated with the most common isotype B35.3, B*3502 and B*3504 with B35.2, B*3508 may be the B35.1 IEF isotype. The B*3505 was identified from an individual with B35 IEF variant form. Our study shows that the B35 antigen has a wide distribution of alleles, and that many more B35-related alleles may yet to be uncovered. PMID- 7482494 TI - CD50 (intercellular adhesion molecule-3) is expressed at higher levels on memory than on naive human T cells but induces a similar calcium mobilization on both subsets. AB - CD50, the intercellular adhesion molecule-3 (ICAM-3), is expressed almost exclusively on hematopoietic cells. T lymphocytes display a bimodal distribution on CD50 expression levels. It was observed that CD45RO+ cells expressed higher levels of CD50 than CD45RA+ T lymphocytes. A similar situation was observed when CD4 and CD8 subpopulations were analyzed, with CD8+ cells expressing higher levels of CD50 than CD4+ cells. When adult T lymphocytes were analyzed by three color flow cytometry in CD8+CD45RA+ cells both CD50low and CD50high expressing cells were detected, in accordance with several memory markers on T lymphocytes, whereas only cells with a low level of CD50 were observed in CD4+CD45RA+. The different level of CD50 expression was confirmed by analyzing purified CD45RA+ and CD45RO+ T cells. Moreover, after the comparison of CD50 expression level in thymocytes, cord blood and adult T lymphocytes, a progressive increase was observed. When T cells were sorted by their intensity of CD50 expression, only CD50high cells proliferated in response to tetanus toxoid. Therefore, the phenotypic and functional analysis of adult and cord blood T lymphocytes as well as thymocytes indicates that CD50 expression increases during the maturation process of T lymphocytes: from the lowest CD50 levels present on CD1+ thymocytes, to the highest levels of CD50 on human memory T cells. In addition, we have observed that after CD50 cross-linking on human T lymphocytes, a transient increase in intracellular calcium concentration ([Ca2+]i) is produced. When CD45RA+ and CD45RO+ T cells were analyzed, in spite of the level of CD50 expression, the stimulation through CD50 induced a similar level of Ca2+ mobilization in both subpopulations, contrasting with the higher rise in [Ca2+]i induced by CD3 stimulation on CD45RA+ versus CD45RO+. These data suggest that the signal transduction pathways activated after CD50 cross-linking are, at least partially, independent of those involved after CD3 stimulation. PMID- 7482495 TI - Increased surface expression of class I MHC molecules on immunogenic cells derived from the xenogenization of P815 mastocytoma cells with 8-methoxypsoralen and long-wavelength ultraviolet radiation. AB - In a previous study we demonstrated that the treatment of the highly tumorigenic cell line, P815, with 8-methoxypsoralen and long-wavelength ultraviolet radiation resulted in the production of several immunogenic clones (tum-). Mice inoculated with the tum- cells survived much longer than mice inoculated with the original tumorigenic cells (tum+). It was suggested that the increased survival of mice treated with the tum- clones arose as a result of an increased antigenicity derived from the phototreatment. In this report we show that the tum- cells have a greater density of class I MHC molecules on their surface (50-157% compared to P815). Class I MHC density on the cell surface is required to elicit targeted cytotoxic responses. These results can be considered in terms of human class I MHC assays which show that many human tumor cells have a reduced expression of class I MHC. Because other DNA damaging agents have also been shown to enhance class I expression, it is suggested that in addition to the cytotoxic effects of these agents, other pleiotropic effects must be considered. Photochemotherapy may phenotypically alter cells so that the enhanced expression of class I MHC molecules on the surface of phototreated cells may be associated with the clinical responses observed in cutaneous T cell lymphoma patients. PMID- 7482496 TI - On the nucleotide sequences of B*2702 and B*2705. PMID- 7482498 TI - Genetic polymorphisms of the TNFB and HSP70 genes located in the human major histocompatibility complex in sarcoidosis. PMID- 7482500 TI - Two novel DRB1 alleles, DRB1*1118 and DRB1*1119, detected by PCR-SSOP and confirmed by DNA sequencing. PMID- 7482497 TI - A novel HLA-A24 allele (A*2405) identified by single-strand conformation polymorphism analysis and confirmed by solid-phase sequencing and isoelectric focusing. PMID- 7482499 TI - Steroid 21-hydroxylase gene polymorphism in Addison's disease patients. PMID- 7482501 TI - Discrepant sequence at codon 57 of DQB1: implication on HLA typing of "Asp 57" in I.D.D.M. PMID- 7482503 TI - Determination of HLA class II alleles by genotyping in a Manchu population in the northern part of China and its relationship with Han and Japanese populations. AB - The genetic polymorphism of the HLA class II loci was investigated in a Manchu population resident in the northern part of China and compared with those of other Asian populations including Japanese and Han. In 8 DQA1 alleles, the most frequent allele was DQA1*03 with the gene frequency of 25.5%. Of 15 DQB1 alleles tested, 11 were observed and the most common allele was DQB1*0301 with the gene frequency of 24.5%. Among 19 DPB1 alleles, 11 were detected and DPB1*0501 (43.8%) was the most frequent allele as observed in other Asian populations such as Japanese, Chinese and Korean. Of 43 DRB1 alleles tested, 21 were detected and DRB1*0901 (14.0%), *1501 (11.0%), *1201 (11.0%), *07 (9.0%) and *1401 (9.0%) were highly predominant and account for the high frequencies of DR9, DR2, DR5, DR7 and DR6. In the DRB3 gene (DR52), DRB3*0202 (18.0%) was the most frequent. With respect to the DRB4 gene (DR53), the gene frequency of DRB4*0101 was 35.0%. Of 3 DRB5 alleles detected, DRB5*0101 (11.0%) was highly predominant. Comparison of HLA class II allele frequencies in Manchu with those in Japanese and Han Chinese populations (South & North) detected some significant differences and genetic divergence between these Oriental populations. The dendrogram constructed by the neighbor-joining (NJ) method based on the allele frequencies of DQA1, DQB1, DPB1 and DRB1 of 10 representative populations over the world suggested that Manchu is the closest, but at the same genetic distance to both Northern and Southern Han Chinese. PMID- 7482502 TI - Comparative analysis of HLA polymorphism at the serologic and molecular level in Moroccan and Ashkenazi Jews. AB - The Jewish people comprise two major groups, one encompassing the Jews of Ashkenazi (Central and Eastern European) origin and the other including those of Sephardic (Middle Eastern and North African) descent. To the latter belong the Jews of Moroccan stock, who form the largest Jewish subgroup among the non Ashkenazi population living in Israel. As the members of each of these groups differ in physiognomy and life style, it was of interest to investigate whether these differences are also reflected in their respective HLA compositions. To this end, 132 subjects of Ashkenazi and 113 individuals of Moroccan origin residing in Israel were tested and the results compared with data for other populations made available by the 11th International Histocompatibility Workshop. Comparison between their HLA profiles and those of non-Jews revealed that the Jewish groups in some aspects resembled one another but in others showed disparities. The dissimilarities between the various groups are expressed in terms of gene and haplotype frequencies, as well as in HLA-disease associations (as for example rheumatoid arthritis, erosive lichen planus, primary Sjogren's syndrome, pemphigus vulgaris). However, both Jewish groups shared some unique features with respect to HLA class II allelic frequencies, pointing to a common ancestry. PMID- 7482504 TI - Increased serum concentration of soluble HLA-DR antigens in HIV infection and following transplantation. AB - HLA class I and class II antigens circulate in serum as soluble molecules. Increased concentrations of soluble HLA class I molecules have been demonstrated in viral diseases, in rejection episodes following organ transplantation and in graft versus host disease. To explore the possibility of a variation of the serum concentrations of soluble HLA class II molecules in the same pathologic conditions we developed a double determinant immune assay that detects whole soluble HLA-DR molecules (sHLA-DR). The mean level of sHLA-DR antigens in sera from 23 healthy individuals was 0.64 +/- 0.72 microgram/ml. Elevated serum concentrations of sHLA-DR molecules were detected in sera from HIV infected patients in CDC2/3 and in CDC4 C1 stages (2.0 +/- 1.7 micrograms/ml and 4.6 +/- 1.7 micrograms/ml, respectively), in sera from patients affected by acute rejection after liver transplantation (5.3 +/- 3.7 micrograms/ml) and in sera from patients affected by severe acute graft versus host disease following bone marrow transplantation (8.8 +/- 3.1 micrograms/ml). The increase of sHLA-DR molecules in these sera significantly correlated with the elevation of soluble HLA class I antigens (P = 0.0004). The reported data suggest that both soluble HLA class I and class II molecules serum levels increase during viral infections and strong immune reactions and could suggest the involvement of these molecules in immunoregulation. PMID- 7482507 TI - HLA class II haplotype studies bring molecular evidence for population affinity between Madagascans and Javanese. PMID- 7482506 TI - Yet another novel HLA DRB1 allele (DRB1*1317) and its misidentification by PCR SSP. PMID- 7482505 TI - A new HLA-DRB1*1116 allele sharing DR13 and DR11 sequence motifs. PMID- 7482508 TI - Polymorphism in the LMP2 and LMP7 genes and adult rheumatoid arthritis: no relationship with disease susceptibility or outcome. PMID- 7482509 TI - Heat shock protein 70 gene polymorphisms and multiple sclerosis. PMID- 7482510 TI - Nomenclature for factors of the HLA system, update June 1995. PMID- 7482511 TI - A comprehensive approach for typing the alleles of the HLA-B locus by automated sequencing. AB - We describe an approach for typing alleles of the HLA-B locus by using automated sequencing technology. The exon 2 and exon 3 nucleotide sequence of each allele is determined directly from genomic DNA in two steps. In the first step, HLA-B exon 2, intron 2 and exon 3 sequences are amplified with one or two primer pairs out of a panel of 5 primer pairs that describe all known HLA-B alleles. In the second step, templates are sequenced in 5' and 3' orientations in a PCR assay that utilizes Taq polymerase to incorporate fluorescent dye-labeled nucleotides into each new strand synthesized. Gel electrophoresis of the labeled products is performed in an automated DNA sequencer. The derived sequences are aligned against reference sequences and each nucleotide position is evaluated for homology to consensus sequence. Using this strategy, the HLA-B allele sequence is directly ascertained with precision and efficiency. The automated sequencing strategy can be readily applied in the clinical laboratory as a practical tool for high resolution typing of HLA-B alleles. PMID- 7482513 TI - An evaluation of a multicenter study on HLA-DPB1 typing using solid-phase Taq cycle sequencing chemistry. AB - In HLA Class II genes, polymorphism is mainly located in the second exon. Most DNA based typing methods are confined to the identification of specific sequence motifs in the second exon. In contrast, Sequencing Based Typing (SBT) elucidates the entire exon 2 sequence for typing. Comparison of the obtained exon 2 sequence with an allele sequence library results in allele assignment. We tested the applicability of SBT using a protocol for amplification followed by solid phase Taq-cycle sequencing for HLA-DPB1 typing. A panel of 32 samples were typed by SBT at five test sites which are participating in the Sequencing Based Typing component of the 12th International Histocompatibility Workshop. The panel represents the existing polymorphism at all known polymorphic positions of exon 2, both in homozygous and heterozygous combinations. In this multicenter study we focused on the reliability of analyzing heterozygous sequences for HLA typing. A multi-sequence analysis approach, Polall, was developed to evaluate sequences obtained. The assignment of homozygosity and heterozygosity was validated by cluster analysis of chromatographic data of all sequences. Sequence characteristics were examined and considered for appropriate assignment. Differences in sequence characteristics that occurred between the test sites are considered in detail. The evaluation of data of 5 test sites reveals that Taq cycle sequencing can reliably be performed for HLA-DPB1 SBT. PMID- 7482514 TI - Clinical significance and cost-effectiveness of 24-hour ambulatory blood pressure monitoring. AB - Ambulatory blood pressure (BP) monitoring as an adjunct to casual/clinic BP measurements is currently used widely for the diagnosis and treatment of hypertension. It has been established that ambulatory BP monitoring is essential to confirm "white coat" hypertension, drug-resistant hypertension, duration of drug action, short-term BP variation, and nocturnal and on-the-job BP levels. It is estimated that approximately 10,000 ambulatory BP monitoring devices are currently used in Japan. That number would increase if 1) a standard algorithm with a theoretical basis to determine BP levels is introduced for ambulatory BP monitoring devices based on cuff-oscillometric method, 2) the reproducibility of ambulatory BP levels is confirmed, 3) reference values for evaluating ambulatory BP monitoring levels are established, and 4) the clinical significance and prognostic value of ambulatory BP monitoring is established. If such problems is settled, the use of ambulatory BP monitoring in the diagnosis and treatment of hypertension would be national health insurance and would improve the prognostic accuracy of evaluating hypertension as well as the cost-effectiveness of screening, diagnosis and treatment of hypertension. PMID- 7482512 TI - Complete subtyping of the HLA-A locus by sequence-specific amplification followed by direct sequencing or single-strand conformation polymorphism analysis. AB - A variety of reasons related to the HLA class I system has complicated the application of molecular approaches to HLA class I typing. Here we present a PCR based HLA-A typing strategy considering the sequence variations of the two most polymorphic exons which allows complete subtyping of the HLA-A locus. The method is based on a sequence-specific amplification identifying the serologically defined HLA-A specificities. The PCR products generated by these group-specific primers bear the sequence information necessary for a postamplification specificity step. The primer pairs are located within one exon, either exon 2 or exon 3, which avoids amplification of polymorphic intron sequences allowing subsequent single-strand conformation polymorphism analysis and facilitating direct sequencing. Using this method we investigated 48 cell lines and 153 clinical samples. 23 PCR reactions are performed per individual for the assignment of the serological specificities A1-A80. The reproducibility was 100% in all cell lines and 85 clinical samples typed on two separate occasions. With the exception of 13 out of 231 possible serological combinations all homozygous and heterozygous combinations of A1-A80 can be distinguished by specific amplification patterns. Comparing the PCR based typing results with those of serology in 12% a discrepancy was found. Solid-phase sequencing or SSCP analysis of the group-specific PCR fragments allowed complete subtyping of the HLA-A locus. This strategy can identify all 48 HLA-A alleles based on the sequence variations of the 2nd and 3rd exon. 1128 homozygous and heterozygous allele combinations are possible for the HLA-A locus. Only 4 out of these 1128 allele combinations remained unresolved. PMID- 7482515 TI - Effects of human placental extract on brain monoamines and monoamine oxidase activity in rats. AB - Human placental extract, an agent clinically used world-wide in a number of physiological anomalies, has been claimed to be effective in children of slow learners. Since the monoaminergic neurotransmitter systems in the brain play an important role in the processes of learning and memory, we examined the effects of human placental extract on the levels of norepinephrine, dopamine and serotonine in rat brain as an attempt to evaluate the possible underlying biochemical mechanism of action of the extract. We also determined the changes of brain monoamine oxidase (MAO) activity following placental extract treatment. The results showed that subchronic (5, 10, 15 or 20) administration of placental extract (2-4 ml/kg/day) had the effect of increasing all the monoamines and decreasing the MAO activity which could be the possible mode of action of the extract in slow learners. PMID- 7482516 TI - Daily single-dose regimen and alternate-two-week triple-dose/day regimen of oral ofloxacin for the prophylaxis and control of exacerbations of chronic respiratory tract infections. AB - Two different oral ofloxacin regimens, a daily single-dose regimen with 200 mg (Regimen I) and an every-two-week multidose regimen with 3 x 200 mg/day (Regimen II) was compared as to the efficacy in controlling repeated acute exacerbations of chronic respiratory tract infections. Fifty-eight patients consisting of 19 patients each of bronchiectasis and pulmonary emphysema, 10 patients of chronic bronchitis, 5 patients of old pulmonary tuberculosis, 4 patients of diffuse panbronchiolitis and 1 patient of multiple pulmonary bullae were evaluated: 32 patients in Regimen I and 26 patients in Regimen II. The corrected mean incidence of exacerbations per case decreased from pre-study 2.47 to intra-study 0.59 in Regimen I, and from pre-study 2.66 to intra-study 0.95 in Regimen II, respectively, with a statistically significant difference (p < 0.05, respectively). Only one of 12 persistent isolates of Pseudomonas aeruginosa acquired a certain degree of resistance to ofloxacin. Adverse reactions were found in six of 66 patients. We conclude that long-term administration of an new quinolone, especially a daily single-dose regimen with ofloxacin, is useful to control acute exacerbations of chronic respiratory tract infections. PMID- 7482517 TI - Proximal tibiofibular joint: a radiographic and computed tomographic study. AB - In 671 knees of 351 patients, the proximal tibiofibular joint was radiologically investigated. To clearly detect the contour of the joint, radiographs were obtained under fluoroscopic control with the lower extremity internally rotated. The contour was classified into three types: the joint lines of both tibia and fibula are straight (A); the joint line of the fibula is convex (B); the joint line of the tibia is convex (C). The rotation and the inclination of the joint were measured. Of the 671 knees, CT examination was also carried out in 51 knees. The obliquity of the joint was measured and the location of the fibula as related to the tibia was evaluated by CT. We also examined the relationships between the contour and the rotation, inclination, obliquity of the joint and the location of the fibula, and determined the characteristics of type B and C as compared with type A. The characteristics of type B were as follows: (1) this type was mainly seen in younger patients, (2) the rotation was the lowest and the inclination was the highest among the three types, (3) in CT the joint line was more saggital and the fibula was located more anteriorly. The characteristics of type C were as follows: (1) this type was seen in older patients, (2) the rotation was the highest and the inclination was the lowest among the three types, (3) in CT the joint line was more frontal and the fibula was located more posteriorly. PMID- 7482518 TI - Regional lung function following upper sleeve lobectomy for bronchogenic carcinoma. AB - Regional lung function of the operated side following upper sleeve lobectomy (n = 8) and simple upper lobectomy (n = 7) for lung cancer was evaluated. Regional ventilation was studied with Krypton 81m and regional pulmonary blood flow was studied with Technetium 99m. Measurements were taken from 12 to 24 months after operation. The ventilation rate of the operated side following right upper sleeve lobectomy (n = 5) was 42.9 +/- 6.7% and the perfusion rate was 37.4 +/- 4.6%. The regional ventilation rate of the operated side after simple right upper lobectomy (n = 3) was 45.9 +/- 10.5% and the perfusion rate was 46.2 +/- 5.2%. For the patients with left upper sleeve lobectomy (n = 3), these ratios were 29.9 +/- 11.1% and 19.2 +/- 3.6%, respectively. For the patients with simple left upper lobectomy (n = 4), these ratios were 27.3 +/- 4.5% and 22.3 +/- 3.3%. There were no statistical differences between the group with upper sleeve lobectomy and that with simple upper lobectomy. Regional function improved gradually with time. In conclusion, the regional lung function of the operated side following sleeve lobectomy was well preserved and showed no difference when compared with the regional function after simple lobectomy for lung cancer. PMID- 7482519 TI - Effect of motilin on colonic motor activity in the interdigestive state in conscious dogs. AB - The aim of the present study was to investigate the effect of motilin at various doses on colonic motility in the interdigestive state. Colonic motility was investigated in five dogs equipped with strain gauge force transducers on the gastric antrum, and on the proximal, middle, and distal colon. Exogenously infused motilin (0.2, 0.5, 1.0 and 2.0 micrograms/kg-hr) dose-dependently increased colonic motility, but the doses of motilin that significantly enhanced colonic motility were 1.0 and 2.0 micrograms/kg-hr in all areas of the colon. Motilin at 0.2 microgram/kg-hr increased the plasma motilin concentration to almost equal to the physiological peak values. The excitatory effect of motilin (2.0 microgram/kg-hr) was abolished by atropine but was not affected by hexamethonium. These results indicate that exogenously infused motilin, which increased plasma motilin concentration to above the physiological peak level, stimulated colonic motility by affecting postsynaptic cholinergic neurons. PMID- 7482522 TI - Gadolinium-enhanced magnetic resonance imaging of meningeal carcinomatosis in colon cancer. AB - Meningeal carcinomatosis has been infrequently reported in colorectal carcinoma. Recently the value of gadolinium-enhanced magnetic resonance imaging (MRI) in the examination of this disease has been described. We report a 55-year-old man with meningeal carcinomatosis from a known colon cancer. In this case, a gadolinium enhanced MRI of the brain showed enhancing dura and nodular lesions which were not detected by enhanced computed tomography. The gadolinium-enhanced MRI may serve as an indicator of leptomeningeal involvement and may be a useful test in addition to the evaluation of cerebrospinal fluid cytology and chemistry. PMID- 7482521 TI - Multivariate analysis of limb long bone growth during the human prenatal period. AB - Prenatal development of the human limb bones (humerus, ulna, radius, femur, tibia, fibula) was studied. Cross-sectional data, consisting of 21 anthropometric measures of these bones were collected from 122 Japanese fetuses between the gestational ages of 18 and 40 weeks. Principal component analysis was applied to the data to provide multivariate assessments of morphological patterning among the variables. Three orthogonal components that accounted for 94.2% of the overall sample variation were extracted. The first component accounted for 88.2% of the variation and represented an axis of overall body size that was dependent on gestational age. The second and third components both reflected a trend in shape involving the cartilaginous parts of the humerus and femur. The findings indicated that different growth factors existed between the ossified and cartilaginous parts of fetal long bones. Multivariate allometric coefficients were extracted from the first principal component. The variables that were related to the construction of the articulate showed positive allometry, and the central widths of the diaphysis showed negative allometry compared with total size. In prenatal skeleton, certain shape changes are functionally linked to and required by changes in body size. A comparison among the coefficients of long bone length revealed that lower limb bones grew faster than upper limb bones during the period under study here. Allometric coefficients were equivalent among bones within a limb, whereas homologous bones in the upper and lower limb grew at different rates. PMID- 7482520 TI - Comparison of serum and urinary levels of modified nucleoside, 1-methyladenosine, in cancer patients using a monoclonal antibody-based inhibition ELISA. AB - To evaluate the clinical usefulness of serum 1-methyladenosine, several modifications have been made in our previously established inhibition ELISA system. Horseradish peroxidase (HRP) labeled anti-mouse IgG and 3,3',5,5' tetramethylbenzidine (TMBZ) were used as a secondary antibody and a substrate, respectively. The second blocking was done just before the addition of the secondary antibody. The standard curve of the modified ELISA system showed good linearity between 1 and 1,000 ng/ml, and the detection limit was 50 pg/well. Using the ultrafiltrated-serum samples, serum 1-methyladenosine levels in healthy individuals and cancer patients were determined. The mean level of 1 methyladenosine in 31 healthy individuals was 28.3 +/- 7.9 ng/ml, and cut off value was set at 44.1 ng/ml (Mean+ 2SD). In cancer patients, elevated levels of serum 1-methyladenosine above the cut off value were detected in 4 out of 25 cases tested, though 11 cases had elevated urinary 1-methyladenosine levels above the cut off value (3.23 nmol/mumol creatinine). Since 1-methyladenosine has no interaction with serum proteins and its molecular weight is quite low, it might be rapidly excreted into the urine. PMID- 7482523 TI - Gastric mucosal phosphatidylcholine hydroperoxide increases during cold water immersion restraint stress in rats. AB - To know the role of lipid peroxidation in the pathogenesis of stress-induced gastric mucosal lesions, we performed the present study. Both gastric mucosal lesions and gastric mucosal phosphatidylcholine hydroperoxides (PCOOH) levels during cold water-immersion restraint (CWIR) stress were measured by the chemiluminecence-HPLC method in Sprague-Dawley rats. Ulcer indices increased time dependently during the stress. Gastric mucosal PCOOH levels also significantly increased and reached a peak level (250% of untreated level) after 2 hr of stress. These results suggest that lipid peroxidation may play an important role in the pathogenesis of gastric mucosal lesions induced CWIR stress. PMID- 7482524 TI - Changes in nitric oxide synthase activities in the cerebellum during development and aging of C57BL/6 mice. AB - Nitric oxide (NO) is one of the neurotransmitters in the cerebellum. Activities of NO synthase and densities of granule cells were determined in the cerebellum in an attempt to elucidate a mechanism of biochemical changes on the anatomical basis during aging. Activities of Ca2+/calmoduline dependent NO synthase were measured in the cerebellum in male and female mice, from 18 gestation days to 28 months of age. There were significant differences in NO synthase activities among the groups of prenatal (18 gestation days), middle aged (5-18 months) and old aged (21-28 months) mice. Between groups of old and middle ages, a significant decrease with aging was found in the mean density of granular cells, which express NO synthase. Changes in NO synthase activities and granule cell densities occurred to a similar degree between old and middle ages. The present data suggest that an age-associated rise and fall of its activities likely occur in parallel with those of granule cell densities, and thus may represent impairment of cerebellar function during aging. PMID- 7482525 TI - A method of short-term cryostorage and selection of viable sperm for use in the various assisted reproductive techniques. AB - The objective of this study was to determine if spermatozoa, following short-term cryostorage at 5 degrees C in Test-Yolk buffer (TYB; ZBL, Inc., Lexington, KY, USA), could be recovered and improved via the SpermPrep filtration method and to assess the possibly enhanced fertilizing capacity of the selected spermatozoa. Semen specimens from 20 men were collected, evaluated, diluted 1:1 (v/v) with TYB, divided into aliquots and cooled to 5 degrees C for 24 and 48 hr. Semen samples were assessed for volume, sperm count, percentage and grade of motility, percentage of morphologically normal spermatozoa and outcome of the sperm penetration assay (SPA). After storage, aliquots were rewarmed at 37 degrees C, centrifuged, and the pellet was resuspended in 1.0 ml of SpermPrep media (ZBL, Inc.). Following 15 min of incubation, the rewarmed spermatozoa were filtered via the SpermPrep I filtration column (ZBL, Inc.) and assessed accordingly. The results obtained in this study indicate that the short-term cryostorage procedure yielded spermatozoa of adequate qualitative characteristics when compared to the fresh spermatozoa. Furthermore, filtration of rewarmed specimens yielded spermatozoa of significantly higher qualitative characteristics and superior fertilizing capacity following a short-term cryostorage period in TYB when compared to fresh and rewarmed spermatozoa (p < 0.05). This method of short-term cryostorage in TYB and selection of superior spermatozoa via the SpermPrep filtration method could further enhance the fertilizing ability of patients who produce spermatozoa characterized by deficient capacitation, acrosome reaction and subsequent fertilization. PMID- 7482526 TI - Sr2+ can pass through Ca2+ entry pathway activated by Ca2+ depletion, but can be hardly taken up by the Ca2+ stores in the rat salivary acinar cells. AB - When Sr2+ was introduced to the external solution after the depletion of Ca2+ from stores of submandibular acinar cells by ACh stimulation, Sr2+ entered cytoplasm of the cell, like the case of Ca2+. SK&F 96365, a Ca2+ channel blocker, or Ni2+ blocked this divalent cation entry. Sr2+ entering the cell continued to increase to a steady level, after the cessation of stimulation, when Sr2+ was present in the external solution, unlike the case of Ca2+. Ca2+ entered cells which had been stimulated with ACh in Sr(2+)-containing external solution. In the cells to which Sr2+ has been applied after the store depletion with ACh, Sr2+ cannot be released by the renewed ACh stimulation, unlike the case of Ca2+. 89Sr2+ uptake by the parotidic microsomal fraction 100 min after addition of ATP was 15.08 +/- 0.70 nmol/mg protein, whereas 45Ca2+ uptake was 144.19 +/- 16.93 nmol/mg protein. It was concluded that in the salivary acinar cells Sr2+ can be a substituent for Ca2+ in the mechanism of entry from the extracellular fluid but cannot be in the mechanism of uptake into the stores. PMID- 7482527 TI - Dementia and physical disability as competing risks for mortality in a community based sample of the elderly Japanese. AB - To examine whether an excess mortality due to dementia is independent of coexisting physical disability, a probability-sample of the non-institutionalized elderly (n = 3,308) living in Sendai City, Japan was followed between 1988 and 1991. Of those, 128 were diagnosed as dementia in 1988 by psychiatrists, using Diagnostic and Statistical Manual of Mental Disorders, Third Edition-Revised as a diagnostic standard. Information on the ability to perform activities of daily living (ADL) was collected by self-report of the study subjects in 1988 baseline survey. The survival status was investigated three years later. The risks of dementia and co-existing ADL disability for mortality was examined by Cox proportional hazard models. The results indicated that the relation between dementia and mortality was two-fold, depending upon the physical functions. Dementia increased the risk for mortality among those without ADL disability, but it did not so among those with ADL disability, rather ADL function was a stronger predictor for mortality among the latter individuals. Prevention and treatment of physical disability would be important for improving the survival of the demented people. PMID- 7482528 TI - Intestinal toxicity of acrylonitrile: in vitro metabolism by intestinal cytochrome P450 2E1. AB - Acrylonitrile (VCN) is known to cause extensive gastrointestinal damage and tumors in rats. In this study the metabolism of VCN to cyanide (CN-) was characterized in the small intestinal mucosa. The majority of the metabolic reactivity was localized in the microsomal fraction and required reduced nicotinamide adenine dinucleotide phosphate for maximal activity. The intestinal metabolism of VCN to CN- was characterized with respect to VCN concentration, time, pH, and microsomal protein concentration. VCN metabolism to CN- was enhanced significantly by the addition of sulfhydryl compounds such as glutathione, cysteine, and D-penicillamine (10 mM) to 142, 161, and 189% of control, respectively. The intestinal bioactivation of VCN to CN- was enhanced by microsomes obtained from intestinal mucosa of phenobarbital (455% of control), beta-naphthoflavone (375% of control), 4-methylpyrazole (305% of control), or ethanol (165% of control)-treated rats. Addition of ethanol (80 mM) to incubation mixtures containing control or ethanol-induced microsomes resulted in significant inhibition of microsomal metabolism of VCN to CN- to 20 and 34% of control, respectively. Addition of dimethyl sulfoxide induced a similar inhibitory effect on VCN metabolism by control or ethanol-induced microsomes (8 and 26% of control, respectively). Furthermore, antibody to cytochrome P450 2E1, but not antibody to cyt P450 2B1, significantly inhibited VCN metabolism by ethanol-induced intestinal microsomes to about 25% of control. Mild inhibition (80-85% of control) of VCN metabolism was detected when antibody to cyt P450 2B1 or 2E1 was added to incubation mixtures containing Pb-induced intestinal microsomes. These findings indicate that extrahepatic tissues such as the intestinal mucosa are capable of metabolizing VCN to CN- and establish a major role of intestinal cyt P450, particularly cyt P450 2E1, in the intestinal metabolism of VCN to CN-. PMID- 7482529 TI - Stressor-specific activation of heat shock genes in H35 rat hepatoma cells. AB - Responses of Reuber H35 hepatoma cells exposed to increasing doses of heat, arsenite ions, or cadmium ions were investigated. Doses which are capable of activating the heat shock transcription factor (HSF) were determined. The activity of this factor in the poststress period was correlated to the recovery of total protein synthesis in this same period. Subsequently, increases of mRNA levels and rates of synthesis of heat shock protein (HSP)60, HSP68, and HSP84 after exposure to the stressors were determined. We generally found that the rate of HSP synthesis correlated well with HSP mRNA levels, supporting the idea that the stress response is regulated mainly at the transcriptional level. A stressor specific pattern of HSP mRNA induction can be observed. The most striking example is cadmium chloride which does not induce HSP60 nor its mRNA. Interestingly, under these conditions maximal activation of HSF is observed. Therefore, we conclude that more processes than just HSF activations are involved in the induction of heat shock genes. PMID- 7482530 TI - Detoxification of vinyl carbamate epoxide by glutathione: evidence for participation of glutathione S-transferases in metabolism of ethyl carbamate. AB - Vinyl carbamate epoxide (VCO) is believed to be the metabolite of ethyl carbamate (EC) ultimately responsible for its carcinogenic effects. This study investigates the role of glutathione (GSH) in protection against VCO-mediated adduct formation, and the involvement of glutathione S-transferases (GSTs) in detoxification of VCO. Formation of 1,N6-ethenoadenosine from VCO and adenosine in vitro was employed as a measure of VCO toxicity. GSH inhibited formation of ethenoadenosine in a concentration-dependent manner at concentrations ranging from 1 to 8 mM. This effect was significantly enhanced by addition of rat liver GST. Mouse liver cytosol was also found to inhibit formation of ethenoadenosine in a concentration-dependent manner, and the inhibition was relieved by addition of S-octylglutathione, a competitive inhibitor of GST. Pretreatment of mice with 1% dietary (2(3)-tert-butyl-4-hydroxyanisole (BHA) caused parallel increases in cytosolic GST activity and cytosolic enhancement of detoxification of VCO by GSH. Furthermore, BHA increased hepatic steady-state concentrations of GSH greater than twofold. The effect of BHA on detoxification of EC in vivo was examined using formation of 2-oxoethylvaline (OEV) adducts of hemoglobin as a biomarker. Pretreatment with BHA decreased overall formation of OEV adducts 23%. The major conclusions of this study are (1) VCO can be detoxified by spontaneous conjugation with GSH, (2) conjugation of VCO with GST can be catalyzed by GST(s), (3) pretreatment with BHA protects against binding of active EC metabolites in vitro and in vivo, and (4) the protective effect of BHA against EC is mediated by increases in GST activity and GSH concentration. PMID- 7482531 TI - Dose- and route-dependent alteration of metabolism and toxicity of chloroform in fed and fasting rats. AB - Effects of overnight food deprivation on the metabolism and toxicity of chloroform (CHCl3) administered to rats per os (po), intraperitoneally (ip), or by inhalation (inh) at different doses were investigated. Rats that had been either deprived of food overnight or normally fed were challenged with CHCl3 either po (0, 100, 200, or 400 mg/kg), ip (0, 100, 200, or 400 mg/kg), or inh (0, 50, 100, or 500 ppm for 6 hr). Overnight fasting increased CHCl3 metabolism in vitro about threefold with a decrease of liver glutathione content to 67%. The fasting caused route- and dose-dependent alteration in the metabolism and toxicity of CHCl3. The area-under-the-curve (AUC) of blood CHCl3 concentration was invariably smaller following po than ip administration, and CHCl3 administered po caused more severe hepatic damage than did the same amount of CHCl3 administered ip. With po administration, the AUC (toxicity) of CHCl3 in fasting rats was significantly smaller (higher) than that of fed rats at a dose as small as 100 mg/kg, whereas, with ip administration at such a small dose, fasting caused no significant alteration in the AUC (toxicity). When rats were exposed by inhalation to CHCl3 vapor, food deprivation had little or no effect on either the blood concentration or the toxicity until the exposure concentration was raised to 500 ppm. The present study indicates that po administration is different from both ip and inh administration with regard to the effect of enzyme induction on the toxicokinetics of CHCl3, mainly due to the first-pass metabolism unique to po administration. PMID- 7482532 TI - Lindane-induced elimination of gap junctional communication in rat uterine myocytes is mediated by an arachidonic acid-sensitive cAMP-independent mechanism. AB - Previous studies by this laboratory showed that the pesticide lindane rapidly and potently inhibits gap junctional communication in myometrial smooth muscle cells. This study examined the possible role of cAMP or arachidonic acid in lindane's elimination of myometrial gap junctional communication. Lindane produced concentration-dependent increases in cAMP of 1.21, 2.94, 6.06, and 8.69 pmol/mg protein with 0.1, 1, 30, and 100 microM lindane, respectively, compared to solvent-treated controls (1.27 pmol/mg protein). Lindane also increased release of tritiated arachidonic acid to 342, 509, 852, 1236, 1639, and 4454 dpm/micrograms protein with 0.01, 0.1, 1, 10, and 100 microM lindane, respectively, compared to solvent controls (342 dpm/micrograms protein). Transfer of Lucifer Yellow dye was used as a measure of gap junctional communication. Both 8-br-cAMP (98, 97, 54, and 4% transfer seen with 0, 1, 10, and 100 microM cAMP) and arachidonic acid (98, 73, 54, 31, and 0% dye transfer for 0.1, 1, 10, 100, and 1000 nM arachidonic acid) depressed dye transfer in cultured myocytes. Although the adenylate cyclase inhibitor 2',3'-dideoxyadenosine completely reversed forskolin-induced depression of dye transfer (1 microM forskolin, 22% transfer), it had no effect with lindane, indicating that lindane's depression of dye transfer was independent of adenylate cyclase activation. Lindane's inhibition of dye transfer was effectively reversed by growing myometrial cells under arachidonic acid-free conditions in the presence of eicosapentaenoic acid, a fatty acid that competes with arachidonic acid for the sn-1,2 position of membrane phospholipids: 0, 15, 40, and 88% dye transfer occurred in the presence of 0.01, 0.1, 1, and 10 microM eicosapentaenoic acid with 30 microM lindane. This implies that arachidonic acid release may be a critical event associated with lindane's inhibition of gap junctional communication in uterine myocytes. PMID- 7482534 TI - Dde chlorinated insecticide 1,1-dichloro-2,2-bis(4-chlorophenyl)ethane (p,p'-DDD) increases intracellular calcium in rat myometrial smooth muscle cells. AB - A previous report from this laboratory showed that two DDT isomers, o,p'-DDT and p,p'-DDD, increased the frequency of spontaneous oscillatory contractions to a similar extent in isolated rat uterine segments. Because regulation of intracellular calcium is fundamental for the development of oscillatory contractions, the present study examined the effects of p,p'-DDD on intracellular free calcium concentration ([Ca2+]i) in individual rat myometrial smooth muscle cells loaded with the fluorescent Ca2+ indicator fura 2. In the presence of extracellular calcium, 50 and 100 microM p,p'-DDD significantly increased peak [Ca2+]i 586 and 921%, respectively, over basal [Ca2+]i. No significant effect was observed with 10 microM p,p'-DDD. In the absence of extracellular calcium, the response to 100 microM p,p'-DDD was significantly attenuated, with cells averaging a 108% increase in peak [Ca2+]i over basal levels, presumably through Ca2+ release from intracellular stores. Nifedipine and cadmium chloride, blockers of voltage-dependent calcium channels, inhibited 100 microM p,p'-DDD-stimulated increases in [Ca2+]i such that peak [Ca2+]i was increased 250% and 259%, respectively. Because of the prominent inhibition observed with the voltage dependent calcium channel blockers, the effect of p,p'-DDD on membrane depolarization was examined using a cationic fluorescent indicator of membrane potential, [diS-C2(5)]. A concentration of 50 microM p,p'-DDD depolarized the cells by 35% of maximum during treatment with p,p'-DDD. The data demonstrate that p,p'-DDD increased [Ca2+]i in rat myometrial smooth muscle cells in a concentration-related manner, and that this increase was largely dependent on influx of extracellular calcium through dihydropyridine-sensitive, voltage dependent calcium channels. The data further show that p,p'-DDD depolarized the plasma membrane, providing a possible mechanism for activation of voltage dependent calcium channels. Additionally, another calcium source, perhaps an intracellular pool, contributes significantly less to the rise of [Ca2+]i. Whether p,p'-DDD initiates the calcium response by direct actions on the plasma membrane or by other means remains to be determined. PMID- 7482533 TI - Constitutive expression of metallothionein-III (MT-III), but not MT-I, inhibits growth when cells become zinc deficient. AB - BHK cells were stably transformed with plasmid constructs that allowed constitutive expression of either mouse metallothionein-I (MT-I) or MT-III to determine whether these isoforms have different physiological properties. Cells expressing equivalent amounts of MT-I or MT-III could grow in 60-fold more cadmium than nontransfected cells and they were 2- to 3-fold more resistant to zinc, copper, and cobalt. The results suggest that the two MT isoforms detoxify these metals similarly. MT-III reduced the amount of zinc available to activate a zinc-sensitive reporter gene; MT-I actually increased the expression of the reporter gene at low concentrations of zinc. Cells expressing MT-I or MT-III also responded differently to zinc deficiency. When cells expressing MT-I were deprived of zinc, the amount of MT-I protein declined to undetectable levels, even though MT-I mRNA was still abundant, and cell proliferation was unaffected. In contrast, when cells expressing the MT-III gene were deprived of zinc, cell proliferation was arrested and MT-III protein persisted. These results suggest that MT-I does not compete with essential zinc-requiring proteins and is degraded, whereas MT-III competes for zinc and exacerbates zinc deficiency. PMID- 7482535 TI - Lysine adducts between methyltetrahydrophthalic anhydride and collagen in guinea pig lung. AB - The formation of adducts between methyltetrahydrophthalic anhydride (MTHPA), an important industrial chemical and potent allergen, and collagen from guinea pig lung tissue was investigated. Collagen peptides were obtained from the lung tissue by homogenization, defatting, washing, and digestion with collagenase. In experiments in vitro, lung tissue was exposed to 8.4 mumol (50 microCi) of 14C MTHPA. The amount of adducts was 97 nmol MTHPA/g of wet tissue as determined from the bound radioactivity. In a study in vivo, four guinea pigs were injected intratracheally with 8.4 mumol of 14C MTHPA each. The amount of adducts was 0-1.2 nmol MTHPA/g of wet tissue (determined by bound radioactivity). N epsilon methyltetrahydrophthaloyl-L-lysine (MTHPL) was synthesized and characterized by NMR, UV, and mass spectrometry (MS). A method to analyze MTHPL, after derivatization with methanol and pentafluorobenzoyl chloride, using gas chromatography-MS was developed. Analysis of Pronase-digested MTHPA-exposed lung tissue showed a concentration of 19 nmol MTHPL/g wet lung in vitro and between 0 and 0.15 nmol MTHPL/g wet lung in vivo. Thus, 20% in vitro and 12-15% in vivo of the bound radioactivity was found as adducts with lysine. These results are a first step toward studies of allergenic epitopes in proteins and methods for biological monitoring of exposure to acid anhydrides. PMID- 7482536 TI - The effect of selenium compounds (selenite, selenate, ebselen) on the production of thromboxane and prostacyclin by the human term placenta in vitro. AB - Selenium not only has an important role in controlling lipid hydroperoxides through glutathione peroxidase (GPX) activity, but also can produce oxidative stress through exposure to selenite. Because levels of lipid hydroperoxides affect the production of thromboxane A2 (TxA2) and prostacyclin (PGI2), selenium compounds may be able to influence the production of these two vasoactive substances. Late-gestation pregnancy reduces the half-life of PGI2; therefore, pregnancy itself may enhance susceptibility to changes in the production of TxA2 and PGI2. The objective of this investigation was to determine if different selenium compounds, selenite, selenate, and ebselen, can influence the human term placental production of thromboxane B2 (TxB2) and 6-keto-prostaglandin F1 alpha (6-keto-PGF1 alpha), the inactive hydrolysis products of TxA2 and PGI2. Although selenate exposure (40 microM 24 hr) increased TxB2 production, and ebselen (an organic selenium compound with GPX activity) exposure (40 microM, 24 hr) decreased both TxB2 and 6-keto-PGF1 alpha production, only selenite had a significant effect on the TxB2/6-keto-PGF1 alpha ratio. Three exposures to selenite at 6 microM (32 hr) significantly decreased 6-keto-PGF1 alpha production with no effect on TxB2 production or tissue GPX activity. Following two exposures to selenite at 20 and 40 microM (24 hr), TxB2 production was significantly increased, while tissue 6-keto-PGF1 alpha production and tissue GPX activity were significantly decreased. These results indicate that selenite, but not selenate or ebselen, can directly affect the human placenta by producing changes in the TxB2/6-keto-PGF1 alpha ratio, which may be related to increased vasoconstriction and blood coagulation. PMID- 7482537 TI - Toxicokinetics of topical sulfur mustard penetration, disposition, and vascular toxicity in isolated perfused porcine skin. AB - Sulfur mustard bis(2-chloroethyl) sulfide (HD) is a bifunctional alkylating agent that causes cutaneous vesication. The isolated perfused porcine skin flap is an in vitro model that has been used to study this toxic response. The purpose of this study was to formulate a toxicokinetic model of HD penetration and cutaneous disposition as an aid in correlating critical steps in the pathogenesis of vesication to HD concentrations in different regions of skin. [14C]HD was dosed topically in ethanol at 10.0 mg/ml in a 7.5-cm2 dosing site and venous efflux samples were collected over 2, 4, or 8 hr. At the termination of the experiment, stratum corneum tape strips, core biopsies for serial sections, and the entire skin flap were collected for radioassay. Peak 14C-radiolabel flux occurred within 5 to 60 min in all skin flaps, much earlier than signs of HD-induced toxicity. A toxicokinetic model was used to quantitate the time profile of HD disposition in different skin compartments. Estimates of vascular and extracellular volume changes due to topical HD toxicity were estimated using radiolabeled albumin and inulin infusions. A second toxicokinetic model, with a time-variant distribution rate, was used to simulate volume changes. In order to accurately predict HD disposition, it was necessary to add another compartment as a reservoir for slowly released metabolites of HD. This model provides a quantitative profile of the time course of HD (or metabolites) disposition within skin which would aid in the interpretation of mechanistic studies of vesication as well as in designing interventive antivesicant drug strategies. PMID- 7482538 TI - Acrolein-induced smooth muscle hyperresponsiveness and eicosanoid release in excised ferret tracheae. AB - Acrolein is a ubiquitous toxic air pollutant that can have adverse lung effects. To understand the mechanism governing airway reactivity in relation to acrolein uptake, in vitro experiments were conducted in which excised tracheae from ferrets were exposed for 1 hr to a unidirectional constant flow (100 ml/min) of an acrolein-in-air mixture at several concentrations (0-12.5 ppm). During exposure, acrolein uptake into the trachea was determined by a chromatographic analysis of gas samples taken at the entrance and at the exit of the trachea. Smooth muscle contractility in response to carbachol (CCh), acetylcholine (ACh), and potassium chloride (KCl) was measured following exposure, and eicosanoids released in the perfusate baths were assayed. The results indicate that the fractional uptake into an excised ferret trachea was strongly dependent on inlet concentration, implying that diffusion and reaction processes of acrolein in airway tissue are not linear. Only the low concentration of acrolein caused an increase of eicosanoid release from the exposed tracheae in the perfusate bath; it is possible that, at higher exposure concentration, the epithelium was sloughed off and most of the eicosanoids were lost. Although acrolein did not alter smooth muscle response to KCl, it did increase the contractile responses to CCh and ACh, suggesting an alteration in the pharmacomechanical but not the electromechanical coupling of ferret tracheal smooth muscle; therefore, it is more likely that this hyperresponsiveness occurs primarily by a mobilization of intracellular Ca2+ stores rather than by an increased influx of extracellular Ca2+ through voltage-dependent channels. PMID- 7482539 TI - Induction of phase I and phase II drug-metabolizing enzyme mRNA, protein, and activity by BHA, ethoxyquin, and oltipraz. AB - Various natural and synthetic compounds are known to protect against cancer by elevating phase II detoxification enzymes. Generally classified as monofunctional, these inducers are believed to trigger cellular signal(s) that activate gene transcription through an antioxidant or electrophile response element (ARE/EpRE) in responsive genes. In contrast, the phase I enzymes of drug metabolism (cytochrome P450s) are not believed to be induced by monofunctional inducers and P450 genes have not been found to contain functional ARE/EpREs. In this study, rats were treated with the monofunctional inducers tert-butylated hydroxyanisole, ethoxyquin, and oltipraz to study the inducibility of individual glutathione S-transferase isozymes, NADP(H):quinone oxidoreductase, gamma glutamylcysteine synthetase, UDP-glucuronosyl transferase, and cytochrome P450 enzymes. Hepatic mRNAs were analyzed on Northern blots using gene-specific oligonucleotide probes for GST Ya1, Ya2, Yc1, Yc2, Yb1, Yb2, and Yf, for UGT 1*06, and for P450 1A1, 1A2, 2B1, 2C11, 3A2, and 4A1. NADP(H):quinone oxidoreductase and gamma-glutamylcysteine synthetase mRNAs were detected using cDNA probes. All the phase II detoxification enzymes analyzed, except GST Yf, were induced by the three monofunctional inducers, suggesting that these genes may be regulated by a mechanism involving an ARE/EpRE element in their promoter region. Interestingly, it was found that ethoxyquin was a particularly good inducer for both members of the P450 2B family, 2B1 and 2B2, and both ethoxyquin and oltipraz were also capable of modestly inducing P450 1A2 and 3A2. Oltipraz was found to slightly induce P450 2B2, but not 2B1, at the dose and time analyzed. Induction of mRNA generally correlated well with induction of protein levels determined by Western blot and/or enzyme activity measurements for selected enzymes. The results of this study suggest that many phase II enzymes may contain ARE/EpRE elements in addition to those confirmed to be regulated by a mechanism involving ARE/EpRE elements. In addition, it was found that several P450 enzymes were induced by monofunctional inducers, suggesting a possibility that some phase I enzymes may also be regulated by a mechanism involving ARE/EpRE elements. PMID- 7482541 TI - Effects of developmental hypothyroidism on auditory and motor function in the rat. AB - Deafness is a common result of severe hypothyroidism during development in humans and laboratory animals; however, little is known regarding the sensitivity of the auditory system to more moderate changes in thyroid hormone homeostasis. The current investigation compared the relative sensitivity of auditory function, motor function, and growth to the effects of moderate to severe perinatal hypothyroidism in the rat. Rats received propylthiouracil (PTU) in drinking water at concentrations of 0, 1, 5, and 25 ppm from Gestation Day 18 until postnatal day (PND) 21, and the effects on their offspring were evaluated. At 1 ppm, PTU did not affect any of the measured endpoints. Serum thyroxin concentrations were sharply reduced in the 5 and 25 ppm PTU groups at all ages sampled (PND 1, 7, 14, and 21). Marked reductions in serum triiodothyronine (T3) concentrations were also detected for all ages > or = 7 at 25 ppm PTU, whereas no effects of 5 ppm PTU on serum T3 were apparent until PND 21. Compared to the controls, pups exposed to the highest dose of PTU demonstrated a delay in eye opening, reduced body weights, decreased and/or delayed preweaning motor activity, and persistent, postweaning hyperactivity. Only slight and transient effects on eye opening and ontogeny of motor activity were seen at the intermediate dose of PTU (5 ppm). Reflex modification audiometry revealed that, compared to controls, adult offspring from the 5 and 25 ppm treatment groups showed dose-dependent auditory threshold deficits (35 to > 50 dB) at all frequencies tested (1, 4, 16, 32, and 40 kHz). Such dose-dependent effects indicate that the developing auditory system may be sensitive to mild hypothyroidism, suggesting the possible need for routine audiometric screening for infants and children at risk for iodine deficiency, myxedema, and/or exposure to thyrotoxic environmental agents. PMID- 7482540 TI - Axonal atrophy is a specific component of 2,5-hexanedione peripheral neuropathy. AB - To assess the relevance of previously identified axonal atrophy to hexanedione neuropathy, the present study quantitated fiber size in peripheral nerve of rats intoxicated with 2,5-hexanedione (HD) by either oral ingestion (0.4% in drinking water) or ip injection (0.4 g/kg/day). Prior to the appearance of neurobehavioral deficits, rats exposed to oral HD (77 days) exhibited axonal atrophy in proximal sciatic nerve and giant axonal swellings in distal tibial nerve. As oral-treated rats progressed to moderate (86 days) and severe (103 days) hindlimb weakness, both nerve regions contained a mixed population of atrophied and swollen axons. Rats injected with HD ip were sampled at behavioral endpoints that matched those of oral HD-treated rats. In sciatic and tibial nerves from rats treated ip, reductions in the axon area were similar to oral exposure. However, ip treatment did not produce giant axonal swellings in either nerve. Thus, although both routes of administration caused equivalent behavioral neurotoxicity, the expression of axonal swellings was route-dependent. This suggests that the production of swellings depends upon the HD exposure pattern. In contrast, axonal atrophy was prevalent in both nerve regions sampled and developed in parallel with behavioral deficits. In addition, atrophy was expressed regardless of the intoxication route which indicates that atrophy can occur independent of axonal swellings. Together, these attributes suggest that atrophy is a specific component of HD neurotoxicity. PMID- 7482542 TI - Developmental exposure to polychlorinated biphenyls (Aroclor 1254) reduces circulating thyroid hormone concentrations and causes hearing deficits in rats. AB - Developmental hypothyroidism causes growth deficits, motor dysfunction, and hearing disorders in humans and animals. Therefore, environmental toxicants, such as polychlorinated biphenyls (PCBs), may secondarily affect these endpoints via thyrotoxicity. In this study, Long-Evans rats were given Aroclor 1254 (po), at 0, 1, 4, or 8 mg/kg from Gestation Day 6 through Postnatal Day (PND) 21. We evaluated the offspring at various age intervals for circulating thyroid hormone concentrations [thyroid-stimulating hormone, and free and total triiodothyronine (T3) and thyroxin (T4)], body weight, eye opening, survival, motor activity development, auditory startle response, and auditory thresholds. Circulating T4 concentrations were sharply reduced in a dose-dependent fashion in PCB-exposed groups at PND 1, 7, 14, 21, and 30 but recovered to control levels by PND 45. Moderate reductions in T3 concentrations were apparent in the 4 and 8 mg/kg groups on PND 21 and 30. Deficits in body weight gain and early eye opening were apparent in the treated pups; by weaning, pup mortality was 20% in the 4 mg/kg group and 50% at the highest dose. Motor activity was also transiently reduced in 15 day old offspring from the 8 mg/kg group. At this dose, animals showed reduced auditory startle amplitudes at PND 24, but not when tested as adults. Importantly, Aroclor 1254 caused permanent auditory deficits (20-30 dB threshold shift) at the lowest frequency tested (1 kHz) in both the 4 and 8 mg/kg groups, whereas auditory thresholds were not significantly affected at higher frequencies (4, 16, 32, or 40 kHz). These data indicate that while some effects of Aroclor 1254 exposure are dissimilar to drug-induced hypothyroidism (e.g., age of eye opening), effects on hormone levels and body weight are comparable. Detection of auditory deficits in PCB-treated animals is a novel finding and may reflect the effects of thyroid hormone disruption on the development of the cochlea. PMID- 7482543 TI - Expression and inducibility of antigens in severe combined immunodeficient mice recognized by human anti-P450 antibodies. AB - Engrafting components of human immune systems in severe combined immunodeficient (SCID) mice has been utilized to investigate the pathogenesis of several human autoimmune diseases and may provide a model for studying idiosyncratic drug toxicity. The purpose of this investigation was to examine in SCID mice the tissue distribution and inducibility of antigens recognized by anti-cytochrome P450 (CYP) antibodies in sera from patients with hypersensitivity reactions to the aromatic anticonvulsants phenytoin, phenobarbital, and carbamazepine. Microsomal proteins were prepared from liver, skin, kidney, intestine, and lung of SCID mice pretreated with vehicle (50% propylene glycol/DMSO), phenytoin, carbamazepine, phenobarbital, or dexamethasone. Proteins immunoreactive with anti CYP2C and anti-CYP3A antisera were detected in all organs examined. Antibodies in patient sera recognized a 53-kDa hepatic microsomal protein that was expressed to a limited extent in vehicle-pretreated microsomes, but which was induced by dexamethasone, phenytoin, and phenobarbital, but not carbamazepine. This antigen was very similar to a 52.5-kDa protein immunoreactive with anti-CYP3A polyclonal antibody. The expression and inducibility of the 53-kDa antigen correlated significantly with testosterone 6 beta-, 2 beta-, and 15 beta-hydroxylation and erythromycin N-demethylase activity, all markers of CYP3A activity, and is tentatively identified as CYP3A11. No immunoreactivity was observed in murine extrahepatic organs including skin, kidney, intestine, and lung. Therefore, further development of the SCID mouse model may require xenotransplantation of human target organs like skin together with transfer of patient immune systems to reproduce the serological and pathological features of human anticonvulsant hypersensitivity reactions. PMID- 7482544 TI - Conjugation of acrylonitrile and 2-cyanoethylene oxide with hepatic glutathione. AB - The glutathione (GSH) conjugation of the rat carcinogen acrylonitrile (ACN) and its epoxide metabolite 2-cyanoethylene oxide (CEO) by rat, mouse, and human liver enzymes was characterized in vitro since GSH conjugation is the major disposition pathway for these chemicals in vivo. Mass spectral analyses indicated that S-(2 cyanoethyl)GSH was the product from reaction of GSH and ACN and that S (cyanohydroxyethyl)GSH reaction products were formed from CEO. Because of the rapid nonenzymic reactions of ACN and CEO with GSH at pH 7.3, the steady-state kinetics of hepatic GSH conjugation were determined at pH 6.5 by HPLC analysis of the products. Hyperbolic kinetics were observed with respect to GSH for the reactions catalyzed by mouse or rat hepatic cytosols at pH 6.5, whereas sigmoidal kinetics were observed with respect to ACN or CEO. This kinetic pattern is consistent with the random sequential kinetic mechanism that has been described for GSH S-transferases. Estimates of the maximal velocities of the reaction at pH 6.5 showed that mouse enzymes had a 4- to 6-fold greater capacity for GSH conjugation of ACN and CEO than rat enzymes. ACN appeared to be conjugated with GSH more efficiently than CEO under these conditions. At physiological pH (7.3), rapid nonenzymic conjugation of GSH (10 mM) with ACN or CEO (5 mM) was observed (approximately 25 and 15 nmol product/min, respectively). Addition of hepatic cytosols or microsomes from rats or mice increased the velocity of GSH conjugation approximately 1.6-fold. A similar velocity enhancement was observed with human liver cytosols for the GSH conjugation of ACN, but not for CEO. Human liver microsomes did not enhance the velocity of GSH conjugation of either substrate. These results suggest that ACN is a better substrate for human liver GSH S-transferases than CEO. Estimation of the initial velocities of the GSH conjugation reactions in intact rodent liver from the in vitro data at pH 7.3 suggests that the enzyme-mediated GSH conjugation of ACN and CEO will be approximately 4-fold greater than the velocity of the direct chemical reaction with GSH. PMID- 7482545 TI - Biological Mechanisms and Quantitative Risk Assessment. Proceedings of a symposium. November 1-4, 1993. PMID- 7482546 TI - Dioxins: model chemicals for assessing receptor-mediated toxicity. AB - Dioxins and related compounds are chlorinated aromatic hydrocarbons that are persistent in both environmental and biological samples. Many members of this class of compounds produce a similar spectrum of toxicity which is mediated by interaction with the Ah receptor. The toxic effects of these chemicals can best be described by their actions as growth dysregulators. Dioxins disrupt normal homeostatic processes that tightly regulate cellular growth and differentiation. Disruption in these processes produce a variety of toxicities and pathologies. The available data indicate that humans are sensitive to the toxic effects of these chemicals. Clearer definition of human responses and the body burdens associated with such effects requires more research. Comprehensive risk assessments of dioxins should include all Ah receptor ligands such as the halogenated dibenzofurans and biphenyls. PMID- 7482547 TI - Biologically-based models of dioxin pharmacokinetics. AB - Significant advances have been made in the development of physiologically-based models of dioxin pharmacokinetics (PBPK) in the last 5-6 years. These models incorporate explicit descriptions of biological factors which determine tissue dosimetry of dioxin and include some description of dioxin-mediated pharmacodynamic events. Biological determinants of dioxin disposition include fat solubility, specific and inducible binding in the liver, diffusion-limited tissue distribution and metabolic elimination. PBPK models have been successfully used to predict the dose and time-dependent chemical disposition and protein induction properties of 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD) over a wide variety of experimental data sets with rodents. The models have also been extended to describe the disposition of a brominated dioxin, 2,3,7,8-tetrabromodibenzo-p dioxin. As these quantitative descriptions of disposition are more fully refined, particularly with regard to pharmacodynamic descriptions of dioxin-mediated alterations in gene expression, more accurate predictions of tissue dosimetry and tissue responses will be performed across dose, species and related polyhalogenated aromatic hydrocarbons. Accurate, mechanistic dosimetry models will facilitate biologically-based approaches to the human risk assessment of these important and ubiquitous environmental contaminants. PMID- 7482548 TI - Biochemical mechanisms and cancer risk assessment models for dioxin. AB - Biologically realistic mechanistic models of carcinogenesis by TCDD are composed of equations representing biochemical events leading to altered expression of proteins involved in the response or equations representing the kinetics of proliferation of clones of mutant cells. A biochemically augmented physiological dosimetry model reproduces the observed altered expression of liver proteins in female rats exposed to dioxin. The model suggests that oxidation of estradiol to DNA reactive quinones or semiquinones by CYP1A2 protein induced by TCDD may contribute to an increased mutational rate. It suggests that TCDD-stimulated production of a peptide ligand of the epidermal growth factor (EGF) receptor and subsequent activation of the receptor's tyrosine kinase activity may increase the rate of proliferation of susceptible cells. These calculated quantities can serve as indices of toxicity and can be used to predict tumor incidence as a function of exposure. PMID- 7482550 TI - Cell proliferation in the bladder and implications for cancer risk assessment. AB - Chemicals can increase carcinogenic risk by either directly damaging DNA or increasing cell replication or they can do both. These effects have different implications for a biologically-based extrapolation from rodent bioassays to humans. 2-Acetylaminofluorene (2-AAF) administered at low doses to mice for a lifetime has a different dose-response for the liver (approximately linear) compared to the urinary bladder (apparent no effect dose of 45 ppm with a sigmoidal dose response at 60-150 ppm), which can be explained if carcinogen metabolism, DNA adduct formation and cell proliferation effects are considered. In contrast to 2-AAF and other genotoxic chemicals, chemicals which form calculi in the urine do not generally damage DNA directly but increase cell proliferation dramatically by eroding the bladder surface, leading to regenerative hyperplasia. This occurs only at doses at which calculi form; lower doses do not produce calculi and, therefore, do not increase cell proliferation or cause tumors. Extrapolation to humans from the rodent bioassay should be dependent on dose requirements for formation of calculi rather than any type of statistical extrapolation to lower doses. Saccharin and other sodium salts administered at high doses to rats also produce bladder cancer by increasing cell proliferation. These salts do not affect mice, hamsters, guinea pig or monkeys. Based on dose and mechanistic considerations, saccharin and these other sodium salts are unlikely to be human carcinogens. Extrapolation to possible human cancer risk requires biological determinations rather than simply using statistical formulations. PMID- 7482549 TI - Incorporating cell proliferation kinetics into models for cancer risk assessment. AB - Some general principles in incorporating cell proliferation kinetics into dose response models for cancer risk assessment are discussed. Two examples are presented in which a biologically-based dose-response model explicitly incorporating cell proliferation kinetics was used for the analysis of toxicologic data. In the first example, analysis of an initiation-promotion experiment in the rat liver, with diethylnitrosamine (DEN) as initiator and with 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD) and 1,2,3,4,6,7,8-heptachlorodibenzo-p dioxin (HCDD) as promoters, is presented. The results of the analysis indicate that, in addition to its promoting activity, TCDD has weak initiating activity. In the second example, multiple doses of N-nitrosomorpholine (NNM) were administered to rats in their drinking water and quantitative information on ATPase deficient foci in the liver recorded at various times. Additionally, a separate group of animals, administered the same doses, was followed until death and the presence or absence of malignant liver tumors recorded. The parameters of the model were estimated by fitting the model to these data and the estimated parameters were used to construct a dose-response curve for the probability of malignant tumors. Few malignant tumors were observed at the lower doses; however, the information on ATPase deficient foci can be used in the model to extend the range of the dose-response curve below the doses at which malignant tumors are observed. PMID- 7482551 TI - Comparison of experimental and theoretical parameters of the Moolgavkar-Venzon Knudson incidence function for the stages of initiation and promotion in rat hepatocarcinogenesis. AB - Mathematical descriptions of complex biological phenomena, such as cancer, require an experimental format that faithfully recapitulates the biological process. In addition, the biological process must dictate the parameters in the mathematical formula. Evidence from the epidemiology of several human cancers and from experimental carcinogenesis in several organ systems indicates that cancer is a multistage process. The initiation-promotion-progression format of experimental carcinogenesis mimics the development of cancer in humans and other animals. In rats, the altered hepatic focus model of hepatocarcinogenesis has been well characterized and, coupled with the method of quantitative stereology, permits accurate determination of the number and the volume fraction of such altered foci per liver. The placental isozyme of glutathione S-transferase (PGST) is reportedly the best single marker of preneoplasia in the rat liver. Recently, single hepatocytes expressing PGST have been proposed as putatively initiated cells. Quantitation of individual hepatic cells and altered hepatic foci expressing PGST in the livers of rats subjected to an initiation-promotion protocol permits determination of the congruence of the Moolgavkar-Venzon-Knudson (MVK) model with experimental data. The best fit of the MVK model for the preneoplastic stages of hepatocarcinogenesis assumes that all hepatocytes are susceptible and that single hepatocytes expressing PGST are the initiated cell population for the focal lesions that express PGST. Further refinement of the initiation-promotion-progression model to permit accurate quantitation of early malignant conversion should allow a more complete analysis of the congruence of the MVK model for human cancer risk determination. In addition, the MVK model may be extended to other model systems and to human cancers in which early preneoplasia can be quantitated. Furthermore, the use of a more biologically based risk-assessment protocol, such as the MVK model rather than the stochastic one-hit model presently used, would permit incorporation of the present knowledge on the pathogenesis of cancer. To apply experimental data to a mathematical model that reflects the biological processes underlying human cancer development will require integration of the cell kinetics and experimental data to a mathematical model that reflects the biological processes underlying human cancer development including the pharmacokinetic and pharmacodynamic properties of the treatment chemicals. PMID- 7482552 TI - Cancer and non-cancer risk assessment: not so different if you consider mechanisms. AB - Default risk assessment procedures use threshold models for non-carcinogens and a non-threshold model for carcinogens. This a priori distinction reflects the fact that the default procedures do not consider mechanisms of action of specific chemicals. When mechanisms are considered, the distinction is not necessary. Starting with the premise that the goal of risk assessment is to identify actual risk for specific chemicals, three major, generic components of the overall mechanism translating exposure into a response of regulatory interest are identified. These are the specific mechanisms linking (1) exposure with dose to target tissue, (2) target tissue dose with short-term responses such as cytolethality or mutation, and (3) short-term responses with ensuing long-term responses such as cancer or cirrhosis. (Short-term responses may be regulatory end points of interest, or they may be intermediate steps on the way to longer term sequelae). On-going research on formaldehyde and chloroform is described to illustrate how these three components of the overall mechanism can be examined experimentally and used in specific models. The impact of mechanism-based risk assessment on uncertainty is also considered. Uncertainty is a function of the extent to which the model used for risk assessment misspecifies the actual mechanism of action for the chemical in question. There is a trade-off between (a) mechanism-based models that may reduce uncertainty but are expensive and time consuming to develop and (b) default models that are not chemical-specific but can be used with minimal data sets. Experience with mechanism-based risk assessment may allow modification of default procedures to minimize this trade off. A future default procedure for carcinogen risk assessment might allow specification of mode of action. For example, while DNA reactive-carcinogens would still be assumed to have linear low-dose risk, carcinogens acting through purely cytotoxic mechanisms might be assumed to have sharply non-linear or even threshold dose-response curves. PMID- 7482553 TI - Pharmacokinetics and receptor-mediated pharmacodynamics of corticosteroids. AB - The corticosteroids, such as prednisolone and methylprednisolone, provide diverse antiinflammatory and immunosuppressive effects which typically show responses with slow onset and prolonged duration. This report summarizes modeling efforts which are successful in describing such steroid effects. Clinical effects with such a pattern, including adrenal suppression and altered trafficking of basophils and helper T-cells, can be related to plasma drug concentrations by models containing an inhibition function and differential equations for controlling input and disposition of the response variable. Some responses have circadian-controlled inputs which add time-dependent complexities to the models. Kinetic/dynamic data for several corticosteroid effects yield IC50 values which agree well with receptor KD values. A relationship of linear AUC of effect versus log AUC of steroid in plasma is found with these models over a large range of doses. Gene-mediated effects of corticosteroids are initiated by receptor-binding which causes a cascade effect altering DNA transcription, RNA, mRNA and proteins or enzymes accounting for drug effects. Models for such behavior have been developed in animals for hepatic tyrosine aminotransferase (TAT) enzyme activity. Studies with methylprednisolone formulated in liposomes show tissue sequestration of steroid, prolonged receptor-binding and extended inhibition of splenocyte proliferation. The data and models usually show good correspondence of the AUC of receptor occupancy with the AUC of pharmacologic response. PMID- 7482554 TI - Toxicity prediction from metabolic pathway modelling. AB - The technique of kinetic simulation, in which a metabolic pathway is modelled by a system of differential equations, has been used to study several questions concerning the teratogenic effects of antifolate drugs. These studies made use of BioCHIMICA, a new biochemical modelling software package. This program is a general-purpose system for simulation of reaction networks. It can simulate systems of over 100 reactions, and can make use of a wide variety of rate equations. Several applications of this program in predicting the teratogenicity of antifolates are described: (a) prediction of relative toxic potency of related analogues, (b) prediction of shapes of dose-response curves in an attempt to determine whether safe exposure levels may exist, (c) prediction of reversibility, i.e. can we distinguish between reversible damage and effects likely to be irreversible, (d) prediction of combined inhibitor effects, i.e. can we identify which teratogenic effects are likely to be synergistic and which antagonistic? These questions were asked in an attempt to predict the toxic effects of a group of experimental inhibitors of thymidylate synthase and dihydrofolate reductase. PMID- 7482556 TI - Regulatory and research issues related to cholinesterase inhibition. AB - Assessing the neurotoxic potential of organophosphate and carbamate pesticides should be greatly facilitated by the knowledge that the mechanism of action of these insecticides is presumed to be the inhibition of cholinesterase, the enzyme which controls the levels of neurotransmitter, acetycholine. Although the inhibition of cholinesterase activity is the recognized mechanism of action, many questions remain regarding the use of cholinesterase inhibition data as a critical effect for establishing risk of cholinesterase-inhibiting pesticides. Specifically, questions have arisen regarding whether blood cholinesterase inhibition correlates with inhibition in target tissues (e.g. brain or muscle) and whether cholinesterase inhibition in any tissue correlates with the adverse clinical and behavioral effects produced by exposure to cholinesterase-inhibiting pesticides. Studies in our laboratory indicate that blood cholinesterase inhibition in both acute and subchronic dosing regimens correlates with inhibition in other tissues, if measurements are taken at the appropriate times. Moreover, there is evidence in the literature and from our laboratory that cholinesterase inhibition correlates with the emergence and severity of clinical signs of poisoning by cholinesterase-inhibiting pesticides. PMID- 7482555 TI - Biological modeling of 5-fluorouracil developmental toxicity. AB - A biologically-based dose-response (BBDR) model is a mathematical description of the biological events leading to expression of a toxic response. As an alternative to current approaches in non-cancer risk assessment, such models will reduce uncertainty in that they will provide a more comprehensive description of toxicity. We are involved in construction of a BBDR model for the developmental toxicity of 5-fluorouracil (5-FU) in the rat using multiple approaches. First, to identify critical events in the pathogenesis of 5-FU developmental toxicity, thymidylate synthetase (TS) inhibition and alterations in cell kinetics and growth were examined in embryos following maternal administration of 5-FU on day 14 of gestation. A dose-related decline in TS activity was observed within 1 h; however, maximal inhibition and recovery were similar at 10, 20 and 40 mg/kg. Dose-dependent cell cycle alterations were observed within 4 h after exposure and were maximal at 8 h. Hindlimb growth reduction was observed 24 h after exposure to 40 mg/kg, but not at lower doses. At term hindlimb defects were observed at doses above 30 mg/kg. An integrated dose-response model for hindlimb defects was derived from empirical relationships among these events. The resultant dose response somewhat over-predicted the developmental toxicity of 5-FU, although results of a Monte Carlo simulation indicated that these data were not incompatible with model predictions. Overall, the results suggest that TS inhibition is a key component of the mechanism of 5-FU developmental toxicology, but the model does not capture all of the critical events in the induction of hindlimb defects. A preliminary mechanistic model for the inhibition of embryonic TS, DNA synthesis and cell cycle following maternal exposure to 5-FU, independently derived from literature data to further examine the potential role of this pathway in its developmental toxicity, predicted a dose-response for TS inhibition and DNA synthesis that closely reflected the observed patterns. These results further suggest that TS inhibition, resultant deficits in DNA synthesis and cell cycle perturbations represent a critical mechanistic pathway in the developmental toxicity of 5-FU. PMID- 7482558 TI - Assessing the risk assessment paradigm. AB - Risk assessment provides an essential framework for organizing, evaluating and characterizing scientific information on the nature and magnitude of hazards from specific chemicals. Such characterization of risk is needed for risk communication and for risk reduction and management. Each element of the risk assessment paradigm--hazard identification, dose-response, exposure analysis and risk characterization--is the target of important efforts to improve methodologies and the biological plausibility and clinical significance of our conclusions. PMID- 7482557 TI - Negative selection in hepatic tumor promotion in relation to cancer risk assessment. AB - Mechanistic studies with phenobarbital (PB), 2,3,7,8,-tetrachlorodibenzo-p-dioxin (TCDD) and other liver tumor promoters support a general model of promotion involving negative selection where specifically-mutated cells derive a growth advantage in the presence of persistent mitosuppression. Exposure to these liver tumor promoters appears to transiently enhance hepatocyte replication, presumably via transcriptional activation of growth regulatory genes, leading to a homeostatic increase in mitoinhibitory growth factors in the liver to constrain proliferation. Transforming growth factor beta 1 (TGF-beta), a potent mitoinhibitory growth factor for hepatocytes, has been associated with the mitosuppression caused by PB and certain peroxisomal proliferators. Escape from TGF-beta mitosuppression may involve loss or alteration of function of the mannose 6-phosphate/insulin-like growth factor II (M6P/IGFII) receptor, which is required for TGF-beta 1 activation, or alterations of the TGF-beta types I, II and III signal transduction receptors. A risk assessment based on a negative selection mechanism could be conducted for tumor promotion endpoints with TCDD and compared with current approaches that implicitly regard TCDD as an initiator. Benchmark dose calculation using centrilobular induction of cytochromes P450 1A1 and 1A2 as a surrogate for periportal growth stimulation would provide a rational starting point for application of conventional safety factor approaches, similar to those used with non-cancer effects. In the future, tissue and plasma concentrations of specific growth factors, e.g. TGF-beta or hepatocyte growth factor, HGF, might be considered as more direct dose surrogates for tumor promoting effects of xenobiotics. Uncertainty factor adjustments to a TCDD benchmark dose calculation should eventually rely on direct knowledge of regulation of specific growth regulatory genes and their receptors in relevant species and on species differences in TCDD pharmacokinetics, instead of application of default animal-to-human and interindividual uncertainty factors. PMID- 7482559 TI - Risk assessment and biological mechanisms: lessons learned, future opportunities. AB - During the past decade, toxicological research has been dominated by two themes; investigations to elucidate the mechanisms of action of toxicants and studies to provide information to support improved assessments of human health risks. The conduct of mechanistic investigations was given an early impetus by advances in biochemistry and cell biology and, more recently, by related advances in molecular biology. Research to provide information for improved human health risk assessments was stimulated by the 1983 NAS/NRC report that provided a codified structure for conducting risk assessments. At first glance, it would appear that the two themes are closely related and, indeed, should represent parts of a joined theme. However, examination of the toxicology/risk assessment literature of the past decade indicates that this has not been the case. Reports of mechanistic studies infrequently indicate how the information can be used to provide improved estimates of human risk from exposure to toxicants. If reference is made, it is usually qualitative in nature. Neither is examination of the risk assessment literature reassuring. Mechanistic studies may be cited; however, the final step of the process, risk characterization, is usually dominated by the use of default options grounded in conservative interpretations of generic scientific knowledge. Two examples are reviewed that stand out as illustrations of how mechanistic information can be used to make a difference in risk assessments: (1) consideration of the alpha 2u-globulin-mediated mechanism for evaluating male rat data for relevance in assessing human risks of renal cancer and (2) the use of DNA-protein cross-links as an internal dose metric in cross-species extrapolation of nasal cancer risks from inhaled formaldehyde. This paper reviews past experience on these topics and suggests a strategy for increasing the use of mechanistic information in risk assessments. A key component of the strategy is to use the risk assessment process to identify research needs/opportunities that, if addressed, will reduce the use of default options, thereby reducing the uncertainties in risk assessments. Another component of the strategy is to identify a few chemicals anticipated to exert their effect via different mechanisms and whose mechanisms of dosimetry and disease pathogenesis can be investigated in-depth within a risk assessment framework; this identification will create prototype approaches as alternatives to the use of default options that have major impact on the outcome of the risk assessment process. PMID- 7482560 TI - Historical perspective on risk assessment in the federal government. AB - This article traces the evolution of risk assessment as an essential analytical tool in the federal government. In many programs and agencies, decisions cannot be made without the benefit of information from risk assessment. Although this analytical tool influences important public health and economic decisions, there is widespread dissatisfaction with the day-to-day practice of risk assessment. The article describes the sources of dissatisfaction that have been voiced by scientists, regulators, interest groups and ordinary citizens. Problems include the use of arbitrary exposure scenarios, the misuse of the 'carcinogen' label, the excessive reliance on animal cancer tests, the lack of formal uncertainty analysis the low priority assigned to noncancer endpoints, the poor communication of risk estimates and the neglect of inequities in the distribution of risk. Despite these limitations, the article argues that more danger rests in efforts to make decisions without any risk assessment. Recent Congressional and Administration interest in risk assessment is encouraging because it offers promise to learn from past mistakes and set in motion steps to enhance the risk assessment process. PMID- 7482561 TI - Research to strengthen the scientific basis for health risk assessment: a survey of the context and rationale for mechanistically based methods and models. AB - Assessment of health risks is an integral part of regulatory decision-making that occurs at the interface between science (e.g. facts) and policy (e.g. values). Because existing scientific knowledge and understanding are often inadequate to answer the most critical risk-related questions, regulatory agencies have developed sets of formalized 'science policies' to extrapolate from existing data to real-life events and situations. These science policies, as, for example, the use of default assumptions or exposure scenarios, can introduce significant uncertainties into the final risk estimate. We survey the rationale for research to reduce extrapolation-related uncertainties, focusing specifically on the need to develop mechanistically based methods and models, including test methods to identify and characterize health effects, integrated human exposure models, physiologically based pharmacokinetic (PBPK) models and biologically based dose response (BBDR) models. PMID- 7482562 TI - The future of mechanistic research in risk assessment: where are we going and can we get there from here? AB - Quantitative estimates of human health risk are often based on mathematical models fit to experimental or epidemiological data. Recent years have witnessed a trend towards the use of mechanistic models in risk assessment applications. Such models afford a more biologically based interpretation of the data and a firmer scientific basis for extrapolation beyond the conditions under which the original data were obtained. In this article, we review some recent advances in the development of biologically based models for mutagenesis, carcinogenesis and developmental toxicity. Pharmacokinetic and receptor-binding models and their roles in mechanistic risk assessment are also discussed. The future of mechanistic research in risk assessment is contemplated, including the need for more elaborate experiments to obtain the data necessary for mechanistic modeling. PMID- 7482564 TI - Incorporating biological information in quantitative risk assessment: an example with methylene chloride. AB - The interplay between chemical risk assessment and scientific research is discussed in the context of recent attempts to improve the scientific basis for estimates of the human carcinogenic risk from methylene chloride. A combination of basic biochemical research and risk assessment oriented research, both mechanistic and pharmacokinetic, provided the initial impetus for re-evaluating the use of the default risk estimation approach. Resulting efforts to incorporate the new scientific information into the risk assessment process in turn identified specific additional research needed to reduce uncertainty in the estimated risk. This healthy interchange between the two disciplines has served both to refine the human risk estimates for methylene chloride and to more clearly identify key scientific issues for chemical risk assessment in general. PMID- 7482563 TI - Hepatic and pulmonary carcinogenicity of methylene chloride in mice: a search for mechanisms. AB - An inhalation study utilizing over 1400 female B6C3F1 mice was undertaken to study mechanistic factors associated with liver and lung tumor induction following exposure to 2000 ppm of methylene chloride. Mice were exposed to methylene chloride (treated) or chamber air (controls) 6 h per day, for varying durations up to 104 weeks. Several interim sacrifices and 'stop exposures' were included. Exposure to 2000 ppm methylene chloride caused an increase in liver and lung neoplasia in the absence of overt cytotoxicity. Measurement of replicative DNA synthesis done after 13, 26, 52 and 78 weeks of exposure showed a significant decrease in the hepatocyte labeling index at 13 weeks. Replicative DNA synthesis in pulmonary airways after 1, 2, 3, 4, 13 and 26 weeks of exposure to methylene chloride was significantly lower than in air-exposed controls. Likewise, the increase in tumor induction in treated mice was not associated with increased replicative DNA synthesis in liver foci or in alveolar parenchyma. The frequency and pattern of H-ras gene activation were similar in control and methylene chloride-induced liver neoplasms. Similarly, the frequency and pattern of K-ras activation in lung neoplasms were not altered by exposure to methylene chloride. Early exposure to methylene chloride for only 26 weeks was sufficient to cause an increase in lung tumors by 2 years, suggesting that methylene chloride may cause early and persistent loss of growth control in lung cells. This implies that risk management strategies should be aimed at minimizing or eliminating exposure to methylene chloride. Liver neoplasms continued to increase in incidence and multiplicity as exposure continued, suggesting that methylene chloride-induced hepatocarcinogenesis is facilitated by continuing exposure to methylene chloride. Since methylene chloride is a more potent inducer of lung than liver neoplasia, it is recommended that health risk assessment be based on the lung data. While no novel molecular lesions have been found to explain the induction of lung and liver neoplasia in mice, ongoing studies may identify other molecular changes that are important in the genesis of these neoplasms. Hence, it may be necessary to revise risk assessment and management strategies in light of future research findings. PMID- 7482565 TI - Use of quantitative modelling in methylene chloride risk assessment. AB - The benefits of basing quantitative risk assessment on measures of 'internal dose', i.e. target organ exposures as estimated, for instance, by pharmacokinetic models, have been extensively discussed. Recasting risk assessment methods at the level of internal dose raises novel issues, however, some of which are explored by examining the 1987 revision by the US Environmental Protection Agency (EPA) of its cancer risk assessment for inhaled methylene chloride, which was based on the 1987 pharmacokinetic model results of Andersen and coworkers. The internal dose measure was the daily amount of methylene chloride metabolized by a glutathione-S transferase pathway per 1 of target organ (liver and lung). Owing to high-dose saturation of a competing detoxification reaction, this metabolic activation is less-than-proportionally active at low exposure levels. For a given inhalation exposure, humans have relatively less metabolic activation than do mice, but this is shown to be a foreseeable consequence of their relatively lower breathing rate, a cross-species difference already accounted for in standard EPA methodology. Indeed, many species differences in the rates and tempos of physiological processes evince regular 'scaling' relationships across differently sized mammals. EPA's practice of scaling carcinogen doses by body surface area for cross-species extrapolation, often viewed as a correction for metabolic activation, is shown to be more reasonably regarded as an accommodation for the more general species variation in the pace of physiological processes underlying both pharmacokinetics and the carcinogenic response to internal doses. Under this view, the issue of cross-species dose scaling is not obviated by the use of pharmacokinetics. PMID- 7482567 TI - Porphyrin production and excretion by long-term cultures of adult rat hepatocytes and effect of lead exposure. AB - Porphyrin production and excretion and the effects of lead exposure were studied in long-term cultures of adult rat hepatocytes cultured on a feeder layer of 3T3 cells after addition of 5-aminolevulinic acid. Porphyrin excretion into the culture medium showed an irregular profile during the first 10 days, with a maximum increase of 50% at day 4 and at day 10 a value similar to that of day 1. Thereafter, porphyrin excretion decreased progressively to 18% of the initial value after 4 weeks. The cellular porphyrin content, after 7 and 28 days in culture, reached values 3.8 and 2.4-fold higher than the corresponding day 1 value. The exposure to 0.5 and 2.4 microM Pb2+ for up to 28 days produced a biphasic effect on porphyrin excretion. Firstly, there was a progressive decrease up to 81% during the first 6 days of lead exposure and, secondly, this effect was followed by an increase reaching control values at day 15 and of up to 6.7-fold after 22 days of exposure to 2.4 microM Pb2+. Similar changes were observed in cellular porphyrin content. The exposure to 0.5 and 2.4 microM Pb2+ for 2 and 4 weeks also produced morphological alterations and release of cytoplasmic enzymes. Our results show that hepatocytes cultured on 3T3 cells produce and excrete porphyrins for 28 days and that exposure for 4 weeks to micromolar lead concentrations alters these functions and cell morphology and produces cytotoxic effects which are better evaluated by monitoring alterations in porphyrin excretion than by enzyme leakage. They also suggest that this culture system is a useful model for assessing the toxic effects of xenobiotics on the biosynthesis of heme by liver cells. PMID- 7482566 TI - In vitro murine embryotoxicity of cyclophosphamide in embryos co-cultured with maternal hepatocytes: development and application of a murine embryo-hepatocyte co-culture model. AB - The technique of whole embryo culture provides a sensitive model to evaluate both the effects, and their underlying mechanisms, of drugs and environmental chemicals on embryonic development, independent of maternal influences. However, before teratogenic expression, many teratogens must be enzymatically bioactivated to toxic reactive intermediates. To detect such proteratogens, the embryo culture model may need to be coupled with an exogenous bioactivating system if maternal and/or placental metabolism is involved. We developed a similar embryo-hepatocyte co-culture system using embryos and maternal hepatocytes from mice, which often are more sensitive than rats to chemical teratogens, and which may have a balance of phase II drug metabolising enzymes more similar to humans. This murine system was then used to evaluate the relative maternal and embryonic contributions to cyclophosphamide embryopathy. Day 9.5 (morning of plug = day 1) murine embryos were co-cultured for 24 h in vitro with primary cultures of murine maternal hepatocytes (> 85% viability). Murine embryos were exposed to cyclophosphamide concentrations (0, 7.5, 15, 25 micrograms/ml), similar to those used in rat embryo culture studies. Murine embryos co-cultured with murine maternal hepatocytes developed normally, as did embryos exposed to cyclophosphamide in the absence of hepatocytes. Maternal hepatocytes were necessary for the expression of cyclophosphamide embryotoxicity, which was concentration-dependent, as demonstrated by increasing severity of reductions in crown rump length, yolk sac diameter and somite number. These results show that the co-culture of murine maternal hepatocytes and embryos is feasible, and suggest that maternal bioactivation is required for murine cyclophosphamide embryopathy. PMID- 7482568 TI - Cadmium and mercury toxicity in a human fetal hepatic cell line (WRL-68 cells). AB - The toxic effects of cadmium (Cd) and mercury (Hg), as chloride salts, were studied using an hepatic human fetal cell line (WRL-68 cells). From viability curves and the proliferative capacity of the cell in the presence of the metal, three different cell treatments were chosen, (1) 0.5 microM of the metal chloride for 24 h (acute low dose treatment), (2) 0.5 microM of the metal chloride for 7 days (chronic treatment), and (3) 5 microM of the metal chloride for 24 h (acute high dose treatment). WRL-68 cells grown in the presence of Cd exhibited the same proliferative curve as control cells, whereas in the case of Hg, the cells increased their proliferative capacity. Both metals produced ultrastructural alterations in different degrees, mainly observed as mitochondrial and RER structural changes, depending of the treatment and concentration of the metal used. Cytotoxicity was assessed by measuring the release of lactate dehydrogenase from the cells. Acutely high dose-treated cells showed the highest value for this parameter, and Cd-treated cells presented higher lactate dehydrogenase release than the Hg-treated ones. Cell damage was also measured by alanine aminotransferase (ALAT) and aspartate aminotransferase (ASAT) activities. Acute high dose Cd treatment caused the highest value of enzymatic release. Lipid peroxidation was significantly different with respect to control cells in chronic and acute high dose treatments with both metals. Metallothionein (MT) induction in response to Hg treatment was not detected. However, a dramatic induction of this protein occurred in Cd-treated cells. WRL-68 cells differentially respond to Cd and Hg making this hepatic fetal human cell line a useful tool in investigating the mechanism of toxicity of these heavy metals. PMID- 7482569 TI - Role of interferon-gamma in contact hypersensitivity assessed in interferon-gamma receptor-deficient mice. AB - The role of interferon-gamma (IFN gamma) in contact hypersensitivity induced by the haptens, oxazolone and 2,4,6-trinitrochlorobenzene (TNCB), was investigated in mice with a targeted disruption of the IFN gamma receptor (IFN gamma-R-/-). The 24-h ear-swelling response to oxazolone or TNCB in sensitized animals was not significantly reduced by the disruption of IFN gamma signalling. Dermal mononuclear infiltrates (MN) and epidermal microabscesses, however, were clearly diminished in the mutant mice. The hapten-induced upregulation of intercellular adhesion molecule-1 (ICAM-1) and major histocompatibility complex (MHC) class I in IFN gamma-R-/- mice was smaller when compared to wild-type mice. It is concluded that oxazolone- and TNCB-induced contact hypersensitivity is partially dependent on a functional IFN gamma system. While the cutaneous oedema is IFN gamma-independent, the mononuclear cell infiltration and epidermal microabscess formation are at least partly IFN gamma-dependent. Therefore, reduced cellular infiltrates are likely due to a reduced upregulation of ICAM-1 and class I antigen expression in IFN gamma-R-/- mice. PMID- 7482570 TI - Trichloroethylene--a review of the literature from a health effects perspective. AB - This report reviews the literature on the impact of exposure to trichloroethylene (TCE) on human health. Special emphasis is given to the health effects reported in excess of national norms by participants in the TCE Subregistry of the Volatile Organic Compounds Registry of the National Exposure Registries--persons with documented exposure to TCE through drinking and use of contaminated water. The health effects reported in excess by some or all of the sex and age groups studied were speech and hearing impairments, effects of stroke, liver problems, anemia and other blood disorders, diabetes, kidney disease, urinary tract disorders, and skin rashes. PMID- 7482571 TI - Reproductive toxicity screen of 1,3,5-trinitrobenzene administered in the diet of Sprague-Dawley rats. AB - Several Army installations targeted for restoration have measurable quantities of 1,3,5-trinitrobenzene (TNB) in the soil and groundwater. As part of the process of developing environmental and health effects criteria for restoration, a modified Screening Information Data Set (SIDS) reproductive study was performed. Male and female Sprague-Dawley rats received a diet containing approximately 30, 150, or 300 mg TNB/kg diet. Mating occurred following 14 days of treatment. All dams, one-half the males, and representative pups were maintained for a total of 90 days of treatment. No mortality occurred during the study; however, a decrease in mean body weights was noted in both sexes of high-dose rats. A dose-related effect was noted in measurements of sperm function/activity. Sperm depletion and degeneration of the seminiferous tubules were noted histopathologically. Methemoglobinemia and splenic hemosiderosis were common findings in the high- and mid-dose levels of both sexes at necropsy. No adverse effects were noted in mating or fertility indices. No significant treatment-related differences were found in length of gestation, sex ratio, gestation index, or mean number of pups per litter. PMID- 7482572 TI - Changes in lung permeability correlate with lung histology in a chronic exposure model. AB - In a simulated military flight-line exposure protocol, the effects of JP-8 jet fuel exposure on lung epithelial permeability were evaluated in male Fischer 344 rats (F344). Exposures were nose-only and for one hour daily. Groups were exposed for 7, 28, and 56 days. A protocol for administering a low dose (500mg/m3/hr) and a high dose (813-1094mg/m3/hr) of JP-8 jet fuel was used. Longitudinal sham exposure groups (no jet fuel) for 7, 28, and 56 days were included in the protocol. Lung epithelial permeability was measured by clearance of technetium labeled diethylenetriamine pentaacetate (99mTcDTPA, molecular weight = 492 daltons, physical half-life = 6.02 hours). The percent clearance of 99mTcDTPA per minute was calculated. Alveolar epithelial clearance for JP-8-exposed rats was dependent on both exposure concentration and duration. It was noted that at low dose exposure concentrations alveolar epithelial clearance of 99mTcDTPA returned to low levels (LD56 = 1.09% per min; LC56 = 0.98% per min), suggesting recovery as evidenced by microscopic exam. The corresponding 56-day high-dose group (n = 10) had a significantly higher (p < 0.05) value of 2.25% per minute. The 28-day low-dose (n = 15) and high-dose (n = 20) groups had clearance values that were significantly increased from their longitudinal control group (n = 17). The alveolar epithelial permeability values were 2.51, 1.95, and 1.20, respectively. The seven-day longitudinal control, low-dose, and high-dose groups had alveolar permeability values of 1.57, 2.16, and 2.07, respectively. The lung histology correlated with the clearance values. Electron micrographs showed that all groups had interstitial edema resulting from endothelial damage. There was apparent thickening of the alveolar septa, and alveolar macrophages were activated in all groups. Lung permeability data, as determined by 99mTcDTPA alveolar clearance, indicated that lung injuries peaked at 28 days of jet fuel exposure, and this finding corresponded with the histology data. There was a discrepancy in the seven-day group between the number of cells and the 99mTcDTPA clearance values. The HD7 group had a total cell count significantly higher than all other groups, but the 99mTcDTPA clearance values in that group were not significantly different from that of any other group. PMID- 7482573 TI - Evaluation of granulation peripheral blood basophils in people exposed to industrial contaminants. AB - The effect of industrial contaminants on the cytomorphometry of basophils was studied. A statistically significant difference in the densification of cytoplasmic granules of basophils was observed in people exposed to pollutants compared to people from less contaminated areas. No differences in cell area, nucleus area, or cytoplasm size were found among the tested groups. PMID- 7482574 TI - Short-term inhalation toxicity of methanol, gasoline, and methanol/gasoline in the rat. AB - Four- to five-week-old male and female Sprague Dawley rats were exposed to vapors of methanol (2500 ppm), gasoline (3200 ppm), and methanol/gasoline (2500/3200 ppm, 570/3200 ppm) six hours per day, five days per week for four weeks. Control animals were exposed to filtered room air only. Depression in body weight gain and reduced food consumption were observed in male rats, and increased relative liver weight was detected in rats of both sexes exposed to gasoline or methanol/gasoline mixtures. Rats of both sexes exposed to methanol/gasoline mixtures had increased relative kidney weight and females exposed to gasoline and methanol/gasoline mixtures had increased kidney weight. Decreased serum glucose and cholesterol were detected in male rats exposed to gasoline and methanol/gasoline mixtures. Decreased hemoglobin was observed in females inhaling vapors of gasoline and methanol/gasoline at 570/3200 ppm. Urine from rats inhaling gasoline or methanol/gasoline mixtures had up to a fourfold increase in hippuric acid, a biomarker of exposure to the toluene constituent of gasoline, and up to a sixfold elevation in ascorbic acid, a noninvasive biomarker of hepatic response. Hepatic mixed-function oxidase (aniline hydroxylase, aminopyrine N-demethylase and ethoxyresorufin O-deethylase) activities and UDP glucuronosyltransferase activity were elevated in rats exposed to gasoline and methanol/gasoline mixtures. Histopathological changes were confined to very mild changes in the nasal passages and in the uterus, where decreased incidence or absence of mucosal and myometrial eosinophilia was observed in females inhaling gasoline and methanol/gasoline at 570/3200 ppm. It was concluded that gasoline was largely responsible for the adverse effects, the most significant of which included depression in weight gain in the males, increased liver weight and hepatic microsomal enzyme activities in both sexes, and suppression of uterine eosinophilia. No apparent interactive effects between methanol and gasoline were observed. PMID- 7482575 TI - Comparative efficacy and toxicity of desferrioxamine, deferiprone and other iron and aluminium chelating drugs. AB - The efficacy and toxicity aspects of the iron and aluminium chelating drugs desferrioxamine and deferiprone (L1, 1,2-dimethyl-3-hydroxypyrid-4-one), have been compared. Major emphasis was given in the use of these two and also of other chelators in conditions of iron overload, imbalance and toxicity, as well as the incidence and possible causes of toxic side effects in both animals and humans. The chemical basis of chelation and the interaction of these chelators with the iron pools are discussed within the context of clinical application in iron overload and other conditions such as renal dialysis, rheumatoid arthritis, cancer, heart disease, malaria, etc. The design and development of new orally active alpha-ketohydroxypyridine and other chelators are considered and compared with 14 other chelators which have been previously tested in man for the removal of iron, most of which, however, were later abandoned because of low efficacy or major toxicity. The design of new therapeutic protocols based on the pharmacological, toxicological and metabolic transformation properties of the chelating drugs is also being considered, within the context of maximising their efficacy and minimising their toxicity. Overall, oral deferiprone appears to be as effective as s.c. desferrioxamine in the removal of iron and aluminium in man and to have a similar but different toxicity profile from desferrioxamine in both animals and man. The low cost and oral activity of deferiprone will make it the drug of choice for the vast majority of patients, who are not currently being chelated either because they cannot afford the high cost of desferrioxamine therapy or are not complying or have toxic side effects with its s.c. administration. PMID- 7482576 TI - Pulmonary macrophage function in systemic gadolinium chloride-pretreated rats. AB - The potential use of systemic gadolinium chloride pretreatment of rats as a tool to inhibit pulmonary macrophage function was studied by measurement of its effect on phagocytosis and nitroblue tetrazolium reduction by alveolar and interstitial macrophages. Gadolinium chloride solution was administered through the tail vein 48 h before the animals were killed. At 10 mg/kg, a dose which is generally used to inhibit Kupffer cell function, no effect on pulmonary macrophages was observed. Neither did a higher dose, 20 mg/kg, affect pulmonary macrophage function. It is concluded that gadolinium chloride pretreatment does not affect alveolar and interstitial macrophages upon systemic administration at doses and time after administration that generally depress Kupffer cell functioning. PMID- 7482577 TI - Differential inhibition of rat brain acetylcholinesterase molecular forms by 7 methoxytacrine in vitro. AB - The effects of 7-methoxytacrine (7-MEOTA), a less toxic derivative of tetrahydroaminoacridine, on the activity of acetylcholinesterase (AChE) molecular forms were investigated in vitro. AChE molecular forms were separated by sucrose gradient sedimentation from homogenates of the frontal cerebral cortex prepared with buffer containing Triton X-100 (soluble + membrane-bound enzyme). Two molecular forms, namely 10S and 4S corresponding to globular tetrameric (G4) and monomeric (G1) forms, respectively, were detected; their molecular weights were 220,000 and 54,000 Da. A significantly higher sensitivity to 7-MEOTA of G4 than of G1 forms was observed. The Ki values were 0.21 +/- 0.07 microM for the former and 0.70 +/- 0.15 microM for the latter. The differential inhibition of AChE molecular forms by 7-MEOTA is discussed in relation to its possible clinical application for treatment of disorders such as Alzheimer's disease, in which a reduction of brain cholinergic neurotransmission is believed to play a role. PMID- 7482578 TI - Selective promotion by phenylmethanesulfonyl fluoride of peripheral and spinal cord neuropathies initiated by diisopropyl phosphorofluoridate in the hen. AB - This paper reports studies in hens showing that diisopropyl phosphorofluoridate (DFP) neuropathy is promoted by PMSF when initiated either in central (spinal cord) or peripheral nervous system. Moreover, the critical site for promotion is in peripheral nerve axons rather than in their cell bodies. Selective promotion in peripheral nerves was achieved by giving PMSF into sciatic artery monolaterally (7 mg/kg) to birds where neuropathy was initiated by DFP, either systematically (0.3 mg/kg s.c.) or intra-arterially (0.04 mg/kg in the same artery). Birds developed monolateral neuropathy in the leg where PMSF was delivered. Promotion of spinal cord neuropathy was achieved by giving PMSF (120 mg/kg s.c.) to birds where neuropathy was initiated selectively in spinal cord. This was obtained by protecting peripheral axons with intra-arterial bilateral injections of PMSF (0.55 x 2 mg/kg) followed by DFP (0.3, 0.4 or 0.7 mg/kg s.c.). The resulting syndrome was characterized by spastic ataxia. PMID- 7482579 TI - Nephrotoxic potential of N-(3,5-dichlorophenyl)glutarimide and N-(3,5 dichlorophenyl)glutaramic acid in Fischer 344 rats. AB - The agricultural fungicide N-(3,5-dichlorophenyl)succinimide (NDPS) produces kidney damage in rats. Although many NDPS analogues have been screened as possible nephrotoxicants, the one-carbon homologue, N-(3,5 dichlorophenyl)glutarimide (NDPG), has not been evaluated. This study examined the nephrotoxic potential of NDPG and a putative metabolite, N-(3,5 dichlorophenyl)glutaramic acid (NDPGA). Male Fischer 344 rats (N = 3-4 per group) were administered a single i.p. injection in corn oil of NDPG or NDPGA (0.4 or 1.0 mmol/kg), NDPS (0.4 mmol/kg), or corn oil alone. Renal function was monitored for 48 h. In contrast to NDPS, NDPG and NDPGA did not significantly alter renal function or kidney morphology when compared to corn oil-treated controls. These experiments show that replacement of the succinimide ring in NDPS with a glutarimide ring abolishes toxicity. PMID- 7482580 TI - The effects of 2,2',4,4',5,5'-hexachlorobiphenyl cotreatment on the disposition of 2,3,7,8-tetrachlorodibenzo-p-dioxin in mice. AB - Two groups of C57BL/6J mice received a single oral dose of 1 nmol/kg 2,3,7,8 [3H]tetrachlorodibenzo-p-dioxin (TCDD) alone or in combination with 300 mumol/kg 2,2',4,4',5,5'-hexachlorobiphenyl (HxCB). The disposition of TCDD in liver, fat, skin, spleen, lung and blood was studied at days 3, 7, 13 and 34 after dosage. HxCB cotreatment increased hepatic TCDD levels and, consequently, significant increases of the liver/fat distribution ratio were observed. HxCB cotreatment did not significantly affect TCDD levels in fat or other tissues. The elimination rates of TCDD were not influenced by HxCB cotreatment in any of the tissues. It is concluded that HxCB cotreatment alters the body distribution of TCDD in mice but does not influence the elimination rate of TCDD. PMID- 7482581 TI - Effects of 3'-azido-3'-deoxythymidine on erythroid inducible gene expression in human K-562 leukemia cells. AB - We previously demonstrated that 3'-azido-3'-deoxythymidine (AZT) down regulates hemoglobin (Hb) synthesis and globin gene expression. That inhibition may therefore result either from a direct effect on globin gene transcription or an indirect effect through inhibition of K-562 cell induction, thereby leading to inhibition of other inducible genes of heme biosynthesis. The present results demonstrate that inhibition of globin gene expression by AZT is a direct gene effect rather than a general inhibition of K-562 cell induction as demonstrated by the absence of AZT effects on expression of three other erythroid inducible genes [erythroid-specific aminolevulate synthase (ALAS-E), aminolevulinic acid dehydrogenase (ALAD), and erythroid-specific porphobilinogen deaminase (PBGD-E)]. PMID- 7482582 TI - Induction of protein droplet (alpha 2 mu-globulin) nephropathy in male rats after short-term dosage with 1,8-cineole and l-limonene. AB - As part of a series of short-term studies on peppermint oil constituents for their possible induction of the encephalopathy found with peppermint oil, 1,8 cineole and l-limonene were studied. Groups of 10 male Wistar rats were given 0, 500, or 1000 mg 1,8-cineole/kg body wt./day or 0, 800, or 1600 mg l-limonene/kg body wt./day for 28 days. 1,8-Cineole and l-limonene both induced accumulation of protein droplets containing alpha 2 mu-globulin in proximal tubular epithelial cells in male rats. These results suggest that both 1,8-cineole and l-limonene possibly belong to the group of chemicals characterized by their induction of excessive alpha 2 mu-globulin accumulation. Neither of the 2 substances induced encephalopathy. PMID- 7482583 TI - Inhibitory effect of steviol, a metabolite of stevioside, on glucose absorption in everted hamster intestine in vitro. AB - The effects of stevioside and steviol (a product of enzymatic hydrolysis of stevioside) on intestinal glucose absorption were examined in hamster jejunum. By using the everted sac technique, we found that stevioside (1 and 5 mM) had no inhibitory effect on glucose absorption. In contrast, glucose absorption was inhibited 29% by 1 mM steviol. The inhibition of glucose absorption by steviol was related to steviol concentration and incubation time. The possible mechanism of steviol inhibitory action of glucose absorption was also investigated. Reductions in the intestinal mucosal ATP content and absorptive surface area were responsible for the inhibition of glucose absorption by steviol. The decrease in the intestinal mucosal ATP content was accompanied by a decrease in the activities of mitochondrial NADH cytochrome c reductase and cytochrome oxidase. Moreover, no inhibitory effects of steviol on the activity of intestinal Na+,K(+) ATPase and glucose uptake in the intestinal brush-border membrane vesicles were seen. These results suggest that inhibition of intestinal glucose absorption by steviol in hamsters is due to the reduction in mucosal ATP content and an alteration of the morphology of the intestinal absorptive cells. PMID- 7482585 TI - The percutaneous absorption of hydroquinone (HQ) through rat and human skin in vitro. AB - Because of the potential for human contact with photographic developer solutions containing hydroquinone (HQ), the rates of percutaneous absorption of HQ through human stratum corneum and full-thickness rat skin have been measured in vitro using 5% aqueous solutions of HQ as the donor solutions. The studies were performed using infinite doses of aqueous solutions containing 14C-labeled HQ in Franz-type diffusion cells. The measured absorption rate (mean +/- S.D.) of HQ through human stratum corneum was 0.52 +/- 0.13 micrograms/cm2/h, while that for full-thickness rat skin was 1.1 +/- 0.65 micrograms/cm2/h. The ratio (rat/human) of the permeability constants (Kp) was 2.4. Using the definitions suggested by Marzulli et al. (1969) Toxicol. Appl. Pharmacol. Suppl. 3, 76-83, HQ would be classified as 'slow' with respect to its absorption through human stratum corneum. PMID- 7482584 TI - Direct effect of cadmium on citrate uptake by isolated rat renal brush border membrane vesicles. AB - High incidence of multiple kidney stone formation has been observed among workers exposed to cadmium (Cd). Citrate is known to be a protective factor against renal stone formation. To study the direct effect of cadmium on citrate uptake by the renal brush border membrane, we exposed isolated rat renal brush border membrane vesicles (BBMV) to cadmium and determined their citrate uptake characteristics. BBMV were prepared by the divalent cation precipitation method. Citrate uptake was measured by the Millipore rapid membrane filtration technique. Preincubation of BBMV with 2 and 10 mM CdCl2 for 1 min significantly inhibited citrate uptake compared with that of BBMV without Cd. Analysis of the time course of citrate uptake during 30-min preincubation of BBMV with 0.5 mM Cd also revealed significant reduction of the uptake compared with that of the control BBMV without preincubation. These findings indicate that preincubation of BBMV with cadmium results in time-dependent and concentration-dependent inhibition of citrate uptake. PMID- 7482586 TI - Beware of a new carcinogenic risk: promotion of promoters. AB - Restricting the evaluation of carcinogenic risk to genotoxicity or mutagenicity tests, as recently recommended by some researchers, is a hazardous operation, since the carcinogenic process involves a complex sequence of events in which no simple, necessary and sufficient condition is known for either the occurrence or non-occurrence of a cancer. None of the available tests exhibit a generalizable meaning, while none of the carcinogenic process phases should be neglected in prediction studies. In particular, ignoring promoter action and progression or proliferation, as well as targets specificity and other points, should lead to an underestimation of the actual risk. PMID- 7482587 TI - Correlation between cumulative labeling indices measured in the terminal bronchioles and in the centriacinar region in the lungs of rats exposed to oxidant air pollutants. AB - Male Sprague-Dawley rats were exposed on 7 consecutive days for 8 h a night to 0.6 ppm ozone, 10.8 ppm NO2 or a mixture of the two gases. Cumulative labeling indices were measured in the epithelium of the terminal bronchioles and the adjacent centriacinar region. An excellent correlation in labeling indices between the two sites was found. It is concluded that the labeling index in the terminal bronchioles represents a sensitive measurement for ozone-induced lung injury. PMID- 7482589 TI - Effects of pyridoxal 5'-phosphate on plasma alanine aminotransferase determinations in toxicological studies. AB - Alanine aminotransferase activities (ALT) were measured in rat plasma samples with three different reagent formulations. The inclusion of pyridoxal 5' phosphate in the reagent produced higher ALT values, and altered the statistical relationships between control and treatment groups in three studies where plasma ALT values were reduced by xenobiotics. PMID- 7482588 TI - Effect of atropine on cyanide-induced acute lethality in mice. AB - The effects of atropine on acute lethality induced by cyanide were investigated in mice. The LD50 value of cyanide (s.c. injection) was 8.4 (7.6-9.3) mg/kg. However, the LD50 value of cyanide (s.c.) was significantly increased by 1.5-fold when atropine (32 mg/kg) was injected s.c. in mice. Furthermore, the combined administration of atropine (32 mg/kg), Ca2+ (500 mg/kg) and sodium thiosulfate (1 g/kg) tremendously increased the LD50 value by 5.6-fold in mice although sodium thiosulfate or Ca2+ alone increased the LD50 2.5- or 1.5-fold. On the other hand, although the LD50 value of cyanide (intracerebroventricular injection (i.v.t.)) was 52.0 (47.4-57.0) micrograms/brain, the LD50 value of cyanide (i.v.t.) was significantly increased by 1.3- or 1.61-fold in mice 10 min after s.c. injection of atropine (32 mg/kg) or Ca2+ (500 mg/kg). Furthermore, the combined administration of atropine and Ca2+ increased the LD50 value of cyanide by 2.1 fold. These results suggest that atropine inhibits cyanide-induced acute lethality and promotes the antagonistic effect of thiosulfate and Ca2+ in mice. PMID- 7482590 TI - Three weeks' and six months' exposure to aromatic white spirit affect synaptosomal neurochemistry in rats. AB - The effects of 3 weeks' or 6 months' inhalation exposure of rats to aromatic white spirit 6 h/day, 5 days/week at 0, 400, or 800 ppm were studied. Synaptosomal neurochemistry was investigated as index of the in situ conditions in the presynaptic nerve terminal. In both studies, the relative and absolute yield of synaptosomal protein were significantly reduced in the two exposed groups. Both studies demonstrated increased synaptosomal noradrenaline (NA), dopamine (DA), and 5-hydroxytryptamine (5-HT) concentrations, high- affinity 5-HT uptake rate and uptake capacity. It is hypothesized that a reduced density and total number of synapses in situ may be functionally compensated by increased NA, DA, and 5-HT neurotransmitter release, or by increased activity of corresponding neurons. The increased synaptosomal 5-HT uptake rates and uptake capacities may explain the previously demonstrated increased global and regional neurotransmitter concentrations and the present finding of increased synaptosomal 5-HT concentrations. These changes are interpreted as an indication of toxic effect on the CNS function and are considered supportive of recent findings of electrophysiological changes and affected motor activity following 6 months' exposure to dearomatized white spirit followed by an exposure-free period. PMID- 7482591 TI - Significance of elevated urinary human intestinal alkaline phosphatase in Japanese people exposed to environmental cadmium. AB - Urinary human intestinal alkaline phosphatase (IAP), beta 2-microglobulin (beta 2 MG) and N-acetyl-beta-D-glucosaminidase (NAG) were analyzed in 40 Japanese environmental-cadmium (Cd)-exposed and 40 non-exposed subjects to evaluate early biological markers for Cd-induced renal damage. All urinary indicators were significantly higher in the Cd-exposed subjects than non-exposed subjects. A fourth-order function was fitted for the relationship between beta 2-MG and IAP or NAG. The beta 2-MG concentration corresponding to the inflexion point for IAP was smaller than that for NAG. This result may support the contention that the cells containing IAP are damaged earlier than those containing NAG, and that IAP is a useful marker for detecting renal tubular dysfunction in people moderately exposed to Cd. However, in the stage of severe renal damage, the combination of IAP and beta 2-MG is considered to be more useful. PMID- 7482592 TI - Activity of some respiratory enzymes and cytochrome contents in rat hepatic mitochondria following aflatoxin B1 administration. AB - Effects of aflatoxin B1 (AFB1) administration (7 mg/kg body weight, i.p.) on rat hepatic mitochondrial respiratory components have been examined. Succinoxidase and cytochrome oxidase activities were decreased in liver mitochondria isolated from rats 12-24 h after AFB1 treatment. Both enzyme activities returned to normal levels after 48 h. Glutamate dehydrogenase and beta-hydroxybutyrate dehydrogenase activities did not show any alterations up to 24 h and thereafter increased at 48 72 h. Succinate dehydrogenase activity was impaired by 41% at 12 h and thereafter was found to be normal. The intramitochondrial cytochrome b content declined at 24-72 h, whereas cytochrome aa3 content was decreased maximally at 72 h after AFB1 administration. These observations on mitochondrial enzyme activities and cytochrome contents correlate well with our earlier observations made on hepatic mitochondrial respiratory rates after AFB1 treatment. The impairment of respiratory functions possibly results from membrane damage and selective modification of gene expression in mitochondria imparted by AFB1. PMID- 7482594 TI - A quantitative structure-activity relationship for the eye irritation potential of neutral organic chemicals. AB - Quantitative structure-activity relationships (QSARs) have been derived relating eye irritation data of a set of neutral organic chemicals to log(octanol-water partition coefficient), the minor principal inertial axes (Ry and Rz) and dipole moment. Datasets were analysed using principal components analysis; plots of the first 2 principal components of the above parameters showed that the analysis was able to discriminate well between the irritant and non-irritant chemicals in the dataset. The derived QSAR could be useful for the prediction of the eye irritation potential of new or untested chemicals within this category. PMID- 7482593 TI - Safety evaluation of benzophenone-3 after dermal administration in rats. AB - Benzophenone-3 (BZ-3) is a category 1 (over-the-counter) product approved by the US Food and Drug Administration (FDA) for use as a sunscreen agent in medicine, cosmetics, industry, and agriculture. This is due to its ability to absorb and dissipate ultraviolet light in a harmless manner, thus protecting human skin and products from UV irradiation. This study investigated the safety of BZ-3 after repeated administration. BZ-3 in ointment base was applied at a dose of 100 mg/kg body wt. twice daily, for 4 weeks to the skin of male Sprague-Dawley rats. Body weight, organ to body weight ratios, hematological, and clinical chemistry parameters were not effected. Pathological examination revealed no significant changes between control and treated animals. No gross external abnormalities were observed. Both in vivo and in vitro blood glutathione (GSH) levels were effected by BZ-3 treatment. However, after 60 min of incubation, a reversal of this effect was observed in the treatment group as blood GSH levels approached normal levels. Furthermore, investigation of GSH-reductase and peroxidase with time indicated an increase in GSH-reductase activity at 60 and 90 min with no effect on GSH peroxidase. Pre-treatment with phenobarbital modulated the metabolic disposition of BZ-3. There was an increase in the formation of the hydroxy metabolites but not the O-dealkylated form. This study suggests that BZ-3 is not toxic to rats when applied dermally at a dose of 100 mg/kg body wt. for 4 weeks. PMID- 7482595 TI - Inhibition of P-450 by aucubin: is the biological activity of aucubin due to its glutaraldehyde-like aglycone? AB - The inhibition of ethoxy coumarin O-deethylase (ECOD) activity by aucubin and its aglycone was examined in a microsomal system and in freshly isolated hepatocytes. Aucubin was found to be inactive but the aglycone was found to be a potent time dependent inhibitor of ECOD activity in both systems. The close structural similarity between the aglycone of aucubin and glutaraldehyde suggests a similar mechanism of enzyme inhibition through protein cross-linking by Schiff reactions. The similarity between the 2 compounds was demonstrated through their closely similar binding spectra to bovine serum albumin. The biological activities reported for the aglycone are suggested to be due to this similarity to glutaraldehyde. PMID- 7482596 TI - Fosinopril treatment of pregnant rats: developmental toxicity, fetal angiotensin converting enzyme inhibition, and fetal angiotensin II receptor regulation. AB - Pregnant women are advised against using angiotensin-converting enzyme (ACE) inhibitors due to reports of adverse effects on human fetuses. This study examined ACE binding and angiotensin II (Ang II) receptor binding in fetuses of rats treated with the ACE inhibitor fosinopril (16 mg/kg/day fosinopril, p.o. in 4 divided doses, from gestational day (gd) 13 to gd 18). Binding of the potent radiolabeled ACE inhibitor 125I-351A to ACE in the lung and aorta of gd 19 fetuses of fosinopril-treated dams was reduced by 56 and 44%, respectively, compared to fetuses from vehicle-treated dams, indicating that fosinopril or its active metabolite, fosinoprilat, crosses the placental barrier and inhibits fetal ACE. Fetal Ang II receptor binding of 125I-Sar1,Ile8 Ang II was not altered in most of the tissues examined, although reductions in binding in the adrenal of fetuses of fosinopril-treated dams approached statistical significance. PMID- 7482597 TI - Administration of subtoxic doses of t-butyl alcohol and trichloroacetic acid to male Wistar rats to study the interactive toxicity. AB - Tertiary butyl alcohol and trichloroacetic acid (TCA) are known to be contaminants in drinking water. In order to evaluate the interactive toxicity of t-butyl alcohol (TBA) with TCA, young male Wistar rats were dosed through water at a dose level of TBA (0.5% v/v), 25 ppm TCA and a combined dose of TBA+TCA (0.5% v/v TBA, 25 ppm TCA) for a period of 10 weeks ad libitum and were maintained on normal diet. The control animals received plain water and normal diet. There was remarkable loss of body weight and significantly decreased liver triglycerides in the treatment groups in the order of TBA+TCA, TCA, TBA and increased liver weights were observed. Serum succinate dehydrogenase (SDH) levels were significantly increased in TCA- and TBA+TCA-treated groups. There was no significant change in serum alanine (GPT), aspartate (GOT) aminotransferase, serum alkaline (ALP) and acid (ACP) phosphatase levels as well as liver glutathione (GSH) and liver and serum cholesterol levels in the treated groups. But serum triglycerides, liver glycogen, serum glucose (only in TBA- and TCA treated animals) were significantly high in the treated groups. Lipid peroxidation measured by diene conjugation was significant in TBA+TCA-treated group and kidney GSH levels were significantly low in the treated groups. These results show that interaction of TBA+TCA does bring about alteration in biochemical parameters which may play a pivotal role in toxic responses on long term exposure. PMID- 7482598 TI - [Morphological and experimental studies of the autonomic ganglia of the head]. PMID- 7482599 TI - [The free-radical involvement of the salivary gland in stress]. AB - It has been demonstrated in rat experiments that acute emotional painful stress enhanced free-radical lipid oxidation in salivary gland tissues and reduced the activities of antioxidant enzymes in it. Similar processes take place in the blood. Antiaggregation activity increases in the tissues of salivary gland, this being an essential adaptation factor in stress, preventing microcirculatory disorders in the tissues. PMID- 7482600 TI - [The effect of mineral and organic substances on the desorption of sugar from a soft dental deposit]. AB - Effects of the principle mineral and mineralizing components of the saliva, alimentary organic acids-products of carbohydrate metabolism, and of some other substances of various pH on sugar desorption from soft dental deposit were compared in vitro. The desorbing efficacy was assessed from the concentration of sucrose, derived from a sample of lyophilized dental deposit, in a suspension of the tested reagent. The major mineral components of the saliva, calcium and phosphate, were found to promote extraction of sucrose from the deposit in neutral and weakly alkaline media. Acid medium also favors sucrose desorption. Due to the physiological nature and efficacy of sucrose desorption, solutions of alimentary organic acids may be recommended for prevention of cariogenic situations caused by intake of easily fermented carbohydrates. PMID- 7482601 TI - [The nature of the primary interaction of saccharose from aqueous solutions with the oral cavity tissues]. AB - The course of saccharose adsorption on surface tissues of the oral cavity directly after carbohydrate loading was studied in vivo. The amount of saccharose adsorbed on tissue surface was assessed from its desorption from an isolated site of the solid phase of the oral cavity and was correlated to a unit of tissue surface. Cariogenic potential in the oral cavity was shown to be maintained to a great measure at the expense of soft tissues and, specifically, of the glossal mucosa which is characterized by a high adsorption capacity for saccharose and represents a sort of a saccharose "depot" in the oral cavity. Carbohydrate adsorption appears to be a physical process. Water gargle after sugar intake fails to solve the problem of prevention of a cariogenic situation. Weakly acid gargling appears to be more promising desorbents of saccharose from oral tissues. PMID- 7482603 TI - [A modified toothbrush for professional oral hygiene]. PMID- 7482602 TI - [The enzymes of glutamate and organic phosphate metabolism in the saliva in fluorosis (clinical and experimental research)]. AB - Activities of glutamic-oxalacetic transaminase, glutamate dehydrogenase, gamma glutamyl transferase were reduced in unstimulated pooled saliva of 15-year-old adolescents with endemic fluorosis, vs. those without fluorosis. This point to reduced production of glutamate which is indispensable for bacterial growth. The activity of acid phosphatase in pooled stimulated with 1% pilocarpin saliva of rats fed a water ration with 5 and 20 mg/liter fluorine in comparison with control rats fed water with fluorine concentration of 0.21 mg/liter. Noteworthy that alkaline phosphatase activity was virtually unchanged at fluorine concentrations 5 and 20 mg/liter in the water. The significance of the results as far as it regards the pathogenesis of caries and fluorosis is discussed. PMID- 7482604 TI - [The periodontal status of patients with hypoestrogenemia in relation to the severity of systemic osteoporosis]. AB - Changes in the periodontium progress in the patients with hypoestrogenemia, this progress being the slowest in patients with amenorrhea, intermediate in those subjected to oophorectomy, and the most rapid in those in the postmenopausal period. A conclusion was made that reduction of mineralisation of the bones enhances pathologic changes in the periodontium. PMID- 7482605 TI - [A clinical x-ray evaluation of the efficacy of hydroxyapatite with collagen in treating periodontitis and radicular cysts]. AB - One-year follow-up of 20 patients with radicular cysts and 16 ones with medium severity chronic periodontitis demonstrated the efficacy of hydroxylapatite with collagen. Active formation of osseous tissue at the site of cysts, in the periodontal osseous pockets, and at the apexes of interdental septa was observed 6 to 7 months after the treatment. PMID- 7482606 TI - [Immunocorrective therapy in the treatment of chronic herpetic stomatitis by using magnetic autohemotherapy]. AB - Twenty-seven patients with chronic herpetic stomatitis were treated using routine protocols including traditional autohemotherapy (controls, n = 12) and magnetic autohemotherapy (main group, n = 15) developed by the authors. Local therapy was identical in both groups and corresponded to the disease stage. Treatment with magnetic autohemotherapy proved to be highly effective: it prolonged the remission and reduced the incidence and duration of exacerbations. Cytological findings indicate that such an exposure activated the processes of regeneration of the buccal and labial mucosal epithelium. A trend to normalization of some parameters of cellular and humoral immunity was observed. PMID- 7482607 TI - [The role of plasmosorption in correcting the microcirculatory disorders in patients with suppurative-inflammatory processes of the maxillofacial area and diabetes mellitus]. AB - A positive clinical effect of plasma sorption added to multiple-modality treatment of patients with pyoinflammatory maxillofacial diseases and diabetes mellitus was revealed. Plasma sorption is conducive to normalization of lipid metabolism, improves blood rheology, activates microcirculation, this promoting sooner healing of postoperative wounds and reducing hospitalization periods. PMID- 7482608 TI - [The involvement of the temporomandibular joints in systemic lupus erythematosus]. AB - Seventy-three patients with systemic lupus erythematosus were examined, in 55 (75%) of whom involvement of the temporomandibular joint was detected, this involvement occurring at the debut of the disease, before involvement of other joints, in 44 subjects. The disease presented as ephemeral migrating asymmetrical arthralgia, more expressed by the evening hours, and, rarely, as temporomandibular arthritis and painful (functional) contractures. A frequent dissociation between the clinical and x-ray signs of the disease, no residual phenomena were observed. X-Ray examination can effectively rule out other diseases of the temporomandibular joint, first of all, destructive genesis. PMID- 7482609 TI - [The mechanisms of the cytoprotective action of prostacyclin in acute postoperative parotitis]. AB - Effects of prostaglandin I2 of the content of lipid peroxidation products and the activity of glutathione antioxidative system were studied in 60 dogs in whom acute nonepidemic parotitis was induced. Prostaglandin I2 was found to enhance the reactions of protective inhibition of metabolism with reduction of peroxidation level and activation of the antioxidative system, this permitting the cell to preserve its structural intactness. A conclusion is made about the possibility of using prostaglandin I2 for the prevention and therapy of acute nonepidemic parotitis. PMID- 7482610 TI - [Epithelial tumors of the minor salivary glands in the lips and cheeks]. AB - Difficulties in the clinical diagnosis of tumors of minor salivary glands of the lips and cheeks are discussed on the basis of results of examination of 52 patients with such tumors and comparison of the diagnosis of this condition and of identical tumors of the large salivary glands. Discrepancies between the clinical and pathoanatomical diagnosis are analyzed. Clinical characteristics helpful in the diagnosis of the condition are presented. PMID- 7482611 TI - [The treatment of acute periodontitis using sodium diclofenac experimentally]. PMID- 7482612 TI - [A computer program for the rapid luminescent cytological diagnosis of precancer and cancer of the lip and mouth mucosa]. AB - An algorithm has been derived and a computer program based on it developed, which helps to the dentist to carry out fluorescent cytological investigations and make a tentative rapid diagnosis using a PC. The algorithm is based on a simple standard assessment of the cytological preparation. Efficacy of differential diagnosis of labial and buccal cancer and precancer is 91.9 +/- 3.2%. PMID- 7482613 TI - [The choice of the method for surgical intervention in the combined treatment of cancer of the oral mucosa in the retromolar space]. AB - Combined treatment of 30 patients with stage III-IV squamous-cell carcinoma of the retromolar mucosa, including radiotherapy in a dose of 43 Gy at the first stage of treatment, was carried out. The second stage consisted of two types of combined operations. Modified combined intervention suggested by P. G. Bitiutskii et al., the results of which were followed up for 2 years and found satisfactory, is preferable to previously employed operation, for it is conducive to a more rapid functional rehabilitation of patients. PMID- 7482614 TI - [Transtracheal jet pulmonary ventilation as a measure for preventing hypoxia during reconstructive maxillofacial operations]. AB - Anesthesia including high- or low-frequency transtracheal jet ventilation of the lungs was used in 52 patients with maxillofacial abnormalities subjected to repair surgery. The number of respiratory cycles was 20 per min in 1 group and 120 per min in the other. Intubation of the trachea was started 20 min after ventilation was started. Transtracheal jet ventilation of the lungs prevented hypoxia during intubation of the trachea in such patients. High-frequency jet ventilation of the lungs may be a method of choice in such cases. PMID- 7482615 TI - [The efficacy of using metal ceramic dentures fixed on pulpectomized teeth]. AB - A total of 432 patients aged 34 to 60 were examined, 226 of them aged 31 to 59 were admitted for treatment. Out of 1980 examined cermet crowns fixed on depulped teeth complications were detected round 412 (20.88%). Inflammations of the gingiva were found near 218 crowns, apical chronic periodontitis in 98 cases, functional over-exercise of the periodontium in 61, ceramic fracture in 18, failure of cement fixation in 17 cases. When all prophylactic measures are taken, apical periodontitis was properly treated, and dental channels filled during pulp removal, if indicated, complications develop almost three times less frequently, occurring in 7.4% of cases. PMID- 7482616 TI - [Plasma technology in the practice of orthodontics. 1]. AB - The paper describes the technique of plasma spraying used to fortify the coating of permanent dentures. This method helps improve the quality of permanent dentures and strengthen the adhesion of coatings on retention layers. Hence, dentures with very thin retention layer and vary strong coating adhesion can be made by traditional methods of denture making, thus improving the quality of prosthetic treatment. Traditional plastic may be used as coating material. PMID- 7482617 TI - [Methods for examining children with sialolithiasis of the submandibular glands]. PMID- 7482618 TI - [The comparative characteristics of the transfocal osteosynthesis of mandibular fractures with devices made of titanium nickelide and a wire suture]. PMID- 7482619 TI - [The periodontium and mouth mucosa in patients with neutropenia]. PMID- 7482620 TI - [Salivary gland heterotopia in the bone tissue of the mandible]. AB - A rare case, heterotopia of the salivary gland in a female patient with repeated recurrences of radicular cysts of the mandible, is described. Histologic examination of removed material revealed dystopic acinar tissue in the subepithelial layer of the radicular cyst. A hypothesis on the possible effect of acinar heterotopic tissue on recurrences of the cystic process is offered. PMID- 7482621 TI - [Cases of the detection by the dentist of syphilis in children]. PMID- 7482622 TI - [Experience in teaching biochemistry to the students of a dentistry department]. PMID- 7482623 TI - [The ultrasonic diagnosis of soft-tissue diseases of the maxillofacial area]. PMID- 7482624 TI - [Morphological changes in the periodontium from the use of different designs of metal ceramic dentures]. AB - Morphological changes in the periodontium during the use of cermet crowns of different design were studied in experimental animals. The status of the periodontium was found to depend on the level of crown fixation with respect to gingival edge. Histological findings had show that position of a cermet crown at the level of the gingival edge does not involve appreciable changes in the periodontium, though preparation of abutment teeth with the formation of a step be considered as preferable. A subgingival position of cermet crowns leads to development of an inflammatory reaction in the periodontium, the severity of this reaction being related to a great measure to the method of the tooth preparation. If a step is formed during preparation of a tooth, the inflammatory changes are less pronounced than when there is no step formed. In the latter case the pathological changes were seen in the periodontium. The authors come to a conclusion that cermet crowns should be fixed at the level of the gingival edge. When making cermet crowns, abutment teeth should be prepared with the formation of a pericervical step. PMID- 7482625 TI - A decreasing CD4+/CD8+ ratio after one month of treatment with stanazolol in postmenopausal women. AB - Androgens influence some immunological processes, including the differentiation of T-cells in CD4+ (helpers) or CD8+ (suppressors/cytotoxic) phenotype. In nine postmenopausal osteoporotic women the effect of stanazolol on lymphocyte counts, CD3+ and the immunoregulatory index (CD4+/CD8+) were investigated. In the placebo group, ten postmenopausal osteoporotic women of similar age were included. The means of the investigated indices after stanazolol as compared with the values before treatment were as follows: lymphocyte counts (cells/microL +/- SEM) 2974 +/- 225 versus 2313 +/- 166, CD3+ (%) 54.3 +/- 5.5 versus 70.9 +/- 1.6 (P < 0.05); CD4+/CD8+ ratio 1.8 +/- 0.02 versus 2.5 +/- 0.28 (P < 0.05). The values after placebo as compared with the values before placebo were: 2558 +/- 201 versus 2370 +/- 256, 62.9 +/- 2.1 versus 64.8 +/- 1.7 and 1.6 +/- 0.2 versus 1.6 +/- 0.1 in sequence. The treatment was controlled by the serum stanazolol levels before and after steroid administration (unmeasurable versus 20.8 +/- 3.4 nmol/L, P < 0.01). The good compliance of the therapy was confirmed by a decline of serum LH (U/L; 30.1 +/- 3.1 versus 24.7 +/- 2.8, P = 0.014), FSH (U/L; 108.9 +/- 13.1 versus 93.3 +/- 12.8, P = 0.012) and serum sex hormone binding globulin (SHBG; nmol/L; 53.3 +/- 13.3 versus 11.2 +/- 1.9, P < 0.01). The decline of SHBG indicates a good tissue sensitivity to the androgen. There were no significant differences between hormonal parameters before and after placebo treatment. In conclusion, the immunosuppressive effect of the androgen, stanazolol, was confirmed in the investigated postmenopausal osteoporotic women.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7482626 TI - Cleavage reaction of 5 beta, 6 beta steroidal epoxides carrying different groups at C-3. AB - The particular behavior of 5 beta, 6 beta steroidal epoxides carrying different groups at C-3 was studied. These epoxides may exhibit different cleavage behavior according to the nature of the solvent, the acid-base state of the medium, and the leaving abilities of the C-3 substituent. Results and alternative mechanisms are presented. PMID- 7482627 TI - Synthesis of 3 alpha, 7 alpha, 12 alpha-trihydroxy- and 3 alpha, 7 alpha dihydroxy-5 beta-cholestan-26-oic acids by the use of beta-ketosulfoxide. AB - The biosynthetic intermediates of bile acid, 3 alpha, 7 alpha, 12 alpha trihydroxy- and 3 alpha, 7 alpha-dihydroxy-5 beta-cholestan-26-oic acids, were synthesized by means of the thermal elimination of beta-ketosulfoxides. The alpha, beta-unsaturated ketones as key compounds of the synthesis, 3 alpha, 7 alpha, 12 alpha-trihydroxy- and 3 alpha, 7 alpha-dihydroxy-5 beta-cholest-25-en 24-ones, were effectively derived from the beta-ketosulfoxides prepared from methyl cholate or chenodeoxycholate by reaction with methylsulfinylcarbanion. These unsaturated ketones were converted into 3 alpha, 7 alpha, 12 alpha, 26 tetrahydroxy- and 3 alpha, 7 alpha, 26-trihydroxy-5 beta-cholestanes by reductive deoxygenation and hydroboration, of which stereoisomers were chromatographically separated into 25S- and 25R- isomers. The oxidation of each of the above isomeric alcohols after the protection of the hydroxyl groups on the steroidal ring and the following hydrolysis gave the title 26-carboxylic acids. PMID- 7482628 TI - Steroidal anti-inflammatory antedrugs: synthesis and pharmacological evaluation of 16 alpha-alkoxycarbonyl-17-deoxyprednisolone derivatives. AB - In a continuing effort to minimize the systemic adverse effects of potent anti inflammatory steroids, a series of 16 alpha-alkoxycarbonyl-17-deoxyprednisolone derivatives: methyl (8a), ethyl (8b), isopropyl (8c), and benzyl (8d) 11 beta,21 dihydroxy-3,20-dioxo-1,4-pregnadiene-16 alpha-carboxylate, was synthesized and evaluated for their topical and local anti-inflammatory activities. In the acute croton oil-induced ear edema dose-response bioassay, the topical anti inflammatory potencies of these esters relative to prednisolone, 1, were: 8a:1.0, 8b:1.3, 8c:4.0, 8a:4.7 and 1:1.0. The putative metabolite, 11 beta,21-dihydroxy 3,20-dioxo-1,4-pregnadiene-16 alpha-carboxylic acid, 7, was inactive in this test. A seven day cotton pellet granuloma bioassay was employed to study the local and systemic anti-inflammatory activities of these steroids. The local anti inflammatory potencies of these esters relative to prednisolone, 1, were 1.3, 1.5, 2.3, 2.5, and 1.0 for 8a, 8b, 8c, 8d, and 1, respectively. In this semi chronic study, only prednisolone exhibited significant untoward side effects, such as reduction in thymus weights, normal body weight gain, and normal plasma corticosterone levels. The increase in the topical and local potencies of these steroid esters was consistent with the increase in their 1-octanol/buffer partition coefficient. The ratio of local to systemic anti-inflammatory activity of 8c and 8d was four times that of prednisolone. The effects of increasing the size of the alkoxy group of these new steroids on both topical and local anti inflammatory activity and their concomitant decrease in untoward systemic effects were unequivocally demonstrated.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7482630 TI - Formation of 6 beta-OH-deoxycorticosterone from deoxycorticosterone by A6 cells. AB - We have investigated the metabolism of deoxycorticosterone in A6 cells, a continuous cell line derived from the kidney of Xenopus laevis. A6 cells at confluence were incubated with serum-free culture media containing 2.5 microM [3H] deoxycorticosterone. When radioactive compounds in incubation media were separated by reverse-phase high-pressure liquid chromatography, the formation of polar metabolites was observed. One component of polar metabolites cochromatographed with 6 beta-hydroxydeoxycorticosterone. In order to identify this component more rigorously, large scale cultures were performed and this compound was separated and purified by reverse-phase high-pressure liquid chromatography. The purified material was analyzed by gas chromatography-mass spectrometry, secondary ion mass spectrometry, and nuclear magnetic resonance spectroscopy. These procedures revealed that this material was 6 beta hydroxydeoxycorticosterone. PMID- 7482629 TI - Inhibition of rat steroid 5 alpha-reductase (isozyme 1) by suramin. AB - In this study, we show the inhibition of rat steroid 5 alpha-reductase (isozyme 1) by suramin. The enzyme activity decreased in a dose-dependent manner as suramin concentrations increased with the calculated drug dose required for 50% inhibition (at 5 microM testosterone and 200 microM NADPH) being 13 microM. Suramin showed non-competitive inhibition of 5 alpha-reductase with respect to testosterone (KT1 = 2.4 microM) and competitive inhibition with respect to NADPH (KiNADPH = 220 nM). Furthermore, suramin and NADP+, but not NAD+, protected 5 alpha-reductase from labeling by 2-azido-NADP+, a photoactive probe which has recently been used to identify the NADPH binding domain of 5 alpha-reductase. These results suggest that suramin inhibits rat steroid 5 alpha-reductase (isozyme 1) at the level of NADPH binding to the enzyme. PMID- 7482631 TI - Inhibition of dexamethasone binding to human glucocorticoid receptor by New World primate cell extracts. AB - To determine if New World primates express an inhibitor that influences glucocorticoid receptor (GR) binding characteristics, we examined [3H]dexamethasone binding in cytosol prepared from B95-8 lymphoid cells, derived from the cotton top tamarin (Saguinus oedipus), in combination with cytosol prepared from human or rat tissues. B95-8 cytosol inhibited specific binding of [3H]dexamethasone (P < 0.01) when mixed with cytosol prepared from either a human lymphoid cell line (HL) or rat thymus. The inhibitory activity was heat labile and trypsin sensitive. Peak inhibitory activity was found in the 150-200 kd fractions after Sephacryl G-200 ultrafiltration. Scatchard analysis of [3H]dexamethasone binding using mixed cytosol showed a diminished GR apparent binding affinity when compared to HL cytosol. Kinetic studies using mixed cytosol indicated that B95-8 cytosol did not affect the apparent dissociation rate of [3H]dexamethasone. These data demonstrate that B95-8 cells contain a competitive inhibitor that prevents binding of dexamethasone to its cognate receptor. PMID- 7482632 TI - Identification of structurally altered estrogen receptors in human breast cancer by site-directed monoclonal antibodies. AB - We have developed and characterized site-directed monoclonal (MAb) and polyclonal antibodies to a specific domain in the N-terminal A/B region in order to assess estrogen receptor (ER) structural integrity in human breast tumor samples. The antibodies (Abs) reacted specifically with the native (undenatured) ER from various species. The synthetic peptides competed effectively for ER binding to the Abs, suggesting site-specificity. The Abs recognized the activated (4S) and transformed (5S) but not the unactivated, untransformed, molybdate-stabilized (8S) ER, suggesting that the epitope is inaccessible in the 8S form. Some of these Abs reacted with ER bound to its responsive elements, as determined by gel mobility shift assay. To evaluate the structural integrity of ER in breast cancer, we have utilized a) ligand binding analysis for the hormone binding domain; b) site-directed MAb to the DNA-binding domain; and c) site-directed MAb to the N-terminal transactivation domain. Analysis of ER from 29 human breast tumors revealed that 10 out of 29 tumors (35%) contained ER with intact hormone-, DNA-, and N-terminal domains. Thirteen out of 29 tumors (approximately 45%) contained ER with intact hormone binding and N-terminal domains but were defective only in the DNA-binding domain. Three out of 29 tumors (approximately 10%) contained ER defective only in the N-terminal domain. Another subgroup of tumors (3/29; approximately 10%) had ER with normal hormone binding domain but were defective in both the DNA-binding and the N-terminal activation domains.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7482633 TI - Enzyme inhibition during the conversion of squalene to cholesterol. AB - Two separate enzymatic assays were developed in order to test the selectivity of inhibitors in cholesterol biosynthesis. One assay detects inhibition of delta 5.7 sterol delta 7-reductase, the enzyme involved in the conversion of 7 dehydrocholesterol to cholesterol. Delta 5.7-Sterol delta 7-reductase was inhibited by both RPR 101821, a protonated cyclohexylamine, and BM 15.766, a piperazine derivative, with IC50 values of 1 microM. The second assay detects accumulation of any of five intermediates (squalene oxide, squalene dioxide, lanosterol, desmosterol, and 7-dehydrocholesterol) upon inhibition of enzymes catalyzing reactions in the conversion of squalene to cholesterol. In this assay, inhibition data were most accurate when control assays exhibited a conversion of squalene to cholesterol in the order of 50%. The time required to attain 50% conversion of squalene to cholesterol was 6 h. Given a high inhibitor to substrate concentration ratio and the possible values of Ki, kon, and koff for the reaction between enzymes and inhibitor to form enzyme-inhibitor complexes, it was predicted that in the presence of inhibitors, intermediate accumulation could still be observed after 6 h incubation. The experimental results were in agreement with this prediction. PMID- 7482634 TI - Estrogen receptor mRNA is expressed in vivo in rat calvarial periosteum. AB - Estrogen deficiency is well recognized as a cause of bone loss in rats and humans. Likewise, treatment with estrogen results in prevention of this loss. Initially, this effect was thought to be indirectly mediated but, more recently, estrogen receptors (ER) have been reported in osteosarcoma cells and primary cultures originating from surgical waste, suggesting a direct effect of this steroid hormone. Detection of ER in skeletal tissues, however, has remained elusive. The purpose of this investigation was to establish the efficacy of the highly sensitive reverse-transcription polymerase chain reaction (RT-PCR) technique to detect ER in a well defined skeletal tissue (calvarial periosteum) that is responsive to the hormone. Primers were made specific to rat ER sequences. Total RNA was extracted from rat uterus, liver, spleen, and the periosteum using an organic solvent method. cDNA was synthesized from 2 micrograms total RNA. cDNA corresponding to 40 ng total RNA/sample produced intense PCR products for ER. In descending order of intensity were uterus, liver, bone, and spleen. Importantly, a similar time-course for estrogen-induced down regulation of steady-state mRNA levels for alkaline phosphatase and osteonectin was observed in calvarial periosteum and tissues known to express estrogen receptors. These data provide in vivo evidence of ER mRNA in bone and suggest that at least some of estrogen's action on bone is directly modulated. PMID- 7482636 TI - Stroke in China, 1986 through 1990. AB - BACKGROUND AND PURPOSE: Incidence of stroke varies markedly in different world populations. In seven Chinese cities, the effect of a program of risk factor modification on the incidence and mortality of stroke was studied and compared with a control population. This article describes the incidence of stroke in the control populations for the years 1986 through 1990. METHODS: Incidence (first ever strokes only) for 1986 was obtained by door-to-door interview with heads of households with subsequent verification on examination by a neurologist and review of medical and/or hospital records. In subsequent years, cases were ascertained with a three-tier monitoring system: by community health workers, local medical centers, and the Beijing Neurosurgical Institute. RESULTS: Average annual age-adjusted incidence per 100,000 was 215.6 (261.5 for males, 174.5 for females; P < .001). There was a significant drop in the total number of cases from 137 in 1986 to 106 in 1990, but the age-adjusted rate showed a significant drop for males only (322.3 to 182.5, P < .001). Marked differences in average annual age-adjusted rates existed among the seven cities, from 486.4 for Harbin to 80.9 for Shanghai. This difference in rate among cities was found for both sexes but was more pronounced in males. CONCLUSIONS: The stroke incidence rates in China, like those in Japan, are among the higher ones in the world. In recent years, there has been an apparent decline in stroke incidence. Marked differences in rates were found between males and females with decline in incidence occurring almost exclusively in males. There were also marked differences in stroke incidence among the cities studied. These differences may result in part from differences in diet, alcohol and cigarette consumption, or prevalence of hypertension. PMID- 7482635 TI - A model to predict the histopathology of human stroke using diffusion and T2 weighted magnetic resonance imaging. AB - BACKGROUND AND PURPOSE: We sought to identify MRI measures that have high probability in a short acquisition time to predict, at early time points after onset of ischemia, the eventual development of cerebral infarction in clinical patients who suffer occlusion of a cerebral artery. METHODS: We developed an MR tissue signature model based on experimentally derived relationships of the apparent diffusion coefficient of water (ADCw) and T2 to ischemic brain tissue histopathology. In eight stroke patients we measured ADCw and T2 intensity using diffusion-weighted echo-planar imaging (DW-EPI). Tissue signature regions were defined, and theme maps of the ischemic focus at subacute time points after stroke onset were generated. RESULTS: Five MR signatures were identified in human stroke foci: two that may predict either cell recovery or progression to necrosis, one that may mark the transition to cell necrosis, and two that may be markers of established cell necrosis. CONCLUSIONS: An MR tissue signature model of ischemic histopathology using ADCw and T2 can now be tested for its potential to predict reversible and identify irreversible cellular damage in human ischemic brain regions. PMID- 7482637 TI - Stroke in young black patients. Risk factors, subtypes, and prognosis. AB - BACKGROUND AND PURPOSE: Stroke subtypes and prognosis differ among older black patients compared with whites; however, few data are available regarding stroke among young black patients. METHODS: To determine the risk factors for stroke, stroke subtype, and prognosis among young black patients, we retrospectively reviewed the medical records of all 15- to 44-year-old patients admitted with stroke to a university-affiliated public hospital from January 1990 through June 1994. RESULTS: Of the 248 eligible patients admitted with stroke, 219 were blacks. Hypertension was more frequently associated with stroke in young black than in non-black patients (55% versus 24%, P = .003). Cocaine abuse was frequent among both black and non-black patients (27% versus 38%, P = NS). Hypertensive intracerebral hemorrhage (64%) was the most common subtype of intracerebral hemorrhage (n = 67), and lacunar infarction (21%) was the most common subtype of cerebral infarction (n = 112) in young black patients. Outcome in black patients with stroke at discharge was 69% independent, 8% dependent, and 23% dead. CONCLUSIONS: The high frequency of hypertension, hypertensive intracerebral hemorrhage, and lacunar infarction among young black patients with stroke suggests accelerated hypertensive arteriolar damage, possibly due to poor control of hypertension. PMID- 7482638 TI - Geographic variation in reporting of stroke deaths to underlying or contributing causes in the United States. AB - BACKGROUND AND PURPOSE: This study examines the geographic variation in the reporting of deaths with stroke as the underlying or contributing cause in the United States. METHODS: Data from the National Center for Health Statistics and Bureau of the Census were used to map the geographic distribution of race- and race/sex-specific, underlying-, contributing-, and multiple-cause age-adjusted stroke mortality rates in the United States by state for 1979 through 1981. RESULTS: Underlying-, contributing-, and multiple-cause age-adjusted stroke mortality rates were significantly clustered for both whites and blacks. However, the spatial distributions of underlying- and contributing-cause rates differed; there was no association between underlying- and contributing-cause rates for either racial group or for the various race/sex groups. There was no association between nonstroke mortality and stroke mortality rates. There was also very little spatial variation and no spatial clustering of the median number of contributing causes reported. CONCLUSIONS: The overall large-scale spatial distribution of resident underlying-cause stroke mortality rates cannot be explained by geographic variation in the selection of the underlying cause of death from among all causes reported on the death certificate, by different area dependent tendencies for mortality generally, or by different tendencies to consider stroke as the cause of death when death occurs. Geographic variation in contributing-cause rates is not explained by variation in tendency to report contributing causes of death. PMID- 7482639 TI - Respiratory function and risk of stroke. AB - BACKGROUND AND PURPOSE: This report examines the relationship between lung function and risk of major stroke events (fatal and nonfatal). METHODS: We completed a prospective study of 7735 men aged 40 to 59 years at screening selected at random from one general practice in each of 24 British towns. RESULTS: During the mean follow-up period of 14.8 years, there were 277 major stroke events in the 7650 men with data on forced expiratory volume in 1 second (FEV1). After exclusion of 499 men with definite myocardial infarction, stroke, or atrial fibrillation at screening, 7151 men experienced 239 major stroke events. Lower levels of FEV1 were associated with a significant increase in risk of stroke even after adjustment for age, smoking, social class, physical activity, alcohol intake, systolic blood pressure, antihypertensive treatment, diabetes, and preexisting ischemic heart disease. Relative risk in the low third (< 3.10 L) versus high third (> 3.65 L) was 1.4 (95% confidence interval, 1.0 to 2.0). The inverse association between FEV1 and stroke was only apparent in older men, current nonsmokers, hypertensive men, and men with preexisting ischemic heart disease. Lower FEV1 was associated with higher rates of stroke in hypertensive men irrespective of smoking status. Inclusion of FEV1 in a risk score for stroke provided only a small increase in the absolute risk or the yield of cases in the top fifth of the score distribution during the follow-up period. CONCLUSIONS: Lower levels of FEV1 are associated with an increased risk of stroke in those already at high risk, eg, those with ischemic heart disease or hypertension. However, the association is not strong enough to warrant the use of FEV1 in making clinical decisions regarding the treatment of hypertension as it relates to the prevention of stroke. PMID- 7482641 TI - Influence of vascular risk factors for atherosclerotic carotid artery plaque progression. AB - BACKGROUND AND PURPOSE: Investigations regarding arteriosclerosis of carotid arteries showed an association between increased intima-media thickness and vascular risk factors. A newly developed three-dimensional ultrasound method increases the reproducibility of plaque volume measurements because more exact volume measurements can be performed with a reduction of the disadvantages of two dimensional measurements. In a pilot study the influence of vascular risk factors on carotid artery plaque progression was examined. METHODS: Volumes of atherosclerotic plaques in carotid arteries in 54 patients were measured with a three-dimensional ultrasound system during a 12-month period to determine the relationship between progression or regression of plaque volume, vascular risk factors, dose of aspirin, and flow turbulence in the plaque region. RESULTS: A progression of plaque volume occurred in 67% (36/54) of all plaques. In no plaque was a regression of plaque volume seen. The optimal adjustment of all risk factors showed a significant influence on plaque progression (r = .31). Diastolic blood pressure was the strongest predictor of plaque progression (P < .01), followed by diabetes (P < .03). Turbulence in the plaque region was found in 78% of the patients in the progression group (n = 36) versus 61% in the nonprogression group (n = 18) but was not significant. Dose of aspirin (100 mg versus 250/300 mg) had no influence on plaque volume after 1 year. CONCLUSIONS: Treatment of vascular risk factors reduces the progression of carotid artery plaque volume in three-dimensional ultrasound. The most important factor for plaque progression is a high diastolic blood pressure. Turbulence in the flow pattern and the examined doses of aspirin showed no significant influence. PMID- 7482640 TI - Prognosis for survival after an initial stroke. AB - BACKGROUND AND PURPOSE: We studied prognosis for survival after an initial stroke in 662 patients who survived at least 30 days after onset while taking into account age, sex, the number of neurological deficits from the initial stroke, stroke type, and five selected medical conditions: hypertension, myocardial infarction, cardiac arrhythmia, diabetes mellitus, and history of transient ischemic attacks. METHODS: All patients were enrolled between July 1, 1987, and August 1, 1989, and were followed regularly at about 6-month intervals until death or the end of the study (mean of 24 months). RESULTS: At 6 months, 90.8% of the 30-day stroke survivors were still alive. At 1, 2, 3, and 4 years, the cumulative survival rates were 86.9%, 78.7%, 73.2%, and 72.0%, respectively. Older age and the number of neurological deficits at onset of initial stroke increased risk of death. Compared with patients of the same age, sex, number of neurological deficits, and comorbidities, increased risk of death is present among those with myocardial infarction, cardiac arrhythmia, and diabetes mellitus. Hazard ratios were 1.7 (P = .006), 1.5 (P = .023), and 1.4 (P = .059), respectively. Hypertension and transient ischemic attacks were not significantly associated with increased mortality. CONCLUSIONS: This study clarifies prognosis for survival after an initial stroke by taking into account other confounding variables that could also contribute to risk of death. PMID- 7482642 TI - Functional outcome for patients with hemiparesis, hemihypesthesia, and hemianopsia. Does lesion location matter? AB - BACKGROUND AND PURPOSE: Patients with hemiparesis, hemisensory loss, and hemianopsia ("HHH" deficits) due to stroke may have large cortical lesions caused by middle cerebral trunk vessel occlusion or smaller subcortical lesions due to lenticulostriate involvement. We studied the usefulness of lesion location in predicting functional recovery within this syndrome. METHODS: We reviewed our records and found 41 patients who had a single ischemic hemispheric stroke, HHH deficits, and an available CT scan performed more than 24 hours after the onset of symptoms. CT scans were read independently and blindly by the authors. Lesions were initially categorized by arterial distribution on the basis of CT templates published by Kinkel. The numerous combinations of arterial branch vessel occlusions observed did not allow for statistical analyses because of the small number of subjects within each subgroup. Lesions were therefore classified as cortical (C), subcortical (S), or mixed (M). RESULTS: There were no significant differences among the three anatomic groups for age, sex, interval after stroke, Mini-Mental Status Examination score, or admission Barthel Index score. Functional outcome measures did not differ significantly for the three groups: mean +/- SD discharge Barthel score (C, 64 +/- 31; S, 47 +/- 20; M, 57 +/- 21), length of stay ([days] C, 64 +/- 25; S, 77 +/- 24; M, 73 +/- 28), and frequency of nursing home placement (C, 4/8; S, 3/6; M, 2/16). CONCLUSIONS: For patients with HHH deficits, the anatomic location of the lesion (C versus S versus M) does not affect functional outcome. PMID- 7482643 TI - The clinical meaning of Rankin 'handicap' grades after stroke. AB - BACKGROUND AND PURPOSE: The Rankin Scale is a frequently used handicap index in stroke outcome research. However, relatively little is known about its validity. The purpose of this study was to investigate the clinical meaning of Rankin grades by identifying the functional health aspects that contribute to Rankin scores. METHODS: We studied 438 patients 6 months after stroke. Data were collected on the following functional health indicators: alertness, communication, independence, disability in activities of daily living, mobility, instrumental disability, social interaction, and recreation. Disability in activities of daily living was assessed with the Barthel Index, whereas the other indicators were measured with subscales of the Sickness Impact Profile. The association between functional health and Rankin Scale was expressed in terms of relative frequencies and Somers' D statistic. Linear regression analysis (after ordinal transformation) was used to identify the significant health factors that explain Rankin scores. RESULTS: Mobility, disability in daily and instrumental activities, and living arrangements showed a stronger association with Rankin scores (Somers' D range, 0.60 to 0.74) than cognitive and social functioning (Somers' D range, 0.34 to 0.47). Disability in activities of daily living turned out to be the most important explanatory factor of Rankin scores (R2 = 67%). CONCLUSIONS: The Rankin Scale is not a pure handicap measure but should be viewed as a global functional health index with a strong accent on physical disability. The index is useful as a simple and time-efficient outcome measure in largescale multicenter trials. It is argued that at present there is no clear need to assess handicap as the primary outcome in medically oriented stroke intervention studies. PMID- 7482644 TI - Role of stroke rehabilitation units in managing severe disability after stroke. AB - BACKGROUND AND PURPOSE: Stroke unit rehabilitation tends to be directed toward stroke patients with moderately severe disabilities ("the middle group"). Data collected on a stroke rehabilitation unit, however, showed improving outcome over 3 years in patients with a poor prognosis (discharge home: 48% versus 16%, P < .02; discharge Barthel Index score: 9 versus 6, P < .05). The hypothesis that stroke rehabilitation units may improve outcome in severely disabled stroke patients was tested in this study. METHODS: A randomized controlled study was undertaken in 71 patients with a poor prognosis who were treated either on a stroke rehabilitation unit (n = 34) or on general wards (n = 37) to compare outcome between the two groups. Data collected were also compared with those from a methodologically similar study undertaken 3 years ago. RESULTS: Severe stroke patients treated on the stroke rehabilitation unit had a significantly better outcome compared with general wards (mortality: 21% versus 46%, P < .05; discharge home 47% versus 19%, P < .01; median length of hospital stay: 43 versus 59 days, P < .02). The number of stroke unit patients being discharged home had increased significantly from the previous study, with a trend toward improvement in median discharge Barthel Index score. CONCLUSIONS: Stroke rehabilitation units may improve outcome in severe stroke patients. This improvement appears to be due to the development of innovative management strategies that reduce mortality and institutionalization and enable caregivers to support more disabled stroke patients at home. PMID- 7482645 TI - Amino acid transmitters in patients with headache during the acute phase of cerebrovascular ischemic disease. AB - BACKGROUND AND PURPOSE: The pathophysiology of headache occurring at stroke onset is unknown. Migraine and ischemia share an excessive release of neuroexcitatory amino acids. Inhibitory amino acids also may be implicated in both diseases. We investigated whether fluctuations of these amino acids occur in headache accompanying cerebral infarction. METHODS: We studied 100 patients with infarction in the territory of the middle cerebral artery. Neurological impairment was assessed using the Canadian Neurological Scale and Barthel Index. Size of infarction was determined with CT. Twenty-eight patients developed headache. Glutamate, aspartate, and taurine were quantified in blood and cerebrospinal fluid (CSF) within 24 hours of stroke onset with cationic exchange chromatography. RESULTS: Stroke subtypes, size of infarct on CT, and clinical scales were similar in patients with and without headache. Plasma glutamate level was 321.14 +/- 149.53 mumol/L in patients with headache and 233 +/- 107.23 mumol/L in those without headache (P < .005). Glutamate in CSF was higher in patients with headache (4.6 +/- 1.49 mumol/L) than in patients without headache (3.11 +/- 1.18 mumol/L) (P < .001). Aspartate concentrations in plasma and CSF were similar in both groups. Taurine concentrations in plasma were 103.10 +/- 52.82 mumol/L and 177.49 +/- 90.92 mumol/L in headache and nonheadache patients, respectively (P < .001). Taurine levels in CSF were 5.42 +/- 2.42 mumol/L in patients with headache and 9.27 +/- 5.31 mumol/L in those without headache (P < .001). No significant correlation was found between amino acid levels in plasma or CSF and size of infarction. CONCLUSIONS: Amino acid neurotransmitters play a role in the pathophysiology of headache that occurs at the onset of stroke. The ischemic penumbral area, more than the infarction itself, may cause a state of cortical hyperexcitability that would be responsible for the cortical release of amino acids and the induction of headache by altering pain perception mechanisms. PMID- 7482646 TI - Fever in acute stroke worsens prognosis. A prospective study. AB - BACKGROUND AND PURPOSE: No definitive data are yet available on the effects of body temperature on neurological damage after cerebral ischemia in humans. Experimental animal models have provided much evidence, but to our knowledge, only two studies on the relationship between fever and prognosis of stroke in humans have been published. The aim of our study was to investigate the prognostic role of fever in the first 7 days of hospitalization in a cohort of patients admitted to our hospital for acute stroke. METHODS: We analyzed the data of 183 patients included in a prospective observational prognostic study. Vital status at 30 days was considered the main outcome and was obtained for all patients. Age, level of consciousness, and glycemia at the time of hospitalization were considered covariates for an exact logistic regression analysis. The maximum temperature recorded during the first 7 days dichotomized as "no or low fever" versus "high fever" was added to the model. Death within 10 days, taken as a secondary outcome suggestive of death from neurological causes, was analyzed with exact permutation tests. RESULTS: Of the 183 patients analyzed in this study, 43% had fever during the first 7 days after hospitalization. The mean value of the maximum temperature recorded during the first 7 days in the 78 febrile patients was 38.3 degrees C, and the median was 37.9 degrees C. Onset of fever occurred in only 15% of febrile patients during the first day and in 49% on the second. The prognostic roles of age, level of consciousness, and glycemia were confirmed by exact logistic regression. Degree of consciousness impairment was the strongest prognostic variable, with an odds ratio (OR) of 11.4 (95% confidence interval [CI], 4.4 to 31.6). High fever (maximum temperature recorded during the first 7 days > or = 37.9 degrees C) was an independent factor for a worse prognosis, with an OR of 3.4 (95% CI, 1.2 to 9.5). The OR of dying within 10 days versus dying between 11 and 30 days was 4.9 (95% CI, 1.2 to 25.2) in patients with high fever with respect to all other patients. CONCLUSIONS: Fever in the first 7 days was an independent predictor of poor outcome during the first month after a stroke. No data were available on the underlying causes of fever, but the higher risk of death in the first 10 days, most frequently attributed to neurological mechanisms, suggested that high temperature was an independent component of poor prognosis and not only an epiphenomenon of other complications in the course after a stroke. In agreement with animal studies, we found that patients with higher temperature had a worse stroke outcome. PMID- 7482647 TI - Importance of time-window overlap in the detection and analysis of embolic signals. AB - BACKGROUND AND PURPOSE: The detection of asymptomatic embolic signals by Doppler ultrasound may offer a powerful investigational tool in the management of cerebrovascular disease. However, early studies, particularly in patients with carotid artery disease, have reported very different frequencies of embolic signals. While this may reflect differences in patient groups and the criteria used for embolic signal identification, the degree of time-window overlap may be important. If this is insufficient, some embolic signals may fall between two time windows and not appear on the spectral display. Furthermore, the use of nonrectangular time windows, such as the Hanning window, may result in variation of the intensity of an embolic signal depending on where it is detected within the time window. METHODS: To test the importance of this potential problem, the same 25 embolic signals recorded as the audio signal on digital audiotape were each played repeatedly through a transcranial Doppler ultrasound (TCD) system using fast Fourier transform analysis. An older system with no time-window overlap was used, and a more modern system was also used in which three different degrees of overlap were used: -9%, 27%, and 57%. The number of signals audible but not appearing on the spectral display was recorded. The variability in the relative intensity increase for the same embolic signal played repeatedly was estimated by calculating the coefficient of variation of the relative intensity increase. RESULTS: With the older system, 39/500 (7.8%) of embolic signals were missed. With the newer system, the number of embolic signals missed was fewer and decreased with increasing degrees of overlap (10/500 for -9% overlap, 1/500 for 27% overlap, and 0/500 for 57% overlap). For those setups in which embolic signals were missed, there was a highly significant relationship between duration of embolic signal and number of signals missed. In parallel with these results, the coefficient of variation of the relative intensity increase became progressively less with increasing degrees of time-window overlap. For all processing setups, the coefficient of variation was greater for the less intense and shorter duration signals, but this dependence, as estimated by the slope of the regression line, became less strong with higher degrees of overlap. CONCLUSIONS: Inadequate degrees of fast Fourier transform time-window overlap will result in the failure of current TCD machines to detect embolic signals. Furthermore, this and the time windowing currently usually used may result in variability in the relative intensity increase of identical embolic signals. These factors need to be taken into account when comparing data on the frequencies of embolic signals recorded by different researchers and in the design of future TCD equipment. PMID- 7482648 TI - Phase I: transcranial echo contrast studies in healthy volunteers. AB - BACKGROUND AND PURPOSE: Transcranial ultrasound diagnostics are particularly hindered by insufficient ultrasound penetration through the temporal bone. The use of ultrasonic contrast media to enhance the Doppler signal is an important step toward the solution of this problem. In the present study we investigated the tolerability and the diagnostic value of a new intravenous transpulmonary ultrasonic contrast medium, BY963. METHODS: In two phase I studies, 8 healthy volunteers received a spherosome suspension containing a phospholipid as the active ingredient. The intravenous injection was performed in three doses (2.5, 5, and 10 mL) at four different injection rates (0.25, 0.5, and 1 mL/s and bolus). The duration and degree of the signal enhancement were measured by two transcranial ultrasonic procedures presently used in clinical practice: transcranial Doppler sonography (TCD) and transcranial color-coded sonography (TCCS). The assessment of tolerability was based on chemical laboratory parameters and hemodynamic data (heart rate, blood pressure, electrocardiogram) and on questionnaires relating to general well-being. RESULTS: BY963 was tolerated without complications. All 38 administrations of the echo contrast medium produced a marked increase in the TCD signal (> 30 dB) in the intracranial basal cerebral arteries. To obtain the optimum time window for diagnostic use, higher doses with slower injection rates are advantageous. The duration of optimal contrasting was 42 to 68 seconds (TCD) and 12 to 132 seconds (TCCS), depending on the method and mode of administration. Bolus injections gave rise to an increased incidence of color artifacts. CONCLUSIONS: BY963 significantly improves intracranial Doppler imaging while being well tolerated. The signal enhancement lasts long enough for TCCS to display all basal cerebral arteries after just one injection. PMID- 7482649 TI - Can transcranial Doppler really detect reduced cerebral perfusion states? AB - BACKGROUND AND PURPOSE: This study was designed to determine whether transcranial Doppler ultrasonography (TCD) may detect reduced perfusion states of the brain in patients with hypertension or diabetes mellitus with suspected cerebral atherosclerosis and arteriolosclerosis. METHODS: We determined blood flow velocity with TCD in the middle cerebral artery and cerebrovascular vasodilator responses to carbon dioxide in 22 patients with or without carotid artery occlusive disease and minor stroke; we compared the results with the measurements of cerebral blood flow and oxygen metabolism by positron emission tomography (PET). RESULTS: Blood flow velocity measured by TCD correlated with ipsilateral cerebral blood flow measured by PET in frontal, temporal, and striatal regions and throughout the entire hemisphere (P < .05 to P < .005). Relative changes in blood flow velocity and calculated cerebrovascular resistance tested by carbon dioxide inhalation both correlated closely with regional mean transit time (calculated as the ratio of cerebral blood volume divided by cerebral blood flow) in frontal, striatal, temporal, parietal, and occipital regions and also in the entire hemisphere (P < .05 to P < .0001). TCD variables did not correlate with hemispheric measurements of oxygen metabolism by PET. CONCLUSIONS: Although TCD is not useful in assessing impairments of cerebral metabolism, it is useful for detecting abnormalities of cerebral hemodynamics among patients with risk factors for cerebrovascular disease. PMID- 7482650 TI - Potential and limitations of transcranial color-coded sonography in stroke patients. AB - BACKGROUND AND PURPOSE: Transcranial color-coded duplex sonography (TCCS) enables visualization of the intracranial parenchymal structures and measurement of blood flow velocity in the basal cerebral arteries. The present study aims to evaluate prospectively the clinical usefulness of TCCS in patients with acute stroke. METHODS: Eighty-four consecutive patients with central nervous symptoms suggesting acute stroke were investigated within the first 48 hours after clinical onset. TCCS was performed with a 2.5-MHz sector transducer through the temporal bone window. CT was available in all patients. RESULTS: Forty-eight patients suffered from an infarction or a transient ischemic attack (TIA) in the territory of the middle cerebral artery (MCA). Fifteen of them showed an MCA occlusion, and 12 of the 15 developed recanalization during follow-up. Twelve revealed an increased, decreased, or oscillating flow pattern in the MCA main stem, and 21 patients had no ultrasonic abnormalities. The positive and negative predictive values of a pathological flow pattern in patients with MCA infarctions or TIA were .92 and .48, respectively. Fifteen patients suffered from an intracerebral hematoma, which could be diagnosed by TCCS in 14 cases. The positive and negative predictive values of a pathological parenchymal echo pattern were .88 and .96, respectively. Three patients suffered from an infarction and one from a TIA in the posterior cerebral artery territory. One female patient with an acute deterioration of a hemiparesis showed a glioma. The dropout rate due to an insufficient acoustic temporal bone window was 20% (17/84). CONCLUSIONS: TCCS is a noninvasive bedside method that provides rapid and reliable data regarding stroke subtype and mechanism immediately after onset. Window failure is a serious limitation of this method. PMID- 7482651 TI - Occurrence of transcranial Doppler high-intensity transient signals in patients with potential cardiac sources of embolism. A prospective study. AB - BACKGROUND AND PURPOSE: Cerebral emboli can be recognized by typical "high intensity transient signals" (HITS) in the transcranial Doppler (TCD) spectral curves. Patients with potential cardiac sources of embolism are at higher risk for stroke. METHODS: We examined the frequency of HITS in the left middle cerebral artery (MCA) with TCD over periods of 30 minutes in 100 patients having potential cardiac sources of embolism, as indicated by transthoracic or transesophageal echocardiography. RESULTS: Thirty-six (36%) of the patients presented with HITS. Sex, age, sufficient anticoagulation level, antiplatelet therapy, neurological symptoms, and a history of thrombosis had no influence on the prevalence and number of HITS. The patients with a single echocardiographic diagnosis were separated into eight echocardiographically defined groups: patients with (1) atrial fibrillation, (2) coronary artery disease plus ejection fraction of more than 30% including at least three wall segments of hypokinesia/akinesia, (3) coronary artery disease with less than 30% ejection fraction, (4) dilated cardiomyopathy, (5) infectious endocarditis, (6) aortic stenosis, (7) mitral stenosis, and (8) patent foramen ovale. A significant difference in HITS occurrence could not be found in any of the defined groups. Only patients with infectious endocarditis showed a tendency for a higher HITS prevalence. CONCLUSIONS: HITS are common phenomena in patients with potential cardiac sources of embolism. The clinical relevance of these HITS remains unclear. PMID- 7482652 TI - Transcranial Doppler ultrasound in hypertensive versus normotensive patients after aneurysmal subarachnoid hemorrhage. AB - BACKGROUND AND PURPOSE: Arterial hypertension is a negative prognostic risk factor after aneurysmal subarachnoid hemorrhage (SAH). Transcranial Doppler ultrasound is commonly used for measuring blood flow velocities to predict cerebral ischemia due to vasospasm after SAH. Our purpose was to evaluate the influence of arterial hypertension on blood flow velocities in the cerebral circulation after aneurysmal SAH. METHODS: With transcranial Doppler ultrasound we compared the blood flow velocities in matched groups of hypertensive and normotensive patients with aneurysmal SAH. Twenty-four patients with arterial hypertension were examined daily during a 2-week period. As controls, 24 normotensive patients, also with SAH, were matched by age, sex, neurological status, and clinical outcome. RESULTS: Minimum, mean, and maximum flow velocities in the middle cerebral artery in the hypertensive patients were significantly lower than in the normotensive individuals, with P = .02 for minimum, P = .02 for mean, and P = .02 for maximum. There was no statistical significance for pulsatility index differences in these groups (P = .45). Diastolic notch was noted in two of the hypertensive and in six of the normotensive patients. CONCLUSIONS: The results indicate that even moderately increased flow velocities in hypertensive patients may represent significant vasospasm. PMID- 7482654 TI - Induction of cerebral thrombosis with phenytoin in rats. AB - BACKGROUND AND PURPOSE: This study was conducted to explore whether intra arterial infusion of phenytoin causes cerebral ischemia and to examine the mechanism of cerebral ischemia induced by phenytoin. METHODS: Ten rats were infused with phenytoin (150 microL, 3.75 mg) retrogradely from the left external carotid artery, followed by perfusion of carbon black transcardially. The removed brain was photographed from above, and the nonperfused area was compared with control rats (n = 10) with the use of an image analyzer. Eight animals with or without phenytoin treatment were perfusion-fixed for transmission electron microscopic analyses of cerebral vasculature. To determine the effect of tissue plasminogen activator (TPA) on phenytoin-infused rat cerebrum, 20 rats were treated with or without TPA (120,000 IU) 5 minutes after the phenytoin infusion (n = 10 each). RESULTS: All rats suffered from respiratory distress 25 to 40 minutes after the injection and received carbon black transcardially. The nonperfused area was seen in the territory of the left internal carotid artery. Thrombi were observed from arterioles to capillaries. Under electron microscopy, endothelial cells were partially exfoliated, and the vascular lumen was obstructed by thrombi predominantly consisting of platelets. Eight rats with TPA survived more than 60 minutes, whereas only 2 rats survived without the treatment (P < .005). Nonperfused areas were 7 +/- 5% and 50 +/- 11% of cerebral surface area in rats with and without TPA treatment, respectively (P < .001). CONCLUSIONS: Intra-arterial infusion of phenytoin results in a nonperfused area in rat cerebrum primarily due to thrombosis of arterioles and capillaries. PMID- 7482653 TI - Effect of Guglielmi detachable coils on experimental carotid artery aneurysms in primates. AB - BACKGROUND AND PURPOSE: Clinical experience has established that intravascularly placed metal coils can be a useful treatment for cerebral vascular aneurysms. However, the mechanism by which the coils induce occlusion of the aneurysm is unclear. Appropriate use of this promising treatment modality requires basic understanding of the occlusive process. We used an animal model system of experimentally induced carotid aneurysms to investigate the initial events induced by Guglielmi detachable coils (GDCs), as well as the subsequent vascular changes induced by the coils over time. METHODS: We induced 23 aneurysms in the carotid arteries of 16 Japanese monkeys. Nineteen aneurysms were then occluded with GDCs placed via endovascular surgery; 4 aneurysms served as controls. We then used gross and microscopic pathological examination, angiography, and scanning electron microscopy to assess the effects of the GDC. RESULTS: In the first few hours after placement of the GDC in the experimental aneurysms, we observed leukocyte attachment and deposition of fibrinlike materials and other proteins. By 4 days after coil placement, leukocytes and fibroblasts were observed in the thrombus. By 2 weeks after coil placement, there was evidence of an endothelial-like covering of the coils. At 3 months after coil placement, we observed development of an arterial media in the occluded aneurysms. CONCLUSIONS: The GDCs initiated a cellular response within several hours of aneurysm occlusion. By 2 weeks after coil placement, endothelialization was proceeding, and by 3 months after occlusion, remodeling of the aneurysm had progressed to produce a media-like structure in the former aneurysm. PMID- 7482655 TI - A reproducible model of middle cerebral infarcts, compatible with long-term survival, in aged rats. AB - BACKGROUND AND PURPOSE: Stroke is a disease associated with aging, but experimental stroke studies are generally done in young male animals. Because there are numerous differences associated with aging, such as an altered immune system and altered neurochemistry, that could affect the outcome of these experiments, a model of reproducible cerebral infarction in aged rats is needed. METHODS: We attempted to produce middle cerebral artery (MCA) infarcts in aged (22 months of age) rats using two standard methods. A nylon suture with a heat induced bulb was passed through the external carotid artery in seven animals, with an attempt to place the tip at the origin of the MCA. The MCA was ligated through a craniotomy just proximal to the internal cerebral vein in 14 rats. Survival potential was tested by attempting 2-week survival in four rats and 2 month survival in one rat. RESULTS: The suture model failed to produce MCA infarcts, even when the bulb of the suture was properly placed in the MCA. The intracranial MCA occlusion resulted in reproducible MCA infarcts. There were no deaths, including the animals allowed to survive 2 weeks and 2 months. CONCLUSIONS: We conclude that reproducible MCA infarcts can be produced in aged rats by craniotomy and that these lesions may be compatible with long-term survival. This should be a useful technique for studying therapeutic interventions and rehabilitation strategies in an animal model that immunologically and neurochemically more closely resembles humans at risk for stroke. PMID- 7482656 TI - Global incomplete cerebral ischemia produces predominantly cortical neuronal injury. AB - BACKGROUND AND PURPOSE: We determined the neuropathologic damage in a canine model of global incomplete ischemia commonly used in a variety of physiological experiments. METHODS: We induced 20 minutes of incomplete ischemia in dogs (n = 9) by increasing intracranial pressure via intraventricular infusion of artificial cerebrospinal fluid to maintain a cerebral perfusion pressure of 10 mm Hg while keeping body temperature at 38 degrees C during and immediately after ischemia. After a 7-day recovery period, animals were perfusion-fixed for neuropathology. In hematoxylin and eosin preparations, ischemic neuronal injury was assessed, neurons were counted, and percentage of cell damage was determined. RESULTS: No focal neurological deficits or overt seizures were observed during the 7-day recovery period. In superior temporal gyrus, 49 +/- 11% and 70 +/- 10% damage (mean +/- SEM) was observed in layer III pyramidal cells in the crown and sulcus, respectively. All neocortical regions examined showed neuronal damage in layers III and/or V. In hippocampus, 59 +/- 11% damage of pyramidal neurons occurred in CA1, with dorsal (septal) hippocampus showing more injury than ventral (temporal) portions. The caudate nucleus (head) exhibited 27 +/- 7% neuronal injury. In cerebellar cortex (anterior lobule), 70 +/- 7% damage of Purkinje cells occurred, but different folia of cerebellum showed varying degrees of injury. Brain stem and thalamus were minimally affected despite reduced blood flow. Inflammatory changes (leukocytic infiltration and neuronal incrustations) were observed, but only when neuronal degeneration was severe. Pancellular necrosis and infarction did not occur. CONCLUSIONS: This animal model of ischemia causes reproducible neuronal injury primarily in cortical regions without pancellular necrosis and infarction. Damage to subcortical areas is less severe than to cortical areas, despite comparable reductions in regional cerebral blood flow. Therefore, in the presence of regionally uniform but incomplete cerebral ischemia, neocortical and hippocampal pyramidal neurons and cerebellar cortical Purkinje cells are more likely than subcortical neurons to degenerate; alternatively, pyramidal and Purkinje neurons degenerate before neostriatal neurons in this model. This neuronal degeneration may represent an intrinsic cellular mechanism without major contribution of cytotoxic pathways associated with inflammation. PMID- 7482657 TI - Acute effects of ethanol on responses of cerebral arterioles. AB - BACKGROUND AND PURPOSE: Previous studies have suggested that acute exposure of large peripheral arteries to ethanol impairs endothelium-dependent relaxation. The goal of the present study was to determine the acute effects of ethanol exposure on responses of cerebral resistance arterioles in vivo. METHODS: We prepared a cranial window in rats to expose the cerebral (pial) microcirculation. We measured the diameter of pial arterioles in vivo in response to agonists that presumably stimulate the synthesis/release of nitric oxide from the endothelium (ADP, acetylcholine, and histamine) or neurons (N-methyl-D-aspartate [NMDA]) before and after topical application of various concentrations of ethanol added to the cerebrospinal fluid (20, 40, 60, 80, and 100 mmol/L). In addition, we examined responses of pial arterioles to nitroglycerin before and 1 hour after topical application of ethanol. RESULTS: Before application of ethanol, ADP, acetylcholine, histamine, NMDA, and nitroglycerin produced dose-related dilatation of pial arterioles. Application of the various concentrations of ethanol did not alter the baseline diameter of pial arterioles. However, application of 80 and 100 mmol/L ethanol inhibited dilatation of pial arterioles in response to agonists that stimulate the synthesis/release of nitric oxide. Dilatation of pial arterioles in response to nitroglycerin was not altered by application of ethanol. CONCLUSIONS: The findings of the present study suggest that acute exposure of cerebral arterioles to modest-to-moderate concentrations of ethanol (20 to 60 mmol/L) does not alter responses of cerebral arterioles. In contrast, exposure of cerebral arterioles to higher concentrations of ethanol (80 and 100 mmol/L) can produce specific impairment of dilatation to agonists that stimulate the synthesis/release of nitric oxide from endothelium and neurons. PMID- 7482658 TI - Prostacyclin receptor activation and pial arteriolar dilation after endothelial injury in piglets. AB - BACKGROUND AND PURPOSE: Both light/dye endothelial injury and indomethacin treatment inhibit hypercapnia-induced cerebral prostacyclin synthesis and pial arteriolar dilation in newborn pigs. Topical iloprost can allow hypercapnia induced dilation of pial arterioles to occur in piglets treated with indomethacin. We addressed the hypothesis that prostacyclin receptor activation with iloprost can return the ability of pial arterioles with endothelial injury to respond to hypercapnia. We also examined whether the endothelial dependence and the permissive role of prostacyclin extended to histamine-induced dilation or are specific for hypercapnia. METHODS: Experiments used chloralose-anesthetized piglets equipped with closed cranial windows. Hypercapnia (PaCO2 approximately 80 mm Hg) and topically applied histamine (10(-6) and 10(-5) mol/L) dilated pial arterioles. Dilations in response to both stimuli were abolished by light/dye treatment. RESULTS: Simultaneous topical treatment with iloprost (10(-12) mol/L, which caused no residual dilation, returned dilation of pial arterioles to both hypercapnia and histamine. On removal of iloprost, responses were again absent and returned with readdition of iloprost to the cortical cerebrospinal fluid. Neither isoproterenol nor sodium nitroprusside returned responses to hypercapnia after light/dye treatment. CONCLUSIONS: These data add further support to the hypothesis that prostacyclin represents an important endothelial-derived signal in the newborn pig cerebral circulation that can permit appropriate responses by adjacent smooth muscle in response to specific stimuli. PMID- 7482659 TI - Sequential studies of severely hypometabolic tissue volumes after permanent middle cerebral artery occlusion. A positron emission tomographic investigation in anesthetized baboons. AB - BACKGROUND AND PURPOSE: In the positron emission tomography literature, markedly hypometabolic brain tissue (oxygen metabolism < 1.3 to 1.7 mL.100 g-1.min-1) has often been equated with irreversible damage in the human brain. By serial positron emission tomography measurements, we investigated the temporal evolution of the volume of severely hypometabolic brain tissue after permanent middle cerebral artery occlusion in anesthetized baboons with, as a perspective, the development of rational therapeutic strategies. METHODS: Seven anesthetized and ventilated baboons underwent sequential positron emission tomography examinations with the 15O steady-state technique before and 1, 4, 7, and 24 hours and 14 to 29 days after occlusion. In each baboon the infarct volume was calculated by quantitative histological procedures after 19 to 41 days of occlusion. RESULTS: The sequential measurement of regional oxygen metabolism demonstrated an extension (for > or = 24 hours) of the volume of severely hypometabolic tissue as defined by both absolute and relative metabolic thresholds, and this profile of evolutivity is observed no matter the threshold used. Mean (+/- SEM) infarction volume of 2.4 +/- 0.6 cm3 was comparable to a tissue volume with oxygen consumption < 40% of contralateral metabolism. The volume of hypometabolic tissue was essentially stable at the 1-, 4-, and 7-hour postocclusion studies, increased markedly at the 24-hour study point, and increased even further in the chronic stage study (on average, 17 days after occlusion). The tissue that eventually displayed a severely hypometabolic state at the final measurement showed a significant decrease of oxygen metabolism and cerebral blood flow at each time analyzed. In that tissue, the oxygen extraction fraction increased significantly at 1 hour (although not thereafter). CONCLUSIONS: The extension of severely hypometabolic volume after middle cerebral artery occlusion reinforces the concept of a dynamic penumbra and suggests the existence of a relatively large window of therapeutic opportunity in which it may be possible to develop neuroprotective strategies. Our study suggests that maximum infarct volume is determined at some time between 24 hours and 17 days after permanent middle cerebral artery occlusion in anesthetized baboons. PMID- 7482660 TI - Microvascular basal lamina antigens disappear during cerebral ischemia and reperfusion. AB - BACKGROUND AND PURPOSE: Changes in vascular permeability are well-known and important consequences of cerebral ischemia. The development of edema and of petechial hemorrhage is connected to altered vascular integrity. A major part in microvascular integrity is played by the basal lamina. METHODS: The fates of the basal lamina components laminin, fibronectin, and type IV collagen during middle cerebral artery occlusion (2 hours, n = 3) and occlusion (3 hours) with reperfusion (1 hour, n = 3; 4 hours, n = 3; and 24 hours, n = 4) were evaluated in the nonhuman primate. Specific monoclonal antibodies against these components were used. The number and size distribution of the microvessels in each specimen were determined by video-imaging microscopy, and the relative fluorescence intensity of laminin was semiquantified by laser confocal microscopy. Basal lamina antigen presentations were compared by double-stain immunofluorescence histochemistry. RESULTS: The number of microvascular structures defined by the presence of each basal lamina antigen decreased significantly up to 24 hours of reperfusion (P < .0001). The ratio of laminin-containing vessels between the ischemic and nonischemic territories decreased significantly from control (0.98 +/- 0.04) to 2 hours of ischemia (0.83 +/- 0.09) and 1 hour (0.79 +/- 0.08), 4 hours (0.77 +/- 0.06), and 24 hours of reperfusion (0.55 +/- 0.07). The ratio of fibronectin (cellular) and of collagen (IV)-containing vessels decreased from 0.98 +/- 0.04 to 0.75 +/- 0.1 and from 1.02 +/- 0.03 to 0.57 +/- 0.1, respectively. Mean laminin fluorescence intensity decreased from 76.1 +/- 6.0 U (controls) to 52.0 +/- 14.6 U (24 hours of reperfusion; P < .001). CONCLUSIONS: The significant parallel losses of three basal lamina components, both in number and intensity, contribute to loss of microvascular integrity. These phenomena may be important for understanding cell extravasation and the hemorrhagic consequences of acute stroke. PMID- 7482661 TI - Regional prevalence and distribution of ischemic neurons in dog brains 96 hours after cardiac arrest of 0 to 20 minutes. AB - BACKGROUND AND PURPOSE: In this established outcome model of cardiac arrest in dogs, we have used total (summed regional) brain histopathologic damage scores. The present study describes the regional progression of necrotic (ischemic) neuron prevalence with increasing duration of cardiac arrest. It tests the hypothesis that increases in the total prevalence of necrotic neurons better correspond to increasing arrest duration and better correlate with neurological deficit than do any individual regional scores. METHODS: Blinded evaluation with light microscopy was used to score the prevalence (five categories) and note the distribution of necrotic neurons in dog brains 96 hours after normothermic ventricular fibrillation cardiac arrest followed by standard reperfusion and control of extracerebral variables. Six coronal brain sections including 19 regions were examined from dogs subjected to 0 (n = 2), 5 (n = 5), 10 (n = 6), 12.5 (n = 12), 15 (n = 8), 17 (n = 5), or 20 (n = 1) minutes of cardiac arrest. Dogs were neurologically evaluated before death. RESULTS: Necrotic neurons were widespread and scattered among normal neurons. Individual regions varied in their sensitivity to different durations of cardiac arrest. There were consistent increases in the mean prevalence of necrotic neurons with increased arrest duration in the hippocampal dentate gyrus and for cerebellar granule neurons. Regionally, the caudate nucleus had the best correlation with clinical neurological deficit (rho = +.85, P < .01). CONCLUSIONS: Compared with total (summed regional) necrotic neuron prevalence scores, increased regional prevalence scores for cerebellar granule neurons with increasing arrest duration were equally significant, and scores for the caudate nucleus had nearly the same correlation with individual clinical neurological deficit. PMID- 7482663 TI - Widespread appearance of Alz-50 immunoreactive neurons in the human brain with cerebral infarction. AB - BACKGROUND AND PURPOSE: Tau-like immunoreactivity is known to develop in neurons under some experimental conditions simulating ischemia. The purpose of this study is to investigate the expression of tau-like immunoreactivity in the human brain after ischemic insult. METHODS: A series of autopsied human brains with or without ischemic lesion were investigated with immunohistochemistry (Alz-50, anti tau, and anti-ubiquitin) and with silver-staining methods (Gallyas and Bodian methods). RESULTS: Punctate immunoreactivity to Alz-50 was visualized in the cytoplasm not only of the neurons in and around the ischemic lesion but also of the neurons free from classic ischemic changes around the necrosis. Some of the neurons around the ischemic lesion were stained by the Gallyas method. Immunostaining with anti-tau and anti-ubiquitin antibodies and the conventional Bodian method failed to visualize these neurons. CONCLUSIONS: The widespread appearance of Alz-50 immunoreactive neurons during the ischemic process signifies that tau-related proteins may be related to ischemic necrosis, but the lack of neurofibrillary tangles morphologically distinguishes ischemic development of tau related proteins from the neurofibrillary degeneration in Alzheimer's disease. PMID- 7482662 TI - Neocortical neural sprouting, synaptogenesis, and behavioral recovery after neocortical infarction in rats. AB - BACKGROUND AND PURPOSE: Neuroanatomical plasticity is well described in lesions of the hippocampus but remains a subject of some controversy in the neocortex. The purpose of the present study was to measure the neocortical distribution and density of expression of proteins known to be involved in neurite growth or synaptogenesis and to correlate the neocortical expression with behavioral recovery after a focal neocortical infarction. Focal neocortical infarction creates a circumscribed lesion in the neocortex that provides a denervation stimulus for neurite growth and synaptogenesis. METHODS: Unilateral neocortical ischemia was induced in male spontaneously hypertensive Wistar rats (n = 4 per time point) by permanent occlusion of the distal middle cerebral artery and ipsilateral common carotid artery. To determine the spatial and temporal distribution of neurite growth and/or synaptogenesis, GAP-43, a growth-associated protein expressed on axonal growth cones, and synaptophysin, a calcium-binding protein found on synaptic vesicles, were examined by immunohistochemical techniques. The reaction product was measured, and the distribution was recorded. Since the resulting infarction included a portion of the forelimb neocortex, behavioral assessments of forelimb function that used the foot-fault test of Hernandez and Schallert were performed on the same rats used for immunohistochemical studies. Recovery times were 3, 7, 14, 30, and 60 days after surgery. RESULTS: Both GAP-43 and synaptophysin proteins demonstrated statistically significant increases in the density of immunoreaction product as determined by optical density measurements in the neocortex of infarcted rats compared with sham controls. The GAP-43 was elevated to statistically significant levels in forelimb, hindlimb, and parietal neocortical regions medial and lateral to the infarction only at days 3, 7, and 14. In contrast, synaptophysin demonstrated no statistically significant changes in expression at 3 or 7 days but demonstrated statistically significant increases at 14, 30, and 60 days in the forelimb, hindlimb, and parietal neocortical regions medial and lateral to the infarction as well as in the contralateral parietal neocortex. Behavioral assessment of forelimb function indicated impairment of forelimb placement on the side contralateral to the infarction that trended toward control values at 14 days and was not significantly different from controls by 30 days. CONCLUSIONS: These data support the occurrence of neurite growth followed by synaptogenesis in the neocortex, ipsilateral and contralateral to neocortical ischemia, in a pattern that corresponds both spatially and temporally with behavioral recovery. Thus, neuroanatomical remodeling in the neocortex provides a mechanism for recovery of function. PMID- 7482665 TI - Basic fibroblast growth factor increases regional cerebral blood flow and reduces infarct size after experimental ischemia in a rat model. AB - BACKGROUND AND PURPOSE: The aim of this study was to ascertain whether basic fibroblast growth factors (bFGF) caused reduction in size of cerebral infarcts in Sprague-Dawley rats with experimental ischemia. METHODS: In the first experiment we induced permanent occlusion of the left middle cerebral artery (MCA). Within 5 minutes after MCA occlusion, we infused bFGF (100 ng in 0.1 mL of saline) in the bFGF-treated group (n = 14) and 0.1 mL of saline alone in the control group (n = 7) into the common carotid artery ipsilateral to MCA occlusion. We harvested the brains 24 hours after MCA occlusion and determined infarct size planimetrically as a percentage of hemisphere size. In the second experiment cerebral blood flow (CBF) was continuously measured for 120 minutes after MCA occlusion in the bFGF treated group (n = 9) and in the control group (n = 8) with the use of laser Doppler flowmetry. RESULTS: Infarct size in the bFGF-treated group decreased significantly in comparison with that in the control group (repeated-measures ANOVA, P < .0001). CBF in the transitional areas between the MCA and the anterior cerebral artery significantly increased in the bFGF-treated group in comparison with that in the control group (repeated-measures ANOVA, P < .005). An approximate 58% decrease in infarct size and a 40% increase in regional CBF were seen on bFGF treatment. CONCLUSIONS: The present study suggested that intracarotid administration of bFGF (100 ng) can reduce infarct size after MCA occlusion. It was speculated that the increased CBF in the penumbral areas of MCA may contribute to contraction of infarct size. PMID- 7482666 TI - Time-dependent inhibition of oxotremorine-induced cerebral hyperemia by N omega nitro-L-arginine in cats. AB - BACKGROUND AND PURPOSE: Oxotremorine (OXO) is a cholinergic agonist that increases cerebral blood flow (CBF) when administered intravenously. We tested the hypothesis that OXO causes a dose-related increase in CBF in cats via a muscarinic mechanism that involves stimulation of nitric oxide synthase. METHODS: Halothane-anesthetized male cats were studied under controlled ventilation. In three groups we measured cerebral blood flow (CBF; microspheres) during 30 minutes of intravenous OXO infusion at doses of 0.5 (n = 3), 5 (n = 6), or 50 micrograms.kg-1.min-1 (n = 6). The role of muscarinic receptor activation in the CBF response to OXO (50 micrograms.kg-1.min-1) was assessed by determining the effect of atropine sulfate (2 mg.kg-1, n = 6) pretreatment in a separate group of cats. The role of nitric oxide synthase was assessed by determining the CBF response to OXO (50 micrograms.kg-1.min-1) either 30 (n = 6) or 60 minutes (n = 5) after administration of 50 mg/kg N omega-nitro-L-arginine (LNA). RESULTS: CBF to forebrain (pre-OXO, 144 +/- 12 mL.min-1.100 g-1) was unchanged with OXO 0.5 or 5 micrograms.kg-1.min-1 but increased at 10 (209 +/- 26 mL.min-1 x 100 g-1) and 30 minutes (243 +/- 35 mL.min-1 x 100 g-1) of OXO infusion at 50 micrograms.kg 1.min-1 (P < .05). Atropine sulfate prevented OXO-induced hyperemia at 10 minutes of infusion but not at 30 minutes of infusion (135 +/- 12% of pre-OXO). LNA decreased baseline CBF by approximately 50%. Treatment with LNA 30 minutes before OXO did not affect the extent of OXO-induced hyperemia (CBF, 142 +/- 15% of pre OXO at 10 minutes and 153 +/- 18% of pre-OXO at 30 minutes of OXO infusion). Treatment with LNA 60 minutes before OXO ablated OXO-induced hyperemia. CONCLUSIONS: In halothane-anesthetized cats, OXO (50 micrograms.kg-1.min-1) increases forebrain CBF by a muscarinic mechanism that involves stimulation of nitric oxide synthase. The ability of nitric oxide synthase inhibitors to block agonist-induced nitric oxide-mediated vasodilation (response to OXO) is time dependent and may not be predicted by ability of the inhibitor to significantly decrease basal CBF. PMID- 7482664 TI - Long-term high-colloid oncotic therapy for ischemic brain edema in gerbils. AB - BACKGROUND AND PURPOSE: We evaluated the effects of long-term administration of high-colloid oncotic pressure on ischemic brain edema in Mongolian gerbils. METHODS: Animals that exhibited stroke after 35 minutes of unilateral forebrain ischemia were used. The gerbils were divided into albumin- (1 g/kg body wt, 25% albumin; n = 30) and saline-injected (4 mL/kg; n = 30) groups. Both agents were administered intravenously every 12 hours starting immediately after the recirculation. Plasma colloid oncotic pressure, serum sodium and potassium concentrations, and brain water, sodium, and potassium content were measured 24, 48, and 72 hours after recirculation. RESULTS: Plasma colloid oncotic pressure at 24, 48, and 72 hours after recirculation was significantly higher in the albumin- (26.1 +/- 2.3 mm Hg) than in the saline-treated group (18.5 +/- 1.9 mm Hg; P < .01), and brain water content of the ischemic hemisphere was significantly lower in the albumin group (79.5%, 80.2%, and 80.5%, respectively) than in the saline group (80.9%, 81.6%, and 82.1%, respectively; P < .05) at all three time points. Brain sodium content at 24 hours was significantly lower in the albumin than in the saline group (P < .05), while brain potassium content at 24 and 48 hours was significantly higher in the albumin than in the saline group (P < .05). The changes in brain water and sodium plus potassium content, which were calculated from differences between the ischemic and nonischemic hemispheres, showed a significant correlation in both groups (P < .01), but there was no significant difference between the linear regression lines for both groups. CONCLUSIONS: Long term high-colloid oncotic pressure was effective in treating ischemic brain edema, probably acting by diminishing the bulk flow through the disrupted blood brain barrier and ameliorating the vasogenic edema. PMID- 7482667 TI - Hemodilution accelerates the passage of plasma (not red cells) through cerebral microvessels in rats. AB - BACKGROUND AND PURPOSE: Hemodilution lowers the total circulatory red cell mass and blood viscosity and thereby may alter the time of passage of red cells and plasma through cerebral microvessels. This study was designed to clarify this question. METHODS: Adult Wistar-Kyoto rats, aged approximately 32 weeks, were divided into hemodilution and control groups. Local cerebral blood flow and microvascular red cell and plasma volumes in 14 brain structures were measured with the use of [14C]iodoantipyrine, 55Fe-labeled red cells, and [14C]inulin, respectively. RESULTS: In the control group, the hematocrit in cerebral microvessels ranged from 0.29 to 0.45 with a mean of 0.36, which was 71% of the systemic hematocrit (0.51). The mean transit times of blood, red cells, and plasma through microvessels were 0.62 to 1.77 seconds (mean, 0.92 second), 0.44 to 1.15 seconds (mean, 0.65 second), and 0.78 to 2.5 seconds (mean, 1.25 seconds), respectively. In the hemodilution group, the mean hematocrit in microvessels was 0.28, which was 89% of the systemic hematocrit (0.32). Local cerebral blood flow was approximately 59% higher (P < .01) than that of the control animals. The rate of oxygen delivered to the brain was slightly increased (9%) after hemodilution. Blood volume in cerebral microvessels was similar to that of the control group. Mean transit time of blood was 0.62 second (68% of the control), transit time of red cells was 0.53 second (85% of the control), and transit time of plasma was 0.67 second (54% of the control). CONCLUSIONS: These findings indicate that isovolemic hemodilution accelerates the plasma (not red cell) flow velocity in cerebral microvessels. PMID- 7482669 TI - Failure of isradipine to reduce infarct size in mouse, gerbil, and rat models of cerebral ischemia. AB - BACKGROUND AND PURPOSE: The dihydropyridine L-type calcium channel blocker isradipine has been reported to exhibit neuroprotective properties in some, but not all, studies performed in the rat middle cerebral artery occlusion (MCAO) model. In the present study, we examined isradipine in several other models of focal and global ischemia: rat rose bengal, mouse MCAO, and gerbil bilateral carotid artery occlusion (BCAO). For comparison, a novel calcium channel blocker, SB201823A, that we have previously shown to be neuroprotective in rat and gerbil models was also examined in the mouse. METHODS: In the gerbil BCAO model, isradipine was administered at 2.5 mg/kg i.p. as a single dose 60 minutes after ischemia (n = 10). Corresponding controls received vehicle (n = 10), and sham operated animals received no treatment (n = 6). Locomotor activity and histological assessments were made at 4 days after ischemia. In the rat photothrombotic occlusion model, isradipine was administered at 2.5 mg/kg i.p. as a single dose 60 minutes after ischemia (n = 10), and corresponding controls (n = 10) received vehicle. Histological assessment was made at 7 days after ischemia. In the mouse MCAO model, isradipine was also administered at 2.5 mg/kg i.p. as a single dose 60 minutes after ischemia. Histological assessments were made at 1 (n = 13), 2 (n = 9), and 4 (n = 9) days after ischemia. Vehicle numbers were n = 10, n = 6, and n = 8, respectively. Isradipine and SB201823A were also examined using a combined preischemia and postischemia regimen. Isradipine was administered at 2.5 mg/kg i.p. before occlusion, 1.25 mg/kg i.p. 1 hour after occlusion, 1.25 mg/kg i.p. 2 hours after occlusion, and 2.5 mg/kg twice a day for 3 days after occlusion (n = 16). Corresponding controls received vehicle at the same time points (n = 14). SB201823A was administered 30 minutes before occlusion, 30 minutes after occlusion, and twice daily for 3 days (n = 12). Corresponding controls received vehicle (n = 9). Histological assessment was performed at 4 days after ischemia. RESULTS: When given after ischemia, isradipine failed to affect lesion volume in both the rat and mouse models. In the gerbil, locomotor hyperactivity and hippocampal cell loss were unaffected. Given before and after ischemia in the mouse, isradipine was also ineffective, whereas SB201823A produced a significant reduction in lesion volume. CONCLUSIONS: The L-type calcium channel blocker isradipine was devoid of neuroprotective activity in focal and global models of cerebral ischemia in three species of normotensive animals. These results were compared with data for the novel calcium channel blocker SB201823A, which exhibited a significant effect after pre- and postocclusion administration in the mouse model of permanent focal ischemia. PMID- 7482672 TI - Cystatin C mutation in an elderly man with sporadic amyloid angiopathy and intracerebral hemorrhage. AB - BACKGROUND: Cerebral amyloid angiopathy (CAA) with intracerebral hemorrhage (ICH) occurs both sporadically and as a result of mutations in either cystatin C or the amyloid precursor protein. ICH due to cystatin C mutations typically occurs in young people of Icelandic origin. CASE DESCRIPTION: We report a case of sporadic CAA with ICH in an elderly Croatian man with a mutation in cystatin C identical to that found in Icelandic hereditary cerebral hemorrhage with amyloidosis. CONCLUSIONS: This is the first case report of sporadic CAA associated with the same mutation causing hereditary cerebral hemorrhage with amyloidosis of the Icelandic type. Sporadic CAA may thus be associated with genetic mutations in some patients. The frequency of these mutations is yet to be determined. PMID- 7482670 TI - Cerebral microembolism and the risk of ischemia in asymptomatic high-grade internal carotid artery stenosis. AB - BACKGROUND AND PURPOSE: Previous work has shown that cerebral microembolism detected with transcranial Doppler sonography distal to internal carotid artery stenosis occurs more frequently in recently symptomatic compared with asymptomatic patients. It has remained unclear whether cerebral microembolism also indicates a higher risk of future cerebral or retinal ischemia. SUMMARY OF REPORT: Sixty-four asymptomatic patients with unilateral 70% to 90% internal carotid artery stenosis were investigated prospectively (mean follow-up, 72 weeks). Five patients developed ischemic symptoms attributable to the stenosis (transient ischemic attack, 2 patients; stroke, 3 patients). A microembolic rate of > or = 2 per hour in the ipsilateral middle cerebral artery was associated with a substantially increased risk of developing ischemia of the corresponding carotid territory (odds ratio, 31; 95% confidence interval, 3 to 302; P = .005). CONCLUSIONS: This prospective pilot study suggests that cerebral microembolism detected with transcranial Doppler sonography may define a high-risk subgroup among patients with asymptomatic high-grade internal carotid artery stenosis. PMID- 7482671 TI - Massive persistent release of excitatory amino acids following human occlusive stroke. AB - BACKGROUND: Animal stroke models demonstrate excitatory amino acid (EAA) release in ischemic tissue, as measured by microdialysis. Currently glutamate antagonist drugs are being developed to protect brain tissue after ischemic events. However, the role of EAAs in human occlusive stroke is not well known. We therefore measured glutamate and aspartate release in a patient after occlusive stroke. CASE DESCRIPTION: We describe a case of occlusive stroke in a 50-year-old man. A partial temporal lobectomy was done to remove infarcted tissue and to prevent brain stem compression as well as uncal herniation. A microdialysis probe was placed into the cortex to measure EAAs. Massively increased levels of glutamate and aspartate were detected in the extracellular fluid in this patient (> 300 times normal levels 6 days after infarction). CONCLUSIONS: These findings indicate that EAAs are tremendously increased in brain tissue after occlusive stroke. The time course of the release of EAAs is much longer than animal studies have suggested previously. Administration of EAA antagonists to patients with ischemic stroke may therefore be beneficial. PMID- 7482673 TI - Delayed onset of fatal basilar thrombotic embolus after whiplash injury. AB - BACKGROUND: Whiplash injuries are generally seen after rear-end rather than frontal car collisions. Previous reports have documented death up to 8 days after serious whiplash injury. We report a case of lethal basilar thrombotic embolus that occurred 2 months after the patient's injury in a collision. CASE DESCRIPTION: After whiplash trauma in a car accident, a 50-year-old taxi driver suffered from headache and episodic visual disturbances. Two months after the accident he suddenly lost consciousness and was admitted to the hospital. A CT scan performed at that time was indicative of basilar thrombosis. The patient died 3 days later. The autopsy revealed a thrombosis in the right vertebral artery and a thrombotic embolus in the basilar artery. Microscopically, a lesion of the right vertebral artery was found at the level of the atlantoaxial joint. CONCLUSIONS: We conclude that the whiplash injury caused a lesion of the right vertebral artery, leading to repeated transient ischemic attacks and finally to a fatal basilar thrombotic embolus. We suggest that in patients with disturbances of the vertebrobasilar circulation, attention should be paid to occurrence of neck trauma in the preceding 3 months. Further, anticoagulant therapy should particularly be considered in patients who after suffering neck injuries develop signs of transient ischemic attacks with origin from the posterior cerebral circulation. PMID- 7482674 TI - Measurement of instrumental activities of daily living in stroke. PMID- 7482668 TI - 7-Nitroindazole inhibits brain nitric oxide synthase and cerebral vasodilatation in response to N-methyl-D-aspartate. AB - BACKGROUND AND PURPOSE: N-Methyl-D-aspartate (NMDA) produces dilatation of cerebral arterioles that is dependent on production of nitric oxide (NO). In these experiments we examined the hypothesis that cerebral vasodilatation in response to NMDA is mediated by the neuronal isoform of NO synthase. METHODS: We measured diameters of cerebral arterioles (baseline diameter, 89 +/- 7 microns) using a closed cranial window in anesthetized rabbits that received either vehicle (10 mL/kg IP peanut oil) or 7-nitroindazole (7-NI; 50 mg/kg IP). 7-NI is reported to be a selective inhibitor of neuronal NO synthase. RESULTS: Two hours after administration of 7-NI, activity of brain NO synthase (measured by conversion of L-arginine to L-citrulline) was reduced by 33% compared with vehicle (24 +/- 1 versus 16 +/- 3 pmol/min per milligram protein; n = 7; P < .05). Dilatation of cerebral arterioles in response to NMDA (100 and 300 mumol/L) was inhibited by 30% to 40% by 7-NI compared with responses in the presence of vehicle (23 +/- 6% versus 14 +/- 5% and 30 +/- 4% versus 21 +/- 5%, respectively; P < .05 for both concentrations; n = 10). In contrast, vasodilatation in response to acetylcholine (1 mumol/L) was similar in vehicle- and 7-NI-treated animals (17 +/- 5% versus 21 +/- 4%; P > .05). CONCLUSIONS: These findings suggest that vasodilatation in response to NMDA is mediated by neuronally derived NO. 7-NI appears to produce selective inhibition of brain NO synthase but not endothelial NO synthase. PMID- 7482675 TI - Neurobiology of autonomic and thermal sensory symptoms and dysfunction after stroke. PMID- 7482676 TI - Recurrent cerebellar hematomas. PMID- 7482677 TI - Women's education and fertility: results from 26 Demographic and Health Surveys. AB - This article presents an updated overview of the relationship between women's education and fertility. Data from the Demographic and Health Surveys for 26 countries are examined. The analysis confirms that higher education is consistently associated with lower fertility. However, a considerable diversity exists in the magnitude of the gap between upper and lower educational strata and in the strength of the association. In some of the least-developed countries, education might have a positive impact on fertility at the lower end of the educational range. Yet, compared with patterns documented a decade ago, the fertility-enhancing impact of schooling has become increasingly rare. The study also examines the impact of female education on age at marriage, family-size preference, and contraceptive use. It confirms that education enhances women's ability to make reproductive choices. PMID- 7482678 TI - Self-reported symptoms of gynecological morbidity and their treatment in south India. AB - This article presents an analysis of self-reported symptoms of gynecological problems among 3,600 recent mothers in Karnataka State, India. Approximately one third of all women reported at least one current symptom; the most common were a feeling of weakness and tiredness (suggestive of anemia); menstrual disorders; white or colored vaginal discharge (suggestive of lower reproductive tract infection); and lower abdominal pain and discharge with fever (suggestive of acute pelvic inflammatory disease). Obstetric morbidity, associated with the last live birth, was strongly predictive of current gynecological symptoms. Women who delivered their last child in a private institution were significantly less likely to report symptoms than were those who delivered at home or in a government hospital. Nonusers or users of reversible contraceptive methods were also less likely to report symptoms of morbid conditions than were sterilized women. These associations persisted in analyses controlling for potentially confounding economic and demographic characteristics, and have far-reaching policy implications. PMID- 7482681 TI - The question of access. PMID- 7482679 TI - Maternal mortality in India: estimates from a regression model. AB - This report outlines a new technique for the estimation of maternal mortality by relating the sex differentials in mortality for people of reproductive age to the age schedule of fertility. The application of this method to the data from the Sample Registration System for 1982-86 indicates a level of maternal mortality of 580 deaths per 100,000 live births for India as a whole, 638 deaths in rural areas, and 389 deaths in urban areas. Estimates derived for the major states suggest relatively high maternal mortality in the eastern and northern parts of the country. They also indicate a substantial decline in maternal mortality since the 1960s. The decline in the birth rate is estimated to have accounted for nearly one-fourth of the decrease in the maternal death rate and 5 percent of the fall in the maternal mortality ratio in the 10-year period between 1972-76 and 1982-86. The method of estimation described here is well-suited to the data circumstances in India. PMID- 7482680 TI - The effect of the sex of interviewers on the quality of data in a Nigerian family planning questionnaire. AB - Traditionally, female interviewers have been preferred to men for conducting fertility and family planning surveys. However, in West Africa, evidence for their superiority over male interviewers is mixed. In Nigeria, as part of a four state pretest of the national family planning questionnaire, an experimental design was incorporated to quantify effects of the sex of the interviewer. In one state, reinterviews were also performed to measure the reliability of responses. In the conservative northern state of Kano, the use of male interviewers was problematic. However, in the other three states, only weak evidence was found to mitigate against the use of male interviewers. In fact, in two states, the proportion of respondents reporting knowledge of several contraceptive methods was significantly higher when male interviewers conducted the survey. Respondents' reports of contraceptive use were very unreliable for interviewers of both sexes. PMID- 7482682 TI - Ghana 1993: results from the Demographic and Health Survey. PMID- 7482683 TI - Vancomycin pharmacokinetics and dosing in premature neonates. AB - We prospectively studied the pharmacokinetic parameters of vancomycin in premature neonates given vancomycin according to a dosage protocol developed in our neonatal unit. Study infants were administered vancomycin according to four postconceptional age (PCA) groups: (0) 18 mg/kg every 36 h for PCA < 27 weeks; (I) 16 mg/kg every 24 h for PCA 27-30 weeks; (II) 18 mg/kg every 18 h for PCA 31 36 weeks; and (III) 15 mg/kg every 12 h for PCA > or = 37 weeks. Pharmacokinetic parameters were calculated from peak and trough serum vancomycin concentrations at steady state. Results in 44 infants (PCA, 27-44 weeks) showed that our dosage regimen achieved target peak serum vancomycin concentrations in 64% of neonates in Groups I-III, although it tended to undershoot the target trough concentrations. Volume of distribution (Vd), normalized for body weight, remained constant throughout the PCA range, with a mean value of 0.56 L/kg, whereas absolute clearance (r = 0.81) and normalized clearance (r = 0.48) increased with PCA (p < 0.005). The increase in clearance with PCA is associated with a greater elimination rate constant and shorter half-life. Vancomycin therapy can be initiated in a standard fashion according to our protocol or by individualizing the dosage regimen based on a Vd of 0.56 L/kg and clearance estimated from the infant's body weight and PCA groups. PMID- 7482684 TI - Prediction of free levels of phenytoin and carbamazepine in patients comedicated with valproic acid. AB - Serum samples from patients receiving phenytoin (PHT) or carbamazepine (CBZ) and sodium volproate (VPA) polytherapy were filtered at 37 degrees C and the free concentrations were measured by high-performance liquid chromatography. The mean apparent dissociation constants (KD) for the binding of PHT and CBZ to albumin were calculated. The mean KD values were used to predict free levels of PHT or CBZ in serum from patients also taking VPA, based on information about the total serum concentrations of the drugs and albumin. In the presence of VPA the values of the free fraction (alpha) for PHT and CBZ increased and the values for the apparent dissociation constants (KD) decreased compared with the situation of monotherapy. These decreases in the value of KD and increases in the value of alpha were statistically significant. The correlations between values for predicted and analysed free levels of PHT or CBZ were close, with r values of 0.999 and 0.998, respectively. The mean prediction error was small, 0.01 +/- 0.22 mumol/L for PHT and 0.03 +/- 0.42 mumol/L for CBZ. The percentage differences between predicted and observed free levels for PHT and CBZ were < 10%. We conclude that our method of prediction is sufficiently accurate and precise for clinical use. PMID- 7482685 TI - Inhibition of trazodone metabolism by thioridazine in humans. AB - To clarify the involvement of cytochrome P4502D6 (CYP2D6) in the metabolism of trazodone, the effects of coadministration of thioridazine, which is an inhibitor of this isozyme, on plasma concentrations of trazodone and its active metabolite m-chlorophenylpiperazine (m-CPP) were studied. The subjects were 11 depressed patients receiving trazodone at bedtime for 1-18 weeks. The dose was 150 mg in 10 patients and 300 mg in one. Thioridazine 40 mg/day was coadministered for 1 week, and blood samplings were performed before and after the coadministration. Thioridazine significantly (p < 0.001) increased plasma concentrations of both trazodone (713 +/- 252 vs. 969 +/- 370 ng/ml) and m-CPP (61 +/- 22 vs. 94 +/- 34 ng/ml). The present study thus suggests that CYP2D6 is involved in the metabolism of trazodone. PMID- 7482686 TI - Pharmacokinetics of propofol infusion in Asian patients undergoing coronary artery bypass grafting. AB - The pharmacokinetics of propofol was studied in 11 Asian patients with fentanyl isoflurane anaesthesia during cardiopulmonary bypass (CPB) and undergoing elective coronary artery bypass grafting (CABG). Instead of the usual increments of morphine and a benzodiazepine, propofol (4 mg/kg/h) was initiated at the start of CPB and ceased at CPB separation. Whole blood propofol concentrations were determined during and postinfusion using high-performance liquid chromatography with fluorescence detection. Data from four patients seemed to fit a two compartment model, whereas those from seven patients were significantly (F test, p < 0.05) better fitted to a three-compartment model. The pharmacokinetic parameters were as follows: The mean (SD) of the initial distribution phase t1/2 pi, intermediate distribution phase t1/2 alpha, and elimination phase t1/2 beta were 2.22 (1.04) min, 42.9 (16.4) min, and 370 (138) min, respectively. The mean clearance of 1.31 (0.50) L/min was lower than those reported from other studies, whereas the mean blood concentration of 2.2 (1.0) mg/L at the 1-h infusion period was higher. The mean calculated apparent Css was 3.9 (1.5) mg/L. The low clearance is likely to be due to hemodynamic changes during CPB and CABG, thereby affecting drug distribution and blood flow to the liver. PMID- 7482687 TI - Quality assurance program for cyclosporin G (OG37-325). AB - Cyclosporine-G (CsG) (OG37-325) an analogue of cyclosporine-A (CsA) is presently undergoing clinical trials. As therapeutic monitoring of CsG was an integral part of these evaluations, the performance of laboratories measuring the drug was assessed through an external quality assurance program, which included all North American centers participating in the trials. The assays used by participating centers were high-performance liquid chromatography (HPLC) (n = 3), radioimmunoassay (RIA) (n = 5), and fluorescence polarization immunoassay (FPIA) (n = 24). The latter two assays, developed for measurement of CsA, were adapted for measurement of CsG. The FPIA was the most precise, with the largest proportion of laboratories reporting coefficients of variation (CNs) of < 10%. RIA was the least accurate method, with approximately 74% of results differing by > 20% from target values. FPIA and RIA methods exhibited mean recoveries of CsG of 112 and 129%, respectively. The ranking of the specificity of the assays from measurement of parent drug was HPLC > RIA > FPIA. FPIA and RIA produced values that were, on average, approximately 60% higher than those reported by HPLC in pooled whole blood specimens obtained from renal transplant patients. It is recommended that if CsG is approved for routine clinical use, then assays developed specifically for measurement of the drug be used. PMID- 7482689 TI - High-performance liquid chromatographic evaluation of the effect of heat treatment on trimethoprim and sulfamethoxazole stability in serum. AB - Heat treatment of patient samples is utilized as a method to decrease the risk of accidental transmission of human immunodeficiency virus (HIV). Heat treatment has been reported to affect the outcome of drug analysis. In this study, the effects of heat treatment (56 degrees C for 5 h) and storage for 2 months at -20 degrees C on the stability of trimethoprim (TMP) and sulfamethoxazole (SMX) at three different concentrations in serum (10/100, 1/20, and 0.1/5 microgram/mL) each were evaluated. Simultaneous determination of TMP, SMX, and sulfamethazine (SMeth), the internal standard, in serum was performed by a reversed-phase high performance liquid chromatographic (HPLC) procedure with isocratic elution and ultraviolet detection. The peak/height ratios (PHRs) for each sample from untreated and heat-treated groups were compared. No statistically significant differences were found between untreated and heat-treated groups for TMP. Heat treatment decreased the PHR for SMX at 100 micrograms/ml concentration (p = 0.042) and increased the PHR for SMX at 20 micrograms/ml concentration (p = 0.049). These marginal differences are unlikely to be significant. Storage of samples for 2 months at -20 degrees C had no statistically significant effect on sample PHRs. Thus, heat treatment of serum does not alter clinical interpretation of TMP and SMX at clinically relevant concentrations and may protect laboratory workers from accidental HIV exposure. PMID- 7482690 TI - Stability of tacrolimus (FK 506) and cyclosporin G in whole blood. AB - The stability of two new immunosuppressants, tacrolimus and cyclosporin G (CsG), was evaluated in whole blood following incubation of patient specimens at ambient temperature or 4 degrees C for time periods of 2 (48 h) and 7 days (168 h) after collection. No decrease in CsG concentrations was noted over a 7-day period for specimens stored at either ambient temperature or 4 degrees C. For tacrolimus, the concentrations of drug in whole blood stored at ambient temperature for 7 days had a slight downward trend. At 7 days, the ratio of tacrolimus concentrations to day zero concentrations ranged from 0.86 to 1.15, with a mean decrease of 5%. However, these changes are within the published precision characteristics of the tacrolimus immunoassay. Evaluation of a smaller group of whole blood specimens provided evidence of adequate stability of these two drugs over a 13-day period. PMID- 7482688 TI - Validation of a quick modeling program generating clearance estimates at steady state for routine therapeutic drug monitoring. AB - Therapeutic drug monitoring (TDM) of chronic treatments is justified for several reasons, including relative over- or underdosage due to variable individual elimination, pharmacokinetic interactions in drug combinations, and noncompliance. In all these circumstances, the prescribing physician is interested in having an estimation of the patient's clearance of the drug, even from one measurement. We compare a validated bayesian program, USC*Pack of Jelliffe, found difficult to use in daily routine, with a "home-made" program. The latter, which is capable of taking data from a clinical database, will generate a graphic simulation of daily plasma drug concentrations together with an estimation of steady-state clearance more rapidly than does USC*Pack. Both programs were run with only one measured plasma level. The patients were 83 children or young adults treated with phenobarbital (PB), carbamazepine (CBZ), and/or Valproic acid (VPA) who were resistant to monotherapy and who were to be sampled two to four times between doses. Drugs were routinely assayed by high performance liquid chromatography (HPLC). Despite the rough character of Phacile (numeric integration and adjustment of only two of three parameters, without an acknowledged minimization algorithm), the results are comparable to those obtained with USC*Pack for estimating clearance and predicting plasma drug concentrations. Phacile algorithm, although simple, has proven of interest in routine TDM and as an introduction for medical students to the bayesian approach of population pharmacokinetics. PMID- 7482691 TI - An improved micromethod for vancomycin determination by high-performance liquid chromatography. AB - A micromethod using reversed-phase high-performance liquid chromatography for the analysis of vancomycin in human serum or plasma was developed. Ristocetin was used as the internal standard. Chromatographic conditions included an amino propyl column, a mobile phase with 62% acetonitrile and 38% sodium phosphate buffer (pH 7.0), a total run time of 10 min, and ultraviolet absorbance detection at 225 nm. Multilevel calibration was found to be linear between 1.0 and 100 micrograms/ml with correlation coefficients of the calibration line slope consistently > 0.999. Recovery of vancomycin from serum was nearly complete, and no interference from commonly used drugs was observed. This procedure is simple, sensitive, rapid, precise, selective, and requires only 50 microliters of serum or plasma for completion. PMID- 7482692 TI - Quantitative determination of E- and Z-doxepin and E- and Z-desmethyldoxepin by high-performance liquid chromatography. AB - A high-performance liquid chromatographic (HPLC) method was developed for the quantitative, simultaneous determination of the following four compounds in serum: E-doxepin, Z-doxepin, E-desmethyldoxepin, and Z-desmethyldoxepin. A 3 microns analytical silica column (6 x 100 mm) was employed with the mobile phase 0.025 M phosphate:acetonitrile:n-nonylamine (80:20:1). This HPLC method allows for the accurate measurement of all four isomeric compounds. PMID- 7482693 TI - Binding of 17-alpha-methyltestosterone in vitro to human sex hormone binding globulin and rat ventral prostate androgen receptors. AB - We measured the relative binding affinity (RBA) of 17-alpha-methyltestosterone (MET) for the rat prostate androgen receptor and human sex hormone-binding globulin (SHBG) in vitro. The reference compound in both instances was dihydrotestosterone (DHT), the RBA of which was set at 100%. The RBA of methyltestosterone was 44% for the androgen receptor, but 11% for SHBG. The androgen receptor, therefore, would appear to recognize a different part of an androgen compared to the recognition site on SHBG. The difference in RBAs may amplify the biologic action of methyltestosterone. PMID- 7482694 TI - Is routine measurement of free phenytoin concentrations rational? PMID- 7482695 TI - Modular design of components of the operational RNA code for alanine in evolution. PMID- 7482696 TI - No soup for starters? Autotrophy and the origins of metabolism. AB - In recent years, several alternatives to the classical prebiotic-soup model for the origin of life have been proposed. Among these, the theory of Wachtershauser proposes that an archaic version of the autotrophic reductive citric acid cycle, driven by pyrite formation and contained on the resulting pyrite surface, was the earliest metabolic cycle, from which the central biosynthetic pathways arose, initially without enzymes or nucleic acids. PMID- 7482697 TI - The death domain: a module shared by proteins with diverse cellular functions. PMID- 7482698 TI - DNA polymerase beta belongs to an ancient nucleotidyltransferase superfamily. PMID- 7482699 TI - The FHA domain: a putative nuclear signalling domain found in protein kinases and transcription factors. PMID- 7482700 TI - Circular permutations of protein sequence: not so rare? PMID- 7482701 TI - Origin of PDZ (DHR, GLGF) domains. PMID- 7482702 TI - Protein splicing: self-splicing of genetically mobile elements at the protein level. AB - Protein splicing is a newly discovered process that is the protein equivalent of RNA splicing. Protein splicing proceeds through a branched protein intermediate, and in vitro studies indicate that the reaction is autocatalytic. The excised 'intein' proteins are site-specific DNA endonucleases that catalyse genetic mobility of their DNA coding sequence by an 'intein homing' mechanism. PMID- 7482703 TI - Biochemical and genetic dissection of mitotic chromosome condensation. AB - The molecular mechanisms responsible for mitotic chromosome condensation are unknown. Two independent approaches, biochemical studies in vertebrate cells and genetic analyses in yeasts, have converged recently, leading to the identification of a family of putative ATPases that play a fundamental role in this process. Further characterization of these proteins promises to uncover a highly dynamic aspect of mitotic chromosome architecture. PMID- 7482704 TI - Cell signalling by integrins and immunoglobulin receptors in primed neutrophils. AB - Neutrophils use a variety of cell-surface receptors for attachment to surfaces, such as the endothelium or opsonized pathogens. During inflammation or infection, chemoattractants or cytokines bind to neutrophils and upregulate or 'prime' their responsiveness. Priming can increase the number of some receptors expressed on the cell surface but can also change receptor function, so that the receptor has altered ligand-binding properties or becomes linked to new intracellular signalling systems. PMID- 7482705 TI - GPI-anchor biosynthesis. AB - Many eukaryotic proteins bind to membranes using a glycosylphosphatidylinositol (GPI) anchor. GPI anchors are essential in yeasts and probably also in protozoa. Although mammalian cells can survive without GPI anchors, their deficiency in haemopoietic cells cause a haemolytic disease, paroxysmal nocturnal haemoglobinuria (PNH). Here, we discuss recent progress in our understanding of GPI-anchor biosynthesis that could lead to a better understanding of PNH and chemotherapeutic agents to treat protozoal and fungal infections. PMID- 7482707 TI - RASMOL: biomolecular graphics for all. PMID- 7482708 TI - How the molten globule became. PMID- 7482706 TI - Vectorette, splinkerette and boomerang DNA amplification. AB - Methods and reagents is a unique monthly column that highlights current discussions in the newsgroup bionet.molbio.methods-reagents, available on the Internet. This month's column discusses some methods of amplifying DNA other than by using the standard polymerase chain reaction (PCR). For details on how to partake in the newsgroup, see the accompanying box. PMID- 7482709 TI - Pharmacologically induced regression of chronic transplant rejection. AB - Chronic rejection, characterized by a progressive obliterative arteriopathy, is a major cause of graft failure in long-surviving human transplants for which there is no effective treatment. Leflunomide, an isoxazol derivative, has been shown to be a novel immunomodulatory drug that profoundly suppresses the immune response. In this study, 58 Fisher-344 rats received cardiac transplantation from Lewis rats. All the recipients were given CsA at 2.5 mg/kg for 5 days postoperatively. Without further treatments, the arterial intima was progressively injured by mononuclear cell infiltration and Ab deposition. Smooth muscle cell and fibroblast proliferation in the intima became a predominant phenomenon by day 90. CsA was ineffective in controlling the progress of arterial intimal thickening when treatment began on day 30. Leflunomide at 5 mg/kg failed to control arterial intimal thickening by day 60 when therapy began on day 30. However, the progress of arterial intimal thickening was significantly inhibited by day 90 when the dosage of leflunomide had been increased to 10 mg/kg on day 60. Combined therapy with leflunomide and CsA at 5 mg/kg for 30 days dramatically reversed the arterial thickening by day 60. After increasing the dosages of both leflunomide and CsA to 10 mg/kg on day 60, the combination therapy steadily controlled the chronic rejection. Only the combination therapy significantly down-regulated circulating antidonor IgM and IgG titers. In rat smooth muscle cell culture, this same drug combination had a synergistic inhibitory effect on proliferation. Therefore, the combination therapy of leflunomide and CsA could reverse and control the progress of chronic rejection, while leflunomide, at higher dosage as a monotherapy, could stabilize chronic rejection in this model. The mechanism of the regression of chronic rejection by leflunomide and cyclosporine may be related to their in vitro abilities to control not only lymphocyte but smooth muscle cell proliferation, as well. The synergistic effect of these two drugs on vascular smooth muscle cell proliferation in vitro may be an important part of this novel activity. This unique feature holds intriguing possibilities for treating established chronic rejection. PMID- 7482710 TI - Evidence that graft coronary arteriosclerosis begins in the early phase after transplantation and progresses without chronic immunoreaction. Histopathological analysis using a retransplantation model. AB - The histopathological features of chronic rejection and its initiation were assessed using rat heterotopic heart transplantation and retransplantation models. Fully allogeneic or minor, non-MHC antigen-mismatch heart grafts transplanted into recipient rats treated with a short course of FK506 showed long term survival but developed graft atherosclerosis after 40 days posttransplantation. Retransplantation of allografts back into the original donor strain did not prevent graft atherosclerosis if the grafts had resided in the primary recipient for up to 5 days; residence in the primary allogeneic recipient for less than 4 days did not result in graft atherosclerosis in the secondary recipient. Short-course administration of FK506 did not affect the production of these changes. Graft coronary arteriosclerosis begins between 3 and 5 days posttransplantation and progresses without continuous allogeneic immunological drive. The present findings will provide a new means by which to approach the analysis of development of chronic allograft rejection. PMID- 7482711 TI - Ischemic preconditioning improves cardiac functional recovery following preservation with University of Wisconsin solution. AB - Single or multiple brief periods of regional or global ischemia and reperfusion prior to a prolonged ischemic insult showed cardioprotective effects. Although this phenomenon (ischemic preconditioning [IPC]) has been described in ischemic reperfusion models, the effect of IPC on heart preservation has not been previously reported. We, thus, investigated the effect of IPC on heart preservation. Hearts isolated from male Wistar rats (250-350 g) were mounted on a Langendorff apparatus to estimate baseline function (aortic flow, coronary flow, cardiac output, heart rate, systolic pressure, and rate pressure product). All hearts were divided into 5 groups. In groups 1 and 4, the hearts were subjected to 8 and 12 hr of preservation, respectively. The hearts in group 2 were subjected to a single 2.5-minute cycle of normothermic global ischemic episode (IPC) before 8 hr of preservation. In groups 3 and 5, the hearts were subjected to two 2.5-min IPC cycles and stored for 8 or 12 hr. The hearts were arrested with University of Wisconsin solution and stored at 4 degrees C. Following storage, the hearts were reperfused and measured postpreservative function to assess cardiac functional recovery. Lactate and troponin-T leakage in the coronary perfusate was also measured. In group 3, the treatment of two 2.5-min IPC cycles significantly increased cardiac output, but the treatment of single 2.5-min IPC cycle did not affect the result. In the extended preservation group (group 5), the recovery (%) of both coronary flow and cardiac output were significantly increased compared with group 4. Furthermore, lactate leakage was significantly reduced in groups 2 and 3. These results suggest that IPC improves cardiac functional recovery following simple cold storage and has cardioprotective effects in rat heart preservation. PMID- 7482712 TI - Reproducibility of the Banff classification of renal allograft pathology. Inter- and intraobserver variation. AB - The present study was undertaken to investigate the inter- and intraobserver variation in use of the scoring system for glomerulitis, vasculitis, interstitial inflammation, tubulitis and arteriolar hyalinosis that is an essential part of the recently proposed Banff classification of renal allograft biopsies. Seventy seven biopsies done less than 90 days after transplantation were included. The scoring was done blindly by five pathologists on biopsies stained with H&E and PAS. The volume fraction of interstitial inflammation was estimated. Spearman rank correlation coefficient and kappa values were used for the evaluation of reproducibility. The results of both inter- and intraobserver variability showed a good correlation and reasonable kappa values for vasculitis, interstitial inflammatory infiltration, and tubulitis. Less-good correlation was found for glomerulitis and arteriolar hyalinosis. The interobserver kappa score for grading of the rejection severity was 0.40 overall but 0.56 when only presence or absence of acute rejection was considered and 0.66 for presence or absence of vasculitis. Weighted kappa values for interobserver vasculitis score and rejection grading were 0.58 and 0.55, respectively. A strong association existed between the volume fraction of interstitial inflammation and the semiquantitative scoring for interstitial inflammation. In conclusion, the good correlations for the key elements in the grading of the allograft biopsies in the present classification system, confirmed the utility of the defined criteria for grading rejection. More precisely defined criteria or simplification of the scoring system are needed for glomerulitis and arteriolar hyalinosis--parameters not used in the diagnosis of rejection. PMID- 7482713 TI - The impact of immunosuppressive regimens on the cost of liver transplantation- results from the U.S. FK506 multicenter trial. AB - In an effort to determine the total one-year cost of liver transplantation, the underlying drivers of that cost, and any cost differences between alternative immunosuppressive regimens, an analysis was performed comparing the average one year posttransplant charges of 322 patients participating in the "U.S. Multi center Prospective Randomized Trial Comparing FK-506 to Cyclosporine in Liver Transplantation." Total one-year inpatient charges including all readmissions were examined. Professional fees and outpatient charges were excluded. Costs for tacrolimus drug and blood assays were assumed to be equal to those in the CsA group. For patients completing the study, the tacrolimus group had an average length of stay and average one-year cost seven days (P = .06) and $19,290 (P = .05) lower than the CsA group. The difference in rejection profiles between the two arms seems to largely account for the lower costs. The tacrolimus arm consistently had fewer patients in the more severe rejection groups. Increased incidence and severity of rejection were directly related to higher average lengths of stay and costs of transplantation (P < .001). Tacrolimus immunosuppression during the first year after liver transplantation is more cost effective than CsA in achieving similar patient and graft survival rates. Differing incidence and severity of rejection can dramatically affect the first year cost of liver transplantation. PMID- 7482714 TI - Orthotopic liver transplantation for Crigler-Najjar type I disease in six children. AB - Hyperbilirubinemia in Crigler-Najjar disease type I (CN) can be partially controlled by daily phototherapy, but these children remain at permanent risk of developing brain damage due to kernicterus. Because liver transplantation is the only available curative treatment for liver-based inborn errors of metabolism, orthotopic liver transplantation (OLT) was performed in six patients with CN. Mean age at surgery was 52.5 months (range 27 to 100). Despite a mean daily phototherapy of 12.4 +/- 0.8 hr, mean bilirubin of the 6 patients was 388 microM/L (range 175 to 703) before OLT; one of them was also being treated with tin-protoporphyrin. All 6 had elevated AST/ALT, ranging from 1.4 to 6 times upper normal values. Complications occurred in three patients after OLT, including miliary tuberculosis in one, graft rejection and retransplantation in one, and hepatic artery thrombosis in one. All patients survive with normal serum bilirubin level (follow up 6 to 116 months). Four have normal enzymes on post-OLT follow-up (30 to 95 months), follow a normal education program, and have a normal social life. One recently transplanted patient has progressively normalizing liver function tests 6 months after OLT. One patient transplanted at 8 y.o. (now 116 months post-OLT) has moderate neurological delay due to pretransplant kernicterus, and posttransplant chronic persistent hepatitis. Our series shows that OLT cures hyperbilirubinemia in CN patients, with an excellent survival prospect. The procedure should be decided upon before neurological sequelae occur, since these persist after transplantation. PMID- 7482715 TI - Timing, significance, and prognosis of late acute liver allograft rejection. AB - Liver allograft rejection is usually divided into acute (cellular) rejection and chronic (ductopenic) rejection. Most cases of acute rejection occur within four weeks of transplantation. There is a paucity of published literature on late acute rejection (LAR) in liver allografts and little is known about factors affecting its occurrence and outcome. To study the predisposing factors, clinical presentation, and prognosis of LAR, data prospectively collected on consecutive adult patients who underwent liver transplantation between 1982 and 1994, were analyzed. LAR was defined as histologically confirmed acute cellular rejection occurring 30 or more days after liver transplantation. Of the 717 patients, 59 (7.5%) had 71 episodes of LAR. Fifty-seven episodes were seen during the first year after transplantation, the remaining occurring between 1 and 6 years. Age, sex, pretransplant diagnosis, donor match of HLA, and blood groups was not associated with risk of LAR. Twenty-seven (38%) episodes were preceded by subtherapeutic blood levels of cyclosporine/FK506 (< 100 ng/ml and < 5 ng/ml, respectively) while an additional 6 (8%) had marginally low blood levels (< 150 ng/ml and < 10 ng/ml, respectively). Treatment with high-dose prednisolone resulted in complete resolution of rejection in 36 (51%) episodes, partial response in 21, and no response in 14 patients. Sixteen patients (27%) developed chronic rejection and graft loss. Development of chronic rejection was not affected by age or sex of the patient, timing of LAR, or histological severity of AR. Delayed response to therapy during an earlier episode of AR, and histological findings of centrilobular necrosis or bile duct loss at the time of diagnosis of LAR were associated with high risk of progression to chronic rejection and graft loss. PMID- 7482716 TI - Enhanced risk of steroid-resistant acute rejection following pretransplant steroid therapy in liver graft recipients. AB - Most episodes of acute rejection will resolve after steroid therapy without detrimental consequences on the liver allograft. However, steroid-resistant acute rejection may induce irreversible lesions of the graft and is associated with an increased risk of chronic rejection. The aim of this study was to determine whether there were predictive factors for steroid-resistant acute rejection after liver transplantation. A total of 108 adult liver recipients with a follow-up of at least 2 years have been analyzed; sixty-two (57%) patients had at least one episode of acute rejection. The rates of steroid resistance were 35%, 52% and 83% after a first (n = 62), second (n = 25), or third (n = 7) episode of acute rejection, respectively. Steroid resistance of acute rejection was significantly associated with a past history of pretransplant steroid therapy (P = 0.004). High levels of ALT (P = 0.03) and serum bilirubin (P = 0.002) were also predictive of steroid-resistant acute rejection. Eight (7%) patients eventually developed chronic rejection. Predictive factors for chronic rejection included steroid resistant acute rejection (P = 0.01), recurrent acute rejection (P = 0.03), and CMV infection (P = 0.01). In conclusion, this study suggests that pretransplant steroid administration or high levels of ALT and bilirubin in rejecting patients are risk factors for steroid resistance and should lead to aggressive antirejection therapy without delay. PMID- 7482717 TI - Prevention of hepatic artery thrombosis in pediatric liver transplantation. AB - Hepatic artery thrombosis after orthotopic liver transplantation is a serious complication, especially in children. We report our experience with intensive anticoagulant therapy during and after living-related liver transplantation in pediatric recipients. Twenty-four patients between 5 months and 15 years of age were studied. The mean diameter of the anastomosed hepatic arteries was 2.7 mm. The anticoagulant therapy consisted of low-molecular-weight heparin, antithrombin III concentrates, prostaglandin E1, fresh frozen plasma, and a protease inhibitor. The profiles of the coagulation and fibrinolytic systems were monitored by measuring several parameters, including plasma levels of thrombin antithrombin III complex, antithrombin III, plasmin-alpha 2 plasmin inhibitor complex, fibrin degradation product D-dimer, tissue type-plasminogen activator, and plasminogen activator inhibitor-1. Acceleration of the coagulation system and delayed recovery of the fibrinolytic system were observed during the early postoperative days. The plasma level of antithrombin III activity was maintained within the normal range by the administration of antithrombin III concentrates. None of the recipients developed hepatic artery thrombosis. Children have been reported to be at a greater risk of developing hepatic artery thrombosis than adults due to the small diameters of their hepatic arteries and the postoperative hypercoagulable state. We believe that the intensive anticoagulation therapy described in this study, the main concept of which is the early correction of imbalance between the coagulant and anticoagulant systems, could become a model for the prevention of hepatic artery thrombosis in pediatric liver transplantation patients. PMID- 7482718 TI - A genetic marker of high TNF-alpha production in heart transplant recipients. AB - The cytokine TNF-alpha has been implicated in the pathogenesis of both acute and chronic transplant rejection. Levels of the cytokine are known to vary in a normal population, leading to speculation that high responders may be at greater risk of rejection. Particular TNF region polymorphic markers have been associated with increased TNF-alpha levels and a biallelic polymorphism has been identified at position -308 of the TNF-alpha promoter that may contribute significantly to the interindividual variation in healthy persons. We describe here a new association between a polymorphic locus in the TNF gene region and increased production of TNF-alpha in heart transplant recipients. We studied two microsatellite markers that flank the TNFA gene, as well as a biallelic polymorphism at position -308 of the TNFA promoter, and found that the microsatellite allele TNFd3 was significantly associated with the capacity of leukocytes to produce TNF-alpha in vitro. No association was demonstrated for the promoter region polymorphism. Patients were receiving cyclosporine (CsA) and prednisolone (pred) at the time of sampling, which are known to interrupt 5' regulation of TNFA transcription in T cells and macrophages and may therefore negate the influence of the -308 polymorphism. Because of this we suggest that TNFd3 may be a marker for a 3' repressor region polymorphism that is of importance in immunosuppressed individuals. PMID- 7482719 TI - The relation of interleukin-6, tumor necrosis factor-alpha, IL-2, and IL-2 receptor levels to cellular rejection, allograft dysfunction, and clinical events early after cardiac transplantation. AB - Immunoregulatory cytokines have been implicated in the pathophysiology of graft dysfunction after heart transplantation (HTx). In 15 consecutive patients undergoing HTx we prospectively determined levels of interleukin-6 (IL-6), tumor necrosis-factor-alpha (TNF-alpha), interleukin-2 (IL-2), and soluble-interleukin 2-receptor (sIL-2-R) at eight points in time during biopsy and right heart catheterization and within 12 hr of echocardiography during the first three months after HTx. Blood was taken from the pulmonary arterial line. IL-6-levels correlated positively with hemodynamic and echocardiographic parameters of pump dysfunction--namely, pulmonary capillary wedge pressure, pulmonary arterial pressure, right atrial pressure, heart rate--and negatively with isovolumic relaxation time and stroke volume independent of the degree of cellular rejection as classified by the ISHLT criteria. A similar pattern was found for TNF-alpha- and sIL-2-R, while IL-2 correlated negatively with left and right heart filling pressures and positively with fractional shortening. In the three patients who died of sepsis or multiorgan failure within the study period IL-6-, TNF-alpha, and sIL-2-R-levels were elevated and IL-2-levels were suppressed compared with the 12 patients with a stable clinical course. IL-6 and sIL-2-R correlated positively while IL-6 and IL-2 correlated negatively. In this pilot study, a cytokine pattern with elevated levels of IL-6, TNF-alpha, and sIL-2-R as well as suppressed levels of IL-2 in the early period after HTx corresponds to impaired hemodynamics independent of cellular rejection and may indicate an unfavorable prognosis. These cytokines may therefore be useful for monitoring and warrant further study. PMID- 7482720 TI - Detection of donor-derived cells by polymerase chain reaction in neonatally tolerant mice. Microchimerism fails to predict tolerance. AB - Although it has long been appreciated that establishment of chimerism is important in the acquisition and maintenance of allograft tolerance, the importance of this relationship has been reemphasized recently. Using the exquisite sensitivity of the polymerase chain reaction we have studied qualitatively and quantitatively the presence of donor-derived chimeric cells in relation to the ability of neonatally injected mice to display skin graft tolerance or rejection. We have found that virtually all mice that receive neonatal injections of F1 hematopoietic cells acquire donor-derived chimerism that is detectable in blood, spleen, lymph nodes, and thymus. Surprisingly, neither the presence nor the quantity of chimeric cells predicts whether a particular neonatally injected mouse will accept or reject donor-specific skin allografts. Moreover, whether the test skin allograft is accepted (tolerance) or rejected by neonatally injected mice, chimerism typically remains detectable within recipient lymphoid tissues. In functionally tolerant mice, challenge with a test skin allograft actually leads to a remarkable expansion in donor-derived genetic sequences, implying that donor-derived cells have been induced by the graft to undergo proliferation. Since persistence of chimerism without proliferation after test grafting is characteristic of mice that fail to display tolerance, we believe that achievement of a threshold level of donor-derived alloantigen may be necessary to retain the tolerant state. We conclude that chimerism is essential for the induction of neonatally induced tolerance, and its expansion may play an important role in the maintenance of that tolerance, when challenged by an allograft. PMID- 7482721 TI - Graft-infiltrating T helper cells, CD45RC phenotype, and Th1/Th2-related cytokines in donor-specific transfusion-induced tolerance in adult rats. AB - Specific tolerance to LEW.1W (RT1u) heart allografts can be induced in adult LEW.1A (RT1a) rats by donor-specific blood transfusion (DST). We have previously shown that both rejected and tolerated grafts are heavily infiltrated by T lymphocytes, and that in both cases these T cells are capable of developing similar cytotoxic responses against donor cells in vitro; tolerance is therefore not due to the deletion of alloreactive T cells. At the same time, we found that the accumulation of IL-2 and IFN-gamma mRNA was decreased in tolerated grafts compared with rejected grafts. These results suggested that the induction of allograft tolerance in DST-treated animals could be mediated by anergy or suppression of graft-infiltrating Th1 cells. Although Th1 and Th2 clones have not yet been characterized in the rat, peripheral CD4+ rat T cells can be divided into two populations, based on their expression of the isoform RC of the CD45 molecule. Upon activation, CD45RChigh CD4+ T cells produce IL-2 and IFN-gamma and responsible for the induction of the graft-versus-host reaction, whereas CD45RClow CD4+ T cells produce IL-4 in vitro and provide B cell help. In the present study, we show that heart allografts from both DST-treated and untreated rats were infiltrated by equivalent numbers of leukocytes, of which CD4+ T cells also made up similar percentages. Among these CD4+ T cells, we observed that in allografts from DST-treated recipients the CD45RChigh population on day 5 was very significantly smaller (P = 0.004) than in the untreated group, while CD45RClow populations remained comparable. Moreover, using a new quantitative RT PCR method, we found a dramatic reduction in the accumulation of IL-2, IFN-gamma, IL-10, IL-4, and IL-13 mRNA in hearts from DST-treated recipients compared with those of untreated recipients during the week following transplantation. These results show that in heart allografts from DST-treated recipients, despite phenotypic changes suggesting Th1 inhibition by Th2 imbalance, T helper function was inhibited as a whole, and that in vivo the phenotype CD4+ CD45RClow does not always correlate with Th2-related cytokine-producing cells. PMID- 7482722 TI - Transplantation tolerance to rat cardiac and islet allografts by posttransplant intrathymic inoculation of soluble alloantigens. AB - The search for strategies for induction of specific tolerance in adult animals that will avoid long-term host immunosuppression with its complications has led to the deliberate introduction of alloantigens (Ag) into the adult thymus. However, pretransplant intrathymic (IT) inoculation of alloantigens (Ag), which has consistently induced tolerance to vascularized and neovascularized allografts in adult rodents, has limited future clinical application. To overcome the practical limitations of pretreatment, we have examined in the Lewis-to-WF combination the effect on graft survival of either simultaneous or posttransplant IT inoculation of soluble Ag obtained from 3M KCl extracts of donor T cells in transiently rabbit antirat lymphocyte serum (ALS) immunosuppressed recipients. While IT injection of 2.0 mg soluble Ag alone on day of cardiac transplantation caused acute graft rejection, IT inoculation of 2.0 mg Ag combined with 1 ml ALS transient immunosuppression of the recipient on day 0 led to long-term graft survival (> 250 days) in 5/6 recipients. Similarly, IT injection of soluble Ag on posttransplant day 3 or day 7 combined with 1 ml ALS on day 0 relative to allografting resulted in permanent graft survival in all recipients. In contrast, intravenous injection of soluble Ag combined with ALS immunosuppression on day 0 led to acute graft rejection that paralleled the rejection seen in ALS treated controls. Third-party Brown Norway (BN) hearts were acutely rejected in similarly prepared recipients of IT-Ag, thus confirming donor specificity. The long-term unresponsive Wistar-Furth (WF) recipients challenged 100 days after cardiac transplantation with a second-set graft specifically and permanently (> 100 days) accepted the second-set donor cardiac allografts, thus demonstrating donor specific tolerance. In similar experiments, IT inoculation of 2 mg soluble Ag combined with transient ALS immunosuppression resulted in donor-specific unresponsiveness to islets in the same rat combination of Lewis-to-WF. Our findings suggest that this new strategy of immunologic manipulation of the adult thymus offers a safe, effective, and reproducible method of inducing tolerance that may have therapeutic application in cadaveric organ transplantation. PMID- 7482723 TI - Mycophenolic acid, an inhibitor of IMP dehydrogenase that is also an immunosuppressive agent, suppresses the cytokine-induced nitric oxide production in mouse and rat vascular endothelial cells. AB - Mycophenolic acid (MPA), an inhibitor of IMP dehydrogenase and de novo GTP biosynthesis, also has immunosuppressive activity. The effect of MPA on nitric oxide (NO) production by rodent brain vascular endothelial cells in culture was investigated. MPA inhibited NO production by mouse and rat brain endothelial cells that had been stimulated with a combination of interferon-gamma and tumor necrosis factor-alpha. The 50% inhibitory concentration (EC50) was in the range of 0.5-1.0 microM. However, MPA had no effect on basal NO production in mouse brain vascular endothelial cells. Brequinar, an inhibitor of de novo pyrimidine synthesis, had no effect on NO production in cytokine stimulated endothelial cells. Guanosine, which can act as a salvage pathway precursor for GTP biosynthesis, reversed the inhibitory effect of MPA in a dose-dependent fashion. We suggest that inducible NO synthase activity is dependent on GTP level and can be blocked by curtailing IMP dehydrogenase activity. PMID- 7482724 TI - Protection of xenogeneic cardiac endothelium from human complement by expression of CD59 or DAF in transgenic mice. AB - We investigated the ability of membrane-bound human complement regulatory proteins to control complement-driven humoral immune reactions on murine microvasculature. The human complement regulatory proteins CD59 and DAF were expressed using heterologous promoters in a variety of tissues in transgenic mice. Animals expressing these gene products are healthy and exhibit significant levels of endothelial cell expression of CD59 and DAF in cardiac muscle. Transgenic hearts perfused with human plasma exhibited profound reductions in the level of complement deposition compared with nontransgenic controls. We have also produced transgenic pigs that express these two human genes. Our results indicate that expression of complement regulatory proteins can control activation of complement and suggest that these proteins may have therapeutic applications in some inflammatory diseases and in the development of xenogeneic organs for human transplantation. PMID- 7482725 TI - A synthetic dimeric HLA class I peptide inhibits T cell activity in vitro and prolongs allogeneic heart graft survival in a mouse model. AB - A peptide derived from the alpha 1 domain of the human HLA class I heavy chain (amino acids 75-84; B2702.75-84) has been shown to inhibit human cytotoxic T and NK cell activity in a non-allele-restricted manner. In vivo, this peptide prolonged skin allograft survival in a murine model. Here we demonstrate prolongation of heart allograft survival in mice and extend the characterization of the immunomodulatory activity of B2702.75-84. Similar to what has been observed with retrovirus-derived peptides, the inhibitory capability of this peptide was increased when bound to a carrier protein. An increased immunomodulatory activity was also observed with the dimeric peptide B2702.84-75 75-84 or the multimeric B2702.75-84.MAP. This peptide not only inhibited cytotoxic T and NK cells but also anti-CD3-induced T cell proliferation as well as a mixed lymphocyte reaction (MLR). Flow cytometric analysis of T cells harvested from anti-CD3-stimulated spleen cell culture in the presence of B2702.84-75-75-84 showed decreased expression of activation markers (CD25, ICAM 1, Pgp-1, CD69) compared with untreated control cultures. The superior activity of B2702.84-75-75-84 could also be demonstrated in vivo. Administration of B2702.84-75-75-84 prolonged the survival of B6 (H2b) hearts in CBA (H2k) recipients to 15 +/- 2.7 (P = 0.0002 vs. control) days compared with 11.4 +/- 2.6 (P = 0.01) days in B2702.75-84 treated animals and 7.5 +/- 1.1 days in untreated controls. Administration of control peptides had no significant effect on allograft survival. In combination with a subtherapeutic dose of cyclosporine, B2702.75-84 induced long-term graft survival in 60% of recipients. PMID- 7482726 TI - Donor antigen-presenting cell-independent rejection of islet xenografts. AB - Donor-derived antigen-presenting cells (APC) are thought to serve as major stimulators for triggering the rejection of tissue allografts. However, the capacity of APC to stimulate xenogeneic T cells is generally deficient relative to the corresponding response from allogeneic T cells. For this reason, the contribution of donor-type APC to xenogeneic graft rejection remains unclear. Using a concordant species combination (rat to mouse), we examined the requirement for donor-type APC in triggering islet xenograft rejection. While the depletion of donor-type APC resulted in indefinite allograft survival, similar depletion of APC from xenogeneic rat islets resulted in only modest graft prolongation. Furthermore, APC-depleted rat xenografts were rejected by a CD8+ T cell-independent mechanism, as determined by appropriate depletion of T cell subsets through monoclonal antibody therapy. This contrasts with the dependence of islet allograft rejection on both CD4+ and CD8+ T cells. Although in vitro experiments show that rat APC can directly stimulate mouse T cells, rat APC do not appear to be required for xenograft immunity in vivo. We conclude that the mechanisms of islet allograft and xenograft rejection differ both in the dependence on donor-type APC and in the role of T cell subsets in the response. PMID- 7482727 TI - Inhibition of transplant rejection following treatment with anti-B7-2 and anti-B7 1 antibodies. AB - Antigen-specific T cell activation depends initially on the interaction of the T cell receptor (TCR) with peptide/MHC. In addition, a costimulatory signal, mediated by distinct cell surface accessory molecules, is required for complete T cell activation leading to lymphokine production and proliferation. CD28 has been implicated as the major receptor on T cells responsible for delivering the costimulatory signal. Although two distinct ligands for CD28, B7-1 and B7-2, have been identified on antigen-presenting cells (APC), the co-stimulatory role of each molecule during a physiological immune response remains unresolved. In the present study, the relative roles of B7-1 and B7-2 interactions were evaluated in an allogeneic pancreatic islet transplant setting. In isolation, anti-B7-2 mAbs and, to a much lesser degree, anti-B7-1 mAbs suppressed T cell proliferative responses to allogeneic islets or splenic APC in vitro. Maximal inhibition of the allogeneic response was observed using a combination of the anti-B7-1 and anti-B7 2 mAbs. Administration of anti-B7-2 but not anti-B7-1 mAbs prolonged C3H allograft survival in B6 recipients, with a combination of both mAbs significantly prolonging rejection beyond either mAb alone. The immunosuppressive effects of the in vivo mAb treatment were not manifested in in vitro analyses as T cells isolated from suppressed mice responded normally to allogeneic stimuli in terms of both proliferation and lymphokine production. However, combined mAb therapy in vivo selectively delayed CD4+ T lymphocyte infiltration into the graft. These data suggest that both B7-1 and B7-2 costimulatory molecules are active in vivo, although B7-2 plays a clearly dominant role in this allograft model. The mechanism of immune suppression in vivo remains unresolved but may occur at sites distinct from the allograft. PMID- 7482728 TI - The use of non-heart-beating cadaver donors in experimental liver transplantation. PMID- 7482729 TI - Successful transplantation of kidneys with hemosiderosis. PMID- 7482730 TI - Reversal of steroid-resistant rejection in renal allograft recipients using FK506. PMID- 7482731 TI - Successful orthotopic liver transplantation for fulminant liver failure in a child with autosomal recessive chronic granulomatous disease. PMID- 7482732 TI - Procurement and preparation of human isolated small intestinal grafts for transplantation. AB - We have developed a donor operation that incorporates en bloc removal of the liver and intestine with a limited surgical resection in vivo. Over the past 18 months, we have used the following technique for the retrieval and preparation of seven isolated small intestinal allografts. The donor operation and bench preparation can be divided into three phases. During the first phase, the small intestine is removed with the liver, pancreas, and an aortic segment. In the second phase performed ex vivo, the donor liver can be separated from the specimen. The third phase involves additional bench dissection to yield an isolated intestinal allograft. The principle advantage of this technique is that it reduces potential liver injury by minimizing the surgical dissection required in vivo. Also, dividing the liver from the intestine ex vivo allows the organs to be separated in a bloodless field under controlled conditions that may be especially important when two different surgical teams are involved. PMID- 7482734 TI - Impact of racial genetic polymorphism on the probability of finding an HLA matched donor. AB - As successful organ or marrow transplantation correlates with the degree of HLA compatibility between patient and donor, registries have been developed to facilitate matching. However, racial minority groups have a lower chance of finding a match. We evaluate the impact of the biology of racial genetic polymorphism upon the probability of finding an HLA match for patients of different racial groups. The National Marrow Donor Program has compiled the HLA types of 20,449 patients and 1,625,159 potential volunteer donors. These HLA types were used to estimate the probability of finding an HLA-matched donor for patients of different racial groups. We estimated the HLA haplotype frequencies for different races, and then determined the probability of finding matched donors, given several hypothetical registry sizes. We confirmed that patients of minority races searching the current National Marrow Donor Program registry have low probabilities of finding matches. This was only partly due to the smaller number of donors from these racial minorities, as the observation persisted even when hypothetical donor registry sizes were the same for all racial groups. We demonstrate that African-Americans are more polymorphic with respect to HLA, and are hence less likely to find donors at any given registry size. An increase in the recruitment of minority racial groups for organ and marrow donors will only partially alleviate the problem of equal access to HLA matches for patients belonging to racial minority groups. It will therefore be important to attempt to improve methods for transplantation using HLA-mismatched donors. PMID- 7482733 TI - Protective effects of an indenoindole antioxidant on coronary endothelial function after long-term storage. AB - Experiments were designed to evaluate function of the endothelium and smooth muscle of coronary arteries following storage of hearts in cardioplegia containing an inhibitor of lipid peroxidation (H 290/51, cis-7-methyl-9-methoxy 5,5a, 6,10b tetrahydroindeno [2,1-b] indole). Canine hearts were perfused with crystalloid cardioplegia (Plegisol, 15 ml/kg, 4 degrees C) and left circumflex arteries were isolated and studied either immediately (group I, n = 6), or after storage of the hearts at 4 degrees C for 10 (group II, n = 6) or 24 hr with (group III, n = 6) or without (group IV, n = 6) addition of H 290/51. The final concentration of H 290/51 was 1 mumol/L. Arteries were removed, cut into rings, and suspended in organ chambers for measurements of isometric force. In selected rings, the endothelium was removed in order to study the function of the smooth muscle. In order to discriminate effects of ischemia/reperfusion and protective properties on coronary endothelium or smooth muscle, drugs with different mechanisms were used. The function of the endothelium were studied with the alpha 2-adrenergic agonist UK 14,304, bradykinin and A 23187. The smooth muscle function were studied with isoproterenol and nitric oxide. Endothelium-dependent relaxations to the alpha 2-adrenergic agonist UK 14,304 and bradykinin, but not to A 23187, were reduced significantly in arteries from hearts stored for 24 hr in cardioplegic solution alone. Relaxations of arteries from hearts stored for 24 hr with H 290/51 were comparable to those arteries from hearts that were not stored. Endothelium-independent relaxations to isoproterenol and nitric oxide among the different groups were comparable. These results suggest that storage of canine hearts with crystalloid cardioplegia selectively inhibits endothelium dependent relaxations mediated by receptor activation. Inhibition of lipid peroxidation with H 290/51 preserves these relaxations and may therefore represent a therapeutic alternative to preserve hearts used for transplantation. PMID- 7482735 TI - The fate of renal allografts functioning for a minimum of 20 years (level 5A)- indefinite success or beginning of the end? A proposed classification of long term allograft survivals. AB - Fifty-five renal allografts (44 from living-related and 11 from cadaver donors) that have functioned for at least 20 years (mean 22.9 +/- 2.3, range 20.1 to 30.7 years) were evaluated in three groups based on renal function: group I (n = 26), with a GFR of > or = 60 ml/min/1.73 m2 or serum creatinine < or = 1.4 mg/dl and no proteinuria; group II (n = 9), with a GFR of > or = 60 ml/min/1.73 m2 or serum creatinine < or = 1.4 mg/dl but > 150 mg proteinuria/24 hr; and group III (n = 20), with a GFR < 60 ml/min/1.73 m2 and/or serum creatinine > 1.4 mg/dL with or without proteinuria. Allograft factors, including acute rejection (AR) in 62% (34/55) and delayed function (DF) in 55% (6/11) of the cadaver grafts, did not preclude 20-year success and the prospect of continued survival since they were not significantly more frequent in group I, II, or III. However, AR was confined to a limited period within the first three months posttransplant in 18/18 recipients in groups I and II but only in 7/16 of group III (P = 0.0002). In groups I and II AR was treated with IVMP in 14/18 cases and only 6/16 in group III (P = 0.035). Donor age < or = 50 years and recipient age < or = 40 years each occurred in 87% (48/55) of these transplants. One- or two-HLA haplotype matching was present in 98% (43/44) of living related transplants. Major risks to the recipient were coronary artery disease (11 cases and 3 deaths), malignancy (18 cases and 1 death), and severe infection and hepatitis (35 cases and 3 deaths, 2 of whom also had coronary artery disease). Hypertension occurred in 25 recipients and diabetes mellitus in 12. Potential open-end success was compromised by renal dysfunction in groups II and III, but appeared possible in 12 of the 26 patients in group I. There is no apparent "safe-haven" point of time for immunosuppressed renal allograft recipients, who remain at increased risk for eventual renal allograft dysfunction, as well as cardiovascular, neoplastic, infectious, and metabolic diseases. In order to clarify and standardize the words "long-term," a simple classification of long-term allograft survivals is proposed. PMID- 7482736 TI - Correlation between donor age and the pattern of liver graft recovery after transplantation. AB - We have observed an increased rate of delayed nonfunction (DNF) of liver grafts procured from older donors. The aim of this study was to correlate donor age and the patterns of graft failure after transplantation. Pattern of liver injury, synthetic function, and graft survival in recipients receiving liver grafts from donor older than age 50 (group I, n = 95) were compared with matched cohort of recipients transplanted with grafts from donors age 20-30 (group III, n = 50). Primary nonfunction (PNF) of the graft was defined as non-recoverable hepatocellular function necessitating emergency retransplantation within 72 hr. DNF was defined as marginal graft function necessitating retransplantation within one month. Recipient characteristics, including age and preoperative UNOS status, were similar between groups. Ischemic/reperfusion injury, reflected by SGOT and SGPT was more severe in older donors. PNF occurred at similar frequencies for all groups (7%). Normal liver function was regained in 76% of recipients in group I, and in 92% in group II. However, cholestatic pattern was observed in recipient of grafts from group I donors. Rapid rise in bilirubin, despite normalization of prothrombin time and liver transaminases, was the hallmark of DNF. DNF resulted in higher retransplantation rate in group I (24% vs. 8% in group II). Donor age did not affect patient survival. Liberalizing criteria for donor selection, and acceptance of older donors is a calculated risk. Over 75% of the recipients will regain normal liver function. However, a higher number of these grafts will exhibit slow recovery after transplantation, and a significant rate of DNF. Recognition of such pattern and early retransplantation should decrease mortality. PMID- 7482737 TI - Apoptosis in the human allografted kidney. Analysis by terminal deoxynucleotidyl transferase-mediated DUTP-botin nick end labeling. AB - Apoptosis is a distinct form of cell death and occurs under a variety of physiological and pathological conditions. This study was conducted to examine the occurrence of apoptotic cell death in 44 formalin-fixed, paraffin-embedded biopsy specimens from allografted kidneys by the terminal deoxynucleiotidyl transferase (TdT)-mediated D-UTP-biotin nick end labeling (TUNEL) method. Careful observation of routine hematoxylin and eosin sections revealed a few apoptotic cells in cortical tubules. TUNEL signal was detected variably in tubular epithelia, and occasionally in lymphocytic cells and endothelium. The number of tubular epithelia demonstrating TUNEL signals was highest for cyclosporine tubulopathy, followed by acute rejection of very mild or mild grade, and then by chronic allograft nephropathy. Protocol biopsies from normally functioning grafts showed the least apoptotic cells, with the number being significantly lower than that of both cyclosporine tubulopathy (P < 0.01) and acute rejection (P < 0.05). Two specimens of acute accelerated rejection with diffuse hemorrhagic necrosis showed nonspecific signals in a few epithelia. These findings suggest that acute rejection or cyclosporine nephropathy not infrequently induces apoptosis of tubular epithelia, which might lead to tubular atrophy or loss, resulting in chronic transplant nephropathy. PMID- 7482738 TI - The impact of pretransplantation hepatitis C infection on the outcome of renal transplantation. AB - Controversy exists regarding the impact of pretransplantation HCV infection on the outcome of renal transplantation. We compared the prevalence of posttransplantation liver disease and graft and patient survival among kidney transplant recipients with and without anti-HCV at the time of transplantation. Pretransplantation sera from 103 randomly selected recipients of kidneys from anti-HCV-negative donors were tested for anti-HCV using a second generation ELISA. Twenty-three (22%) were positive for anti-HCV and 80 (78%) were negative. Anti-HCV-positive recipients had a longer time on dialysis (P = 0.003) and had a higher number of previous transplants (P = 0.01). Further, 61% of anti-HCV positive patients had a history of liver disease compared with 13% of anti-HCV negative patients (P < 0.001). HCV RNA was detected in the pretransplantation serum in 61% of anti-HCV positive recipients compared with 5% of anti-HCV negative recipients (P < 0.001). Clinical follow-up on both anti-HCV-positive and -negative patients was obtained until December, 1993. Median posttransplantation follow-up among recipients with anti-HCV prior to transplantation (45 months) was shorter (P = 0.05) than that for recipients without anti-HCV (66 months). For recipients with anti-HCV prior to transplantation, the relative risk of posttransplantation liver disease was 5.0 (95% confidence intervals of 2.4 to 10.5); the relative risk of graft loss was 1.3 (95% confidence intervals of 0.6 to 2.6); the relative risk of death was 3.3 (95% confidence intervals of 1.4 to 7.9), and the relative risk of death due to sepsis was 9.9 (95% confidence intervals of 2.6 to 38.3). The results of this study demonstrate that pretransplantation HCV infection is associated with an increased risk of liver disease and death after renal transplantation. These results raise the question of whether anti-HCV-positive patients on dialysis should be offered renal transplantation as opposed to continuing dialysis. PMID- 7482739 TI - Tolerance induction to cardiac allografts by simultaneous or sequential intrathymic inoculation of disparate alloantigens. AB - Recent studies show that the adult immune system can be manipulated to accept allografts by the intrathymic (IT) inoculation of donor alloantigens (Ag). To make this model clinically applicable to cadaveric transplants, we have examined the effect of simultaneous or sequential IT injection of two disparate Ags on graft survival. In initial experiments, Wistar Furth (WF) rats received IT injections of a single or an admixture of 2 x 10(7) Lewis T cells and 2 x 10(7) BN T cells 7 days prior to cardiac transplantation. Untreated WF recipients acutely rejected single Lewis, BN, and ACI heart allografts, respectively, at times equivalent to rejection of double hearts while IT injection of single or double 2 x 10(7) Lewis and 2 x 10(7) BN T cells on day -7 also led to acute graft rejection. As expected, IT injection of donor-type resting T cells combined with 1 ml ALS recipient immunosuppression on day -7 led to donor-specific Lewis or BN permanent graft survival (> 200 days). The long-term unresponsive WF recipients challenged 100 days after cardiac transplantation with 2nd-set grafts specifically and permanently accepted 2nd-set donor cardiac allografts. IT injection of an admixture of 2 x 10(7) Lewis and 2 x 10(7) BN T-cells combined with 1 ml ALS on day -7 resulted in 100% permanent double (Lewis and BN) allograft survivals (> 150 days). Similar treatment led to acute rejection of third-party (ACI) grafts while the simultaneously transplanted Lewis hearts survived indefinitely (> 150 days). In the second group of experiments, unresponsive recipients of single (Lewis or BN) cardiac allografts were given IT inoculation of a second Ag in a sequential fashion 30 or 100 days after primary heart transplant. Unresponsive WF recipients of Lewis grafts accepted permanently (> 100 days) second BN grafts that were transplanted 30 or 100 days after primary Lewis graft and 7 days after IT injection of BN T-cells and 1 ml ALS. Similarly, unresponsive WF recipients of BN grafts accepted second Lewis grafts after IT injection of Lewis T cells combined with ALS, 30 or 100 days after primary BN grafts. To define the mechanism of peripheral tolerance in this model, we have shown that adoptive transfer of unseparated spleen cells from unresponsive recipients induces prolonged double (Lewis and BN) graft survivals (> 60 days) but not ACI grafts in sublethally irradiated (400 rads) syngeneic (WF) recipients, thus suggesting that maintenance of tolerance is, in part, dependent on suppressor/regulatory cell network.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 7482740 TI - Nutritional immunomodulation enhances cardiac allograft survival in rats treated with donor-specific transfusion and cyclosporine. AB - The aim of this study was to assess the efficacy of an enteral diet fortified with arginine, RNA, and fish oil (Impact), alone and in combination with cyclosporine (CsA) and donor-specific transfusion (DST) on allograft survival in the ACI:Lewis rat cardiac transplant model. Animals were fed ad libitum with either standard rat chow or Impact diet. Six groups were studied; these consisted of untreated recipients fed either standard diet or Impact diet; recipients treated with CsA 10 mg/kg on the day prior to engraftment (day-1) followed by 2.5 mg/kg/d, day 0-->day+13 and fed with either standard diet or Impact; and animals given a DST (1 ml) on day-1, CsA as described previously and fed either standard diet or Impact. Untreated animals standard diet (group 1, n = 8) rejected their allografts at 7.0 +/- 0.0 days, while those fed Impact (group 2, n = 9) had graft survival of 12.8 +/- 2.1 days, (P = .01 versus group 1). Animals treated with CsA alone and standard diet (group 3, n = 9) rejected at 30.3 +/- 4.8 days, while the combination of CsA and Impact diet (group 4, n = 8) rejected at 33.0 +/- 9.5 days -minimally improved survival compared with group 3. Animals treated with DST/CsA and standard diet (group 5, n = 7) rejected at 72.1 +/- 6.8 days, while the substitution of Impact for standard diet (group 6, n = 8) led to a significant graft prolongation to 275 +/- 53 days, n = 8 (P < .015 vs. groups 1-5). These data suggest that Impact diet alone can have potent immunomodulatory properties but may require the addition of DST/CsA to realize its potential. These findings underscore the potential of dietary immunomodulatory therapy to prevent rejection and promote tolerance to solid organ allografts. PMID- 7482741 TI - Mechanisms of alloimmunization and immunosuppression by blood transfusions in an inbred rodent model. AB - During refrigerated storage leukocytes in donor blood progressively undergo apoptosis followed by secondary necrosis. Using an inbred rodent transfusion model, recipient animals received viable, necrotic, or apoptotic cells. While transfusion of viable blood MNCs stimulated production of IgM, IgG1 (Th2 type) and IgG2a (Th1-type) antidonor antibodies, leading to a suppression of subsequent DTH to donor antigens, transfusion of apoptotic donor cells led to neither alloimmunization nor immunosuppression. On the other hand transfusion of lysed donor cells resulted in production of IgM and IgG1 (Th2-type) antidonor antibodies and to a strong suppression of subsequent DTH to donor antigens. Intravenously administered spleen cells that had been depleted of professional APCs and enriched for B cells stimulated IgM antidonor antibodies but not IgG antibodies. Transfusion of such cells also led to suppression of subsequent DTH to donor antigens, probably through induction of anergy or apoptosis in alloantigen-reactive recipient cells. Depending on the duration of blood storage any or all of these 4 classes of cells may be present and Th2 and/or Th1 effector mechanisms can be generated following blood transfusion. PMID- 7482742 TI - The effect of human IL-2-activated natural killer and T cells on graft-versus host disease and graft-versus-leukemia in SCID mice bearing human leukemic cells. AB - We have previously reported that xenogeneic lethal acute graft-versus-host disease (GVHD) was induced by transplantation of a mixture of human IL-2 activated natural killer (NK) and T cells into SCID mice conditioned with 4 Gy total-body irradiation (TBI), but not by IL-2-activated pure human T cells or NK cells. TBI and transplantation of the mixture of activated cells were both required to produce the lethal effect. We now report the effect of human IL-2 activated NK, T, or NK+T effector cells on the development of acute and chronic GVHD and GVL in SCID mice bearing human leukemic cells. Ten days after being inoculated i.v. with 2 x 10(7) human U-937 or K-562 leukemic cells, SCID mice, hereafter termed hu-leukemic mice, were radiated with 4 Gy TBI and transplanted i.v. with 5 x 10(7) human IL-2-activated NK, T, or NK+T effector cells. Hu leukemic control mice received neither TBI nor effector cell transplantation. Acute GVHD-positive control SCID mice were transplanted with 5 x 10(7) H-2 incompatible C57Bl/6 splenocytes following 4 Gy TBI. The mice were observed for signs of GVHD and leukemia for 90 days. Twenty of 20 non-effector cell transplanted control hu-leukemic mice developed signs related to leukemia and died with leukemic infiltration in the brain, liver, kidney, and lung 50-65 days after inoculation. Flow cytometry (FC) demonstrated 21-89% human leukemic cell infiltration in the bone marrow. Fourteen of 14 hu-leukemic mice transplanted with NK+T effector cells did not develop signs of advanced leukemia but died within 17 days of acute GVHD. FC demonstrated no human leukemic cells in their marrow. Twelve of 15 and 18 of 25 hu-leukemic mice transplanted with either NK or T cells survived 90 days without any evidence of symptomatic leukemia (P < 0.01 compared with non-effector cell-transplanted groups). NK-transplanted hu-leukemic animals experienced mild-to-moderate acute GVHD during the first 10-20 days posttransplantation, but gradually recovered and did not develop chronic GVHD. Hu leukemic animals transplanted with T effector cells manifested no signs of leukemia or acute GVHD but chronic GVHD skin lesions appeared 80-90 days after transplantation. We conclude that acute GVHD, chronic GVHD, and GVL are associated but separable phenomena. PMID- 7482744 TI - Separation of feline bone marrow cells by counterflow centrifugal elutriation. Identification and isolation of presumptive early and late myeloid/erythroid progenitors. AB - Counterflow centrifugal elutriation (CCE) has been used to separate nucleated cells from mammalian bone marrow on the basis of size with the resultant isolation of hematopoietic cells in varying stages of lineage development. We examined the feasibility of identifying and isolating such cells from feline bone marrow. CCE was performed with a Beckman J6MI centrifuge and a Sanderson chamber, using a fixed rotor speed of 3000 rpm and collection of cells at (1) 16-, (2) 21 , (3) 25-, (4) 32 ml/min, and (5) a rotor off fraction. Recovery of the total input cells in four replicate experiments averaged 86%, with the maximum number of recovered cells in fraction 4. Analysis by flow cytometry and monoclonal antibodies revealed mononuclear cells in fractions 1 and 2 and early and late differentiating myeloid/erythroid cells in fractions 2 through 5. T lymphocytes and alloreactivity in a mixed lymphocyte reaction (MLR) were restricted to fractions 1 and 2; removal of T cells and MLR activity was accomplished by immunomagnetic depletion. In vitro cultures for clonogenic cells revealed CFU-GM and BFU-E colonies in fractions 2 through 5, with fraction 4 containing the greatest absolute number of myeloid colonies and fractions 3 and 4 the majority of the erythroid colonies. More important, in examining the plating efficiency for clonogenic cells in the different fractions it was found that this increased significantly in fractions 2 and 3 when the culture time was extended from 7 to 14 days; in contrast, fractions 4 and 5 reached their maximum plating efficiency within 7 days with no further increase on day 14. We interpret these findings to indicate the presence of late differentiating progenitors in the large-cell size fractions 4 and 5, while the smaller mononuclear cells in fractions 2 and 3 represent an earlier, more primitive population of hematopoietic cells requiring an extended time in culture for full colony development. PMID- 7482743 TI - Comparison of in vivo efficacy and mechanism of action of antimurine monoclonal antibodies directed against TCR alpha beta (H57-597) and CD3 (145-2C11). AB - Monoclonal antibodies (mAbs) directed against the T cell receptor (TCR) associated CD3 chains and against the TCR-alpha beta heterodimer can inhibit allograft rejection in humans and in experimental animals. Since the effects of stimulation through these cell surface structures may differ, it has been suggested that there could be advantages to targeting one structure versus the other. In order to directly compare two such mAbs for in vivo immunosuppressive properties and mechanisms of action, C57BL/10 mice were treated with mAbs H57-597 (H57, anti-alpha beta) or 145-2C11 (2C11, anti-CD3), either as intact mAb or as F(ab')2 fragments. F(ab')2 fragments of both mAbs had similar effects. Both prolonged skin allograft survival, preferentially depleted CD4+ T cells, downregulated IL-2 secretion, and failed to inhibit CTL. In contrast, the effects of the intact form of the two mAbs differed significantly. Intact H57 was far more effective than 2C11 in prolonging skin allograft survival and in inhibiting cytokine secretion and CTL function. This increased immunosuppressive effect was associated with a significantly more complete and prolonged depletion of both CD4+ and CD8+ T cells and down-modulation of TCR expression on remaining T cells. A markedly greater half-life was observed for H57, associated with reduced immunogenicity. These data suggest that the increased immunosuppressive properties of H57 are due to its reduced immunogenicity, rather than to differences in signal transduction, and support the argument that reducing the immunogenicity of mAbs in the clinical setting by "humanization" may result in improved efficacy. PMID- 7482745 TI - Enzymatic removal of alpha-galactosyl epitopes from porcine endothelial cells diminishes the cytotoxic effect of natural antibodies. AB - Xenotransplantation of tissues between discordant species such as pig into human is not yet feasible due to the problem of hyperacute rejection. This rapid response to xenogeneic tissue is mediated by natural antibodies that react with antigens on the xenograft. A number of xenoantigens consist of carbohydrate residues, and a terminal galactose in alpha linkage has been shown to be involved in hyperacute rejection of pig-to-human xenografts. We show that alpha-linked galactose on porcine endothelial cells is a major epitope recognized by IgG and IgM antibodies present in monkey and human sera. Endothelial cells that had been treated with alpha-galactosidase did not react with fluorescein-labeled Griffonia simplicifolia I B4 (GS-IB4), a lectin that detects the alpha-galactosyl epitope on intact cells. The reactivity of both human and cynomolgus monkey serum with endothelial cells was decreased by 59% to 90% after treatment with coffee bean alpha-galactosidase. Using a colorimetric assay for cell viability, we show that natural antibodies present in the sera of cynomolgus monkey and humans are cytotoxic to porcine endothelial cells in the presence of exogenously added complement. When the terminal alpha-galactosy residues were removed by enzymatic digestion, the cytotoxic effect of natural antibodies on porcine endothelial cells was diminished by > 80%. Evaluation of the time course of reappearance of the alpha-galactosyl epitope at the cell surface revealed that 48 hr after alpha galactosidase treatment, binding of GS-IB4 was diminished by 60%. These results suggest that glycosidase treatment of cells to be transplanted could prevent hyperacute rejection mediated by natural antibodies. PMID- 7482746 TI - The effect of aglycosylation on the immunogenicity of a humanized therapeutic CD3 monoclonal antibody. AB - The factors affecting the immunogenicity of a humanized gamma 1 CD3 monoclonal antibody (mAb) were investigated in transgenic mice that express the human CD3 antigen epsilon polypeptide (the mAb target antigen). Two derivatives of the mAb were employed, one with a normal, glycosylated Fc region (gamma 1 CD3 mAb), and the other with an aglycosylated Fc region (aglycosyl gamma 1 CD3 mAb). Comparisons of the antiglobulin responses elicited by the two derivatives in transgenic and nontransgenic mice demonstrated that Fab-mediated cell binding activity, dependent on target antigen expression, was a major positive determinant of CD3 mAb immunogenicity. A second positive factor was mAb Fc region glycosylation. At low dose levels the gamma 1 CD3 mAb consistently produced a higher antiglobulin response than the aglycosyl gamma 1 CD3 mAb. This was probably a result of the nonspecific, in vivo T cell activating property of the gamma 1 CD3 mAb, a consequence of its ability to cross-link T cells to Fc gamma receptor-bearing cells. (The aglycosyl gamma 1 CD3 mAb has a reduced Fc binding affinity for Fc gamma receptors and so does not activate T cells in vivo.) In support of this hypothesis, the gamma 1 CD3 mAb was able to nonspecifically enhance humoral immunity to an unrelated, coadministered antigen, whereas the aglycosyl gamma 1 CD3 mAb was not. The lower immunogenicity of the aglycosyl gamma 1 CD3 mAb correlated with a longer in vivo half-life and an improved capacity to block the target CD3 antigen. These results suggest that, as well as reducing the cytokine-induced side effects normally associated with CD3 mAb therapy, the nonactivating aglycosyl gamma 1 CD3 mAb will be less likely than the activating gamma 1 CD3 mAb to stimulate a neutralizing antiglobulin response. PMID- 7482747 TI - Functional studies of rat, porcine, and human pancreatic islets cultured in ten commercially available media. AB - There have been no extensive studies investigating the effect of tissue culture media on the in vitro functional characteristics of rat, porcine and human Islets of Langerhans. We therefore aimed to compare ten commercially available tissue culture media on the basis of their ability to maintain islet viability. Following isolation, islets were cultured free-floating in the ten media (RPMI 1640-11mM glucose (control), RPMI 1640-2.2mM glucose, Dulbecco's MEM, TCM 199, CMRL 1066, Iscove's MEM, Waymouth's MEM, Serum-Free medium, Ex-cell 300, Ham's F 12) and viability was assessed after 24 hr, 3 days, and 7 days on the basis of macroscopic appearance, cell membrane integrity, and insulin secretion in response to glucose stimulation both by dynamic incubation and by perifusion. Each islet species demonstrated physiological insulin release characteristics in all media--however, it was possible to distinguish between the media by comparing the stimulation indices calculated from the insulin release studies. Significantly higher stimulation indices were produced in Iscove's MEM for rat islets, in Ham's F-12 for porcine islets and in CMRL 1066 for human islets. Over the entire culture period a significant deterioration in function was observed in all species cultured in the control media, although this was reversed when islets were cultured in the optimal media. Furthermore, in the case of porcine and human islets a significant improvement in function over the seven-day period was noted in the optimal media. In conclusion, of the commercially available media, the optimal tissue culture medium for rat islets is Iscove's MEM, for porcine islets is Ham's F-12, and for human islets is CMRL 1066. PMID- 7482748 TI - The humoral immune response in humans following cross-perfusion of porcine organs. AB - A major question in xenotransplantation is the nature of the humoral response that would occur following the transplantation of a xenogeneic organ into an immunosuppressed recipient as such a response could mediate delayed types of injury to the graft. To begin to address this issue we characterized the changes in the properties of xenoreactive antibodies occurring in patients exposed to porcine organs under conditions simulating transplantation. In two patients whose blood had been cross-perfused through porcine livers as a treatment for hepatic failure, the titer of xenoreactive IgM increased by four-fold and the titer of xenoreactive IgG increased by sixty-fold within ten days after perfusion procedures. The xenoreactive IgM and IgG antibodies were specific for Gal alpha 1 3Gal based on binding to porcine endothelial cells and bovine thyroglobulin, which express this determinant, and on the decrease in binding following treatment of porcine endothelial cells or bovine thyroglobulin with alpha galactosidase. The sequential addition to endothelial cells of amounts of serum known to saturate antibody-binding sites obtained before and ten days after perfusion of porcine organs revealed no increase in binding of IgM above the level observed with serum obtained before perfusion, suggesting that new determinants were not identified. Moreover, the functional avidity of binding to porcine endothelial cells of IgM in serum obtained before and ten days after perfusion of porcine organs was unchanged. Even at later times, the presence of newly elicited antibodies against porcine aortic endothelial cell targets was not detected. Thus, exposure to porcine antigens in a vascularized organ results in increases in the levels of xenoreactive IgM and IgG antibodies--however, these antibodies exhibit properties similar to natural antibodies. PMID- 7482749 TI - Heterogeneity of HLA-B35. Oligotyping and direct sequencing for B35 subtypes reveals a high mismatching rate in B35 serologically compatible kidney and bone marrow donor/recipient pairs. AB - We used a simple HLA-B35 PCR/SSO-oligotyping procedure, combined with exon 3 direct sequencing for the analysis of B35 subtype frequencies in our population, and for the evaluation of the degree of B35-subtype compatibility in serologically matched unrelated bone marrow and kidney transplant pairs. B*3501 was the most frequent allele (0.6), followed by B*3503 (0.19), B*3502 (0.13), B*3508 (0.07), and B*3505 (< 0.01). HLA-B35-subtype matching of donors and recipients was strongly dependent on the stringency of ABDRB1 matching. Among 10 kidney donor/recipient pairs, only 30% were B35-subtype-matched. Due to the lack of ABDRB1 haplotype matching, this low degree of matching was not better than what would be expected on the basis of the subtype frequency distribution in the population. In contrast, HLA-B35 subtype compatibility was higher in unrelated bone marrow donor/recipient pairs that were serologically ABDR-matched: 30 of the 62 (48.4%) B35-positive combinations tested were B35-subtype-compatible. When all patient/donor plus donor/donor combinations (n = 160) were taken into account, 46% of the ABDR-matched pairs were B35-subtype-compatible. When only pairs that were DRB1/DRB3/DRB5-subtype-matched by oligotyping (n = 62) were considered, 71% were B35-subtype-compatible. The fact that a significant percent of patient/donor pairs matched by conventional HLA-typing are found incompatible, as shown here for B35, explains the difficulties in assessing the beneficial effect of HLA matching in transplantation. PMID- 7482750 TI - Cyclosporine-related leukoencephalopathy and PML in a liver transplant recipient. PMID- 7482751 TI - Arterialization of the portal vein in orthotopic and auxiliary liver transplantation. A report of three cases. PMID- 7482752 TI - Human ehrlichiosis in a liver transplant recipient. PMID- 7482753 TI - The use of the recipient sigmoid artery for a revision of hepatic arterial reconstruction after thrombosis in living related liver transplantation. PMID- 7482754 TI - Successful transplantation of a liver graft with a calcified hydatid cyst after back-table resection. PMID- 7482755 TI - Effect of calcium channel blockers on graft outcome in cyclosporine-treated renal allograft recipients. PMID- 7482756 TI - Immunosuppressive effects of leflunomide in experimental chronic vascular rejection. PMID- 7482757 TI - The ataxia-telangiectasia gene: a link between checkpoint controls, neurodegeneration and cancer. PMID- 7482758 TI - FISH analysis of interphase nuclei extracted from paraffin-embedded tissue. PMID- 7482759 TI - Combined isolation of nucleic acids and protein from small amounts of tissue. PMID- 7482760 TI - Human beta-actin retropseudogenes interfere with RT-PCR. PMID- 7482761 TI - A modified colony-filter-hybridization protocol for filamentous fungi. PMID- 7482762 TI - Gene number, methylation and biological complexity. PMID- 7482763 TI - Sex differentiation differs down under. PMID- 7482764 TI - What to do at an end: DNA double-strand-break repair. AB - Repairing chromosome breaks is essential to cell survival. A major lethal effect of ionizing radiation (IR) damage is the creation of double-strand DNA breaks. Recently, a number of mammalian cell mutants that are sensitive to IR damage have been described, revealing a unique repair pathway. The DNA-dependent protein kinase (DNA-PK) is necessary for double-strand-break repair and lymphoid V(D)J recombination. DNA-PK consists of three subunits: the Ku autoantigen heterodimer and a kinase (DNA-PKCS) that is deficient in mouse scid mutant cells. PMID- 7482765 TI - Genetic and environmental control of flowering time in Arabidopsis. AB - Plants detect fluctuations in environmental conditions and in response to these changes they promote or delay flowering. Some of the genes that are required to regulate flowering time in response to environmental changes were identified recently and cloned from Arabidopsis. The study of these genes is establishing connections between plant physiology and the expression of the homeotic genes that are required for floral development. PMID- 7482766 TI - Flagellar assembly in two hundred and fifty easy-to-follow steps. AB - The eukaryotic flagellum is a complex biochemical machine that moves cells or moves materials over the surface of cells, such as in the mammalian esophagus, oviduct or in protozoa. It is composed of over 250 polypeptides that must be assembled into a number of different structures and each structure must be attached with an exact periodicity along the microtubules. Once the flagellum is assembled, each of the components must act in concert and in three dimensions to produce a complex waveform. This review provides an outline of the composition and function of the different structures found in the flagella of Chlamydomonas. PMID- 7482767 TI - The function and evolution of Msx genes: pointers and paradoxes. AB - The Msx genes of vertebrates comprise a small family of chromosomally unlinked homeobox-containing genes related to the Drosophila gene muscle-segment homeobox (msh). Despite their ancient pedigree, the Msx genes are expressed in a range of vertebrate-specific tissues, including neural crest, cranial sensory placodes, bone and teeth. They are active in numerous systems, which have been used as models to study pattern formation and tissue interaction, and are, therefore, attracting a growing interest among developmental biologists. But beyond their presumed role as transcription factors, we do not know what their functions are in the cell or the embryo. Here, I review recent evidence that is beginning to address this problem and might eventually increase our understanding of how the vertebrate embryo has evolved. PMID- 7482768 TI - A tiger behind many doors: multiple genetic pathways to malignant glioma. AB - Malignant gliomas are the most common primary human brain tumours, but their classification remains controversial and effective therapies remain elusive. As a result, malignant gliomas have come under intense scientific scrutiny, in the hope of elucidating the molecular basis of glial tumourigenesis. These studies have yielded insights into the genetic events that underlie glioma formation and progression, and have shown multiple distinct genetic pathways that lead to the common malignant endpoint of glioblastoma multiforme. Such genetic pathways mirror clinicopathological avenues of glioma progression and suggest that molecular genetic approaches might have clinical utility in the coming years. PMID- 7482769 TI - A graphical interface for searching the human genome. PMID- 7482770 TI - Intercellular induction of homeotic gene expression in flower development. PMID- 7482771 TI - mRNA 3' ends in focus. PMID- 7482772 TI - On the nature of the mutation in the nude rat. PMID- 7482773 TI - A simple assay for puromycin N-acetyltransferase: selectable marker and reporter. PMID- 7482774 TI - Improved in vitro development of the chick embryo using roller-tube culture. PMID- 7482775 TI - Shoot development in plants: time for a change. AB - The shoot system in plants progresses through several discrete phases during its development. Changes in the timing of these phases have important consequences for the morphogenesis of the shoot and are likely to be important in plant evolution. Genetic analysis of phase change in herbaceous plants, such as maize and Arabidopsis, has defined some of the genes involved in this phenomenon and has suggested a model for the regulation of this key feature of plant development. PMID- 7482776 TI - Pax6: more than meets the eye. AB - The paired-box motif, originally defined in Drosophila segmentation genes is conserved in the Pax family of vertebrate developmental genes. Mutations that reduce Pax6 dosage cause dominantly inherited eye malformations in man and mouse. Remarkably, it has now been found that Drosophila has a homologue of Pax6, which also plays a key role in eye development. PMID- 7482777 TI - Induction and patterning of the vertebrate nervous system. AB - The formation of the vertebrate central nervous system depends on a series of inductive interactions. Ventral ectoderm gives rise to epidermis (skin), whereas dorsal ectoderm forms neurectoderm, which is regionalized along the anteroposterior axis in response to inductive signals from dorsal mesoderm. Specific neural cell types are established along the dorsoventral axis of the neural tube by inductive signals from the dorsal mesoderm and prospective epidermis. We review the induction and patterning of the nervous system, with an emphasis on molecules that might play a role in these interactions. PMID- 7482778 TI - Transcription: new insights from studies on Archaea. AB - Molecular-genetic analyses have revealed that the Archaea (archaebacteria) are phylogenetically distinct from both eukaryotes and eubacteria. Archaea lack nuclei and resemble eubacteria in morphology and genomic organization, but their molecular design shares many features with eukaryotes. Here, we review recent work that indicates that the archaeal transcriptional machinery is strikingly similar to the RNA polymerase I, II, and III systems of eukaryotic cell nuclei. These findings provide important insights into the evolution of the transcriptional apparatus. PMID- 7482780 TI - Online Mendelian Inheritance in Man. PMID- 7482779 TI - Dinucleotide relative abundance extremes: a genomic signature. AB - Early biochemical experiments established that the set of dinucleotide odds ratios or 'general design' is a remarkably stable property of the DNA of an organism, which is essentially the same in protein-coding DNA, bulk genomic DNA, and in different renaturation rate and density gradient fractions of genomic DNA in many organisms. Analysis of currently available genomic sequence data has extended these earlier results, showing that the general designs of disjoint samples of a genome are substantially more similar to each other than to those of sequences from other organisms and that closely related organisms have similar general designs. From this perspective, the set of dinucleotide odds ratio (relative abundance) values constitute a signature of each DNA genome, which can discriminate between sequences from different organisms. Dinucleotide-odds ratio values appear to reflect not only the chemistry of dinucleotide stacking energies and base-step conformational preferences, but also the species-specific properties of DNA modification, replication and repair mechanisms. PMID- 7482781 TI - Metamorphosis in Drosophila: from molecular biology to mutants. PMID- 7482782 TI - Centromere activation. PMID- 7482783 TI - Cloning wild-type chromosomal genes using transposon mutagenized DNA in gram negative bacteria. PMID- 7482784 TI - Rapid identification of ApoE alleles by multiple-single-strand conformation polymorphism (SSCP) analysis. PMID- 7482785 TI - Extracellular matrix and pattern in plant embryos: on the lookout for developmental information. AB - Cells of early plant embryos are totipotent yet follow different predetermined developmental pathways, producing embryos with well-defined autonomously developing tissues. Evidence is accumulating from a few model systems that information maintaining the determined state of a particular cell type is contained in a position-dependent manner in the extracellular matrix. This information can be released to signal back to the cell, thus controlling patterns of gene expression and influencing cell fate. PMID- 7482786 TI - The relationship between the flamenco gene and gypsy in Drosophila: how to tame a retrovirus. AB - For a long time, retroviruses have been considered to be restricted to vertebrates. However, the genome of insects contains elements like gypsy in Drosophila melanogaster that are strikingly similar to vertebrate proviruses of retroviruses, which were considered to be transposable elements. Recent results indicate that gypsy has infective properties and is therefore a retrovirus, the first to be identified in invertebrates. It is normally repressed by a host gene called flamenco, which apparently controls the transposition and infective properties of gypsy. This provides an exceptional experimental model to investigate the genetic relationships between retroviruses and their hosts. PMID- 7482787 TI - Chasing tails in ascidians: developmental insights into the origin and evolution of chordates. AB - The ascidian tadpole larva is regarded as a prototype of the ancestral chordate. Here we consider recent studies on the development of the tadpole larva that provide new insights into chordate origins and evolution. The notochord of ascidian larvae and vertebrates appear to be homologous structures based on their induction by endoderm and expression of the Brachyury (T) gene. The muscle cells of ascidian larvae also appear homologous to those of vertebrates based on their expression of bHLH myogenic and muscle-type actin genes, although they are specified by cytoplasmic determinants localized in the egg as well as embryonic induction. Studies of the tailless larvae of anural ascidians have resulted in the identification of Manx, a gene that may control tail development and evolution. These and other results support the ascidian tadpole prototype for the ancestral chordate. PMID- 7482788 TI - Mouse embryonic hematopoiesis. AB - The hematopoietic system of vertebrates is derived from the mesodermal germ layer in early embryogenesis. Various animal models have been used for the study of hematopoiesis, from early stages in the visceral yolk sac or its analog, to the later stages where hematopoiesis is observed in intraembryonic areas surrounding the aorta, genital ridge and pro/mesonephros. Using the mouse as a model, we describe what is known about mammalian embryonic hematopoiesis and put it in the context of hematopoietic cell formation in avian, amphibian and fish embryos. Evolutionary comparisons and recent experimental evidence show that there are two embryonic sites of developing hematopoietic activity in the mouse before fetal liver hematopoiesis and suggest that, during ontogeny, two successive waves of hematopoietic activity may contribute to the blood system of the adult. PMID- 7482789 TI - Human and rodent DNA sequence comparisons: a mosaic model of genomic evolution. AB - Three patterns of DNA sequence conservation have been identified from five human and rodent genomic sequence comparisons. First, a divergent pattern was observed in the noncoding sequences of the beta-globin and gamma-crystallin gene clusters, and second, a highly conserved pattern was observed in the noncoding regions of the T cell receptor C alpha-C delta, and the alpha- and beta-myosin-heavy-chain genes. A third, mixed pattern has also been found in the immunoglobulin IgH C mu C delta gene region. These three patterns of genomic evolution pose the fascinating possibility that large portions of the genome evolve at different rates. PMID- 7482790 TI - Comparative genomics, genome cross-referencing and XREFdb. PMID- 7482791 TI - Probing the function of novel genes in the nervous system: is antisense the answer? PMID- 7482792 TI - 'My most true mind thus makes mine eye untrue'. PMID- 7482793 TI - Retrograde signaling at GABAA-receptor synapses in the mammalian CNS. AB - Recent studies have emphasized the role of signals that travel from a target cell, in a retrograde direction, to cells that synapse on the target and influence their output. While the focus of most research on this topic has been on long-lasting alterations at excitatory synapses, evidence that implicates retrograde transmission in the transient reduction of GABAA-receptor-mediated inhibition in hippocampus and cerebellum has begun to accumulate. Brief depolarizations of the postsynaptic principal cells lead to increases in the intracellular concentration of Ca2+, and a reduction in GABAA-receptor-mediated responses for 1-2 min. No concomitant reduction in postsynaptic GABAA-receptor responsiveness has been detected. Rather, release of GABA from inhibitory interneuron terminals appears to be reduced. The properties of this 'depolarization-induced suppression of inhibition' might be appropriate for unique physiological roles. PMID- 7482794 TI - IP3 receptors and secretory granules. PMID- 7482795 TI - Glycation in neuropathies. PMID- 7482796 TI - Inositol monophosphatase--a putative target for Li+ in the treatment of bipolar disorder. AB - Attenuation of the phosphatidylinositol (PI) signal transduction pathway as a consequence of inhibition of inositol monophosphatase (IMPase) has been proposed as the mechanism for the efficacy of Li+ in the treatment of bipolar disorder. Nevertheless, Li+ also affects other aspects of PI signal transduction, and it is therefore not clear whether modulation of PI responses by Li+ can be attributed solely to inhibition of IMPase. However, inhibitors of IMPase mimic the effects of Li+ on some aspects of PI cell signalling, thus highlighting the potential of IMPase as a target for the treatment of bipolar disorder. The recent description of the three-dimensional structure of IMPase in conjunction with site-directed mutagenesis and kinetic studies has led to the elucidation of the enzyme mechanism. These structural and mechanistic data should prove useful in the development of novel inhibitors of IMPase that might ultimately prove useful clinically. PMID- 7482797 TI - A role for hippocampal opioids in long-term functional plasticity. AB - In the granule cells of the hippocampus, glutamate coexists with opioid peptides derived from the proenkephalin and prodynorphin genes. The functional significance of this coexistence has been unclear but recent evidence suggests that the dynorphins and enkephalins play a crucial role in regulating the efficiency of neurotransmission at granule-cell synapses. Together with evidence that the level of opioid activity in this pathway can change dramatically according to the physiological or pathological state of the tissue, this information focuses attention on granule-cell opioids as primary mediators of hippocampal plasticity. PMID- 7482798 TI - The Bcl-2 family of proteins, and the regulation of neuronal survival. AB - Bcl-2 is the founder member of a growing family of cytoplasmic proteins that modulate the responses of many cell types to the diverse extracellular signals that affect their survival. Although knowledge of the functions of these proteins has come largely from studying cells of the immune system, increasing evidence implicates these proteins in modulating neuronal survival. Several of these proteins are expressed in the nervous system, and experimental overexpression of Bcl-2 prevents the death of neurones deprived of particular neurotrophic factors in vitro, and rescues developing neurones that would otherwise die in vivo. PMID- 7482799 TI - Second-order neurones and receptor mechanisms in visual- and olfactory information processing. AB - The retina and olfactory bulb are relatively simple in their synaptic operation, and provide insight into the fundamental synaptic mechanisms of brain functions. In the visual system, bipolar cells receive glutamate input from photoreceptors, and segregate visual inputs into parallel ON and OFF responses to light exposure and termination. In the olfactory system, the mitral and tufted cells respond to excitatory inputs from olfactory receptor neurones, and undergo reciprocal regulation through dendrodendritic synapses with their associated granule cells. Recent studies of the synaptic operation and regulation of the bipolar, mitral and tufted cells, at the molecular level, have revealed the detailed synaptic mechanisms of the second-order neurones in the segregation and discrimination of sensory information, as well as the modulatory synaptic mechanism that is involved in olfactory-recognition memory formation. PMID- 7482800 TI - How multiple-synapse boutons could preserve input specificity during an interneuronal spread of LTP. AB - A model is proposed whereby the spread of long-term potentiation (LTP) between potentiated and neighboring neurons is initiated by a retrograde signal that is restricted to the synaptic clefts of the potentiated neurons. Next, a change, such as enhanced release of neurotransmitter, occurs in the presynaptic boutons that are associated with potentiated synapses. This change affects all synapses that are located on the potentiated boutons, and leads to LTP at synapses on neighboring neurons that share multiple-synapse boutons with the initially potentiated neurons. In this model, restricting the retrograde signal to the potentiated synaptic clefts ensures the axonal-input specificity of LTP, and the induction of the secondary LTP requires the same cellular mechanisms as those of induction of the primary LTP. PMID- 7482801 TI - The emergence and evolution of mammalian neocortex. AB - Cortical variation in mammals and other terrestrial vertebrates, re-examined by current comparative methodology (out-group analysis), indicates that separate lateral (olfactory), dorsal and medial (hippocampal) pallial or cortical formations arose with the origin of vertebrates. Although the exact origin of mammalian isocortex (so-called neocortex) is still disputed, it appears that the earliest mammals already had a six-layered isocortex with ten to 20 functional subdivisions. Among placental mammals, at least, isocortex has expanded numerous times, producing additional cortical subdivisions. Because these expansions were independent transformations of a simpler cortex, they produced subdivisions that are not homologous. PMID- 7482802 TI - Numbers, time and neocortical neuronogenesis: a general developmental and evolutionary model. AB - The number of neurons in the neocortex is the product of the size of the preneuronogenetic founder population, that is, the number of proliferative cells that are present at the onset of neuronogenesis, and neuronogenetic amplification occurring as neurons are being produced. The amount of neuronogenetic amplification is determined by changes in the output fraction, Q, from 0 to 1, over a fixed number of cell cycles. Greater neuronogenetic amplification would occur across species if the number of cell cycles during which Q < 0.5 increased. Since neither the length of the cell cycle nor the length of the neuronogenetic interval, that is, time per se, influence neuron number directly, it is speculated that changes in these parameters are essential to neuronal diversity. PMID- 7482803 TI - A small step for the cell, a giant leap for mankind: a hypothesis of neocortical expansion during evolution. AB - The more than 1000-fold increase in the cortical surface without a comparable increase in its thickness during mammalian evolution is explained in the context of the radial-unit hypothesis of cortical development. According to the proposed model, cortical expansion is the result of changes in proliferation kinetics that increase the number of radial columnar units without changing the number of neurons within each unit significantly. Thus, mutation of a regulatory gene(s) that controls the timing and ratio of symmetric and asymmetric modes of cell divisions in the proliferative zone, coupled with radial constraints in the distribution of migrating neurons, could create an expanded cortical plate with enhanced capacity for establishing new patterns of connectivity that are validated through natural selection. PMID- 7482804 TI - How do thalamic axons find their way to the cortex? AB - A cascade of simple mechanisms influences thalamic innervation of the neocortex. The cortex exerts a remote growth-promoting influence on thalamic axons when they start to grow out, becomes growth-permissive when the axons begin to invade, and later expresses a 'stop signal', causing termination in layer 4. However, any part of the thalamus will innervate any region of developing cortex in culture, and the precise topographic distribution of thalamic fibres in vivo is unlikely to depend exclusively on regional chemoaffinity. The 'handshake hypothesis' proposes that axons from the thalamus and from early-born cortical preplate cells meet and intermingle in the basal telencephalon, whereafter thalamic axons grow over the scaffold of preplate axons, and become 'captured' for a waiting period in the subplate layer below the corresponding part of the cortex. The bizarre pattern of development of thalamic innervation in the mutant reeler mouse provides strong evidence that thalamic axons are guided by preplate axons. PMID- 7482805 TI - Exuberant development of connections, and its possible permissive role in cortical evolution. AB - The callosal visual connections of the cat provide a model for studying the phenotypes of cortical axons and their differentiation. The terminal arbor of a callosal axon develops in several successive stages. At each stage, the arbor approximates the adult phenotype more closely. This is achieved through two mechanisms: (1) exuberant, but increasingly constrained, growth and (2) partial deletion of previously generated parts of the arbor. This differentiation is controlled by interactions of the axon with its cellular environment, and by visual experience. It might have played a permissive role in the evolution of the cerebral cortex by enabling adjustments of cortical connectivity to changes in the number, size, internal organization and cellular composition of cortical areas. PMID- 7482806 TI - The formation of a cortical somatotopic map. AB - The primary somatosensory cortex of small rodents is an isomorphic representation of the body surface. Similar representations are characteristic of the subcortical pathways, leading from the periphery to the cortex, and these representations develop in a sequence that begins at the periphery, and that ends in the cortex. Furthermore, central representations at all levels of the neural axis are altered by perinatal perturbations of the peripheral surface. This has led to the hypothesis that the periphery plays an instructional role in the formation of central neuronal structures. The morphology of this discrete organization has been examined thoroughly during the development of the thalamocortical projections. The mechanism(s) that underlies the formation of these representations remains unclear although some recent evidence suggests the involvement of activity-dependent processes that are modulated by 5-HT. PMID- 7482807 TI - The organization of neocortex in mammals: are species differences really so different? AB - By examining a variety of mammals, it is possible to determine common features of cortical organization, and from these infer homologies across species. Such analysis also enables differences in the organization of the neocortex to be identified. Species differ in the amount of cortex that is devoted to a particular sensory system, in the size and configuration of a cortical field, in the number of cortical fields, and in the pattern of connections of homologous fields. It is suggested that the plan of organization that is retained is the result of homologous developmental events, and that modifications to this plan are generated by a limited set of mechanisms. These types of changes to the common network might account for the sensory and behavioural diversity that is observed in extant mammals. PMID- 7482808 TI - Theory of mind: evolutionary history of a cognitive specialization. AB - Traditional analyses of the evolution of intelligence have emphasized commonality and continuity among species. However, recent research suggests that humans might have specialized in a particular kind of intelligence that is related to understanding mental states such as desires, intentions and beliefs. Data indicate that the ability to reflect on one's own mental states, as well as those of others, might be the result of evolutionary changes in the prefrontal cortex. Behavioral studies in children and chimpanzees reveal both similarities and striking differences in the developmental pathways that lead to theory-of-mind capacities. Humans and great apes share many ancient patterns of social behavior, but it is too early to be certain if they interpret them in the same manner. Humans might have evolved a cognitive specialization in theory of mind, forever altering their view of the social universe. PMID- 7482809 TI - Antiviral antibodies in transplantation. AB - Antiviral antibodies clearly play a role in organ transplantation. The only antibody with a licensed indication for this population is CMVIG; however, the use of HBIG has become the standard of care for those patients at risk for HBV recurrence, and the use of VZIG would appear to be indicated for exposures to varicella-zoster in patients lacking immunity. Future studies defining the role of antiviral antibodies for therapy are necessary, and the role of antibody to prevent the EBV PTLD remains to be explored. PMID- 7482810 TI - The potential role of therapeutic antibodies in the regulation of rejection. PMID- 7482811 TI - Genetically engineered proteins for immunoregulation. PMID- 7482812 TI - Prophylactic cytomegalovirus management strategies. PMID- 7482813 TI - Role of immunoglobulins and new antivirals in treatment of cytomegalovirus infection. PMID- 7482815 TI - Recurrent cytomegalovirus disease in solid-organ transplant recipients. AB - Recurrent CMV disease has been reported to occur in 6% to 59% of solid-organ transplant recipients. The consequences of recurrent CMV disease after the treatment of the first CMV disease episode in patient and graft survival have not been fully elucidated. Additionally, few epidemiologic data are available regarding predictors of recurrent CMV disease. In three of four studies that investigated predictors for recurrent CMV disease, primary CMV infection (D+/R-) was associated with a higher rate of recurrent CMV disease among liver, intestinal, and solid-organ transplant recipients in general. Antirejection therapy was associated with recurrent CMV disease in two of three studies that examined this variable as a possible predictor of recurrent CMV disease. Further studies are needed to define the subgroup of solid-organ transplant recipients at higher risk for recurrent CMV disease, for whom more intensive or novel prophylactic, therapeutic, or diagnostic cost-effective strategies are warranted. PMID- 7482814 TI - Cytomegalovirus prophylaxis with ganciclovir, acyclovir, and CMV hyperimmune globulin in liver transplant patients receiving OKT3 induction. PMID- 7482816 TI - Risk factors and pathogenesis of posttransplant lymphoproliferative disorders. PMID- 7482817 TI - Pretransplant assessment of the risk for posttransplant lymphoproliferative disorder. PMID- 7482819 TI - Increased late rejection in cytomegalovirus-immunoglobulin-M-positive heart transplantation patients. PMID- 7482818 TI - Effectiveness of combination prophylaxis with cytomegalovirus hyperimmune globulin and acyclovir in the high-risk kidney transplant recipient. PMID- 7482820 TI - Cytomegalovirus hyperimmune globulin for CMV disease refractory to ganciclovir in renal transplantation. PMID- 7482821 TI - Prevention and treatment of cytomegalovirus disease in thoracic organ transplant patients: evidence for a beneficial effect of hyperimmune globulin. PMID- 7482822 TI - Transmembrane signaling requirements of T cells: implications for regulation of alloimmunity. PMID- 7482823 TI - Successful treatment of Epstein-Barr virus infection with ganciclovir and cytomegalovirus hyperimmune globulin following kidney transplantation. PMID- 7482824 TI - Review and preview of anti-T-cell antibodies. PMID- 7482825 TI - The Banff classification of renal allograft pathology: where do we go from here? PMID- 7482826 TI - Transplantation of single pediatric kidneys into adult recipients--a 12-year experience. PMID- 7482827 TI - Kidney transplantation in patients older than 60 years of age--is it worth it? PMID- 7482828 TI - Mannitol infusion and decreased incidence of allograft acute renal failure. PMID- 7482829 TI - A report of 26 cases of renal transplantation in children. PMID- 7482831 TI - Triple therapy with cyclosporine A in kidney transplant patients. PMID- 7482830 TI - Quantitative determination of urine proteins: a rapid, noninvasive, sensitive, and inexpensive method to monitor renal grafts. PMID- 7482833 TI - The effect of DST in kidney transplantation between spouses. PMID- 7482832 TI - Factors affecting mortality in kidney transplantation. PMID- 7482834 TI - Serum erythropoietin levels after renal transplantation with various graft outcomes. PMID- 7482835 TI - The efficacy and safety of fluvastatin in hypercholesterolemia in renal transplant recipients. PMID- 7482836 TI - Musculoskeletal problems after kidney transplantation. PMID- 7482839 TI - Renal transplantation from living related and unrelated donors. PMID- 7482837 TI - Cryofiltration apheresis for treatment of cryoglobulin-induced glomerulonephritis in renal transplant. PMID- 7482838 TI - Pregnancy in renal transplantation recipients: outcome and complications in 44 pregnancies. PMID- 7482841 TI - Autotransfusions for surgical operations before renal transplantation. PMID- 7482840 TI - Epileptic seizure in chronic renal failure and transplanted patients. PMID- 7482843 TI - The effect of kidney donation on total renal function. PMID- 7482842 TI - Results of kidney transplantation from living unrelated donors. PMID- 7482844 TI - Renal transplantation using ureteral stents. PMID- 7482845 TI - Does kidney donation threaten the quality of life of the donor? PMID- 7482847 TI - Assessment of quality of life after kidney transplantation. PMID- 7482846 TI - The effect of preoperative structured teaching on renal transplantation outcome. PMID- 7482848 TI - Early experience of renal transplantation in hepatitis C patients. PMID- 7482849 TI - Is needle biopsy mandatory for donor selection in live donor segmental liver transplantation? PMID- 7482850 TI - Vascular complications following 200 liver transplants. PMID- 7482851 TI - The impact of hepatic function on infection following liver transplantation. PMID- 7482852 TI - Thermodiffusion as a novel method for continuous monitoring of the hepatic microcirculation after liver transplantation. PMID- 7482853 TI - Analysis of extravascular endothelin levels in UW solution and rinsing effluent of porcine liver grafts after cold storage. PMID- 7482854 TI - Liver transplantation in Turkey. PMID- 7482855 TI - Report of the first liver transplants in Iran. PMID- 7482856 TI - Intramyocardial electrograms for the monitoring of allograft rejection after heart transplantation using spontaneous and paced beats. PMID- 7482857 TI - Diltiazem prevents accelerated graft coronary artery disease in heart transplant recipients. PMID- 7482858 TI - Effect of advanced age on short- and long-term outcome of heart transplantation. PMID- 7482859 TI - Treatment of leukopenia with granulocyte-macrophage colony-stimulating factor after heart transplantation. PMID- 7482861 TI - Application of electron paramagnetic resonance for evaluation of acute rejection of the transplanted heart. PMID- 7482863 TI - Islet cell implantation into the pouched amniotic membrane on the side of the portal vein in dogs. PMID- 7482860 TI - Ex vivo pig to human heart transplantation: a histological study. PMID- 7482862 TI - Pancreatic transplantation. PMID- 7482864 TI - Autotransplantation of unpurified pancreatic islets of Langerhans into different sites in the canine model. PMID- 7482865 TI - Bone marrow transplantation. PMID- 7482866 TI - Prevention of graft-versus-host disease after bone marrow transplantation. PMID- 7482867 TI - Characterization of a murine monoclonal antibody reactive with a leukocyte common antigen and its application to bone marrow transplantation. PMID- 7482870 TI - Mechanism of enhancement in allografts: alloantibody does not accumulate in the graft but forms complexes with lymphocytes. PMID- 7482871 TI - Developments in immunosuppressive therapy. PMID- 7482869 TI - Pretransplant IgA-anti-hinge and IgA-anti-Fab autoantibody activity is associated with good kidney graft survival. PMID- 7482872 TI - Induction immunosuppression with antithymocyte globulin in renal transplantation using a variable dose according to the absolute number of CD3+ T cells. PMID- 7482868 TI - Bone marrow transplantation for thalassemia in Shiraz. PMID- 7482873 TI - Clinical experience in transferring kidney transplant patients from sandimmun to sandimmun neoral--results after 6 months. PMID- 7482874 TI - Modulation of cytotoxicity against donor HLA markers by posttransplant sera from renal allograft recipients. PMID- 7482875 TI - Retrospective study of cyclosporine-prednisolone versus cyclosporine-azathioprine prednisolone immunosuppression in living donor kidney transplantation. PMID- 7482877 TI - Conversion of cyclosporine to azathroprine in renal transplant recipients. PMID- 7482876 TI - Influence of HLA mismatching on the incidence of cytomegalovirus infection in living-related renal transplantations. PMID- 7482878 TI - Organ shortage and a long waiting list allow local kidney allocation to insure both short ischemia time and good HLA-A, B, DR matching. PMID- 7482879 TI - Microbiologic analysis of preservation solutions in cadaveric kidney transplantation. PMID- 7482882 TI - Infectious complications in the first 100 days after renal transplantation. PMID- 7482880 TI - Allopurinol inhibits endothelin release after mesenteric ischemia-reperfusion injury. PMID- 7482881 TI - Prevention of calcium influx does not inhibit endothelin release. PMID- 7482883 TI - Treatment of cytomegalovirus infections with gancyclovir during OKT3 administration. PMID- 7482884 TI - Ureteral complications of renal transplant surgery. PMID- 7482885 TI - Urinary tract infection vs renal allograft rejection. PMID- 7482886 TI - A case report of Kala-Azar in a kidney transplant patient. PMID- 7482887 TI - Flank versus transabdominal living donor nephrectomy: a randomized clinical trial. PMID- 7482888 TI - Autosomal dominant polycystic kidney disease-associated glomerulonephritides. PMID- 7482889 TI - Treatment of post-renal-transplant erythrosis: ACE inhibitors or theophylline? PMID- 7482890 TI - Cyclosporine-induced hyperuricemia and gout in renal transplants. PMID- 7482891 TI - Long-term nephrotoxicity of low doses of cyclosporine in heart transplant recipients. PMID- 7482892 TI - High incidence of posttransplant diabetes mellitus in renal transplant recipients on triple-drug immunosuppression. PMID- 7482894 TI - Cutaneous manifestations in renal transplant patients. PMID- 7482893 TI - Nocardiosis: report of four cases in renal transplant recipients. PMID- 7482895 TI - CNS infections in renal transplant recipients. PMID- 7482896 TI - Erythrocytosis in renal transplant recipients. PMID- 7482897 TI - The need to increase transplantation activity in developing countries. PMID- 7482898 TI - The use of plasmapheresis and systemic heparinization in treating glomerular microthrombi in renal transplants. PMID- 7482899 TI - Use of intraoperative ultrasound in a renal transplant recipient with renal artery stenosis. PMID- 7482900 TI - Detection of lipid peroxidation products by malondialdehyde (MDA-TBA reaction) in organ transplantation. PMID- 7482901 TI - Endothelin concentration in porcine hepatic tissue during ischemia and reperfusion. PMID- 7482902 TI - Induction of soft connective tissue formation by various prosthetic implants. PMID- 7482903 TI - Plasma membrane DNA in autoimmune diseases. PMID- 7482904 TI - Role of collagen in soft connective tissue wound healing. PMID- 7482905 TI - Kidney transplantation in Kermanshah, Iran: a 5-year experience. PMID- 7482907 TI - Cadaver kidney transplantation in Turkey. PMID- 7482906 TI - Experience with 300 renal transplants in Shiraz, Iran. PMID- 7482908 TI - First report of nine consecutive multiorgan cadaver donor transplants in Iran. PMID- 7482909 TI - Historical evolution of kidney and liver transplantation in Turkey. PMID- 7482910 TI - Living-related, cadaveric, and living unrelated donor kidney transplants: a comparison study at King Fahad Hospital, Riyadh. PMID- 7482912 TI - Renal transplantation in Pakistan. PMID- 7482911 TI - Immunosuppressive protocols at transplantation centers in Turkey. PMID- 7482914 TI - Is myocardial function influenced by neutrophil activation during ischemia and reperfusion in coronary artery surgery? PMID- 7482913 TI - Variations of glutathione and vitamin E concentrations after hypothermic storage in Euro-Collins solution and reperfusion of the rabbit kidney. PMID- 7482915 TI - Ischemia time dependency of reperfusion injury following complete arterial occlusion of the rat small intestine. PMID- 7482916 TI - Intermittent ischemia of the heart. PMID- 7482917 TI - Effect of warm ischemia on insulin secretion in non-heart-beating donors. PMID- 7482918 TI - Surgical management to avoid severe postreperfusion syndrome: controlled limb reperfusion. PMID- 7482920 TI - Pathophysiology of myocardial reperfusion injury: role of oxygen free radicals. PMID- 7482919 TI - Cytoprotective effects of a stable prostacyclin analog and a calcium channel blocker: a study on isolated rat hepatocytes. PMID- 7482921 TI - Element changes measured by X-ray microanalysis after cold ischemia and reperfusion: relation to recovery of function. PMID- 7482922 TI - Myocardial protection by nucleoside transport inhibition. PMID- 7482923 TI - Reperfusion syndrome in skeletal muscle after prolonged incomplete ischemia: reduction by controlled limb reperfusion. PMID- 7482924 TI - Controlling oxygen content during cardiopulmonary bypass to limit reperfusion/reoxygenation injury. PMID- 7482925 TI - Which cardioplegia for the hypertrophied myocardium? A prospective clinical study of four preservation techniques. PMID- 7482926 TI - Expression of intercellular adhesion molecule-1 and CD4/CD8 lymphocyte markers in perioperative liver allograft biopsies. PMID- 7482927 TI - Cytokine gene expression and eicosanoid production in renal reperfusion injury. PMID- 7482928 TI - Cardiac electrophysiologic and ultrastructural effects of platelet-activating factor and its antagonist BN 52021. PMID- 7482929 TI - Stimulation and modulation of mitochondrial radical generation during reperfusion injury of ischemic hearts. PMID- 7482930 TI - Fructose protects rat hepatocytes during hypoxia and improves protein synthesis recovery during reoxygenation. PMID- 7482931 TI - Potential role of cyclosporin A-sensitive mitochondrial channels in ischemia reperfusion injury. PMID- 7482932 TI - Mitochondrial oxidative phosphorylation injuries occurring in situ and in vitro. PMID- 7482933 TI - Mitochondrial function and cell injury in single cardiac myocytes exposed to anoxia and reoxygenation. PMID- 7482934 TI - Reperfusion injury in skeletal muscle. PMID- 7482935 TI - Damage to skeletal muscle induced by prolonged ischemia and reperfusion. PMID- 7482936 TI - Lazaroid U74389G attenuates skeletal muscle reperfusion injury in a canine model. PMID- 7482937 TI - Allopurinol improves postischemic skeletal muscle performance and endurance but not high-energy phosphate levels. PMID- 7482938 TI - Interaction between nitric oxide and superoxide: role in modulating leukocyte adhesion in the postischemic microvasculature. PMID- 7482939 TI - Platelet-activating factor antagonist TCV-309 protects rat kidneys against ischemia-reperfusion injury. PMID- 7482940 TI - Intravital microscope studies of the ischemically injured rat kidney during the early phase of reperfusion. PMID- 7482941 TI - Contrast-enhanced magnetic resonance imaging with intracellular contrast agents- a new way of monitoring hepatic reperfusion injury? PMID- 7482942 TI - Regulation of complement activation in rat liver ischemia and reperfusion: expression of endothelial CD59 (RIP). PMID- 7482943 TI - Role of xanthine oxidase activity in tissue damage of rat liver after ischemia. PMID- 7482944 TI - Metabolism of action of the prostacyclin analogue iloprost in hypothermic rat livers. PMID- 7482945 TI - Metabolic considerations of histidine-based solutions for liver storage. PMID- 7482946 TI - Protective effect of EGb 761 on ischemia-reperfusion damage in the rat retina. PMID- 7482947 TI - Microcirculatory disturbances secondary to ischemia-reperfusion. PMID- 7482948 TI - Intraoperative assessment of liver perfusion by laser Doppler imaging in an animal model. PMID- 7482949 TI - University of Wisconsin solution provides better lung preservation in human lung transplantation. PMID- 7482950 TI - L-arginine deficiency and hemodynamic changes as a result of arginase efflux following orthotopic liver transplantation. PMID- 7482951 TI - Cold and warm renal ischemia and postoperative survival in rabbits with autotransplanted kidneys. PMID- 7482952 TI - Protection from ischemia-reperfusion damage in patients with stroke: the role of rutin and GSH. PMID- 7482953 TI - Free oxygen species (FOS), FOS-scavenging enzyme P-selectin and monocyte activity in cell populations aspirated from early human renal allografts. PMID- 7482954 TI - Antioxidant defense and free radical production in a rabbit model of kidney ischemia-reperfusion. PMID- 7482956 TI - Categories of non-heart-beating donors. PMID- 7482955 TI - Beneficial effect of the platelet-activating factor antagonist WEB 2170 on reperfusion injury after orthotopic liver transplantation. PMID- 7482957 TI - Non-heart-beating donors: methods and techniques. AB - In light of the shortage of kidneys from HBDs the use of NHBDs can be considered essential. Emergency in situ preservation using a DBTL catheter offers the possibility of procuring kidneys from otherwise unsuitable donors. The procedure requires perfect organization and immediate availability of trained personnel and equipment. A standardized protocol is essential. The procedure is a valuable asset in emergency and transplant surgery. PMID- 7482958 TI - Experience with liver and kidney allografts from non-heart-beating donors. PMID- 7482959 TI - Organ procurement from non-heart-beating donors through in situ perfusion or total body cooling. PMID- 7482960 TI - Successful transplantation of kidneys harvested from cadaver donors at 71 to 259 minutes following cardiac arrest. PMID- 7482961 TI - Posttransplant long-term outcome of kidneys obtained from asystolic donors maintained under extracorporeal cardiopulmonary bypass. PMID- 7482962 TI - Viability assessment of non-heart-beating donor kidneys during machine preservation. PMID- 7482963 TI - Newly developed four-lumen catheter for in situ renal perfusion of non-heart beating donors that provides perfusion pressure monitoring. PMID- 7482964 TI - Time is of the essence: the pressing need for comprehensive non-heart-beating cadaveric donation policies. PMID- 7482965 TI - Non-heart-beating kidney donation in uncontrolled donor procedures. PMID- 7482966 TI - Ethical considerations in procuring organs from non-heart-beating donors after sudden cardiac death. PMID- 7482967 TI - The introduction of a non-heart-beating donation program and the medical ethics committee. PMID- 7482968 TI - Legal issues concerning the use of non-heart-beating donors. PMID- 7482969 TI - Legal aspects of non-heart-beating donors: the Madrid solution. PMID- 7482970 TI - Non-heart-beating donors: the Leicester experience. PMID- 7482971 TI - Transplantation of kidneys from non-heart-beating donors: an update. PMID- 7482972 TI - Retrospective analysis of the outcome of transplantation of non-heart-beating donor kidneys. PMID- 7482973 TI - Cadaveric kidney transplantation activities in Istanbul. PMID- 7482974 TI - Long-term follow up of transplanted non-heart-beating donor kidneys. PMID- 7482975 TI - Results of transplantation with non-heart-beating donors. PMID- 7482976 TI - Organ and tissue procurement rates improve after professional healthcare education by hospital development coordinators and time sensitive requesting. PMID- 7482977 TI - Cost analysis of transplantation with ischemically damaged kidneys: preliminary results. PMID- 7482978 TI - Immediate function and cost comparison between ice storage and pulsatile preservation in kidney recipients at one hospital. PMID- 7482979 TI - Statement on non-heart-beating donor programs. PMID- 7482981 TI - Inverse agonism: theory and practice. PMID- 7482980 TI - Cloning of human prostanoid receptors. PMID- 7482982 TI - Pharmacological proteus? PMID- 7482983 TI - Endogenous ligands and inverse agonism. PMID- 7482984 TI - Molecular mechanisms of agonism. PMID- 7482985 TI - Benefits of ethological analysis of behaviour. PMID- 7482986 TI - Use of an antagonist for estimating the degree of agonist stimulation during physiological release. AB - Antagonist activity can be measured either as the percentage reduction in the effect of an agonist or as a dose ratio. In this article, Dick Barlow explains how these are connected and how the relationship between the concentration of antagonist producing 50% inhibition (IC50) and the antagonist equilibrium constant (Ki) involves the degree of agonist stimulation ([A]/[A50]), which is the concentration of agonist expressed as a multiple of the concentration producing a half-maximal response. If Ki is known it is possible to use the IC50 value of the antagonist to estimate the degree of agonist stimulation in physiological experiments where there is neuronal or hormonal release of the agonist. PMID- 7482987 TI - The role of vanadium in the management of diabetes. AB - Diabetes mellitus results from an absolute or relative deficiency in insulin secretion and a resistance of target tissues to the action of insulin, in proportions that vary with the type of the disease. The shortage of insulin can be corrected by administration of exogenous insulin or stimulation of pancreatic beta-cells with sulphonylureas. However, insulin resistance remains a major therapeutic problem. Here, Sonia Brichard and Jean-Claude Henquin review the recent discoveries that indicate a possible role for vanadium in management of the disease. In vitro, vanadium salts mimic most effects of insulin on the main target tissues of the hormone, and in vivo they induce a sustained fall in blood glucose levels in insulin-deficient diabetic rats, and improve glucose homeostasis in obese, insulin-resistant diabetic rodents. Recent short-term clinical trials with vanadium salts also seem promising in type II (non-insulin dependent) diabetic patients in whom liver and peripheral insulin resistance was attenuated, indicating the therapeutic potential of vanadium salts, pending demonstration of their long-term innocuity. PMID- 7482988 TI - Novel dopamine receptors half a decade later. AB - Only half a decade ago, the effects of dopamine were all attributed to activation of two receptor subtypes, the D1 and D2, with opposing effects on adenylate cyclase, and for which apparently selective ligands were available. From the end of 1988, however, the application of homology cloning techniques starting from sequences of the seven transmembrane domain catecholamine receptors, particularly that of the D2 receptor, led to the identification of 'novel', previously uncharacterized dopamine receptors. In this article, Pierre Sokoloff and Jean Charles Schwartz discuss the functional significance of such diversity, as well as the new therapeutic perspectives it offers. PMID- 7482989 TI - Fishing for drugs from the sea: status and strategies. AB - Marine organisms represent an enormous, essentially unexploited, resource of natural products. Globally, the race to develop marine-derived drugs is well under way with many pharmaceutical companies positioning themselves to reap large profits by the exploitation of the ocean's rich chemical diversity. Targeted strategies, often in combination with high-throughput screening, are being employed in this hunt for novel pharmacotherapeutic agents. David de Vries and Phil Beart examine the potential, problems and technologies of an international pharmaceutical search that has important ethical considerations. PMID- 7482990 TI - Adenosine, antioxidant enzymes and cytoprotection. PMID- 7482991 TI - 3D-QSAR: a current perspective. PMID- 7482992 TI - Selectivity of the neuroprotective agent lifarizine. PMID- 7482993 TI - Nomenclature of nitrogen oxide-containing molecules. PMID- 7482994 TI - Neurosteroids and GABAA receptor function. AB - In 1984, a potent and selective interaction of the steroidal anaesthetic alphaxalone with the GABAA receptor was demonstrated. Subsequent studies established that certain naturally occurring steroids were potent positive allosteric modulators of the GABAA receptor. Although peripheral endocrine glands are an important endogenous source, the brain can synthesize 'neurosteroids', and these have the potential to influence the activity of the GABAA receptor in the CNS. Systemic administration of steroids have clear behavioural effects. In this article, Jeremy Lambert and colleagues review recent advances in this field and discuss the therapeutic potential of this novel, non-genomic effect of steroids and investigate whether they may influence behaviour under physiological, or pathophysiological, conditions. PMID- 7482995 TI - Cysteinyl leukotrienes in asthma: old mediators up to new tricks. AB - The cysteinyl leukotrienes have long been suspected to play a role in the pathogenesis of asthma. This speculation was based largely on their release in human lung following antigen challenge as well as their potent bronchoconstrictor activity. However, there is increasing evidence that the cysteinyl leukotrienes also produce several pro-inflammatory effects and alter the activity of neuronal pathways in the airways. Douglas Hay, Theodore Torphy and Bradley Undem review these recent data and discuss the therapeutic possibilities of cysteinyl leukotriene receptor antagonists and 5-lipoxygenase inhibitors. PMID- 7482997 TI - Implications of prolonged expression of Fos-related antigens. AB - The AP-1 transcription factors are composed of the Fos and Fos-related antigens as well as Jun and related proteins. These factors have been extensively studied in many diverse paradigms using acute stimuli. Recent attention has focussed on long-term elevation of Fos-related antigens in the CNS, and this is discussed by Keith Pennypacker, Jau-S. Hong and Michael McMillian. Repeated or chronic treatment elevates Fos-related antigen levels for days in many different brain regions. Both direct and indirect stimulation are responsible for the protracted increase in Fos-related antigen-immunoreactive proteins, which may modulate late onset genes involved in neuroplasticity. Understanding the role of these factors in long-lasting or permanent disease states may provide insight into potential therapeutic strategies to treat chronic CNS disorders. PMID- 7482996 TI - Na+ channels as targets for neuroprotective drugs. AB - Drugs that block voltage-dependent Na+ channels are well known as local anaesthetics, antiarrhythmics and anticonvulsants. Recent studies show that these compounds also provide a powerful mechanism of cytoprotection in animal models of cerebral ischaemia, hypoxia or head trauma. In this article Charles Taylor and Brian Meldrum review evidence indicating that Na+ channel modulators are neuroprotective and describe recent ideas for the molecular sites of action of voltage-dependent Na+ channel blockers. Clinical trials with several compounds are now in progress for stroke and traumatic head injury, and the therapeutic potential for this group of compounds is discussed. PMID- 7482998 TI - Comparison of three malaria chemoprophylactic regimens in residents in east, central and southern Africa. A prospective, randomized multicentre trial in The Netherlands. AB - There is much confusion about the most effective malaria chemoprophylactic regimen for travellers to chloroquine-resistant areas. For residents, the problem is even more confused. A prospective, multicentre trial was performed between 1987 and 1989 to assess the efficacy of three different malaria chemoprophylactic regimens (chloroquine 300 mg weekly combined with proguanil 100 mg daily, chloroquine 300 mg weekly combined with proguanil 200 mg daily and proguanil 200 mg daily only) in Dutch expatriates, who were departing for a stay of more than one year in East, Central or Southern Africa. Prophylaxis failures (defined as Plasmodium falciparum present in the blood film) were distinguished from failures of compliance by measuring whole-blood drug levels, taken at the same time as the blood slide and sent as filterpaper blood spots. The data of 200 expatriates could be analysed; the overall response rate was 52%. Twenty-six (13%) suffered from a fever; in two of them the blood slide contained P. falciparum. One subject took chloroquine 300 mg weekly with proguanil 200 mg daily, the other 200 mg proguanil daily. Assessment of compliance was possible in 10 of the 26 subjects with a fever; five (50%) were below and five (50%) were above the limit of 0.19 mumol/l. Due to the low incidence of prophylaxis failures, calculation of risks is unreliable. There were strong indications that compliance decreased with time. The ultimate cooperation needed for confirmation of prophylaxis failures and breakthroughs failed. Several factors which could have contributed to this lack of cooperation are discussed. PMID- 7482999 TI - Severe malaria in children at Port Moresby General Hospital, Papua New Guinea. AB - The demographic and clinical features of severe malaria in children on the south coast of Papua New Guinea have never been clearly documented. This prospective study sought to define the associations between ethnic origin, domain, age, nutritional status and severe malaria in this group and to assess significant clinical features, evaluate the use of a coma score as a prognostic indicator in cerebral malaria and to determine the ultimate outcome. Twenty patients with severe malaria (17 cerebral malaria and 3 severe anaemia) were studied. Their mean age of 4.96 years was significantly greater than that of matched controls with uncomplicated. Plasmodium falciparum infection with mean age 3.79 years (0.02 < p < 0.05). Nutritional status was not a significant independent risk factor when controlled against inpatients with other diagnoses. Low coma scores (Adelaide scale 4/14 or less) sensitively predicted the risk of dying vs survival. The mortality of 18% was comparable with other series. Current standard treatment with quinine and Fansidar was effective and no early recrudescence was encountered in the survivors. The degree of intermarriage and migration between regions precluded firm conclusions from being drawn as to the relevance of ethnic and geographical factors in the epidemiology of severe malaria in this region. PMID- 7483002 TI - Health problems of refugees in The Netherlands. AB - Research was carried out on the medical, i.e. somatic and mental, and social complaints of refugees in The Netherlands. This research consisted of a literature study and a retrospective and prospective, or cross-sectional patient study. The most important assumption which formed the base of the study was: refugees who underwent torture present the same medical and social complaints as refugees who were not tortured but underwent other forms of organized violence. For the cross-sectional study 156 refugees from the Middle East were interviewed. Not only was investigated whether the nature and extent of organized violence influenced the presentation of the refugees' medical and social complaints, but also whether other characteristics such as legal status and length of stay in The Netherlands were related with the refugees' medical and social complaints. The same data were sought for in literature. The results confirm the main assumption, as well as show that factors other than the traumatic experiences in the country of origin are related with the health problems of refugees. Although many studies in literature demonstrated that a relatively high percentage of refugees present a post traumatic stress disorder (DSM-III-R) these findings could not be confirmed. It is concluded that refugees undergo a sequence of traumatic experiences and stress before, during and after their flight into exile. Various pre- and post-migration factors are responsible for the presentation of aspecific physical and mental complaints and social problems of refugees. PMID- 7483000 TI - Splenomegaly in Baringo District, Kenya, an area endemic for visceral leishmaniasis and malaria. AB - The relationship between splenomegaly and visceral leishmaniasis (VL) was investigated during a cross-sectional study in 2,941 individuals in Baringo District, Kenya, where both malaria and VL are endemic. Spleen size was correlated with presence of malaria parasites in thick blood films and with evidence of present or past Leishmania donovani infection as determined by serology and history. Marked splenomegaly (Hackett grade 3 or greater) significantly correlated with present or previous leishmanial infection (chi 2 = 53.5; p < 0.001) whereas moderate splenomegaly (Hackett grade 1 or 2) significantly correlated with malaria parasitaemia (chi 2 = 73.03; p < 0.001). The presence of antimalarial antibodies did not contribute to the differentiation of the cause of splenomegaly. The diagnostic significance of splenomegaly in this population is discussed. PMID- 7483001 TI - The aetiologies of epilepsy in tropical Africa. AB - The prevention of epilepsy in tropical Africa is highly desirable because of the morbidity, mortality and social ostracisation that is associated with the disease. Such prevention depends on the identification of the aetiologies of epilepsy endemic to the region. There is a need for prospective epidemiological research to elucidate further the role of filariasis, cysticercosis, cerebral malaria and trypanosomiasis in the aetiology of epilepsy in tropical Africa. PMID- 7483003 TI - Pathways to psychiatric care in Ibadan, Nigeria. AB - Pathways patients take to psychiatric care will reflect the nature of the services available and the popular beliefs about mental illness. Studying the pathways may help in the identification of sources of delay in the receipt of care and suggest possible improvements. By studying the pathways of 159 patients to a tertiary psychiatric service in Nigeria, we show that traditional and religious healers are consulted at some stage by many patients with mental illness. Such healers are the first carers to a large proportion of the patients. These patients are not different from those who consult orthodox medical practitioners either in demographic features, presenting complaints, or nearness to service. Patients who consult traditional healers first tended to arrive at a tertiary psychiatric service much later than those who consult other carers. Our observations suggest that attempts to incorporate traditional medical care into the health care system must seek to improve their referral skill. PMID- 7483004 TI - Comparing liquid crystal thermometer readings and mercury thermometer readings of infants and children in a traditional African setting. Implications for community based health. AB - Liquid crystal thermometer (LCT) readings of skin temperatures were compared with mercury thermometer (MT) rectal temperature readings to assess the reliability of LCTs. Temperatures of 498 children were measured at two points in time. LCT skin temperature readings of children 0 to 52 months were on average 0.50 degrees C and 1.97 degrees C lower than MT rectal temperature readings. A strong correlation between temperature differences and LCT readings indicated that the greatest differences occurred at the lower LCT readings. These conclusions indicate LCT skin readings undermeasure temperature. Some of these differences were due to MTs not measuring temperatures below 35 degrees C. Children under 1 year of age had significantly greater differences than any other age group. Their LCT readings were, on average, 1.65 degrees C lower than their MT readings. Using MTs as a standard, LCTs were 100% sensitive and 92% specific for detecting children with hypothermia. LCTs were 38.5% sensitive and 100% specific for detecting fevers. These results suggest that LCTs leave undetected a large proportion of children who have fevers. However, they are sensitive for identifying children with hypothermia. A knowledge, attitude and practice (KAP) study indicated that local mothers can be identified who understand principles and procedures of LCTs, and accept them for health care of their child. PMID- 7483006 TI - Angiostrongylus costaricensis in a black-eared marmoset. AB - A case of Angiostrongylus costaricensis in a captive C. penicillata in a German zoological garden is described. Clinical symptoms were lacking, the morphological lesions consisted of intestinal granulomas harbouring rhabdidiform larvae. The adult worms were present in the ileo-caecal branches of the mesenteric artery. PMID- 7483008 TI - Unusual complications in benign tertian malaria. AB - An unusual case of malaria with Plasmodium vivax is reported which had complications classically seen with Plasmodium falciparum malaria. The complications were cerebral malaria, disseminated intravascular coagulation and adult respiratory distress syndrome. PMID- 7483005 TI - Tumour necrosis factor alpha in uncomplicated malaria in young adults. AB - The purpose of this study was: 1) to measure tumour necrosis factor alpha (TNF) in the plasma of Plasmodium falciparum infected subjects; and 2) to correlate the presence of TNF to symptomatology. Plasma from 77 malaria infected individuals (with malaria parasites) were assayed for TNF by ELISA. The mean age of the subjects under study was 16.36 +/- 0.80 (mean +/- SEM) years. Thirty-nine (51%) subjects had measurable plasma TNF. Taking symptomatology into account, 10 (59%) of the 17 asymptomatics and 29 (48%) of the 60 symptomatics had measurable plasma TNF. A risk ratio of 0.9 was obtained for the association between the detection of plasma TNF and the presence of symptoms. In plasma from 13 healthy controls no TNF was detected. The results suggest that if TNF plays a negative role in the pathogenesis of malaria, it must be in the presence of other predisposing factors. PMID- 7483007 TI - Fulminating strongyloidiasis complicating Indian kala-azar. AB - Infection with Strongyloides stercoralis in an immunocompetent host usually remains unrecognized. Sometimes, however, it may cause serious problems in the immunocompromised host due to its enhanced invasiveness. We present here details of three kala-azar patients with fulminating strongyloidiasis manifested by acute diarrhoea with blood and mucus, severe dehydration, cough and dyspnoea with low peripheral blood eosinophils and filariform larvae of S. stercoralis in faeces. No larvae could be detected in the sputum in spite of radiological evidence of infiltration. The impact of albendazole was dramatic with a disappearance of parasites from the faeces and of pulmonary infiltration and an increase in blood eosinophil count. It is emphasized that S. stercoralis infection in kala-azar patients should be considered a serious infection and treated accordingly. PMID- 7483011 TI - Usefulness of electron microscopy in the diagnosis of "small" round cell tumors of the sinonasal region. AB - The sinonasal region is known to harbor several types of tumors that belong to the general category of "small" round cell tumors and offer considerable diagnostic challenges. This study evaluated 33 cases of such tumors by electron microscopy to characterize their ultrastructural features in conjunction with immunohistochemistry, in an attempt to define diagnostic criteria of various types. Electron microscopy was useful in the proper classification of tumors in 27 cases: esthesioneuroblastoma (EN), 12; undifferentiated carcinoma, 6; melanoma, 3; lymphoma, 3; melanotic neuroectodermal tumor, 1; rhabdomyosarcoma, 1; and pituitary adenoma, 1. In the remaining six cases, the ultrastructural features were those of poorly differentiated carcinomas. They usually exhibited some epithelial characteristics as well as neuroendocrine features by immunohistochemistry and electron microscopy. These tumors could be best described as poorly differentiated neuro-endocrine carcinomas (malignant neuroepitheliomas). The most controversial diagnostic problems existed between the tumors categorized as esthesioneuroblastomas and neuroendocrine (NE) carcinomas. Esthesioneuroblastomas were characterized by uniform round nucleated cells with variable amounts of dendritic processes containing numerous dense core granules ranging from 150 to 350 nm in the perikarya and dendritic processes. Dendritic processes contained longitudinally arranged neural tubules and revealed an occasional synaptic junction. In three of the 12 cases of EN, cells with the appearance of sustentacular cells were recognized by electron microscopy. The NE carcinomas usually consisted of closely packed round cells with scanty cytoplasm that lacked any feature of neuroblastic cells. The tumor cells in this category often were epithelioid in appearance and exhibited a varying degree of cytokeratin positivity. Neuron-specific enolase was also positive in all cases, further suggesting their neuroepithelial nature. The greatest difference between EN and NE carcinomas was the absence of sustentacular cells in NE carcinomas. Immunohistochemical and electron microscopic studies are essential in the differential diagnosis of EN and NE carcinomas, because their microscopic appearance is very similar. The study indicates that EM is useful in the diagnostic categorization of sinonasal tumors of uncertain nature, particularly when it is used in conjunction with immunohistochemistry. PMID- 7483009 TI - Falciparum malaria, imported into The Netherlands, 1979-1988. II. Clinical features. AB - To study the clinical features in patients with falciparum malaria and the influence of chloroquine chemoprophylaxis on these features, a prospective study was carried out of all non-immune patients with falciparum malaria between 1979 and 1988. Three hundred and sixty-one consecutive non-immune patients with falciparum malaria who were seen at the outpatient-department for Tropical Diseases, Royal Tropical Institute, and Division of Infectious Diseases and Tropical Medicine of the University Hospitals of the University of Amsterdam, The Netherlands. Compliance with the recommended malaria prophylaxis was claimed by 47% (168/361); 24% (86/361) had not taken any chemoprophylaxis. The first group had a milder illness, less often parasitaemia > or = 1% and suffered less from complications. Comparison of the clinical features in patients who did not take prophylaxis and those who, during recommended prophylaxis, appeared to be infected with a chloroquine-resistant strain strengthened the evidence of a protective effect of chloroquine. Complications occurred in 14% of patients; pregnant women were at higher risk of complicated malaria; the case-fatality rate was low (0.3%). It is concluded that chloroquine prophylaxis still may have- depending on the degree of chloroquine-resistance--a protective effect on the clinical features and this modulating effect needs to be further investigated. PMID- 7483012 TI - Polymorphous low-grade adenocarcinoma arising in the nasal cavities with an associated undifferentiated carcinoma. AB - A case of polymorphous low-grade adenocarcinoma arising in the right nasal cavity of a male patient with previous history of kidney transplantation is reported. The tumor contained extensive areas of undifferentiated carcinoma, probably arising from the low-grade component. This combination of findings has not been previously reported in this variety of salivary gland tumor. PMID- 7483013 TI - Ultrastructural distinction of basaloid-squamous carcinoma and adenoid cystic carcinoma. AB - Basaloid-squamous carcinoma is a unique and infrequent variant of squamous cell carcinoma with a predilection for occurring in the upper aerodigestive tract. The cardinal diagnostic criteria are the presence of basaloid cells arranged in lobules or cords and a squamous component. Another feature that distinguishes basaloid-squamous carcinoma from conventional squamous cell carcinoma is the development of multiple foci of amorphous, eosinophilic, or mucinous materials among the basaloid tumor cells. It is this feature in basaloid-squamous carcinoma that causes confusion with adenoid cystic carcinoma, especially the solid variant of adenoid cystic carcinoma. To clarify the distinctive features of these two types of tumors, the ultrastructural findings in three basaloid-squamous carcinomas from the head and neck region were compared with three adenoid cystic carcinomas, one from the head and neck region and one each from trachea and bronchus. Widened intercellular spaces, tonofilaments, and replicated basal lamina were present in both types of tumor. However, the ultrastructural features of glandular differentiation were present only in adenoid cystic carcinoma, whereas the ultrastructural features of squamous cell differentiation were present only in basaloid-squamous carcinoma. The distinction between basaloid squamous carcinoma and adenoid cystic carcinoma is important since basaloid squamous carcinoma is a clinically aggressive, high-grade form of squamous cell carcinoma, generally with a worse prognosis than adenoid cystic carcinoma. PMID- 7483010 TI - Myoepithelioma: definitions and diagnostic criteria. AB - Due to their infrequency and multiplicity of histopathology, myoepitheliomas present difficulties in diagnosis and classification. Cellular varieties can be misdiagnosed as malignancies. Improvements in and clarification of diagnostic criteria are, therefore, required. A key to determining diagnostic criteria for myoepitheliomas is to study cellular morphology, cytoplasmic filament expression, and ultrastructural features of the nonluminal, i.e., neoplastic myoepithelial/basal, tumor cells of pleomorphic adenomas, and apply this information to defining myoepitheliomas. Cytologic and growth patterns of nonluminal cells in pleomorphic adenomas, including plasma-cytoid cells, are reflected in myoepitheliomas. Results also indicate that muscle-specific actin and myofilaments are expressed only in a proportion of cases, and generally in not more than 60-70% of nonluminal cells in pleomorphic adenoma; this also applies to benign and malignant myoepitheliomas. The absence of these markers does not exclude a diagnosis of myoepithelioma. Vimentin and glial acidic fibrillary protein, however, are strongly and diffusely expressed in the majority of pleomorphic adenomas and myoepitheliomas and are more reliable markers for these tumors than muscle-specific actin. Like so many other salivary gland tumors, myoepitheliomas present an equally complex histomorphology and variable expression of antigenic markers, only some of which are associated with myoepithelial and basal cells of the acini and ducts of the normal salivary gland. PMID- 7483014 TI - Recurrent cervical lymphadenopathy due to Ki-1 lymphoma in an elderly female. AB - A 70-year-old female presented with recurring cervical lymphadenopathy over a 5 year period. Intermittently, symptoms of respiratory and GI tract disease occurred, raising the possibility of malignancy. Investigation of these systems and of the breasts were negative. A lymph node biopsy was eventually done and showed intrasinusoidal proliferations of CD30-positive malignant cells which were also characterized ultrastructurally. Therapy with CHOP has produced a good initial response. The value of electron microscopy in diagnostic pathology is illustrated. PMID- 7483015 TI - Pleural mesothelioma with initial presentation as cervical lymphadenopathy. AB - A 41-year-old woman presented with a left neck mass. Light microscopy suggested metastatic large cell carcinoma, but subsequent electron microscopy demonstrated typical epithelial mesothelioma. Initial clinical investigations were negative and the primary pleural tumor remained inapparent for a further 8 months. The woman ultimately died from the effects of superior vena caval obstruction. Mesotheliomas frequently metastasize late in the course of the disease, but it is exceptional for them to present as a metastatic tumor of unknown primary origin. PMID- 7483016 TI - Malignant melanoma of soft parts involving the head and neck region: review of literature and case report. AB - Malignant melanoma of soft parts (MMSP) was originally described as a distinct entity by Enzinger in 1965 and was termed "clear cell sarcoma of tendons and aponeuroses" because of its association with tenosynovial structures. It has been shown immunophenotypically and ultrastructurally that this tumor is derived from neuroectoderm and shares a number of features with cutaneous melanoma. Over 95% of MMSPs present in the extremities, with the head and neck region (1.9%) being an unusual site. This study presents an additional case of MMSP of the head and neck region involving the posterior cervical region in a 15-year-old Hispanic male and reviews the literature on MMSP. Ultrastructural examination showed rudimentary cell attachments, smooth cell membranes, discontinuous basal lamina, scanty glycogen, and occasional premelanosomes in some tumor cells. Cytogenetic analysis showed a reciprocal translocation between the long arms of chromosomes 12 and 22 [t(12:22)(q13;q12.2)], characteristic for MMSP and not seen in cutaneous melanoma. Survival in MMSP has been correlated with tumor size, tumor necrosis, and ploidy status. Overall reported clinical outcome for this tumor is as follows: died of disease, 45%; alive with disease, 23%; no evidence of disease, 30%; and died of other causes, 2%. MMSP represents a distinct entity with a characteristic ultrastructural appearance and a tumor defining cytogenetic translocation. PMID- 7483017 TI - Signet-ring stromal and related tumors of the ovary. AB - The signet-ring stromal tumor of the ovary, described only twice previously, is an enigmatic lesion in respect to its cell lineage and the nature of the cytoplasmic vacuoles that cause the signet-ring cell appearance. The vacuoles were shown in the initial report to contain no lipid, mucoprotein, or glycogen, and the ovarian stromal cell was assumed to be the cell of origin. The present study describes three ovarian neoplasms in which the immunohistochemical and ultrastructural findings support a stromal origin for the cells, and a fourth tumor that simulated the other three closely on light microscopic examination but had several features supporting a sex-cord rather than stromal origin: immunohistochemical staining for keratin and alpha smooth muscle actin, and globular keratin staining, similar to that seen in granulosa cell tumors; and numerous prominent junctions including desmosomes on electron microscopy. In only one of the four tumors were the vacuoles similar ultrastructurally to those in the first reported case, appearing to result from generalized edema of the cytoplasmic matrix. In a second tumor, the vacuoles resulted from hydropic swelling of numerous mitochondria, which filled the cytoplasm of the cells. In the other two tumors, the vacuoles were not cytoplasmic, but cytoplasmic pseudoinclusions of edematous extracellular matrix. An additional interesting finding in one of the four tumors was the presence of hyaline globules within and adjacent to the signet-ring cells. Ultrastructural examination revealed the globules to be degenerating erythrocytes, many of which had been phagocytosed by the tumor cells. PMID- 7483018 TI - Endometrioid-like variant of ovarian yolk sac tumor with foci of carcinoid: an ultrastructural study. AB - The endometrioid-like variant of ovarian yolk sac tumor is a rare neoplasm composed partially or completely of glandular tissue resembling endometrioid carcinoma, but identifiable histologically and immunohistochemically as an extraembryonic endodermally differentiated germ cell tumor. Only nine examples of this neoplasm have been published previously, and none was studied by electron microscopy. One of these rare neoplasms, the first documented to have foci of carcinoid, has recently been observed and studied ultrastructurally. Electron microscopic observations showed that the main component of the neoplasm had glandular epithelial cells with nuclear and cytoplasmic features difficult to distinguish from the cells of Mullerian endometrioid and endometrial adenocarcinoma, but a minor component of mucinous adenocarcinoma was confirmed as intestinal by the epithelial cells having characteristic microvilli with filamentous cores and rootlets. In addition, neuroendocrine cells present in the glands and in nests were readily identified ultrastructurally by their dense-core granules. Electron microscopy may be useful in supplementing light microscopic and immunohistochemical examination of this variant of yolk sac tumor. PMID- 7483019 TI - Monomorphic (basal cell) salivary adenoma of ovary: report of a case. AB - A pregnant 26-year-old woman presented with an asymptomatic 10-cm left ovarian mass diagnosed at 19 weeks gestation. On removal, the tumor was shown to be a cribriform variant of basal cell salivary adenoma with no other teratomatous or germ cell tumor elements present. The pregnancy continued uneventfully to term and the patient remains symptom-free 10 months after diagnosis. Tissues of salivary type are occasionally observed within mature cystic teratomas of the ovaries and are presumed to have given rise to the present lesion, which is thought to be the first reported case of a neoplasm of salivary type arising in the ovaries. PMID- 7483021 TI - [Echocardiography in clinical practice]. PMID- 7483020 TI - Metaplastic breast carcinoma invading chest wall. AB - Metaplastic breast carcinoma refers to a heterogeneous group of neoplasms in which the typical glandular growth pattern of the tumor undergoes metaplasia, either epithelial or stromal. A 59-year-old woman presented with a breast mass that recurred in 1 year and showed invasion of the chest wall. Histological sections of both the tumor and the recurrence showed a tumor composed predominantly of stromal spindle cells with neoplastic epithelial ducts. Squamous metaplasia was seen in some ducts. Immunohistochemical staining showed positive cytokeratin and epithelial membrane antigen staining of the epithelial cells. Smooth muscle actin, S100, and vimentin were diffusely positive in the stromal cells. Electron microscopy of the original lesion showed cells with squamous epithelial and smooth muscle characteristics, and other cells that formed lumens into which microvilli projected. Electron microscopy of the recurrent lesion showed primarily spindle-shaped cells with abundant tonofilaments in the perinuclear cytoplasm, desmosomes with associated tonofilaments, filaments with focal densities, often aligned parallel with the cell membranes, surface attachment plaques, and fragments of basement membrane. Pinocytotic vesicles were rare. These metaplastic cells are derived from myoepithelial cells which are multipotential and able to differentiate into epithelial or stromal cells. PMID- 7483022 TI - [Quality control of echocardiographic examination in a central hospital]. AB - The purpose of this consecutive retrospective study of echocardiographic descriptions and matching case reports was to examine the number of echocardiographic examinations and their consequences for the treatment of the patients. This study was performed in a district hospital in Svendborg, Denmark, during the period from 1.12.93 to 31.03.94 (n = 195). We found that more women than men in the over 69 age group were examined with echocardiographic examination whereas fewer women than men were examined in the 59-69 year-old age group. In 48% of the cases, the actual clinical suspicion was verified by echocardiography; in 40% the echocardiography did not show anything abnormal and in 12% a different diagnosis was found. The echocardiographic examination had a consequence for 29% of the patients regardless the actual clinical suspicion. The frequency of a consequence was the same for the expected and the unexpected finding by echocardiography. Conclusively, this massive rise in the number of performed echocardiographies (n = 51 in 1983; n = 595 in 1993) cannot be reduced by being more restrictive in performing echocardiography for any single clinical suspicion, because all indications for performing echocardiography seem to have equal therapeutic consequences. PMID- 7483023 TI - [Hydatidosis. A survey]. AB - With the increased influx of foreign workers, refugees and immigrants, particularly from endemic areas, we will probably see more patients with hydatid disease, chiefly hydatid disease of the liver caused by E. granulosus. Thus it is important to bring the disease to the attention of the general surgeon. The diagnosis of hydatid disease is primarily made by ultrasound and serological examination. Other tests such as CT, angiography and ERCP are as a rule only important in surgical planning and in complicated cases. Treatment is principally surgical. The choice of surgical procedure should be individualized as much as possible due to the many aspects of the disease. Drainage of residual cavities should be avoided. Preoperative evaluation by ERCP is indicated in cases complicated by biliary communication. Percutaneous drainage can be indicated in cases of recurrence and in cysts of poor accessability. Medical treatment has its place primarily pre- and postoperatively. The preferred drug is albendazole. Mebendazole is now considered obsolete. It is not possible to monitor the effect of treatment with current serological methods. PMID- 7483024 TI - [Laparoscopic cholecystectomy in Denmark. A prospective registration]. AB - Laparoscopic cholecystectomy (LC) was introduced in Denmark in 1991, and a prospective case register was established. All departments performing LC agreed to participate. In 1991-1992, 2,415 patients underwent LC in 44 departments. The median number of procedures was 32 (interquartile range 18-58, range 1-370), performed by a median of four surgeons per department (3-5, 1-23). Two hundred and forty-two patients (10%) had acute cholecystitis. Eighteen point five percent had had an ERCP performed prior to LC. The rate of conversion to open operation was 10.5%, occurring significantly more often in acute cholecystitis (25.6%) than in patients with other indications (8.8%) (p < 0.001). Intraoperative cholangiography was used in 22.4%. The median duration of LC was 90 minutes (70 120, 25-415). The postoperative course was without complications in 90.4%. Laparotomy for complications was necessary in 43 patients (2.0%), mainly because of bile leaks. Twelve patients (0.6%) were treated endoscopically for complications. Bile duct injury occurred in 16 patients (0.66%, 95% CI 0.34 0.99%), including three transsections, one stricture, and 12 minor injuries. Six patients (0.25%, 95% CI 0-0.45%), three of whom had procedure-related complications, died postoperatively. All were > or = 72 years of age. Median time to discharge was two days, while median time to resumed work/normal activity was eight days. A comparison with the number of LC registered in the National Patient Register indicates that reporting is complete. PMID- 7483025 TI - [Outbreak of serogroup C meningococcal disease among teenagers in Randers- preventive measures and examination of the meningococcal carrier conditions]. AB - An outbreak involving 20 cases of serogroup C meningococcal disease, predominantly among teenagers, occurred over a seven-month period in the Randers area of Denmark. The cases were caused by a serogroup C:2a:P1.2 sulphonamide resistant strain. The available evidence was against the transmission being related to particular schools. The outbreak was experienced as three clusters. At two schools involved in the first and the third cluster of the outbreak, 351 students were examined regarding pharyngeal carriage of meningococci, 282 of whom were tested again 17 weeks later; 308 students attending two similar schools in a nearby area were examined once. The majority of strains isolated from group C carriers in the high-risk area were serologically indistinguishable from the outbreak strain (13/14 = 95%), but less often sulphonamide-resistant (5/13 = 38%). In both areas, the overall carrier rate (30%), the overall group C rate (3%) and, the carrier rate for the outbreak strain (1%) were the same. The attack rate for the outbreak strain differed significantly: 1/40 in the high-risk area versus 1/2.500 in the normal risk area. No conditions that might explain this difference were revealed. Immediately after recognition of the first and the third cluster, 780 and 13,300 students, respectively, were vaccinated with meningococcal polysaccharide vaccine A+C. It was concluded that the definition of target groups for vaccination should be liberal, because the "at risk" population may be difficult to recognize at the onset of an outbreak. PMID- 7483026 TI - [Diagnosis and treatment of bacterial vaginosis by general practitioners]. AB - In order to evaluate which diagnostic procedures and treatments general practitioners (GP) used for bacterial vaginosis, and to describe their knowledge of the disease, all the GPs in two counties were asked to fill in a questionnaire. Two hundred and sixty-six (59%) answered. The GP used the criteria of Amsel in 9%, the wet smear criteria in 12%, clue cells in 6%, the amine test in 30%, culture of Gardnerella vaginalis in 29%, 8% only used clinical examination and 6% did not use any criteria. Many examinations were carried out unnecessarily. Sixty-six percent stated that they had a sufficient knowledge of bacterial vaginosis, 36% had knowledge of the criteria of Amsel, 78% of the definition of clue cells and 58% considered culture of Gardnerella vaginalis as being diagnostically useful. Seventy-one percent used metronidazole for treatment. Treatment of the sexual partner was carried out in 33%. In conclusion, we recommend introduction of a more simple criterion than the criteria of Amsel. PMID- 7483027 TI - [Attitude and considerations of sperm donors in relation to insemination]. AB - The object of the study was to examine sperm donors' motivations and emotional reactions to the role as donors and to measure their willingness to provide information about themselves to recipients and offspring. It was designed as a prospective trial in which 26 subjects who were actively functioning as sperm donors at our fertility clinic at the time of investigation were asked to fill in a questionnaire and return it anonymously and voluntarily. Eight percent of the donors stated purely altruistic motivation, 32% purely financial and 60% a combination of both. The acceptance rate for providing non-identifying, phenotypic, descriptive information to recipients is 76%, for providing medical information it is 60% and the provision of psychosocial information is accepted by 28% to 40% depending on the item. Twenty percent of donors are willing to continue donation if the present rules of anonymity are revoked, 20% were undecided. In conclusion, most donors do not seem to feel any close relationship to donor offspring and at least 60% found anonymity to be essential for their continued functioning as donors. PMID- 7483028 TI - [Surgical treatment of "mushroom poisoning"]. PMID- 7483029 TI - [Heart massage. Liver rupture and thrombolysis]. AB - Complications to external cardiac massage with special regard to liver damage are shortly reviewed. A case story, in which a young healthy woman suffered rupture of the liver followed by hypovolaemic shock after a few minutes of cardiac massage and subsequent thrombolysis, is presented. It is discussed whether the common use of thrombolytic treatment will cause more cases of clinically important haemorrhage after external cardiac massage. PMID- 7483030 TI - [Telomerase. The enzyme that renders the cell immortal]. PMID- 7483031 TI - [Paracetamol poisoning]. PMID- 7483032 TI - [Improved prognosis in breast cancer]. PMID- 7483033 TI - [A combiphasic oral contraceptive containing ethinylestradiol and desogestrel]. PMID- 7483034 TI - [Prehospital treatment of heart arrest]. PMID- 7483035 TI - [Guidelines for resuscitation of adult patient. Danish Resuscitation Council]. PMID- 7483036 TI - [Heart arrest outside hospital. Survival, importance of early defibrillation]. PMID- 7483038 TI - [Single-session bilateral hip alloplasty. A simultaneous procedure in 79 patients]. AB - The aim of the study was to evaluate the per- and postoperative complications in 79 consecutively operated selected patients with bilateral hip disease who had bilateral hip replacements performed in one session. It was carried out as a retrospective examination of the case records, a questionnaire and clinical and radiological follow-up of 41 patients after a median of 7.5 years. No serious complications were registered peroperatively, and the duration of anaesthesia was on average three and a half hours. Eight patients were treated for early postoperative complications. The average hospitalization time was 15 days. Forty one patients were followed up at a minimum of five years postoperatively. All patients were fully satisfied with the result, and there had been no cases of dislocation or reoperation. The average hip function index had been 7.2 preoperatively, and was at follow-up found to be 16.7. Radiological examination showed signs of looseness in 12 of the 82 examined hips. It is concluded that in selected patients with bilateral hip disease necessitating bilateral hip replacement the two operations may with benefit be carried out in one session. PMID- 7483039 TI - [Occurrence of toxoplasmosis in pregnant women in Denmark. A study of 5.402 pregnant women]. AB - Consecutive serum samples from 5402 women were analyzed for specific IgG and IgM Toxoplasma gondii antibodies by enzyme immunosorbent assay. The prevalence of IgG antibodies was 27.4%. The seroconversion rate was estimated to be 1.16% per year (SE = 0.0017) in all age groups, consistent with an incidence of acute toxoplasmosis in pregnant women of 0.65%. Based on theses figures, the estimated number of congenitally infected cases are discussed. There was no difference in the prevalence of antibodies in women from rural or urban areas, or from different parts of Denmark. PMID- 7483037 TI - [Kaposi sarcoma. An epidemiological perspective]. AB - Prior to the 1980s, Kaposi's sarcoma was a rare tumour diagnosed three to four times more frequently among men than women. It was primarily seen among elderly men of Mediterranean or Jewish descent, in well-defined areas of Central Africa, or more scattered as individual cases with underlying immunosuppression. Geographical restrictions and suggested associations with certain HLA-types gave rise to early speculations of a genetic component involved in its etiology. With the AIDS epidemic, the epidemiology of Kaposi's sarcoma changed drastically. Although diagnosed among AIDS patients that are transfusion recipients, intravenous drug users and haemophiliacs, Kaposi's sarcoma is primarily found in homosexual men with AIDS among whom the risk has increased to 100,000 compared to the general population. Specific behaviours linked to homosexual men have been sought to explain this relationship, but accumulating evidence favours the involvement of an infectious agent in the etiology of Kaposi's sarcoma. The most promising candidate today is a possible new herpesvirus, similar to but distinct from Herpes-virus saimiri and Epstein-Barr virus. PMID- 7483041 TI - [Organization of clinical research at a hospital department]. PMID- 7483040 TI - [Psychiatric morbidity in connection with surgical treatment of epilepsy. A short term follow-up of patients with amygdalohippocampectomy]. AB - The aim of the study was to examine the occurrence of psychiatric disorders in epilepsy patients who had received surgical treatment, especially amygdalohippocampectomy (AHE), for the relief of medically intractable seizures. Forty-seven subjects, treated during the period 1987-1991 in the Danish epilepsy surgery programme (EPIKIR), entered a retrospective interview study. Of these, 37 had undergone AHE. Preoperative psychiatric morbidity was assessed through interview and available case notes, including a routine psychiatric interview. Postoperative psychiatric morbidity was assessed by the use of the Present State Examination. A total of six subjects (five AHE subjects) developed depressive disorders of various duration and severity after operation. In three subjects this occurred "de novo". No paranoid-hallucinatory psychoses developed within the follow-up period (a minimum of one year), and the presence of psychiatric disorders could not be associated with either lateralization of cerebral dominance of histopathological findings. Thus, depression appears to be the most frequent psychiatric problem following epilepsy surgery. Although the present study mainly deals with AHE, this finding is in accordance with the results of recent findings concerning anterior temporal lobe resection. PMID- 7483042 TI - [Molybdenum cofactor deficiency. Clinical symptoms and diagnostic strategy]. AB - The clinical, neuroradiological and biochemical findings in two siblings with molybdenum cofactor deficiency are presented. A search for this deficiency is advocated in each case of unexplained refractory neonatal convulsions. Early diagnosis is prompted by the often rapid fatal outcome and the availability of methods for prenatal diagnosis. Diagnosis may be missed or delayed on standard metabolic screening for several reasons discussed. Magnetic resonance imaging in this condition seems to be rather characteristic. PMID- 7483043 TI - [Improvement of milk substitutes by addition of special fatty acids]. PMID- 7483044 TI - [Carbohydrate deficient transferrin--reliability of the test?]. PMID- 7483045 TI - [Molecular biology that disappeared]. PMID- 7483046 TI - [Drug interactions in patients admitted to a department of internal medicine]. PMID- 7483047 TI - [Scintigraphy of abdominal bleeding--a diagnostic alternative?]. PMID- 7483048 TI - [Scintigraphy of bleeding. The diagnostic value based on a 10-year period]. AB - The purpose was to examine the diagnostic value of 99mTc-labelled red blood cell scintigraphy for detection of gastrointestinal haemorrhage. In a retrospective investigation 85 patient files over a 10 year period (1.1.1984-31.12.1993) were evaluated. Data regarding the patients' clinical condition, number of blood transfusions, findings with angiography, gastroscopy, colonoscopy, proctoscopy, operation, x-ray of colon and autopsy were registered. A bleeding site was localized in 52% of the scintigraphies. Among these patients the scintigraphic localization was confirmed or made "very possible" in 65% by operation, autopsy or one of the other mentioned diagnostic modalities. Among the 25 operated patients the localization of the bleeding site was confirmed in 19 patients. In 40 patients with a "negative" scintigraphy a bleeding site was found in 10 patients 1-18 days after the scintigraphy by operation or one of the listed diagnostic modalities. It is concluded that 99mTc-labelled red blood cell scintigraphy is a valuable diagnostic tool for identification of acute gastrointestinal haemorrhage. Operation after scintigraphic guidance is well indicated. A "negative" scintigraphy can only exclude ongoing bleeding from the gastrointestinal tract. PMID- 7483049 TI - [Renal function during treatment with angiotensin converting enzyme inhibitors]. AB - The use of ACE-inhibitors has increased greatly during the last years. They were first used in treating hypertension, but nowadays cardiac diseases, mainly cardiac failure, are common indications. This means that the drugs are used in the treatment of more elderly patients who often have generalised atherosclerosis. This means that the patients must be controlled more often after initiation of treatment, especially concerning kidney function, since treatment with ACE-inhibitors can cause pronounced changes in renal haemodynamics and kidney function. This review focuses on the effects of ACE-inhibitors on renal haemodynamics and kidney function, which may be positive, with preservation of kidney function in diabetic and other chronic nephropathy, or negative, for example in cases with atherosclerotic stenosis of large or small renal arteries. It is concluded, that in cases of diabetic nephropathy an ACE-inhibitor is the "drug of choice" for treatment of hypertension. Furthermore the ACE-inhibitors seem to reduce the rate of deterioration of renal function and proteinuria in other kidney diseases. It is emphasized, that during treatment with ACE inhibitors kidney function must be controlled before and following one to two weeks of treatment, if the dose is changed and in all cases following two to three months of treatment. Special attention should be given to patients with atherosclerotic manifestations e.g. angina. PMID- 7483050 TI - [Renal function during treatment of chronic renal failure with angiotensin converting enzyme inhibitors]. AB - The effect on kidney function fo treatment of cardiac failure with ACE-inhibitors was examined retrospectively in a material of 87 consecutive patients. Furthermore, it was evaluated whether concomitant treatment with diuretics or existing generalised atherosclerosis as indicated by ongoing treatment with nitrates could be a risk factor concerning reduction of kidney function. In 11.9% of the patients an increase in S-creatinine of > 30% was observed during the first weeks of treatment. It was only necessary to stop treatment in two of these patients. In the remainder S-creatinine decreased again during ongoing treatment. In another 10.7% of patients an increase of 20-30% in S-creatinine was observed. Seventy-two point six percent of the patients had unchanged kidney function during treatment with an ACE-inhibitor. Ongoing treatment with diuretics did not seem to be a risk factor for developing reduced kidney function, whereas significantly more patients on treatment with nitrates, indicating generalised atherosclerosis, developed reduced kidney function during treatment with ACE inhibitors. It is recommended to control kidney function before, one to two weeks and two to three months following initiation of treatment with ACE-inhibitors and to pay special attention to patients with generalised atherosclerosis. PMID- 7483051 TI - [Traumatic myositis ossificans. Posttraumatic non-neoplastic heterotopic ossification]. AB - Myositis ossificans traumatica (MOT) is a nonneoplastic, heterotopic ossification of soft tissues i.e. skeletal muscle, tendons, aponeuroses and fascia. It is often encountered in young male athletes participating in contact sports as a result of a single or repeated contusion. MOT tends to be solitary, localized and well circumscribed with a self-limited growth potential that may culminate in regression. The pathogenesis of MOT is still enigmatic. Recent animal experiments have led to a theory that mesenchymal connective tissue cells, undergo metaplasia induced by trauma and probably osteogenic proteins, to fibroblasts and osteoblasts. These cells deposit and structure osteoid centripetally in the lesion. As the lesion matures, cancellous bone develops into mature, lamellar bone in the periphery of the lesion. In its earlier stages MOT is easily cytologically and radiologically confused with osteogenic sarcoma. The management of MOT is largely conservative and the principles are of considerable value to physicians and physiotherapists engaged in the treatment of sports injuries. This article reviews the various forms of myositis ossificans as well as the pathology, diagnosis and treatment options. PMID- 7483052 TI - [Prediction of bleeding risks and death in cirrhosis based on hemodynamic and metabolic variables. A study of patients with esophageal varices without previous bleeding]. AB - As identification of patients with liver cirrhosis who are at risk of bleeding or death is essential in terms of prophylaxis, we have determined the prognostic influence of various patient characteristics on risk of bleeding and death. Fifty five cirrhotic patients with oesophageal varices without previous bleeding were included in the study and followed up after a medium observation period of 446 days (range:5-1211 days). A total of 55 clinical, biochemical, haemodynamic, and endoscopic variables were classified as either systemic haemodynamic, portal haemodynamic, or metabolic. Using univariate analysis the following variables showed a significant relation to an increased risk of bleeding or death: high plasma volume (p < 0.02), high azygos blood flow (p < 0.004), high hepatic venous pressure gradient (p < 0.02), marked prominence of varices (p < 0.05), poor nutritional status (p < 0.0001), low clotting factor 2, 7, 10 (p < 0.002), low incapacitation index (p < 0.004), low serum albumin (p < 0.005), high bilirubin (p = 0.05), high alkaline phosphatases (p < 0.02), low arterial oxygen saturation (p = 0.02), and encephalopathy (p < 0.007). In a Cox regression model poor nutritional status (p < 0.00005), serum bilirubin (p < 0.001), short central circulation time (p < 0.03), low serum albumin (p < 0.02), and low index of clotting factor 2, 7, 10 (p < 0.05) were independently associated with higher risk. In conclusion, the results support the prognostic value of metabolic variables as earlier described. The prognostic significance of central circulation time stresses the importance of the hyperdynamic systemic circulation in the increased risk.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7483053 TI - [Interobserver variation in the evaluation of neurobiological findings: observer dependent and patient-dependent variables]. PMID- 7483054 TI - [Subcutaneous fat necrosis with hypercalcemia]. AB - A case of subcutaneous fat necrosis (SFN) is presented. The infant developed late onset hypercalcaemia of severe degree. Calcitonin failed to normalize the hypercalcaemia. Glucocorticoids and withdrawal of dietary calcium and vitamin D are the treatment of choice. Like hypercalcaemia in association with other granulomatous diseases, SFN is possibly secondary to locally produced 1,25 dihydroxy-cholecalciferol. Our results were not able to confirm this theory. Infants with SFN should have calcium levels monitored closely for months. PMID- 7483055 TI - [Acute pancreatitis--induced by 5-aminosalicylic acid or an extraintestinal manifestation of ulcerative colitis?]. AB - 5-aminosalicylic acid has been reported to cause acute pancreatitis. We report the first Scandinavian case. A 27 year-old woman with a three month history of ulcerative colitis treated with Mesalazine developed acute abdominal pain. Serum and urine amylase were raised, and ultrasonographic and computed tomography scan showed oedema of the pancreas. Immediately following withdrawal of the drug, serum and urine amylase returned to normal. We discuss the possibility of acute pancreatitis as an extraintestinal manifestation of ulcerative colitis. PMID- 7483056 TI - [Local recurrence after surgery for rectal cancer. Could surgeons do it better?]. PMID- 7483057 TI - [A restrictive antibiotic policy in practice]. PMID- 7483058 TI - [Hepatorenal syndrome]. PMID- 7483059 TI - [Postoperative consequences after male sterilization]. PMID- 7483060 TI - [Chronic lung disease in premature infants]. PMID- 7483062 TI - [Free oxygen radicals during transplantation surgery]. AB - Free oxygen radicals are a highly reactive species of molecules characterized by an unpaired electron in the outer electron ring. They are formed continuously in the organism as part of enzymatic reactions, or as byproducts of oxidation processes. Antioxidants are able to eliminate free oxygen radicals, but during ischaemic reperfusion, the capacity of these systems is exceeded resulting in tissue injury, cell necrosis, destruction of enzymes and DNA-damage. Free oxygen radicals thereby play a major role in the tissue injury seen in organs after transplantation surgery and aorta operations. When the endogenous antioxidative capacity is exceeded, protection of lipid membranes against attack by free oxygen radicals by antioxidant supplementation is an alternative strategy for reduction of tissue injury. Since antioxidants are atoxic, they could also be of importance even in small operations, where the tissue blood supply is often periodically cut off. PMID- 7483063 TI - [A local course on conventional and laparoscopic surgical techniques. A supplement to surgical education]. AB - In order to increase the possibilities for learning accurate surgical technique in gastrointestinal surgery, we have established a local course for operative training of the registrars. We used organs from pigs--stomach, intestines and liver/gallbladder. They were collected from the local abattoir with assistance from a vet. Conventional gut anastomosis and gastroenteroanastomosis were performed with "two-layer" and "one-layer" techniques. Laparoscopic cholecystectomy was performed with the liver-gallbladder organ block placed in a "black box", in which the organ was perfused with red fluid--a Pulsatile Organ Perfusion System. The registrars' evaluations of the course were very positive. They all thought that the course gave them opportunities for surgical training that were not available in the busy daily routine. The course now forms part of the education programme for surgical registrars in our department. PMID- 7483061 TI - [Multiple cystic pulmonary changes in a patient with previous bronchopulmonary dysplasia]. AB - This report describes multiple pulmonary air cysts in a twelve year-old girl. The pathogenesis is discussed, taking into consideration the possible role of prematurity and bronchopulmonary dysplasia. PMID- 7483064 TI - [Overweight and urinary incontinence in women]. AB - The aim was to study the possible role of obesity in adult female urinary incontinence (UI) etiology. A random population sample of 3114 women aged 30-59 were sent a postal questionnaire concerning urinary incontinence and, among other things, body weight and height. The overall response rate was 85%, and the present analysis comprises 2589 women who gave information about their body weight and height. The period prevalence of all, stress, urge, and mixed stress and urge urinary incontinence (UI) was 17%, 15%, 9% and 7%, respectively. The mean body mass index (BMI) was 22.7 kg/m2. Irrespective of other risk indicators, BMI was positively associated with UI prevalence (OR 1.07 per BMI unit, p < 0.0001). BMI interacted with childbirth in predicting stress UI prevalence, with cystitis in predicting urge UI, and with both in predicting mixed UI. Stress UI turned out to be the UI type most closely associated with BMI. PMID- 7483065 TI - [Acoustic neurinomas/vestibular schwannomas. Diagnostic evidence vs. surgery]. AB - +-f having vestibular schwannoma (VS) was conducted in a series of 123 patients (127 tumours), over a 20 year-period from 1973 to 1993. Mean follow up period was 3.4 years, mean annual growth rate was 3.2 mm/year. Tumour growth was observed in 90 patients (94 tumours, 74%), no growth was seen in 23 patients (23 tumours, 18%) and negative tumour growth in 10 patients (10 tumours, 8%). Surgery due to tumour growth was performed in 35 patients (35 tumours, 28%). Seven patients (seven tumours, 6%) were treated with gamma radiation and/or shunt insertion. Seven patients (6%) died of brainstem herniation induced by tumour compression. Nine patients (7%) died of non tumour related causes. Twenty-eight patients were classified as candidates for hearing preservation surgery and 21 patients (75%) lost their candidature during the observation period due to tumour growth and/or deterioration of hearing. The results may limit indications for allocation of patients with VS to the "wait and see" group. PMID- 7483066 TI - [Acute colonic ileus caused by left-sided colorectal cancer. A randomized trial of emergency ostomy versus resection]. AB - Staged resection (group T) versus acute resection (group R) for curative purpose was compared in a randomized study of 121 patients presenting with signs of left sided obstructive colorectal tumours during emergency surgery from 1978 to 1993. Patients with distant spread were excluded. Transversostomy was done in 58 and resection without immediate anastomosis in 56. Duration of emergency surgery was shorter, blood transfusions fever and wound infections less frequent in T compared to R, but postoperative mortality was similar (eight patients in each group). The diagnosis of tumour was wrong in 11 patients in T and six in R. The proportion of patients surviving the second stage curative resection in T without a permanent colostomy (32/35) was higher than after acute resection (36/50) in spite of six patients having anastomotic surgery (Coloshield) at the time of acute resection in R. Days spent in hospital were less in R. Overall recurrence rates and survival rates were similar in T and R. In conclusion, no major advantage besides shorter hospital stay could be demonstrated by acute resection without simultaneous anastomosis compared to the traditional 3-stage procedure, which on the other hand carried a much smaller risk of a permanent colostomy. The latter should therefore serve as a control in a prospective evaluation of emergency resection with simultaneous anastomosis. PMID- 7483067 TI - [Pediatric evaluation of children with heart murmurs]. AB - In order to analyse the non-specialized paediatrician's ability to evaluate children with a heart murmur and the consequences of this evaluation for the need of further investigations, we made a prospective and consecutive study concerning 100 children who were referred for initial evaluation of a heart murmur. Based on the clinical evaluation, the children were assigned to one of the three categories of "no heart disease" "heart disease" or "definite heart disease". Within two months each child underwent a complete colour Doppler echocardiographic study. Except for one child with an insignificant peripheral pulmonary artery stenosis, 53% of the children were correctly diagnosed as having no heart disease. We conclude that clinical evaluation without laboratory tests performed by paediatricians without any special training in cardiology could correctly acquit half of the children of heart disease, with no need for further investigation. PMID- 7483068 TI - [Indications for the use of blood. A quality assurance project]. PMID- 7483069 TI - [Responsibility of the Ugeskrift for quality research]. PMID- 7483070 TI - [Is home childbirth safe? A meta-analysis is considered]. PMID- 7483072 TI - [Caution rules when prescribing valproate]. PMID- 7483071 TI - [Oral gamma-linoleic acid (Epogam) in the treatment of atopic dermatitis]. PMID- 7483073 TI - [Thyroid gland surgery--who should perform it?]. PMID- 7483074 TI - [Thyroid gland surgery in a teaching department of parenchymal surgery. Quality control and perspectives based on a 10-year material]. AB - The complications after surgery for benign thyroid disease in a non-specialized department were evaluated in a retrospective study. Furthermore it was evaluated whether the new recommendations of The National Board of Health for referral of patients in need of thyroid surgery would influence the rate of operations and complications in the ward. Three hundred and seventy-four patients were operated on. The complication rate was at the same level as in earlier Danish reports. Permanent unilateral palsy of the recurrent laryngeal nerve was registered in 2.1% of the patients (i.e. in 1.4% of "nerves at risk") in patients with primary benign goitre. The complication rate was slightly, but not significantly, higher after operations for recurrent goitre. Hypoparathyroidism occurred in 1.8%, infection in 3% and haemorrhage in 5.3% of the patients. If the recommendations of The National Board of Health for referral of patients were applied the complication rate would decrease, but not to a level of statistical significance. The number of operations would decrease by 20%. From the findings it is argued that not more than one or two hospitals in a county should perform thyroid operations and that not every surgeon should be trained in thyroid surgery. PMID- 7483075 TI - [Complications after surgical treatment of malign thyroid diseases]. AB - Eighty-eight patients with thyroid malignancy seen at the Rigshospital, Copenhagen during the period 1.5.1989-1.5.1994 were retrospectively studied. There were 64 females and 24 males. The average age was 54 years. Seventy-nine patients were operated (six with subtotal and 73 with total thyroidectomy). Thirty-six (46%) had had previous thyroid operations and 12 of these patients had preoperative fine-needle aspiration biopsies performed. The second operation thyroidectomy specimen contained carcinoma in 14 (39%) of the 36 patients. Forty nine patients (62%) had different types of modified neck dissection performed under the operation and 31 (39%) of the operated patients had lymph-node metastases. Among the patients who were operated at the department there were no accidental permanent recurrent nerve paralysis. Among the 36 patients who were operated at other departments there were four (11%) accidental recurrent nerve paralysis from the primary operation. Twenty (25%) of the 79 operated patients developed permanent hypoparathyroidism. PMID- 7483076 TI - [Surgical treatment of goiter at a central hospital. A consecutive study with special emphasis on surgical complications]. AB - The aim of this study was to analyse the frequency of permanent and transient complications following thyroid gland surgery. During a period of four years, from 1991 to 1995, 129 patients were treated operatively for thyroid disease. Sixteen patients were operated for thyrotoxicosis and 113 for nontoxic goitre. Nine patients underwent reoperation for recurrent goitre. Thyroid malignancy was found in two patients. The frequency of permanent complications was 0.7%. Transient unilateral pareses developed postoperatively in two patients. There were no permanent vocal cord paralyses. One patient developed permanent hypoparathyroidism. Five patients had transient postoperative hypocalcaemia requiring calcium supplements. Other complications were haemorrhage (3.8%), wound infection (0.7%) and other postoperative complications (1.5%). It is shown that uncomplicated thyroid surgery may be performed at a district hospital that has a special interest in patient evaluation, indications for surgery and up to date surgical technique. PMID- 7483077 TI - [Alcoholic ketoacidosis]. AB - Alcoholic ketoacidosis is an acute metabolic disorder that occurs in ethanol abusers who because of gastritis or other concurrent illness stop eating and drinking and vomit repeatedly. This causes dehydration and ketoacidosis with little or no hyperglycaemia and glucosuria. A case of severe alcoholic ketoacidosis is presented and the literature on alcoholic ketoacidosis is reviewed. PMID- 7483078 TI - [Amputation and arterial reconstruction in peripheral occlusive arterial diseases in Denmark 1977-1993]. AB - Information was retrieved from the national patient register (Landspatientregisteret), that contains information on all admissions to Danish hospitals 1977-1993. The number of admissions during which a diagnosis of occlusive arterial disease (OAD) of the lower extremities was made and an amputation performed increased from 1290 in 1977 to a maximum of 1609 in 1983 (p < 0.001). From 1987 on the number decreased almost 30% to 1111 in 1993 (p < 0.001). During the period the number of admissions during which arterial reconstruction for OAD were performed increased steadily from 963 in 1977 to 2637 in 1993 (p < 0.001). The result may indicate that vascular surgery is effective on a national scale in preventing amputations for OAD. PMID- 7483079 TI - [Menstrual cycle and surgery of breast cancer. Point of time for the surgery of primary breast cancer in connection with menstrual cycle is without prognostic significance]. AB - From 1977 to 1989 6488 patients under fifty years with primary breast cancer were registered in the nationwide Danish Breast Cancer Cooperative Group (DBCG). Among these information on last menstrual period prior to surgery was available in 1635 cases which constitute the study group of the present analysis. The group was representative of the total group with regard to prognostic factors and survival. In the study group time of surgery in relation to last menstrual period was found to have no influence on five- and ten year survival. PMID- 7483081 TI - [Morning-after pill--abortion]. PMID- 7483082 TI - [Registration of bladder tumors--the bath-water and the child]. PMID- 7483083 TI - [Cataract surgery]. PMID- 7483084 TI - [Results after cataract surgery]. AB - This study reports the short term surgical results in a consecutive series of 1473 patients undergoing cataract surgery (60% planned extracapsular cataract extraction (ECCE) and 40% phacoemulsification (KPE) on an outpatient basis in a major university eye clinic. Preoperatively, 80% of all cases had a corrected visual acuity of 0.3 (6/18) or lower. On the first day after surgery, 51% of all patients had a corrected visual acuity of 0.5 (6/12) or better and 9% had a visual acuity of 1.0 (20/20) or better. The unaided visual acuity was 0.5 (6/12) or better in 20% of all patients (78% of these after KPE). The average postoperative refraction was -0.43 D (+/- 1.03 D SD, range -6.00 D to +5.00 D). A significant improvement in refractive error was achieved in many patients. The mean intraocular lens (IOL) power prediction error was found to be +0.04 D (+/- 0.84 D). The surgically induced astigmatism (vector analysis) was found to be +4.91 D (+/- 2.08) in the ECCE cases and +1.37 D (+/- 1.38) in the KPE cases (p < 0.001). Capsular or zonular rupture and/or vitreous loss occurred in 43 (2.9%) of which 26 (1.8%) received an anterior chamber lens rather than the intended posterior chamber lens. The results of this study document the favourable results of modern cataract surgery. PMID- 7483080 TI - [Parvovirus B19 infection as the cause of hepatitis and neutrophil granulocytosis in a 20-year old woman]. AB - A case of Parvovirus B19 infection (erythema infectiosum) in a 20 year old woman is presented. The patient presented with fever, arthritis in one knee, neutrophil granulocytosis and biochemical evidence of hepatitis. Serological evidence of Parvovirus B19 infection was found as the only explanation of the clinical picture. Hepatitis was due to Parvovirus B19 infection as there was no serological evidence of EBV or CMV reactivation. Neutrophil granulocytosis and thrombocytosis were found and were probably due to an active bone marrow in the recovery phase of bone marrow aplasia. PMID- 7483086 TI - [Meningeal carcinomatosis in solid tumors]. AB - Meningeal carcinomatosis (MC) originating from non-haematological tumours is a late event in the course of a malignant disease. MC must be suspected in patients with disperse neurological symptoms from at least two of the following three parts of the central nervous system: the brain, the cranial nerves, and the spinal cord, and if a discrepancy between symptoms and objective findings is found. A suspicion of MC must be confirmed by lumbar puncture and/or MR with Gadolinium in patients where a confirmation of MC will lead to active therapy. Therapy depends on tumour type, performance status of the patient, and present status of the malignant disease. Patients with good performance status and with a chemo-sensitive tumour are offered a combination of radiotherapy and chemotherapy. Median survival is one to three months and active therapy does not prolong survival with certainty. Therefore, therapy primarily aims at an improvement of quality of life for the patient. PMID- 7483085 TI - [Quantification of quality]. PMID- 7483088 TI - [HIV testing in Denmark in 1993: were the tests relevant?]. AB - In order to describe HIV-testing activity among adults in Denmark, a cross sectional study was designed using an anonymous questionnaire, which was forwarded with all HIV-test results in a given two-week period in 1993. Notified first-time identified HIV-positive cases were used for comparisons. The response rate was 50%. A total of 996 women and 834 men were included, among whom 68% of men and 62% of women were classified as having a behaviour that put them at increased risk for acquiring HIV, p < 0.02. Women were younger than men, p < 0.001. Around 1/3 of both men and women had previously been HIV-tested, a proportion that increased with increasing risk for HIV, p < 0.00001 for trend. HIV-tested adults were younger and less frequently living in the Copenhagen area than the first-time identified HIV-infected persons reported to the mandatory national HIV-surveillance system. The overall HIV prevalence was 0.3%, being 0.5% among men and 0.2% among women. Estimates showed that the yearly number of first time identified HIV-infections among tested homo/bisexual men per 10(5) men were 100 times as high as among the heterosexual population. Also, 12% of Danish homo/bisexual men had an HIV test per year, a proportion that was 2.3% among adult women and 1.8% among adult heterosexual men without i.v. drug use. The yearly test-activity decreases with increasing age among both men and women. In conclusion, studies among the HIV-tested population along with results from systematic surveillance of first-time identified HIV-infections are useful in order to estimate the frequency and the relevancy of HIV testing in the entire population. Also, these results may help when planning national information campaigns. PMID- 7483087 TI - [The effect of ranitidine on postoperative monocyte and neutrophil granulocyte function]. AB - The histamine H-2-receptor antagonist ranitidine hydrochloride has been shown to alleviate trauma-, blood transfusion- and sepsis-induced immunosuppression. We evaluated the effect of ranitidine on the postoperative impairment of monocyte and neutrophil function in 24 patients undergoing major elective abdominal surgery. The patients were randomized to receive postoperative adjuvant treatment with ranitidine hydrochloride (100 mg) administered intravenously twice daily for four days, followed by oral ranitidine hydrochloride (150 mg) administered twice daily for five days (n = 11), or no adjuvant treatment (n = 13). Blood monocyte and neutrophil chemotaxis and chemiluminescence were analyzed before the operation and on post-operative days one, three and nine. Monocyte chemotaxis to C5a in the 13 control patients was significantly decreased on day one compared to day 0. Chemotaxis in the 11 ranitidine-treated patients increased significantly from day 0 to day one (p < .01 between groups). Neutrophil chemiluminescence to zymosan and N-f-methionyl-leucylphenylalanine was significantly increased in control patients on day one compared to day 0 (p < .05), while ranitidine reduced chemiluminescence to zymosan insignificantly on day one (p < .07 between groups). Five of the 13 control patients developed postoperative infectious complications, which were related to decreased monocyte chemotaxis to C5a and increased neutrophil chemiluminescence to zymosan when compared to noninfected patients. A significant difference (P < .05) in chemiluminescence to zymosan between infected and noninfected control patients was observed on day three, before clinical signs of infectious disease could be detected. There were no infectious complications in ranitidine-treated patients. These results support previous studies on the effect of ranitidine in reducing postoperative immunosuppression. PMID- 7483089 TI - [Vermillionectomy]. AB - Several external factors such as sunlight and tobacco smoking may cause changes in the epithelium of the vermillion border, especially of the lower lip, resulting in a dry and crusty epithelium often accompanied by painful ulcerations. Some of these patients will eventually develop lip cancer in the vermillion border. We would like to present a consecutive series of 25 patients treated with vermillionectomy. It is a very good and efficient treatment which may be the solution to a long standing problem. The functional and cosmetic results are very good. PMID- 7483090 TI - [Statistical study of workload at a birthing unit]. AB - The work consists of a statistical study of the workload on a labour ward. This included a study of the admission/birth frequency and an investigation of the factors that influenced the process of admissions and the time spent on the ward. Also the relation between the number of midwives and women in labour present on the ward was investigated. It was found that the variation in the number of spontaneous admissions/births could be described reasonably well by a Poisson distribution. A simple statistical model was proposed to calculate the number of midwives necessary at any given birth frequency. For a specific choice of parameters the model fitted the actual distribution very well. The relative frequency of complications was highest for women with no previous births and smallest for women with one previous birth. The time spent on the labour ward depended significantly on the number of times the woman had given birth before and on whether the present delivery was complicated. PMID- 7483091 TI - [Spontaneous regression of pulmonary metastases after nephrectomy for renal cell carcinoma]. AB - A case of spontaneous regression of multiple pulmonary metastases from renal cell carcinoma in a 58-year-old woman after a right radical nephrectomy is reported. Two years after the operation the patient is in good general condition and has gained weight. The clinical aspects of this phenomenon are discussed. PMID- 7483093 TI - [Home delivery and considering the risks]. PMID- 7483092 TI - [Avulsion fracture of the ischial tuberosity. A rare lesion whose early diagnosis and correct treatment may prevent late sequelae]. AB - Avulsion fractures of the ischial tuberosity are often mistaken for sprains or a muscle tears. We present two cases of displaced fractures with a diagnostic delay of eighteen months and two years, respectively. Both cases were treated conservatively and resulted in discomfort on sitting and in one case persistent functional disability. Since early diagnosis is important for a successful treatment, an anteroposterior radiograph of the pelvis is recommended in patients presenting a history and symptoms indicating a lesion in the posterior aspect of the upper thigh. Accumulated experience indicates, that acute displaced fractures should be treated by open reduction and internal fixation. PMID- 7483094 TI - [MR-scanners in Denmark]. PMID- 7483095 TI - [In vino veritas?]. PMID- 7483097 TI - [Hepatic vein catheterization. Use in clinical practice and research]. PMID- 7483096 TI - [Treatment of esophageal varices in cirrhosis]. PMID- 7483098 TI - [Portal hypertensive gastropathy]. AB - Portal hypertensive gastropathy (PHG) as defined by congestive changes in the gastric mucosa owing to increased portal pressure, was first described about ten years ago. Whereas definition and grading of severity are still under debate, there is general agreement that PHG is a new clinical entity. PHG is present in 50-80% of patients with liver cirrhosis. PHG is a major cause of upper gastrointestinal bleeding in patients with portal hypertension (25-90% depending on severity). Presence of portal hypertension is a prerequisite for the development of PHG, and reduction of portal pressure and splanchnic blood flow with beta-adrenergic blockers has shown promising results, but an established medical treatment of PHG does not exist. Trials with new vasoactive drugs are awaited. PMID- 7483099 TI - [Total hip alloplasty using the Muller method. A six and a half year follow-up study]. AB - From January 1984 to December 1986, 383 total hip arthroplasties were performed in 353 patients. Median age at operation was 69 (24-85) years. At follow-up median 78 (62-102) months after operation, 261 patients with 278 arthroplasties were still alive. A questionnaire was sent to all living patients, 258 patients (99 per cent) replied. Fifty-seven patients with 61 arthroplasties complained of pain and were called in for a check-up including x-ray. Including the dead patients a total of six arthroplasties had been revised, two because of aseptic loosening of the cup and three because of loosening of both components. One had been revised because of deep infection. In the patients called in for check-up seven were suspected of loosening of the stem and one of loosening of the cup. Survival analysis according to Kaplan-Meier showed a survival rate of the prosthesis of 98.5 per cent at 61/2 years. It is concluded that the results after total hip arthroplasties with the straight stem Muller prosthesis are satisfactory. The patients from this study will be followed prospectively from now on to investigate the long-term survival of this type of prosthesis. PMID- 7483100 TI - [Back problems in conscripts. A 10-year follow-up]. AB - One hundred and twenty-three conscripts called up for national service in 1979-80 were re-examined 10 years after call up with a questionnaire identical to one answered at the time of call up. The study group consisted of three subgroups. Group A had been rejected for service because of back problems (back-rejects). Group B were conscripts with back symptoms at the time of call up, but all completed their national service (non back-rejects). Group C did not have back symptoms at the time of call up. The back-rejects had a significantly higher incidence of: back pain; radiating pain to lower extremities; contacts (hospital, G. P., radiographs etc.) and sick leave from job as compared to group B and C. Out of 19 conscripts with radiographically verified Mb. Scheuermann, 89% had back symptoms at the time of follow-up including all with lumbar and thoracolumbar Mb. Scheuermann. All conscripts in this study had a job at follow-up. It is recommended that the criteria for rejection of conscripts in Denmark concerning back symptoms and Mb. Scheuermann remains unchanged. PMID- 7483101 TI - [Mandatory anonymous HIV-surveillance in Denmark. Experience with the new notification system]. AB - A newly implemented mandatory, anonymous HIV surveillance system in Denmark is described. The system is based on a serially numbered form that is circulated among the laboratories, the physicians, and the national surveillance unit. All laboratories doing confirmatory tests for HIV antibodies must send a report form along with any positive test result. Before forwarding the form to the physician, the laboratory is to fill in the top part of the form with its own name, the name and address of the requesting physician and the date of testing. The back copy must be send to the surveillance unit. The Danish physicians are by law mandated to fill in the form with data on the patient and then send the original to the surveillance unit. The two copies are for the patient's file and for the physicians' own centralized filing system. After a revision of the system in 1992, a response rate of 95% was obtained during the first year. An increasing number and proportion of first-time identified HIV-infections in Denmark are acquired heterosexually, many of these infections are identified among foreigners. The newly developed mandatory HIV surveillance system is based on anonymous reporting but it allows routine evaluation of the completeness of reporting as well as nonresponding physicians to be reminded of their duty to report. Also, the system allows nested studies to be carried out among selected HIV-infected subpopulations of special research interest without compromising the anonymity of the patient. PMID- 7483102 TI - [Fatal cases of poisonings among drug addicts in the county of Funen in 1993]. AB - This study includes death from poisoning among drug addicts in 1993 in the county of Funen, in all 24. The development during the last five years shows an increase, especially in the largest city in the county. The drug addicts have several problems, and several of them were well-known in the social welfare system or by the police. Half of the drug addicts had received treatment for their abuse and three persons were receiving treatment at the time of their death. A few--primarily among the youngest--had only used drugs for a short time while one third had abused drugs for several years. The greater part of the deaths were caused by heroin, which is found in different concentrations in the illegal market. Five of the dead persons had just been released from prison and three people had just left 24-hour care centres when they were found dead. Co operation between the various, treating authorities has to be given a high priority. PMID- 7483103 TI - [Richter hernia in trocar site after laparoscopic herniotomy]. AB - A case of an incarcerated Richter's hernia in a 12-mm trocar site is presented. A 72 year old man underwent laparoscopic herniorrhapy because of a recurrent inguinal hernia. On the sixth postoperative day he developed abdominal pain, nausea, vomiting and abdominal distension. Plain abdominal X-ray showed bowel obstruction. Computed tomography with oral contrast showed herniation of small bowel above the fascia. The patient was immediately reoperated, the intestine was reduced, and the fascial defect at the trocar site closed. Three days later he underwent surgery again due to a small perforation of the small bowel and a persistent fascial defect. The patient had an uneventful postoperative course. Herniation through a trocar site is a rare complication-incarceration extremely rare. We recommend that all fascial defects of 10 mm or more are closed sufficiently. PMID- 7483104 TI - [X-linked recessive bulbospinal neuropathy (Kennedy syndrome)]. AB - A case of X-linked recessive bulbospinal neuronopathy is reported. Neurophysiological findings were consistent with chronic partial denervation (motor axonopathy) and large fibre sensory axonopathy. DNA analysis showed an abnormal increased size of tandem CAG repeats. PMID- 7483105 TI - [Drug approval. Orientation from the drug department of the National Board of Health and Welfare and the scientific-ethical committee system]. PMID- 7483106 TI - [News on eclampsia]. PMID- 7483108 TI - [Reliability of adrenaline administered endotracheally]. PMID- 7483109 TI - [Death under anesthesia]. PMID- 7483107 TI - [Calcium channel blockers]. PMID- 7483110 TI - [Death under anesthesia. Definition, causes, risk factors and prevention]. AB - Death associated with anaesthetic procedures is rare, 1-4 deaths per 10,000 anaesthesias. However, each case gives rise to discussion about causality and who is to blame. Prospective studies are few, and comparison between them is difficult because of the use of different definitions of anaesthesia related death. A critically ill patient with impaired function of multiple organs seems to be at a higher risk of anaesthesia related death than a more healthy patient. However, no study has so far identified preoperative risk factors of anaesthesia related death. This is probably due to the low incidence of anaesthesia related death. The most common causes of anaesthesia related deaths are: 1) circulatory failure due to hypovolaemia in combination with overdosage of anaesthetic agents such as thiopentone, opioids, benzodiazepines or regional anaesthesia; 2) hypoxia and hypoventilation after for instance undetected oesophageal intubation, difficult intubation, technical failure in the anaesthetic equipment, or aspiration of gastric content, 3) anaphylactoid reactions including malignant hyperthermia, and 4) human negligence such as lack of vigilance or errors in the administration of drugs and in the maintenance and control of the anaesthetic equipment. We discuss the importance of continuing education for anaesthesiologists, development of a standard for surveillance during anaesthesia and quality control of the anaesthetic procedure with registration of undesired incidents. National registration of serious incidents will make it possible to determine the incidence of serious complications and death associated with anaesthesia. Hopefully this registration will provide information about causality and thereby facilitate prevention and improve patient safety during anaesthesia. PMID- 7483111 TI - [Mitochondrial encephalomyopathies]. PMID- 7483112 TI - [Problem-based medical teaching]. AB - A possible method of improving teaching at faculties of Health Sciences is the so called problem-based teaching method. A student is presented with a (clinical) medical case, which is to be investigated. By means of this method, the student himself will, under guidance, formulate his individual lack of knowledge, goals, priorities, methods, and evaluation. Educational observations show that the learning process is better when the problem-solving takes place among a group of students. A tutor acts as a catalyst for the process and gives feed-back through the problem-solving. He is therefore not an intermediary of knowledge to the group, but primarily refers the student to literature or knowledge experts at the faculty. International studies have not proven any unequivocal improvement in the students' ability to pass exams or to work as doctors, but both teachers and students seem to be more satisfied with the teaching method, which has stimulated the wish to study. The use at other faculties has not caused any large increase in expenditures. This should result in an increased use of problem-based teaching in medical education. PMID- 7483113 TI - [Iron status markers in pregnant women. No clinically significant connection between the values during the second trimester, later in pregnancy and after delivery]. AB - The aim of this randomized, double blind, placebo controlled parallel study was to evaluate whether iron status markers in pregnant women, measured early in the second trimester, could be used to predict levels later in pregnancy, and post partum. One hundred and twenty healthy pregnant women between 14 and 18 weeks of gestation were included. Sixty-three women were allocated to treatment with tablets containing 66 mg ferrous iron (as fumarate) daily, and 57 women to treatment with placebo. Haemoglobin, serum transferrin saturation, and serum ferritin were measured every fourth week during gestation, prior to delivery, one week and eight weeks post partum. Correlation matrices during pregnancy and post partum were calculated for each iron status marker, both in iron and placebo treated women. Haemoglobin, transferrin saturation and serum ferritin values at inclusion displayed steadily declining correlation coefficients with values obtained later in pregnancy. There were no clinically relevant correlations to values obtained eight weeks or less prior to delivery, or post partum. Serum ferritin values at inclusion could not be used to predict values later in pregnancy or post partum. Haemoglobin, transferrin saturation and serum ferritin values measured in the beginning of the second trimester appear unsuitable as guidelines for an individual iron prophylaxis during the remaining period of pregnancy. PMID- 7483114 TI - [Periorbital and orbital cellulitis in children]. AB - During a six-year period 28 febrile children with acute periorbital swelling were identified. Median age was 1.8 years (range, two weeks-nine years). When possible, the children were classified as having preseptal cellulitis (infectious focus in the subcutaneous tissue in front of the orbital septum) or postseptal cellulitis (focus in the paranasal sinuses). In six children a diagnosis of preseptal subcutaneous cellulitis was based upon the clinical presentation and positive culture results. Five children were additionally classified as having preseptal cellulitis due to severe conjunctivitis. Fifteen children were classified as having postseptal cellulitis due to infection in the paranasal sinuses (ethmoiditis). In seven of the latter cases the diagnosis was based upon the results of CT/MR-scans or operation. Two children could not be classified. All children were given antibiotic treatment and four had an operation. There were no recurrences or sequelae. In our study a presumed diagnosis could be made in retrospect in most cases, but at the time of presentation the clinical picture was often confusing. We found a CT- or MR-scan to be most helpful, especially in identifying the children requiring surgical treatment. PMID- 7483115 TI - [Percutaneous transthoracic needle aspiration biopsy. Diagnostic yield after negative fiberoptic bronchoscopy in patients with peripheral pulmonary infiltrations]. AB - The diagnostic potential of secondary transthoracic needle biopsy (TNB) following negative fiberoptic bronchoscopy in patients with peripheral circumscribed pulmonary lesions was evaluated in a retrospective study. The records from 224 patients who had TNB performed over a five-year period were reviewed. Of these, 103 patients met the criteria for inclusion in this series. The overall diagnostic yield of TNB in malignancy was 73.8% (54 of 73 patients). TNB allowed cytological classification of the tumour type in 72.2% (39 of 54 patients). Five of the 54 patients (9.3%) presented with small cell anaplastic bronchogenic carcinoma, diagnosed by TNB, and were treated with chemotherapy. Of the 49 patients with a negative TNB, 27 went on to diagnostic surgical procedures; 19 had malignancy, three benign tumour, two infection, and three sequelae after pulmonary infarction. The remaining 22 patients with undiagnosed lesions were followed long term, five showed progression of the pulmonary lesion suggesting malignancy. TNB appeared unsuitable in the diagnosis of benign lesions. Unspecific inflammation was not considered evidence of benignity, and therefore a definitive benign diagnosis was not made by TNB in this series. There were no serious complications to TNB. In 18.1% of the procedures a pneumothorax developed, indicating a chest tube in 8.6% of the procedures. TNB is a suitable diagnostic procedure with a high diagnostic yield in patients with peripheral, malignant pulmonary lesions. PMID- 7483116 TI - [Acute asthma outside hospital. The predictive value of pulse oximetry in manifestation of respiratory insufficiency]. AB - The objective of the study was to evaluate the predictive value of pulse oximetry before treatment of acute asthma in the prehospital setting. Unfavorable outcome was defined as need for mechanical ventilation during the acute attack. The study was carried out prospectively. The patients were divided into two groups: SpO2 < 80% and SpO2 > or = 80%. The study group consisted of 44 patients with 47 asthmatic attacks. Eleven patients had SpO2 < 80%, of whom two required mechanical ventilation. Thirty-three patients with 36 attacks had SpO2 > or = 80%, none required mechanical ventilation. Oximetry before treatment with a cut off point of < 80% had a specificity of 18% and a sensitivity of 100% for prediction of unfavourable outcome. We conclude at SpO2 < 80% in the acute asthmatic attack is a serious prognostic sign which should warn the physician that the patient's condition is lifethreatening. PMID- 7483117 TI - [Landfill gas]. AB - In a landfill mixture of gases, consisting principally of methane and carbon dioxide, may be produced by microbial degradation of organic waste under anaerobic conditions. Methane is explosive at concentrations between 5 and 15% by volume. Other gases, for instance hydrogensulphide, mercury and ethane, may be emitted at low concentrations, but usually do not represent a health hazard following normal atmospheric dilution. Indoor climate may be affected, though, in cases of accumulation in closed spaces. A case is presented where two persons died following an explosion caused by lighting a cigarette in their house which was surrounded on three sides by a landfill. The explosion occurred after heavy precipitation on a day with low atmospheric pressure. Methane measurements showed values consistent with risk of explosion. PMID- 7483118 TI - [Aortic dissection demonstrated by ultrasound scanning]. AB - The clinical features of aortic dissection may be nonspecific and confusing. Three patients with different symptomatology diagnosed primarily by ultrasound are reported. In all cases the dissection was confirmed by dynamic CT and arteriography. The sonographic characteristics of aortic dissection are briefly described. PMID- 7483119 TI - [Dose calculations in radioiodine therapy of benign thyroid diseases]. PMID- 7483120 TI - [Halothane and obese patients]. PMID- 7483121 TI - [Liver resection for metastases]. PMID- 7483122 TI - [Stethoscopy of carotid artery--for what purpose?]. PMID- 7483123 TI - The epidemiology of erectile dysfunction. AB - During the last two decades, significant advances have been made in the understanding of male sexual dysfunction. Concomitantly, a marked increase in both clinical and research activity in the field of male erectile dysfunction has led to a better evaluation and more treatment options. The prevalence and incidence are dependent on the definitions used, the diagnostic tolls, and the treatment options. Using standard definitions as suggested by the NIH Consensus Conference and improving our diagnostic and treatment options will have a major impact on the epidemiology of ED. A summary of the risk factors for ED is presented in Table 3. Still more epidemiologic research is essential to further understand the distribution as well as the prevalence of ED in certain ethnic groups, chronic conditions, and as a result of surgery and trauma. These studies will help us improve our diagnostic skills as well as our therapeutic options. PMID- 7483124 TI - Sexuality and aging. AB - Because aging is frequently associated with medical conditions likely to impair sexual performance, there is a tendency to view sexual changes in older men as the result of pathology, overlooking the effect of natural processes of aging in sexuality. This article discusses the sexual responses in the aging male, the physiologic aspects of aging male sexuality, psychologic aspects of aging and sexuality, studies on aging male sexuality, a psychobiological investigation of healthy aging men, medical causes of erectile dysfunction in elderly men, and the effect of medications on the sexual functioning of aging men. PMID- 7483125 TI - The penis as a vascular organ. The importance of corporal smooth muscle tone in the control of erection. AB - The intrinsic biological complexity of penile erection and the multifaceted nature of erectile dysfunction are just beginning to be fully appreciated. This article describes how mechanistic studies of the local control of penile erection, with specific emphasis on the primary role of the corporal smooth muscle, contribute to the improved understanding, diagnosis, and treatment of erectile dysfunction. PMID- 7483126 TI - Neural control of penile erection. AB - Filling of the sinusoidal spaces with blood due to smooth muscle relaxation results from parasympathetic neural pathway activation and probably simultaneous inhibition of sympathetic outflow. The final common pathway for proerectile fibers is represented by the cavernous nerves and fibers controlling detumescence and flaccidity originating in the sympathetic chain. The hypogastric nerve could represent an accessory proerectile pathway unmasked by a sacral spinal cord lesion. Nitric oxide, which can be colocalized with VIP and acetylcholine, is the main proerectile neurotransmitter and noradrenaline is considered to be the major antierectile agent. Reflexive erection elicited by recruitment of penile afferents involves both autonomic and somatic efferents. This reflex is mediated at the spinal cord level and modulated by supraspinal influences. Serotonergic pathways originating in the raphe nuclei may mediate inhibitory control on reflexive erections. The hypothalamic medial preoptic area is an important integrating center and dopamine may regulate penile erection at this level. Neuroendocrine regulation may vary depending on the context in which erection occurs, for example, coitus, in response to extrinsic or psychogenic stimuli, and rapid eye movement sleep. PMID- 7483127 TI - Psychological issues in diagnosis and treatment of erectile disorders. AB - Psychological issues are salient in every case of erectile dysfunction because sexual function is a psychosomatic process. The best overall assessment is made through separate psychosocial interviews of patient and partner. Interview technique requires comfort and a nonjudgmental attitude. A combination of psychological and physiological treatments offer the best outcome for many cases. PMID- 7483128 TI - Use of nocturnal penile tumescence and rigidity in the evaluation of male erectile dysfunction. AB - The role of nocturnal penile tumescence monitoring in helping to distinguish psychogenic from organic impotence has been the subject of research for several decades. Tumescence monitoring alone, while it does provide useful information, imposes limitations on the diagnostic inferences that can be drawn concerning the adequacy of erectile function. This led to the development of nocturnal penile tumescence and rigidity monitoring (NPTR). Although there is no single test that enables physicians to diagnose the precise etiology and degree of impotence, NPTR provides useful information in a rather noninvasive and inexpensive manner allowing one to direct patients to the appropriate treatment. PMID- 7483129 TI - Diagnosis and management of endocrine disorders of erectile dysfunction. AB - Organic causes of erectile dysfunction with androgen deficiency may be associated with aging, systemic illness, and a number of specific endocrine disorders stemming from pituitary, thyroid, and adrenal dysfunction. Central hypogonadism is the main mechanism in the majority. Erectile dysfunction in diabetes mellitus is caused by chronic complications due to poor metabolic control. Diagnosis and management of these disorders are discussed, as is the need for tight glycemic control in men with diabetes. PMID- 7483130 TI - Investigation of erectile dysfunction. Diagnostic testing for vascular factors in erectile dysfunction. AB - To date, several accurate tests for diagnosing vascular ED may be chosen. It is necessary to be well aware of the purpose of testing: global assessment of erectile capacity in preparation for auto-injection therapy, or detailed assessment of arterial and erectile response in preparation for surgical treatment. Pharmacotesting may be sufficient for the majority of patients. Other, more invasive tests are reserved for preparing surgical treatment or scientific studies. PMID- 7483131 TI - Penile revascularization. AB - Penile revascularization surgery to correct penile vascular insufficiency has undergone several modifications since its introduction. Further improvement in the surgical success rate will be achieved with improved understanding of erectile physiology, diagnostic methodology, and both microsurgical and radiological techniques. PMID- 7483132 TI - Intracavernous injection therapy for male erectile dysfunction. AB - Intracavernosal injection therapy has consistently produced erections in 60% to 70% of patients in whom it is tried. About 70% of patients enter long-term therapy and there is a dropout rate of 20% to 50% within the first year. The most effective current regimen is a mixture of papaverine-phentolamine and prostaglandin E1, that also seems to have a relatively low incidence of priapism and fibrous nodule formation. Patient education is vitally important to the success of a program. Patient satisfaction rates are high. PMID- 7483133 TI - Long-term results of penile prosthetic implants. AB - This article is intended to provide the implanting surgeon with information that can be used as a guideline for proper informed consent for the patient who is to have a penile prosthesis. The modern penile prostheses are quite reliable with an approximate 5% mechanical failure rate for the inflatable prosthesis after being in place for 5 years. Although approximately 90% of patients will have a functioning device at 5 years, only 70% will be completely satisfied with the device and this satisfaction rate is about the same for the partners. Late infection is possible and the use of prophylactic antibiotics during the time that infection risk might be possible is recommended. PMID- 7483134 TI - Use of penile prosthetic implants in patients with penile fibrosis. AB - For a subset group of impotent patients with penile fibrosis, the implantation of a penile prosthesis is their only therapy for restoration of erectile function. In many cases, using a penile prosthesis requires additional corporal reconstructive procedures to rebuild fibrotic corporal bodies. Unfortunately, the surgeon is rarely forewarned that reconstructive surgery will be necessary despite a variety of historic factors that would suggest the possibility of encountering unsuccessful corporal dilatation. To be forewarned is to possibly decide that implant surgery is contraindicated, thereby denying some patients one last opportunity to have potency restored. Corporal reconstruction with or without use of synthetic graft material will not be suitable for all patients in whom severe corporal fibrosis is encountered. There is a relatively high risk of infection in these patients and an indepth, preoperative discussion with those patients in whom corporal reconstruction is considered a possibility is essential in order to be assured of the patient's motivation to "go the extra mile." For some patients, as an elective procedure, there is definitive unwillingness to undergo such a complicated procedure. In those cases, it is usually mutually agreed that if extensive corporal fibrosis is encountered, the procedure will simply be abandoned and no attempt will be made to carry out corporal reconstruction. Many patients with penile fibrosis are young which attests to their greater motivation to not permit their sex life from being terminated at an unacceptably early age. Quite rightly, they maintain that benefits of corporal reconstruction, when successful, far outweigh the risks. PMID- 7483135 TI - A practical approach to the evaluation and treatment of erectile dysfunction. A private practitioner's viewpoint. AB - There are many changes occurring in the delivery of health care that are economically impacting our urology practices. Certainly, the days of being primarily prostate and TURP doctors are over. Incorporating an impotence component in our practices is an effective method of filling the void left by treating BPH. Treating impotence will not require any significant additional equipment or staff. Not only are there opportunities within our own practices, there are thousands of men suffering from impotence in our communities who need our help and our expertise. Finally, using the two-visit technique, most men can be evaluated and begin a treatment program that will result in a successful resolution to their sexual problem in a cost-effective fashion. PMID- 7483136 TI - Oral and topical treatment of erectile dysfunction. Present and future. AB - A great deal of progress has been made in the pharmacological treatment of erectile dysfunction. At present, however, the most effective therapies require intracavernosal injections with a number of associated drawbacks. An increasing number of oral and transdermal agents have been introduced clinically or are at various phases in their development. It is evident that severe end-organ disease probably will not result in successful systemic therapy. Nevertheless, in men with intact or mildly dysfunctional erectile mechanisms, noninvasive treatments can offer some measure of success. Further study of individual and synergistic activity of available compounds is underway. PMID- 7483137 TI - Augmentation phalloplasty. AB - Aesthetic procedures can increase the girth and visual length of the penis. Dermal-fat grafts increase penile circumference without the complications that result from fat injections. Release of the suspensory ligaments with skin advancement may increase flaccid penile length. Suprapubic lipectomy and Z-plasty of a penoscrotal web enhance penile appearance. Accurate diagnosis and meticulous technique are mandatory. PMID- 7483138 TI - Prognostic factors for bladder cancer. AB - Cancer of the urinary bladder is the fifth most common cancer in men and the second most urological malignancy in Western society [17], with an incidence rate per year of 29.8/100,000 males. Bladder tumors are distinguished as either invasive or superficial: invasive tumors are generally associated with poor prognosis, while 20-30% of superficial carcinomas recur and progress to become invasive and metastic [26, 27]. The most common prognostic factors for classification of urothelial cancer are staging and grading, which are based on morphological criteria. In the past decade, however, other criteria have been developed as a possible prognostic aid to better disease management, such as expression of specific cell surface antigens, DNA content, chromosomal aberrations, gene rearrangements and point mutations [26, 7]. Since most tumors of the bladder are carcinomas and are associated with dedifferentiation and high metastatic capability, we investigated whether reduced expression of so-called differentiation factors in combination with increased cell motility might be correlated with tumor progression. PMID- 7483139 TI - Establishment and characterization of a new cell line from human bladder cancer (JMSU1). AB - A new human bladder cancer cell line designated JMSU1 has been established from malignant ascitic fluid of a 75-year-old Japanese man with bladder cancer, and maintained in culture for more than 7 years and over 240 passages. Inverted phase contrast microscopy revealed that JMSU1 was composed of morphologically distinct cells (polygonal to spindle-shaped cells), showing morphological heterogeneity in vitro. Histological examination of xenografts showed poorly differentiated transitional cell carcinoma, resembling the original tumor. Immunohistochemical staining for cytokeratin and electron microscopic examination suggested that JMSU1 was of epithelial origin. Chromosome analysis gave a modal number of 69 with no Y chromosome. Isozyme analysis (LDH, G6PD, and NP) showed the mobility pattern of human type B. DNA fingerprint analysis demonstrated that there was no cross-culture contamination of JMSU1 during the passages. In conclusion, a newly established and well-characterized cell line, JMSU1, offers promising material for the investigation of the biological properties of bladder cancer. PMID- 7483140 TI - Photodynamic efficiency of liposome-administered tetramethyl hematoporphyrin in two human bladder cancer cell lines. AB - The main problems presented by superficial bladder carcinoma, its high recurrence rate and multifocal appearance, require treatment of the bladder as a whole. Photodynamic therapy (PDT) is one such experimental treatment for superficial bladder carcinoma, involving the administration of a photosensitizer that accumulates in the tumor tissue, and subsequent irradiation of the tumor with light. Since the photosensitizers used in PDT suffer from several drawbacks, new photosensitizers are being sought. Drug delivery systems are also being investigated for the administration of hydrophobic photosensitizers and enhancement of photodynamic efficiency and tumor selectivity. In this study we examined a new photosensitizer, tetramethyl hematoporphyrin (TMHP), in two human bladder cancer cell lines. In the first pair of the experiments, TMHP was bound to unilamellar liposomes. Cellular uptake, dark toxicity and photodynamic efficiency were then studied. Fluorescence microscopy showed TMHP localization in the cytoplasm in a perinuclear region, sparing the nucleus. Dark toxicity occurred after incubation of cells with TMHP above a concentration of 20 micrograms/ml. Irradiation was carried out using an argon-pumped dye laser emitting a wavelength of 630 nm at a fluence of 3.6 and 7.2 J/cm2. Before irradiation, cells were incubated with TMHP at concentrations of 2.5 and 5 micrograms/ml for 1 h. Cell survival rates after incubation with 5 micrograms/ml TMHP and irradiation at 7.2 J/cm2 were 15.7% of control cells for Rec and 4.5% for Waf cells. Uptake studies showed a higher intracellular TMHP concentration in Waf than in Rec cells. This correlates with the higher PDT efficiency seen in Waf cells.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7483141 TI - Effects and systemic uptake of the new mitomycin C analogue KW-2149 in beagle dogs after intravesical administration. AB - The present study was designed to evaluate the local effects of the new mitomycin C analogue KW-2149 after intravesical instillation, together with its penetration into the systemic circulation in healthy beagle dogs. Two reference dogs were treated with two instillations of mitomycin C (30 mg in 30 ml phosphate buffer). Four dogs were given two, three, four and six instillations, respectively, of KW 2149 (60 mg in 30 ml phosphate buffer). KW-2149 concentrations measured in the systemic circulation were very low and were frequently found to be below the limit of determination. The number of instillations had no influence on the KW 2149 concentrations measured in the systemic circulation. Blood analysis showed no systemic toxicity. The histopathological findings in the bladder were comparable in both groups. The number of instillations had no influence on the severity of the lesions found in the bladder wall. On the basis of its in vitro activity KW-2149 can be regarded as a promising agent for intravesical treatment of superficial bladder cancer. PMID- 7483142 TI - Biomodulation of 5-Fu cytotoxicity by folinic acid and its stereoisomers: in vitro experiments with different cell lines of prostatic cancer. AB - The results of cytotoxic chemotherapy for advanced, hormone-escaped prostate cancer have been disappointing. Evaluation of the effect of new drugs or new combinations with already known ones is required. The antimetabolite 5 fluorouracil (5-FU) has been shown to be active in prostate cancer, acting via inhibition of thymidylate synthase, an essential enzyme in DNA de novo synthesis. Experiments with cell lines of different tumors have shown that 5-FU activity can be modulated by addition of the coenzyme tetrahydrofolic acid (folinic acid). We investigated the effect of folinic acid and its stereoisomers on 5-FU action in different cell lines of prostate cancer. It was found that addition of non-toxic folinic acid led to a two- to fourfold better antiproliferative effect of 5-FU. The unnatural 6R isomer, which is a compound of chemically synthesized folinic acid, inhibited the modulatory effect of the natural 6S isomer. Our results indicated that a combination of folinic acid and 5-FU may result in a better response of patients with hormone-resistant prostate cancer than of patients treated with 5-FU alone. PMID- 7483143 TI - Doxorubicin treatment of rabbit renal VX-2 carcinoma: nephrotoxicity, serum parameters and weight. AB - Serum electrolytes, creatinine, urea, protein, albumin, bilirubin and glucose were examined every 4 days until time of death in rabbits with VX-2 carcinoma implanted in one kidney. The rabbits were treated with doxorubicin, nephrectomy or combinations thereof and observed for up to 1 year. Rabbits treated with doxorubicin only showed a slight creatinine rise initially, but over time creatinine reached almost the same concentration as that in nephrectomized rabbits receiving equivalent doses of doxorubicin. Creatinine concentrations increased significantly above the normal range following nephrectomy combined with doxorubicin. Doxorubicin nephrotoxicity in rabbits occurs at lower doses than previously reported. In all rabbits the parameters except creatinine remained stable within the established normal ranges, except for the last 4 days before time of death in the animals with metastatic disease. Weight loss was the best parameter for making a prognosis for an individual rabbit, since peak weight was noted 16-20 days before death. In experimental work with VX-2 carcinoma, weight is thus the most important indicator of the time at which rabbits not responding to treatment can be put to death to avoid unnecessary suffering before the end of the experiment. PMID- 7483144 TI - Expression of cell adhesion molecules in an established and characterized new human renal cell cancer line, CCF-RC7. AB - In order to investigate the importance of cell adhesion molecules (CAMs) in renal cell carcinoma (RCC), a cell line, designated as CCF-RC7, was established from a human RCC of the clear cell type. CCF-RC7 was passaged over 50 times in vitro for 3 1/2 years. The cell line has an epithelial morphology and a doubling time of 30 h, forming colonies in soft agar with an average efficiency of 10.4% and producing clear cell tumors in athymic nude mice. CCF-RC7 cells have an aneuploid hypotetraploid karyotype with a modal chromosome number of 82 and rearrangements in chromosomes 9, 12 and 14. Immunohistochemical and flow immunocytometric analyses revealed high expression of ICAM-1 (CD54), and Hermes antigen (CD44), which was significantly upregulated by cytokine and PMA treatment. VLA-4 was expressed on approximately 20% of tumor cells and could not be altered by cytokine or PMA stimulation. High expression of sialyl Lewis X was also demonstrated by immunohistological examination. This newly characterized cell line will serve as a useful model for the study of CAMs during hematogenous metastasis and host defense mechanisms in human RCC. PMID- 7483145 TI - Gammopathy associated with advanced prostate carcinoma. AB - As proteinuria was found to be common in patients with prostate cancer, the possible presence of elevated urinary immunoglobulin (Ig) levels was investigated. First morning urine samples from 30 patients with androgen dependent (AD) and 43 patients with androgen-independent (AI) prostate cancer were tested. A sensitive, solid-phase radioimmunoassay and polyclonal antibody interacting with IgG, IgA and IgM was used to screen urine samples. Compared with 15 normal, age-matched, healthy subjects, urinary Ig levels were elevated in 10 of 30 (33%) patients with AD prostate cancer and in 24 of 43 (56%) patients with AI prostate cancer. In the latter group, five of seven (71%) patients with prostatic SCC had elevated urinary Igs. Further analysis of ten urine samples containing increased urinary Ig levels, using antibodies specific for each heavy chain (gamma, alpha and mu) as well as each light chain (kappa and lambda), indicated that patients with a high or moderate elevation in Ig levels had polyclonal gammopathy, whereas those with a low increase in urinary Igs had monoclonal gammopathy of the IgG class. These results indicate, for the first time, a high frequency of an abnormal increase in Ig levels in patients with advanced prostatic carcinoma. PMID- 7483147 TI - Serous-lined extramural ileal valve as a new continent cutaneous urinary outlet: an experimental study in dogs. AB - A new technique for the construction of a cutaneous continent catheterizable outlet suitable for urinary reservoirs is presented. The technique entails the creation of an ileal reservoir outlet by implantation of a tapered ileal segment into a serous-lined extramural tunnel. The procedure was carried out in eight experimental dogs, and the results were compared with a control group of five dogs for which a nontunnelled tapered ileal outlet draining a similar reservoir was constructed. Clinical, radiological and urodynamic evaluation provided evidence that this new outlet is easily catheterizable. Its continence mechanism is more reliable than that of a simple tapered outlet. PMID- 7483146 TI - Nitroxergic innervation of the human ureterovesical junction. AB - Nitric oxide synthase (NOS) immunohistochemistry and nicotinamide adenine dinucleotide phosphate diaphorase (NADPH-d) histochemistry were used to investigate the distribution of nitroxergic, i.e., nitric oxide-synthesizing, neuronal perikarya and processes in the human ureterovesical junction (UVJ). Tissue specimens obtained from two cadaver kidney donors and two patients undergoing radical cystectomy for bladder cancer were examined. Clusters of NOS immunoreactive neurons were localized in extramural ureterovesical ganglia. NOS containing nerve fibers traveled within large extramural nerve trunks and marched among smooth muscle bundles. Extramural and intramural blood vessels were encircled by varicose NOS-positive axonal processes. The distribution of NOS immunoreactivity paralleled the staining pattern for NADPH-d activity. Urothelium stained strongly for NADPH-d activity but showed no NOS immunolabeling. Specimens from all four patients investigated showed similar staining patterns. Our results suggest that nitric oxide, a potent smooth-muscle-relaxing neurotransmitter in the autonomic nervous system, plays a physiologic role in opening the human UVJ. PMID- 7483148 TI - A study of recurrent stone formers with special reference to renal tubular acidosis. AB - Forty-five patients with recurrent renal stone were examined for distal renal tubular acidosis (dRTA) defects by acid challenge test (150 mg ammonium chloride/kg body weight). Their 24-h urine samples were analysed for creatinine, calcium, oxalic acid, inorganic phosphorus, uric acid, magnesium and citric acid. One-hour urine samples before acid load and hourly samples for the 7 h following acid challenge test were collected and analysed for creatinine, calcium, citric acid, inorganic phosphorus, titratable acidity, and ammonium. The incidence of distal RTA defect was 22.2% in the patients examined. The major biochemical characteristics in RTA patients compared with patients without RTA were: (a) significantly higher urinary pH, (b) significantly lower excretion of citric acid, (c) no significant difference in calcium excretion and (d) a tendency toward lower titratable acidity and ammonium excretion. PMID- 7483150 TI - [Urinary incontinence]. PMID- 7483149 TI - [Advances in the diagnosis and therapy of prostate cancer. International symposium, 13 March 1993, Vienna]. PMID- 7483151 TI - [Advanced prostatic carcinoma--which hormone therapy when?]. PMID- 7483152 TI - [Introduction to the statistical aspects of planning clinical oncologic phase III studies]. AB - A phase III clinical trial is a comparative study in which one assesses the relative efficacy of a treatment or treatments. It generally has one of the following objectives: 1. To determine the effectiveness of a new treatment relative to the natural history of the disease, for example when comparing a new treatment to not treatment or to placebo in an adjuvant setting. 2. To determine if a new treatment is more effective than the best current standard therapy (at the risk of increasing the toxicity). 3. To determine if a new treatment is as effective as the best current standard therapy but is associated with less severe toxicity or a better quality of life (equivalence trial). Clinical trials must be properly designed in order to answer such questions with a high degree of certainty. The purpose of this paper is to present several concepts which must be taken into account during the process of designing a clinical trial. PMID- 7483153 TI - [Clinical protocols in treatment of prostatic carcinoma in Europe. The role of EORTC and other organizations]. AB - In the last few years the EORTC has conducted multiple clinical studies on the therapy of prostate cancer. In most cases with incidental unifocal prostate cancer, no therapy is necessary, as the disease is rarely progressive or the cause of death. Current EORTC studies are evaluating the timing of hormone therapy, as it is still unknown whether therapy should be started at the time of diagnosis or when the patient becomes symptomatic. Different hormone therapies are useful for locally progressive prostate cancer and as effective than radiotherapy in the case of progression and survival. The preliminary results of a current study on total androgen blockade for 3 months before radical prostatectomy show a positive effect for some of the patients with a T2 carcinoma. Inconclusive results in studies on total androgen blockade versus orchiectomy in metastasising prostate cancer have lead to a meta-analysis of 22 studies by the EORTC. This only showed a trend towards better survival time and longer interval till progression with total androgen blockade but the group of patients with small volume metastatic disease benefit greatly from this treatment. The importance of estrogen therapy and the treatment with high dose non steroidal antiandrogens as well as the use of intermittent hormonal therapy in progressive disease will have to be assessed in future studies. Chemotherapy is not very effective due to its low response rate and its short lived effect. PMID- 7483154 TI - [Recent multicenter study protocols in the USA for patients with metastatic prostatic carcinoma]. AB - Combined androgen blockade for metastatic prostate cancer has become the standard against which on-going clinical trials are measured. Benefit of combined androgen blockade with an LHRH agonist (leuprolide) and an antiandrogen (flutamide) has been noted, particularly for those with minimal disease. This has led to timely accrual of patients to a trial testing whether orchidectomy with flutamide is superior to orchidectomy alone. Positive findings with permanent and intermittent withdrawal of flutamide has prompted the design and implementation of additional trials. Suramin has been reintroduced in trials of chemohormonal intervention. Past success with combined androgen blockade in lengthening survival and improving quality of life for patients with metastatic prostate cancer has prompted enthusiasm for broadening the base of clinical studies for this disease. PMID- 7483155 TI - [Theoretical considerations and initial clinical results of intermittent hormone treatment of patients with advanced prostatic carcinoma]. AB - Androgen suppression is the routine approach to the treatment of advanced prostate cancer. Using intermittent androgen suppression by taking the advantage of the reversible action of medical castration results in the maintenance of apoptotic potential. The experiments in the androgen-dependent androgen-dependent Shionogi carcinoma tumor model as well as clinical experience in a group of men with prostate malignancy are presented in this report. These consecutive cycles of androgen withdrawal and replacement afford an improved quality of life when the patient is off therapy. It is possible to reduce toxicity, cost of treatment and to delay tumor progression. Whether survival is affected in a beneficial or adverse way still remains to be studied. PMID- 7483156 TI - [Renaissance of estrogen therapy in advanced prostate carcinoma?]. AB - Oestrogen therapy or orchidectomy reigned supreme for the treatment of metastatic prostatic cancer for almost 40 years. In the last 15 years many alternative agents have been tested alone and more recently in combination. This contribution evaluates the results of some of these newer therapies considering relief of symptoms, toxicity, quality of life, survival, and benefit to the community. The results are compared with those of oestrogen therapy and with the way in which such treatment might be amended in light of more modern knowledge. PMID- 7483157 TI - [Intramuscular depot estrogens (Estradurin) in treatment of patients with prostate carcinoma. Historical aspects, mechanism of action, results and current clinical status]. AB - More than 50 years ago, orally given estrogen was already used in the treatment of prostate cancer. Due to cardiovascular side-effects with a high morbidity of 25%, this treatment has not become standard. Recent investigations show that parenteral application reduces the risk of cardiovascular side-effects, because it avoids the first passage through the liver with high concentrations of estrogen which normally occur after oral application. Therefore, an increased synthesis of so-called "steroid-sensitive" liver proteins, such as coagulation factors (especially factor VII) can be avoided. This newer parenteral estrogen application shows encouraging results of a cheap and effective hormonal therapy with a low rate of side-effects in patients with prostate cancer. PMID- 7483158 TI - [High energy focused ultrasound in treatment of benign prostatic hyperplasia]. AB - Transurethral resection of the prostate (TURP) and open adenectomy are regarded the "golden standard" in the treatment of benign prostate hyperplasia (BPH). Various alternative treatment forms (microwaves, laser, radio frequency, focused ultrasound) have been introduced recently. They are all aimed at reducing the morbidity related to TURP while achieving comparable efficacy at the same time. Since December 1992, 35 patients with BPH have been treated with high-intensity focused ultrasound (HIFU-P) in our department. By 6 weeks after HIFU-P, mean Qmax had improved from 6.5 ml/s to 12.1 ml/s. Residual volume dropped from 207 ml to 95 ml, IPSS from 20.1 to 9.7. Both subjective as well as objective improvements remained stable after 3 and 6 months. Urinary tract infections were observed in 3 patients, macrohemato-spermia in all, and macrohematuria (caused by the suprapubic catheter) requiring blood transfusion in 1 patient. PMID- 7483159 TI - [MESA (microsurgical epididymal sperm aspiration) and IVF (in vitro fertilization). A therapy concept in treatment of male infertility]. AB - Microsurgical epididymal sperm aspiration (MESA), or sperm microaspiration retrieval technique (SMART), in conjunction with in vitro fertilization is a successful therapy in male infertility. From November 1991 to March 1994 a total of 29 attempts at MESA with subsequent IVF were made. Of 48 aspirations, 37 were successful and 13 attempts at IVF were possible, 6 of which were successful with 10 subsequent embryo transfers. In all, 3 pregnancies were achieved and 1 boy was born. In conclusion, microsurgical spermaspiration in conjunction with in vitro fertilization is a way of treating male infertility with a chance of achieving paternity with the partner's own sperm; the chances are probably better with intracytoplasmatic sperm injection. PMID- 7483160 TI - [Preoperative assessment of tumor aggressiveness in localized prostatic carcinoma]. AB - In 106 consecutive patients with localized prostate cancer digital rectal examination (DRE), preoperative prostate-specific antigen (PSA) determination and results of systematic sextant biopsies (TRUS 6Bx) of the prostate were analyzed for their value in the estimation of the aggressivity of tumors. In all patients with negative pelvic lymph nodes radical retropubic prostatectomy was performed. Tumor aggressiveness was defined as capsular penetration (pT2 versus pT3) or positive surgical margins in patients with pT3 tumors. Neither DRE nor preoperative PSA level was helpful in predicting capsular penetration or positive surgical margins. However, the number of positive core biopsies and the identification of Gleason 4 or 5 tumors within positive biopsy specimens correlated with capsular penetration and positive surgical margins. These results can be used to create a score, based on DRE, PSA, TRUS 6Bx, and Gleason 4 or 5, that might be helpful in predicting tumour aggressivity in patients with localized prostate cancer. PMID- 7483161 TI - [Is radical prostatectomy a suitable model for determination of PSA half-life?]. AB - A review of the literature relating to PSA half-life reveals great variability in absolute values and pharmacokinetic models. A critical view is needed, however, since some authors suggest that the PSA half-life has implications for diagnosis and prognosis after radical prostatectomy. The aim of our study, therefore, was to characterize the value of PSA half-life determination after radical prostatectomy. Serial serum PSA detections were performed in 16 patients with localized prostatic cancer who had undergone radical prostatectomy. Serum PSA was detected on days 0, 1, 2, 3, 6, 9, 12, 15, 18, after radical prostatectomy. In all patients elimination of PSA from serum followed a biphasic logarithmic decay pattern indicating a two-compartment model of first order elimination kinetics (t1 = 1.01 +/- 0.06 days, t2 = 3.42 +/- 0.23 days; P < 0.00001). In this two compartment model 56.3 +/- 4.8% of the preoperative PSA serum concentration was cleared by the first compartment. To find a biological correlative for the first compartment a mathematical model was developed to approximate the effect of operative blood and plasma loss on PSA serum concentration. In this model changes of hematocrit were used to estimate blood and plasma loss. These calculations showed that 50.12 +/- 3.04% of the preoperative PSA serum concentration was excreted by operative blood loss. This value was not significantly different from the clearance rate calculated for the first compartment. It is, therefore, concluded that the determination of PSA half-life after radical prostatectomy without correction of the operation-related PSA loss is only of limited value. PMID- 7483162 TI - [Testicular biopsy in azoospermia before fertilization interventions--uni- or bilateral? Initial results of a prospective study]. AB - Azoospermia can have testicular or posttesticular causes. For confirmation of the diagnosis, a testicular biopsy is recommended. Whether bilateral biopsy is necessary is the subject of some controversy. In a prospective study, 44 consecutive male patients (mean age 32.4 [20-52] years) with azoospermia were examined in our andrological department. Spermatogenesis was analyzed by means of a semi-thin technique and with reference to a modified Johnsen-Score. There was no difference in median size between the right (13.5 [8.5-15] ml) and the left testis (11 [7-15] ml). In the analysis of the score values, we found no differences between the two testes (median scores: right side 3 [2-7.2], left side 3.75 [2.1-7.45]). In one quarter of the patients who had a difference of more than 2 ml in testicular size the higher score was found in the smaller testis. In linear regression analysis, no correlation was found between differences in testicular volume and score differences. There was a qualitative difference in spermatogenesis in two patients. In no patient was a carcinoma in situ detected. The above results led us to the conclusion that a unilateral biopsy is adequate from both qualitative and quantitative aspects. PMID- 7483163 TI - [Paraneoplastic syndromes]. PMID- 7483164 TI - A brief history of mentoring. PMID- 7483166 TI - Fibroblast growth factor signalling and myogenic development. PMID- 7483165 TI - Gene therapy to restore prostacyclin presence to injured endothelium. AB - These preliminary studies demonstrate the feasibility of restoration of prostacyclin synthesis in mechanically-injured porcine carotid arteries following angioplasty. Our initial data suggest the possibility of inhibiting thrombus development by adenovirus-CMV-PGHS-1 therapy in the initial 10 days following angioplasty. PMID- 7483167 TI - Neurogenetics: now and the future. PMID- 7483168 TI - Neuroimmunophysiology of the gastrointestinal mucosa: implications for inflammatory diseases. AB - In conclusion, studies of the neuroimmunophysiology of the intestinal mucosa of the past 5-8 years have demonstrated an important role for the immune system in modulating water and electrolyte transport as well as intestinal motility in the gut. Activation of mast cells and phagocytes leads to heightened Cl- and water secretion, as well as changes in intestinal motility which leads to diarrheal states. These diarrheal responses are self-protective; they rid the intestine of offending microorganisms and antigens. Our investigation of this response has uncovered a new immune accessory cell Cz, the intestinal myofibroblast. This cell seems to play an important role in amplifying the immune signal. This cell is probably also important for the secretion of growth factors onto the epithelium and also the secretion of collagen which results in fibrosis under diseased states. These intestinal myofibroblasts are prolific prostaglandin producers, an important finding because prostaglandin synthesis inhibition has been shown to decrease the development of neoplasia in the gut. Thus, these intestinal myofibroblasts may have other important roles in addition to just modulating water and electrolyte secretion or gut motility. Our laboratory is now engaged in studying these intestinal myofibroblasts in some detail hoping to better understand the biology of these interesting cells. PMID- 7483169 TI - Unique organ specific characteristics of amino acid metabolism in early development. PMID- 7483170 TI - Differentiated thyroid cancer long-term impact of initial therapy. PMID- 7483171 TI - The Jeremiah Metzger Lecture. Academic medicine: building on the strengths of the past as we approach a cloudy future. PMID- 7483172 TI - Strategies to achieve balance in the physician workforce. PMID- 7483174 TI - The saga of an academic clinician: from micropuncturist to macroadministrator. PMID- 7483173 TI - An overview of internal medicine manpower: impact of the increased need for primary care physicians. PMID- 7483175 TI - Career tracks of Kaiser scholars in general medicine. PMID- 7483176 TI - Polycythemia vera--a climatological disease. PMID- 7483177 TI - Fungal infections complicating orthotopic liver transplantation. PMID- 7483178 TI - Winner of the Theodore E. Woodward Clinical Award. Monster of the Chesapeake Bay- Vibrio vulnificus. PMID- 7483179 TI - Astrovirus gastroenteritis. PMID- 7483180 TI - Obesity-related hypertension and the insulin resistance syndrome. PMID- 7483181 TI - Asymptomatic cardiac ischemia pilot (ACIP). AB - This pilot study demonstrated that (1) patients with CAD and asymptomatic cardiac ischemia can be randomized to medical or revascularization strategies using a complex and demanding protocol, (2) asymptomatic cardiac ischemia can be suppressed in 40-50% of patients with clinically advanced coronary disease with relatively low to moderate doses of medication titrated over a period of 12 weeks (3). Revascularization was the most effective of the treatment strategies studied in reducing ischemia. Any type of therapy, whether it be drugs or revascularization requiring repetitive monitoring with ambulatory ECG or other methods to detect ischemia over a long period of time, will escalate the cost of quality medical care for our patients. Thus, the health care costs implications and treatment of asymptomatic ischemia are enormous. But the apparent cost advantage of treating only symptoms, that is ignoring all ischemia, could disappear if treatment of ischemia reduces the risk of adverse events. The clinical question to be addressed in the future is what is necessary to reduce the cardiac-event rates of death and myocardial infarction in this group of patients? Will more aggressive drug therapy eliminate more ischemia and will therapy directed at the elimination of all detectable ischemia improved clinical outcome better than therapy directed to control angina only? These questions can only be answered by a large clinical trial. The results of such a trial will provide the basis and rationale for safe and effective therapy for patients with coronary disease and evidence of cardiac ischemia. Whatever the answer to this important medical and scientific question is, it will have tremendous economic implications.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7483183 TI - [Viewpoints]. PMID- 7483184 TI - ["Medical research confidentiality" could promote research and protect personal rights--conquering data protection related research deficits in Germany]. AB - The workgroup of the German Medical Societies (AWMF) has asked for the introduction of a law ruling "Medical Research Secrets". The aim is to reduce the obstacles put on important medical research, especially in epidemiology, mainly caused by the legal protection of privacy of personal data. This aim could be reached by a legal protection of medical research data in adjunction to the legal status of the professional secrecy at present effective for physicians. The realization of the legal protection of specific research projects could be approved by local ethic-committees, they would bind the individual researchers to a closely protected secrecy and so enable the transfer of personal data from different sources without the obstacles of the present legal protection of privacy data and the legal professional secrecy effective for physicians. This is of importance for the combination of research support for the gain of future patients and protection of personal privacy for patients of today and would mean a correction of an erroneous German development of a legally perfectionized mistrust of physicians involved in research. It will be shown, that cancer registers are necessary, but that present laws ruling cancer registers are hostile to research and will depreciate the research location Germany. PMID- 7483185 TI - [Proper management of numbers--some references for the clinically active physician]. AB - Even common medical practice cannot be performed without using figures and some kind of basic statistics. Unfortunately this field is complicated by many pitfalls. In the paper several examples are given how to find and how to avoid them. This is completed by a selection of underlying principles of statistics. PMID- 7483182 TI - The Gordon Wilson Lecture. In vivo gene therapy: a strategy to use human genes as therapeutics. PMID- 7483186 TI - [Aging and cancer. An analysis of cancer mortality in Germany 1900 to 1990]. AB - The unification of Germany rendered possible the comprehensive overall calculation and subsequent analysis of cancer mortality of an entire population of 60 to 80 million people over a period of 90 years. 1. The total mortality rates (all forms) in the various age groups are largely constant. The increase in absolute terms is a consequence of the changing age distribution in the German population resulting in a larger number of people in higher age groups. 2. The age-related constancy of total cancer mortality can be confirmed statistically via longitudinal and cross-sectional approaches focussing on changing subject groups per calendar year as well as in a diagonal manner using the same group of subjects per year of birth. 3. Each age group has its own characteristic and exponentially-at higher age-increase cancer mortality rate which is independent of influences by e.g. personal environment or "generation". 4. The constancy of total cancer mortality implies a change in the distribution of organ manifestations which equilibrates any increases or decreases. Available data on organ manifestation of cancer from the year 1950 on are the basis of this observation. 5. Age-related constancy of total cancer mortality is a basic principle of cancer epidemiology. PMID- 7483187 TI - [Prognostic relevance of oncologic after-care]. AB - Follow-up programs in oncology after potentially curative therapy intent to detect recurrent cancer, to initiate a new curative therapy, to detect side effects and secondary cancer. The prognostic relevance of follow-up in oncology can be defined as the rate of asymptomatic detected potentially curable recurrences. Follow-up programs of different tumors usually combine various diagnostic methods in a standardized schedule. The prognostic relevance of this follow-up programs is investigated in only very few studies. While efficacy of the follow-up in gastro-intestinal cancer must be questioned, follow-up in non seminomatous germ cell cancer has proven to be effective. This shows that global statements about prognostic relevance of follow-up can not be made. It is therefore necessary to investigate the actual follow-up programs in clinical studies whether they are effective to improve the prognosis in cancer patients. Where the actual programs fail to be effective new risk-adapted follow-up concepts must be created. New approaches in diagnosis and treatment of cancer have to tested whether they can improve the effect of follow-up programs. PMID- 7483189 TI - [Claim review in occupational disability insurance]. AB - In a deregulated market characterised by increasing competitiveness and improved conditions, care in assessing proposals in additional disability benefits will take on an even more significant role. The assessment has to be based on overall analysis of the moral and objective risk. This entails risk assessment of financial, professional, economic and medical factors, and should also include experience gained from the specific area of benefits involved. In terms of getting closer to our clients, the possibilities of limiting risk should be offered to an increasing extent on an individual, transparent and differentiated basis. Despite every effort to exercise caution, the time factor, i.e. the speed required to issue policies, should not be forgotten. PMID- 7483188 TI - [MEDRISK--an expert system for medical risk assessment]. AB - The Munich Reinsurance Company has developed a rule-based expert system for assessing substandard risk in life, disability and accidental death benefit. It is one of the most comprehensive medical expert systems yet conceived and currently includes entries for over 7500 impairment terms. Based on the most up to-date insurance medical knowledge MEDRISK allows underwriters, irrespective of their level of experience, to process both simple and highly complex cases. The system which takes account of the interactive effect that can exist between different impairments as well as the influence which occupational factors can exert, always produces consistent and case-specific decisions. The number of impairments and types of insurance included in MEDRISK can be expanded. After tests at Munich Re and at a number of insurance companies, the system ist now ready to be launched in German speaking markets. PMID- 7483190 TI - [Excess mortality of severely handicapped patients in mandatory health insurance]. AB - The introduction of compulsory long term care insurance as of 1.1. 1995 has temporarily brought to an end a whole range of controversial discussions, which have been strongly coloured by party political interests. The originally planned expenditure figure of at least DM 30 billion represents an enormous outlay in these time of economic recession. It is already abundantly clear that the premium payments will not be sufficient in the long run, nor will it be possible to increase these premiums arbitrarily. This therefore elicits the question as to whether it is possible to calculate in advance the cost of treatment for those "in serious need of nursing care" on the basis of various factors such as age, sex, underlying illness, therapy and social integration. Up to now, there has been very little statistical analysis of these parameters. For this reason, the company MDK has carried out a preliminary survey (which so far has only looked at a limited number of cases) in order to obtain a general idea of the variations in the period required for long term care. This survey has shown that there are two main groups of cases requiring care, which can be differently assessed on the basis of age and sex. On the one hand, there are those in serious need of nursing care, who, due to a life-threatening disease or as a result of an acute deterioration of an existing chronic condition (e.g. severe KHK, cirrhosis of the liver, decompensated renal insufficiency) die a relatively short time after the application for care is made.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7483191 TI - [A method of forming a rhinostoma in case of the stenosed lumen of the lacrimal sac]. AB - A new method for the formation of a rhinostoma in patients with stenosed lumen of the lacrimal sac is proposed. It consists in the following: the incision of the soft tissues and the opening in the bone are made as low as possible to be closer to the lacrimal sac, and the nasolacrimal duct is used for the formation of a rhinostoma. Seventy-two operations were performed in patients with stenosed lumen of the lacrimal sac: 32 using sparing dacryocystorhinostomy (group 1) and 40 using the proposed technique (group 2). The results of 66 operations (30 in group 1 and 36 in group 2) were observed in remote periods (from 8 months to 9 years). In group 1 complete effect was attained in 20 out of 30 cases (66.7%), in group 2 in 33 out of 36 (91.6%). Hence, the proposed method for the formation of rhinostoma is highly effective. PMID- 7483192 TI - [Results of the treatment of retinoblastoma]. AB - The treatment efficacy is analyzed in 262 patients (335 eyes). Besides traditional enucleation, organ-sparing methods were used in combination with polychemotherapy with or without external irradiation. The authors emphasize the advisability of polychemotherapy supplementing the therapeutic complex in retinoblastoma. PMID- 7483193 TI - [Possibilities of the use of low-energy laser effects on the ciliary body in optic nystagmus]. AB - The accommodation potential of the eye is markedly reduced in patients with optic nystagmus. Trans-scleral exposure of the ciliary body to infrared low-energy laser at wavelength 1.3 microns may be used to repair the accommodation potential of the eyes and improve the visual function in nystagmus. Laser stimulation of the ciliary zone was carried out in 62 patients with optic nystagmus. As a result, the accommodation potential was 1.417 times increased and the closest point of clear vision approached the eye by 1.309 diopters. Far vision acuity increased by 1.334 times, close one by 1.258 times. A stable positive result persisted for 3 to 6 months, after which laser stimulation may be repeated, or other means of repairing the accommodation system of the eye be resorted to, such as training after Avetisov-Mats or exercises with a home ergographer. PMID- 7483194 TI - [Laser irradiation: study of total effect of irradiation of the eyeball. A clinical study]. AB - Lipid peroxidation products, antioxidant system parameters, somatotrophic hormone, corticosteroids, fibronectin, and cyclic nucleotides were measured in the blood of 119 patients with open-angle glaucoma and mature senile cataract before and after a course of exposure of the eyeball to He-Ne laser. Statistically significant changes in the examined parameters were revealed, explaining to a great measure the antiinflammatory and vasodilating effects of laser and its ability to intensify the repair processes. PMID- 7483195 TI - [Results of the use of vasoactive preparation pikamilon in pigmented retinal abiotrophy]. AB - Eighty patients with pigmented retinal abiotrophy (PRA) and 20 controls were examined. The perfusion pressure and arteriovenous coefficient are markedly reduced in patients with stages 1-2 PRA, in comparison with controls. Deterioration of the visual function in patients with stages 3-4 PRA vs. that in stages 1-2 was associated with a more marked reduction of the hemodynamic parameters and the status of ocular vessels. Pikamilon therapy in a dose of 2 ml of 10% solution once a day intramuscularly for 10 days led to improvement of the visual function and ocular hemodynamics in patients with PRA. Treatment efficacy was higher in patients with disease stages 1-2. Pikamilon is recommended for the treatment of patients with PRA. PMID- 7483196 TI - [Effects of tobacco smoking on ocular thermoregulation]. AB - Time course of eyeball temperature in response to smoking of a cigarette was studied in healthy smoking male volunteers aged 19 to 28 (24 men, 48 eyes). Before smoking the mean temperature in the limbic area of the eye was 31.7 +/- 0.1 degree C (the reference age-specific value is 32 +/- 0.02 degree C). It was reliably reduced 15 and 30 min after smoking (p < 0.01), recovering only in 90 min after smoking. Hence, tobacco smoking impairs the trophics of ocular tissues, these disturbances may be referred to signs of aging or to the initial stages of many well-known pathologic shifts in the eye associated with tobacco smoking (optic nerve atrophy, retinal degeneration, grave course of glaucoma, etc.). PMID- 7483198 TI - [Improvement of the method of prehospital protection of vision]. AB - The authors describe an improved method for the prevention of ocular diseases in children, consisting in formation of risk groups starting from female consulting centers ("provisional" group), then at maternity hospitals ("provisional" and "reliable" groups), and then at the outpatient clinic after examination by an ophthalmologist ("actual" and "reliable" groups). This work does not require extra staff or finances, and it helps reduce the incidence of ocular diseases, poor vision, and blindness almost twofold. PMID- 7483197 TI - [Spectral filters as a method of therapeutic correction]. AB - Spectral filters were used for additional correction of vision in 67 patients: in 15 adults with initial cataracts (intensive yellow filter), 26 children with albinism (yellow-brown filter), 14 children with macular hypoplasia (orange filter), and 12 children with aphakia after removal of congenital cataracts (yellow filter). Selection of the filter density is carried out using a special method including visocontrastometry, examination of sensitivity to lateral light, and study of color perception thresholds. Use of filters resulted in improvement of the vision acuity by 43.5% in patients with initial cataracts, by 10% in those with albinism, by 20% in those with macular hypoplasia, and by 22% in those with aphakia; moreover, an improvement of the frequency-contrast characteristics was observed, as well as a reduction of photophobia, and a reduction of vision amplitude in patients with nystagmus. The possible applications of spectral correction are discussed. PMID- 7483199 TI - [Visual evoked potentials in the evaluation of visual acuity in infants]. AB - Vision acuity was assessed by visual evoked potentials (VEP) in response to low frequency chess pattern reversion in 31 full-term healthy babies aged 6 to 28 weeks. The mean vision acuity of babies aged 6 to 9 weeks was 0.5 +/- 0.01, of those aged 16 to 18 weeks 0.1 +/- 0.01; in babies aged 24 to 26 weeks vision acuity was at least 0.14, but still lower than in normal-sighted adults. The authors emphasize the advantages of low-frequency chess pattern reversion in assessment of vision acuity from VEP in babies. PMID- 7483200 TI - [Experience in laser treatment of primary glaucoma]. AB - An Interdistrict Laser Center has been functioning in Vladivostok for 10 years. Laser exposure was used in the treatment of 749 patients with primary glaucoma. Yatagan-1, Orion-3001, YAG Pegaus-3002 lasers (Germany) were employed. In 55% cases primary glaucoma was concomitant with essential hypertension, in 12% with diabetes mellitus. Laser iridectomy, laser goniopuncture, laser trabeculoplasty were used. Hypotensive effect lasting for at least 6 months was attained in 69.1% cases. Complications presenting as hemorrhages in the anterior chamber, which spontaneously resolved on day 2 after exposure, were observed in only 4% cases. Laser therapy in patients aged 50 to 70 is the only means of reducing ophthalmic tone and preserving the visual function because of somatic diseases present at this age. Laser treatment of primary glaucoma brings about an appreciable economic effect and creates favorable conditions for subsequent microsurgery. PMID- 7483201 TI - [Experience in the undergraduate training of ophthalmologists]. AB - The authors present their experience gained in the training of ophthalmologists at the Chair of Ocular Diseases of the Krasnoyarsk Medical Institute. In 1991 the interns, instead of traditional state examinations, maintained their diploma research, which was assessed as their skills and knowledge in the field of ocular diseases, social hygiene, and public health organization. The authors prove that such diploma research is an integral part of undergraduate training and one of the types of individual work at higher educational institutions, and enumerate the topics of diploma research. PMID- 7483202 TI - [Bioactive substances in the regulation of vitreoretinal proliferation]. PMID- 7483203 TI - [Principal factors affecting postoperative astigmatism after cataract extraction]. PMID- 7483204 TI - [From the history of ophthalmological services in the Vyatka region]. PMID- 7483205 TI - [Intraocular metallic crystalline lens for correction of aphakia]. AB - Technological characteristics of a new in principle intraocular lens (IOL) consisting of a leukosaphire lens and elastic tantalum haptics are presented. The advantages of metallic crystal IOLs are validated: complete chemical and biological inertness, longevity, high transparency, and easy implantation in the capsular sac. Surgical recommendations are offered and highly positive results of clinical trials analyzed (more than 600 implantations followed up for up to 4 years). Metallic crystal IOLs for intracapsular correction of aphakia developed at the S. I. Vavilov All-Russian Research Center and St. Petersburg Division of the Moscow Research and Technological Complex "Ocular Microsurgery" and manufactured by the IOL Joint Stock Company are recommended for wide clinical use. PMID- 7483206 TI - [Interlamellar circular keratoplasty for correction of high myopia]. AB - A new method is proposed for the correction of high myopia: Interlamellar circular keratoplasty. This method has been used in 15 patients (17 eyes) aged 18 to 54 with 9 to 17 diopters myopia. Vision acuity attained was at least 0.5 in 10 (60.3%) patients; in 7 (35.2%) patients vision acuity was 0.3 without correction, that is, was equal to the maximal vision acuity with the optimal correction. The highest refraction effect was 15.0 diopters. The patients were followed up for 3 months to 16 years. The proposed method for the correction of high myopia is highly effective, simple, and is recommended for clinical practice. PMID- 7483207 TI - Pathologic features of naturally occurring juvenile polyarteritis in beagle dogs. AB - Eighteen young Beagle dogs (eight males and 10 females), ages 6-40 months, with canine juvenile polyarteritis syndrome (CJPS), a naturally occurring vasculitis and perivasculitis of unknown etiology, were necropsied, and their tissues were examined by histopathologic and histochemical methods. The condition is characterized by recurring episodes of an acute onset of fever (> 40 C) and neck pain that persist for 3-7 days. The major histopathologic alterations were a systemic vasculitis and perivasculitis. During the febrile, painful period of CJPS, the vascular lesions ranged from a histiocytic-lymphocytic periarterial infiltration to transmural arterial inflammation with concomitant fibrinoid necrosis and vascular thrombosis. Massive periarterial accumulations of inflammatory cells were common and often extended into adjacent tissues. The small- to medium-sized muscular arteries of the heart, cranial mediastinum, and cervical spinal meninges were consistently involved. Vasculitis occasionally occurred in other organ systems. The vascular lesions in dogs examined during clinically normal periods consisted of intimal and medial fibrosis, ruptured elastic laminae, and mild perivasculitis; these lesions were probably related to previous episodes of vasculitis. Eight dogs that had experienced repeated acute episodes also developed splenic, hepatic, and renal amyloidosis. The clinical signs, laboratory abnormalities, and the vascular lesions suggest that the condition may be immune-system mediated. CJPS may serve as a naturally occurring animal model of human immune-system-mediated vasculitides such as polyarteritis nodosa, infantile polyarteritis, and Kawasaki disease. PMID- 7483208 TI - Clinical and clinical laboratory correlates in sea otters dying unexpectedly in rehabilitation centers following the Exxon Valdez oil spill. AB - Following the Exxon Valdez oil spill, 347 oiled sea otters (Enhydra lutris) were treated in rehabilitation centers. Of these, 116 died, 94 within 10 days of presentation. Clinical records of 21 otters dying during the first 10 days of rehabilitation were reviewed to define the laboratory abnormalities and clinical syndromes associated with these unexpected deaths. The most common terminal syndrome was shock characterized by hypothermia, lethargy, and often hemorrhagic diarrhea. In heavily and moderately oiled otters, shock developed within 48 hours of initial presentation, whereas in lightly oiled otters shock generally occurred during the second week of captivity. Accompanying laboratory abnormalities included leukopenia with increased numbers of immature neutrophils (degenerative left shift), lymphopenia, anemia, azotemia (primarily prerenal), hyperkalemia, hypoproteinemia/hypoalbuminemia, elevations of serum transaminases, and hypoglycemia. Shock associated with hemorrhagic diarrhea probably occurred either as a direct primary effect of oiling or as an indirect effect secondary to confinement and handling in the rehabilitation centers. Lightly oiled otters were less likely to die from shock than were heavily oiled otters (22% vs. 72%, respectively). Heavily oiled otters developed shock more rapidly and had greater numbers of laboratory abnormalities, suggesting that exposure to oil was an important contributing factor. PMID- 7483209 TI - A new inherited muscular disorder in Japanese quails (Coturnix coturnix japonica). AB - Thirteen adult mutant (LWC strain) Japanese quails (Coturnix coturnix japonica), between the ages of 8 and 60 weeks were examined for a progressive muscular disorder. The disorder, inherited as an autosomal dominant trait, was clinically apparent as early as 28 days of age; it was characterized by generalized myotonia, muscle stiffness, and muscle weakness. Affected birds were identified by their inability to lift their wings vertically upward and by their inability to right themselves when placed on their dorsum. Electromyographic studies in two mutant quails showed high-frequency repetitive discharges comparable to those of myotonic runs. These discharges persisted after nerve resection. The distinctive histopathologic changes in the various muscles examined were ring fibers, sarcoplasmic masses, and internal migration of sarcolemmal nuclei. A slight decrease in the size of type IIB muscle fibers and a slight increase in the size of type IIA fibers were observed in the M. pectoralis thoracicus of affected quails. In older affected birds, inter- and intrafascicular fatty infiltration with replacement of type IIB fibers by fat cells was seen in the pectoral muscles. Single fiber necrosis, nonspecific lymphorrages, and variations in the muscle fiber size and shape were also noted. The typical muscle lesions and multisystem involvement, which was manifested by testicular degeneration and atrophy in the male LWC specimens and bilateral lenticular cataracts in 6 of 13 affected mutant quails, suggest resemblance of this new inherited muscular disorder to myotonic dystrophy in man. PMID- 7483210 TI - Pathogenesis of porcine reproductive and respiratory syndrome virus infection in gnotobiotic pigs. AB - The pathogenesis of porcine reproductive and respiratory syndrome virus (PRRSV) was determined in gnotobiotic pigs by studying the sequential development of microscopic lesions and sites of virus distribution and replication. Thirty-two pigs (three pigs/infected group and one pig/control group) were inoculated by nasal instillation of either PRRSV isolate ATCC VR-2332 (total dose 10(2.6) TCID50) or uninfected cell culture supernatant. Infected and control pigs were euthanized at 12 hours, and 1, 2, 3, 5, 7, 14, and 21 days postexposure (PE). Gnotobiotic pigs experimentally infected with PRRSV were viremic by 12 hours PE and subsequently developed pneumonia, lymphadenopathy, vasculitis, myocarditis and encephalitis. Lung lesions developed by day 3 PE, persisted through day 21 PE and were characterized by alveolar septa thickened by macrophages, alveolar proteinaceous and karyorrhectic debris, alveolar syncytial cells, and multifocal type II pneumocyte hypertrophy. Lymph node lesions varied in distribution and severity and were characterized by germinal center hypertrophy and hyperplasia, lymphocyte necrosis, multiple cystic spaces, and polykaryocytes within the cystic spaces. Heart lesions were a late feature of infection and all infected pigs had heart lesions on day 21 PE characterized by subendocardial, myocardial, and perivascular foci of lymphocytes. Vasculitis also varied in distribution and severity and affected all sizes of vessels. Results of this experiment indicate that PRRSV is a multisystem disease characterized initially by viremia with subsequent virus distribution and replication in multiple organs causing interstitial pneumonia, vasculitis, lymphadenopathy, myocarditis, and encephalitis. PMID- 7483211 TI - Antibodies to canine granulocyte colony-stimulating factor induce persistent neutropenia. AB - A severe persistent neutropenia developed in a rabbit that was injected intradermally with 120, 60, 60, and 120 micrograms of recombinant canine granulocyte colony-stimulating factor (cG-CSF) on days 1, 22, 31, and 44, respectively. The neutropenia was present from day 44 to day 205. The nadir of the neutropenia (60 cells/microliters) occurred in conjunction with peak antibody titer (640,000) to cG-CSF on day 58. The immune antiserum from this rabbit reacted positively for cG-CSF on Western blot analysis. The immune antiserum also neutralized the activity of cG-CSF. On day 160, examination of the bone marrow showed marked granulocytic hypoplasia and mild erythroid hyperplasia. On day 205, the rabbit was still neutropenic (430 cells/microliters), even though the last injection of cG-CSF was given 161 previously. Necropsy on day 205 showed that there was still mild granulocytic hypoplasia with mild erythroid hyperplasia. Because of the lack of any inflammatory foci found at necropsy and the granulocytic hypoplasia, it was thought that the neutropenia was most likely due to decreased production and was not a consumptive process. It is hypothesized that the antibody that was produced to cG-CSF neutralized the effect of endogenous rabbit granulocyte colony-stimulating factor and prevented the normal proliferation and maturation of the rabbit neutrophils. PMID- 7483212 TI - Preneoplastic and neoplastic lesions of rat hereditary renal cell tumors express markers of proximal and distal nephron. AB - Long-Evans (Eker) rats carry a mutation that predisposes them to develop spontaneous renal cell tumors of two morphologic patterns: solid chromophilic masses or cystic lesions lined by eosinophilic cells. Previous studies have suggested that these tumors arise from the proximal tubules. In the present study, lectin-binding characteristics and cytokeratin expression of various stages of hereditary rat renal epithelial neoplasia were examined to localize the portion of the nephron from which tumors arise. Lectin-binding histochemistry has been used as a marker of cell surface glycoprotein expression, thought to be important in the differentiation of benign from malignant epithelial lesions and in the determination of their cell of origin. The presence or absence of keratin intermediate filaments in the rat nephron has been used to identify nephron segments. The polyclonal antibody to high- and low-molecular-weight cytokeratin stained the cells of the collecting ducts but not the proximal or distal tubules. Binding to the proximal tubules by the lectins Conavalia ensiformis (Con A), Dolichas biflorus, Ricinus communis (RCA-1), and Triticum vulgare and to the distal tubules by Con A, RCA-1, Arachis hypogaea (PNA) with and without neuraminidase, and the antibody for cytokeratins was demonstrated. The lectin binding and cytokeratin staining patterns of rat hereditary renal cell carcinoma, adenoma and the preneoplastic lesions of atypical tubules and hyperplasias suggest that cystic adenomas arise from the distal nephron, principally the collecting duct, whereas the solid atypical tubules, hyperplasias, and adenomas arise from the proximal nephron, principally the proximal tubule. PMID- 7483213 TI - Canine bullous pemphigoid (BP): identification of the 180-kd canine BP antigen by circulating autoantibodies. AB - Human bullous pemphigoid (BP) is an immune-mediated blistering disease characterized by autoantibodies against BP antigens (230/180 kd), which are constitutive glycoproteins of hemidesmosomes found in basal keratinocytes. Blistering diseases similar to human BP have been reported in dogs. IgG deposits at the basement membrane zone (BMZ) are a common feature of canine BP. Although circulating anti-BMZ IgG autoantibodies have been demonstrated in some cases of canine BP, the specific skin protein targeted by these autoantibodies has not been identified. In this study, we characterized the antigenic target of the autoantibodies in the serum from a 3-year-old castrated male Pit Bull Terrier with BP. Direct immunofluorescence of the patient's skin demonstrated IgG deposits at the dermal-epidermal junction. Indirect immunofluorescence demonstrated autoantibodies in the patient's serum that stained the epidermal roof of salt-split canine skin and left the dermal floor unstained. These serum autoantibodies did not stain normal intact dog skin but labeled intact bovine tongue. Direct immunoelectron microscopy of the dog's skin revealed IgG deposits within the hemidesmosomes of the basal keratinocytes. Western immunoblotting experiments showed that canine keratinocytes express both the 230-kd and 180-kd bullous pemphigoid antigens, and the autoantibodies from the patient's serum recognized the 180-kd bullous pemphigoid antigen in proteins extracted from canine and human keratinocytes. Canine BP has many parallel features with human BP including similar immune deposition of IgG within hemidesmosomes and a hemidesmosome-associated 180-kd glycoprotein target for circulating autoantibodies. PMID- 7483216 TI - Senile plaques and other senile changes in the brain of an aged American black bear. AB - A female American black bear (Euarctos ursus americanus) over 20 years old had shown epileptiform neurologic signs starting in March 1992 and was found dead unexpectedly 8 months later. At necropsy, pulmonary and intrabronchial hemorrhage was noted. In the brain, the leptomeninges exhibited slight thickening, and petechiae were evident in the hippocampus. Histopathologic examination of the brain revealed several senile changes: numerous senile plaques, amyloid deposition in cerebromeningeal arterioles, mineral deposition in the pallidum, and numerous corpora amylacea in the cerebellum. PMID- 7483214 TI - Toxic effects of butylated triphenyl phosphate-based hydraulic fluid and tricresyl phosphate in female F344 rats. AB - Triaryl phosphates, including tricresyl phosphate (TCP and butylated triphenyl phosphates (BTP), are used in the commercial manufacture of plastics, lubricants, and hydraulic fluids. Recent reports implicate these compounds as endocrine and reproductive toxicants that can cause cholesteryl lipidosis in adrenocortical (AC) and ovarian interstitial (OI) cells, suggesting altered metabolism of steroid hormones or cholesterol or of both. We investigated potential mechanisms of BTP and TCP toxicity to determine if there were functional abnormalities of the adrenal cortex or ovary. Groups of intact (nine or 12) and ovariectomized (six) female F344 rats, 10-12 weeks of age, received 0, 0.4 g/kg TCP, or 1.7 g/kg BTP in sesame oil vehicle or 1.7 g/kg neat BTP for 20, 40, or 60 days. All rats administered BTP and TCP developed cholesteryl lipidosis in AC and OI cells; the TCP-treated group was most severely affected. Serum concentrations of androstenedione and corticosterone were unchanged, but estradiol levels were significantly (P < or = 0.05) elevated in BTP- and TCP-treated groups (14.5 times and 37.5 times greater than controls, respectively). Vaginal cytology revealed that BTP- but not TCP-treated females had abnormal reproductive cycles that were significantly prolonged in diestrus (3 times greater than control). There were significant elevations in serum total cholesterol (TCP-treated group was 1.3 times greater than controls), low-density lipoprotein (TCP-treated group was 1.8 times greater than controls), alanine transaminase (BTP-treated group was 2 times greater than controls), and albumin (a major serum estradiol-binding protein; BTP treated group was 4.6 g/dl vs. 3.6 g/dl for controls).(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7483215 TI - Endothelial MHC class II antigen expression and endarteritis associated with Marek's disease virus infection in chickens. AB - Experimental Marek's disease virus (MDV) infection in chickens was used to study the early pathogenesis of virus-induced atherosclerosis. Previous investigations using this model have reported the occurrence of atherosclerotic lesions after approximately 7 months postinfection. In this study, a total of 75 susceptible Cornell P-line chickens were inoculated intraperitoneally with the CU-2 strain of MDV at 3 days of age and subsequently perfused for histologic examination. At 2, 4, 8, 13, and 20 weeks postinoculation, the ascending aorta and the brachiocephalic and coronary arteries were evaluated for early changes. Expression of class II major histocompatibility complex (Ia) antigen by the vascular endothelium was demonstrated by indirect immunodetection as early as 2 weeks after virus inoculation. This change was followed by significant thickening of the intimal layer associated with mononuclear cell infiltration. All the arteries examined from the MDV-infected chickens were affected. Preliminary immunohistochemical staining showed the presence of CD3+ CD4+, and CD8+ cells among the infiltrating cells. The results suggest that an immunopathologic mechanism may be involved in the early pathogenesis of MDV-induced atherosclerosis in chickens. PMID- 7483217 TI - Biliary cystadenoma of cats. AB - Published surveys of feline neoplasia have not specifically included biliary cystadenoma, and there is only one case report in the literature. This report is a compilation of 13 feline cases and provides a description of clinical, pathologic, immunohistochemical, and ultrastructural aspects of biliary cystadenoma as well as a discussion of comparative pathology of biliary cystadenoma in human beings and speculative histogenesis. PMID- 7483219 TI - Primary gastrointestinal lymphosarcoma with epitheliotropism in three Shar-pei and one boxer dog. AB - Three male Shar-pei dogs and one female spayed Boxer dog ranging in age from 4 years, 4 months to 7 years, 8 months had 4-7-week histories of diarrhea, progressive anorexia, weight loss, and vomiting. The dogs were depressed, lethargic, and hypoalbuminemic, with losses of 23-39% of body weight. Endoscopic examination revealed gastric ulceration with areas of mucosal hyperemia and pallor. Small nodules with occasional central depressions were present in three dogs. Gastric and proximal duodenal mucosal biopsies showed neoplastic lymphocytes infiltrating the lamina propria, with invasion of the superficial and glandular epithelial basement membrane. Immunohistochemical evaluation of two dogs was negative for anti-canine IgG and positive for anti-CD3. Necropsy of the Boxer and two Shar-peis confirmed epitheliotropic gastric and small intestinal lymphoma in all three dogs, with mesenteric lymph node and hepatic involvement in two and one dog, respectively. PMID- 7483218 TI - Spontaneous renal tubular carcinoma in Fischer-344 rat littermates. AB - Two of 632 Fischer-344 rats in a food restriction study had spontaneous, bilateral, multicentric renal tubular cell carcinomas. Although there were 104 litters represented in this study, both rats that developed this rare neoplasm were from the same litter. The littermates, one male and one female, were in the food-restricted treatment groups (60% of ad libitum intake) and were 550 and 447 days old, respectively, at death. The probability that the two rare bilateral renal neoplasms occurred by a chance event in littermates is approximately 0.8%. The apparent familial predisposition for development of specific types of neoplasms emphasizes the importance of randomization of individuals into treatment groups and consideration of lineage for rare tumors. PMID- 7483221 TI - Epithelioma of the fourth ventricle in a goat. AB - We describe an epithelial neoplasm arising from the fourth ventricle of a goat and extending into the subjacent cerebellum along the spaces of Virchow-Robin. The neoplastic cells are consistent with squamous epithelium based upon light microscopic morphology and cytokeratin immunoreactivity. The lack of overt keratin formation and the lack of an exophytic component distinguishes this neoplasm from intracranial epidermoid cysts, resulting in the classification of this caprine tumor as an epithelioma. PMID- 7483222 TI - Disseminated adenovirus infection associated with cutaneous and visceral hemorrhages in a nursing piglet. AB - An unusual adenovirus infection in a conventionally raised nursing pig was characterized clinically by hemorrhagic diathesis of the skin and microscopically by disseminated viral inclusions in the endothelial, interstitial, and epithelial cells of the skin, kidney, spleen, liver, heart, and small and large intestines. This disease was confirmed ultrastructurally by demonstration of typical adenoviral particles in the nuclei of endothelial cells in the myocardium and of interstitial cells in the small intestine. This is the first report of disseminated adenovirus infection in a nursing pig in North America. PMID- 7483220 TI - Hepatopathy associated with excessive hepatic copper in a Siamese cat. AB - A 2-year-old spayed female Siamese cat was presented with clinical liver disease characterized by anorexia; depression; elevations in serum levels of alanine aminotransferase, aspartate aminotransferase, and lactate dehydrogenase; hyperbilirubinemia; and icterus. Liver biopsy diagnosed hepatocellular degeneration with marked centrilobular hepatocellular accumulation of rhodanine positive brown granules. Subsequent postmortem examination revealed similar granular material in the epithelium of the proximal convoluted tubules and collecting ducts of the kidney and alveolar epithelium and macrophages in the lung. The liver and kidney copper concentrations were 4,074 and 792 ppm dry weight, respectively. Hepatic degeneration in this cat apparently was due to excessive accumulation of copper. PMID- 7483223 TI - Extramedullary hematopoiesis in the choroid plexus of five dogs. AB - Five dogs euthanatized because of refractory seizures were found to have hematopoietic elements in the interstitium of the choroid plexus at the level of the fourth ventricle. None of the dogs had significant hematologic or cerebrospinal fluid abnormalities. The extramedullary hematopoiesis was confined to the central nervous system and consisted of megakaryocytes, immature granulocytes, and rubricytes in two dogs and of one predominant cell population in each of the other three dogs. These findings are unique, and factors possibly contributing to the formation of a hematopoietic inductive microenvironment in the choroid plexus are cytokine-neurokine homologies, locally altered vascular supply, and aberrant functioning of bone marrow-derived central nervous system macrophages. PMID- 7483225 TI - Recovery of hepatic function and latent mortalities in sheep exposed to the blue green alga Microcystis aeruginosa. AB - Seventeen sheep died, and many others showed signs of hepatogenous photosensitivity after being exposed to Microcystis aeruginosa at Lake Mokoan, Victoria, Australia. Two groups of sheep were observed, and their hepatic recovery was monitored by means of serum biochemical tests during the subsequent six months. During the first three weeks, their serum gamma-glutamyl transferase activity and bilirubin concentration declined rapidly to normal levels, and the signs of hepatogenous photosensitivity disappeared. Serum bile acid concentrations were above normal for almost three months. Thirty-four per cent of the sheep died during the observation period, and the serum biochemical tests provided no clear indication of the cause of these delayed mortalities. This study suggests that sublethal exposure to microcystins may cause prolonged morbidity and delayed mortality. PMID- 7483224 TI - Fibrosarcomas in two rats arising from hepatic cysts of Cysticercus fasciolaris. AB - Two of five male Sprague-Dawley rats with hepatic tapeworm cysts developed large multinodular fibrosarcomas. Fibrosarcomas envelope tapeworm cysts, invaded the serosa of multiple organs, and extended through the diaphragm into the pleural cavity. Light microscopy, immunohistochemistry, and electron microscopy supported the diagnosis of fibrosarcoma. The parasites were identified as Cysticercus fasciolaris, the larval stage of Taenia taeniaeformis. The development of sarcomas in rats induced by Taenia sp. is thought to be attributable to the chronic inflammatory reaction of the capsule. There are parallels between these and other tumors occurring in mice and cats with suggested chronic inflammatory etiologies. PMID- 7483226 TI - Production of four identical calves by the separation of blastomeres from an in vitro derived four-cell embryo. AB - The blastomeres of two in vitro derived four-cell embryos were separated and transferred individually into surrogate zonae pellucidae, then co-cultured with bovine oviductal epithelial cells for five days until blastulation. Pairs of the quarter blastocysts were co-transferred with trophoblastic vesicles into each of four synchronised Holstein heifers, three of which were diagnosed pregnant at 28 days gestation, carrying twin fetuses. Four genetically identical bull calves were delivered by elective caesarean section at term pregnancy. One pregnancy was terminated at 56 days. PMID- 7483227 TI - Assessing the economic impact of immunisation against East Coast fever: a case study in coast province, Kenya. AB - The cost of immunising cattle against East Coast fever by the infection and treatment method has been calculated for a pilot scheme in Kaloleni Division of the Coast Province of Kenya by using a spreadsheet model. The cost was calculated to be KSh 544 (US$25) per animal (in 1990 values). If a farmer were to bear all this cost, immunisation would be financially profitable in grade cattle, but the benefits of immunisation would not be sufficient to justify the immunisation of zebu cattle. For these animals, the cost of immunisation would have to be in the range of KSh 230 to KSh 415 per animal, or the farm-gate price of milk would have to increase by at least 80 per cent from KSh 7.50 to 13.50/litre, or the government would have to subsidise the cost either partially or fully. The first two possibilities are realistic, because the costs of routine immunisation are likely to be lower than for the pilot scheme, and because the increasing demand for milk is likely to push up prices in the liberalised markets. If both the grade and zebu cattle in Kaloleni Division were targets for immunisation, it is estimated that there would be 14,500 head for immunisation annually, costing an estimated KSh 8 million. The spreadsheet model used to assess the economics of immunisation in the Kaloleni Division could be applied to determine the government or private veterinary service charges for immunisation that would be financially profitable to farmers in a defined cattle production system in any division, district or country. The model could also be used to estimate the annual total number of cattle for immunisation in a target cattle production system and thus help with the financial planning for the exercise. PMID- 7483228 TI - Allopurinol in the treatment of canine visceral leishmaniasis. PMID- 7483229 TI - AgNOR count as a marker in ethmoid carcinoma in bovids. PMID- 7483230 TI - Live animal exports. PMID- 7483231 TI - Detection of FeLV antigen. PMID- 7483232 TI - Weight:length ratios in chelonia. PMID- 7483233 TI - Spotlight on E coli O157. PMID- 7483234 TI - Local and plasma antibody responses to the parasitic larval stages of the abomasal nematode Ostertagia circumcincta. AB - Ovine isotype-specific antibody responses to the parasitic larval stages of the abomasal nematode Ostertagia circumcincta were measured in a simple, indirect enzyme-linked immunosorbent assay. Analysis of variance of replicate tests showed that the assay was very reliable. There was substantial variation among individual sheep in their IgA and IgG1 responses even though the sheep had been matched for breed, age and sex, were born on the same farm, were reared identically and had the same history of exposure and challenge with O. circumcincta. The local IgA responses to a somatic extract of fourth-stage larvae were very similar to responses to excretory-secretory products of fourth-stage larvae. The responses to third stage larvae were correlated with the responses to fourth stage larvae. There was a negative correlation between parasite-specific plasma IgG1 and parasite-specific plasma IgA responses. There was only a moderate association between IgA responses in the mucus and the plasma. Therefore, antibody responses measured in plasma cannot be easily extrapolated to antibody responses in the abomasal mucus. PMID- 7483235 TI - Haemonchus placei in calves: effects of dietary protein and multiple experimental infection on worm establishment and pathogenesis. AB - An experiment was conducted to examine the influence of dietary protein and immunisation on parasite establishment and pathogenesis of Haemonchus placei in calves. Four groups of 4-6-month-old worm-free calves (n = 4) were given a low protein diet (LP) containing 213 g crude protein (CP) per head per day or a high protein diet (HP) containing 469 g per head per day CP. Five weeks later, calves in one of the two groups of each dietary treatment were given 50,000 H. placei infective larvae (L3). Twenty-five days later, infection in these groups was terminated by dosing with oxfendazole. This immunisation process was repeated 4 days later. Four days after termination of the second immunisation all calves were challenged with 100,000 L3. Five weeks later, all calves were slaughtered for abomasal worm counts. Worm establishment was lower in the immunised groups; however, only the HP-I group showed a significant reduction (P < 0.05). All calves gained weight during the first 13 week period, and after challenge the non immunised groups lost weight, independent of the level of protein in the diet (P < 0.05). Packed cell volume values for all treatments only dropped after challenge (P < 0.05) and the HP-immunised group presented values significantly higher when compared with the other treatments. All calves were hypoproteinaemic and hypoalbuminaemic at the end of the experiment, regardless of the treatment. Immunised calves showed a normocytic normochromic anaemia, while the non immunised groups presented a microcytic normochromic anaemia. PMID- 7483236 TI - Efficacy of a spring strategic fenbendazole treatment program to reduce numbers of Ostertagia ostertagi inhibited larvae in beef stocker cattle. AB - Efficacy of a spring strategic treatment program with fenbendazole (FBZ) to reduce the accumulation of Ostertagia ostertagi inhibited early fourth-stage larvae (EL4) was investigated in two groups of crossbred beef heifers which were 7-9 months of age and ranged in weight from 155 to 223 kg. The cattle were allocated to groups and treated on 27 April (Day 0). Group 1 calves served as nontreated controls. Group 2 calves were treated with FBZ 10% drench suspension at 5 mg kg-1 on Day 0 and with 6-day courses of FBZ free-choice mineral (to provide 5 mg kg-1 per animal) on Days 28 and 56. Each group grazed on a separate 4.9 ha pasture for 105 days to 10 August. On 10 August the cattle were taken off pasture and each original group was re-allocated to subgroups of three cattle which were treated orally with FBZ (5 mg kg-1), oxfenbendazole (OXF, 4.5 mg kg-1) or left nontreated (CONT). The cattle were necropsied on Days 120 and 121. Mean actual and cumulative fecal egg counts indicated near total suppression of egg output in the strategically treated group (high of 5.2 eggs g-1 feces (EPG) on Day 28). Egg counts of the nontreated group remained above 100 or 200 EPG to Day 63 and then decreased to less than 100. Ostertagia was the predominant genus, followed by Cooperia on most sampling dates. The largest O. ostertagi worm burdens were recovered from the CONT-CONT subgroup; numbers of EL4 ranged from 18,922 to 51,137. Reduction in numbers of EL4 in original controls, treated with FBZ or OXF in August, were low, being 60.2% and 74.3%, respectively. The numbers of O. ostertagi recovered from subgroups originally treated strategically with FBZ were generally lower than in original controls. The largest reduction in O. ostertagi numbers was in the FBZ-CONT subgroup, which was not treated in August. Percent reduction values for O. ostertagi adults, developing L4 (DL4) and EL4 were 84.6%, 96.7%, and 99.0%, respectively. Percent reduction values for adults, DL4 and EL4 in the FBZ-FBZ and FBZ-OXF subgroups were 90.7%, 61.3%, 85.6% and 92.4%, 45.5%, and 73.0%, respectively. Variability in numbers of EL4 acquired during grazing by original controls and strategically treated cattle and variability in efficacy of August treatments was evident as observed in high outlier numbers of worms in all subgroups except the FBZ-CONT subgroup. Liveweights and gains were nearly identical in the two groups on 10 August. PMID- 7483237 TI - Structure and activity of avermectins and milbemycins in animal health. AB - The avermectins and, to a lesser extent, the milbemycins, have revolutionized antiparasitic and antipest control over the last decade. Both avermectins and milbemycins have macrocyclic lactone structures that are superimposable, they are produced by the same genus of soil dwelling organisms, they have the same mode of action, they exert this action against the same nematode/acarine/insect spectrum of targets, and they show the same mechanism-based toxicity in mammals. Reports suggesting that milbemycins have a different mode of action from avermectins with implications that there will be no mutual resistance to the groups have been shown to be false. Contributing to the belief that there were differences in mode of action between the two groups are the vague definitions of resistance presently in use which rely on the ability of the parasite to survive treatment at the manufacturer's recommended use level. More appropriately, drug resistance should be defined as 'a change in gene frequency of a population, produced by drug selection, which renders the minimal, effective dosage previously used to kill a defined portion (e.g. 95%) of the population no longer equally effective'. This type of definition would allow us to detect changes in susceptibility of a population earlier and is essential when comparing different chemicals to determine if there is mutual resistance to them. It is concluded that much effort has been expended by pharmaceutical, government, and academic scientists searching for broad-spectrum second generation avermectin and milbemycin products, but none has exceeded the original avermectin in any fundamental way. The newer avermectin and milbemycin compounds that have appeared claim niches in the market place based on emphasis of certain narrow parts of the overall spectrum. Consequently, there are no second generation avermectins and milbemycins at present and all newer compounds from this mode of action class are viewed as siblings of the first generation. PMID- 7483238 TI - Purification and preliminary characterization of a protease from the excretion secretion products of Trichinella spiralis muscle-stage larvae. AB - A protease from excretion-secretion products of Trichinella spiralis muscle-stage larvae was purified by continuous elution electrophoresis. The state of purification was analyzed electrophoretically using one- and two-dimensional sodium dodecyl sulfate polyacrylamide gel electrophoresis. The purified enzyme was shown to be a single polypeptide with an estimated molecular mass of 35 kDa and isoelectric point of 6.2. Following purification, the enzyme activity was measured by hydrolysis of gelatin, azocoll, azoalbumin, azocasein and collagen as substrates. Maximal azocollytic activity was at pH 5 and a temperature of 37 degrees C. Finally, the proteolytic activity was partially inhibited by N-alpha-p tosyl-lysine chloromethyl ketone, chymostatin and E-64. PMID- 7483239 TI - A method for producing microbe-free Amyloodinium ocellatum (Brown) with Percoll. AB - A method for collecting microbe-free Amyloodinium ocellatum from infected red drum (Sciaenops ocellatus) is described. Surface contamination of trophonts removed from fish by osmotic shock was reduced by washing the preparation with antibiotic containing saltwater and Percoll density gradient purification. The resulting tomont preparation was free of external contamination. Sterile, infective dinospores excysted from the tomonts after 72 h incubation at 26 degrees C in antibiotic containing 30 ppt saltwater. PMID- 7483240 TI - Time series analysis of the prevalence of endoparasitic infections in cats and dogs presented to a veterinary teaching hospital. AB - The apparent prevalence of endoparasitic infections of cats and dogs presented to the small animal Veterinary Hospital of the University of Pennsylvania was measured between 1984 and 1991. Two thousand feline and 8077 canine fecal samples were examined along with 6830 canine blood samples. The overall mean monthly prevalence of feline infections was 16% for ascarids, 0.9% for hookworms, 4.0% for tapeworms, 2.4% for Giardia spp. and 4.2% for coccidia. The overall mean monthly prevalence of canine infections was 5.7% for ascarids, 9.7% for hookworms, 9.7% for whipworms. 1.8% for tapeworms, 4.7% for Giardia spp. and 3.1% for coccidia. There was a significant downward trend in the prevalence of hookworms and heartworms in dogs (P < 0.001 in both cases). There was a significant upward trend in the prevalence of tapeworms in cats (P < 0.05). There were no significant long-term trends in any of the other time series. The smoothed data were analyzed for seasonal trends. None of the autocorrelation analyses gave incontrovertible evidence of seasonality. The repeated peaks at the 6, 12 and 24 month lags in the case of ascarid infections were suggestive of a 12 month seasonality with a peak prevalence in December, but the results were not statistically significant at the 5% level. Hookworms and whipworms in dogs occurred together more than would be expected by chance in 4 out of the 6 years for which data were available. PMID- 7483241 TI - Enzyme-linked immunosorbent assay (ELISA) for the detection of antibodies to Anoplocephala perfoliata in horse sera. AB - A scolex antigen of the horse tapeworm Anoplocephala perfoliata containing at least 14 different proteins was employed in an enzyme-linked immunosorbent assay (ELISA) for detection of antibodies to A. perfoliata in equine sera. The assay was applied to sera from 426 slaughtered horses with different numbers of worms and with varying degrees of intestinal lesions. As measured by the ELISA, there was a very strong effect on the antibody levels both from the number of tapeworms present and from the intestinal lesion score. However, considerable individual variation was observed between horses with similar worm counts. The ELISA values of horses that were either negative for strongyles and tapeworms or positive only for strongyles were similar, whereas both of these groups differed significantly from animals that harboured only A. perfoliata. Thus it seemed that cross reactivity with concurrent nematode infections did not occur. Furthermore, a significant seasonal pattern in antibody levels was observed which reflected the establishment of newly acquired infections. No observations were made of confounding effects owing to age and previous anthelmintic treatment of the horses. It is concluded that the ELISA presented here has a potential for monitoring A. perfoliata infection on a herd level and can be used as a complementary diagnostic tool in epidemiological studies. The present study indicates that transmission of A. perfoliata in Central Sweden occurred during a short period in spring and more consistently in late autumn. PMID- 7483242 TI - Designations F18ab and F18ac for the related fimbrial types F107, 2134P and 8813 of Escherichia coli isolated from porcine postweaning diarrhoea and from oedema disease. AB - The relatedness of the fimbriae produced by eight E. coli strains including type strains with F107 fimbriae, 2134P pili and colonization factor 8813 (preliminary F18), was examined. These strains had been isolated principally from pigs which were affected with postweaning diarrhoea or with oedema disease. The fimbriae were analyzed by means of electron microscopy, slide agglutination, immunofluorescence, immunogold labelling, immuno-diffusion, immunoelectrophoresis and western blot, molecular genetic techniques, and in vitro adhesion. The fimbriae of all the strains were long flexible filaments with a diameter not larger than 4.6 nm showing a zig-zag pattern. Results obtained by the serological techniques confirmed that the fimbriae possessed a common antigenic determinant designated 'a' in addition to a variant-specific determinant designated 'b' or 'c'. Immunoelectron microscopy demonstrated that the determinants 'a' and 'b' or 'a' and 'c' were localized along the same fimbrium. In immunoelectrophoresis, fimbrial extracts of selected strains yielded a single precipitation line towards the cathode. One single major subunit of approximately 15 kDa was recognised in western blots by antisera against the common antigenic determinant and the variant specific determinants. All strains possessed sequences related to gene fedA, coding for the major subunit of fimbriae F107. Two types of fedA-related subunit genes were differentiated, corresponding to the 'ab' and 'ac' types of fimbriae as defined by serological methods. The results demonstrated that F107 fimbriae, 2134P pili and colonization factor 8813 are related, and that two serological variants can be distinguished. We propose designations F18ab (for F107), and F18ac (for 2134P and 8813) in analogy to the nomenclature of F4 fimbriae. PMID- 7483243 TI - Expression of P and type 1 (F1) fimbriae in pathogenic Escherichia coli from poultry. AB - To investigate the expression of P and type 1 (F1) fimbriae in pathogenic avian Escherichia coli, fourteen pap+/fim+ E. coli isolates pathogenic for poultry were grown on four complex or minimal media, and examined for the presence of mannose resistant (MR) and mannose sensitive (MS) hemagglutination (HA), and for P or for type 1 (F1) fimbriae using immunofluorescence, immunodot, and immunoblot. In addition, isolates grown under different culture conditions were examined for adherence to frozen sections of chicken trachea. Twelve of the 14 isolates were divided into three groups based on adhesin expression in the different media. Isolates of all three groups exhibited strong MSHA reactions when cultures were grown serially in static broth, and expressed a subunit protein with an apparent molecular mass of 17 to 18.5 kDa, serologically related to the FIA major fimbrial subunit. There was a good correlation between MSHA and adherence to chicken tracheal sections. Isolates of group I only demonstrated MSHA and expression of F1A fimbriae after growth in static broth. Isolates of group II demonstrated MSHA and expression of F1A fimbriae after growth in all tested media whereas isolates of group III demonstrated expression of F1A fimbriae only after growth in static broth and minimal agar. Only the five group I isolates expressed MRHA associated with P fimbrial adhesins and expressed fimbriae with a major subunit protein of 18 kDa serologically related to the F11 major fimbrial subunit. None of these five isolates grown on complex solid media, where P but not type 1 fimbriae were expressed, adhered to tracheal sections. Results suggest that i) P fimbriae are not readily expressed in vitro by most pap+/fim+ avian E. coli isolates; ii) environmental control of phase variation of type 1 fimbriae differs among pathogenic avian E. coli; and iii) receptors for type 1, but not for P fimbriae, are present in chicken tracheal mucosa. PMID- 7483244 TI - Characterization by restriction endonuclease analysis and DNA hybridization using IS900 of bovine, ovine, caprine and human dependent strains of Mycobacterium paratuberculosis isolated in various localities. AB - DNA of 90 mycobactin-dependent strains of Mycobacterium paratuberculosis, isolated in 9 countries, was digested with restriction endonuclease PstI and hybridized with a DNA fragment containing insertion sequence IS900. Bovine strains (n = 73) were isolated from 61 animals in 17 herds, ovine strains (n = 15) from 13 animals in 3 herds and the set was completed by 1 caprine and 1 human (Linda) strain. Three types, tentatively designated A (n = 37), B (n = 51) and C (n = 2) were differentiated by restriction fragment length polymorphism (RFLP). Of the bovine strains 27, 45 and 1 were classified as belonging to types A, B and C, respectively; of the 15 ovine strains 10 and 5 belonged to types A and B, respectively; the caprine strain belonged to type C. The human strain Linda, isolated in the U.S.A., from a man with Crohn's disease, belonged to B type. A certain degree of type uniformity was observed among strains isolated within one herd in the course of several years, and the prevalence of a single type was also recorded within individual regions of the Czech Republic and Slovakia. Type A was identified in the course of 2 years in a sheep farm with frequent sales and purchases of animals, and type C was demonstrated in a goat kept in the same farm. Differences between RFLP types of the strains isolated from mesenteric lymph nodes and intestinal mucosa were found in one cow and one sheep. Selected strains of M. paratuberculosis RFLP type A (14 strains) and B (18 strains) were digested with restriction endonuclease BstEII. All the strains of A type were classified into C1 group and all the strains of B type into Cx group. PMID- 7483245 TI - Partial characterization of the leukotoxin of Pasteurella haemolytica-like bacteria isolated from swine enteritis. AB - Pasteurella haemolytica-like (PHL) strains isolated from diarrheic pigs are known to produce a leukotoxin that is lethal to ruminant leukocytes. In the present study, 12 PHL strains were screened for leukotoxin production using a tetrazolium dye-reduction assay. Sterile culture supernatant from strain 6213A, the maximum leukotoxin producer, was used as the crude leukotoxin for characterization studies. The leukotoxin was inactivated by heat at 60 degrees C and by trypsin, protease, and amylase. Toxicity was retained over a pH range of 3.0-11.0. The leukotoxin was lethal to polymorphoneutrophils (PMNs) of cattle, sheep, goat, and swine. Chromosomal DNA of all 12 PHL strains hybridized with a 3.9 kb Pasteurella haemolytica A1 leukotoxin probe, indicating similarities between the leukotoxin genes of P. haemolytica and PHL strains. PMID- 7483246 TI - Tetracycline resistance determinants among streptococci of serological group G and L. AB - In the present study 56 streptococci of serological group G and L isolated from various animal species and from humans were investigated for tetracycline and minocycline resistance and for the presence of genes conferring this combined resistance. Among the 45 group G streptococci, 2 isolates from dogs, 3 from cattle and 2 from humans, respectively, as well as all 11 group L streptococci isolated from cattle, pigs or poultry were resistant to tetracycline and simultaneously to minocycline. The restriction endonuclease digested and blotted DNA-preparation of the tetracycline-and minocycline resistant group G streptococci from dogs and humans hybridized with the tet (M) gene probe, those from bovines with the tet (O) gene probe. Six group L streptococci carried the gene tet (M), whereas 5 isolates harboured the gene tet (O). The tet (M)-and tet (O) gene probes recognized complementary sequences on EcoRI-fragments of various sizes. PMID- 7483247 TI - Infection of cultured rat enterocytes by Ileal symbiont intracellularis depends on host cell function and actin polymerisation. AB - The mechanisms of entry of Ileal symbiont intracellularis into IEC-18 rat enterocyte cells and subsequent bacterial proliferation were examined in centrifuge-assisted and static infections. Live, oxygen or neomycin damaged, and formalin killed bacteria, each rapidly entered viable cells. Live or damaged bacteria did not enter cells nor proliferate within cells after static infection of cells cooled to 5 degrees C. Infection of cells was greatly reduced at 20 degrees or 32 degrees compared to infection at 37 degrees C. Centrifuge-assisted infection was also reduced by chilling the cells. Cytochalasin D but not B inhibited the entry process indicating an actin-dependent infection, although other pathways may also be involved in centrifuge-assisted infections. Drugs capable of modifying cell membrane charge, heparin receptors or trypsin-labile proteins were all inactive in preventing or enhancing infection. We therefore conclude that infection of enterocytes by IS intracellularis is dependent on host cell activity and actin polymerization, but is independent of bacterial viability. PMID- 7483248 TI - Prevalence of neutralizing antibodies to 9 rotavirus strains representing 7 G serotypes in sheep sera. AB - Neutralizing antibodies to 9 rotavirus strains representing serotypes G1, G3, G5, G6, G8, G9, and G10 were investigated in 212 ovine serum samples from 3 age groups, 1-week-old lambs, 2- to 3-months-old lambs and adult sheep. All sera from 1-week-old lambs had neutralizing antibodies to all 9 rotavirus strains. Both neutralizing antibody titers and prevalences to all 9 strains markedly decreased in the 2- to 3-months-old lamb group and increased again in the adult sheep group. Also, adult sheep sera neutralized a larger number of rotavirus strains than 2- to 3-months-old lamb sera. The highest neutralizing antibody titers and prevalences were found to strains B223 and K923, representing serotype G10, to strain RRV, representing serotype G3, and to strain NCDV, representing serotype G6, indicating that these could be the predominant 3 rotavirus serotypes in Spanish sheep. The rotavirus serotypes infecting sheep observed by us differ from those described for cattle, where G6 is the most prevalent serotype followed by G10, and G3 has been seldom found. Very low prevalences were observed for strains WA and OSU representing serotypes G1 and G5 respectively, suggesting that they probably do not infect sheep and neutralizing antibodies found are derived from heterotypic responses to other serotypes. Intermediate prevalences and titers were found to strains UK (serotype G6), 69M (serotype G8) and WI61 (serotype G9). Neutralizing antibodies distinguished between different strains sharing their VP7 specificity: B223 and K923, a bovine and an ovine serotype G10 strains, and NCDV and UK, two serotype G6 bovine rotavirus strains with different VP4 antigen. PMID- 7483249 TI - Genetic and antigenic characterization of caev (caprine arthritis-encephalitis virus) recombinant transmembrane protein. AB - The env gene fragment of an Italian strain of Caprine Arthritis Encephalitis virus (CAEV) coding for the hydrophilic region of transmembrane protein was amplified, cloned and expressed in prokaryotic system as fusion protein with glutathione-S-transferase. Sequence analysis revealed 63 to 66% amino acid homology, when compared with three ovine lentiviruses and 83% when compared with one caprine lentivirus. The recombinant transmembrane protein was efficiently expressed, purified under denaturing conditions and used as antigen in western blotting and ELISA. Sera from clinically diseased goats strongly reacted in western blotting and naturally infected animals seroconverted between 20 and 33 weeks of age. An indirect ELISA performed with this antigen showed improved sensitivity in comparison with agar gel immunodiffusion test. Our results confirm that transmembrane protein is an important immunological marker in CAEV infection and its use as antigen may enhance the validity of serological diagnosis of Caprine Arthritis Encephalitis. PMID- 7483250 TI - Characterization of camelpoxvirus isolates from Africa and Asia. AB - Five orthopoxvirus isolates of camels from different geographic regions of Africa and Asia were analysed with respect to their biological and genomic attributes. The behaviour of the isolates in various cell cultures, the type of pock lesions on the chorioallantoic membrane of embryonated chicken eggs, and the respective ceiling temperatures were determined. Additionally, physical maps for restriction endonucleases HindIII and XhoI were established. The data obtained from biological assays and DNA analyses demonstrated minor differences between the five isolates. However, these findings confirm previous reports suggesting that orthopoxviruses of camels constitute a separate species within the genus Orthopoxvirus. PMID- 7483251 TI - Cross-protection experiments in pigs vaccinated with Actinobacillus pleuropneumoniae subtypes 1A and 1B. AB - Cross-protection experiments were conducted to determine whether antigenic differences located within the lipopolysaccharides (LPS) of Actinobacillus pleuropneumoniae subtypes 1A and 1B were important with respect to the efficacy of whole cell, formalin-inactivated bacterins. Based on clinical signs, lung lesions scores and mortality rates, pigs immunized with A. pleuropneumoniae subtype 1A were partially protected against severe challenge with both subtypes 1A and 1B. In contrast, 1B vaccinated pigs were not protected against severe challenge with subtype 1A but were partially protected against 1B challenge. Cross-reactive serum antibody levels were measured with an ELISA using outer membranes of subtype 1A or 1B as the coating antigen. Serum antibodies were detected against both subtypes within 2 weeks after the first immunization. Antibody levels increased with time and were generally higher against the homologous subtype coating antigen. We conclude that antigenic variation within a capsular serotype, due to antigenic variation within LPS, can result in the failure of whole cell bacterins to provide protection against challenge with the same capsular serotype. This lack of cross-protection within a capsular serotype provides partial explanation for vaccination failures observed under field conditions. PMID- 7483252 TI - Immunization trial of cats with a replication-defective adenovirus type 5 expressing the ENV gene of feline immunodeficiency virus. AB - Our aim was to develop a recombinant replication-defective adenovirus suitable for the vaccination of cats against feline immunodeficiency virus. We first demonstrated that this vector was able to transfer a marker gene (E. coli beta galactosidase) in feline cells in vitro. We then constructed an adenovirus type 5 expressing the Feline Immunodeficiency Virus (FIV) envelope (ENV) gene of the Wo isolate in the absence of the rev gene (Ad-ENV-Wo). Ad-ENV-Wo was then tested in four cats in a 3 injections scheme (at day 0, day 30 and day 210). Four other control cats received Ad-gp50, a similar recombinant adenovirus expressing gp50 (Ad-gp50) of pseudorabies virus (PRV). Viruses were formulated in two different kind of oil adjuvants (water/oil and water/oil/water), a protocol previously shown to enhance the immune response against the virus-induced protein. The control cats developed neutralizing antibodies against PRV, demonstrating the potency of recombinant human adenovirus 5 (Ad5) as a vector in cats. Antibody responses appeared after the first injection and were higher with the water/oil/water formulation than with the water/oil controls. However, none of the four cats vaccinated with Ad-ENV-Wo developed antibodies against two peptides of the envelope protein. Animals were challenged with 20 infectious doses 50% of the strain Wo. All of them developed antibodies against FIV within 4 to 5 weeks, and FIV virus could be isolated from all. PMID- 7483253 TI - Comparison of packaging strategy in retroviruses and pararetroviruses. AB - Reverse transcription is not solely a retroviral mechanism. Animal hepadnaviruses, plant caulimoviruses, and badnaviruses have a RNA intermediate which is reverse transcribed into double-stranded DNA genome. Based on this fact, these three groups of viruses have been regrouped under the name of pararetroviruses. Although each one has developed its own strategy to assure an efficient packaging of their genome, it is clear that they have adopted a strategy where encapsidation prepares for initiation of reverse transcription. This is discussed in this review. PMID- 7483255 TI - Coexistence of several novel hantaviruses in rodents indigenous to North America. AB - Three genetically distinct members of the Hantavirus genus have been detected in Nevada rodents by RT-PCR and nucleotide sequence analysis. These include Sin Nombre (SN), El Moro Canyon (ELMC), and Prospect Hill (PH)-like viruses which are primarily associated with Peromyscus maniculatus (deer mouse), Reithrodontomys megalotis (western harvest mouse), and Microtus spp. (voles), respectively. Although this region of the United States is ecologically diverse, rodents infected with different hantaviruses appear to coexist in several different geographical and ecological zones. In two widely separated states, Nevada and North Dakota, PH-like viruses are present in three different species of vole. In addition, ELMC-like virus has been detected in both R. megalotis and M. montanus (mountain vole). SN virus is a cause of hantavirus pulmonary syndrome throughout much of the United States. SN virus RNA is found in 12.5% of P. maniculatus in Nevada and eastern California. Two lineages of SN virus coexist in this region and differ from SN viruses originally found in infected rodents in New Mexico, Arizona, and Colorado. These data show the complexity of hantavirus maintenance in rodents. Distinct hantaviruses or virus lineages can coexist either in different or the same rodent species and in either different or the same geographic or ecological zones. PMID- 7483256 TI - Pathogenesis and immunogenicity of bovine adenovirus type 3 in cotton rats (Sigmodon hispidus). AB - Intranasal inoculation of cotton rats (Sigmodon hispidus) with 10(8) PFU of bovine adenovirus type 3 (BAd3) resulted in limited virus replication in the lung and trachea. Histopathological changes in the lungs were characterized by necrosis and hyperplasia of bronchiolar epithelium, eosinophilic intranuclear inclusions, pneumocyte type II hyperplasia in the alveoli, and mild peribronchiolar and perivascular lymphocytic infiltration. Immunohistochemically, viral antigens were observed more frequently in bronchiolar epithelial cells than in alveolar cells in cotton rat lung sections stained using a rabbit anti-BAd3 serum. Bronchiolar epithelial changes, intranuclear inclusion bodies, type II pneumocyte proliferation, peribronchiolar infiltration, and immunohistological staining were maximum at Day 3 or Day 4 postinoculation, whereas perivascular infiltration was first observed at Day 8 postinoculation. In addition to the histological study of the pathogenesis of BAd3 infection, we monitored the BAd3 specific immune response in cotton rats. Anti-BAd3 IgG and virus neutralizing antibodies were detected in sera, whereas anti-BAd3 IgA antibodies were found in the sera, lung, and nasal washes. Our results suggest that the cotton rat can serve as a useful small-animal model for investigating the pathogenesis of BAd3 infection, as well as immune responses to BAd3 recombinant virus vaccines. PMID- 7483254 TI - Bacteriophage P2: genes involved in baseplate assembly. AB - The sequences of two previously defined tail genes, V and J, of the temperate bacteriophage P2, and those of two new essential tail genes, W and I, were determined. Their order is the late gene promoter, VWJI, followed by the tail fiber genes H and G, and a transcription terminator. The V gene product is the small spike at the tip of the tail, and the J gene product lies at the edge of the baseplate. The W gene product may be homologous to the product of gene 25 of T4 phage, which is part of the T4 baseplate. A temperature-sensitive mutation in gene V affects satellite phage P4 production more than it affects the production of P2 helper phage. P4 mutations that partially compensate for this defect of gene V lie in the P4 capsid size determination gene, sid. PMID- 7483257 TI - Identification of critical amino acids within the foot-and-mouth disease virus leader protein, a cysteine protease. AB - The Leader protein of foot-and-mouth disease virus (FMDV) is the first component of the virus polyprotein. It is synthesized in two forms, Lab and Lb, both of which display the ability to cleave the L/P1 junction in trans and to induce the cleavage of the cap-binding complex component eIF-4G (p220). The L protease has weak homology to the family of cysteine proteases, which have a catalytic dyad composed of a cysteine and a histidine. Mutations have been introduced into FMDV cDNA to modify each of the four cysteine residues and the three conserved histidine residues within the Lb species. The activities of the mutant L proteins have been determined. Modification of a single cysteine residue (residue 51) or of a single histidine residue (residue 148) abolished the abilities of L to cleave the L/P1 junction and to inhibit cap-dependent protein synthesis. In contrast, modification of each of the other cysteine residues and other conserved histidine residues had no apparent effect on these activities. PMID- 7483258 TI - Epstein-Barr virus infection of CR2-transfected epithelial cells reveals the presence of MHC class II on the virion. AB - Epithelial cell lines transfected with the Epstein-Barr virus (EBV) receptor CR2 are susceptible to infection by EBV. Following infection with certain EBV strains we found that these cells became positive for MHC class II. The class II was confirmed as being of viral and not target cell origin by immunostaining with HLA specific monoclonal antibodies. Electron microscopic immunogold staining confirmed the presence of MHC class II on the surface of the virion. While some MHC class I was also found on the EB virion, other cell surface molecules were absent. Dual color immunofluorescence and confocal microscopy analysis demonstrated colocalization of class II with EBV-encoded structural proteins (MA and VCA) in infected epithelial cells. However, preincubation of EBV with antibodies against either MHC class I or MHC class II failed to affect either EBV binding or EBV infection. The presence of MHC on the surface of the EB virion may be a consequence of the intracellular pathways through which productive virus exits from the cell and may influence the target cell tropism of EBV. PMID- 7483259 TI - Both virus and host components are important for the manifestation of a Nef- phenotype in HIV-1 and HIV-2. AB - While it has been demonstrated that the Nef protein of simian immunodeficiency virus is obligatory for the establishment of high viral loads and the development of simian AIDS in rhesus macaques, demonstrating a critical role for the human immunodeficiency virus (HIV) Nef protein in tissue culture has been elusive. Data have been contradictory as to whether Nef has a negative or positive influence on in vitro virus replication. In an attempt to define a role for Nef during virus propagation in tissue culture and to obtain virus-host systems that could distinguish between the Nef mutant and wild-type viruses, we have introduced mutations into the nef genes of infectious molecular clones of three HIV-1 strains and two isolates of the HIV-2ROD strain and have investigated the capacity of viruses derived from them to infect a number of CD4-positive T-cell lines and peripheral blood mononuclear cells (PBMC). Mutating the nef gene of all viruses had a modest negative effect on virus production in activated PBMC. In some T-cell lines with some viruses, the effects were severe, and little or no Nef mutant virus could be detected. In other cell lines, the result of mutating the nef gene either had no effect or had a slight negative effect on the replication kinetics. Therefore, whether the consequences of loss of Nef activity can be demonstrated in vitro depends on both the particular virus and the host cell used, suggesting that Nef is exerting its activity on some cellular pathway. In addition, we describe the construction and properties of hitherto unreported infectious molecular clones of the ROD strain of HIV-2. PMID- 7483260 TI - The flavivirus 3'-noncoding region: extensive size heterogeneity independent of evolutionary relationships among strains of tick-borne encephalitis virus. AB - The sequences of the 3'-noncoding regions (NCR) of 12 strains of tick-borne encephalitis (TBE) virus were analyzed and found to vary in length from 350 to approximately 750 nucleotides. The size heterogeneity is restricted to a variable region following the stop codon, whereas the most 3'-terminal 350 nucleotides form a highly conserved core element containing several potentially important sequence motifs and secondary structure elements. A homoadenosine tract previously thought to form the 3'-terminus of some TBE virus strains was now shown to be an internal part of the variable region of certain strains. The strains included in this study were isolated from both humans and ticks over a time period of more than 40 years at various locations throughout the entire endemic area of TBE virus, but there was no correlation between these parameters and the observed lengths of the 3'-NCRs. Identity data calculated from common 3' NCR sequences and also from short sections of the open reading frame indicated that coding and noncoding sequences were linked during evolution, but the lengths of the 3'-NCRs were independent of these relationships. These observations together with detailed analyses and alignments of the sequences suggest that the variable region was originally acquired through duplication and recombination events, but--much more recently during evolution--various portions of this region were lost again, resulting in the now observed heterogeneous 3'-NCRs. PMID- 7483261 TI - Identification of V3 mutations that can compensate for inactivating mutations in C4 of simian immunodeficiency virus. AB - A valine to isoleucine substitution at position 322 within variable region 3 (V3) of envelope of simian immunodeficiency virus was previously shown to compensate for an inactivating valine to glycine mutation at position 448 in constant region 4 (C4) (Morrison et al., Virology 195, 167-174, 1993). Cloned DNA fragments with inactivating C4 mutations were combined with complex mixtures of mutant V3 sequences, and full length genomes were transfected into COS-1 cells. By cocultivating transfected cells with CEM x 174 cells, we were able to identify two additional compensatory V3-C4 combinations. Changing 334 proline to leucine compensated for an inactivating 428 asparagine to lysine mutation and changing 324 isoleucine to leucine compensated for an inactivating 448 valine to glycine mutation. The double mutants replicated efficiently in CEM x 174 cells, rhesus monkey peripheral blood mononuclear cells, and the continuously growing rhesus monkey T cell line 221. Surprisingly, the 324 I-->L and 33 P-->L mutations by themselves impaired SIVmac239 wild-type replication in CEM x 174 cells. These results confirm the cooperation between V3 and C4 sequences and they define additional specific residues participating in this cooperation. PMID- 7483262 TI - Biochemical analysis of the major vaccinia virus envelope antigen. AB - The major envelope antigen of vaccinia virus is an acylated protein of M(r) 37,000 (p37K) which is required for the formation of extracellular enveloped virions (EEV). Despite its important role in the wrapping process, p37K has not been studied in much detail. In order to better characterize this protein we have undertaken a detailed biochemical analysis. Sodium carbonate treatment showed that p37K is tightly bound to the viral envelope. Its resistance to proteinase K digestion indicates that it is not exposed on the surface of EEV but lines the inner side of the envelope. Since p37K does not contain a signal peptide characteristic of most membrane proteins, we examined the possibility that the protein acquires its membrane affinity through the addition of fatty acids. Indeed, Triton X-114 phase partitioning experiments demonstrated that p37K is hydrophobic when acylated, but hydrophilic in the absence of fatty acids. Three other viral proteins have been shown to be required for virus envelopment and release from the host cell and we therefore tested whether p37K interacts with viral proteins. In EEV and in absence of reducing agents, an 80-kDa complex reacting with an anti-37K antiserum was found. Analysis of this complex showed that it most likely consists of a p37K homodimer. Interestingly, only a small amount of p37K occurs as a complex, most of it is present in the viral envelope as monomers. PMID- 7483263 TI - Identification of regions on the hemagglutinin-neuraminidase protein of human parainfluenza virus type 2 important for promoting cell fusion. AB - The hemagglutinin-neuraminidase (HN) and fusion (F) glycoproteins of two paramyxoviruses, human parainfluenza virus type 2 (PIV2) and simian virus 41 (SV41), were expressed in HeLa cells by transfecting with recombinant plasmid harboring each glycoprotein gene. Expressed F proteins could not induce cell fusion by themselves, but evoked prominent cell fusion when coexpressed with homologous HN proteins. It was also proved that PIV2 HN protein could weakly promote SV41 F-mediated cell fusion. By analyzing the fusion-promoting function of chimeric HN proteins of PIV2 and SV41, it was revealed that the N-terminal region (about 16% of total amino acids) of either PIV2 HN or SV41 HN protein could define the type-specific fusion-promoting function for homologous F protein. Analyses of additional chimeras indicated that the N-terminal region in PIV2 HN protein (designated region I, consisting of 94 amino acids) could be reduced to a 58-amino-acid region (region I') which was located at the membrane proximal end of the ectodomain. Furthermore, PIV2 HN protein proved to promote cell fusion mediated by PIV4A F protein. Unexpectedly, analyses of another set of chimeras revealed that the promoting function of PIV2 HN protein for PIV4A F mediated cell fusion was not merely carried by its region I but also by another region ranging from residue 148 to 209 (region II). Finally, it was indicated that regions I' (in the presumed stalk domain) and II (in the globular head) in PIV2 HN protein might play important roles in promoting cell fusion mediated by the F proteins. PMID- 7483264 TI - Beet western yellows luteovirus capsid proteins produced by recombinant baculoviruses assemble into virion-like particles in cells and larvae of Bombyx mori. AB - We subcloned various constructs of the beet western yellows luteovirus (BWYV) 3' proximal genes into Bombyx mori nuclear polyhedrosis virus (BmNPV) transfer vectors and constructed recombinant BmNPVs. The recombinant BmNPVs were used to infect BmN cells and B. mori larvae. Protein expression was analyzed by SDS-PAGE and immunoblot analysis, and the BWYV-encoded capsid protein (CP), P19, readthrough protein (RT), and a modified capsid-readthrough protein (CP-RT*) were detected in BmN cells infected by the respective BmNPVs. However, we detected only the BWYV CP and P19 in BmN cells infected with a recombinant BmNPV containing all three BWYV 3' proximal genes. BmN cells and fat body cells from B. mori larvae infected with recombinant BmNPVs were analyzed by transmission electron microscopy and immunogold labeling. Particles which morphologically and serologically resembled BWYV virions were detected in the nuclei of BmN cells and B. mori larval fat body cells infected with recombinant BmNPVs producing either the BWYV CP alone or BWYV CP plus a BWYV CP-RT*. Similar particles were not detected in cells infected with a control BmNPV or recombinant BmNPV producing only CP-RT*. PMID- 7483265 TI - Identification and site-directed mutagenesis of the primary (2A/2B) cleavage site of the hepatitis A virus polyprotein: functional impact on the infectivity of HAV RNA transcripts. AB - The junction between 2A and 2B proteins of the hepatitis A virus (HAV) polyprotein is processed by the virus-encoded 3C protease to liberate the precursor for capsid proteins, but details of this cleavage remain poorly defined. We identified the location of this primary cleavage by a novel approach involving expression of HAV polypeptides in eukaryotic cells via recombinant vaccinia viruses. A substrate polyprotein spanning the putative HAV 2A/2B site was fused at its C-terminus to a poliovirus VP1 reporter sequence. This substrate was cleaved efficiently in trans by protease 3C derived from another recombinant vaccinia virus expressing a 3C precursor protein. N-terminal sequencing of the 2B poliovirus VP1 fusion product identified the site of cleavage as the Gln836/Ala837 dipeptide, 144 residues upstream of the originally predicted site. Two mutations were introduced at the P1 position of the 2A/2B site. Gln836-->Asn, and Gln836-->Arg. Asn substitution at the P1 residue reduced the efficiency of cleavage in the vaccinia expression system and resulted in a small replication focus phenotype of virus rescued from infectious HAV RNA transcripts. Arg substitution abolished cleavage and was lethal to HAV replication. In addition to identifying the site of the primary HAV polyprotein cleavage, these results shed light on the in vivo specificities of the HAV 3C protease. PMID- 7483266 TI - Origin and molecular changes associated with emergence of a highly pathogenic H5N2 influenza virus in Mexico. AB - In October of 1993, there was decreased egg production and increased mortality among Mexican chickens, in association with serologic evidence of an H5N2 influenza virus. First isolated from chickens in May of 1994, after spreading widely in the country, the virus caused only a mild respiratory syndrome in specific pathogen-free chickens. Because eradication of the virus by destruction of infected birds posed major obstacles to the poultry industry in Mexico, we were able to conduct a "field experiment" to determine the fate of an avirulent virus after repeated cycles of replication in millions of chickens. By the end of 1994, the virus had mutated to contain a highly cleavable hemagglutinin (HA), but remained only mildly pathogenic in chickens. Within months, however, it had become lethal in poultry. Nucleotide sequence analysis of the HA cleavage site of the original avirulent strain revealed R-E-T-R, typical of avirulent viruses and unlike the K-K-K-R sequence characterizing viruses responsible for the 1983 outbreak in poultry in the United States. Both mildly and highly pathogenic isolates contained insertions and a substitution of basic residues in the HA connecting peptide, R-K-R-K-T-R, which made the HA highly cleavable in trypsin free chicken embryo fibroblasts. Phylogenetic analysis of the HA of H5 avian influenza viruses, including the Mexican isolates, indicated that the epidemic virus had originated from the introduction of a single virus of the North American lineage into Mexican chickens. This sequence of events demonstrates, apparently for the first time, the stepwise acquisition of virulence by an avian influenza virus in nature. PMID- 7483267 TI - Interplay between a new HNF3 and the HNF1 transcriptional factors in the duck hepatitis B virus enhancer. AB - We identified a new hepatocyte nuclear factor 3 (HNF3) binding site in the DHBV enhancer. This site is close to the hepatocyte nuclear factor 1 (HNF1) binding site, responsible for most of the enhancing activity. No differences in the migrating properties were found between this new site and the two other HNF3 sites recently described in this enhancer. Factor HNF1 strongly inhibits binding of the HNF3 factor in this newly characterized site. The two factors were never detected simultaneously on the DNA fragment, even when their respective concentrations were modified. Competition persisted after enlarging by 5 and 10 nucleotides the space between the two sites. On the contrary, when the HNF3 binding site was changed into the perfect consensus site, binding of the HNF3 factor was not inhibited any longer by HNF1 and a supershift, corresponding to the binding of both factors, was observed. Thus a limited mismatching appears to modulate the interaction between transcriptional proteins and DNA and allows a second transcriptional protein to interplay with the former one. PMID- 7483268 TI - Cooperative neuraminidase activity in a paramyxovirus. AB - Paramyxoviruses possess neuraminidase (NA) activity, i.e., the ability to cleave sialic acid from membrane-bound and soluble glycoconjugates. The activity is associated with a homotetrameric, surface glycoprotein spike, called the hemagglutinin-neuraminidase. This structure also mediates viral attachment to sialic acid-containing receptors and constitutes the major neutralizing antigen for the virus. Cooperativity has been demonstrated for the NA activity of an isolate of one of the paramyxoviruses, Newcastle disease virus. Although all known viral NA proteins are homooligomeric, this is the first demonstration of cooperativity in this family of proteins. A variant virus, selected by escape from neutralization by a monoclonal antibody thought to bind close to the NA active site, has lost cooperativity. Conversion to the noncooperative state correlates with increases in both avidity for cellular receptors and fusogenic activity. PMID- 7483269 TI - In vitro infection of bone marrow-adherent cells by human immunodeficiency virus type 1 (HIV-1) does not alter their ability to support hematopoiesis. AB - As an attempt to elucidate the pathogenesis of human immunodeficiency virus type 1 (HIV-1)-related cytopenia, the effects of infection of long-term primary bone marrow culture (LTBMC)-derived adherent cells on hematopoiesis were investigated. Productive infection could then be established only when using monocytotropic strains HIV-1Ba-L, HIV-1Ada, and HIV-1JR-FL but not with lymphocytotropic strain HIV-1LAI. Culture supernatants were tested for major cytokines involved in the regulation of hematopoiesis: neither IL-3 nor GM-CSF were detectable in the infected or noninfected cultures; in contrast, TGF-beta, TNF-alpha, MIP-1 alpha, Steel Factor, and IL-6 were detected at all times in established LTBMCs, but their levels were not consistently altered by virus replication. In vitro functional analysis by colony and long-term culture assays showed that HIV-1 infection failed to alter either the kinetics or the number of hematopoietic progenitors produced by the stromal layers; it did not interfere with the clonogenicity of exogeneous CD34+ cells in semisolid assays, and no difference was observed relative to the controls when HIV-1-infected stromal layers were tested for their ability to sustain long-term hematopoiesis. These results show that productive and sustained virus replication in the macrophage component of LTBMCs does not significantly alter the profile of major cytokines involved in regulating hematopoiesis, nor is it sufficient by itself for altering in vitro hematopoiesis under the baseline conditions used. PMID- 7483270 TI - Immune protection conferred by the baculovirus-related glycoprotein of Thogoto virus (Orthomyxoviridae). AB - The coding region of segment 4 of Thogoto (THO) virus, a tick-borne member of the Orthomyxoviridae, was expressed in a baculovirus system under the control of the polyhedrin promoter. This construct expressed authentic envelope glycoprotein as determined by size and antigenic reactivity with a panel of monoclonal antibodies (MAbs). Immunization of hamsters with Spodoptera frugiperda (Sf21) cells infected with the recombinant baculovirus induced neutralizing and protective antibodies against virus challenge; control hamsters developed clinical disease with high titer viremia 3 days postchallenge. In contrast to hamsters, guinea pigs are comparatively resistant to THO virus infection but support nonviremic transmission between cofeeding infected and uninfected ticks. However, when uninfected ticks fed on guinea pigs immunized with the baculovirus recombinant, only 2% became infected following virus challenge of the animals compared to 26% of ticks on control nonimmune guinea pigs. Furthermore, neutralizing MAbs specific for THO viral glycoprotein protected mice against lethal challenge with THO virus; nonneutralizing MAbs specific for the glycoprotein, which inhibit THO viral agglutinating activity, did not induce a protective response. Thus at least in the murine model, protective immunity is conferred by antibodies directed against the neutralizing epitope(s) of the baculovirus-related glycoprotein of THO virus. PMID- 7483271 TI - Antibody-enhanced binding of dengue-2 virus to human platelets. AB - The mechanisms underlying severe thrombocytopenia in dengue hemorrhagic fever/dengue shock syndrome (DHF/DSS) are not completely understood. We present here the first evidence that dengue type 2 virus binds to human platelets only in the presence of virus-specific antibody, supporting a role for immune-mediated clearance of platelets in the pathogenesis of thrombocytopenia in DHF/DSS. Antibody-enhanced binding of virus of platelets was also demonstrated with a panel of eight murine monoclonal antibodies specific for the dengue E protein. The degree of binding was dependent on the antibody used but not on the antibody IgG subclass, indicating that factors other than the platelet Fc receptor are involved in binding of virus-antibody complexes to the platelet surface. Confirmation that antibody-dependent virus binding to platelets is not primarily mediated by the platelet Fc receptor was obtained by demonstrating good binding even when platelets were pretreated with the Fc gamma RII-specific antibody IV.3. PMID- 7483272 TI - Equine herpesvirus 1 gene 12, the functional homologue of herpes simplex virus VP16, transactivates via octamer sequences in the equine herpesvirus IE gene promoter. AB - The HSV-1 transactivator of immediate-early (IE) gene expression, VP16, has several functional homologues among the alphaherpesviruses which have not yet been extensively studied in relation to their modes of action. To date, nothing is known of the exact sites or mechanism of interaction of the equine herpesvirus type 1 (EHV-1) homologue, the gene 12 protein, with the EHV-1 IE promoter. We show that the gene 12 protein utilises the promoter proximal region of the IE gene to induce activation and identify four potential octamer DNA binding sites within that region. Although there was divergence from its consensus, Oct-1 bound to each of these sites in an in vitro complex formation assay, and in the presence of the gene 12 product a second complex of slower migration, which was also dependent on Oct-1, was detected. When each site was inserted into a basal promoter, two conferred activation by gene 12 with a resulting increase in expression of up to 50-fold compared to basal levels. These results show that, despite the differences between the two proteins, the mechanism of interaction of the gene 12 protein with its target is analogous to that of VP16. PMID- 7483273 TI - Structural requirements in the membrane-spanning domain of the paramyxovirus HN protein for the formation of a stable tetramer. AB - The paramyxovirus hemagglutinin-neuraminidase (HN) is a type II homotetrameric integral membrane glycoprotein composed of a pair of disulfide-linked dimers that are held together by noncovalent bonds. To determine the role of the internal uncleaved signal-anchor (S/A) domain in stable tetramer formation, cDNA-derived HN mutants containing S/A substitutions were expressed in HeLa cells. The assembly into tetramers and ER-to-Golgi transport of the proteins were examined by sucrose gradient sedimentation and by endoglycosidase treatment. A leucine scanning substitution analysis of the 19-residue S/A identified 2 polar residues (Ser 31 and Tyr 36) in the C-terminal end of the S/A that were important for the formation of a stable tetramer. While Ala, Cys, and Gly could functionally replace Ser 31 in the formation of a stable tetramer, substitution with Leu or Phe resulted in mutants that were detected as disulfide-linked dimers. These results indicate that a small amino acid in position 31, rather than a specific residue per se, is an important assembly requirement in the S/A. In contrast to the size requirement for position 31, the conservative substitution of Tyr 36 with Phe produced an HN mutant that sedimented as a mixture of dimers, tetramers, and higher order oligomers, suggesting that proper assembly requires a Tyr in this position. The S/A mutants that were detected as disulfide-linked dimers showed only a slight reduction in ER-to-Golgi transport (approximately 50% of WT), consistent with the proposal that the S/A substitutions had affected tetramer stability and not the formation of a transport-competent oligomer. These data indicate that there are different structural requirements for two positions in the C-terminal region of the HN S/A for the assembly of a stable tetramer. PMID- 7483274 TI - Mobilization of an ERV9 human endogenous retroviral element during primate evolution. AB - ERV9 is a low repeated family of human endogenous retroviral elements, which has close to 50 members, in addition to at least 4000 solitary LTRs. Previous work has shown that randomly selected LTRs can promote transcription of reporter genes, raising the possibility that these sequences may affect the expression of adjacent cellular genes. We report here the structural organization in different primate species of a zinc-finger coding gene whose expression is driven in humans by a solitary ERV9-LTR promoter. Using a PCR strategy and library screening, we were able to trace the origin of the insertion event in the primate lineage and to evaluate the impact of this event on gene structure. Our findings indicate that the integration of the ERV9 element occurred after the split of orangutang from the great apes, but before the divergence of the gorilla lineage. These results suggest that ERV9 elements have been mobile within the primate lineages and may still be active in humans. PMID- 7483275 TI - A novel carbohydrate addition site on the hemagglutinin protein of a highly pathogenic H7 subtype avian influenza virus. AB - The highly pathogenic (HP) avian influenza isolate, A/Fowl/Victoria/76 (H7N7), contains two naturally occurring hemagglutinin (HA) variants. The two hemagglutinin proteins differ only in the possession of a potential asparagine linked glycosylation site at amino acid position 188-190, which is near the proposed receptor binding region of the HA. Expanded virus plaques which possess the addition site exhibit more slowly migrating HA1 subunits and are significantly more lethal in chickens than those which lack the site. When artificial mixtures of the two variants were inoculated in birds, as few as 1 in 1000 particles containing the glycosylation site was sufficient to exhibit 100% lethality in birds. The data raise the possibility that presence of carbohydrate near the receptor site on the H7 avian influenza virus hemagglutinin may influence virulence. PMID- 7483277 TI - Tobamovirus and dianthovirus movement proteins are functionally homologous. AB - The movement proteins (MPs) of tobacco mosaic tobamovirus (TMV) and red clover necrotic mosaic dianthovirus (RCNMV) enlarge plasmodesmata size exclusion limits, transport RNA from cell to cell, and bind nucleic acids in vitro. Despite these functional similarities, they have no sequence homology. However, they do appear to have similar secondary structures. We have used transgenic plants expressing either the TMV MP or the RCNMV MP, and a chimeric TMV that encodes the RCNMV MP as its only functional MP gene, to demonstrate that the MPs of TMV and RCNMV are functionally homologous. Further, both TMV and RCNMV can act as helper viruses to allow the cell-to-cell movement of the heterologous movement-defective viruses. These data support the conclusion that, despite other differences, such as particle morphology, host range, and sequence, TMV and RCNMV share a common mechanism for cell-to-cell movement. PMID- 7483276 TI - Identification and transcriptional analysis of a 3'-coterminal gene cluster containing UL1, UL2, UL3, and UL3.5 open reading frames of bovine herpesvirus-1. AB - We have identified and sequenced 3113 nucleotides located at the right end of the HindIII L fragment of the bovine herpesvirus-1 genome from map units 0.712 to 0.734. Analysis of the sequence identified four open reading frames (ORFs) which are designated UL1, UL2, UL3, and UL3.5 based on their homology with proteins of herpes simplex virus-1 (HSV-1), pseudorabies virus (PRV), equine herpesvirus-1, and varicella-zoster virus. The UL1 ORF of 158 amino acids exhibits limited homology with UL1 (glycoprotein gL) of HSV-1 (27%) and PRV (21%). The UL2 ORF of 204 amino acids shows significant homology to UL2 (uracil-DNA glycosylase) of HSV 1 (68%) and PRV (75%). The UL3 ORF of 204 amino acids shows significant homology to UL3 (nuclear phosphoprotein) of HSV-1 (62%) and PRV (53%). The UL3.5 ORF of 126 amino acids shows limited homology to the UL3.5 ORF of PRV (31%). The homolog of this gene is absent in HSV-1. Nucleotide sequence analyses also revealed potential TATA boxes located upstream of each ORF. However, only one polyadenylation signal was detected downstream of the UL3.5 ORF. Northern (RNA) blot analyses revealed four transcripts of 2.4, 1.9, 1.3, and 0.7 kb, which are transcribed in the same direction and are 3'-coterminal transcripts. These mRNAs appear to yield proteins encoded by UL1 (2.4 kb), UL2 (1.9 kb), UL3 (1.3 kb), and UL3.5 (0.7 kb) ORFs. PMID- 7483278 TI - Characterization of DNA binding properties of the immediate-early gene product of equine herpesvirus type 1. AB - The equine herpesvirus type 1 (EHV-1) immediate-early (IE) gene encodes a phosphoprotein that is essential for the activation of transcription from viral early and late promoters and that regulates the transcription from its own promoter. Employment of EHV-1 IE promoter DNA probes and glutathione S transferase fusion proteins harboring truncated portions of the IE gene product in gel shift assays, super shift assays with anti-IE monoclonal antibodies, and DNase I footprinting analyses revealed: (1) amino acid residues 422 to 597 within the 1487-amino-acid IE protein are sufficient for sequence-specific DNA binding; (2) the IE protein binds to EHV-1 DNA at sequences from -11 to +14 that overlap the transcription initiation site (+1); (3) the conserved pentanucleotide 5' ATCGT-3' in the IE promoter located at nucleotides (nt) -6 to -2, relative to the transcription initiation site (+1), is critical for IE protein binding; (4) a weak binding site for the IE protein is also present at nt -92 to -82 of the IE gene within the sequence (-86)ATCGA(-82) in which four of the five nt in the consensus binding sequence are conserved; (5) the IE protein binds to sequences in EHV-1 early and late promoters that contain a degenerate version of the consensus sequence 5'-ATCGT-3'; and (6) mutation of the C or G nt in the pentanucleotide 5'-ATCGT-3' prevents sequence-specific binding of the IE protein, whereas mutation of each of the other three nt only reduces binding. These results suggest that the IE protein can recognize the sites which differ slightly from the proposed consensus sequence. Overall, these findings suggest that formation of a specific complex between an IE protein and its own gene promoter may be a common mechanism used by Alphaherpesvirinae to autoregulate transcription of an essential IE gene. In addition, the finding that the DNA binding domain of the IE protein maps within amino acids 422 to 597, a domain conserved in the IR2 early protein that is a truncated form of the IE protein, suggests that the IR2 protein plays a role in the regulation of the IE gene expression. PMID- 7483279 TI - A C-terminal PreS1 sequence is sufficient to retain hepatitis B virus L protein in 293 cells. AB - Analysis of deletion and/or site-specific mutants of the hepatitis B virus (HBV) env gene, expressed in human cells, provided clues about the mechanism that retains the L protein, the largest gene product, in a pre-Golgi compartment. Differences in secretability of the analyzed variants suggest that the N-terminal myristic acid and an internal sequence within the PreS1 region function as independent retention signals. N-terminal myristic acid alone neither prevented PreS1 + 2 N-linked glycosylation, which signals cotranslational translocation of the domain, nor strongly inhibited lumenal budding. Thus, myristic acid by itself acts by arresting secretion of lumenal, soluble Env particles. By contrast, the internal retention determinant, mapping in the C-terminal portion of PreS1, also prevented budding. In addition, the presence of this PreS1 segment correlated with the depression of PreS1 + 2 glycosylation. This suggests a connection between L retention and the recently described inhibition of PreS1 + 2 cotranslational translocation. A model can be proposed, according to which HBV surface proteins need to cotranslationally translocate their N-terminal moieties in order to assume a transmembrane topology suitable for particulate assembly and secretion. L protein, whose PreS1 + 2 domain undergoes translocation only posttranslationally, would fail to complete the secretion process. To support this model, we show that forced cotranslational translocation of the PreS1 + 2 domain (by attachment of an N-terminal processed signal sequence) results in secretion of L protein. PMID- 7483280 TI - K562 cell strains differ in their response to poliovirus infection. AB - Poliovirus readily establishes a persistent infection in the K562-Mu erythroleukemia cell strain. In this study, three additional K562 cell strains were analyzed for their responses to poliovirus infection and found to be quite variable. K562 cells obtained from the ATCC established a persistent infection, similar to the K562-Mu cell strain, while the majority of cells from two other strains, K562-KI and K562-We, were killed by 4 or 11 days postinfection (p.i.), respectively. Several characteristics of the uninfected and infected cell strains were examined to determine if differences existed which could explain the dramatically different responses to infection. Since K562 cell strains can differentiate toward several cell lineages, the four strains were analyzed for physical and functional likeness to the original K562 cell line using well established functional criteria to determine whether gross changes in differentiation state had occurred. Based on the lack of MHC class I antigen expression and a dose-dependent increase in globin synthesis in response to hemin, all three laboratory K562 cell strains were indistinguishable from the ATCC reference strain. Surface poliovirus receptor levels were also similar in all K562 cell strains, although four- to fivefold lower than those in HeLa cells. Most biochemical events in virus replication either were very similar among K562 cells or were slightly variable and did not correlate with the degree of cell killing. These included levels of virus production, levels of viral protein produced, and processing and turnover of viral polypeptides. The key difference between the cell strains which consistently correlated with cell killing was the degree of virus-induced host translation shutoff, which was always greatest in the most virus-sensitive K562-KI cells. In addition, levels of 2Apro produced in K562 cell strains did not appear to correlate with the levels of host protein shutoff. A related and novel finding in these studies which also strongly correlated with the outcome of infection was the ability of levels of intact p220 to recover by 24 hr p.i. in virus-resistant K562-Mu and -ATCC cells. These data suggest that the key determinants of outcome of infection in this cell model are cytoplasmic host factors related to cytopathology and not factors which may modulate levels of viral protein synthesis or RNA synthesis. PMID- 7483281 TI - Human immunodeficiency virus envelope V1 and V2 regions influence replication efficiency in macrophages by affecting virus spread. AB - The V3 hypervariable region of the HIV-1 envelope protein is a major determinant of viral tropism for macrophages. However, the replication of macrophage-tropic HIV-1 strains varies considerably in macrophages, and this variability has been linked to the V1 and V2 envelope regions. In the present study, recombinant HIV clones were generated by inserting V1 and V2 sequences from the Ba-L HIV isolate, which has a high macrophage replication level, into the genomic background of a macrophage-tropic clone with a low macrophage replication level. Infection of macrophages with varying multiplicities of infection and direct detection of the number of infected macrophages per culture showed that the Ba-L V1 and V2 envelope sequences enhanced the ability of virus to spread in the cultures. In contrast, macrophage-tropic clones with low replication efficiency infected macrophages initially but showed no evidence of spread to additional cells during the culture period. This effect on virus spread appeared to be macrophage specific as it was not observed in cultures of T lymphocytes. Comparison of recombinant clones containing V1, V2, and V3 envelope sequences from high efficiency Ba-L and JR-FL strains indicated that markedly different V1 and V2 sequences could impart the same rapidly spreading phenotype in macrophages. PMID- 7483282 TI - Recovery of virtually full-length HIV-1 provirus of diverse subtypes from primary virus cultures using the polymerase chain reaction. AB - In the course of the global pandemic, the human immunodeficiency virus type-1 (HIV-1) has established at least eight distinct genotypes in the main (M), or prevalent, group of isolates, a variety of rare outlier forms, and intergenotypic recombinants of group M viruses. This genotypic diversity has been documented, for the most part, by sequencing of subgenomic segments of the provirus. Using DNA from virus cultures on peripheral blood mononuclear cells (PBMC) and recent improvements of the PCR technique, we have amplified virtually full-length HIV-1 genomes from genetic subtypes A through G of group M viruses and molecularly cloned several of them. Resequencing of the complete genome of a prototype strain after long PCR amplification and cloning has established a PCR error rate of 0.14%. We also report the first complete PCR-derived sequence of a U.S. clinical isolate of genotype B expanded only in primary PBMC; this provirus harbors a uniquely truncated V3 loop. PMID- 7483284 TI - Molecular biological characterization of the human foamy virus reverse transcriptase and ribonuclease H domains. AB - Foamy viruses form a separate group of retroviruses encoding a pol protein with at least four domains based on comparative sequence alignments. The polymerase and ribonuclease H domains of the human foamy virus (HFV) pol gene were expressed in Escherichia coli either individually or in combination. The histidine-tagged HFV fusion proteins were subsequently purified to near homogeneity by affinity Ni2+ chelate column chromatography. The polymerase and RNase H activities were characterized by performing conventional DNA polymerase and ribonuclease H assays and in situ gel assays. Six purified recombinant HFV proteins were enzymatically active either individually as DNA polymerase and ribonuclease H or as combined domains. The HFV enzymatic activities were characterized with respect to cation preferences and pH optima. Western blots with antibodies against the RNase H domain, in situ reverse transcriptase (RT), and RNase H gel assays showed that in HFV-infected cells pol proteins of 120 and 80 kDa were detectable. A novel activity band of 60 kDa was found in situ RT gel assays. Recombinant RNase H protein additionally purified by fast performance liquid chromatography was capable of removing the primer for minus-strand DNA synthesis when labeled tRNA(Lys1,2) model substrates were used. Specific cleavages occurred at the phosphodiester bonds one to three nucleotides 5' of the RNA-DNA junction. The results revealed biochemical properties of the HFV pol gene products that define functional domains of the HFV pol gene that are distinct but comparable to other retroviruses. PMID- 7483283 TI - Specificity in the association of tomato black ring virus satellite RNA with helper virus. AB - The satellite RNAs (sat-RNAs) associated with some isolates of tomato black ring virus (TBRV) consist of single-stranded molecules of about 1375 nucleotides, encoding a nonstructural protein of 48K which has been shown to be involved in the replication of the sat-RNA. The TBRV sat-RNAs are also dependent for their replication and for their encapsidation on the helper virus. To characterize the nature of the association between sat-RNA and helper virus, transcripts of sat RNA from TBRV isolates C and L (respectively, of serotypes G and S) have been prepared and inoculated onto Chenopodium quinoa leaves or protoplasts. Transcript of the TBRV sat-RNA C is efficiently multiplied when coinoculated with the genomic RNAs of TBRV isolate G (used instead of TBRV isolate C, because isolate G was depleted of sat-RNA), but does not multiply with TBRV isolate L. On the other hand, transcript of the sat-RNA L is able to multiply with the cognate helper virus and, less efficiently, with grapevine chrome mosaic virus (another nepovirus, 80% similar to TBRV), but does not multiply with TBRV G. The specificity of the association resides at the level of sat-RNA replication. Analysis of the multiplication of chimeric sat-RNAs, obtained by exchanging different regions between the two sat-RNAs C and L, showed that the 5' and the 3' noncoding regions of the sat-RNA, although important for replication, are not implicated in specificity. The results suggest that the determinants of the specificity are contained in the 48K sat-RNA-encoded protein. PMID- 7483286 TI - Genotypic characteristics of HTLV-II isolates from Amerindian and non-Indian populations. AB - The Amerindian human T-cell lymphotropic virus type II isolate HTLV-IIG12 has been demonstrated to be an HTLV-IIb with several unique features, including several restriction enzyme site changes, a distinctive pre-gag region, a stop codon within the pol gene, and an extended Tax protein. In this study, HTLV-II isolates from Amerindian and non-Indian populations were characterized by restriction enzyme site analysis to determine the prevalent HTLV-II subtype. In addition, DNA amplification by the polymerase chain reaction and Southern blot analyses were used to probe for the HTLV-IIG12 pre-gag region. Our results showed that of 13 Guaymi Indian isolates subtyped, all were HTLV-IIb, and that approximately one third of 17 isolates had the unique pre-gag region. While other HTLV-II-infected groups contained both HTLV-IIa and HTLV-IIb isolates, none of these isolates showed evidence of the distinctive HTLV-IIG12 pre-gag region. Lastly, DNA sequence analysis was used to determine the prevalence of the stop codon within the pol gene open reading frame. These analyses revealed that the occurrence of a stop codon within this sequence appeared to be characteristic of most HTLV-IIb subtypes. These results further our understanding of the genetic variations and evolution of the HTLV-II viruses within the endemically infected Amerindian populations, as well as U.S. intravenous drug users and other non Indian populations. PMID- 7483285 TI - Biologic activity of human papillomavirus type 16 E6/E7 cDNA clones isolated from SiHa cervical carcinoma cell line. AB - Three species of E6/E7 cDNAs of human papillomavirus type 16 (HPV16) for the full length E6/E7 and spliced E6*I/E7 and E6*II/E7 mRNAs were synthesized by reverse transcriptase-(RT-)PCR from RNA of the cervical carcinoma cell line SiHa. Two cDNA mutants carrying point mutations in either a splice donor site or acceptor site within the E6 open reading frame were also constructed. These HPV16 E6/E7 cDNAs were cloned under the SV40 enhancer/promoter and the MMTV LTR to examine the activities of ras-collaborative transformation and induction of cellular DNA synthesis, both of which depend on the E7 gene product. The E6*II/E7 cDNA and two mutated cDNAs deficient in the spliced mRNA transcription showed lower levels of both activities than the full-length E6/E7 and the E6*I/E7 cDNA. The rat cell lines carrying each of the E6/E7 cDNAs contained the E6/E7 mRNA species expected. A small amount of E6*I/E7-sized mRNA was transcribed from a splice-donor site mutant of the E6/E7 cDNA, which turned out to be a transcript derived from a cryptic splice donor site six bases upstream from the conventional site. Among NIH3T3 cells carrying one of the above-mentioned E6/E7 cDNAs, the cells expressing E6*I/E7 mRNA [cells carrying cF(wt) and c*I] produced an amount of E7 protein comparable with those carrying the E7 or E6E7 region. These results suggest that the E6*I/E7 is the mRNA that is important for the efficient expression of E7 product from the HPV16 E6/E7 region. PMID- 7483287 TI - Sequence and phylogenetic analysis of the VP1 gene in two cell culture-adapted HAV strains from a unique pathogenic isolate. AB - The nucleotide sequences of the VP1 coding region of two newly characterized, cell culture-adapted hepatitis A virus (HAV) strains (RG-SB11 and RG-SB16) were analyzed and compared with homologous regions of previously characterized HAV strains of human or monkey origin, and at different levels of tissue-culture adaptation. In particular, HM175wt and its derivative strains and MBB, LCDC1, PA21, and AGM27 isolates were considered. RG-SB11 and RG-SB16 HAV strains were derived from a pathogenic isolate from an acutely infected patient, purified from stool, and subjected to different strategies of adaptation. Several nucleotide differences were observed, but high conservation was found in the predicted VP1 protein sequences, which confirms structural constraints for this region. Furthermore, comparative amino-acid sequence analysis of VP1 from all HAV isolates studied has shown, particularly for those from naturally infected monkeys, that differences are limited to the amino and carboxy-terminal part of the molecule. The results of phylogenetic analysis have confirmed the common origin of the RG-SB11 and RG-SB16 strains. The complete nucleotide sequences of the VP1 coding region of the RG-SB11/16, HM175 derivative strains and of other HAV strains has shown that branch-length evolution can give a measure of the evolution of HAV during adaptation processes. PMID- 7483288 TI - Quantitative PCR as a method for monitoring retroviral infection on the gene level. AB - For monitoring retroviral infection on the gene level, we propose the use of quantitative PCR with two internal standards: one for a fragment of the viral genome and the other for the host cell marker gene. The standards (one for HIV and the other for a human DNA marker gene HLA-DQ alpha) were constructed by PCR mediated joining of DNA fragments and were found to be effective in quantitative PCR despite rather different structures of amplified fragments in target and standard DNAs. The number of HIV DNA copies was found to be 2-500 per 1000 lymphocytes in blood from HIV-infected patients and up to 5000+ per 1000 cells in chronically infected cell lines. The degree of infection thus measured was found to change over the course of treatment. PMID- 7483289 TI - Foot and mouth disease virus replication in bovine skin Langerhans cells under in vitro conditions detected by RT-PCR. AB - The replication of foot and mouth disease virus (FMDV) was studied in isolated bovine skin Langerhans cells (LC), in keratinocytes from epidermal cell suspension, and in migrating LC obtained from cultured bovine epidermal sheets in vitro. Viral RNA replication in infected cells was determined by the reverse transcriptase-polymerase chain reaction (RT-PCR) of the negative FMDV RNA strand and by the plaque forming assay of FMDV. It was established that bovine skin LC, keratinocytes, and migratory bovine LC infected with FMDV strain 01 Geshur supported virus replication. This RT-PCR method to detect the negative strand of FMDV RNA in migratory bovine skin LC may be useful for determining FMD virus replication in tissue cells. PMID- 7483291 TI - Characterization of a conserved gene block in the murine cytomegalovirus genome. AB - The nucleotide sequence between the lytic origin and the gH gene of the murine cytomegalovirus genome (map units 0.416-0.455) was determined. Five of the 15 open reading frames identified encode polypeptides, which exhibit significant homology to polypeptides of human cytomegalovirus (HCMV) (UL69, UL71, and UL73) and to polypeptides of several other herpesviruses (dUTPase and helicase/primase). The location in the genome of the five open reading frames and their direction of transcription is perfectly conserved between murine cytomegalovirus (MCMV) and HCMV. These data suggest that MCMV and HCMV have a highly related genome organization. Thus, MCMV offers a good model for molecular and functional analysis of cytomegalovirus genes in vitro and in vivo. PMID- 7483292 TI - Nucleotide sequence of the thymidine kinase gene of raccoon poxvirus. AB - Using primers directed against the thymidine kinase gene of vaccinia virus, PCR products were generated from nucleic acids extracted from raccoon poxvirus infected Vero cells. The PCR products were consistent in size with the expected products from vaccinia virus. Nucleotide sequence determination revealed that the raccoon pox thymidine kinase gene and flanking regions were 84.3% homologous to the corresponding sequences of vaccinia virus. At the amino acid level, an open reading frame coding for a polypeptide of 177 amino acids was found with 87% homology to thymidine kinase of vaccinia. PMID- 7483294 TI - Changes in the NS gene of neurovirulent strains of influenza affect splicing. AB - The nonstructural (NS) gene has been identified as an accessory factor in determining neurovirulence for influenza A virus. The nucleotide sequence of the NS gene of the neurovirulent variant A/NWS/33 was determined and compared with its parent, A/WS/33. Alterations in the mRNA structure of the gene were observed that serve to mask the 3' splice site. Changes in this region were shown to correlate with reduced splicing of the NS gene in neurovirulent strains. PMID- 7483293 TI - HTLV-I from Iranian Mashhadi Jews in Israel is phylogenetically related to that of Japan, India, and South America rather than to that of Africa and Melanesia. AB - A new endemic focus of human T-lymphotropic virus type I (HTLV-I) was recently reported among Mashhadi Jews, a group of immigrants from northeastern Iran to Israel. We extracted DNAs from fresh peripheral blood mononuclear cells (PBMCs) and/or gargle mouthwash from 10 HTLV-I carriers, who consisted of members of one family, and HTLV-I-associated myelopathy (HAM) and adult T-cell leukemia (ATL) patients. Long terminal repeat (LTR) regions of proviral DNAs were sequenced and analyzed phylogenetically. In a phylogenetic tree, all the Mashhadi HTLV-I isolates belonged to subtype A, one of the three subtypes of the cosmopolitan type of HTLV-I, and made a tight cluster distinct from the other isolates of subtype A from Japan, India, the Caribbean Basin, and South America. Although a few nucleotide substitutions were observed among the clones sequenced, no characteristic sequence variation was found in different disease manifestations, even in one family or different sources of DNA preparation. PMID- 7483290 TI - Functional organization of variola major and vaccinia virus genomes. AB - Comparison of the genomic organization of variola and vaccinia viruses has been carried out. Molecular factors of virulence of these viruses is the focus of this review. Possible roles of the genes of soluble cytokine receptors, complement control proteins, factors of virus replication, and dissemination in vivo for variola virus pathogenesis are discussed. The existence of "buffer" genes in the vaccinia virus genome is proposed. PMID- 7483295 TI - Nucleotide sequences of the neuraminidase genes of influenza A/Leningrad/134/57 (H2N2) virus and two of its live, attenuated, cold-adapted variants. AB - The nucleotide sequences of the neuraminidase (NA) genes of the A/Leningrad/134/57 (H2N2) wild-type (Len/wt) virus as well as two of its live attenuated, cold-adapted (ca) variants, A/Leningrad/134/17/57 (Len/17) and A/Leningrad/134/47/57 (Len/47), were determined. In comparison with Len/wt, one nucleotide change (C-225 to A) was found in the NA gene of Len/17. This change codes for a Thr-to-Asn substitution at position 69 of NA. The NA gene of the more attenuated Len/47 ca virus has one silent (T-814 to C) and two coding nucleotide substitutions, C-78 to T (Ala-20 to Val) and C-225 to A (Thr-69 to Asn). These sequence data were used to design a PCR-restriction technique to determine the origin of the NA gene in candidate live, attenuated vaccine reassortants made by reassorting these ca strains with current field viruses. PMID- 7483296 TI - HBx protein of hepatitis B virus interacts with the C-terminal portion of a novel human proteasome alpha-subunit. AB - Two-hybrid protein interaction screening in yeast was used to identify proteins that interact with the HBx nonstructural protein of hepatitis B virus (HBV). A new human member of the proteasome alpha-subunit family was obtained. Its protein sequence closely resembles the 28 kD subunits from other organisms. The interaction with HBx was abolished by a two amino-acid insertion behind position 128 in HBx, in a region previously found to be essential for its transcriptional transactivation function. These data support a model of HBx acting indirectly on transcriptional processes. By binding to a specific proteasome alpha-subunit, HBx might interfere with degradative processes, thereby enhancing the half-life of different transcription factors and other nuclear regulatory proteins. Interaction with the Hu 28k proteasome subunit could thus provide a unifying explanation for the markedly pleiotropic effects of HBx. PMID- 7483297 TI - Continuous gradient index and shell models of the human lens. AB - Because of the complexity of tracing rays through a gradient refractive index medium, the human eye's lens is sometimes approximated by a shell structure with constant refractive index within shells. In the shell model, power arises from a combination of an axial variation in index and the curvatures of the shell surfaces. We develop an equation which gives the power due to the gradient index of the lens, and use this to choose shell models that have the same power as the continuous gradient index model. Some types of shell models are described and evaluated. PMID- 7483298 TI - Sharpening of drifting, blurred images. AB - The perceived blur of moving images is less than expected given the sluggish temporal response of the visual system. This suggests that a motion deblurring mechanism may exist to preserve the positional acuity and sharpness of moving images. Furthermore, when sequences of blurred stills are presented, observers report that the moving image is in sharp focus raising the possibility that there is a mechanism which may sharpen the appearance of moving, blurred images. We have measured the effects of velocity and contrast on the perceived blur of drifting, blurred images (sine gratings and blurred edges). Subjects matched the perceived blur of drifting, blurred images to that of static, blurred images in a dimly lit room. It was found that perceived blur was inversely related to drift speed and contrast. The results confirm that moving, blurred images may appear sharper than when they are static. This finding is not consistent with some models of motion deblurring since these account only for the preservation of sharp contours that are present in the image and not for the sharp appearance of images that are in fact blurred. PMID- 7483299 TI - Mechanisms of purely subjective contour tilt aftereffects. AB - Neurones tuned for second-order stimuli--those which have edges defined by properties other than luminance and colour--have been frequently observed in prestriate cortex and in area V2 there are neurones which explicitly and unambiguously signal the orientation of purely subjective contours, i.e. contours with no Fourier components at the orientation of the perceived edge [von der Heydt, R. & Peterhans, (1989) Journal of Neuroscience, 9, 1731-1748]. No neurones in area V1 showed similar tuning characteristics. In addition, it has been demonstrated that like real contours, purely subjective test contours are subject to tilt aftereffects following prolonged viewing of an adapting stimulus. whether that stimulus is real or subjective [Paradiso, M. A., Shimojo, S. & Nakayama, K. (1989) Vision Research, 29, 1205-1213]. This result supports the assertion that the cortical processes responsible for real contour perception are also those giving rise to subjective contour perception. The data reported here further examined this hypothesis. Four experiments show that purely subjective contours exhibit both direct and indirect tilt aftereffects and tilt illusions like those observed with real contours. Further, they provide evidence that direct and indirect subjective contour effects, like direct and indirect real contour effects, arise via the operation of two mechanisms: a low level process, possibly lateral inhibition between orientation channels, and a second "higher-order" process. The data suggest that processing of orientation information beyond the striate cortex is similar to that which occurs in area V1 and the data are consistent with models of contour processing which assume that all perceived contours, both real and subjective, arise from a common mechanism. PMID- 7483300 TI - Binocular combination of contrast signals from orthogonal orientation channels. AB - In a series of psychophysical experiments, observers compared the apparent contrast of a dichoptic stimulus (contrast edges of orthogonal orientations in the two eyes) and a monocular stimulus ( a pie pattern in one eye and a blank field in the other eye). When the contrast of the dichoptic stimulus was low (2 3%) the two orthogonal edges were perceived as a stable pie pattern and its apparent contrast matched with that of a monocular pie pattern whose contrast was twice the contrast of the dichoptic stimulus. The dichoptic contrast gain (monocular/dichoptic contrast ratio) decreased with increasing dichoptic contrast. Although the observers showed an inability to discriminate the apparent contrasts of the dichoptic and the monocular pie patterns, they were able to tell which stimulus was the dichoptic one in a forced choice paradigm. A binocular contrast preservation model was proposed for binocular combination of contrast signals from orthogonal orientation channels. PMID- 7483301 TI - Spectral characteristics of blink suppression in normal observers. AB - Previous studies of the characteristics of suppression occurring under various visual conditions show similarities and differences which may indicative of the mechanism of suppression. The primary purpose of this study was to determine if the suppression that occurs in response to an eyelid blink (blink suppression) is similar to that which occurs during a saccade (saccadic suppression). In addition, the characteristics of blink suppression and other forms of suppression (i.e. permanent and binocular rivalry suppression) are compared. A test probe paradigm was utilized to determine the effect of blink suppression on the spectral sensitivity function in three normal observers. Employing a two alternative forced choice technique, thresholds were determined for wavelengths from 420 to 680 nm in 20 nm steps. At each wavelength, the threshold was determined at 0 and 400 msec after the onset of a voluntary blink. The magnitude of suppression was taken as the difference between the 0 and 400 msec thresholds. Similar to saccadic suppression, the magnitude of blink suppression increased as the stimuli biased detection towards the luminance channel. These results suggest that blink suppression and saccadic suppression are the result of a single mechanism. Similarities between blink suppression and other forms of visual suppression are also considered. PMID- 7483302 TI - Variation in color matching and discrimination among deuteranomalous trichromats: theoretical implications of small differences in photopigments. AB - Individual differences in abnormal color vision are well known. A fundamental unresolved problem is the great variation in color vision even among those classified as having the same color-vision defect. Several physiological hypotheses have been proposed to account for this variation but little consideration has been given to how (and how much) color matching and discrimination are affected by the posited physiological mechanisms. Advances in molecular genetics have renewed interest in this problem, which is at the foundation of the relation between genotype and phenotype. We report here theoretical Rayleigh ranges (chromatic discrimination) and quantal matches for deuteranomalous trichromats with photopigments in the red/green range that vary in their separation and optical density. The results show there is relatively little loss of discrimination with pigments of normal optical density separated by as little as 2-3 nm. With pigments separated by 4 nm or less, however, optical density can strongly influence discrimination when varied independently in the two types of cone. Moderately lower (or higher) optical density in only one cone type affects discrimination by altering the shape of the cone's relative spectral sensitivity function. The lack of correlation between Rayleigh-match midpoint and range, which is reported in the literature, may be accounted for by independent variation in pigment separation and optical density. PMID- 7483303 TI - Global motion perception: no interaction between the first- and second-order motion pathways. AB - The experiments reported here address the issue of whether the pathways which extract motion from first-order and second-order spatial patterns remain separate or whether they combine at some higher level in the motion system to form a single pathway. The question is addressed by investigating the interaction of first-order and second-order stimuli in the processing of a global-motion stimulus [a variant of the task introduced by Newsome & Pare (Journal of Neuroscience, 8, 2201-2211, (1988)]. Two experimental procedures were used. The first consisted of determining the effect of the addition of dots of one type (e.g. first order) undergoing purely random motion on the ability to extract the global-motion signal carried by dots of the other type (e.g. second order). The second experimental procedure consisted of determining the effect of maintaining a coherent-motion signal in one type of dot, moving in the opposite direction to the global-motion direction, on the ability to extract the global-motion signal carried by dots of the other type. The dots were matched for their effectiveness in producing a global motion percept and the results for both procedures were the same. First-order dots impaired the ability to extract second-order global motion, and second-order dots had no effect on first-order global-motion extraction. It is argued that the sensitivity of the second-order global-motion system to the first-order dots is due to the ability of the second-order local motion detectors to detect these dots. The present results are thus interpreted as indicating that the first-order and second-order motion pathways remain separate up to and including the level in the motion system at which global motion signals are extracted. PMID- 7483304 TI - The critical role of relative luminance relations in White's effect and grating induction. AB - It has been proposed that both White's effect and the grating induction effect are examples of brightness induction phenomena modeled in terms of local spatial filters. We have shown that for these illusions to occur it is necessary that the luminance of the gray target elements falls between that of the inducing stripes of the square-wave pattern. This critical role of luminance relationships is not predicted by existing models of these illusions. PMID- 7483305 TI - Intertrial effects of randomization on saccadic reaction times in human observers. AB - We investigated the effect of randomizing different spatial and temporal parameters on saccadic reaction times (SRTs) in five human subjects, to explore the relative occurrence of express and regular saccades. Parameters randomized in various test sessions were: target direction (right/left), intertrial interval, fixation foreperiod and gap duration (two or three 400 msec) in gap and overlap trials. For the sake of comparison the parameters under consideration were kept constant in non-random control sessions. We found that express saccades were still present in the random test sessions but their relative frequency (and the number of regular saccades) obtained in a given test session depends on the type of randomized parameters: randomizing the intertrial interval or the fixation foreperiod in the gap task yielded modest but significant changes in the SRT distributions, express and fast regular saccades being present in both the control and the random conditions. Randomization of the fixation foreperiod in the overlap task, on the other hand, caused a quite drastic increase in the SRTs. Randomization of gap and overlap trials did not cause considerable effects on express and fast regular saccades in the gap trials, and the SRTs in the overlap trials were significantly increased only in two subjects. When two or three gap durations were randomly interleaved, we found effects that ranged from "negligible" (usually for the longest gap in a given test session) to highly significant differences as compared with the corresponding control condition. The results suggest that express saccades--as fast regular saccades--are visually guided saccades which occur when a certain state of saccade preparation has been reached before target onset. This state depends on the amount of activation in the brain structures involved in the control of attention and fixation, and the decision processes involved in saccade preparation. PMID- 7483306 TI - A Bayesian framework for understanding texture segmentation in the primary visual cortex. AB - This paper presents a mathematical theory for understanding the computations involved in texture segmentation in the primary visual cortex. We propose that texture segmentation is a part of the early visual system's overall strategy to infer surfaces of objects in a visual scene. Based on this insight, we use the Bayesian inference paradigm to formulate the texture segmentation problem into a maximum a posteriori surface inference problem. The dynamical system for finding the optimal solution of this problem can be characterized by two concurrent and interactive processes: a gradual sharpening of the boundary signals and a simultaneous smoothing of the surface signals. The behavior of these dynamical processes was studied using both analytical and computational methods. We present some computational results and mathematical predictions. This theory suggests a novel framework for understanding the functional roles of the complex cells in the primary visual cortex. PMID- 7483307 TI - Increasing doses of l-sulpiride reveal dose- and spatial frequency-dependent effects of D2 selective blockade in the human electroretinogram. AB - The amplitude and phase of the second harmonic (15 Hz) of the electroretinographic responses to three different spatial frequency grating stimuli (0.25, 1 and 4 c/deg), reversed at 7.5 Hz, were studied i normal human subjects, before and 30 min after the systemic administration of three doses (0.071, 0.357 or 1.428 mg/kg) of a selective D2 blocker, l-sulpiride, to three populations of 18, 19, or 20 subjects. The effect of the drug on the pattern electroretinogram (PERG) was clearly dose-dependent, being greatest on the responses to 4 c/deg. The mean decrease in second harmonic amplitude was -13.8% after 0.071 mg/kg of l-sulpiride, -23.5% after 0.357 mg/kg and -28.5% after 1.428 mg/kg. The last two variations were significant at P < 0.01 and P < 0.01 respectively. These data suggest that a dose-dependent effect on the human retinal response to 4 c/deg stimuli exists, probably mediated by a coupling between l-sulpiride and D2 receptors. Lastly, our data suggest that D2 receptors may play an important role in the pathophysiology of visual dysfunction in Parkinson's disease, that has been described to be more significant at medium spatial frequency (2-5 c/deg). PMID- 7483308 TI - Different effects of dopamine and piribedil (a dopamine D2 agonist) on frog monocular optokinetic nystagmus asymmetry. AB - Frog monocular optokinetic nystagmus (OKN) displays a directional asymmetry, reacting only to stimulations in the temporal-nasal (T-N) direction. The nasal temporal (N-T) component is almost absent. The systemic or intrapretectal injection of Piribedil, a D2 dopamine agonist, provokes the appearance of a N-T component suppressing the monocular OKN asymmetry. Conversely, dopamine or haloperidol (a dopamine antagonist, acting mainly on D2 receptors) have no effect upon the monocular OKN unidirectionality. The monocular OKN N-T component still appears after administration of Piribedil even if this injection is preceded by administration of haloperidol which blocks the dopaminergic D2 receptors. Moreover administration of atropine (a cholinergic muscarinic antagonist) following that of Piribedil suppresses the N-T component; when injected before Piribedil, atropine prevents the appearance of the N-T component. These results suggest that in our experiments, Piribedil binds with muscarinic receptors. PMID- 7483309 TI - Reversals of the colour-depth illusion explained by ocular chromatic aberration. AB - Although many colour-depth phenomena are predictable from the interocular difference in monocular chromatic diplopia caused by the eye's transverse chromatic aberration (TCA), several reports in the literature suggest that other factors may also be involved. To test the adequacy of the optical model under a variety of conditions, we have determined experimentally the effects of background colour on perceived monocular chromatic diplopia and perceived depth (chromostereopsis). A Macintosh colour monitor was used to present red, blue, and green test stimuli which were viewed monocularly or binocularly (haploscopically) through 1.78 mm artificial pupils. These apertures were displaced nasally and temporally from the visual axis under controlled conditions to induce a variable degree of TCA. Monocular chromatic diplopia and binocular chromostereopsis were measured for red and blue targets, and also for red and green targets, presented on either a black background or on a background which was composed of the sum of the targets' spectral composition (e.g. red and blue presented on magenta; red and green presented on yellow). In all cases, chromatic diplopia and chromostereopsis were found experimentally to reverse in sign with this change in background. Furthermore, we found that a given coloured target could be located in different depth planes within the same display when located on different background colours. These seemingly paradoxical results could nevertheless be explained by a simple model of optical TCA without the need to postulate additional factors or mechanisms. PMID- 7483310 TI - Discrimination of compound gratings: spatial-frequency channels or local features? AB - Models based on spatial-frequency channels and local features provide alternative explanations for suprathreshold pattern discrimination. We compared psychophysical discrimination data with the predictions of the Wilson and Gelb channel model and three local-feature models. The features were peak-valley local contrast, peak-peak local contrast, and luminance gradients. We measured visual sensitivity for discriminating compound gratings (F + 3F or F + 5F, in peaks-add or peaks-subtract phases) whose component contrasts were yoked together so that a contrast increment in one component was accompanied by an equal decrement in the other. The Wilson and Gelb model accounted for the results with peaks-add gratings, but failed to predict those with peaks-subtract gratings. None of the local-feature models explained the results by themselves. Most of the data fell close to an envelope composed of the lowest thresholds of the three feature detector models, although there were important exceptions. Our findings are consistent with the view that suprathreshold pattern discrimination is mediated by mechanisms responsive to spatially localized features and that more than one type of feature is used. PMID- 7483311 TI - The functional architecture of human visual motion perception. AB - A powerful paradigm (the pedestal-plus-test display) is combined with several subsidiary paradigms (interocular presentation, stimulus superpositions with varying phases, and attentional manipulations) to determine the functional architecture of visual motion perception: i.e. the nature of the various mechanisms of motion perception and their relations to each other. Three systems are isolated: a first-order system that uses a primitive motion energy computation to extract motion from moving luminance modulations; a second-order system that uses motion energy to extract motion from moving texture-contrast modulations; and a third-order system that tracks features. Pedestal displays exclude feature-tracking and thereby yield pure measures of the first- and second order systems which are found to be exclusively monocular. Interocular displays exclude the first- and second-order systems and thereby to yield pure measures of feature-tracking. RESULTS: both first- and second-order systems are fast (with temporal frequency cutoff at 12 Hz) and sensitive. Feature tracking operates interocularly almost as well as monocularly. It is slower (cutoff frequency is 3 Hz) and it requires much more stimulus contrast than the first- and second-order systems. Feature tracking is both bottom-up (it computes motion from luminance modulation, texture-contrast modulation, depth modulation, motion modulation, flicker modulation, and from other types of stimuli) and top-down--e.g. attentional instructions can determine the direction of perceived motion. PMID- 7483312 TI - Bayesian analysis of identification performance in monkey visual cortex: nonlinear mechanisms and stimulus certainty. AB - The identification performance of single neurons in the primary visual cortex was quantified by measuring how accurately one could know the stimulus based upon the neuron's response. We found that for a typical neuron a response of 10 action potentials, following one brief stimulus presentation, was sufficient to classify the stimulus as belonging to a relatively small region in stimulus space, with a high degree of confidence. The performance was better than that which could be attained through linear summation of excitation and inhibition alone. The results suggest that the enhanced performance is a consequence of two nonlinear mechanisms: contrast gain control and expansive response exponent. PMID- 7483313 TI - Natural problems for stereoscopic depth perception in virtual environments. AB - The use of virtual reality (VR) display systems has escalated over the last 5 yr and may have consequences for those working within vision research. This paper provides a brief review of the literature pertaining to the representation of depth in stereoscopic VR displays. Specific attention is paid to the response of the accommodation system with its cross-links to vergence eye movements, and to the spatial errors that arise when portraying three-dimensional space on a two dimensional window. It is suggested that these factors prevent large depth intervals of three-dimensional visual space being rendered with integrity through dual two-dimensional arrays. PMID- 7483314 TI - Spatial frequency channels in experimentally strabismic monkeys revealed by oblique masking. AB - Although the spatial vision deficits of human strabismic amblyopes have been well documented, surprisingly little is known about the mechanisms underlying their visual performance. In an effort to reveal the structure underlying the spatial vision deficits associated with strabismic amblyopia, we measured the performance of monkeys (Macaca nemestrina) with experimental strabismus in a contrast detection task with oblique masks. The masks were two adjacent identical oblique sine-wave gratings modulated in space by a Gaussian envelope. The target stimulus was a vertically oriented Gabor patch that appeared superimposed on the center of either the left or the right mask. The animals were trained by operant methods to indicate the location of the target. We measured detection thresholds in each eye independently for a large number of test and mask spatial frequencies. For each test spatial frequency, detection thresholds were elevated in the presence of the mask. The threshold evaluations showed a peak for a particular spatial frequency that was typically similar to the test spatial frequency. This pattern of results is consistent with the idea that the tests are detected by a discrete number of channels tuned to a narrow range of spatial frequencies. The data from the deviated eyes did not appear qualitatively different from those of the fellow eyes, and could be accounted by the same number of channels in both eyes. Quantitative estimates of the channels' characteristics revealed that the channels derived from the deviated eyes' data were similar to those yielded by the fellow eyes, but showed a reduction in their sensitivity to contrast. PMID- 7483315 TI - The spatial precision of macaque ganglion cell responses in relation to vernier acuity of human observers. AB - Responses of parafoveal macaque ganglion cells were measured as a function of the contrast and position of an edge flashed within their receptive fields. The goal was to determine the ability of different cell types to signal edge location. For comparison, parafoveal vernier thresholds of human observers were measured with pairs of flashed edges. Cells of the magnocellular (MC-) pathway gave larger responses than cells of the parvocellular (PC-) pathway. Neurometric analyses comparing a cell's response at different edge positions were performed. The positional signal from single MC-pathway cells was more precise than from PC pathway cells, especially at lower contrasts. In a second analysis, based on the neurophysiological results, responses from a matrix of ganglion cells were generated. Using a simple model, vernier performance expected from such a matrix was predicted as a function of edge length and contrast. Again, the MC-pathway gave a more precise positional signal than the PC-pathway despite the latter's numerical advantage. At contrasts of 20% and below, only the MC-pathway would appear capable of supporting vernier performance with our stimuli. At higher contrasts either the MC- or PC-pathway could provide an adequate signal. PMID- 7483316 TI - The perceived orientation of aliased lines. AB - The use of raster display devices for the display of graphics causes problems of aliasing when edges or lines are produced. This can be significant in those psychophysical experiments where the orientational properties of the stimulus are important. We have assessed the perceived orientation of a selection of aliased lines by comparing them with the orientation of pairs of dots. It is found that the perceptual orientation is modelled well by a least-squares metric on the pixels that compose the line. Small deviations from this metric were found, and were also found in a control experiment employing anti-aliased lines. They appear to be due to range effects. Averaged across subjects, orientational acuity was only slightly lower for aliased lines. PMID- 7483317 TI - The effect of nearby luminance contrast polarity on color boundary localization. AB - The study measured the shift in apparent position of a color-defined target boundary as a function of the distance, luminance polarity and amount of contrast of a nearby luminance-defined flanking boundary. In general, the position of the target boundary shifted towards the flank with the attraction being somewhat greater for negative than positive polarity flanks, and for high compared to low contrast flanks. High contrast, negative polarity flanks resulted in greater attraction at 3.69 min arc separation. For low contrast flanks, the apparent shift in position of the target boundary depended on the polarity and position of the flank relative to the target. For example, for small separations ( < 3 min arc) flank polarity had little influence, while for larger separations ( > or = 3.69 min arc), negative polarity flanks exhibited attraction while positive polarity flanks began to show repulsion. The results support the notion that luminance and color processing may share a common representation for the localization of boundaries. Position judgments based on this representation appear to be influenced by the amount of luminance contrast in a nearby boundary. PMID- 7483318 TI - Absence of adaptive plasticity after voluntary vergence and accommodation. AB - Subjects maintained their eyes crossed (verged) for a period of 8 min in darkness with monitoring provided by an infrared video system. Changes in resting vergence (RV) and resting focus (RF) were examined. Results showed: (i) visual stimulation was not necessary for adaptation of either RV or RF, but (ii) these purely motor effects were significantly smaller and more dissipative than those attributable to visually driven adaptation, and (iii) voluntary vergence amplitude was negatively correlated with pupil size. Assuming that voluntary vergence is driven by accommodation, then the voluntary signal must enter the oculomotor control system prior to the cross links between channels, but beyond the site of the visually driven adaptive elements. PMID- 7483319 TI - The nature of the inputs to cortical motion detectors. AB - Recently, Jagadeesh, Wheat and Ferster [(1993) Science, 262, 1901-1904] presented intracellular recordings from direction-selective simple cells in primary visual cortex and provided an analysis to support the idea that synaptic summation in simple cells is linear. New analysis presented in this study reveals that: (1) the number of subunits contributing to the analyzed simple cell inputs is two; (2) the subunits are nonlinear in the time domain; (3) each subunit linearly integrates the luminance across the receptive field being, thus, linear to local contrast; (4) the waveforms of the subunit signals are linearly modulated by local contrast at the subunit loci unless the contrast changes its sign; (5) the synaptic summation in the simple cell is linear; (6) nonlinearity of even harmonics has sufficient information for retrieving of relative spatial phase of the subunits and reconstruction of the exact temporal profiles of the subunit signals. PMID- 7483320 TI - Effect of repeat use and coating defects of gold foil electrodes on electroretinogram recording. AB - Qualitative defects and resistances in 94 used gold foil electrodes were examined. The electrodes were divided into four groups of varying resistances and gold coating defects. Ten were randomly selected from each group to measure standard photopic electroretinograms (ERGs) in a normal subject. Ten new electrodes were used as controls. There was no significant difference among the electrode groups for ERG peak implicit times or amplitudes, although a slightly greater amplitude variability was observed for the groups with more defects. Provided the resistance of the electrode is low ( < 5 omega), its reuse at least for ganzfeld ERGs appears warranted regardless of apparent coating defects. PMID- 7483321 TI - cAMP-dependent regulation of gene transcription by cAMP response element-binding protein and cAMP response element modulator. PMID- 7483322 TI - Regulation of G protein-coupled receptors by receptor kinases and arrestins. PMID- 7483323 TI - Vasopressin and oxytocin: molecular biology and evolution of the peptide hormones and their receptors. PMID- 7483324 TI - Structure and function of estrogen receptors. PMID- 7483325 TI - Phosphorylation and steroid hormone action. PMID- 7483326 TI - Nucleocytoplasmic shuttling of steroid receptors. PMID- 7483327 TI - Transcriptional regulation of the genes encoding the cytochrome P-450 steroid hydroxylases. AB - Steroid hormone biosynthesis requires the concerted action of a related group of cytochrome P-450 steroid hydroxylases. In recent years considerable effort has been directed toward defining the molecular basis for the cell-selective expression of these genes and their transcriptional regulation by trophic hormones. The orphan nuclear receptor SF-1, acting through a conserved element found in the proximal promoter regions of all steroid hydroxylase genes, seems to be a major, but not exclusive, determinant of cell-selective gene expression. In contrast, the coordinate responses of the steroid hydroxylases to trophic hormones apparently involves an interplay of multiple proteins that collectively lead to a synchronous induction of gene expression. In some instances these interactions apparently involve transcription factors that also contribute to the cell-selective expression of these genes. PMID- 7483328 TI - Stress and the brain: a paradoxical role for adrenal steroids. PMID- 7483329 TI - Retinoids and mouse embryonic development. PMID- 7483330 TI - Multiple facets of the modulation of growth by cAMP. PMID- 7483331 TI - [25 years' experience with diagnosis of pheochromocytoma]. AB - The author submits his 25-year experience with the diagnosis of pheochromocytoma in 36 patients. He evaluates the clinical picture by discrimination analysis of subjective symptoms and objective signs of the disease. Hypertension--by 24-hour monitoring of the blood pressure. In case of slight elevation of catecholamines he uses the Estulice test. In liminal cases patients are dispensarised. Due to a higher frequency of CT examinations in hypertonic patients findings of adenomas of the adrenals cumulate which are hormonally inactive. Gammagraphy can in that case confirm a functionally active pheochromocytoma, it can however produce also a falsely positive finding with an extraadrenal localization. Non-invasive detection of the site of a pheochromocytoma (computed tomography, ultrasonography, metaiodobenzyl guanidine gammagraphy, the finding of an increased catecholamine production) suffices for indication of surgical treatment. The author tries to avoid invasive examinations (angiography), as well as pharmacological provocation tests. Symptoms of catecholamine heart muscle disease can simulate the electrocardiographic finding of acute myocardial infarction which makes decision taking on the therapeutic procedure more difficult. After alpha-blockers and in particular after a successful operation the electrocardiographic changes are restored to normal. After early assessment of the diagnosis and surgery the disease has at present a favourable prognosis. PMID- 7483332 TI - [Diagnostic sensitivity of 24-hour electrocardiographic Holter monitoring in indications for coronarography]. AB - The authors evaluated 24-hour ECG monitoring (Holter monitoring) in 70 patients indicated for coronarography. They assessed the sensitivity, specificity, positive and negative predictive value of Holter monitoring. They compared the findings with data reported in the literature and with the sensitivity of the ergometric test for indication of patients for coronarographic examination. Although the sensitivity and specificity of Holter monitoring does not exceed generally reported values of sensitivity and specificity of bicycle ergometry, in the discussion the authors mention conditions where Holter monitoring is an asset for indication of patients for coronarographic examination. PMID- 7483333 TI - [Hepatitis C virus antibodies in hemodialyzed patients]. AB - Assessment of antibodies against the hepatitis C virus (HCV) in 629 patients treated by haemodialysis in 47 dialyzation centres revealed the probability of HCV infection in 196 of them, i.e. in 31%. In a group of 91 patients newly admitted for dialysis, antibodies against HCV were found in 24, i.e. in 26%. The percentage of anti-HCV antibody carriers in different dialyzation units varied between 6 nad 60. PMID- 7483334 TI - [A relapsing and protracted form of viral hepatitis A: comparison of adults and children]. AB - In recent years in children and adults with acute viral hepatitis A relapses and a protracted course of the disease were described. The authors followed up 37 patients with viral hepatitis A (20 children and 17 adults) and compared the clinical course of the disease, the period of hospitalization, persistence of IgM anti-HAV antibody in serum, the incidence of relapses and protracted forms of the disease. In adults the mean hospitalization period was longer (28 days as compared with 19), the average serum bilirubin value was higher (94 mumol/l as compared with 51 mumol/l), there were more cases with obvious jaundice (59% as compared with 30%) and the early serum antibody IgM anti-HAV persisted longer (19 weeks as compared with 14 weeks). Relapses of the disease were equally frequent (12% vs. 10%), however adults had more often a protracted course of hepatitis (23.5% vs. 10%). The observed differences were not statistically significant. Almost half the cases of hepatitis affected several members of the family. The results suggest that viral hepatitis A in adult age has a more severe course than in children. The authors recommend in cases with an elevated transaminase serum activity more frequent check-up examinations to avoid missing of a relapse, and to examine repeatedly IgM anti-HAV as in protracted forms of hepatitis IgM anti HAV may persist even when the transaminase activity is normal. PMID- 7483335 TI - [New aspects of pharmacologic and general prophylactic care of the diabetic foot]. AB - Ischaemia, neuropathies and infections are predisposing factors for the development of ulceration of the diabetic foot. Diabetics have evidently a disposition for affections of the peripheral circulation and impaired regulation of the microcirculation as a result of autonomic neuropathy. The lower the driving pressure (in critical ischaemia), the more important are rheological factors and drugs which can influence them. These preparations include e.g. Trental (pentoxiphilline), Prostavasin (prostaglandin E1), Vessel due F (sulodexide). In advanced stages of ischaemic extremities oedema is a very adverse factor. Non-cardiac oedema can be very effectively handled by manual lymphatic drainage combined with intermittent one-segment pneumatic compression which was successfully used by the authors in ulcerations of the diabetic foot. One of the main general protective measures is adequate care of the foot and protective footwear for diabetics. After 3.5 years' use of protective footwear the authors recorded, consistent with data in the literature, a 50% reduction of relapses of ulcerations (and amputations). By examination on an EMED II apparatus abnormally high local pressures on the sole of risk patients can be detected and at the some time the protective effect of materials used for protective insoles can be tested. Active pharmacological and generally protective care of diabetic foot leads to a reduced number of amputations, in particular supracondylar ones by 50 or more per cent. PMID- 7483336 TI - [A vasoactive-intestinal-polypeptide producing tumor (VIPoma) as an uncommon cause of life-threatening hypokalemia]. AB - The authors present a case of a 40-year-old female patient examined repeatedly in the course of 11 years and hospitalized on account of pain in the hypogastrium, subfebrile temperatures, watery diarrhoea, hypokaliaemic alkalosis, weakness, fatigue and loss of body weight. As to laboratory examinations hypokaliaemia, hyponatraemia, metabolic alkalosis, irregularly elevated CRP values and minor leucocytosis predominated. A weight loss of cca 8 kg along with a severe mineral deficiency and clinical symptomatology called for parenteral nutrition with a mean daily substitution of 240 mmol K and 200 mmol Na. Due to the clinical condition and non-specific results of graphic and histological examinations the possibility of a VIPoma was considered. This diagnosis was confirmed by laboratory examinations and clinically--after the onset of corticoid treatment marked improvement of the general condition occurred. Finally the authors discuss diagnostic and in particular therapeutic possibilities in this disease. PMID- 7483337 TI - [Functional activity of a metastatic differentiated carcinoma of the thyroid gland]. AB - In a patient with differentiated thyroid carcinoma of the follicular type after removal of the thyroid gland and nodular secondaries on the neck a functional secondary was detected in the liver by combined whole-body scintigraphy following administration of 131-I and scintigraphy of the liver using 99mTc-colloid, supplemented by scintigraphy of the skeleton following administration of 99mTc MDP. At first the high thyroglobulin serum level was after repeated radioiodine treatment falsely negative. The author presents a review of cases reported in the literature who suffered from this rare type of secondaries. PMID- 7483338 TI - [Arrhythmogenic right ventricle]. AB - A 26-year-old patient with an irrelevant personal and family-history was treated on account of attacks of relapsing ventricular tachycardia and cardiac failure, mostly dextrolateral. After ruling out other possible causes of the disease--such as embolism of the lungs, ischaemic heart disease, out other possible causes of the disease--such as embolism of the lungs, ischaemic heart disease, Ebstein's anomaly, the diagnosis of an arrhythmogenic right ventricle was established. This disease is suggested by negative T waves in the thoracic leads V1-V3 during sinus rhythm and by the shape of the QRS complex which was the type found in block of the Tawara branch with an axis of + 100 degrees during attacks of ventricular tachycardia. Moreover, ventricular tachycardia of the same type was produced during electrocardiographic examination. On angiographic examination ARVD is suggested by marked trabeculization and impaired kinetics of the outflow tract of the right ventricle. In the prevention of relapses of ventricular tachycardia beta-blockers and Cordarone were successful when used simultaneously with cardiac stimulation type AAI applied on account of a confirmed sinoatrial block. PMID- 7483339 TI - [Determination of serum lactic acid and the acid-base equilibrium in carbon monoxide poisoning]. AB - The authors present a description of severe accidental carbon monoxide poisoning. They stress the significance of the examination of lactic acid level at this poisoning. In case described the level of carbonyl haemoglobin could not be determined because of device trouble. Patient AA (16-year-old) was admitted in relatively very good condition, without clinical signs of severer affection. Considering the facts that gas-poisoning (over 24 hrs) was found through anamnesis, and simultaneous admission of patient's greatgrandmother in critical condition, the patient AA was treated also with oxygen of high concentration, monitored, kept in absolute rest. High lactate level (18.9 mmol/l) and the evidence of metabolic acidosis (pH 7.16, pO2 8.65, pCO2 2.06, sBE -22.1) made to suspect of severe poisoning. Despite intensive oxygenotherapy it came to severe left-side cardial failure associated with pulmonal edema, anuria, shock after 4 hours, managed successfully within 4 hours. Lactate level correlated very well with condition severity. Septum hypokinesis was demonstrated cardiografically and it disappeared within 6 days. However, symetric inversion of T wave in many leads (I, II, aVF, V 2-6) persisted for 24 days. The authors assume that routine examination of acid base balance and lactate level in addition to the determination of carbonyl haemoglobin level is very important for the consideration of the severity of carbon monoxide poisoning. Lactataemia determination is probably even more significant for the assumption of oxygen deficit and therefore for the total prognosis of the patient. PMID- 7483340 TI - [Proton pump blockers and their significance in gastroenterology]. AB - Extremely effective drugs used in the therapy of diseases etiologically associated with excessive HCl secretion by the gastric mucosa include proton pump blockers. Their obvious advantage, as compared with commonly used drugs, is that they cause a marked drop of HCl production by the gastric mucosa regardless of the type of stimulus, i.e. whether parietal cell receptors are influenced. Their therapeutic action is extremely effective, while side-effects are minimal. The publication gives the basic indications of administration of proton pump blockers, compares their therapeutic efficiency with H2 blockers and draws attention to possible pitfalls of therapy. PMID- 7483341 TI - [Regulation of HDL lipoprotein plasma levels in relation to atherogenesis]. AB - There is no doubt nowadays on the importance of HDL lipoproteins in atherogenesis. In the submitted review the author presents information on the regulation of HDL lipoprotein plasma levels and their sub-classes. First he deals with metabolic relations, in particular synthesis and catabolism of HDL lipoproteins. He mentions then factors which influence plasma levels of HDL lipoproteins and their sub-classes--external and internal factors. As to internal factors the author discusses genetic predisposition, sex, age, obesity, adipose tissue distribution; as to external factors the effect of diet, exercise, alcohol, smoking and drugs. PMID- 7483342 TI - [Angiotensin converting enzyme inhibitors in the treatment of arterial hypertension]. AB - The author presents a brief account on mechanisms of action of ACEI in arterial hypertension. He mentions different representatives of ACEI, incl. new pharmacological preparations. He mentions the basic strategy of treatment, incl. clinical indications of the use of ACEI in hypertension and suitable and less suitable combinations of drugs. In the conclusion he mentions the most frequent side-effects of ACEI, absolute and relative contraindications of ACEI. PMID- 7483343 TI - [The molecular basis and in vitro determination of multiple resistance to cytostatic agents]. AB - Contemporary therapeutic procedures used in the treatment of malignant diseases are based on the no-selective action of cytotoxic or cytostatic substances. The sensitivity of tumour cells to cytostatics can be, however, reduced by the action of several mechanisms among which expression of the membrane glycoprotein P-170 is the most important. The latter actively transfers the cytotoxic substance from the cell and is responsible for multiple drug resistance-MDR). Glycoprotein P-170 can be assessed on leukaemic cells immunochemically by flow cytometry. A very perspective procedure is assessment of the in vitro sensitivity of leukaemic cells to clinically used cytostatics. PMID- 7483344 TI - [Sigismundus Albicus, the first rheumatologist at Charles University in Prague (1358-1427)]. PMID- 7483345 TI - [Changes in internal medicine over the past nine decades]. PMID- 7483346 TI - [Importance of programmed ventricular stimulation in the evaluation of proarrhythmic effects of orally administered anti-arrhythmia agents]. AB - The objective of the submitted work was to assess, using programmed stimulation of the cardiac ventricles during antiarrhythmic treatment (amiodarone and propafenon administered orally), the presence of indicators suggesting imminent ventricular ectopic proarrhythmia. The authors included in the group, after elimination of subjects with ECG (Holter monitoring) signs of proarrhythmias 50 patients (43 men, 7 women, 23-78 years, mean age 54.0 +/- 14.2 years), where they used initial programmed stimulation of the ventricles, not sooner than three months after acute cardiac disease. On account of electric instability of the ventricles the authors administered to 39 patients amiodarone by the oral route (200 to 400 mg/day) and to 11 subjects propafenon also by the oral route (900 mg/day). A subsequent programmed stimulation of the ventricles was repeated after amiodarone treatment after an interval of 4-144 weeks and after propafenon treatment after 1-48 weeks. They included among indicators of proarrhythmic action of antiarrhythmics (as compared with the initial examination): induction of diagnostic tachycardia at a lower grade of stimulation or with a smaller number of extrastimuli, the change from non-persisting unimorphous ventricular tachycardia into permanent unimorphous ventricular tachycardia, an increase of the frequency of tachycardia, marked deterioration of the patient's symptoms during tachycardia and the necessity to eliminate tachycardia by an electric discharge (during the initial examination an electric discharge was not used). The authors described in the group on indicator of proarrhythmic effect conditioned by programmed stimulation of the ventricles in 24 patients (48%). In one subject they found simultaneously two indicators in 12 instances (24%) and three indicators in 5 instances (10%).(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7483347 TI - [Programmed esophageal stimulation of the atrium and the long-term effects of anti-arrhythmia therapy of atrial fibrillation and flutter]. AB - The authors investigated the long-term effect of antiarrhythmic treatment on the maintenance of the sinus rhythm in a group of 32 patients with paroxysmal atrial fibrillation and flutter where the antiarrhythmic drug was selected because of negative results of tests by programmed atrial stimulation, using the method of oesophageal stimulation. After a one-year follow up 56% of the patients had permanently a sinus rhythm without paroxysms of supraventricular arrhythmia. Comparison with results of work evaluating the effectiveness of different antiarrhythmic drugs as regards maintenance of the sinus rhythm without testing suggests that selection of an antiarrhythmic drug by programmed stimulation of the atria by oesophageal stimulation has no predictive value for the effect of antiarrhythmic treatment. PMID- 7483348 TI - [Safety of maximal ergometric loading in a population of 37,250 individuals at risk (1969-1994)]. AB - No fatal complications occurred in 37,250 exercise tests (98.7% symptom limited) including 683 predischarge maximal tests in patients soon after myocardial infarction. Acute myocardial infarction was observed in 4 patients, ventricular fibrillation in 1 patient was successfully resolved by defibrillation. Extension of infarct area was recorded in 1 patient soon after myocardial infarction. Complication rates decreased from 1:3,400 in years 1969-1984 to 11,700 in years 1985-1994. Maximal exercise testing soon after myocardial infarction appears to be safe too. It cannot be replaced by submaximal testing. It appears to define high-risk patients suitable for revascularization procedures. PMID- 7483349 TI - [Serum interleukin-6 in multiple myeloma: I. Relation to selected laboratory indicators of disease]. AB - In a group of 111 patients with multiple myeloma (MM) comprising a group of 34 patients examined when the diagnosis was established and a group of 77 patients evaluated in different stages of the disease, the author examined the relationship between the interleukin-6 serum level (IL-6), assessed by the method of enzyme immunoanalysis and selected laboratory indicators of the disease. Elevated IL-6 values were recorded in 38% of the patients. In neither of the groups significant relations were found between IL-6 and calcium, urea, creatinine levels, the amount and type of monoclonal immunoglobulin, lacticode dehydrogenase, beta 2-microglobulin, ferritin, IL-2 and its soluble receptor in serum and the incidence of myeloma plasmocytes in bone marrow. In the second (but not in the first) group a significant relationship was recorded between IL-6 levels and the red cell sedimentation rate, the Hb value, the CRP level and serum albumin and the value of thymidinekinase in serum of patients with a value beyond the normal range. From the investigation ensues that examination of IL-6 serum levels in MM contributes so far mainly to improvement of the diagnosis and expedient classification of this disease in clinical practice. PMID- 7483351 TI - [Betaxolol hydrochloride in the treatment of mild and medium hypertension--long term effects]. AB - All available beta-blockers are effective in the treatment of high blood pressure. Differences in the pharmacokinetics are important as they can have an impact on their clinical use. Betaxolol, a beta-1-selective blocker has a long biological half-life which can control the blood pressure and pulse rate for 24 hours. The authors investigated 20 patients with mild and medium severe hypertension by out-patient monitoring of the blood pressure. Treatment was focused on influencing the morning rise of blood pressure and pulse rate. The results confirm the effectiveness of betaxolol (Lokren) in the administered dose of 10-20 mg once per day. A favourable response was obtained in 75% when checked by a sphygmomanometer. The morning readings of blood pressure at 5, 6 and 7 o'clock following treatment did not exceed 90 mmHg diastolic pressure. It was also found that there is a statistically significant difference in the effect exerted on pressure and pulse rate in daytime, while the difference at nighttime is not significant. This mild reduction on blood pressure and pulse rate during the night is an advantage in particular in elderly patients. Lokren treatment did not have any serious side-effects it did not affect laboratory values or ECG with the exception of the pulse rate. PMID- 7483350 TI - [Serum interleukin-6 in multiple myeloma. II. Relation to activity and stage of disease]. AB - In a group of 111 subjects with multiple myeloma (MM) comprising a group of 34 patients examined when the diagnosis was established and a group of 77 patients examined in different stages of development of MM the authors evaluated the relationship between interleukin-6 (IL-6) serum levels and the clinical activity and the stage of the disease. In both groups a significant relationship was found between IL-6 and the clinical activity of MM; "stable" and "active" stages of the disease differed by the frequency of elevated values and the level of IL-6. In both groups the authors recorded rising levels and a rising rate of subjects with elevated IL-6 levels with advancing stages of the disease. When staging systems were used according to Durie-Salmon, the British Medical Research Council and according to Bataille the highest IL-6 values were recorded in the third stage of the disease, these values being significantly higher than in stages 1 and 2. In the group assembled at the time of assessment of the diagnosis of MM the described differences did not reach (with the exception of the evaluation according to Bataille) statistical significance. The classification of patients in stages 1-3 into sub-groups with regard to the activity of the disease ("stable" and "active") was associated with significantly different IL-6 levels. The investigation revealed that examination of IL-6 levels contributes at present rather to the understanding of the pathogenesis and biology of MM than to practical evaluation of the severity, activity and stage of the disease. PMID- 7483352 TI - [Non-steroidal antirheumatic ointments in the treatment of primary periarticular and intramuscular fibrositis]. AB - The author presents his experience assembled in a short-term (two-week) therapeutic trial with Mobilisin ointment, which contains flufenamic acid, in 50 patients with primary localized periarticular and muscular fibrositis. The preparation was used locally three times per day-2-3 cm of ointment pressed from a tube. The patients were given a form in which they recorded every night the effect classified in four grades from the best to zero effect. In 49 the tolerance of the ointment was very good. In 45 patients a therapeutic effect was achieved (90%), comprising 21 (42%) where the effect was excellent, in 24 it was good or satisfactory (48%) and only in 5 patients no effect was recorded or the patents' statement was vague. In the first group of 21 patients 16 were able to abandon analgetic and anti-inflammatory treatment with non-steroid antirheumatics. Patients with the muscular form of fibrositis practically agreed in favour of the ointment. Gels according to these patients tissues. The authors remind that according to their experience gels, incl. Mobilisin, are suited for periarticular forms of fibrositis. It is also useful to rub the ointment in micro massage to achieve a greater initial hyperaemia and better resorption. Mobilisin ointment may prove useful also in other disciplines, in particular orthopaedics, surgery, traumatology, neurology, rehabilitation and sports medicine. PMID- 7483353 TI - [Cardiac impairment detected by echocardiography in patients with ankylosing spondylitis]. AB - The authors subjected to complete echocardiographic examination 40 patients with ankylosing spondylitis (AS) in order to assess pathological changes of the heart (changes of the structure and function of valves, myocardial function of the left ventricle and the pericardium) and compare their incidence with clinical characteristics of the patients (age, duration, stage, form and activity of the disease) as well as with the presence of ECG changes. Based on the results and their statistical analysis they revealed that structural changes of the aorta and its valves associated with dilatation of the root of the aorta, its mild insufficiency and the possible finding of a typical bump on its posterior wall is found in half the patients with AS, being more frequent when the disease develops early, when it persists for a long time or is in an advanced stage. A similar relationship is found also in the presence of mild diastolic dysfunction of the left ventricle. The authors did not observe a more frequent incidence of other pathological changes of the heart in patients with AS. None of the detected changes were clinically severe and did not call for aimed treatment or surgery. In the conclusion the authors compare the assessed facts with data in the literature and confront data in the literature. PMID- 7483354 TI - [Lipoprotein (a): a genetic risk factor with atherogenic and thrombogenic properties]. AB - Lipoprotein (a) is a relatively new independent risk factor of early atherosclerosis with atherogenic and thrombogenic properties. From the structural aspect it resembles LDL-lipoprotein and differs from the latter by the presence of another glycoprotein-apolipoprotein (a). Due to the great similarity of apolipoprotein (a) and plasminogen, lipoprotein (a) is bound to plasminogen receptors on the fibrin surface (fibrinogen) and thus prevents the cumulation and activation of local fibrinolysis. Its levels are under strict genetic control and are very little influenced by external factors and available hypolipidaemic treatment. There is a great interindividual variability of lipoprotein(a) concentrations which is due above all to the structural variability of apolipoprotein (a). At present at least 34 isoforms of apolipoprotein(a) were described which differ as to the size of the molecule. This great structural variability has an impact not only on the function and pathogenicity of lipoprotein (a) but also on methods of its assessment. High lipoprotein(a) concentrations are found in subjects with early clinical manifestations of atherosclerosis, in nephrotic syndrome, in chronic renal insufficiency, in haemodialyzed patients and other diseases. They rise in women after the menopause and are favourably influenced by hormonal substitution therapy. There is a number of immunochemical methods used for its estimation which are very well reproducible within the same laboratory. The high interlaboratory coefficients of variation indicate, however, that unification of lipoprotein(a) analyses is urgent. PMID- 7483355 TI - [Factors with insulin-like effects: IGF-I and GLP-1]. AB - In the submitted review the author discusses two substances secreted into the circulation which can similarly as insulin lower the blood sugar level. These substances are IGF-I (insulin-like growth factor I) and GLP (glucagon-like peptide). While in case of the former it is not certain whether it participates in the glucose homeostasis, this is beyond doubt in the latter. IGF-I prepared by the recombinant technique can be used therapeutically in cases of insulin resistance caused by a receptor or postreceptor disorder, because it may act via its own receptor. Side-effects after larger doses are a problem. GLP-1, the use of which would be useful in type 2 diabetics as it stimulates insulin secretion, is not used so far in therapy because hitherto prepared preparations have a very short period of a effectiveness. PMID- 7483356 TI - [Electrocardiographic Holter monitoring in patients with ventricular pre excitation: possibilities of its uses and pitfalls]. AB - Holter monitoring (HM) of the electrocardiogram (ECG) is used currently for the management of patients with ventricular preexcitation (PE). It makes it possible to detect inconstant PE which is considered a marker of a favourable clinical development. HM of ECG is not suitable for the detection of paroxysmal supraventricular tachycardias (PSVT) in patients with PE, as their presence is detected only exceptionally during HM. It can be used for the detection of other disorders of the cardiac rhythm than PSVT, in particular in supraventricular and ventricular extrasystoles (SVES and VES). Frequent SVES and VES detected during HM can as potential triggers of PSVT indicate a higher risk of these arrhythmias. HM is used for the objectivization of relatively frequent, sometimes quite polymorphous subjective complaints of patients with PE. Subjective data of these patients are very unreliable and from these data it is not possible to draw conclusions on the presence or absence of disorders of the cardiac rhythm. HM of ECG is not a suitable method for the evaluation of the effectiveness of medicamentous antiarrhythmic treatment of PSVT because of their considerable spontaneous variability. It may be, however, used partly for the evaluation of success of radiofrequency ablation treatment. Evaluation of long-term ambulatory ECG Holter tracings in patients with PE is more complicated and more time consuming than in subjects without PE. In any case it is essential that the record should be evaluated by an experienced physician with sufficient knowledge in arrhythmology. PMID- 7483357 TI - [Endogenous digitalis-like substance. I. Basic characteristics]. AB - Endogenous digitalis-like substances (EDLS) present endogenous products which structurally belong to cardiac glycosides (CG). According contemporary views these substances potentially present a newly discovered hormonal group produced by adrenal cortex. EDLS, similarly exogenous CG, bind to specific CG- receptor of Na(+)-K(+)-ATPase and inhibit activity of this enzyme. At the cellular level, EDLS lead to increase in intracellular calcium storing and such may play an important role in modulation of calcium-dependent processes and their response to characteristic stimuli (e.g. contractility of vascular smooth muscle cells, hormonal secretion etc.). First part is a review of present views to character and cellular effects of EDLS. PMID- 7483358 TI - [Lyme carditis]. AB - The authors present in the submitted paper the basic characteristics of Lyme carditis as one of the forms of Lyme borreliosis. They mention briefly the clinical symptoms, diagnostic possibilities and causal and symptomatic therapeutic procedures. In the second part they present conclusions which they obtained by comparison of data in the literature and their own experience with dispensary care of 50 patients during several years. In the conclusion they emphasize the great interindividual variability of the clinical picture not only in the acute stage of the disease but also for years after treatment. PMID- 7483359 TI - [On the integrity of internal medicine (particularly in type II hospitals)]. PMID- 7483360 TI - [The organizational characteristics of the medical support for the troops in the first operations at the start of a war (based on the experience of exercises)]. AB - The article summarized the experience of an explorative tactical-special exercise on medical support of Mobile Forces in first military operations of the initial period of war which has studied the following questions: the organic structure and organization of work of the medical service of a motorized infantry brigade in defense; joint direction of organic hospital facilities and field traumatological hospital. The author also studies the deployment peculiarities of a military multipurpose hospital on the basis of a garrison hospital. The experience of these exercises has proved the necessity to update the organic structure and principles of the employment of medical assets in accordance with the requirements of new military doctrine, as well as realize the technical re equipment of medical service. PMID- 7483361 TI - [The optimization of the management of a general hospital]. PMID- 7483362 TI - [The dispensary care of servicemen utilizing the territorial principle in providing dental care]. PMID- 7483363 TI - [Combat psychic trauma and the provision of psychiatric care in light casualties at medical evacuation stages]. PMID- 7483365 TI - [Medical support for the Soviet Army in the operations of World War II]. PMID- 7483364 TI - [The diagnosis and treatment of light casualties with involvement of the organs of the urogenital system]. PMID- 7483367 TI - [Gastric evacuatory function following a Roux resection]. PMID- 7483366 TI - [Roux gastric resection in peptic gastroduodenal hemorrhages]. PMID- 7483368 TI - [The use of the atlanten nutrient mixture in artificial feeding]. PMID- 7483369 TI - [The treatment of middle-aged and elderly patients at a military polyclinic]. PMID- 7483370 TI - [An analysis of the epidemic health status of the troops and the basic trends in improving medical prophylaxis]. PMID- 7483371 TI - [A hygienic analysis of the irradiation doses of those cleaning up the aftermath of the accident at the Chernobyl Atomic Electric Power Station (based on data from the All-Army Registry)]. AB - In the result of the analysis of irradiation doses of 36,895 servicemen who have participated in liquidation of the Chernobyl APC disaster consequences it was found out that the average radiation dose was 12.1 cSv in 1986, 8.4 cSv in 1987, and in 1988-90 it didn't exceed 5 cSv. The irradiation received by these servicemen has depended on the time of their arrival to the zone of the disaster, the maximum permissible doses of radiation established at that period, the character of their activities and the place of their work, utilization of protection means. A conclusion was made that irradiation dose itself could be considered only as a general guide reference, because it didn't take into consideration internal irradiation and beta-irradiation of skin. That's why it is necessary to continue the further monitoring over the health status of liquidators independently of the doses of irradiation which were fixed in their histories. PMID- 7483373 TI - [The diagnosis of premorbid psychosomatic states in ship-based specialists]. AB - The formation of premorbid state was traced in 89 seamen from the whole crew number of 399 by the end of cruise. In 41 seamen abdominal microsystems have appeared for the first time in their life 2-3 weeks after the development of preneurotic state in the course of cruise. The following terminology is proposed: forming psychosomatic abdominal syndrome in combination with preneurotic state with microsymtomocomplex, which didn't obtain the level of functional disorders of stomach, intestinal or biliary tract, and formed psychosomatic abdominal syndrome to denominate the combination of preneurotic or neurotic state with some functional disorders of stomach, intestinal or biliary tract. It is recommended to apply psychotherapy during treatment, primary and secondary prophylaxis in seamen. PMID- 7483372 TI - [The effect of training flights on the level of statokinetic resistance in the aviation cadets of foreign armies]. AB - The article studies the influence of training flight upon the statokinetic resistance of cadets from foreign countries. It was found out that in the process of flight training the level of statokinetic resistance was growing, and simultaneously the vegetative, sensor and somatic manifestations were decreasing, and the period of stable equilibration in N.N. Bondarevskii test was increasing. The most pronounced changes in statokinetic resistance was marked in the cadets from Mozambique, Mongolia and Angola; the smallest--in cadets from Vietnam and Laos. In order to optimize the flight training of cadets and safety of training flights it is recommended to apply the system of complex medical monitoring, which will realize the control over the psychophysiological state and statokinetic resistance during the whole period of training. PMID- 7483374 TI - [The history of the organization of medical care in Soviet prisoner-of-war camps]. PMID- 7483375 TI - [An analysis of the acupuncture treatment results in bronchial asthma patients]. AB - Treatment effects reached in 94 patients with bronchial asthma demonstrate that neurogenic, humoral and bioenergetic responses to acupuncture proceed according to adaptation laws and result in reduction of bronchial hyperreactivity. Eosinophilic inflammation in the bronchi diminishes acupuncture efficacy. PMID- 7483376 TI - [An evaluation of the effect of magnetic-laser therapy on external respiratory function in complicated forms of acute pneumonia in children]. AB - The authors studied the effects of magnetic laser therapy (MLT) on external respiration, blood transport of gas and clinical course of acute destructive pneumonia in children aged 1-12 years. It was established that energy of infrared laser combined with magnetic field reduces the severity of acute respiratory insufficiency, treatment course, prevents destructive complications in children with infiltrative acute destructive pneumonia. PMID- 7483377 TI - [Experience in using hydrotherapy in children with acute lymphoblastic leukemia at a sanatorium]. AB - 34 children aged 10-15 years in long-term remission of acute lymphoblastic leukemia were on combined rehabilitation for concomitant diseases. Adjuvant balneotherapy promoted improvement in the heart rate and decreased asymmetry of circulation. No side effects were registered. PMID- 7483378 TI - [Physiotherapeutic methods in the postoperative period in acute pancreatitis patients]. AB - 60 patients surgically treated for acute pancreatitis received postoperative physiotherapy which combined transcutaneous electroneurostimulation with exposure to low-energy laser. The results support the effectiveness of combined physiotherapeutic methods of general and reflex action in severe forms of acute pancreatitis because such physiotherapy early after severe forms of acute pancreatitis because such physiotherapy early after severe pancreatitis surgery promotes the decline of pathological symptoms. PMID- 7483379 TI - [An experimental validation of hyaluronidase electrophoresis for the prevention of postoperative adhesions]. AB - The experiments on 50 animals were made to evaluate efficacy of electrophoresis of hyaluronidase (from actinohyal solution) against adhesions. Model abdominal adhesions in rats were exposed to hyaluronidase coming from anode and cathode for 20 min. There were altogether 10 daily procedures. The control animals received the solvent from the same poles or were not exposed to electrophoresis. The results demonstrate that hyaluronidase introduction from anode and cathode significantly decreases the occurrence and density of postoperative adhesions compared to controls. PMID- 7483380 TI - [The effect of electrical and biomechanical [correction of biochemical] stimulation on metabolic processes in the skeletal musculature (experimental research)]. PMID- 7483381 TI - [The rehabilitation of disabled children--a government-wide problem]. PMID- 7483382 TI - [The intensification of the activities of sanatorium-type institutions]. PMID- 7483383 TI - [The morphological aspects of the anti-ischemic effect of a low-intensity helium neon laser on the coronary vessels (experimental research)]. PMID- 7483384 TI - [The reactions of the cardiovascular system in children suffering from inflammatory lung diseases to cold exposures]. PMID- 7483385 TI - [The role of chronotherapy in optimizing the treatment of patients living under ecologically adverse conditions]. AB - The problem of chronobiological optimisation deserves special attention of medical specialists as promising in chronotherapy and chronopharmacology. Its solution requires much effort and adequate methodological approaches. PMID- 7483386 TI - [Magnetotherapy in the treatment of duodenogastric reflux in patients with gastric and duodenal peptic ulcers]. PMID- 7483387 TI - [A box for performing air-carbon dioxide-radon baths]. PMID- 7483388 TI - [The dynamics of the physical work capacity of women during pregnancy]. PMID- 7483389 TI - [A new deposit of radon waters]. PMID- 7483391 TI - [The Central Institute of Health Resort Medicine on the eve of and during World War II]. PMID- 7483392 TI - [The theoretical bases of rehabilitation in exotoxicosis]. PMID- 7483393 TI - [The mechanism of action of a magnetic field on the living organism]. PMID- 7483390 TI - [The organization of sanatorium-health resort treatment in hospital bases at the rear in World War II]. PMID- 7483394 TI - [On the centenary of the birth of A. N. Obrosov]. PMID- 7483395 TI - [The problems and prospects of a climatotherapy health resort under current conditions]. AB - The experience of climatic health resorts at the Crimea sea shore has been summarized. The potential of climatic treatment can be fully realized only with the participation of therapeutic, rehabilitation and recreation institutions. PMID- 7483396 TI - [The meteoropathic reactions of patients with cardiovascular diseases and the methods for their prevention with contrasting temperature exposures]. AB - High sensitivity of patients with cardiovascular diseases to weather conditions was successfully managed by contrast temperature training which secured not only resistance to temperature fluctuations but also to a complex of weather factors. This, in the long run, secured a decline in the number of coronary heart disease and hypertension exacerbations and higher quality of life. PMID- 7483399 TI - [The current status of the boron neutron capture therapy of tumors]. PMID- 7483398 TI - [The technical and technological support of modern methods for radiation diagnosis and therapy in tumors]. PMID- 7483397 TI - [The radiosurgical organ-preserving treatment of breast cancer]. PMID- 7483401 TI - [Modern means for planning radiation therapy]. PMID- 7483400 TI - [Medical linear accelerators. The features of their use and technical characteristics]. PMID- 7483402 TI - [The organizational problems of cancer care today]. PMID- 7483403 TI - [Medical physics. The problems of training personnel]. PMID- 7483404 TI - [The interstitial radiation therapy of cancer of the tongue and mucosa of the floor of the mouth]. PMID- 7483405 TI - [The use of x-ray computed tomography and a simulator in the planning of radiation therapy]. PMID- 7483406 TI - [The current potentials and outlook for drug therapy in oncology]. PMID- 7483407 TI - [The individualization of the drug therapy of ovarian cancer]. PMID- 7483409 TI - [The neoadjuvant therapy of malignant tumors]. PMID- 7483408 TI - [The adjuvant hormonal therapy of breast cancer]. PMID- 7483410 TI - [The maintenance treatment of patients with stomach cancer who have undergone a radical surgical intervention]. PMID- 7483412 TI - [Neoadjuvant drug therapy as a predictor of the efficacy of castration]. PMID- 7483411 TI - [The determination of individual sensitivity to cytostatics in breast cancer]. PMID- 7483413 TI - [New antitumor preparations]. PMID- 7483415 TI - [Physical action factors in the treatment of the initial forms of cervical cancer]. PMID- 7483414 TI - [The photodynamic therapy of malignant tumors in basic sites with the preparations photohem and photosens (the results of 3 years of observations)]. PMID- 7483416 TI - [First experience in using a whole-body magnetic field in treating cancer patients]. PMID- 7483417 TI - [Nondamaging laser therapy in the combined treatment of tumors]. PMID- 7483418 TI - [The potentials of a complex of physical factors in the surgery of tumors of the trachea, lungs and mediastinum]. PMID- 7483419 TI - [The Urals Center of Neutron Therapy]. PMID- 7483420 TI - [Cancer care for the population of Arkhangel'sk Province and the ways for its improvement]. PMID- 7483421 TI - [The role and tasks of a provincial (regional) cancer center in improving cancer service]. PMID- 7483422 TI - [The role of social organizations in improving the fight against cancer]. PMID- 7483424 TI - [New approaches in the organization of the cancer service of Bashkortostan]. PMID- 7483423 TI - [The distinctive characteristics of malignant neoplasms in the population of the Altai Territory subjected to long-term radiation exposure as a consequence of the multiyear nuclear explosives tests at the Semipalatinsk proving ground]. PMID- 7483425 TI - [The organization of palliative care for cancer patients and the outlook for its improvement]. PMID- 7483426 TI - [Malignant thyroid neoplasms in the territories of the Russian Federation exposed as a consequence of the accident at the Chernobyl Atomic Electric Power Station]. PMID- 7483427 TI - [The development of a state anticancer program]. PMID- 7483428 TI - [The potentials and prospects of modern teaching and testing methods in training students and physicians in oncology]. PMID- 7483429 TI - [The experimental chemotherapy of malignant tumors today]. PMID- 7483430 TI - [Immunological approaches to the use of cytokines in the combined treatment of malignant neoplasms]. PMID- 7483431 TI - [The pharmacological protection of the body from malignant tumor metastasis]. PMID- 7483432 TI - [The outlook for combining antitumor and activation therapies]. PMID- 7483433 TI - [The search for active antitumor agents among triphenylethylene derivatives]. PMID- 7483434 TI - [Nitroxyl radicals--modifiers of the toxic action of cytostatics]. PMID- 7483435 TI - [The biochemical mechanisms of drug resistance to N-alkyl-N-nitrosoureas and derivatives of cis-dichlorodiammineplatinum]. PMID- 7483437 TI - [Radiobiology and the progress of radiation oncology]. PMID- 7483436 TI - [Decreasing the toxicity of antitumor preparations--the way to enhancing treatment efficacy in malignant tumors]. PMID- 7483438 TI - [The radiobiological validation and clinical results of thermoradiotherapy in malignant tumors]. PMID- 7483439 TI - [The role of inhibition of the blood flow in a tumor undergoing antitumor therapy]. PMID- 7483440 TI - [Experience in using different radiobiological models for predicting the early and late radiation effects in the normal tissues of patients with primary and metastatic cancer of the vagina]. PMID- 7483441 TI - [The optimization of the procedure for the conservative therapy of tumors and the achievements of oncological biology]. PMID- 7483442 TI - [Intraoperative hypoxic radiotherapy (IOHRT)--its first trial in world practice]. PMID- 7483443 TI - [Technological approaches to the systemic radiation therapy of cancer patients]. PMID- 7483444 TI - [The current status of the problem of predicting tumor reaction to radiation treatment]. PMID- 7483445 TI - [A glance into the future (the outlook for pediatric oncology)]. PMID- 7483446 TI - [The postgraduate training of the pediatric oncologist]. PMID- 7483447 TI - [The current diagnostic and treatment aspects of primary malignant liver tumors in children]. PMID- 7483448 TI - [The treatment of nephroblastoma recurrences and metastases in children]. PMID- 7483449 TI - [Bone sarcomas in children]. PMID- 7483450 TI - [Diagnosis, surgical treatment and its results in thyroid cancer in children]. PMID- 7483451 TI - [Interferon and tactivin in the treatment program for non-Hodgkin's lymphomas in children during remission]. PMID- 7483452 TI - [Organ-preserving operations in the combined treatment of primary malignant bone tumors in children]. PMID- 7483453 TI - [Modern radiation therapy: the achievements and outlook]. PMID- 7483454 TI - [The radiation therapy of skin cancer (fractionation variants and methodology)]. PMID- 7483455 TI - [Interstitial radiation therapy in the combined treatment of malignant tumors]. PMID- 7483456 TI - [Radiomodifiers and the progress of radiation oncology]. PMID- 7483457 TI - [The potentials of intraoperative radiation therapy in improving the surgical results in tumors of the gastrointestinal tract]. PMID- 7483458 TI - [The radioprotective action of total gas hypoxia for tumors and normal tissues]. PMID- 7483459 TI - [The effects of whole-body electromagnetic hyperthermia on circulating CFU-GM and on plasma colony-stimulating activity in patients with lymphogranulomatosis]. PMID- 7483460 TI - [Local radiation damage: the characteristics of its pathogenesis, diagnosis and treatment]. PMID- 7483461 TI - [Effectiveness of non-starch polysaccharide crylan in experimental hyperlipidemia]. AB - In vitro experiments were shown that native ecologically pure non-starch polysaccharide crylan bind the bile acid. Crylan was found to decrease hyperlipidemia induced in rats by means of diet enriched with cholesterol and 6 methylthiouracil. This effect of crylan was more pronounced as compare with the action of native nonspecific enterosorbent polyfepan. PMID- 7483462 TI - [Specialized products for nutrition of children with mucoviscidosis]. AB - Mucoviscidosis is serious inherited disease with lesions of almost all secretory cells of mucosa and exocrine glands, following by disturbances of metabolism, digestion and nutritional status. Using of modern high effective pancreatic enzymes does not allow to reach a normalization of protein metabolism and digestion especially of fats. For success treatment of this disease the effective dietetic methods should be used including dietary intake of special food products which can supply enough energy and nutrients in small volume. The nutrient contents of a few foreign substitution of breast milk and food product for older children are given. The results of treatment of 86 children suffering from mucoviscidosis are presented. Milk proteins (casein, whey proteins), hydrolysates of milk protein and soy protein isolates are main resources of protein component in special food. Fat components are produced from middle-chain triglycerides (60 70%) and vegetable oils. Carbohydrates in special food are made from glucose polymers, glucose, fructose and sucrose. Positive results of clinical trials of foreign special food have been an example for creation of two home-produced food for treatment of children with mucoviscidosis: 1) adapted version "A" for children in age under 1 year, and 2) version "B" for older children. The products were created on basis of native milk protein with ratio casein to whey protein as 60:40. Middle-chain triglycerides compose 70-75% of fats. Carbohydrates are presented by dextrin-maltose (85%), glucose and sucrose. In present time these special products are under clinical trial in scientific medico-genetic center of Russia' Academy of Medical Science. PMID- 7483463 TI - [Organization of nutrition of children in preschool institutions]. PMID- 7483464 TI - [Study of the mutagenic activity of cadmium by the method of dominant lethal mutation]. AB - Calculation of dominant lethal mutations was executed in intact female mouse, mated with male on 1 and 6 week after single intragastric ingestion maximum tolerated dose CdCl2 (2 mg of Cd per kg of body mass). The increase of total, pre and postimplantation lethality was found within first 4 weeks of observation. This effect testifies the alteration of spermatogenesis cycle on late stages when reparation of DNA is absent. On early stages of spermatogenesis (5th and 6th weeks) the dominant lethal mutations were on the same level as in control group. PMID- 7483466 TI - [Mineral properties of roots and rhizomes of Sanguisorba oficinalis]. AB - The content of 25 mineral elements in roots and rhizomes of Sanguisorba oficinalis stored up in Kemerovskaja region were determined by the method of spectral emission and atomic-absorption spectroscopy. The data are important hygienic criteria for quality of raw materials. The content of Cu and Pb does not exceed the maximum permissible concentrations adopted in this country for these elements. The relatively high concentrations of some elements observed in separate samples reflect ecological conditions of growing place of this plant. PMID- 7483465 TI - [Fusarium toxins in the cereal crop in Russia (situation in 1993 and 1994)]. AB - The occurrence of deoxynivalenol (DON) and zearalenone in 495 samples of food wheat, barley and rye collected from different regions of Russia, and in 633 samples of fresh harvested wheat from Krasnodar region in 1993 and 1994 was surveyed. DON was detected in 97% of fresh harvested wheat samples in 1993, exceeding maximum tolerated level MTL) in 69% of samples analyzed. 23% of fresh harvested wheat samples were positive for DON and only in 5% of samples investigated DON concentration exceed MTL in 1994. Zearalenone was found in low concentrations in 3 of 154 wheat samples analyzed. Surveys of food wheat, barley and rye samples have shown that 23%, 7% and 3% of lots were positive for DON, respectively, in 1993. DON concentration exceed MTL in 14% of food wheat samples. The frequency of DON contamination of the 1994 food wheat samples was 6%. No mycotoxins were found in the 1994 food barley and rye samples analyzed. Almost all DON contaminated lots of food grain were collected from North Caucasus region. PMID- 7483467 TI - [Taurine in the composition of a new dry milk product Femilak-2 intended for improving nutrition of nursing mothers]. AB - The contents and production technology of new dry milk product "Femilak-2" were elaborated by Institute of Nutrition of Russia's Academy of Medical Science in collaboration with company "Nutritec". This product, enriched by essential amino acid taurine is intended for correction of dietary intake of lactating women (TU 92 11371 113 4 92). This special product is produced by Balta's (Ukraine) and Volkovyssk's (Byelarus) tinned dairy food factories. The method of introduction of taufon preparation in product and safe keeping of taurine during "Femilak-2"' production and storage are presented. PMID- 7483468 TI - [Problems of food product market]. PMID- 7483469 TI - [Project Russia's Children in action]. PMID- 7483470 TI - [Nutrition of premature infants: current concepts, problems and prospects (literature review)]. PMID- 7483471 TI - [Sweetener Sweetly--dietetic sweetness]. PMID- 7483472 TI - [Lipid transport in blood from the protein chemistry viewpoint]. AB - In agreement with the protein biochemistry principles apolipoprotein is considered to the only protein which: 1) forms protein-lipid complex (PLC) based on one lipid grade; 2) determines its functional significance; 3) causes the development of dyslipidemia at genetic disorder of quantitative and qualitative protein composition. The lipid transport in blood flow is based on high functional specifics of each of apoproteins; each apoprotein forms functionally separate PLC; each PLC has got one protein-vector; each protein-vector interacts with only one receptor. The basis of united cycle functioning in lipid transport is the difference of primary apoprotein structure. Cholesterol conducts an auxiliary function in triglyceride transport providing circulation in functional cycle. The lipid transport in blood flow is based first of all on protein chemistry principles and secondary--on lipidology principles. PMID- 7483474 TI - [The effect of fermented hydrolyzed whey enriched with lactates on the morphological structure of the internal organs in laboratory animals]. AB - Results of pathomorphological investigation of rat organs fed during 1 and 3 months the diet containing 8% of protein as milk whey protein partly hydrolysed by enzymes and enriched by lactates (SGOL-1) are presented. The conducted investigation have not revealed of pathological effect of a product SGOL-1 on morphological structures of investigated internal bodies and tissues of the laboratory animals. A increase of body mass of animals and contents of RNAS in cells of practically all bodies and tissues of the animals were also marked. PMID- 7483473 TI - [The effect of fatty compounds with different ratios of the polyunsaturated fatty acids of the omega-3 and omega-6 families on the expression of food-related anaphylaxis, on the liver cytochrome P-450 system and on 17-hydroxycorticosteroid metabolism in guinea pigs]. AB - The effect of fat composition with a different ratio of polyunsaturated fatty acids of families omega 3 and omega 6 on expression of experimental dietary anaphylaxis, liver monooxygenase system and metabolism of 17-oxycorticosteroides (17-OCS) in guinea-pig have been investigated. Fat components in experimental diets were presented by sunflower, ichthyene oils and mix of these oils in the ratio 1:1. A ratio of PUFA of a family omega 6 to omega 3 in diets were 64,0; 0,13 and 2,64. It was shown that a burden of dietary anaphylaxis were reduced significantly at the introduction in diets of sunflower oil.ulakcuu is authentically reduced. Simultaneously a marked increase of a ratios P-450B to P 450L and b5 to P-450 and reduction of a hexenal sleep duration were determined. The amount of excreted polar 17-OCS was also decreased in animals fed sunflower oil. Probable mechanisms of hypoallergenic action of dietary sunflower oil were conditioned by reorganization of liver cytochrome P-450 system and respective alterations of 17-OCS metabolism are discussed. PMID- 7483475 TI - [The characteristics of amino acid metabolism in premature infants on different types of feeding]. AB - Analysis of the free amino acids (AA) in a blood plasma of premature infants receiving kinds of baby food has allowed to reveal a feature of AA metabolism. A significant increasing of sum of the free AA and stable contents of aromatic AA have been found in blood of premature infants fed Prepiltti [correction of 'Prepilti'] that testified to adequacy of protein in Prepiltti [correction of 'Prepiltu'] to requirements of premature infants. Also it has been established different changes of concentration methionine and cystine in this group of the babies that confirms a hypothesis about immature of liver enzymes catabolizing a methionine. PMID- 7483476 TI - [Vitamins with antioxidant action and rheumatic diseases (a review)]. AB - The review of the modern literature about a role of essential dietary antioxidants and first of all of vitamins E, C, P, A and carotenoids in protection of cell components against of damage by excessive strengthening of free radical oxidation is submitted. Expediency of inclusion of antioxidant vitamins in complex therapy of rheumatic diseases is discussed taking into account the failure in an antioxidant system and excessive strengthening of free radical oxidation as important symptoms in development of rheumatic diseases. PMID- 7483477 TI - [An interview with Academician of the Russian Academy of Medical Sciences Mikhail Nikolaevich Volgarev]. PMID- 7483478 TI - [The effect of the modified preparation Komplivit on blood coagulation]. AB - Modified medicine "Komplivit" enriched by compounds of selenium, methionine and iodine showed more severe antioxidant effect than original "Komplivit". Ingestion of modified medicine to white rats during 12 days declined significantly hemocoagulation disturbances caused by thrombinemia and decreased frequency of lethal thrombosis. Authors suggest the usefulness of investigations of modified "Komplivit" in clinical cases of thrombinemia. PMID- 7483479 TI - [New developments in the prevention and treatment of atherosclerosis, ischemic heart disease, hyperlipidemia and other diseases]. PMID- 7483480 TI - [The psychotherapeutic aspects of treating food-related somatic diseases]. PMID- 7483482 TI - [Products for the diet therapy of young infants]. PMID- 7483481 TI - [Dietary plant phospholipids as stabilizers of oil solutions of beta-carotene]. AB - A study of stability of oil solutions beta-carotene to oxidizing damage is carried out. As stabilizers were used dietary plant phospholipids produced by Krasnodar's company ECOTECH in concentrations 5, 10, 20% in a combination with alpha-tocopherol. Oxidizing changes in samples were evaluated by determination of peroxide level ('peroxidation number') and contents diene conjugates. Effect of these antioxidants was studied in experiments with accelerated oxidation at 60 C in darkness during 15 days. Phospholipids entered in a sample acted as antioxidants. Reverse correlation was found between quantity added phospholipids an level of accumulated primary peroxidation products. It was shown that during experimental oxidation together with oxidizing damage of oil the loss of beta carotene was also occurred: without antioxidants on 68%, with alpha-tocopherol alone on 27%, with phospholipids and alpha-tocopherol on 34%. Phospholipids protect an oil from oxidizing damage not rendering of direct action on safety ( carotene being only as synergists in a combination with alpha-tocopherol. Phospholipids increase thus antioxidant potentiation of a product lying as consider in the basis of antioxidant action of beta-carotene. PMID- 7483483 TI - [Quality control is the common cause. Interview by G. Borisova]. PMID- 7483484 TI - [The vitamin allowance of the adult population of the Russian Federation and its changes in 1983-1993. 1. Vitamins C, E, A and carotene]. AB - The investigations carried out in 1983-1993 showed increasing of insufficiency of vitamin C and carotene in population of almost all investigated areas of Russian Federation. Insufficiency of vitamin C was found in 70-95%, pronounced deficit in 45-70% of adults. Deficiency of carotene was was detected in 30-90% of adults. Insufficiency of these nutrients is unfavourable long condition detected not only during spring but also in summer in different occupational groups of population. Vitamins A and E nutritional status of different groups of population are good enough in all seasons. PMID- 7483485 TI - Molecular genetics of the ABO histo-blood group system. AB - Molecular genetic study of the histo-blood group ABO system has elucidated the allelic basis of this genetic locus. Comparison of the nucleotide sequence has identified in the coding region differences which change amino acid sequences of the glycosyltransferases coded by these genes. Effects of the differences (mutations) on the specificity and activity of the glycosyltransferases have been examined. PMID- 7483486 TI - Sensitivity and specificity of three third-generation anti-hepatitis C virus ELISAs. AB - Three commercially available 3rd-generation anti-HCV ELISAs (Abbott, Murex and Ortho) were evaluated in various serum panels: (A) blood donor samples (n = 403) with 1st- or 2nd-generation anti-HCV ELISA (various manufacturers) positive test results; (B) non-A, non-B hepatitis patients (n = 212); (C) multitransfused patients (n = 253); (D) serial dilutions of HCV confirmed (RIBA and PCR) positive blood donors (n = 24), and (E) first-time blood donors (n = 1,055). All samples of panels A, B and C were tested in PCR and RIBA-2. In panels A, B and C, 398 samples were HCV PCR positive: all were detected by Abbott and Ortho, and 397 (99.7%) by Murex. The sample missed by the Murex ELISA showed an isolated anti C33c reactivity in RIBA-2. In panels A-C, 442 samples were RIBA-2 positive and all were detected by the 3 tests. With Probit analysis on results of panel D, no significant difference in sensitivity was observed between the 3 evaluated ELISAs. Specificities of Abbott, Murex and Ortho in 1,055 blood donors were 99.7, 99.3 and 99.9%, respectively (NS, chi 2). We conclude that the sensitivity and specificity of the 3 ELISAs are comparable although the C33c antigen in the Murex VK47 test should be improved. PMID- 7483487 TI - Hepatitis C virus genotypes among blood donors and their recipients in Iceland determined by the polymerase chain reaction. AB - Eight antibody-positive individuals were detected among 12,000 blood donations during the first year of screening blood donors for hepatitis C virus (HCV) antibodies in Iceland. All 8 were found to have a history of intravenous drug abuse. Six of these 8 individuals had previously donated blood to 27 patients who could be traced and examined for HCV infection. The great majority (23/27, 85%) of the recipients had demonstrable HCV antibodies. Furthermore, RNA analysis with the polymerase chain reaction showed that all patients with HCV antibodies had HCV RNA in their serum and in one hemodialysis patient without HCV antibodies viral RNA could be demonstrated. Genotyping of the HCV strains showed that the genotype of the donor was also identified in all but one of the infected recipients of his/her blood or blood products. This study, therefore, substantiates high infectivity of the HCV by blood or blood factor donation and shows that viremic HCV antibody-negative individuals exist. PMID- 7483488 TI - Performance evaluation of a particle agglutination test for antibody to human immunodeficiency virus 1: comparison with enzyme immunoassay. AB - A performance evaluation of a particle agglutination test (PAT), manufactured by Fujirebio Inc., Japan (Serodia-HIV), for antibody to human immunodeficiency virus type-1 (anti-HIV-1) was carried out and compared with a currently available enzyme immunoassay (EIA), manufactured by Genetic Systems Corp., USA, (HIV-1/HIV 2 EIA). Testing 2,878 Indian donor and patient samples, both tests showed 100% sensitivity and comparable specificity (PAT: 99.8%; EIA: 99.7% among donor samples). We conclude that PAT is a specific and sensitive test for anti-HIV-1; it is simple to perform and does not require sophisticated equipment. Hence it is suitable for mass screening of blood donors in a developing country like India, especially in rural areas where presently no HIV-testing facilities are available. PMID- 7483489 TI - In vitro cytokine treatment of B cell defects in HIV-infected hemophilia patients. AB - HIV-infected patients exhibit defects in B cell differentiation and in the IL-6 response of B cells, in association with autoantibody formation against T cells. These autoantibodies have been implicated as important factors in the development of immunodeficiency disease. As the restoration of defective B cell responses might prevent autoantibody formation and the resulting immunosuppression, we studied whether in vitro treatment with recombinant IL-2 (rIL-2), recombinant IL 4 (rIL-4) or recombinant IL-6 (rIL-6) might restore the response of B cells of HIV-infected patients. B cells of 6 HIV-negative hemophilia patients, 4 HIV positive patients at CDC stage II, III, 4 HIV-positive patients at CDC stage IV, and 6 healthy controls were tested in Staphylococcus aureus Cowan I (SAC-I) stimulated B cell cultures and Pokeweed mitogen (PWM)-stimulated allogeneic B and T cell cocultures. B cell differentiation was assessed in a reverse hemolytic plaque assay and by ELISA determination of IgM, IgG and IL-6 in culture supernatants. In vitro application of rIL-6 resulted in suppression of both elevated unstimulated and mitogen-stimulated B cell responses in a dose-dependent manner which was in part due to feedback inhibition. PWM- and SAC-I-stimulated IgG and IgM responses, respectively, could be restored after addition of 10 U/ml rIL-2 in HIV-negative patients, but not in HIV-positive patients. Addition of rIL 4 to cultures resulted in suppression of both unstimulated and mitogen-stimulated IL-6 secretion and B cell responses. Severely depressed B cell responses in CDC IV patients were not significantly affected by cytokine application. These results indicate that defective Ig responses in HIV-negative patients may be restored by rIL-2 treatment whereas HIV-induced B cell defects are not corrected by supply of T cell help or cytokines promoting B cell growth and differentiation. PMID- 7483490 TI - Cytokines in platelet concentrates prepared from pooled buffy coats. AB - Platelet concentrates (PC) prepared from pooled buffy coat (BC-PC) contain a variable number of leukocytes from different donors. We questioned whether storage of BC-PC can lead to a lymphocyte activation in the sense of a mixed lymphocyte reaction. BC-PC were prepared from four ABO-identical buffy coats and we undertook leukocyte analyses and measurement of different cytokines on days 1, 3 and 5 of PC storage (n = 72). Cytokine content was also determined in freshly prepared plasma (n = 48) and PC prepared by thrombapheresis (SD-PC) (n = 12). As control, we studied lymphoproliferation of pooled peripheral blood mononuclear cells from four individuals in 10 mixed lymphocyte cultures (MLCs) under optimal conditions. In the BC-PC, whole blood count and lymphocyte analysis showed a mean leukocyte contamination of 64 +/- 28 x 10(6) per unit with a proportion of lymphocytes of 66.7 +/- 13%. In the MLC, levels of interleukin-2 (IL-2) and interferon-gamma (IFN-gamma) were increased on day 3 and 5 of storage (p < 0.001). In a proportion of BC-PC, tumor necrosis factor-alpha (72.2%) and IL-2 (43.1%) were detectable immediately after preparation, whereas IFN-gamma (4.2%), interleukin-1 beta (4.2%) and interleukin-8 (11.1%) were only found in some BC PC. In all cases, initial values of cytokines did not increase during storage. Cytokine measurement in FFP and SD-PC showed similar results. The study demonstrates that cytokines are detectable in a variety of blood products immediately after preparation.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7483491 TI - Absence of serological surrogate markers for Trypanosoma-cruzi-infected blood donors. AB - Specific serological screening tests for Trypanosoma-cruzi-infected donors are not yet available and thus not routinely performed in North America. With the recent increase of Latin-American immigration to North America and Europe, there is a risk of transmission by blood products. In this study, we evaluated the possibility whether any of the serological screening tests currently recommended by the AABB could be used as a surrogate marker for this protozoarium. A group of 26,365 blood donors (male = 21,053 and female = 5,312) was analysed for the correlation of T. cruzi antibodies (TcAb) with other serological markers (HIV, HBsAg, ALT, HTLV-I/II, HCV, Anti-HBc, syphilis and unexpected hemoglobins other than A1, A2 and F). Association could be demonstrated only between syphilis and TcAb in the female group (p = 0.005), but the low number of donors found with this association (n = 4) renders the effect of this correlation very small. A higher prevalence of TcAb was found in older age groups, with even gender distribution (p < 0.05), however, donors aged more than 54 years also represent a minority of the donor pool (4.83%) and the detection of positive donors in this age group also has a minor preventive effect on transfusion-transmitted Chagas disease. We conclude that when infected blood donors must be detected, specific serological screening for TcAb is essential and that currently no surrogate marker can be considered for detecting T. cruzi-infected blood donors. PMID- 7483492 TI - Relative frequencies of the two O alleles of the histo-blood ABH system in different racial groups. AB - O blood group results from the absence of glycosyltransferase activity, which is due most commonly to a single nucleotide deletion in the glycosyltransferase gene. A second type of O allele resulting from three nucleotide substitutions in the glycosyltransferase gene had its frequency recently determined in a Danish population. However, its frequency among different human populations is not known. The frequencies of the two types of O alleles were determined by DNA analysis of the glycosyltransferase gene of group O individuals of three racial groups (Caucasians, blacks and Amerindians). The mean frequency of carriers of the three-base mutation group O gene among blacks and Caucasians is 4.7%; the mutation was not observed among 100 Amerindian chromosomes. The data reveal the relative frequencies of O alleles in different racial groups, and demonstrate that the origin of this variant predates racial divergence, since it is found equally among blacks and whites. PMID- 7483493 TI - A novel common Kell antigen, TOU, and its spatial relationship to other Kell antigens. AB - A 47-year-old native American (TOU) was admitted to hospital for hip surgery. His serum agglutinated all red blood cells (RBCs) tested except Ko and DTT-treated RBCs and was weakly reactive with RBCs known to have a weak expression of Kell antigens, namely Kmod, McLeod, Kp(a+b-) (KEL:3,-4) and K:-13 (KEL:-13) phenotypes. RBCs from three siblings, a son and a daughter were incompatible with TOU's antibody. TOU's RBCs had the common Kell phenotype: K-k+Kp(a-b+c-)Ku+Js(a b+)Ul(a-)K:11,-17K:14,-24K:12,13,18,19, 22,-23(KEL:-1,2,-3,4,5,-6,7, 10,11,12,13,14,-17,18,19,-21,22,-23,-24). Since TOU's RBCs were not agglutinated by an unidentified Kell-related antibody (IAN), tests were performed to show that TOU and IAN were mutually compatible. IAN is a Latino female hospitalised for a hysterectomy. The TOU antigen was shown to be located on the Kell glycoprotein by a monoclonal antibody immobilisation of erythrocyte antigen (MAIEA) assay. The unique pattern of reactivity obtained with TOU and IAN antibodies using this assay indicated the TOU epitope to be in an area remote from other Kell antigens, namely K, k, Kpa, Kpb, Kpc, Ku, Jsa, Jsb, U1a, K11, K12, K13, K14, Wka, K18, K19, K22 and K24 (KEL1, KEL2, KEL3, KEL4, KEL5, KEL6, KEL7, KEL11, KEL12, KEL13, KEL14, KEL17, KEL18, KEL4, KEL5, KEL6, KEL7, KEL11, KEL12, KEL13, KEL14, KEL17, KEL18, KEL19, KEL21, KEL22 and KEL24) but close to the low-incidence antigen K23 (KEL23).(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7483494 TI - Laboratory procedures for the prediction of the severity of haemolytic disease of the newborn. PMID- 7483495 TI - DNA recommendations--1994 report concerning further recommendations of the DNA Commission of the ISFH regarding PCR-based polymorphisms in STR (short tandem repeat) systems. PMID- 7483496 TI - Prospective study of post-transfusion hepatitis in north London: improved sensitivity of enhanced assays. PMID- 7483497 TI - Analysing the performance of anti-HCV supplemental tests in German blood donors. PMID- 7483498 TI - Anti-HCV positivity among blood donor population from Pune, India (1981-1994) PMID- 7483499 TI - New tools for HCV confirmation. PMID- 7483500 TI - Look back on hepatitis C-virus infections of HCV-RIBA-2-positive blood donors and their respective recipients. PMID- 7483501 TI - Ultracentrifugal analysis of factor VIII and von Willebrand factor in therapeutic preparations. AB - Plasma and therapeutic preparations of factor VIII (1 recombinant factor VIII and two monoclonally purified plasma-derived factor VIII preparations, Kogenate, and AHF-M and Monoclate, respectively) were centrifuged in a sucrose density gradient, and the fractions were analyzed for factor VIII and von Willebrand factor (vWF). The residual vWF in the monoclonally purified factor VIII preparations sediments more slowly than the vWF of plasma. In the absence of added vWF, the factor VIII in all preparations sediments more slowly than plasma factor VIII. These same preparations of factor VIII added to hemophilic plasma as a source of vWF sediment differently. The addition of either recombinant factor VIII or AHF-M results in sedimentation of the factor VIII with the plasma vFW and in a position indistinguishable from factor VIII in plasma. In contrast, when Monoclate is added to hemophilic plasma in vitro, the factor VIII sediments more slowly than the vWF of the hemophilic plasma. However, 5 min after the infusion of Monoclate into a patient with hemophilia A, the factor VIII sediments with the plasma vWF. These results indicate that the addition of recombinant factor VIII and AHF-M results in random binding to all vWF multimers of plasma, while there is little exchange between the added factor VIII in Monoclate and the plasma vWF in vitro. In contrast, when the Monoclate is infused, there is rapid binding of factor VIII to the plasma vWF.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7483502 TI - Vaccination against hepatitis A virus in French hemophilic children. PMID- 7483503 TI - Value of virus filtration as a method for improving the safety of plasma products. PMID- 7483504 TI - Is mycosis fungoides associated with HTLV-I? PMID- 7483505 TI - Can C1 esterase inhibitor concentrate be a cause of the exacerbation of hereditary angioneurotic oedema? PMID- 7483506 TI - Incidence of delayed hemolytic transfusion reactions. PMID- 7483507 TI - Apparent relationship between vacuolization of fetal erythrocytes and anemia in red cell alloimmunization. PMID- 7483508 TI - [The prevalence of secondary resistance to peroral hypoglycemic agents among patients with type-2 diabetes mellitus]. AB - Prevalence of secondary resistance to peroral sugar-reducing agents (PSRA) among patients with type II diabetes mellitus was studied. The majority of patients (81%) who were allocated to insulin treatment for the first time had received peroral sugar-reducing preparation prior to that treatment. Mean duration of illness in these patients was 9.94 +/- 0.6 years. Females predominated among those persons showing resistance to PSRA; they all presented with obesity. PMID- 7483510 TI - [The function of the peripheral blood lymphocytes in patients with autoimmune thyroiditis]. PMID- 7483509 TI - [The function of the blood neutrophils in diabetics]. AB - Studies made in 54 patients and "in vitro" experiment permitted finding out that insulin exerts major effect on the blood neutrophil functional status. It activates phagocytizing function of leucocytes as a result of intensification of glycolysis and cellular respiration, favours intracellular plastic processes. Elimination of hyperglycemia, insulin deficiency are prerequisites of success in correction of disturbances in phagocytizing function of cells in patients with diabetes mellitus. PMID- 7483511 TI - [The mass of the myocardium and tolerance for physical exercise in the postinfarct period in hypertension patients]. AB - As many as 56 patients with stage-II hypertensive disease (HD) as per the WHO classification were examined. On day 30 of myocardial infarction (MI) the left ventricular myocardium mass was calculated and six months later exercise tolerance was evaluated using bicycle ergometer. The evaluation done showed that the presence of hypertrophy of the Left ventricle in HD patients by day 30 of MI was a prognostic sign of low exercise tolerance in postinfarction period, which fact permits exercise tolerability to be foreseen, and issues related to the patients' fitness for work to be settled as early as the hospital stage of rehabilitation of MI patients. PMID- 7483512 TI - [The late results of electrical cardioversion in patients with a chronic form of atrial fibrillation]. AB - The paper presents results of study of the electric cardioversion (ECV) long-term effects in 216 patients with a chronic form of cardiac fibrillation (CF). The CF relapses were found to occur in 45.4% of cases, with the 43.9% of the whole number of the above relapses falling on the first two weeks, during six months after successful ECV. CF relapses were significantly more common in patients with rheumatic and ischemic heart diseases, as well as in those persons with CF duration of more than one year and in recurrent arrhythmias in instances of left ventricle sizes exceeding 45 mm. CF form does not appear to have a demonstrable effect on the ECV long-term results. PMID- 7483513 TI - [The microflora of the gastric mucosa and its role in the development of recurrent peptic ulcer]. PMID- 7483514 TI - [Complications of the biliary system during therapeutic fasting]. PMID- 7483515 TI - [Apitherapy in the combined treatment of patients with pulmonary tuberculosis taking into account the hypophyseal-adrenal system indices]. AB - Apitherapy (Venom of bees and apiculture products) was included into combined treatment of 93 in-patients with pulmonary tuberculosis. Apitherapy had a beneficial effect on the organism of tuberculosis patients, manifested by enhancement of the treatment effectiveness and normalization of indices of endocrine system. It is recommended that the instruction on apitoxinotherapy be amended, in particular, by substantially supplementing the paragraph with indications and contraindications for giving it in active tuberculosis. PMID- 7483516 TI - [The cerebral blood flow in smokers]. AB - Effect of smoking on cerebral blood flow was studied with the aid of rheography in 60 young smokers and 30 nonsmokers (control group). Tobacco smoking was found to results in rising the tone of cerebral arteries of large and medium calibres, the pulse organ filling being on the decrease. The tone of small arterial vessels and venous network decreases. The effect of the inhaled tobacco smoke on cerebral blood flow is particularly pronounced in prolonged and continuous users of tobacco. PMID- 7483517 TI - [The clinical and epidemiological characteristics of adenovirus infection in adults]. PMID- 7483519 TI - [Broncholytic agents in the treatment of bronchial asthma]. PMID- 7483518 TI - [Immunoglobulin E and beta 2-microglobulin in the blood serum and synovial fluid of patients with rheumatoid arthritis, arthrosis deformans and traumatic arthritis]. PMID- 7483520 TI - [Immune and microcirculatory disorders in patients with a protracted course of viral hepatitis B and their correction]. AB - Immunity status and that of microhemodynamics were evaluated in 85 patients with a protracted course of viral hepatitis B (VHB). Clearcut correlation was found between the values for immunity and microcirculation. Use of parmidine and quercetin in a combined treatment of patients makes for rapid clearing of the disturbances revealed, promoting reconvalescence, reducing the frequency of VHB recurrences and chronization of pathological processes in the liver. PMID- 7483522 TI - [Psychopharmacological correction in the combined treatment of patients with vibration-induced disease]. PMID- 7483521 TI - [The prophylactic use of pectin in chronic lead exposure in industry]. AB - An effect was studied of lead on certain physiologic, hematological and biochemical parameters in workers engaged in the lead-acid cell manufacture, as was a prophylactic action of pectin in the aforementioned groups of persons. Elimination of lead from the organism was found to be on the increase after the intake of pectin-vitamin preparation (PVP, 3-4 g daily for one month), with the above parameters returning to normal. It is recommended that regular courses of pectin prophylaxis be delivered under control of adequate tests and levels of lead in indicator biological media if we are to achieve a maximal effect in prevention of chronic lead poisoning. PMID- 7483523 TI - [The working conditions of smokers and their assistants on drilling rigs during geological prospecting operations and the measures for their improvement]. PMID- 7483524 TI - [The clinical course of the early neonatal period in the newborn infants of mothers with diabetes mellitus]. PMID- 7483525 TI - [The treatment of prostatic adenoma by transurethral microwave therapy]. AB - Patients with adenoma of the prostate appear to benefit from application of the unit "Prostatron" in one session without anesthesiologic maintenance and under outpatient conditions in the absolute majority of cases, as evidenced by reports from abroad and our own experience gained in the treatment of patients with adenoma of the prostate. This permits it to be recommended for widespread use in urological settings. Clinical, uroflowmetric and ultrasonic monitoring of the urogenital system status in patients with adenoma of the prostate, which is to be delivered in all such patients,--if used in combination with TUMWT,--becomes a means of really active assessment of treatment effectiveness rather than a passive recording of the moment of worsening of the patient' state. PMID- 7483526 TI - [The effect of natural immunomodulators on the lymphocytic natural killer activity in patients with malignant lymphoproliferative diseases]. AB - The conducted investigations showed an apparent immunomodulating action of the studied natural preparations on natural resistance of patients with malignant lymphomas. The modulation effect was the greatest in patients taking esberitox during remission and while receiving radiotherapy. Administration of Ney Tumorin to patients with lymphogranulomatosis during remission appeared to be associated with substantial increase in the content of large granular lymphocytes and level of natural killer activity (almost two--fold) immediately after the end of the treatment and this tendency was to be still seen one--two months later. Pirogenal had marked stimulatory effect on natural resistance of patients with granulomatosis in the postradiation period. The studied preparations were not associated with side effects. General condition improved after their administration, as per subjective experience. PMID- 7483527 TI - [A case of histiocytosis X]. PMID- 7483528 TI - [An automated system for public opinion polling on life style, health status and health behavior]. PMID- 7483529 TI - [The efficacy of the dispensary observation of patients with ischemic heart disease who live in a rural locality]. AB - 30 patients with chronic ischemic heart disease allocated to controlled treatment and close monitoring, and 30 patients with ischemic heart disease placed under routine dispensary observation, living in rural areas, were examined. Effectiveness of dispensary follow-up of the IHD patients was found to be dependent upon three factors: use of differentiated approach to prescription of drugs, continuity of work in organization of treatment and feasibility of giving therapy under carefully health monitored conditions. PMID- 7483530 TI - [The organization and experience of the treatment of urological patients at a day hospital]. PMID- 7483531 TI - [A comprehensive treatment and rehabilitation program for drug addicts at the Dnepropetrovsk Province Drug Abuse Clinic]. AB - On the basis of the experience gained in the treatment of patients who are drug addicts a complex treatment-and-rehabilitation programme has been developed and adopted. It is a psychotherapeutically--oriented programme that includes the following steps: selection of patients, promotion of health of the family, staging, participation of social workers--former patients. The end objective is to achieve a lasting remission in addicted patients, promotion of health of their relatives and normalization of family relations. Such an approach to solving the problem differs from nonmedicamentous methods widely employed now in the Ukraine and UIS republics, which have not been associated with demonstrable effects in treatment and, more important, rehabilitation of addicted persons. PMID- 7483532 TI - [Latent anaerobic infection in the development of pathological processes in the oral cavity]. PMID- 7483533 TI - [The scientific, pedagogical and medical activities of Prof. V. P. Rudin]. PMID- 7483534 TI - [M. A. Bulgakov and questions of medical deontology]. PMID- 7483535 TI - [The participation of the arteriolar endothelium in regulating renin secretion]. PMID- 7483536 TI - [The treatment of distal neuropathy--an important means for preventing the development of diabetic gangrene]. PMID- 7483537 TI - [The intraperitoneal chemotherapy of patients with ovarian cancer]. PMID- 7483538 TI - [The scientific information culture of the physician]. PMID- 7483539 TI - [The results of a study of disability among those who participated in the cleanup of the aftereffects of the accident at the Chernobyl Atomic Electric Power Station (based on data from the Ukrainian National Registry)]. PMID- 7483540 TI - [The correction of disorders in free-radical lipid oxidation in the blood of persons working in an area with elevated ionizing radiation]. PMID- 7483541 TI - [A case of prolonged hematopoietic depression after acute radiation sickness in a man who worked in the cleanup of the aftereffects of the accident at the Chernobyl Atomic Electric Power Station]. PMID- 7483542 TI - [The histochemical and histoenzymological changes in the gastric mucosa and the state of the connective tissue in peptic ulcer patients during dynamic rehabilitative treatment]. AB - Combined differentiated rehabilitative and staged treatment of 167 patients with peptic ulcer resulted in stable reduction in cellular membrane permeability, subsiding of catabolic processes in cells, steady normalization of enzymic activity of tissue respiration, as well as activation of reparation processes in connective tissue elements and structures located in the connective tissue (capillaries included). The described mode of treatment appeared to be superior to the conventional one, resulting in a substantial improvement in microcirculation in gastric mucosa. Normalization of protein-synthetic processes in the connective tissue makes for enhancing of the gastric mucosa resistance and prevention of the relapse of the disease. PMID- 7483543 TI - [The histoenzymological and ultrastructural changes in the gastric mucosa in peptic ulcer after treatment with pirenzepine and histamine H2-receptor blockaders]. AB - A study made in 72 patients showed a positive influence in ulcer disease of gastrocepine and H2-blockers of receptors to histamine on histoenzymological and ultrastructural changes in the mucous membrane of the stomach and duodenum. Gastrocepine was found to be superior to H2-blockers of receptors to histamine due to its apparent cytoprotective effect and ability to improve microcirculation. PMID- 7483544 TI - [The functional and sonographic parallels in chronic pancreatitis]. AB - Ultrasonic histogramme was studied in 35 patients with chronic pancreatitis and 20 donors, taken in the area of the head and body of the pancreas, as were levels of immunoreactive trypsin--fasting and after standard food load,--and C-peptide in blood serum on an empty stomach. To increase informative value and specificity of ultrasonic and radioimmunologic investigations an index has been suggested, based on the findings from histogramme and radioimmunity analysis. PMID- 7483545 TI - [The effect of thymalin, dalargin and "Mucosalin" on the course of an experimental duodenal ulcer]. AB - A peptide was obtained at our chair, from the mucous membrane of the stomach of the pig, using a method of acid extraction. The above peptide belongs to the group of cytomedines and is conventionally called "mucozaline". As many as 80 Wistar male rats were used in comparative evaluation of thymaline, dallargine and "mucozaline" for their effect on the course of duodenal ulcer in the rat. The results obtained showed that regulatory peptides play an important part in protection of the gastric and duodenal mucosa against the damaging factors. Of the above agents, it is "mucozaline" that has the greatest protective action, with dallargine and thymaline following it in the declining order. PMID- 7483546 TI - [An evaluation of the efficacy of the new Ukrainian preparation Erbisol in patients with chronic hepatitis]. AB - A new drug preparation "Erbisol" was studied for its effect in the treatment of chronic hepatitis. There were 52 patients; of these, chronic active hepatitis was diagnosed in ten, persistent hepatitis in 38, cholangiolytic hepatitis in 4 patients. The preparation has a beneficial effect on the clinical course of the disease and makes for the normalization of biochemical and immunological indicators in patients with persistent hepatitis and in those with chronic active hepatitis with mild degree of activity. In patients with cholangiolytic hepatitis "Erbisol" should be used with caution. The studies made allowed for the conclusion to be drawn to the effect that the preparation "Erbisol" is a highly effective hepatoprotective and immunomodulating drug. PMID- 7483547 TI - [Enteroinsular hormones in chronic gastritis with secretory insufficiency]. AB - The blood serum content of secretin, vasointestinal peptide (VIP), glucagon and insulin was studied in 112 patients with chronic gastritis presenting with secretory deficiency, and 12 essentially healthy subjects. It was established that in surface, fundal and pangastritis the levels of secretin and VIP did not differ from the values in healthy individuals. The secretin concentration tended to be the lowest in atrophic fundal and atrophic pangastritis. They also had significantly elevated concentrations of VIP and glucagon, with insulin decreased. Caused by different exo- and endogenous factors phenomena characteristic of gastritis, such as disturbances in acid-producing function of the stomach, to mention but one of its manifestations, result in changes of stimulatory effects of acidification on enterinic elements of the small intestine, which circumstance leads to discoordination in output of intestinal and pancreatic hormones particularly pronounced in atrophic gastritis. PMID- 7483548 TI - [The pathogenesis of myocardial infarct in young people]. PMID- 7483549 TI - [The prospects for using a low-energy exposure of the erythrocytes during cryopreservation]. PMID- 7483550 TI - [A new method for treating patients with chronic prostatitis]. AB - A new preparation is reported for treatment of chronic inflammation of the prostate, which substantially lowers rates of patients' complaints and depresses the secretion leucocyte reaction. Prostatilen was shown to be capable of normalization of immunity status of the chronic prostatitis patients thus lowering the microbial index of the cultured prostate secretion microorganisms. The following new nonantibacterial strategy of treatment of chronic prostatitis patients is proposed: prostatilen given as a single agent or in combination with immunomodulators and physiotherapeutic methods. Antibacterial therapy is to be instituted on a short-term basis only during the period of exacerbation of the inflammatory process. PMID- 7483552 TI - [The correction of the metabolic and microcirculatory disorders in patients with atherosclerotic and involutional depressions]. AB - A total of 42 patients with atherosclerotic--and 34 with involutional depression were studied by conjunctival biomicroscopy for particular features of lipid peroxidation and state of microcirculation before and after treatment with vasoactive preparations, antioxidants and calcium antagonists. POL were found to be increased in patients with atherosclerotic depression irrespective of the syndrome, and in involutional depression with anxious symptomatology, as well as in those patients in stuporous state, this being accounted by a decreased provision of the organism with antioxidants as well as by an inadequate activity of antioxidant enzymes; high incidence rate of intravascular and vascular changes was also noted in the above patient populations. Tocoferolum acetatum and corinfar potentiated antiaggregational and spasmolytic actions of vasoactive preparations, which fact made for an earlier eradication of microcirculatory disturbances, decrease in the free radical oxidation activity and recovery of those patients getting out of the psychotic state. PMID- 7483553 TI - [The characteristics of the functional interrelationship of the immune and fibrinolytic systems in psoriasis patients]. AB - A total of 83 patients with psoriasis were examined; of these, 53 individuals had disseminated forms of psoriasis (group 1), 30--generalized or global psoriatic erythroderma (group II). Studied in the peripheral blood were activity of interleukins--1 and 2 (IL-1 and 2), concentration of plasmic fibronektin and relative count of lymphocytes expressing receptors to urokinase. There have been revealed in group I patients moderate increase in IL-1 activity, apparent lowering of IL-2, reduction in concentration of plasmic fibronektin, decrease in relative count of lymphocytes expressing receptors to urokinase in both clinical groups. Negative correlation was established between the latter two parameters, which was the greatest in patients with psoriatic erythrodermia. PMID- 7483551 TI - [The indices of the antioxidant system and the status of the cerebral blood supply in patients with an ischemic stroke on apitherapy]. AB - It has been established that the use of apitherapy (pollen and propolis) to treat patients with ischemic insults leads to deeper positive shifts in indices of the antioxidant system and brain blood supply. This, in its turn, makes for rapid and complete restoration of disturbed and lost functions of the patients' organism. PMID- 7483555 TI - [The effect of seasonal biorhythms on the clinical course of hepatitis B]. PMID- 7483554 TI - [The antioxidant correction of cytogenetic disorders in viral hepatitis]. AB - It has been established that in the acute period of viral hepatitis (VH) the level of chromosome aberrations is 4.4-fold and 6.6-fold that of the norm in VHA and VHB respectively, with the chromatid type aberrations predominating. Frequency of metaphases with chromosome aberrations in substantially higher in patients selected from those categories of individuals working at chemical enterprises and persons living in the zone of exposure to discharges from huge metallurgical works and coking plants. There is a correlation between the markedness of cytogenetic disturbances and degree of cellular immune deficiency. Use of antioxidants (tocopheroli acetas, parmidin, quercetin) in the complex of drug preparations contributes to the normalization of cytogenetic indices and reduction of frequency of protracted and chronic forms of hepatitis. PMID- 7483556 TI - [The count of large granular lymphocytes in the blood and natural killing in tuberculosis]. AB - Blood content of large granular lymphocytes (LGL) and natural killing were studied in patients with pulmonary tuberculosis and in the time course of experimental tuberculosis in CBA mice. Evolution of tuberculosis was found to be accompanied by lowering of blood concentration of LGL and a tendency toward enhancing the natural killing more pronounced in severe course of the illness presenting with T-cellular immunologic deficiency, lowered tuberculin sensitivity of T-lymphocytes and high titres of antituberculous antibodies. The revealed changes are of compensatory character. PMID- 7483557 TI - [The morphofunctional changes in the ventricular myocardium in experimental infarct and its treatment by hyperbaric oxygenation and tocopherol acetate]. AB - Experimental myocardial infarction was induced in 50 rabbits, 25 of which were subjected to hyperbaric oxygenation (HBO) used as a single procedure or in combination with the antioxidant tocopheroli acetas. HBO was found to promote reduction of the myocardial lesion focus and enhancement of the regeneration processes but in everyday employment over long periods of time it causes dystrophic changes in the remote zones of the myocardium. Administration of tocopheroli acetas before the HBO sessions allows the toxic action of oxygen to be safely prevented. PMID- 7483558 TI - [The carcinogenic risk factor in the job of a steel welder]. AB - The paper presents results of evaluation of carcinogenic risk in welders with due emphasis to sensibilization to organ-specific tumorous antigen of the lung determined by a test for electrophoretic mobility of indicator cells. Relationship was found between frequency and markedness of sensibilization and blood serum content of iron. It is suggested that carcinogenic risk in the occupation of the welder might be associated with both exposure to chromium, nickel, asbestos endowed with blastomogenic properties, and an increase accumulation of iron in the organism. PMID- 7483559 TI - [The characteristics of the biological action and regulation of submicroscopic aerosols in the industrial environment]. AB - Biologic action was studied of the silicon nitride powder with varying disperse composition. When producing and using the silicon nitride powder on an enlarged scale one must remember of its higher toxicity and fibrogenicity. There is a need of employing some method of determination of the submicroscopic aerosol disperse composition under industrial conditions. PMID- 7483560 TI - [The effect of lead compounds on the development of arterial hypertension in workers in the radio electronics industry]. AB - As many as 2723 workmen aged 15-64 years (1736 women, 987 men) from a radio electronic industry enterprise were examined with the purpose of finding out the prevalence of arterial hypertension (including borderline and persistent ones), depending on the length of service, in two different occupational groups: the basic one consisting of those exposed to lead compounds occupationally, and control one. The examination showed that the level of dynamics of the AH patient relative numbers was significantly higher in those contingents of workmen who had been occupationally exposed to lead compounds for long periods of time. PMID- 7483561 TI - [The health of women engaged in agricultural production in Ukraine (the experience and perspectives of epidemiological research)]. PMID- 7483562 TI - [Penetration of neurotropic viruses into the central nervous system after airborne mode of transmission]. PMID- 7483563 TI - [Comparative study of the nucleotide sequence of the gene for the P/C wild strain or L-16 vaccine strain of measles virus]. AB - Nucleotide sequences of P/C gene of measles virus IL wild strain and L-16 vaccine strain were compared with the previously reported nucleotide sequence of Edmonston vaccine strain P/C gene. The primary structure of IL strain P/C gene was characterized by two substitutes in 395 (U-C) and 722 (G-A) positions in comparison with the primary structure of a similar Edmonston strain. Nucleotide sequence of L-16 vaccine strain P/C gene differed from the similar sequence of Edmonston vaccine strain by 3 substitutes in 90 (A-G), 399 (U-C), and 448 (U-C) positions. Comparative analysis of nucleotide substitutes in the P/C gene of L 16, IL, and Edmonston strains indicates that the mutations revealed in the L-16 strain P/C gene are not caused by the strain attenuation, but merely reflect the difference between the strains. PMID- 7483564 TI - [Protective activity of vaccinia virus envelope proteins isolated using nonionic detergents]. AB - Several detergents and chemical compounds--Tweens 40, 60, 80, Triton X-100, Triton WR-1340, polyvinylpyrrolidone, dimethylsulfoxide, urea, n-butyl, MESK, and combinations thereof were used for the isolation of surface proteins of vaccinia virus. Optimal conditions for the treatment of the virus with detergents were selected, permitting isolation of vaccinia virus surface proteins p35 and p61. Mouse experiments yielded data on the protective properties of the isolated proteins. Protein p35 may turn to be one of the major proteins responsible for the formation of protective immunity in vaccination with vaccinia virus. PMID- 7483565 TI - [Certain pathogenetic characteristics of a disease in monkeys in infected with the Marburg virus by an airborne route]. AB - Time course of Marburg virus (strain Popp) accumulation and changes in hematological parameters were studied in aerosol infected M.rhesus monkeys. The lungs were the first organ in which the virus was detected after respiratory infection of monkeys. Four days after inoculation the virus was detected in the liver, spleen, blood, and thymus. Six days after inoculation the virus was present in virtually all organs and secretions. The period of fever was associated with manifest leukopenia in primates. Blood clotting time drastically increased by the moment of animal death. PMID- 7483566 TI - [A promising method for preparative production and purification of the Marburg virus]. AB - A method for the production of Marburg virus in preparative amounts has been developed. It is based on sedimentation of the virus from blood plasma of infected guinea pigs by ultracentrifugation followed by purification in sucrose density gradient or gel chromatography on macroporous glass sorbents. The optimal terms of blood collection in infected animals were determined. Purified virus did not lose its biological activity. Concentrated virus preparations were studied by electron microscopy and polyacrylamide gel electrophoresis. PMID- 7483567 TI - [Pathogenetic criteria for evaluating the virulence of tick-borne encephalitis strains isolated in the southern part of the Far East]. AB - A comparative virological, immunological, and pathohistological study of 5 tick borne encephalitis (TBE) strains differing by virulence was carried out in golden hamsters. The authors characterize the TBE virus pathogenicity sign, denoted as Pmaic genetic marker (pathogenicity for hamsters at intracerebral inoculation), which helps understand the essence of immunopathogenetic mechanisms of the TBE infectious process in man. PMID- 7483568 TI - [Mosquito-borne arboviruses in the Baikal region]. AB - A total of 20 virus strains were isolated from arthropods and rodents in the Baikal region in 1992. Thirteen of these were California serogroup viruses, 4 Batai virus strains (Bunyaviridae, Bunyavirus), and 3 Gheta virus strains (Togaviridae, Alphavirus, Semliki complex). The rates of mosquito infection with California, Batai, and Gheta viruses were 1:6500, 1:24000, and 1:24000, respectively. The results of the neutralization test indicate the similarity of the isolated California complex strains to the mountain hare virus, the most pathogenic virus in this group. PMID- 7483570 TI - [Interferon status in predicting the outcome of acute viral hepatitis B]. AB - Analyzes the interferon status of patients with various outcomes of acute hepatitis B at different stages of the disease. Assesses the possibility of predicting cure and transformation of the acute disease into chronic hepatitis B. Enumerates factors influencing the efficacy of interferon therapy and the outcome of viral hepatitis B. PMID- 7483569 TI - [Frequency of encountering markers of viral hepatitis A, B, and C in the aboriginal inhabitants of the northwest region of Siberia]. AB - A minor population of Nenets living near the village of Samburg in the Purov District, Tyumen region was screened for serological markers of virus hepatitides A, B, and C (VHA, HVB, and HVC, respectively). Commercial kits manufactured by VectorBest JSC, Russia, were used. All the tested sera contained antiHVA IgG; IgM antibodies to VHA were detected in none of the tested sera, anti-HVC IgG were found in 1 (0.93%) serum, antiHBc IgG in 11 (10.2%), and HBsAg was detected in 2 (1.85%) serum samples. The sera positive by antiHBc IgG and HBsAg were tested for DNA by the polymerase chain reaction. HVB DNA was found in two HBsAg-positive sera with the HBsAg subtype determined as "non ad". PMID- 7483571 TI - [Humoral antiviral immunity in children with hematologic neoplasms]. AB - Antiviral humoral immunity was studied in patients with Hodgkin's disease, idiopathic thrombocytopenic purpura (ITP), and aplastic anemia (AA). Insufficiency of antiviral humoral immunity as regards antigenically unrelated viruses and Mycoplasma pneumoniae was characteristic of all these conditions. A remission of Hodgkin's disease and ITP was associated with a reliable increase of the studied parameters in comparison with the acute period of the disease. After splenectomy humoral immunity parameters were virtually unchanged in patients with ITP, whereas in patients with Hodgkin's disease and AA the deficiency of antibody production is 40% increased after the operation. PMID- 7483572 TI - [Natural killers and cytotoxic lymphocytes in classical hog cholera]. AB - The formation of immune mechanisms directed at elimination of infected cells and including the activity of natural killers and cytotoxic lymphocytes was assessed in pigs infected with hog cholera virus. In acute disease natural killer activity in the blood is reduced, while in vaccinal process it is increased. Vaccination in parallel with cyclophosphamide immunodepression lead to inhibition of natural killer activity. Leukocytes and lymphocytes of immunized pigs can cause cytolysis of autologous targets infected with hog cholera virus. PMID- 7483573 TI - [Natural virus carrier status in the most prevalent species of forest insects- phytophages and its connection with host viability]. AB - Virus carriership of Ocneria dispar L. and Ocneria monacha L. was studied by microscopy and nucleic acid hybridization. A correlation was revealed between virus infection rate and viability of the insects. A high level of virus carriership in insects did not always lead to their death and viability deterioration. PMID- 7483574 TI - [Modern approaches in the struggle against rubella in the European region]. PMID- 7483575 TI - [Strains of cultured cells for biotechnology and scientific research]. PMID- 7483576 TI - [Physicochemical properties of a beta-glucan from Sclerotinia sclerotiorum]. AB - Sclerotan (SSG) was an extracellular polysaccharide from Sclerotinia sclerotiorum by submerged fermentation. It had potential immunomodulating and antitumor activity. The SSG was a glucan composed of beta-linked D-glucoses. It was hard to dissolve in water under normal condition, but its aqueous solution had fine rheological properties. Its intrinsic viscosity [eta] hardly changed with ionic strenth. Change of its [eta] value was not remarkable between pH 1.88-12.36. Nevertheless, when the pH came to 13.32, the [eta] value decreased rapidly due to change of molecules conformation. Effect of temperature < or = 90 degrees C and heat treatment on apparent viscosity of SSG solution was minor. PMID- 7483577 TI - [Isolation and identification of a pathogen of grasshoppers]. AB - A pathogen was isolated from natural dead pests of Ceracris kiangsu in Geleshan farm of Chongqin. Its pathogenecity was confirmed by the law of KOCK. It was identified as Pseudomonas pseudoalcaligenes according to its physiological and biochemical properties as well as the G + C content of DNA (63.73mol%). The results of preliminary bioassay show that the pathogen can infect the grasshoppers and Ceracris kiangsu, and also can infect other pests of grassland in a certain extent. PMID- 7483578 TI - [Isolation and identification of Rhodopseudomonas viridis and Rhodopseudomonas sulfoviridis]. AB - Under selective culture conditions, two pure cultures (strain G and strain SG) of the purple nonsulfur photosynthetic bacteria containing bacteriochlorophyll b were isolated from pulp effluent and sludge of sewage treatment plant by means of agar shake dilution. Although the cells of strain G and strain SG contained bacteriochlorophyll b and carotenoids, intra-cytoplasmic membranes were of lamellar type parallel to cytoplasmic membrane, the two strains differ in using various organic carbon sources, depending on reduced sulfur compounds and assimilate sulfate. According to Bergey's Manual of Systematic Bacteriology Volume 3, strain G was identified to be Rhodopseudomonas viridis,, strain SG was identified to be Rhodopseudomonas sulfoviridis. PMID- 7483579 TI - [The phenoloxidase test and its application to identify Cryptococcus neoformans with various biological characteristics]. AB - Cryptococcus neoformans strains with various biological characteristics were examinated the phenoloxidase activity on the caffeic acid cornmeal agar (CACA). Other medically important yeasts were also included in this research. Firstly, thirteen reference strains of Cryptococcus neoformans were confirmly found positive phenoloxidase production on the CACA medium. Then, in total 150 isolates of yeasts, forty three Cryptococcus neoformans strains were phenoloxidase positive, other than that 107 other yeast strains were all negative. It is suggested that Cryptococcus neoformans strains with different biological features specifically and uniquely produce phenoloxidase, thus the phenoloxidase test can be applied as a useful tool to identify this pathogenic yeast. Our results also show that the culture on the CACA medium is an effective method to test the phenoloxidase activity within 72 hours. It is further approved that the urease test will not be able to be a screening test for Cryptococcus neoformans in the clinical laboratory, but it is still valuable to identify urease negative Cryptococcus neoformans when the test is used in combination with the phenoloxidase test. PMID- 7483580 TI - [Cloning and sequencing of ntrBC genes from Azospirillum brasilense]. AB - A gene library of Azospirillum brasilense Yu62 was constructed in EMBL3. The library was screened with PCR amplified fragment as a special probe. Ten positive plaques (EA1-EA10) were selected. Detection results showed they contained two different types of clones, representing as EA4 and EA9 respectively. Southern hybridization of EA4 displayed that target gene was located in a 2.9kb EcoRI fragment. Sequence of this fragment had allowed the position and identification of ntrC gene, which encoding a protein of 53469, consisted of 480 amino acids. In the upstream of ntrC, a complete ntrB coding region was also found, which encoding a protein of 43487, consisted of 400 amino acids. Homologous analysis of the deduced amino acid sequences of ntrC and ntrB from different bacteria demonstrated that A. brasilense was closer to Rhizobia than to other free-living diazotrophs. PMID- 7483581 TI - [Induced expression of whiG, a gene crucial for sporulation of Streptomyces coelicolor]. AB - whiG gene has been subcloned into Streptomyces expression vector pAK203 containing inducible promoter tipA. The expression of whiG gene promoted the spore formation of S. coelicolor J1501 and recovered the sporulation ability of whiG-deficient S. coelicolor C71. Increased amount of whiG gene product was detected by Western blot hybridization after induction of thiostrepton. It will be helpful for the future study of in vitro transcription of whiG-dependent promoters. PMID- 7483582 TI - [The distribution of SbV nonstructural protein 2 (nsP2) in host cell]. AB - The distribution of SbV nsP2 has been observed by means of immunolabelling electron microscopy. The results showed that SbV nsP2 existed both in the cytoplasm and in the nucleus of host cell. When the cDNA of SbV nsP2 was transfected in host cell, the nsP2 also accumulates in the nucleus. PMID- 7483583 TI - [Studies on fused recombination of Gibberella fujikuroi--gibberellin-producing strain]. AB - A pair of nutrient complementary auxotrophic mutants were used as fusion parents. Their protoplasts were prepared enzymatically and PEG4000 was used as fusion agent. The prototrophic recombinants were selected directly on the minimal regeneration medium. The recombination frequency was about 10(-7). Some unstable heterocaryons were occurred and the frequency was about 10(-5)-10(-6). Mutation of pigment, hyphae morphology and gibberellin production were induced through the fused recombination. The positive mutation, negative mutation of gibberellin yield among the recombinants were 15.3% and 53.1% respectively. Gibberellin yields of recombinants signed RN2 and RG14 were over 25% than that of the prototrophic parent 207 strain. PMID- 7483584 TI - [Synthesis of S(+)-2-(4-isobutylphenyl) propionic acid by asymmetric hydrolysis of microbial enzyme. II. Reaction conditions and product extraction]. AB - Trichosporon cutaneum 158 exhibited higher sterospecificity to hydrolyze substrate ibuprofen ester. Among the five chosen substrate, asymmetric hydrolysis of methyl ester or isopropyl ester formed S(+)-ibuprofen of 97% ee. While ethyl ester not only gave ee 93% but also showed the highest hydrolytic activity. The optimum pH range for asymmetric hydrolysis was 6.5-7.0. Ability of resolusion was stabler in the range of temperature 28-37 degrees C. This yeast produced intracellular enzyme. Hydrolysis can be carried out by powered cells treated by iced-acetone to improve the specificity of resolution. The product S(+)-ibuprofen can be got with acid-base reaction and organic solvent extraction, unreacted esters can be recovered simultaneously. PMID- 7483586 TI - [The studies on the effect of proliferative response of peripheral blood lymphocyte of healthy persons by preparations of M. vaccea]. AB - This paper present that preparations of M. vaccea, which included suspensions of live bacilli, irradiation-killed bacilli and high temperature killed bacilli, for the effect on lymphocyte proliferative response of peripheral blood in healthy persons were compared by 3H-TdR incorporation method. CPM of control was 448 +/- 131, CPM adding suspensions of live M. vaccea, irradiation-killed bacilli and high temperature killed bacilli were 1037 +/- 194, 2299 +/- 140, 1819 +/- 528, 994 +/- 186, respectively. Stimulative index of all four preparations were more than 2.0, specially suspensions of live and irradiation killed M. vaccea is more effective than BCG and high temperature killed M. vaccea, which were 5.13, 4.06, respectively. In another experiment, suspensions of live, irradiation-killed and high temperature killed M. vaccea plus rIL-2 were found more effective than rIL-2 alone. CPM of them were 9544 +/- 1727, 8530 +/- 714, 8230 +/- 1035, and 3721 +/- 1336, respectively, but BCG was 6904 +/- 1218. The results show that preparations of M. vaccea tested have distinctly promotive effect for proliferative response of lymphocyte in vitro. The addition of rIL-2 to suspendions of live M. vaccea, irradiation killed and high temperature killed M. vaccea cooperatively increased the effects of proliferative response mentioned above. PMID- 7483587 TI - [The study on the simplified agar media for mycobacteria]. AB - Three kinds of simplified agar media were reported in this paper, which are easy to prepare and are simple in constituents. The velocity and the quantity of growth of seven strains tested on simplified agar media 309A and 309C were similar to or prevail over those on Lowenstein-Jensen medium and No. 92 potato soup agar medium. Simplified agar media 309C and 309A could be applied to isolate mycobacteria from sputum and to test the drug susceptibility of M. tuberculosis. PMID- 7483585 TI - [A new zoonosis--investigation of Gardnerella vaginalis disease of fox. IV. Study of dot-ELISA for the detection of serum antibody of Gardnerella vaginalis disease of fox]. AB - 192 Gardnerella vaginalis antibody positive sera from artifically infected foxes, immunized foxes and foxes that the bacterial examination were positive were analysed by the established Dot-ELISA. The results showed that 187/192 were positive, the positive rate was 97.3%. Block and cross tests showed that the method was specific. The method was 28.1 times sensitive as PAT and 5.5 times sensitive as MAT respectively. The repeatability of the test was good and the positive coincidence rate among the three techniques was 100%. PMID- 7483588 TI - [Characters and insecticidal polypeptide of a new strain of Bacillus thuringiensis subsp. kenyae in China]. AB - A new strain of Bacillus thurinigiensis Bt with high toxicity against noctuidae larvae has been screened for many generations since isolated from larvae candle of Aphomia gularis in Yiyang County, Jiangxi Province, in China. By comparison and analysis of results of physiological and biochemical test, flagella antigen identification and esterase type, the strain is identified as H4a-4c Bt serovar. kenyae. Since its crystal protein type and plasmid type are different from those of Bt serovar. kenyae's type 023. The strain is assigned a novel strain: Btken Ag. Btken-Ag's parasporal crystals are multi-morphorous: bipyramid, cube, small irregular sphere and embedded. After UMT dissolve, PAGE and SDS-PAGE separation preparation and analysis, it is found that its major insecticide component to Heliothis armigera is 61kD toxic protein. By ELISA homology analysis, it is found that this toxic protein has high homology with crystal protein of 023 and 7501 (H4a-4c), partly homology with that of HD-1(H3a-3b), but no homology with that of Bti(H14) and (Bacillus sphaericus) Ts-1 strain. In bioassay with larvae from Culex pipiens, Pseudaletia unipuncta and Heliothis armigera, together with other ten Bt strains, Btken-Ag is toxic to 3 star larvae of Culex pipiens; two isolates of Btken-Ag (b1-4 and H4-1) show higher toxicity than type strain 023 and HD-1 do to Pseudaletia unipuncta and Heliothis armigera. PMID- 7483589 TI - Improving identification of and intervention for alcoholism. AB - A retrospective hospital medical record review was done using 45 diagnoses or laboratory findings that are associated with alcohol abuse. The reviewer assessed the level of documentation of alcohol consumption in relation to alcohol-related disorders before and after an intervention to heighten house staff's recognition of alcoholism. Of the patients with at least 1 alcohol-related disease, 58% were explicitly asked about their consumption of alcohol. The mean number of alcohol related diseases was 3.8 +/- 2.3 in the group questioned compared with 1.9 +/- 1.4 in the group not asked (P < .01). After the intervention, 90% of patients with at least 1 alcohol-related disease were asked about alcohol consumption. Once again, those asked had an average of 3.9 diseases compared with 2.1 in the other group (P < .01). Only younger age, increased specificity of alcohol-related disease, and the promotion of physician awareness were important factors for influencing documentation. Introducing a program for detecting alcoholism can have a beneficial effect on physicians' identification of alcoholism in patients with alcohol-related illnesses. PMID- 7483590 TI - Screening for depression among newly arrived Vietnamese refugees in primary care settings. AB - A brief, culture-specific, self-report screening measure for depression, the Vietnamese Depression Scale, was used to determine the prevalence of depressive symptoms among 1,998 consecutive adult Vietnamese refugees who presented at 10 public health clinics within 2 months of their arrival in the United States. Of these patients, 6% met the criterion for a probable case of depression ("positive"). Being divorced, separated, or widowed and poorly educated were strongly associated with a greater likelihood of screening positive. Somatic complaints were common and induced considerable anxiety about physical health status. Nearly a third of the patients reported sadness and dysphoria; culture specific symptoms of depression also were prevalent. Our findings document the feasibility of screening for depression using the Vietnamese Depression Scale among Vietnamese refugees, particularly in primary care settings where they are first likely to be seen by health professionals after arrival in their host country. PMID- 7483591 TI - Role of previous claims and specialty on the effectiveness of risk-management education for office-based physicians. AB - We analyzed the medical malpractice claims data of 1,903 physicians between 1981 and 1990 to assess the efficacy--a reduced incidence of future claims and decreased payout in the event of a claim--of risk-management education for office based physicians. Physicians were participants in the Oregon Medical Association's medical liability program and represented all recognized specialties and all geographic areas of the state. Each physician's claim and payout history before and after 4 sequential risk-management education programs was entered into a random-effects probit model that allowed for a longitudinal rather than a cross-sectional analysis. For most physicians, there was increased claim vulnerability following 1 or 2 risk-management education courses but decreased vulnerability after additional courses. Among all physicians, having a previous claim substantially increased the risk for a future claim. Risk for an additional claim doubled (from 7% to 14%) for physicians who had a claim in the previous year. Of all specialists who have had claims, anesthesiologists (reduction in claims incidence from 18.8% to 9.1% and in payout from 14.6% to 5%) and obstetrician-gynecologists (reduction in claims incidence from 23.3% to 15.2% and in payout from 11.6% to 4.2%) benefit most from cumulative risk-management education. PMID- 7483592 TI - Common injuries of the shoulder. Diagnosis and treatment. AB - Shoulder pain is often the presenting complaint of patients seeing their primary care physicians. Overuse and traumatic injuries make up most of the causes. A physical examination with minimal diagnostic tests can lead to the correct diagnosis in most cases. Most conditions can be treated conservatively (nonsurgically). Appropriate referral to a specialist depends on the severity of the initial injury or the patient's lack of response to conservative treatment (or both). We discuss common injuries of the shoulder, emphasizing a practical diagnostic and therapeutic approach. PMID- 7483594 TI - Steroid-resistant asthma. PMID- 7483595 TI - Effectiveness of allergy immunotherapy for asthma. PMID- 7483593 TI - Current management of ductal carcinoma in situ. AB - Ductal carcinoma in situ represents a biologically and histologically heterogeneous group of lesions characterized by the proliferation of neoplastic epithelial cells confined to the ducts of the breast. Before screening mammography, ductal carcinoma in situ was considered uncommon; patients were usually diagnosed by a breast mass or bloody nipple discharge, and their treatment was mastectomy. Today it represents 20% to 30% of mammographically detected breast cancers and 10% to 15% of all diagnosed breast cancers in the United States. The invariable progression of this cancer to invasive breast cancer requiring mastectomy has been challenged, but because most patients have been treated with mastectomy, knowledge about ductal carcinoma in situ is limited and primarily based on retrospective data. Further insight will emerge from randomized prospective studies that are near completion. Currently available data indicate that breast-conserving treatments are valid alternatives to mastectomy for most patients with this disease. PMID- 7483597 TI - Occupational exposure to latex. PMID- 7483596 TI - Changing face of HIV/AIDS care--mother-fetal and maternal-child HIV transmission. PMID- 7483598 TI - Value of home peak flow monitoring for asthma control. PMID- 7483599 TI - Caution with inhaled corticosteroids in childhood asthma. PMID- 7483600 TI - Long-acting beta 2-agonists and their role in asthma management. PMID- 7483601 TI - Asthma and air pollution. PMID- 7483602 TI - Genetic basis of the primary immunodeficiency syndromes. PMID- 7483604 TI - Hyperammonemia with severe methanol intoxication. PMID- 7483603 TI - Necrotizing fasciitis and septic shock caused by Vibrio cholerae acquired in San Diego, California. PMID- 7483605 TI - MELAS syndrome. PMID- 7483606 TI - Inherited liver diseases in adults. AB - Important inherited disorders causing acute and chronic liver disease include hemochromatosis, Wilson's disease, alpha 1-antiprotease (antitrypsin) deficiency, and cystic fibrosis. The detection of an index case has implications for screening family members. A normal life span can be expected with treatment in asymptomatic patients with Wilson's disease and hemochromatosis. We present a clinical approach to disease recognition, investigation, and screening. PMID- 7483607 TI - A case of free air in the peritoneum. PMID- 7483608 TI - The value of the history and physical for shoulder pain. PMID- 7483609 TI - Cure, conservation, confusion, chaos. PMID- 7483610 TI - Coronary artery involvement in Kawasaki disease--diagnosis and treatment. PMID- 7483611 TI - More on the use of ultrasonography in the emergency department. PMID- 7483612 TI - More on the use of ultrasonography in the emergency department. PMID- 7483613 TI - Silicone implants in men. PMID- 7483614 TI - Congenital malaria in twins. PMID- 7483615 TI - Sarcoidosis and beryllium exposure. PMID- 7483616 TI - [Magnesium or proajmaline (neogilurytmal) in treatment of recurrences of paroxysmal supraventricular tachyarrhythmias?]. AB - In order to compare the effectiveness of magnesium with that of proajmaline in preventing of recurrences of paroxysmal tachyarrhythmias. 60 consecutive patients were studied who had been previously treated for paroxysmal arterial fibrillation (PAF) or paroxysmal supraventricular tachycardia (PSVT). For the studies the patients were qualified who had been effectively treated during acute phase with intravenous magnesium (magnesium sulphate) and ajmaline (gilurytmal). The patients were randomly divided into three groups of 20 persons in each, every group received different oral treatment: in the first group--magnesium (magnesium carbonate) 3 x 0.6, in the second--proajmaline (neogilurytmal) in dose 3 x 20 mg, and in the third--neogilurytmal 2 x 20 mg and magnesium 2 x 0.6. During three months of observation the effectiveness of treatment and incidence of adverse effects were assessed. The following was found: neogilurytmal showed higher effectiveness (60%) than magnesium (30%), while the combined treatment demonstrated similar effectiveness (65%) as neogilurytmal alone with lower drug doses and lower incidence of bad tolerance manifestations (10% vs 20%). PMID- 7483617 TI - [Liver steatosis assessed by ultrasonographic examination in patients with hyperlipoproteinemia]. AB - Ultrasonographic examinations were carried out in 174 patients with various types of hyperlipoproteinaemia, determining the incidence of the features of liver steatosis. In the group of 77 patients with hypercholesterolaemia (TCH > 200 mg%) the features of liver steatosis occurred in 13 cases (16.9%). In 90 patients with mixed hyperlipidaemia (TCH > 200 mg%, TG > 2.3 mmol/l) the features of liver steatosis were found in 50% of cases. Both in the groups of patients with hypercholesterolaemia and with mixed HLP, the patients with liver steatosis had significantly higher body mass index. In the group of patients with hypercholesterolaemia (without and with the features of liver steatosis) no differences were found in the concentrations of total cholesterol (TCH), triglycerides (TG) and high density lipoprotein fraction (HDL). In the patients with mixed HLP and the features of liver steatosis in USG examination, the mean serum triglyceride concentration was 6.2 mmol/l and was almost twice higher than that in the group without steatosis. The serum HDL concentration in the patients with mixed HLP was 39.2 mg% and was significantly lower than that in the patients with the same type of lipid concentration disturbances but without liver steatosis (48.2 mg%). The total serum cholesterol concentration was not differing significantly between the patients with liver steatosis and those without this pathological condition. PMID- 7483619 TI - [Evaluation of the effectiveness of the preparation Panzytrat 20,000 U in the substitute treatment of chronic pancreatitis]. AB - Panzytrat 20,000 U produced by Knoll is a modern galenic form of active pancreatic enzymes in the form of microgranules with uniform enzymatic composition. The studies of the preparation effectiveness were carried out in 88 patients with diagnosed exocrine failure in the course of chronic pancreatitis. The patients were taking Panzytrat 20,000 for 4 weeks in doses 3 x 1 capsule with main meals. Control examinations were performed two and four weeks after starting the treatment, assessing the patients' condition on the basis of history taking and medical examination; taking into account weight gain, duration and intensity of pain and dyspeptic symptoms, frequency and character of defeacations and possible undesirable effects. In a group of 37 patients, BNT-PABA test was also performed before the treatment and after one capsule of Panzytrat. The treatment with Panzytrat 20,000 caused a statistically significant reduction of the number of defecations, significant reduction of pain and dyspeptic symptoms. A statistically significant increase of PABA recovery in urine was also obtained after one capsule of the preparation. Practically, no unfavourable effects of the treatment were observed. The studies carried out point to the high effectiveness of Panzytrat in dose 3 x 20,000. U daily in the treatment of exocrine pancreatic failure in the course of chronic pancreatitis. PMID- 7483618 TI - [Fatty liver assessed by histologic examination in patients with hyperlipoproteinemia]. AB - Out of 665 patients after liver biopsy performed during 14 years of the work of the Clinic, 35 cases of patients with non-alcoholic liver steatosis and hyperlipoproteinaemia were selected and analysed retrospectively. The cases of patients with the presence of HBV or HCV infection markers were excluded. The histological material was divided according to the intensity of steatosis expressed as the per cent of hepatocytes with the features of fatty degeneration and also according to the following classification: I--steatosis, II--steatosis with hepatitis, III--steatosis with portal fibrosis, IV--steatotic cirrhosis. In the patients in whom the per cent of hepatocytes with fatty degeneration in biopsy examination exceeded 60%, the mean serum triglyceride concentration was 5.53 mmol/l and was over twice higher than that in the group with steatosis not exceeding 30% of hepatocytes. Similarly, in the patients with steatosis and accompanying hepatitis (steatohepatitis), the mean triglyceride concentration was 5.28 mmol/l and was over twice higher than that in the patients with steatosis with accompanying portal fibrosis. The patients with steatohepatitis had significantly lower HDL concentrations than those with the remaining types of histological changes. PMID- 7483620 TI - [Clinical treatment results of treating androgenization symptoms in women with preparation Diane-35]. AB - 26 women with androgenization: hirsutism, acne, seborrhea, aged 18-30 were treated with Diane 35 during at least 12 cycles. In investigated group of patients acne was in remission in 82.4% after 3 cycles, seborrhea was reduced in 85.5% after 3 cycles and in 96.2% after 6 cycles. Hirsutism was reduced only in 19.2% women after 12 cycles. Diane 35 was well tolerated and effective in contraception all investigated women. PMID- 7483621 TI - [Clinical observations using the new contraceptive preparation containing gestagen]. AB - 31 women aged 26-35 were observed during hormonal contraception with Femovan (in Poland registered as Femoden). The aim of the study was an analysis of effectiveness, influence on menstrual cycles and adverse effects in Femovan users. The conclusions of the study indicated that Femovan is very effective in contraception (Pearl index = 0). It provides good cycle control. It reduced in 50% women nausea and breast tenderness which occurred before starting of contraception. There were not body gain and elevated blood pressure in any women. Acne was reduced in 40% women after 3 months of using. PMID- 7483622 TI - [Results of intra-arterial administration of PGE1 prostaglandin]. AB - On the basis of own clinical material the results are presented of intra-arterial administration of PGE1 prostaglandin (Prostavasin) to 18 patients with chronic lower limb ischaemia in II, III and IV stage of the disease according to Fontaine. In the group of 15 patients (four treated as outpatients) Prostavasin was administered in multiple infusions of 20 micrograms of the drug, according to the schema recommended by the producer. Three very good, six good and three poor results were observed. The best results were seen in young patients with persisting pulse over the popliteal artery. The presented method of treatment was assessed as safe, with the possibility of application, in selected cases, on outpatient basis. PMID- 7483624 TI - [Evaluation of the effectiveness of cefazolin++ (Kefzol) in preventing infections in patients after abdominal surgery]. AB - The purpose of the work as an assessment of the effectiveness of cefazolin (Kefzol) in preventing infections in patients after abdominal surgery. The prospective studies included 200 patients, out of whom in 100 patients in the studied group Kefzol was administered prior to and for 24 hours after operation, while in 100 patients in the control group no prevention was used. The percent of infections in the studied group was lower in comparison to the control group (9% vs 25%, p < 0.01). A statistically significant reduction was obtained of wound infection per cent from 19% in the control group to 7% in the studied group, including a reduction from 14.3% to 3.1% after operations on the gall bladder and bile ducts, and from 18.2% to 11.1% after gastroduodenal operations. The per cent of wound infections after appendectomy was 5.9% in the patients from the studied group receiving prophylactically Kefzol with metronidazole, and 20.6% in the patients from the control group. Bacteriological investigations, demonstrated that 87.5% bacterial strains were susceptible to cefazolin. In the authors' opinion cefazolin (Kefzol) is effective in the prevention of purulent complications, and the reduction of the per cent of infections causes lowering of treatment cost. PMID- 7483625 TI - [Assessment of gentamicin dosage in patients before planned surgical operation]. AB - The study included 18 patients, whom before planned surgical operation administered preventively gentamicin in the dose 3 x 80 mg/24 h i.m. After 24 hours the venous blood was taken before administration of the next dose of the drug and the concentration of gentamicin was determined by FPIA method. The subtherapeutic level of the drug was found in 15 patients (83%), therapeutic level - in 2 patients (11%) and toxic level in 1 patient (6%). It seems, that in patients after surgical operation, in whom after the prophylactic administration the treatment of gentamicin is continued, it is necessary the monitoring of the concentration of this antibiotic. PMID- 7483623 TI - [Evaluation of the usefulness and technique of intraoperative large bowel lavage]. AB - The technique is presented of intraoperative large bowel lavage, making possible cleansing of the bowel in patients in whom this was impossible to perform in the preoperative period. This method in some patients enables abandoning of diverting fistula creation and thus avoiding of multistage treatment. PMID- 7483626 TI - [Perception of family by children and youth with chronic renal failure]. AB - The perception of family by children and youth with chronic renal failure treated conservatively and with haemodialyses was studied. Intrafamilial relations were subjected to analysis. It was found that with the limitations due to the disease, family is the main environment where the psychophysical needs of the child are fulfilled and it is of decisive value for the quality of life of patients. In view of the importance ascribed to it, it may play a significant role in the accommodation of the patient to the disease, influencing the effectiveness of long-term treatment. PMID- 7483628 TI - [Diagnostic difficulties and clinical manifestations of mitral valve leaflet prolapse syndrome in children]. AB - The syndrome of mitral valve leaflet prolapse into the left atrium is a frequently observed during developmental period circulatory system anomaly. In the paper, two cases of the syndrome are presented differing in symptomatology, and diagnostic difficulties connected with that are discussed. PMID- 7483627 TI - [The role of nutrition in the development of circulatory system diseases]. AB - In the paper the authors synthetically discuss the aetiology and pathogenesis and point to the possibilities of prevention of one of the most important groups of diseases developing as the result of faulty nutrition--circulatory system diseases. In the first part the epidemiology is discussed of the system's diseases--ischaemic heart disease, hypertension, cerebrovascular diseases- stressing the extent of the health problem caused by them--over 50% of deaths each year are due to these diseases. Then, the main cause is discussed of the development of cardiovascular system diseases, that is atherosclerosis. The risk factors are characterized in which nutrition plays an essential role - blood level of cholesterol, its content in low (LDL) and high (HDL) density lipoproteins, triglyceride level, content of saturated fatty acids in diet. Nutritional preventive factors are briefly described--mono- and polyunsaturated fatty acids, vitamin E, cellulose. The beneficial effect of increased fish consumption is stressed. The last part deals with arterial hypertension as the disease of the vascular system and as the risk factor of ischaemic heart disease. The attention is paid to the necessity of changes in nutrition mode--increase of the supply of mono- and polyunsaturated fatty acids with predominance of the former, decrease of the supply of saturated fatty acids, salt, ensuring of adequate supply of antioxidant vitamins and cellulose. In the paper it is stressed that the modern state of knowledge, used in practice may significantly decrease in Poland, through nutrition, the epidemic of circulatory system diseases similarly as it has been done in other countries. PMID- 7483629 TI - [A case of Lesniowski-Crohn disease preceded by Yersinia pseudotuberculosis infection]. AB - In etiopathogenesis of Lesniowski-Crohn disease we consider environmental genetical, immunological and physical factors. Among the bacteria that are of importance in a disease process there are: Jarsinia, tubercle bacillus and para- tubercle bacillus. We are to state that the pro.... of the bacteria per se doesn't influence the initiation of a disease. The initiation is caused by a change of the immune response. The introduced case shows the occurrence of Lesniowski-Crohn disease proceeded by Jarsinia pseudotuberculosis. Thai sequence of occurrence doesn't prove the dependence of an effect upon the cause. What is more-there are many controversial opinions in literature considering Lesniowski Crohn disease. That is why we would like the introduced case to be a contribution to a discussion on the subject of: Possibility of a distant initiation of a disease by some bacterial infection types. PMID- 7483630 TI - [Extensive, bleeding skin cancer on the back in a postoperative scar]. AB - A case is described of extensive, bleeding plano-epithelial carcinoma of back skin in burn scar in a 62-year-old man. The lesion was extensively excised with partial resection of Th 12-L3 spinous processes with covering of the loss with a graft. During 15-month follow up no manifestations of disease recurrence were seen. PMID- 7483631 TI - [Multiple, giant retroperitoneal space lipomas]. AB - A case is presented of multiple retroperitoneal lipoma in a 59-year-old woman, displacing the left kidney to the right side. The dimensions of four tumours exceeded significantly the expected size determined in USG examination. The authors believe that in view of the size of the tumours, their multiplicity, and histological pleomorphism the patient requires further careful observation despite radical excision of the lesion. PMID- 7483632 TI - [A case of large bowel injury with a safety belt in a pregnant woman]. AB - The authors present the case of a 24-year-old pregnant woman (9th week of pregnancy) who, due to a traffic accident, suffered transverse colon rupture through a sudden compression of the intestine with a "traditional" safety belt. The authors analyse the mechanism of injury, method of treatment, and on the basis of literature data discuss various body injuries developing as a result of action of safety belts. PMID- 7483633 TI - [After vaccination for tick-borne encephalitis must onset of neurologic disorders be expected?]. AB - In Austria at present more than 5 million people are vaccinated against tick borne encephalitis (TBE), some of whom have received up to 8 vaccinations. Since introduction of the mass vaccination programme an estimated 18 million TBE vaccinations have been given in this country. Hence, the change coincidental occurrence of various illnesses which might be interpreted as possible vaccination complications is inevitable. However, the appearance of true vaccination complications in addition to chance concurrent disease cannot be ruled out a priori. In any given case it is very difficult, indeed virtually impossible, to differentiate between these two possibilities since there is no available test for confirmation or exclusion of vaccine complications. Mainly disturbances of the nervous system have been ascribed to preceding TBE vaccination. From the incidence of such diseases we calculated how often their occurrence should inevitably be expected during a two-month observation period after vaccination; only significantly higher figures can be accepted as representing vaccination complications. Very large controlled field studies would be required to provide unequivocal results. This enormous expense is barely justifiable and the required money could be better spent on other projects in the public health system. All studies to date and theoretical considerations show that the potential risk of TBE vaccination is extremely low. Experts of the Paul Ehrlich Institute (Germany) came to the conclusion that one certain case of neuritis arises after 1 million TBE vaccinations. Virological investigations show that virologically unclarified (and, hence, vaccine-induced) neurological disturbances do not occur more frequently in TBE vaccines than in unvaccinated persons.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7483635 TI - [Dermatomyositis: successful therapy with high dosage intravenous 7S immunoglobulins]. AB - This report describes the successful administration of high dose, intravenous 7S immunoglobulins (IG) to an 11 year-old girl with corticosteroid-resistant dermatomyositis. Clinical symptoms improved immediately and the pathological laboratory findings normalized within 10 days of commencement of immunoglobulin therapy. After three cycles of IG the patient showed complete remission of the muscular and dermatological findings. On follow-up examination after one year the patient remained symptom-free in the absence of any therapy and was leading a normally active life. The efficacy of IG therapy in this case is reviewed in the light of the literature. PMID- 7483636 TI - [How appropriate is laparoscopic cholecystectomy in acute cholecystitis?]. PMID- 7483634 TI - [Cocaine abuse--with special reference to cerebrovascular complications]. AB - The role of drug abuse as a risk factor for cerebrovascular events has been underestimated, particularly in patients with juvenile stroke. The drug most often associated with acute cerebrovascular events is cocaine. After cocaine hydrochloride abuse intracerebral hemorrhages or subarachnoid hemorrhages- predominantly caused by ruptured aneurysms or arteriovenous malformations--are by far the most frequently observed cerebrovascular complications, whereas on abuse with the alkaloidal form ("crack") intracranial hemorrhages and ischemic strokes are encountered with equal frequency. In most cases, the time interval between drug abuse and the cerebrovascular event is less than 3 hours. Several pathophysiological mechanisms are discussed as serving as triggers for the cerebrovascular event, either alone or in combination. No specific antidote to cocaine is known. Nevertheless drug screening (urine analysis) should be performed immediately to allow optimal management of patients with drug associated acute cerebrovascular events, especially in cases with juvenile stroke. PMID- 7483638 TI - [Departure of pharmacognosy from the Josephinum institute]. PMID- 7483637 TI - [Preoperative diagnosis of malignancy of ovarian space-occupying lesions with special reference to new, laparoscopic surgical techniques]. PMID- 7483639 TI - [1995 consensus guidelines for cholesterol. Society of Physicians in Vienna]. PMID- 7483640 TI - The clinical morphology of human atherosclerotic lesions. Lessons from the PDAY Study. Pathobiological Determinants of Atherosclerosis in Youth. AB - The PDAY study addresses the role traditional risk factors play in the development and progression of arterial lesions in youth (15 to 34 years of age). This autopsy based study indicates that in the aorta and in the right coronary artery, atherosclerosis is positively associated with cholesterol (LDL+VLDL), glucose intolerance, smoking, hypertension, and obesity and negatively associated with HDL-cholesterol. When a high cholesterol group (> 240 mg/dl) is compared to a low cholesterol group (< 200 mg/dl) early fatty sudanophilic lesions decrease in the aorta from 35% to 29% (p < 0.03) surface area, while raised atherosclerotic lesions are lowered (from 5%) in the high cholesterol group (to 3%) in the low cholesterol group; p < 0.02). Clearly, risk factors particularly cholesterol levels, directly affect the development of atherosclerosis in the coronary arteries and the aorta of young people. The data presented suggest that risk factor modification would be beneficial in young people as from 15 years of age. PMID- 7483642 TI - [Dietary treatment of hypercholesterolemia]. AB - The basis of successful treatment of the various forms of hypercholesterolemia is a balanced diet. Initially this includes a general change in lifestyle with a new approach to nutrition, an increase in physical activity and nicotine abstinence. For the individual this implies a diet containing less fat but more unsaturated fatty acids, less salt, less cholesterol, more proteins, more dietary fiber and more complex carbohydrates. Hyperlipidemia triggered by incorrect nutrition or polygenous hypocholesterolemia can usually be successfully treated by means of a fat-modified diet alone. Hypercholesterolemia due to apolipoprotein E polymorphism responds equally well to dietary change. In some cases such dietary modifications can result in a 25% decrease of LDL-cholesterol. PMID- 7483641 TI - [Drug therapy of hypercholesterolemia]. AB - The therapeutic aim in patients with LDL-hypercholesterolemia and coronary heart disease is to reduce LDL-cholesterol levels to around 100 mg/dl. Since such levels are hardly ever reached solely by diet modifications, inevitably the targeted use of cholesterol-lowering drugs is required. Drug therapy of LDL hypercholesterolemia nowadays includes the following: anion exchange resins, HMG CoA-reductase-inhibitors, fibrates, nicotinic acid and its derivatives, beta sitosterol and probucol. If cholesterol levels are not sufficiently lowered in response to monotherapy with one of these drugs combined drug therapy should be attempted. Should this likewise prove unsatisfactory then LDL apheresis can be attempted at regular intervals. This regimen usually achieves the desired LDL cholesterol level even in severe cases. PMID- 7483643 TI - Atherosclerotic lesions in humans--plaque stabilization and regression. AB - Although lowering of total- and LDL-cholesterol with different methods, like dietary restrictions mostly combined with lipid-lowering drugs (monotherapy or combined drug therapy) or changes in lifestyle (e.g., cessation from smoking) have demonstrated significant clinical benefits in a number of clinical trials, the magnitude of angiographically proven regression of atherosclerotic lesions is relatively small despite aggressive lipid-lowering regimens. A potentially important therapeutic target especially for limited indications includes the use of LDL-apheresis, where functional and later morphological regression occurs. PMID- 7483644 TI - [Fibrin sealing, a concept for early elective endoscopic therapy]. AB - The technique of fibrin adhesion is a standard haemostatic procedure which uses a two-component adhesive consisting of highly concentrated fibrinogen and thrombin, which when mixed, form a fibrin clot via the third phase of the blood coagulation cascade, and thereby induce tissue-reparative mechanisms. In a new endoscopic procedure for the treatment of bleeding ulcer, the components of the fibrin adhesive are simultaneously injected into the tissue beneath and around the lesion where they form a mechanically resilient and stable fibrin clot. This clot is firmly anchored into the tissue. In contrast to other injection methods, this "bioidentical" type of procedure can be safely repeated, allowing multiple injections to be applied to achieve increased stability and better control. Supported by a scheme of close follow-up endoscopic examinations permitting any number of subsequent therapeutic and prophylactic injections to be given for persistent bleeding stigmata, the new treatment method proves to be not only particularly suitable for preventing rebleeding but, for the first time, also enables the limited time interval after initial haemostasis to be used for genuine prophylactic therapy, hence "early elective" prophylaxis. For the patient this prophylactic form of endoscopic therapy has the advantage that emergency surgery and potential rebleeding can be avoided, in addition to which there is a clear cost-to-benefit advantage as the patients usually can be treated as outpatients or day patients, and at least with a definite reduction in hospital stay. The following report describes this new treatment concept. PMID- 7483645 TI - [Fibromyalgia]. AB - Fibromyalgia, also called (primary) fibromyalgia syndrome or fibrositis (syndrome), is a chronic soft tissue pain syndrome characterized by the presence of widespread musculosceletal aching, tender points at characteristic sites, fatigue, and poor sleep. The etiology of this common and painful condition is incompletely understood, non-restorative sleep, which has been correlated with an electroencephalic abnormality called alpha-delta sleep (intrusion of alpha rhythms in the non-REM sleep EEG), may be important. Several observations suggest that serotonin is likely to mediate both pain and non-REM sleep, and serotonin deficiency in the brain has been suggested to be of pathogenetic significance in fibromyalgia syndrome. A lower than normal threshold for pain can often be observed. The newly proposed criteria for the classification of fibromyalgia syndrome (widespread pain in combination with tender points at 11 or more of 18 specific tender point sites) are important for the differential diagnosis and to compare results in international studies. PMID- 7483646 TI - [Migraine: dihydroergotamine nasal spray--an alternative]. AB - Dihydroergotamine (DHE) formerly had to be administered intramuscularly or intravenously to cope with the migraine attack. The introduction of a new regimen, the DHE nasal spray, a new era in the treatment of the migraine attack has started. The patient is independent of his physician and may avoid the painful attack by taking the treatment already during the aura. The speed and thoroughness of the relief from pain are directly proportional to the promptness with which medication is started after the onset of an attack. The recommended dose of intranasally administered DHE is 2 mg, which means two times 0.5 mg into each nostril. The effect of DHE is mediated by its affinity to the 5-HT1D receptor. DHE furthermore reacts with other 5-HT receptors and with alpha adrenergic receptors. The side effects of DHE given by nasal route are generally mild to moderate and are primarily local ones like nose or throat problems and a bitter or abnormal taste. Nausea and vomiting rarely occur. Side effect of nasal administered DHE may not be mixed up with side effects caused by ergotamine, another ergotalkaloide with a distinct profile of effects. PMID- 7483647 TI - [Social medicine aspects of pre- and postmenopause]. AB - Due to the increasing life expectancy most women nowadays suffer from menopausal problems. These include not only menopausal symptoms, but also osteoporosis and atherosclerosis. Hormone replacement therapy therefore is a measure for keeping physiological standards. It has both preventive and therapeutic importance, for the individual as well as for the public. Menopause Management is a task for lifestyle medicine, as a combination of pharmaceutical treatment and lifestyle changes. The women and physicians are working in a menopause managing team, and prevention and treatment of diseases and disorders are not separated. PMID- 7483648 TI - [Tachycardia and hypotension attacks as the main symptoms of pheochromocytoma- case report]. AB - Pheochromocytoma is a rare disorder. It usually presents typical hypertensive crises due to katecholamine secretion. Some rare cases of pheochromocytoma have been described to secrete mostly adrenalin and cause characteristical attacks of hypotension and tachycardia. The patient presented here is one of these rare cases. In patients with similar symptoms it is important to consider the possibility of an adrenalin-secreting pheochromocytoma when establishing a diagnosis. PMID- 7483649 TI - A Muslim perspective on female circumcision. AB - Western observers are unable to understand why women would want to practice clitoridectomy, just as they are perplexed at the vocal, if mostly inarticulate, rejection by many Muslims of the Cairo conference. The battle lines which get drawn have on one side public health professionals, development organizations, and feminists and on the other side conservative and "fundamentalist" Muslims who, if they are heard at all, sound impossibly antediluvian. Many Muslims, including myself, are uncomfortable with both sides. What is needed is an alternative to this polarization. The alternative I propose is the Islamic legal discourse, which might best be described as the discursive arena in which issues of societal importance get worked out. That positive change can come about from within--using the Islamic discourse--is possible because Islamic discursive systems are broad and nuanced enough to accommodate a wide variety of medical and public health endeavors. Meaningful social change and improved public health could come about by stimulating and recovering the many Islamic sunnah (exemplary) practices which are so conducive to physical and material well-being. By dealing change through existing, and proven, traditional formats, Muslims would be able to effect valuable and meaningful change in their communities. Muslim communities should not become dependent on and indentured to Western agencies and their own nation-states to solve the problems they face, including the tragic consequences of widely practiced infibulation and clitoridectomy; instead we need to apply our own traditional practices and to support an indigenous Islamic legal discourse. PMID- 7483650 TI - Women's narratives in primary care medical encounters. AB - OBJECTIVE: This study asked how women's narratives of social and emotional problems are dealt with in primary care encounters. Our conceptual work extended perspectives from narrative analysis to focus on elements of ideology, social control, underlying structure, and features of discourse that appear marginal to medicine's technical tasks. METHOD: Based on a critical review of both quantitative and qualitative techniques in research on patient-doctor communication, we developed an interpretive method with criteria to guide sampling, transcription, interpretation, and presentation of findings. We applied the method to encounters which were selected randomly from a data base derived from a stratified random sample of 336 audiotaped encounters involving patients and primary care internists. FINDINGS: As shown by illustrative encounters, women's narratives in primary care often express emotional troubles related to gender roles. Medical discourse in these encounters tends to marginalize contextual issues that contribute to women's distress. Such discourse conveys acceptance of reproductive commitments, medical management of psychic distress, and lack of criticism regarding social problems that affect women. CONCLUSIONS: Narrative analysis clarifies the medical processing of women's emotional problems that derive in large part from the social context of medicine. Women patients and medical practitioners often deal with these issues in primary care encounters rather than in psychiatric settings. These findings suggest directions for improving the discourse of primary care encounters. PMID- 7483652 TI - Barriers to condom use and barrier method preferences among low-income African American women. AB - Low-income African-American women (N = 178) entering health clinics completed surveys assessing perceived barriers to condom use for themselves personally and for African-American women generally. Following the survey, each woman received a demonstration of five barrier contraceptive methods and then rated her preference among those methods. The women perceived relatively few personal barriers to use of the male condom but perceived significantly greater barriers for other African American women (all p < .0001). The male condom was first choice of the largest percentage of women (45%) and last choice of the smallest percentage of women (11%). The male condom was preferred for its convenience, availability, and safety, although the necessity for active cooperation by the male partner was considered a hindrance to using the method. Only 23% of women ranked the female condom as first choice and 35% ranked the female condom as last choice. Reasons for selecting the female condom included preference for a female-controlled method, safety, and protection. However, the female condom was perceived to be uncomfortable, to require the partner's acquiescence, and to interfere with sexual experience. Differences in the women's perceptions of barriers to condom use for themselves and for other African-American women are consistent with Weinstein's theory of optimistic bias. Preferences among barrier methods indicate that further research and product development are needed to develop barrier methods that are female-controlled, do not require the awareness of the male partner, and are safe, comfortable, and convenient. PMID- 7483651 TI - Persistent fatigue in well women. AB - Fatigue is the subjective report of a sustained sense of exhaustion with reduced motivation and capacity for physical and/or mental activity. Although factors associated with fatigue have been explored in specific patient populations, minimal study has been devoted to exploring both the magnitude of fatigue and associated variables among women generally thought to be well. This cross sectional, descriptive study examined the relationship of behavioral, socio demographic, and emotional factors to subjective ratings of fatigue among women in the community who perceived themselves as being fatigued but otherwise healthy. A convenience sample of 155 adult women completed the Piper Fatigue Self Report Scale (PFS), the Beck Depression Inventory (BDI), and an investigator designed questionnaire that collected behavioral and socio-demographic data. Correlation and regression analyses were used with fatigue as measured by the PFS as the outcome variable. Statistically significant relationships were noted between PFS and BDI scores as well as PFS and sleep pattern, rest quality, and perceived stress. Construction of a multiple regression model revealed an adjusted R2 of .43 with the BDI score serving as the major predictor variable for fatigue. Persistent fatigue is a consistent element in the lives of many women. As such it can significantly undermine women's quality of life. PMID- 7483653 TI - Injuries to women in the United States: an overview. AB - This review summarizes the descriptive epidemiology of injuries among women in the United States, highlighting major problems as well as needs and opportunities for intervention and research. Injury mortality rates for 1984-88 were calculated from the National Center for Health Statistics mortality data tapes. Additional injury mortality and all injury morbidity information were derived from existing literature. Studies providing gender-specific U.S. injury information during the last ten years were reviewed. Injuries are the leading causes of death for females to age 34 and are responsible for more years of potential life lost than any other cause of death. The lifetime cost of injuries to females is approximately 50 billion dollars annually. Motor vehicle related injuries, falls, and violence are the most significant injury problems for women. Although morbidity is far greater than mortality, access to information about nonfatal injuries is extremely limited. What evidence does exist points to the importance of domestic assault as a major, underrecognized source of preventable injury. Though the greater magnitude of injury among men frequently eclipses the significance of injury as a problem for women, this paper presents evidence that injury is a problem which should feature prominently in the women's health agenda for the nation. There are pressing research needs to understand the changing trends in injuries to females and to identify appropriate intervention strategies. In addition, the study points to the needs for improvement in data systems to document injury morbidity. PMID- 7483655 TI - Suppression of pulmonary P4502b and induction of hepatic, intestinal and kidney P4501a-1 and 1a-2 in the Ah-responsive and non-responsive mouse by Aroclor 1254. AB - 1. Investigations in our laboratory have demonstrated a rapid suppression of the P4502b isoform in mouse lung, concomitant with significant induction of this enzyme in liver from these same animals. The current study was designed to determine whether the suppression by polychlorinated biphenyls of pulmonary P4502b required the presence of a functional Ah receptor, and additionally to delineate the time course of the induction responses to Aroclor 1254 in the liver, kidney, and intestine of the AH-responsive and non-responsive mouse. 2. P450s were quantified by specific enzyme assay and immunoblot in liver (1a-1, 1a 2, 2b), lung (1a-1, 1a-2), kidney (1a-1, 1a-2, 2b) and small intestine (1a-1, 2b) of C57 and DBA animals at varying times (48 h-12 weeks) following a single intraperitoneal dose of Aroclor 1254 (250 mg/kg). 3. The suppression of constitutive P4502b in the lung by Aroclor was observed in both strains, but was more prominent over a longer time course in the non-responsive animals. P4502b enzyme activity was increased in the liver and intestine of both strains of mouse; however, there was a significantly greater response to Aroclor in the C57 animals. These data indicate that the AH receptor does not participate in the suppression of pulmonary P4502b, and suggests that the regulation of inducible P4502b in liver and intestine is quantitatively different between these two strains of mouse. 4. P4501a was predictably induced in all tissues examined from the C57 animal, but was largely unaffected by PCBs in the DBA strain. P4501a-2, which is also regulated by the Ah receptor, was highly induced in the liver of the responsive strain, and also increased approximately two-fold in the liver of the non-responsive animals. Kidney P4501a-2 was also modestly increased by Aroclor, only in the responsive mouse. PMID- 7483654 TI - Barriers to condom use among women attending planned parenthood clinics. AB - Assessed condom use, barriers to condom use, oral contraceptive use, partnership status and STD history in 457 15-30 year-old women attending four family planning clinics. Subjects were classified into three condom use groups: Non Users (37%); Current Users (33%); and Past Users (30%). Factor analysis revealed five barriers to condom use: Partner's Perception, Peer's Perception, Pleasure/Intimacy, Communication, and Low Perceived Need. Multivariate analyses revealed significant group differences on only two barrier factors: Pleasure/Intimacy and Low Perceived Need. Low Perceived Need accounted for 13.5% of the variance in condom use. Women with low perceived need to use condoms were more likely to use oral contraceptives. PMID- 7483656 TI - Comparative disposition and biotransformation of naphthalene in fresh- and seawater-acclimated striped bass (Morone saxatilis). AB - 1. The disposition and biotransformation of naphthalene in striped bass (Morone saxatilis) acclimated to either fresh- or seawater have been determined. 2. The 24-h total concentration factor did not significantly differ between the freshwater (283.7 +/- 61.5 [+/- SD]) and seawater (245.5 +/- 44.4)-acclimated bass. At both salinities, the viscera/gonad and liver accumulated large concentrations of residues, whereas the skin retained the largest total residue mass. 3. During the 24-h depuration period, the freshwater-acclimated bass depurated 45% of retained residue and the seawater-acclimated bass depurated 85% of retained residue. 4. Naphthalene was primarily excreted unchanged (> 99% in freshwater-acclimated bass and > 88% in seawater-acclimated bass), but small amounts of 1,2-dihydro-1,2-dihydroxynaphthalene and 1- and/or 2-naphthylsulphate were formed. PMID- 7483658 TI - Effects of bioflavonoids on hepatic P450 activities. AB - 1. The effects of tangeretin, green tea flavonoids, and other flavonoids on 7 ethoxyresorufin-O-deethylase (EROD; 450 1A), 7-pentoxyresorufin-O-dealkylase (PROD; P450 2B), p-nitrophenol hydroxylase (PNPH, P450 2E1), and erythromycin-N demethylase (ERDM; P450 3A) were examined in induced rat liver microsomes. EROD, PNPH, ERDM, and nifedipine oxidase (NIFO; P450 3A4) were examined in human liver microsomes. 2. All flavonoids tested inhibited EROD activity at higher concentrations in liver microsomes. Flavone and tangeretin were potent inhibitors of EROD, with IC50's of 0.7 and 0.8 microM respectively in rat liver microsomes and 0.15 and 16 microM respectively in human liver microsomes. The green tea flavonoid (-)-epicatechin-3-gallate (ECG) was the most potent inhibitor of EROD in human liver microsomes (IC50 = 75 microM). The effect of the green tea flavonoids on EROD was complex; in addition to inhibition at high concentrations of flavonoid, moderate activation was seen at lower concentrations. 3. 450 2B-, 2E1- and 3A-dependent activities in rat and human liver microsomes were only moderately inhibited by any of the flavonoids tested, and, in general, ECG was the most potent inhibitor for these activities with IC50's ranging from 75 to 300 microM. 4. Tangeretin inhibited EROD activity (P450 1A2) in human liver microsomes in a competitive manner with a Ki = 68 nM. Tangeretin inhibited NIFO activity (P450 3A4) in human liver microsomes in an uncompetitive manner with Ki = 72 microM. PMID- 7483659 TI - Molecular modelling of members of the P4502A subfamily: application to studies of enzyme specificity. AB - 1. Using the recently published crystal structure of a bacterial P450, namely 102 (also termed P450bm3), as a template molecular models of mammalian 2A1, 2A4, 2A5 and 2A6 were constructed. 2. Substrate interaction studies demonstrated that in keeping with known catalytic activities the putative binding sites of mouse hepatic P4502A4 and 2A5 oriented testosterone for 15 alpha-hydroxylation and coumarin for 7-hydroxylation respectively. 3. Substrate interaction studies with the putative binding site of human liver P4502A6 demonstrated that coumarin was oriented for 7-hydroxylation. However, in keeping with previous site-directed mutagenesis studies with P4502A4 and 2A5, changing a single phenylalanine residue to leucine in 2A6 gave rise to a mutant enzyme, which could bind testosterone as a substrate for 15 alpha-hydroxylation rather than coumarin. 4. Substrate interaction studies with the putative binding site of rat hepatic P4502A1 suggested that this isoenzyme would hydroxylate coumarin at the 3- rather than at the 7-position. 5. The results of these molecular modelling studies demonstrate that apparently minor modifications to P4502A subfamily amino acid sequences can result in major alterations in enzyme specificity. 6. Molecular modelling is thus a useful technique that can aid in elucidating substrate specificities of P450 isoenzymes and species differences in xenobiotic metabolism. The technique can also be utilized to complement site-directed mutagenesis studies in order to identify critical structural features of P450s and other enzymes. PMID- 7483657 TI - Metabolism of theophylline and its inhibition by fluoroquinolones in rat hepatic microsomes. AB - 1. The effects of beta-naphthoflavone, dexamethasone, phenobarbitone and isosafrole on the metabolism of theophylline by rat liver microsomes have been studied. Only beta-naphthoflavone, a known P4501A inducer, increased the rate of 1-methylxanthine formation (3-fold), whereas all the inducers studied increased the rate of 1,3-dimethyluric acid production (2.5-3-fold). 2. To study the effects of a range of fluoroquinolones on theophylline metabolism, beta naphthoflavone-induced microsomes were used, as the ratio for metabolite production rates was similar to that of untreated microsomes (4:1,3-dimethyluric acid: 1-methylxanthine at 2 mM theophylline). High concentrations of fluoroquinolones (0.5-1.5 mM) were required to affect microsomal theophylline metabolism. 1-Methylxanthine was more sensitive to fluoroquinolone inhibition by enoxacin, ciprofloxacin, norfloxacin and pipemidic acid than 1,3-dimethyluric acid; CP67015, had a significant effect on 1,3-dimethyluric acid production only; binfloxacin had no effect on either pathway. 3. Ethoxycoumarin, a rapidly metabolized substrate, was also investigated as a surrogate for theophylline in in vitro experiments. Fluoroquinolone inhibition of ethoxycoumarin O-de ethylation in beta-naphthoflavone-induced microsomes was quantitatively greater but qualitatively similar to theophylline metabolism (IC50s 440-870 microM at 2 microM 7-ethoxycoumarin). 4. These data are comparable with previous rat experiments in vivo, indicating that enoxacin, ciprofloxacin and norfloxacin have similar intrinsic activity in the inhibition of theophylline metabolism. PMID- 7483660 TI - In vitro microsomal metabolism of hydrazine. AB - 1. It has been demonstrated that hydrazine is metabolized by rat liver enzymes located in the microsomal fraction. This metabolism was reduced in the absence of oxygen or NADPH and was increased by NADH in the presence of NADPH. 2. Microsomal enzyme inhibitors, piperonyl butoxide and metyrapone, significantly inhibited hydrazine metabolism but glutathione had no affect and was not depleted. 3. In addition to P450, flavin monooxygenase may also be involved in catalysing the microsomal metabolism of hydrazine. 4. Liver microsomes prepared from either beta naphthoflavone, acetone or the isoniazid-pretreated rat did not show a significant increase in hydrazine metabolism compared with microsomes from the control rat. However, although phenobarbitone pretreatment increased overall microsomal hydrazine metabolism this was not increased relative to P450 content. 5. Hydrazine metabolism was 20-70% lower in human microsomes prepared from three individuals compared with the control rat. 6. Hydrazine is also metabolized by rat liver mitochondria but the monoamine oxidase inhibitors clorgyline and pargyline do not significantly decrease this. PMID- 7483661 TI - Characterization of benzazepine UDP-glucuronosyl-transferases in laboratory animals and man. AB - 1. The O-glucuronidation of two dopamine D1 receptor antagonists, Odapipam and Berupipam, were studied in hepatic microsomal fractions from mouse, rat, rabbit, dog, pig, and man using 14C-UDP-glucuronic acid. 2. The influence of pH, detergent, gender, drug-metabolizing enzyme inducers, and age were examined. Detergents like the zwitterionic CHAPS and non-ionic Tween 20, Triton X-100, and Brij 35 stimulated the glucuronidation rate by up to 600% of native activity with the latter being most effective. Both apparent Km and Vmax increased following detergent treatment in rat hepatic microsomes. Less marked activation of UDP glucuronosyltransferase activity was observed with Brij 35 in mouse, rabbit, dog, and pig compared with rat. In contrast, human hepatic microsomes were not stimulated by detergent treatment. 3. Marked species-dependent UDP glucuronosyltransferase activity were observed for the two compounds. In general, Odapipam exhibited higher Vmax and Km compared with Berupipam with the exception of rabbit where the reverse was true. Similar kinetic parameters were, however, observed in human hepatic microsomes. Highest glucuronidation rate (in general) was observed in mouse followed by dog, pig, rabbit, man, and rat. 4. UGT activity in human livers showed up to a seven-fold variation. Conjugation of each compound were highly correlated (r = 0.92; n = 20) suggesting that identical isoform(s) were involved in this reaction. A significant age-related decrease in UDP glucuronosyltranferase activity was observed, which partly could be explained by a preponderance in elderly female donor liver samples. PMID- 7483663 TI - Cation radicals of organosulphur compounds. AB - 1. Biological and biomimetic oxidations of thioethers are reviewed. 2. gamma Radiolysis, pulse radiolysis, photochemical, chemical, and electrochemical methods for generating sulphur cation radicals are discussed and exemplified. 3. The major reactions of sulphur cation radicals: nucleophilic attack, electron transfer, decarboxylation, reaction with O2, C-S, C-C, and alpha-C-M bond cleavages, sulphur abstraction, and rearrangements are presented. PMID- 7483662 TI - Enantioselective metabolism and pharmacokinetics of Casodex in the male rat. AB - 1. Casodex, a non-steroidal antiandrogen, is a racemic mixture of R-Casodex, the pharmacologically active (-)-enantiomer, and S-Casodex, the inactive (+) enantiomer. Single oral doses of pseudo-racemic 14C-Casodex (10 mg/kg), prepared from mixtures of either 14C-labelled R-Casodex and unlabelled S-Casodex, or 14C-S Casodex and unlabelled R-Casodex, were administered to the intact and bile duct cannulated male rat. 2. Neither enantiomer underwent stereochemical inversion, but the pharmacokinetics of Casodex showed marked enantioselectivity. 3. After dosing R-labelled Casodex, plasma concentrations of R-Casodex increased slowly to reach a peak of 3.50 +/- 0.05 micrograms/ml (mean +/- SEM) at 12 h and, thereafter, declined monoexponentially with an elimination half-life of 24 h. Plasma concentrations of S-Casodex rose rapidly to reach a much lower peak of 0.97 +/- 0.06 microgram/ml at 3 h and, thereafter, declined rapidly, although there were insufficient data to determine the half-life. R-Casodex had a much higher AUC0-24 (66 micrograms.h/ml) than S-Casodex (12 micrograms.h/ml). Plasma drug concentrations measured using an achiral assay were in very good agreement with the sum of the enantiomer concentrations throughout the profile. R-Casodex comprised 94% of the total plasma radioactivity at 12 h, decreasing to 75% at 120 h. 4. Plasma concentration data generated after administration of S-Casodex were similar to those observed after dosing R-labelled Casodex. S-Casodex comprised about 74% of the total plasma radioactivity at 6 h and only 41% at 24 h. 5. The urine of intact animals contained 36 +/- 2 and 48 +/- 3% of the dose respectively up to 48 and 120 h after dosing with R-labelled Casodex, and 33 +/- 4 and 34 +/- 4% respectively after dosing with S-labelled Casodex. The urine and bile of the cannulated rat contained 43 +/- 2 and 21 +/- 2% of the dose respectively up to 48 h after dosing with R-labelled Casodex and 37 (n = 2) and 50% respectively after dosing with S-labelled Casodex. 6. After dosing with R- or S-labelled Casodex, the urinary radioactivity consisted of the carboxylic acid metabolite formed by hydrolytic cleavage at the amide, whereas biliary radioactivity consisted of hydroxy-Casodex and Casodex, mainly conjugated with glucuronic acid. The clearance of R-Casodex by each of these pathways of metabolism was less than that of S-Casodex, with direct glucuronidation and hydroxylation showing greater enantioselectivity than hydrolysis. PMID- 7483664 TI - Fragmentation reactions of aromatic cation radicals: a tool for the detection of electron transfer mechanisms in biomimetic and enzymatic oxidations. AB - 1. Mechanistic criteria, based on the side-chain fragmentation reactions of aromatic cation radicals, involving the cleavage of a beta bond (i.e. C-H, C-Si and C-S) have been developed for the detection of electron transfer mechanisms in oxidative processes of alkylbenzenes and aromatic sulphides. 2. For benzylic oxidations, the distinction between electron transfer (ET) and hydrogen atom transfer mechanism (HAT) has been based: (a) on studies of intramolecular selectivity, which, with appropriate substrates (5-Z-1,2,3,-trimethylbenzenes and 4-Z-1,2-dimethylbenzenes, where Z = OMe, alkyl), turns out to be much higher in ET than in HAT processes; and (b) on products studies concerning the reactions of bicumyl and benzyltrimethylsilanes since in these systems, the nature of products can be significantly different for ET and HAT mechanisms. 3. These criteria have been applied to the reactions of alkylbenzenes with an NO3 radical (shown to be an ET process) as well as to the microsomal and biomimetic (by iron porphyrins in the presence of PhIO) side-chain oxidation of the same compounds, where the mechanistic probes have suggested a HAT mechanism, with the exception of the biomimetic oxidation of 4-methoxybenzyltrimethylsilane in CH2Cl2-H2O-MeOH, which probably occurs by an ET mechanism. 4. For the enzymatic and biomimetic oxidation of aromatic sulphides an oxygen transfer is suggested, since, with cumyl phenyl sulphide and 4-methoxybenzyl phenyl sulphide, these reactions lead exclusively to the corresponding sulphoxides and sulphones, whereas the same substrates, in genuine ET reactions, form cation radicals which undergo C-H and C-S bond cleavage. 5. An oxygen transfer mechanism is also likely in the biomimetic and enzymatic oxidations of sulphoxides since in these reactions 4-methoxybenzyl phenyl sulphoxide is exclusively converted to sulphone, whereas in ET reactions it forms only C-S bond cleavage products. PMID- 7483665 TI - Radical intermediates during degradation of lignin-model compounds and environmental pollutants: an electron spin resonance study. AB - 1. I discuss the following aspects of Phanerochaete chrysosporium ligninase, the enzyme that has been shown to degrade a number of lignin-model compounds, and environmentally significant polycyclic aromatic hydrocarbons and polychlorinated phenols. 2. The primary mode of oxidation involves formation of a cation radical, which undergoes either hydrolysis or carbon-carbon bond cleavage. 3. In the presence of reducing substrate such as oxalic acid, ligninase has reductive activity that has been shown to be responsible for radical-mediated degradation of halogenated chemicals. 4. Electron spin resonance technique can be used to monitor the cation radical and other radical formation during ligninase-mediated degradation of lignin-model compounds and other chemicals. PMID- 7483666 TI - Central role of radical cations in metabolic activation of polycyclic aromatic hydrocarbons. AB - 1. Development of the chemistry of polycyclic aromatic hydrocarbon (PAH) radical cations has provided evidence that these intermediates play a major role in the metabolism of PAHs by P450 and in their binding to DNA. 2. Fluoro substitution of benzo[a]pyrene (BP) represents a suitable probe for studying mechanisms of oxygen transfer in the P450-catalysed formation of quinones and phenols from BP. Formation of BP-1,6-, -3,6- and -6,12-dione from the metabolism of 6-fluoroBP (6 FBP) is mediated by the intermediate 6-FBP+. Similarly, metabolism of 1-FBP and 3 FBP by rat liver microsomes produces BP-1,6-dione and BP-3,6-dione respectively. These results demonstrate that formation of quinones and phenols occurs via an initial electron transfer from BP to P450 and subsequent transfer of oxygen from the iron-oxo complex of P450 to BP. 3. Radical cations also play a major role in the formation of DNA adducts by the potent carcinogens 7,12 dimethylbenz[a]anthracene (DMBA), BP and dibenzo[a,l]pyrene (DB[a,l]P). In the binding of BP both in vitro and in vivo, 80% of the adducts are formed by one electron oxidation, namely, 8-(BP-6-yl)guanine (BP-6-C8Gua), BP-6-N7Gua and BP-6 N7adenine (Ade), and are lost from the DNA by depurination. For DB[a,l]P, depurinating adducts formed from the radical cation, DB[a,l]P-10-C8Gua, DB[a,l]P 10-N7Gua, DB[a,l]P-10-N7Ade, and DB[a,l]P-10-N3Ade comprise 50% of the total DNA adducts. For DMBA, 99% of the adducts are depurinating adducts formed from the radical cation, 7-CH3BA-12-CH2-N7Gua and 7-CH3BA-12-CH2-N7Ade. 4. In summary, radical cations of PAHs play a major role in both the metabolism and metabolic activation leading to formation of DNA adducts that are critical in the mechanism of tumour initiation. PMID- 7483667 TI - Radical cation intermediates in N-dealkylation reactions. AB - 1. A number of mechanistic possibilities exist for P450-catalysed N-dealkylation and have been considered over the years, including C- and N-hydroxylation and sequential electron transfer (SET). With peroxidases the evidence strongly favours SET and free radicals can be detected. Any mechanism must account for lack of incorporation of label from H218O into product by P450s and the high kinetic deuterium isotope effects that are seen in N-dealkylation reactions catalysed by peroxidases but not P450s. 2. Several lines of evidence support a role for SET in P450 amine oxidations, including Hammett analysis, products of dihydropyridine oxidations, and products of mechanism-based inhibition by strained cycloalkylamines. 3. The hypothesis was considered that the P450s act via base catalysis to deprotonate the aminium radical generated by SET, since the pKa has been estimated to be approximately 9. Dihydropyridine aminium radicals have low pKa (< 4) and are generally considered to have considerable kinetic acidity. None of the haemoproteins under consideration (including the peroxidases and haemoglobin) showed high kinetic hydrogen isotope effects for the oxidation of [4-2H]- or [4-3H]-labelled 1,4-dihydropyridines. These results are consonant with the view that P450s catalyse the deprotonation of N,N-dialkylaniline aminium radicals. 4. Since low isotope effects were seen with biomimetic metalloporphyrin models as well as P450s, the deprotonation is attributed to the (FeO)2+ entity, expected to be a strong base, and not the apoprotein. Thus, the FeO moiety of peroxidases is shielded, consistent with evidence by others that SET occurs through the porphyrin edge. Both P450s and peroxidases catalysed the oxidative N demethylation of aminopyrine and N,N-dimethylaminothioanisole; however, only the peroxidases generated the stable coloured aminium radicals. 5. The rates of N demethylation of variously para-substituted N,N-dimethylanilines can be used to undertake Hammett or Marcus analysis. The former yields rho = -0.6 and the latter an apparent E1/2 of approximately 1.8 for the formal (FeO)3+ entity of P4502B1. 6. Even in the oxidation of N,N-dialkylanilines, a finite rate of N-oxidation is seen (approximately 0.1% of N-dealkylation). The simplest paradigm has N oxygenation and N-dealkylation both proceeding from a common aminium radical intermediate. PMID- 7483668 TI - On the mechanism of amine oxidations by P450. AB - 1. Experimental data previously used to support an electron/proton transfer mechanism for oxidative dealkylation of amines by P450 are critically analysed with the conclusion that the mechanistic evidence is indecisive. 2. A new mechanistic criterion recently proposed to distinguish between electron/proton transfer and hydrogen atom transfer mechanisms is discussed. It is based on isotope effect profiles determined for the deprotonation of a series of para substituted N-methyl-N-trideuteriomethyl)aniline cation radicals by pyridine and for hydrogen atom abstraction from the corresponding neutral amines by the tert butoxyl radical. These reactions model the steps proposed in the two P450 mechanisms. 3. Isotope effect profiles measured for the demethylation of substituted NN-bis(dideuteriomethyl)anilines by four different forms of P450 were found to be experimentally indistinguishable from the hydrogen atom transfer profile, and distinctly different from the cation radical deprotonation profile. This provides strong evidence that P450 oxidatively dealkylates the amines by a hydrogen atom transfer mechanism and not by an electron/proton transfer mechanism. PMID- 7483669 TI - One-electron oxidations by peroxidases. AB - 1. Peroxidases typically follow the reaction cycle: native enzyme-->compound I- >compound II-->native enzyme, in which the latter two steps involve hydrogen atom transfer from substrate to enzyme. 2. Exceptions involve (1) very facile, rapidly reacting reducing substrates that transfer an electron rather than a hydrogen atom, resulting in formation of a substrate pi-cation radical; (2) two two electron transfer steps: native enzyme-->compound I-->native enzyme; and (3) compound III and the reduced form of the enzyme containing iron(II). 3. Prostaglandin H synthase is a peroxidase with some of the properties of a P450 in that compound I can abstract the hydrogen atom from a C-H bond. 4. The so-called cyclooxygenase and peroxidase activities of prostaglandin H synthase are intimately connected and, with the above exception, both are part of a conventional peroxidase cycle. PMID- 7483670 TI - Aminium cation radical mechanism proposed for monoamine oxidase B catalysis: are there alternatives? AB - 1. The interaction of bovine liver mitochondrial monoamine oxidase B (MAO B) with a series of benzylamine analogues was investigated to provide mechanistic information relative to the proposed cation radical mechanism and to provide information on the structural requirements of the substrate binding site. 2. Steady-state kinetic analysis of MAO B with 11 ring-substituted benzylamine analogues showed substitution does not alter the reaction pathway. All amine analogues tested exhibit sizeable deuterium kinetic isotope effects. 3. Anaerobic stopped-flow kinetic studies showed (1) C-H bond cleavage is rate-limiting in enzyme-bound flavin reduction and (2) that no specially detectable flavin radicals are observed. 4. The binding affinity of para-substituted benzylamine analogues to MAO B increased as the hydrophobicity of the substituent increased. In contrast, meta-substitution of the ring showed reduced affinity with an increase in the van der Waals volume of the substituent. 5. The rate of enzyme reduction by para-substitution exhibited a strong negative dependence with the Taft (Es) steric value of the substituent. In contrast, the rate of enzyme reduction by meta-substituted benzylamines is independent of the nature of the substituent. 6. para-Substituted N,N-dimethylbenzylamine analogues are not substrates for MAO B but are competitive inhibitors of benzylamine oxidation with a weaker affinity with increasing van der Waals volume of the substituent. In contrast, meta-substituted N,N-dimethyl benzylamine analogues are weak substrates for MAO B with oxidation occurring exclusively at the benzyl carbon. 7. The consequences of these results on the possible mechanisms (aminium cation radical, H abstraction, and nucleophilic mechanism) for C-H bond cleavage proposed for MAO B are discussed. PMID- 7483671 TI - Radical cations of benzo[a]pyrene and 6-substituted derivatives: reaction with nucleophiles and DNA. AB - 1. Oxidation of benzo[a]pyrene (BP) by I2 in the presence of AgClO4 in benzene generates the BP.+ClO4-.AgI complex. This same method was used to produce radical cations from 6-FBP, 6-ClBP, 6-BrBP and 6-CH3BP. 2. Reaction of the BP, 6-FBP, 6 ClBP and 6-BrBP radical cation perchlorates with H2O produced BP 1,6-, 3,6- and 6,12- dione, whereas 6-CH3BP.+ClO4-.AgI yielded 6-CH2OHBP. 3. When BP.+ClO4-.AgI and 6-FBP.+ClO4-.AgI were reacted with NaOAc in H2O/CH3CN (9:1), 6-OAcBP was formed, in addition to the quinones. In the case of 6-ClBP.+ClO4-.AgI, a small amount of 1-OAc-6-ClBP and 3-OAc-6-ClBP was formed in addition to the diones, whereas for 6-BrBP and 6-CH3BP the reaction products were BP diones and 6-CH2OHBP respectively. 4. These results confirm the localization of charge in the BP.+ at C-6, followed by C-1 and C-3. 5. The reaction of BP with NOBF4 in CH2Cl2 produced BP.+BF4-, radical cation free of complexation with inorganic salts. 6. Reaction of BP.+BF4- with DNA produced the depurinating adducts BP-6-C8Gua, BP-6-C8dGua and BP-6-N7Gua. PMID- 7483672 TI - Porphyrin pi-cation and protein radicals in peroxidase catalysis and inhibition by anti-thyroid chemicals. AB - 1. Thyroid peroxidase (TPO) catalyses the iodination and phenolic coupling reactions in the biosynthesis of thyroid hormones. 2. The two-electron oxidation of TPO by H2O2 produces an oxoferryl porphyrin pi-cation radical compound I that isomerizes spontaneously to a form of compound I that contains an oxoferryl haem and the second oxidizing equivalent as an amino acid radical. 3. The pi-cation radical compound I is the catalytic species that effects iodide ion oxidation and the protein radical compound I is most likely the catalytic species that catalyses coupling. 4. Methimazole, a therapeutic, anti-hyperthyroid drug, is a suicide substrate for TPO and effects irreversible inactivation by TPO-mediated S oxygenation to a reactive sulphenic acid that binds covalently to the prosthetic haem. 5. Sulphamethazine and other arylamines containing electron-withdrawing substituents inhibit TPO compound I-mediated reactions by reversible, mixed-type inhibition. 6. Ethylenethiourea, a fungicide metabolite, blocks TPO-mediated iodination by reacting with the catalytic iodinating species as an alternate substrate. 7. Resorcinol and related dietary flavonoids are suicide substrates for TPO and act by covalent binding to amino acid residues, presumably those radical sites present in the compound I isomer. 8. Nitrosobenzene, a known radical-trapping agent, blocks TPO-mediated coupling but not iodination or phenolic oxidations presumably by interception of the 3,5-diiodotyrosyl radical species generated during the coupling reaction. PMID- 7483673 TI - Haemoprotein-mediated metabolism of enamines and the possible involvement of one electron oxidations. AB - 1. Microsomal metabolism of 1-benzylpiperidine (1-BP), its cis-2,6-dimethyl (cis 2,6-DMBP), 4,4-dimethyl (4,4-DMBP), and alpha, alpha-dimethyl (alpha, alpha-DMBP) analogues, and phencyclidine (PCP) has been studied to assess the involvement of P450 oxidation of the enamine tautomers of the initial endocyclic iminium metabolites. 2. The selective prevention by cyanide of the metabolite production of 1-benzyl-3-piperidone but not 1-benzyl-3-piperidinol from 1-BP is consistent with the enamine as the source of the 3-one metabolite. 3. The parent amines and particularly the independently prepared iminium species induced a pattern of metabolism-dependent irreversible inactivation of P450 benz-phetamine demethylase activity, consistent with involvement of enamine C-3 oxidation in the inactivation process. 4. Substrate activity of the endocyclic enamines and alpha aminoketones (presumably the enol-enamine tautomers) for horseradish peroxidase under conditions where simple aliphatic amines display no activity is consistent with metabolic one-electron oxidations of the enamines. PMID- 7483674 TI - Primary lymphoma of the thyroid. AB - Primary lymphoma presenting in the thyroid gland is uncommon. A review of the Yonsei University Medical Center experience between 1982 and 1994 was performed retrospectively to assess the treatment outcome and prognostic factors. There were four females and one male, and the median age was 65 years. All 5 cases presented with a neck mass. Two of them had co-existing biopsy-proved Hashimoto's thyroiditis and three cases were each in a hypothyroid state. All cases with non Hodgkin's disease were of intermediate grade. One case was in stage IE and four were in stage IIE. Three cases were treated with surgery alone and two cases with bulky inoperable stage IIB were treated with chemo-radiotherapy. Chemotherapy induced a complete response in one and a partial response in the other with minimal transient toxicity. As the questions regarding justification for extensive surgical intervention increase, combined chemo-radiotherapy can be suggested as an initial treatment even in stage I, and stage II thyroid lymphoma based on prognostic factor evaluation. PMID- 7483675 TI - Analysis of p53 tumor suppressor gene mutations and human papillomavirus infection in human bladder cancers. AB - To determine whether the dysfunction of p53 caused either by mutation of the p53 gene itself or by binding to E6 protein of oncogenic HPVs is involved in the transitional cell carcinomas (TCCs) of the bladder, we analyzed 23 TCCs of the bladder. DNA was extracted from each paraffin embedded tissue of TCCs of bladder and polymerase chain reaction (PCR)/single strand conformation polymorphism (SSCP) analysis were performed to screen mutations in p53 tumor suppressor gene, then PCR/dot blot hybridization were performed to detect infection of HPVs. We found that p53 gene mutation was found in 3 cases and oncogenic HPV infection was detected in 8 cases and thus, the overall incidence of possible p53 dysfunction was 47.8% on DNA analysis (If the results of immunohistochemistry to detect overexpression of p53 protein were included, the incidence was 60.9%). Therefore, we concluded that dysfunction of p53 plays a major role in the development of TCCs of bladder in Korean patients. PMID- 7483676 TI - The biomechanical properties of deep freezing and freeze drying bones and their biomechanical changes after in-vivo allograft. AB - This study measured the physical properties in bending of the rat femur and compression of the rat first tail vertebra subjected to deep freezing at -80 degrees C for 2 weeks, 6 weeks, 12 weeks and freeze drying. This study also measured the mechanical changes after in vivo allograft of fresh bone, deep freezing(2, 6, 12 weeks) and freeze drying. Analysis for deep freezing groups showed a mean 7.2% decrease in bending strength and 11.0% decrease in compressive strength when compared with the control group, but there was no statistical difference in the duration of deep freezing. The groups of in vivo graft after deep freezing showed 23.1% and 22.2% decrease in bending and compressive strength. There was no statistical difference in the duration of deep freezing. The freeze drying group showed a 9.7% decrease in bending strength and no significant difference in compressive strength. The group of in vivo graft after freeze drying showed a 30.1% and a 41.3% decrease in bending and compressive strength. The above results suggested that there would be some mechanical limitation in using freeze dried graft for supporting implants. PMID- 7483677 TI - Effects of iontophoretically applied naloxone, picrotoxin and strychnine on dorsal horn neuron activities treated with high frequency conditioning stimulation in cats. AB - Transcutaneous electrical nerve stimulation(TENS), acupuncture-needling, and electroacupuncture are useful non-ablative methods in medical practice for relief of pain. These procedures appear to work by causing an increased discharge in afferent nerve fibers which in turn modifies the transmission of impulses in pain pathways. It is known that the mechanism of analagesic effect via these maneuvers are variable depending on the stimulating parameters. For example, the endogenous opioid system is profoundly related to the mechanism when a peripheral nerve stimulation is applied with parameters of low frequency and high intensity. However, when stimulated with parameters of high frequency and high intensity, the reduced activity of dorsal horn neurons is only slightly reversed by a systemic administration of naloxone, a specific opiate antagonist. Thus, the present study was performed to investigate the neurotransmitter that concerns the mechanism of peripheral nerve stimulation with parameters of high frequency and high intensity. We used an iontophoretic application of antagonists of possible related neurotransmitters. The dorsal horn neuron activity which was evoked by squeezing the peripheral cutaneous receptive field, was recorded as an index of pain with a microelectrode at the lumbo-sacral spinal cord. Naloxone, picrotoxin and strychnine were applied at 200nA during a period of conditioning nerve stimulation. We observed the effects of these drugs on the change of dorsal horn neuron activities. The main results of the experiment can be summarized as follows. The spontaneous activity of dorsal horn neurons increased in the presence of glutamate and decreased with GABA. It did not change with naloxone, picrotoxin or strychnine. When naloxone was applied iontophoretically during peripheral nerve stimulation, there was no statistically significant analgesic effect compared with that of the control group. When picrotoxin was applied iontophoretically during peripheral nerve stimulation, the analgesic effect was reduced. When strychnine was applied, the analgesic effect was reduced but did not show a statistically significant difference with the control group. These results suggested that the GABAergic system may have been partially related in the analgesic action of peripheral nerve stimulation with parameters of high frequency and high intensity. PMID- 7483678 TI - Mechanism of transmission and modulation of renal pain in cats; effect of nucleus raphe magnus stimulation on renal pain. AB - Initially, when periaqueductal gray (PAG) is electrically stimulated, analgesia is induced, and this phenomenon is called stimulation-produced analgesia. Nucleus raphe magnus (NRM) as well as PAG are known to be the potent analgesic centers. NRM could modulate the nociceptive response of spinal cord neurons through spinally projecting fibers. However, as well as the above analgesic effects have been confined to the somatic pain, it was variable according to species, and the analgesic effect by NRM stimulation on the visceral pain was not yet clarified. In this study the analgesic effect by NRM stimulation on the visceral pain was examined through recording the activities of the dorsal horn neurons with renal input and renal pain, as a type of visceral pain. The renal pain was induced by ureteral occlusion or renal arterial occlusion, which in turn activated the renal mechanoreceptor or chemoreceptor. These cells had concomitant somatic input. In order to compare the effects of NRM stimulation on the renal pain with somatic pain, the somatic stimulation such as squeezing was conducted on the peripheral receptive field. The main results are summarized as follows: 1) After an electrical stimulation of NRM, spontaneous activities of dorsal horn neurons with renal input were reduced to 73.3 +/- 9.7% of the control value. 2) After an electrical stimulation of NRM, activities of dorsal horn neurons with renal input evoked by a brush, a type of non-noxious stimuli, did not change significantly. But the activities by a squeeze, a type of noxious stimuli, the activities were reduced to 63.2 +/- 7.2% of the control value. 3) After an electrical stimulation of NRM, activities of dorsal horn neurons with renal input evoked by occlusion of ureter or renal artery were reduced to 46.7 +/- 8.8% and 49.0 +/- 8.0% of the control value respectively. 4) The inhibitory effect of NRM on the dorsal horn neurons with renal input did not show any difference between renal A delta fiber and C fiber group. 5) By the electrical stimulation of NRM, the activities evoked by ureteral occlusion showed more reduction in the high threshold cell group than in the wide dynamic range cell group. These results suggest that activation of NRM can alleviate the renal pain as well as the somatic pain by modulating the dorsal horn neurons activities. PMID- 7483680 TI - Relationship between the auditory P300 and the procedural memory function in drug naive patients with Parkinson's disease. AB - We evaluated and compared procedural memory and auditory P300 event-related potential in age-matched normal controls (n = 15) and drug-naive patients with Parkinson's disease (n = 16). We used Gollin's incomplete picture test for visual procedural memory function and Tower of Hanoi puzzle for visuomotor procedural memory function. The mean latency of P300 was significantly prolonged in the Parkinsonian group than in the controls. In the neuropsychology test, the patients group revealed selective impairment of visuomotor procedural memory against preserved visual procedural memory. In the patients group, the latency of P300 was inversely correlated with performance of visuomotor procedural memory. These results suggest that prolonged auditory P300 event-related potential show the dysfunction of visuomotor procedural memory in the basal ganglia, which appears to be more selectively impaired than visual procedural memory in drug naive patients with Parkinson's disease. PMID- 7483679 TI - Prenatal fetal sex determination from maternal peripheral blood using polymerase chain reaction. AB - We have investigated the use of a nested polymerase chain reaction(PCR) assay with Y-specific sequence from the DYS 14 locus on the short arm of Y-chromosome for prenatal sex determination in the peripheral blood of 22 pregnant women who participated in the antenatal genetic diagnosis program. The sensitivity and specificity of the nested PCR using DYS 14 locus primers(Y1.5,Y1.6, and Y1.7,Y1.8) were 76.4% and 55.5%, respectively. In terms of gestational age, positive predictive values of 66.6%, 66.6%, and 80% were obtained for the first, second, and third trimester respectively. The corresponding negative predictive values were 50%, 50%, and 100% respectively. Male specific band was positive in three of the six cases of female bearing women and male specific band was negative in three of the seven cases of male bearing women during 9-16 gestational weeks showing low sensitivity. But all cases except one show the male specific band during the male fetus and all female fetuses did not show the male specific 198 base pair band during 18 approximately 40 gestational weeks. This study suggests that prenatal sex determination by PCR employing maternal peripheral blood was usually possible in late pregnancy but less reliable in early pregnancy. It seems that if we used a method separating fetal cells from maternal blood and then run PCR on these cells with DYS 14 locus primers we could make a fairly accurate fetal sex determination. PMID- 7483681 TI - Long-term follow-up of gracilis muscle transposition in children. AB - Sixteen children of uncontrollable fecal incontinence have been treated with Pickrell's gracilis muscle transposition since 1983: 12 had an imperforate anuses with multiple corrective operative procedures and 4 had traumatic destructions of anal sphincters. We report a series of 11 cases whom we followed-up over a period of 0.8 to 10.5 years (mean; 5.6 years). Seven patients were evaluated by anorectal manometry. All patients except one who had left hemipelvectomy and permanent colostomy showed nearly normal continence during the follow-up period. There was no evidence of fibrosis in the transposed muscles and the tensions of the transposed muscles were well maintained. The voluntary contractions of the transposed muscles were well maintained and efficient in all cases. The general manometric parameters did not correlate well with the functional results; however, there was a strong correlation in the S/R ratio (maximum squeeze pressure/resting pressure) with the functional results. We believe that the good functional outcome of this procedure need not only the meticulous surgical technique but also the personal motivation and the compliance with physiotherapy. In conclusion, although the gracilis muscle transposition never results in normal continence, acceptable continence can be achieved in the selected patients. PMID- 7483682 TI - Use of a 64 channel computerized cardiac mapping system in arrhythmia surgery. AB - A multipoint and computerized intraoperative mapping system has been known to be of value in improving the results of surgery for cardiac arrhythmia. It shows great potential as a new tool in the surgical intervention of the more common and lethal types of supraventricular tachyarrhythmias such as atrial flutter and atrial fibrillation. In addition, it also enhances the ability of the investigators to map and ablate the sometimes fleeting automatic atrial tachycardia. The authors developed a 64 channel computerized cardiac mapping system using a microcomputer (Macintosh IIx) and this has been used for basic research in cardiac electrophysiology as well as in arrhythmia surgery. In this system, bipolar electrograms are obtained from 64 different cardiac sites simultaneously at a sampling rate of 1 Ksample/sec and with a continuous and total data storage of up to 30 seconds. When the reference electrode is selected, delay time from the reference point is displayed on a two dimensional diagram of the heart. This system was used in one patient who underwent a surgical ablation of a ventricular tachycardia in whom we observed a ventricular activation sequence involving a variety of rhythms over several minutes. The system design permits easy expansion to a simultaneous sampling from 256 sites. This 64-channel mapping appeared to have the potential to be of great help in our understanding of cardiac arrhythmia as well as in its diagnosis and surgical treatment. PMID- 7483683 TI - Dysphagia due to mediastinal tuberculous lymphadenitis presenting as an esophageal submucosal tumor: a case report. AB - Mediastinal tuberculous lymphadenitis is rare in adults, and it is even rarer for dysphagia to be the presenting symptom of mediastinal tuberculous lymphadenitis. Mediastinal tuberculous lymphadenitis with esophageal symptoms has been presented as esophageal ulceration, mucosal or submucosal mass with ulceration, fistula or sinus formation, extrinsic compression, or displacement of the esophagus. An exaggerated form of extrinsic compression may be presented as a submucosal tumor, radiologically or endoscopically. A barium esophagography of a 34 year-old woman with painful dysphagia revealed a large submucosal tumor-like mass on the mid esophagus. The symptom was spontaneously improved over a 3-week period together with reduction of the mass size. A computed tomography of the chest disclosed an enlarged subcarinal lymph node and histologic examination of the specimen obtained by thoracoscopic biopsy brought about a diagnosis of tuberculosis. We herein report a case of mediastinal tuberculosis with unusual manifestations. PMID- 7483684 TI - [Motility disorders of the gastrointestinal tract. Highlights of the 1st European Postgraduate Course on Gastrointestinal Motility 10 to 13 June 1992 in Utrecht]. PMID- 7483685 TI - [Current problems in cholestasis. VIII. Hepatology Colloquium. Halle/Saale, 19-20 June 1992. Proceedings]. PMID- 7483686 TI - [Dipeptidyl-peptidase IV of human lymphocytes in patients with primary biliary cirrhosis and UDCA therapy]. AB - Investigations concerning changes of dipeptidyl-peptidase IV-positive (DP IV+) lymphocytes in the peripheral blood (PBL) were undertaken in 22 patients with primary biliary cirrhosis (PBC) under therapy with ursodeoxycholic acid (750 mg/day). We found that 4-8 weeks after therapy beginning the number of DP IV+ PBL was raised from a low initial suppressed level of 5-12% into a range of 18-33% corresponding to that of a healthy. Concurrently liver function parameters of the patients were improved significantly. The identity of DP IV+ and CDw26+ of peripheral T-cells could be shown by combination of the cytochemical DP IV staining with the indirect immunofluorescence of the lymphocyte marker CDw26. These results show that the UDCA-therapy in patients with PBC has a direct effect on the immune system and a positive influence on the immuno-regulative capacity. PMID- 7483687 TI - [Primary sclerosing cholangitis (PSC)--humoral immune phenomena, pathogenetic aspects and therapeutic possibilities]. AB - There are about 80% antibodies in PSC against cytoplasmatic antigens of neutrophilic granulocytes of the perinuclear type (pANCA), inconstantly there are antinuclear antibodies (ANA) too, but no antimitochondrial antibodies. The frequent association of PSC with colitis ulcerosa suggests an enterobacterial aetiopathogenesis. PSC sera show clear bands at 60-90 kD and at about 10 kD in the immunoblot with enterobacterial proteins as antigens. Antibodies against enterobacterial lipopolysaccharides and lipid A are to be found in patients with PSC corresponding to the normal collective. After long-term immunization with enterobacterial antigens PSC-like changes with circulating ANA can be induced in mice and rabbits. PSC, comparable to primary biliary cirrhosis, also reacts to treatment with ursodesoxycholic acid but it scarcely reacts to immunosuppressive therapy. At the final stage of the disease liver transplantation is indicated. In our clinic up to now 16 patients with PSC have undergone a transplantation with a one-year-survival rate of 88%. Confirmed re-manifestations of PSC in the transplant have not been diagnosed up to now. PMID- 7483688 TI - Cholestasis: pathophysiology and pathobiochemistry. AB - Clinical and morphological diagnosis of cholestasis involves a variety of different parameters, but bile fluid formation and composition of bile are usually not accessible in patients. In contrast, a pathophysiological definition of cholestasis can be based on current knowledge on the mechanisms of bile formation. Hence, causes of cholestasis may be localized in each step of the process of bile formation. 1. Inhibition of fluid formation reduces maximal secretory capacity. 2. Inhibition of the transcellular hepatobiliary transport may involve transport carriers, binding proteins, and the systems of biotransformation. 3. Mechanical obstruction or regurgitation of bile constituents due to increased permeability of the bile tract may inhibit the biliary elimination of cholephilic compounds. Consequences of the inhibition of biliary elimination are retention in the liver and the whole organism of potentially toxic compounds. Among these are endogenous compounds such as bile acids and cysteinyl-leukotrienes, but xenobiotics as well may become more toxic in cholestasis. The composition of bile and portal venous blood is altered. In experimental animals, changes in secretory function can already be observed before clinically used indicator enzymes of cholestasis increase. This functional inhibition of biliary elimination can be characterized as "subclinical" cholestasis that may, nevertheless, inhibit the elimination and potentiate the toxicity of cholephilic endo- and xenobiotics. PMID- 7483689 TI - [Diagnostic porphyrinopathies in hereditary hyperbilirubinemia]. AB - Secondary porphyrinopathias were investigated in hereditary hyperbilirubinemias of the types Dubin-Johnson syndrome (DJS), Rotor's syndrome (RS), Gilbert's syndrome (GS) and compared with the findings in alcohol-induced cholestasis. The determination of urinary coproporphyrin excretion including its isomer I and III relation allows to diagnose and differentiate DJS from the other two forms of hereditary hyperbilirubinemias. An isomer I proportion of > 80% (normal < 32%) in association with a normal coproporphyrin excretion is pathognomonic for DJS. An increased coproporphyrinuria with about 70% isomer I is found in RS; a normal urinary coproporphyrin excretion combined with an isomer I increase of about 50 70% indicates the frequent GS. Alcohol-related cholestasis reveals a distinct pathologic coproporphyrinuria and elevated isomer I proportions between 37-67%. Due to the overlapping of isomer alterations in DJS-gene carriers, patients with GS and patients with exogenous toxically-induced cholestasis, additional diagnostic criteria are required to clarify the porphyrinopathia in its clinical context. PMID- 7483690 TI - [Liver cirrhosis and chronic hepatic porphyria]. AB - Chronic hepatic porphyrias (CHP) are associated with different degrees of liver damage. They range from minimal histological changes to cirrhosis and primary liver carcinomas. Forty patients with early stages of porphyrias were compared to 85 patients with clinically manifest porphyrias, Porphyria cutanea tarda. This comparison resulted in the fact that cirrhosis occurred more often in early (or latent) stages (65%) than in PCT (31%) confirmed by laparoscopy. Thus the severity of the liver disease does not depend on the duration and intensity of cutaneous symptoms or pathobiochemical syndromes but they more often depend on the long-term influence of so-called trigger factors, among them above all alcohol and iron overloading. We conclude that the proof of latent CHP has an index function for severer liver damage. PMID- 7483691 TI - [Porphyrin-induced biliary cirrhosis in protoporphyria: indications for liver transplantation]. AB - A 51-year-old patient, who had experienced photosensitivity from childhood on, fell ill with an icterus accompanied by abdominal complaints. At the age of 46 years an erythropoietic protoprophyria had been diagnosed by an excessive protoporphyrinemia. Meanwhile, in consequence of the protoporphyria's erythrohepatic progression, a biliary liver cirrhosis developed, associated with portal hypertension and marked splenomegalie. As a remission of the protoporphyria-induced hepatobiliary complication could not be achieved by conservative medical treatment, the indication for orthotopic liver transplantation was established. The operation proceeded without complications, and the postoperative course, too, revealed no problems. With primarily good bile production, bilirubin and aminotransferases soon declined to normal values. Urinary coproporphyrin excretion was found to be normal after transplantation, only a domination of isomer I still peristed. Furthermore, a distinct elevation of protoprophyrin continued to exist both in erythrocytes and plasma as expression of the immanent enzyme deficiency typical for the genodermatosis protoporphyria. In the liver explant a micronodular transformation of parenchyma was found besides a deposition of reddish-brown protoporphyrin pigment in hepatocellular cytoplasma and Kupffer's cells, in addition to pigment casts in the canaliculi and within single portal bile ducts.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7483692 TI - [With primary liver carcinoma-associated cholestasis]. AB - Icterus is among the symptoms which occur late in carcinoma of the liver. Icterus which occurs through the obstruction of the bile-ducts may be an early symptom in favourable cases. On the basis of literature and the analysis of the case material in our clinic the incidence of cholestasis, its clinical picture and treatment in carcinomas of the liver is evaluated. The differential diagnostic importance of enzymes which are typical of cholestasis--alkaline phosphatase and gamma-glutamyl transferase--and their isoenzymes are discussed. The results obtained in our clinic are compared with those in literature. PMID- 7483693 TI - [Primary liver carcinoma with invasion of the duodenum--endoscopic diagnosis]. PMID- 7483694 TI - [Occupationally-induced liver damage--vinyl chloride-induced angiosarcoma]. AB - A 45-year-old chemical worker (exposure to vinyl chloride: 500-1612 mg VC/m3 over 248 months) fell ill with a haemangiosarcoma of the liver. This is the first case observed in workers exposed to VC in East Germany. We did not succeed in making the definite histological diagnosis until after the patient's death. The first pathological findings were revealed through the increase in the gamma glutamyl transpeptidase, through sonography and computer tomography of the liver. Numerous tumor biopsies produced either no material or healthy liver tissue, which led to difficulties in therapy. The only chance of effective treatment is an early liver transplantation. PMID- 7483695 TI - [Incidence and differential diagnosis of cholestasis in AIDS--a retrospective analysis of 73 patients]. AB - A significant cholestasis constellation (alk. phosphatase and GGT each increased to twice as much as the norm) in 73 consecutive patients with AIDS was found in the course of the disease in 29 patients (= 39.7%). In most of the patients advanced immunodeficiency with an average of 2.5 AIDS-defining diseases already existed. In the predominant number of cases it was intrahepatic cholestasis with only slight icterus. Therapeutic measures such as medicamentous treatment or high caloric parenteral nutrition were causal in two thirds of the cases; in a quarter of the cases opportunistic infections, above all disseminating mycobacteriosis were found. One third of the patients had already existing liver diseases. Liver biopsy frequently proved diagnostic and should be performed more often in the case of unclear liver findings in AIDS patients. PMID- 7483696 TI - [Treatment of cholestatic liver diseases with phenobarbital and ursodeoxycholic acid]. AB - Liver transplantation is the therapy of choice in the final stage of PBC, PSC and biliary atresia. UDCA leads to remission of the symptoms and the clinic in the early stages not only of these diseases but of cholestasis of pregnancy and of cholestasis in cystic fibrosis as well. In benign intermittent cholestasis only phenobarbital but not UDCA seems to be efficacious. The efficacy of UDCA in chronic active hepatitis and alcoholic hepatitis is not yet guaranteed. PMID- 7483697 TI - [Idiopathic ductopenia of adulthood: favorable effect of ursodeoxycholic acid therapy]. AB - A 59-year-old woman presented with chronic, clinically asymptomatic cholestasis. Idiopathic adulthood ductopenia was diagnosed on the basis of clinical, laboratory and morphological findings. A retrograde cholangiographic study demonstrated intact large bile ducts with an apparent reduction of the small caliber intrahepatic ducts. The patient was treated with ursodeoxycholic acid (dosage 750 mg/day) which led to a decrease of alkaline phosphatase and Y-GT levels (before therapy: AP ca. 650 U/l; Y-GT ca. 150 U/l; after 6 month of treatment: AP ca. 350 and Y-GT ca. 30 U/l), suggesting, that the drug might be efficacious. PMID- 7483698 TI - [Surgical technique and results in benign bile duct stenoses]. AB - Surgical treatment of benign bile-duct stenosis in the state of extrahepatic cholestasis is not only associated with increased operation risk, but demands a great deal of experience from the surgeon. The results obtained during the last 3 years in 17 patients with an average age of 51 are evaluated. Different techniques corresponding to the individual findings of the bile-duct stenosis were applied for reconstruction. In two thirds of the patients long-term drainages were necessary. PMID- 7483699 TI - [Extracorporeal shockwave lithotripsy of problem bile duct calculi]. AB - ESWL is a new gentle, very effective, poor-risk technique in treatment of extrahepatic problematic bile-duct stones. This method might substitute surgical choledochotomy to a great extent. ESWL is a new therapeutic alternative to achieve nonoperative freedom of stones or, at least, to treat biliary obstruction in intrahepatic bile-duct stones, which are not treatable by endoscopic operative methods. PMID- 7483700 TI - [Laser-induced intracorporeal lithotripsy of bile duct calculi]. AB - About 5-10% of all bile-duct stones cannot be removed by endoscopic papillotomy and mechanic lithotripsy. Intracorporal laser lithotropsy has meanwhile become a safe technique which can be used in an endoscopic retrograde way or percutaneously transhepatically. The greatest clinical experience exists in connection with pulsed dye laser systems. More recent developments are the solid state laser (Alexandrit laser) and dye laser, which include an integrated stone tissue differentiation system. With the latter system purely radiologically controlled intracorporal lithotripsy is possible in selected cases. This paper reports clinical experience with this lithotripsy system in 17 patients. PMID- 7483701 TI - [Delayed stenosis after endoscopic sphincterotomy (EST)?]. AB - To answer the question of restenosis following endoscopic sphincterotomy (EST) different and sophisticated methods were performed. Using the Erlangen type papillotome we developed a practicable and standardised method for measurement of the size of the EST that is able to define a restenosis. Depending on the indication for EST we found different percentages of restenosis: common bile-duct stones 14%, papillary stenosis 23.1%, duodenal diverticula without bile duct stones as cholestatic entity 40%. On one hand reduction of EST size is not corresponding to clinical symptoms, on the other hand the symptoms of patients after EST with sufficient EST size have to explained in other ways such as gall bladder still existing. Due to these experiences in patients with symptoms following EST we propose a control ERC combined with mechanical measurement of the sphincterotomy size. PMID- 7483702 TI - [Late results of endoscopic sphincterotomy in patients with intact gallbladder]. AB - Late complications and the recurrence rate of gall-bladder stones were registered by means of annual follow-up examinations over a maximum period of five and a half years in 91 out of 116 patients in whom endoscopic sphincterotomy (EST) with the gall-bladder in situ was performed. It turned out that only one patient had colic after EST in the group of patients with patent ductus cysticus (n = 36). It was not necessary to cholecystectomise any patients. In contrast, there were 13 cases of patients with incompetent ductus cysticus (n = 55) having a colic or cholecystitis and 5 cases where the patients had to be choletectomised after EST. Thus the evaluation of the ductus cysticus is of prognostic importance in making a decision to carry out cholecystectomy after successful EST. PMID- 7483703 TI - [More successful and cost effective--the non-transendoscopic method of TPCD]. AB - With TPCD best results are gained with large diameter endoprostheses. In this instance special duodenoscopes with a working channel of 3.2 or 4.2 mm are needed for the implantation of 10 or 12 French endoprostheses. Alternatively, using the non-transendoscopic technique large diameter endoprostheses of 14 or more French can be implanted independently from the diameter of the working channel with conventional endoscopes. We have used this method developed at our institution in 1204 patients since 1982 and compared its results with 192 patients in whom the transendoscopic technique with 7-10 French endoprostheses was administered. The success-rate (non-transendoscopic technique 94 vs transendoscopic technique 79%), early complications (5 vs 11%), method-specific mortality (0.3 vs 1%), in hospital mortality (3.6 vs 21%) and late complications (19 vs 33%) are clearly in favour of the non-transendoscopic approach. The higher rate of early complications and consecutively higher in-hospital mortality of the transendoscopic method with 7-10 French endoprostheses was mainly due to frequent episodes of early cholangitis due to insufficient biliary drainage. The higher success-rate of the non-transendoscopic approach is method specific ("Prothesenleger" guidable). A change of the endoscope is not necessary in contradiction to the transendoscopic method. Additional costs of special duodenoscopes that can be used neither for ERCP nor sphincterotomy are superfluous.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7483704 TI - [Palliative endoscopic treatment of malignant occlusive jaundice with a balloon expandable metal stent]. AB - Balloon-expandable Strecker-stents were endoscopically inserted in 13 patients (8 females, 5 men; mean age 75.2 [55-99] years) suffering from malignant obstructive jaundice. The stent implantation was difficult in one patient because of trouble to retract the balloon carrying the stent. A prompt regression of the cholestasis resulted in 11 patients. Two patients with long-distance stenoses of the common hepatic duct showed a persistent icterus due to an incomplete drainage of the stenoses. Therefore a second stent was implanted by means of transhepatic procedure in one case. Additional early complications such as cholangitis did not occur within four weeks after insertion. During the follow-up of four up to 41 weeks (mean follow-up 15.2 weeks) we observed late complications in five patients: stent migration in one patient after 10 weeks; relapsed jaundice in four patients 11 up to 41 weeks after stent implantation (the reason of jaundice was unknown for these patients were not again sent to our hospital because of their bad conditions). Six patients were alive without evidence of icterus on the date of examination. Considering data from the literature Strecker-stent implantations have been rarely performed in biliary duct stenosis till now. But the results demonstrate that an effective drainage of bile duct stenoses can be achieved in a high percentage by means of balloon-expandable metal stents. The insertion of the stent can be rather easily performed in stenoses situated in Vater's papilla and common bile duct provided that the guide wire has passed the stenosis. PMID- 7483705 TI - Stimulation of bile flow and inhibition of biliary secretion of taurocholate and leukotriene C4 in thioacetamide-induced liver fibrosis. AB - In order to study the relation of hepatic fibrogenesis to biliary elimination, female Uje:WIST rats were treated with thioacetamide (TAA). This treatment results in single cell necrosis, fibrosis, nodular parenchyma und proliferation of bile ducts. In isolated perfused livers from pretreated rats, liver hemodynamics, bile flow, and secretion of taurocholate and leukotriene C4 were determined. After TAA-pretreatment, sinusoidal efflux of glucose and pyruvate was reduced increasing lactate/pyruvate ratio threefold. Biliary secretion of [14C]taurocholate and [3H]leukotriene C4 was lowered by 47% and 35%, respectively, compared to controls. In contrast, basal bile flow was increased after TAA-treatment. The beta-adrenergic agonist isoproterenol stimulated bile flow threefold over controls. The increased stimulation was correlated with increased liver weight after TAA-treatment. We postulate that both increased bile flow and stimulation by isoproterenol are due to proliferation of bile ducts and increased ductular bile secretion partially compensating for disturbed hepatocellular secretion. PMID- 7483706 TI - [Endogenous digitalis-like factor in liver cirrhosis and cholestasis]. AB - Endogenous digitalis-like factor (EDLF), an inhibitor of membrane Na+/K(+) ATPase, is discussed to be involved in the pathogenesis of cirrhogenic portal hypertension, ascites formation and development of functional hepatorenal failure. Therefore, we investigated the serum content of this mediator in patients with liver cirrhosis Child-Pugh stage A, B, and C (n = 27) by means of enzyme immunoassay with a specific digoxin antibody. Furthermore, a correlation analysis was performed in order to find out correlations between signs of cell injury, cholestasis, synthetic cell function, ascites formation, and hepatorenal failure. Our results demonstrate that EDLF is significantly elevated in Child C cirrhosis (0.61 +/- 0.15 ng/ml) in comparison to Child A cirrhosis (0.013 +/- 0.2 ng/ml) and is also higher than in Child B cirrhosis (0.23 +/- 0.25 ng/ml). In patients without ascites EDLF (0.056 +/- 0.19 ng/ml) differs significantly from that of patients with non-complicated ascites (0.156 +/- 0.176 ng/ml) and from that of patients with therapy refractory ascites (0.66 +/- 0.17 ng/ml) or hepatorenal failure (1.56 ng/ml). There are no correlations between EDLF and renal function. Significant correlations were demonstrated for cholestasis (serum bilirubin), synthesis function (serum protein, Quick's value, cholinesterase, fibrinogen, albumin), and the degree of portasystemic encephalopathy (number connection test). We conclude that EDLF may act as a mediator in the process of progressive portal hypertension and its complications due to cirrhosis. This process of progression is caused by the inhibition of Na+/K(+)-ATPase, vasoconstriction, and endothelin secretion. PMID- 7483707 TI - [Isolation and characterization of hepatocytes from chronic cholestatic damage to rat livers after bile duct ligation]. AB - The aim of these investigations was the isolation and characterisation of hepatocytes from chronic injured rat livers after bile duct ligation. In three months old female Uje:WIST rats the distal common bile duct was double ligated. 14 days after the ligation 80% of the rats survived. The content of hepatic OH prolin was increased in relation to shamoperated control animals. The histological examination showed a marked fibrous and oedematous spreading of the portal tract as well as an extensive bile duct proliferation after 14 days. A correlation between OH-prolin content of the liver and index of fibrosis was found. 14 days after ligation the hepatocytes were isolated according to Seglen (7). The yield of hepatocytes isolated from the liver of cholestatic rats and from control animals was on average 4 x 10(8) cells with a viability of 80%. Ultrastructural the most hepatocytes showed intact organells without injury. We can conclude, that it is possible to isolate hepatocytes from chronic cholestatic rat livers with a good yield and viability as well as with an intact ultrastructure. These cells are suitable for additional metabolic investigations. PMID- 7483708 TI - [Identification of procollagen mRNA in chronic active hepatitis B]. AB - In situ hybridization is a tool for staining intracellular procollagen mRNAs with specific probes. Our study shows the amounts of procollagen mRNAs of types I and III to be increased in liver biopsies of five patients with chronic active hepatitis B as compared with five healthy controls. Parallel staining employing anti-smooth-muscle-actin antibodies was able to identify myofibroblast-like cells at the same localization where procollagen mRNAs were found. Consequently, these transformed Ito-cells might be the procollagen-producing cells. PMID- 7483709 TI - [Effect of extrahepatic cholestasis on amino acid metabolism in the animal experiment]. AB - The knowledge of pathobiochemical processes in extrahepatic cholestasis is one of the prerequisites for the application of infusion solutions adjusted to meet the requirements. Early changes of amino acid metabolism in rats after bile-duct ligation, which indicate disorders in the urea cycle are reported. For this purpose 60 male Wistar rats were laparotomized and the common bile duct was twice ligated under narcosis with pentobarbital. Exsanguination of the animals from the aorta abdominalis occurred after 3, 6, 12, 24, 48 and 72 hours. Analysis of the free amino acids in serum was performed by means of middle pressure liquid column chromatography at fully automatic analytical apparatus (Liquimat III, Kontron). RESULTS: Of 23 free amino acids investigated in serum only Ornithin and Arginin showed significant changes in their serum concentrations after ligature of the common bile duct. The increase of the serum concentration of Ornithin and the simultaneous decrease of Arginin suggests the inhibition of the urea cycle induced by resorptive jaundice. This might be due to the inhibition of the ATP dependent, mitochondrial carbamyl phosphate synthetase or to the activation of the cytoplasmatic arginase. PMID- 7483710 TI - [Rational laboratory diagnosis of cholestasis syndrome]. AB - Laboratory diagnostics efficiently applied is of decisive importance for a great deal of questions in spite of the technical and endoscopic methods which are available today in the field of examinations. By means of a screening programme consisting of ALAT, gamma-GT, ChE at a sensitivity of > 90% alterations in both the hepatobiliary system and cholestasis can be recognized with sufficient reliability. Clinical data and a defined laboratory routine programme as a second step in diagnostics results in reliable indication for distinction between obstructive and non-obstructive cholestasis which can be promoted by computer aided techniques. On the basis of such pre-selections special laboratory methods (differential diagnosis of consecutive non-obstructive cholestasis in liver diseases) or invasive methods to clarify their reason and localization diagnostics of biliary obstruction can then be applied in a well-directed manner to obtain a definite nosologic diagnosis. Effective diagnostic procedures in this three-step programme are described. PMID- 7483711 TI - [Status of ultrasound diagnosis in differential cholestasis diagnosis]. AB - Differentiating between the hepatocellular "internal" and the "surgical" or "endoscopic" cholestasis syndrome has become a domain of sonography. There is a 92 to 99% success rate, due to the possibility of diagnosing by means of sonography dilations of the gall bladder, the extrahepatic biliary tract and the intrahepatic bile ducts, as well as numerous diseases of the hepatobiliary system and of the pancreas. Congested intra- and extrahepatic bile ducts indicate the presence of a deep-seated distal obstruction, such s a prepapillary concrement, an inflammatory or malignant papillary stenosis or of a process in the area of the head of the pancreas. If the intrahepatic bile ducts alone are dilated the obstructive process has to be assumed near the portal fissure of the liver. Frequent causes are directly demonstrable tumors or metastases on the hilum or high-lying concrements. By using sonography, the level of obstruction can be determined correctly in about 90% of cases, while the cause of obstruction can be diagnosed in about 60 to 70%. In most cases intrahepatic cholestase can only be demonstrated indirectly through the absence of a bile duct dilation. Characteristic diffuse changes in the liver (e.g. fatty liver, cirrhosis, cardiac liver) or intrahepatic space claims-may be in combination with an ultrasound directed puncture-may support or confirm the diagnosis. PMID- 7483712 TI - [Influence of sonography on indications for laparoscopic cholecystectomy]. AB - Because of therapeutic consequences preoperative sonography should be performed with a check list. Nevertheless false positive and false negative findings result in cases of gall-stones in the ductus cysticus, of acute cholecystitis and thickness of the gall-bladder wall. Therefore where sonographic findings differ from the intraoperative laparoscopic situs the operating surgeon should change to conventional cholecystectomy early enough to avoid complications. PMID- 7483713 TI - [Endoscopy in cholestasis]. AB - The spectrum of causes of cholestasis ranges from microscopically recognizable damage to the parenchymal cells of the liver to obstruction of the ductus choledochus und papilla Vateri. Liver biopsy on the one hand and ERCP on the other therefore constitute the morphological methods that come closest to the pathological substrate. Obligatory sonography serves the restriction of diagnosis to a large extent. Laparoscopy is nearly always dispensable. Even in case of liver biopsy the therapeutic consequences should have an influence. Consequently, for transpapillary endoscopic methods in obstructive jaundice it is essential to remove the obstruction (papillotomy and extraction of the calculus or nasobiliary probe or intraluminal tube) if possible in one and the same session. The purely diagnostic ERC possibly causes dangerous cholangitis in mechanical icterus. Transpapillary and percutaneous transhepatic endoscopic methods are becoming more and more sophisticated and new indications are being discovered. However, the results of sonography and endoscopy in obstructive jaundice depend very much on the equipment available and experience of the investigator. He should realistically estimate what he is able to do as well as the patients' chances of recovery. In addition he should maintain contact to large departments of surgery and endoscopy. PMID- 7483714 TI - [Rare causes of cholestasis in ERC/PTC]. AB - Using case reports, attention is drawn to rare causes of cholestasis the clarification of which by ERC/PTC and by endoscopic therapy, if necessary, will clearly shown the usefulness of this method. The picture of an unusual choledocholith, the extraction of a "suture stone", the case of a Mirizzi syndrome, a cholecystocolic fistula, a cystic dilatation of the choledochus with Caroli's syndrome and status after choledochojejunostomy, the extraction of an ascarid from the papilla at biliary pancreatitis as well as the detection of a biliobronchial fistula resulting from an unsound hepaticojejunostomy are examples to underline the diagnostic usefulness of the ERC/PCT including its therapeutic capabilities and consequences in contrast to other methods, such as x-ray, ultrasound or computertomography. PMID- 7483715 TI - [Lidocaine elimination and MEGX formation after oral lidocaine administration--a practicable test for assessment of quantitative liver function]. AB - Oral load with 200 mg Lidocain was performed in 370 patients with chronic liver disease. The 120- and 240-minute Lidocain plasma concentrations as well as the 30 and 60-minute MEGX plasma concentrations, main metabolite of Lidocain, were measured by means of gas chromatography and with the commercial TDX test from the firm Abbott. No side effects caused by the load were observed and all of the patients resorbed Lidocain. Peak concentrations were found both for Lidocain and for MEGX in the 60-minute tests. Patients with liver cirrhosis of different aetiology showed significantly higher Lidocain plasma concentrations and lower MEGX values than patients with chronic non-cirrhotic liver disease. The differentiation of these two groups of patients was most successful via the determination of the 240-minute Lidocain plasma concentration. Oral load with 200 mg Lidocain has turned out to be a practicable and meaningful test for the estimation of the Cytochrom P450-dependent liver function. PMID- 7483716 TI - Caffeine elimination in cirrhotic and non-cirrhotic liver disease of different etiology. AB - Caffeine elimination was studied in 419 patients with cirrhotic and noncirrhotic liver disease of different etiology (hepatitis B virus infection n = 79; hepatitis NANB virus infection n = 74; ethanol-induced liver damage n = 143; primary biliary cirrhosis I-IV n = 63; cryptogenic liver cirrhosis n = 60) following oral administration of 366 mg caffeine. Caffeine clearance in the control group was 69 +/- 33 ml/min (age-matched healthy volunteers and patients without liver disease). Caffeine clearance in acute hepatitis B (70 +/- 60 ml/min) chronic persistent hepatitis B (81 +/- 56 ml/min), chronic aggressive hepatitis B (107 +/- 66 ml/min), posthepatitic liver cirrhosis B (84 +/- 62 ml/min), acute hepatitis NANB (94 +/- 69 ml/min), chronic persistent hepatitis NANB (122 +/- 60 ml/min), chronic aggressive hepatitis NANB (87 +/- 52 ml/min) and posthepatitic cirrhosis NANB (59 +/- 26 ml/min) is not reduced in comparison with controls. In patients with alcoholic fatty liver (127 +/- 71 ml/min, p < 0.05) caffeine clearance is enhanced, in alcoholic hepatitis (57 +/- 72 ml/min) comparable to controls and in alcoholic cirrhosis reduced (36 +/- 44 ml/min, p < 0.05). In primary biliary cirrhosis I-IV caffeine clearance is higher than in controls (117 +/- 59 ml/min, p < 0.05). In cirrhotic liver disease of different origin caffeine clearance is inversely related to the serum bilirubin level. However, the absolute value is determined in addition by the underlying disease.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7483717 TI - [Toxic alcohol-induced, acute intrahepatic cholestasis with reversible increase of the CA 19-9 tumor marker in chronic alcoholic hepatitis]. AB - The case report on an acute intrahepatic cholestasis caused by alcoholic intoxication in a 52-year-old medical laboratory assistant is given. In addition to extreme pathologic cholestasis parameters the increase of the tumor marker CA 19-9 was remarkable. By means of laparoscopy and histology, chronic fatty liver hepatitis due to alcoholic intoxication was found. The tumor marker CA 19-9 returned to normal following abstinence from alcohol parallel to the deline of the cholestasis parameters, which means the diagnostic importance of tumor markers in diseases with intrahepatic cholestasis is to be seen in relative terms. PMID- 7483718 TI - [Differential liver siderosis diagnosis using quantitative iron determination in the biopsy specimen]. AB - Quantitative iron investigations were carried out in the leavings of liver biopsy specimens from 92 patients with liver siderosis, detected by histochemical methods. A discrimination between primary and secondary liver siderosis were tried by the liver iron index. Idiopathic hemochromatosis was not found in liver iron indices below 2, both hemochromatosis and secondary liver siderosis were found with an index above 2. Only idiopathic hemochromatosis was found with an index above 5 with the exception of one transfusional siderosis in refractory anemia. PMID- 7483720 TI - [Histological changes in liver tissue in cholestasis]. AB - Cholestasis is a combination of defined clinical, paraclinical and morphological findings of which icterus is the direct symptom. It is the consequence of the delay or detention of the bile flow. By means of light microscopy different changes can be observed in hepatic tissue: formation of biliary thrombi, intracellular deposition of biliary components, ductal cholestasis with dilatation of the bile-duct lumina and applanation of the bile-duct epithelium, ductular proliferates and furthermore inflammatory mesenchymal reactions with cholangiolitis and cholangitis. These changes strengthen in dependence on the intensity and duration of the existing cholestasis. Bile lakes, bile extravasates, and biliary infarctions may occur in the process. The differentiation between intrahepatic non-mechanically and (extra-)hepatic mechanically conditioned cholestasis is of clinical importance. In the former the production and secretion of bile is disturbed (hepatosis, hepatitis and others), in the latter the extrahepatic bile ducts are above all affected: the bile flow is disturbed by stenosis, compression and obturation. The differentiation between the two kinds of cholestasis is impossible in the first three weeks because both of them develop identical lesion patterns. Distinguishable changes occur only after that. Consequently, findings from liver biopsy can only be a statement of probability. Reliable differentiation between the two kinds of cholestasis is however possible by means of modern technical methods for diagnostic purposes. PMID- 7483719 TI - [The prognostic value of serochemical cholestasis parameters in liver cirrhosis]. AB - In a semiprospective study on the prognosis of liver cirrhosis we investigated whether common, selected cholestasis parameters in liver diagnostics are associated with the survival time and the probability of survival and whether they permit prognostic statements. In 93 patients with liver cirrhosis of different aetiology the serum-bilirubin, gamma-GT, AP and AAP were determined for this purpose and applied in the same way as a clinically proven icterus to the survival time of the patients. The mean time of observation was 13.8 years (+/- 1.9 years). In the simple correlation matrix none of the cholestasis parameters correlated with the survival time. There was no difference in the frequency with which deceased and surviving patients suffered from icterus and, with the exception of AAP, there was no different serum concentration of the cholestasis parameters according to univariate analysis. However, icterus in the early phase of the observation time and increased gamma-GT (over 3000 nmol/l.s) in the late phase of observation were associated with a reduced probability of survival. Correspondingly, the serum-bilirubin of deceased patients (only slightly) correlated in the simple linear regression with their survival time. In multivariate analysis gamma-GT proved to be non-redundant for the estimation of the patients' probability of survival. PMID- 7483721 TI - [Clinical significance of cholestatic viral hepatitis]. AB - Modern serologic methods permit the classification of the particular course a virus hepatitis takes into individual types of pathogenesis. This is the case with hepatitis A in which only cholestatic courses have been proved for sure. Cholestatic courses are observed in 5-10% of all cases of acute hepatitis A, with variations from country to country. The duration of the disease is considerably longer than in non-cholestatic hepatitis A. It is between 80 and 130 or even more days! The diagnostic difficulty consists in the clear delimination to other diseases, particularly to intrahepatic cholestasis by drugs or to posthepatic stenosis. Sonography and ERCP are useful technological methods in this situation. Specific therapeutical measures are not available due to the lack of knowledge of the pathogenesis of this type of acute virus infection. The prognosis of cholestatic hepatitis A is good. Short-term glucocorticoid therapy is recommended by some authors in long-term cases, which are associated with pruritus, general weakness, loss of weight and distinct icterus. The course of the disease is obviously not changed by this therapy, but the sometimes agonizing symptomatology is relieved. PMID- 7483722 TI - [Drug-induced cholestasis]. AB - Cholestasis is one kind of reaction of the liver to intolerable drugs. It is estimated that 2% of the patients who are treated for icterus in hospital suffer from drug intolerance. A differentiation is made between obligtory and optional liver-damaging substances. The latter are clinically important because they occur far more frequently. In principle nearly every drug may cause cholestasis, which must be taken into consideration especially with newly introduced drugs, e.g. in cholestasis caused by co-enzyme-A-reductase inhibitors in the treatment of hypercholesterinaemia. Cholestasis is most likely to occur after administration of the following group of substances: antiarrhythmics, antibiotics, tuberculostatics, salicylates, immunosuppressives, narcotics, tranquilizers, some antirheumatics, antidepressants, anticonvulsives, and sex hormones. This list does not claim to be complete or to be in any order of frequency. PMID- 7483724 TI - Unconjugated hyperbilirubinemia. Clinical, laboratory and metabolic aspects. AB - The definition of Gilbert's syndrome changed in the 1980s. Today this syndrome means a heterogeneous, relatively frequent condition that is caused by moderate, chronic, often familial non-conjugated hyperbilirubinemia with an increased bilirubin turnover in about 30-40% of the cases. PMID- 7483723 TI - [Cholestasis in imipenem/cilastatin treatment]. AB - The case report on a 22-year-old female patient shows how the treatment with Imipenan/Cilastatin resulted in a severe intrahepatic cholestasis. The serum bilirubin rapidly decreased after discontinuation of the antibiotic. The laboratory findings were no longer conspicuous four weeks after discharge from hospital. PMID- 7483725 TI - [Genetic polymorphism in Gilbert-Meulengracht syndrome (GMS)]. AB - N-Acetylation and debrisoquine hydroxylation phenotypes were determined in 54 patients with Gilbert's syndrome and in 247 (sulfamethazine) and 76 (debrisoquine) non-related healthy volunteers, respectively. 40 (74.1%) of the patients and 135 (54.7%) of healthy volunteers were slow acetylators (chi 2 = 6.87). In patients, the cumulative urinary excretion (CUE) of sulfamethazine (0-6 hours) was significantly reduced. No differences between the debrisoquine poor metabolizers were observed: Gilbert's syndrome 5/54 (9.3%), healthy volunteers 5/76 (6.6%). The metabolic ratios were similar in both groups as well as the CUE of debrisoquine and its metabolite. Gilbert's syndrome seems to be related in some way to N-acetylation but not to debrisoquine hydroxylation polymorphism. PMID- 7483727 TI - [Cholestatic erythrohepatic protoporphyria: porphyrin metabolism before and after liver transplantation]. AB - The ferrochelatase deficiency in protoporphyria leads to accumulation of protoporphyrin in erythrocytes and liver. Consequences are protoporphyrinemia with photosensitivity and liver damage (fibrosis, cirrhosis) with cholestasis. The latter are unpredictable and can be observed in about 10% of the patients. Protoporphyrin, the physiological main component of hepatocellular porphyrins, has a hepatotoxic effect in the high-concentrated crystalline storage form. The obligatory hepatobiliary excretion of the lipophil, erythropoietic increased accumulating protoporphyrin in protoporphyria strains the excretory function of the liver. Its restriction is followed by an exzessive protoporphyrin accumulation, which leads to protoporphyrin-induced, progressive cholestatic cirrhosis, icterus, and aggravation of the extrahepatic protoporphyrinemic cutaneous manifestation. In case of hepatobiliary complications a coproporphyria of diagnostic relevance develops with inversion of isomers. Simultaneously, the fecal protoporphyrin excretion decreases. After liver transplantation hyperbilirubinemia, protoporphyrinemia and coproporphyrinuria significantly went down. A protoporphyrinemia of about 20% of preoperative values reflects the persisting hereditary enzyme defect and the continuity of the metabolic disease. PMID- 7483726 TI - [Crigler-Najjar syndrome type II in a 17-year-old girl]. AB - The Crigler-Najjar-Syndrome type II is characterised by the decreased activity of the bilirubin-UDP-glucuronyltransferase. This fact leads to an increase of the indirect bilirubin to 6-10 mg/dl. The mode of hereditary transmission is autosomal dominant with variable penetrance. The age of manifestation varies from the first year to the second decade of life. The Crigler-Najjar-Syndrome type II of a 17-year-old girl is reported. The diagnosis was made on the basis of the pattern of laboratory parameters in combination with an uneventful morphology of the liver. Under treatment with Chlofibrat the decrease of the serum total bilirubin from 10.4 mg/dl to 2 mg/dl was achieved. PMID- 7483728 TI - Current problems in primary biliary cirrhosis. AB - Despite the many advances made in the field of PBC, the central questions of this enigmatic disease remain unanswered. The pathogenesis is uncertain and there is no effective therapy. Of the various aetiologies proposed for the disease none has really withstood the test of time: if the disease were truly auto-immune, it might be anticipated that there would be a clear-cut response to cortico-steroid therapy: although such treatment has a slight effect on improving liver chemistry, compared with other auto-immune diseases such as auto-immune chronic active hepatitis. Steroids are relatively ineffective. An infectious aetiology has been suggested but no organism has been reliably isolated or implicated in the disease. Attempts at treatment have been largely hindered by the long natural history and the uncertainty as to choosing appropriate endpoints for studies. Perhaps the most intriguing observation has been the association with the anti mitochondrial antibodies. While the antigens have been identified, these are ubiquitous enzymes locate within the mitochondrion. How antibodies to such antigens are involved in the pathogenesis remains uncertain although our recent demonstrations that some antigens may be present on the surface of biliary epithelial cells may answer the question. However, further work may be required to confirm these findings and then to determine the reasons why there is such abnormal antigen expression in the disease. PMID- 7483729 TI - [What is the onset of primary biliary liver cirrhosis?]. AB - The medical records of 49 consecutive patients with primary biliary cirrhosis of the liver were screened for informations about medical examinations during the years before the diagnosis was established. In 15 cases previous medical reports could be found. Evidence of liver disease (slight elevation of transaminases and gamma-GT) was documented up to 18 years before the diagnosis was proven. In 6 patients liver biopsies had been performed: normal 1 x, fatty liver 1 x, fibrosis 1 x, non-specific hepatitis 1 x, chron. pers. Hep. 2 x. The characteristic increase of alkaline phosphatase often occurred within a few months. Antimitochondrial antibodies became positive independent of the beginning of cholestasis. It can be concluded that early stage primary biliary cirrhosis must be considered in patients with long standing slight elevation of liver enzymes even without cholestasis when other causes can be excluded. PMID- 7483730 TI - Prolongation of the QTc-interval reflects the severity of autonomic neuropathy in primary biliary cirrhosis and in other non-alcoholic liver diseases. AB - It was the aim of our study to prove a potential correlation (a) between laboratory findings of cholestasis and autonomic neuropathy (AN) and (b) between the severity of AN and the prolongation of the corrected QT-time (QTc). The five standard tests of autonomic cardiac neuropathy were investigated. QTc was calculated according to Bazett's formula. 12 out of 14 patients with primary biliary cirrhosis, 18 out of 21 patients with HBsAg positive liver diseases and 11 out of 14 patients with cirrhosis of other origin had AN. No significant correlation between the laboratory parameters of cholestasis and AN was found. Abnormal QTc values (> 440 m/sec) were observed significantly more often (p < 0.002) in patients with AN than in patients without AN. Significant linear regression (p < 0.01) could be confirmed between the prolongation of the QTc-time and the severity of AN. Besides the non-invasive investigation of the cardiovascular reflexes the evaluation of the QT-time might be an additional diagnostic means to identify patients with an increased cardiovascular risk in chronic non-alcoholic liver diseases. PMID- 7483731 TI - [Infiltrating T-cells in primary biliary cirrhosis recognize mitochondrial epitopes]. AB - The primary biliary cirrhosis (PBC) is characterized by lymphoid infiltrates in the portal tracts of the liver and occurrence of antimitochondrial autoantibodies (AMA) in serum directed against the pyruvate dehydrogenase complex or other enzyme complexes that are located on the inner mitochondrial membranes. The destruction of the biliary tracts is thought to be mediated by autoreactive liver infiltrating T-cells. In this study we demonstrate the reactivity of peripheral and liver-infiltrating T-cells against a crude preparation of human liver subcellular fractions measured by the tritium-thymidine uptake. Peripheral blood mononuclear cells (PBMC) from 13 of 15 patients with PBC and from 3 of 9 patients with chronic autoimmune hepatitis recognized human liver mitoplasts, i.e. mitochondria depleted of their outer membranes. PBMC from patients with chronic viral hepatitis B and C or with extrahepatic cholestatic icterus and PBMC from healthy controls did not recognize this antigen. Monoclonal antibodies against HLA-class II molecules blocked this proliferative response. Clonal analysis of 115 liver-infiltrating T-cells derived from two diagnostic liver biopsies of patients with PBC revealed a predominance of activated CD4+CD8- T helper cells. Six CD4- positive T-cell clones were reactive to the HLM preparation when the antigen was presented by autologous B cell lines. MAb against HLA-class II or HLA DR inhibited the response whereas mAb against HLA-DP did not. These clones did not respond to other subcellular fractions of human liver tissue. We conclude that among liver-infiltrating T-cells in PBC autoreactive T-cells exist that recognize some epitopes on the inner mitochondrial membranes.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7483732 TI - Helicobacter pylori-associated gastritis: does it play a role in functional dyspepsia? AB - OBJECTIVES: The goal of this study is to averiguate whether Helicobacter pylori (Hp)-associated gastritis is involved in the etiology of functional dyspepsia (FD) or, at least in a subset of these patients. METHODS: 103 consecutive FD patients were prospectively analyzed and subdivided according to their main complaints into ulcer-like FD (n = 31), dysmotility-like FD (n = 24) or nonspecific FD (n = 48). After endoscopy with gastric biopsy, gastritis and Hp infection were assessed and graded by histology according to the Sydney System. The prevalence of gastritis, Hp and disease activity were analyzed among the three FD subtypes and against 42 age, sex and race matched controls. RESULTS: Hp was identified in 64 (62%) of the FD group and in 23 (55%) of the controls (p > 0.20, NS). We neither observed any increased prevalence of the infection among FD subgroups (dysmotility-like 58%, ulcer-like 58%, unspecified 67%), nor in the activity of gastritis among FD subgroups and controls. We conclude that Helicobacter pylori gastritis per se is not associated with functional dyspepsia. PMID- 7483733 TI - [Prognostic relevance of hepatitis C virus genotype for responsiveness to interferon-alpha]. AB - The influence of hepatitis C virus (HCV)-genotypes on the responsiveness to interferon- (IFN-alpha) was studied in 116 patients with proven chronic hepatitis C. 88 of 116 (76%) patients were infected with HCV-genotype 1, 7 (6%) with HCV genotype 2, and 21 patients (18%) with HCV-genotype 3. All patients received at least 3 MU recombinant IFN-alpha-2a, 2b or lymphoblastoid IFN-alpha tiw for at least 6 month (total IFN-alpha dose per patient 216-720 MU, mean 360; treatment duration 6-12 month, mean 8). The follow-up after cessation of therapy was 9-48 months (mean 25). Sustained normalization of the aminotransferase levels was observed in 20 (17%) of the 116 patients. 10 of the 88 (11%) patients with HCV genotype 1, 7 of the 21 (33%) patients with HCV-genotype 3 (p < 0.02), and 3 out of the 7 patients with HCV-genotype-2- infection achieved a sustained remission. No response was observed in 58 (66%) and 3 (14%) patients with HCV-genotype 1 and 3 infections, respectively (p < 0.002). All but one of the sustained responders remained HCV-RNA negative during the entire follow-up. There were no significant differences between the sustained responders and the group of non-responders and responders with relapse in relation to age, pretreatment aminotransferase levels, histological activity index, or given IFN-alpha dosage. HCV-genotyping is helpful in evaluating the responsiveness to IFN-alpha and will be of importance considering the indication of therapy. PMID- 7483734 TI - [Do proton pump inhibitors after endoscopic control of acute ulcer hemorrhage have an advantage over H2 receptor antagonists?]. AB - During a two year period (1992-1993) we investigated whether or not, after endoscopic therapy of bleeding ulcers, the suppression of gastric acid secretion with an administration of a proton pump blocker (Omeprazol) is more effective than the administration of H2-receptor antagonist (Ranitidin) with respect to prevention of recurrent bleeding episodes, frequency of surgical intervention and mortality. 106 patients (64 men, 42 women) were treated with the proton pump blocker and 126 patients (82 men, 44 women) received the H2-receptor antagonist. Patients were treated either with an initial dose of 80 mg Omeprazol followed by 3 x 40 mg Omeprazol i.v. or with a daily dose of 3 mg/kg body weight Ranitidin i.v. No significant differences could be detected between the two treatment regimes with respect to the parameters mentioned above. Rebleeding which could be controlled by endoscopic hemostasis occurred in 19.8% vs. 17.5% (Omeprazol/Ranitidin) of patients. Surgical intervention because of rebleeding was necessary on 8.5% vs. 8.7% of the patients. Mortality due to hemorrhage was 5.7% vs. 4.0%. From these results we conclude that, following endoscopic hemostasis of bleeding ulcers, Omeprazol has no advantage over Ranitidin using our dosage regimes. PMID- 7483735 TI - [Percutaneous transhepatic cholangioscopy (PTCS)--an important supplement in diagnosis and therapy of biliary tract diseases (indications, technique and results)]. AB - In 39 patients with biliary disease inappropriate for a transpapillary access (21 m, 18 f, mean age 62 y.) 61 percutaneous transhepatic cholangioscopies (PTCS) were performed. In 28 cases (71.8%) diagnostic PTCS was done in order to investigate the etiology of a biliary stenosis. Nine of these patients had histological as well as clinical outcome indicating a benign lesion. Histology was positive for malignancy in 14 (82%) out of 17 patients with clinically assumed malignant stenosis. The etiology of the stenosis remained obscure from a clinical point of view in two cases with negative histology for malignancy. In twelve patients (30.7%) we performed PTCS for endoscopically controlled laser or electrohydraulic lithotripsy of bile duct stones. The procedure was successful in all patients and stone fragments were delivered to the duodenum by saline lavage. Seven of these patients had stones combined with a benign stenosis and were additionally treated by long-term drainage (Yamakawa drain) for three to twelve months. Two of the seven patients had no relaps of the biliary stricture after a follow-up of ten and twelve months respectively. In five cases (12.8%) PTCS controlled insertion of a guide-wire was attempted after failure of the radiologically guided internalization of the percutaneous drainage. The procedure was successful in three of these patients. PMID- 7483736 TI - [Tuberculous anal fistula in acquired immunologic deficiency syndrome]. AB - We report here on a 36-year old, HIV-positive patient, who was sent to hospital with an anal fistula. A short time later during the course of an extensive diagnosis the anal fistula was recognized as an extrapulmonary manifestation of a miliary tuberculosis stemming from an immunodeficiency syndrome. A rapid conversion of the sputum, a normalization of the radiological findings and the absence of relapse are the results of the classic systemic fourfold therapy with myambutol, isoniazid, rifampicin and streptomycin. The danger of overlooking the fact that an anal fistula can be the clinically primary manifestation of a tuberculosis and the problems of a mixed infection within the scope of the acquired immunodeficiency syndrome are discussed. Tuberculosis as a frequent complicating infection of HIV-positive patients--often diagnosed some time before the AIDS-infection as in our patient--can be successfully cured by a high dose of intravenous pharmacotherapy, even when additional complications (parasitic stomatitis, increasing deterioration of the immunological parameters) are present. In order to show the large spectrum of the problems involved in the diagnosis, the therapy and the course of the active acquired immunodeficiency syndrome, we have focused here on the detailed description of the case report. PMID- 7483737 TI - [Bouveret syndrome--a rare gallstone complication]. AB - A 62 year old woman presented with the symptoms of a gastric outlet obstruction together with severe metabolic hypochloremic alcalosis. A gallstone in the duodenum with gastric outlet obstruction was diagnosed by abdominal ultrasonography. In a one-stage surgical procedure, cholecystectomy, enterolithotomy and repair of the cholecytoduodenal fistula were performed. The postoperative course was uneventful. Pathogenesis, clinical findings, diagnostic procedures and therapeutic options of gastric outlet obstruction secondary to a gallstone impacted in the duodenal bulb (Bouveret's Syndrome) are discussed. PMID- 7483739 TI - [Malignant lymphomatous polyposis--a rare disease entity of malignant centrocytic lymphoma with characteristic endoscopic appearance]. AB - It is considered as special gastrointestinal manifestation on the nodal centrocytic lymphoma of intermediate malignancy. We demonstrate a 77 years old male patient, who underwent colonoscopy because of rectal bleeding. We found the colonic mucosa dotted with small, flat, lentiform polyps. In the terminal ileum these polyps were much larger. No changes were seen in the esophagus. In the antrum we found flat elevations and in the duodenum again lentiform nodules. Presenting in stage IV of non-Hodgkin's lymphoma the patient was treated with five cycles of combined chemotherapy with prednimustin and mitroxanthrone without side effects. Restaging showed distinct remission of the disease with significant reduction of the mucosal alterations. PMID- 7483738 TI - [Therapy refractory atypical polyarthritis and cryoglobulinemia in a patient with colon carcinoma and palliative intestinal bypass. Differential diagnosis: carcinoma-polyarthritis or bypass arthritis]. AB - The case of a 63 year old woman with mesenteric recurrence of a colonic carcinoma and infiltration of the duodenum is reported. To bypass duodenal stenosis a duodenojejunostomy was performed. Three months later the patient developed severe atypical polyarthritis which led to hospitalization. The arthritis affected large and small joints in an asymmetrical pattern. Fever and Raynaud's phenomenon of both hands accompanied the arthritis. Elevated sedimentation rate, acute phase proteins, cryoglobulinemia and immune complexes were remarkable laboratory findings. Rheumatoid factor was absent. In the subsequent course the polyarthritis was refractory to steroids and nonsteroidal anti-inflammatory drugs. Only treatment with broad-spectrum antibiotics ameliorated the arthritis. Postenteric reactive arthritis, septic arthritis and metastatic arthritis could be excluded. Although the patient had a family history of rheumatoid arthritis and a HLA-type DR4 the diagnosis of rheumatoid arthritis was not very likely since distal interphalangeal joints were affected, rheumatoid factor was absent and antibiotic therapy was successful. The case serves to discuss carcinoma polyarthritis and bypass-arthritis as the main differential diagnosis. PMID- 7483740 TI - [Lipid island (xanthelasma) of the duodenal mucosa--a rarely found diagnosis and differential diagnosis]. AB - A case report ist given on a 54-year-old patient with the so called lipid island (xanthelasma) in the duodenal mucosa. Xanthelasmas are very rarely located in the duodenal mucosa. The diagnosis and differential diagnosis of this entity are reported. The relevance of histochemistry and immunohistochemistry for differentiation of signet ring carcinoma cells in the mucosa is discussed. PMID- 7483741 TI - [Diagnosis and therapy of autoimmune hepatitis]. AB - Thorough differential diagnosis in patients with autoimmune hepatitis is important since other liver disorders need different treatment regimen. Elevated transaminases and gamma-globulines and autoantibodies should make one think of autoimmune hepatitis. Liver biopsy is helpful but usually not a definitive proof of the diagnosis. Immunosuppressive therapy has to be administered for years and possibly leads to serious side effects. Therapy should only be discontinued if a complete remission is induced. Relapses after cessation of medical treatment occur frequently. If no remission is achieved lifelong immunosuppressive therapy has to be given. In decompensated disease liver transplantation offers a treatment with good prognosis. PMID- 7483742 TI - [Long-term prognosis after liver transplantation for hepatitis C virus-induced liver disease]. PMID- 7483743 TI - [Administration of acetylsalicylic acid and risk of colorectal adenoma and carcinoma]. PMID- 7483744 TI - 7th International Symposium Wilson's Disease. Baden/Vienna, Austria, 25-27 August 1995. Abstracts. PMID- 7483745 TI - [Cardiology 1995: Diagnosis and Therapy of heart diseases, current standards and clinical questions. 19th Autumn session of the German Society of Cardiology- Heart and Cardiovascular Circulation Research. Bielefeld, 21-23 September 1995. Abstracts]. PMID- 7483746 TI - [Cognitive capacity in advanced age: initial results of the Berlin Aging Study]. AB - This study reports data on intellectual functioning in old and very old age from the Berlin Aging Study (N = 516; age range = 70-103 years; mean age = 85 years). A psychometric battery of 14 tests was used to assess five cognitive abilities: reasoning, memory, and perceptual speed from the broad fluid-mechanical as well as knowledge and fluency from the broad crystallized-pragmatic domains. Cognitive abilities had a negative linear relationship with age, with more pronounced age based reductions in fluid-mechanical than crystallized-pragmatic abilities. At the same time, ability intercorrelations formed a highly positive manifold, and did not follow the fluid-crystallized distinction. Interindividual variability was of about equal magnitude across the entire age range studied. There was, however, no evidence for substantial sex differences. As to origins of individual differences, indicators of sensory and sensorimotor functioning were more powerful predictors of intellectual functioning than cultural-biographical variables, and the two sets of predictors were, consistent with theoretical expectations, differentially related to measures of fluid-mechanical (perceptual speed) and crystallized pragmatic (knowledge) functioning. Results, in general indicative of sizeable and general losses with age, are consistent with the view that aging-induced biological influences are a prominent source of individual differences in intellectual functioning in old and very old age. Longitudinal follow-ups are underway to examine the role of cohort effects, selective mortality, and interindividual differences in change trajectories. PMID- 7483747 TI - [Recall of personal performance in the elderly]. AB - One shortcoming of traditional memory research with older adults is that the scope of memory tasks and memory materials used has been rather narrow. The studies to be presented in this paper, assessing memory for self-performed action events, were designed to answer the question whether the well-documented age related memory decline indicates a global characteristic of the aging memory system or if this decline is specific to the verbal memory system. It could be demonstrated that memory deficits older adults show when learning and remembering self-performed action events are similar to those that have been found many times for verbal materials. Thus, the age effect in memory is not limited to verbal materials. PMID- 7483749 TI - [Integration of prior knowledge in speech comprehension in relation to age]. AB - In a first study we confirm the general evidence concerning the influence of implicit knowledge structures on sentence and text comprehension. In addition the results suggest a differentiation in the availability of prior knowledge. The level of activation is dependent on the type of semantic relations between prior knowledge and the explicit sentence information. This result can be explained by a theoretical framework in which different levels of cognitive effort can be related to activation or production of such relations. The main point of our second experiment is to prove the influence of aging on the process of knowledge integration mentioned above. We compared the results of a subject group of young (25 years) with old adults (75 years) in a recognition task. The experimental data support a slowdown for primarily sensorical and motorical and nonlexical components for the higher age group but they do not confirm a deficit in the critical process of knowledge integration. Finally we discuss a differentiation of such integration procedures dependent on demand of memory capacities (activation vs. operations) and the advantage of such a classification on identification of specific cognitive deficits. PMID- 7483748 TI - Analyses of adult age differences in associative learning. AB - Data from a recent project concerned with adult age differences in associative learning were reanalyzed to identify the processes involved in the age-related differences in trial-by-trial performance in this task. A measure reflecting the failure to retain feedback relevant to one's previous response accounted for a large proportion of the age differences in the early trials of associative learning. The speed with which individuals completed simple processing operations also contributed to the age differences both in measures of accuracy (percentage correct) and in measures of feedback retention. PMID- 7483750 TI - [A model of the population growth of the Earth]. AB - Of all global problems world population growth is the most significant one. World population drives all other global issues--of food and energy, resources and the environment. Although demographic data in a quantitative way does express the net result of our economic, social and cultural development, methods of demography can hardly suggest a more general, synergistic, approach to the study of long term growth. To describe this process in its past and project it into the foreseeable future a mathematical model is worked out. It treats the world population as an entity, seen as an open and evolving dynamic system. The approach is phenomenological and growth over very many generations is assumed to be self-similar. This invokes scaling and mathematically is expressed by a power law. Comparison with population data leads to a consistent description of growth by a hyperbolic growth curve valid from the Paleolithic up to the middle of this century. In terms of kinetics the growth rate is proportional to the square of the total number of people, rather than their number, as in the case of exponential growth. Thus nonlinear quadratic rate describes the sum outcome of all mechanisms that contribute to our growth and development in a collective interactive process. Taken without limits this pattern is divergent in the distant past and present, when it describes the population explosion. These singularities are cut off by introducing a characteristic time of 42 years,--the effective human life span--as the microscopic time constant in this phenomenology. The model gives an estimate of the beginning of human evolution congruent to 4.4 million years ago and of the total number of people who ever lived: congruent to 100 billion. In the scope of the model large scale cycles defined by history and anthropology are shown to be uniformly spaced in time on a logarithmic scale, expressing an inherent periodicity. On this scale, as we go into the past the rate of growth decreases when, finally in the early Paleolithic any appreciable change would take a million years to happen. As we approach the present this progression of cycles with the consequent compression of the time scale is now terminated by the demographic transition. This is seen as a singular epoch in all human development when the basic mode of world population growth is to change.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 7483751 TI - [The regulation of calcium homeostasis in nerve cells]. AB - We have reviewed the major cellular elements related to the release and buffering of calcium in neurons. Voltage-operated, chemical-operated calcium channels and mechanisms of stability of intercellular calcium homeostasis (mitochondria, endoplasmic reticulum, calcium binding proteins, calcium exchange and calcium pump) are demonstrated in normal and pathological condition (105 ref.). PMID- 7483752 TI - [The epiphysis in the visual analyzer system]. AB - The brain gland epiphysis, participated in the regulation of the endocrine functions of mammals, is rudimentary third eye of lower vertebrates. In the process of evolution the gland has lost it own photoreceptor functions, but saved very close contacts with visual system. By means of it biologically active combinations and, first of all, melatonin hormone, the epiphysis support control over transmission of visual information and participate in realization of biological effects of light on organism. PMID- 7483753 TI - [The physiology of the kidney and water-salt metabolism: the basic research trends in Russia]. AB - The major tendency for the investigations of kidney function and water-salt metabolism running in different Russian laboratories is reviewed. The proceedings of some national publications on the regulation of water and ion transport at the cell and molecular level demonstrate the progress in this field of renal physiology in Russia. Trends in the researchers of the renal function and its neuroendocrine regulation on experimental animals and also on people with renal dysfunction are described. In the last part of the review there is a brief account of the papers on the development of the kidney function and water-salt metabolism during onto- and phylogeny. PMID- 7483754 TI - [The mechanisms of synaptic plasticity: the role of synaptic protein phosphorylation and gene expression]. AB - Results of investigations of protein phosphorylation and gene expression in the mechanisms of long-term potentiation and kindling was analyzed. It was discussed the participation of N-methyl-D-aspartate receptors, intracellular Ca2+, different protein kinases and immediate-early genes in the induction and maintenance of long-term changes of glutamatergic synaptic efficiency in hippocampus. It was suggested the essential role of the level of intracellular Ca2+ in the induction of the cell program leaded to the trace processes during electrical stimulation of the same hippocampal area. PMID- 7483756 TI - [The functional organization of the spatial structures of the neuronal receptive fields in field 21 of the cat cerebral cortex]. AB - In result study of structural organization of neural receptive fields (RFs) and their different zones on the prestriate cortex level was shown that neural RFs in this area have hypercomplex structure and consist of some spatial different excitatory zones. Orientation and velocity selectivity and spatial-frequency characteristics of the excitatory zones of the same RF may differ between zones. The number of zones in RFs correlate with RF sizes: the more RF size the more zones number in them (r = 0.05, P < 0.02). About 66% of zones in RFs have approximately identical sizes (12-16 deg) and sizes those zones an dependence as from their number in RFs (r = 0.03, P > 0.05) as from the eccentricity (r = 0.3, P > 0.05). Zones in RF were distributed so that distance their centres was between 56-65 degrees (an average across all 60 degrees). If two or three zones were tested simultaneously the neuron changed own frequency and orientation tuning. Consequently the functional organization of neural RFs of prestriate cortex depend upon number simultaneously tested zones in spatial information processing. Was shown that important properties of prestriate cortex neurons is their ability to integrate and complex processing of spatial information across wide area of the visual field; in the prestriate cortex thus violate of straight retinotopical representation principle of visual field, characteristics of all investigated areas of visual system. The results represented in this review allow suppose that the structural and functional unit of cortex which make integration of spatial information from different compose this information is prestriate cortex neurons and not is no striate cortex modules. Consequently combined of neurons which make images description from different parts of visual field out of dependence of cortical area in turn integrate in neuron networks. Is known that neurons of any level have characteristics different of (from) characteristics of other levels visual system. The interaction of different elements of visual system form ultrastructure of RFs and their different zones in prestriate cortex, and ensure function of networks. So just on prestriate cortex neurons level appear whole system of surrounding world universe system of connections from different cortical areas which is morphological and functional base in process of spatial information integration RFs which have the central and peripheral representation. PMID- 7483755 TI - [The factors supporting peripheral nerve regeneration]. AB - At present work the special mechanisms for stimulating influence of the growth cone mobility extracellular regulators upon peripheral nerve fibers regeneration were passed in review. An important role played by the system of neuron-Schwann cell informational interactions and the GAP-43 neurospecific protein, that is known to affect specifically the axonal growth cone while nerve fibers regeneration, was mentioned. There's considerably less data available on the grave problem of physical factors and various neuroplastic approaches influence upon nerve regeneration. Investigations in such a field are characterized with absence of profound comprehension of the cell-cell as well as cell-molecular interactional mechanisms observed within nerve regeneration. Nevertheless, those mechanisms consideration in our article seems us to be rather helpful for the reason of the generally taken into account further acquirements of the new data on the role played by the diffusible molecular signals, adhesion molecules and extracellular matrix during the nerve fibers regeneration. PMID- 7483757 TI - [Methods for analyzing brain evoked potentials clinically and in psychophysiology]. AB - In the article one can find compare classification of excited brain potential analysis methods using for tasks of neurophysiology of man. Importance of visual data analysis, methods of multimeasure statistics and correlation analysis are discussed. Methods of dipoles localization and three-channel "lissa-djouson" trajectory are analysed from the position, that head is volume conductor. The last part of the article is devoted to importance of space methods of analysis of excited potentials, mainly for mapping of bio-electrical and magnetic exited potentials. PMID- 7483758 TI - Synthetic peptide vaccines: success at last. PMID- 7483759 TI - Oral immunoadjuvant activity of lipophilic derivatives of N-acetylglucosaminyl beta(1-->4)-N-acetylmuramyl-L-alanyl-D-isoglutamin yl-(L)- 2,6-meso diaminopimeric acid-(D)-amide. AB - Twenty-four kinds of the acylated, amidated or esterified derivatives of N acetylglucosaminyl (GlcNAc)-beta(1-->4)-N-acetylmuramyl (MurNAc)-L-alanyl (Ala)-D isoglutaminyl (isoGln)-(L)-meso-2,6-diaminopimeric acid (A2pm)-(D)-amide (GMP3-A) which were prepared by chemical modifications of an enzymatic hydrolysate of Lactobacillus plantarum cell-wall peptidoglycans were examined for oral adjuvant activity by gastric intubation with bovine serum albumin (BSA) in liposomes into BALB/c mice. The gastric intubation of GlcNAc-beta(1-->4)-MurNAc-L-Ala-D-isoGln (L)- stearoyl-(D)-meso-A2pm-(D)-amide-(L)-O-octyl, -nonyl and -dodecyl esters exhibited the most marked oral adjuvant activity in terms of enhanced production of serum anti-BSA IgG antibody. Some derivatives showed a less marked adjuvanticity and others were totally inactive. Thus the oral adjuvanticity of test lipophilic derivatives of GMP3-A in liposomes was dependent upon their chemical structure. PMID- 7483760 TI - Safety, immunogenicity, and kinetics of the immune response to a single dose of virosome-formulated hepatitis A vaccine in Thais. AB - The safety and immunogenicity of a single dose of virosome-formulated hepatitis A vaccine was evaluated in healthy seronegative Thai volunteers. Immunization elicited primarily mild transient reactions which did not interfere with normal activities. All subjects possessed > 20 mIU of anti-hepatitis A virus antibody per ml of serum one month after immunization. Such a prolonged rise in antibody titre is characteristic of virosome-formulated vaccines. Protective titres (> 20 mIU ml-1) were maintained by all subjects over the one-year observation period. PMID- 7483761 TI - The simian immunodeficiency virus transmembrane protein is poorly immunogenic in inactivated virus vaccine. AB - The transmembrane proteins (TMP) of immunodeficiency lentiviruses are primary candidates for inclusion in AIDS vaccines, the design and testing of which is facilitated by the SIV-macaque infection model. Antibody responses to linear determinants in the SIVmac TMP were investigated in rhesus macaques either infected with the SIVmac J5 molecular clone or vaccinated with partially purified, formalin-inactivated SIVmac. Infected animals were shown to recognise predominantly four regions in the external domain and three regions in the internal domain of the TMP defined by a series of nominally 20mer overlapping peptides. In contrast SIV vaccinates had extremely restricted and weak antibody responses to the TMP, indicating a selective loss of immunogenicity of this component in the vaccine. PMID- 7483762 TI - Intestinal immunization of mice with antigen conjugated to anti-MHC class II antibodies. AB - We have explored a new technique for immunization of the intestinal tract of mice, using protein antigens bound to antibodies with specificity for murine MHC class II molecules (MHC-II). Either of two protein antigens, hen avidin (AV) or hen egg lysozyme (HEL) were covalently conjugated to anti-MHC-II antibodies and the purified conjugates were given orally (p.o.) or by direct intraduodenal (i.d.) injection into the intestinal lumen of mice. A secondary immunization p.o. with the same conjugate or with the non-conjugated antigen in the presence of cholera toxin (CTX) resulted in production of both intestinal secretory IgA and serum IgA antibody by those mice. In addition, serum IgG antibodies were produced. Conjugates with appropriate MHC-II specificity targeted the antigen because they induced more IgA and IgG antibody than conjugates with irrelevant antibody specificity or antigen alone, and because they induced antibody in mice that were genetic low responders to antigen. The results indicate the feasibility of oral subunit type vaccines with antibody targeting technology. PMID- 7483763 TI - Immune response of calves to foot-and-mouth disease virus vaccine emulsified with oil adjuvant. Strategies of vaccination. AB - Calves born to vaccinated cows under the regular annual vaccination programme were vaccinated at different ages using commercial quadrivalent (01, A79, A87 and C85 FMDV strains) vaccine emulsified in oil adjuvant. The antibody responses of vaccinated calves were evaluated using liquid-phase blocking sandwich ELISA. All calves 20, 30 and 40 days old having high maternal antibody titres responded well to vaccination. Moreover, 25-57% of vaccinated calves showed protective antibody titres both at 90 and 120 days post-vaccination (d.p.v.), whereas none of the non vaccinated animals achieved these levels. Calves aged 3-4 months with non protective levels of colostral-derived antibodies responded with high antibody titres to vaccination which persisted for at least 4 months. In both groups of calves a certain degree of suppression of postvaccinal response was observed which was related to colostral antibody titres. Our results suggest that in order to reduce the proportion of calves susceptible to infection it is advisable to immunize calves as young as 20 days old to induce acceptable antibody titres for the following 4 months. In addition, a second vaccination 60 d.p.v. ensures high antibody levels in high disease risk areas. PMID- 7483764 TI - Comparison of the T helper cell response induced by respiratory syncytial virus and its fusion protein in BALB/c mice. AB - Specific proliferative T-cell responses were induced in the lymph node cells (LNC) of mice immunised with a sucrose density gradient purified preparation of respiratory syncytial (RS) virus or an immunoaffinity purified preparation of the F glycoprotein. Inhibition studies and flow cytometric analysis showed that the responding cell population were CD4+ T cells. The cytokines produced by virus specific and F-specific cells were assessed using the CTLL cell line. Peak quantities of cytokine were consistently detected in the supernatants of stimulated cultures 24 h prior to maximum proliferation. The proportion of IL-2 released was determined by blocking IL-2 activity with an anti-IL-2 monoclonal antibody. In cultures of RS virus primed LNC challenged with whole virus there was a switch of cytokine production from 70% IL-2 at day 3 to 80% IL-4 by 6 days of culture. In contrast, LNC cultures from mice immunised with F protein secreted 75-100% IL-2 throughout the culture period. These data suggest that after 6 days of challenge with viral antigen, the RS virus-primed LNC response consists of T helper cells which are predominantly of the Th2 subset, secreting IL-4, whilst F protein-primed LNC secrete large quantities of IL-2 and can therefore be classified as predominantly of the Th1 subset. PMID- 7483766 TI - Protection of mice against lethal viral infection by synthetic peptides corresponding to B- and T-cell recognition sites of influenza A hemagglutinin. AB - Previously, we reported 12 synthetic T- and B-cell recognition regions representing surface areas of the hemagglutinin (HA) of X31 influenza virus. In the present study, four of these peptides were examined in Balb/c mice for their ability to produce protective immunity against lethal infection with a dose equivalent to 10 LD50 of influenza virus. These peptides corresponded to the following sequences: 23-36 (HA1-1); 138-152 (HA1-3); 183-199 (HA1-6) and 1-11 (HA2-10). Each of the selected peptides, in their free form, evoked anti-peptide antibodies that cross-react with intact X31 virus. Two of the peptides, HA1-1 and HA1-3, also elicited virus-specific delayed type hypersensitivity (DTH) responses. These two peptides, when injected into mice, not only failed to protect the immunized mice against challenge with influenza virus, but in fact caused greater susceptibility to viral infection as compared to control animals that had been injected with saline. In contrast, peptides HA1-6 and HA2-10, which were unable to induce adequate virus-specific DTH responses, conferred 42-46% and 54-73% protection, respectively, compared to the control group that received only saline (P < 0.03 to P < 0.01). PMID- 7483765 TI - The efficacy of a live Listeria monocytogenes combined serotype 1/2a and serotype 4b vaccine. AB - In each of two experiments, sheep and lambs were vaccinated by a subcutaneous injection of a test vaccine (consisting of a combined serotype 1/2a and serotype 4b live Listeria monocytogenes culture) and challenged 16 days later with a mixture of the homologous wild strains. After challenge, the mortality rate of vaccinated sheep was 28.1% and that of controls 71.9%; that of lambs was 25.0%, although these had been inoculated with the LD70 dose. Furthermore, in each of two field trials in Listeria-infected flocks, primiparous pregnant ewes were vaccinated. In the first field trial, 3 or 110 lambs died of listeriosis of those born of vaccinated (n = 564) or unvaccinated (n = 3345) ewes, respectively. In the second, the perinatal mortality rate of lambs born of vaccinated or unvaccinated ewes was 7.6 or 30.3%, respectively, and the mean birth weight of lambs born of vaccinated or control ewes was 2.2 or 1.8 kg, respectively; the mean milk production of vaccinated ewes was 106 and that of controls 83 l; no Listeria was isolated from milk samples of vaccinated ewes. It is concluded that the vaccine is efficacious for the protection of sheep from listeriosis. PMID- 7483767 TI - Gene fusion of cholera toxin B subunit and HBV PreS2 epitope and the antigenicity of fusion protein. AB - A unique EcoRI site was introduced at the 3' end of cholera toxin B subunit (CTB) gene by site-directed mutagenesis, polynucleotides encoding 120-145aa epitope of HBV PreS2 were chemically synthesized and fused to the 3' end of cholera toxin B subunit gene. The fused gene was over-expressed (about 30 micrograms ml-1) in E. coli, and more than 95% of the fusion protein was secreted into the medium. The fusion protein expressed was purified by affinity chromatography. The chimera protein obtained bound to ganglioside GM1, and had the antigenicity of both cholera toxin B subunit and HBV PreS2 as confirmed by ELISA. After mice were immunized intramuscularly with the fusion protein, anti-CTB antibody and anti PreS2 antibody were produced. These results indicated that the fusion protein retained not only the biological function of CTB but also the antigenicity and the immunogenicity of cholera toxin B subunit and HBV PreS2 epitope. This work provided a sound basis for further studies on the construction of engineered peptide vaccine. PMID- 7483768 TI - Simultaneous expression of CFA/I and CS3 colonization factor antigens of enterotoxigenic Escherichia coli by delta aroC, delta aroD Salmonella typhi vaccine strain CVD 908. AB - Among the known colonization factors of enterotoxigenic Escherichia coli (ETEC), CFA/I and CS3 (the common antigen in the CFA/II family of fimbrial antigens) are two of the most prevalent fimbrial antigens found in clinical isolates but are never expressed by the same wild-type strain. We manipulated the genetic determinants encoding CS3 and CFA/I fimbriae so that these two important colonization factors are expressed simultaneously in attenuated Salmonella typhi live oral vaccine strain CVD 908, including after growth in liquid medium (CFA/I is poorly expressed by wild-type ETEC in broth culture). The recombinant fimbrial structures produced by CVD 908 are morphologically indistinguishable from the CS3 fibrillae and CFA/I rod-like fimbriae produced by ETEC, and are recognized by monospecific CS3 and CFA/I antibodies. This prototype construct may prove useful in investigating the live vector approach to immunoprophylaxis of ETEC diarrheal disease. PMID- 7483770 TI - Isotype profiles induced in Balb/c mice during foot and mouth disease (FMD) virus infection or immunization with different FMD vaccine formulations. AB - The IgG isotype response in Balb/c mice infected with FMDV or immunized with different vaccine formulations using inactivated virus particles as antigen was analyzed at various times post-inoculation. For this purpose an ELISA based on polyclonal antibodies for detection and quantification of mouse IgG isotypes with FMD virus (FMDV) specificity was developed. Three immunomodulators, which have been shown to be very effective in inducing strong and long-lasting antibody responses (Bahnemann, Arch. Virol. 1975, 47, 47-56; Polatnik and Bachrach, Appl. Microbiol. 1964, 12, 368-376), were employed to formulate different vaccines using aqueous and oil vehicles: a water-soluble fraction of the cell wall of Mycobacterium sp., a purified extract of lipopolysacharide from Brucella ovis and a synthetic lipoamide, Avridine. Infected animals between 14 and 60 days post inoculation (d.p.i.) showed responses dominated by IgG2b, followed by IgG1, IgG2a and IgG3, respectively. The IgG3 isotype was the first, together with IgG1, to be elicited during the first 7 days after infection, whereas no IgG3 activity was detected in vaccinated animals at any time. With formulations including immunomodulators, persisting high levels of IgG2b (similar to those of infected animals) were detected until 180 d.p.i., while with conventional vaccines IgG2b responses were detected up to 60 d.p.i. Animals vaccinated with formulations including these immunomodulators presented an augmented resistance to viral challenge at 210 d.p.i. in relation with those immunized with conventional vaccines. The possible relationship of these differences in the isotype response and protection is discussed. PMID- 7483769 TI - Immunogenicity of the recombinant serine rich Entamoeba histolytica protein (SREHP) amebiasis vaccine in the African green monkey. AB - We report the first study in non-human primates of the safety and immunogenicity of a recombinant vaccine designed to prevent amebic liver abscess. In a pilot study, a recombinant vaccine containing the serine rich Entamoeba histolytica protein (SREHP) attached to a maltose binding protein (SREHP/MBP), which has been shown to be effective in preventing amebic liver abscess in rodent models of infection, was used to immunize two African Green Monkeys. Vaccination with SREHP/MBP resulted in no systemic side-effects. The monkeys receiving the SREHP/MBP protein developed antibodies that recognized the recombinant SREHP/MBP molecule, the native SREHP protein, and the surface of amebic trophozoites. Antiserum from SREHP/MBP-vaccinated monkeys could block the adhesion of E. histolytica trophozoites to mammalian cells, a feature that may correlate with vaccine efficacy. Attempts to produce amebic liver abscess in naive African Green Monkeys by direct hepatic inoculation with virulent E. histolytica trophozoites was not successful, suggesting this species is probably not suitable for vaccine efficacy studies. PMID- 7483771 TI - Phylogenetic evidence of canine distemper virus in Serengeti's lions. AB - Recently an epizootic, reported to be due to a morbillivirus infection, affected the lion population of the Tanzanian Serengeti National Park. A morbillivirus phosphoprotein (P) gene fragment was amplified by PCR from tissue samples of several affected lions. Sequencing of the amplificates and subsequent phylogenetic analyses revealed that a wild-type strain of canine distemper morbillivirus (CDV) was involved. Vaccination of the local domestic dog population with proven safe CDV vaccines is proposed. PMID- 7483772 TI - Development of a guinea pig model to assess immunogenicity of Haemophilus influenzae type b capsular polysaccharide conjugate vaccines. AB - There is currently no animal model which reliably predicts the immunogenicity of Haemophilus influenzae type b (Hib) polysaccharide-protein conjugate vaccines in human infants. We evaluated various Hib vaccines in guinea pigs using techniques similar to the United States potency test for adsorbed diphtheria and tetanus toxoids with a view to developing a method for evaluating the potency of a combined adsorbed tetanus, diphtheria, pertussis and Hib conjugate vaccine. Groups of 6-8 guinea pigs received 1.5 single human doses of vaccine at 0 and at 6 or 8 weeks and were bled at 6 weeks and 2 weeks after the booster injection. Total antibodies to polyribosylribitolphosphate (PRP), the Hib capsular polysaccharide, were measured in individual animals and in serum pools by radioimmunoassay. The relative antibody responses of guinea pigs to Hib conjugate vaccines qualitatively resembled those of human infants. Unconjugated polysaccharide was not immunogenic; PRP-D produced a low antibody response, HbOC, PRP-T (Merieux) and Hib-T (MPMBL) produced a low response to the first dose and a strong anamnestic response to the booster (geometric mean anti PRP > 1 micrograms ml-1). PRP-OMP uniquely produced a strong response after the first dose which was further boosted by the second dose. Experimental Hib-T vaccine lots with low levels of conjugation were poorly immunogenic in guinea pigs. Combinations of DTP and Hib-T vaccines showed equivalent or greater immunogenicity than Hib-T alone. We propose that the guinea pig model may be useful to verify the immunogenicity of PRP conjugate vaccines and for pre-clinical evaluations of DTP-Hib combination vaccines containing PRP conjugates. PMID- 7483773 TI - MMR vaccination, measles epidemiology and sero-surveillance in the Republic of Ireland. AB - OBJECTIVE: Following the introduction of a national measles and subsequent MMR vaccination programme, to determine the susceptibility of 3-14-year-old children to measles, mumps and rubella and to relate the results to the epidemiology of measles and the need for vaccination policy changes. DESIGN: Cross-sectional sero survey and trends in measles notifications and mortality. SETTING: Paediatric hospital outpatient departments in Dublin. SUBJECTS: Sera were collected from 837 children attending the clinics in 1991 and 1992. RESULTS: The prevalence of antibodies in children aged 3-6, 7-10 and 11-14 years was 84, 83 and 95% for measles; 48, 60 and 65% for mumps; and 78, 63 and 74% for rubella, respectively. The prevalence of mumps antibodies may be underestimated. Ninety-six per cent of girls aged 13-14 years had rubella antibodies. A widespread outbreak of measles occurred in 1993. Over recent years, an increasing proportion of measles notifications were in older children. CONCLUSION: Given sub-optimal uptake of MMR vaccine, outbreaks of infection in pre- and primary school children are inevitable. In such circumstances, a 2-shot MMR vaccine programme with high uptake is essential to prevent a shift of disease into older age groups. PMID- 7483774 TI - Biological and immunogenic properties of a canarypox-rabies recombinant, ALVAC-RG (vCP65) in non-avian species. AB - A canarypox-based (ALVAC) recombinant expressing the rabies G glycoprotein has been utilized to assess in vitro and in vivo biological properties of the canarypox virus vector system. In vitro studies have shown that no replication of the virus can be detected on six human-derived cell lines, nor can the virus be readily adapted to replicate on non-avian cells. Expression of the rabies G can be detected on all cell lines analyzed in the absence of productive viral replication. Analysis of viral-specific DNA accumulation indicated that the block in the replication cycle in the human cell lines analyzed occurred prior to DNA replication. The exact nature of the block, however, remains unknown. The concept of using a non-replicating immunization vehicle has been demonstrated through extensive in vivo studies in a range of species including non-human primates and humans. The results of such in vivo studies have exemplified the safety and immunogenicity of the ALVAC vaccine vector. PMID- 7483778 TI - Hybrid genes over-express pertactin from Bordetella pertussis. AB - Pertactin is a surface adhesin of Bordetella pertussis which is produced in small quantities when expressed from the native prn promoter. Hybrid genes were constructed in which the prn promoter was replaced by either the fha or tox promoter. Recombinant B. pertussis strains containing chromosomally integrated hybrid tox promoter/prn (toxpprn) or fha promoter/prn (fhapprn) genes expressed pertactin at approximately 5- and 8-fold the wild-type level, respectively. The pertactin was correctly processed and secreted and was biochemically and antigenically comparable to its wild-type counterpart, as determined by N terminal sequence analysis, immunoblotting, peptide mapping, circular dichroism and antigenicity studies. In an adherence assay, a strain over-expressing pertactin was no more adherent than the wild-type strain, but a pertactin deficient strain was less adherent. PMID- 7483775 TI - Anti-HBV neonatal immunization with recombinant vaccine. Part I. Critical appraisal for a long-lived antibody course. AB - This study involved 912 infants born to HBsAg-negative mothers from 1 May 1991 to 30 June 1992. The subjects were randomly allocated to an accelerated (Group A) or traditional (Group B) immunization schedule and immunized with 10 micrograms of recombinant HBV vaccine. At the end of the vaccinal cycle 98.14% of both groups were protected against HBV with a high percentage of high responders (88.1% group B and 68% group A). Following a random plan, 345 of the initial 912 infants (144 group A and 201 group B) were serologically evaluated, 15-18 months after the booster dose, to identify the level of long-lasting specific antibody. The data obtained allowed us to identify the non-responder subjects after the seroconversion, to propose the evaluation of antibody titre after the booster dose of vaccine and, because one year after the booster dose 5.6% of the subjects responsive at seroconversion have shown undetectable anti-HBsAg titre, to propose the elevation of the antibody level considered as protective at the end of the vaccinal cycle. PMID- 7483777 TI - Construction and characterization of a Salmonella typhi-based human immunodeficiency virus type 1 vector vaccine. AB - Since the human immunodeficiency virus type 1 (HIV-1) is transmitted either parenterally or sexually, both systemic and mucosal immune responses might be required to provide protective immunity. One option is to express HIV proteins in attenuated Salmonella vectors that elicit immune responses in both compartments. The first step to constructing such a strain was achieved by integrating a gene expression cassette encoding recombinant HIV-1 gp120 (rgp120) into the aroC locus of an attenuated vaccine strain of S. typhi. This rgp120 expression cassette utilizes the strong constitutive promoter, P1pp/lacUV5, and produces rgp120 to 0.05-01% of the total bacterial cell protein. Immunoblot analysis shows that the S. typhi strains containing the integrated cassette express a protein that is both recognized by anti-gp120 monoclonal antibodies (mAbs) and is the appropriate size for nonglycosylated full-length gp120 (52 kDa). Immunoblot analysis also demonstrates that the recombinant S. typhi strains express the rgp120 as monomers and multimers found predominantly in the insoluble fraction of the bacteria. Antigen-capture ELISA, using antibodies specific for continuous epitopes on gp120, revealed that the exposure of these epitopes on S. typhi-expressed rgp120 differs from exposure of these epitopes on baculovirus-expressed rgp120 that binds CD4. Epitopes in the first conserved region (109-113) and the third conserved/fourth variable regions (376-380, 382-384, 395-400) are more "surface exposed", while one epitope in the third variable region (313-324) is more "buried" relative to the corresponding epitopes of baculovirus expressed gp120. Antibodies recognizing discontinuous epitopes of the CD4 binding domain do not react with the S. typhi expressed rgp120.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7483776 TI - Anti-HBV neonatal immunization with recombinant vaccine. Part II. Molecular basis of the impaired alloreactivity. AB - HLA study was performed in 9 absolute non-responder (serum titre of anti-HBsAg < 2 mIU ml-1) and 8 hyporesponder (serum antibody level between 2 and 9.9 mIU ml-1) babies who underwent, in neonatal period, HBV vaccination with Engerix B recombinant vaccine. The investigation pointed out that many of these subjects carry HLA haplotypes classically involved in autoimmune diseases: namely HLADR7; DQ2, DR4; DQ8 and DR3; DQ2. The genomic typing for DRB1, DRB3, DRB4, DRB5, DQA1, DQB1 and DPB1 genes revealed an increased frequency of the DRB1*0701; DQA1*0201; DQB1*0201 haplotype (23.5 vs 9.9% of the controls) and of DPB*0201 allele (42.3 vs 13.2% of controls). The polymorphism of Bf, C4A and C4B complement serum components, recognized as important "immune-function-related genes", pointed out an increased frequency of the null allele C4AQ0 (34.3 vs 6.8% of the controls) stressing the role of C4A serum complement component in response to foreign peptide. The immunogenetic investigation has been extended to 23 responder babies (titre of anti-HBsAg > 50 mIU ml-1), vaccinated with the same trial as the poor responders. The HLA frequencies observed in this group were comparable to those of control population and, with respect to the HLA markers cited above, absolutely different from the non/hyporesponder infants. From the HLA class II sequence analysis in the group of poor-responder babies some characteristics, peculiar to autoimmune diseases, have been observed: the majority of the infants showed at least an arginine at the 52 residue of the alpha chain of DQ molecule and a non-aspartic acid at the 57 position of the DQ beta chain.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7483779 TI - Two novel T cell epitope prediction algorithms based on MHC-binding motifs; comparison of predicted and published epitopes from Mycobacterium tuberculosis and HIV protein sequences. AB - We have designed two computer-based algorithms for T cell epitope prediction, OptiMer and EpiMer, which incorporate current knowledge of MHC-binding motifs. OptiMer locates amphipathic segments of protein antigens with a high density of MHC-binding motifs. EpiMer identifies peptides with a high density of MHC-binding motifs alone. These algorithms exploit the striking tendency for MHC-binding motifs to cluster within short segments of each protein. Putative epitopes predicted by these algorithms contain motifs corresponding to many different MHC alleles, and may contain both class I and class II motifs, features thought to be ideal for the peptide components of synthetic subunit vaccines. In this study, we describe the use of OptiMer and EpiMer for the prediction of putative T cell epitopes from Mycobacterium tuberculosis and human immunodeficiency virus protein antigens, and demonstrate that these two algorithms may provide sensitive and efficient means for the prediction of promiscuous T cell epitopes that may be critical to the development of vaccines against these and other pathogens. PMID- 7483780 TI - Efficacy study of a new albumin-free human diploid cell rabies vaccine (Lyssavac HDC, Berna) in 100 severely rabies-exposed Thai patients. AB - A newly developed human diploid cell rabies vaccine (Lyssavac-HDC), produced without added serum albumin and with an effort to remove the virus-inactivating beta-propriolactone prior to addition of the gelatin, L-cysteine and potassium phosphate stabilizer, was tested for safety immunogenicity, adverse reactions and efficacy in 100 severely rabies-exposed Thais. All patients also received human rabies immune globulin and vaccine was administered using the conventional 5-dose intramuscular schedule of one dose on days 0, 3, 7, 14 and 28. One hundred percent of a subgroup of 40 subjects, where blood had been collected, had neutralizing antibodies greater than 0.5 IU ml-1 on days 28 and 90 and all had detectable titers on days 7, 14, 28, 90, 180 and 360. All patients could be followed for at least 1 year and remained well. No significant side-effects from this vaccine were noted. PMID- 7483781 TI - Development of an alternative method for testing the immunogenicity of diphtheria vaccines. AB - The immunogenicity of the diphtheria component of 73 commercial vaccines from five different manufacturers was tested by the toxin neutralization test (TNT) and the enzyme-linked immunosorbent assay (ELISA) developed in our laboratory. A comparison of the antibody levels measured by both assays showed a very good correlation (r = 0.95, p < 0.001). The results suggest that the proposed ELISA is a reliable, simple and economical alternative to the TNT in guinea pigs. Also, the ELISA was found to measure IgG antibody levels as low as 5.5 x 10(-5) IU ml 1. To evaluate the possibility of accelerating the active immunization during the activity test of vaccines, an alternative schedule using one single human dose was assayed. A very good correlation was observed between the IgG antibody response obtained with this schedule and with the traditional programme. Therefore, the cost and the time required to perform the activity test may be considerably reduced when both the rapid immunization schedule and the ELISA are used. PMID- 7483782 TI - Immunization of mice with recombinant Sjc26GST induces a pronounced anti fecundity effect after experimental infection with Chinese Schistosoma japonicum. AB - We report the cloning, by polymerase chain reaction (PCR), of a cDNA encoding a Schistosoma japonicum (Chinese) 26 kDa glutathione-S-transferase (GST) (Sjc26GST), expression of the cDNA, affinity purification of the recombinant GST and its vaccine efficacy in outbred NIH mice using Freund's as adjuvant. The most striking feature of the vaccination experiments was the pronounced reduction in the number of eggs in the livers and spleens of immunized mice. A relatively low but significant level of protection in terms of reduced worm viability against challenge infection was also observed. Further, the level of anti-Sjc26GST antibody in immunized mice was significantly higher than in control mice at week 6 post-challenge infection. These results closely mirror the protection conferred by immunization of animals with the 28 kDa GST of S. mansoni (Sm28) where a reduction in worm viability, worm fecundity and egg-hatching ability have been reported following challenge with S. mansoni. In terms of developing a vaccine against schistosomiasis japonica, immunization with Sjc26GST can provide two complementary goals in human or animal populations--some reduction in worm burden following exposure to infection or reinfection, and an anti-disease effect through reduction of pathology by a decrease in worm fecundity, with this direct effect also affecting the transmission of S. japonicum. PMID- 7483784 TI - Cytotoxic T lymphocyte memory: role in cross-protective immunity against influenza? PMID- 7483785 TI - Setting priorities for the Children's Vaccine Initiative: a cost-effectiveness approach. AB - To help the Children's Vaccine Initiative (CVI) achieve its goal of new and improved children's vaccines, we developed and applied a cost-effectiveness model to set priorities for vaccine development. The model measures the health benefits in additional Quality-Adjusted Life Years (QALYs) gained by the combined birth cohorts of all developing countries over an assumed useful life of a proposed vaccine (generally 10 years). It measures costs as the net cost of developing, procuring, and administering the vaccine to the same population and time frame compared to the status quo (the current vaccine, if any). It weights each dollar of in-kind allocation of the existing health infrastructure less heavily than a dollar cash outlay to purchase new vaccine to reflect severe constraints on foreign exchange and non-personnel costs. It expresses cost-effectiveness as the net cost per QALY. The model was applied to 13 candidate vaccines selected by the CVI for initial analysis on the basis of their near-term feasibility. The five most cost-effective improvements, each of which could generate a QALY inexpensively (below $25 per QALY), were an early-administration or an early two dose measles vaccine, slow release tetanus toxoid (for women), improved typhoid vaccine, and hepatitis B combined with diphtheria-tetanus-pertussis vaccine. PMID- 7483783 TI - Molecular epidemiology of hepatitis B virus vaccine variants in Singapore. AB - Perinatal infection with variants of hepatitis B virus occurs despite combined immunoprophylaxis with hepatitis B immunoglobulin and currently licensed plasma derived and recombinant yeast hepatitis B vaccines. Several variants have been detected during a large study of infants born to carrier mothers in Singapore. The most frequent variant was a virus in which a single amino acid substitution Gly to Arg occurred at amino acid position 145 of the outer protein coat of the virus. Similar mutations have been described in Italy, Japan, the USA and a number of other countries. The emergence of antibody escape mutants is a cause for concern for the detection of virus and possibly for future immunization programmes. PMID- 7483787 TI - Antibody responses by cattle after vaccination with commercial viral vaccines containing bovine herpesvirus-1, bovine viral diarrhea virus, parainfluenza-3 virus, and bovine respiratory syncytial virus immunogens and subsequent revaccination at day 140. AB - Calves were vaccinated with four different commercial viral vaccines containing bovine herpesvirus-1 (BHV-1), bovine viral diarrhea (BVDV), parainfluenza-3 virus (PI-3V), and bovine respiratory syncytial virus (BRSV) immunogens. For the initial vaccination certain vaccines were given twice (days 0 and 28), whereas other vaccines were given on day 0. The calves received another injection on day 140 with the vaccine originally given on day 0. The sera were collected at days 0, 7, 14, 21, 28, 42, 56, 84, 112, 140, 154, 168, and 196 and assayed for viral neutralizing antibodies. The calves were seronegative to BHV-1, BVDV, and BRSV at the onset of the experiment; however, the calves were PI-3V antibody positive due to prior active infection. The commercial vaccines were: (I) inactivated; (II) modified live virus (MLV); (III) combination of chemically altered live virus, MLV, and inactivated virus; and (IV) combination of inactivated and MLV. Among the vaccine groups there were differences in onset and duration of antibodies as measured by geometric mean titers to each immunogen in postvaccination collection dates compared to day 0 titers; and likewise compared to day 140 titers after revaccination at day 140. There were also differences in antibody titers to the various viruses among the vaccine groups on specific collection dates. All four vaccines induced increased BHV-1 antibodies by day 14 after the initial injection. The antibody titers induced by MLV BHV-1 and the chemically altered BHV-1 vaccines had greater duration than those induced by the inactivated vaccine.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7483786 TI - Efficacy trials with tissue-culture grown, inactivated vaccines against chlamydial abortion in sheep. AB - Vaccines containing inactivated, semi-purified antigen of ovine abortifacient Chlamydia psittaci were prepared from tissue-culture grown harvests of two strains. These were evaluated for efficacy against experimentally reproduced enzootic abortion of ewes (EAE), the protection index (PI) of each vaccine being calculated from three variables. Trial 1 tested the adjuvants Marcol 52/Arlacel A, ISCOM matrix, Novasomes and Alhydrogel, each administered with a total antigen dose of 4 micrograms chlamydial protein (cp), against challenge with the two homologous vaccine strains. The first three formulations yielded PIs of 39-42%, while that of Alhydrogel was 23%. Marked granuloma and/or abscess formation occurred in the Marcol 52/Arlacel A group but in none of the others, and only this group seroconverted following vaccination. In Trial 2, ISCOM matrix containing 16 micrograms cp per dose was administered once or twice. Two challenges were used, one involving the two vaccine strains and the other these two strains plus five others, but no significant differences were seen between them. The double dose schedule, which was better than the single dose and also several Novasomes formulations tested concurrently, gave PIs of 64 and 77% against the two challenges (combined value 71.5%). Trial 3 studied duration of immunity over two lambing seasons using three different administration schedules of 16 micrograms cp in ISCOM matrix, namely single dose in year 1, single dose in year 2 (Group 1), double dose then booster (Group 2), and double dose without booster (Group 3). PIs against the homologous bivalent challenge were 79-80% for Groups 1 and 3 and 91% for Group 2.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7483788 TI - A molecular and serologic study of the envelope gene of the British isolate: HIV 1 GB8. AB - The entire envelope gene of a British HIV-1 isolate, GB8, was cloned, sequenced and aligned with those of the reference strains MN, SF2 and IIIB/LAI. Three of the viruses (MN, IIIB/LAI, GB8) and their recombinant gp120s, were then characterised using an extensive panel of human HIV-1 positive sera and mapped neutralising monoclonal antibodies (MAbs). Overall, the GB8 env-gene translation product shares 84% homology with those of the reference strains. Across the V3 region homology was greater between GB8 and SF2/MN (74.3-88.6%) than IIIB/LAI (63.9-66.7%). Accordingly, GB8 was sensitive to V3-specific MAbs which neutralise MN/SF2 and resistant to those that neutralize IIIB/LAI. In the CD4 binding region the central MWQEVGKAMYAPPI was conserved in all viruses but homology in the N terminus was greater between GB8 and SF2 and IIIB/LAI than MN. GB8 and IIIB/LAI were sensitive to all MAbs raised against the CD4 binding site whereas MN was resistant to 3 of 4 tested. Human sera obtained from a London-based cohort did not differentiate between GB8 and MN in neutralisation assays, whereas IIIB/LAI titres were significantly lower at all stages of disease. These results show that GB8 carries a consensus-like V3 loop and is as representative as MN of HIV-1 viruses circulating in the UK. To our knowledge, GB8 is the only British HIV-1 isolate which has been characterised to date. PMID- 7483789 TI - Rapid vaccination protocols for commercial vaccines against tick-borne encephalitis. AB - Although inactivated viral vaccines have been dramatically successful in controlling many of the world's most devastating diseases, they frequently need several injections to ensure high levels of protection, and thus their efficacy is reduced in many situations. We have developed several rapid vaccination protocols for two commercial vaccine preparations against tick-borne encephalitis virus and studied their efficacy in an experimental murine model. Vaccination protocols as brief as two doses given over two days elicit efficient protection against challenge with potentially fatal doses of virus and this protection is afforded as soon as 5 or as long as 100 days after the first vaccination. The very rapid induction of protection and the poor antibody responses observed would suggest that cell-mediated immune responses are the most important mechanisms for the protection elicited by conventional inactivated vaccines against tick-borne encephalitis. PMID- 7483790 TI - The purification and protective capacity of Bordetella pertussis outer membrane proteins. AB - The whole cell vaccine (WCV) of Bordetella pertussis is protective in the intracerebral (i.c.) mouse protection assay. We found a correlation between the i.c. mouse protection assay potency and the presence of the virulence-associated outer membrane proteins (OMPs) in outer membrane complexes (OMC). The virulence associated 92, 32 and 30 kDa OMPs were purified and the N-terminal amino acid sequences were determined. The N-terminal amino acid sequences of the 30 and 32 kDa OMPs show homology with the C-terminal fragment of the P.93 precursor of the 69 kDa OMP (pertactin). The purified 32 kDa OMP was protective in the i.c. test when presented as mixed protein-detergent micelles. The 92 kDa OMP became a protective antigen when nonprotective levels of pertussis toxin were added. We found a correlation between the i.c. mouse protection value and the 92 kDal38 kDa (porin) ratio in OMC preparations. PMID- 7483791 TI - P-fimbriae of Escherichia coli as carriers for gonadotropin releasing hormone: development of a recombinant contraceptive vaccine. AB - The demand for an effective and low cost means of fertility control of domestic animals has raised interest in the development of contraceptive vaccines. A promising candidate for a vaccine component is the brain peptide gonadotropin releasing hormone (GnRH), which plays a central role in the regulation of reproductive functions in vertebrates. Neutralization of GnRH by vaccine-induced antibodies is expected to prevent the reproductive activity in a wide range of species. A GnRH-protein conjugate was prepared by means of recombinant DNA technology. The oligonucleotides encoding GnRH were inserted in hypervariable regions of the subunit gene of P-fimbriae of Escherichia coli. Hybrid fimbriae encoded by the manipulated fimbrial gene clusters were expressed efficiently on the cell surface of Escherichia coli. Vaccination of female rats and bull calves with purified hybrid GnRH-fimbriae strongly affected the reproductive characteristics. We conclude that P-fimbriae represent a very attractive carrier system for application of GnRH in a new type of vaccine. PMID- 7483792 TI - Vaccination against tick-borne encephalitis (TBE): influence of simultaneous application of TBE immunoglobulin on seroconversion and rate of adverse events. AB - One hundred and twenty-one of 128 enrolled healthy adult subjects were immunized against tick-borne encephalitis (TBE) either by TBE vaccine on days 0 and 28 (61 subjects) or simultaneously by TBE vaccine plus TBE immunoglobulin on day 0 plus TBE vaccine on day 28 (60 subjects). Formation of TBE antibodies were measured in ELISA on days 0, 28 and 56. On day 28 median TBE antibodies were twice as high in the vaccine group (2400) as in the vaccine plus immunoglobulin group (1200). Adverse events were more often observed after the first vaccination than after the second in both groups. In the vaccine group, adverse events after the first vaccine dose were remarkably more frequent (45% of these subjects reported 38 adverse events) than in the group who received vaccine plus immunoglobulin (25% of these subjects reported 18 adverse events). All types of adverse events (chills, flu-like symptoms, injection site pain) were reported less frequently in the vaccine plus immunoglobulin group. After the second vaccination the rate of adverse events was 7% in both groups. Seroconversion was achieved in all subjects on day 56 except one subject in the vaccine plus immunoglobulin group. Simultaneous application of TBE vaccine plus TBE immunoglobulin can be recommended for persons who need immediate protection plus active TBE vaccination. PMID- 7483793 TI - Immunogenic and protective properties of chicken anaemia virus proteins expressed by baculovirus. AB - The coding information for three putative chicken anaemia virus proteins (VP1, VP2, VP3) was inserted into a baculovirus vector and expressed in insect cells. The immunogenic properties of the chicken anaemia virus (CAV) proteins produced separately or together in insect-cell cultures were analysed by inoculating them into chickens. Only lysates of insect cells which have synthesised equivalent amounts of all three recombinant CAV proteins or cells which synthesised mainly VP1 plus VP2 induced neutralising antibodies directed against CAV in inoculated chickens. Progeny of those chickens were protected against clinical disease after CAV challenge. Inoculation of a mixture of lysates of cells that were separately infected with VP1-, VP2- and VP3-recombinant baculovirus did not induce significant levels of neutralising antibody directed against CAV and their progeny were not protected against CAV challenge. Our results indicate that expression in the same cell of at least two CAV proteins, VP1 plus VP2, is required to obtain sufficient protection in chickens. Therefore, recombinant CAV proteins produced by baculovirus vectors can be used as a sub-unit vaccine against CAV infections. PMID- 7483794 TI - Immunity to diphtheria in the 4-70 year age group in Italy. AB - Diphtheria antibody level in serum samples obtained from 270 subjects aged 4-70 was measured by in vitro seroneutralization test on Vero cells. Of the studied population, 27.8% had an antibody titre below the protective level (< 0.01 IU/ml). The prevalence of susceptible subjects showed a significant age-related increase (p < 0.01), with the highest value (53.8%) in the 31-40 age group. Mean antibody titre was maximum in individuals aged 13-14, then decreased reaching the minimum level in the 41-50 age group. These data suggest that in individuals aged 30-50, diphtheria immunity is not satisfactory, both for prevalence of immune subjects and for antibody levels. Therefore, a revaccination of adults with reduced doses of diphtheria toxoid may be advisable. PMID- 7483796 TI - Antigenic variants in a plaque-isolate of foot-and-mouth disease virus: implications for vaccine production. AB - The occurrence of many subtypes within a serotype of foot-and-mouth disease virus (FMDV) makes it difficult to control the disease by vaccination. Although inactivated vaccines are used successfully in many countries, the appearance in the field of antigenic variants against which the vaccines do not confer protection is a constant problem in vaccine manufacture. We had found previously a mixture of antigenic variants in a field isolate of serotype A12. In this report we demonstrate the presence of two variants in a plaque-isolate from this mixture. The second variant was detected only when the growth conditions were altered. Our observation points to the problems which may be encountered in the large scale growth of a virus for vaccine production. PMID- 7483797 TI - Measles vaccination in India and controversies regarding adverse reactions. PMID- 7483795 TI - Immunization of mice by oral colonization with live recombinant commensal streptococci. AB - To test the use of recombinant streptococci as live vaccine vectors, colonization/immunization experiments were performed with Streptococcus gordonii expressing heterologous cell-surface antigens. Three isogenic strains of S. gordonii were used: a wild-type, a recombinant expressing the M6 protein of Streptococcus pyogenes, and a recombinant expressing the E7 protein of human papillomavirus type 16 as a fusion with the M6 protein. A single dose of live bacteria was used to inoculate outbred mice, and it was found that: (i) mice were stably colonized by a single intranasal/oral inoculum of S. gordonii; (ii) recombinant strains were equally effective as wild-type in colonizing mice; (iii) two months after the inoculum, oral/pharyngeal swabs of 83.3% of animals were still positive for isolation of S. gordonii; (iv) recombinant S. gordonii isolated from colonized mice were always positive for expression of the heterologous antigens; (v) live bacteria induced a systemic immune response, since sera of mice colonized with recombinant S. gordonii contained IgG specific for the heterologous cell-surface antigens; (vi) this immune response depended upon the effective colonization by live bacteria, since killed bacteria did not induce such a response. PMID- 7483798 TI - Comparison of three different schedules of vaccination against hepatitis B in health care workers. AB - Health care workers (HCW) are a group at risk for hepatitis B virus (HBV) infection; as a result, vaccination is recommended. However, elevated cost of the vaccination schedule is one of the limiting factors to this approach. Our aim in this study was to evaluate alternative schedules for vaccination against hepatitis B, in order to obtain safe immunization with reduced costs. We studied 300 HCW, randomized to be submitted to one of three vaccination schedules against hepatitis B: Group A--three doses of 20 micrograms i.m. (n = 103); Group B--first dose i.d. (2 micrograms), second and third doses i.m. (20 micrograms) (n = 97); Group C--first and second doses i.d. (2 micrograms), third dose i.m. (20 micrograms) (n = 100). All individuals received recombinant vaccine at 0, 1 and 6 months. After the first dose, there was no difference among the three schedules, either in terms of anamnestic response or in seroconversion rate. After the second dose, there was statistical difference among the three schedules (A > B > C), in terms of seroconversion rates. After the third dose, seroconversion rates were 92.2% in Group A and 92.8% in group B; geometric mean titers (GMT) in Group B (789.6 UI l-1) were similar to group A (1248.0 UI l-1). Group C presented a seroconversion rate of 78% and a GMT of 323.0 UI l-1, both statistically inferior to other schedules. We concluded that the schedule applied in Group B had similar results when compared to schedule A, with estimated savings of 30% in vaccine costs.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7483799 TI - Effective antibody response in newborn babies living in Maldives to simultaneous vaccination against hepatitis B, poliomyelitis, diphtheria and tetanus. AB - The antibody responses of Maldivian infants early in their life to simultaneous immunization against hepatitis B virus, poliomyelitis, diphtheria and tetanus were investigated. The vaccines were given at 6, 10 and 14 weeks of age. Among 243 newborn babies from HBsAg-negative mothers, 103 received three doses of oral poliomyelitis (OPV) and diphtheria and tetanus (DTV) vaccines; 105 were similarly immunized but received in addition the recombinant hepatitis B vaccine (HBV); 35 were immunized with the HBV recombinant vaccine alone. The antibody response to all of the vaccines was effective. No significant differences among the groups were observed. Hepatitis B vaccination of infants neither affected nor was affected by the contemporary administration of OPV and DTV vaccines. PMID- 7483800 TI - Antibodies to measles, mumps and rubella in UK children 4 years after vaccination with different MMR vaccines. AB - Persistence of antibodies 4 years after vaccination with measles, mumps and rubella (MMR) vaccine from three different manufacturers was compared in 475 children who received a single injection of vaccine when aged 12-18 months. Antibodies to measles and mumps were measured using a plaque reduction neutralisation assay; rubella antibodies were measured by radial haemolysis and latex agglutination. Children given MMR vaccine containing the Urabe mumps strain were less likely to be antibody negative than those given the Jeryl Lynn mumps strain (39/266, 15% vs 39/204, 19%, p = 0.048). However, the relatively high proportions in both groups without detectable mumps neutralising antibody suggests the probable need for a second dose in order to achieve mumps elimination. No significant differences were found in the proportions with detectable antibody to measles between vaccines containing the Schwarz and the Enders-Edmonston strains. Overall, only 3% of vaccinees were without detectable measles antibody, although a further 28% had a level below 200 mille International Units, the putative protective level for clinical measles. Geometric mean titres (GMTs) to measles were twofold higher in girls vaccinated after than before 14 months of age; GMTs in boys were intermediate and showed no age effect. Over 99% of vaccinees were seropositive to rubella, confirming the excellent immunogenicity of the RA 27/3 rubella strain and the potential for elimination of rubella with a single dose strategy. PMID- 7483801 TI - Immunoregulatory role of H-2 and intra-H-2 alleles on antibody responses to recombinant preparations of B-subunits of Escherichia coli heat-labile enterotoxin (rEtxB) and cholera toxin (rCtxB). AB - The immunoregulatory role of H-2 and intra-H-2 alleles on antibody responses to recombinant preparations of B-subunits of Escherichia coli heat-labile enterotoxin (rEtxB) and cholera toxin (rCtxB) is reported. Oral delivery of rEtxB to congenic mice of several different H-2 haplotypes resulted in H-2 dependent serum IgG responses (H-2d > H-2b = H-2q > H-2a > H-2k) and a similar spectrum of intestinal IgA responses in those strains tested. Responses to rEtxB and rCtxB were found to be differentially modulated by the H-2 locus, with significant differential effects in H-2b and H-2d congenic strains (H-2d > H-2b for rEtxB; H 2b > H-2d for rCtxB). Additionally, it was found that when rEtxB was fed to mice previously primed (orally) with either rEtxB or rCtxB only those mice primed with rEtxB exhibited a booster response. A second booster immunisation with rEtxB in rCtxB-primed mice produced an H-2 dependent spectrum of responses characteristic of those elicited by rEtxB, with the antibodies predominantly directed against rEtxB and not rCtxB. These results indicate that the differential response to rEtxB and rCtxB is set at the T- and B-cell level. Also, immunoregulation of antibody responses to rEtxB by intra-H-2 I-E in mice transgenic for the entire IEka gene was investigated. No significant difference between responses in transgene-positive and -negative mice was found, suggesting that antigen presentation does not involve I-E, but occurs in the context of I-A.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7483802 TI - Circumstances and management of 72 animal bites among long-term residents in the tropics. AB - Little is known about rabies exposure among expatriates living in warm climates. Recommendations on pre-exposure prophylaxis are therefore controversial. This study assesses the post-exposure management of Swiss and German expatriates after potential rabies exposures. Dogs were involved in 69% of all incidents. Less than half of the owned dogs were vaccinated against rabies. Only 24-30% of post exposure treatments were correct according to WHO recommendations. Expatriates with pre-exposure vaccination appear to be complacent about post-exposure treatment. The decision to give pre-exposure vaccination to expatriates in warm climate countries or to rely on post-exposure rabies vaccination depends on the available quality of treatment and must be carefully assessed prior to a stay abroad. Information on rabies risk must be better disseminated and vaccination of owned animals improved. PMID- 7483803 TI - A method to screen T lymphocyte epitopes after oral immunisation of humans: application to cholera toxin B subunit. AB - The response to oral immunisation of humans with classical biotype cholera toxin B subunit was studied to identify immunodominant T lymphocyte determinants. The in vitro proliferative response to pools of 12-mer peptides and larger peptides used individually was analysed by a novel statistical approach, and identified an immunodominant region in residues 70-79 in immunised subjects, when either pools or individual peptides were employed. In contrast, a patient infected with El Tor biotype had a dominant response to residues 40-60. The statistical software employed in this study may enable efficient screening of antigens for immunodominant T lymphocyte determinants when blood precursor frequencies are low following immunisation, and may therefore be of special relevance to mucosal vaccines. PMID- 7483804 TI - Efficacy, safety, and immunogenicity of a meningococcal group B (15:P1.3) outer membrane protein vaccine in Iquique, Chile. Chilean National Committee for Meningococcal Disease. AB - A meningococcal group B (15:P1.3) outer membrane protein vaccine was tested for efficacy in a randomized, double-blind controlled study in Iquique, Chile. A total of 40 811 volunteers, ages 1-21 years, enrolled in the study. Volunteers received two doses of vaccine six weeks apart by jet injector. Both the experimental vaccine and the control vaccine (Menomune, A, C, Y and W135 meningococcal polysaccharide vaccine) were well tolerated with minor side effects. Active surveillance for suspected cases of meningococcal disease was conducted for 20 months in Iquique. Eighteen cases of group B meningococcal disease were confirmed during the 20 months. Efficacy was estimated to be 51% (p = 0.11) for all ages combined. In children aged 1-4 no protection was evident, but in volunteers aged 5-21 vaccine efficacy was 70% (p = 0.045). The IgG antibody response by ELISA was characterized by a large booster effect after the second dose, followed by a substantial drop in antibody levels by 6 months. The youngest children had the highest responses. The bactericidal antibody response, on the other hand, was characterized by the lack of a significant booster response, higher responses in the older children, and an increase in the geometric mean titer in the later months of the study in the older children. PMID- 7483805 TI - Immunogenicity of the yeast recombinant p17/p24:Ty virus-like particles (p24-VLP) in healthy volunteers. AB - In this Phase I study, immunisation with the yeast-derived p24 virus-like particles Ty p24-VLP (3 x 100 or 3 x 500 micrograms subcutaneously) in 16 healthy male subjects elicited p24 antibody responses in 4 of 16 (25%) subjects. After a fourth, intramuscular, immunisation (500 micrograms), p24 antibody responses were detected in 11 of 15 (70%) subjects. In addition to p24 antibody responses, T cell proliferative responses were also observed, although no HLA restricted p24 specific cytotoxic T cell responses were detected. The results demonstrate that Ty p24-VLP is immunogenic and well-tolerated in healthy male subjects. PMID- 7483806 TI - Antigenicity of hepatitis A virus after ultra-violet inactivation. AB - Ultra-violet (UV) treatment has been shown to inactivate hepatitis A virus (HAV) in wastewater and polluted drinking water. Whether this method could be used to inactivate virus preparations made for vaccine purposes is not known since the effect of UV on the antigenicity of HAV has not been studied. HAV vaccine preparations have been treated effectively with formaldehyde. However, this method is time-consuming, since treatment times of up to 15 days have been published as necessary for a complete and safe inactivation. We used a cell culture-derived HAV preparation with a TCID50 of 10(9) for a UV irradiation experiment. The antigenicity (assessed by a panel of anti-HAV antibodies), viral genome titre (quantitated by polymerase chain reaction) and HAV infectivity were compared after treatment with UV doses of 0, 184, 368, 552, 736 and 920 J m-2. Our results showed the antigenicity of HAV was almost unaltered even when infectious viral particles were no longer detectable. This technique shows potential as a simple and low-cost method for an inactivated HAV vaccine. PMID- 7483807 TI - Immunogenicity of influenza A virus N2 neuraminidase produced in insect larvae by baculovirus recombinants. AB - Influenza A virus neuraminidase (NA) from A/Udorn/72 (H3N2) was expressed by recombinant baculovirus-infected insects. The recombinant NA was enzymatically active. Enzyme activity was neutralized by polyclonal antisera raised against virion-extracted NA. NA produced in whole insects by a baculovirus expression system is antigenically indistinguishable from virion NA by polyclonal antisera in functional assays (NI) and in ELISA, and is highly immunogenic without adjuvant. It is equivalent in immunogenicity to NA purified from influenza virus. Our results indicate that baculovirus-produced NA could be used as a source for large quantities of purified N2-NA for vaccine use. PMID- 7483808 TI - Cold-passaged, temperature-sensitive mutants of human respiratory syncytial virus (RSV) are highly attenuated, immunogenic, and protective in seronegative chimpanzees, even when RSV antibodies are infused shortly before immunization. AB - A cold-passaged (cp) temperature-sensitive (ts) RSV mutant, designated RSV cpts 530, which possesses host-range mutations acquired during 52 passages at low temperature in bovine tissue culture and one or more ts mutations induced by chemical mutagenesis (shut-off temperature 39 degrees C) was found previously to be tenfold restricted in its replication in mice as compared to wild-type virus and stable genetically in nude mice. In the current study, we introduced additional attenuating mutations, such as small-plaque (sp) or ts mutations, into cpts-530 by chemical mutagenesis with 5-fluorouracil, with the intent of obtaining derivatives of cpts-530 that were more attenuated in mice or chimpanzees and that were more stable genetically following replication in vivo. Fourteen mutants of RSV cpts-530 which had acquired an additional ts mutation were identified and found to be more restricted in replication in BALB/c mice than the cpts-530 parental strain. One mutant, designated cpts-530/1009 (shut-off temperature 36 degrees C), was 30 times more restricted in replication in the nasal turbinates of mice and threefold more restricted in the nasopharynx of seronegative chimpanzees than its cpts-530 parent. Like its parent, this mutant was highly restricted (30,000-fold) in replication in the lower respiratory tract of chimpanzees even following direct intratracheal inoculation. The cpts-530 and cpts-530/1009 mutants exhibited a high level of stability of the ts phenotype during replication in chimpanzees.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7483809 TI - Priming of class I-restricted cytotoxic T lymphocytes by vaccination with recombinant protein antigens. AB - We investigated the specific priming of MHC class I-restricted cytotoxic T lymphocytes (CTL) by different protein antigen preparations in mice. The recombinant viral protein antigens tested are of potential relevance for the design of subunit vaccines. They include the hepatitis B virus (HBV) surface antigen (S-antigen), the HIV-1 gp160 envelope protein, and a chimeric HIV-1 Pr55 gag/V3-3 retrovirus-like particle. In addition, ovalbumin (OVA) was tested. The native or denatured particulate (multimeric) or monomeric form of these protein antigens was injected by various routes into mice. Class I-restricted CTL were efficiently primed by a single low-dose injection of HBV S-antigen particles or the chimeric HIV-1 Pr55-gag/V3-3 particles. After SDS-denaturation, gel-purified monomeric S-antigen and monomeric Pr55-gag/V3-3 fusion protein were still very efficient in priming CTL. CTL sensitization was not detected in a (primary or boosted) response to even high doses of native OVA or native HIV-1 gp160. Denaturation of these two antigens by detergent strikingly increased their immunogenicity for CTL. Immunization of mice with non-treated or SDS-denatured antigenic peptides representing the relevant CTL-defined epitopes of the tested protein antigens did not prime CTL. These data indicate that native, particulate and denatured, monomeric protein antigens efficiently stimulate a class I restricted CTL response. PMID- 7483810 TI - Vaccination with whole-cell vaccine and bacterial protein extract protects tilapia against Streptococcus difficile meningoencephalitis. AB - Formalin-killed Streptococcus difficile strains used as vaccines delivered intraperitoneally were able to protect tilapia against a challenge of 100 LD50. The protection obtained was not strain specific. A vaccine based on an S. difficile extract containing 50% protein conjugated to alum also protected tilapia challenged with a virulent S. difficile strain. Protection in tilapia was correlated with the development of specific agglutinins. Western blot analysis supported the hypothesis that only a few proteins act as protective antigens in both the whole-cell vaccine and the streptococcal extract. The high efficacy of these vaccines make them good candidates for the control of streptococcal fish meningoencephalitis. PMID- 7483811 TI - BHV-1 glycoprotein 1 and recombinant interleukin 1 beta efficiently elicit mucosal IgA response. AB - The mucosal immune response to most soluble antigens administered directly to the mucosal system is low and requires a large amount of antigen and frequent vaccinations. In this study we tested whether immunizing cattle at a site which shares lymphatic drainage with the nasal mucosa could prime local mucosal immunity. We further tested whether recombinant bovine IL-1 beta (rBoIL-1 beta) could potentiate the induction of mucosal immunity. Animals were immunized subcutaneously at the base of the ear (s.e.) with recombinant bovine herpesvirus 1 (BHV-1) envelope glycoprotein I (gI) (35 micrograms animal-1) emulsified in incomplete Freund's adjuvant with or without rBoIL-1 beta (500 ng kg-1) followed by a second immunization 42 days later. Animals were challenged with virulent BHV 1 intranasally 42 days after the second immunization. Mucosal IgA from the nares was induced after only one immunization, and enhanced by boosting. rBoIL-1 beta treated animals had higher levels of BHV-1 specific nasal IgA (p < 0.01) and serum neutralizing antibody (p < 0.05). rBoIL-1 beta-treated animals also had increased numbers of surface IgA+ (p < 0.05) and IgG1+ (p < 0.001) B cells after in vitro antigen (gI) stimulation of peripheral blood lymphocytes suggesting that there was a greater expension of IgA+ and IgG1+ B cells in rBoIL-1 beta treated animals. When challenged with BHV-1, 3 of 4 animals in the gI+rBoIL-1 beta group were fully protected from viral replication in the nares, while only 1 of 4 animals receiving gI alone was protected.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7483812 TI - Astounding influence of 'substance abuse', malnutrition and immune suppression on immune response to hepatitis B vaccine. PMID- 7483813 TI - [Ethical assessment of biomedical experiments in The Netherlands]. AB - The ethical evaluation of biomedical experiments is loyally accepted in the Netherlands. It was introduced everywhere during the last 15 years. Among the targets we find not only the protection of the experimental objects--animals and humans--but also the protection of researchers who may find moral support in the approval of their projects by a Committee of Ethical Evaluation. Concerning experiments in animals, the legal framework is provided by the Law on Animal Experiments. This law was drawn up in accordance with the directive of the E.U. To be allowed to practice experiments in animals, an institute must possess a licence; the researchers are moreover obliged to be appropriately trained with regard to experiments in animals. To that end an adequately functioning organization has been set up. The intensive supervision is widely decentralized and effected for the greater part by experts working within the institutes. The "intra muros" Committees supervising the experiments in animals play an important part in the ethical evaluation. More than 50 of those Committees are active at this time. Setting the ethical standards is done in close collaboration between the Authorities and the researchers. Researchers and animal protection associations have established a "Platform for the Replacement of Experiments in Animals", in which they support development of alternative methods for research. The legislation concerning medical experiments in humans is not yet completely enforced, but in practice the ethical evaluation has been effected for many years in every hospital of the Netherlands. At present about 150 "Committees for medico ethical evaluation" are at work. Their task may be very heavy, especially in the academic hospitals, where, mostly, over 150 projects are advised on every year. Adequate training facilities are provided for the members of the committees. The passing of the bill on experiments in humans is stagnating owing to political reasons. Besides the approval of the present practice, the bill-draft contains some elements that are difficult to accept by researchers; among other things, the researchers refuse a possible political influence on the ethical advice. Moreover, there exists a menace to see the insurers changing their rules: the legislation on privacy might also hamper the development of a research seeming too closely patient-linked. Sufficient attention is not given to the financial aspects of the evaluation-process. The real cost of the evaluation should be incorporated into the budget of research projects. PMID- 7483814 TI - Staphylokinase: fibrinolytic properties and current experience in patients with occlusive arterial thrombosis. AB - Staphylokinase is a profibrinolytic agent that forms a 1:1 stoichiometric complex with plasminogen which, following conversion to plasmin, activates other plasminogen molecules to plasmin. The plasmin, staphylokinase complex, unlike the plasmin, streptokinase complex, is rapidly inhibited by alpha 2-antiplasmin. In a plasma milieu, staphylokinase is able to dissolve fibrin clots without associated fibrinogen degradation. This fibrin-specificity of staphylokinase is the result of reduced inhibition by alpha 2-antiplasmin of plasmin, staphylokinase complex bound to fibrin, recycling of staphylokinase from the plasmin, staphylokinase complex following inhibition by alpha 2-antiplasmin, and prevention of the conversion of plasminogen, staphylokinase to plasmin, staphylokinase by alpha 2 antiplasmin. In several experimental animal models, staphylokinase appears to be equipotent to streptokinase for the dissolution of whole blood or plasma clots, but significantly more potent for the dissolution of platelet-rich or retracted thrombi. The feasibility of fibrin-specific coronary thrombolysis with an intravenous infusion over 30 min of 10 mg recombinant staphylokinase was demonstrated in two small pilot studies in patients with acute myocardial infarction with angiographically confirmed total occlusion of the infarct-related coronary artery. However, neutralizing antibodies against staphylokinase were demonstrable from the third week on in all patients. Definition of the therapeutic benefit of recombinant staphylokinase will require more detailed dose finding studies followed by randomized efficacy studies against other thrombolytic agents. An interim analysis after 50 patients of a randomized trial of recombinant tissue-type plasminogen activator versus staphylokinase in patients with acute myocardial infarction revealed similar rates of coronary patency at 90 minutes but a significantly higher fibrin specificity of the latter compound. PMID- 7483815 TI - [Obstetrical study of the pelvis: a historical overview]. AB - The female pelvis is fundamental to life itself, but it took ages to grasp this truism. During Antiquity, physicians had no need for the concepts of pelvic dystocia and cephalopelvic disproportion. When Aranzo (16th century), a practising physician, stated that a disproportion between the size of the fetus and that of the bony pelvis was the main culprit of difficult labor, his suggestion was overlooked, so much so that it was Hendrik van Deventer (1701) who came to be coined "father of the pelvic theory". The need for an accurate knowledge of the dimensions of the maternal pelvis and of the fetal skull became imperative in the middle of the 18th century, when forceps extraction had become popular. Levret, but above all William Smellie, made valuable contributions to the pelvic theory, which, however, were weakened by Baudelocque's fallacies concerning the intrinsic value of external pelvimetry. After the first quarter of the 19th century, the epicenter of scientific obstetrics moved to Germany and Austria, and German-speaking practitioners, e.g. Michaelis, Litzmann, and the Naegele, deepened our insight in the mechanism of labor. After the discovery of the X-rays (1895), the use of instrumental pelvimetry declined, but the clinical breakthrough of radiopelvimetry was delayed until the twenties of this century. Radiopelvimetry, a very valuable technique indeed, dispensed a large number of mothers from undergoing abdominal delivery. However, awareness of the hazards of fetal-maternal exposure to ionizing radiation pushed ante- and intrapartum radiopelvimetry from the scene. Thanks to the progress of chemistry, pharmacology and electronics, "dynamic pelvimetry" became the fad, heralded by what is called "active management of labour". The fetus now assumed the role of "dynamic pelvimeter". The historical circle was closed. PMID- 7483816 TI - Immunotherapy with bispecific antibodies. AB - With rare exceptions, monoclonal antibodies as therapeutic agents for cancer, have had limited anti-tumor effects. Attempts to improve the cytotoxicity of antibodies by using them as guiding agents for immune effector cells, have been developed. Numerous in vitro studies have shown that T lymphocytes can be targeted towards any target cell by using bispecific antibodies with specificity of the CD3/TCR complex and a target cell antigen. This approach works extremely efficiently in vitro and justified the hope that bispecific antibodies might be useful for focusing T cells towards tumor cells in vivo. Therefore we have produced bispecific antibodies directed against the membrane expressed idiotype of the murine B cell lymphomas BCL1 and 38C13, and against the murine CD3 complex present on all T lymphocytes. The dual specificity of the hybrid-hybridoma produced bispecific antibodies could be demonstrated by flow cytometry, the induction of T cell proliferation, the induction of IL-2 secretion by polyclonal T cells, and redirected lysis of the relevant target cells. Immunotherapy of tumor bearing animals demonstrated that bispecific antibodies could efficiently target T cells towards the tumor cells, that tumor cell--T cell bridging is established in vivo, and that both T cell subsets contribute to tumor regression resulting in long-term survival and cure of the lymphomas. To streamline the production of bispecific antibodies we recently engineered recombinant bispecific proteins produced by bacteria. Two scFv fragments, one derived from the anti tumor antibody and one derived from the anti-mouse CD3 monoclonal antibody were produced in E. Coli. Both antibody fragments could be easily purified by IMAC chromatography. After in vitro refolding of the recombinant proteins the antigen specificity was retained. Both scFv were expressed as one fusion protein by engineering a 15 amino acid linker between the antibody fragments. The purified bispecific fusion protein behaved in vitro tests as active as the hybrid hybridoma produced bispecific antibody (induction of T cell proliferation and lymphokine secretion, retargeted lysis of tumor cells). Also the fusion protein is able to target T cells towards the tumor cells in vivo, resulting in long term survival of the treated animals. In conclusion, these studies form a sound basis for further investigation of the potential of bispecific antibodies for treatment of patients with cancer. PMID- 7483817 TI - Isolation of caliciviruses from skunks that are antigenically and genotypically related to San Miguel sea lion virus. AB - Caliciviruses were isolated from feces of skunks imported from the north central United States to Canada. Virus isolation was accomplished using adenovirus transformed human kidney (293) cells, swine testes and Vero cells. Plaque size variants were presented, but there was no apparent difference in virus morphology by negative stain or immune electron microscopy. Pigs infected with skunk calicivirus had a slightly elevated body temperature at 3 days postinfection. Although the infected animals seroconverted, no overt clinical signs were observed. Purified infectious genomic skunk calicivirus RNA behaved exactly as San Miguel sea lion virus (SMSV) 1 and 4 genomic RNA in cell culture transfection studies. Of the cell types examined, only primary porcine kidney, 293 and Vero cells supported viral replication. No viral replication was detected in cells of bovine, equine, ovine, caprine or feline origin. The skunk caliciviruses contained a single capsid protein with a relative mobility similar to SMSV virus 1 and 4 capsid proteins. The capsid protein was positive by Western blot analysis with SMSV and vesicular exanthema of swine virus (VESV) antisera. Purified RNA from skunk calicivirus infected cells was subjected to reverse transcription followed by polymerase chain reaction. Nucleotide sequences were identified that had greater than 85% similarity to the 2C and RNA polymerase gene regions of SMSV 1 and 4 and VESV A48. Predicted amino acid sequences of these regions were greater than 95% similar and the partial coding sequence of the polymerase gene contained the YGDD sequence common to positive-strand RNA virus polymerases. PMID- 7483818 TI - A helper T-cell epitope of the E7 protein of human papillomavirus type 16 in BALB/c mice. AB - The helper T-cell response to the E7 protein of human papillomavirus type 16 (HPV16) was studied using BALB/c (H-2d) mice. Twenty-two overlapping synthetic peptides spanning the HPV16 E7 protein were split into 6 groups. Mice were sensitized using mixtures of synthetic peptides corresponding to each of the groups. Lymph node cell suspensions were cultured with the corresponding mixture of synthetic peptides that was used for sensitization. Two mixtures induced a proliferative response. Analysis of the individual peptides from these mixtures showed that two (overlapping) peptides induced a proliferative response. This response was mediated by CD4+ cells. The common region of the two peptides was found to be a single epitope, and a minimal epitope was demonstrated (AHYNIVTFCCK). In conclusion, in contrast to others, we demonstrated a helper T cell response in BALB/c mice. This may be due to the fact that we used synthetic peptides as immunizing agent. The helper T-cell epitopes in HPV16 E7 demonstrated previously are partly overlapping with the (minimal) epitope demonstrated here, underlining the 'public' nature of the epitope. PMID- 7483819 TI - The nucleotide sequence of the fusion protein gene of the peste des petits ruminants virus: the long untranslated region in the 5'-end of the F-protein gene of morbilliviruses seems to be specific to each virus. AB - cDNA corresponding to the fusion protein (F) gene of the vaccine strain of peste des petits ruminants virus (PPRV) was cloned and sequenced. The gene was 2321 nucleotides long excluding the poly(A) tail. As with other morbilliviruses, it had a long G/C rich stretch of about 525 nucleotides. There was no start codon before position 489 in the nucleotide sequence. From the 489th nucleotide to the 549th nucleotide, there were 4 ATG codons, two of which were in frame. The fourth ATG codon was in the best context to act as a start codon for encoding a protein which will be composed of 546 amino acids with a predicted molecular weight 59,310 Da. The comparison of the nucleic acid sequences of different morbillivirus F-protein genes revealed that the 5'-end sequence of the mRNA is specific to each virus. This sequence contains a long stretch of nucleotides rich in G/C content. When protein sequences were compared, it appears that, during evolution, substitutions in amino acid occurred in the F-protein of morbilliviruses such that the structure required for the fusion activity remains unchanged. These substitutions appear to have only occurred in the leader and the membrane anchor sequences, probably as an adaptation of the protein to the host cell. PMID- 7483820 TI - The cytoplasmic tail of the neuraminidase protein of influenza A virus does not play an important role in the packaging of this protein into viral envelopes. AB - We have rescued a transfectant influenza virus, NA/TAIL(-), whose neuraminidase (NA) protein lacks the predicted cytoplasmic tail. The virus was attenuated (one log10 reduction) both in tissue culture and in mouse lungs. Attenuation correlated with a 50% reduction of the level of NA in infected cells and levels of incorporation of the tail-less NA protein into viral particles paralleled that in infected cells. This result indicates that the signal for packaging of the NA protein into the viral envelope is not located in its cytoplasmic domain. PMID- 7483821 TI - Restriction endonuclease analysis and physical mapping of the genome of porcine adenovirus type 5. AB - The HNF61 and HNF70 isolates of porcine adenovirus type 5 (PAV-5) were cultivated in PK-15 cells, and viral DNA was extracted from the infected cells by a modified Hirt procedure. The DNAs were digested by each of 9 restriction endonucleases, and fragments representing the entire genomes were cloned. Based on the sizes of the restriction enzyme fragments, the genome of each isolate was estimated to be 33.2 kb. Physical maps for the 9 restriction endonucleases were constructed. The physical maps of the two isolates were identical for 5 of the restriction endonucleases, but 4 enzymes revealed differences in restriction sites occurring mainly between map units 78 and 83, which may include the E3 region of the genome. There were no similarities between the physical maps of PAV-5 and those described for the other 4 serotypes of PAV. PMID- 7483823 TI - Nucleoprotein phosphorylated on both serine and threonine is preferentially assembled into the nucleocapsids of measles virus. AB - The nucleoprotein (N) in the nucleocapsids of measles virus (MV) has different conformation and antigenicity than the free N-protein in MV-infected cells. These two forms of N-protein have identical methionine-containing tryptic peptides. The free N-protein contains 4 phosphorylated tryptic peptides. However, the nucleocapsid-associated N-protein has an additional phosphorylated peptide not found in the free N-protein. The free N-protein is phosphorylated only on serine residues, whereas the nucleocapsid-associated N-protein is phosphorylated on both serine and threonine residues. The MV N-protein expressed from a cloned gene in primate cells is also phosphorylated on both serine and threonine residues. These results suggest that cellular kinases phosphorylate the MV N-protein, and N protein with phosphorylated serine and threonine is preferentially assembled into the viral nucleocapsids. PMID- 7483822 TI - Identification of the major capsid protein gene of human herpesvirus 7. AB - Two human herpesvirus 6 (HHV-6) open reading frames were identified with significant amino acid similarity to UL86 (major capsid protein, MCP) and UL85 of human cytomegalovirus (HCMV) in human herpesvirus 7 (HHV-7) genes. The predicted lengths of the complete HHV-7 MCP and the HCMV-UL85 open reading frames were 1344 and 293 amino acids with estimated molecular weights of approximately 153 and 33 kDa, respectively. Computer analysis showed that the amino acid of HHV-7 MCP was 61% identical to the MCP of HHV-6 variants A and B, 28% to HCMV. These results suggest that HHV-7 is more closely related to HHV-6 than to HCMV. PMID- 7483824 TI - Characterization of 5'-upstream sequence of the latent membrane protein 1 (LMP-1) gene of an Epstein-Barr virus identified in nasopharyngeal carcinoma tissues. AB - Sequence variations of the 5'-upstream region of latent membrane protein 1 (LMP 1) in two Epstein-Barr virus (EBV) strains have been reported before (Chen et al., 1992). To investigate the effect of these variations on gene expression, we constructed a series of deletion plasmids encompassing positions -950 to +20 of the LMP-1 promoter region and tested for the ability to drive chloramphenicol acetyltransferase (CAT) gene expression in C33A cells. Results showed that the promoter activities of constructs from NPC strain were 3-fold lower than the corresponding constructs from the B95-8 strain. In addition, the region between 54 and +20 contained the basic, constitutive promoter activity for both strains. Sequence analysis of this region indicated that an activating transcription factor (ATF) binding site, TGACGTAG, which is present in B95-8 strain was changed to TCTCGTAG in NPC strain. A chimeric plasmid study suggested that these sequence variations in the ATF binding site may contribute to the 3-fold increase of CAT activity observed for B95-8 strain. Furthermore, the activity of the promoter constructs was not activated by EBV-encoded nuclear antigen 2 (EBNA-2) in C33A cells. However, the promoter activities were upregulated in B-lymphocyte cells such as CG3 and CA46 cells. The biological significance of this difference in promoter activity of LMP-1 gene between two strains and the involvement of the cellular factors were discussed. PMID- 7483826 TI - Properties of the active plum pox potyvirus RNA polymerase complex in defined glycerol gradient fractions. AB - As a first step in the study of the replication of plum pox virus (PPV) RNA, an in vitro virus-specific RNA polymerase activity was characterized in a crude membrane extract (Martin and Garcia, 1991). In this study, we report the fractionation of the crude membrane extract by centrifugation in glycerol gradients. The sedimentation properties after different treatments of the crude extract and its insensitivity to micrococcal nuclease treatment suggest that the RNA polymerase activity was localized in a defined and enclosed membranous structure. Subcellular membrane characterization of the different glycerol gradient fractions indicated that PPV-specific RNA synthesis occurred in fractions enriched in endoplasmic reticulum and tonoplast vesicles. PMID- 7483825 TI - gp13 (EHV-gC): a complement receptor induced by equine herpesviruses. AB - Equine herpesviruses type 1 (EHV-1) and type 4 (EHV-4) induce a complement receptor protein on the surface of infected cells capable of binding to the third component of complement (C3). The protein mediating the binding to the C3 component of complement was identified as glycoprotein 13 (gp13, EHV-gC), as expression of the cloned viral gene under the control of a CMV promoter induced C3 binding activity at the transfected cell surface. Comparable to glycoprotein C (gC) from herpes simplex virus type 1 (HSV-1-gC), glycoprotein III from pseudorabiesvirus (gIII, PRV-gC) and bovine herpesvirus-1 (gIII, BHV-1-gC), gp13 derived from EHV-infected cell lysates bound to C3 fixed to solid phase, showing preferential binding to the appropriate host complement component. Similar to wild-type isolates, a highly attenuated vaccine EHV-1 strain also displayed complement receptor activity despite apparent differences of the gp13 gene in restriction enzyme digest pattern and reactivity with monoclonal antibodies. In addition, other structural proteins were altered in the vaccine strain as compared to wild-type strains, which might contribute to its attenuated phenotype. In contrast to the situation observed with HSV-1-gC, the interaction of gp13 (EHV-gC) with horse complement was not inhibited by polyanionic substances like heparin or dextran sulfate. These results suggest structural differences in the particular binding mechanism of the respective viral envelope proteins. PMID- 7483827 TI - Alterations in physical state and expression of human papillomavirus type 18 DNA following crisis and establishment of immortalized ectocervical cells. AB - Integration of episomal human papillomavirus (HPV) DNA in infected cervical lesions during malignant progression is frequently observed, but the importance of integration is poorly understood. We have studied immortalization by HPV-18 of human cervical cells as an in vitro model system. Here, the status and expression of HPV-18 DNA in precrisis ectocervical keratinocytes was compared with that in the same cells after crisis and establishment of immortalization. Southern blots revealed, and two-dimensional gel analysis confirmed, that the precrisis culture contained more than 100 copies/cell of episomal HPV-18 DNA and no detectable integrated viral DNA. In contrast, the postcrisis cells contained a low copy number of only integrated viral genome. The Northern blot patterns of E6-E7 and E2/E4 RNA expression were also different. Analysis of RNA by RT-PCR indicated that neither culture expressed the unspliced HPV-18 E6 oncogene present in tumor cell lines and that the precrisis, but not postcrisis, culture expressed the full length E2 repressor. The two cultures displayed a similar keratinocyte morphology in vitro and a similar low grade dysplasia in vivo and both were non-tumorigenic. These results suggest that, although insufficient for complete malignant conversion, viral DNA integration during crisis is associated with the establishment of an immortalized phenotype in which HPV-18 DNA is integrated and HPV-18 RNA expression is altered. PMID- 7483828 TI - Electroporation of influenza virus ribonucleoprotein complexes for rescue of the nucleoprotein and matrix genes. AB - Reverse genetics has been successfully used for the generation of recombinant influenza virus with altered biological properties. The standard method is based on DEAE-dextran transfection of in vitro reconstituted influenza virus ribonucleoprotein complex (RNP) into helper virus infected cells with subsequent selection of the recombinant viruses. Here we report the utilization of electroporation for reverse genetics of influenza virus as an improvement over the standard method. In a neuraminidase (NA) gene rescue system, we were able to demonstrate that electroporation of in vitro reconstituted NA RNP of influenza A/WSN/33 (H1N1) virus into WSN/HK virus infected cells allows the rescue of the transfectant WSN virus. The titer of transfectant virus obtained using electroporation is comparable to that generated using the DEAE-dextran transfection method. More significantly, the ratio of transfectant virus to helper virus is as much as 20-fold greater than that achieved using the DEAE dextran system. We have also used electroporation to generate recombinant influenza virus carrying cDNA-derived matrix (M) gene or nucleoprotein (NP) gene of the WSN virus by using the temperature-sensitive (ts) mutants ts51 and ts56 as helper viruses. In the case of electroporation of M gene RNP, 88% of the viruses isolated after selection at 39 degrees C were transfectants. In contrast, the majority of viruses obtained using the DEAE-dextran transfection method were revertants of the helper virus. The NP-gene transfectant was only generated by the electroporation method. Our results suggest that electroporation of influenza virus RNP may be a useful method for generation of recombinant influenza viruses, especially in a system in which a ts mutant is used as helper virus. PMID- 7483829 TI - The vaccinia virus J5L open reading frame encodes a polypeptide expressed late during infection and required for viral multiplication. AB - A number of open reading frames (ORFs) are found in the vaccinia virus (VV) genome whose activities in the viral life cycle have not yet been determined. This report examines one such ORF, designated J5L, which was demonstrated to be essential for viral multiplication. Stable inactivation of the J5L ORF by insertion of a lacZ ORF was impossible unless another copy of the J5L ORF was present in the VV genome. Fusion genes between the J5L ORF and either the lacZ gene or the VV K1L gene were employed to study its temporal expression as well as its protein product. These experiments showed that J5L is transcribed late in infection and gives rise to a protein product which migrates by SDS-PAGE with the expected molecular weight (16 kDa). Numerous unsuccessful attempts to establish a stable cell line expressing J5L suggest that the J5L gene product could be cytotoxic. PMID- 7483832 TI - [Why does the DNA code contain 4 letters?]. AB - The answer to this question is not yet known. There are two ways to express information, i.e., to reduce the number of letters in alphabet (n), which simplifies the decoding machine, but leads to longer informational sequences, or to increase n, which shortens sequences, but complicates the informational machine. The compromise between these two possibilities would be to obtain the minimum of one of summary informational component's parameters. The summary component is the sum of corresponding decoding machine's and the program's parameters. In this work it was demonstrated that DNA four-letter code is optimal, for it allows the minimal volume of summary cell informational contents. But it is so only for the most simple DNA. Our calculations may indirectly show that such DNA (and not more complicated) was the object of "projecting" at one of the biological evolution's early stages. PMID- 7483831 TI - Cloning and sequence analysis of RNA-2 of a mechanically transmitted UK isolate of barley mild mosaic bymovirus (BaMMV). AB - A mutant of the 'Streatley' isolate of barley mild mosaic bymovirus was selected from the original field isolate by repeated mechanical inoculation. Unlike the wild-type barley mild mosaic virus, which is transmitted by the soilborne fungus Polymyxa graminis, the mutant could not be transmitted by this vector. RNA-2 of the mutant virus was shorter than that of the wild-type virus suggesting that a deletion of part of the genome segment had occurred. The nucleotide sequence of the mutant RNA-2 was determined and revealed a high degree of homology with the RNA-2 of a German BaMMV field isolate. The deletion comprises 1092 nucleotides and is located in the 3'-terminal part of the coding region. The 34-kDa truncated form of the C-terminal protein is less than half the size of the corresponding protein of full-length BaMMV RNA-2. Taken together, the sequence data and results of biological experiments suggest an essential role of the C-terminal protein for fungus transmission of BaMMV. PMID- 7483830 TI - Adenovirus protease expressed in insect cells cleaves adenovirus proteins, ovalbumin and baculovirus protease in the absence of activating peptide. AB - The adenovirus type 2 protease (EP) was expressed by infecting insect cells with a recombinant baculovirus. Immunoblot and activity analysis showed EP to be present in both the nucleus and cytoplasm. While the insect cell expressed EP was more soluble than the Escherichia coli expressed EP, its activity was one quarter of the latter, suggesting that eukaryotic postsynthetic modifications are not essential for enzyme activity. EP inactivated a cytoplasmic cathepsin-like baculovirus-encoded cysteine protease which carries a single EP cleavage site and which was capable of digesting most adenovirus structural proteins in vitro. In addition to cleavage of the baculovirus protease, the adenovirus EP was also able to cleave ovalbumin and canine adenovirus protein pre-VII, in the absence of activating peptide. EP activation therefore may occur by means of factors other than the specific activating peptide. PMID- 7483833 TI - [Biologically active substances in the tissues of hibernating animals]. AB - It was supposed that the transitional period from euthermia to hibernation rather than the period of winter hibernation is optimal for the secretion of biologically active substances in rodent tissues. This hypothesis was tested in experiment. The maximal O2 consumption suppression was noted in mice after I. P. Injection of blood plasma from suslik Citellus undulatus at the beginning of its entering into hibernation. Low-molecular peptide fraction appearance in C. undulatus blood before hibernation was revealed by electrophoresis in gel and autoradiography. But its traces disappear by the end of bout. The endogenic inhibitory factor's maximal influence in early bout was shown by experiments with TRH and neoklotorphin (NKT). I.P. injections of these substances to C. undulatus were ineffective in the beginning of entering into hibernation, but clearly caused awakening after hibernation, has started. KT, being a fragment of NKT, inhibited suslik's heart rate when the awakening was provoked in the middle, but not in the end of bout. Possible transformation of biologically active substances in winter-hibernating rodents tissues is discussed. The KT "inhibitor" and NKT "stimulator" are supposed to pertain to the special type of regulatory peptides, that manage the hibernation cycle. One of the ways of inactivation of endogenic "hibernation trigger" is its removal from the organism with urine. The urine, taken from suslik immediately after hibernation proved to produce the greatest hypothermic effect on mice, I.P. injected with it. PMID- 7483834 TI - Two-dimensional protein map of Saccharomyces cerevisiae: construction of a gene protein index. AB - This publication marks the beginning of the construction of a gene-protein index that relates proteins which are resolved on the two-dimensional protein map of Saccharomyces cerevisiae with their corresponding genes. We report the identification of 36 novel polypeptide spots on the yeast protein map. They correspond to the products of 26 genes. Together with the polypeptide spots previously identified, this raises to 41 the number of genes whose products have been identified on the protein map. The proteins identified here are concerned with four major areas of yeast cellular physiology: carbon metabolism, heat shock, amino acid biosynthesis and purine biosynthesis. Given the molecular weight and isoelectric point of the identified proteins, and the codon-usage bias of the corresponding genes, it can be estimated that 25 to 35% of all the soluble yeast proteins are detectable under the labelling and running gel conditions used in this study. PMID- 7483835 TI - Kluyveromyces lactis killer plasmid pGKL2: molecular analysis of an essential gene, ORF5. AB - The ORF5 of Kluyveromyces lactis killer plasmid pGKL2 (k2) is capable of encoding a small neutral protein of 18 kDa of as yet unassigned function. Although this ORF is located between two larger ORFs, 4 and 6, which it overlaps, RNA analysis showed that it is transcribed monocistronically. One-step gene disruption of ORF5, via in vivo homologous recombination between native plasmid k2 and a transfer vector employing the Saccharomyces cerevisiae LEU2 gene fused to the k2 UCS5 element, yielded Leu+ transformants at high frequencies. The transformants were found to carry a new recombinant form of k2 with ORF5 replaced by the LEU2 marker, termed rk2, in addition to the wild-type plasmids k1 and k2. Northern analysis detected a plasmid-dependent LEU2 transcript distinct in size and regulation from its nuclear counterpart. Recombinant plasmid, rk2, was unable to displace native k2 during Leu+ selective growth; however rk2 was displaced by k2 during non-selective growth. Thus, ORF5 appears to be an essential gene for plasmid integrity and/or maintenance. The ORF5 product was detected by over expression of an epitope-tagged allele in the baculovirus system. Western analysis using a monoclonal antibody specific for the epitope tag identified a protein band with apparent molecular weight of 20 kDa, corresponding in size to the predicted product. PMID- 7483836 TI - In vivo cloning by homologous recombination in yeast using a two-plasmid-based system. AB - In order to reduce the number of classical DNA manipulation and ligation steps in the generation of yeast expression plasmids, a series of vectors is described which facilitate the assembly of such plasmids by the more efficient 'recombination in vivo' technique. Two sets of vectors were developed. The first set, called 'expression vectors', contains an expression cassette with a yeast promoter and the PGK terminator separated by a polylinker, and an Escherichia coli replicon. Subcloning in these vectors of a DNA fragment generates a 'transfer vector' which is compatible with the second set of E. coli-yeast shuttle vectors. This set of 'recombination vectors' contains a cassette for a functional copy of a gene complementing a host strain auxotrophy or a bacterial gene conferring an antibiotic resistance to the plasmid-bearing host. Plasmid copy numbers can be modulated through the use of URA3 or URA3-d as the selective marker together with an ARS/CEN and the 2 microns replicon. Integration of the cloned DNAs into the yeast linearized replicative vectors occurs by recombination between homologous flanking sequences during transformation in yeast or E. coli. All the vectors contain the origin of replication of phage f1 and allow the generation of single-stranded DNA in E. coli for sequencing or site-directed mutagenesis. PMID- 7483837 TI - The maintenance of self-replicating plasmids in Saccharomyces cerevisiae: mathematical modelling, computer simulations and experimental tests. AB - A distributive model has been constructed to describe the maintenance of the native 2 microns and 2 micron-based plasmids in the yeast Saccharomyces cerevisiae. This model includes elements which represent the influence of selection, segregation, replication and amplification on plasmid stability. A computer program has been written in TURBO PASCAL to implement the model and a number of simulation experiments have been carried out. These simulations permitted the choice of a form of the model which is compatible with the available experimental evidence. The form chosen involves an amplification system in which the RAF gene product binds to the Rep1/Rep2 dimer to prevent the latter acting to repress the activity of the FLP gene. At the same time an upper limit (or 'ceiling') was imposed on the number of plasmid molecules able to replicate. Maternal bias was accommodated by 'tagging' a small proportion of molecules for inheritance by the mother nucleus and these tags being removed (or 'cleared') by the Rep1/Rep2 dimers. This final form of the model makes specific predictions about the stability of 2 microns and YEp plasmids in yeast populations and about the distribution of plasmid copy number between cells in such populations. The predictions on stability have been subjected to experimental test and results provide good support for the model. PMID- 7483838 TI - High-resolution cosmid mapping of the left arm of Saccharomyces cerevisiae chromosome XII; a first step towards an ordered sequencing approach. AB - For the sequencing of the left arm of chromosome XII of Saccharomyces cerevisiae, we fine-mapped the entire 450 kb fragment between the ribosomal DNA (rDNA) and the left telomere. Total yeast DNA in agarose blocks was digested with I-PpoI, which exclusively cuts once in each repeat unit of the rDNA. The resulting fragment was isolated from pulsed-field gels, together with the equally sized chromosome IX. A cosmid library of some 30-fold chromosome coverage was generated from this material, with the cloning efficiency being around 20,000 clones per microgram genomic DNA. The chromosome XII and IX specific clones were identified by complementary hybridizations with the respective chromosomes. For the left arm of chromosome XII, a contiguous cosmid array (contig) with an average map resolution better than 9 kb was generated by clone hybridization procedures. The ordered library serves as a tool for the physical mapping of genetic markers. Also, a minimal set of 15 clones was selected that covers the entire fragment. This subset forms the basis for the generation of a template map of much higher resolution for a directed sequencing of the left arm of chromosome XII. PMID- 7483839 TI - Sequence of a 9.8 kb segment of yeast chromosome II including the three genes of the MAL3 locus and three unidentified open reading frames. AB - We report the DNA sequence of a segment located on the right arm of chromosome II from Saccharomyces cerevisiae S288C near the subtelomeric sequences. The sequence was determined using a random cloning strategy followed by an oligonucleotide directed sequencing. The segment contains four non-overlapping open reading frames (ORFs) YBR297w, YBR298c, YBR299w and YBR301c, and two overlapping ones (YBR300c and YBR300w). Three of them--YBR297w, YBR298c and YBR299w--are the MAL3R (transcriptional regulatory protein), MAL3T (maltose permease) and MAL3S (maltase) genes of the MAL3 locus previously localized. The three other ORFs are unidentified. Another MAL locus (MALl) has been localized on chromosome VII. The Mal- phenotype of strain S288c cannot be explained by telomeric silencing. PMID- 7483840 TI - Analysis of a 32.8 kb segment of yeast chromosome IV reveals 21 open reading frames, including TPS2, PPH3, RAD55, SED1, PDC2, AFR1, SSS1, SLU7 and a tRNA for arginine. AB - We report the nucleotide sequence of a 32.8 kb DNA segment from the right arm of Saccharomyces cerevisiae chromosome IV. The sequence contains 20 open reading frames (ORFs) longer than 300 bp as well as the 240 bp gene coding for the essential SSS1 secretory protein. Nine ORFs previously totally or partially sequenced (TPS2, PPH3, RAD55, SED1, PDC2, AFR1, SSS1, SLU7 and D4478) are presented, as well as the transmembrane protein D4405, the leucine zipper containing D4495 and a new tRNA for arginine. D4456 and D4461 are separated by a single in-frame stop codon only. The other five ORFs show no particular features or significant homology. PMID- 7483841 TI - Sequence analysis of a 33.1 kb fragment from the left arm of Saccharomyces cerevisiae chromosome X, including putative proteins with leucine zippers, a fungal Zn(II)2-Cys6 binuclear cluster domain and a putative alpha 2-SCB-alpha 2 binding site. AB - In the framework of the European BIOTECH project for sequencing the Saccharomyces cerevisiae genome, we have determined the nucleotide sequence of the left part of the cosmid clone 232 and the cosmid clone 233 provided by F. Galibert (Rennes Cedex, France). We present here 33,099 base pairs of sequence derived from the left arm of chromosome X of strain S288C. This sequence reveals 17 open reading frames (ORFs) with more than 299 base pairs, including the published sequences for ARG3, LIGTR/LIG1, ORF2, ACT3 and SCP160. Two other ORFs showed similarity with S. cerevisiae genes: one with the CAN1 gene coding for an arginine permease, and one with genes encoding the family of transcriptional activators containing a fungal Zn(II)2-Cys6 binuclear cluster domain like that found in Ppr1p or Ga14p. Both putative proteins contain a leucine zipper motif, the Can1p homologue has 12 putative membrane-spanning domains and a putative alpha 2-SCB-alpha 2 binding site. In a diploid disruption mutant of ORF J0922 coding for the transcriptional activator homologue, no colonies appeared before 10 days after transformation and then grew slowly. In contrast, haploid disruption mutants showed a growth phenotype like wild-type cells. One ORF showed weak similarity to the rad4 gene product of Schizosaccharomyces pombe and is essential for yeast growth. Five ORFs showed similarity to putative genes on the right arm of chromosome XI of S. cerevisiae. Two of them have similarity to each other and belong to a family of extracellular proteins that groups mammalian SCP/Tpx-1, insects Ag3/Ag5, plants PR-1 and fungi Sc7/Sc14.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7483843 TI - Mapping of the ACC1/FAS3 gene to the right arm of chromosome XIV of Saccharomyces cerevisiae. AB - The ACC1/FAS3 gene has been mapped to the right arm of chromosome XIV by both genetic and physical methods. The gene is closely linked to RNA2 and is allelic to the ABP2 gene of chromosome XIV. PMID- 7483842 TI - Localization of the FAR3 gene: genetic mapping and molecular cloning using a chromosome walk-'n'-roll strategy. AB - FAR3 is a newly-discovered yeast gene required specifically for pheromone mediated cell cycle arrest. I have used strains harboring the far3-1 mutation to map the gene to the right arm of chromosome XIII, establishing the gene order CEN13-LYS7-MCM1-FAR3. I cloned the FAR3 gene based on its genetic map position using a strategy that combined chromosome walking and a related technique termed 'chromosome rolling'. In addition to the genetic and physical localization of FAR3, I present data that suggest corrections to the tentative map positions of VAN1 and ARG80. PMID- 7483845 TI - The Saccharomyces cerevisiae FLO1 flocculation gene encodes for a cell surface protein. AB - The sequencing of a 6619 bp region encoding for a flocculation gene previously cloned from a strain defined as FLO5 (Bidard et al., 1994) has revealed that it was a FLO1 gene. The FLO1 gene product has been localized at the cell surface of the yeast cell by immunofluorescent microscopy. The Flo1 protein contains four regions with repeated sequences which account for about 70% of the amino acids of this protein. A functional analysis of the major repeated region has revealed that it plays an important role in determining the flocculation level. A gene disruption experiment has shown that FLO5 strain STX 347-1D contains at least two flocculation genes of the FLO1 type but that they are supposed to be inactive and do not contribute to its flocculation. However, enzyme-linked immunosorbent assays performed on intact cells have revealed that a protein expressed at the cell surface of the FLO5 strain STX 347-1D is antigenically related to Flo1p. A deletion analysis of the 5' region of the FLO1 gene has shown that the expression is submitted to controls which depend on the genetic background of the strain. PMID- 7483846 TI - The red ade mutants of Kluyveromyces lactis and their classification by complementation with cloned ADE1 or ADE2 genes from Saccharomyces cerevisiae. AB - Seventy-six red adenine mutants of Kluyveromyces lactis were isolated. By complementation they could be assigned to two groups with 31 and 45 mutants. Transformation of several strains from each group with plasmids containing the Saccharomyces cerevisiae ADE1 or ADE2 gene showed that the largest group was ade2 and the other group was ade1. Several previously isolated 'ade1' mutants were classified to either group and given new gene and allele numbers. ADE1 was localized at chromosome III, closely linked to the mating type gene, making it a convenient marker for mating type. ADE2 was localized at chromosome V. PMID- 7483844 TI - Biochemical similarity of Schizosaccharomyces pombe ras1 protein with RAS2 protein of Saccharomyces cervisiae. AB - Schizosaccharomyces pombe contains single ras oncogene homologue, ras1, that functions in the signal transduction pathway conducting the cell's mating processes. To understand the biochemical basis of yeast ras proteins, we have purified the ras1 protein and compared the major biochemical constants with those of RAS2 protein from Saccharomyces cerevisiae and mammalian ras proteins. The purified ras1 protein showed a remarkably high Kd value for GDP binding (178 nM) and for binding with ATP. In contrast, the Kd value for GTP binding and the rate of GTPase activity were 64 nM and 77 x 10(-6) s-1 at 37 degrees C, respectively; both were higher than normal p21ras protein, but at the same level as the RAS2 protein. We directly measured rate of GTP binding and GDP binding which were 3.9 x 10(-3) s-1 and 1.8 x 10(-3) s-1 at 30 degrees C, respectively. On the other hand, exchange rates between bound and free nucleotides remained almost constant throughout the tested combination of GTP and GDP, and were several-fold lower than the binding rate. These results suggest that the release of the guanine nucleotide is the rate-limiting step in the ras-GTP/GDP cycle. As a whole, the biochemical properties of the ras1 protein are close to those of the RAS2 protein, although these two proteins function differently in the signal transduction pathway in the cells. PMID- 7483849 TI - Expression and secretion of antifreeze peptides in the yeast Saccharomyces cerevisiae. AB - The antifreeze peptide AFP6 from the polar fish Pseudopleuronectus americanus has been expressed in and secreted by the yeast Saccharomyces cerevisiae as a biologically active molecule. The gene for the 37 amino acid long peptide has been chemically synthesized using yeast preferred codons. Subsequently, the gene has been cloned into an episomal expression vector as well as in a multicopy integration vector, which is mitotically more stable. The expression is under the control of the inducible GAL7 promoter. The enzyme alpha-galactosidase has been investigated as a carrier protein to facilitate expression and secretion of AFP. In order to reach increased expression levels, tandem repeats of the AFP gene (up to eight copies) have been cloned. In most cases the genes are efficiently expressed and the products secreted. The expression level amounts to approximately 100 mg/l in the culture medium. In a number of genetic constructs the genes are directly linked and expressed as AFP multimers. In other constructs linker regions have been inserted between the AFP gene copies, that allow the peptide to be processed by specific proteinases, either from the endogenous yeast proteolytic system or from a non-yeast source. The latter requires a separate processing step after yeast cultivation to obtain mature AFP. In all these cases proteolytic processing is incomplete, generating a heterogeneous mixture of mature AFP, carrier and chimeric protein, and/or a mixture of AFP-oligomers. The antifreeze activity has been demonstrated for such mixtures as well as for AFP multimers. PMID- 7483847 TI - Characterization of Na+/H(+)-antiporter gene closely related to the salt tolerance of yeast Zygosaccharomyces rouxii. AB - In order to clarify the relationship between salt-tolerance of Zygosaccharomyces rouxii and the function of Na+/H(+)-antiporter, a gene was isolated from Z. rouxii which exhibited homology to the Na+/H(+)-antiporter gene (sod2) from Schizosaccharomyces pombe. This newly isolated gene (Z-SOD2) encoded a product of 791 amino acids, which was larger than the product encoded by its Sz. pombe homologue. The predicted amino-acid sequence of Z-Sod2p was highly homologous to that of the Sz. pombe protein, but included an extra-hydrophilic stretch in the C terminal region. The expression of Z-SOD2 was constitutive and independent of NaCl-shock. Z-SOD2-disruptants of Z. rouxii did not grow in media supplemented with 3 M-NaCl, but grew well in the presence of 50% sorbitol, indicating that the function of Z-SOD2 was closely related to the salt-tolerance of Z. rouxii. Several genes are also compared and discussed in relation to the salt-tolerance of Z. rouxii. PMID- 7483848 TI - A short-chain dehydrogenase gene from Pichia stipitis having D-arabinitol dehydrogenase activity. AB - An NAD(+)-dependent D-arabinitol dehydrogenase (polyol dehydrogenase) gene was isolated from Pichia stipitis CBS 6054 and cloned in Saccharomyces cerevisiae. The gene was isolated by screening of a lambda-cDNA library with a zymogram technique. D-Arabinitol, xylitol, D-glucitol and galactitol are substrates for the recombinant protein. With D-arabinitol as substrate the reaction product is D ribulose. The molecular weight of the native tetramer enzyme is 110,000 Da and the monomer is 30,000 Da. The amino acid sequence is homologous to the short chain dehydrogenase family. It is 85.5% identical to a D-arabinitol dehydrogenase from Candida albicans. The gene in P. stipitis was induced by D-arabinitol and P. stipitis was able to grow on D-arabinitol. The physiological role of D-arabinitol metabolism is discussed. PMID- 7483850 TI - Sequence and functional analysis of a 7.2 kb DNA fragment containing four open reading frames located between RPB5 and CDC28 on the right arm of chromosome II. AB - In a coordinated approach, several laboratories sequenced Saccharomyces cerevisiae chromosome II during the European BRIDGE project. Here we report on the sequence and functional analysis of a 7217 bp fragment located on the right arm of chromosome II between RPB5 and CDC28. The fragment contains four open reading frames probably encoding proteins of 79.2 kDa (corresponding gene YBR156c), 12.1 kDa (YBR157c), 62.7 kDa (YBR158w) and 38.7 kDa (YBR159w). All four open reading frames encode new proteins, as concluded from data base searches. The respective genes were destroyed by gene replacement in one allele of diploid cells. After sporulation and tetrad analysis, the resulting mutant haploid strains were investigated. No phenotype with respect to spore germination, viability, carbohydrate utilization, and growth was found for YBR157c, encoding the smallest open reading frame investigated. Gene replacement within the YBR156c gene encoding a highly basic and possibly nuclear located protein was lethal. Ybr158 revealed similarities to the Grrl (Cat80) protein with respect to the leucine-rich region. Cells harboring a mutation in the YBR158w gene showed strongly reduced growth as compared to the wild-type cells. The protein predicted from YBR159w shared 33% identical amino acid residues with the human estradiol 17 beta-hydroxysterol dehydrogenase 3. Haploid ybr159c mutants were only able to grow at reduced temperatures, but even under these conditions the mutants grew slower than wild-type strains. PMID- 7483853 TI - Current awareness on yeast. PMID- 7483854 TI - [International Journal of Food Research and Technology]. PMID- 7483852 TI - A new essential gene located on Saccharomyces cerevisiae chromosome IX. AB - A new 1150 amino acids long open reading frame (ORF), coding for an essential protein of unknown function was found in Saccharomyces cerevisiae by sequencing 3754 bp of geonomic DNA. The clone was isolated in a search for a fatty acid binding protein (FABP) and was localized on chromosome IX. The ORF bears no homology to FABP, but it shows weak similarity to Plasmodium vivax reticulocyte binding protein 1 and to aggregation-specific adenylate cyclase from Dictyostelium discoideum. The new gene is constitutively transcribed regardless of the carbon source used. PMID- 7483851 TI - A 37.5 kb region of yeast chromosome X includes the SME1, MEF2, GSH1 and CSD3 genes, a TCP-1-related gene, an open reading frame similar to the DAL80 gene, and a tRNA(Arg). AB - The complete DNA sequence of cosmid clone p59 comprising 37,549 bp derived from chromosome X was determined from an ordered set of subclones. The sequence contains 14 open reading frames (ORFs) containing at least 100 consecutive sense codons. Four of the ORFs represent already known and sequenced yeast genes: B645 is identical to the SME1 gene encoding a protein kinase, required for induction of meiosis in yeast, D819 represents the MEF2 gene probably encoding a second mitochondrial elongation factor-like protein, D678 is identical to the yeast GSH1 gene encoding gamma-glutamylcysteine synthetase and B746 is identical to the CSD3 gene, which plays an as yet unidentified role in chitin biosynthesis and/or its regulation. The deduced amino acid sequence of A550 is 63% identical to the Cc eta subunit of a murine TCP-1-containing chaperonin and more than 35% identical to thermophilic factor 55 from Sulfolobus shibatae, as well as to a number of proteins belonging to the chaperonin TCP-1 family. Open reading frame F551 exhibits homology to two regions of the DAL80 gene located on yeast chromosome XI encoding a pleiotropic negative regulatory protein. In addition, extensive homology was detected in three regions including parts of ORFs A560, B746/CSD3 and the incomplete ORF C852 to three consecutive ORFs of unknown function in the middle of the right arm of chromosome XI. Finally, the sequence contained a tRNA(Arg3) (AGC) gene. PMID- 7483857 TI - [Rapid determination of the value-determining components in sausage products by near-infrared transmission spectroscopy]. AB - A rapid method for the simultaneous determination of water, fat, crude protein, connective-tissue-protein and connective-tissue-protein free meat protein in sausage products with the help of near-infrared transmission spectroscopy (NIT) is presented. The calibrations for the each constituent were achieved with the chemometric method "partial least squares" (PLS) and validated with the added samples. Correlation coefficients between the reference methods and NIT were > or = 0.95 for water, fat, crude protein and connective-tissue-protein frei meat protein, and > or = 0.86 for connective-tissue-protein. The precision of NIT was comparable with the reference methods. PMID- 7483855 TI - Selenium content of breast milk. AB - Selenium levels in human milk in the winter period ranged from 5.3 micrograms/l to 23.8 micrograms/l, the mean value being 11.0 micrograms/l. The nursing women were divided into several groups according to the results of a questionnaire, i.e. according to their social status (refugees or otherwise), number of deliveries, post partum days, the weight they had gained during pregnancy, their age and smoking habits. The mean levels of selenium for each group are presented. Selenium was determined by hydride generation atomic absorption spectrometry. PMID- 7483856 TI - Measurement of the daily dietary calcium and magnesium intake in Belgium, using duplicate portion sampling. AB - The daily dietary calcium and magnesium intakes in Belgium were evaluated by sampling duplicate portions of food, destruction in a microwave oven and their calcium and magnesium contents determined by atomic absorption spectroscopy. The mean intake value for calcium (618 +/- 125 mg/day) was found to be less than those values measured in most other countries and was below the recommended daily allowance (RDA). The mean intake value for magnesium (271 +/- 44 mg/day) was similar to levels found in most other countries, but was below the RDA value for healthy adult men (350 mg/day) whilst being adequate for healthy women (280 mg/day). PMID- 7483858 TI - Physico-chemical properties and rheological behaviour of Patella caerula paramyosin. AB - Paramyosin is a muscle protein which is characteristic of all invertebrates but which is not present in vertebrate muscles. Given the functional importance of paramyosin, the purpose of this paper was to study the physico-chemical properties, including the amino acid composition and rheological behaviour, of purified paramyosin and to investigate its mode of interaction with myosin. Paramyosin was purified from the limpet (Patella caerula) by an ethanol precipitation step. It was soluble at ionic strengths below 0.05 M NaCl and its maximum solubility at neutral pH occurred at approximately 0.4 M NaCl. At this high ionic strength, the pH dependence of solubility was such that paramyosin passed quickly into solution when pH exceeded pH 5, the transitional pH value. By using an immunological method, it was shown that interactions between paramyosin and myosin occurred, even in the presence of actin. The molecular assembly of both proteins was probably specified by hydrophobic interactions, as well as by interactions enhanced by divalent cations. The changes in the dynamic shear storage modulus (G') started between 40 degrees C and 50 degrees C, and reached a maximum at about 75 degrees C. PMID- 7483859 TI - Static headspace gas chromatography of acetaldehyde in aqueous foods and polythene terephthalate. AB - Polythene terephthalate (PET) is frequently used as a packaging material for mineral water and other non-alcoholic beverages. PET contains detectable amounts of acetaldehyde, which is able to migrate to its packed product. An automated headspace gas chromatographic method for the determination of acetaldehyde has been developed and was used for the quantification of acetaldehyde in aqueous food products and their PET packages. A cold trap, mounted in the GC oven, between the auto sampler and analytical column was introduced as a new application. The detection limit of acetaldehyde was found to be 3 ng/ml with a standard deviation of 3%. The contents of acetaldehyde found in carbonated mineral water and lemonade ranged between 11 ng/ml and 7447 ng/ml, while the contents of acetaldehyde in the PET packages ranged from 1.1 microgram/g to 3.8 micrograms/g. PMID- 7483860 TI - [Automated pre-column derivatization with o-phthaldialdehyde (OPA)> A new RP-HPLC method for the determination of biogenic amines in food]. AB - A simple, selective and highly sensitive HPLC method for the routine determination of the biogenic amines in food is presented. Sample preparation is based on a rapid amine extraction using 10% trichloroacetic acid and a cation exchange column for extract purification. For the RP-HPLC analysis OPA/2 mercaptoethanol is used for the pre-column derivatisation, followed by fluorescence detection (Ex 345 nm, Em 440 nm). The effects of several factors are discussed. A separation of 15 biogenic amines is achieved within 70 min. The recoveries for histamine, tyramine, putrescine, cadaverine, tryptamine and beta phenylethylamine are higher than 95%. The detection limits lie between 0.1-0.5 pMol/injection (20 microliters), depending on the amine and a good linearity is achieved in the range from 0.5-500 pMol (r > 0.99). The method has been applied for the determination of biogenic amines in swiss cheese, salami, milk, beer and wine, the repeatability is very good. PMID- 7483862 TI - HPLC analysis of tonka bean extracts. AB - Based on the results of GC-MS analysis, an HPLC procedure was developed for the determination of coumarin, dihydrocoumarin, melilotic acid (o-dihydrocoumaric acid), methyl melilotate, ethyl melilotate, 5-hydroxymethylfurfural and o coumaric acid which are characteristic compounds of tonka beans. The presence of melilotic acid and ethyl melilotate in tonka beans has not been reported previously. Following HPLC analysis of tonka bean extracts produced in the laboratory and one sample of commercially available tonka bean absolute, evidence was provided that solvents used for production are decisive for the kind and amount of the compounds detected. PMID- 7483861 TI - [Analysis of some neuroleptics and carazolol in pork kidneys]. AB - A method is described for the analytical determination of the neuroleptics azaperone (plus its metabolite azaperol), acetylpromazine, propionylpromazine, chlorpromazine and the beta-blocking agent carazolol in pork kidneys. These compounds may be used illegally to calm pigs during their transport to the abattoir. The kidney samples (plus atosil as an internal standard) are incubated with NaOH (90 degrees C, 60 min); the rather fluid samples are extracted with diethylether. The separation of interfering compounds in the extracts is achieved on a silica gel column. The remaining interfering compounds are removed with ether after acidification of the eluted material. After alkalization of the aqueous solution, the drugs are extracted with either and applied to a Symmetry RP18 column (mobile phase: acetate buffer solution pH 4.5/acetonitrile/tetrahydrofurane, 65/30/10 by volume by HPLC. Recovery in spiked samples of pork kidneys (recovery of 50 micrograms/kg, carazolol 5 micrograms/kg) was between 71.5% (for propionylpromazine) and 88% (for azaperol). The method was verified on samples of treated pigs. PMID- 7483863 TI - Effect of irradiation on the microbiological status and flavouring materials of selected spices. AB - Spices from Egyptian local markets were irradiated with different recommended doses (0, 5, 10, 20 and 30 kGy). The spices tested included dried leaves of marjoram (Majorana hortensis Moench), rhizomes of ginger (Zingiber officinale Roscoe) and powdered hot pepper (Capsicum annum L.). The study included the isolation and identification of micro-organisms in spices following their irradiation, as well as gas chromatographic (GLC) chemical analysis for the presence and structure of volatile oils, pungent and pigment materials. The results showed that hot pepper was contaminated more (9.2 x 10(5)/g) than marjoram (4.2 x 10(3)/g) and ginger (14.3 x 10(3)/g) with respect to total aerobic bacterial content. The total contents of moulds were 4.8 x 10(3)/g, 5.7 x 10(3)/g and 19 x 10(3)/g in the same spices, respectively, but the pathogenic moulds and bacterial strains differed according to the type of spice. Irradiation at 10, 20 and 30 kGy caused complete elimination of micro-organisms, whereas 5 kGy was less effective. With the GLC method chosen 18 and 50 compounds could be detected in the extracts of marjoram and ginger, respectively; gamma-terpinen and zingiberen being the major compounds in marjoram and ginger, respectively. A noticeable reduction was observed in the amount of terpenes present in irradiated marjoram; they were converted to monoterpenesalcohols. Ginger was more sensitive to irradiation, especially at high doses, but moderate changes were detected at low doses (5 and 10 kGy). A slight, but significant effect on the capsaicin (pungent compound) in hot-pepper was observed following irradiation, whereas no changes in total pigments resulted at any dose. These results prove that 10 kGy is a sufficiently high dose to eliminate the micro-organisms in spices, causing only slight changes in the flavouring materials. PMID- 7483865 TI - [Diagnosis and therapy of "malnutrition" risk factors]. PMID- 7483864 TI - [Determination of Escherichia coli Shiga-like toxins by means of the MTT bioassay]. AB - Tissue culture cells' metabolism and viability are measured by the mitochondrial reduction rate of a yellow tetrazolium salt (MTT) to blue formazan crystals in the MTT-bioassay. Thus the MTT-bioassay is a standardizable and reproducible bioassay for measuring cytotoxicity or cytostimulation. It is shown that the MTT bioassay is also very suitable for determining bacterial cytotoxins using Escherichia coli's Shiga-like toxins as example. 177 strains of E. coli, isolated from carcasses and organs of cattle, are classified biochemically and tested for cytotoxin production by means of the MTT-bioassay. One of these strains is recognized as producer of Shiga-like toxin 2. 4 Enterohemolysin-producing strains of E. coli are cultivated from a feces sample of a diarrhoeic nubian ibex and identified as Shiga-like toxin 1 producers by help of the MTT-bioassay. PMID- 7483866 TI - [Gastroduodenal stress lesions and hemorrhages--pathogenesis, diagnosis, prevention and therapy]. AB - Following a short summary of the historical development of the terminology, the nosological separation between peptic ulcer disease and secondary ulcers including the gastroduodenal stress erosions and ulcers - is stressed. The differing and in the last 20 years decreasing frequency of the different causes as well as the pathogenesis - mainly disturbances of the mucosal defense mechanisms (blood supply, mucus secretion and epithelial renewal) - are discussed, partly referring to results of own investigation. The diagnosis implies - depending on the state of the patient - an emergency esophagogastroduodenocopy after the treatment of shock. The different outcome of bleeding in common peptic ulcer and in stress lesions is analyzed in detail as well as the limiting risk factors. The decrease of the incidence and the improvement of prognosis under a modern interdisciplinary intensive care of acutely ill patients, especially due to a strict shock therapy are stressed. The interventional-local therapy of stress-induced gastric or duodenal bleeding is not practicable, debatable, if performed with drugs, and surgically not possible; however, this dilemma is as mentioned above due to the less frequent occurrence of such a situation considerably diminished. The major effort of prophylaxis of gastroduodenal stress lesions is clearly directed towards an influence of causes and risks; several drugs - mainly H2 blocking agents, sucralfate and antacids demonstrate a comparable efficacy. The potential risk of a nosocomial pneumonia is differently seen; undebatable trials demonstrating a clear superiority of either prophylactically applied drugs do not exist. PMID- 7483867 TI - [Preoperative status of serum proteins in lung surgery patients]. AB - The preoperative serum-protein-status of 91 patients was investigated with a laboratory-screening in order to find out the patients with a malnutrition and consequently with a high risk of postoperative complications. The diagnosis was the basis for the classification in groups. There were no significant differences between values of our patients and normal values of a control group in protein, albumin, prealbumin, transferrin, cholinesterase, alpha-1-antitrypsin and haemopexin. Nutrition parameters are of limited value of preoperative risk screening in operable lung cancer patients. PMID- 7483870 TI - [Laparoscopic ligamentum teres-plasty]. AB - Report on 40 patients suffering from a therapy resistant gastroesophageal reflux disease. Conventional ligamentum-teres-plastic was performed on 18 patients, while 22 patients were treated laparoscopically as minimal invasive procedure. Both techniques are described and the advantages of the laparoscopy versus the conventional laparotomy are explained. As a result the ligamentum-teres-plastic is an efficient treatment for gastroesophageal reflux disease and distinguishes itself because of its relative simplicity and a low complication rate. PMID- 7483869 TI - [Laparoscopic sigmoid resection]. AB - Laparoscopic colorectal surgery not only presupposes knowledge in conventional surgery of the large bowel but also requires extensive experience in laparoscopic techniques. Furthermore every individual of the operating team should have suitable experience to guarantee effective co-operation. The presented study reports on 41 laparoscopic resections of sigmoid colon and upper rectum. The operations were carried out on 31 patients with benign and 10 patients with malignant diseases. In 1 case we had to change to an open procedure. Postoperative complications were seen in 10 patients, one of whom died (secondary haemorrhage 2; anastomotic leakage 1, Douglas haematoma 1, inflammation of the drainage tract 6). Preoperative management, intraoperative procedures and postoperative results are described and discussed. PMID- 7483868 TI - [Postoperative infusion therapy: electrolyte solution in comparison with hypocaloric glucose and carbohydrate exchange-amino acid solutions]. AB - OBJECTIVES: Comparison of early postoperative hypocaloric nutrition with glucose or xylitol/sorbitol- amino acid solutions versus saline alone. Influence on substrate, especially protein metabolism. DESIGN: Prospective randomized study. SETTING: Intensive care unit of an university hospital. PATIENTS: 44 patients in three groups after major surgery with necessity of intensive care. RESULTS: Hypocaloric infusions were well tolerated immediately after operation and positively influenced functional proteins and N-balance. During saline infusion protein parameters deteriorated significantly already after 24 to 36 hours. Glucose was not inferior to the xylitol/sorbitol solution and both resulted in a stable metabolic state during four days. CONCLUSIONS: After major trauma or surgery hypocaloric nutrition should be started within 24 hours and can be given already directly after the operation period without metabolic deterioration. PMID- 7483871 TI - [Adrenal incidentaloma]. AB - Asymptomatic adrenal tumors are discovered more and more frequently with improving quality of diagnostic imaging. Treatment of these tumors, described as incidentalomas is controversial and depends on the one hand on the size of the tumor and on the other hand on morphological appearance in diagnostic imaging. In the literature a tumor size over 3 cm is in general regarded as an indication for operation, based on findings of Copeland et al. where the size of the tumor was correlated with malignancy. In the Department of Surgery in Mannheim 28 operations on adrenal incidentalomas have been performed between 1973-1993. In three patients a carcinoma was found. Four of the removed tumors each of them benign had a diameter smaller than three centimeters. An operation seems to be justified under certain conditions, even in small tumors, because potentially malignant tumors can undergo early and radical surgery. A general recommendation for treatment of the above mentioned tumors, depending only on size and morphology in diagnostic imaging, seems to be difficult. The decision for surgical therapy needs to be evaluated individually for every patient. PMID- 7483872 TI - [Surgical therapy of bone metastases]. AB - The surgical treatment of bone metastases is generally palliative. All operative procedures have to consider the mostly short life expectancy and therefore have to guarantee stability, early mobilization and short hospitalization. Surgery is absolutely indicated in patients with pathologic fractures of the long bones and innominate bone involving the hip joint, in patients with spinal lesions causing instability and progressive neural deficit and in patients with compression of peripheral nerves. Relative indications for surgery are given in cases where the risk of sustaining a pathologic fracture is high, in cases where spinal neurological deficit or root irritation can be expected, in patients where the tumor grows despite adequate irradiation or chemotherapy and finally in cases where open biopsy is useful to exclude primary malignant tumor or is necessary to define the further therapeutic management. As morbidity, hospitalization and complications increase in patients with pathologic fractures prophylactic stabilization is emphasized using well defined criteria. The standard surgical principles and techniques are presented and the results of 506 cases are reported. PMID- 7483873 TI - [Long-term results of cemented total endoprostheses of the hip joint]. AB - 211 Muller-Charnley resp. Muller-Straight-Stem prostheses have been implanted from 1974 to 1984 in our Department for Trauma-Surgery. They were evaluated 11.8 years after implantation by clinical and radiological examination. 24 (12.8%) prostheses have been replaced because of aseptic loosening. Patients have been further evaluated by means of a new score, which emphasizes the mobility of the patients after total hip replacement. Mobility decreases significantly in patients over 70 years. PMID- 7483875 TI - [What are the advantages of UTN (unreamed tibial nail) in open tibial fractures?]. AB - The AO-unreamed tibial nail (UTN) is an alternative to external fixation in the treatment of open tibial fractures. In order to compare the two implants we did a retrospective study. 70 unselected cases of open tibial fractures were treated by either method. 35 patients with primary UTN were compared to another 35 patients with primary external fixator. Demographic data, localisation and classification of fractures and soft tissue trauma showed no differences between the two groups. The main complications in UTN-group were hardware problems, in external fixator group infections. Following UTN-treatment duration of first hospital stay and whole fracture treatment were shortened with earlier full weight bearing and fracture healing. Our study shows advantages in the treatment of open tibial fractures using the UTN compared to the external fixation. PMID- 7483874 TI - [Hemiprosthesis in femoral neck fracture]. AB - From 1968 to 1993 835 hemiarthroplasties were performed at the Department of Trauma Surgery of the University of Erlangen. Until 1984 Moore (297) and Thompson (148) prostheses were implanted. After this period 390 patients were treated with a bipolar headneck replacement. Average age of patients at the time of surgery was 80.2 years from 1968 to 1980 and 83.1 from 1981 to 1993. All hemiarthroplasties were performed due to intracapsular fractures of the femoral neck. 5.9% of the Thompson and Moore prostheses were revised to total hip arthroplasty, the rate of revisions of bipolar prostheses was 4.9%. Main causes of revision for Thompson and Moore prostheses were acetabular erosion and protrusion (5.2%). Revision of bipolar prostheses became necessary because of recurrent luxation in 4.1%. The social status of the patients was examined 1 year after the operation. From 1968 to 1980 67% of the patients lived in their own home or with their families, compared to 39% from 1981 to 1993. The percentage of patients in retirement homes tripled from 9% to 29% for the same period. PMID- 7483876 TI - [Treatment of supracondylar femoral fracture proximal to a knee joint endoprosthesis by retrograde interlocking nailing]. AB - A case of a supracondylar femoral fracture proximal to a total knee prosthesis is reported. This difficult fracture was managed with retrograde intramedullary locked nailing. The time to union was 6 weeks. Cancellous bone grafting was not necessary. At final follow up the patient had a knee score of 90 and a functional score of 80 points. This was equal to the result after joint replacement prior to the injury. PMID- 7483877 TI - [Nonelastic deformations of polyethylene inlays in hemiprostheses]. AB - In most models of hip prosthesis, the inlay is formed by a layer of polyethylene. The attrition of polyethylene results from two components, wear and nonelastic deformation. Former studies analysed total attrition. The aim of this trial was to measure solely the non elastic deformation. Nonelastic deformation was investigated using two different types of bipolar hemiprostheses to exclude influences of different shaped metal caps. The bipolar hemiprostheses were tested with a hydraulic pulser. 1 x 10(6) cycles of a halfsinewave with a stress of 4600 N at a frequency of 8 Hz were used. The whole procedure was carried out in isotonic saline solution. Stress was applied along a single axis to prevent any wear. After the test, the inner surface of the polyethylene inlay was analysed with a touchpeg. As the inlays had different inner diameters, the analysis of the surface was defined along radials of 5 degrees, 25 degrees and 45 degrees. The nonelastic deformation in the direction of the main stress at 45 degrees was higher than at the lateral areas. The nonelastic deformation measured was between 0.2 and 0.6 mm/a, i.e. 5 to 30% of total attrition. There was a significant difference between the two types of inlays. The model with several pegs and edges for a complicated snap mechanism had a significant higher nonelastic deformation (alpha = 0.05) than the smooth shaped inlay. PMID- 7483879 TI - [Danazol for treatment of fibrocystic mastopathy]. AB - 59 patients with extensive mastopathia, aged between 23 and 58 years, had been examined over a period of approximately 36 months. They were treated with danazol (Winobanin, Sanofi-Winthrop, Munich) at doses between 100 and 800 mg/day because of an extensive and partly persisting mastopathia. The documented period of treatment ranged between 40 and 774 days. According to the chosen medium daily dose the patients were divided into three groups (< 400 mg/d, 400-600 mg/d, > 600 mg/d). These groups were then subdivided according to criteria of symptoms, palpation findings, mammography and sonography. Between the onset and the end of treatment there was a significant improvement of mastopathia up to a complete recovery in more than 60% of the women. There were no significant systematic differences between the various dosages of danazol. In this respect it was impossible to verify assumed relation between dosage and success of treatment according to anamnestic or demographic factors. The objective evaluation by mammography and ultrasound showed an improvement in 81.4% of the patients, demonstrated by a decreased density of parenchyma and an increased subcutaneous fatty tissue of the mammae. The subjective success of therapy of the patients concerned was judged to be good in 75% of the cases by the doctor and in more than 63% by the patient herself. In conclusion, this study confirms that danazol offers very good therapeutic results for the treatment of the frequently persisting benign breast disease. PMID- 7483878 TI - [Adhesion and penetration of human lymphocytes through allogeneic endothelial cells within the scope of organ rejection]. AB - INTRODUCTION: Cellular rejection mechanisms are characterized by the infiltration of sensitized lymphocytes through the donor endothelium into the transplanted organ. The regulation of cellular adhesion molecules by soluble mediators (cytokines) is thought to play a dominant role in this process. In the present study the kinetics of lymphocyte infiltration were examined using a new established in vitro model and compared to the kinetics of adhesion molecule expression. MATERIALS AND METHODS: Peripheral blood lymphocytes (PBL) of healthy volunteers were pipetted to allogeneic endothelial cell (EC) monolayers and binding rates evaluated after different incubation times. Adherent PBL could be distinguished from penetrated PBL by means of a combined phase contrast- / reflection interference contrast - microscope. RESULTS: 30-35% of all pipetted PBL adhered to unstimulated EC maximally. Out of these cells < 10% penetrated through the endothelium ( = maximum penetration). The cytokines alpha-, beta-, gamma-interferon (IFN) or IL-1 did not enhance maximum adhesion, but accelerated this process. However, a 2 hrs prestimulus of EC by these cytokines was necessary to induce acceleration. Maximum penetration was enhanced by alpha-, beta-, gamma IFN, but not by IL-1, irrespective whether PBL were added together with cytokines to unstimulated EC or to already prestimulated EC. Immunocytochemical and fluorometric analyses of adhesion molecule expression revealed a cytokine induced upregulation or de novo expression of the adhesion antigens ICAM-1, ELAM-1 and VCAM-1 on EC membranes. Interestingly, PBL adhered to EC before upregulation or de novo expression of adhesion molecules was detected. DISCUSSION: The results showed that PBL adhesion and penetration processes were regulated differentially by cytokines. The early phase of PBL attachment to EC seemed not to be influenced by adhesion antigens but by an activation of the lymphocyte cytoskeleton. PMID- 7483880 TI - [Reactive and areactive actinomycosis infection of the female genitals and differentiation of pseudoactinomycosis]. AB - Actinomycotic infections of the female genital tract are rare. In relation of clinical symptoms and therapeutical implications, reactive and non-reactive infections can be distinguished. During a period of seven years we observed five reactive infections. All these cases with tubo-ovarian abscesses (TOA) were seen in women using intrauterine devices (IUD) for 9.8 years on average. 9.1% of all TOA in this period were caused by actinomyces. Ten cases showed a non-reactive infection (colonisation). The frequency was 1.9%. In three cases pseudosulfurgranules (pseudoactinomycotic granules) were identified histologically by PAS-, Grocott-and Kossa-staining). One case showed a bacterial infection of a mucinous cystadenofibroma of the ovary with colonization of microorganisms similar to cases of botryomycosis in lung and other organs. Reactive infections require antibiotic therapy. Reimplantation of IUD should be done after 4 to 8 weeks, even in areactive infections. In specimens with pseudosulfurgranules no therapy is necessary. PMID- 7483881 TI - [Changes in the vitamin A status in dysplastic epithelium of the cervix]. AB - In 34 patients with HVP-infection of the cervix and in 40 patients with CIN III standardised biopsies were taken from the involved area and normal cervical epithelium for determination of the local concentration of retinylester. In all cases diagnosis was confirmed colposcopically, cytologically and by histology. HPV infection was confirmed by in situ hybridisation. Determination of retinylester was performed by HPLC. No significant difference of local retinyl plamitate concentration was detectable in HPV infected versus normal tissue. Retinyl-palmitate concentration was extremely lower in CIN III compared with normal cervical epithelium and HPV-infected tissue. The determination of plasma level of retinol showed no significant difference between the two groups. So it can be presumed that the reduction of retinyl-palmitat in CIN III is a local process and a local supplementation of Vitamin A might contribute to the prevention of cervical neoplasia. PMID- 7483882 TI - A comparison of survival and side effects in two periods with a different approach to radical hysterectomy as treatment of cervical cancer stages Ib and IIa. AB - The study compares survival and complications during two periods with a different approach to radical hysterectomy as treatment of cervical cancer stages Ib and IIa. Group A included 128 patients operated during the years 1983-87. In that period 5 of the patients who were offered radical hysterectomy had cervical cancer stage IIa. Group B included 135 patients operated during the years 1988 91. In that period 7 of the patients who were offered radical hysterectomy had cervical cancer stage IIa. Considering stage 1b separately, the frequency of operation was raised from 52 (123/237) to 87% (128/148). A 5-years crude survival rate of 85% and 88% was observed in the two groups. The mortality rate was zero in both periods and no fistulae occurred. There were no significant increase in morbidity or length of hospitalization. In the elderly patients over sixty years a significant increase in minor postoperative complications and hospitalization more than 2 weeks were seen. The conclusion is that the frequency of radical hysterectomy as treatment of cervical cancer stage Ib can be raised from 52% to 87% without any noticeable influence on survival or complication rate. PMID- 7483883 TI - [Primary radiotherapy of endometrial carcinoma]. AB - From 1987 to 1993 21 patients with inoperable endometrial carcinoma of all stages received primary radiation therapy. Patients were treated either with combined external beam radiation (EBRT) and afterloading therapy (AL) or AL alone. AL was delivered in high dose rate (HDR) mode. According to tumor stage 3 to 6 times 7 or 10 Gy were applied to line A, one session per week. Due to changing dose prescriptions total doses for EBRT ranged from 45 to 50.4 Gy. 65% of patients died until the end of follow-up (range 6 to 66 months). 23% died due to progressing tumor or therapeutic sequelae (hemorrhagic colitis), 42% died from intercurrent disease. Only 6 patients are alive, one of them with tumor. After a complete course of radiation therapy there was no in-field relapse. One patient irradiated with curative intention developed paraaortal lymph node metastases. During the observation time of our study most patients died of no tumor related reasons. Primary radiation therapy does effectively control endometrial carcinoma. However, old age, reduced general condition and concomitant disease compromise survival rates after radiation therapy alone. PMID- 7483886 TI - [Abdominal actinomycosis in tubo-ovarian abscess]. AB - Diagnosis of actinomycosis as a specific infection is rather difficult. By a case report it is described that preoperative diagnostics and intraoperative situs do not always exactly clarify the situation. In most cases the definite diagnosis can only be achieved by histological findings followed by an adequate therapy. PMID- 7483885 TI - Crohn's disease presented by episiotomy dehiscence and perineal fistula. AB - A case of Crohn's disease is described which was first manifested by episiotomy dehiscence and perineal fistula after an instrumental delivery. The female newborn showed a congenital rectovaginal fistula and hexadactylia. Differential diagnosis and possible relationship between both fistulas are discussed. PMID- 7483884 TI - [Laparoscopic ligation of the internal iliac artery in bleeding cervix carcinoma]. AB - Laparoscopic ligation of internal iliac artery was performed in two patients with bleeding from stage II cervical cancer. The hemostatic effect, shortening of convalescence and early initiation of radiotherapy was obtained. Additionally, during procedure the lymph node sampling was performed. Laparoscopy appeared to be an alternative procedure to abdominal or extraperitoneal ligation of internal iliac artery. PMID- 7483887 TI - Coecal volvulus following caesarean section. AB - A case of coecal volvulus following caesarean section is reported. The importance of knowing the possibility of intestinal obstruction during pregnancy and puerperium is underlined. PMID- 7483888 TI - [Dysphagia: a neurosurgical indication for an operation?]. AB - Ventral osteophytes of the cervical spine can be the reason for dysphagia in rare cases. We present a 67-year old man with difficulties in swallowing for three years and a ventral osteophyte at the level of C 5/6. Native X-rays of the cervical spine, CT-scan and x-ray examination with barium meal and with a barium ball was performed. In our case the most useful procedure for operative indication was the dynamic examination with the barium ball which demonstrates clearly the obstructed oesophageal passage. During operation the ventral osteophyte was resected via ventral approach to the cervical spine. Postoperatively patient's symptoms completely disappeared. Differential diagnosis and therapy are discussed. PMID- 7483889 TI - [Dura-plasty in intracranial operations]. AB - Due to recent reports of slow-virus infections related to the use of lyophilized homologous grafts for duraplasty, we have reviewed the concept of dura substitution in cranial neurosurgery. The present study is based upon 2,115 cranial operations from 1991 to 1994. Frequency and complications of different grafts were investigated retrospectively. Within this time period, dura substitution was required with a frequency of 23.4%. In most cases, autologous tissue was used (47.5%), followed by lyophilized or dehydrated and gamma-radiated homologous cadaveric dura and fascia lata (41.8%). In addition, a synthetic polyesterurethane fleece was used since April 1994. In transsphenoidal approaches to the pituitary gland, autologous fascia lata with or without muscle was used exclusively (100%). In frontobasal trauma, plastic covering of the frontal skull base was performed predominantly by autologous galea-periost (84.8%). In contrast, homologous tissues were preferred for duraplasty in the posterior cranial fossa (77.9%). Complications, including CSF fistula, inflammation, pneumatocephalus, and pseudocysts were observed in 13.1% of operations with dura substitution. The rate of complications was highest in infratentorial (20.0%) and frontobasal operations (18.8%). Application of synthetic grafts for 9 months has reduced complications (6.3%). In conclusion, duraplasty should be generally performed by implantation of autologous grafts such as galea-periost or fascia lata. No restriction of this principle should be accepted for surgery in inflammation, after open brain injury, in the frontobasal region and in transsphenoidal approaches to the pituitary gland. In all other cases, insertion of synthetic grafts seems to be equal to the use of autologous tissue, and even superior in large dural defects.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7483890 TI - Ultrastructural study of cerebral cortex and subcortical white matter following ligation of bridging veins in cats. AB - In 15 adult cats the bridging draining into the superior sagittal sinus as well as their main tributaries were dissected under surgical microscope and ligated. Samples of tissue from cerebral cortex (CC) and subcortical white matter (SWM) with presumed disturbances of venous outflow were investigated by means of electron microscopy following different survival times (ranging from 3 to 15 days). The examination of the ultrathin sections from the CC and SWM disclosed signs of severe oedema: extensive areas of oedema fluid accumulation, swollen dendrited and synaptic endings, neurones with mitochondria destruction, disorganised endoplasmic reticulum and polysomal aggregates, prominent number of vacuoles, dendrites undergoing correspondent changes, degenerating axons and synaptic endings. The conspicuous presence of erythrocytes usually in perivascular zones of the neuropil, occluded, ruptured or collapsed capillaries having vacuoles in their endothelial cells were frequently encountered. These findings confirm the light microscopial observations demonstrating haemorrhagic infarctions after venous occlusions. They provide ultrastructural details about the CC and SWM changes resulting from venous hypertension and hypoperfusion. PMID- 7483891 TI - Neurophysiological techniques in the evaluation of disorders of the spine. AB - Neurophysiological investigations are indispensable tools in evaluating and quantifying functional impairment of spinal cord structures and adjacent nervous tissue in any disorder of the spine accompanied by neurological symptoms. In this review the relevant neurophysiological techniques will be briefly described and their differential value for diagnosis and prognosis of spinal cord damage discussed. Typical case reports of spine disorders at different spinal levels are presented to illustrate results and interpretations of neurophysiological investigations. PMID- 7483892 TI - [Insurance law aspects in expert assessment of isolated traumatic intervertebral disk displacements]. AB - The present paper illustrates the insurance-law aspects of isolated traumatic disc herniation in Germany. The significance of the different terms of insurance in both the personal and the social accident insurance for medical expert opinion is elucidated. PMID- 7483893 TI - [Wilhelm Bruggemann--a neurosurgeon in the Ruhr district (1925-1990)]. AB - Wilhelm Josef Bruggemann, born January 25, 1925, established Neurosurgery in the City of Gelsenkirchen, city of 300,000 inhabitants in the industrial area of the Ruhr District of Germany. After his neurosurgical training with W. Tonnis in Cologne, he worked as Neurosurgeons at the Knappschafts-Hospital "Bergmannsheil" in Gelsenkirchen-Buer from 1967 to 1983. His life and work are commemorated. He died from vascular disease on April 1, 1990. PMID- 7483894 TI - [Modified Kirschner wire insertion for direct ventral screw fixation of dens fractures]. AB - Since 1985 25 patients with a fracture of the odontoid process (dens) were operated in the Neurosurgical Department of the Justus-Liebig-University in Giessen. An early stabilization through an anterior approach was favoured for type II fractures, and for displaced type III injuries, according to the classification of Anderson and d'Alonzo. Two double threaded screws were implanted to reach a compression between the fragments and a stabilization of rotation. With patients having a large thoracic volume and/or a short neck there are often problems, to insert the K-wires in the correct angle, before the screw fixation is performed. For these cases we have developed a new device for the K wire insertion. After technical examinations in specimen the device was successfully applied in clinical use. This device with its inherent advantages is presented and discussed in this paper. PMID- 7483895 TI - A study of two mutant strains of pseudorabies virus (PRV) unable to express thymidine kinase (TK) function. AB - Two mutant strains of pseudorabies virus (PRV) were selected from the virulent 86/27V virus treated with chemical drugs. The viruses, named 6A1 and 6C2, respectively, appeared to be unable to express thymidine kinase function, as demonstrated by the autoradiography test. They showed a reduced virulence for some susceptible animal species (chickens, mice, rabbits, calves, lambs and piglets) and virus was isolated sporadically. The mutant viruses appeared to be able to protect animals against infection with the virulent strain of PRV. At gross, as well as at histological examination, no lesions in apparatus, system and tissues were detected in pigs inoculated with 6A1 and 6C2 viruses. By contrast, rabbits treated with 6C2 mutant strain presented lymphomononucleated cuffs, microgliosis, and neuronophagia in some areas of the brain. This focal spreading, together with the absence of neuronal necrosis and intranuclear inclusions, suggest an infection induced by a modified strain of PRV. PMID- 7483896 TI - Effects of levamisole, DTC and low-dose mechlorethamine on humoral response of SRBC-immunized rabbits exposed to cold stress. AB - The experiments were carried out on normothermal rabbits and rabbits exposed to cold stress (hypothermia). The animals of the latter group were submerged in ice water for 20 s and then placed in a freezer at -15 degrees C for 8 min until their body temperature dropped by 3 degrees C. Both the normothermal and hypothermal rabbits were immunized i.p. with 3 ml of 10% sheep red blood cells (SRBC). Levamisole (2 mg/kg), DTC (sodium diethyldithiocarbamate, 20 mg/kg) or mechlorethamine (mustine; 5 micrograms/kg) were injected i.v. three times at 24-h intervals. The number of PFC, total (19S + 7S) and 2-mercapthoethanol resistant (7S) serum haemagglutination titres were determined. It was found that, in normothermal rabbits, all three agents potentiated the number of plaque-forming cells (PFC); the impact of DTC was the strongest, while the weakest influence was observed for mechlorethamine. Furthermore, DTC increased anti-SRBC haemagglutinin titre, whereas mechlorethamine did not. Levamisole, on the other hand, reduced total serum haemagglutinin titre. Cold stress reduced humoral response to SRBC, which was reflected in the decreased number of PFC and serum haemagglutination titres (19S + 7S and 7S). Each agent showed a different way of action. Pretreatment with DTC prevented the immunosuppression caused by cold stress, while levamisole and mechlorethamine only reduced the immunosuppressive effect. PMID- 7483897 TI - Production of Brucella allergens and evaluation of their biological activity in a guinea-pig bio-assay. AB - A study was conducted to evaluate the biological activity of Brucella allergens extracted with hydrochloride or trichloroacetic acid. Smooth and mucoid Brucella abortus cells and the medium in which brucellae were propagated were used to prepare the allergens. The biological activity of the allergens was estimated in guinea-pigs sensitized with Brucella abortus strain 544. The guinea-pigs were intradermally injected with several allergen dilutions. The dilutions were coded and randomized for site of injection so that none of the dilution was injected twice on the same site. Variance analysis using incomplete Latin square was used for the statistical calculation of the results. The calculated biological activity of the allergens was compared with the biological activity of a 'standard' allergen that has proved effective in detecting cattle brucellosis. The skin erythema diameter was best when recorded 32 h after allergen injection. Statistical analysis of the skin erythema diameters showed a great variation in biological activity (12-105%) between the allergens. Only the allergen extracted from the medium in which a mucoid Brucella strain was propagated was as potent as the standard. The use of the incomplete Latin square for variance analysis resulted in the estimation of the biological activity of nine batches of allergen in only 27 guinea-pigs. PMID- 7483898 TI - Effect of storage on the prevalent alum-precipitated hemorrhagic septicaemia vaccine in Pakistan and preparation of a more efficient oil adjuvant vaccine using dense culture of Pasteurella multocida Roberts type 1 on an improved culture medium. AB - Significantly drastic effects of storage on the potency of the alum-precipitated haemorrhagic septicaemia (APHS) vaccine are reported. The APHS vaccine, studied through challenge infection of vaccinated rabbits (post-60 days of vaccination), showed 100% potency when stored at 4 degrees C for 30 days. The potency dropped to 20% when storage period was extended to 60 or more days. At 30 degrees C, the potency reduced by 40, 40 and 60%, respectively, after 30, 60 and 90 days of storage, while, at 37 degrees C, the decrease was 60, 60 and 100% after 30, 60 and 90 days of storage, respectively. In view of this, the oil-adjuvant (OA) HS vaccine was developed by culturing Pasteurella multocida on a medium comprising yeast extract, sucrose, trypticase and sodium bicarbonate, under continuous aeration at 37 degrees C. This gave a far better bacterial count (maximum count 15 x 10(8)/ml) than the conventional APHS vaccine (maximum count 6 x 10(8)/ml). The OAHS vaccine-carrying water-in-oil emulsion remained stable at room temperature for 1 year. The log protection values of the two batches of the OAHS vaccine, studied in mice, were 5.2 and 5.3, as against 1.9 of the APHS vaccine. PMID- 7483899 TI - Evaluation of immunogenicity of Campylobacter strains isolated from ovine abortions by laboratory test systems. AB - A microagglutination test and in vitro serum bactericidal assay were used for testing cross-reactions and cross-bactericidal activities of 15 Campylobacter strains isolated from ovine abortions. Antisera against certain strains of C. fetus subsp. fetus and C. jejuni showed higher microagglutination titres and bactericidal activity against homologous and/or heterologous strains. In both tests, cross-reactions and bactericidal activity were higher among the strains of homologous species than among those of heterologous species. The pregnant guinea pig model was used for testing the efficacy of experimental campylobacter vaccines. Monovalent bacterins prepared with either C. fetus subsp. fetus or C. jejuni did not produce immunity to cross-challenge with the opposite species. Guinea-pigs immunized with the bivalent bacterin were found to be immune to both groups of Campylobacter when subsequently challenged. In vaccinated guinea-pigs, their in vivo ability to overcome infection is reflected in the in vitro bactericidal capacity of homologous rabbit antiserum. PMID- 7483900 TI - Characteristic properties of streptococci of serological group L. AB - This study was designed to characterize further 88 streptococci of serological group L, isolated from bovines, pigs, poultry and humans. Most group L streptococci from bovines grew as granular sediment with long chains in fluid media and with compact colonies in soft agar. Most group L streptococci from pigs grew with a uniform turbidity in fluid media, short chains, and with diffuse colonies in soft agar. The biochemical characteristics, determined with the Strepzym identification system, revealed no significant differences between the group L streptococci of various origins. A positive opacity factor reaction could mainly be observed with group L streptococci from bovines and humans, but was less pronounced with group L streptococci from pigs and poultry. In addition, 27 group L streptococci reacted with type antigen X or R and 9 cultures with M1 or M6 specific antiserum. The determination of antibiotic resistance patterns revealed that all group L streptococci were resistant to tetracycline and minocycline, part of the cultures were resistant to chloramphenicol, erythromycin and gentamicin, and all cultures were susceptible to penicillin, bacitracin, cefoxitin and nitrofurantoin. All these data allowed an individual characterization of group L streptococci, possibly useful for epidemiological studies. PMID- 7483902 TI - Isolation of the major outer-membrane protein of Actinobacillus pleuropneumoniae and Haemophilus parasuis. AB - A polyclonal antibody against the 35 kDa major outer-membrane protein of Pasteurella multocida cross-reacted with the 40 kDa major outer-membrane protein of Actinobacillus pleuropneumoniae and the 42 kDa major outer-membrane protein of Haemophilus parasuis. The N-terminal amino-acid sequences of these proteins revealed a strong homology with the putative 35 kDa porin protein of Pasteurella multocida (66.7 and 76.2%, respectively). Significant homologies were also evident between the 40 kDa and the 42 kDa protein (76.2%), and with non-specific porins of gram-negative bacteria. PMID- 7483901 TI - Diagnosis of Clostridium perfringens type C enteritis in pigs using a DNA amplification technique (PCR). AB - Clostridium perfringens type C, which produces alpha- and beta-toxin, causes severe haemorrhagic and necrotic enteritis in animals and humans. A polymerase chain-reaction (PCR) assay was developed for the specific detection of the genes encoding alpha-, beta-, epsilon- and entertoxin of C. perfringens for rapid typing of C. perfringens strains, and especially for the identification of type C strains. Both the alpha- and beta-toxin genes were detected directly in porcine C. perfringens type C cultures and also in type B and type C collection strains to a sensitivity of 10(3) cells without purification of the DNA. The alpha-toxin gene was detected in all types of C. perfringens. The epsilon-toxin gene was found in type B and type D, and the enterotoxin gene in some type A strains. Nine other species of Clostridium and a variety of intestinal pathogenic bacteria showed no signal for these toxin genes in this PCR assay. The alpha- and beta toxin genes PCR assay were used to identify C. perfringens strains isolated from intestinal contents of 36 necropsied piglets that had suddenly died or died after premonitory signs of diarrhoea. At necropsy, 20 piglets showed necrotizing enteritis (15 acute and 5 chronic cases) and were suspected to have suffered from a C. perfringens type C infection. All of them had C. perfringens which gave a positive PCR signal for alpha- and beta-toxin genes, and, hence, were identified as type C strains. From the 16 other piglets with lesions other than necrotizing enteritis, C. perfringens strains with the alpha-toxin gene, but no beta-toxin gene, were isolated.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7483903 TI - Virus-neutralization domains on the oligomeric (230 kDa) forms of antigen B of herpesvirus of turkeys and Marek's disease virus differ in cross-serotypic activity. AB - Herpesvirus of turkeys (HVT) is frequently used to protect chickens against Marek's disease (MD). The HVT and MDV native antigen B complex shares common epitopes. To determine whether these oligomers present virus-neutralizing domains, monospecific antibodies to the HVT and MDV native 230 kDa oligomers were produced. The monospecific antibody immunopurified from an anti-HVT avian serum neutralized the in vitro infectivity of the oncogenic isolate MDV-B and the vaccine strains CVI988, SB1 and HVT and immunoblotted the 230 kDa oligomers of HVT and CVI988. As a result of the immunofluorescence analysis on infected cells, the monospecific antibody revealed foci of diffuse cytoplasmic immunofluorescence. A second monospecific antibody to the heat-stable 130 kDa monomer of HVT had limited neutralizing activity against HVT and CVI988 only, immunoblotted only the native HVT oligomer, and was not active in immunofluorescence. The monospecific antibody to the MDV-B 230 kDa oligomer neutralized and immunoblotted only the two MDV-1 strains but stained cells infected with MDVs of the three serotypes in immunofluorescence. It is concluded that the cross-protective neutralizing epitopes of HVT are located on heat-labile oligomeric forms of antigen B. PMID- 7483904 TI - Epidemiological aspects of cerebrospinal elaphostrongylosis in small ruminants in northern Norway. AB - Pathology records from the State Veterinary Laboratory for Northern Norway were used to examine the association between the occurrence of cerebrospinal elaphostrongylosis (CSE) in small ruminants and summer temperature and rainfall in northern Norway. From 1968 to 1993, Elaphostrongylus rangiferi induced CSE was diagnosed in six sheep and 30 dairy goats from 28 different herds in northern Norway. The animals were received for autopsy during 11 different winters. The occurrence of CSE was associated with high mean temperatures during the preceding summers. Further information on clinical CSE and potential risk factors of the disease was obtained from small-ruminant farmers in northern Norway in 1989 and 1991, using a questionnaire. Clinical CSE was defined as posterior paresis without any evidence of brain disturbances or cranial nerve dysfunctions. The survey showed that epidemics of CSE may occur in goat herds while the occurrence of CSE was sporadic in sheep herds. Disease incidence in affected goat herds varied between 0.8 and 8.2%. CSE was associated with the presence of reindeer in the pasture area and occurred in the first, third and, especially, the fourth quarter of the year. PMID- 7483906 TI - Brain lesions in calves following transplacental infection with bovine-virus diarrhoea virus. AB - In 33 calves and subadult cattle of the Holstein-Friesian breed ranging from 1 to 210 days of age, the spectrum of brain lesions induced by intra-uterine infection with bovine-virus diarrhoea virus (BVDV) was retrospectively analysed. Of these, 27 animals originated from herds with a long history of BVD. Six calves were derived from dams vaccinated between the 90th and 118th day of gestation with a BVD live vaccine. The most frequent lesion was cerebellar hypoplasia, being present in 25 out of 33 (76%) of the animals. In most of these cases, cerebellar hypoplasia was associated with hydranencephaly, internal hydrocephalus, microencephaly or porencephaly. In cases with hydranencephaly, the fluid-filled cavities were devoid of ependymal lining. The lumina of the lateral ventricles of these cases were surrounded by glial fibrillary acidic protein (GFAP)-positive cells and a dense layer of immunoreactive cell processes. In the white matter adjacent to the dilated ventricular lumina, a reactive astrocytosis was present. Porencephalic cysts were surrounded by astrocytes with increased expression of GFAP and vimentin-positive cells and cell processes. In hydranencephalic brains, staining for neuron-specific enolase (NSE) revealed a marked reduction of NSE positive nerve cells in cortical areas. In all six experimental cases and in several field cases with hydranencephaly or internal hydrocephalus, small groups of heterotopic NSE-positive neurons were present in the white matter of the cerebral hemispheres. In markedly hypoplastic cerebella, reduction of the cortical cell layers and degenerative changes in, and heterotopia of, Purkinje cells were found. In these cases, NSE- and neurofilament-positive cell processes were either markedly diminished or only remnants of immunoreactive cell processes were present. In five animals without significant gross cerebellar abnormalities, degenerative changes of Purkinje cells were found. Immunohistochemical staining using antibodies against glial and neuron-specific proteins on these brains, which represent postnatal end-stage lesions of BVDV-induced disturbances of the normal brain development, did not provide any insight into the possible pathogenetic mechanisms of these alterations. Application of immunohistochemistry, however, revealed changes, such as reactive astrocytosis and loss of nerve cell processes, which were not obvious on haematoxylin and eosin (H&E) stained sections. PMID- 7483905 TI - Effects of Staphylococcus aureus mastitis after endotoxin application on milk yield and composition during subsequent lactation of guinea-pigs. AB - The effects of Staphylococcus aureus mastitis on milk yield and composition throughout subsequent lactation in lactating guinea-pigs and the role of endotoxin pretreatment on these phenomena were investigated. Primiparous lactating guinea-pigs were intramammarily inoculated with sterile saline (group 1), S. aureus strain UC 6097 (group 2), or with S. aureus UC 6097 after endotoxin pretreatment (group 3). Clinical signs and survival rate were monitored. During the second lactation, daily milk yield was measured and milk composition was determined. In primiparous inoculated guinea-pigs, moderate (group 3)-to-severe acute mastitis (group 2) was produced. During subsequent lactation, milk yield in the control group peaked on day 5 and then decreased. Concentrations of Na+ and Cl- in milk, and concentrations of fat, gradually increased, but lactose and K+ decreased. After an early decrease, NAGase in milk increased towards the end of lactation. Except for higher NAGase concentrations in group 3, milk yield and composition during the second lactation did not differ significantly between the mastitis and the control groups. Endotoxin pretreatment only plays a role in the determination of the severity of the infection. PMID- 7483907 TI - Distribution of serotype, virulence markers and further characteristics of Streptococcus suis isolates from pigs. AB - This study was designed to comparatively investigate 28 S. suis cultures isolated from various pathological processes of pigs. All cultures could be identified biochemically and most of the cultures could be serogrouped with specific antiserum against Lancefield's serogroup D. Serotyping of the S. suis isolates mainly revealed capsular types 2, 1, and 22. In addition, part of the cultures, mostly those of serotype 2, reacted with monoclonal antibodies to the virulence protein muraminidase-released protein, and, in parallel with monoclonal antibodies against the virulence protein extracellular factor. Independently of serotype and the occurrence of both virulence proteins, four cultures haemagglutinated erythrocytes from pigs, humans and rabbits. The haemagglutination reaction appeared to be related to the surface hydrophobicity of the isolates. However, part of the cultures with hydrophobic surfaces did not haemagglutinate the available erythrocyte preparations. The surface characteristics of the S. suis isolates shown in this investigation could be used to characterize individual isolates of S. suis that might be of importance for epidemiological studies. PMID- 7483908 TI - Studies of the detection of Listeria monocytogenes by culture and PCR in cerebrospinal fluid samples from ruminants with listeric encephalitis. AB - A total of 14 cerebrospinal fluid (CSF) samples from ruminants clinically suspected of suffering from listeric encephalitis were examined by polymerase chain reaction (PCR) for the detection of Listeria monocytogenes (L. m.). Of these samples, 11 were examined bacteriologically. Although the clinical diagnosis was confirmed in eight of 11 ruminants by histological and/or bacteriological examination of the brains, L. m. was only detected in one of the CSF samples using PCR, and in none by culture. The PCR-positive CSF sample was obtained from a sheep which had been treated with antibiotics prior to CSF sampling. From these findings, it was concluded that L. m. only occasionally gains access to the meningoventricular system in the course of listeric encephalitis of ruminants and that a reliable aetiological in vivo diagnosis of listeric encephalitis generally cannot be based on the detection of L. m. in the CSF of affected ruminants. PMID- 7483909 TI - [Development of humoral precipitating antibodies to Campylobacter spp. in chickens]. AB - Development of humoral precipitating antibodies against Campylobacter spp. in chickens. The development of precipitating antibodies in chickens was examined by two-dimensional immunodiffusion test after immunization with a formol inactivated vaccine and after subcutaneous and oral application of different live campylobacter serovars. The supernatant of bacterial cells after sonication and centrifugation was used as an antigen in the agar-gel precipitin test. Antisera against different campylobacter serovars showed a high percentage of cross reactions. In chickens immunized with an inactivated vaccine at an age of 1, 2, 3, 4 and 7 weeks, precipitating antibodies could be demonstrated for the first time at 7 days p.i. Except for 1-week-old birds, sera from the other groups reacted positively at 14 days p.o. After subcutaneous duplication of live organisms to 4-week-old chickens, antibodies could already be demonstrated at 4 days p.i. later in part of the experimental groups. No interrelation could be detected between antibody titers, measured by enzyme-linked immunoabsorbent assay (ELISA), from precipitating sera, as well as from those from non-precipitating sera. Precipitating antibodies and antibody titers in the ELISA were examined in sera from groups of birds infected at an age of 1, 2, 3, 4 and 7 weeks. During the Campylobacter excretion period, a distinct peak of antibody titers occurred in 1- and 7-week old birds, whereas other groups showed a steady increase in titers. Precipitating antibodies were only found in 1- and 2-week-old chickens. PMID- 7483911 TI - [Potassium ion transport in the erythrocytes of the frog Rana ridibunda]. AB - The K+ transport in isolated erythrocytes from the frog Rana ridibunda has been studied using 86Rb as a tracer at the temperature of 18-20 degrees C. At physiological K+ concentration (3 mM) in C1- medium ouabain and furosemide inhibited K+ influx by approximately 42 and 47%, respectively. Furosemide had no effect on the Na+ (22Na) transport under the same conditions. The replacement of C1- by NO-3 in medium resulted in significant decrease of total K+ influx, which was not inhibited by furosemide. The ouabain- and furosemide-sensitive components of K+ influx hyperbolically increased as a function of external K+e concentration (1-90 mM) in C1- medium and calculated values of Vmax were 2.2 +/- 0.14 and 5.8 +/- 1.2 mmol/1 cells/h, respectively. K+ influx into frog erythrocytes in the presence of both ouabain and furosemide in NO-3 medium was significantly lower compared with C1- medium. C1--dependent, furosemide-insensitive component of K+ influx increased in a saturable fashion in the range of K+e concentration from 1 to 90 mM. Residual component of K+ transport in NO3 medium in the presence of the blockers was a linear function of K+e concentration and is characterized by the constant of K+ influx rate equal to 0.028 +/- 0.002 h-1. The data obtained indicate that in parallel with the Na, K-pump specific K, C1-transporter participates in K+-transport in the frog erythrocyte membrane. The latter mechanism was only partially blocked by 1 mM furosemide. PMID- 7483910 TI - [A theoretical analysis of the secondary structure and topology of dolichol coupled enzymes]. AB - Hydropathic profile for several dolichol-coupled enzymes (mammal N acetylglucosamine-1-phosphate transferases (GPT), yeast products of genes ALG7, ALG1, DMP1 and SEC59) taking part in the biosynthesis of complex oligosaccharide used for N-glycosylation of proteins in endoplasmic reticulum (ER) of eukaryotic cells has been calculated and constructed. On the basis of analysis of present research and available data sites of amino acid sequence (AAS) potentially capable to penetrate ER membrane were identified. A tendency of AAS segments of dolichol-coupled enzymes to form alpha-helices, beta-folded structures and beta bends was calculated using correlational methods. For COOH-terminal part of ALG1 the prediction about the capability to form helix-helix structures was made. It was concluded that at least three types of topological models in ER membrane were present for the mentioned dolichol-coupled enzymes. PMID- 7483912 TI - [The N-cholinergic activation of the periodic activity of excitable structures in early postnatal ontogeny]. AB - The influence of central N-cholinomimetics on the nature of respiratory rhythm has been studied on 0-16 day rats. It was established that lobeline injection (50 mg/kg) in the rats caused the replacement of background near-second respiratory rhythm by periodic respiration with the period of initiation of respiratory movement complexes of the order of 10-15 sec. Nicotine injection (5 mg/kg) caused less marked effect. On the basis of the present research and available data (the influence of N-cholinomimetics on the spontaneous motor activity and heart rhythm) in was inferred the direct participation of central N-cholinergic structures in reproduction of primary excitement rhythms including the rhythms with the period of 1 and 5-15 sec. The hypothesis on the unity of mechanisms forming the basis of generation of periodic excitement in different excitable structures has been put forward. It was proposed that cholinergic transmitter systems took part in the generation of primary rhythms. PMID- 7483914 TI - [The receptor components of sensory systems: the conservatism of the principles of their interaction]. PMID- 7483915 TI - [The physiology and genesis of hibernation]. PMID- 7483916 TI - [The ciliary motility characteristics of the olfactory receptors in the toad Bufo bufo]. PMID- 7483913 TI - [The participation of opioids in the antinociceptive effect induced by the stimulation of hypothalamic "pleasure zones" in 20- to 30-day-old rabbits]. AB - The involvement of opioid peptides in the mediation of inhibitory influences of the positive reinforcement system ("reward areas") on the evoked potential (EPs) recorded in the thalamic parafascicular complex (CM-Pf) in response to the nociceptive electrodermal stimulation of the hind paw in 20-30 day rabbits has been studied. Electrical stimulation of "reward areas" identified in the preliminary behavioural experiments inhibited the EPs. The systemic injection of naloxone potentiated the antinociceptive effect of "reward areas" stimulation in 75% of cases under the incomplete inhibition of EPs (the decrease of EPs amplitude) but decreased its inhibitory effect in 87% of cases under the complete inhibition of EPs. Naloxone did not change the inhibitory effect of "reward areas" stimulation suggesting to the relationship of analgesic "reward areas" function with nonopioid mechanisms too. On the basis of the comparison of the present data and the ones obtained before it can be inferred that opioid mechanism in realization of antinociceptive influence of "reward areas" stimulation on EPs in CM-Pf is more important for 20-30 day rabbits compared with 41-60 day ones. The correlation of the phenomenon under investigation with the formation of neurotransmitter systems has been discussed. PMID- 7483917 TI - [Lateralization characteristics in the perception of speech in noise by children of different age groups]. AB - The comparative investigation in ontogenesis of the noise immunity of child sensorial perception of two kinds of bioacoustical information (the emotional one being more evolutionary old and the semantic one being evolutionary younger) has shown that the perception effectiveness of these kinds of information depends on the noise level, the age of a child and the kind of information. The perception of semantic information is not provided with the reliable mechanisms of signal identification from noise even to the age of 16 while such mechanisms for perception of emotional information are formed in ontogenesis by the age of 14 and they are associated with the activity of the right hemisphere. PMID- 7483918 TI - [A comparative analysis of the changes in the sleep phases of rats genetically predisposed to audiogenic seizure attacks]. AB - The parameters of diurnal wakefulness-sleep cycle in norm for Wistar rats with low sensitivity to convulsive sound action and KM rats with genetic predisposition to audiogenic seizures are presented. The analysis of the nature of changes in cycle structure for KM rats after short audiogenic generalized paroxysmal seizure is given. PMID- 7483919 TI - [The search for homologous sequences in the primary structure of dolichol-coupled enzymes]. AB - Primary structures of several dolichol-coupled enzymes, mammal and yeast N acetylglucosaminylphosphote transferases (GPT), yeast mannosyl transferase and dolicholphosphomannosyl transferase were compared. The enzymes presented take part in the synthesis of oligosaccharide precursor used for N-glycosylation of proteins in endoplasmic reticulum of eukaryotic cells. For GPT family equalization of amino acid sequences (AAS) was conducted and the profile of invariability of the primary structure was calculated. At the comparison of AAS in the enzymes outlined with the ones in carbohydrate-binding proteins homologous segments were identified and their possible role was discussed. PMID- 7483920 TI - [The effect of host temperature on the fatty acid composition of the total and membrane lipids in the cestodes Eubothrium crassum and Diphyllobothrium dendriticum]. AB - A comparative qualitative and quantitative analysis of the fatty acid composition of total and lysosome membrane lipids of two cestode species Eubothrium crassum (from cold-blooded host) and Diphyllobothrium dendriticum (from warm-blooded host) has been carried out. It is shown that quantitative ratio of the fatty acids in the total and membranous lipids of the helminth of cold-blooded host was very similar whereas for the helminth of warm-blooded host fatty acid composition of membranous lipids was considerably more saturated compared with the one of total lipids. It is assumed that the high level of unsaturation of total lipids from D. dendriticum is associated with preadaptation mechanism of fatty acid spectra in parasites with the development coupled with a consecutive rapid change of different thermal regime habitats. PMID- 7483922 TI - [The development of the active sleep phase in phylogeny and ontogeny]. PMID- 7483921 TI - [The temporal characteristics of "winter" paradoxical sleep in the hibernating suslik Citellus major]. AB - It has been shown that paradoxical sleep (PS) total time (TT) in euthermia phase of hibernation cycle (winter sleep) can be compared with its mean values in euthermia season. Ultradian fluctuations of PS episodes duration, TT and phase number as well as bradycardia microintervals are revealed in euthermia phase dynamics. Amplitude of these fluctuations increases before hibernation entry; in transitional "start" period PSTT increases up to maximum critical values. In hibernation cycle PSTT rapidly decreases at the first stage owing to reduction of PS phase number and their duration. These data support and supplement earlier suggested hypothesis about endogenous "swinging up" of ultradian rhythms of sleep and thermoregulation promoting stage-to-stage expansion of homeostasis temperature limits in various critical periods of preparation and entry of homeotherms into natural hypometabolic states (torpor, hibernation and others). PMID- 7483923 TI - [A comparative analysis of the oxygen-binding and antioxidant properties of the blood of laboratory animals and the muskrat Ondatra zibethica]. AB - Close correlation of oxygen-binding properties of haemoglobin and the content of malonyl dialdehyde (MDA) in animal blood plasma was shown. The higher the affinity of haemoglobin to O2, the lower MDA level in the blood plasma. For the muskrat Ondatra zibethica, representing diving animals, MDA content is lower and catalase activity is higher compared with the laboratory rat. PMID- 7483924 TI - [The theoretical justifications for the application of the institution of limited responsibility]. PMID- 7483926 TI - [Pseudostroke states in elderly patients]. AB - Clinical characteristics of pseudostroke conditions in connection with the nature of somatic pathology, responsible for those conditions, were investigated in 52 patients. According to the character of somatic pathology all the elder patients were divided into 4 groups: myocardial infarction and progressive heart insufficiency, acute lung and heart insufficiency, inflammatory kidney disorders with signs of renal insufficiency, different intoxications. According to the results of macroscopic brain investigation, there were post-ischaemic cysts. Pathogenetic aspects of post-stroke conditions are discussed. PMID- 7483925 TI - [The clinico-psychological aspects in the decompensation of the initial forms of a cerebral vascular lesion]. AB - Clinico-neurological, instrumental and psychological methods were used to examine 35 patients at the of 55-70 years with early stages of cerebrovascular insufficiency. It was found out that subjective signs of cerebral vascular disorders were likely to be due to nonspecific cerebral dysfunction and associated with psychical dysadaptation. The degree of memory and attention disorders negatively correlated with intensity of mental activity. The results of the work may be used in prophylaxis of decompensation of cerebrovascular pathology. PMID- 7483927 TI - [The clinical characteristics of rhinosinusogenic diffuse cerebral arachnoiditis located predominantly in the anterior cranial fossa]. AB - 142 patients with rhinosinusogenic cerebral arachnoiditis (RCA) were investigated. Diagnosis of RCA in all cases was confirmed by results of otorhinolaryngologic and neurologic methods of investigation. Pneumoencephalography was performed in 60 cases. In order to achieve auxiliary contrasting of meninges anterior cranial space the new way of position of patient on the table for X-ray examination were used with a special device. Analysis of pneumoencephalograms of 30 patients, performed with the new position and the new device allowed to detect predominant location and character of meninges pathology in PCA. Diagnostic significance of pneumoencephalography increased because of additional air depot in brain coverings of anterior cranial space. On the basis of clinical and pneumoencephalographic data it was concluded, that in cases with RCA meninges of anterior cranial space are the place of primary location of pathological process. The main topical of RCA was diffuse cerebral arachnoiditis with predominant pathology meninges of anterior cranial space (66.9% of patients). Among the main clinical peculiarities of this form were the following: characteristic of head pain in supraorbital area with the feeling of the pressure on eyes and painful eye movements, frequency of smell and psychic disorders. Analysis of clinical and pneumoencephalographic data demonstrated, that pathological changes in brain meninges and brain matter prevailed at the side of rhinosinusitis or of more damaged sinus in cases with bilateral disease. PMID- 7483928 TI - [The results of using therapeutic periorbital electrostimulations in neurology patients with partial atrophy of the optic nerves]. AB - Results of clinical application of method of non-contact periorbital electrostimulation in 6 groups of neurologic patients (total number-246) with optic nerve pathology of different origin (post-traumatic, hypertensive, inflammatory) are summarized. Analysis of interdependence between positive results and severity of visual disorders, type of the disease, responsible for partial atrophy of optic nerve, was performed. This method of restoration of vision in investigated patients proved to be effective both in hospitals and outpatient departments. PMID- 7483929 TI - [Late complications of poliomyelitis]. AB - We examined 73 31-55-year-old postpoliomyelitis patients who had paralytic form of the disease before the age of 11. It was found that progression of neuro muscular signs (pareses, atrophies, myalgias), typical for postpoliomyelitis muscular atrophy (PMA) emerged in 43 patients. Classical statistical analysis of clinical and electromyographical data and special statistical method of extreme parameter grouping confirmed PMA only in 8 out of the above 43; 3 forms of postpoliomyelitis syndrome were recognised. PMID- 7483930 TI - [Russian preparations for the metabolic therapy of multiple sclerosis patients]. PMID- 7483931 TI - [The epidemiology of vascular brain diseases]. AB - Prevalence of different forms of acute cerebral accidents (ACA) and factors, provoking their development, has been investigated in the population of Kishinev. Annual ACA morbidity made up 226 per 100000. More than 80% of ACA patients had brain infarction (BI), 13.5%--intracerebral and 6.5% subarachnoidal hemorrhage. High BI risk has been established in patients with arterial hypertension, heart failure, elevated blood levels of sugar, cholesterol, beta-lipoprotein as well as with increased blood viscosity. PMID- 7483932 TI - [The pathogenetic mechanisms of the development of experimental allergic polyradiculoneuritis]. AB - Biochemical, immune and morphological changes in peripheral nervous system at different stages of experimental allergic polyradiculoneuritis were studied in 102 rabbits as indicated by total lipids, cholesterol, phospholipids and their fractions, diene conjugates of spinal ganglions, cervical, thoracic and lumbosacral roots, trigeminal ganglions and nerve trunks of extremities. It was found out, that derangement of lipid metabolism in spinal ganglions and nerve roots is the primary phase in the development of the disease. Those could be seen as early as the experiment day 6. From the 8th day after the injection of antigen (myelin in complete Freund adjuvant) impairment of protective function of blood nerve barrier took place in the form of increased vascular permeability of spinal ganglions, roots and trigeminal ganglions. Fixed antibodies, located on myelin sheath predominantly of intraganglion fibres of lumbar, cervical and thoracic segments, trigeminal ganglions, appeared after these changes. Demyelination and inflammatory infiltration began in the end of the incubation against initial clinical signs of polyneuritis in rabbits and reached its maximum at the peak of the disease. PMID- 7483933 TI - [The formation of a collection of DNA from patients with endogenous psychoses and the prospects for its use in molecular genetic research in psychiatry]. AB - A genetic study of psychiatric diseases meets a lot of difficulties because of incomplete penetrance, phenocopies, probable genetic heterogeneity. The progress in exploring of new effective markers and human genome mapping enables one to undertaken a systematic search for alleles confirming susceptibility to manic depressive illness and schizophrenia. It is important therefore to have in one's disposal the collection of DNA samples appropriate for different aspects of molecular genetic study. The collection of DNA samples deposited in National Centre for Mental Health of Russian Academy of Medical Sciences includes DNA of informative small-scaled families and a number of DNA from unrelated subjects with manic-depressive illness, schizophrenia, schizoaffective disorders. This collection may be used for linkage analysis, association and sib-pair studies and for detection of mutations responsible for mental diseases. PMID- 7483936 TI - [Patterns of the syndrome transformations in recurrences of alcoholic psychoses (clinico-statistical research)]. AB - Psychopathological aspects of recurrent attacks of alcoholic psychosis has been investigated. The article presents results of examination of 689 patients with alcoholic psychosis (407 with delirium, 189 with hallucinations, 93-paranoids), who have suffered 2093 psychotic attacks within 10 recurrences. It was found, that change of syndromes from the 1st to the 10th attack followed in a declining order and the character of syndromologic transformation was steadily-advanced. Two variants of transformation of alcoholic psychosis were revealed: the tendency to preservation of an initial clinical form during the following recurrent attacks and the tendency to changing psychopathologic syndrome. PMID- 7483934 TI - [A multivariate genetic analysis of the data from a complex study of the predisposition to schizophrenia]. AB - 49 families of schizophrenics (49 probands and their 106 first-degree relatives) were studied by means of psychological methods, EEG and computed tomography. Multivariate genetic analysis were performed to assess genetic and environmental components of parameter phenotypic variance and relations between the parameters and liability to schizophrenia, including formation of discriminant functions for classification of individuals into high risk and normal groups and estimation of the functions heritability. Besides, correlations between parameters of neuromorphological, neurophysiological and psychological levels were calculated. The heritability of most EEG and CT parameters were between 41 and 98%. Among psychological variables only mediated and logic memory seem to be under sufficient genetic control (relatively 62 and 42%). Discriminant functions derived from psychological and EEG parameters, allow to differentiate correctly individuals into high risk and normal groups in 76-88% cases. Results of correlational analysis of psychological and biological variables suggest that there are both system and separate genetically determined abnormalities related to different levels of pathogenesis. PMID- 7483935 TI - [The follow-up of therapeutic remission in alcoholics]. AB - Usage of alcohol during the period of therapeutic remission of chronic alcoholism without complete restoration of its symptoms indicates failure of therapeutic remission (FTR). A method was suggested to detect FTR by enzymic activity of gamma-glutamyltranspeptidase, aspartate transaminase, alanintransaminase and glutamate dehydrogenase. FTR is stated at differential threshold of the above enzymes 32, 25, 26 and 6.5 u/l, respectively. Validity of the method was confirmed at examination of 110 chronic alcoholics and 61 healthy persons. Early FTR detection prevents occurrence of true recurrence of chronic alcoholism. PMID- 7483938 TI - [Patient opinion about the quality of psychiatric care]. AB - The author has provided a study of patient's satisfaction with the quality of mental care. The opinion of 277 patients who used traditional services (psychoneurological out-patient clinic, psychiatric hospital, day hospital and work-shop) was analyzed. The data that were obtained show that despite a satisfaction with the quality of care on the whole there are essential disadvantages in the organization of psychiatric care. Relying on patient's opinion some practical recommendation for improvement of the activity of mental health services are supplemented. PMID- 7483937 TI - [The clinico-epidemiological characteristics of a contingent of older patients being cared for by the psychoneurological office of a regional polyclinic]. AB - Psychic state of the majority (84.5%) of 410 consulted by the psychiatrist in a district outpatient clinic patients was found as non-psychotic. Psychic disorders of exogenic-organic nature accounted for 34.4% and neurosis--26.5% of all the cases. Psychoneurological care, available in the district outpatient clinic, is an adequate form of aid for patients with borderline psychic disorders. PMID- 7483939 TI - [The estimation of the public danger of schizophrenia patients based on epidemiological research data]. AB - The article presents the proportion of different categories of patients with schizophrenia performing socially dangerous actions (SDA) in the total number of registered patients. It describes features of the disease, sex, age specific for clinical presentation background. Complex evaluation of factors, including premorbid of personality traits and environmental phenomena is considered to be very important because of its prognostic significance. In cases with less acute and less progressive course of the disease significance of social and psychological factors increases. Difficulties in up-to-date detection of patients, inclined to SDA, are analysed. PMID- 7483940 TI - [Ancient philosophy as a source of ideas for "borderline" psychiatry in the historical and epistemological aspect]. PMID- 7483941 TI - [P. I. Iakobii and the creation of outpatient psychiatric care in Russia (on the 150th anniversary of his birth)]. PMID- 7483942 TI - [The ultrasonic diagnosis of the Arnold-Chiari malformation in young infants]. PMID- 7483943 TI - [The prognosis for the development of ischemic disorders of the cerebral circulation in persons with an asymptomatic course of atherosclerotic occlusive lesions of the major arteries of the head]. PMID- 7483945 TI - [Multiple arteriovenous malformations of the brain, spinal cord, kidneys and peripheral vessels]. PMID- 7483944 TI - [A case of the clinical manifestation of the Arnold-Chiari syndrome in middle age]. AB - The article presents description of Arnold-Chiari syndrome in 62-year-old woman. The diagnosis was made clinically, by means of MRT and verified during the operation. PMID- 7483946 TI - [Central nervous system involvement in the acquired immunodeficiency syndrome]. PMID- 7483948 TI - [Rhinological data in the diagnosis and choice of the type of surgical intervention in cerebrospinal rhinorrhea]. AB - The study covers 70 cases of liquorrhea nasalis of various etiology. ENT specialists frequently regard liquorrhea nasalis as vasomotor allergic rhinitis. Liquorrhea nasalis may be divided into craniosinusonasal and cranionasal ones. The latter is more commonly followed by intracranial complications. An otorhinolaryngological otoneurological examination as part of a comprehensive study of patients with liquorrhea nasalis aids in diagnosing the disease and reveals the results of surgical treatment from the nasal endoscopic data, detects intranasal cerebral hernias, liquor cysts and tumors in the nasal cavity, residual otoneurological symptoms after prior meningitides, brain injuries and other diseases. A patient's positions and conditions allowing one to successfully detect even minimum liquorrhea nasalis have been worked out. Liquorrhea nasalis is subdivided into liquorrhea: a) that has no concurrent changes in the nasal cavity; b) that is accompanied by intranasal hernias and liquor cysts; c) that is concomitant with purulent sinusitis. The indications for any surgical intervention are different in each group, the rhinoscopic findings may be decisive in its choice. Among residual symptoms in patients with liquorrhea nasalis there are the most common olfactory disorders (35%) and mainly neurosensory hearing decrease (30%), beginning from mild hearing impairments resulting in complete deafness without vestibular excitability. PMID- 7483949 TI - [The diagnostic and treatment characteristics of birth injury to the brachial plexus in children and adolescents]. AB - From 1980 to 1994, a total of 73 patients were treated at the Russian Research Surgery Institute named after A. L. Polenov for damage to the brachial plexus during birth injury. The authors proposed an original procedure for treating these patients. This involved conservative therapy, surgical intervention on the brachial plexus and correction of the arm locomotor apparatus by orthopedic techniques. The conservative treatment were performed by original methods: namely in combination with hyperbaric oxygenation, low-energy laser exposure of the brachial plexus or by administering cholinesterase agents into the bioenergetic points, followed by electrostimulation of the neuromuscular apparatus. The original conservative treatment led to a significantly improved arm function in 53% of the patients. If the conservative therapy failed, obstacles to the restoring the conduction of the brachial plexus were eliminated via operations (neurolysis, suture, autoplasty, reinnervation). The operation gave the benefit to 74% in whom conservative therapy turned out to be ineffective. If surgical recovery of the brachial plexus failed, arm locomotor intervention was applied. PMID- 7483950 TI - [The results of reconstructive microsurgical operations on the damaged common peroneal nerve and its deep branch in children]. PMID- 7483947 TI - [The use of hyperventilation for improving the visualization of glial tumors of the brain in magnetic resonance tomography using the contrast substance Gd-DTPA]. AB - This study was undertaken to evaluate the influence of hyperventilation (HV) as a test on the image contrast of brain glial tumors in enhanced magnetic resonance (MR) imaging. HV was performed for 2 min (30 breaths/min) before intravenous injection of Gd-DTPA (Magnevist), Schering AG, Germany, in a dose of 0.1 mmol/kg. A total of 19 patients with glial tumors of the brain were examined. After the functional test, the image contrast of tumor tissue was compared with tumor contrast after the standard enhanced MR imaging. Nineteen patients with brain tumors were assessed. The tumor types were histologically verified in all cases (nine malignancy degree I-II astrocytomas, six malignancy degree III astrocytomas, three ependymomas, one malignant oligodendroglioma). MRI was performed on a Magnetom 42 SP-1.0 T. There was an increase in the image contrast of degree I-II astrocytomas neither after the standard enhanced MRI nor after it in the presence of HV. On the contrary, in 2 cases there was an opposite effect- the tumor contrast decrease after the functional test. Ependymomas showed patterns of increases in contrast intensity from 10 to 13% as compared with the enhanced standard MRI in all cases. In these conditions the intrinsic structure and boundaries of tumors became more distinguished. Cases with malignant astrocytomas and oligodendroglioma had contrast enhancement increases as well. It is concluded that in cases with degree I-II astrocytomas, the use of HV does not improve the visualization of lesions. In ependymomas and anaplastic astrocytomas, HV aids in more significantly assessing the intrinsic structure and the extent of tumorigenesis and it may be useful as a functional test to assess the anaplastic extent of glial tumors of the brain and as a procedure enabling the contrast agent to be used in smaller dosages. PMID- 7483951 TI - [The retraction pressure during neurosurgical operations on the brain. III. An analysis of the efficacy of prophylactic methods aimed at reducing brain volume]. AB - The comparison was undertaken to examine 31 patients with various brain neurosurgical abnormalities to evaluate the efficiency of some of the most widely used preventive measures for retraction ischemia, which are aimed at reducing the brain volume at surgery: preoperative administration of a saluretic, osmodiuretic solution, hyperventilation, tumor cyst puncture, lumbar and ventricular drainage. The findings suggest the efficacy of such approaches, as tumor cyst puncture, intravenous administration of an osmodiuretic, ventricular and, possibly, lumbar drainage. Such methods as preadministration of saluretic hyperventilation are ineffective. PMID- 7483952 TI - [The immunological reactions and enzymatic activity of the cerebrospinal fluid in intracranial complications following neurosurgical operations]. AB - The specific features of immunological responses and enzymatic activity of cerebrospinal fluid in the development of intracranial infectious complications were outlined in 16 neurosurgical patients after surgical interventions. On days 1-3-5 days after surgery, the immunobiochemical spectrum of cerebrospinal fluid showed substantial changes associated with the impaired permeability of the blood brain barrier and the stress-induced transition of biochemical functional systems on the minimum functioning during anaerobic energy supply. Increasing lipid peroxidation processes reflect the result of catabolic reactions and energy deficiency, followed by intensive cytolysis of leukocytes and nerve cells. With this, antibody synthesis and great rises in the formation of circulating immune complexes is the essence of sanogenetic mechanisms basically aimed at eliminating the released and bound antigen in the given time interval. PMID- 7483953 TI - [The induction of lipid peroxidation in human brain glial tumors]. AB - Tissue examinations of glial tumors in the human brain revealed that therein lipid peroxidation could be induced by using bivalent iron salts, which is indicated by higher malonic dialdehyde levels. The authors have demonstrated that the glioma tissue levels of iron were statistically lower than those in the brain tissue. The induction in tumor tissue does not depend upon the degree of its malignancy, but it significantly differs from this parameter in the rabbit brain tissue. The induction of lipid peroxidation processes is accompanied by a lower cumulative antioxidative activity. The findings open new prospects for affecting tumor growth. PMID- 7483954 TI - [Experience with using mobile x-ray computed tomography in the diagnosis of hypophyseal adenomas]. PMID- 7483957 TI - [Bilateral neurinomas of the acoustic nerve in children]. PMID- 7483956 TI - [The surgical treatment of an aneurysm with a carotid-ophthalmic location]. PMID- 7483955 TI - [Stereotaxic destruction of the ventrolateral thalamic nucleus in combination with the transplantation of fetal and xenogeneic tissue in Parkinson's disease]. AB - Four patients with Stage III-IV Parkinson's disease were operated on. Stereotactic cryodestruction of the ventrolateral nucleus was made, followed 10 minutes later by implantation of mixed fetal and xenogenic nerve tissue cells. The nerve cells-the fetal part-are obtained from human fetuses in the age range of 14-18 weeks of development. Tissues from the ventral portion of the midbrain and basal ganglia of the forebrain were used. Neurogenic layings of Notch Drosophila melanogaster mutants serve as a xenograft. The mixture of nerve cells were prepared in ratios of 1000:20 and 1000:12. The follow-up of the patients lasted a year. Relapses were absent. The effect occurred on the operating table (in the combination of nuclear destruction and nerve cell transplantation) and only 6 months later only during transplantation of nerve cells (without destruction). An attempt of transplanting the above fetal xenogenic mixture of nerve cells to patients previously undergone 2 ineffective stereotactic operations provided no significant clinical effect. The concurrent destruction of ventrolateral nuclei in combination with fetal xenogenic tissue grafting is considered to be promising. PMID- 7483958 TI - [Microvascular decompression]. PMID- 7483959 TI - [Improved methods for electrocorticography during single-stage and long-term stereotaxic interventions]. PMID- 7483962 TI - Human population growth: local dynamics--global effects. AB - This communication presents a very simple model for the global growth of the human population. It is shown that the solution of the simple equation x = beta x2 describes the experimental data remarkably well. This equation is derived by introducing a new, local concept: the individual quality of life and by assuming that human breeding strategy consists in keeping this quantity constant through the ages. However the model should be enlarged with a saturation mechanism if future growth is to be described. PMID- 7483961 TI - [The clinical classification of the bone defects as a consequence of craniocerebral trauma]. PMID- 7483960 TI - [The x-ray anatomical structure of the anterior portions of the circulus arteriosus (Willisi) cerebri in aneurysms of the anterior cerebral-anterior communicating arteries]. AB - The angiographic findings were analyzed in 136 patients with aneurysms of forebrain and anterior communicating arteries and 27 patients examined for brain extravascular abnormality. Depending on the magnitude of effects of one of the internal carotids on the blood supply anterior communicating and forebrain arteries, symmetric and asymmetric filling variants for the anterior portions of Willis' circle were identified. The symmetric variant was detected in 30% of cases and in 67% of controls, the asymmetric was revealed in 70 and 33%, respectively. One of the ICAs was prevalent in filling the anterior portions of Willis's circle in 70.3% of patients with aneurysms and in 90.6% of cases the diameter of its ipsilateral segment A1 was larger than of the segment contralateral to A1. Aneurysms were located in the trunk of the anterior communicating artery in 13% of cases and in the corner of the forebrain artery and the anterior communicating artery in 87% of cases. Most aneurysms were located on the side of the prevailing segment A1. The influence of a hemodynamic factor on the formation of aneurysms in the area of the anterior communicating and forebrain arteries. PMID- 7483963 TI - Why animals have tumours. AB - From the viewpoint of an evolutionary biologist, carcinogenesis should be looked upon as a protective mechanism against destruction of DNA. Because genes expressed in embryonic cells are covered and protected by heterochromatinization, they are the most appropriate 'alternate genes' compared to genes that are expressed already in somatic cells. When DNA-damage occurs, the embryonic genes can be activated. Some somatic cells exhibit some features of embryonic cells. PMID- 7483964 TI - Concurrent effect of morphine on thalamic nociceptive neurons and medullary on- and off-cells. AB - On- and off-cells of the rostral ventromedial medulla project to the spinal cord, where off-cells are postulated ot inhibit, and on-cells to facilitate, nociceptive transmission. In the present experiments, unitary recordings were made from ventrobasal (VB) thalamic neurons and, simultaneously, on- or off-cells in lightly anesthetized rats. When the tail was heated (45-55 degrees C) off cells stopped firing (pause) and on-cells began to fire (burst) 0.5-0.6s before VB neurons responded to the tail stimulus. Administration of morphine sulfate (2 mg/kg i.v.) concurrently prevented the off-cell pause, the on-cell burst an the VB neuronal response. Naloxone (1 mg/kg i.v.) reversed these effects. The results are consistent with the notion that prevention of the off-cell pause and the on cell burst by morphine is responsible for the lack of nociceptive information flow towards the thalamus. PMID- 7483965 TI - [Genetic and demographic structure of the State of Aragua, Venezuela, estimated through the surnames]. AB - The genetic structure of the State of Aragua, Venezuela, has been studied through the analysis of surnames obtained from the register of electors. The analysis covered 23 counties and included a total of 99,593 individuals and 6,338 different surnames. Estimators of isolation, consanguinity, microdifferentiation and four measures of genetic distance, were studied. When our results were compared with those obtained in other States of Venezuela studied previously (Falcon, Lara, Merida, Nueva Esparta and Yaracuy), Aragua appears as the State most open to new migrants, probably due to its nearness to Caracas, Venezuela's capital city. Within Aragua, the counties most isolated are Choronii, Ocumare de la Costa and Tovar, while those less isolated are El Limon, Turmero, La Victoria, San Mateo and Cagua. The correlations between the logarithmic transformations of genetic and geographic distances were all significant, revealing surname differentiation by distance. The dendrogram built with the Euclidean distance matrix shows a first group of counties formed by those localized towards the central portion of the State, to which southern counties are added. Choroni and Ocumare de la Costa form a group which enters the dendrogram just before Tovar, the last county to be included in the analysis. Seven surnames with a focal distribution within the State were identified: Ayala and Calanche in Choroni; Kanzler, Misle and Ruh in Tovar; Lira in San Mateo and Santaella in Barbacoas. PMID- 7483966 TI - Skeletal muscle ultrastructural and biochemical alterations induced by experimental hyperthyroidism. AB - Rats treated with high doses of T4 showed an increase in the oxygen consumption in state 3 rate respiration of mitochondria isolated from Soleus (slow twitch) and Extensor Digitorum Longus (EDL, fast twitch) muscles, without modification of either their phosphorylation efficiency (ADP/O ratio) or their coupling. Muscle fiber main alterations were atrophy and changes of the structure and shape of mitochondria. Also, abundant autophagic vacuoles with mitochondrial debris, myelin-like figures, glycogenosomes and primary lysosomes were found. PMID- 7483967 TI - [Biodiversity and similarity between the nematode parasite communities of domestic and wild ruminants]. AB - The analysis of the bibliografical information concerning to the nematode parasite communities of domestic and wild ruminants, permitted to obtein the following values of Maximum Diversity Index, Bovidae: 4.88 bits; Cervidae: 4.33 bits and Cameliae: 3.69 bits. The highest values of this index within each family of host wee 5.75 bits. 5.49 bits and 4.32 bits for Ovis aries (Bovidae), Capreolus capreolus (Cervidae), Lama pacos and Camelus dromedarius (Camelidae), respectively. The families of parasites with direct biological cycles, showed higher values of Maximun Diversity Index than those families with parasites that require vectors or intermediates hosts in their biological cycles. Concerning to the Similarity Index among the hosts in relation to their communities of nematode parasite, the highest values (> 60%) were presented between the following species: Ovis aries-Capra hircus; O. aries-Bos taurus; B. taurus-C. hircus; Capra ibex-Rupicapra rupicapra; Capreolus capreolus-Cervus elaphus; C. elaphus-Dama dama; Lama pacos-Vicugna vicugna; D. dama.-C. capreolus. PMID- 7483969 TI - 2-Chloro-2'-deoxyadenosine. PMID- 7483968 TI - [Biomass production enriched in intracellular methionine by a mutant of Saccharomyces cerevisiae]. AB - According with FAO reported data the methionine intracellular content in Saccharomyces cerevisiae is higher than another yeast. For increasing the yeast methionine internal concentration three S. cerevisiae mutant strains were chosen (M2, M4 y M9), obtained by ethionine (0.1 mg/ml) and norleucine (0.33 mg/ml) resistance by Gonzalez Miliani (personal communication). The resistance levels in culture were modified until selection of a mutant LF-M9 etr norr, which shows resistance to ethionine (6 mg/ml). Optimal conditions for growth were fixed on shaked flasks and later mutationaly experiments were conducted with nitrosoguanidine (0.5 mg/ml) and U.V. light (240 nm in 9 minutes d = 32 cm). Mutants obtained were selected on plate replicates and microbiological test, using Escherichia coli 303 (Wollman, met- b1- strr) as the indicating strain. The mutant LF-M9 treo- etr norr, shows an intracellular methionine content 3 times higher than the control strain DSM D273-10B and 1.8 times higher than parental strain M9. The mutant was cultivated on different agroindustrial wastes and the optimal growth was reached in acid hydrolysates of cassava foliage. Fermentations in 1 litre stirred fermentor were accomplished using these medium and the biomass obtained was 6.3 g of the yeast (dry weight) enriched in methionine per litre of extract. PMID- 7483970 TI - [Epidemic of hantavirus disease in Entre-Sambre-et-Meuse: year 1992-1993. Clinical and biological aspects]. AB - A multihospital study allowed us to follow a total of 62 serologically proven cases of Nephropathia epidemica (NE) in the Belgian region between Sambre and Meuse during the 1992-1993 period. The clinical picture consisted of sudden high fever (100% of the cases), headache (71%), abdominal or lumbar pains (80%) and, as a less frequent but very specific sign, acute myopia (24%). Non-specific respiratory symptoms such as a non-productive cough and an abnormal lung auscultation were found in 1 case out of 4. Frequent laboratory anomalies were thrombocytopenia (69%), left-shift leucocytosis (77%), abnormal LDH (69%) and an inflammatory syndrome (86%) with levels of C-reactive protein (CRP) elevated up to a mean of 102 mg/L, often accompanied by a marked fall of total serum cholesterol and a rise of triglycerides. Impaired kidney function is the rule (84%), nevertheless serum creatinine levels remain inferior to 150 mumol/L (1.7 mg%) in 25% of the patients. As for neurological signs, 1 case of encephalitis and 1 syndrome of Guillain-Barre were observed. Relative bradycardia (< 90 bpm) was noted in 50% of the cases with fever, whereas Doppler-echocardiography detected pericarditis in 1 case, and transient impairment of the left ventricle function in 3 cases. Frequent elevation of liver enzymes (46%) confirms the observation that from now on, Hantavirus infections should be considered in the differential diagnosis of viral hepatitis. One severe case was observed with shock, diffuse intravascular coagulation and adult respiratory distress syndrome (ARDS), followed by 5 other patients presenting with marked degrees of hypoxemia and hypocapnia. We conclude that not only the causative Hantaviral serotype, but also the degree of "systemic inflammatory response syndrome" (SIRS) seems to determine the clinical picture in Hantavirus infections. PMID- 7483971 TI - [Systemic sarcoidosis with cutaneous lesions: literature review]. AB - Sarcoidosis is a systemic disease of unknown etiology. The diagnosis is based on consistent clinical, biological and radiological findings supported by histologic evidence of noncaseating epithelioid granulomas. Skin lesions occur in about a quarter of patients with sarcoidosis. The purpose of this article is to analyse through the clinical presentation of dermatological lesions, a potential link with systemic disease. PMID- 7483972 TI - Senile (type II) osteoporosis: pathogenesis and prevention of estrogen deficiency related bone loss at the proximal femur. AB - Fractures of the proximal femur are associated with considerable morbidity and mortality. The residual effects of estrogen deficiency-related bone loss (partially) account for the higher incidence of hip fracture in elderly women compared to elderly men. This accelerated phase of postmenopausal bone loss can be prevented by estrogen substitution therapy. However, when estrogen substitution is discontinued, bone density declines at a rate similar to the perimenopausal one. In line with these densitometric data, several studies have provided evidence that the protective effect of estrogen therapy on hip fracture incidence does not persist after discontinuation of substitution. To prevent hip fractures in old age, estrogen treatment may have to be initiated perimenopausally and never discontinued. At present, however, such a strategy cannot be recommended as it has not been settled yet whether the benefits of estrogen substitution outweigh the risks. PMID- 7483973 TI - Malignant peritoneal mesothelioma. Case report and review of the literature. AB - We describe a peritoneal mesothelioma. There are many aspecific symptoms. Professional exposure is found in only fifty percent of cases. The histological diagnosis is often difficult. The survival period is short because of the absence of curative treatment. PMID- 7483974 TI - Metastatic pleural effusion as presenting sign of prostatic carcinoma. AB - A patient is reported with symptomatic bilateral malignant pleural effusions as the only and presenting sign of prostatic carcinoma. In view of the excellent response with hormonal therapy, this case illustrates the importance of including prostatic carcinoma into the differential diagnosis of pleural effusions. PMID- 7483975 TI - Inferior vena cava thrombosis following percutaneous filter insertion: an unusual cause of haemodynamic compromise. AB - Inferior vena cava thrombosis is a well-recognized complication of filter insertion. We report on two patients who presented with progressive exertion dyspnoea and lipothymia revealing a complete inferior vena cava thrombosis below the filter. Haemodynamic exploration demonstrated that cardiac output could not adapt during upright exercise owing to inappropriate cardiac preload. Since these general manifestations can precede any symptom of lower limb venous stasis, thrombosis of inferior vena cava has to be carefully searched in this setting. The pathogenesis of this unusual complication is reviewed in the light of experimental models of vena cava ligation. PMID- 7483976 TI - Acquired haemophilia in the elderly. AB - The occurrence of an acquired inhibitor of F VIII coagulant activity is a rare cause of a sometimes important bleeding diathesis. Antibodies against F VIII:C can spontaneously occur, mostly in elderly patients. Four elderly patients with such a typical clinical and biological syndrome are reported. No identifiable underlying disease was found and they responded favourably to an immunosuppressive treatment. In one patient plasmapheresis was successful. PMID- 7483977 TI - [2 cases of isolated tricuspid endocarditis following colonic intervention]. AB - We report two cases of tricuspid valve endocarditis with related organisms (Enterococcus faecalis and Gemella morbillorum), some months after colonic operation. This association, without preexistent valvular pathology and without any other classical aetiology of tricuspid endocarditis, has only exceptionally been described. PMID- 7483979 TI - Cytopathology of the respiratory tract. PMID- 7483978 TI - Administration of two vaccines, DTP and Hib, in one single injection. PMID- 7483980 TI - Prognostic factors in bronchoalveolar lavage in 77 patients with bone marrow transplants. AB - OBJECTIVE: To study the prognostic implications of the following factors in bronchoalveolar lavage (BAL) specimens in bone marrow transplant (BMT) patients: number of lymphocytes per high-power field (HPF), presence of hemosiderin-laden macrophages (HLMs), presence of infections and presence of reactive epithelial cellular changes in bronchial and alveolar cells. STUDY DESIGN: A retrospective study of 77 bone marrow transplant patients who required BAL. Each specimen was studied for the number of lymphocytes per HPF, presence of HLMs, presence of infection and presence of reactive epithelial cells. RESULTS: Patients who had < or = 5 lymphocytes per HPF had a much higher death rate than did patients who had > 5 lymphocytes per HPF (P < .001). Eighty-nine percent of patients who had < or = 5 lymphocytes per HPF died within 30 days of undergoing BAL (P < .001). Patients without HLMs had a better prognosis than did patients with HLMs (P < .005). Thirty-six documented viral or fungal infections occurred in 33 patients. Patients with infections had a greater chance of dying than did those who were infection free (P < .05). Multiple logistic regression analysis showed that the previous three factors are statistically significant. Reactive epithelial cellular changes were seen primarily in patients with autologous BMT (11 of 14 cases) and in those with hematologic malignancies, but they had no statistical significance prognostically. CONCLUSION: The following factors in BAL have a good prognostic value: number of lymphocytes > 5 per HPF, absence of HLMs and absence of infections. Reactive epithelial cellular changes have no prognostic value. PMID- 7483981 TI - Diagnosis of Pneumocystis carinii pneumonia by bronchoalveolar lavage in AIDS patients. Comparison of Diff-Quik, fungifluor stain, direct immunofluorescence test and polymerase chain reaction. AB - OBJECTIVE: To assess the sensitivity, specificity and accuracy of Diff-Quik, fungifluor stain, the direct immunofluorescence test (DIFT) and the polymerase chain reaction (PCR) in the diagnosis of Pneumocystis carinii pneumonia (PCP) in human immunodeficiency virus (HIV)-infected patients. STUDY DESIGN: From December 1992 through November 1993, 112 bronchoalveolar lavage fluid (BALF) samples were obtained from 80 HIV-infected patients. BALF samples were processed for cytologic and microbiologic analysis and for PCR. Cytologic examination was carried out on Diff-Quik-stained cytocentrifuge preparations and with May-Grunwald-Giemsa staining and fungifluor staining. For diagnosis of PC infection, DIFT and PCR were used. RESULTS: Thirty-two of 112 acute episodes were caused by P carinii. Diff-Quik had the highest sensitivity (84.8%) as compared to fungifluor stain (60.0%), DIFT (59.4%) and PCR (65.6%). The specificity was 98.7% with Diff-Quik, 100% with fungifluor stain, and 98.6% and 97.3% with DIFT and PCR, respectively. Accuracy was high with every method (94.4% with Diff-Quik, 88.3% with fungifluor stain, 86.7% with DIFT and 87.6% with PCR). CONCLUSION: Diff-Quik is a good diagnostic tool in the diagnosis of PCP. The combination of Diff-Quik and fungifluor stain is recommended because of its cost-effectiveness and because of its rapid diagnosis of severe PCP. PCR and DIFT should be used only on patients judged clinically to have PCP with discrepant results in Diff-Quik and fungifluor stain in BALF samples. PMID- 7483982 TI - Fine needle aspiration cytology of pulmonary Wegener's granulomatosis with biopsy correlation. A report of three cases. AB - BACKGROUND: Pulmonary Wegener's granulomatosis frequently presents clinically as ill-defined nodules on chest radiographs or computed tomographic scans. With refinement of fine needle aspiration (FNA) cytology as a diagnostic tool, pathologists are sometimes asked to render this diagnosis on cytologic material rather than on the more invasive open lung biopsy. Experience is currently limited to a few case reports in the cytopathology literature. We add to this by reporting the findings from five FNA biopsies performed on three patients, with open lung biopsy correlation. CASES: The cytologic findings included a background of necrotic debris, acute and chronic inflammatory cells, multinucleate giant cells and, in one case (case 2), numerous clusters of atypical epithelioid cells mimicking carcinoma. Correlation of the respective open lung biopsy specimens with the FNA material was instructive in revealing the nature of the findings on cytologic preparations. Two cases showed classic histopathologic findings. Case 3 showed a less common presentation, pulmonary hemorrhage, which was reflected in the FNA specimen. CONCLUSION: FNA biopsy cytology is a useful alternative to open lung biopsy in the diagnosis of pulmonary Wegener's granulomatosis but requires careful correlation with the clinical and laboratory findings. PMID- 7483983 TI - Bronchoalveolar lavage for diagnosis of fungal disease. Five years' experience in a southern United States rural area with many blastomycosis cases. AB - OBJECTIVE: To determine the relative frequency of Blastomyces dermatitidis among other microorganisms in bronchoalveolar lavage (BAL) specimens. STUDY DESIGN: The study group consisted of 208 BAL specimens received from 192 patients from March 1988 to August 1993. RESULTS: Forty-seven specimens from 42 patients were positive for pathogenic microorganisms, and 2 other specimens were diagnostic of malignancy. Pneumocystis carinii (23 specimens) was the most common microorganism found in the specimens. Candida spp (10 specimens) was the second most common microorganism, and B dermatitidis (5 specimens) was the third. Cryptococcus neoformans (3 patients), Histoplasma capsulatum (2 patients) and Conidiobolus coronatus (1 patient) were the other fungi detected in BAL. Acid-fast bacilli, cytomegalovirus and herpes simplex virus were also found (1 patient each). Several patients had more than one organism. CONCLUSION: B dermatitidis was the third most common microorganism found in BAL specimens at our hospital. PMID- 7483984 TI - Pulmonary microsporidiosis in a patient with the acquired immunodeficiency syndrome. A case report. AB - BACKGROUND: Microsporidian protozoa are increasingly recognized as pathogenic organisms related to host immunosuppression. CASE: We report a case of pulmonary microsporidiosis detected in aspirated bronchial material and in bronchoalveolar lavage fluid collected from a 26-year-old, male acquired immunodeficiency syndrome patient presenting pulmonary symptoms and known to have intestinal microsporidiosis. The diagnosis was made on cytologic examination of the material processed according to the Papanicolaou technique, evaluated by Giemsa, Gram's, Grocott and chromotrope-2R modified trichrome preparations and by fungi-fluor fluorescent mycology stain. CONCLUSION: Our findings reveal an important fluorescent property of intracellular microsporidian spores, heretofore undescribed in the literature. PMID- 7483985 TI - Bronchoalveolar lavage cytology in pulmonary sarcoidosis. AB - OBJECTIVE: To study the cytomorphologic findings in bronchoalveolar lavage (BAL) fluid in pulmonary sarcoidosis, relationship of the cytologic findings to radiologic stage and cytohistomorphologic correlation. STUDY DESIGN: We retrospectively studied the cytologic findings in BAL specimens from 26 patients who had clinical features and histologic findings consistent with pulmonary sarcoidosis. RESULTS: Clean background; scattered, multinucleate giant cells; epithelioid cells; reactive alveolar macrophages; and a significant population of lymphocytes were observed in 88.5% (23/26) of cases of pulmonary sarcoidosis. A frequent finding was reactive nuclear changes in most multinucleate giant cells and epithelioid histiocytes, consisting of nuclear enlargement, open chromatin and one to two small, acidophilic nucleoli. CONCLUSION: The findings suggest that a BAL specimen in the majority of patients with pulmonary sarcoidosis yields this unique combination of cytologic features. The technique is accurate and economical and may be valuable in the management of such cases. PMID- 7483986 TI - Comparison of histologic stains in the diagnosis of Pneumocystis carinii. AB - OBJECTIVE: To evaluate commonly used histologic stains used to detect Pneumocystis, including Diff-Quik (DQ), Grocott's Gomori methenamine silver (GMS), Papanicolaou, and hematoxylin and eosin (H&E). STUDY DESIGN: The study compared the ability of these stains to detect alveolar casts. RESULTS: Although there was a slightly higher average number of casts detected with DQ, the difference between this stain and GMS was not significant. Both Papanicolaou stain and H&E showed significantly lower averages. Some differences in H&E may be related to differences in preparation. CONCLUSION: Similar results were obtained with DQ and GMS, but DQ offered the advantage of ease of preparation, rapidity and cost-effectiveness. PMID- 7483987 TI - Safe separation of sputum cells from mucoid glycoprotein. AB - OBJECTIVE: To compare the safety and efficacy of homogenizing sputum with deoxyribonuclease I (DNAse I), dithiothreitol (DTT), N-acetyl-L-cysteine, sodium EDTA and trypsin against standard mechanical blending to provide mucus-free, single-cell suspensions for quantitative analysis. STUDY DESIGN: Clinical sputum specimens or cultured human bronchogenic carcinoma cells were preserved in 2% polyethylene glycol/50% ethanol, divided into aliquots, counted and stained (Papanicolaou and avidin-biotin complex immunostained) at baseline. Cells of each aliquot were separated from mucus by the standard physical blending method or by chemical or enzymatic mucus liquefaction. After staining, washing and resuspending in the original volume of polyethylene glycol/ethanol mixture, aliquots were again counted and stained. RESULTS: Cell counts, Papanicolaou staining and immunostaining showed that homogenization of induced, preserved sputum with 0.5 mM DTT is safe and provides mucus-free monolayers for immunocytochemistry and single-cell suspensions for flow cytometry. Mucolysis with 0.5 mM DTT resulted in a significant (16%) increase in cells available. In contrast, mechanical blending resulted in up to a 24% reduction in specimen cellularity. CONCLUSION: Homogenization with low-concentration DTT will probably facilitate the exploration of sputum for protein and gene markers of carcino genesis. PMID- 7483988 TI - Sensitivity of sputum cytology after homogenization with dithiothreitol in lung cancer detection. Two years of experience. AB - OBJECTIVE: To compare the sensitivity of sputum cytology using the dithiothreitol homogenization method (DTT) with that of the standard pick-and-smear (PS) method in lung cancer detection. STUDY DESIGN: In a two-year period, 2,428 sputum specimens were examined. Standard PS smears were prepared as controls; the remainder of the specimens were homogenized with DTT before smears were prepared. The sensitivity of the two methods in detecting positive cases was compared. RESULTS: Among 260 positive cases, the DTT method detected 256, while the PS method detected only 212, representing a difference of 17%. The DTT method also detected 28 more suspicious cases and permitted definitive typing of the tumor in a further 11 cases. CONCLUSION: Homogenization of sputum with DTT is not only a simple and versatile method but also a more sensitive method as compared to the PS method in lung cancer detection. PMID- 7483990 TI - Bronchoscopic diagnosis of malignancy in the lower airway. A cytologic review. AB - OBJECTIVES: To determine the significance of a "suspicious" cytology report, to evaluate diagnostic accuracy for these specimens in the laboratory and to review the correlation of tumor subclassification in cytology versus histology. STUDY DESIGN: Retrospective review and comparison of 269 bronchoscopic cytology and histology specimens from a six-year period, 1984-1990. The relative and combined sensitivities of the sampling methods were studied. RESULTS: There were 17 suspicious cytologic diagnoses during this period. One of 17 was a false suspicious diagnosis, representing 0.03% of all reports on submitted samples. There were no false positive cytologic diagnoses. The majority (94%) of patients with a suspicious cytologic report had a final diagnosis of malignancy. Carcinoma was diagnosed in 244 of the 269 patients. CONCLUSION: Combined bronchoscopic cytology and biopsy was more sensitive (92%) than biopsy alone (82%) in making the initial diagnosis. PMID- 7483991 TI - Cytopathology of pulmonary carcinoid tumors in sputum and bronchial brushings. AB - OBJECTIVE: To study the cytologic manifestations of bronchial carcinoid tumor (BCT) in sputum and bronchial brushing (BB). STUDY DESIGN: A diagnosis of BCT was made by sputum examination in 1 case and by BB cytology in 12 cases with central and typical BCTs. The tumor was suspected in three patients whose sputa were too scanty in diagnostic cells. Three patients with central and typical BCTs yielded only a few tumor cells in BB samples and were suspected of having BCT. In three patients with central and atypical BCT, a diagnosis of oat cell carcinoma was made in two cases and typical BCT in the third case by BB. One case of peripheral and typical BCT was correctly identified by BB. In BB samples the typical BCTs showed tumor cells singly and in loose clusters. They displayed oval nuclei, fine chromatin clumping, micronucleoli and ill-defined, variable cytoplasm. The number of tumor cells with plasmacytoid configuration varied from case to case and was abundant in one patient. The peripheral BCT was predominantly composed of spindle cells and in BB samples showed single and loosely clustered cells with ill defined cytoplasm and cigar-shaped nuclei. Two atypical BCTs yielded pleomorphic cells displaying scant cytoplasm, nuclear molding, fine chromatin and rare micronucleoli, similar to those of oat cell carcinoma, but the smear background was free of necrotic debris. CONCLUSION: BCTs rarely exfoliate their cells in sputum, and bronchial brushing is efficient in detecting BCT cells that show distinctive cytologic features of the tumors. PMID- 7483989 TI - Immunogold-silver staining of cells recovered by bronchoalveolar lavage. A report of three cases demonstrating its value in the diagnosis of pulmonary involvement in hematologic malignancies. AB - BACKGROUND: The diagnosis of pulmonary involvement in hematologic malignancies (PHM) has traditionally necessitated large specimens (usually invasive or surgically obtained biopsies). The development of reliable immunocytologic identification techniques allows diagnosis on noninvasively obtained cellular material, including bronchoalveolar lavage (BAL) fluid. Since 1985, 12 such cases have been reported. CASES: Three cases of B-cell-related PHM were diagnosed by means of BAL fluid cellular analysis using the immunogold-silver staining technique. CONCLUSION: This is the first report showing immunogold-silver staining to be a reliable immunocytochemical identification technique for monoclonal B-cell populations recovered by BAL. PMID- 7483992 TI - Expression of keratins and carcinoembryonic antigen in bronchial squamous metaplasia and lung carcinomas. AB - OBJECTIVE: To determine the incidence of carcino-embryonic antigen (CEA), cytokeratin8.60 (CK8.60) and CK18 expression in sputum smears from patients with various types of lung cancer and in heavy smokers with various degrees of squamous metaplasia without lung cancer. STUDY DESIGN: Fifty-six patients with lung cancer and 149 heavy smokers were investigated. RESULTS: For all markers a progressive increase in the numbers of positives was observed as the degree of squamous metaplasia increased in both groups. CK8.60 was positive in 95% of heavy smokers (benign histology) with atypical squamous metaplasia but in only 67% of malignant cells. Combinations of positive CEA and CK8.60 were frequent, while CK18 positivity tended to be solitary. CONCLUSION: Positive staining might be indicative of an increased risk of malignant transformation regardless of cytology, but positivity of markers is not of use in the diagnosis of malignant histology. PMID- 7483993 TI - Fine needle aspiration biopsy of pulmonary hamartomas. Radiologic, cytologic and immunocytochemical study of 15 cases. AB - OBJECTIVE: To review the radiographic, cytologic and immunocytochemical features of pulmonary hamartomas (PHs) diagnosed on fine needle aspiration biopsy (FNAB). STUDY DESIGN: Fifteen consecutive cases of PH, diagnosed on FNAB between January 1987 and February 1993 and confirmed by surgery or follow-up, were studied. In two additional cases PH was offered as a differential diagnosis and was excluded on follow-up. Clinical notes, radiographs, cytologic smears and cell block sections stained routinely and with antibody to S-100 protein, as well as histologic slides, were reviewed. RESULTS: All cases of PH showed common radiographic features, including peripheral location, round or oval shape, sharp edges and size < 3 cm. Cytologic diagnosis of PH was based on recognition of mesenchymal component, characterized by fibromyxoid stroma, present in 94% of FNABs. Chondroid material was present in 75% of aspirates. In all cases of proven PH, fibromyxoid material showed S-100 protein positivity, characterized by finely granular, brown staining of the stellate cells. In two cases proven not to be hamartomas, the material, suspected to be fibromyxoid or chondroid, failed to show S-100 protein positivity. CONCLUSION: These findings confirm the value of FNAB in the diagnosis of PH. Immunocytochemical staining with antibody to S-100 protein is a useful diagnostic tool in confirming the cartilaginous nature of PH. The cytologic findings should be correlated with radiologic and clinical findings before a definitive diagnosis of PH is rendered on FNAB material. PMID- 7483994 TI - Mixed small cell/large cell carcinoma of the lung. Report of a case with cytologic features and ultrastructural correlation. AB - BACKGROUND: Mixed small cell/large cell carcinoma of the lung, a chemotherapy- and radiation-resistant subtype of small cell lung cancer, constitutes 4-6% of small cell lung cancer but has been described rarely in the cytopathology literature. CASE: A 64-year-old man presented with a left hilar mass. A concurrent transbronchial biopsy and postbiopsy bronchial washing were performed. The latter presented as loosely cohesive cells, scattered singly, in small clusters, in monolayer sheets and in a perivascular arrangement. The tumor exhibited a wide spectrum of cytomorphology: small cells with pyknotic nuclei and scanty cytoplasm were admixed with larger cells with vesicular nuclei, prominent nucleoli and abundant cytoplasm. Nuclear shape included oval, spindle and peg. Nuclear size was also highly variable. The tumor was retrieved from the transbronchial biopsy and processed for ultrastructural study. CONCLUSION: Based on cytologic-ultrastructural correlations. the seemingly "mixed" morphology may have resulted from the rapid degeneration of a single clone of tumor cells: the viable tumor cells were the "large" cells, and the dying cells were the "small" cells. A novel ultrastructural observation is that the perivascular tumor cells developed peculiar, fingerlike cytoplasmic processes abutting an undulating basement membrane along the thin-walled blood vessel. PMID- 7483995 TI - Bronchioloalveolar carcinoma in progressive systemic sclerosis. Report of a case diagnosed by fine needle aspiration cytology. AB - BACKGROUND: Progressive systemic sclerosis (PSS) is a systemic disorder involving many internal organs, including the lungs. The occurrence of a pulmonary tumor has occasionally been reported, especially in long-standing cases. Bronchioloalveolar carcinoma is the most prevalent histologic type. CASE: A 35 year-old male with a 12-year history of PSS presented with exertional dyspnea and pain in the left side of the chest. Fine needle aspiration cytology (FNAC) showed abundant cellular material with large, monolayered sheets and papillary structures. The neoplastic cells had uniform round to oval nuclei and spacious cytoplasm. Some had intranuclear vacuoles. Phosphotungstic acid-hematoxylin (PTAH) stain revealed structures in the form of prominent cell membranes rimming the free surfaces of the tumor cells. The overall cytologic picture was interpreted as consistent with bronchioloalveolar carcinoma. The patient's condition did not permit surgery. CONCLUSION: In this case the development of bronchioloalveolar carcinoma occurred over a 12-year period following the diagnosis of PSS. This case emphasizes diagnosis of the tumor solely by FNAC. PTAH stain was also helpful in the cytologic diagnosis. PMID- 7483996 TI - Echinococcus hooklets in sputum: sinus tract extension of a liver hydatid cyst to lung. PMID- 7483997 TI - A clinical trial of the AutoPap 300 QC system for quality control of cervicovaginal cytology in the clinical laboratory. AB - OBJECTIVE: False-negative reports of cervicovaginal (CV) smears (FNs) due to interpretive errors pose a significant and persistent problem in cytology. Using routinely prepared CV smears, the AutoPap 300 QC System, an automated cytology screening device, can provide an enriched population of slides for manual rescreening. In this clinical trial, the performance of the AutoPap System was assessed and compared to current random quality control (QC) selection techniques. STUDY DESIGN: A total of 3,487 "within normal limits" cases were rescreened, both manually and by the AutoPap 300 QC System. Detected FNs were classified by three different methods: cytotechnologist rescreen, internal discrepancy panel and external discrepancy panel. The sensitivity of the AutoPap System to these FNs was established at five QC score thresholds. RESULTS: By cytotechnologist rescreening, 106 FNs were identified. Subsequent internal and external discrepancy panel reviews confirmed the FNs in 80 cases and 86 cases, respectively. At a 10% review rate, the AutoPap System identified approximately 50% of the squamous intraepithelial lesion FNs. CONCLUSION: The AutoPap 300 QC System is a sensitive automated cytology system that has the potential of yielding five times as many FNs as a 10% random rescreen. PMID- 7483998 TI - Endocervical columnar cell intraepithelial neoplasia. I. Discriminating cytomorphologic criteria. AB - OBJECTIVE: To identify specific cytomorphologic characteristics of mild, moderate and severe atypia of endocervical columnar cell lesions. STUDY DESIGN: The study group consisted of 55 smears from 53 women known to have columnar cell atypias of variable severity. The smears were reviewed by five observers in terms of 46 architectural, cellular and nuclear features. Review diagnoses were given together with the scoring for a large number of cellular characteristics. RESULTS: Nuclear chromatin distribution, variation in cellular and nuclear size, cytoplasmic eosinophilia and such architectural features as cell crowding, cluster formation, formation of gland-like structures and pseudostratification appeared to be of primary diagnostic importance for discriminating between no abnormalities, different grades of severity of endocervical intraepithelial neoplasia and adenocarcinoma. With acceptance of one grade of difference in grades of severity, a correct diagnosis was made in 87.6% of cases. CONCLUSION: With the application of specific architectural, cellular and nuclear features, the cytologic detection of different grades of endocervical intraepithelial neoplasia and of adenocarcinoma proved to be dependable and reproducible, with accuracy that at least equaled that of detecting squamous (metaplastic) lesions. PMID- 7483999 TI - Endocervical columnar cell intraepithelial neoplasia. II. Grades of expression of cytomorphologic criteria. AB - OBJECTIVE: To determine the relationship between the grade of expression of discriminating cytomorphologic architectural, cellular and nuclear features and the degree of endocervical columnar cell atypia. STUDY DESIGN: The study group consisted of 55 smears from 53 women known to have endocervical columnar cell atypias of variable severity. The smears were reviewed by five observers by means of 46 architectural, cellular and nuclear features. Seventeen of the 46 evaluated features were scored as revealing three grades of expression: slight, moderate and marked. RESULTS: Grades of expression of cellular and nuclear characteristics used in the evaluation of endocervical columnar cell abnormalities were identical to the characteristics used in the diagnosis of squamous (metaplastic) lesions. As such the features were extremely useful, with the exception of hyperchromasia, which was less prominently expressed. Furthermore, the results on architectural features revealed that the expression grades slight and moderate were indicative of mild and moderate endocervical columnar cell atypia, whereas the marked expression grade was strongly related to the severe intraepithelial lesions and adenocarcinoma. CONCLUSION: Characteristic cytomorphologic features at different grades of expression proved to be reliable light microscopic parameters in the diagnosis of different grades of endocervical columnar cell abnormalities. The accuracy of diagnosis at least equaled that of detecting squamous (metaplastic) lesions. PMID- 7484000 TI - A one-year audit of fine needle aspiration cytology of breast lesions. Factors affecting adequacy and a review of delayed carcinoma diagnoses. AB - OBJECTIVE: To compare the rate of inadequate specimens in a triple-diagnosis breast clinic with that of specimens taken by clinicians. STUDY DESIGN: Breast aspirates obtained in 1993, 2,923 specimens, were reviewed. Of these, 1,515 (51.9%) were taken in a specialist clinic with a triple approach (clinical examination, imaging and aspiration cytology). Palpable lesions were aspirated by a cytopathologist and nonpalpable lesions by a cytopathologist and radiologist in cooperation using imaging techniques. The rate of inadequate specimens was determined and compared with the rate of the remaining specimens, which were sent to the laboratory by clinicians. RESULTS: In the breast clinic, the inadequacy rate was 3.8% (n = 58). The inadequacy rate for the 1,408 mailed breast aspirates was 14.8% (n = 209). A cancer diagnosis was recorded in 13.5% (n = 395). Six carcinomas were diagnosed late because of inadequate specimens. CONCLUSION: The inadequacy rate was lower in cytology samples taken in a specialized breast clinic than in samples mailed to the laboratory by clinicians. In order to increase the detection rate of malignancies, clinicians performing fine needle aspiration should be trained in the aspiration technique and smear preparation. PMID- 7484001 TI - Mesenchymal chondrosarcoma of the retroperitoneum. Report of a case diagnosed by fine needle aspiration biopsy with immunohistochemical, electron microscopic demonstration of S-100 protein in undifferentiated cells. AB - BACKGROUND: Mesenchymal chondrosarcoma (MC), first described in 1959 by Lichtenstein and Bernstein, is an uncommon tumor of the soft tissues and bone defined by the coexistence of nests of well-defined cartilaginous tissue within a proliferation of primitive mesenchymal cells. CASE: A 27-year-old, white female consulted a physician for a left lumbar mass associated with sciatica-type pain, asthenia, anorexia and weight loss. Abdominal computed tomography showed a 20-cm, retroperitoneal neoplasm that occupied the lower half of the left abdominal flank. Fine needle aspiration biopsy showed clusters of small round cells mixed with islets of mature cartilage. Immunomarking for electron microscopy, carried out with 15 nm of A protein/colloidal gold, showed specific immunoreactivity to S 100 protein in the nuclear chromatin, nucleoli, Golgi apparatus, plasma membrane and cytosol of cells with more developed organelles. CONCLUSION: The diagnosis of MC rests on recognition of the biphasic component of small, undifferentiated cells and cartilage islets. In aspirates with only undifferentiated cells, demonstration of ultrastructure-specific immunoreactivity to S-100 protein may contribute to the final diagnosis. PMID- 7484002 TI - Asteroides hyalosis. A case report. AB - BACKGROUND: Asteroides hyalosis is a degenerative condition of the vitreous of unknown etiology in which calcium soaps accumulate on vitreous fibrils in the eye. Cytologically, the condition needs to be differentiated from foreign body reactions, synchysis scintillans and proteinaceous coagula. CASE: A 69-year-old male presented with a history of diminished vision and floaters in the right eye. Examination of the vitreous fluid revealed roughly spherical structures. When examined with polarized light, these structures showed central, birefringent, crystalline particles. CONCLUSION: The cytologic appearance of asteroides hyalosis is characterized by round asteroid bodies ranging in size from 30 to 80 microns in diameter. When polarized, these structures show central, birefringent, crystalline particles. Special stains may be helpful in the diagnosis. PMID- 7484004 TI - Cytologic grading of breast carcinoma. PMID- 7484003 TI - Cytology of metastatic leiomyosarcoma. A case report. AB - BACKGROUND: Metastasis of leiomyosarcoma is not uncommon. Determination of the primary site may be confused with the presenting symptoms and even the known pathology. CASE: A 62-year-old man presented with chest pain, a productive cough and a lung mass on radiography. Fine needle aspiration of the tumor showed pleomorphic tumor cells. The patient began to have abdominal symptoms one year after presentation. Jejunal leiomyosarcoma was proven after an emergency exploratory laparotomy for mechanical ileus. Sputum cytology disclosed tumor cells one month before death. Necropsy of the lung mass demonstrated findings similar to those of the intestinal tumor microscopically. CONCLUSION: Without laparotomy, our patient would have been diagnosed as having a primary pulmonary leiomyosarcoma by the presenting symptoms and known pathology. PMID- 7484006 TI - Maltese crosses in Cryptococcus neoformans. PMID- 7484007 TI - Diagnosis of Langerhans cell histiocytosis by fine needle aspiration cytology. PMID- 7484005 TI - Hyaluronan in the background of a fine needle aspiration biopsy specimen from the breast: a possible diagnostic clue. PMID- 7484008 TI - Retroperitoneal extragonadal germ cell tumor diagnosed by needle aspiration cytology and electron microscopy. PMID- 7484009 TI - New variant of congenital dyserythropoietic anemia with trilineage myelodysplasia. AB - We report the case of a male infant with a variant of congenital dyserythropoietic anemia (CDA), who developed severe hyperbilirubinemia on the day of birth, subsequent severe anemia, and hyperferritinemia. Bone marrow and laboratory examinations revealed features of CDA including trilineage myelodysplasia and erythroblasts with a binucleated nuclear morphology and ineffective erythropoiesis. The CDA in this patient was assumed to be a new variant type because of: the lack of internuclear chromatin bridges in the erythroblasts with abnormal nuclear morphology; a negative acid serum test; the presence of erythrocyte antigen I, and the effect of splenectomy. Trilineage myelodysplasia in CDA is not known. An abnormality in the stem cells was suggested to be the cause of CDA in this case. PMID- 7484010 TI - Hairy cell leukemia variant. AB - A 59-year-old man presented with lymphocytosis with huge splenomegaly. The abnormal lymphocytes had a high nucleoplasm:cytoplasm ratio, a prominent nucleolus and hairy cytoplasmic projections. Immunophenotyping revealed B-cell leukemia with negative reactions to CD5 and CD25. Cytogenetic study showed 46,XY,der(5)t(5;6)(q35;p21), del(7)(p13)/46,idem,add(22)(q13). The patient did not respond to chlorambucil and a combination of cyclophosphamide, vincristine and prednisolone. Splenic irradiation induced partial remission. He developed progressive anemia and thrombocytopenia and died of Escherichia coli septicemia 33 months after the diagnosis of hairy cell leukemia variant. PMID- 7484011 TI - Normal granulocyte function but impaired monocyte function in a patient with aplastic anemia. AB - Granulocytopenia is an invariable finding in aplastic anemia (AA) and bacterial infections are a frequent complication and major cause of death in patients suffering from this bone marrow disorder. Using well-established assays, we determined the chemotaxis of granulocytes and monocytes as well as the phagocytic capacity of monocytes and oxidative metabolism of granulocytes in a patient with aplastic anemia of unknown etiology. Our results indicate a normal granulocyte oxidative metabolism and granulocyte chemotaxis towards formyl-leucyl-methionyl phenylalanine (fMLP), whereas monocyte chemotaxis and phagocytic capacity were markedly reduced in this patient. PMID- 7484012 TI - c-kit-positive cells and CD34-positive cells in cryopreserved peripheral blood stem cell samples. PMID- 7484013 TI - Uric acid, creatinine and urea in normal, glucose-6-phosphate dehydrogenase deficient and Hb SS Saudi subjects. PMID- 7484014 TI - Management of stage IE primary gastric lymphoma. AB - From 1975 to 1991, stage IE gastric lymphoma was diagnosed or treated in 40 patients at our institution. Thirty-five evaluable patients were the subject of this analysis (median age, 63 years). All histologic specimens were reviewed and classified according to the working formulation. Immunohistochemical stains were performed in 33 of 35 cases. The following histologic subtypes were identified: 26 patients had diffuse large cell, 2 had diffuse mixed cell, and 7 had mucosa associated lymphocytic lymphoma (small lymphocytic lymphoma with features of MALT lymphoma). Of the 35 patients, 30 (86%) underwent laparotomy, with subtotal or total gastrectomy performed in 29. After gastrectomy, 17 patients had radiation therapy, 3 had chemotherapy, 2 had radiation plus chemotherapy, and 7 had no further treatment. Five patients were diagnosed with endoscopic biopsy and had radiation therapy, chemotherapy, or both without further surgical treatment. The median follow-up period was 69.2 months. The commonest presenting symptom was pain. The 5-year disease-free survival was 90% for the entire group and 88% for the 28 patients with working formulation intermediate histology. For patients with MALT lymphoma, the 5-year disease-free survival was 100%. The prognosis for patients with stage IE working formulation intermediate-histology gastric lymphoma is good and for those with stage IE MALT lymphoma, excellent. PMID- 7484015 TI - Viridans streptococcal shock syndrome during bone marrow transplantation. AB - Of 320 patients receiving a marrow transplant at the Hospital de Sant Pau between 1986 and 1992, 12% developed viridans streptococcal bacteremia during severe neutropenia. Five of these patients (13%) developed a rapidly progressive fatal shock syndrome characterized by bilateral pulmonary infiltrates, acute respiratory failure (ARDS) and septic shock early in the transplantation course (6 or 7 days posttransplantation). All patients were transplanted for acute leukemia in remission, and 2 received an allogeneic and 3 an autologous transplant. Four of these subjects were younger than 15 years of age and all had received cyclophosphamide and total body irradiation as conditioning regimen for marrow transplantation. All 5 patients died, and postmortem examinations revealed diffuse pulmonary lesions characteristic of the ARDS. These observations contribute to defining the clinical and pathologic characteristics of this serious complication of intensive anticancer treatment. PMID- 7484016 TI - Alcoholic dyshaematopoiesis: morphological features of alcohol-induced bone marrow damage in biopsy sections compared with aspiration smears. AB - We investigated trephine biopsies from 118 alcoholic patients in order to establish whether or not the known alcohol-induced alterations seen in smears are seen in bone marrow tissue sections, and whether new features might emerge, which could be suitable for defining the histological picture of a condition to be known as 'alcoholic dyshaematopoiesis'. The main changes involve erythropoiesis and iron metabolism. They can be attributed in part to destabilization of the red cell membrane. While these alterations are clearly visible in tissue sections, aspiration smears are needed to detect other features, especially ring sideroblasts. PMID- 7484017 TI - Multidrug resistance in B-cell chronic lymphocytic leukemia. AB - Multidrug resistance (MDR) was investigated in peripheral blood cells isolated from 40 patients with B cell chronic lymphocytic leukemia (B-CLL) and from 7 healthy volunteers, using a flow cytometric assay that detects cellular efflux of the fluorescent dye rhodamine 123 (Rh 123), which has been demonstrated to be transported from the cell by the P-glycoprotein pump. The proportion of B leukemic cells effluxing Rh 123 and thus displaying MDR was low (14 +/- 17%) in B CLL and in only 4 cases did the contingent of B leukemic cells showing MDR represent more than 30% of the total leukemic cells. In contrast, a higher proportion of cells effluxing Rh 123 (44 +/- 13%) was demonstrated in normal B lymphocytes. No statistical correlation was found between the number of leukemic B cells displaying MDR and clinical parameters or previous treatment. These results clearly suggest that MDR activity is usually low in B-CLL. PMID- 7484018 TI - Prognostic value of nephromegaly at diagnosis of childhood acute lymphoblastic leukemia. AB - In order to assess the prognostic value of nephromegaly (kidney enlargement > or = 2 SD from mean for age) in children at diagnosis of acute lymphoblastic leukemia (ALL), kidney size was investigated by intravenous pyelogram (IP), according to the method of Ekl of and Ringertz, in 101 children diagnosed with ALL in the period of 1975-1983. In the same period 103 additional patients with ALL were not investigated with IP for logistical reasons. Characteristics at the diagnosis of ALL in the patients of the investigated and non-investigated (with IP) groups were quite similar and event-free-survival (EFS) curves superimposable. Eighty-nine (50 males and 39 females, median age 61 months) out of 101 patients underwent IP before starting any therapy and were evaluated in the study; 27 (30.3%) presented with nephromegaly (bilateral in 13 cases and unilateral in 14). With a median follow-up time of 14.4 years (range 11-19 years), the EFS at 15 years from diagnosis was 18.5% (SE 9.5) in the nephromegalic group and 48.4% (SE 6.4) in the non-nephromegalic group. The association of poorer EFS with nephromegaly was confirmed when the comparison was adjusted by white blood cell count and age at diagnosis in a Cox regression model. Nephromegaly at diagnosis of childhood ALL may have an independent prognostic value in patients treated with 'not intensive' protocols; however, this finding should be confirmed in patients treated uniformly with contemporary intensive protocols. PMID- 7484019 TI - A case of Crow-Fukase syndrome with elevated soluble interleukin-6 receptor in cerebrospinal fluid. Response to double-filtration plasmapheresis and corticosteroids. AB - We report the case of a 33-year-old Japanese male who presented with thrombocytosis, lower limb edema, severe polyneuropathy with elevated cerebrospinal fluid (CSF) protein level and serum IgA lambda monoclonal component, fulfilling the manifestations of Crow-Fukase syndrome. A high level of soluble interleukin-6 receptor in the CSF was also found, which fluctuated in parallel with the clinical course. Initial treatment with double-filtration plasmapheresis (DFPP) reduced the serum IgA paraprotein level with improvement of the sensory component of the polyneuropathy and decrease of soluble interleukin-6 receptor in the CSF. The remaining clinical features waned off after steroid treatment. The possible role of interleukin-6 in the pathogenesis of the Crow Fukase syndrome and the utility of DFPP treatment are discussed. PMID- 7484021 TI - Sclerosing IgA multiple myeloma. AB - A case of IgA multiple myeloma associated with myelofibrosis and radiological evidence of diffuse osteosclerosis from the disease onset is reported. Bone marrow trephine biopsies performed before and after chemotherapy treatment for myeloma showed grade 4 collagen fibrosis of the bone marrow, thickened bony trabeculae and the presence of plasma cells, both mature and immature. Serum electrophoresis revealed an IgA lambda-paraprotein. Throughout the course of the disease, there was persistent radiological evidence of osteosclerosis, although several lytic lesions appeared late in the disease process. The patient died 5 years after presentation, during an episode of septicaemic shock. It is speculated that cytokine(s) released by the neoplastic plasma cells may stimulate a fibroblastic reaction within the marrow, which subsequently undergoes bony metaplasia resulting in osteosclerosis. PMID- 7484020 TI - Hemolytic anemia in a patient with subacute bacterial endocarditis due to Streptococcus sanguis. AB - We describe a case of hemolytic anemia with subacute bacterial endocarditis due to Streptococcus sanguis. The major hematological features of the patient were a normocytic anemia with reticulocytosis, an elevation of serum lactate dehydrogenase, and a marked reduction of serum haptoglobin. All these signs of hemolytic anemia fluctuated in parallel with changes in inflammatory activity and were dramatically improved by the administration of benzylpenicillin, suggesting that the infection with S. sanguis itself was the cause of hemolytic anemia in this case. PMID- 7484022 TI - Adenosine increases the cutaneous heat pain threshold in healthy volunteers. AB - Adenosine is an endogenously produced substance which in animal experiments exerts anti-nociceptive effects. In humans, algesic effects have been presented following exogenous adenosine administration. A recent study on anaesthetized patients, however, suggested an anti-nociceptive effect during i.v. adenosine. We have studied the pain-reducing effect in healthy volunteers using adenosine 50-80 micrograms.kg-1.min-1 (n = 10), morphine 0.1 mg.kg-1 (n = 5), adenosine 50 micrograms.kg-1.min-1 + morphine 0.1 mg.kg-1 (n = 6), and ketamine 0.1 mg.kg-1 (n = 5); all drugs given i.v., single-blind. Quantitative sensory tests (QST) revealed a significantly increased cutaneous heat pain threshold following adenosine. No effect was seen following ketamine or morphine. Suprathreshold heat pain perception was unchanged in all subjects. Furthermore, warm and cold perception thresholds were not influenced significantly by any drug. Adenosine, morphine and ketamine produced well-known side-effects but of a mild intensity not necessitating any treatment. The present results show that i.v. adenosine can provide a modest but selective increase of cutaneous heat pain thresholds, suggesting a pain-reducing capacity of adenosine in humans. PMID- 7484023 TI - Desflurane: a new volatile anesthetic for cesarean section. Maternal and neonatal effects. AB - Desflurane, a new volatile anesthetic agent with low blood/gas solubility, has recently been studied in clinical and animal trials but its use in obstetrics has not been adequately evaluated. This prospective study was undertaken to evaluate the maternal and neonatal effects of desflurane in obstetrical patients. Seventy five healthy parturients undergoing primary or repeat cesarean section were randomly assigned to one of three groups of 25 each, end-tidal 3% desflurane, 6% desflurane or 0.6% enflurane, combined with 50% N2O and O2. All patients had rapid sequence induction of anesthesia with thiopentone sodium followed by succinylcholine for tracheal intubation. After delivery, anesthesia was maintained with reduced concentration of desflurane or enflurane with 67% N2O in O2, supplemented by butorphanol tartrate. Maternal hemodynamic parameters, blood loss and maternal awareness during surgery were monitored. Neonatal outcome was evaluated by Apgar scores, neurological and adaptive capacity scores (NACS), cord blood gas and acid-base status, and time to sustained respiration (TSR). Maternal blood loss did not differ significantly between the three groups and none of the patients developed intraoperative awareness. All three groups responded to psychomotor performance equally fast. Patients in all three groups developed transient hypertension and tachycardia during induction of anesthesia which returned to baseline values in approximately 5 min. Neonatal outcome was equally good in the three groups. More neonates in the 6% desflurane group had TSR > 90 s compared to the 3% desflurane group (P < 0.05).(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7484025 TI - Synergism between mivacurium and pancuronium in adults. AB - Mivacurium could be a useful agent as a final dose of a muscle relaxant following pancuronium if only additivity exists between these agents. We examined the interaction between mivacurium and pancuronium in 70 patients (ASA I-II) during propofol-alfentanil-N2O-O2 anaesthesia. Neuromuscular function was monitored by adductor pollicis EMG. Firstly we established dose-response curves for mivacurium and pancuronium. Thereafter, 20 patients received a combination of 0.5 times the ED50 doses of mivacurium and pancuronium (cMP) determined in the first part of this study. Patients were randomized to receive the cMP to the same IV-line (n = 10) or to two separate IV-lines in opposite hands (n = 10). ED50 values for mivacurium and pancuronium were 57.7 and 37.1 micrograms kg-1, respectively. Maximal neuromuscular block following the cMP was 91.8 +/- 5.0% (mean +/- SD). This was highly significantly different from the estimated 50% NMB if only additivity exists between mivacurium and pancuronium (P = 0.0001). After the cMP, the 25-75% recovery time was 9.4 +/- 1.3 min and the time to train-of-four ratio of 0.70 was 35.8 +/- 5.4 min. There was no statistical difference in any recorded neuromuscular parameter between the two subgroups receiving mivacurium and pancuronium to the same or to opposite hands (P > 0.40). We conclude that a significant synergism exists between mivacurium and pancuronium which may indicate that mivacurium does not produce a short-acting NMB if given after pancuronium. We do not recommend using mivacurium together with pancuronium. PMID- 7484026 TI - Statistical aspects in studies of preoperative fluid intake and gastric content. AB - Twelve randomised studies of preoperative intake of clear fluids and gastric content were reviewed and presented together with a separate study performed by the authors. A common deficiency in design was the omission of detailed information concerning randomisation procedures, distribution of data, and choice of statistical methods. Only one reviewed study included power calculations and only 3 reported 95% C.I. (Confidence Intervals). Deficiencies in design and data collection rendered the conclusion of two studies invalid. Incorrect use of statistical tests in some of the remaining studies may have resulted in non significant differences becoming statistically significant. This, however, did not change the main conclusion of a meta-analysis that intake of clear fluids up to 2 hours before general anaesthesia was safe as fluid intake either decreased or had no effect on gastric fluid volume. PMID- 7484024 TI - A new method of using gas exchange measurements for the noninvasive determination of cardiac output: clinical experiences in adults following cardiac surgery. AB - New mathematical algorithms have been applied to a computer controlled closed breathing circuit system for non-invasive measurement of cardiac output (COniv). This system has been described in an animal study. Forty patients were studied 5 and 18 hours after cardiac surgery using the thermodilution technique as the reference (COtd). The variables entered into the algorithms for COniv were oxygen uptake, carbon dioxide elimination, end-tidal carbon dioxide partial pressure, tidal volume and arterial oxygen saturation. Mixed venous carbon dioxide partial pressure was obtained from an automatically implemented short rebreathing manoeuvre. Pulmonary perfusion was calculated by a modified Fick equation for carbon dioxide and the shunt flow added to obtain COniv. During mechanical ventilation, there was a good agreement between COtd and COniv (r = 0.8). The bias was -0.14 l/min and the precision was 0.77 l/min. The reproducibility of COniv was 0.03 l/min and for COtd -0.03 l/min with a standard deviation of the difference being 0.35 l/min for COniv and 0.31 l/min for COtd. In awake, but sedated extubated patients, the method proved unsatisfactory on account for uneven tidal volumes and difficulties with leakage around the mouth piece. We conclude that this new technique provides reliable and reproducible measures of cardiac output in sedated, ventilated patients. PMID- 7484027 TI - Comparison of the effects of neostigmine and edrophonium on the duration of action of suxamethonium. AB - Rapid sequence induction of anaesthesia necessitating the use of suxamethonium may occasionally be needed soon after antagonism of neuromuscular block with anticholinesterase agents. The onset and duration of action of 1 mg kg-1 of suxamethonium was recorded in groups of 10 patients each, 5 or 10 min after the administration of edrophonium 1 mg kg-1 or neostigmine 40 micrograms kg-1 given for the antagonism of atracurium-induced neuromuscular block. Plasma cholinesterase activity was measured before, and 5 and 10 min after the administration of the anticholinesterases. A further 10 patients received suxamethonium 1 mg kg-1 without prior atracurium or anticholinesterase administration to serve as controls. The onset of action of suxamethonium was significantly prolonged when administered 5 min after both anticholinesterases, compared to the control group (P < 0.01). Recovery of suxamethonium block was delayed significantly after neostigmine, compared to both the edrophonium and the control groups (P < 0.05-0.001). Plasma cholinesterase activity was significantly reduced with the use of neostigmine but not with edrophonium (P < 0.001). PMID- 7484028 TI - Epidural anesthesia modifies cardiovascular responses to severe hypoxia in dogs. AB - Hypoxia is a critical and sometimes fatal complication of anesthesia. Since there is little information on the cardiovascular response to hypoxia during epidural anesthesia, we assessed the effects of epidural anesthesia on the cardiovascular response to hypoxia and on survival in dogs. We randomly assigned 36 mongrel dogs to one of three groups according to the anesthetic technique used: the thoracic group (n = 12) received thoracic epidural anesthesia plus general anesthesia, the thoracolumbar group (n = 12) received thoracolumbar epidural anesthesia plus general anesthesia, and the control group (n = 12) received general anesthesia alone. We monitored hemodynamics and plasma catecholamine concentrations and assessed survival in these groups during normocapnic hypoxia (FiO2, 0.09 for 120 min). During hypoxic challenge, PaCO2 and PaO2 values were similar in all groups. In both groups that received epidural anesthesia, heart rate, systolic and diastolic arterial pressures, and plasma epinephrine and norepinephrine concentrations were lower and arterial pH was greater than in the control group. There was no significant difference in survival among groups. Epidural anesthesia modified both the physiologic cardiovascular and catecholamine responses to hypoxia. Epidural anesthesia of the thoracic region did not appear to accelerate cardiac arrest, but it attenuated the development of metabolic acidosis during hypoxia. PMID- 7484029 TI - Autotransfusion after open heart surgery: the oxygen delivery capacity of shed mediastinal blood is maintained. AB - Autotransfusion of mediastinal shed blood after open heart surgery has become a common and accepted procedure in reducing the need for homologous transfusion during the last 15 years. The objectives of the present study were to investigate the oxygen delivery capacity of autotransfused shed mediastinal blood, compared to patient-blood, during cardiopulmonary bypass and in the postoperative period. Ten consecutive patients undergoing elective cardiac surgery were studied. Mediastinal shed blood was collected in the cardiotomy reservoir and retransfused during the first 18 postoperative hours. The oxygen delivery capacity of the blood to the tissues was calculated by use of the oxygen status algorithm (OSA 2.0) programme and measurement of the 2,3-diphosphoglycerate (2,3-DPG) concentration. Autotransfusion volume ranged from 450-1530 ml per patient (median 824 ml). Shed blood had a mean haemoglobin level of 8.8 g/dl and 7.4 g/dl at 1 h and 6 h of autotransfusion, respectively. There were no significant changes of 2,3-DPG concentration in the patient-blood during cardiopulmonary bypass or after autotransfusion compared to preoperative values. P50 for oxygen (3.6 and 3.6 kPa) and 2,3-DPG concentrations (5.3 and 5.1 mikromol/ml erythrocyte) in shed mediastinal blood (1h and 6h postoperatively) were not significantly different compared to patient-blood. The results demonstrate that the oxygen delivery capacity of shed mediastinal blood is maintained and that the oxygen affinity of patient-blood is not influenced by autotransfusion. PMID- 7484030 TI - Ketamine inhibits contractile responses of intestinal smooth muscle by decreasing the influx of calcium through the L-type calcium channel. AB - During the inflammatory response, tissues release histamine (H), substance P, serotonin (5-HT), prostaglandins and kinins, agents that mediate manifestations of inflammation such as pain, vasodilation, increased capillary permeability and smooth muscle contraction. In this study we investigated whether racemic (R[+]) ketamine (K) and its isomers are spasmolytic on intestinal smooth muscle contracted by inflammatory mediators, and whether the spasmolytic effect of K is related to changes in calcium influx through the L-type calcium channel or to an interaction of K with opioid receptors. We measured the contractions of guinea pig ileum mounted in an organ bath containing Tyrode's solution gassed with 95% O2/5% CO2 at 37 degrees C. In the first protocol we determined the effect of K and its isomers on contractions induced by five mediators: 10(-7) M H, 10(-8) M substance P, 10(-8) M neurokinin A, 5 x 10(-9) M bradykinin and 5 x 10(-7) M 5 HT. For each of these mediators, we plotted concentration-response curves for the inhibitory effect of K, and from regression fitting of these curves, we calculated the IC50 concentration of K that inhibited the contraction by 50%). In the second protocol we measured the contraction induced by the calcium ionophore A23187 (5.0 x 10(-6) M), both alone and after 1.8-7.2 x 10(-4) M R(+/-)K. Then we examined how the inhibition caused by R(+/-)K was affected by increases in the concentration of extracellular calcium by adding calcium (1.8-7.2 x 10(-3) M).(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7484031 TI - Comparison of vascular effects of ropivacaine and lidocaine on isolated rings of human arteries. AB - Ropivacaine is a new local anaesthetic agent. Previous animal studies have indicated that vasoconstrictor effects are elicited by ropivacaine in vitro and subcutaneously and that it produces blanching of the skin if injected subcutaneously in humans. Lidocaine is a widely used local anaesthetic reported to exert a biphasic effect on the microvasculature with contraction at low concentrations and relaxation at high concentrations. There is a need for pharmacologic tools able to counteract local arterial vasoconstriction. In this study, the contractile effect of ropivacaine and lidocaine were investigated in vitro on isolated human arteries. Experiments were performed on 43 internal mammary artery (IMA) rings obtained from 22 patients and on 14 radial artery (RA) rings from 7 patients. The rings were mounted in organ baths and isometric contractile activity was measured. Experiments were conducted by cumulative adding ropivacaine or lidocaine (1.5 x 10(-5) M; 4.5 x 10(-5) M; 1.5 x 10(-4) M; 4.5 x 10(-4) M; 1.5 x 10(-3) M; 4.5 x 10(-3) M; 1.5 x 10(-2) M) to the organ baths. The endothelium was mechanically removed in 19 IMA rings and in 9 RA rings. Ropivacaine and lidocaine produced a biphasic response with contraction at low concentrations (1.5 x 10(-5)-1.5 x 10(-3) M) and release of the maximal contraction at higher concentrations. No statistically significant differences in contractile or relaxing effects were seen between the two drugs. Removal of the endothelium did not significantly affect contractile activity. In this study of human mammary artery preparations, ropivacaine is not a stronger vasoconstrictor than lidocaine. PMID- 7484032 TI - Paravascular lumbar plexus block: block extension after femoral nerve stimulation and injection of 20 vs. 40 ml mepivacaine 10 mg/ml. AB - The goal of this prospective randomized study was to assess the extension of the "three-in-one" paravascular lumbar plexus block after femoral nerve stimulation and injection of 20 vs. 40 ml mepivacaine 10 mg/ml. Three-in-one blocks were achieved in 12 of 39 (31%) patients given 20 ml of 1% mepivacine (group 1), and 17 of 41 (41%) patients given 40 ml (Group 2) of the same solution (n.s.). The level of successful blockade at each nerve did not differ between groups. The femoral nerve was blocked in 92% vs. 93% of patients in groups 1 and 2, respectively; the obturator nerve in 62% vs. 78%; and the lateral cutaneous femoral nerve in 41% vs. 44%. We conclude that femoral nerve stimulation is effective in faciliating blockade in the femoral nerve but not the obturator or lateral cutaneous femoral nerve with the tested solution and volumes, and therefore not particularly effective for achieving complete 3-in-1 blockade. Within the clinically relevant range of 20-40 ml, the volume of mepivacaine 10 mg/ml does not appear to influence the extent of blockade. PMID- 7484033 TI - Effects of sevoflurane on respiratory activities in the phrenic nerve of decerebrate cats. AB - Although the depressive effect of sevoflurane on ventilation has been reported, its potency and mode of action on the neural respiratory activity is still unclear. Therefore, the effects of sevoflurane on the phrenic nerve discharge and the respiratory timing were compared with those of halothane. The efferent activity of the phrenic nerve was recorded from decerebrate, un-anesthetized and artificially ventilated cats, and its power spectrum was calculated. The inspiratory and expiratory periods were measured. Sevoflurane and halothane of the doses of 0.5-1.5 MAC were inhaled for 15 min. With 0.5 MAC, sevoflurane decreased the total power and two dominant spectral components of the high frequency oscillation and medium-frequency oscillation in the power spectrum. With the same MAC dose, halothane had a greater depressive effect in a normocapnic condition with the vagus nerves being intact. In a state of hypercapnia or after vagotomy, the effect of halothane was considerably attenuated whereas that of sevoflurane remained unaltered. Halothane increased the neural respiratory rate much more than sevoflurane in both normocapnic and hypercapnic states. Vagotomy significantly weakened the effect of halothane to increase the respiratory rate but did not modify the effect of sevoflurane. With 1.0-1.5 MAC, both anesthetics severely decreased the phrenic power spectra and the potency difference became indistinct. The present findings demonstrate that sevoflurane has a weaker depressive effect on the respiratory nerve discharge and a smaller effect on the neural respiratory rate than halothane when the effects of 0.5 MAC were compared.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7484034 TI - Anaphylactoid skin reactions after intravenous regional anaesthesia using 0.5% prilocaine with or without preservative--a double-blind study. AB - Methylparaben, the preservative of various local anaesthetic solutions, is a potential allergen. In a double-blind study, 0.5% prilocaine with (Citanest, n = 100) or without (n = 100) methylparaben were compared for the occurrence of skin reactions after intravenous regional anaesthesia of the arm in surgical patients. Skin reactions were registered after the deflation of the tourniquet cuff, and intradermal tests were performed with 0.5% prilocaine, 0.1% methylparaben and saline in all patients. Seventeen patients in the Citanest group and four patients in the methylparaben-free prilocaine group developed erythematous skin reactions in the exposed arm after deflation of the tourniquet cuff (P < 0.05, between the groups). The skin symptoms disappeared within an hour and were always restricted to the region which had been anaesthetised. None of the affected patients had positive intradermal tests. The observed skin reactions are probably non-IgE-mediated anaphylactoid reactions in which the presence of methylparaben in the local anaesthetic solution plays a major role. PMID- 7484035 TI - Differences in midazolam-induced breathing patterns in healthy volunteers. AB - Sex differences, and the influence of drug dosage and additional upper airway obstruction were studied in midazolam-induced breathing patterns and sedation in 30 healthy volunteers (8 males and 22 females). After administration of 0.1 mg.kg 1 midazolam (8 male and 8 female subjects), the rib cage (RC) motion increased in 6 males and the abdominal wall (ABD) motion and SaO2 decreased in all males. In contrast, the RC and ABD motions and SaO2 decreased in all females. Snoring and loss of consciousness occurred in 7 males and in 2 females. There were significant differences in the RC motion, SaO2, the incidence of snoring and the sedative state between male and female subjects. A bolus dose of 0.5 mg of flumazenil completely antagonized the sedative effect of midazolam and restored the breathing pattern, whereas it did not completely restore SaO2. A higher dose (0.2 mg.kg-1) of midazolam was administered to an additional 8 females. It caused a loss of consciousness in all subjects and increased the RC motion in only one subject. Partial obstruction of the nasal cavity was effected with cotton balls in the remaining 6 females who were sedated with 0.1 mg.kg-1 midazolam. An increase in the RC motion occurred similar to that observed in males. These findings suggest a difference in midazolam-induced sedation and breathing patterns between male and female subjects with midazolam administration on a mg.kg-1 basis. PMID- 7484037 TI - Perioperative monitoring of neuromuscular transmission using acceleromyography prevents residual neuromuscular block following pancuronium. AB - The frequency of postoperative residual neuromuscular block following the use of the long-acting non-depolarizing muscle relaxants is high, and manual evaluation of the response to nerve stimulation does not eliminate the problem. In this prospective and randomized study we evaluated the hypothesis that perioperative use of acceleromyography would allow for a more rational and precise administration of the long-acting muscle relaxant pancuronium resulting in a decrease in 1) the incidence and severity of postoperative residual neuromuscular block, 2) the amount of pancuronium used, and 3) the time from end of surgery to tracheal extubation. Forty adult patients were randomized into two groups, one managed without the use of a nerve stimulator, the other monitored using train-of four (TOF) nerve stimulation and acceleromyography. All patients were anaesthetized with diazepam, fentanyl, thiopentone, nitrous oxide, and in some patients halothane, and they all received pancuronium 0.08-0.1 mg kg-1 for tracheal intubation, and 1-2 mg for maintenance of neuromuscular block. Neostigmine 2.5 mg preceded by atropine 1 mg was administered for reversal. In the patients managed without a nerve stimulator, the trachea was extubated when the anaesthetist judged the neuromuscular function to have recovered adequately for upper airway protection and spontaneous ventilation. In patients monitored with acceleromyography, the trachea was extubated when the TOF ratio was above 0.70. In all 40 patients, TOF ratio was measured using mechanomyography immediately after tracheal extubation and the patients were evaluated for clinical signs of residual neuromuscular block.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7484036 TI - Evaluation of left ventricular diastolic function in IgE-mediated anaphylaxis in dogs. AB - Depressed left ventricular (LV) systolic function remains controversial in patients with anaphylaxis and anaphylactic animal models. However, LV diastolic function has not yet been investigated in either patients or animal models. Therefore, we assessed LV diastolic and systolic function in IgE-mediated anaphylaxis in dogs. Seven dogs sensitized to Ascaris suum antigen were studied. An aqueous extract of Ascaris suum antigen was administered over 30 s into the systemic circulation to induce anaphylaxis. To assess LV function, the time constant of the fall of isovolumic LV pressure (T), as an index of LV diastolic function, and the maximum rate of rise of LV pressure (dP/dtmax), as an index of LV systolic function, were measured for a period of 120 min after antigen challenge. T was prolonged during the period of 5 to 15 min; however, changes in T were not significantly different from prechallenge values. LV d}/dtmax significantly decreased to 0.49 +/- 0.12 times the prechallenge value 5 min after antigen challenge, and significantly increased during the period from 30 to 120 min. However, these changes in dP/dtmax almost paralleled the changes in LV end diastolic pressure, indicating that left ventricular systolic function was relatively well preserved. In conclusion, LV diastolic function, i.e., isovolumic relaxation, is little impaired in anaphylaxis, and LV systolic function is relatively well preserved during the early stage following the onset of anaphylaxis. PMID- 7484038 TI - Spinal anesthesia attenuates myocardial ischemia during coronary artery spasm induced by intraaortic methacholine in rats. AB - Coronary artery spasm is not rare in patients with coronary artery disease, but the influence of regional anesthesia on spasm-induced myocardial ischemia is not known. We investigated the effects of spinal anesthesia on myocardial ischemia during coronary artery spasm in rats, and compared these with the effects of an alpha- and beta-adrenergic antagonist, and an alpha-adrenergic agonist. An intraaortic catheter was inserted via the right internal carotid artery so that the tip of the catheter was placed near the coronary ostium. An intrathecal catheter was placed at lumbar level. Coronary spasm was induced by the intraaortic injection of methacholine, and we identified the thresholds of myocardial ischemia, defined as the dose of methacholine that induced ST-segment elevation. Subsequently, the thresholds were determined after spinal anesthesia, and after the intraaortic injection of phentolamine and propranolol. The thresholds of myocardial ischemia increased significantly after intrathecal bupivacaine. In contrast, the threshold did not change after the injection of phentolamine. The thresholds increased significantly after the injection of propranolol. Methoxamine significantly decreased the threshold of ischemia. These results demonstrated that spinal anesthesia attenuated myocardial ischemia during methacholine-induced coronary spasm. This effect was equivalent to that of propranolol. PMID- 7484039 TI - QT interval of the ECG, heart rate and arterial pressure during anaesthetic induction: comparative effects of alfentanil and esmolol. AB - In a double-blind study the effect of esmolol and alfentanil on the QT interval of the ECG corrected by the heart rate (QTc), heart rate and arterial pressure during anaesthetic induction was studied in 59 oxycodone- and atropine premedicated ASA class I-(II) patients with a mean age of 26 yr (range 15-50 yr). The patients were randomly allocated to one of the four groups: saline, esmolol 2 mg.kg-1, esmolol 3 mg.kg-1 or alfentanil 0.03 mg.kg-1. Both doses of esmolol prevented the prolongation of the QTc interval after thiopental and suxamethonium, but not after laryngoscopy and intubation. Alfentanil prevented the prolongation of the QTc interval following thiopental, suxamethonium and laryngoscopy but not after intubation. Esmolol did not prevent the increase in the heart rate and arterial pressure in response to laryngoscopy and intubation. No cardiovascular responses to laryngoscopy and intubation occurred in the patients treated with alfentanil. No cardiac arrhythmias occurred in the esmolol 3 mg.kg-1 group, whereas the frequency of ventricular ectopic beats was 40% in the saline group and 13-20% in the other groups. PMID- 7484040 TI - Propofol and isoflurane induced EEG burst suppression patterns in rabbits. AB - The aim of this study was to compare propofol produced EEG burst suppression with isoflurane produced burst suppression in rabbits and to see whether rabbits can serve as models in studying the effects of different anaesthetics on human EEG. We recorded EEG of eight rabbits anaesthetised with isoflurane and propofol. The isoflurane bursts had higher amplitude than propofol bursts (P < 0.005). Isoflurane bursts appeared on distinct DC-shifts while propofol bursts were on slow waves. The EEG patterns were, however, different from those seen in humans. Rabbits did not have the rhythms seen in humans. We conclude that rabbits can be used to study the EEG effects of anaesthetics, such as the timing properties and reactivity of burst suppression pattern. However, this model seems less promising in the study of rhythmic activity seen in human EEG during burst suppression. PMID- 7484041 TI - Evaluation of anaesthesias in malignant hyperthermia negative patients. AB - In vitro contracture test is still the most reliable method for diagnosing malignant hyperthermia susceptibility. In order to investigate the reliability of Malignant Hyperthermia Negative (MHN) results, a questionnaire was sent to 237 persons of whom 133 were consecutively investigated and found to be MHN, together with 104 of their children. In 17 anaesthetics with triggering agents in 7 MHN patients and 7 children of MHN patients, no signs of malignant hyperthermia were observed in any patient. This material is, however, too small to draw any conclusion about the reliability of MHN test results. This can only be done in a large multicentre study or a metaanalysis of several studies. PMID- 7484044 TI - Respiratory changes during treatment of postoperative pain with high dose transdermal fentanyl. AB - This study made a longterm (72 hours) evaluation of the efficacy and possible side-effects of transdermal delivery of fentanyl (TTS-system) for post-operative pain relief. The study was double-blind, placebo-controlled with either a TTS system delivering fentanyl 100 micrograms.h-1 and rescue analgesic on demand or a placebo system and analgesic on demand. Analgesic consumption, pain, general satisfaction, respiratory rate, and levels of SpO2 and tcCO2 (pulse oximetry and transcutaneous CO2 measuring) were evaluated. Recruitment was stopped after enrolment of 24 patients, on safety grounds. The Fentanyl group was more satisfied with postoperative pain relief (P = 0.008); they had a lower analgesic demand (P < 0.05) but also a lower respiratory rate (P < 0.05) and a higher level of tcCO2 23 hours after application (P < 0.05). There were three cases (25%) of increased PaCO2 (> 6.5 kPa) in the Placebo group but without low PaO2 levels, sedation or bradypnoea. Conversely, there were three cases (33%) in the Fentanyl group with bradypnoea (< 10 breaths/minute), two without influence on PaO2 or PaCO2, but one (no. 24) with bradypnoea, heavy sedation, a marked decrease in PaO2 (5.8 kPa) and increased PaCO2 (7.5 kPa). These findings terminated the study. The 100 micrograms transdermal fentanyl system is agreeable to the patients, but apparently too potent for routine postoperative pain relief due to a risk of respiratory depression. Respiratory frequency can not be relied upon as sole indicator of insufficient respiration. PMID- 7484042 TI - The influence of anaesthetic drugs on the lower oesophageal sphincter in propofol/nitrous oxide anaesthetized dogs. Pressure profilometry in an animal model. AB - The lower oesophageal sphincter (LES) is the main barrier against gastro oesophageal reflux. Various anaesthetic drugs have been reported to affect the LES pressure. In this study, the effects of cisapride, atropine, suxamethonium, vecuronium and pancuronium on the LES pressure of six mongrel dogs anaesthetized with propofol and nitrous oxide were investigated. By means of eight-channel pressure profilometry the LES pressure was measured in consecutive sessions before and after administration of each drug. Compared to basal values, atropine and suxamethonium significantly decreased LES pressure, pressure vector volume and sphincter length. Cisapride significantly increased all sphincter parameters, vecuronium significantly increased LES pressure and pressure vector volume while pancuronium had no significant effects. A significant decrease of the LES pressure and pressure vector volume was observed when nitrous oxide was omitted from the ventilation mixture. Three-dimensional imaging showed an asymmetric shape of the LES pressure which resulted from radial differences of the LES pressure. The results from this study show that both the anaesthetic animal model and the eight-channel pressure profilometry are useful procedures in studying the effects on sphincter function of different drugs during anaesthesia. PMID- 7484043 TI - A porcine model for evaluation of cerebral haemodynamics and metabolism during increased intracranial pressure. AB - In patients with severe head injuries raised intracranial pressure (ICP) constitutes the most important cause of mortality. Several new therapies for increased ICP have recently been suggested and it is of importance to study the physiological effects of these treatments in animal experiments during steady state conditions. A porcine model for evaluation of cerebral haemodynamics and metabolism during increased ICP is presented. Intracranial hypertension was induced by inflation of two tonometric gastric balloons placed extradurally covering a major part of the parietooccipital region bilaterally. The distribution of the blood flow supplied by the carotid artery used for the cerebral blood flow (CBF) measurements was studied by intraarterial (i.a.) injection of 99mTc-HMPAO. The measurements showed that following ligation of the external carotid and the occipital artery no accumulation of tracer substance occurred in extracranial tissues during normal or increased ICP. Cerebral physiological variables (CBF, Cavo2, and ICP) were measured 5, 20 and 60 min after induction of intracranial hypertension. The results confirm that the experimental situation gives a reproducible increase in ICP (25-28 mm Hg) and that the physiological variables remain stable during the period of intracranial hypertension. We conclude that the model simulates the effects of an acute intracranial focal mass and is well suited for the evaluation of different pharmacological therapies of increased ICP. PMID- 7484046 TI - Morphine/prilocaine combination for intravenous regional anesthesia. AB - In this study we aimed to investigate clinically whether morphine acts on the peripheral nervous system directly. Twenty adult patients, who were scheduled for upper extremity surgery under tourniquet, had intravenous regional anesthesia. They were divided into two groups: one group was given 30 ml of 1% prilocaine plus 10 ml of 0.9% sodium chloride (P group) and the other group was given 30 ml of 1% prilocaine plus 6 mg morphine sulfate in 10 ml of 0.9% sodium chloride (P/M group). The onset of analgesia and anesthesia and recovery from anesthesia and analgesia were investigated in both groups. In the P/M group, the onset of analgesia and anesthesia was statistically faster and the recovery from anesthesia and analgesia was statistically slower than in the P group (P < 0.05). We conclude that morphine is clinically effective in the peripheral nervous system at this dosage range. PMID- 7484045 TI - Disturbance of hepatocellular integrity associated with propofol anaesthesia in surgical patients. AB - Propofol anaesthesia has not been associated with any hepatic consequences. We used glutathione transferase Alpha (GSTA), a very sensitive indicator of hepatocellular integrity, to evaluate the effect of propofol on the liver. Total intravenous anaesthesia was induced and maintained with propofol without any supplements in 30 female patients undergoing breast surgery. Ten healthy female volunteers given the lipid vehicle of propofol served as controls. Serum GSTA concentration was measured with a sensitive time-resolved immunofluorometric assay. Total intravenous propofol anaesthesia was stable and postoperative nausea negligible. A significant increase in GSTA from 3.1 micrograms.l-1 (mean baseline) to 10.0 micrograms.l-1 (mean peak) was noted after propofol infusion, indicating subclinical disturbance in hepatocellular integrity. No change in aminotransferases and no clinical signs of hepatotoxicity were observed. A small increase in GSTA from 2.4 micrograms.l-1 (mean baseline) to 4.1 micrograms.l-1 (mean peak) was observed during lipid infusion. We detected a subclinical disturbance in hepatocellular integrity after propofol anaesthesia for breast surgery. The mechanisms of hepatocellular impairment are not clear but the lipid vehicle of propofol alone does not explain it. PMID- 7484047 TI - The use of intravenous nitroglycerin for emergency cervico-uterine relaxation. AB - A patient with a twin pregnancy was in preterm labour after 25 gestational weeks when, during vaginal delivery of the second twin, severe spasm of the cervix and fetal bradycardia ensued. Induction of general anesthesia did not relax the cervix. After bolus doses of nitroglycerin 100 + 50 micrograms i.v., prompt cervico-uterine relaxation was obtained allowing manual extraction of the baby. A short review of the literature and a summary of our experience in the administration of nitroglycerin i.v. in obstetrics are presented. PMID- 7484048 TI - Antiarrhythmic effects of magnesium sulphate. Report of three cases. AB - A wide variety of antiarrhythmic agents is used in treatment of both supraventricular and ventricular arrhythmias. Magnesium sulphate has previously been used mainly in the treatment of torsade de pointe arrhythmias but several reports show that this agent may be used in the treatment of arrhythmias of different aetiology. We describe 3 patients who exhibited arrhythmias affecting haemodynamic performance. Case #1 had a subarachnoid haemorrhage and developed a supraventricular tachycardia. In case #2, ventricular tachycardia appeared during the postoperative course after abdominal surgery. Case #3 experienced critical heart failure due to dilated cardiomyopathy and had an irregular heart rhythm with multiple ectopic beats. In all three cases the administration of intravenous magnesium sulphate was successful in treating the arrhythmias. Magnesium sulphate is an antiarrhythmic agent that is effective mainly in treatment of ventricular arrhythmias. The drug can also be employed as second-line treatment of supraventricular arrhythmias. PMID- 7484049 TI - The 10th European Anatomical Congress, with the participation of the American Association of Anatomists. Florence, September 17-21, 1995. Abstracts. PMID- 7484050 TI - Etiology and pathogenesis of the muscular dystrophies. AB - Despite intensive research efforts, the cause of the muscular dystrophies has remained elusive for many decades. In the late 1980s, major advances in molecular genetics have led to the discovery of the dystrophin gene and its protein product, dystrophin. Mutations in the dystrophin gene result in dystrophin deficiency, which constitutes the pathogenetic basis of Duchenne and Becker muscular dystrophy. This major breakthrough set the scene for the ongoing discovery of the molecular basis of the muscular dystrophies. Several muscular dystrophies are caused by mutations in specific genes, which lead to deficiency of structural proteins of the sarcolemma other than dystrophin and to enzymatic dysfunction. Other muscular dystrophies have been mapped to different chromosomal loci, and in most of them, the identification of the molecular defect is ongoing. The elucidation of the molecular basis of the muscular dystrophies has a major impact on the understanding of their pathogenetic mechanisms, their nosological classification, their diagnosis and prevention, and, for the first time, offers perspectives for the development of causal treatment strategies. PMID- 7484051 TI - Central nervous system disease in primary Sjogren's syndrome. AB - Primary Sjogren's syndrome is a systemic autoimmune disorder whose main characteristics are dryness of the eyes and mouth, caused by lymphocytic infiltration of the exocrine glands. Patients may also show signs of extraglandular involvement of lung, liver, kidney and vessel walls, as well as of central and peripheral nervous system, muscles and joints. We present a review of the literature on the involvement of the central nervous system in primary Sjogren's syndrome, which may mimic many other neurologic disorders, especially multiple sclerosis. Features that may help in distinguishing these two entities are presented. PMID- 7484052 TI - [Craniostenosis and craniofacial dysmorphia]. AB - The paper gives a brief historical survey of craniostenosis and the first operative approaches. The authors describe a classification of various types of cranio-facio-stenosis and facio-cranio-stenosis and present a detailed study of their experience in this area. Operative results are discussed in function of the selected surgical procedure, the immediate per- and post-operative complications and the short and long term morbidity. The authors present their current attitude for each dysmorphic type. They underline the heavy nature of this long-lasting and bleeding surgical procedure that concerns small children. They have recorded 31 incidents in relation with surgical procedure, 10 complications at short and long term, and 1 death. Such surgery requires an experienced and vigilant team, associating surgeons, anesthetists and intensive-care physicians. PMID- 7484053 TI - Cerebral blood flow in just detoxified alcohol dependent patients. A 99 m Tc HMPAO-SPECT study. AB - Chronic alcohol abuse is responsible for several organic brain disorders. However, even the most characteristic of them are largely underdiagnosed by routine procedures. Therefore, there is need for sensitive, noninvasive and low cost diagnostic procedures. 99m-Tc-HMPAO-SPECT could be an interesting candidate in this indication, because it estimates the distribution of the cerebral blood flow and the metabolic activity of the brain with a good resolution. We used this technique in 17 healthy volunteers and in a sample of 50 patients dependent on alcohol, without other major physical or mental disorder. SPECT was performed during the attendance of these patients in an inpatient detoxification program. We observed abnormal SPECT in 34 patients, but only in 2 volunteers (p < 0.001). The main abnormality was heterogeneity of the distribution of the tracer. SPECT abnormalities appear to be due nor to withdrawal syndrome, nor to medication. On the contrary, they are correlated with stigmata of heavy alcoholism. Furthermore, a genetic vulnerability to alcohol was suspected because SPECT abnormalities are more frequent in patients with an history of drinking problems in their relatives. PMID- 7484054 TI - Association between stroke and atrial septal aneurysm assessed by transesophageal echocardiography in a cardiologic population. AB - Atrial septal aneurysm (ASA) is now recognized as a source of emboli. Its prevalence in a general population remains controversial. We assessed, in a cardiologic population, the prevalence of ASA, the association with cerebrovascular events, the role of associated interatrial shunting and age. Four hundred and ninety-four consecutive patients were referred for transesophageal echocardiography with contrast. The prevalence of ASA in two groups, with (A) and without (B) cerebrovascular events were compared. In group A, we studied the prevalence of ASA according to the clinical presentation (stroke, TIA and peripheral emboli in limbs). An ASA was found in 38 patients (8%). The prevalence of ASA was higher in group A (16%) than in group B (4%) (p < 10(-5)). Prevalence of ASA was not significantly different in stroke, TIA and peripheral emboli subgroups. There was no difference in prevalence of ASA in stroke or TIA subgroups according whether age was < 55 years or > or = 55 years. Prevalence of all types of vascular events was increased when ASA was associated with an interatrial shunt. Our results support the assumption that ASA is a potential source of cerebrovascular events, whatever the clinical presentation or age. Presence of interatrial shunt increases the risk of stroke in subjects with ASA. PMID- 7484055 TI - Target-like and comedon-like foci as different aspects of multiple healed brain abscesses. PMID- 7484058 TI - Infantile autosomal dominant distal myopathy. AB - INTRODUCTION: Distal myopathies are currently regarded as a non-homogeneous group of disorders including different autosomal dominant, recessive and sporadic forms. MATERIAL AND METHODS: The cases of a mother and her son and daughter are described and compared to previously reported cases from 4 families. Despite minor differences, the clinical picture is remarkably homogeneous, both within the same family and among different families. CONCLUSION: A distinct clinical form can be identified including: a) autosomal dominant inheritance; b) onset in infancy or childhood with peroneal muscles weakness; c) not disabling evolution in spite of possible late involvement of muscles others than tibio-peroneal; d) usually normal serum CK and other muscle enzymes; e) EMG evidence of primary myogenic damage; f) morphological findings of non-specific myopathy. Because of the benign evolution and the absence of true dystrophic changes in most biopsies we suggest the term infantile autosomal dominant distal myopathy should be preferred to infantile autosomal dominant distal muscular dystrophy. PMID- 7484056 TI - The increase of Parkinson's disease mortality could be due to a cohort effect. AB - INTRODUCTION: An increase of Parkinson's disease (PD) mortality has been observed in various countries during the last 3 decades. An analysis was done in order to define whether this trend could be due to a cohort effect. MATERIAL & METHODS: The mortality from PD in Italy during the period 1963-1987 was analyzed by means of a simplified cohort method. A mortality rate ratio (MRR) was also calculated, reporting all rates to 1906-1910 birth-cohort. RESULTS: The mean annual mortality rates from PD sharply increased during the period of the study. The MRRs increased from the cohort born between 1881 and 1885, to a maximum in the cohort born between 1906 and 1910, and then markedly decreased in the following cohorts. CONCLUSION: According to our findings, the increase of PD mortality could be partly ascribed to an higher risk of dying from PD experienced by the cohorts of people born in the first decade of this century. PMID- 7484057 TI - CSF and plasma concentrations of free norepinephrine, dopamine, 3,4 dihydroxyphenylacetic acid (DOPAC), 3,4-dihydroxyphenylalanine (DOPA), and epinephrine in Parkinson's disease. AB - OBJECTIVE: To investigate endogenous cerebrospinal fluid catecholamines in Parkinson's disease. MATERIAL AND METHODS: Basal concentrations of free norepinephrine (NE), dopamine (DA), epinephrine (E), 3,4-dihydroxyphenylacetic acid (DOPAC) and 3,4-dihydroxyphenylalanine (DOPA) in cerebrospinal fluid (csf) and plasma were measured using reverse-phase HPLC with electrochemical detection in 16 patients with Parkinson's disease and 21 control patients with low back pain. RESULTS: Parkinsonian patients had significantly decreased values of csf NE and DOPAC, the strong relationship between plasma and csf NE was disrupted and neither was there any age related increase of plasma NE. In l-DOPA treated patients plasma DA and DOPA concentrations were raised and csf DOPAC values were inversely related to severity of disease (Hoehn and Yahr score). Csf E concentrations were also reduced in parkinsonian patients whereas csf DA concentrations were unchanged. Csf DOPA concentrations were insignificantly decreased in parkinsonian patients. CONCLUSIONS: These results point towards a diffuse neuronal dysfunction in Parkinson's disease and indicate that lumbar csf NE and csf DOPAC are of central nervous origin. PMID- 7484060 TI - Expansion of a (CAG)n repeat region in a sporadic case of HD. AB - The genetic mutation underlying Huntington's disease (HD) has been identified as an expansion and instability of a specific CAG repeat sequence in a gene on chromosome 4. A simple polymerase chain reaction assay has been used for the assessment of the (CAG)n expansion in a 72-year-old woman with typical HD symptoms, but no family history of the disorder. The DNA analysis showed that the patient had an allele with 41 repeat units, in the size range seen in HD chromosomes. Therefore, HD diagnosis is confirmed in this seemingly sporadic case and the disease is newly diagnosed in a large family. The risk of inheriting this unstable expanded allele is discussed. INTRODUCTION--The discovery of an expansion of a trinucleotide (CAG) repeat region in the IT15 gene on the short arm of chromosome 4 has identified the mutational mechanism causing Huntington's disease (HD) and enables the direct diagnosis of affected subjects based on DNA analysis alone. Here a 72-year-old woman with typical HD symptoms, but no family history of the disorder, has been unambiguously diagnosed by using a quick DNA analysis. This is relevant because the disease is newly diagnosed in a large family. MATERIAL AND METHODS--A labelled polymerase chain reaction (PCR) test has been used to amplify the repeat region of the IT15 gene and DNA fragments were analyzed by Polyacrylamide gel electrophoresis. RESULTS--The number the CAG repeats in the proband displayed two alleles of 23 and 41 repeats, respectively. Since normal chromosomes are reported to contain 11-34 repeats, the clinical appearance of HD in the proband is explained by the presence of the repeat expansion. DISCUSSION--The parents of the proposita both died aged over 80 y apparently without neurological signs referable to HD. Hence, this is presumably a sporadic case of the disease. Because of the length of 41 repeats of this HD chromosome, offspring of this proband could inherit the expanded allele with 37 repeats, as expected for the reversal of the trinucleotide expansion. A subject with this intermediate allele could be affected, but would not be affected if the HD IT gene with reduced triplets had recovered its normal function. Thus, in a seemingly sporadic case like the one reported here, despite the PCR analysis, the risk of transmission of HD to her offspring may remain uncertain. PMID- 7484059 TI - Histopathologic and MRI findings in hypokalemic myopathy induced by glycyrrhizin. AB - Histopathologic studies and magnetic resonance images of the leg muscles were conducted in two patients with glycyrrhizin-induced hypokalemic myopathy (GHM). Muscle biopsy showed myopathic changes and vacuolated fibers with light microscopy, and dilatation of the sarcoplasmic reticulum, various types of vacuoles and myofibrillar degeneration with electron microscopy. High signal intensities in T2-weighted images obtained during severe muscle weakness were widely distributed in the leg muscles, especially the pretibial and soleus muscles. These high signal intensities disappeared after full recovery of muscle weakness. We suggest that high signal intensity in T2-weighted images can be seen and correspond to histopathologic changes in the muscles of GHM patients. PMID- 7484061 TI - Posatirelin for the treatment of late-onset Alzheimer's disease: a double-blind multicentre study vs citicoline and ascorbic acid. AB - INTRODUCTION: Posatirelin (L-pyro-2-aminoadipyl-L-leucil-L-prolinamide) a new synthetic tripeptide with cholinergic, catecholaminergic and neurotrophic properties, was investigated in the treatment of Alzheimer's disease. MATERIAL AND METHODS: A multicentre, double-blind study vs citicoline (reference drug) and ascorbic acid (inactive drug) was carried out in elderly patients suffering from late-onset Alzheimer's disease. The once daily intramuscular treatment lasted for three months followed by one-month oral treatment with a placebo. Subscales and factors of GBS (Gottfries-Brane-Steen) Rating Scale were identified as primary measures for efficacy assessment. RESULTS: At the end of the treatment, GBS subscale and factor scores assessing intellectual and emotional impairments, orientation and memory, ability to perform activities of daily living, depression anxiety, attention and motivation were improved in the postatirelin group, showing significant differences with respect to the citicoline and/or ascorbic acid groups. Tolerability was good in all groups. CONCLUSIONS: The improvement in the GBS Rating Scale score observed in the posatirelin group is clinically relevant. It is worth verifying the drug-induced functional improvements, in further studies with large samples. PMID- 7484062 TI - Electrically induced blink reflex and clinical blinking ability in patients with amyotrophic lateral sclerosis. AB - A study of the electrically induced blink reflex was performed on 17 patients with amyotrophic lateral sclerosis (ALS), graded according to the voluntary and reflexive blinking ability. In the early stage, delay of bilateral R2 latencies, decrease of bilateral R2 amplitudes and disappearance of the contralateral R2 component occurred. Then, the ipsilateral R2 component vanished when the voluntary blinking was incomplete. In cases with the remaining R2 component, the recovery curves of R2 amplitude revealed the weakened influence of the central nervous system to the interneurons that constructed the R2 component. In highly impaired patients who had lost the reflexive blinking ability, the R1 component had disappeared. Electrically induced blink reflex is useful in assessing the degree of disability in the upper facial muscles. PMID- 7484063 TI - Improving the diagnostic accuracy of the Mini-Mental State Examination. AB - INTRODUCTION: We determined the diagnostic accuracy of the Mini-Mental State Examination (MMSE) for dementia of the Alzheimer type (DAT) in an outpatient geriatric referral center in Switzerland. MATERIAL & METHODS: DAT patients and elderly controls were assigned to two groups: a validation sample (70 DAT patients; 50 controls) and a cross-validation sample (133 DAT patients; 43 controls). A Receiver Operating Characteristic curve was generated to derive the optimal MMSE cut-off score in the validation sample. RESULTS: The optimal MMSE cut-off was < 26/30 (sensitivity of 74%, specificity of 100%). Adjustments for age and education were necessary. The cross-validation confirmed these findings. CONCLUSION: iN A clinical setting the MMSE cut-off should be increased to < 26/30. A thorough neurobehavioral assessment is still necessary for a complete evaluation. PMID- 7484064 TI - White matter hyperintensity in neurologically asymptomatic subjects. AB - Recent advances in magnetic resonance imaging (MRI) technology have had a great impact on the delectability of minute, asymptomatic lesions of the central nervous system. The clinical significance and treatment modes of these lesions, such as white matter hyperintensity (WMH) lesions detected by T2 MRI, remain controversial. To address these problems, we retrospectively evaluated WMH lesions in relation to clinical parameters for 240 neurologically asymptomatic persons who had visited a hospital for a medical check-up of the brain. Proton and T2-weighted MRI were obtained using a 0.5 T superconducting MR imager using the spin echo technique with a repetition time (TR) of 2800 msec. An echo delay times (TE) of 40 msec was used for the proton MRI, and a TE of 100 msec was used for the T2-weighted MRI. The images were visually analyzed according to a four point grading system. The MRI findings were correlated with clinical parameters including age, gender, presenting symptoms, and hypertension. The overall frequency of WMH increased with age. Grades 2 and 3 of WMH were more frequent in aged persons, whereas the occurrence of grade 1 WMH remained relatively constant across age groups. Based on multiple regression analysis, age was the most significant variable influencing the frequency of WMH, followed by hypertension. These results imply that WMH lesions may simply be a phenomenon of aging, or may be an indicator of prepathologic state in an ischemic brain. PMID- 7484065 TI - Visual disturbances in patients with pituitary adenoma. AB - INTRODUCTION: To clarify the relationship between visual disturbances caused by pituitary adenomas and the degree of chiasmal compression, MRI (1.5T) findings were assessed two dimensionally. MATERIAL AND METHODS: 50 cases of pituitary adenomas and 17 control cases were studied. The position of the optic chiasm was measured by the distance from the line of the frontal base and posterior clinoid process on the sagittal T1 weighted image, and from the upper surface of the bilateral internal carotid artery on the coronal T1 image. RESULT: Visual disturbances appeared when the chiasm was displaced to more than 8 mm above the aforesaid reference level on the sagittal image and more than 13 mm above on the coronal image. There was a good correlation between the appearance of visual field defects and the degree of chiasmal compression. CONCLUSION: The degree of chiasmal displacement measured by MRI two dimensionally can provide useful information for predicting the appearance of visual disturbances. PMID- 7484066 TI - Rhabdomyolysis and hyperthermia after cocaine abuse: a variant of the neuroleptic malignant syndrome? AB - Rhabomyolysis with myoglobinuria has been added relatively recently to the neurologic complications associated with the increased use of cocaine and the introduction of its alkaloid form (crack). This retrospective study reports our experience with 14 patients who presented with rhabdomyolysis after cocaine use in a municipal hospital over a 3-year period. Seven patients used "crack", 2 intravenous and 3 nasal insufflation. All patients but one had hyperthermia, 11 altered mental status, 8 tachycardia, and 4 muscle rigidity. Nine developed renal failure; 3 of these patients died. Two other patients died of cardiorespiratory arrest. Cocaine-related rhabdomyolysis has a high mortality. The observed association with hyperthermia and other central neurologic features resembles the neuroleptic malignant syndrome. Since chronic cocaine use may alter the availability of dopamine either through transmitter depletion or decrease in the number of dopamine receptors, a common pathogenetic mechanism is possible. However, other mechanisms, which are not mutually exclusive but rather frequently overlapping, may play an important role. These include agitation, hyperthermia, adrenergic overstimulation leading to vasoconstriction and ischemia or calcium release from the sarcoplasmic reticulum resulting in increased entry into the muscle cell leading to cell death; in addition, cocaine has direct toxic effect on the muscles. PMID- 7484068 TI - Detection of aortic plaques by transesophageal echocardiography in patients with ischemic stroke. AB - Twenty-eight consecutive patients, aged 34-78 years with non-cardiogenic ischemic stroke were evaluated by transesophageal echocardiography (TEE). All patients were in sinus rhythm. Six of 28 patients (21.4%) displayed protruding masses in the aortic lumen. Five of these masses were located in the ascending aorta and one in the thoracic aorta. Our study suggests that cerebral infarction may also be due to aortic atherosclerotic plaques. Although our findings do not necessarily provide a causative link between atherosclerotic lesions in the aortic lumen and cerebral infarction, they may be an alternative potential source of stroke. TEE is the method of choice in detecting such lesions at the present time. PMID- 7484067 TI - Cerebrovascular CO2 reactivity in migraine with aura and without aura. A transcranial Doppler study. AB - INTRODUCTION: We studied by means of Transcranial Doppler (TCD) recordings the CO2 cerebrovascular reactivity in migraine patients during the headache-free period. MATERIAL & METHODS: In three groups of subjects (15 controls, 15 suffering from migraine with aura and 15 from migraine without aura) the middle cerebral artery (MCA) mean flow velocity (MFV) was recorded under basal condition and hypocapnia induced by hyperventilation. Relative MFV, PI (Pulsatility Index) changes and Reactivity Index (RI) were calculated. RESULTS: Reactivity Index values were: 0.019 +/- 0.007 (mean +/- SD) in control subjects: 0.029 +/- 0.008 in migraine with aura; 0.022 +/- 0.008 in migraine without aura. Statistical analysis showed a significantly (P < 0.05) increased RI in migraine with aura group. CONCLUSION: Cerebrosvascular CO2 reactivity is increased during the interictal period in migraine with aura patients. PMID- 7484069 TI - Doppler study of the ophthalmic artery in patients with carotid occlusive disease. AB - Assessing the adequacy of collateral circulatory pathways has become increasingly important in the investigation of cerebral circulation. Using transorbital Doppler ultrasonography (TOD), we examined the ophthalmic artery (OA) in patients with hemodynamic significant internal carotid artery (ICA) occlusive disease. The velocity and the direction of flow in the OA were studied in 45 patients (occlusion n = 27, stenosis > 75% n = 18), and in 30 age matched controls, under baseline conditions and within 30 minutes after the i.v. administration of 1 g acetazolamide as a vasodilatory stimulus. Based on the direction of flow before and after acetazolamide, the 45 patients could be separated into four groups with increasing degree of ICA lesions. In group I the OA flow was anterograde, but the velocity decreased after acetazolamide. In group II the OA direction became retrograde after acetazolamide, indicating collateral flow to the brain. In patients with retrograde OA flow following acetazolamide injection (group II-IV), the vessel reacted similarly to an intracranial artery, with marked increase in velocity when vasoreactivity was tested. TOD and the acetazolamide test provide useful information about potential collateral OA flow to the brain in patients with ICA occlusive disease. PMID- 7484071 TI - Non-convulsive status epilepticus as cause for focal neurological deficit. AB - In a four-and-a-half year period, 19 patients with focal neurological deficits accompanied by a focal electrographic status epilepticus were encountered. Sixteen of these patients showed clouding of consciousness or confusion. Computed tomography of the brain revealed focal lesions in 15 patients. In 7 patients the lesions were the result of a recent cerebral event and in 8 patients they were long-standing. All patients were treated with anti-epileptic drugs. Ten of the 12 patients without an acute lesion showed a complete recovery in a few days. In these patients the symptoms may have been caused by the continuous seizure activity, classifying them as cases of non-convulsive focal status epilepticus. Only 2 of the 7 patients with an acute lesion had a full recovery. In patients with an acute lesion the part played by the electrographic status epilepticus in the acquired deficits is unclear. Continuous or frequent intermittent focal epileptic discharges on the EEG may warrant treatment with anti-epileptic drugs in patients with focal neurological deficits, even when one of the hallmarks of epilepsy, clonic movements, is absent. PMID- 7484070 TI - Transitional progressive multiple sclerosis: MRI and MTI findings. AB - Transitional progressive multiple sclerosis (MS) is quite an unusual form of presentation and course of the disease. A case with this progressive form is presented and brain MRI and MTI findings are discussed in relation to the possible insight they may provide for understanding the mechanisms that determine progressive disability in MS. PMID- 7484072 TI - Short-term withdrawal of antiepileptic medication: effects on attention and memory. AB - INTRODUCTION: The aim of this study was to determine the effect of routine withdrawal of antiepileptic drugs (AEDs) during preoperative evaluation on cognitive functions and also to clarify whether short-term withdrawal of AED would help to lateralize the epileptic focus and to highlight focal dysfunction of memory in patients with temporal lobe epilepsy. MATERIAL & METHODS: Cognitive function was evaluated in 25 patients on full medication and after 1-week during the ongoing AED withdrawal. Six additional patients without AED withdrawal were studied during the same time interval and served as controls. The effect of AED withdrawal on memory tests was evaluated in a subgroup of 11 patients with temporal lobe epilepsy (TLE group). RESULTS: In the TLE group, verbal recurrent sequences learning tasks deteriorated significantly in patients with left-sided foci during AED withdrawal. In the whole group, attention, concentration and mental flexibility improved both in patients and in controls; the improvement was interpreted as being the result of the practice effect. During AED withdrawal, the number of errors in these tests decreased or remained unchanged. CONCLUSION: We suggest that preoperative neuropsychological testing can be scheduled at any time in patients referred for epilepsy surgery. In patients with temporal lobe epilepsy, the lateralizing results of selective memory tests may be most reliable if the testing is performed during AED withdrawal. PMID- 7484073 TI - Clinical characteristics of HTLV-1 associated dermato-polymyositis. Seven cases from Martinique. AB - Myositis linked to HTLV-1 is unfrequent. Over a period of 8 years, 14 patients with inflammatory myopathy were diagnosed in Martinique. Seven were seropositive for HTLV 1 antibody; the clinical and pathological data of whom are presented herein. Five patients presented with polymyositis, two with dermatomyositis. All seven patients had extra-muscular clinical features including neuropathy (4/7) and myelopathy (6/7), resulting in a quite peculiar clinical picture. Muscle biopsy showed a neurogenic process combined with myositic changes in 3/7 patients. Corticotherapy led to dramatic improvement in only one case, but with no sustained effect. HTLV 1 may be considered the etiological agent of this form of dermato-polymyositis, characterized by a clearly distinctive clinico pathological picture, and a poor response to corticotherapy. As in the case of tropical spastic paraparesis/HTLV 1 associated myelopathy, careful assessment of non-steroidal therapy is now warranted. PMID- 7484074 TI - Specificity of intrathecal IgG synthesis for HTLV-1 core and envelope proteins in HAM/TSP. AB - INTRODUCTION: In patients with human T-cell lymphotropic virus type 1 (HTLV-1) associated myelopathy/tropical spastic paraparesis (HAM/TSP), we investigated the significance of HTLV-1 specific antibodies in the cerebrospinal fluid (CSF). MATERIAL AND METHODS: The quantities of HTLV-1 specific immunoglobulin G (IgG) in paired CSF and serum were evaluated by a sensitive enzyme immunoassay (EIA). The specificity of antiviral IgG was determined by radioimmunoprecipitation of HTLV-1 antigens. RESULTS: In 17 of 20 HAM/TSP patients, quantitative evaluation by EIA supplied evidence for antiviral IgG synthesis within the CNS. Radioimmunoprecipitation demonstrated IgG antibodies against HTLV-1 envelope and core proteins in all HAM/TSP CSF and sera tested. Regarding the 3 sample pairs indeterminate in EIA for intrathecal synthesis, 2 showed stronger precipitation of HTLV-1 antigens by CSF IgG than by equal amounts of serum IgG. CONCLUSION: Intrathecal antibody synthesis specific for both HTLV-1 core and envelope antigens is common in HAM/TSP, thus providing conclusive evidence for an immune response to HTLV-1 within the CNS. PMID- 7484076 TI - Morphometric study of the frontal cortex in subacute sclerosing panencephalitis. AB - In biopsic material collected from the frontal cortex of 6 patients with subacute sclerosing panencephalitis (SSPE) and 5 patients with posterior fossa tumors, we estimated the neuronal and synaptic numerical densities as well as the mean volume of the neurons from layers II and III. The thickness of these layers was also determined. The evaluation of the layer's thickness suggested that there was no difference in the shrinkage in SSPE as compared to controls. No differences were found between the neuronal numerical densities and the neuronal soma sizes from SSPE and controls. Conversely the synaptic numerical density was reduced in SSPE. Given the maintenance of the neuronal numerical density in the frontal cortex of patients with SSPE, the presence of a decreased density of synapses must be regarded as a consequence of the dendritic and axonal degeneration that we previously described in this condition. It must then be borne in mind that SSPE's functional and behavioral changes might spring from alterations of the frontal cortex neuronal circuitry. PMID- 7484075 TI - Transcranial magnetic stimulation in multiple system and late onset cerebellar atrophies. AB - Central motor conduction time (CMCT) after transcranial magnetic stimulation (TMS) of the cortex, electromyography and nerve conduction velocity were performed in 24 patients with multiple system (MSA) and late onset cerebellar atrophy (LOCA) (often olivopontocerebellar atrophy--OPCA -). CMCT was abnormal in 7 patients with OPCA and one with LOCA. CMCT abnormalities (43% of cases) and increased threshold (68%) were more often found within OPCA group than in another multisystem atrophy and LOCA. Reduction in amplitude of the response after TMS was significantly correlated with cerebral hemispheres's atrophy. Increased threshold was correlated with upper vermal hemisphere atrophy and enlargement of the fourth ventricle. Electrophysiologic signs of mixed peripheral neuropathy were found in 8 patients. TMS abnormalities were not related to peripheral nerve involvement. Marked variation in CMCT suggests heterogeneity in these diseases. However, the percentage of CMCT abnormalities in OPCA group suggests that TMS seems to play a role in the neurophysiological diagnosis of these heterogeneous disorders. PMID- 7484077 TI - Ultrasound for the assessment of the embolic risk of carotid plaques. AB - In a prospective study we compared duplex-ultrasound characteristics of symptomatic internal carotid artery (ICA) stenoses with cranial computerized tomographic (CCT) findings in 82 patients suffering from completed or transient middle cerebral artery symptoms. The aim was to assess the pathogenic role of ICA plaque morphology and the potential embolic risk of ICA plaques. The degree of carotid stenosis was estimated by spectral analysis of the pulsed Doppler signal. The CCT findings were classified as being either normal, lacunar lesions, hemodynamically induced low-perfusion infarctions, or territorial embolic infarctions. According to their ultrasonic features we characterized the ICA plaque surface as smooth or irregular and their structure as homogeneous or heterogeneous. Plaques with an irregular surface and heterogeneous echogenicity dominated significantly in CCT-territorial infarctions (p < 0.01), whereas hemodynamically induced low-perfusion infarctions showed no relationship with any plaque characteristic. High degree ICA stenoses (> 50%) dominated in both territorial infarctions and low-perfusion infarctions, as compared to ipsilateral normal CCT or lacunes (p < 0.05). Normal CCT and lacunar infarctions were associated with homogeneous and smooth plaques (both p < 0.05). We conclude that > 50% ICA stenoses can cause both hemodynamically induced low-perfusion infarctions as well as thromboembolic territorial infarctions, whereas ulcerated and heterogeneous plaques constitute a high risk factor for arterio-arterial embolic stroke. Furthermore, carotid ultrasound may help to estimate the clinical significance of carotid lesions. PMID- 7484078 TI - Tomographic measurements of regional cerebral blood flow in progressive supranuclear palsy and Parkinson's disease. AB - Intellectual changes observed in progressive supranuclear palsy (PSP) are sometimes seen with lesser intensity in Parkinson's disease (PD). Cognitive impairment of PSP has been attributed to a frontal lobe dysfunction explaining the frontal cortex hypometabolism detected by PET. To establish whether this frontal hypometabolism is more pronounced in PSP than in PD, we compared frontal and temporo-parietal cerebral blood flow (CBF) indexes studied by SPECT using Tc99m HmPAO in 18 PSP, 18 PD and 8 control subjects. For each patient neuropsychological performances were also assessed. A significant left frontal hypoperfusion was observed in PSP (mean index value: 0.78 +/- 0.03, p < 0.01) and PD (0.78 +/- 0.04, p < 0.05) as compared to controls (0.84 +/- 0.03), whereas there was no difference between PSP and PD. No correlation was discovered between neuropsychological performances and frontal cortical index changes. This frontal uptake reduction of Tc99m HmPAO in PSP and PD could result from a disconnection phenomenon secondary to subcortical lesions. In both groups mean frontal indexes showed only a left frontal hypoperfusion suggesting that subcortical structures might be asymmetrically involved in early stages of the diseases. The lack of difference for indexe values between PSP and PD might be explained by the difference between the mean disease duration: 4.3 years for the PSP and 7.8 years for the PD. It might also suggest that frontal CBF reduction exists in the same proportions in PD and PSP, but at a later stage in the former case. PMID- 7484079 TI - Clinical significance of the ophthalmic artery in carotid artery disease. AB - A total of 141 subjects with tight stenosis (> or = 75%) or occlusion of internal carotid artery were followed up at intervals 3-6 months regularly for 40 +/- 16 months. The direction of ophthalmic artery flow was used as a parameter of risk indicator on cerebral ischemic events. Eleven patients with bilateral carotid tight stenosis/occlusion were excluded in the analysis. Thus, the 130 carotid arteries were divided into three groups: (1) carotid artery with ipsilateral hemispheric TIA or stroke (85 patients), (2) carotid arteries with contralateral hemispheric TIA/stroke or VBI (15 patients), and (3) carotid arteries of asymptomatic patients (30 patients). The symptomatic carotid artery group (group 1) had significantly more often reversed ophthalmic flow than the other two groups (group 2 and 3, p < 0.001). During follow-up prospectively for four years, 41 patients had cerebral ischemic events, three had cardiac ischemic events and six died of malignancy. Patients with reversed OA flow had more often subsequent cerebral ischemic events than those with forward flow (27 vs 14, p = 0.010). However, the difference remained significant only in the asymptomatic patients (group 3, 4 vs 0, P < 0.001), not for groups 1 and 2, after further analysis. Our work supported that the clinical role of ophthalmic artery collateral varied between asymptomatic and symptomatic patients. PMID- 7484080 TI - Cerebrospinal neuron-specific enolase, S-100 and myelin basic protein in neurological disorders. AB - In this study levels of neuron-specific enolase (NSE), S-100 protein (S-100) and myelin basic protein (MBP) in cerebrospinal fluid (CSF) of children and adults with distinct neurological disorders were examined. A previous study from our department demonstrated age related reference values for these brain-specific proteins in CSF. The median concentration level of the 3 proteins in 17 different neurological disease groups versus the reference group was compared. Significantly higher MBP values were observed in patients with multiple sclerosis (MS), cerebrovascular accident (CVA), metabolic disorder and infection. Furthermore, significantly higher values were demonstrated for S-100 in CVA and for NSE in metabolic diseases. In CVA, the NSE and S-100 values were significantly related with MBP values, whereas in MS the NSE and S-100 were not related with MBP values. PMID- 7484081 TI - CPEO and carnitine deficiency overlapping in MELAS syndrome. AB - Mitochondrial myopathy, encephalopathy with lactic acidosis and stroke-like episodes (MELAS) syndrome is one of the mitochondrial encephalomyopathies that has distinct clinical features including stroke-like episodes with migraine-like headache, nausea, vomiting, encephalopathy and lactic acidosis. We report a 27 year-old woman who presented with partial seizure, stroke-like episodes including hemiparesis, hemianopia and hemihypethesia, sensorineural hearing loss, migraine like headache, and lactic acidosis. Brain computed tomographic scan showed encephalomalacia in the right parieto-occipital area and recent hypodensity in the left temporoparieto-occipital area with cortical atrophy. Muscle biopsy revealed ragged-red fibers and paracrystaline inclusions in the mitochondria. Genetic study revealed an A to G point mutation at nucleotide position (np) 3243 of mitochondrial DNA. External ophthalmoplegia and ptosis were also found during two exaggerated episodes in this patient. Therefore, the overlapping syndrome of chronic progressive external ophthalmoplegia in the MELAS syndrome is considered in this case. Furthermore, we also found carnitine deficiency in this patient and she was responsive well to steroid therapy. Muscle biopsy also revealed excessive lipid droplets deposits. Therefore, the carnitine deficiency may occur in MELAS syndrome with the A to G point mutation at np 3243. We recommend the steroid or carnitine supplement therapy be applied to the MELAS syndrome with carnitine deficiency. PMID- 7484082 TI - Neurological outcome in coronary artery surgery with and without cardiopulmonary bypass. AB - INTRODUCTION: The neurological complications of coronary artery surgery with cardiopulmonary bypass (CPB) have been extensively studied, but to our knowledge those without CPB (NCPB) have not been defined. PATIENTS & METHODS: We prospectively examined 81 patients, before and up to seven days after surgery, to compare the neurological morbidity between patients subjected to coronary artery bypass graft (CABG) surgery with and without CPB. We analyzed demographic variables, risk factors and neurological examination including neuropsychological (NPS) tests using chi-square and non-parametric analysis (Mann-Whitney and Kruskal-Wallis). RESULTS: Forty-eight patients (34M; median age = 62 yrs; median number of grafts = 3 and median total surgery duration = 300 min) operated with CPB and 33 without CPB (23M; median age = 64 yrs; median number of grafts = 2 and median total surgery duration = 240 min) differed only in relation to number of grafts (p = 0.0001) and surgery duration (p = 0.0001). CONCLUSION: We found no difference in early neurological outcome in patients subjected to CABG with or without CPB. PMID- 7484083 TI - Respiratory function deterioration is not time-linked with upper-limb onset in amyotrophic lateral sclerosis. AB - INTRODUCTION: In amyotrophic lateral sclerosis (ALS), symptoms apparently spread following regional rules, and depending on the site of onset. We examined if respiratory function deterioration appears earlier or is more severe in patients with upper-limb onset. MATERIAL & METHODS: We compared the results of various pulmonary function tests (PFT) obtained at diagnosis depending on the site of onset in 49 ALS patients. In a longitudinal study, we compared the deterioration of forced vital capacity (FVC) in relation to the site of onset, and analyzed the time elapsed to reach values below 80% of predicted according to site of onset, and we compared the survival depending on the site of onset. RESULTS: No significant differences in PFT were found in the upper-limb onset group in any of the analysis performed. No differences in survival were detected in any disease onset group. CONCLUSION: Pulmonary function deterioration does not appear to be time-linked to arm onset in ALS. PMID- 7484084 TI - Mechanical and electromyographic stretch responses in spastic and healthy subjects. AB - The reflex mediated mechanical response was studied in the ankle flexors and ankle extensors of healthy and spastic subjects at maintained contractions from low to high concentration levels. This was done by a technique where muscle stretches could be applied during contractions with stretch reflex responses present or during contractions where the stretch reflex was absent. Stretch responses without stretch reflexes were obtained during contractions elicited by electrical stimulation. The validity of this method is discussed in details and it is concluded that the stretch responses during electrical stimulation can give a correct estimate of the non-reflex muscle response. The method is difficult to carry out in many human subjects and a number of precautions have to be taken. In healthy subjects a large reflex mediated mechanical response was found in the ankle flexors and ankle extensors, with the largest response at low and intermediate contraction levels. Surprisingly the reflex mediated mechanical response was found to be of equal size in the ankle extensors of spastic patients and control subjects at all contraction levels. In the ankle flexors no reflex mediated mechanical response was present in the patients contrary to the findings in the control subjects. A method was developed to predict the reflex mediated mechanical response from the reflex mediated EMG response. The method was successfully applied in the ankle flexors. In the ankle extensors the measured reflex mediated mechanical response was a factor of 2.5 lower than the EMG predicted mechanical reflex response. It was concluded that the method cannot be applied in situations where a large synchronized EMG response occurs--as it does in the ankle extensors. An increased EMG response was found in the ankle extensors in spastic patients, but this was not followed by an increased mechanical reflex response. This emphasizes that conclusions drawn from EMG results should be done with caution. Stretch reflexes are increased in spastic patients during clinical examination. This is in contrast to the findings under our experimental conditions, where the reflex mediated response during maintained contraction was decreased in the ankle flexors and unchanged in the ankle extensors of spastic patients. Others have found that the H-reflex is modulated in healthy subjects in relation to different motor tasks. It was proposed that healthy subjects set the reflex in a facilitated state in relation to ongoing contraction under our experimental conditions and perhaps in a more inhibited state in the clinical test situation.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 7484086 TI - 3.1-kb deletion of mitochondrial DNA in a patient with Kearns-Sayre syndrome. AB - Mitochondrial DNA (mtDNA) deletions have been found in the majority of patients with chronic progressive external ophthalmoplegia and Kearns-Sayre syndrome. A large number of different mtDNA deletions have been identified. They generally spare the two origins of replication and are frequently flanked by direct or indirect repeats. We have found a 3.1-kb deletion of mtDNA in a patient with Kearns-Sayre syndrome that has some unusual features. First, it encompasses nucleotides 11259 to 14368, a localization that was not described before. Second, the deletion is not flanked by direct or indirect repeats, supporting the view that homologous recombination and slip-replication do not account for all mtDNA deletions. PMID- 7484087 TI - The effect of MS-818, a pyrimidine compound, on the regeneration of peripheral nerve fibers of mice after a crush injury. AB - One of the pyrimidine compounds, 2-piperadino-6-methyl-5-oxo-5,6- dihydro(7H)pyrrolo[3,4-d]pyrimidine (MS-818), has neurotropic effects in vitro. Therefore, we studied the effect of MS-818 on the regeneration of the peroneal nerve in C57BL/6J mice after a crush injury. Two test groups, which received a daily intraperitoneal injection of 5 mg/kg or 10 mg/kg MS-818, respectively, were compared with controls, which received daily intraperitoneal injections of physiological saline, over a 14-day period. The maximum foot-width ratio (crushed side/uncrushed side) was obtained on days 1, 8 and 14 after the crush injury, and the various morphometric parameters were evaluated at both 5 and 10 mm distal to the proximal portion of the crush site. The significant effects of MS-818 included a larger maximum foot width (P < 0.04) and a greater number of unmyelinated axons per nerve at both levels (P < 0.003) in both test groups than in controls. MS-818 had no significant effects on body weight, the increase of total transverse fascicular area after the crush injury, the total number of myelinated fibers with their size distributions, or the number of nuclei of Schwann cells and macrophages. Therefore, we conclude that MS-818 promotes axonal sprouting and elongation after a crush injury in mice. PMID- 7484088 TI - Cerebral beta amyloid deposition in patients with malignant neoplasms: its prevalence with aging and effects of radiation therapy on vascular amyloid. AB - We examined immunohistochemically 123 autopsy brains from patients aged between 30 to 59, who died as a result of malignant neoplasms. Using antiserum to amyloid beta protein (A beta), we found that cerebral A beta deposits began in the subjects' fifth decade; its prevalence was 0%, 9.8% and 21.5% in the fourth, fifth and sixth decades, respectively. The major form of A beta deposition was diffuse-type plaques, although one third of the brains with A beta deposition showed amyloid angiopathy. Subpial A beta deposition is frequently associated with amyloid angiopathy. The prevalence of cerebral A beta deposits was about two times higher in the patients who had received brain radiation therapy (27.8%) compared to non-radiated patients (14.8%). Amyloid angiopathy was much more prominent (P < 0.05) with radiation therapy (22.2%) than without (8.0%). We found that cerebral A beta-deposition is dependent on aging, even in patients with malignant tumors and at beginning in their forties, and that brain radiation therapy is a possible risk factor of A beta deposition, especially in the form of amyloid angiopathy. PMID- 7484085 TI - Prions, beta-sheets and transmissible dementias: is there still something missing? PMID- 7484089 TI - Differential expression of triiodothyronine receptors in schwannoma and neurofibroma: role of Schwann cell-axon interaction. AB - Regulation of gene expression in Schwann cells may be determined, at least in part, by the interaction of these cells with axons. Two peripheral nerve tumors, neurofibroma and schwannoma, represent good tools for studying Schwann cell activity in the presence or absence of axon action. In the present work we studied the expression of triiodothyronine receptors (T3R) by Schwann cells in these two tumors and also in adult normal sciatic nerve. Confirming the results of the histological examination, immunostaining of the neurofilaments showed the presence of fascicles or scattered axons in all neurofibroma sections studied. In these neurofibromas, Schwann cells did not express T3R immunoreactivity. Furthermore, in adult normal sciatic nerve, Schwann cells which ensheathed axons were devoid of any T3R expression. In contrast, in schwannoma, the complete absence of axons was demonstrated by the lack of neurofilament immunostaining. Here, Schwann cells deprived of axonal interaction displayed clear T3R immunoreactivity. In schwannoma cell cultures, Schwann cells continued to express T3R, even in cultures treated with medium that had been conditioned with rat sensory neurons. On the basis of these results, we suggest that, beside the possible regulatory mechanisms for T3R, the synthesis of T3R is regulated, at least in part, by Schwann cell-axon interaction. PMID- 7484090 TI - Cytoplasmic body myopathy: familial cases with accumulation of desmin and dystrophin. An immunohistochemical, immunoelectron microscopic and biochemical study. AB - Muscle biopsy samples from five patients with cytoplasmic body myopathy (CBM) were investigated by immunohistochemical (antibodies to desmin, actin, dystrophin, spectrin, alpha actinin and utrophin), immunoelectron microscopic (antibodies to desmin, actin and dystrophin) and biochemical (desmin, dystrophin, actin and utrophin western blots) methods. Using immunofluorescence it was shown that the centers of cytoplasmic bodies (CB) were stained by anti-actin, anti utrophin and three different anti-dystrophin antibodies. The peripheries were labeled by the anti-desmin antibody. Moreover, fibers containing CB showed a markedly increased staining of their entire sarcoplasm with the anti-desmin antibody. Using immunoelectron microscopy it was shown that anti-dystrophin antibodies selectively stained the external limit of the central granular region. Anti-desmin antibody labeled the filamentous halo, and anti-actin antibody stained the central core and the radiating filaments. Biochemical studies showed storage of desmin and dystrophin, both of normal molecular weight. Our results suggest that CBM should be considered along with a wider group of intermediate filament pathologies that include desmin-storage myopathies. PMID- 7484092 TI - Monocyte recruitment into the scrapie-affected brain. AB - The recruitment of monocytes into the scrapie-affected brain was investigated in female mice reconstituted with male bone marrow, using a Y-chromosome-specific probe and F4/80 immunocytochemistry. Recruitment of monocytes could be demonstrated in six out of eight animals and the number of recruited cells correlated with the severity of vacuolation in most, but not all, animals. The proportion of microglia derived from recruited monocytes varied between individual animals, did not correlate with the increase in cellularity (glia) in affected areas of brain and did not affect the length of incubation period. Thus, it is unlikely that the recruitment of monocytes is a pivotal event in the development of early pathological changes in scrapie. The morphology of recruited cells in scrapie lesions, as revealed by F4/80 immunoreactivity, was indistinguishable from that of activated resident microglia. PMID- 7484091 TI - Peripheral nerve pathology in Niemann-Pick type C mouse. AB - The sciatic nerve of the mouse mutant with Niemann-Pick type C disease (NPC mouse) was investigated using light and electron microscopy, and teased-fiber preparations. As early as postnatal day 20, when clinical symptoms were not yet apparent, focal paranodal swellings with an accumulation of small myelin figures in the Schwann cell cytoplasm were noted. These paranodal changes were more pronounced in the distal segment and became progressively conspicuous with increasing age. The morphometric analysis revealed a hypomyelination of large myelinated fibers in the NPC nerves at 70 days, whereas an essentially similar histogram pattern was noted in both control and NPC nerves at 20 days, suggesting progressively defective utilization of cholesterol in the NPC nerves with age. Intraxonal accumulation of dense bodies was noted in older mice, but no segmental demyelination or Wallerian type of axonal degeneration was observed at any age. The changes noted in the paranodal regions in the NPC mouse closely resemble those found in rats treated with an inhibitor of cholesterol biosynthesis, as well as those seen in remodeling fibers during an early stage of peripheral nerve development. Thus, the morphological changes seen in the sciatic nerve of the NPC mouse may be an expression of perturbation in myelin maintenance as a result of defective cholesterol metabolism. PMID- 7484093 TI - Synaptic loss in the proximal axon of anterior horn neurons in motor neuron disease. AB - This report deals with an ultrastructural investigation of the synapses of the proximal axons of normal-appearing anterior horn neurons of 7 patients with amyotrophic lateral sclerosis (ALS) and 4 patients with motor neuron disease who had no upper motor neuron and corticospinal tract involvement (lower motor neuron disease, LMND). Specimens from 12 age-matched individuals who died of non neurological diseases served as controls. Proximal axons directly emanating from the normal-appearing neurons were examined: 42 axons were from ALS patients, 43 from LMND patients and 87 from controls. Our results show that the number of synapses on axon hillocks, as well as the lengths of the synaptic contact and of the active zone were reduced in both groups of patients (P < 0.0001), but no significant differences were seen between patients and controls with respect to the synaptic parameters of initial axon segments. There was no overall difference between ALS and LMND patients. These findings suggest that the electrophysiological functions pertaining to integration of electrical inputs into the axon and information transduction on the axon may be greatly impaired in the early stages of motor neuron diseases, and that the observed synaptic alterations may be pathological events, likely to be due to anterior horn neuron degeneration. PMID- 7484094 TI - Familial fetal akinesia deformation sequence with a skeletal muscle maturation defect. AB - Two female siblings with the fetal akinesia deformation sequence (FADS) are described. Both showed facial anomalies, arthrogrypotic extremities, hypoplastic lungs, and fetal growth retardation. The central nervous system of the second sibling, including the spinal cord, was normal. The skeletal muscle was studied by immunohistochemistry for the expression of several myosin heavy chain isoforms, M-band proteins and intermediate filament proteins. The skeletal muscle was immature and atypical muscle spindles containing up to 31 intrafusal fibers were found. These findings suggest that a lethal FADS phenotype may involve a maturation defect of the skeletal muscle, and, in this family, may be inherited in a recessive fashion. PMID- 7484095 TI - Immunoreactivity of the central nervous system in cats with a Borna disease-like meningoencephalomyelitis (staggering disease). AB - The inflammatory cell composition and the expression of major histocompatibility complex (MHC) antigens in the central nervous system (CNS) of 13 cats with a spontaneous, Borna disease-like meningoencephalomyelitis (staggering disease) was investigated by immunohistochemistry with a panel of monoclonal and polyclonal antibodies. T lymphocytes were the predominating inflammatory cells within the adventitial space. CD4+ T cells were more abundant than CD8+ T cells. Scattered IgG-, IgA- and IgM-containing cells were found in the adventitial space and surrounding neuropil, often adjacent to neurons. There was a markedly increased MHC class II expression in cells morphologically resembling microglia. In several cats, Borna disease virus specific antigen was detected, but only in a few cells, mainly of macrophage character. Our findings indicate a long-standing inflammatory reaction in the CNS of cats with staggering disease, possibly triggered and sustained by a persistent viral infection. PMID- 7484096 TI - Alzheimer paired helical filaments: a comparison with the twisted ribbon model. AB - To investigate if Alzheimer paired helical filaments (PHF) closely resemble twisted ribbons, as indicated by recent high-resolution ultrastructural studies, we compared physical models of twisted ribbons with electron microscopic images of PHF. Uranyl-acetate-stained, isolated PHF with one or two helical turns were compared with scale models of twisted ribbons with one and two helical turns rotated at different angles. The various rotations of the twisted ribbon model corresponded well with the different orientations of randomly dispersed PHF. The electron-dense regions of individual PHF turns previously thought to represent a cross-over site of paired filaments corresponded to the edge of the twisted ribbon when the ribbon was oriented perpendicular to the filament axis. These data indicate that the overall configuration of PHF is a twisted ribbon but does not exclude possible configuration restrictions due to an ordered arrangement of subunits. PMID- 7484097 TI - Cerebellar superoxide dismutase expression in Menkes' kinky hair disease: an immunohistochemical investigation. AB - This comparative immunohistochemical study deals with the expression of the cytosolic Cu/Zn-binding and mitochondrial Mn-dependent superoxide dismutases (SODs) in the cerebella of five patients with Menkes' kinky hair disease (MKHD) and five age-matched controls. Several cell types, including Purkinje cells and reactive astrocytes, of all MKHD patients examined were intensely stained by an antibody to Mn SOD, but not by an anti-Cu/Zn SOD antibody. By contrast, the cells of the five controls reacted very weakly or not at all with the anti-Mn SOD antibody, but were strongly reactive with the antibody to Cu/Zn SOD. These results suggest that the increased Mn SOD immunoreactivity in MKHD reflects enzyme induction as a protective mechanism against the highly toxic superoxide anion generated under the disease conditions. PMID- 7484098 TI - Polymerase chain reaction fiber analysis and somatic mosaicism in autopsied tissue from a man with Duchenne muscular dystrophy. AB - Single muscle fibers, obtained at autopsy from a 22-year-old man with Duchenne muscular dystrophy were examined immunocytochemically and also using polymerase chain reaction (PCR). Dystrophin-positive cells were widespread in skeletal, cardiac, smooth muscle, and in brain cells. PCR and Southern blot analyses of DNA from peripheral blood lymphocytes revealed a deletion of exon 45 in the dystrophin gene. With PCR of single fibers, three bands corresponding to exons 44, 45, and 47 were present in the normal control muscle fibers and dystrophin positive fibers from the patient, while only two bands, exons 44 and 47, were observed in dystrophin-negative fibers. Therefore, in this patient, the genotype of dystrophin-positive fibers differed from that of the dystrophin-negative fibers, possibly because of a somatic mosaicism for deletion in the dystrophin gene. A mutation of the dystrophin gene may have occurred in one cell at an early stage of ontogenesis. PMID- 7484099 TI - Clinical and neuropathological features of a neurodegenerative disorder in the central nervous system with progressive head drooping (Kubisagari). AB - The clinical and neuropathological features of a case of a neurodegenerative disorder with pronounced and progressive head drooping, in Japanese Kubisagari, are reported. This female patient died at the age of 72 years after an approximately 20-year history of peculiar posture with progressive head drooping (Kubisagari) and lordosis (bowed posture), parkinsonism, dysphonia and slight muscle wasting of the face, tongue, neck, and distal portions of the upper extremities. She did not display mental deterioration until the terminal stage of the illness. A simple macroscopic inspection formalin-fixed sections of the central nervous system (CNS) showed prominent atrophic frontal and temporal lobes, brownish discoloration of the putamen and an atrophic pyramidal tract. Light microscopy revealed severe neuron loss with fibrillary gliosis at both the above-mentioned lobes and the putamen. Both the facial and hypoglossal nuclei had almost disappeared. Motor neurons in the spinal cord were moderately to markedly decreased. Neither Bunina nor Lewy bodies, senile plaque, nor Pick's argyrophilic neuronal inclusions were observed, but very occasionally ubiquitin-positive neurons were found in the temporal cortex. In conclusion, the hitherto unrecognized neuropathological findings in the CNS corresponding to progressive head drooping (Kubisagari) suggest that this is a neurodegenerative disorder of the CNS, possibly an atypical form of amyotrophic lateral sclerosis. PMID- 7484100 TI - The neuropathology of cerebrotendinous xanthomatosis revisited: a case report and review of the literature. AB - Cerebrotendinous xanthomatosis (CTX), a rare autosomal-recessive lipid storage disease, has been well characterized clinically and biochemically, and recently also from the molecular biological aspect. However, only a very few publications deal with its neuropathology, and views on its pathogenesis vary. Based on a recently examined case, we propose that central-peripheral distal axonopathy is the major pathogenetic mechanism of nervous system injury in CTX. The latter is characterized by white matter pathology, typically in form of long tract involvement with the more distal parts of the tract more severely affected. Most severely affected are the cerebellar white matter, the optic pathways and the long tracts of the brain stem and spinal cord, particularly the pyramidal tracts, although there is hardly a CNS region which does not display some form of pathology. Lesions are characterized by loss of myelinated fibers and accumulation of lipid products in form of foamy macrophages, clear oil-red-O positive spaces and crystalline clefts, accompanied by gliosis, occasional axonal spheroids, and in the cerebellum--the most severely affected structure--also by multi-nucleated foreign body giant cells. Demyelination is not seen, and ultrastructurally myelin sheaths are normally structured. Signs of axonal degeneration are also present in the spinal roots. We hypothesize that the basic enzymatic defect in CTX leads to accumulation of metabolites in the brain which may be neurotoxic and may impair the metabolic apparatus of neurons with resultant axonopathy and subsequent nonspecific lipid deposition in the injured tracts. PMID- 7484101 TI - Computerized measurements of the lower third molar related to chronologic age in young adults. AB - Chronologic age is an utmost important birth record in many industrialized Western countries. When the date of birth is unknown, there is a demand for establishing the chronologic age. Dental methods for age estimations are considered more reliable than most other methods by the majority of researchers and are among the most commonly used means to determine age. Traditionally, these methods have most often been based on subjectively estimated, radiographic stages of tooth development. In the present study metric measurements of the length of the forming root have been used and are compared with one standard method. The formed part of the root of the lower third molar as seen in a panoramic radiograph was digitized. Data were directly stored in a computer for subsequent statistical analysis. The technique of collecting data with the aid of a computer had some statistical advantages. The precision of age estimations was, however, slightly inferior to the standard method. PMID- 7484104 TI - Mouth-rinsing with chlorhexidine causes a delayed, temporary increase in the levels of oral viridans streptococci. AB - The indigenous oral flora of 27 volunteers was monitored longitudinally over a 4 week period. Bacteria attached on buccal epithelial cells were counted by microscopy. Salivary bacterial colonies and the presence of alpha-hemolysis were examined after aerobic culturing on blood agar plates. The buccal and salivary bacterial counts were stably maintained in most subjects in the two repeated base line samplings taken at 1-week intervals. Rinsing with a chlorhexidine mouthwash 45 min before sampling dramatically reduced the amount of epithelial cell adherent bacteria. One day after the chlorhexidine rinse, however, the numbers of the epithelial cell-adherent bacteria exceeded the base-line level, and a similar decrease-increase pattern of changes was detected for the salivary alpha hemolytic streptococcal counts. The non-hemolytic salivary bacterial counts were not affected by chlorhexidine. Subsequent weekly samplings showed no difference from the base-line samplings. The chlorhexidine-induced, delayed increase of viridans streptococci on oral epithelial surfaces should be considered a possible risk factor in medically compromised patients. PMID- 7484102 TI - Cavity design and dimensions of tunnel preparations versus composite resin Class II preparations. AB - Fourteen pairs of extracted contralateral premolars with small, artificial, standardized, approximal 'caries lesions' were placed in 14 plaster jaws. Fourteen dentists made a mesial tunnel preparation and a distal composite resin preparation on one tooth and vice versa on the contralateral tooth. Less tooth substance was removed in the tunnel preparations than in the class-II preparations, but this difference was not statistically significant when the resin class-II preparations were made without occlusal retention. Twenty-five per cent of the tunnel preparations had residual 'caries', as opposed to 7% in the class-II composite preparations. Tunnel preparations with larger occlusal openings had less residual 'caries'. The morphology of the class-II resin preparations varied considerably, indicating a lack of precise descriptions in the dental literature. PMID- 7484103 TI - Resin-modified glass ionomer cements: fluoride release and uptake. AB - The aim was to study the short- and long-term fluoride release from resin modified glass ionomer cements (GIC). The aim was also to determine the effect of fluoride treatment of 9-month-old specimens, consistency of the mix, and pH of the environment on the fluoride release. GIC test specimens were continually exposed to running water, and the fluoride release was measured periodically by storing the specimens in 5 ml deionized water for 1 week and measuring the fluoride content of the solution. After 24 h, 1 month, 9 months, and 11 months in running water four of the six resin-modified GICs released as much as or more fluoride than the auto-curing GIC tested for comparison. Fluoride treatment after 9 months also increased the fluoride release of these four brands, as was the case with the conventional GIC. At 24 h and 1 month two of the resin-modified GICs released smaller amounts of fluoride than the other materials, and the fluoride treatment used on those had no or only a minimal effect. Thin consistency of a mix resulted in higher fluoride release for one resin-modified material than a thick mix. Low pH increased the fluoride release for all materials. PMID- 7484105 TI - Questionnaire survey on the use of dental X-ray film and equipment among general practitioners in the Swedish Public Dental Health Service. AB - A questionnaire was sent to all clinics in the Public Dental Health Service in Orebro County in 1989 and 1993. The survey comprised questions about X-ray film and developing technique. The questionnaire of 1993 was accompanied by questions on the type of dental X-ray film and collimator each dentist used, altogether 175 dentists. The results showed that Kodak Ektaspeed dental X-ray film was used by 53% of the dentists in 1993. The mean number of months to the expiry date for all film packages irrespective of film type increased from a mean of 7.5 months to a mean of 13 months from 1989 to 1993. Only 11% of the Ektaspeed film packages in 1989 had a base plus fog < or = 0.25, whereas the figure in 1993 was 30%. Rectangular collimation adjusted to the size-2 film (31 mm x 41 mm) was used by 36% of the dentists in 1993. In 1993, 88% of the clinics used automatic processing, and in 85% of the clinics both automatic and manual processing was used. The processing time and temperature varied greatly for both automatic and manual processing. The results indicate that film and developing procedures in Swedish general dental practice are not always in accordance with the recommendations and guidelines of the Swedish National Radiation Protection Institute (SSI) and that attempts should be made to improve dentists' behavior with regard to radiation safety. A major dose reduction would be achievable without jeopardizing diagnosis if the regulations of the SSI were followed by using the fastest available film and optimum developing procedures.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7484106 TI - Psychologic aspects of patients with oral lichenoid reactions. AB - Psychologic aspects of 49 patients with oral lichenoid reactions (OLR) in contact with amalgam fillings were studied and compared with an age- and sex-matched control group. Psychologic factors such as personality, psychologic functioning, and quality of life were determined by using the Karolinska Scales of Personality (KSP), an additional Personality Scale (PS), a Psychological Functioning Scale (PFS), and a Quality of Life Scale (QLS). With regard to personality the OLR patients had significantly higher scores on the muscular tension and suspicion scales and significantly lower scores on the indirect aggression scale. In addition, the OLR patients were significantly more worried about their health and more helpful. With regard to psychologic functioning the OLR patients had significantly more sad thoughts, became dizzy more easily, found it harder to imagine themselves free from anxiety, and had more difficulty in concentrating. The results indicated that OLR patients had a tendency to be depressive. The need for a systemic investigation including odontologic, medical, and psychologic aspects was expressed. PMID- 7484108 TI - Fatty acid composition of palatal tissue from denture stomatitis patients. AB - Palatal biopsy specimens were obtained from patients with denture stomatitis. The fatty acids were extracted from the tissue, then separated, identified, and quantified by a gas-chromatographic technique. The sensitivity of this method enabled analyses of specimens with a wet weight of less than 1 mg. The concentration of the fatty acids C16:1 (n-7) and C24:1 (n-9) differed significantly between samples from hyperplastic and clinically healthy tissue in the denture stomatitis patients. By comparing specimens from denture stomatitis patients and non-denture subjects, the concentration of seven fatty acids, two saturated and five unsaturated, was found to be significantly different. A multivariate data-analytical method distinguished between the fatty acid composition in specimens from denture stomatitis patients and from non-denture subjects. PMID- 7484109 TI - Maximal bite force and its association with signs and symptoms of craniomandibular disorders in young Finnish non-patients. AB - Maximal bite force values and subjective symptoms and clinical signs of craniomandibular disorders (CMD) were recorded for a non-patient sample of 129 young adults, 56 men and 73 women. The signs and symptoms of CMD were classified on the basis of Helkimo's clinical dysfunction index. The two genders reported equally frequently subjective symptoms of CMD, but women had significantly more severe clinical signs of CMD than did men. Mean maximal bite force values for men were 909 N (SD, 177) in the molar region and 382 N (SD, 133) in the incisal region and thus significantly higher than corresponding figures for women, 777 N (SD, 168) in the molar, 325 N (SD, 116) in the incisal region, suggesting that separate evaluation of the genders would be advisable in future studies involving bite force assessments. Neither subjective symptoms nor clinical signs of CMD correlated significantly with maximal bite force values. The bite force values measured were in line with theoretical calculations. PMID- 7484107 TI - Long-term follow-up of cross-arch fixed partial dentures in patients with advanced periodontal destruction. Evaluation of the supporting tissues. AB - The aim of the study was to evaluate the periodontal conditions in patients treated more than 10 years ago for advanced periodontal disease and rehabilitated with cross-arch fixed partial dentures (FPDs). Of 50 randomly selected patients, 34 subjects carrying 43 FPDs agreed to participate in a clinical and radiographic follow-up examination. The FPDs were divided into three groups on the basis of prosthesis design (end abutments, unilateral cantilever, and bilateral cantilevers) and amount of supporting tissues at the time of bridge installation. Seventy per cent of the FPDs were found to be unchanged, whereas the rest had been modified due to various complications leading to extraction of one or more of the abutment teeth. Six FPDs (14%) had been partially replaced by FPDs anchored to osseointegrated dental implants. A total of 21 (8%) of the original 274 abutment teeth had been extracted. Longitudinal changes in the amount of periodontal support were minimal over the average of 15 years of follow-up. FPD design or initial amount of supporting tissues was found not to have significant influence on longitudinal changes in periodontal conditions. It was concluded that combined periodontal and prosthodontic treatment of patients with advanced loss of periodontal support may provide a high rate of long-term successful outcome, provided proper adequate periodontal and prosthetic treatment and maintenance care are given. PMID- 7484110 TI - Effects of orthodontic magnets on cutaneous epithelial thickness and tibial bone growth in rats. AB - The use of permanent magnets in orthodontic treatment has increased significantly over the past years, triggered by the introduction of rare earth magnetic alloys. Concerns about possible side effects have been expressed relating to their use in orthodontic treatment. In previous experimental studies evidence of a reduction in bone formation and epithelial turnover close to rare earth magnets has been presented. The aim of the present study was twofold: to confirm earlier results and to study whether the observed changes are reversible. One hind leg in each of 45 rats was fitted with aluminum rings with Co5Sm magnets, whereas the other hind leg was fitted with sham rings. After 8 weeks 25 rats were killed, and the rings in the remaining rats were removed. The latter group was killed after another 11 weeks. The epithelial and bone reactions were evaluated histomorphometrically. Previous results showing thinner epithelium and retarded rate of bone formation close to Co5Sm magnets were confirmed. These effects were also shown to be largely reversible. It was concluded on the basis of this and the magnitude of the effects that any local side effects from orthodontic rare earth magnets appear negligible when extrapolated to the clinical situation. PMID- 7484111 TI - Corrected sagittal tomography of the temporomandibular joint. Influence of errors in film and patient positioning on linear and angular measurements. AB - The aim of this study was to assess the variation in film and patient positioning in horizontally corrected sagittal tomography of the temporomandibular joint (TMJ). The influence of this variation on linear and angular measurements of some anatomic structures of the TMJ was also studied. There was no significant variation in film positioning in the tomograph, using a multi-film cassette. The variation in positioning the patient in the tomograph was significant when four dental assistants were asked to place the Frankfort plane parallel to the horizontal plane. The measurements with the film in a straight position were compared with the measurements with the film angulated, to simulate the mean variation in film and patient positioning. Linear and angular measurements of anatomic structures were performed in arthrotomograms of 58 joints, representing joints with superior disk position and joints with anterior disk position with and without reduction. There was a difference between the two measurements for four different distances and one angle independent of diagnosis. No differences could be found between patients belonging to the different diagnostic groups. The results indicate that variation in patient positioning influences linear and angular measurements of anatomic structures in TMJ arthrotomograms. PMID- 7484112 TI - Photoelastic experiments on facings laminated to teeth. AB - The influence of various cervical designs of laminated facings was investigated. A variation in the attachment between tooth and facing was also investigated, to examine the influence of partly bad adherence of the resin. The facings were incisally loaded at three different angulations. The study was made in a polariscope, using an Na light source and quarter wave plates. Lack of adhesion of the laminate incisally or cervically created a higher fringe order than when the middle part suffered from insufficient bonding. The shape of the cervical part had only a minor effect on the stress distribution in the facing. PMID- 7484113 TI - Late results of the Souter-Strathclyde total elbow prosthesis in rheumatoid arthritis. 6/19 implants loose after 5 years. AB - We performed total elbow replacement, using the Souter-Strathclyde prosthesis in 19 elbows of 18 patients with rheumatoid arthritis and followed the patients for 5 (1-11) years. Pain relief was achieved initially in all patients. The average range of flexion-extension was increased by 12 degrees and pronation-supination by 40 degrees. There were no infections. 1 patient sustained an intraoperative fracture of the medial epicondyle, 3 patients developed neuropathies and 1 patient had an immediate postoperative dislocation of the joint. At follow-up, 6 prostheses had radiographic loosening, with sagittal titling and migration of 4 humeral components. 2 patients had clinical symptoms of loosening. PMID- 7484114 TI - Epidemiology of scapular fractures. Incidence and classification of 338 fractures. AB - We have studied scapular fractures, with special reference to intraarticular glenoid fractures, during a 10-year period in 2 Swedish counties. There were 338 scapular fractures in 322 patients. The annual incidence was 10/10(5) inhabitants, of which 30 percent affected the glenoid cavity. Out of 100 intraarticular glenoid fractures, 55 occurred in men and 45 in women. The mean age of the women at the time of fracture (64 years) was significantly higher than the mean age of men (49 years). The most common intraarticular glenoid fracture type was the anterior chip fragment fracture which, in about two-thirds of the cases, was associated with shoulder dislocation. PMID- 7484115 TI - Shoulder instability. Assessment of anterior-posterior translation with a knee laxity tester. AB - We developed a simple method to measure the anterior-posterior (AP) translation of the shoulder joint: by positioning the Donjoy Knee Laxity Tester horizontally over the shoulder girdle, the AP translation was measured after applying a standardized sagittal force. Measurement of AP-translation during two tests on the same shoulder was reproducible with an intraclass correlation coefficient (ICC) of 0.996. In normal subjects the mean side difference was 0.6 mm, corresponding to a p-value of 1.00. In patients with unilateral instability AP translation was significantly larger on the injured shoulder. Furthermore, the intraindividual differences in AP-translation were significantly larger in patients with unilateral instability than in normal subjects. Individuals with multidirectional instability had significantly higher values than any of the other groups. PMID- 7484116 TI - Surgery for nonunion of the lateral humeral condyle in children. 6 cases followed for 1-9 years. AB - Nonunion of the lateral humeral condyle in 6 children was treated by open reduction, internal fixation and bone grafting. All cases united. All patients had relief of pain. There were no complications. PMID- 7484118 TI - Recurrent kyphosis after posterior stabilization of thoracolumbar fractures. 24 cases treated with a Dick internal fixator followed for 1.5-4 years. AB - 24 patients with Th12-L1 fractures treated with a Dick internal fixator were analyzed to assess predictors of poor outcome. 4 patients had fixation without bone transplantation, 20 patients had a posterior fusion, and 12 of them had additional transpedicular spongioplasty. There were fractures of the transpedicular screws in 4 and screw migration in 2 cases. The increase in the local kyphosis angle was greater than the increase in the anterior compression angle and this did not correlate with spongioplasty or fusion. Fixation failure was in all cases related to a disproportionate increase in the local kyphosis angle. There was no difference between the patients with transpedicular spongioplasty and posterior fusion and the other patients with respect to results and complications. Bony collapse was not the major cause of failure and consequently there was no measureable contribution of transpedicular spongioplasty. We found that the Dick internal fixator for unstable fractures was associated with a higher complication rate than earlier reported. PMID- 7484119 TI - Electrical muscle stimulation on the spine. Three-dimensional effects in rabbits. AB - We investigated the 3-dimensional effect of electrostimulation of the latissimus dorsi, the erector spinae and the intercostal muscles on spinal configuration in 16 New Zealand white rabbits. Electrostimulation on the right side of the spine resulted in a left convex, hypokyphotic curve and vertebral body rotation towards the convexity of the curve in all rabbits. The Cobb angle in the coronal plane increased with stimulation of each of the muscles examined. The kyphosis decreased with stimulation of the latissimus dorsi and the erector spinae. The vertebral rotation increased with stimulation of all muscles. Stimulation of the tested muscles resulted in the simultaneous occurrence of a 3-dimensional spinal deformity with the characteristics of idiopathic scoliosis. PMID- 7484117 TI - Lumbar spinal stenosis. A retrospective study of 163 cases in southern Sweden. AB - I assessed the incidence of lumbar spinal stenosis as well as the frequency of severe neurological symptoms and signs based on patients seen in 2 orthopedic departments. The annual incidence in Malmo, 1982-1986, was 59 and in the period 1987-1991, 47 per million inhabitants. In Vaxjo, 1987-1991, the annual incidence was 45 per million. Severe neurological symptoms were few. Reduced EHL power and peroneal paresis were the most prevalent signs and were found in 13 and 12 percent, respectively, of all 163 patients. 1 patient had bladder dysfunction, 1 had impotence and 2 had a cauda equina syndrome. PMID- 7484120 TI - Radiographic loosening of cementless threaded acetabular cups. No additional diagnostic value of arthrography in 30 patients. AB - We compared arthrography with plain radiographs in 30 consecutive patients having a clinical diagnosis of loosening of a smooth-threaded acetabular prosthesis (Mecron, Berlin). Leakage of contrast at the interface between the ring and the bone on the medial side of the prosthesis was seen in 21 patients. Loosening of the cup was also visible on the plane radiographs and loosening was confirmed in all these patients at revision surgery. No false positive arthrographies were seen. In one patient, the arthrography was false negative because of a technical failure. We conclude that no additional information was obtained by arthrography. PMID- 7484122 TI - Stem loosening more common with ITH than Lubinus prosthesis. A 5-year clinical and radiographic follow-up of 142 patients. AB - Out of a consecutive series of 169 cemented total hip arthroplasties, 77 Lubinus SP 2 and 65 ITH prostheses, were followed-up after 5 years with clinical and radiographic assessments. Radiolucencies were commoner around the ITH titanium stem than around the Lubinus cobalt-chrome stem. 6 of the ITH stems were considered as definitely loose, as compared to none of the Lubinus stems. Hitherto, 5 patients in the ITH group have had revisions and 2 need revision because of loosening. The poorer performance of the cemented ITH stem may be due to an unfavorable combination of design and material properties. PMID- 7484121 TI - Accuracy of migration analysis in hip arthroplasty. Digitized and conventional radiography, compared to radiostereometry in 51 patients. AB - We assessed the accuracy of migration measurements on conventional and digitized radiographs of total hip arthroplasties by comparing the results with radiostereometry (RSA). 4 stem and 3 acetabular designs were studied. 2 of the stem designs and 1 cup design were uncemented. 180 manual and 202 digitized measurements were done on 120 conventional radiographic examinations. The readings on digitized radiographs did not differ from the manual measurements on the same radiographs. A comparison of the measurements from plain radiographs and with RSA of the femoral stems revealed an accuracy (absolute mean + 2 SD) of between 3.9 and 12.3 mm, depending on the choice of landmarks. The greatest accuracy was obtained by using tantalum markers inserted into the greater or the lesser trochanter and the shoulder of the stem. The most medial point of the lesser trochanter proved to be the best bony landmark. Measurements, including both the center of the femoral head and the greater trochanter, were associated with poor accuracy. The accuracy as regards horizontal cup migration varied from 4.4 to 6.5 mm and the accuracy as regards vertical migration varied between 4.4 and 6.3 mm. The intraobserver error (2 SD) ranged from 1.6 mm to 5.6 mm, The corresponding figures for the inter-observer error were 2.6 mm and 6.6 mm, respectively. One of the cemented cup designs was associated with inferior accuracy. Stem design did not affect the accuracy. PMID- 7484123 TI - Bone mineral density around femoral stems. DXA measurements in 22 porous-coated implants after 5 years. AB - In this cross-sectional study the bone mineral density (BMD) close to proximal porous-coated femur stems was measured by DXA 4.5-6 years after the stem implantation and compared to the contralateral non-operated femur in 22 cases. Measuring areas were defined by Gruen's method. The average precision error varied according to the zone assessed and ranged from 2.2 to 4.9 percent. The median BMD values of the operated femur were in 5 of 7 areas lower than those of the contralateral side. The largest differences were noted in the calcar region and the greater trochanteric region (21 and 20 percent, respectively). No correlation was found between the femoral BMD differences and the stem diameter. PMID- 7484124 TI - Stem fracture with the Exeter prosthesis. 3 of 27 hips followed for 10 years. AB - We report the results of a 9-11-year clinical and radiographic follow-up of 27 Exeter prostheses in which cement pressurizing technique was employed. 3 stems fractured and 3 other stems and 1 cup underwent aseptic loosening. The remaining prostheses had satisfactory clinical and radiographic results. Stem subsidence was seen only as part of a loosening process. We suspect that the pronounced taper design is responsible for the poor results. While the slender and weak distal end is fixed in a thick cement mantle, the wide proximal part allows only a thin cement layer, easily subjected to mechanical disintegration. Proximal debonding increases both the stress on the distal part of the stem and the distal bone-cement interface shear stress. Hence, we believe that the same process underlies both the loosenings and the stem fractures. The presently employed Exeter stem is manufactured from the stronger Orthinox steel, which may diminish the risk for fracture, but it has retained the extreme taper design. It is not likely that a polished surface or improved cementing will prevent stem fracture. Therefore, one should still be concerned about late fractures of the Exeter stem. PMID- 7484126 TI - Laparoscopic extraction of a displaced LIH pin. A case report. PMID- 7484125 TI - Hydatid disease of the pelvis and the femur. A case report. AB - We report a case of left-sided extended pelvic infestation of hydatid disease. The destruction of the hip joint, os pubis, larger parts of os ileum and the femoral head made pelvic resection and prosthetic replacement necessary. Recurrence of a hydatid cyst 5 years later in the left groin originating in the femur was treated with a second femoral resection and total hip replacement. Because of instability, the pelvic replacement also had to be changed to a custom made CAD replacement. This surgical procedure was combined with chemotherapy by mebendazole. The need for radical resection in osseal hydatid disease is discussed. PMID- 7484127 TI - A new osteotome for performing chevron trochanteric osteotomy. AB - We report on the use of a new modified osteotome for performing chevron trochanteric osteotomy during hip arthroplasty. It is easy to use and the apex of the chevron is directed distally, thus producing a large segment of osteotomized trochanter, while avoiding the problems of fragmentation and splintering. At the end of the procedure, the trochanter is easily seated back within its concave bed, automatically achieving anatomical reduction and allowing easier fixation using the cruciate wiring technique. In a series of 77 primary hip arthroplasties, bony trochanteric union was achieved within 6 months in all patients. PMID- 7484128 TI - Infected tibial nonunion. Good results after open cancellous bone grafting in 37 cases. AB - We treated 37 infected tibial shaft nonunions by debridement followed by open autogenous cancellous bone grafting in a 2-stage procedure. Additional surgery was done in 21 fractures including second debridement before bone grafting and/or a second limited bone grafting and/or a split-thickness skin grafting. All fractures healed after an average of 11 (8-16) months. During 2 years follow-up there were no recurrences of the infection. Two cases of early refracture occurred, both healed following new bone grafting. PMID- 7484129 TI - Long-term outcome after ruptured lateral ankle ligaments. A prospective study of three different treatments in 79 patients with 11-year follow-up. AB - In a prospective study of 79 patients with arthrographically verified acute ruptured lateral ankle ligaments we have evaluated the long-term results of 3 different treatments: operation and walking-cast for 5 weeks, walking-cast alone, and elastic bandage. The follow-up period was 11 (9-13) years. Residual disabilities and late complications, such as instability, pain on activity, and the number of ligament reconstructions and talocrural arthroses were equally low in all 3 groups. We conclude that nonoperative treatment seems adequate. PMID- 7484130 TI - Injury location affects ligament healing. A morphologic and mechanical study of the healing rabbit medial collateral ligament. AB - Based on the heterogeneity of the rabbit medial collateral ligament (MCL) along its length, we tested the hypothesis that injury location would affect its healing response. The right MCL of 80 skeletally mature New Zealand white rabbits was sectioned adjacent to bone at the femoral end (40 rabbits) or the tibial end (40 rabbits) and reapposed with sutures. Animals were killed after 3, 6, 14, or 40 weeks of healing to examine wounds histologically (2 rabbits per healing interval) and mechanically (8 rabbits per healing interval). Results of the mechanical tests were compared to midsubstance MCL repairs (24 rabbits) and to uninjured normal MCLs (20 rabbits). The morphology of the near-insertion repairs was characterized by abnormal callus-like formation and patchy bone resorption, particularly at the tibial insertion. Mechanically, insertional injuries remodeled towards normal MCL low-load, viscoelastic and failure properties more slowly than midsubstance injuries at the early healing intervals. After 40 weeks of healing, few injury-specific differences persisted. All injured ligaments had ultimate strengths 15-35 percent short of normal at 40 weeks and the femorally injured ligaments were weaker than normal at this time. These results suggest that rabbit MCLs, injured near either end, heal more slowly than those injured in their midsubstance and develop abnormal insertion morphology. PMID- 7484131 TI - Bone formation in rabbit cancellous bone defects filled with bioactive glass granules. AB - We examined new bone formation after filling cancellous bone defects with bioactive glass (BG) in granular form. Cylindrical defects in the trochanter area of 18 rabbit femora were filled with BG granules (diameter 600-830 microns) and compared with similar defects filled with morcellized autogenous bone. New bone formation and surface reaction of BG particles were evaluated by light microscopy, histomorphometry, and scanning electron microscopy. The chemical profile at the bone- material interface was studied by energy dispersive x-ray analysis (EDXA). In the BG group, 41, 32, and 38 percent of the defects were filled with new bone after 3, 6, and 12 weeks, respectively. The corresponding figures for the autogenous bone group were 36, 29, and 34 percent. The thickness of the reaction layer on the glass surface increased from 82 to 163 microns during the observation periods. An intimate contact without intervening soft tissue between new bone lamellae and BG granules was a constant finding. EDXA showed a chemical continuum between the granules and the new bone. No adverse reactions related to BG were observed. BG is a promising material for filling cancellous bone defects. PMID- 7484132 TI - Periosteal response to transient ischemia. Histological studies on the rat tibia. AB - Complete arrest of blood flow was induced for 4.5 hours in the left hindlimb of Wistar rats by a tourniquet applied proximally to the thigh. Histologically the periost of the tibial bone after 3 days showed marked hypertrophy and hyperplasia of the periosteal cells with an osteogen differentiation. This response is similar to the initial formation of external callus during fracture healing. Transient ischemia may be an important factor in initiating fracture healing. PMID- 7484133 TI - Oxygen-free radicals impair fracture healing in rats. AB - We studied the effect of oxygen-free radicals on fracture healing. 30 male rats were divided into 2 groups: 15 rats were given saline 5 mL/kg i.p. (control group) and 15 were given zymosan 100 mg/kg i.p. to induce oxygen-free radicals through stimulation of NADPH oxidase in polymorphonuclear leucocytes. 1 hour later, the right forelimbs of the rats were broken by light manual compression. These treatments were given once a day until the fifth post-fracture day. All rats were killed on day 22, and histological sections of the radius and ulna were examined without knowledge of the treatment given. The administration of zymosan impaired the fracture healing and therefore oxygen-free radicals appear to play an important role in fracture healing. PMID- 7484134 TI - A screw for cervical hip fractures designed to minimize migration. PMID- 7484135 TI - Callus distraction in Ollier's disease. PMID- 7484136 TI - Stress fracture of the patella. PMID- 7484137 TI - Forearm fracture due to the release of an automobile air bag. PMID- 7484138 TI - Massive exposition to titanium, but without sensitization. PMID- 7484139 TI - Management of nasal polyps: is objective assessment possible? PMID- 7484140 TI - The role of endoscopy in the management of paranasal sinus tumours. AB - Only recently management of benign as well as malignant nasal and paranasal sinus tumours by endoscopy, including endoscopic removal, has been introduced. The most important contribution of the endoscope in the management of these tumours is diagnostic, primarily as well as during follow-up. Although, in general, endoscopic sinus surgery has many advantages over classical external surgery, disadvantages are present when tumours are concerned. Some agreement already exists on the role and limitations of endoscopic sinus surgery in benign tumours, but in malignant processes usually more mutilating procedures are needed and only in selected cases might endoscopic removal be used. PMID- 7484141 TI - The kinetics of the cytologic and rhinomanometric changes in the nose after challenge with allergen. AB - The objective of these experiments was to study the dynamics of the cytologic and rhinomanometric nasal reaction after allergen challenge. For this purpose a micro suction system was developed to repetitively and atraumatically collect nasal secretions. Outside of the pollen season, four patients allergic to grass pollen were challenged with phosphate buffered saline (PBS) and, two weeks later, with grass pollen. Six control subjects were challenged with PBS only. Repetitive nasal secretion samplings over three days and rhinomanometry (PAR) were performed. Analysis of samplings taken before allergen challenge proved the atraumatic character of the micro-suction and provocation methods. Coupling the obtained dynamic data of nasal cytology and PAR, the following observations could be made: 1) Higher number of eosinophils in patients in the absence of antigen, compared to controls, suggesting a constitutional characteristic of atopy. 2) A close temporal association between cytological and clinical events occurring during the late phase of the allergic reaction. Nasal obstruction seems to be correlated with the influx of eosinophils rather than with their absolute concentration. PMID- 7484142 TI - Phenomenology, pathogenesis, diagnosis and treatment of aspirin-sensitive rhinosinusitis. AB - Aspirin-sensitive rhinosinusitis is a non-allergic, non-infectious perennial eosinophilic rhinitis starting in middle age and rarely seen in children. It may also been seen in atopic patients who have developed a mixed type rhinitis with recurrent airway infections. There is an intolerance to aspirin and most other NSAID. An intolerance to tartrazine, food additives, alcohol, narcotics and local anaesthetics can follow. Most aspirin-sensitive patients develop nasal polyps. Untreated, it can lead to asthma. The frequency of aspirin intolerance is 6.18% in patients with perennial rhinitis and 14.68% in patients with nasal polyps. Immunologic studies of the blood and the nasal polyps show a hyperreactive immune system with an activation of the eosinophil granulocytes due to a TH1-lymphocyte activation. In atopic subjects with a mixed type rhinitis, we found a TH2- and B lymphocyte-activation as well. Inhibition of eosinophil apoptosis might be a second remarkable change in the immune system of aspirin-sensitive patients. A key pathogenic event for aspirin sensitivity is the change of the leukotriene pathway for arachidonic acid metabolism releasing high amounts of leukotrienes LTC4, LTD4 and LTE4, effective chemoattractants and activators of inflammatory cells. For the diagnosis of aspirin intolerance, nasal, bronchial and oral challenge are available. The sensitivity of nasal challenge with lysine-aspirin for the diagnosis of aspirin-sensitive rhinitis is 0.93, the specificity 0.97. It is the safest test in aspirin-sensitive asthmatics causing bronchial side effects only in 0.45%. Therapy of aspirin-sensitive rhinosinusitis includes avoidance of aspirin and NSAID. A general down regulation of the immune response with glucocorticosteroids is an effective means. We prefer a maintenance dose of budesonid 400 micrograms a day. Systemic steroids for a reversibility test or in exacerbation due to viral infection are given in a dose of 50 mg a day for one week. If steroids don't work well enough, we combine them with aspirin desensitizations at a maintenance dose of 500 mg a day. Gastrointestinal side effects occur in 20% of the patients with a dose of 500 mg aspirin a day, in 46% with a mean dose of 1300 mg a day. The combined treatment of topical nasal steroids and aspirin-desensitization is effective in 65% of the patients with improvement in the symptoms of hyper-secretion, irritation and blockage, while 73% show improvement of polyps, hyposmia and anosmia. Endonasal endoscopic surgery of the ethmoids, turbinectoms and septoplasty should be done if necessary, especially in cases where conservative treatment fails. After surgery a further antiinflammatory treatment is absolutely necessary otherwise polyps reoccur in 90% of the cases after weeks or months. Unfortunately there is so far no curative treatment. New drugs like cytokine or leukotriene receptor antagonists give hope for better results in treatment of aspirin intolerance in the future. PMID- 7484144 TI - Complete sphenoethmoidectomy and computer-assisted surgery. AB - Many surgical procedures in the field of ENT take place in close proximity to vital structures like the orbit, the skull base, the internal carotid artery etc. In our clinic we have decided to study two different computer-assisted navigation systems to reduce the risk of trauma to these structures during endonasal endoscopic procedures. Such systems should be able to correlate the position of the surgical instrument, ideally in the submillimeter range, to CT- or MR-images. The ARTMA Virtual Patient finds the position of the instrument by permanently measuring magnetic fields. The ISG-system uses a mechanical arm to localize the probe in the patient. Using Computer-Assisted-Surgery (CAS) does not significantly extend the time needed for surgery. We found that it prevents the surgeon from inadvertently injuring structures. Other groups have shown (1) that CAS-systems minimize the risk of complications of surgical procedures in the frontobasis arising from mishaps in the usage of the endoscope or the surgical tool. If properly used, CAS-systems can be a very helpful tool in the hand of an experienced surgeon but will never replace his expertise and knowledge. PMID- 7484143 TI - Nosocomial pansinusitis in orotracheally intubated critically ill patients. AB - Nosocomial pansinusitis (N P) is most often described in nasotracheally intubated patients with craniocerebral or facial trauma. We retrospectively reviewed its occurrence and complications in the course of prolonged mechanical ventilation in orotracheally intubated patients without maxillofacial or cranial injuries. N P was deemed to be present when (1) CT scan showed opacification and/or air-fluid levels in the maxillary, ethmoid and sphenoid sinuses and (2) aspiration of both maxillary sinuses yielded pus, cultures of which revealed a high concentration of micro organisms. Nosocomial pneumonia and bacteremia were considered related to the N P if the organisms found in the sinus were identical to those recovered from the blood and/or the bronchi. During an 18-month study period, 38 cases of sinusitis were diagnosed. N P was present in 13 patients: 18 organisms (12 Gram negative, 5 Gram-positive and 1 Candida albicans) were isolated. Pneumonia occurred in 8 patients, 6 with multi-resistant Pseudominas aeruginosa and 2 with methicillin resistant Staphylococcus aureus. Pseudomonas aeruginosa was isolated from the blood, lung and sinus in two patients. This study demonstrates that N P is relatively frequent in patients requiring long-term mechanical ventilation, even in the absence of predisposing factors. N P in these patients is mostly monomicrobial with multi-resistant Pseudomonas aeruginosa and Staphylococcus aureus as the main causative agents. PMID- 7484145 TI - Results of the endonasal surgical closure of nasoseptal perforations. AB - Eleven patients with nasal septal perforations were surgically treated within a period of seven years. Former septal surgery was the major cause of nasoseptal perforations. The technique most frequently used was a Cottle septoplasty with closure of the perforation by bipedicled flaps and interposition of autogenous cartilage and bone. The authors were inspired by Younger and Blokmanis's model and they used the following parameters to evaluate the results: location, size, symptoms pre- and post-operatively and also rhinoscopia pre- and post operatively. In nearly all cases the perforation was closed with a decrease of the symptoms. PMID- 7484146 TI - Early management of the paralyzed upper eyelid using a gold implant. AB - One main goal in the management of a peripheral facial paralysis is to provide the patient with adequate protection to the cornea in order to avoid keratitis. Medical treatment, (ophthalmic drops, ointments, taping, etc.), often fails and surgery has to be considered. The various methods hitherto described, e.g. tarsorraphy, plastic strips, magnetic implants and wire springs, are associated with drawbacks such as infection, poor cosmetic outcome or need for a revision procedure. Between 1990 and 1992, 12 patients, suffering from an upper eyelid paralysis, as a consequence of a peripheral impairment of the homolateral facial nerve function, were operated on in our Department with the implant of a gold weight within the upper eyelid itself. This procedure yielded, in all cases, an excellent lid closure, corneal protection as well as a good cosmesis. The surgical technique is simple, if performed under local anesthesia, and the patient leaves the hospital on the same day of surgery. Should the natural eye function return, the implant can be easily removed. PMID- 7484147 TI - Obstructive sleep apnea syndrome in children: an overview. AB - All children are at risk for the development of an obstructive sleep apnea syndrome (OSAS) since adenotonsillar hypertrophy is widespread and the most common cause in children. The clinical picture, etiology and diagnostic approach of OSAS in children is discussed. Attention is given to the indications for polysomnography and the differences between adult and pediatric OSAS. An overview is presented of the different treatment modalities and the possible link between sudden infant death syndrome, pediatric OSAS and adult OSAS. PMID- 7484149 TI - The accuracy of tympanometry assessed by its sensitivity and specificity. AB - Tympanometry has become a widely accepted diagnostic tool for the diagnosis of otitis media with effusion (OME). The present study was conducted to assess the accuracy of the technique by calculating its sensitivity and specificity. Forty children with chronic OME and recurrent otitis media were studied. Immediate preoperative tympanograms were compared with operative findings of tympanocentesis, and sensitivity and specificity data were calculated. Approximately one third of the B type tympanograms (effusion) changed to A (effusion free) or C type (possible effusion) during the waiting period for the operation, showing resorption of fluid from the middle ear cavity. Sensitivity score was 95% bearing out the C type tympanograms. When C type tympanograms were included into the calculation, sensitivity dropped to 73%, showing liquid frequently present in cases of C type tympanograms. Tympanometry scored as more reliable in the presence of adenoid hypertrophy, when sensitivity was 83% and specificity was 90%. The absence of adenoid hypertrophy led to a drop of sensitivity to 58% and to a drop of specificity to 71%. These results may indicate that inhalant induction of anaesthesia has less effect on the middle ear in the presence of adenoidal hypertrophy and concomitant eustachian tubal dysfunction. Tympanometry proved to be a reliable diagnostic tool for the diagnosis of otitis media with effusion. Long preoperative waiting periods and the absence of adenoid hypertrophy could be responsible for the differences between preoperative tympanograms and the middle ear status observed at tympanocentesis. PMID- 7484150 TI - Experimental evidence of ototoxicity of ear drops. A review of the literature. AB - Evidence of ototoxicity as derived from laboratory experiments on animal models and from a few prospective controlled studies in man is reviewed. Data are gathered for the different components and antibiotics contained in ototopical preparations. An attempt is made to answer the question whether these laboratory findings are valid in the clinical situation in man. PMID- 7484148 TI - The tympanic wink (le clin d'oeil tympanique). AB - The "tympanic wink" illustrates a beating brillance born on the antero-inferior quadrant of the promontory and appearing on the tympanic membrane as a false beating light triangle. This sign may be an aid in discovering a minor vascular abnormality of the middle ear: hypertrophy of carotico-tympanic or inferior tympanic arteries, dehiscence of the ascending part of the internal carotid canal or the possibility of a persistent stapedial artery. PMID- 7484152 TI - [Morphologic and ultrastructural changes of soft palate in patients who underwent palatopharyngoplasty]. AB - In order to clarify pathogenesis of Obstructive Sleep Apnea Syndrome (OSAS) in patients with anatomic abnormalities of upper airways, we studied soft palate and uvula of OSAS patients by means of histological and ultrastructural techniques. 38 OSAS patients, severe and moderately severe form, underwent modified Palatopharingoplasty. We evaluated only 16 OSAS patients' soft palate and uvula and observed histological changes in all of them: submucosal edema and minor salivary glandes ipertrophy and iperplasy are present. On the contrary, ultramicroscope showed normal muscle fibers. Strie Z alterations and sarcomeres disorganization, although present, have no statistical value. Very probably, ronflement and apneas determined the observed alterations through trauma on pharingeal wall. Therefore, they are not OSAS primary cause. PMID- 7484151 TI - [Brain stem auditory evoked potentials in obstructive sleep apnea syndrome]. AB - To evaluate a possible brainstem role in pathogenesis of obstructive sleep apnea syndrome, a study on brainstem auditory evoked potentials (BAEP), has been conducted. 15 OSAS patients, medium and severe form, with organic pathologies of the upper respiratory tract have been evaluated. 15 normal subjects were compared as control. All the patients were subjected to audiometry, including total liminal audiometry, timpanometry, acustic reflex, and BAEP study. BAEP evoked with trains of cliks at 11 and 51 periods/sec., showed morphological alterations and a longer central conductance of time interval (I-V interval) in only four patients. BAEP alterations noted in the OSAS-affected patients are neither constant nor specific. Therefore, the observed BAEP alterations might be due to apneas, as a consequence of the chronic hypoxic- hypercapnic status occurring in the brain-stem. PMID- 7484153 TI - [The surgical treatment of obstructive sleep apnea syndrome]. AB - Studies concerning the anatomical abnormalities of upper airways in patients affected by Obstructive Sleep Apnea Syndrome (OSAS) allowed the corrective surgical treatments in this syndrome. To provide an improvement or a definitive recovery of OSAS, various surgical treatments, i.e. functional nasal surgery, palatal surgery [Partial resection of palate (RPP), Uvulopalatopharingoplasty (UPPP), Palatopharingoplasty (PPP), modified Palatopharingoplasty (modified PPP)], mandibular and base tongue surgery, have been performed in order to obtain an enlargement of upper airways. It has been reported that in RPP, UPPP, PPP and modified PPP partial or total uvula and soft palate resection is conduct; in our research we performed modified PPP. 44 OSAS patients, severe or moderately severe form (polisomnographic diagnosis), underwent surgical treatment: 5 patients underwent functional nasal surgery; 9 patients underwent modified PPP; 29 patients underwent nasal and palatal surgery; only 1 patient underwent base tongue surgery. 32 patients underwent polisomnographic records after surgery (two months later) and we evaluated Apnea Index (I.A.) and a SaO2 low (nadir) with statistic tests. The results have been positive. Therefore, we have now 7 normal, 2 mild form, 7 moderate form, 3 moderately severe form and 13 severe form OSAS patients. If, on the one hand, all these surgical treatments can be considered a good way of therapy, on the other, only tracheostomy represents today the unfailing surgical therapy. PMID- 7484154 TI - [The cardiovascular reactions mediated by TPA and tamoxifen in spinal cord of conscious rats]. AB - Intrathecal administration (ith) of phorbol 12-myristate (TPA), an activator of protein kinase C, (31.24, 26.5, 125 and 250 ng/10 micrograms) to conscious rats produced a marked dose-dependent pressor effect without significant change in heart rate (HR). Intrathecal administration of tamoxifen (3.125, 6.5 and 100 micrograms/10 microliters), one of the inhibitors of protein kinase C, produced a marked dose-dependent hypotensive effect. Intrathecal injection of exogenous DBcAMP (12.5, 25, 50 and 100 micrograms/10 microliters) also increased the mean arterial blood pressure (mABP) and markedly lowered HR. It was further observed that, based on the effect of DBcAMP (25 micrograms/10 microliters), the pressor effect of four doses of TPA can be augmented. The results indicate that TPA in spinal cord may exert hypertensive effect via activation of PKC and tamoxifen may exert a hypotensive effect via inhibition of PKC. In addition, the pressor effect of TPA could be augmented by preadministration of DBcAMP. PMID- 7484155 TI - [Effects of neferine and its combination with taurine on platelet aggregation and experimental thrombosis in rats]. AB - The effects of neferine (Nef), taurine (Tau) and Nef + Tau on platelet aggregation and generation of thrombosis were investigated in rats. Using turbidimetry Nef and Tau were found to inhibit platelet aggregation induced by adenosine diphosphate (ADP), collagen or thrombin. Both drugs reduced the wet weight of experimental thrombosis. But the drugs showed no obvious effect on fibrinogen content and euglobulin lysis time. Thromboxane A2 (TXA2) production induced by ADP and prostacyclin (PGI2) were studied by enzyme immunoassay in rats. TXA2 generation in platelet rich plasma from rats treated with Nef and Nef + Tau obviously decreased. While the plasma PGI2 was not affected. Nef + Tau iv in 30 min significantly decreased the level of total cholesterin in serum. The results suggest that Nef has inhibitory effect on platelet aggregation and thrombosis formation. The combination of both drugs was found to be more potent than either one alone. The mechanism may be related to its reduction of TXA2 formation. PMID- 7484156 TI - [Cytocidal effect of HH07A, a derivative of hainanensine, on L1210 cells in vitro]. AB - The L1210 cells rapidly ceased to proliferate and its mitotic index decreased markedly after being exposed to HH07A 2 micrograms . ml-1 during exponential growth phase. However, 34.6% of the cells were still able to form colonies in soft agar if HH07A was removed after 24 h of incubation (the colony-forming efficiency for control cells was 63.7%) and the cell viability remained at about 94%. The DNA contents in L1210 cells were measured with a flow-cytometer. The results showed that the cell-cycle was blocked at the S phase and the number of cells in G2 + M phase decreased significantly. The time response course for L1210 cells indicated that the inhibitory effect of HH07A on L1210 clonogenic cells was slightly time dependent. PMID- 7484157 TI - [The pharmacokinetics and pharmacodynamics of cefotaxime in experimental diabetic rats]. AB - The present paper describes the pharmacokinetics and pharmacodynamics of CTX in both normal control rats and early alloxan-diabetic rats in vivo. The plasma concentrations of CTX were determined with HPLC. The main pharmacokinetic parameters of the two groups were calculated with computer. Compared with the control group, the C(m) and AUC in the diabetic group decreased significantly. Most of the free drug was distributed to tissue fluid because the protein binding of CTX decreased. But, the T1/2ka and T1/2ke of the two groups were not significantly different. It was suggested that the absorption and elimination rate of CTX were not affected in early diabetic rats. The PD50 of CTX against S. Pneumoniae infection was not different significantly between the two groups. This implies that the therapeutic effect of CTX was enhanced as the plasma free concentration could be increased in diabetic rats. PMID- 7484158 TI - [Studies on CDDP-albumin microspheres for hepatic arterial chemoembolization]. AB - An optimum procedure was established by orthogonal test for preparing cis-platin albumin microspheres (CDDP-AMS) with emulsion-heating stabilization method. The factors which affect particle size and release rate in vitro were studied. The particle size focusing on 58.8-256 microns, the mean size was 148.46 microns, drug content was 51.16% (w/w). The dissolution profiles of the CDDP-AMS followed Higuchi kinetics. In rabbits the distribution and elimination half times of platinum were prolonged 3.36 times and 1.23 times vs injection group, respectively, after hepatic arterial chemoembolization with CDDP-AMS. The highest serum concentration of platinum is 30 percent of that of the injection group. The platinum concentration was increased in liver (P < 0.01) and decreased in kidney (P < 0.05) vs that of injection group. PMID- 7484159 TI - [Studies on lung targeting gelatin microspheres of mitoxantrone]. AB - Lung targeting gelatin microspheres of mitoxantrone (DHAQ) were prepared by a two step method. The diameter of 87.36% of the DHAQ gelatin microspheres (DHAQ-GMS) was in the range of 5.1 to 25.0 microns. Release of the drug from the DHAQ-GMS in vitro became much slower and its t1/2 was 4 times longer than that of pure DHAQ. The characteristic peak of heat absorption on the differential thermal analysis curve was at 133 degrees C and almost no change was observed after the DHAQ-GMS were stored for 3 months at 37 degrees C (relative humidity 75%). The distribution test in vivo in mice indicated that the lung targeting effect of the DHAQ-GMS was obvious and that the targeting efficiency of the lung compared to other organs and blood increased 3 to 35 times. Kinetic behavior of the drug in mouse lung could be described by one open compartment model, and the average residual time increased by 10 h. PMID- 7484160 TI - Study of the influence of the auxiliary substances on the surface tension of the aqueous solutions of some substituted phenylcarbamic acid derivatives. AB - In the course of the drug form development of potential drugs the influence of auxiliary substances, namely glycerol (GL), propylene glycol (PG) and sorbitol (SO), used in the form of 5, 10, 15 and 20% solution on the surface tension (gamma) of some phenylcarbamic acid aqueous solutions was studied. Since this influence depends not only on the kind and concentration of the auxiliary substance used but also on potential drugs themselves, attempt was made to find optimal additives for particular drugs. The obtained results show that as optimal additives may be used: SO for IR-1 and H + B, for XIX M it is either PG or GL, for XIII it is GL, and finally for XIX it is either PG or SO. PMID- 7484161 TI - [Role of human cytochrome P-450 enzymes in the metabolism of xenobiotics]. AB - Cytochrome P450 enzymes play essential role in metabolism of exogenous compounds. They are found mainly in endoplasmic reticulum of hepatocytes. Purification of different P450 isoforms and determination of their structures and catalytic activities allowed to investigate their participation in metabolism of xenobiotics. Numerous factors cause variations in the expression of these P450: the effect of genetic and environmental factors results in the production of series of enzymes catalyzing biotransformation of xenobiotics. Several genetic polymorphisms have been described: CYP2D6, CYP2C and 'Ah locus' polymorphism. Some P450 (CYP1A, CYP3A, CYP2E1) are inducible by xenobiotics. Additionally, P450 expression and inducibility vary depending on tissues. It is now possible to determine which isoform(s) is (are) responsible for the production of a metabolite and to predict the fate of a compound in vivo. PMID- 7484162 TI - [Polymer-containing eyedrops. IV. Microbiological study of eyedrops containing antibiotics]. AB - Authors studied microbiological activities of eye-drops containing neomycin sulphate, gentamicin-sulphate and chloramphenicol, respectively. Effects of viscous eye-drops on Bacillus subtilis were measured by comparing them with resistest disc in the function of the type and concentration of macromolecule, pH and electrolyte content of the solution. It was found that viscosity increasing solution does not have any significant effect on the microbiological activity of the eye-drop in the studied range of concentration. Effectivity of the solution does not change either during storage. PMID- 7484163 TI - [Interaction between Bordetella pertussis vaccine and chlorpromazine in mice]. AB - In acute toxicity experiments authors observed increased mortality on the first day following 75 mg/kg chlorpromazine (CPZ) treatment in mice pretreated with Bordetella pertussis vaccine (9 x 10(9) killed bacterium/mouse) compared to control animals treated with CPZ alone. Initially, the increased drug sensitivity observed after combined treatment was attributed to summation of the toxic effects. However, the cumulation of mortality did not cease on the following days; furthermore, an increase of bacterial translocation (translocation index of P-CPZ group: 4.5) was observed on days 6 and 7, i.e. when the lymphocytosis, splenic hypertrophy and shrinkage of thymus--changes consequent to the vaccination--were at their maximum levels. On the basis of all these and on literary data it is supposed that the early cumulation of deaths after combined treatment may be in connection with an interaction between the two agents and that the side-effects following vaccination of humans may be induced by undesirable pharmacological interactions. PMID- 7484164 TI - [Methods of thin layer chromatography in quality control (a comparison of the Hungarian and European pharmacopoeias)]. AB - The role of TLC has been enhanced in pharmacopoeial analyses relative to its position in the analytics generally. So it has became on outstanding method in both Hungarian and European Pharmacopoeias. More than 400 purity control prescriptions of the monographs have been evaluated, the limiting tests classified and interpreted by the authors pointing out the similarities and differences. The purpose of the paper was to contribute to the preparation of joining of Hungary to the European Pharmacopoeial Convention. PMID- 7484165 TI - [Synthesis of potential CCK antagonist quinazolone derivatives. II]. AB - An alternative route has been developed for the synthesis of [1,4]diazepino[3,4 b]quinazolones (2), a new ring system of heterocondensed quinazolones. 2,3 Bifunctional quinazolones (4) were synthesized by halogenation of 2-alkyl quinazolone-3-propionic acids, yield from the reaction of 2-alkyl-benzoxazone (3) with beta-alanine. The reaction of haloalkyl compounds (5,6) with N-nucleophiles produces [1,4]oxazepino-[3,4-b]quinazolones (8), a new heterocyclic ring system, and 3-[2'-hydroxi-ethyl-, or 3-[2'-chloro-ethyl-4' oxo(3'H)-quinazoline-3' yl]propionamides (7, 9) in consecutive reaction. The cyclisation of 2-aminoalkyl 3-propionates (13) resulted in the title compounds (14). Physical data and properties of the newly synthesized compounds such as IR, UV and NMR spectra were obtained for all compounds reported. PMID- 7484166 TI - Structure-function relationships in the Ca(2+)-ATPase of sarcoplasmic reticulum studied by use of the substrate analogue CrATP and site-directed mutagenesis. Comparison with the Na+,K(+)-ATPase. PMID- 7484167 TI - Transcranial Doppler ultrasound for cerebral perfusion. AB - By providing a non-invasive method for continuous display of mean flow velocity (Vmean) in the cerebral arteries, transcranial Doppler (TCD) ultrasound supplements evaluation of cerebral perfusion. Dynamic exercise increases middle cerebral artery (MCA) Vmean from approximately 55 to 65 cm s-1 dependent on work rate, and even more when corrected for changes of the arterial carbon dioxide tension. Evaluation of Vmean corresponds to that of cerebral blood flow as determined with the 133Xenon clearance technique, and reflects regional cortical regulation of the active muscles with important afferent nervous influence. Concomitant increases of mean arterial pressure (MAP) and heart rate is only of minor importance as illustrated during static exercise and post-exercise muscle ischaemia, where Vmean is not significantly elevated. During sustained head-up tilt, the Vmean remained unchanged at a MAP approximately 83 mmHg. Below this level, it decreased in parallel with MAP until MAP reached 50 mmHg. At an even lower MAP, Vmean seemed to approach a lower limit approximately 25 cm s-1, but at a diastolic pressure of 21 mmHg there was no flow in the MCA. Conversely, during post-exercise muscle ischemia, an increase in MAP to 140 mmHg did not influence Vmean. This is in contrast to patients operated for carotid artery stenosis and who develop ipsilateral headache. In these patients the ipsilateral MCA Vmean changed in parallel with MAP, and autoregulation was re-established only after one to two weeks. In patients with severe carotid stenosis and poor collateral circulation, the CO2-reactivity as expressed by Vmean was the lowest, and could be negative on the ipsilateral side. During carotid endarterectomy, a Vmean clamp/Vmean pre-clamp ratio below 0.6 identified patients with a cerebral blood flow below 20 ml 100 g-1 min-1. Furthermore, when the ratio was below 0.4 pathological electroenchephalographic changes developed. Thus, Vmean of the large basal cerebral arteries reflects cerebral perfusion with respect to regional flow distribution, autoregulatory response, and CO2-reactivity in normal man and patients with limited cerebral flow. PMID- 7484168 TI - Isokinetic hamstring/quadriceps strength ratio: influence from joint angular velocity, gravity correction and contraction mode. AB - This study investigated isokinetic peak- and angle-specific hamstring/quadriceps strength ratios (conventional H/Q ratio) obtained during concentric and eccentric muscle contraction and examined the influence of joint angular velocity and the effect of gravity correction on these ratios. Also, a 'functional' H/Q ratio was defined by calculating eccentric hamstring strength relative to concentric quadriceps strength (Hecc/Qcon' representative for knee extension) and calculating concentric hamstring strength relative to eccentric quadriceps strength (Hcon/Qecc' representative for knee flexion). The H/Q ratio was calculated based on isokinetic peak moment and 50 degree-moment (0 degree = full extension) obtained at joint angular velocities 30, 120 and 240 degrees s-1. Gravity corrected conventional H/Q ratio remained constant across speeds and contraction mode, ranging from 0.47 to 0.54 and from 0.49 to 0.56 based on peak and 50 degree moment, respectively. In contrast, non-corrected H/Q ratio increased during concentric contraction from 0.58 at 30 degrees s-1 to 0.74 at 240 degree s-1 (P < 0.01). For knee extension at 240 degrees s-1 an Hecc/Qcon of 1.05 (peak) and 0.89 (50 degrees) was observed while for flexion at 240 degrees s 1 an Hcon/Qecc of 0.27 (peak) and 0.28 (50 degrees) was observed. In conclusion, gravity correction had high influence on the change in H/Q ratio with variation in extension velocity. A potential 1:1 hamstring/quadriceps strength relationship was demonstrated for fast knee extension, indicating a significant functional capacity of the hamstring muscles for providing muscular stability at the knee joint in such situations. PMID- 7484169 TI - Modulation of vascular contractile responses to alpha 1- and alpha 2-adrenergic and neuropeptide Y receptor stimulation in rats with ischaemic heart failure. AB - In order to evaluate adaptational changes in vascular function in congestive heart failure (CHF), we studied the contractile responses of isolated arterial and venous blood vessels from rats suffering from CHF induced by coronary artery ligature, resulting in a myocardial infarction. The contractile responses of the basilar, femoral and renal arteries and of the iliac vein were examined in relation to adrenergic and neuropeptide Y (NPY) receptor function by the action of the alpha 1 agonist phenylephrine, the alpha 2 agonist clonidine and NPY. The contractile force was measured (in mN) and in % of K(+)-induced contraction as well as pD2 to each agonist. When stimulated by a 60 mM K(+)-buffer solution, the femoral and renal arteries from CHF rats responded with a stronger contraction (Emax; 9.4 +/- 0.6 and 9.8 +/- 0.6 mN) than the corresponding Sham vessels (Emax; 6.2 +/- 0.7 and 5.6 +/- 0.4 mN respectively, P < 0.001). On the contrary, the iliac vein of CHF responded less to K+ than the Sham iliac vein (Emax 2.5 +/- 0.2 and 3.7 +/- 0.5 mN, P < 0.01). The CHF iliac vein responded with a weaker contraction when stimulated with phenylephrine (Emax 1.9 +/- 0.4 mN) and showed a lower sensitivity (pD2 5.6 +/- 0.1) than the corresponding sham vessel (Emax 5.7 +/- 2.3 mN and pD2 6.3 +/- 0.5, P < 0.05). The CHF renal artery was less sensitive to clonidine (pD2 6.4 +/- 0.6) than the Sham renal artery (pD2 7.2 +/- 0.1, P < 0.05). The results indicate differences between CHF and Sham vessel segments according to both contractile capacity induced by K(+)-depolarization and to agonist induced contractile capacity and sensitivity. The differences are not of general nature but vary according to the vascular bed examined. PMID- 7484170 TI - Rat skeletal muscle metabolism in experimental heart failure: effects of physical training. AB - Skeletal muscle metabolic abnormalities exist in chronic heart failure. The influence of physical training on muscle metabolism after myocardial infarction was studied in a rat model. 31P magnetic resonance spectroscopy and enzyme assays were performed in Wistar rats 12 weeks after coronary artery ligation. Infarcted rats were allocated randomly to either 6 weeks of training or non-training. Spectra were collected from the calf muscles during sciatic nerve stimulation at 2 Hz. Fibre typing and enzymatic assays were performed on the muscles of the contralateral non stimulated leg. Post-mortem rats were also divided into severe and moderate heart failure according to the lung weight per body weight. At 200 g twitch tension, phosphocreatine and pH were found to be significantly lower in the non-trained severe heart failure group compared with the other groups. Phosphocreatine recovery half-time was significantly longer in the non-trained group with severe heart failure and correlated with the citrate synthase activity in the muscle. The training did not induce a change in the enzyme activities in the infarcted animals with moderate heart failure but did correct the lower citrate synthase activity in the non-trained severe heart failure animals. This normalization of muscle metabolism was achieved by training without any change in calf muscle mass, making atrophy unlikely to be the sole cause of the metabolic changes in heart failure. Training in rats with severe heart failure can reverse the abnormalities of skeletal muscle metabolism, implicating decreased physical activity in the aetiology of these changes. PMID- 7484171 TI - Differences in binding of platelet factor 4 to vascular endothelium in vivo and endothelial cells in vitro. AB - The binding of fluorescein-labelled recombinant human platelet factor 4 (rhPF4) to the vasculature of the hamster cheek pouch in vivo was compared with that to cultured endothelial cells (EC) from human umbilical veins (HUVEC) and arteries (HUAEC) and from human aorta (HAEC). In vivo data: systemically injected rhPF4 rapidly disappeared from plasma in a biphasic pattern (t1/2 = 2 and 41 min). High intensity non-uniform binding of rhPF4 occurred at short specific sites along both arterioles and venules. The length of the intense sites was 76 +/- 46 microns and their frequency was 10 +/- 4 per cm2 cheek pouch. Heparin was injected at 4 and 9 min, but not 30 min, post-rhPF4 displaced most of the high intensity labelling indicating internalization with time. Neither pretreatment with more than 50-fold excess of unlabelled rhPF4 nor histamine- or LTB4-induced vascular macromolecular leakage changed the frequency of short intense sites. In vitro data: uniform time-dependent intense binding of rhPF4 occurred in a similar fashion in subconfluent HUVEC, HUAEC and HAEC. All cell types showed nuclear staining, demonstrating internalization. When heparin was given to EC prior to rhPF4, binding was delayed in time but not blocked. In conclusion, rhPF4 does not bind uniformly with high intensity along pre- and post-capillary vessels of the hamster cheek pouch in vivo as predicted by the rhPF4-labelling of subconfluent (migrating/proliferating) human EC in vitro. The short infrequent sites of intense rhPF4-labeling in vivo may represent regions of endothelial cell migration/proliferation similar to subconfluent EC in culture. PMID- 7484172 TI - Effect of pre-treatment with desferrioxamine and mannitol on radical production and kidney function after ischaemia-reperfusion. A study on rabbit kidneys. AB - The effects of pre-treatment with mannitol and the iron chelator desferrioxamine on oxygen radical formation and glomerular and tubular function after ischaemia in the rabbit kidney were studied. Radicals were measured with ESR and spin trapping. At reperfusion after 60 min of renal ischaemia there was a significant increase in the production of free radicals in the venous effluent from the kidney. Administration of either mannitol or desferrioxamine given before ischaemia and before recirculation reduced the radical production significantly. The iron chelator appeared to be more effective. Glomerular function measured 48 h after reperfusion was significantly better after pretreatment with desferrioxamine and mannitol compared with mannitol alone. Tubular function did not differ between the two pre-treatment groups. PMID- 7484173 TI - Electrically evoked torque-velocity characteristics and isomyosin composition of the triceps surae in young and elderly men. AB - The electrically evoked isokinetic torque-velocity relationship of the triceps surae of eight elderly and four non-trained young men was examined in relation to the isomyosin composition of the soleus and the gastrocnemius muscles, determined under non-denaturing conditions using pyrophosphate gel electrophoresis. The angle specific torque-velocity properties of the triceps surae were measured using maximal percutaneous electrical stimulation at 50 Hz and a release technique. The elderly subjects generated significantly (P < 0.05) less absolute torque at all angular velocities when compared with the young subjects. When the isokinetic data were normalized to the isometric torque, the lower normalized torques generated by the elderly subjects were not statistically different from the young. The total fast isomyosin (FM) content of the soleus and gastrocnemius in the elderly subjects was 22 +/- 13 and 35+/- 18%, respectively. This compared with 29 +/- 8 (n.s) and 44 +/- 8% (n.s.) in the young subjects. When the gastrocnemius and soleus muscles were given an equal weighting and considered together to represent the whole triceps surae, the normalized torque at the fixed angular velocity of 5 rads s-1 was significantly associated with %FM (r = 0.90, P < 0.01), and the isomyosin bands %FM1 (r = 0.90, P < 0.01) and %FM2 (r = 0.93, P < 0.001) when only the elderly subjects were considered. No relationships were observed between contractile characteristics and contractile protein profile when only the young subjects were considered. This was despite the inclusion of a further two sprint and three endurance trained athletes to increase the range of contractile characteristics and differences in muscle composition. PMID- 7484174 TI - Effects of endurance training on left ventricular performance: a study in anaesthetized rabbits. AB - Endurance training is known to increase ventricular performance during exercise and to decrease resting heart rate. The aim of this study was to evaluate a model for endurance training in rabbits and to study the effects of endurance training on local myocardial performance in the left ventricle during resting conditions. One group of rabbits underwent a 10-week exercise training programme. The rabbits trained 5 days a week on a treadmill. Training periods increased gradually from 15 min to 1 h with increments in speed from 0.5 to 1.2 km h-1. After the training programme the rabbits were anaesthetized and studied as acute open-chest preparations. A micro-tip pressure transducer was introduced via apex to the left ventricle and two pairs of ultrasonic crystals were implanted in the left anterior wall to measure segment lengths. One pair measured shortening in the circumferential direction whereas the other pair measured shortening in the longitudinal direction. Heart rate was lower in the trained group (n = 5), 172 +/ 9 beats min-1 (mean +/- SEM), compared with 235 +/- 19 beats min-1 in the control group (n = 8) (P < 0.02). Stroke volume, measured by radio nuclidelabelled microspheres, was greater in the trained rabbits compared with controls (P < 0.03). Shortening in both segments was of similar magnitude for the trained and control groups. End-systolic pressure-length relations (ESPLR) obtained by occlusion of the descending aorta (balloon catheter) showed reduced slopes for longitudinal segments in the trained group compared with the control group (P < 0.05). We conclude that this endurance training programme in rabbits can be used to study myocardial effects of endurance training. Furthermore, the less steep slope of ESPLRs for the longitudinal segment in the trained animals might indicate a structural myocardial remodelling and increased contractile reserve that might be recruited during adrenergic stimulation in the trained group. PMID- 7484175 TI - Endothelin infusion reduces hypoxic pulmonary hypertension in pigs in vivo. AB - Previous work has shown that the plasma levels of the potent vasoactive peptide endothelin (ET) are increased in pathophysiological conditions with increased pulmonary vascular resistance and it has been speculated that ET may play some part in hypoxic pulmonary hypertension. We have therefore evaluated the effects of ET-infusion in the porcine pulmonary circulation after hypoxia-induced hypertension. Pits under general anaesthesia were artificially ventilated through an endotracheal tube and hypoxia was induced by decreasing the fraction inhaled O2 from 0.21 to 0.10. Haemodynamic parameters were continuously recorded using a Swan-Ganz catheter in combination with thermodilution for cardiac output measurements. ET-1 or ET-3 was given as an i.v. infusion through the Swan-Ganz catheter in the right ventricle. Hypoxia induced a reproducible increase in pulmonary vascular resistance (PVR), mean pulmonary artery pressure (MPAP) and right ventricular stroke work (RVSW) while the systemic vascular resistance (SVR) slightly decreased. Cumulative infusion of ET-1 (10, 25 and 50 ng kg-1 min-1) dose-dependently decreased MPAP and PVR; at a higher dose (100 ng kg-1 min-1), the PVR returned to the level observed at hypoxia. ET-infusions at 50 and 100 ng kg-1 min-1 evoked an increase in SVR and a decrease in cardiac output (CO) and stroke volume (SV). RVSW also gradually decreased during ET-1 infusion. Infusion of ET-3 evoked effects similar to those of ET-1 infusions, although the response to ET-3 was not that rapid in onset. In a second series of animals, repeated 15 min periods of hypoxia evoked a stable, reproducible response with a consistent increase in PVR, MPAP and RVSW which returned to baseline values during normoxia. Infusion of ET-1 (25 ng kg-1 min-1) evoked a rapidly developing decrease in PVR and MPAP which was quickly normalized upon cessation of the ET-infusion. ET-1 infusion at this concentration did not per se influence the haemodynamic parameters during normoxia. It is concluded that in the pig, short-term ET infusion reduces the pulmonary hypertension associated with acute hypoxia. PMID- 7484176 TI - Cardiovascular response to exercise in humans following acclimatization to extreme altitude. AB - The purpose of this study was to assess the effects of acclimatization to extreme altitude on the cardiovascular system, using vagal and adrenergic blockade and acute restoration of normoxia during exercise to maximum with one and two legs. Fourteen climbers on an expedition to the Himalayas were studied at a lower base camp (5250 m) following 56-81 days at altitudes between 5250 and 8700 m. After acclimatization, peak heart rate (HRpeak), oxygen uptake (VO2peak) and noradrenaline (NA) were similar during maximal one- and two-legged cycling, whereas peak plasma lactate was higher during the one-legged protocol. HRpeak (range 113-168 beats min-1) was lowest when subjects returned from the higher camps. The degree of partial restoration of HRpeak to more normal values within seconds of 60% O2 inhalation (range 5-35 beats min-1 HRpeak increase) was greatest in subjects with low HRpeak. HR responses to beta-1 blockade increased as a function of HRpeak and the HR responses to atropine were the least in subjects with high HRpeak. These findings suggest that (a) the reduction in HRpeak is linked to the duration and severity of the hypoxaemia, (b) the degree of restoration of HRpeak with acute normoxia is dependent on the level of attenuation or down-regulation of cardiac sympathetic activation (SNA), (c) cardiac vagal drive is masked to a lesser extent in chronic hypoxia because of attenuated SNA and lower HRpeak values, and (d) the lower blood lactate levels at altitude is a function of muscle mass involvement rather than adrenergic activation, as normal peak values were reached during exercise with a small muscle mass. PMID- 7484177 TI - Inhibition of leucotriene-synthesis improves dorsal skin flap survival in the rat. PMID- 7484178 TI - [ACTA: a name, three passions, forty years]. PMID- 7484179 TI - [One more voice]. PMID- 7484180 TI - [The places of mind]. AB - The Sciences of the Mind have gained specific fields in the knowledge upgrading achieved by the relevant scientists and theorists. Three are the best known types of classification, the ones with religious origins excluded: the one of the Mediterranean Ancient civilization, focused on the faculties of the soul; the naturalistic approach based on the complexity levels of the matter; and the bipolar one which separates the Social Sciences from the Natural ones. The places that Man conceives for the Psychological Sciences reflect its ideological status upon the Nature of the Mind. PMID- 7484181 TI - [Thanatotherapeutics]. AB - Thanatotherapeutics, as "ars bene moriendi", belongs to those services that modern medicine should restore and value most. Its exercise demands examining the plurality and heterogeneity of the discourses about death: the biological, the anthropological, the sociological and the theological, among others. Each one of them has its own context of validity and a proper scope. The association between aging and dying permits a discussion of the disablement process and of social death, as paradigms for the most general features of dying. As a concrete and individual process, death occurs nowadays in hospitals and hospices, this fact being essential for examining it in terms of the actors and agents present in this institutional setting. Some principles are derived and discussed, among them three dimensions deemed relevant for appraising thanatotherapeutic work: what is proper according to art, what is good for the individuals concerned and what is just for the society to which they belong. PMID- 7484182 TI - [New concepts relating to histochemistry of the serotonergic neural systems of the raphe nucleus]. AB - The serotoninergic neuronal systems of the brain stem are involved in several processes, like sleep, anxiety and depression. Because of this, these systems have received a great deal of attention during the last few years. As a result, the raphe nuclei have been shown to contain a variety of substances in addition to serotonin. For example they were shown to contain GABA, noradrenaline, enkephalin, somatostatin, substance P and cholecystokinin. Additionally, neuropeptide Y and tirotrophine releasing factor have been found to colocalize with serotonin in the dorsal raphe nuclei. All these results have expanded our knowledge on the raphe nuclei and suggest that many other substances, apart from serotonin, could be involved in the regulation of processes such as sleep, anxiety and depression. Further experiments are necessary to test if this hypothesis is correct. PMID- 7484184 TI - [The psychological meaning of the female friend among Mexican adolescents: a gender comparison]. AB - This study constitutes a continuation of a series of approximation to the knowledge of an adolescent stage. The purpose of this paper is to explore the way in that the females and males adolescents conceptualize a "female friend". With this in mind, we analyzed the information with the semantic network technique in 160 adolescents: 80 females and 80 males divided in two groups of analysis: 8th and 12th school years in private schools from Mexico City. The results permit to observe some differences in their conceptualizations by gender and educative level; but also we observed similarities in the way of to conceptualize a "female friend". Perhaps these findings derived from a cultural tradition that influence the social interactions and by an evolutive stage: the adolescence, when the establishment of the psychosocial identity is an important need and demand. PMID- 7484183 TI - [Variations of the frequency of the diagnosis of depression in Chile]. AB - Different publications have found an increase of depression diagnostic during last years. In this work chilean information available for the last decade in different hospitals (psychiatric, no psychiatric, public-private) is reviewed. Results confirm an increase of relative frequency of depressive disorders according to different classifications systems. Sex differences and possible explanations for this variation in diagnostic frequency are analyzed. Clinical practice would be the principal variable in this modifications. PMID- 7484185 TI - [Combined therapy with tricyclic and MAOI antidepressants in the treatment of resistant major depression]. AB - This paper shows the results of associating tricyclic and MAOI antidepressants in the treatment of resistant major depression. Forty five patients from private practice with diagnosis of major depression according to DSM III R criteria, with negative response to separate tricyclic or MAOI treatment, were given both types associated. They improved without dangerous side effects. PMID- 7484186 TI - [Consideration to eliminate K as a factor of correction in the Mini-Mult 82]. AB - In this study, a discussion and procedure to eliminate K as a factor of correction in the MINI-MULT 82, is presented. This test was developed in Costa Rica and based on Kincannon's Mini-Mult, a short form of the Minnesota Multiphasic Personality Inventory (MMPI). However, the test has been independent from the MMPI, and it is not used as s predictor form. In consequence, the influence of factor K has been considered, either as a suppressor variable or as a contributor to improve the validity of the test. Two samples, one of non patients and other of psychiatric patients were compared using Student's t-test. To do that, the test was scored first, without the K factor and finally, using that one. The results showed statistical differences between the samples (p < 0.001) and they were stronger when the K factor was not used. This findings support the elimination of K as a factor of correction in the MINI-MULT 82. Implications and guidelines about future research, are discussed at the end. PMID- 7484187 TI - [Religious affiliation and mental health in Brazil]. AB - Religiosity is a complex and fundamental socio-cultural phenomenon. Its possible positive or negative influence on the ethiology and treatment of mental illness remains controversial. Evangelical sects, specially the Pentecostals, have expanded dramatically in the last 40 years, in Latin America. Until now, the socio-cultural implications of this process have not been systematically studied. In the present study a group of patients admitted to a psychiatric unit in a general hospital in Campinas, Brazil, was investigated. Diagnosis distribution and length of hospital stay was related to religion affiliation. More functional psychosis and a shorter length of stay was found in the Pentecostal group. Possible implications of these findings are critically discussed. PMID- 7484188 TI - [Organic mental disorders and AIDS]. AB - The mental health professional should be aware of any organic mental disorder in the HIV-seropositive patient, since they are more exposed to this mind of disorders than the general population. The purpose of this study is to provide general guidelines for the early detection of these disorders, giving information on their causes and deeping in the date available on the AIDS-related complex (organic mental disorder specifically related with AIDS). PMID- 7484190 TI - [Sciences of life and sciences of paper]. PMID- 7484189 TI - [Psychiatry in colonial Mexico]. AB - This article describes the knowledges, descriptions and treatments of the mental illnesses by the physicians of the Colonial Mexico (XVI to XIX Centuries). In the first part we can appreciate the spanish ascendancy in the XVI century; in the second one the influence in our country of the Malleus Maleficarum and in the third one the renovation of the Mexican medicine who tries to be part of the universal movement, although in this case, the universe is practically France in the XVIII century and the beginning of the XIX. PMID- 7484191 TI - Abulia and impulsiveness revisited: a conceptual history. AB - Historical analysis suggests that the decline and fall of the will was due not to any major piece of empirical work demonstrating that the concept was unsound but to general changes in philosophical fashion, and to the temporary influence of the anti-mentalistic tenets of behaviorism and the anti-volitional assumptions of psychoanalysis. Clinical disorders like abulia and impulsiveness share conceptual features that 19th-century alienists captured well in their clinical category of disorder of the will. Current accounts, which include semi-explanatory concepts such as "drive", "motivation" or frontal lobe "executive" are not conceptually better than the old notion of will nor are they superior as correlational variables for neurobiological studies. It is suggested that the will, updated according to modern work in the philosophy of action, be re-adopted as a research category in current psychiatry. PMID- 7484192 TI - Factorial structure of the 17-item Hamilton Depression Rating Scale. AB - The Hamilton Depression Rating Scale was applied to 60 depressed inpatients diagnosed using the Composite International Diagnostic Interview. The information to rate the scale was obtained with a semistructured interview to standardize the scale administration method. Items were factorized using principal components analysis with Varimax rotation. Three factors were obtained with the simulation method, accounting for 47% of variance. The first includes the core symptoms of depression. The symptoms of patients having an isolated mood disorder were compared with those having comorbidity with other diagnoses. The comorbidity did not affect the first factor but modified the second factor (anxiety) and the third factor (insomnia). PMID- 7484193 TI - Gender, premorbid characteristics and negative symptoms in schizophrenia. AB - The interrelationships among gender, premorbid functioning, and negative symptoms were examined in a first-admission inpatient sample with DSM-III-R schizophrenia. Fifty-two subjects were assessed with the Schedule for the Assessment of Negative Symptoms (SANS) at baseline and 6-month follow-up. Three indicators of premorbid functioning were examined: the Premorbid Adjustment Scale, the Quick Test, and the GAF for the best month in the year prior to the baseline interview. Men and women had relatively similar ratings on each of the 5 SANS global subscales at both times; they were also relatively similar on most of the indicators of premorbid functioning. The men and women were categorized into low vs moderate high negative symptom groups at baseline, and no differences in premorbid functioning were detected. When the sample was classified into those with and without consistent negative symptoms at baseline and 6-month follow-up, the enduring negative men and women had significantly poorer premorbid functioning in several areas than the consistently non-negative patients. Our findings support the importance of assessing negative symptoms longitudinally and suggest that gender is not strongly associated with negative symptoms and premorbid functioning in patients ascertained at early stages of schizophrenia. PMID- 7484194 TI - Analysis of positive and negative symptoms in schizophrenia. A study from a population of long-term outpatients. AB - Sixty-seven schizophrenic chronic outpatients (DSM-III-R criteria) were examined to study the relationship between symptoms through intercorrelational and factor analysis. Three factors were identified and associated with clusters of symptoms: factor 1 (affective flattening, alogia, avolition-apathy and anhedonia asociality), factor 2 (formal thought disorder and bizarre behaviour) and factor 3 (hallucinations and delusions). Strong associations between factor 1 and high functional impairment were found. In this population, the segregation of the symptoms could fit into a three-syndrome model, these results agree with findings obtained from samples in which acute symptoms predominated. PMID- 7484195 TI - Training in evaluation of expressed emotion using the Japanese version of the Camberwell Family Interview. AB - To evaluate the effect of training in rating expressed emotion (EE) using the Japanese version of the Camberwell Family Interview (CFI), interrater reliability between a certified rater and a trainee was examined. The material was 65 CFI interviews with the families of 46 schizophrenic patients. The two raters independently rated EE status, critical comments (CC), hostility (H), emotional overinvolvement (EOI), warmth (W) and positive remarks (PR). The interrater reliability was satisfactory with regard to EE status, CC, H, EOI and PR. In the category of W, however, the results were unsatisfactory, as there was no positive development of agreement over time. In fathers, the interrater reliability of W was lower than in other relationships. Use of interview transcripts could contribute to a more satisfactory interrater agreement in EOI, and this is recommended as a useful training tool in EE evaluation. PMID- 7484196 TI - Psychological profile of patients suffering from Crohn's disease and ulcerative colitis. AB - Patients with Crohn's disease (CD, n = 61) and ulcerative colitis (UC, n = 24) were studied using an interview and a series of percept-genetic tests describing anxiety, defensive strategies, and the self-image. Attention was specifically directed at a subgroup of CD (n = 15) with its inflammation limited to the colon (colon Crohn, CC). There was also a control group (n = 43). CC had more immature parent relations than the other groups. They were more childishly concrete and more alexithymic. While CC differ from other Crohn patients in many respects, the real cleft opens up with regard to UC, in spite of location of their illness. CC patients obviously have a psychological profile of their own. PMID- 7484197 TI - Risk factors for post-stroke depression. AB - An unselected cohort of 285 stroke patients, median age 69 years, were studied for correlation between potential risk factors and the 1-year incidence of post stroke depression (PSD). The following factors correlated significantly with PSD: a history of previous stroke, a history of previous depression, female gender, living alone and social distress prestroke. Further, social inactivity, decrease in social activity, pathological crying and intellectual impairment at 1 month but not functional outcome correlated to PSD. A multivariate regression analysis showed that intellectual impairment explained 42% of variance of mood score. Major depression was unrelated to lesion location. We conclude that etiology to PSD is a complex mixture of prestroke personal and social factors, and stroke induced social, emotional and intellectual handicap. PMID- 7484198 TI - Cost-effectiveness of clozapine treatment in therapy-refractory schizophrenia. AB - The costs and effects of clozapine treatment of refractory schizophrenic patients have been discussed recently. This study shows that 18 months of clozapine treatment results in an improvement of symptoms and social functioning in approximately 70% of treatment-refractory schizophrenic patients, compared with treatment with conventional neuroleptics during a similar period of time. Treatment with clozapine reduces the cost of inpatient care but places increased demands on active rehabilitation resources in outpatient care. This leads to increased total costs in a short-term perspective, but clozapine treatment is cost-saving for annual maintenance therapy. These costs must be weighed against the positive effects on psychotic symptoms and social functioning. PMID- 7484199 TI - The Nordic comparative study on sectorized psychiatry. II. Resources of the psychiatric services and treated incidence. AB - As a part of a Nordic comparative study on sectorized psychiatry, treated incidence was related to the resources and dynamic qualities of psychiatric services in 7 catchment areas. One-year treated incidence cohorts were used. Data was collected concerning number of beds and staff, number of long-term patients and turnover rate of patients in the services and availability of specialized services. A positive correlation was found between rates of outpatient staff and treated incidence. No statistically significant correlation was found between the dynamic qualities of the services and treated incidence. Treated incidence of dependence was the highest in a center that had a special service unit for abusers. Special services for young and old people were not clearly reflected in treated incidence in respective patient groups. PMID- 7484201 TI - Timing of completed suicides among residents of Olmsted County, Minnesota, 1951 1985. AB - Previous studies of suicide timing may have been biased by incomplete ascertainment of suicides and by delays between the suicidal act and subsequent death. Those potential biases were assessed and minimized in this population based study by using the unique resources of the Rochester Epidemiology Project in Olmsted County, Minnesota. Using these more accurate data, we confirmed previous reports of no excess suicides on birthdays (+/- 3 days), or during 3 United States national holidays. While most prior reports found excess post holiday suicides and suicide peaks on Mondays, those findings were not observed in Olmsted County. Because 93% of deaths occurred on the date of the suicidal act, using date of death instead of the actual date of suicide is sufficient for most research purposes. PMID- 7484200 TI - The association between economic and social factors and mental health in Finland. AB - The aim of the study was to examine the social and economic variables associated with mental health by means of telephone interviews of a randomly chosen population sample (n = 1557) in Finland in 1993. Mental health was evaluated by means of a 12-item General Health Questionnaire. A mental disorder, indicated by the GHQ score, was detected in 18.3% of the interviewees. Mental disorders were more common in women than in men. Other factors associated with a mental disorder were unemployment, financial difficulties and insufficient social support from other people. Mental disorders, indicated by the GHQ score, were common in the long-term unemployed, users of hypnotics and sedatives, and in those who were uncertain about their future. In stepwise logistic regression analyses the statistically independent associates with impaired mental health were suicidal thoughts, receiving of housing allowance and female sex. On the other hand, being on salary was a protective factor. PMID- 7484202 TI - Lithium addition to neuroleptic treatment in chronic schizophrenia: a randomized, double-blind, placebo-controlled, cross-over study. AB - We studied the effect of lithium addition to neuroleptic treatment in chronic schizophrenia, for which contradictory results have been produced in previous studies. Twenty-one chronic schizophrenic inpatients received lithium in a study with randomized, double-blind, placebo-controlled, cross-over design consisting of 8 weeks each of treatment with lithium capsules and identical placebo capsules. The total Brief Psychiatric Rating Scale (BPRS) scores at week 8 of the lithium treatment were improved significantly compared with those at week 8 of the placebo treatment. Of the BPRS subscales, however, only anxiety-depression improved, whereas none of the subscales for anergia, thought disturbance, activation and hostile-suspiciousness improved. There was no significant difference between the total Scale for the Assessment of Negative Symptoms (SANS) scores at any time during lithium and placebo treatment. These results suggest that the addition of lithium to neuroleptic treatment improves anxiety-depression in chronic schizophrenia. PMID- 7484203 TI - Coercive measures in psychiatric care. Reports and reactions of patients and other people involved. AB - In ongoing studies of the quality of the mental health services in two Swedish counties, two thirds of the committed versus about one third of the voluntarily admitted patients reported coercive measures during the index period of care. Committed patients reported an equal degree of coercive treatment and restraint, whereas restraint dominated among the voluntarily admitted. The majority of the patients described the coercive measures as implemented by fait accompli; force was reported in 23% of the examples given by the committed patients but never by the voluntarily admitted. The committed patients justified 19% and the voluntarily admitted 38% of the coercive measures reported; the committed patients justified coercive treatment and restraint to about the same extent; the voluntarily admitted patients justified 65% of the examples of restraint but only 20% of the examples of coercive treatment. There was a 70% concordance between the reports of the committed patients and psychiatric personnel as to the occurrence of coercion, but the head nurses tended to state that treatment had been implemented by persuasion in cases where the patients stated that implementation was by coercion. PMID- 7484204 TI - Appearance, body image and distress in facial dysmorphophobia. AB - Compared with surgical patients awaiting rhinoplasty and control subjects, dysmorphophobic patients were more dissatisfied with facial appearance, more anxious, more depressed, more neurotic and less extrovert. Morphanalysis, an objective measure of facial appearance, was applied to the first 11 dysmorphophobic and first 11 surgical subjects. The dysmorphophobic group had a variety of abnormal features that were not identified by medical practitioners or the patient. These subtle anomalies were often unrelated to the focus of dissatisfaction which was usually normal. A panel of lay judges rated the appearance of the dysmorphophobic group as being intermediate between the control and surgical groups but not significantly different from either. PMID- 7484205 TI - Successful estradiol treatment of psychotic symptoms in the premenstrual phase: a case report. PMID- 7484206 TI - Effect of paroxetine on seizure length during electroconvulsive therapy. AB - This study reports the effect of paroxetine, a selective serotonin reuptake inhibitor, on seizure length during electroconvulsive therapy. Seven inpatients taking paroxetine were compared with 7 controls taking tricyclic antidepressants. The mean seizure length of the paroxetine group was approximately twice that of the control group. PMID- 7484207 TI - MR imaging of contrast-enhanced porcine myocardial infarction. Assessment of reperfusion and tissue viability. AB - To assess the usefulness of Dy-DTPA-BMA-induced signal reduction, as an indicator of myocardial viability, myocardial infarction was induced in 17 domestic pigs by ligating a diagonal branch of the left anterior descending coronary artery (LAD). In 6 pigs, Dy-DTPA-BMA (1 mmol/kg b.w.) was administered 4 hours after induction of ischaemia. In 5 additional pigs, Gd-DTPA-BMA (0.3 mmol/kg b.w.) and Dy-DTPA BMA (1 mmol/kg b.w.) were simultaneously injected after 4 hours of ischaemia to ascertain whether Dy-DTPA-BMA counteracted the signal enhancement effect of Gd DTPA-BMA. A further 6 pigs with infarctions, not administered contrast medium, served as controls. All pigs were sacrificed after 6 hours of ischaemia, and the extirpated hearts were investigated with MR (ex vivo). The concentrations of Dy and Gd were determined in tissue samples from infarcted and non-ischaemic myocardium. The extracellular concentrations of both contrast media were monitored over time during 2 hours in the double-contrast group (in vivo), using a microdialysis technique and analysed by inductively coupled plasma atomic emission spectrometry (ICP-AES). The infarctions demonstrated a high SI in the proton density- and T2-weighted sequences, in both the Dy-DTPA-BMA and control groups, although the former group demonstrated a 3-fold greater concentration of Dy in infarcted compared with non-ischaemic myocardium. Dy-DTPA-BMA did not counteract the Gd-DTPA-BMA-induced enhancement of the infarcted tissue despite a 3-fold higher concentration. This lack of detectable susceptibility effects of Dy may be caused by a loss of cell membrane integrity in the infarcts, resulting in a homogeneous intra- and extracellular distribution of the contrast agent. This hypothesis of an expanded volume of distribution in infarcted tissue was further supported by the microdialysis data, demonstrating a similar extracellular concentration of contrast agents in infarcted and non-ischaemic myocardium, despite a proven 3-fold greater concentration in infarcted tissue samples. To investigate whether Gd-DTPA-BMA-enhanced MR imaging (ex vivo) permits differentiation between reperfused and non-reperfused myocardial infarction, and whether Dy-DTPA-BMA-enhanced MR imaging enables a differentiation between reversible and irreversible myocardial injury following reperfusion, myocardial infarction was induced in 24 domestic pigs (divided into 4 groups) by placing a patched ligature around a diagonal branch of the LAD. Four additional hearts were reperfused after 2 min of brief occlusion, not long enough to cause irreversible injury.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7484208 TI - Percutaneous automated nucleotomy and imaging with CT, MR and diskography in lumbar disk herniation. PMID- 7484210 TI - Nerve growth factor treatment of adult rats selectively enhances innervation of urinogenital tract rather than vascular smooth muscle. AB - Following treatment of adult rats with nerve growth factor (0.5 mg/rat, three times a week for 3 weeks), the innervation of cardiovascular and urinogenital tract smooth muscle was investigated using immunoassay and immunohistochemical techniques. Substance P and calcitonin gene-related peptide levels were increased in the vas deferens, but not in the atria or femoral artery. Neuropeptide Y and vasoactive intestinal polypeptide levels were unchanged. In penile tissues, there was a marked increase in the density of substance P-, calcitonin gene-related peptide-, neuropeptide Y-, tyrosine hydroxylase- and vasoactive intestinal polypeptide-containing nerves innervating the urethra and in SP-containing nerves in the tunica with little changes in the innervation of the deep dorsal vein and artery and corpus cavernosum. In the bladder, there was increased innervation of the detrusor by neuropeptide Y- and vasoactive intestinal polypeptide-containing nerves, but a decrease in innervation by substance P-containing nerves in the trigone. There were no changes in the density of innervation of the femoral artery after nerve growth factor treatment. Thus, in the mature rat, sensory and sympathetic nerve innervating urinogenital tract smooth muscle appear to be more responsive to exogenous nerve growth factor than those innervating cardiovascular smooth muscle. This may reflect an ongoing requirement of plasticity of innervation in the urinogenital tract of the sexually mature animal. PMID- 7484212 TI - Differential distribution of GABAA receptor subunits in soma and processes of cerebellar granule cells: effects of maturation and a GABA agonist. AB - Quantitative analysis of the density of alpha 1 and beta 2/3 GABAA receptor subunits was performed at the electron microscope level after indirect pre embedding immunogold labeling with subunit-specific antibodies of rat cerebellar granule cell cultures grown for 4 or 8 days and in the presence or absence of the GABAA receptor agonist 4,5,6,7-tetrahydroisoxazolo[5,4c]pyridin-3-ol (THIP). THIP (150 microM) induced a 2-fold increase in the number of alpha 1 and beta 2/3 subunits in both cell bodies and processes in 4-day-old cultures. Extending the culture period to 8 days led to a polarization of the receptor expression, since the increase in the number of subunits selectively was observed in the processes. Moreover, a general subcellular differentiation of the receptor population was observed in all culture conditions, since the ratio between the two subunits (beta 2/3; alpha 1) was four times higher in cell bodies compared to processes. A detailed analysis of the less mature (4-day-old) cultures revealed the existence of two populations of neurons exhibiting differences in the average number of receptors. During maturation neurons with few receptors developed into cells with a higher density of receptors resulting in a single population of the latter neurons, a process enhanced by exposure to THIP. This may indicate that receptor development is a discontinuous process with individual neurons following different temporal patterns. In double-labeling experiments, a spatially close association of the alpha 1 and beta 2/3 subunits could be seen, but the subunits were more frequently found separated from each other. In spite of the fact that exposure of the neurons to THIP increased the total number of receptor subunits, its presence apparently prevented formation of receptors with this subunit composition. Interestingly, receptor subunit clusters, consisting of alpha 1 alone, were more frequently observed than composite (alpha 1; beta 2/3) clusters. This substantiates the view that receptors not having alpha 1 and beta 2/3 subunits in the same complex may exist. PMID- 7484209 TI - Plasticity in the myenteric plexus of the rat ileum after long-term sympathectomy. AB - To investigate the effect of chronic sympathectomy on the innervation of a tissue with an extensive intrinsic component, 1-week-old rat pups were treated with 50 mg/kg guanethidine for 3 weeks, a treatment shown to produce complete and long lasting sympathectomy, and the ileum examined. Changes in the levels of noradrenaline, neuropeptide Y, calcitonin gene-related peptide, substance P and vasoactive intestinal polypeptide in the external muscle layers containing the myenteric plexus of the ileum were determined between 6 and 20 weeks of age. After sympathectomy, noradrenaline levels were initially depleted (3% of age matched controls at 6 weeks, P < 0.001, and 18% of age-matched controls at 12 weeks, P < 0.001), but were not significantly reduced at 20 weeks (67% of age matched controls). Such increases in noradrenaline content with time after sympathectomy did not occur in the mesenteric vein (levels in 20-week-old sympathectomized rats were 2% of the control values (P < 0.001). In the myenteric plexus, catecholamine fluorescent nerve fibers were seen in the 12-week-old sympathectomized rats, although tyrosine hydroxylase-immunoreactivity was absent. Guanethidine sympathectomy had no effect on the neuropeptide levels in 6-week-old rat ileum but there was a selective increase at 20 weeks; the levels of calcitonin gene-related peptide and substance P were increased (X3, P < 0.001 and X1.6, P < 0.05, respectively) while vasoactive intestinal polypeptide and neuropeptide Y levels were unchanged. Short-term sympathectomy (destruction of sympathetic nerve terminals by acute 6-hydroxydopamine treatment) had no affect on noradrenaline or peptide levels in this tissue.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7484211 TI - Existence of a putative specific postsynaptic density protein produced during Purkinje cell spine maturation. AB - This study identified a 140 kDa polypeptide as a putative specific component of Purkinje cell spines' postsynaptic densities and which began to appear during the critical period of cerebellar cortex synaptogenesis. Mouse cerebellar cortices at postnatal days 5, 7, 9, 11, 15 and young adult, between days 30 and 40, were used to purify subcellular fractions of synaptosomes, synaptic membranes and postsynaptic densities. The purity of the subcellular fractions was assessed by electron microscopy and the protein composition of the different fractions was characterized by sodium dodecyl sulfate-polyacrylamide gel electrophoresis. Polypeptides of apparent molecular weights of 25, 26, 27, 30, 33, 37, 43, 45, 52, 64, 74, 85, 94, 110, 125, 130, 165 and 174 kDa were found in the synaptosomal fractions of all the ages studied, even before the critical period of synaptogenesis, at postnatal day 7, when the postsynaptic densities were still nonexistent, indicating that the polypeptides are nonspecific constituents of these structures. On the other hand, a 140 kDa polypeptide was detected in the postsynaptic density fractions at postnatal day 11, immediately after postsynaptic structures began to appear, suggesting the possibility that this protein is a specific component of the cerebellar cortex postsynaptic densities. The 140 kDa polypeptide was electroeluted from the gel and analysed for its amino acid composition by reverse-phase high-pressure liquid chromatography. The analysis showed that this protein has a high content of nonpolar amino acid residues, such as leucine, isoleucine, glycine, phenylalanine and valine. A hypothetical model relative to the participation of the 140 kDa protein in the molecular organization of the postsynaptic density is suggested which may contribute to the understanding of the role played by this structure in synaptic function. PMID- 7484213 TI - Embryonic rat motoneurons express a functional P-type voltage-dependent calcium channel. AB - Only L- and N-type high voltage-activated calcium currents (HVA ICa) have been demonstrated in identified embryonic spinal motoneurons. However, pharmacological experiments suggest that other HVA ICa, including P-type, govern neurotransmitter release at the adult neuromuscular junction. We sought to analyse if embryonic motoneurons express these other ICa, using the whole-cell voltage-clamp method on motoneurons purified by a new metrizamide-panning technique from E15 rat embryos. In addition to L-type dihydropyridine-sensitive and N-type omega-GVIA-sensitive currents, motoneurons express two other HVA ICa. One has properties related to the P-type channel currents described in Purkinje cells: it is inhibited by the peptide omega-agatoxin-IVA with a maximal effect at 100-200 nM. The inhibited current has a characteristic sustained component during depolarizing test pulses. Furthermore, 50-100 nM concentrations of omega-agatoxin-IVA reduce the increase in cytoplasmic calcium concentration observed after depolarization. The other HVA ICa is resistant to saturating concentrations of verapamil, omega-conotoxin GVIA and omega-agatoxin-IVA which block L, N and P-type HVA ICa, respectively. These results suggest that it is now possible to dissect, using a simple method of purification, the properties of the ICa in embryonic mammalian motoneurons and to provide pharmacological evidence for multiple calcium channels which may be involved in regulation of their activity during development. PMID- 7484214 TI - Retinoic acid induces cholinergic differentiation of NTera 2 human embryonal carcinoma cells. AB - Retinoic acid (RA), a natural metabolite of vitamin A, influences the survival and neurotransmitter phenotype of several classes of vertebrate neurons during development. We now report that RA induces a subpopulation of NTera 2/clone D1 (NT2) human embryonal carcinoma cells to differentiate into postmitotic cells with cholinergic properties (NT2-N cells). After growth for 6 days in the presence of RA (10 microM) low levels of the acetylcholine-synthesizing enzyme choline acetyltransferase (ChAT) were detected in NT2 cell cultures. ChAT activity in the NT2 cell cultures continued to increase for at least an additional 22 days to a final activity of 50 pmol ACh synthesized/min/mg protein. Immunohistochemical staining of RA-treated cultures demonstrated that only those cells with a neuronal morphology (NT2-N cells) expressed the human ChAT protein. Since such cells comprised a small proportion (approximately 20%) of the population, the ChAT activity per neuronal cell was estimated to approach 250-300 pmol ACh/min/mg protein. Cultures composed of > 95% NT2-N cells had significantly lower ChAT specific activities and this could be increased by either ciliary neurotrophic factor or leukemia inhibitory factor, but not by nerve growth factor. We conclude that NT2 cells provide a system in which to study the molecular events that underlie neurotransmitter choice during the differentiation of human cholinergic neurons. PMID- 7484215 TI - Hydroxy- and non-hydroxy-galactolipids in developing rat CNS. AB - Rat spinal cord (1-24 weeks postnatal) was analysed by HPLC for various species of galactolipids that accumulate in mammalian myelin during development. Cerebral tissue of the same animals was taken as reference. The levels of the major galactolipids, galactosylceramide (GalCer) and its sulfated analog (SGalCer), increased linearly during the first 2 months after birth. At 3 months, constant levels were reached that were approx. 4-fold (GalCer) and 2.5-fold (SGalCer) higher than in cerebral tissue of corresponding age. The accumulation of galactoglycerolipids slightly preceded that of galactosphingolipids. Levels of galacto-glycerolipids were much lower (4% of galactosphingolipids in 3-and 2.5% in 6-month-old spinal cord on weight basis) and decreased upon CNS maturation. During the first postnatal month, the ratio of non-hydroxy- over hydroxy-species (NFA/HFA) of cerebral GalCer declined from 2.2 to 0.5 whereas the NFA/HFA ratio for cerebral SGalCer increased from 1.0 to 1.8 in the same period. Through development the hydroxy-species contributed 56-60% to GalCer and 28-41% to SGalCer in spinal cord, whereas in cerebrum of 24-week-old rats 73% of GalCer and 48% of SGalCer was alpha-hydroxylated in the ceramide moiety. These data point to different developmental programs with respect to galactolipid metabolism of oligodendrocytes in high- (spinal cord) as compared to low-myelinated (cerebral) areas of rat CNS. PMID- 7484216 TI - ATP synthase subunit C storage in the polymorphonucleocytes of late infantile and juvenile batten patients. AB - In late infantile and juvenile forms of neuronal ceroid lipofuscinosis, commonly known as Batten disease (BD). ATP synthase subunit c accumulates in the lysosomes of neural cells. By using polyclonal antibodies, raised against bovine liver subunit c and an image analysis system for the quantification of antibody-linked alkaline phosphatase reaction, we have demonstrated that polymorphonucleocytes (PMN) from a late infantile and a juvenile BD patient stored several-fold more subunit c as compared to normal PMN. PMID- 7484217 TI - Morphological and GABA-immunoreactive development of the embryonic chick telencephalon. AB - The development of neurons utilizing gamma-aminobutyric acid (GABAergic neurons) in prosencephalon and telencephalon from chicken embryonic days 4-14 (E4-E14) was studied by means of immunohistochemistry. Furthermore, routine histology and transmission electron microscopy. respectively, were performed in order to study the morphological development in the designated area. The main finding is that development of GABAergic neurons in the chick telencephalon is rapid; the GABA neurons are appearing in bulk at day 8, being "overexpressed" at days 10-11, decreasing in numbers thereafter and achieving mature morphology on day 14, which is considerably faster than in the rodent. Morphological analysis revealed that the prosencephalon mainly consisted of a thin layer of undifferentiated neuroblasts in the E4 embryo. By E6, the prosencephalon had increased in thickness and occasional cells outside the neuroepithelium showed a more mature morphology with a few cells weakly staining positive for GABA. At E8, the prospective granular and subventricular layers had developed. At E14, the appearance of the telencephalon is approximating that of the adult since both ependymal cells and morphologically mature neurons can be seen. PMID- 7484219 TI - Epidermal growth factor influences the neurotrophic/differentiating action of nerve growth factor. AB - Exposure of naive PC12 cells, sympathetic neurons from rat superior cervical ganglia, and brain-derived septal neurons to epidermal and nerve growth factors simultaneously resulted in some alteration of cellular events induced by nerve growth factor alone. A more pronounced decline of catecholamine content, no additional change in acetylcholinesterase activity, and additive stimulation of RNA and protein syntheses were found in PC12 cells. Earlier elevation of the enzyme activity was observed in sympathetic but not in septal neurons. Epidermal growth factor appeared to support independently the same level of acetylcholinesterase activity in septal neurons as revealed for nerve growth factor during the first week and cell survival throughout 2 weeks of observation. The data obtained indicate that epidermal growth factor augments temporarily some effects of nerve growth factor, thus supporting the idea of an important role of mitogenic growth factors in neural development as complementary and/or substitutive regulators of nerve cell differentiation and survival. PMID- 7484218 TI - Changes in fast axonal transport in sensory neurons during tadpole metamorphosis. AB - Fast axonal transport of radiolabeled protein was examined in lumbar and tail dorsal root ganglion (DRG) neurons at progressive stages of bullfrog tadpole metamorphosis. Accumulation of [35S]methionine-labeled protein proximal to a lumbar peripheral nerve ligature (at a fixed distance from the DRG) increased as tadpoles advanced from premetamorphosis through prometamorphosis to metamorphic climax. The rate of increase was steeper when expressed as a percentage of protein synthesized in the neurons of origin than when expressed as a percentage of total DRG protein synthesis. Further, the increase was not secondary to a rise in protein synthesis. In contrast, fast axonal transport decreased in DRG neurons of the tail at the onset of metamorphic climax, when tail resorption is initiated. The stage-related increase in protein transport in lumbar nerves is due, at least in part, to an increased rate of transport. As determined from optically detected anterograde organelles in individual lumbar nerve axons, an approximate doubling of the fast transport rate occurred between the premetamorphic stage and metamorphic climax. In addition, the rates of organelle transport in lumbar axons of adult bullfrogs were significantly greater than in corresponding axons of tadpoles at metamorphic climax, further suggesting that organelle velocity is a developmentally regulated parameter of fast axonal transport. PMID- 7484221 TI - Community-acquired lower respiratory tract infection in the elderly: a community based study of incidence and outcome. AB - BACKGROUND: We studied the incidence, characteristics, and survival of elderly patients from a defined community who had pneumonia or bronchitis (lower respiratory tract infection). METHODS: This study was a population-based retrospective cohort study of residents of Rochester, Minnesota, aged 65 years or older with a first episode of pneumonia or bronchitis during the calendar year 1987. RESULTS: Overall age- and sex-adjusted incidence rates for an initial episode in a calendar year were 5452 per 100,000: 2420 per 100,000 (95 percent confidence interval, 2056 to 2783 per 100,000) for bronchitis and 3032 per 100,000 (95 percent confidence interval, 2639 to 3425 per 100,000) for pneumonia. After exclusion of eight cases diagnosed at autopsy, the overall 30-day mortality was 10.7 percent. Patients with pneumonia had lower survival than expected for the Minnesota white population (log-rank statistic = 117.38, P < 0.0001). The observed survival of patients with bronchitis was also significantly less than expected (log-rank statistic = 6.25, P = 0.012). CONCLUSION: Lower respiratory tract infections are common in the elderly. Most patients in this study were not hospitalized, and atypical presentations were not observed. Early mortality (30 days) was high. Among elderly patients with either pneumonia or bronchitis, the survival was lower than expected. This study confirmed the need for population based studies, because more than two-thirds of patients would have been missed if only hospitalized patients were included. PMID- 7484222 TI - Vaginal birth after cesarean: a 5-year experience in a family practice residency program. AB - BACKGROUND: The national health objective for the year 2000 is to have an overall Cesarean section rate of 15 percent, a primary Cesarean section rate of 12 percent, and a vaginal birth after Cesarean (VBAC) rate of 35 percent. The current national statistics for the most recent year available, 1991, are 23.5 percent, 17 percent, and 24.2 percent, respectively. This study evaluates a VBAC program at a community-hospital-based family practice residency program. METHODS: This study was a retrospective chart review of 996 family practice service deliveries from 1988 to 1992. RESULTS: Of the 98 patients who had a previous Cesarean section, 87 patients (89 percent) were eligible for a trial of labor. Fifty-six patients (64 percent) accepted a trial of labor and 31 patients (36 percent) refused. The overall Cesarean section rate was 15 percent, the primary Cesarean section rate was 11 percent, the VBAC rate was 44 percent, and the successful VBAC rate was 77 percent. CONCLUSION: We report a successful VBAC program at a community-hospital-based family practice residency program. PMID- 7484220 TI - Taurine deficiency in dissociated mouse cerebellar cultures affects neuronal migration. AB - The role of taurine in the process of neuronal migration was studied in a microwell cell culture system. Immunocytochemical analysis of the cellular composition of this culture system revealed the presence of the astrocytic marker GFAP in some structures such as the aggregates of neuronal bodies and in those cables used for migration, resembling what is described in vivo. The neuronal marker gamma-enolase stained practically all structures, including the aggregates and all cables. The intracellular taurine concentration was reduced by 60% in mouse cerebellar granule cells treated with a blocker of taurine transport, guanidinoethane sulfonate (GES). Under these conditions cell migration was markedly reduced to approximately 50% of that in untreated cultures. Both, taurine depletion and impairment of cell migration induced by GES were prevented by adding taurine to the culture medium. Taurine deficiency similarly affected different morphological parameters such as the number of cables suitable for neuronal migration as well as the number of migrating neurons. The number of aggregates of neuronal bodies was significantly increased, by about 30%, as a consequence of the reduced migration. Taurine alone did not exert any effect on the parameters evaluated. GES treatment of granule cells did not affect mitochondrial metabolism or K(+)-stimulated Ca(2+)-dependent [3H]-D-aspartate release. This suggests that the described effects of taurine deficiency were not due to an alteration of neuronal viability and that the action of GES was not simply due to unspecific and deleterious effects. These results are in agreement with those obtained in in vivo studies. This approach represents a useful model to investigate the role played by taurine in the process of neuronal migration. PMID- 7484223 TI - Physicians' attitudes toward complementary or alternative medicine: a regional survey. AB - BACKGROUND: A survey exploring attitudes toward complementary or alternative medicine was distributed to 295 family physicians in the Chesapeake region. Physicians were asked about their knowledge, usage of, and training in complementary therapies and what evidence they expected of complementary medicine to aid in accepting these therapies. METHODS: Questionnaires were distributed at three separate conferences of family physicians with 180 physicians responding. RESULTS: More than 70 to 90 percent of the physicians considered complementary medical therapies, such as diet and exercise, behavioral medicine, counseling and psychotherapy, and hypnotherapy, to be legitimate medical practices. A majority had referred patients to nonphysicians for these therapies or used some of them in their own practices. Homeopathy, Native American medicine, and traditional Oriental medicine were not favored as legitimate medical practice. Areas where physicians had the least amount of training were most likely to be considered as alternative medicine by them. Seventy percent of responding physicians expressed interest in training in multiple areas of alternative medicine. Additionally, there was a strong positive correlation between evidentiary rules physicians believed should apply to orthodox medicine and to alternative or complementary medicine. CONCLUSIONS: Results of this survey show a high interest in alternative and complementary medicine by physicians. Some therapies are already being used by these physicians, and training is desired in most areas. PMID- 7484224 TI - Dimensions of happiness: a qualitative study of family practice residents. AB - BACKGROUND: Happiness is related to both positive and negative forces. Positive factors, or happiers, that compensate for stressors in residency have only recently been recognized in a pilot study of 14 family practice residents. These happiers are positivism, the pursuit of goals, relationships, a religious belief system, and feedback. The purpose of this qualitative study was to describe happiers of residency in family practice in each postgraduate year and relate the findings to established theories on well-being. METHODS: A qualitative interview with participant observation was used to study 59 residents at four family practice residency sites. Interviews were semistructured and audiotaped. Tapes and field notes were analyzed for trends. The Faces Scale was administered as a happiness visual analog scale. RESULTS: The importance of a well-balanced lifestyle, as well as the pursuit of goals, relationships, and accentuation of the positive, were found to be happiers for all residents interviewed. Religious commitment, feedback, and a sense of control or autonomy were important to the level of happiness of more than 60 percent of residents interviewed. Feedback was more important to 1st-year residents, and a sense of control was more important to 2nd-year and 3rd-year residents. CONCLUSION: The seven happiers described in this study were similar to happiness factors reported in the social science literature. Our findings could contribute to theories of resident well-being and further the research on well-being in residency training. PMID- 7484225 TI - Myxedema coma in the elderly. AB - BACKGROUND: Myxedema coma in the elderly, although uncommon, is frequently overlooked and has a high mortality rate. Signs and symptoms are many and are often insidious. Nearly every organ system is involved. Prompt recognition and treatment are mandatory for a successful outcome. METHODS: A case study is presented. Using the key words "myxedema" with the word "aged," MEDLINE files were searched from 1989 to present. Articles dating before 1989 were accessed from the reference lists of the more recent articles. RESULTS AND CONCLUSIONS: This review describes the signs and symptoms of myxedema coma in the elderly. Epidemiology and histopathology of the disorder are discussed. Prompt recognition and emergency medical treatment are essential for a successful outcome. Prevention requires screening of elderly patients at risk for hypothyroidism and assuring thyroid hormone replacement therapy. PMID- 7484226 TI - Lemierre syndrome: postanginal sepsis. AB - BACKGROUND: Lemierre syndrome, or postanginal sepsis, was first described in the early part of this century and is characterized by pharyngitis, followed by high fever and rigors, cervical adenopathy, thrombophlebitis of the internal jugular vein, distant abscess formation, and icterus, associated with isolation of Fusobacterium necrophorum from blood. METHODS: This report describes a case of postanginal sepsis and reviews the medical literature on postanginal sepsis obtained through the MEDLINE data base using Fusobacterium as the key search word. RESULTS: The features of Lemierre syndrome have changed little since the original description, through the prognosis has improved dramatically since the development of antibiotics. Appropriate management includes prompt administration of an antibiotic with good anaerobic coverage, drainage of persistent abscesses, and continued antibiotic therapy until radiographic resolution of abscess is achieved. CONCLUSIONS: Although Lemierre syndrome is a relatively uncommon disease, the primary care physician needs to be aware of the clinical features and management to treat appropriately. PMID- 7484227 TI - Relation of infant mortality to the availability of maternity care in rural Florida. AB - BACKGROUND: This cross-sectional study was designed to explore the impact of the availability of maternity care services on the infant mortality rates in nonmetropolitan (rural) counties in Florida. METHODS: We evaluated the sufficiency of physicians providing maternity care in each rural county. We then constructed a mathematical model to compare physician availability with the infant mortality rates for each county, while controlling for socioeconomic variables. RESULTS: Thirty-one family physicians and 974 obstetrician gynecologists were delivering babies in Florida in 1991. Forty-seven counties were lacking in maternity care services; 45 of these counties had family physicians who practiced in the county but did not provide maternity care services. There was a negative correlation in rural counties between availability of maternity care services and infant mortality (R = -0.42, R2 = 0.176, P = 0.012), implying that 17.6 percent of the variation in rural Florida's infant mortality was explained by a ranking in physician availability. Multivariate analysis revealed that increasing infant death rates can be predicted by decreasing physician availability (P = 0.003). A multiplicative risk model developed for this study demonstrated that the loss of 1 family physician delivering babies would predict the increase of infant mortality by 2.3 percent, and the loss of 1 obstetrician-gynecologist increased infant mortality by 9.6 percent. CONCLUSIONS: Access to maternity care for women in rural Florida is a problem that could be hampering Florida's ability to reduce its infant mortality rate. Family physicians appear to be the most geographically distributed health care providers in Florida; therefore, strategies should be developed to recruit Florida's rural family physicians into maternity care. PMID- 7484230 TI - Uterine-vesicular rupture during trial of labor. PMID- 7484229 TI - Atrial fibrillation during flexible sigmoidoscopy in a 36-year-old man. PMID- 7484228 TI - Serotonin syndrome in Parkinson disease. PMID- 7484231 TI - Unusual cause of unresponsiveness during the second stage of labor. AB - This case report illustrates an unusual occurrence during the second stage of labor. After appropriate evaluation for life-threatening causes, a psychiatric process must be considered. Although our patient refused further psychological evaluation, we believe that she exhibited a trancelike state during the second stage of labor and suffered from amnesia in which one awakens to the current situation. It is known that certain individuals are predisposed to dissociate in response to stress. According to our patient's history, she seems to fall into this category. Although those involved in obstetric care must remain alert to the possibility of catastrophic events intrapartum, we must also remember to consider unusual psychiatric processes. PMID- 7484232 TI - "But doctor, I'd prefer to have a cesarean section": when public policy conflicts with patient preference. PMID- 7484233 TI - Relationship-centered care: beyond the finishing school. PMID- 7484234 TI - Happy residents, happy people, or both? PMID- 7484235 TI - Management of obesity. PMID- 7484236 TI - Management of obesity. PMID- 7484237 TI - Rapid antigen detection testing. PMID- 7484238 TI - Polypharmacy in nursing homes. PMID- 7484239 TI - Polypharmacy in nursing homes. PMID- 7484240 TI - [The effect of combined toxicity on the development of alcoholic pancreatic lesions. A long-term experimental trial]. AB - The aim of this study was to evaluate the adverse effects on the exocrine pancreas of ethanol and ethanol with congeners which coexist in alcoholic beverages most commonly consumed by the Portuguese population. Eighteen male Wistar rats were divided into three groups and submitted to a daily intraperitoneal injection of a hydroalcoholic solution of ethanol (SHAE) and a hydroalcoholic solution of ethanol, acetaldehyde, methanol and higher alcohols (SHAF); the third group served as a control and received an equivalent volume of an isocaloric solution of dextrose. All the animals were sacrificed at the end of the 9th week of the experiment. The following histological lesions were considered: acinar cell necrosis and steatosis, ductal dilatation, intraluminal plugs, parenchymal inflammation, fibrosis, peripancreatic fat necrosis and inflammation. Their severity was graded by means of a scoring system. The histopathologic changes which characterize pancreatitis (acinar cell necrosis, parenchymal inflammation, fibrosis and peripancreatic fat necrosis and inflammation) were found in the majority of animals of both study groups, but in none of the control group. The lesions tend to be more frequent and severe in the group treated with ethanol and its congeners (SHAF) than in the SHAE group; these differences are statistically significant when necrosis, ductal dilatation and overall severity of lesions are considered. The results of this study suggest that in the pancreas the toxicity of ethanol is enhanced by interaction with other components of alcoholic beverages. PMID- 7484242 TI - [Primary lymphomas of the brain not related to immunodeficiency]. AB - Primary brain lymphoma (PBL) is a rare disease, the study of which is based on small series and case reports. The AA review new cases of PBL not associated to immunodeficiency at the Portuguese Institute of Oncology of Porto since 1978. Five male patients were found with lymphoma primarily located in the brain, with aggressive histology, IE staging, detected by CAT, NMR or brain scintigraphy and with histologic material collected by stereotaxic biopsy (3 cases) or by craniotomy (2 cases). Three patients were in complete remission (25,5 and 1 year) and 2 were dead with a survival of 12 and 17 months. The cases and literature are discussed. PMID- 7484243 TI - [Invasive lobular carcinoma of the breast with a 10-year evolution]. AB - The authors present a case of an 82-year-old woman with on invasive lobular carcinoma of the breast (ILC) diagnosed ten years ago in a left axillary adenopathy. Physical and mammographic findings appeared as normal. Some considerations are made about the low incidence, the clinical examination and mammographic findings. ILC is reported as being frequently multicentric and bilateral. The uncommon challenge of this clinical case is presented. PMID- 7484244 TI - [A peripheral neuroectodermal tumor of the vulva]. AB - A case of malignant peripheral neuroectodermal tumor occurring in the vulva of a 29-year-old woman is reported. It is a rare malignant tumor with aggressive biological behavior belonging to a wide group, the so-called small round-cell tumors. The patient is well and free of disease 8 months after surgery, chemotherapy and radiation therapy. PMID- 7484241 TI - [The effect of the woman's age, the rate of cleavage and embryo quality on obtaining a pregnancy by in-vitro fertilization]. AB - Multiple factors influence the probability of obtaining a pregnancy through in vitro fertilization (IVF) and embryo transfer (ET). This retrospective study was designed to assess their importance in order to improve prognostic ability and treatment success. 341 consecutive embryo transfer cycles using the same ovarian stimulation protocol were considered and divided in two main groups: 92 cycles in which a clinical pregnancy was achieved and 249 cycles without success. All the embryo transfers were performed in patients from the in vitro fertilization program of the Human Reproductive Unit, Santa Maria Hospital, Lisbon, between January 1991 and December 1993. No significant differences were found between the two groups studied concerning the IVF indications, ovarian response to the stimulation, sperm quality, oocyte maturation and mean number of oocytes retrieved per patient. The women's age was higher in the group which did not achieve a pregnancy, when compared with the pregnant group (p < 0.001), showing a decline of success after the age of 35. Overall oocyte fertilization rate was 88.2% in cycles with pregnancy and 83.5% in cycles without pregnancy (p < 0.02). In the pregnant patients, there was a significantly higher rate of embryo transfers in which all the embryos received had reached at least the four-cell stage at 42-43 hr postinsemination, compared with the non pregnant patients (82% versus 63%, p < 0.001). All the 92 pregnancies originated from transfers of at least one embryo that had undergone two or more mitotic divisions.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7484245 TI - [Systemic lupus erythematosus in the elderly]. AB - The authors report two clinical cases of systemic lupus erythematosus, both in patients over 65 years of age. The discussion includes a brief overview of recent publications on this condition in geriatric patients. Some aspects of systemic lupus in elderly persons are suggested and commented on. PMID- 7484246 TI - [The vertebro-thoracic syndrome]. AB - The authors report a clinical case of outlet syndrome, the only one diagnosed in the outpatient clinic of the Plastic Surgery Department, Santa Maria Hospital, Lisbon, in the last 25 years. The principal etiologic factors, clinical aspects, diagnosis and treatment are reviewed. PMID- 7484248 TI - [Sleep, depression and antidepressive treatments (2)]. AB - Sleep research in depressive disorders has advanced considerably in the last two decades. In this review we highlight the different facets of this research, including neurophysiological descriptions of sleep disturbances in depression and data regarding the effects of antidepressant treatments on sleep. The available information is discussed taking into consideration a possible pathogenic role of certain sleep changes in depression, as assumed by several theoretical models. The scope of these models and their explanatory power are briefly analysed. Some comments are made on conceptual and methodological problems in the field of sleep research in psychiatric disorders. PMID- 7484249 TI - [The reform of medical education. Some reflections]. AB - The author points out the factors responsible for the changes verified in the last decades in the science and practice of medicine and in medical assistance; emphasizes the modern meaning of Medical Education as an evolutive, integrated and coherent process, and points out the scope of the university along with its development; defines the contemporary pedagogical objectives of undergraduate medical education, and points out the strategies which are considered the most appropriate to achieve them; reports the winds of change which have swept medical education in Portugal, and refers the obstacles that restrain its application to practice. Finally, the author emphasizes that the success of any educational reform requires the conscious and motivated engagement of its fundamental agents- the student and the professor. PMID- 7484247 TI - [Auricular fibrillation. Antiarrhythmic treatment revisited]. AB - Atrial fibrillation is one of the most common symptomatic sustained arrhythmias seen in clinical practice. Long-term control of heart rate and maintenance of sinus rhythm often require chronic antiarrhythmic therapy. For patients with disabling symptoms of atrial fibrillation that cannot be controlled with pharmacologic therapy, catheter ablation techniques of the atrioventricular junction and surgical procedures aimed at maintaining sinus rhythm have now been effectively used. The efficacy, risks and limitations of pharmacologic and non pharmacologic therapies are presented in this review article. PMID- 7484250 TI - [Health programs. The management of a program]. AB - An opinion article of health care programmes questioning the effects and benefits of their management. The AA, based on their experience in management/administration, present suggestions to identify, quantify and analyse a problem in health care. After characterising a health care problem, the AA point out the fundamental role of planning (in health care) and invited interdepartmental collaboration between the various levels of health care managers. The eventual mission of health care programmes is stressed, their structural conformity with the external environment (community) and internal environment (health care institutions), the resources affected, their process and the results to be achieved, as well as their co-ordination. The programme manager, with a specific profile and duties, is suggested as an important leadership factor. The conclusion advises health care strategists of the future importance of the programmes. PMID- 7484251 TI - [The right to work and the non-discrimination of individuals infected with the human immunodeficiency virus, HIV/AIDS. The fitness test]. PMID- 7484253 TI - [Hand and foot infections in children]. AB - Hand and Foot anatomic and physiologic characteristics make the infections, located there, acquire specific aspects. Seventy seven in-patients admitted and/or with follow-up at the out-patient clinic of Dona Estefania Hospital with Hand (25) and Foot (52) infections, were reviewed during the period between January 1991 and January 1994. We treated, out-patients with paronychia (7 of the hand and 42 of the foot), one patient with hand pulpitis, and one with dorsum hand cellulitis. The remaining 16 with hand infection (64%) and 15 with foot infection (29%) were being treated with splint in the position of rest, elevation of the affected limb and endovenous antibiotic therapy. In all abscess cases, surgical drainage was conducted, 16 the of hand and 22 of the foot. There were no sequels in hand infection. In foot infection there was one osteitis of the first metatarsus and a cutaneous fistula. PMID- 7484252 TI - [Endobronchial biopsy and cytology. Comparative study]. AB - A retrospective study was performed in order to evaluate the diagnostic accuracy for lung cancer in histological biopsy specimens and in cytological examination taken at fibre optic bronchoscopy. Of 530 patients suspected of having a malignant lesion, 150 had pathological confirmation (group A) and 380 did not (group B). In group A biopsy was diagnosed in 78.7% and cytology in 58% of the cases, of which brushing was positive in 94.1% and washing in only 38.6% of the cases. In group B brushing disclosed the condition of lung epithelium in 91.8% and washing in only 48% of the cases. It is concluded that brushing had a higher diagnostic accuracy than washing in both groups, although inferior to biopsy. PMID- 7484254 TI - [Erythrocytic enzymatic systems and their antioxidative potential in pregnant women undergoing labor induction with intracervical PGE2]. AB - Our purpose is to correlate, the apgar score of the new-borns from induction of labour (IL) with Prostaglandin E2 (PGE2) endocervical gel, and the oxidative environment of the pregnant woman during labour, studying biochemical markers of the erythrocytes. PGE2 is responsible for the regulation of the vascular response in pregnancy, namely for the vasoconstriction caused by Angitensin II. The production of Prostaglandins in vivo depends on mechanisms related to free radicals of oxygen (FRO). The production of FRO is enhanced in normal pregnancy. We have studied two erythrocytic enzymes and plasmatic concentration of PGE2 before and after the IL. Those enzymes are oxireductases-the transmembrane reductase (RTM) and the metahemoglobin reductase (MHR). Their function is to prevent the effects of the FRO on cellular biomolecules namely the endothelium and the red blood cells. This prevention of oxidative stress can facilitate the deformability of the erythrocytes, so these cells can easily transpose the small vessels and bind the oxygen to the tissues. The activity of those enzymes can be modulated by PGE2 used in the IL. We have not found significant variations on the activity of RTM after IL. The activity of MHR was enhanced with statistical significance, 30 minutes after the induction. This enhancement of activity can be a mechanism to prevent the oxidative stress of the induction of labour. PMID- 7484255 TI - [Ret proto-oncogene mutations associated with type 2 multiple endocrine neoplasms (MEN 2). Clinical implications]. AB - Multiple endocrine neoplasia type 2 (MEN 2) is an autosomal dominant cancer syndrome. Clinical features and diagnostic aspects of this syndrome are reviewed with a focus on progress biochemical screening to genetic analysis of those at risk. The clinical implications resulting from the application of the new methods of molecular biology are discussed. PMID- 7484256 TI - [Dysautonomia. An introduction]. AB - Autonomic Nervous System (ANS) dysfunctions may be localized or not, and determine either states of hypo or hyperactivity. This article reviews the structure and function of ANS, methods of evaluation, the main clinical features of its dysfunctions, as well as the most common causes involved and current therapeutic possibilities. Localized dysfunctions and states of hyperactivity are not considered. PMID- 7484257 TI - [Control of breathing]. AB - The control system of breathing can be considered as a closed loop system, consisting of two subsystems: the controlling system and the controlled system. Both systems are defined by their input-output relationships. The controlling system is defined by the Respiratory Centers that are responsible for two separate, but overlapping, patterns: the automatic control pattern and the behavioral or voluntary control. In the controlling system the input is the blood gas value and the output is some parameters of ventilation. The controlled system is characterized by an input of ventilation and an output of blood gas values. In this closed loop system breathing is normally regulated by two anatomically distinct but functionally integrated elements, referred to as the metabolic and behavioral respiratory control systems. The metabolic control is concerned with blood gas homeostasis and the voluntary control relates with activities such as phonation and singing that use the ventilatory apparatus for purposes other than gas exchange. PMID- 7484258 TI - [Teratogenic effects of exogenous agents]. AB - Since the identification of Thalidomide as a potent human teratogen, the possible effects of a large number of drugs, and chemical, physical and environmental agents, on the embryo and the fetus have been raised without support of scientific demonstration. During the last ten years, a large number of papers were published on this issue, making the understanding of teratogenic mechanisms and effects a new field for investigation. This review is to show the recent trends and concepts on human teratogenesis induced by extrinsic agents, making it clear that the majority of them can not be seen as harmful to humans. PMID- 7484259 TI - [Broncho-splenic fistula caused by hydatidosis]. AB - To our knowledge, this is the first case report of a broncho-splenic fistula of hydatid origin. We discuss the clinical, radiological and therapeutic aspects of this rare complication of hydatid disease. PMID- 7484261 TI - [Applications of problem-based learning in the basic medical curriculum]. AB - This article describes a project, which took place between 1992 and 1994, on the application of problem-based learning to three disciplines of the basic medical curriculum: Biochemistry (1st year) Physiology (2nd year) and physiopathology (3rd year). The main objectives of the project were to bring teaching closer to the future professional reality, to stimulate critical thinking and to facilitate independent learning. Despite differences in content and in teaching materials, the trigger event for learning was information given within a clinical context which was further analyzed in small-group tutorial sessions. The project also included the training of the tutors and the development of audiovisual materials. Although no statistically significant differences were found in the final scores between students taught during this period and before, the first group reported a considerable increase in the motivation to learn which, in the case of Biochemistry, was important to change the passive attitude of most students on entering university. The project also helped to create the appropriate atmosphere for the establishment of an office for medical education in the medical school to support other innovations in the field. With this project we wished to contribute, with a concrete experience, to the reform of medical education in Portugal, by demonstrating that problem-based learning can be used in single disciplines without major changes in the basic curriculum. PMID- 7484260 TI - [J. Pinto Correia]. PMID- 7484262 TI - [Medical associations. Tradition and culture]. PMID- 7484263 TI - [To prognosticate, to foretell, to prophesy]. PMID- 7484265 TI - [Lipids in children and young people. A public health problem]. AB - Cardiovascular diseases represent an important Public Health problem among us. It is known that the atherosclerotic process on which they are based is mostly dependent on hyperlipidemia, mainly of those characterized by hypercholesterolemia and starts early in childhood. In this paper some lipid parameters were evaluated in children and youngsters and the prevalence of those at risk was compared with the one found in a population sample of adult people, collected at random, using the same determinations and laboratory methods. The authors conclude that preventive measures should be implemented as early as possible in childhood as there are many children and youngsters with hypercholesterolemia and the latter may become lower; some ideas and recommendations for those measures are pointed out. PMID- 7484264 TI - [Severe poisoning by organophosphate compounds. An analysis of mortality and of the value of serum cholinesterase in monitoring the clinical course]. AB - Ingestion of organophosphate (OP) compounds usually results in severe poisoning. We undertook a retrospective study of 52 consecutive patients admitted with severe OP poisoning to determine the value of serum cholinesterase (SChE) in monitoring clinical course. Considering survivors and non-survivors, we evaluate clinical and laboratory baseline characteristics, severity scores (APACHE II, SAPS II), atropine rate (mg/h), SChE evolution at 24, 72 and 120 h and final SChE (SChE at the day of discharge or death). Mortality in the ICU was 28.9% (n = 15). In both groups SChE showed a trend to increase. In survivors, SChE recovery was statistically significant for SChE 24h-SChE 72 h, SChE 24 h-SChE 120 h and SChE initial-SChE 120 h (p = 0.008, p = 0.00003, p = 0.0002 respectively). In this group a simultaneous decrease in atropine requirements was registered. In non survivors, the rate of atropine remained unchanged up to 120 h. Three groups could be defined in non-survivors according to their final SChE and day of death. Non-survivors-1 (death in the first 24h; 2 patients) and non-survivors-2 (death after the first 24 h; 5 patients) had a final SChE below 10% of normal SChE activity and statistically different from survivors' final SChE. Non-survivors-3 (8 patients) had a final SChE similar to the survivors and death was due to sepsis and multiple organ failure (MOF). We conclude that SChE is useful in OP poisoning diagnosis and also in monitoring clinical course. SChE recovery above 10% of normal seems to correlate with good prognosis. Sepsis and MOF were important determinants of mortality. PMID- 7484266 TI - [The epidemiology of chronic venous insufficiency in Portugal]. AB - The prevalence of chronic venous insufficiency (CVI) was investigated in 44,777 unselected primary care outpatient clinics in 17 of the 20 districts in Portugal, during 1993. The diagnosis of CVI was established clinically by 427 participating general practitioners. CVI was more prevalent in females, with a female to male ratio of 2.1:1. The disease affects all age groups, but in females its prevalence increases sharply between 15 and 20 years, while in males it begins to become important about 10 years later. The maximum age-specific prevalence is reached between 55 and 64 years in both sexes, when CVI is present in 58% of females and in 35% of males. Overall prevalence of CVI was calculated using the data from the population census of 1991, by the direct standardization method. The estimated prevalence of CVI in males is 17.8% and in females is 34.1%, corresponding to 812 thousand and 1,741 thousand cases in Portugal, respectively. In the population over 15 years old, the prevalence is 20.7% in males and 40.8% in females. These estimates have an error of +/- 2.5% with 95% confidence. The geographical distribution of the disease showed different patterns in males and in females. Prevalence decreases from coast to inland in males, and from south to north in females. PMID- 7484267 TI - [The usefulness of bronchofibroscopy in the diagnosis of lung neoplasms in patients with protracted pneumonia]. AB - Fiber-optic bronchoscopy is commonly performed in patients with non resolving pneumonia to exclude endobronchial neoplastic tumours. Radiographic resolution of community-acquired pneumonia is variable, depending on several factors, as the causative agent or host dependent factors. Therefore, the decision to perform a bronchoscopy is sometimes empirical. To describe our experience in this subject we reviewed the clinical registrations and the bronchoscopy protocols of 123 consecutive patients with community-acquired pneumonia that did not show significant radiographic resolution after at least ten days of antibiotic therapy. A histological diagnosis of malignant neoplasia was obtained in 5.6% of the cases. All patients were males, more than 55 years old and heavy smokers (more than 40 pack/year). The comparison between this risk group and the other patients showed statistically significant differences in age and tobacco consumption. Symptom duration, haemoglobin, leukocytes, sedimentation rate, renal or hepatic tests were not significantly different in both groups. We recommend that fiber-optic bronchoscopy must be performed early in heavy smokers, patients over 55 years of age with slow or non resolving pneumonia. In non-smokers or younger patients, it should only be performed after 4 to 8 weeks, unless clinical symptoms justify an earlier observation. PMID- 7484268 TI - [Leg ulcers. The diagnostic and therapeutic options]. AB - Leg ulcers, the cause of great morbidity, are a common malady. Diagnosis is sometimes difficult and inappropriate treatment may lead to serious consequences with the risk of amputation. The authors describe the basis for a simple diagnosis of these conditions as well as the usual medical therapy, with special attention to the local treatment of these lesions. PMID- 7484269 TI - [Muscular diseases in hyperthyroidism]. AB - Hyperthyroidism may present various muscular diseases, namely thyrotoxic chronic myopathy, myasthenia gravis, disthyroid ophthalmopathy and thyrotoxic periodic paralysis. Although infrequent, it is possible to find some of these clinical situations in a medical ward of a general hospital. Differential diagnosis must be made with chronic idiopathic polymyositis and certain congenital myopathies. We describe three patients with hyperthyroidism; one of these patients had chronic thyrotoxic myopathy and another had myasthenia gravis. The third had a thymolipoma and ocular changes probably corresponding to myasthenia gravis. PMID- 7484270 TI - [Cerebral metastasis as the form of presentation of a basaloid carcinoma of the esophagus]. AB - A case of oesophageal basaloid carcinoma is reported. The disease was revealed as a brain metastasis and was found ad initium to be in an advanced stage, with evidence of brain, lung and liver metastasis. The treatment performed was palliative subtotal oesophagectomy followed by roentgen therapy. A year and a half later, the patient is still alive and in home care follow up. The authors end stressing this uncommon presentation for an oesophageal neoplasm and the rarity of its histologic type. They also stress the role of surgery, although palliative, in long survival, very unlike the rapid evolution of the few published cases. PMID- 7484271 TI - [Where intensive care and literature can meet. The conference in homage to Rui Carrington de Costa]. PMID- 7484272 TI - [Helicobacter pylori, nonsteroidal anti-inflammatory agents and gastroduodenal changes]. AB - The author discusses the possible interactions between non-steroidal anti inflammatory drugs (NSAIDs) and Helicobacter pylori (Hp) which may play an important role in the unleashing of gastroduodenal lesions. To our knowledge, AINEs have no influence on the prevalence of infection by Hp and the latter does not seem to influence the development and intensity of the lesions caused by NSAIDs. PMID- 7484273 TI - [The semiology and classification of myoclonias]. AB - Myoclonus can present itself in various distinct clinical contexts. The authors review the possible different types of myoclonus, as a single manifestation or included in a syndrome, based on a semiological and aetiological classification. Special emphasis is given to the group of progressive myoclonus epilepsies. PMID- 7484274 TI - End of the beginning. PMID- 7484275 TI - Reproducibility of DXA absorptiometry: a model for bone loss estimates. AB - The enhanced precision of dual-energy X-ray absorptiometry (DXA) allows the detection of very small changes in bone mineral density (BMD). True clinical changes in BMD in patients must be evaluated with the appropriate error of variance. We evaluated the responsiveness of our measures to true bone loss using a statistical variance components model that characterizes the variability associated with error introduced by the machine, operator, and subjects. Our techniques were applied to data from a prospective study of BMD measurements on spine phantoms and on pre- and postmenopausal women performed on the same day or up to 4 weeks apart with DXA (QDR 1000W, Hologic). Our model determined that most of the error in measurements was introduced by operators' and subjects' variability rather than machine performance. The false-positive rates for true bone change are significantly reduced when the appropriate CV% is used to estimate the significance of bone loss over time. Our study underscores the need to use the appropriate precision error to evaluate the clinical significance of changes in bone mass in individual subjects over time. PMID- 7484276 TI - Effects of resistance training on regional and total bone mineral density in premenopausal women: a randomized prospective study. AB - This study was designed to assess the effects of 18 months of resistance exercise on regional and total bone mineral density (BMD) and soft tissue lean mass (STL) in premenopausal women aged 28-39 randomly assigned to an exercise or control group. Twenty-two exercise and 34 control subjects completed the 18-month training study. All subjects were previously inactive and untrained women. Initial, 5-, 12- and 18-month assessments were made of total and regional BMD and total and regional STL using dual energy X-ray absorptiometry. All subjects consumed a 500 mg/day elemental calcium supplement throughout the study. Initial Ca intake without supplement averaged 1,023 mg/day in total sample. Serum levels of bone osteocalcin and dietary assessments using 12 randomly assigned days of diet records were also completed. Muscular strength was assessed from both 1 repetition maximum (RM) testing of 10 weightlifting exercises and by peak torque for hip abduction/adduction and knee extension/flexion. Training increased strength by 58.1% based on 1 RM testing and by 33.8% based on isokinetic testing at 18 months versus baseline. BMD increased significantly above baseline at the lumbar spine for the exercise group at 5 months (2.8%), 12 months (2.3%), and 18 months (1.9%) as compared with controls. Femur trochanter BMD increased significantly (p < 0.05) in the exercise group at 12 months (1.8%) and 18 months (2.0%) but not at 5 months (0.7%) as compared with controls. No changes in total BMD, arm BMD, or leg BMD were found. There was a 20% increase in BGP in the exercise group as compared with controls at 5 months and this difference was maintained throughout the study. For STL, significant increases for total, arm, and leg were found at 5, 12, and 18 months for the exercise group versus control ranging from 1-6% over baseline. These results support the use of strength training for increasing STL and muscular strength with smaller but significant regional increases in BMD in the premenopausal population. PMID- 7484278 TI - Assessment of spine bone mineral density in ovariectomized rats using DXA. AB - Measurements of lumbar spine (L1-L6) bone mineral density (BMD) and bone mineral content (BMC) of Wistar rats were obtained by dual energy X-ray absorptiometry (DXA) (QDR-1000W, Hologic Inc., Waltham, MA) to estimate reproducibility and investigate age-related changes. In addition we evaluated the accuracy of the technique in female rats. The coefficients of variation (CV) for spine BMD measurements were found to range from 0.73-1.04 in vivo and from 0.36-1.56 in vitro. The in vitro measurements were performed in a 3 cm deep water bath to stimulate an equivalent tissue thickness. Spine BMC, measured in vivo and in vitro correlated closely with the subsequently determined ash weights (r2 = 0.87 and 0.97, respectively). We examined age-related spine BMD by DXA. A relatively constant increase in spine BMD was observed from 6 weeks to 22 weeks; spine BMD remained stable between 22 and 58 weeks. No peak was observed in spine BMD. To evaluate the effect of estrogen deficiency on animals of different ages, we measured spine BMD weekly in female rats subjected to ovariectomy (OVX) or sham operation at 8 and 23 weeks of age. The spine BMDs in each OVX rat were significantly lower than that of the controls. In the 23-week-old rats, bone loss was quite rapid for the first 3 weeks of observation and stable afterward. The BMD of 8-week-old OVX rats increased with body size. We conclude that DXA allows the observation of age-related changes in the spine BMD of rats with great precision.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7484277 TI - Characterization of MCSF-induced proliferation and subsequent osteoclast formation in murine marrow culture. AB - To clarify events involved in 1,25(OH)2D3-stimulated osteoclast-like cell (OCLC) formation in primary murine marrow culture, we have characterized kinetics of precursor proliferation and fusion and their dependence on macrophage colony stimulating factor (MCSF). 3H-thymidine nuclear incorporation in tartrate resistant acid phosphatase positive multinucleated cells (TRAP+ MNCs) was assessed: 3H-thymidine incorporation was greatest when tracer was added during day 4 or 5, with labeled nuclei in 81% (day 4) and 90% (day 5) of the TRAP+ MNCs counted at the end of day 7. The percentage of total nuclei labeled was highest when 3H-thymidine was dosed on day 4 (58%), decreasing to 2% by day 7. Final TRAP+ MNC numbers were depleted by 80% when treated for 24 h with hydroxyurea on either day 3 or 4; this inhibition dropped to 57% and 12% when hydroxyurea was pulsed during days 5 or 6, respectively. The absence of 1,25(OH)2D3 during days 1 4 caused 70% attenuation of TRAP+ MNC formation; however, exposure to 3H thymidine during day 4 in this experiment resulted in subsequent labeling of 81% of the TRAP+ MNCs formed, indicating that precursor proliferation occurred in the absence of 1,25(OH)2D3. To demonstrate that proliferation required MCSF, cultures were exposed to a monoclonal anti-MCSF antibody during days 3, 4, 5, 6, or 7. Inhibition of TRAP+ MNC formation was 85% when antibody was added during day 3. Antibody treatment after day 5 had little effect on the OCLC number. Fusion of precursors showed steady progression with OCLCs containing 4.8 +/- 0.3 nuclei at the end of day 4, 8.3 +/- 0.5 nuclei after day 5, 12.0 +/- 1.3 after day 6, and 13.7 +/- 1.5 at the end of day 7. This steady accretion of nuclei was unaffected by doses of MCSF antibody which blocked proliferation. In conclusion, we have shown that OCLCs arise from an MCSF-dependent expansion of the precursor pool occurring during days 3 and 4. Fusion of these precursors, which begins as proliferation diminishes, is able to progress in the presence of anti-MCSF antibody. These results should help refine the analysis of factors affecting proliferation and fusion of osteoclasts in murine marrow culture. PMID- 7484279 TI - Nitric oxide: a cytokine-induced regulator of bone resorption. AB - Nitric oxide (NO) has been reported to inhibit osteoclastic bone resorption, yet potent stimulators of bone resorption, such as interleukin-1 (IL-1) and tumor necrosis factor (TNF), are known to stimulate NO production. This paradox prompted us to reinvestigate the relationship between NO production and bone resorption in mouse calvarial organ cultures. Control cultures and those stimulated with calciotropic hormones and individual cytokines produced little NO, and under these conditions the NO synthase inhibitor, L-NG-monomethyl arginine (LMMA), had no significant effect on bone resorption. Cytokine combinations were much more potent stimulators of NO production than individual cytokines. Dramatic stimulation of NO production and inhibition of bone resorption resulted when gamma-interferon (IFN) was combined with IL-1 or TNF and these effects were reversed by LMMA. IFN had no effect on bone resorption and little effect on NO production when used alone or in combination with calciotropic hormones, however, suggesting that IFN selectively inhibits cytokine induced bone resorption by generating large amounts of NO. IL-1 and TNF acted together to stimulate NO production but to a lesser degree than when combined with IFN. LMMA inhibited bone resorption induced by IL-1 and TNF, suggesting that lower concentrations of NO stimulate bone resorption. Experiments with the pharmacological NO donor S-nitroso-acetyl-penicillamine (SNAP) supported this view in showing generalized suppression of bone resorption at high SNAP concentrations, but potentiation of IL-1 induced bone resorption at lower SNAP concentrations. We conclude that cytokines are potent inducers of NO in bone and that cytokine-induced NO production has biphasic effects on bone resorption.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7484280 TI - Influence of high glucose on 1,25-dihydroxyvitamin D3-induced effect on human osteoblast-like MG-63 cells. AB - Impaired bone formation due to defective osteoblast function, as reflected in a decreased serum osteocalcin (OC) concentration in the patients with diabetes, has been implicated in the development of diabetic osteopenia. The role of hyperglycemia in this decrease in serum OC concentration was investigated. 1,25 dihydroxyvitamin D3 (1,25[OH]2D3), an active form of vitamin D3, stimulated OC secretion from the human osteosarcoma cell line MG-63 in a dose-dependent manner. Exposure of the cells to high concentrations of glucose for 7 days significantly impaired 1,25(OH)2D3-induced OC secretion as compared with that observed with cells maintained under normal glucose (5.5 mM) or high mannitol conditions. The inhibitory effect of glucose was in a dose-dependent manner up to 55 mM. High glucose (55 mM) also attenuated the 1,25(OH)2D3-induced increase in OC mRNA abundance in MG-63 cells, suggesting that the inhibition of the 1,25(OH)2D3 induced increase in OC secretion by exposure to a high concentration of glucose was, at least in part, mediated at the transcriptional level. High glucose significantly decreased the number of 1,25(OH)2D3 receptors in MG-63 cells, without any change in the dissociation constant for 1,25(OH)2D3; this effect was not mimicked by high mannitol, indicating specificity for glucose. These observations suggest that a high glucose concentration significantly impairs the ability of osteoblastic cells to synthesize OC in response to 1,25(OH)2D3 by reducing 1,25(OH)2D3 receptor number, and that impaired cell function caused by sustained exposure to high glucose contributes to the defect in bone formation observed in the patients with diabetic osteopenia. PMID- 7484281 TI - Growth hormone stimulates bone formation and strength of cortical bone in aged rats. AB - The influence of growth hormone on bone formation, mechanical strength, and composition has been investigated in femur middiaphyseal cortical bone from 2 year-old male rats. The rats were given biosynthetic human growth hormone (bhGH) at 2.7 mg/kg/day in two daily injections for 20, 40, or 80 days, and all animals were killed 80 days after the start of bhGH administration. Control animals were given saline. All animals were labeled with tetracycline on days 41 and 69. Only in the bhGH-80-day group was subperiosteal tetracycline double labeling seen all around the femur diaphysis, and this pattern was found in all animals of the group. Double labeling subperiosteally at the posteromedial aspect was found in all animals of the experiment, but compared with the control group, a 400% and an 800% increase in mineral apposition rate was seen in the bhGH-40-day and bhGH-80 day groups, respectively. Light microscopy and polarization microscopy showed that this newly deposited bone was organized in the same concentric lammellae and had the same direction of the collagen fibers when compared with the surrounding bone formed before the start of bhGH injections. The cortical bone cross sectional area was increased in the bhGH-40-day and bhGH-80-day groups. At the endosteum, scattered labeling was found in animals from all groups, and no differences in medullary cross-sectional areas were seen. The mechanical analysis revealed an increased mechanical strength of the whole diaphyseal bone after bhGH administration. When the data were corrected for dimensions of the diaphyseal bone, no differences in intrinsic mechanical properties of the bone tissue were found. No differences in apparent density of dry defatted bone, ash, and collagen were seen, whereas apparent density of dry defatted bone minus ash was decreased in all groups given bhGH. Correspondingly, a slight increase in ash concentrations of the bhGH-injected animals was seen. bhGH administration also increased the body weight, muscle mass, and total serum IGF-I and thyroxine concentrations. PMID- 7484283 TI - Effects of fluoridated drinking water on bone mass and fractures: the study of osteoporotic fractures. AB - To determine if optimal fluoridation of public water supplies influences bone mass and fractures, we studied 2076 non-black women, all aged > or = 65 years recruited into the Study of Osteoporotic Fractures at the Pittsburgh clinic. Information on fluoride exposure was limited to community water supplies. The variable used in the analysis was years of exposure to fluoridated water in community drinking water supplies. Bone mineral density (BMD) was measured at the spine and hip using dual energy X-ray absorptiometry and at the midpoint and ultradistal radius and calcaneus using single photon absorptiometry. Prevalent and incident vertebral fractures were determined by morphometry. Incident nonspine fractures were ascertained every 4 months and confirmed by radiographic report. Exposure to residential fluoridated water had no effect on bone mass. Women exposed to fluoride for > 20 years had similar axial and appendicular bone mass to women not exposed or women exposed for < or = 20 years. There was some suggestion that women exposed to fluoride for > 20 years had a lower relative risk of nonspine fractures (relative risk, RR, = 0.73; 95% confidence interval [CI] 0.48-1.12), osteoporotic fractures, RR = 0.74 (CI 0.46-1.19), and hip fractures, RR = 0.44 (CI 0.10-1.86), compared with women not exposed, but none of these relative risks was statistically significant. There was no association with wrist or spinal fractures. Our results do not support the findings from recent ecological studies which showed an increased risk of hip fracture among individuals exposed to fluoridated public water. PMID- 7484284 TI - Maxillary molar extraction causes increased bone loss in the mandible of ovariectomized rats. AB - Although osteoporosis is a major public health concern, its effect on oral bone has not been determined. More important may be the effect of estrogen depletion on the response of oral bone to dental treatments such as tooth extraction or pathologic processes such as periodontal disease. Our objective was to determine if maxillary molar extraction increases mandibular bone loss in the ovariectomized compared with a sham-operated control. Fifty-three ovariectomized and 53 sham-operated 6-month-old Sprague-Dawley rats were randomly assigned to the following groups: (1) ovariectomized, adult; (2) sham-operated, adult; (3) ovariectomized, adult, extraction; (4) sham-operated, adult, extraction; (5) ovariectomized, old; (6) sham-operated, old; (7) ovariectomized, old, extraction; and (8) sham-operated, old, extraction. Fourteen days following ovariectomy, the extraction groups had their bilateral maxillary molars extracted. The adult and old rats were sacrificed 114 and 200 days postovariectomy, respectively. The right mandible was tested to failure in three point bending. The bone mineral density (BMD) of the left mandible was measured with high resolution dual energy X-ray absorptiometry. The area fraction and area moment of inertia of mandible sections were determined using image processing software. In the ovariectomized rats, maxillary molar extraction resulted in decreases (p < 0.05) in the failure load (21%), stiffness (39%), BMD (3%), and bone area fraction (8%) of the mandible. However, in the sham-operated rats, these decreases following maxillary molar extraction were less (p < 0.05) than those in the ovariectomized rats and only present in the mandibles of the aged rats.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7484282 TI - The effects of calcium supplementation (milk powder or tablets) and exercise on bone density in postmenopausal women. AB - The etiology of age-related bone loss is unclear but both lack of exercise and dietary calcium deficiency have been implicated in its causation. This 2-year randomized placebo-controlled study was designed to examine the effects of increased dietary calcium and exercise in 168 women who were more than 10 years postmenopausal. The subjects were randomized into one of 4 groups: placebo, milk powder containing 1 g of calcium, calcium tablets 1 g/night, and calcium tablets 1 g/night and an exercise regimen. The exercise group aimed to undertake 4 h of extra weight-bearing exercise per week and were undertaking 10% more activity than other groups at 2 years. Bone mineral density at the lumbar spine, three hip sites, and two sites of the tibia close to the ankle joint were measured at 6 month intervals. Dietary intake was evaluated by a weighed food record, exercise was evaluated by an exercise diary, and blood and urine samples were obtained to examine effects on calcium homeostasis. Individual data points were compared using repeated measures ANOVA and least squares regression. Calcium supplementation by either the calcium tablets or the milk powder resulted in cessation of bone loss at the intertrochanteric hip site (placebo, calcium tablets, calcium and exercise, milk powder -0.81, +0.17, +0.23, and +0.07% per year, respectively; p < 0.05 for all supplementation groups compared with placebo) with similar results at the trochanteric hip site. The calcium and exercise group had less bone loss at the femoral neck site when compared with calcium supplementation alone (placebo, calcium tablets, calcium and exercise, milk powder -0.67, -0.18, +0.28, and -0.18% per year, respectively; p < 0.05 for calcium and exercise compared with calcium alone). There was a significant reduction in the rate of bone loss at the ultradistal site of the tibia (placebo, calcium tablets, calcium and exercise, milk powder -2.5, -1.6, -1.0, and -1.5% per year, respectively; p < 0.05 for all supplementation groups compared with placebo). There was no significant bone loss at the spine site in any group.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7484286 TI - Estimates of volumetric bone density from projectional measurements improve the discriminatory capability of dual X-ray absorptiometry. AB - To determine whether estimates of volumetric bone density from projectional scans of the lumbar spine have weaker associations with height and weight and stronger associations with prevalent vertebral fractures than standard projectional bone mineral density (BMD) and bone mineral content (BMC), we obtained posteroanterior (PA) dual X-ray absorptiometry (DXA), lateral supine DXA (Hologic QDR 2000), and quantitative computed tomography (QCT, GE 9800 scanner) in 260 postmenopausal women enrolled in two trials of treatment for osteoporosis. In 223 women, all vertebral levels, i.e., L2-L4 in the DXA scan and L1-L3 in the QCT scan, could be evaluated. Fifty-five women were diagnosed as having at least one mild fracture (age 67.9 +/- 6.5 years) and 168 women did not have any fractures (age 62.3 +/- 6.9 years). We derived three estimates of "volumetric bone density" from PA DXA (BMAD, BMAD*, and BMD*) and three from paired PA and lateral DXA (WA BMD, WA BMDHol, and eVBMD). While PA BMC and PA BMD were significantly correlated with height (r = 0.49 and r = 0.28) or weight (r = 0.38 and r = 0.37), QCT and the volumetric bone density estimates from paired PA and lateral scans were not (r = 0.083 to r = 0.050). BMAD, BMAD*, and BMD* correlated with weight but not height. The associations with vertebral fracture were stronger for QCT (odds ratio [QR] = 3.17; 95% confidence interval [CI] = 1.90-5.27), eVBMD (OR = 2.87; CI 1.80-4.57), WA BMDHol (OR = 2.86; CI 1.80-4.55) and WA-BMD (OR = 2.77; CI 1.75-4.39) than for BMAD*/BMD* (OR = 2.03; CI 1.32-3.12), BMAD (OR = 1.68; CI 1.14-2.48), lateral BMD (OR = 1.88; CI 1.28-2.77), standard PA BMD (OR = 1.47; CI 1.02-2.13) or PA BMC (OR = 1.22; CI 0.86-1.74). The areas under the receiver operating characteristic (ROC) curves for QCT and all estimates of volumetric BMD were significantly higher compared with standard PA BMD and PA BMC. We conclude that, like QCT, estimates of volumetric bone density from paired PA and lateral scans are unaffected by height and weight and are more strongly associated with vertebral fracture than standard PA BMD or BMC, or estimates of volumetric density that are solely based on PA DXA scans. PMID- 7484285 TI - The effects of intravenous alendronate in Paget's disease of bone. AB - We studied the effects of intravenous alendronate on disease activity in 36 patients with active Paget's disease of bone. Alendronate was administered to 3 groups of 12 patients at doses of 2.5, 5, and 10 mg intravenously daily for 5 consecutive days. The patients were matched for disease activity. Symptomatic improvement was seen in at least 10 patients in each treatment group. Alendronate induced a dose-dependent suppression of biochemical indices of bone turnover in all patients. A significant reduction in the mean fasting urinary excretion of hydroxyproline occurred within 2 days of starting treatment, reaching a nadir at 2-4 weeks, which was most marked in patients receiving 10 mg of alendronate (p < 0.05). There was a slower fall in serum alkaline phosphatase activity with maximal suppression occurring 3 months after the start of treatment. The degree of suppression was least for those receiving 2.5 mg of alendronate (p < 0.05) but no difference in response was observed for the other dosages. The duration of response was also dose-related. A significant fall in the serum calcium and urinary excretion of calcium occurred from the second day of treatment but returned to pretreatment values by 4 months. A transient fall in the mean lymphocyte count was observed, which was similar for each group. This was associated with a short-lived fever in 3 patients receiving 10 mg, in 4 patients receiving 5 mg, and in 2 patients receiving 2.5 mg.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7484287 TI - Validation of accuracy and precision of dual energy X-ray absorptiometry for infants. AB - There is limited information on the validity of bone mineral content measurement by dual energy X-ray absorptiometry (DXA BMC) for use in subjects with low body mass. We evaluated the accuracy and precision of DXA in piglets (body weight 886 5526 g, median 2096 g). Stepwise multiple regression analyses showed that ash weight is the major determinant of DXA BMC (adjusted r2 = 0.98, RMS residual = 3.61 g). The intercept was not significantly different from zero. DXA BMC measurements of other piglets under various clinical situations showed no significant effect from the use of cotton blanket, diaper, or positioning (prone, supine, lateral). In vivo replication of DXA BMC measurements of infants at a postnatal age of from 1-350 days showed a slope of 0.99 and high correlation coefficient (r2 = 0.99, RMS residual = 3.59 g). The intercept was not significantly different from zero, and the average coefficient of variation of duplicate DXA BMC in infants was 2.8%. We conclude that DXA BMC reliably but proportionately underestimates ash weight and is a highly precise method for measuring bone mineral status in young pediatric subjects. PMID- 7484288 TI - Normalization of mineral homeostasis after reversal of osteopetrosis. AB - Whether a radiographic and histologic cure of osteopetrosis includes normalization of mineral homeostasis remains unknown. Thus, we explored the extent of defective mineral metabolism in the microphthalmic (mi/mi) mouse before and after cure. Under basal conditions mi mutants exhibit normocalcemia, hypophosphatemia, and elevated renal 25-hydroxyvitamin D-1-hydroxylase activity. However, administration of PTHrP (3 micrograms/h x 24 h) further stimulated enzyme activity in mi mutants with active disease, to a level no different than that in treated normals. Serum phosphorus levels also declined in mi/mi mice following PTHrP, suggesting a normal renal response to this hormone. In contrast, failure to suppress enzyme function in mi/mi mice following prolonged calcitriol infusion indicates that the observed enhancement of 1,25-dihydroxyvitamin D production occurred secondary to autonomous parathyroid function and/or nonparathyroid hormone-related stimuli. Although an increased fractional excretion and decreased tubular reabsorption of phosphate were demonstrated in mi/mi mice, serum PTH levels were no different in mi mutants compared with normal littermates. Following skeletal cure, the mi/mi mice surprisingly display normal serum phosphorus levels and renal enzyme activity. Moreover, treatment restored normal responsiveness to calcitriol suppression and maintained normal PTHrP responsiveness of enzyme activity. These data indicate that the cure of osteopetrosis in the mi mutant is universal and includes normalization of serum phosphorus and renal 25-hydroxyvitamin D-1-hydroxylase. Furthermore, these data suggest that phosphate depletion of unknown origin is the likely cause of elevated enzyme activity in this murine osteopetrotic mutant. PMID- 7484289 TI - Extracellular matrix stoichiometry in osteoblasts from patients with osteogenesis imperfecta. AB - In previous work, we compared the steady-state levels of specific matrix components in human bone cells derived from patients with osteogenesis imperfecta (OI) to those of age-matched controls. A remarkable finding was the observation that there was a reduction not only in the total levels of collagen, but also in osteonectin and three proteoglycans (a large chondroitin sulfate proteoglycan, biglycan, and decorin). This pattern was observed in patients with and without detectable collagen defects. More recent analysis of extracellular matrix composition have yielded that, compared with age-matched controls, bone cells from OI patients produced higher steady-state levels of fibronectin and thrombospondin. The percentage of these two proteins incorporated into the cell layer pool was also higher in OI than in age-matched controls. In addition, the steady-state levels of hyaluronan and a heparan sulfate proteoglycan were analyzed in both OI and age-matched controls. Although the total (medium + cell layer) steady-state levels of hyaluronan were reduced by 1/3, the percentage of the hyaluronan in the cell layer pool of patients with OI increased between 100 250% of age-matched control. Thus the matrix elaborated by human OI bone cells is not only quantitatively different but also qualitatively distinct from that of age-matched controls. Not only have specific bone cell matrix components (collagen, osteonectin, the large chondroitin sulfate proteoglycan, biglycan, and decorin) been found to be present in reduced levels in OI bone cells, but some matrix components (thrombospondin, fibronectin, and hyaluronan) have also been found to be present in elevated levels in the matrix of OI cells.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7484290 TI - The effects of analytic software and scan analysis technique on the comparison of dual X-ray absorptiometry with dual photon absorptiometry of the hip in the elderly. AB - As part of a longitudinal comparison of bone mineral density (BMD) results originally obtained using a Lunar dual photon absorptiometry (DPA) scanner and later, using a Lunar dual X-ray absorptiometry (DXA) scanner, we compared femur results between DPA and DXA according to DXA analytic software (versions 1.3y and 1.4), and according to the method of placement of the femoral neck box (software algorithm or operator placement according to the appearance of the pair of images) in 58 elderly men and women. The mean BMD at each of three femoral sites was higher using DXA version 1.3y than DPA, but the use of software version 1.4 brought the BMD value closer to that of DPA at all sites. Of 58 scans, 12 (21%) were changed by the operator, resulting in an overall reduction in mean percent BMD difference between scan pairs of 79% (from 1.24% to 0.29%). Although the differences between the DPA/DXA software-driven analysis and the DPA/DXA operator driven analysis appeared small (high r2 values and intra-class correlation coefficients), the increase in sample size that would be required for the same power to detect 2-year changes in BMD if the software-driven analysis was used instead of taking the time to perform the operator-driven analysis was 18%. The findings of this study highlight the need to account for upgrades in analytic software. Furthermore, we present a rational approach for the analysis of serial scans that has face validity and that results in smaller differences between pairs of scans performed on the same individual. The decision to adapt these methods must be based on the relative costs of reducing unwanted scan variability. PMID- 7484292 TI - [Design of a program to prevent adolescent suicide]. AB - In the last decades a notorious increase in suicidal behaviors has been observed in youth. It has become an international health problem which justifies all prevention initiatives that are put forward in some western countries. The authors, given their experience in assessing and treating suicidal youth, design a prevention program addressed to suicidal risk groups, and whose aim is to curb down suicidal rates among adolescents in their Sanitary Area. This program applied in Child and Adolescent Mental Health Center is structured in five levels: Assessing, initial treatment, therapeutical intervention in the medium and long run, other interventions in the community, and evolution. PMID- 7484291 TI - Identification and cloning of human P2U purinoceptor present in osteoclastoma, bone, and osteoblasts. AB - Extracellular ATP acting through purinoceptors may be an important factor in the modulation of bone turnover. In this study we cloned and sequenced the P2U purinoceptor from osteoclastoma, confirming the recently published human sequence. Furthermore, by the reverse transcriptase-linked polymerase chain reaction (RT-PCR) and Southern blotting we demonstrated expression of P2U receptor mRNA in bone, primary cultures of human bone-derived cells, and two osteosarcoma cell lines, Saos2 and Te85. P2U receptor transcripts were identified in alkaline phosphatase-positive human bone-derived cells isolated by flow cytometry providing strong evidence for the expression of the P2U purinoceptor in mature osteoblasts. P2U receptor transcripts were also detected in a purified giant cell population isolated from osteoclastoma, indicating that this receptor is also expressed by osteoclasts. These data suggest that purinergic agonists may play a role in the regulation of bone metabolism. PMID- 7484293 TI - [Development and verification of the Spanish version of the "scanning system" psychiatric interview ("Questionnaires for clinical evaluation in neuropsychiatry"]. AB - The SCAN System (Schedules for Clinical Assessment in Neuropsychiatry) represents the latest stage in a 30 year line of development. It is a set of instruments aimed at assessing, measuring and classifying the psychopathology and behaviour associated with the mayor psychiatric disorders of adult life. The SCAN System has several components: the SCAN interview, the Glossary of differential definitions, the CATEGO5 program and the CAPSE (Computer Assisted PSE version). This latest element is a computerised version of the System, combining the SCAN's interview, Glossary and CATEGO5. The SCAN interview has 3 components, the tenth edition of the Present State Examination (PSE10), the Item Group Checklist (IGC) and the Clinical History Schedule (CHS), whose main features are described in the paper. The CATEGO5 suite of computer programs consists in a set of utilities concerned with data entry and diagnosis. The Entry program is used to enter data recorded manually during a SCAN interview and the diagnosis programs derive diagnosis according to ICD-10 and DSM-III-R/DSM-IV criteria. This report analyses the main methods and results of the "Field Trials" that took place in 20 Centres of different countries. These studies showed that the diagnosis and symptoms scores have a high degree of reliability and that this is also applicable to the spanish version. In conclusion, we consider that the SCAN System is once of the more suitable psychiatric diagnostic instruments currently available. PMID- 7484294 TI - [Prognostic factors in schizophrenic patients]. AB - INTRODUCTION: A series of factors have been identified, linked to the prognosis of the schizophrenia, such as the form of the starting point, the existence of the causing factors, time of evolution, personal and familiar psychiatric antecedents, the civil state, the working situation, the sex, the social class, age of onset and kind of symptomatology. METHODOLOGY: T look into such factors in a sample of 65 schizophrenic patients, it was collected some information about all the variables and relative data in the clinic history, in a formal way, during the admission of the patient in the year 1987/88. In February, 1993, all the cases have been revised and have been qualified as "the best prognosis" or "the worst prognosis", according to some points, such as number of re-admissions, actual symptomatology, level of social and laboural operation. The total sample of patients was dichotomied according to the prognosis variable and the two sub samples were compared in respect to all the variables related to the prognosis. RESULTS: The factors associated with a "best prognosis", include the situation of the laboural activity and the biggest level of adaptation in the previous year to the admission. The presence of self-reference delusion, hallucinations and insomnia, have been associated with a "worst prognosis". CONCLUSIONS: It hasn't been possible to reply, in this study, with the prognosis of the schizophrenia, giving evidence of the methodological difficulties and the heterogeneous results in this kind of studies. PMID- 7484295 TI - [Therapeutic guidelines in chronic fatigue syndrome]. AB - The treatment of CFS is not definitive up till now and it is limited both by ignorance of its causes and by different applicable operative case definitions. It has been etiopathologically related to infectious agents, neuromuscular illnesses, neuro-endocrinous-immunologic alterations and to different psychiatric disorders, particularly depressive disorders. Consequently, a great variety of therapeutic strategies have been tried, most of them with insufficient results. Among the medicamentous ones: immunity activator agents such as recombinant interleukin-2, nonspecific immunitary modulators such as seric gamma globulin, antivirus drugs such as acyclovir, muscular relaxants such as ciclobenzaprine, H2 receptor blockers and steroid and nonsteroid anti-inflammatory drugs such as ibuprofen, naproxen and fulbiprofen. Better results seem to have been obtained with antidepressants, and amfebutamone and serotonin-reuptake selective inhibitors are specially promising. Among the nonmedicamentous strategies, cognitive behavioural treatment can be effective and the so called "psychiatric management of the patient with CFS" has been proposed as a global, pragmatic, individualized, comprehensive approach which must be completed with other interdisciplinary interventions on the patient and his environment. PMID- 7484296 TI - [Assessment of sexuality in patients diagnosed as having an anxiety attack induced disorder (sexuality in patients with anxiety-induced disorder]. AB - Throughout the history of Psychiatry, it is a well known fact the importance of sexuality in Anxiety Disorders. The authors study in a sample of 90 persons, 45 diagnosed as Panic Disorder and 45 controls, several aspects of psychosexuality and its correlationship with anxiety levels, measured by Zung Anxiety Scale and AMDP-AT Scale. The results show that Panic Disorder patients suffer, in a significative way, from a poor psychosexual adjustment, lower levels of satisfaction and higher feelings of guilt in relation to sexuality. There are also differences between Panic Disorder patients with and without agoraphobia. PMID- 7484298 TI - [The madhouse. Considerations on its history and future, 1881]. PMID- 7484299 TI - [Hispanic psychiatry]. PMID- 7484297 TI - [Comorbidity of panic disorder and alcoholism in a sample of 100 alcoholic patients]. AB - Among one hundred patients with alcohol dependence (DSM-III-R) studied in a drug abuse center in the "Bajo Llobregat" area (Barcelona industrial belt it was detected that 27% had life time rate of panic disorder. The age of onset of alcoholism was earlier than the one for panic disorder. In 78.8% of these patients alcoholismo appeared first. 70.4% refer worsening of the panic attacks when drinking large amounts of alcohol. Patients with Panic Disorder: a) are younger (p < 0.05); b) have attended school longer and have higher education (p < 0.01); c) have more alcoholism family history (p < 0.05); d) have more major depressive disorders (0.05) and dysthimic disorder (p < 0.01); e) Worse social functioning according to the GAS (p < 0.01); f) higher score for the Psychological disorders Scale (p < 0.001) and a lower performance at work (p < 0.001) measured by the ASI. The clinical significance of these findings is discussed. PMID- 7484301 TI - [Preparation of a SCID-II-based diagnostic tool for personality disorders. Spanish version. Translation and adaptation]. AB - We carried out the translation and adaptation into Spanish of the original version of the SCID-II (Semistructured Clinical Interview for Diagnosis in Axis II according to DSM-III-R). In the questionnaire we modified the formulation of some items: the questions were changed into assertions and the answer scale was broadened into seven options. In the interview, questions and operative formulations which did not appear in the English version were created. Four trained psychiatrist interviewed in pairs a sample of 60 patients who has previously filled in the questionnaire. The reliability between interviewers oscilates between K = 0.37 for the Schizotypical Personality Disorder and K = 1 for the Avoidant Personality Disorder. The overall agreement (presence or absence of diagnosis) is K = 0.85. With the questionnaire we established 257 diagnoses whereas with the interview we diagnosed 35 cases. PMID- 7484300 TI - [Incidence and multivariable genesis of postmenopausal depression]. AB - The present work tried to find out the postmenopausal depression rates and whether depressive symptoms are due to hormonal disturbances or to multiple variables, not only biological, but also psychological and social. The sample included 128 women with no psychiatric history, pharmacological therapy or recent, important events. Results show a high rate of depression related to different variables, but especially over that of the negative anticipated thinking. PMID- 7484302 TI - [Study of the prevalence of the attention deficit hyperactivity disorder in ten year-old children living in the Valencia metropolitan area]. AB - The point prevalence of Attentional Deficit Disorder with Hyperactivity is estimated in a random sample of 387 10-year-old children selected from the municipal census of the city of Valencia (Spain). A number of associated factors are described, along with the utilization of public health services. Diagnosis was established according to DSM-III-R criteria. The instruments used were: K SADS-E; GAF Scale; Raven Progressive Matrices Test and Sociodemographic, Health and Services Use Information Questionnaire. Prevalence was estimated at 8.01 +/- 2.7% (mild 3.9%; moderate 3.9%; severe 0.3%). The male/female ratio was 2:1. Attentional Deficit Disorder with Hyperactivity was significantly associated with low academic performance and the condition of living with only one parent. These children made use of the psychological support services with significantly greater frequency than non hyperactive children, regardless of the degree of severity or academic performance. PMID- 7484303 TI - [Cannabis and alcohol use as prognosis factors in the short-term progress of schizophrenia]. AB - Data from 62 schizophrenic patients (DSM III), aged 18 to 30, who were treated in the public medical system, and showed a relapse in 1987, after an one-year-follow up, were analyzed. It was observed that cannabis and alcohol use increased the probability of a relapse of schizophrenia in the follow-up period, whilst moderate alcohol use (less than 70 cc of pure alcohol/day) had no influence in the probability of relapse. PMID- 7484304 TI - [The Frankfurt-Pamplona subjective experience scale]. AB - Several instruments have been designed to assess subjective experiences (SE). The Frankfurt Complaint Questionnaire (FCQ) is the most widely used scale to assess SE. This study was aimed to replicate the factor validation of the scale in a Spanish sample. A factor analysis we carried out on the 98-items of FCQ. No a four-factor solution was found, as initially it had been proposed by authors of the scale. A simplified version of the FCQ was derived through procedures of unidimensionality analysis of factors. PMID- 7484305 TI - [Historical study of seven cases of delirium tremens in Spain in the first half of the XIX century]. AB - The authors in the introduction provide an schematic historical scope of reference about the spaniard and european psychiatry from the XVIIIe to the XIXe century. They described the first seven clinical cases of Delirium found by them and published by spaniard physicians in the first half of the XIXe century. They revised mainly the clinical sintomatology, course and evolution, and went into deep detail in the hygienic-dietetical and pharmacological therapeutics used in this period, specially several galenic preparations of opium, tartarus emeticus, cremor, etc. Serapio Escolar y Morales (1808-1874) described the first clinical case of Delirium Tremens in 1839 in a 28 year old man, raising the differential diagnosis between intermittent fever, epilepsy and meningitis, and proposed a useless antiflogistic treatment (bleeding, leeches, poultice) and opium. The second case was described by an anonymous author (J.M.S.Z.) in 1845 in a 42 year old man. Francisco Castellvi y Pallares (1812-1879), in 1845, published another case in a 34 year old male, with an adequate description of the natural history of the disease, healing in 19 days with high doses of watery opium extract. The fourth clinical case was described by R.C.B. in 1846 in a 60 year old man with "alcoholic chorea", "ataxis fever" being the only one with a mortal outcome, treated also with opiates which he did not agree with. The fifth and sixth cases were published by Uliberry in 1847 in two males, 38 and 36 years old respectively. The seventh and last case was described in 1848 by F. Paula Barea, having to highlight the nosological, nosographical and evolutive perspectives in 3 clinical periods and a fourth of convalescence, the treatment differences and the etiopathogenical considerations. Included are 9 tables, 2 graphics and 29 bibliographical references. PMID- 7484306 TI - [Abuse of the monoamine oxidase (MAOI) inhibitors as antidepressive drugs: a critical review]. AB - The cases previously described with tranylcipromine and phenelzine abuse have been reviewed and the main epidemiological, clinical and diagnostic characteristics of them have been analyzed. The aim is to obtain a profile of the patients at risk to develop this kind of problem. In addition, several important questions and the most accepted pharmacological bases of this drugs' addiction are revised. PMID- 7484307 TI - [Father-child hospitalization in the separation anxiety]. AB - Severe forms of separation anxiety disorder with school refusal can be difficult to treat: on the one hand the resistance to outpatient treatment is usual, on the other hand, fear of separation is so overwhelming that hospitalization can be seen as too dramatic. We suggest hospitalizing the child with his or her father as fear of separation with the mother is usually the main problem. We report a case study of a 13 year old child with separation anxiety disorder who refused to go to school and to be separated from his mother. Treatment consisted of hospitalizing the child with his father and to gradually restore his autonomy. The father was given the role of a co-therapist: the main therapeutic decisions about graduated exposure to separation were taken after discussion with the father who passed them on to the child. These structuring interventions re established the father's control which helped to sedate the child's anxiety. PMID- 7484308 TI - The effects of stress and coping on daily alcohol use in women. AB - To evaluate the notion that perceived stress and drinking covary over time, daily logs were periodically collected over a 6-month period from 32 middle-aged women. Results indicated that significantly less alcohol was consumed on high-stress weeks than low-stress weeks. Problem-focused (P-F) coping preference moderated this relationship, with low P-F copers consuming more alcohol per occasion than did high P-F copers but only during low-stress weeks. These findings support a model of stress and alcohol use that includes coping preference as an important moderator of women's drinking. In addition, our data are consistent with the notion that stress can influence alcohol consumption but that low P-F women regulate their use, preferring to delay their drinking until after the termination of the stressor. PMID- 7484309 TI - Norms and sensitivity of the adolescent version of the drug use screening inventory. AB - The distribution and the statistical accuracy of scores on the Drug Use Screening Inventory (DUSI) and the discriminative power of the DUSI for identifying individuals who qualify for a DSM-III-R diagnosis of Psychoactive Substance Use Disorder (PSUD) were examined in 846 adolescents. The subjects with PSUD had higher mean scores, and the distribution of their scores approximated a normal distribution in each of the 10 domains measured by the DUSI. All of the DUSI domains yielded more precise trait estimates for the subjects who had more severe PSUD. Within the normal sample, using a score of 30 on the overall problem density index as the cutoff score, the DUSI correctly classified 95% of the normal sample and 81% of the PSUD cases. PMID- 7484310 TI - Social support for smoking cessation and abstinence: the Lung Health Study. Lung Health Study Research Group. AB - This article evaluates the relationship of social support to smoking cessation and continued abstinence of 3923 men and women with mild to moderate airway obstruction in the Lung Health Study. At both the end of a 12-week group program and after 1 year, men but not women who were supported in quitting were more likely to be successful. Married status facilitated quitting but was less strongly related to long-term abstinence. Participants supported by an ex-smoker who had attended the group program with them were very likely not smoking after 1 year (men, 74.7%; women, 72.4%). Participants supported by a smoker were less than half as likely to have achieved abstinence after 1 year but still had cessation rates greater than 30%. The nature of these relationships has implications for the distinction between women and men in studies of social support and for intervention strategies. Support people should be included in cessation intervention programs. Spouse involvement, however, is more evidently useful for men than for women. PMID- 7484312 TI - Coping capacity in female adolescent substance abusers. AB - Female adolescents who qualified for a DSM-III-R diagnosis of Psychoactive Substance Use Disorder were not distinguishable from depressed and conduct disorder subjects on measures of coping but were differentiable from normal controls. Age of onset of substance use, interval between age of first use and age of diagnosis of abuse, and severity of substance use involvement did not correlate with coping capacity. These findings indicate that deficient coping is not specifically related to drug abuse but rather, where present, is concomitant to comorbid psychopathology among females who have a substance abuse disorder. PMID- 7484311 TI - Contingent reinforcement for reduced carbon monoxide levels in methadone maintenance patients. AB - Five cigarette smoking methadone maintenance patients (MMP) participated in a within-subjects (A-B-A-B) study involving repeated application of carbon monoxide (CO)-contingent payments for reductions in smoke intake. To replicate and extend findings from an earlier study in non-drug-abusing smokers (Stitzer & Bigelow, 1982), we used the same contingent reinforcement procedure that involved the delivery of a monetary payment ($5) for CO readings which were 50% or less than the average value of readings obtained during the prior noncontingency period. Although four of the five subjects earned the contingent payment on at least three of the eight occasions, the overall effect of the intervention on CO level was nonsignificant. Daily smoking records revealed significantly lower rates during the first contingency intervention for the group as a whole. Significant correlations between CO and the time interval since smoking prior to CO measurement were found. The rather modest intervention effects suggest that important parameters be considered when designing contingency management procedures to reduce cigarette smoking in this difficult population of drug abusers. PMID- 7484313 TI - Applying the transtheoretical model to a representative sample of smokers. AB - Interrelationships among key constructs of the Transtheoretical Model are examined for the first time on a large (N = 4,144) representative sample of smokers. The posited relationships between the early Stages of Change (Precontemplation, Contemplation, Preparation) and the Processes of Change, Decisional Balance, and Situational Temptation are generally supported. Precontemplators are found to use the Processes of Change the least and those in Preparation use them the most. Precontemplators also have the least negative attitudes toward their smoking behavior. Precontemplators and Contemplators are tempted to smoke in more situations than those in Preparation. Precontemplators are also the most addicted to smoking, followed by those in Contemplation and Preparation, respectively, when examined on traditional measures of smoking behavior. The implications for smoking cessation efforts are also discussed. PMID- 7484314 TI - Prospective screening of substance dependence: the advantages of directness. AB - This study evaluated the ability of several subtle and direct alcoholism screening scales to identify DSM-III-R-defined substance dependence in a university setting. Study participants were university students (N = 495) who completed the substance use disorder modules of the Diagnostic Interview Schedule along with the Michigan Alcoholism Screening Test (MAST), the Substance Abuse Subtle Screening Inventory (SASSI), and five direct screening questions concerning the consequences of alcohol and drug use. Results indicated that the SASSI had no appreciable ability to identify the 57 study participants who met criteria for a substance use disorder. The MAST also demonstrated limited predictive ability, and results suggested that this is because many MAST items assess the more advanced features of alcoholism, features less common in the substance abuse found among university students. The five direct screening questions were modestly useful, and it is argued that there are advantages to directness when screening for substance use disorders. PMID- 7484315 TI - Differences between responders and nonresponders to cocaine cues in the laboratory. AB - Previous research has shown that a majority of drug-dependent subjects respond to drug cues in the laboratory with increased craving; however, approximately one third are not cue responsive. The current study examined differences between responders and nonresponders to cocaine cues in a sample of 69 cocaine-dependent methadone patients. Subjects completed questionnaires assessing selected dimensions of addiction and participated in a cue-reactivity protocol that assessed both cocaine craving and cocaine aversion in response to cues. Four groups of subjects were identified on the basis of their cue responses: (a) increased craving plus decreased aversion; (b) increasing craving only; (c) decreased aversion only; (d) no increase in craving and no decrease in aversion ("nonresponders"). No group differences were found in severity of cocaine dependence, depression, or on any demographic variable, with the exception of parental status. Group differences were found on measures of cocaine expectancies and self-efficacy. There is a discussion of the bearing of these findings upon explanations offered in the literature for nonreactivity to drug cues. PMID- 7484316 TI - Alcohol expectancies and changes in beer consumption of first-year college students. AB - The present study used a prospective design to evaluate the relationship between alcohol expectancies and the progression of beer consumption of first-year college students over a 2-month period. One hundred and fifty-four first-year undergraduate students completed the Alcohol Expectancy Questionnaire (AEQ; Brown, Goldman, Inn, & Anderson, 1980) and a measure of precollege drinking during their first week of college, and completed a retrospective diary account of alcohol consumption also during the first week and at 1-month and 2-month follow-up. Beer was consumed considerably more frequently than other alcoholic beverages and was used as the dependent measure. The results showed that each AEQ subscale was positively correlated with beer consumption at almost all time points, and the magnitude of these correlations was generally higher for male subjects. Furthermore, the expectancies that alcohol increases social assertiveness and that alcohol is associated with global, positive changes were positively correlated with increases in beer consumption from Session 1 to Session 2 and from Session 1 to Session 3 for male, but not female, subjects. The findings extend previous research by demonstrating that certain alcohol expectancies are related to progressive increases over time in the amount of beer consumed. PMID- 7484317 TI - Reliability of a drug history questionnaire (DHQ). AB - Drug abusers' self-reports are vital to clinical and research endeavors, yet few studies have explicitly examined the test-retest reliability of drug abusers' reports of their pretreatment drug use. The present study evaluated the test retest reliability of drug abusers' reports of (a) lifetime drug use using a drug history questionnaire (DHQ), and (b) demographic and drug-related events. Intraclass and Pearson correlation coefficients revealed reasonably good reliability for most reports of drug use and related events. Further research needs in this area are discussed. PMID- 7484318 TI - The relationship between sexual insecurity, the alcohol expectation for enhanced sexual experience, and consumption patterns. AB - Alcohol expectancies have been shown to correlate with and predict a range of drinking behaviors. This investigation expanded on previous research by examining the differential reinforcement value of alcohol as a means of achieving enhanced sexual pleasure. Results from 303 participants showed that the expectation of enhanced sexual experience was a significant predictor of typical quantity, maximum quantity, and frequency of drinking by sexually secure men. For insecure men and women (both insecure and secure), the expectation that alcohol enhances sexuality was not a significant predictor of drinking patterns. Gender differences are highlighted and the clinical implications for intervention are presented. PMID- 7484319 TI - PTSD substance abuse comorbidity and treatment utilization. AB - The present study investigates the prevalence of posttraumatic stress disorder (PTSD) among a sample of treatment-seeking substance abusers and examines the relationship between PTSD comorbidity and rates of inpatient substance abuse treatment. Eighty-four patients (48 male and 36 female) admitted for detoxification at a private hospital were administered self-report measures of lifetime stressor events, PTSD symptomatology, and prior treatment history. Approximately one quarter of the sample was found to present with significant PTSD symptomatology. Women were more likely than men to have been physically and sexually abused, and women reported experiencing a greater number of traumatic events. Consequently, more women than men were classified as having possible PTSD. With respect to inpatient substance abuse treatment admission rates, the PTSD group reported a greater number of hospitalizations than their non-PTSD counterparts. Implications of these findings for routine trauma screening and more effective treatment for substance abusers with concomitant PTSD are highlighted. PMID- 7484320 TI - Eating disorders in female inpatients with versus without substance use disorders. AB - This study assessed the relationship between substance use disorders and eating disorders in female psychiatric inpatients. Structured diagnostic interviews were reliably administered to a series of inpatients with substance use disorders (n = 67) and a comparison sample without substance use disorders (n = 69). Eating disorder diagnoses as a whole, including eating disorder not otherwise specified, were distributed significantly more frequently among subjects with substance use disorders than among psychiatric controls. Frequencies of the specific diagnoses of bulimia nervosa and anorexia nervosa, however, did not differ significantly between groups. The results suggest that eating disorder features may be overrepresented among female inpatients with substance use disorders. PMID- 7484321 TI - Differences in Asian and white drinking: consumption level, drinking contexts, and expectancies. AB - Despite the heterogeneous nature of interethnic drinking differences within racial groups, research has demonstrated that cautious generalizations can be made when comparing Asian and White drinking patterns. Persons in the United States who identify their race as "Asian" drink much less than their White counterparts, and they report fewer problems. Some have speculated that drinking patterns between racial groups are mediated by culturally influenced social learning. To date there has been very little research examining the expectancy of reinforcement from drinking between different racial, cultural or ethnic groups. The current study of 637 white and 179 Asian university students examined differences in alcohol consumption and problem levels, the social context of drinking, and alcohol expectancies. Results show considerable differences between Asian and White drinking and level of associated problems, as well as similarities and differences in social drinking contexts. It also appears that Asians expected greater tension reduction from drinking despite drinking less overall. Implications for further research are suggested that would link racial, and more specifically, ethnic drinking differences with culturally defined alcohol expectancies. PMID- 7484322 TI - Alternative cut-point scores for the CAST-6. AB - Alternative cut-point scores for the CAST-6, a shortened version of the Children of Alcoholics Screening Test, are examined in three samples: outpatient substance abusers, outpatient psychiatric patients, and medical students. Hit rates for identification are maximized with a lower cut-point for medical students and a higher cut-point for a sample of individuals seeking outpatient treatment for substance abuse. Either cut-point yields equivalent results in an outpatient psychiatric sample. PMID- 7484323 TI - Smoking characteristics of female veterans. AB - Recent studies suggest that the prevalence of smoking among active duty military women is higher than that of active duty military men or civilians of either gender. No data have been published on cessation rates among female veterans. We wanted to report such rates in a group of female veterans at Pettis VAMC Preventive Medicine Clinic and to study predictive factors for smoking cessation and prevention. We identified female patients at Pettis VAMC Preventive Medicine Women's Clinic who had received services in the past 3 years or who enrolled as new patients during the study period; 529 received questionnaires by mail or at the clinic; 409 returned completed surveys. A high proportion (32.5%) of our sample of female veterans were current smokers, and 67% of female veteran smokers reported smoking at least one pack daily. More than one-third of ever-smokers began smoking during military service. Thirty-nine percent of the female veteran sample had CES-D scores of 16 or greater, and current smokers had significantly greater mean CES-D scores than former smokers. Twenty-three percent of current smokers had high nicotine dependence scores (> 7). The highest proportion of CES D scores compatible with depressive symptoms (55%) was seen among current smokers who consumed a pack or more of cigarettes per day. Ever-smokers who screened as depressed were significantly less likely to have quit than those with lower CES-D scores; however, in multivariate models, CES-D scores were not significantly related to cessation. The prevalence of current, ever, and heavy smoking was high among this sample of female veterans attending a VAMC Preventive Medicine clinic. An unexpectedly high prevalence of screening scores compatible with depression was found among this sample, particularly among smokers. This warrants further investigation, particularly in light of the reported relationship between depression and smoking. PMID- 7484324 TI - Two pathways out of drinking problems without professional treatment. AB - This 3-year longitudinal study examined two recovery pathways among 135 problem drinking individuals who never received professional treatment. Almost half (48.3%) of those individuals for whom outcome could be clearly determined became moderate drinkers or stably abstinent. At baseline, individuals who subsequently became abstinent (n = 28) were of low socioeconomic status, had severe drinking problems, and believed their drinking was a very serious problem. Once they began their recovery, they relied heavily on Alcoholics Anonymous as a maintenance factor. In contrast, individuals who became moderate drinkers (n = 29) had higher socioeconomic status and more social support at baseline than did individuals who became abstinent or continued to have drinking problems. A logistic regression on baseline data showed that it was possible to predict which natural recovery pathway an individual drinker would follow. The implications of these findings for alcoholism treatment and policy are discussed. PMID- 7484325 TI - Measuring dependence in smokeless tobacco users. AB - Two scales based on the Fagerstrom Tolerance Questionnaire were developed to measure dependence in smokeless tobacco users. The total score for both scales correlated positively with saliva cotinine levels in subjects, and several individual items produced equally positive correlations. Regression analyses yielded two subsets of three items from each scale that predicted cotinine level. Reports of using smokeless tobacco within 30 min of waking served as a predictor in both models. For the purposes of measuring smokeless tobacco dependence, attention should be given to individual items correlated with saliva cotinine levels that could be used to determine the direction of cessation efforts. Future research should also develop additional items specific to the use of snuff or chewing tobacco and eliminate questions not contributing to the overall scale. PMID- 7484326 TI - Identification of "binge-prone" women: an experimentally and psychometrically validated cluster analysis in a college population. AB - This study investigated the escape model of binge eating through a cluster analysis using standardized measures. A sample of 126 undergraduate women underwent a manipulation of their level of cognition and were asked to "taste test" several flavors of ice cream. Questionnaire data from these women were entered into a cluster analysis. Two groups emerged: women in the "binge-prone" group were significantly more depressed, had lower self-esteem, had more chaotic and extreme eating patterns, and were more self-conscious than those in the control group. In validation work, binge-prone women were shown to report elevated levels of bulimic symptomatology and, when in the presence of a food they enjoyed, to respond to increases in level of cognition by eating more. These results were consistent with some, but not all, of the components of the escape model. PMID- 7484327 TI - Alcohol and tobacco use in the Minnesota Heart Health Program. AB - Correlations between alcohol intake and cigarette smoking have been extensively documented. Prospective data on the relationship between smoking and alcohol remain quite limited. The Minnesota Heart Health Program (MHHP) collected such data as part of a 10-year research and demonstration project intended to reduce the prevalence of heart disease. The study involved six communities, three that received intervention and three, comparison. The current analyses focused upon baseline relationships and longitudinal changes in alcohol and tobacco use. Contrary to prediction, those who quit smoking were no more likely than continuing smokers to reduce alcohol intake. Furthermore, alcohol intake at baseline did not predict smoking status at follow-up. Gender differences were found in a number of comparisons. Further study is needed of changes in smoking and drinking patterns over time in the general population. PMID- 7484328 TI - The correlates of binge eating in two nonpatient samples. AB - Binge eating in two nonpatient samples was examined as a function of actual body weight (expressed as BMI), attempts to control eating, food/weight preoccupation, and neuroticism. All of these factors were univariately associated with binge eating, but multivariate analyses indicated that food/weight preoccupation was the most powerful predictor, accounting for much of the common variance shared by the various correlates. This replicates previous work and suggests that the dynamics of subclinical bingeing are substantially similar to that found in the full DSM-III-R syndrome of bulimia nervosa. PMID- 7484329 TI - Ritalin for nicotine withdrawal: Nesbitt's paradox revisited. AB - This was a unblinded clinical trial of the stimulant methylphenidate (Ritalin) for nicotine withdrawal. Nineteen nicotine-dependent smokers received oral methylphenidate (30 mg target daily dosage) for 5 days following abrupt cessation. Tobacco withdrawal, Spielberger state anxiety, blood pressure, and pulse were measured at baseline, then serially for 7 days. Tobacco withdrawal and state anxiety increased significantly, but 12 (71%) of the 17 smokers who completed the study rated withdrawal relief "very define" and 13 (76%) rated this quit attempt "much easier than other times." Methylphenidate neither increased blood pressure nor blocked cessation-related pulse decrease and appears safe for this indication. Twelve (63%) of the enrolled smokers were confirmed abstinent at day 5. Methylphenidate effect on nicotine withdrawal should be studied in a placebo-controlled clinical trial. PMID- 7484332 TI - Predictors for altering caffeine consumption during stress. AB - The present study investigated whether or not those who self-report increasing use of caffeine when under stress consume caffeine more for the drug (e.g., stimulant) effects. A three-part questionnaire assessing demographic information, caffeine use patterns, and reasons for use was completed by 182 females and 106 males (mean age, 22.4 years; range, 9 to 59). Consistent with earlier research, nearly 50% of the sample reported increasing consumption of coffee or soft drinks when under stress. Hierarchical multiple regression analyses showed drinking for relief reasons as the most consistent predictor for altering caffeine use when under stress. Understanding differences in use patterns will likely help strengthen the ability to describe the extent to which caffeine use contributes to cardiovascular and other disorders. PMID- 7484330 TI - Parental variables associated with bulimia nervosa. AB - The present study examined daughter and parent variables associated with bulimia nervosa, including attitudes about body image, eating and dieting histories, and self-esteem. Subjects were female undergraduates (bulimic, subclinical bulimic, and normal) their parents. Daughters differed in bodymass, with bulimics largest, followed by subclinicals, and finally controls. After daughter's bodymass was covaried, daughters differed in the expected direction on eating and body measures and perceived pressure from mothers to lose weight. Bulimic and subclinical daughters also perceived their mothers, more than their fathers, to pressure them to lose weight. After parents' bodymass was controlled, parents did not differ in eating behavior, dieting and weight history, self-esteem, body importance, or body image (self). Parents also did not differ in their perceptions of their daughters' bodies or the pressure exerted upon their daughters to lose weight after daughter's body-mass was covaried. PMID- 7484333 TI - Prospective correlates of exclusive or combined adolescent use of cigarettes and smokeless tobacco: a replication-extension. AB - This one-year prospective study provides an extension of a previous cross sectional investigation. The previous study found that the number of tobacco products used (i.e., smokeless tobacco, cigarettes), not type of tobacco product, was associated with higher scores on problem-prone variables. In the present study, a sample of 842 southern California seventh-grade adolescents who had not tried either cigarettes or smokeless tobacco were identified and surveyed one year later. Onset of tobacco use was examined as an outcome variable predicted by scores on four psychosocial and two alcohol use variables in seventh grade. Unlike the previous study, females were included in the current study, and the potential moderating effect of gender on the pattern of predictors was examined. Overall, these findings indicate that onset of cigarette smoking or use of both tobacco products is associated with alcohol use, risk taking, and low self esteem. This study provides modest support for the previous investigation. In addition, two of the predictor variables were found to interact with gender. Risk taking was found to have a stronger association with initiation of tobacco use for females than males. Susceptibility to social influence to use tobacco was found to be associated with initiation of tobacco use for males only. PMID- 7484331 TI - Classifying COAs with three variations of the CAST: classification rates, stability, and gender differences. AB - The Children of Alcoholics Screening Test (CAST) was administered to a sample of young adolescents as sixth graders and again one year later. Psychometric properties of the full CAST and a shortened version (CAST-6) were examined, and three different rules for classifying COAs were analyzed. The CAST was found to demonstrate adequate internal consistency and test-retest reliability. When examined as an entire group, the vast majority of respondents maintained the same classification (COA or NCOA) from the first to the second administration. For boys, however, CAST scores and the proportion who were self-identified as COAs declined significantly. Classification rates based on the full CAST and on the shortened version were similar. The use of two confirmatory items, "Ever think your father (mother) was an alcoholic?" however, substantially reduced the percentage of self-identified COAs. Finally, girls were dramatically more likely than boys to endorse items suggesting parental alcoholism, and they were also more likely to self-identify as COAs. PMID- 7484334 TI - Cognitive and situational factors predict alcoholic beverage selection. AB - It has been clearly established that individuals' alcohol consumption tends to covary with the rate and absolute quantity of a drinking partner. Sixty men and 73 women participated in a two-part study designed to examine the role of modeling and alcohol-related expectancies in predicting whether individuals would model the type of beverage selected by a confederate. A stepwise logistic regression designed to predict whether an individual would select an alcoholic or nonalcoholic drink indicated that confederate beverage choice and personal alcohol expectancy (social/physical pleasure) predicted beverage selection. Nonsignificant predictors included gender of the participant, whether participant and confederate were of the same gender, fear of negative evaluation, and typical drinking. The results indicate the importance of evaluating both individual and situational variables in predicting behavior. PMID- 7484336 TI - The effect of type A personality on physiological arousal while playing computer games. AB - Despite the widespread use of computer games, there is a lack of systematic research in the area--particularly on their "addictiveness" potential. Anecdotal evidence suggests that computer game addiction may be due to arousal properties of computer games, although other factors may be important. This study examined the relationship between Type A personality and arousal in computer-game play. It was hypothesised that, during computer-game play, Type A and Type B subjects would have significantly higher heart rates as compared with baseline levels and that Type A subjects would experience a significantly greater increase in arousal when playing a computer game than Type B subjects. These hypotheses were both supported and the results are discussed in relation to arousal, personality, and addiction. PMID- 7484335 TI - Behavioral treatment of cocaine-dependent pregnant women and TB-exposed patients. AB - Health-compromised drug-dependent patients require specialized treatment that addresses both drug use and health risks. This preliminary study examines the efficacy of a contingency management procedure (shaping) on decreasing cocaine use and increasing compliance with the prescribed treatment regimens in two health-compromised cocaine-dependent populations: (i) tuberculin (TB) exposed patients (n = 5) and (ii) pregnant women (n = 7). A multiple-baseline across subjects design was used. There were no contingencies on cocaine use during baseline. During the contingent phase, patients received a monetary reinforcer for (a) successive decreases in the quantity of cocaine and (b) cocaine-free samples. They received a weekly reinforcer if all samples per week met criteria for (a) or (b). During the contingent phase, there was a significant decrease in cocaine metabolite levels and an increase in cocaine-free samples in both populations, with a more robust effect in the TB-exposed group. There was an increase in compliance with prenatal visits among the pregnant women during the contingent phase. Implications for health care are discussed. PMID- 7484338 TI - Gender and family as moderators of the relationship between music video exposure and adolescent sexual permissiveness. AB - This study examined family environment and gender as moderators of an hypothesized relationship between exposure to rock music videos and premarital sexual attitudes and behavior. Results of a survey of 214 adolescents revealed a stronger association between permissive sexual attitudes and behavior and reported exposure to music videos for females than for males. As predicted, the association for females was much stronger for those from unsatisfactory family environments. PMID- 7484337 TI - Psychoactive substance use: some associated characteristics. AB - A questionnaire designed to assess the prevalence of psychoactive substance use and its relation with: (a) central nervous system risk factors, (b) associated disorders (allergies, migraine-type headaches, developmental dyslexia history, smoking, suicide attempt, and sleep disorders), and (c) cognitive-type symptoms, was given to a general population sample of 1,879 university students (mean age = 24.0) from Bogota (Colombia, South America). A prevalence of 3.4% of self reported psychoactive substance use was found. Results indicated that the incidence of some risk factors of minor brain injury or dysfunction, smoking, suicide attempt, and headache was higher among the self-reported psychoactive substance users than among nonusers. Cognitive-type symptoms, except suicide ideation, did not differ between drug users and nonusers. Current results point to a significant association between psychoactive substance use and depressive symptoms, and there was no significant association between psychoactive substance use and psychotic ideation. PMID- 7484340 TI - The changing suicide pattern in Canadian adolescents and youth, compared to their American counterparts. AB - Canada has a high rate of suicide among adolescents and youth--higher than the rate in the United States. The study of variation in societal suicide rates is still guided primarily by Durkheim's (1897) theory which proposed a primarily social integration/regulation theory of suicide. There is evidence that social and economic predictors of suicide vary depending upon the particular subgroup- women or men, and young or old. Rates of birth, divorce, marriage, and unemployment were analyzed and compared to rates of suicide from 1965-1985 in Canada and the United States for particular subgroups. In Canada, measures of domestic integration (divorce and birth rates) and the economy (unemployment rate) predicted youth suicide rates more successfully than they did adult suicide rates. In the United States for the same period, there was less variation in the predictors of suicide by age. Further research as well as caution about overgeneralizing the results are warranted. PMID- 7484339 TI - In search of effective programs to address students' emotional distress and behavioral problems. Part I: Defining the problem. AB - This article discusses the prevalence and seriousness of emotional difficulties and behavioral problems in students, with a special focus on high school students and the obstacles they encounter when addressing these problems. The educational, psychological, and medical literature which addressed the problem over the past five years was evaluated, and 22 references were selected. It was determined that the lack of consensus on the terms used to describe the problem prevented accurate assessment of its prevalence. However, one fifth to one third of the students were found to be affected, leading to serious educational, psychosocial, and economic difficulties--and more are likely to be affected in the 1990s. Since the school and mental health systems have not been entirely successful in addressing the problem, adolescents' patterns of seeking help indicate that peer support groups can be part of the solution. It was concluded that the enormity of the problem requires a low-cost gate-keeping mechanism to facilitate early identification and intervention. Thus, school peer-support programs, if proven effective, may complement traditional mental health services in addressing adolescents' emotional distress and behavioral problems. PMID- 7484341 TI - The relationship of moral reasoning and AIDS knowledge to risky sexual behavior. AB - Two hypotheses were tested in this study: (a) moral reasoning and risky sexual behaviors are inversely correlated; (b) the relationship between AIDS knowledge and sexual behavior is mediated by moral reasoning such that AIDS knowledge and risky sexual behaviors are inversely correlated for higher-level moral reasoners but not for lower-level reasoners. Subjects were 103 undergraduate students who completed an instrument assessing moral reasoning, the Defining Issues Test, and two questionnaires assessing sexual behavior and knowledge about AIDS. Factor analysis identified five sexual risk-taking factors with alphas of .60 or higher. Results supported both hypotheses: (a) Risk taking during sexual intercourse (i.e., lower likelihood of using condoms) and risk taking during anal sex were significantly inversely correlated with moral reasoning; (b) For high moral reasoners, two measures of sexual risk taking were significantly negatively correlated with knowledge about AIDS: as knowledge increased, risk taking during sexual intercourse and engagement in a variety of sexual experiences decreased. The reverse relationship was true for low moral reasoners. Implications for moral development theory and research as well as for AIDS prevention campaigns are discussed. PMID- 7484342 TI - Perceived control and treatment outcome with chronic adolescent offenders. AB - The relationship between perceived control and treatment outcome was explored with chronic adolescent offenders who exhibit internalizing or externalizing behavior problems. Data were examined from the records of 307 male adolescents between the ages of 14 and 18 committed to a state training school for hard-core offenders. Adolescent offenders with externalizing behavior problems, such as aggression, showed a more favorable treatment outcome when they attributed overall successes to their own behavior and when they viewed themselves as worried or anxious. They showed a less favorable treatment outcome when they viewed themselves as generally happy. Adolescent offenders with internalizing behavior problems, such as anxiety or depression, tended to show less favorable treatment outcomes when they viewed themselves as being high in physical competence and when they attributed failures to themselves. PMID- 7484343 TI - Self-perception in late adolescence: an interactive perspective. AB - Self-perception during late adolescence was examined as a correlate of individual temperament. Harter's Self-Perception Scale for College Students and the Adult Form of the Dimensions of Temperament Survey-Revised were administered to 111 college students. It was found that positive self-perception was generally associated with an approaching, flexible, and positive mood pattern, and with high task orientation. Different aspects of self-perception showed distinct relationships with specific temperament dimensions. Self-perception differences for men and women also were noted. An interactive model was used to explain the research outcomes and to suggest environmental features that might be related to the development of positive self-perception. PMID- 7484344 TI - Fathers' and mothers' participation in research. AB - This study investigated the participation rates of fathers, as compared to mothers, in research related to parent and older adolescent psychological symptoms, drinking behavior, and perceived competence. Fathers and mothers did not differ in their rates of participation in the research (59% and 65%, respectively). Additionally, few differences were found between participating and nonparticipating parents. Implications for fathers' and mothers' involvement in research on the family are discussed. PMID- 7484345 TI - Actual and perceived parental social status: effects on adolescent self-concept. AB - This paper proposes a model in which parental social status (PSS) and perceived parental social status (PEPSS) affect adolescent self-concept in distinct ways: PSS affects school achievement and consequently self-concept in the academic domain, whereas PEPSS affects self-concept in the social domain. Two studies of Israeli high school students (N = 569), one on kibbutz and one on urban youth, were carried out. PSS was measured by father's education and occupation, and PEPSS by a new scale. Academic and social self-concepts were tapped by Marsh's SDQ II in the urban sample, and by a combination of the latter and Harter's SPPA in the kibbutz sample. The findings supported the model with one exception: in the urban setting, PSS was not related to academic self-concept. The findings rejected a model underlying the theory of Rosenberg and Pearlin, in which perceived parental social status mediates the effect of parental social status on adolescent self-concept. PMID- 7484346 TI - The Anne Frank Haven in an Israeli Kibbutz. AB - The Anne Frank Haven, founded in 1956, in the Israeli Kibbutz Sasa provides a unique educational program for coping with muticultural and integration problems. It is a holistic, regional junior and senior high school system within the holistic community of three kibbutzim. The Haven has been the subject of much research into "Moral Development," carried out by Wolins (1969, 1971), and mainly by Kohlberg (1971), his doctoral students Reimer and Snarey and other colleagues. In the seventies and eighties they used the Kibbutz example as a model for the "Just Community" approach. In the early nineties, an Israeli group evaluated the success of the program and its rationale, taking into consideration all the "educational factors" of the community, in the Haven, and in the kibbutzim around it. This article offers a comprehensive picture of the Kohlbergian moral developmental research at the Anne Frank Haven, including all the relevant references and evaluations of the Haven as a part of the "Just Community" approach. It concludes with a suggestion for another approach--"Community Education" research in the same Haven--as an example of present and future studies in the area of "Moral" and "Values" education. PMID- 7484347 TI - Cigarette smoking among Korean adolescents: prevalence and correlates. AB - The purpose of this study was to examine the prevalence of cigarette smoking and to explore its relationship with individual, family, school, and psychological factors among Korean adolescents. As part of the Korean Institute for Health and Social Affairs' Adolescent Health Survey, 5,209 eighth graders and 4,677 eleventh graders from 70 junior and senior high schools completed a self-administered questionnaire during school hours in 1989. Current prevalence of cigarette smoking was 8.45% (15.43% of boys and 1.28% of girls). Smoking prevalence increased with age among boys. Based on logistic regression analyses, perceived peer use was most strongly associated with smoking status (OR = 9.97 for boys; and 68.27 for girls). Academic stress, grade, and type of school were also associated with smoking status for both boys and girls. In addition, mother's smoking, birth order, and urbanity were associated with smoking status for boys. Implications of the results for prevention efforts to inhibit the onset of cigarette smoking among Korean adolescents are discussed. PMID- 7484348 TI - Gender and age variations in the self-image of Jamaican adolescents. AB - The purpose of the study was to investigate the relationships among gender, age, and self-image of adolescents attending three secondary schools in Jamaica. The relatively few studies that have been done regarding self-perceptions of these youth are not only dated but have utilized a unidimensional conceptualization of the self. The Offer Self-Image Questionnaire which employs a multidimensional construct of the self was administered to a sample of 174 Jamaican adolescents ranging in age from 14 to 18 years (M = 15.90 years, SD = 1.21). Results revealed statistically significant effects for both gender and age. Gender was found to be significant on one self-image dimension: Morals, while age differences were evident on six dimensions: Social Relationships, Morals, Sexual Attitudes, Mastery of the External World, Vocational and Educational Goals, and Emotional Health. The results in some instances were contrary to those of past research. Discussion focused on cultural socialization and other factors affecting youth in Jamaican society. PMID- 7484350 TI - Impact of the family on the sex lives of adolescents. AB - This article presents the results of three studies as they relate to the influence of the family on adolescents' sex lives. The first study examined sexual and contraceptive practices among girls aged 12-19 in Mexico City. It was found that both communication with the mother and pregnancy history of close female relatives were predictors of initiation of sexual relations, contraceptive use, and pregnancy. The second study examined differences between young men who had gotten their partner pregnant and young men who had not. Again, communication levels with parents were found to influence behavior. The final study examined levels of communication about sex as perceived by parents and their children. Mothers had higher levels of communication with their children about sex than did fathers, and fathers' perception of what they were communicating differed substantially from their children's perception. As a whole, the three studies' results support the hypothesis that the family has a substantial influence on adolescents' sexual and contraceptive behavior and provides justification for the incorporation of such issues as parent-child communication into sex education programs. PMID- 7484351 TI - Menstruation, menarche, and sexuality in the public school curriculum: school nurses' perceptions. AB - Nurses employed by schools and health departments have varying responsibilities for curricula related to menstruation, menarche, and sexuality. Nevertheless, the school nurse is usually a source of information on these subjects whether employed full-time in school health or also participating in other nursing roles in the community. This survey examines the involvement of school nurses in curricula related to human reproduction including contraception. Data about their involvement in these classroom topics provides a basis for evaluating their roles and making recommendations for subsequent continuing education programs for school health nurses. PMID- 7484349 TI - Strategies for coping with sexually transmitted diseases by adolescent females. AB - Coping strategies of adolescent girls in response to the acquisition of a sexually transmitted disease (STD) were evaluated. Perceptions of control over acquisition of an STD and emotional impact were assessed. The sample consisted of 177 adolescent girls recruited from their source of primary care (mean age = 16.8). There were no differences in the use of coping strategies by age group or STD history. Subjects used a wide variety of coping strategies; those who viewed the future acquisition of an STD more negatively used more strategies. Wishful thinking was used by most of the subjects, but was not felt to be helpful; despite this, adolescents with a previous STD were no less likely to use wishful thinking. It was concluded that adolescent health care providers must understand girls' responses to STD and help them develop effective coping skills to reduce the incidence of STD. PMID- 7484352 TI - Perceptions of pregnant/parenting teens: reframing issues for an integrated approach to pregnancy problems. AB - Indisputably, unplanned teen pregnancies have myriad personal, family, and social consequences, but answers to questions surrounding this phenomenon are far less obvious. The purpose of this investigation was to determine correlations, if any, between personal, family, and educational background factors and unplanned teen pregnancy. An anonymous questionnaire was administered to 341 volunteer respondents in 14 Teenage Pregnancy/Parenting Programs (TAPPS). Significant differences among respondents were related to family relationships, race/ethnicity, and sexuality education. These findings provide valuable information for family science researchers, family life educators, health personnel, policymakers, and parents who wish to ameliorate the problems in teen pregnancy/parenting by reframing issues for a more intergrated approach involving both prevention and intervention. PMID- 7484353 TI - Modes of moral judgment among early adolescents. AB - Early adolescence is an important definitional stage during which a value system and behavior code are largely shaped (Levy, 1988). Since one's value system and behavior code govern moral reasoning and resultant behavior, how early adolescents reason about moral/value issues has implications for schools, parents, and society as a whole. This study found that early adolescents use a variety of modes to make moral decisions. These include care--where they wish others not to suffer; justice--where decisions are made according to principles (golden rule), and narrowly concerned or selfish. All three modes were used by males and females; however, males were much more likely to choose the narrowly concerned mode than were females. PMID- 7484354 TI - Factors associated with premature termination of psychotherapy by children. AB - This study reports findings on factors associated with children who terminate psychotherapy prematurely. The sample was composed of 45 children, with an average age of 13.5 years. It was found that children are more likely to remain in treatment if the therapist is perceived as professional and caring, and if the treatment plan is adequately explained. PMID- 7484355 TI - Social skills training for young adolescents: symbolic and behavioral components. AB - The assertiveness program developed by Wise, Bundy, Bundy, and Wise (1991), which focused on the unique problems and peer interactions of young adolescents, was expanded to a 12-week program and presented to a group of 28 sixth graders. Cognitive acquisition and retention of the symbolic information was measured with a pretest, posttest, and follow-up administration of the multiple-choice tests that were designed and used in the prior study. Verbal content of assertive behavior was measured in a pretest and posttest role-play situation. When trained students were compared to a control group on the symbolic measures, a significant effect for treatment (p < or = .000); a significant effect for measures (p < or = .000); and a significant Treatment x Measure interaction (p < or = .000) were found. A comparison of means showed a significant difference between pretest vs. posttest and follow-up (p < or = .000). However, no difference between posttest and follow-up was found (p < .351). Unlike the earlier study, in which girls benefited more from training than did boys, no treatment by gender interaction was found for the cognitive measure (p < .691). Boys and girls benefited similarly in terms of the cognitive acquisition of assertiveness information from this program. However, the results did not show that these students were able to demonstrate assertiveness on the behavioral level. Results are discussed in terms of the difference between simply having stored symbolic information regarding assertive responses and recognizing contexts in which accessing the information would be appropriate. Suggestions for designing programs aimed at bringing about changes in adolescents' assertive behavior that will generalize to their lives beyond the training context are also offered. PMID- 7484357 TI - Factors affecting students' participation in the Cincinnati public schools lunch program. AB - This study reports high school students' attitude toward the school lunch program. Students were randomly selected to participate in a study of factors affecting school lunch participation in the Cincinnati Public Schools (N = 1,804). Using a questionnaire, students were asked their opinions, feelings, and attitudes toward the program. Results showed that the quality and variety of food offered were significant factors affecting the students' decision to participate. The length of the lunch period and the amount of time spent waiting in line also were significant factors. PMID- 7484358 TI - Breakdown of plastics and polymers by microorganisms. AB - The interest in environmental issues is still growing and there are increasing demands to develop materials which do not burden the environment significantly. Awareness of the waste problem and its impact on the environment has awakened new interest in the area of degradable polymers. Biodegradation is necessary for water-soluble or water-miscible polymers because they eventually enter streams which can neither be recycled nor incinerated. It is important to consider the microbial degradation of natural and synthetic polymers in order to understand what is necessary for biodegradation and the mechanisms involved. This requires both biochemical insight and understanding of the interactions between materials and microorganisms. It is now widely requested that polymeric materials come from renewable resources instead of petrochemical sources. The microbial production of polymeric and oligomeric materials is also described. PMID- 7484356 TI - Constructivist developmental theory and therapy: implications for counseling adolescents. AB - Constructivist developmental theory and therapy refers to the integration of developmental theory within the shared context of counseling theory and practice. Drawing on the co-constructivist framework of psychologist Allen E. Ivey, developmental theory and therapy are highlighted within the context of the mental health needs of adolescents experiencing aberrant behaviors and/or significant psychological disturbances. PMID- 7484359 TI - Production of poly(hydroxyalkanoic acid). AB - Poly(hydroxyalkanoic acid) [PHA] is accumulated by numerous microorganisms as an energy reserve material under unbalanced growth conditions in the presence of excess carbon source. In spite of being a good candidate for biodegradable thermoplastics, their high price compared with conventional plastics currently in use has limited their availability in a wide range of applications. With the aim of reducing the high production cost of PHA, much effort is currently being devoted to improve productivity by employing various microorganisms and by developing efficient culture techniques. Several processes recently developed and employed for the production of PHA by various bacteria are described. PMID- 7484360 TI - Horace S. Isbell 1898-1992. PMID- 7484361 TI - Mechanisms in the glucansucrase synthesis of polysaccharides and oligosaccharides from sucrose. PMID- 7484362 TI - Carbon-proton coupling constants in the conformational analysis of sugar molecules. PMID- 7484363 TI - The variable surface glycolipids of mycobacteria: structures, synthesis of epitopes, and biological properties. PMID- 7484364 TI - Nonconventional methods of modification of starch. PMID- 7484365 TI - Carbon-13 nuclear magnetic relaxation and motional behavior of carbohydrate molecules in solution. PMID- 7484366 TI - Site directed mutagenesis to probe for active site components of liver mitochondrial aldehyde dehydrogenase. AB - Mutational analysis allowed us to rule out an essential role for the histidine residues and for serine 74 in mammalian aldehyde dehydrogenase. The later though, was found to be important in coenzyme interaction. The function of the serine could not be replaced by threonine or by cysteine. The absolute requirement for cysteine 302 and for glutamate 268 was verified using mutational analysis. The fact that these two residues are completed conserved among all aldehyde dehydrogenases is consistent with their being essential in the catalytic process. PMID- 7484367 TI - Constitutive and overexpressed human cytosolic class-3 aldehyde dehydrogenases in normal and neoplastic cells/secretions. PMID- 7484369 TI - Tissue-specific expression and preliminary functional analysis of the 5' flanking regions of the human mitochondrial aldehyde dehydrogenase (ALDH2) gene. PMID- 7484368 TI - Metabolism of cyclophosphamide by aldehyde dehydrogenases. PMID- 7484370 TI - Transgenesis of the aldehyde dehydrogenase-2 (ALDH2) locus in a mouse model and in cultured human cells. PMID- 7484371 TI - Class 3 aldehyde dehydrogenase. A northern perspective in the land down under. PMID- 7484372 TI - Studies on the induction of rat class 3 aldehyde dehydrogenase. PMID- 7484373 TI - Mouse class 3 aldehyde dehydrogenases. PMID- 7484374 TI - Cloning and characterization of genes encoding four additional human aldehyde dehydrogenase isozymes. PMID- 7484375 TI - New human aldehyde dehydrogenases. PMID- 7484376 TI - Human class 1 aldehyde dehydrogenase. Expression and site-directed mutagenesis. PMID- 7484377 TI - Retinoic acid synthesizing enzymes in the embryonic and adult vertebrate. PMID- 7484378 TI - Retinoic acid synthesis in the developing spinal cord. PMID- 7484379 TI - Structure and mechanism of aldehyde reductase. PMID- 7484381 TI - Molecular cloning and sequencing of mouse hepatic 11 beta-hydroxysteroid dehydrogenase/carbonyl reductase. A member of the short chain dehydrogenase superfamily. PMID- 7484382 TI - Molecular modelling calculations on the binding of D- and L-xylose to wild-type aldose reductase and its H110Q and H110A mutants. PMID- 7484384 TI - Lysine residues in the coenzyme-binding region of mouse lung carbonyl reductase. PMID- 7484383 TI - Stopped-flow studies of human aldose reductase reveal which enzyme form predominates during steady-state turnover in either reaction direction. PMID- 7484385 TI - Substrate specificity and kinetic mechanism of Tetrahymena 20 alpha hydroxysteroid dehydrogenase. PMID- 7484386 TI - Nitrate esters as inhibitors and substrates of aldehyde dehydrogenase. PMID- 7484380 TI - Expression of human and rat carbonyl reductase in E. coli. Comparison of the recombinant enzymes. PMID- 7484389 TI - The alcohol dehydrogenase system. PMID- 7484387 TI - Purification and characterization of recombinant human placental and rat lens aldose reductases expressed in Escherichia coli. PMID- 7484390 TI - Promoters of the mammalian class III alcohol dehydrogenase genes. PMID- 7484388 TI - Rat and human bile acid binders are members of the monomeric reductase gene family. PMID- 7484392 TI - Molecular evolution of class I alcohol dehydrogenases in primates. Models for gene evolution and comparison of 3' untranslated regions of cDNAS. PMID- 7484391 TI - Class I and class IV alcohol dehydrogenase (retinol dehydrogenase) gene expression in mouse embryos. PMID- 7484393 TI - The role of leucine 116 in determining substrate specificity in human B1 alcohol dehydrogenase. PMID- 7484394 TI - Mutations of human class III alcohol dehydrogenase. PMID- 7484395 TI - Human and rat class IV alcohol dehydrogenases. Correlations of primary structures with enzymatic properties. PMID- 7484397 TI - Purification and properties of murine corneal alcohol dehydrogenase. Evidence for class IV ADH properties. PMID- 7484396 TI - Cloning and expression of a human stomach alcohol dehydrogenase isozyme. PMID- 7484398 TI - Use of a chromophoric reporter group to probe the active site of cytosolic aldehyde dehydrogenase. PMID- 7484399 TI - Mammalian class VI alcohol dehydrogenase. Novel types of the rodent enzymes. PMID- 7484400 TI - Crystallizations of novel forms of alcohol dehydrogenase. PMID- 7484401 TI - Human sorbitol dehydrogenase gene. cDNA sequence and expression. PMID- 7484402 TI - Short-chain dehydrogenases/reductases. PMID- 7484404 TI - Horse liver alcohol dehydrogenase-catalyzed aldehyde oxidation. The sequential oxidation of alcohols to carboxylic acids under NADH recycling conditions. PMID- 7484403 TI - Zinc binding of alcohol and sorbitol dehydrogenases. PMID- 7484405 TI - Analytical approaches to alcohol dehydrogenase structures. PMID- 7484406 TI - Purification and characterization of S-formylglutathione hydrolase from human, rat and fish tissues. PMID- 7484407 TI - Useful mutants of Zymomonas mobilis alcohol dehydrogenase-2 obtained by the use of polymerase chain reaction random mutagenesis. PMID- 7484409 TI - The reduction of propionic anhydride by aldehyde dehydrogenase-NADH mixtures AT pH 7. PMID- 7484408 TI - Studies of the esterase activity of cytosolic aldehyde dehydrogenase using sterically hindered and cyclic substrates. PMID- 7484410 TI - Cloning and characterisation of the cDNA for sheep liver cytosolic aldehyde dehydrogenase. PMID- 7484411 TI - Crystallization of sheep liver cytosolic aldehyde dehydrogenase in a form suitable for high resolution X-ray structural analysis. PMID- 7484412 TI - Progress toward the tertiary structure of (class 3) aldehyde dehydrogenase. PMID- 7484414 TI - Kinetic studies on class 3 aldehyde dehydrogenase from bovine cornea. PMID- 7484413 TI - UDP-glucose dehydrogenase. Structural characteristics. PMID- 7484415 TI - Substrate binding pocket structure of human aldehyde dehydrogenases. A substrate specificity approach. PMID- 7484416 TI - Covalent modification of class 2 and class 3 aldehyde dehydrogenase by 4 hydroxynonenal. PMID- 7484417 TI - The mammalian pineal gland and reproduction. Controversies and strategies for future research. AB - Evidence for a role of the pineal gland and its major hormonal product, melatonin, in mammalian reproduction has accumulated for over three decades. In sorting through the masses of data pertaining to this issue, certain facts are becoming established. For example, the pineal gland is clearly involved in seasonal reproductive cycles via transduction of daylength (more properly nightlength) information in the form of plasma melatonin rhythms. Specific melatonin receptors are found in the hypothalamus and pituitary pars tuberalis of most of the mammalian species examined thus far. Melatonin's mode of action on the reproductive axis is quite variable but may, in many cases, involve modulation of gonadotropin-releasing hormone secretion from the median eminence. Clinical evidence continues to support the idea that melatonin may play a role in the timing of puberty. PMID- 7484418 TI - Ovarian follicular growth, ovulation and atresia. Endocrine, paracrine and autocrine regulation. AB - This chapter describes a finely tuned series of events that results in ovarian morphological changes including follicular growth, dominant follicle selection, oocyte development, ovulation, and corpus luteum formation. These changes are under the obligate control of the pituitary gonadotropins FSH and LH. The signaling mechanisms whereby the gonadotropins stimulate these changes in the ovary are now well described. The gonadotropin membrane receptors have been cloned and the cellular events proceeding from receptor binding to phosphorylation of protein gene regulators, such as CREB, have been elucidated. A whole series of intraovarian paracrine and autocrine regulators have been described as having the capability of "fine tuning" the effects of gonadotropins (see Tables I and II), and they are likely involved in dominant follicle selection and the demise of the corpus luteum. The roles of the individual paracrine regulators are, as of yet, not well known, but the IGF system offers an attractive paradigm for these intraovarian factors. Lastly, we have described the explosion of data on intracellular and intranuclear regulators. The challenge for us will be to understand the physiological significance of the seemingly unlimited number of regulatory possibilities. Much has been learned concerning the regulation of the morphological changes seen in the ovary, but much more remains to be elucidated. PMID- 7484419 TI - Ovulation as a tissue remodelling process. Proteolysis and cumulus expansion. AB - Ovulation, recurring every midcycle of the mammalian female and triggered by a surge of luteinizing hormone (LH) released from the pituitary, is an essential prerequisite for fertilization and subsequent embryonic development. Here we shall describe two of the biological components of the ovulatory response, cumulus expansion (frequently denoted as cumulus maturation) and the rupture of follicular wall, both crucial for the release of a fertilizable ovum. The role of a proteolytic cascade and its regulation by eicosanoids will be emphasized in relation to follicle rupture. The new data implicating cumulus maturation as an essential step for the release of the ovum and the apparent mediatory role of interleukin-1 in this process will be presented. LH/hCG stimulates, in the preovulatory follicles, a cascade of proteolytic enzymes, including plasminogen activator (PA), plasmin and matrix metalloproteinase 1 (MMP-1). These enzymes bring about the degradation of perifollicular matrix and, most notably, the decomposition of the meshwork of collagen fibers which provides the strength to follicular wall. Furthermore, pharmacological blockage of any of these enzymes resulted in inhibition of follicle rupture. LH/hCG stimulates, in addition, an increase in ovarian production of eicosanoids. These include prostaglandins, obtained from arachidonic acid via the cyclooxygenase pathway and leukotrienes, the products of lipoxygenase. Previous studies from our and other laboratories have demonstrated the ability of inhibitors of cyclooxygenase and of lipoxygenases to suppress ovulation in several mammalian species. MK-886, which inhibits the translocation of 5-lipoxygenase (5-LO) from the cytosol and its binding to the membranal 5-LO activating enzyme, suppressed dose-dependently follicular rupture from the treated ovary. Zymographic analysis of ovarian extracts from PMSG/hCG-stimulated rats revealed a band of collagenolytic activity at 52kD, corresponding to human MMP-1 and at 72kD, corresponding to human MMP-2. Both activities were markedly stimulated by administration of hCG and were significantly inhibited by indomethacin, NDGA or MK-886. Thus, eicosanoids seem to mediate LH stimulation of follicular collagenase. Interleukin-1 (IL-1) has been recently implicated in ovulation. The ability of an IL-1 receptor antagonist (ra) to block ovulation in vivo and in vitro has been demonstrated recently. Morphological examination of the ovulatory follicles failing to ovulate suggests that this effect is exerted by inhibiting cumulus oophorus expansion and detachment from mural granulosa cells. In vitro, IL-1ra attenuated the action of hCG and FSH on cumulus expansion and follicular hyaluronic acid synthesis. Thus, IL-1 seems to mediate and/or facilitate gonadotropin action on cumulus expansion, and hence on ovulation. PMID- 7484420 TI - Biochemistry and cell biology of angiotensin-converting enzyme and converting enzyme inhibitors. PMID- 7484422 TI - Angiotensin II receptors. Structural and functional considerations. PMID- 7484421 TI - Neuroendocrine control of reproduction. PMID- 7484423 TI - The angiotensin type 1 and type 2 receptor families. Siblings or cousins? AB - The diverse actions of angiotensin II (AngII) are mediated by cell surface receptors. Molecular cloning techniques have identified two distinct subtypes of AngII receptors referred to as AT1 and AT2. It is now well accepted that multiple forms of the AT1 receptor exist, but similar diversity of the AT2 subtype has not been conclusively demonstrated. Nonetheless, several converging lines of evidence do suggest that multiple AT2 receptors may be present in brain and cultured neuron-like cells lines. For instance, some AT2 receptors are regulated by guanine nucleotides and sulfhydryl-reducing agents, whereas others are insensitive. AT2 receptor populations also exhibit differing pharmacological profiles particularly with respect to their affinity for peptidic and non peptidic ligands. Moreover, a recently developed anti-AT2 polyclonal antisera reveals a unique pattern of immunohistochemical staining in brain and it does not immunoreact with the recently cloned AT2 receptor. Collectively, these results support the hypothesis of multiple AT2 receptors at least within the CNS. Future studies should reveal whether these putative AT2 receptor subtypes result from unique genes or cell-specific post-translational modifications of a single gene product. PMID- 7484424 TI - Angiotensin II in cell growth and matrix production. AB - Glomerular mesangial cells and cardiac fibroblasts have been called "myofibroblasts" because of their phenotypic characteristics (resembling both the fibroblast and muscle cells). Thus, it is not surprising that AII would have similar effects on both cell types, which play critical roles in target organ stress response and wound healing, ultimately leading to remodeling changes. These effects are primarily mediated by the AT1 receptor and include: 1) growth: hyperplasia in cardiac fibroblasts and hypertrophy in normal adult mesangial cells and 2) matrix production: there appears to be an early upregulation of fibronectin message which is later followed by an increase in collagens. It is likely that elevated production of fibronectin may activate signal transduction pathways which lead to increased expression of collagen genes, and which may be critical for the organization and laying down of collagens. Thus, an overall theme that emerges is the impact of AII on both growth and wound repair. Other potential important cellular effects of AII in these systems include: 1) stimulation of growth factors, cytokines, and arachidonic acid products that could have autocrine or paracrine effects, 2) regulation of cell migration and adhesion, 3) alteration of responses to neurohormones, 4) development and maintenance of a differentiated phenotype, and others. Molecular techniques including subtraction hybridization, differential display, antisense knockout, and development of transgenic and embryonic stem cell models will be important in defining the specific role of AII in cardiovascular and renal disease. PMID- 7484426 TI - The adrenal renin/angiotensin system. PMID- 7484425 TI - Angiotensin as a renal growth promoting factor. AB - The kidney has been traditionally considered to be one the pivotal organs involved in the systemic actions of the renin-angiotensin system (RAS) with renin produced in the juxtaglomerular apparatus and angiotensin II (ANG II) as a key player in the regulation of glomerular hemodynamics. However, many studies in the last decade, facilitated by a throughout molecular characterization of all elements of the RAS, have provided convincing evidence that the kidney exhibits a local RAS which may independently function from the systemic actions of the endocrine RAS. Moreover, even local distinct cell populations along the nephron possess all components of a functioning RAS. For example, proximal tubular cells express mRNA and protein for angiotensinogen, renin, and angiotensin converting enzyme (ACE). They bear different types of ANG II receptors with the appropriate signal transduction systems, and these cells also exhibit surface proteases like angiotensinase A which are required for the inactivation of ANG II. Moreover, recent studies in the isolated perfused kidney have clearly shown that proximal tubular cells produce considerable amounts of ANG II and these concentrations exceed approximately hundred times the systemic concentration of the peptide. Besides the well-known regulation of glomerular hemodynamics by contraction of the efferent glomerular arteriole and mesangium cells, ANG II influences transport and acidification processes in proximal and distal tubules. In addition, the octapeptide stimulates metabolic pathways like tubular gluconeogenesis and ammoniagenesis. Accumulating data over the last years derived from in vivo and in vitro studies have demonstrated that ANG II is also a growth factor for renal cells. For example, cell culture experiments have shown that the octapeptide stimulates proliferation or hypertrophy of mesangial cells. In contrast, proliferation of cultured proximal tubular cells is inhibited by ANG II and cellular hypertrophy of these cells is induced. Many studies have provided evidence that early mesangial proliferation/hypertrophy and tubular hypertrophy is a predecessor of the subsequent development of glomerulosclerosis and interstitial fibrosis, situations with irreversible morphological changes of the kidney's architecture leading finally to end-stage renal disease. Therefore, the identification of ANG II as a renal growth factor and a better understanding of its local intrarenal synthesis and growth stimulating effects on different cell types along the nephron may help to develop rational therapeutic interventions to prevent the progression of renal disease. PMID- 7484427 TI - The tissue renin-angiotensin system in the female reproductive tissues. A review. PMID- 7484428 TI - Relevance of the tissue prorenin-renin-angiotensin system to male reproductive physiology. PMID- 7484429 TI - Role of the cardiac renin-angiotensin system in human heart failure. AB - The local effects of angiotensin II (ANG II) on the heart may play an important role for the pathophysiology of cardiovascular disease. Numerous in vitro studies have demonstrated that angiotensin II has distinctive cellular effects in the cardiovascular system which are independent from its effects on blood pressure. These have led to the hypothesis that activation of the angiotensin system in the heart could be of functional relevance for the adaptive processes in several cardiovascular disorders such as cardiac hypertrophy heart failure. This concept has been further supported by clinical studies showing the beneficial effects of angiotensin-converting enzyme inhibitors in these circumstances. In order to study the gene regulation of renin-angiotensin system components in cardiac disorders we investigated the gene expression of angiotensin converting enzyme in human heart failure. Results showed that the enzyme is activated locally in this condition, supporting previous studies in animals. Taken together with recent evidence from genetic studies linking the enzyme to myocardial infarction and cardiac hypertrophy, our findings are in support of the notion that angiotensin converting enzyme plays a central role in cardiovascular physiology and pathophysiology. PMID- 7484430 TI - Transcriptional silencer in intron I of the rat renin gene. AB - In the present study we investigated the influence of intron I of the rat renin gene on the transcriptional activity of its promoter in cell culture. The presence of intron I abolished the transcription of reporter genes (luciferase and lacZ) in the non-renin-expressing human embryonic kidney cell line 293, while it did not significantly affect the activity of the rat renin promoter in rat sceletal myoblast line L8 expressing renin. We conclude from these results that intron I of the rat renin gene contains a tissue-specific silencer element probably also responsible for the transcriptional repression of the endogeneous renin gene in 293 cells. PMID- 7484431 TI - In field-stimulated guinea-pig atria an AT1-receptor mediated increase of noradrenaline release by angiotensin II is seen only in the presence of prejunctional autoinhibition. PMID- 7484432 TI - Association between the renin angiotensin system and anaphylaxis. AB - Patients with hymenoptera venom anaphylaxis before immunotherapy (n = 50) showed significantly lower renin, angiotensinogen, angiotensin I and angiotensin II as compared to healthy non-allergic controls (n = 25) (p < 0.05). A significant inverse correlation between the severity of clinical symptoms and the plasma levels of renin, angiotensinogen, angiotensin I and angiotensin II was found: the lower the levels the more severe the clinical symptoms. Hymenoptera venom allergic patients with repeated anaphylactic reactions during hyposensitizatin did not tolerate the sting of a living insect (n = 6). Sting provocation with a living insect induced clinical symptoms of anaphylaxis in all of the 6 patients. Renin, angiotensinogen, angiotensin I and II remained significantly lower in these patients as compared to healthy non-allergic controls. Likewise, no stimulation of the renin angiotensin system occurred during the anaphylactic reaction. The values of angiotensin II were similar or lower than the values before the anaphylactic reaction. In contrast, patients with successful immunotherapy (n = 27) who tolerated the sting of a living insect, renin, angiotensin I and II were significantly higher than in patients without immunotherapy. After immunotherapy, the values for renin, ANG I and ANG II were similar to the values found in healthy non-allergic controls. Patients with a history of hymenoptera venom anaphylaxis (n = 22) showed significantly lower ANG II concentrations in their leukocytes as compared to healthy non-allergic controls (n = 24). Successful immunotherapy induced a significant 9-fold increase in ANG II as compared to patients without immunotherapy. These findings suggest a possible role of the renin angiotensin system in hymenoptera venom anaphylaxis. PMID- 7484433 TI - Cloning, expression and regulation of angiotensin II receptors. AB - Complementary DNAs for angiotensin II type 1 receptor isoforms AT1A and AT1B were cloned by expression cloning from bovine adrenal and rat vascular smooth muscles. Human AT1 receptor was also cloned. Seven transmembrane structures emerged. The AT1 type receptor interacted with more than one type of G-proteins. The ligand binding site of AT1 involving Arg167, Lys199, and Asp263 has been identified by site directed mutagenesis. The regulation of the receptors occur at many stages. The isoform, AT2, was also expression cloned from rat pheochromocytoma cells. Although its ligand binding is not affected by stable GTP analogs, it is a seven transmembrane domain receptor. It mediates the modulations of phosphotyrosine phosphatase by angiotensin II and AT2 specific CGP42112A. The modulation was abolished by pertussis toxin. Thus, AT2 belongs to a new class of angiotensin receptors with unique signalling and regulatory mechanisms. AT1 mediates cellular growth. Interestingly, AT2 expression is inversely related to the mitogenic activity of cells. PMID- 7484434 TI - The relationship between the adrenal tissue renin-angiotensin system, internalization of the type I angiotensin II receptor (AT1) and angiotensin II function in the rat adrenal zona glomerulosa cell. AB - Many data suggest that the elements of the tissue renin-angiotensin system (RAS) in the adrenal cortex are mostly located in the zona glomerulosa. The relationship of this paracrine/autocrine system with the cellular localization of the angiotensin II (AII) receptor has not bee clarified. Using a specific monoclonal antibody (6313/G2) to the first extracellular domain of the type 1 receptor (AT1), we show here that most of the receptor is internalized in the rat glomerulosa cell. This may result from tonic stimulation by the tissue RAS, and consequent permanent receptor occupancy. When viable glomerulosa cells are incubated with 6313/G2, the receptor is transiently concentrated on the cell surface, and aldosterone output is stimulated. This stimulated output is enhanced by neither threshold nor maximal stimulatory concentrations of AII amide, although the antibody does not inhibit AII binding to the receptor. The antibody directly stimulates inositol trisphosphate (IP3) generation, but, while having no intrinsic action on protein kinase C (PKC) activation, significantly inhibits the PKC response to angiotensin II. The data suggest that although the receptor is mostly internalized, recycling to the plasma membrane is constitutive, or regulated by unknown factors. Retention of the AT1 receptor in the membrane is alone enough to allow sufficient G protein interaction to generate maximal steroidogenic effects, through IP3 generation. PKC activation induced by angiotensin II has no bearing on steroidogenesis in the dispersed glomerulosa cell system. PMID- 7484435 TI - Endocrine, paracrine and autocrine regulation of testicular steroidogenesis. AB - Testicular steroidogenesis takes place almost exclusively in Leydig cells. Some metabolism of the androgens produced by Leydig cells takes place in seminiferous tubules, especially in the immature animal (e.g. aromatization and 5 alpha- reduction). Luteinizing hormone (LH) is the main tropic regulator of Leydig cell function, without which quantitatively important androgen production is not possible. LH acts through a receptor that belongs to the seven times cell membrane spanning, G protein associated, receptor family, and cyclic AMP is the main second messenger of its signal transduction. Information about the involvement of other signal transduction systems in LH action has also emerged recently. The action of LH is under manyfold modulation by other hormones (e.g. prolactin, growth hormone and insulin), growth factors and bioactive peptides. In this modulation, various paracrine and autocrine mechanisms play an important role. Seminiferous tubules influence the development and function of adjacent Leydig cells through several growth factors. When germ cells are damaged, Leydig cells in the vicinity proliferate faster. Leydig cell morphology also depends on the germ cell composition in the neighbouring seminiferous tubules, and certain stages of the seminiferous epithelial cycle increase the Leydig cell capacity to produce testosterone. Also negative modulation of Leydig cells by Sertoli/germinal cell derived factors has been demonstrated. However, the physiological importance of the paracrine and modulatory influences of the different hormones and growth factors still remains obscure since almost all information has so far been obtained from in vitro studies. In the study of testicular steroidogenesis, the main switch of the function, LH action, is well known whereas the role of the "in house" circuits of paracrine and autocrine regulation remain to be elucidated. PMID- 7484437 TI - Angiotensin peptides in the brain. PMID- 7484436 TI - AT1 receptors and angiotensin actions in the brain and neuronal cultures of normotensive and hypertensive rats. PMID- 7484438 TI - Brain angiotensin and the female reproductive cycle. AB - The results consistently show from experiment to experiment that there is a surge of brain Ang II prior to the well known preovulatory LH surge. It should be pointed out that these experiments have been carried out by two different laboratories and with the help of different experimenters and some of the experiments have been repeated. Therefore, the consistency of the results is reassuring. It does appear that Ang II increases in the brain, specifically in the hypothalamus, probably in cells of the paraventricular nucleus about 1 hour before the LH levels in plasma rise to a peak. Since LH release from the anterior pituitary gland is stimulated by the release of LHRH from the arcuate nucleus into the median eminence, the results would suggest that Ang II stimulates the release of LHRH. The peak in the OVX of Ang II treated rats is sharp and short lasting with a second, later peak. The LH surge follows the first peak and a second rise in LH follows the second Ang II peak. These data suggest that brain Ang II synthesized and stored in the brain plays a critical role in the female reproductive cycle by initiating the LH surge. The regulation of Ang II may be by estrogen and progesterone, but as the increase in angiotensinogen mRNA was not marked, the surge of Ang II appears to result more from the sudden release of stored Ang II than its synthesis. Thus, the question is what releases Ang II. Earlier studies showed that catecholamines release Ang II from neurons and not from glia involving alpha 2 receptor blockade to increase norepinephrine by inhibiting reuptake (7). An interaction between catecholamines, Ang II and LH had also been suggested earlier (18, 19). Therefore, a series of events triggered by steroids in proestrus may begin with increases in norepinephrine activating neuronal alpha 2 receptors and precipitating release of brain Ang II. This is represented diagrammatically in Figure 15. The Ang II surge stimulates the cells containing GnRH (gonadotropin releasing hormone) in the arcuate nucleus. The effect of Ang II on multiple GnRH cells amplifies the effect and GnRH is released into the portal vessels of the pituitary to stimulate the large LH release, from gonadotrope cells in the anterior pituitary, into the plasma that produces the LH surge. The effect of the LH surge is ovulation which ends the estrogen build up.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 7484440 TI - The ovarian renin-angiotensin system. A paracrine-intracrine regulator of ovarian function. AB - We present a review that considers the regulation of ovarian function by the ovarian renin-angiotensin system. Evidence is presented that shows the ovarian renin-angiotensin system can regulate normal ovarian function by paracrine and intracrine mechanisms. In addition, the complexity that is inherent in the renin angiotensin system conferred by multiple biological angiotensin peptides and multiple receptors is discussed. Evidence that a dysfunctional OVRAS induces ovarian pathology such as polycystic ovarian disease is also presented. PMID- 7484439 TI - Is estrogen-induced pituitary hyperplasia and hyperprolactinaemia mediated by angiotensin II? PMID- 7484441 TI - The pathophysiology of ovarian hyperstimulation syndrome (OHSS). A proposal role of the ovarian derived prorenin to angiotensin cascade (ODPAC). PMID- 7484442 TI - Paracrine regulation of the bovine ovarian prorenin-renin-angiotensin-system. AB - In the bovine ovary, prorenin production by theca cells is known to be regulated by LH. In the present study the aim was to evaluate whether LH-stimulated prorenin production could be further modulated by intraovarian factors in vitro. Theca cells were isolated from bovine ovaries by enzymatic dispersion, purified over Percoll gradient and cultured under serum-free conditions with LH/8Br-cAMP in the absence or presence of different steroids and growth factors and the amount of prorenin secreted into the medium was measured. None of the steroids used (androstendione, estradiol, progesterone) influenced the basal or LH stimulated prorenin production. In contrast, cytokines and growth factors, like TNF alpha, TGF alpha, TGF beta and bFGF proved to be important regulators of prorenin synthesis. Whereas TNF alpha, TGF alpha and bFGF significantly reduced the LH- and 8Br-cAMP-induced prorenin synthesis at a site distal to cAMP formation, addition of TGF beta led to a further increase in the amount of prorenin secreted into the medium. None of the agonists had an influence on prorenin production by itself. The observed effects of cytokines and growth factors seemed to be confined to prorenin production only, since cell number, cell viability and steroidogenic response were not at all influenced by the agonists. We conclude that, although LH appears to be the primary regulator of ovarian prorenin production, several paracrine/autocrine intraovarian factors may be involved in "finely tuning" the secretion of prorenin, which is necessary for maintaining the differentiated state of the follicle. PMID- 7484443 TI - Regulation of angiotensin II receptor expression in ovarian follicles. A review. AB - The regulation of angiotensin (Ang) II receptor expression in ovarian follicles was reviewed. Ang II receptor expression, assessed as the Ang II binding capacity, varied with the animal species and the developmental stage of the follicle. In cows, the expression correlated positively with follicular size and negatively with the active renin concentration in the follicular wall tissue. No relations were found to the follicular fluid concentrations of prorenin, estradiol or progesterone. In cell cultures, luteinizing hormone up-regulated Ang II receptor expression in cows, whereas it was decreased by follicle-stimulating hormone, Ang II and testosterone in rats. These data support the concept of an active and regulated RAS in ovarian follicles. The species differences in Ang II expression suggest varying functional roles of the RAS between species. PMID- 7484444 TI - Regulation of utero-placental prorenin. AB - Prorenin (Pro) is synthesized in a number of human utero-placental tissues, including chorion, decidua, villous placenta and probably mesenchymal cells. The release of Pro from these extra-renal tissues follows new protein synthesis and appears to utilize the constitutive secretory pathway. Unlike processing in the kidney, very little of the Pro is subsequently cleaved to the smaller product (active renin). Primary signals which regulate Pro include protein hormones and peptides (relaxin, endothelin, hCG), amines (epinephrine, norepinephrine, and related beta adrenergic agents), and eicosanoids. These agents increase the mRNA for prorenin at a time before peak secretory effects are noted. Other extracellular signals have negative regulatory effects. These include angiotensin, endotoxin and cytokines (TNF-alpha and interleukin-1 B). There is also evidence that glucocorticoid receptor activation has an inhibitory effects on Pro release in placenta. Second messengers involved in the regulation of Pro include cyclic AMP and protein kinase A (PKA), protein kinase C (PKC), and calcium. The possible biological effect(s) of the extracellular Pro are unknown but may be due to direct generation of angiotensin I. Since angiotensin-peptides have a number of trophic effect on both vascular and non-vascular tissues, regulation of utero-placental Pro by autocrine, paracrine or endocrine signalling may be critical in normal fetal and/or placental development. PMID- 7484445 TI - The choriodecidual renin controversy revisited. PMID- 7484446 TI - Coda. Tissue renin-angiotensin systems, 1994. PMID- 7484447 TI - Gonadotrophin control of testicular germ cell development. AB - Successful and complete male germ cell development is dependent on the balanced, endocrine interplay of the hypothalamus, the pituitary and the testis. The hypothalamus secretes gonadotrophin-releasing hormone in a pulsatile manner which, in turn, elicits the pulsatile release of the gonadotrophins LH and FSH from the pituitary. Luteinizing hormone stimulates spermatogenesis indirectly via testosterone, whereas FSH acts directly on the seminiferous tubules. The synthesis and release of gonadotrophic hormones is under the feedback control of testosterone. Whether other testicular peptides such as inhibin and activin are also involved is not yet clear. Luteinizing hormone/testosterone and FSH are the prime regulators of germ cell development. On their own, these hormones are capable of exerting clear-cut stimulatory effects on the spermatogenic process. However, the quantitative production of spermatozoa generally requires the presence of both LH/testosterone and FSH. Since receptors for androgens and FSH are confined to the somatic cells of the testis, the trophic effects of these hormones on germ cells must be indirect. However, it is not known as yet precisely which genes/factors mediate the beneficial effects of androgens and FSH on spermatogenesis. The gonadotrophic hormones have been found in a number of isoforms and multiple transcripts of the LH and FSH receptor have been detected. Therefore, the possibility must be considered that certain forms of male infertility could be due to dysfunctional hormones and/or mutated receptors. PMID- 7484448 TI - Current aspects of autocrine and paracrine regulation of spermatogenesis. PMID- 7484449 TI - Role of the epididymis in mediating changes in the male gamete during maturation. AB - This article reviews recent knowledge about events occurring in the epididymis that are important for sperm to fertilise eggs. Well established concepts are stated without references (see Cooper, 1986 for older literature) but recent references are included where they throw light on mechanisms of epididymal function. During their sojourn in the epididymis spermatozoa acquire the capacity to move and to fertilise eggs; they are then stored in a quiescent state prior to ejaculation. The ability of sperm to undergo the events of fertilisation are developed as a result of interactions with certain epididymal secretions. Increases in our knowledge about the genes coding for epididymal secretions has not yet been matched by similar insight into the role that these secretions play in the maturation process. However, information about the changes that occur to the sperm cells during maturation permit certain scenarios to be sketched that may reflect reality. This review is one such attempt to bring the epididymal sperm-epithelial secretion into focus. PMID- 7484450 TI - Mutational analysis of 16S ribosomal RNA structure and function in Escherichia coli. PMID- 7484452 TI - Thrombophilia: the discovery of activated protein C resistance. PMID- 7484454 TI - Menkes and Wilson diseases. PMID- 7484451 TI - The origin of numerical chromosome abnormalities. PMID- 7484455 TI - PAX and HOX in neoplasia. PMID- 7484453 TI - Dystrophin, its gene, and the dystrophinopathies. PMID- 7484456 TI - Sex determination and its pathology in man. PMID- 7484457 TI - Meiotic sister chromatid recombination. PMID- 7484458 TI - Mapping of mammalian genomes with radiation (Goss and Harris) hybrids. PMID- 7484459 TI - The CD1 family: a third lineage of antigen-presenting molecules. PMID- 7484460 TI - Molecular and cellular aspects of X-linked agammaglobulinemia. PMID- 7484461 TI - The common gamma-chain for multiple cytokine receptors. PMID- 7484462 TI - Self-tolerance checkpoints in B lymphocyte development. PMID- 7484463 TI - The regulation of pulmonary immunity. AB - No evidence has emerged which suggests that the principles of immunity derived from studies on cells from other body sites are contradicted in the lung and its associated lymphoid tissue. What is clear, however, is that the environment dictates the types of cells, their relationship to one another, and what perturbing events will set in motion either the development of an "active" immune response or tolerance. Investigating mechanisms for the development of lung immunity has increased our understanding of how human diseases develop and is continuing to suggest new ways to manipulate pulmonary immune responses. Demonstration that lung cells regulate both nonspecific inflammation and immunity through the expression of adhesion molecules and the secretion of cytokines offers hope for ways to design more effective vaccines, enhance microbial clearance in immunosuppressed hosts, and to suppress manifestations of immunologically mediated lung disease. Important lung diseases targeted for intensive research efforts in the immediate future are tuberculosis, asthma, and fibrotic lung disease. Perhaps even the common cold might be conquered. Considering the pace of current research on lung immunity, it may not be too ambitious to predict that these diseases may be conquered in the next decade. PMID- 7484464 TI - Positive selection of thymocytes. PMID- 7484465 TI - Rare, new and emerging helminth zoonoses. PMID- 7484466 TI - Human lice and their management. PMID- 7484468 TI - Population genetics of parasitic protozoa and other microorganisms. PMID- 7484467 TI - Ticks and Lyme disease. PMID- 7484469 TI - A "bicycle-handlebar" type of incision for primary and secondary abdominoplasty. AB - The authors register a revision of the literature regarding the types of low transversal pubic incisions used in abdominoplasty. Since 1987, the traditional open W-type incision changed to a new type called the bicycle handlebar. In this procedure, the pubic segment remains in a lower line at the level of the pubic hair, while the lateral limbs are in a high position. The anterior superior iliac spine is used as reference. Different types of procedures where the umbilical skin hole reaches the pubic incision are described and illustrated. This procedure is also used in secondary abdominoplasty. In all cases, the incision's new position should remain hidden under a bathing suit. PMID- 7484470 TI - Tumescent and syringe liposculpture: a logical partnership. AB - Liposuction has been traditionally performed under general anesthesia. Standard instrumentation for the procedure has included blunt-tipped suction cannulae connected to an electric vacuum pump by noncollapsible tubing. A subcutaneous injection of Lidocaine with Epinephrine is routinely employed to minimize blood loss during the procedure. This infiltration has been described as the "wet technique," but it is not a method to supplant general anesthesia. The tumescent technique, a method of infusing very large volumes of dilute lidocaine with epinephrine solutions, has been advocated as a satisfactory means for providing conscious anesthesia for liposuction procedures, avoiding the need for general anesthesia. The syringe technique employs blunt-tipped suction cannulae connected to a syringe. Drawing back the syringe plunger generates the negative pressures needed to remove fat during liposuction and replaces the electric vacuum pump and connecting tubing traditionally used for this procedure. This study evaluates the combined tumescent and syringe techniques for liposuction. One hundred consecutive patients were treated with the tumescent technique as the sole means of anesthesia and the syringe technique as the sole means of performing liposuction. A modified tumescent formula is presented. A comparison of liposuction aspirates using this modified tumescent technique is compared and contrasted to liposuction aspirates obtained using the "dry technique" and the "wet technique." A historical review of the syringe technique and its perceived attributes is also presented. Technical descriptions of the tumescent infusion method, tumescent fluid formulation, and suggested patient sedation and monitoring is presented. Photographic documentation of patients who underwent the combined tumescent and syringe liposculpture treating various body areas is shown. A critical analysis of the limitations of this combined technique is also described noting added time requirements, difficulties with under-correction of deformities, and need for reoperation, methods for determining the "end-point" for the procedure, as well as addressing large-volume liposuction problems. The conclusion reached by this study is that combining the tumescent technique and the syringe technique is a logical partnership. Each method complements the other, allowing liposuction to be performed with considerable advantage over traditional methods. These advantages include eliminating the need for general anesthesia, lessening blood loss and postoperative bruising, greater accuracy, precision, and overall high patient satisfaction. PMID- 7484472 TI - The saddle lift for tight thighs. AB - Efforts to tighten the lateral thigh skin and subcutaneous tissue after liposuction may result in conspicuous scars on the hips, groin crease, or medial thighs. We have developed a technique that enables the excision of skin and subcutaneous tissue, measuring about 20 x 10 cm2, from the inner thigh and central buttocks crease where the scar can be completely hidden. We present a patient who, after appropriate liposuction, had a nearly perfect silhouette but who continually complained about the looseness of the skin on her lateral and posterior hips. The patient suggested that we remove the skin and subcutaneous tissue from the middle portion. We did so and the results were surprisingly good. We recommend this procedure for those patients in whom no external scar is acceptable and who require tightening of the skin of the upper thighs and buttocks. PMID- 7484473 TI - Maliniac Lecture. Plastic surgery of the breast: a woman's decision, a surgeon's challenge. PMID- 7484474 TI - Endoscopic plastic surgery: the endoscopic evaluation of implants after breast augmentation. AB - There is an ongoing discussion about the possible connection of gel-filled silicone implants for breast augmentation and the development of connective tissue disease. In the absence of any proven link and irrespective of the difficulty to prove such a causality, these discussions lead to great concern for the hundreds of thousands of women who underwent breast augmentation with silicone gel implants. As a consequence there are voices that call for regular checks of the implants with regard to their integrity. All the external methods of judging an implant, including MRI, can--if at all--detect only marked lesions or ruptures. Direct visualization of the implant up to now has been the only safe method. Direct visualization can be carried out by reopening the previous incision or by endoscopic inspection. Endoscopic evaluation needs the dexterity of all endoscopic plastic surgery procedures, but with sufficient training and experience it is a very safe, quick, and highly compliant procedure for the patient and can be combined with endoscopic biopsies and other endoscopic operations such as capsulotomies. PMID- 7484471 TI - Viability of fat obtained by syringe suction lipectomy: effects of local anesthesia with lidocaine. AB - The results of transplantation of free autologous fat obtained by blunt syringe suction lipectomy are unpredictable. We examined if adipose tissue viability is compromised by using syringe suction lipectomy and by infiltration of the tissue with local anesthetics. As reference, we used adipose tissue samples excised during elective surgery. Fat obtained intraoperatively and by lipectomy was digested with collagenase to isolate adipocytes. The mechanical damage associated with sample handling and cell isolation in both procedures was similar and did not exceed 6% of the total cell mass. In addition, cells isolated from intraoperative and lipectomy samples did not differ functionally, responded similarly to insulin stimulation of glucose transport and epinephrine-stimulated lipolysis, and retained the same growth pattern in culture. Since during fat transplantation the graft is exposed to local anesthetics at both the donor and the recipient sites, we reexamined adipocyte function in the presence of lidocaine. Lidocaine potently inhibited glucose transport and lipolysis in adipocytes and their growth in culture. That effect, however, persisted only as long as lidocaine was present; after washing, the cells were able to fully regain their function and growth regardless of whether the exposure was as short as 30 minutes or as long as 10 days. These results indicate that adipose tissue obtained by syringe lipectomy consists of fully viable and functional adipocytes, but local anesthetics may halt their metabolism and growth. PMID- 7484475 TI - Residual silicone detection using MRI following previous breast implant removal: case reports. AB - The current controversy surrounding the safety of silicone gel breast implants has resulted in an increasing number being removed. Although previous reports have suggested that remnants of the implant capsule are reabsorbed after explantation surgery, the persistence of the capsule in fact may be associated with implant fragments and silicone gel leakage. In this study we have used magnetic resonance imaging (MRI) to identify residual silicone gel and silicone granulomas following the removal of silicone gel breast implants. Four representative clinical case reports are presented. These patients, who had residual silicone present in their bodies, presented to us with breast pain, palpable masses, or abnormal calcific mass densities apparent on a mammogram. High-resolution MRI images were found to be helpful in identifying local and remote collections of silicone gel, silicone granulomas, and residual capsules that were incompletely removed from previous explantation surgery. MRI breast images demonstrated high resolution and provided the accurate anatomical locations of residual silicone gel and silicone granulomas in all the regions of breast parenchyma, chest wall muscles, and axillae. Patients with persistent local symptoms following explantation surgery may benefit from an evaluation of the breast using MRI. PMID- 7484476 TI - Prevention and treatment of linear scar formation in the scalp: basic principles of the mechanism of scar formation. AB - Linear scar formation in the scalp after suturing an incision has been considered unavoidable. It was not known why scars formed even if the hair bulb was left intact. The authors developed a subcutaneous tissue-shaving method for radical treatment of bromidrosis and studied the process of hair regeneration by using thick-tissue specimens. They suggest that stem cells (lower) are located not only in the lower end of the telogen hair follicles but also in the sebaceous isthmus at the secretory opening of the sebaceous gland (upper stem cells). They found that linear scars can be prevented and existing linear scars can be surgically treated by using a relaxed suture on a scalp incision to avoid excessive pressure on the upper stem cells. PMID- 7484477 TI - Wetting solutions in aspirative lipoplasty: a plea for safety in liposuction. PMID- 7484478 TI - Cochrane Collaboration in health care of elderly people. PMID- 7484479 TI - Oxygen desaturation on swallowing as a potential marker of aspiration in acute stroke. AB - We have assessed the measurement of oxygen saturation (SaO2) as a means of detecting aspiration in patients with stroke. For 10 weeks all acute stroke [AS] admissions were seen within 48 hours. Basal SaO2 was measured by pulse oximetry. Patients swallowed 10ml water while sitting up and SaO2 was noted for 2 minutes. Two control groups [young, fit (YF) and inpatient age- and sex-matched, non neurological disease (IP)] underwent the same assessment. AS subjects underwent independent assessment of swallowing by a speech and language therapist (SLT). Exclusion criteria comprised impaired consciousness, other neurological disease and chest infection. Forty-nine AS subjects [20 men; aged 46-93 (mean 71) years], 55 YF [26 men; aged 18-55 (mean 32) years] and 65 IP [28 men; aged 53-96 (mean 71) years] were studied. Mean (SD) SaO2 fall in AS subjects [2.6 (2.9)%)] was significantly more than in YF [1.1 (0.8)%] or IP [1.1 (0.9)%]. The lower 95% confidence limit for variation in SaO2 did not differ between YF and IP (3.0% 'fall'); 19 (39%) AS subjects desaturated below this 95% lower confidence limit. Mean (SD) SaO2 fall was significantly more in SLT-graded 'aspirators' [4.6 (2.7)%] than 'nonaspirators' [1.4 (1.0)%]. We conclude that (1) a fall in SaO2 on swallowing fluid is common in patients with acute stroke; (2) the presence or absence of desaturation agrees statistically with SLT assessment of aspiration; (3) SaO2 measures may aid bedside assessment of swallowing. PMID- 7484480 TI - Life-style factors associated with winter serum 25-hydroxyvitamin D levels in elderly adults. AB - A cross-sectional survey of community-dwelling elderly volunteers (104 women, 87 men, mean age 70 years) was carried out in winter to determine whether cardiovascular and other potentially modifiable lifestyle factors were associated with serum levels of 25-hydroxyvitamin D during this period. Mean winter levels of serum 25-hydroxyvitamin D were higher in men than in women (14.2 vs. 12.2 ng/ml, p = 0.021), higher in those who used margarine daily compared with less often (14.4 vs. 11.9 ng/ml, p = 0.005), and higher in those who holidayed overseas in the last 6 months at a lower latitude than in Great Britain or Germany (14.9 vs. 12.4 ng/ml, p = 0.044), after controlling for other confounders. Serum 25-hydroxyvitamin D levels were not related to the number of hours spent outdoors during the previous month (r = 0.04, p > 0.05). When participants were classified by tertile of low-density lipoprotein (LDL) cholesterol, mean serum 25-hydroxyvitamin D was raised in the highest (14.3 ng/ml, p = 0.011) and middle (13.3 ng/ml, p = 0.15) tertiles compared with the lowest tertile (11.9 ng/ml), after controlling for other confounders. All other cardiovascular risk factors, including total- and HDL-cholesterol, blood pressure, BMI, smoking and leisure physical activity, were unrelated to serum vitamin D. These results suggest that body vitamin D levels in elderly people during winter are related to diet and overseas holidays within the last 6 months, but not to UK winter sun exposure, nor to the major cardiovascular risk factors apart from LDL cholesterol. PMID- 7484481 TI - Community survey of home nebulizer technique by elderly people. AB - To determine the prevalence and nature of difficulties elderly subjects experience using home nebulizers we have studied all patients in the Leeds Health District aged 70 years and older loaned a nebulizer from the hospital for at least 6 months. Of 42 subjects with a nebulizer, 40 (95%) agreed to assessment. They had a mean age of 79 years (range 70-84 years) and mean duration of nebulizer use of 38 months (range 6-120 months). Twenty subjects had one or more problems with 14/40 (35%) rated as moderate or severe; difficulties with cleaning 16/40 (40%) and filling 7/40 (18%) were the most frequent; 13/40 (33%) were sometimes or always dependent on their carers to administer their nebulizers. Elderly subjects often experience practical difficulties, or require help from a carer, when using a home nebulizer. PMID- 7484482 TI - Improvements in exercise capacity and dyspnoea by inhaled anticholinergic drug in elderly patients with chronic obstructive pulmonary disease. AB - To test the effects of the inhaled anticholinergic drug, oxitropium bromide (OTB), on exercise capacity and dyspnoea in elderly patients (more than 75 years old) with chronic obstructive pulmonary disease (COPD), we performed cycle exercise testing on 12 elderly patients with COPD [mean age 78.7 (SD 1.1) years; FEV1% 41.3 (2.0)%] as well as 12 middle-aged COPD patients [mean age 60.1 (1.0) years; FEV1% 38.9 (2.4)%] before and after inhalation of OTB or placebo in a double-blind and placebo-controlled design. Spirometry was repeated immediately before and 30 min after OTB or placebo inhalation. Dyspnoea was quantitatively evaluated using the slope of the regression line between Borg scale and oxygen uptake (VO2) during exercise (Borg scale slope: BSS). After OTB inhalation, spirometric indices, exercise capacity as indexed by maximal VO2, and dyspnoea index (BSS) were improved compared with pre-inhalation value in both the elderly and middle-aged patients. The magnitude of improvements in these indices in the elderly patients was not different from that in the middle-aged. We conclude that the inhaled anticholinergic drug produces useful improvements in dyspnoea and exercise capacity in both elderly and middle-aged COPD patients. PMID- 7484483 TI - Sustained improvement of subjective quality of life in older community-dwelling people after treatment of urinary incontinence. AB - We aimed to study the impact of conservative non-pharmacological, non-surgical management on quality of life in elderly incontinent patients. Community-dwelling patients age 60 years or older were randomized to receive immediate or delayed conservative management. A number of questions relating to quality of life were administered at admission and then at 2, 4, 8 and 12 months. By 4 months, incontinence was cured in 25% and improved in 63% of patients. The frequency and severity of incontinence was reduced (p < 0.001), and deferment time was improved (p < 0.01). There were statistically significant improvements in subjective quality of life measures at 4 and 12 months involving depression (p < 0.001), isolation (p < 0.03), embarrassment (p < 0.001), laundry (p < 0.001) and smell (p < 0.02) when comparing these variables with initial assessment. This effect did not appear to be a placebo effect as evidenced by comparison with the delayed intervention group, and was sustained over 12 months. There are significant short and long-term benefits to the quality of life of older patients with incontinence when treated by conservative measures. Intervention studies should measure and report quality of life as an outcome variable. PMID- 7484484 TI - Oxybutynin with bladder retraining for detrusor instability in elderly people: a randomized controlled trial. AB - The aim of this study was to examine the efficacy of oxybutynin plus bladder training in the treatment of detrusor instability in frail elderly patients living independently in the community. It was a randomized, double-blind, placebo controlled parallel-group trial of oxybutynin in 57 elderly patients (mean age 82.2, SD 6.06), with frequency and incontinence due to detrusor instability. After a 2-week run-in period patients received a bladder training and drug or placebo for the next 6 weeks. Outcome measures were changes in frequency and incontinence, recorded throughout on diary charts, and subjective evaluation of symptoms ('better'/'not better', and using a four-point scale 'cure' to 'no change'). Oxybutynin was superior to placebo in reducing daytime frequency [95% confidence interval (CI) of difference in change in frequencies totalled over 14 days was -27.0, -6.0; p = 0.003] and in producing subjective benefit (at day 29 only), when 24/28 (86%) patients on oxybutynin described benefit compared with 16/29 (55%) on placebo (p = 0.02). There was no difference between the groups in reduction of incontinent episodes. The median dose of oxybutynin titrated for therapeutic effect was 5 mg/day, and for placebo 10 mg/day (CI of difference 0.001, 5.001; p = 0.05). Side-effects reported were of similar frequency (50%) in the two groups. We conclude that oxybutynin with bladder training is superior to bladder training alone in reducing frequency due to detrusor instability in very elderly people living at home. PMID- 7484485 TI - Physical activity and psychological well-being among people aged 65 to 84 years. AB - This report deals with the findings of an epidemiological interview carried out among two representative samples (n = 800 + 800) consisting of persons born in 1904-13 and in 1914-23, and living at home in the year 1988 in the city of Jyvaskyla, central Finland. The participation rate was 80% (n = 1244). It appeared that overall involvement in physical exercise decreased with increasing age, especially among the women. About 50% of the subjects carried out regular walking exercise and 40% practised some form of home gynastics which was considered not to be very intensive. About 20% of the subjects were no more physically active than was essential for performing their daily activities. According to log-linear and regression models, there was a significant association between higher prevalence of depression and no regular physical exercise. Self-rated meaningfulness of life and better subjective health were also significantly related to regular and intensive physical exercise. These relationships were more obvious among the younger cohort (65-74 years). The results suggest that involvement in physical exercise may promote positive perceptions of psychological well-being among the elderly. On the other hand, psychological well-being seemed to be an important predictor for staying physically active at advanced ages. These findings are based on a cross-sectional study and therefore leave open the question of direction of causality which will be pursued in a follow-up survey. PMID- 7484486 TI - Preventing pressure sores in elderly patients: a comparison of seven mattress overlays. AB - Long-term elderly patients at risk of pressure sores as shown by superficial skin breaks in the pressure areas were randomly allocated to seven low-priced mattress overlays, alternating pressure (AP): Large Cell Ripplebed; constant low pressure (CLP): Preventix, a 'Groove' prototype, Modular Propad (contoured foam); Ardo Watersoft; Spenco, Surgicgoods Hollowcore Mattress Pad (fibrefills). Assessments of the patients' medical condition, nursing management and pressure areas were carried out twice weekly for a mean of 17.7 days. If the pressure areas deteriorated significantly the trials were stopped and the patients were transferred to other supports. Thirteen per cent of trials on Ripplebeds had to be stopped compared with 32%, 35% and 37% respectively on contoured foam mattresses (p = 0.0005), 47% on the water mattress (p < 0.001) and 51% and 54% on fibrefills (p < 0.0001). Respective healing rates were: 45%, 37%, 24% and 20% (p = 0.001). Although the contoured foam mattresses provided better protection than the fibrefills (p < 0.01), only the AP mattress effectively prevented and healed sores in these patients who could not be regularly repositioned. PMID- 7484487 TI - Zinc, biochemical markers of nutrition, and type I osteoporosis. AB - Having observed previously that the reduction of levels of biological markers of nutrition in postmenopausal osteoporosis may be related to zinc deficiency, we measured plasma and urinary zinc concentrations in 30 women with postmenopausal osteoporosis and in 30 healthy postmenopausal women who served as controls. Plasma zinc levels did not differ between groups, but urinary zinc excretion was significantly higher in the women with postmenopausal osteoporosis (p = 0.002). The relation between total body bone mineral content corrected for body weight (TBBMC/W) and markers of nutrition was significant (multiple regression analysis: p < 0.0001) in the women with postmenopausal osteoporosis but not in the healthy postmenopausal controls. Likewise, the relation between TBBMC/W and plasma and urinary zinc levels also was significant in the women with postmenopausal osteoporosis but not in the controls (multiple regression analysis: p = 0.0022). Neither group showed any correlation between plasma or urinary zinc concentrations and levels of biological markers of nutrition. Urinary zinc concentration correlated significantly with serum tartrate-resistant acid phosphatase level (simple linear regression analysis: r = 0.583, p < 0.001) in the women with postmenopausal osteoporosis but not in controls. TBBMC correlated with urinary zinc concentration significantly in the women with postmenopausal osteoporosis (simple linear regression: r = 0.567, p = 0.0015), but the correlation was nonsignificant in healthy postmenopausal controls. These findings indicate that the elevation of urinary zinc elimination in osteoporosis is dependent on bone resorption. PMID- 7484488 TI - Mobility after proximal femoral fracture: the relevance of leg extensor power, postural sway and other factors. AB - We studied 40 previously healthy women (median age 83 years 90% central range 77 90) 1 week after surgical fixation of their proximal femoral fracture to establish the significance of leg extensor power, postural sway, age, pre-injury mobility and fracture type in recovery of walking and stair-climbing ability. Subjects demonstrated a range of walking and stair-climbing abilities. Those not mobile all had sway greater than 40.9 cm/1 min and were more likely to have suffered a trochanteric fracture (p < 0.05). These subjects were not the oldest or those with the worst pre-injury mobility, but did report much greater pain (p < 0.05). Multiple linear regression revealed leg extensor power in the fractured leg (LEPf) to be the most important determinant of walking speed (R2 0.40, p < 0.001) and stair climbing time (R2 0.33, p < 0.002). LEPf was less than in the uninjured leg (p < 0.001), and accounted for only 30% of the total power output. LEPf was determined by pain, and not pre-injury mobility or fracture pattern. These findings have implications for the development of rehabilitation programmes. PMID- 7484489 TI - Haemodynamic response to head-up tilt in elderly hypertensives and diabetics. AB - To examine the haemodynamic mechanism associated with postural hypotension (PH) in elderly people, haemodynamic response to head-up tilt was studied in 14 elderly hypertensives and 13 elderly diabetics. Hypertensives and diabetics were divided into those with or without PH, defined as > or = 10 mmHg decline in mean blood pressure in response to head-up tilt. There was no significant change in cardiac output in hypertensives or diabetics with PH, whereas there was a significant increase during tilt in hypertensives without PH (p < 0.0001) and diabetics without PH (p = 0.0054). Hypertensives without PH showed a significant decrease in total peripheral resistance in response to head-up tilt (p = 0.0043). In hypertensives with PH and in both diabetic groups, there was no change in total peripheral resistance in response to head-up tilt. There was no difference in ejection fractions or heart rate responses between subjects with and without PH in either disease group. The difference in cardiac output change was not explained by myocardial changes observed at echocardiography nor by heart rate response nor by differences in total peripheral resistance. The results suggest that an increase in cardiac output in response to changing posture may be more important than vasoconstriction in protecting elderly subjects from PH. PMID- 7484490 TI - Xamoterol improves the control of chronic atrial fibrillation in elderly patients. AB - Twenty digitalized elderly patients with chronic atrial fibrillation were randomized into a double-blind cross-over study. None was in overt heart failure and all were taking < 80 mg frusemide daily. They received xamoterol 200 mg b.d. for 2 months with their usual dose of digoxin for 1 month and placebo digoxin for the other month. Twenty-four-hour heart rate analysis was done at baseline and at the end of each treatment period. Compared with baseline digoxin, xamoterol alone significantly increased nocturnal minimum heart rate [85 +/- 17 vs. 62 +/- 9 (mean +/- SD), p < 0.0001] without affecting daytime maximum heart rate (132 +/- 18 vs. 122 +/- 20, p = NS). Compared with baseline digoxin, xamoterol plus digoxin significantly increased nocturnal minimum heart rate (68 +/- 8, p < 0.05) and reduced daytime heart rate (114 +/- 17, p < 0.05). The mean number of pauses > 1.5 s was significantly reduced by xamoterol alone. Walking distance in 6 minutes was 406.1 +/- 27.1 m (mean +/- SE) at baseline and improved significantly on both treatments (450.3 +/- 19.8 on xamoterol; p < 0.02 and 453.7 +/- 19.2 on xamoterol plus digoxin; p < 0.01). No significant change was found in subjective measurements of palpitations, breathlessness and well-being using visual analogue scales. Xamoterol combined with digoxin improves effort tolerance and heart-rate control by reducing diurnal tachycardia and nocturnal bradycardia and pauses. PMID- 7484491 TI - Reduced bone density of the hip in elderly patients with Parkinson's disease. AB - Parkinson's disease (PD) is a major cause of falls and reduced mobility in elderly people. Fractures often occur as a result of these falls but there have been no studies of bone mineral density (BMD) in PD. We have prospectively studied 29 elderly women (mean age 77 years) and 26 elderly men (mean age 74 years) with PD over 2 years and compared them with similar numbers of age- and sex-matched controls. Bone mineral density was measured by dual X-ray absorptiometry. Women were more likely to sustain fractures (38% compared with 8% of men). Mean BMD of the lumbar spine was similar in the 55 combined PD patients compared with combined controls. However, the total hip BMD was 10% lower (p = 0.014) and the neck of femur BMD 12% lower (p < 0.004) in the 51 PD patients compared with controls. The relatively lower hip BMD in elderly PD men and women combined with their abnormal gait and tendency to fall backwards or to the side may explain their excess risk of hip fractures, particularly in women. The neuromuscular deficit in PD may differentially reduce hip BMD relative to other sites and may thus be amenable to measures designed to improve mechanical loading in the lower limbs in the early stages of PD. PMID- 7484492 TI - Maximal muscle strength of elderly women is not influenced by oestrogen status. AB - It has recently been reported that oestrogen replacement may preserve the ratio of muscle strength to cross-sectional area in the hand in addition to its beneficial effect on bone mass in postmenopausal women. Since muscle strength is associated with falls and fractures in elderly people, this additional effect of oestrogen replacement could be of considerable benefit to postmenopausal women. We therefore wished to determine whether this effect was also evident in larger muscle groups that are involved in ambulation, balance, and activities of daily living. We examined the relationship of oestrogen replacement therapy (ORT) to maximal dynamic muscle strength in 85 healthy women aged 65-82 years. Thirty seven women were currently taking ORT and had been on stable doses for 2 to 43 years [17.8 (1.8); mean (SEM)], while 48 women had no previous exposure to postmenopausal oestrogen therapy. Dynamic muscle strength was determined for five standard lower-body exercises using isotonic equipment by the 1-RM method. Bone mineral density (BMD, g/cm2) of the axial and appendicular skeleton as well as body composition was assessed by dual-energy X-ray absorptiometry. There was no difference between subjects according to ORT status for age, height, weight, lean body mass (LBM), fat mass, or percentage body fat. BMD of the spine (L2-4), mid radius, and whole body was greater (p < 0.001) in individuals receiving ORT, with no significant difference at the femoral neck. No differences existed between groups for lower-body muscle strength. Normalizing muscle strength to body mass and LBM did not alter the result.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7484493 TI - The association of health management with the health of elderly people. AB - To assess the effect of health management on the health of a community-based sample of people aged 65 years and over, a cross-sectional survey was carried out of 1491 people aged 65 years and over (22.3%) randomly drawn from the computerized age-sex register in Settsu City, Osaka. A total of 1473 people were contacted and complete responses were obtained from 1383 of these (a response rate of 93.9%). The proportion of the sample who had been receiving regular health checks or had been taking care of their health decreased with a decline in Activities of Daily Living (ADL). This trend was more pronounced among the younger subjects aged 65-74 years. For disability, those who had been receiving regular health checks had lower overall severity scores in both the 65-74 years and 75 years and over age groups. There were no significant differences in the severity scores for those practising health maintenance (watching diet and exercise) in both age groups, but those practising health maintenance tended to have less disability. The percentage of those who were socially active and thought life worth living also decreased with the decline in ADL for both age groups. These psycho-social factors were also associated with a lower overall severity score. A multiple linear regression analysis showed that health checks, social activity and the presence of aspects of life subjectively evaluated as making it worth living (Ikigai) were shown to have statistically significant negative associations with disability when other factors were controlled for. PMID- 7484494 TI - Review: urinary tract infection in elderly people--time to review management? PMID- 7484495 TI - Accidents involving older people: a review of the literature. PMID- 7484496 TI - Evaluation of sympathetic function in elderly subjects. PMID- 7484497 TI - Collaboration with orthopaedic surgeons. PMID- 7484498 TI - The costs and effects of early discharge in the management of fractured hip--a correction. PMID- 7484499 TI - [Fundus hypothermia at 29 degrees C prevents ischemic injury of the outer retina]. AB - We evaluated quantitatively the protective effect of local fundus hypothermia under pressure-induced ischemia using morphometric analysis. Retinochoroidal ischemia was produced in albino rabbit eyes by increasing the intraocular pressure for 60 minutes. During the ischemic procedure, a copper plate was inserted behind the eyeball. The retinal temperature in the posterior pole was thus reduced to 29 degrees C by placing solid carbon dioxide, and to 32 degrees C by placing an ice cube at the anterior end of the plate. Histopathological changes in the group with ischemia alone were obvious in visual cells and retinal pigment epithelial cells (RPE), but the retina treated with additional hypothermia was well preserved. In the retina with hypothermia at 29 degrees C, there was no significant difference from the controls in the mean thickness of the photoreceptor layer (PRL) and the RPE, and the average count of nuclei in the outer nuclear layer (ONL). In the retina with hypothermia at 32 degrees C, there was also no significant difference from the controls in the thickness of the PRL and the RPE. Otherwise, the count of nuclei in the ONL decreased significantly when compared to that of controls (p < 0.001). These findings indicate that even mild hypothermia at 29 degrees C preserves the outer retina from ischemic damage and that the protective effect of hypothermia at 32 degrees C is insufficient. PMID- 7484500 TI - [A battery model of the eyeball to calculate standing potential of the eye]. AB - Variation of the resting potential caused by eye movements was analyzed by applying a battery model to the eyeball. A formula that gives the potential between the inner canthus and the outer canthus of the eye was derived on the basis of the electric theory in electromagnetics, on the assumption that the eyeball was floating in a homogeneous conductor. The variations of the resting potentials were represented using the distance from the center of the eyeball to the potential detecting position and the deflection angle of the position with respect to the optical axis of the eyeball. To verify the validity of the formula, the variation of the resting potential caused by the various amplitudes of bilaterally symmetrical eye movements were measured. The calculated results were found to be in good agreement with the experimental ones. PMID- 7484501 TI - [The relationship between visual field defects and clinical factors in normal tension glaucoma]. AB - We investigated the relationship between visual field defects and some clinical factors in 30 eyes of 30 normal-tension glaucoma (NTG) patients. The static central visual field by the Humphrey field analyzer, diurnal variation of intraocular pressure (diurnal IOP), blood flow velocity of the ophthalmic artery by color Doppler imaging, blood flow volume of digital tissue by laser flowmetry, and systemic blood pressure (BP) were measured after rinsing out the anti glaucoma drug for one month. Mean deviation (MD) calculated by the Humphrey STATPAC program as an index of visual field defect, was correlated to the Pourcelot index (RI) of ophthalmic arterial blood flow and magnitude of diurnal IOP. In multiple regression analysis, the multiple correlation coefficient was 0.79 and the coefficient of determination was 0.62. RI, systolic BP, magnitude of diurnal IOP, and the refraction and mean of diurnal IOP were used in the multiple correlation model as factors significantly related to MD. These results suggest that both IOP and some clinical factors may contribute to the visual field defects of NTG. PMID- 7484502 TI - [The time required topical carbonic anhydrase inhibitor (MK-507) to take effect on intraocular pressure of normal human eyes]. AB - The effective time course of a topical carbonic anhydrase inhibitor, MK-507, on intraocular pressure (IOP) was studied in 6 normal volunteers. As a baseline study, IOP was determined and aqueous protein concentration was determined by laser flare photometry every half hour for 5 hours with the instillation of a placebo. Then, an examination with the same time course was performed on the same subjects with the instillation of 1% MK-507 or oral administration of acetazolamide at a dose of 250 mg. MK-507 lowered IOP 30 minutes after the instillation, and showed a significant reduction of IOP at 1 hour. Aqueous protein concentration increased significantly 30 minutes after the MK-507 instillation. Acetazolamide lowered IOP 30 minutes after the administration, and showed a significant reduction of IOP at one hour. Aqueous protein concentration increased significantly one hour after the acetazolamide administration. Corneal thickness, anterior chamber depth, anterior chamber volume, and serum protein concentration were not significantly changed by the drug treatment. These results suggest that the inhibition of aqueous humor production causes the rapid reduction of IOP seen in the use of MK-507 as well as acetazolamide. PMID- 7484503 TI - [Traumatic macular hole and posterior vitreous]. AB - We studied retrospectively the clinical features of 18 eyes of 18 patients with traumatic macular hole. The macular hole was elliptical with irregular edges in 17 eyes (94%) and its size ranged from 1/5 to 1/2 of the disc diameter. Posterior vitreous detachment was found in 3 eyes (17%); the vitreous was detached from the macula in only one of these 3 eyes. Commotio retinae, vitreous hemorrhage, hyphema, or choroidal rupture was also found in several eyes. These findings suggest that traumatic macular hole develops usually in the absence of posterior vitreous detachment and its pathogenesis is independent of the occurrence of posterior vitreous detachment. We speculate that blunt trauma-induced deformity of the eyeball or impact on the posterior pole provides a mechanical cause for the macular rupture. PMID- 7484504 TI - [Effect of tilting on cyclodeviation in nine diagnostic positions of gaze in normal subjects with Awaya's New Cyclo Tests]. AB - We measured cyclodeviation in all nine diagnostic positions of gaze in normal subjects with the New Cyclo Tests and phase difference haploscope. Measurements with the New Cyclo Tests were carried out in the upright posture, and head tilt 15 or 30 degrees to the right or left. Measurements with the phase difference haploscope were carried out in the upright posture with or without the fusional background. Cyclodeviation in all nine diagnostic positions of gaze measured with the New Cyclo Tests in the upright posture was similar to that measured with phase difference haploscope under the fusional background. Both tests showed that excyclodeviation was significantly greater in upward, right-upward and left upward gaze than in the primary position. It is concluded that cyclodeviation measured with the New Cyclo Tests is not affected by cyclophoria. The amount of excyclodeviation measured with New Cyclo Tests showed no difference between head tilt postures. We suggest that the effects of head tilting on cyclodeviation could be compensated by the corresponding effect on cyclotorsion of both eyes in normal subjects. PMID- 7484505 TI - [Surgical and botulinum toxin treatment in two cases of abnormal retinal correspondence-exotropia with congenital homonymous hemianopsia]. AB - We report 2 unusual cases of congenital occipital hemianopsia associated with abnormal retinal correspondence (ARC)-exotropia. Two Japanese males, 25 and 28 years of age, visited our hospital for surgical correction of manifest exotropia with dissociated vertical deviation and overaction of the superior oblique muscle. Visual field examination demonstrated homonymous hemianopsia with approximately 5 to 10 degrees of macular sparing. Visually evoked potential examination showed small amplitude similar to dissociated vertical deviation patients in all half-field stimulation. Electrooculogram examination demonstrated defective pursuit to the ipsilateral side. Some investigators have speculated that progressive exotropia compensates for homonymous hemianopsia and is a rare contraindication for strabismus surgery. To confirm the deterioration of motor and visual functions before surgery in these patients, we tried injections of botulinum A type toxin into the lateral rectus muscle ipsilateral to the hemianopsia. Contrary to the hypothesis, our patients had no change in their binocular visual fields and visual function. Finally, we performed recession of the lateral rectus muscle, so that both patients were satisfied with their ocular alignment, with no marked change of visual behavior. PMID- 7484506 TI - [Relationship between kinetic visual acuity and visual acuity with limited exposure]. AB - Using a kinetic vision tester which we developed, kinetic visual acuity (KVA) at target velocity of 0-100 km/h was measured at background luminance of 0.1-200 cd/m2 in 19 eyes of 10 normal volunteers ranging in age from 20 to 31 years. The decrease in KVA was slow at a target velocity of 30 km/h or more as reported before. KVA was best at background luminance of 100 cd/m2. In 3 eyes of 2 normal subjects, 3 eyes of 2 patients with tonic accommodation and 1 eye of 1 patient with central serous chorioretinopathy, visual acuity at limited exposure times was also measured. In patients with tonic accommodation, visual acuity at limited exposure times was good, but KVA was poor as compared to normal eyes. In the eyes of patients with central serous chorioretinopathy, results were poor for both. The quantity of light required to obtain the same visual acuity was similar for both normal eyes and eyes with central serous chorioretinopathy in terms of both KVA and visual acuity at limited exposure times. In eyes with tonic accommodation, difference was seen in the required quantity of light. We conclude that functions of accommodation are involved in kinetic vision and that higher visual functions are required than those for visual acuity at limited exposure times. PMID- 7484508 TI - [Normal-tension glaucoma and primary open-angle glaucoma. Are they different entities?]. PMID- 7484507 TI - [A case of uncomplicated retinal astrocytoma]. AB - A 45-year-old Japanese women whose left eye contained an elevated, whitish yellow, and multi-lobed mass of 1.5 disc diameters that extended out from the retina adjacent to the optic disc. Ultrasonography demonstrated high reflectivity corresponding to the lesion of the left eye. By fluorecein angiography the tumor showed autofluorescein, retinal vessel through the tumor in the venous phase, and in the late phase pronounced staining of the tumor. A scanning laser ophthalmoscope showed the surface in detail. A search of the literature revealed only 16 cases of uncomplicated retinal astrocytoma. PMID- 7484510 TI - [The effect of steroids on organ-cultured porcine trabecular meshwork--an ultrastructural, biochemical, and lectin histochemical study]. AB - The effect of corticosteroid administration on porcine trabecular meshwork (PTM) was studied by ultrastructural and biochemical analysis. Electron microscopic observations showed an increase of elastic fibers in PTM treated with steroids. Two-dimensional polyacrylamide gel electrophoresis with silver staining showed an increase of proteins with molecular weights of 52 kDa, 66 kDa and 70 kDa. The 66 kDa protein was assumed to be a glycoprotein containing N-acetyl-D-glucosamine, as it was stained positively in the lectin blot analysis with Triticum vulgaris (WGA) and Datura stramonium (DSA). The elastic fibers reacted positively to WGA and DSA, and the degree of this reaction was strengthened by steroid treatment. These results demonstrated that steroid treatment induces ultrastructural and biochemical changes in PTM, and an increase of elastic fibers containing glycoprotein with N-acetyl-D-glucosamine. PMID- 7484509 TI - [The effect of corneal epithelial cells on the collagen gel contraction by keratocytes]. AB - To understand the mechanism of corneal stromal wound contraction, we investigated the effect of corneal epithelial cells on the collagen gel contraction by keratocytes. Subcultured rabbit keratocytes embedded in type I collagen gel were cultured. Rabbit corneal epithelial cells were also cultured and the cultured medium was collected and used as an epithelial cell conditioned medium (ECCM). The collagen gel contraction by keratocytes was estimated by measuring the diameter of the collagen disc once a day for 5 days. The diameter of the collagen gel decreased in proportion to the number of keratocytes in the presence of fetal calf serum (FCS), but it decreased in inverse proportion to the concentration of collagen. The collagen gel contraction by keratocytes was enhanced by the addition of ECCM in a dose dependent manner. This stimulatory activity was found in ECCM obtained from 3 to 7 day cultivation of corneal epithelial cells. These findings demonstrated that keratocytes contracted collagen gel in the presence of FCS. It also appeared that corneal epithelial cells secrete a factor or factors that stimulate the collagen gel contraction by keratocytes. PMID- 7484511 TI - Osteoma of the mandible. A case report. AB - The authors present a case of osteoma of the mandible. Osteomas are solitary lesions occurring predominantly in the skull and many authors believe that most osteomas in the maxillofacial areas are reactive bone hyperplasia or advanced ossifications. In some countries the prevalence of the osteoma is about 1% of all patients in the ENT clinics. Malignant transformation has not been described and surgery of the lesion should be conservative and consist essentially of enucleation. PMID- 7484512 TI - [The role of microsurgery in the reconstruction of loss of substance of soft tissue in the maxillofacial region]. AB - Reconstruction of oncological defects in the maxillofacial region still remains a real surgical challenge due to the complexity and the tridimensional aspect of these defects responsible for heavy functional impairment. Microsurgery allowing the transfer of free flaps gives the surgeon the opportunity to adapt a specific reconstruction to each defect and to reinsert the patient in a quasi normal social life. These reconstruction technics are routinely used in many centers; they must be integrated in our therapeutic arsenal and in the training of the maxillofacial surgeon. Seven years ago, we incorporated microsurgical reconstruction in our therapeutical concepts and today, it seems to us important to share our experience of microsurgical reconstruction in the field of soft tissues defects in the maxillofacial region. PMID- 7484513 TI - Soft palate metastasis from a small cell carcinoma of the lung. AB - Metastases in the oral soft tissues are rare. They usually occur in the tongue and in the gingiva. The most common primary sites for lesions metastasizing to the oral tissues are lung, kidney, stomach, liver. The lung tumors are in the majority squamous cell carcinomas or adenocarcinoma; only rarely the primary tumors are small cell carcinomas. Small cell carcinoma is a highly malignant neoplasm with often extrathoracic metastases at the time of diagnosis, and a very low 5-years survival. Metastases from an occult lung carcinoma should always be considered in the differential diagnosis of small cell carcinomas in extrapulmonary sites. PMID- 7484515 TI - Cementum growth in impacted teeth. AB - The authors present a study on the cementum of impacted teeth. It was not possible to correlate the thickness of the cementum and the age of the patients. Significant differences were found, on the other hand, between the thickness and distribution of the cementum in different teeth. Third molars showed in almost all cases an increase in the thickness of cementum. PMID- 7484514 TI - Comparative study of results of electronic axiography with results of magnetic resonance imaging including MRI-assisted splint therapy. AB - The most common temporomandibular joint disturbance is the internal derangement. Its prevalence has been shown to be as high as 28%. The purpose of this study was on one hand to compare the diagnostic efficiency of an electronic axiographic system with magnetic resonance imaging, and, on the other hand to evaluate the results of MRI-assisted treatment monitoring of occlusal splint therapy. The results of this survey clearly show that in evaluation of temporomandibular joint disorders, electronic axiography and magnetic resonance imaging should be used in conjunction with one another to increase the accuracy of positive and differentiated diagnoses. The medical concept of disc recapture involving the use of occlusal splints seems to be merely a clinical term which is not necessarily hinged on anatomical intraarticular changes. PMID- 7484517 TI - Total androgenic blockade: a review. PMID- 7484518 TI - T-lymphocyte subsets in the peripheral blood of patients with renal cell carcinoma. AB - We analyzed the phenotypic distribution of the major T-lymphocyte subsets in the peripheral blood of 22 renal cell carcinoma patients before and after nephrectomy. We also compared these distributions to those of healthy controls and related them to tumor spread. The lymphocyte subsets were analyzed by means of flow cytometry. The CD4+ subset was the only one which was significantly decreased (x +/- SD: 36.2 +/- 6.4 vs 41.4 +/- 2.2 in healthy controls, P < 0.05), this contributing to a significant decrease in the CD4/CD8 ration (x +/- SD: 102 +/- 0.56 vs 1.42 +/- 0.28, P < 0.05). The decrease was more apparent with advanced stage. Helper cell activity increased significantly after nephrectomy in the higher stage patients (x +/- SD: 41.1 +/- 4.6 vs 36.4 +/-3.1 and 38.2 +/- 3.9 vs 30.1 +/- 4.2 for stage T3 and T4 patients respectively). Knowledge of peripheral blood lymphocyte patterns that correlate with various tumor stages and therapies could provide information in host-tumor relationship and predict the clinical outcome and response to therapy. PMID- 7484516 TI - Prognostic factors in advanced prostatic carcinoma treated with total androgen blockade. Flutamide with orchiectomy or with LHRH analogues. A Belgian multicentric study of 546 patients. AB - Prognostic factors were evaluated in advanced loco-regional (M0) or distant metastatic (M1) prostatic carcinoma treated with total androgen blockade (flutamide with either orchiectomy of LHRH-analogues), in 546 patients from a Belgian multicentric study. After a mean follow-up of 16.5 months (maximum 37 months) 113 (21%) patients had progressed (90 were patients with M1 disease (31%)). The estimated median progression-free survival exceeded 37.5 months. The results of a univariate analysis show that the following parameters are important prognostic factors with respect to progression-free survival in these patients: M stage, G grade, ECOG performance status, weight loss, concomitant disease, pain, dysuria and haemoglobin (Lee-Desu test, p < or = 0.01). From a multivariate analysis (Cox regression) the following prognostic factors were indicative of a decrease in progression-free survival: M1 stage, high initial G grade, ECOG performance status > I, high serum PSA, presence of concomitant disease, presence of pain and absence of dysuria. Age did not appear to be a statistically significant prognostic factor. PMID- 7484519 TI - Dysfunctional voiding: a complex of bladder/sphincter dysfunction, urinary tract infections and vesicoureteral reflux. PMID- 7484520 TI - A prostaglandin synthesis inhibitor, diclofenac sodium in the treatment of primary nocturnal enuresis. AB - Various treatment modalities have been used in Primary Nocturnal Enuresis (PNE) and pharmacotherapy is widely accepted. Prostaglandins increase detrusor pressure, decrease urethral pressure and lead to sodium excretion. They also antagonize hydro-osmotic effect of vasopressin by competing with this hormone. According to these functions of prostaglandins it is suggested that inhibition of prostaglandin synthesis may have value in the management of PNE. We evaluated the efficacy or oral diclofenac sodium treatment in 78 patients. We conclude that diclofenac sodium, an inhibitor of prostaglandin synthesis, is a good alternative agent for nocturnal enuresis particularly as a supplementary treatment combined to Imipramine, with 60% complete response and 13.3% recurrence rate. PMID- 7484521 TI - Urodynamic effects of Prolase 2 laser prostatectomy in benign prostatic hyperplasia. AB - This prospective study was undertaken to evaluate the safety and efficacy of visual laser ablation of the prostate with the Prolase 2 sidefiring neodymium: YAG laser fiber in patients with symptomatic bladder outlet obstruction due to benign prostatic hyperplasia (BPH). The Prolase 2 fiber is a non-contact sidefiring fiber, producing coagulation of the prostatic tissue. Our study describes the results obtained in 30 patients who underwent VLAP with a mean follow-up of 6 months. The evaluation of symptoms according to the WHO International Prostate Symptom Score and Quality of Life Score (IPSS and QL) demonstrated a significant subjective response rate (p < 0.001). Objective assessment of treatment with peak and average urinary flow rates, post-void residual urine volumes and pressure-flow urodynamic studies showed a significant improvement after treatment (p < 0.001). Ultrasonic assessment of prostatic volumes at 6 months showed a mean decrease in total volume of 37% after laser prostatectomy. The early and late complications of the treatment are presented but overall toxicity is low. Laser prostatectomy performed with the Prolase 2 fiber seems to be safe and efficacious for BPH patients but long term efficacy and retreatment rates have to be awaited. PMID- 7484522 TI - Visual laser ablation of the prostate (VLAP): an effective alternative treatment of benign prostatic hyperplasia. AB - A total of 75 patients suffering from benign prostatic hyperplasia (BHP) were treated with visual laser ablation of the prostate (VLAP) using the Nd-Yag laser. Preoperative evaluation consisted in determination of urinary flow and residual urine, assessment of voiding symptoms and urodynamic evaluation. Postoperatively, the same evaluation was performed at 3 and at 6 months. VLAP is a relatively simple technique with a low perioperative morbidity and short time results are comparable to classical transurethral resection of the prostate. Therefore VLAP seems to be an effective alternative treatment method for BPH which may replace classical transureteral resection of the prostate in a selected number of cases. PMID- 7484523 TI - The contribution of the obturator nerve block in the transurethral resection of bladder tumors. AB - This study was designed to evaluate the efficacy of local block of the obturator nerve to avoid it's stimulation during transurethral resection (TUR) or tumors localised on the lateral bladder wall. Forty-five patients were studied, in whom the TUR operations were performed under spinal anaesthesia. The patients were divided into two groups: in the first group a local block of the obturator nerve was done, while the second group served as controls, and the stimulation of the nerve was avoided by reduction of the electrocoagulation voltage, incomplete bladder filling and resections of smaller ships. In the first group no stimulation was observed while in 5 patients of the second group was not possible to totally resect the tumor due to the persistent stimulation of the obturator nerve. In conclusion the local block of the obturator nerve is an effective method to avoid it's stimulation, easily performed and without complications. PMID- 7484524 TI - Quality of life assessment in heterotopic and orthotopic neobladder reconstruction: a comparison. AB - Several techniques for continent urinary diversion are developed and applied in recent years. Heterotopic or orthotopic bladder replacement is an effective treatment option in selected patients. We reviewed 45 neobladder reconstructions in 25 men and 20 women. Mean age at operation was 43.7 years, mean follow-up was 34.7 months. Indications for bladder reconstruction were transitional cell carcinoma of the bladder (18), interstitial cystitis (6), neurogenic bladder (4), refractory incontinence (3) and undiversion (14). In 27 patients an orthotopic replacement (Mainz pouch in 18 and ileal neobladder in 9) was performed, in 18 patients a heterotopic replacement (Indiana pouch) was performed. Standard radiologic evaluation included ultrasonography, intravenous urography and postoperative pouchography. Preoperative urodynamic investigation and postoperative neobladder function documentation was performed routinely. Quality of life assessment was achieved by interview and use of the sickness impact profile (SIP) questionnaire. Nineteen patients (42.2%) had a completely uneventful postoperative course. Postoperative complications were acceptable as only 2 out of 20 late complications needed an open procedure for ureteric stenosis. Peri- or postoperative mortality did not occur. The mean postoperative bladder capacity was about 600 ml; 500 ml for the ideal neobladder, 560 ml for the Indiana pouch and 700 ml for the Mainz pouch. This tendency was not statistically significant. In 34 (75.5%) patients full urinary continence was achieved. One patient was consistently stress incontinent. The final overall outcome of these reconstructive procedures was excellent in the majority of cases. PMID- 7484525 TI - [Rouxel-Coadou method of ileal neobladder: clinico-functional considerations]. AB - Twelve patients are submitted to orthotopic ileal neobladder, performed using staplers. The simplicity and speed of automatic stapling devices in the Rouxel Coadou's surgical technique are reported. All patients underwent clinical and urodynamic evaluation at 3, 6, 12 months, after surgery (mean follow-up 6 months). They preferred complete daytime continence and 2 pts/4 night-time continence too. At one year, postvoid residual volume was absent, mean neobladder capacity was 450 ml and mean pressure at maximum capacity was 30-40 cm H2O. These preliminary results indicate that this technique guarantees a low-pressure reservoir with good capacity and compliance. PMID- 7484528 TI - [Clinical investigation of grade-up superficial bladder cancer]. AB - Between January 1977 and December 1993, 249 patients with grade 1 or grade 2 superficial bladder cancer were initially treated at Yokohama City University Hospital. Eighty-six patients (33%) had recurrent tumors after initial resection, and sixteen recurrent cases were so-called grade-up tumors that is, grade 3 originating from grade 1 or grade 2 tumor. The morphology of the grade-up tumors mostly showed non-papillary and invasive type. Positivity for urinary cytology of grade-up tumors was 88%. The five-year survival rate of the patients with grade up tumors was 85% after initial resection of grade-up patients and 48% after treatment of grade-up tumors. The five-year survival rate of the patients with grade-up tumors who were treated by total cystectomy was 72%, whereas that in the patients who were treated by bladder preservation therapy showed a 13% five-year survival rate and all of the six patients died of cancer during the six-year follow-up period. These findings suggest that patients who have grade up tumors should be treated by radical treatment with radical cystectomy. PMID- 7484526 TI - A clinical diagnosis of urinary bladder (extra-adrenal) phaeochromocytoma. AB - Urinary bladder (extra-adrenal) pheochromocytoma is a catecholamine-producing tumor that arises from chromaffin cells in the extra-adrenal paraganglion system of the urinary bladder. The symptoms and signs result from the release of epinephrine and/or non-epinephrine, which is more prominent during micturition. The most serious consequences of the disease are paroxysmal hypertension during micturition and malignant degeneration. Thus, proper treatment is mandatory. PMID- 7484527 TI - [E-cadherin expression and histopathological features in renal cell carcinomas]. AB - E-cadherin, a member of the cadherin family, plays a major role in cell-cell adhesion of normal epithelium. Recent studies have demonstrated that heterogeneous expression, reduction or loss of E-cadherin is involved in invasion and metastasis of cancer cells. In this study, the localization of E-cadherin in the normal human kidney and the relationship between E-cadherin expression and histopathological features in renal cell carcinomas was examined immunohistochemically. Renal cell carcinoma tissues and normal kidney counterparts were obtained from 20 patients. E-cadherin in the normal kidney was detected in the cell-cell border of the distal tubules, collecting duct and Bowman's capsule but not in the proximal tubules. E-cadherin expression was reduced in all the clear cell type renal cell carcinomas with compact or cystic configuration (n = 15), while it was well preserved in all the papillary type (n = 3) and chromophobe cell type (n = 1) renal cell carcinomas. Different expression patterns between primary site and metastasis, i.e., homogeneously weak in primary tumor and heterogeneously positive in metastatis, was observed in a case of clear cell type renal cell carcinoma. Different patterns of expression between clear and non-clear cell type, or between papillary and non-papillary type, together with strong expression in chromophobe type might reflect the origin of each type of renal cell carcinoma. Further studies will clarify whether the change in expression of E-cadherin is associated with the prognosis of renal cell carcinoma. PMID- 7484530 TI - A case of massive adrenal malignant pheochromocytoma: management of a large pheochromocytoma. AB - A 69-year-old man was referred to our hospital for a left abdominal tumor measuring about 30 cm in diameter. Laboratory examination revealed an elevation of norepinephrine in plasma and of its metabolites in urine. CT scan disclosed a huge cystic tumor in the retroperitoneal space and an enlarged aortocaval lymph node, suggesting a diagnosis of malignant pheochromocytoma. The hemodynamic studies showed low blood volume and high vascular resistance, and therefore, he was treated with vasodilators and volume expansion. His hemodynamic status normalized and a complete excision was performed. Pathological examination revealed that the patient had a pheochromocytoma with metastasis to a lymph node. The total weight of the tumor was 5,930 g. Since pheochromocytomas can become a large with a risk of malignancy, they should be surgically excised as completely as possible with further treatment after making a definite diagnosis. PMID- 7484529 TI - [Effect of BBN on the small intestine transplanted to the urinary bladder of rat]. AB - The effect of N-butyl-N-(4-hydroxybutyl) nitrosamine (BBN) on the small intestine in rat, in which augmentation cystoplasty was performed, was examined. Seventeen rats were divided into 3 groups: 1) jejunum-transplanted group (5 cases), 2) ileum-transplanted group (6 cases) and 3) control group without operation (6 cases). A part of the small intestine was transplanted to the urinary bladder, then 0.05% BBN was given for 12 weeks. The rats were killed 30 weeks after the beginning of the administration for histopathological evaluation. Bladder tumors in various grades were confirmed in all the cases. In two cases in the jejunum transplanted group, transitional cell carcinoma was found in the urinary bladder, one of which had an invasion to the adjacent part of the jejunum to the bladder mucosa. These findings suggest that the metabolic products of carcinogens, excreted into urine, had no effect on the transplanted small intestine. PMID- 7484531 TI - [Bilateral synchronous renal cell carcinoma: a case report]. AB - A case of bilateral synchronous renal cell carcinoma is reported. The patient was a 55-year-old man. A right renal tumor was pointed out by ultrasound during a health check in July, 1994. Enhanced CT scan and angiography revealed bilateral renal tumors. Right nephrectomy and left partial nephrectomy were performed on November 25, 1994. Both tumors were completely encapsulated. Histological examinations of these tumors revealed renal cell carcinoma, Expansive type, alveolar type, clear cell subtype, G2, INF alpha, pT1, pV0. PMID- 7484532 TI - [Bilateral renal cell carcinoma of von Hippel-Lindau disease associated with von Willebrand disease: a case report]. AB - A 42-year-old man diagnosed with von Willebrand disease 7 years earlier visited his home doctor with a chief complaint of gross hematuria. Abdominal ultrasonography showed bilateral renal tumors. He was referred to our hospital on June 15, 1993. His mother also had von Hippel-Lindau disease and was treated by hemodialysis after bilateral nephrectomy. The examination and imaging revealed bilateral renal cell carcinoma of von Hippel-Lindau disease associated with von Willebrand disease. Partial nephrectomy and enucleation for a small tumor on the right side and enucleations for five tumors on the left side were performed. Histological diagnosis of all tumors were renal cell carcinoma, clear cell subtype, grade 1 approximately 2. The patient has been well without local recurrence or distant metastasis for 16 months since the operation. This is the first case of bilateral renal cell carcinoma of von Hippel-Lindau disease associated with von Willebrand disease in Japan. PMID- 7484533 TI - [A case of huge prostate cancer]. AB - An 89-year-old man with bilateral leg edema and a huge abdominal mass was admitted for further evaluation. CT scan showed a hugh prostatic mass which occupied the whole pelvis cavity accompanying multiple pelvic bone metastases. Suprapubic needle biopsy revealed that the mass was well differentiated adenocarcinoma of prostate origin. The treatment was initiated by 500 mg per day of estramustine phosphate combined with injectable LH-RH analogue 2 months later. The serum levels of tumor markers were markedly elevated at the first visit; PSA 210ng/ml, PAP 110ng/ml, gamma-Sm 800ng/ml. They became normalized 3 months after the initiation of the treatment, and the mass was reduced to 11.5% of the initial size, which lead to removal of indwelling urethral catheter. The patient and his family, however, refused further treatment and the patient died of disseminated disease 8 months later. PMID- 7484535 TI - [Symposium (2). Diagnosis and treatment of renal parenchymal mass. Chairmen's comments]. AB - Although renal cell carcinoma occupies clinically the majority of renal parenchymal mass lesions, indeterminate renal masses among renal cystic lesions and benign renal tumors have been frequently found with recent advances in renal imaging modalities. Renal mass lesions to be differentiated from renal cell carcinoma were focused on in the symposium and the points discussed were divided into four categories. 1. Indeterminate renal cystic masses, especially focused on multilocular cystic lesions and renal malignant lesions associated with acquired cystic disease of the kidney (ACDK) and von Hippel Lindau disease. 2. Angiomyolipoma and oncocytoma, as representative ones among a variety of benign renal tumorous lesions. 3. How to treat small renal masses serendipitously detected by abdominal CT and/or ultrasound, radical nephrectomy or renal parenchyma-preserving operation? 4. Feasibility of percutaneous ultrasound-guided fine needle biopsy for suspicious renal mass lesions. In this symposium, the diagnosis and treatment of various renal mass lesions other than renal cell carcinoma were mainly discussed and the important points to diagnose and treat patients with renal cell carcinoma have been clarified. PMID- 7484534 TI - [Sex cord/stromal tumor of the testis: a case report]. AB - We herein report a case of sex cord/stromal tumor of the testis. A 28-year-old man was admitted to our hospital with the chief complaint of painless swelling of the left scrotal contents. On examination, gynecomastia or swelling of superficial lymph nodes was not observed. With a diagnosis of left testicular tumor, left high orchiectomy was performed. Gross examination of the specimen demonstrated a mass measuring 20 x 18 x 14 mm within the testis. On the cut surface, the tumor was gray and was associated with focal hemorrhage and necrosis. The spermatic cord and epididymis were not involved by the tumor. Microscopically, the tumor demonstrated a mixed pattern consisting of, we thought, areas of Leydig-like cells as well as areas of Sertoli-like cells showing mild atypia. Without further treatment, the patient has remained free from the disease for over fifteen months since the operation. Fifty eight cases of sex cord/stromal tumor of the testis have been reported in the Japanese literature, but sex cord/stromal tumors, other than pure Leydig cell tumor or Sertoli cell tumor, are very rare. Only 4 cases have been reported including our case. PMID- 7484536 TI - [Diagnosis and treatment of renal cell carcinoma accompanying the cystic lesion]. AB - Approximately 15% of cases of renal cell carcinoma (RCC) present cystic configuration on radiologic and pathologic examination. The mechanisms of cyst formation in RCC are intrinsic multilocular cystic growth (multiloculated renal cell carcinoma: MCRCC), unilocular cystic growth (cystadenocarcinoma), cystic necrosis and tumor growth from the epithelial lining of a preexisting cyst. These cystic lesions accompanying RCC are often difficult to differentiate from the multiloculated renal cyst (MLC) or other benign cystic lesions such as hemorrhagic cyst and so on. Differential diagnosis of the complicated renal cystic lesions is discussed in this review. PMID- 7484537 TI - [Results of mutation analyses of von Hippel-Lindau disease gene in Japanese patients: comparison with results in United States and United Kingdom]. AB - Recently the gene responsible for the von Hippel-Lindau (VHL) disease was identified as a tumor suppressor gene. Our ongoing studies on the mutation of the VHL gene in Japanese 28 VHL families with single strand conformational analyses of DNA and Southern blot analyses revealed 6 cases of insertion or deletion, 1 cases of splice site mutation and 9 cases of missense mutation, and 3 possible intragenic deletions. Our analytical findings are essentially similar to those observed in the western countries. The VHL families associated with pheochromocytoma had the same mutational hot spot as those in the western countries. Molecular analyses of the VHL gene in the Japanese VHL disease substantially improved the understanding of this disease and its inheritance character. PMID- 7484538 TI - [Renal cell carcinoma and acquired cystic disease of the kidneys in patients on long-term hemodialysis]. AB - Since the first report in 1977 by Dunnill et al., a substantial number of articles have been written concerning the complications of acquired cystic disease of the kidneys (ACDK) and renal cell carcinoma developing in the native kidneys of patients undergoing long-term hemodialysis. The incidence has been reported to be 35% to 95% of ACDK in patients undergoing long-term hemodialysis. In addition, renal cell carcinoma has been described in 4% to 7% of the ACDK. The etiology of ACDK and renal cell carcinoma in the native kidneys of patients undergoing long-term hemodialysis remains unresolved. Because ACDK and renal cell carcinoma are usually asymptomatic, several reports described the use of imaging studies in the native kidneys of patients undergoing long-term hemodialysis and suggest a variety of management schemes to screen or monitor the status of the native kidneys. Ultrasonography is the most common screening study, but CT scan is more sensitive. Both studies are necessary to diagnose ACDK and renal cell carcinoma in the native kidneys of patients. We review our 10 cases and the Japanese literature with particular attention to the diagnosis and treatment of patients with ACDK and renal cell carcinoma. Our findings were similar to those reported by others. Patients receiving dialysis treatment for more than three years should have a baseline radiologic examination of the kidneys so that subsequent problems can be more easily identified and evaluated. The best surgical treatment for patients with ACDK and renal cell carcinoma is simple nephrectomy by flank incision in order to prevent surgical side effects, for example postoperative bleeding. PMID- 7484539 TI - [Diagnosis and therapy of incidental renal cell carcinoma]. AB - Renal cell carcinoma (RCC) especially incidentally found RCC has been markedly increased in the world due to development of ultrasound or computerized tomography (CT) instruments. Controversy existed on the nephron sparing treatment for incidental RCC. We have operated the incidental RCC using a technique of tumor enucleation shelling out with 5 mm normal surrounding parenchyma. It is important to demonstrate its pseudocapsule for indication of the procedure. For the purpose MRI (T2) is the most reliable method in comparison with angiography, CT and ultrasound. In 42 cases of incidental RCC, 22 cases were nephrectomized and 19 cases were enucleated. One was not operated on. In our cases there was no local recurrence, metastasis or impaired renal function for over one year. However, some problems still remain such as local recurrence (0% approximately 10%), multicentricity (7%) and residual renal function in the literature. Therefore, we propose the following 4 points for indications of tumor enucleation, (1) asymptomatic completely and incidentally, (2) < or = 4 cm in diameter, (3) stage 1 (T1, T2) with pseudocapsule, (4) peripheral location. PMID- 7484540 TI - [Multicentricity and concomitant tumors in renal cell carcinoma: analysis by serial section of resected kidneys]. AB - Nephron-sparing surgery was initially limited to the patients with localized renal cancer (RCC) present bilaterally or in a solitary kidney. Recently there is controversy in the indication for partial nephrectomy or enucleation to incidentally defined small RCC with normal opposite kidney. We examined the incidence of multicentricity in 43 kidneys removed for RCC with a diameter of less than 80 mm. The mean diameter of the predominant tumors was 45 mm (range 12 to 80 mm). The kidneys were serially sectioned at 5 mm intervals. Three of 43 kidneys (7%) had multiple RCC. The size of the concomitant tumors ranged from 2 to 15 mm. The multicentricity had no relation to pathological grade, stage, vascular invasion or infiltration pattern. In addition, the other 4 kidneys had 2 adenomas, 1 angiomyolipoma and 1 fibroma. Therefore we observed a 16% incidence of small renal nodules and a 7% multicentricity of RCC in the nephrectomized kidneys with normal opposite kidney. PMID- 7484541 TI - [Radiological diagnosis of renal oncocytoma]. AB - Eight patients (nine tumors) with histologically proven renal oncocytoma are presented. In all cases, differential diagnosis between renal oncocytoma and renal cell carcinoma could not be done on ultrasonography. On selective renal angiography, extended arteries surrounding the tumor margin was demonstrated in six of seven tumors without a spoke-wheel arterial supply. A sharp and smooth margin with capsule (lucent rim) could be found in five cases, and a spoke-wheel configuration of vessels could be seen in only two cases. A spoke-wheel pattern might be found with tumor growth. All tumors on computed tomography (CT scan) have a distinct margin, a smooth contour and a homogeneous appearance on contrast enhanced CT scan. The capsule and the presence of a central scar were clearly seen on T1 and T2 weighted images of magnetic resonance imaging (MRI). It is helpful to differentiate the oncocytoma from renal cell carcinoma. The modality of MRI may be useful in the preoperative diagnosis of oncocytoma. PMID- 7484542 TI - [Renal angiomyolipoma: diagnosis and treatment]. AB - In 10 years the diagnosis of renal angiomyolipoma (RAML) was made in 14 patients (male-to female ratio 1:3.7) at our institution; 1 case was associated with tuberous sclerosis (TS) and 1 case had regional lymph node involvement. A statistical study was done on data taken from 739 cases of RAML in the Japanese literature, including our cases. The male to female ratio was 1 to 3. Twenty eight percent of the cases were associated with TS. The ratio of bilateral cases to the unilateral one was 1 to 3. The main clinical signs were flank pain, abdominal mass, hematuria and fever elevation. Recently the ratio of nephrectomy has decreased to 30%. The percentage of detecting the fat component by ultrasonography (US), computed tomography (CT) and magnetic resonance imaging were 88.1%, 86.5% and 80.8% respectively. The percentages of visualizing hypervascularity, aneurysms, absence of arterio-venous shunt and onion peel appearance by selective renal angiography were 77.3%, 71.4%, 48.1% and 4.9% respectively. Small (less than 3 cm), asymptomatic, simple lesions with adipose component may be observed annually by CT and US until more experiences is gained with surveillance of these patients. Embolization was useful for emergency cases or pre-treatment of nephron sparing surgery, but insufficient by itself. As there still remain problems in the diagnosis of RAML, especially in the case of very small tumors, in the case with almost no adipose component and in the case associated with renal cell carcinoma, the diagnosis of RAML should be made synthetically including angiography. PMID- 7484543 TI - [Clinical implication of selective renal tumor biopsy]. AB - Since 1982, we have performed selective needle core biopsy under ultrasonic real time guidance for the specially indicated cases with a suspected renal tumor. Between June 1982 and April 1994, 100 renal tumor cases were examined by biopsy with a success rate of 97%. We evaluated 63 cases of renal cell carcinoma, 7 cases of transitional cell carcinoma, 2 cases of metastatic carcinoma, and 25 cases of benign lesions such as angiomyolipoma and hemorrhagic cyst. In 3 cases no tissue was obtained. No complications occurred except for dissemination in only one case of angiomyoliposarcoma. Recently incidental renal tumors tend to increase more and more, and small incidental renal tumors may have a high possibility of benign tumor compared with symptomatic renal tumor. Selective renal tumor biopsy may be valuable in the diagnosis of an incidental renal tumor which is difficult to differentiate between malignant tumor and benign tumor by several imaging methods. From our experiments and other reports, the clinical implication of renal tumor biopsy is discussed. PMID- 7484545 TI - A retrospective study of conservative treatment of gastric dilatation-volvulus in the dog. AB - A retrospective study of 103 dogs with gastric dilatation-volvulus (GDV) treated conservatively during the period 1985-1989 was performed. The date and number of recurrences, and the date and cause of death along with breed, age and sex were collected from clinic records and from a questionnaire sent to the owners (77% response). The treatment consisted of emptying the stomach with a stomach tube, gastrocentesis if necessary and treating shock. All of the affected dogs were from moderate to large sized breeds and the study showed that particular breeds appear to be predisposed to GDV. The average age was 7.2 years and there was no gender predisposition. Sixty-six per cent of the dogs survived the initial treatment and 50% of the dogs were still alive 1 month after treatment. Thirty nine dogs (71%) received recurrence between 5-760 days after initial treatment. Fifty-six per cent had recurrence within 3 months and all except 2 dogs, within 1 year. Data for the time of death, regardless of cause, was available for 68 dogs. Of these 59 (81%) died within a year after initial treatment. Conservative treatment was found to be an adequate life-saving procedure in the acute stage of GDV, but could not alone prevent recurrence of the disease. PMID- 7484544 TI - Onset of spermatogenesis in Corriedale ram lambs under extensive rearing conditions in Uruguay. AB - The present study was undertaken to estimate the time for the attainment of spermatogenesis in springborn Corriedale ram lambs under conditions of extensive grazing systems in Uruguay. Clinical (live weight, scrotal circumference, penile/preputial separation), morphological (light and electron microscopy) and endocrinological (testosterone levels) parameters were examined. Two experiments in 2 consecutive years were carried out. In Exp. I, 40 ram lambs were clinically examined, weighed, and blood-sampled at 2 week-intervals between 78 and 216 days of age. Sixteen were castrated in 3 selected periods: 132-162 (n:2), 145-175 (n:6) and 186-216 days (n:8). In Exp. II, 40 ram lambs appertaining to the next generation of the same flock were examined as above at 180-210 days of age, and castrated for morphological studies. The time for attainment of complete spermatogenesis correlated significantly with most corporal parameters (i.e. scrotal circumference (r = 0.52); testicular weight (r = 0.61), epididymal weight (r = 0.60), penile/preputial separation (r = 0.75). The age of castrated ram lambs correlated with the degree of spermatogenesis (r = 0.83), although no significant correlations were found with live weight or with levels of circulating testosterone. The histology showed major variations in the degree of development of the seminiferous epithelium. Cells undergoing degeneration were a common finding through the initial stages of spermatogenesis, coincident to the presence of sperm abnormalities and foreign cells in semen. By day 180 and onwards, both histology and seminal picture normalized. It is concluded that, under these rearing conditions, the onset of puberty (expressed as morphologically established spermatogenesis) in Corriedale ram lambs is attained at 180-216 days of age when they reach 23 cm of scrotal circumference and 191 g of testis weight. The finding of a high correlation between these parameters (r: 0.93) confirms the usefulness of the measurement of scrotal circumference during clinical examination of ram lambs in this breed. PMID- 7484547 TI - A field study on group housing of lactating sows with special reference to sow health at weaning. AB - Four farms that group-housed sows from 2 weeks of lactation until weaning (G farms) and 3 farms that kept the sows individually penned throughout the 5 to 6 week-long lactation period (C-farms), were compared in terms of sow health. All sows were crossbred Swedish Yorkshire x Swedish Landrace. The daily food ration was similar on all farms except during the group-housing period, when G-farm sows were fed ad libitum. Sows were grouped in the breeding section and kept grouped on deep litter in the dry sow section on all farms. Individual health examinations were performed at the time of weaning (+/- 4 days) on 179 G-farm sows and on 167 C-farm sows. Teat- and udder skin wounds occurred less frequently (p < 0.001) in G-farm sows than in C-farm sows. In addition, preweaning atrophy of all mammary glands occurred in 6.6% of the G-farm-sows but not in a single C farm sow (p < 0.001). This indicates that sow-piglet interactions decrease when sows are group housed. However, these differences did not occur in primiparous sows, suggesting that the relation between the primiparous sow and her litter is not affected. Mastitis frequency was the same in the 2 systems. Moreover, the frequency of locomotor disorders was the same in the 2 groups, and hoof overgrowth was common in both systems. These similarities could be due to the fact that all farms group housed dry sows on deep litter. A strong relation (p < 0.001) between hoof overgrowth and locomotor disorders was evident. Low access to food due to low rank among primiparous group-housed sows was indicated by a lower (p < 0.05) backfat thickness compared with multiparous sows, and a higher (p < 0.001) frequency of skin wounds compared with individually housed primiparous sows. PMID- 7484548 TI - Methods and errors in measurements of synovial fluid volume in stifles with low volume and high viscosity synovial fluid. An experimental study in goats. AB - Synovial fluid (SF) volume was calculated using various methods in the stifles of goats, in which the cranial cruciate ligament had been transected on one side. Measurements were performed prior to surgery and again 4, 8, and 18 weeks following surgery, by measuring the dilution of an injected radioactive tracer diluted by the SF. Later, 7 months following surgery, SF volume measurements using simple arthrocentesis were performed on stifles in 9 of the goats, and the SF that could not be aspirated, was calculated using 2 indirect methods simultaneously on identical fluids in 3 of these goats. SF was also collected directly during staged arthrotomy of the stifles in 4 goats. There were conflicting results between methods, but the resulting calculated SF volumes seemed to be larger in the operated stifles compared to the controls for all the methods at about the same degree. The 2 indirect methods used to calculate the fluid remaining in the joints following arthrocentesis gave disparate volume calculations. The experiments revealed sources of error in all methods. Direct methods failed to acquire the total fluid volume, and indirect methods were subject to improper mixing and escape of the injected fluid or synovial fluid or both. It was concluded that none of the methods could be used to measure the "true" volume of SF, if such a concept exists and can be defined. None of the methods were considered reliable to compare volumes in different type of joints containing this type of fluid. It was, however, concluded that all the methods gave indication of increased SF volume present on a relative basis when paired joints were compared. PMID- 7484546 TI - Housing of pregnant sows in loose and confined systems--a field study 1. Vulva and body lesions, culling reasons and production results. AB - A field study was carried out in 18 herds with loose housing of pregnant sows (loose herds) and in 18 herds with tethered or stalled pregnant sows (confined herds). Three of the loose herds were excluded due to different kind of flooring from the rest of the herds. The remaining 15 herds had partly slatted concrete floors and electronic sow feeding. The frequencies of sows with vulva and body lesions, thin sows as well as the culling reasons and production results were used as animal welfare indicators for the herds. Vulva lesions were found only in the loose herds and the mean prevalence proportion of sows with lesions within these herds was 15.2%. All vulva lesions observed in this study seemed to be caused by biting. The relative risk of vulva lesions was 2.6 times higher in the loose herds with no roughage feeding as compared to loose herds with appetite feeding of roughage. The sows in the loose herds, that had a feeding station with a mechanical hind gate had 1.8 greater risk of vulva lesions than sows in the loose herds that used a feeding station with an electronic gate. The mean prevalence proportion of sows with body lesions was 13.1% in the loose herds and 4.0% in the confined herds. Aggression between sows seemed to be the main cause of body lesions in the loose herds, while decubitus ulcers on the shoulders were the main cause of body lesions in the confined herds. Sows in loose herds that were not fed additional roughage feeding had 1.7 times greater risk of body lesions than sows in herds that used additional roughage feeding. The main culling reasons and production results were similar in the loose and confined herds. This study showed that there were welfare problems both in confined and loose herds, however, with improved management, many of the welfare problems associated with loose housing can be reduced. PMID- 7484549 TI - Equine sarcoids. A clinical and epidemiological study in relation to equine leucocyte antigens (ELA). AB - Associations between clinical parameters of sarcoids and the equine leucocyte antigen system (ELA) were analysed for 120 Swedish horses. Median age of affected horses was 5.2 years, and the majority presented with solitary tumors between 2 and 5 cm in diameter and ventral abdomen was a predilection site. Clinical signs first appeared at a median age of 3.5 years, and sarcoids at different locations first appeared at different ages. Lesions at different sites differed in size, and multiple tumors, early onset, long duration, and older age all had an association with large size. Clinical manifestations of sarcoids and the association between certain ELA-specificities and early onset (A5) and increased recurrence rates after surgery (W13), in addition to increased prevalence (A3W13), strengthen further that some horses are inherently predisposed to sarcoid growth. Unassociated with any clinical parameters, one third of the untreated horses became free of sarcoids due to "spontaneous" regression, perhaps as a result of immune responses against the tumors. Seventy percent of the horses were treated (mostly by excision), and large size was the main parameter promoting treatment. Excision had no significant effect on possibly remaining sarcoids. Recurrence rate after first treatment was about 35%, with the majority of tumors recurring within 4 months. Early onset, long duration, large size, and localization to distal limbs all appeared to increase risk of recurrence. Early treatment, performed under general anesthesia in recumbency which permits wide excision and measures to avoid autoinoculation, significantly reduced recurrence rates. PMID- 7484550 TI - A comparative study of Staphylococcus aureus strains isolated from bovine subclinical mastitis during 1952-1956 and 1992. AB - Fifty-two strains of S. aureus isolated from cases of bovine subclinical mastitis in 52 different dairy herds in Denmark, in the periods 1952 to 1956 and 1992, were compared with regard to their phage- and EcoRI ribotypes. Furthermore, susceptibility to penicillin and production of fibrinolysin were used as additional phenotypic markers. Forty-nine strains (94%) could be separated into 12 phage types. Ribotyping assigned the 52 strains to 21 different types. Both methods showed that 57% of the 1950's strains and between 38-45% of the 1992 strains belonged to 3 dominating types. The remaining strains were placed by ribotyping in 8 types occurring among the 1952-1956 strains and 10 types occurring among the 1992 strains. In 87% of the strains the results of the 2 typing methods were in accordance. However, 7 strains gave different results by the 2 methods including 2 strains with major differences. Penicillin resistance only occurred in a single genotype from the 1950's compared to 6 different genotypes among the 1992 strains. PMID- 7484551 TI - Variations with breed, age, season, yield, stage of lactation and herd in the concentration of urea in bulk milk and individual cow's milk. AB - The concentration of urea in the milk of 510 dairy cows in 10 herds was determined at regular intervals for a year. The herds contained approximately equal numbers of Swedish Red and White, and Swedish Holstein cows. The mean +/- sd concentration in the samples from individual cows was 5.32 +/- 1.13 mmol/l, and the mean concentration in bulk milk was 5.39 +/- 0.96 mmol/l. These values indicated that on average the herds were fed too much protein relative to their intake of energy throughout the year. Herd factors had a strong influence on the milk urea concentration. The concentration was lower during the first month of lactation than later in the lactation, and lower when the cows were housed during the winter than when they were grazing. There was a weak positive relationship between the daily milk yield and urea concentration, particularly during late lactation, but there was no relationship with either breed or age. Bulk milk urea was a reliable guide to the average urea concentration of a herd. PMID- 7484552 TI - D-dimer improves the prognostic value of combined clinical and laboratory data in equine gastrointestinal colic. AB - The discriminating ability of 15 parameters alone or in combinations, including results from analysis of plasma endotoxin, the Nycomed plasma D-Dimer test and phospholipase A2, were analyzed to predict morbidity and mortality in equine gastrointestinal colic. Endotoxaemia was a characteristic feature of the colic horses. The problem of adequately predicting nonsurvivors among colic horses required several parameters to be included in the logistic model: if the "classical parameters", (heart rate, respiratory rate, PCV, anion gap) were included in the model, addition of plasma D-dimer, phospholipase A2, and Cl- significantly improved the predictive value of the logistic model. Increasing heart rate and D-dimer together with decreasing chloride was a risk factor for nonsurvival. The sensitivity of this three-parameter logistic model to predict nonsurvival was 78% and specificity 77%. The Nycomed D-Dimer test is recommended as a horse-site test to predict disseminated intravascular coagulation and nonsurvival in equine colic. PMID- 7484553 TI - Lack of staphylococcal enterotoxin production among strains of Staphylococcus aureus from bovine mastitis in Denmark. PMID- 7484554 TI - Narthecium ossifragum associated photosensitization in sheep in the Faroe Islands. PMID- 7484555 TI - A case of Neospora associated bovine abortion in Sweden. PMID- 7484556 TI - Evaluating health services: the importance of patients' preferences and quality of life. AB - With limited resources available, we all would like to allocate health care dollars to do the most good. Clinical research tells us what outcomes to expect (in many cases) from the introduction of a health care program, a test, or a therapy. Even primitive cost analysis can assess the cost of such programs. The ability to place a value on health states is vital when assessing how patient outcomes influence the relative cost-effectiveness of medical procedures, therapies, and programs. Without a means to measure the value of a particular health state, one is left to compare apples with oranges and oranges with vacuum cleaners. In fact, comparisons of fruit and home appliances is relatively easy, because one can readily apply monetary values to apples, oranges, and vacuum cleaners and compare dollar amounts. How can one do the same for the outcomes of medical procedures and diagnostic tests? This is the challenge for health services and outcomes researchers throughout the world and, more urgently, the focus of policy makers, governments, and health insurers. The purpose of this paper is to describe quality-adjusted life-years (QALYs), a method that has successfully measured the outcomes of disparate health programs. We will introduce the QALY method, summarize the various methods of measuring and classifying health states, describe three methods that have been used to measure patients' preferences (utilities) for health states, and discuss the limitations of utility assessment and some controversies that result from the measurement and use of utilities and concerning health-related quality of life. Readers who are interested in general topics of radiology technology assessment and cost effectiveness analysis should consult other review articles [1-4]. PMID- 7484557 TI - Making difficult health policy decisions: most choices are not simply life or death. PMID- 7484558 TI - Outside financial interests in radiology offices: prevalence, payment, and practice patterns in involved offices. AB - OBJECTIVE: The purposes of our study were to (1) investigate the prevalence of different patterns of outside financial interests in radiology offices and of different methods of compensating these interests and (2) determine how the professional, operational, and practice characteristics of the offices vary with the patterns of outside interest. The term "office" means nonhospital practice sites, including those called imaging centers or clinics; "outside interests" refers to parties other than physicians in the group of physicians who serve the office. MATERIALS AND METHODS: Data are for 516 office sites served by radiology groups. Data were collected as part of a national, stratified random sample survey of radiology groups in the United States done by the American College of Radiology in 1992. Responses were weighted to make the results representative of all radiology groups in the United States with offices. RESULTS: Parties other than the physicians in the practice serving an office had a financial interest in 41% of offices. In about half of these (20% of all offices), physicians in the practice had no financial interest at all. The most common categories of outside financial interest were referring physicians (21%), hospitals (19%), and non physician entrepreneurs (12%); some offices reported more than one type of outside financial interest. All categories were more common in high-tech offices (offices that offered CT, MR imaging, or nuclear medicine studies) than in non high-tech offices. Overall, 50% of high-tech offices and 35% of non-high-tech offices had outside financial interests. The most common method of compensating outsiders was giving them shares or dividends or a percentage of the profits. The average number of procedures per full-time-equivalent radiologist was higher in offices in which only outsiders (and not the group serving the office) had a financial interest than in other offices, and fewer of the former offices offered mammography. Offices with no outside interests were more likely than those with any outsiders to regularly do chest, spinal, gastrointestinal, and urologic procedures. CONCLUSIONS: Outside financial interests, and the problems of cost and access they may engender, are fairly common in radiology offices. Nationally, joint ventures including referring physicians, which are now under legal attack, number more than 500. Differences in who owns radiology offices were associated with statistically significant variations among offices in workload, staffing, and types of procedures done. PMID- 7484559 TI - The sex ratio of American radiologists: comparison and implications by age, subspecialty, and type of practice. AB - OBJECTIVE: The purpose of this paper is to present results related to two questions regarding changes in the sex ratio of American radiologists. Do men and women in radiology have different patterns of subspecialization, postresidency training, board certification, or practice characteristics? Do differences in patterns between the sexes imply that the number of radiologists required in the future will change with a changing sex ratio of radiologists? MATERIALS AND METHODS: A survey questionnaire was mailed to a stratified random sample of 2804 radiologists, radiation oncologists, and nuclear medicine specialists drawn from the American Medical Association Physician Masterfile. The survey achieved a response rate of 69%. Stratification criteria included sex, age, and subspecialty. The survey questions included age, sex, subspecialty, training status, board certification, type of practice, principal work activity, source of income, hours worked, and amount of time away from the practice. Data analysis used descriptive statistics, ordinary least squares regression, and logit analysis. Weighting assured that results represent all radiologists. RESULTS: Only 13% of active radiologists who have finished training are women. The percentage varies with age; 6% of radiologists 45 years old or older; 22% of those 35-44 years old, and 23% of those younger than 35 years old were women. Differences in the sex ratio were not associated with differences in board certification or postresidency fellowships, but subspecialization differed by sex. Women were more likely than men to be salaried, to work part-time, to be engaged in teaching, and to work in an office rather than in a hospital. Differences in the sex ratio had little impact on estimates of the number of radiologists that will be needed in the future. CONCLUSION: Female radiologists have subspecialization and practice characteristics different from those of male radiologists. The increasing percentage of women in the profession will have little effect on the number of radiologists and radiation oncologists needed. PMID- 7484560 TI - Preoperative examination of lung transplant candidates: value of chest CT compared with chest radiography. AB - OBJECTIVE: The purpose of our study was to determine the usefulness of CT in examining candidates for lung transplantation to detect cancer not visible on plain chest radiographs (a finding that would exclude a patient from transplantation) and to determine which lung is more severely diseased to aid in the decision of which side to transplant. MATERIALS AND METHODS: We reviewed the chest radiographs and CT scans of 190 transplant candidates during a 3-year period for findings suggestive of neoplasm, including lung nodules and atelectasis; we also reviewed the symmetry of disease. In the latter category, patients with primary pulmonary hypertension, Eisenmenger's complex (pulmonary vascular, not parenchymal, disease), and cystic fibrosis (for which bilateral transplantation is performed) were excluded. A total of 190 plain chest radiographs, 180 thin-section CT scans, and 31 standard CT scans were reviewed retrospectively. RESULTS: Plain chest radiographs revealed 20 noncalcified nodules; 13 were 8 mm or more in diameter, and 7 were less than 8 mm. CT scans revealed 66 noncalcified nodules; 37 were 8 mm or more in diameter, and 29 were less than 8 mm. Eight non-calcified nodules seen on plain chest radiographs were either absent on CT scans (and follow-up plain chest radiographs) or appeared calcified on CT scans. Solitary nodules in three patients proved to be bronchogenic carcinomas; two of these lesions were identified only on CT scans. CT prompted a change in the determination of which lung was more severely diseased from that made on the basis of plain radiography for 27 of 169 patients (16%) evaluated. Of the 45 patients who subsequently underwent transplantation, CT prompted a change in the determination of which side to transplant from that made on the basis of plain radiography for 4 patients (9%). CONCLUSION: CT provides additional information to supplement plain radiography in the examination of lung transplant candidates. This information can alter patient management, particularly when bronchogenic carcinoma is detected, and enable a better determination of which lung is more severely diseased to aid in the decision of which lung to transplant for single-lung transplantation. PMID- 7484561 TI - CT of the bronchus intermedius: frequency and cause of a nodule in the posterior wall on normal scans. AB - OBJECTIVE: A focal nodule in the posterior wall of the bronchus intermedius is occasionally observed on normal CT scans. The purpose of this study was to determine the frequency and the anatomic cause of this finding. MATERIALS AND METHODS: We prospectively analyzed helical CT scans (10-mm collimation and 10 mm/sec table speed) for evaluating nodules in the posterior wall of the bronchus intermedius from 280 consecutive subjects. A focal round elevation in the posterior wall of the bronchus intermedius was considered a nodule. Frequency of visualization of the nodule was recorded. In patients showing such nodularity, additional enhanced CT scans with thinner slices (5-mm collimation and 5-mm/sec table speed) were obtained to elucidate the nature of the nodularity. RESULTS: Nodularity in the posterior wall of the bronchus intermedius was observed in 14/280 total subjects (5%) on 10-mm collimation CT scans. On thin-section CT scans, the nodularity was caused by a branch of the vein from the posterior segment of the right upper lobe (10/14, 71%) or by a branch of the vein from the superior segment of the right lower lobe (4/14, 29%). The vein from the upper lobe that caused nodularity in the posterior wall of the bronchus intermedius drained into either the inferior pulmonary vein (7/10) or the superior pulmonary vein (3/10). In all patients whose nodularity was caused by the vein from the superior segment of the right lower lobe, the vein drained into the inferior pulmonary vein. CONCLUSION: Focal nodularity in the posterior wall of the bronchus intermedius was caused by the draining pulmonary vein, and it was observed in 5% of normal subjects. This nodularity should be differentiated from uniform or lobular thickening associated with abnormality in the bronchus intermedius. PMID- 7484562 TI - Selenium-based digital radiography of the chest: radiologists' preference compared with film-screen radiographs. AB - OBJECTIVE: A new digital thoracic radiography system (Thoravision; Philips Medical Systems, Hamburg, Germany), which uses selenium as a detector material, was evaluated for observer preference. The system has been shown to have higher detection efficiency than conventional film-screen systems and thus could provide an image with reduced noise. The hypothesis tested in this study was that the selenium-based digital system would provide an image appearance for conventional thoracic imaging that would be equal or superior to that provided by a conventional film-screen system. MATERIALS AND METHODS: Fifty-three patient volunteers were imaged at 120 kV with both the selenium-based system and a thoracic film-screen combination system (InSight HC; Kodak, Rochester, NY). Posteroanterior and lateral images were acquired with both systems, for a total of 212 images. Both imaging systems included a stationary 12:1 antiscatter grid. Exposures were the same for both imaging systems, and the digital images were printed to film. Images for the same patient were compared by six observers- three specialized chest radiologists and three general radiologists. Images included both normal chest radiographs and radiographs with abnormal findings. Each pair of images was ranked on a scale from 1 to 5 for preference of technique, with a score of 3 indicating no preference. Eleven anatomic features were evaluated in the posteroanterior views, and six features were evaluated in the lateral views. Statistical significance of preference was evaluated with Student's t test. RESULTS: The chest radiologists had a statistically significant preference for the selenium-based system for all 17 features (p < .001). The general radiologists had a statistically significant preference for the selenium based system for visualization of 10 of the 17 features (p < .05). Neither group had a statistically significant preference for the conventional images in any category. CONCLUSION: The selenium-based system provided an image appearance that was significantly preferred by all radiologists, more strongly by those specializing in chest radiography. This study demonstrates that a digital thoracic imaging system can routinely produce images that are perceived as equal or superior to conventional images. PMID- 7484565 TI - Problems with the American College of Radiology Standard for Diagnostic Mammography. AB - In an effort to help assure quality, the American College of Radiology (ACR) has developed standards for many imaging procedures. One of the most recent standards adopted is the ACR Standard for Diagnostic Mammography and Problem-Solving Breast Evaluation. Despite the fact that this Standard apparently went through a lengthy review process (although this author, a member of the Breast Cancer Task Force of the ACR, never gave his approval), it contains many unsupportable recommendations. PMID- 7484563 TI - Pulmonary thromboembolism: spectrum of findings on CT. AB - CT has emerged as a potentially important diagnostic technique in pulmonary thromboembolism that can provide direct visualization of the obstructing embolus and its associated vascular and pleuroparenchymal sequelae. Although the role of contrast-enhanced CT in the diagnostic algorithm of pulmonary embolism has not been defined, patients at risk for pulmonary embolism frequently undergo thoracic CT as part of the assessment for other cardiopulmonary conditions. As such, CT may provide the first indication of clinically significant pulmonary embolic disease. This essay reviews the vascular, pleural, and parenchymal features of pulmonary thromboembolism observed on CT scans, describes the limitations of CT, and discusses the imaging features of pulmonary embolism that overlap other clinical conditions. PMID- 7484564 TI - Bronchopleural-subarachnoid fistula manifesting as intracranial gas on CT scans. PMID- 7484566 TI - Re: Problems with the American College of Radiology Standard for Diagnostic Mammography. PMID- 7484567 TI - Dr. Charles Richard Drew. PMID- 7484568 TI - Screening mammography in community practice: positive predictive value of abnormal findings and yield of follow-up diagnostic procedures. AB - OBJECTIVE: The purpose of this study was to gather from 50 community mammography practices that were included in the National Survey of Mammography Facilities data concerning abnormal findings on screening mammograms to determine the frequency of various recommendations made for patients who had abnormal findings and to compare these recommendations with the frequency with which the procedures were actually performed. We also determined the positive predictive value of screening mammograms (the number of cancers detected per 100 abnormal findings) and the yield (the number of cancers detected per 100 procedures done) of various diagnostic procedures done because of abnormal findings. MATERIALS AND METHODS: We identified 1717 screening mammograms done in the last half of 1991 that had abnormal findings. Radiologic recommendations and follow-up procedures, including repeat standard (screening) mammography, additional mammographic views, sonography, clinical breast examination, needle aspiration, needle biopsy, and open biopsy, were identified for all of the cases from the radiologic records, and follow-up data were obtained from referring physicians. The positive predictive value and yield in the National Survey of Mammography Facilities were compared with data from the mammography screening practice of the University of California at San Francisco (UCSF), a facility noted for its clinical efficiency. RESULTS: We estimate that 11% of all screening mammograms resulted in a recommendation for further diagnostic procedures. These 1717 mammograms with abnormal findings led to the following recommendations and procedures: repeat standard (screening) mammography, 610 (recommended)/635 (performed); additional mammographic views, 785/707; sonography, 400/345; biopsy, 189/229; and needle aspiration, 21/51. More procedures were done than were recommended in some cases because the results of certain procedures often led to the performance of other, additional procedures. The positive predictive value for screening examinations with abnormal findings was 3.5%, and the yield for open biopsy was 21%. In the UCSF data base, the positive predictive value for examinations with abnormal findings was 10%, and the yield for open biopsy was 34%. CONCLUSION: The positive predictive value for examinations with abnormal findings and the yield for diagnostic procedures performed as a result of abnormal findings in 50 community radiologic facilities were higher than those reported in some earlier studies, a fact that raised concern about the induced cost of screening mammography. However, these values were low compared with those in the UCSF data base. This fact was particularly true of repeat standard (screening) mammography. PMID- 7484569 TI - Surgical removal of ruptured breast implants: the use of intraoperative sonography in localizing free silicone. PMID- 7484571 TI - Pneumatosis intestinalis in adults with AIDS: clinical significance and imaging findings. AB - OBJECTIVE: The significance of pneumatosis intestinalis in adults with AIDS is unknown. Adults with AIDS are predisposed to pneumatosis intestinalis. The purpose of this study is to determine the clinical significance and to characterize the imaging appearance of this finding in patients with AIDS. MATERIALS AND METHODS: In just over 3 years at our institution, pneumatosis intestinalis in adults with AIDS was detected in six cases. Medical records of those cases were retrospectively analyzed for clinical history, laboratory data, and surgical and pathologic findings. CT scans and abdominal radiographs were analyzed for location and severity of pneumatosis, presence of portal venous gas, pneumoperitoneum, and cystic versus linear gas. RESULTS: Five of the six patients with AIDS-associated pneumatosis intestinalis were managed conservatively or medically without surgical procedures and were discharged from the hospital with resolved or decreasing gastrointestinal complaints. The other patient underwent an exploratory laparotomy because of free intraperitoneal and retroperitoneal air; however, no bowel perforation was found at surgery. CONCLUSION: Pneumatosis intestinalis is a late-stage phenomenon in adult patients with AIDS that characteristically involves the cecum or right colon. Pneumatosis in patients with AIDS may be an indolent abnormality and does not necessarily constitute a surgical emergency or a sign of impending bowel necrosis. PMID- 7484570 TI - Intramural pseudodiverticulosis of the esophagus detected on barium esophagograms: increased prevalence in patients with esophageal carcinoma. AB - OBJECTIVE: We tested the hypothesis that intramural pseudodiverticulosis of the esophagus is more prevalent in patients with esophageal carcinoma than in randomly selected patients who undergo esophagography for other indications. Such an association would prompt a careful search for carcinoma after esophageal intramural pseudodiverticulosis is found. MATERIALS AND METHODS: Single- and double-contrast esophagograms of 245 patients with esophageal carcinoma were retrospectively reviewed. A control group of 6400 esophagograms obtained for indications other than esophageal carcinoma was also reviewed. The statistical significance of the difference in prevalence of esophageal intramural pseudodiverticula between patients with esophageal carcinoma and the control group was tested using the chi-square test. The significance of difference between the number of esophageal intramural pseudodiverticula in patients with esophageal dilatation and the number in those without dilatation was tested using a small-sample nonparametric test. RESULTS: Intramural pseudodiverticulosis of the esophagus was found in 11 patients with esophageal carcinoma (4.5%) and in six control subjects (0.09%). Intramural pseudodiverticulosis of the esophagus was present opposite the tumor as well as both proximally and distally. The number of cases of intramural diverticulosis in patients with esophageal carcinoma and dilatation was not significantly higher than that in patients with carcinoma but without dilatation (p > .1) CONCLUSION: The prevalence of esophageal intramural pseudodiverticulosis is significantly higher in patients with esophageal carcinoma than in patients who underwent esophagography for other indications (p < .0002). This association implies increased risk of esophageal carcinoma in patients with intramural pseudodiverticulosis. Periodic surveillance of patients with intramural pseudodiverticulosis of the esophagus for esophageal carcinoma may be worthwhile. PMID- 7484573 TI - MR cholangiopancreatography: findings on 3D fast spin-echo imaging. PMID- 7484572 TI - Diagnosis of abdominal tuberculosis: sonographic findings in patients with early disease. AB - OBJECTIVE: The diagnosis of abdominal tuberculosis is often difficult, because clinical manifestations and results of laboratory studies are nonspecific. If sonographic findings are sufficiently characteristic for diagnosis, sonography would be useful, especially in India, where abdominal tuberculosis is common and more expensive imaging techniques are not easily available. Accordingly, we performed sonography to establish the sonographic findings in cases of early tuberculosis in 56 patients with abdominal tuberculosis who had normal barium studies of the small bowel. SUBJECTS AND METHODS: Fifty-six patients with clinical features suggestive of abdominal tuberculosis (history of fever, abdominal pain, and weight loss) with no history of intestinal obstruction and normal barium studies of the small bowel had abdominal sonography. All sonograms were independently assessed by three radiologists, and the findings were tabulated by consensus. Diagnosis of tuberculosis was confirmed by sonographically guided biopsy of mesenteric lymph nodes in 19 patients, analysis of aspirated ascitic fluid in 12, and response to antituberculous chemotherapy in 25. Sonography was repeated 1, 3, 6, and 12 months after antituberculous chemotherapy was begun. Abdominal sonograms were also performed in 30 healthy volunteers, and measurements of mesenteric thickness were recorded. The mesenteric thickness was statistically compared in two groups of patients: patients at presentation with patients at the end of antituberculous chemotherapy and patients at presentation with healthy individuals. RESULTS: The mesenteric thickness in healthy individuals ranged from 5 to 14 mm. Sonographic findings in all patients with abdominal tuberculosis included an echogenic thickened mesentery (> or = 15 mm) with mesenteric lymphadenopathy. Other findings were dilated small bowel loops in 38 patients, minimal ascites in 17, matted small bowel loops in five, and omental thickening with altered echogenicity in three. Regression of these changes was noted on follow-up of all patients undergoing treatment. CONCLUSION: The characteristic sonographic features of early abdominal tuberculosis are mesenteric thickness of 15 mm or more and an increase in the mesenteric echogenicity (due to fat deposition), combined with mesenteric lymphadenopathy. Presence of dilated small bowel loops and ascites further substantiate the diagnosis. PMID- 7484574 TI - Single-pass CT of hepatic tumors: value of globular enhancement in distinguishing hemangiomas from hypervascular metastases. AB - OBJECTIVE: The purpose of this study was to evaluate the sensitivity and specificity of globular enhancement for differentiating hepatic hemangiomas from hypervascular metastases on single-pass, contrast-enhanced CT scans. Globular enhancement was defined as enhancing nodules less than 1 cm seen within a lesion. MATERIALS AND METHODS: Fifty CT examinations were retrospectively evaluated in a blinded fashion by two independent reviewers. The CT studies were done with nonhelical technique after the IV injection of 150 ml of contrast material. The series included 25 patients with histologically proven hypervascular hepatic metastases (carcinoid, islet cell carcinoma, and leiomyosarcoma) and 25 patients with clinically proven hepatic hemangiomas. Patients with hemangiomas were clinically stable for at least 2 years after the CT studies. A single lesion was isolated from the first-pass, contrast-enhanced portion of each examination; the remainder of the examination was excluded from the review to minimize reviewer bias. Each lesion was evaluated for (1) the presence or absence of globular enhancement (defined as enhancing nodules less than 1 cm seen within a lesion), (2) the density of globular enhancement relative to that of the aorta, (3) the degree of border definition (well or poorly marginated), and (4) the presence or absence of a hypodense halo. A diagnostic impression was then recorded for each lesion. RESULTS: Globular enhancement was 88% sensitive and 84-100% specific for differentiating hepatic hemangiomas from hypervascular metastases (p < .001). A mean of 62% of hemangiomas showed globular enhancement isodense relative to that of the aorta; none of the metastases showed globular, isodense enhancement. The majority of the metastases showed nonglobular enhancement (mean, 92%). The reviewers showed 84% agreement in the identification of (1) globular enhancement in hemangiomas, (2) lack of globular enhancement in metastases, and (3) globular enhancement in the combined set of all lesions. Neither the presence of a hypodense halo nor the degree of border definition was significant in distinguishing between the two groups of lesions. The reviewers showed 96% agreement in the categorization of metastases and 76% agreement in the categorization of hemangiomas. There was 86% agreement in the categorization of all lesions. Overall, reviewers diagnosed a mean of 89% of lesions correctly. A mean of 98% of metastases and a mean of 80% of hemangiomas were diagnosed correctly. CONCLUSIONS: Globular enhancement is highly sensitive (88%) and specific (84-100%) for differentiating hepatic hemangiomas from hypervascular metastases on single-pass, contrast-enhanced CT scans. PMID- 7484575 TI - MR diagnosis of hepatic metastases from neuroendocrine tumors versus hemangiomas: relative merits of dynamic gadolinium chelate-enhanced gradient-recalled echo and unenhanced spin-echo images. AB - OBJECTIVE: Hepatic metastases from neuroendocrine tumors are often markedly hyperintense on unenhanced T2-weighted MR images, making their appearance similar to that of cavernous hemangiomas. In contrast, cavernous hemangiomas show characteristic enhancement on dynamic gadolinium chelate-enhanced gradient recalled echo MR images. The purpose of this study was to determine the relative merits of dynamic gadolinium chelate-enhanced gradient-recalled echo MR imaging versus MR imaging with unenhanced spin-echo pulse sequences for distinguishing between hepatic metastases from neuroendocrine tumors and cavernous hemangiomas. MATERIALS AND METHODS: The unenhanced spin-echo and dynamic gradient-recalled echo MR images obtained after IV administration of a gadolinium chelate in 28 patients (14 patients with pathologically proven hepatic metastases from neuroendocrine tumors and 14 patients with hepatic cavernous hemangiomas) were reviewed blindly and independently by three interpreters. Unenhanced spin-echo and dynamic gadolinium chelate-enhanced gradient-recalled echo MR images were compared for accuracy in characterizing liver lesions. RESULTS: The most intense enhancement of hepatic metastases from neuroendocrine tumors was observed on early dynamic gadolinium chelate-enhanced gradient-recalled echo MR images; enhancement was peripheral in four patients, global and heterogeneous in seven patients, and global and homogeneous in three patients. On late dynamic gadolinium chelate-enhanced gradient-recalled echo MR images, enhancement of hepatic metastases from neuroendocrine tumors was predominantly peripheral in five patients, global and heterogeneous in five patients, and global and homogeneous in four patients. Differentiation between cavernous hemangiomas and hepatic metastases from neuroendocrine tumors was impossible in five cases with unenhanced spin-echo MR imaging alone, in five cases with dynamic gadolinium chelate-enhanced gradient-recalled echo MR imaging alone, and in no case with the combination of unenhanced spin-echo MR imaging and dynamic gadolinium chelate enhanced gradient-recalled echo MR imaging. In comparison with unenhanced spin echo MR imaging alone or dynamic gadolinium chelate-enhanced gradient-recalled echo MR imaging alone, the combination of unenhanced spin-echo MR imaging and dynamic gadolinium chelate-enhanced gradient-recalled echo MR imaging allowed significantly (p < .001) clearer differentiation between hepatic metastases from neuroendocrine tumors and cavernous hemangiomas. CONCLUSIONS: Early enhancement and heterogeneity on dynamic gadolinium chelate-enhanced gradient-recalled echo MR images are the most common features of hepatic metastases from neuroendocrine tumors. The combination of unenhanced spin-echo and dynamic gadolinium chelate enhanced gradient-recalled echo MR images allows more accurate characterization of hepatic metastases from neuroendocrine tumors and clearer differentiation from cavernous hemangiomas. PMID- 7484576 TI - Hepatic arteriography in patients with hepatocellular carcinoma: change in findings caused by balloon occlusion of tumor-draining hepatic veins. AB - OBJECTIVE: A previous study suggested that temporary occlusion of a segment of the hepatic vein causes an increase in arterial flow and retrograde portal flow in the occluded segment. Such occlusions might improve the efficacy of arterial infusion therapy. Accordingly, we studied the change in blood flow visible on hepatic arteriograms when a segment of the hepatic vein is temporarily occluded in patients with hepatocellular carcinoma. MATERIALS AND METHODS: The study group consisted of 24 patients with nodular-type hepatocellular carcinoma. Conventional hepatic arteriography was followed by hepatic arteriography performed using a balloon catheter to occlude the hepatic vein that was most closely associated with the tumor. Visualization of the tumor-draining veins, portal vein branches, the degree of tumor vascularity, and the density of the hepatogram on the hepatic arteriogram were retrospectively compared before and during venous occlusion. The veins were evaluated visually; an increase of tumor vascularity was defined as an increase in the number of countable tumor vessels during occlusion, and a dense hepatogram was considered to be a definite sinusoidgram induced by venous occlusion. RESULTS: Conventional hepatic arteriography showed the tumor-draining veins to be branches of the portal vein in only two of the 24 patients (8%). Hepatic arteriography during venous occlusion, however, showed the tumor-draining veins to be branches of the portal vein in four of the patients (17%). An increase in the degree of tumor vascularity with venous occlusion was observed only in a patient with an initial arteriohepatic vein shunt. Dense hepatogram and hepatofugal opacification of the portal vein branches in the occluded, tumor bearing segment were obtained in 10 patients (42%). Eight of these did not have liver cirrhosis, whereas all of the remaining 14 patients did (p < .001). CONCLUSION: Our results suggest that occlusion of a segment of the hepatic vein may be useful during arterial infusion of hepatocellular carcinoma. PMID- 7484577 TI - Hepatic involvement in Osler-Weber-Rendu disease: findings on pulsed and color Doppler sonography. AB - OBJECTIVE: The purpose of this study was to compare the hemodynamics of the liver vasculature as determined by Doppler sonography for patients with Osler-Weber Rendu disease and for healthy subjects. SUBJECTS AND METHODS: Real-time color Doppler sonography and pulsed Doppler sonography were used to study 10 patients with Osler-Weber-Rendu disease and 25 healthy subjects. Vessel diameter and flow velocity in the hepatic artery and its branches as well as in the portal vein and its segmental branches were determined. Flow patterns in the hepatic veins were analyzed. The clinical manifestations of the disease were correlated with the Doppler sonographic findings. RESULTS: The hepatic artery and its branches were dilated and tortuous, and flow velocity was greater in patients than in healthy subjects; hepatic artery velocities (mean +/- SD) were 153 +/- 65.2 cm/sec versus 64.9 +/- 11.4 cm/sec (p < .005). The resistive index measured in the hepatic artery did not differ significantly between the two groups (p was not significant). No intrahepatic arteriovenous shunt sites were detected by color Doppler sonography. However, arterioportal shunts were observed in two patients, one of whom also had a portovenous shunt. A portovenous shunt and multiple shunts between hepatic veins were found in another patient. Portal venous flow was similar in both groups. No relationship between Doppler sonographic changes and clinical manifestations was found. CONCLUSION: The multiple microscopic arteriovenous shunts found in Osler-Weber-Rendu disease were not detected in our patients but resulted in dilatation and increased flow velocity in the hepatic artery. Flow velocity in the portal vein appeared to be undisturbed. Large intrahepatic shunts were easily outlined by Doppler sonography. PMID- 7484578 TI - Hepatocellular adenoma. PMID- 7484580 TI - Imaging of intrahepatic cholangiocarcinoma: 2. Hilar cholangiocarcinoma. AB - Hilar cholangiocarcinoma (also called Klatskin's tumor) is more common than peripheral cholangiocarcinoma. Sonography, CT, MR imaging, angiography, and cholangiography can suggest the diagnosis, but the major issue of imaging with this tumor is to determine whether the tumor is resectable. The anatomic location of hilar cholangiocarcinoma makes resection difficult, so that surgical exploration of patients with this condition should be undertaken only when preoperative evaluation has shown a potential for curative resection. Preoperative assessment of resectability of hilar cholangiocarcinoma is often extensive, requiring several types of imaging. This pictorial essay reviews the imaging features of hilar cholangiocarcinoma. The role of imaging in the preoperative planning, with specific emphasis on staging extent of disease, including hepatic and vascular involvement, is discussed and illustrated. PMID- 7484581 TI - Helical CT for the preoperative localization of islet cell tumors of the pancreas: value of arterial and parenchymal phase images. AB - OBJECTIVE: In two-phase helical CT, organs are scanned in both the arterial and the parenchymal phase after IV injection of contrast material. The purpose of our study was to evaluate the usefulness of two-phase helical CT for the preoperative localization of pancreatic islet cell tumors. MATERIALS AND METHODS: Ten patients with 11 surgically proven islet cell tumors were included in the study. Helical CT scans of the pancreas were obtained with 5-mm collimation and overlapping image reconstruction. Images were obtained in the arterial and parenchymal phases after contrast material injection. A test bolus was used for tailored bolus timing. Images were analyzed by two observers. If a tumor was detected, its density relative to the surrounding parenchyma was scored by consensus, using a five-point scale. RESULTS: Nine of 11 tumors could be located using two-phase helical CT (sensitivity, 82%), including one 4-mm gastrinoma. Two lesions smaller than 5 mm could not be visualized. Two tumors were easier to detect (n = 1) or could only be detected on parenchymal-phase images (n = 1). Tumor conspicuity was better on arterial-phase images in two patients. CONCLUSION: Our results show that obtaining CT scans in both the arterial and the parenchymal phase leads to improved detection of pancreatic islet cell tumors. PMID- 7484579 TI - Imaging of intrahepatic cholangiocarcinoma: 1. Peripheral cholangiocarcinoma. AB - Cholangiocarcinoma is the second most common primary hepatic malignant tumor after hepatocellular carcinoma, accounting for 5-30% of all primary hepatic malignant tumors [1]. Intrahepatic cholangiocarcinomas can be classified as peripheral cholangiocarcinoma, which originates from an interlobular biliary duct, or as hilar cholangiocarcinoma, which originates from a main hepatic duct or from the bifurcation of the common hepatic duct. Intrahepatic cholangiocarcinomas account for only about half of cholangiocarcinomas, and this pictorial essay focuses only on the peripheral form of the disease. Clinically, therapeutically, and radiologically, these two types of cholangiocarcinomas differ. Features suggestive of the diagnosis of peripheral cholangiocarcinoma can be shown by sonography, CT, and MR imaging. Cholangiography and angiography have a limited role in evaluating this neoplasm that manifests as a focal mass. This essay reviews the appearances of peripheral cholangiocarcinoma and discusses the various imaging techniques that can be used to evaluate this unusual tumor that is often resectable and potentially curable. PMID- 7484582 TI - Diagnosis of renal artery stenosis in transplanted kidneys: value of Doppler waveform analysis of the intrarenal arteries. AB - OBJECTIVE: The objective of this study was to determine in renal transplant patients if the acceleration time and subjective assessment of dampening of the waveforms from the intrarenal arteries improves the accuracy of detecting a hemodynamically significant (> or = 50%) proximal arterial stenosis compared with measurements of peak systolic velocity from a main renal artery. MATERIALS AND METHODS: In 15 patients, the findings of 19 Doppler sonograms and corresponding arteriograms of their renal transplants were reviewed, with arteriography serving as the gold standard. Four patients had a significant proximal arterial stenosis; three were of the main renal artery and one was of the adjacent external iliac artery proximal to the anastomosis with the renal artery. RESULTS: We found a significant prolongation of the acceleration time in patients with a significant proximal arterial stenosis (p = .0004). Use of a threshold acceleration time of 0.10 sec or subjective assessment of dampening of the waveforms resulted in an accuracy of 95% in detecting a significant proximal arterial stenosis. This compared with an accuracy of 62% in detecting a significant proximal arterial stenosis using a peak systolic velocity threshold of 2.0 m/sec as the sole criterion. Using intrarenal arterial Doppler waveform parameters alone would have spared arteriography in 11 patients and would have detected three of four significant proximal arterial stenoses. CONCLUSION: In this study, Doppler waveform analysis of the intrarenal arteries improved the accuracy of screening for a significant proximal arterial stenosis. The results suggest that such analyses can be used to spare many patients with suspected renal vascular hypertension from unnecessary arteriography. PMID- 7484583 TI - Diagnosis of renal artery stenosis: feasibility of combining MR angiography, MR renography, and gadopentetate-based measurements of glomerular filtration rate. AB - OBJECTIVE: Our aim was to evaluate the feasibility of combining in a single test (1) structural evaluation of renal arteries with MR angiography, (2) functional evaluation of global glomerular filtration rates calculated on the basis of plasma disappearance of gadopentetate dimeglumine, and (3) renographic analysis of individual kidneys based on the dynamic changes in signal intensity that occur after administration of gadopentetate dimeglumine. SUBJECTS AND METHODS: We used unenhanced MR angiography to measure patency of the renal arteries in 10 healthy volunteers and in 10 patients with renal artery stenosis. Calculations of global glomerular filtration rate were based on measurements of plasma disappearance of gadopentetate dimeglumine as shown by MR relaxometry. For renography with gadopentetate dimeglumine, we generated curves that showed changes in signal intensity in both kidneys over time; intrarenal kinetics were studied by measuring the time of arrival of gadopentetate dimeglumine in the cortex and outer medulla of the kidney. Conventional angiograms, measurements of global glomerular filtration rate based on plasma disappearance of 99mTc-DTPA, and 99mTc DTPA renograms were used as reference standards. We compared the two different methods of determining global glomerular filtration rates by computing the correlation coefficient of the linear regression of rates derived from studies with gadopentetate dimeglumine versus rates derived from studies with 99mTc-DTPA. RESULTS: In all volunteers, renal arteries were well visualized, and global glomerular filtration rates based on plasma clearance of gadopentetate dimeglumine were normal. In nine of 10 patients, correlation was good between findings on MR angiograms and findings on conventional arteriograms. Finding were discordant in one patient because the patient moved during the MR angiography. For all six patients studied, correlation was good between measurements of global glomerular filtration rates based on plasma clearance of gadopentetate dimeglumine and those based on clearance of 99mTc-DTPA (r = 98%). CONCLUSION: Our results suggest the potential of magnetic resonance for a comprehensive approach for detection of renal artery stenosis. This novel approach provides structural evaluation of renal arteries with unenhanced MR angiography. MR renography is done and global glomerular filtration rates are determined by using MR relaxometry after injection of contrast material. Corticomedullary transit times can be determined on the basis of the dynamic changes in signal intensity that occur after administration of gadopentetate dimeglumine. PMID- 7484584 TI - Uterine rupture with fetal death following blunt trauma. PMID- 7484585 TI - An imaging algorithm for the differential diagnosis of adrenal adenomas and metastases. AB - OBJECTIVE: The purpose of this study was to develop an algorithm using CT and chemical-shift MR imaging for the characterization of adrenal masses in patients with a primary cancer and no other evidence of metastatic disease. SUBJECTS AND METHODS: Thirty-three patients with 37 adrenal masses (19 metastases, 18 adenomas), all of whom had a known primary cancer, were studied with noncontrast CT and chemical-shift MR imaging (1.5 T). Lesion size and density in Hounsfield units (H) were determined by CT. Adrenal signal intensity normalized to that of spleen was used to calculate adrenal-spleen ratio (ASR), defined as the percentage of signal remaining in the opposed-phase image relative to the in phase image. Lesions less than or equal to 0 H were classified as benign, lesions greater than 20 H were regarded as malignant, and lesions between 0 and 20 H were regarded as indeterminate. Diagnoses were confirmed by biopsy (for 19 lesions) or by follow-up imaging (for 18 lesions). An imaging algorithm was derived by determining the relative value of CT and MR imaging for diagnosing the lesions. The reimbursement rates for CT-guided biopsy and MR imaging of the abdomen were obtained from Medicare. RESULTS: All 13 lesions of 0 or less H were correctly classified as benign by CT. ASR was less than 70 in 10 of these 13. In another 13 lesions, H was greater than 20; all were malignant and all had an ASR greater than 80. Of 11 CT-indeterminate lesions, four of five adenomas had an ASR less than 70, and four of six metastases had an ASR greater than 80. Two malignant lesions had ASRs between 70 and 80 and were diagnosed by biopsy findings. One CT indeterminate adenoma had an ASR of 84 and was diagnosed by biopsy findings. The reimbursement rate by Medicare is similar for CT-guided biopsy with pathologic interpretation and for MR imaging of the abdomen. CONCLUSION: An algorithm was developed for diagnosis of adrenal lesions that uses the density reading on noncontrast CT as the first step, with chemical-shift MR imaging for CT indeterminate lesions. In this algorithm, lesions of 0 H or less may be regarded as benign and further work-up is not required. Lesions with a density greater than 20 H are likely malignant and should be biopsied when the result will influence management. For CT-indeterminate lesions, we recommend chemical-shift MR imaging. An ASR threshold of 70 indicates a benign lesion, and no further workup is required in these patients. Lesions with an ASR greater than 70 should have a biopsy performed, depending on the clinical situation. The above algorithm is cost-effective and reduces the number of biopsies required without reducing the sensitivity of detecting malignant lesions. PMID- 7484586 TI - Adrenal carcinoma in a patient with Gardner's syndrome: imaging findings. PMID- 7484587 TI - Ejaculatory duct obstruction: diagnosis and treatment with transrectal sonography. AB - Ejaculatory duct obstruction is thought to be a relatively uncommon but readily treatable cause of male infertility. Obstruction is frequently caused by cysts originating from or impinging on the ejaculatory ducts. The use of transrectal sonography has facilitated identification of patients with ejaculatory duct obstruction and has led to the realization that this abnormality is more common than was previously recognized. Transrectal sonography now plays a critical role in both the diagnosis and the treatment of this condition. The purpose of this pictorial essay is to illustrate the use of transrectal sonography in the diagnosis and treatment of ejaculatory duct obstruction. PMID- 7484588 TI - Condyloma acuminatum of the bladder: radiologic findings. PMID- 7484589 TI - Cost-effectiveness of MR imaging in evaluating polymyositis. AB - OBJECTIVE: Polymyositis is an inflammatory disorder of muscle that is often difficult to diagnose. The purpose of this study was to assess the usefulness and cost-effectiveness of MR imaging as a biopsy guide in patients with suspected polymyositis as an alternative to a blind biopsy. SUBJECTS AND METHODS: Our subjects were 25 patients with clinically suspected and ultimately confirmed polymyositis, 14 of whom had preoperative MR imaging at 1.5 T (including axial T1 weighted, axial T2-weighted [fat saturated], coronal T1-weighted, and coronal fast short-T1 inversion recovery images of both entire extremities). Images were prospectively interpreted for a specific location for a biopsy that showed the most muscle edema with the least atrophy. Surgical results and the estimated true cost of medical care (cost-accounting method) were compared between patients with preoperative MR images and patients whose biopsy site was selected on the basis of clinical criteria. RESULTS: Among the 14 patients whose biopsy site was selected on the basis of MR findings, only one biopsy result was false-negative. Patients without imaging had five biopsies with false-negative results. Patients with preoperative MR images averaged actual medical costs of $14,000 compared with $20,000 for those without MR images. Hospitalization time averaged 12 days for patients with MR images for guidance compared with 14 days for those without MR images. The average daily cost was $1,150 for the patients with MR images compared with $1,390 for those without. CONCLUSION: We conclude that MR imaging not only can aid in surgical planning but also can decrease the costs of patient care for patients with suspected polymyositis. PMID- 7484590 TI - Endovascular repair of abdominal aortic aneurysm: value of postoperative follow up with helical CT. AB - OBJECTIVE: Transfemorally placed endoluminal grafts are currently being evaluated as an alternative to open surgery for the treatment of abdominal aortic aneurysms. We determined the value of helical CT for the follow-up of patients treated with this new procedure. The purposes of this study were to determine CT features of a technically successful procedure, detect complications, and compare findings on CT scans obtained 24-48 hr after insertion of the graft with findings on angiograms obtained at the end of the endovascular procedure. SUBJECTS AND METHODS: Seven patients with large abdominal aortic aneurysms had helical CT within 48 hr after transfemoral insertion of an endoluminal graft. Findings on these CT scans were compared with findings on digital completion angiograms obtained immediately after placement of the graft. Additional follow-up CT scans were obtained for up to 15 months (mean, 8 months). The size of the aneurysmal sac; completeness of perigraft thrombosis; and position, shape, and patency of the device were recorded. RESULTS: CT scans obtained 24-48 hr after placement of the grafts showed complete thrombosis of the aneurysmal sac in three patients and incomplete thrombosis with patent perigraft channels in the four remaining patients. Angiograms showed a patent perigraft channel in only one patient. Two of four initially patent channels subsequently closed, but one of them recurred. Of four thrombosed aneurysms, two decreased in size, and two were unchanged on later follow-up. Of three aneurysms associated with perigraft channels, two became enlarged and one was stable. On the basis of CT criteria, successful endovascular repair was shown in four (57%) of seven patients. CONCLUSION: Helical CT is a sensitive means of evaluating the efficacy of endoluminal grafts. Decreased or stable size of the aneurysmal sac without perigraft channels on late follow-up CT signifies technical success. Persistence or recurrence of perigraft channels is the most likely cause of later enlargement of an aneurysm and therefore suggests procedural failure. Helical CT was more sensitive than angiography for detection of perigraft channels that occurred soon after treatment. PMID- 7484591 TI - Inflammatory aneurysms of the abdominal aorta: CT findings. AB - Inflammatory aneurysm of the abdominal aorta (IAAA) is a variant of atherosclerotic aneurysm that is characterized by inflammatory and/or fibrotic changes in the periaortic regions of the retroperitoneum [1, 2]. These inflammatory and/or fibrotic changes are probably the result of a local autoallergic reaction to certain components of atherosclerotic plaques [2]. This distinct entity has important implications, as the periaortic fibrotic tissue adherent to ureters, the duodenum, and the inferior vena cava may complicate surgical repair [1, 2]. The purpose of this essay is to illustrate the CT appearance of IAAA, with emphasis on the identification of IAAA and differentiation from conventional aortic aneurysms, evaluation of the involvement of adjacent structures by the periaortic fibrosis, and evaluation of the retroperitoneum after aneurysmal repair to analyze the resolution or the persistence of the periaortic fibrosis. PMID- 7484592 TI - Use of a perfusion balloon angioplasty catheter during femoral artery angioplasty. PMID- 7484593 TI - Plastic bag delivery system for hand injection of carbon dioxide. PMID- 7484594 TI - The ventriculus terminalis of the spinal cord in the neonate: a normal variant on sonography. AB - OBJECTIVE: The ventriculus terminalis, or fifth ventricle, is an ependymal-lined residual lumen of the caudal most portion of the spinal cord (the conus medullaris). We describe five neonates in whom this normal variant was depicted on sonography. MATERIALS AND METHODS: Screening lumbosacral spinal sonograms are performed at our institutions on neonates with lumbosacral cutaneous stigmata. Over a 2-year period (1993-1995), we prospectively found five cases in which spinal sonograms showed the ventriculus terminalis of the conus medullaris in term neonates. Two of the five received follow-up spinal sonograms at the age of 3 months. RESULTS: Sonograms of the lumbosacral spine of these five neonates showed an unusual hypoechoic or split appearance of the central echo complex of the conus medullaris. No other abnormalities were noted, and the finding was unchanged on the follow-up sonograms performed on two of the infants at the age of 3 months. These infants have remained asymptomatic during clinical follow-up periods of up to 2 years. CONCLUSION: The ventriculus terminalis is a normal anatomic variant of the conus medullaris that can be visualized on spinal sonograms in neonates. PMID- 7484595 TI - Scrotal inguinal hernia in a fetus: sonographic diagnosis. PMID- 7484596 TI - Detection and assessment of intracranial aneurysms: value of CT angiography with shaded-surface display. AB - OBJECTIVE: The purpose of this study was to evaluate the efficacy of circle of Willis CT angiography with shaded-surface display in the detection and assessment of suspected intracranial aneurysms. MATERIALS AND METHODS: Twenty-three patients who had clinical or radiologic suspicion of intracranial aneurysms were studied prospectively with CT angiography and conventional angiography. The images were obtained and interpreted in a double-blind fashion by different radiologists. Conventional angiography, which was the reference standard in this study, diagnosed 17 aneurysms in 15 patients. The usefulness of the two types of images for surgical planning was evaluated by two neurosurgeons. RESULTS: No aneurysms were detected with either technique in eight patients. Both techniques showed 15 aneurysms in 14 patients. The maximum dimensions were less than 3.0 mm in three cases, 3.0-5.0 mm in four cases, and greater than 5.0 mm in eight cases. With conventional angiography used as the reference standard, CT angiography has one false-positive and two false-negative findings, resulting in a sensitivity of 88% (15/17) and a specificity of 89% (8/9). For 12 of 15 aneurysms, CT angiography was rated equal or superior to conventional angiography in depicting all aspects (shape, orientation, neck, and parent vessel) of the aneurysms. CONCLUSION: We were able to detect aneurysms of the circle of Willis as small as 2 mm in size. Using conventional angiography as the reference standard, CT angiography has a sensitivity of 88% and a specificity of 89% for the detection of aneurysms in the circle of Willis. In almost all cases, CT angiography was equal or superior to conventional angiography in characterizing the aneurysms for surgical planning. PMID- 7484597 TI - Intracranial metastatic melanoma: correlation between MR imaging characteristics and melanin content. AB - OBJECTIVE: Preliminary reports based on limited numbers of cases have proposed that specific MR imaging patterns may permit a distinction between melanotic and amelanotic brain metastases in melanoma patients. The purpose of this study was to test this hypothesis by categorizing MR images obtained from a large series of patients and correlating the results with the percentage of melanin-containing cells in surgically resected metastases. SUBJECTS AND METHODS: The MR images of 30 patients with histologically proven intracerebral melanoma were reviewed retrospectively. Precontrast MR images were obtained with T1-weighted spin-echo sequences in axial and sagittal sections and with proton density-weighted and T2 weighted sequences in axial sections. After IV injection of gadopentetate dimeglumine (0.1 mmol/kg of body weight), T1-weighted images were obtained in axial and coronal sections. All patients had undergone gross total resection of the evaluated lesions. MR images of the metastases were reviewed and sorted into four groups on the basis of putative patterns: (1) melanotic pattern- hyperintense in relation to cortex on T1-weighted images, hypointense in relation to cortex on T2-weighted images, and isointense or hyperintense in relation to cortex on proton density-weighted images; (2) amelanotic pattern--hypointense or isointense in relation to cortex on T1-weighted images and hyperintense or isointense in relation to cortex on T2-weighted and proton density-weighted images; (3) indeterminate, or mixed, pattern--MR imaging characteristics that did not conform to those of one of the first two categories; and (4) hematoma pattern -MR imaging features that exhibited only hematoma characteristics. Tissue sections from all evaluated lesions were independently reviewed by a neuropathologist (G.N.F.), and the percentage of melanin-containing tumor cells in each resected metastatic lesion was estimated. The MR imaging data and histologic data were then compared to assess the predictive value of the MR imaging patterns. RESULTS: Forty-two metastatic lesions were identified and categorized by MR imaging pattern as follows: 10 melanotic, 11 indeterminate (mixed), 16 amelanotic, and five hematoma. Correlation with histologic findings revealed that a majority (7/10) of lesions that exhibited a melanotic MR imaging pattern had more than 10% melanin-containing cells, over half (9/16) of lesions that exhibited an amelanotic MR imaging pattern contained histologically identifiable melanin (but always in less than 10% of cells), and lesions that exhibited a mixed MR imaging pattern were either amelanotic or contained less than 10% melanotic cells. Conversely, a majority of lesions containing more than 10% melanotic cells (7/8) demonstrated the typical melanotic MR imaging pattern, lesions with less than 10% melanin-containing cells exhibited a variety of MR imaging patterns, and only about half of patients with amelanotic lesions (6/13) showed the characteristic amelanotic MR imaging pattern. For five lesions, potentially informative imaging data on melanin content was obscured by histologically documented hematoma formation. CONCLUSION: Only a minority of melanoma metastases have the anticipated MR imaging findings of melanotic melanoma, which consist of high signal intensity relative to that of cortex on T1 weighted images and low signal intensity relative to that of cortex on T2 weighted images. Of tumors that do exhibit this melanotic pattern, the majority have more than 10% melanin-containing cells. The putative MR imaging pattern for amelanotic melanoma is nonspecific, as over half of tumors with this pattern contain melanin. PMID- 7484598 TI - Iatrogenically induced intracranial hypotension syndrome. PMID- 7484600 TI - 95th anniversary of the ARRS: history of the Journal editors. American Roentgen Ray Society. AB - The change of AJR editorship with next month's issue and the celebration of the centennial year of the X-ray make this an appropriate time to acknowledge the many distinguished editors who have served the American Roentgen Ray Society. A review of these gentlemen's service to this Journal is in no small measure a chronology of the Society's history itself, especially during its first turbulent decade. The recounting of this story should entertain and educate as well as pay tribute to these fine scholars. Perhaps most of all, it should cause us all to reflect upon the tremendous sacrifices made for radiology in the United States and for our Society by these individuals. PMID- 7484599 TI - Transluminal balloon dilatation of the lacrimal drainage system for the treatment of epiphora. AB - OBJECTIVE: The purpose of this study was to evaluate the efficacy of transluminal balloon dilatation done under digital subtraction fluoroscopic monitoring in the treatment of incomplete and complete obstruction of the lacrimal drainage system (LDS) in patients with epiphora. SUBJECTS AND METHODS: Transluminal balloon dilatation of the LDS was attempted in 19 eyes with incomplete and 61 eyes with complete obstruction, including six cases of anastomotic obstruction after failed dacryocystorhinostomy. A soft plastic arterial sheath over a stump-guiding metal probe was introduced through the superior canaliculus into the nasolacrimal apparatus. A 0.016-in. specially angled steerable guidewire was introduced through the sheath and across the site of obstruction, advanced into the nasal cavity, and brought forward through the external nare. A low-profile angioplasty balloon dilatation catheter was passed retrograde over the guidewire through the nasal aperture to the site of obstruction and inflated for dilatation. RESULTS: Technical success was achieved in all 19 eyes with incomplete obstruction and in 46 (75%) of 61 eyes with complete obstruction. Initial success was achieved in all 19 eyes with incomplete obstruction and in 36 (59%) of 61 eyes with complete obstruction. In a follow-up period of 6-18 months (mean, 13 months), reobstruction occurred in none of the eyes with incomplete and in two eyes with complete obstruction. CONCLUSION: Our experience shows that transluminal balloon dilatation is an effective treatment for incomplete obstruction of the LDS. The procedure has a high failure and recurrence rate when the obstruction is complete. PMID- 7484601 TI - The life of Wilhelm Conrad Roentgen. AB - This year, 1995, marks the centennial of the discovery of X-rays by Wilhelm Conrad Roentgen on the historical date of that discovery, November 8, 1895 [1]. After completing my radiology residency at Brooke General Hospital, I was assigned to the U.S. Army 5th General Hospital in Stuttgart Bad Cannstatt, West Germany. This period of payback was from October 1966 to August 1969. During these memorable years, the opportunity presented itself for me to make a pilgrimage to every location associated with the life of Roentgen. It is my pleasure to share that pilgrimage with you. PMID- 7484603 TI - What is the risk of reaction on repeat exposure to contrast material, and how should the patients be premedicated? PMID- 7484602 TI - American martyrs to radiology. John Townsend Pitkin (1858-1935). 1936. PMID- 7484604 TI - Contrast-enhanced CT examination on a patient who has previously developed hives. PMID- 7484605 TI - Unenhanced CT scans in identifying hematomas after blunt abdominal trauma. PMID- 7484606 TI - Is it necessary to routinely use gadolinium in the MR imaging evaluation of soft tissue tumors of the extremities? PMID- 7484607 TI - When is it appropriate to do enhanced MR imaging of the brain? PMID- 7484608 TI - Danger in performing MR imaging on women who have tattooed eyeliner or similar types of permanent cosmetic injections. PMID- 7484609 TI - Interventional radiology of the chest. PMID- 7484610 TI - Short rib(s) PMID- 7484612 TI - Mammographic follow-up of clustered microcalcifications. PMID- 7484611 TI - Mammography after conservative treatment of breast cancer. PMID- 7484613 TI - Pancreaticopleural fistula diagnosed sonographically. PMID- 7484614 TI - Appendiceal mucocele: sonographic and MR imaging findings. PMID- 7484615 TI - An unusual manifestation of xanthogranulomatous pyelonephritis: bilateral focal solid renal masses. PMID- 7484617 TI - Placental polyp: MR imaging findings. PMID- 7484616 TI - Renal artery stenosis with contralateral renal cell carcinoma: an indication for prophylactic angioplasty? PMID- 7484618 TI - Rotator cuff disease. PMID- 7484619 TI - Intraosseous neurilemmoma of the lower third of the fibula. PMID- 7484620 TI - The value of CT in determining potential instability of simple wedge-compression fractures of the lumbar spine. AB - PURPOSE: To determine whether plain films alone are sufficient in the evaluation of stability of simple wedge-compression fractures of the lumbar spine. METHODS: Plain films and CT scans of 53 consecutive patients seen during a 2-year period with lumbar spine fractures were retrospectively reviewed. Six readers blinded to the CT diagnosis independently read each patient's plain films. Plain-film findings were scored on a five-point graded response scale using criteria proposed by Gehweiler and Daffner. In addition, a fracture was considered to be possibly unstable if there was involvement of more than one vertebral level or greater than 50% loss of anterior vertebral body height. CT findings represented the standard for comparison. CT scans were independently evaluated by three additional readers. Two-column involvement, middle-column involvement alone but with retropulsion, multiple-level involvement, or greater than 50% loss of vertebral height indicated potential instability. RESULTS: For 14 stable and 39 potentially unstable lumbar spine fractures, the pooled (mean) plain-film negative predictive value for detection of potentially unstable fractures was 0.62 (95% confidence interval, 0.53 to 0.70), with a sensitivity of 0.83 (95%, confidence interval; 0.78 to 0.87), and specificity of 0.80 (95% confidence interval, 0.70 to 0.87). CONCLUSION: Plain films are not adequate for determining stability of lumbar spine fractures. PMID- 7484621 TI - Lumbar spondylolysis without spondylolisthesis: recognition of isolated posterior element subluxation on sagittal MR. AB - PURPOSE: To document the occurrence of isolated dorsal subluxation of posterior elements in cases of lumbar spondylolysis without spondylolisthesis both quantitatively (using spinal canal measurements) and qualitatively (by visual inspection) on sagittal MR images. METHODS: Retrospective analysis identified 63 patients with lumbar spondylolysis (confirmed by CT or conventional radiography) who had undergone MR imaging. From these we identified 12 patients with pars interarticularis defects but no evidence of spondylolisthesis. Measurements of anteroposterior spinal canal diameters were performed in these 12 patients to ascertain whether the sagittal canal diameter at the level of the spondylolysis exceeded the normal range as determined from 100 control subjects. RESULTS: In 9 of 12 patients the spinal canal was abnormally widened at the level of the spondylolysis because of dorsal subluxation of posterior elements. In 5 of these patients, the subluxation was readily visible on midline sagittal MR images. In 4 patients, spinal canal measurements were necessary to document this phenomenon. CONCLUSION: In the majority of patients with spondylolysis but without spondylolisthesis, sagittal MR images can show isolated dorsal subluxation of posterior spinal elements. PMID- 7484622 TI - Sciatic endometriosis: MR evaluation. PMID- 7484623 TI - Cervical myelopathy secondary to Hunter syndrome in an adult. AB - We present a case of type II mucopolysaccharidosis in which the diagnosis was delayed until the onset of cervical myelopathy in adulthood. Radiographic features were characteristic, with striking dural thickening shown on CT and MR imaging. PMID- 7484624 TI - Juvenile Huntington disease: CT and MR features. AB - PURPOSE: To describe the clinical and radiologic manifestations of juvenile Huntington disease and to determine whether adult imaging criteria for Huntington disease are helpful for pediatric patients. METHODS: Six patients (3 to 18 years of age; mean age, 9.8 +/- 5.6 years; 3 female, 3 male) with juvenile Huntington disease were studied with CT (n = 6) and/or MR (n = 3). CT and MR studies were evaluated for frontal horn distance/intercaudate distance and bicaudate ratios, which were compared with those of 24 age-matched healthy children and 12 age matched patients with Leigh (n = 9) or Wilson (n = 3) disease. RESULTS: Atrophy of the caudate nuclei was identified in all Huntington patients. The frontal horn distance/intercaudate distance (1.64 +/- 0.39) and bicaudate (0.205 +/- 0.060) ratios of the patients with juvenile Huntington disease were found to be significantly different from those of healthy children and that of those patients with Leigh/Wilson disease. The 3 patients with Huntington disease who underwent MR evaluation were noted to have increased proton density- and T2-weighted signal in the caudate nuclei and putamina. CONCLUSION: As in adult patients, the use of frontal horn distance/intercaudate distance and bicaudate ratios are helpful for the diagnosis of Huntington disease in pediatric patients. On MR, increased proton density- and T2-weighted signal in the atrophic caudate nuclei and putamina are additional features of juvenile Huntington disease. PMID- 7484625 TI - Normal MR appearance of the pituitary gland in the first 2 years of life. AB - PURPOSE: To investigate changes in the size, shape, and signal intensity of the pituitary gland during the first 2 years of life. METHODS: One hundred consecutive MR studies, spin-echo T1- and T2-weighted sequences, of children 0 to 2 years of age were analyzed. Signal intensity and shape of the gland were graded and its signal intensity measured. RESULTS: There was a statistically significant difference in the signal intensity of both the anterior and posterior pituitary gland and in the shape between children younger than and older than 6 weeks of age and younger than and older than 27 weeks of age. There were definite trends in the size of the gland with age (decrease in height and increase in anteroposterior diameter and width). CONCLUSIONS: There are changes in the pituitary gland in the newborn period that may reflect the ongoing changes in pituitary hormones reported to occur during this time. PMID- 7484626 TI - Ventriculus terminalis of the conus medullaris: MR findings in children. AB - PURPOSE: To determine the appearance of the ventriculus terminalis of the conus medullaris and the frequency with which it is seen on MR. METHODS: Four hundred eighteen normal spine MR examinations were reviewed. RESULTS: Eleven (2.6%) of 418 children, all younger than 5 years of age, demonstrated an ovoid, nonenhancing, smooth dilation of the central canal within the conus medullaris. Averaged volume of the oblate spheroid was 0.18 cm3. Fluid within the cavity behaved as cerebrospinal fluid on all pulse sequences, and there was no abnormal signal in the cord tissue surrounding the cavity. CONCLUSION: Asymptomatic localized dilatation of the ventriculus terminalis is a normal developmental phenomenon that can be seen on MR. PMID- 7484627 TI - Infantile-onset spinocerebellar ataxia: MR and CT findings. AB - PURPOSE: To report the MR and CT findings in a hereditary disease, infantile onset spinocerebellar ataxia (IOSCA). METHODS: We studied the brains of 17 patients with infantile-onset spinocerebellar ataxia with CT and/or MR to determine the presence of cerebellar and brain stem atrophy and parenchymal lesions. RESULTS: Cerebellar cortical atrophy was seen in 13 patients. The degree of atrophy correlated with increasing age and clinical deterioration. Brain stem atrophy was seen in 8 patients. It was never severe, and the basis pontis was not flattened even in the most severe cases. Hyperintense lesions were noted within the white matter of cerebellum, in the dentate nuclei, and in the middle cerebellar peduncles in 3 patients. The upper cervical cord was seen in 9 patients and showed mild to moderate atrophy in 4. The basal ganglia and cerebral hemispheres were normal, except in 2 patients transient cortical and subcortical lesions developed during episodes of status epilepticus; mild cortical brain atrophy subsequently developed. CONCLUSION: The brain MR and CT findings of patients with infantile-onset spinocerebellar ataxia correspond to the neuropathologic entities of cerebellar cortical atrophy, olivopontocerebellar atrophy, and spinocerebellar atrophy. The appearance of the findings followed a uniform time sequence from cerebellar cortical atrophy in the early stage of the disease to olivopontocerebellar atrophy and spinocerebellar atrophy in the later stage. The severity of atrophy correlated with clinical deterioration. PMID- 7484628 TI - Detection of discrete white matter lesions after irreversible compression of MR images. AB - PURPOSE: To validate the use of techniques of irreversible compression of images, which can be performed using a block-based discrete cosine transform technique as defined by the Joint Photographic Experts Group, before they can be used in clinical applications, by evaluating the effect of compression on the ability of observers to detect discrete white matter lesions on MR images of the brain. METHODS: Sixty T2- and intermediate-weighted spin-echo images were compressed with varying degrees of coefficient quantization with compression ratios from 1:1 to more than 40:1, randomized, and evaluated by three observers blinded to the degree of compression. RESULTS: No significant difference in the number of lesions detected was apparent until compression ratios reached 40:1, despite a significant subjective loss in perceived image quality at 20:1. Only small (< or = 5 mm) lesions were missed at the highest degree of compression. No significant differences were observed in the detection of confluent periventricular white matter disease at any degree of compression tested. CONCLUSIONS: The use of high degrees of irreversible compression of MR images may be acceptable for diagnostic tasks. PMID- 7484629 TI - MR volume estimation of subcortical brain lesions and ventricular cerebrospinal fluid: a simple and accurate stereologic method. AB - PURPOSE: To describe an MR imaging quantification method for estimation of total volumes of both white and gray matter subcortical lesions and ventricular cerebrospinal fluid (CSF) in the living human brain, and to determine the method's reliability. METHODS: In 12 subjects, total subcortical lesion and ventricular CSF volumes were estimated using systematic sampling. Systematic sampling was performed on equidistant MR sections using a counting grid with systematically ordered intersection points. The grid was randomly positioned on each consecutive MR section. Each grid intersection point hitting the structure of interest represents a fixed known volume dependent on grid intersection point distance and the sum of the section thickness and section gap. RESULTS: Total volume estimation of subcortical lesion and ventricular CSF takes 15 and 5 minutes per subject, respectively. Coefficients of error of the individual volume estimates ranged from .01 to .13 and are negligible to the coefficients of the group mean (range, .70 to .89). For subcortical lesion volume, the random intraobserver error yielded .04 and for ventricular CSF .02; the random interobserver error amounted to .11 and .04, respectively; and the systematic interobserver error was .15 and .04, respectively. CONCLUSION: The method described here for subcortical lesion and ventricular CSF volume estimation is accurate, reliable, valid, and fast. PMID- 7484630 TI - Brain slice holder for MR. AB - We made an MR-compatible brain slice holder that ensures that the images have the same location and orientation as the subsequent histologic sections. This device encloses the unfixed tissues, thus reducing the danger of exposure to pathogens. MR imaging time is saved by scanning selected slices rather than the whole brain; this technique is especially useful when brain sections at varying angles, rather than parallel slices, are taken. PMID- 7484631 TI - Sizing rings: a simple technique for measuring intracranial lesions. PMID- 7484632 TI - Balloon test occlusion of the internal carotid artery with hypotensive challenge. AB - PURPOSE: To evaluate the usefulness of provocative testing with hypotensive challenge during balloon test occlusion of the internal carotid artery before carotid sacrifice and to correlate tolerance of balloon test occlusion with clinical outcome after carotid artery sacrifice. METHODS: Forty-seven consecutive cases of balloon test occlusions performed at our institution during the past 4 years were retrospectively reviewed. Occlusion was performed under normotensive conditions with distal perfusion of heparinized saline for 20 minutes, or until a deficit was perceived. If 20 minutes of normotension was tolerated, hypotension was induced to two thirds of mean arterial pressure for 20 minutes, or until a deficit was perceived. RESULTS: Of 47 patients, 4 (9%) had deficits at normotension. Of the remaining 43 patients, 9 (21%) had deficits at hypotension. One patient with a positive hypotensive test occlusion underwent carotid artery sacrifice after extracranial-intracranial bypass without sequelae. In one of the 19 patients who clinically tolerated test occlusion with hypotension and had carotid sacrifice (surgical ligation of the intracranial carotid artery), a mild embolic stroke developed, probably from the giant carotid wall aneurysm. This patient fully recovered; MR imaging showed mild changes consistent with emboli distal to the aneurysm. Symptomatic complications were noted in 2 (4%) patients, and asymptomatic arterial dissections were noted in 3 (6%) patients. CONCLUSION: Balloon test occlusion with hypotensive challenge is safe, economical, and greatly increases the sensitivity of balloon test occlusion. The predictive value of a negative test is high. However, to determine the test's specificity compared with quantitative imaging, controlled trials will be necessary. PMID- 7484633 TI - Immediately detachable coil for aneurysm treatment. AB - We developed an endovascular coil that is instantly detached by high-frequency electrical current. By applying the electrical current, a polyvinyl alcohol junction between the coil and the delivery wire is disrupted by heat within a second. This detachment system was shown to be reliable in the treatment of experimental aneurysms. PMID- 7484635 TI - The 'gooseneck' concept in microvascular retrieval. PMID- 7484634 TI - Endovascular therapy of intractable epistaxis complicated by carotid artery occlusive disease. AB - Three cases of intractable spontaneous posterior epistaxis refractory to nasal packing and complicated by ipsilateral carotid artery occlusive disease were successfully treated with internal maxillary artery occlusion with microcoils. There were no complications and no recurrent episodes of epistaxis at a mean follow-up of 12 months. The presence of ipsilateral carotid artery disease requires modification of standard distal internal maxillary artery embolization because of the recruitment of external carotid to internal carotid and external carotid to ophthalmic artery collateral pathways, with subsequent risk of particle embolization of these arterial distributions. Proximal internal maxillary artery microcoil embolization eliminates this risk and is equivalent to surgical internal maxillary artery ligation. This procedure also provides additional information about the internal carotid artery collateral circulation. PMID- 7484636 TI - Characterization of multiple sclerosis plaques with T1-weighted MR and quantitative magnetization transfer. AB - PURPOSE: To investigate the relationship between the appearance of multiple sclerosis lesions identified on unenhanced T1-weighted images and their corresponding magnetization transfer ratios. METHODS: A total of 119 white matter lesions seen on T2-weighted images in 17 patients with multiple sclerosis were evaluated. Axial T1-weighted images were used to classify the lesions as isointense to white matter (10 lesions), hypointense to white matter but hyperintense to gray matter (44 lesions), hypointense to gray matter (59 lesions), and relatively isointense to cerebrospinal fluid (6 lesions). The magnetization transfer ratio of each lesion was calculated, and an average magnetization transfer ratio for each subcategory was determined. RESULTS: The magnetization transfer ratio values became progressively lower with increasing hypointensity of lesions on T1-weighted images. The average magnetization transfer ratio for lesions isointense to white matter, hypointense to white matter but hyperintense to gray matter, hypointense to gray matter, and relatively isointense to cerebrospinal fluid was 34.90 +/- 2.67 mean +/- SD), 30.93 +/- 3.57, 27.27 +/- 3.56, and 23.62 +/- 2.83, respectively. All groups were significantly different from each other. CONCLUSION: Lesions isointense to white matter exhibited higher magnetization transfer ratio values than lesions that were hypointense. These findings are consistent with relative preservation of the myelin structure in the former, perhaps indicating that these lesions are predominantly inflammatory (edematous) in nature. The proportionately lower magnetization transfer ratio values of lesions that appear progressively more hypointense on T1-weighted images may reflect varying degrees of demyelination, with increasing lesion hypointensity corresponding to more breakdown in the macromolecular structure. These results suggest that T1-weighted images may be useful in characterizing the underlying pathologic substrate in multiple sclerosis plaques. PMID- 7484637 TI - The evolution of multiple sclerosis lesions on serial MR. AB - PURPOSE: To characterize temporal changes in signal intensity patterns of multiple sclerosis lesions on serial MR. METHODS: T1-, T2-, proton density-, and contrast-enhanced T1-weighted MR was performed on five patients with relapsing remitting multiple sclerosis at least 22 times in the course of 1 year. RESULTS: Forty-three enhancing lesions and 1 new lesion that never showed enhancement were detected and followed for periods ranging from approximately 4 weeks to 1 year (total of 702 time points). At first detection the center of new lesions was brighter than the periphery (20 of 24 new lesions on proton density-weighted and 19 of 23 new lesions on contrast-enhanced images). On contrast-enhanced images, ring hyperintensity was predominant at time points later than 29 days. As lesions aged, a residual rim of "nonenhancing" hyperintensity often was noted on contrast enhanced images. Some older lesions (> 1 year) showed similar appearance on unenhanced T1-weighted images. On proton density-weighted images ring hyperintensity was most frequent 2 to 4 months after lesion detection. The estimated average duration of gadopentetate dimeglumine enhancement was 1 to 2 months. CONCLUSIONS: A lesion evolution pattern relevant to MR was inferred. We believe that specific information about the histopathologic evolution of a lesion may be extracted not only from contrast-enhanced but also from nonenhanced serial MR. Assessment of drugs targeting specific phases of lesion evolution could benefit from quantitative pattern analysis of routine MR images. PMID- 7484638 TI - Methanol intoxication with putaminal and white matter necrosis: MR and CT findings. AB - We report a case of methanol intoxication in which the initial CT scans appeared normal. MR at 4 days showed the typical putaminal lesions of methanol intoxication and, in addition, peripheral white matter lesions that spared a thin rim of subcortical white matter. A CT scan at 17 days showed the putaminal and white matter lesions. The white matter lesions correspond well to lesions previously described in pathologic specimens. PMID- 7484639 TI - Subcortical white matter lesions in osmotic demyelination syndrome. AB - We describe a case of histologically proved osmotic demyelination syndrome. In addition to abnormal T2 signal within the pons and thalami, MR showed linear enhancing lesions at the cortical medullary junction. PMID- 7484640 TI - Bilateral occipital calcification associated with celiac disease, folate deficiency, and epilepsy. AB - In a patient with celiac disease, folate and iron deficiency, and epilepsy, CT over a 4-month period showed parietoocipital calcifications in the corticomedullary junctions. The calcification was progressive, but it and the seizures stabilized after institution of a gluten-free diet and iron and folic acid supplements. PMID- 7484641 TI - Meningeal myelomatosis: CT and MR appearances. AB - Meningeal myelomatosis is a rare feature of multiple myeloma. We report a case of IgG-kappa myeloma presenting as bilateral intracranial extraaxial masses. PMID- 7484642 TI - Intratentorial lipomas with Meckel's cave and cerebellopontine angle extension. AB - An unusual case of bilateral intratentorial lipomas with extension into Meckel's caves and the cerebellopontine angle is described. Surgical and histopathologic correlation demonstrate that the lipoma encased the trigeminal nerve in Meckel's caves. The origin of the lipoma from the anteromedial margins of the tentorium is discussed and correlated with a recently proposed theory for the development of intracranial lipomas. PMID- 7484644 TI - Primary rhabdomyosarcoma of the scalp. PMID- 7484645 TI - The dural tail sign revisited. PMID- 7484646 TI - Postoperative imaging of the multichannel cochlear implant. AB - PURPOSE: To present the postoperative radiographic appearance of the multichannel cochlear implant and to suggest criteria for the radiographic evaluation of postimplant patients. METHODS: One hundred forty-one patients, 73 female and 68 male, had multichannel cochlear implants inserted. One hundred thirty-five intraoperative radiographs, 31 postoperative radiographic examinations, and 10 postoperative CT examinations were retrospectively reviewed. RESULTS: In 135 patients, a Stenver's or anteroposterior projection confirmed the normal placement of the implant by demonstrating the electrodes to be regularly spaced and gently curved within the first turn of the cochlea. The insertion depth was determined by counting the number of electrodes that projected medial to the cochlear promontory. In 5 patients in whom intracochlear placement of the electrode array could not be confirmed on plain radiographs, CT demonstrated the location of the electrode array. In 3 patients with postoperative infections, CT either correctly identified or excluded the presence of a collection beneath the implant. CONCLUSION: Plain radiographs of the temporal bone are sufficient for the postoperative treatment of the majority of postimplant patients. CT should be performed when plain radiographs cannot adequately show the location of the electrode array or if postoperative infection is suspected. PMID- 7484647 TI - Reformatted planar 'Christmas tree' MR appearance of the endolymphatic sac. AB - A high-resolution three-dimensional Fourier transform technique and prototype bilateral dual phased-array surface coil technique was used to make inner ear structures visible on MR. Multiplanar reformatted images, parallel to the plane of the vestibular aqueduct, allowed viewing of the entire endolymphatic sac/vestibular aqueduct on one section, producing a "Christmas tree" shape. The reformation was obtained using a double oblique angle, 45 degrees from true sagittal and 70 degrees from the orbital-meatal axis. PMID- 7484643 TI - MR and positron emission tomography with fludeoxyglucose F 18 in gliomatosis cerebri. AB - A 16-year-old girl presented with a unilateral third nerve palsy and predominant gray matter involvement on MR and positron emission tomography with fludeoxyglucose F 18. These findings were manifestations of gliomatosis cerebri. The antemortem diagnosis was made by partial temporal lobectomy. PMID- 7484650 TI - Multifocal intracranial occlusive vasculopathy resulting in stroke: an unusual manifestation of Williams syndrome. PMID- 7484648 TI - Recurrent parotid adenocarcinoma occurring within the cerebellopontine angle. AB - A patient presented 3 1/2 years after resection of a malignant parotid tumor with a new cerebellopontine angle mass. The presence of multiple punctate calcifications and loss of T2 signal were helpful in correctly identifying the lesion as mucinous adenocarcinoma. PMID- 7484651 TI - Progressive venous occlusion in a neonate with Sturge-Weber syndrome: demonstration with MR venography. AB - Progressive cerebral sinovenous occlusion in a neonate with Sturge-Weber syndrome was documented by using two-dimensional time-of-flight MR venography. There was no evidence of intraluminal thrombus on routine spin-echo images obtained either before or after the onset of seizures, despite MR venographic evidence in both studies of venous abnormalities. PMID- 7484649 TI - Spurious hyperfixation of hexamethylpropyleneamine oxime in acute embolic stroke. AB - A 71-year-old man with atrial fibrillation suddenly had an embolism in the right middle cerebral artery. Single-photon emission CT was performed 4 hours after the onset of stroke and revealed markedly elevated 99mTc-hexamethylpropyleneamine oxime (HMPAO) uptake in the right frontotemporoparietal region. This area of HMPAO hyperfixation corresponded to the area of infarction, as determined by CT and MR. Our findings showed that HMPAO single-photon emission CT overestimated reflow hyperemia after reperfusion in the acute stage of embolic stroke, as it does in the subacute stage. PMID- 7484652 TI - The association between tuberous sclerosis and insulinoma. AB - We describe a patient with tuberous sclerosis and an insulinoma. The neurologic abnormalities typically present in patients with tuberous sclerosis may, in rare cases, be manifestations of hypoglycemia. We discussed a possible association between tuberous sclerosis and multiple endocrine neoplasia type I. PMID- 7484653 TI - Duplication of the extracranial internal carotid artery. AB - We describe a case of duplication of the left internal carotid artery from a point 1 cm distal to the origin to the proximal petrous segment where the vessel reunites. Duplication and fenestration of the internal carotid artery are discussed. A review of embryologic development is presented. Identification of these entities is important, especially in patients who require surgical intervention involving the internal carotid artery. PMID- 7484654 TI - Dolichoectasia of the anterior cerebral arteries in an adolescent. AB - We report a case of bilateral dolichoectasia of the anterior cerebral arteries in a 14-year-old girl as documented by MR imaging and cerebral angiography. Concurrent thickening of the adjacent medial frontal lobes was found, suggesting either a neuronal migrational abnormality or secondary changes attributable to vascular insufficiency. PMID- 7484655 TI - Frontoethmoidal giant cell reparative granuloma. AB - We present a case of a giant cell reparative granuloma of the frontoethmoidal region that had a large intracranial extraaxial component and was studied with MR. Although rare, giant cell reparative granuloma can be suggested in the correct clinical setting and when MR features suggest a fibrous lesion. PMID- 7484656 TI - Spinal cord multiple sclerosis and devic neuromyelitis optica in children. PMID- 7484657 TI - Are sagittal T2-weighted fast spin-echo MR images sufficient for routine spine imaging? PMID- 7484658 TI - The potential of magnetization transfer MR to differentiate fibrillary gliosis from fibrous gliosis. PMID- 7484659 TI - Radiation dose to the lens from cerebral arteriography. PMID- 7484661 TI - Midbrain lesions and third-nerve palsy. PMID- 7484660 TI - Differentiating cerebral lymphoma from nonmalignant central nervous system lesions in patients with AIDS. PMID- 7484662 TI - Retained sponge in the neck: MR appearance. PMID- 7484663 TI - Annotated bibliography. PMID- 7484664 TI - Drug use and crime in Mobile, Alabama, 1991-1992. PMID- 7484666 TI - The Retinal Research Team at the Eye Foundation Hospital/UAB. AB - The trustees of the Eye Foundation, Inc. and the Eye Foundation Hospital constitute an independent institution, which is associated with the University of Alabama at Birmingham only in research and resident training. Clarence Blair is the Chairman of the Foundation Board. Harold Skalka, M.D., F.A.C.S., is the Chairman of the Department of Ophthalmology, and his offices are in the Eye Foundation Hospital building. With funds raised largely by Alston Callahan, M.D., F.A.C.S., Director of Development, and with the assistance of fellow trustees, the retina research department has been greatly expanded in the last few years. In this article, the seven prestigious retinal investigators are introduced and short summaries of their research aims are presented. All work toward a better understanding of clinical problems which baffle ophthalmologists, as macular degeneration and diabetic eye disease. PMID- 7484667 TI - There is some Luddite in all of us. PMID- 7484665 TI - The diaries of Dr. Samuel Patton Hand 1911-1917. Part I. AB - Dr. Samuel Patton Hand practiced medicine in Demopolis, Alabama, for many years before his death in 1917. During the last seven years of his life he kept detailed diaries. These diaries have been passed down through the family since his death. These papers review the diaries, which present an intimate look at the life of an Alabama small-town doctor during this period. This first paper deals with the doctor's daily personal and social life, and looks at the transportation in Dr. Hand's day and its effect on his medical practice. The next paper will examine purely medical matters. A third paper will look at excerpts from a notebook or "Pearl book" the doctor kept. PMID- 7484668 TI - The Alabama Medicaid DUR Program. PMID- 7484670 TI - Medical research, past, present, and future. PMID- 7484669 TI - Physicians as agents. PMID- 7484671 TI - VBAC clinical policy: an AAFP guideline. PMID- 7484672 TI - Thoughts on the varicella vaccine. PMID- 7484673 TI - Preventing chronic renal failure in diabetes. PMID- 7484675 TI - Use of a metal detector to track swallowed metal objects. PMID- 7484674 TI - TOXNET: the National Library of Medicine's toxicology database. PMID- 7484676 TI - St. Anthony's fire revisited. PMID- 7484677 TI - St. Anthony's fire revisited. PMID- 7484679 TI - Interstitial cystitis misdiagnosed as benign prostatic hyperplasia. PMID- 7484678 TI - Ebola virus and HIV: a contrast in public health measures. PMID- 7484680 TI - Basal cell carcinoma of the leg. PMID- 7484681 TI - Patterns of psychoactive substance use among adolescents. PMID- 7484682 TI - Helicobacter pylori and gastrointestinal disease. AB - Helicobacter pylori infection is quite common. In the United States, prevalence varies considerably with race, nationality, socio-economic status and location of residence. In Western countries, the prevalence of the infection has shown a steady increase with increasing age. H. pylori has been shown to cause chronic gastritis. Most patients infected with H. pylori are asymptomatic and require no therapy. The precise role of the infection in the pathogenesis of peptic ulcer disease is unknown. However, H. pylori infection is associated with a high recurrence rate of both gastric and duodenal ulcers. Eradication of the infection reduces the recurrence rate. Once H. pylori infection is acquired, it usually persists for years, possibly for the patient's lifetime. Although a causative role in gastric cancer has not been proved, evidence suggests an association between H. pylori infection and well-differentiated gastric adenocarcinoma and gastric lymphoma. PMID- 7484683 TI - Pseudotumor of infancy and congenital muscular torticollis. AB - Pseudotumor of infancy presents as a discrete, firm mass in the distal sternocleidomastoid muscle in infants two to four weeks of age. Congenital muscular torticollis may develop from the resultant fibrosis in 10 to 20 percent of cases. Hip dysplasia is an associated feature of congenital muscular torticollis in approximately 10 percent of cases. Pseudotumor of infancy must be differentiated from other causes of cervical soft tissue masses. Diagnostic choices include fine-needle aspiration biopsy, cervical radiography, ultrasonography, computed tomographic scanning of the head and neck, and magnetic resonance imaging. Left untreated, congenital muscular torticollis may lead to significant craniofacial asymmetry and scoliosis. Heat, massage and passive stretching exercises are the preferred initial treatments for pseudotumor and torticollis. More than 70 percent of patients will respond to this approach. Surgery should be reserved for treatment of cases that persist past the first year of life. PMID- 7484684 TI - Resistant respiratory pathogens and extended-spectrum antibiotics. AB - Traditional antibiotics such as amoxicillin, tetracycline and erythromycin remain the drugs of first choice for most bacterial respiratory infections. However, the usefulness of these agents varies, depending on local bacterial resistance patterns and patient factors. In the United States, amoxicillin and penicillin resistance currently occurs in 20 to 30 percent of Streptococcus pneumoniae strains, 30 to 40 percent of Haemophilus influenzae strains and 70 to 90 percent of Moraxella catarrhalis strains. For infections with these pathogens, selective use of the newer extended-spectrum oral antibiotics may be indicated. Cefuroxime axetil (a second-generation cephalosporin), cefpodoxime (a third-generation cephalosporin), amoxicillin-clavulanate (a beta-lactamase inhibitor combination agent) and clarithromycin or azithromycin (extended-spectrum macrolides) are all relatively effective against organisms that are commonly resistant to penicillin and amoxicillin. PMID- 7484686 TI - Trial of labor vs. elective repeat cesarean section. AAFP Task Force on Clinical Policies for Patient Care. AB - A trial of labor after a previous low transverse cesarean section in the absence of ongoing contraindications is safe for most women. A comprehensive review and meta-analysis of the literature was conducted to determine outcomes, costs and women's preferences by method of delivery. About seven of 10 women who undergo a trial of labor after previous low transverse cesarean section can expect to deliver vaginally. The following differences in absolute risks were identified: a trial of labor was associated with a slightly increased risk of uterine rupture (0.24 percent) and a decreased risk of infection and fever (5.2 percent) and postpartum bleeding (0.59 percent) as compared with an elective repeat cesarean section. The difference in risk for newborns with an Apgar score of less than 7 at five minutes was 0.85 percent for infants delivered vaginally after a trial of labor versus infants delivered by elective repeat cesarean section. Financial cost (as measured by hospital charges) of cesarean delivery was 1.66 to 2.4 times greater than the cost of vaginal birth. This difference was due almost entirely to the longer length of hospital stay for women who had a cesarean delivery. When given the opportunity, about two-thirds of eligible women choose a trial of labor over elective repeat cesarean section. The reasons underlying women's preferences for a trial of labor or elective repeat cesarean section were diverse and changed during pregnancy. As a consequence, women's preferences for the method of delivery must be explored and respected throughout pregnancy and during the delivery process.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7484685 TI - Natural family planning. AB - A better understanding of reproductive anatomy and physiology has led to newer methods of natural family planning, including the ovulation method and the symptothermal method. Studies have shown that 1 percent of couples who consistently and correctly use either of these methods conceives during one year of use. However, in the absence of standardized interpretation of study results, use-effectiveness data of these methods vary widely. The lactational amenorrhea method may be used in selected patients during the first six months postpartum, and its effectiveness in avoiding pregnancy is reported to be as high as 98 percent. Advocates of natural family planning promote these methods as inexpensive and safe ways to determine periods of peak fertility and point to the potential benefit of increased communication in a relationship. PMID- 7484687 TI - When to suspect an acoustic neuroma. AB - Acoustic neuroma is a rare but important cause of sensorineural hearing loss. Patients with asymmetric hearing loss or unilateral tinnitus should be evaluated expeditiously to prevent further neurologic damage. Gadolinium-enhanced magnetic resonance imaging is the best tool for making the diagnosis of acoustic neuroma. Surgical removal using a translabyrinthine approach, is the favored treatment, although radiation therapy and expectant management may be chosen for patients at high surgical risk. PMID- 7484688 TI - Preventing disease progression in chronic renal failure. AB - Progression toward end-stage renal disease is usually inexorable in patients with diabetic and nondiabetic nephropathy. These patients can be identified at an early stage based on history, abnormal urinalysis or reduced glomerular filtration. Recent advances have made it possible to slow the progression of chronic renal failure. Major interventions include antihypertensive therapy, dietary protein restriction and, in patients with diabetes, strict glycemic control and angiotensin-converting enzyme inhibitor therapy. Collaboration with a nephrologist can help guide the family physician in the appropriate use of these modalities and help avoid common complications. Major efforts in slowing the progression of renal failure may lead to a decreased incidence of end-stage renal disease, with savings in morbidity, mortality and cost. PMID- 7484689 TI - Leg cramps: differential diagnosis and management. AB - Leg cramps are a common problem, especially in the elderly. The differential diagnosis is extensive and includes the following conditions: true cramps, such as those related to heat, hemodialysis and electrolyte disturbances, as well as idiopathic cramps (the most common type); contractures occurring in conditions such as metabolic myopathies and thyroid disease; tetany, which is usually related to electrolyte disturbances, and dystonias, such as occupational cramps and those related to antipsychotic medications. Other leg problems that are not cramps, such as restless legs syndrome and periodic leg movements, also must be distinguished. The etiology of idiopathic leg cramps is not clear. Treatments for leg cramps include stretching exercises, quinine sulfate and vitamin E, but no treatment is conclusively effective. Nonetheless, in many patients relief of symptoms is achieved with one or more of these treatments. PMID- 7484690 TI - ACE inhibitors: myocardial infarction and congestive heart failure. AB - Myocardial infarction and congestive heart failure are associated with high morbidity and mortality rates. The results of recent clinical trials support a beneficial role for angiotensin-converting enzyme (ACE) inhibitors in attenuating mortality from myocardial infarction and left ventricular dysfunction. Large scale survival studies provide practical evidence that the addition of an ACE inhibitor to conventional therapy with diuretics and digoxin can significantly reduce the number of hospital admissions and the risk of death related to progressive heart failure. PMID- 7484691 TI - Corticosteroid injection therapy for overuse injuries. AB - Local corticosteroid injection therapy may be considered for many conditions commonly treated by the primary care physician. Chronic overuse and flawed biomechanics may lead to inflammation in areas easily accessible to injected local corticosteroids, including bursae, tendon sheaths and joints. It is generally preferable to mix the steroid preparation with an anesthetic agent to minimize the injection pain, provide immediate symptom relief and confirm the diagnosis. Local complications of corticosteroid injection include tendon and ligament weakening, bacterial infections of joints and related structures, and subcutaneous tissue atrophy. Systemic complications are rare but reportedly include hyperglycemia and adrenal axis suppression. PMID- 7484692 TI - Fournier's gangrene: a urologic emergency. AB - Fournier's gangrene, a form of necrotizing fasciitis, is an uncommon, fulminant, rapidly progressing subcutaneous infection of the scrotum and penis, and may occur in all age groups. Most cases involve a mixed synergistic infection of aerobic and anaerobic bacteria, and occur as a result of one of three mechanisms: local trauma, extension from a perianal, periurethral or ischiorectal infection. Fournier's gangrene is a urologic emergency that requires prompt recognition and aggressive hemodynamic stabilization, treatment with parenteral broad-spectrum antibiotics and surgical debridement. This disease is associated with a high mortality rate; a high index of suspicion is essential for early recognition whenever a patient presents with scrotal discomfort and swelling. PMID- 7484693 TI - Orofacial manifestations of child abuse and neglect. AB - About one-half of all child abuse cases involve some form of orofacial injury. Common signs of physical abuse include fractures of the teeth or the maxilla, mandible and other facial bones, facial burns, lacerations of the lips and lingual frenum, and bite marks on the face and neck. Sexual abuse should be suspected if erythematous, ulcerative, vesiculopustular, pseudomembranous and condylomatous lesions are present on the lips, tongue, palate, face or pharynx. Dental neglect, a common form of child maltreatment, should be suspected if rampant caries and oral infection, bleeding and trauma persist despite the elimination of financial and transportation obstacles. Questioning the child and parents separately may help uncover an obvious discrepancy between the clinical findings and the history. Reporting suspected child abuse to the appropriate agency may protect the child from further injury and help the family obtain assistance to correct abusive practices and minimize the recurrence of abuse. PMID- 7484694 TI - A practical approach to hirsutism. AB - Women often express concern about what they consider to be excess body or facial hair. This surplus of hair may be normal or it may signal hypertrichosis or hirsutism. Hirsutism may be idiopathic, secondary to increased responsiveness of hair follicles to normal circulating levels of androgens, or it may result from an excess of androgens, which may be exogenous, or of ovarian or adrenal origin. The evaluation of hirsutism must include the identification, or exclusion, of androgen-producing tumors, but other extensive evaluation is controversial. Treatment includes local measures, antiandrogenic therapy and treatment focused on the underlying source of excess androgen. PMID- 7484695 TI - AAP recommends use of live attenuated varicella vaccine. PMID- 7484696 TI - Guidelines issued for prevention of opportunistic infections in persons with HIV.U.S. Public Health Service and Infectious Disease Society of America. PMID- 7484697 TI - Statement on fungal infection in HIV patients. PMID- 7484698 TI - Special service for education of physicians about HIV and AIDS. PMID- 7484699 TI - Panic disorder and the primary care physician. PMID- 7484700 TI - Preterm labor: ACOG guidelines for care. PMID- 7484701 TI - Diabetes and 'normal' lifestyle. PMID- 7484702 TI - Erythrasma and seborrheic dermatitis of the groin. PMID- 7484704 TI - Identifying substance abuse in primary care. AB - Family physicians have an opportunity to identify substance abusers years before they have medical complications or present for drug treatment. Occult substance abuse can be effectively identified by simple screening questions and careful attention to clinical indicators during the history and physical examination. Early intervention may have dramatic effects on substance-abusing behavior and may prevent the many adverse medical, psychologic and social effects of substance abuse in these patients. PMID- 7484703 TI - Fine-needle aspiration biopsy of the breast. AB - Fine-needle aspiration biopsy is a quick, accurate, cost-effective office procedure that increases the diagnostic usefulness of breast examination. It is a technique best used at initial discovery of a lesion. Its sensitivity, ranging from 90 to 97 percent in large series, depends on two factors: biopsy performance and interpretation. To attain a high sensitivity for this procedure, a team approach that includes the physician, pathologist and radiologist is essential. PMID- 7484705 TI - Common intestinal helminths. AB - Groups at risk for intestinal helminth infestation include international students and travelers, migrant laborers, refugees, children of foreign adoptions and the homeless. Symptoms are often minimal or vague, yet many such infections carry significant long-term morbidity. Enterobius vermicularis, the common pinworm, is usually manifested in children as anal itching and is best diagnosed by using the cellophane-tape test to recover eggs from the perianal skin. Treatment with mebendazole is indicated for the entire family. Trichuris, or whipworm, infection may be found with Ascaris infection, but whipworms are longer-lived and more likely to produce symptoms that mimic those of inflammatory bowel disease. Treatment for these parasites is mebendazole. Other significant helminthic infections, which are currently more common in immigrant populations, include hookworms (Necator americanus, Ancylostoma duodenale), threadworms (Strongyloides) and various tapeworms (Taenia, Diphyllobothrium and Hymenolepis species). PMID- 7484706 TI - Panic disorder. AB - Panic disorder is a chronic illness that affects at least 3 percent of the population. Panic disorder is associated with significant morbidity and an increased risk of suicide. Patients generally present with multiple somatic and psychologic complaints, including heart palpitations, chest pain, tremor, shortness of breath, choking, nausea or abdominal distress, dizziness, derealization, fear of losing control or going crazy, fear of dying, paresthesias, chills or hot flushes, headache, diarrhea, insomnia, chronic fatigue, anxiety and depression. To make the correct diagnosis, these symptoms must be evaluated carefully since they also occur with serious cardiovascular, pulmonary, endocrinologic and neurologic disorders. Many effective treatments are available, including tricyclic antidepressants, selective serotonin reuptake inhibitors, monoamine oxidase inhibitors, benzodiazepines such as alprazolam and clonazepam, and psychotherapy. PMID- 7484707 TI - Oral hypoglycemic agents in type II diabetes mellitus. AB - The patient with type II, or non-insulin-dependent, diabetes mellitus (NIDDM) is characterized by obesity and insulin resistance, with resultant hyperinsulinemia and hyperglycemia. Sulfonylureas are the chief therapy for patients with NIDDM; for a limited time, these agents stimulate increased insulin secretion. With chronic administration, sulfonylureas improve the diabetic patient's insulin activity by increasing cellular insulin receptors and reducing insulin postreceptor defects. Metformin, a drug in the biguanide class, is now approved for use in the United States. This drug does not stimulate insulin release but works by lowering glucose in peripheral tissues. It can be used alone or in combination with a sulfonylurea. With sulfonylureas and metformin, therapy for the patient with NIDDM can be more effectively tailored. PMID- 7484708 TI - ACOG releases technical bulletin on preterm labor. PMID- 7484710 TI - Prenatal diagnosis: more than meets the eye. PMID- 7484709 TI - Glaucoma screening: a golden opportunity. PMID- 7484711 TI - Corporate takeover of medicine. PMID- 7484712 TI - Over-the-counter cimetidine and theophylline interaction. PMID- 7484713 TI - Delayed recurrence of thrombotic thrombocytopenic purpura. PMID- 7484714 TI - Pathogens in subdural empyema. PMID- 7484715 TI - Cholesterol and lipid screening in the fasting state. PMID- 7484716 TI - Asthma: diagnosis and management. AB - The prevalence of asthma is increasing in the United States, with an estimated 9 to 12 million persons affected. Asthma is a chronic inflammatory disease of the airways. Anti-inflammatory drugs constitute the mainstay of pharmacologic treatment. An especially important factor is patient education that encourages careful self-monitoring by using a peak flow meter. Environmental interventions are mandatory for all patients with asthma and may result in dramatic reductions in symptoms and medication use. Increased understanding of the pathophysiology of asthma has led to research in the development of "antimediator" agents, which may prove useful in the future. PMID- 7484717 TI - Proctalgia fugax. AB - Proctalgia fugax is a fairly common but little-known cause of rectal pain. It is a benign condition that has no known etiology. Symptoms consist of episodic, sudden, sharp pain in the anorectal area, usually lasting several seconds to several minutes. Diagnosis is based on a history that fits the classic picture, coupled with a normal physical examination. Several treatments have been tried and found anecdotally to be effective, although reassurance is the most useful therapeutic option. PMID- 7484718 TI - Management of patients with atypical and low-grade Pap smear abnormalities. AB - Family physicians frequently encounter patients with atypical and low-grade Papanicolaou (Pap) smear findings. While near consensus exists regarding the evaluation and management of patients with high-grade dysplasia and carcinoma detected on Pap smear, the appropriate management of patients with atypical and low-grade abnormalities continues to be controversial. Recent guidelines from the National Cancer Institute propose consideration of a more conservative management approach, with repeat Pap smear as an alternative to immediate colposcopy. Conservative management, as an alternative to an ablative or excisional procedure, has also been proposed for women with histologic evidence of lowgrade dysplasia. It is unlikely that an approach including a conservative diagnostic strategy and careful follow-up will result in an increase in the number of missed cases of cervical cancer. This assumption needs to be assessed by prospective clinical trials. PMID- 7484720 TI - Perioperative management of the alcohol-dependent patient. AB - Caring for the problem drinker in the perioperative period is a challenging task. If alcohol abuse is suspected, a careful assessment is indicated before surgery is performed. Both the CAGE and SMAST questionnaires are good screening tools for alcoholism. Preoperative evaluation of alcohol-dependent patients should include a complete blood count, blood urea nitrogen, serum electrolyte levels, creatinine and glucose levels, liver function tests, coagulation studies, an electrocardiogram and a chest radiograph. Smoking cessation and aggressive postoperative respiratory care are especially important for alcoholic patients who have chronic obstructive pulmonary disease. Elective surgery should not be considered in patients with acute hepatitis or cirrhosis, since the operative mortality rate is quite high in these patients. Alcohol withdrawal is managed primarily with benzodiazepines, although clonidine and beta blockers may also be beneficial. PMID- 7484719 TI - Cardiac rehabilitation as secondary prevention. Cardiac Rehabilitation Guideline Panel. PMID- 7484721 TI - Contemporary approaches to prenatal diagnosis. AB - A variety of options for prenatal diagnosis are available to the pregnant woman. Maternal serum analyte analysis, performed between 15 and 20 weeks' gestation, provides a screening test for fetal neural tube defects and aneuploidy in low risk pregnancies. Fetal ultrasound examination is of benefit in high-risk pregnancies. The necessity of prenatal screening in the low-risk patient with an established date of last menstrual period is more controversial. Ultrasound examination can establish gestational age, assess fetal number and position, determine placental location and amniotic fluid volume, and rule out major structural anomalies. Invasive fetal testing, including amniocentesis and chorionic villus sampling, should be offered to women who are 35 years of age or older or who have had abnormal results on noninvasive prenatal screening and in cases in which the parents are carriers of genetic conditions that are amenable to prenatal diagnosis. PMID- 7484723 TI - Management of behavior problems in the demented patient. AB - Behavior problems are common in persons with dementia and often lead to caregiver stress and institutionalization for the patient. In most cases, however, these problems are amenable to treatment. Although drug therapy may be necessary to manage some behavior problems, nonpharmacologic strategies may work as well, if not better, with fewer adverse effects. Support and education for the patient's family are the cornerstone of management. At some point, most demented patients display agitation. Appropriate, often nonpharmacologic management of agitation may include establishing a "no-fail" environment, limiting goals and providing reassurance. While delusions and hallucinations are also common, they seldom lead to agitation. Sleep disturbance and wandering are particularly upsetting to the patient's family, but these problems often respond to nonpharmacologic strategies, such as restricting naps and providing more cues about time and place. Depression, which occurs in many demented patients, is an especially treatable cause of disability. PMID- 7484722 TI - Glaucoma: early detection and therapy for prevention of vision loss. AB - Glaucoma is the second most common cause of permanent blindness in the United States and the most common cause of blindness among blacks. It is characterized by optic atrophy and vision loss, resulting from intraocular pressure that is too high to allow normal function of the optic nerve. Examination of the optic nerve and assessment of individual risk factors aids in the identification of patients who are at risk for glaucoma. Although glaucoma is a chronic disease, ocular complications can be prevented and sight may be preserved if the disease is detected early. A combination of antiglaucoma medications, laser treatment and filtration surgery is used to maintain intraocular pressure at a level sufficient to halt progression of optic nerve injury and prevent further vision loss. PMID- 7484724 TI - Interpreting thyroid tests. AB - Thyroid diseases are commonly encountered in family practice and are often suspected on the basis of the history and physical examination. The prevalence of thyroid disease in the general population is 0.5 percent, but it can be much higher in selected groups of patients, such as the elderly. A basic understanding of thyroid physiology and the methodology of laboratory thyroid testing is essential for the evaluation and management of patients suspected of having thyroid disease. Recently developed tests, such as the ultra-sensitive thyroid stimulating hormone test, provide the means for rapid and definitive diagnosis of common thyroid conditions. PMID- 7484725 TI - Oral nitrate preparations: an update. AB - Oral nitrates are used in the treatment of angina and congestive heart failure. In 1993, more than 12 million prescriptions for chronic administration of oral nitrates were written in the United States. Three nitrate compounds are currently available on the U.S. market: nitroglycerin, isosorbide dinitrate and the most recent, isosorbide mononitrate. Oral nitrates are available in sublingual, translingual (spray), transmucosal, chewable or sustained-release forms. Despite the complexity of choices, therapy can be simplified and remain efficacious, provided that attention is given to circumventing tolerance, a challenging problem with nitrate therapy. Tolerance can be avoided by ensuring that there is a daily nitrate-free interval using eccentric (i.e., 8 A.M. and 2 P.M.), twice daily dosing of standard or sustained-release forms, or frequent intermittent low dosages. PMID- 7484726 TI - The patient information and education initiative. PMID- 7484727 TI - ACOG issues technical bulletin on domestic violence. American College of Obstetricians and Gynecologists. AB - Physicians should attempt to identify individuals who are the victims of abuse and to help them understand the dynamics of their relationship and the dangers facing them and their children. Information about available community, social and legal resources, their legal rights and a plan for dealing with the abusive situation should be made available to these women. The physician must remain caring and supportive of the patient as she works through these crises, even if she chooses to follow advice or courses of action other than leaving the relationship. PMID- 7484728 TI - NIH develops consensus statement on infectious disease testing for blood transfusions. PMID- 7484729 TI - Gray hair, baldness, and wrinkles in relation to myocardial infarction: the Copenhagen City Heart Study. AB - To investigate a possible relation between aging signs such as graying of the hair, baldness, and facial wrinkling and myocardial infarction (MI), we analyzed data from The Copenhagen City Heart Study. During the 12-year follow-up, 750 cases of first-time MI were observed. After statistical adjustment for possible confounders, we found a correlation between graying of the hair, facial wrinkling, and frontoparietal baldness and crown-top baldness and MI in men. For example, the relative risk was 1.4 (95% CI, 0.9 to 2.0) for men with moderately gray hair compared with men with no gray hair and 1.9 (1.2 to 2.8) for men with completely gray hair (p < 0.001). With regard to gray hair, a similar although weaker and not statistically significant trend was seen in women. We conclude that, in addition to established coronary risk factors, aging signs like graying of the hair, male baldness, and facial wrinkling indicate an additional risk of MI. PMID- 7484730 TI - Short-term reproducibility of time domain, spectral temporal mapping, and spectral turbulence analysis of the signal-averaged electrocardiogram in normal subjects and patients with acute myocardial infarction. AB - The aim of this prospective study was to compare the short-term reproducibility of the signal-averaged ECG (SAECG) with three analysis techniques, conventional time-domain analysis, spectral-temporal mapping (STM), and spectral-turbulence analysis (STA), in a large series of normal subjects and patients with acute myocardial infarction (AMI). Two consecutive SAECGs were recorded in 225 consecutive patients 10.2 +/- 2.7 days after AMI and in 85 healthy volunteers. The visual, diagnostic, and quantitative reproducibility of the three techniques was compared. Time-domain analysis was the most reproducible method, having high R2 correlations, statistically fewer inconsistent diagnostic recordings, and statistically smaller differences compared with other techniques. STM was the least reproducible, justifying caution in its current form. Although STA was significantly less reproducible than time-domain analysis, it was also significantly better than STM. Two STA parameters, spectral entropy and interslice correlation mean, showed good reproducibility, suggesting that modification of this analysis technique could be useful in risk stratification. PMID- 7484732 TI - Relation between impedance and temperature during radiofrequency ablation of accessory pathways. AB - Temperature monitoring is a useful tool for radiofrequency ablation of accessory pathways. Impedance monitoring is also helpful, and an impedance fall of 20 ohm may predict coagulum formation. Therefore the purpose of this study was to prospectively quantitate the correlation between impedance and temperature during radiofrequency ablation. Thirty-three consecutive patients underwent radiofrequency ablation with a thermistor ablation catheter with continuous temperature and impedance monitoring during each energy application. The initial and final impedance and temperature measurements for 319 applications of radiofrequency energy were 109 +/- 17 ohm and 102 +/- 17 ohm (p < 0.0001), respectively, and 37 +/- 1 degree C and 57 degrees +/- 11 degrees C (p < 0.0001), respectively. Among the 319 applications of radiofrequency energy, 158 were associated with > or = 20 degrees C increase in tissue temperature, and 10 were associated with coagulum formation. A > or = 20 degrees C increase in tissue temperature was observed in 84 of 104 applications associated with a 5 to 10 ohm decrease in impedance (81% positive predictive value and 93% negative predictive value). The mean change in impedance for applications associated with and without coagulum formation was -19 +/- 7 ohm and -6 +/- 6 ohm (p < 0.0001), respectively, and coagulum formation never occurred with less than a 12 ohm decrease. In conclusions successful tissue heating without coagulum formation can be achieved by titrating the power to achieve a 5 to 10 ohm decrement in impedance. Impedance decrements beyond 10 ohm increase the likelihood of coagulum formation. PMID- 7484733 TI - Prospective study of retrograde coronary venography in patients with posteroseptal and left-sided accessory atrioventricular pathways. AB - The morphologic features of the coronary vein system was prospectively studied with retrograde venography in 117 patients with left-sided (78 patients) and posteroseptal accessory pathway (39). Findings were compared with accessory pathway localization. A mean of 3.3 +/- 1.5 venous branches draining into the coronary sinus or the great cardiac vein could be visualized. The morphologic condition was described and classified. Incidence, morphologic condition, and distribution did not differ between left-sided and posteroseptal accessory pathway. Venous abnormalities including ectasy, diverticulum, narrowing, angulation, and hypoplasia occurred in 22.2%. Diverticulum and narrowing were present in posteroseptal accessory pathway only and always related to the successful ablation site. In patients with left-sided accessory pathway, ectasy, angulation, and hypoplasia were found. Anomalies were less frequent (9% vs 43.6%, p < 0.001) and had no relation to accessory pathway localization. However, the successful ablation site was in 42.3% located < 5 mm to an angiographically visualized venous branch. In conclusion, posteroseptal accessory pathways are often related to coronary sinus abnormalities. In patients with a left-sided accessory pathway venous malformation is uncommon, whereas a close anatomic relation exists between accessory pathway localization and venous ventricular branches. PMID- 7484731 TI - Heart rate variability profiles in symptomatic coronary artery disease and preserved left ventricular function: relation to ventricular tachycardia and transient myocardial ischemia. Regression Growth Evaluation Statin Study (REGRESS). AB - We studied heart rate variability in ambulatory electrocardiographic recordings from 312 patients with significant coronary artery disease and a left ventricular ejection fraction of > or = 30%. Only those patients who had both episodes of ischemia and nonsustained ventricular arrhythmias showed diminished values for parasympathetic nonspectral and spectral heart rate (HR) variability parameters. The root mean square of difference of successive R-R intervals (RMSSD) and high frequency (HF) power in this group were 28 +/- 3 msec and 9 +/- 1 msec, respectively. The highest values of these HR variability parameters were found in patients with nonsustained ventricular tachycardia and no ischemia (81 +/- 21 msec and 14 +/- 1 msec, respectively, p < or = 0.05). A relation was seen between transient myocardial ischemia and low-frequency power and between transient ischemia and ventricular tachycardia. These data suggest that the autonomic nervous system may play a role in ischemia and in ischemia-related ventricular arrhythmias. PMID- 7484734 TI - Failures of epicardial and transvenous leads for implantable cardioverter defibrillators. AB - Lead failures constitute a major risk for patients with an implantable cardioverter defibrillator (ICD). However, data about the incidence and patterns of ICD-lead failures in a larger population are lacking. We analyzed the short term and midterm performance of 27 epicardial and 103 nonthoracotomy ICD-lead systems during a follow-up period of 36 +/- 21 months and 22 +/- 10 months, respectively (p < 0.05). The failure rate was 5 (19%) of 27 in the epicardial and 6 (6%) of 103 in the nonthoracotomy group (p < 0.05). The most common symptom was erroneous detection of ventricular fibrillation from artifact sensing in five patients. Two patients had to be resuscitated because of failure to defibrillate. Loss of pacing and loss of sensing were seen in two patients. Only two asymptomatic lead fractures could be diagnosed on routine radiograph. In conclusion, there was a considerable rate of lead failures, especially in epicardial systems. Long-term studies addressing the longevity of ICD leads, mechanisms of their failures, and improved diagnostic facilities are important to further increase the safety of this therapeutic approach. PMID- 7484735 TI - Asymmetric thickness of the left ventricular wall resulting from asynchronous electric activation: a study in dogs with ventricular pacing and in patients with left bundle branch block. AB - Various kinds of abnormal, asynchronous electric activation of the left ventricle (LV) decrease mechanical load in early versus late activated regions of the ventricular wall. Because myocardium usually adapts its mass to changes in workload, we investigated by echocardiography whether regional differences in wall thickness are present in two kinds of asynchronous electric activation of different origin and conduction pathway: epicardial ventricular pacing in dogs and left bundle branch block (LBBB) in patients. In six dogs, 3 months of epicardial LV pacing at physiologic heart rates decreased the thickness of the early activated anterior wall by 20.5 +/- 8.1% without significantly changing LV cavity area and septal thickness. In a retrospective study of 228 LBBB patients, the early activated septum was significantly thinner than the late activated posterior wall. The asymmetry most pronounced was as large as 10% in 28 patients with LBBB and paradoxic septal motion. No difference in regional wall thickness was present in 154 control patients. In conclusion, chronic asynchronous electric activation in the heart induces redistribution of cardiac mass. This redistribution occurs in hearts, which differ in impulse conduction pathway, disease, and species and is characterized by thinning of early versus late activated myocardium. PMID- 7484736 TI - Differing autonomic response to dobutamine in the presence and absence of ischemia: implications for the autonomic contribution to positive inotropic intervention. AB - Although it is well recognized that dobutamine is a powerful positive inotropic agent mediating increased myocardial contractility through direct beta-adrenergic stimulation, the mechanism of its overall circulatory effects appears more complex than that which may be ascribed solely to this direct action on the myocardium. Previous investigations have implied that reflex alterations in autonomic balance may contribute significantly to the response to this agent, but direct evidence describing such a response has not been reported. The objective of this investigation was to assess changes in autonomic tone induced by dobutamine in the presence and absence of ischemia through the measurement of heart rate variability in patients undergoing dobutamine stress echocardiography. Of 25 consecutive patients undergoing dobutamine stress echocardiography, 16 were found to have unequivocal evidence for or against the presence of ischemia during dobutamine infusion. Heart rate data from these 16 patients were submitted for spectral analysis of heart rate variability and quantification of parasympathetically governed high-frequency heart rate variability and sympathetically influenced low-frequency heart rate variability. Of the 16 patients nine were not found to have evidence for ischemia (group 1), and seven were found to have echocardiographic findings consistent with dobutamine-induced ischemia (group 2). The two groups significantly differed (p = 0.04) in the change in parasympathetic tone associated with dobutamine with a significant (p = 0.04) increase in parasympathetic tone in group 1 and a numeric decrease in group 2. A significant (p = 0.04) decrease in sympathetic tone was noted in group 1 as reflected by low-frequency heart rate variability with a numeric increase in this measure in group 2.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7484737 TI - Use of a tantalum-178 generator and a multiwire gamma camera to study the effect of the Mueller maneuver on left ventricular performance: comparison to hemodynamics and single photon emission computed tomography perfusion patterns. AB - During the Mueller maneuver, there is a decrease in intrathoracic pressure and an increase in transmural left ventricular pressure. The changes in loading conditions cause transient left ventricular dysfunction. This study examined the effects of the Mueller maneuver on left ventricular performance using tantalum (Ta)-178 (half-life 9.3 min) and a multiwire gamma camera. First-pass radionuclide angiograms were obtained at baseline and during Mueller maneuver in 41 patients aged 58 +/- 10 years. In 34 patients, stress single photon emission computed tomography (SPECT) myocardial perfusion imaging with thallium-201 or sestamibi was also performed. Hemodynamic measurements during the Mueller maneuver (n = 10) showed a decrease in systemic pressure (139 +/- 25 mm Hg vs 123 +/- 24 mm Hg, p < 0.001) and pulmonary artery pressure (24 +/- 6 mm Hg vs 14 +/- 12 mm Hg, p = 0.01) and an increase in heart rate (67 +/- 10 bpm vs 75 +/- 14 beats/min, p = 0.001). Among the 34 patients who had perfusion imaging, the left ventricular ejection fraction remained unchanged or increased in 17 patients (group 1) (48% +/- 19% vs 49% +/- 21%, p not significant) and decreased (> or = 5%) in 17 patients (group 2) (55% +/- 13% vs 40% +/- 16%, p = 0.001). The stress SPECT images showed no or only fixed defects in 11 (65%) patients in group 1 and 3 (18%) patients in group 2 (p = 0.02), and reversible defects in 6 (35%) patients in group 1 and 14 (82%) patients in group 2 (p = 0.04).(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7484738 TI - Influence of left ventricular mass on left ventricular diastolic filling in normotensive morbid obesity. AB - To identify factors influencing left ventricular (LV) diastolic filling in patients with morbid obesity, we performed transthoracic and Doppler echocardiography on 50 subjects whose actual body weight was > or = twice their ideal body weight and on 50 normal lean control subjects. The transmitral Doppler E/A ratio and E wave deceleration half-time were used to assess LV diastolic filling. Significant negative correlations were seen between the E/A ratio and the LV internal dimension in diastole (r = 0.819, p = 0.0001), systolic blood pressure (r = 0.751, p = 0.0001), LV end-systolic wall stress (r = 0.782, p = 0.0001), and LV mass/height index (r = 0.901, p = 0.0001). Significant positive correlations were seen between the E wave deceleration half-time and the LV internal dimension in diastole (r = 0.743, p = 0.0001), systolic blood pressure (r = 0.789, p = 0.0001), LV end-systolic wall stress (r = 0.828, p = 0.0001), and LV mass/height index (r = 0.831, p = 0.0001). No correlation was seen between diastolic blood pressure and either index of LV diastolic filling. Thus increasing LV mass is associated with progressive impairment of LV diastolic filling in morbidly obese individuals. The aforementioned alterations in LV loading conditions may contribute to impairment of LV diastolic filling directly or by increasing LV mass. PMID- 7484739 TI - Optimal determination of right ventricular filling dynamics in systemic hypertension. AB - To determine the optimal method of normalizing peak filling rate (PFR) determinations and apply it to the assessment of right ventricular (RV) and left ventricular (LV) filling characteristics and their interactions, 41 subjects with hypertension and 40 matched normals underwent echo-Doppler and nuclear study. Conventional normalization of PFR to end-diastolic volume (EDV) yielded poor correlations between nuclear- and echo-derived PFR (RV, r = 0.34; LV, r = 0.42), whereas nuclear and echo PFR normalized to stroke volume (SV) were closely correlated (RV, r = 0.87; LV, r = 0.92). Further, use of PFR normalized to SV revealed to close relation between RV and LV filling characteristics. Multivariate analysis confirmed that, in contrast to normalization to EDV or early to late filling-velocity ratios (E/A), peak filling rate normalized to SV was independent of ejection fraction and heart rate. In addition, RV filling impairment was related to LV filling impairment, and the effects of hypertension eliminated the independent influence of age on both LV and RV filling. In conclusion, normalization of PFR to SV may be preferable to use of EDV or E/A in evaluating RV and LV filling dynamics. PMID- 7484740 TI - Recurrent cerebrovascular events in patients with patent foramen ovale, atrial septal aneurysm, or both and cryptogenic stroke or transient ischemic attack. French Study Group on Patent Foramen Ovale and Atrial Septal Aneurysm. AB - Patent foramen ovale (PFO) and atrial septal aneurysm (ASA) have been identified as potential risk factors for stroke, but information about the risk of recurrent cerebral ischemia is scarce. The aim of this retrospective study was to assess the absolute risk of recurrent cerebrovascular events in 132 patients under 60 years of age with patent foramen ovale, atrial septal aneurysm (diagnosed by transesophageal echocardiography with a contrast study), or both and an otherwise unexplained stroke or transient ischemic attack (TIA). During a mean follow-up of 22.6 +/- 16 months, six patients had a recurrent stroke (n = 2) or a TIA (n = 4). No systemic embolism was observed. The actuarial risk of having a recurrent stroke was 2.3% (95% confidence interval, 0.6% to 8.2%) at 2 years, whereas the risk of having a stroke or a TIA was 6.7% (95% confidence interval, 3.1% to 14.2%) at 2 years. The average annual rates of recurrence were 1.2% and 3.4%, respectively. In patients with both PFO and ASA, the actuarial risk of a first recurrent stroke was 9.0% (95% confidence interval, 2.4% to 28.5%) at 2 years, with an average annual rate of recurrence of 4.4%. As a group, patients with patent foramen ovale, atrial septal aneurysm, or both and an otherwise unexplained stroke or TIA appear to have a low risk of recurrent stroke whatever the prophylactic antithrombotic therapy used. The association of ASA and PFO may be an indicator of a higher risk of recurrent stroke. PMID- 7484742 TI - Newer developments in the management of atrial fibrillation. AB - Atrial fibrillation (AF) is the most common sustained cardiac arrhythmia encountered in clinical practice. Unlike reentrant supraventricular tachycardia and malignant ventricular tachyarrhythmias, for which highly effective and safe nonpharmacologic therapies are available, the treatment of AF remains controversial and often problematic. Whereas electrical cardioversion restores sinus rhythm in most patients with AF, the maintenance of sinus rhythm often requires membrane-active antiarrhythmic drugs that may increase mortality by inducing ventricular proarrhythmia. The control of ventricular response rate, often associated with oral anticoagulation to prevent thromboembolic complications, is an alternative strategy in AF management. The relative efficacy and risks of these strategies and their respective role in different patient subgroups remain to be established. This article focuses on newer developments in the management of AF, including prospects for improved methods to maintain sinus rhythm, newer approaches to rate control, controversies regarding the use of oral anticoagulation, and novel nonpharmacologic therapies. These newer developments may lead over the next 10 years to a revolution in the management of AF as profound as that produced over the last 10 years by nonpharmacologic therapy of other arrhythmias. PMID- 7484741 TI - Angiotensin-converting enzyme gene polymorphism in Japanese patients with hypertrophic cardiomyopathy. AB - To examine the contribution of the angiotensin-converting enzyme (ACE) gene to hypertrophic cardiomyopathy (HCM), we determined the ACE insertion/deletion (I/D) polymorphism in 80 patients with HCM and 88 of their unaffected siblings and children. Patients were divided into familial or solitary HCM (FHCM or SHCM) groups with or without affected family members. Genotypes were identified by the polymerase chain reaction (PCR) with oligonucleotide primers flanking the polymorphic region in intron 16 of the ACE gene to amplify template DNA prepared from peripheral leukocytes. D-allele frequencies were 0.38 in all subjects, 0.42 in patients with HCM, and 0.35 in relatives (p < 0.05). The probability ratios were 1.98, 1.46, and 2.97 in patients with HCM, FHCM, and SHCM, respectively. The D allele frequency was higher in SHCM than in FHCM (p < 0.05). The findings suggest that HCM, especially in solitary cases, is partially determined by genetic disposition. Findings imply that the ACE D allele is one of the genetic contributing factors associated with cardiac hypertrophy in HCM. PMID- 7484744 TI - Observation of atherosclerotic lesions by an intravascular microscope in patients with arteriosclerosis obliterans. AB - The magnification power of conventional fiberscopes for intravascular use is up to x30, and therefore they are not feasible for observation of fine structures of vascular changes. We devised a microscope that enables percutaneous transluminal observation of the vascular luminal changes of cellular order. Thus we examined its feasibility in patients. The microscope was 8F in diameter with a rod lens inside, and its magnification power was from x150 to x350. The microscope was introduced into the iliac or femoral artery for observation of vascular lesions during angioplasty in seven patients with arteriosclerosis obliterans. With the aid of vital staining individually damaged endothelial cells, fibrin threads, foam cells, collagen, elastic fibers, and structure of microthrombi were discriminated clearly. The results indicate that structures of human vascular lesions of cellular order can be observed percutaneously by the intravascular microscope. PMID- 7484743 TI - Utility of the 12-lead electrocardiogram in localizing the origin of right ventricular outflow tract tachycardia. AB - The purpose of this investigation was to develop an algorithm on the basis of the QRS morphology observed on the 12-lead ECG that would rapidly locate the site of origin of the monomorphic ventricular tachycardia arising from the septal portion of the RVOT. Radiofrequency catheter ablation guided by pace-mapping techniques has proven effective in eliminating the ventricular tachycardia originating from the RVOT in the absence of structural heart disease. A method that would rapidly identify the portion of the RVOT septum toward which more detailed pace-mapping should be directed before catheter ablation would be useful in decreasing procedure time and radiation exposure and potentially facilitating a successful ablation procedure. The RVOT septum was divided into nine sites. In 11 patients, bipolar pacing was performed at each of the nine designated sites to mimic ventricular tachycardia. A standard 12-lead surface ECG was recorded during pacing. The QRS morphology in the limb leads was characterized and the site of the R-wave transition was determined in the precordial leads. A QS in lead a VR and a monophasic R wave in leads II, III, aVF, and V6 were noted in each patient at all paced sites. In lead I, pacing at the three posterior septal sites always resulted in an R wave. Pacing at the three anterior sites produced a dominant Q wave (either QS or Qr) at 17 (52%) of 33 sites or a qR complex at 16 (48%) of 33 sites.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7484745 TI - Acute stent thrombosis associated with exercise testing after successful percutaneous transluminal coronary angioplasty. PMID- 7484746 TI - Takayasu's arteritis with unstable angina and aortic insufficiency. PMID- 7484747 TI - Acquired coronary artery fistula in children after ventricular septal myectomy of the right or left ventricular outflow tract. PMID- 7484748 TI - Right atrial mass with attachment to the atrial septum caused by metastatic renal cell carcinoma. PMID- 7484749 TI - Pheochromocytoma with catecholamine-induced impairment of cardiac neuronal function. PMID- 7484751 TI - Matching to guide the design and predict the outcome of randomized atherectomy trials. PMID- 7484752 TI - Evolution of the management of malignant ventricular tachyarrhythmias: the roles of drug therapy and implantable defibrillators. PMID- 7484750 TI - Infants with long-QT syndrome and 2:1 atrioventricular block. PMID- 7484757 TI - Directional coronary atherectomy and progressive coronary dilatation: a comparative analysis of acute outcome. AB - To evaluate the acute results and in-hospital complications of directional atherectomy (DCA) as compared to progressive coronary dilatation (PCD), we retrospectively analyzed the acute outcome of DCA with PCD in age-, sex-, vessel , and lesion morphology-matched groups of patients during the same time span. There was a total of 73 matched patients (77 lesions) in each group. Angiographic success on the basis of intent to treat was 85% in the DCA cohort versus 97%. The preprocedural mean diameter stenosis was similar between the two groups (87% vs 84%; p = n.s.). The mean postprocedural stenosis was significantly lower with DCA than with PCD (11.2% vs 19.7%; p < or = 0.05). Complications including death, myocardial infarction, and need for emergency bypass surgery were not statistically different in either group. In conclusion, PCD offers an alternative method of coronary intervention in patients with "atherectomy anatomy" with a significantly higher success rate. It can also be used successfully when DCA fails or cannot be performed because of technical factors. PMID- 7484753 TI - Serial changes of plasma plasminogen activator inhibitor activity in acute myocardial infarction: difference between thrombolytic therapy and direct coronary angioplasty. AB - The fibrinolytic system is impaired in patients with acute myocardial infarction (AMI). The primary regulatory element of fibrinolytic activity is plasminogen activator inhibitor (PAI). There are no reports, however, on the serial changes of PAI activity after thrombolysis or coronary angioplasty in patients with AMI undergoing emergency coronary angiography. This study was designed to examine the difference in the change of fibrinolytic activity between patients with AMI who underwent thrombolytic therapy with recombinant tissue-plasminogen activator (rTPA) and those who underwent direct percutaneous coronary angioplasty (PTCA). We measured the serial changes of PAI activity and tissue plasminogen activator (TPA) antigen after rTPA therapy or direct PTCA. Twenty-two patients received emergency coronary angiography and were treated with rTPA intravenously. Twenty patients underwent direct PTCA. Plasma PAI activity levels were increased on admission and further increased within 24 hours in patients treated with rTPA and in those treated with direct PTCA. In the thrombolysis group, there were two peaks in plasma PAI activity levels (IU/ml) at 4 hours (27.0 +/- 2.9) and at 16 hours (25.6 +/- 2.5) after the initiation of rTPA infusion. However, in the direct PTCA group, there was one peak of PAI activity (IU/ml) at 16 hours (23.9 +/- 2.7) after the initiation of direct PTCA. In conclusion, the PAI activity has two peaks in the thrombolysis group and one peak in the direct PTCA group. PMID- 7484755 TI - Noninvasive assessment of no-reflow phenomenon in a canine model of reperfused infarction by contrast-enhanced magnetic resonance imaging. AB - The aim of this study was to test whether contrast-enhanced magnetic resonance (MR) imaging may assess in vivo the severity of the no-reflow phenomenon in a dog model of infarction with 2-hour coronary occlusion followed by reperfusion (6 hours). Subsecond MR imaging combined with intravenous bolus administration of superparamagnetic iron oxide particles (SPIO) was performed at the fifth hour of reperfusion. An MR index was calculated using the difference of signal-intensity enhancement between ischemic and nonischemic zones during the SPIO first pass. Dogs were separated into two groups according to the severity of ischemia: collateral blood flow in the central ischemic zone at 120 minutes of occlusion (radioactive microsphere technique) < 22.5% of the flow in the nonischemic zone (group I) or > 22.5% (group II). Mean collateral blood flow during occlusion was lower in group I (11.3% +/- 2.9%, n = 7) than in group II (66.8% +/- 19.8%, n = 6, p < 0.05). Mean infarct size was significantly larger in group I (58.6% +/- 4.9% of the area-at-risk, n = 7) than in group II (16.5% +/- 6.5%, n = 6, p < 0.05). For the entire population (n = 13), the infarct size was inversely correlated to the collateral blood flow (r = -0.64, p = 0.02, standard error of estimate = 0.24). The relative rate of enhancement in ischemic myocardium (MR index) was significantly lower in group I (38.1% +/- 10.9%) than in group II (142.8% +/- 32%; p < 0.05).(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7484756 TI - Effect of 100% oxygen administration on infarct size and left ventricular function in a canine model of myocardial infarction and reperfusion. AB - High oxygen concentrations reduced infarct size in prereperfusion era studies; however, with reperfusion therapy, high oxygen tension carries the theoretical risk of exacerbating reperfusion injury by increasing toxic oxygen-derived free radicals. In this study, two groups of dogs underwent 90 minutes of coronary occlusion and 72 hours of reperfusion. The oxygen group (n = 16) received 100% inspired oxygen from 20 minutes before reperfusion through 3 hours of reperfusion, whereas the room-air group (n = 19) was ventilated with room air. Infarct size (as a percentage of risk area) was reduced by 38% in the oxygen group (26.7% +/- 4.7% vs 43.3% +/- 4.3%; p = 0.017). This benefit was independent of underlying variability in collateral blood flow in individual dogs (p = 0.016 by analysis of covariance [ANCOVA]). Left ventricular ejection fraction was significantly improved in the oxygen group (43% +/- 3% vs 33% +/- 2%; p = 0.008), as was regional function in the infarct zone (p < 0.05). These data suggest that high concentrations of inspired oxygen may also benefit patients with acute myocardial infarction who undergo reperfusion therapy. PMID- 7484758 TI - Influence of plaque composition on luminal gain after balloon angioplasty, directional atherectomy, and coronary stenting. AB - This study was conducted to correlate the acute luminal enlargement achieved by three different nonsurgical revascularization procedures in 79 patients (32 treated by balloon angioplasty, 29 by directional atherectomy, and 18 by coronary stenting) with the morphologic characteristics of coronary plaques assessed by preprocedure intravascular ultrasound. The absolute luminal gain was 2.41 +/- 1.54 mm2 for balloon angioplasty, 3.17 +/- 1.8 mm2 for directional atherectomy, and 4.56 +/- 1.45 mm2 for coronary stenting (p = 0.00005). However, when luminal gain was corrected for the external vessel area (luminal gain index), such difference was no longer present (0.22 +/- 0.12 for balloon angioplasty, 0.24 +/- 0.15 for directional atherectomy, and 0.30 +/- 0.12 for coronary stenting, p = not significant). Concentric plaques treated by coronary stenting had a higher luminal gain index than eccentric plaques (p = 0.01). A comparison of the three devices showed that a similar luminal gain index was achieved in soft plaques, whereas coronary stenting was superior to directional atherectomy (0.41 +/- 0.10 vs 0.20 +/- 0.09, p = 0.002) and balloon angioplasty (0.41 +/- 0.10 vs 0.19 +/- 0.08, p = 0.0005) in concentric plaques. Coronary stenting also induced a greater luminal gain index than directional atherectomy in calcific plaques (0.30 +/- 0.11 vs 0.18 +/- 0.09, p = 0.04). In conclusion, these data show that plaque morphology assessed by preprocedure intracoronary ultrasound influences the acute luminal enlargement achieved by different coronary interventions. The knowledge of plaque composition may be useful in guiding the choice of the device to be used to obtain a larger acute luminal gain. PMID- 7484754 TI - Effect of the hydrophilic alpha-tocopherol analog MDL 74,405 on detection of hydroxyl radicals in stunned myocardium in dogs. AB - We have previously shown in dogs that the hydrophilic alpha-tocopherol analog, MDL 74,405, attenuates postischemic myocardial dysfunction ("stunning") and generation of free radicals as assessed with the spin trap alpha-phenyl N-tert butyl nitrone (PBN). However, we could not discern whether this drug acts on primary radicals (such as hydroxyl radical [.OH]) or on secondary radicals. The goal of this study was to directly determine whether the beneficial effects of MDL 74,405 result from actions against .OH. Open-chest dogs undergoing a 15 minute coronary artery occlusion and 3 hours of reperfusion received an intravenous infusion of either saline solution (control group, n = 7) or MDL 74,405 (n = 6) starting 30 minutes before coronary occlusion and ending 60 minutes after reflow at a dose of 0.3 mg/kg/hr. Formation of .OH was estimated by the technique of aromatic hydroxylation of phenylalanine. Phenylalanine was infused intravenously, and the plasma concentrations of the hydroxylated products ortho-, meta-, and para-tyrosines (o-, m-, and p-tyr) in the coronary venous effluent and in the arterial blood were measured with high-performance liquid chromatography. In the control group a dramatic increase in the myocardial release of o-, m-, and p-tyr was observed immediately after reperfusion; the release of tyrosines peaked at 1 minute of reflow and continued up to 10 minutes after reperfusion. MDL 74,405 abolished the release of o-tyr throughout the first 10 minutes of reperfusion but had a less pronounced effect on the production of m and p-tyr. These results demonstrate that MDL 74,405 is effective in inhibiting .OH-initiated reactions in the postischemic stunned myocardium in the dog, suggesting that the anti-.OH action of MDL 74,405 is an important mechanism of action of this antioxidant. PMID- 7484759 TI - Intravascular ultrasound appearance of normal and mildly diseased coronary arteries: correlation with histologic specimens. AB - Intravascular ultrasound imaging was performed in vitro on six histologically normal and 104 minimally diseased arteries in patients aged 13 to 83 years. This study tested the hypothesis that normal coronary arteries produce a three-layer image that corresponds to the histologic layers of intima, media, and adventitia. The results showed a very good correlation between area of the echolucent ultrasound layer with the media and the inner echogenic layer with intimal area. In addition, a three-layer appearance was consistently seen when the internal elastic membrane was present with or without intimal hyperplasia. If the internal elastic membrane was absent, a three-layer appearance was still seen if the collagen content of the media was low. However, a two-layer appearance was observed when there was absence of the internal elastic membrane as well as a high collagen content of the media. PMID- 7484760 TI - Findings of coronary angioscopy in angiographically normal coronary segments of patients with coronary artery disease. AB - Compared with pathologic studies coronary angiography is a relatively insensitive technique to detect early atherosclerosis. Coronary angioscopy is a new technique providing direct information on luminal vessel surface. To determine whether coronary angioscopy may detect the presence of atherosclerotic disease on angiographically normal coronary segments, 52 patients underwent a study with coronary angioscopy before coronary angioplasty. The mean age was 59 +/- 10 years; 46 patients were men and 6 were women. The reason for coronary angioplasty was unstable angina in 36 patients, stable angina in 8 patients, and silent ischemia in 8 patients. In seven patients angiography revealed luminal irregularities on the coronary segment proximal to the culprit lesion, and all these patients also had proximal disease as demonstrated by coronary angioscopy. In the remaining 45 (87%) patients angiography revealed a smooth-vessel contour proximal to the target lesion. On quantitative angiography these "normal" coronary segments measured 2.8 +/- 0.4 mm in luminal diameter. In 30 (67%) of these patients angioscopy revealed proximal disease on the vessel wall, but in 15 (33%) patients the luminal surface of these segments also appeared normal on angioscopy. Disease as detected by angioscopy in angiographically normal segments included yellow plaque in 19 patients, mural thrombus in 5, mixed plaques in 4, and small flaps in 2 patients. In eight patients coronary angioscopy detected that atherosclerotic disease extended proximally from the target lesion, but in the remaining 22 patients the angioscopic findings appeared to be discrete and well separated from the angiographic lesion. All these plaques were relatively small and did not protrude into the coronary lumen.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7484762 TI - Serial assessment of left ventricular function after myocardial infarction. AB - Left ventricular (LV) function is an important predictor of morbidity and mortality after myocardial infarction (MI). Changes in LV function have been examined during the early and late phases after MI, but serial measurements of LV function during the subacute period have not been performed. To assess sequential changes in LV function during the subacute period after MI, we used quantitative two-dimensional echocardiography to examine 22 patients over a 1-year period. Twenty-one of the 22 patients had a Q-wave MI. Eleven had an anterior MI and 10 had an inferior MI; their peak creatine phosphokinase (CPK) was 1213 mIU/ml +/- 14. Three weeks after acute MI, LV ejection fraction (LVEF) had increased from 45% to 52%. Seven of 19 patients showed an LVEF < 43% at baseline. In five of these patients, LVEF improved, but in two patients, LVEF was still < 43% in week 3. There was a significant enlargement of LV end-diastolic volume (LVEDV) (94 ml to 112 ml, p < 0.05) across the four observations but no change in LV end systolic volume (LVESV; 54 ml to 56 ml, p = n.s.). When two groups (G1 [depressed], LVEF < or = 43%; G2 [preserved], LVEF > 43%) were compared, the group with depressed LVEF demonstrated a higher probability of improvement in LVEF (34% to 47%, p < 0.001) and stroke volume (38 ml to 65 ml, p < 0.01). PMID- 7484761 TI - Assessing myocardial viability: correlation of myocardial wall motion abnormalities and pathologic Q waves with technetium 99m sestamibi single photon emission computed tomography. AB - The clinical benefit of coronary revascularization depends largely on the viability of the myocardium that is perfused. To determine if the combination of electrocardiogram and left ventriculography findings could be used to predict viability, the presence of pathologic Q waves and wall motion abnormalities on contrast left ventriculography were correlated with findings on stress sestamibi scanning in 201 patients. Wall motion was abnormal in 51.5% of 103 Q regions; 30 (56.6%) of these had fixed sestamibi defects, and 22.6% had fully or partially reversible sestamibi defects. Q waves were associated with 43.4% of 122 regions with wall motion abnormality; 67.9% of these areas had fixed or partially fixed sestamibi defects. Wall motion abnormalities were present in 46.1% of 104 areas with fixed sestamibi defects. Although there was a statistically significant correlation among Q waves, left ventricular wall motion abnormalities, and stress sestamibi uptake (and various combinations of these data), the relatively large number (53.8%) of discordant findings (e.g., normal ventricular wall motion in the presence of fixed sestamibi defects) suggests that nonviability cannot be assumed without at least assessing both contractile left ventricular motion and metabolic (e.g., sestamibi scanning) function. PMID- 7484763 TI - Changes in patient treatment after abrupt closure complicating percutaneous transluminal coronary angioplasty: a historic perspective. AB - This study compares the incidence and management of acute closure complicating coronary angioplasty in three historic populations of patients having undergone the procedure at the same center: group 1 (n = 146 of 881) ("early years" of angioplasty, 1980 to 1986), group 2 (n = 113 of 1781) (bailout stenting learning curve, 1990 to 1992), and group 3 (n = 34 of 525) (1993). The incidence of acute closure decreased from group 1 (146 [17%] of 881) to groups 2 and 3 (147 [6%] of 2306); (p < 0.001). Management of the occlusion changed over the years, with less emergency coronary bypass surgery ([36%] 52 of 146, 15 [13%] 113, and 3 [9%] of 34), respectively, p < 0.01) and more repeat angioplasty (70 [48%] of 146; 87 [78%], of 113, and 30 [88%] of 34, p < 0.001). The use of prolonged inflations (> 10 minutes) and stenting increased from group 2 (15 [13%] of 113 and 16 [14%] of 113, respectively) to group 3 (12 [35%] of 34, and 10 [30%] of 34, respectively). In-hospital death occurred in 18 (12%) of 146, 7 (6%) of 113), and (2 (6%) of 34) patients in the three groups. Acute myocardial infarction decreased from 64% to 46% and 27%, respectively (p < 0.01). Overall, the number of patients free of events at hospital discharge increased from 38 (26%) of 146 to 53 (47%) of 113 (p < 0.001) and to 23 (68%) of 34 (p < 0.001).(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7484764 TI - Unplanned use of intracoronary stents for the treatment of a suboptimal angiographic result after conventional balloon angioplasty. AB - This observational single-center trial examines the safety and efficacy of unplanned endoluminal stenting for the treatment of a suboptimal angiographic result (defined as a residual stenosis after angioplasty of 40% to 50% without delayed runoff as estimated by visual assessment) after conventional coronary angioplasty in native, new-onset, coronary artery stenoses. Between October 1991 and April 1994, 101 patients with suboptimal results after coronary angioplasty in new-onset lesions were treated by endoluminal Wiktor (41 patients) and Palmaz Schatz (60 patients) stent implantation. Stenting was a technical and angiographic success in all cases. In-hospital complications were subacute closure (2%) and vascular complications at puncture site necessitating surgery (12%) or blood transfusion (3%). No myocardial infarction occurred, nor was any urgent bypass surgery performed. At follow-up restenosis was detected in 16 (20%, 80% angiographic follow-up rate) patients requiring repeat angioplasty (8%) and elective bypass grafting (4%). Myocardial infarction was not documented. However, one patient died suddenly at 5 months of follow-up. The unplanned use of intracoronary stents is a safe and effective therapeutic option for the treatment of a suboptimal angiographic result after conventional angioplasty in new-onset lesions. This approach guarantees a high immediate angiographic success but implies a considerable incidence of vascular complications at puncture site. PMID- 7484765 TI - Risk factors for restenosis after percutaneous transluminal coronary angioplasty: role of lipoprotein (a). AB - To evaluate serum levels of lipoprotein (a) (Lp[a]) as a predictor of restenosis after percutaneous transluminal coronary angioplasty (PTCA), we evaluated 71 patients who underwent elective single-vessel angioplasty. Patients were divided into two groups according to the presence (n = 24 [34%]; group R) or absence (n = 47 [66%]; group N) of restenosis. Serum insulin levels were similar before and after the glucose challenge test in both groups. The median level of serum Lp(a) was 34.9 mg/dl in group R compared with 19.4 mg/dl in group N (p < 0.01). The frequency of the apo E4 allele was 4 (17%) in group R and 4 (9%) in group N (p = NS). The incidence of restenosis was significantly higher in patients with Lp(a) levels > or = 30 mg/dl than in those with Lp(a) levels < 30 mg/dl (65% vs 26%; p < 0.01). Our results indicate that a serum Lp(a) level > or = 30 mg/dl is a risk factor for restenosis. PMID- 7484766 TI - Comparison of local intravascular drug-delivery catheter systems. AB - Systemic and local delivery of the photosensitive drug Photofrin polyporphyrin was investigated in normal porcine arteries (n = 192). A macroporous balloon and a novel needle injection catheter were used for local drug delivery and compared with systemic delivery. Fluorescence microscopy combined with digital image analysis was used to quantify the drug-related fluorescence. Systemic delivery showed a maximum in the intima at 4 hours. Application with the porous balloon revealed maximum indicator-related fluorescence intensity in the intima after 5 minutes; delivery with the needle injection catheter resulted in a several-fold enhanced maximum in adventitia after 30 minutes compared with the maximum achieved with either systemic injection or porous balloon application. After 21 days fluorescence was detectable in arteries treated with the new needle injection catheter. Local drug delivery is feasible with either system, but prolonged delivery was achieved only with the needle injection catheter. PMID- 7484767 TI - Angiogenic therapy of acute myocardial infarction by intrapericardial injection of basic fibroblast growth factor and heparin sulfate: an experimental study. AB - To examine whether angiogenesis and myocardial salvage occur, 30 micrograms basic fibroblast growth factor (bFGF) and 3 mg heparin sulfate (HS) were injected through the right atrium into the pericardial cavity by a thin needle-tipped catheter in a canine model of acute myocardial infarction. One month later infarcted weight/left ventricle weight was 24% +/- 5.2%, 25% +/- 4.0%, 18% +/- 2.4%, and 10% +/- 1.8% (mean + SE) in saline solution, HS, bFGF alone, and bFGF plus HS groups, respectively. Vascular number in the infarcted area of the outer layer was 13 +/- 3.3, 20 +/- 2.2, 47 +/- 8.3, and 136 +/- 26.3/200 x 200 microns2 in saline solution, HS, bFGF alone, and bFGF plus HS groups, respectively. Thus the vascular number was the largest in the bFGF plus HS group. The vascular number was larger in the subepicardial than in the subendocardial infarcted areas. Vessels directed from the epicardium toward the subepicardial infarcted area were also observed. The transcatheter intrapericardial injection of bFGF plus HS caused angiogenesis and myocardial salvage. This method might bring about a selective therapeutic and preventive modality of myocardial infarction irrespective of coronary anatomy and contraindications for coronary interventions and surgery. PMID- 7484768 TI - Temporal variability in wall motion abnormality with stress-induced myocardial ischemia assessed by quantitative exercise echocardiography: difference between reversible and persistent myocardial ischemia. AB - Echocardiography was performed before and after treadmill tests in 30 patients to determine whether there is temporal variability in left ventricular wall motion abnormality with exercise-induced ischemia. The extent of wall motion was assessed by calculating systolic shortening in endocardial motion by two methods: (1) comparison between end diastole and end systole (ED-ES) and (2) frame-by frame analysis throughout systole (integration). The extent of myocardial ischemia was assessed by stress thallium 201 emission tomography. Sensitivity of echocardiography for diagnosis of ischemia was 62% by ED-ES, but it increased to 83% by the integration method. Twenty-one percent of the ischemic area showed wall motion abnormality solely in the middle of systolic phase but not at end systole, and this occurred more at the reversible ischemic area (31%) than at the persistent ischemic area (19%). In conclusion, analysis throughout systole is important in detecting a reversible ischemic area. PMID- 7484769 TI - Use of positron emission tomography for prediction of perioperative and late cardiac events before vascular surgery. AB - The efficacy of myocardial perfusion imaging for cardiac-risk stratification of patients undergoing vascular surgery has been disputed recently. In comparison with conventional techniques, positron emission tomography (PET) has the benefit of permitting a true resting scan, allows accurate measurement of the extent of ischemia, and is highly specific for the diagnosis of coronary disease. We therefore investigated the use of PET for risk stratification at the time of vascular surgery and subsequent follow-up in 78 patients (aged 67 +/- 11 years, 52 men), selected for testing before the performance of extensive surgery or because of one or more clinical risk factors. Perfusion images were obtained by using a standard rubidium 82 protocol before and after dipyridamole-handgrip stress. With use of a quantitative color scale in a 24-segment model of the left ventricle, scans were reported as showing normal perfusion, resting defects, or stress-induced defects (deterioration > 15% with stress). After exclusion of 6 patients referred for myocardial revascularization, 72 patients were followed up in the perioperative period and for 18 +/- 12 months for late cardiac death, myocardial infarction, or unstable angina. Perioperative events occurred in 14 patients (5 with myocardial infarction and 9 with unstable angina), 10 of whom had ischemia at PET (sensitivity, 71%; predictive value of a positive test, 45%). Isolated resting perfusion defects were not associated with events. The presence of extensive ischemia (more than five segments) had a positive predictive value of 64%, and its absence gave a negative predictive value of 89%.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7484770 TI - Alterations in heart rate variability in patients undergoing dobutamine stress echocardiography, including patients with neurocardiogenic hypotension. AB - Frequency-domain analysis of heart rate variability (HRV) appears to distinguish sympathetic and parasympathetic modulation of heart rate. The effects of acute reflex, as opposed to tonically augmented, cardiac vagal tone on HRV are not clearly defined. Power spectral components of HRV were measured in 36 patients undergoing dobutamine stress echocardiography including during episodes of neurocardiogenically mediated hypotension. The low-frequency (LF; 0.04 to 0.15 Hz) and high-frequency (HF; 0.15 to 0.40 Hz) components of HRV decreased with increasing dose of dobutamine (5 to 40 micrograms/kg/min) in patients whose systolic blood pressure, compared with baseline, remained within 20 mm Hg (normotensive) or increased > or = 20 mm Hg (hypertensive). The ratio of LF/HF became < 1 in the hypertensive group at 30 and 40 micrograms/kg/min of dobutamine. In eight patients in whom apparent neurocardiogenically mediated hypotension developed (decrease in systolic blood pressure > or = 20 mm Hg from baseline), LF and HF measures became significantly higher than those in the patients without hypotension, whereas the LF/HF ratio was unchanged. These findings suggest that the HF component of HRV is a reliable measure of reflex augmented cardiac vagal activity and lend further support to the importance of parasympathetic modulation of the LF component of HRV. PMID- 7484771 TI - Upright postures and isoproterenol infusion for provocation of neurocardiogenic syncope: a comparison of standing and head-up tilting. AB - Head-up tilt testing has proved to be useful in provocation of neurocardiogenic syncope. The purpose of this study was to examine whether simply assuming an upright posture by standing can be an alternative to the head-up tilt testing for diagnosis of neurocardiogenic syncope. Eighty-four patients with recurrent unexplained syncope and 22 normal volunteers were recruited into the study. Forty seven patients with syncope and all normal volunteers received the standing test. Thirty-seven of the patients with syncope received head-up tilt testing (90 degrees). All subjects lay down for 5 minutes and then assumed an upright posture until syncope or presyncope occurred or until a maximum of 10 minutes was reached in each stage of the test. The tests included four stages: baseline and infusion of 1, 2, or 3 micrograms/min isoproterenol in each of the successive stages. Five subjects could not tolerate the procedure, and further testing was terminated. Overall, the standing test was positive in 83% of the patients with syncope, and its specificity was 74%. The head-up tilt testing was positive in 75% of the patients with syncope. The duration of assuming an upright posture before occurrence of syncope or presyncope was significantly longer in the syncope tilting group in the third stage (p < 0.01) and the fourth stage (p < 0.05) compared with the syncope-standing group. However, the curves of the time course for cumulative positive rates were not significantly different (p = 0.0739) in the two groups.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7484773 TI - Transesophageal versus transthoracic echocardiography in the evaluation of right atrial tumors. AB - The purpose of this study was to define the contribution of transesophageal echocardiography to the diagnosis of right atrial tumors in a large series of patients with this rare finding. Transesophageal echocardiography (TEE) has been found to be valuable in evaluating patients with intracardiac masses and has been shown to be superior to transthoracic echocardiography (TTE) in evaluating left heart masses. Between 1989 and 1993, 23 patients with either known tumors elsewhere or right atrial masses that were detected on TTE were evaluated with TEE. TEE studies were performed in the noninvasive cardiology laboratory. All patients were studied with 5 MHz biplane or omniplane transducers. The right atrium was evaluated for the presence, characteristics, extent, and attachment of any masses and for extension of these masses into the great vessels or other cardiac chambers. No complications occurred. Six patients had primary right atrial tumors. In 10 patients the tumors reached the right atrium via the inferior vena cava. Seven patients had malignant secondary tumors. TEE demonstrated three tumors not detected by TTE. Furthermore TEE provided 16 additional findings not seen on TTE. In conclusion, TEE is superior to TTE in the evaluation of right atrial tumors. TEE should be considered in patients with right atrial tumors even when these tumors have been demonstrated with TTE. PMID- 7484772 TI - Tamponade in patients undergoing cardiac surgery: a clinical-echocardiographic diagnosis. AB - The purpose of this study was to evaluate the sensitivity of current echocardiographic criteria in detecting cardiac tamponade in the patient who has undergone cardiovascular surgery. Because the current echocardiographic criteria for tamponade were initially developed and studied predominantly in patients with medical problems, relatively less information is available in patients who have undergone cardiac surgery. Of 848 consecutive patients who underwent cardiovascular surgery, patients were selected for the study if they had clinical or hemodynamic deterioration and had undergone an echocardiogram just before a successful pericardiocentesis or a surgical evacuation of pericardial blood or clot. The echocardiograms were evaluated for evidence of chamber collapse, cardiac motion, Doppler flow variations, and the location and width of pericardial separation. Fourteen patients were identified who met the inclusion criteria (clinical or hemodynamic deterioration, recent echocardiogram, and successful intervention) for cardiac tamponade. The clinical and hemodynamic findings were hypotension (13 patients), low cardiac output (7), low urine output (3), cardiopulmonary arrest (1), elevated central venous pressure (1), and shortness of breath (1). In these patients current echocardiographic criteria were seen infrequently: chamber collapse in the right atrium (6 of 14 patients) and right ventricle (4 of 14); Doppler flow variation (2 of 5); and swinging heart (0 of 15), whereas increased pericardial separation (> or = 10 mm) was seen in all (14 of 14) the patients.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7484775 TI - Multiplane transesophageal echocardiography in the diagnosis of congenital coronary artery fistula. AB - The purpose of this study was to examine the advantages of multiplane transesophageal echocardiography in the diagnosis of congenital coronary artery fistula, specifically in depicting the origin, the course, and the drainage site. Seven consecutive patients ranging in age from 20 to 72 years with a suspected coronary artery fistula underwent conventional transthoracic and multiplane transesophageal echocardiographic studies between March 1993 and July 1994. When a coronary artery fistula was noted, the origin, the course, and the drainage site were carefully searched for. All patients then underwent a cardiac catheterization with the performance of coronary angiography. A large right coronary artery fistula was detected in three patients; one of them had a drainage to the posterior left ventricle, one to the lateral right ventricle, and the other to the medial aspect of the right ventricle just below the insertion of the septal leaflet of the tricuspid valve. A small coronary artery fistula arising from the left coronary artery was noted in four patients, two from the left anterior descending artery and the other two from the left circumflex artery. Three of these four patients had a drainage to the main pulmonary artery and one to the left ventricle. The drainage site was clearly depicted in all seven patients, whereas the origin and the course were precisely defined in five patients by using multiplane transesophageal echocardiographic examination. The multiplane transesophageal echocardiography provides a panoramic view of the coronary artery and the fistulous vessel with a precise definition of the origin, the course, and the drainage site of the fistula. Therefore it is the noninvasive diagnostic mode of choice. PMID- 7484774 TI - Quantitation of echocardiographic predictors of outcome in infants with isolated ventricular septal defect. AB - Clinical and morphometric features such as ventricular septal defect (VSD) size and location may determine outcome in infants with an isolated VSD. However, no currently available data allow quantitative estimation of the probability of spontaneous closure or surgery in individual patients. To identify independent predictors of outcome and to quantitate the probability of spontaneous closure and surgery in patients with isolated VSD, we studied 156 consecutive infants who had a diagnosis of an isolated VSD between January 1, 1988, and December 31, 1990, and who were subsequently monitored for 28.5 +/- 15 months. Of the 149 patients with membranous (n = 100) and muscular (n = 49) defects who were studied, 46 (31%) patients had spontaneous closure, and an additional 37 (25%) patients underwent surgical repair. Univariate analysis identified defect cross sectional area indexed to body surface area, location in the muscular septum, presence of Down syndrome, and in membranous defects the presence of aneurysmal tissue as potential predictors of spontaneous closure or surgery. Multiple logistic regression analysis with these candidate variables identified indexed defect cross-sectional area as an independent predictor of spontaneous closure and surgery (p < 0.001). An inverse nonlinear relationship was seen between indexed VSD area and the probability of spontaneous closure (probability = (1 + e[-1.74 + 4.57CSA])-1 and a positive nonlinear relationship between indexed VSD area and the probability of surgery (probability = (1 + e[3.39 - 2.31CSA])-1). Muscular defects were more likely to close spontaneously than membranous defects (odds ratio 2.6, 95% CL = 1.01 - 6.8, p = 0.04).(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7484776 TI - Percutaneous use of stents to correct pulmonary artery stenosis in young children after cavopulmonary anastomosis. AB - Pulmonary artery distortion is a risk factor among candidates for the Fontan procedure. In 57 patients evaluated by catheterization after successful cavopulmonary anastomosis, 8 had proximal left pulmonary artery (LPA) stenosis, either discrete (4 patients) or long segment (4 patients). Median age was 27 months (range 19 to 60 months). Median weight was 11.4 kg (range 9.1 to 20.0). Mean diameter at LPA stenosis was 4.4 +/- 0.4. Proximal right pulmonary artery mean diameter was 10.4 +/- 1.0 mm. After angiographic and hemodynamic assessment, short 11F sheaths were placed in the right internal jugular (6 patients) or subclavian veins (2 patients). Pulmonary artery angioplasty and stent placement were performed. LPA stenoses were enlarged using 10 Palmaz stents dilated to 10 mm (7 patients) or to 12 mm (3 patients). Poststent angiograms showed that narrowest LPA dimensions were significantly enlarged to 9.9 mm +/- 1.0 mm, p < 001). There were no complications. Follow-up studies (catheterizations in 4 patients, echocardiograms in 8 patients) were performed 4 to 9 months after stent implantation. No restenosis was observed. Five patients had completion of their Fontan procedures; three patients are pending Fontan completion. This study demonstrates the efficacy and safety of the percutaneous use of Palmaz stents to correct pulmonary artery stenosis in young children after cavopulmonary anastomosis. PMID- 7484777 TI - Randomized double-blind, placebo-controlled trial of oral atenolol in patients with unexplained syncope and positive upright tilt table test results. AB - The objective of this investigation was the assessment of the response rate of oral atenolol in patients with vasovagal syncope after 1 month of treatment. We randomized into two groups all patients referred to our unit who had had at least one episode of syncope or two episodes of presyncope 1 month before presentation and had a positive isuprel Tilt Table Test (TTT). Group 1 (Gr 1) received oral atenolol, and group 2 (Gr 2) received placebo medication. After a 1-month period patients were reassessed for degree of their symptoms and underwent repeated TTT. Forty-two patients were enrolled in the study. Gr 1 and Gr 2 were comparable in age (38 +/- 13 years vs 43 +/- 14 years, p = 0.216 and sex (male/female = 6:15 vs 10:11, p = 0.204). The severity of attack was similar in both groups. Eight patients in Gr 1 and six patients in Gr 2 had mitral valve prolapse (p = 0.5). No significant differences were seen in systolic blood pressure (122 +/- 17 vs 117 +/- 16 mmHg, p = 0.334), diastolic blood pressure (70 +/- 11 vs 72 +/- 11 mm Hg, p = 0.677), and heart rate (79 +/- 12 vs 79 +/- 13, p = 0.98) between the two groups. The response rates (negative TTT) after 1 month of treatment were 62% versus 5% (p = 0.0004) in the atenolol and control group, respectively. Moreover, patients who received atenolol reported feeling better compared with those who received placebo (71% vs 29%, p = 0.02). In conclusion, atenolol significantly improved symptoms of patients with vasovagal syncope.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7484778 TI - Spontaneous echocardiographic contrast in the thoracic aorta: factors associated with its occurrence and its association with embolic events. AB - Spontaneous echocardiographic contrast is associated with embolic events when it occurs in the left atrium. Because little is known about spontaneous echocardiographic contrast in the aorta, we investigated this association retrospectively in 343 patients without aortic dissection or aneurysm who had undergone transesophageal echocardiography. Two independent readers concurred on the presence of spontaneous echocardiographic contrast in the aorta in 93% of the study patients, with the remainder agreed on by consensus. Spontaneous echocardiographic contrast was found in 65 patients (19%) and was associated with older age (p < 0.0001), male sex (p < 0.0001), slightly larger aortas (p < 0.0001), and complex aortic atherosclerosis (p = 0.0001). Thirty-four (28.6%) of 119 patients with clinical embolic events had spontaneous echocardiographic contrast in the aorta in contrast to 31 (13.8%) of 224 patients referred for other reasons (p = 0.0001). This finding remained significant when spontaneous echocardiographic contrast in the aorta was the only abnormality allowed (n = 207, p = 0.0065) or when other echocardiographic variables known to be related to embolic events were included in a multivariate analysis. Thus, spontaneous echocardiographic contrast in the aorta can often be detected by transesophageal echocardiography and is associated with a higher prevalence of embolic events. PMID- 7484779 TI - Colony-stimulating factors and coxackievirus B3 myocarditis in mice: macrophage colony-stimulating factor suppresses acute myocarditis with increasing interferon alpha. AB - The effects of macrophage colony-stimulating factor (M-CSF) and granulocyte colony-stimulating factor (G-CSF) on murine coxsackievirus B3 myocarditis were investigated. A total of 4 x 10(6) U/kg/day M-CSF and 20 micrograms/kg/day G-CSF were injected subcutaneously every day on day 0 to day 14 starting simultaneously with virus inoculation. Serum interferon-alpha was measured periodically. The survival rate of the M-CSF group was higher than that of the untreated control group (p < 0.05). On days 7 and 14 cardiac disease was significantly lower in the M-CSF group than in the untreated control group. Myocardial virus titers on day 7 in the M-CSF group were lower compared with those of the untreated control group. No significant difference was seen in the survival, cardiac disease, or myocardial virus titers between the G-CSF and the control groups. Monocyte counts on days 7 and 14 in the M-CSF group were increased compared with those in the control group. Serum interferon-alpha titers in the M-CSF and G-CSF groups on day 4 and those in the M-CSF group on day 7 were significantly increased in comparison with those of the untreated control group. We conclude that M-CSF but not G-CSF has the potency to limit myocardial virus titers and to reduce cardiac disease in the acute coxsackievirus B3 myocarditis. This ability is associated with an elevated interferon-alpha. Thus macrophages may play a defensive role in this model. PMID- 7484780 TI - Doppler echocardiographic characteristics of sinus of valsalva aneurysms. AB - This study evaluated the efficacy of a comprehensive transthoracic Doppler echocardiographic examination for correct detection of SVA and its associated lesions of VSD and aortic insufficiency. A consecutive series of 36 patients was obtained; all had surgical confirmation of the anatomic defects. The location of the SVA was correctly determined in 97% of patients, and the associated presence of a VSD was correctly determined in 79% of the studies. In the cases in which Doppler was available, all 11 patients with associated aortic insufficiency were detected. In this series 32 patients had right SVA, 3 had noncoronary sinus SVA, and 1 had a left SVA. Relative frequency of the rupture site location corresponded closely to that in previous necropsy studies. It is concluded that transthoracic Doppler echocardiography can correctly localize SVAs and most associated lesions. PMID- 7484781 TI - Hemodynamic and vascular effects of active and passive smoking. AB - Epidemiologic studies suggest that active and passive exposure to tobacco smoke is an important cause of cardiovascular morbidity and mortality. Numerous clinical studies have demonstrated that cigarette smoking causes coronary vasoconstriction, an increase in coronary vascular resistance, and a decrease in coronary blood flow, despite an increase in myocardial oxygen demand. Cigarette smoking also induces diffuse or segmental coronary artery spasm. In habitual smokers, smoking one cigarette increases heart rate, blood pressure, cardiac index, and myocardial oxygen demand and impairs cardiac performance, probably through adrenergic stimulation and catecholamine release. Several experimental studies, however, show that cigarette smoke inhalation causes pulmonary vasodilation because of inhalation of NO and CO in the vapor phase of cigarette smoke. Similar to active smoking, passive smoking has the same adverse effects on the cardiovascular system, with similar changes in hemodynamics and coronary vasomotor tone, platelet activation, impairment of endothelium-dependent vasodilation, and endothelial dysfunction. The adverse cardiovascular effects of smoking can be partially abolished by alpha- and beta-blockers or by calcium entry blockers. PMID- 7484782 TI - Right ventricular dysfunction after acute pulmonary embolism: pathophysiologic factors, detection, and therapeutic implications. AB - Acute PE may lead to right ventricular dilatation and failure. Through ventricular interdependence and decreased left ventricular filling, cardiac output and systemic circulation also may be compromised. The associated decrease in coronary perfusion pressure to the acutely overloaded right ventricle may produce ischemia and worsening right heart failure. This downward cycle of right ventricular failure and ischemia may ultimately progress to right ventricular infarction, circulatory arrest, and death. Certain clinical findings, hemodynamic values, and, particularly, echocardiographic signs can identify right ventricular dysfunction after PE. Detection of right ventricular hypokinesis helps to stratify patients' risk, because right ventricular dysfunction confers a worse prognosis than does normal right ventricular function after PE. The concept of "hemodynamic instability" after PE should be expanded to include right ventricular dilatation and wall motion abnormalities, even among normotensive patients. Aggressive intervention with thrombolytic therapy, vasoactive agents, or mechanical embolectomy may improve right ventricular function and clinical outcome. PMID- 7484783 TI - Development of coronary compensatory enlargement in vivo: sequential assessments with intravascular ultrasound. PMID- 7484784 TI - Perforation of chronic total occlusion with laser guide wire followed by multiple stent deployment: usefulness of three-dimensional intracoronary ultrasound guidance. PMID- 7484786 TI - Myocardial infarction during scuba diving: a case report and review. PMID- 7484785 TI - Stand-alone stenting of the left main coronary artery and 16-month patency despite sepsis and complicated hospital course. PMID- 7484790 TI - Reciprocating tachycardia in a patient with high-grade atrioventricular block. PMID- 7484787 TI - Detection of ascending aorta thrombi with transesophageal echocardiography in patients with systemic embolization. PMID- 7484791 TI - Response of left atrial systolic function to handgrip in normal subjects. PMID- 7484792 TI - Catheter fragmentation of massive pulmonary embolus: guidance with transesophageal echocardiography. PMID- 7484789 TI - Massive pericardiac hematoma with severe constrictive pathophysiologic complications after insertion of an epicardial pacemaker. PMID- 7484788 TI - Hypereosinophilic syndrome with rapid progression of cardiac involvement and early echocardiographic abnormalities. PMID- 7484793 TI - Pericardial effusion as the initial feature of Q fever. PMID- 7484795 TI - Giant Cell Myocarditis Study Group. PMID- 7484796 TI - Transesophageal echocardiography microbubbles with prosthetic valves. PMID- 7484794 TI - Variant angina after successful coronary atherectomy. PMID- 7484797 TI - Thrombolytic drugs and acute aortic dissection. PMID- 7484798 TI - Familial dilated cardiomyopathy. PMID- 7484801 TI - Negative U waves in coronary disease. PMID- 7484799 TI - Thyrotoxicosis and dilated cardiomyopathy. PMID- 7484800 TI - Mitral valve prolapse. PMID- 7484802 TI - Alagille syndrome (arteriohepatic dysplasia) PMID- 7484803 TI - Carotid sinus hypersensitivity. PMID- 7484804 TI - Vascular overload concept. PMID- 7484805 TI - Amiodarone and sexual dysfunction. PMID- 7484809 TI - Does counseling improve compliance? PMID- 7484808 TI - Computer-generated prescription orders. PMID- 7484806 TI - Lovastatin and peripheral neuropathy. PMID- 7484807 TI - Is it safe to prescribe digoxin after acute myocardial infarction? Update on continued controversy. PMID- 7484810 TI - Rainforest holds many lessons for pharmacy. PMID- 7484811 TI - Winning payment for cognitive services: what works. PMID- 7484812 TI - Eight steps toward payment for cognitive services. PMID- 7484813 TI - Standardizing documentation for filing pharmaceutical care claims. PMID- 7484815 TI - Evaluating employee benefits: insurance and flexible spending. PMID- 7484816 TI - Response to: 'a mother's perspective'. PMID- 7484814 TI - Practical considerations in anticonvulsant therapy--Part 2. AB - Epilepsy is, for many patients, a lifelong condition that requires treatment with powerful drugs whose doses must be carefully titrated to avoid both breakthrough seizures and toxicity. The medication regimens used to treat epilepsy are further complicated by the fact that most seizure medications are metabolized in the liver and have the potential for serious pharmacokinetic drug-drug interactions with many other medications. Successful management of epilepsy requires a high degree of cooperation among the patient, the pharmacist, and the treating physician. Such cooperation can ensure that the appropriate treatment and drug preparation are selected, compliance is maintained, and dangerous drug-drug interactions are avoided. PMID- 7484818 TI - The pharmacist's role on the hospital patient management team. PMID- 7484817 TI - Interactions and drug-metabolizing enzymes. PMID- 7484819 TI - What's in the pipeline? PMID- 7484820 TI - Physical working capacity after acute myocardial infarction in patients with low ejection fraction and effect of captopril. AB - Previous studies after acute myocardial infarction (AMI) have reported conflicting results on the effects of angiotensin-converting enzyme inhibition on physical working capacity. In an effort to provide more insight into this subject, we examined the effects of captopril on working capacity of patients who had low ejection fractions but no congestive heart failure after AMI. One hundred sixty-six participants were recruited from 5 centers after randomization to either captopril or placebo for the Survival and Ventricular Enlargement study. Upright cycle ergometer tests were performed with continuous measurements of respiratory gases at 4, 12, and 24 months after AMI. Our study concurs with 2 of 3 previous post-AMI studies and supports the conclusion that working capacity is not affected by angiotensin-converting enzyme inhibition at 4 or 12 months after AMI in patients without congestive heart failure. In addition, no significant effect of captopril was noted at 24 months after AMI. Peak oxygen uptake tended to decrease between 12 and 24 months in the placebo group by an average (+/- SD) of -22 +/- 322 ml/min (n = 66), but to increase in the captopril group (+62 +/- 289, n = 57), a difference that was significant (Mann-Whitney chi-square, p = 0.02). This post-hoc observation suggests that a late beneficial effect may have been masked by inadequate study duration. Known benefits of captopril appear not to include an increase in working capacity within the first 24 months after AMI. PMID- 7484822 TI - The aminoterminal propeptide of type III procollagen provides new information on prognosis after acute myocardial infarction. AB - The aim of the study was to examine sequential changes in serum levels of the aminoterminal propeptide of type III procollagen (S-PIIINP) after acute myocardial infarction (AMI), and to assess the value of S-PIIINP as a predictor of outcome. The study group comprised 74 patients with AMI, and 24 patients in whom AMI was suspected but disproved. S-PIIINP changed characteristically after AMI, and in patients not receiving thrombolytic therapy or having cardiogenic shock, the changes correlated to peak enzyme values (r = 0.4, p < or = 0.03). S PIIINP was higher at days 0 to 2 in nonsurviving AMI patients than in survivors (p < 0.05). With use of either the upper quartile for S-PIIINP at day 0 for nonsurviving AMI patients or the mean value of S-PIIINP in a normal population plus 2 SDs as a cutoff, the predictive value of a negative test ranged from 0.79 to 0.87 at days 0 to 2, and the predictive value of a positive test ranged from 0.39 to 0.67. Thus, S-PIIINP on admission and for the following few days after AMI is higher in patients with poor outcome. PMID- 7484821 TI - Ability of the no-reflow phenomenon during an acute myocardial infarction to predict left ventricular dysfunction at one-month follow-up. AB - Despite angiographically successful opening of an infarct-related vessel within a 6-hour time frame, some patients do not recover left ventricular regional wall function in the infarct zone after an acute myocardial infarction (AMI). Recent evidence suggests that this finding is due to the no-reflow phenomenon, or failure to recover tissue perfusion despite patient epicardial arteries. We performed myocardial contrast echocardiography to assess tissue perfusion before and after opening of an infarct-related artery. Coronary angiograms, regional wall motion scoring, and myocardial contrast enhancement were graded by 3 observers. Of 24 patients with AMI, 7 (29%) failed to recover tissue perfusion in > or = 1 region of myocardium. Of 106 regions subtended by the infarct-related artery, 16 (15%), 43 (41%), and 47 (44%) regions had no-reflow, partial, or normal flow, respectively, after arterial patency was established. There was a spectrum of reperfusion patterns ranging from no-reflow to normal perfusion. One month follow-up angiographic and myocardial contrast echocardiographic studies were performed in 12 of the 24 patients. At 1 month, all segments of myocardium that had immediate normal perfusion had regained normal wall motion. In contrast, 17 segments that had partial or no-reflow were identified. Of these 17, 3 regained normal function, 10 segments were hypokinetic, and 4 segments were akinetic. We conclude that myocardial contrast echocardiography can be used to identify the no-reflow phenomenon in up to 29% of patients with AMI. Additionally, we found that the immediate-reflow pattern can predict degree of left ventricular dysfunction at 1-month follow-up. PMID- 7484823 TI - Adaptation mechanisms during myocardial ischemia in chronic unstable angina pectoris. AB - Unstable angina with a clinical duration of < 2 months is characterized angiographically by a high incidence of complex lesions. Some patients have ischemic rest pain syndromes of longer duration. Thus, we retrospectively analyzed, in blinded fashion, the clinical and angiographic findings in 52 patients with unstable angina of < 2 months' duration (group A), and compared the results with those of 32 patients with unstable angina of > 6 months' duration (group B). Group B had a greater number of diseased vessels and better collateral circulation, but had fewer eccentric lesions. There were no differences in age, left ventricular function, or history of prior myocardial infarction. Thus, chronic unstable angina is associated with more extensive coronary disease than unstable angina of shorter duration. The role of different anatomic substrata and collateral circulation is discussed. PMID- 7484824 TI - Dobutamine echocardiography in predicting improvement in global left ventricular systolic function after coronary bypass or angioplasty in patients with healed myocardial infarcts. AB - The aim of this study was to determine whether low-dose dobutamine echocardiography (DE) could predict quantitative improvement in global left ventricular (LV) systolic function after coronary revascularization. Low-dose DE was performed in 71 consecutive patients with coronary artery disease and LV dysfunction. Successful coronary bypass surgery or angioplasty was performed in 44 patients, 37 of whom had a resting echocardiogram 1 to 3 months afterward. Group A consisted of 20 patients with contractile reserve during DE, and group B consisted of 17 patients without contractile reserve. As expected, regional wall motion score index (mean +/- SD) improved in group A (1.62 +/- 0.39 to 1.38 +/- 0.31, p < 0.01) but not group B (1.56 +/- 0.42 to 1.57 +/- 0.41, p = NS). In addition, LV ejection fraction (LVEF) improved after bypass surgery or angioplasty in group A (38 +/- 5% to 42 +/- 5%, p < 0.01), but not in group B (38 +/- 7% to 39 +/- 8%, p = NS). In group A, a significant linear correlation was observed between the number of segments with contractile reserve and the improvement in LVEF (r = 0.91, p < 0.0001). A good correlation also existed between the improvement in regional wall motion score index during dobutamine infusion and the improvement in LVEF after bypass surgery or angioplasty (r = 0.90, p < 0.0001). In conclusion, low-dose DE can be used to predict quantitative improvement in global LV systolic function after coronary bypass or angioplasty. PMID- 7484825 TI - Normal stroke volume and cardiac output response during dobutamine stress echocardiography in subjects without left ventricular wall motion abnormalities. AB - Dobutamine stress echocardiography has become widely utilized for evaluation of coronary artery disease, but the expected responses of stroke volume and cardiac output to the high doses of dobutamine administered in these studies are not known. To determine these responses, stroke volume and cardiac output were measured with 2-dimensional Doppler echocardiography at each stage of dobutamine stress echocardiography and after administration of atropine in 47 patients without resting or inducible wall motion abnormalities. Heart rate increased significantly at each stage of dobutamine infusion and after atropine. Mean blood pressure decreased at the 5 micrograms/kg/min dose, then showed little change. Stroke volume increased 27 +/- 18% from baseline, with significant increases occurring at both the 5 and 10 micrograms/kg/min doses (p < 0.00001). With higher doses of dobutamine, stroke volume tended to plateau or decrease. Mean changes in stroke volume were not significant between the doses of 10, 20, and 30 micrograms/kg/min. The mean change in stroke volume from the 30 to the 40 micrograms/kg/min dose was a significant decrease of 6.3% (p = 0.004); the decrease from the 40 micrograms/kg/min dose to atropine approached statistical significance (p = 0.06). Cardiac output increased throughout dobutamine infusion. Stroke volume during dobutamine stress echocardiography is commonly maximum at a dose of 20 micrograms/kg/min and tends to decline at higher infusion rates. At higher doses, increases in cardiac output are mediated primarily by increases in heart rate. PMID- 7484828 TI - A new system for classification of cardiac death as arrhythmic, ischemic, or due to myocardial pump failure. AB - Existing classifications of cardiac death fail to incorporate current understanding of the pathophysiology of sudden cardiac death. We developed a new scheme for classifying cardiac death that defines 3 categories of underlying mechanism: primary arrhythmia, acute myocardial ischemia/infarction, and myocardial pump failure. Using this new system, we classified the mechanism of 106 definite cardiac deaths from the Recurrent Coronary Prevention Project. Fifty deaths (47%) were classified as arrhythmic, 46 (43%) as ischemic, and 9 (8%) as due to myocardial pump failure (1 death was not classifiable). All 36 witnessed arrhythmic deaths were sudden and 8 of 9 witnessed myocardial pump failure deaths were nonsudden. The 38 witnessed ischemic deaths were split evenly between sudden and nonsudden. Interrater agreement for the classification of mechanism was 100%. This classification scheme, if validated in subsequent studies, will provide a useful algorithm for classifying deaths by underlying mechanism. PMID- 7484827 TI - Diagnostic usefulness of postexercise systolic blood pressure response for detection of coronary artery disease in patients with electrocardiographic left ventricular hypertrophy. AB - Patients with left ventricular (LV) hypertrophy often have a positive result on exercise testing despite a normal coronary arteriogram. This indicates that exercise-induced ST depression is not always an accurate indicator of the presence of coronary artery disease (CAD) in such patients. We evaluated the usefulness of the postexercise systolic blood pressure (BP) response for detection of CAD in 51 patients with both electrocardiographic evidence of LV hypertrophy and positive ST depression on treadmill exercise testing. Coronary cineangiograms showed normal coronary arteries in 23 patients (45%) (group 1) and significant CAD in 28 patients (55%) (group 2). The systolic BP ratio (systolic BP at 3 minutes of recovery divided by systolic BP at peak exercise) was significantly higher in group 2 than in group 1 (1.01 +/- 0.19 vs 0.80 +/- 0.09; p < 0.001). Analysis of the relative cumulative frequency revealed that a systolic BP ratio of 0.86 was the cutoff point for distinguishing a patient with CAD from one with normal coronary arteries. The sensitivity, specificity, and accuracy of a systolic BP ratio > or = 0.86 for detection of CAD in patients with LV hypertrophy were 79%, 83%, and 82%, respectively. Our results suggest that the use of an abnormal BP ratio, in combination with ST depression, improves the accuracy of treadmill exercise testing for detecting CAD in patients with electrocardiographic evidence of LV hypertrophy. PMID- 7484829 TI - Design, rational, and baseline characteristics of the Prospective Pravastatin Pooling (PPP) project--a combined analysis of three large-scale randomized trials: Long-term Intervention with Pravastatin in Ischemic Disease (LIPID), Cholesterol and Recurrent Events (CARE), and West of Scotland Coronary Prevention Study (WOSCOPS). AB - The Prospective Pravastatin Pooling (PPP) project is a pooled evaluation of 3 large, placebo-controlled, randomized trials of cholesterol-lowering treatment with pravastatin. It is designed to more reliably evaluate the effect of treatment on coronary and all-cause mortality and on total coronary artery disease (CAD) events for specific populations of interest, including women and the elderly. The trials--Long-Term Intervention With Pravastatin in Ischemic Disease trial, the Cholesterol and Recurrent Events trial, and the West of Scotland Coronary Prevention Study--each have common design features, including drug, dose, and duration. The project prospectively defines the objectives, end points, and analytic plans in a protocol developed before results are known of any individual trial. More than 2,000 (or 10%) of the participants in the pooled data set are women, 1,841 are aged > or = 70 years at trial entry, and > 6,000 have a total cholesterol < 5.5 mmol/L (213 mg/dl). The mean low-density lipoprotein cholesterol level is 4.2 mmol/L (162 mg/dl). The mean blood pressure level is 134/81 mm Hg and 20% are current smokers. Half of the PPP participants have had a prior myocardial infarction. More than 7% have a history of diabetes and 26% have a history of hypertension. PPP is projected to have data on about 1,100 CAD deaths, 500 non-CAD deaths, and > 1,000 cancers by study completion.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7484826 TI - Prognostic value of dobutamine stress echocardiography in patients referred because of suspected coronary artery disease. AB - To determine whether dobutamine stress echocardiography (DSE) provides prognostic information beyond that available from routine clinical data, we reviewed the outcome of 210 consecutive patients referred for DSE to evaluate chest pain, perioperative risk, and myocardial viability. Dobutamine was infused in increments of 10 micrograms/kg/min in 5-minute stages to a maximum of 40 micrograms/kg/min. The dobutamine stress echocardiogram was considered abnormal only if dobutamine induced a new wall motion abnormality as determined by review of the digitized echocardiographic images in a quad screen format and on videotape. Thirty percent of tests were abnormal. An abnormal test was more common (p < or = 0.02) in men and patients with angina pectoris, in patients taking nitrate therapy, or those with prior myocardial infarction or abnormal left ventricular wall motion at rest. Twenty-two deaths, 17 of which were cardiac, occurred over a median follow-up of 240 days (range 30 to 760). Sixteen cardiac deaths occurred in the 63 patients with versus 1 cardiac death among the 147 without a new wall motion abnormality (p < or = 0.0001). Other variables associated with cardiac death (p < or = 0.05) were age > 65 years, nitrate therapy, ventricular ectopy during DSE, suspected angina pectoris, and hospitalization at the time of DSE. When cardiac death, myocardial infarction, and revascularization procedures were all considered as adverse outcomes, a new wall motion abnormality continued to be the most powerful predictor of an adverse cardiac event.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7484830 TI - Age, race, and sex differences in autonomic cardiac function measured by spectral analysis of heart rate variability--the ARIC study. Atherosclerosis Risk in Communities. AB - To investigate the distribution of heart rate variability (HRV) spectral power in an unselected sample of the population, and to ascertain the population correlates of HRV, we examined 1,984 healthy persons, aged 45 to 64 years, randomly selected from the Atherosclerosis Risk in Communities (ARIC) study cohort. Resting, supine, 2-minute, beat-to-beat heart rate data were collected between 7 A.M. and 12 noon. The race- and sex-adjusted geometric means of low frequency component (LF, 0.025 to 0.15 Hz) were 4.00 and 3.13 (beats/min)2; of high-frequency component (HF, 0.16 to 0.35 Hz), 1.65 and 1.21 (beats/min)2; and of the HF/LF ratio, 0.41 and 0.39, for 45-to-54 and 55-to-64 years age groups, respectively (test of mean difference by age, p < 0.01, p < 0.01, and p = 0.11 for LF, HF, and HF/LF ratio, respectively). Comparing black with white examinees, the age- and sex-adjusted geometric means of LF were 3.06 and 3.70 (beats/min)2; of HF, 1.66 and 1.36 (beats/min)2; of HF/LF, 0.54 and 0.37, respectively (test of mean difference by race, p < 0.01, p < 0.01, and p < 0.01). The age- and race adjusted geometric means of LF for women and men were 3.12 and 4.10 (beats/min)2; of HF, 1.46 and 1.38 (beats/min)2; and of HF/LF, 0.47 and 0.34, respectively (test of mean difference, p < 0.01, p = 0.34, and p < 0.01). We conclude that HRV spectral indexes are associated with age, race, and sex. With increasing age, the parasympathetic and sympathetic spectral power components decrease.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7484831 TI - Clinical efficacy and safety of atrial defibrillation using biphasic shocks and current nonthoracotomy endocardial lead configurations. AB - We undertook a prospective randomized clinical trial evaluating efficacy and safety of internal atrial defibrillation in patients with drug-refractory atrial fibrillation (AF). Consecutive patients with paroxysmal or chronic AF were randomly tested with 3 internal atrial defibrillation lead configurations and biphasic shocks. Patients with implanted cardiac pacemakers were tested with the right atrium (RA) and left pulmonary artery or coronary sinus (CS) configuration. Shocks were initially delivered without anesthesia to assess patient tolerance. The need for backup ventricular defibrillation and pacing support was evaluated. Eighteen patients with (n = 15) or without (n = 3) structural heart disease, mean left ventricular ejection fraction 36 +/- 14%, and mean left atrial diameter 4.5 +/- 0.6 cm were studied. The mean defibrillation threshold in the best randomized lead configuration was 9.9 +/- 7.7 J. Mean defibrillation threshold for the right ventricle (RV) and superior vena cava configuration was 13.3 +/- 5 J, which was significantly lower than the RA and axilla configuration (20.1 +/- 7.4 J, p < 0.04) but not the RV to RA configuration (16.5 +/- 11 J, p > 0.2). The mean defibrillation threshold using the RA-left pulmonary artery/CS configuration was 8.9 +/- 9 J (p > 0.2 vs RV-superior vena cava). There was a bimodal distribution of defibrillation thresholds. Low atrial defibrillation thresholds correlated with absence of heart disease, higher ejection fraction, and smaller left ventricular end-diastolic diameter. Shocks were hemodynamically well tolerated, but 2 of 18 patients (11%) had nonsustained ventricular tachycardia after shock delivery. Six of 18 patients (33%) had postshock bradyarrhythmias. Fourteen of 16 patients perceived shocks > or = 3 J as intolerable.(ABSTRACT TRUNCATED AT 250 WORDS) [corrected] PMID- 7484832 TI - Similar orthostatic defense in active, healthy young adult and late middle-aged men. AB - Orthostatic defense is commonly validated with a 60 degrees to 80 degrees head-up tilt test, addressing the step response rather than the response to permanent orthostatis. During the initial phase of tilt, neural factors predominate, while later, the slower humoral factors fade in. It has been demonstrated that, during adaptation of the circulatory system to the standing conditions, overshoot and undershoot occur. These oscillations hamper straight-forward interpretation of a tilt test, and may contribute to the inconclusiveness of current studies regarding the aging of orthostatic defense. Gradual, progressive, orthostatic load testing seems a valuable alternative. We used a novel, incremental, head-up tilt protocol (0 degrees to 80 degrees, 13 increments) to impose graded orthostatic stress on 46 healthy young adult men (mean age +/- SD 25 +/- 3 years), and on 16 healthy late middle-aged men (60 +/- 4 years), while recording the electrocardiogram and the blood pressure. A first-order estimate of the heart rate range associated with the sympathovagal transition was made by combined analysis of heart rate and heart rate variability trends. We observed similar responses in heart rate, heart rate variability, and blood pressure.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7484833 TI - Correlation between lipoprotein(a) and aortic valve sclerosis assessed by echocardiography (the JMS Cardiac Echo and Cohort Study). AB - An elevated serum level of lipoprotein(a) (Lp[a]) may be an independent risk factor for atherosclerotic disease, but the relation of Lp(a) to aortic valve (AV) sclerosis has not been determined. We measured serum concentrations of Lp(a) and investigated their relation to the presence of echocardiographic AV sclerosis in residents of a rural village in Japan. We measured serum Lp(a) levels in 347 men and 437 women aged 35 to 90 years (mean +/- SD: 62 +/- 11 years) who participated in mass screening examinations in Wara village, Gifu, Japan. AV sclerosis was assessed by long- and short-axis 2-dimensional echocardiographic views and continuous-wave Doppler echocardiography. AV sclerosis was graded as follows: 0 = normal AV; 1 = increased echo density; 2 = thickening or calcific deposits > or = 3 mm; and 3 = same as 2 with mildly restricted motion (pressure gradient < 16 mm Hg). Lp(a) levels ranged from < 1 mg/dl to 153 mg/dl. The 25th, 50th, and 75th percentile values were 7, 16, and 28 mg/dl, respectively. Lp(a) levels were significantly higher in women than in men (p < 0.01), and did not increase significantly with age. The prevalence of AV sclerosis (grades 2 and 3) increased significantly with age (p < 0.001). AV sclerosis was present in 65 (36.1%) of 180 subjects with Lp(a) levels > or = 30 mg/dl and in 77 (12.7%) of 604 subjects with Lp(a) levels < 30 mg/dl (p < 0.001). There were no significant differences in the prevalence of AV sclerosis in terms of sex, blood pressure, or levels of total cholesterol, high-density lipoprotein cholesterol, triglycerides, or blood sugar. We conclude that increased serum levels of Lp(a), as well as aging, are closely related to AV sclerosis. PMID- 7484835 TI - Racial differences in the prevalence of left ventricular hypertrophy among chronic cocaine abusers. AB - There are racial differences in the prevalence and pattern of left ventricular (LV) hypertrophy in hypertension. This study was performed to determine whether racial differences also exist in LV hypertrophy among chronic cocaine users. We studied 112 chronic cocaine abusers < 45 years old in whom normal blood pressures (< or = 140/90 mm Hg) were recorded 3 times daily for 3 weeks. LV wall thickness and mass were measured echocardiographically. Technically adequate studies were obtained in 79 blacks and 33 whites. Self-reported cocaine use was higher in whites than in blacks (688 +/- 516 vs 431 +/- 468 $/week, p = 0.03). There were no group differences in terms of duration of cocaine use, age, height, weight, blood pressure, LV dimensions, or left atrial size. However, posterior wall thickness (1.13 +/- 0.17 vs 1.03 +/- 0.14 cm, p = 0.0035) and LV mass index (113 +/- 25 vs 94 +/- 19 g/m2, p = 0.0001) were significantly greater in blacks. LV hypertrophy, defined as an M-mode LV mass index > or = 134 g/m2, was present in 24 blacks (30%) and 2 whites (6%) (p = 0.011). When defined as a posterior wall thickness > or = 1.2 cm and a 2-dimensional echocardiographic LV mass index > or = 105 g/m2, LV hypertrophy was present in 37 of 79 blacks (47%) and in 6 of 33 whites (18%) (p = 0.0086). Cocaine-related LV hypertrophy is more prevalent in black men than in white men.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7484834 TI - Incidence of infective endocarditis in the Delaware Valley, 1988-1990. AB - This population-based study aimed to determine the incidence of native, prosthetic, and bioprosthetic valve nosocomial infective endocarditis (IE), and IE associated with the use of injected drugs. Patients with IE during 27 months over the years 1988 to 1990, and residing in any of 6 counties in the Philadelphia metropolitan area were identified. An expert panel reviewed all patients to verify the diagnosis. Incidence rates were estimated after adjustment for failure to recruit and underreporting. Of 853 potential patients, 670 (79%) met the inclusion criteria. The overall incidence rate of IE was 11.6 cases/100,000 person-years (95% confidence interval [CI] 10.8 to 12.4). The rates for specific types of IE were: 4.45 (95% CI 3.97 to 4.94) for community-acquired native valve, 0.94 (95% CI 0.72 to 1.12) for prosthetic valve, 0.94 (95% CI 0.71 to 1.16) for nosocomial, and 5.34 (95% CI 4.80 to 5.87) for IE associated with use of injected drugs. Previous population studies found overall incidence rates of 1.7 to 4 cases/100,000 person-years, similar to our rate for community acquired native valve IE. Type-specific rates have not been previously reported. The higher overall rate in this study is partly related to the high prevalence of injection drug use in our area. PMID- 7484836 TI - Relation of right ventricular morphology and function in pectus excavatum to the severity of the chest wall deformity. AB - Although pectus excavatum (PE) is thought to impair right ventricular (RV) performance, the degree of RV dysfunction, if any, produced by this chest wall deformity remains controversial. To address this issue, we performed 2 dimensional echocardiography and chest wall radiography in 28 subjects with mild to-severe degrees of PE to assess RV morphology and function in relation to the degree of the chest wall deformity. Measurements of RV anatomy and function obtained in these patients were compared to those of 24 normal control subjects of similar age and sex. In subjects with PE, mean RV outflow tract diameter at the aortic root level was narrower (1.4 +/- 0.3 cm/m2) and end-diastolic (10 +/- 2.3 cm2/m2) and end-systolic (5.8 +/- 1.4 cm2/m2) areas were larger than those in normal controls (1.6 +/- 0.3, 8.6 +/- 1.7, and 4.5 +/- 1.2 cm2/m2, respectively; p < 0.013). The magnitude of these abnormalities was related to the degree of the chest wall deformity evaluated on the chest radiogram (r = 0.54, 0.51, and 0.49, respectively). RV planar emptying fraction, an index of RV systolic function, was reduced in subjects with PE (42 +/- 10%) compared to the normal controls (48 +/- 10%; p = 0.047). No relation could be found, however, between this index and the severity of the chest wall deformity.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7484837 TI - Circadian rhythm and variability of heart rate in Duchenne-type progressive muscular dystrophy. AB - Using 24-hour Holter monitoring and time domain and power spectral measurements, we evaluated the variability of the heart rate and its circadian rhythm in 55 male patients with Duchenne-type progressive muscular dystrophy (DMD) to characterize their autonomic function versus findings in 20 normal controls. Comparisons were also made in patients with mild, moderate, and severe stages of DMD. The percent difference between successive RR intervals that exceeded 50 ms, a measure of parasympathetic tone, was significantly lower even in patients with early stage of DMD than in controls (p < 0.01). This trend became marked with disease progression. Power in the high-frequency (HF) range (0.15 to 0.40 Hz), a measure of parasympathetic tone, was lower (p < 0.01), and the ratio of the power in the low-frequency (LF) range (0.04 to 0.15 Hz) and that of HF range (LF/HF ratio), a measure of sympathetic tone, was higher in DMD patients versus controls (p < 0.01). This trend was also marked with disease progression. Patients with mild or moderate disease had a slight circadian alteration in HF and LF/HF ratio. Patients with severe disease had virtually no circadian rhythm in HF. Their LF/HF ratio was higher at night (p < 0.01), lower in the morning (p < 0.01), and still lower during the day (p < 0.01), the opposite of control findings. The autonomic abnormalities in DMD were thus characterized by a significant increase in sympathetic activity and a significant decrease in parasympathetic activity. Thus, heart rate variability and circadian rhythm were useful in assessing autonomic dysfunction in DMD. PMID- 7484838 TI - Impact of echocardiographic left ventricular mass on mechanistic implications of exercise testing parameters. AB - Electrocardiographic left ventricular (LV) hypertrophy has long been known to be associated with an abnormal ST-segment response to exercise; this association has been considered to represent a false-positive finding. There is a paucity of data relating echocardiographic LV mass to exercise ST-segment responses and other exercise parameters. As part of a routine evaluation, 1,408 men and 1,618 women from the Framingham Heart Study who were free of clinical cardiac disease underwent echocardiography and exercise treadmill testing according to the Bruce protocol at the same clinic visit. Abnormal ST-segment responses were defined both by standard criteria and the delta ST/heart rate index. LV mass was calculated from M-mode echocardiography. Echocardiographic LV hypertrophy was associated with an abnormal delta ST/heart rate index (in men, odds ratio [OR] 1.78, 95% confidence interval [CI] 1.05 to 3.01, p = 0.03; in women, OR 2.13, 95% CI 1.31 to 3.44, p = 0.002) but not with an abnormal response according to standard criteria. Echocardiographic LV hypertrophy was also associated with a lower likelihood of achieving an age-predicted target heart rate (in men, OR 0.45, 95% CI 0.31 to 0.65, p < 0.001; in women, OR 0.53, 95% CI 0.37 to 0.76, p < 0.001) and with a lower exercise capacity. Despite these associations, echocardiographic LV hypertrophy was associated with a higher peak heart rate systolic blood pressure double product. In conclusion, echocardiographic LV hypertrophy is associated with an abnormal ST-segment response, a lower likelihood of achieving target heart rate, decreased exercise capacity, and an increased double product, which is a reflection of myocardial oxygen demand. PMID- 7484839 TI - Regulation of constitutive nitric oxide synthase activity by the human heart. AB - We and others have provided indirect evidence for the presence of a constitutive nitric oxide synthase (cNOS) in the mammalian heart. We now provide more direct evidence for the regulation of a myocardial cNOS in the hearts of patients undergoing elective cardiopulmonary bypass (CPB). cNOS enzyme activity was demonstrable in both cytosolic (8.3 +/- 0.02 pmol/min/mg) and membrane (11.1 +/- 0.4 pmol/min/mg) preparations derived from human atrial pectinate muscles obtained at the time of CPB (n = 6). Plasma nitrite (NO2-) + plasma nitrate (NO3 ) levels from the beating hearts of patients before bypass were reduced from 146 +/- 33 to 5.1 +/- 50 pmol/min/g after cardiac arrest during CPB (n = 23; p < 0.002 by Student's t test). Thus, the human myocardium constitutively produces nitric oxide that is regulated by the contractile state of the heart. PMID- 7484840 TI - Role of telemetry monitoring in the non-intensive care unit. AB - To determine the outcomes of patients admitted to a non-intensive care telemetry unit and to assess the role of telemetry for guiding patient management decisions, data from 2,240 patients admitted to a telemetry unit were collected prospectively during 7 months. Physicians recorded the outcomes (intensive care unit transfer and mortality) and assessed whether telemetry assisted in guiding patient management. Indications for admission to the telemetry unit included chest pain syndromes (55%), arrhythmias (14%), heart failure (12%), and syncope (10%). Telemetry led to direct modifications in management in 156 patients (7%; 95% confidence interval [CI] 5.9% to 8%). Telemetry was perceived as useful but did not alter management for 127 patients (5.7%; 95% CI 4.7% to 6.6%). Two hundred forty-one patients were transferred to an intensive care unit from the telemetry unit (10.8%; 95% CI 9.5% to 12%). Nineteen patients (0.8% of all admissions; 95% CI 0.5% to 1.2%) were transferred because of an arrhythmia identified by telemetry. Routine transfer after cardiac revascularization or surgery accounted for 134 transfers; clinical deterioration accounted for 88 transfers. There were 20 deaths in the unit (0.9%; 95% CI 0.5% to 1.3%): 4 of the 20 deaths occurred while patients were being monitored. The role of telemetry in guiding patient management may be overestimated by physicians, since it detected significant arrhythmias that led to change in medications or urgent interventions in a small fraction of patients. PMID- 7484841 TI - Elective percutaneous transluminal coronary angioplasty after acute myocardial infarction: is later better? PMID- 7484842 TI - Clinical markers, management, and long-term follow-up of early ischemia after coronary artery bypass grafting. AB - In summary, the occurrence of angina or myocardial infarction within 1 year after coronary bypass is associated with a high incidence of significant angiographic abnormalities. Early angiography is necessary to identify high-risk patients who could undergo revascularization. Patients with other markers of ischemia should have a noninvasive approach (stress imaging test) as initial evaluation, before coronary angiography is considered. When technically feasible, coronary angioplasty can be performed safely and with a high success rate. Repeat coronary bypass in this group of patients is associated with higher in-hospital complications. Patients with less compromised coronary anatomy can be treated medically with a good long-term outcome. PMID- 7484844 TI - Association of increased left ventricular mass in the absence of electrocardiographic left ventricular hypertrophy with ST depression during exercise. PMID- 7484843 TI - Prognostic value of cardiac troponin T in unstable angina pectoris. PMID- 7484845 TI - Effects of dietary fish oil on ventricular premature complexes. AB - For ethical and practical reasons, in this study the antiarrhythmic potential of fish oil was evaluated in patients free from complex ventricular arrhythmias and severe heart failure. Although subjects without overt structural heart disease had ventricular arrhythmias that were not associated with an increased risk for sudden cardiac or coronary death, recent data suggest that frequent VPCs in patients similar to our study population may reflect subclinical cardiac disease amenable to the multiple beneficial actions of n-3 fatty acids. The potential and safety of fish oil as a treatment for more complex cardiac arrhythmias or arrhythmias in higher risk patients with more severe heart disease deserve further study. PMID- 7484846 TI - Comparison of hemodynamic responses to cycling and resistance exercise in congestive heart failure secondary to ischemic cardiomyopathy. PMID- 7484847 TI - Optimal degree of pulmonary artery banding--adequate circumference ratio to calculated size from normal pulmonary valve dimensions. AB - These findings suggest that PA banding may be suitable in children with congenital heart disease and excessive pulmonary flow, and that best results are obtained when the band circumference is < 90% of the standard pulmonary valve ring circumference, as calculated from an equation derived from normal pulmonary valve dimensions. This guideline applies equally well to small infants weighing < 3 kg and to larger patients. PMID- 7484848 TI - Morphologic features of the interrupted aortic arch in Chinese patients. PMID- 7484849 TI - Noninvasive quantitation of left ventricular wall thickening using cine magnetic resonance imaging with myocardial tagging. PMID- 7484850 TI - Percutaneous transcatheter repair of a coronary aneurysm using a composite autologous cephalic vein-coated Palmaz-Schatz biliary stent. AB - We describe a novel percutaneous transcatheter technique using an autologous vein graft-coated Palmaz-Schatz stent for the treatment of a coronary aneurysm. This technique avoids the procedural risks that are currently associated with surgical repair with revascularization or spring coil embolization in the treatment of coronary aneurysm. PMID- 7484851 TI - Value of transthoracic two-dimensional echocardiography in predicting viability in patients with healed Q-wave anterior wall myocardial infarction. AB - The role of transthoracic echocardiography as a predictor of recovery after revascularization has not yet been established. Two-dimensional echocardiography was performed in 15 patients with a healed anterior wall myocardial infarction and severe, isolated stenosis of the left anterior descending coronary artery before, and 3 to 6 months after angiographically confirmed successful revascularization. The asynergic segments were classified into 2 groups according to 2 different echocardiographic patterns: those showing a normal acoustic reflectance with normal end-diastolic thickness (pattern A segments) and those showing an increase in acoustic reflectance and reduced end-diastolic thickness (pattern B segments). We hypothesized that pattern A segments were more likely to recover (viable myocardium) and that pattern B segments were consistent with irreversibility. A total of 240 segments in the 15 patients were evaluated before and after revascularization. Sixty-seven segments were asynergic; of these, 52 were judged to have pattern A and 15 pattern B. Of the 52 pattern A segments, 27 were hypokinetic and 25 akinetic. All of the pattern B segments were akinetic (n = 9) or dyskinetic (n = 6). Pattern A was predictive of postoperative recovery in 39 of 52 segments (75%) (p < 0.0001); pattern B was predictive of irreversibly damaged tissue in 13 of 15 segments (87%) (p < 0.0001). Thus, in patients with healed anterior wall myocardial infarction, resting transthoracic echocardiography is a simple and reliable predictor of the behavior of asynergic segments after revascularization. PMID- 7484852 TI - Frequency, significance, and cost of recurrent ischemia after thrombolytic therapy for acute myocardial infarction. TAMI Study Group. AB - Early postinfarction angina implies an unfavorable prognosis. Most published information on this outcome represents data collected in the prethrombolytic era, in which definitions and populations differed considerably. Our purpose was to evaluate the incidence and importance of recurrent ischemia after administration of thrombolytic therapy. We studied patients enrolled in the Thrombolysis and Angioplasty in Myocardial Infarction studies. Patients were enrolled into 5 studies with similar entry criteria; 552 patients were treated with tissue plasminogen activator (t-PA), 293 were treated with urokinase, and 385 received both thrombolytic agents. Recurrent ischemia was defined as symptoms in association with electrocardiographic changes; reinfarction was defined as a reelevation of creatine kinase myocardial band isoenzyme in an appropriate clinical setting. Both recurrent ischemia and reinfarction occurred in 42 patients (3.4%), recurrent ischemia alone occurred in 226 (18%), whereas neither occurred in 964 (78%). Although baseline characteristics were similar among the 3 groups, in-hospital cardiac events (total 73 deaths, 253 heart failure episodes) were not: in-hospital mortality in patients with reinfarction was 21%; with recurrent ischemia, 11%; and with neither event, 4% (p < 0.0001). The in-hospital heart failure rate of patients with reinfarction was 50%; with recurrent ischemia alone, 31%; and with neither event, 17% (p < 0.0001). As expected, median in hospital costs were highest in patients with reinfarction ($26,802), intermediate for those with recurrent ischemia alone ($18,422), and lowest in patients with neither event ($15,623). Recurrent myocardial ischemia after thrombolytic therapy is a frequent, important, and expensive adverse clinical outcome, making it a critical target for therapeutic intervention. PMID- 7484853 TI - Prognostic value of training-induced change in peak exercise capacity in patients with myocardial infarcts and patients with coronary bypass surgery. AB - An inverse association between mortality and exercise capacity has been demonstrated previously in patients with coronary artery disease. Physical training generally increases exercise capacity. Only 1 study investigated the prognostic value of exercise capacity after training, but only in a limited number of patients. No data are available on the relation between mortality and the change in exercise performance with training. Peak oxygen uptake (VO2) was measured before and after a 3-month, predominantly dynamic training period in 417 patients with coronary artery disease. Apart from peak VO2, several patient characteristics, risk factors for cardiovascular disease, and exercise data were considered in a Cox proportional-hazards model. Peak VO2 had increased by 33% after the training period. During the total follow-up of 2,583 patient-years, 37 patients died. The cause of death was cardiovascular in 21. The prognostic value of peak VO2 was higher after training than before training, even after adjustment for age and other significant covariates. Cardiovascular mortality decreased more with greater increases in peak VO2 after training. The relative hazard rate of 0.98 indicates that a 1% greater increase in peak VO2 after training would be associated with a decrease in cardiovascular mortality of 2%. No differences in prognostic value and in training effects were observed between patients with myocardial infarcts and patients after coronary bypass grafting. Peak VO2, evaluated after a physical training program, and its change in response to training are independent predictors for cardiovascular mortality in patients with coronary artery disease. PMID- 7484854 TI - Enhanced hepatic uptake of thallium-201 in patients with severe narrowing of the right coronary artery. AB - To assess the hepatic uptake of thallium-201 after exercise treadmill testing and to investigate whether hepatic uptake of thallium-201 may be a useful marker of right coronary artery (RCA) disease, 43 patients were studied: 17 with RCA disease (9 with 1-vessel disease, 8 with multivessel disease including the RCA), 8 with left coronary system disease alone, and 18 with a low probability (< 5%) of coronary disease. All subjects were studied with symptom-limited exercise and redistribution thallium-201 single-photon emission computed tomographic (SPECT) scintigraphy. Two indexes of hepatic uptake were derived: a liver-to-heart ratio after stress, and a stress-to-rest hepatic ratio. The low-probability group had a liver/heart ratio of 0.48 +/- 0.02. In the group with RCA disease alone, liver/heart ratio was 1.29 +/- 0.20 (p < 0.005 vs low-probability group). Patients with multivessel coronary artery disease involving the RCA had a ratio of 1.19 +/- 0.16 (p < 0.005 vs low-probability group), and patients with only left coronary system disease had a liver/heart ratio of 0.87 +/- 0.15 (p < 0.05 vs low-probability group). The stress/rest ratio of the low-probability group was 0.83 +/- 0.04. Patients with RCA disease alone had a stress/rest ratio of 1.49 +/ 0.25 (p < 0.05 vs low-probability group), and patients with multivessel disease involving the RCA had a stress/rest ratio of 1.16 +/- 0.08 (p < 0.005 vs low probability group).(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7484855 TI - A meta-analysis of randomized trials comparing coronary artery bypass grafting with percutaneous transluminal coronary angioplasty in multivessel coronary artery disease. AB - We performed a meta-analysis of randomized trials that compared percutaneous transluminal coronary angioplasty (PTCA) with coronary artery bypass graft (CABG) surgery in patients with multivessel coronary artery disease. The outcomes of death, combined death, and nonfatal myocardial infarction (MI), repeat revascularization, and freedom from angina were analyzed. The overall risk of death and nonfatal MI was not different over a follow-up of 1 to 3 years (CABG:PTCA odds ratio [OR] 1.03, 95% confidence interval 0.81 to 1.32, p = 0.81). Patients randomized to CABG tended to have a higher risk of death or MI in the early, periprocedural period (OR 1.33, p = 0.091), but a lower risk in subsequent follow-up (OR 0.74, p = 0.093). CABG patients were much less likely to undergo another revascularization procedure (p < 0.00001), and were more likely to be angina free (OR 1.57, p < 0.00001). Thus, CABG and PTCA patients have similar overall risks of death and nonfatal MI at 1 to 3 years of follow-up, but relative risk differences in mortality of up to 25% cannot be excluded. CABG patients have significantly less angina and less repeat revascularization than PTCA patients. PMID- 7484856 TI - The limited efficacy of exercise radionuclide ventriculography in assessing prognosis of women with coronary artery disease. AB - Increasing numbers of women are undergoing stress testing for coronary artery disease evaluation. Limited study is available as to its efficacy in women. Four hundred nineteen patients with coronary artery disease (74 women and 345 men) referred for exercise radionuclide ventriculography between 1979 and 1986 were evaluated in a prospective cohort evaluation with 5-year follow-up. Exercise radionuclide ventriculographic variables were analyzed and compared between women and men. The prognostic efficacy of exercise radionuclide ventriculography was assessed separately for women and men among patients with coronary artery disease by Kaplan-Meier cumulative survival curves, univariate Cox regression analyses, and hierarchical stepwise Cox regression analyses. Overall, women demonstrated higher resting and peak left ventricular ejection fraction response to exercise than men. Ninety-six of 419 patients (23%) had cardiac events at 5-year follow up. Although left ventricular response to exercise conveyed prognostic information in the combined and male populations (multivariate hierarchical analyses chi-square 11, p = 0.001 for delta left ventricular ejection fraction and chi-square 10, p = 0.002 for worsening exercise wall motion score), these variables were not found to be prognostically useful in women. Women with coronary artery disease demonstrated a worsened functional status, evidenced by greater compromise of exercise capacity, despite having less extensive anatomic disease than their male counterparts. We conclude that sex-related differences in left ventricular response to exercise limit the prognostic utility of exercise ventriculography in women with coronary artery disease. PMID- 7484857 TI - American College of Emergency Physicians information paper: chest pain units in emergency departments--a report from the Short-Term Observation Services Section. PMID- 7484858 TI - Electrophysiologic profile and efficacy of intravenous dofetilide (UK-68,798), a new class III antiarrhythmic drug, in patients with sustained monomorphic ventricular tachycardia. Dofetilide Arrhythmia Study Group. AB - There is increasing evidence that class III antiarrhythmic agents may be superior to class I agents for the long-term treatment of life-threatening ventricular tachyarrhythmias. This open study evaluated the acute electrophysiologic effects, antiarrhythmic efficacy, and safety of different doses of intravenous dofetilide, a new class III drug, in 50 patients with sustained monomorphic ventricular tachycardia inducible by programmed electrical stimulation who had previously been unsuccessfully treated with 0 to 7 (median 3) other drugs. Intravenous dofetilide was administered over 60 minutes at the following dose levels: 1.5, 3.0, 6.0, 9.0, and 15.0 micrograms/kg. Significant class III activity was apparent at doses of 3.0 to 15.0 micrograms/kg, as evidenced by dose-related prolongation of the QTc interval by 13.4% to 14.2%, ventricular effective refractory period by 7.9% to 20.6%, and ventricular functional refractory period by 7.3% to 25.0%. The corresponding mean +/- SD plasma dofetilide concentrations ranged from 1.45 +/- 0.52 to 6.48 +/- 1.31 ng/ml. There was no evidence of reverse use-dependence. At these electrophysiologically active dose levels, intravenous dofetilide suppressed (complete response) or slowed (partial response) inducible ventricular tachycardia in 17 of 41 patients (41%) compared with 0 of 9 patients receiving only 1.5 micrograms/kg. The response rate was fairly uniform among the groups receiving 3.0, 6.0, 9.0, and 15.0 micrograms/kg. Intravenous dofetilide was hemodynamically well tolerated. Torsades de pointes (which was self-limiting) developed in only 1 patient, who was allocated to receive 15.0 micrograms/kg. There were no other proarrhythmic episodes or serious adverse effects. Further evaluation of the therapeutic potential of dofetilide in the management of life-threatening ventricular arrhythmias is justified. PMID- 7484859 TI - Effects of enalapril on heart rate variability in patients with congestive heart failure. AB - Congestive heart failure (CHF) is characterized by sympathetic activation and parasympathetic withdrawal, and the magnitude of sympathoneural activation is associated with adverse outcome. Angiotensin-converting enzyme inhibitor therapy has been shown to reduce mortality and improve prognosis in patients with CHF, but whether this therapy improves cardiac autonomic control is not well known. This double-blind, placebo-controlled, crossover study examines the effects of enalapril on autonomic control in 12 patients with mild to moderate CHF by heart rate variability analysis. Compared with placebo, enalapril increased the SD of all normal RR intervals (SDNN) from 39 +/- 13 to 48 +/- 15 ms (p < 0.01), the SD of the average RR intervals for all 5-minute segments from 33 +/- 12 to 42 +/- 15 ms (p < 0.01), and the mean of the SDs of all RR intervals for all 5-minute segments (SDNN index) from 19 +/- 5 to 23 +/- 6 ms (p < 0.01). The root-mean square successive differences and the percent differences between adjacent RR intervals > 50 ms were also increased from 17 +/- 8 to 21 +/- 8 ms (p < 0.01) and from 1.1 +/- 2.1 to 2.8 +/- 2.9% (p < 0.05). In addition, total, low-frequency, and high-frequency power were increased from 560 +/- 349 to 786 +/- 504 ms2/Hz (p < 0.01), from 125 +/- 107 to 179 +/- 135 ms2/Hz (p < 0.01), and from 46 +/- 32 to 94 +/- 78 ms2/Hz (p < 0.01), respectively.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7484860 TI - Right subscapular artery catheterization for balloon valvuloplasty of critical aortic stenosis in infants. AB - This study was performed to evaluate the utility and safety of catheterizing the right subscapular artery for balloon valvuloplasty of critical aortic stenosis in infants. Twenty-one patients, age 20 days to 17 months, underwent attempted valvuloplasty through the surgically exposed right subscapular artery. Five or 7Fr catheters with balloon diameters of 7 to 10 mm were used. Valvuloplasty was successfully performed using this approach in 11 patients. In 2 other patients, the subscapular artery would not accommodate the balloon angioplasty catheter (7Fr), and the arteriotomy was extended into the axillary artery. In these 13 patients, the peak systolic pressure gradient across the aortic valve was decreased from 85 +/- 23 to 33 +/- 7 mm Hg. Moderate aortic regurgitation developed in 3 patients. In the remaining 8 patients, valvuloplasty could not be performed through the right subscapular artery. In 2 patients, the right subclavian artery was anomalous and led to the descending aorta. In 6 small patients, no catheter could be advanced across the aortic valve. In 1 of these patients, a guidewire perforated a coronary sinus of Valsalva causing death. Overall, valvuloplasty using the right subscapular arterial approach was successful in 13 of 19 infants (68%) with normal right subclavian arteries, including all 10 such patients weighing > or = 5.5 kg. No clinically significant peripheral vascular complications or brachial plexus injuries occurred. Thus, the right subscapular arterial approach is an alternative route to be considered when planning balloon aortic valvuloplasty in infants. PMID- 7484862 TI - Effects of ionic and nonionic contrast media on in vitro and in vivo platelet activation. AB - An increased incidence of thromboembolic events has been associated with nonionic contrast media. Although it has been demonstrated that nonionic contrast media cause significant platelet activation in vitro, there is no reported evidence of contrast-induced platelet activation in vivo. This study investigated the effect of a nonionic contrast agent, iohexol (Omnipaque), versus an ionic contrast agent, diatrizoate meglumine+diatrizoate sodium (MD-76), on both in vitro and in vivo platelet activity. Blood from normal subjects (n = 12) was incubated in vitro for 15 to 60 seconds with varying proportions of contrast media or saline solution, and analyzed by flow cytometry for markers of platelet degranulation (P selectin) and platelet GPIIb-IIIa receptor activation (PAC-1). In vivo activation after Omnipaque was measured in patients undergoing cardiac catheterization in peripheral blood before and after contrast (n = 5), and in coronary sinus blood before and after left coronary injections (n = 8). In vitro data showed that Omnipaque incubated for 60 seconds with whole blood at ratios of 1:1 resulted in a greater percentage of platelets expressing P-selectin (median/range) than did MD-76 (68%/38% to 90% vs 17%/1% to 53%, respectively, p < 0.01). Similarly at a contrast: blood ratio of 1:3, Omnipaque caused greater P-selectin expression than did MD-76 (49%/19% to 83% vs 3%/0% to 5%, respectively, p < 0.05). There was no significant increase in platelet degranulation observed with concentrations of contrast more dilute than 1:3. In addition, there was significantly less activation seen even at contrast:blood ratios of 1:1 when exposure time was < 60 seconds (p < 0.01).(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7484861 TI - Dynamic three-dimensional echocardiographic assessment of intracardiac blood flow jets. AB - Transthoracic dynamic 3-dimensional reconstruction of the heart with tissue depiction has been proved to be feasible when using various methods of data acquisition. The same method can theoretically be applied to color Doppler flows to generate dynamic 3-dimensional images of intracardiac blood flow jets. To explore the feasibility of this approach, we studied 41 patients with various valvular disorders or intracardiac shunts. We acquired sequential 2-dimensional images along with color Doppler information using rotational scanning from a transthoracic or a subcostal window. Images were digitized and processed for 3 dimensional reconstruction using dedicated software. After adequate segmentation, the flow jets were displayed in 3 dimensions in a gray scale format. With use of this approach, 3-dimensional reconstruction of color Doppler flows was possible in all but 1 patient. Still frames allowed immediate appreciation of the shape of the jets, their location in the cardiac chambers, and their size related to that cavity. Dynamic display was even more striking by showing the flow in real time. Dynamic 3-dimensional images enabled visualization of flow jets in projections not available in conventional color flow Doppler, looking directly at the views of shunt and regurgitant flows, and also permitted 3-dimensional delineation of flow convergence zones. We conclude that dynamic visualization of various intracardiac flows in 3 dimensions using transthoracic echocardiography is possible. It provides a better understanding of the shape and size of the jets, and can potentially aid in flow quantification by displaying the actual shape of flow convergence regions. PMID- 7484863 TI - Unexpected death in coronary artery disease. PMID- 7484864 TI - In vivo assessment by intravascular ultrasound of enlargement in saphenous vein bypass grafts. PMID- 7484865 TI - Cardiac rehabilitation using simultaneous voice and electrocardiographic transtelephonic monitoring. PMID- 7484866 TI - Effects of the antiestrogen tamoxifen on low-density lipoprotein concentrations and oxidation in postmenopausal women. AB - Our study demonstrates that tamoxifen, when administered to postmenopausal women at a conventional dosage, reduces LDL levels and protects LDL from oxidation. The protective effect of tamoxifen against the development of breast cancer in women considered at risk is being investigated in a placebo-controlled trial sponsored by the National Institutes of Health. Whether tamoxifen also protects against the development of cardiovascular disease in this trial is also of considerable interest. PMID- 7484868 TI - Breath acetone in congestive heart failure. AB - To summarize, CHF predisposes to postabsorptive ketosis in relation to the severity of venous congestion. A simple and fully noninvasive measurement of breath acetone may add to the diagnostic assessment of patients with CHF. PMID- 7484867 TI - Age- and anesthesia-related changes in accessory pathway conduction in children with Wolff-Parkinson-White syndrome. AB - The anterograde effective refractory period of the accessory pathway is age dependent in pediatric patients with the WPW syndrome. Thus, age should be considered when developing electrophysiologic criteria for the risk of hypotensive arrhythmias in these patients. In addition, general anesthesia must also be considered in interpreting age-related changes in the anterograde APERP, especially in children. PMID- 7484870 TI - Use of the Terumo SP catheter system for crossing the pulmonary valve in infants with critical pulmonary valve stenosis. AB - In short, this method (the Terumo SP hydrophilic-polymer coated microcatheter system) has proved to be safe, effective, and flexible; it has allowed us to cross critically stenotic valves when standard methods have failed, enabling us to accomplish balloon valvuloplasty with good results in patients who might have otherwise required surgical intervention because of inability to cross the pulmonary valve. PMID- 7484869 TI - Effect of long-term angiotensin-converting enzyme inhibition on vascular function in patients with chronic congestive heart failure. AB - The present study demonstrates that peripheral vasodilatory capacity is impaired in patients with chronic congestive heart failure not treated with aspirin, but preserved in patients taking aspirin. This decreased peripheral vasodilatory capacity can be restored by chronic angiotensin-converting enzyme inhibition, indicating that locally acting cyclooxygenase-dependent factors contribute to peripheral vasoconstriction in chronic congestive heart failure. PMID- 7484871 TI - Influence of bidirectional superior cavopulmonary anastomosis on pulmonary arterial growth. AB - Right-sided BSCA provides for satisfactory pulmonary arterial growth in infants and children with complex congenital heart defects, and it could enhance the growth of a small right pulmonary artery. The growth of the left pulmonary artery, particularly in younger patients, needs close attention to confirm the safe role of BSCA in long-term palliation. PMID- 7484872 TI - Gender-related differences on signal-averaged electrocardiograms in healthy subjects. PMID- 7484873 TI - The effects of emotions on short-term power spectrum analysis of heart rate variability . AB - In summary, this work extends previous findings by demonstrating that anger produces a sympathetically dominated power spectrum, whereas appreciation produces a power spectral shift toward MF and HF activity. Results suggest that positive emotions lead to alterations in HRV, which may be beneficial in the treatment of hypertension and in reducing the likelihood of sudden death in patients with congestive heart failure and coronary artery disease. PMID- 7484874 TI - Effect of menopause on left ventricular filling in 50-year-old women. PMID- 7484875 TI - Comparison of ST-segment/heart rate slope analysis with standard ST-segment measurement criteria to outcome of exercise thallium-201 imaging. AB - We compared standard ST-segment analysis and ST/HR slope analysis of exercise ECG studies with reference to outcome of exercise thallium-201 studies in 341 patients. Sensitivity was significantly better using ST/HR slope compared with standard ST analysis. Specificity was not significantly different. PMID- 7484877 TI - Role of transesophageal echocardiography in the management of patients with bronchogenic carcinoma invading the left atrium. AB - TEE played a significant role in the management of 2 patients by defining the extent of cardiac invasion. We believe TEE would be helpful in patients presenting with thoracic tumors believed to be encroaching on cardiac structures, where consideration is being given to surgical resection of the tumor. PMID- 7484876 TI - Identical twins with arrhythmogenic right ventricular dysplasia. PMID- 7484878 TI - Prevention of venous thrombosis after coronary artery bypass surgery (a randomized trial comparing two mechanical prophylaxis strategies). AB - Although venous thrombosis may occur often after coronary artery bypass grafting, prophylaxis with low-dose heparin is rarely used due to the risk of bleeding. Therefore, we compared the efficacy of 2 mechanical regimens of prophylaxis against deep vein thrombosis (DVT). Consecutive patients undergoing coronary artery bypass without concomitant valve surgery or coronary endarterectomy were randomized to either a more intensive regimen of intermittent pneumatic compression (IPC) plus graduated compression stockings (GCS) versus standard compression stockings alone. Of 611 patients screened, 184 were excluded due to peripheral vascular disease, postoperative intraaortic balloon support, or immediate postoperative anticoagulation. An additional 83 patients refused consent, leaving 172 in each prophylaxis group. The primary study end point was DVT diagnosed by a predischarge leg ultrasound examination performed on postoperative days 4 to 6. Of 344 patients enrolled, 330 (96%) underwent predischarge ultrasonography. DVT was detected in 19% of patients assigned to IPC plus stockings versus 22% assigned to GCS alone (95% confidence interval for the difference, -11% to 6%, p = 0.62). The addition of IPC did not add significant incremental benefit to GCS alone for DVT prophylaxis among patients undergoing coronary artery bypass surgery. PMID- 7484879 TI - Frequency of "optimal anticoagulation" for acute myocardial infarction after thrombolysis with front-loaded recombinant tissue-type plasminogen activator and conjunctive therapy with recombinant hirudin (HBW 023). ALKK Study Group. AB - This retrospective analysis reviewed 183 patients with acute myocardial infarction who were given front-loaded recombinant tissue-type plasminogen activator (rt-PA) and r-hirudin (HBW 023) in 1 of 4 dose groups (bolus dose of 0.07, 0.1, 0.2, or 0.4 mg/kg, followed by an infusion of 0.05, 0.06, 0.1, or 0.15 mg/kg/hour over 48 hours). Activated partial thromboplastin time (aPTT) levels were determined at baseline and at 4, 8, 12, 16, 20, 24, 32, 40, and 48 hours. Of the 178 patients with r-hirudin treatment for > or = 12 hours, anticoagulation was optimal in 55.1% (all aPTTs > 2 x baseline), suboptimal in 33.7% (lowest aPTT > 1.5 but < 2 x baseline), and inadequate in 11.2% (> or = 1 aPTT but < 1.5 x baseline). Optimal anticoagulation was observed more frequently in the higher dose groups (dose 1, 15%; dose 2, 44.4%; dose 3, 63.4%; dose 4, 73.4%; p for trend < 0.0001). Patency (according to Thrombolysis in Myocardial Infarction trial grade 2 or 3) of the infarct artery after 36 to 48 hours was higher in the group with optimal anticoagulation compared with those with suboptimal or inadequate anticoagulation: 97.9%, 88.4%, and 85%, respectively (p = 0.03 optimal vs suboptimal or inadequate anticoagulation). In conclusion, r-hirudin in a dose of 0.1 or 0.15 mg/kg/hour achieves an optimal anticoagulation in about 63% or 74% of patients, which is associated with an enhanced patency 24 to 48 hours after rt PA. A subsequent study revealed that this effective anticoagulation may be accompanied by an increased risk of severe bleeding complications after thrombolysis. PMID- 7484880 TI - A symposium: from hypertension to heart failure: endothelial dysfunction and structural changes in cardiovascular disease. Introduction. PMID- 7484881 TI - Effect of chronic angiotensin-converting enzyme inhibition on endothelial function in patients with chronic heart failure. AB - Chronic heart failure is associated with neurohumoral activation and alterations of the peripheral circulation and skeletal muscle. Several mechanisms are involved in the impaired peripheral perfusion, including increased sympathetic tone and increased vascular stiffness. Recently, data suggest an important role of the endothelium for perfusion of skeletal muscle in heart failure. Endothelium dependent dilation of resistance vessels is blunted in patients with severe chronic heart failure and may be involved in the impaired reactive hyperemia in these patients. In conductance vessels, flow-dependent dilation and the nitroglycerin-induced dilator response is attenuated in congestive heart failure as compared to normal subjects, indicating both endothelial dysfunction and a defect of smooth muscle relaxation. Recent data suggest that angiotensin converting enzyme (ACE) inhibitors can improve endothelial function of resistance vessels, reduce serum level of the soluble endothelial (vascular cell) adhesion molecule (VCAM-1) and, in addition, improve peripheral vascular function by reducing or limiting the influence of cyclo-oxygenase-dependent vasoconstricting factor(s). It is conceivable that these beneficial effects of chronic ACE inhibition are due, in part, to blockade of bradykinin degradation by the ACE and the increased endothelial synthesis of prostaglandins and/or the release of nitric oxide by enhanced tissue levels of bradykinin. PMID- 7484882 TI - Effect of aging and chronic angiotensin I converting enzyme inhibition on the endothelial function of the mesenteric arterial bed of the rat. AB - Endothelial function was studied in perfused mesenteric arterial bed preparations removed from 6, 12, 24, or 30 month-old, male, normotensive rats. Half of the animals were treated with a hypotensive dose of an angiotensin I converting enzyme (ACE) inhibitor (perindopril, 1 mg/kg/day, p.o.) from 6 months onwards. Disruption of endothelium produced a fall in baseline perfusion pressure in all animals except 30-month old controls, suggesting that the maintenance of baseline tone by endothelial vasoconstrictor(s) is impaired with age. Physiologic antagonism of norepinephrine-induced vasoconstriction by the muscarinic agonist carbachol was impaired in 30-month old controls. Norepinephrine-induced vasoconstriction increased following disruption of endothelium, suggesting that norepinephrine-induced release of endothelium-derived relaxing factor attenuates vasoconstriction. This mechanism was impaired in 30-month old controls. Chronic ACE inhibition postponed the age-related decrease in vasodilator and vasoconstrictor endothelial functions, either directly or indirectly via the hypotensive action of such treatment. PMID- 7484883 TI - Angiotensin-converting enzyme inhibitors reduce neointimal thickening and maintain endothelial nitric oxide function in rabbit carotid arteries. AB - Application of periarterial collars induced atheroma-like lesions in the carotid arteries of normocholesterolemic rabbits. Vessel segments taken from the mid region of the collar (cuffed region) and control regions of the same artery were studied at 7 days after surgery. A group of placebo rabbits was provided ad libitum with regular tap water, and treated animals were supplied for 14 days (beginning 7 days before collar application) with water containing perindopril (daily intake of approximately 0.3 mg/kg). Perindopril treatment reduced plasma angiotensin-converting enzyme (ACE) activity by 88% but did not significantly alter arterial blood pressure or heart rate. The sensitivity of arterial rings to angiotensins I and II did not differ between control and cuffed arteries in either placebo or perindopril-treated rabbits, but in rings from both groups of rabbits the sensitivity to the vasoconstrictor action of serotonin was higher in the cuffed segments, as in previous studies. In addition, in placebo rabbits the endothelium-dependent vasorelaxant response to acetylcholine (which results from the release of nitric oxide) was weaker in cuffed arteries than in controls, whereas in the perindopril-treated animals, this impairment of relaxation was restored to the extent that, in cuffed vessels, it was no longer significantly different from the controls. Similar results were obtained in rabbits treated with another ACE inhibitor (ramipril). In contrast, acute treatment with the metabolite, perindoprilat, in vitro (1.0 microM) did not alter the response to acetylcholine in control or cuffed rings from placebo rabbits. Morphologically, vessel segments taken from the center of the cuffed artery of placebo rabbits showed a thickened intima with marked smooth muscle cell proliferation.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7484884 TI - Vascular and myocardial protective effects of converting enzyme inhibition in experimental heart failure. AB - Systemic vasoconstriction due to stimulation of the sympathetic and renin angiotensin-aldosterone systems is a hallmark of heart failure and this is accompanied by impaired endothelium-dependent relaxations at the level of large arteries. This study investigated, in a rat model of heart failure, whether such an endothelial dysfunction also exists at the level of the resistance artery, and whether this is associated with morphologic changes, as well as the effects of chronic treatment with the angiotensin-converting enzyme inhibitor perindopril (2 mg/kg/day). After 12 months, arterial pressure, left ventricular (LV) end diastolic pressure (LVEDP), and LV dP/dt were measured in anesthetized rats. Responses to acetylcholine and nitroprusside were determined in isolated and perfused mesenteric artery segments (diameter: 280 +/- 15 microns). After fixation, vessel diameter, media cross-sectional area, and media collagen and elastin densities were measured by image analysis. After 12 months, untreated rats showed signs of heart failure, i.e., reduced LV dP/dt, and increased LVEDP, heart weight/body weight, LV cavity circumference, and myocardial collagen density. In mesenteric vessels the endothelium-dependent vasodilator response to acetylcholine was impaired, whereas the response to the nitric oxide donor nitroprusside was unaffected. Heart failure did not affect vascular morphological parameters. Perindopril decreased blood pressure and LVEDP without any modification of LV dP/dt, and prevented cardiac remodeling. At the vascular level, perindopril improved the response to acetylcholine and reduced media cross sectional area and collagen density without affecting internal vessel diameter or elastin density. Thus, heart failure decreases endothelium-dependent vasodilator response to acetylcholine without modification of vessel structure.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7484885 TI - Endothelium-derived relaxing factors and converting enzyme inhibition. AB - Endothelial cells can produce at least 3 substances which cause relaxation of vascular smooth muscle: (1) endothelium-derived nitric oxide (NO, which is secreted not only toward the underlying vascular smooth muscle but also into the blood vessel lumen). NO also has a physiological role at the interface between the endothelial cells and the blood content; in particular, NO inhibits the adhesion of platelets and leukocytes to the endothelium. (2) Endothelium-derived hyperpolarizing factor, presumably a labile metabolite of arachidonic acid formed through the P-450 pathway, which appears to act on smooth muscle by being one of the few physiologic openers of the potassium channels. (3) Prostacyclin, which can be considered as an endothelium-derived relaxing substance, given its vasodilator activity and its primarily endothelial origin. One of the main factors modulating the release of these EDRFs is the shear stress of blood on the arterial wall, which explains why flow-induced vasodilation is endothelium dependent in the intact organism. The peptide bradykinin is a potent stimulus for EDRF release. The normal lifespan of an adult human endothelial cell is some 30 years, after which aging takes its toll and the cells must be replaced. The regenerated cells lose some of their ability to release EDRF, in particular in response to platelet aggregation and thrombin. Finally, in hypertension and atherosclerosis, a decrease in endothelium-dependent relaxation is obvious in response to a variety of stimuli. All converting enzyme inhibitors tested so far share a potentiating effect on endothelium-dependent relaxation to bradykinin, and augmented local production of bradykinin may help to explain the acute vasodilator properties of these compounds. PMID- 7484886 TI - Structural changes in cardiovascular disease. AB - The growing emphasis on structural alterations in the heart and vasculature as a critical common pathway toward morbidity and mortality in a wide variety of cardiovascular diseases has opened the door to new interventions that might fundamentally affect these processes. Converting enzyme inhibitors, because of their demonstrable effect on both left ventricular and vascular remodeling, are attractive agents to alter the natural history of these diseases. Whether the effect of converting enzyme inhibitors is mediated by a reduction of angiotensin II, an increase in bradykinin, or other local growth factors remains to be established. Other agents may share or complement the actions of angiotensin converting enzyme (ACE) inhibitors in inhibiting these unwanted remodeling processes. It is clear that the future of therapy for chronic cardiovascular diseases must take into consideration the structural abnormalities that in the past have been given inadequate attention. New and more targeted therapy to alter specific structural changes may profoundly affect the natural history of these diseases. PMID- 7484887 TI - Regression of media-to-lumen ratio of human subcutaneous arteries and left ventricular hypertrophy during treatment with an angiotensin-converting enzyme inhibitor-based regimen in hypertensive patients. AB - A total of 25 patients with newly diagnosed or poorly controlled essential hypertension were randomly selected from a larger group referred to hospital. Treatment was initiated with perindopril (4-8 mg orally). If normotension was not achieved, isradipine (5-10 mg orally) was added and, if necessary, hydralazine was added. Before treatment and at the end of a 9-month period of normotension (diastolic blood pressure < or = 90 mm Hg), 24-hour blood pressure and echocardiographic measurements were performed and resistance artery structure was determined. A total of 20 age- and sex-matched normotensives were used as controls. During antihypertensive treatment, mean blood pressure was reduced from 128 +/- 11 to 103 +/- 6 mm Hg. Left ventricular mass was reduced from 300 +/- 76 to 198 +/- 54 g. The media:lumen ratio of the resistance arteries decreased from 9.8 +/- 2.6% to 7.8 +/- 1.9%; control subjects exhibited a media:lumen ratio of the same magnitude (7.9 +/- 2.0%). Results indicate that a perindopril-based regimen is extremely efficient in normalizing resistance artery and cardiac ventricular structures within one year of treatment. The impact of these findings on the excess cardiovascular morbidity and mortality in arterial hypertension still remains to be demonstrated. PMID- 7484888 TI - Chronic low-dose treatment with perindopril improves cardiac function in stroke prone spontaneously hypertensive rats by potentiation of endogenous bradykinin. AB - We investigated the effect of chronic angiotensin-covering enzyme (ACE) inhibitor treatment on functional and biochemical cardiac parameters in stroke-prone spontaneously hypertensive rats (SHRsp). Animals were treated prenatally and, subsequently, up to the age of 20 weeks with the ACE inhibitor perindopril (0.01 and 1 mg/kg per day). The contribution of endogenous bradykinin potentiation to the actions of the ACE inhibitor was assessed by co-treatment with the bradykinin B2-receptor antagonist, icatibant (500 micrograms/kg/day s.c.), from 6 to 20 weeks of age and by measurement of myocardial prostacyclin and cyclic guanosine monophosphate (GMP) concentrations. Chronic high-dose treatment with perindopril attenuated the development of hypertension and left ventricular hypertrophy while low-dose perindopril treatment had no effect on these parameters. However, low dose perindopril improved cardiac function of isolated perfused hearts as demonstrated by an increasing left ventricular pressure and dp/dtmax without change in heart rate. Low-dose perindopril further reduced lactate concentrations and the enzymatic activities of lactate dehydrogenase and creatine kinase in the coronary venous effluent and increased tissue concentrations of glycogen, adenosine triphosphate, and creatine kinase in the myocardium. Concomitant chronic bradykinin receptor blockade abolished all ACE inhibitor-induced effects on cardiac function and metabolism. Cardiac prostacylin concentrations were 3 fold elevated in perindopril-treated animals when compared to vehicle-treated controls, while cardiac cyclic GMP concentrations remained unchanged. Our data demonstrate that chronic ACE inhibitor treatment can improve cardiac function and metabolism independently of the antihypertensive and antihypertrophic drug actions by potentiation of endogenous bradykinin. PMID- 7484889 TI - Disparate effects of antihypertensive drugs on large artery distensibility and compliance in hypertension. AB - Distensibility and compliance are large artery properties, that may be important in cardiovascular disease. Distensibility is a determinant of the pulsatile stress on the vessel wall and is thought important in ageing and atherosclerotic disease. Compliance reflects the buffering capacity of the arteries and is a major determinant of the afterload on the heart. In hypertension large arteries are getting stiffer, resulting in a decreased distensibility and compliance. Decrease in blood pressure by itself can improve large artery properties. Despite a decrease in blood pressure, not all antihypertensive drugs improve large artery properties. Compliance is improved by firstline antihypertensive drugs such as angiotensin-converting enzyme (ACE) inhibitors, calcium antagonists, beta blockers with vasodilating properties, selective beta 1-blockers and some diuretics. Recent data suggest that ACE inhibitors such as perindopril improve distensibility and compliance of large arteries more than diuretics such as amiloride/hydrochlorothiazide. Apart from the indirect effect (decrease in blood pressure), this makes a direct effect of perindopril on large artery properties very likely. ACE inhibitors such as perindopril decrease the afterload on the heart more than diuretics such as amiloride/hydrochlorothiazide by both a more pronounced decrease in systemic vascular resistance and an increase in large artery compliance. PMID- 7484890 TI - Vascular wall thickness in hypertension: the Perindopril Regression of Vascular Thickening European Community Trial: PROTECT. AB - A high prevalence of increased intima/media thickness of the arterial wall has been documented in hypertension. These alterations in vascular wall structure may be potent determinants for the promotion of the development of atherosclerosis. Direct histologic data from animal models of hypertension, and indirect data from hypertensive patients, have demonstrated a marked regression of increased intima/media thickness by angiotensin-converting enzyme (ACE) inhibition. Long term effects of ACE inhibition on structural wall changes in humans have not been examined. Therefore, a multicenter, randomized, double-blind European trial was designed to compare the effects of the ACE inhibitor perindopril and the diuretic hydrochlorothiazide in slowing or reversing progression of increased intima/media thickness of carotid and femoral arteries in hypertensive patients. A total of 800 patients at 17 clinical centers in 7 European countries, aged 35-65 years, with hypertension and ultrasonographically proven intima/media thickness > or = 0.8 mm of the common carotid artery will be randomly assigned to receive in a double-blind fashion either perindopril or hydrochlorothiazide and will be followed for 24 months. High resolution duplex sonography will be used to quantify intima/media thickness at baseline and twice a year during follow-up. A change of 0.1 mm of intima/media thickness from baseline is considered to be detectable, and the standard deviations of the changes from baseline are expected not to be higher than 0.2 mm. The primary endpoint of the study is the comparison of changes in intima/media thickness of the common carotid artery.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7484891 TI - Comparison of mortality from acute myocardial infarction in patients receiving anistreplase with those not receiving thrombolysis. AB - Within 1 year, 434 patients were admitted to 14 hospitals with suspected acute myocardial infarction (AMI) < or = 4 hours after the onset of symptoms. Group A consisted of 171 patients (39%) treated with thrombolysis, and group B consisted of 263 patients (61%) with contraindications. Patients in group A more likely had a "definite AMI" (92%; group A1) than patients in group B (67%; group B1). Group B1 had 277 contraindications (1.6/per patient) with increased risk for life threatening bleeding being the most frequently recorded at admission. The in hospital mortality in group A1 was 7% (11 of 158) and in group B1, 27% (47 of 177) (p < 0.0001). Age and type of therapy (thrombolysis or no thrombolysis) were identified as independent predictors of increased mortality (p < 0.0001 and < 0.05, respectively). Thus, although most patients with an AMI are excluded from thrombolytic therapy because of contraindications, our data suggest that their in hospital mortality is unexpectedly high. Further evaluation of this group of patients is warranted to define the impact of contraindications as an independent factor of mortality. PMID- 7484892 TI - Value of rescue percutaneous transluminal coronary angioplasty following unsuccessful thrombolytic therapy in patients with acute myocardial infarction. Thrombolysis in Myocardial Infarction Investigators. AB - Rescue percutaneous transluminal coronary angioplasty (PTCA) has been advocated as a mechanical method to achieve reperfusion in instances where the myocardial infarct artery remains occluded after thrombolytic therapy. Most prior reports of rescue PTCA have been observational and analyses of value have been inconclusive. To evaluate the benefit of rescue PTCA, we studied 133 patients enrolled in the Thrombolysis in Myocardial Infarction Phase I Open Label and Phase II trials who had an occluded infarct-related artery after thrombolytic therapy. According to protocol, 100 consecutive patients had no rescue PTCA performed (no rescue group), and 33 consecutive patients underwent protocol-directed rescue PTCA (rescue group). The 2 cohorts were compared for clinical and angiographic outcome. Baseline characteristics of the 2 groups were similar. Rescue PTCA was attempted in each rescue group patient and was successful in 82%. At 21 days the mortality rate was 12% in the rescue group and 7% in the no rescue group (p = NS). Failed rescue PTCA was associated with a mortality of 33%. Reinfarction occurred in 6% of patients in the rescue group, and in 5% of those in the no rescue group (p = NS). At 21 days, mean left ventricular ejection fraction was 51 +/- 13% in the rescue group and 48 +/- 12% in the no rescue group (p = NS). We conclude that the routine use of rescue PTCA does not appear to offer significant benefit beyond that of contemporary medical therapy after thrombolytic failure. PMID- 7484893 TI - Usefulness of combined two-dimensional echocardiography and transesophageal atrial stimulation early after acute myocardial infarction. AB - Transesophageal atrial stimulation (TRAS) was combined with 2-dimensional echocardiography in 69 consecutive patients on days 3 to 5 (mean 3.3) of their first, uncomplicated myocardial infarction, to determine if inducible remote asynergy (i.e., not directly adjacent to the infarcted area and supposedly related to another vascular territory) provides information regarding (1) extent of coronary artery disease, and (2) future ischemic events. Uncomplicated, adequate stress studies were performed in 59 of 69 patients (86%); all these patients had regional asynergy at rest. Remote asynergy at rest was present in 7 patients and during TRAS in 26 patients. Coronary angiography was performed within 2 to 3 weeks after the acute phase. Multivessel disease was present in 23 of these patients and absent in 3. Of the 33 patients without remote asynergy during TRAS, 5 had multivessel disease. Sensitivity of remote asynergy during TRAS for detecting multivessel CAD was 82%, specificity 90%, and predictive accuracy 86%. New ischemic events, defined as recurrent infarction, cardiac death, or revascularization within 12 to 18 months (mean 12.6) occurred in 24 patients (41%); remote asynergy during TRAS was present in 16 of these patients (67%). It is concluded that TRAS combined with 2-dimensional echocardiography can safely be performed in the early days of acute myocardial infarction; remote asynergy during TRAS reliably identifies patients with multivessel disease and future ischemic events. PMID- 7484894 TI - Prognostic value of predischarge exercise stress echocardiography after acute myocardial infarction. AB - A predischarge exercise test was performed by 70 patients 7 +/- 4 days (mean +/- SD) after acute myocardial infarction (AMI) to determine the short- and long-term prognostic value of predischarge exercise stress echocardiography (Ex-Echo) compared with exercise stress electrocardiography (Ex-ECG). Two-dimensional echocardiograms were obtained at rest and immediately after exercise; a wall motion score index was obtained both at rest and immediately after exercise. Results of the Ex-Echo were positive in 27 patients (39%), whereas those of Ex ECG were positive in 34 (49%). The wall motion index after exercise was lower in patients who died during follow-up (85 vs 98, p = 0.01) and in those with cardiac events, defined as death, nonfatal reinfarction, or revascularization (88 vs 98, p = 0.005). More patients with a positive Ex-Echo result had short-term cardiac events (within 2 weeks) than patients with a negative Ex-Echo (6 [22%] vs 2 [5%], p = 0.04). The same was true for long-term mortality (12 [44%] vs 3 [7%], p = 0.0002), reinfarctions (10 [37%] vs 4 [9%], p = 0.01), revascularization procedures (11 [41%] vs 7 [16%], p = 0.023), and cardiac events (22 [81%] vs 12 [28%], p < 0.0001). Survival time was shorter in patients with positive compared with negative Ex-Echo results (34% difference between groups, 95% confidence interval [CI] 10% to 58%, p = 0.002). The same applied for cumulative survival free from cardiac events (43%, p = 0.001, 95% CI 9% to 77%.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7484895 TI - Do gender-based differences in presentation and management influence predictors of hospitalization costs and length of stay after an acute myocardial infarction? AB - Previous studies have reported conflicting results on gender differences in the management of acute myocardial infarction (AMI) and have not evaluated hospital length of stay or costs. To determine gender-based differences in presentation, management, length of stay, costs, and prognosis after AMI, we studied 561 patients with AMI. Women were older, had systemic hypertension, diabetes mellitus, and a non-Q-wave AMI more frequently, whereas more men smoked cigarettes. Predictors of coronary angiography were: male gender (RR 1.9; 95% CI 1.2 to 3.1), chest pain at presentation (RR 1.8; 95% CI 1.0 to 3.3), recurrent angina (RR 4.1; 95% CI 2.5 to 6.8), admission via the emergency room (RR 0.2; 95% CI 0.1 to 0.3), and younger age. Gender did not predict mortality. Among presenting features, the predictors of length of stay were diabetes, prior coronary bypass and prior coronary angioplasty in men, and age alone in women. Pulmonary edema and need for coronary bypass during the hospital course were predictors of length of stay in men only. Among presenting features, predictors of cost were diabetes in men and congestive heart failure in women. Predictors of cost during hospitalization for men were pulmonary edema, coronary angiography, intraaortic balloon pump use, and coronary bypass; for women, they were peak levels of creatine kinase and coronary bypass. Thus, predictors of length of stay and hospitalization costs differ based on gender. Efforts at cost containment may need to be gender-specific. PMID- 7484896 TI - Influence of coronary angioplasty on the progression of coronary atherosclerosis. AB - This study examines the effect of coronary angioplasty on the progression and appearance of new disease in sites of the coronary tree that were not dilated by the balloon. We examined 355 pairs of coronary angiograms from 252 patients. The study consisted of consecutive patients who were referred for catheterization > 1 month after successful angioplasty. Progression/regression and the appearance of new narrowings at sites not dilated by angioplasty were determined. The life table method was used to determine outcome, and any event (progression, regression, and new narrowing) was analyzed according to the time of occurrence. The angioplasty artery was compared with the non-angioplasty artery and the effect of restenosis was determined by comparing arteries with and without restenosis. Progression/regression rates were not significantly different in angioplasty and non-angioplasty arteries. More new narrowings were identified in the angioplasty artery (p < 0.01). With regard to narrowings located in the angioplasty artery, progression was more common, regression less common, and the appearance of new narrowings more common in arteries with restenosis than in non angioplasty arteries or arteries without restenosis. We believe that mechanical trauma to the artery during angioplasty could accelerate disease progression and the appearance of new narrowings in angioplasty arteries, whereas normalization of flow rate and pattern, especially in arteries without restenosis, attenuates the rate of progression and the appearance of new narrowings in these arteries. The final outcome depends on the balance between these factors. PMID- 7484897 TI - Time course and mechanism of early luminal diameter loss after percutaneous transluminal coronary angioplasty. AB - To assess the time course and mechanism of early minimal luminal diameter (MLD) loss, serial angiographic observations were performed. Seventy-four patients (with 74 severe narrowings [ > or = 70%]) with acute ischemic syndromes who had an early loss in MLD of > 0.3 mm at 24 hours after percutaneous transluminal coronary angioplasty (PTCA) also underwent 1 hour post-PTCA angiography. In 12 consecutive patients with early loss 1 hour after PTCA, angioscopy was also performed to assess the mechanism of early loss. The percent diameter stenosis for the 74 lesions was 16.8 +/- 8.4% immediately after PTCA, 35.1 +/- 14.2% 1 hour after PTCA (p < 0.002 vs immediately after), and 41.4 +/- 13.2% at 24 hours (p < 0.10 vs 1 hour after). The MLD also showed similar differences: 2.6 +/- 0.3 mm immediately after to 2.0 +/- 0.4 mm 1 hour after(p < 0.002) to 1.8 +/- 0.4 mm 24 hours after PTCA (p < 0.10 vs 1 hour). In 60 patients (81%), the > 0.3 mm loss was detected 1 hour after PTCA. These 60 patients had no further decreases in MLD at 24 hours (1.9 +/- 0.4 vs 1.8 +/- 0.4 mm at 1 and 24 hours, respectively, p = NS). Adequate angioscopic images available in 11 patients showed that red thrombus was present in 1, minor or multiple dissection in 5, and neither thrombus nor dissection in 5 other patients (consistent with early wall recoil). Thus, in narrowings demonstrating early loss in MLD at 24 hours, 81% showed that the early loss occurred within 1 hour after PTCA. Early loss is not related to thrombus but usually to dissection or recoil. PMID- 7484898 TI - Coronary lumen at six-month follow-up of a new radiopaque Cordis tantalum stent using quantitative angiography and intracoronary ultrasound. AB - To determine the reliability of geometric (edge-detection) quantitative coronary angiographic analysis (QCA) of restenosis within a new Cordis tantalum stent, QCA and intracoronary ultrasound (ICUS) measurements were compared in both an experimental restenosis model and in the clinical follow-up of patients. In the experimental series, Plexiglas phantom vessels with concentric stenosis channels ranging from 0.75 to 3.0 mm in diameter and with a reference diameter of 3.0 mm were imaged both before and after their insertion in tantalum stents. In the clinical series, the agreement of QCA and ICUS measurements were studied in 23 patients who had undergone coronary implantation of the new tantalum stent and in 23 patients who had undergone balloon angioplasty 6 months previously. The reliability of QCA declined in the presence of the radiopaque stent (accuracy of QCA decreased from -0.07 to -0.12 mm), whereas the reliability of lumen measurements by ICUS was independent of the presence of the radiopaque stent ( 0.12 and -0.13 mm). Without the stent, the average minimal luminal diameter (MLD) obtained by QCA of the 1.00 mm Plexiglas vessel was 1.00 +/- 0.01 mm, and the 3.00 mm reference vessel diameter was 2.81 +/- 0.05 mm, providing a 64 +/- 1% diameter stenosis. After introduction of the stent, the average MLD and reference vessel diameter were 0.99 +/- 0.06 and 3.36 +/- 0.17 mm, respectively, providing a diameter stenosis of 71 +/- 2%. ICUS measurements (2.77 mm) of the reference vessel diameter (3.00 mm) were unaffected by the presence of the stent.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7484899 TI - Usefulness of a prototype directional catheter for excimer laser coronary angioplasty in narrowings unfavorable for conventional excimer or balloon angioplasty. AB - We report clinical and angiographic results in 53 patients with 57 significant coronary or saphenous vein graft narrowings treated with directional excimer laser angioplasty. The target vessels were the left main (1%), anterior descending (32%), circumflex (19%), right coronary artery (39%), and vein grafts (9%). Lesions were morphologic class B1 (18%), B2 (79%), or C (3%), with 40 de novo and 17 restenotic lesions. Adjunctive balloon angioplasty was used in 53 lesions (93%). Mean pre- and postprocedural minimal lumen diameters were 0.6 +/- 0.3 and 1.9 +/- 0.7 mm (p < 0.001), corresponding to a mean diameter stenosis of 72 +/- 20% and 27 +/- 16%. Procedural success rate was 91%. Cumulative risk of death, Q-wave myocardial infarction, or emergency bypass operation was 9% (5 patients). Of patients who had a successful laser procedure, 28 (60%) with 30 lesions underwent angiographic follow-up at 6 +/- 3 months after the procedure. Restenosis rates (> 50% diameter restenosis or acute gain loss) were 37% and 23%, respectively. Four patients underwent bypass, 3 angioplasty, and 1 patient died from cancer. This study demonstrates the feasibility of directional application of laser energy to selected unfavorable narrowings for conventional excimer laser or balloon angioplasty. Further evaluation of this device using the now standard saline infusion technique is necessary to establish its ultimate role as a primary interventional device. PMID- 7484900 TI - Comparison of the diagnostic accuracy of different methods of measurement of heart rate-adjusted ST-segment depression during exercise testing for identification of coronary artery disease. AB - Within a population of 160 consecutive symptomatic patients who all had undergone catheterization (80 with > or = 1 stenosis > or = 50%), we compared the accuracy of different computerized measurements of the exercise-induced changes in ST segment: (1) the standard criterion (> or = 0.1 mV flat/downsloping ST depression or > or = 0.15 mV upsloping depression, both 60 ms after the J point); (2) heart rate (HR)-adjusted ST-segment depression (ST/HR index measured at 0, 20, 40, 60, and 80 ms from the J point); (3) the HR-adjusted ST integral (ST/HR integral measured from 0 to 40 ms and from 40 to 80 ms after the J point). None of the ST/HR indexes or integrals were found to have a significantly greater sensitivity than the standard criterion. On the contrary, all ST/HR indexes and integrals showed a higher specificity (0.78 to 0.89) than did the standard criterion (0.65); moreover, the earlier the measurement within the repolarization phase, the better the overall accuracy: 0.71 for the standard criterion, 0.83 (p < 0.001), 0.80 (p < 0.01), 0.78 (p < 0.02), 0.78 (p < 0.02), 0.74 (p = NS) for the ST/HR indexes at 0, 20, 40, 60, and 80 ms, respectively; 0.81 (p < 0.001) and 0.78 (p < 0.02) for the ST/HR integrals calculated from 0 to 40 and from 40 to 80 ms, respectively. Consistently, the receiver-operating characteristic curve areas of ST/HR at 0, 20, and 40 ms were greater than those of ST/HR at both 60 and 80 ms. These findings are divergent from some other results given in published reports. We conclude that the accuracy of all exercise criteria is influenced by the population analyzed: our patients were representative of those currently seen by clinicians. PMID- 7484901 TI - Fasting hyperinsulinism, insulin resistance syndrome, and coronary artery disease in men and women. AB - A large segment of the population gradually develops insulin resistance, and the related metabolic syndrome is one of the most frequent causes of atherosclerosis. Searching for a practical indicator of insulin resistance, we studied the correlations between fasting serum insulin level, the general manifestations of insulin resistance syndrome, and various aspects of coronary artery disease in 797 men and 322 women. After we classified patients according to the quartiles of serum insulin level, we noted in the top quartile the presence of practically all manifestations of insulin resistance syndrome in persons of both sexes (e.g., increased waist/hip ratio, body mass index, glucose, uric acid, triglycerides, apolipoprotein B and decreased high-density lipoprotein cholesterol levels as well as apolipoprotein A-I/B ratios, and so forth). We also noted a higher prevalence of hypertension, diabetes mellitus, and type IV hyperlipidemia. Significantly more women in the fourth than in the first quartile had angiographically documented significant stenosis of the coronary arteries (p = 0.0016, odds ratio 2.9, 95% confidence interval 1.5 to 5.6) and previous myocardial infarction (p = 0.0297, odds ratio 2.1, 95% confidence interval 1.1 to 4.1). Men in both the first and the fourth quartile had a more disturbed lipid profile and a higher prevalence of significant stenoses of coronary arteries and/or previous myocardial infarction than women; there was a tendency toward a lower prevalence of alcohol consumption (p = 0.0503), a higher prevalence of gout (p = 0.0634), and previous myocardial infarction (p = 0.0791) in men in the fourth than in the first quartile.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7484902 TI - Evidence that endothelial dysfunction in patients with hypercholesterolemia is not due to increased extracellular nitric oxide breakdown by superoxide anions. AB - Patients with hypercholesterolemia have impaired endothelium-dependent vasodilation due to decreased nitric oxide activity. The present study aimed to determine whether this form of endothelial dysfunction is related to enhanced extracellular breakdown of nitric oxide by superoxide anions. To this end, the vascular responses to acetylcholine (an endothelium-dependent vasodilator) and sodium nitroprusside (a direct smooth muscle dilator) were studied before and after combined administration of copper-zinc superoxide dismutase (a scavenger of superoxide anions with poor intracellular penetrance; 6,000 U/min) in 20 normal controls (11 men and 9 women, age 50 +/- 6 years) and in 20 hypercholesterolemic patients (10 men and 10 women, age 49 +/- 9 years). Drugs were infused into the brachial artery and the response of the forearm vasculature was measured by plethysmography. The vasodilator response to acetylcholine was significantly blunted in hypercholesterolemic patients compared with normal controls (maximal flow 8.8 +/- 2 vs 12.7 +/- 3 ml/min/100 ml, respectively; p < 0.03); however, no difference was observed in the response to sodium nitroprusside (9.7 +/- 2 and 9.5 +/- 3 ml/min/100 ml). In normal controls, the infusion of superoxide dismutase did not significantly modify the response to acetylcholine (maximal flow 12.7 +/- 3 vs 12.1 +/- 3 ml/min/100 ml before and after superoxide dismutase, respectively). Similarly, in hypercholesterolemic patients, the infusion of superoxide dismutase did not alter the response to acetylcholine (maximal flow 8.8 +/- 2 and 8.9 +/- 2 ml/min/100 ml). A subset of 19 subjects (8 normal and 11 patients) received a 60-minute infusion of superoxide dismutase at 24,000 U/min without alteration in their response to acetylcholine.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7484903 TI - Adequacy of implantable cardioverter-defibrillator lead placement for tachyarrhythmia detection by sinus rhythm electrogram amplitude. AB - This study examines whether the current clinical practice of using a 5 mV minimum amplitude during normal sinus rhythm (NSR) ensures adequate detection during subsequent episodes of ventricular fibrillation (VF) at the time of the implantable cardioverter-defibrillator (ICD) threshold testing. Risk of nondetection occurs with ICDs when a substantial portion of the individual cardiac events on an electrogram goes undetected. Detection risk was assessed by 2 methods: percentage of missed cardiac events (incidence of signal dropout), and mean electrogram amplitude. During ICD implantation and testing in 27 patients utilizing 41 lead positions, 135 episodes of VF were induced and analyzed. On 64 occasions, the countershock was not successful in achieving cardioversion, and the continuing electrical activity was analyzed as a separate group of postshock waveforms. Thresholds of 1 and 2 mV were applied to each individual cardiac depolarization in a VF episode. Significant risk of nondetection was assumed when > or = 10% of individual events displayed dropout. Underdetection by signal dropout occurred in 11 of 135 preshock arrhythmia signals (8.1%) from 3 patients at a 2 mV threshold, and in 6 of 135 signals (4.4%) at a 1 mV threshold. A mean NSR amplitude > or = 5 mV was associated with significantly lower risk of nondetection during subsequent VF episodes at both 1 and 2 mV thresholds (largest p < 0.001). Similar results were observed in analysis of postshock arrhythmia signals. Further examination of signal dropout and linear regression criteria suggest that in order to eliminate the possibility of nondetection at a 1 mV threshold, minimum NSR amplitudes of 8.5 and 10.0 mV, respectively, are required. PMID- 7484904 TI - Effect of donor-recipient size mismatch on left ventricular remodeling after pediatric orthotopic heart transplantation. AB - Left ventricular (LV) hypertrophy has been reported after orthotopic heart transplantation. This study was designed to determine the pattern of LV remodeling in the first year after pediatric orthotopic heart transplantation and to elucidate the mechanism responsible for changes in LV dimensions. Serial echocardiograms of 20 children who underwent cardiac transplantation were analyzed off-line, and the following LV parameters were measured and indexed to body surface area (BSA): short-axis diameters, posterior wall thickness, length, mass, and volume in systole and diastole. Mass/volume and short-axis diameter/length ratios and ejection fraction were calculated. In 5 patients, the donor's echocardiogram was also available for analysis. The patient's systemic blood pressure at the time of the echocardiogram, ischemic time of the donor heart, number of rejection episodes, biopsy scores, and body size of the donor and patient were recorded. Patients were assigned to 2 groups based on their donor-recipient weight ratio:group 1, < or = 1.2 (n = 9); and group 2, > 1.2 (n = 11). In group 1, LV mass index remained within normal limits throughout the study period. In group 2, mass index was significantly increased 2 weeks after transplantation (72 +/- 24 vs 133 +/- 37 g/BSA1.5, p = 0.0008). LV volume, geometry, ejection fraction, systemic blood pressure, and number of rejection episodes did not differ significantly between groups. The excess LV mass index in group 2 regressed significantly during the first year after transplantation from 133 +/- 37 to 93 +/- 17 g/BSA1.5 (p < 0.005).(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7484905 TI - Early abnormalities of cardiac function in non-insulin-dependent diabetes mellitus and impaired glucose tolerance. AB - The aim of this study was to evaluate the role of diabetes and minor abnormalities of glucose homeostasis, such as impaired glucose tolerance, as determinants of cardiac function and structure in a working population. We studied a population-based sample of 64 telephone company employees (both sexes, mean age 58 years): 25 with normoglycemia, 15 with impaired glucose tolerance, and 24 with non-insulin-dependent diabetes mellitus (NIDDM) diagnosed by oral glucose tolerance test according to the recommendations of the World Health Organization. Subjects with myocardial ischemia were excluded. Left ventricular end-systolic dimension, indexed to body surface area, was greater in those with NIDDM (p < 0.05) and in those with impaired glucose tolerance (p < 0.05) with respect to normoglycemic persons. The ratio of the peak early diastolic velocity wave to the late diastolic wave was lower in those with NIDDM (p < 0.05) and in those with impaired glucose tolerance (p < 0.05) than in participants with normoglycemia. Body mass index and blood pressure were similar in the 3 groups. These results clearly indicate that early abnormalities of cardiac structure and function are observed not only in patients with NIDDM, but also in those with impaired glucose tolerance, independent of the confounding role of myocardial ischemia, body weight, and blood pressure. PMID- 7484906 TI - The way of the future redux. PMID- 7484907 TI - How good is echocardiography at assessing myocardial viability? PMID- 7484908 TI - Combination of lovastatin, enalapril, and colchicine does not prevent restenosis after percutaneous transluminal coronary angioplasty. PMID- 7484909 TI - Effects of vitamin E on endothelial function in men after myocardial infarction. AB - This study showed that endothelial dysfunction is present in men 3 to 6 months after myocardial infarction, but was unable to show any improvement in endothelial function after 3 months of therapy with vitamin E 800 IU/day. Further studies are necessary to determine whether higher doses or a longer course of vitamin E, or whether other antioxidant agents with or without lipid-modifying activity, would improve endothelial function. PMID- 7484910 TI - Echocardiographic findings associated with atrial fibrillation in 1,699 patients aged > 60 years. PMID- 7484911 TI - Utility of QT dispersion combined with the signal-averaged electrocardiogram in detecting patients susceptible to ventricular tachyarrhythmia. AB - A simple algorithm that combines QT dispersion with the signal-averaged electrocardiogram QRS duration provides an extremely sensitive method for predicting spontaneous or inducible ventricular tachyarrhythmias. This new algorithm may prove useful in determining which patients are at risk for ventricular tachyarrhythmia. PMID- 7484912 TI - Relation of duration of morbid obesity to left ventricular mass, systolic function, and diastolic filling, and effect of weight loss. AB - Longer duration of morbid obesity is associated with higher LV mass, poorer LV systolic function, and greater impairment of LV diastolic filling. Weight loss induced decreases in LV mass and improvements in LV systolic function and diastolic filling are due in part to favorable alterations in LV loading conditions. PMID- 7484914 TI - Heart rate variability in pheochromocytoma. AB - The results of our study show that parasympathetic tone was higher in patients with pheochromocytoma than in patients with primary hypertension. An unusual spectral form of vagal activity was seen during excessive beta-adrenergic stimulation, while persistent hypertension with an excessive alpha-adrenergic stimulation was responsible for low cardiac sympathetic tone, probably due to desensitization of beta-adrenergic receptors. PMID- 7484913 TI - Effect of weight loss on left ventricular diastolic filling in morbid obesity. PMID- 7484915 TI - Prospective evaluation of complications associated with percutaneous intraaortic balloon counterpulsation. AB - In summary, current, prospective experience with 691 consecutive patients revealed that female gender, peripheral vascular disease, cardiac index, and diabetes were the only independent predictors of risk after percutaneous IAB insertion. Neither sheathless insertion nor smaller IAB catheter size was associated with a lower complication rate. There may be a threshold of catheter size associated with a dramatic reduction in complications. If that exists, it has not yet been reached. PMID- 7484916 TI - Electrocardiographic abnormalities in patients with acute aortic dissection. AB - In conclusion, acute electrocardiographic changes were not uncommon in patients with acute aortic dissection. It was suggested that acute electrocardiographic changes in aortic dissection resulted from at least 1 of the following 3 mechanisms: (1) involvement of the ostium of the coronary artery; (2) shock state, especially tamponade; and (3) preexisting coronary artery disease. We believe that physicians should be aware of the possibility of acute aortic dissection not only in patients with chest pain with a normal electrocardiogram, but also in those with electrocardiographic changes. PMID- 7484917 TI - Advances in human energy metabolism: balancing energy requirements and energy intake. Proceedings of a symposium. Beltsville, Maryland, May 8-11, 1994. PMID- 7484918 TI - The calorie: myth, measurement, and reality. AB - Few dietary components are surrounded by more misinformation and myths than the calorie. This confusion can be attributed in part to a lack of accurate and practical methods for assessing energy intake and thus requirements in humans over periods extending beyond several days. The availability of modern respiratory-chamber indirect calorimetry systems and results from human studies with doubly labeled water are now helping to clarify uncertainties surrounding energy requirements. We describe studies of patients with endogenous obesity as an example of how these research methods are resolving long-standing questions regarding energy requirements. The results of these investigations reveal some of the flaws in estimating energy requirements by self-report methods. Advances in accurately measuring energy expenditure are making important contributions to the study of human energy requirements and are providing new and important research opportunities. PMID- 7484919 TI - Energy expenditure measurements in relation to energy requirements. AB - Long-term good health for weight-stable adults requires balance between energy intake (EI) and energy expenditure (EE). Both EI and EE measurements can be used to estimate energy requirement. Results from studies conducted at Beltsville Human Nutrition Research Center in which two or more methods were used to measure EE are compared to determine relative differences in measurement techniques. Comparison of EI estimated from dietary intake records (7 d minimum) with EI measured in 12 controlled feeding studies (45 d minimum) in 266 subjects indicated that diet records underestimate energy requirement by 18%. Comparison of EE measured in a room calorimeter and by doubly labeled water (2H(2)18O) in nine subjects showed no significant difference between methods within the calorimeter environment (1.6 +/- 2.6%) but free-living EE measured by 2H(2)18O was greater (13.2 +/- 7.1%) due to physical activity. Comparison of metabolizable energy (ME) intake, 2H(2)18O, and direct or indirect calorimetry in four subjects indicated no difference between ME and EE measured by 2H(2)18O (-1.0 +/- 1.3%) or between direct and indirect calorimetry (0.6 +/- 0.9%); however, free-living EE measured by 2H(2)18O was greater than the value calculated by calorimetry (15.3 +/- 5.0%). Each method has associated limitations that include availability, accuracy, precision, and cost. Appropriate application and interpretation of results for all methods are essential. PMID- 7484920 TI - Energy requirements from infancy to adulthood. AB - To investigate how age and body composition affect energy requirements, the sedentary daily expenditure (SDE) and basal metabolic rate (BMR) of 101 infants, 82 girls, and 27 adults were measured. Energy expenditure was scaled for differences in body size to test the effects of age and body fatness. A power function was superior to linear models. For all subjects, WT0.63 (where WT is weight) or FFM0.63 (where FFM is fat-free mass) explained 94% of the variability in BMR, and WT0.70 or FFM0.70 explained 97% of the variability in SDE. The effects of height and fat mass (kg or % body wt) on BMR and SDE scaled for weight or fat-free mass were age dependent. Best-fitted exponents relating BMR or SDE to body size differed between children (0.40-0.52) and infants (1.04-1.30) (P = 0.001). Human energy requirements from infancy to adulthood appear to be a power, not a linear, function of body weight and composition. PMID- 7484921 TI - Influence of age on energy requirements. AB - The effects of aging on energy requirements and energy expenditure were investigated in 35 healthy young men (mean +/- SE age, 22.7 +/- 0.6 y) and elderly men (68.0 +/- 1.5 y). Over a 10-d metabolic balance study, measurements were made of metabolizable energy intake for weight maintenance, total energy expenditure, resting energy expenditure, and thermic effect of feeding. Values for both metabolizable energy intake and total energy expenditure were significantly higher than the current recommended dietary allowance for energy in both age groups (P < 0.01). Total energy expenditures were 14.48 +/- 0.65 and 11.26 +/- 5.40 MJ/d in young and elderly men, respectively. The difference in total energy expenditure between the groups was accounted for by a significant decrease in all the major components of expenditure. These results suggest that the current recommended dietary allowances for energy may underestimate the usual energy needs of healthy adult men. PMID- 7484922 TI - Physical activity and energy requirements. AB - Differences in physical activity represent the largest source of variability in energy requirements, both within and between individuals. Chronic changes in physical activity can produce chronic changes in energy requirements that, if not compensated for, can lead to changes in the level at which body weight and body composition are maintained over time. We present a model that can serve as a framework for understanding how energy and macronutrient balance are maintained in steady state conditions, and for illustrating the potential effect of a change in physical activity on these steady state conditions and on energy requirements. According to the model, a chronic change in physical activity forces changes in other aspects of energy and substrate utilization so that a steady state condition can be reestablished. The net effects of the change in physical activity on energy requirements will depend on how and over what period of time these new steady states are reached. Although we cannot at present predict, for any individual, the precise effect of changes in physical activity on energy requirements and on body weight and composition, our model provides a framework for further study of the factors that can influence energy requirements. PMID- 7484923 TI - Ethnicity and energy stores. AB - The primary storage form of energy within humans is fat, which accumulates in adipose tissue including the subcutaneous, omental, mesenteric, retroperitoneal, and mammary depots. Although it has been known for some time that the size of these depots varies with sex, age, and physiologic state, it has only recently been suggested that adipose tissue partitioning, and therefore energy storage, may vary among ethnic groups. Indicators of ethnicity include race, place of birth, and culture and traditions. The literature describing energy storage in North American Indians, African Americans, Asian Americans, Mexican Americans, and Pacific Islanders is summarized and data are presented from studies comparing African American and Caucasian (Americans of Northern European descent) obese women. It is proposed that, for the purpose of research, physiologic characteristics and not ethnicity should be the basic factors used to recruit human study volunteers until we obtain mechanisms to discriminate genotype and to relate phenotype to energy storage. PMID- 7484924 TI - Total energy intake of the US population: the third National Health and Nutrition Examination Survey, 1988-1991. AB - The third National Health and Nutrition Examination Survey (NHANES III) was conducted to assess the health and nutritional status of the US population. As part of the nutritional status assessment, reliable 24-h dietary recalls were collected for 14,801 examined persons. Mean (+/- SEM) energy intakes are reported for persons aged > or = 2 mo by age, sex, and race-ethnicity. Males had higher mean energy intakes than did females. Energy intakes peaked during late adolescence and young adulthood and declined thereafter. Energy intake patterns were similar among non-Hispanic whites, non-Hispanic blacks, and Mexican Americans. Underreporting was addressed by computing a ratio of energy intake (EI) to estimated basal metabolic rate (BMRest). This ratio (EI:BMRest) was 1.47 for adult males and 1.26 for nonpregnant adult females. Overweight adults had a lower mean EI:BMRest (1.09 in females and 1.28 in males). Underreporting in food consumption surveys remains problematic among females and overweight persons. PMID- 7484925 TI - Energy intake and sensory properties of food. AB - Sugars and fat play a unique role in the human diet. The selective choice of sugars and fat as chief energy sources seems to be influenced less by the body's energy needs than by the sensory appeal of sweet and fat-rich foods. This appeal typically holds not only across individuals but across cultures. Although many behavioral, social, and cultural factors play major roles in diet selection, people respond primarily to the sensory qualities of food. Some clinical studies have reported that individual food choices and therefore the macronutrient composition of the diet are influenced directly by the central nervous system. However, broader population-based studies point to the central role of taste in determining food selection. Survey studies have shown that the global consumption of sugars and fat is further determined by such variables as income, socioeconomic status, and the availability of sugars and fat in the food supply. Nutrition intervention strategies aimed at promoting dietary change in communities ought therefore to consider not only physical health, but also the sensory pleasure response, and a wide range of demographic, economic, and sociocultural variables. PMID- 7484926 TI - Fat, carbohydrate, and the regulation of energy intake. AB - High-fat foods are readily overeaten. This could be because fat increases the palatability and energy density of foods or because fat has a low satiety value compared with carbohydrate. In several studies examining the effects of preloads that varied in fat and carbohydrate content on subsequent food intake, there was little difference in the effects of fat and carbohydrate on satiety. A dose response preloading paradigm gives the most sensitive index of satiety. In one such test it was found that the effects of yogurts varying in fat or carbohydrate content did not differ in normal-weight, unrestrained men. However, in obese individuals or those concerned with body weight, fat in the yogurts was less effective in reducing subsequent food intake than carbohydrate. More studies are required to determine whether this small, but significant, insensitivity to energy from fat affects the regulation of food intake and body weight. PMID- 7484927 TI - Control of energy intake by energy metabolism. AB - Eating behavior is controlled by signals that are generated in the postabsorptive metabolism of energy-yielding substrates. Recent work indicates that an event common to the metabolism of glucose and fat provides such a signal. Evidence suggests that eating behavior is triggered by a signal that is tied to hepatic ATP concentrations and is carried from the liver to brain via afferents in the vagus nerve. This metabolic control of eating behavior may link mechanisms of energy storage and expenditure to energy intake. Changes in energy intake associated with alterations in energy expenditure can be viewed as a response to a shift in the partitioning of fuels, which affects the oxidative metabolic pathways that generate the signal controlling eating behavior. PMID- 7484928 TI - Alcohol and energy intake. AB - One hundred years of research about the metabolism of alcohol have provided many details, but some general aspects of the physiologic value of alcohol remain uncertain or inadequately proven. Results from epidemiologic studies appear to be in conflict with interpretations based on results from indirect calorimetric studies. The apparent inability of body mass index to be maintained in women when alcohol is consumed with food may indicate impaired metabolic processes that need to be better understood. Current evidence on the effects of alcohol are summarized to identify experimental approaches that may provide information needed to resolve the current contradictions. PMID- 7484929 TI - Body composition, respiratory quotient, and weight maintenance. AB - For weight maintenance, the fuel mix oxidized must match the nutrient distribution in the diet. The composition of the fuel mix oxidized and the respiratory quotient are influenced by circulating substrate and hormone concentrations, which, among other things, reflect the degree of replenishment of the body's fuel reserves. Carbohydrate and protein balances are maintained without obvious changes in the body's glycogen stores and protein pools. One of the most readily quantifiable measures by which to judge the effect of various inherited and circumstantial factors on energy metabolism and body composition is the percentage body fat for which weight maintenance occurs. The amount of body fat for which the steady state of weight maintenance tends to become established is therefore a key fact on which an understanding of body weight regulation should be based, rather than rates of energy turnover or past circumstances that may have led to fat accumulation, as often believed. PMID- 7484930 TI - Sympathetic nervous system response to intake. AB - Meals increase the activity of the sympathetic nervous system (SNS) above the activity level observed in the postabsorptive state. Pharmacologic blockade of the thermogenic effect of norepinephrine, the primary neurotransmitter of the SNS, suggests that this response accounts for approximately 20% of the thermic response to food. Energy expenditure increases after several days of sustained overeating. There is no convincing evidence that this response is mediated by increased SNS activity in humans, as it is in some animals. Energy expenditure decreases within a few days of energy deprivation. When sodium balance is maintained, energy deprivation also reduces SNS activity. The reduced SNS activity may contribute to the decreased energy expenditure, but cannot explain it completely. The contribution of the SNS to total energy expenditure is small in normal subjects consuming a weight-maintenance diet, probably < 5%. A few studies have suggested that SNS activity is reduced in obese humans, suggesting that low energy expenditure associated with reduced SNS activity may contribute to obesity. However, a critical evaluation of the available evidence indicates that this hypothesis is untenable. PMID- 7484931 TI - New model for the regulation of energy balance and adiposity by the central nervous system. AB - We describe a new model for adiposity regulation in which two distinct classes of peripheral afferent signals modulate neuronal pathways in the brain that control meal initiation, meal termination, and the autonomic outflow influencing the fate of ingested energy. These brain pathways, termed central-effector pathways for the control of energy balance, respond to 1) short-term, situational-, and meal related signals that are crucial to the size and timing of individual meals, but that are not components of the system serving to regulate adipose stores, and 2) long-term, adiposity-related signals that participate in the negative feedback control of fat stores. Long-term signals, such as the pancreatic hormone insulin, are secreted into the circulation in proportion to energy balance and adipose mass. These signals enter the brain where they influence central-effector pathways, in part by changing the sensitivity of these pathways to short-term signals. Through this mechanism, the central nervous system response to short term signals is adjusted in proportion to changes in body adiposity, resulting in compensatory changes in food intake and energy expenditure that collectively favor the long-term stability of fat stores. This model provides a comprehensive framework for experimental design and data interpretation in the study of body adiposity regulation. PMID- 7484932 TI - Metabolizable energy of macronutrients. AB - I describe recent advances in assessing the amount of energy available from diets of varied composition. Empirical models of food energy prediction have grown in preference to factorial models over the past 20 y and knowledge of the quantitative aspects of energy salvage in the colon have modified our thoughts on how to best calculate food energy values. In contrast, food regulatory practices have been limited mostly to changes in the way in which carbohydrate is defined or measured and there has been little change in the basic approach to energy evaluation or the energy conversion factors used with food components. The empirical systems have advantages over factorial models of food energy assessment and it is proposed that such empirical systems should be used in food regulations, food tables and databases, and in nutritional studies in which knowledge of metabolizable energy intake is desirable but cannot be determined directly. PMID- 7484933 TI - Nutrition labeling: energy values of foods and fat substitutes. AB - The Federal Food, Drug, and Cosmetic Act requires the listing of all food ingredients on the label. However, until this year, nutrition labeling for most foods was not required. The Nutrition Labeling and Education Act, passed by the US Congress in November 1990, requires nutrition labeling (on the package or at the site of purchase) of virtually all foods packaged after May 8, 1994. (A law was passed by Congress and signed by the President of the United States on May 26, 1994, that delayed applicability of the Nutrition Labeling and Education Act until after August 8, 1994, for certain products whose labels were printed before May 8, 1994, and for which there was supporting documentation that after August 8, 1994, the product labels would be in compliance.) Prominent among the nutrition information required is labeling of the total energy content of food and the energy content derived from fat. In Title 21, Code of Federal Regulations, five methods are specified for determining the energy content of foods. The US Food and Drug Administration expects also to include specific food factors (digestibility coefficients) in food additive and GRAS (generally recognized as safe) listings for calculating the energy values of novel foods and ingredients. PMID- 7484935 TI - Food sampling strategies for energy intake estimates. AB - Current interest in the relation of diet to the maintenance of good health has stimulated the demand for food composition data. Values for energy, nutrients, and other food-based components are required to calculate intake, to provide an adequate food supply, to formulate and label new products, and to facilitate trade. The US Department of Agriculture National Nutrient Databank serves as the foundation for food composition databases used by the food industry, government agencies, and academia for these activities. To be accurate, a specific estimate in a database must be statistically representative of the universe of all values for a component in a food. Statistically based sampling strategies must be developed to identify and select appropriate units of specific foods for analysis. The determination of energy values for foods is a unique challenge for food sampling because energy values are calculated from values for total fat, protein, and carbohydrate fractions. To establish priorities for sampling, food consumption data can be used to rank the primary contributors of energy for the population of interest. Demographic and marketing data for foods can be used to identify specific products and locations for sampling. Users of food composition data should question the quality and representativeness of estimates for energy, including estimates for total fat, protein, and carbohydrate to be used for their applications. PMID- 7484934 TI - Nutrient Databank perspectives on energy. AB - The Atwater system of food-specific factors continues to be used for calculating energy in the Nutrient Databank. In 1983 the US Department of Agriculture requested that the Life Sciences Research Office (LSRO) of the Federation of American Societies for Experimental Biology reassess the system's accuracy and utility. An LSRO panel of experts reported that there was no evidence that a substantial problem exists. Advances in food technology and changes in analytical methods in the past decade, however, are causing some concern for the reporting of energy in the databank. Some perspectives on these and other issues are discussed. PMID- 7484936 TI - Contribution of fiber and resistant starch to metabolizable energy. AB - Recommendations made to increase complex carbohydrate and fiber intake in the United States may result in energy loss from increased fecal losses of starch, protein, and fat. The type of fiber or starch and the amount consumed affect digestion of the carbohydrate and interactions with other nutrients. On average, 8.4 kJ (2 kcal) digestible energy/g is available from up to 70 g poorly digested carbohydrate/d. Undigested fiber and starch are important substrates for colon bacteria and are fermented to short-chain fatty acids, hydrogen, and methane. Hydrogen and methane excreted through the lungs have been used as indicators of colonic fermentation. Fermentation appears to contribute significant digestible energy when > 20 g poorly digested carbohydrate/d is consumed. PMID- 7484937 TI - Sugar substitutes: their energy values, bulk characteristics, and potential health benefits. AB - Restriction of dietary energy extends life and reduces incidences of disease in animals. These benefits would likely extend to humans. However, diet restriction in animals imposes reductions of 30-50% in food intake, which is probably unacceptable to humans. Low-energy sweeteners used in beverages offer minor reductions in energy intake. However, they lack the bulk required for baked goods and other sugar-rich foods. Full-bulk sweeteners providing about one-half the energy of sugar are under development for such uses. Laxation limits their acceptable dose. Even within such limitations, they can help achieve the health benefits for humans indicated by diet restriction. D-Tagatose, a new candidate sweetener, is nearly as sweet as sucrose and has the bulk of sucrose, yet provides zero available energy. We discuss its potential contribution to human diet restriction along with its specific effect in delaying the aging effects of glycosylation. PMID- 7484938 TI - Methods available to estimate the energy values of sugar alcohols. AB - There is increased interest in the use of sugar alcohols as substitutes for sucrose in various food products. Part of this interest is derived from studies suggesting that sugar alcohols may have lower energy values because of the way they are metabolized. Contributing to the complexity is the fact that not all sugar alcohols are similarly metabolized. Indirect and direct methods used to assess the energy value of sugar alcohols have often yielded conflicting data. Energy values obtained using mathematical models have been adopted by some countries to account for metabolic processes associated with sugar alcohol digestion and absorption. I focus on two sugar alcohols, sorbitol and maltitol, and describe various methods that have been used to assess their energy value. PMID- 7484939 TI - Nutrient substitutes and their energy values in fat substitutes and replacers. AB - In recent years, new and redesigned ingredients have been introduced as fat substitutes and replacers. A fat replacer is an ingredient that replaces some or all of the functions of fat and may or may not provide nutritional value. A fat substitute is an ingredient that replaces all the functions of fat without any energy contribution. Currently available fat replacers are fat mimics (also called mimetics) or fat analogs. Fat mimics are based on carbohydrate, protein, and/or fat and have energy values from 0 to 38 kJ/g (0-9 kcal/g). Fat analogs have the characteristics of fat but have less energy. Because no one ingredient replaces all the functions of fat, most reduced-fat foods are formulated with a combination of ingredients and processes that affect both fat and energy content. Each formula must be evaluated for its total energy value; additionally, once acceptability and long-term use patterns of these reformulated products are established, their effect on energy intake must also be evaluated. PMID- 7484940 TI - Clinical and economic performance criteria for intraarterial and extraarterial blood gas monitors, with comparison with in vitro testing. AB - The frequency with which blood gas measurements should be obtained is controversial because laboratory-based blood gas analyzers impose blood loss, delay, and cost on each measurement. Point-of-care analyzers eliminate the delay factor, but blood loss and cost factors remain. Optode microsensing can be adopted readily to in vivo monitoring devices that can provide continuous or on demand blood gas values that do not result in blood loss or added cost to each measurement. This article critically reviews the advantages and limitations of optode-based blood gas monitors based on published data. In addition, a basis for considering the economic effects of extralaboratory blood gas measurement devices (point-of-care analyzers and optode-based monitors) is presented. PMID- 7484941 TI - Economic considerations of point-of-care testing. AB - Point-of-care testing is a rapidly expanding trend. This review discusses some of the issues driving the development of this new approach to laboratory testing, assesses the factors appropriate in introducing and managing point-of-care testing, and directs attention to realistically assessing the potential for cost effectiveness analysis. Although considered optimal, formal cost-effective analysis is limited. A commentary on the published literature of the field is presented. Observations are offered on the inadequacies of published studies that purport to describe this application. As an alternative, a more general assessment of the economic effect of point-of-care testing in the general context of the laboratory mission is recommended. Suggested guidelines are included. PMID- 7484942 TI - Guidelines for point-of-care testing. Improving patient outcomes. AB - Whole-blood biosensors and point-of-care testing created a unique paradigm in medical diagnostics in the 1980s, when liver and heart transplant centers, as well as operating rooms and other critical care areas implemented whole-blood analysis to provide rapid test results in 2 to 5 minutes. Rising expectations, patient-focused hospitals, and managed care intensify the need for immediate decisions at the point of care. The guidelines promote consensus priorities, multidisciplinary teamwork, fiscal coordination, and collaborative practice during this phase of rapid change. Four primary principles and accompanying guideline objectives are optimization (patient outcomes, medical linkages, integrated diagnostic synthesis, therapeutic turnaround time, test clusters, and critical limits), hybridization (strategic modalities, economic effectiveness, and testing --> monitoring), quality (quality improvement, clinical performance, risk reduction and academics and accreditation), and consistency (results communication and error minimization, and reference intervals and standardization). Whole-blood analysis and point-of-care testing help facilitate temporal optimization, optimize diagnostic-therapeutic processes, and improve patient outcomes in critical care. PMID- 7484943 TI - Pathologists, the death certificate, and hospital quality. PMID- 7484944 TI - Automated approaches to rapid-response testing. A comparative evaluation of point of-care and centralized laboratory testing. AB - This study evaluates the premise that point-of-care or near-patient testing provides more effective medical care because laboratory turnaround time is reduced and, therefore, presumably the time before therapy begins is reduced. An accurate evaluation of this premise must compare the operation of a point-of-care laboratory and an alternative dedicated, centralized laboratory. Assessment of both laboratory operations with respect to turnaround time, costs and benefits, regulatory factors, and finally patient care outcome are all essential in reaching a conclusion. In this study, laboratory operations in a centralized urgent care laboratory were compared with those of a point-of-care satellite laboratory. Turn-around time analysis for whole blood (heparinized specimens requiring no centrifugation) and serum analytes were measured during a representative time period. Analysis of turnaround time considered both the preanalytic time, defined as collection to receipt of the specimen in the laboratory, and the analytic time component that accounts for receipt, processing, analysis, and reporting of the final data into the laboratory information system. The preanalytic component of the point-of-care satellite laboratory was also compared with the urgent care laboratory equipped with a rapid-transport specimen system. Turnaround time in the point-of-care satellite laboratory and urgent care laboratory equipped with a rapid-transport specimen system. Turnaround time in the point-of-care satellite laboratory and urgent care laboratory equipped with a rapid-transport system were comparable. In this analysis, staff requirements, ability to comply with regulatory requirement, and the increasing demand for a larger selection of tests needed urgently were considered. In a tertiary care medical center, such as the authors', a dedicated, centralized laboratory equipped with a rapid-transport specimen system provides better comprehensive laboratory service than does a point-of-care facility with limited capability. PMID- 7484945 TI - The current and future status of critical care testing and patient monitoring. AB - Ex vivo and in vivo biosensor-based systems for patient monitoring now augment in vitro point-of-care testing, a paradigm currently in the explosive phase. As new technologies arise, so do opportunities for laboratory professionals. First, the laboratory consultant or clinical pathologist can contribute substantially to our understanding of temporal optimization and the role of diagnostic testing in optimizing diagnostic-therapeutic processes. Clarification of these facilitates wise selection of alternative testing modalities, test clusters, and instrument formats. Second, the laboratory professional is a natural member of a performance team that can help optimize outcomes and assure the quality of point-of-care testing. Explicit site-specific performance criteria for accuracy, precision, response time, and test clusters are essential and can only be established and accomplished jointly with clinicians. Third, clinical integration requires practice guidelines and care paths. These can help determine how, when, and where point-of-care testing should be implemented. Finally, global outcomes optimization calls for the input of those professionals who are most familiar with the economics of diagnostic testing, the challenges of point-of-care testing, and the best clinical benefits of in vitro, ex vivo, and in vivo biosensor-based systems in critical care and other settings. PMID- 7484946 TI - Robotics and automated workstations for rapid response testing. AB - Rapid-response testing can help the critical care physician provide more medically relevant decisions when treating critically ill patients. Many technologies have appeared on the market to help deliver rapid analytical tests, including transportation systems, hand-held analyzers, or clinical instruments that have simple user interfaces. Each of these methods can be used to provide the necessary medical information but often at the expense of turnaround time, quality of service, or cost. A robotically automated laboratory system was created that provides rapid turnaround time and low cost, and each result is monitored and reviewed by a laboratory professional. To provide the best quality laboratory services at the lowest cost, we created a remotely controlled robotic clinical laboratory that provides whole-blood analysis of blood gases (pCO2,pO2), pH, electrolytes (sodium, potassium, and chloride), glucose, and hemoglobin near the patient beside yet maintains the distinct advantage of central laboratory control. The automated remote laboratory provides extremely rapid turnaround time, eliminates the costly steps involved with specimen processing, reduces the risk from contaminated specimens, reduces staff training, ensures that every result is reviewed by a professional, and improves patient care. PMID- 7484947 TI - The satellite laboratory and point-of-care testing. Integration of information. AB - Laboratory testing is returning to the point of care, where it started in the 19th century. This decentralization of testing is occurring because of the many advantages it brings to the practice of critical care medicine, such as faster turn-around time and better patient management in inpatient and outpatient settings. However, decentralization of testing is decentralizing laboratory information. In addition, bedside analytical systems are causing a quality assurance dilemma in which data can be seen and acted on before the appropriate quality checks are completed. Current information technology allows these data to be integrated and thus patient care processes to be enhanced through tracking of clinical outcomes and reducing costs. Because the goal of point-of-care testing is to provide data in a timely manner, the future success of any point-of care/satellite testing system will depend on the ease with which it can be integrated into the medical information system. PMID- 7484948 TI - Regulatory requirements (CLIA '88, JCAHO, CAP) for decentralized testing. AB - All decentralized testing is regulated according to the Clinical Laboratory Improvement Amendments of 1988. Two organizations, the Joint Commission on Accreditation of Health Care Organizations and the College of American Pathologists, have received deemed status for their voluntary standards from the Health Care Financing Administration. Deemed status means that the organizations' voluntary standards meet or exceed the federal requirements. The decentralized testing sites can exercise several options in determining which organization, and hence which set of standards, will be used to regulate their testing processes. In this article, the authors outline the various regulatory requirements, provide insight into the relationship of each, and offer a framework for decentralized testing sites to follow to meet the requirements. PMID- 7484949 TI - Managed health care. Managing the risks of laboratory testing in an integrated delivery system. AB - Decentralized testing must be viewed as a component of the decentralized patient care that is prevalent today. It cannot be considered a separate component of the total laboratory process, but must be examined in the context of test ordering and test performance through test result disposition and use. The evolving configuration of the health-care delivery system necessitates a redefinition of risk to include four categories of risk: business, financial, insurable, and clinical quality. Reengineering the clinical laboratory process in today's health care environment needs to incorporate all these risk issues, systems thinking, and available technology along with the changing practice of medicine. PMID- 7484950 TI - Airborne and rescue point-of-care testing. AB - The prehospital environment has not been studied for point-of-care testing. Therefore, the authors' helicopter program evaluated the performance of the i STAT Portable Clinical Analyzer, a rapid point-of-care, hand-held instrument. The primary aim of the study was to determine if the i-STAT Portable Clinical Analyzer could be used in the field to assess patient status in flight, and to allow the flight crew to intervene immediately, thus delivering a more stable patient to the emergency room. Imprecision and initial split-sample comparative studies with the Portable Clinical Analyzer were completed in the hospital satellite laboratory and clinical chemistry laboratory. Comparison studies were performed on patient samples drawn and analyzed in the helicopter and subsequently analyzed in the satellite and clinical chemistry laboratories. The only significant differences observed were with glucose. The glucose discrepancies were probably due to the time delay between collection of the specimen in the helicopter and subsequent analysis in the laboratory. Following this initial validation, the i-STAT Portable Clinical Analyzer was used on 81 patients transported by the flight crew. The tests performed in the helicopter include sodium, potassium, glucose, and hematocrit/hemoglobin concentrations. Fifteen (18.5%) of the patients were treated with transfusions, glucose, or insulin based on the Portable Clinical Analyzer results. Other identified needs include blood gas analysis (in process) and use of point-of-care testing in the fixed-wing environment. PMID- 7484951 TI - Advances and strategies for glucose monitoring. AB - Blood glucose monitoring systems are used effectively in the therapy of persons with diabetes mellitus. Through the simple application of a drop of blood, a person can immediately obtain a blood glucose concentration. Many systems are being marketed and sold worldwide. Therefore it is essential that the patient and health-care professional understand the similarities, differences, and use characteristics of these systems. Systems to self-monitor blood glucose levels are used in three distinct areas. Area 1 is the patient home use setting, area 2 is the physician's office or diabetes clinic setting, and area 3 is the hospital or medical center setting. Each area has associated with it certain challenges that are distinct from the others. Strategies to meet these challenges are explored along with ways to assess confidently the performance of any self monitoring blood glucose system in each of these areas. PMID- 7484952 TI - Thrombosis and hemostasis testing at the point of care. AB - Point-of-care testing provides current, accurate information relating to thrombosis and hemostasis in patients. Several forces are driving point-of-care testing, particularly economic factors and improved technology. Point-of-care testing has the potential to improve patient management and to decrease integrated costs, although this remains to be shown. For such testing to be successful, the technology must be complemented by hospital-wide point-of-care testing programs. The role of the laboratorian and pathologist will become important for coordinating programs, maintaining quality assurance, and promoting quality improvement. PMID- 7484953 TI - Critical care profiling for informed treatment of severely ill patients. AB - An important advance in critical care medicine is the availability of key biochemical test results at the patient's bedside within the brief period when they can be used to guide resuscitation and cardiovascular stabilization effectively. Frequently the menu of key tests, called a "critical care profile (CCP)," must be available to the attending clinician within 5 minutes to guide therapy in real time. Recent developments that make it possible to deliver a CCP with these 5 minutes include the ability to measure all of the analytes of the critical care profile on a whole-blood sample, and point-of-care testing. Today the results of a critical care profile can be available to the attending clinician within 5 minutes of his or her request by sampling approximately 200 microL of whole blood with a critical care instrument at or near a patient's bedside. Tests frequently recommended for a critical care profile include glucose, pO2, pCO2, pH, lactate, ionized calcium, potassium, sodium, ionized magnesium, and either hemoglobin concentration or hematocrit. Hospital areas that need critical care profile support include the emergency department, the operating room, and the intensive care unit. PMID- 7484954 TI - Physiology and regulation. Ionized calcium, magnesium and lactate measurements in critical care settings. AB - The biochemical and physiologic bases for clinical use of ionized calcium, magnesium, or lactate measurements are described, with emphasis on the critical care setting. Ionized calcium or magnesium are usually measured to prevent hypocalcemia or hypomagnesemia, conditions that most affect cardiovascular function. Blood lactate measurements have been used more frequently in the last 5 years, despite decreases in overall laboratory test volume. Lactate measurements appear to have clinical value, particularly for children having major cardiac surgery or extracorporeal membrane oxygenation. The changes in test volumes in the past few years due to the changing economic climate in medical practice are also described. PMID- 7484955 TI - Point-of-care testing in the intensive care unit. The intensive care physician's perspective. AB - Technological advancements have, for the first time, made the entire laboratory testing process feasible at the bedside. Physicians working in the intensive care unit have always had immediate access to patients' medical history, physical examination, and physiologic monitoring data, but had to wait for laboratory results. Using point-of-care testing, laboratory parameters targeted to critical illnesses can now be integrated into initial diagnostic assessments, on patient rounds, and during therapeutic maneuvers. The concept of point-of-care testing in the intensive care unit is relatively new, but as technology progresses, physicians will undoubtedly become aware and use it in the intensive care unit. This article focuses on the intensive care physician's perspective on laboratory testing, the evolution of the intensive care unit laboratory, advantages of point of-care testing in that setting, new developments in arterial blood gas analyzers and monitors, and cost-effectiveness and incorporation of point-of-care testing. PMID- 7484956 TI - Albert H. Ketcham Award recipients for 1996 announced by the ABO. PMID- 7484958 TI - Standards, indices, and managed care. PMID- 7484957 TI - Comment on obstructive sleep apnea. PMID- 7484959 TI - Cosi fan tutti! PMID- 7484960 TI - Traveling minstrels. PMID- 7484961 TI - Historic publication on the first use of magnets in orthodontics. PMID- 7484962 TI - Comment on the Brodie bite. PMID- 7484963 TI - Comment on biomechanics of the three-piece base arch appliance. PMID- 7484964 TI - Interlayer formation and its effect on debonding polycrystalline alumina orthodontic brackets. AB - During the debonding of ceramic orthodontic brackets, there is a risk of causing fractures, cracks, or flaking of enamel or of the bracket itself. Preliminary work on the resin-enamel interface under bonded brackets with the indirect (modified Thomas) or the thermal-cured indirect bonding techniques revealed an interlayer of unfilled resin formed between the filled resin and the enamel surface. The direct bonding technique, on the other hand, showed no such layer. This study was designed to determine the effect of the interlayer on conventional debonding techniques for polycrystalline ceramic orthodontic brackets. Variables examined were bracket failure or fracture (BF), amount of remnant adhesive (ARI), and enamel damage. Brackets were bonded to 90 fresh bovine teeth. These were divided into three groups of 30 each, based on three methods of bonding, i.e., direct, indirect (modified Thomas), and an indirect technique that used a thermal cured resin. Each bonding group was further divided into three groups of 10 each, based on the type of debonding technique used, i.e., lift off, delamination, and twisting. Brackets bonded by the indirect (modified Thomas, BF mean = 0.27, ARI mean = 0.93) and the indirect technique that used a thermal-cured resin (BF mean = 0.03, ARI mean = 0.43) resulted in an overall significantly lower failure (p < 0.01) and ARI score on debonding (p < 0.0001) compared with those bonded by the direct technique (BF mean = 1.03, ARI mean = 1.97). Specimens evaluated under the stereomicroscope revealed that the brackets bonded with the indirect techniques debonded at the filled-unfilled resin interface or within the interlayer of unfilled resin.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7484965 TI - Multiflex versus superelastic: a randomized clinical trial of the tooth alignment ability of initial arch wires. AB - Two arch wires commonly used for initial tooth alignment were compared with regard to their clinical effectiveness. The two arch wires tested were 0.0155 inch diameter multiple-stranded stainless steel wire (Dentaflex, Dentaurium, Optident, Yorkshire, England) and 0.014-inch diameter nickel-titanium alloy wire (NiTi, ORMCO Co., Monrovia, Calif.). Consecutive patients attending an orthodontic clinic for routine placement of a fixed appliance were randomly assigned one of these two initial arch wires. Good quality alginate impressions of the appropriate dental arch were taken before arch wire placement and also at the subsequent appointment, which was, on average, 6 weeks later. Seventy-four arches were used in this study. The degree of tooth alignment achieved for each wire type was compared with a Reflex Microscope (Reflex Measurement Ltd., Butleigh, England) to make detailed measurements on the resultant casts. The degree of initial alignment achieved with the two wires was similar over this 6 week period. However, some differences were found for the lower labial segment where the interbracket span is usually reduced and where the superelastic nickel titanium wire was found to give improved alignment. No threshold of crowding was found where one arch wire performed better than the other. PMID- 7484966 TI - Craniofacial structure and soft tissue profile in patients with severe hypodontia. AB - This study compares craniofacial structure and soft tissue profile in persons with mild hypodontia (group I: 2 to 5 congenitally missing teeth, n = 43), moderate hypodontia (group II: 6 to 9 congenitally missing teeth, n = 15) and severe hypodontia (group III: 10 or more congenitally missing teeth, n = 29) with the structure of persons without hypodontia and with normal occlusion (n = 50). The mean age was about 12 years. In group I, the lower second premolars were the most frequently missing teeth, followed by the upper second premolars and the upper lateral incisors. The relative prevalence of missing second premolars decreased with increasing severity of hypodontia. No consistent pattern could be observed when more than five teeth were missing, indicating a different genetic mechanism than for mild hypodontia. A significant retroclination of the incisors and an increased interincisal angle were observed with increasing severity of hypodontia. This was accompanied by a reduction of lip protrusion, being most evident for the upper lip. Increasing numbers of missing teeth resulted also in a decrease in the mandibular plane angle and a reduction in the anterior lower facial height. Few differences in the skeletal parameters were observed. It was concluded that the typical dentofacial structure in persons with advanced hypodontia may be due to dental and functional compensation rather than to a different growth pattern. PMID- 7484967 TI - Long-term stability of rapid palatal expander treatment and edgewise mechanotherapy. AB - Previous studies on long-term stability of orthodontic treatment primarily have focused on the stability of the lower arch treated with edgewise appliances. The aim of this study was to evaluate the long-term stability of the upper and the lower dental arches of patients treated with a rapid palatal expander. The sample comprised of 165 dental casts randomly selected from patients who had been out of retention for 8 to 10 years at a mean age of 30 years. Measurements were made directly on dental casts obtained at the three time intervals: before treatment, after treatment, and after retention. Differences over time between the upper and the lower dental arches and between intervals were analyzed by a two-way multivariate analysis of variance (MANOVA) and post hoc Bonferroni t tests. Differences between after treatment and after retention were statistically significant (P < 0.006) for all except lower intermolar width. However, only for lower and upper arch lengths and perimeters were the differences greater than 2.0 mm. Treatment with the rapid palatal expander presented good stability for upper intercanine width, upper and lower intermolar widths and incisor irregularity. Lower intercanine, arch length, and perimeter presented poor stability. PMID- 7484968 TI - Changes in facial dimensions assessed from lateral and frontal photographs. Part II--Results and conclusions. PMID- 7484969 TI - The effect of bite-blocks with and without repelling magnets studied histomorphometrically in the rhesus monkey (Macaca mulatta). AB - The effect of bite-blocks with and without repelling magnets as proposed for the treatment of open bite was analyzed. Twelve male juvenile monkeys were divided into three groups of four. Group A was used as control, group B was given bite blocks containing samarium cobalt disks, and group C received identical bite blocks without active magnets. The monkeys were observed for 24 weeks before death. Histomorphometric evaluation was then performed on the molar roots, their periodontal tissues, the zygomaticotemporal suture, and the pterygomaxillary suture. The root surfaces of the molars in both the bite-block group and the magnetic group were characterized by pronounced resorption that sometimes was active and occasionally undergoing repair with bony tissue. The sutures also clearly reflected the effect of both appliances used, although more markedly in the cases of bite-blocks containing active magnets. The surface density expressing the sutural area, was increased significantly, possibly as an adaptation to the altered functional demand. The cellular activity of the sutural surfaces also was increased markedly in both appliance groups, reflecting an ongoing adaptation. A steady state had not been reached. The study demonstrated a widespread effect of the force developed by bite-blocks with and without magnets. The final quantity and the reversibility of the effect is not known, however. More long-term studies should be undertaken to obtain this information. PMID- 7484970 TI - Improving orthodontic bonding to gold alloy. AB - Flat tabs of cast gold alloy (n = 156) were subjected to either of three surface treatments: (1) roughening with diamond bur, (2) aluminum oxide sandblasting, and (3) sandblasting plus tin electroplating. Mandibular incisor edgewise brackets were bonded with Concise (BIS-GMA resin) (Unitek, Monrovia, Calif.) or Superbond C&B (4-META metal bonding resin) (Sun Medical Co. Ltd., Kyoto, Japan), or with Concise after application of an intermediate resin. All-Bond 2 Primers A and B (Bisco Dental Products, Itasca, Ill.), or B alone. All specimens were stored in water at 37 degrees C for 24 hours, and 60 were then thermocycled 1,000 times from 5 degrees C to 55 degrees C and back. The tensile bond strength testing was performed in a Lloyd 1,000R machine (Fareham, Hants, England). Alignment and uniform loading during testing were secured by engaging a hook in a circular ring soldered onto the bracket slot before bonding. Similar control brackets (n = 24) were bonded with Concise to extracted human premolars and lower incisors according to a routine procedure. Bond failure sites were classified by a modified ARI system. The results showed that sandblasting produced significantly stronger bonds to gold alloy than roughening with diamond bur. Superbond C&B provided significantly stronger bonds to gold alloy than Concise. There were generally insignificant differences in bond strengths between the water stored and the thermocycled specimens. Bond failures of Concise to sandblasted plus tin plated gold alloy invariably occurred at the gold/adhesive interface, whereas those of Superbond C&B occurred within the adhesive or in the adhesive/bracket interface.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7484972 TI - Mandibular skeletal changes induced by early functional treatment of Class III malocclusion: a superimposition study. AB - A superimposition study on mandibular stable structures was performed to evaluate changes in mandibular rotation and in the direction of condylar growth induced by early treatment of Class III malocclusion by a functional appliance (removable mandibular retractor). A sample of 18 children with treated Class III malocclusions, mean age at the first observation (immediately before the beginning of treatment) 5.47 +/- 1.14 years, mean age at the second observation 8 +/- 1.29 years, mean observation period 2.5 +/- 0.9 years, was compared with a control group of 18 children with untreated Class III malocclusions, mean age at the first observation 5.72 +/- 1.11 years, mean age at the second observation 7.86 +/- 1.44 years, mean observation period 2.27 +/- 0.92 years. No statistically significant difference between the two groups was recorded for positional (total) rotation of the mandible. A significant upward-forward direction of condylar growth was assessed in the treated group (p < 0.001). This therapeutically induced change in growth direction of the mandibular condyle was considered a skeletal sign of anterior morphogenetic rotation of the mandible, i.e., a mechanism compensating for excessive mandibular growth. PMID- 7484971 TI - Orthodontic forces increase tumor necrosis factor alpha in the human gingival sulcus. AB - The production of cytokines has been associated with the biology of tooth movement in animal populations. The purpose of this study was to measure tumor necrosis factor-alpha (TNF) directly in the human gingival sulcus before and after the application of an orthodontic force. To recover TNF from the sulcus, paramagnetic beads, coated with monoclonal antibodies for TNF, were introduced into the gingival sulcus of 50 teeth undergoing orthodontic tooth movement (by two force systems) in 20 patients. Retrieval was performed by a permanent magnetic device designed to fit the periodontal sulcus. The samples were taken before force application (controls), and at a fixed time after force application. The amount of immunoabsorbed TNF was quantified with an immunochemical assay. There was a greater than twofold increase in TNF recoverable from the gingival sulcus after application of orthodontic forces (mean of 12.9 ng vs 30.5 ng). A Student's t test for paired samples demonstrated statistical significance at p < 0.01. We conclude that the quantity of paradental TNF, found in human gingival sulcus, is elevated during tooth movement. The source may be from the adjacent gingiva, but more likely the compressed periodontal ligament and the resorbing bone adjacent to the root surface. PMID- 7484973 TI - Extraction of a mandibular incisor in a Class I malocclusion. AB - A single mandibular central incisor was removed to treat a Class I malocclusion with crowding and a midline discrepancy. This case report was submitted under category number 10 (optional) and it was believed that the results illustrated the Board objectives of (1) facial harmony, soft tissue balance, and proper proportion, (2) maximum health of the teeth, the supporting tissues and adjacent structures, (3) maximum esthetics, (4) optimal functional relationships, (5) treatment in harmony with the patient's facial growth, and (6) stability. Beginning, completion, and 2-year retention records are included. The text follows the guidelines established by the Board for content and length included in "Specific Instructions for Candidates." [This case report was presented to the American Board of Orthodontics in partial fulfillment of the requirements for the certification process conducted by the Board.] PMID- 7484974 TI - Treatment of a patient with a mutilated Class II, Division 1 malocclusion and a dolichofacial skeletal pattern. AB - The purpose of this case report is to present a description of the diagnosis and treatment to ABO standards of a patient with a mutilated Angle's Class II, Division 1 malocclusion, complicated by anterior dental crowding, and a dolichofacial skeletal pattern. Treatment involved extraction of the maxillary first premolars, lingual arches to enhance anchorage, and J-hook headgear to retract the maxillary anterior segments. PMID- 7484975 TI - Case reports: molar distalization with static repelling magnets. Part II. AB - The following two nonextraction therapy case reports demonstrate four important points: (1) Static repelling magnets, with certain characteristic parameters, distalize molars rapidly without adverse effects that are clinically discernable. Beneficial properties such as considerably reduced patient compliance requirements for force application, reduced mobility and discomfort, and mostly bodily movement are demonstrated clinically. (2) Anchorage can be controlled by using conventional techniques of anchorage augmentation or force reduction. (3) Treatment can be satisfactorily completed and documented in accord with contemporary criteria, with conventional therapeutic methods, once the initial, most difficult, magnetic molar distalization is completed. (4) The mechanism of action that accounts for the bioeffects remains unclear. However, we hypothesize that observed reduction of mobility and discomfort during rapid movement, combined with basic research and other clinical data, are attributable to the simultaneous, synergistic property of the magnetic force field, which disrupts the local equilibrium and also the static magnetic bioeffect. Among others, one distinguishing and pertinent bioeffect may be an increased rate of osteogenesis and bone remodeling, which may be very dependent on correct dosage through a possible biologic window and the field geometry. PMID- 7484976 TI - One-phase versus two-phase treatment. PMID- 7484978 TI - Computerized archiving of records. PMID- 7484977 TI - Efficient orthodontic treatment timing. PMID- 7484979 TI - Duty to follow up on recommended care. PMID- 7484980 TI - Craving in hospitalized cocaine abusers as a predictor of outcome. AB - Despite its common use, the term "craving" is controversial. Moreover, its measurement can be problematic, since craving is purely subjective and is highly influenced by setting and drug availability. Thus, one might expect patients in protective settings such as hospitals to experience little craving, despite potentially being at high risk to relapse upon discharge. In this study of 73 patients hospitalized for cocaine dependence, we developed a series of five questions to measure different aspects of cocaine craving: 1) current intensity, 2) intensity during the previous 24 hours, 3) frequency, 4) responsiveness to drug-related conditioned stimuli, and 5) imagined likelihood of use if in a setting with access to drugs. We hypothesized that some hospitalized patients with low craving levels might believe their risk of relapse to be high in another setting. We then examined the relationship between inpatient craving levels and 3 month treatment outcome. Results revealed a high level of internal consistency, with Cronbach's alpha scores of .82 to .94 on daily measures; factor analysis revealed unidimensionality; and each of the five items showed significant decreases in craving over time. The question regarding likelihood of use in another setting consistently scored higher than other questions on current craving (p < .001). Predictive validity was assessed by examining 3-month outcome. Among the 37 patients who participated in a follow-up study, there were no statistically significant relationships between 3-month cocaine use outcome and any item on the craving scale. Neither inpatient craving levels nor predictions regarding future use correlated with actual 3-month outcome. PMID- 7484981 TI - Effects of psychiatric care for dual diagnosis patients treated in a drug dependence clinic. AB - This study examines outcome of treatment for psychoactive substance dependence in a clinic which made psychiatric care readily available. Veterans entering outpatient treatment for substance dependence (n = 222) received psychiatric evaluation for additional Axis I disorders using DSM-III-R criteria. Patients provided urine toxicology specimens at least weekly. Outcome (urinalysis results and treatment retention) was compared for patients with dual diagnosis (n = 103, 46.4%) and with substance only diagnosis (n = 119, 53.6%). Psychotropic medications were prescribed for 80.4% of the dual diagnosis subjects. In the first 6 months of treatment, dual diagnosis subjects compared to substance only diagnosis subjects gave a significantly greater percentage of urines positive for cocaine and opioids. In the second 6 months, dual diagnosis subjects who remained (n = 72, 70.0%) significantly reduced from the first 6 months their percentage of cocaine and opioid positives and did not differ in percent positives from substance only diagnosis subjects who continued past 6 months (n = 70, 58.8%). Treatment retention of dual diagnosis subjects (median months = 14.3) exceeded that of substance only diagnosis subjects (8.9; Lee-Desu Statistic = 9.02, p < .003). Dual diagnosis patients may initially perform more poorly than substance only diagnosis patients in substance dependence treatment. However, in the presence of psychiatric care, they eventually exhibit comparable success. PMID- 7484982 TI - Dimensions of self-concept: a comparison of heroin and cocaine addicts. AB - Two groups of drug users on an inner-city inpatient drug detoxification unit were studied: 42 heroin addicts and 47 cocaine addicts. The two groups were compared on personality disorder diagnoses, personality traits, and demographic variables. Cocaine and heroin addicts scored similarly on: 1) number and kind of personality disorder diagnoses, with the exception of antisocial personality; 2) all personality traits measured; 3) positive and negative temperament; 4) description of self-concepts; and 5) positivity and negativity of self-concepts. It was also found that heroin addicts showed significantly higher levels of social deviance than the cocaine group, with significantly more antisocial personality disorder diagnoses, higher levels of social deviance, and lower scores on a Propriety Scale. Character pathology was more heterogeneous among cocaine users. Heroin addicts had used significantly longer and showed less educational and occupational achievement than the cocaine group. Our results also suggest that personality psychopathology in drug addicts is associated with lower self-esteem, more negative self-valuation, and longer duration of use. PMID- 7484984 TI - Suicidality in a sample of methadone maintenance clients. AB - Previous work has shown that suicide is a significant cause of death among substance abusers, including methadone-maintained clients, and that the prediction of suicidal ideation and behavior is difficult. Preliminary review of data collected at admission on a population of 438 methadone-maintained clients found 55 expressing some level of suicidal behaviors during the course of treatment. These clients were compared with a randomly selected comparison group of 55 nonsuicide clients matched for gender and race/ethnicity on measures of psychological dysfunction, drug use, family dysfunction, and help-seeking behaviors. Results showed that at time of admission suicidal clients reported: 1) more psychological dysfunction as evidenced by higher levels of depression, social dysfunction, hostility, risk-taking, and previous thoughts of suicide; 2) less family support at the present time and during childhood; and 3) more help seeking behavior as evidenced by self-referral, number of previous treatment episodes, attendance at self-help meetings, and higher scores on motivational measures of desire for help. Differences in preadmission drug-using behaviors were not found between the two groups. PMID- 7484983 TI - Effects of combined fluoxetine and counseling in the outpatient treatment of cocaine abusers. AB - Three methods of analysis were used to determine the effects of the combination of counseling with fluoxetine (20, 40, or 60 mg) and "active" placebo (diphenhydramine, 12.5 mg) randomly assigned. Forty-five cocaine-only dependent subjects were treated as outpatients with "interpersonal" counseling, medication, and drug use monitoring three times per week for up to 12 weeks. Treatment effects were analyzed: first, by comparing the three original assignments and placebo; second, by comparing the placebo group to fluoxetine subjects with detectable fluoxetine/norfluoxetine blood levels and those with no detectable medication blood level; third, by examining relapse prevention versus use cessation through stratifying the subjects into four groups according to fluoxetine or placebo assignment and initial urine cocaine positivity or negativity. All three analyses showed improvement on some measures over time regardless of group assignment. The 60-mg fluoxetine group showed least effectiveness, the group with detectable blood levels had less cravings, and the fluoxetine subjects who were abstinent at the start of treatment were somewhat less likely to avoid relapse than those on placebo. PMID- 7484985 TI - Homovanillic acid and dopamine-beta-hydroxylase in male youth: relationships with paternal substance abuse and antisocial behavior. AB - Recent research suggests that dopaminergic/noradrenergic system dysfunction may be associated with substance abuse and/or antisocial behavior. In order to determine whether male youth of fathers with these disorders would manifest differences in these systems when compared with youth of nonsubstance-abusing or nonantisocial fathers, levels of homovanillic acid (HVA), the metabolite of dopamine (DA) and dopamine-beta-hydroxylase (DBH), the enzyme facilitating the conversion of dopamine to norepineprhine, were studied in offspring blood samples. The subjects were 65 male youth aged 6-15 years admitted to a residential center because of behavioral disorders. Parental substance abuse and antisocial behavior were assessed through interviews, rating scales, and/or chart review. HVA and DBH were determined from blood samples obtained after admission. The findings indicated that youth of substance-abusing fathers had significantly greater levels of HVA than yough of nonsubstance-abusing fathers. Younger (< 12.0 years) boys of antisocial fathers had significantly lower DBH activity than comparably aged youth of nonantisocial fathers. The results suggest that common generational links in substance abuse and antisocial behavior in males may be associated with detectable biological parameters in susceptible youth. PMID- 7484986 TI - Effects of halfway house placement on retention of patients in substance abuse aftercare. AB - One hundred twenty-four male veterans were enrolled in the Birmingham VAMC Outpatient Substance Abuse Clinic (OSAC) aftercare program following inpatient treatment for alcohol, cocaine, or mixed alcohol-cocaine abuse/dependence. Forty two of the patients were concurrently admitted to a nearby halfway house (HH) while the remaining 82 patients made community-based (CB) living arrangements. Chi-square analysis showed the two groups were matched, p > .05, in regard to age, race, marital status, presenting problem, and referral source. The CB group experienced significantly, p < .05, higher early dropout (40 vs 0%) from aftercare. Of the others engaging in treatment, the HH patients stayed in OSAC aftercare 60 days longer and had significantly, p < .01, more clinic visits. A higher proportion, p < .01, of HH patients completed each of four treatment milestones: education group, psychological testing, psychological interview, and treatment planning/update. On average, the HH patients remained in OSAC an additional 90 days after their halfway house discharge. Although not significantly different, p > .05, the frequency of HH patients receiving administrative discharge status of Treatment Complete was twice (28.2%) that of CB patients (15.1%). It is concluded that concurrent halfway house placement can aid in aftercare retention and completion. PMID- 7484987 TI - The effects of travel barriers and age on the utilization of alcoholism treatment aftercare. AB - OBJECTIVE: The objective of this research was to ascertain whether geographical accessibility (in conjunction with other patient characteristics) reduced the probability of participating in alcoholism aftercare treatment. METHODS: A sample of 4,621 United States male veterans discharged with an outpatient appointment from one of 33 Department of Veterans Affairs inpatient Alcohol Dependency Treatment Programs was identified. The outpatient records of each patient were obtained to determine whether aftercare services were utilized following discharge. Binary choice analysis was used to model the decision to enter aftercare treatment as a function of travel distance, age, marital status, ethnicity, severity of illness, and urbanization. RESULTS: Travel barriers significantly reduced aftercare participation, especially for elderly and rural veterans. Both younger and older veterans were less likely to keep their aftercare appointment than middle-aged veterans. Married patients were more likely to utilize outpatient services than unmarried patients. Ethnic status, severity of illness, and urban size all negatively affected the likelihood of appointment attendance. CONCLUSIONS: The results obtained from this analysis can be effectively used to identify which patients are not likely to enter aftercare alcoholism treatment. The discharge plans of patients at risk for appointment noncompliance should be given special attention since aftercare has been shown to improve treatment outcome. Moreover, because alcoholism treatment reduces the utilization of other medical services, promoting continuity of care should help lower the overall costs of providing health care to alcoholic patients. PMID- 7484988 TI - Alcohol availability and alcohol-related crashes: does distance make a difference? AB - Variations in alcohol control laws can cause problems if individuals travel to less restrictive jurisdictions to purchase alcohol to circumvent more restrictive regulations. The establishment of a national minimum drinking age of 21 is an example of an attempt to eliminate the phenomenon known as "border drinking." However, it exists today in many southern states that have dry counties. The present study analyzes Kentucky's 77 dry counties to determine if rates of six types of motor vehicle crashes are affected by distance to legal alcohol. The analysis indicates that the distance variable is significantly and negatively related to rate of alcohol-related injury crashes but does not appear to be a substantial determinant of accident rates in dry counties. PMID- 7484989 TI - Transjugular intrahepatic portosystemic shunts: is TIPSS definitive therapy? PMID- 7484990 TI - Infectious esophagitis in AIDS: what have we learned in the last decade? PMID- 7484991 TI - The tortoise and the hare: the race toward improved treatment for gallstones. PMID- 7484992 TI - Results of a survey to evaluate whether vaginal delivery and episiotomy lead to perineal involvement in women with Crohn's disease. AB - OBJECTIVE: Crohn's disease frequently affects the perineum, but it is unknown if such involvement poses a heightened risk to a woman with Crohn's disease undergoing vaginal delivery and possibly episiotomy. This study attempts to elucidate whether vaginal delivery with or without episiotomy: 1) predisposes to perineal involvement in women with Crohn's disease; 2) reactivates quiescent perineal Crohn's disease; or 3) worsens preexisting perineal Crohn's disease. METHODS: Data were culled from questionnaires returned by 117 respondents contacted through newsletters of the Crohn's and Colitis Foundation of America. These surveys were then reviewed and tabulated to determine if any temporal and causal relationship existed between perineal Crohn's disease and vaginal delivery. RESULTS: Four groups were identified: group 1 (n = 56), with 128 pregnancies, had no history of perineal disease or subsequent perineal complication. Group 2 (n = 24) had 51 pregnancies. This group had no preexisting perineal disease, but 67.6% said they developed perineal Crohn's disease postpartum, 60% of which occurred within 2 months of vaginal delivery. Groups 3 and 4 were too small to evaluate (n = 4 and 5, respectively) and represented patients with preexisting perineal disease who had varying results after delivery. Overall, the rate of developing perineal involvement after vaginal delivery, usually with episiotomy, in patients with Crohn's disease and no preexisting perineal involvement (combining groups 1 and 2) was 17.9%. CONCLUSIONS: A high rate of perineal involvement seems to follow vaginal delivery with episiotomy in patients with Crohn's disease. Although this study has substantial methodological limitations, it does raise questions as to the proper obstetrical management of patients with Crohn's disease and highlights an intriguing relationship that deserves further and more rigorous study. PMID- 7484993 TI - Nonsurgical treatment of variceal bleeding: new modalities. PMID- 7484994 TI - A comparative study of emergency transjugular intrahepatic portosystemic stent shunt and esophageal transection in the management of uncontrolled variceal hemorrhage. AB - OBJECTIVES: Transjugular intrahepatic portosystemic stent-shunts (TIPSS) have been shown to reduce portal hypertension consistently and have recently been reported to arrest active variceal hemorrhage. This retrospective and nonrandomized study compares the results of TIPSS with esophageal transection (ET) and devascularization in patients with uncontrolled variceal hemorrhage admitted to a single center with an interest in variceal bleeding. PATIENTS AND METHODS: Two hundred and sixty cirrhotic patients have been referred with variceal bleeding over the past 7 yr. In 41 patients (15.8%), hemorrhage was uncontrolled despite two treatments with sclerotherapy. Thirty-eight patients were eligible for analysis. Nineteen were considered for ET and 19 for TIPSS. Patients in the two groups were well matched for age, sex, etiology of liver disease, and its severity and complications. They have been followed for 13 patient years (TIPSS-7, longest 20 months; ET-6, longest 23 months). Data for survival and rebleeding were analyzed by the Kaplan-Meier method on an intention to-treat basis. RESULTS: Seven of the 19 were considered unfit for surgery, and 12 underwent esophageal transection and devascularization. TIPSS was undertaken successfully in 17 patients, the Palmaz stent being used in 4 and the Wallstent in 13. Successful TIPSS reduced the mean portal pressure gradient from 22.2 (SE 1.2) to 9.7 (SE 0.7) mm Hg (p < 0.001). Mortality within 30 days of the initial bleed was 42% in the TIPSS group compared with 79% in the ET group (p < 0.05). Rebleeding occurred in 15.6% patients with TIPSS, compared with 26.2% in the ET group. Encephalopathy in the two groups of patients was not significantly different (TIPSS 25% and ET 22%). TIPSS was followed by active infection in 20% compared with 36% after ET. CONCLUSIONS: This study shows that the overall mortality in this group of patients is high whatever the type of treatment used. TIPSS can be performed successfully on these patients who are often not suitable for surgery. Mortality rates were significantly lower in patients treated by TIPSS. Compared with ET, TIPSS should be regarded as the preferred mode of treatment for uncontrolled variceal hemorrhage in patients with cirrhosis. PMID- 7484995 TI - Prospective evaluation of oropharyngeal findings in human immunodeficiency virus infected patients with esophageal ulceration. AB - OBJECTIVE: Although the presence of oropharyngeal (OP) candidiasis plays an important role in the evaluation of human immunodeficiency virus (HIV)-infected patients with esophageal symptoms, there is little information on the utility of OP findings in patients with esophageal ulceration. METHODS: Over a 54-month period, all HIV-infected patients with esophageal ulceration had careful inspection of the oropharynx at the time of presentation with esophageal complaints and at endoscopy. HIV-infected patients without esophageal ulceration undergoing endoscopy during the last 30 months had OP findings similarly documented. OP ulceration ulceration was determined based on clinical, endoscopic, and histopathological findings. RESULTS: Of the 124 patients identified with esophageal ulcer, 14 (11%) had coexistent OP ulceration: herpes simplex virus, four; idiopathic esophageal ulcer, four; cytomegalovirus, three; herpes simplex virus and cytomegalovirus, two; idiopathic and cytomegalovirus, one. Four patients had OP ulcer without esophageal ulcer; only one of these patients had esophageal symptoms. All OP lesions healed with therapy for the esophageal ulcer. Twenty-eight patients with esophageal ulcer had OP candidiasis (23%); 21 of these patients (75%) also had Candida esophagitis. The sensitivity and specificity of OP ulcer for esophageal ulcer were 11% and 97%, respectively. The positive and negative predictive values of OP candidiasis for esophageal candidiasis were 90% and 82%, respectively. CONCLUSIONS: OP ulceration is uncommon in patients with esophageal ulceration, with the exception of herpes simplex virus esophagitis. OP candidiasis is common in patients with underlying esophageal ulcer, potentially resulting in diagnostic confusion. OP candidiasis appears to be a moderately useful diagnostic marker for Candida esophagitis. PMID- 7484996 TI - Combined bile acid therapy is more effective on biliary lipids and dissolution rates than monotherapy after gallstone lithotripsy. AB - BACKGROUND: Accurate sampling of gallbladder bile for biliary analysis is essential for determining any potential difference between combined bile acid therapy and monotherapy in gallstone patients. METHODS: In 104 gallstone patients undergoing extracorporeal shock wave lithotripsy with following bile acid therapy [either chenodeoxycholic acid (500 mg/day) and ursodeoxycholic acid (500 mg/day), group I (n = 53), or ursodeoxycholic acid alone (1000 mg/day), group II (n = 51)], bile samples, obtained by direct fine needle puncture of the gallbladder, were investigated for biliary lipids, total biliary protein concentration, and nucleation time before and after 12 months of bile acid therapy. RESULTS: Initially, a negative correlation was found between nucleation time and number of gallstones and between total biliary protein concentration and nucleation time (r = -0.52 and r = -0.49 in group I vs r = -0.56 and r = -0.51 in group II, p < 0.01 in each group). The correlation between total biliary protein concentration and nucleation time was also found after 12 months of bile acid treatment (r = -0.54 in group I vs r = -0.47 in group II, p < 0.01 in each group). In group I, the decrease in cholesterol saturation index, biliary cholesterol, cholic acid, deoxycholic acid, and total protein concentration was more pronounced than in group II (p < 0.01). The same effect was found concerning the prolongation of nucleation time (p < 0.01). Furthermore, dissolution rates were higher in group I compared with group II (80.4 vs 69.0%, p < 0.01). CONCLUSION: In gallstone patients, combined therapy with urso- and chenodeoxycholic acid is superior to either ursodeoxycholic acid alone or biliary parameters in bile samples obtained by direct fine needle puncture of the gallbladder. PMID- 7484997 TI - Remaining small bowel endoscopic lesions at surgery have no influence on early anastomotic recurrences in Crohn's disease. AB - OBJECTIVES: We recently demonstrated that 65% of patients operated on for Crohn's disease (CD) had lesions of the small bowel at perioperative endoscopy (POE). These lesions were unrecognized before surgery in more than half of the patients. The aim of this study was to assess the prognostic value of endoscopic small bowel lesions let in place at time of surgery on further anastomotic endoscopic relapse. METHODS: Twenty one patients (10 women, 11 men, mean age 34 yr) had an enteroscopy from the terminal ileum to the ligament of Treitz during an ileocolectomy performed for CD. All patients were subsequently enrolled in a placebo-controlled trial of mesalazine for the prevention of early endoscopic relapse; 10 patients received placebo, and 11 received mesalazine (1.5 g/day) for 12 wk after surgery. At the end of this trial, they all had a colonoscopy with inspection of the anastomosis and the neoterminal ileum. RESULTS: POE was completed up to the angle of Treitz in all 21 patients. In 10/21 cases (47%), mild lesions were found distributed at random along the small intestine 30 cm beyond the resection margin. At colonoscopy performed 12 wk later, lesions were found in 11/21 cases (52%) between section margin and were estimated to be 25 cm over the anastomosis. Endoscopic recurrence occurred in 5/10 patients who received placebo and 6/11 patients who received mesalazine. Endoscopic recurrence occurred in 5/10 patients having lesions at POE and in 6/11 patients who had no lesions. There was no relationship between endoscopic recurrence at 12 wk and presence of lesions at initial POE, whatever the postoperative treatment. CONCLUSION: Endoscopic lesions let in place after "curative" surgery have no influence on early endoscopic anastomotic recurrences in CD. PMID- 7484998 TI - Role of antineutrophil cytoplasmic antibodies in an ethnically distinct population: Korean patients with ulcerative colitis. AB - OBJECTIVES: Studies have suggested that antineutrophil cytoplasmic antibodies (ANCA), known as a useful diagnostic marker in patients with ulcerative colitis (UC), may have a genetic basis, particularly in association with HLA class II genes. Because most studies examining the role of ANCA in UC have been performed in ethnically undefined populations, we have analyzed ANCA status in an ethnically distinct group of patients with UC. METHODS: Serum samples from 24 Korean patients with a known diagnosis of UC and 58 healthy Koreans were examined for the presence of ANCA, using a fixed neutrophil enzyme-linked immunosorbent assay. ANCA-binding patterns were examined by indirect immunofluorescence. RESULTS: The incidence of ANCA in 83.3% of Korean patients with UC was significantly higher than in controls (p < 0.0001). The mean binding level at a 1:100 dilution and the titer of ANCA were significantly higher in patients with UC than in controls. Among UC patients with ANCA, there was also a high incidence of perinuclear binding pattern. In contrast, there was no relationship between ANCA and age of patients, duration, activity, or extent of disease. CONCLUSIONS: High sensitivity and specificity of ANCA in an ethnically distinct group of patients with UC show that ANCA expression may not be ethnically determined, and they confirm the utility of ANCA as a useful diagnostic indicator of UC in an ethnically diverse groups of patients. PMID- 7484999 TI - Gastroduodenal involvement in staging of nodal non-Hodgkin lymphomas: a clinical and endoscopic prospective study of 235 patients. AB - OBJECTIVES: The gastrointestinal tract is often the site of involvement of non Hodgkin lymphomas (NHL). The aim of this endoscopic prospective study was to verify the prevalence of the gastroduodenal involvement in patients in staging for NHL and to assess its impact on the choice of therapeutic strategies. METHODS: Two hundred and thirty-five consecutive patients were included in the study. Upper gastrointestinal endoscopy was performed, and biopsy samples were taken on every mucosal lesion and on macroscopically illness-free duodenal and gastric mucosa. The samples were submitted to histological examination, and the clinical stage of NHL was reevaluated. RESULTS: Sixty-one of the 235 patients exhibited histological involvement of gastric (40), duodenal (7), or both (14) mucosae. Endoscopic lesions were recorded in 51 patients, but the involved mucosa appeared macroscopically normal in 10 patients (16.3%). In 13 patients, the gastroduodenal involvement modified the clinical stage from I and II to III, indicating a different therapeutic approach. No difference was detected in the frequency of gastrointestinal involvement among the high, intermediate, and low grades of lymphoma malignancies. Thirty-five positive patients underwent a further endoscopic examination after the chemotherapy treatment. Although clinical remission was expected in all cases, 42.8% of them subsequently exhibited NHL. CONCLUSIONS: Upper digestive endoscopy plus biopsy sampling plays a necessary diagnostic role, not only when major clinical signs (hemorrhage) are present, but also in earlier stages of NHL (I and II), when a reevaluation of the therapeutic strategy may be indicated. In stages III and IV of illness, it may also prove useful in evaluating the efficacy of chemotherapy. PMID- 7485000 TI - Confirmation of successful therapy of Helicobacter pylori infection: number and site of biopsies or a rapid urease test. AB - BACKGROUND: Although a number of tests have been described to detect the presence of Helicobacter pylori in biopsy specimens, studies of positive and negative value have largely been performed on untreated patients; testing the reliability of posttherapy has not been done. METHODS: We examined the value of the number and site of biopsies performed and the method used for specimen evaluation posttherapy. For postantimicrobial therapy of 141 patients with previously confirmed H. pylori infection, three biopsies were taken, two from the antrum and one from the corpus. Individual slides were coded, randomized, and interpreted blindly by two pathologists. Furthermore, in 143 patients, a biopsy specimen was taken from the antrum and was immediately inserted into the gel of the rapid urease test, and the results were compared with those obtained from histopathology obtained at the same time. RESULTS: In 71 patients, H. pylori therapy was unsuccessful; in 61 (86%), all three sites were positive. The highest yield with a single large cup biopsy specimen was 94%; the lowest was 91%. Two antral biopsies were negative in 4% [95% confidence interval (CI) = 1-12%]. The combination of a biopsy from the angulus incisura and one from the greater curvature of the corpus correctly identified all treatment failures (95% CI = 95 100%). The rapid urease test was false-negative in 5% (95% CI = 1-13%); there were no false-positives. CONCLUSION: Use of either the rapid urease test or two antral biopsies for evaluation of success of antimicrobial therapy for H. pylori infection will result in a false declaration of cure in at least 5% of cases. Three large cup gastric mucosal biopsies for histology are recommended for evaluation of the success of anti-H. pylori therapy. PMID- 7485001 TI - Evaluation of a new urease reagent strip for detection of Helicobacter pylori in gastric biopsy specimens. AB - OBJECTIVES: Determine sensitivity and specificity of a new urease reagent strip (URS) test for detection of Helicobacter pylori in gastric biopsy specimens. METHODS: Six paired biopsy specimens were obtained from the greater curvature of the distal antrum, the lesser curvature near the incisura, and the corpus along the greater curvature during 66 procedures on 59 patients with endoscopic findings of gastric antral mucosal erythema or erosions, or gastric or duodenal ulcers. One biopsy from each site was tested with the URS. The second was evaluated with histology. A final antral biopsy was evaluated with a urea/gel test. RESULTS: Twenty-eight of the 66 cases were histologically positive, with H. pylori observed in at least one of the three biopsy sites. The URS test correctly identified all 28. Of 193 individual biopsy specimens, 78 were positive for H. pylori. The URS correctly identified 77. Sensitivity was 0.99, specificity 0.95, positive predictive value 0.93, negative predictive value 0.99, and kappa 0.92. Average time to positive was 20 min. Twenty-seven antral biopsies were histologically positive for H. pylori. The URS test correctly identified all 27, whereas the urea/gel test correctly identified 21. For antral sites, URS sensitivity was 1.00 and specificity 0.95 versus urea/gel test sensitivity of 0.75 and specificity of 1.00. CONCLUSIONS: In this sample, the URS test is as accurate as histology in diagnosing H. pylori infections, and it provides results in less time and at a lower cost than histology. PMID- 7485002 TI - Prevalence of Helicobacter pylori infection in patients with large gastric folds: evaluation and follow-up with endoscopic ultrasound before and after antimicrobial therapy. AB - OBJECTIVES: Large gastric folds may result from infectious, inflammatory, neoplastic, vascular, and infiltrative disorders involving a part or the entire gastric wall. Helicobacter pylori infection of the gastric mucosa is associated with an active gastritis characterized by infiltration of the mucosa and submucosa with neutrophils, eosinophils, macrophages, and lymphocytes. The purposes of the study were: 1) to study patients with large gastric folds noted on computed tomography, upper gastrointestinal series, or endoscopy, with endoscopy and biopsies and endoscopic ultrasound to determine the prevalence of H. pylori infection and the location of the thickening within the gastric wall; 2) to reexamine H. pylori-infected patients with EUS after antimicrobial therapy to determine whether resolution of the wall thickening accompanied eradication of H. pylori and improvement of histological gastritis. METHODS: Thirty-two patients with thickened gastric folds were studied. Eighteen patients had H. pylori infection and were treated with amoxicillin 1 g b.i.d. and omeprazole 40 mg b.i.d. x 14 days. One month after antimicrobial therapy, patients were reexamined by EUS, and gastric biopsies were obtained. RESULTS: Eighteen of 32 patients had H. pylori infection and gastritis. In the H. pylori-infected patients with gastritis, EUS demonstrated diffuse thickening of the inner three layers (mucosa lumen interface, deep mucosa, submucosa) without thickening of the 4th and 5th layers of the gastric wall. After antimicrobial therapy and resolution of gastritis, EUS demonstrated concomitant resolution of this thickening and normalization of layers 1-3. CONCLUSIONS: H. pylori gastritis is a common cause of gastric wall thickening. EUS allows intrinsic localization of the gastric wall thickening in patients with large gastric folds and H. pylori infection and documents the resolution of this wall thickening upon eradication of H. pylori and resolution of gastritis. PMID- 7485003 TI - Sensitivity, specificity, and predictive value of fecal occult blood testing (Hemoccult II) for colorectal neoplasia in symptomatic patients: a prospective study with total colonoscopy. AB - OBJECTIVES: To evaluate the specificity, sensitivity, and positive predictive value of fecal occult blood testing (FOBT) in symptomatic patients, with colonoscopy as the gold standard, and to assess the usefulness of FOBT as an ancillary examination in symptomatic patients for whom total colonoscopy is indicated. METHODS: We studied 439 consecutive patients who underwent Hemoccult II testing before total colonoscopy. RESULTS: The sensitivity, specificity, and positive predictive value for colorectal neoplasia (cancer and adenomatous polyps) was 76.5%, 56.7%, and 27.6%, respectively. The sensitivity, specificity, and positive predictive value for colorectal cancer were 69.2%, 73.2%, and 7.3%, respectively. CONCLUSIONS: The sensitivity, specificity, and positive predictive value of FOBT are difficult to estimate from screening programs, because Hemoccult-negative individuals do not undergo examination of the colon, and many of the Hemoccult-positive cases do not undergo total colonoscopy. With total colonoscopy serving as the gold standard, FOBT does not appear to be of much value as an ancillary examination in patients with symptoms potentially attributable to the lower gastrointestinal tract. PMID- 7485004 TI - Hepatotoxicity from isoniazid and rifampin in inner-city AIDS patients. AB - OBJECTIVE: To determine the incidence of hepatotoxicity due to isoniazid and rifampin in inner-city patients with active tuberculosis. DESIGN: A hospital based review of 70 consecutive in-patients in a 770-bed, inner-city hospital. The patient population is primarily African-American and Hispanic. METHODS: Fifty eight men and 12 women were followed from 2-12 wk (median 4 wk). Patients had to be treated for at least 2 wk to be eligible for the study. Patients were excluded if they had been on any anti-tuberculous or any other hepatotoxic drug during the 2-month period before their hospitalization. Aminotransferases, alkaline phosphatase, bilirubin, and albumin were obtained at least every 2 wk. RESULTS: Hepatocellular toxicity, defined as AST and/or ALT greater than 200 IU/L, occurred in eight out of 70 (11.4%) patients. The mean age of these patients was 38.9 yr (22-58 yr). Patients with AIDS were significantly more likely to develop hepatotoxicity than those with any other risk factor (p < 0.01). CONCLUSIONS: Baseline aminotransferases followed by monitoring may be necessary in AIDS patients. PMID- 7485005 TI - Epidemiology of hepatitis D virus infection: changes in the last 14 years. AB - To calculate the prevalence of hepatitis D virus (HDV) superinfection, antibody to HDV (anti-HD) was tested on admission in 696 hepatitis B virus (HBV) chronic carriers diagnosed between 1979 and 1992. Anti-HD was positive in 67 patients (9.6%), and it was more frequently detected in i.v. drug abusers (IVDA) (59/74, 79.7%) than in non-IVDA (8/622, 1.3%) (p = 0.0001). The prevalence of anti-HD was higher in HBV chronic carriers diagnosed between 1979 and 1985 (33/219, 15.1%) than in those diagnosed from 1986 to 1992 (34/477, 7.1%) (p < 0.001). However, these figures were not different when IVDA were analyzed alone; 29/34 (85%) and 30/40 (75%) (p = 0.4) IVDA diagnosed in both time frames were anti-HD-positive. Four hundred and thirty anti-HD-negative HBV carriers were prospectively followed, and serial determinations of anti-HD were made. Seroconversion to anti HD was observed in only six patients (1.4%), and again the rate of seroconversion was higher in IVDA (5/10, 50%) than in non-IVDA (1/420, 0.2%) (p < 0.0001). These findings show that IVDA HBV chronic carriers are still an extremely high risk group for HDV superinfection in Spain and that this virus has little penetrance in other epidemiological categories of HBV carriers. PMID- 7485006 TI - Endoscopic esophageal manometry. AB - OBJECTIVE: We performed through-the-scope-manometry of the esophagus on 12 patients referred for esophageal symptoms. METHODS: A 3-lumen polyvinyl tube was passed through the biopsy channel of a standard video-endoscope. All patients underwent esophagogastroduodenoscopy with through-the-scope-manometry as well as a conventional laboratory-based manometric study; the sequence of the procedures was randomized. RESULTS: Mean lower esophageal sphincter pressure was 18 +/- 11 mm of mercury by both methods. In the lower esophagus, mean wave amplitude was 60 +/- 25 mm of mercury by through-the-scope manometry and 82 +/- 28 by laboratory testing. In the upper esophagus, mean wave amplitude was 50 +/- 26 mm of mercury by through-the-scope manometry and 63 +/- 20 by laboratory testing. Wave duration tended to be lower by through-the-scope manometry than by laboratory testing in the lower and upper esophagus. In nine patients with normal esophageal motility, 54% of swallows resulted in a peristaltic wave by the endoscopic study versus 100% for the laboratory test. CONCLUSION: Through-the-scope-manometry was able to accurately measure lower esophageal sphincter pressure compared with laboratory based manometry. Peristaltic wave amplitude by through-the-scope manometry was reduced compared with laboratory-based manometry, most likely because of the use of dry swallows. Through-the-scope-manometry has promise as a screening test for esophageal motility disorders. PMID- 7485008 TI - Relationship between directly measured portal pressure gradient and variceal hemorrhage. AB - OBJECTIVE: It is commonly believed that variceal hemorrhage in patients with cirrhosis and portal hypertension does not occur below a portal pressure gradient (PPG) of 12 mm Hg. The aim of this study was to assess the relationship between directly measured portal pressure gradient and variceal hemorrhage. METHODS: The procedure of insertion of the transjugular intrahepatic portosystemic stent-shunt (TIPSS) for variceal hemorrhage provides access to the portal vein and allows direct measurement of the portal pressure. Right atrial, inferior vena caval, and portal pressure were recorded, and the PPG was calculated (portal pressure inferior vena caval pressure) in 48 patients undergoing TIPSS for variceal hemorrhage. RESULTS: PPG was reduced from a mean of 21.4 (6.4) before TIPSS to 10.6 (3.1) mm Hg after the procedure. Seven patients (14.7%) had a baseline portal pressure gradient of < 12 mm Hg. CONCLUSION: The results of this study do not support the concept of a discrete bleeding threshold. PMID- 7485007 TI - Gastrokinetic effects of levosulpiride in dyspeptic patients with diabetic gastroparesis. AB - OBJECTIVE: Antidopaminergic drugs may be useful in diabetic gastroparesis because the inhibitory activity of hyperglycemia on gastric motility seems to be related to dopamine receptor stimulation. For this reason, we evaluated the effect of levosulpiride on gastric emptying, dyspeptic symptoms, and metabolic parameters of insulin-treated diabetic patients. METHODS: Under double-blind conditions, 40 longstanding, insulin-treated dyspeptic patients with autonomic neuropathy and delayed gastric emptying were randomly submitted, with an interval of 15 days, to 4 wk of administration of both levosulpiride 25 mg t.i.d. and placebo according to a cross-over design. At the beginning of the study and after levosulpiride or placebo treatment, the gastric emptying time of a standard meal was measured ultrasonically; gastrointestinal symptom scores and glycaemic control were also evaluated. RESULTS: Levosulpiride reduced significantly (p < 0.001) the gastric emptying time from 416 +/- 58 to 322 +/- 63 min, whereas placebo did not change it consistently (396 +/- 58 vs 372 +/- 72 min). Symptoms improved significantly (p < 0.001) with levosulpiride compared with placebo. However, there was no significant correlation between the acceleration of gastric emptying and the symptomatological improvement. The reduction of mean plasma glycosylated hemoglobin concentrations after levosulpiride (7.3 +/- 1.9 vs 5.8 +/- 1.3) was not significantly different (p = not significant) compared with placebo (6.8 +/- .7 vs 6.1 +/- 1.4). CONCLUSIONS: Our study first demonstrates that levosulpiride has an accelerating effect on the emptying of solids from the stomach of patients with diabetic gastroparesis. The drug is also effective in relieving upper gastrointestinal symptoms in patients whose gastric emptying times remain very slow. Our findings suggest, but do not prove, that better blood glucose control could be achieved with reduction of gastric emptying time; further trials are needed in this field. PMID- 7485009 TI - Does portal hypertension modify colonic mucosal vasculature? Quantification of alteration by image processing and topology. AB - OBJECTIVE: Endoscopic images of the colonic mucosal venous structure in eight cirrhotic patients with esophageal varices and in eight noncirrhotic subjects were analyzed to qualitatively determine whether or not the vasculature be altered by persisting portal hypertension. METHODS: On the endoscopic images, a venous network consisting of confluent venous branches draining into one penetrating vein was as a whole defined as one "venous unit." For each venous unit, the area of vein bed was calculated by means of an image processor. Each venous network was idealized to be a graphic representation consisting of lines (venous routes) and dots (origins, terminations, branching or anastomosing), on which two topological variables, the total number of zero-dimensional cells (a0) and the Betti number of dimension-1 (beta 1[K]), signifying the complexity and the number of venous anastomoses, respectively, were determined. RESULTS: The area of vein bed (mean +/- SD) was significantly greater (p < 0.01) for the cirrhotic patients (41 +/- 37 mm2) than for the noncirrhotic subjects (24 +/- 12 mm2). The a0 was also greater (p < 0.01) for the patients (68 +/- 29) than for the noncirrhotic subjects (42 +/- 17). The beta 1[K] for the patients was significantly greater (p < 0.01) for the cirrhotic subjects (7.1 +/- 4.4) than for the noncirrhotic subjects (2.6 +/- 2.9). CONCLUSION: The image processing and topological analysis of the colonic mucosal veins demonstrated that persistent portal hypertension alters the colonic mucosal venous structure by increasing the caliber of the veins and the number of merges (branching or anastomosing) between them. PMID- 7485010 TI - Serum levels of six pancreatic enzymes as related to the degree of renal dysfunction. AB - OBJECTIVES: Currently, serum total amylase, pancreatic isoamylase (P-amylase), lipase, trypsin(ogen), phospholipase A2 (PLA2), and elastase I are advocated to be useful in diagnosing pancreatic diseases. However, the most useful among the above six enzymes in patients with impaired renal function has not been fully clarified. We, therefore, studied the relation of the serum levels of the above enzymes and creatinine clearance (CrCl) in normal controls and patients with chronic renal insufficiency or failure. METHODS: PLA2 and elastase I were assayed by RIA, trypsin(ogen) by EIA and others by activity. Subjects were 24 healthy controls and 47 patients with impaired renal function and no apparent pancreatic diseases. RESULTS: 1) Elastase I was significantly elevated only in patients with a CrCl of 10 ml/min or less, whereas others were elevated already in patients with a CrCl below 40 ml/min; 2) in 12 patients with a CrCl between 13 and 39 ml/min, lipase tended to be less frequently raised than others, except elastase I, although the difference was statistically insignificant; 3) in 28 patients with a CrCl between 40 and 74 ml/min, lipase was less frequently elevated than others, except elastase I and PLA2; 4) in seven patients with a CrCl of 10 ml/min or below, elastase I tended to be less frequently elevated than others, although the difference was statistically insignificant; and 5) the degree of elevation was within 2.5 times the upper limits of reference values in all enzymes, except trypsin(ogen) (within 4.8 times). CONCLUSIONS: Elastase I was least vulnerable to impaired renal function followed by lipase. We, therefore, recommend combined assays of elastase I and lipase for detecting pancreatic diseases in patients with renal insufficiency. When cut-off levels are set at 2.5 times the upper limit of reference values, P-amylase or PLA2 can replace lipase. PMID- 7485011 TI - Physiological effect of cholecystokinin on gastric emptying of liquid in functional dyspepsia. AB - OBJECTIVE: Early satiety and postprandial epigastric fullness are common symptoms in functional dyspepsia. Cholecystokinin (CCK), a mediator of satiety in humans, may be responsible for these symptoms through an increased effect on delaying gastric emptying. METHODS: In five normal subjects and in five patients, gastric emptying of inert liquid mixed with technetium (Tc99m) was studied during i.v. perfusion of normal saline and of physiological concentrations of CCK octapeptide. RESULTS: Administration of CCK significantly delayed emptying of inert liquid in patients and in normal subjects, and the effect was of similar magnitude in the two groups: residual gastric volumes at 90 min increased from 9.9 +/- 6.1 to 32.1 +/- 6.2% (p < 0.025) in controls and from 9.8 +/- 4.4 to 32.2 +/- 4.7% (p < 0.005) in patients during saline infusion in comparison with CCK infusion; also, prolongation of half emptying time was not different between the two groups (19.4 +/- 1.9 min to 39.4 +/- 15.2 min in controls and 19.5 +/- 3.0 min to 31.4 +/- 7.9 min in patients). CONCLUSIONS: We conclude that CCK at physiological concentrations acts similarly in normal subjects and in patients with functional dyspepsia; this suggests that, if this hormone is normally released after a meal, a peripheral action of CCK through delayed gastric emptying is not responsible for increased postprandial satiety in functional dyspepsia. PMID- 7485012 TI - The frequency and influence of gallbladder varices on gallbladder functions in patients with portal hypertension. AB - Gallbladder varices have been reported in patients with portal hypertension. The exact frequency and significance of these collaterals in patients with cirrhotic and noncirrhotic portal hypertension is not known. One hundred and two patients with portal hypertension [38 with cirrhosis, 29 with noncirrhotic portal fibrosis (NCPF) and 35 with extrahepatic portal vein obstruction (EHPVO)] and 25 healthy controls were studied. Gallbladder varices were seen at ultrasound as tortuous, dilated vessels in the wall or in the bed of the gallbladder. In 35 patients (19 patients with and 16 without gallbladder varices) and in 10 healthy controls, gallbladder functions were studied by determining fasting volume (FV) and then residual volume (RV) every 10 min over 1 h after giving a liquid meal of 420 k.cal. Ejection fraction (EF) was computed as a percentage by the formula: FV- RV/FV x 100. Twenty four (24%) patients had gallbladder varices: Five (13%) with cirrhosis, seven (24%) with NCPF, and 12 (34%) with EHPVO. FV in EHPVO patients was seen significantly more than in cirrhotics (31.6 +/- 15.4 vs 19.3 +/- 6.0 ml, p < 0.05). The RV and EF were not different in the three groups of patients compared with the controls. The EF was similar in patients with or without gallbladder varices (63.3 +/- 10.2% vs 64.6 +/- 10.4%). Gallbladder varices are often seen in portal hypertension, more often in EHPVO patients, and these collaterals cause some gallbladder stasis but do not impede gallbladder function and hence seem unlikely to contribute to gallstone formation. PMID- 7485013 TI - The "pseudo-cholangiocarcinoma sign" in patients with cavernous transformation of the portal vein and its effect on the serum alkaline phosphatase and bilirubin levels. AB - OBJECTIVES: The goal of this study was to identify the underlying disorder responsible for portal venous thrombosis and cavernous transformation of the portal vein (CTPV). All patients with this finding underwent a thorough medical examination with intent to determine the cause and biochemical consequences of CTPV. METHODS: During an 8-yr period, a total of 1247 patients with clinical evidence of portal hypertension were examined using ultrasonography. Forty four of these 1247 patients were found to have CTPV. In each case, the finding of CTPV was confirmed by portography using either splenoportography or arterial portography, with digital subtraction angiography. These 44 patients were studied in an effort to determine the etiology of the cavernous transformation. In addition, the specific reason for the increased serum bilirubin and alkaline phosphatase levels in 35 of the 44 cases was evaluated by endoscopic retrograde cholangiopancreatography (ERCP) (34 patients), percutaneous transhepatic cholangiography (one patient), and by CT in 19. The surgical findings in 10 of these 44 patients, who ultimately underwent splenectomy and portal venous decompression for bleeding, were reviewed in light of the ultrasonographic, portographic, and ERCP findings in the same 10 patients. RESULTS: The underlying disorder responsible for cavernous transformation was found to be Behcet's disease in seven patients, chronic liver disease in four, congenital hepatic fibrosis in five, congenital protein C deficiency in one, and a prior abdominal operation for cholelithiasis in one patient. Despite a full clinical, radiological, hematological, and chemical evaluation, no etiology for CTPV was found in the remaining 26 patients. All of these later cases had no indication for liver biopsy or evidence for parenchymal liver disease. In these 26 patients, the serum levels of bilirubin and alkaline phosphatase ranged from mild to moderately increased compared with the moderately to markedly increased levels present in the 18 patients having an identifiable underlying liver disease. Irregular, undulating narrowing and nodular extrinsic defects, the so-called "pseudo-cholangiocarcinoma sign" was present in 33 of the 35 patients who underwent either ERCP or percutaneous transhepatic cholangiography. No such findings were observed in 10 control cirrhotic patients with portal hypertension but without CTPV, who also underwent ERCP. CONCLUSION: The results of this study indicate that mildly increased serum alkaline phosphatase and direct reacting bilirubin levels occur in cases with CTPV associated with a pseudo cholangiocarcinoma sign. Presumably, these enzyme elevations are a result of compression of the biliary tree by the venous collaterals that run along the extrahepatic biliary tree. None of the 33 cases with this sign had cholangiolar carcinoma. Thus, when a patient with splenomegaly but without documentable parenchymal liver disease demonstrates an increase in the serum direct reacting bilirubin and alkaline phosphatase levels, CTPV and the presence of large extrahepatic venous collaterals partially obstructing the biliary tree should be suspected. PMID- 7485014 TI - Modulatory impact of acid and pepsin on esophageal hydrophobicity in humans. AB - OBJECTIVES: The role of hydrophobicity in the pathophysiology of the gastrointestinal tract is well established as a protective mechanism against the impact of lumenal acid and pepsin. Hydrophobic properties of esophageal secretion in humans remain largely unknown. METHODS: We have studied, therefore, hydrophobicity by using fluorescence probe in human esophageal secretion, elaborated under the impact of saline followed by HCl, HCl/pepsin, and final saline. RESULTS: Basal hydrophobicity of human esophageal secretion, elaborated during mucosal exposure to saline, was 237 +/- 32. This value, however, declined 72% during mucosal exposure to HCl (66 +/- 14 vs 237 +/- 32; p < 0.001) and 87% during mucosal exposure to acid supplemented with pepsin (30 +/- 4 vs 237 +/- 32; p < 0.001). Moreover, hydrophobicity upon perfusion with HCl/pepsin was 55% lower than after perfusion with HCl alone (30 +/- 4 vs 66 +/- 14), although the result was insignificant. Substitution of saline for HCl/pepsin solution during the last perfusion period resulted in a partial recovery of hydrophobicity in esophageal secretion (131 +/- 30 vs 30 +/- 4; p < 0.001), although this value was lower than the basal hydrophobicity value (131 +/- 30 vs 237 +/- 32; p = 0.028). In addition, we continuously observed a significant shift in the fluorescence emission maximum from 508 +/- 6.4 to 486 +/- 0.9 (p < 0.001) during perfusion with starting saline, to 492 +/- 1.6 (p < 0.001) during exposure to HCl, to 493 +/- 1.1 (p < 0.001) during perfusion with HCl/pepsin, and to 488 +/- 0.9 (p < 0.001) during infusion of final saline. The maximum emission wavelength after esophageal exposure to initial saline also was significantly lower than the maximum emission upon perfusion with HCl (492 +/- 1.6 vs 486 +/- 0.9; p < 0.05) and HCl/pepsin (493 +/- 1.1 vs 486 +/- 0.9; p < 0.05). Although basal hydrophobicity in males was similar to corresponding values recorded in females, mucosal exposure to HCl (pH 2.1) resulted in an 84% decline in females but only 60% in males. Therefore, the hydrophobicity value in females during the perfusion period with HCl was 52% lower than in males (p = 0.129). CONCLUSIONS: Esophageal secretion exhibits its hydrophobic nature presumably through the presence of mucus components such as mucin and mucin-associated phospholipids. The inhibitory impact of HCl and HCl/pepsin solutions on esophageal hydrophobicity may play a role in the pathogenesis of mucosal damage by gastroesophagel refluxate. PMID- 7485015 TI - Intestinal calcium absorption as shown by stable strontium test in celiac disease before and after gluten-free diet. AB - OBJECTIVES: To investigate the effect of gluten-free diet on mineral and bone metabolism in women with celiac disease and, using the strontium test, to assess intestinal calcium absorption. METHODS: We studied body mass index, biochemical and bone mineral indices, strontium absorption test, and bone mineral density in 18 women (mean age 36.8 yr, range 18-68 yr) with celiac disease at diagnosis and after 12 months of gluten-free diet. RESULTS: Mean strontium absorption at diagnosis was markedly decreased with respect to control values (13.84 +/- 9.03% vs 22.47 +/- 4.21%, p < 0.0001), and 11 of the 18 patients (61%, subgroup A) had low values. In all patients, mean hemoglobin, serum potassium, magnesium, plasma calcium, urinary calcium, and phosphorus were significantly abnormal at diagnosis, whereas only the subgroup A had significantly reduced body mass index, 25 OH vitamin D, and elevated alkaline phosphatase. This subgroup differed in body mass index (p < 0.003) and calciuria (p < 0.035) with respect to the other patients. Strontium absorption correlated with body mass index, calcemia, and 25 OH vitamin D. After the gluten-free diet, all biochemical variables and strontium absorption normalized (23.23 +/- 5.54%), whereas bone mineral density did not change. CONCLUSIONS: At diagnosis, the patients frequently had intestinal calcium malabsorption, as demonstrated by strontium test, with an early renal compensatory mechanism. After the gluten-free diet, the normalization of calcium absorption and the decrease of mid-molecule parathyroid hormone suggested a normalization of mineral metabolism, although a positive effect on bone mineral density was not evident at that time. PMID- 7485016 TI - Combined endoscopic thermal electrocoagulation with high dose omeprazole therapy in complicated heterotopic gastric mucosa of the esophagus. AB - We report a case of a 38-yr-old man presenting with dysphagia due to stricturing of an isolated patch of heterotopic acid-secreting gastric mucosa ("inlet patch") in the proximal esophagus who was successfully treated with endoscopic thermal coagulation while receiving acid suppression with a proton pump inhibitor. Follow up endoscopy at 15 months after completion of therapy revealed repopulation of the heterotopic gastric mucosa with normal squamous epithelium and continued resolution of the patient's symptoms. The clinical significance of the inlet patch is discussed, and the role of mucosal ablation in treatment of this entity is described. PMID- 7485018 TI - Nonsteroidal anti-inflammatory drug-induced colonic strictures: two cases and literature review. AB - We report two patients with large bowel submucosal diaphragm disease associated with nonsteroidal anti-inflammatory drugs (slow release form of diclofenac and phenylbutazone) who were admitted in 1990 and 1991 because of iron deficiency. At colonoscopy, the lumen of the ascending colon was divided into compartments by multiple thin circumferential mucosal membranes. Barium enema showed two short strictures in one patient. Right hemicolectomy was carried out on one patient. The other patient was simply advised to discontinue taking diclofenac and remains well. Such lesions are rare (10 cases have been reported) and resemble those previously described in the small bowel. PMID- 7485017 TI - Upper gastrointestinal hemorrhage associated with cutaneous application of diclofenac gel. AB - We present four cases of upper gastrointestinal hemorrhage associated with the cutaneous application of diclofenac (Voltaren Emulgel). The patients' ages ranged from 24 to 85 yr. Three of the patients had used the medication three times daily for at least 2 wk before the onset of bleeding. In two cases, the treatment was indicated for backache which, retrospectively, was attributable to a peptic ulcer. In those two patients, the hemorrhage was massive, requiring blood transfusions. Because systemic absorption of nonsteroidal anti-inflammatory drugs from topical formulations has been documented, caution should be exercised when prescribing these formulations to patients with a history of peptic ulcer. PMID- 7485021 TI - Superior pancreaticoduodenal artery aneurysm rupture presenting with hemorrhage from the ampulla of Vater into the gastrointestinal tract. PMID- 7485019 TI - Endoscopic management of pancreatic duct leak complicated by retrogastric abscess. AB - A patient was diagnosed with a large retrogastric abscess associated with a postsurgical pancreatic duct leak refractory to somatostatin (Sandostatin) and external drains. She was managed entirely by endoscopic means consisting of transgastric abscess decompression, irrigation via a nasocystic tube, total parenteral nutrition, and a 7-Fr pancreatic duct stent to facilitate healing of the pancreatic duct leak. Thirty days after placement, the pancreatic duct stent was removed and pancreatogram revealed closure. Fifty-one days after transgastric decompression of the abscess, near complete closure of the cavity was accomplished and the patient returned to a regular diet. One year later, the patient continues to do well. PMID- 7485020 TI - Cirrhosis: a risk factor for cryptococcal peritonitis. AB - Cryptococcal peritonitis is usually associated with end-stage renal disease and peritoneal dialysis. Significant liver disease has not been well recognized as a risk factor for its development. We report two patients with cirrhosis who developed peritoneal infections with Cryptococcus neoformans. We also retrospectively review all cases of cryptococcal illness at the Ohio State University Medical Center from October 1990 to January 1994 and present a review of the literature regarding cryptococcal peritonitis associated with hepatic dysfunction. Cirrhotic patients with this entity present with subtle, nonspecific complaints resulting in delayed diagnoses, dissemination, and death. We suggest that clinicians maintain an increased awareness of this unusual but lethal entity in patients with liver impairment. Early and frequent abdominal paracenteses with bedside inoculations of fungal culture medium, India ink preparations, and serum cryptococcal antigen testing may hasten the diagnosis and institution of appropriate therapy. PMID- 7485022 TI - Sickle cell intrahepatic cholestasis: approach to a difficult problem. AB - Sickle cell intrahepatic cholestasis is a rare but potentially fatal complication of sickle cell disease. Its characteristic features include hepatomegaly, extreme total hyperbilirubinemia, coagulopathy, and acute liver failure. Although the pathophysiology is uncertain, most reports in the medical literature indicate that the prognosis is grim. The only effective therapy that has been reported in this setting is exchange transfusion. We describe two hemoglobin SS patients with sickle cell intrahepatic cholestasis. We conclude that exchange transfusion and supportive care aimed at correction of coagulopathy, stabilization of the acute liver disease, and perhaps most important, avoidance of surgical intervention are the keys to a successful outcome. PMID- 7485024 TI - Endoscopic management of spontaneous bile duct perforation and bile peritonitis complicating term pregnancy. PMID- 7485023 TI - HIV-enteropathy and bile acid malabsorption: response to cholestyramine. AB - Chronic diarrhea and weight loss are common in patients with AIDS. We report on an AIDS patient with chronic diarrhea, steatorrhea, and marked weight loss. A 75SeHCAT test demonstrated that the diarrhea was mainly due to bile acid malabsorption. Therapy with cholestyramine dramatically reduced bowel movements and led to significant reversal of weight loss. PMID- 7485025 TI - Fecal impaction as a cause of acute lower limb ischemia. AB - Acute lower extremity ischemia secondary to fecal impaction has not been previously reported. Herein, we report the case of an elderly man who was referred to our medical center with an acutely ischemic right lower extremity and a possible abdominal aortic aneurysm. The suspicious abdominal mass noted by his local physician was found to be a large fecal impaction of the rectosigmoid which, by direct pressure, was compressing and occluding the right common iliac artery. After disimpaction, there was complete resolution of the vascular symptoms and signs in the right lower extremity. PMID- 7485026 TI - Sarcoidosis of the duodenum presenting as dyspepsia. PMID- 7485027 TI - Iatrogenic incarceration of umbilical hernia in cirrhotic patients with ascites. PMID- 7485028 TI - Signet-ring carcinoma in Crohn's disease presenting as a neck mass. AB - We report a case of signet cell carcinoma of the colon in a patient with Crohn's disease. Signet cell carcinoma, also termed linitis plastica, is an unusual malignancy of the colon and is rarely associated with Crohn's disease. In general, metastatic disease is evident at the time of diagnosis, and the prognosis is uniformly poor. The radiographic, endoscopic, and histological features are discussed. PMID- 7485029 TI - Jejunal diverticulitis manifesting with abdominal wall abscess. AB - Jejunal diverticulosis is generally considered to be an innocuous condition, but serious complications can arise and lead to acute or chronic syndromes. In this report, we describe a case of jejunal diverticulitis presenting with an abdominal wall abscess. To our knowledge, this is the first documented case of jejunal diverticulitis complicated by fistula formation leading to the development of an abdominal wall abscess. Because jejunal diverticula generally localize at the mesenteric border and their perforation tends to result in intra-abdominal abscess formation, we speculate that the abdominal wall abscess described in our case here was due to adhesions of jejunal loops to the abdominal wall secondary to previous abdominal surgeries. PMID- 7485030 TI - Hereditary linitis plastica of the stomach. PMID- 7485032 TI - Treatments for hepatitis C: refinements in the recipe? PMID- 7485031 TI - Hemangioma of the small intestine: case report and literature review. AB - One case of small bowel polypoid hemangioma of the jejunum presenting as recurrent gastrointestinal blood loss is reported. The presenting symptoms and diagnostic methods are discussed. The importance of intraoperative enteroscopy in diagnosing and treating this condition is emphasized. PMID- 7485033 TI - ANCA in IBD: nice, but necessary? PMID- 7485034 TI - Sarcoidosis, the pancreas, and diabetes mellitus. PMID- 7485035 TI - Patient tolerance: an important factor of dissatisfaction for colonoscopy. PMID- 7485036 TI - Re: [13C]urea breath test. PMID- 7485037 TI - Helicobacter pylori: the first description. PMID- 7485039 TI - Type C chronic hepatitis complicated by B-cell lymphoma. PMID- 7485038 TI - Re: Gastrointestinal blood loss with low-dose aspirin. PMID- 7485041 TI - Esophageal perforation after ingestion of colon lavage solution. PMID- 7485040 TI - Nonalcoholic steatohepatitis and the "bright liver syndrome": should a recently expanded clinical entity be further expanded? PMID- 7485042 TI - Esophageal tuberculosis with fistula to mediastinum. PMID- 7485043 TI - Cholesterol embolization leading to small and large bowel infarction. PMID- 7485044 TI - Acid peptic disease in relation to Ramadan fasting: a preliminary endoscopic evaluation. PMID- 7485045 TI - Estrogen replacement therapy and progression of intimal-medial thickness in the carotid arteries of postmenopausal women. ACAPS Investigators. Asymptomatic Carotid Atherosclerosis Progression Study. AB - The effect of estrogen replacement therapy (ERT) on 3-year changes in carotid intimal-medial thickness (IMT) was explored using serial B-mode ultrasound measurements collected during 1989-1993 as part of the Asymptomatic Carotid Atherosclerotic Progression Study (ACAPS). Eligibility included increased IMT and elevated low density lipoprotein cholesterol. Of the 186 postmenopausal ACAPS women randomly assigned to receive either placebo or lovastatin, 34% reported use of ERT. Users tended to be younger than nonusers by an average of 3 years, to have more favorable high and low density lipoprotein cholesterol levels, and to be more likely to have had hysterectomies. Baseline blood pressure, body mass index, and cross-sectional IMT were similar among ERT users and nonusers. In the placebo group, IMT tended to progress among ERT nonusers but to regress among ERT users: Mean covariate-adjusted progression rates were 0.015 +/- 0.007 mm/year versus -0.012 +/- 0.012 mm/year, respectively (p = 0.05). This difference appeared to be independent of lipoprotein concentrations. Lovastatin was associated with an approximately 25% lowering of low density lipoprotein cholesterol among both ERT users and nonusers and had a marked impact on IMT progression (p = 0.004) in these women. ERT appeared to have little additional effect on IMT in women assigned to lovastatin. ERT may reduce or halt the progression of early atherosclerosis in women not receiving active lipid-lowering medication. PMID- 7485046 TI - Blood pressure reactivity to stress varies by hypertensive status and sex in Nigerians. AB - Previous studies among American and European populations have demonstrated larger blood pressure responses to behavioral challenges among men and hypertensive individuals. This is the first report of cardiovascular responses to behavioral challenges in a West African population. Blood pressure and heart rate changes in mirror image tracing and speech making tasks were recorded for 787 Nigerian civil servants participating in a comprehensive blood pressure survey conducted in Benin City, Nigeria, during 1992. Similar to findings in other populations, greater task-induced increases in systolic blood pressure (SBP) and diastolic blood pressure (DBP) were present among men than women (mean values of SBP = 22.1 vs. 18.3 mmHg, p < 0.001; and DBP means = 13.3 vs. 11.2 mmHg, p < 0.0001) and among hypertensives than normotensives (SBP means = 27.6 vs. 19.2, p < 0.0001; and DBP means = 14.1 vs. 12.1 mmHg, p < 0.05). An elevated prevalence of hypertension among men of higher staff status has been found in this population; however, higher staff status was not consistently related to cardiovascular reactivity, independent of hypertensive status. Additionally, hypertensive men who had speech-induced increases of SBP > 40 mmHg had significantly greater left ventricular mass index than did those hypertensive men with smaller SBP increases (p < 0.04). This study demonstrates that measures of cardiovascular reactivity to behavioral challenges have cross-cultural application, suggesting the need for further investigations of the interrelation of hypertension, cardiovascular reactivity, and left ventricular mass. PMID- 7485047 TI - Impact of father's education and parental smoking status on smoking behavior in young adults. The CARDIA study. Coronary Artery Risk Development in Young Adults. AB - Associations of parents' education and smoking with young adults' smoking were examined in participants aged 18-30 years at baseline (1985-1986) in the Coronary Artery Risk Development in Young Adults study. Significant (p < 0.05) inverse age adjusted associations of father's education with participant smoking status among black men, white men, and white women disappeared after adjustment for participant's education. Parental smoking status was directly related to participant smoking status for all race/sex groups. Participant education was strongly inversely related to participant smoking. Public health campaigns should consider influences of parental behaviors on children's behaviors and associations of limited education with adverse lifestyles. PMID- 7485049 TI - Unintentional weight loss in the United States. AB - Lower weight is usually considered advantageous to health, yet weight loss has been associated with increased mortality. An explanation for this paradox might be that the benefits of weight loss may depend on whether the loss is intentional or unintentional. The authors investigated whether intentional and unintentional weight loss differed in their associations with known risk factors for morbidity and mortality in a nationally representative sample of the US population. The sample consisted of 9,144 persons, aged 45 years and older, who answered questions regarding 1-year weight change in the diabetes risk factor Current Health Topic of the 1989 National Health Interview Survey (NHIS). Statistical analyses incorporated the sample weights and characteristics of the survey design. Relative to a common referent group, the factors associated with weight loss differed depending on whether the loss was defined as intentional loss, as unintentional loss, or regardless of intention. Restricting analysis to the 1,999 persons who lost weight, unintentional relative to intentional weight loss was significantly (p < 0.05) associated with older age, poorer health status, smoking, lower body mass index, and, in men only, widowhood and less education. Thus, unintentional weight loss may serve as a marker for factors that characterize persons at greater risk of mortality than persons undergoing intentional weight loss. Also, intention to lose weight may help clarify the relation between weight loss and mortality that, to this point, has shown counterintuitive results. Studies of the relation between weight loss and mortality should incorporate intention as a factor in the analysis. PMID- 7485048 TI - Association of the waist-to-hip ratio is different with wine than with beer or hard liquor consumption. Atherosclerosis Risk in Communities Study Investigators. AB - Specific alcoholic beverage associations with the waist-to-hip ratio were characterized in 12,145 African-American and white men and women ages 45-64 years. Estimated waist-to-hip ratios of those consuming more than six nonwine alcohol drinks/week and more than six wine drinks/week (vs. nondrinkers) were 0.007 larger (p < 0.001) and 0.009 smaller (p < 0.05), respectively. In similar comparisons, the odds ratios for a large waist-to-hip ratio were 1.4 (95% confidence interval 1.1-1.7) for nonwine and 0.45 (95% confidence interval 0.21 0.95) for wine intake. The opposite direction in adjusted associations for wine and nonwine (mainly beer) drinking supports the popular concept of the "beer belly." PMID- 7485050 TI - Capture-recapture and multiple-record systems estimation I: History and theoretical development. International Working Group for Disease Monitoring and Forecasting. AB - This paper reviews the historical background and the theoretical development of models for the analysis of data from capture-recapture or multiple-record systems for estimating the size of closed populations. The models and methods were originally developed for use in fisheries and wildlife biology and were later adapted for use in connection with human populations. Application to epidemiology came much later. The simplest capture-recapture model involves two lists or samples and has four key assumptions: that the population is closed, that individuals can be matched from capture to recapture, that capture in the second sample is independent of capture in the first sample, and that the capture probabilities are homogeneous across all individuals in the population. Log linear models provide a convenient representation for this basic capture recapture model and its extensions to K lists. The paper provides an overview for these models and illustrates how they allow for dependency among the lists and heterogeneity in the population. The use of log-linear models for estimation in the presence of both dependence and heterogeneity is illustrated on a four-list example involving ascertainment of diabetes using data gathered in 1988 from residents of Casale Monferrato, Italy. The final section of the paper discusses techniques for model selection in the context of models for estimating the size of populations. PMID- 7485051 TI - Capture-recapture and multiple-record systems estimation II: Applications in human diseases. International Working Group for Disease Monitoring and Forecasting. AB - This article evaluates the potential epidemiologic uses of capture-recapture, which include the primary area of determining disease frequency. Capture recapture may be a means to effectively "count" new cases (incidence) or count existing cases (prevalence). Specific applications of capture-recapture in epidemiology are presented, one of which is its use in estimating death rates in a region close to Calcutta, India. The method also has considerable potential to assess suicides, and it may be the only technique to assess disease frequency in developing countries. In addition to generating an estimate of population size, another application of capture-recapture is to assess the costs of ascertainment relative to the degree of accuracy. This approach provides a formal means for assessing the cost-benefits of lists for the identification of cases. The authors believe that with careful and appropriate use, capture-recapture methods will provide a new approach that can considerably improve our ability to monitor disease. PMID- 7485052 TI - Efficiency and accuracy of disease monitoring systems: application of capture recapture methods to injury monitoring. AB - Capture-recapture methods were employed to determine the most accurate and efficient approaches to monitor adolescent injuries. Multiple sources were used to ascertain cases of adolescent injuries that occurred between September 1 and December 31, 1991, in a single school district in metropolitan Pittsburgh, Pennsylvania. Eliminating the duplicate cases between the sources revealed 144 verified injuries; 127 (88.2%) were identified by student monthly recalls, 33 (22.9%) by daily attendance records, 58 (40.3%) by medical excuses, and 72 (50.0%) by a 4-month student recall. Capture-recapture analyses were undertaken to assess potential dependencies between the sources, to estimate the degree of underascertainment in the population, and to evaluate the efficiency of the individual sources and the combinations between them. It was estimated that 91% of the cases in the population were ascertained when all four methods of case finding were utilized. Furthermore, the analysis indicated that accurate injury estimates could be achieved using combinations of only two or three of the sources. An analysis of the efficiency of the methods of ascertainment revealed a trade-off between effort (the number of hours needed to identify cases) and the precision (coefficient of variation) of the injury estimates. Capture-recapture analysis not only provided an approach to evaluate and adjust for undercount but also offered a formal means to evaluate the most efficient combination of the sources to maximize completeness while minimizing effort. The use of these techniques has the potential to evaluate and improve injury surveillance as well as other disease monitoring systems. PMID- 7485054 TI - Social class and cancer patient survival in Finland. AB - The effect of social class on survival was assessed in a cohort of cancer patients identified from the nationwide population-based Finnish Cancer Registry. The cohort consisted of all reported cases of the 12 most common types of cancer occurring in Finland between 1971 and 1985 among persons born in 1906-1945 (n = 106,661). Social class information based on occupation was obtained individually for each patient from the population census of 1970. Both observed and corrected (i.e., cause of death-specific) 5-year survival rates were used in the analyses. A statistically significant linear effect of social class on age-adjusted relative risk of cancer death was observed in six of 12 cancer types among men and in nine of 12 among women; and the risk was highest for those in the lowest social class. The relative risk of death due to cancer for social class I (highest) relative to social class IV (lowest) was lowest in bladder cancer (relative risk (RR) = 0.46, 95% confidence interval (CI) 0.34-0.61) and kidney cancer (RR = 0.61, 95% CI 0.48-0.78) among men and in corpus uteri (RR = 0.51, 95% CI 0.36-0.72) and rectum cancer (RR = 0.56, 95% CI 0.42-0.74) among women. The differences between results obtained using corrected and observed survival rates were small. These findings indicate that social class is an important determinant of cancer patient survival. Additional research is required to clarify the etiology of the social class differences and to identify factors that could be used for developing strategies to diminish such inequalities. PMID- 7485053 TI - Maternal asthma and idiopathic preterm labor. AB - Previous studies suggest that women with asthma are at increased risk of preterm birth. Moreover, drugs (especially beta-agonists) used to treat asthma are also used to treat preterm labor. The authors carried out a case-control study of 555 women from three hospital centers with idiopathic preterm labor (< 37 weeks), including two overlapping (i.e., non-mutually exclusive) subsamples: cases with early idiopathic preterm labor (< 34 weeks) and cases with idiopathic recurrent preterm labor (< 37 weeks plus a previous history of preterm delivery or second trimester miscarriage). Controls were matched to cases according to race and smoking history prior to and during pregnancy. All subjects responded in person to questions about atopic, respiratory, obstetric, and sociodemographic histories. Subjects in the early and recurrent preterm labor subsamples were also asked to undergo spirometric testing with methacholine challenge 6-12 weeks after delivery. Cases were significantly more likely to report histories of asthma symptoms and physician-diagnosed asthma (matched odds ratios of 2-3) than controls, particularly those cases with recurrent preterm labor. No significant associations were observed, however, with methacholine responsiveness. These results could not be explained by residual confounding by smoking or other variables, nor by selective recall of asthma symptoms and histories by cases. Women with asthma are at increased risk of idiopathic preterm labor. The fact that no such association was seen with methacholine responsiveness suggests that nonatopic, noncholinergic mechanisms may link bronchial and uterine smooth muscle lability. PMID- 7485055 TI - Recall accuracy for prescription medications: self-report compared with database information. AB - A methodological study was performed in 1992 to evaluate the accuracy of self reported use of nonsteroidal antiinflammatory drugs (NSAIDs) and noncontraceptive estrogens that had been dispensed during the previous 12 years. A sample of 560 individuals dispensed NSAIDs or estrogens, and 140 individuals without NSAID/estrogen dispensations were selected from the Group Health Cooperative pharmacy database. Demographic, behavioral, and drug information was ascertained by telephone interview for 356 persons with and 98 persons without NSAID/estrogen dispensations. Of those with only a single NSAID dispensation, 41% (95% confidence interval (CI) 32-50%) were able to recall any NSAID use compared with 85% (95% CI 76-94%) for those with multiple NSAID dispensations. Thirty percent (95% CI 24-36%) recalled the NSAID name, and 15% (95% CI 10-20%) recalled both the name and dose. For estrogens, 78% (95% CI 70-86%) recalled the name, but only 26% (95% CI 17-34%) recalled the name and dose. Age, but not sex, appeared to influence recall accuracy: Persons 50-65 years of age recalled the NSAID name more accurately than those aged 66-80 (odds ratio (OR) = 1.8, 95% confidence interval (CI) 1.0-3.4). A similar advantage was noted for 50- to 65-year-old women in recalling the estrogen name (OR = 1.5, 95% CI 0.6-3.9). Drug name was recalled more frequently for exposures stopped 2-3 years prior to interview than for those stopped 7-11 years prior (OR = 3.0, 95% CI 1.6-5.7, and OR = 2.4, 95% CI 0.9-6.7, for NSAIDs and estrogens, respectively). Specificity was consistently high, ranging from 92% to 100%. This study suggests significant underascertainment of self-reported prescription drug exposure but little evidence that exposures are overreported. PMID- 7485056 TI - Assessing HIV vaccine effects. AB - It is unlikely that human immunodeficiency virus (HIV) vaccines will create impenetrable barriers to infection. When the barriers to infection are broken, however, vaccine effects on the progression of infection to disease and on the contagiousness of infection could be considerable. The usual outcomes of vaccine trials are either infection or disease. The authors argue that for HIV vaccines, the alternative outcome of contagiousness may be more important. Because of the long incubation period to acquired immunodeficiency syndrome (AIDS), a vaccine trial with AIDS as the outcome would be a long and costly undertaking. Because contagiousness is concentrated into the period of primary infection, vaccine trials assessing contagiousness would not take as long. An approach to assessing vaccine effects on the contagiousness of primary infection while simultaneously assessing protection against infection is presented. It involves randomizing vaccination of couples in whom both individuals are uninfected and one or both have a risk of infection outside the couple. In such a study, the vaccine effect on susceptibility to infection can be estimated from the proportions of vaccinated and unvaccinated couples in whom neither partner is infected. Estimation of the contagiousness effect also uses information on the frequency with which both partners are infected. In areas of the world where heterosexual epidemics are emerging within the context of concurrent partnerships, the randomization of vaccination of couples could increase the efficiency and decrease the costs of vaccine trials. PMID- 7485057 TI - Re: "Case-control study of residential radon and lung cancer in Winnipeg, Manitoba, Canada". PMID- 7485058 TI - Renal cell cancer in hypertensive patients more likely related to lead exposure than to antihypertensive medication. PMID- 7485059 TI - Causation and disease: a chronological journey. The Thomas Parran Lecture. 1978. PMID- 7485060 TI - Exploring the role of diet in modifying the effect of known disease determinants: application to risk factors of liver cirrhosis. AB - The analysis of the combined effects of nutritional factors with other putative disease determinants in log-linear or logistic models is methodologically complicated by the strong multicollinearity between nutritional factors, resulting in poor precision in estimating the parameters. Furthermore, the generally used multiplicative structure is not always the most appropriate for describing the resulting joint effect of two or more factors on the disease risk. The authors addressed such problems in a case-control study assessing the interactions between alcohol intake, chronic hepatitis C virus (HCV) infection, and nutrient intake on the risk of liver cirrhosis. During the period from November 1989 to May 1990, 282 patients admitted to the medical departments of the hospitals of the Province of L'Aquila (central Italy) were enrolled: 115 cirrhotic patients aged 24-82 years (78 of whom were males) hospitalized because of liver decompensation, and 167 control patients aged 25-84 years (100 of whom were males) admitted to the same hospitals for acute diseases unrelated to alcohol intake, infection with hepatotropic viruses, and nutrition. No dose effect relation was found between the intake of any nutrient and the risk of cirrhosis using classical methods. The analysis of principal components showed, however, that a pattern of higher lipid but lower protein and carbohydrate intakes was significantly associated with the risk of cirrhosis. The Breslow and Storer parametric family of relative risk functions showed that a multiplicative structure was the most adequate to describe the joint effect of nutritional pattern with alcohol intake and/or chronic HCV infection, whereas an additive structure best described the joint effect of chronic HCV infection and alcohol intake. In conclusion, the analysis of principal components and the Breslow and Storer family are useful tools to explore the role of diet on disease risk when precise pathogenic knowledge is not available. As an original finding, the authors suggest that a higher lipid intake, combined with lower protein and carbohydrate intakes, modifies multiplicatively the risk of cirrhosis associated with alcohol intake and/or chronic HCV infection. PMID- 7485061 TI - Physical exercise and reduced risk of nonfatal myocardial infarction. AB - While the inverse association between physical activity and coronary heart disease risk is well documented, questions remain regarding the intensity of exercise, the potential for confounding by other risk factors for coronary heart disease, and the role of blood lipids and apolipoproteins. The authors examined these issues in the Boston Area Health Study, a case-control study of 340 patients (266 men, 74 women) who survived a first myocardial infarction between January 1, 1982, and December 31, 1983, and 340 controls matched on sex, age, and residence. The relative risk of myocardial infarction for those in the highest quartile of physical activity, compared with the lowest, was 0.50 (95 percent confidence interval (CI) 0.31-0.80) for men and 1.00 (95 percent CI 0.41-2.43) for women. When subjects were categorized by level of energy expenditure on moderate to vigorous sports alone, men in the most active category had 0.39 (95 percent CI 0.23-0.69) times the risk of those in the least active category, and women, 0.43 (95 percent CI 0.15-1.26) times the risk. Adjustment for body mass index, smoking, alcohol intake, diet, personal and family medical history, and personality type did not substantially change results nor did further adjustment for blood lipids. This was not surprising as total energy expenditure was uncorrelated with blood lipids or apolipoproteins. Moderate to vigorous sporting activity, however, appeared to be directly related to high density lipoprotein (HDL) cholesterol (p = 0.06), especially the HDL2 subfraction (p = 0.10). In these data, findings suggest that physical activity is inversely related to myocardial infarction risk, independently of other risk factors for coronary heart disease. PMID- 7485062 TI - Urinary excretions of high molecular weight beta-thromboglobulin and albumin are independently associated with coronary heart disease in women, a nested case control study of middle-aged women in the Diagnostisch Onderzoek Mammacarcinoom (DOM) Cohort, Utrecht, Netherlands. AB - Increased plasma levels of beta-thromboglobulin, a platelet activation marker, are observed in coronary artery disease. Urinary albumin excretion, a marker of endothelial cell perturbation, is related to cardiovascular disease in diabetes. To test the value of these markers in predicting forthcoming coronary disease, the relations between urinary excretions of high molecular weight beta thromboglobulin (HMW-beta TGf) and albumin and subsequent coronary disease in a cohort of 15,484 middle-aged women were investigated in a nested case-control study. Baseline questionnaire data and urine samples were available from a breast cancer screening program in Utrecht during 1982-1985. Cases were Utrecht hospital admissions for myocardial infarction (n = 50) or angiographically confirmed coronary disease (n = 28) from 1982-1985 to 1990-1991. Classifying events occurred a median of 5.1 years after baseline. Controls were a random sample from the cohort, individually case matched for baseline examination date and age, at a 1:2 ratio. HMW-beta TG/creatinine ratios (ng/mmol, mean +/- standard error) were 5.3 +/- 0.3 in cases and 4.7 +/- 0.3 in controls; albumin/creatinine ratios (mg/mmol, median) were, respectively, 0.37 and 0.22. Crude odds ratios for classification in the highest compared with the lowest tertiles of HMW-beta TG/creatinine and albumin/creatinine distributions were elevated for cases compared with controls: HMW-beta TG/creatinine odds ratio = 2.4, 95% confidence interval 1.1-5.0; albumin/creatinine odds ratio = 2.1, 95% confidence interval 1.0-4.1. These relations persisted after adjustment for smoking, hypertension, Quetelet index, and menopausal status. Both urinary HMW-beta TG and albumin excretion are markers of coronary disease risk in middle-aged women, indicating that increased platelet activation and endothelial cell perturbation precede coronary heart disease events in women. PMID- 7485063 TI - Exposure to polychlorinated dioxins and furans (PCDD/F) and mortality in a cohort of workers from a herbicide-producing plant in Hamburg, Federal Republic of Germany. AB - The relation between mortality (all cause; cancer; cardiovascular diseases (International Classification of Diseases, Ninth Revision (ICD-9), codes 390 459); ischemic heart diseases (ICD-9 codes 410-414)) and exposure to polychlorinated dibenzo-p-dioxins and -furans (PCDD/F) was investigated in a retrospective cohort study. The cohort consisted of 1,189 male workers in a chemical plant in Hamburg, Federal Republic of Germany, who had produced phenoxy herbicides, chlorophenols, and other herbicides and insecticides known to be contaminated with 2,3,7,8-tetrachlorodibenzo-p-dioxin and other, higher chlorinated dioxins and furans. The authors reported previously on cancer mortality in this cohort for the follow-up period 1952-1989. The current study covers the years 1952-1992 and investigates the relation of PCDD/F exposure to mortality using a quantitative estimate of PCDD/F exposure for the whole cohort derived from blood and adipose tissue levels measured in a subgroup (n = 190). Quintiles and deciles of these estimates served as dose parameters in the estimation of relative risks (RRs), using year-of-birth stratified Cox regression. An unexposed cohort of gas workers served as an external reference group. The total mortality was elevated in all dose groups. The highest relative risk was observed for the highest 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD) decile (RR = 2.43, 95% confidence interval (CI) 1.80 to 3.29). Cancer mortality and mortality due to ischemic heart diseases showed a dose-dependent relation with TCDD and all PCDD/F combined. The highest relative risks for cancer (RR = 3.30, 95% CI 2.05 to 5.31) and ischemic heart diseases (RR = 2.48, 95% CI 1.32 to 4.66) were observed in the highest PCDD/F exposure group. The pattern of effects and tests for trend were similar when the lowest exposure group within the chemical worker cohort served as the reference, but the relative risks were smaller and the confidence intervals were larger. Potential confounding exposures complicate the interpretation of the internal comparison. These findings indicate a strong dose-dependent relation between mortality due to cancer or ischemic heart diseases and exposure to polychlorinated dioxins and furans. PMID- 7485064 TI - Psychologic stress in the workplace and spontaneous abortion. AB - The relation of work-related psychologic stress to spontaneous abortion was examined in a prospective study of 3,953 pregnant, employed members of a prepaid health plan in California, recruited between 1990 and 1991. Information on occupation, psychologic stress-related factors at work, social support at work, physical exertion at work, life events, pregnancy worries, and potential confounders was obtained by a telephone interview. Psychologic job stress and social support at work were assessed using an abbreviated version of the instrument developed by Karasek and colleagues (Los Angeles: University of Southern California, 1986). Results from the multivariate model indicated that, overall, stressful work was not associated with an increased risk of spontaneous abortion. However, interactions were observed between stressful work and maternal age over 32 years (p = 0.04), cigarette smoking (p = 0.02), and primigravidity (p = 0.06). Relative to the odds ratio for stressful work in young, nonsmoking, multigravid women without a history of two or more spontaneous abortions, the odds ratios for spontaneous abortion given stressful work were higher by 2.45 (95% confidence interval (CI) 1.03-5.81) for older women, 2.96 (95% CI 1.16-7.52) for smokers, and 2.27 (95% CI 0.97-5.27) for primigravid women. PMID- 7485065 TI - Epidemiology of preterm delivery in two birth cohorts with an interval of 20 years. AB - The occurrence and determinants of preterm delivery were studied in two population-based birth cohorts from northern Finland. In the first cohort of 1966 there were 11,475 singleton deliveries and in the later cohort of 1985-1986, 8,888. The overall incidence (percentage) of preterm deliveries fell from 9.1 to 4.8, including a reduction from 8.8 to 3.4 for spontaneous preterm deliveries. For iatrogenic ones, however, there was an increase from 0.3 to 1.4. The relative risks, associated with such known maternal determinants as unmarried status, smoking, low educational level, age over 34 years, unwantedness of the pregnancy, and poor earlier obstetric history, had about the same level in these two series. However, despite a favorable change in the distribution of most of these determinants over the 20 years, only a small part of the decrease of the total incidence of preterm delivery could be attributed to these changes, as a consistent and substantial reduction occurred in the incidence of spontaneous preterm birth in all categories of each single determinant. The increase in the proportion of iatrogenic preterm deliveries was accompanied by these being more common at lower than at higher socioeconomic levels in 1985-1986, whereas the social gradient appeared to be reversed in 1966. PMID- 7485066 TI - Bivariate logistic regression analysis of childhood psychopathology ratings using multiple informants. AB - A central issue in studies of risk factors for childhood psychopathology is utilization of the information obtained about the child's mental health status from multiple informants. In this paper, the authors propose a new approach to the analysis of risk factor data when the outcomes are binary ratings (presence/absence of symptoms). This new approach has several attractive features in this setting. The strategy taken is to perform a single analysis using multivariate modeling, in which simultaneous logistic regressions are conducted for the outcomes given by each of several informants. The advantages of this approach include the following: 1) it retains the complete information about case status for each informant; 2) it permits assessment of informant-risk factor interactions as well as "overall" risk factor effects; 3) it provides measures of association between the multiple informants and adjusts for the association between responses in the analysis; and 4) missing data on a subset of respondents can be incorporated in a straightforward way, permitting all subjects with at least one informant to be used in the analysis. To illustrate the methods, the authors present findings on risk factors for measures of "Internalizing" and "Externalizing" behaviors from two surveys using parent and teacher ratings of 6- to 11-year-old children in Connecticut between 1986 and 1989. PMID- 7485067 TI - Symptoms of dengue fever in relation to host immunologic response and virus serotype, Puerto Rico, 1990-1991. AB - The authors investigated the role of secondary immunologic response, virus serotype, age, and sex on the clinical manifestations of dengue fever in Puerto Rico. From surveillance data for 1990 and 1991, this study identified 3,926 laboratory-positive cases, including 889 for whom dengue immunologic status and symptoms could be ascertained. Of those, 622 cases were virologically confirmed, and 267 cases were serologically confirmed. More than 50% of all positive patients reported fever, chills, headache, eye pain, body pains, joint pains, nausea, vomiting, or skin rash. The frequency of reporting signs, symptoms, and hospitalization was significantly higher among persons with secondary infections diagnosed by serological methods. Only rash was more common among those with primary infections. Symptom reporting increased with age; body pains, joint pains, and rash were significantly more frequently reported by female patients. No significant difference in symptom frequency was found among the virologically confirmed cases, comparing primary and secondary cases or infections due to different serotypes. The data for serologically confirmed cases suggest that in Puerto Rico the manifestations of dengue fever are, as with dengue hemorrhagic fever in Asia, more prominent among those who are experiencing secondary infections, and this effect may be more marked in the younger age groups. PMID- 7485068 TI - Risk factors and cofactors for human T-cell lymphotropic virus type I (HTLV-I) associated myelopathy/tropical spastic paraparesis (HAM/TSP) in Jamaica. AB - Human T-cell lymphotropic virus type I (HTLV-I) has been etiologically associated with a neurologic syndrome called HTLV-I-associated myelopathy/tropical spastic paraparesis (HAM/TSP) as well as with adult T-cell leukemia/lymphoma. The authors sought to quantify the risk in Jamaica of HAM/TSP associated with HTLV-I infection and cofactors associated with this disease among infected individuals. Between 1988 and 1989, prevalent and incident HAM/TSP patients and controls with other neurologic diseases were enrolled in a retrospective study. A second control group was composed of HTLV-I-seropositive, asymptomatic carriers in Jamaica, ascertained in a separate study conducted in 1988. Although HTLV-I seropositivity was not a component of the case definition for HAM/TSP, all 43 HAM/TSP patients were HTLV-I seropositive compared with two (4.0%) of the controls with other neurologic diseases. Given HTLV-I seropositivity, one cofactor associated with the risk of HAM/TSP was young age at initial heterosexual confidence interval 1.29-12.46 for individuals aged < or = 15; odds ratio = 4.26, 95% confidence interval 1.41-12.90 for individuals aged 16-17 years at initial intercourse). Among individuals who reported this early age at initial sexual intercourse, an increased risk of HAM/TSP was associated with having reported more than five lifetime sexual partners (odds ratio = 2.88, 95% confidence interval 0.90-8.70). Neither an early age at initial sexual intercourse or the number of lifetime sexual partners was a risk factor for adult T-cell leukemia/lymphoma. These data support the hypothesis that HAM/TSP is associated with sexually acquired HTLV-I infection, whereas adult T-cell leukemia/lymphoma is not. PMID- 7485069 TI - Effect of older age on survival in human immunodeficiency virus (HIV) disease. AB - To evaluate the impact of older age (> 50 years old) on survival in late-stage human immunodeficiency virus (HIV) disease, the authors analyzed 846 HIV-infected patients at the San Francisco Veterans Affairs Medical Center from 1987 to 1992. The median age was 42 years with 171 (20.2%) subjects aged 50 or more years. Survival was measured from the date of initial lymphocyte testing (median CD4 count, 223 cells/mm3) until death or censoring. Compared with those aged less than 40 years, and after multivariate proportional hazards adjustment for other significant determinants of survival (CD4 percentage, CD8 count, hematocrit, and prior acquired immunodeficiency syndrome diagnosis), there was no difference in survival for those aged 40-49 years, but there was a trend toward decreased survival in those aged 50-59 years (relative hazard = 1.32, 95% confidence interval 0.90-1.94) and in those aged 60 or more years (relative hazard = 1.56, 95% confidence interval 0.99-2.46). The impact of older age on mortality in HIV disease is, however, less than the impact of age on overall mortality in the United States. Accordingly, while older HIV-infected patients do have a somewhat poorer survival, this risk need not be too highly emphasized in individual patients; older patients deserve aggressive management. PMID- 7485070 TI - Temporal change in the reproducibility of a self-administered food frequency questionnaire. AB - The authors studied temporal change in the reproducibility of a self-administered food frequency questionnaire. During 1988-1994, 492 residents of a rural Japanese town completed five questionnaires including 27 food items, with intervals ranging from 2 weeks through 5.5 years. The reproducibility decreased steadily over time for pairs of the questionnaires administered in the same season (median Spearman's r at 2 weeks and 5 years = 0.62 and 0.28, respectively). The reduction was less marked for those surveyed in different seasons (median r at 5 months and 5.5 years = 0.35 and 0.28, respectively). The short-term, different season correlation at 5 months was lower than the short-term, same season correlation at 1 year. For individual food items, a lower initial reproducibility, infrequent consumption, and a larger difference in seasonal intake were associated with a greater reduction in reproducibility over time. The results indicate that reproducibility studies should deliberately choose the intervals and the seasons for surveys. PMID- 7485071 TI - Alternative approaches for estimating prevalence in epidemiologic surveys with two waves of respondents. AB - Estimates of prevalence in epidemiologic surveys are prone to bias due to selective response. Therefore, much effort is devoted to reduce the number of nonrespondents. For example, individuals who do not respond in the first round of recruitment in mail surveys are usually contacted a second (or even third or fourth) time yielding consecutive waves of responses. Yet this sequence of waves is often neglected in epidemiologic analyses in that prevalence is simply estimated as the proportion of trait-positive individuals among the total group of respondents. This paper investigates alternative estimates of prevalence that might be used in surveys with two waves of respondents. The estimates are based on different assumptions on the relation of response rates with the trait of interest. As this relation is likely to vary from survey to survey depending on the specific circumstances under which the recruitment of participants is conducted, none of the estimates is universally preferable. The performance of the different estimates is assessed in a variety of hypothetical and empirical examples, and strategies are discussed to make the best use of the different estimates in the analysis of epidemiologic studies. PMID- 7485072 TI - Re: "Total cholesterol and high density lipoprotein cholesterol as important predictors of erectile dysfunction". PMID- 7485073 TI - Re: "Sexual transmission of human T-lymphotropic virus type I among female prostitutes and among patients with sexually transmitted diseases in Fukuoka, Kyushu, Japan". PMID- 7485074 TI - Re: "Depressive symptomatology in women and residential proximity to high-voltage transmission lines". PMID- 7485076 TI - Reticulated platelets in uremic patients: effect of hemodialysis and continuous ambulatory peritoneal dialysis. AB - Platelet RNA content can be detected by flow cytometry using thiazole orange staining to identify platelets recently released into the circulation. We studied platelet RNA content and platelet function in uremic patients under different treatment regimens. Four groups were studied: (I) 15 end-stage renal disease (ESRD) patients (10M/5F) on maintenance hemodialysis (HD); (II) 11 ESRD patients (6M/5F) on continuous ambulatory peritoneal dialysis (CAPD); (III) 8 patients with chronic renal failure managed conservatively (5M/3F); and (IV) 34 controls (20M/14F). A double color labeling technique using a phycoerythrin-tagged antibody against glycoprotein Ib (CD42b) and RNA labeling by thiazole orange was performed and read by flow cytometry. Aggregation studies were made in platelet rich plasma using ADP, epinephrine, collagen, arachidonic acid, and ristocetin. In group I, samples were also obtained after HD. Platelet counts did not differ among the groups. Aggregation studies showed a lower response to ADP and ristocetin in the HD patients, but not in the CAPD or in the chronic renal failure patients. The percentage of platelets with high RNA content in group I was significantly lower than in controls (3.72 +/- 1.72% vs. 9.05 +/- 3.53%, P < 0.01), but was also lower than in the remaining groups (I vs. II P < 0.01, and I vs. III P < 0.01). No differences were seen in platelet RNA content among groups II (8.67 +/- 2.73%), III (9.14 +/- 3.04%) and IV. In group I, the percentage of reticulated platelets decreased further after HD (2.14 +/- 1.09%, P < 0.01). Aggregation studies showed a significantly lower response to ADP and ristocetin in group I (P < 0.05), but not in groups II or III in comparison with controls. Aggregation response to ADP and ristocetin decreased after HD (P < 0.05). In conclusion, HD may decrease the percentage of RNA-rich platelets through elimination of the younger and more active platelets and worsen the thrombopathy present in uremic patients. PMID- 7485075 TI - Sustained activation of blood coagulation in patients with cerebral thrombosis. AB - Recent progress in the measurements of the hemostatic markers enables us to assess the detailed profiles of hemostatic activation in various diseases. To evaluate the degree of hemostatic system activation in patients with cerebral thrombosis, detailed coagulation studies were performed in 28 patients with acute phase cerebral thrombosis and in 36 with chronic-phase cerebral thrombosis, together with 6 with chronic-phase cerebral hemorrhage and 37 age-matched healthy volunteers. In both acute-phase and chronic-phase cerebral thrombosis, plasma levels of thrombin-antithrombin III complex, plasmin-alpha 2-plasmin inhibitor complex and D-dimer were significantly higher, and antithrombin III and protein C were significantly lower than those in the normal group. Plasma fibrinogen concentration was significantly higher in chronic-phase cerebral thrombosis than that in chronic-phase cerebral hemorrhage. No significant difference was found in these variables between acute-phase and chronic-phase cerebral thrombosis. In addition, there was no difference in these parameters between chronic phase cerebral hemorrhage and normal subjects. These findings indicate that a sustained activation of coagulation and fibrinolysis is present in cerebral thrombosis, and it might contribute to the pathogenesis of cerebral thrombosis. PMID- 7485078 TI - Platelet autoantibodies in patients with chronic liver disease. AB - The thrombocytopenia in chronic liver disease (CLD) has been attributed mainly to hypersplenism, although other factors such as reduced mean life span with increased platelet turnover have also been demonstrated. Immunological abnormalities have been described in the pathogenesis and progression of CLD. In this sense, many studies have reported elevated levels of platelet associated IgG (PAIgG) in patients with CLD, and it has been suggested that PAIgG could represent true antiplatelet antibody. In this study we used a glycoprotein (GP) specific immunoassay (MACE) to determine whether PAIgG or circulating antiplatelet antibodies, reacted against the GPIIb/IIIa or GPIb/IX complexes, in patients with CLD. Thirty-six patients with CLD of diverse etiology were studied (20 female, mean age 53 years, range 38-75 years). 23 out of 36 patients (64%) had anti-GP antibodies in MACE. Particularly, 12 had anti-GPIb, 4 anti GPIIb/IIIa, and 7 had both types of autoantibodies. The existence of these anti GP antibodies was not related with the blood platelet count or etiology of CLD. These data show that in patients with CLD of diverse origin, there is a high prevalence of autoantibodies reacting specifically with platelet membrane GP, which constitutes the first evidence of the specific nature of platelet-bound IgG in CLD. These findings suggest that in patients with CLD, an immune mechanism may participate in inducing or aggravating the thrombocytopenia. PMID- 7485079 TI - Danazol distribution in plasma and cell membranes as related to altered cell properties: implications for mechanism. AB - Concentrations of danazol in patient plasma and red blood cells (RBC) were assayed over a 6-month period in 75 patients on danazol therapy using a high pressure liquid chromatography (HPLC) method more reliable than previous radioimmunoassay (RIA) methods. It was found that plasma danazol rose regularly for 15 days after the beginning of treatment, reaching a steady state plateau of 175 +/- 76 ng/ml in 20 patients on normal dose, and less for lower dose schedules. After stopping danazol, concentrations declined to near zero in a similar time frame. RBC concentrations on a packed volume basis were similar to plasma levels. However, the membrane ghosts of RBC contained about 50% of the total RBC danazol, implying about 100-fold higher concentration in membranes than in plasma. Similar distributions were obtained in vitro with both RBC and platelets, and were confirmed by 14-C-labeled danazol. These findings tend to support the hypothesis that the benefits of danazol in immune disorders may be attributable in part to its intercalation in the lipid bilayer of the plasma membrane, altering antigen/receptor expression to modulate immune reactions. This hypothesis was first suggested when it was observed that the RBC of patients on danazol therapy showed morphological changes and increased resistance to osmotic lysis. It was later shown that danazol in vitro reduces binding of autoantibodies, and protects against complement-mediated lysis, suggesting direct action of danazol on the membranes. This hypothesis is discussed, and danazol's effect in protecting against complement-mediated lysis is described. PMID- 7485077 TI - Randomized trial of loperamide versus dose escalation of octreotide acetate for chemotherapy-induced diarrhea in bone marrow transplant and leukemia patients. AB - This study compares maximal daily doses of loperamide to escalating doses of continuous intravenous (CI) octreotide acetate in bone marrow transplant (BMT) and leukemia patients. Following chemotherapy, BMT and leukemia patients who developed > or = 600 ml of stool volume in a 24-hr period were randomized to receive loperamide 4 mg po q6h or octreotide 150 micrograms mixed in hyperalimentation solution or normal saline and administered CI. Patients were assessed at 48 hr intervals for decrease in stool volume from baseline. Complete response (CR) was defined as > or = 50% from baseline stool volume (BSV). Patients receiving octreotide who did not achieve a CR at 48 hr were dose escalated by doubling the dose to a maximum of 2,400 micrograms with evaluations at 48 hr intervals. Patients receiving loperamide who did not achieve a CR at 48 hr had treatment discontinued. A total of 36 patients were enrolled in the study. Of these, all were evaluable for intention to treat, and 31 were evaluable for initial response. Based on intent to treat at the initial 48 hr, patients receiving loperamide had a higher complete response rate (86% vs. 45%, P = 0.033) than did those who received octreotide. By treatment analysis (patients who actually received the drug), patients receiving loperamide had a higher complete response rate (92% vs. 56%, P = 0.0448) than did those who received octreotide at the 150 micrograms dosage level. Additional octreotide patients eventually achieved a CR at a higher dosage level (78%). Loperamide at maximal doses of 4 mg po q6h is more effective than octreotide 150 micrograms CI in treating diarrhea following chemotherapy in BMT and leukemia patients. Higher doses of octreotide may be required in a significant number of patients not responding to lower doses. PMID- 7485080 TI - Monoclonal antibodies in the management of acute leukemia. AB - This report reviews the diagnostic significance of immune markers, their relationship to patient outcome, and the therapeutic uses of monoclonal antibodies (MoAbs) in acute leukemia. Immunophenotyping allows for rapid and reproducible diagnosis in the majority of cases of acute leukemia. It is of particular importance in recognizing the major immunologic subclasses of acute lymphoblastic leukemia (ALL), and in identifying subtypes of acute myeloblastic leukemia (AML) which cannot be differentiated by morphology and cytochemistry alone, such as FAB M0 or M7. Immune marker analysis has been used to detect minimal residual disease in patients' bone marrow and CSF after treatment. However, the presence of leukemia-associated phenotypes on small numbers of normal cells may reduce the sensitivity of detection in some cases. The prognostic value of immune markers in AML is limited. In ALL, the prognostic significance of the different immunophenotypic subtypes has been lessened by modern treatment protocols. The relationship of mixed-lineage or biphenotypic antigen expression to patient outcome in both AML and ALL is unclear. Therapeutic applications of MoAbs in acute leukemia include immunologic techniques for purging malignant cells from autografts prior to transplantation, T-lymphocyte depletion from allografts as a strategy to reduce graft-versus-host disease, and the use of flow cytometry to monitor the timing and extent of leukapheresis in peripheral stem cell transplantation. MoAbs have also enabled the recent development of transplantation protocols using positively-selected CD34+ stem cells. PMID- 7485081 TI - Dysfibrinogenemia: a case with thrombosis (fibrinogen Richfield) and an overview of the clinical and laboratory spectrum. AB - Fibrinogen Richfield exemplifies a dysfibrinogen associated with a life-long thrombotic tendency. The evaluation of this novel case indicates that, like similar thrombotic dysfibrinogenemias, the abnormal protein polymerizes abnormally and demonstrates impaired clot dissolution. A survey of other cases of dysfibrinogenemia indicates that the relatively common abnormalities of Fibrinopeptide A release are generally asymptomatic or associated with bleeding, polymerization abnormalities are likely to be asymptomatic or associated with thrombosis (or occasionally bleeding), and complex abnormalities or additional, independent hemostatic defects are rather common. Thrombin and Reptilase clotting times are not helpful in distinguishing between the subsets, but clinical history, fibrinopeptide release, and polymerization studies may be useful. Abnormalities of fibrinogen function tend to correlate with changes in molecular domains related to binding and hydrolysis. PMID- 7485082 TI - CD2+, CD3-, CD56/NCAM+ malignant lymphoma with TCR beta gene rearrangement: a case report. AB - A case of CD56/NCAM+ malignant lymphoma is reported. Only a rare malignant lymphoma cell showed azurophilic granules in the cytoplasm of Giesma-stained preparations, while electron microscopic examination revealed occasional cytoplasmic granules with paracrystalline inclusions. The most common phenotype seen in NK lymphomas, CD2+, CD3-, CD56+, CD16-, CD57-, was present in the case. Cases with this phenotype have been interpreted to represent either true NK lymphoma or T-cell lymphoma with NK expression. Genotyping, where performed, has shown TCR germline configuration. Our case showed TCR beta rearrangement indicating that the above phenotype can be associated with a peripheral T-cell lymphoma. PMID- 7485083 TI - Two antithrombin mutations in a compound heterozygote: Met20Thr and Tyr166Cys. AB - The molecular basis for a family with Type I antithrombin deficiency has been established. Amplification and sequencing of the antithrombin gene identified two mutations: Met20Thr (2523T-->C) within exon 2 and Tyr166Cys (5493A-->G) within exon 3a. Further analysis indicated that the propositus was a compound heterozygote but in addition provided evidence for phase disruption during the amplification and/or cloning procedure. The Met20Thr mutation appears to be a neutral mutation with no functional consequences. In contrast, the Tyr166Cys mutation is associated with a Type I phenotype. PMID- 7485084 TI - Acquired von Willebrand's disease: a rare manifestation of postpartum thyroiditis. AB - This report describes the diagnosis of acquired type I von Willebrand disease in a 30-year-old woman (G5P5) who presented with complaints of excessive bleeding in the postpartum period. The patient's additional complaints of fatigue, depression, and inability to lose weight resulted in laboratory testing that indicated hypothyroidism due to thyroiditis. Clinical symptoms and laboratory tests for von Willebrand disease and hypothyroidism normalized with L-thyroxine replacement. Thyroiditis resulting in symptomatic hypothyroidism occurs in 2-4 per cent of postpartum women. The possibility of underlying hypothyroidism should be considered for those patients, especially if they are parous women, who appear to have an acquired bleeding disorder suggestive of von Willebrand disease. PMID- 7485085 TI - Human immunodeficiency virus rather than hepatitis C virus infection is relevant to the development of an anti-cardiolipin antibody. AB - We have investigated whether or not a relationship exists between anti cardiolipin antibody (aCL) positivity and human immunodeficiency virus type 1 (HIV) and/or hepatitis C virus (HCV), and we have attempted to clarify which virus has close association with the development of aCL. We found that aCL positivity in HIV-infected patients was significantly higher than in HCV-infected patients. Furthermore, HIV/HCV dual-infected patients exhibited a higher aCL positivity than patients infected by HCV alone. From these results, we conclude that HIV rather than HCV plays an important role in the development of aCL. PMID- 7485086 TI - Glibenclamide-induced photosensitivity in a diabetic patient with erythropoietic protoporphyria. PMID- 7485087 TI - Beraprost sodium for refractory thrombotic thrombocytopenic purpura. PMID- 7485088 TI - Functional specificity of anticoagulant autoantibodies defined by protein cofactors and Tween 20. PMID- 7485089 TI - Anticardiolipin antibodies in patients with angioimmunoblastic lymphadenopathy with dysproteinaemia. PMID- 7485090 TI - Pancytopenia secondary to parvovirus B19 infection in a child with acute lymphoblastic leukemia, mimicking leukemic relapse. PMID- 7485092 TI - Persistent parvovirus B19 infection and pure red cell aplasia in Waldenstrom's macroglobulinemia: successful treatment with high-dose intravenous immunoglobulin. PMID- 7485093 TI - Hemoglobin C/beta+ thalassemia and sarcoidosis. PMID- 7485091 TI - Successful treatment of sorivudine-related agranulocytosis with granulocyte colony stimulating factor. PMID- 7485094 TI - Lack of prophylactic anticoagulant therapy is not associated with clinical thrombotic complications in patients with hemophilia who undergo orthopedic surgical procedures. PMID- 7485095 TI - Serum hyaluronan is increased in malignant lymphoma. AB - The serum concentration of hyaluronan (HYA) was measured in 41 patients with malignant lymphoma, including 21 patients with non-Hodgkin's malignant lymphoma and 20 patients with Hodgkin's disease. Thirty-four patients were studied at diagnosis. The remaining 7 patients had relapsing or resistant disease. The patients were categorized into four stages according to conventional staging procedures. The median serum HYA concentration in patients with malignant lymphoma was significantly higher (median 40.7 ng/ml; 95% confidence limits 26.1 57.6 ng/ml) than in an age-matched healthy reference group (median 14.5 ng/ml, 95% confidence limits 11-19.4 ng/ml) (P = 0.00032). The highest serum HYA levels were found in patients with relapsing/resistant disease, all being in stages III and IV (median 181.5; range 11.9-500 ng/ml), as compared to previously untreated patients (median 29.8; range 9.1-108) (P = 0.0002) and controls (median 14.2; range 6.7-51.2). Decreased uptake and degradation of HYA owing to malignant transformation of lymphatic tissue is the most plausible explanation to these findings. PMID- 7485096 TI - Lymphocytosis of large granular lymphocytes in patients with Hodgkin's disease. AB - Clonal disorders of large granular lymphocytes (LGL) of either CD3- (NK cell) or CD 3+ (T-cell) phenotype have been described. B-cell malignancies such as hairy cell leukemia and non-Hodgkin's lymphoma have been observed in association with the T-cell type of LGL leukemia. Here we report the occurrence of LGL lymphocytosis in four patients with Hodgkin's disease. Immunophenotyping studies showed that these LGL were CD 3- in three patients and CD3+ in the other. LGL were polyclonally expanded in both patients in whom clonality could be assessed. PMID- 7485098 TI - Phenotypic and functional analysis of lymphocytes in paroxysmal nocturnal hemoglobinuria. AB - The hematologic disorder paroxysmal nocturnal hemoglobinuria (PNH) arises from a somatic mutation within the Piga gene important for the biosynthesis of glycosylphosphatidylinositol (GPI) anchors. The PNH defect has been identified in all cells of the myeloerythroid lineage, but involvement of the lymphoid lineage in PNH is more controversial. We therefore analyzed lymphocytes from 22 patients with PNH to characterize phenotypically the GPI-deficient population, and to investigate the functional consequences of GPI deficiency. GPI-deficient T lymphocytes, B lymphocytes, and NK cells were identified, but at a lower percentage than granulocytes and erythrocytes. CD8+ lymphocytes were significantly more affected than CD4+ T cells, and CD45RA+ lymphocytes were significantly more affected than CD45RO+ cells. Proliferation assays demonstrated that lymphocytes from PNH patients, either unfractionated or purified GPI deficient cells, responded normally to in vitro stimuli. When stimulated with phytohemagglutinin (PHA), naive CD45RA+ GPI-deficient T lymphocytes acquired the memory CD45RO+ phenotype. In addition, GPI-deficient T lymphocytes had a relative growth advantage as compared to normal T cells. The results demonstrate that PNH involves the lymphoid as well as the myeloerythroid lineage, and therefore arises from a totipotent bone marrow stem cell. The in vitro growth advantage of GPI deficient lymphocytes in PNH may have important implications for the pathogenesis of some puzzling clinical aspects of PNH, including predominance of the PNH clone, defective hematopoiesis, and leukemogenesis. PMID- 7485099 TI - Biphasic response of complement to heparin: fluid-phase generation of neoantigens in human serum and in a reconstituted alternative pathway amplification cycle. AB - We describe a study of the effects of heparin on complement activation through the use of assays for fragment C4d, fragment Bb, and the S-C5b-9 complex (S-MAC). In sera from healthy volunteers, virtually no change was observed in C4d either as a function of time or of heparin concentration, whereas changes in Bb and S MAC were biphasic. This observation was explored in greater detail in the heparin concentration range 0.001-5.0 u/ml (5 x 10(-3) to 25 micrograms/ml). For both Bb and S-MAC, a significant (P < 0.05) increase in production was noted in the heparin concentration range, 0.01-0.5 u/ml (5 x 10(-2) to 2.5 micrograms/ml). At higher heparin concentrations, Bb and S-MAC production decreased markedly (P < 0.05). We reconstituted the alternative pathway amplification cycle (C3, factor B, and factor D) and studied Bb generation. With reactants at concentrations one tenth those of normal serum, we observed a maximum generation of 13.2 micrograms/ml Bb. Control and heparin at 5 x 10(-4) micrograms/ml generated Bb concentrations of 6.8 and 6.1 micrograms/ml, respectively, for a 2-min incubation; at 5 x 10(-3) micrograms/ml heparin, Bb was increased to 9.8 micrograms/ml. Using isoelectric focusing to study anionic pI shifts in heparin bound factors B and D, it was found that factor B bound heparin only at the highest heparin concentration studied, i.e., 50 micrograms/ml; factor D, however, bound heparin at a much lower concentration (0.05 micrograms/ml). We conclude that, at low concentrations, heparin activates complement due to potentiation of the alternative pathway amplification cycle in the fluid phase. PMID- 7485097 TI - Hemostatic abnormalities and increased vascular endothelial cell markers in patients with red cell fragmentation syndrome induced by mitomycin C. AB - We examined red cell fragmentation syndrome (RCFS) induced by mitomycin C (MMC) (13 patients), by thrombotic thrombocytopenic purpura (TTP) (17 patients), and by disseminated intravascular coagulation (DIC) (15 patients). Plasma cytokine levels were increased in the TTP and DIC patients, but not in those whose RCFS was induced by MMC, suggesting that the activation of the immune system plays an important role in the pathogenesis of RCFS due to TTP and DIC but did not in RCFS due to MMC. Plasma thrombomodulin, tissue type plasminogen activator, and plasminogen activator inhibitor-I levels were increased in all RCFS patients, suggesting that RCFS, whether MMC induced, or due to TTP or DIC, might be associated with vascular endothelial cell injury. In TTP, von Willebrand factor (vWF) antigen and high molecular weight vWF multimer levels were reduced, possibly as a result of microthrombus consumption. The hemostatic data in this study showed that the TTP patients were in a hypercoagulable state without hyperfibrinolysis, and that DIC patients were in both a hypercoagulable and a hyperfibrinolytic state, whereas hemostatic abnormalities were slight in patients with MMC induced RCFS. These findings suggest that vascular endothelial cell injuries might be associated with RCFS, and that those injuries in MMC-induced RCFS might not be related to microthrombi or an activated immune system. PMID- 7485100 TI - Triosephosphate isomerase deficiency: repetitive occurrence of point mutation in amino acid 104 in multiple apparently unrelated families. AB - The molecular basis of triosephosphate isomerase (TPI) deficiency was studied in 3 patients from three separate families. In all 3 patients, genomic DNA directly sequenced after amplification by the polymerase chain reaction exhibited the point mutation TPI315C amino acid 104 Glu-->Asp. Although other mutations known to cause TPI deficiency have been restricted to single families, the amino acid 104 defect has now been described in nine apparently unrelated families throughout the world and is clearly the most frequently occurring form of the disorder. The basis of the repetitive occurrence of this mutation remains unexplained. PMID- 7485101 TI - Prophylaxis and therapy with factor VII concentrate (human) immuno, vapor heated in patients with congenital factor VII deficiency: a summary of case reports. AB - Hereditary factor VII deficiency is a rare autosomal recessive condition, usually associated with normal or reduced levels of a functionally defective molecule. The available means of treating this condition in North America presents serious health risks to the patient. Transfusion with fresh frozen plasma carries a risk of volume overload and a significant risk for viral transmission. Sustained prothrombin complex therapy is associated with a high risk for thrombogenic complications. This communication describes the use of Factor VII Concentrate (Human) Immuno, Vapor Heated--an intermediate purity factor VII concentrate from Immuno A.G.--for the treatment of 13 patients with factor VII deficiency. Treatment regimens described include those for long-term prophylaxis (three children), acute hemorrhages (two children, one adult), peripartum prophylaxis (one patient), and surgical coverage (two children, four adults). Prophylaxis and therapy were successful in all cases, the medication was well-tolerated, and there were no complications. In the three cases of long-term prophylaxis in children, doses of 10-50 IU/kg were given one to three times a week; one patient has undergone long-term prophylaxis for approximately 8 years, one patient for 1 year, and one patient for 1 1/2 years. Three cases in which Factor VII Concentrate was principally used for treatment of acute episodes of bleeding are described. One infant received Factor VII Concentrate on about 50 occasions for treatment of mucosal bleeding; a correction to 40-100% resulted in cessation of bleeding within 15 min in all cases. For treatment of an episode of intracranial bleeding, an 8-year-old boy received a dose of 37 IU/kg Factor VII Concentrate every 6 hr for peak factor VII levels of approximately 100% and troughs as low as 4% over the 11-day treatment period. A 37-year-old adult male with intracranial bleeding received alternating doses of 16 IU/kg and 8 IU/kg every 6 hr for 10 days with peak factor VII levels in the upper thirties (%). The peak favor VII level during surgical coverage with Factor VII Concentrate (neurosurgery, open reduction of ankle bones, dental surgery, pituitary adenoma surgery, closed liver biopsy) was approximately 100% in all cases, with trough levels ranging from 8 to 65% over treatment periods of 24 hr to 16 days using treatment intervals of 6-12 hr. PMID- 7485103 TI - A simple method for the derivation of the international normalized ratio for the reporting of prothrombin time results. AB - In order to harmonize the prothrombin time (PT) reporting systems, the World Health Organization, in collaboration with the International Committee on Thrombosis and Hemostasis, and the International Council on the Standardization in Hematology proposed a reporting system, the so-called international normalized ratio (INR). As the calculation of the INR depends on mathematical formula requiring a special calculator or mathematic tables, its derivation is associated with some difficulties and errors. In this article a nomogram is presented, by which derivation of the INR from different PT reporting methods can be easily done within a few seconds. PMID- 7485102 TI - In vitro and in vivo myelotoxicity of CAI to human and murine hematopoietic progenitor cells. AB - Carboxyamido-triazole (CAI), an agent that targets calcium-sensitive signal transduction pathways, has both antiproliferative and antimetastatic properties. The objective of this study was to evaluate the myelotoxicity of CAI to normal human and murine hematopoietic cells. In vitro toxicity of CAI was determined by inhibition of myeloid [colony-forming unit-granulocyte/macrophage (CFU-gm)] and erythroid [burst-forming unit-erythroid (BFU-e)] colony formation in clonal assays. The effects of oral CAI on CD2F1 mouse marrow and splenic cellularity, marrow progenitor content, and peripheral blood cell counts were assessed in relation to plasma CAI levels. In vitro, CAI caused a concentration-dependent inhibition of CFU-gm and BFU-e colonies following continuous drug exposure. Murine CFU-gm and BFU-e were inhibited > 90% by 10 and 15 micrograms/mL CAI, respectively. However, suppression of human CFU-gm and BFU-e did not exceed 65% at the same concentrations. In vivo, CAI reduced the number of CFU-gm and BFU-e per femur after the initial dose and through day 4. Variations in colony inhibition paralleled changes in CAI plasma concentrations. While colony inhibition increased in vitro with escalating drug concentrations, this was not observed in vivo with additional CAI doses. The low toxicity of CAI in vivo combined with the significant difference between toxicity for human and mouse progenitors in vitro suggests a relatively low adverse potential to the bone marrow for this new signal transduction inhibitory agent. PMID- 7485104 TI - Treatment of chronic neutropenia associated with large granular lymphocytosis with cyclosporine A and filgrastim. AB - A patient with neutropenia and life-threatening infections secondary to T-gamma lymphoproliferative disease, who did not respond to treatment with recombinant human G-CSF (filgrastim), was treated with filgrastin plus cyclosporine A (CyA). The patient achieved a good response in the absolute neutrophil count and subsequently required a dose reduction in the filgrastim. The patient was eventually discontinued from the CyA but continues on filgrastim alone. While on therapy, the large granular lymphocytes disappeared from the circulation and the beta-TCR rearrangement, which was present prior to beginning therapy, became undetectable. The patient had no significant toxicity to the CyA or the filgrastim and he has not experienced any serious infections or required hospitalization. Filgrastim has proven to be relatively nontoxic and of some benefit to patients with this disease and should probably be utilized first when treatment is necessary. However, if improvement is not observed, these findings suggest that a trial of the combination of CyA plus filgrastim may be beneficial. PMID- 7485105 TI - Heparin-induced thrombocytopenia and fatal thrombosis in a patient with activated protein C resistance. AB - We report a 20-year-old woman who developed heparin-induced thrombocytopenia (HIT) associated with devastating and fatal multiorgan thrombosis. The patient, her mother, and her brother were found to have resistance to activated protein C (APC), and the congenital thrombophilia in this family was verified by the finding of the Arg506 Gln mutation in factor V. This is the first case of HIT and APC resistance. The consequences of this association are discussed. PMID- 7485107 TI - Lymphoid malignancies following Kawasaki disease. AB - Kawasaki disease is a childhood multisystem vasculitic syndrome of unknown etiology. Associated immunological dysregulation is consistently observed and may be pathogenic in the known cardiac and noncardiac complications of the disease. Despite the well-known association between acquired immunodeficiency states and neoplasia, there are no reports of cancer accompanying or following the onset of this disease. We report here two children who were diagnosed with lymphoid malignancies shortly following the diagnosis of Kawasaki disease, suggesting an association between the two entities. PMID- 7485106 TI - Light-heavy chain deposition disease progressing to multiple myeloma. AB - Light chain deposition disease (LCDD) and light and heavy chain deposition disease (LHCDD) are rare clinical entities that have been associated with multiple myeloma, with monoclonal gammopathy of unknown significance (MGUS), or without any detectable protein abnormality. Renal failure is common, the diagnosis is difficult and prolonged survival is rare. The first patient with LHCDD and MGUS who progressed to multiple myeloma after 11 years is presented. A rising level of monoclonal IgA immunoglobulin, bone marrow plasmacytosis, and the presence of multiple bone marrow lesions on magnetic resonance imaging provided the first evidence of disease evolution. When management of serious complications permits a long survival, some patients with LCDD or LHCDD will develop multiple myeloma. PMID- 7485108 TI - Linkage-disequilibrium of the senegal haplotype with the beta S gene in the republic of guinea. AB - We have determined the beta gene cluster haplotype in 40 chromosomes carrying the beta S mutation from individuals of the major ethnic groups living in the Republic of Guinea. Thirty-one of these were either the typical (n = 29) or the atypical (n = 2) Senegal haplotype demonstrating that in this region of Atlantic West Africa, the sickle mutation is in linkage disequilibrium with this particular beta-gene cluster haplotype. Two individuals carrying one chromosome bearing the Benin haplotype were of Malinke ethnic origin. This is in keeping with the hypothesis that the Senegal beta-gene cluster haplotype is linked to the beta S gene in Atlantic West Africa. PMID- 7485109 TI - Improved survival of patients with chronic myelogenous leukemia undergoing allogeneic bone marrow transplantation. AB - A total of 28 patients with chronic myelogenous leukemia (CML) in chronic phase (CP) received bone marrow allografts from HLA-matched siblings at the University of Florida between August 1984-July 1992. The present study compares the disease free survival (DFS) for those patients who were transplanted before or after August 1988 using the same conditioning regimen. The analysis shows significant difference in 3-year DFS for those patients transplanted post- vs. pre-August 1988 (69.6% vs. 20%, respectively; P = 0.006). A decrease in pneumonitis due to different etiologies from pre-August 1988 (6/13, 46%) to post-August 1988 (1/15, 7%) was statistically significant (P = 0.029). A decrease, although statistically insignificant, in the overall incidence and severity of acute and chronic graft vs. host disease (GVHD) after August 1988 was also noticed. This study indicates significantly improved outcome for patients with CML in CP who have been treated in the University of Florida after August 1988. Better supportive care and prophylaxis for GVHD most likely contributed to such improvement. PMID- 7485110 TI - High-density lipoprotein fails to inhibit serotonin-induced activation of blood platelets. AB - High-density lipoprotein (HDL) of 100-400 micrograms/ml did not prevent morphological alterations of human blood platelets treated with serotonin (1-5 microM). Highly concentrated HDL (1,200 micrograms/ml) appeared to activate platelets in vitro. These findings indicate that whole HDL may not inhibit agonist-induced platelet activation. PMID- 7485111 TI - Rapidly progressive cutaneous vasculitis in a patient with chronic myelomonocytic leukemia. AB - This case report reviews the unique development of a vasculitic syndrome involving medium-sized arteries in a man with chronic myelomonocytic leukemia (CMMoL). This case has many features in common with cutaneous polyarteritis nodosa (CPAN), and this may represent the first instance in which CPAN developed in the setting of CMMoL. PMID- 7485113 TI - Intractable acute autoimmune hemolytic anemia in B-cell chronic lymphocytic leukemia successfully treated with vincristine-loaded platelet infusion. PMID- 7485114 TI - More on acute promyelocytic leukemia pneumonitis. PMID- 7485112 TI - Sustained granulocyte recovery after G-CSF in a patient with ticlopidine-induced severe aplastic anemia. PMID- 7485115 TI - N-Ras mutations: incidence and clinical relevance in myelodysplastic syndrome and acute myelogenous leukemia. PMID- 7485116 TI - Lymphoplasmocytic malignant lymphoma presenting with spinal epidural involvement: four reports of this uncommon lesion. PMID- 7485117 TI - Loss of HBsAg in chronic hepatitis B carrier following high-dose chemotherapy and autologous peripheral blood stem cell rescue for lymphoma. PMID- 7485118 TI - Analysis of chimerism by PCR using mutant-specific primers and sequencing of B gene region after allogenic bone marrow transplantation in sickle cell anemia. PMID- 7485120 TI - Allogeneic hemopoietic progenitor cells transplantation for late graft failure. PMID- 7485119 TI - Itraconazole and retinoid resistance. PMID- 7485121 TI - Donor-specific transfusion and donor bone marrow infusion in renal transplantation tolerance: a review of efficacy and mechanisms. AB - The ultimate goal in transplantation is modulation of the immune response to produce tolerance without immunosuppression. To date only a state of pseudotolerance for the allograft has been achieved through the use of potent pharmacologic and biologic manipulations. Despite these manipulations to prevent acute rejection, chronic rejection eventually results in graft failure. Thus, different strategies have been sought to induce tolerance and prevent acute and chronic rejection. Historically, donor-specific transfusion (DST) was one such strategy attempted. Donor-specific transfusion has been used primarily in living donor organ transplantation. With the concern that DST may sensitize patients, thereby preventing transplantation and introduction of cyclosporine, the use of DST was curtailed. More recently, donor bone marrow (DBM) infusion at the time of cadaveric transplantation has been used to facilitate development of microchimerism and tolerance to abrogate acute and chronic rejection. However, DBM infusion may predispose the recipient to graft-versus-host disease and is not easily accomplished in living donor organ transplantation. The potentially immunomodulating and tolerizing mechanisms of DST and DBM infusion are similar and include induction of anergy, stimulation of anti-anti-HLA antibodies, provision of soluble HLA antigen, suppressor cell and/or veto cell activity, clonal deletion, regulation of cell surface molecules, regulation of cytokines, promotion of microchimerism, or a combination of these. Of these mechanisms, microchimerism with the concomitant persistence of soluble donor HLA antigen is felt by many to be the most important. Although microchimerism is detectable in many patients who are tolerant of their grafts, there is no clear evidence that chimerism is responsible for the induction or maintenance of tolerance. PMID- 7485122 TI - Circadian blood pressure variation versus renal function. AB - Several published reports describe an abnormal circadian blood pressure profile in chronic renal failure subjects. Factors other than renal failure, including age, diagnosis of diabetes mellitus, autonomic dysfunction, and race, also may affect circadian blood pressure profiles. To further elucidate the relationship between renal function and circadian blood pressure variation, we compared day/night circadian blood pressure changes in three groups of male veteran hypertensive patients: group A, creatinine clearance (CC) > 80 mL/min, n = 20; group B, CC 20 to 80 mL/min, n = 19; and group C, CC < 20 mL/min, n = 14. We use postural changes in catecholamines, renin, and aldosterone as a measure of autonomic function. No significant difference in day/night percent change in systolic, diastolic, mean arterial pressure (MAP), or heart rate was seen by renal function group. Regression analysis using age, diagnosis of diabetes mellitus, postural hormonal changes, and creatinine clearance found race to be the only significant predictor of the day/night percent change in MAP (P < 0.05). Compared with whites, black subjects had higher nocturnal heart rates (P = 0.01); smaller day/night heart rate changes (P = 0.03); significantly higher diastolic blood pressure (P = 0.01); and a trend toward smaller day/night change in diastolic blood pressure (P = 0.06). In conclusion, renal function level does not influence day/night blood pressure changes. The blunting or reversal of the normal circadian blood pressure pattern seen in some chronic renal failure hypertensive subjects may be attributable to the association between chronic renal failure and cofactors associated with abnormal circadian blood pressure, including black race and possibly severity of atherosclerosis. PMID- 7485123 TI - Glomerular hyperfiltration in Koreans with non-insulin-dependent diabetes mellitus. AB - Glomerular hyperfiltration (GHF) may play an important role in the pathogenesis of diabetic nephropathy. The prevalence of GHF in non-insulin-dependent diabetes mellitus (NIDDM) is reported to vary in different populations, ranging from 35% in black Americans with new-onset diabetes to less than 5% in whites. We measured glomerular filtration rate (GFR) by 51Cr-EDTA clearance methods in 284 Korean NIDDM patients without overt nephropathy. Glomerular hyperfiltration (defined as GFR > 140 mL/min/1.73 m2) was found in 25% (71 of 284) of the patients. Thirty percent and 15% of the patients with known duration of diabetes less than 5 years and more than 10 years, respectively, showed GHF. Microalbuminuria (defined as urinary albumin excretion rate 20 to 200 micrograms/min) was found in 23% (64 of 284) of the patients. The mean GFR of the microalbuminuric patients was not significantly different from that of the normoalbuminuric patients (121.5 +/- 30.1 mL/min/1.73 m2 v 124.9 +/- 26.4 mL/min/1.73 m2; P = NS). However, when the patients with microalbuminuria were divided according to the presence or absence of retinopathy, the microalbuminuric patients with retinopathy had a lower GFR compared with the microalbuminuric patients without retinopathy (109.6 +/- 35.5 mL/min/1.73 m2 v 130.0 +/- 26.3 mL/min/1.73 m2; P < 0.01). These results show that the prevalence of GHF in Korean NIDDM patients without overt nephropathy is higher than that reported in whites and that it is similar to that in black Americans. The high prevalence of GHF in Korean NIDDM patients may contribute to the relatively higher prevalence of clinical diabetic nephropathy in this population. PMID- 7485124 TI - No association of converting enzyme insertion/deletion polymorphism with immunoglobulin A glomerulonephritis. AB - It has been recently reported that in type 1 diabetes the insertion/deletion (I/D) polymorphism of the angiotensin I-converting enzyme gene is associated with the presence of diabetic nephropathy. Tissue angiotensin I-converting enzyme is determined by I/D polymorphism, and it has been speculated that in diabetes differences of local angiotensin II availability determine the risk of renal disease. Since angiotensin II is thought to play an important role in the evolution of renal disease in general, we tested whether genotype distribution of the I/D polymorphism is also different in patients with immunoglobulin A glomerulonephritis (IgA-GN). Furthermore we compared IgA-GN patients with (1) stable renal function or (2) terminal renal failure to investigate a potential role of the I/D polymorphism in the renal prognosis. We examined 122 patients with biopsy-confirmed IgA-GN who had stable renal function and 82 dialysis dependent or transplanted patients with biopsy-confirmed IgA-GN. Furthermore, in 134 healthy individuals used as controls we analyzed the DNA for normal distribution of genotypes and allele frequencies. The polymorphic region was amplified using polymerase chain reaction with specific primers. Alleles were detected on 2% agarose gels. Genotype distributions and allele frequencies were not significantly different between controls and patients with IgA-GN and stable renal function. Furthermore, no significant difference in genotype distribution was detected between patients with IgA-GN and stable renal function compared with patients with IgA-GN and end-stage renal failure, although a trend for a higher frequency of DD-homozygotes was noted in the latter group (P = 0.07).(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7485125 TI - Splicing mutations in the COL4A5 gene in Alport's syndrome: different mRNA expression between leukocytes and fibroblasts. AB - The COL4A5 gene from 40 patients with Alport's syndrome was examined using single strand conformation substitution at the acceptor site (-2) of intron 50 and a G to-C substitution at the donor site (+1) of intron 47, respectively. The transcript in peripheral leukocytes from the former had a 10-nucleotide deletion. This shortened transcript was derived from abnormal splicing in a cryptic acceptor site within exon 51. This could be translated into a protein with an alteration of three amino acids followed by premature termination, which eliminated 23 amino acids from the carboxyl end. Gene tracking revealed that the mother and a brother carried the mutant allele. In the latter, the transcript in leukocytes was normal, but that in cultured skin fibroblasts showed skipping of exon 47, the result being that 71 amino acids were absent. Glomerular basement membrane from the patient did not react with the anti-alpha 5(IV) antibody. His maternal grandmother, mother, and a sister, all with abnormal urinalysis, carried the mutant allele. Thus, the appearance of exons of the COL4A5 gene in leukocytes may differ from that in fibroblasts. If kidney mRNA is not available, mRNAs from cultured skin fibroblasts, in addition to leukocytes, can be used for gene analysis in subjects with Alport's syndrome. PMID- 7485126 TI - Increasing incidence of focal-segmental glomerulosclerosis among adult nephropathies: a 20-year renal biopsy study. AB - Studies and textbooks from the 1970s and early 1980s list focal-segmental glomerulosclerosis (FSGS) as accounting for 10% to 15% of cases of idiopathic nephrotic syndrome in adults, although a recent review by D'Agati (Kidney Int 46:1223-1241, 1994) reported an approximately sevenfold increase in the incidence of FSGS from 1974 to 1993 in an active renal biopsy practice. To investigate possible changes in the incidence of FSGS in our renal biopsy practice, we reviewed reports from all nontransplant, adult (> or = 18 years) renal biopsies received in our laboratory from 1974 to 1993, which comprised 7,420 cases. All diagnoses of membranous nephropathy (MN), minimal change nephropathy (MCN), and FSGS made in each year were compiled; cases clearly or suspicious of being secondary to an underlying systemic disease, glomerulonephritis, or drug reaction were excluded. Relative frequencies of MN, MCN, and FSGS among these three diseases and among all biopsies were calculated for each year of the study. Regression analysis showed a significant (P < 0.001) increase in the odds of a diagnosis of FSGS over the study period: 7.6% per year among all biopsies and 6.8% per year among cases of MN, MCN, and FSGS only. Among all biopsies, the yearly incidence of FSGS increased from 4.0% +/- 0.6% (mean +/- SD) during the period between 1974 and 1979 to 12.2% +/- 2.0% during the period from 1987 to 1993. The odds of a diagnosis of MN (mean yearly incidence, 9.5% +/- 1.9%) did not vary significantly over the study period while the odds of a diagnosis of MCN (mean yearly incidence, 4.0% +/- 1.2%) declined at a rate of 2.2% per year (P < 0.03). Frequencies of diagnosis of MN, MCN, and FSGS by two pathologists were almost identical. Review of available slides from cases of FSGS revealed 21 (none before 1980) with characteristic histologic features of the collapsing glomerulopathy (CG) variant of FSGS. No more than four cases of CG were observed in any year of the study, and CG accounted for 4.7% of total FSGS cases for which diagnostic slides were available. Compared with 42 patients with non-CG FSGS, the CG cohort showed a greater percentage of black patients (86% v 38%), significantly higher mean levels of serum creatinine (3.8 +/- 2.7 mg/dL v 1.9 +/- 1.5 mg/dL) and urinary protein (14.3 +/- 9.6 g/24 hr v 7.7 +/- 5.8 g/24 hr) at the time of renal biopsy, and a greater likelihood of and more rapid progression to end-stage renal failure.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 7485127 TI - Sleep complaints are common in a dialysis unit. AB - Sleep disturbance among uremic patients is reported to be high, but data on the actual prevalence, clinical significance, and causative factors is limited. A sleep questionnaire was distributed to an entire hemodialysis unit of 64 patients. Of the 54 patients who completed the survey, 83.3% had sleep-wake complaints. Disturbed sleep was reported by 28 patients (51.8%), and causes were secondary to delayed sleep onset in 25 patients (46.3%), frequent awakening in 19 patients (35.2%), restless legs syndrome (RLS) in 18 patients (33.3%), and generalized restlessness in six patients (11.1%). Daytime sleepiness was the most frequent complaint, reported by 36 patients (66.7%), and RLS was the second most frequent specific complaint, reported by 31 patients (57.4%). Symptoms of sleep apnea were described by seven patients (13.0%). Male gender, age more than 60 years, RLS, and caffeine intake were associated with more sleep-wake complaints (P = 0.009, P = 0.002, P = 0.028, and P = 0.008, respectively). Urea and creatinine levels were higher in patients with RLS (P = 0.04 and P = 0.08, respectively); otherwise, no other metabolic or demographic variable was associated with specific sleep disorders or disturbance. Sleep problems are very common in dialysis patients and likely contribute to the impaired quality of life experienced by many of these patients. PMID- 7485128 TI - L-carnitine effects on anemia in hemodialyzed patients treated with erythropoietin. AB - To demonstrate whether L-carnitine treatment could further improve the anemia in dialyzed patients under recombinant human erythropoietin (r-HuEPO) therapy, leading to a reduction in r-HuEPO requirements, L-carnitine (1 g intravenously after every dialysis session) was administered for 6 months to a group of 13 patients; the results were compared with data from a placebo control group (N = 11). Globular osmotic fragility and endogenous EPO secretion were also evaluated. L-Carnitine treatment promoted a 38.1% reduction in r-HuEPO requirements in the active group (102.2 +/- 52.6 U/kg/wk v 63.3 +/- 37.8 U/kg/wk; P < 0.02), with globular osmotic fragility and endogenous EPO levels remaining unchanged and thus not accounting for carnitine effect on anemia. In the active group, seven patients decreased r-HuEPO needs (responders), while six did not (nonresponders). Compared with nonresponders, responders showed higher mean values at time 0 for r HuEPO requirements and endogenous plasma EPO levels, although not statistically significant. It is concluded that L-carnitine deficiency might promote EPO resistance in dialyzed patients, which is corrected by L-carnitine supplementation, ultimately reducing r-HuEPO requirements. PMID- 7485129 TI - Bacterial peritonitis in continuous ambulatory peritoneal dialysis: effect on dialysis on host defense mechanisms. AB - We have used an experimental model of continuous ambulatory peritoneal dialysis (CAPD) to investigate the effect of dialysis on host defense mechanisms in the peritoneal cavity. Laboratory rats undergoing regular dialysis were challenged with a single bolus of Staphylococcus epidermidis, Staphylococcus aureus, Candida albicans, and Pseudomonas aeruginosa. Intraperitoneal challenges with all four microbial strains underwent a rapid clearance during the first 4 hours, irrespective of whether the host was undergoing dialysis. Studies using S epidermidis showed that the rapid clearance was not mediated by the diaphragmatic stomata as might have been expected. Dialysis did, however, cause delays over the following 48 hours with all of the pathogens except C albicans. Bacterial clearance over a 10-day period was strain dependant, and microbial persistence was related to adhesion formation around the cannula. Again C albicans was an exception. Our conclusions are that dialysis does not affect the innate defense mechanisms involved in the early clearance of a bolus challenge to the peritoneal cavity but can cause delays in eradicating microorganisms that survive this phase. Because these experiments were carried out using a bolus inoculum, it may be necessary to reevaluate the effect of dialysis on host defenses in the peritoneal cavity under conditions in which the challenge more closely approximates a touch contamination event. PMID- 7485130 TI - Myeloperoxidase-antineutrophil cytoplasmic antibody-positive crescentic glomerulonephritis complicating the course of Graves' disease: report of three adult cases. AB - Antineutrophil cytoplasmic antibody (ANCA)-associated vasculitis has been recently recognized in Graves' disease patients treated with propylthiouracil. We have experienced three adult cases of Graves' disease with main features being renal derangements. All three patients, who were between the ages of 22 and 82 years, had been treated with propylthiouracil for 2 to 5 years after a diagnosis of Graves' disease. After several weeks of upper respiratory tract infection or flu-like symptoms, they abruptly began to manifest proteinuria and hematuria concomitant with severe anemia. Their serum creatinine increased from normal levels to 1.2 to 3.6 mg/dL. Renal biopsy revealed crescentic glomerulonephritis without deposition of immune complexes (ie, pauci-immune type). Crescent formations were observed in 40% to 60% of the glomeruli in all three cases. The serum from the patients revealed positive perinuclear-ANCA and negative cytoplasmic-ANCA (C-ANCA) pattern, and myeloperoxidase (MPO)-ANCA titers were 120 to 502 ELISA Units/mL (normal, < 10 ELISA Units/mL). A withdrawal of propylthiouracil with or without immunosuppressive therapy ameliorated their renal derangements. Graves' disease patients should be placed under vigilant observation by monitoring their urinalysis and serum creatinine, especially when being treated with antithyroid drugs and when suffering from flu-like symptoms. PMID- 7485131 TI - Successful use of recombinant human erythropoietin in a pregnant woman with lupus nephritis. AB - Recombinant human erythropoietin (r-HuEPO) is broadly accepted as treatment for anemia in dialysis and nondialysis patients with chronic renal failure, but data regarding the safety and efficacy of this drug in pregnancy are limited. Maternal and fetal problems have been reported to be associated with anemia during pregnancy. On the other hand, anemia is a frequent feature of systemic lupus erythematosus. We report the successful use of r-HuEPO in a young woman with lupus nephritis complicated by severe anemia during pregnancy. Additional studies should be encouraged to confirm the safety of r-HuEPO therapy during pregnancy. PMID- 7485132 TI - Hepatic infarction in a hemodialysis patient with systemic lupus erythematosus. AB - Hepatic infarction remains a rare and dramatic presentation of an acute abdomen in patients with systemic lupus erythematosus. It can be recognized preoperatively only with a high index of suspicion. Imaging studies such as abdominal computed tomography may aid in making the diagnosis. The tendency for thrombosis in these patients often can be associated with the presence of circulating anticoagulants from a group of antiphospholipid antibodies. The finding of lupus anticoagulant or anticardiolipin antibodies can help identify those patients more prone to thrombotic episodes, although at present it cannot be assumed that these antibodies cause the thromboses. Hemodialysis patients are said to have a decreased tendency for vascular thrombosis, but this may not be true in the presence of circulating antiphospholipid antibodies. We report a case of histologically documented hepatic infarction secondary to thrombosis in an end stage renal disease patient on hemodialysis with systemic lupus erythematosus and a circulating lupus anticoagulant. PMID- 7485133 TI - A patient with carpotarsal osteolysis and nephropathy treated with long-term hemodialysis. AB - We describe a type III carpotarsal osteolysis patient with nephropathy who has been maintained on chronic hemodialysis for 12 years. The diagnosis of osteolysis was based on x-ray findings and bone biopsy. At the age of 12 years, she developed renal disease that progressed; she was placed on chronic dialysis at the age of 18 years. Blood pressure was controlled by a medical regimen, and her anemia has benefitted from erythropoietin up to a level of 28% to 30%. Her osteolysis has stabilized since the age of 20 years; she is presently 30 years old. The major problem affecting the patient's clinical outcome is hyperphosphatemia. PMID- 7485134 TI - A randomized, placebo-controlled, multicenter trial evaluating alternate-day prednisone and fish oil supplements in young patients with immunoglobulin A nephropathy. Scientific Planning Committee of the IgA Nephropathy Study. PMID- 7485135 TI - Shaul G. Massry: a man of the world and a scholar for all seasons. PMID- 7485136 TI - A history of the parathyroid glands. AB - Diseases of the parathyroid glands have always existed, affecting occasional individuals in every period and culture of the past. The significant changes in body build associated with their dysfunction must have attracted in the popular mind bewildering connotations that can be gleaned from their mythology. Although their associated clinical features were described in the medical literature, the presence of the parathyroid glands and their function went long unrecognized. In the relatively slow evolution of endocrinology, compared with that of other medical disciplines, the parathyroid glands and their diseases were one of the last to be defined. It was only in the latter part of the nineteenth century that the glands were identified in humans by Ivar Sandstrom (1852-1889). Their subsequent experimental extirpation by Eugene Gley (1857-1930), as well as others, led to the differentiation of the neuromuscular symptoms that theretofore had been attributed to removal of the thyroid glands. That these symptoms were the result of the hypocalcemia that ensues parathyroidectomy was suggested in 1908 by W. G. MacCallum (1874-1944) and Carl Voegtlin (1879-1960). The possibility that parathyroid pathology may be associated with skeletal abnormalities was made in 1914 by Jacob Erdheim (18874-1937), and in the following year documented by Z. Schlagenhaufer from the observation that in patients with osteitis fibrosa cystica, only one parathyroid gland is enlarged. This was to set the stage for parathyroidectomy beginning in 1925, for the alleviation of bone disease of hyperparathyroidism.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7485137 TI - The role of the parathyroid glands in the uremic syndrome. AB - The genesis of hyperparathyroidism in uremia has turned out to be quite complex, involving low calcitriol, low ionized calcium, and possibly direct effects of high phosphate as well as the action of local factors and modifier genes determining the hyperplastic response of the gland. Growth is initially polyclonal and later monoclonal. In addition, the response of target tissues to parathyroid hormone (PTH) is attenuated ("PTH resistance"), and this may be due, at least in part, to diminished phenotypic expression of PTH receptors. In a series of elegant studies, it has been shown that PTH acts not only on the classical target organs of calcium homeostasis (ie, bone and kidney), but also on nonclassical. The role of PTH excess in the genesis of several features of the uremic syndrome, for example muscle dysfunction, cardiomyopathy, leukocyte and T cell dysfunction or insulin secretion by pancreatic islet cells, has been established. These studies have borne out the prediction that PTH is a "uremic toxin." PMID- 7485138 TI - Parathyroid hormone stimulates the generation of inositol 1,4,5-triphosphate in brain synaptosomes. AB - Parathyroid hormone (PTH) increases the levels of the second messenger, inositol 1,4,5 triphosphate (I1,4,5P3) in kidney and bone cells. It has been reported the I1,4,5P3 increases calcium uptake by brain synaptosomes. Because PTH also augments calcium entry in brain synaptosomes, it is possible that PTH induces the generation of I1,4,5P3 in these structures as well. The current study examined the effect of PTH-(1-84) on myoinositol turnover in vitro in rat brain synaptosomes. PTH-(1-84) in concentration of 10(-6)mol/L significantly (P < 0.01) increased the IP3 production (35 +/- 52%). The results indicate that PTH activates the phosphoinositol turnover in brain synaptosomes and that this pathway may be involved in the PTH-induced increase in [Ca2+]i in brain synaptosomes. PMID- 7485139 TI - Effects of recombinant human insulin-like growth factor 1 on renal handling of phosphorus, calcium, and sodium in normal humans. AB - The effects of insulin-like growth factor 1 (IGF-1) on renal handling of phosphorus, calcium, and sodium were evaluated in eight healthy men while they lived in a clinical research center for slightly more than 5 days. Subjects received a continuous intravenous infusion throughout the study of 0.45% saline and 2.5% d-glucose at 50 mL/hr (group 1, four subjects) or 150 mL/hr (group 2, four subjects). Recombinant human IGF-1 (rhIGF-1), 60 micrograms/kg body weight, was injected subcutaneously three times daily for 10 doses from day 2 until the beginning of day 5. After commencing the rhIGF-1 injections, there was a marked decrease in the fractional excretion of phosphorus in both groups that was sustained throughout the study. Urine phosphorus excretion also decreased significantly on days 2 to 5 in group 1 and on day 2 in group 2. In group 1, the fractional excretion of calcium decreased on day 2; urine calcium did not change. The fractional excretion of sodium decreased on days 2 and 4; urine sodium decreased significantly only on day 2. In group 2, the fractional and absolute urine excretion of calcium did not change. Fractional sodium excretion was not altered, and urine sodium increased only on day 5. The average serum phosphorus, calcium, and sodium did not change from baseline in groups 1 or 2. The pattern of the circadian rhythms for serum concentrations, urine excretion, and fractional excretion of phosphorus and calcium did not appear to be affected by the rhIGF-1 injections.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7485141 TI - Symptomatic hypercalcemia in a diabetic patient undergoing continuous ambulatory peritoneal dialysis: value of bone biopsy in the diagnosis and management. AB - An elderly man with diabetes mellitus and end-stage renal disease managed with continuous ambulatory peritoneal dialysis (CAPD) was hospitalized with peripheral vascular insufficiency; he developed hypercalcemia and became mentally obtunded. Lowering dialysate Ca from 3.5 mEq/L to 2.5 mEq/L, stopping calcium acetate, and ultimately hemodialysis with calcium-free dialysate did not lead to reversal of the hypercalcemia or improvement of his symptoms. The intact parathyroid hormone PTH level was 187 pg/mL, a value rarely associated with significant osteitis fibrosa. A search for other causes of hypercalcemia was unrevealing, and a iliac crest bone biopsy was done. The latter showed osteitis fibrosa, and the patient underwent parathyroidectomy. The hypercalcemia reversed quickly, and his mental symptoms slowly improved. The discussion reviews the probable causes of hypercalcemia in diabetic patient undergoing CAPD with 3.5 mEq/L dialysate calcium and using calcium-containing phosphate binders, with hyperparathyroidism certainly not the usual cause. The reason for the occurrence of significant hyperparathyroidism in the face of only modest elevation of PTH is considered. The value of bone biopsy in resolution of this problem is apparent. PMID- 7485142 TI - Relationship between intact 1-84 parathyroid hormone and bone histomorphometric parameters in dialysis patients without aluminum toxicity. AB - With the markedly reduced usage of aluminum salts in renal failure, parathyroid hormone (PTH) has become the major determinant of currently seen bone disease. Clinicians now must consider what PTH level should be sought. Too low a level may lead to the aplastic bone lesion (low turnover bone), and too high a level may cause osteitis fibrosa. Furthermore, conventional normal PTH levels may not be a suitable target because of the well-known resistance to PTH in uremic patients. In this report, we derive the PTH levels that best distinguish patients with low and high bone formation states from those with normal bone formation in a group of 175 dialysis patients without aluminum toxicity. Using bone histological parameters, we propose that ideally PTH levels should be maintained between 10 pmol/L (100 pg/mL) and 20 to 30 pmol/L (200 to 300 pg/mL) in chronic dialysis patients, levels two to four times the upper limit of values found in normal subjects. PMID- 7485140 TI - Phosphate transport inhibition by KW-3902, an adenosine A1 receptor antagonist, is mediated by cyclic adenosine monophosphate. AB - We have previously demonstrated that 1,3-dipropyl-8-(3-noradamantyl) xanthine (KW 3902) has an inhibitory effect on phosphate (Pi) transport with no effect on glucose transport in the rat renal proximal tubular cell, similar to that of parathyroid hormone (PTH). In the current studies we investigated the effect of KW-3902, rat PTH (1-34), and 1,3-dipropyl-8-cyclopentylxanthine (DPCPX), another selective adenosine A1 receptor antagonist, on Pi transport and the production of cyclic adenosine monophosphate (cAMP). We then compared these effects of KW-3902 with those of rat PTH in rat renal proximal tubule cells. The results showed that both KW-3902 (30 mumol/L) and rat PTH (1-34, 5 mumol/L) significantly inhibited Pi uptake in proximal cells from a control level of 61 +/- 3 to 19 +/- 3 (a reduction of 69%) and 46 +/- 4 picomoles phosphate/mg protein/min (a reduction of 25%), respectively (P < 0.01). The inhibitory effect of 30 mumol/L KW-3902 alone on Pi transport was more than twice that of 5 mumol/L rat PTH (1-34) alone (P < 0.01). KW-3902 stimulated the production of cAMP in a dose-dependent manner (r = 0.997, P < 0.01). Rat PTH (1-34; 5 mumol/L) also stimulated cAMP production, which was greater than that induced by 30 mumol/L KW-3902 alone. A significant increase in cAMP production by 30 mumol/L DPCPX was also observed.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7485143 TI - The importance of dosing intravenous calcitriol in dialysis patients with severe hyperparathyroidism. AB - The current study evaluates the use of intravenous (IV) calcitriol in 10 patients with severe hyperparathyroidism (HPTH). Patients with parathyroid hormone (PTH) > 1,200 pg/m and serum P < 6.5 mg/dL were studied. Ten patients with a mean PTH of 1,826 +/- 146 pg/mL were treated for a mean of 48 weeks with a dose of IV calcitriol commensurate to the level of PTH. The initial calcitriol dose had to be increased in seven patients. The mean maximum dose of calcitriol was 3.8 micrograms thrice weekly. There was a dramatic decrease in PTH levels, and by the end of the study it was 211 +/- 48 pg/mL. Alkaline phosphatase decreased from 582 +/- 3 to 120 +/- 12 IU/L. Serum Ca and P remained unchanged in most patients. There were three episodes of hyperphosphatemia in one patient, and another had a hypercalcemic episode. In conclusion, patients with severe HPTH respond very well to IV calcitriol, provided that dosing of calcitriol is commensurate to PTH levels, and hyperphosphatemia is kept under control. PMID- 7485144 TI - A new analog of calcitriol, 19-nor-1,25-(OH)2D2, suppresses parathyroid hormone secretion in uremic rats in the absence of hypercalcemia. AB - The active metabolite of vitamin D, calcitriol (1 alpha,25-(OH)2D3), suppresses parathyroid hormone (PTH) gene transcription. Although 1 alpha,25-(OH)2D3 is effective in suppressing secondary hyperparathyroidism (SH) in uremic patients, the mandatory use of large amounts of calcium salts to control serum phosphorus may preclude, in some patients, the use of ideal therapeutic doses of 1 alpha,25 (OH)2D3 because of hypercalcemia. We have studied a new analog of calcitriol, 19 nor-1 alpha,25-(OH)2D2, that possesses low calcemic and phosphatemic activity. Uremic rats received vehicle, 1 alpha,25-(OH)2D3 (2.0, 4.0, or 8.0 ng/rat) or 19 nor-1,25-(OH)2D2 (8.0, 25 or 75 ng/rat) intraperitoneally (IP) every other day for a period of 8 days. Pretreatment and posttreatment values of intact PTH were measured. The normal values for rat intact-PTH were 22 +/- 4.2 pg/mL and for ionized calcium (ICa) 4.77 +/- .07 mg/dL. The only dose of 1 alpha,25-(OH)2D3 that achieved a significantly, suppressed PTH (P < 0.01) was the 8.0 ng/rat. PTH decreased from 202 +/- 31 to 90 +/- 20 pg/mL. However, ICa increased from 4.81 +/ 0.08 to 5.08 mg/dL from uremic control (P < 0.02). Conversely, all doses of 19 nor-1,25-(OH)2D2 were effective in suppressing PTH, and none produced an elevation in ICa that was significantly different from that of vehicle-treated uremic rats. The maximum effect was achieved with the 75 ng/rat dose, which decreased PTH from 193 +/- 49 to 53 +/- 16 pg/mL (a decrease in 72.5%).(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7485145 TI - Renal afferent denervation prevents the progression of renal disease in the renal ablation model of chronic renal failure in the rat. AB - Rats with 5/6 nephrectomy develop hypertension and progressive deterioration in renal function. Several mechanisms may contribute to hypertension and to progressive renal disease in these rats. We have shown that increased activity of the sympathetic nervous system may contribute to hypertension in rats we chronic renal failure. However, the role of the sympathetic nervous system activity in the progression of renal disease has not been investigated. We have evaluated whether neurogenic factors contribute to the progression of renal disease in the renal ablation model of chronic renal failure in the rat. Sprague-Dawley rats underwent 5/6 nephrectomy and dorsal rhizotomy or sham rhizotomy (CRF). Age matched normal rats were used as controls. Six weeks after surgery, rats with chronic renal failure and dorsal rhizotomy had lower blood pressure and serum creatinine than rats with sham rhizotomy. In addition, kidneys from rats with 5/6 nephrectomy and rhizotomy manifested less severe glomerulosclerosis than kidneys from rats without dorsal rhizotomy. These studies have demonstrated in rats with renal ablation. Although normalization of blood pressure may definitely play a role, the data raise the possibility that neurogenic impulses to the kidneys may contribute to the progression of renal disease. PMID- 7485146 TI - Effects of pH on calcium transport in turtle bladder. AB - This study was designed to examine the effect of apical and basolateral (ie, mucosal and serosal) pH on calcium (Ca) transport in turtle bladder, a nonmammalian analog of the distal nephron. Unidirectional Ca45 fluxes were measured when serosal pH was 6.4, 7.4, or 8.4 (mucosal pH, 7.4) in the presence and absence of ouabain. When serosal pH was 8.4, M-->S Ca45 flux increased significantly, and when it was 6.4, M-->S Ca45 flux decreased markedly. Changes in serosal pH did not affect the S-->M Ca45 flux. When 5 x 10(-4) mol/L ouabain was added to inhibit sodium transport, M-->S Ca45 flux, at pH 7.4, was 221.6 +/- 27.4 pmol/mg/h (n = 10), and low pH again inhibited this flux (approximately 50%). Lowering mucosal pH (with serosal pH 7.4) also decreased M-->S Ca45 flux. In stripped bladders, Ca45 uptake increased linearly as medium pH was increased from 4.4 to 8.4. Total tissue Ca concentration did not change when serosal pH was varied, except at the extreme of pH 4.4, where tissue Ca decreased. By contrast, when apical pH was 6.4, tissue Ca rose substantially (approximately 1.5-fold). these results demonstrate that extracellular pH directly affects Ca homeostasis in the turtle bladder. Lowering the pH of either the serosal or mucosal medium directly inhibits apical Ca permeability. This change in Ca permeability is seen in the presence of ouabain. By contrast, alkalization of the serosal medium enhances apical permeability, but this effect is, in some manner, related to sodium transport.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7485147 TI - Use of magnetic resonance angiography to treat ischemic nephropathy. PMID- 7485148 TI - Four is not more than two. PMID- 7485149 TI - Prevalence of carriers of premutation-size alleles of the FMRI gene--and implications for the population genetics of the fragile X syndrome. AB - The fragile X syndrome is the second leading cause of mental retardation after Down syndrome. Fragile X premutations are not associated with any clinical phenotype but are at high risk of expanding to full mutations causing the disease when they are transmitted by a carrier woman. There is no reliable estimate of the prevalence of women who are carriers of fragile X premutations. We have screened 10,624 unselected women by Southern blot for the presence of FMR1 premutation alleles and have confirmed their size by PCR analysis. We found 41 carriers of alleles with 55-101 CGG repeats, a prevalence of 1/259 women (95% confidence interval 1/373-1/198). Thirty percent of these alleles carry an inferred haplotype that corresponds to the most frequent haplotype found in fragile X males and may indeed constitute premutations associated with a significant risk of expansion on transmission by carrier women. We identified another inferred haplotype that is rare in both normal and fragile X chromosomes but that is present on 13 (57%) of 23 chromosomes carrying FMR1 alleles with 53 64 CGG repeats. This suggests either (1) that this haplotype may be stable or (2) that the associated premutation-size alleles have not yet reached equilibrium in this population and that the incidence of fragile X syndrome may increase in the future. PMID- 7485150 TI - Somatic intragenic recombination within the mutated locus BLM can correct the high sister-chromatid exchange phenotype of Bloom syndrome cells. AB - Cells from persons with Bloom syndrome feature an elevated rate of sister chromatid exchange (SCE). However, in some affected persons a minority of blood lymphocytes have a normal SCE rate. Persons who inherit the Bloom syndrome gene BLM identical by descent from a common ancestor very rarely exhibit this high SCE/low-SCE mosaicism; conversely, mosaicism arises predominantly in persons who do not share a common ancestor. These population data suggested that most persons with Bloom syndrome in whom the exceptional low-SCE cells arise are not homozygous for a mutation at BLM but instead are compound heterozygotes. Following this clue, we carried out a genotype analysis of loci syntenic with BLM in 11 persons who exhibited mosaicism. In five of them, polymorphic loci distal to BLM that were heterozygous in their high-SCE cells had become homozygous in their low-SCE cells, whereas heterozygous loci proximal to BLM remained heterozygous. These observations are interpreted to mean that intragenic recombination between paternally derived and maternally derived mutated sites within BLM can generate a functionally wild-type gene and that low-SCE lymphocytes are progeny of a somatic cell in which such intragenic recombination had occurred. PMID- 7485151 TI - Mutations in SOX9, the gene responsible for Campomelic dysplasia and autosomal sex reversal. AB - Campomelic dysplasia (CD) is a skeletal malformation syndrome frequently accompanied by 46,XY sex reversal. A mutation-screening strategy using SSCP was employed to identify mutations in SOX9, the chromosome 17q24 gene responsible for CD and autosomal sex reversal in man. We have screened seven CD patients with no cytologically detectable chromosomal aberrations and two CD patients with chromosome 17 rearrangements for mutations in the entire open reading frame of SOX9. Five different mutations have been identified in six CD patients: two missense mutations in the SOX9 putative DNA binding domain (high mobility group, or HMG, box); three frameshift mutations and a splice-acceptor mutation. An identical frameshift mutation is found in two unrelated 46,XY patients, one exhibiting a male phenotype and the other displaying a female phenotype (XY sex reversal). All mutations found affect a single allele, which is consistent with a dominant mode of inheritance. No mutations were found in the SOX9 open reading frame of two patients with chromosome 17q rearrangements, suggesting that the translocations affect SOX9 expression. These findings are consistent with the hypothesis that CD results from haploinsufficiency of SOX9. PMID- 7485152 TI - Mutation T318M in the CYP11B2 gene encoding P450c11AS (aldosterone synthase) causes corticosterone methyl oxidase II deficiency. AB - Corticosterone methyl oxidase (CMO) deficiency refers to disorders of aldosterone synthesis due to mutations in the CYP11B2 gene encoding cytochrome P450c11AS, which is the adrenal aldosterone synthase. Type I CMO deficiency is associated with low concentrations of 18OH-corticosterone and aldosterone, due to severe mutations in P450c11AS; while type II CMO deficiency is associated with high concentrations of 18OH-corticosterone and low concentrations of aldosterone, due to less severe mutations of P450c11AS. A single type of mutation, compound homozygosity for R181W and V386A, has been reported as the cause of CMOII deficiency in an inbred population. We now report a patient with a typical clinical and hormonal picture of CMOII deficiency. Direct sequencing of patient and parent DNAs showed that the mother's allele contributed R181W and the deletion/frameshift mutation delta C372, while the father's allele contributed T318M and V386A. These mutants were recreated in cDNA expression vectors singly and in the parental pairs, showing that neither allele contributed any measurable activity. This would suggest the patient should have CMOI deficiency. These studies suggest that other factors besides P450c11AS are involved in the genesis of the distinctive CMOI and CMOII phenotypes. PMID- 7485153 TI - Molecular characterization of the breakpoints of a 12-kb deletion in the NF1 gene in a family showing germ-line mosaicism. AB - Neurofibromatosis type 1 (NF1) is caused by deletions, insertions, translocations, and point mutations in the NF1 gene, which spans 350 kb on the long arm of human chromosome 17. Although several point mutations have been described, large molecular abnormalities have rarely been characterized in detail. We describe here the molecular breakpoints of a 12-kb deletion of the NF1 gene, which is responsible for the NF1 phenotype in a kindred with two children affected because of germline mosaicism in the unaffected father, who has the mutation in 10% of his spermatozoa. The mutation spans introns 31-39, removing 12,021 nt and inserting 30 bp, of which 19 bp are a direct repetition of a sequence located in intron 31, just 4 bp before the 5' breakpoint. The 5' and 3' breakpoints contain the sequence TATTTTA, which could be involved in the generation of the deletion. The most plausible explanation for the mechanism involved in the generation of this 12-kb deletion is homologous/nonhomologous recombination. Since sperm of the father does not contain the corresponding insertion of the 12-kb deleted sequence, this deletion could have occurred within the NF1 chromosome through loop formation. RNA from lymphocytes of one of the NF1 patients showed similar levels of the mutated and normal transcripts, suggesting that the NF1-mRNA from mutations causing frame shifts of the reading frame or stop codons in this gene is not degraded during its processing. The mutation was not detected in fresh lymphocytes from the unaffected father by PCR analysis, supporting the case for true germ-line mosaicism. PMID- 7485154 TI - Molecular cloning of a full-length cDNA for dentatorubral-pallidoluysian atrophy and regional expressions of the expanded alleles in the CNS. AB - Dentatorubral-pallidoluysian atrophy (DRPLA) is an autosomal dominant neurodegenerative disorder characterized by genetic anticipation and variable combinations of symptoms including myoclonus, epilepsy, cerebellar ataxia, choreoathetosis, and dementia. Recently, we discovered that DRPLA is caused by unstable expansion of a CAG repeat of a gene on the short arm of chromosome 12. We determined the consensus DRPLA cDNA sequence containing the complete coding region for 1,185 amino acids. The CAG repeat, which is expanded in DRPLA, is located 1,462 bp downstream from the putative methionine initiation codon and encodes a poly-glutamine tract. Although poly-serine and proline tracts exist near the CAG repeats, these polyserine or proline tracts did not show any polymorphisms, which is in strong contrast to the high heterogeneity in the length of the CAG repeat. Northern blot analysis revealed a 4.7-kb transcript that is widely expressed in various tissues including heart, lung, kidney, placenta, skeletal muscle, and brain. Reverse transcription-PCR analysis revealed that the expanded alleles are transcribed to levels comparable to those of normal alleles. These results indicate that there is no difference in transcriptional efficiency between expanded and normal alleles. Furthermore, mRNA from cerebellar hemispheres of DRPLA patients showed smaller sizes of CAG repeats compared with other regions of the brain, which reflects somatic mosaicism of the expanded alleles of the DRPLA gene. PMID- 7485155 TI - The Friedreich ataxia critical region spans a 150-kb interval on chromosome 9q13. AB - By analysis of crossovers in key recombinant families and by homozygosity analysis of inbred families, the Friedreich ataxia (FRDA) locus was localized in a 300-kb interval between the X104 gene and the microsatellite marker FR8 (D9S888). By homology searches of the sequence databases, we identified X104 as the human tight junction protein ZO-2 gene. We generated a large-scale physical map of the FRDA region by pulsed-field gel electrophoresis analysis of genomic DNA and of three YAC clones derived from different libraries, and we constructed an uninterrupted cosmid contig spanning the FRDA locus. The cAMP-dependent protein kinase gamma-catalytic subunit gene was identified within the critical FRDA interval, but it was excluded as candidate because of its biological properties and because of lack of mutations in FRDA patients. Six new polymorphic markers were isolated between FR2 (D9S886) and FR8 (D9S888), which were used for homozygosity analysis in a family in which parents of an affected child are distantly related. An ancient recombination involving the centromeric FRDA flanking markers had been previously demonstrated in this family. Homozygosity analysis indicated that the FRDA gene is localized in the telomeric 150 kb of the FR2-FR8 interval. PMID- 7485156 TI - Molecular analysis of 24 Alagille syndrome families identifies a single submicroscopic deletion and further localizes the Alagille region within 20p12. AB - Alagille syndrome (AGS) is a clinically defined disorder characterized by cholestatic liver disease with bile duct paucity, peculiar facies, structural heart defects, vertebral anomalies, and ocular abnormalities. Multiple patients with various cytogenetic abnormalities involving 20p12 have been identified, allowing the assignment of AGS to this region. The presence of interstitial deletions of varying size led to the hypothesis that AGS is a contiguous gene deletion syndrome. This molecular analysis of cytogenetically normal AGS patients was performed in order to test this hypothesis and to refine the localization of the known AGS region. Investigation of inheritance of simple tandem repeat polymorphism alleles in 67 members of 24 cytogenetically normal Alagille families led to the identification of a single submicroscopic deletion. The deletion included loci D20S61, D20S41, D20S186, and D20S188 and presumably intervening uninformative loci D20S189 and D20S27. The six deleted loci are contained in a single YAC of 1.9 Mb. The additional finding of multiple unrelated probands who are heterozygous at each locus demonstrates that microdeletions at known loci within the AGS region are rare in cytogenetically normal patients with this disorder. This suggests that the majority of cases of AGS may be the result of a single gene defect rather than a contiguous gene deletion syndrome. PMID- 7485157 TI - Physical mapping of the holoprosencephaly critical region in 21q22.3, exclusion of SIM2 as a candidate gene for holoprosencephaly, and mapping of SIM2 to a region of chromosome 21 important for Down syndrome. AB - We set out to define the holoprosencephaly (HPE) critical region on chromosome 21 and also to determine whether there were human homologues of the Drosophila single-minded (sim) gene that might be involved in HPE. Analysis of somatic cell hybrid clones that contained rearranged chromosomes 21 from HPE patients defined the HPE minimal critical region in 21q22.3 as D21S113 to qter. We used established somatic cell hybrid mapping panels to map SIM2 to chromosome 21 within subbands q22.2-q22.3. Analysis of the HPE patient-derived somatic cell hybrids showed that SIM2 is not deleted in two of three patients and thus is not a likely candidate for HPE1, the HPE gene on chromosome 21. However, SIM2 does map within the Down syndrome critical region and thus is a candidate gene that might contribute to the Down syndrome phenotype. PMID- 7485158 TI - Physical mapping of the holoprosencephaly critical region in 18p11.3. AB - Holoprosencephaly (HPE) is a common developmental defect that results in a spectrum of craniofacial malformations. HPE is genetically heterogeneous, some cases being associated with deletions of the short arm of chromosome 18. In order to map the putative HPE gene located on 18p (HPE4) more precisely, six patients with various cytogenetic 18p deletions and clinical features of HPE have been characterized by using a combination of somatic cell hybrid analysis and FISH. By using a set of 27 chromosome 18p-specific markers, the deletion in each patient was characterized. The HPE minimal critical region on 18p was defined on a molecular level, localizing the HPE4 gene to 18p11.3. PMID- 7485159 TI - Congenital fibrosis of the extraocular muscles (autosomal dominant congenital external ophthalmoplegia): genetic homogeneity, linkage refinement, and physical mapping on chromosome 12. AB - Congenital fibrosis of the extraocular muscles (CFEOM) is an autosomal dominant syndrome of congenital external ophthalmoplegia and bilateral ptosis. We previously reported linkage of this disorder in two unrelated families to an 8-cM region near the centromere of human chromosome 12. We now present refinement of linkage in the original two families, linkage analysis of five additional families, and a physical map of the critical region for the CFEOM gene. In each of the seven families the disease gene is linked to the pericentromeric region of chromosome 12. D12S345, D12S59, D12S331, and D12S1048 do not recombine with the disease gene and have combined lod scores of 35.7, 35.6, 16.0, and 31.4, respectively. AFM136xf6 and AFMb320wd9 flank the CFEOM locus, defining a critical region of 3 cM spanning the centromere of chromosome 12. These data support the concept that this may be a genetically homogeneous disorder. We also describe the generation of a YAC contig encompassing the critical region of the CFEOM locus. This interval has been assigned cytogenetically to 12p11.2-q12 and spans the centromere of chromosome 12. These results provide the basis for further molecular analyses of the structure and organization of the CFEOM locus and will help in the identification of candidate genes. PMID- 7485160 TI - A gene that regulates DNA replication in response to DNA damage is located on human chromosome 4q. AB - Inhibition of replicative DNA synthesis following gamma-irradiation is observed in eukaryotic cells but is defective in cells derived from patients with the cancer-prone inherited disorder ataxia-telangiectasia (A-T) and in A-T-like Chinese hamster cell mutants. Chinese hamster cells show a less pronounced inhibition of DNA synthesis after gamma-irradiation when compared to irradiated human HeLa or mouse A9 cells. Therefore, to identify new human genes involved in the regulation of DNA replication in response to ionizing radiation in mammalian cells, single human chromosomes were introduced into Chinese hamster cells by microcell-mediated chromosome transfer. It is found that a new gene on human chromosome 4q inhibits DNA synthesis following gamma- and UV irradiation in hamster cells. However, this delay of DNA replication did not improve cell survival or the level of chromosomal aberrations induced by X-rays, indicating that the lack of the inhibition of DNA synthesis after X-irradiation is not a prerequisite for the X-ray sensitivity and chromosomal instability, which is observed in A-T and A-T-like hamster cells. PMID- 7485161 TI - Localization of a gene for autosomal dominant Larsen syndrome to chromosome region 3p21.1-14.1 in the proximity of, but distinct from, the COL7A1 locus. AB - Larsen syndrome (LS) is a skeletal dysplasia (osteochondrodysplasia) in which multiple dislocations of the large joints are the major feature. Nosology in this group of diseases, which constitutes 8% of Mendelian disorders in man, is primarily based on clinical and radiographic features. Hopes for more accurate classification grounds are currently being met by progress in elucidation of underlying genetic defects. We have performed linkage analysis in a large Swedish kindred with autosomal dominant LS and found the gene (LAR1) to be strongly linked to chromosome 3p markers (Zmax = 13.4 at (theta = .00). Recombination analysis indicates that the LAR1 locus is located in a region defined distally by D3S1581 and proximally by D3S1600, which cytogenetically maps to chromosome region 3p21.1-14.1. Linkage and recombination analysis of a COL7A1 PvuII intragenic polymorphism versus LS and chromosome 3 markers indicate that COL7A1 is located close to, but distinct from, the LAR1 locus. PMID- 7485162 TI - Mapping of a gene for long QT syndrome to chromosome 4q25-27. AB - Long QT syndrome (LQTS) is a heterogeneous inherited disorder causing syncope and sudden death from ventricular arrhythmias. A first locus for this disorder was mapped to chromosome 11p15.5. However, locus heterogeneity has been demonstrated in several families, and two other loci have recently been located on chromosomes 7q35-36 and 3p21-24. We used linkage analysis to map the locus in a 65-member family in which LQTS was associated with more marked sinus bradycardia than usual, leading to sinus node dysfunction. Linkage to chromosome 11p15.5, 7q35-36, or 3p21-24 was excluded. Positive linkage was obtained for markers located on chromosome 4q25-27. A maximal LOD score of 7.05 was found for marker D4S402. The identification of a fourth locus for LQTS confirms its genetic heterogeneity. Locus 4q25-27 is associated with a peculiar phenotype within the LQTS entity. PMID- 7485163 TI - Genetic homogeneity in Sjogren-Larsson syndrome: linkage to chromosome 17p in families of different non-Swedish ethnic origins. AB - Sjogren-Larsson syndrome (SLS) is a rare, autosomal recessive disorder that is characterized by congenital ichthyosis, mental retardation, and spastic diplegia or tetraplegia. Three United States families, three Egyptian families, and one Israeli Arab family were investigated for linkage of the SLS gene to a region of chromosome 17. Pairwise and multipoint linkage analysis with nine markers mapped the SLS gene to the same region of the genome as that reported in Swedish SLS pedigrees. Examination of recombinants by haplotype analysis showed that the gene lies in the region containing the markers D17S953, D17S805, D17S689, and D17S842. D17S805 is pericentromeric on 17p. Patients in two consanguineous Egyptian families were homozygous at the nine marker loci tested, and another patient from a third family was homozygous for eight of the nine, suggesting that within each of these families the region of chromosome 17 carrying the SLS gene is identical by descent. Linkage of the SLS gene to chromosome 17p in families of Arabic, mixed European, Native American, and Swedish descent provides evidence for a single SLS locus and should prove useful for diagnosis and carrier detection in worldwide cases. PMID- 7485164 TI - Evidence for an association between nonsyndromic cleft lip with or without cleft palate and a gene located on the long arm of chromosome 4. AB - Recent studies suggest that the familial aggregation of nonsyndromic cleft lip with or without cleft palate (CL +/- P) is likely to be attributable to the effects of several susceptibility loci, acting in a multiplicative fashion. Two potential CL +/- P susceptibility loci (CSL), transforming growth factor alpha (TGFA) and retinoic acid receptor (RARA), have been identified through association studies. In addition, recent evidence of linkage between CL +/- P and two markers (D4S175 and D4S192) in the region 4q25-4q31.3 raised the possibility that a CSL, with a larger effect than either TGFA or RARA, may reside within this region of the human genome. The present analyses were undertaken to determine whether D4S175 or D4S192 is significantly associated with CL +/- P in a sample of unrelated patients that have previously provided evidence of associations between CL +/- P and both TGFA and RARA. The results of these analyses provide further, tentative, evidence for the presence of a CSL locus on the long arm of chromosome 4 and help to refine the location of this locus in the region of D4S175 and D4S192. PMID- 7485165 TI - Mechanisms of small ring formation suggested by the molecular characterization of two small accessory ring chromosomes derived from chromosome 4. AB - Molecular cloning of a microdissected small accessary ring chromosome 4 from a moderately retarded and dysmorphic patient has been performed to identify the origin of the ring chromosome. FISH was performed with cosmids identified with the cloned, microdissected products and with other markers from chromosome 4. The present study clearly demonstrates that the small ring in this patient originates from three discontinuous regions of chromosome 4: 4p13 or 14, the centromere, and 4q31. It is suggested that the origin of the ring chromosome is a ring involving the entire chromosome 4, which has then been involved in breakage and fusion events, as a consequence of DNA replication generating interlocked rings. A second severely retarded and dysmorphic patient also had a small accessary ring derived from chromosome 4. FISH studies of this ring are consistent with an origin from a contiguous region including the centromere to band 4q12. It is apparent that there are at least two mechanisms for the formation of small ring chromosomes. This adds a further complication in any attempt to ascertain common phenotypes between patients known to have morphologically similar markers derived from the same chromosome. PMID- 7485166 TI - Sex chromosome loss and aging: in situ hybridization studies on human interphase nuclei. AB - A total of 1,000 lymphocyte interphase nuclei per proband from 90 females and 138 males age 1 wk to 93 years were analyzed by in situ hybridization for loss of the X and Y chromosomes, respectively. Both sex chromosomes showed an age-dependent loss. In males, Y hypoploidy was very low up to age 15 years (0.05%) but continuously increased to a frequency of 1.34% in men age 76-80 years. In females, the baseline level for X chromosome loss is much higher than that seen for the Y chromosome in males. Even prepubertal females show a rate of X chromosome loss, on the order of 1.5%-2.5%, rising to approximately 4.5%-5% in women older than 75 years. Dividing the female probands into three biological age groups on the basis of sex hormone function (< 13 years, 13-51 years, and > 51 years), a significant correlation of X chromosome loss versus age could clearly be demonstrated in women beyond age 51 years. Females age 51-91 years showed monosomy X at a rate from 3.2% to 5.1%. In contrast to sex chromosomal loss, the frequency of autosomal monosomies does not change during the course of aging: Chromosome 1 and chromosome 17 monosomic cells were found with a constant incidence of 1.2% and 1%, respectively. These data also indicate that autosome loss in interphase nuclei is not a function of chromosome size. PMID- 7485167 TI - Severe Gardner syndrome in families with mutations restricted to a specific region of the APC gene. AB - Familial adenomatous polyposis (FAP) is associated with a number of extraintestinal manifestations, which include osteomas, epidermoid cysts, and desmoid tumors, often referred to as "Gardner syndrome." Recent studies have suggested that some of the phenotypic features of FAP are dependent on the position of the mutation within the APC gene. In particular, the correlation between congenital hypertrophy of the retinal pigment epithelium (CHRPE) and APC genotype indicates that affected families may be divided into distinct groups. We have investigated the association between the dentoosseous features of GS on dental panoramic radiographs (DPRs) and APC genotype in a regional cohort of FAP families. DPRs were performed on 84 affected individuals from 36 families, and the dento-osseous features of FAP were quantified by a weighted scoring system. Significant DPR abnormalities were present in 69% of affected individuals. The APC gene mutation was identified in 27 of these families, and for statistical analysis these were subdivided into three groups. Group 1 comprised 18 affected individuals from seven families with mutations 5' of exon 9; these families (except one) did not express CHRPE. Groups 2 comprised 38 individuals from 16 families with mutations between exon 9 and codon 1444, all of whom expressed CHRPE. Group 3 comprised 11 individuals from four families with mutations 3' of codon 1444, none of whom expressed CHRPE. Families with mutations 3' of codon 1444 had significantly more lesions on DPRs (P < .001) and appeared to have a higher incidence of desmoid tumors. These results suggest that the severity of some of the features of Gardner syndrome may correlate with genotype in FAP. PMID- 7485168 TI - Human genetic susceptibility and infection with Leishmania peruviana. AB - Racial differences, familial clustering, and murine studies are suggestive of host genetic control of Leishmania infections. Complex segregation analysis has been carried out by use of the programs POINTER and COMDS and data from a total population survey, comprising 636 nuclear families, from an L. peruviana endemic area. The data support genetic components controlling susceptibility to clinical leishmaniasis, influencing severity of disease and resistance to disease among healthy individuals. A multifactorial model is favored over a sporadic model. Two locus models provided the best fit to the data, the optimal model being a recessive gene (frequency .57) plus a modifier locus. Individuals infected at an early age and with recurrent lesions are genetically more susceptible than those infected with a single episode of disease at a later age. Among people with no lesions, those with a positive skin-test response are genetically less susceptible than those with a negative response. The possibility of the involvement of more than one gene together with environmental effects has implications for the design of future linkage studies. PMID- 7485169 TI - Evidence for a major gene influencing 7-year increases in diastolic blood pressure with age. AB - The contribution of genetic factors to blood pressure levels is well established. The contribution of genes to the longitudinal change in blood pressure has been less well studied, because of the lack of longitudinal family data. The present study investigated a possible major-gene effect on the observed increase with age in diastolic blood pressure (DBP) levels. Subjects included 965 unmedicated adults (age > or = 18 years) in 73 pedigrees collected in Utah as part of a longitudinal cardiovascular family study. Segregation analysis of DBP change over 7.2 years of follow-up identified a recessive major-gene effect with a gene frequency of p = .23. There was also a significant age effect on the genotypic means, which decreased expression of the major gene at older ages. For those inferred to have the genotype responsible for large DBP increases, DBP increased 32.3%, compared with a 1.5% increase in the nonsusceptible group (P < .0001). The relative risk of developing hypertension between the susceptible and nonsusceptible groups after 7.2 years was 2.4 (P = .006). Baseline DBP reactivities to mental arithmetic (P < .0001), and isometric handgrip (P < .0001) stress tests were greatest in those assigned to the susceptible genotype. We conclude that age-related changes in DBP are influenced by a major gene. Characteristics of this major-gene effect for greater age-related blood pressure increases include greater reactivity to mental and physical stressors. The present study thus provides evidence for genetic control of changes in blood pressure, in addition to the previously suggested genetic control of absolute blood pressure level. PMID- 7485170 TI - Risk of chromosomal abnormalities, with emphasis on live-born offspring of young mothers. AB - In a large public urban hospital obstetrics service with > 123,000 deliveries in a 10-year period (1980-89), the frequencies (0.12%) of any type of chromosomal abnormality and of trisomy syndromes were analyzed for maternal age-related risk, by logistic regression. Focusing on very young gravidas, we found that in the study period there were 9,332 births (7.5% of all deliveries) to mothers < or = 16 years old. Estimated risks of chromosomal abnormalities among offspring associated with very young maternal age (9-16 years) were similar to those age associated risks of mothers 20-29 years old. Risks of chromosomal abnormalities increase with advancing maternal age and are independent of ethnicity. PMID- 7485171 TI - High diversity of alpha-globin haplotypes in a Senegalese population, including many previously unreported variants. AB - RFLP haplotypes at the alpha-globin gene complex have been examined in 190 individuals from the Niokolo Mandenka population of Senegal: haplotypes were assigned unambiguously for 210 chromosomes. The Mandenka share with other African populations a sample size-independent haplotype diversity that is much greater than that in any non-African population: the number of haplotypes observed in the Mandenka is typically twice that seen in the non-African populations sampled to date. Of these haplotypes, 17.3% had not been observed in any previous surveys, and a further 19.1% have previously been reported only in African populations. The haplotype distribution shows clear differences between African and non African peoples, but this is on the basis of population-specific haplotypes combined with haplotypes common to all. The relationship of the newly reported haplotypes to those previously recorded suggests that several mutation processes, particularly recombination as homologous exchange or gene conversion, have been involved in their production. A computer program based on the expectation maximization (EM) algorithm was used to obtain maximum-likelihood estimates of haplotype frequencies for the entire data set: good concordance between the unambiguous and EM-derived sets was seen for the overall haplotype frequencies. Some of the low-frequency haplotypes reported by the estimation algorithm differ greatly, in structure, from those haplotypes known to be present in human populations, and they may not represent haplotypes actually present in the sample. PMID- 7485172 TI - Toward fully automated genotyping: genotyping microsatellite markers by deconvolution. AB - Dense genetic linkage maps have been constructed for the human and mouse genomes, with average densities of 2.9 cM and 0.35 cM, respectively. These genetic maps are crucial for mapping both Mendelian and complex traits and are useful in clinical genetic diagnosis. Current maps are largely comprised of abundant, easily assayed, and highly polymorphic PCR-based microsatellite markers, primarily dinucleotide (CA)n repeats. One key limitation of these length polymorphisms is the PCR stutter (or slippage) artifact that introduces additional stutter bands. With two (or more) closely spaced alleles, the stutter bands overlap, and it is difficult to accurately determine the correct alleles; this stutter phenomenon has all but precluded full automation, since a human must visually inspect the allele data. We describe here novel deconvolution methods for accurate genotyping that mathematically remove PCR stutter artifact from microsatellite markers. These methods overcome the manual interpretation bottleneck and thereby enable full automation of genetic map construction and use. New functionalities, including the pooling of DNAs and the pooling of markers, are described that may greatly reduce the associated experimentation requirements. PMID- 7485173 TI - Comparative genomic hybridization in clinical cytogenetics. AB - We report the results of applying comparative genomic hybridization (CGH) in a cytogenetic service laboratory for (1) determination of the origin of extra and missing chromosomal material in intricate cases of unbalanced aberrations and (2) detection of common prenatal numerical chromosome aberrations. A total of 11 fetal samples were analyzed. Seven cases of complex unbalanced aberrations that could not be identified reliably by conventional cytogenetics were successfully resolved by CGH analysis. CGH results were validated by using FISH with chromosome-specific probes. Four cases representing common prenatal numerical aberrations (trisomy 21, 18, and 13 and monosomy X) were also successfully diagnosed by CGH. We conclude that CGH is a powerful adjunct to traditional cytogenetic techniques that makes it possible to solve clinical cases of intricate unbalanced aberrations in a single hybridization. CGH may also be a useful adjunct to screen for euchromatic involvement in marker chromosomes. Further technical development may render CGH applicable for routine aberration screening. PMID- 7485174 TI - Efficiency of typing unaffected relatives in an affected-sib-pair linkage study with single-locus and multiple tightly linked markers. AB - In an affected-sib-pair study, the parents are often unavailable for typing, particularly for diseases of late onset. In many cases, however, it is possible to sample unaffected siblings. It is therefore desirable to assess the contribution of such siblings to the power of such a study. The likelihood ratio introduced by Risch and improved by Holmans was extended to incorporate data from unaffected siblings. Tests based on two likelihoods were considered: the full likelihood of the data, based on the identity-by-descent (IBD) sharing states of the entire sibship, and a pseudolikelihood based on the IBD sharing states of the affected pair only, using the unaffected siblings to infer parental genotypes. The latter approach was found to be more powerful, except when penetrance was high. Typing an unaffected sibling, or just one parent, was found to give only a small increase in power except when the PIC of the marker was low. Even then, typing an unaffected relative increased the overall number of individuals that had to be typed to achieve a given power. If there is no highly informative marker locus in the area under study, it may be possible to "build" one by combining the alleles from two or more neighboring tightly linked loci into haplotypes. Typing two loci gave a sizeable power increase over a single locus, but typing further loci gave much smaller gains. Building haplotypes will introduce phase uncertainties, with the result that such a system will yield less power than will a single locus with the same number of alleles. This power loss was small, however, and did not affect the conclusions regarding the worth of typing unaffected relatives. PMID- 7485175 TI - Points to consider: ethical, legal, and psychosocial implications of genetic testing in children and adolescents. American Society of Human Genetics Board of Directors, American College of Medical Genetics Board of Directors. PMID- 7485176 TI - HLA and fertility. PMID- 7485177 TI - Our tangled heritage: demographic history and the Indian caste system. PMID- 7485180 TI - High-yield noninvasive human genomic DNA isolation method for genetic studies in geographically dispersed families and populations. PMID- 7485179 TI - Prevalence of Canavan disease heterozygotes in the New York metropolitan Ashkenazi Jewish population. PMID- 7485178 TI - Linkage disequilibrium at the Machado-Joseph disease/spinal cerebellar ataxia 3 locus: evidence for a common founder effect in French and Portuguese-Brazilian families as well as a second ancestral Portuguese-Azorean mutation. PMID- 7485181 TI - Contamination of cDNA- libraries and expressed-sequence-tags databases. PMID- 7485182 TI - Internet solicitation of research subjects for genetic studies. PMID- 7485183 TI - The high prevalence of fragile X premutation carrier females: is this frequency unique to the French Canadian population? PMID- 7485184 TI - Low-sister-chromatid-exchange Bloom syndrome cell lines: an important new tool for mapping the basic genetic defect in Bloom syndrome and for unraveling the biology of human tumor development. PMID- 7485186 TI - Cancer risk due to occupational exposure to polycyclic aromatic hydrocarbons. AB - Polycyclic aromatic hydrocarbons (PAHs) demonstrate carcinogenic activity in animal models. Although some epidemiologic studies have implicated PAHs as risk factors for human cancer, the evidence reported to date has not been consistent. The purpose of this report is to describe the associations between occupational exposure to PAHs in the workplace and each of 14 types of cancer. A population based, case-control study was carried out in Montreal to investigate associations between a large variety of environmental and occupational exposures on the one hand, and several types of cancer on the other. A detailed job history was obtained from each subject along with information on a number of potential confounders. Each job history was reviewed by a team of experts, who used this information to construct a corresponding history of occupational exposures. Among the PAH exposures considered were benzo(a)pyrene (B(a)P) and five categories of PAHs defined on the basis of the source material, namely, wood, petroleum, coal, other sources, and any source. Altogether, 3,730 cancer patients and 533 population controls were interviewed and their job exposure histories coded. For each of 14 types of cancer analyzed, three control groups were available: other cancer patients, population controls, and the pooled set of cancer and population controls. The associations between 14 cancer types and 6 PAH exposures were analyzed using logistic regression methods. For most types of cancer evaluated, there was no evidence of excess risk due to PAHs at the levels encountered in the occupations in which PAH exposure has been prevalent in the Montreal area. For a few cancer sites--the esophagus, the pancreas, and the prostate gland--there were suggestions of excess risk; these observations are noteworthy hypotheses for further investigation. For lung cancer, there appeared to be an increased risk due to PAHs among nonsmokers and light smokers, but not among heavy smokers. PMID- 7485187 TI - Mortality patterns among women in the motor vehicle manufacturing industry. AB - This retrospective follow-up study evaluated the mortality experience of 11,271 women employed in a large motor vehicle manufacturing company in 1973. Company computerized personnel records were used to identify subjects and to obtain their employment histories. Follow-up through December 31, 1985, was conducted using company, state, and national data sources. The mortality rates of the cohort were compared with the rates of the U.S. general population of white or black women, adjusting for age and calendar time and using the standardized mortality ratio (SMR) as the measure of association. The overall mortality rate of the cohort was lower than the U.S. rate by 15% among whites (SMR = 85, 95% confidence interval (CI) = 78-93) and by 40% among blacks (SMR = 59, CI = 49-70). Among whites, the low mortality rate was due primarily to deficits of noncancer deaths, although a 32% deficit also was observed for breast cancer (SMR = 68, CI = 48-92). Black female workers had 40% fewer deaths than expected overall, and their all cancer SMR also was below expectation (SMR = 82, CI = 60-109). Despite evidence of a strong healthy worker effect among women in the motor vehicle manufacturing industry, the study found excesses of lung cancer among white women in assembly jobs (SMR = 158, CI = 107-226) and of pancreatic cancer among white assembly workers in upholstery and trim operations (SMR = 302, CI = 97-704). PMID- 7485185 TI - New genes for boys. PMID- 7485189 TI - Occupational risk factors for lung cancer in Tianjin, China. AB - Information about occupation and industry of cancer patients is routinely recorded in the Cancer Registry of Tianjin, People's Republic of China. Occupational risk factors of lung cancer for the period of 1981-1987 were assessed using other cancers as controls. For each occupation and industry, workers employed in it were considered exposed and were compared with all other workers. Elevated risks were found for a number of industries for both sexes, including, in particular, the textile, wood, metal, and construction industries. Results by occupation were similar to those by industry. Smoking scores were calculated for various occupations on the basis of a separate population survey in Tianjin. Smoking cannot explain the elevated odds ratios in males, while among females a strong correlation was observed between smoking scores and lung cancer odds ratios, suggesting a confounding effect. PMID- 7485188 TI - Occupational respiratory cancer and exposure to asbestos: a case-control study in a cohort of workers in the electricity and gas industry. AB - Cancers of the pleura, lung, and larynx between 1978 and 1989 among active male workers of Electricite de France-Gaz de France were studied in association with asbestos exposure using a case-control design nested within the cohort of workers of the company. The cohort included about 1,400,000 person-years, corresponding to a mean of 117,000 men per year. Exposure to asbestos and to some potential occupational confounders selected among agents from groups I, IIa, and IIb of the International Agency for Research on Cancer was assessed by a job-exposure matrix specific to the company. During the observation period, 12 cases of pleural cancer, 310 cases of lung cancer, and 116 cases of larynx cancer were registered in the cancer register of the company social security department. Four controls per case, matched for year of birth, were randomly selected among the cohort. Conditional logistic regression was used to estimate the odds ratios. A first analysis was conducted in order to assess the validity of the job-exposure matrix by investigating already known relationships between asbestos exposure and asbestosis. For asbestosis, a strong exposure-response relation was found with an odds ratio (OR) of 57.4 [95% confidence interval (CI): 17.0-194.0] in the highest exposure group. There was an elevated risk of pleural cancer (OR, 4.8, CI, 1.2 19.8). For lung cancer, significant ORs of 2.0 (CI, 1.3-3.2) and 1.9 (CI, 1.2 3.0) were found among the two highest cumulative exposure groups; adjustement for confounders slightly decreased the ORs. Squamous cell neoplasm of the lung was associated with asbestos exposure. The association between larynx cancer and asbestos exposure showed a tendency towards a nonsignificant increase in ORs in the highest cumulative exposure categories; this tendency disappeared when adjusting for occupational confounders. This study showed that occupational exposure to asbestos could increase the risk of pleural and lung cancer in a sector in which exposure levels are not considered to be high compared with other industrial settings. PMID- 7485191 TI - Mortality among workers at a plastics manufacturing and research and development facility: 1946-1988. AB - Mortality through 1988 was studied for 5,932 male employees who worked between January 1, 1946 and December 31, 1967 at a New Jersey plastics manufacturing and research and development facility. The cohort was followed for an average of 32 years and included 1,859 deaths. Potential exposures included asbestos, formaldehyde, and polyvinyl chloride (PVC). Mortality rates for the cohort were compared to both U.S. and state mortality rates, and analyses were also performed by lagging duration of employment. Based on U.S. rates, mortality among hourly males (n = 3,853) from all cancers was similar to expected [standardized mortality ratio (SMR), 102; 95% confidence interval (CI), 92-114]. Excess mortality among hourly workers was seen for pancreatic cancer (SMR, 146; 95% CI, 95-216) and "malignancies of other parts of the respiratory system" (SMR, 373; 95% CI, 121-870). The latter excess was due entirely to five deaths from pleural mesothelioma. There were no deaths identified due to nasal cavity or nasopharyngeal cancers, or angiosarcoma of the liver. Mortality from leukemia among research and development workers (n = 1,421) was significantly elevated (SMR, 265; 95% CI, 115-524) and related to assignment to process development. This study verifies the excess of pancreatic cancer among workers at the facility seen in earlier studies and observes excesses of mesothelioma due to asbestos exposure and leukemia in process development workers. PMID- 7485190 TI - Asbestos-induced visceral pleural fibrosis reduces pulmonary compliance. AB - To determine the nature of respiratory functional impairment caused by asbestos induced visceral pleural fibrosis (VPF) and to discover which pulmonary physiological variable best reveals it, we examined 59 asbestos-exposed construction workers having asbestos-related changes on chest radiographs. Computed tomography scans of the thorax were also performed. Visceral pleural fibrosis was diagnosed in 29 subjects: seven had only VPF, 17 had VPF and pleural plaques, and five had VPF, plaques, and asbestosis. In subjects without VPF, 23 had plaques, six had plaques and asbestosis, and one had only minor fibrotic parenchymal changes insufficient for a diagnosis of asbestosis. Flow-volume spirometry, body plethysmography, static and dynamic compliance, and pulmonary diffusing capacity for carbon monoxide were measured. The subjects with VPF had significantly lower static (p = 0.005) and dynamic (p = 0.007) compliance values than those without. Other respiratory function variables failed to show any significant differences. We conclude that the measurement of static and dynamic compliance is a useful method in assessing pulmonary function impairment caused by visceral pleural fibrosis. PMID- 7485192 TI - Pancreatic cancer among workers processing synthetic resins. AB - A nested case-control study was conducted to investigate whether an excess of pancreatic cancer, identified in a cohort mortality study with follow-up from 1946 through 1988, was associated with potential workplace exposures at a New Jersey plastics manufacturing and research and development facility. The study population included 28 male pancreatic cancer cases and 140 randomly selected controls, matched on year of birth and at risk (alive) at the time of the case death. Using plant work history records, department assignments for the two groups were compared according to duration and time since first assignment. Workers assigned to a work area that processed vinyl resins and polyethylene (PE) were shown to be at increased risk. Men assigned more than 16 years to this department had a significantly increased risk ratio of 7.15 (95% confidence intervals [CI]: 1.28-40.1). No excess was seen with shorter duration assignments. Seven of the nine cases began working in this area in the 1940s. Average latency was 32 years, and all but three cases worked 20 years or more in this unit. Over the study period, significant exposure-related process changes occurred, in addition to the use of numerous chemical additives. Although vinyl and PE processing operations could not be analyzed separately, the pancreatic cancer excess is more likely to be related to vinyl processing. Identification of a causative agent or combination of agents would require investigations with more detailed exposure information. PMID- 7485193 TI - Update of the mortality experience of employees with occupational exposure to 1,2 dibromo-3-chloropropane (DBCP). AB - DBCP (1,2-dibromo-3-chloropropane), a nematocide, was used in the United States from the mid-1950s until 1977. The U.S. Environmental Protection Agency (EPA) restricted and eventually banned its use after the 1977 discovery of DBCP-induced sterility in production workers. The present study is an update of the mortality (1957-1989) experience of a cohort of 548 male employees who had potential for exposure in the production and formulation of DBCP. While adjusting for age, calendar-year, and pay status of all other Midland-area Dow Chemical male employees, there were 68 total observed deaths in the cohort compared to 72.1 expected (Mantel Haenszel Relative Risk 0.9, 95% Confidence Interval 0.7-1.2) and 19 deaths from all malignancies compared to 19.0 expected (RR 1.0, 95% CI 0.6 1.6). Of the a priori anatomic cancer sites of interest, there were no deaths from stomach, liver, kidney, testes, or nasal cavity cancers. Altogether, there were 7 deaths from lung cancer compared to 6.6 expected (RR 1.1, 95% CI 0.5-2.3). Among the 81 employees with exposure categorized as direct for 1 or more years, there were 3 observed lung cancer deaths compared to 0.9 expected (RR 3.3, 95% CI 1.1-9.6). Smoking was a confounding factor in the interpretation of this observation. Although the present analysis nearly doubled the number of person years from the original study, the conclusions remain limited by the cohort's size and duration of follow-up. PMID- 7485194 TI - Prevalence and predictors of asthma in working groups in British Columbia. AB - We evaluated the prevalence of asthma and its predictors in studies of several male working groups: 619 cedar sawmill, 724 grain elevator, 399 pulpmill, 798 aluminum smelter, and 1,127 unexposed workers. These workers had taken part in health studies for assessment of chronic respiratory effects of various workplace exposures between 1979 and 1982. The American Thoracic Society Adult Questionnaire (ATS-DLD-78) was used for these studies. Allergy skin tests were also performed. The participation rates were > 80%. The overall prevalance of physician-diagnosed asthma was 4.6%, and current asthma 3%. The prevalence of asthma after employment in the current industry, as a surrogate for work-related asthma, was 3.9 times higher in cedar sawmill workers, 2.2 times higher in pulpmill and aluminum smelter workers, and 1.7 times higher in grain elevator workers compared with unexposed workers. Atopy and a positive parental history of asthma, but not smoking, were important risk factors for asthma before the onset of first employment. Also, for asthma after employment in the current industry, atopy and a positive parental history of asthma were important risk factors. Smoking was associated with a significant reduction in the risk for asthma after employment in the current industry. Within specific work groups, the prevalence of atopy was significantly higher among pulpmill workers with asthma after employment in current industry than those without asthma. Conversely, cedar sawmill workers who had asthma after employment in the current industry were nonatopic and nonsmokers. PMID- 7485195 TI - Three cases of idiopathic interstitial pneumonia with bullae seen in schoolteachers. AB - We encountered three patients with chronic interstitial pneumonia with many bullae in the lower lung fields whose lifetime occupation was teaching school. Pathological examination of autopsy lungs of these patients revealed interstitial pneumonia and multiple bullae throughout the lungs, including the lower lobe. Since blackboard chalk has been used as a popular writing material among teachers in Japan, the mineral contents in the lungs of two of the three cases and four control cases with idiopathic interstitial pneumonia (IIP) (whose occupations were not teaching) were analyzed. The amount of deposition of total dust, inorganic dust, non-SiO2 inorganic dust, and calcium was significantly higher in the lungs of two schoolteachers compared with those of the control lungs. The amount of free silica in case 1 and alpha-quartz in case 3 were also significantly higher than in the controls. Two thirds of the chalk produced in Japan is still made from gypsum and involves small amounts of silica and other minerals, in addition to calcium. These findings indicated the deposition of chalk in the lungs of these patients with interstitial pneumonia and multiple bullae. PMID- 7485196 TI - Bronchiolitis obliterans in an animal feed worker. AB - A nonsmoking animal feed worker developed severe irreversible airflow obstruction during a 2-year occupational exposure to organic matter, microorganisms, proteolytic enzymes, and both amorphous and synthetic silicates. Her pulmonary dysfunction failed to improve despite removal from the workplace and treatment with bronchodilators and corticosteroids. Open lung biopsy demonstrated peribronchiolar inflammation, scarring within the small airways, and neolumen formation, findings consistent with bronchiolitis obliterans. Energy dispersive analysis of both the bronchoalveolar lavage fluid as well as the biopsy specimens revealed the presence of silicate particles, suggesting a potential link between the silica exposure and the pathologic findings. This case suggests bronchiolitis obliterans can occur as a consequence of occupational exposure in the animal feed industry. In addition, this entity should henceforth be considered when evaluating symptomatic patients with exposure to amorphous and synthetic silicates. PMID- 7485198 TI - Interdisciplinary clerkships: educational models of the future? PMID- 7485197 TI - Immunological reactions and respiratory function in wool textile workers. AB - Immunological status and respiratory function were studied in a group of 64 wool textile workers (52 women and 12 men). A group of 46 workers not exposed to wool dust served as a control for the respiratory symptoms and immunologic testing. Skin testing was performed with different wool allergens (domestic and Australian) as well as with common allergens. Ventilatory capacity was measured in wool workers on Mondays before and after the work shift. The prevalence of positive skin tests to all allergens was higher in wool than in control workers, although the difference was statistically significant only for washed domestic wool (wool workers: 42.2%; control workers: 19.6%; p < 0.05). Increased serum IgE levels were more frequent in wool (26.6%) than in control workers (3.1%) (p < 0.01). In wool textile workers there was a high prevalence of acute and chronic respiratory symptoms. Significant across-shift reductions in ventilatory capacity tests, as well as abnormal baseline lung function, were recorded in wool textile workers. Individual data demonstrated that many of the wool workers had FEF25 lower than 70% of predicted. In general, the prevalence of symptoms and the lung function abnormalities did not correlate with the results of specific (wool) skin tests. Our data indicate that exposure to wool dust in some workers may be associated with the development of acute and chronic respiratory symptoms and impairment of lung function. Immunologic abnormalities, although frequent in this group, do not appear to be associated with the severity of these changes. PMID- 7485199 TI - Measuring--and then strengthening--the public pulse about medical research. PMID- 7485200 TI - Body fat distribution and dyslipidemia. PMID- 7485201 TI - Body fat distribution, rather than overall adiposity, influences serum lipids and lipoproteins in healthy men independently of age. AB - PURPOSE: We investigated the relationships between the amount and distribution of body fat and fasting serum lipids and lipoproteins to explore whether coronary artery disease (CAD) risk may be mediated through effects on the serum lipid profile. PATIENTS AND METHODS: We determined serum total cholesterol and triglyceride, low-density lipoprotein cholesterol, high-density lipoprotein (HDL) cholesterol, and HDL subfractions 2 and 3 in 103 healthy men, aged 21 to 77 years (mean 48.7). The amount and distribution of fat were determined directly by dual energy X-ray absorptiometry. Adiposity was determined as the ratio between total body fat tissue and total body lean tissue, while fat distribution was taken as the ratio between the mass of fat tissue in the android (central) and gynoid (hip and thigh) regions. RESULTS: Univariate analysis showed both adiposity and fat distribution to be correlated with total serum cholesterol and triglyceride concentrations (adiposity r = .20, .21; both P < 0.05: fat distribution r = .25, .38; P < 0.05, P < 0.001, respectively). Fat distribution was also negatively correlated with HDL2 cholesterol (r = -.20, P < 0.05). In multiple linear regression analysis, neither age nor adiposity was significantly correlated with any serum lipid or lipoprotein concentration, while increasing android-to-gynoid ratio was independently associated with elevated total serum triglyceride (r = .40, P < 0.01) and decreased HDL2 (r = -.25, P < 0.05) concentrations. CONCLUSIONS: The association of both age and overall adiposity with the fasting serum lipid profile are mediated via their correlations with body fat distribution. In men, the distribution, rather than the amount, of body fat is related to adverse changes in serum lipids and lipoproteins, and hence potentially to increased CAD risk. PMID- 7485202 TI - Lupus hepatitis: an under-recognized disease feature associated with autoantibodies to ribosomal P. AB - PURPOSE: To determine the frequency and clinical manifestations of systemic lupus erythematosus (SLE)-related liver disease and to establish whether this disease feature correlates with circulating autoantibodies to ribosomal P proteins. PATIENTS AND METHODS: A retrospective chart review of a large lupus cohort searching for laboratory and clinical manifestations of liver disease. A case control study with testing of stored serum for antiribosomal P antibodies using immunoblotting, an enzyme-linked immunosorbent assay, and immunodiffusion in cases of lupus with liver involvement, and in randomly selected and ethnically matched controls with lupus but without liver involvement. RESULTS: Of 131 patients with SLE, 4 (3%) had liver involvement that could only be ascribed to the disease itself, and 2 additional cases from elsewhere were also studied. The clinical picture of the liver involvement resembled chronic active hepatitis, but there was no serological evidence of hepatitis B or C infections. Only 1 patient had low-titer antismooth muscle antibodies, and none had antimitochondrial antibodies. Antiribosomal P antibodies were present in all 6 patients with lupus hepatitis, compared to only 2 (10%) of 20 of controls with lupus but no liver disease (P = 0.0001, odds ratio 96). CONCLUSIONS: Lupus hepatitis appears to be an infrequent but distinct manifestation of SLE which correlates strongly with the presence of antiribosomal P antibodies. Its course and prognosis are variable, ranging from chronic biochemical abnormalities of liver function to acute clinical hepatitis to hepatic failure. PMID- 7485203 TI - A family study of anticardiolipin antibodies and associated clinical conditions. AB - PURPOSE: To determine the frequency of anticardiolipin antibodies (aCL) and their clinical sequelae in family members of aCL-positive patients. PATIENTS AND METHODS: A prospective serologic and clinical evaluation was performed on 23 patients with elevated aCL titers, 87 blood relatives, 18 spouses, and 37 controls. aCL and antinuclear antibodies (ANA) were measured and clinical histories were assessed for all probands, relatives, spouses, and controls. RESULTS: Fifty of 87 relatives screened (57%) had evidence of autoantibody production (aCL and/or ANA). Twenty-nine (33%) had positive aCL titers. Twenty were positive for aCL-immunoglobulin (Ig) G, 7 had evidence of both aCL-IgG and aCL-IgM, and an additional 2 were positive for aCL-IgM alone. In contrast, only 1 spouse was aCL-IgG positive. Thirty-two relatives and 1 spouse were ANA positive. All controls were negative for aCL and ANA. Significant differences were noted between relatives and spouses for aCL-IgG (P < 0.00001) and aCL-IgM titers (P < 0.0066), and also between relatives and controls (P < 0.00001 for both). Clinically, 4 cases of systemic lupus erythematosus (SLE), 4 SLE-like diseases, and 8 aCL-associated illnesses (2 premature strokes, 3 recurrent fetal losses, 1 recurrent thrombosis, and 2 cases of thrombocytopenia) were documented in the relatives. All cases were associated with aCL and/or ANA production. CONCLUSIONS: ANA, aCL, and clinical events associated with antiphospholipid antibodies occur with increased frequency in relatives, but not spouses of aCL-positive probands. These results suggest that aCL-related illnesses may be familial. PMID- 7485204 TI - Autoimmune thyroid disease in primary Sjogren's syndrome. AB - PURPOSE: To evaluate the prevalence of autoimmune thyroid disease and thyroid dysfunction in patients with primary Sjogren's syndrome. PATIENTS AND METHODS: Thyroid function of 33 patients with primary Sjogren's syndrome was clinically and biochemically evaluated. Thyroid hormones and autoantibodies against thyroid peroxidase, thyroglobulin, and thyroid hormones were measured. RESULTS: Autoimmune thyroid disease and thyroid dysfunction were found in 15 cases (45%): autoimmune thyroiditis in 8 (24%); autoimmune hyperthyroidism in 2 (6%); and reversible iodine-induced hypothyroidism in the remaining 5 (15%). One or more of the evaluated autoantibodies were detected in 8 euthyroid patients (24%). Overall, the prevalence of autoantibodies against thyroid peroxidase, thyroglobulin, thyroxine, and triiodothyronine was 45%, 18%, 42%, and 36%, respectively. CONCLUSIONS: The high prevalence of autoimmune thyroid disease and thyroid dysfunction found in primary Sjogren's syndrome, using sensitive immunologic and thyroid function tests, suggest that both diseases are more frequently associated than it was previously thought, and should be sought clinically and by laboratory tests in all patients with primary Sjogren's syndrome. PMID- 7485205 TI - Lipid metabolism and apolipoprotein E phenotypes in patients with xanthelasma. AB - PURPOSE: To know the prevalence and types of dyslipidemia associated with xanthelasma. PATIENTS AND METHODS: One hundred fifteen patients with xanthelasma and 105 age-matched control subjects without xanthelasma were evaluated in a cross-sectional study. Univariate and multivariate comparisons of lipid variables (including total cholesterol; triglycerides; very-low-, low-, and high-density lipoprotein cholesterol [VLDL-C, LDL-C, and HDL-C, respectively]; cholesterol of high density lipoprotein [HDL] subfractions 2 and 3 [HDL2-C and HDL3-C]; apolipoprotein (apo) A-I and B; and apo E phenotypes) and nonlipid coronary risk factors were made between patients with and without xanthelasma. RESULTS: Patients with xanthelasma had higher levels of cholesterol, LDL-C, and apo B, and lower levels of HDL2-C than control subjects. The prevalence of the apo E4/E3 phenotype was higher in cases than in controls (P < 0.05). Patients with xanthelasma had a higher prevalence of personal and familiar history of cardiovascular disease and were more overweight than control subjects. A stepwise discriminant analysis disclosed an independent association of xanthelasma with lower HDL-C, HDL2-C, and HDL3-C levels in men, and with higher total cholesterol and lower HDL2-C levels in women. CONCLUSIONS: Xanthelasma appears to be associated with qualitative and quantitative abnormalities of lipid metabolism that may favor lipid deposition in the skin and arterial wall. The findings support the notion that xanthelasma is a marker of dyslipidemia, and underline the need to determine a full lipid profile in these patients to detect those potentially at increased risk of cardiovascular disease. PMID- 7485206 TI - The solitary pulmonary nodule: update 1995. AB - PURPOSE: This study analyzed the clinical characteristics, diagnostic evaluation, prevalence of malignancy, and outcome of patients with a solitary pulmonary nodule (SPN) encountered in the outpatient practice of a pulmonologist in an urban university hospital from 1990 to 1993. PATIENTS AND METHODS: SPN was defined as a round or ovoid density < or = 3 cm in diameter within the lung parenchyma. Patients with and without lung cancer in SPNs were compared. RESULTS: Forty patients had a mean age of 65 years, an almost equal sex distribution, high prevalences of cardiovascular disease (53%) and chronic obstructive pulmonary disease (COPD) (33%), but a low incidence of tuberculosis. The mean size of SPNs was 1.8 cm. The prevalence of malignancy was 53%. In SPNs < or = 2 cm in diameter, the prevalence of malignancy was 43%. Nonsurgical biopsy techniques made a diagnosis in 78% of patients. In 94% of patients with lung cancer in SPNs, the tumor was resectable (stage 1, 2, or 3A), emphasizing the need for early detection. Despite the small size of the SPNs, the prevalence of malignancy was high. CONCLUSION: Despite the advanced age and high prevalence of cardiovascular disease and COPD in patients with SPNs, lung cancer that occurs in these lesions appears to have a favorable prognosis if detected promptly. PMID- 7485207 TI - The beneficial effect of angiotensin-converting enzyme inhibition with captopril on diabetic nephropathy in normotensive IDDM patients with microalbuminuria. North American Microalbuminuria Study Group. AB - PURPOSE: To determine whether angiotensin-converting enzyme (ACE) inhibition with captopril reduces the progression of microalbuminuria to overt proteinuria in normotensive patients with insulin-dependent diabetes mellitus (IDDM). PATIENTS AND METHODS: This study was a prospective randomized, double-blind, placebo controlled trial involving 26 centers in the United States and Canada. One hundred forty-three subjects, 14 to 57 years of age, with IDDM for 4 to 33 years, blood pressure < 140/90 mm Hg in the absence of antihypertensive therapy, and persistent albumin excretion 20 to 200 micrograms/min were randomized to double blind treatment with captopril 50 mg or placebo BID. Albumin excretion rate (AER), blood pressure, and glycohemoglobin were determined every 3 months, and creatinine clearance (CrCl) and urea excretion were measured every 6 months. RESULTS: Within 24 months, 6.0% (4/67) of captopril-treated subjects and 18.6% (13/70) of placebo-treated subjects progressed to clinical proteinuria, defined as AER > 200 micrograms/min and at least 30% above baseline (risk reduction = 67.8%, P = 0.037). AER increased at an annual rate of 11.8% (95% confidence interval [CI] -3.3% to 29.1%) in the placebo group, while it declined by 17.9% (CI -29.6% to -4.3%) in the captopril group (P = 0.004). CrCl decreased by 4.9 mL/min per 1.73 m2 per year in the placebo group, while it remained stable in the captopril group (0.9 mL/min per 1.73 m2 per year, P = 0.039 between groups). Ten subjects required treatment for hypertension; 8 in the placebo group and 2 in the captopril group. There was little correlation between the 24-month changes in mean arterial blood pressure and AER in either group. Glycohemoglobin and urinary urea excretion did not differ between groups. CONCLUSIONS: After 24 months of therapy with captopril, compared with placebo, normotensive subjects with IDDM experienced significantly less progression of microalbuminuria to clinical proteinuria, reduced albumin excretion, and preserved CrCl rate. The ACE inhibitor, captopril, was well tolerated. PMID- 7485208 TI - Psychiatric illnesses in patients with syncope. AB - PURPOSE: To determine the prevalence of psychiatric disorders among patients with syncope; the relationship between psychiatric disorders and the cause of syncope; and the outcome of syncope patients with psychiatric disorders. PATIENTS AND METHODS: In a prospective cohort study in a university-affiliated medical center, hospitalized and ambulatory patients with syncope underwent a standardized medical and psychiatric evaluation consisting of the Diagnostic Interview Schedule (DIS) sections for somatization, panic, generalized anxiety, major depression disorders, and drug and alcohol dependence and abuse disorders. RESULTS: Of 414 patients who completed the DIS, 82 (20%) met the criteria for at least one of the psychiatric disorders or alcohol/drug disorders. Patients with at least one of the four major psychiatric disorders (n = 58) were more likely to report > or = 4 syncopal events in the past year (odds ratio (OR) 2.5, P < 0.04); and to have prodromal symptoms prior to syncopal events (OR 2.4, P < 0.04) than were patients without psychiatric disorders. Syncope patients with only alcohol/drug disorders (n = 24) were more likely to be male (OR 7.7, P < 0.001) and younger than 65 years old (OR 4.5, P < 0.001) than patients without alcohol/drug disorders. One-year recurrence rate for syncope in patients with any psychiatric diagnosis was 35%, compared to 15% in those without a psychiatric disorder (P < 0.0001). Physicians were able to recognize potential psychiatric or alcohol/drug disorders in 70% of patients with more than one psychiatric disorder, but failed to note or treat 60% of the patients with only one of the assessed disorders. CONCLUSIONS: We recommend screening for psychiatric disorders in patients with unexplained syncope, especially in individuals with recurrent syncope and multiple physical symptoms, or for males who are under 65 years of age. PMID- 7485209 TI - Correlates of regular laxative use by frail elderly persons. AB - PURPOSE: To examine the demographic, clinical, and pharmacological correlates of regular laxative use in elderly persons residing in a long-term care setting. METHODS: This was a cross-sectional study using retrospective record review undertaken in an academically affiliated long-term care facility in the United States. All individuals residing in the institution for at least 1 month (n = 694) were characterized regarding use of laxatives. Regular laxative use was defined as more than 30 doses of laxatives, stool softeners, or enemas taken over the most recent 1-month period. RESULTS: Residents with regular laxative use (n = 349) were compared with those who received no laxatives (n = 227). Factors significantly associated with regular laxative use at the P < 0.05 significance level were simultaneously included in a multiple logistic regression model. Factors associated with regular laxative use were immobility, Parkinson's disease, diabetes mellitus, and use of iron supplements, calcium channel blockers, and antidepressants with moderate to strong anticholinergic properties. CONCLUSION: Regular laxative use is often associated with neurologic dysfunction that directly or indirectly affects the gut, or medications known to depress colonic motility. Identification of potentially modifiable correlates of regular laxative use in older individuals may suggest management strategies to avoid or reduce laxative, stool softener, and enema requirements, improve constipation symptoms, and enhance quality of life for the frail elderly population. PMID- 7485210 TI - Screening family members of patients with hereditary hemorrhagic telangiectasia. AB - PURPOSE: Hereditary hemorrhagic telangiectasia (HHT) is an autosomal dominant inherited disorder which may give rise to arteriovenous malformations in lungs and brain. When left untreated, these may cause serious complications. We screened family members of HHT patients for presence of the disease and associated pulmonary or cerebral arteriovenous malformations. PATIENTS AND METHODS: We investigated 98 family members of HHT patients on an outpatient basis. A stepped screening protocol was used based on prevalence of different manifestations of HHT. RESULTS: Thirty-six cases of HHT were found in the 98 persons screened. Pulmonary arteriovenous malformations were found in 12 of the 36 patients (33%), and cerebral arteriovenous malformations in 4 (11%). Therapy was recommended in 9 patients with pulmonary arteriovenous malformations and in 2 with cerebral arteriovenous malformations. CONCLUSIONS: Family members of known HHT patients should be encouraged to engage in a screening program, since the prevalence of potentially serious localizations is higher than previously thought. PMID- 7485212 TI - Experience with community academic practice: strategies for the ambulatory education of residents. AB - The expectation that training programs will place more of their physician graduates into primary care has caused internal medicine programs to explore different educational experiences for their residents. One proposal is to transfer the ambulatory training site for residents from the hospital clinic to a community academic practice (CAP) site. We discuss the theoretical aspects, the practical considerations, and our experience with one CAP site, including the problems that have been solved and the problems remaining. PMID- 7485211 TI - Treatment of orthostatic tachycardia with erythropoietin. AB - PURPOSE: To determine whether increasing red blood cell volume with erythropoietin reverses the hemodynamic response to standing in patients with orthostatic tachycardia. PATIENTS AND METHODS: Eight patients (2 men, 6 women) with orthostatic tachycardia were administered erythropoietin (50 U/kg body weight 3 times a week for 6 to 12 weeks) in order to reverse their red blood cell volume deficit. Six of the patients also received fludrocortisone (0.1 mg/d). Plasma and red blood cell volumes as well as the hemodynamic response to orthostatic stress were measured before and after erythropoietin therapy. RESULTS: Erythropoietin therapy increased the mean +/- hematocrit from 37.6 +/- 1.0 to 46.4 +/- 1.4 (+/- standard error) (P < 0.01) and increased the red blood cell volume from 17.7 +/- 1.1 to 24.6 +/- 2.0 mL/kg (P < 0.01). Treatment increased supine mean blood pressure (from 87 +/- 4 to 93 +/- 5 mm Hg, P < 0.025) and standing mean blood pressure (from 87 +/- 4 to 94 +/- 5 mm Hg, P < 0.025). Erythropoietin therapy, however, failed to reverse orthostatic tachycardia. Following treatment, the mean heart rate after 5 minutes standing was 129 +/- 7 bpm, not significantly different from the pretreatment standing heart rate (134 +/- 5 bpm). CONCLUSIONS: Although patients with the orthostatic tachycardia syndrome have a deficit in red blood cell volume, this is not the cause of their abnormal hemodynamic response to standing. Erythropoietin therapy fails to reverse orthostatic tachycardia. PMID- 7485213 TI - Recent advances in the application of gene therapy to human disease. AB - PURPOSE: To review the recent advances in the application of genetic modification strategies to the therapy of human diseases for which a molecular defect is known. METHODS: A computerized data bank search, the minutes of the National Institutes of Health (NIH) Recombinant DNA Advisory Committee published in the Federal Record, and reports of human clinical trials were used as data sources for this review. Clinical trials included in this review were published in the literature or approved by the NIH Recombinant DNA Advisory Committee. STUDY SELECTION: Evaluations of the efficacy of genetic modification strategies in clinical trials in human and in animal models are summarized. The design and outcome of the genetic modification strategies employed are reviewed for 16 marking trials, 16 gene replacement trials for molecular deficiency diseases, 3 chemoprotection and 4 chemotherapy sensitization trials, 11 cancer vaccine trials, 2 antisense oligonucleotide trials, and 3 molecular immunotherapy trials. DATA SYNTHESIS: The marking trials have shown that residual leukemia cells in the infused autologous marrow can contribute to relapse following autologous bone marrow transplants. The use of genetic modification for the replacement of missing or deficient genes in severe combined immunodeficiency, familial hypercholesterolemia, and cystic fibrosis has been associated with encouraging results so far. Clinical genetic therapy trials involving cancer vaccines, antisense oligonucleotides, adoptive immunotherapy with genetically modified T cells, delivery vectors containing interleukin-1 receptor inhibitor for arthritis, replacement strategies for storage diseases, and genetic suppression of human immunodeficiency viral replication are just commencing. CONCLUSIONS: The clinical application of genetic modification techniques has thus far been successful in the beginning phases of this field. These early results suggest that continuation of gene therapy trials designed to correct the molecular changes that lead to disease states in humans is warranted. Evaluation of such clinical trials in the future may be based on the analysis of assays for short term surrogate endpoints, as well as on the therapeutic outcomes of the trial, such as survival or remission. PMID- 7485214 TI - Mechanical ventilation: a review for the internist. PMID- 7485215 TI - Nephrotic syndrome, nephromegaly, renal hyperfiltration, and sudden death in a diabetic woman. PMID- 7485216 TI - Ventricular tachycardia as a cause of syncope in a young woman with a positive tilt-table test and a negative electrophysiologic study. PMID- 7485217 TI - Autoimmune thyrotoxicosis during lithium therapy in a patient with manic depressive illness. PMID- 7485218 TI - Conceptual and cost analysis errors in the evaluation of bedside capillary glucose testing. PMID- 7485219 TI - Mortality of untreated primary hyperparathyroidism--a nontraditional indication for parathyroid surgery? PMID- 7485220 TI - High retinol/retinol-binding protein ratio in noninsulin-dependent diabetes mellitus. AB - The authors evaluated serum retinol, retinol-binding protein (RBP), and beta carotene levels to elucidate the retinoid metabolism in non-insulin-dependent diabetes mellitus (NIDDM). The mean retinol levels by gender (1.83 mumol/L for females and 2.24 mumol/L for males) in diabetics were higher than those (1.31 mumol/L for females and 1.82 mumol/L for males) in control subjects (P < 0.0001, P < 0.01, respectively). The mean retinol/RBP ratios (0.95 for females and 0.97 for males) of diabetics were higher than those of the control subjects (0.60 for females and 0.64 for males) and of male patients having impaired glucose tolerance (0.55) (P < 0.0001). Lipid-lowering medication significantly decreased retinol, with decreasing apolipoprotein C-II but without a commensurate decrease in RBP. The retinol levels had a positive correlation with apolipoprotein C-II in all or normolipidemic patients with diabetes and control subjects. The high retinol/RBP ratio implies that an excessive or free retinol possibly exists in NIDDM. An alternative metabolism of retinol is inferred to underlie NIDDM without direct influences of cholesterol or triglyceride themselves. PMID- 7485221 TI - Relative allergenicity of cow's milk and cow's milk-based formulas in an animal model. AB - Human infants fed cow's milk or cow's milk-based formula may become intolerant to milk proteins partially because of allergic reactions. However, the relative allergenicity of these diets has not been clearly determined at the intestinal level. In this study, the allergenicity of cow's milk and milk-based formula was evaluated by examining intestinal anaphylaxis caused by challenge with a milk protein fraction, beta-lactoglobulin (beta LG), in guinea pigs orally sensitized to these diets. Colonic segments were removed and challenged with beta LG. An antigen-induced, anaphylactically mediated elevation of transmural short circuit current, which is caused by net chloride (Cl-) secretion, was measured in Ussing chambers as an index of local immune responsiveness. After removal of the colon, all guinea pigs were challenged by intracardiac injection with beta LG to examine the onset of bronchospasm, a test for systemic anaphylaxis. Whereas colonic segments from all guinea pigs fed whole cow's milk responded to challenge with beta LG, segments from only 60% of animals fed cow's milk formula responded to the challenge. In addition, the responders in the latter group had a significantly lower magnitude of beta LG-induced Cl- secretion compared with animals sensitized to whole cow's milk. In parallel, bronchospasm developed in all guinea pigs fed whole cow's milk. In the group of animals fed cow's milk formula, bronchospasm developed only in those who responded to the intestinal challenge. On the basis of these results, cow's milk-based formula has less sensitizing power than whole cow's milk in our animal model, and our approach is effective in testing allergenicity at the intestinal level. PMID- 7485222 TI - Acute and chronic effects of nonsteroidal antiinflammatory drugs on glomerular filtration rate in elderly patients. AB - To assess the effects of nonsteroidal antiinflammatory drugs (NSAIDs) on glomerular filtration rate (GFR) in elderly patients with and without renal insufficiency, we conducted an open-label, randomized, prospective three-period cross-over study. Twenty-nine patients at least 65 years old were assigned to groups with preserved GFR (> 1.16 mL/s [70 mL/min]) or with renal insufficiency (GFR 0.50-1.16 mL/s [30-70 mL/min]). Patients received 800 mg ibuprofen three times daily, 20 mg piroxicam daily, or 200 mg sulindac twice daily for 1 month. Three-hour inulin and two-day creatinine clearances were measured before and after the first and last doses of NSAIDs. Ibuprofen, piroxicam, and sulindac decreased inulin clearance after single-doses in both groups of patients. In patients with renal insufficiency, creatinine clearance did not change after administration of ibuprofen for 1 month (0 +/- 0.06 mL/s, mean +/- standard error), but was decreased similarly with administration of either piroxicam or sulindac (-0.12 +/- 0.06 mL/s [-7.2 +/- 3.6 mL/min], P < 0.02). One patient with preserved GFR, but with other risk factors for NSAID-associated renal impairment, met our criteria for withdrawal by experiencing at least a 40 mumol/L (0.5 mg/dL) increase in serum creatinine above their baseline value. Our data indicate that NSAIDs do not adversely affect GFR in patients with preserved renal function unless they have another risk factor for NSAID-associated renal impairment. In contrast, patients with renal insufficiency may have significant chronic decrements in GFR with long-acting NSAIDs such as piroxicam and sulindac, but not with short-acting ibuprofen. Such patients should have renal function monitored while being treated with long-acting NSAIDs. PMID- 7485223 TI - Ketoacidosis-onset noninsulin dependent diabetes in Japanese subjects. AB - In 5 patients (2 women and 3 men, aged 16-36 years), diabetic ketoacidosis developed without precipitating illness. Pancreatic islet cell antibody was negative, and the duration of insulin dependency was shorter than 4 weeks. Hemoglobin A1c was < or = 6.3% for the mean period of 2.8 years thereafter, with diet therapy alone in 4 and with 5 mg glyburide in 1. Four were overweight before the development of diabetes, and 3 of them positive for family history of adult onset, non-ketotic diabetes. Frequency of human leukocyte antigen B61 was increased significantly in the patients. In a patient not previously overweight, family history of diabetes was negative, and human leukocyte antigen haplotypes common in insulin-dependent diabetes mellitus were accumulated. Serum immunoreactive insulin was within normal range or supranormal with normal glucose tolerance after recovery. The patients closely resemble black Americans with ketoacidosis-onset non-insulin dependent diabetes. PMID- 7485224 TI - Case report: thyrotropin-releasing hormone-induced myoclonus and tremor in a patient with Hashimoto's encephalopathy. AB - The authors investigated the possibility of a thyrotropin-releasing hormone related mechanism in a 43-year-old Japanese woman with Hashimoto's encephalopathy who experienced three relapses closely associated with the menstrual cycle. Her symptoms began at ovulation, worsened during the luteal phase, and improved during the menstruation phase. No abnormalities were found by brain magnetic resonance imaging and cerebral angiography. Intravenous administration of thyrotropin-releasing hormone induced symptoms of myoclonus and tremor similar to those observed during an exacerbation. The intensity and duration of involuntary movements induced by thyrotropin-releasing hormone were dose-dependent. The patient's symptoms were controlled effectively by thyroxine replacement therapy. On the basis of these findings, thyrotropin-releasing hormone may have an important role in Hashimoto's encephalopathy. PMID- 7485226 TI - Case report: subacute combined degeneration of the spinal cord from folate deficiency. AB - Subacute combined degeneration of the spinal cord is a rare neurologic complication of folate deficiency. Progressive gait disturbance, weakness, confusion, and depression developed in a 39-year-old man. He had taken phenobarbital for more than 2 years. He was bedbound, with new loss of position and vibration senses in the lower extremities. His hemoglobin was 2.9/dl, mean corpuscular volume 122 fl, vitamin B12 428 pg/ml, and folate 1 ng/ml. Peripheral blood and bone marrow showed megaloblastic anemia. Serum methylmalonic acid and homocysteine levels were consistent with folate deficiency, not B12 deficiency. Treatment with folate and packed erythrocytes resulted at 4 months in overall improvement, including walking. Position sense was restored, and vibration sense had become nearly normal. The authors found no cause for folate deficiency except phenobarbital. PMID- 7485225 TI - Case report: in situ hybridization for detection of inapparent infection with Chlamydia trachomatis in synovial tissue of a patient with Reiter's syndrome. AB - The authors have shown that protein antigens, RNA, and DNA from Chlamydia trachomatis are present in synovial tissues of patients with Reiter's syndrome (RS). However, those studies gave no insight into the host cell type involved or the precise tissue location of the bacteria. To address such issues, the authors developed an in situ hybridization system to detect chlamydia, and they used that system to examine synovial biopsies from a patient with RS and a patient without RS. The in situ system uses a previously described digoxigenin-labeled DNA probe that hybridizes with chlamydial 16S rRNA sequences in paraformaldehyde-fixed samples. Control studies with chlamydia-infected and uninfected HeLa cells confirmed that the in situ system is as sensitive as is direct fluorescence cytology for detection of the organism. Morphology of host and chlamydia cells is preserved after hybridization. Studies using synovial tissue from an osteoarthritis patient produced no in situ hybridization signal, but similar hybridization to tissue from a culture-/direct fluorescence cytology- negative RS patient had a strong intracellular signal for chlamydia within a subsynovial cell layer. These in situ hybridization results confirm the extensive presence of chlamydia in synovia and extend the authors' earlier observation that chlamydia RNA is present in the synovia of patients with RS. The data also confirm their electron microscopy studies, indicating that chlamydia are intracellular in synovial tissue, and they further show that infected host cells are located beneath the synovial lining. PMID- 7485227 TI - Review: new aspects of acute renal failure. PMID- 7485228 TI - Evidence that the APOE locus influences rate of disease progression in late onset familial Alzheimer's Disease but is not causative. AB - An association has been observed in several independent data sets between late onset Alzheimer's Disease (AD) and the APOE locus on chromosome 19. We have examined the genotype in family history positive (FHP) and family history negative (FHN) cases and find a distortion of the APOE allele frequencies in accord with previous studies. However, when we examined the allele distribution of the at-risk siblings of the FHP group we found an excess of the epsilon 4 allele which also differs significantly from historic controls but not from the affected siblings. The age distribution of the affected and unaffected siblings was similar, suggesting that the allelic frequency distortion in the unaffected siblings was not due to their being below the mean age of onset. Lod score linkage analysis, with age dependent onset and non-stringent specification of the genetic parameters, did not suggest linkage to the APOE locus. Furthermore, an analysis of variance of the age of disease free survival suggested that APOE genotype contributes a small fraction of the total variance indicating that the APOE locus is a poor predictor of disease free survival age within late onset families. One explanation for the age dependent association reported by other groups, and our results, is that the APOE locus enhances the rate of progression of the disease process in otherwise predisposed individuals and that variation at this locus is not able in and of itself to cause the disease. We suggest this hypothesis is compatible with the current literature regarding APOE and AD. PMID- 7485229 TI - SSCP analysis and sequencing of the human prion protein gene (PRNP) detects two different 24 bp deletions in an atypical Alzheimer's disease family. AB - Alzheimer's disease (AD) is a progressive, degenerative neurological disorder of the central nervous system. AD is the fourth leading cause of death in elderly persons 65 years or older in Western industrialized societies. The etiology of AD is unknown, but clinical, pathological, epidemiological, and molecular investigations suggest it is etiologically heterogeneous. Mutations in the amyloid protein are rare and segregate with the disease in a few early-onset familial AD (FAD) families. Similarities between AD and the unconventional viral (UCV) diseases, and between the amyloid and prion proteins, implicate the human prion protein gene (PRNP) as another candidate gene. Single strand conformation polymorphism (SSCP) analysis was used to screen for mutations at this locus in 82 AD patients from 54 families (30 FAD), vs. 39 age-matched controls. A 24-bp deletion around codon 68 that codes for one of five Gly-Pro rich octarepeats was identified in two affected sibs and one offspring of one late-onset FAD family. Two other affected sibs, three unaffected sibs, and three offspring from this family, in addition to one sporadic AD patient and three age-matched controls, were heterozygous for another octarepeat deletion located around codon 82. Two of the four affected sibs had features of PD, including one who was autopsy-verified AD and PD. Although these deletions were found infrequently in other AD patients and controls, they appear to be a rare polymorphism that is segregating in this FAD family. It does not appear that mutations at the PRNP locus are frequently associated with AD in this population. PMID- 7485230 TI - Autism, affective disorders, and social phobia. AB - The purpose of this study is to test the hypothesis that major affective and/or anxiety disorders are increased among relatives of autistic probands compared with controls. Among 36 families with an autistic child, 23 (64%) have a first degree relative diagnosed with major depressive disorder and 14 (39%) have a first degree relative diagnosed with social phobia. These rates are significantly greater than the 19% and 5%, respectively, found among 21 families with a child having a genetic condition, tuberous sclerosis complex, or a seizure disorder but no autism. The frequency of major depression among the 96 first degree relatives of autistic probands is 37.5% compared with 11.1% found among 45 relatives of control probands. The frequency of social phobia, 20.2%, is approximately 10 times more common than that found among the relatives of the control probands (2.4%). Elevated rates of both major depression and social phobia are found among parents and siblings in the families with an autistic child. Furthermore, 64% of parents affected with a major depression had the onset of the first depressive episode prior to the birth of the autistic child and all parents with social phobia had the onset of condition prior to the birth of the autistic child. Family patterns differ depending on the intellectual level of the autistic child; specifically, social phobia is significantly greater among the first degree relatives of non-retarded autistic probands than among relatives of individuals with autism and comorbid mental retardation.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7485231 TI - Why is the reproductive performance lower in Becker (BMD) as compared to limb girdle (LGMD) muscular dystrophy male patients? AB - We had previously reported that patients affected with BMD have a significantly reduced reproductive performance (f = 0.12) as compared to male LGMD patients of similar age and physical impairment (f = 0.98). In the present study parameters such as the socio-economic level, as well as psychosocial, intellectual, and psychiatric functionings could not explain the low fitness of BMD patients. The effect of genetic counseling, a greater difficulty in coping with the disease, and relating to women and/or a potential malfunction of reproductive physiology are discussed as possible causes. PMID- 7485232 TI - Becker and limb-girdle muscular dystrophies: a psychiatric and intellectual level comparative study. AB - There are some indications that Becker muscular dystrophy (BMD) might be related to mental disorders and mental retardation (MR). To investigate this question, we made a standardized psychiatric and intellectual level assessment of 22 BMD patients in comparison with 22 limb-girdle muscular dystrophy (LGMD) patients. There were not significant differences between the two groups. Twelve patients (54.5%) in each group received at least one lifetime psychiatric diagnosis, the most frequent being depressive disorders. The intelligence quotient means for BMD was 85.9 and 87.8 for LGMD. There was one case of mild MR among BMD patients and two cases among LGMD patients. PMID- 7485233 TI - Word-finding difficulties, verbal paraphasias, and verbal dyspraxia in ten individuals with fragile X syndrome. AB - Speech/language disorders are common in the fragile X syndrome. [Howard-Peebles, 1979: Am J Hom Genet 31:214-222; Renier et al., 1983: J Ment Defic Res 27:51-59; Sparks, 1984: Birth Defects and Speech-Language Disorders, pp. 39-43; Hanson et al., 1986: Am J Med Genet 23:195-206]. Verbal paraphasias have been considered a rare feature and word-finding difficulties have seldom been reported. Here we report on ten Brazilian patients who were evaluated for speech/language disturbances and found that word-finding difficulties were present in 50% of the cases, which is a slightly higher frequency than that of clear dyspraxia. We suggest, therefore, that word-finding difficulties and verbal dyspraxia can be a common feature within the spectrum of this syndrome. Additional speech findings are discussed. PMID- 7485234 TI - Clinical features of early onset, familial Alzheimer's disease linked to chromosome 14. AB - Early onset familial Alzheimer's disease (AD) has an autosomal dominant mode of inheritance. Two genes are responsible for the majority of cases of this subtype of AD. Mutations in the beta-amyloid precursor protein (beta APP) gene on chromosome 21 have been shown to completely cosegregate with the disease. We and others have previously described the clinical features of families with beta APP mutations at the codon 717 locus in an attempt to define the phenotype associated with a valine to isoleucine (Val-->Ile) or a valine to glycine (Val-->Gly) change. More recently, a second locus for very early onset disease has been localized to chromosome 14. The results of linkage studies in some families suggesting linkage to both chromosomes have been explained by the suggestion of a second (centromeric) locus on chromosome 21. Here we report the clinical features and genetic analysis of a British pedigree (F74) with early onset AD in which neither the beta APP locus nor any other chromosome 21 locus segregates with the disease, but in which good evidence is seen for linkage on the long arm of chromosome 14. In particular we report marker data suggesting that the chromosome 14 disease locus is close to D14S43 and D14S77. Given the likelihood that F74 represents a chromosome 14 linked family, we describe the clinical features and make a limited clinical comparison with the beta APP717 Val-->Ile and beta APP717 Val-->Gly encoded families that have been previously described.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7485235 TI - Chromosomal abnormalities in a psychiatric population. AB - Over a 3.5 year period of time, 345 patients hospitalized for psychiatric problems were evaluated cytogenetically. The patient population included 76% males and 94% children with a mean age of 12 years. The criteria for testing was an undiagnosed etiology for mental retardation and/or autism. Cytogenetic studies identified 11, or 3%, with abnormal karyotypes, including 4 fragile X positive individuals (2 males, 2 females), and 8 with chromosomal aneuploidy, rearrangements, or deletions. While individuals with chromosomal abnormalities do not demonstrate specific behavioral, psychiatric, or developmental problems relative to other psychiatric patients, our results demonstrate the need for an increased awareness to order chromosomal analysis and fragile X testing in those individuals who have combinations of behavioral/psychiatric, learning, communication, or cognitive disturbance. PMID- 7485236 TI - Genetic threshold hypothesis of neocortical spike-and-wave discharges in the rat: an animal model of petit mal epilepsy. AB - Neocortical high-voltage spike-and-wave discharges (HVS) in the rat are an animal model of petit mal epilepsy. Genetic analysis of total duration of HVS (s/12 hr) in reciprocal F1 and F2 hybrids of F344 and BN rats indicated that the phenotypic variability of HVS cannot be explained by a simple, monogenic Mendelian model. Biometrical analysis suggested the presence of additive, dominance, and sex linked-epistatic effects, buffering maternal influence, and heterosis. High correlation was observed between average duration (s/episode) and frequency of occurrence of spike-and-wave episodes (n/12 hr) in parental and segregating generations, indicating that common genes affect both duration and frequency of the spike-and-wave pattern. We propose that both genetic and developmental environmental factors control an underlying quantitative variable, which, above a certain threshold level, precipitates HVS discharges. These findings, together with the recent availability of rat DNA markers for total genome mapping, pave the way to the identification of genes that control the susceptibility of the brain to spike-and-wave discharges. PMID- 7485237 TI - Quantitative genetic analysis of injury liability in infants and toddlers. AB - A threshold model of latent liability was applied to infant and toddler twin data on total count of injuries sustained during the interval from birth to 36 months of age. A quantitative genetic analysis of estimated twin correlations in injury liability indicated strong genetic dominance effects, but no additive genetic variance was detected. Because interpretations involving overdominance have little research support, the results may be due to low order epistasis or other interaction effects. Boys had more injuries than girls, but this effect was found only for groups whose parents were prompted and questioned in detail about their children's injuries. Activity and impulsivity are two behavioral predictors of childhood injury, and the results are discussed in relation to animal research on infant and adult activity levels, and impulsivity in adult humans. Genetic epidemiological approaches to childhood injury should aid in targeting higher risk children for preventive intervention. PMID- 7485238 TI - Schizoaffective psychoses: genetical clues to classification. AB - The diagnostic classification of schizoaffective psychoses has varied much since Kasanin introduced the concept in 1933. The various classifications have agreed that schizoaffective psychoses present a combination of schizophreniform and affective symptoms, but the diagnostic criteria differ as to the number, quality, and time sequence of the symptoms even in recent classifications like RDC, DSM III-R, and ICD-10. The classifications are syndromatical, and the etiology of the schizoaffective psychoses is still undetermined apart from evidence for a strong genetic factor. Results from family, twin, and adoption studies are divergent, but all the same, support a separate classification of broadly defined schizoaffective psychoses as possibly being phenotypical variations or expressions of genetic interforms between schizophrenia and affective psychoses. PMID- 7485239 TI - Exclusion of linkage between alcoholism and the MNS blood group region on chromosome 4q in multiplex families. AB - Polymorphic DNA markers on the long arm of chromosome 4 were used to examine linkage to alcoholism in 20 multiplex pedigrees. Fifteen loci were determined for 124 individuals. Lod scores were calculated assuming both dominant and recessive disease modes of inheritance, utilizing incidence data by age and gender that allow for correction for variable age of onset and frequency of the disorder by gender. Under the assumption that alcoholism is homogeneous in this set of pedigrees, and that a recessive mode with age and gender correction is the most appropriate, the total lod scores for all families combined were uniformly lower than -2.0. This suggests an absence of linkage between the putative alcoholism susceptibility gene and markers in the region of the MNS blood group (4q28-31), a region for which we had previously found suggestive evidence of linkage to alcoholism. The 100 cM span of chromosome 4 studied includes the class I alcohol dehydrogenase (ADH) loci. Using the recessive mode, no evidence for linkage to alcoholism was found for the markers tested, which spanned almost the entire long arm of chromosome 4. Under the dominant mode, no evidence for linkage could be found for several of the markers. PMID- 7485240 TI - Lafora disease is not linked to the Unverricht-Lundborg locus. AB - Lafora disease and Unverricht-Lundborg disease are two forms of progressive myoclonus epilepsies (PME). Recently the gene for Unverricht-Lundborg disease (EPM1) was mapped to chromosome 21q22.3. Using three highly polymorphic DNA markers (D21S212, PFKL, and D21S171) which flank the EPM1 locus, we performed linkage analysis to investigate whether or not the EPM1 gene is also implicated in Lafora disease. Linkage was excluded in three North-African pedigrees each comprising at least two affected individuals. This result suggests that differential diagnosis of Lafora disease and Unverricht-Lundborg disease may be facilitated by molecular genetic analysis. PMID- 7485241 TI - No association between schizophrenia and polymorphisms within the genes for debrisoquine 4-hydroxylase (CYP2D6) and the dopamine transporter (DAT). AB - It has been suggested that the cytochrome P450 mono-oxygenase, debrisoquine 4 hydroxylase, is involved in the catabolism and processing of neurotransmitters subsequent to their reuptake into target cells. It is also thought to be related to the dopamine transporter that acts to take released dopamine back up into presynaptic terminals. The present study used the association approach to test the hypothesis that mutations in the genes for debrisoquine 4-hydroxylase (CYP2D6) and the dopamine transporter (DAT) confer susceptibility to schizophrenia. There were no differences in allele or genotype frequencies between patients and controls in the mutations causing the poor metaboliser phenotype in CYP2D6. In addition there was no association found between schizophrenia and a 48bp repeat within the 3' untranslated region of DAT. PMID- 7485243 TI - Normal CAG and CCG repeats in the Huntington's disease genes of Parkinson's disease patients. AB - The clinical features of Parkinson's disease, particularly rigidity and bradykinesia and occasionally tremor, are seen in juvenile-onset Huntington's disease. Therefore, the CAG and CCG repeats in the Huntington's disease gene were investigated in 45 Parkinson's disease patients and compared to 40 control individuals. All of the Parkinson's disease chromosomes fell within the normal size ranges. In addition, the distributions of the two repeats in the Parkinson's disease patients did not differ significantly from those of the control population. Therefore, abnormalities of these trinucleotide repeats in the Huntington's disease gene are not likely to contribute to the pathogenesis of Parkinson's disease. PMID- 7485244 TI - Role of genetic factors in depression based on studies of Tourette syndrome and ADHD probands and their relatives. AB - Tourette syndrome (TS) is a common, neuropsychiatric disorder which has many similarities to attention deficit hyperactivity disorder (ADHD). TS probands have a high frequency of a variety of behavioral disorders including depression. The depression may be due to a pleiotrophic effect of the Gts genes, proband ascertainment bias, or a result of coping with the chronic tics. To distinguish between these hypotheses we examined the responses to 17 Diagnostic Interview Schedule questions to evaluate the 9 DSM-III-R criteria for major depressive episode in 1,080 adults consisting of TS and ADHD probands, their relatives and controls. Using a Bonferonni corrected p there was a significant progressive increase in 16 of 17 depressive symptoms and for a life time history of a major depressive episode in groups with increased genetic loading for Gts genes. Similar trends were seen in the small number of ADHD probands and their relatives. There was also a significant increase for these variables in non proband TS relatives versus non-TS relatives, indicating the association of depression with Gts genes was not due to ascertainment bias or the inappropriate choice of controls. Multiple linear regression analysis indicated that obsessive compulsive behaviors, sex, ADHD, drug abuse, and age all showed a more significant effect on depressive symptoms than the number of tics. The presence or absence of TS in the relatives had a much greater effect on risk for depression than the presence or absence of an episode of major depression in the proband. These results are consistent with the hypothesis that Gts and ADHD genes play a major role in depression. PMID- 7485242 TI - Genetic linkage analysis of schizophrenia using chromosome 11q13-24 markers in Israeli pedigrees. AB - It is generally agreed that there is a genetic component in the etiology of schizophrenia which may be tested by the application of linkage analysis to multiply-affected families. One genetic region of interest is the long arm of chromosome 11 because of previously reported associations of genetic variation in this region with schizophrenia, and because of the fact that it contains the locus for the dopamine D2 receptor gene. In this study we have examined the segregation of schizophrenia with microsatellite dinucleotide repeat DNA markers along chromosome 11q in 5 Israeli families multiply-affected for schizophrenia. The hypothesis of linkage under genetic homogeneity of causation was tested under a number of genetic models. Linkage analysis provided no evidence for significant causal mutations within the region bounded by INT and D11S420 on chromosome 11q. It is still possible, however, that a gene of major effect exists in this region, either with low penetrance or with heterogeneity. PMID- 7485246 TI - Relation of schizophrenia and panic disorder: evidence from a controlled family study. AB - The intention of this controlled family study was to evaluate reasons for comorbidity of schizophrenia and panic disorder. Observed rates of psychiatric disorders in first-degree relatives of patients and of controls were compared with rates predicted by possible hypotheses explaining comorbidity. The sample consisted of 59 patients with schizophrenia (including seven with schizophreniform disorder), 54 patients with panic disorder (with or without agoraphobia), 29 comorbid patients with lifetime diagnoses of panic disorder and schizophrenia (or schizophreniform disorder, 2 patients) and 109 controls, and their 1068 first-degree relatives. Information from clinical performance, clinical and structured interviews, and from family history was joined to establish DSM-III-R diagnoses in patients and relatives. As expected, schizophrenia and panic disorder were found to be familial. The hypothesis, that the familial load for primary panic disorders distinguished schizophrenics (4.3%) and controls (0.9%), could be verified (P < 0.01); the familial aggregation for primary panic disorders did not distinguish schizophrenics and subjects with panic disorder. However, the risk for schizophrenia was not enhanced in relatives of patients with panic disorder (0%) in comparison to controls (0.3%, P > 0.05). The observed familial aggregation pattern of psychiatric disorders in relatives of schizophrenics, panic patients, comorbid patients, and controls refers to an etiological relation of schizophrenia and panic disorder, or at least to a relationship of subgroups of these disorders. PMID- 7485245 TI - Preliminary evidence for an association of a dinucleotide repeat polymorphism at the MAOA gene with early onset alcoholism/substance abuse. AB - An association between the liability to early onset alcoholism/substance abuse and a recently discovered dinucleotide repeat length polymorphism at the MAOA gene (MAOCA-1) was examined using polymerase chain reaction (PCR). A significant correlation between the presence/absence of the disorder and the length of the MAOCA-1 repeat was found in males, but not females, with "long" alleles (repeat length above 115 bp) associated with both increased risk for the disorder and lower age of onset of substance abuse. These preliminary data suggest that further exploration of the relationship between the MAOA gene and behavioral traits in an expanded sample is warranted. PMID- 7485247 TI - Constitutive heterochromatin of chromosome 1 and Duffy blood group alleles in schizophrenia. AB - Cytogenetic analysis was carried out in unrelated schizophrenic patients, unrelated controls and patients and family members in multiplex families. The size distribution of chromosome 1 heterochromatic region (1qH, C-band variants) among 21 unrelated schizophrenic patients was different from that found in a group of 46 controls. The patient group had 1qH variants of smaller size than the control group (P < 0.01). Incubation of phytohemagglutinin-treated blood lymphocytes with 5-azacytidine (which causes decondensation and extension of the heterochromatin) led to a lesser degree of heterochromatin decondensation in a group of patients than in the controls (7 schizophrenic, 9 controls, P < 0.01). The distribution of phenotypes of Duffy blood group system [whose locus is linked to the 1qH region (Donahue et al.: Proc Natl Acad Sci USA 61:949-955, 1968; Rouleau et al.: Genomics 7:313-318, 1990)] among 28 schizophrenic patients was also different from that in the general population. Cosegregation of schizophrenia with a 1qH (C-band) variant and Duffy blood group allele was observed in one of six multiplex families. The overall results suggest that alterations within the Duffy/1qH region are involved in schizophrenia in some cases. This region contains the locus of D5 dopamine receptor pseudogene 2 (1q21.1), which is transcribed in normal lymphocytes (Takahashi et al.: FEBS Lett 314:23-25, 1992). PMID- 7485248 TI - Linkage studies on chromosome 22 in familial schizophrenia. AB - As part of a systematic search for a major genetic locus for schizophrenia we have examined chromosome 22 using 14 highly polymorphic markers in 23 disease pedigrees. The markers were distributed at an average distance of 6.6 cM, covering 70-80% of the chromosome. We analyzed the data by the lod score method using five plausible genetic models ranging from dominant to recessive, after testing the power of our sample under the same genetic parameters. The most positive lod score found was 1.51 under a recessive model for the marker D22S278, which is insufficient to conclude linkage. However, an excess of shared alleles in affected siblings (P < .01) was found for both D22S278 and D22S283. For D22S278, the A statistic was equal to the lod score (1.51) and therefore did not provide additional evidence for linkage allowing for heterogeneity, but the Liang statistic was more significant (P = .002). Our results suggest the possibility that the region around D22S278 and D22S283 contains a gene which contributes to the aetiology of schizophrenia. PMID- 7485249 TI - Two single base polymorphisms in introns 41 and 16 of the NF1 gene. AB - We have characterized two intragenic polymorphisms in the neurofibromatosis type 1 (NF1) gene by direct sequencing of PCR products. The variants for these polymorphisms were initially detected on Hydrolink gels. One of the polymorphisms involves a G to A transition in intron 41 at the 28th base upstream of exon 42 with an observed 'G'/'A' heterozygosity of 0.42. The other polymorphism is a T to C transition in intron 16 at the 16th base upstream of exon 17 with an observed 'T'/'C' heterozygosity of 0.09. In combination with other documented polymorphisms in the NF1 gene, these variants should assist in genetic analysis of NF1 families. PMID- 7485251 TI - Schizophrenia: gender, family risk, and plasma homovanillic acid. AB - Plasma homovanillic acid concentration was assessed in 60 young schizophrenic patients, with and without first-degree relatives with schizophrenia, before treatment, and 3 days after starting haloperidol treatment. The baseline concentration of homovanillic acid in plasma was no different in the two groups before treatment; it was, however, significantly higher in the patients with relatives than in those without relatives diagnosed of schizophrenia after 3 days of haloperidol treatment. PMID- 7485250 TI - Mutation scan of the D1 dopamine receptor gene in 22 cases of bipolar I disorder. AB - In a previous study [Coon et al., Am J Hum Genet 52:1234-1249, 1993], we found suggestive evidence of linkage between manic-depressive illness (MDI) in eight multiplex pedigrees and D5S62, a DNA marker mapping to the telomeric region of 5q. As the D1 dopamine receptor gene (DRD1) maps to this region and as alterations in dopaminergic neurotransmission have been indirectly implicated in the pathogenesis of MDI, we directly searched for mutations in the coding region of the DRD1 gene in 22 unrelated cases of bipolar I (BPI) disorder derived from multiplex families. Using single strand conformation polymorphism (SSCP) analysis, we did not observe any abnormal SSCP variants in the BPI cases that differed from controls. PMID- 7485252 TI - Mapping of the serotonin 5-HT1D beta autoreceptor gene on chromosome 6 and direct analysis for sequence variants. AB - Abnormal brain serotonin function may be characteristic of several neuropsychiatric disorders. Thus, it is important to identify polymorphic genes and screen for functional variants at loci coding for genes that control normal serotonin functions. 5-HT1D beta is a terminal serotonin autoreceptor which may play a role in regulating serotonin synthesis and release. Using an SSCP technique we screened for 5-HT1D beta coding sequence variants in psychiatrically interviewed populations, which included controls, alcoholics, and alcoholic arsonists and alcoholic violent offenders with low CSF concentrations of the main serotonin metabolite 5-HIAA. A common polymorphism was identified in the 5-HT1D beta gene with allele frequencies of 0.72 and 0.28. The SSCP variant was caused by a silent G to C substitution at nucleotide 861 of the coding region. This polymorphism could also be detected as a HincII RFLP of amplified DNA. DNAs from informative CEPH families were typed for the HincII RFLP and analyzed with respect to 20 linked markers on chromosome 6. Multipoint analysis placed the 5 HT1D beta receptor gene between markers D6S286 and D6S275. A maximum two-point lod score of 10.90 was obtained to D6S26, which had been previously localized on 6q14-15. Chromosomal aberrations involving this region have been previously shown to cause retinal anomalies, developmental delay, and abnormal brain development. This region also contains the gene for North Carolina-type macular dystrophy. PMID- 7485254 TI - Screening the dopamine D1 receptor gene in 131 schizophrenics and eight alcoholics: identification of polymorphisms but lack of functionally significant sequence changes. AB - To determine whether mutations in the D1 dopamine receptor (D1 DR) gene are associated with schizophrenia, the coding sequence was examined in 106 Caucasian, 11 African-American, 8 Asian, and 6 Native American patients. Approximately 350 kb of genomic sequence was screened by dideoxy fingerprinting, a method related to single strand conformational polymorphism (SSCP) analysis that detects virtually 100% of sequence changes [Sarkar et al., 1992: Genomics 13:441-443; Liu and Sommer, 1994: PCR Methods and Applications 4:97-108]. One polymorphism was identified in Asians and one in Caucasians, but neither altered the amino acid sequence (Leu66, and Ser421, respectively). In addition, a previously reported polymorphism in the 5' untranslated region of exon 2 at bp -48 was found to be common, with an allele frequency of approximately 40% in Caucasians of Western European descent. Based on the fact that no sequence changes of likely functional significance were identified, these data suggest that mutations affecting the structure of the D1 dopamine receptor protein are uncommon and are unlikely to contribute significantly to the genetic predisposition to schizophrenia. The D1 DR gene also was examined in eight alcoholics, including 3 African-Americans and 1 Native American, but no sequence changes were identified. PMID- 7485253 TI - Mapping of the serotonin 5-HT1D alpha autoreceptor gene (HTR1D) on chromosome 1 using a silent polymorphism in the coding region. PMID- 7485255 TI - Follow-up report of potential linkage for schizophrenia on chromosome 22q: Part 3. PMID- 7485256 TI - Further delineation of brain anomalies in velo-cardio-facial syndrome. PMID- 7485257 TI - Allelic association at the D14S43 locus in early onset Alzheimer's disease. French Alzheimer's Disease Collaborative Study Group. AB - The D14S43 marker is closely linked to the major gene for early onset autosomal dominant Alzheimer's disease on chromosome 14. Allelic frequencies at the D14S43 locus were compared in 113 familial and isolated cases of early onset Alzheimer's disease (< 60 years of age at onset) (EOAD) and 109 unaffected individuals of the same geographic origin. Allele 7 was significantly (P = 0.033) more frequent in type 1 EOAD patients (13.2%), defined by the presence of at least another first degree relative with EOAD, than in controls (4.1%). Since an autosomal dominant gene is probably responsible for type 1 patients, allelic association may reflect linkage disequilibrium at the D14S43 locus. This would mean that some patients share a common ancestral mutation. However, since multiple tests were carried out, this result must be interpreted with caution, and needs confirmation in an independent sample. PMID- 7485258 TI - Linkage studies of bipolar disorder in the region of the Darier's disease gene on chromosome 12q23-24.1. AB - We have recently described a family in which there is cosegregation of major affective disorder with Darier's disease and have mapped this autosomal dominant skin disorder to 12q23-q24.1. This has provided an interesting candidate region for genetic studies of bipolar disorder. We have studied the segregation of seven markers spanning the Darier's disease locus in 45 bipolar disorder pedigrees and found modest evidence in support of linkage under heterogeneity for 5 of these markers. Nonparametric analyses were suggestive of linkage with a marker at the gene encoding a secretory form of phospholipase A2. Our sample has relatively low power to detect linkage under heterogeneity and independent researchers should examine markers from this region in further samples of bipolar pedigrees. PMID- 7485259 TI - Association and linkage studies of the TAQI A1 allele at the dopamine D2 receptor gene in samples of female and male alcoholics. AB - To address the controversy surrounding DRD2 and alcoholism, we performed linkage and association studies utilizing alcoholic men from high-density families largely uncontaminated by other psychopathology and female alcoholics for whom secondary drug dependence (averaging 10 years later onset) was a prominent feature. The males and females were combined for a total of 52 alcoholics, and compared to 30 controls screened for the absence of alcoholism and other psychopathology, revealing a significant association between the frequency of the TaqI A1 allele and alcoholism. However, linkage and family-based association studies conducted on 20 families of male alcoholics found no evidence for association or linkage between Taq A and alcoholism. The results of our population-based association study, placed in the context of the literature, suggest that minimizing psychopathology in control groups is probably a more important explanation for divergent results than either sampling error or population stratification. When combined with the complete lack of within-family evidence, we concluded that the association, while not appearing to be artifactual, is not specific to the alcoholism phenotype, per se. PMID- 7485263 TI - Complex segregation analyses of old order Amish families ascertained through bipolar I individuals. AB - Specific genetic hypotheses about the mode of transmission of bipolar affective disorders were examined by performing complex segregation analyses of Old Order Amish families. The analyses were performed on 1) the total set of 42 families including 689 relatives, 2) a subset of 19 families consisting of those kindreds sharing common ancestors within three generations that contained 333 relatives, and 3) a subset of 23 more distantly related families with 356 relatives. When all 42 families were included in the analyses, the specific mode of transmission that could be distinguished was dependent upon the diagnostic scheme used in the analysis. An autosomal dominant mode of inheritance could be rejected when relatives with bipolar I, atypical bipolar, major depressive disorder, and hypomania were included as affected. When analyses included only the subset of families more closely related, an autosomal dominant inheritance model was found to be consistent with transmission of BP I disorder. It was not possible to distinguish between other transmission models with broader diagnostic schemes in this subset of families. Finally, results of analyses on the subset of more distantly related families suggest that there is a significant proportion of Old Order Amish families in which the genetic factors contributing to the expression of bipolar illness are either polygenic or oligogenic. PMID- 7485260 TI - What can the DRD2/alcoholism story teach us about association studies in psychiatric genetics? PMID- 7485262 TI - Parents and collateral relatives of children with pervasive developmental disorders: a family history study. AB - The objective of this study was to see whether, using the family history method, the risk for pervasive developmental disorder (PDD), cognitive impairments, and other psychiatric symptoms is greater in the parents and collateral relatives of probands with PDD compared to a control group. A semistructured family history interview was carried out with the parents of 52 probands with PDD and 33 parents of controls. Rates of cognitive impairments and psychiatric problems were not found more frequently in parents or relatives of PDD probands compared to relatives of controls, but four cases of PDD were reported among the extended families of the PDD probands. The relatives with PDD were related to the probands through the maternal line, possibly suggesting some form of maternal influence on inheritance or reduced penetrance in females with the PDD genotype. PMID- 7485261 TI - Autism and genetics: clinical approach and association study with two markers of HRAS gene. AB - Twin studies and familial aggregation studies indicate that genetic factors could play a role in infantile autism. In an earlier study, we identified a possible positive association between autism and a c-Harvey-ras (HRAS) oncogene marker at the 3' end of the coding region. In an attempt to confirm this finding, we studied a larger population, well-characterized clinically and genetically. We report a positive association between autism and two HRAS markers, the 3' marker used in the initial study and an additional marker in exon 1. PMID- 7485264 TI - Linkage analysis of chromosome 22q12-13 in a United Kingdom/Icelandic sample of 23 multiplex schizophrenia families. AB - A possible linkage to a genetic subtype of schizophrenia and related disorders has been reported on the long arm of chromosome 22 at q12-13. (Pulver et al., 1994: Am J Med Genet 54:36-43; Coon et al., 1994: Am J Med Genet 54:72-79; Pulver et al., 1994: Am J Med Genet 54:44-50). However formal statistical tests in a combined sample could not reject homogeneity and prove that there was a linked subgroup of families. We have studied 23 schizophrenia pedigrees to test whether some multiplex schizophrenia families may be linked to the microsatellite markers D22S274 and D22S283 which span the 22q12-13 region. Two point followed by multipoint lod and non-parametric linkage analyses under the assumption of heterogeneity provided no evidence for linkage over the relevant region. PMID- 7485266 TI - Confirmation of locus heterogeneity in the pure form of familial spastic paraplegia. AB - Familial spastic paraplegia (FSP), characterized by progressive spasticity of the lower extremities, is in its "pure" form generally of autosomal dominant inheritance pattern. Hazan et al. [Nat Genet 5:163-167, 1993] reported tight linkage of a large FSP family to the highly polymorphic microsatellite marker D14S269 with z (theta) = 8.49 at theta = 0.00 They further demonstrated evidence for locus heterogeneity when they showed that 2 FSP families were unlinked to this region. We have subsequently studied 4 FSP families (3 American, one British) and excluded the disease locus in these families for approximately 30 cM on either side of D14S269, thereby confirming evidence for locus heterogeneity within the spastic paraplegia diagnostic classification. PMID- 7485265 TI - Fragile X syndrome: discordant levels of CGG repeat mosaicism in two brothers. AB - Fragile X syndrome is associated with an unstable repeated CGG trinucleotide sequence in the 5' untranslated region of the FMR-1 gene. A significant number of individuals with a mild or atypical presentation are mosaics for the CGG expansion. We report a family with two brothers. The proband had severe mental retardation as well as most of the Fragile X syndrome stigmata, whereas his brother shows only mild learning difficulties. Both inherited a 80 x CGG trinucleotide premutation from the mother. They were negative for the FRAXA fragile site in over 100 metaphases examined. Flanking markers verified that both had inherited the same FMR-1 allele and Xq27-q28 flanking sequences from the mother. The methylation status of the brothers indicated active FMR-1 transcription as determined by using StB12.3/EcoRI + Eagl blots. CGG size or methylation mosaicism was not apparent from Southern blots. Polymerase chain reaction and chemiluminescent detection identified that both brothers had different degrees of mosaicism for the CGG expansion. Large expansions amounting to 70% of the total were visible in the proband, whereas less than 5% of the signal was larger than the premutation in his mildly affected brother. These findings suggest that mosaicism may be responsible for some of the variation in penetrance in this disorder. PMID- 7485267 TI - 17q inversion involving the neurofibromatosis type one locus in a family with neurofibromatosis type one. AB - We report a family with a paracentric inversion of the long arm of chromosome 17 [inv(17)(q11.2q25.1)] and neurofibromatosis type one (NF1). The family was ascertained because of NF1 and multiple miscarriages. Fluorescence in situ hybridization using cosmid probes from opposite ends of the NF1 gene confirmed that the inversion disrupts the gene. Using field inversion gel electrophoresis we have found that the inversion separates cDNA probes FB5D and AE25, which are normally adjacent to one another in the NF1 gene. This is the third published report of a gross chromosomal rearrangement responsible for NF1. The features in this family are typical for NF1, and are not unusually severe. PMID- 7485269 TI - No evidence for allelic association between bipolar disorder and monoamine oxidase A gene polymorphisms. AB - We have tested the hypothesis that DNA markers in the MAOA gene show allelic association with bipolar affective disorder. Eighty-four unrelated Caucasian patients with DSM III-R bipolar disorder and 84 Caucasian controls were typed for three markers in MAOA: a dinucleotide repeat in intron 2, a VNTR in intron 1, and an Fnu4HI RFLP in exon 8. No evidence for allelic association was observed between any of the markers and bipolar disorder. PMID- 7485268 TI - Adrenocorticotropin receptor/melanocortin receptor-2 maps within a reported susceptibility region for bipolar illness on chromosome 18. AB - We have examined the possible linkage of adrenocorticotropin receptor/melanocortin receptor-2 (ACTHR/MC-2) to a reported putative susceptibility locus for bipolar illness (BP) in 20 affected pedigrees. Initially, allelic variants of the gene were identified by polymerase chain reaction-single stranded conformation polymorphism (PCR-SSCP) and the gene was genetically mapped using both the Centre d'Etudes du Polymorphisme Humain (CEPH) pedigrees and the BP pedigrees used in this study. We found that the ACTHR/MC-2 gene maps between D18S53 and D18S66. These loci span a region of chromosome 18 which, in a previous study [Berrettini et al.: Proc Natl Acad Sci USA 91:5918 5921, 1994) revealed a putative predisposing locus to BP through nonparametric methods of linkage analysis. Linkage of ACTHR/MC-2 to BP was not demonstrable under parametric and nonparametric methods of analyses, although affected sib pair (ASP) method revealed an increase in allele sharing among ill individuals, P = 0.023. Since this receptor is within a potential linkage region, ACTHR/MC-2 could be considered a candidate gene for BP. PMID- 7485270 TI - Evidence for a genetic association between alleles of monoamine oxidase A gene and bipolar affective disorder. AB - We present evidence of a genetic association between bipolar disorder and alleles at 3 monoamine oxidase A (MAOA) markers, but not with alleles of a monoamine oxidase B (MAOB) polymorphism. The 3 MAOA markers, including one associated with low MAOA activity, show strong allelic association with each other but surprisingly not with MAOB. Our results are significant only for females, though the number of males in our sample is too small to draw any definite conclusions. Our data is consistent with recent reports of reduced MAOA activity in patients with abnormal behavioral phenotypes. The strength of the association is weak, but significant, which suggests that alleles at the MAOA locus contribute to susceptibility to bipolar disorder rather than being a major determinant. PMID- 7485271 TI - Occurrence of the Cys311 DRD2 variant in a pedigree multiply affected with panic disorder. AB - Following the detection of the rare DRD2 codon 311 variant (Ser-->Cys) in an affected member from a large, multiply affected panic disorder family, we investigated the occurrence of this variant in other family members. The variant occurred in both affected and unaffected individuals. Further screening in panic disorder sib pairs unrelated to this family failed to detect the Cys311 variant. Our data suggests that this variant has no pathogenic role in panic disorder. PMID- 7485272 TI - Chromosomal locations and modes of action of genes of the retinoid (vitamin A) system support their involvement in the etiology of schizophrenia. AB - Vitamin A (retinoid), an essential nutrient for fetal and subsequent mammalian development, is involved in gene expression, cell differentiation, proliferation, migration, and death. Retinoic acid (RA) the morphogenic derivative of vitamin A is highly teratogenic. In humans retinoid excess or deficit can result in brain anomalies and psychosis. This review discusses chromosomal loci of genes that control the retinoid cascade in relation to some candidate genes in schizophrenia. The paper relates the knowledge about the transport, delivery, and action of retinoids to what is presently known about the pathology of schizophrenia, with particular reference to the dopamine hypothesis, neurotransmitters, the glutamate hypothesis, retinitis pigmentosa, dermatologic disorders, and craniofacial anomalies. PMID- 7485273 TI - Applying systems theory. PMID- 7485274 TI - Taking on the TV shows. PMID- 7485275 TI - Burnout. Why do we blame the nurse? PMID- 7485276 TI - What to expect from your malpractice attorney. PMID- 7485277 TI - Performing foot screening for diabetic patients. PMID- 7485278 TI - Clinical snapshot: portal hypertension. PMID- 7485279 TI - Assessing fluid and electrolyte balance in elders. PMID- 7485280 TI - Ethical issues: withdrawing life support: who decides? PMID- 7485281 TI - Emergency! Renal calculi. PMID- 7485282 TI - When a baby dies. PMID- 7485283 TI - Your license to practice: variations on a theme. PMID- 7485284 TI - Silent voices. PMID- 7485285 TI - Management and outcome of pregnant women with interstitial and restrictive lung disease. AB - OBJECTIVE: Our purpose was to describe our experience with management and outcome of restrictive lung disease in pregnancy. STUDY DESIGN: Between 1981 and 1994 nine pregnant women with interstitial and restrictive lung disease were prospectively managed and delivered at the University of Washington Medical Center. RESULTS: Three patients had severe disease, characterized by vital capacity < or = 1.5 L (50% predicted) or diffusing capacity < or = 50% predicted. Five patients had exercise-induced oxygen desaturation, and four required supplemental oxygen. Five patients required corticosteroids. One patient had an adverse outcome; she was delivered at 31 weeks and required mechanical ventilation for 72 hours. All other patients were delivered at or beyond 36 weeks with no adverse intrapartum or postpartum complications. All babies were at or above the 30th percentile for growth. CONCLUSIONS: Restrictive lung disease is tolerated in pregnancy. Exercise intolerance is common and patients may require early oxygen supplementation. Adequate fetal growth can be achieved. PMID- 7485286 TI - Delivery of the nonvertex second twin: breech extraction versus external cephalic version. AB - OBJECTIVE: Our purpose was to compare the maternal and perinatal outcomes of twin gestations in which the nonvertex second twin was delivered by total breech extraction versus those delivered by external cephalic version. STUDY DESIGN: The intrapartum courses of 284 consecutive twin gestations were analyzed retrospectively. Once those with actual birth weight < 600 gm, unrecognized multifetal pregnancy, multiple congenital anomalies, cesarean delivery, and/or antepartum intrauterine fetal death were excluded, 23 mothers were delivered by total breech extraction and 21 underwent external cephalic version. RESULTS: The two groups were similar for mean (+/- SD) maternal age, gravidity, parity, gestational age at delivery, ultrasonographic estimate of birth weight for twin B, incidence of breech or transverse presentation for the second fetus, and actual birth weight of the first or second newborn. Suspected fetal distress that led to cesarean delivery occurred significantly more often in parturients who underwent attempted external version (4/21) than total breech extraction (0/23, p = 0.04). The incidence of eventual abdominal delivery was also significantly higher in patients who underwent attempted external cephalic version (10/21) rather than breech extraction (1/23, p = 0.001). For twin B the occurrence of low Apgar scores at 1 minute was significantly higher for infants after attempted external version (7/21) rather than breech extraction (1/23, p = 0.02), but the mean pH, number with Apgar scores < 7 at 5 minutes, and number of neonatal intensive care unit admissions were similar. No perinatal traumatic injury occurred in either group. CONCLUSION: On the basis of our experience, total breech extraction of the nonvertex second twin is preferable to external cephalic version because it appears to be associated with a significantly lower incidence of fetal distress and abdominal delivery with comparable neonatal outcome. PMID- 7485287 TI - Comparison of intrapartum electronic fetal heart rate monitoring versus intermittent auscultation in detecting fetal acidemia at birth. AB - OBJECTIVE: Our purpose was to compare continuous intrapartum electronic fetal heart rate monitoring with intermittent auscultation for detecting fetal acidemia at birth. STUDY DESIGN: Data from a previously published randomized trial of electronic fetal heart rate monitoring versus intermittent auscultation were analyzed to identify any differences between the two methods in detecting fetal acidemia at birth. Fetal acidemia at birth was defined as the presence of cord blood arterial pH < 7.15. RESULTS: A total of 1419 patients with umbilical cord blood acid-base measurements were identified, 739 in the electronic FHR monitoring group and 680 in the auscultation group. Electronic FHR monitoring had significantly better sensitivity (97% vs 34%, p < 0.001), lower specificity (84% vs 91%, p < 0.001), higher positive predictive value (37% vs 22%, p < 0.05), and higher negative predictive value (99.5% vs 95%, p < 0.001) in detecting fetal acidemia at birth. In addition, electronic FHR monitoring was significantly better in detecting all types of acidemia: metabolic (95.5% vs 26.5%, p < 0.001), mixed (95% vs 37.5%, p < 0.001), and respiratory (100% vs 41.5%, p < 0.001). CONCLUSION: These data suggest that electronic FHR monitoring is superior to intermittent auscultation in detecting fetal acidemia at birth. PMID- 7485288 TI - Is the 15-in situ clone protocol necessary to detect amniotic fluid mosaicism? AB - OBJECTIVE: Our purpose was to evaluate the 15-clone analysis for detecting amniotic fluid mosaicism by the in situ method. STUDY DESIGN: A 10-year review was performed of all amniotic fluid mosaicism cases at two institutions using the in situ method exclusively, with sequential clonal analysis to determine the first and second clone in which the abnormal cell line occurred. RESULTS: Of the 28,497 amniotic fluid samples, 73 met criteria for amniotic fluid mosaicism by in situ method (0.26%). There were 54 cases (0.19%) with potential clinical significance (23 autosome and 31 sex chromosome mosaicism); 49 of the 54 cases (89%) were detected in the first six clones, including 22 of 23 involving autosomes and 27 of 31 involving sex chromosomes. In one of the six cases detected after clone 6 (46,XX/47,XX,+21) the mosaic cell line was present in 20% of the clones analyzed and was followed by a voluntary termination of the pregnancy. In the other five cases amniotic fluid mosaicism was present in < 20% of the clones; these included one case of 46,XX/47,XX+mar (15% amniotic fluid mosaicism, voluntary termination of pregnancy), two cases of 45,X/46,XY (10% to 12% amniotic fluid mosaicism, both normal at birth), and two cases of 45,X/46,XX (8% amniotic fluid mosaicism, lost to follow-up; 12% amniotic fluid mosaicism, voluntary termination of pregnancy). By limiting the analysis to six clones, approximately 20% of analysis time could be saved per case, but one autosomal amniotic fluid mosaicism case per 10,000 samples could potentially be missed. CONCLUSION: Reducing the number of clones analyzed by in situ method could result in increased efficiency, decreased costs, and minimal loss of sensitivity. PMID- 7485289 TI - Role of prenatal ultrasonography in women with positive screen for Down syndrome on the basis of maternal serum markers. AB - OBJECTIVE: Our purpose was to evaluate the usefulness of prenatal ultrasonography among women with a positive screen for fetal Down syndrome on the basis of three biochemical markers--maternal serum alpha-fetoprotein, human chorionic gonadotropin, and unconjugated estriol. STUDY DESIGN: A total of 395 women underwent prenatal ultrasonography at a single institution after being identified as screen positive (midtrimester risk > or = 1:195) on the basis of triple-marker screening between 15 and 18 weeks. Ultrasonographic findings were compared with the biochemical markers and the eventual fetal outcome for these patients. Ultrasonographic abnormalities that were evaluated included structural defects, nuchal thickening or cystic hygroma, echogenic bowel, cerebral ventricular dilatation, pyelectasis, and shortened femur. RESULTS: Among 395 patients, 374 (94.7%) had normal karyotype by genetic amniocentesis (n = 232) or postnatal follow-up (n = 142), 18 (4.5%) proved to have Down syndrome, and three had other karyotypic abnormalities. One or more ultrasonographic abnormalities were found in nine of 18 (50%) with Down syndrome compared to 27 of 377 (7.2%) other fetuses (p < 0.001). Fetuses with abnormal ultrasonography results included three with other chromosome abnormalities and five with nonchromosomal anomalies. An abnormal ultrasonography result increased the risk of Down syndrome by 5.6-fold (25% from 4.5%) and a negative result reduced the risk by 45% (2.5% from 4.5%). The value of ultrasonography is further enhanced when all chromosome abnormalities and nonchromosomal anomalies are considered. CONCLUSION: Abnormal ultrasonographic findings increase the risk for Down syndrome, whereas normal findings are less predictive of normalcy. After correction for inaccurate menstrual dates, genetic amniocentesis should be offered in spite of a normal ultrasonography result among women with positive triple screen. PMID- 7485290 TI - Prevention of diabetes-associated embryopathy by overexpression of the free radical scavenger copper zinc superoxide dismutase in transgenic mouse embryos. AB - OBJECTIVES: It has recently been suggested that oxygen free radicals are involved in the high incidence of fetal dysmorphogenesis that is associated with diabetic pregnancies. The purpose of the current investigation was to study the effect of copper zinc superoxide dismutase, a free radical scavenging enzyme, on the prevention of diabetes-associated embryopathy in mice. STUDY DESIGN: Mice used in this study were either transgenic, bearing the human copper zinc superoxide dismutase gene, or nontransgenic controls. Diabetes was generated by streptozotocin administration on days 6 and 7 of gestation. Hyperglycemia developed on day 8 and was maintained through day 10 (critical period of organogenesis). On day 10 fetuses were examined for external anomalies, and their crown-rump lengths and deoxyribonucleic acid content were determined. RESULTS: Induction of maternal diabetes produced a significant reduction in mean crown rump length of control embryos (4.48 +/- 0.7 mm vs 3.65 +/- 0.6 mm, p = 0.0001), whereas transgenic embryos were not affected (4.72 +/- 0.6 mm vs 4.45 +/- 0.8 mm, p > 0.05). After induction of diabetes fetal loss and malformation rates were significantly higher in control embryos (6.0% vs 23.8% and 8.4% vs 16.5%, respectively). Transgenic embryos were practically unaffected by diabetes and showed fetal loss and malformation rates of 5.9% and 4.4%, respectively, after induction of diabetes. CONCLUSIONS: Elevated levels of copper zinc superoxide dismutase, a key enzyme in the metabolism of free oxygen radicals, elicit a protective effect against diabetes-associated embryopathy. PMID- 7485291 TI - Underlying disorders associated with severe early-onset preeclampsia. AB - OBJECTIVE: Our purpose was to determine whether patients with severe early-onset preeclampsia have hemostatic or metabolic abnormalities that are associated with a tendency to vascular thrombosis. STUDY DESIGN: A total of 101 patients with a history of severe early-onset preeclampsia were tested at least 10 weeks post partum for the presence of hyperhomocysteinemia (methionine loading test), protein C, protein S, and antithrombin III deficiency, activated protein C resistance, lupus anticoagulant, and immunoglobulin G and/or M anticardiolipin antibodies. RESULTS: Of the 101 patients, 39 (38.6%) had chronic hypertension. Of the 85 patients tested for coagulation disturbances, 21 (24.7%) had protein S deficiency. Of the 50 patients tested for activated protein C resistance, 8 (16.0%) were positive. Of the 79 patients tested for hyperhomocysteinemia, 14 (17.7%) had a positive methionine loading test. Finally, 95 patients were tested for anticardiolipin antibodies; 27 (29.4%) had detectable immunoglobulin G and/or M anticardiolipin antibodies. CONCLUSION: Patients with a history of severe early onset preeclampsia should be screened for protein S deficiency, activated protein C resistance, hyperhomocysteinemia, and anticardiolipin antibodies, since these results may have an impact on counseling for and pharmacologic management in future pregnancies. PMID- 7485292 TI - Intrauterine growth restriction in infants of less than thirty-two weeks' gestation: associated placental pathologic features. AB - OBJECTIVE: Our purpose was to describe placental lesions associated with normal and abnormal fetal growth in infants delivered for obstetric indications at < 32 weeks' gestation. STUDY DESIGN: Maternal and neonatal charts and placental tissues from 420 consecutive nonanomalous live-born singleton infants delivered at < 32 weeks' gestation with accurate gestational dates were retrospectively studied. Excluded were cases with maternal diabetes, chronic hypertension, hydrops fetalis, diagnosed congenital viral infection, and placenta previa, leaving four primary indications for delivery: preeclampsia, preterm labor, premature rupture of membranes, and nonhypertensive abruptio placentae. The presence and severity of placental lesions was scored by a pathologist blinded to clinical data. Birth weight and length percentiles were calculated from published nomograms. Asymmetric intrauterine growth retardation (n = 32) was defined as birth weight < 10th percentile with length > 10th percentile and symmetric intrauterine growth retardation (n = 48) as both weight and length < 10th percentile for gestational age. A "growth restriction index" was developed to express a continuum of growth in both length and weight. Contingency tables, analyses of variance, and multiple regression analysis defined significance as p < 0.05 (with corrections for multiple comparisons). RESULTS: A greater proportion of cases with intrauterine growth retardation had lesions of uteroplacental insufficiency (p < 0.001) or chronic villitis (p < 0.02) than did appropriately grown preterm infants. Cases with asymmetric intrauterine growth retardation tended to have more lesions than did cases with appropriate-for-gestational-age infants. Four multiple regression analyses used the growth restriction index as outcome and the histologic lesion that had significant relationships to fetal growth as independent predictors in univariate analyses. Overall, uteroplacental fibrinoid necrosis, circulating nucleated erythrocytes, avascular terminal villi, and villous infarct were significant independent predictors of fetal growth (adjusted R2 = 0.312). With addition of preeclampsia as a variable, villous fibrosis, avascular villi, infarct, and preeclampsia were independent predictors of fetal growth (adjusted R2 = 0.341). In the 65 preeclampsia cases no histologic lesion was an independent predictor of fetal growth, whereas in the nonpreeclampsia cases, villous fibrosis and avascular villi were independent predictors of fetal growth (adjusted R2 = 0.075). CONCLUSIONS: In nonanomalous preterm infants intrauterine growth retardation is most commonly symmetric and is primarily related to the cumulative number and severity of lesions reflecting abnormal uteroplacental or fetoplacental blood flow. The growth restriction index may contribute to the study of the biologic range of fetal growth. The statistical relationship of most placental lesions to intrauterine growth retardation depends on the presence or absence of preeclampsia. PMID- 7485293 TI - Relationship between placental histologic features and umbilical cord blood gases in preterm gestations. AB - OBJECTIVE: Our purpose was to test the hypothesis that placental histologic lesions reflect abnormal placental respiratory function in preterm gestations. STUDY DESIGN: A retrospective study of preterm deliveries from 22 to 32 weeks revealed 431 patients with umbilical venous or arterial blood gas values. Excluded were stillbirth, multiple gestations, placenta previa, maternal medical diseases, and fetal anomalies. Charts were reviewed for principal indication of delivery, diagnosis of labor, and mode of delivery. Blood gases were studied within 10 minutes of delivery on a model 178 automatic pH analyzer (Corning Med, Boston). Placental data included uteroplacental vascular lesions and related villous lesions, lesions of acute inflammation, chronic inflammation, and coagulation. Contingency tables and analysis of variance considered p < 0.05 as significant. RESULTS: Mean +/- SD umbilical vein pH was 7.36 +/- 0.07 (range 6.94 to 7.56) and umbilical artery pH was 7.30 +/- 0.08 (range 6.83 to 7.55). Increasing severity of uteroplacental thrombosis, villous lesions reflective of uteroplacental vascular pathologic mechanisms, avascular villi, histologic evidence of abruptio placentae, chronic villitis, and increased circulating erythrocytes were associated with decrease in umbilical vein and artery pH, increase in umbilical vein and artery PCO2, and decrease in umbilical vein and artery PO2. Histologic evidence of acute infection and villous edema were associated with a higher pH and PO2 and a lower PCO2 in both umbilical vein and artery. Umbilical vein or artery base excess was not related to placental lesions. Labor was not related to blood gas values in this data set, although a subset of cases of extremely preterm premature rupture of membranes and preterm labor who labored and were delivered by cesarean section had significantly poorer umbilical venous and fetal arterial blood gas values (all p < 0.005). Lesions related to poorer blood gas values were significantly more frequent in preterm preeclampsia and nonhypertensive abruptio placentae than in premature rupture of membranes or preterm labor. CONCLUSIONS: Changes in umbilical vein and artery pH, PO2, and PCO2 are significantly related to lesions of uteroplacental vascular pathologic mechanisms and intraplacental thrombosis. Placental lesions may be associated with chronic fetal distress by altering fetal oxygen availability and acid-base status. Placental immaturity resulting from prematurity may be associated with inefficient placental respiratory function and an increased likelihood of cesarean delivery in cases of premature rupture of membranes or preterm labor. Altered fetal acid-base balance plus excess numbers of circulating nucleated erythrocytes suggests that placental respiratory function is functionally abnormal when these lesions are present and leads to fetal tissue hypoxia. PMID- 7485294 TI - Histologic evidence of old intrauterine bleeding is more frequent in prematurity. AB - OBJECTIVE: Our purpose was to study the incidence and location of histologic evidence of intrauterine bleeding in preterm and term placentas. STUDY DESIGN: A total of 462 consecutive placentas delivered at < 32 weeks' gestation, from which cases of placenta previa, stillbirth, and multiple gestation were excluded, were compared with 108 consecutive term placentas (with similar exclusion criteria) in regard to the presence of hemosiderin in decidua of basal plate or placental membranes. Of the 462 preterm cases, 448 charts made specific reference to the presence or absence of vaginal bleeding. Bloody show alone was not considered bleeding. The blinded scoring of lesions (including acute ascending infection, uteroplacental vascular pathologic processes and related ischemic damage, chronic inflammation, and coagulation related lesions) was analyzed by contingency tables (p < 0.05 significant). RESULTS: A total of 196 of 462 (43%) preterm placentas had any decidual hemosiderin compared with one of 108 (0.8%) at term (p < 0.00001). Among the preterm cases, hemosiderin was significantly more common in preeclampsia (45/76, 60%) and in cases clinically diagnosed as nonhypertensive abruptio placentae (21/33, 64%) than in premature membrane rupture (72/192, 37.5%) and preterm labor (58/161, 36%, p < 0.003). The incidence of placental lesions in preterm cases with extraplacental membrane hemosiderin was not different than it was in cases without hemosiderin. Placental lesions related to basal-plate decidual hemosiderin in the preterm cases were villous infarct (p < 0.0001), uteroplacental vessels with absence of physiologic change (p < 0.003) and increased numbers of circulating nucleated erythrocytes (p < 0.0007), uteroplacental thrombosis (p < 0.0001), and villous fibrosis (p < 0.0001) and hypovascularity (p < 0.0001). Among the preterm cases, 23 of 48 (48%) with first trimester bleeding, 33 of 66 (50%) with second-trimester bleeding, and 31 of 64 (48%) with multiple episodes of bleeding had decidual hemosiderin (p < 0.0001). A clinical history of gestational bleeding was significantly less common in cases of preterm preeclampsia with histologic evidence of bleeding (four of 73, 5.5%) than in nonhypertensive abruptio placentae (18/31, 58%), premature rupture of membranes (52/183, 28%), or preterm labor (31/161, 19%, p < 0.0001). Hemosiderin was not related to clinical bleeding < 72 hours of delivery (p > 0.20). CONCLUSIONS: Decidual bleeding is common in all clinical types of preterm birth and is most common in preterm preeclampsia and nonhypertensive abruption placentae. A clinical history of bleeding is not correlated with the presence of decidual hemosiderin. Bleeding in the basal plate is related to histologic evidence of chronic uteroplacental vascular pathologic processes, which in cases of spontaneous prematurity (premature rupture of membranes, preterm labor, nonhypertensive abruptio placentae) may be associated with decidual bleeding which occasionally may be clinically manifested as gestational bleeding. PMID- 7485295 TI - Early ultrasonographic detection of fetal growth retardation in an ovine model of placental insufficiency. AB - OBJECTIVE: Our aims were as follows: (1) to determine whether fetal growth retardation can be detected noninvasively with ultrasonography in ewes and (2) to establish the time interval between exposure of ewes to environmental stress that causes growth retardation (heat stress) and detection of growth lag for specific fetal body measurements. STUDY DESIGN: Four ewes were exposed to heat stress for 80 days starting at 35 days' gestation. (The duration of pregnancy in sheep is 147 days). Serial ultrasonographic measurements of fetal biparietal diameter, abdominal circumference, and femur and tibia lengths were obtained beginning at 50 days' gestation. Growth curves were calculated for each parameter and compared with growth curves obtained from 43 normal fetuses. RESULTS: Biparietal diameter measurements deviated significantly from normal starting at 90 days' gestation (p < 0.05). Abdominal circumference diverged at 70 days' gestation (p < 0.05), and both femur and tibia length diverged at 80 days (p < 0.05). The regression lines showed significant differences for all the parameters in both slope (p < 0.01) and intercept (p < 0.01). CONCLUSIONS: Fetal growth retardation can be detected noninvasively by ultrasonography after approximately 5 weeks of exposure to heat stress. Fetal growth continues throughout gestation but at a slower rate than normal and according to a pattern similar to that observed in asymmetrically growth-retarded human fetuses. Early detection of stunted fetal growth in an animal model is important for testing intervention strategies in the treatment of fetal growth retardation. PMID- 7485296 TI - Meconium inhibits the contraction of umbilical vessels induced by the thromboxane A2 analog U46619. AB - OBJECTIVE: Our objective was to study the direct effects of meconium on isolated umbilical artery and vein in vitro. STUDY DESIGN: Intact umbilical artery and vein rings were suspended in 5 ml organ baths containing kreb's solution (pH 7.2, 37 degrees C; bubbled with 2.5% oxygen and 8.07% carbon dioxide, balance nitrogen) for isometric tension recording. Meconium alone (final bath concentration 1%), centrifuged and filtered meconium, and meconium with the lipid fraction removed (separated into < 30,000 kd and > 30,000 kd) were added to the baths. Some vessels were also incubated in 1% meconium for 30 minutes, after which the meconium was washed out. Concentration-response curves to U46619 were obtained. The negative log of the concentration that evokes 50% of the maximal contraction was determined. RESULTS: Umbilical artery and vein had no sustained response to meconium. Tension that developed by rings exposed to meconium was significantly less than control at all concentrations of U46619. There was some loss of the efficacy of meconium after centrifugation-filtration and washing out. Meconium without the lipid fraction had less inhibitory effect than did native meconium. CONCLUSIONS: Meconium does not cause sustained contraction of umbilical artery and vein in vitro. It does, however, inhibit the contractile effect of U46619 in both umbilical artery and vein smooth muscle. The inhibitory activity of meconium appears to be concentrated in the lipid fraction. PMID- 7485298 TI - The efficacy of the placental biopsy. AB - OBJECTIVE: Our purpose was to determine the sensitivity and specificity of pathologic diagnoses made from a placental biopsy specimen compared with diagnoses made from a complete placental examination. STUDY DESIGN: Biopsy was performed on 200 singleton placentas with a 16-gauge Rutner biopsy needle shortly after delivery. The biopsy specimens and placentas were evaluated by standard placental pathologic criteria. RESULTS: The presence of villous edema on the biopsy specimen led to the diagnosis of placental villous edema with a sensitivity of 51% and specificity of 86%, yielding a positive predictive value of 0.97. The sensitivity of the biopsy diagnosis of "increased syncytial knots" was 86%, whereas the specificity was 82%, yielding a positive predictive value of 0.90. CONCLUSIONS: Because a placental biopsy specimen after delivery is reasonably sensitive for diagnosing villous abnormalities that reflect acute and chronic stresses to the placenta, it may be useful to develop a placental biopsy that can be performed safely during pregnancy. Such a biopsy could be the basis for the rational treatment of some diseases of pregnancy. PMID- 7485297 TI - Preinduction cervical ripening with commercially available prostaglandin E2 gel: a randomized, double-blind comparison with a hospital-compounded preparation. AB - OBJECTIVE: Our purpose was to compare the efficacy, safety, and cost of commercially available dinoprostone cervical gel 0.5 mg with a hospital compounded formulation. STUDY DESIGN: One hundred thirty-four patients undergoing labor induction were randomly assigned to one of two cervical ripening groups. Commercially available dinoprostone gel 0.5 mg or a compounded formulation of 0.5 mg of prostaglandin E2 gel was administered endocervically. On the basis of cervical scores, gel was reapplied at 6-hour intervals for a maximum of three doses. Physicians managing labor were blinded as to treatment group allocation. RESULTS: Among 134 patients evaluated, 70 were allocated to the commercially available gel and 64 to the compounded gel. No statistically significant differences were noted between the treatment groups with respect to start-to delivery interval, number of doses, amount of oxytocin, or neonatal adverse outcomes. However, cesarean delivery was performed less frequently in patients in the group receiving the commercially available gel (12.9%) than in the group receiving the compounded gel (28.1%) (p = 0.03). Because of the higher cesarean delivery rate in the compound group, use of this formulation was not associated with cost savings. CONCLUSIONS: The two prostaglandin E2 formulations appeared equivalent with respect to efficacy. An unexplained higher cesarean section rate, however, was associated with the use of the compounded preparation. PMID- 7485300 TI - Placental pathologic features of preterm preeclampsia. AB - OBJECTIVE: Our purpose was to compare the incidence and interrelationships of uteroplacental vasculopathy and chronic inflammatory and placental vasoocclusive lesions in preeclampsia and spontaneous delivery before 32 weeks' gestation. STUDY DESIGN: Review of singleton live-born nonanomalous infants born at 22 to 32 weeks' gestation identified 76 cases of preeclampsia and 353 cases of spontaneous prematurity (spontaneous premature membrane rupture [n = 192], preterm labor, intact membranes [n = 161]). Histologic lesions were considered as belonging to one of five major pathophysiologic groups: (1) uteroplacental vascular lesions and related villous lesions, (2) chronic inflammatory lesions, (3) coagulation related lesions, (4) acute inflammatory lesions, and (5) unclassified lesions. Contingency table analyses considered p < 0.05 significant. Factor analysis extracted combinations of related variables. RESULTS: More frequent in preeclampsia versus spontaneous prematurity were chronic uteroplacental vasculitis (29% vs 20%, p < 0.05), chronic villitis (20% vs 3%, p < 0.001), avascular villi (39% vs 16%, p < 0.001), and "hemorrhagic endovasculitis" (9% vs 2.5%, p < 0.03). In the preeclampsia cases factor analysis extracted 13 categories of related lesions. Four categories contained uteroplacental vascular lesions. Five categories included lesions related to chronic inflammation, and eight included lesions related to coagulation. Four categories loaded lesions from one major pathophysiologic group only. Three categories loaded lesions from all three pathophysiologic groups. Unclassified lesions loaded into two factor categories that were unrelated to the other lesions. CONCLUSIONS: Chronic inflammatory and placental vasoocclusive lesions are more common in preterm preeclampsia than in spontaneous prematurity. Immunopathologic processes and coagulation may be involved in the pathophysiologic mechanisms of preterm preeclampsia independent of uteroplacental vascular pathologic features. PMID- 7485299 TI - Insulin and glucose modulate glucose transporter messenger ribonucleic acid expression and glucose uptake in trophoblasts isolated from first-trimester chorionic villi. AB - OBJECTIVE: Our purpose was to determine the effects of insulin and glucose on glucose transport and expression of GLUT1 glucose transporter messenger ribonucleic acid in first-trimester human trophoblast-like cells. STUDY DESIGN: First-trimester human trophoblast-like cells were maintained as a continuous cell line. For 2[3H]deoxy-D-glucose uptake and messenger ribonucleic acid studies the cells were incubated in the presence or absence of insulin (10(-7) to 10(-11) mol/L) or D-glucose (0 to 50 mmol/L) for 0 to 24 hours. Glucose transport was measured by incubating cells with 0.1 mmol/L 2[3H]deoxy-D-glucose for 5 minutes. Specific uptake was determined by incubating companion cultures with 10 mumol/L cytochalasin B. The cells were then solubilized with sodium hydroxide and the radioactivity counted. Data were expressed as nanomoles of 2[3H]deoxy-D-glucose transported per milligram of protein per 5 minutes and analyzed by one-way analysis of variance with post hoc testing by the method of Tukey. GLUT1 messenger ribonucleic acid was measured by Northern blotting of total ribonucleic acid samples hybridized to a phosphorus 32-labeled complementary deoxyribonucleic encoding the rat GLUT1 glucose transporter. As a control for loading efficiency, blots were stripped and rehybridized to a 40-mer phosphorus 32-labeled beta-actin oligonucleotide probe. RESULTS: Insulin treatment resulted in a dose-dependent increase in the transport of 2[3H]deoxy-D-glucose at 24 hours (p < 0.001 at 10( 7) mol/L). This change was first detected at 12 hours of incubation. These data closely paralleled the insulin-induced increase in GLUT1 messenger ribonucleic acid seen in Northern blots. In contrast to insulin, increasing concentrations of D-glucose did not change the transport of 2[3H]deoxy-D-glucose. However, when cells were incubated in low concentrations of D-glucose (0 or 1 mmol/L), an enhancement in the uptake of 2[3H]deoxy-D-glucose (p < 0.001) was observed. Kinetic studies indicated that D-glucose augmentation of 2[3H]eoxy-D-glucose uptake was significant at 9 hours (p < 0.05). The effects of D-glucose on GLUT1 messenger ribonucleic acid expression paralleled the uptake of 2[3H]deoxy-D glucose, although the modulation of GLUT1 messenger ribonucleic acid levels by glucose was much less pronounced than in insulin-treated cells. CONCLUSION: Although it has been assumed that the placenta has a limited role in influencing glucose transport to the fetus, our in vitro data demonstrate that both insulin and glucose can modulate glucose transport at the cellular level of the placental trophoblast. Thus maternal insulin and glycemic status may influence the expression of GLUT1, the major trophoblast glucose transporter protein, therefore directly affecting first-trimester placental glucose transport. These in vitro data may help explain the association between maternal glucose abnormalities and impaired fetal development during the first trimester when placental GLUT1 messenger ribonucleic acid expression is at its peak. PMID- 7485301 TI - Normal nuchal thickness in the midtrimester indicates reduced risk of Down syndrome in pregnancies with abnormal triple-screen results. AB - OBJECTIVE: Our purpose was to determine whether nuchal thickness measurement can identify the euploid fetuses in midtrimester pregnancies at increased risk for Down syndrome on the basis of maternal age and serum screening. STUDY DESIGN: Nuchal thickness was obtained prospectively in 651 consecutive fetuses at 14 to 21 weeks' gestation and at > or = 1:270 risk for Down syndrome on the basis of unconjugated estriol, alpha-fetoprotein, and human chorionic gonadotropin levels. The risk of Down syndrome with a normal nuchal thickness was determined. A receiver-operator characteristic curve was used to determine a serum-based risk threshold below which the risk for Down syndrome was low. The prevalence of Down syndrome in fetuses with both a normal nuchal thickness and a below-serum-risk threshold was compared with prevalence in either those above threshold risk or with an abnormal nuchal thickness. RESULTS: There were eight cases of trisomy 21 and one case each of 46,XX/47,XXX, 46,XY/47,XY, +7, and 46,XX, 11q-. The sensitivity of an abnormal nuchal thickness (> or = 6 mm) for detecting Down syndrome was four in eight (50%) (95%) confidence interval 15.3% to 84.6%). The risk of Down syndrome was significantly increased with an abnormal compared with a normal nuchal thickness, four in 13 (30.8%) versus four in 638 (0.6%), p < 0.0001. A risk threshold was defined at > or = 1:100 on the basis of the receiver operator characteristic plot. Of 390 cases with a normal nuchal thickness and a serum risk estimate < 1:100, there were no cases of Down syndrome (0/390 vs 8/253, p = 0.002). CONCLUSION: Normal nuchal thickness significantly reduces the risk of Down syndrome and may help reduce the number of amniocenteses done for abnormal triple screen results. PMID- 7485302 TI - A comparison between two screening methods for detection of microproteinuria. AB - OBJECTIVE: We compared two screening tests for microproteinuria with 24-hour quantitative measurements to determine which method is better at predicting clinically significant proteinuria. STUDY DESIGN: We obtained 690 24-hour urine collections from both low- and high-risk patients seen for prenatal care. Qualitative screening for microproteinuria on the basis of the protein-error-of indicators principle (Ames Multistix 10SG and Micro-bumintest, Miles Diagnostic Division, Elkhart, Ind.) was done by the same investigator (C.S.). Quantitative assay was done by use of pyrogallol red-molybdate for total protein and by radioimmunoassay for albumin. RESULTS: The Micro-bumintest had a sensitivity of 87% compared with 36% for the Multistix 10SG. It also had a higher specificity and higher positive and negative predictive values. The Micro-bumintest was a better screening test in patients with significant protein excretion (> 300 mg/24 hours). CONCLUSION: The Micro-bumintest has a much higher sensitivity and a lower false-negative rate than does the Multistix 10SG. Our data support the Micro bumintest as a better screening test for clinically significant proteinuria. PMID- 7485303 TI - Uterine artery Doppler velocimetry in the detection of adverse obstetric outcomes in women with unexplained elevated maternal serum alpha-fetoprotein levels. AB - OBJECTIVE: We hypothesized that in pregnancies complicated by unexplained elevations of maternal serum alpha-fetoprotein, second-trimester uterine artery Doppler findings would detect adverse obstetric outcomes. STUDY DESIGN: One hundred three subjects with unexplained elevations of maternal serum alpha fetoprotein had uterine artery Doppler velocimetry studies performed at the time of targeted ultrasonographic examination (17 to 22 weeks). A resistance index > 95th percentile or the presence of a uterine notch was considered abnormal. Adverse outcomes included preeclampsia, preterm birth, low birth weight, intrauterine growth restriction, abruptio placentae, and fetal death. Statistical analysis was performed by Student t test, chi 2 analysis, and stepwise logistic regression analysis. RESULTS: An elevated uterine resistance index was associated with an increased relative risk for both preeclampsia (relative risk 41.82, 95% confidence interval 5.36 to 326.13) and low birth weight (relative risk 4.65, 95% confidence interval 1.90 to 11.39). A uterine artery notch was associated with an increased risk of preeclampsia (relative risk 52.22, 95% confidence interval 6.82 to 399.70), preterm birth (relative risk 3.21, 95% confidence interval 1.32 to 7.81), and low birth weight (relative risk 4.18, 95% confidence interval 1.64 to 10.66). When the presence of a uterine notch, vaginal bleeding, and level of maternal serum AFP were analyzed by stepwise logistic regression, the presence of a notch was found to be the only significant factor (odds ratio 6.95, 95% confidence interval 1.24 to 39.10) for the development of an adverse outcome. CONCLUSIONS: Abnormal uterine artery Doppler findings are associated with an increased frequency of adverse obstetric outcomes in women with unexplained elevated maternal serum AFP levels. Abnormal Doppler findings demonstrated high sensitivity for the development of preeclampsia but were less sensitive in predicting other outcomes. The presence of a uterine artery notch is a better independent predictor of adverse outcome than are early vaginal bleeding or maternal serum AFP level. PMID- 7485306 TI - Fetal cardiac hypertrophy and cardiac function in diabetic pregnancies. AB - OBJECTIVE: Our purpose was to evaluate the relationship between fetal cardiac wall hypertrophy and ventricular function in fetuses of metabolically controlled, insulin-requiring diabetics. STUDY DESIGN: M-mode directed fetal echocardiography included measurements of left and right ventricular free wall and interventricular septal thickness and ventricular diastolic and systolic dimensions. Fetal measurements included biparietal diameter, estimation of fetal weight, and cardiac area/thoracic area. Cardiac size is expressed as a ratio of wall thickness/biparietal diameter, and function is expressed as ventricular shortening fraction (Ventricular diastolic dimension-Ventricular systolic dimension/Ventricular diastolic dimension). Fetuses of diabetics at 20 to 24, 28 to 31, and 32 to 36 weeks' gestation were compared with normal fetuses of nondiabetic mothers at similar gestational ages, which were used as a control group. RESULTS: Study and control groups were comparable in mean gestational age (22 vs 20, 29 vs 29, 35 vs 34 weeks) in each of groups 1, 2, and 3. All fetuses tested fell within these groups. Estimated fetal weight, cardiac area/thoracic area, right ventricular wall thickness/biparietal diameter, and interventricular septal thickness/biparietal diameter were greater in the study group between 32 and 36 weeks: 3227 +/- 430 versus 2235 +/- 176 gm (p < 0.05), 0.32 versus 0.29, (p < 0.05), 0.53 +/- 0.05 versus 0.44 +/- 0.05 mm (p < 0.05), and 0.50 +/- 0.3 versus 0.46 +/- 0.5 mm (p < 0.05), respectively. Right ventricular shortening fraction/left ventricular shortening fraction was significantly different from controls in this group (1.30 vs 0.89, p < 0.05). This change was a function of altered right ventricular shortening fraction in late diabetic pregnancy. Right ventricular shortening fraction in the study groups was 0.39, 0.36, and 0.52, respectively, versus 0.36, 0.41, and 0.33 in controls. Left ventricular shortening fraction in the group at 32 to 36 weeks and in controls in three groups was 0.42, 0.39, 0.40 and 0.44, 0.37. CONCLUSION: These findings reveal an increase in right ventricular shortening fraction associated with global cardiac enlargement. Myocardial hypertrophy involving right ventricular wall thickness and interventricular septal thickness in metabolically stable insulin-requiring diabetics revealed hypercontractility of the right ventricle. PMID- 7485305 TI - Neural network prediction of nonstress test results: how often should we perform nonstress tests? AB - OBJECTIVE: Our purpose was to predict outcomes and optimal intervals for nonstress tests of term gravid women with neural networks. STUDY DESIGN: We studied 100 normal term patients whose 30-minute nonstress tests, performed on 5 consecutive days, were computer analyzed for the following elements: fetal heart rate baseline, variability, signal loss, accelerations (> 15 beats/min), and decelerations. The training set used 65 patients; the testing, 35 patients. Nonstress test data (days 1 to 4) were inputs; day 5 data were training patterns. Networks for each nonstress test element used Brainmaker Macintosh 1.0 (California Scientific Software, Nevada City, Calif.) trained to 0.12 tolerance. Actual fetal heart rate elements and their daily differences were compared with predictions by the networks and multiple regressions. RESULTS: There was little difference between networks using daily or alternate-day inputs for predicting test performance on day 5; networks using test intervals > 2 days could not be trained to tolerance. Long-term fetal heart rate variation was the nonstress test element best predicted. Daily differences networks provided better prediction of all day 5 data than did actual daily values networks or multiple regression formulas. CONCLUSIONS: Baseline long-term fetal heart rate variability seems to be the most predictable fetal heart rate element over time and should merit more consideration in overall fetal testing. Fetal heart rate elements are not easily predicted by any method for intervals longer than 2 days. Using longer test intervals might run a greater risk for unanticipated changes in nonstress test outcomes, even when fetal condition is normal. PMID- 7485304 TI - A multicenter randomized controlled trial of home uterine monitoring: active versus sham device. The Collaborative Home Uterine Monitoring Study (CHUMS) Group. AB - OBJECTIVE: Our purpose was to determine the efficacy of a home uterine activity monitoring system for early detection of preterm labor and reduction of preterm birth. STUDY DESIGN: A randomized, controlled, double-blinded trial was performed in which pregnant women between 24 and 36 weeks' gestation and at high risk for preterm labor or birth were assigned to receive twice daily nursing contact and home uterine activity monitoring with either active (data revealed) or sham (data concealed) devices. Study end points included mean cervical dilatation and its mean change from a previous visit at preterm labor diagnosis, preterm birth rate, and infant outcomes. Analysis of variance or logistic models including terms for site and group-by-site interaction effects were constructed for all variables. RESULTS: Of 1355 patients enrolled, 1292 were randomized, 1165 used home uterine activity monitoring devices, and 842 (72.3%) completed the study. Both device groups had similar demographics, enrollment and delivery gestational ages, discontinuation rates, risk factors, birth weights, cervical dilatation at enrollment and at preterm labor diagnosis, change in cervical dilatation at preterm labor diagnosis, rates of preterm labor and birth, and neonatal intensive care requirements. Power to detect a difference in cervical dilatation > or = 1 cm at diagnosis of preterm labor was 0.99 for all risk factors. CONCLUSIONS: Uterine activity data obtained from home uterine activity monitoring, when added to daily nursing contact, were not linked to earlier diagnosis of preterm labor or lower rates of preterm birth or neonatal morbidity in pregnancies at high risk for preterm labor and birth. PMID- 7485307 TI - Labor induction with intravaginal misoprostol versus intracervical prostaglandin E2 gel (Prepidil gel): randomized comparison. AB - OBJECTIVE: Our purpose was to compare the safety and efficacy of intravaginal prostaglandin E1, misoprostol, with that of intracervical prostaglandin E2 (Prepidil gel) for labor induction. STUDY DESIGN: One hundred three patients with an indication for induction of labor were randomly assigned to induction with prostaglandin E1, 50 micrograms intravaginally, or with Prepidil gel, 0.5 mg intracervically, every 4 hours until active labor. RESULTS: Four patients were excluded, leaving 49 patients who received prostaglandin E1 and 50 who received prostaglandin E2. The time from start of induction to vaginal delivery was significantly shorter in the prostaglandin E1 group (11.4 vs 18.9 hours, p < 0.001), and fewer patients in the prostaglandin E1 group required oxytocin augmentation (23% vs 55%, p < 0.005). No significant differences were noted in mode of delivery or in adverse maternal, fetal, or neonatal effects. CONCLUSION: Intravaginal prostaglandin E1 is a more effective, lower-cost agent for induction of labor than is intracervical prostaglandin E2 gel and is comparable in safety. PMID- 7485308 TI - The effects of atrial natriuretic factor and angiotensin II on fetal-placental perfusion pressure in the ex vivo cotyledon model. AB - OBJECTIVE: Our purpose was to investigate pressure changes induced by angiotensin II on placental vasculature pretreated with atrial natriuretic factor. STUDY DESIGN: A dual-perfused cotyledon model was used. Two cotyledons from each placenta were perfused. One cotyledon was infused with atrial natriuretic factor for 30 minutes while the other received an equal volume of saline solution. Three atrial natriuretic factor concentrations were studied: 50 pg/ml, 150 pg/ml, and 15 ng/ml. Both cotyledons received injections of angiotensin II, at the following doses: 1 x 10(-11.5), 1 x 10(-11), 1 x 10(-10.5), and 1 x 10(-10) mol. RESULTS: Cotyledons subjected to 50 pg/ml and 150 pg/ml concentrations of atrial natriuretic factor did not differ in pressure responses to angiotensin II, compared with the saline-infused cotyledons (p > 0.05). The 15 ng/ml concentration of atrial natriuretic factor, however, decreased the pressor response of angiotensin II (p < 0.034). CONCLUSIONS: Atrial natriuretic factor decreases vasoconstriction caused by angiotensin II. However, this was seen only at a supraphysiologic concentration. No effect was noted at normal fetal concentrations of atrial natriuretic factor. PMID- 7485309 TI - Quantitative echocardiographic assessment of left ventricular function in critically ill obstetric patients: a comparative study. AB - OBJECTIVE: Our purpose was to compare different echocardiographic methods to quantitate maternal ventricular function. STUDY DESIGN: Eleven critically ill obstetric patients requiring invasive monitoring and M-mode and two-dimensional Doppler echocardiographic studies were retrospectively studied. Ventricular volumes and ejection fraction derived from multiple methods were computed and compared with analysis of variance and Newman-Keuls tests. RESULTS: Two M-mode studies could not be analyzed. M-mode and two-dimensional estimates of stroke volume were similar to the previously validated Doppler stroke volume values of these 11 patients. End-diastolic volume estimates were similar. There was no difference in ejection fraction for the five different two-dimensional Doppler methods used (66%, 66%, 67%, 63%, and 63%). CONCLUSION: Left ventricular function is reliably calculated in pregnant women by a number of different echocardiographic techniques. The ability to combine echocardiographic techniques is helpful when any single method does not provide all of the information required. PMID- 7485310 TI - Pregnancy outcome for women with very low levels of maternal serum unconjugated estriol on second-trimester screening. AB - OBJECTIVE: Our purpose was to assess pregnancy outcome for women whose second trimester triple screen results revealed very low unconjugated estriol levels. STUDY DESIGN: Of 44,146 women screened for alpha-fetoprotein, human chorionic gonadotropin, and unconjugated estriol, a group of women with isolated very low unconjugated estriol levels were selected as the study group. Pregnancy outcome was obtained by contacting the referring physicians. RESULTS: During the study period 62 women had unconjugated estriol levels at or below the sensitivity of the assay. Fetal death was the most common explanation, accounting for 37.1% of cases. Six cases of early death followed uneventful prenatal visits. Eighteen women had unexplained very low unconjugated estriol levels. Of these 18, four cases of placental sulfatase deficiency were diagnosed. Sixteen of these 18 women gave birth to male infants (p = 0.03). CONCLUSION: A very low unconjugated estriol level in the second trimester is associated with increased risk for early death and placental sulfatase deficiency. PMID- 7485311 TI - Fetal liver cell transplantation for the creation of lymphohematopoietic chimerism in fetal baboons. AB - OBJECTIVE: Our purpose was to create xenogeneic lymphohematopoietic chimerism by in utero transplantation of human fetal liver cells in the midgestation fetal baboon. STUDY DESIGN: Human fetal liver cell suspensions obtained from preimmune human fetuses (< 80 days' gestation) were injected into the peritoneal cavity of three fetal baboons (85, 95, and 104 days' gestation). A total of 9 x 10(6) cells, in a volume of 1 ml, were injected percutaneously into the fetal abdominal cavity under ultrasonographic guidance. The success of the injection was assessed by observing ascites and free loops of fetal bowel after injection. Fetal umbilical cord blood (35 days posttransplantation) and neonatal blood and bone marrow were obtained to be assayed for the presence of donor hematopoietic cells. Chimerism was detected by fluorescence in situ hybridization with a human Y chromosome specific probe. RESULTS: All the animals survived the in utero procedures. Thirty-five days after transplantation engraftment was noted in one animal. Postnatally the same animal showed engraftment in both the peripheral blood and bone marrow. The rate of chimerism was 1.5% (1.5% of the cells were human) in both the peripheral blood and bone marrow. CONCLUSIONS: This study demonstrates that creation of xenogeneic lymphohematopoietic chimerism is possible in the midgestation fetal baboon. However, the level of chimerism was too low to study the biologic activity of the transplanted cells or to potentially ameliorate lymphohematopoietic disorders. Future studies using allogeneic tissue, evaluating cells obtained from both fetal and adult donors, and comparisons between purified stem cells and fetal liver cells are needed. PMID- 7485312 TI - Differential regulation of seizure activity in the hippocampus of male and female rats. AB - OBJECTIVE: The aim of the current study was to directly examine and compare the susceptibility to N-methyl-D-aspartate-induced seizures in male versus female rats. We also sought to compare the anticonvulsant effects of magnesium sulfate in these two groups. STUDY DESIGN: Eighteen female and 10 male rats were stereotaxically implanted with a chronic bipolar recording electrode in the hippocampus and an injection cannula in the lateral cerebral ventricle. After 1 week rats randomly received an intravenous injection of 90 mg/kg magnesium sulfate or saline solution control. Fifteen minutes after the infusion rats were given the convulsant N-methyl-D-aspartate at a dose of 5 micrograms/microliters by direct intraventricular injection. Electrical seizure activity was thereafter assessed for 20 minutes. All data were analyzed by the Mann-Whitney U test and Student t test. RESULTS: In saline solution-treated rats receiving the convulsant N-methyl-D-aspartate, females had significantly lower total duration (p < 0.01) and total number of seizures (p < 0.05) compared with the male rats. The initial seizure was not affected by gender. In the female animals magnesium sulfate significantly reduced first seizure duration (p < 0.01) compared with saline solution controls. In males magnesium sulfate reduced both total duration (p < 0.05) and total seizure number (p < 0.05) compared with saline solution-treated animals. CONCLUSION: N-methyl-D-aspartate-induced seizure activity is more severe in males versus female rats. Magnesium sulfate's effect on N-methyl-D-aspartate induced seizures is also dependent on gender. We speculate that seizure regulation may be hormonally influenced. PMID- 7485313 TI - Effects of intracervical prostaglandin E2 on fetal heart rate and uterine activity patterns in the presence of oligohydramnios. AB - OBJECTIVE: Our purpose was to compare fetal heart rate patterns and uterine activity before and after preinduction prostaglandin E2 administration in the presence or absence of oligohydramnios. STUDY DESIGN: In a retrospective case controlled review we examined cases in which prostaglandin E2 (Prepidil) was inserted intracervically for gravid women requiring an induction of labor in the presence of either oligohydramnios (amniotic fluid index < or = 5.0) or adequate fluid (amniotic fluid index 5.1 to 23.9). Uterine activity and fetal heart rate tracings that were begun 1 hour before and continued for 6 hours after dosing were interpreted without knowledge of amniotic fluid volume. RESULTS: Cases in the oligohydramnios (n = 51) and adequate fluid (n = 49) groups were the same for maternal age, race, parity, gestational age, and predose Bishop score. Patients with oligohydramnios had more high-amplitude contractions in the first hour after dosing (9.0 +/- 1.2 vs 6.1 +/- 0.9, p < 0.05), but there were no significant differences in the frequency or duration of contractions during the subsequent 5 hours. Uterine hyperstimulation was not seen, and there were no differences in the frequency of variable or late fetal heart rate decelerations. CONCLUSION: For pregnancies undergoing preinduction cervical ripening with intracervical prostaglandin E2, the presence of oligohydramnios was not associated with a greater risk of fetal heart rate decelerations, although contractions were more common during the first hour after dosing. PMID- 7485314 TI - Head entrapment and neonatal outcome by mode of delivery in breech deliveries from twenty-four to twenty-seven weeks of gestation. AB - OBJECTIVES: Our purpose was to determine whether there are differences in the incidence of head entrapment and adverse neonatal outcome by mode of delivery in breech deliveries from 24 to 27 weeks of gestation. STUDY DESIGN: Charts of 132 viable consecutive singleton breech deliveries from 24 to 27 weeks' gestation were reviewed. Statistical methods used included chi 2, logistic regression, and analysis of variance. RESULTS: Head entrapment occurred in four of 43 (9.3%) vaginally delivered neonates and five of 89 (5.6%) neonates delivered by cesarean section (p = 0.17). The only identifiable risk factor for head entrapment was the birth weight category 1000 to 1249 gm (odds ratio 6.4, 95% confidence interval 1.6 to 26.1). There were no statistically significant associations between head entrapment and adverse neonatal outcomes. CONCLUSIONS: There did not appear to be a difference in the incidence of head entrapment by mode of delivery for breech infants at 24 to 27 weeks' gestation, nor did there appear to be a difference in adverse neonatal outcomes after entrapment. However, this latter conclusion has limited power because of the small number of head entrapment cases. PMID- 7485315 TI - Anthropometric estimation of neonatal body composition. AB - OBJECTIVE: Estimation of neonatal body composition can be useful in the understanding of fetal growth. However, body composition methods such as total body water and total body electric conductivity are expensive and not readily available. Our primary purpose was to develop an anthropometric model to estimate neonatal body composition and prospectively validate the model against total body electric conductivity and secondarily to compare our anthropometric model and a previously published anthropometric formula with total body electric conductivity. STUDY DESIGN: A total of 194 neonates had estimates of body composition according to total body electric conductivity (group 1). Parental morphometrics, gestational age, race, sex, parity, and neonatal measurements including birth weight, length, head circumference, and skinfolds (triceps, subscapular, flank, and thigh) were correlated with body fat by use of stepwise regression analysis. The model was validated in a second group of 65 neonates (group 2). RESULTS: There were no significant differences in any of the parental or neonatal measurements between groups 1 and 2. In group 1, 78% of the variance in body fat with the use of total body electric conductivity was explained by birth weight, length, and flank skinfold (R2 = 0.78, p = 0.0001). When prospectively validated by the subjects in group 2, the model had significant and stronger correlation (R2 = 0.84, p = 0.0001) with body fat estimated by total body electric conductivity as compared with the other anthropometric model (R2 = 0.54, p = 0.0001). There was no significant (p = 0.11) difference between our anthropometric estimate of body fat and total body electric conductivity. CONCLUSIONS: The anthropometric model developed can be used to reasonably predict neonatal body fat mass at birth. PMID- 7485316 TI - Accuracy of prenatal determination of RhD type status by polymerase chain reaction with amniotic cells. AB - OBJECTIVE: Our purpose was to determine the accuracy of RhD typing by use of amniocytes obtained at amniocentesis in RhD-negative women. STUDY DESIGN: One hundred thirty-five RhD-negative women undergoing amniocentesis for management of suspected alloimmunization (n = 95) or routine second-trimester cytogenetic indications (n = 40) were studied. Amniocytes were then used as template to amplify specific portions of the Rh D and Rh CcEe genes by polymerase chain reaction. The fetal RhD type was confirmed by serologic techniques either after fetal blood sampling or cord blood samples at birth. RESULTS: Thirty-six fetuses were serologically typed as RhD negative and all 36 were typed RhD negative by polymerase chain reaction. Ninety-eight fetuses were serologically typed as RhD positive; of these, 96 were correctly typed as RhD positive and two were incorrectly typed as RhD negative, with an overall error rate of 1.4%. Both of the errors occurred in a single batch of six samples tested at the same time. In one of these cases the fetus had mild Rh alloimmune disease and required exchange transfusion at birth. In the second case the fetus had severe hydrops fetalis and died in utero at 28 weeks. Deoxyribonucleic acid isolated from fetal blood was tested with the same polymerase chain reaction technique after delivery, and in both cases the fetus was correctly typed as RhD positive. Deoxyribonucleic acid amplification repeatedly failed in one case. CONCLUSION: Prenatal fetal RhD typing by polymerase chain reaction with amniotic fluid cells is accurate and reliable. In sensitized pregnancies it allows early management of Rh disease and avoids invasive procedures in RhD-negative fetuses. In nonsensitized pregnancies it avoids the use of anti-RhD immunoglobulin after invasive procedures or during pregnancy. To eliminate the possibility of genetic and laboratory sources of errors, we suggest using different sets of primers at two different loci (e.g., exon 4 to 5 and exon 10). PMID- 7485317 TI - Acute maternal hydration in third-trimester oligohydramnios: effects on amniotic fluid volume, uteroplacental perfusion, and fetal blood flow and urine output. AB - OBJECTIVE: Our purpose was to determine whether acute maternal hydration in pregnancies with third-trimester oligohydramnios (1) increases amniotic fluid index and hourly fetal urine production rate and (2) alters uteroplacental perfusion and fetal blood flow. STUDY DESIGN: Ten women with third-trimester oligohydramnios (amniotic fluid index < or = 5 cm) and 10 controls with normal amniotic fluid volume (amniotic fluid index > 7 cm) were prospectively recruited for this study. Maternal plasma and urine osmolality, amniotic fluid index, hourly fetal urine production rate, and Doppler flow velocimetry of maternal uterine artery and fetal umbilical, descending aorta, middle cerebral, and renal arteries were determined before and after oral hydration by having the patient drink 2 L of water over 2 hours. RESULTS: There was a significant reduction in maternal plasma (p < 0.05) and urine osmolality (p < 0.0001) in both groups after short-term oral hydration. Hydration increased amniotic fluid volume in women with oligohydramnios (mean change in amniotic fluid index 3.2 cm, 95% confidence intervals 1.1 to 5.3; p < 0.02) but not in those with normal amniotic fluid volume (mean change in amniotic fluid index -2.0, 95% confidence intervals -4.1 to +0.2). The hourly fetal urine production rate, however, did not increase in either group (mean change in hourly fetal urine production rate 3.5 ml/hr, 95% confidence intervals -11.7 to +18.7 and -6.8 ml/hr, 95% confidence intervals -2.9 to -10.7, respectively). Hydration was associated with an increase in uterine artery mean velocity in the oligohydramnios group (mean change in mean velocity 16.7 cm/sec, 95% confidence intervals 8.0 to 25.3; p < 0.006) but not in controls (mean change in mean velocity 1.2 cm/sec, 95% confidence intervals -19.7 to +22.1). There was no change in pulsatility index or in velocity in any of the fetal vessels studied in either group. CONCLUSIONS: Short-term maternal oral hydration increases the amniotic fluid index in women with third-trimester oligohydramnios. Although the mechanism for this effect remains unclear, it could not be accounted for by fetal urination in this study but instead was associated with improved uteroplacental perfusion. PMID- 7485318 TI - A new method for on-screen ultrasonographic determination of fetal cardiac axis. AB - OBJECTIVE: Our purpose was to develop an ultrasonographic method of using multiple electronic calipers for on-screen measurement of fetal cardiac axis. STUDY DESIGN: Two hundred low-risk patients who were seen for antenatal ultrasonography were studied. Standard biometry, anatomic survey, and echocardiography were performed on all fetuses. By use of a four-chamber view fetal cardiac axis was measured with electronic calipers. A table, derived from a trigonometric formula, was created to convert the caliper measurements to cardiac axis in degrees. The results were compared with a protractor-determined fetal cardiac axis. RESULTS: All echocardiograms had normal results. Fetal cardiac axis (+/- 2 SD) by the ultrasonographic method was 43 degrees (+/- 16 degrees) versus 43 degrees (+/- 14 degrees) by the protractor method. This difference was not significant. CONCLUSION: An on-screen method to determine fetal cardiac axis by use of multiple calipers is described. It is comparable to a protractor-measured fetal cardiac axis. PMID- 7485319 TI - Gender-specific second-trimester biometry. AB - OBJECTIVE: This study was designed to evaluate the significance of gender differences for commonly used biometric parameters obtained ultrasonographically from second-trimester euploid fetuses. STUDY DESIGN: Gender-specific linear regression equations were developed for gestational age dependent and independent biometry from second-trimester ultrasonographically normal singleton euploid fetuses by use of biparietal diameters and femur and humerus lengths. Regression lines for male fetuses were compared with those for females by determining overlap of 95% confidence intervals at specific points. RESULTS: A total of 288 male fetuses (mean gestational age 16.7 +/- 1.0 weeks) and 251 female fetuses (mean gestational age 16.8 +/- 1.2 weeks) (not significant) were evaluated. Small statistically significant gender differences in fetal biometry were identified. The largest gender difference for gestational age-dependent parameters was a biparietal diameter difference of 1.15 mm at 21 weeks. The largest gender difference for gestational age-independent parameters was a femur length difference of 1.7 mm at a biparietal diameter of 50 mm. CONCLUSION: Statistically, but not clinically, significant gender differences of small magnitude exist for second-trimester fetal biometry, suggesting that gender specific nomograms may be of limited value. PMID- 7485321 TI - Daily antenatal testing in women with severe preeclampsia. AB - OBJECTIVE: Our purpose was to determine whether daily antenatal testing in the expectant management of severe preeclampsia remote from term prevents stillbirth or neonatal compromise at birth. STUDY DESIGN: We reviewed the medical records of 68 women with severe preeclampsia remote from term who underwent expectant management with daily fetal testing until delivery. On admission each patient had reassuring nonstress testing (absence of persistent severe variable or late decelerations), biophysical profile (> or = 6), and amniotic fluid volume (> or = 2 cm maximal vertical pocket before 32 weeks or amniotic fluid index > or = 5 after 32 weeks). RESULTS: There were no stillbirths. Twenty-one patients (31%) had nonreassuring testing necessitating delivery. Two neonatal deaths occurred as a result of complications of prematurity. There were no statistical differences in the cord arterial pH (p = 0.93) or in the 1- and 5-minute Apgar scores (p = 0.18 and p = 0.88, respectively) between those with normal and abnormal antenatal testing. CONCLUSIONS: Because optimizing neonatal outcome is the only reason to prolong pregnancy in women with severe preeclampsia, confirmation of fetal well being is mandatory. Because neither stillbirths nor fetal compromise at birth occurred in patients undergoing daily antenatal testing, we recommend daily testing in patients with severe preeclampsia managed expectantly. PMID- 7485320 TI - Molecular genotyping of fetal platelet antigens with uncultured amniocytes. AB - OBJECTIVE: Amino acid substitutions in platelet membrane glycoproteins result in alloantigens implicated in neonatal alloimmune thrombocytopenia. We report the use of the reverse dot blot technique to genotype the five major fetal platelet alloantigens from amniotic fluid cells. STUDY DESIGN: We evaluated a patient with Bakb platelet antibodies who had a previous pregnancy complicated by fetal intracranial hemorrhage. The father was heterozygous Baka/Bakb, giving the pregnancy a 50% risk for platelet incompatibility between mother and fetus. Amniotic fluid was obtained at 16 weeks. Deoxyribonuleic acid was extracted from uncultured amniocytes and amplified with polymerase chain reaction. These products were hybridized to filters containing oligonucleotides specific for each of the 10 different platelet antigen alleles. Reactivity was detected with a chromogenic substrate. RESULTS: The reverse dot blot genotyping of uncultured amniocytes revealed the fetus to be Baka/Baka, thus not at risk for neonatal alloimmune thrombocytopenia. CONCLUSION: Precise knowledge of fetal platelet type by amniocentesis could obviate the need for fetal blood sampling and significantly alter prenatal management of neonatal alloimmune thrombocytopenia. PMID- 7485322 TI - Are labor abnormalities more common in shoulder dystocia? AB - OBJECTIVE: Our objective was to determine the association between labor abnormalities and shoulder dystocia. STUDY DESIGN: All consecutive cases of shoulder dystocia from January 1986 to August 1994 were reviewed (n = 276). For purposes of comparison a control group of vaginally delivered patients was randomly selected in a 2:1 ratio (n = 600). Charts were reviewed for demographic information, labor and delivery events, and neonatal outcome. RESULTS: Labor abnormalities were comparable in the shoulder dystocia and control groups, both in the active phase and in the second stage. When patients with diabetes and those with macrosomic infants were analyzed separately, no significant differences in labor abnormalities were identified. The rate of operative vaginal delivery was significantly higher in the shoulder group, and one third of the operative deliveries were midpelvic. In addition, the induction rate was higher in the shoulder group. CONCLUSIONS: Our data suggest that labor abnormalities may not serve as clinical predictors for subsequent development of shoulder dystocia, thus emphasizing the unpredictability of this condition. PMID- 7485323 TI - Fetal macrosomia: does antenatal prediction affect delivery route and birth outcome? AB - OBJECTIVE: Our purpose was to determine whether clinical or ultrasonographic prediction of fetal macrosomia influences subsequent delivery route and birth outcome in a clinical setting where macrosomia is not considered an indication for cesarean delivery. STUDY DESIGN: The hospital records of 504 patients delivered of infants weighing > or = 4200 gm between October 1989 and March 1994 were reviewed. Statistical comparisons were made between patients in whom fetal macrosomia was predicted before delivery (n = 102) and those in whom it was not (n = 402). Cesarean delivery, shoulder dystocia, and birth trauma rates were the variables of interest. RESULTS: Cesarean sections were performed in 52% of the "predicted" group deliveries and in 30% of the "not predicted" group (p < 0.01). The increased cesarean delivery rate in the predicted group appeared to be related to an increased incidence of labor inductions (42.5% vs 26.6%, p = 0.005) and a greater proportion of failed inductions. The proportion of patients delivered by cesarean section without a trial of labor was similar in the predicted and not predicted groups (14.7% vs 10.2%, p = 0.21). There was no significant differences in the incidence of shoulder dystocia or the occurrence of birth trauma. CONCLUSIONS: The antenatal prediction of fetal macrosomia is associated with a marked increase in cesarean deliveries without a significant reduction in the incidence of shoulder dystocia or fetal injury. Ultrasonography and labor induction for patients at risk for fetal macrosomia should be discouraged. PMID- 7485324 TI - Does magnesium sulfate affect the length of labor induction in women with pregnancy-associated hypertension? AB - OBJECTIVE: Our purpose was to determine whether seizure prophylaxis with magnesium sulfate prolongs induction of labor in women with pregnancy-associated hypertension. STUDY DESIGN: Women with a singleton pregnancy in vertex presentation between 32 and 42 weeks' gestation who required induction of labor for either preeclampsia, preeclampsia superimposed on chronic hypertension, or transient hypertension were randomized to receive either magnesium sulfate or phenytoin (Dilantin) for seizure prophylaxis. Women with rupture of the membranes, spontaneous contractions resulting in cervical change, or an initial cervical examination showing > 2 cm dilatation and 50% effacement were excluded. RESULTS: Fifty-four women were randomized to receive either magnesium sulfate (n = 28) or phenytoin (n = 26). The two groups were similar for selected maternal antepartum, intrapartum, and postpartum characteristics that have been associated with a prolongation or failure of labor induction. The two groups were also similar for the interval from the start of induction to (1) the first examination > or = 5 cm cervical dilatation and (2) delivery and in the frequency of women requiring cesarean delivery. CONCLUSION: Compared with phenytoin, magnesium sulfate seizure prophylaxis in women with pregnancy-associated hypertension does not prolong the induction of labor nor does it result in an increase in cesarean deliveries. PMID- 7485325 TI - Amniotic fluid interleukin-10 concentrations increase through pregnancy and are elevated in patients with preterm labor associated with intrauterine infection. AB - OBJECTIVE: To better understand the role of the antiinflammatory cytokine interleukin-10 in preterm labor and infection, we evaluated the amniotic fluid interleukin-10 concentrations through pregnancy, in term, and in preterm labor. STUDY DESIGN: Amniotic fluid interleukin-10 levels were measured in 147 women throughout pregnancy including patients in the second trimester, patients at term with and without labor, and in patients in preterm labor with and without an intrauterine infection. We compared the amniotic fluid interleukin-10 concentrations among these five groups using the Mann-Whitney U test. RESULTS: Amniotic fluid interleukin-10 was detected in 70% to 91% of patients in each of the five study groups. Higher concentrations were found at term compared with the second trimester (p < 0.001) and concentrations were significantly greater in patients with preterm labor and intrauterine infection compared with those patients in preterm labor without infection (p < 0.001), patients at term in labor (p < 0.001), or patients at term not in labor (p < 0.001). When the patients in preterm labor with infection were analyzed by gestational age, those patients at < 30 weeks had significantly higher amniotic fluid concentrations of interleukin-10 (p = 0.014). CONCLUSIONS: Interleukin-10 was present in the amniotic fluid of the majority of pregnancies, with higher concentrations found at term compared with the second trimester. Intrauterine infection was associated with significantly increased concentrations, with even higher concentrations found in the very premature pregnancies. Interleukin-10 has a prominent yet undefined role in pregnancy and preterm labor complicated by intrauterine infection. PMID- 7485326 TI - Multiple nuchal cord entanglements and intrapartum complications. AB - OBJECTIVE: Our purpose was to evaluate the outcomes of pregnancies complicated by a multiple (double, triple, or quadruple) nuchal cord entanglement. STUDY DESIGN: Computerized data from our University Hospital perinatal database were reviewed between 1990 and 1994. Only singleton, vertex, and term pregnancies undergoing labor were analyzed. Patients with active perinatal complications were eliminated to reduce bias. Pregnancies with infants with either a single or no nuchal cord entanglement served as comparison groups. A comparison of frequencies in the three groups was by chi 2 testing and a comparison of means by a two-tailed Student t test and analysis of variance. RESULTS: Of the 8565 deliveries, the frequency of two or more cord entanglements at delivery was 3.8%. Compared with a single or no cord entanglement, pregnancies with a multiple entanglement were more likely to exhibit an abnormal fetal heart rate pattern during advanced labor (p < 0.001) and to require low or midforceps application (p < 0.001). The study infants were also more likely to have meconium (p = 0.013), a low 1-minute Apgar score (p < 0.001), and an umbilical artery pH < or = 7.10 (odds ratio 2.2, p = 0.013) than the controls. Rates of abruptio placentae, cesarean delivery, and 5 minute Apgar scores < 7 were no more common in the multiple entanglement than the control groups. CONCLUSION: A multiple nuchal cord entanglement was associated with a greater risk of meconium, an abnormal fetal heart rate pattern during advanced labor, the need for operative vaginal delivery, and mild umbilical artery acidosis at birth; however, there was no added risk of an adverse neonatal outcome. PMID- 7485327 TI - The preterm prediction study: significance of vaginal infections. National Institute of Child Health and Human Development Maternal-Fetal Medicine Units Network. AB - OBJECTIVE: Our purpose was to evaluate the association of bacterial vaginosis, trichomonas vaginitis, and monilial vaginitis with spontaneous preterm birth at < 35 weeks 0 days. STUDY DESIGN: A total of 2929 women at 10 centers were studied at 24 and 28 weeks' gestation by Gram stain of vaginal smear, wet mount, and 10% potassium hydroxide preparations to detect vaginal infections. RESULTS: The rates of detected infection at 24 and 28 weeks, respectively, were bacterial vaginosis 23.4% and 19.4%, trichomonas 3.3% and 2.7%, and monilia 21.1% and 19.5%. The occurrence of bacterial vaginosis at 28 weeks was associated with an increased risk of spontaneous preterm birth, odds ratio 1.84 (95% confidence interval 1.15 to 2.95, p < 0.01). Detection of Trichomonas vaginalis (by wet mount) or monilia (by potassium hydroxide preparation) had no significant associations with preterm birth. CONCLUSION: The presence of bacterial vaginosis at 28 weeks' gestation is associated with an increased risk of spontaneous preterm birth. PMID- 7485328 TI - The management of a persistent adnexal mass in pregnancy. AB - OBJECTIVE: Our purpose was to evaluate the pathologic features and outcome of pregnancy complicated by a persistent adnexal mass that was managed conservatively or with surgical intervention. STUDY DESIGN: A review was performed of patients who were seen with an adnexal mass in pregnancy from January 1988 to June 1994. We included patients with simple or complex masses > or = 6 cm that were persistent on ultrasonographic evaluation. We excluded cysts that spontaneously resolved by 16 weeks' gestation and those diagnosed after delivery. RESULTS: Thirty-one patients of 43,372 deliveries were identified with persistent adnexal masses that met the above criteria. Nineteen (59%) of these patients had operative intervention, whereas 12 (41%) were managed conservatively. Of the patients who had surgery, nine had functional cysts, six had mature cystic teratomas, and four had other benign cysts. Complications within 12 hours of surgery included one spontaneous abortion and one patient with rupture of membranes. Twelve patients were managed nonsurgically. Seven patients had conservative management, whereas five patients had percutaneous drainage of simple cysts (negative results on cytologic study) that were symptomatic. CONCLUSIONS: Although the incidence of ovarian cancer in pregnancy is low, the incidental finding of an adnexal mass in pregnancy is becoming more common. Because complications of abdominal surgery are increased in pregnancy, surgical management of this prenatal complication needs to be reconsidered. Our data support a randomized clinical study to determine optimal management of an adnexal mass in pregnancy. PMID- 7485329 TI - Pharmacokinetics and pharmacodynamics of subcutaneous heparin during the early third trimester of pregnancy. AB - OBJECTIVE: Our purpose was to study the pharmacokinetics and pharmacodynamics of heparin administered subcutaneously during the early third trimester of pregnancy. STUDY DESIGN: We gave subcutaneous heparin (143 units/kg) to six pregnant women (mean gestational age 27.1 +/- 1.2 weeks) and to six women who were not pregnant, who served as controls. We serially measured plasma heparin concentrations and activated partial thromboplastin times over 10 hours and compared the time courses of these two measures in the two groups. We calculated the mean peak plasma heparin concentration, time to peak plasma heparin concentration, peak activated partial thromboplastin time, and time to peak activated partial thromboplastin time and compared these variables in the two groups. RESULTS: Plasma heparin concentrations changed significantly over time in both pregnant and nonpregnant subjects. However, the peak plasma heparin concentration in pregnant subjects was significantly lower than in controls (0.11 +/- 0.017 units/ml vs 0.23 +/- 0.036 units/ml) and the time to peak plasma heparin concentration was significantly shorter in the pregnant patients compared with the nonpregnant controls (113 +/- 20 minutes vs 222 +/- 20 minutes). The peak activated partial thromboplastin time (30.3 +/- 1.7 seconds) was significantly lower and time to peak activated partial thromboplastin time (137 +/- 31 minutes) significantly shorter in the pregnant women compared with nonpregnant controls (50 +/- 4.0 seconds; 230 +/- 26 minutes). CONCLUSIONS: Subcutaneous administration of heparin to pregnant patients results in lower plasma heparin concentrations and clinically insignificant prolongation of the activated partial thromboplastin time compared with equivalent doses administered to nonpregnant women. These findings have important implications for the dosing and monitoring of subcutaneous heparin in pregnant women. PMID- 7485330 TI - The effect of magnesium sulfate on bleeding time in pregnancy. AB - OBJECTIVE: The bleeding time is one of the most commonly used diagnostic tests to evaluate platelet-related hemorrhagic disorders. Magnesium has been shown in vitro to be a platelet antiaggregant. This study was conducted to evaluate the hypothesis that magnesium sulfate has no effect on the template bleeding time. STUDY DESIGN: The study group consisted of 24 women who required magnesium sulfate in pregnancy. A blood cell count, platelet count, magnesium level, bleeding time, and mean arterial pressure were obtained before and 2 hours after magnesium sulfate infusion. Magnesium sulfate was infused beginning with a 6 gm intravenous bolus followed by 2 gm/hr. A template bleeding time was performed with a Simplate-II (Organon Teknika, Durham, N.C.) device. Two of the authors performed all the bleeding times. Data were analyzed with a paired t test and Wilcoxon rank test. RESULTS: Fifteen (63%) patients received magnesium sulfate for tocolysis or before external cephalic version and nine (37%) for preeclampsia prophylaxis. No differences were found between the normotensive and hypertensive groups regarding maternal age, gestational age, initial bleeding time, or platelet count. Analysis of the entire study group revealed a prolongation of the bleeding time after magnesium sulfate (5.7 +/- 1.8 vs 6.6 +/- 1.9 minutes, p < 0.05); a lowering of the mean arterial pressure (p < 0.05), and a rise in the magnesium level (p < 0.05). Four patients (16.7%) had a postmagnesium bleeding time > 9 minutes. CONCLUSION: Magnesium sulfate appears to prolong the bleeding time in pregnancy. The clinical significance remains to be determined. PMID- 7485332 TI - Isolated hyperechoic fetal bowel: significance and implications for management. AB - OBJECTIVE: The objective of this study was to determine the significance of isolated hyperechoic fetal bowel. STUDY DESIGN: Forty-five cases with prospective, ultrasonographic diagnosis of isolated hyperechoic fetal bowel were reviewed. Fetal variables, including aneuploidy, deoxyribonucleic acid studies for cystic fibrosis, congenital infection, growth retardation, and intrauterine death were reported. RESULTS: Thirty-four of the 45 cases (76%) resulted in live born infants without detected abnormalities. However, hyperechoic bowel was associated with cystic fibrosis in two cases (4%), congenital infection in two cases (4%), and fetal alcohol syndrome in one case. Termination of pregnancy was elected in three cases and intrauterine fetal death occurred in three cases (7%). Growth retardation was observed in five of 39 (13%) live-born infants. CONCLUSION: Isolated hyperechoic fetal bowel is associated with significant pathologic disorders. Women whose fetuses are diagnosed as having isolated hyperechoic bowel should be offered additional prenatal diagnostic options, including maternal serologic studies for congenital infection, fetal karyotype, and deoxyribonucleic acid testing for cystic fibrosis. In addition, continuing ultrasonographic evaluation of fetal growth and antenatal biophysical assessment should be considered. PMID- 7485331 TI - Favorable hemodynamic effects of magnesium sulfate in preeclampsia. AB - OBJECTIVE: Our purpose was to evaluate the hemodynamic effects of magnesium sulfate in preeclamptic and preterm labor patients. STUDY DESIGN: Fifteen preeclamptic patients at 32.4 +/- 3.3 (mean +/- SD) weeks' gestation and 11 preterm labor patients (31.3 +/- 2.8 weeks) were hemodynamically monitored in the lateral recumbent position by thoracic electrical bioimpedance (BoMed, Irvine, Calif.) before and during high-dose magnesium sulfate bolus and infusion. Cardiac index, systemic vascular resistance index, mean arterial pressure, heart rate, and stroke index were all recorded at baseline and during magnesium sulfate infusion. All patients received a standard crystalloid infusion. Data analysis continued until the patient received epidural placement or other antihypertensive therapy or was delivered. RESULTS: At baseline the systemic vascular resistance index was 2465 +/- 718 F.ohm/m2 and the cardiac index was 3.6 +/- 1.0 L/min/m2 for the preeclamptic patients. These were significantly different compared with the preterm labor patients, who had a systemic vascular resistance index of 1377 +/- 563 F.ohm/m2 and 4.6 +/- 1.2 L/min/m2. Magnesium sulfate infusion resulted in a rapid, sustained fall in systemic vascular resistance and a rise in cardiac index in the preeclamptic patient. This effect was evident at least 4 hours after initiation of the bolus and infusion. In the preterm labor patients the hemodynamic effects of magnesium sulfate were minimal and were noted only during the magnesium sulfate bolus. CONCLUSION: Magnesium sulfate infusion appears to have a prolonged hemodynamic effect in the preeclamptic patient. Sustained reduction in systemic vascular resistance and an increase in cardiac index is found in patients with preeclampsia but not in preterm labor. PMID- 7485333 TI - Fetal fibronectin, interleukin-6, and C-reactive protein are useful in establishing prognostic subcategories of idiopathic preterm labor. AB - OBJECTIVE: Our purpose was to evaluate fetal fibronectin, interleukin-6, and C reactive protein from patients with preterm labor to establish prognostic subcategories. STUDY DESIGN: Thirty-seven patients with preterm labor had cervical fetal fibronectin and plasma C-reactive protein sampled. Eighteen of these patients had amniotic fluid interleukin-6 levels measured. Outcome variables were (1) delivery before 34 weeks and (2) delivery within 48 hours. RESULTS: Detectable cervical fetal fibronectin identified 89% of patients who were delivered before 34 weeks' gestation. Interleukin-6 > 1500 pg/ml identified 88% of patients who were delivered within 48 hours. C-reactive protein > 1.5 mg/dl correlated with elevated interleukin-6 levels (p < 0.001). CONCLUSIONS: Three subcategories of idiopathic preterm labor were evident: (1) fetal fibronectin nondetectable (37% likely to be delivered before 34 weeks), (2) fetal fibronectin detectable but interleukin-6 < 1500 pg/ml (79% likely to be delivered before 34 weeks but 85% with > 48 hours' latency), and (3) fetal fibronectin present and interleukin-6 > 1500 pg/ml (91% likely to be delivered with < 48 hours' latency). PMID- 7485334 TI - Fetal pyelectasis: is it always "physiologic"? AB - OBJECTIVE: Our purpose was to determine the degree of fetal pyelectasis predictive of neonatal renal pathologic processes. STUDY DESIGN: Eighty-four cases of pyelectasis were identified during the study period (1989 through 1993). Fetuses with anteroposterior diameter of the renal pelvis > or = 4 mm before 33 weeks or > or = 7 mm after 33 weeks without caliectasis were included. Fetuses with an anteroposterior diameter of > 10 mm were excluded. Postnatal evaluation included renal sonogram, voiding cystourethrogram, and renal flow and function studies. RESULTS: Sixteen cases were excluded because of incomplete postnatal workup. Renal pathologic processes were found in 30 of 68 (44%) at birth; the most common were ureteropelvic junction obstruction (37%) and vesicoureteral reflux (33%). Only four cases required surgical intervention (13%), and the remaining 87% were managed conservatively. A normal urinary tract was found in 25% of the infants and the remaining 21 of 68 (31%) had regression of pyelectasis before birth. Fetuses with a dilated anteroposterior diameter > or = 8 mm after 28 weeks' gestation were found to have renal pathologic features in two of three of the cases, with sensitivity, specificity, and positive and negative predictive values of 87%, 41%, 66.7% and 70%, respectively. CONCLUSION: Fetuses with an anteroposterior diameter of the renal pelvis > or = 8 mm after 28 weeks' gestation require appropriate urologic evaluation after birth. PMID- 7485335 TI - Precision of office-based blood glucose meters in screening for gestational diabetes. AB - OBJECTIVE: Is the precision of currently available glucose meters adequate for gestational diabetes screening? STUDY DESIGN: We performed a prospective cohort study of 62 gravid women and compared the precision of three glucose meters to laboratory standard technology. RESULTS: The HemoCue coefficient of variation was sufficiently low in venous whole blood and plasma and capillary whole blood and plasma (3.0%, 1.8%, 2.8%, and 2.3%, respectively) to allow use in gestational diabetes screening. The OneTouch II coefficient of variation was sufficiently low in capillary whole blood (2.7%) to allow use in gestational diabetes screening but was too high in venous whole blood and plasma and in capillary plasma (3.6%, 7.8%, and 5.7%). The ExacTech coefficient of variation was too high in venous or capillary whole blood (5.4% and 8.1%) for use in gestational diabetes screening and does not analyze plasma samples. CONCLUSION: The HemoCue meter in venous and capillary whole blood and plasma and the OneTouch II meter in capillary whole blood have coefficients of variation low enough for gestational diabetes screening. PMID- 7485336 TI - Incidence of adverse cardiopulmonary effects with low-dose continuous terbutaline infusion. AB - OBJECTIVE: Our purpose was to determine the incidence of adverse cardiovascular effects of terbutaline sulfate when administered as a continuous subcutaneous infusion in women with arrested preterm labor. STUDY DESIGN: Over a 6-year period records from 8709 women prescribed this therapy for preterm labor that had previously been arrested with other intravenous tocolytics were reviewed. These women were assessed daily for cardiovascular complaints and tolerance of the medication, while either in the hospital or at the home (by telephone). The main outcomes studied were the occurrence of pulmonary edema, sustained cardiac arrhythmias, chest pain, or myocardial ischemia. Any maternal death regardless of cause was also reviewed. RESULTS: Of the 8709 subjects, 47 (0.54%) had one or more cardiopulmonary problems. Pulmonary edema developed in 28 patients (0.32%) while receiving continuous subcutaneous infusion of terbutaline, 5 at home and 23 in the hospital. Of the total, 17 women were being treated concurrently with large amounts of intravenous fluids and one to three other tocolytic agents. In the 11 remaining subjects, 4 were diagnosed with pregnancy-induced hypertension and/or multiple gestation. Nineteen patients experienced other adverse cardiovascular effects, including electrocardiogram changes, irregular heart rate, chest pain, or shortness of breath. CONCLUSIONS: Continuous terbutaline infusion for women with stabilized preterm labor is associated with much fewer adverse effects than previous literature regarding intravenous beta-adrenergic agonist therapy would suggest. PMID- 7485337 TI - Comparison of transcervical and transabdominal chorionic villus sampling loss rates in nine thousand cases from a single center. AB - OBJECTIVES: Our purposes were (1) to compare the safety of transabdominal and transcervical chorionic villus sampling with the use of a consistent technique at one center and (2) to determine whether the training of fellows can be accomplished without an increase in the loss rate. STUDY DESIGN: We performed a retrospective comparison of transabdominal and transcervical chorionic villus sampling loss rates from procedures performed by three principal operators between 1984 and 1992. The type of procedure was chosen by the operator at the time of the procedure on the basis of placental location. RESULTS: Procedures 1 through 2573 were performed solely by transcervical chorionic villus sampling and had an overall fetal loss rate of 5.12%. With the addition of transabdominal chorionic villus sampling the overall fetal loss rate dropped to 3.07% (p < 0.0001). Three and one half years after the start of transabdominal chorionic villus sampling (about 1300 transabdominal chorionic villus sampling procedures), the transabdominal chorionic villus sampling loss rate was significantly better than the transcervical loss rate (p = 0.035), and the difference widened steadily after that. During the same time period seven fellows performed 716 procedures for a fetal loss rate among fellows of 2.72%. CONCLUSIONS: (1) Under optimal circumstances (one center, large numbers, few operators, consistent technique, operator choice of best approach), transabdominal chorionic villus sampling may be inherently safer than transcervical chorionic villus sampling. (2) The addition of transabdominal chorionic villus sampling decreases overall chorionic villus sampling loss rates. (3) Although the number of procedures performed by fellows is small, it appears that with close supervision by experienced operators successful training of fellows can be accomplished without adverse effects on loss rates. PMID- 7485338 TI - Detection of viral deoxyribonucleic acid in the amniotic fluid of low-risk pregnancies by polymerase chain reaction. AB - OBJECTIVE: The purpose of this study was to determine whether viral deoxyribonucleic acid is detectable in the amniotic fluid of pregnancies at low risk for fetal viral infection. STUDY DESIGN: Amniotic fluid samples were prospectively collected from 277 patients. Selected primer pairs amplified deoxyribonucleic acid sequences unique to adenovirus, cytomegalovirus, herpes simplex virus, and parvovirus. Amplified deoxyribonucleic acid was detected by gel electrophoresis. Sensitivity of the adenovirus, cytomegalovirus, and herpes virus primers were determined by serial dilution of 10(3) PFU/ml controls. RESULTS: Of the 277 extracted samples, 243 had detectable deoxyribonucleic acid. None of these samples had detectable viral deoxyribonucleic acid by polymerase chain reaction. The sensitivity of the adenovirus primer pairs was 10(-3) PFU/ml, cytomegalovirus 10(-2) PFU/ml, and herpes simplex virus 10(-1) PFU/ml. CONCLUSION: This study did not detect viral deoxyribonucleic acid in a low-risk population, supporting the clinical significance of detecting viral deoxyribonucleic acid in pregnancies at risk for infection. PMID- 7485339 TI - Comparison of multiple-marker screening with amniocentesis for the detection of fetal aneuploidy in women > or = 35 years old. AB - OBJECTIVE: Our purpose was to compare the multiple-marker screening test with elective amniocentesis for the detection of fetal Down syndrome and other aneuploidies in women aged > or = 35. STUDY DESIGN: Our database included the multiple-marker screening test (maternal serum alpha-fetoprotein, human chorionic gonadotropin, unconjugated estriol, and maternal age) and genetic amniocentesis results from 1942 women aged > or = 35. A Down syndrome risk > or = 1:190 was considered screen positive. An algorithm to detect trisomy 18 was also used. RESULTS: The multiple-marker screening test Down syndrome screen-positive rate was 26.1% (507/1942). The Down syndrome detection rate was 75% (33/44); the trisomy 18 detection rate was 75% (3/4). However, the multiple-marker screening test detection rate for all aneuploidies was only 61%. Missed aneuploidies included trisomy 21, sex chromosome abnormalities, trisomy 13, trisomy 22, and trisomy 18. CONCLUSIONS: The multiple-marker screening test fails to detect approximately 39% of all fetal aneuploidies in women aged > or = 35. These data should be provided to women considering prenatal diagnosis so that they can make an informed decision regarding the multiple-marker screening test versus amniocentesis for advanced maternal age. PMID- 7485341 TI - The influence of maternal weight on human chorionic gonadotropin in the multiple marker screening test for fetal Down syndrome. AB - OBJECTIVE: Our purpose was to determine the effect of maternal weight on human chorionic gonadotropin concentration in the multiple-marker screening test for fetal Down syndrome. STUDY DESIGN: Two genetics databases were used: database I contained the results of 8297 multiple-marker screening tests and database II contained the results of 1936 multiple-marker screening tests and fetal karyotypes. RESULTS: The overall screen-positive rate in database I was 7.1%; it was 7.5% in patients weighing < 180 pounds and 5.1% in patients weighing > or = 180 pounds (p = 0.001). Weight significantly affected the screen-positive rate only in women > or = 30 years old (p = 0.003 for 30 to 34 years, p = 0.00004 for > or = 35 years). A weight correction formula was derived; when applied to database II it eliminated individual weight-related differences but had no effect on the overall screen-positive rate or Down syndrome detection rate. CONCLUSIONS: Human chorionic gonadotropin concentration is affected by maternal weight. A weight correction formula eliminates individual weight-related differences in the screen-positive rate but has no discernible effect on the overall screen-positive or Down syndrome detection rates. PMID- 7485340 TI - The effect of low-dose aspirin on pregnancies complicated by elevated human chorionic gonadotropin levels. AB - OBJECTIVE: Our purpose was to determine whether elevated second-trimester human chorionic gonadotropin levels identify women likely to benefit from low-dose aspirin therapy. STUDY DESIGN: We evaluated second-trimester human chorionic gonadotropin levels obtained from healthy nulliparous women before screening for participation in a double-blind randomized trial of aspirin therapy: 262 women took 60 mg of aspirin daily and 420 did not. RESULTS: Among women who did not take aspirin, those with human chorionic gonadotropin levels > or = 2.0 multiples of the median had a significantly lower mean birth weight (2859 vs 3159 gm, p = 0.04) than did those with normal human chorionic gonadotropin levels. All women who took aspirin had a higher mean birth weight than women who did not, but women with human chorionic gonadotropin levels > or = 2.0 multiples of the median had the greatest increase (416.2 gm higher in those with human chorionic gonadotropin levels > or = 2.0 multiples of the median, p = 0.02; 96 gm higher in those with human chorionic gonadotropin levels > or 2.0 multiples of the median, p = 0.04). Regression analysis suggested that the higher birth weight was partly explained by a higher gestational age at delivery and partly by increased weight independent of gestational age. CONCLUSIONS: Aspirin therapy increased birth weight in all women, especially in women with high human chorionic gonadotropin levels, partly by increasing gestational age at delivery. This observation needs to be confirmed by further studies. PMID- 7485342 TI - Inhibition of neutrophil oxidative burst and phagocytosis by meconium. AB - OBJECTIVE: Meconium in amniotic fluid has been associated with an increased prevalence of chorioamnionitis. In an effort to delineate the mechanism of this association, we determined the effect of meconium on the neutrophil's capacity for phagocytosis and microbial killing by oxidative burst in vitro. STUDY DESIGN: Sterile meconium samples were obtained from four fetuses at the time of breech delivery and were then pooled and lyophilized. Neutrophils were purified from whole blood of each of 13 pregnant nonlaboring patients. Phagocytosis and the oxidative burst of neutrophils in the presence and absence of meconium were assessed by single-cell analysis with flow cytometry. Phagocytosis was measured as the mean fluorescence intensity produced after 30 minutes of incubation with fluorescein-labeled Escherichia coli. Oxidative burst was measured as the mean fluorescence intensity resulting from the oxidation of internalized reduced dichlorodihydrofluorescein after 15 minutes of stimulation with phorbol myristate acetate. Oxidative burst was expressed as the neutrophil oxidative index and the net fluorescence intensity. Neutrophil oxidative index was equivalent to the quotient of the mean fluorescence intensity for phorbol myristate acetate stimulated and unstimulated cells. Net fluorescence intensity was equivalent to the absolute difference between stimulated and unstimulated cells. RESULTS: Exposure of neutrophils to light and very light meconium each resulted in significantly lower mean neutrophil oxidative index compared with unexposed controls (3.2 +/- 4.9 and 4.2 +/- 5.9 vs 16.2 +/- 7.5, p = 0.00002 and p = 0.0007, respectively) and significantly lower mean net fluorescence intensity than that of control cells (112 +/- 220 and 188 +/- 294 vs 613 +/- 328, p = 0.0001 and p = 0.005, respectively). Phagocytosis was significantly impaired in the presence of moderate meconium compared with control cells (2239 +/- 393 vs 4645 +/- 2071, p = 0.0001). Light meconium did not significantly affect phagocytosis. CONCLUSION: Meconium has significant effects on neutrophil function in vitro. Both light and very light meconium inhibit the oxidative burst. Moderate meconium inhibits phagocytosis. PMID- 7485343 TI - Triple marker (alpha-fetoprotein, unconjugated estriol, human chorionic gonadotropin) versus alpha-fetoprotein plus free-beta subunit in second-trimester maternal serum screening for fetal Down syndrome: a prospective comparison study. AB - OBJECTIVE: Our purpose was to compare the efficacy of triple-marker screening (alpha-fetoprotein, unconjugated estriol, human chorionic gonadotropin) with alpha-fetoprotein plus free beta-human chorionic gonadotropin. STUDY DESIGN: Free beta-human chorionic gonadotropin was concurrently assayed in 2349 maternal serum samples. Trivariate and bivariate algorithms were used to calculate the risk for fetal Down syndrome by the two protocols. Free beta-human chorionic gonadotropin from 12 cases of fetal Down syndrome previously screened with the triple marker was retrospectively assayed. RESULTS: Mean maternal age of our study was 29.8 years (range 14 to 51 years). The initial screen-positive rate with the triple marker was 8.0% compared with 12.8% for alpha-fetoprotein plus free beta-human chorionic gonadotropin. All three cases of fetal Down syndrome ascertained in our prospective study were detected by the triple marker; in contrast, one of three was detected by alpha-fetoprotein plus free beta-human chorionic gonadotropin. By adding 12 additional cases of fetal Down syndrome, 12 of 15 (80%) were screen positive with triple marker and nine of 15 (60%) were screen positive with alpha fetoprotein plus free beta-human chorionic gonadotropin. CONCLUSION: The detection rate of fetal Down syndrome was greater by use of a triple marker screen than when using alpha-fetoprotein plus free beta-human chorionic gonadotropin. Our data do not support the claims of other studies that suggest that alpha-fetoprotein plus free beta-human chorionic gonadotropin is superior to triple markers. PMID- 7485344 TI - Risk factors for the development of preterm premature rupture of the membranes after arrest of preterm labor. AB - OBJECTIVE: Our purpose was to determine risk factors predictive of preterm premature rupture of the membranes in women treated for preterm labor with intact membranes. STUDY DESIGN: Women with intact membranes participating in a National Institute of Child Health and Human Development multicenter randomized trial of adjunctive antibiotic therapy for preterm labor (24 to 34 weeks) were studied (n = 275). After randomization, 22 women continued to have contractions and were delivered of their infants. The remaining 253 women whose contractions had ceased composed our study population. Preterm premature rupture of the membranes was diagnosed if ruptured membranes occurred > or = 1 hour before the onset of recurrent preterm labor. As part of the study protocol, most women underwent amniocentesis on admission. RESULTS: Preterm premature rupture of the membranes developed in 44% women (17.4%). Women who had preterm premature rupture of the membranes were more likely to be black (p = 0.004), to be multiparous (p = 0.014), to have a history of abortion(s) (p = 0.001), to have had a preterm birth(s) (p = 0.036), to have early onset preterm labor (p = 0.04), to have more advanced cervical dilatation (p = 0.0001), to have one or more amniotic fluid markers suggestive of infection (p = 0.01, odds ratio 4.2), and to have positive amniotic fluid cultures (p = 0.0007, odds ratio 27). Assignment to antibiotic therapy did not prevent preterm premature rupture of the membranes in the 253 women randomized or in the 16 women with a positive amniotic fluid marker(s) of infection. CONCLUSION: Black race, multiparity, a history of abortion or preterm birth, advanced dilatation, and a positive amniotic fluid marker(s) are associated with preterm premature rupture of the membranes in women with preterm labor. Antibiotic treatment did not prevent preterm premature rupture of the membranes. PMID- 7485345 TI - Premature parturition is characterized by in utero activation of the fetal immune system. AB - OBJECTIVE: At birth the fetus emerges from a sterile environment into a nonsterile one. This process is associated with activation of the fetal immune system which protects the fetus against infection in the newborn period. We conducted this study to determine whether activation of the monocyte-neutrophil system occurs in fetuses before premature birth. STUDY DESIGN: Forty patients in premature labor with intact membranes underwent cordocentesis for research purposes. Fetal blood was analyzed with the use of flow cytometry to measure the cell surface markers CD11c, CD13, CD15, and CD67, which are associated with monocyte and neutrophil activation, and CD14 and CD63, which were used as controls. RESULTS: Twenty-eight percent (11/40) of the infants were delivered prematurely within 72 hours of entering the study while the remainder were delivered at term. Our data clearly indicate that premature infants delivered within 72 hours had a higher percentage of CD11c, CD13, CD15, and CD67 than those delivered at term. In contrast, there were no significant differences in the percentages of CD14 and CD63. CONCLUSION: Activation of the monocyte-neutrophil system exists in fetuses destined for premature delivery. These findings indicate that premature parturition is associated with in utero immune system activation. PMID- 7485349 TI - Prenatal cytogenetic abnormalities: correlations of structural rearrangements and ultrasonographically detected fetal anomalies. AB - OBJECTIVE: Our purpose was to determine the distribution of karyotypic abnormalities detected at prenatal diagnosis, fetal anomalies, and ability for fluorescent in situ hybridization detection. STUDY DESIGN: Our cytogenetic database from January 1988 to April 1994 was categorized according to type and potential detection by current standard fluorescent in situ hybridization probes. Fetal anomalies and cytogenetic aberrations were compared. RESULTS: A total of 664 cases of abnormal fetal karyotypes were identified from 12,454 prenatal cytogenetic cases (7529 amniocenteses and 4925 chorionic villus sampling) and were classified as autosomal aneuploidy (331), sex aneuploidy (103), polyploidy (38), marker aneuploidy (19) and structural rearrangements (173). Standard fluorescent in situ hybridization probes would have missed 31% of the abnormal cases: 90 aneuploidy, 14 de novo marker aneuploidy, and 65 de novo structural aberrant cases. The 134 cases of structural chromosomal rearrangements with complete ultrasonographic records were further classified as polymorphism (42), familial (43), or de novo (49). Frequency of fetal anomaly detection by ultrasonography in de novo cases (22/49) was higher than other rearrangements (chi 2 7.4, p = 0.006). CONCLUSION: The contribution of unusual aneuploidies (16%) and structural chromosomal rearrangements (26%) in prenatal diagnostic practice is significant. Fetal anomalies were detected by ultrasonography in 45% of the de novo rearrangement cases. Fluorescent in situ hybridization would miss 31% of the abnormal cases. PMID- 7485347 TI - The accuracy and patient preference for self-collected group B Streptococcus cultures. AB - OBJECTIVE: Our purpose was to determine the accuracy of and patient attitudes regarding self-collected group B Streptococcus cultures. STUDY DESIGN: Women seen for prenatal care at 24 to 42 weeks' gestation were asked to collect distal vaginal and anal samples for group B Streptococcus. Subsequently, distal vaginal and anal samples were obtained by the nurse. The patients were then asked their preference toward self-sampling. RESULTS: A total of 251 women participated in the study. The incidence of positive group B Streptococcus cultures was 12.7%, 9.6%, 10.0%, and 7.6% for the patient-collected vaginal and anal and nurse collected vaginal and anal specimens, respectively. The incidence of group B Streptococcus carriage was 17.5% and 13.5% for any positive patient- or provider collected specimens, respectively, and 19.1% for any positive culture. Single patient-collected vaginal and anal and nurse-collected vaginal and anal samples were insensitive for group B Streptococcus carriage (67%, 50%, 52%, 40%, respectively). The combination of patient-collected samples was more sensitive than nurse-collected samples (sensitivity 91.7% vs 70.8%, p < 0.05). Repeat sampling of the vagina or anal canal did not offer significant additional benefit to a single culture. Overall, patient-collected samples were 98.4% accurate in predicting group B Streptococcus carriage versus 94.4% for nurses. A total of 58% of women preferred obtaining their own specimens, whereas 9.6% found the technique difficult. Ninety percent desired the option of self-sampling in the future. CONCLUSIONS: Single vaginal or anal cultures were insensitive in detecting group B Streptococcus carriage. Combined patient-collected cultures were more sensitive than provider-collected specimens. On the basis of accuracy and patient preference, women should be given the opportunity of combined vaginal anal self-sampling for group B Streptococcus when indicated. PMID- 7485348 TI - The impact of initiating a human immunodeficiency virus screening program in an urban obstetric population. AB - OBJECTIVE: Our purpose was to describe the incidence of human immunodeficiency virus infection and to assess the cost/benefit ratio of universal antenatal human immunodeficiency virus screening. STUDY DESIGN: Medical records of women in this urban obstetrics population, from the years 1988 to 1993, were examined. The incidence of known human immunodeficiency virus seropositivity at delivery was determined. The costs of performing human immunodeficiency virus screening, evaluating the disease status, and administering therapy were calculated. These costs were compared with an averaged cost for care and follow-up of infants infected through vertical transmission. RESULTS: The incidence of known human immunodeficiency virus seropositivity at delivery approximately doubled since the initiation of a human immunodeficiency virus screening program (0.26% to 0.48%). Obstetric screening added an approximate $100,000 to medical costs. The calculated cost of pediatric follow-up of human immunodeficiency virus seropositive infants for the first 18 months was estimated at $344,355. In our population, with universal screening and zidovudine therapy, the medical costs could be reduced by $175,500 per year. CONCLUSION: A program of voluntary human immunodeficiency virus screening increases the incidence of known human immunodeficiency virus infection. Offering screening and follow-up to all pregnant patients in an urban setting is both cost-effective and medically beneficial. PMID- 7485346 TI - Early versus late amniotomy for labor induction: a randomized trial. AB - OBJECTIVE: Our purpose was to determine the impact of early and late amniotomy on labor induction with continuous oxytocin infusion at term. STUDY DESIGN: A total of 209 women admitted for labor induction were randomized to early or late amniotomy. The early amniotomy group (n = 106) had membranes ruptured as soon as it was deemed safe and feasible. The late amniotomy group (n = 103) had membrane rupture performed at > or = 5 cm dilatation. The first 103 women received a continuous oxytocin infusion with incremental adjustments at 60-minute intervals as required. The next 106 women had adjustments every 30 minutes as required. Statistical analysis was confined to concurrent groups. RESULTS: Early amniotomy was associated with shorter labor (13.3 vs 17.8 hours, p = 0.001), chorioamnionitis (22.6% vs 6.8%, p = 0.002), and significant fetal umbilical cord compression (12.3% vs 2.9%, p = 0.017). The benefit regarding shortening of labor was limited to women having oxytocin increments every 30 minutes as required (13.3 vs 17.8 hours, p = 0.001). Alternatively, the increase in chorioamnionitis was confined to the 60-minute group (39% vs 11%, p < 0.001), which also demonstrated a trend toward increased moderate and severe variable decelerations (19.6% vs 6.4%, p = 0.08). CONCLUSIONS: When a protocol of 60-minute increments in oxytocin infusion rate is desired, amniotomy should be performed late in labor to reduce chorioamnionitis and significant umbilical cord compression. Alternatively, if early amniotomy is necessary, oxytocin should be adjusted every 30 minutes as tolerated. PMID- 7485350 TI - Salivary estriol as risk assessment for preterm labor: a prospective trial. AB - OBJECTIVE: Increased understanding of human parturition allows for novel approaches to (1) identification of women at increased risk for preterm birth and (2) development and controlled testing of etiology-based strategies to prevent preterm birth. STUDY DESIGN: Five hundred forty-two women were enrolled at five study sites in a prospective evaluation of salivary estriol in samples obtained weekly beginning at 22 weeks' gestation (Salest, Biex, Inc., Boulder, Colo.). Estriol concentrations were determined with a well-characterized enzyme-linked immunoassay. Women adjudged at either high risk or low risk for prematurity were evaluated through to delivery. RESULTS: A total of 267 women submitted serial samples that were analyzed; 241 women with singleton pregnancies submitted sufficient samples. Twenty-three women with singleton fetuses went into idiopathic preterm labor (without prior rupture of membranes) and were delivered preterm (mean 35 weeks' gestation); 182 were delivered at term (> or = 37 weeks' gestation). Mean (geometric) estriol concentrations were higher from 24 to 34 weeks in women with singleton pregnancies delivering preterm (p < 0.05). A surge in estriol concentrations occurred approximately 3 weeks before the onset of labor in both women delivering at term and those delivering preterm. This increase occurred approximately 4 weeks earlier in women delivered preterm versus term. Receiver-operator curve analyses showed that exceeding a 2.3 ng/ml saliva estriol level was associated with occurrence of preterm labor (71% sensitivity, 77% specificity, 23% false-positive rate). CONCLUSION: Detection of an early estriol surge or increased level (> or = 2.3 ng/ml) may be clinically helpful in identifying women at elevated risk for preterm labor and birth, allowing for evaluation of biologically based interventions in controlled trials. PMID- 7485351 TI - Cerebellar growth in normal and growth-restricted fetuses of multiple gestations. AB - OBJECTIVE: Our purpose was to evaluate growth of the cerebellum in growth restricted fetuses of twin and triplet gestations versus growth in normal in utero sibling(s) and in singleton gestations. STUDY DESIGN: An ultrasonographic study was conducted in a population of pregnant women with twin and triplet gestations. The control group was either the normal in utero sibling(s) when one fetus was growth restricted or normal twin and singleton pregnancies. Standard biometric measurements were obtained on all fetuses throughout pregnancy, including the transverse cerebellar diameter. However, only the last measurement was used for the analysis. Statistical analyses were conducted comparing growth of the transverse cerebellar diameter among the growth-restricted fetuses versus growth in the normal in utero fetal sibling(s) or other normal twin and singleton gestations. RESULTS: Pregnancies were categorized on the basis of the growth status of women with twin and triplet gestations: Group 1 (151) contained women with two fetuses appropriately grown for gestational age; group 2 (52) had one appropriately grown fetus and one with intrauterine growth restriction; group 3 (19) had two fetuses with intrauterine growth restriction. In addition, there were 30 triplet gestations (group 4), five of which had growth-restricted fetuses, and group 5 contained 1405 singleton pregnancies. In all five groups there was a statistically significant relationship between transverse cerebellar diameter and gestational age (p < 0.0001). There was also no significant difference between growth of the transverse cerebellar diameter in the appropriately grown and growth-restricted siblings and among normal singleton and twin pregnancy groups. In most cases of growth-restricted fetuses, except for the transverse cerebellar diameter measurements, all other biometric parameters were < 10th percentile. CONCLUSION: These data confirm the relative preservation of normal cerebellar growth in growth-restricted fetuses and a similar rate of growth in singleton and multifetal gestations. The transverse cerebellar diameter therefore represents an independent biometric parameter that can be used in both singleton and multifetal pregnancies to assess normal and deviant fetal growth. PMID- 7485353 TI - Whitridge Williams' obstetrics. PMID- 7485352 TI - De conceptu et generatione hominis. PMID- 7485354 TI - Ultrasonographic evaluation of patients with abnormal bleeding. PMID- 7485356 TI - Computerized analysis of antepartum fetal heart rate. PMID- 7485355 TI - Value of transvaginal ultrasonography and hysteroscopy in the evaluation of postmenopausal women with metrorrhagia. PMID- 7485357 TI - Prenatal diagnosis by use of fetal cells isolated from maternal blood. PMID- 7485358 TI - Noncontact transscleral Nd:YAG cyclophotocoagulation for glaucoma after penetrating keratoplasty. AB - PURPOSE: To evaluate the effect of noncontact transscleral Nd:YAG cyclophotocoagulation for refractory glaucoma after penetrating keratoplasty on intraocular pressure, visual acuity, and graft status. METHODS: We reviewed the records of all patients treated at the Duke University Eye Center between 1988 and 1993 who had noncontact transscleral Nd:YAG cyclophotocoagulation for uncontrolled glaucoma associated with penetrating keratoplasty. RESULTS: One eye each of 39 patients was included in the study. At final follow-up, ranging from three to 63 months (mean, 27 +/- 18 months), the average decrease in intraocular pressure was 15 mm Hg (P = .001). Thirty-one patients (77%) had a final intraocular pressure between 7 and 21 mm Hg. The number of medications was reduced by an average of 1.4 (P = .0001). Five patients (13%) showed an improvement in best-corrected visual acuity at final follow-up; 12 (31%) maintained stable acuity, and 22 (56%) demonstrated a- deterioration in acuity. Of 25 patients with clear grafts before cyclophotocoagulation, 11 (44%) had graft decompensation. CONCLUSIONS: Noncontact transscleral Nd:YAG cyclophotocoagulation is effective in lowering intraocular pressure in eyes with refractory glaucoma and associated penetrating keratoplasty, although loss of visual acuity and graft decompensation remain important concerns. Further study is needed of other cyclophotocoagulation protocols and alternative treatments for this patient population. PMID- 7485359 TI - Grading of retinal nerve fiber layer with a photographic reference set. AB - PURPOSE: We developed a new quantitative approach for the evaluation of diffuse atrophy of the retinal nerve fiber layer. METHODS: A simultaneous, visually supported grading system was designed, which consisted of a set of 25 reference photographs, numbered from 25 (broad, clearly striated nerve fiber bundles) to 1 (no nerve fibers visible). We prospectively evaluated this method by matching 60 retinal nerve fiber layer photographs of patients with glaucoma or ocular hypertension and normal subjects to the reference photographs twice by three observers with varying experience levels in evaluating retinal nerve fiber layer RESULTS: Intraobserver and interobserver reliability, evaluated by the intraclass correlation coefficient statistics, was excellent (intraclass correlation coefficient > 0.8). Intraclass correlation coefficients within observers were from 0.89 to 0.98 (lower limits 905% confidence interval, 0.84 to 0.97), and intraclass correlation coefficients between observers were from 0.81 to 0.91 (lower limits 95% confidence interval, 0.50 to 0.87). Clinical validity, in which the scores for upper and lower halves of photographs were compared with the mean deviation of the Humphrey 30-2 visual field program by using the Spearman correlation coefficient, was substantial (r = -.68, P < .01 for upper photo score and lower field mean deviation, r = -.53, P < .01 for lower photo score and upper field mean deviation). CONCLUSIONS: By using a reference set of of photographs of the retinal nerve fiber layer, we defined a method to derive a quantitative measurement of retinal nerve fiber layer with good reliability and to extend evaluation of retinal nerve fiber layer photographs to nonspecialists. PMID- 7485360 TI - Random dot motion perimetry in patients with glaucoma and in normal subjects. AB - PURPOSE: To determine whether patients with primary open-angle glaucoma have an increase in size thresholds, prolongation of reaction times, and greater localization errors to random dot motion stimuli than normal subjects. METHODS: Motion perimetry, a computer graphics method of visual field testing, quantitates a subject's ability to detect a correlated shift in position of dots within a defined circular area against a background of fixed dots. We measured motion thresholds, the smallest detectable circular dot motion target, at the Humphrey 24-2 test loci. By using the subject's light-pen responses to the location of the targets, we computed motion size threshold, reaction times, and localization errors (number of pixels from where the subject touched the monitor to the target center). With motion perimetry and conventional automated perimetry, we tested one eye in each of 25 patients with primary open-angle glaucoma and 25 age matched control subjects. We then generated total deviation pointwise probability plots for the patients with primary open-angle glaucoma. RESULTS: Patients with primary open-angle glaucoma had increased mean motion size threshold (P < .001) and increased localization errors (P < .002), compared with the control subjects. With the probability plot analysis, there was good correlation of the visual field defects between the two perimetry tests. Additionally, motion perimetry identified nerve fiber bundle-like defects in 12 patients that were not detected with conventional automated perimetry. CONCLUSIONS: Patients with primary open angle glaucoma had abnormal motion perception with an increase in spatial localization error. PMID- 7485362 TI - Clinical assessment, optic disk measurements, and visual-evoked potential in optic nerve hypoplasia. AB - PURPOSE: Because there is a broad spectrum of visual dysfunction in eyes with optic nerve hypoplasia, we determined whether any clinical observations, including optic disk measurements, could predict visual outcome in eyes with hypoplastic optic nerves. METHODS: We prospectively studied the eyes of 19 children (eight consecutive and 11 randomly selected), ages 7 to 92 months, in whom a diagnosis of optic nerve hypoplasia was made by an ophthalmologist. Clinical assessment included visual acuity, pupillary responses, and nystagmus. The ratio of the horizontal disk diameter to the disk-macula distance was calculated from fundus photographs. The amplitude and latency of the flash visual evoked potential as well as the threshold response to pattern reversal checks were recorded. RESULTS: Visual acuity was associated with pupil reactivity, nystagmus, the ratio of the horizontal disk diameter to the disk-macula distance, pattern visual-evoked potential threshold, and flash visual-evoked potential amplitude (P < .001 for all parameters). All eyes with a ratio of the horizontal disk diameter to the disk-macula distance greater than .30 had good visual acuity, and all eyes with a ratio of the horizontal disk diameter to the disk macula distance less than .30 had poor visual acuity. No eye with a ratio of the horizontal disk diameter to the disk-macula distance less than or equal to .15 had visual acuity better than light perception. CONCLUSIONS: Visual-evoked potential and photogrammetric measurement of relative disk size can be used to approximate visual acuity in eyes with optic nerve hypoplasia. This finding could lead to a diagnostic criterion or a severity classification for optic nerve hypoplasia. PMID- 7485361 TI - Ocular and systemic manifestations of cerebrotendinous xanthomatosis. AB - PURPOSE: Cerebrotendinous xanthomatosis is a storage disease that usually leads to severe mental and neurologic deterioration before the diagnosis and start of treatment are established. We identified major ocular and systemic characteristics that may enable a diagnosis to be made earlier. METHODS: Ten patients (group 1) of the University Hospital Nijmegen, with a diagnosis of cerebrotendinous xanthomatosis, were re-examined for detailed ocular and major clinical manifestations. Meanwhile, we looked for similar but undiagnosed cases in patients (group 2) who visited the Institute of Ophthalmology during a 12 month period. RESULTS: A diagnosis of cerebrotendinous xanthomatosis had been made in the patients of group 1 at an average age of 40 years (range, 33 to 48 years). Subsequently, six new cases (group 2) were diagnosed in patients 7 to 37 years old (average age, 18 years). Bilateral cataract was the major ocular manifestation in all 16 patients. Small irregular corticonuclear opacities, anterior polar cataracts, and dense posterior subcapsular cataracts were diagnosed at various ages (mean, 18 years; range, 4 to 40 years). Four patients showed clinical signs of optic neuropathy, whereas retinal function was normal in all patients. Other major clinical signs included a history of chronic diarrhea (since childhood), mental deterioration (mean age, 23 years), neurologic deterioration (mean age, 31 years), and tendon xanthomas (mean age, 37 years). CONCLUSIONS: Appropriate biochemical investigations for cerebrotendinous xanthomatosis should be performed in patients with unexplained juvenile or early onset adult cataracts, especially if these cataracts are associated with chronic diarrhea since infancy, mental retardation or deterioration, neurologic dysfunction, or xanthomas. PMID- 7485364 TI - Variability of the relative afferent pupillary defect. AB - PURPOSE: Afferent asymmetry of visual function is detectable in both normal and pathologic conditions. With a computerized test, we assessed the variability in measuring afferent asymmetry of the pupillary light reflex, that is, the relative afferent pupillary defect. METHODS: In ten normal subjects, pupillary responses to an alternating light stimulus were recorded with computerized infrared pupillography. The relative afferent pupillary defect for each test was determined by using a new computer analysis. The 95% confidence interval of each determination of relative afferent pupillary defect was used to represent the short-term fluctuation in its measurement. To optimize the test for clinical use, we studied the influence of stimulus intensity, duration, and number on the variability of the relative afferent pupillary defect. RESULTS: When the relative afferent pupillary defect was based on only a few light alternations (stimulus pairs), there was excessive variability in its measurement (95% confidence interval > 0.5 log units). With approximately 200 stimulus pairs, the 95% confidence interval was reduced to less than 0.1 log unit (relative afferent pupillary defect +/- 0.05 log unit). Also, there was less variability when the dark interval between alternating light stimulation was less than one second. CONCLUSIONS: Computerized infrared pupillography can standardize the alternating light test and minimize the error in quantifying a relative afferent pupillary defect. A reproducible relative afferent pupillary defect measurement is desirable for defining afferent injury and following the course of disease. PMID- 7485363 TI - Neuro-ophthalmic manifestations of meningocerebral inflammation from the limited form of Wegener's granulomatosis. AB - PURPOSE: Wegener's granulomatosis is classically a systemic necrotizing granulomatous vasculitis that involves the upper and lower respiratory tracts and kidneys. Ophthalmologic and neurologic manifestations are common. The limited form of Wegener's granulomatosis may have pathologic characteristics consistent with the disease but lacks the complete clinical triad. We studied the clinical, pathologic, laboratory, and radiologic findings of four patients with Wegener's granulomatosis so that others will recognize the disease, even when it occurs in its limited form. METHODS: From three clinical centers, a chart review disclosed four patients with neuro-ophthalmic findings and the limited form of Wegener's granulomatosis. RESULTS: Three men and one woman, ranging in age from 40 to 73 years, were studied. All four patients had ocular motor abnormalities (one with oculomotor and trochlear nerve palsies, one with oculomotor nerve palsy, one with trochlear and abducens nerve palsies, and one with horizontal gaze deviation) in addition to other cranial nerve and cerebral abnormalities. Neuroimaging showed prominent meningeal, as well as intraparenchymal, abnormalities. In all patients, results of antineutrophil cytoplasmic antibody tests were initially negative but in one patient were positive at a late stage of the disease. In all patients, results of a biopsy demonstrated necrotizing granulomatous inflammation consistent with Wegener's granulomatosis. CONCLUSIONS: Neuro-ophthalmic findings may be the earliest manifestations of the limited form of Wegener's granulomatosis. Extensive meningocerebral inflammation can occur before systemic involvement or laboratory confirmation. Early diagnosis by biopsy of affected tissues may facilitate appropriate treatment and prevent progression of the disease. PMID- 7485365 TI - Ocular abnormalities in neurofibromatosis 2. AB - PURPOSE: To evaluate the ocular abnormalities in patients with clinically diagnosed neurofibromatosis 2 and asymptomatic gene carriers. METHODS: Probands were ascertained through a surgical otolaryngology practice. In a cross-sectional study, we examined 49 patients with neurofibromatosis 2, 30 offspring of patients, and, as a comparison group, 18 parents and siblings of patients with sporadic neurofibromatosis 2. The examination included a complete neuro ophthalmic assessment, physical examination, and, for patients and first-degree relatives at risk, cranial and spinal magnetic resonance imaging with gadolinium enhancement, if not previously performed. RESULTS: The most common ocular abnormalities were posterior subcapsular or capsular, cortical, or mixed lens opacities in 33 (67%) of 49 patients with neurofibromatosis 2 and retinal hamartomas in 11 (22%). We used segregation analysis to determine the mutation carrier status of six at-risk offspring who were 30 years old or younger in two multigeneration families. Three asymptomatic mutation carriers had cataracts, whereas those who were predicted not to carry the mutation did not have cataracts. Asymptomatic mutation carriers may have developmental abnormalities of the eye that are detectable in childhood or adolescence, a finding that may assist in early diagnosis of the disease. CONCLUSIONS: A variety of ocular abnormalities are present in neurofibromatosis 2, including cataracts, retinal hamartomas, and ocular motor deficits. Many of these are developmental or acquired early in life and may assist in presymptomatic diagnosis. For screening at-risk relatives of patients with neurofibromatosis 2, the types of cataract that are most suggestive of neurofibromatosis 2 are plaque-like posterior subcapsular or capsular cataract and cortical cataract with onset under the age of 30 years. PMID- 7485366 TI - Confocal scanning infrared laser ophthalmoscopy for indocyanine green angiography. AB - PURPOSE: We used indocyanine green to study wavelength-optimized confocal scanning infrared laser angiography in patients with retinal and choroidal disease. METHODS: A confocal scanning laser ophthalmoscope with an excitation wavelength of 795 nm was operated both in tight and wide confocal imaging modes. We examined 77 subjects with and without retinal and choroidal disease (including diabetic retinopathy, age-related macular degeneration, and subretinal neovascularization). RESULTS: The scanning laser ophthalmoscope allowed acquisition of images, in the wide confocal imaging mode, of the retinal circulation and late leakage sites without late injections of dye to outline the retinal vasculature. In the tight confocal imaging mode, optical subtraction of the light contribution of the retinal circulation allowed examination of the choroidal circulation, and vice versa. The wide confocal mode appears equivalent to other scanning laser ophthalmoscopes in recording images from retinal and choroidal layers. CONCLUSIONS: There are three differences between the confocal scanning laser ophthalmoscope and conventional instruments. First, the late images allow excellent visualization of the retinal circulation without a landmark injection. Second, confocal imaging allows optical subtraction of retinal circulation when focusing on the choroid and vice versa. Third, the instrument acquires and processes all data digitally, is personal computer-based, is compact, operates with a mouse-driven graphical user interface, and allows easy data exchange with conventional software. With further modifications in software and hardware, this device offers the possibility of producing a three dimensional map of the retinal and choroidal vasculature. PMID- 7485367 TI - Treatment of facial palsies with external eyelid weights. AB - PURPOSE: Because ocular exposure is a major complication of facial paralysis, an external gold eyelid load weight was evaluated for effectiveness in the treatment of this problem. METHODS: We prospectively studied 12 patients with unilateral facial paralysis who were treated with the external eyelid weights. Follow-up examinations included corneal exposure, amount of artificial tear usage, patient comfort, and complications associated with the weights. Treatment end points were the patient's decision to undergo a canthoplasty or placement of an implanted weight or resolution of the facial paresis to better than House's grade IV/VI. RESULTS: Of the 12 patients studied, ten had decreased corneal exposure on the affected side, with a coincident decrease in artificial tear drop use and increased comfort. One patient had no improvement in a corneal defect, and one was unable to apply the weight. Five patients had some difficulty in positioning the weight, which was related to upper eyelid dermatochalasis in four of the five. Only two weights were lost in over two years of total wearing time. CONCLUSIONS: External eyelid weights are useful in the treatment of ocular exposure associated with facial paralysis. The weights decreased corneal exposure, decreased reliance on artificial tear drops, and increased patient comfort. The weights were helpful as a trial before implantation of eyelid weights and as a longer-term treatment for ocular exposure in patients with temporary facial paralysis. Loose upper eyelid skin may limit their usefulness in some patients. PMID- 7485368 TI - Assessment of radial aspheres by the Arc-step algorithm as implemented by the Keratron keratoscope. AB - PURPOSE: To assess the accuracy with which the Keratron (Optikon 2000, Rome, Italy) measured rotationally symmetric, radially aspheric test surfaces according to an arc-step profile reconstruction algorithm and to discriminate between error caused by the algorithm and error from other sources. METHODS: Height, local power, and axial power calculated from radius of curvature centered on the instrument's axis were reported by the Keratron for four surfaces that had radial profiles similar to normal corneas. The Keratron profile reconstruction algorithm was simulated by using ray tracing. Keratron measurements were compared with the surfaces' formulas and the ray-traced simulations. RESULTS: The heights reported by the Keratron were within 0.25 microns from the four surfaces at less than 3 mm from the keratoscope axis and generally within 1 micron of the height calculated from the surfaces' formulas. The Keratron's axial powers were within +/- 0.1 diopter of the simulation of the axial solution between 1 and 4 mm of the axis but were greater central to 1 mm and peripheral to 4 mm. The Keratron's local powers were within -0.25 diopters at less than 4 mm from the axis and peripherally were between +1.75 diopters and -0.75 diopter of power calculated from the surface's instantaneous radii of curvature. Height error because of the arc-step algorithm was less than -0.2 micron. CONCLUSIONS: The Keratron's arc step profile reconstruction algorithm contributed to its ability to measure height more accurately than keratoscopes that use spherically biased algorithms and provided measurement of local power. PMID- 7485369 TI - Retinal nerve fiber layer. PMID- 7485370 TI - Erbium:YAG laser photoablation of human cornea. AB - PURPOSE: To determine whether the healing response of the human cornea after photoablation with a midinfrared solid-state laser was more pronounced than after excimer laser keratectomies because of the increased thermal damage in adjacent tissue. METHODS: We treated five blind human eyes with an erbium:YAG (Er:YAG) laser prototype and followed up the patients for nine months. RESULTS: Regarding epithelial healing and corneal clarity, the patients had a healing response very similar to that of excimer laser. Further clinical and experimental studies are necessary to estimate healing and regression. PMID- 7485371 TI - Vortex keratopathy associated with atovaquone. AB - PURPOSE: We studied a case of vortex keratopathy that was associated with the use of atovaquone. METHOD: A patient with acquired immunodeficiency syndrome (AIDS) with pulmonary Pneumocystis carinii was treated with 750 mg of atovaquone three times a day. RESULTS: Similar to previous findings of drug-induced vortex keratopathy, atovaquone vortex keratopathy is presumably caused by its lipophilic properties. This property is similar to that of other lipophilic agents such as amiodarone, chloroquine, chlorpromazine, quinacrine, and suramin. CONCLUSION: Atovaquone should be added to the list of agents causing vortex keratopathy involving the corneal epithelium. PMID- 7485372 TI - Interleukin-10 in the vitreous of patients with primary intraocular lymphoma. AB - PURPOSE: To find an accurate and simple method to diagnose primary intraocular lymphoma. METHODS: We prospectively analyzed concentrations of interleukin-10, interleukin-1, interleukin-2, interleukin-4, and interleukin-6 in vitrectomy specimens from three patients with primary intraocular lymphoma and five patients with uveitis by enzyme-linked immunosorbent assay. RESULTS: Interleukin-10 in the vitreous was detected in patients with primary intraocular lymphoma. Interleukin 10 levels correlated with clinical activity and the number of malignant cells. CONCLUSION: Concentrations of interleukin-10 in the vitreous may be a helpful clue to both clinicians and pathologists in diagnosing primary intraocular lymphoma. PMID- 7485373 TI - Rapidly progressive optic disk neovascularization after diabetic papillopathy. AB - PURPOSE: To determine if diabetic papillopathy may be associated with optic disk neovascularization and visual loss. METHODS: We examined two young adults with bilateral diabetic papillopathy who had permanent loss of vision associated with rapidly progressive posterior segment neovascularization. RESULTS: Within three months both patients developed florid optic disk neovascularization. This neovascularization was slow to regress despite full panretinal photocoagulation. One patient developed a macular tractional retinal detachment. CONCLUSIONS: Diabetic papillopathy may be associated with rapid progression of diabetic retinopathy and, in particular, optic disk neovascularization. Patients should be monitored for this possibility. PMID- 7485374 TI - Vogt-Koyanagi-Harada syndrome in a 4-year old child. AB - PURPOSE: We studied a case of severe bilateral Vogt-Koyanagi-Harada syndrome in a 4-year-old boy. METHODS: We evaluated the patient's clinical course. RESULTS: The patient had severe bilateral, nongranulomatous uveitis and mild uveitic glaucoma. Initial examination and laboratory evaluation failed to provide a diagnosis. The patient subsequently developed areas of vitiligo, alopecia, and poliosis, suggesting the diagnosis of Vogt-Koyanagi-Harada syndrome. This diagnosis was confirmed by the eventual development of bilateral neurosensory retinal detachments. Vision was lost despite aggressive therapy with corticosteroids and chlorambucil. CONCLUSION: Although uncommon, Vogt-Koyanagi-Harada may affect young children, and may be severe. PMID- 7485376 TI - A pigmented adenoma of the ciliary epithelium examined by magnetic resonance imaging. AB - PURPOSE: We studied a case of a pigmented adenoma of the ciliary epithelium. METHODS: We used magnetic resonance imaging in the clinical diagnosis of this tumor. RESULTS: The tumor was successfully treated by local excision. CONCLUSIONS: Magnetic resonance imaging is most useful in evaluating a ciliary body mass for local extension. It cannot clinically distinguish a pigmented adenoma of the ciliary epithelium from a uveal melanoma. PMID- 7485375 TI - Posterior scleritis with annular ciliochoroidal detachment. AB - PURPOSE: We studied an unusual case of posterior scleritis in a patient with sarcoidosis. METHOD: The medical record was reviewed for clinical manifestation, course, and tests performed, including laboratory evaluations, fluorescein angiography, and ultrasonography. RESULTS: The patient had posterior scleritis and unilateral angle-closure glaucoma caused by an annular ciliochoroidal detachment. Sarcoidosis was confirmed by biopsy of an enlarged parotid gland. CONCLUSION: The mechanism of angle-closure glaucoma may not be clinically or echographically apparent for a week or more in patients who develop annular ciliochoroidal detachment. We also found an unusual association of sarcoidosis and annular ciliochoroidal detachment secondary to posterior scleritis. PMID- 7485378 TI - Retinal photoreceptor dystrophies: LI. Edward Jackson Memorial Lecture. PMID- 7485377 TI - Bone formation in hydroxyapatite orbital implants. AB - PURPOSE: We examined the histopathologic features of two hydroxyapatite orbital implants that were removed from two patients. METHODS: Two surgically removed hydroxyapatite orbital implants were decalcified and processed for routine light microscopic examination. RESULTS: Both implants demonstrated fibrovascular ingrowth and foci of bone formation. CONCLUSION: Bone may form in hydroxyapatite orbital implants. PMID- 7485379 TI - Initial clinical experience with the Ahmed glaucoma valve implant--correction. PMID- 7485380 TI - gamma-Glutamyl transpeptidase. What does the organization and expression of a multipromoter gene tell us about its functions? AB - gamma-Glutamyl transpeptidase is a key enzyme in glutathione (GSH) salvage, metabolism of endogenous mediators such as leukotrienes and prostaglandins, detoxification of xenobiotics including environmentally important compounds and carcinogens, and cellular processes dependent on the oxidation/reduction of glutathione. The enzyme is widely distributed, and these functions often occur in separate tissues and in response to different stimuli. Evidence indicates that gamma-glutamyl transpeptidase plays a direct role in some hepatic and renal responses to injury. In the mouse gamma-glutamyl transpeptidase is a single copy gene expressed from at least seven promoters, and many of the transcribed gamma glutamyl transpeptidase RNAs are restricted in their expression. Studies that combine analyses of cellular processes with a knowledge of gene structure and expression hold promise for unravelling how these two different levels of function are integrated. PMID- 7485381 TI - Apolipoprotein A1-derived amyloid in human aortic atherosclerotic plaques. AB - Amyloid deposits in the aortic intima are very common in association with atherosclerosis and aging. In the present study, a major fibril protein purified from amyloid present in human atherosclerotic plaques was shown to be a 69-amino acid N-terminal fragment of apolipoprotein AI. Although senile form of localized apolipoprotein AI-derived amyloidosis has recently been documented in pulmonary vessels of dogs, this is the first example of a localized human amyloid derived from this apolipoprotein. PMID- 7485382 TI - Suppression of insulitis in non-obese diabetic (NOD) mice by oral insulin administration is associated with selective expression of interleukin-4 and -10, transforming growth factor-beta, and prostaglandin-E. AB - Oral administration of autoantigens suppresses development of autoimmunity in several animal models, and is being tested in clinical trials in patients with autoimmune diseases such as multiple sclerosis and rheumatoid arthritis. Non obese diabetic (NOD) mice spontaneously develop insulin-dependent diabetes mellitus at 15 to 20 weeks of age, after mononuclear cell (MNC) infiltration of the pancreatic islets of Langerhans and destruction of insulin-producing beta cells. We have previously shown that oral administration of insulin suppresses insulitis and development of diabetes in the NOD mouse. Oral insulin has no metabolic effect on blood glucose. Oral insulin mediates its effect through a T cell-dependent mechanism as shown by adoptive transfer and T cell depletion experiments, but the mechanisms responsible have not been fully explored. We now report a serial analysis of the cells and cytokines associated with development of diabetes in NOD mice, and contrast this with the findings in animals fed equine insulin or a control protein (ovalbumin). Animals were fed 1 mg twice a week for 5 weeks, beginning at 5 weeks of age. Marked insulitis in naive or ovalbumin-fed NOD mice occurred at 10 weeks, at which time a dense peri-islet and intra-islet MNC infiltration was observed. Immunohistological studies using monoclonal antibodies showed that infiltrating MNC consisted mainly of CD4+ T cells ( > 75% of leukocytes) plus smaller numbers of macrophages and CD8+ T cells. These cells displayed evidence of immune activation with expression of receptors for interleukin-2 (IL-2R) plus Th1 cytokines; dense labeling for IFN gamma and tumor necrosis factor-alpha, plus lesser amounts of IL-2, was observed. MNC lacked labeling for IL-4, IL-10, prostaglandin-E, or transforming growth factor-beta. By contrast, at 10 weeks, pancreatic tissues from NOD mice fed insulin showed considerably less insulitis, and the residual MNC, although still largely CD4+ T cells plus macrophages, showed dense labeling for IL-4, IL-10, prostaglandin-E, and transforming growth factor-beta and an absence of IL-2, IFN gamma or tumor necrosis factor-alpha Taken together with our previous findings, these data indicate that oral administration of insulin affects the development of diabetes in NOD mice through the generation of cells that elaborate immunoregulatory cytokines within the target organ and shift the balance from a Th1 to a Th2 pattern of cytokine expression. PMID- 7485383 TI - Long-term productive human immunodeficiency virus-1 infection in human infant microglia. AB - The course of human immunodeficiency virus 1 (HIV-1) infection in human infant microglia was studied using purified primary cultures of microglia derived from brain autopsy tissue. Previous in vitro studies have used fetal or adult brain tissue. Important differences may exist between brain tissues of different maturational ages with regard to HIV-1 replication and other neuropathogenic effects. Infant microglia were infected with four different strains of HIV-1 (JR FL, JR-CSF, Ba-L, and IIIB). Productive infection was demonstrated by p24 antigen production, immunocytochemistry, and recovery of replication-competent virus from the supernatants of the infected cultures. Multinucleated giant cells developed in culture mimicking the neuropathological changes seen in the brains of patients with HIV encephalopathy. Productive infection was more readily established by monocyte-tropic strains (JR-FL and Ba-L) of HIV-1 than by a lymphocyte-tropic strain (IIIB). p24 antigen production in this system peaked at 47 to 51 days postinfection. Viral persistence in giant cells was demonstrated by immunocytochemistry for the gp120 and gp41 viral antigens as late as 70 days postinfection. This in vitro culture system, using infant microglia that support viral replication for more than 2 months, may provide a useful model for studying the pathogenesis of progressive HIV encephalopathy. PMID- 7485385 TI - Local macrophage proliferation in multinucleated giant cell and granuloma formation in experimental Goodpasture's syndrome. AB - Granuloma is a specialized form of inflammatory reaction featuring focal macrophage and T-cell accumulation and multinucleated giant cell formation. It is widely held that macrophage accumulation within granulomatous lesions results from recruitment of blood monocytes, whereas proliferation of monocyte/macrophages makes little contribution to this process. The present study of macrophage proliferation within immunologically induced granulomas in rat experimental Goodpasture's syndrome challenges the conventional view. In this disease, granulomatous lesions in the kidney and lung contained 60 to 70% macrophages of an ED1+ED2-ED3-blood monocyte phenotype. However, double immunohistochemistry showed that up to 75% of ED1+ macrophages within granulomatous lesions were proliferating on the basis of proliferating cell nuclear antigen expression and bromodeoxyuridine incorporation. In contrast, no proliferating cell nuclear antigen expression or bromodeoxyuridine incorporations was detected in blood monocytes, indicating that proliferation of ED1+ED2-ED3- cells was a localized event within granulomatous lesions. A second finding of note was that almost all ( > 95%) nuclei within multinucleated giant cells were positive for proliferating cell nuclear antigen, but these nuclei lacked bromodeoxyuridine incorporation. This suggests a novel mechanism of multinucleated giant cell formation involving fusion of macrophages in G1 phase, which then halts progression into S phase of the cell cycle. In conclusion, this study has found that local macrophage proliferation plays an important role in the pathogenesis of granuloma formation. PMID- 7485384 TI - Expression of matrix proteinases during human intrahepatic bile duct development. A possible role in biliary cell migration. AB - Primitive biliary cells are known to migrate from the ductal plate into the mesenchyme during human intrahepatic bile duct development, and this migration process is essential for normal development of intrahepatic bile ducts. However, its molecular mechanism is unknown. Matrix proteinases play an important role in cell migration during cancer invasion and organ development. In this study, we therefore investigated in situ expression of matrix metalloproteinases (MMP) and tissue inhibitors of MMP (TIMP) during human intrahepatic bile duct development, using 32 human fetal livers. We also examined in situ expression of trypsinogen/trypsin, chymotrypsinogen/chymotrypsin, and cathepsin B, which are matrix proteinases and activators of MMP. MMP-1 expression was noted in the ductal plate and migrating primitive biliary cells. MMP-2, MMP-3, and MMP-9 were expressed in the ductal plate. TIMP-1 and TIMP-2 were expressed in the ductal plate and migrating primitive biliary cells. Trypsinogen/trypsin, chymotrypsinogen/chymotrypsin, and cathepsin B were also expressed in primitive biliary cells. These data suggest that MMP, trypsinogen/trypsin, chymotrypsinogen/chymotrypsin, and cathepsin B play a critical role in biliary cell migration during human intrahepatic bile duct development by degrading extracellular matrix proteins. The data also suggest that MMP inhibitors (TIMP-1 and TIMP-2) and MMP activators (trypsin, chymotrypsin, and cathepsin B) play an important role in biliary cell migration. The coordinated expression of MMP, MMP inhibitors, and MMP activators may be necessary for the normal development of human intrahepatic bile ducts. PMID- 7485386 TI - Chimeric TLS/FUS-CHOP gene expression and the heterogeneity of its junction in human myxoid and round cell liposarcoma. AB - Myxoid liposarcomas have a unique and specific t(12;16)q13;p11) chromosomal translocation. The breakpoint has recently been identified and shown to involve the TLS/FUS gene on chromosome 16 and the CHOP gene on chromosome 12. This translocation causes fusion of these genes resulting in the expression of a novel chimeric TLS/FUS-CHOP message. Using the polymerase chain reaction with primer sets derived from sequences of TLS/FUS and CHOP cDNAs, we could amplify three types of the fusion transcripts from seven of seven samples of myxoid and round cell liposarcomas. In six of the seven positive samples, two kinds of chimeric messenger RNAs were found that have been reported previously. However, the last sample had a novel chimeric message that had an extra sequence of 33 bp derived from the TLS/FUS gene. Thus, it was shown that these fusion transcripts had a varying extent of the sequence of TLS/FUS gene incorporated at the site of the fusion. However, the TLS/FUS-CHOP fusion transcripts were not detected in two pleomorphic liposarcomas or in three myxoid variants of malignant fibrous histiocytomas. Our findings indicate that in liposarcomas TLS/FUS-CHOP fusion transcripts have variations at the junction of chimeric messages, which was the case for Ewing's sarcoma. Detection of the chimeric message by reverse transcription polymerase chain reaction was also suggested to be a useful approach for the diagnosis of myxoid and round cell liposarcomas that have (12;16) translocation, and for distinguishing them from pleomorphic liposarcoma and myxoid variant of malignant fibrous histiocytomas. PMID- 7485387 TI - Immunohistochemical detection of active transforming growth factor-beta in situ using engineered tissue. AB - The biological activity of transforming growth factor-beta 1 (TGF-beta) is governed by dissociation from its latent complex. Immunohistochemical discrimination of active and latent TGF-beta could provide insight into TGF-beta activation in physiological and pathological processes. However, evaluation of immunoreactivity specificity in situ has been hindered by the lack of tissue in which TGF-beta status is known. To provide in situ analysis of antibodies to differentiate between these functional forms, we used xenografts of human tumor cells modified by transfection to overexpress latent TGF-beta or constitutively active TGF-beta. This comparison revealed that, whereas most antibodies did not differentiate between TGF-beta activation status, the immunoreactivity of some antibodies was activation dependent. Two widely used peptide antibodies to the amino-terminus of TGF-beta, LC(1-30) and CC(1-30) showed marked preferential immunoreactivity with active TGF-beta versus latent TGF-beta in cryosections. However, in formalin-fixed, paraffin-embedded tissue, discrimination of active TGF-beta by CC(1-30) was lost and immunoreactivity was distinctly extracellular, as previously reported for this antibody. Similar processing-dependent extracellular localization was found with a neutralizing antibody raised to recombinant TGF-beta. Antigen retrieval recovered cell-associated immunoreactivity of both antibodies. Two antibodies to peptides 78-109 showed mild to moderate preferential immunoreactivity with active TGF-beta only in paraffin sections. LC(1-30) was the only antibody tested that discriminated active from latent TGF-beta in both frozen and paraffin-embedded tissue. Thus, in situ discrimination of active versus latent TGF-beta depends on both the antibody and tissue preparation. We propose that tissues engineered to express a specific form of a given protein provide a physiological setting in which to evaluate antibody reactivity with specific functional forms of a protein. PMID- 7485388 TI - In situ mRNA hybridization technique for analysis of metastasis-related genes in human colon carcinoma cells. AB - The purpose of this study was to determine whether the expression level of several genes that regulate different steps of the metastatic process correlates with the metastatic potential of human colon carcinoma cells. The mRNA expression level for epidermal growth factor receptor (growth), basic fibroblast growth factor and interleukin-8 (angiogenesis), type IV collagenase (invasion), E cadherin and carcinoembryonic antigen (adhesion), and the multidrug resistance gene mdr-1 (drug resistance) in the human KM12 colon carcinoma cell lines and clones with different metastatic potential was measured by Northern blot analysis and by in situ hybridization technique. Highly metastatic KM12SM and KM1214 cells growing in culture uniformly expressed high levels of epidermal growth factor receptor, basic fibroblast growth factor, and carcinoembryonic antigen mRNA, whereas cultures of low metastatic KM12C, clone 1, clone 3, and clone 6 cells displayed heterogeneous patterns of expression. KM12C (low metastatic) and KM12SM (highly metastatic) cells were implanted into the subcutis (ectopic) or the wall of the cecum (orthotopic) of nude mice. The mRNA expression level for epidermal growth factor receptor, basic fibroblast growth factor, interleukin-8, type IV collagenase, carcinoembryonic antigen, and mdr-1 was increased in the cecal wall tumors as compared with subcutaneous tumors or in vitro cultures. These data demonstrate a direct correlation between constitutive and inducible expression of several metastasis-related genes and the metastatic potential of human colon carcinoma cells. PMID- 7485389 TI - Distributions of melanoma growth stimulatory activity of growth-regulated gene and the interleukin-8 receptor B in human wound repair. AB - The alpha-chemokines have been implicated as regulators of proliferation and differentiation of normal keratinocytes and as mediators of keratinocyte maturation and migration in inflammatory processes that involve the skin. Using the cutaneous wound repair model, we examined the sites and temporal sequence of the appearance of melanoma growth stimulatory activity or growth-regulated gene (MGSA/GRO;ligand) and the type B interleukin (IL)-8 receptor (IL-8RB) to which MGSA/GRO binds. Human burn tissues (n = 44) representing days 2 to 12 after injury were obtained during surgical debridement, fixed in 4% paraformaldehyde, and embedded in paraffin. Immunolocalizations were performed with polyclonal antisera for both ligand and receptor, as well as a monoclonal antibody for the IL-8 RB. Western blot analysis confirmed the presence of the IL-8 RB in immunoprecipitates of epidermal keratinocyte lysates. In normal skin, MGSA/GRO protein was restricted to sites populated by differentiated keratinocytes (suprabasal compartments, inner root sheath cells, and dermal sweat ducts). MGSA/GRO protein was barely detectable within epithelial margins and islands of burn wounds where the migrating/proliferating keratinocyte populations reside, but staining intensities increased as cells matured into the outer layers. Weak diffuse staining was detected in areas of neutrophilic infiltration (granulation tissue and overlying exudates). By contrast, in normal skin the IL-8 RB was detected in specific locations within epidermal and dermal compartments of healing wounds. In the dermis, polyvalent antibodies detected receptor immunoreactivity most prominently in dermal sweat ducts and endothelium of capillaries, whereas this immunoreactivity was inconspicuous in sections stained with the monoclonal antibody. Receptor immunostaining was noted in migrating/proliferating keratinocytes in epithelial margins and islands but was in the outer layers or in hypertrophic epidermis adjacent to wounds. This same pattern was observed in epidermal appendages such as hair follicles and eccrine sweat ducts. In granulation tissues, IL-8 RB was noted in numerous fibroblasts and in subpopulations of macrophages and smooth muscle. The presence of both MGSA/GRO and its receptor in human burn wounds implicate this cytokine as an autocrine or paracrine mediator of epidermal regeneration in both the inflammatory and proliferative phases of cutaneous wound repair. PMID- 7485390 TI - Growth factors in porcine full and partial thickness burn repair. Differing targets and effects of keratinocyte growth factor, platelet-derived growth factor BB, epidermal growth factor, and neu differentiation factor. AB - The topical application of recombinant growth factors such as epidermal growth factor, platelet-derived growth factor-BB homodimer (rPDGF-BB), keratinocyte growth factor (rKGF), and neu differentiation factor has resulted in significant acceleration of healing in several animal models of wound repair. In this study, we established highly reproducible and quantifiable full and deep partial thickness porcine burn models in which burns were escharectomized 4 or 5 days postburn and covered with an occlusive dressing to replicate the standard treatment in human burn patients. We then applied these growth factors to assess their efficacy on several parameters of wound repair: extracellular matrix and granulation tissue production, percent reepithelialization, and new epithelial area. In full thickness burns, only rPDGF-BB and the combination of rPDGF-BB and rKGF induced significant changes in burn repair. rPDGF-BB induced marked extracellular matrix and granulation tissue production (P = 0.013) such that the burn defect was filled within several days of escharectomy, but had no effect on new epithelial area or reepithelialization. The combination of rPDGF-BB and rKGF in full thickness burns resulted in a highly significant increase in extracellular matrix and granulation tissue area (P = 0.0009) and a significant increase in new epithelial area (P = 0.007), but had no effect on reepithelialization. In deep partial thickness burns, rKGF induced the most consistent changes. Daily application of rKGF induced a highly significant increase in new epithelial area (P < 0.0001) but induced only a modest increase in reepithelialization (83.7% rKGF-treated versus 70.2% control; P = 0.016) 12 days postburn. rKGF also doubled the number of fully reepithelialized burns (P = 0.02) at 13 days postburn, at least partially because of marked stimulation of both epidermal and follicular proliferation as assessed by proliferating cell nuclear antigen expression. In situ hybridization for KGFR in porcine burns revealed strong expression of KGFR on hair follicles and basal epidermis, confirming direct rKGF action on follicular as well as epidermal keratinocytes. Although the epithelial proliferation induced by rKGF resulted in marked neoepidermal psoriasiform hyperplasia with exaggerated rete ridges and neoepidermal and follicular maturation as assessed by expression of cytokeratin 10, a marker of keratinocyte terminal differentiation was not delayed and appeared to be accelerated in some rKGF-treated burns. Recombinant epidermal growth factor induced a trend toward increased new epithelial area in deep partial thickness burns, but had no effect on reepithelialization. The recombinant neu differentiation factor-alpha 2 isoform had no significant biological effects in either full or deep partial thickness burns.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7485391 TI - Apical topography and modulation of ICAM-1 expression on activated endothelium. AB - Leukocyte-endothelium interactions and general inflammatory responses are contributed by the regulated expression of intercellular adhesion molecule-1 (ICAM-1) on endothelium. It is now shown by confocal fluorescence microscopy and immunogold transmission electron microscopy that ICAM-1 was exclusively localized on the apical (luminal) membrane of cytokine-activated human umbilical vein endothelial cells. In contrast, other cell adhesion-promoting molecules, including beta 1 integrins, were expressed exclusively on the basolateral endothelial cell membrane, under the same experimental conditions. Kinetic binding studies of a 125I-labeled monoclonal antibody to ICAM-1 revealed that approximately 8% of membrane ICAM-1 on cytokine-activated endothelium was internalized in both coated and non-coated vesicles at 37 degrees C, with a t1/2 of approximately 18 min and a rate of approximately 3200 molecules/minute. This internalization pathway was directly dependent upon the level of ICAM-1 expression on the cell surface. Genetically engineered ICAM-1 transfectants, expressing a 10-fold higher receptor density than activated endothelium, internalized approximately 18% of membrane ICAM-1 at a rate of 75,000 molecules/minute with a t1/2 of approximately 22 min. These findings suggest that a combined pathway of polarized membrane topography and receptor trafficking may regulate ICAM-1-dependent adhesion at the site of vascular injury and endothelial cell activation. PMID- 7485392 TI - Blood-ocular barrier breakdown in eyes with ocular melanoma. A potential role for vascular endothelial growth factor/vascular permeability factor. AB - A series of 130 eyes with ocular melanomas, 19 normal eyes, and 18 eyes affected with other disorders leading to blood-ocular barrier (BOB) breakdown were immunohistochemically stained for albumin to localize sites of BOB failure within the retina, ciliary body, and iris. Thirty-nine of the eyes containing melanomas and all of the other eyes were also immunohistochemically stained for vascular endothelial growth factor (VEGF), to investigate its potential role as a mediator for BOB failure. Eyes with melanomas showed widespread leakage through the retinal pigment epithelium, and 58% demonstrated leakage from retinal vessels in the proximity of the tumor. BOB failure remote from the tumor also occurred in retina (50%), optic nerve head (77%), ciliary body (51%), and iris (51%), suggesting that a soluble mediator may be involved. VEGF was demonstrated intraretinally in the proximity of (46%) and remote from (24%) melanomas and in eyes affected by other disease processes, particularly those involving neoplasia or retinal detachments, usually within particular cell populations (ie, retinal vessel walls, ganglion cells, inner or outer nuclear layers, retinal pigment epithelium). VEGF localization in retina, ciliary body, and iris often coincided with sites of extravasated albumin. Preincubation of albumin or VEGF antibodies with normal serum or VEGF peptide, respectively, eliminated or markedly reduced all immunoreactivity. Only 1 of 14 normal postmortem eyes and 0 of 5 normal surgically removed eyes showed VEGF positivity in the retina, 5 of 19 normal eyes had weak positivity in the ciliary body, and VEGF was not demonstrated in the iris of normal eyes. VEGF cannot account for all of the BOB failure associated with ocular melanomas, but appears likely to play a contributing role in many cases. PMID- 7485394 TI - Apolipoprotein AI-derived pulmonary vascular amyloid in aged dogs. AB - Our studies confirm the common occurrence of a unique form of apolipoprotein AI (apoAI)-derived vascular amyloidosis in dogs that appears to be unrelated to other disease conditions, but is associated with aging. Vascular amyloid deposits were most frequently located within the intima and media of medium-sized pulmonary arteries, and were not confirmed in any other tissues. Pulmonary vascular amyloid immunoreactive with antiserum to purified N-terminal (1-71) canine apoAI amyloid protein was demonstrated retrospectively in 12.8% of necropsied dogs (N = 243) 10 years of age or older. In a subsequent expanded 1 year prospective study of necropsied dogs (N = 231) of all ages, apoAI-derived pulmonary vascular amyloid deposits were demonstrated in 0.7% of dogs < 10 years of age and in 22% of dogs 10 years of age or older. The incidence of this form of amyloid in dogs 10 years of age or older was significantly associated with advancing age (P < 0.00001). However, significant differences regarding gender, breed, or the frequency of selected common disease conditions were not observed when the dogs with apoAI-derived amyloid were compared with control dogs. The possibility that this new form of senile apoAI-derived amyloidosis is a manifestation of an age-associated aberration in apoAI metabolism or is related to a mutant form of apoAI is the subject of ongoing investigations. PMID- 7485393 TI - Expression of c-ets-1, collagenase 1, and urokinase-type plasminogen activator genes in lung carcinomas. AB - The c-ets-1 transcription factor has been involved in the in vitro transactivation of matrix-degrading protease genes that might play an important role in tumor invasion. Using in situ hybridization, we analyzed serial frozen sections for c-ets-1, collagenase 1, and urokinase-type plasminogen activator gene expression in 54 lung carcinomas including 34 non-neuroendocrine carcinomas (18 squamous carcinomas, 10 adenocarcinomas, 3 large cell carcinomas, and 3 basaloids) and 20 neuroendocrine carcinomas (7 small cell lung carcinomas, 4 large cell neuroendocrine carcinomas, 4 well differentiated neuroendocrine carcinomas, and 5 carcinoids). c-ets-1 gene was expressed in stromal cells in 44/54 lung carcinomas including one metastasizing carcinoid. c-ets-1 transcripts were also detected in cancer cells more frequently in neuroendocrine than in non neuroendocrine carcinomas (P = 0.0059) and in stages III and IV and metastasis more frequently than in stages I and II ( P = 0.0065). Collagenase 1 gene was expressed in 16/34 non-neuroendocrine tumors and in 1/20 neuroendocrine tumors, either in stromal (12/17) or in cancer cells (6/17). Urokinase-type plasminogen activator mRNAs were expressed in 45/54 lung carcinomas in stromal and/or cancer cells. In non-neuroendocrine tumors, c-ets-1 and collagenase 1 gene expressions in stromal cells were correlated. These results demonstrate that the transcription factor c-ets-1, collagenase 1, and urokinase-type plasminogen activator are involved in lung cancer invasion and suggest that c-ets-1 protein might transactivate collagenase 1 gene during tumor invasion. PMID- 7485395 TI - Regulation of fibrin deposition by malignant mesothelioma. AB - Malignant mesothelioma (MM) is a locally aggressive tumor that spreads by poorly understood mechanisms. Because neoplastic spread has been linked to altered fibrin turnover, we used immunohistochemistry of nine MM and three fibrous tumors of the pleura to confirm in vivo fibrin deposition and expression of selected coagulation and fibrinolytic reactants in MM. Tumor-associated fibrin was readily detectable at site of tissue invasion. Little fibrin was distributed within the tumor, but tissue factor and tissue factor pathway inhibitor, urokinase, urokinase receptor, and plasminogen activator inhibitors 1 and 2 were all detected in either epithelioid or sarcomatous areas of MM. We used the MS-1 human pleural mesothelioma cell line to determine how expression of these reactants is regulated. Fibrinolytic activity of MS-1 is mainly due to urokinase and is responsive to cytokine stimulation. Functional extrinsic activation and prothrombinase complexes assemble at the cell surface. MM express procoagulants as well as fibrinolytic reactants in vivo and in vitro that promote local fibrin formation and remodeling. Fibrin deposition occurs primarily at areas of tissue invasion and could promote local extension of this neoplasm. Sparsity of fibrin within the central portions of the tumor stroma suggests that local resorption of transitional fibrin occurs at sites of established MM. PMID- 7485396 TI - Chronic systemic treatment with epidermal growth factor in pigs causes pronounced urothelial growth with accumulation of glycoconjugates. AB - Epidermal growth factor (EGF) is present in large amounts in the urine, but the effects of systemically administered EGF on the urinary tract have not been described previously. In the present paper, we describe a potent growth induction of EGF on the urinary tract. Goettingen minipigs were treated with solvent (n = 5), EGF 30 micrograms/kg/day (n = 6) for 4 weeks, or EGF 30 micrograms/kg/day for 5 weeks followed by 3 weeks of recovery (n = 5). The ureters and bladders were examined by routine histology and electron microscopy and were immunostained for proliferating cell nuclear antigen. Four weeks of EGF treatment increased the median cross sectional area of the ureter fourfold with growth of all wall layers. The urothelium was widened from 5 cell layers in the controls to 10 in the EGF-treated animals. Proliferating cell nuclear antigen immunostaining revealed an increased mitotic activity in the basal zone of the urothelium. In the luminal zone, glycoconjugates accumulated in goblet cells, in cells with intracytoplasmic lumina, and beneath the luminal cell membrane in the umbrella cells. Our studies present a new experimental approach to growth induction of the urinary tract. The findings implicate the EGF system in regulating urothelial growth and glycoconjugate biosynthesis. PMID- 7485397 TI - Neurofibromatosis type 2 protein co-localizes with elements of the cytoskeleton. AB - The product of the neurofibromatosis type 2 (NF2) tumor suppressor gene is a 595 amino-acid protein bearing resemblance to a family of band-4.1-related proteins. These proteins, including ezrin, radixin, and moesin, probably function as molecular linking proteins, connecting the cytoskeleton to the cell membrane. On the grounds of the homology to the ezrin, radixin, and moesin proteins and on the basis of its predicted secondary structure, the NF2 protein is also thought to act as a cytoskeleton-cell membrane linking protein. Using monoclonal antibodies to amino- and carboxyl-terminal synthetic NF2 peptides we demonstrate the co localization of the NF2 protein with elements of the cytoskeleton in a COS cell model system and in cultured human cells. Furthermore, the presence of the NF2 protein in tissue sections is shown. The monoclonal antibodies specifically stain smooth muscle cells and the stratum granulosum of the human epidermis. In cultured smooth muscle cells the NF2 protein co-localizes with actin stress fibers. Immunoelectron microscopy demonstrates the presence of the NF2 protein associated with keratohyalin granules and to a lesser extent with intermediate filaments in the human epidermis. We conclude that the NF2 protein is indeed associated with multiple elements of the cytoskeleton. PMID- 7485398 TI - Phenotypic analysis of pulmonary perivascular mononuclear infiltrates that occur as a direct result of acute lethal graft-versus-host disease describes the onset of interstitial pneumonitis. AB - We recently determined that the sequential development of interstitial pneumonitis and lymphocytic bronchiolitis/bronchitis occurs as a direct result of acute lethal graft-versus-host disease. Interstitial pneumonitis develops before lymphocytic bronchiolitis/bronchitis primarily from the dissemination of perivascular mononuclear infiltrates. We have used the adult, nonirradiated (DA x LEW) F1 hybrid rat in the absence of chemotherapy, immunosuppression, or overt infection to determine the phenotype of infiltrating perivascular mononuclear cells throughout acute lethal graft-versus-host disease. F1 animals were intravenously injected with 1 x 10(6) DA parental lymphoid cells/g body weight, which produced 100% morbidity and mortality by day 21. Graft-versus-host disease animals were killed on days 3, 7, 10, 14, and 15 to 21 after injection. Whole left lung lobes were frozen, serially sectioned (4 microns), and incubated with a panel of mouse anti-rat monoclonal antibodies. Labeled antibody density was determined by computerized image analysis. Perivascular infiltration was observed first for ED1+, OX8+, and W3/25+ cells, and then OX41+, W3/13+ and OX19/25+ populations. OX6 was expressed in control tissues and at all time points tested. OX12+, OX39+ and MOM/3F12/F2+ cells were not quantifiable. The present study has determined that the process of perivascular infiltration was produced through a biphasic influx of OX6+, T-cell, and macrophage populations. PMID- 7485399 TI - Expression of alternatively spliced fibronectin variants during remodeling in proliferative glomerulonephritis. AB - Fibronectin (Fn) plays an important role in tissue remodeling during embryogenesis, wound repair, and vascular disease, and is thought to regulate cellular processes such as cell adhesion, migration, proliferation, and differentiation through specialized domains within the molecule. In addition, Fn can be alternatively spliced at three regions: extradomains EIIIA, EIIIB, and a variable segment V, potentially giving rise to functionally distinct variants of the molecule. We have previously shown a sequential expression of cellular Fn first by platelets, followed by macrophages, then mesangial cells in habu snake venom-induced proliferative glomerulonephritis (Am J Pathol 145: 585-597, 1994). These studies examined the cellular sources and glomerular localization of Fn in general but did not distinguish between the various alternatively spliced isoforms. In this study, we examine by in situ hybridization and immunohistochemistry the temporal expression and cellular sources of EIIIA, EIIIB, and V in a model of proliferation glomerulonephritis that has cell migration, proliferation, and extracellular matrix synthesis as features of tissue remodeling. Macrophages were the first cells to express Fn mRNA showing an EIIIA+, EIIIB-, and V95+ pattern beginning at 8 hours after habu snake venom injection. Migrating mesangial cells at the margins of early lesions (8 and 24 hours) did not overexpress mRNA encoding these Fn variants, but immunofluorescence microscopy revealed V95 and EIIIA protein at the margins of lesions. EIIIB was absent in lesions at this time. At 48 hours and peaking at 72 hours after habu snake venom injection, mesangial cells in central aspects of glomerular lesions expressed abundant mRNA and protein for V95 and EIIIA. EIIIB mRNA and protein was slight in the mesangium at these times. Parietal epithelial cells, particularly adjacent to glomerular lesions, also expressed abundant mRNA and protein for all three variants throughout the course of the disease, beginning at 24 hours after habu snake venom injection. Expression of mRNA and protein for all three isoforms declined by 2 weeks after habu snake venom injection. These studies show that migrating mesangial cells do not require their own synthesis of Fn and suggest that they might rely on exogenous sources of Fn, particularly V95+ and EIIIA+ forms. Commencement of enhanced expression of EIIIA and EIIIB mRNA and protein by resident glomerular cells coincided with the temporal course of cell proliferation, acquisition of alpha-smooth muscle cell actin phenotype, and matrix synthesis, suggesting that Fn isoforms have specific functions during the course of glomerular remodeling. PMID- 7485400 TI - Autocrine growth regulation of human glomerular mesangial cells is primarily mediated by basic fibroblast growth factor. AB - For various forms of human glomerulonephritis a close relationship between inflammatory injury and a local mesangial proliferative response has been described. Herein, we used primary cultures of human glomerular mesangial cells (HMCs) from five different donors to determine the autocrine growth-inducing capacity of their supernatants after stimulation with different cytokines and lipopolysaccharide (LPS) to determine whether this effect is due to basic fibroblast growth factor (bFGF). The basal growth-inducing capacity of supernatants collected from serum-free cultured HMC and concentrated 100-fold above a cut-off size of 10 kd was significantly increased by interleukin (IL)-1 beta, platelet-derived growth factor (PDGF), and LPS up to 15-fold, but not by IL 1 alpha, IL-6, or bFGF. An anti-human bFGF antibody blocked the majority of IL-1 or LPS-induced proliferative effect of supernatants; complete inhibition was achieved by a combination of anti-human bFGF- and anti-human platelet-derived growth factor antibodies. HMCs express different isoforms of bFGF (18, 21.5, and 24 kd) in membrane, cytosolic, and nuclear fractions. All isoforms of bFGF were found in the nuclear fraction of HMC, whether stimulated or not. Immunoblots for bFGF protein of HMC demonstrated that only a approximate to 16 kd bFGF protein was released into HMC supernatants after stimulation with IL-1 beta, platelet derived growth factor-BB, and LPS. The 18 kd isoform of bFGF accumulated in the membranes but was not released after stimulation with IL-1 alpha, IL-6, and bFGF, suggesting that its release was a prerequisite for autocrine growth stimulation. By means of reverse transcription polymerase chain reaction controlled by Southern blots, bFGF-mRNA expression of HMC was enhanced by IL-1 alpha, IL-1 beta, and LPS. Finally, we were able to show that HMCs are expressing bFGF receptors. In summary, our data demonstrate for the first time that the autocrine proliferative response of HMC to major inflammatory factors may primarily be mediated by bFGF. PMID- 7485402 TI - Burkitt's lymphomas express VH genes with a moderate number of antigen-selected somatic mutations. AB - The normal counterpart of the neoplastic B cells occurring in Burkitt's lymphomas (BL) is an issue of controversial debate. To clarify this matter, a semi-nested primer polymerase chain reaction was performed to amplify the VDJ rearrangements of the immunoglobulin heavy chain (VH) gene of DNA extracts from 10 (8 sporadic and 2 endemic) BL cases. The resulting amplificates were sequenced for comparison with known germ line VH segments. The control cases comprised six cases of B cell chronic lymphocytic leukemia and six cases of mantle cell lymphoma known to display naive nonmutated, ie, pre-germinal center VH configurations; and eight cases of follicular center lymphoma known to display mutated VH genes with signs of a still-ongoing mutation reaction, characteristic for germinal center cells and lymphomas that derive therefrom. The results of this approach revealed that both sporadic and endemic BL express mutated VH genes with a mutation frequency considerably lower (4.9% and 5.4%, respectively) than that observed in follicular center lymphoma (11.8%). In addition, after subcloning the amplificates, sequence analysis revealed no signs of ongoing mutations. These results led us to conclude that the derivation of neoplastic B cells in BL is definitely not from naive, nonmutated pre-germinal center B cells. Instead, our findings support the view that BL cells stem either from early centroblasts that are arrested after an initial hypermutation reaction, or from germinal center B cells that have differentiated in terms of surface immunoglobulin profile and mutation pattern but not in terms of morphology and proliferation toward SIgM+ IgD- memory B cells because of the deregulated c-myc gene expression. PMID- 7485401 TI - Uroplakins, specific membrane proteins of urothelial umbrella cells, as histological markers of metastatic transitional cell carcinomas. AB - Uroplakins (UPs) Ia, Ib, II, and III, transmembrane proteins constituting the asymmetrical unit membrane of urothelial umbrella cells, are the first specific urothelial differentiation markers described. We investigated the presence and localization patterns of UPs in various human carcinomas by applying immunohistochemistry (avidin-biotin-peroxidase complex method), using rabbit antibodies against UPs II and III, to paraffin sections. Positive reactions for UP III (sometimes very focal) were noted in 14 of the 16 papillary noninvasive transitional cell carcinomas (TCCs) (88%), 29 of the 55 invasive TCCs (53%), and 23 of the 35 TCC metastases (66%). Different localization patterns of UPs could be distinguished, including superficial membrane staining like that found in normal umbrella cells (in papillary carcinoma), luminal (microluminal) membrane staining (in papillary and invasive carcinoma), and, against expectations, peripheral membrane staining (in invasive carcinoma). Non-TCC carcinomas of various origins (n = 177) were consistently negative for UPs. The presence of UPs in metastatic TCCs represents a prime example of even advanced tumor progression being compatible with the (focal) expression of highly specialized differentiation repertoires. Although of only medium-grade sensitivity, UPs do seem to be highly specific urothelial lineage markers, thus operating up interesting histodiagnostic possibilities in cases of carcinoma metastases of uncertain origin. PMID- 7485403 TI - Detection of Epstein-Barr virus in multiple sites involved by Hodgkin's disease. AB - Tissues obtained from 14 patients with multiple anatomic sites involved by Hodgkin's disease were studied for Epstein-Barr virus (EBV) using in situ hybridization for EBV-encoded RNA (EBER) 1 and immunohistochemical methods for EBV latent membrane protein (LMP) expression. Each patient in this study had two to five separately involved anatomic sites, and all biopsy sites, a total of 43 specimens, were analyzed for EBV. EBV was detected in 6 of 14 (42.8%) patients with Hodgkin's disease, including 5 of 11 (45.4%) with nodular sclerosis and 1 of 3 (33%) with mixed cellularity. In these six patients, all biopsy sites were positive for both EBER1 and LMP. In the EBV-positive cases were analyzed the 3' end of the EBV LMP1 gene in al sites of disease using polymerase chain reaction. In three patients all sites of disease had a 30-base pair deletion. In two patients, there was discordance between sites of disease, with LMP1 gene deletions in some sites and other sites with the LMP1 gene in the germline configuration. The results of this study demonstrate that EBV, when found in Hodgkin's disease, is detectable in all anatomic sites involved. The presence of the same 30-base pair deletion in the EBV LMP1 gene in all sites of disease in three patients suggests that the deletion occurred before dissemination and that all sites are clonally related. However, the discordance between anatomic sites in two patients suggests that LMP1 gene deletion may also occur as a later event, after dissemination. These results lend further support to the hypothesis that EBV plays a role in the pathogenesis of a subset of cases of Hodgkin's disease. PMID- 7485404 TI - Crypt fission in the small intestine and colon. A mechanism for the emergence of G6PD locus-mutated crypts after treatment with mutagens. AB - In the small intestine and colon, administration of mutagens leads to the emergence of crypts populated by cells with a different, mutated phenotype. This is preceded by a transient rise in the frequency of crypts with a partially mutated phenotype, and the disappearance of these partially mutated crypts occurs contemporaneously with the attainment of a plateau value of the wholly mutated crypts. Here, using the mutagen ethyl nitrosourea and loss of glucose-6-phosphate dehydrogenase staining as a marker, we show that the plateau is reached at between 4.6 and 7 weeks in the colon and at 12 weeks in the small intestine of the same mice. Explanations for this difference have included differences in the stem cell cycle time of a single "master" stem cell or multiple stem cells occupying a stem cell "niche" with random loss after stem cell division. However, we demonstrate that the crypt fission index, or the incidence of crypts in fission, is some four times higher in the colon than in the small intestine at the time of ethyl nitrosourea injection, and propose an alternative hypothesis based on crypt fission as the mechanism for the more rapid evolution of wholly mutated crypts in the colon. The hypothesis should enable us to predict the results of future experiments, namely that the emergence of wholly mutated crypts is proportional to the crypt fission index. PMID- 7485406 TI - Pathology and immunohistochemistry of callitrichid hepatitis, an emerging disease of captive New World primates caused by lymphocytic choriomeningitis virus. AB - Callitrichid hepatitis is an arenavirus infection that recently emerged as a highly fatal disease of New World primates in the Callitrichidae family. As we previously reported, these primates develop hepatitis after contact with mice that are infected with variants of LCMV (LVMCCH), recently determined to have 86% identity with GC-P gene of the Armstrong and Western strains of LCMV. Here, we describe the histopathological lesions and tissue localization of viral antigens in confirmed cases of callitrichid hepatitis from recent outbreaks in two U.S. zoos. The liver in marmosets and tamarins with fatal infections consistently showed degeneration, necrosis, and inflammation, with variable involvement of the spleen, lymph nodes, adrenal glands, intestine, pancreas, and central nervous system. Lymphocytic choriomeningitis virus antigens were identified immunohistochemically in necrotic foci in these organs as well as in nondegenerating areas in lungs, kidney, urinary bladder, brain, and testes. The multi-organ tropism and histological pattern of LCMV infection in marmosets and tamarins are similar to those reported for the highly virulent arenavirus that causes Lassa fever in humans. Comparative studies of callitrichid hepatitis and Lassa fever would therefore be mutually beneficial for human and nonhuman primate medicine. PMID- 7485405 TI - Liver failure induces a systemic inflammatory response. Prevention by recombinant N-terminal bactericidal/permeability-increasing protein. AB - The observed increased susceptibility of patients with fulminant hepatic failure for local and systemic infections has been hypothesized to be due to a failure for the hepatic clearance function and subsequent leaking of endogenous endotoxins into the systemic circulation. However, experimental evidence for such a systemic inflammation during liver failure due to endogenous endotoxemia is lacking. Therefore, we designed a study to clarify whether circulating endotoxins due to liver failure could lead to the development of systemic inflammations. In a rat model for liver failure induced by a two-thirds partial hepatectomy, we evaluated the course of circulating tumor necrosis factor and interleukin-6, changes in blood chemistry and hemodynamics, and histopathological changes in the lungs. Partially hepatectomized animals, but not sham-operated animals, demonstrated cardiac failure, increased levels of creatinin and urea, metabolic acidosis, high plasma levels of tumor necrosis factor and interleukin-6, and an influx of PMNs in the lungs-together indicating the development of a systemic inflammatory response. Continuous infusion of recombinant N-terminal bactericidal/permeability-increasing protein (rBPI23), a well described endotoxin neutralizing protein, prevented these inflammatory reactions. Ex vivo experiments with rat plasma samples confirmed the presence of circulating endotoxins in partially hepatectomized rats as opposed to those treated with rBPI23. Thus, our results indicate that the early phase of liver failure induces a systemic inflammatory response triggered by circulating endotoxins, which can be prevented by perioperative infusion of rBPI23. PMID- 7485407 TI - NF2 gene analysis distinguishes hemangiopericytoma from meningioma. AB - The histogenesis of dural-based or "central" hemangiopericytomas (cHPCs) remains controversial. Some authors consider these tumors variants of meningiomas while others consider them akin to peripheral hemangiopericytomas (pHPCs). Meningiomas frequently have mutations in the neurofibromatosis 2 (NF2) gene, providing a molecular marker for meningiomas and other NF2-related tumors. We therefore analyzed the NF2 gene in cHPCs, pHPCs, and meningiomas to determine whether cHPCs are more similar at the molecular genetic level to meningiomas or pHPCs. Using paraffin-embedded archival material from 28 cHPCs (including three primary and recurrent tumors), 10 pHPCs, and 26 meningiomas, we scanned all 17 exons of the NF2 gene and flanking intronic sequences for mutations with single strand conformation polymorphism analysis and DNA sequencing. No NF2 mutations were found in either cHPCs or pHPCs, whereas 35% of meningiomas had NF2 gene alterations (P < 0.001). The NF2 gene mutations in meningiomas were all truncating mutations, consistent with previous studies. Our findings suggest that cHPCs are distinct from meningiomas at the molecular genetic level and support prior clinico-pathological data that distinguish these tumor-entities. PMID- 7485408 TI - The role of pancreatic islets in experimental pancreatic carcinogenicity. AB - Our previous studies have suggested that the presence of intact islets is essential for the induction of pancreatic exocrine tumors in the Syrian hamster model. To validate this, we investigated the effect of the carcinogen, N nitrosobis(2-oxo-propyl)amine (BOP) in hamsters, in which homologous isolated intact islets were transplanted into the submandibular gland (SMG). Freshly isolated pure islets from hamster donors were transplanted into the left SMG of 20 female host hamsters. Ten of these hamsters (group 1) received BOP (40 mg/kg) weekly for 3 weeks. Another 10 hamsters (group 2) were kept untreated. In groups 3 and 4 (10 hamsters each) the salt solution or isolated pancreatic ductal cells, respectively, was injected into the gland. In other groups (10 hamsters each) islets were transplanted into the peri-SMG connective tissue (group 5) or into the renal subcapsular space (group 6). Hamsters of group 1 (40 mg/kg, weekly for 3 weeks) as were group 7 hamsters, which served as BOP-treated controls. All BOP treated hamsters developed pancreatic lesions. Similar hyperplastic and atypical ductal/ductular proliferation and in situ carcinoma were found in the SMG of many group 1 hamsters. No such lesions were found in the SMG, peri-SMG, or renal subcapsular space of the other groups. Islets appear to be involved in carcinogenicity of BOP. The mechanism is obscure. PMID- 7485409 TI - Cortical and brainstem-type Lewy bodies are immunoreactive for the cyclin dependent kinase 5. AB - The immunoreactivity of cortical and brainstem-type Lewy bodies has been investigates with antibodies to the cyclin-dependent kinase 5 (cdk5), to the extracellular regulated kinase 1 (ERK-1), and to the cdc2p34 kinase and with antibodies specific for phosphorylation epitopes typical of paired helical filament-tau (PHF-tau). Both cortical and brainstem-type Lewy bodies in diffuse Lewy body disease and brainstem-type Lewy bodies in Parkinson's disease were found to be immunoreactive for cdk5 but not for cdc2p34 or ERK-1 or with the PHF tau antibodies. Double immunolabeling showed that cdk5-positive Lewy bodies were also ubiquitin immunoreactive and that cdk5 antibodies labeled as many Lewy bodies as ubiquitin antibodies in adequately fixed tissue. The cdk5 immunoreactivity of Lewy bodies was abolished by preabsorption of the antibody with a cdk5 peptide. The antibodies to cdk5 labeled a single 33-kd species on Western blots of human brain homogenates, with a similar intensity in control, diffuse Lewy body disease, and Alzheimer's disease, and this cdk5 species was found mainly in the particulate fraction of brain homogenates. This observation suggests that cdk5 might be a protein kinase involved in the phosphorylation of a molecular component of Lewy bodies, for example, neurofilament proteins known to be present in these inclusions. PMID- 7485410 TI - Interleukin-1 receptor antagonist decreases the number of necrotic neurons in rats with middle cerebral artery occlusion. AB - Marked increases in the brain expression of interleukin (IL)-1 have been reported in rats after permanent occlusion of a large cerebral artery. Interactions between endothelial cells and leukocytes have been implicated in the pathogenesis of several types of ischemic injury to the myocardium and other organs. In this study we asked whether inhibiting the effects of IL-1 would affect the outcome of an experimental brain infarct. Adult male Wistar rats (n = 13) with permanent occlusion of the middle cerebral artery were given IL-1 receptor antagonist. A second group (n = 13) with the same type of brain injury was given a placebo. A third group, subjected to a sham operation, was given either IL-1 receptor antagonist (n = 2) or a placebo (n = 2). Experiments were terminated after either 24 hours or 7 days. Compared with the control group, animals treated with IL-1 receptor antagonist improved their neurological score (P < 0.05), experienced less pronounced changes in body weight (P < 0.05), and had fewer necrotic neurons (P < 0.001) and fewer leukocytes in the ischemic hemisphere (P < 0.001) as well as a smaller area of pallor (P < 0.05) in the ischemis hemisphere. The results suggest that inhibiting the proinflammatory effects of IL-1 with a receptor antagonist is an effective way of influencing the leukocyte responses elicited by an arterial occlusion. Such leukocyte inhibition seemingly attenuates the number of necrotic neurons resulting from the occlusion of a large brain artery. PMID- 7485412 TI - Distribution of type XV collagen transcripts in human tissue and their production by muscle cells and fibroblasts. AB - Type XV collagen is a recently identified member of the diverse family of collagens, its structure being characterized by extensive interruptions in the collagenous sequences. A combination of Northern blot hybridization of fetal and adult human tissues and in situ hybridization analyses of a fetus with Down's syndrome, several placentas, and adult skin were used to localize expression of its mRNAs. Northern blot analysis revealed marked expression in heart, skeletal muscle, and placenta tissues and moderate levels in the kidney and pancreas. Clear in situ hybridization signals were detected in fibroblasts and endothelial cells in all tissues studied. Examination of fetal heart, skeletal muscle, and smooth muscle tissues showed that the high type XV collagen mRNA level in the muscle RNA was localized not only to fibroblasts residing in the endomysium but also to myoblasts. Interestingly, type XV collagen mRNAs were also synthesized by certain epithelial cells in kidney, lung, pancreas, and placenta. It was the morphologically immature glomeruli in the kidney and the lower parts of the nephron, especially the collecting ducts, that contained these mRNAs but not the mature glomeruli or proximal tubules, suggesting differences in expression during development. These findings indicate a wide distribution of type XV collagen transcripts, the main producers being mesenchymally derived cells, particularly muscle cells and fibroblasts. PMID- 7485413 TI - Glomerular inflammation in pregnant rats after infusion of low dose endotoxin. An immunohistological study in experimental pre-eclampsia. AB - Increased endotoxin sensitivity during pregnancy occurs in many animals, including rats. The mechanism of this phenomenon is not understood. In the present study it was investigated whether this increased sensitivity is reflected by an altered inflammatory pattern. Inflammatory cell influx, the O2(-)-producing potential of these cells, and expression of adhesion molecules was studied in the glomeruli of pregnant and cyclic rats at various intervals after low dose endotoxin infusion. Kidney sections were stained for monocytes and adhesion molecules (ICAM-1, VCAM-1, LFA-1, and VLA-4) using monoclonals, while potentially O2(-)-producing neutrophils (ie, activated neutrophils) were quantified using immunohistochemical methods. The results show early glomerular influx of activated neutrophils, maximally 4 hours after endotoxin. Both absolute neutrophil counts and relative numbers of activated neutrophils were significantly increased in pregnant versus cyclic rats. In contrast to cyclic rats, showing transient monocyte influx, in pregnant endotoxin-treated rats monocyte influx reaches a maximum at t = 168 hours. These cell kinetics were paralleled by expression of the various adhesion molecules. It was concluded that pregnancy profoundly influences not only the inflammation kinetics after endotoxin, but also the violence of the reaction, reflected by activated neutrophils. This altered glomerular inflammatory pattern may help to explain why low dose endotoxin infusion induces pre-eclamptic-like symptoms (such as an intraglomerular prothrombotic microenvironment and proteinuria) exclusively in the pregnant rat. PMID- 7485414 TI - Lessons to be learned from hematopoietic malignancies. Workshop report from the Division of Research Grants, National Institutes of Health. PMID- 7485411 TI - Interleukin-4 induces foreign body giant cells from human monocytes/macrophages. Differential lymphokine regulation of macrophage fusion leads to morphological variants of multinucleated giant cells. AB - Interleukin-4 induced the formation of foreign body-type giant multinucleated cells from human monocyte-derived macrophages, an effect that was optimized with either granulocyte-macrophage colony-stimulating factor or interleukin-3, dependent on the concentration of interleukin-4, and specifically prevented by anti-interleukin-4. Very large foreign body giant cells and, predominantly, giant cell syncytia with randomly arranged nuclei and extensive cytoplasmic spreading (285 +/- 121 nuclei and 1.151 +/- 0.303 mm2 per syncytium) were consistently obtained. Under otherwise identical culture conditions, relatively much smaller Langhans-type giant cells with circularly arranged nuclei were induced with a previously described combination of interferon-gamma plus granulocyte-macrophage colony-stimulating factor or interleukin-3 (16 +/- 6 nuclei and 0.033 +/- 0.013 mm2 per giant cell); their formation was prevented by anti-interferon-gamma but not by anti-interleukin-4. Similar rates of macrophage fusion were obtained in both culture systems (72 +/- 5% and 74 +/- 6%, respectively), but these two morphological variants did not occur simultaneously or form from one another within the 10-day culture period. These findings demonstrate that interleukin-4 is a potent human macrophage fusion factor and that differential regulation of macrophage fusion by interleukin-4 and interferon-gamma may lead to morphological variants of multinucleated giant cells. PMID- 7485415 TI - Dilemmas in counting the homeless: introduction. PMID- 7485416 TI - Housing dynamics of the homeless: implications for a count. AB - Housing dynamics of homeless people limit all efforts to count them. Data on 60 day housing histories from a sample of homeless substance abusers in New Orleans show that homeless people spend relatively few nights in locations that would make them potentially countable, no matter how thorough the enumeration procedure. PMID- 7485417 TI - Looking behind the numbers in counting the homeless: an invited commentary. AB - Many of criticisms of the Census Bureau's S-Night count of the homeless advanced by Wright and Devine are appropriate. However, a more important issue--the lack of agreement in the field on an appropriate conceptual definition of homelessness -undermines their overall critique. PMID- 7485418 TI - Critical factors in counting the homeless: an invited commentary. AB - Several factors affect Wright and Devine's efforts to estimate the size of the homeless population. The most critical ones are the purpose of the estimate, implicit or explicit definitions of homelessness used in the estimate, and the time period covered by the data sets used in the calculations. PMID- 7485419 TI - Definitional quandaries and other hazards in counting the homeless: an invited commentary. AB - Complexities of both definition and dynamics suggest that the unit of analysis in research on homelessness may need rethinking. Wright and Devine's corrective effort can be criticized on the grounds that it inflates the shortcomings of the admittedly flawed 1990 S-Night count, applies relative proportions from an unrepresentative local sample, and smuggles in an overly inclusive notion of homelessness. PMID- 7485420 TI - Lifetime and five-year prevalence of homelessness in the United States: new evidence on an old debate. AB - A 1990 household-based telephone survey of 1,507 people was followed up by surveying a subsample of those interviewed for the initial report. Results showed very high prevalence figures for homelessness, quite close to those found in the earlier survey. Moreover, using explicit and stringent definitions of literal homelessness and doubling up, most periods of homelessness were found to last for more than one month and, particularly with regard to literal homelessness, to involve serious deprivations and violent victimization. PMID- 7485422 TI - Children of addicted mothers: effects of the 'crack epidemic' on the caregiving environment and the development of preschoolers. AB - Comparison of 25 addicted mothers and 25 nonaddicted controls revealed a high incidence of emotional and physical neglect among the addict group, as well as greater tendency toward depression and more chaotic child-rearing environments. A separate study compared the cognitive, language, and emotional development of 20 cocaine-exposed preschool children and 24 nonexposed children. Although few differences in cognitive and language development were found, group differences in emotional and behavioral status, assessed through maternal report, were significant. Directions for future research are suggested. PMID- 7485421 TI - The role of women's therapy groups in the treatment of chemical dependence. AB - The distinctive needs of chemically dependent women are contrasted with those of men and discussed in relationship to traditional mixed-gender therapy groups. A qualitative study of the contrasting experiences of women in mixed-gender and women-only treatment groups confirmed the hypothesis that their treatment needs are better met in the latter, and that these groups are an essential component of programs treating chemically dependent women. PMID- 7485423 TI - Predicting alcohol and drug use in early adulthood: the role of internalizing and externalizing behavior problems in early adolescence. AB - Unique and interactive internalizing and externalizing behavior problems in adolescence were examined in relationship to drug and alcohol use in young adulthood. Results indicated that higher externalizing scores in adolescence predicted more alcohol use in young adulthood for both males and females and also predicted more marijuana and hard drug use for males. Lower internalizing scores were found to predict more marijuana and hard drug use in young adulthood for boys, but not for girls. PMID- 7485424 TI - Early heterosexual debut: situations of sexual possibility during the transition to adolescence. AB - Preadolescent sexuality among urban, minority youth is considered within a normative developmental context. Fourth- and fifth-grade children living in an urban, primarily African-American, low-income neighborhood were interviewed regarding situations of sexual possibility and contigencies of heterosocial and heterosexual behavior. Findings are discussed in light of factors that may influence early heterosexual debut. PMID- 7485425 TI - The role of family and peers in social self-efficacy: links to depression in adolescence. AB - The roles of family, peers, and selected social factors in the origins of depression were assessed in a school-based study of adolescents. Social self efficacy and social support from family and peers were found to be interrelated in their links with depression. Risk factors are delineated and suggestions for preventive intervention are offered. PMID- 7485426 TI - Parents and children sleeping together: cosleeping prevalence and concerns. AB - Review of the cosleeping literature yields a very limited body of prevalence research, only a small portion of which involves psychiatric populations. An archival records review of child and adolescent urban outpatients suggests that prevalence has been underestimated in the literature and that cosleeping is associated with child and parent anxiety and issues of separation and sleep management, rather than with inappropriate sexual contact. PMID- 7485427 TI - Subjective burden among family members of mentally ill adults: relation to stress, coping, and adaptation. AB - Among 225 family members participating in a study of psychoeducational interventions, 39% of the variance in subjective burden was explained by the severity of stressors related to the relative's illness and by mediators related to social support and coping capabilities. Interventions directed toward improving self-efficacy and coping strategies may be important mediators of subjective burden. PMID- 7485428 TI - Rape, torture, and traumatization of Bosnian and Croatian women: psychological sequelae. AB - The first 25 Bosnian women admitted to the Zagreb Obstetrics and Gynaecological Clinic or its associated regional psychiatric centers were assessed using both clinical and post-traumatic stress disorder interviews. Most of the women had been multiply traumatized; all had been repeatedly raped. Psychological status was assessed for those women who were not impregnated, for those impregnated who received abortions, and for those impregnated who carried the fetus to term. PMID- 7485429 TI - Recreational multifamily therapy for troubled children. AB - Recreation-based family groups were formed as modalities in the treatment of school-age children enrolled at a day treatment center and of preschoolers with potential for enrollment. The groups, which had psychoeducational and self-help components, were successful in involving hitherto unwilling parents in their children's treatment, in helping them acquire parenting skills, and in reducing family conflict. PMID- 7485430 TI - Latinos and mutual support group: a case for considering culture. AB - The literature on recognized Latino cultural values and their effect on mental health interventions with Latinos is reviewed, and experiences in implementing school-based mutual support groups for Latino parents are examined. Recommendations are made for incorporating cultural values in support group interventions with Latinos to promote increased participation. Implications for research are discussed. PMID- 7485431 TI - Stress management for psychiatric patients in a state hospital setting. AB - A stress management protocol was adapted for use with schizophrenia patients. Elements included short group sessions, reinforcement of stress reduction skills, modified breathing and relaxation exercises, and capillary blood-flow monitoring for quick feedback. The patients demonstrated ability to participate in the group and to learn certain stress management techniques adapted to their particular needs. PMID- 7485432 TI - Socioecology and the ontogeny of sexual size dimorphism in anthropoid primates. AB - This study examines statistical correlations between socioecological variables (including measures of group composition, intermale competition, and habitat preference) and the ontogeny of body size sexual dimorphism in anthropoid primates. A regression-based multivariate measure of dimorphism in body weight ontogeny is derived from a sample of 37 species. Quantitative estimates of covariation between socioecological variables and this multivariate measure are evaluated. Statistically significant covariation between the ontogeny of dimorphism and socioecological variables, with the possible exception of habitat preference, is observed. Sex differences in ontogeny are lacking in species that exhibit low levels of intermale competition and are classifiable as species with monogamous/polyandrous mating systems. Among dimorphic species, two modes of dimorphic growth are apparent, which seem to be related to different kinds of group compositions. Multimale/multifemale species tend to become dimorphic through bimaturism (sex differences in duration of growth) with minimal sex differences in growth rate. Single-male/multifemale species tend to attain dimorphism through differences in rate of growth, often with limited bimaturism. Measures of intermale competition may also covary with these modes of dimorphic growth, but the relations among these variables are sometimes ambiguous. Correlations between dimorphic growth and behavioral variables may reflect alternative life history strategies in primates. Specifically, the ways in which risks faced by subadult males are distributed and the relations of these risks to growth rates seem to influence the evolution of size ontogenies. The absence of dimorphic ontogeny in some species can be tied to similar distributions of risk in each sex. In taxa that become dimorphic primarily through rate differences in growth, the lifetime distribution of risks for males may change rapidly. In contrast, males may face a pattern of uniformly changing or stable risk in species that become dimorphic through bimaturism. Finally, much variation recorded by this study remains unexplained, providing additional evidence of the need to specially examine female ontogeny before primate body size dimorphism can be satisfactorily explained. PMID- 7485433 TI - Subfossil Indri indri from the Ankarana Massif of northern Madagascar. AB - Subfossil specimens of Indri indri have been recovered recently from the Ankarana Massif cave system in the far north of Madagascar. Taken together with material from the central highland site of Ampasambazimba, the range of this species appears to have once included much of the northern half of the island and to have extended north and west beyond the eastern rainforest (not unlike Hapalemur simus). It is probable that forest corridors connected the subfossil localities to the current range at some time in the past. Climatic desiccation (fluctuating or long-term) and/or human degradation of the environment may have created the disjunct distributions of living and subfossil I. indri. It is also possible that I. indri once included populations or subspecies that were better adapted to dry forest, woodland, or mosaic environments, habitats very different from those occupied by their living conspecifics. Such adaptive diversity would have been similar to that of Propithecus diadema which today has subspecies in the montane forests and one (P.d. perrieri) in the dry forests of the northeast. These discoveries add new information on range extensions to the distributional database for the primates of Madagascar, and illustrate the piecemeal process of their extinctions. PMID- 7485434 TI - Functional anatomy of the trunk musculature in the slow loris (Nycticebus coucang). AB - Lorisid locomotor and postural behaviour exhibits a number of features that distinguish it clearly from other primates. The comparative myological study of the trunk in the slow loris (Nycticebus coucang) and the squirrel monkey (Saimiri sp.) presented here reveals differences that are related to unique aspects of lorisid positional behaviour. While quadrupedal running and leaping requires flexion and extension of the spine, slow climbing quadrupedalism in lorisids depends on spinal lateral flexion and rotation. The contrasting development of the epaxial musculature in the two species dissected reflects these different requirements. Bipedal suspension is a common posture in the lorisids during which rotation and dorsiflexion of the head is made possible by the robustly developed deep, dorsal, cervical musculature. The long lower lever arm in the M. rectus abdominis may play a significant role in the ventroflexion required to regain a quadrupedal stance. PMID- 7485435 TI - Genetic structure of the population of Cabo Verde (west Africa): evidence of substantial European admixture. AB - The population of Cabo Verde was founded in the fifteenth century (1462), on the basis of slaves brought from the West African coast and a few Europeans, mainly from Portugal. The polymorphism of six red cell enzymes (ADA, AK1, ALAD, ESD, GLO1, and PGD) and ten plasma proteins (AHSG, BF, F13A, F13B, GC, HP, ORM, PLG, TBG, and TF) was studied in a sample of 268 individuals from Cabo Verde (West Africa). There is no statistical evidence of genetic heterogeneity between the two groups of islands which constitute the archipelago, Barlavento and Sotavento. The gene frequency distribution observed in Cabo Verde differs, in many markers, from that of West African populations, suggesting an important European influence. The proportion of Caucasian genes in the population of Cabo Verde has been calculated to be M = 0.3634 +/- 0.0510, and the considerable dispersion of the locus-specific admixture estimates seems to indicate random drift has also played a role in the evolution of the allele frequencies in the archipelago. Partition of the variance of the mean estimate in evolutionary and sampling variance shows the evolutionary variance is more than ten times higher than the sampling variance. When dendrograms are constructed on the basis of different genetic distances, the population of Cabo Verde clusters with Afro-Americans, forming a different group from the populations of the African continent. This is interpreted as a consequence of the importance of Caucasian admixture both in Afro-Americans and in the population of Cabo Verde. PMID- 7485436 TI - Postcranial estimates of body weight in Proconsul, with a note on a distal tibia of P. major from Napak, Uganda. AB - A distal tibia of Proconsul major from Napak, Uganda, is described. It is morphologically similar to other Proconsul tibiae, only much larger in size. This specimen and others are used to estimate the body weight of P. major from postcrania for the first time. Body weight is predicted from articular and diaphyseal dimensions using regression equations derived from a modern comparative sample of catarrhine primates. The estimated body weight of P. major based on the Napak tibia is 86.7 kg, whereas two other P. major specimens are smaller, giving a total range of 63.4-86.7 kg and an average of 75.1 kg. The regression equations are also used to predict the body weight of specimens from Rusinga/Mfangano belonging to P. nyanzae and P. heseloni. As the body weight estimates generated here are consistent with previous postcranial-based estimates for Proconsul species, the two sets of estimates are pooled to give means of 10.9 kg for P. heseloni (n = 6) and 35.6 kg for P. nyanzae (n = 12). These findings support the traditional assignment of two species at Rusinga/Mfangano. The postcranial body weight estimates for the three species of Proconsul are compared to body weights estimated from M1 area in order to investigate possible differences in scaling between the teeth and limbs in these species. Despite being based on a smaller sample size, the postcranial estimates clearly differentiate the three taxa, whereas the dental estimates form a more continuous distribution. Molar area overestimates body weight in P. heseloni, indicating that it is megadont compared to a large sample of modern anthropoid primates. In contrast, molar area underestimates body weight in P. Nyanzae and especially P. major, suggesting relative microdonty in these taxa. PMID- 7485437 TI - Status and diet in precontact highland Ecuador. AB - Excavation at the Ecuadorian highland site of La Florida in suburban Quito revealed six deep shaft tombs yielding high-status individuals (n = 9) as well as apparent sacrifices and other low-status individuals (n = 23). Determination of sex and age at death of the recovered skeletal remains resulted in a sample of 32 individuals aged from approximately 7 to 50 years of age. The sample of 18 individuals over the age of 18 years included 14 females and 4 males. Temporally, the remains are assigned to the Chaupicruz Phase (circa 100 to 450 AD) of the Regional Developmental Period. In this study, we analyze stable isotopes of carbon and nitrogen from human bone in order to compare the diets of the high- and low-status individuals. Stable carbon isotope analyses were carried out on preserved protein and biological apatite (bioapatite), and stable nitrogen isotope analyses were carried out on preserved protein. There is a statistically significant difference in delta 13C between the two groups for both protein and mineral sources of carbon with evidence for the greater consumption of maize in the high-status group. There is no significant difference in delta 15N between the two groups, nor is there a significant difference in the spacing between protein and mineral delta 13C values between the two groups. Ethnohistorical evidence for the 16th century AD provides the expectation that the only dietary difference was the higher consumption of animal protein by the elite. There is no evidence for this based on the bone chemistry data from La Florida.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7485438 TI - Effects of body size and social context on the arboreal activities of lowland gorillas in the Central African Republic. AB - The objectives of this 27 month study were to document the positional behaviors used by lowland gorillas (Gorilla gorilla gorilla) in the Central African Republic and to compare the effects of body size dimorphism on the use of arboreal substrates. During this study, despite their great size, all gorillas used trees regularly. Predictions concerning the relationship of body size to arboreal behavior were generally upheld. Small branch and suspensory activities were rare for silverbacks. Females used smaller and multiple substrates and suspensory postures more frequently than males. Although females foraged in the periphery of trees, males stayed close to the cores and rarely used terminal branches. In addition to body size, this study found that party size, social rank, and tree structure all influence an animal's substrate choice and subsequent positional activities. Lone males typically remained in the cores of trees where substrates are large. Group males may have been forced to use all parts of trees because others were present. Lone males used small crown trees which provided easy access to terminal branch foods. Males and females foraging together used larger trees (containing more feeding sites) than single sex groups. Female positional behavior may have been affected by the presence of males. When apart from males, females used the cores of trees and larger substrates more than when foraging with males. As habitat and social context both influence substrate use, they should be considered essential components of body sized based interpretations of the behavior of fossil or extant species. PMID- 7485439 TI - Effect of life history on the squirrel monkey (Platyrrhini, Saimiri) cranium. AB - Among primates, squirrel monkeys uniquely possess an interorbital fenestra, in which the midline bony orbitosphenoid septum is largely absent and the soft tissues of the orbits are separated only by a thin membrane. Neural development may contribute to the approximation of the orbits to the midline in Saimiri, insofar as other platyrrhines with relatively large brains also have relatively narrow interorbital spaces compared to their near relatives. In Saimiri the narrow spacing of the orbits is further exacerbated by intense predation pressure on infants that may select for precocial neonates. The result is a large-headed neonate that is subject to unusual parturition constraints. These parturition constraints apply to the size and dolichocephalic shape of the squirrel monkey head in general, and to the relatively large eyes and approximated orbits in particular. The unique interorbital condition in Saimiri is an example of the effects of life history on skeletal morphology. PMID- 7485440 TI - Changes in transcripts encoding calcium channel subunits of rat myometrium during pregnancy. AB - Extracellular Ca2+ is normally required for myometrial cells to contract. Ca2+ enters muscle cells mainly through voltage-dependent Ca2+ channels (VDCCs) that open in response to action potentials. The synthesis of myometrial VDCCs may change during pregnancy to alter excitation-contraction coupling. We investigated the mRNA levels for the alpha 1- and beta-subunits of the L-type VDCC in rat myometrium to determine whether alterations are associated with term or preterm labor. RNA isolated from myometrial tissues was analyzed by reverse transcription polymerase chain reaction (PCR) using specific primers designed according to the published sequences of the VDCC subunits. From pregnant rat myometrium, two distinct PCR products were obtained for the alpha 1-subunit: one of the expected size at 372 bp and a smaller at 339 bp. Sequence analysis of the larger product revealed a 99.5 or 88% sequence homology between rat myometrium and rat aorta or rabbit heart, respectively, and the smaller product had an identical sequence to a 33-bp deletion. The two alpha 1-products followed the same trend throughout pregnancy. VDCC alpha 1-mRNA levels increased gradually to 6.9-fold just before labor on day 22 but decreased during labor. However, the beta-subunit mRNA level increased sharply on day 22 and then also declined during labor. Progesterone treatment from day 19 to day 22 inhibited term delivery and prevented the significant increase in alpha 1-mRNA levels. In contrast, antiprogesterone (onapristone, ZK-98.299) treatment on day 17 caused a statistically significant increase in the alpha 1- and beta-VDCC subunit mRNA after 8 and 15 h, respectively, then a decrease during preterm labor at 24 h. We conclude that mRNA levels for the VDCC subunits increase before term and preterm labor but decline during periods when VDCCs are likely at their peaks. The increase in levels of mRNA for VDCC likely reflects changes in expression of VDCCs during periods of term and preterm labor that may facilitate uterine contractility required for this process. Progesterone withdrawal or blockade appears to be responsible for regulating levels of mRNA for VDCC in the myometrium in preparation for labor. PMID- 7485441 TI - Protein kinase C in cyclic stretch-induced nerve growth factor production by urinary tract smooth muscle cells. AB - Cyclic stretch of cultured urinary tract smooth muscle cells has been used to mimic some of the events that occur with bladder obstruction. The stretch stimulus induces production of nerve growth factor (NGF), which has been implicated in changes in bladder innervation. Stretch-induced NGF production was blocked by actinomycin. Involvement of protein kinase C (PKC) in the stretch induced NGF production is strongly suggested by the following observations. Phorbol ester activators of PKC mimicked the stretch response as did platelet derived growth factor (PDGF), which acts, in part, through generation of endogenous diacylglycerols. Both stretch- and PDGF-induced NGF production were blocked by prolonged incubation with phorbol ester to downregulate PKC. Western blot analysis confirmed partial downregulation of the Ca(2+)-dependent PKC-alpha and PKC-beta 1 and near complete downregulation of the Ca(2+)-independent PKC isozymes delta, epsilon, and zeta. The involvement of PKC in transducing a physical stimulus (stretch) into a biochemical response (NGF production) has implications for novel types of therapeutic intervention in ailments such as bladder obstruction. PMID- 7485442 TI - Control of beta-myosin heavy chain expression in systemic hypertension and caloric restriction in the rat heart. AB - In the rat left ventricle, both pressure overload induced by abdominal aortic constriction (Abcon) and caloric restriction (CR) induce an increase in the steady-state level of the beta-myosin heavy chain (MHC) protein and mRNA. Both models also induce a concomitant decrease in the alpha-MHC protein and mRNA. The goals of this study were to 1) determine if the changes in MHC expression in the models are due to altered transcription and 2) identify the relative levels of some key factors interacting with the regulatory regions of these genes. Female Sprague-Dawley rats were randomly assigned to the following groups: 1) normal control (NC), 2) Abcon, and 3) CR. After 5 wk of experimental manipulations, myocardial nuclei were isolated. These nuclei were used for 1) nuclear run-on assays or 2) nuclear extract, which was prepared and used for gel mobility shift assays (GMSAs). Nuclear run-on assays demonstrated that the increase in beta-MHC mRNA and protein expression in both Abcon and CR can be at least partially attributed to increased transcription. The concomitant decrease in alpha-MHC content can similarly be attributed to a decrease in transcription of this gene. Furthermore, GMSAs demonstrate that nuclear extract from each group interact differently with certain elements known to be important for expression in vitro. CR nuclear extracts have a 25.6 +/- 7.2% decrease (P < 0.05 vs. NC) in interaction with a thyroid-responsive element, a potential repressor of beta-MHC transcription.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7485443 TI - Immunolocalization of rat cardiac beta-MHC protein expression in systemic hypertension and caloric restriction. AB - Previous studies have shown that both systemic hypertension induced by abdominal aortic constriction (Abcon) and 50% caloric restriction (CR) increase left ventricular (LV) beta-myosin heavy chain (MHC) protein expression in the rat. However, these two physiological states have different effects on hemodynamic load, and information regarding beta-MHC localization across the LV wall in these two models may provide insight into the process of adaptation to chronic stress among myocardial cells. Thus the goal of this study was to determine the pattern of beta-MHC protein expression across the LV wall in Abcon and CR models using a beta-MHC-specific antibody. Adult female Sprague-Dawley rats (approximately 225 250 g) were randomly assigned to one of three groups: 1) normal control (NC), 2) Abcon, and 3) CR. After a treatment period of 5 wk, Abcon LVs hypertrophied 52% relative to NC, accompanying the 42% increase in mean blood pressure. CR rats, however, had a normal LV weight-to-body weight ratio. The relative content of LV beta-MHC protein expression, as assessed by native gel electrophoresis, increased from 3% in NC to 25 and 41% in Abcon and CR rats, respectively. Immunohistochemical analysis of beta-MHC expression demonstrated that the increase in beta-MHC protein in the Abcon group occurred primarily on the endocardial side of the LV. In contrast, the increase in beta-MHC protein in the CR LV occurred equally across the entire LV wall. This suggests that CR has a global effect on MHC isoform expression in LV myocardial cells.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7485444 TI - Specific modulation of cyclic ADP-ribose-induced Ca2+ release by polyamines. AB - Cyclic ADP-ribose (cADPR) is a potent mediator of Ca2+ mobilization from intracellular stores in sea urchin eggs. However, the regulation of the cADPR induced Ca2+ release system is not yet fully elucidated. We now report that spermine and related polyamines, in physiological concentrations, were able to inhibit the Ca2+ release induced by cADPR in sea urchin egg homogenate bioassays, as measured using the Ca2+ indicator fluo 3, but had no effect on the Ca2+ release induced by D-myo-inositol 1,4,5-trisphosphate (IP3) or by nicotinate adenine dinucleotide phosphate (NAADP). Spermine was a more potent inhibitor of the cADPR-induced Ca2+ release than spermidine and putrescine. Spermine inhibited not only the release induced by cADPR but also the Ca2+ release induced by caffeine and ryanodine. Finally, pretreatment of the sea urchin egg homogenates with caffeine or Sr2+ and Ca2+ prevented the inhibitory effect of spermine on cADPR-induced Ca2+ release. We propose that polyamines, which are present in millimolar concentrations in fertilized eggs, are specific inhibitors of the ryanodine channel and perhaps may serve as endogenous regulators of the cADPR induced Ca2+ release system. PMID- 7485445 TI - Polarized rabbit type 1 angiotensin II receptors manifest differential rates of endocytosis and recycling. AB - Receptor-mediated endocytosis and recycling have been described for extrarenal angiotensin II (ANG II) receptors. In proximal tubule (PT) epithelia expressing polarized ANG II receptors, these processes have not been examined as thoroughly. We utilized a PT cell model, LLC-PKCl4 cells stably transfected with rabbit type 1 ANG II receptor (AT1R) cDNA, to investigate these properties. LLC-PK-AT1R cells expressed the rabbit AT1R transcript and displayed losartan-inhibitable specific 125I-labeled ANG II binding at apical (AP) and basolateral (BL) membranes when grown on permeable supports. AP AT1R internalized 125I-ANG II more rapidly than BL AT1R, and phenylarsine oxide treatment inhibited AP AT1R internalization without significantly affecting BL AT1R endocytosis. Pertussis toxin had no effect on AP or BL AT1R endocytosis. In addition, AP AT1R recovered specific 125I ANG II binding after ANG II treatment (a measure of recycling). BL AT1R displayed minimal recovery of 125I-ANG II binding after ANG II pretreatment. These data suggested that AP AT1R enter endocytic/endosomal pathways. Phospholipase A2 (PLA2) activity has been linked to endosomal fusion in other systems, and PT brush-border membrane AT1R also have been associated with PLA2 activity. LLC-PK AT1R cells were therefore treated with quinacrine, a nonspecific PLA2 inhibitor, or Compound I (CI), a selective Ca(2+)-independent PLA2 inhibitor, to determine if PLA2 activity was linked to AT1R recycling. Both quinacrine and CI decreased AP AT1R recycling without affecting BL AT1R recycling. Polarized AT1R in LLC PKCl4 cells thus display differential rates of endocytosis and recycling.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7485446 TI - Altered hepatic localization and expression of occludin after common bile duct ligation. AB - Epithelial tight junctions form a regulated barrier that seals the paracellular space and prevents mixing of luminal contents with the interstitium. This barrier is composed of a group of proteins including the putative "sealing" protein occludin that appears to bind directly to a cytoplasmic junction protein, ZO-1. To study the interaction and regulation of these two components when paracellular integrity is altered, we assessed protein expression and immunofluorescent (IF) localization of ZO-1 and occludin in a rat model of hepatocyte tight junction damage induced by common bile duct ligation (CBDL). Protein levels were detected in liver by immunoblotting and IF localization by 3-dimensional reconstruction of serial 0.5-micron confocal microscopic optical sections. As previously described, ZO-1 protein levels progressively increased to threefold control levels 9 days after CBDL. In contrast, occludin protein levels decreased by 50% within 2 days after CBDL and returned to control values by 9 days. In control IF sections, ZO-1 and occludin colocalized, forming thin continuous staining outlining canaliculi. After CBDL, ZO-1 staining appeared discontinuous, and a punctate pericanalicular accumulation of signal developed around junctional areas. Occludin staining was also discontinuous after CBDL, but, in contrast to ZO-1, was not punctate and remained localized either in a linear fashion along canalicular margins or in a homogeneous fashion in immediately surrounding areas. CBDL results in changes in the expression and localization of the putative tight junction sealing protein occludin in hepatocytes that are distinct from those observed for the peripheral membrane tight junction protein ZO-1.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7485447 TI - Volume-activated chloride channels in HL-60 cells: potent inhibition by an oxonol dye. AB - When swollen in hypotonic media, HL-60 cells exhibit a regulatory volume decrease (RVD) response as a result of net losses of K+ and Cl-. This is primarily caused by a dramatic increase in Cl- permeability, which may reflect the opening of volume-sensitive channels (11). To test this hypothesis, we measured volume activated Cl- currents in HL-60 cells using the patch-clamp technique. The whole cell Cl- conductance (in nS/pF at 100 mV) increased from 0.09 +/- 0.06 to 1.15 +/ 0.19 to 1.64 +/- 0.40 as the tonicity (in mosmol/kgH2O) of the external medium was decreased from 334 to 263 to 164, respectively. Cl- currents showed no significant inactivation during 800-ms pulses. Current-voltage curves exhibited outward rectification and were identical at holding potentials of 0 or -50 mV, suggesting that the gating of the channels is voltage independent. The selectivity sequence, based on permeability ratios (PX/PCl) calculated from the shifts of the reversal potentials, was SCN- > I- approximately NO3- > Br- > Cl- >> gluconate. 4-Acetamido-4'- isothiocyanostilbene-2,2'-disulfonic acid (SITS; 0.5 mM) inhibits HL-60 Cl- channels in a voltage-dependent manner, with approximately 10-fold increased affinity at potentials greater than +40 mV. Voltage-dependent blockade by SITS indicates that the binding site is located near the outside, where it senses 20% of the membrane potential. These Cl- channels were also inhibited in a voltage-independent manner by the oxonol dye bis-(1,3-dibutylbarbituric acid)pentamethine oxonol [diBA-(5)-C4] with a concentration that gives half inhibition (IC50) of 1.8 microM at room temperature. A similar apparent IC50 value (1.2 microM) was observed for net 36Cl efflux into a Cl(-)-free hypotonic medium at 21 degrees C. It seems likely, therefore, that the volume-activated Cl- channels are responsible for the net Cl- efflux during RVD. These Cl- channels have properties similar to the "mini-Cl-" channels described in lymphocytes and neutrophils and are strongly inhibited by low concentrations of diBA-(5)-C4. PMID- 7485448 TI - An oxonol dye is the most potent known inhibitor of band 3-mediated anion exchange. AB - When cells are acutely exposed to the oxonol dye, bis(1,3-dibutylbarbituric acid)pentamethine oxonol (diBA), at 0 degrees C, the concentration that gives half inhibition of Cl- exchange (IC50) is 0.146 +/- 0.013 microM (n = 12) initially, but the inhibition increases with time. These characteristics indicate that a rapid initial binding is followed by a slow conformational change that makes the binding tighter. If diBA is allowed to equilibrate with band 3, the IC50 is only 1.05 +/- 0.13 nM (n = 5), making diBA a more potent inhibitor than 4,4'-diisothiocyanostilbene-2,2'-disulfonic acid (DIDS), for which the IC50 under similar conditions is 31 +/- 6 nM [T. Janas, P. J. Bjerrum, J. Brahm, and J. O. Wieth. Am. J. Physiol. 257 (Cell Physiol. 26): C601-C606, 1989]. Inhibition by diBA is very slowly reversible at 0 degrees C (t1/2 > 50 h), but the effect is more readily reversible at higher temperatures. DiBA competes with 4,4' dinitrostilbene-2,2'-disulfonate (DNDS) for inhibition, suggesting an external site of action. In contrast to DIDS and DNDS, however, increasing Cl- concentrations do not decrease the inhibitory effect of diBA, indicating that the inhibition is not competitive. Thus diBA may be useful for investigating conformational changes during anion exchange and for stopping transport without preventing substrate binding. However, when diBA and other oxonols are used to sense membrane potential, they may have undesirable side effects on anion transport processes. PMID- 7485449 TI - G protein regulation of cardiac muscarinic potassium channel. AB - Several ion channels can be regulated by G proteins in a "membrane-delimited" manner. The cardiac muscarinic K+ (KACh) channel, which is responsible for the acetylcholine (ACh) or adenosine-induced deceleration of heart beat and atrioventricular conduction, is the prototype of this type of receptor-dependent regulation of ion channels. Because similar transduction mechanisms are utilized by various membrane receptors, such as somatostatin, 5-hydroxytryptamine-1, alpha 2-adrenergic, mu-and delta-opioid, D2-dopamine, and gamma-aminobutyric acid B receptors, in neuronal, hormone-secreting, renal, or smooth muscle cells, the G protein (GK)-KACh channel system illustrates the principles underlying one of the most important cell signaling mechanisms (B. Hille. Neuron 9: 187-195, 1992). It seems that both alpha- and beta gamma-subunits of GK may be involved in the regulation of the KACh channel of mammalian atrial muscle. A general consensus of opinion has emerged, after some years of controversy, to support the notion that physiological activation of the channel by GK is the responsibility of the beta gamma-subunits. Recent evidence suggests that the KACh channel interacts with the alpha-subunit in the terminating process of activation. PMID- 7485450 TI - Role of cytochrome P-450 epoxygenase metabolites in EGF signaling in renal proximal tubule. AB - Epidermal growth factor (EGF) is a potent epithelial cell mitogen and induces eicosanoid production in many cell types. The present study examined signaling mechanisms for EGF in the renal proximal tubule, where high concentrations of cytochrome P-450 epoxygenase have been reported. In primary cultures of rabbit proximal tubule cells, EGF (30 nM) increased endogenous epoxyeicosatrienoic acid (EET) levels 5.3 +/- 1.4-fold within 10 min (n = 6). In these cells EGF stimulated [3H]thymidine incorporation was significantly inhibited by the cytochrome P-450 inhibitors ketoconazole or clotrimazole but not by the cyclooxygenase inhibitor indomethacin. In fura 2-loaded proximal tubule cells, EGF caused a concentration-dependent increase in cytosolic Ca2+ concentration ([Ca2+]i), due to Ca2+ influx, which was inhibited by either ketoconazole or SKF 525A but not by indomethacin. Addition of 5,6-EET (0.5 microM) also induced Ca2+ influx in proximal tubule cells, whereas 8,9-11,12-, or 14,15-EET did not. In cells treated with bis(2-amino-5-methylphenoxy)ethane N,N,N',N'-tetraacetic acid tetraacetoxy-methyl ester to chelate [Ca2+]i, EGF-stimulated [3H]thymidine incorporation. These results indicate that EGF increases EET levels in proximal tubule and suggest that 5,6-EET or its metabolites may be a modulator of EGF induced [Ca2+]i increases and involved in mitogenesis. PMID- 7485451 TI - Intracellular calcium signals in response to bradykinin in individual neuroblastoma cells. AB - The Ca indicator fura 2 was used to study the modulation of cytoplasmic Ca by bradykinin (Bk) in single N1E-115 murine neuroblastoma cells. Increases in cytoplasmic Ca in response to Bk were mediated by the B2 receptor subtype. Responses to high concentrations of Bk (1-100 nM) were homogeneous and characterized by a rapidly rising transient that decayed to baseline in the continued presence of agonist, with a half-time of 15 s. Responses to low concentrations of Bk (100-500 pM) were more heterogeneous, with longer latencies and often with oscillations. Pretreatment with thapsigargin for 20 min prevented the Ca response, showing that the Ca change results from intracellular Ca release. Removal of external Ca had little effect on the response to Bk, indicating that the agonist does not activate Ca influx. The extent of Ca release and refilling after Bk was tested with ionomycin. A saturating dose of Bk (20 nM) mobilizes > 90% of stored Ca within 30 s, and this is replaced slowly. Replacement of external Na by N-methyl-D-glucamine to block Na/Ca exchange affected the Ca response, causing decreases in latency and in the period of Ca oscillations and increases in overall duration and peak amplitude of the response. PMID- 7485452 TI - H2O2 activates red blood cell K-Cl cotransport via stimulation of a phosphatase. AB - K-Cl cotransport is involved in volume regulation in a number of cell types. Cell swelling stimulates K-Cl cotransport, probably by inhibition of a volume sensitive kinase. K-Cl cotransport can also be activated by oxidants and thiol reagents. We investigated the effect of H2O2 on K-Cl cotransport of LK sheep red blood cells in an attempt to identify the target of oxidants. H2O2 stimulated K Cl cotransport. The stimulation was virtually abolished by subsequent incubation with calyculin, a protein phosphatase inhibitor. This suggests that H2O2 stimulates a calyculin-sensitive phosphatase and activates K-Cl cotransport by causing a decrease in phosphorylation of the transporter or a regulatory protein. The thiol reagent N-ethylmaleimide, which stimulates K-Cl cotransport, did not stimulate cotransport further in cells with cotransport activated by staurosporine but did stimulate cotransport further in cells with cotransport activated by H2O2. These results suggest that there are at least two distinct phosphorylation sites on the transporter or a regulator. The results also suggest that the phosphatase is associated with the membrane. PMID- 7485453 TI - Chronic enhancement of neuromuscular activity increases acetylcholinesterase gene expression in skeletal muscle. AB - We determined levels of mRNA encoding acetylcholinesterase (AChE) in muscles of rats subjected to chronic enhancement of neuromuscular activation. After 8 wk of voluntary wheel running, extensor digitorum longus (EDL) muscles displayed a 72% increase in total AChE activity as a result of a selective threefold increase in the G4 content. Soleus muscles, on the other hand, exhibited a 30% decrease in A12 while displaying a small (33%) increase in total AChE activity. These enzymatic adaptations were paralleled by increases in the levels of AChE mRNAs in both EDL (32%; P < 0.03) and soleus (42%; P < 0.02) muscles. In addition, compensatory hypertrophy of the plantaris muscle increased total AChE activity by 75%. This change was reflected by an elevation in all AChE molecular forms with A12 (89%) and A8 (179%) showing the most prominent increases. Similar to exercise trained muscles, hypertrophied plantaris muscles displayed an increase in AChE transcripts (25%; P < 0.04). These results indicate that increases in neuromuscular activity modulate expression of the AChE gene in vivo and suggest the involvement of pretranslational regulatory mechanisms in the adaptive response of AChE to enhanced neuromuscular activation. PMID- 7485454 TI - Flow-mediated NO release from endothelial cells is independent of K+ channel activation or intracellular Ca2+. AB - The role of K+ channels and intracellular [Ca2+] in flow-induced nitric oxide (NO) production was investigated in bovine aortic endothelial cells in culture. NO release (measured as nitrite production) and K+ channel activity (measured as 86Rb+ efflux) were measured in cells grown on collagen-coated microcarrier beads and perfused in a column. An eightfold increase in flow produced a rapid (within 1 min), sustained, and reversible sixfold increase in NO release. Efflux of 86Rb+ also increased but rapidly returned to baseline and then transiently decreased when flow was decreased. This was probably due to boundary layer washout rather than to K+ channel activation, because an identical pattern was seen for release of [3H]ouabain. Neither tetraethylammonium nor increasing medium [K+] to block K+ currents prevented flow-induced NO release. Removal of medium Ca2+ or chelation of intracellular Ca2+ also did not block flow-mediated NO release. The results demonstrate that flow rapidly increases NO release from endothelial cells but that this increase in NO release is not dependent on activation of K+ channels or changes in intracellular [Ca2+]. PMID- 7485455 TI - Osmolar changes regulate the paracellular permeability of cultured human cervical epithelium. AB - Extracellular nucleotides induce a biphasic change in the transepithelial electrical conductance (GT) of human cervical cells grown on filters: a rapid increase (phase I) followed by a sustained decrease (phase II). To probe the involvement of the intercellular space, its magnitude was varied by manipulating cell volume through changes in extracellular osmolarity. Under baseline conditions [GT = 115 mS/cm2 (approximately 9 omega.cm2)] and during phase II, hypertonic challenges resulted in an increase in GT (0.98% .mosmol-1.l-1 and 0.73%.mosmol-1.l-1, respectively). However, a hypertonic challenge during phase I decreased GT (-0.16%.mosmol-1.l-1). Hypotonic challenges decreased GT during baseline, phase I, and phase II conditions by -1%.mosmol-1.l-1. Similar trends were observed with regard to pyranine permeability. Reduction of extracellular calcium increased GT, abrogated the phase II effect of extracellular ATP, and reversed the effect of a hypertonic challenge. The additive nature of the permeability changes in response to osmotic challenges and to ATP during phase II suggests that different sites are involved in each response, i.e., the resistance of the intercellular space changes with osmolarity and that of the tight junction during phase II. PMID- 7485456 TI - Effects of okadaic acid and intracellular Cl- on Na(+)-K(+)-Cl- cotransport. AB - The Na(+)-K(+)-Cl- cotransporter of the squid giant axon requires ATP and is inhibited by intracellular Cl- (Cli-) in a concentration-dependent manner ([Cl-]i > or = 200 mM completely inhibits the cotransporter). In the present study we address the question of whether inhibition of cotransport by Cli- is due to a Cl(i-)-induced increase of protein phosphatase activity. Intracellular dialysis was used to apply the phosphatase inhibitor okadaic acid (OKA) under conditions of [Cl-]i at 0, 150, or 300 mM during measurement of cotransporter-mediated unidirectional Cl- influx into axons. At 0 mM [Cl-]i, the application of 250 nM OKA had no effect on the cotransport-mediated Cl- influx when axons were dialyzed with the normal intracellular ATP concentration ([ATP]i = 4 mM). Reduction of [ATP] to 50 microM resulted in a significant decrease of the bumetanide-sensitive CL- influx, which could be partially reversed by OKA treatment. Similarly, in ATP limited axons with [Cl-]i at 150 mM, cotransporter influx was partially stimulated by treatment with OKA. However, axons dialyzed with 300 mM [Cl-]i ([ATP]i = 50 microM) had no measurable cotransport influx, nor was subsequent treatment with OKA able to induce a cotransport-mediated Cl- influx. We conclude that the inhibition of cotransport caused by Cli- is not the result of an increase in the OKA-sensitive protein phosphatase activity. PMID- 7485458 TI - Copper transport and kinetics in cultured C6 rat glioma cells. AB - C6 rat glioma cells accumulate and efflux 67Cu. Both processes showed saturation kinetics with increasing 67Cu concentration. The Michaelis constant (Km) for uptake was 0.63 +/- 0.14 microM; maximum velocity (Vmax) was 3.29 +/- 0.57 pmol Cu.mg protein-1.min-1. The Km for efflux was 0.15 +/- 0.06 microM; Vmax was 1.08 +/- 0.71 pmol Cu.mg protein-1.min-1. p-Chloromercuribenzoate (p-CMB) totally blocked 67Cu efflux but had no effect on Km or Vmax of uptake. Total 67Cu in the cell after 50 min was partitioned equally between particulate and soluble fractions. p-CMB-treated cells accumulated more 67Cu, but < 10% was bound to the particulate (membrane) fraction. Pb also increased 67Cu accumulation without affecting Km and Vmax of 67Cu uptake. These data suggest that carriers for import and export of Cu in C6 cells are distinct or operate in two different cellular environments. Efflux is a sulfhydryl-dependent process subject to inhibition by Pb. The data are consistent with a P-type ATPase in the efflux of Cu from cells and the potential for Pb to inhibit the efflux mechanism. PMID- 7485457 TI - cAMP evokes a rise in intracellular Na+ mediated by Na+ pump inhibition in rat aortic smooth muscle cells. AB - The effect of adenosine 3',5'-cyclic monophosphate (cAMP) on intracellular Na+ concentration ([Na+]i) was studied in primary cultured vascular smooth muscle cells from rat aorta. [Na+]i was measured using digital imaging of cells loaded with the Na(+-)sensitive fluorescent dye sodium-bonding benzofuran isophthalate. The cAMP level was raised by 1) the membrane-permeable cAMP derivative 8 bromoadenosine 3',5'-cyclic monophosphate, 2) the combination of the adenylate cyclase activator forskolin and the phosphodiesterase inhibitor 3-isobutyl-1 methylxanthine, and 3) the beta-adrenoceptor agonist isoproterenol. All three methods caused a dose-dependent continuous rise in [Na+]i during 40-60 min of observations. A rise in [Na+]i may be caused by stimulation of the Na+ influx and/or inhibition of Na+ efflux; therefore, the involvement of both mechanisms was studied. Elevation of the cAMP level had no effect on Na+ influx, measured as the rate of rise of [Na+]i when Na+ efflux was inhibited with 1 mM ouabain. In contrast, elevation of the cAMP level attenuated Na+ efflux, measured as the rate of decline of [Na+]i in Na(+)-loaded cells exposed to Na(+)-free medium. cAMP induced inhibition of Na+ efflux was not observed when the Na+ pump was inhibited; therefore, cAMP inhibits the Na+ pump-mediated component of Na+ efflux. Agents that raise the cAMP level also inhibited, in a dose-dependent fashion, ouabain-sensitive 86Rb uptake in rat aortic rings. The latter observation confirms that the cAMP-induced inhibition of the Na+ pump occurs both in cultured cells and in the native tissue.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7485459 TI - Sulfhydryl oxidation and activation of red cell K(+)-Cl- cotransport in the transgenic SAD mouse. AB - The SAD mouse is characterized by the expression of human SAD hemoglobin (Hb), a super S Hb with a higher tendency to polymerize than HbS due to the presence of two additional mutations, Antilles beta 23Ile and D Punjab beta 121Glu. Monovalent cation transport was studied in erythrocytes from SAD-1 (Hb SAD = 19%) and beta-thal/SAD-1 (Hb SAD = 26%) mice. Erythrocytes containing Hb SAD exhibited dehydration, increased maximal rate of Na(+)-K+ pump, unchanged Rb+ flux via the Gardos channel, and increased K(+)-Cl- cotransport. K(+)-Cl- cotransport was defined as Cl(-)-dependent (substitution with sulfamate or methanesulfonate) okadaic acid-sensitive K+ efflux. Volume regulatory decrease via K(+)-Cl- cotransport was also increased in swollen SAD erythrocytes compared with controls. K(+)-Cl- cotransport was stimulated by staurosporine in all mouse strains, but the extent of stimulation was reduced in beta-thal/SAD-1 mice. Treatment with dithiothreitol reduced K(+)-Cl- cotransport activity in SAD-1 and beta-thal/SAD-1 mice to levels similar to that of control strains, indicating that reversible sulfhydryl oxidation contributes to the activated state of K(+) Cl- cotransport in mouse erythrocytes that express transgenic human Hb SAD. PMID- 7485462 TI - Ion microscopy evidence that La3+ releases Ca2+ from Golgi complex in LLC-PK1 cells. AB - The effect of La3+ on LLC-PK1 cells was investigated by ion microscopy, a mass spectrometry-based technique with a spatial resolution of approximately 0.5 micron. Cells were incubated with LaCl3 for 10 min. (1 mM) or 30 min (0.1 mM), and intracellular calcium distributions were measured with a Cameca IMS-3f ion microscope in cryogenically prepared cells. Compared with control cells, La3+ reduced total calcium in the Golgi complex by > 100 microM in both treatments, whereas other cellular regions, such as the nucleus and cytoplasm, remained largely unchanged. These two treatments were repeated on cells that were preincubated with 1 mM ouabain. The presence of ouabain in the medium increased the loss of calcium from the Golgi by about fourfold compared with the treatments without ouabain. The La3+ effect, therefore, was amplified by ouabain-induced Na+ loading, indicating a possible involvement of a Na+/La3+ exchanger. La3+ was detected within cells and its influx was facilitated by Na+ loading. These results suggest that La3+ may affect cellular calcium homeostasis by actions other than as a simple Ca2+ antagonist. PMID- 7485460 TI - Myocardial adaptation to ischemia by oxidative stress induced by endotoxin. AB - In this study, we examined the effects of oxidative stress adaptation on myocardial ischemic reperfusion injury. Oxidative stress was induced by injecting endotoxin (0.5 mg/kg) into the rat. After 24 h, rats were killed, hearts were isolated, and the effects of ischemia-reperfusion were studied using an isolated working heart preparation. The development of oxidative stress was examined by assessing malonaldehyde production in the heart. The antioxidant defense system was studied by estimating antioxidant enzyme activities and ascorbate- as well as thiol-dependent antioxidant reserve. The results of our study indicated that endotoxin induced oxidative stress within 1 h of treatment; the stress was reduced progressively and steadily up to 24 h. The antioxidant enzymes superoxide dismutase, catalase, glutathione (GSH) peroxidase, and GSH reductase were lowered up to 2 h and then increased. Both thiol- and ascorbate-dependent antioxidant reserve were enhanced, but the enhancement of the former was only transitory. After 24 h, endotoxin provided adequate protection to the heart from the ischemic reperfusion injury, as evidenced by improved left ventricular function and aortic flow. Our results suggest that the induction of oxidative stress by endotoxin induced adaptive modification of the antioxidant defense in the heart, thereby reducing ischemic-reperfusion injury. PMID- 7485461 TI - Nitric oxide enhancement of erythropoietin production in the isolated perfused rat kidney. AB - We have previously reported that nitric oxide (NO) and guanosine 3',5'-cyclic monophosphate (cGMP) may be involved in the regulation of erythropoietin (Epo) production in response to hypoxia both in vivo and in vitro (20). In the present studies, we have used the isolated perfused rat kidney to assess the role of NO in oxygen sensing and Epo production. When arterial PO2 was reduced from 100 mmHg (normoxemic) to 30 mmHg (hypoxemic) in the perfusate of this system, perfusate levels of Epo were significantly increased. This hypoxia-induced increase in Epo production was significantly decreased by the addition of NG-nitro-L-arginine methyl ester (L-NAME; 1 mM) to the perfusates. Hypoxemic perfusion also produced a significant increase, and L-NAME significantly inhibited this increase, in intracellular cGMP levels in the kidney when compared with normoxemic perfused kidneys. Quantitative reverse transcription-polymerase chain reaction also revealed that hypoxemic perfusion produced significant increases in Epo mRNA levels in the kidney, which was blocked by L-NAME. Our findings further support an important role for the NO/cGMP system in hypoxic regulation of Epo production. PMID- 7485464 TI - Na+/H+ exchange modulates acidification of early rat liver endocytic vesicles. AB - Endocytic vesicles are acidified by an electrogenic vacuolar H(+)-ATPase. These studies examined whether rat liver endosomes also exhibit Na+/H+ exchange and whether this transporter alters acidification. Extravesicular Na+ caused saturable proton efflux from acidified endosomes with a Michaelis constant for Na+ of 7.6 mM, whereas an in-to-out Na+ gradient caused endosome acidification without MgATP and accelerated acidification with MgATP. Na(+)-driven proton fluxes were little altered by valinomycin or carbonyl cyanide m chlorophenylhydrazone. Na+/H+ exchange was inhibited by Li+ but was not affected by K+, Cl-, amiloride (1 mM), or 5-(N,N-dimethyl) amiloride (0.1 mM). Na+/H+ exchange was detected in "early" but not in "late" liver endosomes or in lysosomes. These data suggest that early rat liver endosomes exhibit Na+/H+ exchange that, immediately after endosome formation, may accelerate vesicular acidification. Because of its insensitivity to amiloride, this exchanger may be a pharmacologically altered form of Na+/H+ exchanger-1 or a new isoform. PMID- 7485463 TI - Selective stimulation of epithelial cells in colonic crypts: relation to active chloride secretion. AB - Stimulation of Cl secretion by prostaglandin E2 (PGE2) was measured as the short circuit current (Isc) across isolated epithelium of the rabbit distal colon. Cellular morphology of columnar and goblet cells during secretion was monitored using light and electron microscopy. Stimulation by PGE2 altered epithelial cell morphology only by a reduction of vacuolar space in the apical pole of crypt columnar cells, consistent with release of vacuole contents. Imaging of isolated crypts using differential interference microscopy confirmed the release of material from columnar cells during the onset of secretion. Inhibition of Cl secretion with the loop diuretic bumetanide did not block vacuole release. The actin filament-disrupting agent, cytochalasin, reduced the PGE2-stimulated Isc by 40% and blocked emptying of the vacuolar space. These electrical and morphological results indicate that the process of active ion secretion is associated with release of the macromolecular contents from apical vacuoles through a mechanism involving the cytoskeleton. In addition, this relationship supports the concept that vacuolated columnar cells of the crypts of Lieberkuhn are the cell type that secretes Cl in response to PGE2. PMID- 7485465 TI - Expression of sarcoplasmic reticulum Ca2+ transport proteins in cold-acclimating ducklings. AB - The relationship between the cold-induced increase in sarcoplasmic reticulum (SR) Ca2+ transport proteins and the development of muscular nonshivering theermogenesis (NST) was investigated by determining the time course of expression of the sarcoplasmic or endoplasmic reticulum Ca(2+)-ATPase (SERCA), Ca2+ release channel, and calsequestrin in control and cold-acclimating ducklings. 45Ca2+ uptake and [3H]ryanodine binding measurements with skeletal muscle homogenates showed that a cold acclimation period of approximately 4 wk was required to observe a substantial increase in Ca(2+)-ATPase activity and Ca2+ release channel content, which correlates well with NST development Immunoblot analysis of muscle homogenates showed no differences in calsequestrin levels but revealed that the decrease in SERCA2a content was delayed in cold-acclimating birds and that the SERCA1 level was increased after 4 wk of cold acclimation. The persistence of SERCA2a may be related to shivering thermogenesis occurring preferentially in slow-twitch fibers. SERCA1 may account for most of the cold induced increase in 45Ca2+ uptake activity, suggesting the preferential occurrence of a Ca(2+)-dependent NST in fast-twitch fibers. PMID- 7485466 TI - Human vascular smooth muscle cells produce an intracellular form of interleukin-1 receptor antagonist. AB - Interleukin-1 (IL-1) is a proinflammatory monocyte- and macrophage-derived cytokine that has potent vasorelaxant effects on vascular smooth muscle cells (VSMC). VSMC themselves also express both IL-1 alpha- and beta-genes, suggesting that IL-1 may be an autocrine regulator of VSMC function. The present study demonstrates that human saphenous vein VSMC (HSVSMC) produce IL-1 receptor antagonist (IL-1Ra), a specific inhibitor of IL-1 action. IL-1Ra was produced constitutively in most experiments, and its production was upregulated by phorbol 12-myristate 13-acetate and by IL-1 beta. IL-1Ra produced by HSVSMC remained predominately cell associated and was not detectable extracellularly. Furthermore, reverse transcription-polymerase chain reaction analysis and cDNA sequencing indicated that HSVSMC express the alternatively spliced form of IL-1Ra which lacks the signal peptide present in secreted IL-1Ra. HSVSMC also produced IL-1 alpha and the precursor form but not the mature form of IL-1 beta. These results suggest that HSVSMC lack active IL-1 beta-converting enzyme. Like IL-1Ra, IL-1 beta precursor and IL-1 alpha remained cell associated, predominately in the cytosolic fraction. IL-1 beta induced production of both IL-1Ra and IL-1 alpha at each time point and concentration tested. In contrast, platelet-derived growth factor and transforming growth factor-beta augmented production of IL-1Ra, but not that of IL-1 alpha. These results are suggestive of an autocrine role for cell-associated IL-1Ra, as well as for IL-1 alpha and IL-1 beta, in the regulation of VSMC function. PMID- 7485467 TI - Functional characterization of phosphoinositide-specific phospholipase C-beta 1 and -beta 3 in intestinal smooth muscle. AB - Soluble and membrane phosphoinositide-specific phospholipases obtained separately from dispersed circular and longitudinal intestinal muscle cells were characterized for substrate specificity and G protein dependence using selective antibodies to various isoforms of phospholipase C (PLC) and G protein subunits. Western blot analysis disclosed the presence of the main PLC isozymes, PLC-gamma 1, PLC-delta 1, and PLC-beta 1. Soluble PLC from circular and longitudinal muscle was stimulated by guanosine 5'-O-(3-thiophosphate) and inhibited by PLC-beta 1 antibody (80-90%) and PLC-beta 3 antibody (approximately 25%) but not by G protein antibodies. Membrane PLC from circular and longitudinal muscle was stimulated by cholecystokinin octapeptide (CCK-8) and inhibited selectively by PLC-beta 1 antibody (85%), PLC-beta 3 antibody (15%), and G alpha q/11 antibody (90%). CCK-8-induced contraction in permeabilized circular muscle cells was also selectively inhibited by PLC-beta 1 antibody (76%), PLC-beta 3 antibody (24%), and G alpha q/11 antibody (86%). The combined effects of PLC-beta 1 and PLC-beta 3 antibodies on PLC activity and muscle contraction were additive, causing complete inhibition. Soluble and membrane PLC from circular and longitudinal muscle were immunologically similar but functionally different. The enzymes from circular muscle preferentially hydrolyzed endogenous and exogenous phosphatidylinositol 4,5-biphosphate (PIP2), confirming previous findings of preferential hydrolysis of PIP2 in dispersed intestinal circular muscle cells. PMID- 7485468 TI - Calcium requirement for cGMP production during muscarinic activation of N1E-115 neuroblastoma cells. AB - Muscarinic agonists elicit large increases in intracellular Ca2+ and guanosine 3',5'-cyclic monophosphate (cGMP) in N1E-115 neuroblastoma cells. Both signals are blocked in cells loaded with the Ca2+ buffer 1,2-bis(2-aminophenoxy)ethane N,N,N',N'-tetraacetic acid showing that the increase in intracellular Ca2+ concentration ([Ca2+]i) is necessary to stimulate cGMP accumulation. Inhibition of nitric oxide synthase (NOS) blocks the cGMP response without affecting the peak amplitude of the intracellular Ca2+ signal, and it is concluded that Ca(2+) dependent activation of NOS is required for cGMP production. cGMP accumulation is reduced by 60% when cells are bathed in Ca(2+)-free saline, but the peak change in [Ca2+]i is not affected. This suggests that Ca2+ influx is strongly coupled to the activation of cGMP production, even though it makes a smaller contribution to the intracellular Ca2+ signal than does Ca2+ release. Thapsigargin, which releases Ca2+ from intracellular stores, activates Ca2+ influx and increases cGMP. The cGMP increase is transient and follows approximately the same time course as Ca2+ store depletion. Ca2+ influx remains activated after store depletion, however, which indicates that influx alone cannot sustain cGMP production. It is concluded that summation of Ca2+ influx and Ca2+ release is necessary to reach a threshold Ca2+ level needed to stimulate cGMP accumulation. Because of the large contribution from Ca2+ influx, we suggest that NOS or a cofactor necessary for its activation may be located close to Ca2+ channels in the membrane. PMID- 7485470 TI - Human ATP1AL1 gene encodes a ouabain-sensitive H-K-ATPase. AB - The cDNA for ATP1AL1, the fifth member of the human Na-K-adenosinetriphosphatase (ATPase)/H-K-ATPase gene family, was recently cloned (A. V. Grishin, V. E. Sverdlov, M. B. Kostina, and N. N. Modyanov. FEBS Lett. 349: 144-150, 1994). The encoded protein (ATP1AL1) has all the primary structural features common to the catalytic alpha-subunit of ion-transporting P-type ATPases and is similar (63-64% identity) to the Na-K-ATPase alpha-subunit isoforms and the gastric H-K-ATPase alpha-subunit. In this study, ATP1AL1 was expressed in Xenopus laevis oocytes in combination with the beta-subunit of rabbit gastric H-K-ATPase. The functional properties of the stable alpha/beta-complex were studied by 86Rb+ uptake and demonstrated that ATP1AL1 is a novel human K(+)-dependent ATPase [apparent half constant activation/(K1/2) for K+ approximately 375 microM)]. ATP1AL1-mediated inward K+ transport was inhibited by ouabain (inhibition constant approximately 13 microM) and was found to be inhibited by high concentrations of SCH-28080 (approximately 70% at 500 microM). ATP1AL1 expression resulted in the alkalinization of the oocytes' cytoplasm and ouabain-sensitive proton extrusion, as measured with pH-sensitive microelectrodes. These data argue that ATP1AL1 is the catalytic alpha-subunit of a human nongastric P-type ATPase capable of exchanging extracellular potassium for intracellular protons. PMID- 7485471 TI - Responsiveness of mutants of NHE1 isoform of Na+/H+ antiport to osmotic stress. AB - Hypertonic activation of NHE1, the ubiquitous Na+/H+ exchanger, plays a central role in cell volume regulation, yet little is known about the underlying mechanism. We probed the osmotic responsiveness of full-length and truncated constructs of NHE1 transfected into cells lacking endogenous antiport activity. The hypertonic stimulation of NHE1 was preserved after heterologous transfection of the full-length NHE1 or of constructs truncated at positions 698 or 703. In contrast, mutants truncated at position 635 (delta 635) failed to respond to osmotic challenge. Transfectants (delta 635) behaved as if constitutively activated, having a permanently elevated cytosolic pH (pHi) under isotonic, unstimulated conditions. The delta 635 mutant displayed H+ binding with high affinity and low cooperativity. Constructs delta 582 or delta 566 had a reduced H+ sensitivity and were therefore inactive at resting pHi. Such cells were unresponsive to osmotic stress near physiological pHi but could be activated by shrinking after an acid load. Jointly, these results suggest that the H+ affinity and high cooperativity of the antiporter, earlier attributed to a single "modifier site," can be varied independently and are probably controlled by different regions of the molecule. The data indicate that volume or osmolarity sensitive site(s) exist between the NH2-terminus and residue 566. This putative volume-sensitive site is therefore different from the site(s) postulated to mediate the stimulatory effects of calcium and growth factors. PMID- 7485469 TI - Inhibition of MAP kinase by prostaglandin E2 and forskolin in rat renal mesangial cells. AB - Activation of the mitogen-activated protein (MAP) kinase pathway is believed to play a critical role in normal and pathophysiological proliferation of mesangial cells. Recent studies have shown that MAP kinase activation by growth factors in other cell types involves activation of the low-molecular-weight G protein Ras and the protooncogene serine kinase c-Raf-1. In this study, the role of this pathway in rat renal mesangial cells was assessed. Platelet-derived growth factor (PDGF), epidermal growth factor (EGF), as well as phorbol esters (PMA) rapidly activated MAP kinase three- to fourfold in these cells. PDGF and EGF, but not PMA, were able to activate c-Raf-1 and Ras activity. Stimulation of mesangial cells with the inflammatory mediator prostaglandin E2 (PGE2) or elevation of intracellular adenosine 3',5'-cyclic monophosphate (cAMP) by treatment with forskolin markedly blunted activation of MAP kinase induced by PDGF and EGF, but not by PMA. Consistent with this observation, PGE2 abolished growth factor induced activation of c-Raf-1. However, Ras activation induced by growth factors was not affected by PGE2 and forskolin. These results suggest that MAP kinase activation can occur by at least two separate pathways in mesangial cells. Tyrosine kinase receptors activate MAP kinase through activation of Ras and Raf. This pathway can be blocked by PGE2 and elevation of cAMP, presumably by interfering with the ability of Ras to activate Raf. In addition, activation of protein kinase C by phorbol esters can activate MAP kinase in a Ras/Raf independent manner.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7485472 TI - Gene-targeting and transgenic approaches to IGF and IGF binding protein function. AB - The ability to manipulate genetic information in the germ line of mice has provided powerful approaches to study gene function in vivo. These approaches have included the establishment of mouse lines in which a specified gene or genes are overexpressed, ectopically expressed, or deleted. Transgenic and gene targeted mouse lines have been used extensively to study the function of the insulin-like growth factors (IGF), IGF-I and IGF-II, and their receptors and binding proteins. In the IGF system, these technologies have elucidated the roles of the IGFs in fetal and somatic growth and have demonstrated a critical role for this system in transformation and tumorigenesis. Analysis of combinatorial crosses of gene-targeted mouse lines also has suggested the existence of an as yet unidentified IGF receptor that regulates fetal growth. Similar approaches using transgenic and gene-targeted mouse models have been initiated to study the in vivo functions of the IGF binding proteins. These mouse models provide important tools to test specific functional questions in vivo as well as to study the long-term physiological consequences of chronic gene alterations. PMID- 7485473 TI - Evidence that increased glucose cycling in islets of diabetic ob/ob mice is a primary feature of the disease. AB - Glucose cycling (GC) is increased in pancreatic islets from hyperglycemic 6-mo old ob/ob mice. We determined whether normalization of glycemia alters islet GC and insulin release in response to glucose as well as oxidation and utilization of the glucose. Mice were treated with phlorizin in dimethyl sulfoxide (DMSO) for 10 days, which resulted in normalization of blood glucose concentrations. Controls received DMSO. The mice were fasted overnight and killed, and their islets were isolated for measurements of insulin release at 5.5 and 16.7 mM glucose and at 16.7 mM glucose plus 10 mM arginine. GC was measured by the incorporation of 3H from 3H2O into carbon 2 of glucose, glucose oxidation by the yield of 14CO2 from [U-14C]glucose, and glucose utilization by the yield of 3H2O from [5-3H]glucose. Phlorizin treatment did not alter the response of insulin to glucose and to glucose plus arginine. GC was 30% in control and phlorizin-treated animals. Glucose oxidation and utilization were also the same in both groups. In fed 10- to 12-mo-old mice exhibiting a broad range of blood glucose levels, there was no correlation between GC and either insulin release or glucose concentrations. Thus the islets of ob/ob mice exhibit an increased rate of GC regardless of glycemia. This indicates that the increased rate of GC is an important characteristic of the diabetic syndrome in these animals and not simply secondary to hyperglycemia. PMID- 7485474 TI - Tissue IGF-I protein and mRNA responses to a single injection of somatotropin. AB - Swine were divided into four groups of 11 animals at 40 kg body wt. Swine within a group were given a single porcine somatotropin (pST) injection (200 micrograms/kg) or buffer at 0800. Blood, liver (L), latissimus dorsi (LD), semitendinosus (STS), vastus lateralis (VL), dorsal subcutaneous (SQ), and perirenal (PR) adipose tissues were sampled at 0, 1, 2, 4, 8, 12, 16, and 24 h postinjection. Blood urea nitrogen was depressed by 16 h. Insulin was elevated by approximately 350% at 8 h. Lipogenic enzyme activities in adipose tissues were not affected by pST treatment. Insulin-like growth factor I (IGF-I) mRNA levels increased rapidly in SQ, PR, and L to a single pST administration, whereas they increased only slightly in VL. IGF-I mRNA concentrations in LD and STS were unaffected by pST treatment. IGF-I protein content of tissues changed little during the first 24 h postinjection. These data suggest that individual tissues differ in timing and degree of response to pST. Conflicting results reported after pST treatment could, in part, be due to tissue selection for sampling or sample timing. PMID- 7485475 TI - Protein and glucose metabolism during isolated closed-head injury. AB - Patients with isolated closed-head injuries are characterized by excessive nitrogen loss, hyperglycemia, and increased caloric demand. However, the relative contributions of specific metabolic events such as protein breakdown and synthesis or glucose production and utilization to the physiological changes prevalent during isolated closed-head injury remains unestablished. By use of isotopic dilutions of L-[1-13C]leucine and D-[3-3H]glucose, components of protein and glucose metabolism were examined in patients with isolated closed-head injury (n = 7). Normal overnight-fasted volunteers (n = 8) were also studied as a reference point for comparison. Despite prevailing hyperinsulinemia (29 +/- 5 microU/ml), head-injured patients had elevated plasma leucine concentrations (183 +/- 22 vs. 144 +/- 8 mumol/l), whole body proteolysis (331 +/- 44 vs. 150 +/- 7 mg.kg-1.h-1), protein synthesis (248 +/- 38 vs. 126 +/- 11 mg.kg-1.h-1), and amino acid oxidation (84 +/- 11 vs. 23 +/- 3 mg.kg-1.h-1). Therefore nitrogen loss normally associated with isolated closed-head injury is primarily due to an increase in the rate of whole body proteolysis, with a greater proportion of the resultant amino acids being oxidized for energy. Furthermore, head-injured patients were hyperglycemic (6.7 +/- 0.3 mumol/l) with increased rates of glucose turnover (an estimate of production and utilization) compared with the controls (4.0 +/- 0.7 vs. 2.5 +/- 0.2 mg.kg-1.min-1). Hence, these data suggest that head injury, even in the absence of peripheral trauma, induces a physiological state of accelerated metabolism associated with resistance to insulin action. PMID- 7485476 TI - Dexamethasone enhances accumulation of cholesteryl esters by human macrophages. AB - The effects of dexamethasone on lipid accumulation by human monocyte-derived macrophages were investigated. Preincubation of macrophages with dexamethasone for a period of 16-20 h resulted in a reproducible increase (3.5-fold) in the incorporation of oleate into cholesteryl esters. The effect was specific because no alterations were observed in oleate incorporation into triglycerides or phospholipids. Measurement of cellular cholesteryl esters indicated a fourfold increase after preincubation with dexamethasone. This increase was mediated by opposite effects on synthesis and breakdown of these lipids. Dexamethasone produced a 60% increase in activity of the enzyme acyl-CoA: cholesterol O acyltransferase (ACAT), active in synthesis of cholesteryl esters, and a 40% decrease in that of neutral cholesteryl esterase, active in cholesteryl ester breakdown. The increased ACAT activity appeared to reflect increased mRNA for the enzyme. The effects of dexamethasone on cholesteryl ester accumulation by macrophages reached statistical significance at a concentration of 100 nM. They were dose dependent, and saturation was observed at around 1 microM. The effects were significant at low concentrations of cholesterol in the medium. At high medium cholesterol, there was a large cholesterol-induced increase in ACAT activity that obscured most of the effect of dexamethasone. In general, the data suggest that high glucocorticoid levels enhance lipid accumulation by macrophages and thus would have an atherogenic action that is independent of serum cholesterol. PMID- 7485477 TI - A new correction factor for use in tracer estimations of plasma fatty acid oxidation. AB - The purpose of this study was to acquire a new correction factor for use in tracer estimations of plasma fatty acid oxidation that would fully account for label fixation during the infusion of fatty acid tracers. Thus volunteers were infused with 13C-labeled fatty acids and [1-14C]acetate in the basal state, during hyperinsulinemia-hyperglycemia (clamp), and during 1 h of cycling exercise. The fractional recovery of acetate label (i.e., the acetate correction factor) was 0.56 +/- 0.02, 0.50 +/- 0.03, and 0.80 +/- 0.03 in the basal state and during the clamp and exercise, respectively. Isotopically determined plasma fatty acid oxidation rates (mumol.kg-1.min-1) were 1.7 +/- 0.2, 0.8 +/- 0.2, and 6.4 +/- 0.5 (no correction); 2.1 +/- 0.2, 1.0 +/- 0.2, and 6.7 +/- 0.5 (bicarbonate correction); and 3.1 +/- 0.2, 1.5 +/- 0.2, and 8.2 +/- 0.4 (acetate correction). We conclude that use of the acetate correction factor in place of the bicarbonate correction factor should improve the accuracy of isotopic measurements of plasma fatty acid oxidation, because it accounts for label fixation that might occur at any step between the entrance of labeled acetyl-CoA into the tricarboxylic acid cycle until the recovery of label in breath CO2. PMID- 7485478 TI - Determinants of cardiac fibrosis in experimental hypermineralocorticoid states. AB - Uninephrectomized rats maintained on 1.0% NaCl to drink and infused with aldosterone (0.75 microgram/h) for 8 wk have previously been shown to develop hypertension, cardiac hypertrophy, and cardiac fibrosis. In the present study we have shown that K+ supplementation (1.0% NaCl plus 0.4% KCl drinking solution) alters neither the interstitial nor the perivascular fibrotic response to mineralocorticoid treatment. Second, rats receiving 0.75 microgram/h 9 alpha fluorocortisol, a mineralocorticoid and glucocorticoid agonist, respond with hypertension and cardiac fibrosis without cardiac hypertrophy. Finally, intracerebroventricular infusion of the mineralocorticoid receptor antagonist RU 28318 blocks blood pressure elevation, but not cardiac hypertrophy or fibrosis, when aldosterone is coinfused peripherally. We conclude that the myocardial fibrosis observed in response to chronic mineralocorticoid elevation and salt loading is a humorally mediated event independent of hypokalemia, hypertension, and cardiac hypertrophy. It remains to be determined whether the fibrosis observed in the presence of excess salt represents a direct (e.g., cardiac) effect of mineralocorticoid hormones or one mediated via a primary action on classical epithelial aldosterone target tissues (e.g., kidney). PMID- 7485479 TI - Response of glutamine metabolism to exogenous glutamine in humans. AB - To determine whether exogenous glutamine affects whole body glutamine metabolism, preliminary experiments were performed to verify that L-[1-13C]-, L-[U-14C]-, and L-[3,4-3H]glutamine given simultaneously by vein provided similar estimates of glutamine appearance rates [Ra; 355 +/- 24, 373 +/- 19, and 393 +/- 24 (SE) mumol.kg-1.h-1, respectively, P = NS] in six healthy men; glutamine oxidation accounted for 32 +/- 3 and 51 +/- 5% (P < 0.01) of glutamine Ra when it was measured using L-[U-14C]- and L-[1-13C]glutamine, respectively. Five subjects received two 5-h intravenous infusions of L-[3,4-3H]glutamine and a simultaneous nasogastric infusion of L-[1-13C]glutamine on 2 separate days in the postabsorptive state, along with saline on 1 day and natural L-glutamine (856 +/- 45 mumol.kg-1.h-1) on another day in a randomized order. Splanchnic glutamine extraction (determined from [13C]glutamine appearance into systemic blood) reached 74 +/- 4 and 53 +/- 5% during the enteral infusion of tracer alone and in combination with a large load of glutamine, respectively. Glutamine infusion was associated with increased plasma glutamine concentration (from 630 +/- 50 to 1,297 +/- 75 microM), Ra (from 258 +/- 20 to 589 +/- 45 mumol.kg-1.h-1), and oxidation (from 179 +/- 20 to 477 +/- 47 mumol.kg-1.h-1, all P < 0.01), no change in glutamine release from proteolysis, and a decline in glutamine de novo synthesis (from 156 +/- 15 to 93 +/- 13 mumol.kg-1.h-1).(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7485481 TI - Lactate transport studied in sarcolemmal giant vesicles from rat skeletal muscles: effect of denervation. AB - The effect of denervation on lactate transport capacity was studied in giant sarcolemmal vesicles obtained from rat muscle. The rate of lactate transport was determined in soleus and red (RG) and white gastrocnemius (WG) after 1, 3, and 21 days of denervation and in the corresponding contralateral muscles. In addition, muscle lactate dehydrogenase (LDH) and succinate dehydrogenase (SDH) activities were determined. After 1, 3, and 21 days of denervation the rate of lactate transport was lower (P < 0.05) in WG (9, 11, and 36%), RG (15, 21, and 41%), and soleus (12, 24, and 50%) compared with the contralateral muscles. After 21 days of denervation LDH activity was 26, 25, and 34% and SDH activity 33, 25, and 27% lower (P < 0.05) in WG, RG, and soleus, respectively, compared with the contralateral muscles. In the control muscles the lactate transport capacity was 20 and 32% lower (P < 0.05) in WG than in RG and soleus, respectively. The present findings provide support that the sarcolemmal lactate carrier is a plastic system; the transport capacity in soleus, RG, and WG already declines after 1 day of denervation and is further reduced after 21 days of denervation. In addition, the data suggest that the lactate transport capacity in fast-twitch glycolytic fibers < fast-twitch oxidative-glycolytic fibers < slow-twitch oxidative fibers. PMID- 7485482 TI - In vivo evidence of a positive role for somatostatin to optimize pulsatile growth hormone secretion. AB - Despite convincing evidence that somatostatin (SRIF) and growth hormone (GH) releasing factor (GRF) individually play crucial roles in GH regulation, the nature of the interplay between these two hypothalamic hormones is far from clear. In the present study, we used the long-acting SRIF analogue, octreotide, as a probe in both the normal and mutant dwarf (dw) rat 1) to further elucidate the temporal nature of the SRIF-GRF interaction in vivo and 2) to define possible mechanisms of action of SRIF in generating pulsatile GH secretion. Normal free moving adult male rats pretreated with octreotide (25 and 50 micrograms iv) and subsequently challenged with GRF (1 micrograms iv) exhibited a markedly blunted GH response to exogenous GRF 1 h after treatment. In contrast, preexposure to octreotide for 3 h produced a two- to threefold augmentation in GH responsiveness to GRF. Compared with normal saline-pretreated controls, 3-h pretreatment with octreotide produced a 14- to 16-fold augmentation in the postinhibitory rebound release of GH after the coadministration of native SRIF-14 and GRF (P < 0.001). In dw rats, which show a selective reduction in GH synthesis and storage, 3-h preexposure to octreotide failed to significantly alter GRF-induced GH release. These results demonstrate that, in the normal male rat, a 3-h period of exposure to the SRIF analogue octreotide is sufficient to enhance GH responsiveness to GRF. Our findings suggest that this effect is due to a SRIF-mediated buildup of pituitary GH stores in a readily releasable poo.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7485480 TI - Effects of caffeine on energy metabolism, heart rate, and methylxanthine metabolism in lean and obese women. AB - The magnitude of coffee-induced thermogenesis and the influence of coffee ingestion on substrate oxidation were investigated in 10 lean and 10 obese women, over two 24-h periods in a respiratory chamber. On one occasion the subjects consumed caffeinated coffee and on the other occasion, decaffeinated coffee. The magnitude of thermogenesis was smaller in obese (4.9 +/- 2.0%) than in lean subjects (7.6 +/- 1.3%). The thermogeneic response to caffeine was prolonged during the night in lean women only. The coffee-induced stimulation of energy expenditure was mediated by a concomitant increase in lipid and carbohydrate oxidation. During the next day, in postabsorptive basal conditions, the thermogenic effect of coffee had vanished, but a significant increase in lipid oxidation was observed in both groups. The magnitude of this effect was, however, blunted in obese women (lipid oxidation increased by 29 and 10% in lean and obese women, respectively). Caffeine increased urinary epinephrine excretion. Whereas urinary caffeine excretion was similar in both groups, obese women excreted more theobromine, theophylline, and paraxanthine than lean women. Despite the high levels of urinary methylxanthine excretion, thermogenesis and lipid oxidation were less stimulated in obese than in lean subjects. PMID- 7485483 TI - Blunting of the immediate-early gene and mitogenic response in hepatectomized type 1 diabetic animals. AB - Studies suggest that liver regeneration is delayed in insulin-deficient animals, but defining a role of insulin as a growth factor in hepatic regeneration has remained elusive. By examining gene expression of hepatectomized liver in type 1 diabetic BB rats, we have identified dramatic changes in the expression of primary or immediate-early growth response genes compared with normal animals. These include altered expression of insulin-regulated genes such as glucose-6 phosphatase (G-6-Pase), phosphoenolpyruvate carboxykinase (PEPCK), and beta actin, and genes such as CL-6 and map kinase phosphatase-1 (MKP-1) that were previously unlinked to insulin action in animals. Abnormal elevation of mRNAs encoding G-6-Pase, MKP-1, and PEPCK in the time 0 diabetic liver results in decreased induction after partial hepatectomy. Other genes, such as CL-6 and beta actin, are induced at a lower level in the hepatectomized diabetic animals. The net effect is a blunting of the immediate-early gene response after partial hepatectomy in diabetic animals. As determined by DNA synthesis assays, the regenerative capacity of insulin-deficient BB diabetic livers is reduced, and this defect is corrected at least in part by insulin therapy. These findings suggest that because of insulin deficiency, common intracellular signaling pathways that are required for both metabolism and mitogenesis are aberrant in the type 1 diabetic liver and, as a result, the regenerative response is deficient. PMID- 7485484 TI - Regulation of hexokinase II and glycogen synthase mRNA, protein, and activity in human muscle. AB - Insulin regulates the activity of key enzymes of glucose metabolism in skeletal muscle by altering transcription or translation or by producing activity-altering modifications of preexisting enzyme molecules. Because of the small size of percutaneous muscle biopsies, these phenomena have been difficult to study in humans. This study was performed to determine how physiological hyperinsulinemia regulates the activities of hexokinase (HK), glycogen synthase (GS), and GLUT-4 in human skeletal muscle in vivo. We determined mRNA abundance, protein content, and activities for these proteins in muscle biopsies before and after a hyperinsulinemic clamp in normal subjects. HK I, HK II, GS, and GLUT-4 were expressed in muscle. HK II accounted for 80% of total HK activity and was increased by insulin from a basal value of 2.11 +/- 0.26 to 3.35 +/- 0.47 pmol.min-1.mg protein-1 (P < 0.05); HK I activity was unaffected. Insulin increased GS activity from 3.85 +/- 0.82 to 6.06 +/- 0.49 nmol.min-1.mg-1 (P < 0.01). HK II mRNA was increased 3.3 +/- 1.3-fold (P < 0.05) by insulin infusion. HK I, GS, and GLUT-4 mRNA and protein were unaffected. Because insulin infusion increased HK II but not GS mRNA, we conclude that HK II and GS may be regulated by insulin by different mechanisms in human skeletal muscle. PMID- 7485485 TI - Insulin resistance after hypertension induced by the nitric oxide synthesis inhibitor L-NMMA in rats. AB - To explore the relationship between insulin resistance and hypertension, we examined whether acute induction of hypertension can engender insulin resistance. For this purpose we measured rates of insulin-mediated glucose uptake in awake unstressed rats with the euglycemic hyperinsulinemic (12 microns.kg-1.min-1) clamp technique during infusions of saline alone or after induction of hypertension by bolus administration of NG-monomethyl-L-arginine (L-NMMA, 30 and 15 mg/kg), a competitive inhibitor of nitric oxide synthase. Arterial pressure was approximately 20% greater with L-NMMA bolus than with saline alone. Isotopically determined steady-state rates of glucose uptake were 36 +/- 1 mg.kg 1.min-1 during saline alone and 26 +/- 2 and 19 +/- 1 mg.kg-1.min-1 with low- and high-dose L-NMMA (P < 0.001 vs. saline), respectively. To rule out that insulin resistance induced by L-NMMA was adrenergically mediated, clamp studies were repeated with alpha- and beta-blockade. Rates of glucose uptake remained approximately 20% below those observed with saline alone (P < 0.001). A significant inverse correlation was observed between the height of the blood pressure and the rate of glucose uptake (r = 0.32, P = 0.04). In conclusion, acute induction of hypertension with L-NMMA can cause marked insulin resistance. We postulate that reduced skeletal muscle perfusion and/or sympathetic nervous system activation may contribute to insulin resistance induced by L-NMMA. PMID- 7485486 TI - Identification of target cells for growth hormone's action in the arcuate nucleus. AB - Growth hormone (GH) participates in the regulation of its own secretion by acting through a short-loop feedback mechanism to regulate the synthesis and secretion of somatostatin (SS) and growth hormone-releasing hormone (GHRH). The mechanism of GH's action in certain peripheral targets involves the induction of c-fos. Similarly, we hypothesized that GH induces the expression of c-fos mRNA in SS and GHRH neurons in the hypothalamus. Using in situ hybridization, we observed a significant induction of c-fos mRNA in the arcuate nucleus of human GH-treated compared with control animals. Contrary to our hypothesis, only 11% of GHRH mRNA containing and 5% of SS mRNA-containing neurons colabeled for c-fos mRNA. These findings indicate that GH feedback on the hypothalamus includes the induction of c-fos mRNA primarily in neurons other than GHRH and SS in the arcuate nucleus and suggest that these unidentified neurons located in the arcuate nucleus are directly involved in transducing the effects of GH in the brain. PMID- 7485487 TI - Insulin regulates circulating insulin-like growth factors and some of their binding proteins in lactating cows. AB - Four lactating Holstein cows were subjected to a hyperinsulinemic-euglycemic clamp to evaluate the effects of insulin on circulating concentrations of insulin like growth factors (IGF) and their binding proteins (IGFBP). Baseline blood samples were taken every 4 h for 2 days. For the 4-day clamp, insulin was infused (1 microgram.kg body wt-1.h-1) into the jugular vein and exogenous glucose was infused to maintain euglycemia. Circulating insulin was increased approximately fivefold, while glucose was maintained within 10% of baseline concentrations by infusion of 0.15 g.kg body wt-1.h-1 glucose. Hyperinsulinemia-euglycemia approximately doubled IGF-I (145 vs. 286 ng/ml, SE = 20) while decreasing circulating IGF-II (285 vs. 180 ng/ml, SE = 32). Densitometry of Western blots demonstrated no change in IGFBP-3 or a 30,000 relative molecular weight (M(r)) band during the clamp. However, IGFBP-2 decreased 73% and a 26,000 M(r) band decreased 58% by the end of the clamp. Therefore, insulin, directly or via secondary changes, increased circulating concentrations of IGF-I while decreasing concentrations of IGF-II, IGFBP-2, and a 26,000 M(r) IGFBP in lactating cows. PMID- 7485488 TI - Persistence of metabolic consequences in the progeny of rats fed a HC formula in their early postnatal life. AB - First-generation (1-) male and female rat pups were either reared artificially on a high-carbohydrate (HC) or a high-fat (HF) formula or nursed by mother (MF) from day 4 and weaned onto a stock diet on day 24. 1-HC rats compared with sex-matched control rats (1-HF and 1-MF) were hyperinsulinemic and mildly obese by day 60. We investigated the effect of maternal hyperinsulinemia on the second generation (2 ) by intragroup breeding. The 2-HC male and female rats were hyperinsulinemic on day 45, had significantly increased growth rate from approximately day 60 onward, and became obese as evidenced by increased adipose tissue mass due to hypertrophy on day 100. The lipogenic capacity of liver and adipose tissues was significantly higher in the 2-HC than in control rats. Thus the metabolic changes that occurred in the first-generation rats fed a HC formula during early postnatal life not only persisted into their adult life but were also passed on to the next generation. PMID- 7485489 TI - Neonatal rat dietary carbohydrate affects pancreatic islet insulin secretion in adults and progeny. AB - Neonatal rat pups were artificially reared on isocaloric diets high in carbohydrate (HC) or high in fat (HF) or were naturally reared on mother's milk (MF). The HC adult rats were hyperinsulinemic, normoglycemic, and obese. This study investigates pancreatic islet insulin release (IR) of the adult first generation (1-) diet-regulated animals and their second-generation (2-) progeny. Male rat 1-HC islets had higher basal IR than either 1-MF or 1-HF control groups. In addition, glucose (17 mM) failed to increase IR above basal values in 1-HC islets, whereas it stimulated IR in 1-MF and 1-HF islets. Similar secretory responses were evoked by 2-ketoisocaproic acid (2-KIC). Female rat 1-MF and 1-HF islets also had higher glucose-stimulated IR compared with 1-HC islets. Male rat 2-HC islets had higher basal IR and reduced sensitivity to glucose and 2-KIC compared with 2-MF islets, which coincided with hyperinsulinemia. Glyceraldehyde 3-phosphate dehydrogenase activity in 1-HC and 2-HC islets was higher than in MF islets. These data suggest that basal IR is higher in islets isolated from animals reared as neonates on a diet high in carbohydrate. Alterations in beta cell metabolism and secretion probably contribute to the hyperinsulinemia, reduced glucose sensitivity, and glucose intolerance characteristic of this rat model. PMID- 7485490 TI - Magnesium supplementation reduces development of diabetes in a rat model of spontaneous NIDDM. AB - We examined the effects of a magnesium-supplemented (Mg-S) diet in the male obese Zucker diabetic fatty rat, a model of non-insulin-dependent diabetes mellitus (NIDDM). Obese rats were maintained on either a control (0.20% Mg) or magnesium supplemented (Mg-S; 1% Mg) diet for 6 wk beginning at 6 wk of age. The rats maintained on the Mg-S diet had markedly lower fasting and fed-state blood glucose concentrations and an improved glucose disposal. By 12 wk of age, all of the eight animals on the control diet became diabetic, whereas diabetes developed in only one of eight animals on the Mg-S diet. Insulin and C-peptide concentrations, in addition to pancreatic GLUT-2 and insulin mRNA expression, were higher in the male obese Mg-S rats than in their control-fed counterparts. A subgroup of rats on the control diet with established diabetes was switched to a Mg-S diet for an additional 4 wk. The Mg-S diet did not reverse diabetes once already established. These data indicate that an increased dietary Mg intake in male obese rats prevents deterioration of glucose tolerance, thus delaying the development of spontaneous NIDDM. PMID- 7485491 TI - Effects of a continuous lipid infusion on glucose metabolism in critically ill patients. AB - The effects of lipid administration on carbohydrate oxidation rate remain controversial, particularly in critically ill patients. The aim of this study was to determine the effects of these patients of a continuous lipid infusion on glucose metabolism using indirect calorimetry and stable isotopes. We studied seven patients, mechanically ventilated, during two consecutive 24-h periods. Throughout the first period they received a continuous infusion of glucose (2 mg.kg-1.min-1) and amino acids. During the second period, in addition to the glucose, they received a continuous infusion of 1 mg.kg-1.min-1 of long-chain triglycerides emulsion. Substrate oxidation rates were calculated from pulmonary gas exchange and nitrogen excretion measurements. Glucose kinetic parameters were measured using primed constant infusions of [6,6-2H2]glucose and [1-13C]glucose. The lipid infusion did not modify the glucose metabolism parameters; 45% of the lipid supply was stored. PMID- 7485493 TI - Hepatic glycogen accurately reflected by acetaminophen glucuronide in dogs refed after fasting. AB - To validate a method to "biochemically biopsy" the immediate precursor of intrahepatic glycogen [uridyl diphosphate (UDP)-glucose] using acetaminophen and to assess how fasting affects the direct and indirect pathways of glycogen synthesis, dogs were fasted overnight (group 1, n = 5) or for 2.5 days (group 2, n = 5) and then given a 4-h duodenal infusion of unlabeled glucose, [3 3H]glucose, and [U-14C]lactate to label hepatic glycogen via the direct and indirect pathways, respectively, and [1-13C]galactose to measure intrahepatic UDP glucose flux. After 3 h for equilibration, acetaminophen was given and urine was collected for acetaminophen glucuronide. Multiple liver biopsies were obtained. The mean 3H/14C ratios of glucose derived from glycogen (10.4 +/- 4.1 and 1.1 +/- 0.3 for groups 1 and 2, respectively) and glucose derived from acetaminophen glucuronide (11.5 +/- 4.0 and 1.0 +/- 0.1 for groups 1 and 2, respectively) were similar. Fasting significantly increased UDP-glucose flux, the rate of glycogen synthesis, and the contribution of the indirect pathway. We conclude that, in dogs, 1) no functional hepatic zonation exists with regard to acetaminophen glucuronidation and liver glycogen synthesis and 2) with appropriate choice of isotopic tracers and study design, UDP-glucose flux can accurately reflect rates of hepatic glycogen synthesis. PMID- 7485492 TI - Compensatory alterations for insulin signal transduction and glucose transport in insulin-resistant diabetes. AB - Insulin binding activates the receptor tyrosine kinase toward the insulin receptor substrate-1 (IRS-1). Phosphorylated IRS-1 then interacts with the p85 alpha subunit of phosphatidylinositol 3-kinase (PI3K), Nck, growth factor receptor-bound protein 2 (GRB2), and Syp, thus branching insulin's signal for both mitogenic and metabolic responses. To determine whether the expression of these proteins is altered in insulin resistance, the levels of these proteins were compared in adipose and liver tissues of nondiabetic mice and obese insulin resistant diabetic KKAy mice. IR and PI3K p85 alpha protein levels were significantly lower in KKAy mice than in control nondiabetic mice, whereas IRS-1 protein levels were not altered. In contrast, the protein levels of GRB2, Nck, Syp, and GLUT-1 were dramatically elevated in KKAy fat, with less striking changes in liver. Treatment of diabetic animals with pioglitazone, an insulin sensitizing antihyperglycemic agent, partially corrected the expression of some of these proteins. Taken together, these findings suggest that the insulin resistant diabetic condition is characterized by changes in expression of insulin signal transduction components that may be associated with altered glucose metabolism. PMID- 7485494 TI - Glucose production in glycogen storage disease I is not associated with increased cycling through hepatic glycogen. AB - Children with glycogen storage disease type I (GSD I) lack the ability to convert glucose 6-phosphate to glucose and yet are able to produce glucose endogenously. To test the hypothesis that the source of this glucose is increased cycling of glucose moieties through hepatic glycogen, six children with GSD I were studied on two occasions during which they received enteral glucose for 6 h at 35 or 50 mumol.kg-1.min-1 along with [6,6-2H2]glucose to measure plasma glucose flux and [1-13C]galactose to label intrahepatic uridyl diphosphate (UDP)-glucose. After 3 h, acetaminophen was given to estimate UDP-glucose flux (reflecting the rate of glycogen synthesis). Mean steady-state plasma glucose concentrations (4.8 +/- 0.2 vs. 5.8 +/- 0.1 mM) and total flux (34.8 +/- 1.7 vs. 47.5 +/- 2.0 mumol.kg-1.min 1) were increased (P < 0.05 or better) on the high-infusion day. Endogenous glucose production was detectable only on the low-infusion day (2.0 +/- 0.5 mumol.kg-1.min-1). UDP-glucose flux was increased (P < 0.05) on the high-infusion day (25.8 +/- 1.6 vs. 34.7 +/- 4.1), ruling out cycling of glucose moieties through glycogen with release of glucose by debrancher enzyme as the source of glucose production. PMID- 7485495 TI - Ionomycin-stimulated phasic myometrial contractions. AB - Ionomycin, a calcium ionophore, facilitates the sustained entry of extracellular calcium; however, in myometrial tissue it stimulates phasic contractions. This study sought to define further this unanticipated effect of ionomycin and to begin to explore the possible mechanism(s) involved. Utilizing rat uterine strips, in vitro isometric contraction studies were performed to determine the effects of ionomycin with and without membrane-permeant inhibitors of cytosolic calcium oscillations. To determine the effects of ionomycin on phospholipase C, qualitative inositol phosphate production studies were performed. The in vitro contraction studies confirmed that ionomycin-stimulated phasic myometrial contractions were potentially dependent on stimulation of phospholipase C, calcium-induced calcium release, and additional calcium influx through dihydropyridine-sensitive membrane calcium channels. The inositol phosphate production studies confirmed that ionomycin stimulated phospholipase C in a dose related fashion to levels comparable to oxytocin. In summary, these observations have confirmed the ability of ionomycin to generate dose-related phasic myometrial contractions through mechanisms potentially involving the phosphatidylinositol-signaling pathway. PMID- 7485496 TI - Prevention of diabetes does not completely prevent insulin secretory defects in the ZDF rat. AB - The rapid insulin secretory pulses that occur in the perfused rat pancreas can be entrained by an oscillatory glucose concentration in pancreata from nondiabetic rats but not from X diabetic Zucker diabetic fatty (ZDF) rats. To investigate whether this defect is present in prediabetic ZDF rats and whether treatment with either pioglitazone or acarbose can prevent or reverse this defect, 39 ZDF and 5 lean ZDF control rats were studied. The ZDF rats were divided into six groups depending on age, form of therapy used, and the time at which pioglitazone was started in relation to the onset of diabetes. The pancreas was isolated and perfused using a sine wave-shaped glucose concentration (mean 7 mM, period 10 min, amplitude 10%). The results, assessed by spectral analysis, revealed that in prediabetic animals and in controls, entrainment of pulsatile insulin secretion was normal. Initiation of pioglitazone therapy in ZDF rats at the time of weaning or before diabetes onset prevented hyperglycemia. However, entrainment was only partially retained. Thus, in these two groups, the spectral power at 10 min was greater than in untreated animals but lower than in prediabetic and control animals. Treatment with acarbose before or with pioglitazone after diabetes onset improved but did not normalize glucose levels, and it did not improve entrainment. The results demonstrate the presence of insulin secretory defects in 13-wk-old ZDF rats in which hyperglycemia was prevented. PMID- 7485497 TI - Tight junctions and the molecular basis for regulation of paracellular permeability. AB - Tight junctions create a regulated paracellular barrier to the movement of water, solutes, and immune cells between both epithelial and endothelial cells. Recent progress has been made in identifying the proteins that create this barrier. The transmembrane protein occludin is an excellent candidate for the sealing protein and is bound on the cytoplasmic membrane surface to the proteins ZO-1 and ZO-2. Functions for ZO-1 and ZO-2 are suggested by their invertebrate homologues, one of which is a tumor suppressor and another is required in epidermal growth factor receptor signaling. Multiple cellular signaling pathways affect assembly and sealing of junctions. Dynamic regulation of perijunctional actin has emerged as a unifying hypothesis for controlling paracellular permeability. Understanding and manipulating permeability will require a more detailed molecular characterization of tight junction proteins and in particular a characterization of how cell signaling regulates their attachment to the perijunctional cytoskeleton. PMID- 7485498 TI - Gender influences sphincter of Oddi response to cholecystokinin in the prairie dog. AB - Although gallstones and disorders of biliary tract motility are both more common in women than men, sphincter of Oddi motility has not previously been compared between the sexes. In this study, cholescintigraphy (under ketamine and diazepam anesthesia) was used to determine gallbladder emptying rate and ejection fraction in response to cholecystokinin (CCK) in eight male and six female prairie dogs fed a nonlithogenic diet. Ten days later, under alpha-chloralose anesthesia, sphincter of Oddi phasic wave activity was monitored for 10-min intervals before (control), during 20 min of CCK infusion, and for 20 min after infusion. Gallbladder emptying rate and ejection fraction and baseline sphincter of Oddi frequency, amplitude, and motility index (= frequency x amplitude) did not differ significantly between the sexes. Sphincter of Oddi phasic wave frequency was increased during CCK infusion in both males and females, but the change in amplitude was significantly greater in females, than males. We conclude that the increased incidence of biliary tract disease in women may be due to altered sphincter of Oddi hormonal response. PMID- 7485499 TI - A novel serine-specific kinase activity associated with exocrine secretory granules. AB - Zymogen secretion from exocrine cells involves an exocytotic process that is highly regulated by the modification of cytoplasmic components at different cellular levels. In the present studies, purified secretory granules were prepared from rabbit gastric chief cells, rat pancreatic acinar cells, and parotid glands to characterize a Mg(2+)-dependent protein kinase activity. In chief cell granules, endogenous pepsinogen, a fortuitous substrate, was phosphorylated at optimal Mg2+ and K+ concentrations of 40 and 50 mM, respectively. Adenosine 3',5'-cyclic monophosphate, Ca2+, and calmodulin had no significant effects on the kinase activity. In contrast, Mn2+ or Zn2+ inhibited the kinase activity. In addition to pepsinogen, the exogenous substrates casein, myelin basic protein, and lysine-rich histone were also phosphorylated by the granule-associated kinase. All substrates were exclusively phosphorylated on serine residues. ATP, but not GTP, served as the donor in the phosphorate transfer reaction. Casein kinase (CK) inhibitors CKI-7 and dibromoribofuransylbenzimidazole at concentrations (10 microM) that significantly inhibited CK activities in the tissue homogenate failed to inhibit the granule associated kinase activity. The kinase activity was localized to the granule membrane and could be removed from the membrane with either 5 mM EDTA or alkaline carbonate extraction. Furthermore, protease digestion sensitivity revealed that the kinase was localized on the cytoplasmic face of the granules. Our results therefore indicate that the secretory granules of exocrine gastric chief cells, pancreatic acini, and parotid acini possess a unique serine-specific protein kinase activity. The cytoplasmic orientation of the kinase activity suggests a possible role in vesicle processing or the exocytotic process. PMID- 7485500 TI - Effect of kinin inhibition in experimental acute pancreatitis. AB - Activation of the endogenous kinin system is a consistent observation in acute pancreatitis and has repeatedly been implicated in the pathophysiology of the disease. We have studied the effect of a potent bradykinin antagonist on the onset and development of acute pancreatitis in four unrelated animal models. Pancreatitis was induced in rats by either supramaximal stimulation with caerulein, intraductal injection of sodium taurocholate, or pancreatic duct ligation with secretin infusion, and in mice by feeding a choline-deficient, ethionine-supplemented diet. The potent, long-acting bradykinin antagonist HOE 140 was administered subcutaneously (0.1 mg/kg every 5 h). Effective kinin inhibition had no effect on pancreatitis-associated mortality, the extent of morphological damage and inflammation, or the intracellular distribution of lysosomal hydrolases. Pancreatic edema was only reduced in caerulein-induced pancreatitis, the only model in which edema formation was paralleled by increased vascular permeability. We conclude that, contrary to previous suggestions, kinins do not play a predominant role in the development of acute pancreatitis. Their participation is strictly limited to vascular events and does not involve the early cell biological alterations in pancreatic acinar cells. PMID- 7485501 TI - Ontogenic regulation of spatial differentiation in the crypt-villus axis of normal and isografted small intestine. AB - Despite rapid proliferation, the mammalian intestinal epithelium maintains precise spatial differentiation from crypt to villus tip and from duodenum to colon. During perinatal life, the rodent gut undergoes a dramatic morphogenesis, resulting in formation of the crypt-villus and duodenal-colonic axes. The ontogeny of regional differences in gene expression in the emerging vertical axis has not been well described. We used the liver fatty acid binding protein (L FABP) and apolipoprotein (apo) AIV genes as markers of neonatal enterocytic differentiation. In situ hybridization analyses revealed that both genes exhibit unique spatial patterns of expression along the jejunal crypt-villus axis during ontogeny, characterized by increased cellular mRNA levels in villus base enterocytes. To examine the requirement for a normal luminal environment to generate these precise patterns of cellular gene expression, we employed intestinal isograft techniques. Fetal intestines were implanted as early as embryonic day 12. Appropriate expression of the apo AIV and L-FABP genes was recapitulated during villus morphogenesis in fetal life. However, spatial patterns of gene expression in the isografted postnatal cryptvillus axis were altered. The preferential accumulation of L-FABP and apo AIV mRNA in villus base enterocytes was never observed in isografts. These results suggest that a "basal" differentiation program is encoded in fetal endoderm and mesenchyme, yet extracellular substances contained in the lumen or extrinsic to the intestine play an important modulatory role in generating spatial differentiation during ontogeny. PMID- 7485502 TI - Effect of short-term starvation on gastric emptying in humans: relationship to oral glucose tolerance. AB - To evaluate the effects of short-term starvation on gastric emptying in normal and obese subjects, the relationship between gastric emptying and oral glucose tolerance, and the mechanisms responsible for the delay in the systemic appearance of oral glucose observed after short-term fasting, we determined the effects of a 4-day fast on 1) gastric emptying and oral glucose tolerance in normal subjects and 2) gastric emptying in obese patients. Gastric emptying of 75 g glucose (320 ml) labeled with 99mTc colloid was measured in 12 healthy volunteers and 11 obese subjects after 12-h and 4-day fasts. In seven other obese subjects, the effect of a 4-day fast on gastric emptying of 320 ml normal saline was quantified. Gastric emptying of glucose was slower after the 4-day than after the overnight fast in both normal (P = 0.02) and obese (P < 0.001) subjects, with no difference between the two groups. In normal subjects, the rate of gastric emptying was related directly to the rise in plasma glucose at 30 min (r = 0.60; P < 0.05) but inversely to the plasma glucose at 180 min (r = -0.64; P < 0.02). In the obese subjects, gastric emptying of saline was not affected by fasting. These observations indicate that 1) gastric emptying of glucose is retarded by a 4-day fast, 2) the changes in gastric emptying reported in obesity may reflect different patterns of prior nutrient intake, and 3) delay in gastric emptying accounts for the slower systemic appearance of glucose after fasting. PMID- 7485503 TI - Rat hepatocytes and Kupffer cells interact to produce interleukin-8 (CINC) in the setting of ethanol. AB - Interleukin-8 is a neutrophil chemoattractant that has been implicated in the pathogenesis of alcoholic hepatitis. The mechanism of ethanol-induced interleukin 8 production in liver is uncertain, although hepatocytes and Kupffer cells have both been proposed as sources of the chemokine. In this study we investigated whether short-term ethanol exposure stimulates production of rat interleukin-8 [cytokine-induced neutrophil chemoattractant (CINC)] by normal rat hepatocytes and Kupffer cells in primary culture. Initial experiments verified that hepatocytes and Kupffer cells produce CINC in response to cytokines or lipopolysaccharide. Ethanol, by contrast, failed to stimulate CINC secretion by either cell type even at concentrations as high as 100 mM. Although ethanol had no direct effect on liver cell CINC production, conditioned medium from ethanol treated hepatocytes induced a threefold rise in CINC production by Kupffer cells. The increase was abrogated when hepatocytes were treated with ethanol and the metabolic inhibitor 4-methylpyrazole. The results suggest that the mechanism of ethanol-induced CINC production is indirect, involving ethanol oxidation and communication between hepatocytes and Kupffer cells. PMID- 7485504 TI - Ischemia-reperfusion increases gastric motility and endothelin-1-induced vasoconstriction. AB - The purpose of our study was to 1) examine the effect of ischemia-reperfusion on gastric vascular resistance and motility, 2) determine whether endothelin-1 (ET 1)-induced vasoconstriction is enhanced after ischemia-reperfusion, and 3) assess the effect of superoxide dismutase (SOD) on these ischemia-reperfusion-induced alterations. These experiments used a mechanically perfused ex vivo gastric segment of chloralose-anesthetized dogs. We first evaluated the effect of varying the duration of total ischemia on reperfusion-induced changes in gastric vascular resistance and motility. In other experiments, responses to ET-1 (10(-10) M) were compared before and after 30-min ischemia and 30-min reperfusion, with saline or SOD (10 U/ml) infused intra-arterially to the stomach during reperfusion. Our results show that 1) after ischemia, vasodilation is seen initially on reperfusion followed by a slowly developing, progressive increase in vascular resistance, 2) the force of gastric contractions was reduced during ischemia but elevated immediately on reperfusion, 3) vasoconstrictor responses to ET-1 are enhanced after ischemia-reperfusion, and 4) SOD reduced the enhanced response to ET-1 and force of contractions. Our findings support the hypothesis that reactive oxygen metabolites contribute to augmented vascular reactivity and hypercontractility after ischemia-reperfusion. PMID- 7485505 TI - Vitamin A-poor lipocytes: a novel desmin-negative lipocyte subpopulation, which can be activated to myofibroblasts. AB - Lipocytes have been classified as vitamin A-storing, desmin-positive cells. In hepatic fibrogenesis, lipocytes transform into myofibroblasts, which express alpha-smooth muscle actin (alpha-SMA) and produce increased amounts of collagen. We isolated a population of vitamin A-poor lipocytes (VAPL) from normal rat liver and examined the morphological and biochemical differences between VAPL and vitamin A-replete lipocytes (VARL). Desmin and alpha-SMA expression were determined by Western blot in quiescent cells and in cells activated by culture on uncoated plastic. Both cell types were alpha-SMA-negative; however, in contrast to VARL, freshly isolated VAPL did not contain desmin. Desmin expression was induced in VAPL on activation. With time in culture, both VAPL and VARL expressed alpha-SMA and produced collagen, indicative of transformation to myofibroblasts. Ferritin receptor expression was demonstrated in cultured VARL after 1 day and in VAPL after 5 days, indicating that this is an early marker of lipocyte activation. After 7 days, VARL and VAPL were indistinguishable in terms of desmin, ferritin receptor expression, and collagen production. This study demonstrates the first isolation and characterization of two distinct quiescent subpopulations of lipocytes from normal rat liver: desmin-negative VAPL and desmin-positive VARL. Both populations of cells can be activated to myofibroblasts, the phenotype associated with hepatic fibrogenesis. PMID- 7485506 TI - Differential expression and regulation of SSTR2 messenger RNA in rat gastric antrum and corpus. AB - Somatostatin modulates both endocrine and exocrine functions in the gastric mucosa, where three of the five cloned somatostatin receptors are present. This study examines changes in somatostatin receptor (SSTR) mRNA abundance during fasting, feeding, and profound acid inhibition with omeprazole. Serum gastrin as well as somatostatin and SSTR mRNA abundances were measured in antrum and corpus. In Northern blots of corpus RNA, the SSTR2 probe hybridized with two previously reported species of mRNA (2.4 and 2.8 kb); in addition, a weak previously unreported 1.6-kb band was detected. In antrum, the 1.6-kb band dominated. Fasting increased antral somatostatin mRNA from 100 +/- 8 to 161 +/- 24% and SSTR mRNA from 100 +/- 10 to 179 +/- 14% (P < 0.05). Omeprazole reduced antral somatostatin mRNA to 34 +/- 4% of control (P < 0.05) and elevated SSTR mRNA to 135 +/- 5% of control (P < 0.01). Omeprazole treatment reduced corpus somatostatin mRNA to 59 +/- 5% (P < 0.05), and elevated SSTR mRNA to 140 +/- 3% of control (P < 0.01). The results therefore indicate that a novel SSTR mRNA subtype exists in the stomach and predominates in the antrum. The abundance of this SSTR mRNA is upregulated by both fasting and achlorhydria; conditions that increase or decrease endogenous antral somatostatin, respectively. PMID- 7485507 TI - Signal transduction by cholera toxin: processing in vesicular compartments does not require acidification. AB - In the polarized human intestinal epithelial cell line T84, signal transduction by cholera toxin (CT) follows a complex series of events in which CT enters the apical endosome and moves through multiple vesicular compartments before it activates adenylate cyclase. As with processing of many other surface ligands, it has been suggested that CT must enter acidic vesicles to exert its downstream effects. To determine if intravesicular pH may regulate signal transduction by CT, we examined the cAMP-dependent Cl- secretory response [short-circuit current (Isc)] to CT in T84 cell monolayers treated with chloroquine (500 microM), methylamine (50 mM), NH4Cl (10 mM), nigericin (4 microM), or bafilomycin A1 (1 microM). Each of these reagents collapsed intravesicular pH gradients as confirmed by accumulation of acridine orange within subcellular compartments of living T84 cells imaged by confocal epifluorescence microscopy. Both acidotropic amines and nigericin inhibited the cAMP-dependent Cl secretory response in T84 cells. However, none of these reagents specifically affected adenylate cyclase itself or coupling of adenylate cyclase with the heterotrimeric guanosinetriphosphatase Gs as judged by the secretory response to the adenosine 3',5'-cyclic monophosphate (cAMP) agonists vasoactive intestinal peptide (VIP), forskolin, or 8-bromo-cAMP. In vitro enzyme-linked immunosorbent assay showed that CT binding to ganglioside GM1 was not dependent on pH between 5.0 and 10. Maximal Isc elicited by apical CT relative to maximal Isc elicited by VIP was not affected by pretreatment with chloroquine, methylamine, NH4Cl, or bafilomycin AI. Nigericin was the only reagent to inhibit CT-induced Isc (5 +/- 2% maximal response to VIP). The data indicate that low intravesicular pH will have little or no effect on CT association with its membrane receptor GM1, or on subsequent processing/signal transduction events. PMID- 7485508 TI - Vagal control of pyloric resistance. AB - Pyloric resistance is probably a major factor regulating gastric emptying of liquids, but its nervous control is unknown. The role of efferent vagal pathways in pyloric resistance was evaluated in 13 anesthetized pigs. Pyloric resistance was assessed by simultaneous recording of gastropyloroduodenal motility and transpyloric flow during gastric emptying of saline. Cervical vagotomy suppressed all antral pressure events, increased the number of pressure events localized at the pylorus, and decreased the frequency of the flow pulses (P < 0.05), without affecting either pyloric resistance or the characteristics of flow pulses. Electrical stimulation of the cervical and the thoracic vagi both decreased pyloric resistance by about 60% and increased the stroke volume of flow pulses (P < 0.05). The reduced pyloric resistance was mainly related to an alteration of the temporal relationship between flow pulses and pyloric pressure events. These results indicate that vagal efferents could provide inhibitory inputs to pyloric resistance. A reduction in pyloric resistance contributes to the increased flow rate observed during vagal stimulation. PMID- 7485510 TI - Influence of breathing pattern on the esophagogastric junction pressure and esophageal transit. AB - The esophagogastric junction (EGJ) is guarded by two sphincters, a smooth muscle lower esophageal sphincter (LES) and a skeletal muscle crural diaphragm. The LES relaxes in response to a swallow but the crural diaphragm does not. Since contraction of the crural diaphragm is affected by the pattern of breathing, the latter may also influence the EGJ pressure and swallow-induced EGJ relaxation. Our aims were to study the effects of alterations of the breathing pattern on the EGJ pressure, swallow-induced EGJ relaxation, and esophageal transport of liquid bolus. Manometric, electromyographic, and videofluoroscopic studies were performed in 12 healthy subjects. The subjects were trained in two types of breathing patterns, hyperventilation and partial expiration, using the visual biofeedback from their own respiratory waveform. Hyperventilation increased the frequency of inspiratory pressure oscillations at the EGJ without affecting the end-expiratory EGJ pressure. Partial expiration resulted in an increase in the end-expiratory EGJ pressure. Swallow-induced relaxation at the EGJ was markedly reduced during partial expiration. An inspiration during a swallow (control breathing and hyperventilation) caused transient interruption of flow across the EGJ. Partial expiration resulted in cessation of the flow across the EGJ, failure of esophageal peristalsis to traverse the entire length of the esophagus, increased esophageal transit time, and incomplete esophageal clearance of a liquid bolus.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7485509 TI - Evidence for a low Km transporter for non-transferrin-bound iron in isolated rat hepatocytes. AB - Non-transferrin-bound iron (NTBI) plays an important role in the hepatocellular injury induced by iron overload. However, the mechanism responsible for NTBI uptake into hepatocytes remains poorly defined. The purpose of this study was to define the kinetics of NTBI uptake by isolated rat hepatocytes and to characterize the uptake process. NTBI uptake was time and temperature dependent, exhibited a Michaelis-Menten constant (Km) value of 1.25 microM and maximum uptake of 241 pmol.10(6) cells-1.min-1, and 55Fe was incorporated in part into intracellular ferritin. Uptake was Ca2+ dependent, exhibiting 15 and 80% of maximal uptake in the presence of 0.6 and 0.75 mM CaCl2, respectively. The putative NTBI transporter was highly specific; divalent (Zn2+, Mn2+, Cd2+, and Co2+) or trivalent (La3+) cations did not inhibit Fe3+ uptake. Reduction from Fe3+ to Fe2+ was not essential for uptake or the process occurred deep within the membrane bilayer, since the Fe2+ chelator ferrozine did not influence 55Fe uptake. These data provide evidence for a low Km plasma membrane transporter for NTBI, which should be functional at physiological serum concentrations and saturated in iron-overload diseases, such as hemochromatosis. PMID- 7485512 TI - L-glutamine and L-asparagine stimulate ODC activity and proliferation in a porcine jejunal enterocyte line. AB - We studied the effect of L-glutamine (Gln), the principal intestinal fuel, on proliferation of a porcine jejunal cell line, IPEC-J2. In cells synchronized by serum deprivation for 4 h, Gln stimulated ornithine decarboxylase (ODC; EC 4.1.1.17) in a dose- and time-dependent manner, with maximal effects at 10 mM in 3 h (P < 0.01). Similar effects were seen for the structurally related amino acid L-asparagine and serum. The Gln effect on ODC was specific, as isosmolar mannitol, glucose, methyl-beta-D-glucoside, L-phenylalanine, ammonia, and aminoisobutyric acid were ineffective. The alanine aminotransferase inhibitor aminooxyacetate (AO) inhibited the ODC stimulation by Gln in a dose-dependent manner (half-maximal inhibitory concentration = 0.5 mM). AO was not toxic to cells, as determined by propidium iodide uptake into nuclei. In addition, Gln stimulated a twofold increase of cellular 24-h [3H]thymidine incorporation above rates of control cells bathed in standard media (P < 0.01); this effect was also blocked by AO. Gln and phorbol 12-myristate 13-acetate stimulated ODC in a synergistic manner. The Na+/H+ exchange inhibitor methylisobutyl amiloride blocked the enhancement of ODC by Gln. Gln also induced the mRNA of the immediate early gene c-jun. Gln stimulates proliferation in a porcine jejunal cell line through a mechanism requiring transamination and intact Na+/H+ exchange. This stimulation of enterocyte proliferation by Gln suggests that therapeutic Gln administration could facilitate epithelial recovery in the injured small intestine. PMID- 7485511 TI - Effect of 17-norleucine-VIP on gastroduodenal motility relative to serum VIP concentration and blockade of NOS. AB - Mechanical and electrical activity in the antrum, pylorus, and duodenum was evaluated in the conscious dog, instrumented with seven strain gauges and five platinum electrodes. 17-Norleucine-vasoactive intestinal peptide (17-N-Leu-VIP) or 17-N-Leu-VIP plus NG-nitro-L-arginine methyl ester (L-NAME) was injected intra arterially close to the pylorus to identify influences of nitric oxide (NO) on effects of VIP. VIP concentration was measured by radioimmunoassay in serum samples collected from the cubital and portal veins before and up to 2 h after VIP injection. VIP (0.004-0.006 mg.kg-1.10 min-1) abolished phasic contractions in the interdigestive state for 16.8 min and in the digestive state for 14.4 min, whereas whole serum VIP concentration rose above 42.4 +/- 13 pmol/l. Administration of L-NAME did not significantly influence the effects of VIP. Aftereffects of VIP, consisting of a reduced motility index, lasted 33 +/- 10.6 min in the interdigestive state and 44.5 +/- 42 min in the digestive state. This VIP aftereffect in the interdigestive state was shortened in time by the addition of L-NAME. The results overall suggest that NO release is a factor only in the aftereffects of VIP. PMID- 7485513 TI - Metabolism and influence of gastrin-52 on gastric acid secretion in humans. AB - It has been shown recently that the two largest alpha-carboxyamidated progastrin products are gastrin-71 and gastrin-52. Human gastrin-52 has now been synthesized, and the effect on gastric acid secretion and elimination from plasma was examined and compared with gastrin-17 in 12 normal subjects. The peptides were infused separately in four consecutive doses; the maximum response of gastrin-17 and gastrin-52 was 25.2 +/- 2.8 and 22.2 +/- 2.8 mmol H+/50 min, respectively (P < 0.01). This difference in efficacy was presumably related to nonequilibrium of gastrin-52 between plasma and receptor. The elimination of gastrin-17 was monoexponential with a half-life of 4.7 +/- 0.3 min; clearance and apparent volume of distribution were 16.7 +/- 1.5 ml.kg-1.min-1 and 106.0 +/- 9.2 ml/kg, respectively. The elimination of gastrin-52 was biexponential, the half lives were 4.9 +/- 0.7 and 49.9 +/- 4.2 min, and clearance and apparent volume of distribution were 1.9 +/- 0.2 ml.kg-1.min-1 and 106.3 +/- 10.1 ml/kg, respectively. Gel chromatography of plasma samples drawn during infusion of gastrin-52 revealed that most of the immunoreactivity eluted in the position of the intact peptide. Small peaks in the positions of gastrin-34 and the NH2 terminal pentapeptide fragment of gastrin-52 indicate that a minor part of gastrin-52 is degraded to smaller peptides in vivo. It is concluded that gastrin 52 is bioactive with an efficacy close to or similar to that of gastrin-17. A minor fraction of gastrin-52 undergoes postsecretory cleavage either in plasma or after capillary transit. PMID- 7485514 TI - Nitric oxide modulates a calcium-activated potassium current in muscle cells from opossum esophagus. AB - Nitric oxide mediates nerve-induced hyperpolarization of circular smooth muscle of the esophagus. Two mechanisms are proposed to explain this hyperpolarization: an increase in K+ current or a decrease in Cl- current. These studies test the hypothesis that nitric oxide increases a K+ current in esophageal smooth muscle. Three outward K+ currents are present in circular smooth muscle cells from the opossum esophagus. One current is a Ca(2+)-activated K+ current (IKCa2+). This current is inhibited by charybdotoxin. Whole cell currents were recorded from isolated opossum esophageal smooth muscle cells using the whole cell patch-clamp technique. These studies showed that IKCa2+ is activated at potentials more positive than -30 mV. Bath application of S-nitroso-L-cysteine increased IKCa2+ by 50% above control levels throughout the entire activation range of potentials. The enhanced current was reversible on washout. Either charybdotoxin, an inhibitor of IKCa2+, or (R)-p-8-(4-chloropenylthio)-guanosine 3',5'-cyclic monophosphorothioate, an inhibitor of protein kinase G, antagonized the increase in outward current induced by S-nitroso-L-cysteine. These data suggest that nitric oxide activates IKCa2+ via the guanosine 3',5'-cyclic monophosphate protein kinase G signal transduction pathway. PMID- 7485515 TI - Transcription factors and pattern formation in the developing lung. AB - During development of the respiratory tract embryonic cells are instructed to organize themselves along an axis and differentiate, such that proximal structures (trachea) greatly differ from those in distal alveoli. Pattern formation relates to this process of organization, and it is believed to be transcriptionally regulated in many developmental systems. Although the lung is the site of expression of many transcription factors, such as Hox, retinoid receptors, hepatocyte nuclear factors, and myc, among others, little information is available on how they influence lung pattern. Functional studies so far have directly implicated the product of the protooncogene N-myc and the retinoic acid receptors as transcriptional regulators of lung patterning, and it is likely that tissue-specific homeobox genes, such as the thyroid transcription factor-1, play an important role in distal lung formation. This review describes several aspects of transcription factors possibly involved in lung patterning, including structure, spatial distribution, and their putative functions. PMID- 7485516 TI - Corticosteroid inhibition of macrophage inflammatory protein-1 alpha in human monocytes and alveolar macrophages. AB - One of the major inducible cytokines secreted by mononuclear phagocytes is macrophage inflammatory protein 1 (MIP-1), which consists of two homologous polypeptides, MIP-1 alpha and MIP-1 beta. MIP-1 alpha possesses chemotactic and stimulatory activities for lymphocytes, eosinophils, and monocytes and may play a role in various pulmonary inflammatory conditions. We investigated the expression and release of MIP-1 alpha from human peripheral blood monocytes (PBM) and alveolar macrophages (AM) after stimulation with lipopolysaccharide (LPS), interleukin-1 beta (IL-1 beta), tumor necrosis factor-alpha, and interferon-gamma and the inhibitory effects of corticosteroids. LPS and IL-1 beta only enhanced MIP-1 alpha mRNA and protein in a dose- and time-dependent fashion. Dexamethasone (10(-9) to 10(-4) M) inhibited the basal and induced production and expression of MIP-1 alpha. In PBM, dexamethasone (10(-6) M) reduced LPS- and IL-1 beta stimulated production of MIP-1 alpha protein by 50 and 63%, respectively, maximally at 24 h, whereas the inhibition of mRNA expression occurred maximally at 4 h. Similar trends were observed for AM. MIP-1 alpha mRNA decay was only slightly decreased in the presence of dexamethasone. Inhibition of LPS-induced MIP-1 alpha mRNA by dexamethasone was attenuated by the protein synthesis inhibitor cycloheximide, indicating the involvement of a protein intermediate. Corticosteroids are a potent inhibitor of IL-1 beta- and LPS-induced expression of MIP-1 alpha through mechanisms involving mainly inhibition of transcription and to a minor degree by reducing mRNA stability. Corticosteroids may be effective anti-inflammatory agents by preventing the expression of chemokines such as MIP-1 alpha. PMID- 7485518 TI - Retinoids, retinoic acid receptors, and cytoplasmic retinoid binding proteins in perinatal rat lung fibroblasts. AB - The early postnatal life of most mammals marks a period of extensive enlargement of the alveolar surface area and increase in the elastin content of the lung. The factors that regulate the onset and abatement of this burst of elastin synthesis have not been identified. Previous studies of lipid-laden rat pulmonary interstitial fibroblasts (LIF) have shown that their elastin synthesis is increased in vitro by retinoic acid (RA). We hypothesized that temporal changes in the endogenous RA content of LIF may correlate with changes in elastin synthesis by these cells. LIF were isolated from the lungs of rats at gestational day 19 and postnatal days 2, 4, 8, and 12 and their retinoid contents were quantitated. Retinyl esters were highest at gestational day 19 (2.9 +/- 0.6 pmol/10(6) cells, means +/- SE) and decreased to 1.6 +/- 0.2 pmol by postnatal day 2 (P < 0.05). This decrease in retinyl esters was accompanied by an increase in retinol from 0.4 +/- 0.1 to 2.0 +/- 0.6 pmol/10(6) cells (P < 0.05). RA increased in LIF from 0.07 +/- 0.04 pmol/10(6) cells at gestational day 19 to 0.29 +/- 0.05 pmol/10(6) cells at postnatal day 2 (P < 0.05) and increased in whole lung tissue from 0.07 +/- 0.04 to 0.29 +/- 0.05 nmol/g, over the same interval. The increase in RA content was accompanied by an increase in RA receptor (RAR)-beta and -gamma mRNAs. The steady-state mRNA level of cellular retinol binding protein (CRBP) was high in LIF, relative to whole lung tissue at day 2. Cellular RA binding protein (CRABP) mRNA rose fourfold from day 2 to day 8 and then fell by day 12. In summary, RA, RAR, and CRBP mRNA in LIF are highest before the period of maximal elastin synthesis, which occurs at postnatal days 8 and 12. These findings are consistent with the hypothesis that endogenous RA could contribute to the postnatal increase in elastin production by pulmonary fibroblasts. PMID- 7485517 TI - Cellular uptake of albumin from lungs of anesthetized rabbits. AB - Resolution of alveolar edema depends on clearance of serum protein, as well as liquid from the alveolar space. Protein clearance is slower than liquid clearance and may take days to weeks. Our earlier studies presented evidence for the importance of paracellular removal of soluble protein from the air spaces. However, long-term protein clearance may also depend on uptake by alveolar epithelial cells or macrophages. This study examined cellular uptake of soluble human albumin and insoluble colloidal gold-albumin from the lungs of anesthetized rabbits. Native albumin was endocytosed by both alveolar type I and type II cells and appeared in vesicles and endosomes. Neither cell type took up colloidal gold albumin over periods as long as 8 h. Alveolar macrophages took up native albumin and colloidal gold-albumin to a greater extent and more rapidly than alveolar epithelial cells. The tracer proteins were found in vesicles, endosomes, and phagolysosomes. Similarly, cultured alveolar macrophages took up native albumin more rapidly than cultured type II cells. Thus macrophages may be important in clearing precipitated protein from the air spaces, and they may have a role in completing the clearance of soluble protein. The potential for transepithelial transport of soluble alveolar protein exists, but based on this work and our prior studies, it appears to be a low-capacity pathway. PMID- 7485519 TI - Glutathione S-transferases in tracheobronchial epithelium. AB - The purpose of this study is to characterize glutathione S-transferase (GST) gene expression in airway epithelium both in vivo and in vitro. Immunohistochemical staining of nonhuman primate lungs of well-controlled healthy animals reveals the presence of alpha- and pi-class GST isoenzymes in ciliated bronchial epithelium. The stain of mu-GST antibody is either very low or absent in some of these monkey lungs. We observed that primary tracheobronchial epithelial (TBE) cells isolated from human and monkey pulmonary tissues maintain a relatively high level of GST enzymatic activity in culture, compared with various immortalized human TBE cell lines and other nonpulmonary cell lines. Northern blot analysis demonstrated the presence of mu-, pi-, and microsomal-GST messages but not the alpha-class message in cultures of primary TBE cells as well as in various human TBE cell lines. The expression of mu- and pi-class GST genes can be further regulated in culture by various environmental factors; however, most of these regulating factors are associated with TBE cell differentiation in culture. For instance, vitamin A treatment, which was shown to enhance mucous cell differentiation in vitro, stimulated the message levels of mu- and pi-class GST. Furthermore, plating cells on collagen gel substrata, which also enhanced mucous cell differentiation in culture, instead of plastic culture surface, enhanced total GST enzymatic activity by eightfold, and this enhancement is related to an increase in the expression of the pi-class GST gene. These results demonstrated that GST genes are differentially expressed and regulated by various environmental factors in primary TBE cells and various cell lines, and the regulation is correlated to the mucous cell differentiation in culture. PMID- 7485520 TI - Tenascin in rat lung development: in situ localization and cellular sources. AB - Tenascin (TN) is a hexameric extracellular matrix glycoprotein that may play an important role during lung development. TN protein is temporally and spatially restricted during lung organogenesis. The temporo-spatial and cellular expression of TN mRNA in lung remains unclear. Localization of message expression of TN in rat lung tissue was first investigated by using in situ hybridization performed with an antisense RNA probe. TN mRNA was present primarily within the mesenchyme of day 16 gestational age fetal rat lung tissue, whereas immunoreactive TN protein was found along the basement membrane. In postnatal day 3 rat lung tissue, TN mRNA was detected along alveolar septal walls and was concentrated at secondary septal tips. Expression of TN message was consistent with localization of immunoreactive TN protein. Accumulation of TN mRNA in alveolar septal tips suggests that mesenchyme may be the major source of TN mRNA. To investigate the cellular source of TN in rat lung, we studied the expression of TN in cultured rat lung fibroblasts, endothelial cells, and alveolar epithelial cells. Two TN isoforms having molecular mass of 230 and 180 kDa were in conditioned medium and in cellular extracts of lung fibroblasts and endothelial cells. TN was secreted and deposited in the extracellular matrix closely associated with the surface of lung fibroblasts and endothelial cells. Lung alveolar epithelial cells showed undetectable or barely detectable amounts of TN. These studies demonstrated that TN isoforms are expressed not only by lung fibroblasts but also by lung endothelial cells. The unique spatial localization of TN mRNA during lung development and expression of TN by different lung cell types suggested TN may be involved in matrix organization and cell-cell interactions during lung development. PMID- 7485521 TI - Phosphocholine reverses inhibition of pulmonary surfactant adsorption caused by C reactive protein. AB - The influence of the acute inflammatory phase protein human C-reactive protein (CRP) on the adsorption of porcine pulmonary surfactant from a subphase into an air-water interface has been investigated. CRP was shown to detract from the ability of surfactant to rapidly adsorb to the air-water interface at a molar ratio of 0.03:1 (protein:phospholipid) (weight ratio, 0.5:1). On a weight basis, CRP was found to be more effective than fibrinogen at reducing the adsorption rate of surfactant. The effect of CRP required the presence of calcium and was reversed by the addition of phosphocholine in a concentration-dependent manner. The inhibition of surfactant adsorption by CRP was effectively eliminated by the addition of phosphocholine at a molar ratio of 300:1 (phosphocholine:CRP), but it was not diminished by the addition of identical molar ratios of o phosphoethanolamine or DL-alpha-glycerophosphate at the same molar ratios. These data suggest that the potent inhibition of surfactant adsorption by CRP is primarily a result of a specific interaction between CRP and the phosphocholine headgroup of surfactant lipids in the subphase and that it can be reversed by the water-soluble CRP ligand, phosphocholine. PMID- 7485522 TI - Enhanced proliferation and IL-2 secretion by lung lymphocytes from HIV-infected subjects. AB - Human immunodeficiency virus (HIV)-positive patients frequently develop a CD3+/CD8+ cytotoxic T cell lymphocytic alveolitis. This could occur through in situ expansion of lung lymphocytes. We evaluated lung and blood lymphocyte proliferation in asymptomatic HIV-infected individuals by measuring spontaneous and cytokine-induced tritiated thymidine incorporation. Interleukin (IL)-2 and IL 4 secretion was determined with the use of enzyme-linked immunosorbent assay, Western blotting, and immunoprecipitation techniques. Spontaneous proliferation by lung lymphocytes from HIV-positive patients was significantly greater than that of normal volunteers. Proliferation was confined to the CD8+ lymphocyte subset. Over time, spontaneous proliferation declined unless autologous alveolar macrophages (AM) were added, suggesting AM were providing additional stimulatory signals to lung lymphocytes. Lung and blood lymphocytes proliferated in response to IL-2 but not IL-4. Lymphocytes in HIV-infected lung spontaneously produced and secreted more IL-2 than either normal lung lymphocytes or autologous blood lymphocytes. IL-4 production was not detectable in either group. These findings support the hypothesis that lymphocytic alveolitis in asymptomatic HIV-positive patients results from IL-2-dependent in situ proliferation of CD3+/CD8+ cytotoxic T cells. PMID- 7485523 TI - Influence of PLC and MLCK inhibitors and the role of L-calcium channels in the cat pulmonary vascular bed. AB - The effects of U-73122, a phospholipase C (PLC) inhibitor, on pressor responses to angiotensin II (ANG II), norepinephrine (NE), serotonin (5-HT), BAY K 8644, and the thromboxane A2 (TxA2) mimic, U-46619, were studied in the pulmonary vascular bed of the intact-chest cat. Under conditions of constant lobar blood flow, injections of ANG II, NE, 5-HT, U-46619, and the calcium channel opener, BAY K 8644, into the lobar arterial perfusion circuit caused dose-related increases in lobar arterial pressure, which were reproducible with respect to time. Infusion of U-73122, a PLC inhibitor, into the perfused lobar artery at 10 100 micrograms/kg for 10 min significantly reduced responses to ANG II, serotonin, and NE; however, U-73122 did not alter responses to BAY K 8644 or to U 46619. In a separate series of animals, the effects of the myosin light chain kinase inhibitor, KT-5926, were investigated, and after infusion of KT-5926 into the perfused lobar artery at 1-2 micrograms/kg for 10 min, responses to ANG II, NE, 5-HT, BAY K 8644, and U-46619 were reduced significantly. In a final series of experiments, the effects of the L-type calcium channel blocker, nicardipine, were investigated, and infusion of the L-type calcium channel blocker into the perfused lobar artery at 0.5-1 microgram/kg for 10 min reduced responses to ANG II, BAY K 8644, and NE. However, nicardipine did not alter pressor responses to 5 HT or the TxA2 mimic, U-46619.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7485524 TI - Heparin and PGE2 inhibit DNA synthesis in human airway smooth muscle cells in culture. AB - An increase in the bulk of the airway smooth muscle is a characteristic of asthma. Much of the research investigating the mechanisms of this increase in muscle has focused on mediators that are mitogenic for smooth muscle, while relatively few studies have focused on mediators inhibiting mitogenesis. In this study we have examined the effects of two mediators proposed as regulators of smooth muscle proliferation, namely heparin and prostaglandin (PG) E2, on human airway smooth muscle cells in culture stimulated with 1, 2.5, 5, and 10% fetal bovine serum (FBS) and platelet-derived growth factor AB (PDGF), 50 ng/ml. PGE2 had a biphasic effect on DNA synthesis in the presence of 1% FBS, with 10(-6) M causing inhibition and 10(-7) M causing an increase in DNA synthesis. PGE2 caused inhibition of DNA synthesis in the presence of 2.5, 5, and 10% FBS. Heparin (10 and 100 U/ml) caused an inhibition of DNA synthesis induced by 1% FBS, while 100 U/ml inhibited DNA synthesis induced by 5 and 10% FBS. PGE2 (10(-8), 10(-7), and 10(-6) M) inhibited the DNA synthesis induced by PDGF, while heparin (1, 10, and 100 U/ml) had no effect. These results indicate that both PGE2 and heparin may have a role in the control of human airway smooth muscle cell growth. PMID- 7485525 TI - Regulation of fluid-phase endocytosis in alveolar macrophages. AB - We investigated whether fluid-phase endocytosis in rabbit alveolar macrophages (AM) was regulated by alterations in intracellular adenosine 3',5'-cyclic monophosphate (cAMP). Suspensions of freshly isolated AM were incubated with anionic dextrans (mol mass = 10 kDa), coupled to fluorescein isothiocyanate (FITC), at either 37 or 4 degrees C. There was a rapid increase in AM-associated fluorescence, quantified by laser flow-cytometry and video microscopy during the first hour of incubation at 37 degrees C, which was directly proportional to the amount of tracer present in the medium. In contrast, at 4 degrees C, AM fluorescence was similar to autofluorescence. Incubation of AM with forskolin (50 microM) or 3-isobutyl-1-methyl xanthine (IBMX; 0.1 mM) increased their cAMP content by 67 +/- 2 and 52 +/- 5% (mean +/- SE; n = 4) and decreased FITC-dextran uptake by 29 +/- 4 and 31 +/- 4% (n = 3). On the other hand, incubation of AM with 0.5 mM IBMX inhibited FITC-dextran uptake by 62 +/- 4% (n = 3), without any further increase in cAMP. Incubation of AM with 0.4 mM 8-(4-chlorophenylthio) adenosine 3',5'-cyclic monophosphate (CPT-cAMP), a cell-permeable analogue of cAMP, decreased FITC-dextran uptake by 48 +/- 5% (n = 6). Pulse-chase experiments showed that the rate of FITC-dextran exocytosis was not affected by cAMP. We concluded that fluid-phase endocytosis in rabbit AM is regulated by cAMP and by an additional, cAMP-independent mechanism of IBMX. PMID- 7485526 TI - Neutrophils enhance removal of ozone-injured alveolar epithelial cells in vitro. AB - After acute exposure to oxidant gases in vivo, migration and accumulation of inflammatory cells in pulmonary epithelium coincides with epithelial cell necrosis. The present study was designed to test quantitatively the hypothesis that quiescent neutrophils enhance the removal of oxidant-injured pulmonary epithelial cells after exposure to ozone in vitro. Primary isolated rat alveolar type II cells were cultured as monolayers, using serum-free medium. After exposure to 0.1-0.5 ppm ozone for 0.5 h, apical sides of monolayers were administered either fresh nutrient medium only or medium containing quiescent human neutrophils. Monolayer bioelectric properties and cellular uptake of vital dye were recorded from 5 to 48 h after ozone exposure. Ozone dose-dependent increases in monolayer permeability were associated with proportionally higher numbers of injured epithelial cells. However, the direction and magnitude of neutrophil effects on monolayer permeability after ozone exposure were dependent on ozone concentration. Furthermore, neutrophil-treated monolayers exposed to 0.1 ppm ozone had significantly fewer attached cells positive for uptake of vital dye relative to monolayers exposed to the low level of ozone only; this effect was ablated with increasing ozone concentration. These data suggest that at high levels of ozone neutrophils may exacerbate injury to oxidant-impaired epithelial cells, whereas the presence of neutrophils after exposure to ambient concentrations of ozone may expedite the restoration of epithelial barrier function. We conclude that, by enhancing the removal of injured cells, neutrophils may facilitate the repair of centriacinar epithelium after ozone exposure in vivo. PMID- 7485527 TI - Regulation of cytokeratin expression in rat lung alveolar epithelial cells in vitro. AB - Type II pneumocytes are the stem cells for the alveolar epithelium, proliferating and differentiating into type I cells during lung growth or after injury. The pattern of cytokeratin expression by type II cells in vitro has been linked to the state of differentiation of these cells. In particular, cytokeratin 19 expression has been associated with the type II cell phenotype. We now examine the roles of cell shape and cell-cell interactions in the regulation of cytokeratin expression by rat type II cells in vitro. Type II cell spreading and intercellular contacts were modulated by seeding cultures at high or low density (3.5 or 0.5 x 10(5) cells/cm2). When cultured at high density, cells demonstrated increased cytokeratin 19 protein synthesis and diminished cytokeratin 18 protein synthesis compared with highly spread cells at low density. This effect was a reflection of changes in the abundance of mRNAs for the individual cytokeratins. Alveolar epithelial cells at high density also formed extensive desmosomes between the cells. Desmosome formation was significantly decreased when cells were seeded at low density or in reduced (0.05 mM) calcium medium. Cytokeratin 19 mRNA and protein expression were also significantly decreased when desmosome formation was inhibited in reduced calcium medium, while calcium content of the medium had little effect on cytokeratin 18. These studies suggest that type II cell expression of cytokeratin 19, a differentiation-related cytokeratin, is regulated by factors influencing cell shape and intercellular contacts between epithelial cells. They further suggest that cell-cell interactions between epithelial cells may play a role in the modulation of epithelial cell phenotype. PMID- 7485528 TI - Combined effects of nitric oxide and hyperoxia on surfactant function and pulmonary inflammation. AB - NO and its derivative ONOO- are potent free radicals that can cause cell damage, especially in the presence of O2. To determine the potential pulmonary toxicities of nitric oxide (NO) and peroxynitrite (ONOO-) in vitro, Survanta (2.5 mg/ml) was exposed to ONOO- (0.3-8 mM) in the presence of two different buffering systems (N 2-hydroxyethylpiperazine-N'-2-ethanesulfonic acid and phosphate buffer) and minimum surface tension (MST) was determined with an oscillating bubble surfactometer. Significant increases in MST were seen only with exposure to 8 mM ONOO-, indicating that in vitro, high concentrations of ONOO- can inhibit natural surfactant function. The in vivo effects of NO and hyperoxia were then studied in four groups of newborn piglets ventilated for 48 h with 21% O2, 100% O2, 21% O2 and 100 ppm NO, or with 90% O2 and 100 ppm NO. Five animals served as an untreated control group. Bronchoalveolar lavage fluid (BAL) obtained at 48 h was subjected to centrifugation and the surfactant pellet was reconstituted to 5 mg phospholipid/ml. Significant increases in MST were seen in surfactant from piglets ventilated with NO and 90% O2, compared with either untreated controls or piglets ventilated with 21% O2 for 48 h (P < 0.05, analysis of variance). Significant increases in neutrophil chemotactic activity (NCA) of BAL were also found in the NO and O2 group (P < 0.05), with significant positive interaction between NO and O2 found (P < 0.01).(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7485529 TI - TNF-alpha induces peroxynitrite-mediated depletion of lung endothelial glutathione via protein kinase C. AB - We tested the hypothesis that tumor necrosis factor-alpha (TNF) induces a peroxynitrite (ONOO-)-mediated depletion of glutathione via a protein kinase C (PKC)-dependent mechanism in pulmonary artery endothelial monolayers (PAEM). PAEM were incubated with TNF (1,000 U/ml) for 6 and 18 h. The PAEM were assayed for ONOO(-)-dependent changes in the concentration of luminol, free glutathione [Gfree; i.e., reduced glutathione and oxidized glutathione (GSSG)] and GSSG. TNF treatment decreased luminol and Gfree, and increased GSSG and GSSG/Gfree, compared with treatment with control media. The TNF-induced effects were prevented by co-incubation with the nitric oxide synthase inhibitors NG monomethyl-L-arginine (1 mM), NG-nitro-L-arginine methyl ester (1 mM), or NG nitro-L-arginine (1 mM). In addition, the TNF-induced effects were prevented by superoxide dismutase (10 U/ml), which removes O2-, and by urate (0.5 mM) and L cysteine (3 mM), putative scavengers of ONOO-. The treatment of PAEM with the PKC activator phorbol 12-myristate 13-acetate (PMA, 1 microM) induced similar alterations in luminol and glutathione as TNF. TNF and PMA induced a protein of similar molecular weight (approximately 90 kDa) in the focal contact-rich fraction of PAEM lysate. TNF- and PMA-induced effects were prevented with the specific PKC inhibitor calphostin C (1 microM). The data indicate that TNF induced PKC activation mediates ONOO- generation, which results in the oxidation and depletion of glutathione in PAEM. PMID- 7485531 TI - Selective A1 adenosine receptor antagonism augments beta-adrenergic-induced renin release in vivo. AB - This study determines, in vivo, whether endogenous adenosine/A1 receptor interactions at juxtaglomerular cells restrain the release of renin induced by receptor-mediated activation of the adenosine 3',5'-cyclic monophosphate pathway and whether endogenous adenosine/A2 receptor interactions diminish this restraining response. The following four pharmacological probes were employed: 1) 1,3-dipropyl-8-cyclopentylxanthine (DPCPX) and 2) FK-453, both selective A1 receptor antagonists; 3) FR-113452, a nearly inactive enantiomer of FK-453; and 4) KF-17837, a selective A2 receptor blocker. Adult Sprague-Dawley rats were prepared (adrenalectomized, renal denervated, uninephrectomized, and treated with indomethacin, aldosterone, and hydrocortisone) to minimize endogenous stimulation of renin release and received either vehicle (control group) or one of the four drugs. Intrarenal infusions of isoproterenol (3, 30, and 100 ng.kg-1.min-1) caused dose-related increases in plasma renin activity (PRA). This PRA response was significantly augmented in the groups receiving DPCPX (P = 0.0010) or FK-453 (P = 0.0001) but was not altered in the groups treated with FR-113452 (P = 0.3422) or KF-17837 (P = 0.2155). Systemic and renal hemodynamics and renal electrolyte excretions were monitored and could not account for the PRA augmentation caused by the A1 antagonists. This study clearly demonstrates that endogenous adenosine acts on the A1 receptor to restrain the renin release induced by activation of intrarenal beta-adrenoceptors and is not counteracted by endogenous activation of the A2 receptor. PMID- 7485530 TI - Regional and segmental localization of AE2 anion exchanger mRNA and protein in rat kidney. AB - Chloride/base exchange activity has been detected in every mammalian nephron segment in which it has been sought. However, in contrast to the Cl-/HCO3- exchanger AE1 in type A intercalated cells, localization of AE2 within the kidney has not been reported. We therefore studied AE2 expression in rat kidney. AE2 mRNA was present in cortex, outer medulla, and inner medulla. Semiquantitative polymerase chain reaction of cDNA from microdissected tubules revealed AE2 cDNA levels as follows [copies of cDNA derived per mm tubule (+/- SE)]: proximal convoluted tubule, 688 +/- 161; proximal straight tubule, 652 +/- 189; medullary thick ascending limb, 1,378 +/- 226; cortical thick ascending limb, 741 +/- 24; cortical collecting duct, 909 +/- 71; and outer medullary collecting duct, 579 +/ 132. AE2 cDNA was also amplified in thin limbs and in inner medullary collecting duct. AE2 polypeptide was detected in all kidney regions. AE2 mRNA and protein were also detected in several renal cell lines. The data are compatible with the postulated roles of AE2 in maintenance of intracellular pH and chloride concentration and with its possible participation in transepithelial transport. PMID- 7485534 TI - Regulation of H(+)-K(+)-ATPase expression in kidney. AB - It is now widely accepted that proton secretion by the collecting duct is mediated, in part, by an H(+)-K(+)-adenosinetriphosphatase (H(+)-K(+)-ATPase). Controversy persists regarding which H(+)-K(+)-ATPase isoform is expressed in kidney. Several laboratories have reported preliminarily the amplification from kidney of stomach and/or colon-identical products using gastric or colonic specific primers in the polymerase chain reaction (PCR). We have developed highly specific probes for the catalytic subunit using reverse transcriptase-PCR with gastric- or colonic-specific primers. The resulting cDNAs were verified by sequencing and were then used in Northern analysis of whole kidney total RNA obtained from one of the following three groups of rats: 1) controls, 2) chronic hypokalemia, or 3) chronic metabolic acidosis. Probes for both the colonic and gastric alpha-subunit H(+)-K(+)-ATPase isoforms hybridized to whole kidney total RNA derived from potassium-replete control rats. A marked elevation of colonic mRNA abundance, but not gastric message, was observed in response to chronic hypokalemia induced by dietary potassium deprivation. Elevation of either gastric or colonic mRNA was not observed with chronic metabolic acidosis. Under the conditions of the present study, it appears that the mRNA encoding the colonic alpha-isoform of the H(+)-K(+)-ATPase in kidney is upregulated by chronic hypokalemia but not by chronic metabolic acidosis. The observation that the gastric H(+)-K(+)-ATPase alpha-isoform does not appear to be regulated in either condition suggests that this isoform is expressed constitutively in kidney. PMID- 7485532 TI - Restoration of urine concentrating ability and accumulation of medullary osmolytes after chronic diuresis. AB - Restoration of urine osmolality (Uosm) and medullary osmolyte contents after chronic diuresis was studied in rats infused for 6 days with furosemide and subsequently given the vasopressin analogue, 1-desamino-8-D-arginine vasopressin (DDAVP). Papillary tip intra- and extracellular electrolyte concentrations were measured by electron microprobe analysis, tissue contents of methylamines (glycerophosphorylcholine, betaine), polyols (myo-inositol, sorbitol), and several amino acids in different kidney zones by high-performance liquid chromatography. Administering DDAVP continuously after diuresis increased Uosm from (means +/- SE) 348 +/- 8 to 1,265 +/- 127 after 1 day and 2,485 +/- 186 mosmol/kgH2O after 3 days. The sum of all osmolytes at the papillary tip rose from 309.2 +/- 28.9 to 690.9 +/- 105.8 and 1,282.8 +/- 21.0 mmol/kg protein after days 1 and 3, respectively. Although interstitial tonicity (sum of Na, Cl, and K concentrations) was increased by 116 and 223% after 1 and 3 days DDAVP, intracellular tonicity was similar in chronic diuresis and following 1 or 3 days DDAVP. Coadministration of DDAVP with betaine, myo-inositol, and choline ("osmolyte treatment") did not accelerate the restoration of Uosm but caused significantly higher contents of osmolytes (except myo-inositol) in inner medulla and/or papilla after 3 days. In a minority of animals, restoration of Uosm and reaccumulation of medullary osmolytes were impeded in both DDAVP- and DDAVP/osmolyte-treated rats. These data indicate that, after chronic diuresis, accumulation of organic osmolytes and restoration of Uosm proceed in parallel. Capacity for transport and/or synthesis of organic osmolytes, rather than their availability, appear to limit reaccumulation on the first day of recovery. By the third day, delivery of some osmolytes or their precursors may limit the restoration of medullary osmolyte content. The failure of some rats to attain sufficient concentrating ability within this time period may be related to deficient reaccumulation of medullary osmolytes. PMID- 7485533 TI - Prevention of experimental cyclosporin-induced interstitial fibrosis by losartan and enalapril. AB - The pathogenesis of renal scarring in chronic cyclosporin nephropathy is unknown. In this study, we evaluated the effects of renin-angiotensin system blockade by enalapril and losartan in a salt-dependent model of cyclosporin-associated chronic tubulointerstitial fibrosis (TIF). Rats kept on normal or low-salt diet were given cyclosporin, cyclosporin+enalapril, cyclosporin+losartan, cyclosporin+enalapril#losartan, or vehicle for 14 and 28 days. Cyclosporin reduced glomerular filtration rate (GFR) in rats fed either diet, but only salt depleted animals developed significant TIF. Cyclosporin also impaired renal concentrating ability and caused tubular enzymuria. Renin-angiotensin system blockade decreased blood pressure (BP) and promoted afferent arteriolar vasodilatation. Losartan reduced plasma renin activity and prevented cyclosporin induced increment of cortical alpha 1(I) procollagen mRNA. Renin-angiotensin blockade did not improve GFR and tubular function; however, it strikingly prevented TIF development, even in presence of very low BP. Rats treated with cyclosporin, hydralazine, and furosemide achieved BP values similar to losartan or enalapril groups, but there was no protection against interstitial fibrosis development. These results suggest that cyclosporin-related chronic interstitial injury is mediated by angiotensin II and that the mechanisms promoting the interstitial scarring can be dissociated from glomerular and tubular dysfunction in cyclosporin nephropathy. PMID- 7485535 TI - Effects of chronic Cl depletion alkalosis on proximal tubule transport and renal production of ammonium. AB - The role of renal ammonium excretion in the maintenance of chronic metabolic alkalosis is poorly defined, particularly under conditions in which the alkalosis is associated with secondary potassium depletion. Therefore, free-flow micropuncture experiments were performed to examine the effects of chronic chloride depletion metabolic alkalosis (CDAlk) on renal ammonium production, urinary ammonium excretion, and proximal convoluted tubule (PCT) ammonium transport in the rat in vivo. CDAlk was generated by peritoneal dialysis against NaHCO3 and maintained for 6-7 days by dietary Cl- restriction. Pair-fed controls were dialyzed against NaCl. Rats with CDAlk had elevated plasma HCO3- concentration, hypokalemia, and hypochloremia. HCO3- excretion was negligible in both control and CDAlk rats. Glomerular filtration rate and urine pH did not differ. CDAlk reduced urinary ammonium excretion by 35% but had no significant effect on whole kidney ammonium production. Net secretion of ammonium by the PCT was decreased by 70% and absolute delivery of ammonium out of the PCT was decreased by 55% in the CDAlk rats. The decrease in PCT ammonium secretion was the combined result of a decrease in net ammonium secretion along the early PCT and an increase in net ammonium absorption along the late PCT.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7485536 TI - Autoregulation of zonal glomerular filtration rate and renal blood flow in spontaneously hypertensive rats. AB - Autoregulation of total and zonal glomerular filtration rate (GFR) in outer, middle and inner cortex was estimated in spontaneously hypertensive rats (SHR), from the tubular uptake of 125I-labeled aprotinin (125I-Ap), injected at control renal arterial pressure (RAP), and 131I-Ap, injected at reduced RAP in left kidney. Normotensive Wistar-Kyoto (WKY) rats were used as controls. Renal blood flow (RBF) autoregulation was reset to higher pressure levels in SHR. When RAP was lowered close to the lower pressure limit of RBF autoregulation, total GFR was reduced to 89.5 +/- 3.1 and 88.1 +/- 3.3% of control in 10- and 40-wk WKY and to 87.7 +/- 2.3 and 88.0 +/- 2.2% in 10- and 40-wk SHR. In WKY, the fall of GFR in the three cortical layers was not different during RAP reduction. In 10- and 40-wk-old SHR, however, GFR fell significantly less in inner than in middle and outer cortex (P < 0.05). We conclude that autoregulation of GFR is most efficient in the inner cortex of SHR. In all animals, GFR was less well autoregulated than RBF. PMID- 7485537 TI - Expression of endothelins 1 and 3 in the rat kidney. AB - Endothelins (ETs) 1 and 3 are expressed in the rat kidney, but the factors that regulate this expression remain unknown. To try to understand what these might be, we have measured the renal levels of ET-1 and ET-3 mRNAs by the ribonuclease protection-assay technique after a number of clearly defined renal/hemodynamic insults. 1) Six hours after the induction of hemorrhagic anemia and hypotension, there was a threefold increase in ET-1 mRNA and a simultaneous threefold decrease in ET-3 mRNA. This indicates that, in this situation, these two ET isoforms are differentially controlled and emphasizes the need for assay techniques capable of distinguishing between them. 2) One day after application of a 0.2-mm clip to the left renal artery, there was a > 2.5-fold induction of ET-1 mRNA in that kidney, which persisted for 10 days. A smaller rise in ET-1 mRNA was seen in the contralateral organ. After 2 days, ET-3 mRNA levels were reduced by approximately 50% in the clipped organ. Both ramipril (an angiotensin-converting enzyme inhibitor, 7.5 mg/kg daily) and bosentan (a nonselective ET receptor antagonist, 100 mg/kg daily) substantially reduced the elevation in ET-1 mRNA seen in the clipped kidney after 2 days, suggesting that the generation of angiotensin II and the action of ET itself are involved in the mechanism by which clipping stimulates ET-1 expression. By contrast, ramipril, but not bosentan, prevented the reduction in ET-3 mRNA levels. 3) Renal denervation, dietary salt restriction, or diuretic treatment (furosemide) did not alter renal expression of ET-1 or ET-3.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7485538 TI - Increased sensitivity of the renal vasculature to adenosine in streptozotocin induced diabetes mellitus rats. AB - Adenosine (ADO) has been implicated as a pathophysiological factor in contrast media (CM)-induced acute renal failure, which has been encountered more often in patients with diabetes and impaired renal function. Therefore, we studied the renal vascular response to exogenous and endogenous ADO in streptozotocin-induced diabetic rats. We found that exogenous ADO (0.01-100 nmol), injected into the abdominal aorta, decreased renal blood flow (RBF) in a dose-dependent manner. The dose-response curve was shifted to the left by factor 30 in diabetic, compared with nondiabetic rats rats. Renal vascular response to endogenous ADO, assessed by postocclusive reduction of RBF after a 30-s renal artery occlusion, was significantly enhanced (P < 0.001) in diabetic rats (75.6 +/- 3.9%) compared with nondiabetic rats (36.5 +/- 2%). ADO A1-receptor blockade with 8-cyclopentyl-1,3 dipropylxanthine attenuated exogenous and endogenous ADO-induced renal vasoconstriction in both groups. We conclude that the ADO A1-receptor signal transduction chain is altered in diabetic animals and that the enhanced vasoconstrictive action of ADO could be involved in the kidney pathophysiology of diabetes mellitus. PMID- 7485539 TI - Heterogeneity of NH+4 transport in mouse inner medullary collecting duct cells. AB - Previous studies from our laboratory have demonstrated that NH+4 substitutes for K+ on the Na(+)-K(+)-adenosinetriphosphatase (Na(+)-K(+)-ATPase) in rat terminal inner medullary collecting duct cells (tIMCD). To examine other NH+4 transport pathways, a transgenic mouse cell line, mIMCD-3, was employed. 86Rb+ was used as a K+ congener to explore NH+4/Rb+ (K+) competition on the extracellular K+ binding site of the Na(+)-K(+)-2Cl- cotransporter and the Na(+)-K(+)-ATPase. Addition of K+ or NH+4 reduced both bumetanide- and ouabain-sensitive Rb+ uptake. This reduction in Rb+ uptake with NH+4 addition was not due to intracellular pH mediated changes in transporter activity. K+ and NH+4 are competitive inhibitors on both transporters. On the Na(+)-K(+)-2Cl- cotransporter, the Michaelis constant (Km) for K+ was 4.6 +/- 0.5 mM with an inhibitory constant (Ki) for NH+4 of 2.8 mM. In contrast, on the Na(+)-K(+)-ATPase, the apparent affinity for K+ was greater than for NH+4. To test Na(+)-K(+)-2Cl- cotransport-mediated NH+4 flux, bumetanide-sensitive NH+4/Rb+ exchange was measured. Bumetanide-sensitive Rb+ efflux was greater with extracellular K+ or NH+4 present relative to efflux with extracellular N-methyl-D-glucamine. This demonstrates both K+/Rb+ and NH+4/Rb+ countertransport by the Na(+)-K(+)-2Cl- cotransporter. In conclusion, NH+4 is transported in a bumetanide-sensitive Na(+)-NH+4-Cl- mode, and both NH+4 and Rb+ (K+) are competitive inhibitors for the extracellular K+ binding site. However, the kinetics of Na(+)-K(+)-2Cl(-)-mediated NH+4 transport differ from other K+ transport-mediated NH+4 pathways, such as the Na(+)-K(+)-ATPase. PMID- 7485541 TI - Expression of colonic H-K-ATPase mRNA in cortical collecting duct: regulation by acid/base balance. AB - In addition to the gastric isoform of H-K-ATPase, the colonic isoform is also expressed in the kidney, but its intrarenal localization and exact function are not known. The goal of this study was to determine whether the colonic H-K-ATPase is expressed in the rabbit cortical collecting duct (CCD) and whether it is regulated by changes in acid/base balance. With quantitative reverse transcriptase-polymerase chain reaction (RT-PCR) with RNA isolated from immunodissected rabbit CCD cells and degenerate oligonucleotide primers, a PCR product of the predicted size (approximately 430 bp) was amplified. The amplified DNA was further characterized by nested PCR and sequencing. Direct sequencing of the 434-bp PCR product revealed 83% identity at the nucleotide level and an 80.4% identity at the deduced amino acid level to the rat colonic H-K-ATPase. With the same primers and cDNA originating from rabbit distal colon, a DNA fragment with a size and nucleotide sequence identical to that originating from CCD cells was amplified. Furthermore, using PCR screening, we isolated and sequenced a 1.5-kb cDNA clone from a rabbit CCD library. The predicted amino acid sequence of the protein encoded by this cDNA is 85 and 82% identical to the corresponding regions of the guinea pig and rat colonic H-K-ATPase, respectively, and 70% identical to the H-K-ATPase recently cloned from Bufo marinus, whereas it shows only 45 and 42% homology to the rat Na-K-ATPase alpha 1-subunit and the rat gastric H-K ATPase, respectively.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7485540 TI - NH3 permeability of principal cells and intercalated cells measured by confocal fluorescence imaging. AB - The cortical collecting duct (CCD) is an important site for NH3 secretion in mammalian nephron. However, given the cellular heterogeneity of this epithelium, the transcellular sites for NH3 secretion are unknown. In the present study, a dual-excitation confocal microscope was designed and optimized to have sufficient temporal resolution to measure the permeability of ammonia (PNH3) across the basolateral and apical membrane of principal cells (PCs) and intercalated cells (ICs) in perfused rabbit CCDs. The rate of cellular NH3 influx was calculated from the time course of increase in intracellular pH (pHi), measured with 2',7' bis(carboxyethyl)-5(6)-carboxyfluorescein after 20 mM NH4Cl was added to the bath or luminal perfusate. The time course of increase in pHi was calculated from 488/442 image pairs stored at a rate of 4 Hz. The apparent basolateral and apical PNH3 values of PCs were 36 +/- 5 and 113 +/- 11 microns/s, respectively. The values were 5.0 +/- 0.7 and 34 +/- 3 microns/s after membrane folding correction. The apparent basolateral and apical PNH3 values of ICs were 38 +/- 6 and 132 +/- 15 microns/s. Corrected for membrane folding, the values were 9.0 +/- 1.0 and 47 +/- 5 microns/s, respectively. The results demonstrate that the apical surface was more permeable than the basolateral surface in both cell types. In addition, ICs were more permeable to NH3 than PCs across both membranes. The transcellular PNH3 of PCs and ICs were 27.3 and 29.5 microns/s, respectively.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7485542 TI - Expression of calcineurin activity and alpha-subunit isoforms in specific segments of the rat nephron. AB - Calcineurin activity and alpha-subunit expression were studied in microdissected proximal tubules (S2), medullary thick ascending limbs (MTAL), cortical collecting ducts (CCD), connecting tubules (CNT), and outer medullary collecting ducts (OMCD). We have shown that cyclosporin A (CsA) and FK-506 inhibit sodium potassium-adenosinetriphosphatase (Na-K-ATPase) activity in CCD, OMCD, and MTAL but did not uncover the mechanism for resistance of proximal tubule segments to these drugs. Because cells expressing high calcineurin activity are relatively resistant to the biological effects of CsA and FK-506, we hypothesized that the resistance of proximal tubules may be linked to increased calcineurin expression. Consequently, we measured calcineurin activity in microdissected tubules using a calcineurin-specific substrate. Calcineurin activity in S2 proximal tubule segments was 10-fold higher than in CCD, CNT, OMCD, or MTAL. FK-506 (6.0 ng/ml) inhibited calcineurin activity in CCD, CNT, and MTAL but not S2; 250 ng/ml FK-506 inhibited S2 calcineurin activity by 50%. Likewise, high concentrations of CsA (25 micrograms/ml) and FK-506 (250 ng/ml) inhibited S2 Na-K-ATPase activity by 77 and 73%, respectively. To investigate whether the resistance of S2 segments might be due to differential expression of calcineurin alpha-subunit isoforms, we determined the isoform expression by Western blot analysis using isoform-specific antibodies against the alpha 1-, alpha 2-, and alpha 3-isoforms. We found that alpha 1 expression in S2 was significantly greater than in the CCD and MTAL, whereas alpha 2 expression in the S2 was significantly less than in CCD and MTAL. No alpha 3 was detected in any nephron segment tested.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7485543 TI - DARPP-32 promoter directs transgene expression to renal thick ascending limb of loop of Henle. AB - DARPP-32, a dopamine- and adenosine 3',5'-cyclic monophosphate (cAMP)-regulated inhibitor of protein phosphatase-1, is highly colocalized with neuronal and nonneuronal D1-type receptors. DARPP-32 concentration is enriched in the renal outer medulla and in the medium-size spiny neurons of the brain. In the ascending limb of the loop of Henle, DARPP-32 is phosphorylated following stimulation by dopamine and other first messengers, and in this form inhibits the activity of the Na(+)-K(+)-adenosinetriphosphatase pump. For functional analysis of the DARPP 32 promoter in the kidney, we characterized the murine gene. There are two groups of transcription start sites utilized in the brain, but the proximal set appears to be preferentially used in the kidney. In four of four lines of mice carrying a DARPP-32/lacZ transgene with 2.1 kb of 5'-flanking DNA, adult kidney lacZ transgene expression mimicked that of endogenous DARPP-32. There was no ectopic expression in peripheral organs. We conclude that the sequences necessary for direction of DARPP-32 expression to the medullary thick ascending limb are contained within this 2.1-kb fragment. PMID- 7485544 TI - Noninvasive measurement of intrarenal blood flow distribution: kinetic model of renal 123I-hippuran handling. AB - A new technique for noninvasive measurement of intrarenal blood flow distribution over cortex and medulla is proposed. The technique involves analysis of 123I labeled hippuran renography, according to a kinetic model that describes the flow of 123I-hippuran from the heart (input) through the renal cortex and medulla to the bladder (output). The method is validated and compared with the standard microsphere injection technique in anesthetized dogs. Changes in intrarenal blood flow distribution were induced by infusion of placebo (n = 6), angiotensin I (n = 5), or atrial natriuretic factor (n = 5). Baseline percentage medullary blood flow in the left kidney was 12 +/- 1% of total renal blood flow measured with microspheres and 15 +/- 1% with renography. During infusion of the placebo, medullary blood flow decreased slightly compared with baseline, as measured with both methods, by 2 +/- 6 (microspheres) and 1 +/- 8% (renography). Infusion of angiotensin I caused a marked fall in medullary blood flow by 42 +/- 11 (microspheres) and 57 +/- 8% (renography). In contrast, infusion of atrial natriuretic factor caused a small rise in medullary blood flow as measured with both methods (9 +/- 3 and 12 +/- 11%, respectively). The absolute and percent changes in medullary blood flow measured with renography correlated with those measured with microspheres (left kidney: r = 0.67, P = 0.005; r = 0.71, P = 0.003, respectively; right kidney: r = 0.62, P = 0.01; r = 0.68, P = 0.004, respectively). We conclude that the proposed kinetic model of renal 123I-hippuran handling can be used to measure changes in intrarenal blood flow distribution and, because of its noninvasive character, may be of use in clinical studies. PMID- 7485545 TI - Analysis of interaction between TGF and the myogenic response in renal blood flow autoregulation. AB - The present study investigates the interaction between the tubuloglomerular feedback (TGF) response and the myogenic mechanism by use of a mathematical model. The two control mechanisms are implemented in a spatially distributed model of the rat renal juxtamedullary afferent arteriole. The model of the afferent arteriole is based on in vivo measurements of the stress-strain relation in muscle strips. Analysis of experimental data shows that the myogenic response can be modeled by a linear relation between the transmural pressure and the level of activation of the vascular smooth muscle cells. The contribution of TGF to smooth muscle activity is assumed to be a linear function of the glomerular capillary pressure. The results show that the myogenic response plays an important role in renal blood flow autoregulation. Without a myogenic response, mechanisms such as TGF that are localized in the distal segments of the microvasculature would not be able to achieve autoregulation because of passive, pressure-mediated effects in the upstream vascular segments. In addition, it is shown that a strong myogenic response may lead to both propagation and enhancement of vascular effects mediated through mechanisms located in the distal part of the afferent arteriole. An ascending myogenic response could enhance the regulatory efficiency of the TGF mechanism by increasing the open-loop gain of the system. However, such a synergistic interaction will only be observed when the two mechanisms operate on more or less separate segments of the afferent arteriole. In the case where they operate on common segments of the arteriole, the outcome of the interaction may well be antagonistic. PMID- 7485546 TI - Tyrosine phosphorylation is a novel pathway for regulation of chloride secretion in shark rectal gland. AB - We used the specific tyrosine kinase inhibitor genistein to define the involvement of tyrosine phosphorylation in the regulation of chloride transport in the rectal gland of the dogfish shark, a model for chloride secretion via a cystic fibrosis transmembrane conductance regulator (CFTR)-like channel. In the perfused gland, genistein (100 microM) promptly increased chloride secretion from basal values of 159 +/- 36 to 966 +/- 49 mueq.h-1.g-1 (P < 0.0001). Bumentanide fully reversed genistein-induced secretion. In primary culture monolayers of rectal gland tubular cells, genistein, but not the inactive 7-glucoside form, genistin, increased short-circuit current in a dose-dependent manner, from basal values of 2.7 +/- 4.3 to 104 +/- 10 microA/cm2 (P < 0.0001). Apically applied genistein induced significantly greater chloride secretion than basolateral addition. Genistein did not increase the adenosine 3',5'-cyclic monophosphate (cAMP) content of either perfused glands or cultured monolayers. Using an anti phosphotyrosine antibody, we observed phosphorylation of multiple proteins. Four peptides, with molecular masses of 250, 210, 55, and 53 kDa, responded to genistein treatment with a decrease in tyrosine phosphorylation. These data demonstrate the following: 1) genistein induces bumetanide-sensitive chloride secretion in both perfused rectal glands and cultured tubular cells; 2) these effects are not accompanied by an elevation of tissue cAMP, indicating that genistein-induced secretion is not mediated by the cAMP-protein kinase A pathway; and 3) genistein-sensitive peptides are present in the rectal gland cell and are candidates for involvement in the regulation of chloride secretion.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7485547 TI - Coronary flow stimulates auricular-ventricular transmission in the isolated perfused guinea pig heart. AB - In the heart in situ coronary flow stimulates oxygen consumption, glycolytic flux, myocardial contractility, and the release of bioactive substances. Studies have indicated that the coronary flow-enhanced contraction is similar to a hormonelike effect because the enhanced contraction results from an elevation in intracellular free calcium. In fact, if extracellular calcium is raised sufficiently, the contraction amplitude rises and remains constant and independent of coronary flow. We hypothesized that coronary flow could also stimulate other calcium-dependent cardiac functions such as auricular-ventricular (A-V) transmission. This hypothesis was tested in isolated guinea pig hearts perfused at constant flow. Our results show that increases in coronary flow (6-25 ml/min range) decrease the A-V delay solely as a result of reduced propagation time in the A-V node and not in atrial or ventricular propagation. When coronary vascular resistance was altered by dilation (nitroglycerin, bradykinin, nitroprusside, and adenosine) or by constriction (angiotensin II), this dromotropic effect of flow remained the same despite wide changes in perfusing pressure. Also, this dromotropic effect of flow was not altered by energy altering substrates in the perfusate or by perfusion of adenosine receptor blockers. Furthermore, the effectiveness of flow as a dromotropic stimulus varied inversely with changes in calcium entry caused either by elevation or reduction of extracellular calcium. In addition, enhanced viscosity of the perfusing medium amplifies the positive dromotropic effect of flow. These results suggest that coronary flow is a stimulus that exerts a positive dromotropic effect mediated by shear stress. PMID- 7485548 TI - Role of angiotensin-converting enzyme and angiotensin II in development of hypoxic pulmonary hypertension. AB - Although angiotensin converting enzyme (ACE) inhibitors are known to attenuate the development of hypoxic pulmonary hypertension in rats, the precise mechanism of this protective effect remains unknown. Thus we utilized specific angiotensin II (ANG II)-receptor antagonists to investigate whether ANG II is involved directly in the hemodynamic and structural changes of pulmonary hypertension, and we tested whether the protective effects of ACE inhibition can be attributed partly to potentiation of bradykinin. During 14 days of hypobaric hypoxia, rats received, via intraperitoneal osmotic minipumps, either 1) the ACE inhibitor captopril, 2) captopril plus the bradykinin B2-receptor antagonist CP-0597, 3) the ANG II type 1 receptor antagonist losartan, 4) the ANG II type 2 receptor antagonist PD-123319, or 5) saline. At 14 days, mean pulmonary arterial pressure (MPAP) was reduced (P < 0.05) in hypoxic rats treated with captopril (26.6 +/- 0.8 mmHg) or losartan (24.4 +/- 1.0 mmHg) compared with saline (32.0 +/- 1.4 mmHg) but was unaffected by PD-123319 (29.5 +/- 1.7 mmHg). Right ventricular hypertrophy was reduced in hypoxic rats treated with captopril or losartan compared with saline-treated rats. Morphometry showed less medial thickening and peripheral muscularization of small pulmonary arteries in hypoxic animals treated with captopril or losartan. Coadministration of CP-0597 did not reverse the protective effects of captopril on pulmonary vascular remodeling. These results suggest a novel role for endogenous ANG II, acting through the type 1 receptor, in the vascular remodeling associated with hypoxic pulmonary hypertension. The beneficial effects of ACE inhibition in this model can be attributed to reduced ANG II production rather than potentiation of bradykinin. PMID- 7485549 TI - NBT reactivity correlates to the distribution of PMNs between rat pulmonary and systemic circulation. AB - The rat systemic and pulmonary vascular transit times of radiolabeled polymorphonuclear granulocytes (experimental population referred to as "Polys") and mononuclear leukocytes (experimental population referred to as "Monos") (111In) in relation to those of erythrocytes (51Cr) were measured under physiological conditions. Results were also expressed as "circulating" pool and "marginated" pool of the two leukocyte fractions, and it was investigated whether the degree of spontaneous granulocyte activation, measured with the nitro blue tetrazolium (NBT) reduction test, was of importance for the retention of Polys in the lungs. The measured mean vascular transit was similar for Monos and Polys and several times slower than that of the erythrocytes in the pulmonary (17.4 times) and the systemic (4.1 times) vascular beds. The percentage of NBT-positive cells was < 10% in most animals, indicating a low level of spontaneous granulocyte activation. Increasing levels of NBT-positive cells correlated with a redistribution of granulocytes from the systemic marginated cell pool to the pulmonary marginated cell pool. Microscopic evaluation of sections from embedded lung tissue biopsies, obtained after intravital staining of the leukocytes, confirmed the isotope data on pulmonary transit. PMID- 7485550 TI - Changes in contractility and afterload have only slight effects on subendocardial systolic flow impediment. AB - To test the hypothesis that contractility did not greatly influence systolic impediment to subendocardial flow we perfused the coronary artery at 70 mmHg and altered aortic pressure in 7 dogs and contractility in another 7. We measured myocardial systolic flow impediment (SFI) by comparing regional flows while beating and during asystole. Cardiac contraction impeded 29% of subendocardial asystolic flow, which was not affected by either intervention. In subepicardium, contraction increased flow by 25%, but dobutamine impeded systolic flow. Subepicardial SFI was only 16% of subendocardial SFI. Dobutamine slightly decreased estimated percent systolic myocardial blood flow (%SMBF) in subendocardium (+/- 12%) but decreased subepicardial %SMBF (45.5 to 17.4%). Phasic coronary flow pulsatility increased more with dobutamine than increased afterload, and pulsatility and SFI correlated only in subepicardium. Systolic-to total coronary flow ratio and %SMBF did not correlate closely in subendocardium. SFI was most prominent in the subendocardium, whereas subepicardial SFI mainly determined epicardial coronary flow pulsatility. We conclude that the effects of contractility changes differ when evaluating regional SFI vs. phasic flow pulsatility. PMID- 7485551 TI - Age-related changes in perimicrovascular protein distribution. AB - The diffusion hypothesis for physiological aging proposes that an increase in interstitial matrix fiber-to-gel ratio causes a decrease in nutrient diffusion to the cells. This hypothesis predicts a decrease in interstitial matrix protein with age. The objective was to test this hypothesis by determining age-related changes in plasma protein distribution in perimicrovascular and distal regions of rat mesentery interstitial matrix. Rats that were 77, 140, 210, 315, 455, and 630 days old were anesthetized with pentobarbital sodium, and a mesenteric loop was exteriorized. Intravital video microspectrophotometry was performed using wavelengths of 280, 320, and 700 nm. Perimicrovascular protein concentrations from the protein absorbance images were used to obtain the histogram, mean, and skewness of the proximal and distal protein concentration distributions. An exponential gradient model was also used to obtain the proximal and distal protein concentrations and gradient decay constants. Proximal protein concentration increased from 77- to 140-day-old rat and then decreased gradually through 210-, 315-, 455-, and 630-day-old rats. Distal concentration decreased gradually from 140- to 630-day-old rats. There was an increase in positive skewness of the proximal protein distributions from 140- through 630-day-old rats. We found an age-related decrease in perimicrovascular protein and propose that this is due to a decrease in protein permeability with age. The results support the diffusion theory of aging. PMID- 7485552 TI - Captopril-induced hypotension is inhibited by the bradykinin blocker HOE-140 in Na(+)-depleted marmosets. AB - Inhibition of angiotensin-converting enzyme (ACE) inhibits formation of angiotensin II and, by inhibition of kinin metabolism, may also increase vascular bradykinin. The present experiments were done in sodium-depleted, conscious, unrestrained marmosets (n = 5-11) to examine the contribution of bradykinin to ACE inhibitor-induced hypotension. Aortic blood pressure and heart rate (HR) were monitored via telemetry. After sodium depletion (low-sodium diet and furosemide), captopril (1 mg/kg po) caused a significant (P < 0.05) decrease in mean arterial blood pressure (MABP) (-34 +/- 3 mmHg, maximally, from 79 +/- 2 mmHg) but no change in HR compared with vehicle treatment. The bradykinin receptor antagonist HOE-140 (1 mg/kg sc) significantly inhibited the hypotensive response to captopril and caused marked tachycardia (+133 +/- 14 beats/min from 214 +/- 8 beats/min). HOE-140 (1 mg/kg sc) followed by vehicle administration had no effect on MABP but increased HR similarly. The hypotensive response to captopril was inhibited by HOE-140 regardless of the order of administration or the route of captopril administration (by mouth vs. subcutaneously). The hypotensive response to a renin inhibitor, A-72517 (3 mg/kg sc), was not inhibited by prior HOE-140 administration despite a similar HOE-140-induced tachycardia. These data suggest that the hypotensive effect of captopril in sodium-depleted, conscious marmosets is dependent on functional bradykinin B2 receptors. Also, blockade of B2 receptors uncovers marked tachycardia in this model, suggesting a tonic effect of bradykinin on control of HR in marmosets. PMID- 7485553 TI - Direct detection and quantification of singlet oxygen during ischemia and reperfusion in rat hearts. AB - To detect singlet oxygen (1O2) in postischemic reperfused hearts, 5,8 endoperoxide, an oxidation product of beta-carotene, was used as a marker for 1O2 generation and was quantified using high-performance liquid chromatography (HPLC). Isolated rat hearts were subjected to ischemia for 5, 10, 20, 30, and 60 min followed by 10 min of reperfusion with buffer containing 25 microM beta carotene. The coronary effluent was collected, extracted, and injected into the HPLC unit. The production of 5,8-endoperoxide was maximum during the first 2 min of reperfusion. Maximal accumulated amount of 1O2 was observed in hearts subjected to 60-min ischemia (36.2 +/- 1.7 nmol.10 min-1.g-1) as compared with 10 min ischemia (6.2 +/- 1.0 nmol.10 min-1.g-1). There was a good correlation between the amount of 1O2 production and cardiac function. Treatment with 25 mM histidine significantly decreased 5,8-endoperoxide from 7.02 +/- 0.47 to 0.98 +/- 0.11 nmol.min-1.g-1 (P < 0.01) and improved cardiac function in the group with 60 min ischemia. This study demonstrates that 1) the present method is useful and reliable for the measurement of 1O2 in the heart, 2) 1O2 production during reperfusion is dependent on the duration of initial ischemia, and 3) 1O2 is one of the major factors in postischemic reperfusion injury. PMID- 7485554 TI - Downward shift of coronary pressure-flow relationship following a brief period of ischemia in dogs. AB - This study was undertaken to test whether a brief period of ischemia affects the coronary pressure-flow relationship during reduction of coronary perfusion pressure (CPP). The left anterior descending coronary artery was cannulated and perfused with blood from the left carotid artery in 40 open-chest dogs. Coronary blood flow (CBF) was measured during intracoronary administrations of papaverine and adenosine. The coronary pressure-flow relationship was assessed during transient reduction of CPP from 100 to 30 mmHg. Coronary hyperemic flow due to adenosine and papaverine was attenuated 30 min after transient 10- and 15-min periods of ischemia. In the group of transient 10-min ischemia, both fractional shortening (FS) and CBF returned to the preischemic values at 30 and 60 min of reperfusion; however, marked decreases in CBF (35 +/- 5 vs. 56 +/- 4 ml.100 g 1.min-1 at CPP = 60 mmHg, P < 0.01) during graded reductions in CPP were observed. The endomyocardial blood flow was reduced relative to the control condition. Furthermore, both FS (6 +/- 1 vs. 14 +/- 1% at CPP = 60 mmHg, P < 0.01) and lactate extraction ratio (-41 +/- 15 vs. 1 +/- 6% at CPP = 60 mmHg, P < 0.05) were decreased. The downward shift of the CPP-CBF relationship and the deterioration of myocardial contractile and metabolic function during reduction of CPP were restored 60 min after the onset of reperfusion.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7485556 TI - Mechanisms of conduction time hysteresis in rabbit atrioventricular node. AB - The functional origin of atrioventricular nodal hysteresis was studied in isolated rabbit heart preparations. This hysteresis is characterized by asymmetric changes in nodal conduction time (NCT) occurring for symmetric changes in cycle length. The respective contribution of the nodal properties of recovery, facilitation, and fatigue to the beat-to-beat changes in NCT observed during paired symmetric ramps of decreasing and increasing cycle length was determined with specifically design stimulation protocols. Nodal hysteresis was found to be entirely accounted for by variations in the contribution of nodal recovery and fatigue properties observed at corresponding cycle lengths. The study establishes how this contribution varies on a beat-to-beat basis as a result of cycle length history. This holds true for the numerous changes in hysteresis observed in response to changes in the sequence and slope of the ramps. Facilitation clearly affected NCT during these responses but did not contribute to the hysteresis. Moreover, the study demonstrates that there is no inherent change in the characteristics of nodal function with the direction of the ramp that could account for the hysteresis. Thus nodal hysteresis arises from nodal functional properties of recovery and fatigue but does not constitute a distinct independent intrinsic property of the node. PMID- 7485555 TI - Ca(2+)-dependent K+ current in arterial smooth muscle cells from aldosterone-salt hypertensive rats. AB - Aorta from aldosterone-salt hypertensive rats (AHR) demonstrates an increased basal 42K efflux. We investigated the cellular mechanisms of this alteration by measuring 42K efflux from aortic segments as well as myoplasmic Ca2+ concentration ([Ca2+]m) and K+ current in aortic smooth muscle cells from AHR and normotensive control-salt rats (CSR). Both diltiazem and nisoldipine attenuated but did not normalize the increase in basal 42K efflux in AHR. The resting [Ca2+]m was elevated in cells from AHR (148 +/- 15 vs. 91 +/- 12 nM for CSR, P < 0.05). The rate of Mn2+ quenching under basal conditions was also increased in cells from AHR, and the increase was abolished by Cd2+. However, the resting membrane potential did not differ between CSR and AHR (-49 +/- 5 vs. -50 +/- 4 mV). The steady-state, whole cell K+ current was also increased in cells from AHR. This increase was abolished by charybdotoxin, tetraethylammonium, La3+, and by clamping [Ca2+]m at zero or 100 nM with ethylene glycol-bis(beta-aminoethyl ether)-N,N,N',N'-tetraacetic acid. The single-channel conductance of the large conductance Ca(2+)-activated, voltage-dependent K+ (KCa) channels was not altered in AHR. Further, 33% of cells from AHR vs. 1% from CSR showed spontaneous transient outward K+ currents, which reflect activation of KCa channels by Ca2+ released from caffeine-sensitive stores. While the acute caffeine-induced [Ca2+]m response was similar between CSR and AHR, the outward current and 42K efflux responses to caffeine were greater in AHR. After continued exposure to caffeine, the basal 42K efflux was attenuated more in AHR than in CSR. Charybdotoxin resulted in a greater depolarization in AHR cells than in CSR cells (9.8 +/- 2.2 vs. 3.5 +/- 1.6 mV, P < 0.05). These results indicate that the increases in both 42K efflux and K+ current reflect an increased activity of KCa channels that is associated with an increased Ca2+ influx and resting [Ca2+]m and altered Ca2+ handling by the sarcoplasmic reticulum in aortic smooth muscle cells from AHR. PMID- 7485557 TI - Activation of angiotensin-converting enzyme expression in infarct zone following myocardial infarction. AB - In the present study we quantified angiotensin-converting enzyme (ACE) mRNA and localized ACE mRNA and protein in the infarcted rat heart. Wistar rats underwent ligation of the left descending coronary artery, resulting in myocardial infarction (MI) or a sham operation. At different times (1-90 days) after surgery (n = 3 each), the heart was removed and divided into the right ventricle (RV), septum (Se) and left ventricle (LV). ACE mRNA was quantified by competitive reverse transcriptase-polymerase chain reaction (RT-PCR). At 4 and 7 days after MI, we found a 2.8-fold increase of ACE mRNA (n = 3; P < or = 0.05) in the infarcted LV compared with the LV of the sham group. No increases of ACE mRNA were found in the noninfarcted hypertrophied compartments. ACE activity increased 2.6- and 3.6-fold in the infarcted LV at 7 and 90 days after MI, respectively. In situ hybridization and immunohistochemistry showed increased ACE mRNA and protein density in the border zone of the infarcted area, predominantly in the endothelial cells lining capillaries. In the noninfarcted myocardium ACE mRNA and protein were confined to endothelial cells of the larger vessels. From these data we conclude that the intracardiac RAS is involved in the healing of the scar after MI in the rat, possibly giving rise to neovascularization. Furthermore, the data suggest that the intracardiac ACE is not necessarily associated with hypertrophy in the rat heart after MI. PMID- 7485558 TI - Thoracic duct lymph flow responses to hemorrhage in the ovine fetus. AB - Blood volume returns toward normal after hemorrhage much more rapidly in the fetus than in the adult due to a rapid entry of fluid and plasma proteins into the fetal circulation. One potential source of fetal fluids and plasma proteins is the lymphatic system, since basal lymph flow rate and interstitial protein concentration are high in the fetus. Furthermore, studies in adults suggest that lymph flow rate may increase following hemorrhage. To test the hypothesis that hemorrhage induces an increase in lymph flow in the fetus, 15 late-gestation ovine fetuses underwent left thoracic duct catheterization with low-resistance catheters and were studied 5 or more days after surgery at 134 +/- 1 (SE) days gestation. The protocol included three successive 30-min periods: control, hemorrhage, and recovery. During the first 5 min of the hemorrhagic period, 61 +/ 4 ml of fetal blood were removed. The blood was reinfused over the first 5 min of the recovery period. After the hemorrhage, fetal arterial pressure, venous pressure, and heart rate decreased (analysis of variance, P < 0.001). These variables significantly increased above basal levels following blood reinfusion. Fetal hematocrit (P < 0.001) and plasma protein concentration (P < 0.05) also decreased after the hemorrhage and returned to control values after the reinfusion. Fetal lymph flow rate was 0.55 +/- 0.06 (SE) ml/min before the hemorrhage and decreased by a maximum of 30.3 +/- 6.3% (P < 0.001) at 8 min after the end of the hemorrhage. Lymph flow rate was reduced by an average of 19.1 +/- 6.6% during the hemorrhagic period and returned to prehemorrhage levels following blood reinfusion.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7485559 TI - Recognition of consensus CHO structure in ligands for selectins by novel antibody against sialyl Lewis X. AB - The selectins (L, E, and P) play an important role in the earliest events of the inflammatory response, leading to the "rolling" phenomenon. All selectins react with sialyl Lewis X (SLex) in vitro, possibly suggesting that their ligands have a consensus structure. 2H5 is a monoclonal antibody against SLex that blocks L selectin-mediated adhesion. 2H5 inhibited adhesion of HL-60 cells to P- and E selectin-producing COS cells in vitro and immunoprecipitated a P-selectin glycoprotein ligand-1-like glycoprotein from HL-60 cell lysate, suggesting that it recognizes a functional consensus structure on the ligands for all selectins. 2H5 reacted not only with human but also with rat and mouse neutrophils. 2H5 is the first antibody against SLex that recognizes neutrophils of nonhuman mammals. The carbohydrate structure recognized by 2H5 was present not only on high endothelial venules of rat lymphoid organs but also on the endothelial cells of nonlymphoid organs. Furthermore, administration of the antibody markedly inhibited L- and P-selectin-mediated neutrophil rolling and adhesion in rat mesenteric venules in vivo. These results provide evidence for the presence of a consensus carbohydrate structure on the ligands for all selectins. The consensus structure thus has the potential to serve as a therapeutic target. PMID- 7485560 TI - Abnormalities of K+ and Ca2+ currents in ventricular myocytes from rats with chronic diabetes. AB - Ionic mechanisms related to the prolongation of cardiac action potential in rats with chronic diabetes mellitus were studied using whole cell voltage-clamp techniques. Diabetes was induced by injection of streptozotocin (STZ; 65 mg/kg body wt) into the tail vein, and ventricular myocytes were isolated from STZ injected rats (24-30 wk) and from age-matched normal rats. The current densities of transient outward current (Ito), a steady-state outward current, and L-type Ca2+ current (ICa) were significantly smaller in cells from diabetic animals. In addition, the kinetics of Ito of diabetic cells were modified. 1) The decay of Ito was well fitted by a sum of two exponential components in normal cells; there was only one (slow) component in the diabetic cells. 2) The steady-state inactivation curve of Ito in diabetic cells shifted by 5 mV in the negative direction. 3) Recovery from inactivation of Ito was slower in cells from diabetic animals. These alterations in Ito and the steady-state outward current can account for most of the action potential prolongation heretofore documented. The decrease of ICa may possibly be related to the depressed contraction seen in chronic diabetic mellitus. PMID- 7485561 TI - Acute effects of glucose on reactivity of cerebral microcirculation: role of activation of protein kinase C. AB - Our first goal was to determine whether acute hyperglycemia alters endothelium dependent reactivity of rat cerebral arterioles. Our second goal was to investigate a possible mechanism for impaired reactivity during acute hyperglycemia. Diameter of pial arterioles was measured during suffusion with ADP, acetylcholine, histamine, N-methyl-D-aspartate (NMDA), and nitroglycerin before and during application of a suffusate containing D-glucose (5, 10, 20, and 25 mM). ADP, acetylcholine, histamine, NMDA, and nitroglycerin produced dose related vasodilation before application of D-glucose. Vasodilatation in response to the agonists was not altered by 5 and 10 mM D-glucose. In contrast, vasodilatation in response to ADP, acetylcholine, histamine, and NMDA was impaired during application of 20 and 25 mM D-glucose. Dilatation in response to nitroglycerin was not altered. Application of the protein kinase C inhibitor calphostin C (1.0 nM) or chelerythrine (10 nM) restored endothelium-dependent vasodilatation during application of 25 mM D-glucose. Thus acute hyperglycemia impairs endothelium-dependent responses of cerebral arterioles via the activation of protein kinase C. PMID- 7485564 TI - Biaxial mechanical behavior of excised porcine mitral valve leaflets. AB - Anterior and posterior leaflets from excised porcine mitral valves were mechanically tested under cyclic equibiaxial and strip biaxial stretch protocols at a strain rate of 4-12%/s after preconditioning. Cauchy stress and Lagrangian strain were calculated for both membrane and three-dimensional cases. The leaflets exhibited nonlinearly elastic, anisotropic behavior. Both anterior and posterior leaflets were less extensible in the circumferential than in the radial direction under equibiaxial stretch, with stress ratios of 5.7 and 4.3, respectively. The posterior leaflets exhibited greater extensibility in both directions and lower circumferential posttransitional moduli (ranges 690-820 vs. 2,500-3,200 N/m for anterior). This larger posterior extensibility may be due to the greater number of chordal attachments, which provide additional mechanical stability to this structure. Coupling of radial and circumferential mechanical behavior was evidenced by the response to different stretch protocols, indicating a complex microstructural coupling between individual collagen fibers or bundles. These are the first biaxial data for mitral valves and are a foundation for the development of a more detailed quantitative material description. PMID- 7485563 TI - Impaired arginine metabolism and NO synthesis in coronary endothelial cells of the spontaneously diabetic BB rat. AB - Arginine metabolism via nitric oxide (NO) synthase and other pathways was studied in coronary endothelial cells (EC) from the spontaneously diabetic BB rat, an animal model of human type I diabetes mellitus (IDDM). EC were prepared from insulin-treated diabetic BB (BBd) and non-diabetes-prone BB (BBn) rats. Basal NO synthesis was studied in EC cultured for 48 h in medium containing 0.4 mM L arginine. At the end of the culture period, the medium was analyzed for nitrite and nitrate (two major end stable oxidation products of NO), and the cells were used to determine arginine uptake and metabolism and the activities of some arginine-degrading enzymes. For studies of arginine metabolism, cells were incubated at 37 degrees C for 1 h in Krebs-Henseleit bicarbonate buffer (pH 7.4) containing 1 mM L(-)[1-14C]arginine or L(-)[1-14C]ornithine. The rates of production of nitrite plus nitrate by BBd EC were only 15% of those of BBn cells. This impaired NO synthesis in BBd EC was not due to alterations in arginine uptake, NO synthase activity, or intracellular arginine concentrations but might have resulted from a limited intracellular availability of cofactors of NO synthase. In addition to the arginine-NO pathway, arginine was found to be metabolized to urea, ornithine, and, to a much lesser extent, CO2 via arginase and ornithine aminotransferase. The activities of arginase and the formation of ornithine and urea from arginine were decreased by 90% in BBd compared with BBn cells. These results, coupled with the reduced NO synthesis, indicate metabolic defects in arginine metabolism in BBd EC.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7485565 TI - Determination of myoglobin saturation of frozen specimens using a reflecting cryospectrophotometer. AB - This report describes a method and instrumentation for determining myoglobin (Mb) oxygen saturation in skeletal muscle. Canine gracilis is frozen in situ using a liquid N2-cooled copper block. Transverse section surfaces of frozen unstained muscle are observed at -110 degrees C using a microspectrophotometric system. The Mb saturation is determined using epi-illumination and a four-wavelength optical method. A special aperture permits illumination of a 20-microns-square area, and the radius of the catchment volume is estimated to be approximately 60 microns, with the strongest signal arising from the central region. The equibestic wavelengths used were 546.6, 570.5, and 584.1 nm. The method was validated using the nonlinear multicomponent analysis method of Lubbers. End-point (0 and 100% saturation) calibration was set using ischemic and adenosine-treated highly oxygenated muscles, respectively. The effects of hemoglobin (Hb) and metmyoglobin (metMb) signal contamination were evaluated experimentally and by computer-mixing simulations. Mb saturation determinations adjacent to large vessels are to be avoided. MetMb and capillary Hb do not interfere with the determination. The reproducibility of the method is estimated to be +/- 5%. PMID- 7485562 TI - Various cells release a stable small molecule that inhibits endothelium-dependent relaxation. AB - Previous studies have shown that neutrophils release a stable factor that inhibits endothelium-dependent relaxation. In the present studies, the effects of supernatants derived from various cells on endothelium-dependent relaxation were studied. Cells were obtained from seven sources: human hematopoietic cells including mononuclear leukocytes (MONO), polymorphonuclear leukocytes (PMNs), and chronic lymphocytic leukemia (CLL) cells; cells of the cardiovascular system including human endothelial cell line ECV304, human smooth muscle cells, and rat myocardial cells; and the tumor cell line HPB. These isolated or cultured cells were incubated for 1 h in Krebs solution to release the factor. The results showed that the supernatants from 10(5) cells/ml of all cells except the tumor cell line HPB produced a potent inhibitory effect on endothelium-dependent relaxation of rat aortic rings in response to acetylcholine and Ca2+ ionophores A23187 and ionomycin but not on endothelium-independent relaxation to nitroprusside and glyceryl trinitrate. When the concentration increased to 10(6) cell/ml, the supernatants from the tumor cell line HPB also slightly but significantly inhibited endothelium-dependent relaxation. The potency order was PMNs = MONO = CLL cells > cardiac cells > smooth muscle cells > the endothelial cell line ECV304 > the tumor cell line HPB. It seems that the hematopoietic cells and the cardiac cells are more active in release of the factor. The effect of this factor was rapid in onset and hard to wash out. A cyclooxygenase inhibitor or a thromboxane A2-prostaglandin H2 receptor antagonist partially but significantly reduced the effect of the factor.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7485566 TI - Resting oxygenation of rat and rabbit intestine: arteriolar and capillary contributions. AB - Counter-current exchange of oxygen may occur between inflow and outflow microvessels of the small intestine and greatly influence the dominant sites of tissue oxygenation. To determine the location and magnitude of potential exchange, percent saturation of hemoglobin with oxygen (%SHb) was measured in microvessels throughout the intestine of rats and rabbits. Oxygen losses from systemic arterial blood through large and intermediate arterioles (second order, 2A) was 5-7%SHb in both species, and there was no evidence of an increase in percent saturation along intermediate and large venules. A larger loss of oxygen from arterioles and an increase in venous saturation would be evident if significant arteriolar to venular counter-current exchange of oxygen occurred in the submucosa. From 2A to the villus tip, arteriolar saturation decreased approximately 10%SHb in rabbits and approximately 15%SHb in rats; the villus tip percent saturation was 72.9 +/- 3.9%SHb in rabbits and 69.9 +/- 2.9%SHb in rats. An additional decrease of 5%SHb in rabbits and 15%SHb in rats occurred across the villus capillaries and smallest venules. Although the total reduction in percent saturation across the villi was different between the two species, 70-90% of the total arteriovenous oxygen losses occurred in the capillaries and small arterioles of the villi. We found no evidence of counter-current exchange of oxygen in villi or any other vascular region. Rather, as appears to occur in most organs, small arterioles in conjunction with capillaries dominate resting oxygen exchange to tissue. PMID- 7485567 TI - Different properties of the atrial G protein-gated K+ channels activated by extracellular ATP and adenosine. AB - Extracellular ATP (ATPo) and adenosine activate G protein-gated inwardly rectifying K+ currents in atrial cells. Earlier studies have suggested that the two agonists may use separate pathways to activate the K+ current. Therefore, we examined whether the K+ channels activated by the two agonists have different properties under identical ionic conditions. In cell-attached patches, K+ channels activated by 100 microM ATP in the pipette had a single-channel conductance and mean open time of 32.0 +/- 0.2 pS and 0.5 +/- 0.1 ms, respectively, compared with 31.3 +/- 0.3 pS and 0.9 +/- 0.1 ms for the K+ channels activated by adenosine (140 mM KCl). With ATPo as the agonist, the K+ channel activity in cell-attached patches was approximately threefold lower than that in inside-out patches with 100 microM GTP in the bath. Applying ATP to the cytoplasmic side of the membrane (ATPi) produced a biphasic concentration dependent effect on channel activity: an increase at low [mean affinity constant (K0.5) = 190 microM] and a decrease at high (K0.5 = 1.3 mM) concentrations. In contrast, with adenosine as the agonist, K+ channel activity in cell-attached patches was approximately fourfold greater than that in inside-out patches with 100 microM GTP in the bath. In inside-out patches, ATPi only augmented the K+ channel activity (K0.5 = 32 microM). These results show that although both ATPo and adenosine activate kinetically similar K+ channels in atrial cells, the channels are regulated differently by intracellular nucleotides. PMID- 7485568 TI - Relationship of simultaneous atrial and ventricular pressures in stage 16-27 chick embryos. AB - Ventricular filling is determined by a dynamic balance between atrial and ventricular load and function. The embryonic cardiovascular system undergoes simultaneous growth and morphogenesis at the cellular, tissue, and organ levels to match the embryo's geometrically increasing metabolic demands. As part of our long-term investigation of atrial/ventricular coupling during primary cardiac morphogenesis, we defined the relationship between simultaneous atrial and ventricular pressures in the stage 16-27 white Leghorn chick embryo. We measured atrial and ventricular blood pressures with servo-null micropressure systems and sampled analog waveforms digitally at 500 Hz. Peak atrial pressure increased geometrically from 0.38 +/- 0.03 to 1.21 +/- 0.17 mmHg, while ventricular end diastolic pressure increased linearly from 0.18 +/- 0.03 to 0.55 +/- 0.04 mmHg. The passive and active mean pressure gradients increased from 0.23 +/- 0.04 and 0.20 +/- 0.03 mmHg at stage 16 to 0.52 +/- 0.10 and 0.62 +/- 0.11 mmHg at stage 27, respectively. The atrioventricular pressure gradients were similar for stages 16, 18, and 21, then increased to stage 27. This diastolic pressure gradient identifies the atrioventricular orifice and developing endocardial cushions as a site of flow resistance that may influence both ventricular filling and chamber morphogenesis. PMID- 7485569 TI - Pressure wave propagation in a multibranched model of the human upper limb. AB - The influence of the large arteries and the peripheral load on pressure wave propagation in the human upper limb was investigated in an anatomically realistic multibranched model based on linear transmission theory. To mimic vascular changes seen in life, the viscoelastic properties of large arteries and the peripheral load properties (represented as modified windkessels) were altered as follows: Young's modulus (from 10.9 x 10(6) to 15.3 x 10(6) dyn/cm2) and phase (from 0 to 15 degrees) of the complex elastance, windkessel time constant (from 0 to 0.6 s), and peripheral reflection coefficient (from 0 to 0.95). The relationship between the central aortic and peripheral radial pressure waveforms was analyzed in the time and the frequency domain. Results indicate that the large arterial properties have less influence (peak systolic pressure changed by 3% and peak of transfer function changed by 29%) than the properties of the peripheral load (systolic pressure changed by 14% and peak of transfer function changed by 74%) on the pressure wave propagation in the upper limb. PMID- 7485570 TI - Role of bradykinin in cardiac functional protection after global ischemia reperfusion in rat heart. AB - We have reported that cardiac preconditioning against ischemia-reperfusion (IR) can be induced by transient ischemia (TI) and alpha 1-adrenoreceptor stimulation, both mediated by protein kinase C (PKC) (Mitchell, M., X. Meng, C. Parker, E. Brew, A. Harken, and A. Banerjee. Circ. Res. 76: 73-81, 1995). Our study objective was to explore the mechanism of endogenous preconditioning and address the role of PKC activation in bradykinin-mediated cardiac functional protection. Isolated rat heart was used to assess the effects of exogenous bradykinin, TI, selective B2-receptor blocker, and PKC antagonism on cardiac functional recovery after a global IR injury. Final recovery of developed pressure was improved in hearts treated with bradykinin and TI compared with controls. Bradykinin- and TI mediated preconditioning was eliminated with coinfusion of the B2-receptor antagonist. Further evaluation of bradykinin-mediated preconditioning revealed that PKC blockade also eliminated functional protection. Immunofluorescent stains of bradykinin-treated hearts demonstrated translocation and activation of specific PKC isoforms in the preconditioned heart. We conclude that TI-mediated preconditioning involves intrinsic cardiac bradykinin receptor stimulation. Bradykinin, through the B2 receptor, initiates a series of intracellular events culminating in the activation of PKC. PMID- 7485572 TI - Prolonged leukocyte transit time in coronary microcirculation of endotoxemic pigs. AB - We quantified the timing and extent of leukocyte retention by the coronary microcirculation in a pig model of hyperdynamic sepsis in three ways. First, the transmyocardial leukocyte gradient was determined as coronary blood flow (calibrated ultrasonic flow probe) multiplied by the difference between leukocyte counts in the aorta and coronary sinus. Measurements were taken at 1-min intervals for 30 min and then at 3-min intervals for 45 min in anesthetized pigs exposed to either endotoxin (50 micrograms/kg iv over 30 min) (n = 7) or vehicle (n = 7). Second, postmortem morphometric analysis was used to quantitate the number and location of retained myocardial leukocytes. Finally, myocardial capillary transit time of leukocytes was calculated from the above measures. In the endotoxin group 2.1 +/- 0.8 x 10(9) leukocytes/100 g wet wt were retained in the coronary circulation, primarily in capillaries. This resulted in 111 +/- 37 (P < 0.05) times as many leukocytes in the coronary microcirculation than predicted from the arterial leukocyte concentration. Myocardial capillary transit time of leukocytes was prolonged to 39.1 +/- 20.6 s (P < 0.05) in the endotoxin group versus 5.0 +/- 1.4 s in the control group. We conclude that, after endotoxin infusion in a pig model of hyperdynamic sepsis, myocardial leukocyte transit is slowed, leading to the retention of large numbers of leukocytes in the coronary microcirculation. PMID- 7485571 TI - Role of NO mechanism in cardiovascular effects of diaspirin cross-linked hemoglobin in anesthetized rats. AB - The role of nitric oxide (NO) in the cardiovascular actions of diaspirin cross linked hemoglobin (DCLHb) was studied in anesthetized rats. The regional circulatory and systemic hemodynamic effects of DCLHb (400 mg/kg iv) were studied using a radioactive microsphere technique in control (untreated) and L-arginine (a NO precursor) pretreated rats. DCLHb produced a significant increase in blood pressure (75%), cardiac output (42%), stroke volume (36%), and total peripheral resistance (45%), without affecting heart rate, when administered to control rats. L-Arginine pretreatment significantly attenuated DCLHb-induced systemic hemodynamic effects. DCLHb-induced increase in blood flow to the skin and spleen was completely blocked, and that to the heart was partially blocked, by L arginine pretreatment, suggesting that cardiovascular actions induced by DCLHb could be antagonized by the NO precursor L-arginine. The NO synthase (NOS) inhibitor NG-nitro-L-arginine methyl ester (L-NAME) produced significant increases in regional vascular resistance, leading to a decrease in blood flow to all the organs except the heart, where an increase in blood flow and a decrease in vascular resistance was observed. DCLHb, when administered in L-NAME pretreated rats, accentuated the decrease in blood flow to the gastrointestinal system, spleen, mesentery and pancreas, skin, and musculoskeletal system. These studies provide evidence that the NO precursor L-arginine can attenuate the effects of DCLHb and that DCLHb can potentiate the effect of the NOS inhibitor L NAME. The role of NO in the mechanism of action of DCLHb was further studied by estimating plasma guanosine 3',5'-cyclic monophosphate (cGMP) in control, DCLHb treated, L-NAME-treated, and L-NAME followed by DCLHb-treated rats. DCLHb and L NAME significantly decreased the concentration of circulating cGMP in blood plasma. L-NAME pretreatment potentiated DCLHb-induced decrease in cGMP levels. Because the formation of cGMP is stimulated by NO, these studies provide additional evidence for the involvement of NO in the mechanism of action of DCLHb. It is concluded that NO plays an important role in the cardiovascular effects of DCLHb. PMID- 7485573 TI - Adhesion of flowing neutrophils to cultured endothelial cells after hypoxia and reoxygenation in vitro. AB - Using a novel in-line deoxygenating system linked to an in vitro flow-based adhesion assay and video microscopy, we have studied neutrophil recruitment and migration after hypoxia and reoxygenation of cultured human umbilical vein endothelial cells (HUVEC). Unstimulated purified neutrophils were perfused over reoxygenating HUVEC immediately after various periods of endothelial hypoxia. Adhesion to HUVEC was dependent on the duration of hypoxia, with 30, 60, and 100 min of exposure causing graded increments in neutrophil recruitment. The degree of hypoxia also markedly influenced the endothelial response. Severe hypoxia (O2 < 2.5%) induced stationary attachment and then migration of neutrophils, in contrast to rolling adhesion alone under a less intense regime (O2 = 2.5-4.0%). Judged from studies with monoclonal antibodies, P-selectin was essential for adhesion after severe hypoxia, and neutrophil immobilization was attributable to the activation of neutrophil beta 2-integrin. Perfusion of neutrophils with an antibody against interleukin-8 or a platelet-activating factor antagonist reduced levels of adhesion. However, IL-8 appeared to be the dominant agent involved in the immobilization from flow, whereas platelet-activating factor was the more potent agent involved in initiating subendothelial migration. Thus endothelial cells alone can initiate all stages of adhesion and migration of flowing neutrophils after hypoxia and reperfusion. PMID- 7485574 TI - Flow-dependent arteriolar dilation in normotensive rats fed low- or high-salt diets. AB - Ingestion of a high-salt diet has previously been shown to suppress the endogenous influence of nitric oxide (NO) on arteriolar tone in hypertension resistant, salt-resistant Dahl (SR/Jr) rats. Because luminal blood flow can be an important stimulus for endothelial NO release, this study was undertaken to determine whether high salt intake can also lead to a deficit in the direct flow dependent regulation of arteriolar diameter. The spinotrapezius muscle microvasculature was studied by in vivo microscopy in SR/Jr rats fed low (0.45%)- or high (7%)-salt diets for 2 wk, and arcade arteriole responses to increased luminal flow (via parallel vessel occlusion) were studied in both dietary groups. There was no significant difference between groups in arterial pressure or in resting arteriolar diameters, volume flows, or wall shear rates. In low-salt SR/Jr, a 36% increase in luminal flow produced an average arteriolar dilation of 38% that was significantly reduced by the NO synthase inhibitor NG-monomethyl-L arginine (L-NMMA). In high-salt SR/Jr, a similar flow increase produced an average dilation of only 16% (P < 0.05 vs. low-salt SR/Jr), and this response was unaffected by L-NMMA. Inhibition of cyclooxygenase activity with meclofenamate had no effect on this response in either group. These findings suggest that NO release mediates a portion of flow-dependent arteriolar dilation in rat spinotrapezius muscle and that high salt intake, in the absence of hypertension, can attenuate this response via a suppression of NO activity. PMID- 7485575 TI - Preconditioning accelerates contracture and ATP depletion in blood-perfused rat hearts. AB - We investigated the effect of preconditioning on the ischemia-induced depletion of ATP in the blood-perfused rat heart. Isolated hearts (n = 5/group) were aerobically perfused with whole blood from a support rat and subjected to zero flow global ischemia (37 degrees C) for periods up to 35 min. Frozen hearts were taken for metabolic analysis. Ischemic contracture was assessed with an isovolumic intraventricular balloon. The study groups were 1) control (C) with unprotected ischemia, 2) preconditioning (PC; 2 cycles of 3-min ischemia/3-min reperfusion), and 3) cardioplegia (CP; St. Thomas') before ischemia. Preconditioning accelerated, whereas cardioplegia delayed, ischemic contracture (time to peak contracture: PC = 8.1 +/- 0.3 and CP = 25.1 +/- 0.2 min vs. C = 15.6 +/- 0.3 min, P < 0.05). The ischemia-induced decline in ATP was delayed by cardioplegia but accelerated by preconditioning (P < 0.05). In a parallel study, preconditioning and cardioplegia protected postischemic contractile function to a similar extent. Thus, in the blood-perfused rat heart, preconditioning accelerated ischemic contracture and depletion of ATP. In contrast, cardioplegia slowed ischemic contracture and ATP depletion. PMID- 7485576 TI - NMDA receptors in NTS are involved in bradycardic but not in pressor response of chemoreflex. AB - Activation of carotid chemoreceptors with intravenous potassium cyanide (KCN) produces increases in arterial pressure, bradycardia, and tachypnea. In the present study, we activated carotid chemoreceptors with KCN and the neurotransmission of the chemoreceptor reflex into the commissural nucleus tractus solitarii (NTS) was blocked with phosphonovaleric acid (AP-5), an N methyl-D-aspartate (NMDA)-selective antagonist. The aim of this study was to evaluate the involvement of NMDA receptors in the cardiovascular and respiratory responses produced by chemoreceptor activation in unanesthetized rats. The pressor response to KCN was not changed after microinjection of three different doses of AP-5 into the NTS, whereas the bradycardic response was reduced in a dose-dependent manner. The increase in respiratory frequency in response to carotid chemoreceptor activation was also not affected by AP-5 microinjected into the NTS. The data indicate that the activation of the cardiovagal component of the chemoreflex in the commissural NTS is mediated by NMDA receptors, whereas pressor and ventilatory responses are not. PMID- 7485577 TI - Muscle metaboreceptors in hemodynamic, autonomic, and ventilatory responses to exercise in men. AB - A muscle metaboreceptor (ergoreceptor) contribution to the hemodynamic and autonomic responses to exercise is well recognized, but a ventilatory component remains controversial. Control handgrips were compared with handgrips followed by 4-min regional circulatory occlusion of the exercising muscles to isolate the metaboreceptor role in blood pressure, autonomic tone (spectral analysis of R-R and blood pressure variability), and ventilatory responses to exercise in 11 normal subjects. Exercise responses were maintained after the effort by metaboreflex activation in systolic pressure (136.2 +/- 3.5 vs. 123.0 +/- 4.3 mmHg, P < 0.05), ventilation (19.0 +/- 2.6 vs. 8.5 +/- 0.4 l/min, P < 0.0005), and sympathetic discharge to the heart and circulation (elevated low-frequency components of R-R interval, 1,747.5 +/- 309.2 vs. 1,085.9 +/- 259.1 ms2, P < 0.05, and systolic pressure variability, 45.3 +/- 3.9 vs. 26.5 +/- 4.4 mmHg2, P < 0.005). We conclude that metaboreflex contributes to the sympathetic, hypertensive, and hyperpneic responses to exercise in normal subjects. PMID- 7485578 TI - Influence of respiration on metabolic, hemodynamic, psychometric, and R-R interval power spectral parameters. AB - Because respiration modulates autonomic activity, we determined the magnitude of perturbation of changing breathing frequency and tidal volume on metabolic, hemodynamic, psychometric, and R-R interval power spectral parameters. Seated subjects breathed at three different rates and five different volumes with each of the different rates. Breathing rates and volumes were percentages of the subject's resting breathing pattern and, therefore, identical across all subjects. Increases in rate and volume resulted in significant perturbations in end-tidal CO2 production, CO2 production, ventilatory equivalent for O2, comfort levels, and R-R interval power spectra (P < 0.05). The magnitude of the perturbations in the above parameters indicated a substantial upset in all subjects' metabolic, hemodynamic, and comfort homeostasis, precipitating a significant loss of vagal tone. The implications of our findings are that imposed breathing patterns used to modulate autonomic outflow should be tailored to the individual's resting breathing pattern. These data further support the urgent need for concomitant metabolic and respiratory measurements when analyzing and interpreting heart rate variability data. PMID- 7485579 TI - Activation delay in healed myocardial infarction: a comparison between model and experiment. AB - Conduction delay in healed myocardial infarction, facilitating reentry, is frequently based on an increased path length the activation has to travel in a matrix of merging and diverging bundles that survive in the infarcted area. Additional delay occurs at sites where bundles bifurcate. The purpose of this study was to investigate conduction delay at sites where bundles bifurcate. A computer model was developed to simulate spread of activation in a two dimensional sheet of excitable elements. A structure consisting of two isolated bundles merging into a single one was modeled. Extracellular electrograms calculated in the model were comparable to electrograms obtained in a superfused infarcted papillary muscle model. A zone of crowded isochrones or local conduction delay was found at the site where an isolated bundle bifurcated. The position of the isochrones in this area depended on the way activation times were determined. Lines of activation delay were mainly perpendicular to the fiber direction. In conclusion, the results have enabled us to better understand extracellular electrograms at pivoting points and show that activation sequences at a microscopic level can best be constructed on the basis of Laplacian signals. PMID- 7485580 TI - Effects of indomethacin on brain blood flow, cerebral metabolism, and sagittal sinus prostanoids after hypoxia. AB - We tested the hypotheses that during recovery from hypoxia, newborn piglets exhibit a posthypoxic cerebral hyperemia, indomethacin-pretreated piglets exhibit a posthypoxic cerebral hypoperfusion, and that the changes caused by indomethacin are dose dependent and related to the loss of prostanoids. We studied piglets exposed to 40 min of hypoxia after pretreatment with high (5 mg/kg, n = 9) or low (0.3 mg/kg, n = 8) doses of indomethacin or placebo (n = 9) and allowed to recover for 120 min. In the placebo and low-dose pretreatment groups, total and regional brain blood flow increased during hypoxia but returned to baseline 10 min after hypoxia. High-dose indomethacin pretreatment was associated with a posthypoxic hypoperfusion to certain brain regions at 10 min of recovery to values similar to those after indomethacin treatment before the onset of hypoxia. During and after hypoxia, the cerebral metabolic rate of oxygen was preserved in both the placebo and low-dose groups and decreased significantly during hypoxia in the high-dose group. Sagittal sinus prostacyclin was reduced significantly in both indomethacin-treated groups throughout the study. We conclude that a posthypoxic hyperemia is not observed in newborn piglets. This finding was not altered by pretreatment with a therapeutic dose of indomethacin, whereas a pharmacological dose was associated with selective hypoperfusion to certain brain regions both before hypoxia and during recovery from hypoxia. PMID- 7485581 TI - Effects of ischemic and adenosine preconditioning on interstitial fluid adenosine and myocardial infarct size. AB - The accumulation of adenosine during a brief coronary occlusion has been proposed to mediate the infarct size-limiting effect of ischemic preconditioning. The purpose of this study was to compare the effects of ischemic preconditioning and a transient adenosine infusion on myocardial interstitial fluid (ISF) adenosine levels and infarct size. Microdialysis fibers (10.0 mm length) were placed in the left ventricular myocardium of pentobarbital sodium-anesthetized rabbits to estimate ISF adenosine. Ischemic preconditioning was induced by 5 min of coronary artery occlusion and 10 min of reperfusion before 45 min of occlusion. Adenosine preconditioning was induced with 5 min of intravenous adenosine infusion (140 micrograms.kg-1.min-1) followed by a 10-min washout before the prolonged occlusion. Myocardial infarct size was determined by triphenyltetrazolium chloride staining after 3 h of reperfusion. Five minutes of ischemia and 5 min of adenosine infusion produced comparable increases in dialysate adenosine levels (from 0.19 +/- 0.02 to 0.69 +/0- 0.11 and 0.28 +/- 0.10 to 0.71 +/- 0.18 microM, respectively) that decreased to baseline before the prolonged ischemia; however, ischemic-preconditioned hearts exhibited elevated dialysate adenosine levels for the first 5 min of reperfusion. Ischemic-preconditioned hearts exhibited significantly reduced dialysate adenosine concentrations for the first 20 min of the prolonged occlusion (P < 0.05 vs. control), and infarct size was reduced from 41 +/- 6 to 10 +/- 4% of risk area. Adenosine preconditioning had no effect on dialysate adenosine levels during prolonged ischemia but did reduce infarct size to 25 +/- 5% of risk area. These results indicate that a transient increase in ISF adenosine can reduce myocardial infarct size, but adenosine alone does not fully replicate the protective effects of ischemic preconditioning. PMID- 7485582 TI - Vasoactive intestinal polypeptide antagonists attenuate vagally induced tachycardia in the anesthetized dog. AB - We used three vasoactive intestinal polypeptide (VIP) antagonists, VIP-(10-28), [p-Cl-D-Phe6,Leu17]VIP, and NT-VIP, to evaluate the role of VIP as a mediator of vagally induced tachycardia in chloralose-anesthetized dogs. After we administered muscarinic and beta-adrenergic receptor antagonists, we evoked vagally induced tachycardia either directly, by stimulating the vagus nerves for 2 min, or reflexly, by injecting phenylephrine to increase blood pressure. Furthermore, each of the antagonists attenuated the tachycardias induced by vagal stimulation by approximately 50% and the reflexly induced tachycardias by approximately 70%. Each VIP antagonist attenuated the chronotropic responses that we evoked by injecting VIP (5.2 ng/kg) into the sinus node artery. We tested the specificity of these VIP antagonists by determining whether they attenuated the increases in heart rate evoked by two other neuropeptides [peptide histidine isoleucine (PHI) and glucagon]. VIP-(10-28) attenuated the response to PHI, but not to glucagon. The other two VIP antagonists did not alter the chronotropic responses to PHI or glucagon. Our results support the hypothesis that neurally released VIP is the principal mediator of vagally induced tachycardia in the dog. PMID- 7485584 TI - Gap junction function in vascular smooth muscle: influence of serotonin. AB - In this study we examined the effects of serotonin (5-hydroxytryptamine, 5-HT) on the function of gap junctions between smooth muscle cells isolated from human and pig coronary and rat mesentery arteries and between A7r5 cells (cell line derived from embryonic rat aorta). Mesentery and pig coronary cells expressed connexin (Cx) 43, and human coronary cells expressed Cx40. Mesentery and pig coronary cells each exhibited a single gap junction channel population with unitary conductances of 75 and 59 pS, respectively. Human coronary cells exhibited two channel populations with unitary conductances of 51 and 107 pS. The A7r5 cells express Cx40 and Cx43 and exhibit three channel populations with unitary conductances of 70, 108, and 141 pS. Under control conditions, junctional conductance between the four cell types ranged from 11 to 20 nS. During maximal stimulation with 5-HT (1-10 microM), junctional conductance increased (29-75%) in all four cell types. The unitary conductance profiles in the rat mesentery and pig coronary cells were unaffected by 5-HT, suggesting that the observed increase in macroscopic conductance reflects an increase in open probability. Unitary conductances were also unaffected in the human coronary and A7r5 cells. However, there was a reduced frequency of the 105-pS channel in the human coronary cells and of the 70- and 141-pS channels in the A7r5 cells. These changes in the relative frequency histograms suggest that the open probabilities of the various channel types are differentially affected by the 5-HT treatment.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7485583 TI - Nature of heart failure in patients with ventricular septal defect. AB - To assess the contributions of systolic and diastolic dysfunction to congestive heart failure (CHF) in ventricular septal defect (VSD), we studied 13 children with VSD at catheterization using a Millar catheter. Eight children had CHF, whereas five did not. Phenylephrine was infused at a rate of 5 micrograms.kg 1.min-1, and M-mode echocardiography and pressure were measured simultaneously. Systolic left ventricular (LV) function was assessed by maximum LV pressure (LVP), rate of pressure development (dP/dt), and by the end-systolic pressure diameter relation (ESPDR). Systolic myocardial function was assessed by the end systolic stress-strain relation. Diastolic chamber function was assessed by the isovolumic relaxation time constant (tau) and by the end-diastolic pressure diameter relation (EDPDR). Diastolic myocardial function was measured by the end diastolic stress-strain relationship. With phenylephrine, maximum LVP increased from 99 +/- 5 to 119 +/- 4 mmHg with CHF and from 106 +/- 6 to 149 +/- 10 mmHg without CHF. +dP/dt was lower with CHF (1,582 +/- 96 mmHg/s) than without CHF (2,300 +/- 200 mmHg/s). The maximum slope of the ESPDR was 39 +/- 8 with CHF and 94 +/- 14 mmHg/cm without CHF. The maximum slope of the midwall stress-strain relation was 223 +/- 37 with CHF and 395 +/- 93 g/cm2 without CHF. tau was 25 +/- 2 without CHF compared with 32 +/- 3 ms with CHF. The EDPDR was shifted leftward with failure, whereas the stress-strain relation was similar for all patients. CHF in patients with VSD results primarily from systolic dysfunction.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7485585 TI - Scatter in input impedance spectrum may result from the elastic nonlinearity of the arterial wall. AB - We have examined the role of the nonlinear elastic properties of the arterial wall on the human aortic input impedance obtained at different heart rates and different pressure and flow wave shapes. Pressure and flow were taken from a computer model that provides realistic simulations of the nonlinear distributed systemic arterial tree. Different wave shapes of ascending aorta pressure and flow and different heart rates were used to derive input impedance moduli and phase angles via Fourier analysis. The results show that the nonlinear elastic properties of the arterial wall are responsible for significant variations in the input impedance spectrum when changes in heart rate and aortic flow wave shape take place. This finding may explain the scatter often observed in experimentally determined input impedance data using different heart rates obtained by pacing. PMID- 7485586 TI - Laser-Doppler perfusion imaging of microvascular blood flow in rabbit tenuissimus muscle. AB - To evaluate a newly developed high-resolution laser-Doppler perfusion imager (HR LDPI) for analysis of local tissue perfusion, blood flow measurements in the rabbit tenuissimus muscle were carried out in combination with intravital microscopic observation. The principle of the LDPI method is based on a low-power laser beam scan of the exposed tissue from which a two-dimensional color-coded perfusion map is created through computerized signal analysis. The perfusion of the tenuissimus muscle prepared for microscopic observation was analyzed in a 5 mm x 8 mm area as the muscle was exposed to atmospheric oxygen tension (Po2; 20 kPa), a low Po2 (approximately 3 kPa), and after vasodilatation induced by topical application of prostaglandin E2 (PGE2). In selected areas free from larger vessels, a significantly lower perfusion average reading was demonstrated under high Po2 conditions compared with low Po2 conditions (P < 0.05, n = 5 animals), and application of PGE2 gave rise to an average reading significantly higher than that at low Po2 (P < 0.01, n = 6 animals). The results were in good agreement with the flow changes observed microscopically, and the architecture of the microvascular network, as depicted by in vivo micrographs, was clearly recognizable in the perfusion images. In conclusion, blood flow changes in the rabbit tenuissimus muscle induced by various stimuli were quantitated with the HR LDPI method and could be spatially resolved in great detail, illustrating the potential of using HR-LDPI for analysis of local blood flow and to reveal spatial perfusion heterogeneity in tissues. PMID- 7485588 TI - Multihormonal regulation of nephron epithelia: achieved through combinational mode? AB - The kidney is the main organ regulating composition of body fluids. A considerable number of hormones control the activity of renal cells to maintain hydromineral equilibrium. It becomes more and more difficult to interpret this multihormonal control in terms of regulatory processes. To illustrate this complexity, the hormonal regulation of electrolyte transport in the nephron thick ascending limb is taken as an example. This nephron segment is largely responsible for two kidney functions: the urinary-concentrating ability (by its capacity to deliver hypertonic sodium chloride into the medullary interstitium) and regulation of magnesium excretion into final urine. Six hormones are presently identified as acting on the transport of both sodium chloride and magnesium ions in this nephron segment. Therefore, the pertinent question is how the thick ascending limb and, hence, the kidney, is capable of regulating water balance independently from magnesium balance. It is proposed that the hormones act in combination: circulating levels of the individual hormones acting on these cells may determine the configuration of the paracellular and transcellular transport pathways of the epithelium either in the "sodium" or "magnesium" mode. The configuration would depend on the distribution and activity of the receptor at the surface of the basolateral membrane in contact with the circulating hormones. This distribution along with stimulation of respective signal transduction pathways would lead to the final biological effects. It is already known that the distribution of cell receptors may vary according to factors such as age, nutritional variability, hormonal status, degree of desensitization of the receptors, etc. The modulation of hormonal responses would depend therefore on the degree of coupling of hormone-receptor complexes to different intracellular transduction pathways and on the resulting negative and/or positive interactions between these pathways. PMID- 7485587 TI - Restoration of normal pH triggers ischemia-reperfusion injury in lung by Na+/H+ exchange activation. AB - The effects of acidotic extracellular pH and Na+/H+ exchange inhibition on ischemia-reperfusion (I/R)-induced microvascular injury were studied in the isolated, buffer-perfused rat lung. When lungs were subjected to 45 min of ischemia followed by 30 min of reperfusion, the capillary filtration coefficient (Kfc) increased significantly, resulting in a change in Kfc (delta Kfc) of 0.360 +/- 0.09 ml.min-1.cmH2O-1.100 g-1. Addition of hydrochloric acid to the perfusate before ischemia at a concentration sufficient to reduce perfusate pH from 7.38 +/ 0.03 to 7.09 +/- 0.04 completely prevented the increase in Kfc associated with I/R (delta Kfc = 0.014 +/- 0.034 ml.min-1.cmH2O-1.100 g-1). Addition of a Na+/H+ exchange inhibitor, 5-(N,N-dimethyl)-amiloride, to the perfusate either before ischemia or at reperfusion also prevented the I/R-induced permeability increase (delta Kfc = 0.01 +/- 0.02 and -0.001 +/- 0.02 ml.min-1.cmH2O-1.100 g-1, respectively). We conclude that restoration of flow at physiological pH to the postischemic lung activates the Na+/H+ exchange system, which may represent the "triggering mechanism" responsible for initiating reperfusion-induced microvascular injury. PMID- 7485589 TI - Blood volume restitution and growth in fetal lambs after acute hemorrhage. AB - The effects of 15-50% fetal hemorrhage (at approximately 1%/min) were studied in 13 pregnant ewes at 130-135 days of gestation for up to 5 days posthemorrhage. The upper limit of acute blood loss appears to be approximately 45%, and the rate of restoration of blood volume decreases with the severity of hemorrhage, particularly with hemorrhage > 30-40%. The restoration of fetal blood volume was due primarily to the restoration of plasma volume; in the animals subject to 40 45% blood loss (n = 9), red cell mass was still only 69.1 +/- 3.9% of the prehemorrhage value at day 5 posthemorrhage. There appear to be two phases in the restoration of plasma: 1) plasma volume was restored by 2 h posthemorrhage and 2) the restoration of plasma protein mass occurred primarily from 2 to 24 h. There was a significant correlation between blood volume and plasma protein mass. However, the regression line for the posthemorrhage days was shifted significantly upward in relation to that for the hemorrhage day because of a significant rise of plasma protein concentration. This may be important for the maintenance of blood volume after hemorrhage. Finally, fetal growth rate was determined by comparing fetal weight estimated in utero (from blood volume) with birth weight in 12 nonhemorrhaged control fetuses and in the 9 fetuses subject to 40-45% hemorrhage. The average rate of growth per day was 1.57 +/- 0.34% and 1.82 +/- 1.02%, respectively. The latter value is not significantly different from zero, suggesting that the acute blood loss impaired fetal growth. PMID- 7485590 TI - Oxygen consumption, acid-base status, and behavior during and after acute, severe hemorrhage in fetal lambs. AB - The metabolic and behavioral effects of 40-45% hemorrhage (at approximately 1%/min) were studied in nine fetal lambs in utero (130-135 days of gestation) until 2 days posthemorrhage. Umbilical blood flow (Qum) fell from 192 +/- 14 to 100 +/- 9 ml.min-1.kg-1 at the end of blood loss, and on the day of hemorrhage it was linearly related to blood volume. However, on the posthemorrhage days, Qum was restored even though blood volume was still reduced. Fetal O2 delivery (DO2) fell with the decrease in Qum and later due to anemia (decrease in hematocrit from 33.4 +/- 1.2 to 23.6 +/- 1.2%), from 946 +/- 81 to 494 +/- 47 mumol.min-1.kg 1, but the decrease was lessened because of a rise in umbilical venous PO2 (31.7 +/- 2.6 to 45.9 +/- 2.8 mmHg). Fetal O2 consumption was reduced during and for 2 h after hemorrhage (262 +/- 17 to 190 +/- 16 mumol.min-1.kg-1), and this was associated with modest lactic acidemia (1.25 +/- 0.11 to 2.91 +/- 0.43 mM). There was also a temporary reduction in fetal breathing activity, low voltage electrocortical state, and rapid eye movements, but this was not associated with hypoxemia. It is concluded that reductions in fetal DO2 achieved via fetal blood loss are better tolerated than during hypoxemia and that the associated depression in fetal biophysical activities involves mechanisms other than systemic hypoxia. PMID- 7485591 TI - Cold acclimation-recruited nonshivering thermogenesis: the Syrian hamster is not an exception. AB - Biochemical evidence from in vitro studies of brown adipose tissue in Syrian hamsters indicates a significant degree of recruitment of the tissue as an effect of cold acclimation. However, earlier in vivo studies indicate a lack of recruitment of nonshivering thermogenesis in the intact animal as a result of cold acclimation. Because of this apparent discrepancy, the occurrence of cold acclimation-recruited nonshivering thermogenesis in hamsters was investigated. Hamsters were cold acclimated to 6 degrees C or remained at 24 degrees C (controls), and their thermogenic response was investigated in an open-circuit system at 24 degrees C. Cold acclimation resulted in a small increase in resting metabolic rate and a major increase in the thermogenic response to norepinephrine (61% increase over resting metabolic rate in controls and 156% increase in cold acclimated animals). The absolute beta 3-specific adrenergic agonist CGP-12177 also induced a high rate of nonshivering thermogenesis, which was similarly recruited. It was concluded that, concerning the relative effect of recruitment on the capacity for nonshivering thermogenesis, the intact hamsters responded as would be predicted from in vitro experiments. Thus the hamster does not seem to constitute an exception to the general patterns described for other rodents concerning recruitment of nonshivering thermogenesis due to cold acclimation. PMID- 7485592 TI - Glomerular actions of arginine vasotocin in the in situ perfused trout kidney. AB - Recent measurements of plasma arginine vasotocin (AVT) in teleost fish suggest circulating concentrations of 10(-10)-10(-12)M. Previous studies of the renal actions of AVT in vivo suggest both diuretic and antidiuretic effects, but at unknown circulating concentrations. We have investigated the renal actions of 10( 9) and 10(-11) M AVT in vitro using an in situ perfused kidney preparation of rainbow trout (oncorhynchus mykiss). AVT increased vascular resistance (56%), reduced perfusate flow (P < 0.001), and increased interrenal aortic pressure (P < 0.001). AVT resulted in dose-dependent decreases in urine flow rates, glomerular filtration rates, and tubular transport maxima for glucose. AVT at 10(-11) M reduced relative free water clearances (P < 0.01), but urine/plasma inulin ratios were unchanged, whereas 10(-9)M AVT reduced urine/plasma inulin ratios (P < 0.01) and increased relative free water clearances (P < 0.05). The filtering population of glomeruli was reduced by both 10(-11) and 10(-9)M AVT to approximately one third of the glomeruli, and a similar population of arterially perfused but nonfiltering glomeruli emerged. These results demonstrate that physiological concentrations of AVT have potent glomerular antidiuretic action in the trout, reducing the number of functional glomeruli, and imply reduced individual nephron filtration rates. PMID- 7485593 TI - Lesion of central nucleus of amygdala promotes fat gain without preventing effect of exercise on energy balance. AB - Male Wistar rats with intact or lesioned central nucleus of amygdala (CeA) were kept at rest or subjected to a treadmill running program for 21 consecutive days. Food intake and body weight were monitored throughout the exercise training program. At the end of the program, rats were killed and their carcasses processed for analysis of the contents in energy, fat, and protein. Exercise and CeA lesions induced opposite effects on energy balance; exercise delayed gains in body energy and fat, whereas CeA lesions promoted them. Total energy intake was lower in exercised rats than in sedentary ones over the 12 and 24 h that followed exercise. Food intake was higher in lesioned rats than in intact animals over the second half of the 12-h period that followed exercise. There was no interaction effect of exercise and CeA lesions on energy balance and intake and on body composition. Plasma levels of adrenocorticotropin hormone and corticosterone were higher in exercised rats than in sedentary ones, but there was no difference between lesioned and intact rats. This study, as well as confirming the effect of exercise on energy balance, indicates that CeA lesions may promote energy deposition in rats. Above all the present results provide evidence that CeA does not represent a necessary neuroanatomic structure in the effect of exercise on energy balance. PMID- 7485594 TI - Central inhibitory action of peptide YY on gastric motility in rats. AB - Specific peptide YY (PYY) binding sites have recently been identified autoradiographically in the area postrema, nucleus of the solitary tract, and dorsal motor nucleus regions [collectively referred to as the dorsal vagal complex (DVC)] in rats. These medullary brain stem regions are responsible for vagovagal reflex control of gastric function, including gastric motility. We propose that PYY can modulate gastrointestinal functions, such as gastric motility, by interacting with PYY binding sites found in DVC. Furthermore, we predict that central PYY effects on gastric function are mediated by the vagus nerve. In the present study, urethan-anesthetized rats were used. PYY (20.0 and 2.0 fmol) injected directly into DVCs of the animals produced significant inhibition of gastric motility that was stimulated by centrally applied thyrotropin-releasing hormone (TRH). TRH is a well-accepted central stimulator of vagal efferent pathway to the stomach. Otherwise, an excitatory effect of PYY (2 pmol) on basal gastric motility was observed and considered as being pharmacological. The inhibitory effect of PYY was abolished completely by unilateral (the injection side) cervical vagotomy, suggesting that the inhibition was vagally dependent. These results support the view that physiological concentrations of PYY may inhibit proximal gut function as part of the "ileal brake" mechanism by acting directly on vagovagal control circuits in the dorsal medulla. However, extremely high doses of PYY may activate gastric function through pharmacological action at pancreatic polypeptide receptors in the DVC. PMID- 7485595 TI - Mechanisms of acid release in isolated gas gland cells of the European eel Anguilla anguilla. AB - Mechanisms of acid production and of acid release have been analyzed in isolated gas gland cells of the eel swimbladder using a cytosensor microphysiometer. Incubation of isolated cells with oxamic acid caused a dose-dependent decrease in the rate of proton release. At the highest oxamic acid concentration used (20 mmol/l), proton release was reduced by approximately 40%; incubation with sodium fluoride (10 mmol/l) or removal of glucose from the extracellular medium caused 60 and 80% reduction, respectively. NaCN had little effect on proton secretion. Proton release of isolated gas gland cells was largely dependent on the extracellular sodium concentration, and this sodium effect was in part inhibitable by amiloride. A 15-20% reduction in the rate of proton secretion was observed in the presence of 4,4'-diisothiocyanostilbene-2,2'-disulfonic acid, an inhibitor of anion exchange. Inhibition of mammalian H(+)-K(+) adenosinetriphosphatase with omeprazole had no effect, whereas bafilomycin, an inhibitor of vesicular H(+)-adenosinetriphosphatase, induced a 25% reduction in proton secretion. Ethoxzolamide, a membrane-permeable inhibitor of carbonic anhydrase, caused a 60% reduction in proton secretion (inhibition constant = 54.4 nmol/l). Prontosil-dextran, a membrane-impermeable sulfonamide, also reduced the proton release, thus indicating the presence of a membrane-bound carbonic anhydrase facing the extracellular space. PMID- 7485596 TI - Testicular regression and recrudescence without subsequent photorefractoriness in Siberian hamsters. AB - Adult male Siberian hamsters transferred from 16 h light/day (16L) to 14 h light/day (14L) underwent gonadal regression and recrudescence over the course of 24 wk; the duration of reproductive quiescence was shorter by 10 wk than in hamsters transferred from 16L to 10L. A decrease in day length (DL) at week 12 from 14L to 10L greatly extended the duration of testicular involution, whereas transfer at week 12 from 10L to 8L was without effect. Hamsters that had manifested gonadal regression and recrudescence in 14L immediately initiated a second regression when challenged with 10L. Intermediate DLs induce gonadal regression and recrudescence without rendering hamsters refractory to shorter DLs. Intermediate DLs sufficient to initiate gonadal regression may be too long to trigger the interval timer that eventually induces refractoriness to shorter DLs and may be successively interpreted as short and long days, respectively, by Siberian hamsters. PMID- 7485597 TI - Differential sympathetic nerve responses to nitric oxide synthase inhibition in anesthetized rats. AB - Recent studies have suggested that the interaction between the sympathetic nervous system and nitric oxide (NO) or nitrosyl factors may be an important means by which arterial blood pressure is regulated. We investigated whether NO synthase (NOS) inhibition modulates basal sympathetic nerve discharge (SND) in baroreceptor-innervated and -denervated, chloralose-anesthetized Sprague-Dawley rats. We recorded mean arterial pressure (MAP), renal SND, and lumbar SND before and after administration of the NOS inhibitor, NG-nitro-L-arginine methyl ester (L-NAME, 20 mg/kg iv). Two minutes after L-NAME administration in baroreceptor innervated rats, MAP increased (+23 +/- 3 mmHg), whereas renal (-45 +/- 6%, n = 7) and lumbar (-35 +/- 2%, n = 6) SND significantly decreased from control levels. These changes persisted for up to 20 min after L-NAME administration. In baroreceptor-denervated rats, L-NAME increased MAP (+40 +/- 6 mmHg) and decreased lumbar SND (n = 7) (-37 +/- 10% from control at 20 min post-L-NAME). In contrast, renal SND progressively increased (+33 +/- 8% at 20 min post-L-NAME) from control after L-NAME administration in baroreceptor-denervated rats (n = 7). These results demonstrate that NOS inhibition can produce nonuniform changes in SND in baroreceptor-denervated rats and suggest that endogenous nitrosyl factors provide tonic excitation to lumbar SND, whereas they provide a tonic restraint to renal SND. PMID- 7485598 TI - Lymphatic regulation of hematocrit during hypoxia in the toad Bufo woodhousei. AB - Hypoxia rapidly increases hematocrit (Hct) in anuran amphibians by reducing plasma volume, but the mechanism(s) mediating this response is unknown. We tested the hypothesis that, during hypoxia, plasma volume is reduced by impaired lymph heart (LH) function, decreasing lymph flow into the circulation. In Bufo woodhousei, we measured the effects of hypoxia on Hct, lymph heart rate (LHR), LH pressure, the movement of dye from the dorsal lymph sac to the arterial blood, and flow through an open LH cannula. We also tested whether splenic contraction or cholinergic nerves contribute to the hypoxia-induced changes. Graded hypoxia between 21 and 4% O2 produced graded increases in Hct (P < 0.0001) and decreases in LHR (P = 0.01). Hypoxia reduced the rate of increase in arterial Evans blue concentration after injection into the dorsal lymph sac (P = 0.041) and decreased flow through an open LH cannula (P < 0.012). Hypoxia increased Hct and reduced LHR similarly in control, splenectomized, and sham-splenectomized toads. Atropine had no significant effect on Hct and LHR. These results indicate that the LHs play a regulatory role in hypoxia-induced hemoconcentration. PMID- 7485599 TI - Intracellular pH in snake renal proximal tubules. AB - Intracellular pH (pHi) was studied in isolated proximal renal tubules of garter snakes (Thamnophis spp.) with oil-filled lumens under control conditions [N-2 hydroxyethylpiperazine-N'-2-ethanesulfonic acid (HEPES)-buffered medium with pH 7.4 at 25 degrees C] and in response to NH4Cl pulse. pHi was measured with the pH sensitive fluorescent dye 2',7'-bis(2-carboxyethyl)-5,6-carboxyfluorescein (BCECF). Control resting pHi (7.1) and acidification in response to NH4Cl pulse (minimum pHi, 6.6) were essentially the same in snake tubules with oil-filled lumens or perfused lumens and in rabbit S2 proximal tubules with oil-filled lumens. Rate of recovery of pHi (dpHi/dt) from acid to resting level in snake tubules (2.5 x 10(-3) pH U/s was about one-third of that in rabbit tubules. Resting pHi and dpHi/dt from acid to resting level were Na+ dependent in the distal portion but not the proximal portion of snake proximal tubules. However, dpHi/dt was not influenced by amiloride or 4,4'-diisothiocyanostilbene-2,2' disulfonic acid in snake proximal tubules, suggesting that the effect of Na+ on dpHi/dt and resting pHi may involve membrane potential. This study also indicates that oil-filled lumens do not interfere with measurements of resting pHi and do permit evaluation of pHi regulation at the basolateral membrane without complications from transport at the luminal membrane. PMID- 7485600 TI - On the fractal nature of heart rate variability in humans: effects of vagal blockade. AB - The purpose of the present study was to investigate the effects of the vagal blocker atropine on the fractal nature of human heart rate variability (HRV) at rest. Approximately 10-min segments of beat-to-beat heartbeat intervals, i.e., HRV, of 10 normal subjects and 11 cardiac disease patients were measured before and after intravenous injection of 0.5-0.75 mg atropine sulfate. HRV data were analyzed by coarse graining spectral analysis (Y. Yamamoto and R. L. Hughson, Physica 68D: 250-264, 1993) to break down their total power into harmonic and nonharmonic (fractal) components. The harmonic component was used to calculate the contribution of high (> 0.15 Hz)-frequency components to total HRV power (%HF). From the fractal component, the contribution of the fractal component to total HRV power (%fractal), the spectral exponent beta, and Hurst scaling exponent (H) were calculated. For both normal subjects and cardiac patients, atropine resulted in significantly (P < 0.05) less mean HRV and significantly (P < 0.05) greater beta compared with control, whereas mean values for %fractal were as high as 70% and were not significantly (P > 0.05) different between atropine and control. The mean value of H with atropine was significantly (P < 0.05) greater than that for control. Directional changes in %HF and beta were consistent with only one exception for a patient who had the smallest change in log %HF by atropine. The normally irregular fractal pattern of resting HRV was decreased by atropine as shown by the decrease in %HF and the increase in beta.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7485601 TI - Functional significance and control of release of pulmonary surfactant in the lizard lung. AB - The amount of pulmonary surfactant in the lungs of the bearded dragon (Pogona vitticeps) increases with increasing body temperature. This increase coincides with a decrease in lung compliance. The relationship between surfactant and lung compliance and the principal stimuli for surfactant release and composition (temperature, ventilatory pattern, and autonomic neurotransmitters) were investigated. We chose to investigate ventilatory pattern (which causes mechanical deformation of the type II cells) and adrenergic agents, because they are the major stimuli for surfactant release in mammals. To examine the effects of body temperature and ventilatory pattern, isolated lungs were ventilated at either 18 or 37 degrees C at different ventilatory regimens. An isolated perfused lung preparation at 27 degrees C was used to analyze the effects of autonomic neurotransmitters. Ventilatory pattern did not affect surfactant release, composition, or lung compliance at either 18 or 37 degrees C. An increase in temperature increased phospholipid reuptake and disproportionately increased cholesterol degradation/uptake. Epinephrine and acetylcholine stimulated phospholipid but not cholesterol release. Removal of surfactant caused a decrease in compliance, regardless of the experimental temperature. Temperature appears to be the principal determinant of lung compliance in the bearded dragon, acting directly to increase the tone of the smooth muscle. Increasing the ambient temperature may result in greater surfactant turnover by increasing cholesterol reuptake/degradation directly and by increasing circulating epinephrine, thereby indirectly increasing phospholipid secretion. We suggest that changing ventilatory pattern may be inadequate as a mechanism for maintaining surfactant homeostasis, given the discontinuous, highly variable reptilian breathing pattern. PMID- 7485602 TI - EGTA-induced disruption of epithelial cell tight junctions in the lactating caprine mammary gland. AB - The suitability of the Ca2+ chelator ethylene glycol-bis(beta-aminoethyl ether) N,N,N',N'-tetraacetic acid (EGTA) to induce disruption of mammary tight junctions (TJ) and its effect on milk secretion were investigated in six goats. EGTA was administered via the teat of one gland as an isosmotic (300 mosmol/l) K-EGTA solution (68 mM EGTA), whereas the control gland received an isosmotic sucrose solution. Lactose, Na, K, and Cl in milk, blood lactose, and the presence of Evans blue (EB) in mammary lymph were used as indicators of TJ disruption. EGTA caused transient (approximately 60 h) changes (P < 0.05) in the concentration of lactose, K, Na, and Cl in milk, consistent with loss of TJ integrity. This was confirmed by a rapid (< 1 h) increase (P < 0.05) in blood lactose levels. Moreover, EB appeared in lymph < 1 h after EGTA+EB treatment. Milk secretion declined unilaterally by 15% (P < 0.05) after EGTA and did not return to baseline until approximately 60 h after EGTA. EGTA caused a unilateral, temporary (first 7 h) increase in mammary blood flow. This study shows that a rapid temporary disruption of mammary TJ can be successfully induced in vivo and that such disruption compromises milk secretion. PMID- 7485603 TI - Castration attenuates prolactin response but potentiates ACTH response to conditioned stress in the rat. AB - The purpose of this study was to examine the effect of circulating androgens on neuroendocrine, autonomic, and behavioral responses to stress. The effects of conditioned stress were studied in male Sprague-Dawley rats that were intact, gonadectomized, or gonadectomized and treated with dihydrotestosterone (DHT). Intact animals received sham surgeries. Animals were stressed 3 wk after surgery. The adrenocorticotropic hormone (ACTH) response to conditioned stress was significantly potentiated (P < 0.01) in gonadectomized males compared with sham operated and gonadectomized DHT-treated animals. In stressed rats, plasma corticosterone levels were significantly higher (P < 0.05) in gonadectomized animals compared with DHT-treated castrates. The prolactin response to stress was decreased (P < 0.01) in gonadectomized males compared with sham-operated and gonadectomized DHT-treated rats. The stress-induced increases in plasma renin activity and concentration were not altered in gonadectomized or in gonadectomized DHT-treated animals. Nonstressed DHT-treated castrates exhibited more "fearlike" behavior compared with nonstressed sham-operated and gonadectomized animals. However, conditioned stress produced the same behavioral effects in all treatment groups. The results demonstrate that the ACTH/corticosterone, prolactin, and behavioral responses to a psychological stressor are differentially regulated by circulating androgens. PMID- 7485604 TI - Spinal angiotensin II influences reflex cardiovascular responses to muscle contraction. AB - We tested the hypothesis that inhibition of angiotensin II (ANG II) AT1 receptors in the thoracic spinal cord attenuates the reflex cardiovascular response to electrically induced hindlimb static contraction (exercise pressor reflex). Consequently, in alpha-chloralose-anesthetized cats, contraction-induced increases in mean arterial blood pressure, maximal rate of rise in left ventricular pressure (dP/dt), and heart rate were compared before and after intrathecal injection of the AT1 receptor antagonist losartan (100 or 1,000 micrograms; n = 7). Losartan significantly diminished increases in blood pressure and maximal dP/dt provoked by static contraction by 33 +/- 5 and 31 +/- 6%, respectively. Conversely, these contraction-induced responses were unaffected by similar injection of ANG II into the lumbosacral spinal cord (n = 5). Moreover, intravenous injection of 100 micrograms losartan did not affect the cardiovascular response to contraction. Our data suggest that ANG II has a excitatory effect on the efferent arm of the exercise pressor reflex, which may be due to a facilitatory action on sympathetic nerve activity. PMID- 7485605 TI - Impaired gut lipid absorptive capacity after trauma-hemorrhage and resuscitation. AB - Although the barrier function of the intestinal mucosa is impaired after hemorrhage, it remains unclear whether this is associated with a deficit in mucosal function. The aim of this study, therefore, was to determine whether trauma-hemorrhage affects the in vivo lipid absorptive capacity of the gut and, if so, to characterize the uptake process of free fatty acids in isolated enterocytes. To study this, rats were anesthetized, a laparotomy was performed (i.e., trauma was induced), and various blood vessels were cannulated. For in vivo lipid absorption, the main intestinal lymph vessel was cannulated and a jejunostomy feeding tube was inserted. The animals were bled to and maintained at a mean arterial pressure of 40 mmHg until 40% of shed blood volume was returned in the form of Ringer lactate. They were then resuscitated with four times the volume of maximal bleed out with Ringer lactate. The in vivo and in vitro lipid absorptive capacities were assessed by measuring lymph triglyceride output after a fat load and by determining the linoleic acid uptake rates on isolated enterocytes, respectively. The results show that the in vivo lipid absorption capacity of the gut is severely depressed after trauma-hemorrhage and resuscitation. Similarly, in enterocytes isolated from hemorrhaged rats, fatty acid uptake capacity, as reflected by the decreased maximal uptake rates, was significantly reduced: 1.2 +/- 0.2 and 2.6 +/- 0.6 nmol.min-1 x 10(6) cells-1 for hemorrhaged and sham, respectively. Thus gut lipid absorptive function is depressed after trauma-hemorrhage and resuscitation, which is at least partially due to the depressed uptake mechanism of the enterocyte. PMID- 7485606 TI - Body temperature modification of osmotically induced vasopressin secretion and thirst in humans. AB - We examined the effect of increased body core temperature (Tes) on the plasma arginine vasopressin concentration ([AVP]p) and thirst responses to increased plasma osmolality (Posm) induced by 3% NaCl infusion for 120 min in seven healthy humans. Tes was increased by immersion of the lower legs in 41 degrees C water in a 28 degrees C room (passive heating; HT). Immersion of the lower legs in 34.5 degrees C water on a separate day served as the control (thermoneutral; NT). The 120-min hypertonic saline infusion was initiated 30 min after the onset of leg immersion and was followed by a 30-min rehydration period. Tes in HT increased by 0.21 +/- 0.04 degree C before infusion and by 0.86 +/- 0.08 degree C at the end of infusion. The change in Tes in NT before and after the infusion was negligible. Posm was increased by 15.0 +/- 1.0 mosmol/kgH2O by infusion in both NT and HT. [AVP]p increased by 3.48 +/- 0.72 pg/ml in NT and by 7.59 +/- 1.02 pg/ml in HT. Thus the increase in [AVP]p at a given increase in Posm was markedly higher in HT than in NT. The plasma renin activity response to hypertonic saline infusion in both conditions was similar. Subjective thirst rating and cumulative water intake during rehydration were higher in HT than in NT.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7485607 TI - Effects of elevated fetal cortisol concentrations on the volume, secretion, and reabsorption of lung liquid. AB - We have examined the role of cortisol in the gestational age-related increase in the ability of epinephrine to inhibit the secretion and induce the reabsorption of fetal lung liquid. Chronically catheterized fetal sheep were infused with either saline (n = 6) or increasing doses of cortisol (1.5-3.5 mg/day; n = 6) between 120 and 130 days of gestation (term approximately 145 days). Lung liquid volumes and secretion rates were measured at 120 days (before infusion) and at 125 days, and then at 130 days we tested the ability of epinephrine to inhibit lung liquid secretion and induce liquid reabsorption. Cortisol infusions increased fetal plasma cortisol and 3,5,3'-triiodothyronine (T3) concentrations to levels observed just before labor and significantly increased the age-related increase in fetal lung liquid volume and secretion rate. At 130 days, epinephrine caused a significantly greater rate of lung liquid reabsorption in cortisol infused fetuses (10.3 +/- 2.3 ml/h) than in saline-infused fetuses (1.5 +/- 1.6 ml/h). We conclude that a premature elevation in circulating fetal cortisol concentrations, probably in conjunction with elevated T3 concentrations, prematurely increases the epinephrine-induced reabsorption of fetal lung liquid. It is likely, therefore, that the preparturient increase of fetal cortisol concentrations plays an important role in the clearance of lung liquid at birth. PMID- 7485609 TI - Dopamine D1 or D2 antagonists enhance inhibition of consummatory ingestive behavior by CCK-8. AB - A dopamine D1 (SKF-38393, 1 mg)- or D2 (LY-171555, 0.1 mg)-receptor agonist inhibited intake of an intraorally infused solution of sucrose by male rats, a test of consummatory ingestive behavior. Treatment with a D1 (SCH-39166, 0.1 mg) or D2 (raclopride, 0.6 mg) antagonist reversed inhibition by the respective agonist but enhanced the inhibitory effect of cholecystokinin octapeptide (CCK-8; 1.8 micrograms). It was not possible to demonstrate specific effects of D1 and D2 agonists on intake of pellets, a test that does not discriminate consummatory ingestive behavior from appetitive ingestive behavior, i.e., behavior used to obtain food. The results demonstrate specific involvement of dopamine D1 and D2 receptors in inhibition of consummatory ingestive behavior. PMID- 7485608 TI - Effect of oral protein loading on renal hemodynamics in human pregnancy. AB - Glomerular filtration rate (GFR) and ERPF increase approximately 50% in human pregnancy. To determine if pregnant women have additional "renal reserve," inulin and p-aminohippurate clearances (Cin, CPAH) were measured in maximally hydrated women before and after a 300-g steak meal, once during late gestation, and again > or = 3 mo postpartum. Protein loading increased Cin [106 +/- 5 (SE) to 119 +/- 4 ml/min, P < 0.003], but not CPAH (587 +/- 35 to 624 +/- 32 ml/min, NS) in the nonpregnant state, but neither clearance was altered during gestation (Cin: 156 +/- 7 to 160 +/- 9.6 ml/min, NS; CPAH: 831 +/- 36 to 899 +/- 37 ml/min, NS). A natriuresis occurred only postpartum (+142 mu eq/min, P < 0.02), which could be explained by the increased GFR alone, since indexes of filtrate delivery and reabsorption (V/GFR, CH2O/GFR, CH2O/V) and fractional sodium excretion changed little. Dopamine excretion, uninfluenced by protein, did not correlate with increments in GFR. A carbohydrate meal (time controls) had no effect on the above described parameters. We make the following conclusions. If protein and pregnancy achieve hyperfiltration by similar mechanisms, these pathways appear "exhausted" in gestation. Also, oral protein loading does not measure maximal renal reserve, since basal GFR in late gestation was substantially greater than that measured after protein feeding in nonpregnant subjects. PMID- 7485610 TI - Corpus spongiosum penis pressure and perineal muscle activity during reflexive erections in the rat. AB - This experiment represents the first simultaneous analysis of corpus spongiosum of the penis (CSP) pressure and perineal muscle activity during reflex-induced penile erections in the awake rat and provides new data on the coordination of muscular and vascular events in erection physiology. Nine male Sprague-Dawley rats were implanted with pressure transducers under pentobarbital anesthesia (60 mg/kg) for use in a new technique of chronic erection recording involving simultaneous CSP pressure monitoring and electromyography (EMG) of the ischiocavernosus (IC) and bulbospongiosus (BS) muscles. The association of these physiological measurements with visually scored ex copula erectile events was investigated. Reflexive erectile events were systematically associated with an increase in baseline CSP pressure. Glans erections were associated with dramatic suprasystolic CSP pressure peaks concurrent with BS muscle bursts. Indeed, measures of CSP pressure and BS EMG activity were found to vary significantly with the intensity of glans erection. Flips of the penile body, however, involved small CSP pressure increases and IC bursts. These data demonstrate for the first time that both CSP pressures and perineal muscle activity vary significantly with the intensity of penile erection. PMID- 7485612 TI - Effects of pulmonary denervation on renal sympathetic and heart rate responses to hypoxia. AB - We hypothesized that the renal sympathetic nerve activity (RSNA) response to hypoxia is attenuated because of stimulation of pulmonary receptors by the increase in ventilation. RSNA was measured during 20 min of severe hypoxia (8% O2) in conscious New Zealand White rabbits with intact lung innervation and in rabbits with surgical denervation of the lungs (LDX). LDX decreased resting breathing frequency but had no effect on resting mean arterial pressure (MAP), heart rate (HR), or RSNA. In intact rabbits, 4 min of hypoxia resulted in elevated RSNA (from 14 +/- 2 to 29 +/- 3% of smoke-elicited maximum), bradycardia (delta-65 +/- 12 beats/min), and no change in MAP (delta 2 +/- 2 mmHg). Bradycardia diminished with time, but elevated RSNA was maintained throughout the 20-min exposure. LDX enhanced the initial bradycardia (delta-113 +/- 11 beats/min, P < 0.01) but had no effect on the RSNA response (35 +/- 2% of maximum) to hypoxia. LDX did not alter steady-state responses of HR or RSNA, but MAP declined over time (-11 +/- 2 mmHg). These results suggest that in conscious rabbits pulmonary receptors have a minor influence on control of sympathetic activity to viscera during severe hypoxemia. PMID- 7485613 TI - Enhanced gustatory neural responses to sugars in the diabetic db/db mouse. AB - Sweet taste sensitivity in a genetic model of diabetes, the db/db mouse, in which a single major gene defect leads to the expression of diabetes and obesity, was studied by examining chorda tympani nerve responses to various taste stimuli, including sugars. The chorda tympani responses to four sugars, sucrose, fructose, glucose, and maltose, in adult db/db mice showed greater relative magnitudes and lower thresholds than those in adult lean mice, but responses to other basic taste stimuli, such as NaCl, HCl, and quinine HCl, were not different in the two groups. Behavioral experiments using a two-bottle preference test demonstrated that taste preference scores for the four sugars at suprathreshold concentrations, except 1.0 M, were higher in db/db than in control mice. Infant mice of 7-9 days of age possessing the genotype db/db also exhibited greater neural responses and lower thresholds for sugars than infant control mice, whereas streptozotocin-induced adult diabetic mice possessing the genotype +/+ did not exhibit larger sugar responses. These findings suggest that the enhanced sugar sensitivities observed in db/db mice are probably determined by a single major gene, db. The db gene may act on a common factor(s) involved in the stimulus-secretion coupling in the pancreatic B cell and the taste cell of db/db mice. PMID- 7485611 TI - Blockade of beta-adrenoceptor in control of blood pressure in fowl. AB - Several avian species show elevated blood pressure (BP) and spontaneous atherogenesis in the aorta and other large arteries. The BP appears to be influenced by age, sex (higher in males), environment, and diet in some species. We reported previously that mean aortic pressure and heart rate, but not plasma renin activity (PRA), of conscious female domestic fowl were markedly reduced by propranolol. In the present study, we aimed to determine in conscious roosters whether 1) hypotension evoked by atenolol or practolol, which selectively inhibit cardiac beta-receptors in mammals, is more potent than that evoked by propranolol, and 2) the renin-angiotensin (ANG) system and/or catecholamines are involved in beta-adrenoceptor antagonist-induced hypotension. Mean arterial pressure (171.2 +/- 3.5 mmHg) and heart rate (281 +/- 4 beats/min) of chronically cannulated roosters (n = 38) were markedly reduced by acute infusion or repeated injections (14 days) of propranolol, atenolol, or practolol, but not by SQ-14,225 (ANG-converting enzyme inhibitor) or [Sar1, Thr8]ANG II (nonselective ANG receptor antagonist). None of the beta-adrenoceptor blockers, however, showed cardioselectivity. The resting PRA of conscious roosters (1.27 +/- 0.09 ng.ml-1.h 1, n = 38) was low and did not change significantly after chronic or acute treatment with beta-adrenoceptor blockers except for a slight decrease induced by practolol. PRA increased after SQ-14,225. The plasma levels (pg/ml) of norepinephrine (701.9 +/- 76.0), epinephrine (337.2 +/- 57.1), and dopamine (299.1 +/- 39.0) of conscious roosters were further increased by propranolol. Practolol also increased dopamine significantly.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7485614 TI - Cerebral lactate metabolism in near-term fetal sheep. AB - The present study was designed to see if lactate can cross the blood-brain barrier of the near-term fetal sheep and replace glucose as an oxidative substrate during normoglycemia and acute insulin-induced hypoglycemia. Cerebral uptake of glucose, oxygen, lactate, and [14C]lactate as well as cerebral production of 14CO2 were measured under three conditions: 1) normoglycemia normolactemia, 2) acute hypoglycemia-normolactemia, and 3) hypoglycemia-steady state hyperlactemia. Although uptake of tracer [14C]lactate was consistent, there was no net uptake of unlabeled lactate during either normoglycemia or hypoglycemia. When arterial lactate concentration was raised from 2.2 +/- 0.5 to 3.3 +/- 0.4 (SE) mM by sodium lactate infusion, however, lactate was taken up. Comparison of cerebral [14C]lactate uptake with 14CO2 production indicated that the principal metabolic fate of lactate is oxidation. At increased concentrations, exogenous lactate accounted for approximately 7% of cerebral oxygen consumption. This study demonstrates that lactate crosses the blood-brain barrier of the near-term fetal sheep, is oxidized, and at elevated concentrations can partially replace glucose as an oxidative substrate during acute hypoglycemia. PMID- 7485616 TI - Examining Mnemosyne: an end to metaphors? PMID- 7485615 TI - Chronic lung lymph fistula that only drains the left lung. AB - We developed a chronic lung fistula that drains only the left lung, allowing for evaluation of injury in a single lung. To remove lymph drainage from the right lung into the caudal mediastinal lymph node, the right lower pulmonary ligament was severed. Pneumatic occluders were placed on the left pulmonary arteries and veins. To ensure that lymph drained from only the left lung, we increased the right pulmonary arterial pressure (RPAP) from 21.2 +/- 0.5 to 36.5 +/- 0.6 mmHg. The left pulmonary arterial pressure (LPAP) was kept at wedge pressure level for 1 h by inflating pneumatic occluders. Lymph flow from the left lung fistula was stable during this occlusion. Six hours after recovery was increased the LPAP from a baseline level of 19.1 +/- 1.0 to 36.4 +/- 0.9 mmHg and the RPAP from 21.2 +/- 0.5 to 38.0 +/- 0.8 mmHg for 2 h by inflating the pneumatic occluders on the left and right pulmonary veins. Lymph flow increased from 5.3 +/- 1.0 to 28.0 +/- 2.9 ml/h. Reflection coefficient was calculated at 0.80 +/- 0.02. PMID- 7485617 TI - Images in neuroscience. Neuroimaging, II. Regional cerebral blood flow with H2(15)O. PMID- 7485618 TI - The social construction of the human brain. AB - OBJECTIVE: The purpose of this article is to review the development of concepts about the contribution of nature and nurture to brain structure and mental function, and to derive the implications of these changing concepts for clinical practice. METHOD: The literature of the past five decades, as refracted by the author's personal experience in academic psychiatry during that interval, is reviewed. RESULTS: Psychiatric theory has swung through mighty arcs in recent years but has begun to re-equilibrate. Fifty years ago, psychoanalysis dominated the academic scene; for the past two decades, reductionist biological determinism has held the fort. Neither position is tenable. To subscribe to either is possible only by ignoring conflicting evidence. Worse, it means short-changing patients, whose disorders do not come neatly packaged into "organic" and "functional" compartments. Development is neither predestined in the genome nor completely malleable to shaping by the environment. Children inherit, along with their parents' genes, their parents, their peers, and the communities they inhabit. CONCLUSIONS: Contemporary psychiatric research conclusively demonstrates that mind/brain responds to biological and social vectors and is jointly constructed by both. Major brain pathways are specified in the genome; detailed connections are fashioned by, and consequently reflect, socially mediated experience in the world. Just at the time when integration at the level of theory is coming into sight, comprehensive patient care is endangered by for-profit corporate managed care, which is transforming medical visits into commodities on a production line. Physicians and patients must join in a coalition to protect quality, ensure access, and build continuity into all of medical care. PMID- 7485619 TI - Remembering the past: two facets of episodic memory explored with positron emission tomography. AB - OBJECTIVE: This study used positron emission tomography to examine two kinds of personal memory that are used in psychiatric evaluation: focused episodic memory (recall of past experience, employed in "taking a history") and random episodic memory (uncensored thinking about experience, examined during analytic therapy using free association). For comparison, a third memory task was used to tap impersonal memory that represents general information about the world ("semantic memory"). METHOD: Thirteen subjects were studied using the [15O]H2O method to obtain quantitative measurements of cerebral blood flow. The three conditions were subtracted and their relative relationships examined. RESULTS: The random episodic condition produced activations in widely distributed association cortex (right and left frontal, parietal, angular/supramarginal, and posterior inferior temporal regions). Focused episodic memory engaged a network that included the medial inferior frontal regions, precuneus/retrosplenial cingulate, anterior cingulate, thalamus, and cerebellum. The use of medial frontal regions and the precuneus/retrosplenial cingulate was common to both focused and random episodic memory. The major difference between semantic and episodic memory was activation of Broca's area and the left frontal operculum by semantic memory. CONCLUSIONS: These results indicate that free-ranging mental activity (random episodic memory) produces large activations in association cortex and may reflect both active retrieval of past experiences and planning of future experiences. Focused episodic memory shares some components of this circuit (inferior frontal and precuneus), which may reflect the time-linked components of both aspects of episodic memory, and which permit human beings to experience personal identity, consciousness, and self-awareness. PMID- 7485620 TI - Psychopathology from adolescence into young adulthood: an 8-year follow-up study. AB - OBJECTIVE: This study investigated the stability of behavioral and emotional problems from adolescence into young adulthood. METHOD: Subjects from the general population (N = 459), aged 13-16 years, were evaluated initially with the Child Behavior Checklist (completed by parents) and 8 years later with the Young Adult Self-Report. The scoring format and factor structure of the two assessment instruments are similar; syndromes constructed from the two instruments are based on parents', teachers', and self-report information derived from large clinical samples. Signs of maladjustment also were assessed at follow-up through interviews. RESULTS: Of the individuals with total problem scores in the deviant range on the Child Behavior Checklist, 27.3% had total problem scores in the deviant range on the Young Adult Self-Report at follow-up. The probability of having a total problem score in the deviant range at follow-up was raised 7.4 fold by having deviant-range scores on the Child Behavior Checklist somatic complaints and anxious/depressed syndromes (simultaneously) at the initial assessment. Referral to mental health services was predicted by deviant-range scores on the anxious/depressed syndrome, while suicide attempts were predicted by deviance on the withdrawn syndrome. CONCLUSIONS: Adolescent problems tended to persist into young adulthood to a moderate degree. High rates of withdrawal from social contacts, anxiety or depression, somatic complaints without known medical origin, social problems, attention problems, delinquent behavior, and aggressive behavior during adolescence were risk factors for specific types of psychopathology and maladjustment at 8-year follow-up. The presence of psychopathology in adolescence should not be regarded as normative. PMID- 7485621 TI - High autonomic arousal and electrodermal orienting at age 15 years as protective factors against criminal behavior at age 29 years. AB - OBJECTIVE: Nothing is known about biological factors that protect a predisposed individual from becoming criminal. This 14-year prospective study tested the hypothesis that antisocial adolescents who desist from crime by age 29 have greater physiological arousal and orienting than antisocial adolescents who become adult criminals. METHOD: Physiological arousal and orienting were measured in 101 unselected 15-year-old male schoolchildren. Of these, 17 antisocial adolescents who desisted from adult crime (desistors) were matched on adolescent antisocial behavior and demographic variables with 17 antisocial adolescents who became criminal by age 29 (criminals), and 17 nonantisocial, noncriminal normal subjects. RESULTS: Desistors had significantly higher electrodermal and cardiovascular arousal and higher electrodermal orienting than the criminal group. CONCLUSIONS: This is the first study to report biological factors that protect against the development of criminal behavior. The findings suggest that individuals predisposed to adult crime by virtue of showing antisocial behavior in adolescence may be protected from committing crime by high levels of autonomic arousal and orienting. PMID- 7485622 TI - Clinical assessment versus research methods in the assessment of suicidal behavior. AB - OBJECTIVE: This study examined how accurately routine inpatient clinical assessments documented a history of overt suicidal behavior in inpatients with a diagnosis of major depressive episode. Secondary questions involved the exploration of possible factors influencing the quality of routine clinical documentation of suicidal behavior, such as lethality of attempts, axis II comorbidity, and presence of recent suicidal behavior. METHOD: Hospital records for 50 patients, known to have a history of suicidal behavior on the basis of research ratings, were reviewed to assess reporting of the number of lifetime suicide attempts, suicidal ideation and planning behavior, most medically lethal suicide attempt, and family history of suicidal behavior. These measures of suicidal behavior were compared with a comprehensive research assessment, completed concurrently and independently. RESULTS: At admission clinicians failed to document a history of suicidal behavior in 12 of 50 patients identified by research assessment as depressed and as having attempted suicide. Fewer total suicide attempts were clinically reported than in research data. Documentation of suicidal behavior was least accurate in the physician discharge summary and was most accurate on hospital intake assessment, which employed a semistructured format for recording clinical information including suicidal behavior. CONCLUSIONS: A significant degree of past suicidal behavior is not recorded during routine clinical assessment, and the use of semistructured screening instruments may improve documentation and detection of lifetime suicidal behavior. The physician discharge summary must accurately document suicidal behavior, since it best identified a high-risk population for out-patient clinicians responsible for follow-up. PMID- 7485623 TI - Validity of a diagnosis of lifetime major depression obtained by personal interview versus family history. AB - OBJECTIVE: Diagnoses obtained by family history agree only modestly with those obtained through personal interview. If personal interview diagnoses are the "gold standard," these findings suggest that family history diagnoses have low validity. Here the authors take another perspective--to evaluate family history versus personal interview diagnoses of lifetime major depression by three independent validators. METHOD: In a large sample of female-female twin pairs and their parents (903 families) ascertained from a population-based twin register, all subjects were personally interviewed by using a modified Structured Clinical Interview for DMS-III-R. Family history diagnoses based on the Family History Research Diagnostic Criteria were obtained by questioning each participant about his or her relatives. By means of multiple regression, the powers of the personal interview and family history methods were compared to predict 1) future episodes of major depression in the twins, 2) neuroticism, and 3) familial aggregation of major depression. RESULTS: Agreement between diagnoses obtained by personal interview and family history was modest. After the presence or absence of a personal interview diagnosis of major depression was controlled for, a family history diagnosis of major depression significantly predicted future episodes of major depression, neuroticism (in five of six analyses), and familial aggregation of major depression (in four of six analyses). CONCLUSIONS: Although agreeing relatively poorly, diagnoses of lifetime major depression obtained by personal interview and family history both contained useful information about future episodes, personality, and familial liability to illness. A multimethod approach to assessment of psychiatric illness may maximize the validity of psychiatric diagnoses. PMID- 7485624 TI - Who comes to a voluntary depression screening program? AB - OBJECTIVE: The purpose of this study was to describe who comes to a voluntary depression screening program by analyzing findings from the 1992 National Depression Screening Day. METHOD: Survey results from 5,367 adult volunteers at 345 facilities were analyzed. The authors examined the prevalence of depression detected at the screening test and sociodemographic characteristics and treatment history of the respondents. They also estimated the percentage of these individuals who would actually have a diagnosis of major depression (positive predictive value). The main assessment measure was the Zung Self-Rating Depression Scale. RESULTS: Of all participants, 76.6% (N = 4,109) had at least minimal depressive symptoms (score of at least 50 on the Zung depression scale), 53.3% (N = 2,859) had at least moderate symptoms (score of at least 60), and 22.6% (N = 1,214) had severe symptoms (score of at least 70). Few of the depressed respondents were currently in treatment for a mental health problem. The positive predictive value of a screening test diagnosis of depression was between 92.5% and 95.5% when a cutoff score of 60 was used to indicate depression and between 88.7% and 92.3% when a cutoff score of 50 was used. CONCLUSIONS: Voluntary screening tests, as exemplified by results from the National Depression Screening Day, provide a good opportunity for identifying previously unidentified and untreated individuals with depression. PMID- 7485625 TI - Body weight, dieting, and eating disorder symptoms among college students, 1982 to 1992. AB - OBJECTIVE: The authors sought to examine changes in prevalence of dieting behavior and eating disorder symptoms from 1982 to 1992. METHOD: In 1982, 625 women and 276 men participated in a study examining body weight, eating habits, dieting tendencies, and eating disorder symptoms. Ten years later 564 women and 235 men at the same college completed a nearly identical survey. Similar random sampling methods were used for both studies. All respondents were classified into one of five groups (nondieter, dieter, problem dieter, subclinical eating disorder, or eating disorder according to DSM-III-R criteria). RESULTS: On almost all measures there were significant reductions of problematic eating behaviors and disordered attitudes about body, weight, and shape from 1982 to 1992. The estimated prevalence of bulimia nervosa dropped from 7.2% to 5.1% for women and from 1.1% to 0.4% for men. Binge eating, vomiting, diuretic use, and diet pill use also declined for women during this period. Significantly fewer women and men reported chronic dieting in 1992 than in 1982, and there was evidence of improved body image for both sexes. Subjects in 1992 also reported healthier eating habits in terms of dietary intake and meal regularity. Finally, women in 1992 were more likely to be overweight and were, on average, five pounds heavier than their 1982 counterparts. CONCLUSIONS: The prevalence of problematic eating behaviors and eating disorder symptoms appears to be abating. However, they remain a significant problem that affects a substantial segment of this population. PMID- 7485626 TI - Persistent perfectionism, symmetry, and exactness after long-term recovery from anorexia nervosa. AB - OBJECTIVE: It is well recognized that patients with anorexia nervosa have perfectionistic and obsessive behaviors. This study investigated whether such behaviors persist after recovery. METHOD: Twenty subjects who had recovered from anorexia nervosa were recruited for the study. They had been at normal weight and their menses had been regular for more than a year. These subjects were compared with 16 healthy women with the use of the Eating Disorder Inventory, the Frost Multidimensional Perfectionism Scale, and the Yale-Brown Obsessive Compulsive Scale. RESULTS: The recovered anorexic patients had significantly higher scores than the comparison women on the measures of perfectionism on the Eating Disorder Inventory and on overall perfectionism on the Frost scale. Moreover, the recovered patients had higher scores on the Yale-Brown scale, with target symptoms suggesting that many had specific concerns with symmetry and exactness. CONCLUSIONS: Certain characteristics of anorexia nervosa, such as a need for order and precision, persist after good outcome and recovery, raising the question of whether these behaviors are traits that contribute to the pathogenesis of this illness. PMID- 7485627 TI - Relapse and impairment in bipolar disorder. AB - OBJECTIVE: The purpose of this study was to evaluate the outcome of bipolar disorder in the context of maintenance pharmacotherapy. METHOD: Eighty-two bipolar outpatients were followed prospectively for a mean of 4.3 years (minimum of 2 years); symptom rating and psychosocial outcome scales were used, and pharmacotherapy was rated on a 5-point scale. RESULTS: Despite continual maintenance treatment, survival analysis indicated a 5-year risk of relapse into mania or depression of 73%. Of those who relapsed, two-thirds had multiple relapses. Relapse could not be attributed to inadequate medication. Even for those who did not relapse, considerable affective morbidity was observed. A measure of cumulative affective morbidity appeared to be a more sensitive correlate of psychosocial functioning than was the number of relapses. Poor psychosocial outcome paralleled poor syndromal course. Poor psychosocial functioning, especially occupational disruption, predicted a shorter time to relapse. Depressions were most strongly related to social and family dysfunction. CONCLUSIONS: Even aggressive pharmacological maintenance treatment does not prevent relatively poor outcome in a significant number of bipolar patients. PMID- 7485628 TI - Postpartum prophylaxis for women with bipolar disorder. AB - OBJECTIVE: The postpartum period has typically been considered a time of heightened vulnerability for development of affective disorders, and women with bipolar disorder have consistently demonstrated vulnerability to puerperal worsening of mood. This retrospective study examined the extent to which mood stabilizing agents provide prophylactic benefit to bipolar women during the postpartum period. METHOD: The clinical course of 27 women meeting the DSM-III-R criteria for bipolar disorder was followed during pregnancy and the postpartum period. Information regarding severity of illness (as measured by number of episodes of mania, depression, or both) was obtained, in addition to data on pharmacotherapy (if any) received before, during, and after pregnancy. The extent to which the prophylactic use of antimanic agents minimized the risk of relapse was explored. RESULTS: Only one of the 14 patients taking prophylactic agents during the acute puerperium relapsed within the first 3 months postpartum, while eight of the 13 who did not receive antimanic drugs showed evidence of recurrent affective instability during those 3 months. A survival analysis indicated that the women receiving prophylactic treatment with mood stabilizers maintained well being significantly longer than the women who did not receive such treatment. CONCLUSIONS: Women with bipolar disorder appear to benefit from puerperal prophylaxis with mood stabilizers. Consistent with results of earlier studies, postpartum prophylaxis was associated with lower rates of relapse into affective disorders. The findings have implications for the early identification and treatment of subgroups of women at particular risk for puerperal illness. PMID- 7485629 TI - Comorbid Tourette's disorder and bipolar disorder: an etiologic perspective. AB - OBJECTIVE: Using an epidemiologic approach, the authors attempt to elucidate relationships between Tourette's disorder and bipolar disorder. METHOD: Of 205 patients with Tourette's disorder in the North Dakota Longitudinal Tourette Syndrome Surveillance Project, 15 had comorbid bipolar disorder. A subset of the patients with Tourette's disorder had been included in earlier population-based prevalence studies of Tourette's disorder in children, adolescents, and adults. Minimal risk ratios were calculated for the patients with Tourette's disorder plus bipolar disorder by age group (children/adolescents and adults). This information was used to estimate genetic risk indicators for comorbid Tourette's disorder and bipolar disorder. RESULTS: The estimated risk of developing bipolar disorder among the study group of children, adolescents, and adults with Tourette's disorder was more than four times higher than the level expected by chance, but this finding did not reach statistical significance. It was indicative of trends, however. CONCLUSIONS: Comorbidity between Tourette's disorder and bipolar disorder does not appear to be due to chance co-occurrence of the two disorders. Although a genetic mechanism may play a causal role, in the absence of family studies an explanatory model involving the concept of canalization of basal-ganglia-mediated dysfunctions is offered. In such a construct, Tourette's disorder would be a likely accompaniment to other conditions, including bipolar disorder, whose pathogenic determinants might channel through neural pathways involving the basal ganglia. The presence of significant developmental disabilities may further enhance factors culminating in comorbid Tourette's disorder and bipolar disorder. PMID- 7485630 TI - Psychoactive substance use disorders in adults with attention deficit hyperactivity disorder (ADHD): effects of ADHD and psychiatric comorbidity. AB - OBJECTIVE: The authors evaluated the association between attention deficit hyperactivity disorder (ADHD) and psychoactive substance use disorders in adults with ADHD, attending to comorbidity with mood, anxiety, and antisocial disorders. It was hypothesized that psychiatric comorbidity would be a risk factor for psychoactive substance use disorders. METHOD: Findings for 120 referred adults with a clinical diagnosis of childhood-onset ADHD were compared with those for non-ADHD adult comparison subjects (N = 268). All childhood and adult diagnoses were obtained by structured psychiatric interviews for DSM-III-R. RESULTS: There was a significantly higher lifetime risk for psychoactive substance use disorders in the ADHD adults than in the comparison subjects (52% versus 27%). Although the two groups did not differ in the rate of alcohol use disorders, the ADHD adults had significantly higher rates of drug and drug plus alcohol use disorders than the comparison subjects. ADHD significantly increased the risk for substance use disorders independently of psychiatric comorbidity. Antisocial disorders significantly increased the risk for substance use disorders independently of ADHD status. Mood and anxiety disorders increased the risk for substance use disorders in both the ADHD and comparison subjects, but more demonstrably in the comparison subjects. CONCLUSIONS: Although psychiatric comorbidity increased the risk for psychoactive substance use disorders in adults with ADHD, by itself ADHD was a significant risk factor for substance use disorders. More information is needed to further delineate risk and protective factors mediating the development of substance use disorders in persons with ADHD. PMID- 7485631 TI - Sleep disturbance and its relationship to psychiatric morbidity after Hurricane Andrew. AB - OBJECTIVE: Sleep disturbance is an important dimension of posttraumatic stress disorder (PTSD), but most of the limited available data were obtained years after the original traumatic event. This study provides information on sleep disturbance and its relationship to posttraumatic morbidity from evaluations done within a year after the trauma. METHOD: Sleep and psychiatric symptoms of 54 victims (12 men and 42 women) of Hurricane Andrew who had no psychiatric illness in the 6 months before the hurricane were evaluated. A subset of hurricane victims with active psychiatric morbidity (N = 10) and nine comparison subjects who were unaffected by the hurricane were examined in a sleep laboratory. RESULTS: A broad range of sleep-related complaints were rated as being greater after the hurricane, and psychiatric morbidity (which was most commonly PTSD, followed by depression) had a significant effect on most of the subjective sleep measures. In addition, subjects with active morbidity endorsed greater frequencies of "bad dreams" and general sleep disturbances before the hurricane. Polysomnographic results for the hurricane victims revealed a greater number of arousals and entries into stage 1 sleep. REM density correlated positively with both the PTSD symptom of reexperiencing trauma and global distress. CONCLUSIONS: Subjects affected by Hurricane Andrew reported sleep disturbances, particularly those subjects with psychiatric morbidity. Tendencies to experience bad dreams and interrupted sleep before a trauma appear to mark vulnerability to posttraumatic morbidity. Results of sleep laboratory evaluations suggested brief shifts toward higher arousal levels during sleep for PTSD subjects and a relationship of REM phasic activity and symptom severity. PMID- 7485632 TI - Body dysmorphic disorder in patients with anxiety disorders and major depression: a comorbidity study. AB - OBJECTIVE: The authors evaluated the frequency of body dysmorphic disorder in patients with a primary diagnosis of anxiety disorders and major depression. METHOD: Patients with social phobia (N = 54), obsessive-compulsive disorder (N = 53), generalized anxiety disorder (N = 32), panic disorder (N = 47), and major depression (N = 42) and normal comparison subjects (N = 33) were studied. RESULTS: Body dysmorphic disorder was most common in patients with social phobia (11%) and obsessive-compulsive disorder (8%); it was less prevalent among patients with panic disorder (2%), generalized anxiety disorder (0%), and major depression (0%) and among normal subjects (0%). CONCLUSIONS: These findings suggest that body dysmorphic disorder may share etiologic elements with social phobia and obsessive-compulsive disorder. PMID- 7485633 TI - Acute tryptophan depletion and increased food intake and irritability in bulimia nervosa. AB - OBJECTIVE: Data suggest that serotonin activity is reduced in women at normal weight who have bulimia nervosa. The authors tested whether acute perturbations in serotonin activity could alter short-term eating behavior and mood. METHOD: They examined the effect of acute tryptophan depletion in 10 women with and 10 women without bulimia nervosa. RESULTS: Women with bulimia nervosa exhibited an increase in caloric intake and mood irritability after acute tryptophan depletion. CONCLUSIONS: These results indicate that women with bulimia nervosa have an exaggerated or pathological response to transient alterations in serotonin activity. PMID- 7485634 TI - Narcissism and empathy in steroid users. AB - OBJECTIVE: In an effort to begin to construct a psychological profile of anabolic steroid users, the authors compared weight lifters and bodybuilders who did or did not use anabolic steroids on an objective measure of narcissism and on clinical ratings of empathy. METHOD: The subjects were 16 weight lifters and bodybuilders who reported that they had used anabolic steroids within the past year and a comparison group of 20 weight lifters who had not used steroids. The Narcissistic Personality Inventory and clinical ratings of empathy were used to assess narcissism. RESULTS: Steroid users had significantly higher scores on dimensions of pathological narcissism and significantly lower scores on clinical ratings of empathy. CONCLUSIONS: These preliminary results document a relationship between anabolic steroid use and narcissistic personality traits. They also indicate the need for further research to determine whether narcissistic personality traits contribute to the initiation of anabolic steroid use or result from their use. PMID- 7485635 TI - Effect of previous trauma on acute plasma cortisol level following rape. AB - OBJECTIVE: The authors examined the relationships among history of previous assault, severity of rape, acute plasma cortisol level after rape, and development of rape-related post-traumatic stress disorder (PTSD). METHOD: Blood samples were drawn from 37 adult female rape victims within 51 hours after they had been raped. The subjects were assessed for history of previous assault and for the presence of PTSD 17-157 days (mean = 90 days) after the rape. RESULTS: Women with a history of previous assault had a lower mean acute cortisol level after the rape but a higher probability of subsequently developing PTSD. A significant interaction between history of previous assault and the severity of the index rape was observed: only women who had never been assaulted before had higher cortisol levels following high-severity rapes (those which included injury or multiple types of penetration) than low-severity rapes. CONCLUSIONS: The authors conclude that previous traumatization may attenuate the acute cortisol response to trauma. PMID- 7485636 TI - Does ischemia occur with panic attacks? AB - OBJECTIVE: The authors examined ambulatory ECG changes during panic attacks in patients with panic disorder. METHOD: Ten otherwise healthy women with panic disorder and complaints of chest pain during panic attacks underwent a symptom limited exercise test on a treadmill and then wore an ambulatory ECG monitor with software designed to detect and record ischemic events and used a hand-held computer for up to 6 days. RESULTS: Eight of the women had panic attacks while using the hand-held computer and the ECG monitor. No ischemic events occurred during any of the exercise tests. Twelve tachycardiac events occurred during panic attacks and 84 tachycardiac events occurred that were not associated with panic attacks. Panic attacks were associated with significantly more symptoms than were tachycardiac episodes. CONCLUSIONS: In this group of otherwise healthy women with panic disorder and chest pain, ambulatory ischemic changes were not recorded during panic attacks. PMID- 7485637 TI - Pregnancy and estrogen-induced panic. PMID- 7485638 TI - Fluvoxamine treatment of compulsivity, impulsivity, and aggression. PMID- 7485639 TI - Possible paroxetine-zolpidem interaction. PMID- 7485640 TI - Depersonalization treated with fluoxetine. PMID- 7485641 TI - Cross-tapering clozapine and risperidone. PMID- 7485642 TI - Efficacy of medium-dose clozapine for treatment-resistant schizophrenia. PMID- 7485643 TI - Inclusion of psychological torture in PTSD criterion A. PMID- 7485644 TI - On the words "mental illness". PMID- 7485645 TI - Schizophrenia: a 100-year retrospective. PMID- 7485646 TI - Schizophrenia: a 100-year retrospective. PMID- 7485647 TI - Schizophrenia: a 100-year retrospective. PMID- 7485648 TI - Schizophrenia: a 100-year retrospective. PMID- 7485649 TI - Pitfalls of SPECT studies of acute ethanol-induced changes in cerebral blood flow. PMID- 7485650 TI - Contingencies for dangerous behavior. PMID- 7485651 TI - Contingencies for dangerous behavior. PMID- 7485652 TI - Fluoxetine, buspirone, myoclonus, and dystonia. PMID- 7485653 TI - Cardiovascular morbidity and ECT. PMID- 7485654 TI - On a biological regulation of aggression. PMID- 7485655 TI - Practice guideline for the treatment of patients with substance use disorders: alcohol, cocaine, opioids. American Psychiatric Association. PMID- 7485656 TI - Out of the vortex--neonatologists' treatment decisions for newborns at risk for HIV. PMID- 7485657 TI - More on women and the prevention of HIV infection. PMID- 7485658 TI - Casual sex and HIV transmission. PMID- 7485659 TI - Annotation: needle exchange programs--do they work? PMID- 7485660 TI - Risk factors for HIV and other sexually transmitted diseases and prevention practices among US heterosexual adults: changes from 1990 to 1992. AB - OBJECTIVES: The National AIDS Behavioral Survey (1990-1992) of heterosexual adults (18-49 years) measured human immunodeficiency virus (HIV) risk factors, condom use, and HIV antibody testing, with a focus on major "high-risk" cities. METHODS: A longitudinal survey was conducted. RESULTS: There was little reduction in the overall prevalence of HIV risk factors in the national or high-risk cities cohorts over time. Despite this picture of stability, approximately 39% of the population at risk for HIV because of multiple sexual partners turns over annually. There was little change in HIV test-seeking or in consistent condom use with primary sexual partners. Although the majority of at-risk respondents used condoms sporadically or not at all (65%), a significant increase in condom use was found among those reporting multiple sexual partners in both waves, particularly among Black heterosexuals. Data from other surveys and condom sales nationally support the findings. CONCLUSIONS: There is a need for a series of surveys in this area to assess the reliability of the present findings and to monitor the general US population's response to prevention programs. PMID- 7485661 TI - HIV-1 seroconversion and risk behaviors among young men in the US army. The Seroconversion Risk Factor Study Group. AB - OBJECTIVES: This study sought to examine risk factors associated with human immunodeficiency virus type 1 (HIV-1) seroconversion among active-duty men in the US Army. METHODS: One hundred twenty-eight men with documented HIV-1 seroconversion between 1988 and 1991 were matched to control subjects on demographic variables. Risk factor information was collected for the seroconversion period. RESULTS: Forty-nine case subjects and no control subjects reported same-gender sex; this includes 34 case subjects who also reported sex with women. Seventy case and 118 control subjects reported no risk factors other than heterosexual intercourse. Among heterosexuals, excess risk was noted for men who had sex with women in risk categories defined by the Centers for Disease Control and Prevention (odds ratio = 10.0; 95% confidence interval = 1.3, 78.1). Significant trends of increasing risk for seroconversion were found with increasing numbers of female partners, nonsteady partners, and partners with whom sex occurred on the first day of acquaintance. CONCLUSIONS: In this population, the major risk factor for HIV-1 seroconversion was same-gender sex. Among heterosexuals, sex with anonymous or causal partners increased this risk. Intervention programs should emphasize the risk of indiscriminate partner selection in addition to "safe sex" practices. PMID- 7485662 TI - The treatment of non-HIV-related conditions in newborns at risk for HIV: a survey of neonatologists. AB - OBJECTIVES: The purpose of this study was to examine attitudes of neonatologists about treatment of conditions unrelated to the human immunodeficiency virus (HIV) for critically ill newborns at risk for HIV. METHODS: Questionnaires were mailed to the 1508 members of the Section on Perinatal Medicine of the American Academy of Pediatrics; 63% completed the survey (n = 951). The survey included structured questions about treatment for hypothetical cases and open-ended questions eliciting reasons for decisions. RESULTS: Differences in recommendations for treatment by both maternal and infant HIV status were substantial and statistically reliable. For example, 98% of respondents recommended life-saving cardiac surgery for a neonate with no risk for HIV, but only 93% recommended such surgery for a child of an HIV-positive mother; only 50% recommended the same surgery for a newborn known to be infected. The corresponding figures for chronic dialysis were 91%, 61%, and 26%. Most expected diminished quality of life for both infected and uninfected children of HIV-positive mothers. CONCLUSIONS: Recommendations about life-sustaining treatment for non-HIV-related conditions varied by HIV status. These data on physician attitudes raise the possibility that infants labeled as HIV positive, whether infected or not, may suffer discrimination. PMID- 7485664 TI - Reducing the risk of HIV infection among South African sex workers: socioeconomic and gender barriers. AB - OBJECTIVES: The social context within which women engaged in sex work at a popular truck stop in South Africa are placed at risk of human immunodeficiency virus (HIV) infection and the factors that influence their ability to reduce their risk were assessed. METHODS: Using qualitative and quantitative techniques, an elected sex worker from within the group collected all data. RESULTS: Given the various pressing needs for basic survival, the risk of HIV infection is viewed as one more burden imposed on these women by their lack of social, legal, and economic power. Violence, or the threat thereof, plays an important role in their disempowerment. In the few instances in which sex workers were able to insist on condom use, it resulted in a decrease in earnings, loss of clients, and physical abuse. CONCLUSIONS: Recommendations to reduce the sex workers' risk for HIV infection include negotiation and communication skills to enable them to persuade their clients to use condoms; development of strategies through which they can maximally use their group strength to facilitate unified action; and accessibility of protective methods they can use and control, such as intravaginal microbicides. PMID- 7485663 TI - Condom use among female commercial sex workers in Nevada's legal brothels. AB - OBJECTIVES: The purpose of this study was to evaluate condom use and the incidence of breakage and slippage during vaginal intercourse among female prostitutes in legal Nevada brothels, where use of condoms is required by law. METHODS: Forty-one licensed prostitutes in three brothels were enrolled in a prospective trial in August 1993. Used condoms were collected to verify reported breaks visually. Retrospective breakage and slippage rates were obtained in a standardized interview. RESULTS: Condoms were used for every act of vaginal intercourse with a brothel client during the study period, as well as in the previous year. In the prospective study phase, condoms were used in 353 acts of vaginal intercourse with clients. No condoms broke, and none fell off the penis during intercourse. Only twice (0.6%) did condoms completely fall off during withdrawal. Twelve times (3.4%) during intercourse and 15 times (4.3%) during withdrawal, condoms slipped down the penis but did not fall off. CONCLUSIONS: These findings, among the lowest breakage and slippage rates published, suggest that regular condom use may lead to condom mastery and the development of techniques to reduce the likelihood of breakage and slippage. PMID- 7485665 TI - Quality of condom use as reported by female clients of a family planning clinic. AB - OBJECTIVES: This study analyzed the prevalence and determinants of the quality of condom use after a woman's first visit to a family planning clinic. METHODS: This report presents data from 360 female family planning clients who reported using condoms as their primary method of contraception for at least 1 sexually active month during the study's follow-up period after their first clinic visit. RESULTS: Only 1% always engaged in all five use behaviors studied: using a condom for each sexual intercourse, putting the condom on before first entry, withdrawal after intercourse while there is still an erection, holding on to the condom's rim during withdrawal, and using foam. Multiple linear regression indicated that more effective condom use was reported by women who had not had an induced abortion, were not using condoms just because they were starting oral contraceptive pill regimens, had more knowledge about birth control in general, had received a nursing intervention, and had more communication with their partner. CONCLUSIONS: Increasing attention needs to be devoted to understanding determinants of the specific actions that differentiate between more and less effective contraceptive use. PMID- 7485666 TI - Reduced risk of hepatitis B and hepatitis C among injection drug users in the Tacoma syringe exchange program. AB - OBJECTIVES: This case-control study examined the association between syringe exchange use and hepatitis B and C in injection drug users. METHODS: Case patients included 28 injection drug users with acute hepatitis B and 20 with acute hepatitis C reported to the health department in a sentinel hepatitis surveillance county; control subjects were injection drug users with no markers of exposure to hepatitis B or C (n = 38 and 26, respectively) attending health department services during the same period. Data were abstracted from clinic records. RESULTS: Seventy-five percent of case patients with hepatitis B and 26% of control subjects had never used the exchange; similar proportions were found for the hepatitis C case and control groups. After adjustment for demographic characteristics and duration of injecting drugs, nonuse of the exchange was associated with a sixfold greater risk of hepatitis B (odds ratio [OR] = 5.5; 95% confidence interval [CI] = 1.5, 20.4) and a sevenfold greater risk of hepatitis C (OR = 7.3; 95% CI = 1.6, 32.8). CONCLUSIONS: The results suggest that use of the exchange led to a significant reduction in hepatitis B and hepatitis C in the county and may have also prevented a substantial proportion of human immunodeficiency virus infections in injection drug users. PMID- 7485667 TI - Risk factors for HIV-1 seroconversion among injection drug users: a case-control study. AB - OBJECTIVES: A nested case-control study was conducted in Miami, Fla, to determine risk factors associated with human immunodeficiency virus type 1 (HIV-1) seroconversion among injection drug users. METHODS: The study identified 21 incident cases of HIV-1 infection and 76 unmatched controls from two longitudinal cohorts of injection drug users. One cohort consisted of individuals who originally had been recruited from treatment centers; a second cohort was recruited from the "street." Logistic regression analyses that adjusted for age, gender, and race were performed. RESULTS: The final model determined that the primary independent risk factor that best explained the risk for seroconversion was sharing injection equipment in the year prior to conversion; a marginal risk factor was presence of sexually transmitted disease during this same period. CONCLUSIONS: Both an injection component and a sexual component play a role in seroconversion among injection drug users, although the injection component is much stronger. PMID- 7485668 TI - Sociodemographic characteristics and sexual behavior of bisexual men in France: implications for HIV prevention. The French National Survey on Sexual Behavior Group. AB - The French National Survey on Sexual Behavior was used to identify sociodemographic characteristics and sexual behavior of bisexually active men, as distinct from both homosexually and heterosexually active men. In regard to number of partners and frequency of unprotected vaginal sex, bisexuals were similar to multipartnered heterosexuals. On sociodemographic criteria, they differed from both homosexuals and heterosexuals. Bisexual men reported fewer partners than homosexuals but seemed more likely to engage in risk behavior related to the acquired immunodeficiency syndrome (AIDS), and they were less likely to have ever had a human immunodeficiency virus (HIV) test. These results justify prevention efforts specially adapted for bisexuals. PMID- 7485669 TI - Socioeconomic differences in sexually transmitted disease rates among black and white adolescents, San Francisco, 1990 to 1992. AB - This paper examines the effect of socioeconomic position on the differences in the 3-year rates (1990 to 1992) of reported cases of gonorrhea and chlamydia between Black and White adolescents, aged 12 to 20 years, residing in San Francisco. The crude relative risks for Blacks were 23.4 (95% confidence interval [CI] = 20.4, 27.8) for gonorrhea and 9.3 (95% CI = 8.3, 10.3) for chlamydia. Adjusting for poverty and occupational status, the relative risks were 28.7 (95% CI = 22.5, 36.1) for gonorrhea and 8.9 (95% CI = 7.4, 10.6) for chlamydia. This study demonstrates that factors other than poverty and occupational status account for the racial/ethnic differences in the rates of gonorrhea and chlamydia among adolescents in San Francisco. PMID- 7485670 TI - HIV seroprevalence and risk behaviors among lesbians and bisexual women in San Francisco and Berkeley, California. AB - Few data are available on human immunodeficiency virus (HIV) infection and risk behaviors among lesbians and bisexual women. A total of 498 lesbians and bisexual women was sampled from public venues in San Francisco and Berkeley, Calif, during 1993. The overall HIV seroprevalence was 1.2%. Ten percent of participants reported injecting drugs since 1978. Forty percent of the participants reported unprotected vaginal or anal sex with men during the past 3 years, including unprotected sex with gay and bisexual men and male injection drug users. The high rates of injection drug use and unsafe sexual behaviors suggest that lesbians and bisexual women frequenting public venues in San Francisco and Berkeley are at risk for HIV infection. PMID- 7485671 TI - The migration of persons with AIDS: data from 12 states, 1985 to 1992. AIDS Mortality Project Group. AB - This study measured the migration of persons with the acquired immunodeficiency virus (AIDS) between diagnosis and death using AIDS case and death reports from 12 states for 1985 to 1992. Of 49,805 persons with AIDS, 10.6% changed their place of residence, and half of these individuals who moved changed their state of residence. Migration had relatively little impact on the numbers of persons with AIDS in the largest metropolitan areas, which accounted for approximately 90% of AIDS diagnoses. Although only 3% of deaths occurred in residents of nonmetropolitan areas, the net effect of migration was a 24% increase in the number of persons with AIDS residing in such areas. PMID- 7485672 TI - Tuberculosis screening among homeless persons with AIDS living in single-room occupancy hotels. AB - Congregate facilities for homeless persons with the acquired immunodeficiency syndrome (AIDS) are often endemic for tuberculosis. We evaluated tuberculosis screening methods at single-room-occupancy hotels housing persons with AIDS. Residents were screened by cross matching the New York City Tuberculosis Registry, interviewing for tuberculosis history, skin testing, and chest radiography. Cases were classified as either previously or newly diagnosed. Among the 106 participants, 16 (15%) previously diagnosed tuberculosis cases were identified. Participants' tuberculosis histories were identified by the questionnaire (100%) or by registry match (69%). Eight participants (50%) were noncompliant with therapy. These findings prompted the establishment of a directly observed therapy program on site. PMID- 7485673 TI - Pet ownership among persons with AIDS in three Florida counties. AB - Interviews were conducted among 408 adults with acquired immunodeficiency syndrome at three local health departments to determine the proportion who owned pets, their perceived attachment to their pets, and the proportion who were informed about zoonoses. Nearly half (187, or 46%) were living with pets, most commonly dogs (64%), followed by cats (38%), fish (15%), birds (8%), reptiles (3%), and rodents (2%). Most pet owners (81%) reported an attachment to their pet. Only 10% were informed of zoonoses, albeit some incorrectly. Health care providers should recognize the high pet ownership rate among persons infected with human immunodeficiency virus and correctly inform their patients of strategies to sustain a low zoonotic disease incidence. PMID- 7485674 TI - AIDS phobia, public health warnings, and lawsuits: deterring harm or rewarding ignorance? AB - Courts in more than a dozen states have decided cases in which a person has claimed money damages for his or her fear of getting acquired immunodeficiency syndrome (AIDS). Although most courts have rejected such claims in the absence of actual exposure to the human immunodeficiency virus (HIV), their reasoning has varied slightly from state to state. This article argues that negligence law should not permit people who are HIV negative to recover damages for an unfounded fear of AIDS. Public health statements intended to educate the public about preventing HIV transmission may have encouraged some fear-of-AIDS lawsuits against health care practitioners. Although well intentioned, such statements have been used to justify inappropriate restrictions on medical practice and disclosure of a practitioner's HIV status. To avoid such misuse, such statements should be revised to make clear that the way in which procedures are performed, not who performs them, determines HIV transmission. PMID- 7485675 TI - The risk of domestic violence and women with HIV infection: implications for partner notification, public policy, and the law. AB - Partner notification has emerged as an important strategy in the fight against acquired immunodeficiency syndrome (AIDS), and states have now adopted a plethora of laws that encourage or mandate notification, often without the patient's consent. As human immunodeficiency virus (HIV) infection continues to spread among women, the future development of AIDS control strategies and public health laws must be shaped by concern for the safety and autonomy of patients who face a risk of domestic violence. Three distinct recommendations flow from this premise. First, all HIV-infected women should be assessed for the risk of domestic violence and offered appropriate interventions. Second, where a risk of abuse is indicated, partners should never be notified without the patient's consent. State laws that presently permit involuntary notification should be repealed or amended. Third, laws that punish a patient's refusal to notify partners should also be modified or repealed. PMID- 7485676 TI - Regulating controversial programs for unpopular people: methadone maintenance and syringe exchange programs. AB - One third of all cases of the acquired immunodeficiency syndrome (AIDS) in the United States are associated with the injection of illicit drugs. There is mounting evidence for the effectiveness of syringe exchange programs in reducing human immunodeficiency virus (HIV) risk behavior and HIV transmission among injection drug users. Expansion of syringe exchange would require increased public funding and undoubtedly would include government regulation of syringe exchanges. An analogy is drawn with the present system of regulation of methadone maintenance treatment programs and possible regulation of syringe exchange programs. Specific recommendations are offered to reduce the likelihood of repeating the regulatory problems of methadone maintenance treatment in future regulation of syringe exchange programs. PMID- 7485679 TI - Reevaluating the evidence on pesticide safety. PMID- 7485678 TI - The availability of disposable syringes in Russia. PMID- 7485677 TI - HIV risk factors and behaviors among men in a Milwaukee homeless shelter. PMID- 7485680 TI - Folic acid fortification for the prevention of neural tube defects: consensus needed on potential adverse effects. PMID- 7485681 TI - Insecticide-impregnated bed nets for malaria control: varying experiences from Ecuador, Colombia, and Peru concerning acceptability and effectiveness. AB - Between 1991 and 1994, an intervention program with permethrin- and lambdacyhalothrin-impregnated bed nets was carried out over a period of nine months in each of five endemic, malarious areas of Ecuador, Peru, and Colombia. This program was evaluated through household surveys, blood sampling, in-depth longitudinal studies, and entomologic analysis. Eighty-four communities (including approximately 35,000 individuals) were paired according to malaria incidence, size, and coverage with bed nets and then randomly allocated to intervention and control groups. The results showed that peoples' acceptance of the measure was related to their perception of an immediate protective effect against insects. The effectiveness of the bed nets, measured as a reduction of malaria incidence in intervention communities as against control communities, showed large variations between and within the study areas. The protective efficacy varied between 0% and 70% when looking only at the postintervention differences between intervention and control groups. The average protection was 40.8% when considering a four-month incidence of clinical malaria attacks and 28.3% when considering a two-week malaria incidence. Important factors for the success of the bed net program were insect susceptibility to pyrethroids, high coverage with impregnated bed nets, high malaria incidence, good community participation, high mosquito densities when people go to bed, and a high proportion of Plasmodium falciparum. In one area, where DDT spraying in the control communities was executed, the effectiveness of bed net impregnation was slightly better than that of spraying. PMID- 7485682 TI - Aedes aegypti control in the Lao People's Democratic Republic, with reference to copepods. AB - An evaluation of the Lao Aedes aegypti control program and of the predatory abilities of copepods from Vientiane, Lao People's Democratic Republic was undertaken before a field release of copepods in Thongkankam village, Vientiane. Copepods were transported to Australia for evaluation of predatory abilities and their survival under various nutrient and pH conditions. Mesocyclops guangxiensis was chosen for release over M. aspericornis due to its higher reproduction rate and its ability to survive in lower nutrient environments. Mesocyclops guangxiensis was released into 142 containers and 20 wells in a village in Vientiane. Copepods were present in 7% of the containers after one month and were absent six months postinoculation. In comparison, 100% of wells were still positive after six months, with average numbers of Ae. aegypti in the wells decreasing from 59.5 +/- 18.5 (+/- SEM) to 0 after six months. Numbers of Culex quinquefasciatus and Anopheles maculatus also decreased to 0 after six months. This study indicates that predacious copepods will be accepted by the community and could be integrated as a low-cost, persistent control agent into new strategies for improving surveillance and control of dengue vectors. PMID- 7485683 TI - Rift Valley fever virus in the cardia of Culex pipiens: an immunocytochemical and ultrastructural study. AB - In this paper, we consider the movement of Rift Valley fever (RVF) virus from infected mosquito midgut epithelial cells into the hemocoel as an important factor in the ultimate ability of the insect to transmit the virus. Our results are therefore significant in the context of vector competence. The mosquito Culex pipiens was identified as the primary vector of RVF in an epidemic that occurred in Egypt in the 1970s. On this basis, we have carried out several studies of RVF virus in this mosquito species. In the research reported here, we used immunocytochemical and transmission electron microscopic techniques to study the occurrence of RVF virus in the mosquito cardia and aspects of the histology and ultrastructure of this organ. The cardia is a complex organ consisting of both foregut and midgut tissue and is the location of the foregut-midgut junction. The cardia is of interest because it appears to provide routes of RVF virus egress from the midgut lumen and it is consistently infected in mosquitoes with disseminated infections, making it a potentially important site of viral amplification and an ideal site for studying RVF viral morphogenesis. In orally infected mosquitoes, large numbers of RVF virions were observed budding into the basal labyrinth associated with the outer cardial epithelial cells and into the noncellular matrix associated with the inner cardial epithelial cells and the cells of the intussuscepted foregut. In mosquitoes infected by injection of virus into the hemocoel and then held for different incubation periods, viral antigen was first detected in the cells of the intussuscepted foregut in the cardia and later in the cardial epithelial cells.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7485684 TI - Hepatitis B, C, D, and E markers in rural equatorial African villages (Gabon). AB - Serologic markers of hepatitis B virus (HBV), including HBV DNA, and of hepatitis delta virus were measured in three villages in Gabon. Of 303 subjects studied, 19% were carriers of hepatitis B surface antigen (HBsAg); 8.5% of these had anti delta antibodies. No difference among the three villages was observed. All HBV DNA carriers were children less than 11 years of age. In the 2-9-year-old group, 71% of the HBsAg-positive children tested were HBV DNA carriers. These results indicate that HBV prevalence is high in Gabon and emphasize the importance of horizontal transmission of HBV in Africa. Antibodies to hepatitis C virus, assessed in one of the three villages, were found with a prevalence of 35% with a second generation enzyme-linked immunosorbent assay (ELISA) and 24% with a third generation ELISA. None of 35 subjects tested for antibodies to hepatitis E virus was positive. PMID- 7485685 TI - Cow, farm, and ecologic risk factors of clinical vesicular stomatitis on Costa Rican dairy farms. AB - Twenty-two Costa Rican dairy herds were actively monitored for vesicular diseases between November 1991 and March 1992, in anticipation of the annual dry season occurrence of vesicular stomatitis (VS). Thirty-nine confirmed cases of VS were concentrated on nine farms, resulting in an average herd incidence rate of 11.1% of mature cattle for the affected farms, or 2.6% for all farms. Affected cattle were generally older (average age = 5.4 years), with seven-year-old cows having the highest age-specific incidence rate (6.5%). The New Jersey serotype of VS was diagnosed for all but four cases, primarily manifesting as teat vesicles. Cases had high acute and convalescent antibody titers for both major VS serotypes, New Jersey and Indiana, with significant titer increases after clinical disease. Screening and multiple variable regression analyses of 140 cow, farm, and ecologic variables revealed that clinical disease was significantly associated with cows being in lactation, and with higher acute antibody titers to serotype Indiana, but not with any predisposing diseases. Significant farm factors were the presence of poultry and a longer calving interval on the farm, but not higher rates of other diseases. According to a prior epidemiologic hypothesis, two ecologic factors were forced to be included into the farm model: the reported presence of sand flies, and a higher proportion of the farm being in forest land, with subsequent stepwise regression. The resulting model containing only the two forced variables was significant (P < 0.003) and correctly classified 78% of farms into the correct group, as compared with 79% for the final stepwise model.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7485686 TI - Vibrio cholerae in the horn of Africa: epidemiology, plasmids, tetracycline resistance gene amplification, and comparison between O1 and non-O1 strains. AB - The prevalence of Vibrio cholerae O1 and non-O1 has been investigated in numerous Somali regions of the Horn of Africa from 1983 to 1990. From January 1983 to January 1985 and between December 1986 and December 1990, no strains of V. cholerae O1 and 226 strains (5.3%) of V. cholerae non-O1 were isolated from 4,295 diarrhea cases. During a cholera epidemic in 1985 and 1986, the overall case fatality rate was 13% and the attack rate was 3-3.5 per 1,000 population. Matched case-control studies identified a waterborne route of transmission. A drug susceptible Ogawa strain from Ethiopia caused the introduction of the disease into northern Somalia. There were two major resistant derivatives of the original strain, and the one resistant to ampicillin, kanamycin, streptomycin, sulfonamide, and tetracycline (TC) predominated in the spreading disease. In 1986, susceptible Ogawa strains quickly displaced this resistant strain. The two incompatibility group C plasmids responsible for the resistance patterns had complex and scattered differences in their structures. Physical analysis of the plasmid DNA region coding for TC resistance demonstrated its genetic amplification in highly resistant variants of Ogawa strains. PMID- 7485687 TI - Short report: extended follow-up of the natural history of persons infected with Leishmania chagasi. AB - A five-year follow-up of the natural history of 924 children exposed to or infected with Leishmania chagasi was conducted in a rural area of northeast Brazil. Seventy-eight percent of the children sought were located. There was no evidence of smoldering disease or long subclinical latency in this population. The overall prevalence of clinical visceral leishmaniasis in this population was 6.1%, with a mortality rate of 10%. PMID- 7485688 TI - Biochemical systematics and population genetic structure of Anopheles pseudopunctipennis, vector of malaria in Central and South America. AB - An electrophoretic survey of 42 populations of Anopheles pseudopunctipennis collected throughout its known geographic distribution was performed to clarify the taxonomic status of this important malaria vector species. The results indicated strong differences in the allele frequencies of three enzyme loci (glycerol dehydrogenase, 6-phosphogluconate dehydrogenase, and phosphoglucomutase) of the 33 loci analyzed. No fixed electromorphic differences separate the populations of An. pseudopunctipennis. The populations of An. pseudopunctipennis showed little genetic divergence, with Nei distances ranging from 0 to 0.079. A comparison of An. pseudopunctipennis data with either one of three other Anopheles species showed a high genetic distance of 0.335 with a closely related species, An. franciscanus; 0.997 with An. crucians, and 2.355 with An. (Nyssorhynchus) albimanus. Geographic populations of An. pseudopunctipennis were classified into three clusters; one cluster included populations collected in North America (United States and Mexico) and Guatemala, one cluster included populations from Belize and South America (Colombia, Ecuador, Peru, Chile, and Argentina); and one cluster was represented by populations from the Island of Grenada (type-locality of An. pseudopunctipennis). Based on our isozyme analyses, we defined these clusters as three geographic populations of An. pseudopunctipennis. Of the two mainland populations, one extends from the southern United States south through Mexico and Guatemala, and the other extends north from southern South America through Central America to Belize. These two geographic populations converge in southern Mexico and northern Central America. One part of the convergence zone was identified in the area of eastern Guatemala and southern Belize. PMID- 7485689 TI - Short report: peak period of filarial transmission. AB - This study was designed to determine the peak period of infective mosquito biting and thus the greatest potential for filarial transmission to occur. It was found that biting density, natural infection, and infectivity rates of Culex quinquefasciatus were significantly higher in the third quadrant of the night (from midnight to 3:00 AM) than at other times. This was true in both an urban and a rural environment. Avoidance of mosquito bite by any means during this time period might reduce and limit filarial transmission to a great extent. PMID- 7485690 TI - Bubonic leishmaniasis: a common manifestation of Leishmania (Viannia) braziliensis infection in Ceara, Brazil. AB - Enlarged regional lymph nodes have been reported to accompany the cutaneous lesions of Leishmania (Viannia) braziliensis (= L. braziliensis). A survey in Ceara State, Brazil indicated that 77% of persons (456 of 595) with parasitologically confirmed cutaneous leishmaniasis reported lymphadenopathy in addition to skin lesions. A group of 169 persons with recently diagnosed leishmaniasis and lymph nodes measuring > or = 2 cm in diameter (mean = 3.6 cm, maximum = 10.5 cm) underwent detailed clinical examination. Lymphadenopathy preceded the skin lesions in more than two-thirds of these, on the average by two weeks. Cultures of lymph node aspirates yielded Leishmania more frequently (86%) than cultures of aspirates of skin (53%) or biopsies of skin (74%). Parasites were isolated from the peripheral blood of one patient. Persons with lymphadenopathy gave a history of fever and had enlarged livers or spleens more often than a comparison group of 50 persons with cutaneous lesions but no lymphadenopathy. Persons with lymphadenopathy had more intense leishmanin skin reactions and lymphocyte proliferation following stimulation with specific antigens, whereas persons without lymphadenopathy had a higher frequency of previous infection. Isolates of parasites from both groups were identified as L. braziliensis. These data demonstrate the early spread of L. braziliensis beyond the skin and suggest differences in host immunity between persons with and without lymphadenopathy. Leishmaniasis braziliensis should be considered in cases of unexplained lymphadenopathy in endemic areas. PMID- 7485691 TI - Short report: the spatula test: a simple bedside test to diagnose tetanus. AB - Four hundred patients with suspected tetanus were studied to determine the value of the spatula test to diagnose tetanus. A positive test result (reflex spasm of the masseters on touching the posterior pharyngeal wall) was seen in 359 (94%) of 380 patients with tetanus and in no patient without tetanus. Thirty-three of 400 patients (13 with tetanus and 20 with other diagnoses) had a negative test result (a gag reflex with attempted expulsion of the spatula). Thus, the test performed on presentation had a high specificity (100%) and sensitivity (94%) for diagnosing tetanus. PMID- 7485692 TI - In vitro activity of atovaquone against the African isolates and clones of Plasmodium falciparum. AB - The in vitro activity of atovaquone (566C80) was evaluated and compared with that of chloroquine, quinine, mefloquine, halofantrine, artemether, pyrimethamine, and cycloguanil against African isolates and clones of Plasmodium falciparum using an isotopic, semimicro, drug susceptibility test. Atovaquone was highly active against the chloroquine-susceptible L-3 (geometric mean 50% inhibitory concentration [IC50] = 0.978 nM) and L-16 clones (mean IC50 = 0.680 nM) and against the multidrug-resistant FCM 29 clone (mean IC50 = 1.76 nM). Similar low IC50 values for atovaquone were observed against the chloroquine-susceptible isolates (n = 35; geometric mean IC50 = 0.889 nM) and the chloroquine-resistant parasites (n = 26; geometric mean IC50 = 0.906 nM). The in vitro responses between atovaquone and the other antimalarial drugs were not correlated, indicating the absence of in vitro cross-resistance. The high in vitro activity of atovaquone without any in vitro evidence for cross-resistance with other antimalarial drugs against the naturally occurring malaria parasites is a factor that favors further development of the drug for clinical use. PMID- 7485693 TI - Studies on cryopreservation of Toxoplasma gondii and its antigenicity in mice. AB - Two strains (RH and GC, the latter of which is a Taiwan isolate of porcine origin) of Toxoplasma gondii were kept at -20 degrees C, -60 degrees C, and in liquid nitrogen (-196 degrees C) to follow the time course change in viability and virulence of the parasites by direct count and animal inoculation methods. Changes in antibody titers in some of the mice inoculated with the thawed organisms were assayed by the indirect immunofluorescent antibody test. Viability and virulence of T. gondii were best preserved by storage in liquid nitrogen. Tachyzoites kept in liquid nitrogen for eight years still can lead to the death of the injected mice in 2-3 weeks. Virulence of the tachyzoites could be maintained for eight weeks at most at -20 degrees C and -60 degrees C. Dimethylsulfoxide (DMSO) seemed to be a better cryoprotectant for T. gondii than glycerol, but the DMSO-preserved organisms resulted in fewer tachyzoite containing peritoneal exudates in inoculated mice than the glycerol-preserved organisms. The local isolate (GC strain) tachyzoites tolerated cryopreservation less well than the RH strain parasites. Low antibody titers (at most 1:64) were produced in mice that survived more than 16 days after inoculation with thawed tachyzoites. PMID- 7485694 TI - Growth and migration of Borrelia burgdorferi in Ixodes ticks during blood feeding. AB - We have studied the growth of Borrelia burgdorferi in nymphal ticks (Ixodes scapularis) feeding on mice using confocal fluorescence microscopy to follow the distribution of spirochetes. In starved nymphs, the bacteria were only detected in the midgut and each nymph had a mean of 496 spirochetes. Upon attachment of nymphs to the host, the bacteria grew with a doubling time close to 4 hr and reached a mean of 7,848 spirochetes per nymph 15 hr after attachment. During this initial period (36 hr) of rapid growth, the bacteria appeared to be restricted to the gut, but after 48 hr, the spirochetes had disseminated to the salivary glands in the majority of nymphs examined. Thus, a critical event that allows the spirochetes to disseminate and infect the salivary glands takes place 36-48 hr after attachment. A maximum number of 166,575 spirochetes per nymph was noted 72 hr after attachment. Soon after completion of feeding and detachment from the host (96 hr), the mean number of spirochetes decreased to 95,410 per nymph and the spirochetes appeared to be cleared from organs other than the midgut. Thus, dissemination of spirochetes within the vector appears to be a transient phenomenon. These results provide strong evidence in favor of a salivary route of disease transmission while also demonstrating the utility of confocal microscopy to study vector-pathogen interactions in general. PMID- 7485695 TI - Characterization of clone 13, a naturally attenuated avirulent isolate of Rift Valley fever virus, which is altered in the small segment. AB - The 74HB59 strain of Rift Valley fever (RVF) virus, isolated from a human case in the Central African Republic, was shown to be composed of a heterogeneous population of viruses when plaque-purified clones were analyzed for their reactivity with monoclonal antibodies (MAbs) directed against the nucleocapsid (N) protein or the nonstructural (NSs) protein. One of these clones, C13, was of particular interest in that it proved to be avirulent in mice and hamsters, and highly immunogenic. Although C13 showed normal reactivity with a large panel of MAbs directed at the glycoproteins, it failed to react with specific MAbs or polyclonal antibodies directed at the NSs protein and with a specific MAb recognizing the N protein of the Egyptian strains. Consequently, the small RNA segment, which encodes the N and NSs proteins in an ambisense strategy, was sequenced and compared with the existing sequence of the attenuated MP-12 RVF virus strain. We found that the NSs gene contained, in addition to two conservative coding changes, a large internal deletion of 549 nucleotides that removes 69% of the open reading frame but conserves in-frame the N and C termini of the predicted translation product. In addition, the sequence revealed that the N protein of C13 contained a single amino acid change. Clone C13 replicated normally in certain cell types in vitro and in Culex pipiens mosquitoes after intrathoracic inoculation, but established abortive infections in MRC-5 human fibroblasts. PMID- 7485696 TI - In vivo proviral burden and viral RNA expression in T cell subsets of patients with human T lymphotropic virus type-1-associated myelopathy/tropical spastic paraparesis. AB - We used in situ hybridization combined with immunocytochemistry, cell sorting, and the polymerase chain reaction (PCR) to investigate clinical events in three asymptomatic carriers of human T lymphotrophic virus type-1 (HTLV-1) and ten patients with HTLV-1-associated myelopathy/tropical spastic paraparesis (HAM/TSP). The objective was to determine which T cell subset of peripheral blood mononuclear cells (PBMC), CD4 or CD8, were infected by HTLV-1 and the manner in which HTLV-1 proviral DNA was expressed at the level of the single cell. Both CD4 positive and CD8-positive cells of the PBMC from five patients with HAM/TSP were infected with HTLV-1. The proportion of HTLV-1-infected cells was 2.5-40% in the CD4-positive subset and 1.0-65% in the CD8- positive subset, when quantified by PCR using HTLV-1-infected MT2 cells as a positive standard. Proviral DNA of HTLV 1 was expressed in both CD4-positive cells and CD8-positive cells of the PBMC from six patients with HAM/TSP and three asymptomatic HTLV-1 carriers. In patients with HAM/TSP, the proportion of the cells expressing HTLV-1 proviral DNA was 0.02-0.1% in both subsets. In asymptomatic carriers, the expression of HTLV-1 proviral DNA was 0.01-0.02% in the CD4-positive subset and 0.01% in the CD8 positive subset. Therefore, HTLV-1 possessed similar in vivo cellular tropism for both CD4-positive cells and CD8-positive cells and HTLV-1 proviral DNA was expressed in vivo in both circulating T cell subsets.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7485697 TI - Comparative study of the immunogenicity and safety of two dosing schedules of hepatitis B vaccine in neonates. AB - Healthy Egyptian neonates born to hepatitis B surface antigen (HBsAg) seronegative mothers were randomly enrolled in one of three vaccination schedules. A dose of 2.5 micrograms of recombinant HB vaccine was given at birth, two, and six months of age (group A) or two, four, and nine months of age (group B). These two groups and a third control group (group C) also were given the other routine childhood vaccines (BCG, DPT, polio, and measles). Blood samples were taken one month after the third vaccine dose in groups A (seven months of age) and B (10 months of age), and a second follow-up blood sample was taken at the age of 18 months for all three groups. Sera were tested for HBsAg and antibody to hepatitis B core antigen, and quantitatively for antibody to hepatitis B surface antigen (anti-HBs) using commercial enzyme immunoassay kits. The vaccine was well tolerated and side effects were limited to local soreness, redness, or temporary swelling. Among 590 infants who were followed-up, good (51 300 mIU anti-HBs/ml) or excellent (> 300 mIU/ml) immune responses occurred in 85% of the infants in group A and in 96% in group B. Geometric mean titers of anti HBs at the first and second follow-up were 306 and 55 mIU/ml in group A, and 1,492 and 147 mIU/ml in group B. The recombinant HB vaccine is safe and immunogenic when given in three doses of 2.5 micrograms in either regimen, but delay of the booster dose of the vaccine until nine months after birth produced a higher immune response. PMID- 7485698 TI - Safety, immunogenicity, and pilot efficacy of Plasmodium falciparum sporozoite and asexual blood-stage combination vaccine in Swiss adults. AB - This study was part of a larger program to develop a vaccine effective against Plasmodium falciparum infection caused by sporozoites and clinical malaria caused by asexual blood stages. In a phase 1 study of safety and immunogenicity, two recombinant proteins (Ro 46-2717, a circumsporozoite [CS] protein) construct with a molecular mass of 35 kD, and Ro 46-2924, a merozoite surface antigen [MSA-2] construct with a molecular mass of 25 kD) adsorbed onto alum were injected in two low (20 micrograms) or two high (100 micrograms) doses in the right and left deltoid muscles of 33 healthy Swiss volunteers; six other volunteers received a placebo (alum alone). Twenty-six participants reported 51 immunization-related adverse events, mainly pain at the injection site. Mean antibody titers to CS protein and MSA-2 in an indirect immunofluorescence assay peaked four weeks after the second immunization without evidence of boosting (i.e., sharp increase in titer). By that time, 56% and 31% of the vaccinees seroconverted to CS protein and MSA-2, respectively, with the increase in MSA-2 titer being weaker than that for the CS protein. After a third immunization, five vaccinees volunteered to be challenged by three or four infective bites of Anopheles stephensi. Prepatent and incubation periods in all five were comparable with unvaccinated historic controls challenged under similar conditions, and all had symptoms of clinical falciparum malaria. We conclude that the vaccine components were safe and immunogenic but there was no evidence that this immunization regimen with the CS protein plus MSA-2 component was able to prevent infection.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7485699 TI - Murine immunoglobulin G subclass responses following immunization with live dengue virus or a recombinant dengue envelope protein. AB - Murine immunoglobulin G (IgG) subclass responses to immunization are restricted to certain subclasses depending on the nature of the immunogen. Immunization with live viruses generally leads to a predominant IgG2a response, which may be the most effective at resisting future challenge due to the unique effector functions of IgG2a. Knowledge of subclass responses following immunization with dengue vaccine candidates may be helpful in determining which candidates are most efficacious. We measured the dengue-specific IgG subclass responses of BALB/c mice following immunization with live dengue-2 virus or with a partially purified recombinant dengue-2 envelope (E) protein. Subclass responses following immunization with live virus were IgG2a > IgG1 > IgG2b > IgG3, as opposed to IgG1 > IgG2a > IgG2b > IgG3 after immunization with recombinant protein. Responses of all subclasses except IgG1 were greater following immunization with live dengue than with the recombinant E protein. Neutralizing antibody titers were also higher after immunization with live virus than with E protein and were positively correlated with dengue-specific IgG2a responses in mice immunized with recombinant E protein. Following separation of the four IgG subclasses by chromatography, the IgG2a fraction exhibited the greatest neutralizing activity. The results seen after immunization with live dengue virus or recombinant E protein in this study are in concordance with studies involving other viruses and viral proteins and may have implications for the development of an effective vaccine for dengue. PMID- 7485700 TI - Seroprevalence of cysticercosis in an Orthodox Jewish community. AB - Neurocysticercosis cases were identified in 1991 in an Orthodox Jewish community. Transmission was linked to tapeworm-infected immigrant housekeepers from countries where Taenia solium is endemic. To evaluate the extent of and risks for locally acquired cysticercosis, a seroprevalence survey was conducted in 9% of the households in this community. Cysticercosis antibodies were detected in 23 (1.3%) of 1,789 persons from 612 families. All 23 seropositive persons were asymptomatic, and no intracerebral lesions were found for the 21 seropositive persons who underwent brain imaging. Seropositivity was associated with female sex (relative risk [RR] = 2.45, P = 0.049), hiring a domestic worker for child care duties (RR = 3.79, P = 0.05), and with employees from Central America (RR = 2.70, P = 0.0001). Exposure to T. solium in this community is unexpectedly high. Widespread employment of domestic workers from endemic regions and high employee turnover contributes to exposure risk. PMID- 7485701 TI - Evaluation of risk factors for house infestation by Triatoma infestans in Brazil. AB - An active entomologic survey was conducted by a team of trained health workers in a rural area endemic for Chagas' disease in central Brazil. They used pyrethrum as a flushing agent and 4,232 houses were inspected for triatomine bugs both inside and in the immediate environs. Houses with Triatoma infestans or evidence of an established colony were identified and defined as infested houses (cases). The building and environmental characteristics of 161 randomly selected infested houses were compared with 161 matched, noninfested houses (controls) that were the shortest distance from the infested house. Domestic and peridomestic potential risk factors associated with house infestation by Triatoma infestans were assessed by logistic regression analysis. Incomplete house construction (odds ratio [OR] = 2.5, 95% confidence interval [CI] = 1.5-4.1) was confirmed as a risk factor related to the presence or evidence of Triatoma infestans in the dwellings. The study also disclosed a statistically significant association between the presence of rats (OR = 1.6, 95% CI = 1.1-2.6) and indoor crop storage (OR = 2.3, 95% CI = 1.1-5.2) and house infestation. Further experimental field studies using tagged rodents should be conducted to assess their epidemiologic role in the domestic chain of Trypanosoma cruzi transmission. PMID- 7485702 TI - Human Toxoplasma infection in Kuna and Embera children in the Bayano and San Blas, eastern Panama. AB - We conducted a survey of 760 Amerindian children 2-12 years of age in the Bayano and San Blas areas of Panama in 1991 to determine the prevalence of serum antibodies to Toxoplasma gondii and the importance of hypothesized risk factors in human-induced native and sylvatic conditions, which have had few environmental changes, as opposed to rural and urban areas in Panama previously studied. The overall prevalence of infection ranged between 0% and 42.5%. No age curve was detected, indicative of nonconstant transmission. Only two hypothesized risk factors, floor type and having cats inside the house, were significantly associated with the presence of antibodies in some of the communities. Antibody prevalence appeared to be associated more with the community of residence than with any specific behavior. The risk factor of importance may be the level of oocyst contamination, since infection by tissue cysts in meat was excluded. On three of the nine islands studied, no antibody was detected in the children or the cats. It would appear that T. gondii is not present on these islands. Although the data did not support the importance of many of the hypothesized risk factors, the study is consistent with the theory of transmission by oocysts and the importance of cats in transmission. PMID- 7485703 TI - Transmission of Toxoplasma gondii in Panama City, Panama: a five-year prospective cohort study of children, cats, rodents, birds, and soil. AB - A cohort of more than 500 children from Panama City, Panama was studied prospectively over five years for acquisition of antibody to Toxoplasma gondii. The direct agglutination test showed that 72 of 571 children seroconverted between one and six years of age, for a cumulative incidence of 12.6%. Children were examined by pediatricians quarterly, and illnesses that had occurred in the interval and their activities were noted on questionnaires. Thirty-eight variables were examined for their role as risk factors for seroconversion. There was a higher correlation between children's seroconversion and contact with dogs than with cats. Combinations of significant predictors without dogs explained only 67% of the seroconversions, but the same factors with dogs explained 90%. On the other hand, ingestion of raw or rare meat or eggs appeared to play no role in transmission. Cats were examined and 110 (45.6%) of 241 had Toxoplasma antibody on the first bleeding. Only two (0.5%) of 383 cat fecal specimens, when tested in mice, resulted in seroconversion. Ten (1.1%) of 924 soil samples resulted in seroconversion in mice that had been injected. Antibody to Toxoplasma was found in 52 (23.3%) of 226 rats (Rattus norvegicus) and two (0.035%) of 571 mice (Mus musculus). Two hundred sixteen birds of 16 different species were bled. Antibody to Toxoplasma was found in 13.4% of these birds, mostly in grackles, blue-gray tanagers, and doves. The rate of isolation of Toxoplasma was low: one of 23 in rats and three of 201 in birds. High relative risks (RRs) of transmission to children were predicted by contact histories with nursing dogs (RR = 5.8), weaned dogs (RR = 4.7), many flies (RR = 3.6), 6-12-month-old dogs (RR = 3.4), weaned cats (RR = 3.0), 6-12-month-old cats (RR = 2.7), nursing cats (RR = 2.5), much garbage (RR = 2.4), and many roaches (RR = 2.2). The high statistical correlation of dog contact with seroconversion in children suggests the possibility that dogs, by eating and rolling in cat feces, are instrumental in mechanically transmitting Toxoplasma infection. In addition, flies, and to a lesser extent, cockroaches, may have practically important roles in transmission. PMID- 7485704 TI - Variation in ribosomal DNA intergenic spacers among populations of Anopheles albimanus in South and Central America. AB - Variation in the length and copy number of intergenic spacers (IGS) of nuclear ribosomal DNA were examined to test for genetic differentiation among Anopheles albimanus populations. Extensive collections were made in Guatemala but populations were also sampled over a large range of its distribution in Central and South America. Discriminant analysis of IGS patterns in individual mosquitoes indicated that populations generally had unique sets of IGS length variants. The IGS patterns from populations on the Pacific side of Central America were distinct from those on the Atlantic side or from South America. Cluster analysis indicated a similar trend. The IGS diversity in Central America was 50% greater than in South America. These results suggest that barriers to gene flow exist among Atlantic and Pacific coast populations of An. albimanus. No gene flow barriers were detected among populations from Colombia, Ecuador, Peru, and Venezuela. PMID- 7485706 TI - Predictions of adult Anopheles albimanus densities in villages based on distances to remotely sensed larval habitats. AB - Remote sensing is particularly helpful for assessing the location and extent of vegetation formations, such as herbaceous wetlands, that are difficult to examine on the ground. Marshes that are sparsely populated with emergent macrophytes and dense cyanobacterial mats have previously been identified as very productive Anopheles albimanus larval habitats. This type of habitat was detectable on a classified multispectral System Probatoire d'Observation de la Terre image of northern Belize as a mixture of two isoclasses. A similar spectral signature is characteristic for vegetation of river margins consisting of aquatic grasses and water hyacinth, which constitutes another productive larval habitat. Based on the distance between human settlements (sites) of various sizes and the nearest marsh/river exhibiting this particular class combination, we selected two groups of sites: those located closer than 500 m and those located more than 1,500 m from such habitats. Based on previous adult collections near larval habitats, we defined a landing rate of 0.5 mosquitoes/human/min from 6:30 PM to 8:00 PM as the threshold for high (> or = 0.5 mosquitoes/human/min) versus low (< 0.5 mosquitoes/human/min) densities of An. albimanus. Sites located less than 500 m from the habitat were predicted as having values higher than this threshold, while lower values were predicted for sites located greater than 1,500 m from the habitat. Predictions were verified by collections of mosquitoes landing on humans. The predictions were 100% accurate for sites in the > 1,500-m category and 89% accurate for sites in the < 500-m category. PMID- 7485705 TI - Discrimination of all members of the Anopheles punctulatus complex by polymerase chain reaction--restriction fragment length polymorphism analysis. AB - A method has been developed to identify the members of the Anopheles punctulatus complex using polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP). Members of the An. punctulatus complex are the most important vectors of malaria in the southwest Pacific and consist of 10 cryptic species, An. farauti no. 1-7, An. punctulatus, An. sp. near punctulatus, and An. koliensis. For each species, PCR amplification of the ribosomal DNA internal transcribed spacer produced a 750-basepair product. Digestion with Msp I and electrophoresis on a 3.0% agarose gel results in banding patterns unique to each species. Isolates of the same species from different locations gave an identical pattern. The technique is sensitive enough so that a PCR-RFLP can be generated from as little as a single mosquito leg, allowing the rest of the mosquito to be used for other important epidemiologic analyses such as determining host feeding source, and for parasite detection. PMID- 7485707 TI - A simulation model of the epidemiology of urban dengue fever: literature analysis, model development, preliminary validation, and samples of simulation results. AB - We have developed a pair of stochastic simulation models that describe the daily dynamics of dengue virus transmission in the urban environment. Our goal has been to construct comprehensive models that take into account the majority of factors known to influence dengue epidemiology. The models have an orientation toward site-specific data and are designed to be used by operational programs as well as researchers. The first model, the container-inhabiting mosquito simulation model (CIMSiM), a weather-driven dynamic life-table model of container-inhabiting mosquitoes such as Aedes aegypti, provides inputs to the tranmission model, the dengue simulation model (DENSiM); a description and validation of the entomology model was published previously. The basis of the transmission model is the simulation of a human population growing in response to country- and age-specific birth and death rates. An accounting of individual serologies is maintained by type of dengue virus, reflecting infection and birth to seropositive mothers. Daily estimates of adult mosquito survival, gonotrophic development, and the weight and number of emerging females from the CIMSiM are used to create the biting mosquito population in the DENSiM. The survival and emergence values determine the size of the population while the rate of gonotrophic development and female weight estimates influence biting frequency. Temperature and titer of virus in the human influences the extrinsic incubation period; titer may also influence the probability of transfer of virus from human to mosquito. The infection model within the DENSiM accounts for the development of virus within individuals and its passage between both populations. As in the case of the CIMSiM, the specific values used for any particular phenomenon are on menus where they can be readily changed. It is possible to simulate concurrent epidemics involving different serotypes. To provide a modicum of validation and to demonstrate the parameterization process for a specific location, we compare simulation results with reports on the nature of epidemics and seroprevalence of antibody in Honduras in low-lying coastal urbanizations and Tegucigalpa following the initial introduction of dengue-1 in 1978 into Central America. We conclude with some additional examples of simulation results to give an indication of the types of questions that can be investigated with the models. PMID- 7485708 TI - Comparison of a new ovine antigen binding fragment (Fab) antivenin for United States Crotalidae with the commercial antivenin for protection against venom induced lethality in mice. AB - Snake venom poisoning is a medical emergency requiring immediate attention and the exercise of considerable judgment. Of the estimated 8,000 bites inflicted by venomous snakes in the United States each year, approximately 6,000 are treated with commercial antivenin. The only commercially available antivenin for North American Crotalidae envenomation is Antivenin (Crotalidae) Polyvalent (equine origin) (ACP; Wyeth Laboratories, Philadelphia, PA). A common complication is the high incidence of hypersensitivity reactions, occurring in more than 75% of patients treated with ACP. To minimize these side effects, a novel, affinity purified, antigen binding fragment (Fab) antivenom (FabAV) for Crotalidae venom poisoning has been produced from the sera of sheep. The new product is Antivenin Polyvalent Crotalid (Ovine) Fab (Crotab; Therapeutic Antibodies, Inc., Nashville, TN). The current report compares the potencies in mice of FabAV and ACP against venom-induced lethality. The results indicate that FabAV is 3.1-9.6 times more potent than ACP for the prevention of lethality of the nine United States venoms tested. For one of the venoms, Crotalus viridis helleri, FabAV was efficacious while ACP was not. PMID- 7485709 TI - Compliance with antimalarial chemoprophylaxis and the subsequent development of malaria: a matched case-control study. AB - To determine if there is a difference in compliance with antimalarial chemoprophylaxis between febrile travelers with and without malaria, 157 patients with malaria, a history of fever, and recent travel to a malaria-endemic area were compared with 157 matched controls. Antimalarial prophylaxis had been taken by 48% of all patients. Chemoprophylaxis use was correlated with region and purpose of travel. Cases were less likely to have taken prophylaxis (53%) than controls (76%) (odds ratio = 0.35, confidence interval = 0.27, 0.73), even after controlling for region of travel, purpose of travel, and previous exposure to malaria. Chemoprophylaxis was effective in reducing malaria risk. Travel agents and health practitioners should provide all travelers to malaria-endemic areas with adequate information about chemoprophylaxis and its importance. PMID- 7485710 TI - Diagnosis of malaria in the febrile traveler. AB - To determine if clinical presentation can be used for predicting malaria infection in febrile patients with recent travel to a malaria-endemic area, 157 patients with malaria, a history of fever, and recent travel to a malaria-endemic area were compared with 157 matched controls in case-control study. Fever pattern, symptom duration, temperature, splenomegaly, and platelet count were correlated with malaria infection. These criteria, however, either singly or in combination, had low sensitivity for accurately identifying patients with malaria. Because no criterion could accurately predict the presence of malaria, it is concluded that microscopic examinations of blood for malaria parasites should be done in all symptomatic patients with a history of travel to a malaria endemic area. PMID- 7485711 TI - Oral artesunate in the treatment of uncomplicated hyperparasitemic falciparum malaria. AB - Patients with uncomplicated hyperparasitemic falciparum malaria are usually given parenteral antimalarial treatment to prevent a progression to vital organ dysfunction and death. Since the oral artemisinin derivatives are more rapidly effective than other antimalarial drugs, we compared oral artesunate (4 mg/kg/day for three days with mefloquine 25 mg/kg on the second day) with an intravenous quinine loading dose (20 mg of salt/kg initially then 10 mg/kg every 8 hr, followed by mefloquine 25 mg/kg) in an open paired randomized trial in 60 patients with acute falciparum malaria and greater than 4% parasitemia, but no evidence of vital organ dysfunction. There were no deaths and none of the patients progressed to develop severe malaria. Oral artesunate treatment resulted in shorter median [range] times to fever clearance (19 hr [4-45] versus 47 hr [4 107]) (P < 0.0001), parasite clearance (36 hr [18-61] versus 82 hr [36-104]) (P < 0.0001), and discharge from the hospital (25 hr [12-44] versus 58 hr [24-115]) (P < 0.0001). There was no toxicity attributable to artesunate. The cure rates by day 28 were 70% (19 of 27) and 39% (11 of 27) in the artesunate and quinine groups, respectively (relative risk = 1.7; 95% confidence interval = 1.0-3.0). Oral artesunate was simpler, cheaper, safer, and more effective than intravenous quinine for the treatment of uncomplicated hyperparasitemia. PMID- 7485712 TI - In vitro susceptibility of Plasmodium falciparum to chloroquine, amodiaquine, quinine, mefloquine, and sulfadoxine/pyrimethamine in Equatorial Guinea. AB - Between March 1990 and June 1992, a study was carried out in Equatorial Guinea on the in vitro response of Plasmodium falciparum to different antimalarial drugs. Field work for the study was conducted both in the country's island region as well as on the mainland, and resistant isolates were found to exhibit interregional differences. On the island of Bioko, 204 tests were performed with 16% (11 of 69) resistant to chloroquine, 9% (4 of 46) resistant to quinine, 14% (6 of 43) resistant to a combination of sulfadoxine/pyrimethamine, and 6.5% (3 of 46) resistant to amodiaquine. In the mainland area of Bata, the same antimalarial drugs and mefloquine were tested with the following results: 9% (5 of 58) resistant to chloroquine; 2% (1 of 58) resistant to amodiaquine, and 3% (2 of 58) resistant to a combination of sulfadoxine/pyrimethamine. No isolates resistant to quinine or mefloquine were found. Effective concentrations (EC50, EC90, and EC99) and regression lines (log dose/response) for each antimalarial drug were calculated to establish a surveillance system for antimalarial drug chemosensitivity in Equatorial Guinea. Finally, 12 isolates from 12 patients previously treated with chloroquine were studied to compare both tests (in vivo in vitro) and obtain a correlation between the RII and RIII types of in vivo and in vitro resistances. No correlation for the RI type was found between the two methods. PMID- 7485713 TI - Prediction of accelerated cure in Plasmodium falciparum malaria by the elevated capacity of tumor necrosis factor production. AB - Cytokine regulation was compared in three groups of Gabonese patients with Plasmodium falciparum malaria before and after therapy; adults with uncomplicated malaria, children with uncomplicated malaria, and children with severe malaria. Plasma levels of tumor necrosis factor (TNF), interleukin-6 (IL-6), IL-8, TNF receptors (TNF R), and the TNF/TNF R ratios were significantly higher in severe malaria compared with uncomplicated malaria. High plasma levels of all immunoregulatory molecules were associated with slow cure after therapy. In all patients, phytohemagglutinin-induced cytokine production was depressed on admission compared with convalescence. A significant difference was the higher TNF production capacity in patients with severe malaria on day 2 and day 5 compared with that in patients with uncomplicated malaria. In contrast to IL-6 and IL-8, a high TNF production capacity during the acute phase of malaria predicted a rapid clinical and parasitologic cure in the patients. These findings illustrate the dual role of TNF in the protection and pathology of malaria. PMID- 7485714 TI - Correlation of high serum levels of tumor necrosis factor-alpha with disease severity in human African trypanosomiasis. AB - The levels of tumor necrosis factor-alpha (TNF-alpha) in sera from Trypanosoma brucei gambiense-infected patients from the endemic region of Boko Songho (Bouenza focus in Congo) were measured. An increase was observed in sera from patients (geometric mean = 53.75 pg/ml, n = 69) compared with control subjects from the same endemic area (6.72 pg/ml, n = 31). The patients were classified as being in the early (blood lymphatic) stage and late (meningo-encephalitic) stage of disease according to the presence of parasites and cells in cerebrospinal fluid (CSF). An increase in TNF-alpha was noted in late stage patients (68.42 pg/ml, n = 28) compared with early stage patients (43.68 pg/ml, n = 41). Those patients with fever, asthenia, and edema and those with neurologic signs had higher levels of TNF-alpha (89.36 pg/ml, n = 26) than others (38.07 pg/ml, n = 43). No differences in TNF-alpha levels were seen when trypanosomes were detected in one location (blood, lymph nodes, or CSF) or two or three locations. These data show that the levels of TNF-alpha in serum of T. b. gambiense-infected patients were correlated with disease severity (presence of signs of inflammation or presence of major neurologic signs) and indicate that TNF-alpha could be involved in some aspects of human African trypanosomiasis physiopathology. PMID- 7485715 TI - A correlation between sequestered parasitized erythrocytes in subcutaneous tissue and cerebral malaria. AB - Sequestered parasitized erythrocytes were found in microvessels of subcutaneous tissues in a comatose patient with cerebral malaria even though the blood smears were negative after quinine treatment. This situation reflects the continued presence of sequestered parasites in the brain and suggests that negative parasitemia in peripheral blood does not necessarily mean the end point of malarial treatment has been reached. Our findings suggest that biopsy of subcutaneous tissue from severe malaria patients may be useful for determining the severity and prognosis of malaria patients. PMID- 7485717 TI - Neurocysticercosis and performance on neuropsychologic tests: a family study in Ecuador. AB - Neurocysticercosis (NCC) is a common neurologic disorder in developing countries, where it may pose a major public health challenge. Recently, the disorder has become more commonly diagnosed in developed countries as a result of the influx of migrants from countries where the disease is endemic. The clinical syndrome associated with NCC includes neurologic, physical, and functional problems. Since the locus of the infection is mainly the central nervous system, there is a risk of neuropsychologic dysfunction. This study was conducted in Ecuador in a group of 123 subjects (49 males and 74 females, 9-62 years of age) from a community sample that was part of a larger neuroepidemiologic inquiry. A discriminant function procedure was used to select the tests that would be most sensitive at distinguishing between affected and nonaffected individuals. The results suggest that behavioral functions that include aspects of inhibitory control, motor, and visual-motor output are impaired in adolescent and adult subjects with NCC. PMID- 7485716 TI - Increased erythropoietin production in children with severe malarial anemia. AB - Plasma immunoreactive erythropoietin concentrations were determined in 84 children with Plasmodium falciparum malaria in Gabon. There was an inverse log/linear relationship between hemoglobin or hematocrit and plasma erythropoietin, indicating that erythropoietin levels increased exponentially as circulating hemoglobin decreased. These result show that P. falciparum malaria does not lead to decreased erythropoietin production, and in turn reduced erythropoietin production does not contribute to the pathogenesis of malarial anemia. There is an adequate response of erythropoietin to anemia in children with P. falciparum malaria. PMID- 7485718 TI - Immunodiagnosis of polycystic hydatid disease/polycystic echinococcosis due to Echinococcus vogeli. AB - A crude antigenic metacestode extract from Echinococcus vogeli was assessed by enzyme-linked immunosorbent assay (ELISA) and showed strong binding activity with serum antibodies from patients with polycystic echinococcosis. Major cross reactions occurred with serum antibodies from patients with cystic and alveolar echinococcosis and from patients infected with other species of helminths. An E. vogeli antigen fraction, Ev2, was subsequently purified by immunosorption. The respective Ev2 ELISA demonstrated improved specificity, allowing discrimination of non-Echinococcus infections from polycystic echinococcosis. Based upon the calculation of a comparative (Ev-crude ELISA versus Ev2 ELISA) reactivity index, it became possible to discriminate all cystic echinococcosis cases, but only some alveolar echinococcosis cases, from polycystic echinococcosis. Immunoblot analyses revealed an antibody banding pattern highly conserved among polycystic, cystic, and alveolar echinococcosis. However, immunoblotting reliably distinguished between echinococcosis and all non-Echinococcus infections. PMID- 7485719 TI - Differential reactivity of immune sera from human vaccinees with field strains of eastern equine encephalitis virus. AB - Eastern equine encephalitis (EEE) virus is a mosquito-borne alphavirus that can produce a severe and often fatal acute encephalitis in humans, with significant neurologic sequelae in survivors. Due to the serious nature of the disease, an investigational inactivated EEE vaccine (PE-6) is available to individuals at risk for infection. Both serologic and recent molecular analyses of EEE viruses have demonstrated marked differences between the two antigenic varieties of EEE virus, designated North American (NA) and South American (SA). In view of these findings, we have examined the reactivity of sera from three individuals immunized with the EEE vaccine, derived from an NA isolate, with field strains of EEE virus. Anti-EEE serum antibodies from vaccinees reacted strongly in Western blot assays with both of the envelope (E1 and E2) glycoproteins of each NA strain examined, while reactivities with the glycoproteins of SA strains were substantially weaker and variable and dependent upon both the immune response of the vaccinee and the virus isolate assayed. Most striking was the modest to virtual lack of reactivity with the E2 protein of SA strains. Antigenic differences among the glycoproteins of EEE viruses were not as pronounced in immunoprecipitation analysis. Most significantly, although human immune sera displayed high neutralizing titers against each of the NA isolates examined, only negligible neutralizing titers were obtained against SA isolates. These data suggest that immunized individuals would mount an effective antibody response against infection with NA strains of EEE virus, but that further investigation is clearly warranted to fully assess the protective capability of the vaccine against infection with SA strains. PMID- 7485721 TI - Power, professionalism, and patient advocacy. PMID- 7485722 TI - Hayes Martin Lecture. Training the head and neck surgeon-scientist. PMID- 7485720 TI - Treatment of porcine cysticercosis with albendazole. AB - In a randomized, controlled study, the efficacy and safety of two different schemes of albendazole therapy for treatment of porcine cysticercosis were tested. Seventeen naturally infected pigs were divided into three groups and treated per os with albendazole (50 mg/kg single dose), albendazole (30 mg/kg every day for three days), or given no treatment, respectively. Serologic responses were monitored with the enzyme-linked electroimmunotransfer blot assay. Pigs were humanely killed 12 weeks after treatment, necropsied, and the number of parasites was recorded. Scolex evagination was used to assess viability of the cysts. Both albendazole-treated groups had significant side effects (anorexia, lethargy). Only a single viable cyst was recovered from the brain of one animal after therapy in the multiple-dose group, and the single-dose therapy left 11% of the cysts viable. In contrast, more than 90% of muscle cysts were found to be viable in the untreated group. Although albendazole therapy for three days was found to be highly effective, side effects and the need for multiple doses would still prevent its widespread use. PMID- 7485723 TI - Enhanced tumor cell expression of tumor necrosis factor receptors in head and neck squamous cell carcinoma. AB - BACKGROUND: To determine if tumor necrosis factor (TNF) receptors are upregulated in tumor cells, we measured the distribution and levels of TNF-alpha and TNF beta, and TNF receptors RI and RII, in head and neck squamous cell carcinoma (HNSCC) tumor specimens and normal control specimens. METHODS: HNSCC and control tissue specimens were analyzed qualitatively using immunohistochemistry and quantitatively using immunoassays. RESULTS: Immunohistochemical analysis revealed that TNF-alpha, TNF-beta, TNF RI, and TNF RII antigens were associated predominately with tumor cells in the tissue. Quantitative analysis of TNF factors and receptors in tissue homogenates (mean levels +/- standard error of the mean, in pg/mg of total protein) indicated that: (1) TNF-alpha levels in cancer patients were not statistically different from levels in normal tissues (7.27 +/- 0.91 versus 4.62 +/- 1.33, respectively, P < 0.11); (2) TNF-beta levels in cancer patients were one third of those in normal tissue (5.07 +/- 1.83 versus 16.06 +/- 3.26, respectively, P < 0.01); and (3) both TNF RI and TNF RII levels were consistently elevated two- to four-fold in the cancer tissue when compared to normal tissue levels (1,228.72 +/- 125.67 versus 650.33 +/- 187.70, P < 0.01; and 823.39 +/- 95.90 versus 230.03 +/- 153.01, P < 0.002, respectively). CONCLUSIONS: In HNSCC, enhanced expression of TNF receptors on the cancer cells occurs and is likely to contribute to the regulation of TNF and its activation of tumor cells within the tumor microenvironment; targeting these receptors in cancer cells may provide a new approach to controlling tumor growth and metastasis. PMID- 7485724 TI - Detection of squamous neoplasia by fluorescence imaging comparing porfimer sodium fluorescence to tissue autofluorescence in the hamster cheek-pouch model. AB - BACKGROUND: Early neoplastic changes in the oral cavity may be difficult to detect. Fluorescence imaging using porphyrin-derived drugs has been used to enhance detection of neoplasia. Autofluorescence has also been used for this purpose. This paper compares autofluorescence to porfimer sodium-induced fluorescence in the detection of neoplasia in the hamster cheek-pouch model. MATERIALS AND METHODS: Neoplasia was induced in the hamster cheek pouch by the application of 9,10-dimethyl-1,2-benzanthracene. Animals were imaged either with injection of drug (porfimer sodium) or without drug (autofluorescence). Imaging was carried out using a laser-induced fluorescence detection system. Biopsies were performed on imaged sections and histologic grades were assigned. RESULTS: Porfimer sodium fluorescence provided 100% sensitivity and specificity in detection of neoplasia. The sensitivity and specificity with autofluorescence was 76% and 83%, respectively. CONCLUSIONS: Autofluorescence provides an accurate means of detecting early neoplastic changes in the hamster cheek-pouch model; however, porfimer sodium imaging does improve detection rates. PMID- 7485725 TI - Impact of clinicopathologic parameters on patient survival in carcinoma of the cervical esophagus. AB - BACKGROUND: The survival of patients with carcinoma of the cervical esophagus remains poor in spite of multimodality treatment and technical improvements in surgical resection and reconstruction. This study was undertaken to update our experience with cervical esophageal carcinoma and to identify factors that had an impact on patient survival and quality of life. PATIENTS AND METHODS: Clinical data encompassing 132 variables were collected on 67 patients with cervical esophageal carcinoma from 1980 to 1993. Statistical analysis was performed: independent Student's t-tests, Cox regression, Kaplan-Meier curves, and log rank analyses were used in the statistical evaluation. The mean age of the patients was 63 years (range 31 to 88). Dysphagia was the primary symptom in 86% of patients; 80% had received no prior treatment. The most common abnormal finding (21%) on physical examination was a neck mass. RESULTS: Curative resection was performed in 22 patients, 7 had palliative procedures, and 7 were found to be inoperable at exploration and received palliative treatment. Radiation with or without chemotherapy was definitive treatment for 10 patients, whereas 4 patients were treated with chemotherapy alone for cure, and 17 patients received palliative treatment. The mean survival following diagnosis was 17 months (range 1 to 96). Cumulative 5-year survival was 12%. CONCLUSIONS: Persistent disease, chemotherapy prior to presentation, and chemotherapy for cure remained as statistically significant parameters associated with decreased survival by multivariate analysis. There was a trend toward improved survival in patients treated with surgical resection. PMID- 7485726 TI - Are the new staging rules better for T4 oral cancer? AB - BACKGROUND: In order to find whether the changes in primary tumor definition influence the distribution of T (tumor extent) stages and prognoses, a retrospective study was undertaken. PATIENTS AND METHODS: There were 1,040 patients with adequate data for staging and 868 of those patients underwent surgical treatment. Among the surgically treated patients, 187 fulfilled the new (American Joint Committee on Cancer [AJCC]/International Union Against Cancer 1988) and 77 fulfilled the old (AJCC 1977) criteria for T4 tumors. The distribution of all patients, rate of patients initially treated with surgery, incidence of metastases, and survival were analyzed. RESULTS: The neck was clinically positive (N+) in 50% of T4 patients staged according to the new rules and 69% of patients staged according to the old rules. The 5-year overall survival rate for T4-staged patients irrespective of therapy was 29% using the new rules and 13% using the old rules. The 5-year disease-free survival rate for T4-operated patients was 60% using the new rules and 45% using the old rules. CONCLUSIONS: The previous AJCC rules for the T4 tumors were a better indicator of poor prognosis. PMID- 7485727 TI - Elective neck dissection in the treatment of T3/T4 N0 squamous cell carcinoma of the larynx. AB - BACKGROUND: This study analyzed pathologic findings of clinically occult cervical lymph nodes of T3/T4 N0 squamous cell laryngeal carcinoma and their impact on locoregional failures and overall survival. PATIENTS AND METHODS: A retrospective analysis of 76 patients with T3/T4 N0 laryngeal carcinoma was carried out between 1981 and 1989. Sixty-seven patients had transglottic tumor, 31 patients had extralaryngeal spread, 56 patients were T3 N0, and 20 patients were T4 N0. Seventy-five patients had total laryngectomy and 1 had near total laryngectomy. All patients had bilateral elective neck dissection. The chi-square test was applied to factors related to neck metastasis and locoregional failure. Survival was analyzed using the Kaplan-Meier actuarial method; differences were tested using the Wilcoxon signed-rank test. RESULTS: Eighteen patients had positive surgical margins. Occult neck metastasis was observed in 30%. Univariate analysis showed that cancer stage and cartilage status were not significant to predict neck metastasis. Locoregional recurrence was observed in 28% of patients. Surgical margins, cervical metastasis, lesion extension, and cartilage invasion had significant impact on disease-free survival. The 5-year overall survival was 52%; disease-free survival was 57%. CONCLUSION: The elective bilateral neck dissection performed in T3/T4 N0 patients yielded a 30% incidence of occult neck metastasis. Classification of transglottic carcinomas into endolaryngeal and exolaryngeal provides a better parameter for predicting neck metastasis than does T status. Disease-free and overall survival were significantly affected by neck metastasis, T stage, exolaryngeal tumor, cartilage infiltration, and surgical margins. PMID- 7485728 TI - Factors influencing contralateral metastasis and prognosis from pyriform sinus carcinoma. AB - BACKGROUND: The presence of neck metastasis reduces the probability of regional control and survival. The purpose of this study is to identify risk factors for contralateral neck metastasis in a series of 218 patients with pyriform sinus carcinoma. PATIENTS AND METHODS: Tumor stages were: 31 T1-T2, 187 T3-T4, 40 N0, 174 N1-N3, and 4 NX. A total of 182 patients (83%) had metastatic lymph nodes (23 in the contralateral neck). RESULTS: Thirty-five patients presented neck recurrences (27 in contralateral undissected neck). Logistic regression identified T and N stages, epilarynx and posterior pharyngeal wall involvement, hemilarynx fixation, and ipsilateral level 1 metastasis as important predictors of contralateral metastasis. Five-year survival rates were 41% (pN0), 23% (contralateral pN0), and 31% (contralateral positive nodes). CONCLUSIONS: The contralateral side of the neck was the most common site of recurrence. A lateral neck dissection is advisable for the contralateral side of the neck for high-risk patients. PMID- 7485729 TI - Parotid and neck metastases from cutaneous squamous cell carcinoma of the head and neck. AB - BACKGROUND: Approximately 5% of cutaneous squamous cell carcinomas (SCC) metastasize to regional nodes. Nodal involvement may be more common for cutaneous squamous cell carcinomas of the head and neck, but few large series have reported clinical outcome after treatment of parotid gland and neck metastases from these cancers. PATIENTS AND METHODS: A combined retrospective/prospective study of patients treated between 1983 and 1994 was performed. There were 69 men and 6 women, with a median age of 67 years. Twenty-nine patients had neck metastases, and 33 had parotid gland metastases, while 13 patients had involvement at both sites. Of the 75 patients, 68 were treated surgically and 50 received postoperative radiotherapy. RESULTS: The facial nerve was sacrificed totally in 6 patients and partially in 9. Histologic extranodal spread was present in 48 (71%) of all surgically treated patients. Among 61 patients followed up to recurrence, or for greater than 12 months, 26 (43%) developed recurrence--12 in the parotid gland, 7 in the neck, and 7 in both sites. Multiple recurrences were common and occurred at a median of 8 months after surgery. Positive surgical margins were associated with poorer local disease control (P < 0.05). Cumulative survival at 5 years was 61%, but only 15 of 70 evaluable patients (21%) were eligible for follow-up at this time. Neck involvement with or without parotid gland disease was associated with an increased risk of distant metastases, but this was not statistically significant. Postoperative radiotherapy was not associated with improved disease control. CONCLUSION: Cutaneous, metastatic SCC involving the parotid gland and neck is an aggressive disease with a tendency to an infiltrative growth pattern and multiple recurrences. More aggressive surgery may be justified to reduce the incidence of regional failure after parotidectomy and neck dissection. PMID- 7485730 TI - Prospective study using skin staplers in head and neck surgery. AB - BACKGROUND: A prospective trial was undertaken to investigate the advantages and disadvantages of stapled skin closure versus conventional nylon sutures in head and neck surgery. PATIENTS AND METHODS: The study included 20 consecutive patients who underwent extensive surgery in which their skin was closed with staples. Another group of 20 matched patients receiving a noncontinuous nylon suture closure was followed in parallel. RESULTS: The complications recorded occurred in 5 patients in the stapled group and 3 in the sutured group. Analysis of cosmetic results showed 16 patients (80%) in the stapled group with good wound appearance and 17 (85%) in the sutured group. The mean closure time was 5 minutes for the stapled group and 25 minutes for the sutured group. Cost was $19.75 for conventional closure and $22.00 for mechanical suture. CONCLUSION: The use of skin staples speeds up closure time by 80%, yields similar cosmetic results with no increase in complications, although at a slightly higher cost. PMID- 7485731 TI - Surgical rescue of recurrent carcinoma of the nasal columella. AB - BACKGROUND: Tumors involving the nasal columella have been noted for their aggressive behavior. Columellar malignancies exhibit perhaps the highest incidence of recurrence and metastasis of all nasal and midfacial cancers of cutaneous origin. Periosteal invasion of maxillary bone and nasal floor may occur "barrier free." Submucosal infiltration of the nasal septum easily develops, leading to potentially lethal invasion of the anterior cranial base. Rapid extension of cancer to the adjacent bony and cartilaginous facial structures requires extensive oncological resections for control. While tumor histology and biologic behavior remain important prognostic indicators, anatomic location within the columella carries with it a much poorer prognosis than do other nasal cutaneous lesions. Recurrent tumors in this location as a result of inadequate initial treatment present a formidable therapeutic challenge. PATIENTS AND METHODS: Fourteen patients who had initially been "adequately" treated for cutaneous malignancies of the nasal columella were seen from 1982 to 1992 by the authors because of local tumor recurrence. Histologically, 4 of the 14 were initially basal cell epithelioma, 7 were squamous cell epithelioma, 1 was squamous/basal cell epithelioma, 1 was melanoma, and 1 was anaplastic. By the time tumor regrowth was recognized (mean 24 months), 100% demonstrated bony involvement of the premaxilla or nasal floor, 57% (8 of 14) nodal metastasis to the parotid or neck, and 43% (6 of 14) extension to the anterior skull base. Thirty-six percent (5 of 14) had undergone unsuccessful radiation therapy in an attempt to control the disease. RESULTS: All patients underwent radical oncological surgery with or without postoperative radiation therapy in an attempt to control the tumors, including combinations of anterior skull base resection, partial or total rhinectomy, partial maxillectomy, and parotidectomy with radical neck dissection. At 5 years, 5 patients (36%) had no evidence of disease and 1 (7%) was alive with disease; at 54 months, 6 (43%) had no evidence of disease and 1 (7%) was alive with disease; at 42 months, 7 (50%) were alive with no disease. CONCLUSIONS: In this series, an aggressive surgical approach successfully salvaged 43% of recurrences at 5 years and 50% at 54 months. Although overall prognosis remains poor, radical surgery appears a viable modality for rescue of recurrent malignancies of the columella. In view of the ability to control recurrence, more aggressive initial therapy might improve the prognosis of the early columellar lesion. PMID- 7485732 TI - Treating tumors of the sublingual glands, including a useful technique for repair of the floor of the mouth after resection. AB - BACKGROUND: From a 55-year experience with about 4,000 patients treated for salivary gland tumors, we have identified 18 patients (0.5%) who received definitive treatment for neoplasms arising in the sublingual glands. PATIENTS AND METHODS: Patient ages ranged from 35 to 76 years (median 59), and men and women were equally represented. All had malignant tumors, which included adenoid cystic carcinoma (9 patients), mucoepidermoid carcinoma (5 patients), and adenocarcinoma (4 patients). An asymptomatic swelling was the most common complaint (7 patients); the tumor was incidentally discovered by a dentist in 4 others. For all patients, the treatment delay ranged from 3 to 24 months, but only 3 had lesions that extended beyond the confines of the floor of the mouth. All were treated surgically by resections, which were peroral in 4 patients, transcervical in 5, and pull-through type (including the submandibular gland) in 7. The remaining 2 patients had composite procedures. In 3 more recently treated patients, pull-through resections were followed by a floor-of-the-mouth repair involving transmandibular sutures. This technique has proved useful and will be described in detail. RESULTS: Results in this small patient cohort have been most encouraging. Local recurrence occurred in 3 patients who succumbed 11 months, 27 months, and 21 years after initial treatment. Fourteen others remain alive and well 20 months to 20 years (median 74 months) posttreatment, and the remaining patient was recurrence free when he died of unrelated causes 7 years after resection. CONCLUSION: This small experience confirms the rarity of sublingual gland tumors and attests to the good results in terms of function and survival that can be anticipated with adequate surgical treatment when the lesion is relatively small. PMID- 7485734 TI - Differentiated carcinoma of the thyroid with extrathyroidal extension. AB - BACKGROUND: We have analyzed our experience with differentiated thyroid cancer patients with extrathyroidal extension (ETE) to investigate patterns of recurrence and define factors that predict failure. PATIENTS AND METHODS: The records of 1,012 patients treated surgically from 1930 to 1985 were reviewed. A total of 79 patients (8%) had ETE. The median length of follow-up was 10 years. RESULTS: Patients with ETE were more likely to fail treatment and to die of their disease than were patients without ETE (77% versus 34% and 71% versus 13%, respectively; P < 0.0001). Local, regional, and distant failures were more prominent among patients with ETE than among those without ETE (48% versus 9%, 41% versus 16%, and 37% versus 11% respectively; P < 0.0001). Survival of patients with ETE was adversely affected by nonpapillary histology, distant metastasis, age > 45, tumor size > 4 cm, and incomplete excision (P < or = 0.05). After stratification for age, survival in older patients was not affected by tumor size or incomplete excision, while in younger patients tumor size or the presence of distant metastasis did not adversely affect survival. Patients younger than 45 with negative margins had similar survival to patients without ETE (P = 0.46). CONCLUSIONS: Patients with ETE are more likely to die of their disease and to fail at all sites. Survival in older patients was not affected by incomplete excision while it was in younger patients. The presence of distant metastasis did not affect survival in younger patients. Our results suggest that among patients under 45, the presence of ETE does not adversely impact upon survival when the primary tumor is completely resected. PMID- 7485733 TI - Prediction of potential metastatic sites in cutaneous head and neck melanoma using lymphoscintigraphy. AB - BACKGROUND: The technique of lymphoscintigraphy may allow a more selective approach to the management of clinically negative neck nodes among patients with cutaneous head and neck melanoma. PATIENTS AND METHODS: A group of 97 patients with cutaneous head and neck melanoma had preoperative lymphoscintigraphy using intradermal injections of technetium 99m antimony trisulfide colloid to identify sentinel nodes. Fifty-one patients were eligible for clinical analysis after initial definitive treatment by wide excision only (n = 11), wide excision and elective dissection of the neck (n = 19) or axilla (n = 1), or wide excision and a sentinel node biopsy procedure (n = 20). RESULTS: Sentinel nodes were identified in 95 of 97 lymphoscintigrams, and 85% of patients had multiple sentinel nodes. In 21 patients (22%), sentinel nodes were identified outside the parotid region and the 5 main neck levels, mostly in postauricular nodes (n = 13). Lymphoscintigrams were discordant with clinical predictions in 33 patients (34%). Lymph nodes were positive in 4 elective dissections and 4 sentinel node biopsies. Among 16 patients evaluable after wide excision and a negative sentinel node biopsy, 4 patients subsequently developed metastatic nodes; however, confident identification of all nodes marked as sentinel nodes on lymphoscintigraphy was not achieved at the original biopsy procedure in 3 of these patients. CONCLUSIONS: Lymphoscintigraphy and sentinel node biopsy are more difficult to perform in the head and neck than in other parts of the body. The reliability of sentinel node biopsy based on lymphoscintigraphy may be improved by identifying and marking all nodes that are considered to receive direct lymphatic drainage from the primary melanoma, and by use of a gamma probe intraoperatively. PMID- 7485735 TI - Age correlates with increased frequency of high risk factors in elderly patients with thyroid cancer. AB - BACKGROUND: Age and certain pathologic features (ie, follicular histology, vascular invasion, and extracapsular extension) are reported to impact on prognosis in thyroid cancer, but the relationship between these two factors remains incompletely defined. PATIENTS AND METHODS: Clinical, pathologic, therapeutic, prognostic, and outcome data were retrospectively collected and compared among three groups of patients with thyroid cancer: group 1 (age 21 to 50), group 2 (age 51 to 70), and group 3 (age greater than 70). RESULTS: There was a statistically significant age-related decrement in survival: 10-year survival was 92% in group 1, 77% in group 2, and 48% in group 3. Follicular histology, vascular invasion, and extracapsular extension were more frequently observed in group 3 versus group 2 and in group 2 versus group 1 (all differences statistically significant except for vascular invasion). Although survival was significantly less for group 3 versus group 2 versus group 1, only vascular invasion and extracapsular extension independently predicted outcome by multivariate analysis. CONCLUSIONS: The prognostic importance of age in thyroid cancer may be due to the greater prevalence of pathologic risk factors in older patients. PMID- 7485736 TI - An evaluation of prognostic scoring systems in patients with follicular thyroid cancer. AB - BACKGROUND: Prognostic scoring systems for thyroid cancer have not been investigated in patients with pure follicular cancer; thus, the purpose of this study was to compare the following prognostic indices: the European Organization for Research and Treatment of Cancer (EORTC) method; the Age, Grade, Extent, Size (AGES) score; and the Age, Metastasis, Extent, Size (ALIES) score. PATIENTS AND METHODS: A retrospective study reviewing 122 patients actively treated between 1955 and 1990 was conducted. Scoring systems were calculated and survival analysis completed. AGES low-risk patients were analyzed with respect to known risk factors. RESULTS: The AGES scoring system significantly defined low- and high-risk groups (P = 0.0041); the ratio of deaths between high-versus low-risk groups was 1.9:1. EORTC scores distinguished four risk groups (P = 0.002). The AMES scoring system did not significantly assign risk. In multivariate analysis of low-risk AGES patients, age, perithyroidal tissue involvement, and positive frozen section reached statistical significance. CONCLUSIONS: The AGES and EORTC scoring systems best defined low- and high-risk groups of patients with pure follicular cancer, although the separation between groups was low. PMID- 7485737 TI - Management of the hot thyroid nodule. AB - BACKGROUND: Solitary hyperfunctioning nodules of the thyroid gland are usually viewed as benign. They may present with autonomous euthyroidism but are of concern for potential progression to hyperthyroidism. Various methods of treatment are worthy of consideration. PATIENTS AND METHODS: Forty-five patients with solitary hot thyroid nodules verified by radioisotope scintiscanning were selected for treatment. Thirty-one underwent surgery, usually partial thyroidectomy. Eight euthyroid patients received no treatment, 5 underwent therapy with radioactive iodine (RAI), and 1 received thyroid suppression treatment. The cases were assessed retrospectively. RESULTS: Thyroidectomy patients had no morbidity, were well, and showed 1 Hurthle cell tumor and 5 coincidental small malignancies associated with benign hot nodules, including a contralateral cancer. Untreated patients showed continuance of good health, but nodules persisted and 1 Graves' orbititis occurred. The RAI-treated patients had persistent nodularity, improved function, and 1 case of hyperparathyroidism. Thyroid feeding only caused iatrogenic toxicity and was discontinued. CONCLUSIONS: There are various techniques for managing the hot nodule. Nonsurgical methods may be effective, but can result in persistent nodularity and iatrogenic sequelae. Excision had no morbidity in this series and was effective in providing immediate relief of problems present and potential. PMID- 7485738 TI - Investigation of calcium-induced hydrolysis of phosphoinositides in normal and lithium-treated parathyroid cells. AB - BACKGROUND: Lithium-induced hyperparathyroidism is characterized by a reduction in parathyroid sensitivity to changes in extracellular calcium (Ca2+). Ca(2+) induced transmembrane signal transduction in the parathyroid cell is known to result in the hydrolysis of phosphatidylinositol bisphosphate (PIP2), generating increases in intracellular inositol phosphates, a process which is mediated by a calcium receptor. MATERIALS AND METHODS: To determine if lithium's effect on parathyroid cell function is mediated by an alteration in Ca(2+)-induced hydrolysis of PIP2, inositol 4-monophosphate (IP1), and inositol 1,4,5 trisphosphate (IP3) were measured using anion-exchange chromatography in normal and lithium chloride (LiCl)-treated bovine parathyroid cells at Ca2+ concentrations varying from 0.5 mmol/L to 5.0 mmol/L. IP1 and IP3 concentrations were determined in terms of percent control, defined as the IP1 or IP3 concentration at an [Ca2+] of 0.5 mmol/L. RESULTS: Increases in [IP1]/10(6) cells (mean +/- standard error of the mean [SEM]) in response to progressive increases in Ca2+ from 0.5 mmol/L to 5.0 mmol/L varied from 825 +/- 228 to 4,474 +/- 382 in control cells versus 1,139 +/- 243 to 4,689 +/- 630 in cells pretreated with LiCl (P > 0.05). The increases in [IP3]/10(6) cells (mean +/- SEM) in response to increases in Ca2+ from 0.5 mmol/L to 5.0 mmol/L, varied from 146 +/- 14 to 385 +/ 35 in control cells versus 134 +/- 16 to 327 +/- 55 in cells pretreated with LiCl (P > 0.05). CONCLUSIONS: Our results demonstrate that LiCl does not effect Ca(2+)-induced hydrolysis of PIP2, suggesting that the desensitizing effect of LiCl on the parathyroid cell is not the result of a Ca2+ receptor-mediated phenomenon. PMID- 7485739 TI - Analysis of savings in operative time for primary hyperparathyroidism using localization with technetium 99m sestamibi scan. AB - BACKGROUND: The cost effectiveness of preoperative localization in cases of primary hyperpara-thyroidism has not been established. We analyzed the potential savings in operative time after localization with technetium 99m (99mTc) sestamibi scan. METHODS: Thirty-three patients had localization of a solitary adenoma with 99mTc-sestamibi. Measurement was made of the time required for adenomectomy, unilateral neck exploration (UNE), unilateral neck exploration and confirmation of one contralateral parathyroid gland (UNEC), or bilateral neck exploration (BNE). RESULTS: The total operative time in minutes was 76.4 +/- 18.8 for adenomectomy; 87.5 +/- 20.4 for UNE; 105.6 +/- 25.0 for UNEC; and 117.9 +/- 26.7 for BNE. The time difference was significant between adenomectomy versus UNE, UNEC, and BNE. There were also significant time differences between UNE versus UNEC and BNE. CONCLUSIONS: The preoperative localization of a solitary parathyroid adenoma may optimize operative time with UNE, saving approximately 30 minutes. PMID- 7485740 TI - TP53 mutations in upper aerodigestive squamous cell carcinomas from a group of Brazilian patients. AB - BACKGROUND: Loss of function of the tumor suppressor gene TP53 contributes to the development of several tumors. PATIENTS AND METHODS: We screened DNA samples from 47 patients with upper respiratory system squamous cell carcinomas for the presence of TP53 mutations. Exons 4 to 8 of the TP53 gene were amplified by the polymerase chain reaction, and mutations were identified by single-strand conformation polymorphism analysis. RESULTS: The TP53 mutations were demonstrated in 23 cases (49%). Mutations were distributed as follows: exon 4, 5 cases; exon 5, 4 cases; exon 6, 6 cases; exon 7, 4 cases; and exon 8, 4 cases. Demographic variables, tumor site, stage, family history of cancer, and tobacco smoking were not predictors of TP53 mutations. There was an increasing number of mutations in the more undifferentiated tumors (P = 0.0594). CONCLUSIONS: These findings suggest that TP53 mutations are associated with tumor differentiation, but not with the risk of lymph node metastasis in the group of patients analyzed. PMID- 7485741 TI - Native cellular fluorescence can identify changes in epithelial thickness in-vivo in the upper aerodigestive tract. AB - BACKGROUND: Change in epithelial thickness is part of the neoplastic transformation process of the upper aerodigestive tract. The quantitation of native cellular fluorescence (NCF) may represent a noninvasive means of distinguishing such a change. PATIENTS AND METHODS: Nineteen patients with squamous neoplasms and 12 surgical specimens from cancer patients were analyzed for NCF using a hand-held fiber optic probe attached to a fluorescent spectrometer. Tumors and normal sites were analyzed for fluorescence, and tissue samples were obtained. Ratios of intensities of various emitted wavelengths were computed to quantitate and compare various spectral patterns. These ratios were then correlated with mucosal thickness. RESULTS: The 330 nm peak in the excitation scan (lambda Ex 200 to 360 nm, lambda Em 380 nm) was lost in the tumors compared with the normal sites. The 390 nm peak in the emission scan (lambda Ex 340 nm, lambda Em 360 to 660 nm) was also lost. The 290 nm/330 nm ratio in the in-vivo excitation scan (lambda Ex 200 to 360 nm, lambda Em 380 nm) correlated with changes in epithelial thickness. The 390/450 ratio in the emission scan (lambda Ex 340 nm, lambda Em 360 to 660 nm) correlated negatively with the mean epithelial thickness. CONCLUSIONS: Native cellular fluorescence analysis can identify changes in neoplastic tissues, including changes in epithelial thickness. PMID- 7485742 TI - Glutathione s-transferase genotypes as risk factors for head and neck cancer. AB - BACKGROUND: Several enzymatic systems, including glutathione S-transferases, are involved in the metabolism of environmental agents. The absence of glutathione S transferases mu (GSTM1) and theta (GSTT1) results in decreased detoxification of carcinogens, for example, chemicals in cigarette smoke. These metabolic deficiencies may predispose individuals to the development of smoking-related tumors, such as cancers of the lung, head and neck, and bladder. METHODS: The glutathione S-transferase genotypes of 186 previously untreated patients with squamous cell carcinoma of the head and neck and 42 healthy controls were determined with polymerase chain reaction (PCR) methodologies. Lymphocytes separated from heparinized peripheral blood or whole blood extracts served as sources of genomic DNA. The presence or absence of the gene-specific PCR products revealed the positive or negative genotypes, respectively. RESULTS: The absence of the GSTM1 genotype conferred an odds ratio of 2.37, and the 95% confidence interval (CI) was 1.20 to 4.67. The absence of the GSTT1 gene conferred an odds ratio of 1.47 (CI 0.71 to 3.02). In the population of 42 patients and their matched 42 controls, the absence of the GSTM1 and GSTT1 genotypes conferred odds ratios of 3.10 (CI 1.24 to 7.75) and 2.18 (CI 0.91 to 5.23), respectively. CONCLUSIONS: Despite the small study size, our preliminary data suggest that genetically determined factors of carcinogen metabolism may be associated with increased risk for head and neck cancer. PMID- 7485743 TI - The effect of thrombospondin on oral squamous carcinoma cell invasion of collagen. AB - BACKGROUND: Thrombospondin (TSP), a cell matrix protein, and transforming growth factor beta (TGF-beta), a growth regulatory protein, play roles in tumor progression. The purpose of this study was to investigate the effects of TSP and TGF-beta on tumor cell invasion. MATERIALS AND METHODS: Tumor cell invasion assays were performed using a modified Boyden chamber apparatus with collagen coated membranes. The KB oral carcinoma cell line was studied in serum-free media. Invasion was measured as the summation of the number of cells in five representative low-power fields (x 100) traversing the collagen barrier after a 3 hour incubation period. The effects of antibodies against TSP, TGF-beta and the cysteine-serine-valine-threonine-cysteine-glycine (CSVTCG)-specific TSP receptor were also evaluated. RESULTS: TSP caused a dose-dependent stimulation of tumor cell invasion. Antibodies against TSP, its CSVTCG-specific receptor, and TGF-beta inhibited TSP-promoted invasion by 50% to 71%. CONCLUSIONS: TSP and its CSVTCG specific receptor promote KB cell invasion of collagen through the production and/or activation of TGF-beta. PMID- 7485744 TI - Improved survival with preoperative chemotherapy followed by resection uncompromised by tumor response for advanced squamous cell carcinoma of the head and neck. AB - BACKGROUND: A total of 93 patients were treated with one of two preoperative chemotherapy regimens over a 15-year period. The study supports the importance of strict adherence to guidelines for ablative surgery. METHODS: A single surgeon performed the surgery and evaluated each patient prior to treatment. The extent of the planned operation was documented. RESULTS: The 5-year absolute survival of 88 patients who completed the protocol was estimated at 55%. The 40 cisplatin/5 fluorouracil-treated patients exhibited a significantly better outcome than the 48 cisplatin/bleomycin-treated patients (76% versus 43%, respectively). Comparison of a subset of 37 patients with a matched group from the standard control arm of the Head and Neck Contracts Program demonstrated a statistically significant improvement in overall survival over standard treatment. CONCLUSIONS: These data suggest that strict adherence to preoperative chemotherapy planning of ablative uncompromised surgery contribute to improved survival. Selective rather than routine postoperative radiotherapy may be advantageous. PMID- 7485746 TI - Long-term evaluation of patients undergoing immediate mandibular reconstruction. AB - BACKGROUND: Immediate reconstruction following segmental mandibulectomy is an accepted surgical technique. The benefits and patient selection criteria need to be established. PATIENTS AND METHODS: Forty-seven patients who underwent immediate reconstruction of the mandible were followed for up to 14 years. Survival, complication rates, and functional results were analyzed. RESULTS: Median survival was 39 months and actuarial 5-year survival was 40%. Two patients died in the postoperative period, and 9 suffered major reconstruction-related complications. The majority of these complications were related to the use of reconstruction plates, and occurred when the mandibular defect included the arch or when the plates were covered by pectoralis flaps. Half of the patients interviewed were able to eat a regular diet. CONCLUSIONS: Mandibular reconstruction can be performed safely and expeditiously in nearly all patients undergoing segmental mandibulectomy. Functional results and long-term survival will be acceptable in many cases. PMID- 7485748 TI - Congenital cervical bands in thoracic outlet syndrome. PMID- 7485745 TI - Preoperative cisplatin and accelerated hyperfractionated radiation induces high tumor response and control rates in patients with advanced head and neck cancer. AB - BACKGROUND: Preoperative therapy with concurrent cisplatin infusion and accelerated hyperfractionated radiation has the potential to eradicate pools of resistant cells with high-growth fraction that accumulate during standard radiation therapy. Based on preliminary data showing a high complete response rate (necessary for organ preservation) in patients with advanced squamous cell cancer of the head and neck (stages III and IV), we conducted a multi institutional study using accelerated hyperfractionated radiation to maximize local and regional tumor control without adding toxicity. PATIENTS AND METHODS: Preoperative radiation was given in 1.8 Gy fractions for 2 weeks followed by 1.2 Gy BID hyperfractionation to 46.7 Gy. Continuous infusion of cisplatin 20 mg/m2 was given on days 1 through 4 and 22 through 25. Patients were initially assessed with panendoscopy and were subsequently reassessed with biopsy of primary site (+/- nodes). Patients with negative primary site biopsy received additional hyperfractionated radiation to 76 Gy with simultaneous carboplatin 25 mg/m2 BID for 12 consecutive days. Patients with residual disease after 47 Gy had resection of primary site and node dissection. Of 69 patients treated, 56 were T3 and T4, and 46 were N1, N2, and N3; 17 patients (25%) were stage III and 50 (72%) were stage IV; in 2 patients (3%) the stage was unknown. RESULTS: Treatment was well tolerated with no grade IV toxicities, but there were 13 patients with grade III toxicities (mucositis, dysphagia, or hematological toxicity). Biopsy specimens were taken from 84% of the patients, and 81% of the primary sites showed pathologic complete response and 49% of the lymph nodes showed a pathologic complete response. Thirty-four patients underwent curative surgery. Eleven required resection of the primary and 23 required neck dissection only. At 2 years (median followup 12 months), 44 of 69 patients (64%) have no evidence of disease, 9 are dead of disease, 9 are dead of other causes, and 6 are alive with disease. Surgical toxicities were minimal and primarily limited to fibrosis in 12 patients, edema in 8 patients, tracheoesophageal fistula in 1 patient, and delayed wound healing in 1 patient. Late toxicities included xerostomia in 22 patients, dysphagia in 17, and speech impairment in 9. CONCLUSIONS: Preoperative concurrent radiotherapy and chemotherapy provides a high level of organ preservation and local and regional control because of the high complete response rate at the primary site. A planned neck dissection with minimal morbidity ensures local/regional control, as well as providing definitive staging information. A randomized evaluation is suggested for this program. PMID- 7485747 TI - Clinical experience with a microvascular anastomotic device in head and neck reconstruction. AB - BACKGROUND: Despite numerous refinements in microsurgical technique and instrumentation, the microvascular anastomosis remains one of the most technically sensitive aspects of free-tissue transfer reconstructions. MATERIALS AND METHODS: Concurrent with the development of microsurgical techniques, various anastomotic coupling systems have been introduced in an effort to facilitate the performance and reliability of microvascular anastomoses. The microvascular anastomotic coupling device (MACD) studied here is a high-density, polyethylene ring-stainless steel pin system that has been found to be highly effective in laboratory animal studies. Despite its availability for human clinical use over the last 5 years, reported clinical series remain rare. Our clinical experience with this MACD in 29 head and neck free-tissue transfers is reported herein. RESULTS: Thirty-five of 37 (95%) attempted anastomoses were completed with 100% flap survival with a variety of donor flaps, recipient vessels, and clinical contexts. Two anastomoses were converted to conventional suture technique intraoperatively, and one late postoperative venous thrombosis occurred after fistulization and vessel exposure. CONCLUSIONS: We conclude that the MACD studied here is best suited for the end-to-end anastomosis of soft, pliable, minimally discrepant vessels. Previous radiation therapy does not appear to be a contraindication to its use. Interpositional vein grafts may also be well suited to anastomosis with the device. When carefully and selectively employed by experienced microvascular surgeons, this MACD can be a safe, fast, and reliable adjunct in head and neck free-tissue transfer reconstructions, greatly facilitating the efficiency and ease of application of these techniques. PMID- 7485750 TI - [Current approaches to the treatment of endometriosis]. PMID- 7485749 TI - Obturator hernia needs an early diagnosis and treatment. PMID- 7485751 TI - [Current approaches to the diagnosis and management of patients with tumors and tumor-like formations of the ovaries]. PMID- 7485752 TI - [Prevention and treatment of hemorrhage in obstetrical hospitals]. AB - Introduction of a program of prophylactic and organizational and therapeutic measures helped reduce the incidence of massive blood loss in 741 women at various modes of delivery, including cases with obstetrical bleedings. The incidence of hemorrhages in the placental and postpartum periods reduced from 1.6 to 0.3%. PMID- 7485753 TI - [Contents of alpha-fetoprotein in the blood of pregnant women as a criterion of the presence of congenital heart defects in the fetus]. AB - alpha-Fetoprotein (AFP) was measured in the blood of 16 women pregnant with twins at various terms of gestation and 24 pregnant women whose fetuses were found to have anencephaly, patent spina bifida, gastroschisis, renal polycystosis, or Down's disease. In Down's disease AFP level was 7 ng/ml (0.17 multiple of medians, MoM) at 17 weeks gestation and 6 ng/ml (0.12 MoM) at 19 weeks. In the fetal abnormalities studied AFP level was 372 ng/ml on average (6.8 MoM) at 16 to 18 weeks gestation, this being about 10 times higher than the normal level. AFP level in twin pregnancy at the same period was 2.3 MoM. AFP measurements are important for the prenatal diagnosis of fetal status in order to plan follow-up of pregnancy and labor management. PMID- 7485754 TI - [Characteristics of adaptive reactions of pregnant women with chronic inflammatory gynecologic diseases and in their newborn in the early neonatal period]. AB - The sympathoadrenal, parasympathetic, and humoral mechanisms of regulation of the autonomic nervous system, as well as the strain and reserve of the adaptive reactions of the body were examined in 57 pregnant women with chronic inflammations of the genitals and their newborns in the early neonatal period. The sympathoadrenal autonomic regulation predominated in pregnant women, whereas in the newborns imperfect adaptive restructuring of the body was observed in the early neonatal period. PMID- 7485756 TI - [Dynamics of the activity of redox enzymes in peripheral blood lymphocytes of the newborn with a history of chronic intrauterine hypoxia]. AB - Study of the cytochemical characteristics of enzymatic activity of peripheral blood cells in the newborns with a history of medium-severe-chronic intrauterine hypoxia revealed shifts in the levels of succinate dehydrogenase and a glycerophosphate dehydrogenase. These data imply that changed enzymatic activity of dehydrogenases is an unfavorable prognostic sign indicating failure of adaptation and deterioration of reactivity in the early neonatal period. PMID- 7485755 TI - [Characteristics of lymphocytic enzyme status of the newborn after metabolic therapy of pregnant women]. AB - The enzymatic synthetic activity of peripheral blood lymphocytes was changed in the newborns whose mothers suffered from habitual miscarriages and were administered courses of metabolic therapy as part of combined pathogenetic treatment (basis therapy). Metabolic therapy brought about positive shifts which manifested by a reliable activation of succinate dehydrogenase and alpha glycerophosphate dehydrogenase, this, in its turn, creating favorable conditions for the functioning of the mother-placenta-fetus metabolic system and being conductive to a better adaptation of the newborns in the early neonatal period. Metabolic therapy brought about a twofold reduction of the incidence of fetal hypoxia and hypotrophy. PMID- 7485757 TI - [Infectious diseases of the female reproductive system]. PMID- 7485758 TI - [Characteristics of hemodynamics in cardiomyopathies of newborn infants of diabetic mothers]. AB - The hemodynamics was studied in 42 newborns with diabetic fetal diseases. Clinical and instrumental examinations revealed cardiomyopathy in 28 of these. Hemodynamic disorders were characterized by a low stroke volume resultant from reduced diastolic volume of the left ventricle. The most grave hemodynamic disorders! were detected in patients with the obstructive cardiomyopathy when both diastolic and systolic volumes of the left ventricle were changed, myocardia contractility reduced, and diastolic function suppressed. PMID- 7485759 TI - [Assessment of the health status of children born as a result of extracorporeal fertilization and artificial insemination]. AB - Comprehensive assessment of the health status of 142 children born In Research Center of Obstetrics, Gynecology, and Perinatology, Russian Academy of Medical Sciences, showed that 92% of children not only survived, but were healthy and their neuropsychic development was normal for age. The hormonal and immunity status was virtually normal. The authors come to a conclusion that extracorporeal fertilization and artificial insemination in the treatment of a sterile marriage are Justified and effective. The health status of the children who are born is normal, provided certain indications are adhered to. PMID- 7485760 TI - [Experience in the treatment of 152 women with bilateral injuries of the ureter]. AB - Data on the treatment of 152 women with bilateral injuries to the ureters are presented. Many-year experience gained in the treatment of women with such injuries permitted the authors to single out their main causes and to find the most rational methods of derivation of the urine and develop the treatment policy depending on the type of injury to the ureters, complications of these injuries, and anatomo-functional status of the urinary system. Plastic surgery brought good results in 84.4% of patients. PMID- 7485761 TI - [Changes in the contents of phosphoinositides in formed elements of blood and endometrial tissue in patients with hyperplastic processes and cancer of the endometrium]. AB - The levels of phosphoinositides were measured in blood macrophages, platelets, lymphocytes, granulocytes, neutrophils, and red cells, as well as in endometrial tissue of 175 patients with glandular hyperplasia of the endometrium, atypical hyperplasia, endometrial polyps and cancer. A reliable reduction of the content of phosphatidyl inositides and phosphatidyl inositide-S-phosphates was observed in all formed elements of the blood, the most marked decrease being observed in endometrial cancer. On the contrary, the content of phosphatidyl inositide-4 phosphates and phosphatidyl inositide-4.5-diphosphates in the macrophages, lymphocytes, and granulocytes was reliably increased in the patients as against healthy women. The process of phosphatidylinositide phosphorylation run an absolutely different course in pathologically altered endometrial tissue than in comparison with that in blood cells. A new phosphatidyl inositide-signal mechanism was revealed, which is unrelated to inositol-1.4.5-triphosphate, as a result of which the potentiality of appearance of new secondary messengers actively participating in cell growth increases. PMID- 7485762 TI - [Epidural anesthesia as a method of labor analgesia preserving motor activity of parturient women]. PMID- 7485763 TI - [Treatment of newborn infants with cephalohematoma]. PMID- 7485764 TI - [Transcatheter embolization in chronic pain syndrome caused by varicose dilatation of ovarian veins]. PMID- 7485765 TI - [Clinical aspects, diagnosis and treatment of intrauterine infections]. PMID- 7485766 TI - [The treatment of urinary stress incontinence with the sling and Pereyra's suspension]. AB - Patients with proven stress incontinence were subjected to surgical treatment which combined both sling and Stamey-Pereyra techniques. A 3-4 cm long and 1,5-2 cm wide sling of Bulgarian polyamide mesh was placed under the urethrovesical junction using a small vaginal incision. Two long polyamide sutures were tied to either end of the mesh and were retrieved from the vagina to the abdomen by Pereyra needle inserted through two small suprapubic incisions and guided by the vaginal forefinger. The technique of the procedure is described. A total number of 27 patients were operated on. Two of them has their suspending sutures cut postoperatively because of prolonged urinary retention. The remaining 25 were continent. The technique proposed combines the advantages of the sling procedures (high efficiency especially in cases of severe and recurrent incontinence) and the simplicity of Stamey-Pereyra suspension technique. On the other hand the Bulgarian polyamide mesh showed very good qualities. No infection, rejection or sling erosion were observed. PMID- 7485767 TI - [The ultrasonic assessment of the cicatrix after a past cesarean section]. AB - A supersonic study of the lower uterine segment of pregnant women which have had a Caesarean operation in the past has been made. The aim of this study is the confirmation of the supersonic scanning as a part of the struggle for diminution of the second abdominal deliveries. Twenty-six pregnant women with anamnesis of Caesarean operation were observed. The following conditions have been kept: precise probable term of delivery, gestational period--35-37 g.w., one foetus in head position, unspoiled amniotic fluid and full urinary bladder. It is obligatory a longitudinal and cross scanning in the area of the cicatrix to be made. For the assessment of the cicatrix sufficiency the author has applied supersonic criteria known in the literature. They include: shape, thickness, incessancy, outside lines and echo structure of the lower uterine segment. The results show: the scar is sufficient in 24 women--it has a triangular form in the lower uterine segment, even outside lines, the thickness of the cicatrix is above 3-5 mm it is incessant and with homogeneous echo structure. In 2 of th pregnant women there are data for inadequacy-the form is like a balloon, the cicatrix has got thinner under 3 mm, the incessancy of the lower segment is absent-it is with defects and uneven outside lines, the echo structure is heterogeneous with predominance of increased echogenicity. According to the way of delivery the results are confirmed pathohistologically, by microscope or by manual palpation. The effectiveness of the supersonic assessment of the cicatrix has been shown. PMID- 7485768 TI - [A single-stage 2-layer suture in cesarean section--the effect of the surgical technic on postoperative febrile conditions]. AB - The aim of the authors is to study the influence of the figure-of-eight suture of the uterine wall on the postoperative febrile complications. 1018 women, delivered with Caesarean were followed. 937 of them with conventional two layer suture of the uterine wall and the rest 81 with a figure-of-eight suture. Postoperative febrile complications were 15.58% in those with two layer suture against 3.5% in the rest 81. Using dispersion analysis the authors point that the on the type of uterine wall suture depends the rate of the postoperative febrile complications. The rate of influence of the figure-of-eight suture is up to 10%. Having in mind that this factor "suture", should be considered as 99% influential with a risk for mistake no more than 1%--F (8.06) > FT (6.64). The results presented show that the figure-of-eight suture is to be preferred to the conventional two layer suture of the uterine wall. PMID- 7485769 TI - [Thromboembolic disease in the puerperium (is thrombotic prophylaxis necessary in the puerperium)]. AB - The author makes retrospectively investigation in 3064 young mothers about determination on dependence between thromboembolism and varices in early puerperium. He finds varices in 1.01% of cases from which 84% are without prophylaxis. In 42% of them with complaints are applied medicine. In 0.13% of cases in puerperium develops thrombophlebitis (0.08-1.35% in bibliography information). The author finds in 0.098% of cases pulmonary thromboembolism (0.04 0.05% in bibliography information). In 77% of them he find out birth traumatism. In conclusion the author recommends prophylaxis with Heparin in early puerperium by cases with previous thromboembolism, traumatic parturition and doubt for infection. PMID- 7485770 TI - [Changes in the weight of the uterus, ovaries and adrenal glands in sexually mature rats treated with estradiol and reserpine]. AB - Immature rats of the Wistar strain were used for the experiments. The treatment was administrated on the 35, 36, 37th day after delivery. The animals were divided into 4 groups. The first group received only saline solution, the second group were treated with reserpine, the third group with estradiole, and the forth group with both reserpine and estradiole. The animals were sacrificed on the 38th day after delivery. There was a statistically significant increase in the weight of the uterus and the ovaries after treatment with estradiole alone, and the same was observed for the group treated with reserpine and estradiole. The latter showed less increase in the uterine weight, which might be due to an impairment of estradiole synthesis caused by reserpine. The wight of the adrenal glands of the immature female rats was increased after treatment with estradiole and reserpine. This implies an involvement of the adrenergic system in the interaction between estradiole and testosterone in immature rats. PMID- 7485772 TI - [Laser therapy as a method for treating condylomata at a prepubescent age]. AB - The Nd:YAG Laser is one of the most effective methods for treatment of acute condylomata. This method has been used at the Children Gynaecological Consulting Room at the Desk of Obstetrics and Gynaecology--University of Medicine,Varna, for two years. During this period seven girls (children) were treated (0.6%) from all the patients treated for external genital inflammation. Only one of the cases needed a second laser coagulation (by a non-contact 40 W point with a duration of the impulse of 0.5(-1). A permanent healing was achieved by the 14th day with the application of epithelializing ointment. PMID- 7485771 TI - [Abortions on demand in adolescents]. AB - The pregnancy is an integral part in the life of teenagers. When the conception was appeared the adolescents have to choose between interruption of delivery. Psychological status, contraceptive use and sexual behavior, motivation for abortion, operative methods and complications were observed in 463 teenagers, aged 13-17 years, who had an abortion voluntary in the last 6 years (1988-1993). We were found an abortion rate to 2.33% in our Dept. Obstet. Gynaecol. and a trend to increase the number of delivery in teenagers. The medical complications of abortions were: perforatio uteri complicata (0.65%) and endometritis acuta (9.29%), in cases with pregnancies over 16-20 g.a. This investigation was characterized from the teenagers as an stress and will be able to be complicate with a psychological disorders. PMID- 7485773 TI - [Endometriosis--the features of its clinical characteristics]. AB - The authors present the clinica; characterization to 100 women that operate in the Institute in Obstetrics and Gynaecology with histological diagnosis endometriosis, compare with 20 women, operate on the occasion of myoma uteri. The authors find that endometriosis happen more often than endometriosis externa (69%). The both most frequently are in combination with myoma uteri (68% and 52%). In the combination with Ca are a little (3%) with Ca corporis uteri. Sterility has in 14% on cases with endometriosis interna and twice more (32%) in the cases with endometriosis externa. Hypermenorrhea occurs in 43% on the cases and dysmenorrhea--in 25%. Asymptomatic endometriosis occurs in 68% on the cases. In conclusion the endometriosis is a unique disease with no characteristics symptomatic, that help us no much for the exact diagnosis. PMID- 7485774 TI - [Endometriosis in clinical practice]. AB - The authors are investigated the frequency of the cases of genital endometriosis, its most frequent localization and the choice of treatment on 864 women patients who have undergone operations connected with other gynaecological disorders and on 32 women patients, suffering sterility and proved endometriosis. Most frequent are the cases of combination between uterine myoma and endometriosis of the uterine wall-5.74% and with the patients who have undergone surgery for malignant disease between endometrium carcinoma and uterine endometriosis. The conservative treatment is the method of choice with the infertile women suffering a slight stage of endometriosis and the least conservative method is the surgical treatment, followed by a hormonal treatment resulting in the increase of the fertility of women who have the average stage of development of endometriosis. PMID- 7485775 TI - [Occupational damage to fertility in welders]. AB - The aim of the present study is to establish whether there is difference in the quality of the sperm of welders in comparison with members of other professions without potential occupational risks. There were examined 30 welders and a control group of 159 males. In all participants in the study by means of the case history and clinical examination the presence of other pathogenic factors besides the occupational influences was excluded. The percentage part of the males with deviations in the sperm mobility and morphology was significantly higher in the group of welders as compared with the control group (p < 0.001). There were discussed the possible mechanisms which can be responsible for the found changes. PMID- 7485776 TI - [The intrauterine ponderal index as a prognostic factor in fetal retardation]. AB - The authors aim at finding the connection between the intrauterine ponderal index (IUPol) and the neonatal ponderal index (NPol) and to estimate the efficiency of IUPol in the prediction of the asymmetric growth retardation in the new born babies with NPol below 2SD for the corresponding gestational age. 142 patients were examined. The foetus weight was calculated by the formula: EFBW = 1,4 x BPD x FL - 200, and the foetus length was calculated by the formula: 6,18 x 5,9 x FL. The authors concluded that the good sensitivity and specificity of the application of IUPol in the diagnostics of the asymmetric growth retardation need wider use of this method and the IUPol can be used for the elimination of the fetal retardation, because of the high predictable level of the negative test. PMID- 7485777 TI - [A comparative study of the potentials of a cytological method, the progesterone test and echography in the early detection of endometrial carcinoma in women in the postmenopause]. AB - An attempt was made to examine and compare the abilities of the cytological method, the progesterone test and the uterus sonography for early finding and screening of endometrial cancer in postmenopausal women. There were studied three groups of patients, each group of them consisted of 900 women, for the period of 10 years. The uterus sonography and progesterone test showed the best results. The cytological smears method was found not suitable for screening programme. PMID- 7485778 TI - [Neonatal bacterial sepsis. II. New developments in treatment and prevention]. PMID- 7485780 TI - [The cytological screening problems in cancer of the cervix uteri]. PMID- 7485779 TI - [LHRH analogs in gynecology]. PMID- 7485782 TI - [The ultrasonic diagnosis in utero of fetal arachnoid cysts]. PMID- 7485781 TI - [Non-trophoblastic tumors of the placenta]. PMID- 7485783 TI - [Deviatio ileopancreatica a modo Scopinaro and intrauterine fetal hypotrophy]. PMID- 7485784 TI - [Antiprogesterones--new prospects in induced abortion]. PMID- 7485786 TI - [The use of Synarel (nafarelin acetate--spray) in gynecology]. PMID- 7485785 TI - [The treatment of vulvovaginal candidomycosis with itraconazole (Sporanox)]. PMID- 7485787 TI - [The role of routine ultrasonic screening in the antenatal diagnosis of fetal retardation]. AB - The authors aim to evaluate the utility of ultrasound biometry in the diagnosis of IUGR. In a prospective study over 2.5 year period 142 pregnants at risk for IUGR were followed and their ultrasound results compared to those of 50 controls, investigated once after the 24 gestational week. The authors conclude that: BPD and FAC, evaluated only once, are unreliable in IUGR diagnosis; FAC is the most sensitive and specific index in onefold fetal biometry, particularly after the 34 gestational week; combing linear with non-linear parameters significantly increases. PMID- 7485788 TI - [Adolescent pregnancy and the development of pre-eclampsia--eclampsia]. AB - The authors aim to investigate the incidence and severity of pre-eclampsia- eclampsia in adolescent pregnancies (age under 17 years) and to evaluate the outcome of pregnancy and birth in this patients. With this purpose we compare obstetric and neonatal results in 528 adolescents aged between 13 and 17 years with the results of a control group of mothers aged between 20 and 24 years. We used clinical observation, laboratory findings, instrumental diagnostics and scoring systems. Data were computer-processed. Pre-eclampsia and eclampsia was diagnosed in 17 cases (3.22%), with mild forms prevailing (2.46%). Eclampsia developed in 4 pregnant (0.76%)-an incidence differing from that cited in the literature. We are impressed by the outmost severity of eclampsia in the adolescents, with 1 case of fatal outcome. Seven of the newborns were retarded. The most frequent indications to caesarean section were placental ablation and foetal distress syndrome. Operative delivery was performed in 41.18%. One case of foetal antenatal death and one death in the early neonatal period were observed, both in mothers with eclampsia. The authors conclude that the incidence of pre eclampsia and eclampsia in adolescents is lower than in general obstetric population, but if eclampsia develops, it is fulminant and in some cases is fatal for the mother or the foetus. This grave prognosis may be related to inappropriate antenatal care in this patient group. PMID- 7485789 TI - [The dynamics and trends in surgical delivery at the Maternity Home of the State Institutional Hospital]. AB - The study discusses the frequency of operative obstetric deliveries (cesarean section, forceps extraction and vacuum extraction) over a period of 18 years from 1976 till 1993. A total of 100,545 deliveries with 100,192 livebirths took place during this period with a decrease of the annual number of deliveries in the last years. The frequency of the various kinds of operative deliveries shows different tendencies. The frequency of cesarean section has increased from 4.87 per cent in 1976 to 19.58 per cent in 1993, this increase being particularly marked during the last 6 years. Meanwhile the vaginal operative deliveries have decreased their frequency. The decrease of forceps extraction is from 4.33 to 2.22 per cent (steadily about 2 per cent during the last years). The frequency of vacuum extraction has dropped from 1.89 per cent to 0 by the end of the 1980 and the beginning of the 1990. The tendencies of the development of obstetric operations are analysed as well as their influence on perinatal outcome. PMID- 7485790 TI - [The effects of intramuscular and oral prophylaxis with vitamin K on the coagulation status of low-weight newborn infants]. AB - The present prospective study carried out in 1993 in the SFH "Maichin Dom"- Sofia, includes newborn infants with birth weight below 2500 g. and aims to compare the effect of the prophylaxis of the haemorrhagic disease of the newborn by intramuscular and peroral application of vit. K. For this purpose some coagulation screening tests were followed out in intervals of 4-5 days during the first two weeks of life. These two groups are compared between one another and with non-prophylacted with vit. K healthy newborn babies. The results so obtained allow to make the conclusion that, on one side, the haemostasis of newborns with low birth weight is very labile and, on another, that the peroral and the intramuscular ways of prophylaxis with vit. K have the same effectiveness. PMID- 7485791 TI - [The direct immunofluorescence test for the diagnosis of Chlamydia trachomatis in gynecology]. AB - Using the direct immunofluorescent test Chlamyset antigen for detection of Chlamydia trachomatis antigens, the authors examined 176 gynecologic patients. The prevalence of chlamydial infection in this population was 26.1%. The method for detection of C. trachomatis using the direct immunofluorescent test is described in detail. The results of the study show the significant role of C. trachomatis as a cause of gynecologic pathology--it was detected in cases of endocervicitis in 28.8% in pelvic inflammatory disease in 34.7%, in sterility patients in 18.8%. The authors conclude that the proper diagnosis and management of these gynecologic disorders requires the routine use in the clinical practice reliable diagnostic tests for detection of C. trachomatis and the direct immunofluorescent test is very useful for this purpose. PMID- 7485793 TI - [The treatment of endometriosis in women with sterility. A prospective study over a 2-year period]. AB - Presented are the results of a 2-year prospective study in patients with no other causes than endometriosis proved by laparoscopy between Jan. 1991 and June 1992. Fifty five patients completed 6-9 months of therapy of Orgametril alone and 15 women were pregnant (27.27%). Forty one patients had electro surgery of endometriosis by the time of laparoscopy and 21 patients of them received Orgametril. After electro surgery alone 5 women were pregnant (25%) and of 21 women with complex therapy 10 were pregnant (47.6%). Control group of 79 patients without therapy had pregnancy rates for 6 patients (7.59%). No differences were observed between three groups of patients with treatment. Pregnancy rates were statistically lower in group without therapy. This study reported conception rates for 18 months after laparoscopy. PMID- 7485792 TI - [Our experience in treating pelvic inflammatory disease caused by Chlamydia trachomatis with the preparation roxithromycin]. AB - The authors are analyzing their results of the treatment with roxithromycin 150 mg of the pelvic inflammatory disease, caused by chlamydia trachomatis. 29 patients with proved chlamydial infection were treated. The most popular diagnosis was cervicitis. 23 patients were totally cured, 4 patients were with considerable better status after the treatment, and 2 of the patients were with insignificant amelioration of the status. A conclusion can be made, that the roxithromycin is a very good alternative of the existing treatment with doxycicline. PMID- 7485794 TI - [The incidence of clinical forms of female endocrine sterility in 1990-1993 at the III Gynecological Clinic of the Maternity Home of the State Institutional Hospital]. AB - The high frequency of the female endocrine sterility and the favorable therapy results are showing categorically the importance of the endocrine sterility as a medical and a social problem. For the period 1990-1993 in Third Gynaecological clinic--Sector of Gynaecological Endocrinology, were observed 584 patients with proved endocrine sterility. The present research has the goal to discuss the frequency of the different clinical forms of the endocrine sterility. The most frequent one is PCO--30.59%, followed by corpus luteum insufficiency--23.58%, and hyperprolactinaemia--20.05%. The results are interpreted on the basis of similar research for the period 1976-1985 with analog contingent. Conclusions are made for the essential directions of the diagnostic and therapy process in the clinic. PMID- 7485795 TI - [The medicosocial aspects of contraception for teenagers]. AB - The authors study and examine the influence of some medical and social factors on the teenagers use of contraceptives. The study is based on the questionnaires fulfill by 568 teenagers (aged 13-19). In the enquetes are shown the role of the family status of the parents the involvement of the family and the school in the sex education, and also are indicated the favorite teenager's contraceptive methods. The positive attitude on contraception and on this kind of researches is very characteristic for 94% of the analyzed teenagers. The authors made a conclusion on optimizing the use of modern contraceptives for teenagers. PMID- 7485796 TI - [Oligohydramnios]. PMID- 7485797 TI - [Legal abortion--different methods and complications]. PMID- 7485799 TI - [The historical stages in the conservative treatment of endometriosis]. PMID- 7485798 TI - [The place of vacuum extraction in modern obstetrics]. AB - A clinical trial was carried out on the use of vacuum extraction for delivery of 45 women. The aim of the study is to revive again this method for it was rarely used during the last years. The authors remind of the indications and necessary condition for the use of vacuum extraction and report about complications and the course of the placental period. Parallel to this, the 45 babies born with vacuum, are followed up according birth weight, gestational age, clinical status and laboratory data by birth, resuscitation after delivery and birth trauma. By summing up the results the authors divide women (and babies) in 3 groups: deliveries with vacuum extraction (VE), VE+ forceps, Vstimmulation and forceps, Conclusions are made that VE is a useful method nowadays if classical conditions and indications are followed. The babies born by VE are in good status without serious traumas. Vstimmulation + forceps is rejected as more harmful method. PMID- 7485800 TI - [A proposal for improving the classification of endometriosis]. PMID- 7485803 TI - [Primary carcinoma of the fallopian tubes--difficulties in its differential diagnosis and management]. PMID- 7485801 TI - [A subfascial abscess following cesarean section]. PMID- 7485802 TI - [Laparoscopic surgery in ectopic pregnancy]. AB - The authors present two cases of women with unruptured tubal pregnancies who were treated by methods of laparoscopic surgery. A salpingotomy as well as an aspiration of the pregnancy was performed. The operations reported are the first in gynaecological practice in Bulgaria and the operative technique is described. PMID- 7485804 TI - [The treatment of endometriosis genitalis externa with the preparation Zoladex from the firm of Zeneca, Great Britain, combined with an immune factor in sterility--a case report]. AB - Endometriosis is one of the main causes of female sterility, leading not only to subjective complaints, such as anovulatory menstrual cycles and LUF syndrome, but also to appearance of antisperm antibodies due to the increased number of macrophages, We present our experience in the treatment of endometriosis with the GnRH analogue Zoladex, product of Zeneca, as well as in the treatment of antisperm antibodies with methyiprednizolon acetate, Depomedrol of Upjohn. PMID- 7485805 TI - [The use of the preparation Betadiner in some forms of colpitis]. PMID- 7485806 TI - [Postmenopausal atrophic colpitis--its local treatment with low-dose estradiol]. PMID- 7485807 TI - [The effect of the diet on the iron status and incidence of anemia in pregnant women]. AB - The influence of dietary intakes during I and III trimester of pregnancy on iron status and anemia incidence [correction of prevalence] in 44 healthy pregnant women were studied. For evaluation of iron intake the probability approach calculations were used. Iron status was assessed by using multiple criteria--iron in serum, TIBC, transferrin saturation capacity and hemoglobin level. Iron deficient anemia is diagnosed in pregnant woman when hemoglobin level is less than 110 g/l and the least two of the other indices are abnormal. More than 70% of the women studied have low dietary intake of iron during pregnancy, but only 21% are anemic. The half of women with anemia are with iron deficient anemia. The results of iron intake may overestimate the risk of developing iron deficiency. Iron status during pregnancy is influenced by the dietary intake and diet structure, iron body stores and adaptive mechanisms. PMID- 7485808 TI - [Coagulation screening tests of low-weight newborn infants--deviations and dynamics]. AB - A prospective study was performed in 1993 in our hospital, including newborns with low birth weight and a control group of term infants. It aims to give information about their coagulation status and its dynamics. Abnormalities in the carried out screening tests were noticed in both groups during the first day of life. After the performed vit. K prophylaxis PT had almost the same dynamics in the two groups and almost reached the normal values in adults. aPTT remains highly prolonged in the group of low birth weight infants. Comparing the haemostasiological test levels between the group of low birth weight newborns, having received a prophylactic dose of vit. K i.m. and the corresponding group without vit. K prophylaxis, we established the faster normalizing of these levels in the first group. PMID- 7485809 TI - Effects of cerebral electrical stimulation on alcoholism: a pilot study. AB - Cerebral electrical stimulation (CES), born from research on electroanesthesia in the seventies, consists of the application of a pulsating current of small intensity (usually less than 1 mA, and below the threshold of perception) through the skull, e.g., in daily 30-min sessions. Claims of biological effectiveness (neurochemical, hormonal and EEG changes, naloxone-reversible analgesia in rats, etc.) and of clinical effectiveness (anxiety, depression, cognitive functions in alcoholics) have often relied on poorly controlled data. A recent controlled study in the treatment of opiate withdrawal has been positive. The present double blind controlled study compares active CES with sham stimulation in 64 alcohol dependent males. Over 4 weeks, both treatment groups improved significantly in most aspects. In the active treatment group additional significant improvement was observed in week-end alcohol consumption, and in two psychological measures: depression and stress symptoms index, but not in general drinking behavior. PMID- 7485810 TI - Alcohol schema acquisition in preschoolers: differences between children of alcoholics and children of nonalcoholics. AB - Cognitive schemas provide the structure within which children organize their knowledge and beliefs about the use of alcohol. The development of schemas about alcohol should be affected both by age and parental patterns of alcohol use. We examined differences in alcohol schema development among 139 male children of alcoholics (COAs) and 82 controls [children of nonalcoholics (NCOAs)] utilizing the Appropriate Beverage Task as an indicator of these processes. Overall, the vast majority of the sample identified at least one alcoholic beverage from photographs, even at age 3. COAs were more likely to identify at least one alcoholic beverage. With age controlled, COAs were better able to identify specific alcoholic beverages and correctly identified a larger number of alcoholic beverages. There was a trend for these children of alcoholic men to attribute more alcoholic beverage use to male adults than NCOAs. Moreover, differences in these children's attributions of alcoholic consummatory behavior were predicted by their parents' current consumption levels. Results provide evidence that alcohol schemas are detectable in early childhood and are more common in children from alcoholic homes. Discussion focuses on the potential relevance of these risk attributes to the development of more fully formed alcohol expectancies and to the later emergence of alcohol-related difficulty. PMID- 7485811 TI - Comparison of direct interview and family history diagnoses of alcohol dependence. AB - Using data from The Collaborative Study on the Genetics of Alcoholism, we compare direct interview diagnoses of alcohol dependence to those obtained by history from family members. Using a requirement of three or more positive implications by history, the specificity, sensitivity, and positive predictive values are 98%, 39%, and 45%, respectively. A logistic analysis found the gender of the relative and alcoholism in the informant to be significant, but not the gender of the informant. The partial odds ratio of a diagnosis at interview associated with a positive family history diagnosis was 13.6. The relationship between the informant and relative was significant, with negative reports from an offspring or mate more influential than a negative report from a parent or second-degree relative. We derived a recursive equation to combine a variable number of family history reports, wherein the probabilities associated with a single report are computed from the logistic analysis. This permits the use of family history information both as a proxy for an uninterviewed relative, as well as a second source of information to be used in the analysis of genetic family data. PMID- 7485812 TI - Hon v. Stroh Brewery Company: what do we mean by "moderate" and "heavy" drinking? AB - Although widely used, drinking terms such as "light,"' "moderate," and "heavy" are unstandardized and, as a result, public health messages using these terms may convey confusing information. As an initial attempt in providing such standardization, the present study surveyed public definitions for these terms. "Light" drinking was operationally defined as 1.4-2.4 drinks/day; "moderate" drinking was defined as 2.5-3.6 drinks/day; and "heavy" drinking was defined as 3.7 drinks/day and above. These ranges, however, were dependent on the respondent's gender, age, socioeconomic status, and especially the respondent's self-reported tolerance. Males had a higher threshold of consumption for "moderate" and "heavy" drinking than women. Older respondents likewise assigned a higher threshold for these terms than younger respondents. As respondent income increased, the threshold for "heavy" drinking decreased. The heaviest drinkers had a higher threshold for assigning the "heavy" label than did any other group. Ethnicity did not significantly affect these ranges, and religion only affected the threshold for "heavy" drinking. Because reseachers ultimately rely on terms such as "moderate" or ":heavy" to communicate their findings to colleagues and the public, it would seem apposite for them to agree on operational definitions for such terms. It would also seem appropriate to consider the operational definitions the public uses when referring to these terms to promote credibility of research findings and appropriate changes in behavior. PMID- 7485813 TI - Delayed P3A in abstinent elderly male chronic alcoholics. AB - Significant central nervous system toxicity in frontal brain regions has been demonstrated with chronic alcohol consumption both on autopsy and using neuropsychological testing. This study examined the latency of an objective and reproducible brain event-related potential measure of frontal cortex function in chronic elderly male alcoholics who were abstinent 3 months-2 years, a patient group in whom the central nervous system effects of chronic alcohol abuse are thought to be largest and most persistent. We examined the latency of the P3A event-related potential component, which reflects a frontal maximum orienting response to novel stimuli. Twelve elderly abstinent chronic alcoholic males and 11 elderly male controls were studied in an auditory and a visual paradigm, each of which included target, nontarget, and novel rare nontarget conditions. In both modalities, the P3A response to the novel rare nontarget stimuli was significantly delayed in the chronic alcoholics. P3B delays to the target stimuli were also present in the alcoholics, with the P3A and P3B effects being independent of each other. For both P3A and P3B, the effects were larger and more consistent in the visual compared with the auditory modality. Our conclusions are as follows: (1) both P3A and P3B latency delays are evident in elderly abstinent chronic alcoholics; (2) separate mechanisms are responsible for these effects; (3) these effects are more sensitively detected in the visual versus the auditory modality; and (4) delayed P3A latency may be an objective and reproducible index of the frontal cortex effects of chronic alcohol abuse. PMID- 7485814 TI - A test of Tiffany's cognitive model of drug urges and drug-use behavior. AB - Tiffany's (1990) cognitive model proposes that drug urges and drug use result from distinct (i.e., controlled versus automatic) cognitive processes. This study tested Tiffany's cognitive model utilizing innovative methods derived from the Multiple Resource Theory of cognitive psychology. Forty-two male and 42 female heavy drinking college students were assigned to 1 of 6 groups in two separate 1 (task) x 3 (treatment) factorial experiments in which half the subjects performed a math task while the other half performed a tracking task. Subjects received 1 of 3 treatments: Urge generation, "Drug" (Placebo) Consumption, or a Water control. The predictions were that urges would interfere with performance on the math task, and "drug" consumption would interfere with performance on the tracking task. The main dependent variables were measures of task performance. The results of this study do not clearly support the model; however, several suggestions for future tests of the cognitive model are discussed. Our findings highlight both the difficulty in testing the model, as well as opportunities for further integration of cognitive psychology and behavioral approaches to addictions. PMID- 7485815 TI - Alcohol consumption and physical fitness among young adults. AB - The relationship between alcohol consumption and physical fitness was analyzed using data from the baseline examination of the Coronary Artery Disease Risk Development In Young Adults study (CARDIA), a longitudinal study of the evolution of risk factors for cardiovascular disease in Black and White men and women aged 18-30 years. Two parameters of submaximal exercise treadmill testing were evaluated: time to heart rate 130 and duration of the treadmill test. In men and women qualified for treadmill testing, time to reach heart rate 130 was positively associated with ethanol intake. The relationship between fitness and ethanol intake persisted in sex-specific analysis after adjustment for age, ethnicity, systolic blood pressure, sum of subscapular and triceps skinfolds, and physical activity level. The magnitude and strength of the relationship were greater in women than in men. A negative interaction between ethanol intake and smoking status was evident. Linear regression coefficients in men were 0.29 sec to heart rate 130/ml ethanol/day in nonsmokers and -0.06 sec to heart rate 130/ml ethanol/day in smokers. Linear regression coefficients in women were 1.07 sec to heart rate 130/ml ethanol/day in nonsmokers and 0.23 sec to heart rate 130/ml ethanol/day in smokers. Men and women who smoked showed 31 sec longer time to heart rate 130, regardless of ethanol intake. Mean daily ethanol intake was positively associated with duration of treadmill testing in women, but weakly associated with duration in men. These results show increased physical fitness as ethanol intake increased in women and nonsmoking men and mildly decreased physical fitness as ethanol consumption increases among men who smoke.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7485816 TI - Variability of ethanol absorption and breath concentrations during a large-scale alcohol administration study. AB - Ethanol disposition was evaluated in 77 female and 97 male college seniors during an alcohol challenge study. All were regular drinkers who exceeded legal intoxication levels at least twice a month by history. A standard ethanol dose (females, 0.43 g/kg; males, 0.51 g/kg) was administered over 10 min, after a 4-hr fast, and breath alcohol concentrations (BrACs) were measured for 2 hr. Intersubject variability in BrACs was greatest early in the study, during ethanol absorption; the coefficient of variation decreased from 39% at 14 min to 14% at 125 min after the start of drinking. The time to peak BrAC varied from 10 to 91 min after the start of drinking (mean 39.6 min). Mean BrACs were significantly lower in females than males; mean peak BrACs were 0.054 g/210 liters in females and 0.058 g/210 liters in males (p = 0.031). The beta- and r-values for both genders were higher than those typically used in ethanol dose calculation formulas. Data are discussed to direct future research. The constants used in Widmark's formula need to be revised differentially for males and females in this population to reach specific target BrACs. Furthermore, substantial variability in absorption rates must be accounted for when assessing rising versus falling limb BrAC phenomena. PMID- 7485817 TI - Alcohol and other drug use among Hispanics in New York State. AB - Surveys have generally shown that Hispanics in the United States have lower than the national average rates of drinking, along with average rates of heavy drinking and drug use. Acculturation has been shown to be related to drinking and drug use. This study reports results from a large general population telephone survey conducted in New York State. Hispanic New York State residents were lower than the national average in drinking and heavy drinking when demographic variables were held constant. Gender differences in substance use were greater for Hispanics than for non-Hispanics, with males higher than females. Among Hispanic groups, Puerto Ricans were high and Dominicans were low in drug use. Among Hispanics, more frequent use of English was associated with higher drinking and drug use rates. Traditional Hispanic attitudes and closeness to country or origin were not related to substance use. PMID- 7485818 TI - Spatial learning deficits in preschool children of alcoholics. AB - When compared with nonalcoholics, chronic alcoholics and their children show significant deficits in the processing of visuospatial information. The literature supports two possible explanations of a visuospatial processing deficit in the child of an alcoholic (COA) when compared with the child of a nonalcoholic (NCOA). Either the COA may suffer cognitive disruptions produced by personal and social development within an alcoholic family, or the COA may inherit or very early develop alterations in central nervous system substrates of neurocognitive operations. The present study was designed to continue our examinations of visuospatial information processing differences and the source of these differences in COAs. An evaluation of very young subjects not only assisted in providing a more complete view of visuospatial processing across the COA's life span, but also helped explain why the deficit occurs. Thirty matched male and female preschool children, aged 35.8-51.6 months, served as participants. Fifteen children were COAs from families in which the biological father and two other relatives had an alcoholism history. The other group of 15 children were NCOAs. Each child performed a visuospatial learning task similar to the task used in previous studies of older COAs. The visuospatial learning performance of the preschool COAs was inferior to that displayed by preschool NCOAs. The patterns of correct, error, and nonresponses emitted by the preschool COAs and the interrelationships of these data closely resembled the data from our previous studies of older children, adolescent, and adult COAs.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7485819 TI - Reduced activities of thiamine-dependent enzymes in brains of alcoholics in the absence of Wernicke's encephalopathy. AB - The relative roles of alcohol per se, thiamine deficiency, and liver disease in the pathogenesis of alcohol-related brain damage have not been fully elucidated. In particular, the extent to which alterations of brain thiamine metabolism contribute to cognitive dysfunction in alcoholism in the absence of Wernicke's encephalopathy has not been established. In the present study, thiamine diphosphate-dependent enzymes were measured using standard spectrophotometric techniques in homogenates of brain tissue obtained at autopsy from eight alcoholic patients, all of whom died in hepatic coma without clinical or neuropathological evidence of Wernicke's encephalopathy and six nonalcoholic, age matched controls, matched for autopsy delay time and free, at the time of death, from gross malnutrition or other neurological or psychiatric disorders. Transketolase activities were reduced in cerebellum (by 35%, p < 0.01), thalamus (by 35%, p < 0.01), frontal cortex (by 22%, p < 0.01), temporal cortex (by 20%, p < 0.01), and prefrontal cortex (by 19%, p < 0.01). Activities of the pyruvate dehydrogenase complex were selectively reduced in prefrontal cortex by 25% (p < 0.01). Activities of alpha-ketoglutarate dehydrogenase were within normal limits in all brain regions of alcoholic patients. The generalized reductions of transketolase activity undoubtedly result from thiamine deficiency. Previous studies suggest that the presence of liver disease may exacerbate thiamine deficiency in alcoholics. A sustained loss of transketolase activity in brain could result in disruption of pentose shunt activity and concomitant reductions in reducing equivalents and lipid metabolism within the cell. The selective loss of pyruvate dehydrogenase activity in prefrontal cortex of alcoholic cirrhotics could relate to the phenomenon of hepatic coma.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7485820 TI - Brain proton magnetic resonance spectroscopy studies in recently abstinent alcoholics. AB - Chronic alcohol-dependent patients have reduced brain volumes and concomitant neurobehavioral deficits that may recover during abstinence. In 10 chronic alcoholic patients, using localized proton magnetic resonance spectroscopy, we found reliable increases during the first 3-4 weeks of abstinence in the concentrations within the superior cerebellar vermis of choline (Cho)-containing compounds relative to the neuronal marker, N-acetylaspartate (NAA). Lesser changes were observed following 1 month of abstinence, and in one of the patients studied longitudinally over 3 months, a marked reduction in the Cho/NAA ratio was associated with relapse. After detoxification, the Cho/NAA ratio correlated with a composite clinical impression of brain functions. The lowest Cho/NAA was observed in a patient with persisting alcoholic dementia, in striking contrast to reduced relative concentrations of NAA reported in dementia of the Alzheimer's type. Possible molecular explanations for these brain metabolic changes are discussed. PMID- 7485821 TI - Significant increase of blood alcohol by cimetidine after repetitive drinking of small alcohol doses. AB - To assess effects of repetitive alcohol drinking and pre-existing first-pass metabolism on the cimetidine-induced increase in blood alcohol concentrations, 20 healthy men (aged 20 to 40) of varied ethnicity and consuming less than 60 g alcohol per week underwent baseline quantitation of first-pass metabolism of alcohol. This was followed by oral administration of 0.6 g/kg ethanol given postprandially in 3 to 4 drinks spread over 135 min, before and after cimetidine (400 mg twice a day for 7 days). Blood alcohol concentrations were determined by breath analysis. First-pass metabolism was quantified by applying Michaelis Menten kinetics to blood alcohol curves after intravenous or oral administration of equal alcohol doses. At baseline, 15 subjects had a substantial first-pass metabolism (over one sixth of the dose); their alcohol levels increased with repeated doses with a mean peak of 27 +/- 3 mg/dl before and 39 +/- 5 after cimetidine (P < 0.01), an effect much greater and longer than after a single alcohol dose. Three subjects exceeded 50 mg/dl, the legal limit for driving in several countries. By contrast, in the five subjects with minimal first-pass metabolism, cimetidine did not increase alcohol levels. Thus, under conditions mimicking social drinking, cimetidine increased blood alcohol to concentrations known to impair psychomotor skills and they persisted at those levels over prolonged periods of time. In a minority of subjects, no such interaction was found, but their first-pass metabolism at baseline was absent or minimal and thus no inhibition by the drug was to be expected. PMID- 7485822 TI - Strategies for mapping and identifying quantitative trait loci specifying behavioral responses to alcohol. AB - Most responses to alcohol in both humans and animals are heritable, and this genetic sensitivity to ethanol is determined by multiple genes. However, the number of genes, their identities, and just how they determine susceptibility to the actions of alcohol are unknown. Herein, we describe a multistage strategy for mapping quantitative trait loci (QTLs) using recombinant inbred strains and F2 mice. Precise mapping of the chromosome positions of these QTLs should increase our understanding of the genetic causes for individual differences in behavioral sensitivity to alcohol by (1) identifying genomic markers associated with sensitivity to alcohol, (2) allowing the genes specifying behavior to be cloned by position, and (3) elucidating "candidate" genes demonstrating linkage to markers associated with behavioral responses to alcohol. Syntenic conservation between the mouse and human genomes should facilitate the eventual mapping and cloning of human homologs of these QTLs. Ultimately, cloning of these genes may allow the development of gene therapies or other therapeutic interventions for management or prevention of alcoholism and alcohol abuse. PMID- 7485823 TI - Classical and neoclassical approaches to the genetic analysis of alcohol-related phenotypes. AB - Theoretical descriptions are given for two breeding methods in animal genetics that might be of use in alcohol research. These methods are marker-based selection and marker-based development of congenic strains, both using DNA markers such as polymerase chain reaction-detectable polymorphisms as the criteria for breeding. Such designs would utilize these markers as indicators of adjacent Quantitative Trait Loci (QTL) that are influential on alcohol-related phenotypes. Issues in the logic and implementation of these methods, such as proximity of the markers and the QTL allele, are explored. A third method, development of congenic strains with phenotypic screening, is also described. This method is currently being used to create two sets of congenic lines on a C57BL/6 inbred mouse background. The criterion phenotype is locomotor activation to 1.5 g/kg (i.p.) ethanol. Data are reported on the success of transferring the activation phenotype from two strains, DBA/2Abg and MOLD/Rk-Abg, onto the nonactivated C57BL/6Abg background. The value of these methods in alcohol research is outlined with regard to both identification of relevant genes and for their use as tools in basic research on mechanism of alcohol action. PMID- 7485824 TI - Use of transgenics, null mutants, and antisense approaches to study ethanol's actions. AB - Behavioral and biochemical responses mediating ethanol's actions have been difficult to study in humans and animals because of their complex polygenic nature. Recent progress in the creation of new animal models using recombinant DNA technology has provided a set of genetic tools by which the role of specific candidate genes in ethanol's actions can be examined. These techniques include the creation of transgenic and null mutant mice, as well as manipulation of protein synthesis with antisense treatments. These techniques are reviewed, and their potential applications to alcohol research are discussed. PMID- 7485825 TI - A strategy for the identification of candidate genes for alcohol-related phenotypes and other human disorders using rapid polymerase chain reaction mapping of gene-based sequence-tagged sites. AB - We describe a method for the rapid identification and mapping of human genes, including those possibly contributing to disease and alcohol-related phenotypes. New human genes are identified from cDNA libraries through single-pass sequencing into the 3' untranslated (3'UT) regions of human brain cDNAs. Primers derived from the 3'UT region sequences [representing gene-based, sequence-tagged sites (STSs)] are used for polymerase chain reaction (PCR) analyses of the CEPH megabase insert yeast artificial chromosome (YAC) DNA pools. With this approach, approximately 18,000 megabase YACs can be screened and a single YAC identified using only 52 PCR reactions. The YAC localization in conjunction with other mapping techniques, such as PCR mapping to human chromosomes using somatic cell hybrids, allows identification of chromosomal band locations. In this manner, each gene can be associated with its own STS, which in turn specifies both a corresponding genomic clone and specific location in the genome. These locations can be compared with the purported locations of disease genes. The locations of the STSs can also be compared with those of Quantitative Trait Loci implicated for quantitative traits (e.g., alcohol-related phenotypes) on the basis of synteny between the mouse and human genes. Using this strategy, we found candidates for 78 human disease/syndrome genes among the first 220 genes mapped. PMID- 7485826 TI - Identifying alcoholism vulnerability alleles. AB - Identification of vulnerability alleles is one starting point for elucidating the web of interactions leading to alcoholism so that treatment and prevention can be improved. Heritability studies indicate that vulnerability alleles exist. Two molecular approaches for identifying them, direct analysis of candidate genes and genetic linkage, are highlighted in this review. Methodological problems that have been partially addressed and limitations for the applicability of the genetic findings are discussed. PMID- 7485827 TI - 1-week withdrawal from 8 weeks alcohol consumption protects the heart from sepsis induced dysfunction. AB - Gram-negative sepsis causes a depression of the myocardium such that ventricular function curves generated on isolated perfused hearts removed from septic rats are displaced downward and to the right of control. Alcohol consumption can also cause a depression of the myocardium, especially if the period of alcohol feeding is prolonged. However, even before overt changes in the myocardium can be measured as a result of alcohol consumption, chronic alcoholism can result in a potentiation of sepsis-induced cardiac depression (Am. J. Physiol. 250:H1857 H1863, 1991). The purpose of the present study was to determine if 1 week of withdrawal of alcohol from the diet after 8 weeks of alcohol consumption would reverse the potentiation by alcohol of sepsis-induced cardiac depression. Animals were fed an ethanol-containing diet in which ethanol contributed 36% of the total calories. Rats were fed this diet or a control liquid diet for 8 weeks, and then some animals were taken off the alcohol diet and placed on the control diet for 1 week. Sepsis was induced in control-fed, alcohol-fed or withdrawal animals by the administration of Escherichia coli into the dorsal subcutaneous space. Nonseptic animals received sterile saline in this space. The following day animals were anesthetized, and the hearts were removed and studied as isolated working hearts. Hearts removed from septic and alcohol septic animals showed severe depression of cardiac contractile performance. Hearts from the withdrawal group, however, were less compromised by sepsis and showed only a few signs of cardiac dysfunction. Withdrawal from alcohol for 1 week thus resulted in protection of the heart from sepsis-induced cardiac depression. PMID- 7485828 TI - Continuous exposure of cultured rat cerebellar macroneurons to ethanol-depressed NMDA and KCl-stimulated elevations of intracellular calcium. AB - This series of experiments measured ethanol-induced changes in levels of free intracellular calcium. Cerebellar macroneurons, harvested from rat embryos on embryonic day 17, were cultured in the presence of 75 mM ethanol for 24, 48, or 96 hr. Intracellular calcium concentrations in control and ethanol-exposed neurons did not differ after 24 hr, but they were significantly elevated in the neurons exposed to ethanol for 48 or 96 hr. Similarly, increases in intracellular calcium elicited by stimulation with 50 microM NMDA were not significantly different in control and ethanol-exposed neurons after 24 hr. After 48 and 96 hr, however, NMDA-stimulated increases in intracellular calcium levels in control neurons were significantly greater than in the ethanol-exposed neurons. These results showed that, when calcium levels were elevated by prolonged exposure to ethanol, the neurons were significantly less responsive to NMDA stimulation. Increases in intracellular calcium elicited by stimulation with 30 mM KCI were not significantly different in the control and treated neurons after 24 and 48 hr of ethanol exposure. After 96 hr of exposure to ethanol, however, there was a significant increase in intracellular calcium levels in control neurons following KCI stimulation, but not in the ethanol-exposed neurons. The fact that neuronal responses to KCI stimulation were depressed only following 96 hr of exposure to ethanol makes it unlikely that voltage-regulated channels were the primary mediators of the ethanol-induced elevations in intracellular calcium in chronically exposed neurons. PMID- 7485830 TI - Apoptosis and bcl-2 protein expression in experimental alcoholic liver disease in the rat. AB - We used the intragastric feeding rat model to investigate the relationship between severity of alcoholic liver injury, apoptosis, bcl-2 protein expression, and lipid peroxidation. Rats were fed ethanol with different dietary fats (saturated fat, corn oil, and fish oil) for a 1-month period. Apoptosis was evaluated using an immunohistochemical method, and flow cytometry. Bcl-2 protein concentrations in liver were evaluated by Western blot analysis and lipid peroxidation by measurement of conjugated dienes. Pathological changes (fatty liver, necrosis, and inflammation) were present in corn oil-ethanol and fish oil ethanol groups only. The highest number of apoptotic cells were seen in the group of rats exhibiting liver injury. The fish oil-ethanol-fed group had the highest concentrations of bcl-2 protein; this protein was localized in the bile duct epithelial and inflammatory cells. A significant correlation was seen between bcl 2 protein assessed densitometrically and the number of inflammatory cells/mm2 (r = 0.78, p < 0.02) and conjugated diene levels (r = 0.82, p < 0.01). Increased numbers of apoptotic cells were seen in rats developing ethanol-induced pathological liver injury. Increased bcl-2 protein concentration are associated with the presence of inflammatory cells and lipid peroxidation. PMID- 7485829 TI - NMDA prevents alcohol-induced neuronal cell death of cerebellar granule cells in culture. AB - Neuronal cell loss is one of the most debilitating effects of alcohol exposure during development of the nervous system. In this study, primary cultures of neuronal cells (cerebellar granule cells) were used to examine mechanisms of alcohol-induced neuronal cell death. Previously, we established that (Pantazis et al., Alcohol Clin Exp Res 17:1014-1021, 1993): (1) alcohol exposure caused neuronal cell death in cultures of cerebellar granule cells and this cell loss was both time-dependent and dose-dependent; and (2) the vulnerability of cerebellar granule cells to alcohol-induced loss changed with the length of time the cells were in culture before initiating alcohol exposure-that is, younger cultures (1 day in vitro) were much more susceptible to alcohol-induced neuronal cell death than older cultures (4 or 7 days in vitro). The primary goal of the present study was to examine the potential role of the NMDA receptor in alcohol induced death of cerebellar granule cells in culture. Experiments were performed to test the hypothesis that the alcohol-induced death of cerebellar granule cells can be prevented or reduced by NMDA treatment. Our results indicate that stimulation of the NMDA receptor has a neuroprotective effect and can significantly reduce the alcohol-induced neuronal cell death of newly established cerebellar granule cell cultures. This neuroprotective effect of NMDA is blocked by 2-amino-5-phosphonovalerate, a competitive inhibitor of the NMDA receptor, confirming that this neuroprotective effect is mediated via the NMDA receptor. This is the first report that alcohol's neurotoxic effect can be ameliorated by activation of the NMDA receptor. PMID- 7485831 TI - Rabbit liver class III alcohol dehydrogenase: a cathodic isoform with formaldehyde dehydrogenase activity. AB - Electrophoresis of rabbit liver homogenate on starch gel followed by activity staining revealed multiple forms of alcohol dehydrogenase (ADH) which, based on their electrophoretic mobilities, had been tentatively labeled as class "I," class "II," and class "III" ADHs. The class II enzyme has now been purified to homogeneity by ion exchange and affinity chromatography and, except for an isoelectric point of 7.7, closely resembles human class III ADH. It is a homodimer of molecular weight near 80,000 with a similar amino acid composition and comparable kinetic parameters for the oxidation of primary alcohols. Like the rat, human, and Escherichia coli class III ADHs, the rabbit enzyme is a glutathione-dependent formaldehyde dehydrogenase, and catalyzes the oxidation of S-hydroxymethylglutathione and the hemithiolacetal of 8-thiooctanoic acid. Ethanol up to 3 M does not saturate the enzyme, whereas longer chain primary alcohols exhibit Michaelis-Menten kinetics. PMID- 7485832 TI - Effect of ethanol on plasma and hepatic insulin-like growth factor regulation in pregnant rats. AB - Alcohol consumption during pregnancy has been shown to have profound developmental and behavioral effects on the fetus; however, the specific cause of these abnormalities remains unknown. These studies examined the consequences of chronic ethanol exposure during pregnancy on the regulation of maternal plasma and hepatic insulin-like growth factors (IGFs), and their associated plasma binding proteins (IGF-BPs). Ad libitum, pair, and ethanol-fed rats were fed a commercial liquid diet containing either ethanol or isocaloric maltose-dextrin from day 2 of pregnancy through parturition and killed 6 hr postpartum. Maternal plasma IGF-1 concentrations were reduced 51% in ethanol, compared with pair-fed mothers, with a corresponding 20% reduction in hepatic IGF-1 mRNA levels. In contrast, plasma IGF-2 concentrations were increased approximately 100% in ethanol-fed mothers. Whereas the smaller forms of the IGF-binding protein subunits (24 kDa and 32-29 kDa) were not affected by ethanol treatment, a significant reduction was observed in the binding subunit of IGF-BP3 (45-40 kDa) in ethanol-exposed mothers. These results suggest that alterations in plasma and hepatic IGF regulation may contribute to changes in maternal and placental metabolism and hormone regulation during pregnancy, which may in turn contribute to the intrauterine and postnatal growth retardation observed in prenatally ethanol-exposed offspring. PMID- 7485833 TI - Behavioral effects and pharmacokinetics of propofol in rats selected for differential ethanol sensitivity. AB - High- and low-alcohol sensitivity (HAS and LAS) rats have been selected for their differences in ethanol-induced sleep time. The rats also differ in sensitivity to pentobarbital, halothane, isoflurane, and enflurane. To determine if this sensitivity extended to propofol, the anesthetic requirements were measured. In this study, the sleep time and the tissue levels of propofol at awakening, as well as the pharmacokinetics, were evaluated. Propofol was administered intravenously. For one group of rats, sleep times were measured; blood and brain samples were taken at awakening. Blood samples were collected in another group of rats at frequent intervals from 0 to 90 min after injection. Propofol concentration of the samples was determined by gas chromatography. The pharmacokinetic analysis was performed using a nonlinear least-squares regression program. Sleep time was not different; however, blood and brain propofol levels at awakening showed a small, but significant difference between HAS and LAS rats. Propofol blood concentration-time curve data were fitted to a three-compartment model. Pharmacokinetic parameters were also not different between the rat lines. However, sleep time was 50% longer in female rats than male rats in both strains (p < 0.0001). The rates of propofol clearance were slower in female rats, because of different rates of disappearance from the second compartment. The observations suggest that the genetic selection for ethanol sensitivity selection for propofol sensitivity was not nearly as intense and presumably involves some different genes. These two central nervous system depressants would seem to differ significantly in their mechanism of action.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7485834 TI - Structural reorganization in the supraoptic nucleus of withdrawn rats following long-term alcohol consumption. AB - We have recently shown in the supraoptic nucleus (SON) of the rat that prolonged ethanol consumption induces cell degeneration and enlargement of the surviving neurons and of their subcellular organelles. We analyzed the SON of withdrawn rats to evaluate whether it displays any evidence of morphological reorganization following abstinence from ethanol, inasmuch as in this condition the ethanol induced changes in the plasma levels of neurohormones and plasma osmolality are no longer detectable. A group of 18-month-old withdrawn rats was compared with age-matched, pair-fed control and ethanol-treated rats. To differentiate between the effects of withdrawal and the effects of rehydration, a group of 18-month-old rehydrated rats was also included in this study and compared with age-matched, pair-fed control and dehydrated rats. We estimated the volume of SON, and the total number and mean volume of its neurons. The cross-sectional areas of the vasopressinergic and oxytocinergic populations were also evaluated. At the ultrastructural level, we determined the volumes and surface areas of the rough endoplasmic reticulum and Golgi apparatus, and the volumes of neurosecretory granules and nucleoli. In withdrawn animals, the total number of SON neurons was smaller than in controls, although the neuronal volume was greater. The number of SON neurons did not differ between withdrawn and ethanol-treated rats, despite the reduced volume of SON in the former animals. The decrease of SON volume correlated with and was caused by a reduction in the volume of SON neurons and in the size of the organelles involved in neuro-hormone synthesis.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7485835 TI - Chronic ethanol consumption alters effects of ethanol in vitro on brain membrane structure of high alcohol sensitivity and low alcohol sensitivity rats. AB - In this study, we examined if differences in initial membrane sensitivity to ethanol were associated with development of membrane tolerance to ethanol. High Alcohol Sensitivity (HAS) and Low Alcohol Sensitivity (LAS) rats were administered a 15% ethanol solution in water as the sole source of fluid for 30 days. The amount of ethanol consumed per day did not significantly differ between the HAS and LAS rats. Development of membrane tolerance to in vitro effects of ethanol has been previously reported for bulk membrane fluidity and protein-lipid interaction. Our data expands the understanding of "membrane tolerance" phenomenon to protein distribution and bilayer interdigitation. We also introduce genotype-dependent and genotype-independent properties of the membrane tolerance to ethanol. ethanol treatment produced genotype-dependent and genotype independent membrane tolerance to ethanol. The in vitro effects of ethanol on synaptic plasma membrane (SPM) protein distribution and lipid bilayer interdigitation were abolished or decreased in the SPM of chronic ethanol-treated HAS rats, as compared with the SPM of HAS control rats (genotype-dependent tolerance). Protein distribution and bilayer interdigitation were not affected by ethanol in vitro in either chronic ethanol-treated or control LAS rats. Genotype independent tolerance to ethanol in vitro was observed for SPM annular and bulk bilayer fluidity in chronic ethanol-treated HAS and LAS rats. It is concluded that initial sensitivity to ethanol contributes to the development of membrane tolerance to ethanol in HAS and LAS rats. PMID- 7485836 TI - Possible role of striatal adenosine in the modulation of acute ethanol-induced motor incoordination in rats. AB - Several reports from our laboratory have suggested the involvement of the brain adenosinergic system in ethanol-induced motor incoordination (EIMI). This study is an extension of the previous work and pertains to the evaluation of the role of the striatal adenosine in EIMI in male Sprague-Dawley rats. Using the motor incoordination induced by 1.5 g/kg of ethanol (ip) as a test response, the possible behavioral interactions between ethanol and adenosine agonists and antagonists in the striatum were investigated. Intrastriatal (IST) administration of adenosine A1-, A1 = A2-, and As-selective agonists, R(-)N6-(2 phenylisopropyl)adenosine (R-PIA), 5'-N-ethylcarboxamido-adenosine (NECA), and 5' (N-cyclopropyl)-carboxamidoadenosine, respectively, significantly and dose dependently accentuated EIMI when evaluated by rotorod test, suggesting the striatal adenosinergic modulation of EIMI. No significant change in normal motor coordination was noted, even when the highest IST doses of adenosine agonists were followed by saline instead of ethanol, suggesting that the observed behavioral interactions of these drugs were selective to ethanol. Hippocampus, which is known not to be involved in the normal motor functions, was selected as a control brain area because of the presence of high density of adenosine receptors, as well as the high levels of adenosine. Intrahippocampal NECA failed to alter EIMI, indicating the specific role of striatal and not hippocampal adenosinergic system in the modulation of EIMI. The potentiating effects of adenosine agonists N6-cyclohexyladenosine (CHA) and CGS-21680 on EIMI were blocked by adenosine A1- and A2-selective antagonists, 8-cyclopentyl-1,3 dipropylxanthine and 3,7-dimethyl-1-propargylxanthine, respectively, suggesting the participation of specific adenosine receptors in this functional interaction. A role for the adenosine A1 receptor in the striatal adenosinergic modulation of EIMI was favored based on the rank-order potency of adenosine agonists. IST pretreatment with pertussis toxin (PT), but not with PT beta-oligomer, nearly completely eliminated the accentuation of EIMI by CHA, further supporting the favored role of adenosine A1 receptors in EIMI. Histological and IST [3H]R-PIA distribution data confirmed that the observed behavioral effects were caused by exclusive striatal distribution of intrastriatally microinjected drugs. Data obtained suggested modulation of acute EIMI by striatal adenosine receptor mediated mechanism(s) and the coupling of these adenosine receptor to the PT sensitive Gi protein. PMID- 7485837 TI - Acute ethanol impairs spatial memory but not stimulus/response memory in the rat. AB - The present studies investigate how acutely administered ethanol (ETOH) affects the use of spatial memory. Previous electrophysiological results have shown that acutely administered ETOH alters the firing of hippocampal neurons and that prenatal or chronic exposure to ETOH produces deficits on spatial learning tasks, tasks dependent on the hippocampus. In contrast, it has also been demonstrated that acutely administered ETOH does not impair spatial localization. In the current studies, rats were trained to perform a spatial or stimulus/response task under saline. After training, rats were injected with ETOH, and the use of spatial or stimulus/response memory was tested. Acutely administered ETOH impaired the use of spatial memory, but not the use of stimulus/response memory. Because the use of spatial memory requires an intact hippocampus, the present studies suggest that acutely administered ETOH selectively impairs behaviors dependent on the hippocampus. PMID- 7485838 TI - Regional differences in the effects of chronic ethanol administration on [3H]zolpidem binding in rat brain. AB - A strong association has been observed between [3H]zolpidem binding and the presence of gamma-aminobutyric acid (GABAA) receptor mRNA for alpha 1-, beta 2-, and gamma 2-subunits in specific brain regions. This correlates with observed sensitivity of individual neurons to zolpidem and ethanol in these same regions. Previous studies using homogenate binding approaches showed small alterations in [3H]zolpidem binding levels after chronic ethanol exposure. This study was undertaken to ascertain if there is regional specificity of the effects of chronic ethanol administration on [3H]zolpidem binding levels. Chronic ethanol administration induced small, but significant alterations in [3H]zolpidem (5 nM) binding in the inferior colliculus, substantia nigra, and the medial septum. [3H]Zolpidem binding was increased in the inferior colliculus and substantia nigra, and decreased in the medial septum. No significant differences in [3H]zolpidem binding were noted in any other brain area analyzed, including the cortex and cerebellum. These findings show that chronic ethanol administration has small effects on [3H]zolpidem binding, although they occur in a site-specific and bidirectional manner. Moreover, there is no correlation between changes in [3H]zolpidem binding and alterations in GABAA receptor subunit expression. PMID- 7485839 TI - Ethanol as a possible cofactor in the development of murine AIDS. AB - Chronic ethanol (EtOH) abuse in humans leads to a variety of immunomodulatory events that can alter resistance to a number of infectious agents. Whether alcohol abuse affects the susceptibility to human immunodeficiency virus infection or the subsequent development of acquired immune deficiency syndrome (AIDS) is a matter of extreme importance; however, available information in humans or animal models is limited. The goal of this study was to evaluate the effect of chronic EtOH feeding in mice on the development of immunodeficiency in the murine model of AIDS (MAIDS). C57BI/6 mice were placed on the Lieber-DeCarli liquid EtOH diet (25% or 31% total caloric intake) or a nutrient-matched isocaloric liquid control diet. Seven days later, mice were infected with the LP BM5 murine leukemia virus mixture, and groups of infected and noninfected mice were assayed at defined time points postinfection for antigen-specific and nonspecific immune responses. In the absence of retroviral infection, chronic EtOH feeding (5-8 weeks) led to reductions in spleen weights, compared with isocaloric controls. In spite of reduced spleen size, mitogenic responses of spleen cells to concanavalin A (ConA) and lipopolysaccharide (LPS) were elevated in EtOH-fed mice, as compared with mice fed the control diet. Chronic EtOH feeding also enhanced the allogeneic mixed lymphocyte response and increased antigen-specific priming of both B-cells and CD4+ T-cells to the antigen, sheep red blood cells. In MAIDS-infected mice, chronic EtOH feeding delayed but did not prevent the onset of virus-induced immunodeficiency and MAIDS-induced autoantibody synthesis.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7485840 TI - Changes in blood alcohol levels as a function of alcohol concentration and repeated alcohol exposure in adult female rats: potential risk factors for alcohol-induced fetal brain injury. AB - Fetal alcohol syndrome and alcohol-related birth defects are the result of heavy maternal alcohol consumption during gestation. The magnitude of deficit manifested by the offspring is invariably a consequence of several risk factors that may result in high peak blood alcohol concentrations (BACs), such as the duration, timing, or pattern of alcohol consumption. In addition, the alcohol content of the consumed beverage may play a role in determining offspring developmental consequences. Because higher BACs are positively correlated with risk and severity of brain injury early in postnatal life, initially it was important to determine how BAC is influenced by alcohol concentration and whether that influence is constant over repeated alcohol treatments. Groups of female Sprague-Dawley rats received daily intragastric intubations of 5 g/kg alcohol in one of several concentrations: 45% (v/v), 30% (v/v), 22.5% (v/v), or 15% (v/v) for a duration of 18 consecutive days. Blood samples were taken at various times postintubation on days 3, 8, 13, and 18 of treatment, and analyzed by headspace gas chromatography. Multivariate analyses of peak BAC, average BAC, and time to reach peak BAC revealed some noteworthy results. First, peak BAC and average BAC were significantly lower in the 45% group, compared with the other concentration groups, whereas this group also took a longer time to reach peak BAC than the other three groups. Second, peak BAC and average BAC were higher on the last day of treatment than any of the other treatment days. These results suggest that alcohol concentration and repeated alcohol exposure can influence BAC and, as such, are important risk factors to be considered in the appraisal of alcohol induced fetal brain injuries. PMID- 7485842 TI - Validated fluorimetric HPLC analysis of acetaldehyde in hemoglobin fractions separated by cation exchange chromatography: three new peaks associated with acetaldehyde. AB - Stable hemoglobin-acetaldehyde adducts present in hemoglobin fractions separated by polyaspartic acid cation exchange chromatography were quantified by fluorimetric HPLC. The fluorescent species eluted from the HPLC was confirmed by mass spectrometry to be consistent with the expected product from reaction of acetaldehyde, 1,3-cyclohexanedione (CHD), and ammonium ion. Hemolysate (2.2 mM hemoglobin) was incubated in equivalent volumes of either phosphate-buffered saline or 5 mM acetaldehyde at 37 degrees C for 30 min and washed three times with H2O to remove free acetaldehyde and labile adducts before the injection of 14.7 mg hemoglobin onto the cation exchange column. Fluorimetric HPLC analysis of hemolysate samples either with or without in vitro reaction with acetaldehyde revealed that most acetaldehyde resides in the hemoglobin A0 fraction. The reaction with acetaldehyde in vitro resulted in a significant increase in fast eluting minor hemoglobin species on cation exchange chromatography concomitant with increased acetaldehyde in the HbA1a+b, HbA1c, and HbA1-AcH fractions. We report three new cation exchange chromatographic peaks after reaction with acetaldehyde: HbA1-AcH-3, HbA1c-1, and HbA0-1. Each new peak was found to associate with a significant quantity of CHD-reactive acetaldehyde. These experiments provide additional evidence that stable adducts form between acetaldehyde and hemoglobin and that these adducts occur in multiple hemoglobin species separated by cation exchange chromatography. Further characterization and structural assignment of these species are warranted in view of their potential utility as markers for ethanol intake. PMID- 7485841 TI - Cellular signals in alcohol-induced liver injury: a review. PMID- 7485844 TI - Alcohol-metabolizing enzyme polymorphisms and alcoholism in Japan. AB - The liver enzymes, alcohol dehydrogenase (ADH) and aldehyde dehydrogenase (ALDH), which are responsible for the oxidative metabolism of ethanol, are polymorphic in humans. Cytochrome P450IIE1, an ethanol-inducible isozyme of liver microsomal P450, is also important in ethanol metabolism. Genetic polymorphisms in the 5' flanking region of the human cytochrome P450IIE1 gene have recently been reported. We hypothesized that the polymorphisms of ADH, ALDH, and P450IIE1 modify the susceptibility to development of alcoholism. We determined the genotypes of the ADH2, ALDH2, and P450IIE1 loci of 96 Japanese alcoholics and 60 healthy male subjects, using leukocyte DNA by the restriction fragment-length polymorphism by polymerase chain reaction. The alcoholics had significantly higher frequencies of the ADH2(1) and ALDH2(1) alleles than did the healthy subjects. No significant difference in the frequency of the P450IIE1 genotype was observed between the alcoholics and the healthy subjects. In conclusion, genetic polymorphisms of the ADH and ALDH genes, but not of the P450IIE1 gene, influence the risk of developing alcoholism in Japanese. PMID- 7485843 TI - Conditioned taste aversions induced by alcohol and lithium in rats selectively bred for ethanol neurosensitivity. AB - Rats that were selectively bred for differences in alcohol-induced sleep time (alcohol neurosensitivity) were tested for differences in formation and extinction of alcohol- and LiCl-induced conditioned taste aversions. Male rats bred for high, control, or low alcohol sensitivity (HAS, CAS, and LAS rats, respectively) were deprived of water and given daily 30 min access to water for a baseline period of 7 days. Rats were then given a novel 0.125% sodium saccharin solution, followed by an intraperitoneal injection of either saline, 2 g/kg of ethanol (at 10% w/v), or 50.9 mg/kg of LiCl (0.15 M) on 3 conditioning days. Each saccharin exposure was followed by a recovery day of access to water. The ethanol induced saccharin aversion extinguished more rapidly in LAS rats than in CAS or HAS rats (p < 0.05), but LiCl conditioned equivalent aversions in each group. Also, ethanol injection results in large differences in observed resting behavior in these rats (HAS > CAS > LAS), but LiCl injection produced no reliable group differences in resting. The weaker alcohol-induced taste aversion in LAS rats accords with their previously measured higher oral consumption of alcohol (Kulkosky et al., Alcoholism 17:545-551, 1993) and the idea that alcohol intake is limited by an expectancy of postingestive consequences. The weaker ethanol induced aversion in LAS rats reflects selective breeding of an alcohol-specific trait and not a general difference in aversive conditioning or chemical neurosensitivity. PMID- 7485845 TI - Task persistence after alcohol consumption among children of alcoholics. AB - Two studies examined task persistence after alcohol consumption among young adult offspring of alcoholics [children of alcoholics (COAs)] and young adult offspring of nonalcoholics (NonCOAs). In study I, heavy drinking college seniors (n = 294) participated in a laboratory alcohol challenge in the context of a baseline assessment for a longitudinal study. Subjects completed a computerized administration of the Matching Familiar Figures Test in two halves: the first half sober, and the second half approximately 50 min after consuming a 0.8 ml/kg body weight dose of ethanol. COAs demonstrated significantly greater acceleration of response latencies after consumption of alcohol compared with NonCOAs. In study II, moderate-to-heavy drinking volunteers (n = 149) were recruited for a one-time laboratory alcohol challenge, and completed a circle-tracing task three times approximately 50 min after consuming a 0.8 ml/kg body weight dose of ethanol. COAs displayed significantly less persistence on the second and third circle-tracing trials compared with NonCOAs. Data are discussed in relation to models of impulsivity and differential alcohol sensitivity among COAs. PMID- 7485846 TI - Detection of relapses in alcohol-dependent patients using carbohydrate-deficient transferrin: improvement with individualized reference levels during long-term monitoring. AB - The present study examined whether the sensitivity of carbohydrate-deficient transferrin (CDT) in serum, a biochemical marker of recent excessive alcohol consumption, could be improved during long-term monitoring by introducing individualized cut-offs between normal and elevated CDT levels. Alcohol-dependent male outpatients (n = 22), trying to abstain from alcohol for 6 months, were monitored by comparing weekly measurements of CDT with self-reports of alcohol consumption three times/week and daily urinary levels of 5-hydroxytryptophol (5 HTOL), a new marker of recent alcohol intake. The method used to calculate cut offs was based on the intraindividual variation in CDT not dependent on excessive alcohol consumption or analytical variations. An increase in CDT exceeding the minimum level for each patient by 3 and 4 times the mean coefficient of variation for healthy social drinkers (i.e., by 30% and 40%) was compared as an indication of alcohol consumption, even if the value did not exceed the conventional cut off. By using individualized CDT cut-off points, 68 and 41 episodes of drinking were detected in the patients with the cut-offs of > 30% and > 40%, respectively, as compared with 25 with the conventional limit. Most episodes could be verified clinically and/or by elevated urinary 5-HTOL levels during the 2-week period preceding each serum sampling. The results suggest that the possibility to detect relapses by CDT can be improved during long-term monitoring of alcohol-dependent outpatients by introducing individualized cut-off points between normal and elevated CDT levels. PMID- 7485847 TI - Group similarities in beliefs concerning the causes of alcohol problems and the effectiveness of means of overcoming problem alcohol use. AB - Four hundred and eight respondents, who did or did not know persons close to themselves with alcohol problems, from four different racial/ethnic groups (with one group, Filipinos, categorized in terms of whether they were born in the Philippines or in the U.S.) rated the importance of 19 different causes of problem alcohol use and 22 different means of overcoming problem alcohol use. Rank-order correlations across sexes, persons differing in exposure to problem drinkers, and ethnic groups were nearly all in the 0.90's regarding causation. Correlations of Caucasians' ratings with those of other groups were high, but only in the 0.60's and 0.70's, chiefly because Caucasians were more prone to regard problem alcohol use as hereditary and as a disease, and less likely to believe problem use to result from social factors. All across-group correlations concerning beliefs about the effectiveness of means of overcoming problems were in the 0.90's. Some of the means of treatment regarded most highly by all groups have not been demonstrated to be effective. PMID- 7485848 TI - Beta-carbolines in chronic alcoholics undergoing elective tumor resection. AB - The prevalence of chronic alcoholism in patients with carcinomas of the upper digestive tract exceeds 60%. The patient's history and laboratory markers, preoperatively, are often not sensitive or specific enough to detect alcohol dependent patients, preoperatively, who are at risk of developing alcohol withdrawal syndrome (AWS) during their postoperative intensive care unit (ICU) stay. Previously, it was found that plasma norharman was elevated in chronic alcoholics, suggesting marker characteristics for chronic ethanol misuse and possibly alcohol dependence. We investigated whether beta-carbolines (i.e., harman and norharman) were different between chronic alcoholics and nonalcoholics with carcinoma, and how the levels change in alcohol-dependent patients during their hospital stay. Ninety-seven patients with oral, pharyngeal, laryngeal, or esophageal carcinomas were evaluated regarding their drinking habits. Sixty patients were transferred to the ICU following tumor resection. Chronic alcoholics met the DSM-III-R and ICD-10 criteria for alcohol dependence or chronic alcohol abuse/harmful use. The daily ethanol intake in chronic alcoholics was > or = 60 g. Blood samples were collected on admission to the hospital, preoperatively, on admission to the ICU and on days 2, 4, and 7 in the ICU. Harman and norharman were determined by HPLC. Elevated norharman was found in chronic alcoholics on admission to the hospital, whereas harman did not differ between groups. On admission, the area under the receiver operating characteristics curve was significantly larger for carbohydrate-deficient transferrin and preoperatively for norharman. The preoperative norharman levels were significantly correlated with the period of mechanical ventilation and the length of ICU stay. Postoperatively, norharman decreased in all patients, except a group of 11 alcohol-dependent patients who developed AWS during their ICU stay. The finding that elevated norharman levels were found in chronic alcoholics on admission to the hospital and preoperatively supports the view of a specific marker for alcoholism. Preoperative norharman was superior to carbohydrate deficient transferrin and was associated with a prolonged ICU stay and a prolonged period of mechanical ventilation. Further studies are required to determine whether norharman aids in the preoperative diagnosis of chronic alcohol misuse with respect to the prevention of postoperative complications. PMID- 7485849 TI - Effects of hostility on alcohol stress-response-dampening. AB - The purpose of this study was to compare the stress-response-dampening (SRD) effect of alcohol in hostile and nonhostile men based on a combined score of four subscales of the Cook-Medley Hostility Scale. Subjects were 72 male social drinkers. Subjects' cardiac interbeat-interval, and systolic and diastolic blood pressure reactivity to a situational stressor were measured following the consumption of either alcohol, no alcohol, or an active placebo beverage. Results demonstrated that hostile men evinced lower heart rate and systolic blood pressure (SBP) reactivity to the stressor when given alcohol, compared with intoxicated nonhostile subjects, and lower reactivity relative to all other groups, with the exception of SBP in the nonhostile controls. These results allow for speculation that hostile men may be more likely than controls to experience possible SRD effects of alcohol and thus, perhaps, be predisposed to increased alcohol consumption when under stress. PMID- 7485850 TI - Predictors of mortality in alcoholic men: a 20-year follow-up study. AB - This study investigates the predictors of mortality in 156 male alcoholics who were psychiatrically hospitalized between 1967 and 1968 and followed for > 20 years. The findings showed that an older age, liver cirrhosis, and separation or divorce at intake were all associated with a shorter time to death. These results corroborate those of other follow-up studies that have identified age, health, and social support as independent correlates of mortality in alcoholics. From a therapeutic standpoint, the most alterable predictor is social support. Thus, the establishment of a salutary social network and social support system should be an integral part of the alcoholic's treatment plan. PMID- 7485851 TI - EEG dimension in sons of alcoholics. AB - Recent advances in the field of nonlinear dynamics have provided new conceptual models, as well as novel analytical techniques applicable to studies in alcohol research. One measurement technique, which has been developed in an attempt to characterize nonlinear systems in physics and biology, is the estimation of attractor dimension. This dimension may be seen as a measure of the information required to describe the current behavior of a system. We have applied these techniques to the analysis of EEG collected from 17 men with alcoholic fathers and 19 men with no alcoholic relatives. The men with alcoholic fathers were found to have a lower EEG attractor dimension than their matched controls. This suggests that the EEG of young men with alcoholic fathers may be "more organized" or "less complex" than men with no alcoholic relatives. Although more studies will be needed to explore this hypothesis, these studies suggest that further development of nonlinear approaches to the analysis of brain systems are likely to generate new clinical measures, as well as new ways of viewing brain electrical function and alcoholism. PMID- 7485852 TI - Effectiveness of the Alcohol Use Disorders Identification Test in occupational health screenings. AB - This study evaluated the effectiveness of the 10-item Alcohol Use Disorders Identification Test (AUDIT) among occupational health screenings. AUDIT was available from 32 male and 93 female employees, mainly of academic organizations; 22 alcoholic men reporting to treatment at a detoxification clinic served as a reference group. Two other structured questionnaires, the Malmo modified Michigan Alcoholism Screening Test (Mm-MAST) and the CAGE, were compared with the AUDIT, and comparisons were also made to the self-reported weekly alcohol consumption. The total score on the AUDIT correlated equally with the self-reported alcohol consumption, especially when the alcoholics were excluded (r = 0.73, p < 0.001) as the Mm-MAST (r = 0.58, p < 0.001) and CAGE (r = 0.33, p = 0.005). All three questionnaires were good at detecting male alcoholics: using the cut-off point of at least 8 positive answers on the AUDIT, 3 on the Mm-MAST, and 3 on the CAGE. The sensitivities were 100%, 100%, and 91%, respectively. In the AUDIT, however, there was a clearer difference related to the cut-off level than with the Mm-MAST and CAGE. With the cut-off of 8 points, 31% of the male and 11% of the female employees were classified as suspect heavy drinkers. The AUDIT was significantly more often accurate than the two other questionnaires, especially among female employees in detecting suspect early-phase heavy drinkers and thus seems to be suitable for health screenings. PMID- 7485853 TI - [The microbiology of secretory otitis]. AB - The AA. realize a comparative study on the differences between the nasopharyngeal microbial flora of 50 children suffering a secretory otitis and other 40 children without middle ear disease. In nasopharyngeal cultures the pathogenic flora (Haemophilus influenzae, Streptococcus pneumoniae, Moraxella catarrhalis, Streptococcus beta hemoliticus group A, Staphilococcus aureus) amounted for 96 percent in children with secretory otitis, which figure was reduced to 80 percent in healthy infants (p < 0.05). Haemophilus influenzae was the most identified microorganism in a both nasopharyngeal and otic flora. We have found a significative association (p < 0.001) among nasopharyngeal and otic flora of each individual. PMID- 7485854 TI - [The dorsal flap in laboratory animals]. AB - A total of 108 flaps were fashioned in the dorsal area of laboratory animals (47 pedicle and 7 free) and 54 myocutaneous (32 pedicle and 22 free). The anatomical characteristics of the dorsal flap in the rabbit are presented and the microsurgical technique utilized in elaborating free flaps are described. In comparing survival of the different flap types, a greater survival rate (p < 0.05) was found for myocutaneous than cutaneous; and for pedicle than free flaps. In addition it was verified than arterial flow is indispensable for flap survival. PMID- 7485855 TI - [Sudden bilateral deafness as allergic manifestation]. AB - The paper reports the case of a young woman who presented with a serious bilateral sudden deafness, after the ingestion of a betalactam antibiotic (known as allergic for her) in association with a mucolytic. Account of the course of the clinical events noticed, which are compared with the findings observed in other scarce publications on sudden deafness linked with allergic reactions. Finally the AA. outline a pathogenetic hypothesis in order to explain these cases of deafness involved with digestive allergic reactions. PMID- 7485856 TI - [Osseous hemangioma of the upper jaw]. AB - Osseous hemangioma is a rare tumour of head and neck localisation. In this work the AA. present a case of the tumour located in the upper maxilla, considering the actual concept and the possibilities of differential diagnosis in relation with both radiologic and anatomopathological characteristics. PMID- 7485857 TI - [DNA papillomavirus detection in head and neck benign proliferative lesions]. AB - In a retrospective study, human papillomavirus (HPV) (6/11, 16 and 18 types) were tested using polymerase chain reaction (PCR), in stored formalin-fixed, paraffin embedded tissue sections, from 36 head and neck adult papillomata. The results, only 20 percent of positive cases, are non consistent with the role of the HPV infection in the etiology of head and neck papillomata in adult patients. However we detect HPV-18 positivity in papillomata with dysplasia. PMID- 7485858 TI - [Descriptive study of singing voices in children and women]. AB - This paper is focused in the study of singing voices of both women and children, with the purpose of displaying and verify the existing differences of the two. In this article will be made clear than the affirmation made by Husson, Garde and Arribas (1962, 1973 and 1975, respectively) is not a dogmatic statement. The method used by us in order to drawn out the conclusions are the following: 1. A discriminative audio-perceptive technique requiring skilled experts in order to settle if the recorded voices belong to women, boys or girls. 2. Other technique adopted is founded in the spectral analysis of fragments recorded with the integrator of spectral density. The obtained results are as follows: The analysis audio-acoustic disclose differences between feminine voices and children voices. The experts could, with certainty, differentiate between each one of them (women 74%; children 79%). By means of the spectrum analysis was possible to verify that the prime objective of the spectral analysis employed is able to mimicking the global subjective impression received through the human auditory system. PMID- 7485859 TI - [Peripheral facial palsy associated with rhinopharyngeal tumor]. AB - Peripheral facial paralysis from tumoral origin is scarce in number, in spite of possible manifold causative growths. One case in which the palsy was associated to a rhinopharyngeal neoplasm is reported and possible ways of nerve involvement discussed. Although progression seems to be the most probable hypothesis in this instance, only the anatomo-pathological exam has affirmative value. PMID- 7485861 TI - [Nitric oxide and opiate dependence]. PMID- 7485862 TI - [A magnificent Spanish surgeon: Placido Gonzalez Duarte. 1887-1986]. PMID- 7485860 TI - [Multicenter study comparing the efficacy and tolerance of topical ciprofloxacin (0.3%) versus topical gentamicin (0.3%) in the treatment of simple, non cholesteatomaous chronic otitis media in the suppurative phase]. AB - A multicentre double-blind randomized study was carried out to compare topical ciprofloxacin and topical gentamicin in the treatment of simple non cholesteatomatous purulent chronic otitis media. Three hundred and eight patients were included in the study, 159 treated with ciprofloxacin and 149 treated with gentamicin. The percentage of clinical success (elimination of otorrhoea) was 95% with ciprofloxacin and 94% with gentamicin (ns). Likewise, the percentage of bacteriological erradication was 96% with ciprofloxacin and 93% with gentamicin. Both drugs were well tolerated, without changes in the audiometric values. In these patients, topical ciprofloxacin shows the same efficacy as topical gentamicin without any potential ototoxic effect. PMID- 7485863 TI - [Veterinary sciences and their historical affinity with medicine]. PMID- 7485864 TI - [Thalassotherapy: its current therapeutic influence]. PMID- 7485865 TI - [The epiphysis in reproduction (the diapause and the sexual cycle)]. PMID- 7485866 TI - [Vegetarianism in childhood and adolescence]. PMID- 7485867 TI - Euthanasia: the Dutch experience. PMID- 7485868 TI - [Euthanasia: the status of the question]. PMID- 7485869 TI - 'Walking' epidurals in labour. PMID- 7485871 TI - Compliance with an incident report scheme in anaesthesia. AB - Voluntary anonymous reporting of incidents which occur during anaesthesia is a mechanism for obtaining information about such problems. Our objective was to estimate with reporting such incidents. For 3 months, alongside the incident report form, another form, which had to be completed for every patient, was used. This form listed a series of defined events which could occur intra-operatively. Compliance with the incident reports was estimated by comparing the data obtained from the two sets of forms. Overall compliance was only about 30%. There were differences in compliance for different events. Compliance was high with more serious events and poor in the case of common events, or when successful recovery had occurred. In order to improve compliance, incident report forms should be available on each anaesthetic machine and staff should be made more aware of the fact that reportable incidents are not limited to events which harmed the patient, but also include those which could have affected patient safety. PMID- 7485870 TI - Molecular sieves: an alternative method of carbon dioxide removal which does not generate compound A during simulated low-flow sevoflurane anaesthesia. AB - Molecular sieves are used in industry to 'scrub' industrial gases. We examined, during simulated low-flow closed system anaesthesia, (1) the carbon dioxide adsorbing potential of molecular sieves and (2) the reactivity of the sieves compared to soda lime using sevoflurane as an indicator. A low-flow anaesthetic system containing 13X molecular sieves was connected to a model lung. End-tidal concentrations of CO2 were measured continuously at an O2 flow of 800 ml.min-1 and a CO2 flow of 200 ml.min-1. In the second study, sevoflurane (FE'sevo 1.7%) was added to the system after which samples were taken from the inspiratory limb of the anaesthetic system. This experiment was performed both during carbon dioxide removal with soda lime and with the molecular sieves. The samples were stored in gas-tight syringes and analysed by gas chromatography. The temperature of both absorbents was measured throughout the study. The molecular sieves adsorbed carbon dioxide (20%) efficiently for a period of 5 h. There was a gradual increase from the baseline of 4.4% to 4.5, 5.4, and 6.0% at 90, 180, and 300 min, respectively. When sevoflurane was added to the system, compound A was detected at the start of both experiments. However, when soda lime was used the concentrations of compound A increased 10-fold after 2.5 h compared with baseline values. No increase in compound A was observed when molecular sieves were used for carbon dioxide removal. The highest mean (SD) temperature of the molecular sieves was 41.5 (3.2) degrees C. Molecular sieves are effective adsorbents of carbon dioxide when used in a simulated low-flow, closed anaesthetic system.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7485873 TI - Variable-dose patient-controlled analgesia. A preliminary report. AB - One size of patient-controlled analgesia demand dose does not suit all patients after surgery. We have constructed a new patient-controlled analgesia (variable dose patient-controlled analgesia) in which patients have a choice of demand dose sizes. In an initial trial, patients could choose between 0.5, 1.0 or 1.5 mg morphine. Patients readily understood the system and were all 'satisfied' (n = 1) or 'very satisfied' (n = 9) with the system. Only two patients sought 'complete relief' from pain, most reported seeking 'moderate relief'. All patients obtained a 'lot of relief' or 'complete relief' with this patient-controlled analgesia system. This preliminary experience suggests that variable dose patient controlled analgesia warrants further investigation and comparison with conventional patient-controlled analgesia. PMID- 7485872 TI - Lignocaine metabolite formation: an indicator for liver dysfunction and predictor of survival in surgical intensive care patients. AB - Formation of the lignocaine metabolite monoethyl-glycine-xylidide (MEGX) by hepatic cytochrome P450 enzymes is a new method for evaluating liver function. The purpose of this study was to compare MEGX formation with other liver function parameters in surgical intensive care unit patients. The study include 29 critically ill patients who had been admitted to the unit for more than 3 days with a median APACHE III score-predicted mortality > 30%. On day 4, lignocaine was given intravenously at a dose of 1 mg.kg-1 over 2 min and MEGX formation was measured 15 min later. Eighty-nine percent of the patients had MEGX values below 90 micrograms.l-1 indicating impaired liver function. Eleven patients died, 18 patients survived. The group of patients with fatal outcome had significantly lower MEGX values (median: 23 micrograms.l-1) than the group of survivors (median: 53 micrograms.l-1, p < 0.01). Bilirubin values were elevated in the non survivor group (median: 2.8 mg.dl-1) compared to the survivors (median: 0.9 mg.dl 1, p < 0.05). There was no significant difference between two groups in the other liver function tests. We conclude from our results that the MEGX test can be considered an indicator for hepatic dysfunction and predictor of survival in critically ill patients. PMID- 7485874 TI - The quality of breathing and capnography during laryngeal mask and facemask ventilation. AB - Clinical measures of ventilation and the relationship between arterial and end tidal carbon dioxide tensions were studied during inhalational anaesthesia in 18 patients using a laryngeal mask airway or a facemask. Tidal volumes were similar in both groups but expired minute volume, respiratory rate and physiological deadspace ventilation were significantly increased in the facemask group. Both end-tidal and arterial carbon dioxide tensions were higher in the laryngeal mask group. Mean arterial to end-tidal carbon dioxide tension differences ranged from 0.13 to 4.13 kPa in the facemask group and from 0-1.73 kPa with the laryngeal mask airway. Pooled data analysis revealed a better correlation between arterial and end-tidal carbon dioxide tensions during laryngeal mask ventilation as compared to facemask breathing. With both techniques the arterial to end-tidal carbon dioxide tension difference was related to respiratory rate and physiological deadspace ventilation. Estimation of arterial carbon dioxide partial pressure by monitoring end-tidal carbon dioxide tension is more reliable with the laryngeal mask airway than during facemask breathing, in particular at small tidal volumes. PMID- 7485875 TI - A cadaver study appraising accuracy of blind placement of percutaneous tracheostomy. AB - An attempt was made to place a percutaneous catheter between the first and second cartilaginous rings of the trachea in 20 cadavers presenting for postmortem examination. Only nine of the catheters entered the trachea in the intended space. All catheters which were noted to be difficult to place entered the trachea higher than intended. One third of all catheters punctured the thyroid isthmus. PMID- 7485876 TI - The use of the Hayek Oscillator during microlaryngeal surgery. AB - An extremely mounted cuirass ventilator, the Hayek Oscillator, was used on 41 patients undergoing surgery to the larynx without the use of tracheal tube. Gas exchange and cardiovascular parameters remained satisfactory during the use of this technique, which offers a significant advance over existing tubeless methods of anaesthesia. PMID- 7485877 TI - The Ohmeda Universal PAC drawover apparatus. A technical and clinical evaluation. AB - The Ohmeda Universal PAC drawover apparatus, a modification of an earlier series of vapour-specific vaporizers, was assessed with regard to its output under varying gas flows and temperatures and its clinical usage with isoflurane and enflurane. The device tended to over-deliver vapour, particularly at low flows and at high temperatures. Its clinical performance during spontaneous and positive pressure ventilation was satisfactory, although adequate concentrations were not achieved using enflurane in the spontaneous ventilation mode. The device was not assessed with ether or halothane. The robust construction, relatively large capacity and thermocompensation make it suitable for field or military anaesthesia. PMID- 7485879 TI - Exponentials and the anaesthetist. 1964. PMID- 7485880 TI - Cardiopulmonary complications during anaesthesia and surgery for severe thoracic lordoscoliosis. AB - A 15-year old girl underwent a posterior spinal fusion procedure for severe lordoscoliosis. On turning to the prone position acute posture-related upper airway obstruction occurred. During surgery, manipulation of the spine caused several episodes of loss of cardiac output. Retrospective assessment of the pre operative chest X rays demonstrated approximation of the sternum and thoracic vertebrae, with evidence of tracheal narrowing at the level of the thoracic inlet. We feel that both complications were directly related to the severity of the lordoscoliosis and that careful pre-operative clinical, spirometric and X ray assessment should be undertaken in such patients. PMID- 7485882 TI - Skin-epidural distance in children. AB - The skin-epidural distance was measured in 274 children to assess the usefulness of 1 mm.kg-1 as a guideline. Children aged between 2 days and 16 years, weighing between 2 and 43 kg were investigated. Lumbar epidurals were performed under general anaesthesia using a midline approach in the L3-4 interspace with the patient in the lateral position. Good correlation between skin-epidural distance and age and weight was demonstrated. No statistical difference was shown when plotting the best fit regression line: skin-epidural distance (mm) = 0.8 weight (kg) + 3.93 (R2 = 0.74) and its 95% confidence limits and skin-epidural distance (mm) = weight (kg) for children between 6 months and 10 years. Poor correlation was noted below 6 months (n = 22) and over 10 years (n = 19). No dural puncture or bloody tap occurred. One mm.kg-1 body weight was shown to be a useful guideline for children between 6 months and 10 years of age. PMID- 7485878 TI - A microprocessor based photoplethysmograph for use in clinical practice. AB - A microprocessor-based system of photoplethysmography and its use in measuring skin blood flow is described. The system was designed specifically for use over long periods and for the analysis of the output to be handled by a personal computer. The photoplethysmograph described in this paper is shown to be a qualitative method for assessment of changes in peripheral skin blood flow. There is potential for use of this instrument in a variety of clinical conditions. PMID- 7485881 TI - Skin tethering after dilatational percutaneous tracheostomy. AB - Percutaneous tracheostomy is increasingly preferred for patients in the Intensive Care Unit. It is a convenient method with a low incidence of complications. Skin tethering has not previously been emphasized as a complication of this technique and, although primarily cosmetic, may result in significant morbidity. PMID- 7485883 TI - Review of uses of the laryngeal mask in ENT anaesthesia. AB - A review of the various uses of the laryngeal mask in otorhinolaryngological surgery is presented. Its primary use is in airway management, especially during recovery from anaesthesia. PMID- 7485884 TI - Post cholecystectomy admission to the intensive care unit. Comparison between open, mini-lap and laparoscopic techniques. AB - Admissions to the intensive care unit following techniques for cholecystectomy, the open, mini-lap and laparoscopic methods, were studied over an 18-month period. Of 1078 cases of operative cholecystectomy performed during that period, 28 cases (2.6%) from all groups were admitted to the intensive care unit. Cases in the open and mini-lap cholecystectomy groups were admitted for postoperative monitoring because of pre-existing medical diseases and were discharged after an average period of stay of 2.4 and 4.7 days respectively. In contrast, the average stay of patients in the laparoscopic group was 14.8 days and the majority were admitted as a result of complications related to the procedure. In this group, one patient died on the first day of admission to the unit (mortality of 8.3%). This study has shown that laparoscopic surgery, from the intensive care point of view, is a very costly surgical procedure. PMID- 7485886 TI - The effect of cerebrospinal fluid dilution of isobaric 0.5% bupivacaine used for spinal anaesthesia. AB - A prospective study was conducted to see the effect on spinal anaesthesia of the dilution of isobaric 0.5% bupivacaine with cerebrospinal fluid. Sixty patients were randomly allocated to three groups. In group 1, patients received 3 ml isobaric 0.5% bupivacaine intrathecally without aspirating cerebrospinal fluid. In groups 2 and 3, cerebrospinal fluid 1 ml and 2 ml was aspirated respectively and mixed with 3 ml isobaric 0.5% bupivacaine. A total volume of 4 ml in group 2 and 5 ml in group 3 was administered. Thus, the volume of cerebrospinal fluid remained unchanged. Pinprick analgesia and motor block was evaluated from induction until recovery. No differences in onset time, duration and 'two segments regression' were noticed. The only statistical difference was the time to reach complete motor block, which was shorter in group 1 as compared to groups 2 and 3 (6.9 SD 1.4 min versus 11.3 SD 3.0 and 13.5 SD 3.9 min respectively). The mean value of maximum decrease in systolic blood pressure was small, being less than 15% of the pre-operative value for each group. In conclusion, the effect of diluting isobaric 0.5% bupivacaine with cerebrospinal fluid, 1 ml and 2 ml, is minimal and it is an unnecessary procedure with limited clinical effect. PMID- 7485887 TI - Endangered opportunities in acute pain management. PMID- 7485885 TI - A comparison of diclofenac with ketorolac for pain relief after knee arthroscopy. AB - We performed a double-blind controlled trial to compare the analgesic effect of two nonsteroidal anti-inflammatory drugs. We compared rectal diclofenac 100 mg given 1 h before induction of anaesthesia with intravenous ketorolac 10 mg given immediately before anaesthesia in 40 patients undergoing arthroscopy of the knee as day cases. A visual analogue scale was used to assess pain prior to discharge. Pain, analgesic consumption, sleep disturbance and restriction of activities were recorded by telephone enquiry 24 h after surgery. There was no difference in the pain parameters, sleep disturbance, or restriction of activity between groups. We suggest that ketorolac 10 mg intravenously and diclofenac 100 mg rectally provide comparable postoperative analgesia in the first 24 h after arthroscopy of the knee. PMID- 7485888 TI - Dangers of CO2 cylinders on anesthetic machines. PMID- 7485889 TI - Cricoid pressure and advance life support. PMID- 7485893 TI - The seven year itch. PMID- 7485892 TI - Measuring PCO2 for gastric tonometry: precision is as important as bias. PMID- 7485891 TI - Cricoid pressure and advance life support. PMID- 7485890 TI - Cricoid pressure and advance life support. PMID- 7485894 TI - A trainer for identification of the epidural space. PMID- 7485895 TI - Peribulbar anaesthesia using the immobile needle technique. PMID- 7485896 TI - HME filter and Ayre's T piece. PMID- 7485897 TI - Does propofol cause deaths in children? PMID- 7485898 TI - Kinking of a tracheal tube in the nasal cavity. PMID- 7485899 TI - Bite block for the laryngeal mask airway. PMID- 7485900 TI - Safety not speed. PMID- 7485901 TI - Upper and lower limb blood pressures during general anaesthesia. PMID- 7485903 TI - Precipitation in PCA mixtures. PMID- 7485904 TI - A hidden leak. PMID- 7485902 TI - Overdose of morphine during PCA. PMID- 7485905 TI - One-way valve malfunction in a circle system. PMID- 7485907 TI - Maternal position during induction of spinal anaesthesia for caesarean section. PMID- 7485906 TI - Induction end-point and laryngeal mask placement. PMID- 7485908 TI - Anaesthetic techniques for chair dental anaesthesia. PMID- 7485909 TI - Recognition of the anaesthetists: a Spanish view. PMID- 7485910 TI - Is there a need for a new inhalational anaesthetic agent? AB - New anaesthetic agents are continually being developed in an effort to prepare the ideal agent. Isoflurane is the first choice of many anaesthetists, although it clearly falls short of the ideal on a number of counts. The need and place for any new agent depends on comparing the properties of a theoretically ideal agent with those of isoflurane and the new agent. Desflurane is a step towards a more ideal inhaled agent; however, there is still the need to synthesise and evaluate new chemical entities in order that the ideal can more nearly be approached. PMID- 7485911 TI - Cardiovascular effects of desflurane in experimental animals and volunteers. AB - Desflurane (difluoromethyl-1-fluoro-2,2,2-trifluoroethyl ether [I-653]) was introduced into clinical practice in 1992. The present review summarises the current state of knowledge of the cardiovascular effects of desflurane, with emphasis on investigations published in the past 2 years, and compares desflurane's effects with those of isoflurane, and where available, sevoflurane. Desflurane produces two distinct cardiovascular actions. First, desflurane decreases left ventricular systolic and diastolic function to an extent similar to that of isoflurane. It decreases systemic vascular resistance and mean arterial blood pressure in a dose-dependent fashion. Heart rate is unchanged at lower steady-state concentrations, but increases with higher concentrations. Addition of nitrous oxide maintains heart rate unchanged, increases systemic vascular resistance and mean arterial blood pressure, but decreases cardiac output. Secondly, when the end-tidal concentration of desflurane is very rapidly increased to concentrations exceeding 1 MAC, in the absence of premedication, desflurane increases sympathetic activity, heart rate and mean arterial blood pressure. Prior administration of fentanyl, esmolol or clonidine blunts this response. The response and the ability of fentanyl to blunt it probably account for the disparate results in published studies on the use of desflurane in patients undergoing coronary artery bypass graft surgery. PMID- 7485913 TI - Recovery from outpatient anaesthesia. Factors affecting outcome. AB - In 1994, 66% of all surgery in the USA was performed as ambulatory surgery. Day surgery is also expanding to other countries worldwide. To provide safe anaesthesia and good outcomes for longer and more extensive operations performed in ambulatory facilities, patients must be carefully evaluated before surgery, their home readiness must be assessed, and they must fully understand all relevant information. Good outcome requires adoption of policies for safe discharge from the hospital. If a patient does not have an escort home, the surgical procedure should be cancelled or the patient admitted to the hospital. As the number of patients and complexity of scheduled surgical procedures increases, the outcome of day surgery will increasingly depend on the anaesthetist's skills. The recently introduced short-acting drugs may further improve the outcome after day surgery by facilitating rapid recovery and an early return to normal daily activities. PMID- 7485912 TI - Circulatory effects of desflurane. AB - The cardiovascular properties of desflurane and some other inhalational anaesthetics are outlined and compared, and the way in which they may be effectively used in clinical practice is considered. Desflurane appears to offer some advantages compared with currently available anaesthetics. Its effects on left ventricular function are moderate and the solubility of the agent in blood and tissue is remarkably less than isoflurane, enflurane, or halothane. PMID- 7485914 TI - Inhalation anaesthesia at the extremes of age: geriatric anaesthesia. AB - No single anaesthetic technique is superior for all elderly patients, although the prognosis is improved if minor surgical procedures are performed with local anaesthesia rather than with general or major regional anaesthesia. If general anaesthesia is required, which specific inhalation anaesthetic can improve the prognosis of the geriatric patient? The low solubility of desflurane and its resistance to biodegradation would seem to recommend its use in the geriatric patient. However, these theoretical advantages remain to be adequately documented. PMID- 7485915 TI - Physicochemical properties and pharmacodynamics of desflurane. AB - Desflurane is a new volatile, inhaled anaesthetic that differs significantly from presently available inhaled agents in being halogenated solely with fluorine. This fluorination produces a lower solubility and increased resistance to biodegradation. The lower solubility provides a more precise control over, and a more rapid awakening from, anaesthesia. The resistance to biodegradation suggests that toxicity from desflurane may be minimal or non-existent. Limitations include a lesser potency and greater pungency at concentrations exceeding 1 minimum alveolar concentration (MAC). Other pharmacological properties are similar to those of isoflurane. PMID- 7485917 TI - Low-flow anaesthesia. AB - Although many anaesthesia machines are equipped with circle rebreathing systems, inhalational anaesthesia remains frequently performed using relatively high fresh gas flows. The major advantages of rebreathing techniques can be achieved only if the fresh-gas flow is reduced to 1 l.min-1 or less. Although there are potential risks associated with low-flow anaesthesia, modern anaesthesia machines meet all the technical requirements for the safe use of low-flow techniques if they are used in conjunction with equipment for monitoring inhaled and exhaled gas concentrations; these monitors are already increasingly available and, in the near future, are likely to become an obligatory safety standard in many countries. For both economic and ecological reasons, the use of new inhalational anaesthetics, with low tissue solubility and low anaesthetic potency, can be justified only if the efficiency of administration is optimised by using low-flow anaesthetic techniques. PMID- 7485916 TI - Inhalational anaesthesia at the extremes of age: paediatric anaesthesia. AB - The relevant physiological and pharmacokinetic differences between adults and infants and children are considered, and the advantages and disadvantages of the available inhalational anaesthetics are discussed. Desflurane shows promise as a new paediatric anaesthetic as it allows rapid changes in the depth of anaesthesia. However, irritant effects on the airways make desflurane unsuitable for induction. Desflurane anaesthesia, following halothane induction, seems to be well tolerated. PMID- 7485918 TI - Economic analysis and pharmaceutical policy: a consideration of the economics of the use of desflurane. AB - Several factors have to be considered in determining the cost of applying a new inhalational anaesthetic such as desflurane into clinical practice. Factors beyond the immediate control of the anaesthetic practitioner include the price set by the manufacturer (although this may be influenced by economic and political pressures), and the physical-pharmacological properties of the anaesthetic (e.g. vaporization, potency, solubility). The anaesthetic practitioner can minimise cost by applying lower inflow rates. Lower solubility (and hence lower uptake) provides a greater economy at lower inflow rates than does higher solubility. Furthermore, lower solubility permits the use of lower inflow rates with greater precision to the control of anaesthesia, and greater ease of application. At present unit prices, the cost of desflurane approximately equals that of isoflurane when a 1 l.min-1 inflow rate is used. The use of lower inflow rates presupposes that such rates do not allow the production of toxic compounds in recirculating gases. Modern equipment makes low-flow anaesthesia reliable and easy to control, and as desflurane is not degraded by the standard carbon dioxide absorbents, its use in low-flow systems is effective and economical. These cost considerations do not take into account the savings that may result from a more rapid recovery from anaesthesia, nor do they take into account the increased expense of capital equipment needed to apply a new anaesthetic. PMID- 7485919 TI - Economic analysis and pharmaceutical policy. AB - Economic evaluation, a comparative analysis of alternative actions in terms of costs and consequences, allows rational decisions to be made concerning the deployment of resources (people, time, equipment, facilities and knowledge). Pharmaceutical policy reflects the various objectives of the many social groups, some of which are conflicting. While new methodologies for evaluation of health care programmes still need to gain wider acceptance, resource limitations for both care providers and decision makers make economic analysis an increasingly important tool. PMID- 7485921 TI - [Therapy of acute coronary insufficiency with phosphodiesterase III inhibitors. Symposium. Hamburg, 13 February 1993]. PMID- 7485920 TI - Implications of chemical and physical properties of desflurane for longer surgery. AB - Increased duration of anaesthetic administration has implications for recovery from anaesthesia, has cardiovascular effects, and potential for toxicity through metabolism and breakdown of the anaesthetics. Recovery of function after desflurane or sevoflurane anaesthesia, because of the low blood gas and tissue solubilities of these agents, is more rapid than after halothane, isoflurane or enflurane, with recovery being most rapid after desflurane. Increased duration of anaesthesia amplifies the differences in rate of recovery because of the additional anaesthetic (greater with more soluble agents) dissolved in tissues. Increased duration of anaesthesia lessens the cardiovascular depression associated with most halogenated inhaled anaesthetics including desflurane, but not isoflurane. Increased duration of anaesthesia allows for greater metabolism of anaesthetics and greater exposure to metabolites and potentially toxic breakdown products. Desflurane is the least metabolised of the available anaesthetics and is stable in soda lime and Baralyme. Thus, it has exceedingly low potential for toxicity. Sevoflurane undergoes considerable metabolism, producing free fluoride ion, with plasma concentrations proportional to dose and duration of anaesthesia exceeding 50 microM in approximately 7% of patients. In rats, the effects of a toxic breakdown product of sevoflurane, CF2 = C(CF3)OCH2F (compound A), are also dose- and duration-dependent, with lower concentrations producing toxic effects as duration of exposure increases. The clinical importance of the metabolism and in vitro breakdown of sevoflurane has still to be adequately tested. PMID- 7485922 TI - [Preclinical control of intubation and artificial respiration. Animal experiment and literature review]. AB - Oesophageal malposition of an endotracheal tube is among the leading causes of anaesthesia incidents. While clinical manoeuvres for detection of tube malposition are unreliable, monitoring (i.e. capnography) can prevent such incidents. The problem is particularly important in prehospital care, where capnography is not (yet) widely available. We tested three devices used for differentiating oesophageal from endotracheal intubation: 1. Non-CO2-dependent Oesophageal Detector Device (ODD) as described by Pollard and Wee, 2. Semi quantitative chemical disposable capnometer EasyCAP (Nellcor), 3. Non quantitative infrared miniaturised capnometer MiniCAP (MSA). METHODS. 50 anaesthetised minipigs were intubated with a Magill tube. An identical additional tube was placed in the oesophagus. The cuffs of both tubes were inflated. Unexperienced personnel (students, laboratory technicians) were asked to determine the position of one of the tubes by using one of the devices according to the randomisation plan. The decision had to be taken within 30 s. Using the ODD, the proband first injected 100 ml air into the lung (or stomach) and then tried to aspirate the same volume. EasyCAP and MiniCAP were used according to manuals. RESULTS. Each device was used 25 times with a tracheal tube and 25 times with an oesophageal tube. All tube position identifications were correct. When ventilating the oesophagus/stomach for capnometric control, regurgitation into the tube occurred six times (five times with the EasyCAP and once with the MiniCAP). In these cases, the decision was based on this occurrence and not on the display of the device. While using the ODD no regurgitation occurred. CONCLUSION. These devices are useful for preclinical practice. According to the literature and our experience, the ODD is superior for the initial control of tube position, especially in cardiac arrest. Capnometry is needed, however, for continuous control of ventilation. PMID- 7485923 TI - [Erythropoietin therapy during frequent autologous blood donations. Dose-finding study]. AB - Avoidance of homologous blood products and patients' demand for preoperative autologous blood donation programs are increasing. As many of these patients are older, with a compromised cardiovascular system and a slow response of the erythropoietic system when anemia occurs, the feasibility and benefit of autologous blood donation is often limited. Augmentation of preoperative blood donation by therapy with recombinant human erythropoietin (rHuEPO) has been described in animal models and in patients. METHODS. In a multicenter, controlled, randomized trial, 49 patients scheduled for orthopaedic or vascular surgery received 0 (control group, n = 9), 200 (n = 10), 300 (n = 11), 400 (n = 10) or 500 (n = 9) U/kg rHuEPO (Erypo, Cilag, Sulzbach, distributor Fresenius, Oberursel, Germany) subcutaneously twice a week for 3 weeks while every week 450 ml blood was collected. Iron sulphate 100 mg was prescribed orally twice a day. Patients were ineligible if they had uncontrolled hypertension, recent myocardial infarction, haematological disorders or a history of seizures. Blood donation had to be cancelled if the haematocrit was below 30%. RESULTS. There was a significant (ANOVA) drop of the haematocrit value only in the control group, and end-point values for haematocrit and haemoglobin were significantly elevated in the 400 and 500 U/kg groups compared with the control group (Table 9). DISCUSSION. The erythropoietic stimulus of phlebotomy for autologous blood donations is often not efficient enough to guarantee a constant haematocrit. Lowering of the preoperative haematocrit jeopardizes the aim of avoidance of homologous blood transfusions. rHuEPO increased the efficiency of autologous blood collections, as predonation haematocrit values could be preserved in the high-dosage groups. As a consequence, homologous transfusions could be avoided. However, there were broad interindividual differences in the erythropoietic response, possibly due to limitations in iron availability. Adverse effects of rHuEPO therapy, such as hypertension, thrombosis or neurologic disorders, are mostly reported in patients with terminal kidney failure. No such disturbances were observed in the present study. CONCLUSION. rHuEPO ameliorates the preoperative decrease of haemoglobin and haematocrit values due to autologous blood donations in a dose-related fashion. The individually adjusted dosage of rHuEPO and iron supplementation merits further investigation. PMID- 7485924 TI - [Autologous blood donation in obstetrics]. AB - OBJECTIVE: To demonstrate the safety of autologous blood donation in an obstetric practice. METHODS: A total of 2350 pregnant patients were screened for an increased risk of peripartum bleeding. Excluding patients with contraindications, a total of 71 women participated in this autologous blood donation program. Blood was donated in the 35th and 37th weeks of pregnancy. RESULTS: No problems occurred in the course of blood donation. All newborns were found to be healthy; 41 of the 71 patients received peripartum autologous blood transfusions. Homologous blood was not given to any of the patients. CONCLUSION: Our experience demonstrates that autologous blood donation is a safe practice in the setting of obstetrics, when the indications and contraindications are considered. Further studies in larger patient populations are necessary to expand or restrict the indications for autologous transfusion in obstetrics. PMID- 7485925 TI - [Multidimensional psychometric assessment of preoperative mood. Effects of zolpidem compared to phenobarbital combined with promethazine as premedication]. AB - The following double-blind, randomised study dealt with three questions: (1) Is a multidimensional psychometric rating scale suitable for the measurement of mood before anaesthesia? (2) What are the effects of the new benzodiazepine-like drug zolpidem on preoperative mood compared with phenobarbital? (3) Is the combination with Promethazine suggestive? METHODS. Three hundred and four patients were assigned to four groups (group 1: zolpidem 8.03 mg/promethazine 50 mg; group 2: zolpidem 8.03 mg/placebo; group 3: phenobarbital 100 mg/promethazine 50 mg; group 4: phenobarbital 100 mg/placebo). The drugs were given the evening before anaesthesia (09:30-10:00 p.m.). The sample was shifted by age and sex. Mood was measured by a multidimensional rating scale, which assessed aspects of elated mood, anxiety, hostility, deactivation, vigilance, and introversion. Statistics were performed using analysis of variance (ANOVA). RESULTS. Zolpidem led to significantly higher expressions of hostility (negative mood, irritability, aggressiveness) than phenobarbital. Compared with placebo, promethazine led to greater deactivation (more tiredness and numbness, lower level of wakefulness). Specific emotions and somatic aspects were not affected. Patients who had received promethazine received a lower dose of thiopentone for induction of anaesthesia than patients with placebo. CONCLUSIONS. Zolpidem and phenobarbital have many common effects on preoperative mood. Differences were found in the unspecific emotional aspects of agitation and hostility. These negative effects must be weighed against the pharmacokinetic and pharmacodynamic advantages of zolpidem when this drug is administered for premedication. The effects of zolpidem seem to be more sedative than anxiolytic. The study shows that a combination with promethazine is suggestive, because promethazine has a selective deactivating effect. The finding that promethazine lowers the dose of thiopentone required for induction of anaesthesia is an additional interesting point. The results of this study highlight the importance of using multidimensional rating scales for the measurement of mood before anaesthesia. PMID- 7485926 TI - [Efficiency of a mobile oxygen concentrator for mechanical ventilation in anesthesia. Studies with a metabolic lung model and early clinical results]. AB - Oxygen (O2) for clinical application is generally provided from either a central gas supply via a hospital pipeline system or is delivered to the working place in cylinders as compressed gas. An alternative source is the one-site generation of O2 from air using O2 concentrators based on molecular sieve technology. Whereas O2 concentrators for anaesthesia in remote areas or underdeveloped countries are wide-spread, in Germany their use is common in neither hospitals nor anaesthesiological practice. The maximum O2 content produced by concentrators is 96% with about 4% argon (Ar) and minimal amounts of nitrogen and other noble gases. The total O2 production is systematically limited, and therefore, the delivered concentration decreases with higher flows. There is also a potential possibility of Ar accumulation in rebreathing anaesthesia systems with reduced fresh gas flow. We investigated the efficiency and potential disadvantages of using O2 concentrators in anaesthesia and the influence of Ar on the accuracy of anaesthetic gas monitors. METHODS. The efficiency of the concentrator was characterised as O2 concentration depending on delivered gas flow. The degree of Ar accumulation in rebreathing anaesthesia systems was obtained with an O2 consuming and CO2-producing metabolic lung model consisting of a water-cooled burning chamber with an adjustable gas jet. The expiratory CO2 content was set to approximately 7%, representing an O2 consumption of 350 ml/min while ventilating the model with 500 ml tidal volume and 10 breaths/min. The inspiratory O2 concentration was adjusted to 35% or 70%; the fresh gas flow was set to 0.5 or 1 l/min. The accuracy of different types of anaesthesia monitors for O2, CO2, volatile anaesthetics, and nitrous oxide in the presence of Ar was checked in comparison with data obtained with a mass spectrometer. To evaluate the usefulness of O2 concentrators for anaesthetic practice, the function of a respirator-concentrator unit was investigated in clinical routine for 8 weeks. RESULTS. The efficiency of the concentrator is flow-rate dependent: O2 concentrations higher than 90% are only achieved with flow rates below 5 l/min and decrease to values lower than 50% at 12 l/min or more. Ar accumulation occurred in rebreathing circuits but exceeded values higher than 10% only under minimal-flow conditions (fresh gas flow 0.5 l/min). Ar did not influence the accuracy of common anaesthetic gas monitors. In clinical practice, the performance of anaesthesia using O2 from an O2 concentrator generated no additional problems. CONCLUSIONS. For the future, the use of O2 concentrators for anaesthesia seems to be a practicable alternative to compressed O2 from cylinders. The main application could be in small operating units or anaesthesia practices. The method is safe and without additional risk of hypoxia, even in rebreathing systems and closed circuits, when the O2 concentration in the inspired gas is measured. PMID- 7485927 TI - [The application of n-acetylcysteine as an antioxidant and mucolytic in mechanical ventilation in intensive care patients. A prospective, randomized, placebo-controlled, double-blind study]. AB - Oxygen radicals and oxygen radial mediators are thought to be important components in the development of acute lung injury, sepsis, and multiple organ failure. Injured patients, patients with pulmonary diseases, and multiple trauma patients also showed an elevated lipid peroxidation, indicating increased oxidant stress. N-Acetylcysteine (NAC) has been used as an antioxidant in a wide variety of experiments. NAC has been suggested to act by raising concentrations of cysteine, and hence glutathione, and by scavenging of oxidant species [1, 11, 17, 29]. The present study was designed to investigate whether the application of NAC in intubated patients has an effect on concentrations of reduced glutathione in plasma and bronchoalveolar lavage fluid (BAL) and on the lipid peroxidation products malondialdehyde and conjugated dienes. Because NAC has been widely used as a mucolytic drug for the treatment of lung diseases, the influence on tracheobronchial mucus was studied, too. METHODS. In a randomized, double-blind, placebo-controlled study, a total of 38 long-term ventilated patients of a surgical intensive care unit were investigated. Patients were treated for 5 days with either 3 g NAC/day or placebo. The plasma concentration of reduced glutathione, malondialdehyde, and conjugated dienes were measured on admission and on the 3rd and 5th days of treatment [8, 34, 48]. Additionally, the numbers of tracheobronchial suctionings were registered and chest radiographs were evaluated. A fibre-bronchoscopy was performed on admission and on the 3rd day of treatment. The amount and viscidity of tracheobronchial secretions were examined semiquantitatively, and glutathione levels were measured in the unconcentrated BAL. The study was approved by the ethics committee of the University of Ulm. RESULTS. The two groups were comparable with respect to age, sex, APACHE II score and diagnosis (Table 1). We found no significant differences in reduced glutathione levels in the plasma or in the BAL (Figs. 1, 2). Plasma concentrations of malondialdehyde were similar (Fig. 3). Only the levels of conjugated dienes were significantly higher on the 5th treatment day in the placebo group (Fig. 4). The organ function of the lung (FiO2, PEEP, PaO2), liver (SGOT, bilirubin), and kidney (creatinine) and coagulation parameters (PTT, prothrombin time, platelet count) were similar in the two groups during the time of investigation. We observed no clinically relevant differences in the tracheobronchial mucus (Table 2). CONCLUSION. The present data do not support routine use of NAC in ventilated patients, either as an antioxidant or as a mucolytic agent. Intravenous administration of 3 g NAC/day had no clinically relevant effect on glutathione levels, lipid peroxidation products, tracheobronchial mucus, and clinical condition. PMID- 7485928 TI - [Operative medicine in the year 2000. 38th Kasseler Symposium, June 23-24, l995]. PMID- 7485929 TI - [The fundamentals of pharmacokinetics]. PMID- 7485930 TI - The Schimmelbusch mask. PMID- 7485931 TI - A review of anatomical and immunological links between epidural morphine and herpes simplex labialis in obstetric patients. AB - The anatomical basis for facial itch after epidural morphone is outlined. CNS nuclear events which reactivate latent herpes simplex and immune inhibition resulting in maternal mouth vesicles or neonatal infections are described. Morphine is hypothesized to affect these processes and facial itch is only a marker, not a trigger of this trigeminal opioid activity. PMID- 7485932 TI - Herpes simplex labialis after epidural or parenteral morphine: a randomized prospective trial in an Australian obstetric population. AB - A prospective randomized controlled trial was conducted in women undergoing emergency and elective caesarean section to determine whether epidural morphine predisposes to postoperative herpes simplex labialis and whether the risk is increased or decreased by previous oral herpes or pruritus. Following cannulation of the epidural space, analgesia for labour and/or caesarean section was induced in two groups of similar age, parity, preoperative status and anaesthetic. According to tables of random numbers, 386 were allocated to receive parenteral morphine and 425 to receive epidural morphine. Postoperative herpes labialis was diagnosed by laboratory culture and microscopy. Patients who received epidural morphine were 11.5 times (95% CI 2.6-49.4) more likely than the parenteral morphine group to suffer herpes labialis. A past history of oral herpes increased the risk of herpes labialis by 8.6 times (95% CI 3.3-22.0). When the influence of past herpes infection and route of morphine were taken into account, no separate association between pruritus and herpes simplex labialis could be demonstrated. PMID- 7485934 TI - Pain and vomiting after vitreoretinal surgery: a potential role for local anaesthesia. AB - Periconal local anaesthesia with subtenon supplementation was used to provide anaesthesia for 94 patients having vitreoretinal surgery. Of these, 44 patients also received general anaesthesia with neuromuscular block. None of these patients received opioid or antiemetic before or during surgery. In comparison with a retrospective control group, patients who had received local anaesthesia as part of their anaesthetic technique were less likely to be given a parenteral opioid (P < 0.001) or to vomit (P < 0.001) within six hours of the completion of surgery. They also experienced significantly fewer bradycardic episodes during surgery (P = 0.001). For patients having general anaesthesia, administration of an intraoperative antiemetic reduced the incidence of vomiting within six hours of the completion of surgery (P = 0.008). For patients who did not receive local anaesthetic, shorter operating time was a factor associated with both reduced postoperative vomiting (P = 0.0015) and administration of parenteral opioid (P = 0.0014). It is suggested that the use of local anaesthesia as part of the anaesthetic technique for vitreoretinal surgery is associated with improved patient comfort. PMID- 7485933 TI - A double-blind randomized prospective study comparing ondansetron with droperidol in the prevention of emesis following strabismus surgery. AB - A prospective double-blind study was conducted to compare the anti-emetic efficacy of ondansetron and droperidol in preventing postoperative emesis following strabismus surgery. A sample size of 213 patients was divided into three equal groups to receive ondansetron 150 micrograms/kg (Group A), ondansetron 75 micrograms/kg (Group B), or droperidol 75 micrograms/kg (Group C). All patients received a standardized anaesthetic technique. All episodes of emesis, recovery time, and time to tolerating oral fluids were recorded. The incidence of emesis during 24 hours was Groups A and B 19.7%, and Group C 28.2%. The lower incidence of emesis recorded by the ondansetron groups compared with the droperidol group was not statistically significant. Ondansetron at 75 micrograms/kg was as effective as 150 micrograms/kg in reducing emesis when compared with droperidol. Mean time to discharge from the recovery room was 75.3 minutes (Group A), 44.4 minutes (Group B), and 41.0 minutes (Group C). The mean time to tolerating oral fluids was 356.5 minutes (Group A), 402.8 minutes (Group B), and 378.1 minutes (Group C). There was no statistical difference in discharge times from recovery or time to tolerating oral fluids in any of the three groups. PMID- 7485936 TI - The efficacy of adding a continuous intravenous morphine infusion to patient controlled analgesia (PCA) in abdominal surgery. AB - The effect of adding a continuous infusion of morphine 1 mg/hr to patient controlled intravenous analgesia was studied in a randomized double-blind trial. Ninety-six patients scheduled for abdominal surgery were enrolled; 38 received PCA and continuous infusion (PCA + C), 45 received PCA alone and 13 were excluded because of protocol violations. PCA was delivered via an ABBOTT 4200 pump with settings of morphine 1 mg bolus and five-minute lockout in both groups. A separate pump delivered a continuous infusion of morphine 1 mg/ml (PCA + C) or 9% normal saline (PCA) at 1 ml/hr for three postoperative days. Pain was assessed by hourly verbal pain scores (VPS) and daily visual analog pain scores at rest and on movement (VAS(rest), VAS(move)). PCA delivered morphine and the demand to delivery ratio (D/D ratio) were used as indirect indicators of pain. These were assessed during daytime (0800-2200 hours), sleep (2200-0500 hours) and on first waking (0500-0800 hours). Patient demographics were similar. Patients receiving a continuous infusion had lower pain scores during the first 24 hours but not thereafter (VPS P = 0.04, VAS(move) P = 0.02). The PCA group delivered more PCA morphine during 0500-0800 hours and 0800-2200 hours on the first day only. There was no significant difference in the D/D ratio for any time period during the three days. Total morphine delivery was greater in the PCA + C group on the second and third postoperative days (P = 0.009 and P = 0.0001 respectively).(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7485937 TI - Clinical experience with patient-controlled and staff-administered intermittent bolus epidural analgesia in labour. AB - A prospective, randomized study was performed to detail clinical experience with both patient-controlled epidural analgesia (PCEA) and midwife-administered intermittent bolus (IB) epidural analgesia during labour, under the conditions pertaining in a busy obstetric delivery unit. Both methods used 0.125% bupivacaine plus fentanyl, and similar rescue supplementation, although management decisions related to epidural analgesia were made principally by attending midwives. One hundred and ninety-eight women were recruited and data analysed from 167 (PCEA n = 82, IB n = 85). The groups were demographically similar. Median hourly pain scores, ratings of analgesia and satisfaction did not differ. Maximum pain scores were significantly higher in those receiving IB epidural analgesia (P < 0.05). The PCEA group had a significantly higher rate of supplementation and bupivacaine use (P < 0.01), and a longer duration of the second stage of labour (P < 0.03). The relative risk of instrumental delivery with PCEA versus the IB method was 1.57 (CI 1.07-2.38). Experience within our unit with PCEA is contrasted with that of IB epidural analgesia, the method most commonly used; and with that of controlled trials comparing these two methods. PMID- 7485935 TI - A double-blind randomized controlled trial of ginger for the prevention of postoperative nausea and vomiting. AB - The efficacy of ginger for the prevention of postoperative nausea and vomiting was studied in a double-blind, randomized, controlled trial in 108 ASA 1 or 2 patients undergoing gynaecological laparoscopic surgery under general anaesthesia. Patients received oral placebo, ginger BP 0.5g or ginger BP 1.0g, all with oral diazepam premedication, one hour prior to surgery. Patients were assessed at three hours postoperatively. The incidence of nausea and vomiting increased slightly but nonsignificantly with increasing dose of ginger. The incidence of moderate or severe nausea was 22, 33 and 36%, while the incidence of vomiting was 17, 14 and 31% in groups receiving 0, 0.5 and 1.0g ginger, respectively (odds ratio per 0.5g ginger 1.39 for nausea and 1.55 for vomiting). These results were essentially unchanged when adjustment was made for concomitant risk factors. We conclude that ginger BP in doses of 0.5 or 1.0 gram is ineffective in reducing the incidence of postoperative nausea and vomiting. PMID- 7485940 TI - The role of autologous blood transfusion in joint replacement surgery. AB - The efficacy of predeposited autologous blood transfusion (PABT) with and without intra/postoperative blood salvage to reduce or eliminate the need for homologous blood transfusion (HBT) in primary total hip or knee replacement surgery was investigated by retrospective and prospective studies. Depending on the type of surgery, one to three units of PABT eliminated the need for HBT in 50 to 78% of patients, but, intra/postoperative blood salvage alone reduced the need only in 11 to 29%. In contrast, blood salvage, when combined with three units of PABT, eliminated the need for HBT in all patients undergoing primary joint replacement surgery. A cost comparison analysis showed that blood salvage was more expensive than PABT, and therefore it should be limited to patients who had predeposited fewer than three units of autologous blood. PMID- 7485939 TI - Sacral intervertebral approach for epidural anaesthesia in infants and children: application of "drip and tube" method. AB - Sacral intervertebral approach to the epidural space was introduced as an alternative to the caudal approach in infants and children in 1987. We performed single-shot epidural anaesthesia in 200 infants and children with this approach using the "drip and tube" method for identification of the epidural space. Identification of the epidural space was marked by commencement of dripping in the chamber of a micro-drip infusion set which was connected to the epidural needle. With the i.v. extension tube, local anaesthetic solution was injected by the "immobile needle" technique. Overall success rate of the block was 96%. This approach to the epidural space is anatomically easy in infants and children, and application of our "drip and tube" method might make the paediatric single-shot epidural anaesthesia safer and more successful. PMID- 7485938 TI - Effects of midazolam on heart rate variability during surgery under spinal anaesthesia. AB - Effects of midazolam on the cardiac autonomic nervous system were studied by power spectral analysis of electrocardiographic R-R intervals in patients undergoing elective lower abdominal surgery under spinal anaesthesia. Patients were randomly assigned into two groups: 10 patients in group A received spinal anaesthesia only and 10 in group S received spinal anaesthesia and midazolam of 0.05 mg/kg when surgery started. In the frequency domain power spectra, low (Lo; 0.04-0.15 Hz) and high (Hi; 0.15-0.40 Hz) frequency components were integrated to ascertain sympathetic and parasympathetic activity, respectively. There was no intergroup difference in starting time of surgery, ventilatory frequency, arterial pressure, heart rate, mean and variance of R-R interval, and cephalad level of analgesia. In spectral component, Lo decreased (P < 0.05) and Hi/Lo ratio increased (P < 0.05) relative to their baselines in group S. These were also different from group A (P < 0.05). We concluded that intravenous midazolam depressed sympathetic activity to produce a vagotonic cardiac autonomic nervous system under spinal anaesthesia. PMID- 7485942 TI - Fatal cardiovascular collapse following propofol induction in high-risk patients and dilemmas in the selection of a short-acting induction agent. PMID- 7485941 TI - A comparison of the efficacy of dopexamine and dobutamine for increasing oxygen delivery in high-risk surgical patients. AB - Peri-operative increase of oxygen delivery has been shown to reduce mortality in high-risk surgical patients. This study compares the effectiveness of dopexamine and dobutamine when used to increase cardiac output as part of a regimen to increase oxygen delivery. Sixteen surgical patients were randomly allocated to receive either dopexamine or dobutamine, which was increased to a stable dose defined as either oxygen delivery index > 600 ml/min/m2, or tachycardia > 20% above baseline, other dysrhythmias or angina. At this "stable" dose there were significant increases in cardiac index (2.4 +/- 0.2 vs 3.7 +/- 0.3 l/min/m2) and oxygen delivery (380 +/- 73 vs 579 +/- 40 ml/min/m2) in the dopexamine group (P < 0.05); but not the dobutamine group. Five out of eight patients receiving dopexamine and three out of eight receiving dobutamine reached target oxygen delivery. Three dobutamine patients, but no dopexamine patients, had angina or dysrhythmias. In preoperative high-risk surgical patients, dopexamine can allow greater increases in oxygen delivery than dobutamine, due to cardiac effects that limit the dobutamine infusion rate. PMID- 7485943 TI - Accidental intra-arterial adrenaline administration. PMID- 7485944 TI - Blind nasal intubation by monitoring capnography in a neonate with congenital microstomia. PMID- 7485945 TI - Epidural caesarean section in a patient with congenital antithrombin III deficiency. PMID- 7485946 TI - Tension pneumothorax due to intrapulmonary placement of intercostal chest drain. PMID- 7485948 TI - Anaesthesia and moyamoya disease. PMID- 7485949 TI - Prolonged unilateral lower limb paresis following abdominal surgery with epidural and general anaesthesia. PMID- 7485950 TI - Successful treatment of carbimazole-induced agranulocytosis and severe sepsis with granulocyte macrophage colony stimulating factor. PMID- 7485947 TI - Fatal serotonin syndrome following a combined overdose of moclobemide, clomipramine and fluoxetine. PMID- 7485951 TI - Total mechanical alternans in postoperative aortic valve replacement. PMID- 7485952 TI - Mechanical ventilation at moderate altitude. PMID- 7485953 TI - Local anaesthesia for eye surgery. PMID- 7485955 TI - Sprotte needle introducers. PMID- 7485954 TI - Ondansetron and minor oral surgery. PMID- 7485956 TI - Propofol and epilepsy. PMID- 7485958 TI - A problem with a Sechrist air/oxygen blender device. PMID- 7485959 TI - Anaesthesia in epiglottitis. PMID- 7485957 TI - The CO2 laser and latex armoured tube. PMID- 7485960 TI - A dangerous problem with plastic Yankauer suckers. PMID- 7485961 TI - Convulsion following spinal anaesthesia. PMID- 7485962 TI - Anaphylactoid reaction to Haemaccel. PMID- 7485963 TI - Comparison of measurement techniques: agreement is not correlation. PMID- 7485964 TI - A new measure of anaesthetic potency. PMID- 7485965 TI - Use of an infant feeding tube in fibreoptic intubation. PMID- 7485966 TI - Clinical indicators: uses and misuses. PMID- 7485967 TI - Enzymatic-chemical preparation of quinoxaline derivatives from L-amino acids for gas chromatographic-mass spectrometric analyses. AB - We report on an enzymatic-chemical method for the specific preparation of L-amino acid-derived quinoxalinols suitable for the sensitive estimation of 13C- or 2H label enrichment by gas chromatography-mass spectrometry: Amino acid fractions are isolated from physiological fluids by ion-exchange chromatography. The compound of interest is converted to the corresponding 2-oxo acid by treatment with an L-amino acid dehydrogenase of the desired specificity. Reaction of the 2 oxo acid with o-phenylenediamine yields the quinoxaline derivative. Isotopic label enrichment is then determined by gas chromatographic-mass spectrometric analysis of the O-trimethylsilyl derivative using an ammonia-chemical ionization mode and selected ion monitoring of the quasi-molecular ions [MH]+ and [MH + n]+ (n = number of labeled positions). Details for application of various generally available L-amino acid dehydrogenases (L-ala DH, L-glu DH, L-leu DH, L-phe DH) are presented. The method was used, e.g., for serum analysis in in vivo studies on the decarboxylation rates of branched-chain L-[1-(13)C]-amino acids. PMID- 7485968 TI - Quantitation of digoxigenin-labeled DNA hybridized to DNA and RNA slot blots. AB - Quantitation of message from low-abundance mRNAs and limited availability of tissues requires sensitive methods for probe detection, accurate methods for quantitation of signal, and the ability to strip and reprobe membranes. A random primed, digoxigenin-labeled probe from cDNA of FMO1, an isoform of the flavin containing monooxygenase gene family, from rabbit was used in the evaluation and optimization of the Genius system for quantitation of signal from DNA and RNA slot blots. Criteria for optimization were a low signal to noise ratio, a linear increase in density of signal vs nuclei acid concentration of bands on X-ray film, complete stripping of membranes, and reproduction of the initial banding pattern upon rehybridization. A low signal-to-noise ratio was obtained with an aqueous prehybridization/hybridization solution. DNA slot blots were successfully quantitated before and after alkaline stripping from positively charged membranes. RNA slot blots were subject to excessive and uneven loss of RNA from the membranes during stripping procedures. Reliable quantitation for more than one cycle of detection required highly charged nylon membranes, and careful tailoring of RNA fixation methods and alkaline stripping conditions. PMID- 7485969 TI - Epitope tagging of the human endoplasmic reticulum HSP70 protein, BiP, to facilitate analysis of BiP--substrate interactions. AB - We modified BiP, the resident endoplasmic reticulum (ER) heat shock protein 70, to contain an epitopetag sequence close to the C-terminus (BiP-tag); the epitope is derived from an influenza hemagglutinin (HA) subtype and is recognized by the monoclonal antibody 12CA5. This antibody both immunoprecipitates BiP-tag and detects it on Western blots. Using transient expression of cDNAs in COS cells, we studied the interaction of BiP-tag with several membrane proteins. Consistent with previous work on BiP, BiP-tag bound poorly and transiently to newly made wild-type influenza HA glycoprotein and strongly and irreversibly to an HA mutant that misfolds and is retained in the ER. Most newly made erythropoietin receptor (EPO-R) polypeptides are retained in the ER and degraded there; we show here that, in cotransfected COS cells, newly made EPO-R is bound to BiP-tag prior to its degradation. Thus, by several criteria the BiP-tag molecule is fully functional in binding newly made proteins. Because it can be immunoprecipitated by a readily available antibody, it offers several advantages to the study of protein folding in the ER and the role of chaperones in this process. PMID- 7485970 TI - Incorporation of 5-iodo-2'-deoxyuridine and 5-bromo-2'-deoxyuridine into rodent DNA as determined by neutron activation analysis. AB - Using 5-iodo-2'-deoxyuridine (IdU) and 5-bromo-2'-deoxyuridine (BrdU) as DNA precursors, neutron activation analysis (NAA) of iodine and Br was developed as a quantitative method for determining DNA synthesis. Endogenous rodent tissue concentrations of bromine (Br) and iodine ranged 100-fold from a low of 0.06 microgram of iodine/g of rat gastrointestinal tract (GIT) to a high of 5.99 micrograms of Br/g of rat kidney. All 10 rodent tissues had concentrations of Br 4 to 76 times higher than those of iodine. Rat hepatic Br concentrations could be reduced 17-fold by dietary and pharmacological methods. Female Fischer 344 rats and male C57BL/6 mice were given 4-8 intraperitoneal injections of either IdU or BrdU as a DNA precursor. Tissue clearance of iodine in IdU-treated rodents was both faster and more complete (in mice 4 and in rats 17 h or less) than Br clearance from BrdU-treated rodents (at 162 h nonincorporated Br label still remains). In rat liver, lung, and kidney, the iodine label incorporated from IdU into DNA was stable for at least 162 h. The incorporation ratio is defined as the microgram halogen/tissue for either IdU- or BrdU-treated rodents divided by the microgram halogen/g tissue of untreated rodents. NAA-based studies of DNA synthesis gave high incorporation ratios in rat liver (5.3), rat lung (6.7), rat GIT (19.0), rat spleen (24.0), mouse GIT (10.1), and mouse spleen (25.8). PMID- 7485971 TI - Preparative and analytical capillary zone electrophoresis analysis of native endorphins and enkephalins extracted from the bovine pituitary: mass spectrometric confirmation of the molecular mass of leucine enkephalin. AB - One proopiomelanocortin peptide [beta-endorphin (BE)] and two proenkephalin A peptides [methionine enkephalin (ME) and leucine enkephalin (LE); LE derives also from proenkephalin B] were searched for in a bovine pituitary extract by capillary zone electrophoresis (CZE) and liquid secondary-ion mass spectrometry (LSIMS). A bovine pituitary homogenate was subjected to acid precipitation/centrifugation and solid-phase extraction of peptides using an octadecyl-silyl disposable cartridge. The peptide-enriched fraction was subjected to CZE at pH 2.5 and at pH 5.5., and fractions were collected under preparative CZE conditions within defined time windows where synthetic BE, ME, and LE migrate. The resolving power of CZE was demonstrated by collecting biological fractions at pH 5.5 under preparative conditions and by subsequently analyzing these fractions at pH 2.5 under analytical conditions. Preparative CZE was further performed at pH 2.5 for fractions collected at pH 5.5. LSIMS analysis of this second-dimensional CZE fraction revealed the appropriate protonated molecule ion [(M + H)+, m/z 556.4] of LE. PMID- 7485972 TI - Inverted terminal repeats permit the average length of amplified DNA fragments to be regulated during preparation of cDNA libraries by polymerase chain reaction. AB - A simple polymerase chain reaction (PCR)-based technique for construction of cDNA libraries starting with very small amounts of cells or tissues is described. The technique is based on the insertion of inverted terminal repeats into amplified cDNAs which permit short molecules to generate "pan"-type structures at each cycle of PCR amplification and thus to escape annealing with primers. This allows one to avoid amplification of primer dimers and makes it possible to perform oligonucleotide tailing of the first cDNA strands followed by PCR amplification in the same tube. Moreover, the average size of amplified cDNAs can be controlled by varying the primer concentration. PMID- 7485973 TI - Reverse isoelectric focusing procedure resolves charge variants of basic proteins. AB - Several powerful methods exist which effectively resolve proteins based on their differences in overall change. Among these, two-dimensional gel electrophoresis, a method combining isoelectric focusing with sodium dodecyl sulfate polyacrylamide gel electrophoresis, can resolve over 1000 proteins with isoelectric points in the range of pH 4-7. However, resolution of basic proteins is poor, even when the pH gradient is extended to a higher pH. Nonequilibrium pH gradient electrophoresis was developed as an alternative procedure to resolve basic proteins. However, samples must be dissolved in urea prior to analysis and this method requires 2000-8000 V-h for optimal resolution. In this report we demonstrate that a simple, rapid procedure, based on nonequilibrium pH gradient electrophoresis, can resolve the single charge variants of two fungal enzymes with isoelectric points greater than 8. This procedure utilizes ready-made materials and requires less than 500 V-h to complete (total run time is less than 2 h). PMID- 7485975 TI - Detection of insulin aggregates with pulsed-field gradient nuclear magnetic resonance spectroscopy. AB - Self-association of metal-free human insulin in aqueous solution at pH 9.4 has been studied using pulsed-field gradient (PFG) NMR spectroscopy. The diffusion coefficients were measured for two different insulin concentrations, 0.72 mM in which the dimer is the predominant species and 3 mM which is a mixture of dimer and tetramer. In the more concentrated solution, exchange between the two aggregates is slow on the NMR chemical shift time scale. As a result, the dimer and tetramer give rise to separated resonances in the aromatic region of the 1H NMR spectrum. The diffusion coefficient determined for the tetramer by PFG NMR is 1.07 x 10(-6) cm2 s-1 at 298 K, while that for the dimer is 1.38 x 10(-6) cm2 s 1. The hydrodynamic diameters calculated for the tetramer and dimer are 46 and 36 A, respectively, which are in good agreement with those measured by dynamic light scattering. The advantage of PFG NMR for the measurement of insulin aggregation in these solutions results from the chemical shift selectivity which allows diffusion coefficients to be directly calculated for the different aggregates. The T1 and T2 relaxation times of the dimer and tetramer aromatic protons were also measured in order to optimize the parameters of the PFG NMR experiment and correctly interpret the results of the measured spectra. PMID- 7485974 TI - Homogeneous quantitative assay of hepatitis C virus RNA by polymerase chain reaction in the presence of a fluorescent intercalater. AB - We have developed a homogeneous quantitative assay of DNA/RNA by performing PCR in the presence of an oxazole yellow derivative, a fluorescent DNA intercalative dye, and monitoring the fluorescence intensity of the PCR reaction mixture during PCR cycles. We have demonstrated the applicability of this assay by use it to quantify hepatitis C virus (HCV) RNA of serum samples from patients with chronic hepatitis C. This assay gave efficient and reproducible results in a clinically useful dynamic range below 10(-6) copies of HCV RNA for interferon therapy. PMID- 7485976 TI - Analysis of monoclonal antibody chimeric BR96-doxorubicin immunoconjugate by sodium dodecyl sulfate-capillary gel electrophoresis with ultraviolet and laser induced fluorescence detection. AB - Sodium dodecyl sulfate-capillary gel electrophoresis (SDS-CAGE), the capillary version of SDS-polyacrylamide-based slab gel electrophoresis, has been utilized for the separation and analysis of monoclonal antibody chimeric BR96 and the corresponding immunoconjugate prepared between BR96 and the anticancer drug doxorubicin (BR96-DOX). SDS-CAGE was performed in a coated capillary column filled with a polymer solution-based gel network matrix. Two detection formats, a uv absorbance detector and an argon-ion laser-based fluorescence detector, were incorporated into this system, providing complementary information for the determination of conjugated species. Both monoclonal antibody and immunoconjugates were studied in their native, denatured, and denatured and reduced states, respectively. Six peaks were identified following separation of the denatured BR96-DOX. These peaks were confirmed to correspond to all the possible conjugated species as expected. Analysis of the resulting "fingerprint" maps indicated that the light, heavy, and light-heavy chain conjugates are the predominant species. SDS-CAGE offers an alternative approach to the conventional slab gel electrophoresis and other chromatographic techniques, providing rapid, efficient, sensitive, and accurate information for the analysis of antibody and bioconjugates. PMID- 7485977 TI - A fluorometric assay for quantitating DNA strand breaks during apoptosis. AB - Few techniques exist for quantitating DNA fragmentation during apoptosis. Our aim was to develop a quantitative assay for DNA fragmentation in apoptosis by enzymatically labeling DNA with a fluorescent dideoxynucleotide. Terminal deoxynucleotidyl transferase was used to enzymatically label 3'-OH DNA ends with fluorescein-12-dideoxyuridine triphosphate in an assay referred to as fluorophore end-labeling. Because only one labeled dideoxynucleotide can be added per 3'-OH end of DNA, the fluorescence intensity is directly proportional to the number of DNA strand breaks. The sensitivity and validation of this approach were first established in isolated calf thymus DNA treated with the endonuclease, DNase I; and excellent correlation was observed between fluorophore end-labeling and an isotopic approach to quantitate 3'-OH ends of DNA. Quantitation of DNA strand breaks was then obtained in nuclei isolated from hepatocytes undergoing apoptosis using fluorescent digitized microscopy, flow cytometry, and fluorometry. In addition to its quantitative aspects, fluorophore end-labeling proved to be quite sensitive as it detected DNA strand breaks prior to the morphologic changes of apoptosis or the development of the hypodiploid state as assessed by fluorescence microscopy and flow cytometry, respectively. This assay should prove useful for studying the molecular mechanisms leading to DNA cleavage during apoptosis. PMID- 7485978 TI - Quantitative polymerase chain reaction with enzyme-linked immunosorbent assay detection of selectively digested amplified sample and control DNA. AB - A quantitative polymerase chain reaction (PCR) method for the exact quantitation of DNA is described that does not require radioactive labeling or electrophoretic separation of product species, thereby avoiding hazardous and time-consuming procedures which have so far impeded routine use of PCR, in particular in the clinical laboratory. Sample and internal control DNA are competitively amplified in a one-tube nested PCR. The control DNA differs from the sample DNA by only two base pairs which change a single restriction site for a new one. Thereby, it is possible to discriminate the two PCR product species by selective restriction enzyme digestion (RED). Because inner PCR primers were labeled with biotin and digoxigenin, respectively, nested PCR products can be immobilized on avidin coated microtiter plates and quantitated separately by enzyme-linked immunosorbent assay (ELISA) techniques. We demonstrate here that with the combination of selective restriction enzyme digestion and ELISA (RED-ELISA) sample DNA ranging from nanomolar to attomolar concentrations can be quantitated within +/- 10%. This procedure can be easily adapted for quantitation of other PCR products. It is suitable for rapid and automatic screening of many samples in parallel, e.g., for detection and quantitation of pathogens, for quantitation of gene copy numbers or for gene expression after reverse transcription. PMID- 7485979 TI - Spectrophotometric determination of 6-aminopenicillanic and 7 aminocephalosporanic acids as the Schiff bases with para dimethylaminobenzaldehyde in the presence of sodium dodecyl sulfate micelles. AB - Yields of colored Schiff base products of the reversible condensation of 6 aminopenicillanic and 7-aminocephalosporanic acids with para dimethylaminobenzaldehyde in a weakly acid aqueous solution strongly increase in the presence of micelles of sodium dodecyl sulfate. The binding constants of reactants and products of both reactions to micelles were calculated from the concentration profiles of micellar effects on rates and equilibrium of the condensation reactions. The observed micellar-induced shifts of equilibrium are accompanied by strong acceleration of the forward and small retardation of backward processes. Some deviations from the theoretical profiles due to saturation of micelles with reactants were observed. The colored form of the reaction product was found to be the protonated Schiff base. Its stability in time is strongly enhanced in the micellar medium. The spectrophotometric determination of title compounds under optimum micellar conditions possesses ca. 300 times higher sensitivity than the standard procedure. PMID- 7485981 TI - A procedure for the determination of monothiols in the presence of dithiothreitol -an improved assay for the reduction of disulfides. AB - 4,4'-Dipyridyl disulfide (4-PDS) and sodium arsenite have been used to develop a rapid and sensitive assay for monothiols in the presence of dithiothreitol (DTT). The procedure is similar to that of Zahler and Cleland (J. Biol. Chem. 243, 716 719, 1968) but involves the use of a lower pH and 4-PDS, which is more effective than 5,5'-dithiobis-(2-nitrobenzoic acid) at low pH. Background reactions that interfere with the determination of slow reacting thiol groups are much reduced in the new assay procedure. The reduction of oxidized glutathione by DTT at three different pH values and a partial and the complete reduction of bovine serum albumin by DTT have been characterized in order to demonstrate the use of this procedure. In the first two cases monothiol groups were readily detected without interference from DTT. Lower than expected numbers of thiol groups in the case of three reduced and denatured model proteins appeared to reflect their partial complexation by arsenite. Equilibrium and kinetic constants for the formation and dissociation of the DTT-arsenite complex from pH 5 to 8 are presented. PMID- 7485980 TI - Continuous measurement of phospholipase A2 activity using the fluorescent probe ADIFAB. AB - A new method is described for the continuous quantitation of phospholipase A2 (PLA2) activity with greatly improved sensitivity compared to existing techniques. The method utilizes a fluorescent probe to detect the release of fatty acid monomers (free fatty acids) into the aqueous phase. The fluorescent probe ADIFAB, which is the acrylodan derivative of rat intestinal fatty acid binding protein, exhibits a change in the ratio of its fluorescence upon binding medium- to long-chain native fatty acids. ADIFAB was used to measure the hydrolysis of artificial and natural membranes using PLA2s from porcine pancreas, Naja mocambique mocambique, Crotalus durissus terrificus, and bee venom. Total phospholipid hydrolysis was determined from the free fatty acid concentration using membrane/water partition coefficients, also measured using ADIFAB. The results indicate that continuous monitoring of natural substrates can be determined with a sensitivity limit of less than 1 pmol/min, a more than 10(4) fold increase in sensitivity over the most commonly used pH stat method. PMID- 7485982 TI - Development of a melanin-based high-performance liquid chromatography stationary phase and its use in the study of drug-melanin binding interactions. AB - Drug-melanin interactions were studied using a melanin-based HPLC column. Two approaches were chosen for the preparation of the stationary phases: covalent coupling of synthetic L-dopa melanin and in situ polymerization of L-dopa. Retention of a series of phenothiazines on melanin-based stationary phases was attributed to binding to melanin. Frontal affinity chromatography experiments on one melanin-based column allowed us to calculate the affinity and binding capacity of chlorpromazine and promethazine. A competition was observed between chlorpromazine and haloperidol which was qualitatively consistent with previously described results. Data indicated that the interaction was not a simple competition at one site. PMID- 7485984 TI - Codon-based combinatorial alanine scanning site-directed mutagenesis: design, implementation, and polymerase chain reaction screening. AB - Combinatorial alanine scanning mutagenesis is a powerful tool for the exploration of protein structure-function relationships. Unfortunately, combinatorial alanine replacement of multiple residues using standard site-directed mutagenesis is restricted to a subset of amino acids. To circumvent this limitation, an efficient procedure for combinatorial site-specific replacement by alanine of any residue in a given protein sequence has been established. The method, which involves simple procedures and commonly used materials, is based upon the use of codon-based mutagenesis. A defined ratio of alanine to wild-type codon was introduced at each predetermined triplet using the "column-splitting" technique during oligonucleotide synthesis. High-throughout genetic screening of mutant libraries was facilitated by the incorporation of diagnostic restriction sites at targeted codons followed by a PCR-based screening procedure. The method was tested on a set of 13 residues located at the interface between the variable domains of a Fab fragment of an antibody. The occurrence of alanine substitution was found to be comparable to the statistically predicted distribution. PMID- 7485983 TI - Measurement of protein in natural rubber latex. AB - Latex from the Brazilian rubber tree (Hevea brasiliensis) is the source of virtually all commercial natural rubber (cis-1,4-polyisoprene). Products made from natural rubber latex include gloves, condoms, and hundreds of different medical devices. However, recent reports of widespread life-threatening latex associated Type I allergies have focused attention on latex proteins as serious allergens. In this paper, we describe a method that permits accurate and reproducible determination of protein in latex and that includes a procedure for solubilizing latex proteins and removing rubber. Also, we show that interfering substances in latex can be removed by precipitating proteins with sodium deoxycholate and trichloroacetic acid, and that latex proteins can be recovered and quantified with 2,2'-bicinchoninic acid. PMID- 7485985 TI - Immobilized liposome chromatography for analysis of interactions between lipid bilayers and peptides. AB - Liposomes were sterically immobilized in gel beads to be used for isocratic chromatographic analysis of interactions between lipid bilayers, amino acids, and water-soluble peptides. Tryptophan was more retarded on the immobilized liposomes than were other amino acids, and peptides with C-terminal cysteine were much more retarded than were peptides with serine or aminobutyric acid instead of the cysteine. Peptide sequences could affect the peptide-liposome interactions. For peptides corresponding to polypeptide segments of a membrane protein, the human red cell glucose transporter (Glut1), the retention volumes increased with decreasing water-to-oil transfer free energy of the peptides and were related to the transfer free energy distribution within the peptides as shown by hydropathy plots and to the presence of cysteine. Among these peptides, the partially hydrophobic peptides 125GRFIIGVYCG134 and 201CIVLPFCPES210 and the hydrophilic peptide 421CFQYVEQLC429 were most strongly retarded on immobilized phosphatidylcholine liposomes. The results indicated that the peptide-liposome interactions were mainly of hydrophobic nature and that the structure of the interfacial head-group region of the lipid bilayers was important. PMID- 7485986 TI - Immunoaffinity chromatography on antibodies immobilized on nitrocellulose powder. AB - A method is described for the use of nitrocellulose powder as a solid phase in a chromatographic procedure, for the immunoaffinity isolation of proteins. Two different immunoglobulins (Igs), anti-Datura innoxia lectin and anti-tyrosinase, were coupled to particulate nitrocellulose. A single step was then needed for purification to homogeneity of both D. innoxia lectin and mouse tyrosinase. Chaotropic and acidic agents proved to be effective in eluting antigens from Ig nitrocellulose columns. The binding capacity of particulate nitrocellulose was around 3 mg Ig per milliliter of nitrocellulose, while the purification yields of the two proteins investigated under various eluting conditions were higher than 75%. The applicability of this method in the identification of metabolically labeled proteins in crude extracts is also demonstrated. Purification of proteins by affinity chromatography on their specific Igs linked to nitrocellulose matrices could be performed in both batch and column. The major advantages of this new method for purification of proteins are its rapidity, the reusability of the affinity matrices, and the high yields of purified protein obtained. The method could be seen as an alternative to the widely used immunoprecipitation technique. PMID- 7485987 TI - Electrospray ionization-mass spectrometry as a tool for characterization of glutathione S-transferase isozymes. AB - Reversed-phase high-performance liquid chromatography coupled to electrospray ionization-mass spectrometry (ESI-MS) was used for the first time to determine the molecular masses of nine rat liver cytosolic glutathione S-transferase (GST) subunits. The precision of the measurements was +/- 3-4 mass units which, in practice, allowed discrimination between monomers differing by more than 8 Da. Mass accuracy was improved by replicates in the measurements. Comparison of experimental values to cDNA or protein-deduced data available reveals slight differences. N-terminal sequence analyses and interstrain mass comparisons tend to show that primary structures of liver cytosolic GSTs are well conserved from Sprague-Dawley to Wistar rats. Moreover, ESI-MS analysis enabled identification of two minor additional subunits present in both strains, one of which belongs to the mu-class. In addition to rapid and accurate mass determination of GST monomers, and direct determinations achieved on heterodimeric forms, this technique provides precise information on minor structural differences or modifications of these proteins. As such, it constitutes a useful tool for rapid characterization of purified GSTs in comparative studies. PMID- 7485988 TI - Chromogenic assay for phospholipase C from Bacillus cereus. AB - A standard chromogenic assay for phosphatidylcholine determination has been modified in the development of a facile method for measuring the rate parameters (Km and Vmax) for phosphatidylcholine-preferring phospholipase C from Bacillus cereus (PLCBc). The assay is applied to the determination of the Ki for a known inhibitor of PLCBc. PMID- 7485989 TI - Preparation of 13C-labeled polyunsaturated fatty acids by an arachidonic acid producing fungus Mortierella alpina 1S-4. AB - Three types of 13C-labeled polyunsaturated fatty acids (PUFAs) were produced in mycelia of an arachidonic acid-producing fungus, Mortierella alpina 1S-4, when grown with 13C-labeled glucose or fatty acid methyl esters. We analyzed them by negative-ion fastatom bombardment mass spectrometry. (i) PUFA in mycelia grown with [1-(13)C]glucose or [2-(13)C]glucose was a mixture of molecules containing zero to six labeled carbons. The content of 13C in the total carbon atoms of PUFA, such as arachidonic (20:4 omega 6), dihomo-gamma-linolenic (20:3 omega 6), gamma-linolenic (18:3 omega 6), and linoleic (18:2 omega 6) acids, in the mycelia grown with [1-(13)C]-glucose or [2-(13)C]glucose was about 11 or 15%, respectively. More than 80% of these PUFA had more than one 13C atom. (ii) When the fungus was grown with [U-13C]glucose, the content of PUFA molecules, all carbons of which were 13C-labeled, was about 20%. The percentage of 13C in PUFA from [U-13C]glucose was 78-83%, over 98% of which had more than one 13C atom. The 13C atoms in the labeled fatty acids formed from 13C-labeled glucose were thought to be distributed at random by a two carbon-unit. (iii) The 13C atom of PUFA in mycelia grown with 1-(13)C-labeled fatty acid methyl esters was highly localized. More than half of the 20:4 omega 6 molecules had only one 13C atom at C-3, C-5, or C-7 when the fungus was grown with methyl [1-13c]stearate, methyl [1 (13)C]palmitate, or methyl [1-(13)C]-myristate, respectively. PMID- 7485990 TI - A slot blot method for detection of ultraviolet photoproducts in DNA. AB - A simple new method was established to detect the major uv photoproducts in DNA. A slot blot method, involving the use of T4 DNA polymerase-associated (3'--> 5') exonuclease digestion of uv-irradiated DNA, was used to detect (primarily) cyclobutane pyrimidine dimers and pyrimidine-pyrimidone (6-4) dimers. Hydrolysis of DNA by this enzyme is quantitatively blocked by both of these photoproducts. To detect (primarily) (6-4) dimers, Escherichia coli DNA photolyase was utilized to specifically reverse cyclobutane pyrimidine dimers. PMID- 7485991 TI - Determination of ascorbic acid and dehydroascorbic acid in plasma by high performance liquid chromatography with coulometric detection--are they reliable biomarkers of oxidative stress? AB - The concentrations of the hydrophilic antioxidants ascorbic acid and dehydroascorbic acid in plasma for some time have been considered possible biomarkers of oxidative stress. However, several problems are associated with the accurate measurement of these two compounds. We have developed and validated a selective and reproducible high-performance liquid chromatographic method for the quantification of ascorbic acid and dehydroascorbic acid in plasma. The method meets the requirements of a reliable routine analysis. The plasma samples are stabilized with 5 mM metaphosphoric acid, centrifugated at 4 degrees C before HPLC analysis. For ascorbic acid analysis, the sample pH is adjusted to 2.6, whereas for total ascorbic acid measurement dehydroascorbic acid is reduced to ascorbic acid using dithiothreitol for 5 min at pH 6.2 after which the sample pH is adjusted to 2.6. The samples are analyzed on a reversed-phase system using coulometric detection. Dehydroascorbic acid concentrations ae calculated by subtraction. Within- and between-day coefficients of variation for the complete assay were in the range of 4-8 and 3-6% for ascorbic acid and total ascorbic acid, respectively. The stability of ascorbic acid was monitored under various conditions including storage and the implications as well as the reliability of ascorbic acid as a biomarker are discussed. PMID- 7485992 TI - Quantitative determination of 8-hydroxyguanine and guanine by isotope dilution mass spectrometry. AB - 8-[4,5,6,8-13C4]Hydroxyguanine and [4,5,6-13C3]guanine were prepared by total synthesis. The labeled compounds were used as internal standards in a gas-liquid chromatographic-mass spectrometric method for quantitative determination of 8 hydroxyguanine and guanine in DNA. The approach used made it possible to directly determine the ratio between 8-hydroxyguanine and guanine, expressed as the number of molecules of 8-hydroxyguanine per 10(5) molecules of guanine, in the sample of DNA analyzed. A method for trimethylsilylation of 8-hydroxyguanine and guanine at room temperature was developed. Use of this method for derivatization gave levels of 8-hydroxyguanine in calf thymus DNA which were lower than those recently found by gas chromatography-mass spectrometry but similar to those determined by high performance liquid chromatography with electrochemical detection. PMID- 7485993 TI - Fast and specific labeling of antibody fragments with multiple atoms of heavy metal radioisotopes. PMID- 7485994 TI - Stable-isotope dilution studies of an intact protein using high-performance liquid chromatography/chemical reaction interface mass spectrometry. PMID- 7485995 TI - A method for purification of DNA species by high-speed centrifugation of agarose gel slices. PMID- 7485996 TI - Enhanced resolution of viral double-stranded RNA segments by electrophoresis at low temperatures. PMID- 7485997 TI - Expression of the chicken retinoid X receptor-gamma gene in migrating cranial neural crest cells. AB - We have used in situ hybridisation to whole chick embryos with digoxygenin labelled probes to investigate the distribution of RXR-gamma transcripts during neural crest cell migration in the developing head and the anterior of the trunk (the vagal region), where neural crest cells make a substantial contribution. We have found that RXR-gamma transcripts are a good marker for migrating neural crest cells in the chick embryo. RXR-gamma transcripts were first detected in cells that had recently emerged from the neural crest, providing an earlier marker for neural crest cells than the HNK-1 epitope. The pattern of RXR-gamma transcript distribution is dynamic in the developing chick head, and changes in a pattern which is coincident with the migration of cells containing RXR-gamma transcripts and the gradual restriction of RXR-gamma transcripts to specific differentiating neural crest derivatives. Transcripts appeared to be present initially in migrating neural crest cells thoughout the developing head, but gradually became restricted to some crest-derived populations and absent from others. By stage 15, RXR-gamma transcripts were not detectable in neural-crest derived ectomesenchymal cells, although they were still found in cells contributing to the cranial ganglia and their roots. PMID- 7485999 TI - Number and size spectra of non-myelinated axons of human premolars. AB - The aim of this study was to determine the number and size of apical non myelinated (C) axons of healthy human premolars. The material was derived from a large collection of specimens prepared for a previous quantitative investigation on the myelinated (A) axons of human premolars. A total of 16 teeth (six maxillary first and five each of mandibular first and second premolars), removed from adolescents for orthodontic reasons, were used. Root discs of about 0.6 mm thickness were prepared at about 2 mm cervical to the root apex and processed for light and electron microscopy. The number of non-myelinated axons was determined by taking a total census of such fibres that could be identified and reconstructed by standardized composite electron micrographs from each root disc. The measurement of axons was done on a statistically representative sample of axons (n = 1810) using an electronic image processing unit. The 16 teeth had an average of 2000 +/- 1023 non-myelinated axons at the juxta-apical level (range 534-3912). The average diameter of the non-myelinated axons was found to be 0.5 +/- 0.4 microns (range 0.05-2.4 microns). PMID- 7485998 TI - The effects of retinoid status on TGF beta expression during mouse embryogenesis. AB - In a previous study we investigated the effects of RA excess on TGF beta protein localization in early postimplantation stages of mouse development. Here we extend this investigation by comparing the effects of retinoid deficiency with those of excess, and by comparing the effects of altered retinoid status on TGF beta protein and RNA transcript distribution. In vitamin A-deficient embryos, TGF beta 1 RNA and protein distribution were both unaltered compared with controls; conversely, TGF beta 2 protein levels were reduced while RNA levels remained normal. In RA-treated embryos, the previous study showed that intracellular TGF beta 1 levels were decreased, while those of extracellular TFG beta 1 were initially decreased but subsequently increased; here we found that TGF beta 1 RNA transcript levels were reduced following exposure to RA excess. TGF beta 2 showed a clear disparity between the effects of RA excess on protein and RNA transcript levels: RNA transcript distribution was unchanged or showed a slight increase in RA-treated embryos, whereas the previous results showed greatly reduced protein levels. The new results provide further evidence for interaction between retinoids and TGF beta s during mouse development, and indicate that retinoids are capable of differentially regulating TGF beta isoforms through mechanisms involving different stages in the process of TGF beta synthesis and secretion. The long-term nature of the effects of transient exposure to RA excess suggests that the mechanisms of RA-TGF beta interaction may be indirect. PMID- 7486000 TI - Organogenesis of the orbital glands in the lizard Podarcis s. sicula: a histological, histochemical and ultrastructural study. AB - The orbital glands of the lizard Podarcis s. sicula are represented by the anterior and posterior lacrimal glands and the Harderian gland. The anlage of the Harderian gland appears on about the 22nd day of development in the form of a short tubule projecting from the conjunctival epithelium. This event is coincident with the appearance of the nictitating membrane. At this stage the mesenchymal cells surrounding the glandular blastema proliferate at a high rate and form a definite sac, later occupied by both the Harderian gland and the anterior lacrimal glands. At the 26th day of development, the glandular blastema forms acini at its distal end. The prospective glandular cells are not yet differentiated histologically. At the 36th day of development, differentiated serous glandular cells become visible. At the 41st day of development, the acini fill up the preformed mesenchymal sac. Only at this stage does the most medial part of the gland differentiate into mucous-secreting anterior lacrimal gland. At the same time, a small primordium of the posterior lacrimal gland can be seen in the posterior commissure of the eye. The appearance of junctional complexes between epithelial cells and mesenchymal cells in the early developmental stages supports the role of the mesenchyme in the differentiation of the glandular cells. Since the glandular anlage differentiates laterally into Harderian gland and medially into anterior lacrimal gland, spatial and temporal differences seem to exist in the inductive process. Furthermore, a concentration gradient of the inductive substance(s) may be envisaged, since an intermediate zone is present between the Harderian gland and the anterior lacrimal gland, consisting of mixed glandular cells containing both mucous and serous secretory granules. PMID- 7486001 TI - The development of fibrocartilage in the rat intervertebral disc. AB - The development of fibrocartilage in rat lumbar intervertebral discs has been correlated with an immunohistochemical analysis of the changing distribution of extracellular matrix components. Disc anlagen were first recognised by embryonic day 14 as segmental cell condensations. By E16, the notochord formed a series of bulges, each representing a future nucleus pulposus, and the annulus fibrosus had differentiated in the disc anlagen. The inner part of the annulus was composed of cartilage which linked that of adjacent vertebral bodies. The outer part was fibroblastic, with layers of parallel fibroblasts. The long axes of the cells in successive layers lay at an angle of approximately 90 degrees to each other. This criss-cross orientation of cells preceded the oriented deposition of collagen fibres to form the lamellae. Disc anlagen were immunolabelled weakly for types I and III collagen, chondroitin 6-sulphate and dermatan sulphate. Later tissue differentiation was marked by the appearance of type II collagen, chondroitin 4 sulphate and keratan sulphate in the inner annulus. These components also appeared in the outer annulus, but only in adult animals, and indicated metaplastic change in the lamellar fibroblasts. Fibrocartilage in the nucleus pulposus was only seen in old animals, and the origin of the tissue was less clear. However, the fibrocartilage cells appeared to be derived from the cartilage end plate and/or from the inner annulus. We conclude that fibrocartilage in the intervertebral disc is derived from several sources and that the radial distribution patterns of extracellular matrix components in the adult disc are explained by the embryonic origins of its parts. PMID- 7486002 TI - Afferents to different layers of the dorsolateral isocortex in rats. AB - Fluorescent somatopetal tracers were used to infiltrate, by diffusion rather than injections, the dorsolateral cortex of one hemisphere in rats. In different animals the tracers penetrated into the cortex to different depths. We found several interesting features of the commissural system: first, there were no areas without commissural neurons. At least a few labelled cell bodies were present in a single-cell layer also in "acallosal" cortical areas. Secondly, there is a considerable variety of laminar distribution patterns of labelled perikarya in different areas. Thirdly, some cortical fields, which cytoarchitecturally appear uniform, can be subdivided according to different distributions of cell bodies with commissural projections. Fourthly, when only supragranular layers were infiltrated, labelled cell bodies were present mainly in the supragranular layers of the contralateral cortex. Infiltration of the first layer alone did not label any neurons in the contralateral cortex but did label neurons in layer VIb ipsilaterally. In the subcortex, the labelled perikarya were found in the structures already known to project directly to the cortex. In rats with the tracer restricted mainly to the supragranular layers, a conspicuously reduced labelling was found in the basal forebrain and the thalamus. In the thalami of those animals, labelled neurons were found only in paralamellar nuclei. The high sensitivity of the tracer used, together with infiltration of the entire dorsolateral cortex, allows us to conclude that probably all sources of innervation of the isocortex in rats have been seen. PMID- 7486003 TI - Cell death in the olfactory epithelium. AB - In the nervous system of vertebrates the olfactory epithelium presents unique cytological characteristics. In the olfactory mucosa, olfactory neurons die and are replaced from undifferentiated neuroblasts over the entire life span of the animal. It remains unclear whether these neurons die as a result of a direct insult from the environment or in fulfillment of a physiological program of cell death. We have studied the distribution and the characteristics of cell death in the olfactory epithelium of normal, adult rats. The olfactory epithelium contains pycnotic bodies resembling those described for thymocytes undergoing terminal apoptotic changes. These appear at all levels in the epithelium, under both light and electron microscopes and can also be demonstrated after vital staining with acridine orange. Chromatin condensation into large blocks, often located at the nuclear periphery, is a morphological hallmark of the nuclei of mature olfactory neurons, which also present an increase in electron density of the cytoplasm. After non-radioactive in situ labeling of fragmented DNA, the nuclei of olfactory neurons are positive. Under the same reaction conditions (mild protease digestion), most of the nuclei of the supporting and basal cells are negative. In vivo incorporation of 5-bromouridine, a marker of RNA synthesis, is also lower in olfactory neurons than in basal and supporting cells. These findings suggest that olfactory neurons are committed very early to physiological cell death. PMID- 7486004 TI - Influence of maternal adrenalectomy and glucocorticoid administration on the development of rat cerebral cortex. AB - In order to determine the incidence of maternal glucocorticoids on morphological parameters in fetal development, we performed optic and electron microscopic analysis of the cerebral cortex of fetuses of 16 and 20 days of gestation, from control (C) and pregnant rats bilaterally adrenalectomized on day 1 of gestation (ADX). We also studied fetuses 20 days old from pregnant rats betamethasone injected on days 15, 16 and 17 (BET), and adrenalectomized on day 1 and betamethasone-injected on days 15, 16 and 17 (ADX+BET). Absence of maternal glucocorticoids during gestation caused, in fetuses 16 and 20 days old, a marked increase of cellular density, laxity of tissue and lower cellular maturation in comparison with the control group. Beta-methasone injected into sham-operated animals (BET) caused a slight advance in relation to controls in developmental parameters such as cellular density, maturation and synapse formation. Betamethasone injection into adrenalectomized animals prevented the lower degree of maturation characteristic of the adrenalectomized group, although an increase of cellular density could be detected. The cerebral cortex from fetuses of 16 days of gestation from adrenalectomized mothers also showed an increase of cellular density as compared with the control group. These results show that glucocorticoids participate in prenatal rat brain in control mechanisms of cellular division and maturation. PMID- 7486005 TI - A study of intercellular relationships between trabecular bone and marrow stromal cells in the murine femoral metaphysis. AB - The cellular relationship between the substantia spongiosa of bone (cancellous or trabecular bone) and the haematopoietic bone marrow in the femoral metaphysis of C57BL/6NJCL mice was studied by transmission electron microscopy (TEM). Special attention was directed to intercellular junctions between osteocytes, osteoblasts, and bone marrow reticular cells. These were gap junctions and adhesive devices of simple architecture referred to as primitive junctions or zonula adherens-like junctions. Gap junctions were observed between osteocytes (within the trabeculae) and osteoblasts (at the trabecular surface) and between osteoblasts and marrow reticular cells. Gap junctions were also observed between the same cell type within each of these categories. These junctions involved the plasmalemmal membranes of adjacent cell bodies and of processes. Primitive cell junctions had a similar cellular distribution. Quantitative analysis of the cell types covering or positioned around the trabecular bones and of gap junctions between these and other cells was carried out by TEM. It was found that osteoblasts were the most numerous cell type, occupying 31% of the total of each cell type positively identified around the trabeculae (31%), while pre osteoblasts, (flattened bone marrow reticular cells) took up 26%. These data emphasise the intimate relationship of the various mesenchymal cells based on processes and intercellular junctions, and point to an anatomical and probably functional integration of trabeculae and marrow. The functional significance and putative regulatory activity of this unit are discussed. PMID- 7486006 TI - Changes in glycoconjugates revealed by lectin staining and stage-specific embryonic antigen-1 immunostaining in hamster submandibular glands during the postnatal period. AB - Lectin binding and stage-specific embryonic antigen-1 (SSEA-1) immunoreactivity were studied in the developing submandibular glands of young Syrian golden hamsters (Mesocricetus auratus) from postnatal day 1 (the day of birth) to day 28. The submandibular glands were fixed in a solution containing 6% mercuric chloride, 1% sodium acetate, and 0.1% glutaraldehyde (HgCl2-G) or 4% paraformaldehyde (4P), and embedded in paraffin. Sections from HgCl2-G fixation were stained with three lectin-peroxidase conjugates: peanut agglutinin (PNA), Ulex europeus I agglutinin (UEA I), and wheat germ agglutinin (WGA). Sections from the 4P-fixed tissues were immunostained with monoclonal antibodies against SSEA-1, sialyl SSEA-1 and fucosyl SSEA-1. On the day of birth, the terminal unit of the submandibular gland was composed of fetal type secretory cells and proacinar cells. The secretory cells were PNA, UEA I, and WGA positive. The number of secretory terminal tubule cells decreased rapidly, and lectin-positive secretory cells were replaced by adult secretory cells that did not show PNA or UEA I stainings but were weakly positive for WGA. Fetal secretory cells were positively immunostained for SSEA-1 and sialyl SSEA-1, and immature ductal cells were stained for fucosyl SSEA-1. The positive stainings disappeared with regression of the fetal epithelial cells. Hence, modulation of glycoconjugate expression in the submandibular glands, which reflects changes in secretory cells from the fetal type to adult type during postnatal development, is revealed by lectin staining and immunostaining for SSEA-1 and related antigens. PMID- 7486007 TI - Development of substance P immunoreactivity in the mouse vomeronasal organ. AB - We investigated the development of substance P immunoreactivity in mouse vomeronasal organs in embryos, juveniles, and adults. In all stages, substance P fibers were found in the receptor-free epithelial area, but never in the neuroepithelium. Substance P fibers were found sparsely in the lamina propria of 15-day-old embryos. Although buds of the vomeronasal glands in the cavernous tissue were observed in 17-day-old embryos, and gradually grew in size and numbers, the substance P fibers around them decreased after about the 13th day. Thus, substance P may be a trophic factor for the development of the vomeronasal glands in the cavernous tissue. We first recognized substance P fibers reaching the surface of the receptor-free epithelium in 13-day-old pups. In 21-day-old mice, substance P fibers were as well developed as in adult mice. Considering the development of the substance P fibers in the receptor-free epithelium and the cavernous tissue, they probably cause the vasodilation of the cavernous tissue via local axon reflexes. These structures may then act as a defense system, eliminating noxious stimulus substances sucked into the vomeronasal organ. PMID- 7486008 TI - Detection of immunoreactive atrial and brain natriuretic peptides in the equine atrium. AB - The distribution of immunoreactivity (IR) for cardiodilatin/atrial natriuretic peptide (CDD/ANP) and brain natriuretic peptide (BNP) was examined immunohistochemically and immuno-electron-microscopically in the equine atrium, using specific antibodies. In the immunohistochemical studies, IR-CDD/ANP and IR pBNP-26 (porcine BNP-26 immunoreactivity) was detected in the cytoplasm of the auricular cardiocytes, but IR-hBNP-32 (human BNP-32 immunoreactivity) was not. The double immunogold labelling method for IR-hBNP-28 and IR-pBNP-26 revealed that gold particles of different sizes were located in the same secretory granules in the cardiocyte, but no gold particles for IR-hBNP-32 were detected. These results show that CDD/ANP and porcine BNP-like peptides are colocalized in the same secretory granules of the equine atrium. They suggest that the equine atrium secretes both CDD/ANP and BNP-like peptides. PMID- 7486009 TI - Alpha calcium/calmodulin-dependent protein kinase II immunoreactivity in corticospinal neurons: combination of axonal transport method and immunofluorescence. AB - A combination of either retrograde or anterograde fluorescent tracer and immunofluorescence histochemistry using the monoclonal antibody specific for the alpha isoform of calcium/calmodulin-dependent protein kinase II (CaM kinase II alpha) was employed to test whether CaM kinase II alpha is expressed in somata of corticospinal neurons and their axons over their whole course. After the injection of carbocyanine dye DiI into the hindlimb area of the primary motor cortex of the rat, corticospinal axons and their terminal arbors were anterogradely labeled: DiI-labeled corticospinal fibers proceeded caudally in the ipsilateral internal capsule, cerebral peduncle and medullary pyramid, crossed at the pyramidal decussation and descended in the ventralmost area of the contralateral dorsal funiculus of the spinal cord. These DiI-labeled corticospinal axons expressed strong CaM kinase II alpha immunoreactivity along their course. However, their terminal arbors within the gray matter of the lumbar cord were very weakly immunostained. With the injection of Fast Blue into the lumbar enlargement of the rat, somata of corticospinal neurons in layer V of the motor cortex were retrogradely labeled. The subsequent immunofluorescent histochemistry revealed that more than 80% of Fast Blue-labeled corticospinal neurons were immunostained with CaM kinase II alpha antibody. The present immunohistochemical study demonstrated that CaM kinase II alpha is strongly expressed in both somata and axons of a majority of corticospinal neurons, although we could not detect this enzyme in the corticospinal terminals in the spinal target areas. PMID- 7486010 TI - An immunocytochemical marker for early type I muscle fibers in the developing rat hindlimb. AB - Muscle fibers develop sequentially from several generations of myotubes that express specific isoforms of myosin heavy chain (MHC). We observed that the chicken-derived monoclonal antibody (mAb) S46 binds to myotubes of the fetal rat hindlimb in a specific temporal and spatial pattern. To determine the type and fate of the S46-reactive myotubes, we immunoreacted sections of fetal, neonatal and postnatal hindlimb muscles to this antibody. The mAb S46 bound to a subpopulation of primary myotubes in the tibialis anterior, and to all primary and slow/fast secondary myotubes in the soleus muscle. The S46-reactive primary myotubes represented the oldest set of myotubes in the muscles. Reactivity to S46 was present from the earliest stages of muscle development, peaked in the late fetal period, and dissipated in the first postnatal week, suggesting that mAb S46 binds to a developmental form of slow myosin. The regional distribution of myotubes that bound S46 in fetal muscles was identical to the distribution of type I (slow-twitch) fibers in the adult, indicating that S46-reactive myotubes ultimately develop into type I extrafusal fibers. Thus, mAb S46 can be used as a marker for prospective type I extrafusal fibers in the rat hindlimb. PMID- 7486011 TI - Origin of intrafusal fibers from a subset of primary myotubes in the rat. AB - S46, a monoclonal antibody (mAb) specific for the SM-1 and SM-2 isoforms of avian slow myosin heavy chains (MHC), was used to study the earliest stages of development of intrafusal fibers in muscle spindles of the rat hindlimb. Spindles formed only in the regions of fetal muscles that contained primary myotubes reactive to mAb S46, such as the axial region of the tibialis anterior muscle. The first intrafusal fiber to form, the nuclear bag2 fiber, originated from within the population of S46-reactive primary myotubes. Binding of mAb S46 by myotubes giving rise to the bag2 fibers preceded the appearance of encapsulated spindles in the muscles by electron microscopy. However, reactivity to S46 intensified in the myotubes transforming into bag2 fibers after the innervation of the fibers by afferents, and dissipated in myotubes differentiating into slow twitch (type I) extrafusal fibers. Thus, afferents may enhance intrafusal expression of the MHC isoform reactive to mAb S46. The pattern of S46 binding to nuclear bag and chain intrafusal fibers in both developing and adult spindles was the same as that reported for the mAb ALD19, suggesting that both antibodies bind to the same MHC isoform. This isoform is probably a developmental form of slow myosin, because it was transiently expressed during the development of type I extrafusal fibers. The origin of bag2 intrafusal and type I extrafusal fibers from a bipotential subpopulation of primary myotubes reactive to mAb S46 correlates with the location of muscle spindles in the slow regions of muscles in adult rat hindlimbs. PMID- 7486012 TI - Signs of the principle body axes prior to primitive streak formation in the rabbit embryo. AB - An early common element during anterior-posterior axis formation amongst amniotes is the primitive streak, running longitudinally in the two-layered embryonic disc. In mammals the primordium of this transient structure is the first definite morphological sign of the anterior-posterior axis, while in avian embryos the axis is visible and apparently defined earlier. Here we scrutinize suggestions that in mammals also there are earlier signs of axis formation by using correlative low- and high-resolution light microscopy on tissues from rabbit embryos at 6.3 and 6.5 days post-conception, i.e. immediately before and after primitive streak formation. A series of semithin sections were cut from resin embedded embryonic discs that had been photographed previously at low power. In embryos at 6.5-days post-conception the primitive streak is as long as up to half the diameter of the embryonic disc, extending anteriorly from a thickening, here called the posterior node, at the posterior margin, which contains the first mesoderm cells ingressing from the epiblast. On both sides of the primitive streak there is a triangular area that appears light in surface views of fixed embryos and correlates with stretches of low-columnar simple epithelium in an otherwise high-columnar pseudostratified epiblast. Within the anterior margin, which has a sharper contour than the rest of the circumference of the embryonic disc, there is a narrow, crescent-shaped dark zone caused by increased cellular height and number in both epiblast and hypoblast. These characteristics of the anterior margin are also found at 6.3 days post-conception, at which stage there is no sign of a primitive streak or a posterior node. The posterior margin, in contrast, is ill-defined in these earlier embryos, or there is a light crescent within the posterior margin, which has the same histological characteristics as the bilateral posterior triangular areas of primitive streak stages. Because the anterior differentiation occurs prior to primitive streak formation and is a sign of both the anterior-posterior and the transverse axes of the embryonic disc, and because some of its histological characteristics are found in primate and human embryos, we propose to name this structure the 'anterior marginal crescent' and to add it to the list of transient structures that gradually establish the principal body axes in mammals. The anterior manifestation of body axes in mammals is thus essentially different from axis development in the avian embryo, where differentiation of these axes is first manifest at the posterior margin. PMID- 7486014 TI - Ventral ectodermal ridge and ventral ectodermal groove: two distinct morphological features in the developing rat embryo tail. AB - The ventral ectodermal ridge (VER) is a thickening of the surface ectoderm on the ventral side of the embryonic tail which resembles the apical ectodermal ridge of the limb bud. The morphological characteristics of the ventral part of the embryo tail were investigated in 10.5- to 14-day rat embryos by light microscopy of serial semithin sections and by scanning and transmission electron microscopy. In 10.5- to 11.5-day embryos the thickening of the ventral surface ectoderm includes the complete ventral midline of the tail and can be divided into two parts. The posterior part is elevated and represents the ventral ectodermal ridge. The anterior part is, in contrast to the ridge, concave, and we have termed it the ventral ectodermal groove (VEG). The cloacal membrane is located at its anterior end. Contacts between the VER and the mesenchymal cells are visible until an intact basal lamina is formed at 11.5 days. Similarly, the VEG is connected by elongated cell processes with the ventral part of the tail gut. Gap junctions are present between the apical parts of ridge and groove cells. The VEG flattens and disappears in 12-day embryos. At this stage the ridge is at its maximum height, simultaneously undergoing extensive cell death. The VER is no longer visible in 14-day rat embryos. PMID- 7486015 TI - The epithelium of canine palatine tonsils. AB - The palatine tonsil is positioned to play a key role in protecting the body against ingested microorganisms. These microorganisms must traverse the tonsillar epithelium to initiate immune reactions, but no information is available from dogs on the structure of this epithelium. In this study, the morphology and ultrastructure of the epithelium of the palatine tonsil of dogs was examined using light and electron microscopy. The epithelium is of two types: reticular and non-reticular. Reticular epithelium, which is invaded by lymphoid cells, is located over the apices of nodules, separated indistinctly by islands of non reticular squamous epithelium. The reticular epithelium contains M cells which are more abundant towards the periphery of lymphoid nodules. The apical membrane of the M cells forms folds from which microvilli extend into the lumen of the oropharynx. Abluminally, cytoplasmic processes enfold clusters of lymphocytes. Numerous desmosomes secure the marginal plasma membrane and lateral membranous interdigitations to adjacent epithelial cells, thereby maintaining the integrity of the tonsillar epithelium. Epithelial discontinuities, which are few, occupy similar positions to M cells and appear to contain migrating lymphocytes. A fenestrated basement membrane allows cell transport between the intraepithelial passageways of the reticular epithelium and the subepithelial lymphoid tissues. PMID- 7486013 TI - Lymphoid tissues of the ileum in young horses: distribution, structure, and epithelium. AB - Lymphoid tissues in the ileum of young horses form raised plaques that are macroscopically visible from the mucosal surface. These are termed "ileal lymphoid patches". These patches are variable in size, shape and position within the ileal wall, occasionally lying along the site of mesenteric attachment. Within lymphoid patches, follicles exist in three different morphological forms: follicle/dome structures, proprial follicles, and lymphoglandular complexes (LGCs). In follicle/dome structures, the majority of the follicle lies in the submucosa and merges with a dome in the lamina propria through a gap in the muscularis mucosae. In proprial follicles, the majority, or all, of the follicle is found in the lamina propria, and in LGCs, the follicles lie in the submucosa and communicate with the intestinal lumen via a central invagination of epithelium that extends vertically through a gap in the muscularis mucosae. Follicle-associated epithelium covers the follicle/dome structures and proprial follicles. It consists of enterocytes, cells morphologically resembling M cells, intraepithelial lymphocytes, goblet cells, and amine-precursor uptake and decarboxylation (APUD) cells. The epithelium of LGCs is mainly populated by immature enterocytes, intraepithelial lymphocytes and goblet cells. Cells with coarse, long microvilli are also present. Information regarding the presence of LGCs in the small intestine is scant, but LGCs have been well described in the large intestine of many species. Further investigation will be required to determine if factors exist that are common to both the ileum of the horse and the large intestine of other species to influence the development of LGCs at these specific sites. PMID- 7486016 TI - DNA changes involved in the formation of metaphase chromosomes, as observed in mouse duodenal crypt cells stained by osmium-ammine. I. New structures arise during the S phase and condense at prophase into "chromomeres," which fuse at prometaphase into mitotic chromosomes. AB - BACKGROUND: In the hope of understanding how chromosomes condense at mitosis, we took advantage of a subdivision of the cell cycle into 11 stages to examine the changes in DNA taking place during the stages preceding the emergence of metaphase chromosomes. METHODS: To identify DNA changes, pieces of mouse duodenum were fixed in formaldehyde, and sections of the rapidly dividing cells of the crypts were stained by the osmium-ammine method, which is specific for the detection of DNA in the electron microscope. RESULTS: Throughout the cell cycle, DNA is present in nucleofilaments composed of rows of 11-nm-wide nucleosomes. At stage I, during which the DNA-synthesizing or S phase of the cell cycle begins, some of the nucleofilaments are compacted in the heterochromatin accumulations associated with nuclear envelope and nucleoli, while the others are scattered in the nucleoplasm where they appear either "free" or "attached" to the heterochromatin. This DNA distribution is similar to that observed in the noncycling cells examined. After the beginning of the S phase, "free" nucleofilaments are seen to assemble into structures composed of compacted nucleofilaments and referred to as "aggregates"; these make their appearance at stage II and increase in size through stage III up to the end of S during stage IV. Meanwhile, the heterochromatin associated with nuclear envelope and nucleoli expands toward the nucleoplasm in the form of protrusions referred to as "bulges," which gradually enlarge during stages III and IV, while the heterochromatin shrinks and eventually vanishes. On average, a total of 1,171 aggregates and bulges are formed in the nucleus during the S phase. At the apparition of stage V, which corresponds approximately to prophase, aggregates and bulges are rapidly gathered into an average of 288 spheroidal bodies referred to as "chromomeres." These are connected to one another by nucleofilamentous bridges in such a way as to be lined up in rows. The formation of rows of chromomeres represents in the electron microscope the prophasic condensation observed in the light microscope. Finally, during stage VIa, which corresponds to prometaphase, the chromomeres approach one another within each row, make contact, and coalesce to become the 40 chromosomes of the mouse, which during stage VIb are organized in the equatorial plate of metaphase. CONCLUSIONS: The condensation of metaphase chromosomes occurs in three main steps. The first and longest takes place during the S phase, as nucleofilaments are assembled into aggregates, while the heterochromatin gives rise to bulges. The brief second step occurs toward the beginning of prophase, when the numerous aggregates and bulges are congregated into a limited number of chromomeres, which are lined up in rows. The third step takes place during the brief prometaphase, when the chromomeres of a row coalesce into a mitotic chromosome. PMID- 7486017 TI - DNA changes involved in the formation of metaphase chromosomes, as observed in mouse duodenal crypt cells stained by osmium-ammine. II. Tracing nascent DNA by bromodeoxyuridine into structures arising during the S phase. AB - BACKGROUND: Since it has been found that new chromatin structures make their appearance in the nucleus during the DNA-synthesizing or S phase of the cell cycle, the question arises as to how these structures are related to the nascent DNA. METHODS: DNA-containing structures were detected in sections of mouse duodenal crypt cells by the DNA-specific osmium-ammine procedure. In the same sections, the nascent or newly-replicated DNA was localized during stages I-IV of the cell cycle (corresponding to four successive parts of the S phase) by immunogold labeling of the DNA precursor bromodeoxyuridine (BrdU) in mice sacrificed 10 min after its injection. Moreover, the fate of the nascent DNA with time was traced up to 6 hr after the injection. (The nomenclature of the DNA containing structures is that proposed by El-Alfy et al., 1995.) RESULTS: Ten minutes after BrdU injection, the gold particles indicative of nascent DNA are associated with discrete nucleofilaments scattered in the nucleoplasm, but not with the compacted nucleofilaments making up the heterochromatin or the new S phase structures named "aggregates." The gold-particle-associated discrete nucleofilaments are classified into three types: a) The "free" nucleofilaments have been given this name, since they appear to be independent of heterochromatin and aggregates; nearly all gold particles are over these at stage I; but the numbers of particles over them decreases from stage I to IV. b) The "aggregate attached" nucleofilaments project from the surface of the aggregates; the number of particles over these is high at stages II and III but decreases at stage IV. c) The "heterochromatin-attached" nucleofilaments project from the surface of the heterochromatin; the number of particles over these increases from stage II to IV. By 1 hr after BrdU injection, gold particles can be over loose clumps of nucleofilaments at stages I and II, but are mostly over small aggregates at stage II, midsized aggregates and small heterochromatin-associated "bulges" at stage III and large aggregates and large bulges at stage IV. By 2-6 hr, virtually all particles are over aggregates and bulges, frequently deep within them. CONCLUSIONS: The distribution of the gold particles at 10 min reveals that DNA is synthesized in discrete nucleofilaments that are "free" or "aggregate-attached" or "heterochromatin-attached." In contrast, by one and especially two hours, the gold particles are present over aggregates and bulges, indicating that, after discrete nucleofilaments acquire nascent DNA, they are displaced to become part of these structures. More precisely, the aggregates arise from the repeated addition of replicated portions of "free" nucleofilaments, while the bulges arise from the repeated addition of replicated portions, of "heterochromatin-attached" nucleofilaments. Aggregates and bulges are the two initial building stones from which mitotic chromosomes are eventually formed. PMID- 7486019 TI - Morphology, histochemistry, and differentiation of the cat's epiglottic cartilage: a supporting organ composed of elastic cartilage, fibrous cartilage, myxoid tissue, and fat tissue. AB - BACKGROUND: In carnivores, the supporting organ of the epiglottis is usually called "epiglottic cartilage" (EC) although it is composed of elastic cartilage and unilocular fat storing cells. We studied the cat's EC in order to decide whether these fat storing cells are true adipocytes or fat storing (dedifferentiated) chondrocytes. METHODS: ECs were studied in cat embryos at gestation days 40 and 60, in newborn, postnatal, and adult cats. We used classical staining methods, immunohistochemistry, and transmission electron microscopy to identify the different kinds of tissues contributing to the EC and to follow their differentiation. RESULTS: The cat's EC was defined by a layer of coarse collagen fibers representing a tunica albuginea. This tunica covered irregularly formed and irregularly sized areas of elastic cartilage, fibrous cartilage, myxoid tissue, and lobules of unilocular fat cells. All these tissue showed regular morphology. Adipocytes were provided with continuous basal laminae and fat lobules were well supplied with capillaries. Alcianophilia of ground substance was observed in all tissue components but was strongest in elastic cartilage. Most islets of elastic cartilage adhered to the tunica albuginea of the EC at one surface and were connected to the opposite surface by coarse strands of connective tissue traversing the organ. Intercalated areas of fibrous cartilage contained fuchsinophilic collagen bundles. Myxoid tissue was characterized by stellate cells in alcianophilic ground substance with intermingled fuchsinophilic bundles. All kinds of supporting tissues combined with each other without clear demarcation. Immunohistochemistry revealed strong reactivity for S-100 of chondrocytes, myxoid cells, and fat cells. Chondrocytes and myxoid cells also stained for glial fibrillary acidic protein, neurofilament protein 200, and neuron specific enolase. During development, condensation of mesenchymal cells indicated the blastema of the EC at gestation day 40. At day 60, delicate collagen fibrils indicated the future tunica albuginea, faint alcianophilia was noted in the ground substance, and multilocular fat cells were scattered throughout the blastema. At birth, alcianophilia was moderate and multilocular fat cells were numerous. Three weeks after birth, single and grouped unilocular fat cells were seen, alcianophilia of ground substance was prominent, and former blastema cells presented as ramified myxoid cells. Eight weeks after birth, the EC primarily consisted of myxoid tissue, but the first islets of cartilage were seen in the center of myxoid areas. Unilocular fat cells already formed lobules. CONCLUSIONS: These results show that in the cat EC a) differentiation of adipocytes precedes differentiation of all the other tissue components, and b) differentiation of myxoid tissue precedes differentiation of cartilage. It is concluded that myxoid tissue may serve as a precursor of fibrous and elastic cartilage. PMID- 7486020 TI - Differences in distribution of myofiber types between the supraspinatus and infraspinatus muscles of sheep. AB - BACKGROUND: The m. supraspinatus stabilizes the shoulder joint to bear the body weight, and the m. infraspinatus assists in extension and flexion of the joint in sheep. Postural muscles have many SO myofibers, whereas locomotory muscles have numerous fast-twitch myofibers. In sheep the distribution of myofiber types within the two muscles, necessary for a better understanding of postural function, remains to be clarified. METHODS: Muscle samples were removed from the whole transverse sections of the dorsal, middle, and ventral compartments of the m. supraspinatus and m. infraspinatus of sheep. Myofibers were classified into FG, FOG, SO-1, and SO-2 myofibers by histochemical methods. RESULTS: The distribution of SO myofibers changed more greatly in the m. supraspinatus (15.0 99.1%) than in the m. infraspinatus (24.5-62.3%). SO myofibers were concentrated markedly in the caudal and deep regions near the spine and fossa of the scapula in the m. supraspinatus and distributed more in the medial part than in the lateral part in the m. infraspinatus. Such changes were caused by increases in percentage of SO-2 myofibers and not SO-1 myofibers. The craniolateral regions of the m. supraspinatus and the caudolateral regions of the m. infraspinatus had many fast-twitch (FOG plus FG) myofibers suited for rapid extension and flexion of the shoulder joint. CONCLUSIONS: The m. supraspinatus has the compartmentalized, deep, and caudal regions occupied by SO myofibers, which seem to be specialized for maintenance of the joint extension. The medial region of the m. infraspinatus may assist in the joint stabilization. PMID- 7486018 TI - Myosin heavy chain expression within the tapered ends of skeletal muscle fibers. AB - BACKGROUND: The pectoralis muscle of the chicken contains fast-twitch glycolytic fibers, which during development undergo a transformation in their myosin heavy chain (MyHC) content from embryonic to a neonatal to an adult isoform (Bandman et al., 1990). Little, however, is known of MyHC expression within the ends of these or other muscle fibers. Here we test the hypothesis that the tapered ends of mature skeletal muscle fibers contain a less mature MyHC isoform than that typically found throughout their lengths. METHODS: We apply an ammoniacal silver histological stain for endomysium and monoclonal antibodies against neonatal and adult MyHCs of chicken pectoralis to transverse serial sections of pectoralis from five mature chickens. The "lesser fiber diameters" of populations of fibers from each bird are also measured. RESULTS: Most (approximately 81.8%) of the small (< 12 microns) and none of the larger (> 20 microns) diameter fibers contain the neonatal MyHC. Following these smaller fibers through serial sections, we show that they are the tapered ends of the larger fibers. Whereas neonatal MyHC is restricted to the tapered fiber ends, adult MyHC is present throughout the entire lengths of all fibers. We also demonstrate acetylcholinesterase (AChE) activity at some of these fiber ends. CONCLUSIONS: We postulate that longitudinal growth of myofibrils in adult muscle is characterized by the sequential expression of MyHC isoforms similar to that observed in rapidly growing muscle and that the presence of the neurotransmitter hydrolase AChE at the tapered fiber ends may be related to the retention of neonatal MyHC. PMID- 7486021 TI - Kidney of elephants. AB - BACKGROUND: Elephants are an important and isolated order. Their kidneys need substantial investigation and hitherto have not been portrayed even by a pyelogram. METHODS: Pyelograms and injection of vessels with colored acrylic emulsions were done initially. Dissection was under fiberoptics using a dissecting microscope with frequent measurements. Special areas were cut for microscopy (light and electron) and photography. Glomerular counts were done by macerating weighted pieces of cortex and later finding the cortical fraction of the renal parenchyma. RESULTS: The elephant kidney is devoid of dorsoventral symmetry. It is composed of 8 +/- 2 lobes separated by fine interlobar septa. There is no reduction of lobes with maturity. The pelvis bifurcates at the sinus into primary branches or infundibula which dispatch a secondary branch or infundibulum into every lobe. Interlobar arteries and veins, nerves, fat, and connective tissue generally accompany every secondary infundibulum into its lobe. A major branch of the renal artery may perforate the renal capsule and course to the cortico-medullary (C-M) border independently of the secondary infundibulum to that lobe. The number of glomeruli per kidney is approximately 15 x 10(6). In adults the glomerular mass is 4.9 +/- 0.5% of the renal parenchyma and 6.7 +/- 0.3% of the cortex. Areae cribrosae occur generally at low papillae. They are the outlets of numerous terminal collecting ducts which may be accompanied by a tubus maximus (T.M.) A T.M. of diameter 1.6 mm and length 10 mm may act as the only substitute for an area cribrosa. Wide anastomoses between the two main renal veins occur within the renal sinus. Intralobar arteries and veins often course right through the outer medulla to and from, respectively, the C-M border. CONCLUSIONS: Anatomically, an elephant's kidneys appear to be able to concentrate urine only moderately. Their kidneys tend to resemble those of the manatee but not of the dugong. PMID- 7486022 TI - Structural features and functions of principal cells of the intermediate zone of the epididymis of adult rats. AB - BACKGROUND: In the present study, principal cells of the intermediate zone of the epididymis, an area situated between the initial segment and proximal caput, were observed to present morphological features distinct from those of principal cells of other regions. METHODS: The epididymides of adult rats were fixed by perfusion with glutaraldehyde and embedded in Epon. Administration of fluid phase tracers was performed in the case of several animals. Localization of anti-SGP-2 and anti immobilin antibodies in conjunction with light (LM) and electron (EM) microscope immunocytochemistry was also performed. RESULTS: In the LM and EM, the most distinctive feature of many principal cells of this zone was the presence of apically located vacuoles referred to as giant endosomes due to their large size and because they readily incorporated tracers introduced into the lumen of the epididymal duct and were acid phosphatase-negative. Giant endosomes, containing electron-dense granular patches, appeared to form by the progressive fusion of small, medium, and large endosomes. In the supranuclear region, multivesicular bodies (MVBs) and lysosomes were present. Although smaller in size than the giant endosomes, MVBs and lysosomes contained the electron-dense patches. It is suggested from morphological images that giant endosomes fragment into smaller units corresponding to MVBs which gradually transform into lysosomes. Experiments using anti-SGP-2 and anti-immobilin antibodies revealed gold particles over the Golgi apparatus and secretory vesicles (150-300 nm) of principal cells of this zone as well as the luminal contents indicative of secretion of these proteins. Interestingly, giant endosomes were also immunolabeled with both antibodies as were stereocilia, coated pits and vesicles, and endosomes of various sizes; lysosomes were minimally labeled. These results suggest that principal cells of the intermediate zone endocytose as well as secrete SGP-2 and immobilin. The internalized SGP-2 and immobilin may correspond to that secreted further upstream and that, possibly due to their short half-life and terminated function, are removed from the lumen of the duct. Principal cells of this zone secrete these proteins possibly to replenish that lost by endocytosis. CONCLUSIONS: Principal cells of the intermediate zone contain giant endosomes. The presence of such large structures suggests that the early events in endocytosis is a slower process in principal cells of this zone as compared to other regions. The fact that these cells both secrete and endocytose SGP-2 and immobilin adds to the complexity of our understanding of how principal cells function along the length of the epididymis. PMID- 7486023 TI - Pulmonary lymphatics and their spatial relationship to venous sphincters. AB - BACKGROUND: Pulmonary lymphatics are critical to clearing lung fluid. Although their structure can be shown with light and transmission electron microscopy, scanning electron microscopy of their casts can better show their number, size, shape, distribution, and degree of filling. This technique has identified four forms of lung lymphatics, but these forms have not been fully evaluated by tissue microscopy. A most important site of pulmonary edema formation, the pulmonary capillary, is just upstream from small veins which have focal, smooth muscle tufts termed venous sphincters. Because of their constricting potential, these sphincters may control lung perfusion and cause edema. METHODS: With light and transmission electron microscopy of tissue and scanning electron microscopy of casts, the lymphatic forms were explored in relation to the tissue anatomy in rats without pulmonary edema and with mild-to-moderate edema caused by extended vascular rinsing. RESULTS: The edematous lungs had increased sacculo-tubular lymphatics adjacent to the venous sphincters. These lymphatics were in the adventitial connective tissue and were partially endothelialized. As lymphatics became more tubular their endothelium became more complete. Collagen fibers traversed the lumen of these lymphatics even where endothelial cells were present and caused the lines on the surface of the lymphatic casts. Overlapping endothelial cells caused clefts on the casts. CONCLUSIONS: Scanning electron microscopy of lymphatic casts better defines their ultrastructure and shows the spatial relationship of veins and their sphincters to venous lymphatics. Sphincter contraction may influence pulmonary lymph production which could affect other aspects of regional lung perfusion. PMID- 7486024 TI - Linear gap and tight junctional assemblies between capillary endothelial cells in the eel rete mirabile. AB - BACKGROUND: Interendothelial tight junctions and gap junctions have been described in large blood vessels and in cultures of endothelium derived from large blood vessels. Transfer of microinjected small-molecular weight tracers between adjacent endothelial cells also has been demonstrated indicating the presence of gap junctional interendothelial communication. Similar transfer of tracers is evident between microvessel endothelial cells in culture and in microvessels in situ. However, gap junctions have not been detectable by electron microscopy of intact capillary systems. This may be due to limited sampling available in diffuse capillary systems and a small area of overlap between adjacent endothelial membranes. METHODS: Thin slices of the parallel, tightly packed capillary bed of the eel rete mirabile were cryofixed and prepared for conventional TEM by freeze-substitution. Other samples were freeze-fractured and replicated for examination of endothelial junctional components. RESULTS: A novel tight-gap junctional complex between rete capillary endothelial cells is described. In freeze-fracture replicas of the membrane P face, rows of gap junction subunits are flanked on either side by linear depressions representing grooves previously occupied by tight junctional strands that partition to the E face. In thin sections, the junctions appear in profile as short lengths of closely apposed membranes characteristic of gap junctions. CONCLUSIONS: The tight junctional components imply a barrier to paracellular transport across the capillary wall between the endothelial cells. The gap junctional component may provide a mechanism for communication between endothelial cells along the length of the vessel wall. PMID- 7486025 TI - Morphology of the left atrial appendage. AB - BACKGROUND: When examining the left atrial appendage by transesophageal echocardiography, differences in size and shape of the left atrial appendage are to be observed. The study was carried out with the aim of investigating the morphology of the left atrial appendage and to find associations with pathologic cardiac findings. METHODS AND RESULTS: In 220 cases (106 female, 114 male, mean age 72 +/- 13 years) a cast of the left atrial appendage was made after the post mortem examination by using synthetic resin. In 198 cases an ECG was available (sinus rhythm n = 143, atrial fibrillation n = 55). The casts were described in respect to course and ramifications of the principal axis. The casts were measured concerning orifice diameters, outline, and volume. Most frequently (42%) the course of the principal axis was angulated below 100 degrees. More than five ramifications of the principal axis were found in 56% of the casts. The volume ranged from 770-19,270 mm3 (mean 5,220 +/- 3,041). When comparing the clinical and autopsy-data of the patients with the morphology of the casts, associations could be found between the volume of the casts and atrial fibrillation (7,060 mm3 as compared to 4,645 mm3 in sinus rhythm, P < 0.01), left ventricular hypertrophy (5,740 mm3 as compared to 4,639 mm3 without hypertrophy, P < 0.01), myocardial scars (5,923 mm3 as compared to 4,891 mm3 without scars, P < 0.05), closed foramen ovale (5,515 mm3 as compared to 4,037 mm3 with patent foramen ovale, P < 0.01), and left atrial appendage thrombi (8,566 mm3 as compared to 5,027 mm3 without thrombi, P < 0.01). CONCLUSION: Left atrial appendages are formations greatly varying in volume and shape. This variability should be considered when interpreting images of the left atrial appendage, and in particular when diagnosing thrombi. PMID- 7486026 TI - Pericyte-endothelial gap junctions in developing rat cerebral capillaries: a fine structural study. AB - BACKGROUND: Fine structural study revealed the intercellular coupling between the pericyte and the endothelial cells via the gap junctions, in the capillaries of the basal forebrain of rat embryos. RESULTS: Gap junctions were constructed by the adluminal plasmalemma of pericyte and the abluminal plasmalemma of endothelial cells. CONCLUSIONS: Gap junctions are membranous channels that directly join the cytoplasms of the pericyte and endothelial cell and imply some substantial role for the pericyte on the endothelial proliferation. It is postulated that the function of the pericyte in the prenatal mammals are assigned to the regulation of the development of cerebral microcirculation. PMID- 7486027 TI - Sources of subcortical afferents to the macaque's dorsal lateral geniculate nucleus. AB - BACKGROUND: The dorsal lateral geniculate nucleus (dLGN) is the thalamic region responsible for transmitting retina signals to cortex. Brainstem pathways to this nucleus have been described in several species and are believed to control the retinocortical pathway depending on the state of the animal (awake, asleep, drowsy, etc.). The purpose of this study was to determine all of the subcortical sources of afferents to the dLGN in a higher primate, the macaque monkey, whose visual system is similar to that of humans. METHODS: Injections of horseradish peroxidase (HRP), with or without conjugation to wheat germ agglutinin, were made into the dLGNs of seven macaque monkeys, followed by perfusion, brain sectioning, and analyses of neurons in the brainstem, thalamus, and hypothalamus that contained the retrogradely transported marker. RESULTS: The reticular nucleus of the thalamus, pedunculopontine nucleus, parabigeminal nucleus, pretectal nucleus of the optic tract, superior colliculus, dorsal raphe nucleus, and tuberomammillary region of the hypothalamus contained many retrogradely labeled neurons ipsilateral to the injections. In the contralateral brainstem, HRP labeled cells were found only in the pedunculopontine nucleus, nucleus of the optic tract, and dorsal raphe nucleus. The number of labeled neurons on the contralateral side was about one-half of that in corresponding ipsilateral nuclei. The locus coeruleus contained no labeled neurons in four of the macaques that had injections limited to the dLGN. CONCLUSION: There are seven subcortical regions that send afferents to the dLGNs of macaque monkeys. Except for the locus coeruleus, these are the same as observed for other species, such as the cat and rat, and indicate the possible sources of subcortical control over the dLGNs of humans. PMID- 7486028 TI - Patella of selected bats: patterns of occurrence or absence and associated modifications of the quadriceps femoris tendon. AB - BACKGROUND: Having observed the apparent absence of a bony patella in a Madagascar flying fox (Pteropus sp.), other species from the two suborders of bats (Megachiroptera and Microchiroptera) were examined to determine the presence or absence of a bony patella and the distribution of this feature among bats. METHODS: Gross, radiographic, and histologic examination of seven megachiropteran species representing four genera, as well as six microchiropteran species representing six genera, was performed. RESULTS: A bony patella was observed in all six microchiropteran and in three megachiropteran species. The tendon of the quadriceps femoris muscle in Microchiropteran species was composed mainly of dense regular connective tissue. The quadriceps tendon in Megachiropteran species with a patella contained an abundance of fibrocartilage and hyaline cartilage, unlike the quadriceps femoris tendon of the Microchiroptera or a laboratory mouse examined for comparison. CONCLUSIONS: Four species of the megachiropteran genus Pteropus lacking a bony patella displayed a similar occurrence and distribution of fibrocartilage and hyaline cartilage within the quadriceps tendon as seen in the other bats. In reference to this singular feature, Pteropus is unique among the representatives of megachiropteran and microchiropteran genera examined here. PMID- 7486029 TI - The apical mitochondria-rich cells of the mammalian epididymis. AB - The morphology and function of the apical mitochondria-rich cells in the mammalian ductus epididymidis epithelium are revised. These cells are similar in all mammalian species studied. Apical mitochondria-rich cells are scarce (1-5 cells/100 principal cells) and are mainly found in the initial epididymal segments. Their morphology varies from slender cells that extend from the basal lamina to the epididymal lumen, to round cells that protrude into the lumen and are not in contact with the basal lamina. Their cytoplasm is more electron-dense than that of principal cells and contains more mitochondria which, in some species, are surrounded by rough endoplasmic reticulum cisternae. The adluminal cytoplasm displays a few short microvilli and contains many acid phosphatase positive vesicles. Apical mitochondria-rich cells differ from the principal cells in some histochemical features such as: (a) different lectin-staining pattern; (b) more intense reaction to the enzymatic activities: carbonic anhydrase, Ca(2+) ATPase, peanut-agglutinin-sialidase, NADP dehydrogenase, succinate dehydrogenase, alpha-galactosidase and beta-galactosidase; (c) more intense immunoreaction to several cytokeratin types and to estradiol-related receptor protein; (d) weaker immunoreaction to epithelial membrane antigen and to retinol-binding protein. Although the function of the apical mitochondria-rich cells is still unknown, the following possible functions have been suggested: holocrine secretion; cooperation with the principal cells in epididymal reabsorption of testicular fluid; and acidification of epididymal fluid. Experimental results suggest that differentiation and maintenance of apical mitochondria-rich cells are not under androgen control and that these cells are sensitive to estrogen stimulation. PMID- 7486030 TI - Endocrine and spermatological characteristics of 135 patients with bilateral megalotestis. AB - Little is known about the biological and clinical significance of bilaterally enlarged testes (bilateral megalotestes or macroorchidism). We have investigated clinical, endocrine and spermatological characteristics of 135 andrological patients with a testicular volume of more than 25 ml bilaterally. 135 patients with normal testicular volumes served as controls. Subjects with bilateral megalotestis had higher sperm counts and better sperm morphology and motility than the controls. Levels of LH, FSH, prolactin, testosterone and estradiol were lower in the megalotestis group. This was statistically significant for LH and FSH. No testicular tumours were diagnosed in either group. We conclude that in most andrological patients with bilateral megalotestes fertility parameters are excellent despite the unusually low hormone levels found in a significant minority. No specific pathology underlying the large gonadal volume could be identified. PMID- 7486031 TI - Sympathetic auto-immune orchitis. AB - There is increasing evidence that, besides the immunization to sperm antigens and other immunological disorders, spontaneous autoimmune orchitis may also be an aetiological factor in male reproductive failure. The present paper describes a case study, where a surgical operation for an inguinal hernia caused an orchitis first in the ipsilateral, and few weeks later also in the contralateral testis and which finally led to an atrophy of both testicles. The time-course, patient's symptoms and especially the final histological picture all indicate that this process had an immunological basis, and therefore it is proposed that it should be called 'auto-immune sympathetic orchitis'. PMID- 7486032 TI - The effective cumulative pregnancy rate of different modes of treatment of male infertility. AB - The clinical efficacy of conventional and advanced methods of treatment was assessed in 814 couples with infertility due to a male factor. The monthly and effective cumulative rate of ongoing or term pregnancies was calculated during 4712 couple-months. Treatment of varicocele by transcatheter embolization, resulting in 3.9% pregnancies per cycle and an effective cumulative pregnancy rate of 41% after 1 year, is more effective than counselling and timed intercourse (9% pregnancies after 12 months). Intrauterine insemination (IUI) of washed spermatozoa produced 17% pregnancies in the initial 4 months, but the success rate of the subsequent cycles (1.7% per cycle) was not different from that of the controls. In vitro fertilization (IVF) resulted in 16% pregnancies per attempt, but the effective cumulative pregnancy rate was only 31% in 12 months due to the long interval between treatment attempts and the high drop-out rate. With subzonal microinjection of sperm, the fertilization rate was higher (71%) than with regular IVF (29%) but both the pregnancy rate per attempt (9%) and the effective cumulative pregnancy rate (17% after 12 months) were low. The 10th percentile of sperm characteristics (cut-off values) of successful cases showed intrauterine insemination to be advantageous in cases with a lower percentage of spermatozoa with progressive motility (9%) than in the controls (15%). The cut-off value of sperm morphology in IVF (4%) is lower than that of IUI (8%) and of the controls (9%), but higher than that of subzonal insemination (1%). Treatment strategy must be defined selecting or combining conventional and assisted reproductive technology for each individual couple with male factor infertility. PMID- 7486033 TI - Evolution of somatic and germ cell populations after busulfan treatment in utero or neonatal cryptorchidism in the rat. AB - In order to elucidate the respective effects of depletion of germ cells and of increase in testicular temperature, rats of the same Wistar strain were rendered experimentally bicryptorchid or sterilized by a busulfan injection in utero and compared to control animals. In both models, germ cells were depleted but numeric evolution and functions of somatic cells differed. The aim of that work was to compare the numeric evolutions of testicular somatic and germ cells to their respective functions in each model before puberty and in adult rats of the same strain. Serum concentrations of FSH, LH and testosterone were compared at 20, 40 and 110 days of age. Histological analyses of Sertoli and germ cells in the seminiferous tubules and of Leydig cells in the intertubular tissue were performed before puberty and at adulthood. Testosterone serum concentrations were depleted in both models starting at 40 days of age and more in busulfan-treated rats. Both FSH and LH levels were increased from 20 days onwards in experimental rats. Additional cryptorchidism in busulfan-treated rats depressed the serum testosterone concentration. At 17 days of age, the cryptorchidism do not modify somatic or germ cell populations while busulfan treatment has induced a decrease of both these populations. Conversely, the cross sectional area of the somatic testicular cells was not affected whatever the treatment.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7486034 TI - Effect of milk-yolk on the fertilizing capacity of spermatozoa. AB - Pre-incubation of human sperm in egg yolk based medium (TEST-yolk) as well as milk have been shown to improve sperm fertilizing capacity. The purpose of this study was to compare a combination of these two media, called milk-yolk, against the two established media, in order to test for a possible synergistic effect(s) on sperm functions as measured by sperm motility, acrosome reaction and penetration assay. Statistically, no differences have been observed among the three media, except for SPA outcome in milk-yolk, being significantly lower than the corresponding values for the other two media. In conclusion the beneficial effects of egg yolk and milk appear to be eliminated once both media are combined, resulting in an antagonistic effect on the fertilizing capacity of the spermatozoa. PMID- 7486035 TI - Vesicles in rat epididymal fluid. Existence of two populations differing in ultrastructure and enzymatic composition. AB - Glycosidase activity is very high in rat epididymal fluid as a consequence of the secretory capacity of the epithelium. The mechanism of this secretion is, so far, unknown. Membrane-bound vesicles with activity of beta-galactosidase and N-acetyl beta-D-glucosaminidase were previously isolated by us from rat epididymal fluid. We report here the existence of two populations of epididymal vesicles separated by centrifugation in a sucrose gradient. They were found to differ in isopicnic equilibrium, size, ultrastructure, and enzymatic activity. Seven days after castration the protein content and specific activities of both enzymes were found decreased in the fractions containing the vesicles. A role in enzyme secretion by the epididymal epithelium is suggested for each vesicle population. PMID- 7486036 TI - The presence of alpha-glucosidase, glycerophosphocholine and carnitine in the epididymis and ejaculate of the vervet monkey (Cercopithecus aethiops) and the Chacma baboon (Papio ursinus). AB - The epididymis is the site of posttesticular sperm maturation in the male genital tract. Studies on human epididymides are hampered by the practical inaccessibility of epididymides of healthy men in their reproductive years. The limited use of laboratory animals therefore seems unavoidable. The objective was to establish baseline values of the epididymal markers alpha-glucosidase, glycerophosphocholine (GPC) and carnitine in the lumen of the caput, corpus and cauda epididymidis and in the ejaculate of adult male Chacma baboons and vervet monkeys. In both primates, alpha-glucosidase was found throughout the epididymis and in the ejaculate; values did not vary significantly. In monkeys, the highest concentration of GPC was found in the cauda epididymidis, but smaller amounts were found in the other regions and the ejaculate. In baboons, GPC was absent from the caput, but present in the other regions, including the ejaculate. Carnitine concentrations increased significantly from the caput to the cauda in monkeys and from the caput to the corpus in baboons. With this study, the relative concentration ranges in which these markers are present in the epididymides of these primates have been established. In future studies, changes in concentrations of these substances would probably indicate changes in epididymal function. PMID- 7486037 TI - Laryngeal mask airway positioning is related to Mallampati grading in adults. AB - The Mallampati classification is a commonly used means of preoperatively predicting a difficult endotracheal intubation. As the laryngeal portion of the laryngeal mask airway (LMA) must sit over the larynx, we wondered whether the Mallampati classification also predicts difficulty in achieving adequate seating of the LMA. LMA positioning was assessed prospectively in 100 adult patients by fiberoptic bronchoscopy to determine whether there was a relationship between the ease of seating of the LMA and the Mallampati classification. In 72 patients, optimal seating of the LMA was achieved on the first attempt at insertion, and all these patients were classified as Mallampati class 1 or 2. In all 28 cases of difficulty with LMA insertion, the patients were Mallampati class 2 or 3. In two cases the LMA was abandoned, and in these cases both patients were Mallampati class 3, (P = 0.0001 by chi 2 analysis). We conclude that the Mallampati classification indicates difficulty not only in tracheal intubation but also in achieving an adequate airway with the LMA. PMID- 7486038 TI - Antagonism of mivacurium neuromuscular block: neostigmine versus edrophonium. AB - This study was designed to compare the effectiveness of antagonism of mivacurium blockade with either neostigmine, edrophonium, or spontaneous recovery. Thirty ASA physical status I or II patients provided informed consent and were randomized to one of the following groups: Group 1, placebo saline; Group 2, edrophonium (1 mg/kg); and Group 3, neostigmine (70 micrograms/kg) (n = 10/group). All studied patients had anesthesia induced with propofol and maintained with propofol/N2O/fentanyl. Mivacurium bolus of 0.2 mg/kg was used for endotracheal intubation and an infusion titrated to maintain deep levels of block (T1% = 1%-5%) (T1% = first response/control response x 100). The antagonist was injected at a deep level of the block (T1% = 1%-8%) and neuromuscular (NM) recovery was evaluated by train-of-four twitches (TOF). T1% was used during maintenance, whereas both T1% and TOF% (fourth response/first response x 100) were used during recovery. Investigators were blinded to the antagonist used. Plasma cholinesterase activity was measured prior to antagonist administration (0 min), as well as 15, 30, and 60 min after. Plasma cholinesterase activity was decreased to 29% of control at 15 min and remained at approximately 60% of the control after neostigmine administration. Edrophonium did not affect plasma cholinesterase activity. Clinically adequate spontaneous recovery (TOF% > or = 70%) of the mivacurium block with placebo required 15-18 min. On average, clinically adequate antagonism of mivacurium by edrophonium was 50% faster than placebo and 30%-40% faster than with neostigmine. In summary, the speed of antagonism with edrophonium is faster than with neostigmine when antagonizing deep mivacurium NM block.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7486039 TI - A two-center comparison of the cardiovascular effects of cisatracurium (Nimbex) and vecuronium in patients with coronary artery disease. AB - Cisatracurium (Nimbex) is an intermediate-acting benzylisoquinolinium neuromuscular blocker that is one of the stereoisomers of atracurium. It causes no clinically significant cardiovascular side effects or histamine release in doses up to 8 x ED95 in healthy patients. Seventy patients undergoing elective myocardial revascularization consented to participate in an Institutional Review Board approved pilot study (10 patients) and an open-label, randomized, controlled trial comparing the hemodynamic effects of cisatracurium with vecuronium (60 patients) at two centers. The patients were anesthetized using 100% oxygen, fentanyl, and midazolam, and tracheal intubation was facilitated with succinylcholine. At least 5 min after tracheal intubation, baseline hemodynamic measurements were obtained. The patients received 0.10 mg/kg of cisatracurium (2 x ED95) or 0.10 mg/kg of vercuronium (2 x ED90) as follows: cisatracurium over 60 s (Pilot Group A, n = 5); cisatracurium over 30 s (Pilot Group B, n = 5); cisatracurium over 5-10 s (Group C, n = 30); or vecuronium over 5-10 s (Group D, n = 30). The hemodynamic measurements were repeated at 2, 5, and 10 min after cisatracurium or vecuronium injection. There were no episodes of cutaneous flushing. One patient was hypotensive before and after cisatracurium administration, and was excluded from analysis. Otherwise, there were no episodes of hypotension requiring therapy in any patient after cisatracurium. Fifteen patients overall were excluded from the analysis for one or more of the following: light anesthesia, treatment for hypotension < 10 min prior to baseline, or equipment difficulties.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7486040 TI - The effect of ketamine or thiamylal on succinylcholine-induced myoglobinemia under halothane anesthesia in adults. AB - We studied the effect of intravenous (IV) thiamylal or ketamine on the increases in serum myoglobin, creatine kinase (CK), and potassium induced by succinylcholine under halothane anesthesia. Ninety patients were divided into three groups. In Group O, succinylcholine (1.0 mg/kg) was administered after an the inhaled induction of anesthesia with halothane. In Group T, thiamylal (4.0 5.0 mg/kg), and in Group K, ketamine (1.5-2.0 mg/kg) was injected prior to succinylcholine administration. Anesthesia was maintained with halothane, nitrous oxide, and oxygen. Myoglobin increased in Group O at 60 min to 855 ng/mL (median) and CK to 98.6 IU/L. These increases were significantly higher than those of Group T (110 ng/mL and 66.4 IU/L) and Group K (110 ng/mL and 57.0 IU/L). Potassium increased in Group O only. There was no significant difference between Groups T and K for any of the three values. These results indicate that IV thiamylal and ketamine both suppress the increases of myoglobin, CK, and potassium, and that in cases when succinylcholine is used, both drugs can help to avoid myoglobinemia and limit the increases of CK and potassium. PMID- 7486041 TI - In vitro plasma protein binding of neuromuscular blocking agents in different subpopulations of patients. AB - In vitro protein binding of several neuromuscular blocking agents (NMBAs) was measured by ultrafiltration in plasma from patients susceptible to demonstrate changes in their protein constituents. First, the relationship between the free fraction of atracurium and plasma lipoproteins levels in young volunteers (22-32 yr old, n = 6) and hyperlipidemic patients (44-68 yr old, n = 13) was studied, and second, the free fraction of atracurium, mivacurium, doxacurium, and vecuronium was determined in plasma of healthy young (27-47 yr old, n = 10), elderly (72-89 yr old, n = 11) and obese (21-57 yr old, n = 9, 200%-360% ideal body weight) patients scheduled for elective surgery. In hyperlipidemic patients, atracurium free fraction was significantly less than in young volunteers (40% +/- 5% vs 50% +/- 5%, mean +/- SD), and decreased as total cholesterol, low-density lipoprotein cholesterol and triglycerides increased (P < 0.05). In young individuals, NMBA free fraction was 25% +/- 5% for vecuronium, 58% +/- 8% for doxacurium, 52% +/- 6% for atracurium, and 72% +/- 3%, 70% +/- 3%, 70% +/- 4% for mivacurium trans-trans, cis-trans, and cis-cis isomers, respectively. Higher triglyceride concentrations in obese patients and lower high-density-lipoprotein cholesterol concentrations in both obese and elderly patients were observed when compared with young subjects. However, there was no significant difference in protein binding of NMBAs among these three groups. We conclude that, in otherwise healthy patients, age and weight are not likely to alter the free fraction of NMBAs. PMID- 7486042 TI - The comparative effects of equimolar sevoflurane and isoflurane in isolated hearts. AB - The aim of this study was to compare the direct effects of equivalent molar concentrations of sevoflurane (SEVO) and isoflurane (ISO) on electrophysiology, mechanical function, metabolism, and perfusion in isolated hearts, independent of neuronal, humoral, or hemodynamic influences. Three equimolar concentrations of SEVO or ISO were administered randomly in each of 14 guinea pig hearts perfused by the Langendorff technique. Spontaneous heart rate (HR), atrioventricular (AV) conduction time, left ventricular pressure (LVP), and coronary flow (CF) were measured directly. To differentiate a direct vasodilatory effect from an indirect metabolic effect due to autoregulation of CF, arterial and coronary sinus oxygen tension were measured continuously to calculate oxygen delivery (Do2), myocardial oxygen consumption (MVo2), percent O2 extraction, and cardiac efficiency. Linear slope analysis (cardiac effect as a function of 0.1 mM anesthetic concentration) was used to compare anesthetic effects. Only AV time was increased more (P < 0.05) by ISO (+1.8 ms per 0.1 mM) than by SEVO (+1.1 ms per 0.1 mM). CF tended to be higher with ISO (+0.7 mL.g-1.min-1 per 0.1 mM) than SEVO (0.4 mL.g-1.min-1 per 0.1 mM) but this was not significant. HR (ISO, -1.4% per 0.1 mM; SEVO, -1.7% per 0.1 mM), LVP (ISO, -5.8% per 0.1 mM; SEVO, -5.1% per 0.1 mM), and percent O2 extraction (ISO, -6.1% per 0.1 mM; SEVO, -5.8% per 0.1 mM) were decreased similarly by both anesthetics and these effects were accompanied by proportional decreases in MVo2 (ISO, -34% +/- 4%, SEVO, -37% +/- 6%) at the highest concentrations (0.53 mM). Neither anesthetic altered cardiac efficiency.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7486043 TI - Lethal toxicity from equimolar infusions of cocaine and cocaine metabolites in conscious and anesthetized rats. AB - We compared the lethal toxicity of cocaine with that of three of its metabolites to determine the contribution of these metabolites to the lethal potential from cocaine infusion. Equimolar quantities of cocaine, norcocaine, benzoylecgonine, and ecgonine methyl ester were infused in conscious rats to determine onset of convulsions and respiratory arrest. In addition, the convulsive and respiratory toxicity for cocaine and norcocaine were evaluated in anesthetized rats and their circulatory toxicity in anesthetized and ventilated rats. Norcocaine infusion resulted in earlier onset of convulsions and respiratory arrest in conscious rats than cocaine and earlier onset of circulatory arrest. Plasma concentrations of norcocaine and cocaine were not different at these times. Benzoylecgonine and ecgonine methyl ester were less potent convulsants and respiratory depressants than norcocaine and cocaine, with ecgonine methyl ester more respiratory depressant than benzoylecgonine. Pentobarbital anesthesia enhanced the respiratory depression and suppressed or delayed the onset of convulsions from norcocaine and cocaine infusion. Prolonged infusion of cocaine to circulatory arrest resulted in benzoylecgonine concentrations approximately 60%, and norcocaine concentrations approximately 5%, of the cocaine concentration, but no detectable ecgonine methyl ester formation. We conclude that although norcocaine, ecgonine methyl ester, and benzoylecgonine administered separately have lethal potential in massive dosages, death from cocaine overdose primarily results from the parent compound and not from metabolite formation. PMID- 7486045 TI - Role of minor electrolytes when applied to Stewart's acid-base approach in an acidotic rabbit model. AB - Current clinical practice regarding acid-base balance is derived from the traditional Henderson-Hasselbalch equation. An alternate theory, developed by Stewart, relates hydrogen ion concentration ([H+]) changes to three independent variables: the strong ion difference, weak acids, and PCO2. The insight into acid base balance gained by this approach has potentially important clinical applications. To test the theory in vivo, electrolyte and blood gas values were measured in rabbits subjected to a period of hypoxia sufficient to produce a severe metabolic acidosis, followed by a period of reoxygenation. These variables were used to calculate [H+] by Stewart's original formula, which was then compared to the measured value. A high correlation was found between measured and calculated values for a pH range of 7.3-7.6; however, progressive deterioration in correlation was observed outside this range. Perhaps the presence of an unidentified anion, thought to be phosphate, causes a breakdown in the accuracy of Stewart's formula at pH extremes. The addition of phosphate values to the calculation, along with calcium and magnesium, restored the agreement throughout the physiologic range. To make the formula clinically applicable, modification to incorporate phosphate levels is required to achieve the necessary accuracy in biologic solutions at more acidemic pH ranges. PMID- 7486044 TI - Creatine kinase alterations after acute malignant hyperthermia episodes and common surgical procedures. AB - Skeletal muscle may release creatine kinase (CK) during a malignant hyperthermia (MH) episode; however, muscle damaged during surgery may also release CK. This study examined the overlap between peak plasma CK levels in patients suspected of having had a MH episode (data obtained from North American MH Registry) and previously published CK changes occurring after common surgeries. For patients who were subsequently proven to be MH positive by muscle biopsy, there was considerable overlap. This was most significant with surgeries having substantial tissue damage, such as major vascular surgery and abdominal surgery. Overlap was much less with minimally invasive surgery, such as cystoscopy. Approximately 30% of MH positive patients treated with dantrolene had peak CK in the range of most surgical procedures, and approximately 50% of MH positive patients not given succinylcholine had peak CK similar to those of most surgical procedures. Dantrolene did not significantly alter peak CK in MH positive patients; however, succinylcholine was associated with significantly higher peak CK. These data suggest that patients who have had an acute MH episode during a surgical procedure may have peak CK values within the range of CK values expected from the procedure itself. PMID- 7486046 TI - Nitric oxide: physiology and pharmacology. PMID- 7486047 TI - Perioperative management of aneurysmal subarachnoid hemorrhage: Part 1. Operative management. PMID- 7486049 TI - Capnography during transtracheal needle cricothyrotomy. PMID- 7486048 TI - The incidence of complications after the double-catheter technique for cannulation of the right internal jugular vein in a university teaching hospital. PMID- 7486050 TI - A new method for difficult endotracheal intubation: the use of a jet stylet introducer and capnography. PMID- 7486052 TI - The effect of carrier intravenous fluid speed on the injection pain of propofol. PMID- 7486051 TI - A new endotracheal tube with a cuff impervious to nitrous oxide: constancy of cuff pressure and volume. PMID- 7486053 TI - Intraoperative rhabdomyolysis in a patient receiving pravastatin, a 3-hydroxy-3 methylglutaryl coenzyme A (HMG CoA) reductase inhibitor. PMID- 7486054 TI - Compartment syndrome of the forearm and hand after brachial artery cannulation. PMID- 7486056 TI - Anesthetic hazards in a previously unsuspected case of posterior mediastinal paraganglioma. PMID- 7486055 TI - Improvement of arterial oxygen saturation with selective lobar bronchial block during hemorrhage in a patient with previous contralateral lobectomy. PMID- 7486057 TI - Bronchospasm during pneumoperitoneum. PMID- 7486058 TI - Multiplane transesophageal echocardiographic guidance during resection of renal cell carcinoma extending into the inferior vena cava. PMID- 7486059 TI - Temporary facial nerve palsy during carotid endarterectomy under local anesthesia. PMID- 7486060 TI - Complication of cervical plexus block or fugue state? PMID- 7486062 TI - Addition of low-dose ketamine to general anesthesia does not improve cardiovascular response during conventional abdominal surgery. PMID- 7486064 TI - Precipitation of thiopental by some muscle relaxants. PMID- 7486063 TI - Altering epidural morphine side effects with butorphanol in children. PMID- 7486061 TI - Nonsteroidal antiinflammatory drugs, opioids, and routine anesthesia. PMID- 7486065 TI - Transesophageal echocardiography and normal variants. PMID- 7486066 TI - Which method for intraoral glossopharyngeal nerve block is better? PMID- 7486067 TI - Transverse cervical artery pseudoaneurysm: a complication of internal jugular vein catheterization. PMID- 7486068 TI - Oxygen consumption during profound intraoperative hemodilution. PMID- 7486069 TI - Antiplatelet therapy, regional anesthesia, and spinal hematomas. PMID- 7486070 TI - Use of the self-inflating bulb with the Esophageal Tracheal Combitube. PMID- 7486072 TI - Risk of pulmonary angiography. PMID- 7486071 TI - Risk factors of transient atrioventricular block after cardiac surgery. PMID- 7486073 TI - Perioperative torsade de pointes ventricular tachycardia. PMID- 7486074 TI - Cardiac anesthetic: is it unique? PMID- 7486075 TI - Cost containment: at what expense? PMID- 7486076 TI - The effects of anesthetic technique on the hemodynamic response and recovery profile in coronary revascularization patients. AB - This study was undertaken to assess the effects of propofol (versus enflurane, fentanyl, and thiopental) on hemodynamic stability and recovery characteristics when used for maintenance of anesthesia during elective coronary artery bypass grafting (CABG) procedures. Ninety premedicated patients scheduled for elective coronary revascularization had anesthesia induced with fentanyl 25 micrograms/kg intravenously (i.v.). When the mean arterial blood pressure (MAP) increased 10% above preoperative baseline values, patients were randomized to receive one of four anesthetic treatments: enflurane, 0.25-2.0%; fentanyl, 10-20 micrograms/kg i.v. bolus doses; propofol, 50-250 micrograms.kg-1.min-1 i.v.; or thiopental, 100 750 micrograms.kg-1.min-1 i.v.. The maintenance anesthesia was titrated to achieve hemodynamic stability (i.e., maintain the MAP within 10% of the baseline values and heart rate [HR] within 20% of the baseline values). After bypass, anesthetic and cardiovascular drugs were titrated to maintain the MAP > 65 mm Hg and the cardiac index (CI) > 2.3 L.min-1.m-2. Recovery was assessed by noting the times at which patients first opened their eyes, responded to verbal communication, correctly responded to specific commands, underwent tracheal extubation, and were discharged from the intensive care unit (ICU). Although less intraoperative hypertension was noted in the propofol-treated patients (19 +/- 11 min vs 38 +/- 26 min, 30 +/- 24 min, and 30 +/- 23 min in the enflurane, fentanyl, and thiopental groups, respectively) (P = 0.04), the incidence of hypotension did not differ significantly among the groups. Vasopressor drugs were required more often during the prebypass period in fentanyl and propofol patients (4/22 and 5/23, respectively) compared to the thiopental group (0/21) (P < 0.05). During CPB, fentanyl-treated patients required vasoconstrictors more often than patients in the other three treatment groups (14/22 vs 6/24, 4/23, and 5/21 in the enflurane, propofol, and thiopental groups, respectively) (P < 0.01). Although fentanyl-treated patients had significantly greater requirements for inotropic support during weaning from CPB than propofol-treated patients (14/22 vs 7/23) (P < 0.038), there were no significant differences among the groups in the postbypass or ICU periods. Propofol-treated patients responded to verbal stimuli (2.1 +/- 1.3h vs 4.0 +/- 3.5h, 4.7 +/- 2.7h, and 5.6 +/- 3.6h in the enflurane, fentanyl, and thiopental groups, respectively) (P = 0.01) and followed commands earlier (propofol 7.3 +/- 5.2h vs enflurane 12.5 +/- 5.7h, fentanyl 13.1 +/- 6.6h, and thiopental 12.8 +/- 6.7 h) (P = 0.01).(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 7486077 TI - Cerebral metabolic consequences of hypotensive challenges in hemodiluted pigs with and without cardiopulmonary bypass. AB - We tested the hypothesis that progressive aortic hypotension with bicarotid occlusion produces greater reductions in cerebral blood flow (CBF) and more flow metabolism mismatching with hemodilution during cardiopulmonary bypass (CPB) than with hemodilution alone. In Yorkshire pigs randomized to hemodilution with CPB (n = 10) or hemodilution without CPB (control; n = 9), the effects of bicarotid ligation and graded hypotension on CBF (microspheres), the electroencephalogram (EEG), and cortical energy metabolites were examined. After bicarotid ligation, systemic flow was reduced for 15-min intervals of 80, 60, and 40 mm Hg aortic pressure, followed by a cortical brain biopsy. At baseline, CBF was lower in CPB (58 +/- 3 mL.100g-1.min-1) than control (90 +/- 3 mL.100 g-1.min-1., P < 0.05) animals, as was cerebral oxygen metabolism (3.1 +/- 0.1 vs 4.2 +/- 0.2 mL.min 1.100g-1; P < 0.05). Although CBF remained 40% lower at each level of hypotension in CPB than control animals (P < 0.05), EEG scores showed no intergroup differences, indicating similar flow-metabolism matching. Brain metabolites were similar between CPB and control groups (adenosine triphosphate, 9.6 +/- 2.4 vs 12.4 +/- 1.9 mumol/g; adenosine diphosphate, 6.0 +/- 0.7 vs 6.3 +/- 0.4 mumol/g; adenosine monophosphate, 4.8 +/- 0.9 vs 3.8 +/- 0.8 mumol/g; creatine phosphate, 8.3 +/- 1.8 vs 7.9 +/- 1.0 mumol/g; and lactate, 178.4 +/- 20.2 vs 150.8 +/- 13.9 mumol/g). Thus, despite significantly lower CBF during hypotension with bicarotid occlusion in hemodiluted animals during normothermic CPB, cortical electrical activity and the balance between flow and metabolism did not differ from those in control animals without CPB. PMID- 7486078 TI - Aprotinin prolongs whole blood activated partial thromboplastin time but not whole blood prothrombin time in patients undergoing cardiac surgery. AB - Aprotinin is being used increasingly to limit cardiopulmonary bypass (CPB) induced coagulation derangements. Since whole blood prothrombin time (PT) and activated partial thromboplastin time (APTT) assays are beneficial in the treatment of bleeding after CPB, we studied the potential effect of aprotinin on these whole blood assays. Blood specimens from 151 cardiac surgical patients were obtained in two phases: prior to heparin administration, before CPB, and subsequent to heparin neutralization after CPB. After collection, blood specimens were divided into two aliquots and mixed with either normal saline (NS) or aprotinin (A, 200 or 400 Kallikrein inhibiting units (KIU)/mL). Whole blood specimens were used to measure whole blood PT and APTT using CoaguChek Plus instruments. Whole blood PT results were similar between normal saline. (NS)- and aprotinin-spiked specimens before CPB (A, 12.9 +/- 1.5s; NS, 12.8 +/- 1.5s; P = 0.76) and after CPB (A, 17.5 +/- 2.4s; NS, 17.7 +/- 2.4s; P = 0.58). In contrast, whole blood APTT results were prolonged in aprotinin-spiked specimens prior to CPB (A, 63.3 +/- 32.2s; NS, 38.6 +/- 16.3s; P < 0.0001) and after CPB (A, 65.9 +/ 23.7s; NS, 45.7 +/- 14.4s; P < 0.0001). A dose-dependent prolongation of whole blood APTT by aprotinin was demonstrated by a greater mean difference in APTT (P = 0.0001) between specimens spiked with NS or 200 KIU (17.5 +/- 12.2s) vs 400 KIU (27.8 +/- 21.5s) of aprotinin.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7486079 TI - The direct and interactive effects of phosphodiesterase inhibition and beta adrenergic stimulation on myocyte contractile function after hypothermic cardioplegic arrest. AB - The direct and interactive effects of phosphodiesterase inhibition (PDEI) and beta-adrenergic receptor (beta AR) stimulation on isolated myocyte contractile function were examined after hypothermic, hyperkalemic, cardioplegic arrest (HHCA) and under normothermic conditions. Left ventricular (LV) myocytes were isolated from porcine hearts and myocyte contractile function was measured under normothermic conditions (37 degrees C in standard media) and after HHCA (2 h at 4 degrees C in Ringer's solution with 24 mEq KCl) with subsequent rewarming. Myocytes were then randomly assigned to treatment with the beta AR agonist isoproterenol (25 nM), the phosphodiesterase inhibitor amrinone (50 microM), or a combination of these compounds and contractile function measurements repeated. Baseline myocyte contractile function was reduced by 32% after HHCA. Isoproternol alone increased myocyte contractile function more than 100% under both normothermic conditions and after HHCA, whereas amrinone alone significantly (60%) improved myocyte contractile function only after HHCA. Amrinone preincubation followed by isoproterenol improved contractile function after HHCA to a greater extent than all other treatment protocols. In contrast, combination treatment under normothermic conditions did not augment myocyte contractile function relative to isoproterenol alone. These findings suggest that amrinone has differential effects on contractile processes. Moreover, the marked improvement of contractile function after HHCA with PDEI pretreatment followed by beta AR stimulation may have implications in treatment strategies for improving myocardial function after cardiopulmonary bypass and provide insight into contractile dysfunction after HHCA. PMID- 7486081 TI - The cost efficacy of hypothetically eliminating adverse anesthetic outcomes from high-risk, but neither low- nor moderate-risk, surgical operations. AB - We tested whether hypothetical improvements in the quality of perioperative care can decrease hospital costs for elective surgical operations. Our case series was constructed retrospectively of consecutive patients who underwent one of the following elective operations at our tertiary care center: lung lobectomy (137), coronary artery bypass (339), kidney transplant (251), total hip replacement (305), laparoscopic cholecystectomy (203), or cesarean section (649). Only hospital costs were included in our study. Eliminating adverse anesthetic outcomes (identified by chart review) entirely would decrease total hospital costs (i.e., for all patients in that group) by less than 0.5% (95% confidence bound < 1.2%) for kidney transplant, 1.8% (< 3.5%) for total hip replacement, 0% for laparoscopic cholecystectomy, and 2.2% (< 6.2%) for cesarean section. In contrast, a hypothetical quality improvement that would have the effect of decreasing costs for the 10% most expensive patients just to the group median would decrease total costs by 30% (15%-44%) for lung lobectomy and 16% (10%-21%) for coronary artery bypass. We conclude that for low- and moderate-risk procedures, hypothetical improvements in the quality of anesthetic care will not reduce costs. However, improving the quality of perioperative care may be cost efficient for high-risk operations. PMID- 7486080 TI - Procaine is effective for minimizing postischemic ventricular fibrillation in cardiac surgery. AB - Procaine hydrochloride was added to cardioplegia and studied for its efficacy in stabilizing the postischemic rhythm after aortic declamping in cardiac surgery. Fifty-six patients scheduled for coronary artery bypass grafting (CABG), were included in a randomized, double-blind study. The patients were anesthetized with isoflurane, low-dose fentanyl, diazepam, and pancuronium. In the study group (28 patients), St. Thomas' Hospital cardioplegic solution II (Plegisol) was prepared with 1 mM procaine. The control group (28 patients) was given the same cardioplegia with saline. Ventricular fibrillation (VF) occurring after declamping was treated with direct current (DC) shock (8-12-12-20 J). There were no significant differences with regard to demographic properties or anesthesiologic and surgical treatment. Two patients (7%) in the procaine group required DC shock for VF, compared to 28 (100%) in the control group (P < 0.001). The amount of lidocaine (mean +/- SEM) given for resistant dysrhythmias was 3.6 mg +/- 3.6 in the procaine group compared to 35.7 mg +/- 9.2 in the control group (P < 0.002). One patient in each group required temporary pacing. The number of synchronized DC shocks for conversion of atrial fibrillation was lower in the procaine group (P < 0.05). The enzyme release the first day after surgery was lower in the procaine group (P < 0.05). Procaine (1 mM) in cardioplegia stabilizes the postischemic rhythm in CABG surgery in humans without any observed adverse effects. PMID- 7486082 TI - Effect of the inhibitor of nitric oxide synthase, NG-nitro-L-arginine methyl ester, on cerebral and myocardial blood flows during hypoxia in the awake dog. AB - The increase in cerebral blood flow (CBF) elicited by moderate hypoxia in anesthetized animals is little attenuated by nitric oxide (NO) synthase inhibitors. However, in previous studies, the effects of NO synthase inhibitors may have been altered by anesthetics. Consequently, we studied the effects of the NO synthase inhibitor, NG-nitro-L-arginine methyl ester (L-NAME), on cerebral and myocardial blood flows during hypoxia in the awake dog. Regional CBF and myocardial blood flows (MBF) were measured under normoxia and hypoxia in 16 beagle dogs after an intravenous (IV) injection of either saline (control, n = 8) or L-NAME 20 mg/kg (n = 8). One week after thoracotomy for catheter insertion, awake dogs were studied during three periods: normoxia and after 2 and 4 h of normocapanic hypoxia in an environmental chamber (FIO2 = 0.10, FICO2 = 0.035, balance N2). At each stage, a bolus injection of L-NAME or saline was followed 15 min later by left atrial injection of radiolabeled microspheres (141Ce, 103Ru, 46Sc) for regional CBF and MBF. After the dogs were killed, the brain and the heart were fixed in 10% formaldehyde, dissected by region and weighed, and radioactivity was measured in a gamma counter. During hypoxia, Pao2 was approximately 45 mm Hg with normal Paco2. In the control group, CBF increased by 45% after 2 h and 48% after 4 h of hypoxia; MBF increased by 69% and 60%, respectively. L-NAME prevented the CBF increase during hypoxia and the MBF increase after 2 h of hypoxia; after 4 h of hypoxia the measurement of MBF was confounded by cardiac dysfunction. These results suggest that NO plays a role in cerebral vasodilation during hypoxia in the awake animal. PMID- 7486084 TI - A prospective study of rectal methohexital: efficacy and side effects in 648 cases. AB - Rectal methohexital has been used for nearly 30 yr in pediatric anesthesia. Despite this long and increasingly varied use, no large prospective series has been published detailing safety and efficacy. This study prospectively evaluated the efficacy, safety, and side effects of this medication in a series of 648 cases. On 553 of 648 occasions (85%), the child fell asleep after a single 30 mg/kg dose of 10% methohexital. Sleep was less likely in patients with myelomeningocele or who were receiving oral phenobarbital or phenytoin. When sleep occurred, the average time to onset of sleep was 6 min. Most patients who remained awake 15 min after drug were sedated. Defecation (10%) and hiccups (13%) were common but benign side effects. Partial airway obstruction and/or desaturation to Spo2 < or = 93% occurred in 26 patients (4%), but was resolved with blow-by oxygen and/or jaw-thrust in all but two cases. These two patients (0.3% of total) required aggressive airway intervention by the supervising anesthesiologist. Apnea did not occur in any patient. Methohexital has a high efficacy rate for sleep (85%) or sedation (96%), and has a relatively rapid onset. Significant respiratory side effects occur infrequently, but can be life threatening if not properly managed. PMID- 7486083 TI - Sodium nitroprusside metabolism in children during hypothermic cardiopulmonary bypass. AB - Ten children, aged 1-7 yr, presenting for repair of complex congenital heart lesions, were prospectively studied. A ketamine, halothane/isoflurane, and fentanyl anesthetic was used. After initiation of hypothermic cardiopulmonary bypass, sodium nitroprusside (SNP) was titrated as necessary to maintain a target mean arterial blood pressure of 35-60 mm Hg. Blood samples drawn immediately prior to starting SNP infusion, every 15 min during infusion, and at 1, 4, and 24 h postinfusion were analyzed for whole blood cyanide (CN-) and serum thiocyanate (SCN-). Blood gas analysis was performed every 30 min during SNP infusion. A maximum CN- level > or = 1.0 micrograms/mL was observed in two children; four others had maximum CN- levels between 0.5 micrograms/mL and 1.0 micrograms/mL (normal, < 0.2 micrograms/mL). No child had a clinically important increase of SCN- subsequent to SNP infusion. There was substantial variability in observed CN accumulation during SNP infusion. CN- levels during the first 60 min correlated with the average SNP rate of administration (P = 0.02). Cyanide levels rapidly decreased after termination of SNP infusion and were undetectable 4 h postinfusion. Despite the short-term increase of CN- level, no child showed biochemical signs of toxicity (acidosis or increased mixed venous oxygen tension). PMID- 7486085 TI - Effect of hemidiaphragmatic paresis caused by interscalene brachial plexus block on breathing pattern, chest wall mechanics, and arterial blood gases. AB - We investigated the effects of hemidiaphragmatic paresis caused by interscalene brachial plexus block on breathing patterns, chest wall mechanics, and arterial blood gas tensions using respiratory inductive plethysmography. Ten healthy patients received interscalene block with 20-40 mL 1.5% lidocaine with epinephrine. Rib cage contribution to tidal volume (%RC) increased from 28.9% +/- 9.7% to 50.0% +/- 8.3% (P < 0.01), respiratory frequency (f) increased from 14.6 +/- 3.2/min to 16.3 +/- 2.4/min (P < 0.05), and PaO2 decreased from 84.7 +/- 7.3 mm Hg to 78.0 +/- 9.5 mm Hg (P < 0.05). No significant changes were observed in tidal volume (VT), minute volume (VE), or PaCO2. These results indicated that VT, VE, and PaCO2 were maintained after interscalene block, apparently by increases in f and %RC to compensate for hemidiaphragmatic paresis caused by interscalene block. Nevertheless, PaO2 was reduced, presumably due to increased ventilation perfusion mismatching. Recognizing that we studied healthy patients, the decrease in PaO2 may be more in patients with cardiopulmonary disease. PMID- 7486086 TI - Intraperitoneal application of bupivacaine during laparoscopic cholecystectomy- risk or benefit? AB - We investigated, in a double-blind study, the effects of intraperitoneal local anesthetics during laparoscopic cholecystectomy. In Part A of the study 30 patients received 50 mL saline 0.9% (A 0), bupivacaine 0.125% (A 125), or bupivacaine 0.25% (A 25) intraperitoneally at the end of surgery. Mean maximum plasma concentrations of bupivacaine reached 0.48 mg/L (range 0.15-0.90 mg/L) in Group A 125 and 1.0 mg/L (0.35-2.10 mg/L) in Group A 25 within 15 min (range, 5 30 min). There was no significant difference in pain scores or opioid consumption (patient-controlled analgesia with piritramid): 24, 28, and 13 mg/24 h among the study groups, respectively (not significant). Postoperative respiratory function deteriorated in comparison to preoperative values in all study groups, but the forced vital capacity was significantly more impaired in Group A .25. In Part B, 24 patients received placebo (B 0) or bupivacaine 0.25% (B 25). Postoperative hypoxemic periods (oxygen saturation < 92%) were significantly more frequent in Group B 25. Considering the questionable benefits and the potential risks, we would not recommend the application of intraperitoneal bupivacaine during laparoscopic cholecystectomy. PMID- 7486087 TI - Analgesia after thoracotomy: effects of epidural fentanyl concentration/infusion rate. AB - After thoracotomy some patients have discomfort, primarily in the rostral portion of their incisions. In this prospective, randomized study in 66 patients after lateral thoracotomy we evaluated whether, for equal fentanyl dosage in micrograms per kilogram, epidural infusion (lumbar catheter) of fentanyl 5 micrograms/mL provided better segmental analgesia (including the rostral portion of the incision) than a 10-micrograms/mL concentration infused at a rate half that used in the 5-micrograms/mL group. Ketorolac was used as an analgesic adjunct for nonincisional pain. Postoperative epidural fentanyl infusion included a 1 microgram/kg initial dose and an initial infusion rate of 1 microgram.kg-1.h-1 in both the 5-micrograms/mL and 10-micrograms/mL groups. Patients were evaluated for comfort level and pain relief while resting, taking a deep breath, coughing, and ambulating at eight times over 3 days using two visual analog scales for overall comfort and a verbal rating score (VRS) for segmental analgesia. There were no significant differences in demographics, surgical procedure, intraoperative fentanyl dose, side effects, rates of epidural fentanyl infusion, or total epidural fentanyl doses at 12, 24, 36, 48, and 60 h postbolus. Analgesia was effective in both groups. Although overall comfort levels were lower (i.e., indicated greater comfort) in the 5-micrograms/mL group in 6 of 8 visual analog scores (VASs) for comfort level and 20 of 24 VRSs for comfort level scores, and mean VRSs for the rostral portion of the incision were lower (i.e., indicated greater comfort) in the 5-micrograms/mL group at 21 of 24 evaluation subsets (one statistically significant), statistical significance was achieved in only six evaluation subsets.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7486088 TI - Epidural ropivacaine infusion for postoperative analgesia after major lower abdominal surgery--a dose finding study. AB - Ropivacaine exhibits less cardiotoxicity and causes less motor block than bupivacaine when used in equianalgesic doses. This makes ropivacaine potentially well suited for epidural infusion for postoperative analgesia. The aim of this study was to determine which of three concentrations of epidurally administered ropivacaine infused for postoperative analgesia would attenuate intravenous opioid analgesia requirements while also minimizing motor block. Forty ASA I-III patients, having major lower abdominal surgery, completed the study. They were randomly assigned to one of four treatment groups: Group S, control, epidural saline (n = 10); Group 1, epidural 0.1% ropivacaine (n = 10); Group 2, epidural 0.2% ropivacaine (n = 10); and Group 3, epidural 0.3% ropivacaine (n = 10). The study was double-blind. Initial epidural analgesia was established with 0.5% ropivacaine, and then general anesthesia induced for surgery. Once in the recovery room, epidural infusions were commenced at 10 mL/h and maintained at that rate for 21 h. Intravenous patient controlled analgesia (PCA) morphine was used as required by the patients for supplemental analgesia. Total PCA morphine use was more over the 21-h period in Group S than all the ropivacaine groups, being significantly so for Group 2 (median values: Group S, 43.3 mg; Group 1, 18.7 mg; Group 2, 7.5 mg; Group 3, 19 mg; for Group 2, P = 0.03). Visual analog scale (VAS) scores on coughing were significantly lower (i.e., less pain) than control for all ropivacaine groups after 4 h of infusion and also for Groups 2 and 3 after 8 h of infusion. (Median VAS (mm) on coughing at 8 h: Group S = 70, Group 1 = 56, Group 2 = 32, Group 3 = 0; for Groups 2 and 3, P < 0.05 compared to Group S). There was a dose-related increase in the amount of motor block, with Group 3 having significantly more motor block than all other groups at 4 and 8 h.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7486089 TI - Efficacy of simulated epidural test doses in adult patients anesthetized with isoflurane: a dose-response study. AB - A recent study demonstrated that an epidural test dose containing 15 micrograms epinephrine was an imperfect marker for intravascular injection during isoflurane anesthesia based on the conventional heart rate (HR) criterion (positive if > or = 20 bpm increase). We have determined the effects of epinephrine doses and isoflurane concentrations on these efficacies in healthy adult patients during isoflurane anesthesia. Eighty patients were randomly assigned to one of four groups according to the simulated test dose injected intravenously (IV) under 1% end-tidal isoflurane and nitrous oxide after endotracheal intubation. The saline group (n = 20) received 3 mL normal saline; the epinephrine 7.5 group (n = 20) received 3 mL 1.5% lidocaine containing 7.5 micrograms epinephrine; the epinephrine 15 and epinephrine 22.5 groups (n = 20 each) received an identical dose and volume of lidocaine but containing 15 and 22.5 micrograms epinephrine, respectively. HR and systolic blood pressure (SBP) were monitored invasively for 4 min after IV injection of the study drug. Although none in the saline group developed a HR increase > or = 20 bpm, 2, 14, and 12 patients elicited positive responses in the epinephrine 7.5, 15, and 22.5 groups (10%, 70%, and 60% sensitivities), respectively. If a positive HR response was defined by an increase of 10 bpm, sensitivities were 55%, 100%, and 100% in the epinephrine 7.5, 15, and 22.5 groups, respectively. On the other hand, none in the saline group, 12 in the epinephrine 7.5 group, and all patients in the epinephrine 15 and 22.5 groups developed maximum SBP increases > or = 15 mm Hg.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7486090 TI - Pneumothorax during laparoscopic fundoplication: diagnosis and treatment with positive end-expiratory pressure. AB - Pneumothorax can develop during laparoscopy, particularly during laparoscopic fundoplication, since the left parietal pleura is exposed and can be torn during dissection in the diaphragmatic hiatus. Such an event will result in specific pathophysiologic changes, since CO2, under pressure in the abdominal cavity, will pass into the pleural space. The aim of this study was to document the pathophysiologic changes induced by pneumothorax, and to evaluate the benefit of positive end-expiratory pressure (PEEP) to treat pneumothorax. Forty-six ASA physical status I and II patients scheduled for laparoscopic fundoplication were monitored extensively; heart rate, mean arterial pressure, end-tidal CO2 (PETCO2), oxygen saturation of hemoglobin (Spo2), minute ventilation, tidal volume, dynamic total lung thorax compliance, and airway pressures were recorded. In 25 patients, oxygen uptake, CO2 elimination and arterial blood gases were also measured. Pneumothorax was diagnosed in seven patients. It resulted in the following pathophysiologic changes: decrease in total lung thorax compliance, increase in airway pressures, and increase in CO2 absorption. Consequently, PACO2 and PETCO2 also increased. Spo2, however, remained normal. The use of PEEP largely corrected these respiratory changes. None of these pneumothoraces required drainage. These data suggest that pneumothorax is common during laparoscopic fundoplication. Early diagnosis is possible by simultaneous monitoring of PETCO2, total lung thorax compliance, and airway pressures. Finally, treatment with PEEP provides an alternative to chest tube placement when pneumothorax is secondary to passage of peritoneal CO2 into the interpleural space. PMID- 7486091 TI - Adhesion molecules and inflammation: the next targets for perioperative organ protection? PMID- 7486092 TI - Hyperbaric lidocaine spinal anesthesia: do we need an alternative? PMID- 7486093 TI - Circulating adhesion molecules in pediatric cardiac surgery. AB - Cardiopulmonary bypass (CPB) may be associated with the risk of a "whole body inflammation." Adhesion molecules, such as endothelial leukocyte adhesion molecule (ELAM-1), intercellular adhesion molecule-1 (ICAM-1), and vascular cell adhesion molecule-1 (VCAM-1), seem to play a pivotal role in the inflammatory response. Soluble forms of these adhesion molecules may serve as markers of endothelial activation or damage. To elucidate whether plasma levels of soluble adhesion molecules differ between pediatric and adult cardiac surgery patients, 15 consecutive children younger than 5 yr undergoing CPB were prospectively studied and compared with adults scheduled for elective coronary artery bypass grafting and valve replacement. Plasma levels of circulating (soluble) adhesion molecules (sELAM-1, sICAM-1, sVCAM-1) were measured from arterial blood samples using enzyme-linked immunosorbent assays after induction of anesthesia (= "baseline"), during CPB, at the end of surgery, and on postoperative days 1 and 2. At baseline, plasma levels of all three soluble adhesion molecules were significantly higher in children than in adults. sELAM-1 and sICAM-1 plasma concentrations were even beyond normal in the children (sELAM-1: 88.8 +/- 13.8 ng/mL; sICAM-1: 349 +/- 27 ng/mL). During CPB and until the end of surgery, plasma levels of all adhesion molecules decreased in the children and remained almost unchanged in the adults. In the children, sELAM-1 remained lower than baseline values until the second postoperative day (45.2 +/- 12.2 ng/mL), whereas sICAM-1 increased in the postbypass period without, however, reaching baseline values (254 +/- 40 ng/mL).(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7486094 TI - Recovery and complications after tonsillectomy in children: a comparison of ketorolac and morphine. AB - Ninety-six children received morphine 0.1 mg/kg (n = 47) or ketorolac 1 mg/kg (n = 49) intravenously (IV) in a prospective, randomized, double-blind fashion, after tonsillectomy. Recovery variables and complications were recorded while subjects were in the hospital and parent(s) were contacted 24 h and 14 days after surgery. There were no differences in demographics, surgical management, awakening time, oxygen requirements, or time to readiness for postanesthesia care unit (PACU) discharge or discharge home between the two groups. Ketorolac subjects had fewer emetic episodes than morphine subjects (median 1 vs 3; P = 0.006) and were less likely to have more than two episodes of emesis after PACU discharge (9/49 vs 22/47; P = 0.007). Ketorolac subjects had more major bleeding (bleeding requiring intervention; 5/49 vs 0/47, one-tailed P = 0.03) and more bleeding episodes (0.22 episodes/subject vs 0.04 episodes/subject, P < 0.05) in the first 24 h after surgery, but no greater overall incidence of bleeding than the morphine subjects. In children having tonsillectomy, ketorolac, compared to morphine, reduced the number of emetic episodes after PACU discharge, but did not hasten awakening, readiness for PACU discharge or discharge home, and increased the likelihood of major bleeding in the first 24 h after surgery. PMID- 7486095 TI - Intraoperative blood loss in pediatric liver transplantation: analysis of preoperative risk factors. AB - The relative contribution of 14 preoperative risk factors to a high intraoperative blood loss was studied in 95 consecutive first pediatric orthotopic liver transplantations (OLT). Patients were distributed in two groups according to red blood cell (RBC) requirements. Wide interindividual RBC requirements were observed (median, 79 mL/kg; range, 4-586). The upper quartile of the population was defined as the high blood loss group and required 123 mL/kg or more (median, 161). On univariate analysis, the high blood loss group had a significantly higher proportion of patients with portal vein hypoplasia, intraabdominal malformations, signs of severe liver failure (encephalopathy, ascites, prolonged prothrombin time), and requiring inpatient support. Age, previous abdominal surgery, and platelet count had no prognostic value. All variables used in the univariate analysis were included in a stepwise logistic regression analysis. Only presence of portal vein hypoplasia, inpatient support, and use of a reduced-size liver graft were independently associated with a high blood loss. Adjusted odds ratios were 40.4 (95% confidence interval; 5.9-278), 5.4 (1.6-17.9), and 3.8 (0.9-15.2), respectively, highlighting the importance of portal vein hypoplasia as a risk factor for high blood loss. PMID- 7486097 TI - Epidural clonidine or sufentanil for intraoperative and postoperative analgesia. AB - This study contrasts the efficacy and side effects of epidural clonidine and sufentanil in the perioperative period. Using a randomized, prospective, double blind study design, 40 patients undergoing abdominal surgery under propofol/nitrous oxide anesthesia were enrolled. Before anesthesia, an epidural catheter was inserted at the L1-L2 interspace. At induction of anesthesia, the patients received epidurally either clonidine (4 micrograms/kg in 10 mL) infused in 20 min followed by a 2-micrograms.kg-1.h-1 infusion (5 mL/h) during 12 h (Group 1) or sufentanil (0.5 microgram/kg in 10 mL) in 20 min followed by a 0.25 microgram.kg-1.h-1 infusion (5 mL/h) during 12 h (Group 2). Intraoperatively, increases in arterial blood pressure and heart rate not responding to propofol bolus (0.5 mg/kg) were treated with a bolus of intravenous (IV) sufentanil 0.035 microgram/kg. Postoperatively, IV sufentanil boluses (5 micrograms) were given through a patient-controlled analgesia (PCA) device. Postoperative analgesia was assessed by recording the IV PCA sufentanil requirements and the patients' visual analog scale (VAS) at 3, 6, 12, 18, 24, 36, and 48 h. Sedation analog scales and side effects were also recorded. Plasma clonidine and sufentanil concentrations were measured after 20 min and 6, 12, and 24 h. The number of reinjections of propofol (n = 1.6 +/- 1.6 in Group 1 vs 6.5 +/- 4.0 in Group 2) and of IV sufentanil (n = 0.6 +/- 0.8 in Group 1 vs 3.8 +/- 3.7 in Group 2) was significantly reduced (P < 0.001) in the epidural clonidine group. In the early postoperative period, pain scores and rescue analgesic requirements were very low in both groups.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7486096 TI - Transient neurologic symptoms after spinal anesthesia. AB - We recently reported several cases consistent with transient radicular irritation after spinal anesthesia with hyperbaric 5% lidocaine. The present prospective, blind, nonrandomized study was performed to determine the incidence of these transient neurologic symptoms and to identify factors that might be associated with their occurrence. We studied 270 patients scheduled for gynecologic or obstetric procedures under spinal anesthesia. For spinal anesthesia, either 5% lidocaine in 7.5% glucose or 0.5% bupivacaine in 8.5% glucose was used. Patients were evaluated on postoperative day 3 by a quality assurance nurse who was unaware of the drug given or details of the anesthetic technique. Transient neurologic symptoms were observed in 37% of patients receiving 5% lidocaine, whereas only one patient receiving 0.5% bupivacaine had transient hypesthesia of the lateral aspect of the right foot. These results suggest that symptoms were the result of a specific drug effect. However, because of the limitations of the study one cannot conclude that lidocaine per se was the cause. PMID- 7486098 TI - Pharmacokinetics of ropivacaine and bupivacaine during 21 hours of continuous epidural infusion in healthy male volunteers. AB - The aim of the present study was to evaluate the pharmacokinetics of ropivacaine and to compare the results with those of bupivacaine during prolonged epidural infusion. Ropivacaine 1, 2, or 3 mg/mL (0.1%, 0.2%, or 0.3%), bupivacaine 2.5 mg/mL (0.25%), or placebo (sodium chloride 0.9%) was given randomly and in a double-blind manner to five parallel treatment groups (37 healthy volunteers) as a continuous epidural infusion for 21 h. A 10-mL epidural bolus dose was first given, and the epidural infusion was started immediately afterward. The subjects received 10 mL/h corresponding to infusion rates of 10, 20, or 30 mg/h ropivacaine and 25 mg/h bupivacaine, respectively. Peripheral blood samples for measurements of ropivacaine or bupivacaine were taken during a 25-h period. The total plasma concentration increased continuously but seemed to reach a plateau (C5-10h) after approximately 5 h infusion, remaining fairly constant up to approximately 10 h after the start of administration. The C5-10h values were proportional to the dose of ropivacaine and were estimated as 0.3, 0.6, and 0.9 mg/L, and for bupivacaine as 0.7 mg/L. During the subsequent infusion the plasma concentration increased, with maximum plasma levels at the end of the infusion and with corresponding values of 0.4, 0.9, 1.2, and 0.9 mg/L. The highest individual plasma concentration was 1.7 mg/L (20 mg/h), and no patient showed signs of toxic systemic plasma levels. The free concentrations also increased continuously during the infusion. The free fraction was independent of the dose (6.1% for ropivacaine and 4.8% for bupivacaine).(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7486099 TI - The comparison of epidural fentanyl, epidural lidocaine, and intravenous fentanyl in patients undergoing gastrectomy. AB - This study was conducted prospectively to compare the effect of epidural fentanyl (EP-F), epidural lidocaine (EP-L), and intravenous fentanyl (IV-F) on hemodynamic and hormonal responses to surgery and postoperative analgesic requirement in 30 patients undergoing gastrectomy during isoflurane anesthesia. An epidural catheter was placed via the T8-9 interspace. Group EP-F received fentanyl 2 micrograms/kg in 10 mL saline, and Group EP-L, 10 mL 1.5% lidocaine, epidurally; Group IV-F was given fentanyl, 2 micrograms/kg, IV. Fifty percent of the original dose was repeated every hour. Hemodynamic data and plasma hormonal levels were compared between those before and those at 1 h after skin incision. The total number of analgesic administrations within the first 48 h postoperatively were compared. Group EP-L developed more frequent episodes of hypotension. Group IV-F required higher isoflurane concentrations and the plasma epinephrine levels increased more than in Groups EP-F and EP-L. In Groups EP-L and IV-F, the plasma antidiuretic hormone (ADH) level increased more than in Group EP-F. In Groups EP F and IV-F, the plasma cortisol and adrenocorticotropic hormone (ACTH) levels increased more than in Group EP-L. The use of postoperative analgesics was significantly less in Group EP-F. In conclusion, in Group EP-F, attenuated hormonal responses to surgery was accompanied with less hypotension and postoperative analgesic requirements were reduced. PMID- 7486100 TI - Continuous intravenous administration of ketorolac reduces pain and morphine consumption after total hip or knee arthroplasty. AB - The purpose of this study was to determine the analgesic efficacy, opioid-sparing effect, and tolerability of ketorolac administered as an intravenous (i.v.) bolus followed by a continuous infusion after total hip or knee arthroplasty. After general anesthesia, patients received either placebo or ketorolac 30 mg i.v. as a bolus over 15-30 s followed by a continuous i.v. infusion of ketorolac 5 mg/h for 24 h. All patients received patient-controlled i.v. morphine with no background infusion. Patients were assessed at 2, 4, 6, and 24 postoperatively with respect to analgesia, morphine consumption, side effects, and blood loss. Patients receiving ketorolac reported were less sedated and required fewer antiemetics. There was no difference in blood loss. Patients receiving ketorolac reported better analgesia and used less morphine (35% for hips and 44% for knees) than those receiving placebo. PMID- 7486101 TI - Cellular localization of dopamine D2 receptor messenger RNA in the rat trigeminal ganglion. AB - The actions of dopamine are mediated by specific, high-affinity, G protein coupled receptors. Multiple subtypes of dopamine receptors have been characterized, including the D2 subtype (D2R). Cells within the dorsal root and petrosal ganglia of the rat express D2R messenger RNA (mRNA) consistent with D2R expression by primary sensory neurons. We hypothesized that neurons of the trigeminal ganglion express D2R mRNA. Total cellular RNA from rat trigeminal ganglia was analyzed on Northern blots under high stringency conditions. Hybridization of trigeminal ganglion RNA resulted in a signal which comigrated with striatal, pituitary, and hypothalamic D2R mRNA. To determine the distribution of D2R expressing cells in the trigeminal ganglion, cryostat sections were analyzed by in situ hybridization followed by emulsion autoradiography. We identified a population of clustered cells labeled with dense grain concentrations over their cytoplasms. These findings demonstrate the expression of D2 dopamine receptor mRNA in discrete subpopulations of neurons in the rat trigeminal ganglion. Our observations suggest that drugs active at dopamine receptors of the D2 subtype are potential modulators of sensory activity of neurons whose cell bodies reside in the trigeminal ganglion. D2 dopamine receptors may thus have a role in clinical pain syndromes involving the head and neck. PMID- 7486103 TI - Propofol-associated dilation of rat distal coronary arteries is mediated by multiple substances, including endothelium-derived nitric oxide. AB - Previous in vitro studies on the effect of propofol on coronary arteries have shown variable results, ranging from constriction to no effect to dilation. Although most of these studies reported that the observed effect is endothelium independent, propofol also releases nitric oxide from cultured porcine endothelial cells. The present study examines the direct effect of propofol in rat distal coronary arteries in vitro, especially in regard to endothelial dependence and involvement of the adenosine triphosphate (ATP)-sensitive potassium channels (KATP channels). Forty-three subepicardial arteries (size 91.1 +/- 15.8 microns) from Wistar rats were studied in vitro in a no-flow, pressurized (40 mm Hg) state, using an optical density video detection system. After preconstriction with the thromboxane analog U46619 1 microM, relaxation responses to increasing concentrations of propofol (10(-6)-10(-4) M) were measured after 1) endothelial denudation, 2) pretreatment with the nitric oxide synthase inhibitor NG-nitro-L-arginine (L-NNA), 3) pretreatment with the cyclooxygenase inhibitor indomethacin, 4) pretreatment with the KATP channel blocker glibenclamide, or 5) no intervention (control). Propofol produced a significant concentration-dependent vasodilation of the U46619-preconstricted coronary arteries. This effect was significantly attenuated by endothelial denudation, pretreatment with L-NNA, or indomethacin, but was not affected by glibenclamide. We conclude that propofol has a direct vasodilatory effect on distal coronary arteries in rats. This effect is primarily endothelium-dependent and is mediated by multiple substances, including nitric oxide (NO) and a vasodilatory prostanoid. The effect is not mediated by opening of the K(ATP) channels. PMID- 7486102 TI - Sevoflurane versus desflurane for outpatient anesthesia: a comparison of maintenance and recovery profiles. AB - The recovery characteristics of desflurane and sevoflurane were compared when used for maintenance of ambulatory anesthesia. After obtaining informed consent, 42 healthy, unpremedicated women undergoing laparoscopic sterilization procedures were studied. Anesthesia was induced with propofol, 1.5-2.0 mg/kg, and maintained with either desflurane 3%-6% (n = 21) or sevoflurane 1%-2% (n = 21) with 60% nitrous oxide in oxygen. Intraoperative analgesia and neuromuscular block was achieved using fentanyl and vecuronium, respectively. The inhaled anesthetics were titrated to achieve an adequate clinical "depth of anesthesia" and to maintain mean arterial pressure (MAP) within 20% of the preinduction baseline values. Visual analog scales (VAS) and the digit-symbol substitution test (DSST) were performed preoperatively and at 30-min intervals during the recovery period. There were no differences between the two groups in the total doses of propofol, fentanyl, or vecuronium. Heart rate (HR) values were lower in the sevoflurane group during the induction-to-incision period. However, HR and MAP were otherwise similar during the maintenance and recovery periods. Use of desflurane led to a more rapid emergence (4.8 +/- 2.4 vs 7.8 +/- 3.8 min) and shorter time to extubation (5.1 +/- 2.2 vs 8.2 +/- 4.2 min) compared to sevoflurane (mean values +/- SD). Intermediate recovery times, postoperative VAS and DSST scores, and side effects were similar in the two treatment groups. Although sevoflurane was associated with a slower emergence from anesthesia than desflurane after laparoscopic surgery, recovery of cognitive function and discharge times were similar in the two anesthetic groups. Thus, it would appear that sevoflurane is an acceptable alternative to desflurane for maintenance of outpatient anesthesia.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7486104 TI - A comparison of four bedside methods of hemoglobin assessment during cardiac surgery. AB - The purpose of this study was to compare the accuracy of conductivity, adjusted conductivity, photometric, and centrifugation methods of measuring or estimating hemoglobin (Hb) with Coulter measured HB as the reference. These bedside methods were studied in 25 cardiac surgery patients during euvolemia and hemodilution and after salvaged autologous red blood cell transfusion. In vivo patient blood samples were obtained before induction, at the start of cardiopulmonary bypass (CPB), after CPB, and after blood transfusion. In 10 patients, blood was sampled in vitro from units of processed blood. Hb values were determined using conductivity by Stat-Crit, adjusted conductivity by Nova Stat Profile 9, bedside photometry by HemoCue, and centrifugation methods. The calculated bias values of Coulter test method Hb (mean +/- SD) for in vivo patient blood samples (n = 90) were: Stat-Crit = 0.6 +/- 0.8 g/dL; Nova Stat Profile 9 = -0.7 +/- 0.4 g/dL; HemoCue = -0.1 +/- 0.2 g/dL; and centrifuge = 0.1 +/- 0.5 g/dL (P < 0.0001). Hb bias values (g/dL) for in vitro samples (n = 10) obtained from processed blood were Stat-Crit = 5.1 +/- 0.6; Nova Stat Profile 9 = 3.0 +2- 0.6; HemoCue = 0.4 +/ 0.4; and centrifuge = 0.6 +/- 0.3 (P < 0.0001). Hb assessment by different test methods may be significantly affected during hemodilution and after blood transfusion. In vitro conditions exaggerated the inaccuracy of conductivity and adjusted conductivity Hb estimates. The rank order of closest approximation to the Coulter measurement for all in vivo blood samples was provided by bedside photometry, followed by centrifugation, adjusted conductivity, and uncorrected conductivity methods. PMID- 7486105 TI - Transcranial Doppler sonography indicates critical brain perfusion during hemorrhagic hypotension in dogs. AB - This study investigated the effects of hemorrhagic hypotension on cerebral blood flow velocity and brain electrical activity (by electroencephalogram [EEG]). Eleven mongrel dogs were anesthetized with isoflurane (1 minimum alveolar anesthetic concentration [MAC]) and catheters were placed into both femoral arteries and veins for mean arterial blood pressure (MAP) measurement, blood withdrawal, and drug administration. Brain temperature, arterial blood gases, and pH were maintained constant. EEG was recorded from temporoparietal recording sites versus a frontal reference. A pulsed transcranial Doppler (TCD) probe (2 MHz, Transpect, Medasonics) was placed on the dura via a temporal bone window to measure mean (Vmean, cm/s) and diastolic blood flow velocity (Vdiast, cm/s) in the middle cerebral artery. At the end of the surgical preparation, isoflurane was discontinued and all animals received fentanyl (bolus, 25 micrograms/kg intravenously (IV); infusion, 50 micrograms.kg-1.h-1 IV) plus 50% N2O/O2 during 30 min of equilibration. After recordings of baseline data, the dogs were hemorrhaged at a rate of 80-100 mL/min. The observation interval was 14 min. EEG spectral edge frequency (SEF 95%) and Vmean did not change when MAP was decreased from 109 +/- 10 to 63 +/- 7 mm Hg. This indicates preserved neuronal function and intact autoregulation of cerebral blood flow. Below MAP of 49 +/- 9 mm Hg, a shift of the EEG to lower frequencies was associated with decreases in Vmean and Vdiast. EEG burst suppression occurred at a MAP of 31 +/- 7 mm Hg, paralleled by a loss of the diastolic flow velocity pattern.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7486106 TI - Mechanisms of dexmedetomidine-induced cerebrovascular effects in canine in vivo experiments. AB - Dexmedetomidine decreases cerebral blood flow without significantly affecting cerebral oxygen consumption in anesthetized dogs. To assess the direct cerebrovascular effects of dexmedetomidine, we investigated the responses of vasomotor tone to topical application of dexmedetomidine to pial vessels in vivo, using a parietal cranial window. Forty-one dogs were anesthetized with pentobarbital. In 20 dogs, we topically applied six concentrations of dexmedetomidine solution (10(-8), 10(-7), 10(-6), 10(-5), 10(-4), 10(-3) M) and directly measured pial arterial and venous diameters. In 10 dogs, the inhibitory effects of pretreatment of pial vessels with 10(-5) M yohimbine were examined after the application of 10(-5) dexmedetomidine. In the remaining 11 dogs, the effects of 10(-3) M dexmedetomidine were evaluated in the presence of N omega nitro-L-arginine methyl ester (L-NAME), glibenclamide, or propranolol. Dexmedetomidine significantly constricted pial arteries and veins in a concentration-dependent manner (10(-7) M to 10(-4) M; P < 0.05). Yohimbine blocked dexmedetomidine-induced constriction of pial vessels (both large and small arteries and large veins P < 0.0001; small veins P < 0.005). However, when the highest concentration of dexmedetomidine (10(-3) M) was administered under the window, pial vessel diameter was not significantly altered. In the presence of glibenclamide, 10(-7) and 10(-3) M dexmedetomidine induced a significant decrease in pial arterial diameter compared with 10(-7) and 10(-3) M dexmedetomidine solution alone, respectively (P < 0.05). L-NAME or propranolol did not affect the dexmedetomidine-induced constriction.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7486107 TI - Biochemical characterization of the stimulatory effects of halothane and propofol on purified brain protein kinase C. AB - Halothane and propofol stimulate activation of protein kinase C (PKC) in the presence of physiologically relevant lipid bilayer vesicles in vitro. The mechanism of this stimulation was characterized by analyzing the effects of halothane and propofol on the activation of purified rat brain PKC by its three essential activators, phosphatidylserine, diacylglycerol, and Ca2+, each of which is known to interact with the regulatory domain. Clinically relevant concentrations of halothane (2.4 vol%) and propofol (200 microM) increased the Vmax without affecting the Km for phosphorylation of the artificial substrate histone H1 by PKC, and increased the sensitivity of PKC to activation by phosphatidylserine, diacylglycerol, and Ca2+. Halothane reduced the EC50 values for phosphatidylserine from 18 +/- 2.5 to 11 +/- 0.6 mol% (P < 0.05), for diacylglycerol from 1.6 +/- 0.3 to 0.87 +/- 0.2 mol% (P < 0.05) and for free Ca2+ from 4.5 +/- 1.0 to 2.8 +/- 0.4 microM (P < 0.05). Propofol reduced the EC50 values for phosphatidylserine from 18 +/- 1.9 to 11 +/- 1.2 mol% (P < 0.01), for diacylglycerol from 2.5 +/- 0.3 to 1.2 +/- 0.4 mol% (P < 0.01) and for free Ca2+ from 2.8 +/- 0.7 to 1.9 +/- 0.2 microM (P < 0.05). The IC50 values for inhibition of PKC activity by the regulatory domain-specific PKC inhibitor sphingosine were increased from 20 +/- 1.5 to 26 +/- 0.6 microM (P < 0.01) by halothane and from 24 +/- 4.8 to 34 +/- 4.8 microM (P < 0.05) by propofol.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7486108 TI - Cortical reactivity during isoflurane burst-suppression anesthesia. AB - We studied cortical reactivity to auditory, visual, and somatosensory stimuli during moderate and deep levels of isoflurane anesthesia at which the electroencephalogram (EEG) showed burst suppression patterns, defined as alternating high amplitude bursts and periods of suppressed background activity. Fifteen patients scheduled for gynecologic surgery were anesthetized with isoflurane until burst suppression appeared in the EEG. During steady state burst suppression at 1.5 end-tidal isoflurane concentration (ETisof), each patient was given a 5-min interval each of episodes of visual, auditory, and somatosensory stimulation. During the 5-min interval of visual stimulation the patient was given 3-s episodes of 60 flashes, 4 ms duration each, at a 20-Hz frequency via redlight-emitting diode goggles. Corresponding auditory and somatosensory stimulation consisted of 60 clicks (80 dB, 0.1 ms, 20 Hz) via earphones and 60 pulses to the median nerve at the wrist (20 mA, 0.2 ms, 20 Hz). The 3-s episodes of stimulation were given at irregular intervals ranging from 5 to 20 s. End tidal isoflurane was then increased by 0.3 vol% and 15 min later the stimulation sequence was repeated. During anesthesia at 1.5 +/- 0.1 ETisof all stimulus modalities readily evoked bursts. One hundred percent of visual stimuli, 98% +/- 4% of somatosensory stimuli, and 94% +/- 9% of auditory stimuli, given during EEG suppression, evoked bursts. Somatosensory and visual stimulation evoked bursts at both onset and offset of the 3-s episodes of stimuli. The responses to auditory stimuli were related mainly to the ending of the 3-s episode of clicks.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7486109 TI - Tolerance to nitroglycerin in vascular smooth muscle cells is not affected by the level of intracellular glutathione or L-cysteine. AB - A major hypothesis for the mechanism of tolerance to nitroglycerin (NTG) is that continued use causes a decrease in thiol donors within the vascular smooth muscle cell that are essential for the effect of NTG. We tested this idea directly in the target cell. NTG tolerance, measured as reduced formation of intracellular cyclic guanosine monophosphate (cGMP), was induced in pig coronary smooth muscle cells. The consequence of altering intracellular levels of the thiol donors, glutathione (GSH) and L-cysteine (L-cys), was determined. Incubating cells with 100 microM NTG for 1 h caused an 83% reduction in cGMP formation in response to acute readministration of 200 microM NTG for 2 min but was not associated with a reduction in intracellular GSH or L-cys. This result was not altered when intracellular GSH levels were increased three-fold by including 1 mM GSH in the incubation buffer. Also, recovery from tolerance was not affected by supplementation with GSH. Further, the response of cGMP to NTG was not altered by inhibiting the synthesis of GSH and lowering intracellular levels of GSH by 77%. Similar findings were made with supplemental L-cys or N-acetyl-L-cysteine. These results do not support the hypothesis that tolerance to NTG is the result of a reduction of the thiol donors GSH and L-cys within vascular smooth muscle cells. PMID- 7486111 TI - Does hypercarbia develop faster during laparoscopic herniorrhaphy than during laparoscopic cholecystectomy? Assessment with continuous blood gas monitoring. AB - The use of CO2 to create and maintain a pneumoperitoneum during laparoscopic surgery may lead to hypercarbia and acidosis. CO2 is also insufflated into the preperitoneal space to create and maintain a pneumopreperitoneum for laparoscopic herniorrhaphy. This study examined the influence of CO2 pneumopreperitoneum on the development of hypercarbia and acidosis assessed with continuous intraarterial blood gas monitoring. Changes in blood gas values were measured with both continuous intraarterial and intermittent blood gas monitoring. Over a 4-mo period, blood gas values of 14 patients undergoing laparoscopic herniorrhaphy (pneumopreperitoneum) were compared with those of 13 patients undergoing laparoscopic cholecystectomy (pneumoperitoneum) in a tertiary referral center. Additionally, heart rate and blood pressure were measured during stable ventilation at constant insufflation pressure. Pneumopreperitoneum resulted in a significantly faster development of hypercarbia (P = 0.023) and acidosis (P = 0.027) than pneumoperitoneum. These results were not explained when corrected for changes in hemodynamic and ventilatory variables using analysis of covariance. We conclude that the more rapid development of hypercarbia and acidosis during pneumopreperitoneum can be explained by increased CO2 absorption through an increasing gas exchange area during the procedure and through a larger wound bed. PMID- 7486110 TI - Effects of 6% hydroxyethyl starch and 3% modified fluid gelatin on intravascular volume and coagulation during intraoperative hemodilution. AB - In the perioperative period, artificial colloids are most often infused in doses of 500-1000 mL intravenously. This randomized study compared the effects on intravascular volume and coagulation of approximately 2000 mL of two isooncotic artificial colloids: 6% hydroxyethyl starch (HES; MW 200,000; substitution ratio 0.40-0.55) and 3% modified fluid gelatin (GEL). We hypothesized more pronounced hypocoagulation with HES and a weaker intravascular volume effect of GEL. Forty two patients, scheduled for primary total hip replacement, were allocated randomly to receive HES or GEL during acute normovolemic hemodilution and subsequent further intraoperative hemodilution. Blood samples were taken before and after 500 mL and 1000 mL of acute normovolemic hemodilution; intraoperatively after 20 mL/kg of artificial colloid and at the end of colloid infusion; on arrival in the recovery room; and 3 h later. We quantified: 1) coagulation variables; 2) blood loss; 3) hemodynamic stability; 4) necessary infusion volume; 5) interstitial extravasation, calculated from plasma volunteers measured using albumin marked with technetium-99m and iodine-125, respectively; 6) percentage volume effect at the end of the study as well as hematocrit, total serum protein, and colloid osmotic pressure. Intraoperative volume therapy was guided by radial systolic pressure and systolic pressure variation, mixed venous hemoglobin saturation in the pulmonary artery, and pulmonary capillary occlusion pressure.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7486112 TI - Beta 2-adrenoceptor density of human lymphocytes after nitroprusside-induced hypotension. AB - The present study was undertaken to assess the influence of nitroprusside-induced hypotension on beta 2-adrenoceptor density. Twenty-four patients undergoing nose septum corrections under general anesthesia were allocated randomly to a nitroprusside or control group. beta 2-Receptor density on lymphocytes was measured by binding studies using (-)125-iodocyanopindolol. Lymphocyte subpopulations B, T, Thelper, Tsuppressor, and natural killer cells were determined simultaneously by flow cytometry. Five of 12 nitroprusside-treated patients developed significant intraoperative increases of epinephrine levels (+69% versus preoperatively) which were not seen in the remaining seven patients. In these five patients, beta 2-receptor density of unfractionated lymphocytes was 26% lower (P < 0.05) on the first day after surgery compared with preoperative values. Since no changes in proportions of lymphocyte subpopulations were observed, these results are not caused by redistribution phenomena inducing a decrease of subsets with a high number of beta receptors. These findings suggest that beta 2-adrenergic responsiveness might be diminished after nitroprusside treatment in some patients. PMID- 7486114 TI - Decreases in anesthesia-controlled time cannot permit one additional surgical operation to be reliably scheduled during the workday. AB - We tested whether anesthesiologists can decrease operating room (OR) costs by working more quickly. Anesthesia-controlled time (ACT) was defined as the sum of 1) the time starting when the patient enters an OR until preparation or surgical positioning can begin plus 2) the time starting when the dressing is finished and ending when the patient leaves the OR. Case time was defined as the time starting when one patient undergoing an operation leaves an OR and ending when the next patient undergoing the same operation leaves the OR. An actual case series was constructed of 709 consecutive patients who underwent one of 11 elective operations at a tertiary care center. Statistical analysis of measured OR times showed that ACt would have to be decreased by more than 100% to permit one additional scheduled, short (30-min) operation to be performed in an OR during an 8-h workday after a prior series of cases, each lasting more than 45 min. Anesthesiologists alone cannot reasonably decrease case times sufficiently to permit one extra case to be reliably scheduled during a workday. Methods to decrease ACT (e.g., using preoperative intravenous catheter teams, procedure rooms, and/or shorter acting drugs) may simply increase costs. PMID- 7486113 TI - Loss of alveolar macrophages during anesthesia and operation in humans. AB - Pulmonary macrophages play an important role in the host defense against infection, and the importance of this role is probably enhanced when the upper airway defenses are circumvented by endotracheal intubation. Studies in animals suggest that exposure to volatile anesthetics compromises the viability and function of alveolar macrophages. We studied the effect of surgery and anesthesia on the alveolar macrophages of 41 human subjects undergoing lower abdominal procedures of varying lengths during nitrous oxide-isoflurane anesthesia. Alveolar macrophages were harvested from bronchoalveolar lavage fluid obtained before incision and compared to those recovered just before emergence from anesthesia. Macrophages were analyzed for aggregation and viability, assessed by the ability of viable cells to exclude trypan blue dye. Operations lasting 2 h or less led to little aggregation and had little effect on viability. However, there was a strong correlation between loss of macrophages and the duration of surgery and anesthesia. Aggregation increased and viability decreased as a function of procedure length. Studies are needed to determine whether prolonged surgery contributes to the incidence of postoperative pulmonary complications by disturbing the function and survival of alveolar macrophages in humans. PMID- 7486115 TI - Propofol blood concentration and the Bispectral Index predict suppression of learning during propofol/epidural anesthesia in volunteers. AB - Propofol is often used for sedation during regional anesthesia. We tested the hypothesis that propofol blood concentration, the Bispectral Index and the 95% spectral edge frequency predict suppression of learning during propofol/epidural anesthesia in volunteers. In addition, we tested the hypothesis that the Bispectral Index is linearly related to propofol blood concentration. Fourteen healthy, male volunteers were studied on three randomly ordered days: no propofol, target propofol blood concentration 1 microgram/mL, and target propofol blood concentration 2 micrograms/mL. Each day, epidural anesthesia (approximately T11 level) was induced using 2% 2-chloroprocaine. Propofol was infused by a computer-controlled pump, and propofol concentration measured in central venous blood. We administered a Trivial Pursuit-type question task on all 3 days. The electroencephalogram was monitored continuously (Fp1, Fp2; reference, Cz; ground, mastoid). Propofol caused concentration-related impairment of learning. The propofol blood concentration suppressing learning by 50% was 0.66 +/- 0.1 microgram/mL. The Bispectral Index value when learning was suppressed by 50% was 91 +/- 1. In contrast, the 95% spectral edge frequency did not correlate well with learning. The Bispectral Index decreased linearly as propofol blood concentration increased (Bispectral Index = -7.4.[propofol] + 90; r2 = 0.47, n = 278). There was no significant correlation between the 95% spectral edge frequency and propofol concentration. In order to suppress learning, propofol blood concentrations reported to produce amnesia may be targeted. Alternatively, the Bispectral Index may be used to predict anesthetic effect during propofol sedation. PMID- 7486116 TI - Performance of computer-controlled infusion of propofol: an evaluation of five pharmacokinetic parameter sets. AB - Computer-controlled infusion of propofol is used with increasing frequency for the induction and maintenance of anesthesia. The performance of computer controlled infusion devices is highly dependent on how well the implemented pharmacokinetic parameter set matches the pharmacokinetics of the patient. This study examined the performance of a computer-controlled infusion device when provided with five different pharmacokinetic parameter sets of propofol in female patients. The infusion rate-time data that had been stored on a disk from 19 female patients who had been given propofol by computer-controlled infusion, using the pharmacokinetic parameter set from Gepts et al. (Anesth Analg 1987;66:1256-63), were entered into a computer simulation program to recalculate predicted propofol concentrations that would have been obtained with four other pharmacokinetic parameter (Shafer et al., Anesthesiology 1988;69:348-56; Kirkpatrick et al., Br J Anesth 1988;60:146-50; Cockshott et al., Br J Anesth 1987;59:941P; Tackley et al., Br J Anesth, 1989;62:46-53) sets of propofol, had these been implemented. The performance error (PE) was determined for each measured blood propofol concentration, on the basis of each of the five pharmacokinetic parameter sets. Then, for each of the five pharmacokinetic parameter sets, the performance in the population was determined by the median absolute performance error (MDAPE), the median performance error (MDPE), the wobble (the median absolute deviation of each PE from the MDPE), and the divergence (the percentage change of the absolute PE with time). The MDPE and MDAPE were compared between the parameter sets by the multisample median test. The initially used pharmacokinetic parameter set from Gepts et al. resulted in a MDPE of 24% and MDAPE of 26%.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7486117 TI - Awake endotracheal intubation in patients with cervical spine disease: a comparison of the Bullard laryngoscope and the fiberoptic bronchoscope. PMID- 7486119 TI - Posterior hypothalamic noradrenaline release during emergence from sevoflurane anesthesia in rats. PMID- 7486120 TI - Nonlinear measures of heart rate variability after fentanyl-based induction of anesthesia. PMID- 7486121 TI - Perioperative management of aneurysmal subarachnoid hemorrhage: Part 2. Postoperative management. PMID- 7486118 TI - Venous air embolism and cervical microdiscectomy. PMID- 7486122 TI - Latex anaphylaxis: another case, another cause. PMID- 7486123 TI - Sudden increase in end-tidal nitrogen due to changing vaporizers. PMID- 7486124 TI - Dantrolene treatment of opioid-induced myoclonus. PMID- 7486125 TI - Opisthotonus and thiopental. PMID- 7486126 TI - Oxygen therapy in the postanesthesia care unit. PMID- 7486127 TI - Arterial carbon dioxide tensions during fiberoptic tracheal intubation: a comparison of the split laryngeal mask with the Berman II airway. PMID- 7486128 TI - Another use for the nerve stimulator? PMID- 7486129 TI - Digital sensory loss without pulse oximeter malfunction. PMID- 7486130 TI - Accidental prolonged mydriasis in anesthesiologists. PMID- 7486131 TI - Endotracheal tube failure: undetected by routine testing. PMID- 7486132 TI - Gram . meter does not accurately represent the unit for ventricular work. PMID- 7486133 TI - Intravenous therapy and outpatient surgery outcomes. PMID- 7486134 TI - Techniques using the Bullard laryngoscope. PMID- 7486135 TI - The pharmacodynamics and pharmacokinetics of alcuronium. PMID- 7486136 TI - On the new nasogastric tube with balloon for prevention of pulmonary aspiration of gastric contents. PMID- 7486137 TI - The effects of epidural morphine and epidural butorphanol on maternal outcomes after cesarean delivery. PMID- 7486138 TI - Different onset times of succinylcholine-induced neuromuscular blockade at the adductor pollicis and stapedius muscles. PMID- 7486140 TI - A method to improve clinical assessment of neuromuscular recovery. PMID- 7486139 TI - Spinal versus epidural anesthesia for cesarean section. PMID- 7486141 TI - Rapid correction of severe hyponatremia to normal values is dangerous. PMID- 7486142 TI - Sevoflurane: introduction and overview. PMID- 7486143 TI - Cardiovascular responses to sevoflurane: a review. AB - In conclusion, sevoflurane appears to be similar to isoflurane and desflurane with a few exceptions. Sevoflurane was not associated with increases in heart rate in adult patients and volunteers, whereas higher MACs of isoflurane and desflurane and rapid increases in the inspired concentrations of these two anesthetics have been associated with tachycardia. Increasing concentrations of sevoflurane progressively decrease blood pressure in a manner similar to the other volatile anesthetics, and in unstimulated volunteers this decrease may be slightly less than with isoflurane at a higher MAC. Sevoflurane appears similar to isoflurane in its effect on regional blood flows, including the hepatic, renal, and cerebral circulation. In animals, sevoflurane appears to be a slightly less potent coronary vasodilator than isoflurane, and in a dog model, sevoflurane has not been associated with coronary flow redistribution ("steal"). Sevoflurane decreases myocardial contractility in a manner similar to equianesthetic concentrations of isoflurane and desflurane, and does not potentiate epinephrine induced cardiac arrhythmias. Sevoflurane reduces baroreflex function in a manner similar to other volatile anesthetics. In several multicenter studies where patients with CAD or patients at high risk for CAD were randomized to receive either sevoflurane or isoflurane for cardiac or noncardiac surgery, the incidence of myocardial ischemia, infarction, and cardiac outcomes did not differ between treatment groups. Thus, sevoflurane has not been associated with untoward cardiovascular changes in volunteers and patients undergoing elective surgery compared with other volatile anesthetics, and it appears to offer a more stable heart rate profile than either isoflurane or desflurane. PMID- 7486144 TI - The ventilatory effects of sevoflurane. PMID- 7486145 TI - Biotransformation of sevoflurane. AB - Several characteristics of sevoflurane biotransformation are apparent from the preceding investigations. Metabolism is rapid, with fluoride and HFIP appearing in plasma within minutes after the start of sevoflurane administration (38 40,51). Peak plasma fluoride concentrations generally occur within approximately 1 h after the termination of sevoflurane administration in most patients, regardless of the dose or duration of exposure (ranging from 0.35-9.5 MAC-h) (39,48). Peak plasma inorganic fluoride concentrations are proportional to sevoflurane dose, measured in MAC-h (42-44). Inorganic fluoride concentrations decline rapidly after termination of sevoflurane administration, with concentrations well below peak levels by the first postoperative day. HFIP is rapidly conjugated, with more than 85% circulating in plasma as the glucuronide. Plasma HFIP concentrations peak later than fluoride concentrations, but both metabolites are eliminated at similar rates (52). Metabolism of sevoflurane does not contribute to the termination of clinical drug effect (52), unlike more extensively metabolized drugs such as halothane (55). Sevoflurane is metabolized by P-450 2E1, so pathophysiologic factors and drug interactions altering P-450 2E1 activity will also influence sevoflurane metabolism (52). The extent of metabolism of sevoflurane, 2% to 5%, is less than that of all other volatile anesthetics except isoflurane and desflurane. It has been proposed that the ideal anesthetic should resist biotransformation because anesthetic toxicity is related to anesthetic metabolism (67,68). Experience to date suggests that biotransformation of sevoflurane has not been causally related to either hepatic or renal toxicity. Sevoflurane does not result in formation of fluoroacetylated liver neoantigens or other reactive metabolites. Although both sevoflurane and methoxyflurane may produce plasma fluoride concentrations in excess of 50 microM, they have not produced the same nephrotoxic effects. Clearly, anesthetic metabolism and anesthetic toxicity can no longer be considered synonymous. The introduction of sevoflurane into clinical practice will hopefully stimulate new investigations into biochemical mechanisms of anesthetic toxicity and continued clinical investigations regarding the relationship between anesthetic metabolism and organ toxicity. PMID- 7486146 TI - Sevoflurane and renal function. PMID- 7486147 TI - Sevoflurane in pediatric anesthesia. PMID- 7486148 TI - The hepatic effects of sevoflurane. PMID- 7486149 TI - The organ toxicity of inhaled anesthetics. PMID- 7486150 TI - The role of sevoflurane in outpatient anesthesia. AB - Sevoflurane appears to have several properties that make it an attractive alternative to the currently available anesthetics for outpatient anesthesia. The relative low solubility of sevoflurane, as well as an impressive lack of airway irritation, makes it a very useful anesthetic for inhalation induction of anesthesia. This feature is likely to make sevoflurane a population choice for pediatric outpatient anesthesia. For adult outpatients, the relatively low solubility of sevoflurane permits rapid alterations in alveolar concentration during the maintenance period (even when administered at relatively low flow rates of 2-3 L/min), thereby improving control of the depth of anesthesia. Low solubility should also allow for a more predictable emergence from anesthesia (versus isoflurane), thereby facilitating patient turnover in a busy ambulatory surgery center. The commercial development of sevoflurane has been slow because of concerns regarding the potential toxicity of its metabolites and breakdown products (24). However, the large body of clinical experience from Japan and the worldwide Phase II and III clinical development programs suggests that these concerns may be unfounded. The short duration of exposure in the outpatient setting should further decrease the risk of nephrotoxicity. The relatively low solubility of sevoflurane will facilitate its use with total gas flow rates of 2 3 L/min. In the final analysis, clinicians will have to balance the cost of sevoflurane (versus halothane, enflurane, isoflurane, and desflurane) against its potential advantages in the ambulatory surgery population. Although the search for anesthetics that are more ideally suited for use in the outpatient setting will continue, sevoflurane clearly represents a step in the right direction (3). PMID- 7486151 TI - Arteriovenous differences in plasma concentrations of catechols in rats with neuropathic pain. AB - BACKGROUND: Alterations in cutaneous temperature, sweating, and cutaneous blood flow in patients with pain states, such as reflex sympathetic dystrophy and causalgia, have been interpreted as evidence for exaggerated sympathetic outflow. It was determined whether pain behavior in a rat model of sympathetically maintained pain is associated with alterations in regional sympathoneural function. METHODS: Peripheral neuropathy was induced in 29 Sprague-Dawley rats by ligation of the left L5 and L6 spinal nerves. Sixteen other rats had sham surgery (nerve exposure without ligation). Animals were tested for behavioral signs of allodynia (decreased paw withdrawal thresholds to mechanical stimuli) at 2 and 4 weeks after the surgery. Arterial and iliac venous blood samples (left, affected; right, control) were obtained at 2 weeks (NP2, n = 14) and 4 weeks (NP4, n = 15) after neuropathic or sham (n = 8 at 2 and 4 weeks) surgery. Plasma concentrations of dihydroxyphenylalanine, dihydroxyphenylacetic acid, dopamine, norepinephrine, and the intraneuronal norepinephrine metabolite, 3,4-dihydroxyphenylglycol, were analyzed in arterial and left and right iliac venous samples. RESULTS: A decrease in paw withdrawal threshold was observed in neuropathic (NP2 and NP4) but not sham-operated rats. Affected and control limbs did not differ in arteriovenous differences in concentrations of dihydroxyphenylalanine, dihydroxyphenylacetic acid, dopamine, or 3,4-dihydroxyphenylglycol. No differences were observed between sham-operated and neuropathic animals in these arteriovenous increments. In contrast, affected limbs of NP2 rats had a reduced arteriovenous increment in norepinephrine concentrations, compared to that in the control side (P < 0.05). CONCLUSIONS: No neurochemical evidence of sympathetic hyperactivity is observed in the rat model of neuropathic pain; if anything, norepinephrine release is decreased in the affected limb. Autonomic disturbances in neuropathic pain are therefore more likely the result of receptor supersensitivity than increased local sympathoneural traffic. PMID- 7486152 TI - Effects of midazolam on intracellular Ca2+ and tension in airway smooth muscles. AB - BACKGROUND: It has been demonstrated that a group of minor tranquilizers, benzodiazepines, are able to relax airway smooth muscles. To determine the underlying mechanisms of this phenomenon, the effects of midazolam on the intracellular Ca2+ concentration ([Ca2+]i) and tension in airway smooth muscles were investigated. METHODS: Using front-surface fluorometry and fura-2-loaded porcine tracheal smooth muscle strips, both [Ca2+]i and isometric tension developments were simultaneously recorded. RESULTS: When the tracheal strips were exposed to a high external K(+)-solution (40 mM) or 10(-7) M carbachol containing 1.25 mM Ca2+, both [Ca2+]i and tension increased rapidly until they reached a plateau (the steady state). During steady-state contraction induced by K(+) depolarization or carbachol, the cumulative application of midazolam (10(-7) approximately 10(-4) M) caused decreases in both [Ca2+]i and tension, in a concentration-dependent manner. During 40 mM K(+)-induced depolarization, the stepwise increases in the extracellular Ca2+ concentration induced the stepwise increases in [Ca2+]i and tension. Midazolam (3 x 10(-5) M) inhibited these increases in [Ca2+]i and tension, but had no effect on the [Ca2+]i-tension relationship. In the presence of 3 x 10(-3) M NiCl2 (a nonselective cation channel blocker), midazolam (3 x 10(-5) M) did not cause any additional reduction of [Ca2+]i or tension during the contraction induced by carbachol (10(-7) M). In the absence of extracellular Ca2+, midazolam (3 x 10(-5) M) had no effect on the transient increases in either [Ca2+]i or the tension induced by carbachol (10(-7) M) or caffeine (20 mM). Pretreatment with both 10(-5) M flumazenil (a specific central antagonist of benzodiazepines) and 10(-5) M PK11195 (a specific peripheral antagonist of benzodiazepines) did not influence the effect of 10(-5) M midazolam on [Ca2+]i or tension during the contractions induced by carbachol. CONCLUSIONS: Midazolam directly relaxes airway smooth muscles by decreasing [Ca2+]i; this can be attributed to the inhibition of the influx of extracellular Ca2+. Midazolam has no effect on the release of stored Ca2+. In addition, midazolam has no effect on Ca2+ sensitivity of the contractile apparatus. Finally, benzodiazepine antagonists, flumazenil and PK11195, have no effect on this mechanism of direct action of midazolam on airway smooth muscles. PMID- 7486153 TI - Desflurane and isoflurane exert modest beneficial actions on left ventricular diastolic function during myocardial ischemia in dogs. AB - BACKGROUND: Volatile anesthetics exert cardioprotective effects during myocardial ischemia. This investigation examined the regional systolic and diastolic mechanical responses to brief left anterior descending coronary artery (LAD) occlusion in the central ischemic zone and in remote normal myocardium in the conscious state and during desflurane and isoflurane anesthesia. METHODS: Eighteen experiments were performed in nine dogs chronically instrumented for measurement of aortic and left ventricular pressure, cardiac output, LAD coronary blood flow velocity, and LAD and left circumflex coronary artery subendocardial segment length. Regional myocardial contractility was evaluated with the slope of the preload recruitable stroke work relationship determined from a series of left ventricular pressure-segment length diagrams in the LAD and left circumflex coronary artery zones. Diastolic function was assessed with a time constant of isovolumic relaxation (tau), maximum segment lengthening velocity in LAD and left circumflex coronary artery regions, and regional chamber stiffness constants derived using monoexponential and three-element exponential curve fitting in each zone. On separate experimental days, hemodynamics and indices of regional functional were obtained in the conscious state and during 1.1 and 1.6 minimum alveolar concentration end-tidal desflurane or isoflurane before and during LAD occlusion. RESULTS: In conscious dogs, LAD occlusion abolished regional stroke work, increased chamber stiffness (monoexponential: 0.39 +/- 0.04 during control to 1.34 +/- 0.39 mm-1 during LAD occlusion), and decreased the rate of early ventricular filling in the ischemic zone. These changes were accompanied by increased contractility (slope: 103 +/- 8 during control to 112 +/- 7 mmHg during LAD occlusion), rapid filling rate (maximum segment lengthening velocity: 46 +/- 5 during control to 55 +/- 7 mm.s-1 during LAD occlusion), and chamber stiffness (monoexponential: 0.43 +/- 0.05 during control to 1.14 +/- 0.25 mm-1 during LAD occlusion) in the normal region. Increases in tau were also observed in the conscious state during the period of myocardial ischemia. Desflurane and isoflurane increased tau and decreased the slope and maximum segment lengthening velocity in a dose-related manner. Monoexponential and three-element element exponential curve fitting were unchanged by the volatile anesthetics in the absence of ischemia. Myocardial contractility and rapid filling rate were enhanced in the nonischemic region during LAD occlusion in the presence of desflurane and isoflurane. In contrast to the findings in the conscious state, ischemia-induced increases in tau and chamber stiffness in the ischemic and normal zones were attenuated during anesthesia induced by desflurane and isoflurane. CONCLUSIONS: The results indicate that increases in contractility of remote myocardium during brief regional ischemia were preserved in the presence of desflurane and isoflurane anesthesia. In addition, desflurane and isoflurane blunted ischemia-induced increases in tau and regional chamber stiffness in both the ischemic and nonischemic zones. These results demonstrate that the volatile anesthetics may exert important beneficial actions on left ventricular mechanics in the presence of severe abnormalities in systolic and diastolic function during ischemia. PMID- 7486154 TI - Intrathecal amitriptyline. Antinociceptive interactions with intravenous morphine and intrathecal clonidine, neostigmine, and carbamylcholine in rats. AB - BACKGROUND: Systemically administered opioids induce analgesia in part by spinal noradrenergic, serotonergic, and cholinergic mechanisms. The current study tested whether antinociception from systemically administered opioids could therefore be enhanced by intrathecal injection of a monoamine reuptake inhibitor to potentiate the action of spinally released norepinephrine and serotonin (amitriptyline) and intrathecal injection of a cholinesterase inhibitor to potentiate the action of spinally released acetylcholine (neostigmine). METHODS: Rats were prepared with chronic lumbar intrathecal and femoral intravenous catheters and nociceptive threshold was assessed by hind paw withdrawal to a radiant heat stimulus. An isobolographic design was used to distinguish between additive and synergistic interactions. RESULTS: Intravenous morphine and intrathecal neostigmine, but not intrathecal amitriptyline, caused dose-dependent antinociception alone. Combining any two of these three treatments yielded a synergistic interaction compared to each alone, whereas combining all three yielded an additive interaction compared to each two-way interaction. Intrathecal amitriptyline did not affect antinociception from intrathecal clonidine or intrathecal carbamylcholine. CONCLUSIONS: These data suggest that intrathecal doses of amitriptyline resulting in potentiation of intravenous morphine antinociception may not be adequate to block muscarinic receptors, because they did not affect carbamylcholine-induced antinociception. These results further support the relevance of spinal monoamine reuptake and cholinesterase inhibition to synergistically enhance analgesia from systemic opioids. PMID- 7486156 TI - Analysis of halothane effects on myocardial force-interval relationships at anesthetic concentrations depressing twitches but not tetanic contractions. AB - BACKGROUND: Tetanic contractions in rat myocardium depend solely on cellular Ca2+ uptake, whereas twitches depend on Ca2+ release from the sarcoplasmic reticulum. Because halothane may cause loss of sequestered Ca2+, the anesthetic was tested for its differential effects on twitch and tetanic forces. The in vitro effects of halothane on the twitch force-interval relationship were then evaluated, using a mathematical model that relates twitch contractile force to the Ca2+ content of intracellular compartments. METHODS: Isometric contractile force was measured in paced (0.4 Hz) rat atrial preparations. The sarcoplasmic reticulum was functionally eliminated using ryanodine (10(-6) M), abolishing twitches. Rapid pacing (20 Hz, 10 s) caused tetanic contractions. The effects of identical halothane exposures on twitches and tetanic contractions were compared. Ca2+ compartment model parameters were extracted from twitch force-interval data, according to a previously employed quantitative procedure. RESULTS: Halothane (0.5-1%) depressed normal twitches, but not tetanic contractions. The anesthetic decreased the amplitude of the steady-state twitch force-frequency relationship, and accelerated the course of mechanical recovery. Halothane (0.5-1%) also accelerated the decay constant for the decline in amplitude of a series of rest potentiated contractions. The modeling showed that a 20-30% decrease in the recirculating fraction of activator Ca2+ accounts for 0.5% halothane-induced negative inotropy and acceleration of the decay constant. CONCLUSIONS: The differential effect of halothane on twitches and tetanic contractions implies that a functioning sarcoplasmic reticulum is required for halothane-induced negative inotropy. The effects of halothane on the force-interval relationship suggest that halothane reduces the sequestered pool of activator Ca2+. PMID- 7486157 TI - One-day hypothermic preservation of isolated hearts with halothane improves cardiac function better than low calcium. AB - BACKGROUND: Halothane exerts a potent negative inotropic effect on the heart and mimics many of the cardiac effects of lowered extracellular CaCl2. Reduced slow inward Ca2+ current and sarcoplasmic reticular effects on intracellular Ca2+ are likely involved. The authors reported previously that halothane protects against hypoxic and ischemia reperfusion injury in isolated hearts. The aim of this isolated heart study was to compare protective effects of halothane and low CaCl2 (0.5 mM) administered during 1 day of hypothermic perfusion on return of normothermic perfusion. METHODS: Guinea pig hearts (n = 66) were isolated and perfused at 37 degrees C with a Krebs' solution, gassed with 96% O2, 4% CO2, and containing 2.5 mM Ca2+, and 4.5 mM K+. Heart rate, isovolumetric left ventricular pressure, coronary flow, %O2 extraction, O2 consumption rate, and relative cardiac efficiency (EFF = heart rate.left ventricular pressure/O2 consumption rate) were measured in five groups of hearts: time controls (no hypothermia); 1.5, and 3% halothane delivered by vaporizer; cold controls (hypothermia only); and 0.5 mM CaCl2. Halothane was administered, or CaCl2 was decreased 0.5 h before hypothermia at 3.8 +/- 0.1 degrees C, during hypothermia for 22 h, and for 0.5 h after rewarming to 37.0 +/- 0.1 degrees C. Hearts were perfused at 25% of initial coronary flow during hypothermia. RESULTS: All groups had similar ventricular function and vasodilator responses before hypothermia. During normothermic reperfusion after hypothermia, both concentrations of halothane protected better than low CaCl2. Values, expressed as a percent of initial values in the five groups (time control, 3% halothane, 1.5% halothane, cold control, and 0.5 mM CaCl2, were respectively: 90 +/- 6, 54 +/- 6*, 48 +/- 5*, 27 +/- 8, 27 +/- 4% for left ventricular pressure; 84 +/- 5, 61 +/- 4*, 62 +/- 6*, 40 +/- 5, 34 +/- 5% for EFF; and 102 +/- 3, 63 +/- 3*, 66 +/- 3*, 55 +/- 2, 42 +/- 2% for coronary flow (*P < 0.05 halothane vs. 0.5 mM CaCl2). The coronary flow response to endothelium-dependent (acetylcholine) and endothelium-independent (nitroprusside) vasodilators was also greater after halothane than after 0.5 mM CaCl2. CONCLUSIONS: Halothane administered during hypothermia restores left ventricular pressure, cardiac efficiency, basal coronary flow, and flow responses better than low CaCl2. Although halothane and low CaCl2 both reduce intracellular Ca2+, contractile force, and metabolic demand, the better protective effect of halothane is not likely simply due to a reduction in contractile function and metabolic rate before or initially after hypothermia because these were reduced much more by low CaCl2 than by halothane. PMID- 7486155 TI - Intrathecal amitriptyline acts as an N-methyl-D-aspartate receptor antagonist in the presence of inflammatory hyperalgesia in rats. AB - BACKGROUND: Amitriptyline and other tricyclic antidepressants exhibit high affinity binding to N-methyl-D-aspartate (NMDA) receptors in vitro and inhibit NMDA receptor activation-induced neuroplasticity in hippocampal slices. Because spinal NMDA receptor activation is believed to be central to generation and maintenance of hyperalgesic pain, the purpose of this study was to test whether intrathecal amitriptyline reduced inflammation-induced hyperalgesia in the rat. METHODS: Rats were prepared with chronic lumbar intrathecal and femoral intravenous catheters and nociceptive threshold was assessed by hind paw withdrawal to a radiant heat stimulus. Rats received an injection of carrageenin in one hind paw followed by thermal paw withdrawal testing 3 hr later and intrathecal amitriptyline and/or intravenous morphine injection. In other rats, intrathecal NMDA injection was preceded by either intrathecal saline or 60 micrograms amitriptyline. RESULTS: Intrathecal amitriptyline reversed thermal hyperalgesia in a dose-dependent manner, but had no effect on withdrawal latency of the contralateral, noninjected paw. Intrathecal phentolamine plus methysergide did not alter amitriptyline's effect, except at the lowest dose. Intravenous morphine increased paw withdrawal latency in both inflamed and control paws in a dose-dependent fashion, and morphine interacted additively with intrathecal amitriptyline to reverse hyperalgesia. Thermal hyperalgesia induced by NMDA was completely antagonized by intrathecal amitriptyline. CONCLUSIONS: Amitriptyline and other tricyclic antidepressants have been demonstrated to exhibit modest activity against clinical neuropathic pain after systemic administration. These data suggest that more profound pain relief might be obtained by intrathecal administration. Amitriptyline reverses hyperalgesia in rats by a mechanism unrelated to monoamine reuptake inhibition, and likely due to NMDA receptor antagonism. PMID- 7486160 TI - Acute pain management. Programs in U.S. hospitals and experiences and attitudes among U.S. adults. AB - BACKGROUND: The objective of the survey was to assess the status of acute pain management in U.S. hospitals and attitudes of adults in the U.S. toward postoperative pain management, information that has not been previously available. METHODS: Two telephone questionnaire surveys were conducted U.S. hospital participants, including 100 teaching hospitals (acute care hospitals with a residency program and/or university affiliation), 100 nonteaching (community) hospitals with fewer than 200 beds, and 100 nonteaching (community) hospitals with 200 beds or more were interviewed regarding current and future pain management programs and related topics. Adult participants in 500 U.S. households were interviewed on attitudes and experiences with postoperative pain and its management. RESULTS: Forty-two percent of the hospitals have acute pain management programs, and an additional 13% have plans to establish an acute pain management program. Seventy-seven percent of adults believe that it is necessary to experience some pain after surgery. Fifty-seven percent of those who had surgery cited concern about pain after surgery as their primary fear experienced before surgery. Seventy-seven percent of adults reported pain after surgery, with 80% of these experiencing moderate to extreme pain. CONCLUSIONS: Despite a growing trend in pain management, increased professional and public awareness including the establishment of pain management programs and public and patient education is needed to reduce the incidence and severity of postoperative pain. PMID- 7486158 TI - Epinephrine is metabolized by the spinal meninges of monkeys and pigs. AB - BACKGROUND: Epinephrine commonly is added to epidural opioids and local anesthetics, however, little is known about the fate of epidurally administered epinephrine. Studies have identified the epinephrine metabolizing enzyme, catechol-O-methyl transferase (COMT), in the cranial meninges of several species. The purpose of this study was to determine whether the spinal meninges also contain COMT and are capable of metabolizing epinephrine. If so, then the spinal meninges may have an important impact in limiting the bioavailability of epinephrine in both the spinal cord and epidural space. METHODS: Spinal meningeal specimens measuring 4 cm2 were obtained from monkeys (M. nemestrina) and farm bred pigs and were incubated in bicarbonate-buffered mock cerebrospinal fluid. Epinephrine (200 micrograms base) was added at t = 0, and 200 min later, the mock cerebrospinal fluid was collected for metanephrine analysis. In separate experiments, pig meningeal specimens were separated into dura mater, pia arachnoid mater, and pia mater, and the experiments were repeated to determine which meninx had the greatest COMT activity. RESULTS: Metanephrine was produced by monkey meninges at the rate of 0.47 ng.min-1.cm-2 and by pig meninges at the rate of 0.23 ng.min-1.cm-2 (P > 0.05). The pia-arachnoid meninx produced metanephrine at a greater rate (4.48 +/- 0.46 ng.min-1.mg-1 tissue) than did the pia mater (1.3 +/- 0.15 ng.min-1.mg-1 tissue) or dura mater alone (1.82 +/- 0.23 ng.min-1.mg-1 tissue). CONCLUSIONS: These data demonstrate the functional presence of COMT in the spinal meninges of pigs and monkeys and suggest that the spinal meninges may limit the spinal bioavailability of epidurally or intrathecally administered epinephrine. PMID- 7486161 TI - Technique for quantifying the duration of intravenous anesthetic effect. AB - BACKGROUND: Several recent studies have analyzed the relationship between pharmacokinetic parameters and the rate of decrease in concentration after discontinuation of a continuous drug infusion. Although these studies have clarified our understanding of those aspects of pharmacokinetics most relevant to anesthesia practice, they do not directly address the issue of the duration of drug effect, which will be a function of both pharmacokinetic and pharmacodynamic variables. This paper extends these concepts by presenting a method to unify pharmacokinetics and pharmacodynamics in a measure of duration of drug effect that is applicable when the drug effect is assessed in a binary, response/no response fashion. METHODS: The parameter proposed to quantify duration of drug effect is the area under the curve expressing probability of drug effect as a function of time after the agent is discontinued. This parameter is denoted the mean effect time. It is calculated using the logistic (or Hill) equation to relate the probability of drug effect to drug concentration, which in turn can be calculated as a function of time by pharmacokinetic simulation. Mean effect times were calculated for sufentanil, alfentanil, propofol, and midazolam using the logistic equation describing recovery and by assuming that drug blood concentrations during maintenance of anesthesia were sufficient to reduce the probability of responsiveness to surgical stimulation to 10% (C90). Published pharmacokinetic and pharmacodynamic parameters were used for these calculations. These results were compared to the relevant decrement times (as defined in this paper, the time required for the concentration to decrease from C90 to the concentration at which 50% of patients are responsive and/or able to maintain adequate ventilation, denoted C50). It was assumed that C90 and C50 were independent variables. RESULTS: Mean effect times for midazolam and propofol, for which the steepness parameter delta for recovery (responsiveness and adequate ventilation) is less than 4, are significantly greater than the decrement time. Mean effect times for sufentanil and alfentanil (delta = 6 and 10, respectively) are close to decrement times. The discrepancy between mean effect time and decrement time becomes greater as the duration of drug administration increases. The incorporation of pharmacokinetic variability into the calculations had little effect on the results. CONCLUSIONS: Context-sensitive half-times or other decrement times have been shown to be the most useful measures of the kinetics of drug concentrations. Mean effect time may be a useful concept for understanding the recovery from drug effects. PMID- 7486162 TI - Evaluation of a double-lumen multiorifice catheter for resuscitation of swine from lethal venous air embolism. AB - BACKGROUND: A double-lumen multiorifice catheter has been developed to potentially enhance accurate electrocardiographic central venous localization and resuscitation from a massive venous air embolism (VAE). This double-lumen multiorifice catheter was compared to a Bunegin-Albin multiorifice catheter for flow characteristics, air aspiration efficiency, and efficacy in resuscitating swine from a lethal VAE. METHODS: Flow characteristics of both catheters were determined by aspirating both agitated and unagitated citrated swine blood with a 50-ml syringe. Swine were anesthetized with halothane and positioned to approximate a modified sitting craniotomy position (45-degree elevation). By a random block method, 24 swine were assigned to either catheter (n = 12 each catheter) for the initial air aspiration. Catheters were positioned, using intravenous electrocardiography, with the distal aspiration orifice in the high right atrium. A 5-ml/kg air embolism was administered over 30 s into the sagittal sinus, and the swine were resuscitated by aspirating air through the multiorifice catheters and then positioning the swine horizontally. Surviving animals were allowed to recover for 60 min. The initial catheter was exchanged and repositioned in the high right atrium using intravenous electrocardiography. A 5 ml/kg air embolus was administered, and the swine were resuscitated as in the first challenge. Surviving swine recovered for 60 min, repositioned, and administered a third 5-ml/kg air embolism. On this final challenge, no attempt was made to resuscitate the animal by aspirating the multiorifice catheter. RESULTS: Flow characteristics of both catheters were similar in the unagitated blood (195.3 +/- 1.9 vs. 196.7 +/- 2.5 ml/min). The flow rate of agitated blood through the double-lumen multiorifice catheter was 14% greater than through the Bunegin-Albin catheter (136.3 +/- 6.8 vs. 117 +/- 5.9 ml/min, P = 0.001). Forty three air embolism trials were conducted at 5 ml/kg. All nine trials at 5 ml/kg without air aspiration resulted in death. Five animals died during the embolism dose determination trials, and four died during the third embolism challenge. The use of a multiorifice catheter for aspiration after a VAE enhanced survival after a 5-ml/kg sagittal sinus air embolus (14/34 vs. 0/9, P = 0.02). Although the double-lumen multiorifice catheter was more efficient than the Bunegin-Albin catheter in percentage of air retrieved (37.7 +/- 12.0 vs. 29.7 +/- 10.1, P = 0.042). Aspiration of the VAE with the double-lumen multiorifice catheter successfully rescued 9 of the 15 trials, and aspiration using the Bunegin-Albin catheter resuscitated 5 of the 19 (P = 0.08). CONCLUSIONS: Multiorifice catheters are effective in resuscitating swine from a lethal VAE. The double-lumen multiorifice catheter evaluated aspirated a larger percentage of the VAE but was not statistically more effective than the Bunegin-Albin catheter in resuscitating the animals. Based on these findings of improved flow rate and efficiency in air aspiration, further investigation of this double-lumen multiorifice catheter is warranted. PMID- 7486159 TI - Further characterization of the receptor mechanism involved in the antidysrhythmic effect of dexmedetomidine on halothane/epinephrine dysrhythmias in dogs. AB - BACKGROUND: alpha 2 Adrenoceptors in the central nervous system mediate various physiologic processes, including cardiovascular control. Recently, some of these actions have been reported to be mediated by a nonadrenergic receptor, namely an imidazoline receptor. The authors previously reported that dexmedetomidine, a selective alpha 2 agonist, prevents the genesis of halothane-epinephrine dysrhythmias through a central mechanism. Because dexmedetomidine also binds to imidazoline receptors, we performed the current study to examine the precise receptor mechanism involved in the antidysrhythmic property of dexmedetomidine. METHODS: Adult mongrel dogs were anesthetized with halothane (1.3%) and monitored continuously for systemic arterial pressure and premature ventricular contractions. The dysrhythmogenic dose of epinephrine was defined as the smallest dose producing four or more premature ventricular contractions within 15-s period. We examined the antidysrhythmic action of dexmedetomidine in the presence of two kinds of alpha 2 antagonists, that is, agents that label imidazoline receptors and exert a pharmacologic action through imidazoline receptors (idazoxan and atipamezole) and agents that are nonimidazoline compounds and are lacking in pharmacologic action through imidazoline receptors (rauwolscine and L 659,066). They were given cerebroventricularly. RESULTS: Idazoxan and atipamezole significantly inhibited the antidysrhythmic action of dexmedetomidine, whereas rauwolscine and L-659,066 did not. CONCLUSIONS: Because alpha 2 antagonists having imidazoline or imidazole structures inhibited the antidysrhythmic action of dexmedetomidine, and the inhibition produced by the non-imidazoline alpha 2 antagonists was not significant, imidazoline receptors in the central nervous system are more responsible for the antidysrhythmic action of dexmedetomidine than are alpha 2 adrenoceptors. PMID- 7486163 TI - Pediatric caudal epidural catheter sequestration: an unusual complication. PMID- 7486164 TI - Lumbar stenosis complicating retained epidural catheter tip. PMID- 7486165 TI - High sensory block after intrathecal sufentanil for labor analgesia. PMID- 7486166 TI - An unusual cutaneous presentation of reflex sympathetic dystrophy. PMID- 7486168 TI - More on nasotracheal intubation in children. PMID- 7486167 TI - Hepatotoxicity after desflurane anesthesia. PMID- 7486169 TI - Pulmonary artery catheter passing between the chordae tendineae of the tricuspid valve. PMID- 7486171 TI - Role of epidural anesthesia and analgesia on postoperative patient outcome. PMID- 7486172 TI - Should renaming the specialty begin with the Journal? PMID- 7486170 TI - Use of a left-sided double-lumen tube to occlude the right upper lobe orifice. PMID- 7486173 TI - Another simple method for ring removal. PMID- 7486174 TI - Preoperative risk assessment: many studies, few solutions. Is a cardiac risk assessment paradigm possible? PMID- 7486175 TI - Evolving clinically useful predictors of recovery from intravenous anesthetics. PMID- 7486176 TI - Preoperative dipyridamole thallium imaging and ambulatory electrocardiographic monitoring as a predictor of perioperative cardiac events and long-term outcome. AB - BACKGROUND: Dipyridamole thallium imaging (DTI) and ambulatory electrocardiography (AEGC) have been advocated as means to stratify risk before vascular surgery. The purpose of this study was to compare the predictive value of both tests in noncardiac surgery patients for perioperative cardiac morbidity and long-term mortality. METHODS: One hundred eighty patients were referred to the nuclear cardiology laboratory for DTI before noncardiac surgery. In patients with normal electrocardiograms and who consented, an ambulatory electrocardiogram was recorded for 24 h. DTI results were classified as negative, positive, or strongly positive (included in positive). Patients were assessed for a minimum of 12 months, and Kaplan-Meier cardiovascular survival curves were constructed with a log-rank statistic of equality with P < 0.05 significant. RESULTS: One hundred nine patients had both tests and then underwent surgery, sustaining 10 perioperative cardiac events (cardiac death, myocardial infarction, or symptomatic ischemia). The positive predictive values for DTI (18%) and AECG (25%) were similar, as were the likelihood ratios for positive tests (DTI = 2.1, AECG = 3.3). The likelihood ratios of a negative test were also similar (DTI = 0.45, AECG = 0.48). A strongly positive thallium defect had a somewhat greater likelihood ratio (3.5) for in-hospital events and was the only test result associated with a significantly worse long-term cardiac survival. CONCLUSIONS: AECG and DTI demonstrated a similar, although lower than initially reported, ability to stratify risk and predict short-term outcome. Only quantitative dipyridamole thallium also had predictive value for long-term prognosis. PMID- 7486177 TI - Cardiovascular effects of sevoflurane compared with those of isoflurane in volunteers. AB - BACKGROUND: Sevoflurane is a new inhalational anesthetic with desirable clinical properties. In some clinical situations, an understanding of the detailed cardiovascular properties of an anesthetic is important, so the authors evaluated the hemodynamic effects of sevoflurane in healthy volunteers not undergoing surgery. METHODS: Twenty-one subjects were randomized to receive sevoflurane, isoflurane, or sevoflurane: 60% N2O. Anesthesia was induced and maintained by inhalation of the designated anesthetic. Hemodynamic measurements were performed before anesthesia, during controlled ventilation, during spontaneous ventilation, and again during controlled ventilation after 5.5 h of anesthesia. RESULTS: A few subjects became excessively hypotensive at high anesthetic concentrations (2.0 minimum alveolar concentration [MAC] sevoflurane, 1.5 and 2.0 MAC isoflurane), preventing data collection. Sevoflurane did not alter heart rate, but decreased mean arterial pressure and mean pulmonary artery pressure. Cardiac index decreased at 1.0 and 1.5 MAC, but in subjects with mean arterial pressure > or = 50 mmHg returned to baseline values at 2.0 MAC when systemic vascular resistance decreased. Sevoflurane did not alter echocardiographic indices of ventricular function, but did decrease an index of afterload. Sevoflurane caused a greater decrease in mean pulmonary artery pressure than did isoflurane, but the cardiovascular effects were otherwise similar. Administration of sevoflurane with 60% N2O, prolonged administration or spontaneous ventilation resulted in diminished cardiovascular depression. CONCLUSIONS: At 1.0 and 1.5 MAC, sevoflurane was well tolerated by healthy volunteers. At 2.0 MAC, in subjects with mean arterial pressure > or = 50 mmHg, no adverse cardiovascular properties were noted. Similar to other contemporary anesthetics, sevoflurane caused evidence of myocardial depression. Hemodynamic instability was noted in some subjects at high anesthetic concentrations in the absence of surgical stimulation. The incidence was similar to that with isoflurane. The cardiovascular effects of sevoflurane were similar to those of isoflurane, an anesthetic commonly used in clinical practice since 1981. PMID- 7486178 TI - Dose-response curves of inhaled nitric oxide with and without intravenous almitrine in nitric oxide-responding patients with acute respiratory distress syndrome. AB - BACKGROUND: Inhaled nitric oxide, a selective pulmonary vasodilator, in combination with intravenous almitrine, a selective pulmonary vasoconstrictor, markedly improves arterial oxygenation in 50-60% of patients with acute lung injury. The goal of this study was to assess dose response of inhaled nitric oxide with and without almitrine in patients with acute respiratory distress syndrome responding to nitric oxide. METHODS: Six critically ill patients (aged 44 +/- 7 yr) were studied during early stage of their acute respiratory failure (Murray score: 2.6 +/- 0.1). All responded to 15 parts per million (ppm) of inhaled nitric oxide by an increase in Pao2 of at least 40 mmHg at FIo2 1. Hemodynamic and respiratory parameters were recorded continuously from pulmonary artery and systemic catheters. Inspiratory, expiratory, and mean intratracheal nitric oxide concentrations were monitored continuously using a fast response time chemiluminescence apparatus (NOX 4000, Seres, Aix-en-provence, France). On day 1, 6 inspiratory concentrations of nitric oxide were randomly administered: 0.15, 0.45, 1.5, 4.5, 15, and 45 ppm to determine the dose response of inhaled nitric oxide on Pao2, pulmonary shunt, mean pulmonary artery pressure, and pulmonary vascular resistance index. On day 2, a continuous intravenous infusion of almitrine at a dose of 16 micrograms.kg-1.min-1 was administered and dose response to inhaled nitrix oxide was repeated according to the same protocol as during day 1. A constant FIo2 of 0.85 was used throughout the study. RESULTS: Nitric oxide induced a dose-dependent increase in Pao2 for inspiratory nitric oxide concentrations ranging between 0.15 and 1.5 ppm. Almitrine increased Pao2/FIo2 from 161 +/- 30 to 251 +/- 45 mmHg (P < 0.001) and pulmonary vascular resistance index from 455 +/- 185 to 527 +/- 176 dyn.s.cm-5.m2 (P < 0.05), and decreased pulmonary shunt (Qs/QT) from 35 +/- 2 to 33 +/- 3% (P < 0.001). During almitrine combined with nitric oxide, a dose-dependent increase in Pao2 was observed for inspiratory nitric oxide concentrations ranging between 0.15 and 1.5 ppm. Almitrine plus nitric oxide 1.5 ppm increased Pao2/FIo2 from 161 +/- 30 to 355 +/- 36 mmHg (P < 0.001), decreased Qs/QT from 35 +/- 2 to 24 +/- 2% (P < 0.001), pulmonary vascular resistance index from 455 +/- 185 to 385 +/- 138 dyn.s.cm-5.m2 (P < 0.05), and mean pulmonary artery pressure from 31 +/- 4 to 28 +/- 4 mmHg (P < 0.001). CONCLUSIONS: In 6 patients with early acute respiratory distress syndrome and highly responsive to inhaled nitrix oxide, the administration of intravenous almitrine at a concentration of 16 micrograms.kg 1.min-1 induced an additional increase in Pao2. Dose response of nitric oxide was not changed by the administration of almitrine and a plateau effect was observed at inspiratory nitric oxide concentrations of 1.5 ppm. PMID- 7486179 TI - Pharmacokinetics of computer-controlled alfentanil administration in children undergoing cardiac surgery. AB - BACKGROUND: Cardiopulmonary bypass (CPB) induces changes in the pharmacokinetics of drugs. The purpose of this study was to model the pharmacokinetics of alfentanil in children undergoing cardiac surgery to provide accurate dosage titration intraoperatively as well as in the postoperative period. METHODS: Fourteen children (aged 3 months to 8 yr) undergoing cardiac surgery with CPB were administered alfentanil via a computer-controlled infusion pump. During surgery, the computer-controlled infusion pump was set to target plasma alfentanil concentrations of 500-2500 micrograms/ml. After surgery, the computer controlled infusion pump was set to target plasma concentrations of 200-500 micrograms/ml. Parameters for children previously published by Goresky et al. were programmed into the device. Arterial blood samples were taken throughout the infusion. Plasma samples were assayed by radioimmunoassay. Alfentanil pharmacokinetics were estimated using a pooled-data approach with a simple weight proportional, three-compartment mamillary model with parameters expressed in volumes and clearances as well as a CPB-adjusted, three-compartment model in which the parameters were allowed to change before, during, and after CPB. The accuracy of the three models was compared using cross-validation. RESULTS: Plasma alfentanil concentrations during computer-controlled infusion pump administration exceeded target concentrations for the first 10 min of drug administration, and from 300 min to the end of the study. The median absolute performance error was 33%. Pharmacokinetic modeling estimated a set of parameters for a simple three compartment model with a median absolute weighted residual of 18.4%. A CPB adjusted model nominally decreased the median absolute weighted residual to 17.0%. The performance of these models as measured by cross-validation performance was 18.9% median absolute performance error for the simple model and 18.4% median absolute performance error for the CPB-adjusted model. Parameters for the simple three-compartment model are: V1 = 19.2 ml.kg-1; V2 = 99 ml.kg-1; V3 = 2344 ml.kg-1; Cl1 = 2.5 ml.kg-1.min-1; Cl2 = 38 ml.kg-1.min-1; and Cl3 = 15 ml.kg-1.min-1. In the CPB-adjusted model V1, V2, and Cl2 changed with the onset of CPB. After CPB, V1 and Cl2 returned to the initial values, while V2 was described by a third value. CONCLUSIONS: The population pharmacokinetics of alfentanil in children undergoing cardiac surgery were well described by both a simple weight-proportional, three-compartment model and a weight-proportional, CPB-adjusted three-compartment model. Cross-validation estimated an expected median inaccuracy of approximately 18-20% with the estimated models in identical experimental circumstances. The flexible CPB-adjusted pharmacokinetic model could be used for modeling any drug with linear pharmacokinetics given in the context of CPB. PMID- 7486180 TI - Effect of antiemetic therapy on recovery and hospital discharge time. A double blind assessment of ondansetron, droperidol, and placebo in pediatric patients undergoing ambulatory surgery. AB - BACKGROUND: Postoperative nausea and vomiting continue to be a significant problem for pediatric ambulatory surgery patients. Although ondansetron has been demonstrated to be effective in the prophylactic treatment of postoperative nausea and vomiting (surrogate end point) no one has demonstrated a benefit of antiemetic therapy on patient recovery, postanesthesia care unit length of stay, and hospital length of stay (nonsurrogate end points). In a double-blind manner, the effects of ondansetron, droperidol, and placebo on the incidence of emesis, postanesthesia care unit stay, and hospital discharge time were evaluated in children undergoing dental surgery. METHODS: The subjects were 102 children aged 2-8 years undergoing complete dental restoration. All patients received midazolam before undergoing inhalation induction of anesthesia with N2O/O2 and halothane. Anesthesia was maintained with N2O/O2 and alfentanil. Patients were then randomized to receive ondansetron (0.1 mg/kg), droperidol (75 micrograms/kg), or placebo (normal saline) in a double-blind fashion. At the conclusion of the anesthesia, a trained nurse observer assessed patient recovery and recorded the time patients met specified criteria for postanesthesia care unit and hospital discharge as well as episodes of emesis in the hospital and at home during the first 24 hr after surgery. RESULTS: Ninety-five patients completed the study. The three antiemetic groups were similar with respect to age, weight, length of surgery, dose of alfentanil, and route of preanesthetic medication. The 24-hr incidence of emesis was significantly less with ondansetron (9%) than with placebo (35%) or droperidol (32%). Ondansetron-treated patients had significantly shorter hospital stays than droperidol-treated patients, but recovery parameters were similar between the ondansetron- and placebo-treated patients. CONCLUSIONS: Ondansetron is an effective prophylactic antiemetic agent for children undergoing dental surgery. Compared with droperidol, ondansetron decreases the length of hospital stay, but compared to placebo, there were no differences in the patient recovery parameters. PMID- 7486181 TI - Heat flow and distribution during epidural anesthesia. AB - BACKGROUND: Core hypothermia after induction of epidural anesthesia results from both an internal core-to-peripheral redistribution of body heat and a net loss of heat to the environment. However, the relative contributions of each mechanism remain unknown. The authors thus evaluated regional body heat content and the extent to which core hypothermia after induction of anesthesia resulted from altered heat balance and internal heat redistribution. METHODS: Twelve minimally clothed male volunteers were evaluated in a approximately 22 degrees C environment for 2.5 control hours before induction of epidural anesthesia and for 3 subsequent hours. Epidural anesthesia produced a bilateral sympathetic block in only six volunteers, and only their results are reported. Shivering, when observed, was treated with intravenous meperidine. Overall heat balance was determined from the difference between cutaneous heat loss (thermal flux transducers) and metabolic heat production (oxygen consumption). Arm and leg tissue heat contents were determined from 19 intramuscular needle thermocouples, 10 skin temperatures, and "deep" foot temperature. To separate the effects of redistribution and net heat loss, we multiplied the change in overall heat balance by body weight and the specific heat of humans. The resulting change in mean body temperature was subtracted from the change in esophageal or tympanic membrane (core) temperatures, leaving the core hypothermia specifically resulting from redistribution. RESULTS: Arm heat content decreased approximately 5 kcal/h after induction of anesthesia, but leg heat content increased markedly. Most of the increase in leg heat content was in the lower legs and feet. Core temperature increased slightly during the control period but decreased 0.8 +/- 0.3 degrees C in the 1st hour of anesthesia. Redistribution, contributing 89% to this initial decrease, required a net transfer of 20 kcal from the trunk to the extremities. During the subsequent 2 h of anesthesia, core temperature decreased an additional 0.4 +/- 0.3 degrees C, with redistribution contributing 62%. Thus, only 7 kcal were redistributed during the 2nd and 3rd hours of anesthesia. Redistribution therefore contributed 80% to the entire 1.2 +/- 0.3 degrees C decrease in core temperature during the 3 h of anesthesia. CONCLUSIONS: Core hypothermia during the 1st hour after induction of epidural anesthesia resulted largely from redistribution of body heat from the core thermal compartment to the distal legs. Even after 3 h of anesthesia, redistribution remained the major cause of core hypothermia. Despite the greater fractional contribution of redistribution during epidural anesthesia, core temperature decreased only half as much as during general anesthesia because metabolic rate was maintained and the arms remained vasoconstricted. PMID- 7486182 TI - Measured context-sensitive half-times of remifentanil and alfentanil. AB - BACKGROUND: The context-sensitive half-time, rather than the terminal elimination half-life, has been proposed as a more clinically relevant measure of decreasing drug concentration after a constant infusion of a given duration. The context sensitive half-time is derived from computer modelling using known pharmacokinetic parameters. The modelled context-sensitive half-time for a 3-h infusion of alfentanil is 50-55 min and is 3 min for remifentanil. The terminal elimination half-life is 111 min for alfentanil and 12-30 min for remifentanil. It has not been tested whether the modelled context-sensitive half-time reflects the true time for a 50% decrease in drug concentration or drug effect. METHODS: Thirty volunteers received a 3-h infusion of remifentanil or alfentanil at equieffective concentrations. Depression of minute ventilation to 7.5% ETCO2 was used as a measure of drug effect. Minute ventilation response was measured, and blood samples for drug concentration were taken during and after drug infusion. The recovery of minute ventilation (drug effect) and decrease in blood drug concentration was plotted, and the time for a 50% change was determined. RESULTS: The measured pharmacokinetic context-sensitive half-time for remifentanil after a 3-h infusion was 3.2 +/- 0.9 min, and its pharmacodynamic offset was 5.4 +/- 1.8 min. Alfentanil's measured pharmacokinetic context-sensitive half-time was 47.3 +/- 12 min, and its pharmacodynamic offset was 54.0 +/- 48 min. The terminal elimination half-life modelled from the volunteers was 11.8 +/- 5.1 min for remifentanil and 76.5 +/- 12.6 min for alfentanil. CONCLUSIONS: The measured context-sensitive half-times were in close agreement with the context-sensitive half-times previously modelled for these drugs. The results of this study confirm the value of the context-sensitive half-time in describing drug offset compared to the terminal elimination half-life. PMID- 7486183 TI - Faster recovery after anesthesia in infants after intravenous induction with methohexital instead of thiopental. AB - BACKGROUND: To determine possible delays in recovery after intravenous anesthesia induction with thiopental, the drug was compared with methohexital in infants 1 12 months of age who were scheduled for hernia repair or circumcision. METHODS: The infants were given equipotent doses of methohexital (3.0 mg/kg, n = 21) or thiopental (7.3 mg/kg, n = 20), in random and blind fashion. After tracheal intubation, anesthesia was maintained with isoflurane in nitrous oxide/oxygen. All children received 0.75 ml/kg caudal bupivacaine (2.5 mg/ml). Isoflurane was discontinued at the beginning of skin closure, and nitrous oxide was terminated immediately after the last suture (end of surgery). RESULTS: There were no differences between the two groups with respect to age, weight, or duration of surgery, which lasted 19 min (14-23 min) in the methohexital group and 16 min (15 19 min) in the thiopental group (median and inner quartile range). Time from termination of nitrous oxide to extubation did not differ significantly between the groups. Time to spontaneous eye opening after end of surgery was 23 min (5-44 min) after methohexital induction and 55 min (25-74 min) after thiopental induction (P < 0.05). Recovery, assessed as postanesthetic recovery scores by a blinded observer, was significantly more rapid in the methohexital group at arrival in the recovery room and 5, 15, and 45 min after arrival. After 120 min, almost all infants of both groups were awake. CONCLUSIONS: Recovery after short surgical procedures in infants is faster after intravenous induction with methohexital than with thiopental. PMID- 7486184 TI - Direct cerebrovasodilatory effects of halothane, isoflurane, and desflurane during propofol-induced isoelectric electroencephalogram in humans. AB - BACKGROUND: The effect of volatile anesthetics on cerebral blood flow depends on the balance between the agent's direct vasodilatory action and the indirect vasoconstrictive action mediated by flow-metabolism coupling. To compare the intrinsic action of volatile anesthetics, the effect of halothane, isoflurane, and desflurane on flow velocity in the middle cerebral artery during propofol induced isoelectricity of the electroencephalogram was examined. METHODS: In 21 ASA physical status 1-2 patients, anesthesia was induced with 2.5 mg/kg propofol, 3 micrograms/kg fentanyl, and 0.1 mg/kg vecuronium and maintained with a propofol infusion to preserve an isoelectric electroencephalogram. End-tidal carbon dioxide and blood pressure were maintained constant throughout the study period. A transcranial Doppler was used to measure blood flow velocity in the middle cerebral artery, and a catheter was inserted in a retrograde direction into the jugular bulb for oxygen saturation measurements. After 15 min of isoelectric electroencephalogram, arterial and jugular venous blood samples were drawn, and flow velocity in the middle cerebral artery was recorded. Patients were randomly allocated to receive 0.5 MAC halothane, isoflurane, or desflurane, and after 15 min of equilibration, all variables were measured again. The concentration of the volatile agent was increased to 1.5 MAC, and after 15 min of equilibration, the measurements were repeated. RESULTS: Halothane, isoflurane, and desflurane significantly increased flow velocity in the middle cerebral artery (baseline 28 +/- 4, 30 +/- 4, and 29 +/- 3 cm/s, respectively) at 0.5 MAC (19 +/- 1.5%, 21 +/- 2%, and 23 +/- 3%, respectively; P < 0.05) and at 1.5 MAC (48 +/- 3%, 75 +/- 7%, and 74 +/- 4%, respectively; P < 0.05). Changes in the cerebral arteriovenous oxygen content difference are consistent with these findings. CONCLUSIONS: Halothane, isoflurane, and desflurane have intrinsic, dose-related cerebral vasodilatory effects. Whereas all three agents are similar at 0.5 MAC, isoflurane and desflurane have greater vasodilatory effects than halothane at 1.5 MAC. PMID- 7486185 TI - Atracurium versus vecuronium in asthmatic patients. A blinded, randomized comparison of adverse events. AB - BACKGROUND: To determine which of atracurium or vecuronium is associated with fewer adverse cardiovascular and pulmonary events in high-risk patients, the authors administered these drugs to patients with known asthma. METHODS: Sixty patients aged 18-75 yr taking bronchodilators chronically for asthma were anesthetized with midazolam, fentanyl, nitrous oxide, and isoflurane; the trachea was intubated without paralysis. When anesthetic conditions and mechanical ventilation were stable, patients were randomly given 0.5 mg/kg atracurium or 0.1 mg/kg vecuronium over 5-10 s, and a blinded observer recorded cardiovascular, pulmonary, and cutaneous signs of adverse reactions for 6 min. RESULTS: Arterial pressures and heart rate decreased after atracurium, and systolic pressure and heart rate decreased with vecuronium; these changes were small in magnitude. Cardiovascular effects (decrease in blood pressure or change in heart rate) > 10% were common with both atracurium (60% of patients) and vecuronium (57%). Cardiovascular effects > 20% were more frequent with atracurium (37%) than with vecuronium (13%, P < 0.05). The incidence of noncardiovascular adverse events (increase in peak airway pressure > 5 cmH2O, tidal volume decrease > 10%, rashes, and wheezing) did not differ between atracurium (17%) and vecuronium (7%). The largest increase in peak airway pressure was 5.1 cmH2O in a patient whose tidal volume decreased 16% with vecuronium; in the remaining patients, tidal volume decreased < 10%. No patients experienced inspiratory wheezing, marked decreases in arterial oxygen saturation, or marked increases in end-tidal carbon dioxide tension. CONCLUSIONS: The authors conclude that, in patients with asthma, adverse cardiovascular events are more common with atracurium than with vecuronium. PMID- 7486186 TI - Role of the vagus nerve in the antidysrhythmic effect of dexmedetomidine on halothane/epinephrine dysrhythmias in dogs. AB - BACKGROUND: Dexmedetomidine, an alpha 2-adrenergic agonist, can prevent the genesis of halothane/epinephrine dysrhythmias through the central nervous system. Because stimulation of alpha 2 adrenoceptors in the central nervous system enhances vagal neural activity and vagal stimulation is known to inhibit digitalis-induced dysrhythmias, dexmedetomidine may exert the antidysrhythmic property through vagal stimulation. To address this hypothesis, the effect of dexmedetomidine in vagotomized dogs was examined and compared with that in intact dogs. In addition, the effect of vagotomy on the antidysrhythmic action of doxazosin, an alpha 1 antagonist, was studied. METHODS: Adult mongrel dogs were anesthetized with halothane (1.3%) and monitored continuously for systemic arterial pressure and premature ventricular contractions. Animals were divided into two groups receiving bilateral vagotomy or sham operation. The dysrhythmia threshold was expressed by the dysrhythmogenic dose of epinephrine, defined as the smallest dose producing four or more premature ventricular contractions within a 15-s period, and plasma concentration of epinephrine at the time when the dysrhythmogenic dose was reached. The threshold was determined in the presence of dexmedetomidine (a selective alpha 2 agonist that crosses the blood brain barrier) and doxazosin (a selective alpha 1 antagonist that does not penetrate the blood-brain barrier) in the two groups. In addition, the effect of dexmedetomidine in the presence of atropine methylnitrate instead of vagotomy was examined. RESULTS: Vagotomy did not affect the basal vulnerability to halothane/epinephrine dysrhythmias significantly. Although dexmedetomidine dose dependently prevented the genesis of the dysrhythmias in intact dogs, the beneficial effect of dexmedetomidine was abolished in both the vagotomized and the atropine-treated dogs. On the other hand, vagotomy did not change the antidysrhythmic property of doxazosin. CONCLUSIONS: The vagus nerve plays an important role in the prevention of halothane/epinephrine dysrhythmias by dexmedetomidine in dogs. However, resting vagal tone neither modulates the onset of halothane/epinephrine dysrhythmias nor affects the antidysrhythmic action of doxazosin. PMID- 7486187 TI - [Complex detoxication in crush syndrome]. PMID- 7486188 TI - [Features of the treatment of suppurative complications of limb injuries in prolonged crush syndrome]. AB - The results of treatment of 42 patients with prolonged crush syndrome complicated by suppuration are analyzed. The results of bacteriological studies are presented, which helped monitor the treatment efficacy. Surgical treatment of the injured limbs is described, as is the combined application-adsorption method of local detoxifying therapy of suppurative wounds, which, combined with staged necrosectomy, helped cope with purulent complications, prepare the wounds of the limbs to repair plastic surgery within 35 to 55 days after the injury, and rule out lethal outcomes. PMID- 7486189 TI - [Main principles of the treatment of patients during acute period of burns]. PMID- 7486190 TI - [Significance of tissue perfusion disorders and disseminated intravascular blood coagulation in the pathogenesis of post-resuscitation disease in patients with massive hemorrhage and severe multiple trauma]. AB - A total of 198 patients with massive blood loss and grave combined injury treated at the intensive care ward were monitored for 5 days. This paper presents data on the blood content of peripheral tissues assessed by rheovasography and the time course of coagulating characteristics of the blood and fibrinolysis. Despite the stabilization of arterial pressure in the first hours of treatment, disorders of tissue perfusion persisted for 5 days. Signs of disseminated intravascular coagulation were detected as soon as after 6 h in the presence of expressed progressing thrombinemia, decelerated fibrinolysis in the blood plasma, thrombocytopenia, and consumption of some blood clotting factors. According to our data, disseminated intravascular blood coagulation in the microvessels is possible after 12 h and reaches its maximum on day 2 of treatment; it persisted over the entire follow-up (5 days) period and was associated with clinically manifest involvement of the functions of parenchymatous organs and systems starting from the end of the first or beginning of the second day of treatment. PMID- 7486191 TI - [Diagnosis and treatment of DIC syndrome in patients with severe craniocerebral trauma]. AB - The hemostasis parameters are compared in 32 patients with slight (14-15 Glasgow score) and 114 with grave craniocerebral injury (3 to 7 Glasgow score) on days 1, 3, 5, and 7 after the injury. Assessment of hemostasiograms revealed a regular development of disseminated intravascular blood coagulation (DIC) in patients with grave craniocerebral injury, whereas in patients with slight injury the changes in the hemostasis system were compensatory. Early (starting from day 1) addition of anticoagulants, disaggregants, and fresh-frozen plasma to treatment protocols are advisable for patients with grave craniocerebral injuries. PMID- 7486193 TI - [Complications of chemical burns of the esophagus during combined therapy]. PMID- 7486192 TI - [Study of the mechanisms of antihypotensive action of antishock trousers in the injured with traumatic shock]. PMID- 7486194 TI - [Criteria for the choice of the extracorporeal detoxication method in patients with postoperative renal failure]. AB - Criteria for the choice of a method for extracorporeal detoxication (acetate hemodialysis, intermittent or continuous hemofiltration or hemodiafiltration, or plasmapheresis) were defined on the basis of a detailed examination of cardiorespiratory function (central hemodynamics, oxygen-transporting function of the blood) in 88 patients with acute postoperative renal failure (PRF). Multiple organ failure occurred in 90% of the patients examined in the postoperative period. The severity of visceral and metabolic disorders was the principal criterion in the choice of extracorporeal detoxication method. Hemofiltration is the method of choice for the treatment of PRF combined with multiple organ disorders, primarily with acute circulatory, respiratory, and metabolic disorders, due to its stabilizing effect on the hemodynamics and a wide spectrum of pathologic substances removed by it. Acetate hemodialysis is indicated for patients with PRF and slow recovery of renal function only after elimination of grave hemodynamic and respiratory disorders, provided there are no general cerebral symptoms, because of its negative effect on the circulation and oxygen balance of the organism and central nervous system. Plasmapheresis is a pathogenetically valid method for the treatment of the initial stages of PRF in cases with massive intravascular hemolysis and sepsis, which may be combined with other methods for extracorporeal detoxication, if necessary. PMID- 7486196 TI - [Acid-soluble blood glycoproteins in patients with diffuse purulent peritonitis]. AB - The levels of acid-soluble blood plasma glycoproteins (ABP) were compared in patients with diffuse purulent peritonitis and normal subjects. The content of protein and total carbohydrates in the ABP fraction of normal subjects was 0.68 +/- 0.08 and 0.11 +/- 0.02 mg/ml, respectively, with the carbohydrate to protein ratio 0.21 +/- 0.03. In the patients these parameters were, respectively, 1.37 +/ 0.19, 1.09 +/- 0.12, and 0.88 +/- 0.10. Possible mechanisms of accumulation of carbohydrate-enriching ABP in peritonitis patients is discussed. The carbohydrate to protein ratio is proposed to be used for assessment of the status of peritonitis patients and for prediction of the disease course and outcome. PMID- 7486198 TI - [Pulmonary ventilation with assisted pressure. Technical aspects and features of use]. PMID- 7486195 TI - [Central and cerebral hemodynamics during general anesthesia in intracranial interventions in a sitting position]. AB - Cerebral bloodflow and central hemodynamics were studied during general anesthesia in patients with tumors of the posterior cranial fossa operated on in a sitting posture. A sitting posture during general anesthesia is conductive to a marked reduction of the stroke volume and linear velocity of the cerebral bloodflow, this creating a risk of cerebral tissue ischemia under conditions of intracranial hypertension and a hardly adequate short-term compensatory increase of the total vascular resistance and "paradoxical" increase of cerebrovascular tone. Since the neurogenous mechanisms of cerebral bloodflow autoregulation are disordered during anesthesia, one should admit that stabilization of central hemodynamics at all stages of anesthesia is a most important condition for optimizing brain tissue perfusion. PMID- 7486197 TI - [Myoglobin concentration in blood: a criterion in the evaluation of muscular tissue injury in patients with prolonged crush syndrome]. AB - Blood serum levels of myoglobin, creatinine, urea, beta 2-microglobulin were measured in 37 patients with the prolonged crush syndrome (PCS) injured during the Yerevan earthquake. A drastic increase of myoglobin level in the blood was observed in all the patients by the moment of hospitalization, this level being the higher, the more severe the injury. If PCS complications developed, such as sepsis or acute renal failure, myoglobin level increased by several tens of times. Effective treatment normalized myoglobin level before discharge from hospital. PMID- 7486199 TI - [Comparative assessment of the hormonal status during N2O and xenon anesthesia using similar methods]. AB - The hormonal status of 38 surgical patients during anesthesia with nitrous oxide and Xe administered by similar methods showed that in contrast to anesthesia with N2O, Xe anesthesia helped not only preserve the adaptive potential, but even increase the defense potential of the organism during surgery. Xe anesthesia was associated with a 1.5 times less requirement in fentanyl, with the levels of stress hormones being better; Xe is the best alternative to nitrous oxide. PMID- 7486200 TI - [Spinal-epidural anesthesia: hemodynamics and hemostasis]. AB - Combined spinal epidural anesthesia using an original Portex kit was carried out in 62 patients operated on for orthopaedic, urological, and gynecological disorders. Hemodynamics was examined by tetrapolar chest rheography, hemostasis by electrocoagulography. Satisfactory blocking was attained in 98% cases, in 1 patient an accidental puncture of the subarachnoidal space with Tuohy needle occurred. Development of spinal block was associated with a reduction of heart productivity and hypocoagulation changes in the hemostasis. The parameters examined remained within the normal range of values. PMID- 7486201 TI - [Advantages of epidural anesthesia in patients with end-stage chronic renal failure]. AB - The authors analyze the experience gained in anesthesiological management of 667 surgeries in patients with the end-stage chronic renal failure. 206 patients were operated on under epidural anesthesia and 461 under general anesthesia. The technique of anesthesia, preparation of patients, and the management during and after surgery are described. 63 hemodialysis procedures were performed for 4 h before and 154 for 12 h after surgery. The complications occurring during and after anesthesia by both methods are analyzed. Epidural anesthesia was found to be more safe for patients with the end-stage chronic renal failure. General anesthesia more often led to hemodynamic, respiratory, metabolic disorders, and other hemostasis disturbances. PMID- 7486202 TI - [Use of antibiotic thienam in critical care patients]. PMID- 7486203 TI - [Cerebral oximeter INVOS-3100]. AB - A new device is described: cerebral oximeter intended for monitoring cerebral oxygenation and early diagnosis of cerebral ischemia and hypoxia. The advantages and shortcomings of the proposed method are analyzed in comparison with other current methods for the early diagnosis of cerebral ischemia. PMID- 7486204 TI - [The system of readiness to take care of medical consequences in disaster situations]. AB - The main task of regional catastrophe medicine service in liquidation of medical consequences of emergency situations is the immediate adequate reaction which is aimed at minimizing the sanitary loss. This task can be effectively solved only on condition of permanent improvement of interaction between the managing and managed systems of public health and coordination of their actions with those of nonmedical services. An important role is played by the actualized (constantly renewed) data bases "Potentially Hazardous Objects", "Regional Infrastructure", "Medicosanitary Means", etc., which help simulate various emergency situations with due consideration for the season, time of the day, and socioeconomic features of a region in order to create variants of effective work of all services engaged in the liquidation of medical consequences. PMID- 7486205 TI - [Hemodialysis with electrochemical regeneration of dialysis solution]. AB - The method of electrochemical regeneration of dialysis solution is virtually as effective as the traditional hemoperfusion system in removing creatinine and potassium ions from the body, but inferior to it as regards urea and inorganic phosphorus and superior in removal of medium-weight toxins. A more smooth reduction of urea level in the blood provides relatively stable hemodynamics in patients liable to hypotension. Electrochemical regeneration permits a continuous purification of dialysis solution, the regeneration block may be used many times, and sodium hypochlorite produced during electrolysis operation maintains the sterility of the system. Electrochemical regeneration does not require water purification systems, water communication, and dialysis concentrate, this permitting the performance of detoxication in rooms without special equipment. PMID- 7486206 TI - [Experience in the treatment of patients with prolonged crush syndrome]. PMID- 7486207 TI - Comparative study of left ventricular function in patients with unstable angina, non-Q wave myocardial infarction and stable angina pectoris: assessment with atrial pacing and digital ventriculography. AB - To compare left ventricular global and segmental function at rest and during right atrial pacing in patients with unstable angina, non-Q wave myocardial infarction, and stable angina (class III angina), low-dose digital subtraction ventriculography was performed at rest and after abrupt cessation of pacing in 42 patients with unstable angina, 8 patients with non-Q wave myocardial infarction and 15 patients with stable angina during selective coronary arteriography. Left ventricular ejection fraction was significantly lower at rest in patients with unstable angina (P < 0.01) and non-Q wave myocardial infarction (P < 0.05) and during pacing (P < 0.01). These two groups of patients had significantly higher values of left ventricular end-diastolic and end-systolic volumes at rest and during pacing as compared with stable angina group. In comparing various clinical patterns of unstable angina, ejection fraction was significantly (P < 0.05) lower during pacing in patients with crescendo angina than in new-onset angina. However, ejection fraction was significantly (P < 0.01) lower in crescendo angina only at rest as compared with rest angina. The length of zone of severe hypokinesia was greater in unstable angina (P < 0.01) as well as in non-Q wave myocardial infarction (P < 0.05) both at rest and during pacing as compared with stable angina. Contractility of region of hypokinesia during pacing was higher (P < 0.01) in stable angina than in unstable angina and non-Q wave myocardial infarction. In analyzing segmental function in various subgroups of unstable angina, the authors found that the length of total hypokinesia was significantly higher (P < 0.05) during pacing in crescendo angina than in new-onset angina. Contractility of region of hypokinesia was lowest at rest and during pacing in patients with crescendo angina. This study demonstrates that patients with unstable angina as well as non-Q wave myocardial infarction were characterized by more pronounced global and segmental left ventricular dysfunction at rest and during pacing as compared with patients with stable angina, which may explain the poorer prognosis in the former two groups. This study also shows that patients with crescendo angina have more profound left ventricular global and regional dysfunction as compared with patients with new-onset as well as rest angina. PMID- 7486208 TI - Color Doppler imaging of basal cerebral arteries: normal reference values and clinical applications. AB - Transcranial color-coded duplex ultrasonography is a new diagnostic procedure allowing the visualization of major intracranial vessels. The purpose of this report is to describe this method and to evaluate its practical potential in the routine examination of basal cerebral arteries. The results from the examination of 96 patients are presented. By means of color coding of Doppler frequencies, the major intracranial vessels were identified by nature of their anatomic location with respect to the echogenic brainstem structures in all subjects. The average maximal systolic velocity was, in the middle cerebral artery, 108 +/- 15 cm/s; in the anterior cerebral artery, 96 +/- 15; in the posterior cerebral artery, 76 +/- 16; and in the basilar artery, 59 +/- 17 cm/s. The determination of the angle between the ultrasonic beam and the examined vessel improved the accuracy of flow velocity measurements in comparison with conventional transcranial Doppler ultrasonography. The exact measurement of blood flow velocity in several segments of the visualized vessel proved helpful in the assessment of pathologic findings, especially in differentiation between a stenosis of the intracranial vessel and a vasospasm. PMID- 7486209 TI - Diagnostic value of the arithmetic sum of the ST segment of inferior and V2 leads, II + V2, III + V2 and aVF + V2 in identifying the artery responsible for inferior acute myocardial infarction. AB - In order to evaluate electrocardiographic changes in the diagnosis of the artery responsible for inferior myocardial infarction, a prospective study was performed on inferior and V2 ST segment deviation and its correlation using the arithmetic sum: II + V2, III + V2, and aVF + V2. A group of 66 patients with inferior acute myocardial infarction (AMI) was studied. A standard 12-leads electrocardiogram was performed within six hours of the onset of chest pain. Coronary arteriography was performed on each of the patients between one and twelve weeks after infarction. Right coronary artery (RCA) lesion was found in 46 patients, 27 at a proximal level and 19 at a distal level; in 20 patients the left circumflex coronary artery was affected. The isolated value of the magnitude of the inferior ST segment is not an efficient parameter for identifying the artery responsible for inferior AMI. In lead V2 all the patients with a lesion of the left circumflex artery showed ST segment depression > or = 1 mm (P < 0.001) and all those presenting ST segment elevation had stenosis of the proximal RCA. The most useful parameters for identifying the artery responsible for inferior AMI, with 100% specificity are: (1) for occlusion of the RCA, the arithmetic sum of ST segments: aVF + V2 > 0, with 86.9% sensitivity (P < 0.001); (2) for occlusion of the left circumflex artery III + V2 < 0, with 90% sensitivity (P < 0.001); and (3) for proximal occlusion of the RCA: aVF + V2 > or = 1, with 96.2% sensitivity (P < 0.001). No specific marker was observed for distal occlusion of the RCA. The value of the arithmetic sum of the ST segment: III + V2 between 0 and 0.9 was the most significant, with 94.7% sensitivity and 95.7% specificity (P < 0.001). PMID- 7486210 TI - Effect of methylprednisolone on metabolism and contractility in the stunned myocardium. AB - The effect of glucocorticoid on the metabolism and contractility in the stunned myocardium was examined by phosphorus 31 nuclear magnetic resonance (31P-NMR) in Langendorff rabbit hearts by use of an artificial blood substitute, perfluorochemical emulsion Flusol-43. After normothermic global ischemia of fifteen minutes, postischemic reperfusion of sixty-five minutes was carried out. Methylprednisolone sodium succinate (MPSS) was administered either prior to global ischemia or during postischemic reperfusion. Adenosine triphosphate (ATP), creatine phosphate (CrP), inorganic phosphate (Pi), pH, left ventricular systolic developed pressure (LV DevP) and coronary flow were continuously measured. Thirty six hearts were divided into three experimental groups consisting of 12 hearts each; CONT consisted of controls, Pre-MPSS perfusion with MPSS-containing solution (10(-4)M) from forty-five minutes prior to global ischemia, and Post MPSS with the same MPSS solution immediately after postischemic reperfusion. Pre MPSS showed a significant inhibition of the increase in Pi and of the decrease in ATP and pH during global ischemia, in comparison with the other groups, and a suppression of the overshoot of CrP observed immediately after postischemic reperfusion. LV DevP of Pre-MPSS showed a marked improvement during the postischemic reperfusion as compared with CONT. In Post-MPSS, Pi was significantly increased and ATP decreased during the postischemic reperfusion as compared with the other two groups. There were no differences in coronary flow during postischemic reperfusion among the three groups. In conclusion MPSS has a beneficial effect on metabolism and contractility of the stunned myocardium when it is administered prior to ischemia. PMID- 7486213 TI - The effect of streptokinase on return to work 18 months after a first myocardial infarction. AB - In order to assess the impact of thrombolytic therapy on return to work eighteen months after a first myocardial infarction, 32 patients treated with streptokinase were compared with 30 patients not treated with streptokinase. The study was designed as a historical cohort study. The patients in both groups had continuous chest pain of less than six hours prior to admission and electrocardiographic changes consistent with acute myocardial infarction (MI). The two groups were comparable with respect to medical variables related to their myocardial infarction and to educational level. A total of 17 patients (53%) in the streptokinase group and 16 (53%) among controls had stopped working eighteen months after their MI. An association between the treatment and the working status could not be found (relative risk = 1.0) nor could it be found if the figures were corrected for deaths and retirements because of age. In conclusion, this study could not demonstrate any beneficial effect of thrombolytic therapy on the return to work eighteen months after an initial myocardial infarction. PMID- 7486212 TI - Urine C-peptide and atherogenic risk factors in diabetes mellitus: relevance to "syndrome X". AB - The relation between the C-peptide concentration in twenty-four-hour urine specimens and atherogenic risk factors was investigated in 38 patients with noninsulin-dependent diabetes mellitus in an attempt to determine the significance of urine C-peptide in diagnosing "syndrome X," which is characterized by insulin resistance. Weak positive correlations between twenty four-hour urine C-peptide concentration and body mass index, systolic blood pressure (BP), diastolic BP, serum total cholesterol, and serum triglyceride were detected. A weak negative correlation was also apparent between urine C-peptide and serum high-density lipoprotein (HDL). The body mass index and serum triglyceride of patients with urine C-peptide excretion of > 100 micrograms/day were significantly higher than those in patients with normal urine C-peptide excretion (< 100 micrograms/day) (P < 0.01 and P < 0.02, respectively). Systolic BP, diastolic BP, serum total cholesterol, and serum HDL did not differ significantly between the two groups of patients. Results indicate that twenty four-hour urine C-peptide concentration is of significance in determining whether a patient has a tendency to insulin resistance but has only limited value as a quantitative measure of endogenous insulin secretion. PMID- 7486214 TI - Normal-appearing captopril MAG-3 renal scintigraphy in hemodynamically significant renal artery stenosis. A case report. AB - The finding from a normal-appearing angiotensin converting enzyme (ACE)-inhibitor renal scan is generally reassuring to the physician screening for renovascular hypertension. In fact, the false-negative rate for captopril scintigraphy is very low. Possible reasons for false-negative scans have not been well documented. A fifty-two-year-old man was evaluated and found to have renovascular hypertension on two occasions, at initial presentation and again eight months later (restenosis had occurred). Renovascular hypertension was present on both occasions as judged by decline of blood pressure following angioplasty of right renal artery stenosis (approximately 80% and approximately 70% stenosis on the two occasions, respectively). However, ACE-inhibitor renal scanning with 99mTc MAG-3 gave disparate results on the two occasions. The first study using oral captopril (25 mg) indicated a low probability of renal artery stenosis, whereas the second study done with the patient regularly taking lisinopril (10 mg daily) was markedly positive. Possible reasons for the initial negative study include poor absorption of oral captopril or inadequate inhibition of the renin angiotensin system by the 25 mg dose. PMID- 7486211 TI - Endothelium-derived vasoactive substances in Bartter's syndrome. AB - An imbalance between endothelium-derived vasoactive substances such as endothelin and endothelium-derived nitric oxide (NO) might be viewed as a possible determinant of vascular hyporeactivity. To check this possibility the authors evaluated the role of endothelin and NO in the reduced vascular reactivity of Bartter's syndrome. Plasma immunoreactive endothelin (22.07 +/- 7.06 vs 13.80 +/- 1.43 pmol/L, P < 0.011), urinary excretion of NO2- (0.28 +/- 0.10 vs 0.15 +/- 0.02, mumol/mumol of urinary creatinine, P < 0.01) and NO3- (0.17 +/- 0.07 vs 0.09 +/- 0.02 mumol/mumol of urinary creatinine, P < 0.011), and forearm resting blood flow (FRBF) (6.67 +/- 1.69 vs 4.30 +/- 0.38 mL/m'/100 mL, P < 0.005) were increased in patients with Bartter's syndrome in comparison with normal controls (C). No difference in postischemic maximal FBF was found (34.14 +/- 4.67 vs 31.35 +/- 2.86 mL/minute/100 mL), while patients showed a slower recovery after peak flow (PF) (77.57 +/- 61.35 vs 9.42 +/- 3.69 seconds, P < 0.013). Higher plasma endothelin supports the defect in vascular reactivity of Bartter's syndrome already shown for angiotensin II and norepinephrine and is in keeping with the altered intracellular calcium signaling previously demonstrated by the authors in this syndrome. The increased excretion of NO2- and NO3- in this syndrome, together with the higher FRBF and the slower recovery of the FBF and PF, argues in favor of an increased NO synthesis in Bartter's syndrome and of assigning it a role in the vascular hyporeactivity of Bartter's syndrome. PMID- 7486215 TI - Transient cortical blindness following bypass graft angiography. A case report. AB - Transient cortical blindness, an uncommonly recognized complication of cerebral angiography, is an exceedingly rare event after cardiac catheterization and angiography. This report describes a sixty-two-year-old patient who had transient cortical blindness following bypass graft angiography. In this case, the authors showed that cortical blindness was associated with the breakage of the blood brain barrier (BBB) and an increase in vascular permeability rather than with primary cerebral circulatory insufficiency. When the possibility exists that an excess volume of contrast medium may enter the cerebral circulation as in this case, that is, following a coronary artery bypass graft (CABG) using the internal mammary artery (IMA), precautionary measures may be necessary such as changing the type of contrast medium to be used or decreasing the volume injected. When cortical blindness occurs, it is a serious clinical problem whether transient or permanent. Therefore, the circumstances leading to this complication should be understood to determine suitable treatment and management. PMID- 7486216 TI - Unusual coronary collateral circulation: filling of a totally occluded left anterior descending artery by direct continuity from a left posterior descending artery. A case report. AB - A patient with an unusual coronary collateral circulation is presented. The left anterior descending artery, which was totally occluded proximally, was filled directly by a continuation of the left posterior descending artery. A thallium stress test revealed normal perfusion in the territory of the left anterior descending artery. PMID- 7486218 TI - Anomalous origin of left circumflex artery in a scimitar syndrome. A case report. AB - This report concerns a twenty-seven-year-old woman with scimitar syndrome of abnormal origin of the left circumflex artery from the pulmonary artery. Operation was performed to redirect the abnormal venous return to the left atrium and abnormal origin of the left circumflex artery to the aorta. Unfortunately, pulmonary hypertension with hemoptysis occurred nineteen months after the operation. A second angiogram showed total occlusion of the graft to the left circumflex artery and impaired left ventricle function, rather than thrombosis of the anastomosis to the scimitar vein, as the cause of pulmonary hypertension. PMID- 7486217 TI - Acute myocardial infarction in a patient with primary coronary dissection and severe coronary vasospasm. A case report. AB - A case of myocardial infarction associated with coronary artery dissection in a thirty-five-year-old woman is reported. An emergent coronary angiogram revealed extensive dissection and thrombosis in the right coronary artery; in addition, severe vasospasm was observed in the left coronary artery on the next day. She was successfully treated with intracoronary thrombolysis and intra-aortic balloon pumping. The patient is alive and well two years after infarction. PMID- 7486220 TI - Pulmonary arteriovenous malformation. PMID- 7486219 TI - Severe acute hand ischemia following an accidental intraarterial drug injection, successfully treated with thrombolysis and intraarterial Iloprost infusion. Case report. AB - A young woman with acute upper extremity edema and ischemia after intraarterial drug injection is presented. Unsatisfactory results from standard treatment were the reason for changing the therapy to temporary thrombolysis, which led to significant improvement. Some days later severe impairment forced another attempt at applying standard therapy, again unsuccessful. Only after continuous intraarterial infusion of Iloprost, a new improvement occurred and saving of the hand was possible. It became obvious that more effective therapeutic measures ought to be applied when severe hand ischemia following intraarterial drug injection is present. PMID- 7486221 TI - Myocardial scintigraphy with 99mTc-sestamibi in children with Kawasaki disease. AB - Kawasaki disease (KD) is a systemic vasculitis syndrome of early childhood. It involves particularly the coronary arteries and may cause aneurysms and thrombotic occlusions. Echocardiography is the most useful method of detecting coronary aneurysms. Nevertheless, obstructive lesions are difficult to evaluate and often need invasive coronary angiography. An important feature of this disease is the possibility of finding coronary pathology several years after the onset. This characteristic makes KD an important cause of coronary artery disease (CAD) in young adults. Thus patients with KD and previously diseased coronary arteries should be kept under long-term control. However, coronary angiography is invasive and cannot be performed repeatedly, especially in young patients. As an alternative, thallium 201 scintigraphy has been employed, but its low-energy photons are suboptimal for standard gamma-camera imaging, particularly in children aged less than three years. To verify the usefulness of a noninvasive assessment of myocardial perfusion, the authors used rest and dipyridamole 99mTc Sestamibi scan in 15 children (ranging from one to six years of age) with Kawasaki's cardiac involvement. Coronary aneurysms have been demonstrated by echocardiography in 12 patients; 8 patients were also submitted to cardiac catheterization. The sensitivity of 99mTc-Sestamibi imaging for detection of overall coronary lesions was 88% and the specificity was 93%. These data suggest that rest/dipyridamole 99mTc-Sestamibi scintigraphy is an accurate and noninvasive method for the detection and follow-up of Kawasaki's cardiac damage even in patients aged one year. PMID- 7486223 TI - Angioplasty-induced pseudoaneurysm successfully treated with a Palmaz stent. A case report. AB - Pseudoaneurysm formation and vessel rupture are considered rare complications of percutaneous transluminal angioplasty (PTA), and few reported cases exist in the literature. The true incidence of pseudoaneurysm formation is unknown, for follow up angiography is not routinely performed unless symptoms recur or persist. Currently, the only intervention widely accepted for angioplasty-induced pseudoaneurysm involves surgical excision or repair. The authors describe a case of successfully treated pseudoaneurysm with a noncovered stent. PMID- 7486222 TI - Increased lung uptake during myocardial scintigraphy improves the detection and localization of coronary artery disease. AB - Increased lung uptake during exercise thallium 201 scanning denotes severe ventricular hypofunction. To assess whether there is any relationship between this finding and the stenosis of a single coronary artery, the authors studied 140 patients (age 57.5 +/- 12 years) with both exercise thallium and coronary angiography within three months. A planar view was acquired for two hundred forty seconds immediately postexercise and prior to the single photon emission computed tomography acquisition. Two identical regions of interest outlining the left ventricle and the right lung in the planar view were used for calculation of the lung uptake, expressed as a lung-to-heart counts ratio. The patients were divided into four angiographic groups: (I) lesions at the left anterior descending (LAD), n = 35; (II) stenosis at the circumflex (CRX), n = 35; (III) stenosis at the right coronary artery (RCA), n = 35; and (IV) another group of 35 normal subjects serving as controls. No patient had a previous myocardial infraction (MI) or any cardiac operation. Patients in groups I (0.53 +/- 0.094) and II (0.44 +/- 0.072) had higher lung uptake than those in groups III (0.43 +/- 0.061) or IV (0.42 +/- 0.050). There was no difference between groups III and IV, or II and IV, but the difference between I and IV was of statistical significance (P < 0.001).(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7486224 TI - Improved skin blood flow and cutaneous temperature in the foot of a patient with arteriosclerosis obliterans by vasopressin V1 antagonist (OPC21268). A case report. AB - A seventy-four-year-old woman with arteriosclerosis obliterans, diabetes mellitus, and hypertension was admitted for the treatment of intermittent claudication and coldness and pain in the right lower extremity. After the administration of a vasopressin V1 antagonist, OPC21268, the symptoms were markedly improved. Furthermore, blood flow in the dorsalis pedis artery and the cutaneous temperature in the right foot increased in response to acute and chronic administration of OPC21268. OPC21268 may be a new useful therapeutic tool for the treatment of arteriosclerosis obliterans. PMID- 7486225 TI - Diagnosis of lymphangitic carcinoma to lung by sputum cytology: case report. AB - Documenting lymphangitic metastases to the lungs may be therapeutically important. Several techniques have been described to this end. Sputum cytology, the least invasive and expensive, is the most available. It has a role that may be overlooked but that should be emphasized, since abundant malignant cells in sputum may be associated with this pattern of tumor spread. This case is the first, to the authors' knowledge, in which the pathologic findings associated with malignant cells in lymphangitic carcinomatosis were demonstrated in sputum in the absence of tumor mass invasion of large airways. PMID- 7486227 TI - Portal vein thrombosis due to antithrombin III deficiency. A case report. AB - The authors report a case of intrahepatic portal vein thrombosis in a sixty-four year-old Japanese man with antithrombin III deficiency who successfully underwent cholecystectomy and common bile duct exploration. To their knowledge, this is the first report of portal vein thrombosis due to primary antithrombin III deficiency. PMID- 7486229 TI - Flow pattern of four pulmonary veins in a case with prolapsing left atrial myxoma. A case report. AB - The authors report the flow pattern of the four pulmonary veins in a case with prolapsing left atrial myxoma. The early systolic flow was reversed, midsystolic forward flow was diminished, and diastolic forward flow was augmented and decelerated rapidly in all four pulmonary veins. This phenomenon is attributed to the dynamic movement of the myxoma. PMID- 7486226 TI - F.P. Weber syndrome associated with a brachial artery aneurysm. A case report. AB - Klippel-Trenaunay syndrome is characterized by the triad of unilateral port-wine hemangiomas, varicose veins, and hypertrophy of bone and soft tissue affecting one or more limbs. The rare F.P. Weber syndrome describes the mentioned entity and additional arteriovenous malformations. The association of an arterial aneurysm with the F.P. Weber syndrome has never been described in the current literature. A case of a brachial artery aneurysm in a patient with F.P. Weber syndrome is presented and the etiology of arterial aneurysm combined with congenital vascular abnormalities is discussed. PMID- 7486228 TI - Drug injection in jugular veins: a new risk factor for vascular diseases in HIV infected patients? A case report. AB - The authors describe a rare case of diffused thrombosis of the superficial veins in the whole body and periphlebitis with perivascular abscesses in an human immunodeficiency virus (HIV)-infected drug abuser who was using neck veins to inject cocaine and heroin. In addition the patient presented oral candidiasis, hepatitis C virus infection, bronchopneumonitis, and endocarditis of the tricuspid valve with valvular failure. The conditions of the patient needed repeated vascular catheterizations for therapy administration. Similar pathologies, in HIV-infected patients, highly increase the risk of opportunistic infections, especially in the encephalic territory; in addition the need for vascular catheterizations represents a further risk factor for bacterial infections. PMID- 7486231 TI - Diagnostic dilemma of an intramedullary vascular malformation. A case report. AB - A young man was seen at a neurologic center for a slowly progressive myelopathy involving the lower limbs in 1983. After examination and investigations, a diagnosis of chronic tuberculous spinal arachnoiditis was made and the patient was administered antituberculous treatment. However, his condition continued to deteriorate. He was reviewed at another center in 1985, where a diagnosis of demyelinating disorder was made, for which he was treated. However, this also did not help, and he was diagnosed as having a nonspecific spinal arachnoiditis at yet another center in 1988. His condition continued to worsen, and after ten years of symptoms, when magnetic resonance imaging (MRI) was done, an intramedullary cystic lesion opposite D-12 vertebrae was seen, which on histopathology was proved to be a vascular malformation. After excision of the lesion his neurologic status has been slowly improving. The conditions that a malformation could mimic, misleading a clinician, and the role of MRI in clinching the diagnosis are highlighted. PMID- 7486230 TI - Four-barrel aortic dissection involving the ascending aorta. A case report. AB - A fifty-four-year-old woman was evaluated at the authors' hospital for symptoms of repeated chest and back pain. The symptoms had recurred for five months prior to admission. A dynamic computed tomographic scan was performed on admission and demonstrated an enlarged ascending aorta with four barrels secondary to split intimal flaps. This case is an extremely rare example of a patient with a four barrel aortic dissection involving the ascending aorta. PMID- 7486232 TI - Migration behavior of human smooth muscle cells cultivated from restenotic and primary lesions. AB - Subintimal smooth muscle cell (SMC) migration is considered an essential determinant of arteriosclerosis and neointimal formation. In this study, a cell culture model was established to characterize migration activity of SMCs originating from restenotic and primary lesions. Plaques from symptomatic stenoses of 32 patients (19 men, 13 women; 4 carotid, 17 peripheral, 11 coronary lesions) were removed by percutaneous atherectomy or direct operative approach. Ten patients suffered from recurrent stenosis. Cell cultures were established by explantation of tissue samples. By indirect immunofluorescence microscopy, SMCs were shown to be the predominant cell type of all advanced lesions irrespective of their origin. The spontaneous cellular motility of SMCs was analyzed in vitro by means of a computer-assisted observation system. Cells of all groups exhibited random motility. SMC migratory velocity was found to be significantly (P < 0.001) greater in cells from restenotic lesions than in those from primary plaques. In conclusion, migration behavior of human SMCs originating from arteriosclerotic lesions may be quantified in vitro as a functional determinant characterizing restenotic versus primary lesions. PMID- 7486233 TI - Reduction of left ventricular hypertrophy with St. Jude Medical 19 mm valve prosthesis. AB - To determine the permissible range of the St. Jude Medical 19 mm valve (SJM19) as an aortic prosthesis, transvalvular maximal pressure gradient (maxPG) and postoperative reduction of left ventricular (LV) hypertrophy were examined in 21 patients (body surface area [BSA] 1.28-1.56 m2, mean 1.42 m2). Doppler echocardiographically measured maxPG at rest was 31 +/- 16 mmHg, which showed no significant difference from the differences in patients' diseases (aortic regurgitation = AR or stenosis = AS), or BSA. The maxPG, also measured during dobutamine administration when they were less than 40 mmHg at rest, increased significantly from 21 +/- 11 mmHg to 32.1 +/- 15 mmHg, having no differences from the differences in diseases or BSA. Reduction of LV hypertrophy was achieved postoperatively (mean postoperative period of ten months), with the reduction rates of 26 +/- 19% in LV mass and 21 +/- 15% in LV cross-sectional area. No significant differences were found between the reduction rates and the differences in the patients' diseases or BSA. These results indicate that, following the implantation of SJM19, LV hypertrophy was reduced in patients with AR as well as with AS, when their BSA values were less than 1.6 m2. However, limited use is recommended because the increase in pressure gradient during exercise cannot be ignored. PMID- 7486234 TI - Electrocardiographic diagnosis of the responsible coronary artery in acute inferior myocardial infarction through right chest leads V3R-V8R. A prospective study. AB - In order to identify the electrocardiographic changes that occur in right-chest leads V3R-V8R for the most significant diagnosis of the responsible coronary artery of acute myocardial infarction, the authors performed a prospective study on 66 patients in whom coronary arteriography was done between the first and twelfth weeks after suffering the infarction. Electrocardiograms were done within the first six hours after the onset of symptoms. Lesions of the right coronary artery were found in 46 patients--27 at a proximal level and 19 at a distal one- and in 20 patients the circumflex coronary artery was injured. The electrocardiographic findings were studied in 2 groups of leads: V3R-V4R and V5R V8R. An ST elevation equal to or higher than 0.5 mm and the presence of Q waves in V3R-V4R are specific markers of lesions of the right coronary artery (P < 0.001). Lowering of the ST segment in V3R-V4R is a specific marker of a circumflex artery lesion (P < 0.001). An ST elevation equal to or higher than 1 mm in V3R-V4R is specific for a proximal lesion of the right coronary artery (P < 0.001). No specific marker for a lesion of the distal right coronary artery was identified, its more significant characteristic being an "isoelectrical" ST segment (between 0 and 0.4 mm), an rS morphology and positive T waves in V3R-V4R. PMID- 7486235 TI - A randomized trial of E5510 versus aspirin in patients with transient ischemic attacks. The Japanese E5510 TIA study-1 (JETS-1) Group. AB - In a randomized double-blind trial, the Study Group compared the efficacy of E5510, a novel antiplatelet agent, and aspirin in preventing the recurrence of transient ischemic attacks (TIA). In total, 227 patients who suffered from TIA in the twelve weeks prior to the study were enrolled. They were randomly allocated to three treatment groups, ie, 71 patients in the E5510 4 mg group, 77 patients in the E5510 2 mg group, and 79 patients in the aspirin 324 mg group, and were treated for twelve to twenty-four weeks. The incidence of recurrent TIA or stroke was 21.5% in the aspirin group and was significantly lower in the E5510 groups, being 8.5% in the 4 mg group (P < 0.05) and 11.7% in the 2 mg group (P < 0.05). Adverse events were observed in 5 cases in the 4 mg group, in 8 cases in the 2 mg group, and in 10 cases in the aspirin group, but none of them were serious. Since safety was judged to be comparable among the three groups, E5510 appears to be an antiplatelet agent for the treatment of TIA with a clinical benefit over aspirin. PMID- 7486236 TI - The need for video imaging. PMID- 7486237 TI - Impacted canines. PMID- 7486239 TI - Communication in orthodontic treatment planning: bioethical and informed consent issues. AB - Orthodontic treatment planning is an interactive process in which the patient or parent and the orthodontist serve as co-decision makers. As in most partnerships, there is a natural tension between the orthodontist and the patient because of differences in their frames of reference. The orthodontist generally is influenced more by the objective findings (the problem list), whereas patients are guided more by subjective issues related to their perceived needs, desires, and values. The art of careful probing and listening to the patient as part of the treatment planning process is an essential skill. One of the most difficult situations in contemporary orthodontics is presented by the patient with a jaw discrepancy for which the alternative treatments are orthodontic camouflage through dental compensation or surgical-orthodontic correction. Computer imaging to simulate the probable treatment outcomes can facilitate communication about these alternatives by eliminating misconceptions. Full disclosure and the consideration of all viable treatment alternatives have great benefits from a risk management standpoint, in addition to their bioethical merits. PMID- 7486238 TI - Case report: treatment of dental asymmetry. AB - The treatment of dental asymmetries often comprises several treatment alternatives. This case report describes the treatment alternatives for an asymmetry generated secondary to surgical removal of an odontoma that included the germ of the lower left lateral incisor. The opening of the space was chosen based on the patient's wish. The asymmetrical biomechanical force system used for the correction of the midline is presented as free-body diagram. PMID- 7486240 TI - The influence of video imaging on patients' perceptions and expectations. AB - Seventy-four patients, ranging in age from 15 to 50 and presenting with a moderate to severe dentofacial deformity requiring orthognathic surgery, were randomly assigned to a video imaging or a standard case presentation (CP) group. Motives for treatment did not differ significantly between the video image and standard CP groups. Two weeks after the case presentation, 60% of the standard CP group and 74% of the video image group ranked the anticipated change in facial appearance as an important factor in making a treatment decision. Seventy percent of the standard CP group and 83% of the video image group agreed with the statement that surgery would be necessary to correct their problem. In the standard CP group, 47% of the patients ranked dental casts as the most helpful physical record in making a treatment decision, and 46% said the acetate profile tracing was the most helpful tool for understanding what to expect from treatment. In the video imaging group, 42% of the patients said the video imaging presentation was the most helpful tool in making a treatment decision, and 39% of the patients said it was the most helpful tool for understanding treatment expectations. The proportion of patients in the video image case presentation group with elevated self-image expectations following treatment was significantly higher (P = .045) than in the standard case presentation group. The presentation of video images appears to be a valuable information source for conveying treatment options to patients, but caution may be needed to prevent elevated or possibly unrealistic treatment expectations. PMID- 7486241 TI - A comparison between radiographic and sonically produced cephalometric values. AB - Cephalometric radiography has become a standard and invaluable means of obtaining diagnostic information for the management of malocclusion and skeletal disharmony. However, concerns over radiographic exposure, particularly in growing individuals, may limit its use, especially in longitudinal analyses. Less invasive means of obtaining vital information would be desirable. A recently introduced system (Digigraph, Dolphin Imaging Systems; Valencia, Calif) provides sonically produced representations of cranial landmarks and has been introduced by its manufacturer as an alternative to standard cephalometric radiography. The purpose of the study was to compare the validity and reproducibility of cephalometric values generated sonically on a digital image analyzer (Digigraph) with those obtained from standard cephalometric radiographs for 43 different measurements. Although 58.1% of the sonically produced measurements showed significant correlation with radiographically produced measurements, there were no trends observed for correlation, either in dental or skeletal structure classifications. Additionally, this study found the data generated from the digital image analyzer to be markedly variable, while the radiographically obtained data were reproducible. PMID- 7486242 TI - Reliability of computer-generated prediction tracing. AB - The reliability of a commercially available computer prediction program (Quick Ceph II) was evaluated using pretreatment and posttreatment cephalograms of 30 patients who were treated during an active period of growth. The computer prediction was compared with the actual treatment result, and the growth forecast with the computer program was compared with the growth forecast using a manual method. Using paired student's t-tests, predictions for 5 of the 10 variables measured were found to be statistically reliable. Comparing the relative accuracy of growth prediction in terms of absolute values, the computer came closer to the actual result in four of the nine variables, while the manual method came closer in three variables. Predictions for the other two variables were virtually the same using both methods. The manual method of prediction was sufficient to give a reasonably good graphic representation of growth changes to create a VTO. However, the computer offers the added advantages of quicker access to information and somewhat greater accuracy in producing the tracing, as well as its use in patient education. PMID- 7486243 TI - Changes in soft tissue profile of African-Americans following extraction treatment. AB - The purpose of this investigation was to determine changes in soft tissue profile of African-Americans following orthodontic treatment involving extraction of four premolars. The sample consisted of pretreatment and postreatment lateral cephalometric radiographs of 30 males and 30 females of African-American descent exhibiting bimaxillary protrusion. The age of the patients ranged between 10 years 4 months and 17 years 6 months at the start of treatment. Average time between pretreatment and postreatment radiographs was 2 years 11 months in the male group and 3 years 3 months in the female group. Changes in the dentofacial complex and facial soft tissue as a result of treatment and growth were evaluated with cephalometric analysis. Student's t-tests were performed to compare differences. Nasolabial angle increased 9.1 degrees in males and 7.1 degrees in females. Upper lip procumbency relative to SnPg' decreased 1.5 mm in males and 1.7 mm in females. Lower lip retraction relative to SnPg' was 2.7 mm in males and 2.5 mm in females. PMID- 7486244 TI - Arch dimensional changes in children with idiopathic short stature treated with recombinant growth hormone: a five-year study. AB - Recombinant human growth hormone (rhGH) increases stature when administered to non-GH-deficient idiopathic short statured children. The aims of this investigation were to determine pretreatment arch dimensions of short statured children (height > or = 2 S.D. below mean for age) and to evaluate their response to rhGH administration by measuring arch-dimensional changes over 5 years of rhGH treatment. Dental casts of 28 short subjects (22 male, 6 female) and of age/gender-matched controls of normal stature were analyzed using a digital imaging system. Four measures of arch width and one of arch depth were calculated for each maxillary and mandibular cast. Subjects receiving orthodontic care were eliminated from the study at the initiation of treatment. Z-scores were calculated to allow for pooling of data. Prior to rhGH treatment, all arch dimensions of the short statured subjects were smaller than the controls, with the exception of mandibular arch depth. Arch dimensions of the rhGH subjects did increase with the continued administration of rhGH. The control group also showed a significant trend for the arch dimensions to increase over time. The influence of rhGH treatment on arch dimensional changes over time remains equivocal based on the results of this investigation. PMID- 7486245 TI - Number and distribution of silver-stained nucleolar organizer regions and evolutionary relationships in domestic pigs. AB - Variation in the numbers of silver-stained nucleolar organizer regions (Ag-NORS) were examined in 36 breeds of domestic pig of different geographic origins and five subspecies of wild boar. The relationship between Ag-NORs and evolution of domestic pigs was investigated. In all pigs observed, Ag-NORs were localized on the secondary constriction of chromosomes 10 and 8. The mean Ag-NOR numbers varied from 2.0-4.0, and decreased gradually with the different geographical distribution from south to north in China and from east to west in Europe. This regular change was caused mainly by the differences of frequency in chromosome 8 Ag-NOR type and was closely related to the evolution of domestic pig breeds. PMID- 7486246 TI - A panel of polymorphic bovine, ovine and caprine microsatellite markers. AB - A panel of 81 new polymorphic bovine microsatellite markers is described, together with further information on a previously reported group of 16 markers. The mean polymorphism information content of the 97 markers determined in 20 cattle was 0.66. Seventy-three of these markers have been assigned to chromosomes by either linkage analysis or use of hybrid cell panels. Thirty-nine of the markers were polymorphic in sheep, and 32 were polymorphic in goat. This study identified a set of 18 robust markers that were polymorphic in all three species and that covered 14 bovine chromosomes. This provides a single group of markers, which would be suited to genetic distance analysis and parentage control in cattle, sheep and goat. PMID- 7486247 TI - Biochemical identification of the bovine blood group M' antigen as a major histocompatibility complex class I-like molecule. AB - Absorption and elution experiments showed that it was impossible to separate antibodies against blood group factor M' from antibodies against bovine lymphocyte antigen (BoLA) A16 in an antiserum showing haemolytic activity against M' as well as lymphocytotoxic activity against BoLA-A16. To elucidate the structural relationship between BoLA-A16 and blood group antigen M', immunoprecipitation experiments on red and white cell lysates isolated from M' A16 positive and negative cattle were carried out. These results showed that M(r) 44,000 and M(r) 12000 polypeptides can be precipitated from both red and white cells isolated from M'-A16 positive animals, whereas no bands were seen in M'-A16 negative animals in precipitations with the same antibody. Precipitation with a crossreacting human beta 2-microglobulin (beta 2-m) specific antibody confirmed a class-I-like structure associated with beta 2-m on M' positive red cells and the absence of such a structure on M' negative red cells. Sequential precipitations gave analogous results. Proteolytic degradation by papain and V8 protease did not reveal any substantial difference between red and white M'-A16 positive cells, but a slight difference in the pI of the immunoprecipitable components of red and white cells was observed. All together, this indicates that either the blood group antigen M' is the BoLA-A16 class I antigen or M' and BoLA-A16 are two different class I polypeptides with the same relative mass, sharing identical epitopes and both associated with beta 2-m. Comparable results were obtained with M1 and BoLA-A24. PMID- 7486248 TI - Random amplified polymorphic DNA in cattle and sheep: application for detecting genetic variation. AB - The present study investigated the use of the random amplified polymorphic DNA (RAPD) method to detect genetic variation in cattle and sheep. The animals studied consisted of samples from five Finnish cattle breeds: native Eastern (18 animals), Northern (24), Western Finncattle (24), Finnish Ayrshire (24), and Finnish Friesian (18); as well as a white (6 animals) and a grey (9) colour type of Finnsheep. The cattle and sheep populations were analysed with 11 and 13 RAPD primers demonstrating the most repeatable amplification pattern. Two out of ten RAPD fragments tested by cross hybridization showed homology between the two species. The RAPD method did not prove efficient for finding new polymorphisms in either species, because we found only three polymorphic RAPD markers for cattle and seven markers for sheep with different allele frequencies between the breeds. Although there is a greater presence of polymorphic RAPD markers in sheep, according to the similarity indices the sheep populations showed a higher degree of homogeneity than the cattle breeds. However, the interbreed and intrabreed similarity indices for cattle did not suggest any significant differentiation of the Finnish breeds, contrary to earlier results based on blood group and protein polymorphism. PMID- 7486249 TI - SSCP analysis of pig mitochondrial DNA D-loop region polymorphism. AB - The sequence polymorphism that occurs in the mitochondrial DNA (mtDNA) displacement (D)-loop region is useful as a cytoplasmic DNA marker. We cloned the mtDNA D-loop regions of five breeds of pig by polymerase chain reaction (PCR) and determined their sequences. The sequence diversities in D-loop regions among five breeds of pig were located in the starting area of heavy-strand replication. From these sequences, we designed primers for PCR-mediated single-strand conformation polymorphism (PCR-SSCP) analysis that amplified the most polymorphic 227 bp fragment of the D-loop region. The results of PCR-SSCP analysis clearly showed that four types of polymorphism (A to D) are found in Landrace (A), Large White (A, B), Duroc (A), Gottingen miniature pig (B) and Meishan (C, D). The same polymorphisms were also detected from each porcine embryo by this method. Our results show that PCR-SSCP analysis is useful in detecting polymorphisms in the D loop region of pigs and pig embryos. PMID- 7486250 TI - Polymorphic CAC/T repetitive sequences in the pig genome 1. AB - Three genomic clones were isolated from a size-selected pig DNA library by hybridization with a DNA-fingerprint probe. Analysis at the sequence level revealed that all three clones contain interrupted stretches of triplet repeats mainly composed of CAC and CAT triplets. Evaluation of the corresponding loci for polymorphism by Southern blot hybridization showed considerable length variation. For two loci the polymorphism was also demonstrated by polymerase chain reaction (PCR) amplification. The PiGMaP reference pedigree was typed for all three loci. PMID- 7486252 TI - Genotyping of Bison bonasus kappa-casein gene following DNA sequence amplification. AB - DNA isolated from 30 Bison bonasus blood samples was used for amplification of a kappa-casein gene fragment to detect possible polymorphism. Bovine kappa-caseins exist in A, B and E variants. In the European bison from Poland, we have found only BB genotypes. Sequencing of the polymerase chain reaction product revealed further polymorphisms typical for B. bonasus. PMID- 7486251 TI - Sheep gene mapping: assignment of ALDOB, CYP19, WT and SOX2 by somatic cell hybrid analysis. AB - Twenty-four hamster-sheep hybrid cell lines representing eleven ovine synteny groups were used to make syntenic assignments for seven loci ALDOB (aldolase B, fructose biophosphate); AMH (anti-Mullerian hormone); CYP19 [cytochrome P450 aromatase, subfamily XIX (aromatization of androgens)]; WT (Wilms' tumour gene); SOX2 (SRY-related HMG-box gene 2); FSHB (follicle-stimulating hormone, beta polypeptide); and SRY (sex region of Y chromosome). These loci were assigned to synteny groups U11(chr2) (ALDOB); U19 (AMH); U3(chr7) (CYP19); and to chromosome 15 (WT) and 1 (SOX2). SRY defines the hybrids containing the Y chromosome. PMID- 7486253 TI - Mapping the soluble angiotensin binding protein (ABP1) locus to porcine chromosome 16. AB - A highly polymorphic (AT)nTm microsatellite located in a PRE1 SINE element in the 3'UTR of the soluble angiotensin binding protein (ABP1) gene has enabled the linkage mapping on the PiGMaP reference families of the ABP1 gene to porcine chromosome 16, to a region homologous with the short arm of human chromosome 5. PMID- 7486254 TI - Assignment of the growth hormone receptor gene to bovine chromosome 20 using linkage analysis and somatic cell mapping. AB - A polymorphism was identified in the bovine growth hormone receptor (GHR) gene by digesting polymerase chain reaction (PCR) products with the restriction enzyme Alul. Two alleles were segregating in cattle of Bos indicus descent, but one allele appears to be fixed in Bos taurus cattle. GHR was localized to bovine chromosome 20 using bovine-rodent hybrid cell lines and linkage analysis. PMID- 7486255 TI - Expression of multiple DQB genes in Bos indicus cattle. AB - Polymorphism of expressed bovine major histocompatibility complex (MHC) class II DQB genes was investigated in a group of nine MHC-homozygous Kenya Boran cattle (Bos indicus). DQB second exon fragments were amplified by reverse transcription polymerase chain reaction of total mononuclear cell RNA, cloned and sequenced. While a single DQB sequence was obtained from some animals, two DQB exon 2 sequences were found in others, implying expression of duplicated DQB genes. Two pairs of duplicated DQB genes were found in this group of homozygotes, and sequence analysis showed that both pairs contained distinct DQB alleles. One DQB duplication was observed in two related animals with the same MHC haplotype (F100 and F188), while the second was detected in three animals (F187, G166 and G277) that expressed the same class II DRB3 and DQB alleles but had different class I (BoLA-A) types, suggesting that this DR/DQ haplotype may be widespread in Boran cattle. PMID- 7486256 TI - A monoclonal antibody to swine erythrocytes recognizes the B blood group on the major glycophorin. AB - Recently monoclonal antibodies (mAbs) to swine red blood cells have been described. One of them (1AC11) was specific for the major swine glycoprotein with a molecular weight of 45 kDa and another mAb, 2G2, recognized the Ba allele in the B system of swine blood groups. Immunoblotting experiments to characterize the mAb 2G2 indicated that it reacts with an antigen of 45 kDa, present on the aqueous phase, glycophorin fraction, of swine red blood cells with the Ba allele and does not react with BbBb homozygous cells. The antigen recognized by 2G2 has the same characteristics as the major glycophorin recognized by 1AC11, so we can conclude that the B system of the swine blood group is on the major glycophorin of swine erythrocyte membranes. PMID- 7486257 TI - A set of polymorphic DNA microsatellites useful in swamp and river buffalo (Bubalus bubalis). AB - DNA microsatellites have found widespread application in gene mapping, pedigree determination and population genetics. In closely related species such as bovids, heterologous polymerase chain reaction (PCR) primers may in some cases be used, bypassing the need to isolate and characterize microsatellite-containing sequences and design PCR primers. We report on the ability of a set of eighty bovine derived DNA microsatellite primers to amplify sequences in the two types (swamp and river) of water buffalo (Bubalus bubalis). Number of alleles and per cent heterozygosities in a large number of animals were determined on a subset of microsatellite loci selected on the robustness of the primers. These loci will form the basis of a set of polymorphic DNA markers for use in water buffalo. PMID- 7486259 TI - Five polymorphic bovine microsatellite loci: IDVGA-62A, IDVGA-71, IDVGA-82, IDVGA 88, IDVGA-90. PMID- 7486258 TI - Alignment of the PiGMaP and USDA linkage maps of porcine chromosomes 2 and 5. AB - The PiGMaP and USDA porcine linkage maps for chromosomes 2 and 5 have been aligned by typing five USDA microsatellite markers from chromosomes 2 and 4 from chromosome 5 on the PiGMaP reference families. The markers in the two maps can be successfully aligned except for Sw395 on chromosome 2, which is the end-most marker in the USDA map 22 cM remote from the next marker, but which maps to a more central location and in the same position as Sw776 in the PiGMaP families. The mapping of four additional chromosome 5 markers has enabled amalgamation of the two previously separate PiGMaP linkage groups assigned to chromosome 5 and has more than doubled the length of its map. The USDA map of chromosome 5 is considerably shorter than the revised PiGMaP version, particularly between DAGK and Sw1071, where the corresponding lengths are 9 cM versus 33 cM. PMID- 7486260 TI - Dinucleotide repeat polymorphism at the porcine TNFA locus. PMID- 7486261 TI - An EcoRI polymorphism at the amylase 1 (AMY1) locus in cattle. PMID- 7486262 TI - A polymorphic dinucleotide repeat microsatellite in Atlantic salmon, Salmo salar (SSOSL436). PMID- 7486263 TI - A PCR-based diagnostic test for the endogenous retroviral element ev4 of chickens. PMID- 7486264 TI - Polymorphic sheep microsatellites at the McM2, McM131, McM135, McM136, McM140, McM200, McM214, McM373, McM505, McM507 and McM512 loci. PMID- 7486265 TI - Ovine dinucleotide repeat polymorphism at the McM15, McM152, McM159, McM164 and McM210 loci. PMID- 7486267 TI - UCD001: a bovine cosmid containing a polymorphic dinucleotide repeat maps to chromosome 4. PMID- 7486266 TI - Ovine dinucleotide repeat polymorphism at the CSRD226 and CSRD232 loci. PMID- 7486268 TI - A Bgl/II RFLP at the porcine calpastatin (CAST) locus. PMID- 7486269 TI - A PstI RFLP at the porcine beta-myosin heavy chain (MYHCB) locus. PMID- 7486270 TI - [Neuropathy in intensive care: a challenge for the future]. PMID- 7486271 TI - [Comparison of vascular effects of magnesium sulfate and nicardipine during extracorporeal circulation]. AB - OBJECTIVE: To evaluate the hypothesis that magnesium sulphate (SO4Mg), usually administered for protecting the myocardium and decreasing the rate of arrhythmias in cardiac surgery, was able to control the hypertensive peaks occurring during cardiopulmonary bypass (CPB), as efficiently as nicardipine (N). STUDY DESIGN: Randomized controlled trial. PATIENTS: Forty patients were allocated into two groups when hypertensive peaks occurred during CPB. The patients of the N group were then given nicardipine 0.016 mg.kg-1 and those of the SO4Mg group received magnesium sulphate 50 mg.kg-1. METHODS: Anaesthesia technique was identical and during normothermic CPB the flow remained constant at 2.4 L.min-1, during the 10 min following N or SO4Mg administration. The usual haemodynamic variables were monitored. RESULTS: Both N and SO4Mg decreased significantly the MAP and the SVR over the 9 min following their administration. The decrease was more marked after SO4Mg. As the flow of the pump was unchanged after their injection the decrease can be attributed to the relaxing effect of these agents on the smooth vascular muscles. CONCLUSIONS: SO4Mg permits to treat hypertensive peaks occurring during CPB as efficiently as N. Three grammes of SO4Mg have an hypotensive effect equivalent to that of 1 mg of nicardipine. However repetitive injections of SO4Mg cannot be recommended because of the risk for hypermagnesemia. PMID- 7486272 TI - [Comparison of effects of aprotinin and tranexamic acid on blood loss in heart surgery]. AB - OBJECTIVES: To compare the efficacy of aprotinin (APR) and tranexamic acid (TRA) in reducing blood loss and transfusion requirements after cardiac surgery under extracorporeal circulation (ECC). STUDY DESIGN: Randomized controlled trial. PATIENTS: One hundred and four adults undergoing either coronary artery bypass grafting (CABG) (n = 55), or aortic valve replacement (AVR) (n = 49), allocated into three groups. METHODS: a) APR group (23 CABG and 20 AVR) received aprotinin, 2 x 10(6) KIU (280 mg) after induction, followed by an infusion of 0.5 x 106 KIU.h-1 (70 mg.h-1) until chest closure, with a supplement to the oxygenator prime of 2 x 10(6) KIU; b) TRA group (22 CABG and 19 AVR) received tranexamic acid, 15 mg.kg-1 between the injection of heparin (400 IU.kg-1) and the beginning of ECC, 15 mg.kg-1 after protamin injection (1.3 mg/100 IU of heparin); c) CTR group (10 CABG and 10 AVR), the control group, was not treated with an antifibrinolytic agent. The amount of blood collected from the chest tube drainage was measured at admission to ICU, as well as 4, 8 and 18 h after the insertion of drains and at the time of their removal. Packed red cells where given when the haematocrit was under 20% during ECC, 25% at the end of surgery and 30% after extubation. RESULTS: The blood loss was lower in APR group (834 +/- 448 mL) than in TRA group (1015 +/- 409 mL) (P = 0.009), and in CTR group (1416 +/- 559 ML) (P = 0.004). The rates of transfused patients in groups APR, ATR and CTR were 35, 37 and 60% respectively and the numbers of units administered per patient were 0.8, 0.8 and 1.7 respectively. In AVR cases, APR and TRA had a similar efficacy. In CABG cases, only aprotinin decreased postoperative bleeding. However there was no difference between APR and TRA concerning the transfusion requirements. In CABG cases the ECC was of shorter duration and blood loss was 1127 +/- 540 mL vs 894 +/- 422 mL in AVR cases (P = 0.03). CONCLUSIONS: Both APR and TRA decrease blood loss. APR is more efficient after CABG than TRA as far as blood loss is concerned, whereas the transfusion requirements are similar. As APR is about 100 times more expensive and carries a risk for allergic reactions, its use in a high dose regimen is only recommended for reoperations, in patients treated with salicylates and in case of sepsis. PMID- 7486273 TI - [Pharmacokinetics of lidocaine and bupivacaine after peribulbar block with additional hyaluronidase]. AB - OBJECTIVE: To assess the time course of plasma concentrations of lidocaine and bupivacaine associated with hyaluronidase for peribulbar block. STUDY DESIGN: Prospective study. PATIENTS: Ten patients (mean age = 71 +/- 11 yrs, mean weight 63 +/- 10 kg) scheduled for cataract surgery with lens implantation. METHOD: Lidocaine 2% (5.5 mL = 110 mg) and bupivacaine 0.5% (5.5 mL = 27.5 mg) associated with hyaluronidase (80 IU) were injected supra and infra-orbitally, in patients premedicated with midazolam. Blood samples wer0 collected at constant time intervals from the end of infiltration until the 6th hour. The plasma concentrations of local anesthetics were measured with the HPLC technique. RESULTS: The median plasma peak concentration was 1.74 mg.L-1 after 10 min for lidocaine, and 0.52 mg.L-1 after 7.5 min for bupivacaine respectively. CONCLUSIONS: The similar delays of occurrence of peak concentrations confirm that liposolubility is not the only factor of diffusion of local anaesthetics from the periocular fat into the blood stream. The peak concentrations are far below the alleged toxic concentrations. When associated with hyaluronidase, the peak concentrations occur as rapidly as after endotracheal or paracervical administration. PMID- 7486274 TI - [Cerebrovascular reactivity to CO2 during general anesthesia maintained with either isoflurane-N2O or propofol-N2O. A comparative study by transcranial Doppler velocimetry]. AB - OBJECTIVES: To compare, using transcranial doppler velocimetry (TDV), the cerebral blood flow velocity and CO2 reactivity during general anaesthesia maintained with either isoflurane-N2O-O2(IF) or propofol-N2O-O2 (PF) in adults with a normal brain. STUDY DESIGN: Nonrandomized controlled trial. PATIENTS: Forty ASA I patients (mean age 41 +/- 13 yrs, 15 F/35 M) undergoing surgery of the lumbar spine in prone position. The first 20 were allocated into the IF group and the subsequent 20 into the PF group. METHODS: General anaesthesia was induced with midazolam (0.05 mg.kg-1), phenoperidine (0.03 mg.kg-1), thiopentone (5 mg.kg 1), vecuronium (0.1 mg.kg-1) and maintained with N2O (50 vol %) and O2 (50 vol %) and either isoflurane (0.8 < Fet < 1 vol %) in IF group or propofol (6 mg.kg-1.h 1) in PF group. The vascular reactivity was assessed with velocimetry measurements (Angiodine DMS, with a probe transmitting a 2-MHz pulsed wave) of flow in the middle cerebral artery at a given PetCO2 (obtained by adjustments of VT) during systole (SV) and diastole (DV). Three measurements were made: at T1 (PetCO2 = 30 +/- 2 mmHg), at T2 (PetCO2 = 40 +/- 2) and at T3 (PetCO2 = 30 +/- 2 mmHg). RESULTS: In the IF group, VS increased by +32% at T2 (P = 0.006) with an increase of + 3.4 %/1 mmHg of PetCO2. Similarly, in the PF group VS increased by + 31 % at T2 (P < 0.0001) with an increase of 2.9 %/1 mmHg of PetCO2. In both groups the VS returned to baseline values at T3. In the IF group, VD increased by + 66% at T2 (P < 0.0001), with an increase of + 7%/1 mmHg of PetCO2. Similarly in the PF group, VD increased by + 61% (P < 0.0001) with an increase of + 5.7%/1 mmHg of PetCO2. In both groups the VD returned to baseline values at T3. CONCLUSIONS: During anaesthesia maintained with either isoflurane-N2O-O2 or propofol-N2O-O2, a change in PetCO2 results in similar changes in VS and VD. These anaesthetic agents preserve the cerebrovascular reactivity of the normal brain. The results of this study are in accordance with those obtained with other reference techniques in healthy volunteers. Transcranial doppler velocimetry can be a useful noninvasive tool of clinical research in neuroanaesthesia. PMID- 7486275 TI - [Retrograde percutaneous catheterization of the internal jugular vein in head injured patients]. AB - The monitoring of jugular venous oxygen saturation and lactate concentration in order to detect cerebral ischaemia or hyperaemia requires the insertion of a fibreoptic catheter into the upper bulb of the internal jugular vein. With dissection studies we have defined superficial anatomical landmarks which are constant and easily palpable, namely the mastoid process and the sternocleidomastoid muscle. This technique does not require a rotation of the head. The puncture site is located at the top of a triangular area between its sternal and clavicular insertions. At this site the jugular vein is rather superficial. This study reports our experience of the retrograde catheterisation of the jugular vein in twelve severely head injured patients. PMID- 7486276 TI - [Myocardial ischemia and anesthesia]. AB - Patients with coronary artery disease are particularly at risk perioperatively, as myocardial infarction, unstable angina, severe arrhythmia and cardiac death may occur. These events are often preceded by prolonged silent myocardial ischaemia (MI). Moreover, perioperative MI predicts long-term adverse cardiac outcome. Therefore, it is logical to prevent and treat MI. However, the detection of perioperative MI is difficult because of low sensitivity of ST-segment monitoring, low specificity of echocardiography and insufficient availability of equipment for its monitoring. A pragmatic approach is described, including preoperative consideration of myocardial revascularization prior to non-cardiac surgery and perioperative administration of antianginal agents; the effects of clonidine are discussed as well. The role of anaesthetic techniques and normovolaemic haemodilution is considered. MI episodes may be prevented or their duration may be shortened by treating tachycardia, hypotension and, possibly, hypertension. The risks of MI are particularly high during the postoperative period because increased global oxygen consumption associated with recovery, ventilator weaning, shivering and pain may lead to tachycardia and increased ventricular load. These factors must be taken into account in order to prevent MI and improve postoperative cardiac outcome. PMID- 7486277 TI - [Invasive aspergillosis in intensive care]. AB - Invasive pulmonary aspergilloses occur in patients with antineoplasic chemotherapy, mainly when associated with a prolonged neutropenia, in transplanted patients with continuous corticotherapy and less frequently in immunocompetent surgical patients. The clinical features are those of an acute infective pneumonia, not responding to antibiotherapy. Radiologic signs are often non specific. Diagnosis is obtained with bronchoalveolar lavage in which Aspergillus is found both at direct examination and in culture. Serological tests are of little interest for the diagnosis of invasive aspergillosis. Extrapulmonary locations such as sinusitis, cutaneous or brain abscesses occur in 20% of cases. The gold standard of treatment is intravenous amphotericin B which elicits an acute reaction often followed by a nephrotoxic effect which can be decreased by fluid loading with saline. Oral itraconazole administration can follow the initial treatment with amphotericin B. The mortality rate remains high and an early diagnosis and an appropriate treatment are essential. PMID- 7486278 TI - [A cause of difficult weaning from artificial respiration: polyneuropathy of intensive care]. AB - Three cases of Critically Illness Polyneuropathy (CIP) are reported. The difficulty of weaning the patients from the ventilator, whereas sepsis and MOF had been successfully treated, was the main feature in the three cases. The diagnosis was established by eletromyogram, showing a primary axonal denervation of peripheral nerve fibres, which was confirmed by a pathological study in one patient. PMID- 7486279 TI - [Secondary neuromuscular deficiencies in critical care patients]. AB - The authors report three cases of neuromuscular deficits occurring after a variable time of mechanical ventilation in ICU patients suffering from acute respiratory failure requiring sedation and neuromuscular blockade by pancuronium bromide. The clinical features of these deficits were similar : flaccid quadriparesia slowly reversible without sensory loss and difficult weaning from ventilation for two patients. The paraclinical investigations (measurements of creatine kinase, electrophysiological examination) permitted to distinguish two diseases: critical illness polyneuropathy (in patient with ARDS) and acute myopathy (in other patients with severe asthma) caused by corticosteroids and potentiated by neuromuscular blocking agents. This myopathy differs from the chronic myopathy caused by long term corticotherapy. After a status asthmaticus treated with corticosteroids and mechanical ventilation under muscle relaxants, creatine kinases should be measured in plasma and a neurological examination be performed. The value of neuromuscular blockade monitoring for the prevention of this myopathy should be assessed. PMID- 7486280 TI - [Post-traumatic dissection of the aortic isthmus with favorable outcome without surgical treatment. Value of transesophageal echocardiography]. AB - We report a case of intimal rupture of aortic isthmus in a severe polytrauma patient diagnosed with transoesophageal echocardiography (TEE). TEE findings were as follows: localized intimal tear without increased aortic diameter or pseudoaneurysm, absence of mediastinal haematoma or false lumen. CT scan and MRI confirmed the diagnosis. Because of the associated injuries, surgery was delayed. Conservative management was successful and the patient was discharged without aortic surgery. A follow up TEE, 3 and 6 months later showed a thrombosis of the intimal tear without false aneurysm. This case underlines the value of TEE in patients with thoracic trauma, for surgical or nonsurgical management of aortic rupture and for the follow up of the patients. PMID- 7486281 TI - [Bilateral bronchial rupture: problems of respiratory management]. AB - The authors report the case of bilateral bronchial rupture in a 39-year-old multiple trauma patient. During the thoracotomy for right main bronchus repair, a partial left bronchial rupture was recognized because of severe hypoxaemia after left selective intubation. PMID- 7486282 TI - [Tracheal compression by mediastinal mass in children. Value of fiberoptic tracheobronchoscopy for anesthetic management]. AB - Due to the action of the drugs and the position of the patient, general anaesthesia in a child with a mediastinal tumor can complete the obstruction of the trachea and/or the main bronchi and possibly result in irreversible respiratory failure. The risk is directly linked to the degree of lumen amputation of the trachea, usually evaluated by CT-scan: there is a risk of accident if the degree of lumen amputation is greater than 30%. The case reports underline the value of the flexible fibreoptic bronchoscopy under local anaesthesia. This examination, as well as the CT-scan, allows to evaluate the degree of tracheal compression. Moreover, it carries a lower risk than the CT scan which requires, in a small child, either general anaesthesia or heavy premedication which could lead to asphyxia. Total obstruction most often occurs with anterior mediastinal tumors. However, even posterior or lateral tumors can extend to an anterior position, especially in the young child. If the obstruction of the lumen is greater than 30% and if general anaesthesia is required, the fibreoptic bronchoscopy allows endotracheal intubation under local anaesthesia before general anaesthesia. We report 5 cases which illustrate the risk of tracheal compression during general anaesthesia in patients with a mediastinal tumor and substantiate the advantages of a flexible fibreoptic bronchoscope in the management of these patients before and during general anaesthesia. PMID- 7486283 TI - [Delayed low back pain after spinal anesthesia]. AB - A 50-year-old man had an inguinal hernia repair under spinal anaesthesia with bupivacaine. On the 2nd postoperative day, he complained of backache and paresis at the posterior part of the lower extremities, well relieved by non-steroidal anti-inflammatory drugs. On the 6th postoperative day, he came back to hospital, because of low back pain associated with a heavy feet sensation. The hypothesis of a neurological complication of the spinal anaesthesia was considered. The interview of the patient revealed a history of lumbar disk disease, not reported during the preoperative visit. After an in depth clinical examination, two causes seemed possible: subarachnoid haematoma and lumbar disk protusion. Against the first diagnosis were the initial clinical signs. However, in many cases, objective neurological deficit arise too late to allow efficient neurosurgical treatment. Thus, a MRI examination was performed which is non invasive in comparison with a computed tomography myelogram. In our patient, it did not detect a true lumbar disk protusion, but a simple degenerative disease of the L5 S1 disk. In suppressing the lumbar lordosis, spinal anaesthesia probably allowed a distension of spinal capsules and tendons, responsible for the troubles. PMID- 7486284 TI - [Cephalic cancer pain controlled by intraventricular administration of morphine and clonidine]. AB - A 47-year-old woman suffered from a pharyngeal cancer causing pain only poorly relieved by high doses of oral morphine. Oral administration was switched to the intracerebroventricular (ICV) route but pain relief was only transient despite a daily dosage up to 1.5 mg of morphine. Finally an effective pain relief was achieved with increasing doses of clonidine (5 to 30 micrograms) in combination with morphine (1.5 mg) by ICV route. Neither arterial hypotension nor sedation occurred and the patient recovered transiently a better quality of life. PMID- 7486285 TI - [Headache after spinal anesthesia treated with sumatriptan]. PMID- 7486286 TI - [Toxicity of gentamycin administered by autotransfusion of lost blood after prosthetic knee surgery using gentamycin cement]. PMID- 7486287 TI - [Risk of recurarisation after postoperative autotransfusion]. PMID- 7486288 TI - [Peroperative herniation of a laryngeal mask cuff]. PMID- 7486290 TI - [60th anniversary of the 1st French Society of Anesthesia]. PMID- 7486291 TI - [13th Consensus Conference on Intensive Care and Emergency Medicine. Assisted respiration during acute decompensation in chronic respiratory failure in adults (excluding neuromuscular diseases and weaning). 2 December 1994, Strasbourg]. PMID- 7486292 TI - [The impact factor: "audimat", a valuable quality indicator for scientific journals?]. PMID- 7486289 TI - [Information wanted: Who knows Hood? A hint of a track!]. PMID- 7486293 TI - [Comparison of extrapleural and intrapleural analgesia with bupivacaine after thoracotomy]. AB - OBJECTIVE: To compare the analgesic and the ventilatory effects as well as blood concentrations of bupivacaine, administered either in the extrapleural or interpleural space after posterolateral thoracotomy. STUDY DESIGN: Randomized clinical trial. PATIENTS: Twenty ASA class I and II patients, scheduled for elective thoracic surgery were randomly allocated either in the IP group (catheter inserted into the interpleural space) or the EP group (catheter inserted in extrapleural position, paravertebrally above the posterior parietal pleura). METHODS: In the catheter, inserted at the Th4 level at the end of the surgical procedure, 20 mL of 0.5% bupivacaine were injected after full recovery from anaesthesia, with the thoracic drains clamped for 30 min. The injection was repeated every six hours. Pain was evaluated after a deep inspiration with a visual analog scale (VAS), before and 1.3 and 6 hours after the injection. Analgesia was considered as effective if the VAS score at the end of the first hour was less than 30 mm. Otherwise 0.1 mg.kg-1 of morphine was administered subcutaneously. The forced vital capacity (FVC) and the forced expiratory volume one second (FEV1) were measured preoperatively and on 1st (D1) and 2nd postoperative Day (D2). Blood samples for measurements of plasma bupivacaine concentrations were obtained at 5, 10, 20, 30, 60, 90, 120, 150, 180 and 250 min respectively after the first injection. RESULTS: Bupivacaine provided a more rapid, deep and prolonged analgesia by extrapleural than by interpleural route. Analgesia was effective in 9 patients in EP group vs 4 patients in IP group (P < 0.05). Morphine requirements were 4 +/- 8 mg in EP group vs 17 +/- 10 mg in the IP group (P < 0.05). The FVC and FEV1 values were similarly decreased on D1, but recovery was better in EP group on D2 (P < 0.05). Bupivacaine peak concentrations in plasma were lower in EP group (0.86 +/- 0.42 microgram.mL-1) than in IP group (1.63 +/- 1.44 micrograms.mL-1), however the difference was not significant. CONCLUSIONS: Extrapleural administration of bupivacaine provides better analgesia as the anaesthetic agent comes in closer contact with intercostal nerves and with lower risk of loss of agent through the pleural drainage. Therefore its use is recommended preferentially over the interpleural route for analgesia after posterolateral thoracotomy. PMID- 7486294 TI - [Cervicofacial cellulitis of dental origin and tracheal intubation]. AB - OBJECTIVES: To evaluate the difficulty of intubation in relation with the localisation and spread of cervico-facial cellulitis of odontogenic origin and to recognize the optimal technique of intubation in such circumstances. STUDY DESIGN: Prospective clinical open study. PATIENTS: Hundred patients, including 16 children, undergoing surgical drainage of a cervico-facial cellulitis of odontogenic origin under general anesthesia were studied. METHODS: Difficulty of intubation was evaluated with the following four criteria: active mouth opening in the awake patient, Mallampati's classifying system, presence of trismus, clinical and radiological control of localisation and extension of the cellulitis (mandibular, maxillar or mouth floor). In case of a foreseen difficult intubation, a fibrescope was used in the awake patient. Otherwise the endotracheal tube was inserted after administration of propofol (3 mg.k-1) and alfentanil (10 to 20 micrograms.kg-1). A Cormack's grading was performed during intubation. RESULTS: Mouth opening depended on the localisation of the cellulitis. Trismus occurred more often with mandibular than maxillary localisations. Trismus and a Mallampati's class > 2 were associated with difficulty in intubation (Cormack's grade > 2), except in maxillary localisations. CONCLUSIONS: The localisation of cellulitis of odontogenic origin is responsible for the difficulty grade of intubation. Awake fibreoptic intubation should be systematically performed in patients with a floor of the mouth cellulitis to reduce the risk of rupture of the abscess by a laryngoscope blade. As trismus associated with mandibular localisations is not relieved by general anaesthesia, awake fibreoptic endotracheal intubation should be preferred. PMID- 7486295 TI - [Endotracheal intubation under propofol with or without vecuronium]. AB - OBJECTIVE: In order to test the hypothesis that under the association propofol alfentanil-IV lidocaine the trachea could be intubated easily without an additional muscle relaxant, this study compared the intubation conditions when this association was combined or not with vecuronium. STUDY DESIGN: Randomized comparative trial. PATIENTS: The study included 152 young adults classified as ASA physical class I and Mallampati presentation grade 1, randomly allocated either into Vecu+ group or Vecu0 group, depending on whether vecuronium was co administered or not. METHODS: All patients received midazolam 0.05 mg.kg-1 i.v., one minute before induction. Those of group Vecu0 were given successively within two minutes: alfentanil 0.03 mg.kg-1, lidocaine 1.5 mg.kg-1 i.v. and propofol 2.5 mg.kg-1. Patients of group Vecu+ received similar doses of alfentanil and propofol as well as vecuronium 0.08 mg.kg-1. The endotracheal tube was inserted one minute after induction in the patients of Vecu0 group, and after three minutes in those of the Vecu+ group. During intubation, scores of mouth opening, glottis opening and coughing were established, in order to assess intubation conditions. RESULTS: Similar convenient intubating conditions were obtained in both groups (in 97% of patients in Vecu+ group vs 95% of those in Vecu0 group). In the latter, the glottis opening was less pronounced. CONCLUSIONS: In young healthy adults, without anaesthetic risk (emergency, full stomach) and without foreseen difficult intubation, the endotracheal tube can be inserted in convenient conditions without a muscle relaxant, under the association propofol alfentanil-lidocaine iv. PMID- 7486296 TI - [Effects of platelet-rich plasma on hemostasis and transfusion requirement in vascular surgery]. AB - OBJECTIVE: To assess the effect of intraoperative autologous platelet-rich plasma (PRP) transfusion on haemostasis, blood loss and blood requirements during vascular surgery. STUDY DESIGN: Randomized clinical trial. PATIENTS: Twenty patients undergoing elective abdominal infrarenal aortic aneurysmectomy, using autologous transfusion techniques (predonation programme and/or preoperative normovolaemic haemodilution and/or intraoperative use of a cell-saver), were randomly allocated either into the PRP group (n = 10) or the Control group (n = 10). METHOD: In patients of PRP group, 10 mL.kg-1 of PRP were obtained over 40 to 50 min, prior to induction of anaesthesia, and compensated simultaneously with an equivalent amount of hydroxyethyl starch. Each PRP unit was transfused to its donor after aortic declamping. Blood samples were obtained before induction, before incision, at wound closing and at the end of PRP unit transfusion for determination of biological variables. RESULTS: The PRP units transfused in the patients of PRP group contained 755 +/- 117 mL of plasma with a platelet count of 62 +/- 31 G.L-1. The intra and postoperative blood losses were similar in both groups (1622 +/- 758 and 233 +/- 322 mL respectively in PRP group vs 1890 +/- 1331 and 291 +/- 303 mL respectively in Control group). In both groups, three patients required an additional transfusion of homologous blood. The results of biological tests (haematocrit, platelet and white cell counts, prothrombin time, aPTT, thrombin time, fibrinogen, D-dimers, proteins, calcium) were also similar between groups at the various times of sampling. The reinfusion of the PRP unit did not increase the platelet count. CONCLUSIONS: This study demonstrates that intraoperative infusion of autologous PRP does not decrease blood loss and homologous transfusion requirements in patients undergoing elective abdominal infrarenal aortic aneurysmectomy. This result can be related to the relatively moderate enrichment in platelets obtained with the centrifugation speed used in this study. PMID- 7486297 TI - [Effects of midazolam on respiratory drive in healthy volunteers]. AB - OBJECTIVE: To compare the effects of a sedative dose of midazolam on mean inspiratory flow (VT/TI = index of central respiratory activity), known as being decreased by midazolam and the intercostal muscle activity, known as being increased by this agent. STUDY DESIGN: Laboratory study. PATIENTS: Seven healthy volunteers. METHODS: After assessment of baseline values of ventilatory variables and intercostal electromyographic activity (in arbitrary units), midazolam 0.1 mg.kg-1 was administered by iv route. The measurements were repeated after 5 and 10 min, and finally 2 min after the i.v. injection of flumazenil 1 mg. RESULTS: Midazolam decreased VE and VT. Similarly VT/TI ratio decreased from 0.44 +/- 0.04 (baseline value) to 0.26 +/- 0.03 (5 min) and 0.3 +/- 0.03 L.s-1 (10 min later) respectively (P < 0.05). Conversely, midazolam increased the intercostal electromyographic activity from 4.0 +/- 0.7 (baseline value) to 26.5 +/- 16.6 (5 min) and 28.4 +/- 16.6 U (10 min later) respectively (P < 0.05). Within 2 min after flumazenil administration all variables returned to baseline values. CONCLUSIONS: The decrease of VT/TI ratio is probably linked to increased resistances in the upper airways. This ratio cannot act as an indicator of respiratory drive during sedation or anaesthesia. The assessment of the ventilatory effects of benzodiazepines must be based simultaneously of the various other indicators of the ventilatory drive, as these agents act on the different stages of the ventilatory cycle and therefore cannot be characterized by a unique variable. PMID- 7486298 TI - [High frequency jet ventilation combined with conventional mechanical ventilation in the treatment of adult respiratory distress syndrome]. AB - Better understanding of the physiopathology of ventilatory mechanisms associated with ARDS and the recent re-evaluation of the iatrogenic potential of mechanical ventilation (MV) brings us closer to the best suited ventilatory mode for these patients. In severely ill ARDS patients, only a small lung volume is ventilated, and remains available for the totality of the gas exchanges (baby lung concept). The goal of MV is to restore and maintain an optimal exchange volume while limiting mechanical agression of the lung tissue. Analysis of the ARDS related pressure-volume relationship (P/V) is helpful in specifying the tolerable limits of the ventilatory pressure regimen. The lower limit (end expiratory pressure) must be kept above the lower inflexion point of the curve, in order to increase the ventilated lung volume and avoid distal airway collapse. Under this limit, gas exchanges are altered by the shunt effect, and shear stress lesions result from the repeated opening and closing of the distal airways. The upper limit (end inspiratory pressure) must be situated below the upper inflexion point of the curve, in order to avoid lesions resulting from surdistension of the alveolocapillary membranes and barotraumatisms. The only way to position MV in such a narrow pressure window, is to greatly reduce the tidal volume (VT). Though CO2 retention would inevitably occur under conventional MV conditions, high frequency ventilation (HFV) seems better adapted to these theoreotical objectives; small VT's are injected under a limited amplitude pressure regimen and a satisfactory CO2 clearance is maintained. This ventilatory mode, existing since more than 15 years, has recently benefited from many technical improvements as well as the concept of oscillating the ventilation around a selected mean pressure in the central region of the P/V curve. In the past, HFV was applied using low pressure regimens, situated below the lower inflexion point of the curve. The resulting failures are, a posteriori, explained by insufficient lung volumes, unable to maintain adequate gas exchanges. Current work is aimed at re evaluating HFV, using higher mean airway pressure levels. Combined HFV is another advance towards the theoretical goal of restoring and maintaining optimally ventilated lung volumes. Though HFV alone can maintain lung volumes oscillating around a mean value, it cannot re-expand atelectatic areas. The small VT's used are insufficient to overcome these area's elevated opening pressures. Volume recruitment by periodic hyperinflations, or sighs, though generally considered useless in conventional MV, have been shown to be of great benefit in HFV.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 7486299 TI - [General anesthesia in a patient with catecholaminergic polymorph ventricular tachycardia]. AB - A patient suffering from a catecholaminergic polymorph ventricular tachycardia treated with nadolol was scheduled for dental extractions under general anaesthesia. During induction, whereas the basal heart rate was at 30-35 b.min-1, occurred an episode of bidirectional ventricular tachycardia probably induced by the administration of pancuronium. PMID- 7486300 TI - [False aneurysm of the brachiocephalic artery following subclavian vein catheterization]. AB - The difficult insertion of a right subclavian catheter in a 63-year-old patient resulted in a pseudoaneurysm of the brachiocephalic artery requiring a resection suture. The preventive measures of that complication are reminded. PMID- 7486301 TI - [Brachiocephalic trunk erosion by a tracheotomy cannula]. AB - We report the case of a 27-year-old severe head trauma patient who developed an erosion of the brachiocephalic artery, 19 days after the insertion of a tracheal cannula. Emergency treatment included overinflation of the tracheostomy cuff and surgery via sternotomy, with occlusion of the tracheostomy and the tracheo arterial fistula and reimplantation of the brachiocephalic artery via a vascular prosthesis. This life-threatening complication is due to close anatomic relationships between the trachea and the brachiocephalic artery. In our case, the tip of the cannula eroded the anterior trachea wall resulting in a tracheo arterial fistula with massive haemorrhage. Warning symptoms such as pulsations of the cannula and aspiration of blood must be recognised without delay and followed by an adequate pre-established management. PMID- 7486302 TI - [Paraplegia after aorto-bifemoral prosthesis: systematized dysautonomic symptoms should evoke spinal cord damage]. AB - We report a case of paraplegia following aorto-bifemoral by-pass surgery for infrarenal aortic occlusive disease. The first symptoms of spinal cord damage were systematized dysautonomic symptoms including mottled skin and sudation. Such symptoms, occurring during recovery from anaesthesia are an indication for checking the status of the spinal cord. PMID- 7486303 TI - [Postoperative disorders of hemostasis caused by antifactor V circulating anticoagulant]. AB - The case of a 34-year-old woman who experienced a biological haemostasis disorder, without clinical manifestations, is reported. The disorder was caused by a factor V inhibitor induced by a prolonged administration of antibiotics required by a postoperative biliary peritonitis. Therapeutic strategies in patients with an acquired factor V inhibitor are reviewed. PMID- 7486305 TI - [Trans-thecal digital anesthesia in a case of section of the flexor tendon sheath]. PMID- 7486304 TI - [Locoregional anesthesia for endoscopic surgery of the carpal tunnel]. AB - Local anaesthesia for surgical endoscopic release of carpal ligament is obtained with a block of the median nerve associated with subcutaneous infiltration of the areas of entrance and exit of the endoscope. A palmar application of Emla cream makes the needle puncture painless. The accidental puncture of the median nerve, which occurs when the needle is inserted too rapidly, is the only potential complication. It is easily prevented by pushing the needle gently forward and orientating the bevel parallel to the axis of the nerve. The use of a neurostimulator and needle with a blunt tip can be an alternative. We used this technique in more than 1,500 patients. Only one accidental puncture of the nerve due to a technical error occurred. PMID- 7486306 TI - [Sluder tonsillectomy in children: can intratracheal intubation be discussed?]. PMID- 7486307 TI - [A new technique for manual ventilation assistance]. PMID- 7486310 TI - [Use of red blood cells to compensate for surgical blood loss in adults. Consensus development conference. Paris, 17 December 1993]. PMID- 7486309 TI - [Deep accidental hypothermia]. PMID- 7486311 TI - [Relations between body oxygen requirement, transport, consumption and tissue extraction]. PMID- 7486308 TI - [A cause for error in automatic detection of halogenated agents: alcoholic intoxication]. PMID- 7486312 TI - [Perspectives for the use of oxygen carriers as erythrocyte substitutes in surgery]. PMID- 7486313 TI - [Which lower value of hematocrit or hemoglobin should guide the transfusion of erythrocyte concentrates during and after extracorporeal circulation?]. PMID- 7486315 TI - [Which marker of erythrocyte volume should be used: hemoglobin, hematocrit, laboratory or medical test, or isotopic determination of erythrocyte volume?]. PMID- 7486316 TI - [When, in which patients and for what surgical procedures should programmed autologous transfusion be considered? Which quantitative benefit should be expected?]. PMID- 7486314 TI - [Which value of hematocrit and/or hemoglobin concentration should indicate the need for erythrocyte transfusion in a pregnant woman during and after cesarean section?]. PMID- 7486317 TI - [Programmed autologous transfusion. Logistical problems and management of patients with viral markers]. PMID- 7486318 TI - [Preoperative autologous erythrocytapheresis]. PMID- 7486319 TI - [Which technique should be chosen to reuse blood lost intraoperatively? Does the type of surgery constitute any contraindication for reutilization (cancer, infection)?]. PMID- 7486320 TI - [Techniques of salvage of blood lost during the postoperative period]. PMID- 7486321 TI - [Indications of phenotyped, leukocyte-depleted, CMV negative or irradiated homologous erythrocyte concentrates in adult surgery]. PMID- 7486322 TI - [Are there any limits to hemodilution?]. PMID- 7486324 TI - [Perioperative artificial nutrition in planned surgery in adults. Consensus development conference. Saint-Mande, France, 16 December 1994]. PMID- 7486323 TI - [Role of recombinant erythropoietin during the preoperative period]. PMID- 7486325 TI - [Role of new nitrogen substrates during peri-operative artificial nutrition in adults]. AB - Some amino acids and their derivatives, such as arginine (ARG), glutamine (GLN) in free form or as dipeptides, and ornithine-ketoglutarate (OKG), have specific pharmacological properties concerning namely immunomodulation, control of protein turn-over, maintenance of gut trophicity. In the context of the postoperative nutrition in the adult, supplementation of enteral nutrition with ARG and of parental nutrition with GLN and OKG has improved nutritional and biochemical markers such as nitrogen balance, muscle protein synthesis and glutamine content. However, only few studies have tried to demonstrate a clinical benefit with such a supplementation. At present only the beneficial effect of OKG on postoperative wound healing has been recognized. PMID- 7486326 TI - [Role of hepatocellular insufficiency and kidney failure on nutritional management]. AB - Chronic liver disease as well as chronic renal failure are responsible for abnormal nutrient metabolism and high rates of undernutrition. Although surgery is frequently required in such patients, the perioperative nutritional management has not yet been extensively studied in these conditions. During chronic liver disease, preoperative nutritional status and postoperative outcome are correlated. However, nutritional status can only be considered as one out of several factors of the prognosis, including the grade of liver insufficiency and the type of surgical procedure. Thus, it is difficult to evaluate the real influence of undernutrition on postoperative outcome. Similarly, the usefulness of preoperative nutrition is not definitely demonstrated in this condition. The nutritional requirements of patients with liver cirrhosis are estimated to be protein 1g and 30-35 kcal.kg-1.d-1. The duration of nutritional supplementation before surgery may not exceed 10 days. Postoperative parenteral nutrition seems to be well tolerated during chronic liver disease. Although some data in the literature suggest that it may be of interest after liver transplantation, the efficacy of postoperative parenteral nutrition needs to be proven in larger series. In chronically uraemic patients, the effects of undernutrition on postoperative morbidity and mortality have been poorly studied. Infectious complications after renal transplantation are favoured by several factors, including immunosuppressive therapy and malnutrition. The efficacy as well as the tolerance of perioperative nutritional support in patients with chronic renal failure are poorly known. In haemodialysis patients, candidates for renal graft, the nutritional status is usually well preserved when the recommended nutritional needs are provided: proteins 1.2-1.4 g and 35-40 kcal.kg-1.d-1. PMID- 7486328 TI - [Pre- and post-operative enteral nutrition in elective surgery in adults. Techniques, advantages and adverse effects]. AB - This review examined the techniques for enteral nutrition (routes and methods of administration and choice of enteral diets) in the perioperative period of elective surgery in adults. Only few controlled studies have assessed the efficiency of techniques according to the indication of enteral feeding (pre or postoperative period, digestive or non-digestive surgery). The nasogastric tube remains the most appropriate method to deliver short-term enteral feeding during the preoperative period. Today percutaneous endoscopic gastrotomy is the preferred method of gastrostomy. It is indicated in long term enteral nutrition, for example in the perioperative period of cancer surgery of head and neck. In the postoperative period of digestive surgery, intrajejunal feeding is usually delivered by a surgical jejunostomy or by a nasojejunal tube. Controlled studies comparing these two methods are still lacking. An important limitation of intrajejunal feeding is the poor tolerance of enteral diet during the first postoperative days after major upper abdominal surgery. The choice of enteral diet in the perioperative period remains controversial. There is no clinical evidence to support the hypothesis that the use of pre-digested diets in jejunostomy feeding has a better nutritional benefit and a better tolerance than polymeric diets. Small peptides offer some metabolic advantages, however, the clinical superiority of these nutrients over polymeric diets is not definitively proven. Continuous administration of enteral nutrition is usually required in case of jejunal feeding. In the other cases, some data suggest that enteral nutrition is more efficient using an intermittent feeding regimen. PMID- 7486327 TI - [Artificial nutrition in patients with chronic respiratory insufficiency undergoing surgery]. AB - Denutrition is a common finding in patients with chronic respiratory failure (CRF). Preoperative denutrition increases the risk of nosocomial pneumonia and difficulties in weaning from mechanical ventilation. A preoperative nutritional support may have beneficial effects on respiratory muscles performance. However, prospective studies need to be carried out in patients with CRF to substantiate this hypothesis. Postoperative nutritional support is indicated if weaning from the ventilator is expected to require more than several days, in order to preserve the diaphragmatic function. Lipid-enriched nutrition may have a beneficial effect, when energy supply is high, as the resulting decrease in CO2 production may facilitate the weaning from the ventilator. A beneficial effect of branched-chain amino acid-enriched solutions has not been demonstrated in patients with CRF. PMID- 7486329 TI - [Respective indications of enteral or parenteral nutrition during pre- and post operative periods]. AB - Denutrition is often associated with poor postoperative outcome. However, a large body of evidence, from studies comparing perioperative parenteral (PN) or enteral (EN) nutrition to the absence of perioperative nutrition, suggests that perioperative nutritional support provides significant improvements in both nutritional status and postoperative clinical outcome in selected patients who are or will become malnourished. The aim of this study was to select and review all relevant articles comparing perioperative parenteral and enteral nutritional support, either in terms of clinical outcome, or risks and costs, or in pathophysiological terms. Twelve clinical reports were reviewed. All contained methodological flaws, mainly type II statistical error due to an insufficient number of patients, inaccurate primary diagnosis, absence of blinding, and lack of objective criteria of judgement. These concerns warrant caution in interpreting the results. Moderately strong (grade B) recommendations can only be drawn from these studies: PN (compared to early EN) is associated with a higher rate of sepsis in patients following abdominal trauma; EN is as efficient as PN in patients following surgery; EN is safe and cheaper than PN. PN formulae lack many important nutrients (glutamine, arginine, cysteine, peptides, fibers, n-3 polyunsaturated fatty acids, and nucleotides). Many experimental (animal) and some clinical (in non surgical patients) studies showed that PN (compared to EN) induces gut mucosal atrophy, liver dysfunction, gut bacterial translocation and immune dysfunction. The final aim of PN and EN would therefore strikingly differ. The qualitatively imperfect PN would only supply the fasting patient with quantitative amounts of calories and proteins. Due to initially limited digestive tolerance, EN provides less nutrition than PN does, but would finally lead to the same or even better outcome, due to its ability to counteract stress induced gut and immune dysfunction. Current evidence therefore suggests that early EN is superior to PN in trauma patients, and not different from but cheaper (and therefore more cost-effective) than PN in surgical patients. Further controlled, randomised, and blinded studies including sufficient sizes of groups are required, especially in the surgical setting, to address a large number of still unanswered questions. PMID- 7486330 TI - [How to assess preoperative nutritional status?]. AB - The assessment of nutritional status of patients should rely on sensitive and specific evaluation tools. However a universally recognized criterion of severity of denutrition is still lacking. The most important clinical indicator is the weight. The speed of weight loss is an important point to consider: a loss of 2% within one week is equivalent to 5% in one month and 10% in six months. Other anthropometric indices such as the triceps skin fold and the mid arm muscle circumference have not shown, when considered alone, to increase the risk carried by denutrition. Therefore other clinical indicators have to be considered, the association of which is suggestive of malnutrition, but none when considered alone is neither specific nor pathognomonic. Food intake is quantified by questioning. Among the biological markers representative of the pool of circulating visceral proteins is albumin. Numerous studies have demonstrated a morbidity and mortality increase when the plasmatic albumin concentration is less than 30 g.L-1. Other proteins with a shorter half-life, such as transferin, prealbumin and retinal binding protein have been recommended. The determination of muscular mass is also a tool for nutritional assessment. The muscular function and immunologic tests such as the lymphocyte count and cutaneous reactivity to various antigens have been assessed as denutrition indices. The determination of body mass composition by bioelectrical impedance could become the technique of choice for nutritional assessment. However its use for peri-operative evaluation has not been validated prospectively. Because of the lack of reliability of markers when considered alone, several authors produced multivariate nutritional indices.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7486332 TI - [Role of preoperative nutritional status on postoperative morbidity]. AB - Correlations between preoperative nutritional status on the one hand and postoperative morbidity and mortality on the other hand were reviewed in articles from literature. The inclusion criteria for analysis were the following: large series, well defined nutritional status, major surgery, objective evaluation of postoperative complications and relevant statistical analysis. This study considered factors supposed to alter this correlation, namely the type of primary disease, intensity of denutrition, obesity, site and grade of cancer and age. There was a highly significant correlation between preoperative denutrition and postoperative morbidity and mortality. This correlation was existing whether operation was performed on the gastrointestinal tract or not, whether the primary disease was a cancer or not. The more important the denutrition, the more frequent the postoperative complications and deaths. Morbidity and mortality rates were linearly correlated to denutrition. After head and neck surgery for malignant diseases, malnourished elderly patients had poor prognosis. Obesity had the same prognostic value as denutrition. PMID- 7486331 TI - [How to assess and monitor postoperative artificial nutrition?]. AB - Quantitative and qualitative nutritional requirements depend on the level of energetic expenses. Various formulas, especially the tables by Harris and Benedict allow the estimation of the level of energetic expenses with an approximation of 14%. Corrective factors permit an adjustment of the figures, according to the level of body aggression. In complex cases, indirect calorimetry allows a more accurate appraisal of energetic expenses. This technique provides also indications on the utilisation of each substrate and allows therefore to determine the optimal carbohydrate-lipid ratio for each patient. The assessment of the direct benefit of artificial nutritional support relies on anthropometric techniques and at present on body composition appraisal by determination of its impedance. The changes in muscular strength are difficult to assess. Moreover the time course of body weight is not specific for nutritional status. Therefore other biological indicators such as the nitrogen balance, the concentration of plasma proteins and albumin are more often assessed; proteins with a short half life depend on the body aggression level. The potassium balance, which is easy to obtain in clinical practice, is a relevant indicator for nitrogen balance and protein synthesis. Clinical monitoring includes the checking of hydratation and its impact on the circulatory, respiratory and renal functions. The tolerance of enteral nutrition is appraised by the quality of gastrointestinal function. Biological monitoring includes the electrolyte balance and various variables of carbohydrate, lipidic and proteic metabolisms. It allows to check the absence of hyperglycaemia, hyperlipidaemia and cholestasis. The daily checking of catheters is part of the monitoring of nutritional support.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7486333 TI - [Consequences of surgery on nutritional status]. AB - Trauma and surgery induce extensive physiological changes, commonly denominated the acute phase reaction (APR). This APR is activated by various kinds of stimuli, namely nociceptive stimulations, tissue injury, tissue ischaemia and reperfusion as well as by haemodynamic disturbances which occur commonly in such patients. APR is mainly characterised by the release of counter-regulatory hormones, complex metabolic changes and by the hepatic synthesis of numerous acute phase factors (C-reactive protein, haptoglobin, complement protein, etc). In addition, fever is typically present and there is a resistance to the nutritional support. The intensity of APR is variable, according to the extent of surgery, the subsequent course, occurrence of complications and to various factors related to the patient and its treatment. In patients with non complicated surgery and low or moderate severity trauma, the metabolic changes are minor and self-limited. In such a condition, there is no need for nutritional support. Conservely, in patients with complicated surgery or major trauma, there is an extensive APR, which can be very prolonged. This results in important and sustained metabolic changes, leading to extensive catabolism and progressive loss of body cell mass. The latter is amplified by the decreased body ability to adapt to starvation and by the resistance to the nutritional support that typically occur in complicated postoperative and trauma patients. Total parenteral nutrition does not prevent from metabolic changes occurring in surgical patients. By contrast, several experimental and human studies have shown that early enteral nutrition may alleviate both the endocrine and metabolic responses in such conditions. Regional anaesthesia, particularly by the epidural route, may also decrease but not abolish the extent of APR. PMID- 7486334 TI - [In which patients has the efficacy of preoperative artificial nutrition be proved?]. AB - Today a consensus on the value of peroperative nutritional support is not existing, despite a recognized correlation between the stage of denutrition and the postoperative mortality and morbidity rates. This review of 11 prospective randomized studies, comparing undernourished patients with a major malignant or non-malignant surgical illness, according to the use or not of preoperative nutritional support, showed a wide discrepancy between the techniques of denutrition assessment, the underlying surgical disease, the tumor extension, the extend of surgery, the level and duration of nutritional support between the studies. However, among them, those without major bias show that a preoperative nutritional support of at least seven days, decreases significantly the rate of postoperative complications, especially the infection, in the most undernourished patients. PMID- 7486335 TI - [Recommendations of the jury. Peri-operative artificial nutrition]. PMID- 7486336 TI - [In which patients and for which procedures has the efficacy of postoperative artificial nutrition be proven?]. AB - This study reviewed 19 prospective studies for the incidence of postoperative nutritional support on outcome in elective surgery. It compared enteral and parenteral nutrition initiated no more than three days preoperatively or postoperatively and prolonged maximally for one month, to a simple infusion of glucose or saline. As all studies had methodological weaknesses, concerning mainly the sample size, it is impossible to propose relevant recommendations. Nevertheless, among the 11 studies on total parenteral nutritional support (nitrogen and caloric supply with carbohydrates and/or lipids), three of them produced valuable results. As the available data do not show any beneficial effect, a routine postoperative nutritional support cannot be recommended, even in patients at high risk of postoperative complications. However for the latter a possible benefit cannot be totally excluded in some of them. When an alimentation per mouth cannot be started during the 8 to 10 days after surgery, an artificial nutritional support becomes mandatory. PMID- 7486338 TI - [Caloric and nitrogen intake during pre- and post-operative periods. method and duration]. AB - Preoperative nutritional support is analysed from 7 prospective studies. Clinical benefits from nutrient intake and duration are not demonstrable. In one study, preoperative long-chain triglycerides infusions are associated with more postoperative complications. Postoperative nutrition is analysed from 20 articles. No one considers the clinical benefit with regard to quantitative and qualitative intakes. PMID- 7486337 TI - [Effect of postoperative complications on nutritional status: therapeutic consequences]. AB - The occurrence of a postoperative complication represents an additional stress factor for patients and leads in many cases rapidly to a malnutrition status. Thus a nutritional support is required as soon as the foreseeable duration of starvation has a longer duration than one week. Considering its lower risk of septic complications and lower cost, enteral feeding should be initiated as soon as possible. Appraisal of caloric needs with standard formulas often leads to inappropriate nutritional management. Therefore the requirements should be assessed by indirect calorimetry if available. Nutritional support is a part of the management of a postoperative septic patient. It must be initiated when initial phase of haemodynamic instability is amended. Branched chain amino acids, medium chain triglycerides and other specific nutrients have failed to demonstrate a real clinical beneficial effect. In case of acute respiratory failure, nutritional support must be cautious with regard to caloric load, as carbohydrates may increase CO2 production and lipids may worsen hypoxaemia. In case of postoperative acute renal failure, nutritional management is facilitated by continuous haemofiltration techniques allowing an unlimited nutrient intake. Solutions containing only essential amino acids are not recommended. During severe acute pancreatitis, enteral feeding is indicated when ileus does not permit the use of the intestinal tract. Jejunal access must be preferred to stomach or duodenum. Lipid emulsions can be used safely if serum triglyceride concentrations remain below 4 g.L-1 during infusion and below 2 g.L-1 between infusions. PMID- 7486340 TI - [Role of anabolic hormonal factors in surgery in adults]. AB - Hormonal agents with anabolic effect are virtually able to limit or even to counteract the adverse effects of stress and to act in synergy with nutritional support. Hormones with a high theoretic anabolic power include recombinant human growth hormone (rGH), insulin, insulin-like growth factor and testosterone. Other molecules favour anabolism in modifying the catabolic effect of diseases (anti cytokines, beta-adrenoceptor agonists), in promoting intestinal absorption of substrates or in stimulating epithelial growth (EGF). The results of preliminary studies cannot be transferred into routine clinical practice. The rGH favours the preservation of body mass integrity and various functions. Therefore it could improve postoperative outcome and decrease the duration of hospital stay. A study in severely burnt children showed the metabolic benefits of rGH as well as a shortening of hospital stay because of a decrease of cicatrisation delay after skin grafting. Finally it demonstrated that rGH allows major financial savings in severe burns. The IGF-1 stimulates host defences and decreases the rate of wound infections. The rGH potentiates nitrogen sparing effect of IGF-1 and decreases hypoglycaemia induced by IGF-1. However a study on patients in stress is yet lacking. A combination of growth factors could be virtually efficient, however the cost seems exorbitant. An unquestionable evidence of their clinical efficiency is required before a clinical use in routine can be considered. The postoperative protein catabolism could be decreased by nandrolone decanoate or its derivatives. On the other hand, it has not been proven that protein synthesis, the muscle function or clinical prognosis can be improved by such a treatment.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7486339 TI - [Role of trace elements and vitamins in peri-operative nutrition]. AB - The micronutriment requirements, whether trace elements (inorganic) or vitamins (organic), are tightly linked to the carbohydrate, lipid and protein metabolism, since they are involved in all metabolic pathways as cofactors. The micronutriments also have major immunological, endocrinological and antioxydant functions. Especially in the surgical patient, individual requirements may vary considerably and will be particularly increased in case of prior deficiency, anabolic states, or increased losses (burns, diarrhoea, gastric aspiration, intestinal fistulae, alcoholism, use of renal replacement techniques). In some of these settings, the micronutriment requirements will be independent from the macronutriments: this has been demonstrated for burns and intestinal fistulae. In the case of depletion prior to surgery, an isolated supplementation may be required without starting a proper nutrition. In general, micronutriment supplements will have to be started upon initiation of any artificial nutrition. After elective surgery and in absence of specific losses, the micronutriment requirements will be linked to the metabolic state of the patient and to the energy-protein intakes. This is most striking for the vitamin B group, where the requirements are indicated in mg per 1000 kcal. Vitamins A and E are also at risk in the surgical patient. Recommended micronutriment supplements have been revised in 1994. Some trace element deficiencies (Se, Cr, Mo) can initiate very serious complications and will require special caution in the perioperative period. Other deficiencies (Cu, Zn) result in more slowly evolving clinical pictures, with lesser life-threatening potential, resulting in infections and prolonged wound healing. In such cases, multi-elementary supplements are inadequate, and single element solutions supplements are required. All the micronutriments are characterized by a dose-response curve. The quantity avoiding biochemical dysfunctioning in human pathological situations has not yet been established, and it is unsatisfactory to merely compensate for the losses. This notion of biochemical dysfunctioning phase preceeding the clinical deficiency syndrome is in investigation for many nutriments, especially as the importance of some micronutriments, such as Se and vitamin E, in maintaining antioxidant defences is clearly established. The potential for preventing free radical induced overproduction of cytokines by means of nutritional strategy and enhanced antioxidant defences clearly exists, and is only at an early phase of investigation in patients. The future will be marked by the development of nutritional pharmacology based on pathology-specific micronutriment supplements. PMID- 7486341 TI - Starkfest--'94. 1st International Conference on Vision and Movement in Man and Machines. June 24-26, 1994, Berkeley, California. Proceedings. PMID- 7486342 TI - Lawrence Stark, M.D., biomedical engineer. PMID- 7486343 TI - New models of the oculomotor mechanics based on data obtained with chronic muscle force transducers. AB - Several phenomenological models of the oculomotor mechanics that produce saccadic eye movements have been developed. These models have been based on measurements of macroscopic muscle and orbital tissue properties and measurements of eye kinematics during saccades. We recorded the forces generated by the medial and lateral recti during saccades in an alert, behaving monkey using chronically implanted force transducers. With this new data, we tested the ability of the classic saccade models to generate realistic muscle force profiles. Errors in the predictions of the classic saccade models led to a reexamination of the current models of extraocular muscle. Both a phenomenological, Hill-type muscle model and an approximation to Huxley's molecular level muscle model based on the cross bridge mechanism of contraction (distribution moment model) were derived and studied for monkey extraocular muscle. Simulations of the distribution moment model led to insights suggesting (i) specific modifications in the lumped force/velocity relationship in the Hill-type model that resulted in this type of phenomenological model being able to generate realistic dynamics in extraocular muscle during saccades; (ii) the distribution of activity in the different fiber types in extraocular muscle may be central to the characteristics exhibited by the muscle during saccades; (iii) the transient properties of lengthening muscle such as yielding are not significant during saccades; and (iv) the series elastic component in active muscle may be predominantly generated by the elastic properties of the cross-bridges. PMID- 7486345 TI - Estimation of autonomic nervous activity using the inverse dynamic model of the pupil muscle plant. AB - In order to elucidate the mechanism of the pupillary control system, the internal property of the pupillary muscle plant, as well as the autonomic nervous input to the muscle plant, must be analyzed. In this study, we approach the problem first by constructing a new homeomorphic biomechanical model for the human pupillary muscle plant (forward dynamic model). We showed that the model is able not only to reproduce various experimental results that exhibit various nonlinearities but also to explain how such nonlinear responses are generated in terms of the internal property of the model. Then, we contrive a possible method to estimate the autonomic nervous input to the muscle plant. This method utilizes the inverse dynamic model of the pupillary muscle plant so that the autonomic nervous input can be estimated from the pupillary response. We applied this method to the experimental step responses, and showed that the estimated neural input indicates characteristics quite similar to the results of the physiological experiment. Last, we discuss the origin of the pupillary escape and capture as well as the sustained and transient components of the pupillary response, based on the analysis of the forward and/or inverse dynamic model. PMID- 7486344 TI - An improved muscle-reflex actuator for use in large-scale neuro-musculoskeletal models. AB - This paper extends the systematic approach described in Winters and Stark (62) for developing muscle models. The underlying motivation is our finding that for larger scale shoulder and head-neck postural systems to be mechanically stable, open-loop muscle properties are often not sufficient. There are three primary contributions. First, the previous muscle mechanical model structure and parameter estimation process of (62) is updated to reflect recent experimental findings. Second, an intrafusal (IF) muscle model is developed that includes a gamma static motoneuron (MN) drive, a Hill muscle model, and a muscle spindle sensor across the IF series element; this provides a more appropriate muscle spindle output signal, especially for studies of posture. Third, the conceptual cut between the neuro-control input and the actuator is raised from just below the MN summing junction to a higher location, allowing a "muscle-reflex actuator" to be defined that satisfies the formal theoretical requirement for possessing passive spring-like behavior when the neurocontrol input is constant, alpha-gamma MN coactivation is assumed, and three types of intrinsic autogenic reflex responses (spindle, Golgi tendon organ, Rhenshaw cell) are developed. Default feedback gains are set based on the criteria that inherent feedback should not sculpt the feedforward excitation drive by more than +/- 10% of maximum. This new actuator model only mildly affects voluntary goal-directed dynamic performance, but enhances spring-like performance around the postural equilibrium state, in line with available animal and human studies and with several theories on postural regulation. PMID- 7486346 TI - "Adequate control theory" for human single-joint elbow flexion on two tasks. AB - The control of distance and speed during single-joint human elbow flexion is accomplished by different models of activating the motoneuron pools. Distance is controlled by modulating the duration of activation, while speed is controlled by modulating the intensity. The experiments reported on here compare movements of different distances under two sets of instructions: subjects moved either as fast and accurately as possible or in a specified time. The first task showed duration modulation, whereas the second, which required simultaneous control of distance and speed, showed both duration and intensity modulation. These results are interpreted in the context of a model for motor control, predicated on the existence of movement plans that use prior knowledge of the dynamics of the movement task to generate muscle activation patterns that produce joint torques. These plans use a simple algorithm based upon parameters of the task such as distance, load, and speed. From this plan, the kinematic trajectory emerges. PMID- 7486347 TI - The anthroform biorobotic arm: a system for the study of spinal circuits. AB - This paper reports the design, construction, and testing of a replica of the human arm, which aims to be dynamically as well as kinematically accurate. The arm model is actuated with McKibben pneumatic artificial muscles, and controlled by a special purpose digital signal processing system designed to simulate spinal neural networks in real time. An artificial muscle spindle has also been designed and tested. Design and test data are reviewed, and the paper describes how we hope to use the system to improve our understanding of the reflexive control of human movement and posture. PMID- 7486350 TI - Two methods for recommending bat weights. AB - Baseball players swung very light and very heavy bats through our instrument and the speed of the bat was recorded. These data were used to make mathematical models for each person. Then these models were coupled with equations of physics for bat-ball collisions to compute the Ideal Bat Weight for each individual. However, these calculations required the use of a sophisticated instrument that is not conveniently available to most people. So, we tried to find items in our database that correlated with Ideal Bat Weight. However, because many cells in the database were empty, we could not use traditional statistical techniques or even neural networks. Therefore, three new methods were used to estimate the missing data: (i) a neural network was trained using subjects that had no empty cells, then that neural network was used to predict the missing data, (ii) the data patching facility of a commercial software package was used, and (iii) the empty cells were filled with random numbers. Then, using these fully populated databases, several simple models were derived for recommending bat weights. PMID- 7486348 TI - A review of the role of efference copy in sensory and oculomotor control systems. AB - Efference copy is an internal copy of a motor innervation. In the oculomotor system it provides the only extraretinal signal about eye position that is available without delay, and it is shown to be the most important extraretinal source of information for perceptual localization and motor activity. Efference copy accompanies all voluntary eye movements and some involuntary ones, including pursuits, saccades, and the fast phases of vestibular and optokinetic nystagmus. Not all eye movements are accompanied by an efference copy; its presence is determined by a movement's function, not it dynamics. Because the gain of the efference copy mechanism is less than 1, and it does not take account of oculomotor delays and kinematics, it is supplemented by other mechanisms in achieving space constancy. It functions differently for perception and for visually guided behavior. There is only one efference copy for both eyes, reflecting Hering's law, and it is subject to adaptation. PMID- 7486349 TI - Egocentric visual target position and velocity coding: role of ocular muscle proprioception. AB - Limited knowledge is available regarding the processes by which the brain codes the velocity of visual targets with respect to the observer. Two models have been previously proposed to describe the visual target localization mechanism. Both assume that the necessary information is derived from the coding of the position of the eye in the orbit, either through a copy of the muscular activation (out flow model) or through eye muscle proprioception (in flow model). Eye velocity coding might be derived from velocity sensitive ocular muscle proprioceptors or from position coding signals through differentiation. We used techniques based on manual pointing and manual tracking of visual target, combined with passive deviation of one covered eye, to demonstrate that ocular muscle proprioception is involved in (i) eye-in-head position coding, hence in target localization function; (ii) long-term maintenance of ocular alignment (phoria); and (iii) sensing of visual target velocity with respect to the head. These observations support other data now available, describing the processes by which the brain codes position and velocity of visual targets. Such findings might interest engineers in the field of robotics who are facing the problem of providing robots with the ability to sense object position and velocity in order to create an internal model of their working environment. PMID- 7486351 TI - Training in virtual and real environments. AB - Transfer of training between real and virtual environments was examined using a pick-and place task with two different difficulty levels. The task was to minimize the time to move cans from one color coded location in the first row to the same color coded location in the back row and then to reverse the process. In the first task, the front and back disk colors were aligned, and in the second disk order, the front and back disk colors were randomly placed on the table. Subjects trained in one environment were then tested in the other and their performance compared with that of subjects being trained in that environment. Some virtual world-trained subjects showed small but significant improvement in performance compared with the untrained subjects for the real world task for both disk arrangements. The differences in performance between the two groups decreased with trial number until no difference was seen at the end of the sessions. None of the real world-trained subjects showed any significant improvement when performing the task in the virtual world compared with the untrained subjects. These results suggest that transfer-of-training from virtual to real world tasks can take place under certain conditions. PMID- 7486352 TI - Looking around: 35 years of oculomotor modeling. AB - Eye movements have attracted an unusually large number of researchers from many disparate fields, especially over the past 35 years. The lure of this system stemmed from its apparent simplicity of description, measurement, and analysis, as well as the promise of providing a "window in the mind." Investigators in areas ranging from biological control systems and neurological diagnosis to applications in advertising and flight simulation expected eye movements to provide clear indicators of what the sensory-motor system was accomplishing and what the brain found to be of interest. The parallels between compensatory eye movements and perception of spatial orientation have been a subject for active study in visual-vestibular interaction, where substantial knowledge has accumulated through experiments largely guided by the challenge of proving or disproving model predictions. Even though oculomotor control has arguably benefited more from systems theory than any other branch of motor control, many of the original goals remain largely unfulfilled. This paper considers some of the promising potential benefits of eye movement research and compares accomplishments with anticipated results. Four topics are considered in greater detail: (i) the definition of oculomotor system input and output, (ii) optimization of the eye movement system, (iii) the relationship between compensatory eye movements and spatial orientation through the "internal model," and (iv) the significance of eye movements as measured in (outer) space. PMID- 7486353 TI - Determination of the in situ forces and force distribution within the human anterior cruciate ligament. AB - The in situ forces and their distribution within the human anterior cruciate ligament (ACL) can clarify this ligament's role in the knee and help to resolve controversies regarding surgical treatment of ACL deficiency. We used a universal force-moment sensor (UFS) to determine the magnitude, direction, and point of application of the in situ forces in the ACL in intact human cadaveric knees. Unlike previous studies, this approach does not require surgical intervention, the attachment of mechanical devices to or near the ACL, or a priori assumptions about the direction of in situ force. Anterior tibial loads were applied to intact knees, which were limited to 1 degree of freedom at 30 degrees flexion. The in situ forces developed in the ACL were lower than the applied force for loads under 80 N, but larger for applied loads of more than 80 N. The direction of the force vector corresponded to that of the anteromedial (AM) portion of the ACL insertion on the tibial plateau. The point of force application was located in the posterior section of the anteromedial portion of the tibial insertion site. The anterior and posterior aspects of the anteromedial portion of the ACL supported 25% and 70% of the in situ force, respectively, with the remainder carried by the posterolateral portion. We believe that the data obtained with this new UFS methodology improves our understanding of the role of the ACL in knee function, and that this methodology can be easily extended to study the function of other ligaments. PMID- 7486354 TI - Yield of fluorescence from indocyanine green in plasma and flowing blood. AB - The purpose of this work was to obtain more quantitative knowledge about the yield of fluorescence from retinal vessels during indocyanine green angiography (ICG). The yield of fluorescence from blood was investigated for various shear rates, concentrations of ICG, and layer thicknesses. Measurements were performed in vitro on samples of human blood in a cone-plate shear chamber using frontal illumination as in scanning laser angiography. In blood and in plasma, the yield of fluorescence of ICG increased with concentration up to 0.05 and 0.1 mg/ml, respectively. At higher concentrations, the yield decreased for all layer thicknesses. For increasing layer thicknesses, both in plasma and in blood, the yield of ICG fluorescence increased nonlinearly for concentrations higher than 0.012 mg/ml. Saturation occurred for layers thicker than 200 microns in combination with ICG concentrations of 0.4 mg/ml and higher. Application of shear rates within the physiological range of the microcirculation (88/sec and 528/sec) increased the yield of fluorescence from the blood sample compared with stasis. The high transparency of blood for the excitation and emission light of ICG that was demonstrated will lead to superposition of fluorescence from superficial and deeper layers. This superposition precludes quantitative indocyanine angiography of ocular vessels. PMID- 7486356 TI - Evaluation of the dispersional analysis method for fractal time series. AB - Fractal signals can be characterized by their fractal dimension plus some measure of their variance at a given level of resolution. The Hurst exponent, H, is < 0.5 for rough anticorrelated series, > 0.5 for positively correlated series, and = 0.5 for random, white noise series. Several methods are available: dispersional analysis, Hurst rescaled range analysis, autocorrelation measures, and power special analysis. Short data sets are notoriously difficult to characterize; research to define the limitations of the various methods is incomplete. This numerical study of fractional Brownian noise focuses on determining the limitations of the dispersional analysis method, in particular, assessing the effects of signal length and of added noise on the estimate of the Hurst coefficient, H, (which ranges from 0 to 1 and is 2 - D, where D is the fractal dimension). There are three general conclusions: (i) pure fractal signals of length greater than 256 points give estimates of H that are biased but have standard deviations less than 0.1; (ii) the estimates of H tend to be biased toward H = 0.5 at both high H (> 0.8) and low H (< 0.5), and biases are greater for short time series than for long; and (iii) the addition of Gaussian noise (H = 0.5) degrades the signals: for those with negative correlation (H < 0.5) the degradation is great, the noise has only mild degrading effects on signals with H > 0.6, and the method is particularly robust for signals with high H and long series, where even 100% noise added has only a few percent effect on the estimate of H. Dispersional analysis can be regarded as a strong method for characterizing biological or natural time series, which generally show long-range positive correlation. PMID- 7486357 TI - An optimal constrained linear inverse method for magnetic source imaging. AB - Magnetic source imaging is the reconstruction of the current source distribution inside an inaccessible volume from magnetic field measurements made outside the volume. It is possible in many applications to estimate, from prior physiological and anatomical knowledge, the source positions, amplitudes, and correlations, as well as the noise amplitudes and correlations. The optimal constrained linear inverse method (OCLIM) uses this prior knowledge to obtain a minimum mean-square error estimate of the current distribution. OCLIM can be efficiently computed using the Cholesky decomposition, taking about a second on a workstation-class computer for a problem with 64 sources and 144 detectors. Any source and detector configuration is allowed as long as their positions are fixed a priori. Correlations among source and noise amplitudes are permitted. OCLIM reduces to the optimally weighted pseudoinverse method of Shim and Cho if the source amplitudes are independent and identically distributed and to the minimum-norm least-squares estimate in the limit of no measurement noise or no prior knowledge of the source amplitudes. In the general case, OCLIM has better mean-square error than either previous method. OCLIM appears well suited to magnetic imaging, since it exploits prior information, provides the minimum reconstruction error, and is inexpensive to compute. PMID- 7486355 TI - Characterization of the cellular microstructure of ocular lens using 2D power law analysis. AB - Power law analysis provides a quantitative method for characterization of spatial fluctuations in the cellular microstructure of the ocular lens. In the power law analysis, Fourier components of the spatial fluctuations are computed, and the relationship between the amplitude, A, and spatial frequency, f, of the components is defined by a power law function: [formula, see text]. The exponent of the function, beta, defines the scaling of the amplitude of the Fourier components as a function of spatial frequency. We performed two-dimensional power law analysis on electron micrographs of lens cells ranging from transparent to opaque. We identified two values of power law exponent, beta, for the spatial fluctuations of all lens cells, one for low- and a second for high-spatial frequencies. In the low-spatial frequency region, the value of beta was in the range of 0.53 to 1.33, for transparent and opaque cells. In the high-spatial frequency region, the value of beta increased from 2.78 for transparent lens cells to 3.60 for opaque lens cells. The power law analysis provides a new method for quantitative characterization of the spatial fluctuations in the microstructure of transparent and opaque lens cells. PMID- 7486358 TI - 1995 Annual fall meeting of the Biomedical Engineering Society. Boston, Massachusetts, October 6-8, 1995. Abstracts. PMID- 7486359 TI - Randomized, placebo-controlled evaluation of prochlorperazine versus metoclopramide for emergency department treatment of migraine headache. AB - STUDY OBJECTIVE: To determine the comparative efficacy of i.v. metoclopramide and prochlorperazine for the initial emergency department treatment of migraine headache. DESIGN: Prospective, randomized, double-blind, placebo-controlled trial. SETTING: Military community hospital ED with an annual census of 75,000. PARTICIPANTS: Seventy consenting adults from a convenience sample of patients presenting with migraine headache similar to that experienced in at least one prior episode. Exclusion criteria were pregnancy, fever, signs of meningismus, altered sensorium, drug or alcohol use, oxygen saturation less than 90%, recent trauma or seizure, "worst headache," abnormal blood pressure, recent (within 48 hours) use of metoclopramide or prochlorperazine, and allergy to metoclopramide or prochlorperazine. INTERVENTIONS: In a random manner, each subject received a 2 mL i.v. injection of identical-appearing fluid containing metoclopramide (10 mg), prochlorperazine (10 mg), or saline solution (placebo). No other analgesics or medications were administered during the initial study period; rescue agents were administered by the choice of the treating physician after all data were collected. MEASUREMENTS: Patients scored their nausea, pain, and sedation before receiving the 2-mL injections and at 30 minutes after injection. Ten-centimeter nonhatched visual analog scales were used for these measurements, with distance from the left end (zero) calculated for each use. Clinically important successful treatment was defined a priori as achievement of the following criteria: patient satisfaction and either a decrease of 50% or more in the 30-minute pain score (compared with the initial score) or an absolute pain score of 2.5 cm or less. Failure to achieve these criteria constituted treatment failure. Differences between groups were analyzed with the Kruskal-Wallis ANOVA and chi 2 tests. Data are reported as frequency percentages and median values, with a two-tailed P value of .05 or less considered significant. RESULTS: Nausea, pain, and sedation scores were similar in all three groups before therapy. Thirty minutes after treatment, pain scores differed among those treated with prochlorperazine (1.1 cm), with metoclopramide (3.9 cm), and with placebo (6.1 cm, P = .003). Clinical success occurred more commonly after treatment with prochlorperazine (82%) than after metoclopramide (46%) or placebo (29%, P = .03). However, metoclopramide and placebo scores did not differ (P = .14). Nausea tended to be improved after prochlorperazine, compared with metoclopramide or placebo, at 30 minutes (P = .64). Four patients (6%) returned to the ED for relapse of migraine headache within 24 hours (three in the placebo group and one in the metoclopramide group). CONCLUSION: i.v. prochlorperazine relieves the headache and tends to improve nausea better than metoclopramide in ED patients with acute migraine headache. PMID- 7486360 TI - Outpatient management of acute urticaria: the role of prednisone. AB - STUDY OBJECTIVE: To evaluate the efficacy of a 4-day "burst" course of prednisone added to standard treatment with H1 antihistamines for the management of acute urticaria in outpatients. DESIGN: Prospective, randomized, double-blinded, clinical trial. SETTING: Emergency department of an urban tertiary care teaching hospital. PARTICIPANTS: Adult patients with urticarial rash of no more than 24 hours' duration, regardless of cause. Patients were excluded if they manifested wheezing, stridor, or angioedema or if they had taken antihistamines or glucocorticoids within 5 days of arrival at the ED. Patients also were excluded if there was a history of diabetes or active peptic ulcer disease. INTERVENTIONS: All patients were asked to evaluate the severity of pruritus ("itch score") on a 10-cm visual analog scale. Patients were then given diphenhydramine, 50 mg intramuscularly, and discharged home on a regimen of hydroxyzine, 25 mg orally, every 4 to 8 hours for pruritus, plus either prednisone, 20 mg, or placebo orally every 12 hours for 4 days. Patients' conditions were reassessed clinically, with itch score calculated again 2 days later, and again at 5 days by telephone. RESULTS: Forty-three patients were enrolled; 24 received prednisone and 19 received placebo. The two groups had similar itch scores at enrollment (prednisone, 8.1 +/- 1.7; placebo, 7.4 +/- 2.1, P = .25 [ANOVA]), but at 2- and 5 day follow-up the prednisone group had significantly lower itch scores (1.3 +/- 1.3 and .0 +/- .0 versus 4.4 +/- 2.2 and 1.6 +/- 1.0, respectively; P < .0001 [ANCOVA] at each interval) and greater clinical improvement in rash. Response did not correlate with age, sex, or identification of an allergen. No adverse effects were noted in either group. CONCLUSION: The addition of a prednisone burst improves the symptomatic and clinical response of acute urticaria to antihistamines. Patients' conditions improved more quickly and more completely when prednisone was administered, without any apparent adverse effects. PMID- 7486362 TI - Is circadian variation in asthma severity relevant in the emergency department? AB - STUDY OBJECTIVE: Asthmatic patients have marked circadian variation in disease severity, with bronchospasm far worse between midnight and 8 AM than at other times of day. In this study we sought to determine whether circadian variation in asthma severity is clinically relevant for purposes of emergency management. DESIGN: Prospective, observational cohort study. SETTING: The emergency department of a large urban university hospital. PARTICIPANTS: Asthmatic patients 16 years and older who presented with the complaint of asthma exacerbation. RESULTS: Peak expiratory flow rate (PEFR), respiratory rate, and objective and subjective assessments of dyspnea severity were determined at the time of each patient's presentation to the ED and at the time of disposition decision. Time of symptom onset, duration of symptoms, response to therapy, and final disposition were recorded. Nocturnal presentation and onset were defined as arrival in the ED or onset of symptoms between midnight and 8 AM. We enrolled 188 asthmatic patients in the study; 52 (28%) presented nocturnally. Comparison of asthmatic patients who presented nocturnally and those who presented at other times revealed no differences in initial or final PEFR or respiratory rate, change in PEFR, subjective or objective dyspnea, or rate of hospitalization, although nocturnal patients did have a significantly shorter ED treatment time. Analysis by time of symptom onset also revealed no differences between nocturnally and nonnocturnally presenting subjects in disease severity or outcome, with the exception that those with nocturnal symptom onset had less subjective dyspnea than other asthmatic patients. Repeat analysis with different "nocturnal" time periods did not alter the results. CONCLUSION: In our study group, asthmatic patients who presented nocturnally to the ED did not differ from other asthmatic patients in disease severity and therefore do not appear to represent a unique population for purposes of ED management or disposition decisions. PMID- 7486361 TI - Treatment of acute bronchospasm with beta-adrenergic agonist aerosols delivered by a nasal bilevel positive airway pressure circuit. AB - STUDY OBJECTIVE: To determine whether beta-adrenergic agonist aerosols are more effective in treating acute bronchospasm if delivered by nasal bilevel positive airway pressure (BiPAP) than by a small-volume nebulizer (SVN). We hypothesized that BiPAP would reduce the work of breathing in asthmatic patients and thereby hasten improvement of bronchospasm from beta-agonist therapy. Previous trials with aerosols given by intermittent positive-pressure breathing were unrewarding, but BiPAP is unique in that inspiratory (IPAP) and expiratory (EPAP) support pressures may be set separately. DESIGN: Convenience-randomized prospective clinical study. SETTING: Emergency department of an urban tertiary care teaching hospital. PARTICIPANTS: Afebrile, wheezing patients between 18 and 40 years of age. INTERVENTIONS: Patients were randomly assigned to receive two doses of aerosolized albuterol (2.5 mg in 3 mL normal saline solution), 20 minutes apart, delivered by either SVN (n = 40) or BiPAP (n = 60) by nosemask or facemask (IPAP, 10 cm H2O; EPAP, 5 cm H2O). RESULTS: Peak expiratory flow rate (PEFR), arterial blood oxygen saturation (by pulse oximetry), and pulse and respiratory rates were measured at baseline and after each treatment. The two treatment groups had similar values for pulse oximetry, pulse rate, respiratory rate, and percent of predicted peak expiratory flow rate (%PPEFR) at entry, and all patients experienced similar changes in the first three of these variables through the course of treatment. BiPAP patients had a significantly greater increase in %PPEFR after each treatment (P = .0011) and from baseline to completion (P = .0013). Increase in absolute PEFR was greater in the BiPAP group (from 211 +/- 89 [mean +/- SD] to 357 +/- 108 L/minute for BiPAP, from 183 +/- 60 to 280 +/- 87 L/minute for SVN; P = .0001). CONCLUSION: In this population, response to initial ED management of bronchospasm, as measured by PEFR, was better with aerosols delivered by BiPAP than with those delivered by SVN. PMID- 7486363 TI - Pavement temperature and burns: streets of fire. AB - STUDY OBJECTIVE: To measure pavement temperatures over a 24-hour period to determine when patients are at risk for burns and to report cases of pavement burns with predisposing factors. DESIGN: Descriptive study of pavement temperatures and retrospective case series of 23 patients with pavement burns admitted to the Maricopa Medical Center during the years 1986 to 1992. PARTICIPANTS: Twenty-three patients with pavement burns serious enough for them to be admitted to the burn center. RESULTS: We measured the temperatures of asphalt, cement, and other outdoor materials hourly for one 24-hour period using a thermocouple thermometer. Asphalt pavement was hot enough to cause burns from 9 AM to 7 PM during the summer months. It was hot enough to cause a second-degree burn within 35 seconds from 10 AM to 5 PM. The group of burned patients could be divided into three categories: incapacitated, restrained, and sensory deficient. All burns involved less than 13% of the total body surface area. CONCLUSION: During summer days in the desert, pavement is often hot enough to cause burns and does so with regularity in the southwestern United States. No one should be allowed to remain in contact with hot pavement, even transiently. PMID- 7486364 TI - Effect of multiple-dose activated charcoal on the clearance of high-dose intravenous aspirin in a porcine model. AB - STUDY OBJECTIVE: To study the effect of multiple-dose activated charcoal (MDAC) on salicylate clearance in pigs given high-dose i.v. aspirin. DESIGN: In a crossover design, six fasted pigs received 300 mg/kg i.v. aspirin followed by no treatment or MDAC (1 g/kg hourly for 6 doses by gastrostomy). Serum salicylate samples were obtained every 30 minutes for 6 hours. RESULTS: The mean peak salicylate concentrations were 47.4 +/- 6.2 mg/dL and 48.4 +/- 3.9 mg/dL (P = .74), and the areas under the time-serum salicylate concentration curve over 6 hours were 171,000 +/- 24,000 mg.minute/L and 188,000 +/- 18,000 mg.minute/L for the control and treatment arms, respectively (P = .22). This study had a 90% power to detect a 30% difference between arms. CONCLUSION: MDAC does not enhance the clearance of salicylate after administration of high-dose i.v. aspirin. PMID- 7486365 TI - Comparison of tensile strength of cyanoacrylate tissue adhesive closure of lacerations versus suture closure. AB - STUDY OBJECTIVE: To compare the tensile strength of lacerations closed with cyanoacrylate tissue adhesive, with suture, and with a combination of tissue adhesive and suture. DESIGN: Randomized, controlled animal model study. SETTING: Animal care facility. PARTICIPANTS: Sixteen albino guinea pigs. INTERVENTIONS: Four incisions to deep fascia were created on the dorsum of each animal. Lacerations were closed with either tissue adhesive alone, tissue adhesive with subcutaneous suture, percutaneous suture alone, or percutaneous suture with subcutaneous suture. After 4 days, the breaking strengths of all wound closures were measured. RESULTS: Wound closures made with cyanoacrylate tissue adhesive alone exhibited significantly less tensile strength at 4 days than closures made by the other three methods. Wound closures made with combined percutaneous and subcutaneous sutures were significantly stronger than all other groups. CONCLUSION: Wound closures made with cyanoacrylate tissue adhesive alone have less tensile strength at 4 days than those made with tissue adhesive and subcutaneous suture or those closed by traditional suture methods. PMID- 7486366 TI - Comparison of ambulance dispatch protocols for nontraumatic abdominal pain. AB - STUDY OBJECTIVE: To compare rates of undertriage and overtriage of six ambulance dispatch protocols for the presenting complaint of nontraumatic abdominal pain, and to identify the optimal protocol. DESIGN: Retrospective prehospital and emergency department chart review to classify patients' conditions as "emergency" or "nonemergency." Utility analysis was used to identify the preferred protocol and monetary cost-effectiveness analysis to identify the least expensive protocol. SETTING: County emergency medical services (EMS) system with five receiving hospitals serving a mainly urban population of approximately 350,000. PARTICIPANTS: Records of 902 patients who called 911 for nontraumatic abdominal pain were reviewed; patients not transported were excluded. Twenty-seven county EMS medical directors completed questionnaires. RESULTS: Six ambulance dispatch protocols for nontraumatic abdominal pain were developed: indiscriminate dispatch, four selective protocols, and no-dispatch. A dichotomous classification system was derived prospectively from the prehospital and medical records of patients who had activated the EMS system before the study period to define "emergency" and "nonemergency" conditions associated with nontraumatic abdominal pain. Emergency criteria identified patients with conditions requiring medical treatment within 1 hour. Reviewers determined, for each patient, whether an ambulance would have been dispatched by each of the protocols. Undertriage and overtriage rates were calculated for each protocol. County EMS medical directors assigned utility values to four potential outcomes of ambulance dispatch by the direct scaling method. The outcomes comprised correct and incorrect decisions to dispatch ambulances to patients with and without emergencies. The protocols were compared by decision analysis. A cost analysis was also performed, using an estimated marginal cost per transport of $302. Sensitivity analysis demonstrated the effect of varying the cost of an undertriage error and the cost per response. Of the 788 patients included in the study, 7.8% had conditions defined as emergencies. The four selective ambulance dispatch protocols had overtriage rates ranging from 10% to 51% and undertriage rates of 4% to 7%. None of the protocols was proven superior on the basis of the medical directors' assignment of utility values. The marginal cost of dispatching advanced life support ambulances to all patients with this complaint was $3,838 per emergency. CONCLUSION: The majority of patients with nontraumatic abdominal pain who requested ambulance transport during the study period did not have conditions that were classified as emergencies. In the study model, if an undertriage error costs more than $3,674, indiscriminate ambulance dispatch is the least expensive protocol, and if an undertriage error costs less than $3,674, no ambulance dispatch is the least expensive strategy. PMID- 7486367 TI - Hawthorne effect: implications for prehospital research. AB - STUDY OBJECTIVE: The phenomenon of altered behavior or performance resulting from awareness of being a part of an experimental study has been termed the "Hawthorne effect." Prehospital studies generally involve paramedics or are designed to use data collected by paramedics. Our objective was to determine whether paramedic performance, as measured by frequency of documentation, can be modified by (1) written notification of the importance of documentation, (2) written notification of a research project involving paramedic documentation, or (3) written notification of a quality-improvement audit of paramedic documentation. DESIGN: Prospective, sequential intervention study with five study phases. SETTING: Urban, all-advanced life support public utility model emergency medical services system with 55,000 emergency calls per year. PARTICIPANTS: One hundred forty-five paramedics who completed all ambulance run reports from August 1992 to May 1993. RESULTS: A total of 30,828 run reports was entered into the study. Baseline undocumented parameters ranged from 3.7% to 6.5%. Compared with baseline, a memo to heighten awareness (phase 2) did not alter documentation (P > or = .08). A medication study memo (phase 3) improved medication documentation (P = .0005) and allergies documentation (P = .037). A quality-improvement audit memo (phase 4) improved documentation of all parameters (P < or = .001). CONCLUSION: The Hawthorne effect occurs in prehospital research. It does not require direct observation, nor does it require direct feedback. However, it may require a perceived demand for performance. The Hawthorne effect must be considered in the design of prehospital studies and interpretation of data collected by paramedics. PMID- 7486368 TI - Evaluation of a prehospital chest pain protocol. AB - STUDY OBJECTIVES: To evaluate the diagnostic accuracy and outcomes for patients treated by use of a prehospital chest pain protocol (CPP). DESIGN: Consecutive case series for 1 year (1993) of prehospital nontrauma advanced life support (ALS) cases including hospital outcomes. SETTING: Nonurban two-tiered emergency medical services system. PARTICIPANTS: Patients treated under the prehospital CPP or with hospital diagnoses of ischemic heart disease (IHD; ICD-9 between 410 and 414). Patients with cardiac arrest or dysrhythmias were excluded. INTERVENTIONS: Patients were given, by standing orders, ECG monitoring, i.v. access, and sublingual nitroglycerin. Further therapy was guided by on-line medical direction. RESULTS: Of 3,122 ALS nontrauma patients, 620 (20%) were treated with the CPP. All patients underwent ECG monitoring, i.v. access was started in 83%, and 61% received nitroglycerin. Only 55% of patients completed the entire CPP; patients who failed to complete the CPP had the same prevalence of IHD as those who completed it. When compared with hospital diagnosis of IHD, the CPP had a sensitivity of 69% (95% confidence interval [CI], 64% to 74%), a specificity of 87% (95% CI, 86% to 88%), and a positive predictive value of 42%. The positive likelihood ratio of CPP for IHD was 5.31, and the negative likelihood ratio was .36. The hospital mortality rate for all patients was 2.2%; for those with IHD, it was 1.6%. CONCLUSION: This prehospital ALS CPP had good diagnostic accuracy, but only half of patients completed it, and the hospital mortality rate was low. These data challenge the efficacy of the CPP. PMID- 7486373 TI - Ventricular fibrillation, automatic external defibrillators, and the United States Food and Drug Administration: confrontation without comprehension. AB - More people die in the United States each day of potentially reversible VF than of any other cause of death, reversible or not. Early defibrillation is the definitive treatment. Automated external defibrillation is a proven technology now confirmed to have saved thousands of lives. As with all medical devices and technology, perfection is not possible. Some problems, such as those represented by the two cases discussed in this article, are inevitable and acceptable and give little cause for alarm. One would not stop penicillin from being manufactured and distributed because of a sudden, unexpected allergic reaction in one patient (error of commission) or an unexpected resistant organism in another (error of omission). The FDA must understand that AEDs, even if they are imperfect, are not anywhere near as dangerous as no defibrillator at all. AEDs have finally allowed many EMS systems to achieve early defibrillation. Discontinuing use of AEDs or closing AED manufacturers could mean a significant number of lives lost unnecessarily. Therefore EMS agencies planning to implement early-defibrillation programs should continue with such plans. Why the agents of an important federal regulatory agency have singled out this technology for an intense review puzzles many observers in the medical-device field. Two meetings have been hosted by officials of the FDA to discuss the continuing concern the FDA officials have expressed over automated defibrillation technology. These meetings included representatives from the AHA, the American College of Cardiology, ACEP, defibrillator manufacturers, and other interested organizations. The FDA leadership has repeatedly focused on data acquired through the FDA Medical Device Reporting systems. Congress requires the FDA to investigate reports of problems with "critical medical devices." Because the indication for the use of a defibrillator is cardiac arrest, there will inevitably be a high association between defibrillator use and patient deaths. FDA personnel may view such reports of device problems in association with patient deaths as evidence that an intrinsically flawed technology has reached the marketplace without rigorous testing and evaluation. From the clinician's perspective, however, these reports represent a small numerator over a huge denominator of daily, lifesaving clinical use. The non-FDA participants at the two meetings have stated that the FDA complaints appear to be random and reveal a lack of understanding of AED technology.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 7486371 TI - Victim as offender in youth violence. AB - STUDY OBJECTIVE: To determine how often adolescent and young adult victims of assaultive injury are offenders in assaults and other crimes. DESIGN: Comparison of 10- to 24-year-old males treated in the accident and emergency department for assault-related injuries to similar-aged males treated in the same department for unintentional injuries. Police records were searched on both groups for warnings or convictions. SETTING: Accident and emergency department of the Cardiff Royal Infirmary. RESULTS: Assault patients were significantly more likely to be formally warned or convicted, and they had a higher mean number of warnings or convictions per 100 person-years of exposure, than other injury patients. These differences were most pronounced for the younger patients and for the year following the injury. CONCLUSION: These results suggest that many young male assault patients either have a history of criminal activity or develop criminal behavior subsequent to their assault and may benefit from appropriate intervention aimed at interrupting the cycle of crime and violence. PMID- 7486372 TI - Need and desire for preventive care measures in emergency department patients. AB - STUDY OBJECTIVE: To determine the need and desire for selected preventive care measures in an adult emergency department population, comparing patients with and without primary physicians. DESIGN: Written survey. SETTING: Urban university ED. PARTICIPANTS: English-speaking patients 18 years of age or older who did not arrive by ambulance, did not have a critical illness, and did not have a psychiatric complaint. RESULTS: The main outcome measures were past preventive care and desire to initiate preventive care measures as part of ED care. Nine hundred fifty-three surveys were distributed; 647 were completed and returned. Twenty-seven percent of patients knew their cholesterol level. Forty-three percent of men aged 40 years or older reported having had a prostate examination in the past year, and 39% of men aged 50 years or older reported having had an examination of stool for blood in the past year. Twenty-one percent of women reported taking calcium, and 67% of women aged 40 years or older had had a mammogram in the past 2 years. Sixty-three percent of patients had a primary physician; these patients were more likely to have received each of the preventive care measures studied (P < .025). Fifty-three percent of women not taking calcium requested information about osteoporosis prevention, and 30% requested prescriptions for calcium supplements. Sixty-two percent of women who did not have a current mammogram requested mammography information, and 60% requested referrals for mammography. Fifty-four percent of patients requested cholesterol and diet information. Patients requesting information, referrals, and prescriptions were given them. CONCLUSION: In a selected ED population, there was both need and desire for preventive health care measures to be initiated or provided as part of ED care, especially among patients who did not have primary physicians. PMID- 7486369 TI - Outbreak of Escherichia coli O157:H7 hemorrhagic colitis and hemolytic uremic syndrome: effect on use of a pediatric emergency department. AB - STUDY OBJECTIVE: To evaluate the effect of an outbreak of Escherichia coli O157:H7 colitis, and media coverage of the outbreak, on use of an emergency department. DESIGN: Review of pediatric ED use and charges for gastrointestinal illness during the epidemic and during a control period. SETTING: Pediatric ED in Seattle, Washington. PARTICIPANTS: All children seen in the ED with a gastrointestinal illness during the epidemic period (January and February 1993) and during a control period (January and February 1992). RESULTS: During the epidemic, 31 patients with E coli O157:H7 infection had 45 visits to the ED. The number of visits for gastrointestinal illness not caused by E coli O157:H7 was 103% higher in the epidemic period than in the control period (653 in 1992, 1,327 in 1993). The number of visits was closely associated with the number of newspaper stories about E coli O157:H7 illness (correlation coefficient, .88; P = .002). The increased number of evaluations for gastrointestinal illness not caused by E coli O157:H7 infection during the epidemic period was associated with an additional $101,193 in charges per month compared with the control period. There were no important differences in the evaluation of gastrointestinal illness between 1992 and 1993 except for an increase in the proportion of patients with stool cultures (13.1% versus 26.4%, P < .001). CONCLUSION: For every visit by a patient with E coli O157:H7 infection, there were 15 additional visits (above the baseline from the control period) by patients with other gastrointestinal illness. The true cost of this epidemic included not only the evaluation of patients with disease but also the evaluation of those who presented because they were worried they had the disease. The intensity of media coverage of a disaster may correlate with the number of visits to the ED, and understanding of this fact may help in disaster planning. PMID- 7486370 TI - Electrical injuries in a pediatric emergency department. AB - STUDY OBJECTIVE: To examine the spectrum of electrical injuries and develop guidelines for management. DESIGN: Retrospective review of charts compiled during a 6-year period (1988 through 1993). SETTING: Pediatric emergency department. PARTICIPANTS: Seventy-eight patients seen for electrical injuries. RESULTS: Fifty four percent of patients were boys, and the mean age of the patients was 5.3 years. Eighty-two percent sustained burns. We divided patients into those who were involved in major electrical current events (n = 8) (water contact and high voltage) and minor electrical current events (n = 70) (injury sustained while placing an object in an outlet or touching/plugging in a cord or during oral contact with a cord). Of the minor events, all burns (n = 61) involved less than 1% of body surface area. Eighteen patients sustained second-degree burns, and 19 sustained third-degree burns. Of the eight major-event patients, one had abnormal ECG/rhythm strip findings, two had abnormal urinalysis results, and six had abnormal levels of creatine phosphokinase. All eight were admitted. Of the 70 minor-event patients, 2 of 53 had abnormal ECG/rhythm strip findings, 1 of 48 had abnormal urinalysis results, and 2 of 40 had abnormal creatine phosphokinase levels. Thirty-six of the 70 minor-event patients were admitted. Patients involved in major events were more likely to undergo studies (P = .002), to have an abnormal result (P = .000008), and to be hospitalized (P = .008). In minor event patients, hospitalization was limited to observation and the fitting of oral appliances. CONCLUSION: Children involved in electrical events are usually exposed to low-voltage household current resulting in minor injury. Asymptomatic children with minor electrical injuries do not require laboratory evaluation or hospitalization. PMID- 7486374 TI - Defibrillation by basic emergency medical technicians: effect on survival. AB - Several studies have examined the effect of early defibrillation by basic EMTs on patient survival. Although the studies have a common theme of early basic EMT defibrillation, they are diverse in locations, devices, control groups, caregivers, and protocols. They provide a confusing array of information that is difficult to review, synthesize, and interpret. Metaanalysis allows data pooling of these primary studies to combine results and statistically compare the observed variation in study outcomes. The purpose of this metaanalysis was to examine the published studies of early basic EMT defibrillation to learn whether this treatment has an effect on survival of out-of-hospital cardiac arrest. Analysis of the 10 studies that met inclusion criteria showed that despite variations in design, the overall effect size for all the studies was .092, indicating a 9.2% increase in survival over what would have been expected had the EMT-Ds not intervened. PMID- 7486375 TI - Emergency medicine in Israel: state of the art. AB - Emergency medicine is in its infancy in Israel but is developing rapidly. Medical and government authorities such as the Israeli Medical Association and the Israeli Ministry of Health have already recognized the need for this field in Israel, although it remains for emergency medicine to be recognized as an independent medical specialty. Those involved in the intense process of obtaining formal recognition believe this will occur in the next year. PMID- 7486376 TI - Delayed presentation of uterine perforation. AB - A 30-year-old woman presented to the emergency department with nonspecific abdominal pain 17 days after voluntary vacuum aspiration interruption of a first trimester pregnancy. Physical examination and laboratory evaluation demonstrated mild diffuse abdominal tenderness without peritoneal signs and a mildly increased WBC count, respectively. When the patient's condition failed to improve she was admitted, and exploratory laparotomy was performed 5 days later. It revealed two uterine perforations, one of which contained incarcerated omentum with associated ischemic small bowel and complete small-bowel obstruction. Although uterine perforation with intraabdominal injury is a well-described complication of vacuum aspiration termination of pregnancy, most postabortion perforations go undetected. Delayed presentation of an acute complication is rarely observed. PMID- 7486377 TI - Acute accidental lindane ingestion in toddlers. AB - Lindane toxicity has been reported to occur mostly by way of dermal exposure. Cases of ingestion in which blood levels have been determined are rare. We present three such cases, along with a comparison of cases in the literature with respect to blood level half-lives and correlation with signs of toxicity. Emergency physicians can prevent acute ingestion by educating patients on the proper use of lindane and by selecting less toxic scabicidal agents. PMID- 7486378 TI - Successful heart transplantation from a victim of carbon monoxide poisoning. AB - Heart transplantation has become a highly successful, life-saving treatment for a number of otherwise fatal heart diseases. A major limiting factor in the growth of transplantation surgery has been the relative lack of suitable donor organs, and the appropriate criteria for selection of donor organs have been a topic of significant interest. Despite relatively favorable survival rates in the few patients who have received organs from victims of many types of poisonings and drug overdoses, patients dying of toxicologic causes are not usually considered suitable organ donors. Some centers routinely reject such individuals. Criteria for donor selection continue to be vague, unclear, or nonexistent in regard to organ transplantation from victims of all types of poisoning and toxic exposures. Carbon monoxide (CO) is a ubiquitous poison, and although victims of CO poisoning have occasionally served as suitable organ donors, heart transplantation in this scenario is still a very rare event. We describe the successful transplantation of the heart from a CO poisoning victim--to our knowledge, only the third such transplantation. Because the emergency department is a critical site for organ procurement, emergency physicians must be aware that patients dying of CO exposure may be acceptable organ donors. PMID- 7486379 TI - My friend Steve. PMID- 7486380 TI - Another positive-pressure tip. PMID- 7486381 TI - Ventricular fibrillation in pediatric cardiac arrest. PMID- 7486382 TI - Emergency medicine and health care reform. PMID- 7486383 TI - Relief of capsaicin contact dermatitis. PMID- 7486384 TI - No support for intravenous lidocaine airway reflex suppression during rapid sequence intubation. PMID- 7486385 TI - Kombucha "mushroom" hepatotoxicity. PMID- 7486386 TI - Pulse oximetry monitoring: blinded by the light, lulled by the beeps. PMID- 7486387 TI - Computed tomographic morphometry of the lumbosacral spine of dogs. AB - In a 5-year prospective study, computed tomographic (CT) morphometry of the lumbosacral vertebral canal was performed on 42 large-breed dogs (21 controls and 21 dogs with lumbosacral stenosis). Dogs were allotted to 4 groups. Group 1 (n = 13) consisted of cadaver specimens obtained from dogs that died or were euthanatized for reasons unrelated to the spine; group 2 (n = 8) consisted of live dogs with no history of clinical signs related to the spine and with normal neurologic examination findings; group 3 (n = 10) consisted of dogs with surgically confirmed lumbosacral stenosis; and group 4 (n = 11) consisted of dogs with suspected lumbosacral stenosis that were managed conservatively. The CT scans were performed, using 5-mm contiguous slices obtained perpendicular to the vertebral canal, from the midbody of the 5th lumbar vertebra through the caudal endplate of the sacrum (L5-S3). Lumbosacral lordosis was minimized in all dogs by positioning them in dorsal recumbency with the hind limbs flexed. A tuberculin syringe calibration phantom was placed within the scanning field of view, parallel to the axis of the spine. In each dog, 11 CT slice locations within the lumbosacral spine were evaluated. At each slice location, sagittal plane diameter, dorsal plane diameter, and transverse area of the vertebral canal, vertebral body, and calibration phantom were measured, using the CT computer's software programs for distance and area calculation. Window/level settings were constant, and all measurements were made by the same operator (JCJ). Accuracy of calibration phantom CT measurements was 100% for sagittal and dorsal plane diameter and was 85% for transverse area.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7486388 TI - Jejunal microvasculature of the llama and alpaca. AB - The vasculature of the jejunum was studied in 6 llamas and 1 alpaca, using a combination of microangiography, standard light microscopy, and vascular cast imaging. The casts were examined by use of scanning electron microscopy and low power dissecting microscopy. After administration of 40,000 IU of heparin, all animals were euthanatized by administration of an overdose of sodium pentobarbital. Three sections of jejunum and their respective arcuate vessels were isolated from each animal. One section was immediately placed in formalin for later H&E staining. The second and third sections were placed in warm saline solution, and the vasculature was flushed free of all blood by repeated infusions of the solution. Once flushed of all blood, one section was infused with a radio opaque medium and subsequently evaluated by microangiography, and the remaining section was perfused with a methylmethacrylate polymer for creation of vascular casts. The arcuate vessels branched into extensive primary and secondary arcades prior to giving rise to the marginal rete. Muscular arteries and small veins left the marginal rete and penetrated the tunica serosa and tunica muscularis to provide nutrients or drain the mesenteric angle, respectively, or entered into the circumferential submucosal network. The primary penetrating vessels in the submucosa formed an extensive submucosal plexus that supplied the tunica serosa, tunica muscularis, and tunica mucosa. The primary penetrating vessels anastomosed with vessels from oral and aboral sections and with their counterparts from the opposite side at the antimesenteric border.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7486389 TI - Heritability of the depth of the opening of the ciliary cleft in Samoyeds. AB - Ocular examination, including gonioscopy and ultrasonographic biometry (biological measurement), was performed in healthy, purebred Samoyeds. When the dogs were subclassified according to the degree of kinship with cases of primary angle-closure glaucoma, it was found that the opening of the ciliary cleft was wider in dogs less related to dogs with primary angle-closure glaucoma than in more closely related dogs. Furthermore, multivariate analysis of the material indicated that parentage of a dog has a substantial effect on the intraocular distances studied in this investigation (relative depth of the opening of the ciliary cleft, relative corneal thickness, relative anterior chamber depth, relative lens thickness, and relative length of the vitreous body). Finally, heritability of the relative depth of the opening of the ciliary cleft was estimated at 56% and that of the common environmental factor at 19%. The results are indicative of a hereditary component in primary angle-closure glaucoma in the Samoyed breed. PMID- 7486390 TI - Effects of various risk factors on plasma protein and serum immunoglobulin concentrations of calves at postpartum hours 10 and 24. AB - In an attempt to identify important predictors of failure of passive immunoglobulin transfer (< 800 mg of IgG/dl), identify calves with failure of passive immunoglobulin transfer, and determine the effects of a colostrum supplement, blood samples were collected from 263 calves at postpartum hours 10 and 24. Calves of dams diagnosed with mastitis had lower mean plasma protein and IgG concentrations at 10 (P < 0.05) and 24 (P < 0.01) hours. Plasma protein and IgG concentrations were similar for single and twin calves at 10 hours, but IgG concentration at 24 hours was higher (P < 0.01) in twin calves. Calves born to dams that had dystocia had numerically lower mean plasma protein and IgG concentrations than did calves born to dams that had normal delivery. However, observed differences were small and, after adjustment for other important factors, these differences were not significant. Age of dam was associated with plasma protein (P < 0.05) and IgG (P < 0.10) concentrations at 10 hours, but had no effect at 24 hours. Plasma protein and IgG concentrations decreased as calves were born later in the calving season, although the association of birth date with IgG concentration at 24 hours was marginal (P = 0.07). Calf sex, dam body condition score, and birth weight were not related to plasma protein or IgG values. The sensitivity and specificity of a cutoff value of 4.8 g of protein/dl of plasma, measured at 10 hours, for diagnosing failure of passive immunoglobulin transfer at 10 hours were 78 and 94%, and for diagnosing failure of passive immunoglobulin transfer at 24 hours were 88 and 73%, respectively. A colostrum supplement administered to calves with low plasma protein concentration at 10 hours had no effect on plasma protein or IgG values at 24 hours or on preweaning morbidity and mortality. PMID- 7486391 TI - Passive immune status at postpartum hour 24 and long-term health and performance of calves. AB - We quantified the effect of passive immune status on pre- and postweaning health and growth performance of calves raised in a beef production environment. Blood samples were collected at postpartum hour 24 from 263 crossbred calves for determination of plasma protein (PP) and serum IgG concentrations. Serum IgG concentration was classified as adequate (> 1,600 mg/dl), marginal (800 to 1,600 mg/dl), or inadequate (< 800 mg/dl). Plasma protein concentration was classified as adequate (> or = 4.8 g/dl) or inadequate (< 4.8 g/dl). Morbidity and mortality events in the study population were monitored from birth to weaning, and after weaning throughout the feeding period. The lowest concentrations of serum IgG and PP were observed among calves that experienced morbidity or mortality prior to weaning. Calves that experienced morbidity in the feedlot had lower 24-hour PP values, but had IgG concentration similar to that in calves that were not observed to be ill during the feeding period. Calves classified as having inadequate IgG concentration were at greater risk of preweaning mortality (odds ratio [OR] = 5.4), neonatal morbidity (OR = 6.4), and preweaning morbidity (OR = 3.2), compared with calves classified as having adequate IgG concentration at 24 hours. Calves classified as having inadequate PP concentration at 24 hours had a greater risk of morbidity (OR = 3.0) and respiratory tract morbidity (OR = 3.1) while in the feedlot, compared with calves classified as having adequate PP concentration. The effects of 24-hour passive immune status on calf growth were indirect through effects on morbidity outcomes.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7486393 TI - Serologic studies of experimentally induced Salmonella choleraesuis var kunzendorf infection in pigs. AB - Two indirect ELISA containing outer membrane protein (OMP) and lipopolysaccharide (LPS) antigens from a field isolate of Salmonella choleraesuis var kunzendorf were developed and evaluated in experimentally infected and uninfected control pigs. Experimentally induced infection with S choleraesuis was successfully established in 10 pigs by oral inoculation with 10(8) organisms, and 3 pigs died of clinical salmonellosis at postinoculation (PI) weeks 1, 2, and 4. Swab specimens from tonsils, nostrils, and rectum of pigs were obtained for culture, and sera were evaluated at weekly intervals for 9 weeks after inoculation. The ELISA containing OMP and LPS antigens with either anti-swine IgG or protein albumin-to-globulin ratio (antiglobulin) conjugates were standardized for serologic evaluation. All 4 ELISA (2 OMP and 2 LPS) detected seroconversion by PI week 3 and had sensitivities and specificities of 97.8 and 88.8, 100 and 100, 95.6 and 88.8, and 93.3 and 72.5%, at their ideal cutoff points (negative mean optical density +2 SD). There was excellent agreement between all 4 ELISA systems as determined by kappa values. Cultures of fecal, tonsil, and nasal swab specimens were positive for S choleraesuis until the fourth week of infection. Fecal swab specimens from 1 pig were positive for S choleraesuis until PI week 7. Persistent infection after antemortem culture results were negative was detected by all 4 ELISA, which indicated consistently high titers until the end of PI week 9.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7486392 TI - Field trial to evaluate immunogenicity of a glycoprotein I (gE)-deleted pseudorabies virus vaccine after its administration in the presence of maternal antibodies. AB - A field trial was conducted on a commercial swine farm quarantined because of infection with pseudorabies virus. The purpose was to investigate, in growing pigs born to hyperimmunized sows, the immunogenicity of a vaccine with a glycoprotein I (gE) deletion. One hundred twenty pigs were assigned at random to 1 of 3 vaccination schedules at ages: 8 and 12 weeks; 8, 12, and 14 weeks; and 8, 12, and 16 weeks. Immune response was measured at 8, 12, 14, 16, and 18 weeks, using the serum neutralization test, a screening ELISA, and assays of IgG and IgA in serum and nasal secretions. Results of the serum neutralization test and the screening ELISA indicated that, for pigs vaccinated only at 8 and 12 weeks, the percentage of pigs with pseudorabies virus serum antibodies decreased substantially by 18 weeks; for pigs given a booster at 14 or 16 weeks, the prevalence of serum antibodies at 18 weeks was higher, with 16-week booster vaccination eliciting the best response. At each age, nasal IgA and IgG values were highly correlated (r > or = 0.70), as were serum IgA and IgG values; correlations of serum with nasal IgA and IgG values were somewhat lower (approx range, r = 0.40 to 0.70). Nevertheless, an increase in serum IgA or IgG values on vaccination was no guarantee of an increase in nasal IgA or IgG values. For serum and nasal mucosal antibodies, a poor immune response was associated with high quantities of maternally derived antibodies. Vaccination at 16 weeks was necessary to ensure eliciting of an immune response in almost all pigs. PMID- 7486394 TI - Persistent anthelmintic activity of ivermectin against gastrointestinal nematodes of cattle. AB - Two experiments were conducted to determine persistence of ivermectin (IVM) anthelmintic activity. In the first experiment, the injectable (INJ) formulation at a dosage of 200 micrograms/kg of body weight and pour-on (PO) formulation at a dosage of 500 micrograms/kg were compared in 5 groups of calves (n = 6/group). Calves were treated at 14 and 7 days prior to experimentally induced infection. Experimental groups were: 1--untreated, infected controls; 2--IVM PO, day -14; 3- IVM PO, day -7; 4--IVM INJ, day -14; and 5--IVM INJ, day -7. Calves were necropsied on days 28 and 29 after infection. Results of this experiment indicated a high degree of efficacy of IVM PO product (93.8%) for all nematodes up to 14 days, in contrast to poor activity for IVM INJ (26.3%) for all nematodes. Seven-day persistence was excellent for both IVM formulations against Ostertagia ostertagi and Cooperia spp, but a lower degree of efficacy (77.0% for PO and 88.5% for INJ) was observed against Haemonchus placei. In the second experiment, persistent efficacy of IVM PO in preventing establishment of O ostertagi inhibited larvae was observed. In 3 trials, groups of 4 treated and 4 untreated control calves were exposed to natural pasture infection at 0 to 7, 10 to 17, and 20 to 27 days after treatment. Calves were necropsied 15 to 16 days after removal from pasture. After the day 0 to day 7 infection exposure, IVM PO efficacy was 100% for all stages of O ostertagi, 3 adult male Cooperia spp, and Oesophagostomum radiatum; 94.9% for H placei; and 98.2% for Cooperia spp adult females.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7486397 TI - Comparison of estrogen and progesterone receptor expression in normal and tumor mammary tissues from dogs. AB - Concentrations of estrogen (ER) and progesterone (PR) receptors were measured by radioreceptor assay in tumor (n = 319) and normal (n = 166) mammary tissue from 248 bitches. Correlations between ER and PR and between receptor expression in tumor and normal mammary tissue from the same bitches were evaluated. The influence of tumor, clinical, or hormonal variables on receptor expression also was studied. Approximately 80% of tumor and 95% of normal mammary tissue expressed detectable concentrations of ER, PR, or both. Direct correlation was found between ER and PR concentrations in normal and tumor tissues. Median ER concentrations were significantly higher (46 +/- 47 fmol/mg of cytosolic protein vs 27 +/- 24 fmol/mg of cytosolic protein; P = 0.0002) in normal than in tumor tissue. On the other hand, PR concentrations were significantly higher (57 +/- 52 fmol/mg vs 77 +/- 99 fmol/mg; P = 0.03) in tumors (especially benign tumors) than in normal tissue. Poorly differentiated malignant tumors expressed lower concentrations of receptors than did benign or well differentiated malignant tumors. The ER and PR concentrations decreased with increasing size of the lesion. Hormonal status of the bitch significantly (P < 0.05) influenced receptor expression in normal tissue: bitches in the luteal phase of the estrous cycle had higher concentrations of ER (69 +/- 62 fmol/mg) than did ovariectomized bitches (24 +/- 19 fmol/mg) or bitches in anestrus (38 +/- 45 fmol/mg) or the follicular phase (13 +/- 7 fmol/mg).(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7486395 TI - Abortions, fetal death, and stillbirths in pregnant pygmy goats inoculated with tachyzoites of Neospora caninum. AB - Neospora caninum-induced abortion is a major production problem in the dairy cattle industry in the United States and worldwide. Abortions attributable to naturally acquired N caninum infection also have been observed in pygmy goats. We studied experimentally induced infections with N caninum in pregnant pygmy does to determine whether abortions attributable to N caninum infection would occur after inoculation. Seven pregnant pygmy does (1 control doe and 6 inoculated with N caninum) were studied. The control doe remained clinically normal throughout the study and delivered 2 healthy kids. Abortion, fetal death, and stillbirths were observed in some pregnant does inoculated with N caninum. Two pregnant pygmy does inoculated with N caninum early in gestation (day 51) had fetuses that died and were aborted, or died and were reabsorbed. Neospora caninum tachyzoites and lesions were observed in the brain, spinal cord, and heart of aborted fetuses; parasites also were isolated from the placenta. Four additional pregnant pygmy does (2 inoculated at mid-gestation [day 85], and 2 at late gestation [day 127]) did not abort after inoculation. However, 1 doe inoculated during mid-gestation delivered a stillborn fetus that had died about 1 week prior to parturition. This kid was congenitally infected with N caninum. Neospora caninum was isolated from the placentas of all inoculated does examined. Neonatal neosporosis was not observed in live-born kids, nor were stages of N caninum isolated from any live born kid. Does did not undergo abortion or have congenitally infected kids when they were rebred and evaluated for neosporosis. PMID- 7486396 TI - Comparison of methods for estimation of Toxoplasma gondii-specific antibody production in the aqueous humor of cats. AB - Intraocular production of Toxoplasma gondii-specific antibody in cats has been estimated by comparing the ratio of T gondii-specific antibody in aqueous humor and serum with the ratio of total immunoglobulins in serum and aqueous humor (Goldmann-Witmer coefficient; aqueous antibody coefficient; C value). It has been proposed that in human beings, comparison of the ratio of T gondii-specific antibody in aqueous humor and serum with the ratio of antibodies against a nonocular pathogen in serum and aqueous humor is more accurate than methods using total immunoglobulin quantification. We developed an ELISA for detection of calicivirus-specific antibodies in the serum and aqueous humor of cats. By evaluating calicivirus-specific antibody concentrations in the aqueous humor of healthy and diseased cats, calicivirus was assessed as a nonintraocular pathogen. The ratio of T gondii-specific antibodies in the aqueous humor and serum and the ratio of calicivirus-specific antibodies in serum and aqueous humor were evaluated as a means of estimating intraocular T gondii-specific antibody production. A field strain of feline calicivirus was isolated, cultured, and purified. A calicivirus-specific IgG ELISA was developed for detection of feline calicivirus-specific IgG in serum and aqueous humor. Calicivirus-specific IgG was measured in the serum and aqueous humor from 3 groups of control cats. Results suggested that calicivirus is a nonintraocular pathogen in cats and that calicivirus IgG detected in aqueous humor is attributable to leakage across a damaged blood-ocular barrier.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7486398 TI - Study of transneuronal passage of pseudorabies virus in rat central nervous system by use of immunohistochemistry and in situ hybridization. AB - We studied the uptake and sequential transneuronal passage of pseudorabies virus (PRV) in rat CNS by use of a combination of immunohistochemistry and in situ hybridization. Protocols for rapid detection of PRV by immunohistochemistry and in situ hybridization in rats with PRV infection of the CNS after intranasal instillation of a wild-type strain of PRV were optimized in vitro, using porcine kidney-15 cells. Pseudorabies virus-specific hybridization signals appeared in the cytoplasm and nucleus of PRV-infected porcine kidney-15 cells by postinoculation (PI) hour 6. In tissue sections of PRV-infected rats, PRV nucleic acids were detected in areas of the rat brain in close proximity to the areas in which PRV antigens were evident. The PRV was initially found in the nucleus of trigeminal ganglion neurons at PI hour 24. At PI hour 72, PRV antigens were observed in the mid-brain, and 24 hours later, in the telencephalon. We also found evidence of specific progressive transsynaptic transmission of the virus, and, on the basis of that, we have constructed a map of the synaptic contacts and pathways in the brain. Therefore, combined use of immunohistochemistry and in situ hybridization was useful for characterizing the pathogenesis of PRV in the CNS of rats after intranasal inoculation, following a pattern that mimics PRV infection of the natural host. PMID- 7486399 TI - Effect of glucocorticoids on serum osteocalcin concentration in horses. AB - The effects of dexamethasone (0.2 mg/kg of body weight; IV, IM, and PO) and methylprednisolone acetate (120 mg given intra-articularly) on serum osteocalcin and cortisol concentrations were studied in 6 horses. Serum osteocalcin and cortisol concentrations were serially monitored after each treatment. A significant (P < 0.05) decrease in serum osteocalcin and cortisol concentrations was observed from 12 to 24 and 2 to 48 hours, respectively, after IV and IM administrations of dexamethasone. Serum osteocalcin and cortisol concentrations were significantly decreased from 6 to 48 and 3 to 72 hours, respectively, after oral administration. In contrast, a change in serum osteocalcin concentration was not detected after intra-articular administration of methylprednisolone. Oral, IV, or IM treatment with 0.2 mg of dexamethasone/kg caused a decrease in serum osteocalcin concentration in horses. PMID- 7486400 TI - Pharmacologic interaction of furosemide and phenylbutazone in horses. AB - The effect of premedication with phenylbutazone on systemic hemodynamic and diuretic effects of furosemide was examined in 6 healthy, conscious, mares. Mares were instrumented for measurement of systemic hemodynamics, including cardiac output and pulmonary arterial, systemic arterial, and intracardiac pressures, and urine flow. Each of 3 treatments was administered in a randomized, blinded study; furosemide (1 mg/kg of body weight, IV) only, phenylbutazone (8.8 mg/kg, PO, at 24 hours and 4.4 mg/kg, IV, 30 minutes before furosemide) and furosemide, or 0.9% NaCl. Phenylbutazone administration significantly attenuated, but did not abolish, the diuretic effect of furosemide. Phenylbutazone completely inhibited the immediate effect of furosemide on cardiac output, stroke volume, total peripheral resistance, and right ventricular peak pressure. Premedication with phenylbutazone did not inhibit equally the diuretic and hemodynamic effects of furosemide, indicating that some of furosemide's hemodynamic effects are mediated by an extrarenal activity of furosemide. PMID- 7486401 TI - Effect of phenylbutazone and flunixin meglumine on acute toxic mastitis in dairy cows. AB - A double-blinded randomized prospective clinical trial was designed to evaluate the effectiveness of flunixin meglumine and phenylbutazone for treatment of acute toxic mastitis in dairy cows. All cows were treated 4 times at 12-hour intervals by intramammary infusion of gentamicin (150 mg). A total of 45 dairy cows with toxic mastitis were randomly assigned to 1 of 3 treatment groups: group 1 (control), saline solution, IV; group 2, 1 g of flunixin meglumine, IV; or group 3, 4 g of phenylbutazone, IV. Physical examination and udder variables were assessed at initial examination and 24 hours later. Milk production was recorded at regular intervals from 1 week before until 10 weeks after development of mastitis. Rear quarters (34/45) were more commonly affected than front quarters. Thirty-five cows returned to the herd, 9 cows were culled, and 1 cow died. There were no significant differences among treatment groups in the need for further treatment or outcome. Klebsiella spp (18/45) and Escherichia coli (16/45) were the most common pathogens isolated by culture of milk from affected quarters. The overall bacteriologic cure rate on days 7 and 14 was 64 and 75%, respectively. At the time of initial examination, cows of the control group had higher rectal temperature than did cows of the flunixin group. At the examination 24 hours later, the rectal temperature of cows in all treatment groups was lower than the temperature at initial examination; at that time (24 hours), however, there were no significant differences in temperature among the treatment groups.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7486402 TI - Pharmacokinetics of enrofloxacin in clinically normal dogs and mice and drug pharmacodynamics in neutropenic mice with Escherichia coli and staphylococcal infections. AB - Pharmacodynamic variables of enrofloxacin were investigated in a neutropenic mouse Escherichia coli and staphylococcal thigh infection model. Enrofloxacin pharmacokinetics in clinically normal mice and dogs were compared to confirm that doses evaluated in the mouse model would include enrofloxacin doses appropriate for use in dogs. Mice were made neutropenic by treatment with cyclophosphamide and injected in the thigh muscle with approximately 10(6) colony-forming units of E coli (n = 2) or a staphylococcal (n = 2) clinical isolate. Enrofloxacin dosages tested ranged from 0.78 to 50 mg/kg of body weight and 6.25 to 200 mg/kg in the E coli and staphylococcal infection trials, respectively. In each 24-hour dosage trial, enrofloxacin was administered SC as a single dose or in divided doses given every 3, 6, or 12 hours. Comparison of log10 colony-forming units per thigh muscle in untreated control mice and mice treated with enrofloxacin was used as a measure of efficacy. Two-way ANOVA was used to determine that the enrofloxacin total dose, but not the dose frequency, was significant in determining drug efficacy. Pharmacokinetic values analyzed by use of multivariant stepwise linear regression analysis indicated that the area under the concentration-time curve, but not time above minimum inhibitory concentration, was significant in predicting efficacy of enrofloxacin treatment. We conclude that enrofloxacin killing of E coli and staphylococci is concentration dependent and not time dependent. PMID- 7486403 TI - Effects of short-term restraint stress on plasma concentrations of catecholamines, beta-endorphin, and cortisol in gilts. AB - The effects of short-term restraint stress, by means of snaring, on plasma concentrations of catecholamines, beta-endorphin, and cortisol were studied in 6 gilts. A catheter was inserted into the jugular vein, and 2 blood samples were collected before onset of stress. Thereafter, a hog snare was applied, and blood samples were collected at 0.5, 2, and 3.5 minutes after the start of snaring. Plasma epinephrine and norepinephrine concentrations increased (P < 0.001) within 0.5 minute after start of restraint and decreased thereafter. Plasma concentration of beta-endorphin increased (P < 0.05) within 2 minutes after start of restraint, whereas that of cortisol increased (P < 0.05) 3.5 minutes after start of restraint. Taken together, short-term stress, such as snaring may increase the plasma concentration of catecholamines, beta-endorphin, and cortisol in pigs. PMID- 7486404 TI - Effects of short-term cortisol infusion on growth hormone-releasing hormone stimulation of growth hormone release in sheep. AB - Excess production or long-term administration of glucocorticoids is detrimental to longitudinal growth in people and rats. A portion of this effect is attributed to cortisol inhibition of growth hormone (GH). Glucocorticoid effects are usually studied in subjects under long-term treatment with synthetic, more potent glucocorticoids, and, to the authors' knowledge, have not been examined in domestic animals. We sought to examine the effects of cortisol infusion on GH release in sheep. Cortisol infusion into castrated, male Suffolk sheep (1 to 1.5 years old) caused a significant (P < 0.0001) increase in cortisol concentration. Basal GH release was not affected over the 4-hour period of infusion. Growth hormone-releasing hormone administration stimulated GH release in both groups (P < 0.001); however, the control group had a greater response to growth hormone releasing hormone than did the cortisol infused group (P < 0.0001). These results were duplicated in cultured sheep pituitary cells. Cortisol inhibition of GH release may be mediated via enhanced somatostatin release, owing to a direct inhibition of somatotrope function, or a combination of both mechanisms. Because of effects of stress and disease in increasing cortisol concentration, additional study of the mechanisms for cortisol inhibition of GH release in sheep needs to be performed. PMID- 7486405 TI - Cardiovascular, hormonal, and metabolic responses to severe prolonged hemorrhage in adult sheep. AB - Over a 54-hour period, blood was removed from 8 adult sheep (body weight, 38.1 +/ 0.5 kg, mean +/- SEM) in 9 episodes, 5 on day 1, 3 on day 2, and 1 on day 3. Cumulative blood loss was 1,630 +/- 63, 2,380 +/- 71, and 2,693 +/- 69 ml on days 1, 2, and 3, respectively. Blood samples (20 ml) were collected from 5 control ewes (33.8 +/- 2.8 kg) at equivalent times. Over the first day, mean arterial blood pressure decreased in the hemorrhaged sheep from 101 +/- 2 mm of Hg to 76 +/- 5 mm of Hg, but returned to control values by the beginning of the second day and, thereafter, was not different from control values. Heart rate was increased after the first hemorrhage episode and remained high throughout the entire protocol. Over the entire period, there were statistically significant decreases in hematocrit, plasma osmolality, sodium, total calcium (P < 0.001), potassium, and chloride values (P < 0.05). There was no change in plasma phosphate, bicarbonate, creatinine, or magnesium concentrations and an increase in plasma urea nitrogen (P < 0.001) concentrations. Plasma arginine vasopressin concentration was increased significantly (P < 0.001) over the entire period. Plasma ACTH concentration was significantly (P < 0.05) increased over time, but only some values on day 1 were significantly outside the normal range of the control group data. Because of wide variation between sheep, the group data for aldosterone were not significantly different from control values.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7486407 TI - Histologic evaluation of tissue reactivity and absorption in response to a new synthetic fluorescent pigmented polypropylene suture material in rats. AB - The degree and type of tissue reactivity and the absorption of a new suture material was determined by implantation within rat gluteal muscles. Amount and type of tissue inflammatory reaction was compared among the new suture material, polypropylene, and coated polyamide. Histologic evaluation of the tissues in which sutures were implanted indicated that the new suture material, polypropylene, and coated polyamide had similar amounts and types of reaction at 30 days or less after implantation, but differed after 30 days. The new suture material and polypropylene had an inflammatory reaction zone measuring less than 25% of the high-power field after 60 days, but the coated polyamide still induced reaction greater than 45% of the field at 90 days. At 60 and 90 days after implantation, the new suture material and polypropylene induced a mature fibrous reaction; the reaction to coated polyamide was either immature fibrous or granulomatous, depending on whether there was rupture of the suture coat. There was no observable absorption of the new suture material at 90 days. This study indicated that the new suture material is nonabsorbable and is minimally reactive in rat muscle. The tissue reactions induced by this suture material are similar to those of polypropylene and significantly less than those induced by coated polyamide after 30 days following implantation. PMID- 7486406 TI - Effects of stromelysin activity on proteoglycan degradation of canine articular cartilage explants. AB - We investigated whether stromelysin activity in the medium of canine articular cartilage explants is associated with proteoglycan degradation in these explants. Cartilage explants were treated with recombinant human interleukin 1 alpha (rh-IL 1 alpha), lipopolysaccharide, or canine monocyte-conditioned medium. Proteoglycan synthesis and degradation were measured. Metalloproteinase activity (inhibitable by tissue inhibitor of metalloproteinase 2) in the culture medium was measured by use of fluorimetry with a quenched fluorescent substrate. Western blots of the medium were probed with polyclonal antibodies to human stromelysin, collagenase, and gelatinase. Neither metalloproteinase activity nor proteoglycan degradation were inducible in canine cartilage explants treated with rh-IL-1 alpha. However, proteoglycan synthesis was significantly (P < 0.05) decreased by concentrations of 10 and 100 ng of rh-IL-1 alpha/ml. Metalloproteinase activity in the medium accompanied proteoglycan degradation of cartilage treated with lipopolysaccharide and monocyte-conditioned medium. The metalloproteinase released into the medium was identified as prostromelysin by results of western blotting. PMID- 7486409 TI - Agrees with and adds to report of a link between equine herpesvirus 2 and chronic pulmonary disease in horses. PMID- 7486408 TI - Effects of feeding fumonisin B1 in lactating sows and their suckling pigs. AB - The mycotoxin fumonisin B1 (FB1) produced by Fusarium moniliforme in corn causes pulmonary edema in finishing swine. Effects of lower nonlethal amounts and effects in lactating sows with suckling pigs are unknown. An initial study was conducted to determine a nonlethal concentration of FB1 for lactating sows; whether ingested FB1 could be detected in the milk; and whether toxicosis could be detected in the pigs, as determined by necropsy. Another study was conducted to determine toxicosis in the pigs by measuring liver sphinganine-to-sphingosine ratio, and whether ingested FB1 affected T-lymphocyte function in sows and their pigs. Furthermore, sows of this study were maintained in controlled hot (27 to 32 C, 50 to 70% relative humidity) and thermoneutral (21 C, 55% relative humidity) environments to determine whether high temperature exacerbated the effects of FB1. In the first study, 100 micrograms of FB1/g of corn soybean meal diet was found to be nonlethal when fed for 14 days. Fumonisin B1 was not detected in the milk at 30 ppb and lesions were not found in the necropsied pigs, including 1 from a sow that died of porcine pulmonary edema syndrome after ingesting FB1 at a concentration of 175 ppm. In the second study, differences in liver sphinganine to-sphingosine ratio of pigs were not found. Expressions of cell surface antigens on blood lymphocytes and lymphocyte proliferation response to various mitogens were not affected by FB1 or high temperature in sows or their pigs.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7486410 TI - Basic behavioral science research for mental health. A national investment. Emotion and motivation. National Advisory Mental Health Council. PMID- 7486411 TI - Increased incidence of nosocomial infections in obese surgical patients. AB - Obesity has long been considered a potential risk factor for poor outcome following surgical procedures. However, controversy exists regarding the clinical impact of this problem because of a paucity of data regarding the incidence and risk of nosocomial infections in obese surgical patients. This retrospective study was undertaken to compare the nosocomial infection rate in obese and normal weight surgical patients. All patients undergoing general, urologic, vascular, thoracic, or gynecologic surgical procedures between October 1 and December 31, 1991, were reviewed. Nosocomial infection data were obtained from the Department of Hospital Epidemiology. A total of 849 patients were evaluated, of which 536 (63%) were normal weight (BMI < 27 kg/m2), 175 (21%) were obese (BMI 27-31 kg/m2), and 138 (16%) were severely obese (BMI > 31 kg/m2). Age, mortality, and American Society of Anesthesia (ASA) risk scores did not differ among the three groups. There were significant increases in the number and percent of nosocomial infections in the obese populations, with rates of 0.05 per cent in normal weight, compared to 2.8 per cent and 4.0 per cent in obese and severely obese groups (P < 0.01). Infections consisted of seven wound infections, five C. difficile infections, one pneumonia, and three bacteremias. No differences in distribution between groups were evident. Mortality was similar among the groups. These data support the hypothesis that obesity is a significant risk factor for clinically relevant nosocomial infections in surgical patients. PMID- 7486412 TI - Syncope as etiology of road crashes involving elderly drivers. AB - A significant number of elderly drivers present without discernible external cause for their road crash. We hypothesize that syncope may contribute to their crash etiology and prospectively evaluated drivers older than 60 years with unexplained crash, presenting to our trauma center during a 1-year period. Prehospital data and prior medical history were obtained on all, followed by a standard syncope work-up. Of 79 drivers ages 60-98, 58 (73%) were at fault in the crash. In 33 (57%), the crash etiology could not be determined. Thirteen had prior syncope history. Workup was negative in two patients, and six died or were transferred before evaluation. Twenty-five of 33 (76%) had positive workups for syncope: cardiac causes in 10; neurologic in seven; psychiatric in five, endocrine in two; one indeterminant. Ten of 18 who survived had license revocation for medical reasons. Syncopal episodes are a frequent contributor to unexplained crashes among elderly drivers and should be suspected whenever external causes are not apparent. PMID- 7486413 TI - The effect of epidural analgesia on the return of peristalsis and the length of stay after elective colonic surgery. AB - Epidural analgesia has been shown to improve pulmonary function and analgesia after abdominal surgery. Although epidural analgesia may increase colonic motility, its effect on the clinical outcome following colonic surgery is unclear. Therefore, the purpose of this study was to determine the effect of epidural analgesia on return of peristalsis and length of stay after elective colonic surgery as compared with traditional analgesia. A total of 102 patients undergoing elective colonic surgery (66 left-sided, 36 right) between September 1991 and April 1993 were retrospectively studied. Variables recorded were location of colon anastomosis, type of analgesia, time until return of colonic peristalsis, length of epidural administration, and length of hospital stay. Forty-one patients received epidural and 61 received traditional analgesia. Data was analyzed with a Student's T test (P set < 0.05) and Spearman's correlation coefficient where appropriate. No statistically significant difference was found between epidural and traditional analgesia with regard to return of peristalsis or length of stay. Similarly, when considering the location of the anastomosis (left versus right colon), no statistically significant difference was demonstrated between the two groups. Also, there was no increased incidence in anastomotic leaks or mortality. PMID- 7486414 TI - Method of salvaging long-term dialysis catheters. AB - Hemodialysis is an important component of chronic renal replacement therapy, which is increasingly being provided with indwelling venous catheters. Catheter malfunction is commonly dealt with using Urokinase instillation or endovascular catheter stripping. We describe the application of a simple technique that allows the indwelling dialysis catheter to be replaced in a subcutaneous tunnel following manipulation for flow problems. Function was restored in all catheters without occurrence of tunnel infection or catheter-related sepsis. Preliminary results offer evidence of the efficacy of the technique in salvaging dialysis catheters, especially in patients with difficult vascular access. PMID- 7486415 TI - Pelvic congestion syndrome: a new approach to an unusual problem. AB - Pelvic congestion syndrome (PCS) is an uncommon and frequently overlooked cause of debilitating pelvic pain. The well-described clinical presentation is that of pain and fullness exacerbated by prolonged standing, coitus, and in the premenstrual period in multiparus women. Physical signs include vulvar varices that can communicate with the saphenous vein in the groin, causing thigh or buttock varices. The diagnosis is usually confirmed by ovarian vein venography demonstrating reflux to the ovaries and often into the thigh with erect positioning and valsalva. Standard surgical treatment is bilateral ovarian vein ligation and excision or ligation of as many collaterals as possible. The traditional approach is bilateral retroperitoneal incisions, with medial rotation of the viscera. We report the first patient managed transperitoneally using minimally invasive techniques. Our case suggests that this approach can easily and safely be performed by surgeons experienced in laparoscopic surgery with the advantages of improved cosmesis, less postoperative pain, and rapid convalescence typical of other minimally invasive procedures. Additionally, it provides the opportunity to perform diagnostic laparoscopy as well. PMID- 7486416 TI - Giant cavernous hepatic hemangiomas in adults: enucleation under selective blood inflow control. AB - Among hepatic hemangiomas (the most common solid hepatic tumor in adults), "giant hemangiomas" are those defined by a diameter greater than 4 cm. The natural history of such lesions is unclear, and many surgical procedures have been described. In five women and one man (mean age 49), we successfully used enucleation of the tumor under blood inflow control. This technique was made possible by a cleavage plane between the hepatic parenchyma and the hemangioma. This straightforward and safe procedure required a low rate of transfusion (mean 200 mL/patient) and spared a maximum amount of hepatic tissue. PMID- 7486417 TI - Internal supravesical hernia: report of a case. AB - A patient with clinical signs of intestinal obstruction was found at celiotomy to have a loop of small intestine caught in the supravesical space between pubis and urinary bladder. This report aims to bring attention to vesical hernias, a very rare entity, as a possible cause of small bowel obstruction. PMID- 7486418 TI - Wolfgang Amadeus Mozart 1756-1791: a mysterious death. PMID- 7486419 TI - Pheocromocytoma and gastrointestinal bleeding. AB - A 46-year-old patient who presented with an abdominal mass and gastrointestinal bleeding associated with an extra-adrenal pheochromocytoma is discussed. He had no history of hypertension. An ill-defined mass was felt at the upper abdomen. CT and arteriography confirmed the presence of a large mass with calcifications in the right paravertebral region. Upper endoscopy revealed multiple varices at the third portion of the duodenum. Abdominal exploration revealed a huge tumor at the root of the small bowel mesentery with multiple arterial and venous vessels entering the third portions of the duodenum. The tumor was unresectable. Biopsy demonstrated a pheochromocytoma. The patient developed pulmonary edema in the immediate postoperative course. He recovered and was discharged home on Dibenzyline. He has been readmitted on various occasions with gastrointestinal bleeding and congestive heart failure. Presently he is working and feels relatively well 5 years after the operation. A review of the literature for gastrointestinal complications of pheochromocytomas was done. There is a scarcity of reports of gastrointestinal bleeding associated with pheochromocytomas. PMID- 7486420 TI - Rupture of splenic artery pseudoaneurysm. AB - Pseudoaneurysms of the splenic artery have been well described in association with pancreatic pseudocysts secondary to pancreatitis. We present a case of a ruptured splenic artery pseudoaneurysm 14 years after splenectomy for trauma which, at that time, was complicated by a subphrenic abscess. PMID- 7486421 TI - High cervical intravagal hypercellular parathyroid gland as the etiology of severe persistent primary hyperparathyroidism. AB - The preoperative localization and operative management of a high cervical intravagal parathyroid adenoma in a young woman with severe persistent primary hyperparathyroidism is presented. The etiology of persistent hyperparathyroidism is most frequently a missed adenoma that is often in an ectopic location; of these, undescended adenomas represent a small but important proportion. If clinical suspicion and preoperative localization procedures suggest an undescended gland, a direct surgical approach using a high cervical incision and a unilateral exploration is often successful. PMID- 7486422 TI - Herniography following indirect hernioplasty using the Marlex "mesh plug" technique. AB - In recent years, some surgeons have been performing indirect inguinal herniorrhaphies using a "mesh plug" technique without ligation or resection of the hernia sac. The authors have had an interest in herniography and questioned whether herniorrhaphy without ligation or resection of the indirect sac might result in a false positive herniogram or an abnormal filling defect within the peritoneal cavity. These questions were answered by the performance of herniography in five patients who had indirect hernias repaired by this method. No false positive herniograms were discovered, and the inverted sacs could not be identified within the peritoneal cavity. Therefore, herniography would still be a useful diagnostic tool in evaluating patients who have persistent or recurrent symptoms following operations wherein the hernia sac was left intact. PMID- 7486423 TI - Early complications and long-term survival in severely obese coronary bypass patients. AB - Our objective was to describe the short-term morbidity of coronary artery bypass operations and the effect of surgery plus aggressive cardiac rehabilitation on the long-term prognosis of severely obese patients with coronary artery disease. We investigated an inception cohort of 28 consecutive severely obese patients with three-vessel coronary disease followed on average for 51 months. The patients' age, preoperative and postoperative weight, risk factors, and cholesterol were measured. We performed coronary artery bypass surgery, then began aggressive cardiac rehabilitation programs. We recorded intraoperative data, perioperative deaths, complications, readmissions, and lengths of stay. Also, New York Heart classifications and use of anti-anginal or cholesterol lowering medications were noted. All patients were followed up. Despite high morbidity, long-term function and survival of severely obese CABG patients compares favorably with that of average patients. However, aggressive behavior modification fails to alter their postoperative weight or risk profile, placing them at risk for both second CABG procedures and continued obesity-related disease occurrences. PMID- 7486424 TI - Massive primary lipoma of the scrotum. AB - Most lipomas occurring in the scrotum originate and develop in the spermatic cord. In rare cases, however, lipomas originate in the fat cells of the subcutaneous tissues of the scrotal walls themselves. These primary lipomas of the scrotum vary in size. We describe a patient with a scrotal mass that, to our knowledge, is the largest scrotal lipoma originating in the tunica dartos reported in the English literature. PMID- 7486425 TI - Incidence and risk factors for early small bowel obstruction after celiotomy for penetrating abdominal trauma. AB - The reported overall incidence of postoperative Small Bowel Obstruction (SBO) is 0.69 per cent. This study examined the incidence and risk factors for early postoperative SBO for penetrating abdominal trauma (PAT), with early SBO defined as SBO within 6 months of operation. This was a prospective cohort study of patients admitted to a Level 1 Trauma Center from 5/91 to 12/93 who required celiotomy for PAT. Patients were followed at least 6 months for readmission for SBO to be considered evaluable. Of 341 patients undergoing celiotomy for PAT and surviving to discharge, 298 (87.4%) were evaluable. The overall incidence of early SBO was 7.4 per cent and varied between 2.3 per cent (nontherapeutic celiotomy) and 10.8 per cent (small/large bowel injury). All patients with celiotomy for PAT are at increased risk for early SBO compared with elective surgery patients. Those with small/large bowel penetration or gunshot wounds are at the highest risk. Previous abdominal surgery is not a risk factor for early SBO in PAT patients. Surgeons and Managed Care case managers should devote special attention to close follow-up in PAT patients, particularly those with the risk factors identified in this study. PMID- 7486426 TI - Hydrocele of the canal of Nuck: ultrasound appearance. AB - Swelling in the inguinal region of a woman may result from a number of conditions, including inguinal hernia, tumor (lipoma, leiomyoma, sarcoma), cyst, abscess, adenopathy, or hydrocele of the canal of Nuck. It may be difficult to make a specific diagnosis based upon the history and physical examination alone. This report describes a symptomatic woman in whom ultrasound was helpful in diagnosing a hydrocele of the canal of Nuck, which is the female counterpart of a spermatic cord hydrocele in the male. The ultrasound findings are described. PMID- 7486427 TI - Abdominal wall endometrioma in a laparoscopic trocar tract: a case report. AB - Extrapelvic endometriosis often presents as an atypical, painful abdominal mass often referred to the surgeon for diagnosis and treatment. The majority of extrapelvic endometriosis is found in surgical scars. We present a patient with an abdominal wall endometrioma that occurred in a laparoscopy trocar tract. To our knowledge, this is the first such case reported in the surgery or gynecology literature. With the increasing use of laparoscopy for gynecologic and general surgery procedures, this problem will likely become more common. The general surgeon should know how to diagnose and treat endometriomas arising in laparoscopic trocar tracts. PMID- 7486428 TI - Acute cholecystitis in diabetic patients. AB - Postoperative outcome and severity of acute cholecystitis in 32 diabetic patients (DM) who underwent urgent cholecystectomy were compared on a case-control basis with 32 nondiabetic age/sex matched controls. There was no difference in incidence of renal and lung diseases or duration of acute symptoms before surgery. Cardiovascular diseases were more often seen in the diabetic group (16/32 in DM, 7/32 in controls, P = 0.03). Bactobilia was more often in diabetics (19 in DM, 11 in controls, p = 0.07). Postoperatively, there was a trend toward higher incidence in overall complication rate. Infectious complications (wound and respiratory infections, hepatic abscess) were higher in DM, although the difference was not statistically significant. One patient in DM group died as a result of multiorgan failure. There was no difference in total and postoperative hospital stay. The severity of acute cholecystitis was greater in diabetics (26 patients with moderate-to-severe cholecystitis in DM group, 18 in control group, P = 0.05). The study indicates that although pathological findings were more severe in DM group, the postoperative course is comparable in diabetics as compared to age and sex matched nondiabetic controls. Our findings justify reconsideration of prophylactic cholecystectomy in asymptomatic diabetic patients. PMID- 7486429 TI - "Adult" trauma surgeons with pediatric commitment: a logical solution to the pediatric trauma manpower problem. AB - Pediatric trauma care by "adult" surgeons is debated, despite the paucity of pediatric trauma surgeons; 424 patients < or = 17 admitted to a Level I Center run by "adult" surgeons were analyzed. Demographics mirrored NPTR (mean ISS 11.5; mean age 10). "Adult" critical care surgeons treated MTOS-comparable patients with outcomes comparable to MTOS. Among other specialists, only neurosurgeons saw a MTOS-comparable population. Nonoperative protocols for solid organ injury were used appropriately. Z for all patients was +0.17 with M 0.908. Ps was 0.951; acute survival was 0.958 with 18 deaths (mean Ps 0.158). There were two unexpected survivors and one unexpected death; 73% of survivors had age appropriate locomotion. Pediatric trauma outcomes by "adult" surgeons compare favorably to national standards. The recommendation that pediatric trauma care be directed by pediatric surgeons should be qualified in view of such outcomes and the paucity of pediatric trauma surgeons. PMID- 7486431 TI - Polyglyconate (Maxon) versus nylon suture in midline abdominal incision closure: a prospective randomized trial. AB - A prospective, randomized comparison of nylon versus polyglyconate (Maxon) looped suture in running mass closure of midline abdominal incisions was conducted in 225 patients. In the nylon group, 91 patients were evaluable at 2-year follow up; in the polyglyconate group, 80 patients were evaluable at follow up. Eleven patients (4 nylon and 7 polyglyconate) developed a ventral hernia. Three of the nylon and none of the polyglyconate sutured patients had a dehiscence. There was no significant difference in the overall rate of ventral hernia and dehiscence between the two suture groups. Running mass closure of midline abdominal wounds yielded satisfactory results with both of the suture materials studied. PMID- 7486430 TI - The study of the relationship between lifestyle characteristic self-reported drinking patterns and trauma. AB - A total of 161 usable interviews were obtained with 297 consecutive admissions to a Level I trauma center for purposes of evaluating patient socioeconomic patterns related to trauma. The median age of the purposeful sample was 31. There were 67.1% males, 29.2% females, and 3.7% unidentified; 43.5% were MVAs, 8.1% motorcycles, 2.5% pedestrians, 10.6% falls, and 3.7% penetrating trauma. Of those patients found ETOH positive, 40.8% admitted to drinking and driving regularly, 12.5% admitted to being previously arrested for DUI, 45% admitted that alcohol caused their current trauma, 28.6% had been previously arrested for traffic violation. Three-fourths did not routinely wear seat belts. The average education level of trauma victims was 12.2 years; 94% are blue collar workers. In summary the average trauma victim was a young adult male who had completed high school, somewhat below average earnings, and had a strong history of regular alcohol use as part of his lifestyle. PMID- 7486432 TI - Perforated jejunal diverticula: an analysis of reported cases. AB - We recently encountered two cases of perforated jejunal diverticula. We analyzed the data from all available cases of perforated jejunal diverticula, including our two cases. Our purpose was to increase awareness of this rare clinical entity as a cause of abdominal pain. An extensive literature review using Medline from its inception in 1972, and a manual review of all previously published reports was performed. Data collected included age, gender, length of symptoms from history and physical exam, operative findings, type of operation, and outcome (survival). Data analysis was performed using student's t test and multivariate analysis. Survival was not influenced by gender or type of operation. When the reports documented the number of diverticula present in the area of perforation (22 cases) a majority (19/22) had multiple diverticula. Perforated jejunal diverticula are a rare clinical entity in which age, type of operation, and gender does not influence outcome, but a longer duration of symptoms before operation trended towards a worse outcome. This clinical diagnosis should be entertained as part of any evaluation of abdominal pain. PMID- 7486434 TI - Pharyngo-esophageal perforation due to blunt trauma. AB - Pharyngoesophageal perforation due to blunt external trauma is a relatively rare and unreported injury. A patient is presented with such an injury secondary to assault, and the modes of diagnosis and management are discussed. PMID- 7486433 TI - Changing trends in management of carotid body tumors. AB - The purpose of this retrospective study was to evaluate the preoperative diagnostic modalities and aggressive operating management as well as the determination of benefit of preop embolization on the course of carotid body tumor (CBT). Between 1975 and 1993, 14 patients were treated for CBT in our department. The preoperative evaluation included angiography (14/14, 100%), Duplex scanning (6/14, 43%), and CT scanning (9/14, 64%). Five tumors (36%) were type I, four (28%) type II, and five (36%) type III, according to the Shamblin classification. In twelve cases the tumors were successfully excised without ligation of the external carotid artery. One of these patients, who had a very large tumor, underwent preop embolization. In two patients the tumors were found to extend intracranially, and the patients were referred for radiation therapy. There were no deaths or cerebrovascular complications. Temporary cranial nerve injury was noted in two cases. The pathology report revealed paragangliomas without any evidence of malignancy. Follow up of patients (6 months-18 years) showed no recurrence of the tumors in the operated cases and local recurrence in the radiation group. CONCLUSION: 1. Arteriography remains the golden standard for diagnosis of CBT. Tripplex scanning and CT are helpful for postop follow-up. 2. Surgical removal is the ideal treatment. Preoperative embolization is helpful in diminishing intraoperative blood requirements. 3. CBT, although benign, should be treated as soon as it is diagnosed, before difficulty in excision arises. PMID- 7486436 TI - Central venous catherization different for children. PMID- 7486435 TI - Rectal duplication in an adult. AB - A case is presented of a 60-year-old white female with an asymptomatic rectal duplication. The patient was initially being worked up for a vague lower abdominal pain by her gastroenterologist. Subsequently, a pelvic CT scan showed a 3 x 5 cm mass perirectally and inferior to the levator ani muscle. Surgical consultation for further evaluation and excision was arranged. A mucosal-lined cystic structure was removed and found to be consistent with rectal duplication in cystic form. PMID- 7486437 TI - Preoperative thyroid ultrasonography and fine-needle aspiration cytology in ectopic thyroid. AB - Ectopic thyroid tissue is a rare entity. The occurrence of a thyroid carcinoma in such aberrant thyroid tissue has been reported, but is very rare. Surgical excision is considered the treatment of choice in managing the lesions associated with ectopic thyroid. If the ectopic thyroid is excised, all the thyroid may have been ablated, resulting in permanent hypothyroidism. We report two cases of ectopic thyroid in emphasizing the importance of thyroid ultrasonography and fine needle aspiration (FNA) cytology in evaluating ectopic thyroid before surgery. PMID- 7486438 TI - Precholecystectomy esophagogastroduodenoscopy: is it of value? AB - Patients with cholelithiasis and history of atypical abdominal pain present the surgeon with a challenging clinical problem. We hypothesized that preoperative esophagogastroduodenoscopy before cholecystectomy would identify patients with concomitant upper gastrointestinal pathology. Retrospective review of 143 patients who presented with atypical abdominal pain, gallstones, and underwent EGD before their cholecystectomy between July 1989 and March 1994. A total of 1162 cholecystectomies were performed during the study period; 143 patients (12 per cent) underwent a preoperative EGD because of atypical abdominal pain. One hundred ten patients (77 per cent) had normal endoscopies. Thirty-three patients (23 per cent) had abnormal findings. Gastric polyps were found in three patients and esophageal varices in one patient. There were 36 findings in 29 patients, which included peptic ulcer disease, esophagitis, gastritis, and duodenitis. Seven patients had two abnormal findings. Moderate to severe disease was found in a total of 13 (9 per cent) patients. We recommend that patients who present with cholelithiasis and atypical abdominal pain undergo preoperative esophagogastroduodenoscopy, as we have found that at least 9 per cent of the patient population will have significant findings that may alter their management. PMID- 7486439 TI - Coronary pseudoaneurysm after angioplasty. AB - Coronary pseudoaneurysm is a complication of angioplasty occurring in 4-5% of cases. The likely etiology of the pseudoaneurysm is an iatrogenic dissection. Although increasingly identified, they are infrequently reported and no large series has been documented. The case of a 64-year-old patient with a post-PTCA coronary pseudoaneurysm that was treated surgically is presented. A review of the world's literature identifies stenosis as a major cofactor in operative intervention and raises the possibility that aneurysm formation promotes stenosis. PMID- 7486440 TI - Portal pyelophlebitis identified by CT scan in a patient with ischemic bowel. AB - A 65-year-old male was found to have hepatic portal venous gas (HPVG) by computed tomogram secondary to ischemic bowel. Despite aggressive surgical and antibiotic treatment, the patient died within 12 hours. Portal pyelophlebitis, when identified by air in the portal venous system, is a grave indicator of ischemic bowel with a very high mortality rate, except in patients with inflammatory bowel disease. PMID- 7486441 TI - Suppurative pylephlebitis and pylethrombosis: the role of anticoagulation. AB - Suppurative thrombophlebitis of the portal vein resulting from inflammatory intra abdominal conditions is a rare complication that may result in pylethrombosis and portal hypertension. A case is presented with documented pylethrombosis caused by diverticulitis. Color flow Doppler scanning was used to establish the diagnosis. Systemic anticoagulation therapy was added to the antibiotic regimen because of postoperative propagation of the clot. Anticoagulation therapy prompted resolution of the episode. Long-term follow-up studies demonstrated recanalization of the portal vein. Anticoagulation should be instituted with documented acute pylethrombosis caused by inflammatory disease of the abdomen. PMID- 7486442 TI - Pneumatosis intestinalis and portal venous air associated with needle catheter jejunostomy. AB - Pneumatosis intestinalis is an unusual postoperative complication. In some cases, immediate surgical intervention may be necessary. This report describes pneumatosis intestinalis with portal venous air attributed to enteral nutritional support via needle catheter jejunostomy. The etiology, radiographic findings, and management of this problem are reviewed. PMID- 7486443 TI - Antibiotic patterns associated with fungal colonization in critically ill surgical patients. AB - Fungal infections (FI) in surgical patients are increasing; mortality approaches 50 per cent. Prior studies identified factors predicting fungal colonization (FC) including broad spectrum antibiotics (BSA). This study investigates antibiotic patterns predicting FC. Other risk factors and outcomes are analyzed. A total of 72 consecutive SICU patients receiving > or = 7 days BSA were followed. None received prophylactic antifungals. Input data: Age, APACHE II, surgical procedure, lines, ulcer prophylaxis, TPN duration, antibiotic/antifungal regimen. Outcome data: FC, FI, length of SICU and hospital stay, mortality. A total of 32 patients (44%) developed FC; five (16%) developed FI (P = 0.08). All infected patients died (P = 0.0002). FC of GU (25%), respiratory (19%), and GI (19%) tracts were common. Multiple site colonization occurred in 25 per cent of colonized patients. Metronidazole and duration of ventilation predicted FC. APACHE II and TPN duration predicted mortality. Mortality occurred exclusively among patients requiring systemic antifungals. Among BSA, only metronidazole independently predisposed to FC. Other predictors of colonization and mortality agree with prior studies. The high mortality among patients requiring systemic antifungals implies that a more aggressive approach to prophylaxis may be warranted. PMID- 7486444 TI - Laparoscopic surgery for inflammatory bowel disease. AB - In the setting of inflammatory bowel disease (IBD), laparoscopic approaches have been avoided because of the often fragile intestinal tissue, thickened mesentery, malnutrition, immunosuppression, and the presence of dense adhesions. In this article, we report 10 successfully managed laparoscopic cases in IBD patients (five with ulcerative colitis, five with Crohn's Disease). Patients with ulcerative colitis underwent total abdominal colectomies, mucosal proctectomies, J-pouch construction, and diverting ileostomies. Procedures in patients with Crohn's disease included ileocecectomy (3), sigmoid colectomy with takedown of a transverse colonic fistula (1), and stricturoplasty (1). One of the 10 cases was converted to an open technique for technical reasons. Six of the 10 patients were on high dose corticosteroids for disease control. Hospital stay ranged from 6-13 days, with a median of 7 days. The morbidity rate was 20 per cent, and included one case of mild postoperative pancreatitis in a Crohn's disease patient and one delayed peri-ileostomy fistula in an ulcerative colitis patient. There was no mortality. Based on these results, we conclude that laparoscopic intestinal surgery is both feasible and safe in selected patients with inflammatory bowel disease. Use of laparoscopic techniques in these patients may reduce hospital stay, lessen adhesion formation, and improve cosmetic results in this generally young group of patients. PMID- 7486445 TI - An unsuspected cause for recurrent angina: subclavian artery stenosis. AB - The advantages of internal mammary artery (IMA) grafts over saphenous vein grafts (SVG) for coronary artery bypass grafting have been extensively recorded in the literature. Operative results and postoperative mortality in patients with IMA versus SVG are comparable. The relative risks of thrombosis, however, of an SVG are four to five times greater than an IMA graft, and the probability of recurrent angina or need for reoperation is significantly less in IMA grafted patients. Coltharp et al. also showed that the risks of reoperation for recurrent angina were decreased by a previously constructed IMA graft. PMID- 7486447 TI - Critical factors in the biology of human cancer metastasis. PMID- 7486446 TI - Methods to decrease the morbidity of abdominoperineal resection. AB - Recent reports stress that abdominoperineal resections (APR) are associated with many complications, including hemorrhage, long hospitalization, and delayed closure of an open perineal wound. Thirty-five patients underwent an abdominoperineal resection for cancer at Kaiser Permanente Medical Center, Los Angeles, from January 1989 to December 1993. All patients, except two, had their perineal wound closed after closure of the peritoneum and insertion of closed system suction catheters. Ninety-one per cent of patients achieved successful primary healing. Three patients (8.5%) suffered perineal wound dehiscence. Overall morbidity was 55 per cent, with urinary retention being the most common, occurring in 23 per cent of patients. This was managed successfully by early in and-out self catheterization. There was no incidence of urinary tract infections. There were no operative deaths. Length of stay averaged 8.6 days, with a median of 7 days. Five patients had previous radiation therapy. Of those, two (40%) had perineal wound dehiscence, compared to only one of 33 (3.3%) patients without previous radiation. APRs can be done with minimal mortality, although with an increased morbidity in irradiated patients. Primary closure and drainage of the perineal wound significantly lowers the complication rate, as does early Foley removal and self in-and-out catheterization. PMID- 7486448 TI - Inflammatory pseudotumor of the spleen: a rare entity that may mimic hematopoietic malignancy. AB - A case of benign inflammatory pseudotumor of the spleen is described in a 71-year old man who presented with left upper quadrant pain and anemia. Definitive diagnosis was made by a combination of microscopic examination with immunohistochemical stains and flow cytometry studies. To our knowledge this is only the 24th reported case of inflammatory pseudotumor involving the spleen. Recognition of this rare entity is important because the clinical manifestations and radiographic features may be indistinguishable from a malignant lymphoproliferative disorder. The clinical presentation, radiographic findings, gross and microscopic features, results of surgery, and long-term follow up are reviewed for our case and the previously reported cases in the literature. PMID- 7486449 TI - Thoracic duct injury in penetrating neck trauma. AB - Thoracic duct laceration from penetrating neck trauma is a rare injury associated with significant morbidity. Seventy-one cases published in the English literature in the last 50 years, along with one new case, were reviewed in an attempt to characterize the clinical profile, treatment, and outcome. PMID- 7486450 TI - Squamous cell carcinoma of the stomach. AB - A case of pure squamous cell carcinoma of the stomach in a 72-year-old male is reported. Epidermoid carcinomas of the stomach without a glandular cell component is a rare tumor, described in only 22 patients since 1905. It occurs mostly in males, with a male to female ratio of 5:1, affects a slightly younger age group as compared to adenocarcinoma, and has a predilection for the proximal stomach, especially along the lesser curvature. Squamous metaplasia resulting from injury to the gastric mucosa appears to have a role in the pathogenesis of this disease. The prognosis of squamous cell carcinoma of the stomach is probably less favorable than that of adenocarcinoma due to its higher frequency of lymphovascular invasion. PMID- 7486451 TI - Nonopioid analgesics shorten the duration of postoperative ileus. AB - Morphine inhibits propagating and stimulates nonpropagating colon contractions in monkeys and humans. The use of morphine or other opioids that inhibit propulsive contractions prolongs postoperative ileus. In contrast, ketorolac tromethamine, a nonsteroidal analgesic, has no effect on colon contractions in monkeys. In 14 patients having elective abdominal operations, bipolar electrodes were implanted on the right (n = 13) and left (n = 10) colon. Group A (n = 8) received ketorolac, 30 mg IM q6h, for pain relief. Group B (n = 6) needed supplemental morphine, 2-10 mg IV or IM, plus ketorolac to control their pain. Myoelectric activity was recorded from each subject on postop Days 1-5 and analyzed by computer for electrical control activity (ECA), short and long electrical response activity (ERA), and propagation of long ERA. There was a difference between the two groups in return of propagated long ERA bursts that correlated with clinical recovery from postoperative ileus. Postoperative analgesia with ketorolac resulted in faster resolution of ileus compared to morphine plus ketorolac because opioid-induced motor abnormalities in the colon were avoided. PMID- 7486452 TI - Hepatic-portal venous gas associated with ileus. AB - Hepatic portal venous gas (HPVG) is an ominous prognostic sign. There are only a few reports of survivors with this entity. More rare are cases where no intra abdominal abscess, free air, or grossly necrotic bowel exists. This a case report not only of a survivor, but of an unusual etiology that did not require surgical intervention. The inciting factor of HPVG was ileus due to anticholinergic side effects of medications and partial bowel compression from gross bladder distention. PMID- 7486453 TI - Iliopsoas abscess: etiology, diagnosis, and treatment. AB - Iliopsoas abscess is a rarely encountered entity. Early diagnosis is hindered by a nonspecific clinical presentation. The resulting delays in therapy increase morbidity and mortality rates. Our recent experience with three cases of iliopsoas abscess is presented. Based on a review of the literature, diagnostic and treatment recommendations are made. PMID- 7486454 TI - Pyloric duplications: review and case study. AB - Gastric duplications are unusual congenital anomalies that often require surgical treatment. Pyloric duplications are particularly rare; few are reported in the English literature. This article reviews the literature on pyloric duplications and describes a pyloric duplication associated with hypertrophic pyloric stenosis in a 5-week-old child and a duplication that recurred 7 years later. PMID- 7486455 TI - Malignant melanoma of the gallbladder: report of a case and review of the literature. AB - A case of primary melanoma of the gallbladder is presented and review of the literature. Eighteen cases have been previously reported. The diagnosis is usually not made preoperatively. Metastatic melanoma to the gallbladder as an isolated site is also rare. PMID- 7486456 TI - Paired temporary loop ileostomies in the localization of small bowel hemorrhage of obscure origin. AB - Hemorrhage from the gastrointestinal tract can be a challenging clinical problem when it persists and the source remains obscure. Intraoperative enteroscopy has been shown to be of value when standard diagnostic tests fail to localize the source of bleeding. We describe a case of small bowel hemorrhage that required multiple trips to the operating room. Paired ileostomies were fashioned, which led to the successful localization and resection of the responsible segment of the small intestine. PMID- 7486457 TI - Cause of temporary closure of an inner-city trauma center. AB - The number of trauma admissions, types of injury, and reasons for temporary closure were studied. Trauma patients admitted numbered 1120 in 1991, 989 in 1992, and 1164 in 1993; blunt trauma accounted for 77 percent, 74 percent, and 80 percent, whereas penetrating trauma accounted for 23 percent, 26 percent, and 20 percent of the admissions by year, respectively. A mean of 81 percent of the blunt trauma studies were for head injuries. The trauma center closed for a total of 260 hours in 1991, 211 hours in 1992, and 240 hours during 1993. Of the total hours closed, the unavailability of computed tomography (CT) scanning due to mechanical reasons was the leading cause for closure, accounting for 59 percent of hours closed. Other reasons for closure included operating (OR) unavailability (33%), acute care beds full (3.4%), blood bank stock shortage (1.4%), insufficient OR nursing teams (0.6%), unavailability of anesthesiologists (0.5%), unavailability of surgical team (1.4%), and hospital water problems (1.4%). It is concluded that designated major trauma centers may need two available CT scanners if they have a greater than 75 percent blunt trauma admission rate, or greater than 500 blunt trauma patient admissions. PMID- 7486459 TI - Charity hospital: an annotated account of the first one hundred years. PMID- 7486458 TI - Soft tissue abscesses associated with parenteral drug abuse: presentation, microbiology, and treatment. AB - Abscess formation at the site of drug injection is the commonest infectious complication in drug addicts. This study characterizes the clinical presentation of the condition, its current microbiology, and treatment outcome. All patients presenting for treatment of soft tissue abscesses associated with parenteral drug abuse over a 21-month period were studied. Sixty-six patients with 70 subcutaneous abscesses after injection of cocaine (85%), heroin (5%), or unreported drugs (10%) were identified. Only 42% were febrile (T > 37.5 degrees C), 54 percent had leukocytosis, and 47 percent had wound fluctuance. Wound cultures (243 isolates in 57 patients) grew predominately anaerobes (143 isolates) and facultative gram-positive cocci (88 isolates). Twenty-six blood cultures were obtained, and five (19%) were positive, two with the same bacteria isolated from the wound. Of the patients tested, 29 percent were positive for hepatitis B surface antigen and 9 percent for HIV. Simple incision and drainage was effective in all cases. Classical signs and symptoms of infection and abscess formation may be absent in this patient population. Many of these patients carry other blood-borne infections which the health professional must guard against. Cocaine injection, and "mixed" aerobic-anaerobic infections predominated, in contrast to earlier reports, when narcotics and aerobes predominated. Simple incision and drainage is adequate treatment; antibiotics, when given, should cover gram-positive and anaerobic bacteria; gram-negative coverage is unnecessary. PMID- 7486460 TI - The role of the Opaque2 transcriptional factor in the regulation of protein accumulation and amino acid metabolism in maize seeds. PMID- 7486461 TI - Age-dependent doses to members of the public from intake of radionuclides: Part 3. Ingestion dose coefficients. A report of a Task Group of Committee 2 of the International Commission on Radiological Protection. PMID- 7486462 TI - Accuracy of transesophageal echocardiography for identifying left atrial thrombi. A prospective, intraoperative study. AB - OBJECTIVE: To determine the ability of transesophageal echocardiography to accurately identify or exclude left atrial thrombi. DESIGN: Prospective cohort study. SETTING: University hospital. PATIENTS: 231 consecutive patients having transesophageal echocardiography before elective repair or replacement of the mitral valve or excision of a left atrial tumor. Fifty-six percent of patients had a history of atrial fibrillation, and 17% had a history of thromboembolism. MEASUREMENT: Identification of left atrial thrombi during transesophageal echocardiographic examination and comparison with direct near-simultaneous visualization during cardiac surgery. RESULTS: Transesophageal echocardiography identified 14 left atrial thrombi in 14 patients (6%). Thrombus size range from 3 to 80 mm. Surgery confirmed 12 of 14 thrombi (86%), including 9 thrombi confined to the left appendage. No additional thrombi were found on direct inspection of the atria (sensitivity, 100% [95% CI, 74% to 100%]; specificity, 99% [CI, 97% to 99.9%]; positive predictive value, 86% [12/14]; negative predictive value, 100% [217/217]; for a population that had a 5.2% prevalence of thrombi). All 12 surgically confirmed thrombi were identified by two independent observers. Neither thrombus seen by only a single observer on transesophageal echocardiography was confirmed during direct inspection of the atria at surgery. CONCLUSION: Transesophageal echocardiography is highly accurate for identifying left atrial thrombi and can be used clinically to exclude left atrial thrombi. PMID- 7486463 TI - Mortality in patients with hemophilia. Changes in a Dutch population from 1986 to 1992 and 1973 to 1986. AB - OBJECTIVE: To determine causes of death and mortality rates in patients with hemophilia over a period of 20 years, to assess changes in mortality, and to distinguish between hemophilia-related death and recent death induced by viral infections. DESIGN: Cohort study of 919 patients followed from January 1986 to June 1992. Results were compared with outcomes of previous follow-up from 1973 to 1986. SETTING: Consecutive national questionnaire surveys on hemophilia, using patient registries of the Netherlands Hemophilia Society and Dutch hemophilia centers. PATIENTS: 919 males hemophilia A or B who participated in a national questionnaire survey on hemophilia in 1985. Median duration of follow-up was 6.4 years, which yielded 5753 person-years of follow-up. The mean age at study entry was 30 years (range, 1 to 85 years). MEASUREMENTS: Standardized mortality ratios, causes of death, median life expectancy, age-adjusted relative risks associated with the type or severity of hemophilia, presence of inhibitors, prophylaxis, and human immunodeficiency virus infection. RESULTS: 45 patients (5%) died between January 1986 and June 1992; 22.6 patients had been expected to die. Thus, the overall standardized mortality ratio was 2.0. The overall median life expectancy was 66 years for the cohort studied from 1973 to 1986 and 68 years for the cohort studied from 1986 to 1992. When deaths related to viral infection were excluded, the life expectancy almost equaled that of the general male population. Between 1986 and 1992, 1 patient died of ischemic heart disease compared with the 5.2 who were expected to die of this disease. Infection with HIV was the strongest independent predictor of death (relative risk, 27.5 [95% CI, 5.7 to 132.8]). After adjustment for HIV infection, no other hemophilia-related risk factors were associated with the risk of death. CONCLUSIONS: The acquired immunodeficiency syndrome and hepatitis strongly influence mortality in patients with hemophilia. In the absence of viral infections, the life expectancy of patients with hemophilia would almost equal that of the general male population. PMID- 7486464 TI - Antibody responses to polysaccharide and polysaccharide-conjugate vaccines after treatment of Hodgkin disease. AB - OBJECTIVE: To compare the immunogenicity of polysaccharide-conjugate vaccines with that of polysaccharide vaccines in patients previously treated for Hodgkin disease. DESIGN: All patients were immunized with Haemophilus influenzae type b (HIB)-conjugate and 4-valent meningococcal polysaccharide vaccines. Subgroups of patients were randomly assigned to receive either 23-valent pneumococcal polysaccharide vaccine or a 7-valent pneumococcal-conjugate vaccine that links seven pneumococcal serotypes to the outer membrane protein complex of Neisseria meningitidis. PATIENTS: 144 patients who had completed treatment for Hodgkin disease, which had been diagnosed at least 2 years before the study. MEASUREMENTS: Antigen-specific antibody concentrations before and 3 to 6 weeks after immunization; number of persons who achieved anti-HIB antibody concentrations considered to be in the protective range. RESULTS: The geometric mean anti-HIB antibody concentration increased from 1.79 micrograms/mL before immunization to 54.1 micrograms/mL after; the percentage of persons with antibody concentrations in the protective range increased from 62% before immunization to 99% after. Patients immunized with 23-valent pneumococcal vaccine had a geometric mean pneumococcal antibody concentration after immunization (9.15 micrograms/mL) that was similar to that of healthy controls (10.0 micrograms/mL) for the seven serotypes measured. In contrast, patients who received 7-valent pneumococcal conjugate vaccine had a significantly lower mean response compared with patients who received 23-valent; their geometric mean antibody concentration after immunization was 4.95 micrograms/mL (P = 0.005). CONCLUSION: A single dose of HIB conjugate vaccine was immunogenic in patients who had completed treatment for Hodgkin disease diagnosed at least 2 years before immunization. In addition, responses to the 23-valent pneumococcal and 4-valent meningococcal vaccines were equivalent to those seen in healthy controls. Finally, patients had a significantly lower response to a single dose of 7-valent pneumococcal-conjugate vaccine than to 23-valent vaccine. PMID- 7486465 TI - Clostridium difficile colitis: an efficient clinical approach to diagnosis. AB - OBJECTIVE: To define clinical and laboratory variables that suggest the presence of Clostridium difficile colitis and to establish the number of stool specimens needed to reasonably exclude the diagnosis of C. difficile colitis. DESIGN: Prospective study of consecutive inpatients whose stool specimens were sent to be evaluated for the presence of C. difficile toxin. SETTING: University teaching hospital. PATIENTS: 268 hospital inpatients in medical, surgical, and gynecology units. MEASUREMENTS: Structured history and physical examination; detection of C. difficile toxin by cytotoxin tissue-culture assay with anti-C. difficile antiserum neutralization and by enzyme-linked immunoassay (EIA) for C. difficile toxins A and B; and detection of fecal leukocytes by microscopic examination and by latex agglutination lactoferrin assay. RESULTS: 43 of 268 consecutive inpatients were positive for C. difficile toxin by EIA or tissue-culture assay. Although toxin was detected by EIA alone in 39 of the 43 patients, it was detected in an additional 4 patients (10%) by tissue-culture assay alone. Univariate and multivariate logistic regression analysis showed that the following clinical and laboratory features were associated with C. difficile toxin positivity: the onset of diarrhea 6 or more days after the administration of antibiotics (odds ratio, 1.38 [95% CI, 1.10 to 3.79]); hospital stay longer than 15 days (odds ratio, 1.33 [CI, 1.09 to 3.95]); the presence of fecal leukocytes determined by microscopy (odds ratio, 2.39 [CI, 1.05 to 5.42]) or lactoferrin assay (odds ratio, 3.74 [CI, 1.80 to 7.76]); the presence of semiformed (as opposed to watery) stools (odds ratio, 2.33 [CI, 1.10 to 4.90]); and cephalosporin use (odds ratio, 2.36 [CI, 1.10 to 5.09]). Toxin-positive patients were no more likely than controls to have had fever, abdominal pain or cramps, leukocytosis, green-colored diarrhea, or blood in the stool or to have received clindamycin or penicillin derivatives. Of the 43 patients with C. difficile toxin, 34 (79%) had positive results for the toxin on the first stool specimen, 5 (cumulative, 91%) had positive results on the second specimen, and 4 had positive results on the third specimen. Overall, the negative predictive value of the first stool specimen was 97%. All patients who had two or more clinical or laboratory predictors were diagnosed with C. difficile disease when either the first or the second stool specimen was positive for toxin. CONCLUSIONS: Clinicians at the bedside can use readily available clinical and laboratory information to decide which patients are likely to have C. difficile disease and when it is appropriate and useful to order specific diagnostic tests for C. difficile toxin. Such data are also useful in determining the number of stool samples that reasonably excludes the diagnosis of C. difficile colitis. PMID- 7486466 TI - The risk of midline catheterization in hospitalized patients. A prospective study. AB - OBJECTIVE: To assess the risk associated with midline catheter use in hospitalized patients. DESIGN: Prospective, consecutive enrollment. SETTING: A 719-bed university-affiliated hospital. PATIENTS: Patients were enrolled if they were likely to require at least 7 days of intravascular catheterization while hospitalized. MEASUREMENTS: Patients were monitored for adverse reactions. Catheter segment, insertion site, hub, infusate, and blood cultures were assessed. RESULTS: From February 1993 through June 1994, 251 Landmark midline catheters were inserted in 238 patients. One hundred forty catheter cultures were obtained from 130 patients who remained hospitalized for the duration of catheterization. For these 130 patients, the mean duration of catheterization was 9 days, the incidence of catheter colonization was 5.0 per 1000 catheter days, and the incidence of catheter-related bloodstream infection was 0.8 per 1000 catheter days. During the study period, two severe, unexpected adverse reactions occurred that may have been associated with the use of Landmark midline catheters; no such reactions were associated with the insertion of 58,580 Teflon peripheral catheters (P < 0.00001; exact 95% lower bound of the odds ratio, 68.9). Fifty-three similar reactions associated with Landmark midline catheters, including two deaths, have been reported to the Food and Drug Administration through June 1994. CONCLUSION: The risk for midline catheter-related infection is low. However, Landmark midline catheters are associated with life-threatening adverse reactions that are probably attributable to the catheter material itself. PMID- 7486467 TI - Increased genital shedding of herpes simplex virus type 2 in HIV-seropositive women. AB - OBJECTIVE: To compare the prevalence of genital herpes simplex virus type 2 (HSV 2) shedding in human immunodeficiency virus (HIV)-seropositive women and HIV seronegative women. DESIGN: Cross-sectional study. SETTING: A major inner-city medical center. PATIENTS: 106 women who were HIV-seropositive and HSV-2 seropositive and 70 women who were HIV-seronegative and HSV-2-seropositive were enrolled from various primary care settings. MEASUREMENTS: Herpes simplex virus type 2 antibody determinations were done for all patients. Regardless of symptoms, vulvar and cervical HSV cultures were obtained from all HIV seropositive women and from a randomly selected subgroup of HIV-seronegative women. RESULTS: The prevalence of HSV-2 shedding was nearly four times greater in HIV-seropositive than in HIV-seronegative women (13.2% compared with 3.6%; P = 0.04; odds ratio, 4.1 [95% CI, 1.0 to 27.4]) when the serum antibody for HSV-2 was present. Seventy-nine percent of viral shedding among HIV-seropositive women was asymptomatic. Overall viral shedding increased significantly as the CD4 cell count decreased. CONCLUSIONS: Women with HIV infection, particularly those with low CD4 cell counts, shed HSV-2 from the vulva and cervix more commonly than women not infected with HIV. Most of this shedding is asymptomatic. PMID- 7486468 TI - Transcranial Doppler ultrasonography during head-upright tilt-table testing. PMID- 7486469 TI - Nutritional issues in nursing home care. AB - The most common nutritional problems in nursing home residents are weight loss and concomitant protein energy undernutrition. Although the causes of weight loss in these patients can usually be treated, they are rarely identified in the nursing home. Depression and adverse drug effects are the most common causes of weight loss. We discuss the appropriate use of feeding tubes in the nursing home and the early use of enteral feeding to prevent the development of severe protein energy undernutrition. Vitamin deficiencies, especially folate and pyridoxine deficiencies, frequently develop in nursing home residents. Hip fractures are often associated with vitamin D deficiency. Trace mineral deficiencies (for example, zinc deficiency) can aggravate immune deficiency and slow wound healing. Inadequate fluid intake leads to dehydration, hypotension, and, in persons with diabetes mellitus, hyperosmolarity. Finally, food intake itself can cause postprandial hypotension (which in turn may precipitate falls), produce electrolyte shifts, and result in aspiration pneumonia. Physical activity programs are an important component of nursing home care that may have an effect on nutritional status, and simple, cost-effective programs may be as beneficial as high-technology programs. Careful attention to the nutritional intake of nursing home residents is both a clinical and a quality-of-life issue. PMID- 7486470 TI - The antioxidant vitamins and cardiovascular disease. A critical review of epidemiologic and clinical trial data. AB - PURPOSE: To review prospective epidemiologic studies and randomized trials regarding the role of antioxidant vitamins (vitamins E and C and beta-carotene) in the prevention of cardiovascular disease, with emphasis on differences in results obtained by these two types of studies. DATA SOURCES: Computerized and manual searches of the literature on antioxidant vitamins and cardiovascular disease. STUDY SELECTION: Prospective epidemiologic studies and randomized trials that included 100 or more participants and provided quantified estimates of antioxidant vitamin intake. DATA SYNTHESIS: Comparisons of relative risk reductions (RRR) across observational studies and randomized trials, including assessment of dose-response relations. RESULTS: All three large epidemiologic cohort studies of vitamin E noted that high-level vitamin E intake or supplementation was associated with a significant reduction in cardiovascular disease (RRR range, 31% to 65%), as measured by various fatal and nonfatal cardiovascular end points. To obtain these reductions, vitamin E supplementation must last at least 2 years. Less consistent reductions were seen in studies of beta-carotene (RRR range, -2% to 46%) and vitamin C (RRR range, -25% to 51%). Considerable biases in observational studies, such as different health behaviors of persons using antioxidants, may account for the observed benefit. By contrast, none of the completed randomized trials showed any clear reduction in cardiovascular disease with vitamin E, vitamin C, or beta-carotene supplementation. The trials were not specifically designed to assess cardiovascular disease, did not provide data on nonfatal cardiovascular end points, may have had insufficient treatment durations, and used suboptimal vitamin E doses. The completed trials were of adequate size to indicate that the true therapeutic benefit of vitamin E and other antioxidants in reducing fatal cardiovascular disease (a survival benefit as long as 5 years) is probably more modest than the epidemiologic data suggest. CONCLUSION: The epidemiologic data suggest that antioxidant vitamins reduce cardiovascular disease, with the clearest effect for vitamin E; however, completed randomized trials do not support this finding. Much of this controversy should be resolved by the ongoing large-scale and long-term randomized trials designed specifically to evaluate effects on cardiovascular disease. PMID- 7486471 TI - Discordance between meta-analyses and large-scale randomized, controlled trials. Examples from the management of acute myocardial infarction. AB - Clinicians making treatment decisions are faced with ever-growing numbers of therapies, each supported by different types of clinical data. By bringing together large amounts of data, meta-analysis has emerged as a useful tool for generating hypotheses with which to plan definitive trials, and it has also been recommended as a basis for decision making in the absence of definitive trials. In several instances, early meta-analyses have provided evidence of efficacy that was subsequently confirmed. However, in other instances, the results of initial meta-analyses have disagreed with the results of subsequent large-scale trials. Nitrate and magnesium therapy for acute myocardial infarction are two contemporary examples of treatments about which hypothesis-generating meta analyses and subsequent large trials have disagreed. We review the issues surrounding the interpretation of meta-analyses in these cases, and we suggest that the appropriate use of meta-analyses in clinical decision making be carefully placed in the context of a review of pathophysiologic principles and the results of basic laboratory research and individual trials. PMID- 7486472 TI - Competent patients, incompetent decisions. AB - We describe the epidemiologic aspects of dialysis withdrawal, the role of patient autonomy and of medical beneficence in the decision to discontinue dialysis, and the conditions under which limits may be placed on patient autonomy. A case is presented that provides insight into the issues involved in making such decisions and that should provoke further discussion on the role of "medical guidelines" for dialysis refusal and withdrawal. PMID- 7486473 TI - Shadows on the cave wall: the role of transesophageal echocardiography in atrial fibrillation. PMID- 7486474 TI - Reactions associated with midline catheters for intravenous access. PMID- 7486475 TI - Antioxidants and cardiovascular disease: why do we still not have the answers? PMID- 7486477 TI - Post-midnight reverie. PMID- 7486476 TI - Case-control study, meta-analysis, and bouillabaisse: putting the calcium antagonist scare into context. PMID- 7486478 TI - Do diuretics cause heart disease? PMID- 7486479 TI - Weight loss and longevity. PMID- 7486480 TI - More on patients with implantable cardioverter-defibrillators. PMID- 7486481 TI - Masked hyperprolactinemia in a case of aldosterone-producing adrenal adenoma. PMID- 7486482 TI - Cutaneous vasculitis associated with mefloquine. PMID- 7486484 TI - Prevention of colorectal cancer by flexible endoscopy and polypectomy. A case control study of 32,702 veterans. AB - OBJECTIVE: To determine whether patients with colorectal cancer are less likely than unaffected controls to have had one or more endoscopic procedures (flexible sigmoidoscopy, colonoscopy, or polypectomy) before being diagnosed with cancer. DESIGN: Case-control study. SETTING: Hospitals of the Department of Veterans Affairs. PATIENTS: 8722 and 7629 case-patients with colon and rectal cancer, respectively, and age-, sex-, race-matched controls who were discharged at the same time as the corresponding case-patients. MEASUREMENTS: Number and type of endoscopic procedures of the large bowel done from 1981 until the development of colorectal cancer in each case-patient. The influence of endoscopic procedures on the development of colorectal cancer was tested by conditional multiple logistic regression analysis. RESULTS: Compared with controls, patients with colorectal cancer were less likely to have had an endoscopic procedure of the large bowel before being diagnosed with cancer (odds ratio for colon cancer, 0.51 [95% CI, 0.44 to 0.58]; odds ratio for rectal cancer, 0.55 [CI, 0.47 to 0.64]). In patients who had flexible sigmoidoscopy, colonoscopy, and polypectomy, the odds ratios were even smaller. When analyzed by separate 1-year intervals, patients with cancer had significantly fewer procedures during periods of up to 6 years before their onset of their cancer. Similarly, fewer inpatient and outpatient procedures were done in patients than in controls. CONCLUSIONS: Endoscopic procedures of the large bowel reduce the risk for developing colon and rectal cancer by 50%, their protective influence lasting 6 years. PMID- 7486483 TI - Ribavirin as therapy for chronic hepatitis C. A randomized, double-blind, placebo controlled trial. AB - OBJECTIVE: To evaluate ribavirin, an oral antiviral agent, as therapy for chronic hepatitis C. DESIGN: Randomized, double-blind, placebo-controlled study. SETTING: Clinical Center of the National Institutes of Health, a tertiary referral research hospital. PATIENTS: 29 patients with chronic hepatitis C who received oral ribavirin (600 mg twice daily) for 12 months and 29 controls with chronic hepatitis C who received placebo for 12 months. MEASUREMENTS: Effects of therapy were evaluated by measuring serum aminotransferase and hepatitis C virus (HCV) RNA levels before, during, and for 6 months after therapy and by histologic examination of liver specimens before and at the end of treatment. RESULTS: Patients treated with ribavirin had a prompt decrease in serum aminotransferase levels (54% overall) compared with levels before treatment and levels in controls (5% decrease). Serum aminotransferase levels became normal or nearly normal in 10 patients treated with ribavirin (35% [95% CI, 18% to 54%]) but in no controls (0% [CI, 0% to 12%]). Aminotransferase levels remained normal in only 2 patients after ribavirin therapy was discontinued (7% [CI, 1% to 23%]). Serum HCV RNA levels did not change during or after therapy. Liver biopsy specimens showed a decrease in hepatic inflammation and necrosis among ribavirin-treated patients whose aminotransferase levels became normal. CONCLUSIONS: Ribavirin has beneficial effects on serum aminotransferase levels and histologic findings in the liver in patients with chronic hepatitis C, but these effects are not accompanied by changes in HCV RNA levels and are not sustained when ribavirin therapy is discontinued. Thus, ribavirin alone for periods as long as 12 months is unlikely to be of value as therapy for chronic hepatitis C. PMID- 7486485 TI - Effects of n-3 polyunsaturated fatty acids on glucose homeostasis and blood pressure in essential hypertension. A randomized, controlled trial. AB - OBJECTIVE: To determine whether dietary supplementation with fish oil adversely affects glycemic control in patients with hypertension. DESIGN: Randomized, double-blind, placebo-controlled study. PATIENTS: 78 persons with untreated hypertension recruited from a population survey. INTERVENTION: Participants were randomly assigned to receive eicosapentaenoic and docosahexaenoic acids, 4 g/d, or corn oil placebo, 4 g/d, for 16 weeks. MEASUREMENTS: An oral glucose tolerance test; assessments of insulin release, glucose disposal, and insulin sensitivity done using the hyperglycemic clamp technique to keep plasma glucose levels at 10 mmol/L for 180 minutes; assessment of insulin sensitivity done using a euglycemic hyperinsulinemic clamp technique (infusing insulin and glucose to keep plasma glucose levels at 5 mmol/L); assessments of lipid levels and blood pressure. Measurements were done before and after intervention. RESULTS: Changes in integrated glucose and insulin response after the oral glucose challenge did not differ between the fish oil and corn oil groups after intervention (-0.6 +/- 0.7 compared with -1.0 +/- 0.6 mmol/L [P > 0.3] for integrated glucose and 143 +/- 76 compared with 169 +/- 84 pmol/L [P > 0.3] for insulin response). Changes in first phase insulin release (34 +/- 72 pmol/L in the fish oil group compared with 191 +/- 112 pmol/L in the corn oil group [P > 0.3]), second-phase insulin release (179 +/- 66 pmol/L compared with 257 +/- 122 pmol/L [P > 0.3]), and insulin sensitivity index (-0.03 +/- 0.01 compared with -0.01 +/- 0.01 [mumol/kg.min divided by pmol/L]; P > 0.3) were also similar in both groups after treatment. Fish oil lowered systolic blood pressure by 3.8 mm Hg more than control (P = 0.04) and lowered diastolic blood pressure by 2.0 mm Hg more than control (P = 0.10). After fish oil treatment, triglyceride levels decreased by 0.28 +/- 0.08 mmol/L more than control (P = 0.01), and very-low-density lipoprotein cholesterol levels decreased by 0.13 +/- 0.04 mmol/L more than control (P = 0.01). CONCLUSION: Fish oil, in doses that reduce blood pressure and lipid levels in hypertensive persons, does not adversely affect glucose metabolism. PMID- 7486486 TI - Polymorphonuclear leukocytes in non-insulin-dependent diabetes mellitus: abnormalities in metabolism and function. AB - OBJECTIVE: To determine basal levels of cytosolic calcium ([Ca2+]i) and phagocytic activity in polymorphonuclear leukocytes (PMNLs) from patients with non-insulin-dependent diabetes (NIDDM). DESIGN: Prospective cohort study. SETTING: A university-county hospital. MEASUREMENTS: Cytosolic calcium levels, adenosine triphosphate (ATP) content, and phagocytosis of PMNLs from patients with NIDDM and from controls. INTERVENTION: In patients with NIDDM, we evaluated the effect of treatment with an oral hypoglycemic agent (glyburide) on [Ca2+]i levels, ATP content, and the phagocytosis of PMNLs. PATIENTS: 22 controls and 34 patients with NIDDM were examined. Fifteen patients were studied before and after 3 months of treatment with glyburide. RESULTS: Polymorphonuclear leukocytes from patients with NIDDM showed significantly elevated basal levels of [Ca2+]i (68 +/- 9.6 compared with 43 +/- 4.9 nmol/L; P < 0.01); reduced ATP content (1.30 +/- 0.58 compared with 2.35 +/- 0.45 nmol/10(6) PMNLs; P < 0.01); and impaired phagocytosis (117 +/- 21.0 compared with 145 +/- 17.4 micrograms oil/10(7) PMNLs per minute; P < 0.01) compared with controls. There was a direct and significant correlation (P < 0.01, r = 0.80) between [Ca2+]i levels in PMNLs and serum glucose levels and an inverse correlation between phagocytic ability and [Ca2+]i levels (P < 0.01; r = 0.62) as well as between phagocytic activity and fasting serum glucose levels (P < 0.01, r = 0.54) in patients with NIDDM. Glyburide therapy resulted in significant reduction in fasting serum glucose levels; in PMNLs, this treatment resulted in a significant reduction in [Ca2+]i levels, a significant increase in ATP content, and a significant improvement of phagocytosis. CONCLUSIONS: Patients with NIDDM have elevated [Ca2+]i levels in PMNLs. This abnormality is probably induced by hyperglycemia and is primarily responsible for the imparied phagocytosis seen in these patients. PMID- 7486487 TI - The role of antineutrophil cytoplasmic antibody (c-ANCA) testing in the diagnosis of Wegener granulomatosis. A literature review and meta-analysis. AB - PURPOSE: To summarize the literature on the clinical utility of antineutrophil cytoplasmic antibody (c-ANCA) as a diagnostic marker for Wegener granulomatosis. DATA SOURCES: A structured literature search was done using MEDLINE; this search, combined with a manual search, yielded 747 articles for potential inclusion. These articles passed through a 4-stage review. Studies were included if they had a specified reference standard and a systematic method of patient selection and if a 2 x 2 contingency table could be constructed from their results. Fifteen articles met these criteria. DATA EXTRACTION: 3 physicians reviewed all selected articles. Detailed data abstraction was done, and the quality of the study methods was assessed. Items abstracted included the method of patient selection, the study design, the reference standard, the c-ANCA testing technique, disease activity, and the test results. Disagreements among reviewers were resolved by consensus. Contingency tables were used to calculate the operating characteristics for Wegener granulomatosis overall and for active and inactive Wegener granulomatosis. RESULTS: The sensitivities of c-ANCA testing for overall Wegener granulomatosis ranged from 34% to 92%, and the specificities ranged from 88% to 100%. The pooled sensitivity was 66% (95% CI, 57% to 74%), and the pooled specificity was 98% (CI, 96% to 99.5%). Four articles provided data on disease activity. For active disease, the pooled sensitivity was 91% (CI, 87% to 95%), and the pooled specificity was 99% (CI, 97% to 99.9%). For inactive disease, the pooled sensitivity and specificity were 63% and 99.5%, respectively. CONCLUSIONS: Although c-ANCA test results may serve clinicians as adjunct evidence for the diagnosis of Wegener granulomatosis, these results must be viewed in the context of the patient's clinical picture and disease activity and the prevalence of Wegener granulomatosis in the clinical setting in which the patient is seen. PMID- 7486488 TI - Trends in infectious diseases and cancers among persons dying of HIV infection in the United States from 1987 to 1992. AB - OBJECTIVE: To determine trends in the relative frequency of infectious diseases and cancers among U.S. residents dying of human immunodeficiency virus (HIV) infection. DATA SOURCE: National multiple-cause mortality data for 1987 to 1992 compiled from death certificates. SUBJECTS: Deaths reported with HIV infection as the underlying cause and with nonunderlying causes that could be secondary to HIV infection. DATA ANALYSIS: Trends in the annual percentage of deaths associated with each infectious disease or cancer that accounted for at least 1.0% of all HIV-related deaths. RESULTS: From 1987 to 1992, the percentage of HIV-related deaths associated with the following diseases decreased: pneumocystosis, from 32.5% to 13.8%; cryptococcosis, from 7.7% to 5.0%; and candidiasis, from 2.3% to 1.7%. The percentage of deaths associated with the following diseases increased: nontuberculous mycobacteriosis, from 6.7% to 12.2%; cytomegalovirus disease, from 5.2% to 9.9%; bacterial septicemia, from 9.0% to 11.5%; non-Hodgkin lymphoma, from 3.9% to 5.7%; tuberculosis, from 2.9% to 4.1%; progressive multifocal leukoencephalopathy, from 0.8% to 1.9%; bacterial pneumonia, from 1.2% to 2.1%; and cryptosporidiosis or isosporiasis, from 0.7% to 1.2%. The percentages of deaths associated with toxoplasmosis, Kaposi sarcoma, and pneumonia caused by unspecified organisms had no significant linear trends (ranges from 4.9% to 5.5%, 10.4% to 12.1%, and 17.6% to 18.6%, respectively). CONCLUSIONS: The percentage of HIV-related deaths associated with pneumocystosis has decreased dramatically, probably because of chemoprophylaxis and improved treatment. Pneumonia caused by unspecified organisms has now become the leading secondary cause of death among persons dying of HIV infection. Decreases in the percentages of HIV-related deaths associated with cryptococcosis and candidiasis may reflect the use of new antifungal agents such as fluconazole. PMID- 7486490 TI - Critical pathways as a strategy for improving care: problems and potential. AB - In an era of increasing competition in medical care, critical pathway guidelines have emerged as one of the most popular new initiatives intended to reduce costs while maintaining or even improving the quality of care. Developed primarily for high-volume hospital diagnoses, critical pathways display goals for patients and provide the corresponding ideal sequence and timing of staff actions for achieving those goals with optimal efficiency. Despite the rapid dissemination of critical pathway programs in hospitals throughout the United States, many uncertainties remain about their development, implementation, and evaluation. In addition, serious concerns have been raised about their effect on patient outcomes and satisfaction with care, physician autonomy, malpractice risks, and the teaching and research missions of many hospitals. Underlying these concerns is the absence of data from controlled trials to evaluate the effects of critical pathways. Physicians should understand the potential benefits and problems associated with critical pathways because physicians are increasingly being asked to provide leadership for pathway programs. Physicians and other health service investigators should also develop methods to study pathways in evolving health care settings. Although the promise of reduced costs and improved quality is enticing, the gaps in our knowledge about critical pathways are extensive; therefore, like any new health care technology, pathway programs should be fully evaluated in order to understand the conditions under which that promise may be fulfilled. PMID- 7486491 TI - Prevention of colorectal cancer by endoscopic polypectomy. PMID- 7486489 TI - Sporadic meningococcal disease in adults: results of a 5-year population-based study. AB - OBJECTIVE: To define the incidence, demographics, clinical features, and risk factors for sporadic meningococcal disease in adults (> or = 18 years) residing in metropolitan Atlanta. DESIGN: Prospective, population-based surveillance, with retrospective review of clinical and laboratory records. SETTING: Eight-county metropolitan Atlanta area. PATIENTS: All adult patients in whom Neisseria meningitidis was isolated from normally sterile sites (blood, cerebrospinal fluid) during the period 1 December 1988 to 30 November 1993. MEASUREMENTS: Incidence, relative risk, clinical and laboratory parameters, and serogroup of meningococcal isolates. RESULTS: For the 5-year period, 44 (33%) of 132 cases of meningococcal disease in Atlanta occurred in adults (annual incidence, 0.50/100,000 adults per year). Twenty-three (52%) of the 44 adults presented without rash or meningitis, the two most obvious signs of meningococcal disease. Pneumonia, sinusitis, or purulent tracheobronchitis, but without rash, were the likely sources of meningococcal bacteremia in 15 (34%) of the 44 adults. Twelve of the 15 patients with meningococcal respiratory infection were older than 50 years of age or were immunocompromised (or both), and three fourths of the 15 patients had disease caused by serogroups B, Y, and W-135. Overall, two thirds of adults older than 24 years of age with meningococcal disease had one or more immunocompromising conditions (for example, low complement 50 level [CH50], corticosteroid use, congestive heart failure, multiple myeloma, human immunodeficiency virus infection). Meningococcemia or meningococcal meningitis, often caused by serogroup C, were the presentations in 14 of 15 adults 18 to 24 years old; only 2 had an identified underlying condition. CONCLUSIONS: In this 5 year population-based study, one third of all cases of sporadic meningococcal disease occurred in adults. Over half of the adults presented without rash or meningitis. Pneumonia, sinusitis, and tracheobronchitis are important sources of bacteremic meningococcal disease, especially in immunocompromised patients and elderly persons. PMID- 7486492 TI - Diabetes, fish oil, and vascular disease. PMID- 7486495 TI - Marrying medicine. PMID- 7486493 TI - Normalization of intracellular calcium: a sweet solution to neutrophil dysfunction in diabetes? PMID- 7486494 TI - . . . and now microsporidiosis. PMID- 7486496 TI - Postmenopausal hormone therapy and systemic lupus erythematosus. PMID- 7486497 TI - Postmenopausal hormone therapy and systemic lupus erythematosus. PMID- 7486498 TI - The repetitive short corticotropin stimulation test in patients with septic shock. PMID- 7486499 TI - The exodus of a medical school. PMID- 7486501 TI - The exodus of a medical school. PMID- 7486502 TI - Medicine and the Holocaust. PMID- 7486500 TI - The exodus of a medical school. PMID- 7486503 TI - Medicine and the Holocaust. PMID- 7486504 TI - Literature and medicine: contributions to clinical practice. PMID- 7486505 TI - Literature and medicine: contributions to clinical practice. PMID- 7486506 TI - Literature and medicine: contributions to clinical practice. PMID- 7486507 TI - Human B-cell development. PMID- 7486508 TI - Regulation of N-region diversity in Ig heavy chain transcripts expressed by murine fetal liver cells cultured with IL-7. PMID- 7486509 TI - Mutations in immunoglobulin V gene promoters may cause reduced germline transcription and diminished recombination frequencies. PMID- 7486510 TI - High frequency of normal IgH rearrangement in scid mice. PMID- 7486512 TI - Human immunoglobulin isotype switching. Studies at the DNA level. PMID- 7486513 TI - Role of Oct-2 in immunoglobulin gene silencing. PMID- 7486511 TI - Regulation of antibody class switching to IgE: characterization of an IL-4 responsive region in the immunoglobulin heavy-chain germline epsilon promoter. AB - A large body of data indicate that antibody class switching is directed by cytokines by inducing or repressing transcription from unrearranged, or germline, CH genes. IL-4 induces transcription of the germline C epsilon genes in activated B cells, and subsequently cells in this population will undergo switch recombination to IgE. Furthermore, the data suggest that transcription of germline C epsilon genes is required for class switching. In this paper we define DNA elements required for induction of transcription of the germline C epsilon genes by IL-4. To do this, segments of DNA from the 5' flank of the initiation sites for germline epsilon RNA were ligated to a luciferase reporter gene and transfected into two mouse B-cell lines, one of which can be induced to switch to IgE. By analysis of a series of 5' deletion constructs and linker-scanning mutations, we demonstrate that a 46-bp segment (residing at -126/-79 relative to the first RNA initiation site) contains an IL-4 responsive region. This segment binds three transcription factors: the recently described NF-IL4, one or more members of the C/EBP family of transcription factors, and NF-kappa B/p50. Mutation of any of the binding sites for these three factors abolishes or reduces IL-4 inducibility of the epsilon promoter. A 27-bp segment within this IL-4 response region containing binding sites for NF-IL4 and a C/EBP factor is sufficient to transfer IL-4 inducibility to a minimal c-fos promoter. PMID- 7486514 TI - Molecular evidence that in vivo isotype switching occurs within the germinal centers. PMID- 7486515 TI - Diversification of bovine lambda-light chain genes. PMID- 7486517 TI - A sequence analysis of human germline Ig VH and VL genes. The CDR1s of a major proportion of VH, but not VL, genes display a high inherent susceptibility to amino acid replacement. PMID- 7486516 TI - Somatic mutation of human immunoglobulin V genes: bias, rate, and regulation. PMID- 7486518 TI - Comparison of the human germline and rearranged VH repertoire reveals complementarity between germline variability and somatic mutation. PMID- 7486519 TI - Analysis of somatic mutation with a reporter gene construct. PMID- 7486520 TI - Mechanistic properties of immunoglobulin somatic mutation. PMID- 7486521 TI - Variable-region gene analysis of B-cell subsets derived from a 4-year-old child. Somatically mutated memory B cells accumulate in the peripheral blood already at young age. PMID- 7486522 TI - B-lineage differentiation stages resolved by multiparameter flow cytometry. PMID- 7486524 TI - Pattern of usage of the VH4-21 gene by B lymphocytes in a patient with EBV infection indicates ongoing mutation and class switching. PMID- 7486523 TI - Analysis of hypermutation in a mixed T-cell receptor beta/immunoglobulin heavy chain passenger transgene. PMID- 7486525 TI - Development of the antibody repertoire in rabbits. PMID- 7486526 TI - Development of the mouse B-cell repertoire. AB - The mouse B-cell repertoire early in ontogeny contains B cells with receptor immunoglobulins exhibiting high connectivity, multi/self-reactivity, and generally low affinity. This is due structurally to extensive restriction in the germline components used to generate the B-cell receptor. The selective pressure acting on the nascent repetoire has both negative and positive components as seen in our in vivo models. VH81X-bearing B cells from the VH81X transgenic mice (and probably from normal mice) are subject to self-selective pressure with two components: a positive one favoring a certain (self-reactive) specificity in the CD23-IgM+ population and a negative one preventing the entry of B cells with this specificity into the CD23+IgM+ compartment. PMID- 7486527 TI - Analysis of immunoglobulin gene rearrangement in fetal B-cell progenitors developing in vivo or in vitro. PMID- 7486529 TI - Analysis of DHQ52 gene segment transcription and rearrangement during B-cell development in human fetal bone marrow. PMID- 7486528 TI - Development of the antibody repertoire as revealed by single-cell PCR of FACS sorted B-cell subsets. PMID- 7486530 TI - Fetal versus adult PreB or B cells: the human VH repertoire. AB - At the preB stage, when only the IGH locus has rearranged, mu chains become expressed in association with the psi L chains, lambda-like and VpreB, thus forming the preB receptor. By the use of a monoclonal anti VpreB antibody, preB cells were isolated from two adult bone marrow samples, and the VH repertoire was analyzed and compared to fetal, XLA (X-linked agammaglobulinemia), and adult B repertoires. Most VH genes identified were also expressed in fetal liver, XLA bone marrow, and adult PBLs, with similar predominant usage of certain germline genes. Multiple D/D fusions, limited N diversity, and preferential use of JH4 with a low level of DQ52 usage were also identified. Few mutations could be observed, not specifically localized in CDR regions, that could be interpreted as not positively selected. Conversely, a shorter length of CDR3 appeared to be the hallmark of the preB step. Thus, the association of psi L chains with mu does not bring about a bias in the VH gene usage, but a first selection on the CDR3 region could be the result of recognition by given autoantigens or ligands different for preB cells and B cells. PMID- 7486531 TI - Developmental regulation of the human antibody repertoire. AB - The ability to respond to antigen develops in a programmed fashion during ontogeny. In human, "fetal" immunoglobulin gene segment utilization appears biased towards a small set of evolutionarily conserved V gene segments. Many of these gene segments are also used in antibodies with antigen specificities that do not arise until after infancy. The human fetus primarily regulates the diversity of the antibody repertoire through control of the H (heavy) chain CDR 3, which is generated by VDJ joining and forms the center of the antigen-binding site. Molecular modeling suggests that limitations in the length and composition of fetal CDR 3 intervals result in antibodies that contain a relatively "flat" antigen-binding surface that could serve to maximize the number of different interactions possible between the antibody and potential antigens. We propose that these limitations in the sequence and structure of H chain CDR 3 contribute to the low affinity and multireactivity of fetal antibody repertoires. The specific mechanisms used to generate a restricted fetal repertoire appear to differ between human and mouse. Nevertheless, included in the final products of both human and mouse fetal B cells will be antibodies that are quite homologous in composition and structure. The precise role that these antibodies play in the development of immunocompetence remains to be elucidated. PMID- 7486532 TI - Identification of genes specifically expressed at discrete stages of B-cell development. PMID- 7486533 TI - Human IgH-V gene repertoire in neonatal cord blood, adult peripheral blood, and EBV-transformed cells. PMID- 7486534 TI - The expressed heavy chain V gene repertoire of circulating B cells in normal adults. PMID- 7486536 TI - Molecular analysis of VH regions expressed in the adult B-cell repertoire. PMID- 7486537 TI - VH4.21 expression in the normal human B-cell repertoire. PMID- 7486538 TI - A complete preimmune human VH3 repertoire. PMID- 7486535 TI - The role of Bruton's tyrosine kinase in B-cell development and function in mice and man. PMID- 7486539 TI - Contribution of the V lambda light chain to the development of the primary antibody repertoire. AB - The experiments outlined here provide evidence that most VH gene families are expressed in association with either of three V lambda genes available in kappa deficient mice, though their expression is nonrandom. An exclusive use of V lambda L-chain genes, where predominantly expressed V lambda 1 and V lambda 2 differ by only seven amino acid substitutions in the germline, results in a higher degree of self-reactive antibodies to conserved natural antigens like albumin, thyroglobulin, and stress proteins. The higher levels of self-reactive antibodies, characteristic of preimmune B-cell repertoire, are likely to be a product of nonrandom VH gene family expression and their restricted pairing with three V lambda L-chain genes. These data suggest that a low lambda L-chain expression in normal mice is due, in part, to endogenous selection against lambda L chain in order to restrict self reactivity within the homeostatic threshold. PMID- 7486540 TI - Analysis of V kappa I and V lambda II light chain genes in the expressed B-cell repertoire. AB - Our previous studies of anti-DNA antibodies in SLE have demonstrated a preferential use of V kappa I and V lambda II gene families to encode light chains of antibodies that express the anti-DNA-associated 3I and 8.12 idiotypes, respectively. In this study, we employed PCR to obtain V kappa I and V lambda II germline genes from lupus patients in order to compare the germline genes to genes encoding expressed V kappa I and V lambda II light chains and to analyze the extent of somatic mutation among autoantibodies that derive from these light chain families. Our analysis shows that the germline repertoire among all persons (autoimmune and healthy) is comparable and that somatic mutation is used to diversify autoantibodies as well as anti-microbial antibodies. We have observed that autoantibodies encoded by V kappa I and V lambda II genes have a higher number of amino acid replacements in CDRs than autoantibodies encoded by other VL gene families. In addition, there may be subtle differences in V gene usage that distinguish the V kappa I-encoded light chains from other expressed V kappa light chains. PMID- 7486541 TI - Anomalous diversification of the antibody repertoire following bone marrow transplantation. PMID- 7486542 TI - Structure and thermodynamics of antigen recognition by antibodies. PMID- 7486545 TI - Effect of a putative B cell superantigen on complement. PMID- 7486546 TI - Structural correspondences between mouse and human immunoglobulin VH genes. Application to the humanization of mouse monoclonal antibodies. PMID- 7486547 TI - Structure of the VH-D-JH segments of human natural polyreactive IgM and IgG antibodies. Use of germline or somatically mutated forms of commonly expressed VH genes. PMID- 7486544 TI - Unconventional B-cell antigens and human immune repertoires. PMID- 7486549 TI - The immunoglobulin gene repertoire to Haemophilus influenzae type b. PMID- 7486543 TI - Analysis of the structural correlates for self-antigen binding by natural and disease-related autoantibodies. In vitro expression of recombinant and/or mutagenized human IgG. PMID- 7486548 TI - Structure-function studies of human monoclonal antibodies to pneumococcus type 3 polysaccharide. PMID- 7486550 TI - Patterns of V-region gene segment association in the human antibody response to Haemophilus influenzae type B capsular polysaccharide. PMID- 7486552 TI - Origins of anti-DNA antibodies and their implications for B-cell tolerance. PMID- 7486551 TI - Generation of diversity in a human anti-viral response. PMID- 7486553 TI - Ig mu heavy chains with VH81X variable regions do not associate with lambda 5. PMID- 7486554 TI - Does immunological tolerance explain the waste in the B-lymphocyte immune system? Experiment and theory. PMID- 7486555 TI - B-1 cells and autoimmunity. PMID- 7486556 TI - B-1 cellular origin and VH segment structure of IgG, IgA, and IgM anti-DNA autoantibodies in patients with systemic lupus erythematosus. PMID- 7486557 TI - The importance of the diversity region YYGS motif in anti-DNA antibody binding to DNA. PMID- 7486559 TI - A complete map of the human immunoglobulin VH locus. PMID- 7486560 TI - Immunoglobulin gene use by naturally occurring cold agglutinins. PMID- 7486558 TI - Dual recognition of lipid A and DNA by human antibodies encoded by the VH4-21 gene. A possible link between infection and lupus. PMID- 7486561 TI - Human pathogenic autoantibodies and their escape from deletion. PMID- 7486562 TI - V-gene repertoire and hypermutation of rheumatoid factors produced in rheumatoid synovial inflammation and immunized healthy donors. AB - We have compared RF from normal, immunized donors and RF derived from the synovial tissues of RA patients. We have found a difference in the preferential use of VL and VH gene families. In both conditions, RFs were found to have accumulated somatic mutations. However, there was a striking difference in the patterns of mutation. RFs from normals were characterized by a very low R:S ratio in the CDR1+2, considerably lower than seen among the RARFs. In addition, there was little increase in affinity with increasing numbers of mutations in a group of clonally related RFs from an immunized normal. This contrasts with RF from RA, where there is evidence of both affinity maturation and class switching. Together these data suggest that in healthy persons there is a controlling mechanism to limit the affinity of RF autoantibodies, and that this is lost in RA. The higher affinity of the RA-derived RF may be of significance in the pathology of the disease. PMID- 7486563 TI - Analysis of immunoglobulin gamma heavy chains from rheumatoid arthritis synovium. Evidence of antigen-driven selection. PMID- 7486564 TI - In situ hybridization analysis of immunoglobulin VH gene family expression in rheumatoid arthritis. PMID- 7486565 TI - Immunoglobulin gene usage in diabetes-associated human monoclonal antibodies recognizing glutamate decarboxylase. PMID- 7486566 TI - Expression and regulation of clonally ignorant B cells. PMID- 7486567 TI - Analysis of immunoglobulin VH gene repertoire by an anchored PCR-ELISA. AB - We developed a novel technique to analyze the relative concentration of the expressed immunoglobulin (Ig) VH genes using an enzyme-linked immunosorbent assay (ELISA). Expressed Ig cDNA are amplified via anchored PCR and then subjected to a "nested PCR" reaction that attaches biotin to the 5' end of the antisense strand. This allows us to tether the antisense strand of PCR products onto avidin-coated ELISA plates. Digoxigenin-labeled oligonucleotides specific for the leader sequence sense strand of each major Ig VH gene subgroup are used to probe the plate-tethered, alkaline-denatured, and single-stranded antisense cDNA. Bound probes then are detected with alkaline-phosphatase-conjugated anti-digoxigenin antibodies. Using this method, we assessed the distribution of Ig VH genes used by IgM-expressing blood B cells of normal adults. We found the predominant subgroup is VH3, representing approximately half (range 41-59%) of the expressed IgM repertoire. The next largest subgroups used are VH4 (19-23%), VH1 (15-17%), and VH5 (7-11%). The VH2, VH6, and VH7 subgroups each constitute less than 3% of the expressed IgM repertoire. These results agree with those obtained using traditional and more laborious methods that analyze the distribution of Ig clones in cDNA libraries. In addition, we find that this method compares favorably in sensitivity and specificity to more conventional techniques for assessing the clonality of blood or tissue B-cell populations. This rapid and nonradioactive method should have utility for assessing the Ig repertoires expressed by normal, autoimmune, or neoplastic B-cell populations. PMID- 7486568 TI - LIGM-DB/IMGT: an integrated database of Ig and TcR, part of the immunogenetics database. PMID- 7486569 TI - Expression of immunoglobulin heavy chain variable genes in B-chronic lymphocytic leukemia. PMID- 7486570 TI - In vivo isotype class switching in CD5+ chronic lymphocytic leukemia B cells. PMID- 7486571 TI - Clonal heterogeneity in membrane expression of CD5 in a patient with chronic lymphocytic leukemia. PMID- 7486572 TI - Clonal analysis of IgM+ CD5+ CLL B cells. PMID- 7486573 TI - Surface phenotype and immunoglobulin heavy chain gene usage in chronic B-cell leukemias. PMID- 7486574 TI - Different clonal origin of B-cell populations of chronic lymphocytic leukemia and large-cell lymphoma in Richter's syndrome. AB - Richter's syndrome is defined as the morphologic transformation of chronic lymphocytic leukemia (CLL) into diffuse large-cell lymphoma (DLL). To determine the clonal nature of the two malignancies, we microdissected the CLL and DLL cells from a lymph node of Richter's syndrome and analyzed the sequences of the rearranged Ig VH-D-JH genes of the two lymphomas. Using the Ig VH-D-JH sequence as a marker of clonality, we delineated the clonal relationship of the CLL and DLL cells. The microdissected CLL and DLL cells productively rearranged different VH, D, and JH genes, suggesting that these DLL B cells emerge as discrete elements independent of the CLL B-cell population. The productively rearranged Ig V gene sequence of the CLL clone was 100% identical to the VH6 germline gene, but the rearranged Ig VH gene of the DLL clone was somatically point-mutated based on comparison of its sequence with those of reported germline genes. In the DLL clone, the random distribution and nature of the somatic point-mutations suggests a lack of antigen selection; the identity of the somatic point-mutations in multiple independent isolates of the same B-cell clone suggests a lack of intraclonal diversity. Thus, Richter's syndrome DLL B cells are monoclonal and can emerge as discrete elements independent of the preexisting CLL cells; antigen selection and clonal diversification are not necessarily associated with the events leading to this aggressive neoplastic transformation. PMID- 7486575 TI - Polymorphism and utilization of human VH Genes. AB - The human VH germline repertoire comprises approximately 100 elements, which can be grouped into seven families based on nucleotide sequence similarity. Members of different families are interspersed throughout the complex, with limited sets of alleles identified for most loci. Linkage disequilibrium between most elements is weak. Variation within the population can be attributed to differences in nucleotide sequence between allelic genes as well as to differences in the number of genes present. Gene number per haplotype varies as a result of the common occurrence of insertion/deletion polymorphisms, which may be small, involving a single element, or may be extensive, involving four or five elements. In some cases, such polymorphisms may involve duplication of a functional VH gene segment on some haplotypes and deletion of the gene on others. The resulting variation in germline composition of the VH locus may have profound effects on VH gene utilization. PMID- 7486576 TI - VH gene repertoire of mantle cell lymphomas. PMID- 7486577 TI - VH gene use by CD5+ AIDS-associated B-cell lymphoproliferations. PMID- 7486579 TI - Analysis of multiple myeloma third complementarity-determining regions reveals characteristics of prenatal B cells. PMID- 7486578 TI - Cellular origin, antigen reactivity, and VH segment structure of IgM mAbs from AIDS lymphomas. AB - In the present studies we analyzed the Ag specificity, VH gene structure, and cellular origin of three IgM mAb-producing cell lines established in vitro from bioptic specimens of three AIDS patients with BL. We found that (i) both HBL-2 and HBL-3 IgM mAbs were cold agglutinins highly specific for the i blood group determinant, a self Ag the expression of which is dominant in the early stages of life, and both mAbs used somatically point-mutated VH 4-21 segments; (ii) HBL-1 IgM mAb, the Ag-specificity of which has not been determined, used a putatively mutated member of the VHIII family; and (iii) both HBL-1 and HBL-2, but not HBL 3, cells expressed CD5 mRNA, suggesting a B-1 cell origin. The utilization of VH4 21 by the HBL-2 and HBL-3 cold agglutinins is consistent with the usage of this gene segment by all the reported pathogenic except the naturally occurring cold agglutinins. This restricted VH gene usage may reflect an inherent affinity of the germline VH4-21 gene product for the i/I carbohydrate structure, and, perhaps, an overrepresentation of VH4-21 in the human early and late B-cell repertoire. Consistent with both an early and late developmental expression of the VH4-21 gene is the B-1 and B-2 cellular origin of the two VH4-21+ cold agglutinins reported here. Thus, the two cold agglutinin autoantibodies possibly emerged at different stages of the natural history of the B-cell repertoires of these patients and might display a different temporal relationship, as discussed below, to the time of emergence of the respective tumoral cells. The somatically mutated status of the HBL-2 and HBL-3 mAb VH segments was suggested by the monomorphism of the human VH4-21 gene, the extension of the nucleotide differences to the, in general, highly conserved JH segment; and it was formally proved in HBL-3 mAb. Positive selection by Ag of the R mutations in the HBL-2 and HBL-3 mAb VH segments was suggested by the differential R:S mutation ratios in the CDRs and FRs (HBL-2 mAb, 5.0 and 1.1, respectively; HBL-3 mAb, 2.2 and 0.3, respectively) but not substantiated by appropriate statistical analysis according to the binomial distribution model.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 7486580 TI - Hodgkin's disease: clonal Ig gene rearrangements in Hodgkin and Reed-Sternberg cells picked from histological sections. PMID- 7486581 TI - Production of antigen-specific human antibodies from mice engineered with human heavy and light chain YACs. AB - Our paper describes the introduction of large fragments of both the human heavy and light chain Ig genes into the mouse germline to create a mouse strain capable of producing a broad repertoire of antigen-specific, fully human antibodies. The human immunoglobulin gene sequences were functional in the context of the mouse machinery for antibody recombination and expression, either in the presence or absence of functional endogenous genes. This was demonstrated by their ability to undergo diverse rearrangement, to be expressed at significant levels, and to exclude expression of mouse immunoglobulins irrespective of their copy number or site of integration. The decrease in susceptibility to influence by adjacent genomic sequences may reflect the greater size, variable gene content, or structural integrity of the human Ig YACs and/or the presence of unidentified but important regulatory elements needed for optimal expression of the human immunoglobulin genes and their correct regulation. Our results show that mouse B cells coexpressing human heavy and kappa chains, upon immunization, can produce antigen-specific, fully human antibodies. Furthermore, the human heavy and kappa chain YACs induced differentiation and maturation of the growth-arrested B-cell lineage in mice with inactivated endogenous Ig genes, leading to the production of a diverse repertoire of fully human antibodies at levels approaching those in normal serum. These results suggest the potential value of these mice as a source of fully human antibodies for human therapy. Furthermore, it is expected that such mice would lack immunological tolerance to and thus readily yield antibodies to human proteins, which may constitute an important class of targets for monoclonal antibody therapy. Our findings suggest that the introduction of even larger portions of the human heavy and light chain loci, which should be achievable with the ES cell-yeast spheroplast fusion technology described, will result in strains of mice ultimately capable of recapitulating the full antibody repertoire characteristic of the human humoral response to infection and immunization. The present and future mouse strains may prove to be valuable tools for studying the molecular mechanisms and regulatory sequences influencing the programmed assembly and expression of human antibodies in the normal immune response, as well as the abnormal response characteristic of autoimmune disease and other disorders. The strategy we have described for the introduction of large segments of the human genome into mice in conjunction with the inactivation of the corresponding mouse loci may also have broad applicability to the investigation of other complex or uncharacterized loci. PMID- 7486582 TI - Class switching in human immunoglobulin transgenic mice. AB - We have introduced human germline-configuration heavy and kappa light chain minilocus transgenes into mice that have been engineered so that their endogenous heavy and kappa light chain loci are inactive. The two human transgenes are inserted by pronuclear microinjection, while the two endogenous mouse genes are disrupted by homologous recombination in embryonic stem cells. The resulting animals contain four unlinked genetic modifications and must rely on the introduced transgenes for the development of the B-cell lineage and for the generation of an antibody repertoire. The heavy chain transgene includes both the human mu and the human gamma 1 constant region gene segments, as well as upstream switch region sequences. Although mouse B cells and human B cells exhibit species specific differences in the induction of gamma isotype expression, the transgenic mouse B cells appear to undergo regulated switching to human gamma 1 both in vivo and in vitro. This observation defines a subset of the heavy chain constant region that is sufficient for class switching, and implies that the human gamma 1 switch region includes a core of sequence that is functionally homologous to those cis-acting regulatory elements that direct mouse gamma switching. PMID- 7486584 TI - Genetic and functional characterization of human autoantibodies using combinatorial phage display libraries. PMID- 7486583 TI - Isolation of human anti-red blood cell antibodies by repertoire cloning. PMID- 7486586 TI - Substrate-driven formation of a proteolytic antibody light chain. PMID- 7486585 TI - Repertoire cloning of human lupus autoantibodies. PMID- 7486587 TI - Catalytic activity of anti-thyroglobulin antibodies. PMID- 7486588 TI - Localization of protease activity in antibody subunits. PMID- 7486589 TI - Use of a phage expression system to produce Fabs from human autoanti-DNA antibodies encoded by the VH4-21 gene segment. PMID- 7486590 TI - The development of a bacteriophage expression vector for cloning immunoglobulin variable region genes from B-cell genomic DNA. PMID- 7486591 TI - Characterization and genomic mapping of a novel leader peptide associated with the human VH4-21 (VH4-34) gene segment. AB - The human IgVH locus is located on chromosome 14, band q32 and spans approximately 1 mb. Within this locus are approximately 120 VH gene segments that are subdivided into six to seven families based on sequence homology of their coding regions. VH4-21 (VH4-34) is a member of the VH4 family, a family that contains 10 to 15 members. It is expressed in a variety of circumstances including early fetal development, the autoantibody repertoire, and in a highly restricted manner in antibodies that recognize alloantigens on the surface of human red blood cells. Most interesting, however, is the expression of this gene segment in T cells as a semi-germline transcript in conjunction with a nontraditional VH leader peptide. This nonhydrophobic leader sequence, termed "Et" for exon in T cells, has previously been shown to reside within the VH locus. Using YAC and P1 clones, we have identified two copies of this exon, both of which are located in the region of the locus that contains VH4-24 (VH4-34). Characterization of the two exons suggests that they arose by duplication, as flanking DNA is almost identical over a distance of > 5 kb. Preliminary data suggests they are both located > 20 kb upstream of VH4-21 (VH4-34). PMID- 7486592 TI - Ethnic differences in VH gene polymorphism. PMID- 7486593 TI - Expression and hypermutation of human immunoglobulin kappa genes. PMID- 7486594 TI - Regulation of human lambda light chain gene expression. PMID- 7486595 TI - In vitro studies of human B lymphopoiesis. PMID- 7486596 TI - V(D)J recombination: double-strand break repair gene products used in the joining mechanism. PMID- 7486597 TI - Clinical potential for the use of neuroprotective agents. A brief overview. AB - "Stroke treatment seems to be entering a golden age ...." Fisher's observation not only applies to ischemic stroke, but to all the conditions described above, and in the future, possibly (and quite speculatively), to other neurologic diseases, such as multiple sclerosis, amyotrophic lateral sclerosis, even radiation therapy and Bell's palsy. Physicians must sharpen their criteria for decisions regarding therapy and must" ... be prepared to accept what is actually known from scientific data ... rather than to rely on instinct, clinical impression, or the need to do something rather than nothing." PMID- 7486598 TI - Neuroprotective effects of free radical scavengers and energy repletion in animal models of neurodegenerative disease. PMID- 7486599 TI - Neuroprotective effects of radical scavengers in an intact dorsal root ganglion hypoxia model. PMID- 7486601 TI - Potential interactions between nimodipine and adrenal hormones. PMID- 7486600 TI - Short-term regulation of neuronal calcium channels by depolarization. AB - The 1,4-dihydropyridine-sensitive voltage-gated Ca2+ channel is widely distributed in excitable cells. The channel and its several associated drug binding sites are known to be up- and downregulated by a variety of homologous and heterologous influences including membrane depolarization. The neurosecretory GH4C1 cell line possesses L-type channels. Depolarization of these cells by elevated K+ increases the binding affinity of 1,4-dihydropyridines and decreases the number of 1,4-dihydropyridine binding sites and functional channels. There is a coordinate upregulation of the number of muscarinic receptors. This membrane potential- and Ca(2+)-calmodulin-dependent process of channel downregulation may involve internalization of the channel heteromeric complex or, more plausibly, a dissociation of the complex and a concomitant loss of both binding and permeation functions. PMID- 7486602 TI - The effects of nimodipine on the EEG of substance abusers. AB - Cocaine abusers have increased EEG beta and areas of reduced cortical blood flow. Since, nimodipine has neuroprotective effects and increases blood flow, we investigated the efficacy of single and multiple doses of the nimodipine in normalizing the EEG of substance abusers. Fourteen subjects received single (0, 30, 60 mg) and eleven received multiple daily (up to 150 mg in 12 hours) doses of nimodipine to determine whether this drug would increase EEG alpha and decrease beta in substance abusers. The EEG was recorded from eight scalp locations (F3, C3, P3, O1, F4, C4, P4 and O2) for three minutes during eyes closed, and eyes open conditions. Single and multiple doses of nimodipine produced significant increases in EEG alpha and decreases in EEG beta in the eyes open condition. Thus, nimodipine may have potential therapeutic implications in the treatment of cocaine dependence. Chronic nimodipine dosing in cocaine-dependent individuals is now needed to confirm its efficacy in the treatment of cocaine dependence. PMID- 7486603 TI - Nimodipine improves information processing in substance abusers. AB - We examined whether nimodipine can improve information processing in healthy drug abusers using cognitive event-related potential (ERP) methodology. Placebo and 30 and 60-mg doses of nimodipine were administered on separate days in a random double-blind design to twelve male subjects, who used cocaine and/or opiates as well as alcohol and marijuana. The subjects performed the auditory rare event monitoring (AREM) task and the paired letter version of the visual continuous performance task (CPT) before oral drug administration as well as one and two hours after drug ingestion. The EEG was recorded from 7 scalp locations. The P3 component of the ERPs to the target stimulus was reduced with repeated testing on the placebo day. The 30-mg dose of nimodipine blocked the decrease in P3, which reflects stimulus evaluation in both tasks. Chronic administration of nimodipine may alleviate the cognitive deficits observed in substance abusers during abstinence and prevent treatment relapse. PMID- 7486605 TI - Dexamethasone and the prevention of neonatal hypoxic-ischemic brain damage. PMID- 7486607 TI - Prophylactic pharmacologic neuroprotection against focal cerebral ischemia. AB - The crucial importance of the earliest possible treatment of ischemic stroke has been stressed. This leads to the concept of prophylactic neuroprotection: long term administration of neuroprotective agents to people at risk for stroke, so that the effects of focal ischemia are countered from the moment of onset of vascular occlusion. We at NYU (and researchers at several other centers) have been testing possible neuroprotective agents in the cardiopulmonary bypass setting. We believe that by such means we can rapidly and economically screen agents and dose schedules for efficacy. We believe that an agent demonstrably reducing mental impairment after cardiopulmonary bypass suggests that testing such an agent in a prospective manner in people at risk for stroke would be productive, and we propose a sequence for such trials: people during and in the first month after carotid endarterectomy, then postcarotid endarterectomy patients during long-term follow-up, and finally populations at risk for spontaneous stroke in the nonvascular surgery setting. PMID- 7486606 TI - Risk assessment strategies for neuroprotective agents. AB - Neurotoxicity may be defined as any adverse effect on the structure or function of the central and/or peripheral nervous system by a biological, chemical, or physical agent. Neurotoxic effects may be permanent or reversible, produced by neuropharmacological or neurodegenerative properties of a neurotoxicant, or the result of direct or indirect actions on the nervous system. A multidisciplinary approach is necessary to assess neurotoxicity because of the complexity and diverse functions of the nervous system. Many of the relevant effects can be measured directly by neurochemical, neurophysiological, and neuropathological techniques, whereas, others must be inferred from observed behavior. Some neurotoxicological data can be derived directly from humans. Neurotoxicity in humans is most commonly measured by relatively noninvasive neurophysiologic and neurobehavioral methods that assess cognitive, affective, sensory, and motor function. For most toxicological assessments, however, it is necessary to rely on information derived from animal models. There are many approaches that can be used to assess neurotoxicity, including whole animal (in vivo) and tissue/cell culture (in vitro) testing. Neurotoxicity can be described at multiple levels of organization, including neurochemical, anatomical, physiological, and behavioral. An important aspect of neurotoxic endpoint evaluation involves risk assessment procedures. Risk assessment may be defined as an empirically-based process used to determine the probability that adverse or abnormal effects are associated with exposure to a chemical, physical or biological agent. Risk management, on the other hand, is the process that applies information obtained through the risk assessment process to determine whether the assessed risk should be reduced and, if so, to what extent. For chemicals such as neuroprotective agents and other drugs designed to provide therapeutic benefits, information concerning these benefits is considered during the risk management phase. The risk assessment process usually involves four steps: hazard identification, dose-response assessment, exposure assessment, and risk characterization. Neurotoxicity risk assessment models of the future may well include biomarkers of both effect and exposure as well as biologically-based mechanistic and pharmacokinetic considerations derived from both epidemiologic and experimental data. PMID- 7486604 TI - Adenosine: a prototherapeutic concept in neurodegeneration. PMID- 7486609 TI - Receptor subtypes linked to metabotropic glutamate receptor agonist-mediated limbic seizures in mice. PMID- 7486608 TI - Neuroprotective use-dependent blockers of Na+ and Ca2+ channels controlling presynaptic release of glutamate. AB - We have originated a family of N,N'-disubstituted guanidines that block the voltage-activated Ca2+ and Na+ channels governing glutamate release. These compounds, CNS 1237 (N-acenaphthyl-N'-methoxynaphthyl guanidine) and its analogues, are "use dependent" in their ability to attenaute neurotransmitter release: they block glutamate release with greater efficacy under conditions of persistent or repetitive depolarization, as would be encountered under pathophysiological circumstances, relative to their ability to block glutamate release elicited by brief, transient depolarizations more characteristic of normal physiological release events in nonischemic brain. Using electrophysiological and rapid kinetic methods, we have differentiated the use dependent block of the relevant Na+ and Ca2+ channels governing neurotransmitter release from the mechanism of channel antagonism exhibited by, respectively, the substituted guanidine Na+ channel blocker tetrodotoxin (TTX) and venom peptide Ca2+ antagonists. To characterize use-dependent Na+ channel block by CNS 1237, we have employed whole-cell voltage-clamp recordings from a Chinese hamster ovary (CHO) cell line expressing cloned mammalian type II Na+ channels. These experiments demonstrated that, in contrast to the actions of TTX under the same conditions, the potency of Na+ channel block by CNS 1237 is greatly enhanced by depolarizing stimuli in a frequency-dependent manner. Ca2+ channel-activated glutamate release from brain nerve terminal preparations was measured with approximately 300 msec time resolution over a 5-second period of high K(+) depolarization, using a rapid superfusion technique. CNS 1237 and analogues, at 1 3 microM, accelerated the decay of glutamate release by 40-70%, reflecting depolarization-induced enhancement of block. In contrast, blockade of glutamate release by the Ca2+ channel antagonist peptide toxins omega-aga IV-A (from spider venom) and omega-conotoxin M-VII-C (from cone snail venom) exhibited "reverse-use dependence:" at concentrations of 0.3 microM, which blocked the initial amplitude of glutamate release by 40-60%, the decay time constant for glutamate release was significantly increased, indicating depolarization-induced relief of block. These findings establish that CNS 1237 and other members of this compound series are use-dependent blockers of the voltage-activated ion channels governing glutamate release. Studies of CNS 1237 in the rat middle cerebral artery occlusion (MCAO) focal stroke model have indicated infarct size reduction comparable to that observed by the same investigators for the glutamate release blocker (BW 619C89 (Burroughs-Wellcome, now in clinical development). Maximal infarct size reduction is achieved with a 3-mg/kg bolus followed by a 4-hour infusion of 0.75 mg/kg/hr.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 7486610 TI - Neuroprotective properties of the uncompetitive NMDA receptor antagonist remacemide hydrochloride. PMID- 7486612 TI - The rationale for glutamate antagonists in the treatment of traumatic brain injury. AB - The recent development of potent antagonists for the most widespread neurotransmitter in the mammalian brain has opened up possibilities for many forms of therapy. The excitotoxic hypothesis implicates excessive release of excitatory amino acids (EAAs) as an important cause of brain damage, especially in acute ischemia, and chronic neurodegeneration. Focal ischemic damage and diffuse axonal injury are the major causes of brain damage after traumatic human brain injury. Evidence from animal models has shown that excitatory amino acid induced events maybe responsible for a proportion of the posttraumatic sequelae and that these effects can be blocked by EAA antagonists. This evidence is reviewed, and the implications for human pathophysiology and treatment are discussed. PMID- 7486611 TI - Safety, tolerability and pharmacokinetics of the N-methyl-D-aspartate antagonist Ro-01-6794/706 in patients with acute ischemic stroke. The Dextrorphan Study Group and Hoffmann-La Roche. AB - Dextrorphan HCl (Ro 01-6794/706) is an NMDA receptor antagonist with clinical potential for administration in an elderly population of acute ischemic stroke patients. In vivo experience with such patients demonstrated a consistent pharmacologic effect/adverse experience profile that is typical of an NMDA receptor antagonist (e.g., nystagmus, nausea, vomiting, agitation, somnolence, hallucinations and hypertension). For the most part, these pharmacologic effects were mild to moderate in severity; short-lived; reversible; not life-threatening and subjectively tolerated. The most serious pharmacologic effect produced by dextrorphan administration was hypotension, which occurred within a well-defined window of 90 minutes from the start of the loading dose infusion in patients who received 200 mg/hr or greater loading dose infusions. In all cases it was reversible without neurologic sequelae. Careful review of demographic and pharmacokinetic parameters did not demonstrate any overriding factor(s) to the production of hypotension other than the rate of the loading dose infusion. Severe hypotension, severe decreased levels of consciousness and respiratory depression should not be generally expected at loading doses less than 200 mg/hr. In summary, dextrorphan can be safely given to an elderly population of ischemic stroke patients as a loading dose rate below 200 mg/hr and as a maintenance dose rate between 50-90 mg/hr for 24 hours when patients are monitored carefully for pharmacologic effects. PMID- 7486613 TI - Strategies for neuroprotection with glutamate antagonists. Extrapolating from evidence taken from the first stroke and head injury studies. AB - Over fifty patients with severe head injury, and one hundred with stroke, have now been treated with the competitive NMDA antagonist CGS 19755 (selfotel). Preliminary analysis has shown possible evidence of benefit for both these clinical indications, and several other glutamate antagonists are now being evaluated for these indications in Phase II trials. The optimal dose of CGS 19755 (selfotel) for efficacy for severe head trauma has not yet been identified, but may be > 3 mg/kg, as at this dose there was evidence of an ICP lowering effect and improved CPP. For stroke, however, the maximal tolerated dose was 1.5 mg/kg, because these conscious patients developed hallucinations and agitation. There were no other significant drug-associated adverse events in either of these studies. It is difficult to determine the "neuroprotective" dose for this compound in humans. By extrapolating from animal studies the "best estimate" would be around 5 mg/kg in patients with severe head trauma. For stroke, behavioral side effects were the major limiting factor for dosing. Although several NMDA antagonists, including CGS 19755 (selfotel), are currently entering efficacy trials for stroke, based upon their tremendous potency in animal models, the problem of psychomimetic effects may necessitate the use of additional management strategies, e.g., more intensive monitoring and concomitant medications. PMID- 7486616 TI - Evidence for prolonged release of excitatory amino acids in severe human head trauma. Relationship to clinical events. PMID- 7486615 TI - A fetal rat model of acute perinatal ischemia-hypoxia. PMID- 7486614 TI - Clinical pharmacology of CNS 1102 in volunteers. AB - The high affinity noncompetitive N-methyl-D-aspartate receptor antagonist CNS 1102 (aptiganel hydrochloride, Cambridge NeuroScience, Cambridge, MA.) is neuroprotective in preclinical models of stroke when administered as pretreatment or up to 60 minutes postischemia, and has potential for treatment of acute stroke or traumatic brain injury in man. A total of 55 healthy male subjects have participated in three separate studies to determine the clinical pharmacology of CNS 1102, 43 of whom have received CNS 1102 in doses of up to 100 micrograms/kg. Administration of CNS 1102 has been studied as a 15-minute intravenous infusion, as a 15-minute loading intravenous infusion followed by a 4-hour maintenance infusion, or as a fixed-dose intravenous bolus over 90 seconds. CNS 1102 in normal volunteers is well tolerated in total doses up to 32 micrograms/kg whether as a bolus injection, 15-minute infusion or 4-hour infusion. Central nervous system affects are evident within minutes of administration, implying rapid drug penetration. CNS 1102 has a large and variable volume of distribution (mean +/- standard deviation, 6.2 +/- 1.9 l/kg), variable clearance (115 +/- 77 l/h), and plasma half-life of approximately 4.5 hours. Adjustment of doses by subject weight does not improve variability of these parameters, and fixed doses may thus be administered. CNS 1102 causes dose-dependent elevation of blood pressure, accompanied by clinical evidence of vasoconstriction. Global cerebral blood flow is maintained, whilst middle cerebral artery flow velocity increases. Symptoms of light-headedness, disorientation and paresthesia progress through euphoria, disinhibition, and hallucinations to psychomotor retardation, paranoia and catatonia as total administered dose increases. PMID- 7486617 TI - The neuroprotective effect of calcitonin gene-related peptide following subarachnoid hemorrhage. European CGRP in Subarachnoid Haemorrhage Study Group. PMID- 7486618 TI - A small animal model of focal cerebral ischemia for studying neuroprotective agents. PMID- 7486619 TI - Quantitative histological evaluation of neurotoxic hippocampal damage. PMID- 7486620 TI - Perinatal brain injury. Pathophysiology and therapeutic intervention. PMID- 7486621 TI - The role of the growth factors IGF-1 and TGF beta 1 after hypoxic-ischemic brain injury. PMID- 7486622 TI - Oxidative brain damage in aged mice. Protection by caloric reduction. PMID- 7486623 TI - The role of hyperthermia in amphetamine's interactions with NMDA receptors, nitric oxide, and age to produce neurotoxicity. PMID- 7486624 TI - Lack of mitigation of methamphetamine-induced neurotoxicity by ganglioside GM1 or vitamin E. PMID- 7486625 TI - Structural-functional correlates of neuroprotection in the aging rabbit by a calcium channel blocker. Nimodipine reverses neocortical dendritic atrophy and improves memory retention. PMID- 7486626 TI - Neuroprotective activity of HU-211, a novel nonpsychotropic synthetic cannabinoid. PMID- 7486627 TI - A randomized, double-blind, placebo-controlled pilot trial of intravenous magnesium sulfate in acute stroke. PMID- 7486628 TI - Efficacy and tolerability of lifarizine in acute ischemic stroke. A pilot study. Lifarizine Study Group. PMID- 7486629 TI - Considerations in the design of preclinical safety evaluation programs for novel therapeutics used in neurologic diseases. PMID- 7486630 TI - Sensitization and desensitization of the NMDA receptor complex. Implications for therapy. PMID- 7486631 TI - Initial experience with remacemide hydrochloride in patients with acute ischemic stroke. PMID- 7486632 TI - The tolerability, pharmacokinetics and pharmacodynamics of increasing intravenous doses of 619C89, a novel compound for the acute treatment of stroke, in healthy volunteers. PMID- 7486633 TI - N-Methyl-D-aspartate receptor participation in Parkinson's disease, a neurodegenerative disorder. PMID- 7486635 TI - Disposition and pharmacokinetics of remacemide hydrochloride in male Sprague Dawley rats. PMID- 7486634 TI - A randomized, double-blind, placebo-controlled ascending dose tolerance study of 619C89 in acute stroke. PMID- 7486636 TI - The cerebral hemodynamic and metabolic effects of the noncompetitive NMDA antagonist CNS 1102 in humans with severe head injury. PMID- 7486637 TI - Effects of nimodipine and verapamil on cerebral blood flow and cerebrovascular reactivity in conscious rabbits. PMID- 7486638 TI - Clinical pharmacology of CNS 1102 in man. PMID- 7486639 TI - Low environmental temperatures or pharmacologic agents that produce hyperthermia decrease methamphetamine neurotoxicity in mice. PMID- 7486640 TI - Biologically based dose-response model for neurotoxicity risk assessment. PMID- 7486641 TI - Role of reactive oxygen species (ROS) in neuronal degeneration. Modulation by protooncogene expression. PMID- 7486642 TI - Phospholipase A2 regulation in neural function and injury. PMID- 7486644 TI - Quantitative histological evaluation of neuroprotective compounds. AB - The application of quantitative morphometric methods to neurotoxicology is a relatively recent endeavor, and appropriate techniques are still evolving. However, such methods are essential for subsequent use of neurohistological data in mathematical representations of the risk of exposure to neurotoxicants. It can be predicted that the same methods will also be of great utility in studies of the efficacy of neuroprotective drugs. When the neuropathological conditions to be prevented or reversed are best monitored by neurohistology, quantitative morphometry should be considered as the most direct means to demonstrate the efficacy of a neuroprotective agent. Initially, a decision to choose the most appropriate histological procedure must be made. The rationale for such decisions with regard to several common histochemical techniques was discussed. The appropriate stereological and statistical considerations to be addressed by the sampling strategy were also presented. It is anticipated that quantitative morphometric methods will play an increasingly important role in the evaluation of the efficacy and toxicity of neuroactive compounds. PMID- 7486643 TI - Evaluation of the novel neuroprotective agent BW619C89 in the middle cerebral artery occlusion model of focal ischemia in the spontaneously hypertensive rat and normotensive Fischer 344 rat. PMID- 7486645 TI - Brain injury and inflammation. A putative role of TNF alpha. PMID- 7486646 TI - Comparison of glutamine-enhanced glutamate release from slices and primary cultures of rat brain. AB - Increased extracellular glutamate has been associated with a wide range of effects including production of neurotoxicity. Glutamine has previously been shown to cause increased release of glutamate from a variety of preparations. Extracellular central nervous system (CNS) glutamine levels are known to increase with neurotoxin exposures, hepatic failure, renal failure, head trauma or stroke. However, the action of glutamine to enhance the release of glutamate under nondepolarizing conditions has not been well studied. Since glutamine-mediated increases in extracellular glutamate are potentially of significance in cellular damage as a result of CNS insult, further examination of this phenomenon is important. Striatal and hippocampal slices or virtually neuron-free primary striatal glial cultures were employed in studies to further elucidate the mechanism(s) of glutamine-enhanced glutamate release. Elevated extracellular glutamine caused increased glutamate release in all three preparations. In hippocampal and striatal slices elevated glutamine caused an enhancement of N methyl-D-aspartate (NMDA) receptor-mediated [3H]catecholamine release equivalent to that produced by high concentrations (up to 100 microM) of exogenous glutamate. In both striatal slices and primary cultures kynurenate increased glutamate release in the presence of 500 microM glutamine, while kainate either had no effect or decreased glutamate levels in the presence of glutamine. Since several presynaptic modulators of release did not affect the glutamate release produced by glutamine in slices, vesicular release of glutamate from nerve terminals was probably not involved in the effects of the exogenous glutamine. The similarities between striatal slices and primary striatal cultures indicate that enzymatic conversion of glutamine to glutamate within glia may be an important factor in the glutamine-mediated elevation of extracellular glutamate levels. PMID- 7486647 TI - Effect of hypocapnia on extracellular glutamate and glycine concentrations during the periischemic period in rabbit hippocampus. AB - Glutamate (GLU) is a neurotransmitter. Massive release of GLU and glycine (GLY) into the brain's extracellular space may be triggered by ischemia, and may result in acute neuronal lysis or delayed neuronal death. The aim of this study was to evaluate the possible relationship between hyperventilation and the level of GLU and GLY during brain ischemia. Rabbits were anesthetized with halothane and oxygen. Group 1 was allowed to hyperventilate (PaCO2 25-35 mmHg). PaCO2 was maintained throughout the study. Group 2 was a normal control group that maintained normocapnia. Two global cerebral ischemic episodes were produced. Microdialysate was collected during the periischemic and reperfusion periods from the dorsal hippocampus. GLU and GLY concentrations were determined using high performance liquid chromatography. In the control group, GLU and GLY were significantly elevated during each episode of ischemia; these levels returned to baseline within 10 minutes after reperfusion. In contrast, in the hyperventilation group GLU and GLY concentrations increased during ischemia, but they were not statistically significant. Two way ANOVA for the periischemic periods (t = 15,80; p = 0.06) revealed lower GLU values for the hyperventilated animals. A similar analysis for periischemic GLY concentrations revealed significantly lower values in the hyperventilated group (t = 10,15,75,80: p = 0.03) as compared to normal controls. We were able to demonstrate that hypocapnia during periischemic period lowered extracellular GLU and GLY concentrations. These results can explain a part of the protective action of hypocapnia during cerebral ischemia. PMID- 7486649 TI - Coreceptors and adapter proteins in T-cell signaling. PMID- 7486648 TI - Definition of signals for neuronal differentiation. PMID- 7486650 TI - Symmetry of the activation of cyclin-dependent kinases in mitogen and growth factor-stimulated T lymphocytes. AB - The entry of resting T cells into the G1 phase of the cell cycle after stimulation by mitogens is controlled by a series of biochemical events that are independent of growth factors. These events follow the initial signals stimulated through the engagement of the T-cell receptor and include activation of the cyclin-dependent kinases Cdk6, Cdk4, and Cdk2, as well as a transient phosphorylation of the retinoblastoma gene product (p110Rb) by one or several of these proteins. A progression signal such as that delivered by interleukin-2 then induces a second phase of Cdk6, Cdk4, and Cdk2 activation, along with sustained phosphorylation of p110Rb in the activated T cells. This second signal is required to carry the cells into the S phase and beyond. Quantitative and qualitative differences in the expression and activity of these proteins may be critical to maintain the delicate balance that is necessary to ensure the normal progression of T cells through the cell cycle. PMID- 7486651 TI - Molecular and genetic insights into T-cell antigen receptor signaling. PMID- 7486652 TI - Signal transduction mediated by the T-cell antigen receptor. PMID- 7486653 TI - Genetic dissection of the transducing subunits of the T-cell antigen receptor. PMID- 7486654 TI - Structure and catalytic properties of protein tyrosine phosphatases. PMID- 7486655 TI - The cyclosporin-sensitive transcription factor NFATp is expressed in several classes of cells in the immune system. PMID- 7486656 TI - Signal transduction by the B-cell antigen receptor. AB - The antigen receptor of B lymphocytes (BCR) plays important roles in recognition of foreign antigens and self-components to allow the immune system to make appropriate antibody responses. The BCR is a complex between membrane immunoglobulin and the Ig-alpha and Ig-beta heterodimer. Site-directed mutagenesis experiments have shown that the mu heavy chain transmembrane domain plays a key role in the association of mIgM with Ig-alpha/Ig-beta. In the absence of complex formation, mIgM is retained in the endoplasmic reticulum, and this function is also specified by the mu chain transmembrane domain. The ability of various mutant mIgM molecules to associate with Ig-alpha/Ig-beta correlates well with their ability to induce signal transduction reactions such as protein tyrosine phosphorylation and phosphoinositide breakdown. Thus, the signaling ability of the BCR appears to reside in the Ig-alpha/Ig-beta heterodimer. The cytoplasmic domains of Ig-alpha and Ig-beta each contain an ITAM sequence, which is defined by its limited homology with subunits of the T-cell antigen receptor and of Fc receptors. Moreover, chimeric proteins containing these ITAMs and surrounding sequences from the cytoplasmic domains of Ig-alpha or Ig-beta exhibit signaling function characteristics of the intact BCR. The Ig-alpha and Ig-beta chimeras are each capable of inducing all of the BCR signaling events tested and thus represent redundant functions. Cross-linking these chimeras leads to their phosphorylation and to binding of the intracellular tyrosine kinases Lyn and Syk. The BCR expressed in the nonlymphoid AtT20 cells, which express the Src-family tyrosine kinase Fyn but not Syk, was not able to trigger vigorous signaling reactions. Introduction of the active form of Syk into these cells restored some signaling events. These results are consistent with a model in which the ITAMs act to initiate the BCR signaling reactions by binding and activating tyrosine kinases. PMID- 7486657 TI - Interaction of Shc with Grb2 regulates the Grb2 association with mSOS. PMID- 7486658 TI - A 115 kDa tyrosine phosphorylated protein associates with Grb-2 in activated Jurkat cells. PMID- 7486659 TI - Csk associates with the TCR zeta and epsilon chains through its SH2 domain. PMID- 7486661 TI - Potentiation of B-cell antigen receptor-mediated signal transduction by the heterologous src family protein tyrosine kinase, src. PMID- 7486662 TI - Regulation of human natural killer-cell lytic activity by serine/threonine phosphatases and kinases. PMID- 7486660 TI - Fc receptor stimulation of PI 3-kinase in NK cells is associated with protein kinase C-independent granule release and cell-mediated cytotoxicity. PMID- 7486663 TI - Mechanisms of enhanced nuclear translocation of the transcription factors c-Rel and NF-kappa B by CD28 costimulation in human T lymphocytes. PMID- 7486664 TI - Cytokine signal transduction through a homo- or heterodimer of gp130. PMID- 7486665 TI - The G protein-coupled receptor family and one of its members, the TSH receptor. PMID- 7486666 TI - IL-2 signaling involves recruitment and activation of multiple protein tyrosine kinases by the IL-2 receptor. AB - The IL-2 receptor (IL-2R) consists of three subunits, the IL-2R alpha, IL-2R beta, and IL-2R gamma chains, the last of which is also used in the receptors for IL-4, IL-7, IL-9, IL-13, and IL-15. The IL-2-induced proliferative signals emanate from the cytoplasmic domains of IL-2R beta and IL-2R gamma, but the nature and function of the signaling molecules that transmit these signals are not fully understood. Here we summarize our current understanding of the mechanisms by which IL-2R transmit signals by using multiple protein kinases. In fact, at least four protein tyrosine kinases (PTKs) are physically associated with IL-2R: p56lck (and its members), Syk PTK, and the Janus kinases, Jak1 and Jak3. cDNA expression studies revealed that the activation of these PTKs is critical for IL-2-induced proliferative signal transmission. Our findings indicate that a unique property of the IL-2R cytoplasmic domains is to recruit a variety of signaling molecules, which may suggest a mechanism by which these PTKs and other signaling molecules function in concert. PMID- 7486667 TI - Costimulation requirement for AP-1 and NF-kappa B transcription factor activation in T cells. AB - The transcriptional activity of the IL-2 promoter requires T-cell costimulation delivered by the TCR and the auxiliary receptor CD28. Several transcription factors participate in IL-2 promoter activation, among which are AP-1-like factors and NF-kappa B. Protein phosphorylation has an important role in the regulation of these two factors: (1) it induces the transactivating capacity of the AP-1 protein c-Jun; and (2) it is involved in the release of the cytoplasmic inhibitor, I kappa B, from NF-kappa B, allowing translocation of the latter into the nucleus. We have recently shown that both phosphorylation processes require T cell costimulation. Furthermore, in activated T cells, the kinetics of the two phosphorylation events are essentially similar. According to our results, however, the kinases responsible for the two processes are distinct entities. Whereas TPCK inhibits phosphorylation of I kappa B and, consequently, activation of NF-kappa B, it markedly enhances the activity of JNK, the MAP kinase-related kinase that phosphorylates the transactivation domain of c-Jun. We, therefore, propose the activation scheme presented in FIGURE 3 for T-cell costimulation. Costimulation results in the activation of a signaling pathway that leads to the simultaneous induction of the two transcription factors, AP-1 and NF-kappa B. Integration of the signals generated by TCR and CD28 engagement occurs along this pathway, which then bifurcates to induce I kappa B phosphorylation and NF-kappa B activation on the one hand, and JNK activation and c-Jun phosphorylation on the other. We are currently engaged in defining where the two signals integrate along the AP-1/NF-kappa B pathway. PMID- 7486668 TI - Structural aspects of cytokine/receptor interactions. PMID- 7486669 TI - Cell growth signal transduction is quantal. PMID- 7486670 TI - TNF alpha is an effective therapeutic target for rheumatoid arthritis. AB - Rheumatoid arthritis is the most common of a number of diseases in which inflammation and tissue destruction is driven by an autoimmune process. Current therapy is inadequate, and this has prompted major research efforts, both in academia and industry, to understand more about the pathogenesis, and hence provide the rationale for new therapeutic strategies. Here we review our studies of cytokine expression and regulation in rheumatoid joints, which has culminated in demonstrating that TNF alpha blockade, using a chimeric (human IgG1/K, mouse Fv) anti-TNF alpha antibody, cA2, markedly ameliorates arthritis. This defines a therapeutic target for rheumatoid arthritis. PMID- 7486671 TI - Investigation of ligand binding to members of the cytokine receptor family within a microbial system. PMID- 7486672 TI - The proline-rich motif is necessary but not sufficient for prolactin receptor signal transduction. PMID- 7486673 TI - Activation of multiple protein kinases by interleukin-1. PMID- 7486674 TI - IL-8 signal transduction in human neutrophils. PMID- 7486675 TI - IL-8 induces calcium mobilization in interleukin-2-activated natural killer cells independently of inositol 1,4,5 trisphosphate. PMID- 7486676 TI - Interleukin-11 induces tyrosine phosphorylation, and c-jun and c-fos mRNA expression in human K562 and U937 cells. PMID- 7486677 TI - Analysis of the interaction between two TGF-beta-binding proteins and three TGF beta isoforms using surface plasmon resonance. PMID- 7486678 TI - The stress-activated protein kinases. A novel ERK subfamily responsive to cellular stress and inflammatory cytokines. PMID- 7486679 TI - Inositol trisphosphate and calcium signaling. PMID- 7486681 TI - Structural aspects of receptor dimerization. c-kit as an example. PMID- 7486682 TI - Signal transduction interception as a novel approach to disease management. PMID- 7486680 TI - Protein serine/threonine kinases of the MAPK cascade. PMID- 7486684 TI - Mechanism of insulin and IGF-I receptor activation and signal transduction specificity. Receptor dimer cross-linking, bell-shaped curves, and sustained versus transient signaling. PMID- 7486683 TI - Common and distinct elements in insulin and PDGF signaling. AB - The receptors for insulin and PDGF are tyrosine kinases that mediate distinct effects in identical cellular backgrounds. Each receptor must therefore engage a unique subset of the available signaling elements--at least partly through the selection of proteins with src-homology 2 domains (SH2 proteins). Autophosphorylation sites in the PDGFr directly bind SH2 proteins, whereas activation of the insulin receptor leads to phosphorylation of IRS-1, which in turn binds SH2 proteins. In HIR 3.5 cells, which contain similar numbers of PDGF and insulin receptors, insulin, but not PDGF, stimulated tyrosyl phosphorylation of IRS-1. Similarly, insulin, but not PDGF, treatment of HIR 3.5 stimulated the association of IRS-1 with PtdIns 3'-kinase, although PDGF stimulated the association of PtdIns 3'-kinase with the tyrosine-phosphorylated PDGFr. Association with IRS-1 activated PtdIns 3'-kinase more effectively than association with the PDGFr. Whereas the PDGFr associated with PtdIns 3'-kinase, ras-GAP, GRB-2, and phospholipase C gamma, only GRB-2 and PtdIns 3'-kinase associated with IRS-1. Moreover, PDGF, but not insulin, caused tyrosine phosphorylation of phospholipase C gamma in HIR 3.5 cells. Thus, the insulin signal differs from that of PDGF by the insertion of a cytosolic, nonreceptor SH2 domain docking protein (IRS-1). Furthermore, IRS-1 binds a different subset of SH2 domain-containing proteins than does the PDGFr and regulates at least one common element (PtdIns 3'-kinase) differently than the PDGFr. These results support the hypothesis that IRS-1 differentiates the signals generated by the insulin receptor and PDGFr tyrosine kinases by binding and regulating a specific subset of SH2 domain-containing signaling molecules. PMID- 7486685 TI - The role of the insulin-like growth factor-I receptor in cancer. PMID- 7486686 TI - Mitogenic potential of insulin on lymphoma cells lacking IGF-1 receptor. AB - We have characterized an insulin-dependent T-cell lymphoma, LB, devoid of IGF-I receptor, which undergoes insulin stimulation and cell proliferation both in vitro and in vivo. In these cells, the mitogenic response can be evoked only through binding of insulin to its own receptor. This lymphoma is thus a good model for studying the molecular mechanisms involved in insulin mitogenicity. The high level of activated Ras in LB cells, even under nonproliferative conditions, shows that activation of Ras is insufficient for mitogenicity. It has been suggested earlier that separate pathways of signal transduction may emerge from Ras. The decision to activate a certain signaling pathway may depend on the activation state of other signaling routes in the cell. This may be the case in LB cells, where a signaling component activated by insulin works in concert with the Ras signaling pathway to induce mitogenesis. Yet it is still unclear whether activated Ras is a prerequisite for the insulin-induced response in LB cells. PMID- 7486687 TI - Platelet-derived growth factor. Distinct signal transduction pathways associated with migration versus proliferation. AB - Figure 2 summarizes our current interpretation of data concerning signals from the activated PDGF receptor involved in directed migration and proliferation of human arterial SMC. Binding of PDGF (PDGF-BB or PDGF-AA) causes PDGF-receptor dimerization, tyrosine autophosphorylation, and subsequent binding of several molecules containing SH2 domains to the activated receptor. Binding and activation of PLC gamma by the PDGF receptor leads to PIP2 hydrolysis, resulting in generation of diacylglycerol (DAG) and IP3. Subsequently, intracellular levels of calcium are elevated as a result of IP3-mediated calcium release from intracellular compartments. The decreased levels of PIP2 and increased levels of calcium both favor actin-filament disassembly by inducing capping of actin filament barbed ends and actin-monomer sequestration. A localized, and transient, actin-filament disassembly enables the cell to extend filopodia towards PDGF, thereby enabling chemotaxis to take place. At a later time and/or in a different compartment, actin-filament assembly is promoted by PDGF by a mechanism that is not completely understood, but that may involve small GTP-binding proteins, such as Rho, and formation of DAG. Migration on collagen requires functional alpha 2 beta 1 integrins, which may either constitute a permissive state required for a cell to migrate, or which may be actively involved in intracellular signals leading to migration. PDGF-induced DNA synthesis and proliferation involves activation of Ras, MAP kinase kinase, and MAP kinase. Cross-talk between PKA signaling and tyrosine-kinase receptor signaling results in PKA inhibition of the MAP kinase cascade, probably at the level of Raf. Activation of PI 3-kinase, or a PI 3-kinase-like enzyme, is also likely to contribute to the mitogenic effects of PDGF in these cells (Bornfeldt, unpublished observation). What determines if a SMC will migrate and/or proliferate in response to PDGF? Results are starting to emerge that show regulation of expression of molecules involved in intracellular signaling with different phenotypic states of SMC. For example, expression of PLC gamma is very low in intact vascular wall (where SMC show a "contractile phenotype"), and induced when SMC are converted to a "synthetic phenotype" in culture. Proliferation and expression of MAP kinase, but not calcium signaling, appear to be regulated by the extracellular matrix, and the profile of integrin expression is different in SMC in culture compared to SMC in the vascular wall. Thus, the relation between expression of signaling molecules involved in migration and signaling molecules involved in proliferation, as well as cross talk between different signal-transduction pathways, may determine the net effect of PDGF. PMID- 7486688 TI - Thrombin and its receptor in growth control. PMID- 7486689 TI - Interaction of signaling and trafficking proteins with the carboxyterminus of the epidermal growth factor receptor. PMID- 7486690 TI - Structural and functional properties of the TRK family of neurotrophin receptors. AB - The Trk family of tyrosine-protein kinases, TrkA, TrkB, and TrkC, are the signaling receptors that mediate the biological properties of the NGF family of neurotrophins. This family of growth factors includes in addition to NGF, BDNF, NT-3, and NT-4. TrkA is the NGF receptor. TrkB serves as a receptor for both BDNF and NT-4, and TrkC is the primary receptor for NT-3. NT-3 is a somewhat promiscuous ligand that can also activate TrkA and TrkB receptors at high concentrations. The trkB and trkC genes also encode noncatalytic receptor isoforms of an, as yet, unknown function. In addition to the Trk receptors, the NGF family of neurotrophins also binds with low affinity to an unrelated molecule, designated p75, a member of the TNF-receptor superfamily. Recently, we have generated strains of mice lacking each of these tyrosine-kinase receptors by gene targeting in embryonic stem cells. Characterization of these mutant mice is providing relevant information regarding the critical role that these receptors play in the ontogeny of the mammalian nervous system. PMID- 7486691 TI - Protein kinase C mediates short- and long-term effects on receptor tyrosine kinases. Regulation of tyrosine phosphorylation and degradation. PMID- 7486692 TI - Flag-insulin receptor mutants. PMID- 7486694 TI - The acute insulin-like effects of growth hormone in primary adipocyte-signaling mechanisms. PMID- 7486693 TI - Localization of specific amino acids contributing to insulin specificity of the insulin receptor. PMID- 7486695 TI - Regulatory interaction between calmodulin and the epidermal growth factor receptor. PMID- 7486696 TI - Phosphorylation of connexin-32 by the epidermal growth factor receptor tyrosine kinase. PMID- 7486697 TI - Molecular dissection of the growth hormone receptor. Identification of distinct cytoplasmic domains corresponding to different signaling pathways. PMID- 7486698 TI - Endothelin stimulates MAP kinase activity and protein synthesis in isolated adult feline cardiac myocytes. PMID- 7486699 TI - Control of T-cell development by the pre-T and alpha beta T-cell receptor. PMID- 7486700 TI - T-cell recognition of antigen. A process controlled by transient intermolecular interactions. AB - As recently as ten years ago, the nature of the T-cell receptor for antigen was a mystery, as was the precise role of histocompatibility molecules in antigen presentation to T cells. Although T-cell receptors have now been cloned and crystal structures of MHC/peptide molecules exist, our understanding of the parameters that characterize this interaction and other interactions relevant to T-cell immunity are still unclear. The engineering of soluble forms of proteins that mediate T-cell recognition of antigen has allowed the first measurements of these parameters. Interestingly, many of these interactions are of a transient nature, with very rapid off-rates. These data suggest a model whereby highly reversible intermolecular interactions mediate the cell-cell association. The association of adhesion molecules is probably the first step in the stabilization of a conjugate, because they are more numerous than any antigen-specific interaction, followed later by TCR-MHC engagements. Diffusion within each lipid bilayer should allow the congregation of MHC/TCR interactions at the cell-cell interface, with peptide-specific TCR interactions outcompeting irrelevant interactions. Rapid off-rates for both the antigen-specific and nonspecific interactions may be necessary to maintain reversibility, yet allow a rapid approach to equilibrium and consequent signaling when a specific antigen is present or disengagement when it is not. PMID- 7486701 TI - Receptors that regulate T-cell susceptibility to apoptotic cell death. AB - The immune system provides a useful model system in which to study the signal transduction events involved in the regulation of programmed cell death. Mature lymphocytes have the capacity to survive in the body for prolonged periods of time. During an immune response, cells of the appropriate antigenic specificity must undergo clonal amplification to mount a protective response, and cells participating in inflammatory immune responses need to have the capacity to survive at sites of inflammation. However, upon completion of a successful inflammatory response, the majority of cells produced must die off in order to maintain the homeostasis of the organism. Over the last several years we have learned a great deal about how mature lymphocytes regulated their susceptibility to undergo programmed cell death. Three types of information appear to be used by the lymphocyte to control its susceptibility to undergo programmed cell death. The intrinsic susceptibility of a cell to undergo programmed cell death is determined by members of the Bcl-2 gene family. In addition, extrinsic survival factors, such as IL-2, can initiate signal transduction events that can prevent a cell from initiating apoptosis. Finally, lymphocytes appear to have specific receptors, such as Fas, that can directly induce programmed cell death upon ligand binding. The integration of these three systems is discussed. PMID- 7486703 TI - Interaction of p56lck with CD4 in the yeast two-hybrid system. PMID- 7486702 TI - Signaling difference between class IgM and IgD antigen receptors. AB - Most mature B lymphocytes coexpress two classes of antigen receptor, IgM and IgD. The differences in the signal transduction from the two receptors are still a matter of controversy. We have analyzed B-cell lines expressing IgM or IgD antigen receptors with the same antigen specificity. Cross-linking of these receptors with either antigen or class-specific antibodies results in the activation of protein tyrosine kinases and the phosphorylation of the same substrate proteins. The kinetic and intensity of phosphorylation, however, was quite different between the two receptors when they were cross-linked by antigen. In membrane IgM-expressing cells, the substrate phosphorylation reached a maximum after one minute and diminished after 60 minutes, whereas in the membrane IgD expressing cells, the substrate phosphorylation increased further over time, reaching its maximum at 60 minutes and persisting longer than 240 minutes after exposure to antigen. Recently prolonged signaling has been found to be responsible for signaling differences between tyrosine kinase receptors using otherwise similar signaling routes. Thus, the duration of a signal may be an important biological feature of signal-transducing cascades. PMID- 7486704 TI - Induction of terminal differentiation of promyelocytic HL-60 leukemic cells implanted with lymphocyte receptors and stimulated with various lymphocyte stimulators. PMID- 7486705 TI - Independent signaling for growth arrest and apoptosis by Ig alpha and Ig beta subunits of the B-cell antigen receptor complex. PMID- 7486707 TI - [Audiometric results of 81 ossiculoplasties after tympanoplasty with closed technique in chronic cholesteatomatous otitis]. AB - We report our functional results about 81 ossiculoplastys during tympanoplasty with closed technique in chronic cholesteatomatous otitis. We discuss our functional results and the post-operative assessment of the audiometric gain. Patients presented either cholesteatomatous otitis sequelea or any preliminary status (retraction pockets) with attical impairment. PMID- 7486708 TI - [Malformation of the internal ear and recurrent meningitis: radio-surgical correlations]. AB - Authors report one case of bilateral inner ear malformation in a 3 years-old child with congenital sensorineural hearing loss, recurrent meningitis and cerebro-spinal fluid leakage. This anomaly is a pseudo-Mondini's malformation with a meningocele through the oval window and a fistula of C.S.F. of the round window. A close radiological and surgical correlation support some physiopathological hypothesis about fistula between sub arachnoid space and middle ear. Authors discuss the place of high resolution computed tomography in case of congenital sensorineural learning loss. PMID- 7486706 TI - The kinase-dependent function of Lck in T-cell activation requires an intact site for tyrosine autophosphorylation. AB - The cytoplasmic protein tyrosine kinase p56lck (Lck) has important signaling roles in T-cell development and activation. We have mutated the two known regulatory tyrosine residues of CD4-associated Lck and examined the effects on its kinase-dependent function in an antigen-specific CD4-dependent T-cell hybridoma. Substitution of phenylalanine for the negative regulatory tyrosine-505 within a CD4/Lck chimera resulted in a slightly increased response to antigen, whereas mutation of the major in vitro autophosphorylation site (tyrosine-394) completely abolished the kinase-dependent function of Lck. Even though its kinase activity was only slightly affected, the F394 mutant behaved similarly to a catalytically inactive chimeric protein. Cross-linking of the F505 mutant, but not of wild-type Lck or F394 mutants, resulted in tyrosine phosphorylation of multiple cellular proteins. Although the pattern of tyrosine phosphorylation resembled that observed upon T-cell receptor cross-linking, there was no induction of interleukin-2 synthesis upon cross-linking of the chimeric protein. These results suggest that the activity of the Lck kinase domain in vivo is controlled by dephosphorylation at the negative regulatory site and phosphorylation at the positive regulatory (autophosphorylation) site. Additionally, our data show that the specific kinase activity of Lck towards an artificial substrate need not correlate with its ability to phosphorylate cellular proteins or its biological function. PMID- 7486710 TI - A lip burn as a consequence of laser laryngeal surgery. AB - Ignition and subsequent combustion of a gum guard, during laser surgery for tracheal stenosis, resulted in a superficial burn to the lower lip. We present a case report and discuss measures that can be taken to avoid such a complication. PMID- 7486712 TI - [Local evaluation of cancers of the piriform sinus]. PMID- 7486711 TI - [The thesis of Reininger (1722) in the history of pituitary sinuses]. AB - The thesis written by Sebastian Reininger in 1722 was conserved by Albert de Haller, the renowned scholar who collected interesting medical publications in the 18th century. He included Reininger's work on the pituitary sinuses in his Disputationes anatomicae. This thesis is probably the first published work dealing exclusively with the sinuses and their anatomy, function and pathology. There are also some comments on therapy. It was thus written before thesis by Ludolph Heinrich Runge (1750) often cited as the earliest work in this field. The Reininger dissertation is based on a personal series of cranial dissections and is completed with references to other authors of the same period. It is an interesting synopsis of the knowledge of pituitary sinuses at the beginning of the 18th century. PMID- 7486713 TI - [Objective vocal evaluation of dysphonia by simultaneous measurement of acoustic and aerodynamic parameters with the EVA device]. AB - Authors present the analysis of the voice in 88 normal subjects and 159 dysphonic patients by simultaneously measurements of Jitter (acoustical short-term indicator of aperiodicity) and "Glottal Leakage" (aerodynamic parameter calculated by the ratio of Oral Airflow and Intensity). All the measurements were performed, with EVA apparatus, on the most stable part of a steady-state vowel /a/. The reference method was the rating by a jury along a global perceptual scale. Statistical tests confirm that Jitter and "Glottal Leakage" are relevant to evaluate the quality of the voice. Discriminant analysis allows to find out a fairly-good rate of discrimination of 63.1% (Randomly rating would have been 25%) The authors conclude on the interest of such a protocol and present suggestions to improve it by taking in count phenomena during the vocal attack and middle term variations. PMID- 7486709 TI - [Primary cutaneous neuroendocrine carcinoma (Merkel tumor). Clinical and therapeutic update apropos of 4 cases]. AB - The clinical and pathological features of 4 cases of cutaneous neuroendocrine carcinoma are reported. There are unusual malignancies mostly observed in elderly, the prognosis is poor by loco-regional aggressivity and disseminated metastases. They are diagnosed by a pathologist: ultrastructural studies and immunohistochemistry support the diagnosis. A combination of a wild local excision, node dissection and post-operative radiation is recommended. PMID- 7486714 TI - [Surgical anatomy of the parathyroid glands. Apropos of 200 cases. Practical implications]. AB - An operative procedure aimed at decreasing the risk of post-surgery hypothyroidism was proposed on the basis of parathyroid gland surgical anatomy described from a retrospective study of 200 thyroidectomies performed by the same operator It is almost always possible to visualize 1 or parathyroids during thyroid procedures. This fact makes it possible to reduce the rate of definitive hypoparathyroidism to less than 5% in surgery for invasive cancer of the thyroid and to 1% for non-invasive cancers requiring total thyroidectomy and uni or bilateral mediastinorecurrent curettage. PMID- 7486715 TI - [Analysis of results of pharyngotomy in the surgical treatment of sleep apnea syndrome. Apropos of 150 cases]. AB - The indications and limitations of simple veloamygdalotomy as surgical cure for sleep apnoea were analyzed on the basis of results obtained in the first 150 cases treated prospectively by pharyngotomy. Clinical and polysomnographic results were analyzed as possible factors predicting success or failure. With a success rate of 80%, pharyngotomy is a simple and effective treatment for patients with minor forms of sleep apnoea (initial apnoea/hypopnoea index < 20) and no severe obesity. It appears unreasonable to propose isolated pharyngotomy if the initial index is < 30 since the success rate in the cases is only 27%. Nasal repermeation does not improve overall results significantly. The lack of patient compliance to diagnostic and therapeutic modalities is an unavoidable reality due to human, social and economic implications. PMID- 7486717 TI - [Local cutaneous necrosis after injection of interferon beta]. AB - INTRODUCTION: Skin necrosis at the site of interferon injections is unusual. With beta-interferon, it has been observed with high doses ranging from 90 to 180 millions daily. CASE REPORT: Is reported a case of abdominal skin necrosis following 6 months treatment with beta-interferon injections, 9 millions 3 times a week, in a 56 years-old man with chronic type C hepatitis. Histological analysis showed fibrin thrombosis of deep dermal vessels. There was no coagulation abnormality. DISCUSSION: The mechanisms of the interferon induced skin necrosis are discussed. PMID- 7486716 TI - [Association of lichen sclerosus and monomelic scleroderma]. AB - INTRODUCTION: Coexistence of lichen sclerosus and scleroderma is well demonstrated. However, clinical and histological lesions of lichen sclerosus and scleroderma, in a linear pattern, do not seem to have been reported. CASE REPORT: We report the case in a six years old boy, of both lichen sclerosus and linear scleroderma lesion that involved his left lower limb. Immunological and inflammatory investigation was normal or negative, as well as, bone and muscles analyze of the leg. Further outcome was benign. DISCUSSION: This new case confirms the possibility of an association, inside a same limb, of lichen sclerosus and localized scleroderma, and supports the concept of a common etiological process in these two diseases. In an other hand, it could reflect the possibility of two distinct clinical expressions of the same pathogenic process. PMID- 7486718 TI - [Linitis plastica disclosed by cervical cutaneous metastasis]. AB - INTRODUCTION: A case of a cutaneous metastasis as a first sign of a linitis plastica is reported. CASE REPORT: A 57 year-old man presented for a cervical infiltrated skin plaque. Histological examination and immunohistochemical staining gave the diagnosis of metastasis probably of gastrointestinal origin. Gastric endoscopy and biopsy confirmed the diagnosis of a linitis plastica. DISCUSSION: Cutaneous metastases from gastric carcinoma are uncommon and exceptionally the first sign of the disease. Their clinical and histological aspects are reviewed. PMID- 7486719 TI - [Targetoid hemosiderotic hemangioma]. AB - INTRODUCTION: Targetoid hemosiderotic hemangioma (THH) is a new vascular disorder, rarely reported. The lesion presents as violaceous papule surrounded by an ecchymotic ring usually measuring less than 1 cm in diameter. Epithelioid endothelial cells with a hobnail appearance, vascular papillary projections and hemosiderin deposits are the histological hallmarks of this entity. OBSERVATION: We report a case fulfilling these criteria. Particular morphologic features were the presence of a very prominent lymphocytic infiltrate, with numerous intraluminal lymphocytes closely associated with the neoplastic endothelial cells, scarcity of siderophages, rought of retiform pattern, predilection of newly formed vessels for muscles of hair erection and eccrine ducts. DISCUSSION: Having regard to these worrisome histologic features the differential diagnosis between THH, Kaposi's sarcoma and newly recognized entities such as lymphangioendothelioma and retiform hemangioendothelioma is discussed. PMID- 7486720 TI - [Alternaria tenuissima plurifocal cutaneous infection]. AB - INTRODUCTION: Multiple skin localization of phaeohyphomycoses are rare and are encountered in immunodepressed subjects. We report a case due to Alternaria tenuissima in a patient given corticosteroids and IgA deficiency. CASE REPORT: A 73-year-old man presented with ulceronecrotic lesions on the two lower limbs which had developed over 15 days and poor general health. He had been taking corticosteroids for retroperitoneal fibrosis for 15 days. Skin biopsies demonstrated monoclonal dysglobulinaemia with IgG kappa and IgA deficiency. Itraconazol treatment was successful. DISCUSSION: Identification of Alternaria is in a skin sample, it not sufficient to determine its pathogenic nature. Strict criteria, especially histological criteria, are required. Most cases of cutaneous alternariosis are localized infections due to direct inoculation. They are seen in healthy patients or more often in immunodepressed subjects. Disseminated skin disease is rare and has been reported in 2 highly immunodepressed patients. Our patient was taking corticosteroids, but his deficiency state in IgA may have played a role in the development of the mycosis. Itraconazol appears to be effective treatment. PMID- 7486721 TI - [A case for diagnosis: Wells syndrome]. PMID- 7486722 TI - [A case for diagnosis: syndrome of hypersensitivity to carbamazepine]. PMID- 7486723 TI - [Bullous eruptions induced by phototherapy]. PMID- 7486725 TI - [Monthly question: what should be done in diffuse alopecia in women?]. PMID- 7486724 TI - [Cutaneous immunoallergic reactions caused by vaccines]. PMID- 7486726 TI - [Intimate journal]. PMID- 7486727 TI - [Dyshidrotic pemphigoid]. AB - INTRODUCTION: Clinical expression of pemphigoid can sometimes be misleading. Localized dyshidrosis is rarely reported in the literature. CASE REPORT: We observed 4 cases of pemphigoid with unusual dyshidrosis lesions. Skin biopsy was unspecific in both cases. Immunofluorescence gave the diagnosis in both. One case responded to low-dose general corticosteroids associated with local applications of corticosteroids. The other patients required high-dose oral corticosteroids. CONCLUSION: Our cases illustrated the misleading dyshidrosis nature of certain pemphigoids and the need for repeated skin biopsies with immunofluorescence in case of recent development of dyshidrosis in elderly subjects. As in the literature, and despite the localized nature of the lesions, general corticosteroid treatment appears necessary in many cases. PMID- 7486728 TI - [Photobiological study of lupus erythematosus]. AB - BACKGROUND: Although cutaneous photosensitivity is one of the major criteria used for the diagnosis of systemic lupus erythematosus, this criterion is not precise. OBJECTIVE: To evaluate the interest of photobiological exploration in patients with cutaneous lupus erythematosus, and to compare the results to clinical photosensitivity. METHODS: Nineteen patients consecutively seen in one year, have been tested using a standardized method. RESULTS: Clinical photosensitivity was present in 16 cases. The clinical presentation of cutaneous disease was classified in 3 groups: acute, subacute, and chronic. One patient with no history of photosensitivity had positive photobiological tests, and five patients had negative tests though they experienced clinical photosensitivity. Positive phototests with UVA and UVB were present in 6/8 subacute lupus cases, 3/6 acute lupus cases, 3/5 chronic lupus cases. Histological aspect of cutaneous biopsies from phototest was not characteristic of lupus. Minimal erythematous doses was comparable in lupus group and in controls. No correlation between positivity of phototests and the presence of systemic involvement or anti-Ro/SSA antibodies was established. CONCLUSION: Phototesting is useful to assess photosensitivity in lupus patients. PMID- 7486729 TI - [Lichenoid eruptions of the mouth mucosa induced by lithium carbonate. 2 cases]. AB - INTRODUCTION: Several drugs can induce lichenoid lesions. CASE REPORT: We observed two cases of lichenoid lesions of the buccal mucosa due to lithium carbonate. Drug withdrawal caused clearing. DISCUSSION: These cases emphasize the rare nature of this complication. To date only one lichenoid stomatitis has been reported in the literature. PMID- 7486731 TI - [Pyoderma gangrenosum, subcorneal IgA pustulosis and recurrent neutrophilic pleural and pulmonary diseases in a patient with IgA gammopathy]. AB - INTRODUCTION: The term "neutrophilic dermatosis" is normally used to cover a number of dermatoses characterized by their response to corticotherapy or to drugs effective on the neutrophilic polynuclear. Neutrophilic dermatosis was frequently associated with hematologic disease. We study the case of a neutrophilic dermatosis accompanied by a pulmonary involvement and then review the literature on the reported cases of neutrophilic dermatosis with extra cutaneous localisations. CASE STUDY: A woman aged 54 displayed a pyoderma gangrenosum successively followed by a pustular dermatosis (related to subcorneal pustular dermatosis of Sneddon-Wilkinson) and by a pulmonary involvement not caused by any underlying infection. The laboratory results revealed a benign IgA gammaglobulin. The use of corticotherapy and a relay by dapsone led to the gradual disappearance of the cutaneous and pulmonary lesions. DISCUSSION: Pulmonary involvement have been observed in the Sweet syndrome and in the pyoderma gangrenosum. The neutrophilic lung is diagnosed once a tumorous or infectious cause has been eliminated and on the basis of the response to the cortisone treatment. CONCLUSION: The link between pulmonary involvement and neutrophilic dermatosis is sometimes mentioned but is in fact often difficult to confirm. The positive reaction to corticosteroids could be a diagnostic test. PMID- 7486730 TI - [Necrotic purpura of the head disclosing Churg-Strauss syndrome]. AB - INTRODUCTION: Among the cutaneous manifestations of the Churg-Strauss syndrome, palpable purpura of the extremities is the most common. Although they are not specific, sub-cutaneous nodules of the scalp are the most distinctive lesions. CASE REPORT: A 58 year-old man, with a 7-years history of asthma, developed profuse cutaneous lesions involving the face and the scalp, initially erythematosus and vesiculous, then purpuric and necrotic which led to the diagnostic of Churg-Strauss syndrome. COMMENTARY: Cutaneous lesions occur frequently during the Churg-Strauss syndrome and may reveal the disease as in our patient. However, the initial clinical presentation was uncommon with erythematosus and vesiculous lesions mimicking primary herpes infection. Cutaneous biopsy was useful leading to an early diagnosis. PMID- 7486732 TI - A golfer who changed the world. PMID- 7486733 TI - Toxic epidermal necrolysis: a review and report of the successful use of Biobrane for early wound coverage. AB - Toxic epidermal necrolysis (TEN), is an exfoliative dermatological disorder of unknown cause. A patient with TEN loses epidermis in sheet-like fashion, leaving extensive areas of denuded dermis that must be treated like a large, superficial, partial-thickness burn wound. Methods of coverage described in the English literature over the last decade include the use of several dressings such as fresh-frozen or cryopreserved cadaver allograft, porcine xenograft, and amnionic membrane. Successful use of the biosynthetic dressing, Biobrane, has been described after burn injuries and Stevens-Johnson syndrome; however, its use in TENS has not. We present three patients with TEN treated successfully in our burn center over the past 12 months using Biobrane. The patients were men aged 20, 58, and 77 years, with 58% to 95% total body surface area slough. Diagnosis was confirmed by skin biopsy on admission, and drug ingestion was implicated in each instance. Each patient was thoroughly debrided in the operating room shortly after admission, and denuded areas were covered with Biobrane within 24 to 48 hours of admission. Biobrane demonstrated greater than 90% adherence by 48 hours, and no wound sepsis occurred. Each patient demonstrated epithelialization within 9 days. Patients were ambulatory at 72 hours. Corticosteroids and prophylactic antibiotics were avoided. Enteral nutritional support and aggressive septic surveillance was routine. Hospital stay was between 13 and 30 days without mortality. Early use of Biobrane in patients with TEN appears to provide a reasonable means of wound coverage. PMID- 7486734 TI - Significant hemangiomas and vascular malformations of the head and neck: clinical management and treatment outcomes. AB - The treatment dilemmas and our institutional experience with head and neck hemangiomas and vascular malformations are reviewed. Fifty-eight patients (38 with hemangiomas, 20 with vascular malformations) aged from newborn to 78 years were treated from 1975 to 1993. Eighty-seven percent of patients provided up-to date follow-up, yielding a median follow-up interval of 7.18 years from intervention. Only significantly sized tumors (minimum diameter > 4 cm) were studied. Sodium tetradecyl sulfate sclerotherapy (SDS) was a sole therapy in 12 patients (all with hemangiomas) and was used in combination with surgical ablation in 22 patients (16 with hemangiomas, 6 with vascular malformations). The minority of tumors from both categories were completely removed with ablative surgery (six hemangiomas [23%], eight vascular malformations [44%]). Significantly debilitating tumor postsurgical recurrence, which led to mortality in some patients, was unique to vascular malformations with high flow on the basis of increased arterial to venous shunting. Less alarming but significant functional and cosmetic challenges are created in primarily postadolescent patients with incompletely involuted hemangiomas and low-flow vascular malformations of the head and neck. We have had a favorable experience with the combined application of sclerotherapy and conservative ablative surgery in this group. Patient survey implied satisfaction with treatment in the majority: Nine indicated they benefited significantly (17%); 37 indicated they were generally improved (71%); and 6 stated they did not benefit (12%). Tumor-specific philosophy on ablative therapy, recurrence patterns, and associated complications are presented. PMID- 7486736 TI - Argon and Q-switched yttrium-aluminum-garnet laser treatment of nevus of Ota. AB - Twenty patients with nevus of Ota have been treated with the argon laser, Q switched yttrium-aluminum-garnet (YAG) laser, or a combination of both lasers. The argon laser was originally used from 1975 to 1993 and has now been replaced by the Q-switched YAG laser. The Q-switched YAG laser mechanism of action is selective photothermolysis in which the pulse width of the laser beam is very short, thus allowing the melanocyte chromophore target to dissipate its heat without burning adjacent normal tissue. Satisfactory fading without skin scar or texture change was observed in 10 patients in this series who had completed treatment; treatment is in progress in the remaining 10 patients. Six patients who were initially treated with argon and then subsequently were treated with Q switched YAG laser demonstrated improved results. PMID- 7486735 TI - Intralesional laser photocoagulation-steroids as an adjunct to surgery for massive hemangiomas and vascular malformations. AB - Massive hemangiomas and vascular malformations have been treated with lasers and adjunctive measures. Intralesional laser photocoagulation involves the direct penetration of a tumor by a yttrium-aluminum-garnet laser fiber with photocoagulation at various depths and levels to produce thromboses and controlled coagulation necrosis, thus shrinking and blanching the malformation. Seven children have benefited from intralesional laser photocoagulation as well as steroids, embolization, and excisional surgery. Intralesional laser photocoagulation has contributed to the initial shrinkage and fibrosis of these massive vascular tumors in preparation for surgery or as the only treatment in some cases. PMID- 7486737 TI - Animal study of skin resurfacing using the ultrapulse carbon dioxide laser. AB - Ultrapulse carbon dioxide laser ablation of rabbit skin was investigated. Macroscopic and microscopic appearance of the ablation site was evaluated for 1 month. The Ultrapulse carbon dioxide laser was set in the ultrapulse mode and the parameters selected were 250 mJ of energy per pulse and 2 W average power with the 3-mm spot size collimated handpiece. Macroscopic investigation showed that ultrapulse carbon dioxide laser ablation removed rabbit skin precisely and bloodlessly without char formation. Skin re-epithelialization without scarring and perfect hair growth restoration within 3 weeks were impressive. Microscopic investigation demonstrated a layer of tissue thermal necrosis measured as 70 +/- 10 microns. During the first 2 weeks, laser ablation wound re-epithelialization begins with multiplying epidermal layers (from 2-3 cell layers to about 13 to 15 cell layers). There were marked fibroblastic reactions and capillary congestion within papillary dermis. By the end of 4 weeks, the thickened epidermal layer appeared normal. The repaired papillary dermal collagen showed a relative compact configuration and greater cellularity in Masson stain was found 2 to 3 weeks later and also was near normal 4 weeks after laser treatment. These findings suggest that Ultrapulse's delivery of short-duration, high-energy pulses may provide a new approach (laser dermal ablation) to damage selectively the epidermis and papillary dermis of rabbit skin in one single-pulse impaction with uneventful wound healing within 1 month. PMID- 7486738 TI - Thermographic mapping of perforators and skin blood flow in the free transverse rectus abdominis musculocutaneous flap. AB - There is no ideal method for preoperative or intraoperative mapping of cutaneous perforators or for postoperative monitoring of blood flow in cutaneous flaps. To study the suitability of thermography for the mapping and monitoring of free transverse rectus abdominis musculocutaneous (TRAM) flaps for breast reconstruction, we performed thermography pre-, intra-, and postoperatively (eight patients). The temperature of the TRAM flap increased during the induction of anesthesia and was still higher than normal on the first and second postoperative days (p < 0.05). During the operation, the flap cooled, reaching its minimum temperature (3.62 +/- 0.6 degrees C below phase 2, p < 0.05) after ligation of both pedicles of the flap. When blood flow had been re-established, all parts of the flap warmed rapidly. The locations of perforators ("hot spots") could be seen before, during, and after the operation. However, induction and cutting of both pedicles made the flap isothermic, and the perforators disappeared temporarily. Thermography is a potential method of mapping cutaneous perforators pre-, intra-, and postoperatively and of monitoring the flaps at bedside. The method is easy to use and the outcome can be seen immediately. Our results also showed that the temperature (blood flow) in free TRAM flaps is higher than in the tissue in its original position. PMID- 7486739 TI - Effect of anticoagulation and inhibition of platelet aggregation on arterial versus venous microvascular thrombosis. AB - On the basis of clinical empirical observations and classic theory of platelet aggregation, microvascular surgeons have held that platelet aggregation is more responsible for arterial thrombosis and fibrin clot formation is more responsible for venous thrombosis. This theory was tested in two models of thrombosis, one created in arteries and the other in veins, using systemic antithrombotic agents. The models were pair matched in rats, creating a crush-avulsion injury to the femoral artery on one leg and a crushing injury to the femoral vein on the contralateral leg. There were four treatment groups (n = 15 per group): systemic heparin, aspirin and dipyridamole, both sets of agents, and dual controls (vehicle treated). One-day patency rates in the controls were 13.4% and 0% for the arterial and venous models, respectively. Heparin treatment caused an increase in arterial patency to 73.3% but had no effect on venous patency (6.7%). Aspirin and dipyridamole had a modest effect on venous patency (40%) but had no effect on arterial patency (6.7%). Administration of both sets of agents led to 100% and 86.7% patency in the arterial and venous models, respectively. Patency rates were maintained at the 7-day evaluation in the arteries; in contrast, all thrombosed veins were found to be patent 7 days postoperatively (100% patency in all groups). These findings are in contrast to classic thinking about microvascular thrombosis, but this reverse effect may be unique to the rat. The high rate of recanalization in thrombosed veins indicates a need for caution in performing experimental studies of patency in rat veins. PMID- 7486740 TI - Squamous cell carcinoma growth in irradiated tissue: a murine model for quantitative assessment of treatment. AB - Locoregional recurrent and distant metastases from squamous cell carcinomas, despite multimodality therapy, remain troublesome clinical realities. Discrepancies in success rates of various surgery and radiation treatment regimens dealing with these problems are confusing to the clinician attempting to recommend the most beneficial treatments. There is a need for an experimental model to assess therapeutic effectiveness quantitatively from which guidelines for developing clinical trials may be suggested. In this study, we provide such a model. We injected DBA-2 mice with defined numbers of KLN-205 squamous carcinoma cells to obtain baseline growth characteristics; 216 animals had no previous irradiation. The remaining 131 received 30 Gy irradiation to the right leg 50 days before injection of the tumor cells. Tumor incidence, growth and number, and location of tumor metastasis were determined in both previously irradiated and nonirradiated groups. The data demonstrate a growth-retardant effect on tumor groups by the previous irradiation (tumor bed effect). The data also show that the incidence of hematogenously spread metastases was more frequent in mice in which tumors developed in previously irradiated tissue than in mice with tumors in nonirradiated tissue. PMID- 7486741 TI - Calcitonin gene-related peptide increases microcirculation after mechanically induced ischemia in an experimental island flap. AB - Vasoconstriction, as a result of mechanical manipulation of blood vessels during microsurgery, may produce a decrease in blood supply and endanger flap viability. A study was undertaken to determine the effects of the topical vasodilator calcitonin gene-related peptide on the microcirculation of flaps after mechanically induced ischemia. A neurovascular island flap based on the superficial epigastric vessels was raised in 42 rats. Blood cell flux in the flap was recorded continuously with a laser Doppler flux meter. The feeding artery was pinched to induce vasospasm, and different concentrations of calcitonin gene related peptide (10(-7), 10(-8), 10(-9), 10(-10) mol) or a control of sodium chloride 0.9% was applied topically to relieve the ischemia. Results showed that calcitonin gene-related peptide at a concentration of 10(-7) mol significantly shortened the time to reach 50% of the original blood cell flux values (270 +/- 123 seconds) and significantly increased the number of flaps in which the blood cell flux values were restored to prestress levels within 30 minutes. The data support the conclusion that, in this model, topical calcitonin gene-related peptide at the concentration of 10(-7) mol was effective in promoting recovery of the microcirculation after mechanically induced ischemia, without the adverse effects associated with other commonly used vasodilators. PMID- 7486742 TI - Microvascular transplantation of cryopreserved knee joints. AB - Successful transplantation of cryopreserved complex tissues has not yet been reported. We wanted to test whether microvascular transplantation of cryopreserved knee joints can be achieved in rats. Twenty-five knee joints were cryopreserved and then microsurgically transplanted to the groin of inbred recipient animals. Ten fresh knee joints were transplanted as controls. One week after transplantation, the transplants were evaluated in situ for pedicle patency and tissue perfusion and then harvested for histology. In all surviving rats (n = 21), the cryopreserved knee joints were well perfused immediately after transplantation. At harvest, 14 knee joints appeared viable with a patent pedicle, whereas 6 were questionably viable and demonstrated arterial thrombosis. Histologically, the viable joints revealed an intact bone and cartilage matrix; however, the number of viable chondrocytes and osteocytes was reduced. Additionally, endothelial cell plugs in some areas of the capillary bed led to scattered microinfarctions despite a patent vascular pedicle. The nonperfused transplants showed infarction. In conclusion, we have demonstrated successful microvascular transplantation of cryopreserved knee joints with reduced short term viability. Long-term studies must determine whether full recovery occurs. PMID- 7486744 TI - Free fat autotransplantation for the cosmetic treatment of first web space atrophy. AB - The results of free fat autotransplantation for the correction of first web space atrophy were reviewed for 25 patients who sustained war injuries resulting in ulnar nerve palsies. In anticipation of partial graft resorption, a 30% overcorrection of the first web space deformity was performed compared with the normal hand. Average follow-up was 5 years. All patients were satisfied with their cosmetic results. Whereas 21 patients (84%) demonstrated less bulk when compared to their normal side, 4 (16%) retained slight overcorrection of the web space. After 1 year of follow-up, there was no further evidence of graft resorption. There were no long-term complications. We conclude that free fat autotransplantation for the cosmetic correction of atrophy of the first web space is a safe and effective procedure. Although graft resorption was encountered, overcorrection provided patients with dramatic cosmetic improvement. PMID- 7486743 TI - Quantification of the relationship between dynamic grip strength and forearm rotation: a preliminary study. AB - Clinically, the measurement of grip strength is a static measure. It is performed in a standardized posture with the arm at the side, elbow flexed to 90 degrees, and the wrist-forearm positioned in neutral. Many functional activities require dynamic grip strength when an individual moves the wrist-forearm while simultaneously gripping an object. This preliminary study investigated dynamic forearm rotation with isometric grip strength over real time. Thirty wrists from 15 normal subjects were measured with a tilt sensor attached to a strain gauge dynamometer. Each subject underwent randomized trials of maximally gripping the dynamometer while rotating the forearm over a 10-second period. A multiple regression model was fit for the parameters of extremity, forearm rotation order, fatigue, and loss of strength with supination and pronation. For each parameter, the mean and standard error of its estimate were computed and compared with zero. The significant effects (p = 0.05) of interest were for fatigue and the loss of grip strength from 70 degrees of supination to the end range of supination. Comparisons to existing static grip strength literature were made. We concluded that there is a decrease in grip strength over time as a result of fatigue, there is a decrease in grip strength at supination angles greater than 70 degrees. There is no decrease in grip strength resulting from supination angles less than 70 degrees, nor is there a decrease in grip strength with pronation when measured dynamically. It is suggested that there may be a difference in static and dynamic GS measurements related to forearm rotation. PMID- 7486745 TI - Synovial metaplasia of a periprosthetic capsule surrounding a polyurethane foam breast prosthesis. AB - A 43-year-old woman underwent two-stage bilateral reconstruction mammoplasty using silicone-filled, polyurethane foam-covered breast implants. The permanent implants were later removed. The periprosthetic capsule demonstrated synovial metaplasia by light and electron microscopy. Synovial metaplasia is believed to be induced by the physical and chemical characteristics of the prostheses and may have important clinical implications in reducing capsular contracture and increasing host acceptance of implantable biomaterials. PMID- 7486746 TI - Atypical Mycobacterium and breast surgery. AB - An unusual case of atypical mycobacterium infection following breast ptosis correction is described. This was successfully treated with a combination antibiotic regime. Breast surgery and atypical mycobacterium is discussed with reference to diagnosis management and suggested treatment regimes. PMID- 7486747 TI - Surgical management of life-threatening epistaxis in Osler-Weber-Rendu disease. AB - Hereditary hemorrhagic telangiectasia is a systemic disorder of blood vessels. It often leads to severe epistaxis that is refractory to conventional therapy. Three patients with severe epistaxis caused by hereditary hemorrhagic telangiectasia unresponsive to laser coagulation and to septal dermoplasty and requiring multiple transfusions underwent extirpation of nasal mucosa through a rhinotomy approach, with a median forehead flap and split-thickness skin graft coverage of the mucosal defects over the floor and lateral walls of the nose. All patients had reduced frequency and severity of bleeding without any need for transfusions during follow-up periods of 6 months, 3 years, and 5 years, respectively. Forehead flaps resulted in minimal morbidity. Partial nasal obstruction and a forehead scar were the main complaints. Surgical technique, complications, and alternative therapy are discussed. PMID- 7486749 TI - Secure dressing after nipple-areola reconstruction. AB - Firm contact to the underlying recipient bed and immobilization are necessary for survival of an areolar skin graft, yet compression of the nipple must be avoided when performing nipple-areola reconstruction. A simple dressing technique is described that simultaneously accomplishes both goals and can be carried out with commonly available materials. PMID- 7486748 TI - Use of type III venous flaps: single- and multistaged procedures. AB - Two cases are presented in which the principles of type III (arteriovenous) venous flaps were applied in single- and multistage procedures. In the first case, the flap was used for correction of a post-thermal burn adduction contracture of the first web space of the hand as a one-stage microvascular free flap and in the other for the resurfacing of a post-traumatic defect on the dorsum of the foot as a two-stage transfer. A brief review, along with the technique, advantages, mode of survival, and feasibility of using type III venous flaps, is presented. A simple classification of type III venous flaps is also suggested. PMID- 7486750 TI - The confusing ethics of mismanaged care. PMID- 7486751 TI - Re: Treatment of vitiligo with melanocytic grafting. PMID- 7486752 TI - Re: Risk of dissemination of cancer to flap donor sites during immediate reconstructive surgery. PMID- 7486753 TI - Oral cancer: a serious and growing problem. PMID- 7486754 TI - Surgical management of carotid stenosis. PMID- 7486756 TI - An audit of one surgeon's experience of oral squamous cell carcinoma using computerised malignancy database. AB - The results of an audit of a single surgeon's experience of oral squamous cell carcinoma over a 5-year period is reported and the benefits of adopting a prospective computerized malignancy database outlined. Oral cancer occurred mainly in the 7th decade of life, women were on average 5 years younger than men at presentation. Men were affected 1.7 times as often as women and presented with more advanced disease than women. The mean delay from referral to histological diagnosis was 6.4 days and the mean delay from diagnosis to treatment was 25.8 days. The majority of patients consumed alcohol and smoked tobacco, but a significant minority were lifelong nondrinkers and non-smokers. While the incidence of most site-specific oral cancers were similar to the national averages the incidences of alveolar and floor of mouth cancers were higher than expected. The TNM stage at presentation was an unreliable prognostic indicator of future disease progression and outcome. Auditing treatment and outcome is a major component of the recent proposals to centralise the provision of cancer services in dedicated cancer centres and units (1). Audit can only be effective if it is based on current, reliable and accurate data and this can only be achieved by the use of a computerised malignancy database. PMID- 7486755 TI - Survival, metastasis and recurrence of oral cancer in relation to pathological features. AB - The outcome of 123 patients with squamous cell carcinoma of the oral/oropharyngeal mucosa treated by primary radical surgery and simultaneous neck dissection and followed up for a maximum of 5 years is reported and related to the pathological features of the tumour. Actuarial survival analysis showed the overall 1-year survival probability was 84%, falling to 69% at 2 years, and 65% at 5 years. For patients without lymph node metastasis, the corresponding survival probabilities were 95%, 86% and 86%, respectively; and for patients with metastasis, 71%, 52% and 44%, respectively. In all, 30 patients (24%) died of their oral cancer; 16 (13%) of local recurrence, 10 (8%) of regional recurrence and 4 (3%) of systemic metastases. In addition to lymph node metastasis, survival was related to the site and stage of the primary tumour, histological pattern of invasion, status of the resection margins and, for patients with metastasis, the number and anatomical level of positive nodes and the presence and extent of extracapsular spread. These pathological features are important indicators of tumour behaviour and should be incorporated into protocols for assessment of prognosis. PMID- 7486758 TI - Oesophageal epiphrenic diverticulum: an unusual presentation and review. AB - Epiphrenic pulsion diverticula are rare and often asymptomatic. We describe a case presenting in an unusual fashion, and review the controversy over the management of this condition with regard to the requirement for myotomy and antireflux surgery. We believe that both procedures are necessary, but believe that both procedures are necessary, but optimal management strategies are unlikely to be resolved as the rarity of the condition precludes largescale prospective studies. PMID- 7486757 TI - Radiological predictors of response to renovascular reconstructive surgery. AB - The aim of this study was to identify preoperative angiographic criteria which could be used to predict the response to renovascular reconstruction. A cohort of 49 patients who survived for more than 6 months after renovascular reconstruction was studied. All preoperative angiograms were reviewed independently and the renal size, degree of stenosis and the length of the lesion recorded. The response of blood pressure and renal function to surgery were correlated with the angiographic findings. We found that 58% of patients had long-term improvement in blood pressure control, though the response was significantly better where both kidneys were >12 cm in size (82%) compared with when both kidneys were < 12 cm (25%), P < 0.02. The degree of stenosis, while showing a trend towards a better blood pressure response with increasing stenosis, was not a statistically significant factor. None of the angiographic criteria examined could be used to predict the response in renal function. We have shown that the preoperative renal size is the only angiographic factor that may have some role in predicting the response of blood pressure to renovascular reconstruction. PMID- 7486760 TI - Anterior cruciate reconstruction with the ABC carbon and polyester prosthetic ligament. AB - Twenty-one knees with isolated chronic deficiency of the anterior cruciate ligament have been treated by reconstruction using the ABC carbon and polyester bioprosthetic composite ligament; 16 were reviewed at an average of 34.6 months. The Lysholm score indicated a good or excellent result in nine knees (56%) with a score of 84 points or more, a fair result in one (65 points or more) while four patients (25%) had a poor result ( < 65 points). The ligament failed in two cases. There were two failures in one patient, and one ligament was removed because of deep sepsis. In the remainder, 14 patients, a positive pivot shift was identified in 13 preoperatively and in three patients at review. These results are similar to those reported recently by others. We have stopped using the ABC carbon and polyester ligament in isolated anterior cruciate deficient knees. PMID- 7486759 TI - Severe hyponatraemia in elderly patients: cause for concern. AB - An increased incidence of severe hyponatraemia in an orthopaedic unit led to an investigation into the possible causes. Routine perioperative administration of 'hypotonic' intravenous fluid to an elderly population emerged as the most important aggravating factor. Care is necessary in prescribing intravenous fluid regimens in order to avoid potentially serious complications. PMID- 7486761 TI - Current UK opinion on thromboprophylaxis in orthopaedic surgery: its use in routine total hip and knee arthroplasty. AB - We surveyed all surgically qualified members of the British Orthopaedic Association as to their views on thromboprophylaxis in routine total hip and knee replacement. The response rate was 92%. In total hip replacement, 13-15% of surgeons used no method of prophylaxis at all, 18-22% did not use a mechanical method of prophylaxis, while 26% did not use a pharamacological agent; 36% believed low-dose anticoagulant thromboprophylaxis to be a medicolegal necessity while 47% disagreed with this. In total knee replacement, 18-33% of surgeons used no method of prophylaxis at all, 20-26% did not use a mechanical method of prophylaxis, while 54-58% did not use a pharmacological agent; 12 believed low dose anticoagulant thromboprophylaxis to be a medicolegal necessity, while 64% disagreed with this. We conclude that mechanical methods remain a popular method of thromboprophylaxis and pharmacological methods, while commonly used, are not seen as a medicolegal requirement in lower limb arthroplasty and are used less often in total knee replacement. PMID- 7486762 TI - Predicting the prognosis after proximal femoral fracture. AB - In 338 patients with proximal femoral fractures, age, sex, residence and mobility on admission were found to predict mortality, mobility and housing requirements after 1 year. Application of these data should permit better targeting of rehabilitation and earlier initiation of rehousing arrangements after proximal femoral fracture. PMID- 7486763 TI - Acute appendicitis: does removal of a normal appendix matter, what is the value of diagnostic accuracy and is surgical delay important? AB - A prospective study with long-term follow-up was undertaken of 248 patients (137 males, median age 18 years (range 6-81 years), undergoing emergency appendicectomy during a 12-month period. Acute inflammation was present in 182 patients (73.4%) (males 86.1%, females 57.8%; P < 0.001). Before surgery, the positive predictive value of diagnostic accuracy was 82.0% (males 91.2%, females 67.7%). Delaying surgery did not significantly increase the proportion of perforated appendices (22.0%), hospital stay, or frequency of postoperative complications (overall 49.6%). Hospital complications were significantly more common among patients with a perforated appendix. There was no significant difference in the complication rate between patients with or without appendicitis while in hospital, during the first 18 months after operation or 8 years after operation. At 18 months, 17 of 238 patients (7.1%) continued to experience their original pain. After 8 years the original pain was still present in 10 of 155 patients (6.5%). Continued pain was more likely in patients having undergone removal of a normal appendix (P < 0.001) PMID- 7486764 TI - Repeated resection for malignant liver tumours. AB - Fifteen repeat hepatic resections were performed on 12 patients with either recurrent or residual malignant tumours of the liver. Of these, one patient underwent three repeat resections and another underwent two. Five had primary liver liver tumours and seven had liver metastases. Planned, 'staged', repeat resections were performed on three patients because of multiple deposits of tumour, cirrhosis or extensive disease at initial presentation. There was no operative mortality. The period of follow-up from the time of repeat sections ranged between 4 months and 36 months during which two patients died from recurrent disease. The mean survival after the repeat resection was 16.8 months (range 4-36 months). Although technically demanding, repeat hepatectomy is feasible and provide similar benefits. PMID- 7486765 TI - Significant risk factors in elective colorectal surgery. AB - A prospective study of 618 patients undergoing elective colorectal surgery performed in a district general hospital over a 10-year period is reported. Multivariate analysis has been used in an attempt to identify risk factors. The risk of wound infection was increased if septicaemia, respiratory sepsis or faecal contamination was present, if the surgeon was a consultant and if increased risk of serious complications if the patient was male and of poor physical status. Operative mortality was significantly associated with poor physical status, respiratory sepsis and intra-abdominal abscess. Only three risk factors could be identified preoperatively: patient gender, physical status and seniority of surgeon. However, procedures low in the pelvis are more difficult and in this study group are also associated with a higher risk of wound infection. The majority of these procedures are performed by consultants. In addition, this study group has proportionally more female patients of poor physical status who are selectively being operated on by the consultant, seemingly indicating the 'consultant' as a risk factor. PMID- 7486766 TI - Colorectal cancer: surgical prophylaxis and chemoprevention. AB - Attempts at improving outcome from colorectal cancer have recently focused on prevention. This paper describes how both surgical and chemotherapeutic intervention can cause a fall in cell proliferation, polyp regression and a decline in rectal cancer risk in patients with familial adenomatous polyposis. Other patients at high risk of colorectal cancer, or of cancers in other sites, may also have their risks modified by pharmacological intervention. PMID- 7486769 TI - A modification of the pectoralis major myocutaneous flap that reduces the defect at the donor site. AB - A modification of the pectoralis major myocutaneous flap that reduces the cosmetic defect at the donor site is described. PMID- 7486768 TI - The history and evolution of surgical instruments. VI. The surgical blade: from finger nail to ultrasound. AB - Elective surgery requires planned incisions and incisions require appropriate blades. In the prehistoric era, division of the umbilical cord and other minor procedures were probably undertaken with human teeth and nails, and later with plant, animal and mineral substitutes, as witnessed by studies of primitive societies still surviving or recently extinct. More efficient metallic blades appeared in historic times and ultimately generated five specific shapes which are analysed in detail. Today, as minimally invasive techniques, endoscopes, laser and ultrasound sources evolve, many hallowed incisions of surgical access diminish in length or disappear entirely. In historical terms, elective surgery of the twentieth century will be recalled as an interlude characterised by maximally invasive incisions. PMID- 7486767 TI - Early removal of drains and discharge of breast cancer surgery patients: a controlled prospective clinical trial. AB - A prospective trial was conducted to see whether suction drains could safely be removed and patients discharged within 48 h of major breast surgery. Data from two consecutive groups of 50 patients each were compared. Statistical analysis confirmed demographic homogeneity between the two groups with regard to age, tumour size, lymph node involvement, grade of operating surgeon, procedures performed and the 48 h drainage volume. The first group of patients were discharged when drainage was considered acceptable (mean postoperative stay 4.5 days) (long stay). The second group had their drains removed and were discharged after 48 h (short stay). No seromas developed in either group when the total drainage volume (TDV) was less than 150 ml. Seromas developed in 3 (6%) of the long stay group and 5 (10%) of the short stay group (P > 0.05, chi 2 test). No seromas in either group required more than two aspirations. We conclude that it is safe to discharge patients after removal of drains on the 2nd postoperative day. PMID- 7486770 TI - Avoidance of abdominal wall bleeding in laparoscopic surgery. PMID- 7486771 TI - Do-it-yourself femoropopliteal bypass. AB - A 74-year-old male claudicant had undergone operative repair of his bilateral varicose veins 22 years earlier. His records confirmed that on the right side he had only had a saphenofemoral disconnection. Duplex scanning and angiography of his right leg demonstrated an occluded superficial femoral artery (SFA) and a network of channels between the proximal SFA stump and the long saphenous vein (LSV). Distal SFA reconstitution occurred through two distal arteriovenous fistulas. We believe that this is the first case of a spontaneous femorodistal bypass graft secondary to neovascularisation recorded in the literature. PMID- 7486772 TI - Improving exposure and safety at the saphenofemoral junction. PMID- 7486773 TI - Is there a clinical interaction between low molecular weight heparin and non steroidal analgesics after total hip replacement? PMID- 7486774 TI - An audit of surgery of the parotid gland. PMID- 7486775 TI - Management of women presenting to the accident and emergency department with lower abdominal pain. PMID- 7486777 TI - Use of hand-held Doppler to identify difficult forearm veins for cannulation. PMID- 7486776 TI - Use of hand-held Doppler to identify difficult forearm veins for cannulation. PMID- 7486778 TI - Use of laparoscopic instruments in conventional mobilisation of the rectum. PMID- 7486779 TI - Conflicting demands in surgical practice. AB - 975 Fellows of the Association of Surgeons of Great Britain and Ireland were circulated with a questionnaire which was designed to assess the extent to which they experienced conflicts between personal, clinical and administrative demands. Seventy-one per cent of the 305 respondents indicated that they had experienced such conflicts within the previous three weeks. When a conflicts between administrative and clinical demands was experienced, surgeons gave priority to each in similar proportions, but when personal or family needs conflicted with clinical or administrative demands, their personal lives were almost always given a lower priority. The surgeons reported that these types of conflict often affected the quality of patient care and 60 per cent indicated that they were considering early retirement. PMID- 7486780 TI - Improving the primary management of emergency surgical admissions: a controlled trial. AB - The initial screening by senior surgical staff of surgical patients referred for emergency hospital admission should result in improved patient management. The present study was undertaken to determine the effects of this policy. The primary outcome measure was hospital admission rates. The number of operations, diagnostic investigations, initial treatments, deaths, length of stay and bed days per 100 referrals were also measured. The results suggest a 20 per cent reduction in emergency surgical admissions, an important potential benefit to the health service, and to individual patients. PMID- 7486782 TI - Competence assessment of senior house officers in general surgery. PMID- 7486781 TI - Maxillofacial senior house officer posts: the trainee's perspective. AB - A questionnaire was sent out to all the maxillofacial senior house officers in the UK. The purpose of this was to evaluate their perceptions on working conditions, supervision, education and career aspirations. A high proportion (73 percent) returned questionnaires, and from the results of this it seems apparent that there are shortcomings in the supervision and education of SHOs. Based on these findings, a mandatory education programme is advocated that should be recorded in a logbook together with all operative procedures. PMID- 7486783 TI - Operative experience of senior house officers in a district general hospital. PMID- 7486784 TI - Civilian surgeons and the support of the Armed Forces. PMID- 7486785 TI - Off-duty for consultants in the week? It can be done! AB - A work schedule is presented for a single team of three consultant general surgeons which has a rotating three-week timetable with two of them doing routine work only while the other just does the emergencies. It is suggested that this way of working may have widespread application. PMID- 7486787 TI - Surgeons and the new deal--good deal or raw deal? AB - New regulations concerning the training of junior doctors are being introduced. The effect of some of these is already beginning to be noted and the new style Calman training will be introduced shortly. In the South East Thames the implications of Calman type training programme have been piloted in a unified registrar training grade in general surgery for the last three years. It seems an appropriate time to reflect on the changes already operational and the impact of those to come. PMID- 7486786 TI - Shock and dropsy. PMID- 7486788 TI - Inguinal hernia repair: see one, do one, teach one? AB - Clinical management of inguinal hernias has been the subject of recent debate, and guidelines have been produced by The Royal College of Surgeons of England. There is evidence that the results of hernia repair by trainees are inferior to those of more experienced surgeons, but there are no published data on the amount of training and supervision currently experienced by junior surgeons. Thirty-two questionnaires were returned by trainees (21 senior house officers and 11 registrars) from hospitals in the south of England, giving details of teaching and supervision for 3,013 hernia repairs recorded in their log books. Trainees had assisted at 2-90 (median 8) repairs before operating themselves, and had then been supervised during 2-46 (median 9) repairs before being allowed to operate unsupervised. They had assisted subsequently at a further 0-100 (median 9) repairs, and had received supervision during a further 0-42 (median 6) repairs after their first unsupervised repair. The majority (79 per cent) felt adequately trained at the time of their first unsupervised repair. Senior house officers had significantly more experience than registrars in the Shouldice and Lichtenstein techniques, suggesting that these methods have gained ground recently in surgical training. These results show substantial variation in the amount of training and supervision given for inguinal hernia repair. In general, however, trainees experienced a good progression through assisting, supervision, and subsequent reinforcement by further work with senior surgeons after being allowed to operate unsupervised. PMID- 7486789 TI - A day ward in the life of a district urologist: planning the urology day ward list. AB - The day ward activity of a consultant urologist in a district general hospital has been recorded prospectively for the last two years. Particular attention has been paid to the case mix and the time taken. Using this information it has been possible to plan the day ward activity with greater accuracy and efficiency. PMID- 7486790 TI - Can we predict when an operating list will finish? AB - Mean anaesthetic, surgical and turnover times were used to predict finishing times for elective general surgical operating lists. A predicted early finish was correct in 70 per cent, a predicted on-time finish in 19 per cent, and a predicted late finish in 56 per cent. Overall, predictions of an early or late finish had a low sensitivity (62 per cent and 65 per cent) and high false positive rate (30 per cent and 44 per cent). Over-runs, caused by too many cases, and early finishes, owing to insufficient cases were reliably predicted. It is suggested that mean procedure times can be used to identify under or over utilisation caused by under or over-booking but indiscriminate use will not improve list utilisation. PMID- 7486791 TI - Formal assessment of the surgical trainee: a review of eight years experience at St Lawrence Hospital, Chepstow. AB - In light of the proposals for a continuum of higher surgical training, we reviewed an 8-year experience of formal assessments of surgical trainees. We undertook a retrospective analysis of 34 completed assessment forms, retrieved from records, and a postal questionnaire of 54 former (and current) trainees. Our study revealed that all trainees agreed in principle with formal assessments. Eighty-one per cent found their assessment either useful or excellent and a further 17 per cent found the advice received was influential in their careers. The consultants, in turn, were able to modify the training programme as a result of constructive feedback. Based on our experience we recommend formal assessments of surgical trainees as an important and rewarding part of an impartial peer review system within a continuum of higher surgical training. PMID- 7486793 TI - The determination of bio-available testosterone. PMID- 7486792 TI - James Moore, John Hunter and amputation under analgesia in 1784. PMID- 7486794 TI - Factors affecting the measurement of cytokines in biological fluids: implications for their clinical measurement. PMID- 7486795 TI - Wide diversity in measurements of growth hormone after stimulation tests in short children are due to assay variability. AB - A comparison of three commercially available assays for growth hormone (Pharmacia hGH RIA, IDS Gamma-BCT GH and Delfia 22 kDa hGH) were used to measure growth hormone (GH) secretion in 26 short children after pharmacological stimulation. The IDS Gamma-BCT assay was compared with Pharmacia assay in 15 provocative tests (13 children; n = 94). The Pharmacia assay showed a strong positive proportional bias and this was particularly marked at GH > 20 mU/L; the mean difference between the assays at GH < 10 mU/L was 12%. The IDS Gamma-BCT assay was also compared with the Delfia 22 kDa assay in 18 provocative tests (13 children; n = 100). Results from the IDS assay was higher throughout the range 0-90 mU/L with a mean difference of 11.4% in the range 10-90 mU/L; the mean difference between the assays at GH < 10 mU/L was -39%. PMID- 7486796 TI - Choice of time for urine collection for detecting early kidney abnormalities in hypertensives. AB - The aim of this study was to determine the most appropriate urine collection for detecting differences in the excretion rates of albumin, gamma glutamyl transpeptidase (GGT) and N-acetyl-beta-D-glucosaminidase (NAGA) between normotensive subjects and hypertensive patients on treatment. Twenty treated hypertensive patients, mean (SEM, standard error of mean) age; 52.2 (6.2) years and 20 normotensive subjects, mean age 49.2 (4.2) years, were studied in a consecutive sampling design. Urinary excretion rates of albumin, GGT and NAGA were determined in consecutive timed urine samples collected overnight and during 3-5 h the next morning. Mean (SEM) overnight excretion rates for albumin, GGT and NAGA for normotensive subjects were 11.05 (1.18) micrograms/min, 17.00 (2.20) mU/min and 6.55 (0.39) mU/min, respectively, which were significantly lower than those of hypertensive subjects which were 20.77 (2.14) micrograms/min, 21.84 (1.65) mU/min and 10.92 (0.87) mU/min, respectively (P < 0.05). The mean (SEM) percentage increases in urinary albumin, GGT and NAGA in morning urine collections of normotensive subjects of 15.22 (3.88)%, 34.04 (6.45)% and 11.54 (3.63)%, respectively were significantly lower than 107.03 (15.04)%, 121.96 (16.71)% and 72.75 (7.50)% found in hypertensive patients (P < 0.05). These data suggest that were urinary albumin and tubular enzyme excretion to be used as correlates of hypertensive renal damage, ambulatory urine collections may be more sensitive than overnight collections. PMID- 7486798 TI - Euthyroid sick syndrome in pulmonary tuberculosis before and after treatment. AB - Alterations of circulating thyroid hormones are frequently present in chronic nonthyroidal illnesses and may predict prognosis. Pulmonary tuberculosis, a common treatable debilitating disease, may provide a useful model for detailed evaluation of changes of thyroid hormones in relation to subsequent recovery or mortality. Over a period of 12 months, we performed a prospective study of 40 consecutive Chinese patients aged over 50 years and admitted with newly diagnosed pulmonary tuberculosis. Blood samples were drawn for serial thyroid function tests [free thyroxine (T4), free triiodothyronine (T3) and thyroid-stimulating hormone] before treatment and at 1, 2 and 4 months afterwards. Mortality was determined up to 12 months of follow-up. The euthyroid sick syndrome occurred in 63% of patients at presentation. Twelve of 25 euthyroid sick patients died as compared to one of 15 patients with normal baseline thyroid function tests (P < 0.02). Among euthyroid sick patients, those who died had significantly lower free T3 concentration at presentation than those who survived (P < 0.05). An undetectable free T3 concentration at presentation was associated with a subsequent mortality of 75% (9 of 12). Of the survivors, all patients demonstrated a significant rise in serum free T4 concentrations following treatment, which was apparent by 1 month. These data suggest that an undetectable free T3 concentration at presentation reflects severity of illness and predicts a subsequent high mortality. PMID- 7486797 TI - Bone alkaline phosphatase in rheumatic diseases. AB - A double monoclonal immunoradiometric assay specific for bone alkaline phosphatase (BAP) was used to determine whether the raised total alkaline phosphatase (TAP) often found in patients with active rheumatoid arthritis (RA) and ankylosing spondylitis (AS) is derived from bone or liver. Fifty-eight patients with RA were compared to 14 with AS and 14 with non-inflammatory rheumatic diseases (NI). None had clinical liver disease and only one had a slightly elevated aspartate transaminase activity. Elevated BAP concentrations were found in seven patients (5 RA, 1 AS, 1 NI), only two of whom also had abnormal TAP. Abnormal TAP activities were found in only three patients (all RA). BAP did not correlate with disease activity in RA or AS. In contrast, TAP correlated with disease activity (assessed by plasma viscosity) in RA (P < 0.002) and gamma-glutamyl transferase (GGT) also correlated with plasma viscosity in RA (P < 0.01). Both TAP and BAP were significantly correlated with GGT in RA (P < 0.001 and P < 0.02, respectively). These findings are discussed, together with possible reasons for the conflicting nature of some of the observations. PMID- 7486799 TI - Proton nuclear magnetic resonance lipid profiling of intact platelet membranes. AB - A combination of high-resolution one-dimensional and two-dimensional proton nuclear magnetic resonance spectroscopic methods and high pressure liquid chromatography have been used to analyse qualitatively and quantitatively the phospholipids, cholesterol and fatty acid composition of normal human blood platelets. The major phospholipids present were identified and quantified. Information on the nature and composition of the different classes of structural phospholipids and unsaturated fatty acids was obtained and was similar to that reported by others using different analytical procedures. This work showed that the proton nuclear magnetic resonance spectroscopy approach to lipid analysis could provide rapid, comprehensive and detailed information on the roles of lipids in pathological conditions in which platelets have been implicated. PMID- 7486800 TI - Induction of M-CSF receptor and its mRNA, and activation of tyrosine kinase in peripheral monocytes by oestradiol-17 beta and progesterone. AB - In monocytes at the secretory (oestrogen-progesterone dominant) phase of the menstrual cycle, expression of c-fms and macrophage colony-stimulating factor (M CSF) receptor and activity of tyrosine kinase (TK) were increased by oestradiol with or without progesterone. In vivo, oestrogen may induce expression of c-fms and M-CSF receptor as well as the activation of TK in monocytes under the milieu of the secretory phase. Alternatively, cells of monocyte lineage during the secretory phase might, via various factors, obtain the potency to induce the expression and the function of M-CSF receptors, this potency being effected by oestrogen. Macrophages in peritoneal fluid in pelvic endometriosis (oestrogen predominant) might be activated during the secretory phase of the menstrual cycle, causing infertility. PMID- 7486801 TI - Mass versus activity: validation of an immunometric assay for bone alkaline phosphatase in serum. AB - A detailed investigation of the performance of an immunometric assay for the quantitation of bone alkaline phosphatase (ALP) in serum showed that precision of this assay was similar to that of lectin precipitation and electrophoretic methods. Relationships for mass and activity measurements of the bone isoform using samples from children and patients with Paget's disease were similar for lectin and electrophoretic methods. Purified liver enzyme showed 100% cross reactivity in the immunometric assay. However, comparison of the slopes between bone ALP mass and total ALP activity using only samples with predominantly liver or bone isoforms showed that the cross reactivity of the liver isoform in serum was 18.3%. Experiments in which increasing amounts of a sample containing 90% of liver isoform adding to a serum sample from a patient with Paget's disease showed a cross reactivity of 16.5%. The reference range for bone ALP mass was 7-28 micrograms/L for men (n = 77) and 5-20 micrograms/L for women (n = 110) in the age group 20-50 years. In women over 50 years bone ALP was 28% higher. Increased bone ALP mass was also demonstrated in patients with Paget's disease (n = 59), liver disease (n = 95), chronic renal failure (n = 41) and hyperthyroidism (n = 17). PMID- 7486803 TI - Urinary iron excretion in patients with chronic obstructive airways disease (COAD). PMID- 7486802 TI - The effect of anticoagulant choice on apparent total antioxidant capacity using three different methods. AB - We assessed total antioxidant capacity using three different methods, in plasma samples treated with either EDTA or heparin as anticoagulant, from 26 healthy subjects. Total antioxidant capacity was determined using an oxygen electrode (as the total peroxyl radical-trapping antioxidant parameter), by enhanced chemiluminescence, and by measurement of the antioxidant-mediated quenching of the absorbance of a radical cation. The choice of anticoagulant had a profound effect on antioxidant capacity with heparinized plasma giving consistently higher values than plasma anticoagulated with EDTA. Using the oxygen electrode the mean value was 786.5 +/- 171.5 mumol/L (heparin) compared to 681.4 +/- 160.4 mumol/L (EDTA, P < 0.01). The chemiluminescence technique gave a mean antioxidant capacity of 915.6 +/- 214.1 mumol/L in heparin samples and 714.4 +/- 195.4 mumol/L in EDTA samples (P < 0.0001). The absorbance quenching technique gave a mean value of 867.0 +/- 199.2 mumol/L (heparin) and 675.5 +/- 245.4 mumol/L (EDTA, P < 0.001). All methods tested showed comparable results for EDTA plasma, but the chemiluminescence technique gave higher apparent antioxidant capacity than either of the two techniques when heparin plasma was used. We suggest that either heparin is interacting to enhance antioxidant protection perhaps through release of superoxide dismutase, or the chelation of metal ions by EDTA is limiting the activity of antioxidant metalloenzymes. Consistency in the choice of anticoagulant is clearly extremely important. PMID- 7486804 TI - Contribution of serum globulins to total fructosamine in patients treated by haemodialysis. PMID- 7486805 TI - Differences in the stability of intact osteocalcin in serum, lithium heparin plasma and EDTA plasma. PMID- 7486806 TI - Positive interference by azapropazone (Rheumox) in the measurement of total thyroxine using the Ciba-Corning ACS:180 immunoassay analyser. PMID- 7486807 TI - Renin-angiotensin-aldosterone and kallikrein investigations in a patient with resistant hypomagnesaemia due to Gitelman's syndrome. PMID- 7486808 TI - Mivacurium sensitivity in a patient heterozygous for the atypical and silent genes for plasma cholinesterase. PMID- 7486809 TI - Classification of an alloalbumin. PMID- 7486810 TI - Mononuclear blood cell magnesium in older subjects: evaluation of its use in clinical practice. PMID- 7486811 TI - Implications of human T-lymphotropic virus type-I and type-II testing in donors and patients. AB - The retroviruses known as Human T-Lymphotropic Virus Types I and II (HTLV-I and II) were recognized before the human immunodeficiency virus (HIV-1). Associated diseases of HTLV-I infection, including a particular kind of leukemia or the development of a specific demyelinating disease, have also been observed. Screening of blood donors for antibodies to HTLV was mandated in November of 1988. This paper examines the biology of HTLV-I and HTLV-II and reviews the testing methods for HTLV-I/II. Data from 39,908 blood donations of volunteer donors at The University of Texas M. D. Anderson Cancer Center (UTMDACC), Division of Laboratory Medicine, Section of Transfusion Medicine are presented. Initially reactive specimens for HTLV antibodies were 158 (0.4 percent). Of these 0.26 percent or 105 of 39,908 were repeatedly reactive. Eight hundred and sixty seven cancer patients were also tested for HTLV antibodies. Eight or 0.9 percent were repeatedly reactive for HTLV antibodies by enzyme immunoassays (EIA), but only one could be confirmed as positive. HTLV-I/II has a very low incidence in the ambulatory population. The relationship of clinical sequelae and the rate of transmission of these viruses remain unclear. A readily applicable confirmatory test is not yet available. Even significant improvements in the sensitivity and specificity of testing will present ongoing problems for identification of true HTLV carriers. The clinical decision-making process related to the meaning of these results continues to be difficult. PMID- 7486814 TI - Intracellular calcium and hydrogen ions in diabetes mellitus. AB - Diabetes mellitus is a multi-component syndrome that is often complicated by angiopathy which is partly due to enhanced platelet functions. Using fluorescent dyes 2',7'-bis(carboxyethyl)-5(6)-carboxyfluorescein (BCECF) and Fura-2 AM, changes was evaluated in the concentration of baseline and thrombin-stimulated increases in intracellular ionized calcium (Ca2+i) relative to hydrogen ions in the platelets from control, insulin-treated, and non-treated diabetic rats. The cytosol of platelets from the diabetic rats were more acidic compared to the insulin-treated and normal control rats. The increased intracellular hydrogen ion concentration [H+] or decreased pH (pH) in the diabetic rat platelets is associated with an increased baseline [Ca2+]i. Upon stimulation with thrombin, the mean peak [Ca2+]i for the insulin-treated (309 +/- 97 nmol/L) and untreated (339 +/- 135 nmol/L) diabetic rats was significantly higher than the concentration for the normal rats (213 +/- 101 nmol/L). Treatment with insulin attempts to correct the diabetes-induced elevation in the baseline of [Ca2+]i and intracellular H+. These results suggest that the relationships between Ca2+ and H+ relative to binding sites are similar in the intra- and extracellular compartments. It is our conclusion that the enhanced platelet activity and associated development of vascular diseases in diabetes may be due to an increased intracellular H+ that caused an increased baseline [Ca2+]i in diabetes mellitus. PMID- 7486812 TI - Testosterone increases insulin-like growth factor-1 and insulin-like growth factor-binding protein. AB - Growth of the male external genitalia is primarily regulated by androgens; however, several observations suggest growth hormone (GH) or a GH dependent factor, such as insulin-like growth factor-1 (IGF-1), might also be involved. It is hypothesized that testosterone (T) might induce the synthesis of IGF-1 or IGF binding protein (IGF-BP) which could affect cell proliferation. This was evaluated by determining the effect of T on thymidine incorporation, cell surface IGF-1 binding, and the production of IGF-1 and IGF-BP by cultured neonatal foreskin fibroblasts. Testosterone significantly increased thymidine incorporation and the production of IGF-1 and IGF-BP (p < 0.05 vs control). However, T significantly decreased the cell surface binding of IGF-1 (p < 0.0001 vs control). To determine whether or not the increase in IGF-1 production was important in mediating the effect of T on thymidine incorporation, cells were incubated with either anti-IGF-1 antibody (anti-IGF-1), anti-IGF-1-receptor antibody (IGF-1-R-Ab), or a non-specific control antibody (NS-Ab). Anti-IGF-1 significantly decreased thymidine incorporation in both control cultures and those containing T. In addition, IGF-1-R-Ab blocked the expected T dependent increase in thymidine incorporation, while NS-Ab had no effect. These in vitro observations suggest both T and IGF-1 affect neonatal foreskin fibroblasts in a complex relationship. In addition, these data suggest T might stimulate foreskin fibroblast proliferation, at least in part, by changing the balance in production and effects of IGF-1 and IGF-BP. PMID- 7486813 TI - Cytogenetics, gene fusions, and cancer. AB - Cytogeneticists recognize that karyotypic abnormalities are associated with specific malignancies. In 1960, Nowell described the Philadelphia chromosome (Ph) and its relationship to chronic myelogenous leukemia (CML). Subsequent work in molecular genetics and biology has revealed that the Ph is a translocation that causes fusion of gene sites that code for the break cluster region (BCR) and the avian blastic leukemia (ABL) proteins. This so-called fusion protein is present in a large percentage of the patients who have CML. A related fusion protein is seen in about one third of patients with acute lymphoblastic leukemia. The BCR ABL fusion protein results in increased tyrosine kinase activity. The mechanism of action is thought to be via signal transduction related to guanosine triphosphatase activating protein which interacts with a ras-p21 binding protein. Acute promyelocytic leukemia (APL) is associated with the cytogenetic abnormality of t(15;17). This alters the promyelocytic leukemia (PML) and the retinoic acid receptor alpha (RARA) gene sites. Two fusion proteins are the result of this cytogenetic abnormality. They are termed PML-RARA and RARA-PML. Only one, the PML RARA, is associated with APL. The PML-RARA chimeric protein has two zinc finger like regions. It retains the ligand binding domain of RARA. The protein called PML has some similarities with a family of proteins which are thought to fuse to proto-oncogenes and to act as transforming proteins. The role of classical cytogenetics and the added capability of molecular biology has helped to elucidate some of the potential mechanisms for the development of cancer and provided additional understanding of neoplasia. (ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7486815 TI - Assessment of sex chromosome composition using fluorescent in situ hybridization as an adjunct to GTG-banding. AB - Fluorescent in situ hybridization (FISH) using dual color X chromosome- and Y chromosome-specific probes was employed to assess further the sex chromosome copy number in cells of a phenotypic female patient with hypergonadotropic hypogonadism, primary amenorrhea and growth retardation. The GTG-banding analysis of peripheral blood lymphocytes had revealed the presence of predominantly 46,XY cells. A FISH analysis, undertaken to assess further the contribution of a minor cell line, yielded frequencies of 87% cells with the 46,XY constitution and 9% with the 45,X constitution. To establish unequivocally the presence of mosaicism, a skin biopsy was obtained for fibroblast culture, which further corroborated the results of the peripheral blood study. Fluorescent in situ hybridization analysis revealed 74% of the cells to be 46,XY and 12% to be 45,X. The unequivocal presence of XY cells puts the patient at risk for neoplastic transformation of the gonads. Laparoscopy and surgical removal of the patient's presumptive streak gonads were therefore undertaken. Cytogenetic results derived from the gonadal tissues further strengthened findings of previous cytogenetic analyses. It is our experience that FISH is a useful adjunct to established cytogenetic techniques in the management and monitoring of patients similar to the proband described in this study. PMID- 7486816 TI - Granulomatous hepatitis as a complication of intravesical Bacillus Calmette Guerin therapy for bladder carcinoma. AB - Bacillus Calmette-Guerin (BCG) therapy has been proven to be effective treatment for superficial bladder carcinoma with complications that are usually minor and self limited. Although rare, granulomatous hepatitis can develop, and it is important to recognize it and treat it promptly. The hepatitis which occurs appears to be the result of a hypersensitivity reaction to antigens present in the BCG vaccine. Pathological findings include hepatocellular necrosis and pleomorphism, microgranulomas and non-caseous epitheliod granulomas with giant cells. A case of BCG-hepatitis, which developed in a 74-year-old white male following immunotherapy for superficial bladder carcinoma, is described. PMID- 7486818 TI - [Care patterns and resumption of social and occupational activities after exeresis surgery in Crohn's disease. Study of 58 patients]. AB - The aim of this retrospective study was to describe care patterns and conditions surrounding the resumption of social and work activities after intestinal resection for Crohn's disease. Fifty-eight patients were evaluated regarding their stay in the department of surgery, use of care and resumption of social and work activities within the first year after intestinal resection. Data were obtained from medical records and answers to a medical questionnaire. Forty patients replied to the questionnaire. Mean length of stay in the department of surgery was 16 +/- 9 days. Length of stay was positively correlated to non elective surgery, colonic resection, presence of fistulae or abscesses, and to the creation of a stoma. Main interval between surgery and resumption of work was 11 +/- 8 weeks. Only one patient was unable to return to work because of the disease. Seventy per cent of the patients who went back to work said that their quality of life was improved after surgery in comparison with their preoperative status. In conclusion, this study suggests that most of the patients undergoing intestinal resection for Crohn's disease can go back to work without particular difficulties, and consider that their quality of life has been improved by surgery. PMID- 7486817 TI - [Anorectal malignant melanomas. Apropos of 5 cases. Review of the literature]. AB - This study reports five cases of anorectal melanoma treated at Centre Val d'Aurelle-Paul Lamarque in Montpellier. There follows a discussion based upon the literature of epidemiological, clinical, histopathological and therapeutic aspects. Surgery is the treatment of choice but there is major controversy regarding the best method. The decisive prognostic factor appears to be the stage of the disease rather than the type of treatment. The role of radiotherapy, adjuvant chemotherapy and immunotherapy remains to be evaluated. PMID- 7486819 TI - [Tumor seeding of the abdominal wall after fine needle cytologic puncture of the liver. Apropos of a case]. AB - Tumor seeding of the abdominal wall during fine needle puncture-aspiration of the liver is a very rare complication. The authors report a case of this complication affecting a 50-year-old Senegalese woman. She underwent echo-guided fine needle cyto-puncture of a hepatocellular carcinoma arising from the left lobe, followed by left lobectomy. The subcutaneous nodule was discovered 42 months after cyto puncture. Histology following its excision revealed infiltration of carcinomatous hepatocytes between the fibers of rectus abdominis. There was no recurrence and the patient was well 10 months after excision of the subcutaneous nodule. With a review of 10 others cases reported in the literature, the authors feel that, although rare, this complication must be avoided and suggest a system for the protection of structures through which the needle passes. PMID- 7486821 TI - [Inflammatory bowel diseases in children]. PMID- 7486820 TI - [Choice of treatment in rectal cancer. Consensus Conference, Chamber of Commerce and Industry, Paris, December 1-2, 1994]. PMID- 7486822 TI - [Food-induced anaphylaxis. A new French multicenter survey]. AB - The frequency of food-induced anaphylactic shocks (FIAS) has been investigated beside 46 emergency departments, 29 departments of Dermatology, and 19 departments of Intern Medicine or Pneumology. 794 A.S. have been reported. FIAS represent 10.2% of the etiologies. More than a third of them are relapsing anaphylaxis. Food allergy had been previously identified in 23.4% of cases. The allergen was present as a hidden allergen or inadvertently consumed in 30.8% of FIAS. 9.9% of the patients were asthmatic. An enhancing factor was heightened in 25.9% of cases: alcohol, exercise, simultaneous intake of aspirin, beta-blockers, conversion enzyme inhibitors. Other factors predisposing to food anaphylaxis were a cross-reactivity shared by pollens and fruit, latex and exotic fruit, house dust mites and snails, or mastocytosis. More than 15 allergens were detected: egg (11.6%), fish (10.4%), crustaceans (10.4%), milk (6.5%), fruit-latex group (6.5%), peanut and other leguminosae (soy, peas, lentils, guar gum...), celery, garlic, etc... The food allergen still remained unknown in 25% of cases. However, the rate of efficiency of the diagnosis reached 94% in the Allergy Units. In addition 10.2% of A.S. were considered idiopathic, raising the hypothesis of allergy to masked food allergens. Compared to a previous study from the 1982's, this survey shows a striking increased prevalence of FIAS.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7486823 TI - [Comparison of the effects of Rocgel and anti-H2 on the symptomatology of gastroesophageal reflux without esophagitis]. AB - The efficacy of boehmite (Rocgel) and of ranitidine was compared in a randomized, double-blind 4-week trial in patients with symptomatic, endoscopically free macroscopic reflux oesophagitis. Of the 64 patients who completed the study 33 received boehmite and 31 ranitidine. Significant symptomatic improvement occurred in both treatment groups (global clinical score and self evaluation by patients) (p < 0.001). Disappearance of heartburn was 52 per cent (15/29) with boehmite and 53 per cent (16/30) with ranitidine. Disappearance of regurgitation was 48 per cent (10/21) with both treatments. 33% of the patients became totally symptom free. Our results indicate that boehmite seems to be at least as effective as ranitidine in relieving symptoms. Cost of treatment with boehmite, on the other hand is cheaper than ranitidine. As a safe, locally active mucosal protecting agent and antacid, boehmite is an effective drug for the treatment of reflux oesophagitis without macroscopic lesions. PMID- 7486824 TI - Detection of t(X;Y) in 2 XX males using fluorescent in situ hybridization. AB - Males with a 46,XX karyotype generally have Y chromosomal material translocated to the pseudoautosomal region of the X chromosome. We have delineated two such cases using two color fluorescent in situ hybridization with probes from the short arm (DYZ2), centromere (DYZ3), and long arm (DYZ1) of the Y chromosome and a centromeric probes for the X chromosome (DXZ1). Using these techniques, the two patients are identified as having the karyotype 46,X,der(X)t(X;Y) (p22;p11) and a phenotype consistant with this translocation. Azoospermia in these patients is explained by the absence of Y long arm material including the recently identified candidate gene family for spermatogenesis. PMID- 7486825 TI - Early urethral obstruction sequence and trisomy of the long arm of chromosome 1. PMID- 7486826 TI - Identification of a polymorphism in the coding region of the p27Kip1 gene. PMID- 7486827 TI - DiGeorge syndrome and related syndromes associated with 22q11.2 deletions. A review. AB - DiGeorge syndrome (DGS) is a developmental defect which associates hypo- or aplasia of the thymus and parathyroids, facial dysmorphism and conotruncal cardiac malformations. The etiological factor in a great majority of DGS patients is monosomy for the 22q11.2 chromosomal region either through a large interstitial deletion of that region (inherited or de novo) or through an unbalanced translocation involving chromosome 22. In one instance, a balanced translocation of chromosome 22 was associated with a DGS phenotype. Extensive analyses of this region of chromosome 22 has led to the obtention of precise physical maps of the corresponding genomic region, to the cloning of the balanced translocation breakpoint and to the isolation of different genes from the minimal critical deleted region. PMID- 7486828 TI - [Centenary of the birth of Raymond Turpin (1895-1988). A reminiscence of his life and work. Exerpts from the address of Professor Robert Laplane to the Academie de Medecine, 9 May 1989]. PMID- 7486830 TI - [Direct analysis of the frequency of disomy in human sperm using the PRINS technique]. AB - The PRINS method allows a rapid and specific detection of human chromosomes in situ. We have adapted the PRINS protocol to human sperm. Estimates of disomy have thus been performed for chromosomes 9, 12, 16 and 21 by using alpha-satellite DNA specific primers. The frequencies of disomy ranged from 0.27% to 0.31%. No significant difference was found. These data agree with the hypothesis of an equal distribution of non-disjunctions among chromosomes in male meiosis. PMID- 7486831 TI - [Demonstration of transposition in Portulaca grandiflora (HOOK)]. AB - The gene C of Portulaca grandiflora encodes a product required for the synthesis of flower pigment. The cmI allele gives white flowers with violet spots and sectors. It is also germinally unstable and can give rise to a series of cm alleles conferring different flower color phenotypes. These findings suggest that allele C may be rendered genetically unstable as a consequence of transposable element insertion and excision. PMID- 7486832 TI - Autosomal recessive omodysplasia. AB - The authors report on a new case of omodysplasia with severe shortness of lower limbs and with first cousin parents. This condition is a second form of omodysplasia. Recessive transmission is confirmed by consanguinity. PMID- 7486829 TI - Chromosomal localization of two KOX zinc finger genes on chromosome bands 7q21 q22. AB - Human cDNAs encoding Kruppel-type zinc finger domains, designated KOX 1-32, have been cloned from human T lymphocyte cell line libraries. We report here the regional localizations by in situ hybridization of KOX 18 and KOX 25 on chromosome bands 7q21q22. Pulse-field gel electrophoresis (PFGE) analysis showed that these genes are physically located within a DNA fragment of 250 kb. The genes KOX 4 and KOX 9, mapped on chromosome 8q24, were found to be located within a DNA fragment of 450 kb. From the present and previous data, eighteen different KOX genes have been located at least two by two within nine DNA fragments of 200 to 580 kb. PMID- 7486833 TI - Genetic variation in the COL6A1 region is associated with congenital heart defects in trisomy 21 (Down's syndrome). AB - Genetic variation in the COL6A1-COL6A2 gene cluster on chromosome 21 was studied in 113 controls and 58 European families (including control and family subgroups of British/Irish origin) having a child with trisomy 21. There were statistically significant differences among subgroups of trisomic children with and without congenital heart defects (CHD) in distributions of definitive, 3-RFLP haplotype classes received from their nondisjoining and disjoining parents. Haplotypes received by trisomic children with CHD from their disjoining parents were not a random sample of controls' haplotypes. Analysis of parental single-RFLP genotypes and linkage disequilibrium patterns confirmed this parent subgroup differed from a random sample of controls. There were no significant differences in parent subgroup genotype distribution at any of nine control loci distributed along chromosome 21q. This sample showed an association between genetic variation in the COL6A1 gene region and congenital heart defects in trisomy 21. PMID- 7486834 TI - Genetic epidemiology of breast cancer: interest of survival analysis methods. AB - In an attempt to explain the controversy resulting from the analysis of the breast cancer data collected by Jacobsen, a segregation analysis was performed using successively the unified mixed model (UM) and the logistic hazard function model (LHM) (Abel & Bonney, 1990). Under the UM, age of onset of the disease cannot be taken into account, each individual being assigned to a liability class according to his age at examination, whereas, in the LHM, variable age of onset is modelled using survival analysis methods. Under the UM, we confirmed the results of Demenais et al. (1986b), i.e. the transmission probabilities are significantly different from Mendelian expectations. The same results were obtained when taking into account the specific mortality for the computation of the morbid risk observed in a given liability class. Under the LHM, the analysis provides evidence for a monogenic autosomal model with a rare dominant allele responsible for the disease, and transmission probabilities compatible with Mendelian expectations. This study shows that the rejection of the Mendelian transmission under the UM can be due to a violation of a constraint of this model (i.e. the probability of being and not being affected in a given liability class should sum to 1) when a specific mortality is induced by the disease as in breast cancer. Survival analysis methods avoid these problems by taking into account the onset of the disease as the failure time event and are more suitable when studying a complex trait such as breast cancer. PMID- 7486835 TI - Evolutionary dynamics of the FMR1 locus. AB - Rare haplotypes for close flanking markers are associated with increased allele size and frequency of the fragile X mutation. Exceptional founder haplotypes can be identified, but many haplotypes with rare alleles contribute to full mutations. A transition matrix constructed from the data predicts that a population with reduced variability will manifest a slowly increasing frequency of premutations and full mutations, reach a distribution close to the observed one after a few hundred generations, and then slowly be depleted of these alleles. This prediction is opposite to less well supported inference of increasing frequency of progressive amplification, but the data are inadequate to reach any firm conclusions. Factors that may determine the evolution of these systems, but cannot now be evaluated, are discussed. PMID- 7486836 TI - Integration of gene maps: updating chromosome 1. AB - The first integrated map of chromosome 1 was published in 1992. We present an updated summary map of 371 loci constructed from a location database that includes physical and genetic data. The summary map subsumes a composite physical location, sex-specific genetic location, cytogenetic assignment, mouse homology, rank and references to physical maps. The genetic length is 208 cM for the male map, in close agreement with the chiasma map, and 371 cM for the female map. There is evidence for a high level of interference on chromosome 1. The location database comprising both data and analytical software is discussed in relation to alternative approaches and possible enhancements. PMID- 7486837 TI - Estimating multipoint recombination fractions. AB - The likelihood in linkage investigations can be expressed as a polynomial in the relevant recombination fractions. With polymorphic characters, this can be reduced to a product of separate factors for female and male recombinations, with only mild loss of information. With three generation data the polynomials take a specially simple form. Algorithms are given for finding maximum likelihood estimates and their standard errors. PMID- 7486838 TI - An extended transmission/disequilibrium test (TDT) for multi-allele marker loci. AB - The transmission/disequilibrium test (TDT) was recently introduced by Spielman et al. (1993) as a test for linkage and linkage disequilibrium. The test is based on the unequal probability of transmission of two different marker alleles from parents to affected offspring, when the marker locus and the hypothetical disease locus are linked and are in linkage disequilibrium. The probabilities of marker allele transmission to affected offspring conditional on parental genotype have been derived by Ott (1989) for a biallelic marker and a recessive disorder with no phenocopies. Here, we derive the transmission probabilities for a multi-allele marker locus and a generalized single locus disease model in a random sample of affected individuals from a randomly mating population. The form of these transmission probabilities suggests an extension of the TDT to multi-allele marker loci, in which the alternative hypothesis is restricted to take account of the likely pattern of unequal transmission when the recombination fraction is near 0. We show how our extended TDT can be implemented by standard software for logistic regression, although we have also written our own program which is available on request. We have evaluated the approximate power of the test under a range of realistic assumptions, and it appears that the test will often have good power when linkage disequilibrium is strong and if the disease is recessive. PMID- 7486839 TI - Comparison of changes in energy expenditure and body temperatures after caffeine consumption. AB - Twelve young healthy male volunteers participated in a randomized blind study in which they consumed 150 ml decaffeinated coffee with or without 200 mg added caffeine. Resting metabolic rate by indirect calorimetry (ventilated hood), skin temperature on chest, arm, thigh, calf and abdomen, and internal temperature (rectal) were measured in the fasting state and up to 3 h after coffee consumption. All treatments were done in duplicate. After caffeine consumption the metabolic rate increased immediately with 0.2 +/- 0.2 kJ/min (p < 0.05) and remained elevated for the 3 h during which measurements were taken. Mean total caffeine-induced thermogenesis was 0.30 +/- 0.20 kJ/min, which means a mean increase in the metabolic rate of 7 +/- 4% during 3 h. The internal temperature started to increase after both treatments, but became slightly different 2 h after caffeine consumption. Mean skin temperature after caffeine consumption increased, and became significantly different compared to the control treatment after 90 min. Thus, the increase in both internal temperature and skin temperature after caffeine consumption was largely delayed compared to the direct detectable increased energy expenditure as measured by indirect calorimetry. The correlation between the change in metabolic rate during the 3 h after the caffeine consumption and the mean change in skin temperature was low (r = 0.38; p = 0.06), and does not seem to justify the use of skin temperature to measure energy expenditure or changes in energy expenditure in short-term studies. PMID- 7486840 TI - Differential incorporation of 13C label from dietary glucose into neutral sugars of rat intestine macromolecules. AB - Using gas chromatography-isotope ratio mass spectrometry, the detection, with a good reproducibility and in a single step, of the 13C content of glycoprotein neutral sugars, glycogen glucose, and ribonucleic acid ribose allowed us to trace incorporation into intestinal macromolecules of sugars derived from dietary glucose, labeled either naturally (as corn starch) or artificially. The 13C enrichment of glycoprotein neutral sugars was strong and rapid and plateaued up to the end of the experiment, whereas the 13C content of ribose increased linearly with time. By contrast, with artificially enriched dietary glucose, no significant 13C enrichment was detected in glycogen, suggesting that this macromolecule does not directly derive from dietary glucose. These results suggest differential control of sugars involved in the synthesis of intestinal macromolecules and demonstrate that this technique would be sensitive and reliable enough to be applied in man for serum glycoprotein sugar determinations. PMID- 7486841 TI - Effect of chronic ethanol administration and dietary protein regimens on intestinal absorption of macromolecules in rats. AB - The effect of feeding ethanol daily for 40 days has been studied on intestinal absorption of bovine serum albumin (BSA) and gamma-globulin (IgG) in rats fed a low (8%) protein (LP) or a high (30%) protein (HP) diet. Feeding the LP diet enhanced the tissue uptake of BSA (p < 0.05) and absorption of BSA and IgG into serum (p < 0.001) as compared with controls. Feeding the HP diet also augmented the uptake of IgG (p < 0.001) by the intestinal tissue and significantly enhanced serum levels of BSA and IgG. Ethanol feeding to rats for 40 days enhanced the uptake of BSA and IgG (24-84%) and their absorption into serum (p < 0.001) as compared with the controls. Ethanol administration to rats fed LP or HP diets did not alter the uptake of these proteins as compared with their respective controls. Luminal degradation of BSA and IgG was higher in ethanol-administered (48-50% and 36-39%, respectively) and LP-fed rats (50 and 38%, respectively). It was reduced by 17-21% in HP-fed rats as compared with the control group. This indicated that the observed increase in protein absorption is not related to the luminal degradation of the proteins under these conditions. These findings suggest that the absorption of macromolecules from intestine in response to ethanol feeding is influenced by the dietary status of the animals. PMID- 7486842 TI - Comparison of fluoxetine and placebo in the treatment of obesity. AB - Our study aims at assessing the efficacy and safety of fluoxetine as compared with placebo in the treatment of obesity using a double-blind crossover design. We studied 42 obese women (body mass index 35.9 +/- 5.3 kg/m2). The obese patients were randomized to start treatment with fluoxetine (group A) or placebo (group B) for 3 months (period 1). After a 1-month washout period, treatment was crossed for the following 3 months (period 2). There was no significant difference in weight loss when the patients were treated with fluoxetine (group A period 1 and group B period 2) as compared with patients treated with placebo (group B period 1 and group A period 2). There were no significant differences in monthly weight reduction during both treatments. In conclusion, we demonstrated that serotoninergic drugs such as fluoxetine need further investigation before being used indiscriminately in obese subjects. PMID- 7486844 TI - Determination of intracellular water by multifrequency bioelectrical impedance. AB - Body composition was measured in 57 healthy males by 40K measurements and by multifrequency bioelectrical impedance. Intracellular resistance (Ricf) was calculated from the impedance values using the Cole-Cole model. From total body potassium, intracellular water (ICW) was calculated. In addition, in 14 subjects, total body water (TBW) and extracellular water (ECW) was measured using deuterium oxide dilution and bromide dilution, respectively. Prediction formulas from the literature from impedance at different frequencies were used to predict TBW and ECW, and ICW was calculated as the difference of predicted TBW and ECW. Predicted and measured values of TBW, ECW and ICW did not always show the same values as measured. Generally the ICW from potassium was well correlated with calculated ICW from impedance values, correlation coefficients varying from 0.68 to 0.79, depending on the used prediction formula. From the impedance index H2/Ricf, body weight and age, ICW from potassium was predicted with stepwise multiple regression. The prediction formula was ICW = 0.37065.H2/Ricf - 0.132.age + 0.105.weight + 12.2. The prediction error was 1.9 kg and the explained variance 0.69. The residuals of this prediction formula were dependent on the level of ICW as measured by potassium. The same phenomenon was observed when ICW was calculated as the difference of predicted TBW and ECW. The results show that multifrequency impedance is able to predict ICW, however, the prediction is influenced by body water distribution. PMID- 7486843 TI - Response of the calf pancreas to differently processed soya bean and pea diets. AB - The purpose of this study was to determine the effect of replacing skim-milk powder by differently treated soya bean or pea products on growth, pancreas size and pancreatic enzyme activities in calves. Three separate experiments have been performed. In experiments 1 and 2, 28 and 21 male Holstein calves were divided into 4 or 3 groups, respectively, and fed either dairy products or milk substitutes in which protein was mainly provided by soya bean products differing in their protein concentration due to the technological processing applied. In experiment 3, 45 male Holstein calves were divided into 3 groups and were fed either dairy products, or raw or flaked pea flour as a protein source. After an experimental period of 99 +/- 4 days in experiments 1 and 2, and of 88 days in experiment 3, animal growth rate was significantly lower with raw pea flour (16%) and with the soya bean diet, which was highly concentrated in carbohydrates and allergenic proteins (13-27%). Pancreas weight decreased significantly (16-18%) with pea diets and tended to be lower (NS) with the water extracted, concentrated and heated flour (soya bean). Amylase-specific activity increased significantly (43%) with pea diets but showed opposite tendencies with the most refined soya bean products. Proteolytic enzyme activities were slightly influenced by dietary protein source, but this was not as obvious as in the literature reviewed. Specific messenger RNAs corresponding to amylase, trypsin and chymotrypsin seemed to increase (NS) with the soya bean diets, particularly with the less elaborated one. However, further investigations are required before any conclusions may be drawn concerning regulation levels of pancreatic adaptation to dietary protein. According to this study and the literature, results concerning pancreatic response to diets were different suggesting that the origin of soya bean, pea seeds and technological treatments applied to them were of great importance. Also, the level of incorporation into milk substitute and the presence of more or less antinutritional factors could influence pancreatic enzyme variations by complex mechanisms. PMID- 7486845 TI - Differential effects of geometrical isomers of octadecadienoic acids on ketogenesis and lipid secretion in the livers from rats fed a cholesterol enriched diet. AB - The effect of cis,cis (cc)- and trans,trans (tt)-9,12-octadecadienoic (18:2) acids on ketogenesis and lipid secretion was compared in isolated perfused livers from cholesterol-fed rats. The hepatic uptake of 18:2 acids was comparable in both isomers. The livers perfused with cc-18:2 acid in comparison with those perfused without fatty acid substrate produced approximately 4-fold more ketone bodies accompanying the rise of the beta-hydroxybutyrate:acetoacetate ratio, while the tt-acid isomer further increased these parameters. The hepatic secretion rates of triglyceride and phospholipid as well as cholesterol were all elevated on perfusing the cc-18:2 acid as compared to without fatty acid. In contrast, the rates observed with the tt-18:2 acid isomer except for phospholipid were intermediate, indicating a reciprocal response in ketogenesis and lipid secretion by the trans isomer. The rate of incorporation of trans-fatty acid into perfusate triglyceride and cholesterol ester were lower than cis-acid, but vice versa into perfusate phospholipids. On the other hand, the effects of trans-fatty acid on the concentration and composition of hepatic lipids were less clear. These results emphasize the differential effect of geometrical isomers of the 18:2 acids on oxidation and esterification even in the livers containing a high level of cholesterol. PMID- 7486846 TI - Effects of adaptation to dietary fat on fatty acid composition of serum and serum lipids in miniature swine. AB - The influence of dietary fat on the serum fatty acid composition and distribution in different serum lipids was studied in 24 miniature swine (Sus scrofa) divided into 4 groups. Three groups were fed a low-fat chow with different kind of fats added (sunflower oil, olive oil and bovine lard) and the other group was fed a control diet with no fat added. The animals were fed the four diets for an experimental period of 12 weeks. Afterwards blood samples were taken to obtain serum, and the serum lipids were separated into the following fractions: phospholipids, triglycerides, and esterified cholesterol. The fatty acid composition of the serum and fractions was analyzed by gas chromatography. Under our experimental conditions, the fatty acid composition of the diets was reflected in serum, but not in all cases. In the triglyceride fraction the fatty acid composition was more evident. Our results show the interaction of the fatty acids in the different series and even within a single series, along with the endogenous synthesis processes. PMID- 7486847 TI - 5th International Conference of Anticancer Research. Corfu, Greece, October 17 22, 1995. Abstracts. PMID- 7486848 TI - [Urology in the 20th century]. PMID- 7486849 TI - [Extracorporeal shockwave lithotripsy in children. Study of the effectiveness and renal consequences in a series of eighteen children]. AB - 17 children from 18 months to 16 years old were treated by ESWL for calculi of the upper urinary tract using a Sonolith 3000 lithotriptor. 31 calculi (5 to 60 mm) have been treated on 16 renal units and 3 pelvic ureters. The renal parenchyma of all treated kidneys was considered normal on preoperative ultrasound and IVU; 8 patients had a preoperative DMSA. The treatment required 1 to 4 sessions delivering 400 to 3000 shocks per session and 400 to 6000 shocks per treatment at 14 kV. Within three months post treatment, 14 patients (88.23%) were stone free and 3 patients had residual fragments (2 needed further surgical extraction). With a follow up of three months to five years, all patients have a normal blood pressure, serum creatinine and sterile urine. In no instance, IVU or ultrasound revealed a lesion imputable to ESWL. Seven patients had a DMSA six months or more after the last session; 4 of these were normal but the three other patients had a loss of function and an heterogeneous fixation on the treated side. These preliminary results confirm that ESWL is efficient for upper urinary tract calculi destruction in children. However ESWL seems to be susceptible to induce parenchymal damage, detected by DMSA, whose incidence and long term effects particularly on blood pressure need further evaluation. PMID- 7486851 TI - [Multiple bilateral renal hydatidosis. A case report]. AB - The authors report one case of multiple bilateral hydatid cyst of kidney associated to brain and spleen localizations, after intra cardiac rupture. They point out the rarity of the bilateral kidney localization of hydatidosis, explain the dissemination mode, and stress the role of computed tomography for the pre operative diagnosis. The treatment is above all surgical and adapted to each case. The post-operative results are satisfying. Complementary medical treatment could be useful. PMID- 7486852 TI - [Retroperitoneal liposarcoma. Two cases]. AB - Concerning two cases of liposarcomas developing in the retroperitoneal space, the characteristics of these rare tumors are reviewed. They are the most frequent mesenchymal tumors. Often, they are uncovered by palpable abdominal mass or when they compress adjoining organs. Computed tomography has proved valuable in the diagnosis of these lesions. Surgical removal is the treatment of choice. Recurrences are frequent and prognosis is poor. PMID- 7486850 TI - [Hydatic cyst of the kidney. Eighteen cases]. AB - 18 patients with 18 hydatid cyst of kidney, underwent surgical intervention from 1982 to 1994. There were 6 men in the series and 12 women who ranged in age from 7 to 65 years (mean 38 years). Flank pain was the chief complaint (55%). Abdominal mass was detected on physical examination in 38% of cases. Ultrasonographic examination, performed in all cases, made a correct diagnosis in 13 patients (72%). Excretory urogram (IVP), performed in 9 patients, showed signs of benign tumor process. All patients were operated on: the procedures performed included 12 resection of "protruding dome", 1 total nephrectomy, 3 partial pericystectomy, 2 total pericystectomy, 1 total nephrectomy. In postoperative course, there was one death (5.5%) and morbidity was 11%. We conclude that hydatid cyst of kidney still rare but possible in endemic areas; its treatment, usually surgical, still difficult because it should avoid recurrence of hydatic disease without sacrifice of the kidney. PMID- 7486854 TI - [Rare complications of the double-curved J ureteral stent]. AB - The authors reported two cases of failure of the double-curved ureteral stent. The first is a spontaneous breakage and the second is migration downward and stone formation. The two polyurethane stend had been supported respectively 24 months and 15 months. The first case had been treated with endourology and the second with surgery. PMID- 7486853 TI - [Obstructive anuria. Thirty cases]. AB - The authors report 30 cases of obstructive anuria during the last fifteen years. The anuria was secondary to lithiasis in 60 per cent, in 26.6 per cent to pelvic cancer and in 13.4 per cent to retroperitoneal fibrosis. The diagnosis was facilitated by ultrasonography. Emergency treatment of obstructive anuria is based on urinary diversion by ureteral stent or by percutaneous nephrostomy under ultrasound control. Later the treatment depend of etiology. PMID- 7486855 TI - [Ectopic insertion of a single ureter in the vagina. Value of radiography]. AB - The authors present one case of ectopic ureteric orifice in a vagina in young patient. Clinical investigation suspected a diagnostic. Ultrasonography and computed tomography didn't find renal tissue, but vaginography was most conclusive. The nephroureterectomy brought a cure of disease. PMID- 7486857 TI - [Ureteral stenoses and fistulas after kidney transplantation]. AB - We report a retrospective study of 100 renal transplants, the majority using kidneys from related live donors. We saw 6 ureteric stenoses and four fistulas. Three relatively small stenoses and two limited fistulas were treated percutaneously, all of them successfully. The transplant had to be removed in two patients with a ureteric fistula. Four stenoses and one fistula were treated successfully by open surgery. Actuarial survival of grafts with a urological complication was significantly shorter than that of transplants free of such lesions. PMID- 7486858 TI - [Renal transplantation from related live donors. Results and survival]. AB - We report a retrospective study of 91 renal transplants from related live donors carried out at the Charles Nicolle Hospital, Tunis, between 1986 and 1993. There were 27 HLA-identical and 64 semi-identical. Classical immunosuppression based upon corticosteroids and azathioprine was used in the majority of cases. Surgical complications occurred in 15.4% of cases. 56% of patients had at least one acute rejection attack. There were 4 deaths, 3 related to infection and one to septic arterial rupture. Return to hemodialysis was related to chronic rejection in 5 cases and to recurrence of the initial nephropathy in one case. 5-year actuarial survival was 94% for recipients and 81% for grafts. In this series, acute rejection had no negative influence on graft survival and the difference between graft survival between HLA-identical and semi-identical patients was not statistically significant. PMID- 7486856 TI - [Absence of litholytic effects of zinc sulfate and copper sulfate in experimental lithiasis in rats]. AB - Seventy two male Wistar-strain rats were fed lithogenic diet with ethylene-glycol within three weeks. At the end of this treatment, six rats were killed in order to determine the oxalate and calcium concentrations in renal tissue. Remained rats was randomly divided in four series, each series consisting of three groups. In first series (T), the animals were treated with distilled water; in the second (D1Zn), the animals were treated intramuscularly with the zinc at the rate of 24 micrograms per 100 grams of body weight and per day; in the third (D2Zn), 240 micrograms of zinc were administrated to animals and in the last series (D2Cu), the animals were treated at the same dose as the previous series, but with the copper. The groups which making up each series were killed successively at the 5th, 10th and 15th day after ending treatment with ethyleneglycol in order to determine urinary pH, percentage of water in renal tissue, uremia and concentrations of oxalate and calcium in renal tissue. Then, the comparisons of means were carried out, at each time, between different treated groups and reference group which was treated with distilled water. Litholytic effect was found in all series, including that which had been treated with distilled water. However, compared to reference animals, no acceleration of litholytic process was induced by zinc or copper. The high doses used in these experimentations proved therefore that calcium oxalate calculi were insoluble by zinc and copper. PMID- 7486859 TI - Neuromyotonia: a new autoimmune disease. PMID- 7486860 TI - Tumor necrosis factor: immunogenetics and disease. PMID- 7486861 TI - Inclusion body myositis and myopathies. PMID- 7486863 TI - Genetic control of multiple sclerosis: increased production of lymphotoxin and tumor necrosis factor-alpha by HLA-DR2+ T cells. AB - Lymphotoxin (LT) and tumor necrosis factor-alpha (TNF-alpha) play an important role in the pathogenesis of multiple sclerosis (MS). MS is associated with the HLA-DR2, Dw2, DQ6 HLA class II haplotype. Because both LT and TNF-alpha are encoded in the HLA region, the HLA association of MS may be related to the production of these cytokines. To test this hypothesis, we investigated the production of LT, TNF-alpha, and interferon-gamma (IFN-gamma) by CD4+ T-cell lines (TCLs) specific for myelin basic protein (MBP) or tetanus toxoid (TT) isolated from MS patients and normal controls. After stimulation with specific antigen but not mitogen, TCLs from HLA-DR2+ donors produced significantly more LT and TNF-alpha than TCLs from DR2- donors. In contrast, HLA-DR2+ and DR2- TCLs did not differ in the production of IFN-gamma, a cytokine also produced by T cells but not encoded in the HLA region. Increased secretion of LT and TNF-alpha was unrelated to the specificity (MBP vs TT), MHC restriction (HLA-DR2 vs other DR molecules), or source (MS vs normal) of the TCLs. There was no significant association of the cytokine production with individual LT or TNF-alpha alleles, indicating that the increased production of these cytokines may be linked to other polymorphic genes in this region. The results suggest that the association of MS with HLA-DR2 implies a genetically determined propensity of T cells to produce increased amounts of LT and TNF-alpha. PMID- 7486862 TI - Acquired neuromyotonia: evidence for autoantibodies directed against K+ channels of peripheral nerves. AB - Acquired neuromyotonia is characterized by hyperexcitability of motor nerves leading to muscle twitching, cramps, and weakness. The symptoms may improve following plasma exchange, and injection of immunoglobulin G (IgG) from 1 neuromyotonia patient into mice increased the resistance of neuromuscular transmission to d-tubocurarine. Here we examine nerves and muscle in vitro from mice injected with plasma or purified IgG from 6 neuromyotonia patients or pooled control subjects, and cultured dorsal root ganglion cells after treatment with IgG. Three of the patients had antibodies against human voltage-gated potassium channels labeled with 125I-alpha-dendrotoxin. The quantal release of acetylcholine (quantal content) at end-plates in diaphragms from mice treated with neuromyotonia IgG preparations was increased by 21% relative to control values (p = 0.0053). With one IgG preparation, the duration of the superficial peroneal nerve compound action currents was increased by 93%. The dorsal root ganglion cells treated with this IgG showed a marked increase in repetitive firing of action potentials. All effects were similar to those obtained with aminopyridines. We conclude that at least some patients with acquired neuromyotonia have antibodies directed against aminopyridine- or alpha dendrotoxin-sensitive K+ channels in motor and sensory neurons, and they are likely to be implicated in the disease process. PMID- 7486864 TI - Neurodevelopmental effects of X monosomy: a volumetric imaging study. AB - Thirty girls with Turner syndrome (TuS) were compared with 30 individually age matched controls on volumetric brain measures derived from magnetic resonance imaging and on measures of psychological functioning. As expected, girls with TuS performed more poorly on visual-spatial and intellectual measures relative to controls, and were rated by their parents as having more significant problems in attention and social behaviors. Although no group differences in overall cerebral or subcortical volumes were observed, the regional distribution of gray and white matter differed across groups in both right and left parietal regions. Differences in total tissue volume ratios were seen for both right and left parietal areas, but differences in individual gray and white matter ratios were seen exclusively in the right parietal regions. In general, girls with TuS had a smaller proportion of tissue (gray and white) within the right and left parietal regions, and a larger proportion of tissue within the right inferior parietal occipital region relative to girls in the control group. These data suggest a potentially important role for X chromosome genes and/or sex steroids in the development and specialization of brain structure and function. PMID- 7486865 TI - Cortical control of double-step saccades: implications for spatial orientation. AB - To accurately localize a visual target in space despite eye movement-induced shifts of its retinal image, the brain must take into account both its retinal location and information about current eye position or at least the preceding eye displacement. We examined this ability with respect to saccadic eye movements by applying "double-step" stimuli, where the locations of two sequentially flashed target lights have to be fixated by two successive saccades performed after their disappearance. As the 2nd saccade will not start at the spatial location from which the 2nd target was seen, a dissonance arises between its retinal coordinates and the motor coordinates of the required 2nd saccade. Nevertheless, these saccades were performed quite accurately by 32 healthy human adults. To investigate the contribution of the cerebral cortex, we recorded horizontal double-step saccades in 35 patients with focal unilateral hemispheric lesions. Whereas frontal lesions impaired temporal properties, posterior parietal lesions caused spatial dysmetria or failure of even ipsiversive 2nd saccades following contraversive 1st saccades. This reflects an inability to compensate for retinospatial dissonance by using nonretinal information (corollary discharge) about eye displacement associated with a previous saccade into the contralesional hemifield. In conclusion, the parietal cortex is crucial for spatial constancy across saccades. PMID- 7486866 TI - Resolution-dependent estimates of lesion volumes in magnetic resonance imaging studies of the brain in multiple sclerosis. AB - In this study, we investigated the relationship between multiple sclerosis lesion volumes measured from magnetic resonance imaging scans and image-slice thickness. The lesion volume was computed using a semiautomated thresholding technique from axial scans of the brain of varying slice thickness. Ten patients were studied, and in all cases the computed lesion volume increased with decreasing slice thickness (p = 0.01). Linear extrapolation from our data allowed the lesion volume at very small slice thickness to be estimated; this was found to be on average 20% greater than that obtained using a slice thickness of 5 mm. Furthermore, there were considerable differences in the percentage of change in lesion volume from patient to patient, and it would appear that there is a larger variation with slice thickness for patients with smaller lesions and higher lesion loads. PMID- 7486868 TI - Significance of spikes recorded on electrocorticography in nonlesional medial temporal lobe epilepsy. AB - Whether spikes recorded by intraoperative electrocorticography imply active epileptogenicity has not been adequately addressed. We performed preresection and postresection electrocorticography on 47 patients with nonlesional medial temporal lobe epilepsy who were undergoing surgery for the treatment of medically refractory epilepsy. A standard anteromedial temporal lobectomy was performed on all patients, with no additional resection, regardless of electrocorticographic findings. Patients were divided into two groups: Group I (no seizures or rare seizures) and Group II (recurrent seizures). Recorded spikes were analyzed for distribution and spike discharge rate. On preresection electrocorticography, 83% of Group I and 82% of Group II had spikes in the anterior temporal lobe. The spike discharge rate was equally distributed between high frequency and low frequency for both groups (not significant). Although spikes localized to the posterior temporal neocortex were seen more in Group II (64%) than Group I (39%), this was not a significant difference (p > 0.1). Most of these patients had a low frequency spike discharge rate. On postresection electrocorticography, 80% of Group I and 75% of Group II had residual spikes. The majority of these had a low frequency spike discharge rate and were localized to the margin of resection. We found no correlation between residual spikes on preresection and postresection electrocorticography and outcome. These findings do not support the role of intraoperative electrocorticography in guiding mesial temporal lobe resection. PMID- 7486867 TI - Immunocytochemical quantitation of human immunodeficiency virus in the brain: correlations with dementia. AB - The pathogenesis of human immunodeficiency virus (HIV)-associated dementia is unclear, and the underlying pathological substrate has been a matter of debate. In a prospectively clinically characterized population of acquired immunodeficiency syndrome (AIDS) patients we investigated the relationship between the clinical syndrome of HIV-associated dementia and the presence and relative quantity of immunocytochemical markers for HIV-1 (gp41 antibody), and for macrophages and microglia (HAM-56 antibody). Sections from the basal ganglia and frontal lobes from the brains of 51 patients were studied, and the data were stratified for severity of dementia (16 nondemented, 12 mildly demented, 23 severely demented), rate of dementia progression, duration of AIDS, use of antiretrovirals, and several other demographic features. We found a highly significant correlation between the degree of macrophage staining and the severity of dementia but only a borderline correlation between the presence and amount of gp41-positive cells and dementia. Several nondemented patients showed abundant gp41 immunoreactivity, and some severely demented showed little to no gp41 immunoreactivity. Other correlations with the immunostaining data, including antiretroviral use, were not significant. We conclude that the presence of macrophages and microglia is a better correlate with HIV-associated dementia than is the presence and amount of HIV-infected cells in the brain. These data support the concept that the pathogenesis of HIV-associated dementia is likely due to indirect effects of HIV infection of the brain, possibly through the actions of macrophages and microglia. PMID- 7486869 TI - The effect of deprenyl and levodopa on the progression of Parkinson's disease. AB - We have performed a 14-month, prospective, randomized, double-blind, placebo controlled study to evaluate the effect of deprenyl and levodopa/carbidopa (Sinemet) on the progression of signs and symptoms in patients with mild Parkinson's disease (PD). One hundred one untreated PD patients were randomly assigned to one of the following four treatment groups: Group I, deprenyl + Sinemet; Group II, placebo-deprenyl + Sinemet; Group III, deprenyl + bromocriptine; and Group IV, placebo-deprenyl + bromocriptine. The final visit was performed at 14 months, i.e., 2 months after withdrawal of deprenyl or its placebo and 7 days after withdrawal of Sinemet or bromocriptine. Deterioration in Unified Parkinson's Disease Rating Score (UPDRS) between untreated baseline and final visits was used as an index of disease progression. Placebo-treated patients deteriorated by 5.8 +/- 1.4 points, while deprenyl-treated patients deteriorated by 0.4 +/- 1.3 points (p < 0.001). This effect was sufficiently powerful that a significant deprenyl effect could be detected in the subgroup of 41 patients randomized to Sinemet (p < 0.01) as well as in the 23 patients who completed a 14-day washout of Sinemet or bromocriptine (p < 0.05). No difference in the extent of deterioration was detected in patients randomized to Sinemet versus bromocriptine. This study demonstrates that deprenyl attenuates deterioration in UPDRS score in patients with early PD. These findings are not readily explained by the drug's symptomatic effects and are consistent with the hypothesis that deprenyl has a neuroprotective effect. PMID- 7486870 TI - Depth electrode studies and intracellular dentate granule cell recordings in temporal lobe epilepsy. AB - Hippocampal depth electrodes are often used to localize seizure onset in patients who may have temporal lobe epilepsy (TLE). A number of features of the spontaneous seizures and of their ictal onset patterns can be analyzed from these recordings. We compared a number of the typical electroencephalographic (EEG) changes at seizure onset with several cellular parameters recorded in dentate granule cells from the same 14 patients diagnosed with medial temporal sclerosis (MTS) to examine the pathophysiological correlates of this spontaneous EEG activity in this form of TLE. The intracellularly recorded parameters include the propensity to fire evoked epileptiform bursts, the absence of evoked inhibitory potentials, the presence of polysynaptic excitatory postsynaptic potentials, and the presence of spontaneous excitatory activity. We noted several correlations between the EEG data and the intracellular recordings. The absence of synaptically evoked bursts was correlated with the presence of low-voltage fast activity at seizure onset. In addition, the loss of inhibitory postsynaptic potentials was correlated with the presence of periodic spiking pre-ictally. Several other correlations were also noted. These data indicate that EEG findings may be predictive of anatomical and cellular pathological changes and provide clues to the physiological mechanisms involved in this form of epilepsy. PMID- 7486871 TI - Monocyte/macrophage differentiation in early multiple sclerosis lesions. AB - Monocyte/macrophage differentiation was studied in biopsy samples of multiple sclerosis (MS) lesions obtained in the early course of the disease. Macrophages were identified by immunocytochemistry using a panel of antibodies recognizing different macrophage-activation antigens. The number of cells stained with each antibody was related to the demyelinating activity of the lesions as detected by the presence of myelin degradation products. The pan-macrophage marker Ki-M1P revealed the highest numbers of macrophages in early and late active lesions. Lower numbers were encountered in inactive, demyelinated, or remyelinated lesions. The acute stage inflammatory macrophage markers MRP14 and 27E10 were expressed in either only early active (MRP14) or early and late active (27E10) lesions, thus allowing the identification of actively demyelinating lesions. The chronic stage inflammatory macrophage marker 25F9, in contrast, showed increasing expression with decreasing lesional activity. These findings indicate a differentiated pattern of macrophage activation in MS lesions and allow the staging of demyelinating lesions in routinely fixed and paraffin-embedded tissue. PMID- 7486872 TI - Apolipoprotein E genotype in patients with Alzheimer's disease: implications for the risk of dementia among relatives. AB - Numerous studies have shown that the risk of Alzheimer's disease (AD) is associated with the dose of the epsilon 4 allele of apolipoprotein E (ApoE). However, more than one third of AD patients lack epsilon 4 and many persons having epsilon 4 survive cognitively intact to old age. We evaluated the lifetime risk of disease in 3,999 first-degree relatives of 549 probands who met the criteria for probable or definite AD and whose ApoE genotypes were known. ApoE genotypes for relatives were not determined. After age 65 the risk among relatives was proportional, as much as 7 to 10% at age 85, to the number of epsilon 4 alleles present in the proband. Risks to relatives of ApoE 2/2 and 2/3 probands were nearly identical at all ages to risks for relatives of ApoE 3/3 probands. The expected proportion of relatives having at least one epsilon 4 allele was calculated for each genotype group based on the distribution of parents, sibs, and offspring in the sample. Among relatives in the ApoE 3/3 group, the lifetime risk for AD by age 90 was three times greater than the expected proportion of epsilon 4 carriers, suggesting that factors other than ApoE contribute to AD susceptibility. Furthermore, the 44% risk of AD by age 93 among relatives of ApoE 4/4 probands indicates that as many as 50% of people having at least one epsilon 4 allele do not develop AD. We also found that among male relatives, risk of AD in the ApoE 3/4 group was similar to that for the ApoE 3/3 group but significantly less than the risk for the ApoE 4/4 group. In contrast, among female relatives the risk for the ApoE 3/4 group was nearly twice that for the ApoE 3/3 group and identical to the risk for the ApoE 4/4 group. These findings are consistent with a sex-modification effect of the E4 isoform on disease susceptibility. PMID- 7486873 TI - Anti-ganglioside GM1 antibodies in Guillain-Barre syndrome and their relationship to Campylobacter jejuni infection. AB - To clarify the association between Campylobacter jejuni (Cj) infection and antibodies to ganglioside GM1 (anti-GM1) in Guillain-Barre syndrome (GBS), we have carried out a prospective case-control study of 96 patients with GBS. Cj infection occurred in 25 (26%) patients. IgG and/or IgM anti-GM1 were identified in 24 (25%) patients and in 1 of 71 (1.4%) household controls (p < 0.001). Thirteen of the 25 (52%) Cj-positive patients had anti-GM1 compared with 11 of the 71 (15%) Cj-negative patients (p < 0.001). Neither the peak overall disability nor the 1-year disability differed between the anti-GM1-positive and anti-GM1-negative patients. However, patients with the combination of Cj infection and anti-GM1 positivity recovered more slowly than Cj/anti-GM1-negative patients (p = 0.05), were more likely to have axonal degeneration, and were significantly more disabled at the end of 1 year (p = 0.02). The presence of Cj infection is more important than anti-GM1 positivity in determining the extent of axonal involvement and, hence, prognosis. Since the presence of anti-GM1 is not a significant poor-prognostic factor, a search should be made for other properties of Cj infection that would account for its relationship to axonal degeneration. PMID- 7486874 TI - Familial hemiplegic migraine and autosomal dominant arteriopathy with leukoencephalopathy (CADASIL). AB - Cerebral autosomal dominant arteriopathy with subcortical infarcts and leukoencephalopathy (CADASIL) is a recently described familial cerebrovascular disorder shown to map to chromosome 19q12. Familial hemiplegic migraine has also been shown in some families to map close to the CADASIL locus. The fully developed CADASIL phenotype consists of recurrent strokes developing in the fourth decade, progressing to a pseudobulbar palsy, spastic quadriparesis, and subcortical dementia. In an Irish family 15 members were fully investigated by magnetic resonance scanning; 10 had typical magnetic resonance features of CADASIL. Five members of this family had familial hemiplegic migraine and 4 of these had magnetic resonance evidence of CADASIL. Two other members had migraine with and without aura as a presenting clinical symptom of CADASIL. This disorder has been shown by linkage analysis to map to the CADASIL locus at chromosome 19. The phenotype at presentation of CADASIL in this family was variable and age related and included familial hemiplegic migraine, migraine with and without aura, transient ischemic attacks, strokes, and spinal cord infarction. This family study increases our understanding of the spectrum of clinical manifestations of this underrecognized familial cerebrovascular disorder. PMID- 7486875 TI - Neurological sequelae of cyanide intoxication--the patterns of clinical, magnetic resonance imaging, and positron emission tomography findings. AB - We report 2 patients with neurological sequelae of oral cyanide intoxication who were evaluated clinically and neuropsychologically, with high-resolution magnetic resonance imaging and one of them with positron emission tomography. The clinical syndrome was characterized by extrapyramidal motor and cerebellar symptoms such as bradykinesia, hypomimia, slowed speech, anteropulsion, and marked retropulsion, but little tremor. The sensory and pyramidal motor systems were normal or relatively spared. On neuropsychological testing the intellectual capacity and memory functions were normal, whereas the speed of motor reaction and verbal fluency were reduced. Magnetic resonance imaging showed damage of the globus pallidus, putamen, substantia nigra, subthalamic nucleus, and cerebellum in both patients, whereas the sensory-motor cortex and hippocampus were relatively spared. 18F-6-Fluoro-dopa positron emission tomography revealed a symmetrical reduction of striatal dopa uptake by 42% on average that was similar in the putamen and caudate. 18F-2-Fluoro-2-deoxyglucose positron emission tomography showed a regional reduction of the glucose metabolism in the posterior putamen and temporo-parieto-occipital and cerebellar cortex. PMID- 7486876 TI - Chronic inflammatory demyelinating polyneuropathy complicating liver transplantation. AB - We report on a patient who had chronic inflammatory demyelinating polyradiculoneuropathy following liver transplantation for hepatitis B-induced fulminate hepatic failure. Possible symptomatic recurrence of hepatitis B in the graft prompted a reduction in the cyclosporine dosage, which may have triggered the immune response leading to demyelination. Initial plasma exchange and then intravenous immune globulin after relapse led to marked clinical improvement. PMID- 7486877 TI - Biased assessment of blinding in a randomized placebo-controlled trial of oral methotrexate in chronic progressive multiple sclerosis. PMID- 7486878 TI - German population data on the loci low-density-lipoprotein receptor, glycophorin A, hemoglobin gamma G, D7S8 and group-specific component. AB - This paper reports on the allele and genotype frequencies for five polymerase chain reaction-based loci determined in a sample (n = 295) from Dusseldorf (Germany). The loci are Low-Density-Lipoprotein Receptor, Glycophorin A, Hemoglobin gamma G, D7S8 and Group-Specific Component. Typing was carried out by using the AmpliType PM PCR amplification and typing kit. The loci were tested for possible divergences from Hardy-Weinberg expectations (HWE). The gene frequencies found can be used in forensic analyses and paternity tests. The data from Dusseldorf were compared with six other studies having been published up today. PMID- 7486879 TI - [Distribution of ABO allele frequencies in the Cologne registration district (Nordrhein-Westfalen)]. AB - The AB0 allele frequencies and their regional distribution have been studied in the administrative area of Koln, North Rhine-Westphalia. The B allele shows relatively high frequencies ( > 0.0800) on both sides of the river Rhein, in the eastern parts of the area and--separated from the Koln region by a region of lower B frequencies--in the Aachen region. The possible causes for the different B frequencies are discussed. PMID- 7486880 TI - Immune-genetic study of the new populations of Sierra Morena (Spain) through HLA polymorphisms of class I. AB - We have studied the HLA antigenes of class I in 50 families from Andalusia and in 50 families from some towns of Sierra Morena (La Carolina among others), in which an important settlement took place by individuals from Central Europe over 200 years ago, by comparing the allele frequencies with the German population. The obtained results allowed us to conclude that a real mixture of population has occurred. The frequencies of some HLA alleles of class I in the population of Sierra Morena are intermediate to those observed in the original populations, though some phenomena of drifting have been noticed. The study of gametic associations also shows that the population of Sierra Morena behaves as an intermediate population between the two original populations. This is evident in relation to the A1-B8 haplotype, in which the population of Sierra Morena is similar to the original Andalusian population, regarding the frequency of the haplotype, and the German population, because of the intensity of disequilibrium. The study of the genetic distances shows that the ones existing between the populations of Andalusia, Germany and Sierra Morena are small, although they reveal that these populations are clearly defined. PMID- 7486881 TI - Distribution of HLA antigens in Sadhu Chetty of Tamil Nadu, south India. AB - Blood samples from 101 individuals of Sadhu Chetty community domiciled in and around Madras (capital City of Tamil Nadu State, South India) were examined for HLA-A, B, C, DR and DQ antigen profiles. Phenotype, gene and haplotype frequencies were calculated and compared with the literature. Increased frequencies of HLA-B16 antigen and of the haplotype A2, B16 were characteristic. This study indicates the distinctiveness of the Sadhu Chetty population and highlights the importance of determining HLA frequencies in endogamous groups of India. PMID- 7486882 TI - Regulation of food intake during the menstrual cycle. AB - Several studies indicate that food intake and selection varies during menstrual cycle. The present results are based on nutritional diaries kept by 27 healthy regularly menstruating women, who did not use oral contraceptives. Over a time period of at least one menstrual cycle the women recorded their daily ad libitum food intake by specifying the different nutrients and the respective amounts. The daily total energy, carbohydrate, protein and fat intake was analyzed using the national foodstuff code provided by the National Health Institute with regard to different phases of the menstrual cycle. The latter were controlled by daily measurements of basal temperature. The results indicate that during the luteal phase of menstrual cycle the intake of total energy tends to be higher than during follicular phase. Carbohydrate intake was on the lowest point during periovulatory period reaching a minimum approximately 2 days after ovulation. Such a minimum could also be observed in fat and total energy intake. Furthermore it could be demonstrated that the subjective perception of "sweetness" varied with cycle phases. The results suggest that food intake and selection is influenced by neurochemical, hormonal, physiological and psychological factors. PMID- 7486883 TI - Complex statistical characterization of women's body measurements. AB - 670 young Estonian female students of the Tartu University (aged 18-22 years) were examined anthropometrically (37 body measurements and 10 skinfolds). By means of multidimensional statistical analysis we found that the woman's body as a whole is a linearly well-correlated system. The leading measurements are height and weight, which account for 50% of the variability of all the other measurements, while individual variability makes up 50%. There are no other body measurements besides weight and height that can give a reliable description of the general build of a woman's body. Variations in body height-weight sizes lead to systematic changes in the length, breadth and depth measurements, circumferences and body proportions. Comparative changes of body proportions in the general contingent and in the groups of purely pyknic and leptosomic women are based on the corresponding values of their body height and weight. These findings lead to the conclusion that the anthropological whole body primary model may be a classification of women's body height and weight. PMID- 7486884 TI - [Gladebeck--an early medieval cemetery in Lower Saxony]. AB - The cemetery of Gladebeck is placed in the South of Niedersachsen and dated from the 8th till the early 9th century. The 55 individual findings consist of 17 men, 16 women, 11 undeterminables and 11 children. The mean death rate of the whole population is 29 years. The age-distribution shows a deficiency of small infants. PMID- 7486885 TI - A study of leg-crossing in a German sample. AB - When a person is asked to cross their flexed legs then one thigh crossed over the other. This paper reviews the previous literature on leg crossing, and reports new data. In this study about 62% of the population (n = 292) are right leg crossers, 26% are left leg-crossers, and the remaining 12% report that they have no preference or are indifferent. The incidence of left leg-crossing differs between the sexes, being higher in males, although there seems to be no association with age. Leg-crossing is correlated to some extent with footedness, handedness and eyedness. PMID- 7486886 TI - Incidence, density and direction of hair on the dorsum of phalanges of the hands in Malays. AB - In this study 200 Malay subjects (100 males and 100 females) were randomly selected from patients attending outpatient clinics of Hospital USM, Kelantan, Malaysia, to find out the incidence, density and direction of hair on the dorsum of phalanges of the hand. These features have not been studied so far in females nor has such a study been conducted in Malays. The probability of density of hair distribution among the digits of both hands showed significant correlation on proximal phalanges (p < 0.05) in both sexes. Significant correlation was not observed, however, in the middle phalangeal hair (MPH) of the hands. The direction of proximal phalangeal hair, from little finger to the thumb, showed significant changes from ulnar to radial in both sexes (p < 0.05). Identification of isolated digits, which is of medicolegal importance, would be more accurate if the direction and the density of hair on the digits are both considered together. MPH was present in 48% of males and in 33% of females studied. Comparisons with presence of MPH in other populations show that Malays are ethnically similar to other Asiatic populations. PMID- 7486887 TI - [Voluntary, controlled limited reproduction in humans--a conflict to modern evolutionary biology?]. AB - In western industrialized countries it is widespread to limit individual reproduction voluntarily and deliberately. This phenomenon of minimizing reproduction conflicts with the maximization of biological fitness, expected on the basis of sociobiology. Some positions and attitudes of demography and sociobiology that appear to be contradictive are critically discussed, and with several examples it is expounded that we are dealing with misunderstanding rather than inconsistencies in many cases. The majority of misunderstandings can be put down to an inadequate differentiation between proximate and ultimate levels of causation. However, proximate interpretations of the problem of minimizing reproduction from a sociobiological perspective are frequently unsatisfactory. Therefore, the demographic model of "competing pleasures of life" (Konkurrenz der Genusse) was connected with evolutionary biology. Combining demographic and biological aspects reveals that human striving for material and non-material values (pleasures of life), originally was highly adaptive and fitness-enhancing as behaviors to acquire resources and secure social status. Not until a society offered a variety of behavioral and consumer alternatives and simultaneously sophisticated methods of contraception these behavioral patterns dissociated from their function of survival and became independent. PMID- 7486888 TI - From Carper's patterns of knowing to ways of being: an ontological philosophical shift in nursing. AB - Carper's 1978 article in the premiere issue of Advances in Nursing Science encouraged nurses to consider four fundamental patterns of knowing. Through illustrations from literature and the performing arts, the authors address Carper's patterns of knowing in the context of an emerging philosophical shift. First, they critique the major strengths and limitations of the article. Next, they explore an emerging philosophical shift in nursing from Carper's epistemological focus to ontological reflections on ways of being. Finally, they discuss the significance of the emerging philosophical shift and the ways of being for the science-art of nursing. PMID- 7486889 TI - Stories and numbers: coexistence without compromise. AB - Research methods are not paradigm specific but should be selected on the basis of whether they fit with the purposes of an investigation. In the postpositivist, interpretive, and critical paradigms, both qualitative and quantitative data or a combination of these may be used without violating paradigm assumptions. Attention to the four basic issues of quality of the data, investigator bias, quality of the research process, and usefulness of the finding is necessary to produce valid research. Although researchers in each paradigm deal with these differently, combining strategies across paradigms may enhance the scientific value of a study and result in new methodologies to address the health needs of all people. PMID- 7486890 TI - "The focus of the discipline of nursing": a critique and extension. AB - Influential writings in nursing tend to align with views held and privileged in the scientific community. By taking this stance nurses resist changes that would advance the discipline and give voice and visibility to marginalized people in society who seek the socially relevant service that nursing purports to offer. The difficulty stems not only from the failure to question the dominance of particular approaches for knowledge generation over others, but also from the failure to recognize that our questions are constructed to support rather than to challenge existing knowledge. Revisioning a pivotal article--"The focus of the discipline of nursing"--enhances its usefulness for nursing curricula. PMID- 7486891 TI - "Methodological notes on empathy": further considerations. AB - This article updates and extends Jeannette Gagan's "Methodological notes on empathy," published in the January 1983 issue of ANS. The evolving conceptualization of empathy as a complex, intrapsychic, and interpersonal process suggests that the inconclusiveness and apparent contradictions of research to date might be substantially clarified by considering data from a multidimensional perspective. Attention to the conceptual bases underlying diverse measurement approaches is key to understanding the data produced. Notwithstanding the limitations of their paradigms and models for nursing, the debates and knowledge accumulated in other disciplines can inform a broad range of inquiry relevant to nursing. PMID- 7486892 TI - Older women and their images of health: a replication study. AB - Older women view themselves as healthy despite the fact that they suffer from chronic illnesses and some functional limitations. The study presented in this article replicates the work of Woods and associates by asking, "What does being healthy mean to you?" to a sample of 10 older women aged 70 to 91 years. The images presented by older women were remarkably consistent with the images presented by the younger cohort. As found in the original Woods et al study, there were nine dimensions that were reflective of the eudaemonistic model of health. PMID- 7486893 TI - Evaluation of the Interaction Model of Client Health Behavior through the first decade of research. AB - The Interaction Model of Client Health Behavior was proposed in 1982 as a prescriptive nursing framework. The model is grounded in multidisciplinary perspectives, and early research suggests that the model may be useful for research and practice. The elements of client singularity and health outcomes have been the primary focus of research during the past decade. Statistical analyses have been exploratory. This article concludes that the model is ready for hypothesis testing with more rigorous statistical procedures, more diversity in samples, and more comprehensive use of the model, including the elements of client-professional interaction. PMID- 7486894 TI - Beige mouse model for Mycobacterium avium complex disease. PMID- 7486895 TI - Comparison of sparfloxacin, temafloxacin, and ciprofloxacin for prophylaxis and treatment of experimental foreign-body infection by methicillin-resistant Staphylococcus aureus. AB - The prophylactic and therapeutic activities of three broad-spectrum fluoroquinolones were evaluated in two different experimental models of foreign body infections caused by methicillin-resistant Staphylococcus aureus (MRSA) susceptible to quinolones. In a guinea pig model of prophylaxis, subcutaneously implanted tissue cages were infected at a > 90% rate by 10(2) CFU of MRSA in control animals. A single dose of 50 mg of ciprofloxacin per kg of body weight administered intraperitoneally 3 h before bacterial challenge was less effective than an equivalent regimen of either sparfloxacin or temafloxacin in decreasing the rate of experimental infection in tissue cages challenged with increasing inocula of MRSA. In a rat model evaluating the therapy of chronic tissue cage infection caused by MRSA, the efficacy of a 7-day high-dose (50-mg/kg twice daily) regimen of sparfloxacin, temafloxacin, or ciprofloxacin was compared to that of vancomycin (50 mg/kg twice daily). Active levels of sparfloxacin, temfloxacin, or ciprofloxacin were continuously present in tissue cage fluid during therapy, exceeding their MBCs for MRSA by 6- to 20-fold. Either temafloxacin, sparfloxacin, or vancomycin was significantly (P < 0.01) more active than ciprofloxacin in decreasing the viable counts of MRSA in tissue cage fluids. The different activities of ciprofloxacin compared with those of the other two quinolones against chronic tissue cage infections caused by MRSA did not involve the selective emergence of quinolone-resistant mutants. Temafloxacin and ciprofloxacin, which showed the most prominent differences in their in vivo activities, however, exhibited similar bactericidal properties and pharmacokinetic parameters in the rat model. In conclusion, both temafloxacin and sparfloxacin were significantly more active than ciprofloxacin for the prophylaxis or treatment of experimental foreign-body infections caused by a quinolone-susceptible strain of MRSA. PMID- 7486896 TI - Inhibitory and bactericidal activities of levofloxacin, ofloxacin, erythromycin, and rifampin used singly and in combination against Legionella pneumophila. AB - The susceptibilities of 56 Legionella pneumophila isolates (43 clinical and 15 environmental isolates) to levofloxacin, ofloxacin, erythromycin, and rifampin were studied with buffered charcoal yeast extract (BCYE) agar (inoculum, 10(4) CFU per spot), and the susceptibilities of five isolates were studied with buffered yeast extract (BYE) broth (inoculum, 10(5) CFU/ml). The MICs inhibiting 90% of strains tested on BCYE agar were 0.125, 0.25, 1.0, and < or = 0.004 micrograms/ml for levofloxacin, ofloxacin, erythromycin, and rifampin, respectively. The MICs by the BYE broth dilution method were 1 to 3, 2, 1 to 2, and 1 tube lower than those by the agar dilution method for levofloxacin, ofloxacin, erythromycin, and rifampin, respectively. The MBCs were 1 to 2 tubes higher than the broth dilution MICs for levofloxacin, 1 to 3 tubes higher than the broth dilution MICs for ofloxacin, 1 to 3 tubes higher than the broth dilution MICs for erythromycin, and the same as the broth dilution MICs for rifampin. In kinetic time-kill curve studies, at drug concentrations of 1.0 and 2.0 times the MIC, the most active drugs were levofloxacin and rifampin. At 72 h, concentrations of levofloxacin and rifampin of 2.0 times the MIC demonstrated a bactericidal effect against L. pneumophila. In contrast, at concentrations of 1.0 and 2.0 times the MICs regrowth was observed with ofloxacin and only a gradual decrease in the numbers of CFU per milliliter was observed with erythromycin. Only a minor inhibitory effect was observed with 0.25 or 0.5 time the MICs of all drugs at 24 to 48 h, with regrowth occurring at 72 h. In contrast to erythromycin or ofloxacin plus rifampin at 0.25 time the MICs, only levofloxacin plus rifampin demonstrated synergy. Thus, levofloxacin demonstrated the best inhibitory and bactericidal effects against L. pneumophila when it was studied alone or in a combination with rifampin. PMID- 7486898 TI - Effects of an acidic beverage (Coca-Cola) on absorption of ketoconazole. AB - Absorption of ketoconazole is impaired in patients with achlorhydria. The purpose of this study was to determine the effectiveness of a palatable acidic beverage (Coca-Cola Classic, pH 2.5) in improving the absorption of ketoconazole in the presence of drug-induced achlorhydria. A prospective, randomized, three-way crossover design with a 1-week wash-out period between each treatment was employed. Nine healthy nonsmoking, nonobese volunteers between 22 and 41 years old were studied. Each subject was randomized to receive three treatments: (A) ketoconazole 200-mg tablet with water (control), (B) omeprazole (60 mg) followed by ketoconazole (200 mg) taken with water, and (C) omeprazole (60 mg) followed by ketoconazole (200 mg) taken with 240 ml of Coca-Cola Classic. The pH values of gastric aspirates were checked after omeprazole was administered to confirm attainment of a pH of > 6. Multiple serum samples were obtained for measurements of ketoconazole concentrations by high-pressure liquid chromatography. The mean area under the ketoconazole concentration-time curve from zero to infinity for the control treatment (17.9 +/- 13.1 mg.h/liter) was significantly greater than that for treatment B (3.5 +/- 5.1 mg.h/liter; 16.6% +/- 15.0% of control). The mean peak concentration was highest for the control treatment (4.1 +/- 1.9 micrograms/ml), for which the mean peak concentration showed a significant increase over that for treatment B. The absorption of ketoconazole was reduced in the presence of omeprazole-induced achlorhydria. However, drug absorption was significantly increased, to approximately 65% of the mean for the control treatment, when the drug was taken with an acidic beverage, such as Coca-Cola. PMID- 7486899 TI - Role of extracellular calcium in in vitro uptake and intraphagocytic location of macrolides. AB - We compared the uptakes and intracellular locations of four 14-membered-ring macrolides (roxithromycin, dirithromycin, erythromycin, and erythromycylamine) in human polymorphonuclear neutrophils (PMNs) in vitro. Intracellular location was assessed by cell fractionation and uptake kinetics in cytoplasts (granule-poor PMNs). Trapping of dirithromycin within PMN granules (up to 80% at 30 min) was significantly more marked than the intracellular trapping of the other drugs (erythromycylamine, 45% +/- 5.1%; erythromycin, 42% +/- 3.7%; roxithromycin, 35% +/- 3.0%). A new finding was that, in the absence of extracellular calcium, the uptakes of all of the macrolides by PMNs and cytoplasts were significantly impaired, by about 50% (PMN) and 90% (cytoplasts). Furthermore, inorganic Ca2+ channel blockers inhibited macrolide uptake in a concentration-dependent manner, with 50% inhibitory concentrations of 1.6 to 2.0 mM and 29 to 35 microM, respectively, for Ni2+ and La3+. The intracellular distributions of the drugs were unchanged in the presence of Ni2+ and La3+ and in Ca(2+)-free medium supplemented with ethylene glycol-bis(beta-aminoethyl ether)-N,N,N',N' tetraacetic acid. The organic Ca2+ channel blocker nifedipine had no effect on macrolide uptake, whereas verapamil inhibited it in a time- and concentration dependent manner. These data show the importance of extracellular Ca2+ in macrolide uptake by phagocytes and suggest a link with Ca2+ channels or a Ca2+ channel-operated mechanism. PMID- 7486897 TI - Detection of gyrA and gyrB mutations in quinolone-resistant clinical isolates of Escherichia coli by single-strand conformational polymorphism analysis and determination of levels of resistance conferred by two different single gyrA mutations. AB - Twelve quinolone-resistant clinical isolates of Escherichia coli (nalidixic acid MICs, 64 to 512 micrograms/ml; norfloxacin MICs, 0.25 to 8 micrograms/ml) were transformed with plasmid pJSW101 carrying the gyrA+ gene and with plasmid pJB11 carrying the gyrB+ gene to examine the proportion of gyrA and gyrB mutations. Transformation with pJSW101 resulted in complementation (nalidixic acid MICs, 4 to 32 micrograms/ml; norfloxacin MICs, 0.06 to 0.25 micrograms/ml). In contrast, no change in MICs were observed after transformation with pJB11. A 418-bp fragment of gyrA from the 12 strains was amplified by PCR. Direct DNA sequencing of that fragment identified the causes of quinolone resistance in eight strains as a single point mutation leading to a substitution of the serine at position 83 (Ser-83) to Leu and in four strains as a single point mutation leading to a substitution of Asp-87 to Gly. Exchange of the fragment from one of these strains with that of gyrA+ and transformation of resistance with the hybrid gyrA plasmid indicated the contribution of Gly-87 to resistance and the stabilities of mutants containing GyrA (Gly-87). Thus, gyrA gene mutations are probably encountered more often than gyrB gene mutations in clinical isolates of E. coli. In addition, the substitution of Asp-87 to Gly can be encountered in such strains. On the basis of the level of resistance found in the fragment exchange experiment, the quinolone resistance attributable to Gly-87 appears to be comparable to that attributable to Leu-83. The levels of resistance found in the clinical isolates shown to have a Gly-87 mutation (nalidixic acid MICs, 64 to 512 micrograms/ml; norfloxacin MICs, 0.5 to 4 micrograms/ml) suggest that the Gly-87 mutation causes resistance at the level of the nalidixic acid MIC (64 micrograms/ml) or the norfloxacin MIC (0.5 micrograms/ml or less) and that the additional increments in resistance seen in the other strains with higher levels of resistance may be attributable to additional mutations. The single-strand conformational polymorphism analysis with PCR products readily detected te Leu-83 and Gly-87 mutations. PMID- 7486901 TI - Effect of clarithromycin on sputum production and its rheological properties in chronic respiratory tract infections. AB - Macrolide antibiotics possess a variety of actions other than antimicrobial activities. To determine the effects of long-term administration of clarithromycin (CAM) on the amount and physical properties of sputum in patients with clinical conditions associated with excessive airway secretions, we conducted the present study in a parallel, double-blind, placebo-controlled fashion. Patients were divided into two groups: the first group (n = 16) received CAM (100 mg, twice a day) for 8 weeks, and the second group (n = 15) received placebo. In evaluating airway secretion, the daily amount of expectorated sputum, solid composition, viscoelastic properties (including elastic modulus and dynamic viscosity), and sputum microbiology were assessed. CAM decreased sputum production from 51 +/- 6 to 24 +/- 3 g/day after treatment, whereas placebo had no effect. The bacterial density and sputum flora were unaltered. In the group receiving CAM, the percent solid composition and elastic modulus increased from 2.44% +/- 0.29% to 3.01% +/- 0.20% and 66 +/- 7 to 87 +/- 8 dyne/cm2 (P < 0.05), respectively, but the dynamic viscosity remained unchanged. These results suggest that long-term treatment with CAM reduces the amount of sputum production, probably by inhibiting airway secretions, and increases sputum elasticity. PMID- 7486900 TI - Determination of robust ocular pharmacokinetic parameters in serum and vitreous humor of albino rabbits following systemic administration of ciprofloxacin from sparse data sets by using IT2S, a population pharmacokinetic modeling program. AB - Robust determination of the concentration-time profile of anti-infective agents in certain specialized compartments is often limited by the inability to obtain more than a single sample from such a site in any one subject. Vitreous humor and cerebrospinal fluid are obvious examples for which the determination of concentrations of anti-infective agents is limited. Advances in pharmacodynamics have pointed out the importance of understanding the profiles of drugs in the plasma and in specialized compartments in order to dose the drugs to obtain the best patient outcomes. Advances in population pharmacokinetic modeling hold the promise of allowing proper estimation of drug penetration into the vitreous (or other specialized compartment) with only a single vitreous sample, in conjunction with plasma sampling. We have developed a rabbit model which allows multiple samples of vitreous to be obtained without breaking down the blood-vitreous barrier. We have employed this model to test the hypothesis that robust estimates of vitreous penetration by the fluoroquinolone ciprofloxacin can be obtained from a traditional intensive plasma sampling set plus a single vitreous sample. We studied 33 rabbits which were receiving 40 mg of ciprofloxacin per kg of body weight intravenously as short infusions and from which multiple plasma and vitreous samples were obtained and assayed for ciprofloxacin content by high performance liquid chromatography. Data were analyzed by the iterative two-stage population modeling technique (IT2S), employing the iterative two-stage program of Forrest et al. (Antimicrob. Agents Chemother. 37:1065-1072, 1993). Two data sets were analyzed: all plasma and vitreous samples versus all plasma samples and the initially obtained single vitreous sample. The pharmacokinetic parameter values identified were used to calculate the percent vitreous penetration as the ratio of the area under the concentration-time curve for the vitreous to that for the plasma. The values identified, 4% penetration for the full data set versus 3% penetration for the single vitreous sample data set, and their corresponding estimates were not statistically significantly different. We conclude that population modeling holds promise for the analysis of penetration of antimicrobiol agents into specialized spaces from which only single samples can be obtained, particularly for patients with whom robust plasma sampling can be performed. PMID- 7486902 TI - In vitro evaluation of combination of fluconazole and flucytosine against Cryptococcus neoformans var. neoformans. AB - Amphotericin B and fluconazole are current acceptable therapies for cryptococcal meningitis; however, their effect remains suboptimal. The combination of fluconazole and flucytosine has yielded encouraging clinical results in human immunodeficiency virus patients with cryptococcal meningitis. To investigate the biological basis of this finding, we performed in vitro combination testing of fluconazole and flucytosine against 50 clinical strains of Cryptococcus neoformans var. neoformans. Synergy (fractional inhibitory concentration index of < 1.0) was observed in 62% of cases, while antagonism (fractional inhibitory concentration index of > 2.0) was not observed. For cases in which synergy was not achieved (autonomous or additive effects), the beneficial effect of the combination was still seen (i.e., there was still a decrease, although not as dramatic, in the MIC of one or both drugs when used in combination). The in vitro inhibitory action of flucytosine was greatly enhanced by the addition of fluconazole; the flucytosine MICs for Cryptococcus isolates were markedly decreased to concentrations which were severalfold lower than the achievable cerebrospinal fluid flucytosine concentration. On the other hand, the addition of flucytosine did not greatly enhance the in vitro activity of fluconazole if the initial fluconazole MIC for the isolate was > or = 8 micrograms/ml. Controlled clinical studies are warranted to further elucidate the potential utility of fluconazole-flucytosine combination therapy. PMID- 7486903 TI - Fluconazole resistance due to energy-dependent drug efflux in Candida glabrata. AB - We report on the mechanism of fluconazole resistance in Candida glabrata from a case of infection in which pre- and posttreatment isolates were available for comparison. The resistant, posttreatment isolate was cross-resistant to ketoconazole and itraconazole, in common with other azole-resistant yeasts. Resistance was due to reduced levels of accumulation of [3H]fluconazole rather than to changes at the level of ergosterol biosynthesis. Studies with metabolic or respiratory inhibitors showed that this phenomenon was a consequence of energy dependent drug efflux, as opposed to a barrier to influx. Since energy-dependent efflux is a characteristic of multidrug resistance in bacteria, yeasts, and mammalian cells, we investigated the possibility that fluconazole resistance is mediated by a multidrug resistance-type mechanism. Benomyl, a substrate for the Candida albicans multidrug resistance protein, showed competition with fluconazole for efflux from resistance C. glabrata isolates, consistent with a common efflux mechanism for these compounds. By contrast, other standard substrates or inhibitors of multidrug resistance proteins had no effect on fluconazole efflux. In conclusion, we have identified energy-dependent efflux of fluconazole, possibly via a multidrug resistance-type transporter, as the mechanism of resistance to fluconazole in C. glabrata. PMID- 7486904 TI - Characterization of fluoroquinolone-resistant mutant strains of Mycobacterium tuberculosis selected in the laboratory and isolated from patients. AB - To examine the mechanism of resistance to fluoroquinolones in Mycobacterium tuberculosis, we selected spontaneous fluoroquinolone-resistant mutants from a susceptible strain, H37Rv, and studied the susceptibilities of these mutants and two fluoroquinolone-resistant clinical isolates (A-382, A-564) to various fluoroquinolones and to isoniazid and rifampin. Furthermore, since mutations within the quinolone resistance-determining region of the structural gene encoding the A subunit of DNA gyrase are the most common mechanism of acquired resistance, we amplified this region by PCR and compared the nucleotide sequences of the fluoroquinolone-resistant strains with that of the susceptible strain. Fluoroquinolone-resistant mutants of H37Rv appeared at frequencies of 2 x 10(-6) to 1 x 10(-8). For three mutants selected on ciprofloxacin, ofloxacin, and sparfloxacin, respectively, and the two clinical isolates, MICs of ciprofloxacin and ofloxacin were as high as 16 micrograms/ml, and those of sparfloxacin were 4 to 8 micrograms/ml. They displayed cross-resistance to all fluoroquinolones tested but not to isoniazid or rifampin. Sparfloxacin and FQ-A (PD 127391-0002) were the most potent fluoroquinolones. All of the fluoroquinolone-resistant strains (MICs, > or = 4 micrograms/ml) had mutations in the quinolone resistance determining region which led to substitution of the Asp residue at position 87 (Asp-87) by Asn or Ala or the substitution of Ala-83 by Val in the A subunit of DNA gyrase. Similar mutations have been noted in other bacterial species and recently in mycobacteria. The broad resistance to fluoroquinolones that arose readily by point mutation in the laboratory and apparently during inadequate therapy, as was the case in the clinical isolates, may ultimately lead to to serious restriction of the use of these drugs in the treatment of tuberculosis. PMID- 7486905 TI - Cross-resistance analysis of human immunodeficiency virus type 1 variants individually selected for resistance to five different protease inhibitors. AB - Human immunodeficiency virus type 1 (HIV-1) protease inhibitor-resistant variants, isolated on passage of HIV-1HXB2 in MT-4 cells with five different protease inhibitors, have been examined for cross-resistance to five inhibitors. The protease inhibitors studied were Ro 31-8959, A-77003, XM323, L-735,524, and VX-478. Resistant variants with two to four mutations within their protease sequence and 9- to 40-fold-decreased susceptibility were selected for all five inhibitors within six to eight passes in cell culture. Passage of a zidovudine resistant mutant in Ro 31-8959 generated a dual reverse transcriptase- and protease-resistant virus. Variants were cloned directly into a modified pHXB2-D infectious clone for cross-resistance analysis. Although the resistant variants selected possessed different combinations of protease mutations for each inhibitor, many showed cross-resistance to the other inhibitors, and one showed cross-resistance to all five inhibitors. Interestingly, some mutants showed increased susceptibility to some inhibitors. Further HIV passage studies in the combined presence of two protease inhibitors demonstrated that in vitro it was possible to delay significantly selection of mutations producing resistance to one or both inhibitors. These studies indicate that there may be some rationale for combining different protease inhibitors as well as protease and reverse transcriptase inhibitors in HIV combination therapy. PMID- 7486906 TI - In vitro pharmacodynamics of piperacillin, piperacillin-tazobactam, and ciprofloxacin alone and in combination against Staphylococcus aureus, Klebsiella pneumoniae, Enterobacter cloacae, and Pseudomonas aeruginosa. AB - The time-kill curve methodology was used to determine the pharmacodynamics of piperacillin, ciprofloxacin, piperacillin-tazobactam and the combinations piperacillin-ciprofloxacin and ciprofloxacin-piperacillin-tazobactam. Kill curve studies were performed for piperacillin, ciprofloxacin, and piperacillin tazobactam at concentrations of 0.25 to 50 times the MICs for 13 strains of bacteria: four Pseudomonas aeruginosa, three Enterobacter cloacae, three Klebsiella pneumoniae, and three Staphylococcus aureus isolates (tazobactam concentrations of 0.5, 4, and 12 micrograms/ml). By using a sigmoid Emax model and nonlinear least squares regression, the 50% lethal concentrations and the maximum lethal rates of each agent were determined for each bacterial strain. For piperacillin-ciprofloxacin and ciprofloxacin-piperacillin-tazobactam, kill curve studies were performed with concentrations obtained by the fractional maximal effect method (R. C. Li, J. J. Schentag, and D. E. Nix, Antimicrob. Agents Chemother. 37:523-531, 1993) and from individual 50% lethal concentrations and maximum lethal rates. Ciprofloxacin-piperacillin-tazobactam was evaluated only against the four P. aeruginosa strains. Interactions between piperacillin and ciprofloxacin were generally additive. At physiologically relevant concentrations of piperacillin and ciprofloxacin, ciprofloxacin had the highest rates of killing against K. pneumoniae. Piperacillin-tazobactam (12 micrograms/ml) had the highest rate of killing against E. cloacae. Piperacillin-ciprofloxacin with relatively higher ciprofloxacin concentrations had the greatest killing rates against S. aureus. This combination had significantly higher killing rates than piperacillin (P < 0.002). For all the bacterial strains tested, killing rates by ciprofloxacin were significantly higher than those by piperacillin-tazobactam (4 and 12 micrograms/ml had significantly higher killing rates than piperacillin alone (P < 0.02 and P < 0.004, respectively). The effect of the combination of piperacillin ciprofloxacin, in which piperacillin concentrations were relatively higher, was not statistically different from that of piperacillin alone (p > or = 0.71). The combination of ciprofloxacin-piperacillin-tazobactam achieved greater killing than other combinations or monotherapies against P. aeruginosa. The reduction in the initial inoculum was 1 to 4 logs greater with ciprofloxacin-piperacillin tazobactam at 4 and 12 micrograms/ml than with any other agent or combination of agents. On the basis of the additive effects prevalently demonstrated in the in vitro study, the combinations of piperacillin-ciprofloxacin and piperacillin tazobactam are rational therapeutic options. Greater killing of P. aeruginosa was demonstrated with ciprofloxacin-piperacillin--tazobactam. Since treatment failure of P. aeruginosa pneumonia is a significant problem, clinical studies are warranted. PMID- 7486907 TI - Partial characterization and effect of omeprazole on ATPase activity in Helicobacter pylori by using permeabilized cells. AB - ATPase activity in permeabilized cells of Helicobacter pylori as well as those of Helicobacter felis and Campylobacter jejuni was analyzed. The ATPase activities in these cells were most susceptible to sodium azide, fluoroaluminate, and dicyclohexylcarbodiimide, which are typical inhibitors of F ATPases. Optimal values for maximal activity were found to be at approximately pH 6.4, 6.0, and 6.0 for C. jejuni, H. pylori, and H. felis, respectively. The substituted benzimidazole compounds omeprazole, lansoprazole, and Eisai 3810 were found to have no effect on the F ATPase activity of H. pylori at concentrations which are inhibitory for cell growth (MICs). In addition, an extracellular, vanadate susceptible ATPase activity was detected in H. pylori, which was also relatively insusceptible to the benzimidazole compounds. Thus, the mechanism of killing mediated by omeprazole and related compounds in Helicobacter pylori does not appear to be due to diminished ATPase activity. PMID- 7486908 TI - Sequential, single-dose pharmacokinetic evaluation of meropenem in hospitalized infants and children. AB - Meropenem is a new carbapenem antibiotic which possesses a broad spectrum of antibacterial activity against many of the pathogens responsible for pediatric bacterial infections. In order to define meropenem dosing guidelines for children, an escalating, single-dose, pharmacokinetic study at 10, 20, and 40 mg/kg of body weight was performed. A total of 73 infants and children in four age groups were enrolled in the study: 2 to 5 months, 6 to 23 months, 2 to 5 years, and 6 to 12 years. The first patients enrolled were those in the oldest age group, who received the lowest dose. Subsequent enrollment was determined by decreasing age and increasing dose. Complete studies were performed on 63 patients. No age- or dose-dependent effects on pharmacokinetic parameter estimates were noted. Mean pharmacokinetic parameter estimates were as follows: half-life, 1.13 +/- 0.15 h; volume of distribution at steady state, 0.43 +/- 0.06 liters/kg; mean residence time, 1.57 +/- 0.11 h; clearance, 5.63 +/- 0.75 ml/min/kg; and renal clearance, 2.53 +/- 0.50 ml/min/liters kg. Approximately 55% of the administered dose was recovered as unchanged drug in the urine during the 12 h after dosing. No significant side effects were reported in any patients. By using the derived pharmacokinetic parameter estimates, a dose of 20 mg/kg given every 8 h will maintain plasma meropenem concentrations above the MIC that inhibits 90% of strains tested for virtually all potentially susceptible bacterial pathogens. PMID- 7486909 TI - New system based on site-directed mutagenesis for highly accurate comparison of resistance levels conferred by SHV beta-lactamases. AB - We developed a system based on site-directed mutagenesis that allows a precise comparison of SHV enzymes under isogenic conditions. In addition, the influences of two different, naturally occurring promoters were examined for each SHV derivative. The system comprised two separately cloned DNA fragments, each the size of 3.6 kb. Both fragments encoded an SHV gene originating from clinical isolates but with different promoters. The structural genes were made identical by site-directed mutagenesis. Other mutations were then introduced into both fragments by means of site-directed mutagenesis, resulting in the SHV derivatives SHV-1, SHV-2, SHV-2a, SHV-3, and SHV-5. The amino acid exchange of glutamic acid at position 235 for lysine in SHV-5 resulted in the highest resistance levels. SHV-3, differing from SHV-2 by the exchange of arginine at position 201 for leucine and previously described as indistinguishable from SHV-2, was shown to cause slightly higher resistance to ceftazidime and lower resistance to ceftriaxone, cefotaxime, and cefepime than SHV-2. The point mutation in SHV-2a, with the leucine-to-glutamine replacement at the unusual position 31, previously considered almost insignificant, proved to increase resistance to ceftazidime but reduced the MICs of all other cephalosporins tested when compared with those for SHV-2. For all clones harboring SHV derivatives, resistance was increased by a stronger promoter, in some cases masking the effect of the point mutation itself and demonstrating the importance of regulatory mechanisms of resistance. PMID- 7486910 TI - Purification of a 54-kilodalton protein (OprJ) produced in NfxB mutants of Pseudomonas aeruginosa and production of a monoclonal antibody specific to OprJ. AB - The 54-kDa outer membrane protein (designated OprJ) of a norfloxacin-resistant nfxB mutant of Pseudomonas aeruginosa PAO1 was purified by ion-exchange high performance liquid chromatography. Mobility of OprJ in sodium dodecyl sulfate polyacrylamide gel electrophoresis was not affected by reduction and heating. A murine monoclonal antibody (MAb) against OprJ was prepared to investigate existence of this protein in fluoroquinolone-susceptible and -resistant strains of P. aeruginosa. Western blot (immunoblot) analysis with this MAb revealed a single band at the position corresponding to OprJ in outer membrane proteins of NfxB mutants derived from clinical isolates. However, the MAb did not react with any outer membrane proteins of the respective parent strains. Complementation of the NfxB mutation by transformation with plasmid pNF111, which contained the wild type nfxB gene, led to disappearance of the single band corresponding to OprJ. The existence of OprJ was associated with fluoroquinolone resistance. Furthermore, the MAb did not react with any outer membrane proteins of other fluoroquinolone-resistant nalB and nfxC mutants. These results suggest that OprJ is newly produced in NfxB mutants of P. aeruginosa and is involved in fluoroquinolone resistance specific to NfxB, and it appears that the MAb to OprJ should aid in detection of the NfxB mutation in P. aeruginosa. PMID- 7486911 TI - In vivo activity and pharmacodynamics of cefotaxime or ceftriaxone in combination with fosfomycin in fibrin clots infected with highly penicillin-resistant Streptococcus pneumoniae. AB - Using a clinical pneumococcal strain for which MICs were 2, 0.5, 0.5, and 16 mg/liter for penicillin, cefotaxime, ceftriaxone, and fosfomycin, respectively, we studied the efficacies of these antibiotics alone and in combination in one or two doses or in continuous infusion over 6 h in the treatment of the prolonged (48-h) experimental fibrin clot infections in rabbits. Doses were chosen to obtain low antibiotic concentrations. We observed the highest bacterial reductions (change in log10 CFU per gram) with the following five regimens: combination of cefotaxime plus fosfomycin given in two divided doses 6 h apart (each at 50 mg/kg of body weight given intravenously (4.2 +/- 0.7 CFU/g), ceftriaxone (8 mg/kg given once intravenously) along with one or two doses of fosfomycin (3.79 +/- 0.6 and 3.95 +/- 0.5 CFU/g), cefotaxime alone administered in two divided doses (3.6 +/- 0.4 CFU/g), and a 6-h continuous infusion of cefotaxime (100 mg/kg) with fosfomycin (100 mg/kg) (3.5 +/- 0.4 CFU/g). The bacterial reductions obtained with these five regimens were all higher than those obtained with the other regimens tested (P < 0.05). The time of bacterial regrowth was significantly delayed with the two doses of the cefotaxime fosfomycin regimens (23.2 +/- 11 h) compared with those with the other combinations (P < 0.05). The rate of bacterial regrowth with this regimen was even lower than that observed with cefotaxime alone given in two doses (P < 0.05). By a multivariate analysis, the most important independent parameters for efficacy were the maximal concentrations of beta-lactam antibiotics and the residual concentration of fosfomycin and, for the combinations, the log of the area under the concentration-time curve/MIC ratio for beta-lactam antibiotics. From these findings, the combinations cefotaxime or ceftriaxone plus fosfomycin could be proposed for the treatment of infections caused by highly penicillin resistant pneumococci. PMID- 7486912 TI - Interleukin-1 (IL-1)-induced resistance to bacterial infection: role of the type I IL-1 receptor. AB - Pretreatment with a low dose of recombinant human interleukin-1 beta (IL-1) (3 to 30 micrograms/kg) 24 h before a lethal Pseudomonas aeruginosa infection prolongs survival in neutropenic mice. We investigated the role of the type I IL-1 receptor (IL-1RI) and IL-1RII in this IL-1-induced protection by using a specific IL-1 receptor antagonist (IL-1-Ra), which blocks effects mainly via IL-1RI. Pretreatment with IL-1Ra before IL-1 partially blocked the IL-1-induced enhanced survival, whereas pretreatment with a specific neutralizing monoclonal antibody to IL-1RI (35F5) eliminated the IL-1 induced protection. The nonapeptide fragment 163-171 of recombinant human IL-1 beta, which possesses the immunoadjuvant but not the inflammatory effect of the entire molecule via a non-receptor-mediated signal transduction process, did not reproduce the IL-1-induced protection. IL-1 induced protection was associated with reduced serum aspartate aminotransferase and alanine aminotransferase concentrations in conjunction with ameliorated histopathology of the liver. These findings may be due to reduced cytokine production and cytokine sensitivity of target cells after infection. We conclude that the IL-1-induced nonspecific resistance to infection is mediated by cells bearing IL-1RI and is associated with a reduction of liver damage. PMID- 7486913 TI - Influence of aspirin on development and treatment of experimental Staphylococcus aureus endocarditis. AB - Previously, we have shown that a 5-mg/kg of body weight daily dose of aspirin (ASA) caused reductions in the bacterial densities and weights of aortic vegetations in a rabbit model of Staphylococcus aureus endocarditis. We sought to determine (i) whether ASA dosage influences the development of vegetations and (ii) whether ASA given with antimicrobial therapy improves the treatment outcome of infective endocarditis. To study the influence of ASA dosage, animals received either no ASA (control) or oral doses of 2.5, 10, 20, and 50 mg/kg daily. The 2.5 and 10-mg/kg groups had statistically significant reductions in vegetation weight compared with untreated controls. The 10-mg/kg dose also resulted in a significant decrease in bacterial densities compared with those of the controls. Although reductions in weight and bacterial density were observed in other ASA treated groups, these did not achieve statistical significance. To study the influence of ASA and antimicrobial therapy, the animals received either vancomycin alone or vancomycin with ASA. When ASA was given prior to and during antimicrobial therapy, a significant reduction in vegetation weight was observed. Additionally, the rate of sterilization was directly proportional to this observed reduction in weight. ASA's impact on the reduction of both the bacterial density and the weight of aortic vegetations is a dose-dependent phenomenon. When given with antimicrobial therapy, ASA not only reduces vegetation weight but also improves the rate of sterilization. This study provides additional data regarding the role of ASA in the treatment of endocarditis. PMID- 7486914 TI - Lipid-based slow-release formulation of amikacin sulfate reduces foreign body associated infections in mice. AB - Treatment and prophylaxis of uncomplicated infections with standard systemic antibiotics are usually successful. However, standard systemic antibiotic therapy alone is frequently unsatisfactory in certain circumstances, such as the presence of a foreign body (FB), necrotic tissue, overwhelming bacterial inoculum, or poor vascular supply to the involved tissues. We have developed a lipid-based sustained release formulation of amikacin sulfate (DepoFoam encapsulated amikacin sulfate [DEAS]) as a biodegradable, locally injectable antibiotic for such circumstances. The encapsulated drug is released over 7 to 10 days. We tested the efficacy of this formulation in an FB infection model in which Teflon tubes (length, 1 cm; outside diameter, 1.6 mm) were implanted into the subcutaneous tissue in mice and the local site was inoculated with 0.87 x 10(7) CFU of Staphylococcus aureus 3 days later. Inoculation was followed by either no treatment or a local injection of DEAS, free amikacin sulfate, non-drug containing DepoFoam, or systemic free amikacin sulfate. All drug applications contained 1 mg of amikacin. One group was implanted with the FB and left unchallenged with bacteria and untreated as a sterile control group. All animals were sacrificed 10 days following FB implantation. FBs were retrieved from tissues by an aseptic technique and incubated in liquid culture media for 7 days. Local wound tissue was excised and processed to determine the number of CFU per gram of tissue. Treatment with local or systemic free amikacin had no effect on the number of infected FBs or on the log CFU in wound tissue compared with the untreated or non-drug-containing DepoFoam group. Compared with local free amikacin therapy, the number of infected FBs was reduced from 86 to 25% (P=0.02) following treatment with DEAS, and log CFU per gram of tissue was significantly decreased from 4.8 +/- 0.9 to 1.3 +/- 0.6 (P<0.0005). DEAS may have clinical utility as locally injected antibiotic in certain infections. PMID- 7486915 TI - Inhibition of recombinant Pneumocystis carinii dihydropteroate synthetase by sulfa drugs. AB - Forty-four sulfa drugs were screened against crude preparations of recombinant Pneumocystis carinii dihydropteroate synthetase. The apparent Michaelis-Menten constants (Km) for p-aminobenzoic acid and 7,8-dihydro-6-hydroxymethylpterin pyrophosphate were 0.34 +/- 0.02 and 2.50 +/- 0.71 microM, respectively. Several sulfa drugs, including sulfathiazole, sulfachlorpyridazine, sulfamethoxypyridazine, and sulfathiourea, inhibited dihydropteroate synthetase approximately as well as sulfamethoxazole, as determined by the concentrations which cause 50% inhibition and/or by Ki. For all sulfones and sulfonamides tested, unsubstituted p-amino groups were necessary for activity, and sulfonamides containing an N1-heterocyclic substituent were found to be the most effective inhibitors. Folate biosynthesis in isolated intact P. carinii was approximately equally sensitive to inhibition by sulfamethoxazole, sulfachlorpyridazine, sulfamethoxypyridazine, sulfisoxazole, and sulfathiazole. Two of these drugs, sulfamethoxypyridazine and sulfisoxazole, are known to be less toxic than sulfamethoxazole and should be further evaluated for the treatment of P. carinii pneumonia. PMID- 7486916 TI - Bactericidal killing activities of cefepime, ceftazidime, cefotaxime, and ceftriaxone against Staphylococcus aureus and beta-lactamase-producing strains of Enterobacter aerogenes and Klebsiella pneumoniae in an in vitro infection model. AB - Cefepime (CP) is a new injectable cephalosporin with a broad spectrum of activity and stability against common chromosomally and plasmid-mediated beta-lactamases. The bactericidal activities of CP, ceftazidime (CZ), cefotaxime (CTX), and ceftriaxone (CAX) against reference and clinical strains of Staphylococcus aureus, an isogenic pair of Enterobacter aerogenes strains (wild type and a CZ resistant derepressed mutant), and a Klebsiella pneumoniae isolate possessing a TEM-10 beta-lactamase were investigated in a two-compartment pharmacodynamic in vitro infection model which simulates human pharmacokinetics. An inoculum of approximately 10(6) CFU/ml was used in all model experiments. Antibiotics were administered to simulate the following regimens: CP at 2 g every 12 h (q12h), CZ at 2 g q8h, CTX at 2 g q8h, and CAX at 2 g q24h. Human albumin was added during experiments with CAX and staphylococci to simulate protein binding. Samples were removed at multiple time points over a 48-h period to determine the inoculum size for time-kill curves. Development of resistance was detected by inoculating samples obtained at 0, 24, and 48 h onto antibiotic-containing agar plates. The time to 99.9% killing was used to compare drug regimens. Against staphylococci, the time to bacterial eradication was significantly delayed with CAX-albumin. All regimens had similar activities against the wild-type Enterobacter strain; however, regrowth was noted with CZ, CTX, and CAX against the CZ-resistant strain. There were no differences between the CP, CTX, and CAX regimens against K. pneumoniae. Of interest, no regrowth of any organism was noted with CP. These data indicate that CP has activity against S.aureus and CZ-resistant gram negative bacilli. PMID- 7486917 TI - Use of primers selective for vancomycin resistance genes to determine van genotype in enterococci and to study gene organization in VanA isolates. AB - Vancomycin resistance in enterococci is an emerging therapeutic problem. Resistance is not always detected by standard microbiological methods. Oligonucleotide primers for PCR were designed to target amplification of defined regions of genes of the vanA cluster, as well as vanB and vanC1. These primers correctly identified 30 vancomycin-resistant isolates tested (17 VanA, 7 VanB, and 6 Enterococcus gallinarum). No amplification was observed with Enterococcus casseliflavus or vancomycin-susceptible strains. Using PCR and Southern blotting, we found that all 17 VanA isolates had orf-1, orf-2, vanR, vanS, vanH, vanA, and vanY genes in the same sequence and that the intergenic distances in the vanR vanA segments were the same. The described methods should be applicable to the rapid detection of the different vancomycin resistance genotypes in enterococci. PMID- 7486918 TI - Antagonistic interactions between azoles and amphotericin B with yeasts depend on azole lipophilia for special test conditions in vitro. AB - The interactions of the azole antifungal agents fluconazole, itraconazole, ketoconazole, or miconazole with amphotericin B (AmB) in their effect on Candida albicans were investigated. These four azoles antagonized the fungistatic activity of AmB at sub-MICs if both substances acted simultaneously. This coincubation test was primarily developed to observe the azole-mediated demethylase inhibition quantitatively by bioassay. Interestingly, the occurrence of azole-AmB antagonism depended on azole lipophilia if specially selected test conditions were applied. By a consecutive incubation regimen, preincubation at high azole concentrations (1 to 50 micrograms/ml) and then subsequent incubation with AmB (1 microgram/ml), only preincubation with the three lipophilic azoles decreased the fungicidal activity of AmB but not that of FCZ. It was shown that yeasts absorb only lipophilic azoles to a remarkable extent. This fact might be responsible for the absence of antagonism of FCZ to AmB when yeasts were incubated consecutively. It can be concluded with caution that consecutive treatment of candidiasis with FCZ and AmB does not necessarily result in a clinically relevant antagonism. PMID- 7486919 TI - Aerosol and parenteral pneumocandins are effective in a rat model of pulmonary aspergillosis. AB - The pneumocandins are semisynthetic analogs of echinocandin-like compounds that have shown efficacy in animal models of systemic candidiasis, disseminated aspergillosis, and pneumocystis pneumonia. However, the most common form of Aspergillus infection in susceptible patients is pulmonary aspergillosis, which was not directly tested in the mouse models used in the past. We have evaluated three pneumocandins, L-693,989, L-731,373, and L-733,560, in a rat model of pulmonary aspergillosis. Male Sprague-Dawley rats were treated for 2 weeks with cortisone and tetracycline and fed a low-protein diet before being inoculated via the trachea with 10(6) conidia of Aspergillus fumigatus H11-20. In the absence of drug treatment, the animals developed a progressive, rapidly fatal bronchopneumonia. All three pneumocandins at doses of 5 mg/kg (intraperitoneally [i.p.] every 12 h [q12h]) were effective in delaying mortality in this model. Survival at day 7 postinfection was 20% among controls (n = 10 for all groups), while it was 60, 80, and 90% in groups that were treated with L-693,989, L 731,373, and L-733,560, respectively. In another trial, survival at day 7 postinfection was 25% among controls (n = 8 for all groups); it was 87.5% in a group treated with amphotericin B (0.5 mg/kg i.p. q12h) and was 100% in a group treated with L-733,560 (0.625 mg/kg i.p. q12h). In a separate trial, aerosol L 693,989 administered 2 h before infection (5 mg/kg) delayed mortality. Eight of the 10 animals treated with aerosol L-693,989 survived for 7 days, whereas only 2 of 10 control animals survived. We conclude that the pneumocandins we tested were highly effective in an animal model of pulmonary aspergillosis. PMID- 7486921 TI - Pharmacodynamics of ceftazidime administered as continuous infusion or intermittent bolus alone and in combination with single daily-dose amikacin against Pseudomonas aeruginosa in an in vitro infection model. AB - We compared the pharmacodynamics and killing activity of ceftazidime, administered by continuous infusion and intermittent bolus, against Pseudomonas aeruginosa ATCC 27853 and ceftazidime-resistant P. aeruginosa 27853CR with and without a single daily dose of amikacin in an in vitro infection model over a 48 h period. Resistance to ceftazidime was selected for by serial passage of P. aeruginosa onto agar containing increasing concentrations of ceftazidime. Human pharmacokinetics and dosages were simulated as follows: half-life, 2 h; intermittent-bolus ceftazidime, 2 g every 8 h (q8h) and q12h; continuous infusion, 2-g loading dose and maintenance infusions of 5, 10, and 20 micrograms/ml; amikacin, 15 mg/kg q24h. There was no significant difference in time to 99.9% killing between any of the monotherapy regimens or between any combination regimen against ceftazidime-susceptible P. aeruginosa. Continuous infusions of 10 and 20 micrograms/ml killed as effectively as an intermittent bolus of 2 g q12h and q8h, respectively. Continuous infusion of 20 micrograms/ml and an intermittent bolus of 2 g q8h were the only regimens which prevented organism regrowth at 48 h, while a continuous infusion of 5 micrograms/ml resulted in the most regrowth. All of the combination regimens exhibited a synergistic response, with rapid killing of ceftazidime-susceptible P. aeruginosa and no regrowth. Against ceftazidime-resistant P. aeruginosa, none of the ceftazidime monotherapy regimens achieved 99.9% killing. The combination regimens exhibited the same rapid killing of the resistant strain as occurred with the susceptible strain; however, regrowth occurred with all regimens. The combination regimens of continuous infusion of 20 micrograms/ml plus amikacin and intermittent bolus q8h or q12h plus amikacin continued to be synergistic. Overall, continuous infusion monotherapy with ceftazidime at concentrations 4 to 5 and 10 to 15 times the MIC was as effective as an intermittent bolus of 2 g q12h (10 to 15 times the MIC) and q8h (25 to 35 times the MIC), respectively, against ceftazidime-susceptible P. aeruginosa. Combination therapy with amikacin plus ceftazidime, either intermittently q8h or by continuous infusion of 20 micrograms/ml, appeared to be effective and exhibited synergism against ceftazidime-resistant P. aeruginosa. PMID- 7486920 TI - Cloning and characterization of a 3-N-aminoglycoside acetyltransferase gene, aac(3)-Ib, from Pseudomonas aeruginosa. AB - A novel gene encoding an aminoglycoside 3-N-acetyltransferase, which confers resistance to gentamicin, astromicin, and sisomicin, was cloned from Pseudomonas aeruginosa Stone 130. Its sequence was determined and found to show considerable similarity to an aac(3)-I gene previously cloned from R plasmids from Enterobacter, Pseudomonas, and Serratia spp. We have designated the genes from the R plasmids and this work aac(3)-Ia and aac(3)-Ib, respectively. The two aac(3)-I genes share 74% nucleotide identity, and their deduced protein products are 88% similar. These data suggest that the genes derive from a common ancestor. Homology between the flanking sequences of both aac(3)-I genes and other resistance determinants known to reside in integron environments was also observed. Intragenic probes specific for either aac(3)-Ia or aac(3)-Ib were used in hybridization studies with a series of gentamicin-, astromicin-, and sisomicin resistant clinical isolates. Of 59 clinical isolates tested, no isolates hybridized with both probes, 30 (51%) hybridized with the aac(3)-Ia probe, 12 (20%) hybridized with the aac(3)-Ib probe, and 17 (29%) did not hybridize with either probe. These data suggest the existence of at least one other aac(3)-I gene. PMID- 7486922 TI - Mode of action of (R)-9-[4-hydroxy-2-(hydroxymethyl)butyl]guanine against herpesviruses. AB - The activity, metabolism, and mode of action of (R)-9-[4-hydroxy-2 (hydroxymethyl)butyl]guanine (H2G) against herpes simplex virus type 1 (HSV-1) and type 2 (HSV-2) and varicella-zoster virus (VZV) were studied. Compared to acyclovir (ACV), H2G has superior activity against VZV (50% inhibitory concentration of 2.3 microM) and Epstein-Barr virus (50% inhibitory concentration of 0.9 microM), comparable activity against HSV-1, and weaker activity against HSV-2. The antiviral effect on HSV-1 showed persistence after removal of compound. H2G was metabolized to its mono-, di- and triphosphate derivatives in virus-infected cells, with H2G-triphosphate being the predominant product. Only small amounts of H2G-triphosphate were detected in uninfected cells (1 to 10 pmol/10(6) cells), whereas the level in HSV-1-infected cells reached 1,900 pmol/10(6) cells. H2G was a substrate for all three viral thymidine kinases and could also be phosphorylated by mitochondrial deoxyguanosine kinase. The intracellular half-life of H2G-triphosphate varied in uninfected (2.5 h) and infected (HSV-1, 14 h; VZV, 3.7 h) cells but was always longer than the half-life of ACV-triphosphate (1 to 2 h). H2G-triphosphate inhibited HSV-1, HSV-2, and VZV DNA polymerases competitively with dGTP (Ki of 2.8, 2.2, and 0.3 microM, respectively) but could not replace dGTP as a substrate in a polymerase assay. H2G was not an obligate chain terminator but would only support limited DNA chain extension. Only very small amounts of radioactivity, which were too low to be identified by high-performance liquid chromatography analysis of the digested DNA, could be detected in purified DNA from uninfected cells incubated with [3H]H2G. Thus, H2G acts as an anti-herpesvirus agent, particularly potent against VZV, by formation of high concentrations of relatively stable H2G-triphosphate, which is a potent inhibitor of the viral DNA polymerases. PMID- 7486923 TI - Efficacy of D0870 compared with those of itraconazole and amphotericin B in two murine models of invasive aspergillosis. AB - D0870 is a novel azole antifungal compound. It was compared with conventional amphotericin B and itraconazole therapy in two murine models of invasive aspergillosis, one a systemic nonimmunocompromised mouse model and the other a temporarily neutropenic mouse respiratory model. D0870 was given orally and achieved measurable concentrations in serum approximately proportional to the daily dose with accumulation over time if it was given twice daily. Amphotericin B at 3.3 mg/kg of body weight was given intraperitoneally for four to six doses, and itraconazole was given orally in a cyclodextrin suspension at 5 to 50 mg/kg daily or twice daily (BID). The duration of therapy varied from 7 to 14 days. In the nonimmunocompromised mouse model, D0870 at 25 mg/kg BID was slightly inferior to amphotericin B and itraconazole with regard to mortality, with a median survival of 20 days for the three groups (P = 0.03 compared with amphotericin B). However, D0870 at 25 mg/kg BID was inferior to amphotericin B (but not itraconazole) with respect to renal culture (P = 0.01) and brain culture (P = 0.0001) results. Only amphotericin B was statistically superior to controls with regard to mortality. In the neutropenic mouse respiratory model, D0870 at 50 mg/kg/day was superior to amphotericin B, itraconazole, and controls with regard to mortality. D0870 at both 25 and 50 mg/kg/day was statistically superior to controls with regard to lung culture results (P = 0.004 to 0.04). A second experiment with a higher inoculum showed that no drug regimen was effective in that model. In all models low doses and concentrations of D0870 in serum were ineffective. D0870 has some efficacy for the treatment of invasive aspergillosis when it is given at modest doses. PMID- 7486924 TI - Effective treatment of cephalosporin-rifampin combinations against cryptic methicillin-resistant beta-lactamase-producing coagulase-negative staphylococcal experimental endocarditis. AB - The efficacy of cefazolin or cefpirome alone or combined with rifampin was compared with that of vancomycin alone or combined with rifampin in an experimental model of methicillin-resistant, beta-lactamase-producing, coagulase negative staphylococcal endocarditis. Phenotypically, the mecA gene-positive strain used in vivo did not exhibit methicillin resistance by the agar dilution or disk susceptibility method but was resistant in vitro (oxacillin MIC, 64 micrograms/ml) by the microtiter dilution method with 2% NaCl supplementation. Macrodilution broth susceptibilities of standard inocula failed to demonstrate cross-resistance of staphylococci to cefazolin (MIC, 8 micrograms/ml) or cefpirome (MIC, 4 micrograms/ml). In vivo, vancomycin and cefpirome had similar activities, and both regimens were more effective than was cefazolin alone. While the MIC of rifampin was low (0.031 micrograms/ml), monotherapy with rifampin resulted in a bimodal distribution of outcomes due to the expected emergence of resistant mutants. The results in vitro of time-kill synergy studies using rifampin in combination with cefazolin or cefpirome varied with the antimicrobial concentrations tested and did not reliably predict activities in vivo of rifampin beta-lactam combination therapies. Cefpirome, but not cefazolin or vancomycin, in combination with rifampin was synergistic in vivo. Cefpirome in combination with rifampin was more effective than was cefazolin in combination with rifampin. Both cephalosporin-rifampin regimens were significantly more effective than was cephalosporin or vancomycin monotherapy and were as effective as vancomycin combined with rifampin. These data support further evaluation of rifampin-beta lactam combinations as possible alternative therapies to vancomycin-containing regimens for selected methicillin-resistant coagulase-negative staphylococcal infections. PMID- 7486925 TI - Effect of xylitol on growth of nasopharyngeal bacteria in vitro. AB - Xylitol is known to reduce caries by inhibiting the growth of Streptococcus mutans. We hypothesized that xylitol could also affect the growth of other nasopharyngeal bacterial flora, which could be important when considering respiratory infections caused by these bacteria. We studied this in vitro by adding xylitol to the medium and observed that 1 and 5% xylitol reduced markedly the growth of alpha-hemolytic streptococci, including S. pneumoniae. It reduced slightly the growth of beta-hemolytic streptococci but not that of Haemophilus influenzae or Moraxella catarrhalis. The inhibitory growth pattern was similar to that previously seen with S. mutans, which may indicate a similarity in the enzymatic processing of five-carbon sugars such as xylitol. This sugar alcohol is a widely used sweetener, and the concentrations used in our experiments are easily achieved in the oral cavity. If xylitol reduces the growth of S. pneumoniae in the nasopharynx, it could also reduce the carriage of this pathogen and thus have clinical significance in the prevention of pneumococcal diseases. PMID- 7486926 TI - Inhibition of growth of Histoplasma capsulatum yeast cells in human macrophages by the iron chelator VUF 8514 and comparison of VUF 8514 with deferoxamine. AB - Histoplasma capsulatum requires intracellular iron to survive and multiply within human and murine macrophages (M phi). Thus, iron chelators may be useful compounds in the treatment of histoplasmosis. In the present study we compared the efficacies of five different iron chelators with deferoxamine (DEF) for their capacity to inhibit the growth of H. capsulatum yeast cells in culture medium and within human M phi. Of the agents tested, only one, VUF 8514, a 2,2'-bipyridyl analog, was found to be effective. VUF 8514 inhibited the growth of yeast cells in tissue culture medium and within M phi in a dose-response fashion. In tissue culture medium, the 50% effective dose (ED50) of VUF 8514 was 30 nM and the ED50 of DEF was 1 mM. In human M phi, the ED50 of VUF 8514 was 520 nM and the ED50 of DEF was 4 mM. Thus, VUF 8514 was effective at a concentration 7.7 x 10(3)-fold lower than DEF in inhibiting the growth of yeast cells in M phi. Inhibition of the intracellular growth of yeast cells by VUF 8514 was reversed by holotransferrin and iron nitriloacetate, an iron compound that is soluble at neutral to alkaline pH. Thus, VUF 8514 inhibits the intracellular growth of yeast cells by acting as an iron chelator rather than through its capacity as a weak base. These data suggest that the hydroxamic acid siderophore of H. capsulatum yeast cells competes successfully for iron against some iron chelators but not others and that VUF 8514 may be a potential therapeutic agent for the treatment of histoplasmosis. PMID- 7486927 TI - The closely related ermB-ermAM genes from Clostridium perfringens, Enterococcus faecalis (pAM beta 1), and Streptococcus agalactiae (pIP501) are flanked by variants of a directly repeated sequence. AB - The Clostridium perfringens macrolide-lincosamide-streptogramin B resistance gene, ermBP, was sequenced and shown to be identical to the ermB-ermAM gene from the promiscuous Enterococcus faecalis plasmid pAM beta 1 and to have at least 98% nucleotide sequence identity to other ermB-ermAM genes. Flanking the ermBP structural gene were almost identical directly repeated 1,341-bp sequences (DR1 and DR2). These repeats potentially encoded a 298 (or 284)-amino-acid protein that had sequence similarity to chromosomal and plasmid partitioning proteins. The pAM beta 1 and Streptococcus agalactiae (pIP501) erm determinants appeared to have DR2 but had similar internal 973- or 956-bp deletions in DR1, respectively. Some of the other ermB-ermAM class determinants had small portions of DR1, but none had complete copies. It is postulated that the C. perfringens ermBP determinant was derived from an enterococcal or streptococcal determinant that had complete copies of both DR1 and DR2. PMID- 7486928 TI - Pharmacokinetics and bioavailability of fluconazole in two groups of males with human immunodeficiency virus (HIV) infection compared with those in a group of males without HIV infection. AB - Fluconazole pharmacokinetics, including absolute bioavailability, were determined for one group of controls (n = 10) and two groups of people with human immunodeficiency virus (HIV) infection (those with CD4+ T-cell counts of less than [n = 4] or greater than [n = 9] 200 cells per mm3). Twenty subjects received four doses of fluconazole; three doses were oral (50, 100, and 400 mg), and one dose was intravenous (either 50, 100, or 400 mg). The other three subjects received one or two doses. The groups were comparable in terms of the weight, body mass index, and estimated creatinine clearance of the subjects, but the people with HIV infection were older. Pharmacokinetic parameters indicated linearity in all subjects; the area under the plasma concentration-time curve and the maximum concentration increased in proportion to the dose. The fraction of an oral dose of fluconazole absorbed approximated unity in all three groups of subjects. The mean (+/- standard deviation) plasma clearance of fluconazole was lowest in the group of subjects with low CD4+ T-cell counts; the value for this group was 0.74 +/- 0.19 liter/h, compared with 0.97 +/- 0.19 liter/h in the group with HIV infection and CD4+ T-cell counts of greater than 200 cells/mm3 and 1.18 +/- 0.23 liter/h in the group of control subjects (P < 0.05). The volume of distribution was lower in those with HIV infection (P = 0.04, corrected for weight). The half-life was longest in people with HIV infection and low CD4+ T cell counts (P = 0.01). This study has shown that some differences do exist between the pharmacokinetics of fluconazole in people with HIV infection and those in noninfected controls. PMID- 7486929 TI - Protein binding and serum bactericidal activities of vancomycin and teicoplanin. AB - In a randomized crossover study, the protein binding and serum bactericidal activities (SBAs) of vancomycin and teicoplanin against Staphylococcus aureus and Streptococcus pyogenes were investigated in six healthy volunteers. Total concentrations in serum 1 h postadministration of vancomycin and teicoplanin were 25.5 +/- 2.7 and 10.8 +/- 8.9 mg/liter, respectively; mean free concentrations were 14.6 +/- 2.0 and 0.6 +/- 0.9 mg/liter, respectively. Protein binding for vancomycin was 36.9% +/- 2.87%, and that for teicoplanin was 97.4% +/- 2.6%. SBA determined in pooled human serum at 1 h against S. aureus ranged from 1:8 to 1:32 for both vancomycin and teicoplanin. Against S. pyogenes SBA at 1 h ranged from 1:16 to 1:128 for vancomycin and 1:256 to 1:2,048 for teicoplanin. In vitro kill curve studies showed that vancomycin is slowly bactericidal and that teicoplanin is bacteriostatic. Despite having less in vitro cidal activity against the study isolates and having low or unrecordable levels of free drug in serum, teicoplanin demonstrated a similar or better SBA than vancomycin. SBA was more closely related to the total drug level (r = 0.77 for S. aureus and r = 0.79 for S. pyogenes) than the free level of teicoplanin (r = 0.59 for S. aureus and r = 0.56 for S. pyogenes). The high level of protein binding of teicoplanin did not seem to impair its antibacterial activity as measured by its SBA. PMID- 7486930 TI - Pharmacodynamics of metronidazole determined by a time-kill assay for Trichomonas vaginalis. AB - The pharmacodynamic effects of metronidazole on Trichomonas vaginalis have been poorly characterized. The present in vitro study was performed to characterize the relationship between killing of trichomonads and metronidazole exposure (metronidazole concentration and time of exposure). Five laboratory strains and five recent clinical isolates of T. vaginalis were studied. The minimum lethal concentrations (MLCs) of metronidazole for the strains ranged from 0.8 to 25 micrograms/ml under anaerobic conditions. Metronidazole exhibited concentration dependent killing against T. vaginalis at concentrations ranging from 0.1 to > 10 times the MLC. The endpoint measurement, the kill rate constant, which was derived from the reduction in the logarithm of the colony count divided by exposure time, compared with the kill rate constant for the growth control was not affected by the time of assessment between 2 and 24 h. The kill rate constant versus-metronidazole exposure curves were similar when concentration was expressed as a multiple of the MLC. There were no apparent differences between the clinical isolates and laboratory strains. These data suggest that peak metronidazole concentration and/or area under the plasma concentration-versus time curve are the important pharmacodynamic parameters to be optimized. PMID- 7486931 TI - Interactions of Yersinia pestis penicillin-binding proteins with beta-lactam antibiotics. AB - The affinities of six major penicillin-binding proteins (PBPs) of Yersinia pestis EV76 to different beta-lactam antibiotics were determined. The results indicate that, similar to their counterparts in Escherichia coli, PBP2 and PBP3 are the lethal targets of amdinocillin and furazlocillin, respectively. The PBP contents of four additional Y. pestis strains and the morphological effects produced by some beta-lactam antibiotics are also reported. PMID- 7486932 TI - Effect of pH on activities of novel beta-lactamases and beta-lactamase inhibitors against these beta-lactamases. AB - The effects of acidic conditions on activities of seven beta-lactamases--TEM-1 (class A), KOXY (class A), IMP-1 (class B), AmpC (class C), MOX-1 (class C), OXA 5 (class D), and PSE-2 (class D)--and their inhibitors were measured. The enzymatic activities of KOXY, IMP-1, and MOX-1 at pH 5.8 were slightly lower than those at pH 7.5. However, the activities of PSE-2 and OXA-5 were greatly reduced at pH 5.8. All of the beta-lactamase inhibitors tested had poorer inhibitory activities at pH 5.8 than at pH 7.5 except clavulanic acid for TEM-1. PMID- 7486933 TI - Bactericidal and morphological effects of amoxicillin on Helicobacter pylori. AB - The growth kinetics of Helicobacter pylori after it has been exposed to amoxicillin have been investigated in conjunction with studies of cell morphology. A potent bactericidal effect was observed at concentrations 10-fold higher than the MIC, but this was accompanied by an increase in the residual numbers of coccoid forms observed. In the presence of 10 micrograms of amoxicillin per ml, these forms could be detected as rapidly as 6 h after exposure to the antibiotic. Although the clinical relevance of coccoid forms remains unknown, such forms should be considered when potential anti-Helicobacter agents are tested in vitro. PMID- 7486934 TI - In vitro activities of PS-15, a new dihydrofolate reductase inhibitor, and its cyclic metabolite against Mycobacterium avium complex. AB - PS-15, a new dihydrofolate reductase inhibitor, and its cyclic metabolite were evaluated for in vitro activity against 31 clinical Mycobacterium avium complex isolates. Broth dilution MICs of PS-15 ranged from 16 to 64 micrograms/ml. The cyclic metabolite was three to five times more active than the parent compound. Further evaluation of these compounds in an M. avium-infected murine test system will be of interest. PMID- 7486935 TI - Susceptibilities of Neisseria gonorrhoeae to the glycylcyclines. AB - To assess the activities of two glycylcyclines, N,N-dimethylglycylamido (DMG) derivatives of minocycline (MINO) and 6-demethyl-6-deoxytetracycline (DMDOT), 203 gonococcal isolates recovered at six sexually transmitted disease clinics in the western United States were evaluated. Antimicrobial susceptibilities to tetracycline HCl, doxycycline, MINO, DMG-DMDOT, and DMG-MINO were determined by agar dilution tests. DMG-DMDOT and DMG-MINO were more active than tetracycline HCl, doxycycline, or MINO regardless of the presence of Tet M or of chromosomal mutations mediating tetracycline resistance (P < 0.001). PMID- 7486936 TI - Early detection of rifampin in human nerve tissue after an oral dose of 600 milligrams. AB - Rifampin in picogram quantities inhibited the ability of Mycobacterium bovis 44 BCG P3 to release 14CO2 from the oxidation of [14C]palmitic acid. By using these mycobacteria in a bioassay, samples of serum and posterior tibial nerve were assayed for inhibitory concentrations of rifampin. Within 8 to 12 h after ingestion of 600 mg of rifampin, the drug was detected in eight patients in concentrations ranging from 0.52 to 4.1 micrograms/ml in serum and in concentrations ranging from 0.6 to 6.3 ng/mg in posterior tibial nerve fiber tissue. PMID- 7486938 TI - Pharmacokinetics of azithromycin in pediatric patients with acute otitis media. AB - The objective of our study was to characterize the pharmacokinetics of azithromycin after the oral administration of multiple doses in suspension to children with acute otitis media. Thirteen children (ranging in age from 7.5 months to 5 years) received a single oral dose of 10 mg of azithromycin per kg of body weight on day 1 followed by single daily doses of 5 mg/kg on days 2 to 5. Each child fasted overnight before receiving the final dose on day 5. Multiple blood samples were collected after the last dose. Concentrations of azithromycin in serum were measured by a specific high-performance liquid chromatography-mass spectrometry method. The means and standard deviations for the maximum concentration of azithromycin in serum, the time to maximum concentration of azithromycin in serum, the area under the concentration-time curve (from 0 to 24 h), and the elimination half-life were 224 +/- 120 ng/ml, 1.8 +/- 0.4 h, 1,841 +/ 651 ng.h/ml, and 31.6 +/- 6.6 h, respectively. Concentrations in serum (means +/ standard deviations) at 0 h (predose) and at 24, 48, and 72 h after the final dose were 51 +/- 26, 47 +/- 21, 27 +/- 17, and 17 +/- 13 ng/ml, respectively. Thus, the once-daily administration of azithromycin resulted in sustained systemic exposure to the drug. The drug dosage regimen used in this study should lead to tissue drug concentrations exceeding the MICs for common pathogens. PMID- 7486937 TI - Cloning and characterization of the Pseudomonas aeruginosa pbpB gene encoding penicillin-binding protein 3. AB - Clones containing the pbpB gene which encodes penicillin-binding protein (PBP) 3 of Pseudomonas aeruginosa were detected by hybridization by PCR amplification with primers based on the conserved sequences of high-molecular-weight PBPs. The translated amino acid sequence demonstrated 45% identity and had a total of 66% conserved amino acids relative to the Escherichia coli PBP3. The pbpB gene was located upstream of a gene homologous to the E. coli murE gene, which encodes uridine diphosphate-N-acetyl muramic acid-tripeptide synthetase. The overexpressed pbpB gene product reacted with 3H-penicillin G and had an apparent molecular weight of 60,000. PMID- 7486939 TI - Effects of Asp-179 mutations in TEMpUC19 beta-lactamase on susceptibility to beta lactams. AB - To examine the effect of disruption of the salt bridge (between Arg-164 and Asp 179 [numbering of Ambler et al. (Biochem J. 267:269-272, 1991)]) that anchors the conserved omega-loop in class A beta-lactamases, we obtained mutant enzymes with each of the 19 other amino acid residues replacing Asp-179 in the TEM beta lactamase encoded by pUC19 and studied the level of resistance to various beta lactams conferred by each enzyme. All mutations of Asp-179 compromised the level of resistance to ampicillin, but most of them enhanced resistance to ceftazidime. In contrast, mutations of Asp-179 generally impaired the low levels of resistance to cefepime and aztreonam. One might expect to find clinical isolates with mutant TEM beta-lactamases with replacements of Asp-179 that express an expanded spectrum of resistance to beta-lactams including ceftazidime. PMID- 7486940 TI - OXA-14, another extended-spectrum variant of OXA-10 (PSE-2) beta-lactamase from Pseudomonas aeruginosa. AB - Pseudomonas aeruginosa 455, isolated in Ankara, Turkey, produced a pI 6.2 beta lactamase determined by plasmid pMLH53 and resisted all beta-lactams except carbapenems. This beta-lactamase, named OXA-14, corresponded to OXA-10 (PSE-2) except that aspartate replaced glycine at position 157 and thus is intermediate between OXA-10 and OXA-11, which has aspartate at position 157 and a further substitution at position 143. PMID- 7486941 TI - Liposomal amphotericin B therapy of murine histoplasmosis. AB - Liposomal amphotericin B (AmBisome) was compared with amphotericin B deoxycholate for the treatment of disseminated murine histoplasmosis. Liposomal amphotericin B was well tolerated and, milligram for milligram, was as potent as amphotericin B deoxycholate. PMID- 7486943 TI - Influence of hemodialysis on plasma concentration-time profiles of mefloquine in two patients with end-stage renal disease: a prophylactic drug monitoring study. AB - Prophylactic drug monitoring of mefloquine and its carboxylic acid metabolite were studied in two patients with end-stage renal disease undergoing long-term hemodialysis treatment. The patients, short-term travellers to areas where malaria is endemic, took 250 mg of mefloquine (Lariam) once weekly for 2 weeks before and during their 3-week stay abroad and for one week after their return. Pre- and postdialysis blood samples were drawn before their departure and after their return. The concentration-time profiles of mefloquine and its metabolite in plasma samples taken before and after the 3- to 4-h dialysis sessions were similar. Mefloquine and its metabolite could not be detected in the dialysate. These findings show that mefloquine and its metabolite are not, or are very poorly, removed by hemodialysis. Concentrations in plasma and accumulation kinetics were similar to those reported for healthy volunteers and were associated with high prophylactic efficacy against malaria. No special dosage adjustments have to be made in patients undergoing hemodialysis treatment to achieve concentrations in plasma similar to those in healthy volunteers. The prophylactic dose of mefloquine could be given before, during, or after the hemodialysis session. PMID- 7486942 TI - K65R mutation of human immunodeficiency virus type 1 reverse transcriptase encodes cross-resistance to 9-(2-phosphonylmethoxyethyl)adenine. AB - Cloned variants of human immunodeficiency virus type 1 that contain the K65R mutation in reverse transcriptase have previously been shown to display approximately 10- to 30-fold resistance against 2',3'-dideoxycytidine, 2',3' dideoxyinosine, and 2',3'-dideoxy-3'-thiacytidine. On the basis of tissue culture studies with both primary T cells and established cell lines, we now report that the K65R mutation confers approximately 12- to 15-fold resistance to 9-(2 phosphonylmethoxyethyl)adenine (PMEA). Likewise, a chain termination system revealed that mutated recombinant K65R reverse transcriptase displays resistance to PMEA diphosphate, the active metabolite of PMEA, in cell-free enzyme assays. Parallel studies have shown that the M184V mutation in reverse transcriptase, associated with high-level resistance against the (-) enantiomer of 2',3'-dideoxy 3'-thiacytidine, does not confer resistance to PMEA in tissue culture. Viruses and enzymes that included both the K65R and M184V mutations were resistant to PMEA and PMEa diphosphate, respectively, but only to the extent conferred by the K65R mutation alone. PMID- 7486944 TI - Timing of penicillin treatment influences the course of Streptococcus pneumoniae induced middle ear inflammation. AB - Penicillin treatment timing, using the chinchilla pneumococcal otitis media model, was investigated. Early treatment (when approximately 10(3) pneumococcal CFU/ml was present in middle ear fluid) significantly accelerated inflammatory cell influx, whereas late treatment (with approximately 10(7) pneumococcal CFU/ml present in middle ear fluid) did not. Therefore, antiinflammatory interventions will be needed early in the course of inflammation. PMID- 7486946 TI - Effects of zinc-desferrioxamine on Plasmodium falciparum in culture. AB - The zinc-desferrioxamine (Zn-DFO) complex is considered to be more permeative into parasitized erythrocytes than is the metal-free DFO. The former may penetrate the cell and exchange its bound zinc for ferric ions, rendering the iron unavailable for vital parasite functions. The effects of these compounds on the in vitro development of Plasmodium falciparum are compared. The results indicate that Zn-DFO is superior to DFO, especially at concentrations below 20 microM, as shown by decreased levels of hypoxanthine incorporation, lower levels of parasitemia, and interference with the life cycle of the parasite. At low concentrations, DFO even enhanced parasite growth. Such an enhancement was not observed following exposure to Zn-DFO. Experiments in which the compounds were removed from the cultures indicated that parasites treated with Zn-DFO are less likely to recover at a later stage. Since DFO has already been used in humans for the treatment of malaria, its complex with zinc, which is more effective in vitro, should also be examined in vivo. PMID- 7486945 TI - Levels of flurithromycin in female genital tissue. AB - The levels of flurithromycin in gynecological tissue in 20 female patients were studied after preoperative administration. The tissue flurithromycin levels obtained were comparable to those obtained in serum at 3 and 4 h but were frequently higher than those in serum at 6 and 12 h. Flurithromycin reached the highest concentrations in ovary at 4 h and in endometrium at 6 h. PMID- 7486947 TI - Ex vivo effect of azithromycin in human leukocyte bactericidal functions. PMID- 7486948 TI - Minireview: nucleotide prodrugs. AB - Nucleotides have shown interesting biological activities in a wide variety of antiviral, antiproliferative, immunomodulatory and other biological assays, and they present promising drug candidates. Because of their negative charge(s) nucleotides suffer from some disadvantages which can be successfully overcome by the utilization of nucleotide prodrugs. Nucleotide prodrugs were successfully used to increase oral absorption of nucleotides in vivo. By taking advantage of intracellular triggers (reducing potential, enzyme activity, pH), nucleotide prodrugs can be used in vitro for the intracellular delivery of the nucleotide resulting in enhanced potency and in some cases enhanced selectivity. Nucleotide prodrugs have also been utilized for tissue specific delivery of the nucleotides in vivo resulting in altered selectivity and reduced toxicity. For nucleotide prodrugs, their ultimate intended use is (in most cases) in vivo for the treatment of a disease. Thus, it is important to incorporate adequate assays and design criteria into any prodrug effort. In vivo systems are complicated because of metabolism, excretion and tissue distribution of the prodrug and the parent. Thus, results of in vitro assays have to be interpreted cautiously because they may be unsuitable predictors of the in vivo situation. PMID- 7486953 TI - Analysis of mutations in the thymidine kinase gene of varicella zoster virus associated with resistance to 5-iodo-2'-deoxyuridine and 5-bromo-2'-deoxyuridine. AB - We have analyzed mutations in the thymidine kinase (TK) gene of varicella zoster virus (VZV) which showed resistance to 5-iodo-2'-deoxyuridine (IDU) and 5-bromo 2'-deoxyuridine (BrDU). Through sequencing of the TK gene, we found three amino acids were exchanged (41 Asn-->Ser, 266 Cys-->Ile, 288 Ser-->Leu). These mutations were not located at either the nucleoside- or the ATP-binding site. This result suggests that the resistance to IDU and BrDU in this particular strain is due to the change in conformation of TK rather than the replacement of amino acids in the binding sites. PMID- 7486949 TI - Suppression of infectious virus spread to the liver by foscarnet following lethal infection of acyclovir-resistant herpes simplex virus type 2 in mice. AB - Patients with the acquired immune deficiency syndrome (AIDS) occasionally develop hepatitis, pneumonia or esophagitis due to herpes simplex virus type 2 (HSV-2) infection. HSV hepatitis is a rare but serious complication in liver transplantation. Acyclovir-resistant HSV strains may emerge in immunocompromised patients. Following intraperitoneal inoculation, HSV-2 induces necrotizing hepatitis in mice. We studied the virus spread and mortality following intraperitoneal inoculation of HSV-2 RK (an acyclovir-resistant recombinant virus with altered thymidine kinase activity) as compared to its parent virus 8620K. Neither the 50% lethal dose (LD50) nor the average survival time was significantly different between the two strains. Parenteral acyclovir treatment was found to be effective against 8620K but not RK infection. Parenteral foscarnet treatment was effective against both RK and 8620K, and also inhibited the spread of either virus to the liver, spinal cord and brain. Peroral foscarnet administration was found to prevent the virus growth in the liver. PMID- 7486952 TI - A simple and rapid method for preliminary evaluation of in vivo efficacy of anti HIV compounds in mice. AB - In vivo efficacy of anti-HIV compounds is affected by various factors such as bioavailability, metabolism, clearance, and toxicity. Here we report a simple and rapid method that might be useful for preliminary evaluation of in vivo efficacy of anti-HIV compounds. MT-4 cells carrying proviral HTLV-1 were infected with HIV 1 in vitro and injected into the peritoneal cavity of SCID mice or BALB/c mice. Inoculated cells survive for 1-2 days, and support one to two cycles of viral replication which can be monitored by RT activity or p24 content in the supernatants of peritoneal wash fluids. Test compounds were administered either orally or subcutaneously. AZT, DDC and DDI, the nucleoside-type RT inhibitors currently in clinical use, all showed potent anti-HIV-1 activities in this mouse/MT-4 assay. HEPT (E-EBUdM), a non-nucleoside RT inhibitor, also showed potent anti-HIV-1 activity in vivo, whereas TIBO (R82913), another non-nucleoside RT inhibitor, was less active. In protease inhibitors KNI-272 and Ro 31-8959 showed good in vivo activities, while KNI-144, a compound closely related to KNI 272, showed poor in vivo activity. This mouse/MT-4 assay, although having a number of shortcomings as an animal model for HIV-1 infection, may be of some practical utility for preliminary evaluation of in vivo efficacy of potential anti-HIV compounds. PMID- 7486955 TI - 4-Guanidino-Neu5Ac2en fails to protect chickens from infection with highly pathogenic avian influenza virus. AB - The effectiveness of the novel sialidase inhibitor 4-guanidino-Neu5Ac2en, which is highly effective in mouse and ferret models of influenza virus infection (von Itzstein et al. (1993) Nature 363, 418-423), has been assessed as a prophylactic agent in the prevention of infection of chickens with highly pathogenic avian influenza viruses. At best a small delay in the onset of pyrexia and death was observed with one strain of fowl plague virus, but not with two other strains. These results demonstrate that a locally acting drug may be ineffective if virus can escape from the site of inoculation and replicate elsewhere. PMID- 7486950 TI - Effect of chloro-, cyano-, and amidino-substituted flavanoids on enterovirus infection in vitro. AB - Synthetic flavans, isoflavans and isoflavenes substituted with chloro, cyano and amidino groups were tested for their in vitro activity against poliovirus type 2, Coxsackie virus B4, echovirus type 6 and enterovirus 71. Plaque-reduction assays showed that substituted 3-(2H)-isoflavenes, carrying a double bond in the oxygenated ring, possess antiviral activity higher than that of the corresponding isoflavans. The most effective compounds were 4'-chloro-6-cyanoflavan and 6 chloro-4'-cyanoflavan. Studies on the mechanism of action of these two compounds suggested an effect on the early stages of viral replication. PMID- 7486951 TI - Efficacy of Coxiella burnetii and its chloroform-methanol residue (CMR) fraction against Rift Valley fever virus infection in mice. AB - Strains of Coxiella burnetii phase I and II whole cells (WC-I and WC-II) or whole cell fractions were assessed for their potential to induce long-lasting protection in endotoxin-non-responder C3H/HeJ or CD-1 mice against Rift Valley fever (RVF) virus challenge. Among the whole cell fractions, only the chloroform methanol residue (CMR), administered as a single dose (100 micrograms per mouse) 24 h before viral challenge, effectively protected 100% of the mice from RVF virus; the CMR of the Ohio strain of C. burnetii was not protective. Most of the RVF virus-infected mice treated with other C. burnetii cell fractions died, although their times to death varied. Lipopolysaccharide (LPS) associated with CMR preparations used in these studies, did not protect against RVF virus challenge. A single dose of 100 micrograms of CMR given 24 h before viral challenge completely eradicated 4-5 logs of RVF virus in the serum, liver, spleen, and central nervous system. Compared to several other immunomodulators, CMR was an equally effective antiviral agent. Efficacy of CMR of both Henzerling and Ohio strains disappeared or was marginal when treatment was initiated 2-3 days before RVF viral challenge, even when a second or a third dose of CMR was administered after challenge. A single dose of liposome-encapsulated CMR to RVF virus-infected mice extended the range of therapeutic efficacy of this biologically active component of C. burnetii to 4 days before infection. PMID- 7486954 TI - Reversion of duck hepatitis B virus DNA replication in vivo following cessation of treatment with the nucleoside analogue ganciclovir. AB - In order to define, in more detail, the virological events which occur after completion of antiviral chemotherapy, ducks congenitally infected with the duck hepatitis B virus (DHBV) were treated for 4 weeks with the nucleoside analogue ganciclovir and followed up over a 7-day period. Specimens of serum and liver were collected daily during follow-up for virological analysis. Treatment resulted in a substantial reduction in both viraemia and liver DHBV DNA replicative intermediates. However, after cessation of treatment, viraemia returned to detectable levels within 4 days. In the liver, the viral supercoiled DNA (SC DNA) was the form least affected by therapy and returned to near control levels by day 2 post-treatment. The other hepatic replicative intermediates reached pretreatment levels within 4 days of cessation of therapy. This study has defined the kinetics of relapse of viral replication after completion of antiviral therapy in the duck hepatitis B model. Of all viral replicative forms, the SC DNA appears to be the one which is most resistant to nucleoside analogue therapy and is presumably responsible for the relapse phenomenon observed post treatment. PMID- 7486957 TI - Pharmacokinetics and toxicity of the human immunodeficiency virus inhibitor 1 ethoxymethyl-6-phenylselenenyl-5-ethyluracil in rodents. AB - 1-(Ethoxymethyl)-6-(phenylselenenyl)-5-ethyluracil (E-EPSeU) has been shown to exhibit potent and selective activity against human immunodeficiency virus type 1 in vitro. The pharmacokinetics of E-EPSeU were characterized after intravenous administration of 5, 10 and 15 mg/kg to rats. Plasma and urine concentrations of E-EPSeU were determined by HPLC. The plasma protein binding of E-EPSeU averaged 86 +/- 4% and the blood: plasma concentration ratio was unity. E-EPSeU concentrations after the 5 mg/kg dose were too low to reliably characterize the pharmacokinetics. The pharmacokinetics of E-EPSeU were independent of dose over the range of 10-15 mg/kg. Plasma concentrations of E-EPSeU declined in a bi exponential manner with terminal half-life of 0.45 +/- 0.12 h (mean +/- S.D.). The steady-state volume of distribution was 0.091 +/- 0.031 1/kg, suggesting the compound distributed primarily into blood. The systemic clearance (0.63 +/- 0.13 1/h/kg) was moderate and limited, in part, by protein binding. No parent compound was detected in urine. E-EPSeU-related toxicities were observed at high doses. One rat, out of 5, died 4 h after 15 mg/kg of E-EPSeU was administered and two rats administered 20 and 25 mg/kg died within 1 h. Two mice, out of 5, administered 30 mg/kg/day of E-EPSeU intraperitoneally for 6 days died during the experiment, while significant loss of body weight was observed in the surviving mice. However, body weight of the surviving mice returned to control values within 2 weeks after E-EPSeU treatment was stopped.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7486956 TI - Efficacy of traditional herbal medicines in combination with acyclovir against herpes simplex virus type 1 infection in vitro and in vivo. AB - Traditional herbal medicines have been safely used for the treatment of various human diseases since ancient China. We selected 10 herbal extracts with therapeutic antiherpes simplex virus type 1 (HSV-1) activity. Among these, Geum japonicum Thunb., Rhus javanica L., Syzygium aromaticum (L.) Merr. et Perry, or Terminalia chebula Retzus showed a stronger anti-HSV-1 activity in combination with acyclovir than the other herbal extracts in vitro. When acyclovir and/or a herbal extract were orally administered at doses corresponding to human use, each of the 4 combinations significantly limited the development of skin lesions and/or prolonged the mean survival times of infected mice compared with both acyclovir and the herbal extract alone (P < 0.01 or 0.05). These combinations were not toxic to mice. They reduced virus yields in the brain and skin more strongly than acyclovir alone and exhibited stronger anti-HSV-1 activity in the brain than in the skin, in contrast to acyclovir treatment by itself. Combinations of acyclovir with historically used herbal medicines showed strong combined therapeutic anti-HSV-1 activity in mice, especially reduction of virus yield in the brain. PMID- 7486958 TI - Inhibition of visna virus replication and cytopathic effect in sheep choroid plexus cell cultures by selected anti-HIV agents. AB - Several anti-HIV agents were tested against visna virus replication and cytopathic effect (CPE) in sheep choroid plexus cell cultures. Sulphated polysaccharides (i.e., dextran sulphate, pentosan polysulphate and heparin) and plant lectins, which inhibit viral adsorption and fusion, were found to be 10- to 40-fold less active against visna virus than against HIV. Bicyclam derivatives were at least 250-fold less active against visna virus and the highly HIV-1 specific TIBO derivatives were without a significant inhibitory effect on visna virus at subtoxic concentrations. In contrast, several 2',3'-dideoxynucleosides and acyclic nucleoside phosphonate analogues, which inhibit reverse transcription, were found to be very effective inhibitors of visna virus replication and viral CPE in cell culture. PMID- 7486959 TI - Evaluation of the antiviral activity of N-(phosphonoacetyl)-L-aspartate against paramyxoviruses in tissue culture and against respiratory syncytial virus in cotton rats. AB - N-(phosphonoacetyl)-L-aspartate (PALA), a potent inhibitor of L-aspartic acid transcarbamoylase, was evaluated for cytotoxicity and antiviral activity against three different paramyxoviruses in tissue culture, and for antiviral efficacy and toxicity in vivo using a cotton rat-respiratory syncytial virus (RSV) model. Significant in vitro cytotoxicity was observed in proliferating cultures of HEp-2 (IC50 = 250 micrograms/ml) and Vero cells (IC50 = 32 micrograms/ml), but was less evident in cultures containing confluent monolayers (i.e., stationary cells) of these cells, or in cultures of Madin Darby canine kidney (MDCK) cells (these IC50 values were all > or = 750 micrograms/ml, with 1000 micrograms/ml being the maximum concentration tested). Mean selective indices (ratio of the median cytotoxic dose: median efficacious dose) of 1, 72 and 146 were obtained against parainfluenza virus type 3, RSV and measles virus, respectively, when PALA was tested against these viruses using confluent HEp-2 and Vero cell monolayers. In cotton rats, significant reductions in pulmonary titers (0.8-1.4 log10/g lung) compared to pulmonary viral titers in placebo-treated control animals, were consistently seen in cotton rats given > or = 10 mg of PALA/kg/day (b.i.d.) intraperitoneally on days 1-3 postinfection with either subtype A or B RSV. No toxic effects were noted even in animals given 100 mg of PALA/kg/day for 7 consecutive days. PMID- 7486960 TI - Antiviral activity of phosphonoformate on rotavirus transcription and replication. AB - The antiviral effect of foscarnet (PFA) on the replication of rotavirus, a member of the Reoviridae, was studied. The pyrophosphate analogue is an effective inhibitor of several viral polymerases acting on the enzyme pyrophosphate binding site. Replication of rotavirus in MA104 cells using different u.o.i. was inhibited by PFA in a concentration dependent manner, due to the inhibition of both plus- and minus-strand RNA synthesis. The addition of PFA to infected cells was specific for the inhibition of viral replication since uninfected cell incubated at the same PFA concentrations did not exhibit any cytotoxic effect. The 50% inhibitory effect of PFA on in vitro mRNA synthesis was obtained at a concentration of 150 microM. Over 80% of the in vitro minus-strand RNA synthesis was inhibited at a concentration of 320 microM, when PFA was assayed using replicase-enriched cell infected fraction. The results suggest that the effect may be due to an interaction of PFA with the viral polymerase, since this protein catalyses both plus- and minus-strand RNA synthesis. The results of experiments using the temperature-sensitive viral polymerase mutant show that the mutant is less sensitive to PFA, suggesting that this polypeptide is the target for PFA. PMID- 7486961 TI - Efficacy of AZT therapy in reducing p24 antigen burden in a modified SCID mouse model of HIV infection. AB - A modified severe combined immunodeficient (SCID) mouse model of HIV infection which utilized multiple reconstitutions with human lymphocytes and a large inoculum of HIV was investigated. This mouse model yielded splenic HIV p24 antigen concentrations detectable by standard clinical means. The concentration of p24 exceeded 600 pg/g of spleen through 4 weeks postinfection. A 1-week course of AZT therapy initiated after infection produced a dose responsive reduction in p24 antigen burden. Up to a 95% reduction in p24 antigen burden was observed following AZT therapy at 50 mg/kg/day, while AZT therapy at 5 and 0.5 mg/kg/day produced 52 and 18% reductions. In vitro and pharmacokinetic evaluations correlated potency and tissue concentrations of AZT with treatment efficacy. Active HIV replication in the SCID mice was suggested by both the recovery of viable virus from SCID spleens, and by the efficacy of a brief course of AZT therapy. This SCID mouse model of HIV infection was more quantitative than previous mouse models that utilize PCR-based techniques for detection of HIV. The high HIV burden in this SCID mouse model allowed reductions in p24 concentration to be monitored in response to AZT therapy. A dose response to AZT therapy was demonstrated, even when the first dose was administered after infection. This result suggests greater sensitivity than in previous models in which pretreatment with AZT was required to produce a protective response. This SCID mouse model may be useful for determining efficacy of experimental HIV therapeutics prior to clinical use. An effective animal model could result in a reduction in cost and more rapid development of effective HIV therapeutics. PMID- 7486963 TI - Progress on AORN's strategic plan demonstrates our continued commitment. PMID- 7486966 TI - Surgical masks. PMID- 7486964 TI - Duplicate publication wastes readers' time, clutters the literature, violates copyright law, and delays publication of timely articles. PMID- 7486967 TI - Surgeons as clients. PMID- 7486962 TI - Active compounds from Saussurea lappa Clarks that suppress hepatitis B virus surface antigen gene expression in human hepatoma cells. AB - We have examined the antiviral activity of the crude extract prepared from the root of Saussurea lappa Clarks, a Chinese medicinal herb which is widely used for many illnesses including cancer. Two active components, costunolide and dehydrocostus lactone, were identified which show strong suppressive effect on the expression of the hepatitis B surface antigen (HBsAg) in human hepatoma Hep3B cells, but have little effect on the viability of the cells. Both costunolide and dehydrocostus lactone suppress the HBsAg production by Hep3B cells in a dose dependent manner with IC50s of 1.0 and 2.0 microM, respectively. Northern blotting analysis shows that the suppression of HBsAg gene expression by both costunolide and dehydrocostus lactone were mainly at the mRNA level. Furthermore, the suppressive effect of costunolide and dehydrocostus lactone on HBsAg and hepatitis B e antigen (HBeAg), a marker for hepatitis B viral genome replication in human liver cells, was also observed in another human hepatoma cell line HepA2 which was derived from HepG2 cells by transfecting a tandemly repeat hepatitis B virus (HBV) DNA. Similarly, the mRNA of HBsAg in HepA2 cells was also suppressed by these two compounds. Our findings suggest that costunolide and dehydrocostus lactone may have potential to develop as specific anti-HBV drugs in the future. PMID- 7486968 TI - Scalp replantation after traumatic injury. AB - A 30-year-old female patient lost her scalp in an industrial accident. Surgeons used a microscope to reconnect the arteries and veins in her scalp and to reattach the scalp to the patient's cranium. The patient tolerated the procedure without complications, although she developed venous congestion in her right ear while she was in the neuroscience intensive care unit. After several days of leech therapy, the patient experienced an uneventful postoperative course. One month after discharge from the hospital, the patient's hair was growing back in and her incision lines were fading. PMID- 7486965 TI - RN first assistant program. PMID- 7486970 TI - Pediatric fibromatosis requiring mandibular resection and reconstruction. AB - Fibromatosis is the nonmalignant proliferation of fibroblasts that aggressively invade adjacent tissues. The cause of this abundant growth is unknown. When fibromatosis develops in the mandible or adjoining tissues, its aggressive growth can compress the trachea and cause death. Resection is the treatment of choice. Mandibular reconstruction is performed to prevent displacement of the remaining mandibular segment or segments and consequent difficulties in eating and speaking. Perioperative nurses who care for children with oral or paraoral fibromatoses must understand the special physical and psychological needs of these patients to effectively plan their care. PMID- 7486969 TI - Dental restorations using titanium osseointegrated dental implants. AB - Dental implantology (ie, the science of replacing teeth) has evolved from the use of crude, unstable, natural materials to the use of titanium osseointegrated dental implants and sophisticated prosthetic appliances. Dental prosthetics are used in an increasing number of routine dental restorations. Postoperative patients who have undergone dental restorations using titanium osseointegrated dental implants enjoy results that approximate the look and feel of natural teeth. Optimal patient outcomes are best accomplished through a multidisciplinary team approach, and perioperative goals of patient commitment and compliance are achieved through comprehensive education. PMID- 7486971 TI - Dental surgery standards for perioperative nurses. Medical Center of Central Georgia. AB - Practice standards for dental surgery are necessary to maintain quality care for dental patients. Specific standards or recommended practices for dental surgery have not been addressed by AORN, and none are available from the American Dental Association or the Georgia Dental Association. We incorporated the Centers for Disease Control and Prevention guidelines, and Occupational Safety and Health Administration regulations had to be incorporated into existing perioperative standards to institute dental surgery standards for our facility. The standards for dental surgery at the Medical Center of Central Georgia, Macon, evolved with the leadership of perioperative nurses. PMID- 7486972 TI - Microsurgical transfer of vascularized tissue to close problem wounds. AB - Interventions to achieve optimal wound healing challenge the skills of all health care professionals. Although most wounds are best managed by appropriate nursing intervention outside the OR, some complex wounds require more expeditious closure to ensure maximum preservation of vital structures and their functions. Microsurgical transfer of vascularized tissue (ie, flaps) often is the only method of achieving immediate wound coverage. Perioperative team members who treat such wounds must be aware of the distinct attributes and potential morbidity associated with flap procedures. PMID- 7486973 TI - Evaluating a revised self-efficacy scale for preoperative patients. AB - Nurses conduct preoperative teaching to help patients cope with the adversities of surgery. Self-efficacy can predict an individual's behavior in aversive situations (eg, surgery); therefore, assessing patients' self-efficacy is one way perioperative nurses can plan patient care and help patients through the surgical experience. This study expands on the results of a previous evaluation of a preoperative self-efficacy scale. The content and face validity of the revised 15 item scale used in this study were assessed before the instrument was administered to preoperative patients. Evidence supports the validity of the multidimensional instrument. Internal consistency estimates > or = .84 support the subscales' reliability. PMID- 7486974 TI - A surgical services learning needs assessment. AB - Staff development educators are challenged to meet the learning needs of a variety of subspecialties within surgical services departments. A formal learning needs assessment survey, based on the individual self-fulfillment model, provides a means to identify the educational needs of staff members. Using questionnaires constructed for various types of staff members within a surgical services department, the author assessed staff members' clinical, interpersonal, and sociopolitical learning needs. The response rate was 56%, and the identified needs were legal issues, research, equipment (eg, ultrasonic aspirators, pressure monitors), and procedures (eg, cerebral aneurysm repairs, arteriovenous malformation corrections). Limitations included self-identification of needs and a low response rate from one subspecialty; however, the author concludes that the survey format provides an effective means of identifying staff members' learning needs. PMID- 7486975 TI - Does perioperative nursing include caring? AB - Does perioperative nursing include caring? Does it show evidence of a theoretical base of caring? Yes, most definitely. We touch; we listen; we hold; we act as advocates. The challenge is to articulate these actions in our body of knowledge and practice. What would an outcome statement on perioperative caring look like? As perioperative critical paths are developed, how will perioperative caring outcomes be expressed? These are the challenges perioperative nurses must meet to demonstrate the necessity of the perioperative nursing role in providing humanistic care in the OR. PMID- 7486977 TI - Proposed recommended practices for traffic patterns in the perioperative practice setting. Association of Operating Room Nurses. PMID- 7486976 TI - A successful device tracking program. PMID- 7486978 TI - Nurse Education Act and National Institute for Nursing Research funding in jeopardy. PMID- 7486980 TI - A simple method for quantifying activity and survival of microorganisms involved in bioremediation processes. AB - We have developed a substrate-induced growth response (SIGR) method for quantifying activity and population dynamics of microorganisms involved in bioremediation processes in soil and bioreactors. The biomass of organisms that can mineralize a given chemical can be estimated based on the concentration of that chemical needed to induce the growth of the standing population. Estimates of population size are obtained by using nonlinear regression techniques to fit a simple model of microbial population dynamics to biodegradation curves. Using this approach we obtain estimates of values for parameters such as initial population size and growth rate of organisms carrying out biodegradative processes. Our approach was validated by comparing model parameter estimates with independent estimates of the same parameters from the same bioremediation systems. Examples studied include pentachlorophenol degraders introduced into soil and 2,4-dinitrophenol degrading organisms in a bioreactor. PMID- 7486979 TI - Large-scale oligonucleotide synthesizers. I. Basic principles and system design. AB - The central problem in scaling up oligonucleotide synthesis is to expose each element of a large bed to the same conditions obtaining in very small ones, for the same intervals of time. Our analysis suggests that scale-up is chiefly limited by fluid path length through the bed. By using annular beds in zonal centrifuge rotors of unique design, with fluid flow controlled by combining centrifugal force with differences in physical density between reagents, reagent fronts may be kept exactly perpendicular to the direction of flow in each bed element. Under these conditions, bed volume may be increased by increasing rotor length and diameter. The rotor is lined with polypropylene or Teflon, and has a thick tempered glass end window. Transparent rotary valves of a unique design allow any of 47 reagents to be selected and the direction of flow through the rotor to be controlled. A photodiode spectrophotometer provides complete absorption spectra on fluid in the rotor inlet and outlet lines every 4 s, and a large balance weighs effluent from the synthesizer continuously. The entire operation is controlled by a work station, and steps may be programmed by time, absorbance, or reagent mass. Reagents are identified by spectra, and trityls are integrated on line. A detailed time-stamped log file provides a complete record of each synthesis. PMID- 7486981 TI - Use of a pentachlorophenol degrading bacterium to bioremediate highly contaminated soil. AB - A Sphingomonas species that mineralizes high concentrations of pentachlorophenol (PCP) was isolated from a PCP-contaminated EPA Superfund site. This bacterium, identified as Sphingomonas sp. strain RA2, is able to degrade PCP at concentrations of up to 300 micrograms/mL in liquid culture. This organism was tested for its ability to degrade high concentrations of PCP in a soil that did not contain organisms capable of degrading high concentrations of PCP. When inoculated into contaminated soil, Sphingomonas sp. RA2 mineralized PCP at concentrations of 300, 600, 900, and 1200 micrograms PCP/g of soil, but was unable to mineralize 1500 micrograms PCP/g of soil. Only very minimal loss of PCP was seen in uninoculated soils. The results of this study demonstrate that Sphingomonas sp. RA2 may be a useful organism for remediation of sites contaminated with high concentrations of PCP. PMID- 7486982 TI - Molecular site assessment and process monitoring in bioremediation and natural attenuation. off. AB - A variety of modern biotechnical approaches are available to assist in optimizing and controlling bioremediation processes. These approaches are broad-ranging, and may include genetic engineering to improve biodegradative performance, maintenance of the environment, and process monitoring and control. In addition to direct genetic engineering strategies, molecular diagnostic and monitoring technology using DNA gene probing methods and new quantitative mRNA analytical procedures allows direct analysis of degradative capacity, activity, and response under in situ conditions. Applications of these molecular approaches in process developments for trichloroethylene (TCE), polychlorinated biphenyls (PCB), and polynuclear aromatic hydrocarbons (PAH) bio-oxidation in soils, aquifer sediments, and ground-water treatment reactors have been demonstrated. Molecular genetic technologies permit not only the development of new processes for bioremediation, but also new process monitoring, control strategies, and molecular optimization paradigms that take full advantage of vast and diverse abilities of microorganisms to destroy problem chemicals. PMID- 7486983 TI - Environmental aspects of PAH biodegradation. AB - Polycyclic aromatic hydrocarbons (PAHs) are ubiquitous pollutants, some of which are on the US Environmental Protection Agency priority pollutant list. Consequently, timely clean-up of contaminated sites is important. The lower-mol wt PAHs are amenable to bioremediation; however, higher-mol-wt PAHs seem to be recalcitrant to microbial degradation. The rates of biodegradation of PAHs are highly variable and are dependent not only on PAH structure, but also on the physicochemical parameters of the site as well as the number and types of microorganisms present. PAHs sorb to organic matter in soils and sediments, and the rate of their desorption strongly influences the rate at which microorganisms can degrade the pollutants. Much of the current PAH research focuses on techniques to enhance the bioavailability and, therefore, the degradation rates of PAHs at polluted sites. Degradation products of PAHs are, however, not necessarily less toxic than the parent compounds. Therefore, toxicity assays need to be incorporated into the procedures used to monitor the effectiveness of PAH bioremediation. In addition, this article highlights areas of PAH research that require further investigation. PMID- 7486984 TI - Ribozyme mimics as catalytic antisense reagents. AB - Viral and fungal infections and some cancers may be described as diseases that are characterized by the expression of certain unwanted proteins. They could be termed induced genetic disorders, with induction provided by mutation or infection. A comprehensive method to inactivate injurious genes based on their nucleic acid sequences has the potential to provide effective antiviral and anticancer agents with greatly reduced side effects. We describe a chemical approach to such gene-specific pharmaceutical agents. Our initial efforts have been to develop new chemical reagents that can carry out catalytic destruction of specific mRNA sequences. We chose hydrolysis as a chemical means of destruction, because hydrolysis is compatible with living cells. Our sequence-specific catalytic RNA hydrolysis reagents may be described as functional ribozyme mimics. Reactivity is provided by small-molecule catalysts, such as metal complexes. Specificity is provided by oligonucleotide probes. Here we report initial results on the sequence-specific, hydrolytic cleavage of mRNA from the HIV gag gene, using a ribozyme mimic. The reagent is composed of a terpyridylCu(II) complex for cleavage activity and an oligonucleotide for sequence specificity. PMID- 7486985 TI - Colocalizing ribozymes with substrate RNAs to increase their efficacy as gene inhibitors. AB - The ability to target ribozymes to specifically cleave viral RNAs in vitro has led to much speculation about their potential therapeutic value as antiviral agents in vivo. To transfer a ribozyme's potential as an antiviral agent from test tubes to cells and organisms successfully, the characteristics that distinguish these settings must be considered. In vitro, ribozymes and substrate RNAs freely diffuse in solution in test tubes, and trans-cleavage reactions are dependent on a diffusive step. In eukaryotic cells, by contrast, many RNAs do not appear to diffuse freely. Instead, they appear to be highly compartmentalized and actively sorted to specific cellular locations. Such RNA trafficking may result in localization of substrate RNAs in a different compartment than ribozymes, which would effectively reduce substrate RNA availability to ribozymes and therefore limit the effectiveness of ribozymes as gene inhibitors. PMID- 7486986 TI - A stereotypic, transplantable liver tissue-culture system. AB - A method of coculturing adult rat hepatic parenchymal cells (PC) and stromal cells in a three-dimensional framework of nylon filtration screens or biodegradable polymer meshes was developed in our laboratory. Rat liver stroma, which includes vascular and bile duct endothelial cells, fat-storing cells, fibroblasts, and Kupffer cells, were isolated by gradient centrifugation after in situ liver perfusion and expanded in monolayer culture prior to seeding onto nylon screens or bioresorbable polyglycolic acid (PGA) polymers oriented into a felt-like construct. A second inoculum of freshly isolated PC was applied after the stromal cells became established. Histological analyses revealed that PC proliferation occurred until all available space for expansion within the template was exhausted. These cells retained their rounded morphology, and after 4-5 wk 7-9 "layers" of PC filled the 140-microns deep template. Dioxin-inducible cytochrome P450 activity was detected for up to 58 d in culture, and albumin, fibrinogen, transferrin, and soluble fibronectin were detected in the medium by enzyme-linked immunosorbent assay (ELISA) for 48 d in vitro. Immunohistochemical analysis of sections through the cultures confirmed the presence of these proteins as well as cytokeratin at the cellular level; the extracellular matrix stained for both collagen type III and laminin. Long-term PC proliferation and function were enhanced by the presence of stromal cells as well as by a meshwork template whose geometry allows the interaction of PC with stroma and matrix on several different planes. To permit transplantation, cocultures of hepatic PC and stromal cells were established on PGA felt constructs instead of nylon screens. After approximately 24 d in vitro, these constructs were grafted into sites in the mesentery, omentum, and subcutaneous tissues of adult Long-Evans rats. The growth of hepatocytes after 30 d in situ was evident by histological analysis; grafts of cocultures regenerated a liver-like architecture consisting of sinusoids and putative biliary structures. In addition, PC at these extrahepatic graft sites were positive for albumin, transferrin, and fibrinogen synthesis by immunohistochemistry. Graft survival was enhanced when recipients were subjected to approximately 40% hepatectomy. Hepatic PC:stromal cell cocultures may prove useful in the restoration of liver function either by direct transplantation using PGA or similar templates, or as extracorporeal devices, using nylon screens. PMID- 7486987 TI - Production of human insulin in an E. coli system with Met-Lys-human proinsulin as the expressed precursor. AB - The construction of a gene encoding Lys-human proinsulin, its direct expression in E. coli, and the simple purification procedure are described here. The temperature inducible promotor was employed for induction in a very short time. The expression level could reach 20-30%. After simple downstream processing and only one step of Sephadex G50 purification, 150 mg recombinant Lys-human proinsulin with a purity of up to 90% could be obtained easily from 1 L of high density fermentation medium. The obtained product is in the form of Met-Lys-human proinsulin because of the failure of the bacterial host to remove the initiator methionine residue. The Lys-human proinsulin could be changed into human insulin by trypsin and carboxypeptidase B treatment in later steps. After separation with DEAE-Sephadex A25, human insulin with expected amino acid composition and full native biological activity could be obtained with a yield of 50 mg/L of fermentation medium. PMID- 7486988 TI - Genetic diversity and geographical distribution of wild Saccharomyces cerevisiae strains from the wine-producing area of Charentes, France. AB - Electrophoretic karyotyping, mitochondrial DNA restriction fragment length polymorphism analysis, and PCR amplification of interspersed repeats were used to study the variability, phylogenetic affinities, and biogeographic distribution of wild Saccharomyces cerevisiae enological yeasts. The survey concentrated on 42 individual wine cellars in the Charentes area (Cognac region, France). A limited number (35) of predominant S. cerevisiae strains responsible for the fermentation process have been identified by the above molecular methods of differentiation. One strain (ACI) was found to be distributed over the entire area surveyed. There seemed to be little correlation between geographic location and genetic affinity. PMID- 7486989 TI - Heat-tolerant methanotrophic bacteria from the hot water effluent of a natural gas field. AB - Methanotrophic bacteria were isolated from a natural environment potentially favorable to heat-tolerant methanotrophs. An improved colony plate assay was developed and used to identify putative methanotrophic colonies with high confidence. Fourteen new isolates were purified and partially characterized. These new isolates exhibit a DNA sequence homology of up to 97% with the conserved regions in the mmoX and mmoC genes of the soluble methane monooxygenase (MMO)-coding gene cluster of Methylococcus capsulatus Bath. The copper regulation of soluble MMO expression in the same isolates, however, differs from that of M. capsulatus Bath, as the new isolates can tolerate up to 0.8 microM copper without loss of MMO activity while a drastic reduction of MMO activity occurs already at 0.1 microM copper in M. capsulatus Bath. The isolates can be cultivated and utilized at elevated temperatures, and their copper- and heat-tolerant MMO activity makes these bacteria ideal candidates for future biotechnological use. PMID- 7486990 TI - Growth of the fungus Cladosporium sphaerospermum with toluene as the sole carbon and energy source. AB - The fungus Cladosporium sphaerospermum was isolated from a biofilter used for the removal of toluene from waste gases. This is the first report describing growth of a eukaryotic organism with toluene as the sole source of carbon and energy. The oxygen consumption rates, as well as the measured enzyme activities, of toluene-grown C. sphaerospermum indicate that toluene is degraded by an initial attack on the methyl group. PMID- 7486991 TI - Unidirectional motility of Escherichia coli in restrictive capillaries. AB - In a 6-microns capillary filled with buffer and in the absence of any chemotactic stimuli, Escherichia coli K-12 cells swim persistently in only one direction. This behavior of E. coli can be simply explained by means of the length and relative rigidity of their flagella. Single-cell motility parameters--swimming speed, turn angle, and run length time--were measured. Compared with the motility parameters measured in bulk phase, turn angle was influenced because of the effect of the geometrical restriction. PMID- 7486993 TI - Purification and characterization of an extracellular pectate lyase from an Amycolata sp. AB - The extracellular pectate lyase (EC 4.2.2.2) of a nonsporulating Amycolata sp. was purified to homogeneity by anion- and cation-exchange chromatographies followed by hydrophobic interaction chromatography. The enzyme cleaved polygalacturonate but not highly esterified pectin in a random endolytic transeliminative mechanism that led to the formation of a wide range of 4,5 unsaturated oligogalacturonates. As shown by high-performance anion-exchange chromatography and pulsed amperometric detection, these unsaturated oligogalacturonates were further depolymerized by the enzyme to the unsaturated dimer and trimer as final products. The pectate lyase had a molecular weight of 31,000 determined by sodium dodecyl sulfate-polyacrylamide gel electrophoresis and a molecular mass of 30,000 Da determined by matrix-assisted laser desorption ionization mass spectrometry. The isoelectric point of the protein was 10. Maximum activity occurred at pH 10.25. Calcium was essential for activity, and EDTA inactivated the enzyme under standard assay conditions. Interestingly, EDTA did not inhibit the ability of the enzyme to cleave the native pectin (protopectin) of ramie (Boehmeria nivea) fibers. The Km value with sodium polygalacturonate as the substrate was 0.019 g liter-1. The purified enzyme lost its activity after a 1-h incubation at 50 degrees C but was stabilized by calcium or polygalacturonate. The N-terminal sequence showed high similarity within a stretch of 13 amino acids to the N-terminal sequences of pectate lyases PLa and PLe from Erwinia chrysanthemi. The Amycolata sp. did not produce additional isozymes of pectate lyase but produced further activities of pectinesterase, xylanase, and carboxymethyl cellulase when grown in a medium with decorticated bast fibers from ramie as the sole carbon source. PMID- 7486992 TI - Molecular characterization of genes involved in the production of the bacteriocin leucocin A from Leuconostoc gelidum. AB - Leucocin A is a small heat-stable bacteriocin produced by Leuconostoc gelidum UAL187. A 2.9-kb fragment of plasmid DNA that contains the leucocin structural gene and a second open reading frame (ORF) in an operon was previously cloned (J. W. Hastings, M. Sailer, K. Johnson, K. L. Roy, J. C. Vederas, and M. E. Stiles, J. Bacteriol. 173:7491-7500, 1991). When a 1-kb DraI-HpaI fragment containing this operon was introduced into a bacteriocin-negative variant (UAL187-13), immunity but no leucocin production was detected. Leucocin production was observed when an 8-kb SacI-HindIII fragment of the leucocin plasmid was introduced into L. gelidum UAL187-13 and Lactococcus lactis IL1403. Nucleotide sequence analysis of this 8-kb fragment revealed the presence of three ORFs in an operon upstream of and on the strand opposite from the leucocin structural gene. The first ORF (lcaE) encodes a putative protein of 149 amino acids with no apparent function in leucocin A production. The second ORF (lcaC) contains 717 codons that encode a protein homologous to members of the HlyB family of ATP binding cassette transporters. The third ORF (lcaD) contains 457 codons that encode a protein with marked similarity to LcnD, a protein essential for the expression of the lactococcal bacteriocin lactococcin A. Deletion mutations in lcaC and lcaD resulted in loss of leucocin production, indicating that LcaC and LcaD are involved in production and translocation of leucocin A. The secretion apparatus for lactococcin A did not complement mutations in the lcaCD genes to express leucocin A in L. lactis.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7486994 TI - Recombinant infectious hematopoietic necrosis virus and viral hemorrhagic septicemia virus glycoprotein epitopes expressed in Aeromonas salmonicida induce protective immunity in rainbow trout (Oncorhynchus mykiss). AB - Fragments of the glycoprotein genes of viral hemorrhagic septicemia virus (VHSV) and infectious hematopoietic necrosis virus (IHNV) were cloned into a bacterial broad-host-range expression vector under the control of the plac promoter. Western blot (immunoblot) analysis with monoclonal antibodies specific to the glycoproteins demonstrated the inducible expression of the fusion proteins in Escherichia coli. Aeromonas salmonicida is the causative agent of furunculosis in salmonid fish. It was confirmed that an avirulent strain of A. salmonicida, A440, which contains a deletion in the structural gene for the paracrystalline surface protein array, will provide protective immunity against furunculosis when used as a live attenuated vaccine. The plasmid-encoded viral epitopes were then mobilized into A440 for use as a shuttle system for the expression of fragments of the glycoprotein genes of IHNV and VHSV. Vaccination of rainbow trout with A440 containing the viral epitopes resulted in the development of protective immunity against both VHSV and IHNV. This indicates that the use of cloned fragments of the glycoproteins and the use of A. salmonicida as a shuttle system constitute a feasible approach to fish vaccine development. PMID- 7486995 TI - Trehalose and sucrose protect both membranes and proteins in intact bacteria during drying. AB - The microorganisms Escherichia coli DH5 alpha and Bacillus thuringiensis HD-1 show an increased tolerance to freeze-drying when dried in the presence of the disaccharides trehalose and sucrose. When the bacteria were dried with 100 mM trehalose, 70% of the E. coli and 57% of the B. thuringiensis organisms survived, compared with 56 and 44%, respectively, when they were dried with sucrose. Only 8% of the E. coli and 14% of the B. thuringiensis organisms survived drying without the sugars. Fourier transform infrared spectroscopy was used to investigate the role of membrane phase transitions in the survival of the organisms during drying and rehydration. Both E. coli and B. thuringiensis showed an increase of 30 to 40 degrees C in the temperature of their phospholipid phase transition when dried without the sugars, while phase transition temperatures of those dried with the sugars remained near those of the hydrated cells. A Fourier transform infrared spectroscopy microscope made it possible to investigate the effects of drying on the protein structure in the intact cells. The amide II peak shifts from 1,543 cm-1 in the hydrated cells to about 1,533 cm-1 in the cells dried without sugar. There is no shift in the amide II peak when the cells are dried with trehalose or sucrose. We attribute the increased survival to the sugars' ability to lower the membrane phase transition temperature and to protect protein structure in the dry state.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7486997 TI - Mechanisms for the development of genetically variable mycorrhizal mycelia in the ectomycorrhizal fungus Laccaria bicolor. AB - An in vitro study investigated mechanisms for the development of genetically variable mycorrhizal mycelia for Laccaria bicolor. Seedlings of jack pine (Pinus banksiana) grown nonaseptically in an autoclaved soil substrate were given different L. bicolor inoculum treatments. These included (i) a dikaryotic mycelium genotype (D); (ii) D and basidiospores collected from one group of five sporophores (T1); (iii) D and basidiospores collected from 10 sporophores, two from each of five different groups (T5); (iv) T1 alone; (v) T5 alone; and (vi) a noninoculated control. Dikaryotic mycelial inoculum was provided at the time of sowing, while basidiospore inoculum was added at 10 weeks after seed germination. Sporophore formation was induced after 20 weeks of growth, and dikaryotic cultures were isolated from their tissue. Seedlings were harvested, and growth and mycorrhization were assessed. Levels of both were generally lower for T1 treated seedlings, compared with seedlings receiving D, while levels for T5 treated seedlings were intermediate. Sporophore genotype variability was assessed for inoculum treatments by using the isoenzymatic marker leucine aminopeptidase. The greatest genetic variability was seen with the basidiospore treatments T1 and T5, with up to four leucine aminopeptidase patterns per seedling. The mixed treatments D plus T1 and D plus T5 produced most frequently, but not exclusively, the inoculated dikaryon genotype. After isoenzyme results were assessed, variable sporophore isolates of mixed treatments were analyzed with randomly amplified polymorphic DNA and PCR mitochondrial DNA markers to determine if they were formed by dikaryon-monokaryon crosses between the inoculated dikaryon and monosporous mycelia.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7486996 TI - The relationship between viability and intracellular pH in the yeast Saccharomyces cerevisiae. AB - The relationship between viability (cell proliferation activity) and intracellular pH in the yeast Saccharomyces cerevisiae was investigated by using cells that had been deactivated by low-temperature storage, ethanol treatment, or heat treatment. The intracellular pH was measured with a microscopic image processor or a spectrofluorophotometer. At first, the intracellular pH measurements of individual cells were compared with slide culture results by microscopic image processing. A clear correlation existed between the proliferation activity and intracellular pH. Moreover, by spectrofluorophotometry analysis, it was found that there was a relationship between the viability and intracellular pH of brewing yeast under conditions of low external pH (n = 15, r = 0.960, P = 0.001). This relationship was also observed in baker's yeast (n = 13, r = 0.950, P = 0.001). On the other hand, when the fluorescein staining method was used in these experiments, the relationship between viability and staining percentage was not observed. From these results, intracellular pH was found to be a sensitive factor for estimating yeast physiology. The possible role of cell deterioration is also discussed. PMID- 7486998 TI - stcS, a putative P-450 monooxygenase, is required for the conversion of versicolorin A to sterigmatocystin in Aspergillus nidulans. AB - Sterigmatocystin (ST) and aflatoxin are carcinogenic end point metabolites derived from the same biochemical pathway, which is found in several Aspergillus spp. Recently, an ST gene cluster, containing approximately 25 distinct genes that are each proposed to function specifically in ST biosynthesis, has been identified in Aspergillus nidulans. Each of these structural genes is named stc (sterigmatocystin) followed by a consecutive letter of the alphabet. We have previously described stcU (formerly verA) as encoding a keto-reductase required for the conversion of versicolorin A to ST. We now describe a second A. nidulans gene, stcS (formerly verB), that is located within 2 kb of stcU in the ST gene cluster. An stcS-disrupted strain of A. nidulans, TSS17, was unable to produce ST and converted ST/aflatoxin precursors to versicolorin A rather than ST, indicating that stcS functions at the same point in the pathway as stcU. Genomic sequence analysis of stcS shows that it encodes a cytochrome P-450 monooxygenase and constitutes a novel P-450 family, CYP59. Assuming that StcU activity mimics that of similar P-450s, it is likely that StcU catalyzes one of the proposed oxidation steps necessary to convert versicolorin A to ST. These results constitute the first genetic proof that the conversion of versicolorin A to ST requires more than one enzymatic activity. PMID- 7486999 TI - Heat, hydrogen peroxide, and UV resistance of Bacillus subtilis spores with increased core water content and with or without major DNA-binding proteins. AB - Spores of a Bacillus subtilis strain with an insertion mutation in the dacB gene, which codes for an enzyme involved in spore cortex biosynthesis, have a higher core water content than wild-type spores. Spores lacking the two major alpha/beta type small, acid-soluble proteins (SASP) (termed alpha-beta- spores) have the same core water content as do wild-type spores, but alpha-beta- dacB spores had more core water than did dacB spores. The resistance of alpha-beta-, alpha-beta- dacB, dacB, and wild-type spores to dry and moist heat, hydrogen peroxide, and UV radiation has been determined, as has the role of DNA damage in spore killing by moist heat and hydrogen peroxide. These data (i) suggest that core water content has little if any role in spore UV resistance and are consistent with binding of alpha/beta-type SASP to DNA being the major mechanism providing protection to spores from UV radiation; (ii) suggest that binding of alpha/beta-type SASP to DNA is the major mechanism unique to spores providing protection from dry heat; (iii) suggest that spore resistance to moist heat and hydrogen peroxide is affected to a large degree by the core water content, as increased core water resulted in large decreases in spore resistance to these agents; and (iv) indicate that since this decreased resistance (i.e., in dacB spores) is not associated with increased spore killing by DNA damage, spore DNA must normally be extremely well protected against such damage, presumably by the saturation of spore DNA by alpha/beta-type SASP. PMID- 7487000 TI - PCR for bioaerosol monitoring: sensitivity and environmental interference. AB - The PCR technique has potential for use in detection of low concentrations of airborne microorganisms. In this study, the sensitivity of PCR and its susceptibility to environmental interference were assessed with Escherichia coli DH1 as the target organism. Air samples, containing environmental bioaerosols, were collected with AGI-30 samplers and seeded with E. coli DH1 cells. Parallel studies were performed with cells seeded into the sampler prior to collection of air samples to determine the effects of environmental inhibition and sampling stress on the PCR assay. Baseline studies were also performed without environmental challenge or sampling stress to compare two protocols for cell lysis, solid phase and freeze-thaw. Amplification of a plasmid target sequence resulted in a detection limit of a single bacterial cell by the freeze-thaw and solid-phase methods within 5 and 9 h, respectively. With a genomic target, the sensitivity of the solid-phase method was 10-fold lower than that of freeze-thaw. Samples which contained 10(3) to 10(4) CFU of environmental organisms per m3 inhibited amplification; however, a 1/10 dilution of these samples resulted in successful amplifications. No difference in sensitivity of the PCR assay was obtained as a result of sampling stress, although a 10-fold decrease in culturability was observed. A field validation of the protocol with genomic primers demonstrated the presence of airborne E. coli and/or Shigella spp. in outdoor samples. This study indicates that the PCR method for detection of airborne microorganisms is rapid and sensitive and can be used as an alternative method for air quality monitoring. PMID- 7487001 TI - Transformations of morphine alkaloids by Pseudomonas putida M10. AB - The oxidation of morphine by washed-cell incubations of Pseudomonas putida M10 gave rise to a large number of transformation products including hydromorphone (dihydromorphinone), 14 beta-hydroxymorphine, 14 beta-hydroxymorphinone, and dihydromorphine. Similarly, in incubations with oxymorphone (14 beta hydroxydihydromorphinone) as substrate, the major transformation product was identified as oxymorphol (14 beta-hydroxydihydromorphine). The identities of all these biological products were confirmed by mass spectrometry and 1H nuclear magnetic resonance spectroscopy. This is the first report describing structural evidence for the biological synthesis of 14 beta-hydroxymorphine and 14 beta hydroxymorphinone. These products have applications as intermediates in the synthesis of semisynthetic opiate drugs. PMID- 7487002 TI - Production of Trichoderma reesei cellulases on glucose-containing media. AB - The filamentous fungus Trichoderma reesei was shown to secrete active cellobiohydrolase I and the endoglucanase I catalytic core domain into the culture medium when the fungus was grown on glucose-containing medium. The expression of the proteins was driven by the promoters of the elongation factor 1 alpha, tef1, and the unidentified gene for cDNA1. The cDNA1 promoter gave the best yields. The highest amounts of cellobiohydrolase I and the endoglucanase I core, being 50 to 100 mg/liter, accounted for more than half of the total protein secreted by the fungus. The levels obtained with the tef1 promoter were 20 to 50 times lower. PMID- 7487003 TI - Uptake and retention of Vibrio cholerae O1 in the Eastern oyster, Crassostrea virginica. AB - Vibrio cholerae 01, the causative agent of cholera, is known to persist in estuarine environments as endogenous microflora. The recent introduction of V. cholerae 01 into estuaries of the North and South American continents has stimulated the need to determine the effect of controlled purification on reducing this pathogen in edible molluscan shellfish. Experiments defined parameters for the uptake and retention of V. cholerae 01 in tissues of Crassostrea virginica, and these parameters were compared with those for Escherichia coli and Salmonella tallahassee, bacteria which are usually eliminated from moderately contaminated shellfish within 48 h. Oysters accumulated greater concentrations of V. cholerae 01 than E. coli and S. tallahassee. When V. cholerae 01 was exposed to controlled purification at 15, 19 and 25 degrees C over 48 h, it persisted in oysters at markedly higher levels than E. coli and S. tallahassee. The concentration of a V. cholerae 01-specific agglutinin did not positively correlate with the uptake or retention of V. cholerae 01. These data show that state and federally approved controlled purification techniques are not effective at reducing V. cholerae 01 in oysters. PMID- 7487004 TI - A novel approach for monitoring genetically engineered microorganisms by using artificial, stable RNAs. AB - Further improvements in technology for efficient monitoring of genetically engineered microorganisms (GEMs) in the environment are needed. Technology for monitoring rRNA is well established but has not generally been applicable to GEMs because of the lack of unique rRNA target sequences. In the work described herein, it is demonstrated that a deletion mutant of a plasmid-borne Vibrio proteolyticus 5S rRNA gene continues to accumulate to high levels in Escherichia coli although it is no longer incorporated into 70S ribosomes. This deletion construct was subsequently modified by mutagenesis to create a unique recognition site for the restriction endonuclease BstEII, into which new sequences could be readily inserted. Finally, a novel 17-nucleotide identifier sequence from Pennisetum purpureum was embedded into the construct to create an RNA identification cassette. The artificial identifier RNA, expressed from this cassette in vivo, accumulated in E. coli to levels comparable to those of wild type 5S rRNA without being seriously detrimental to cell survival in laboratory experiments and without entering the ribosomes. These results demonstrate that artificial, stable RNAs containing sequence segments remarkably different from those present in any known rRNA can be designed and that neither the deleted sequence segment nor ribosome incorporation is essential for accumulation of an RNA product. PMID- 7487005 TI - Evidence for indigenous Streptomyces populations in a marine environment determined with a 16S rRNA probe. AB - A 16S rRNA genus-specific probe was used to determine whether Streptomyces populations are an indigenous component of marine sediment bacterial communities. Previous debates have suggested that marine Streptomyces isolates are derived not from resident populations but from spores of terrestrial species which have been physically transported to marine ecosystems but remain dormant until isolation. Rigorously controlled hybridization of rRNA extracted from coastal marsh sediments with the genus-specific probe indicated that Streptomyces rRNA accounted for 2 to 5% of the sediment community rRNA and that spores are not the source of the hybridization signal. Streptomyces populations must therefore be at least the 26th most abundant genus-level source of bacterial rRNA. the relative amounts of rRNAs from Streptomyces spp. and members of the Bacteria (69 to 79%) and Archaea (4 to 7%) domains were highly consistent in these marine sediments throughout an annual cycle, indicating that the species composition of sediment bacterial communities may be more stable than recent studies suggest for marine planktonic bacterial communities. Laboratory studies designed to investigate the possible functional roles of Streptomyces populations in coastal sediments demonstrated that population levels of this genus changed relatively rapidly (within a time frame of 6 weeks) in response to manipulation of substrate availability. Amendments of intact sediment cores with two compounds (vanillic acid and succinic acid) consistently resulted in Streptomyces populations contributing an increased percentage of rRNA (6 to 15%) to the total bacterial rRNA pool. PMID- 7487006 TI - Characterization of a keratinolytic serine proteinase from Streptomyces pactum DSM 40530. AB - A serine protease from the keratin-degrading Streptomyces pactum DSM 40530 was purified by casein agarose affinity chromatography. The enzyme had a molecular weight of 30,000 and an isoelectric point of 8.5. The proteinase was optimally active in the pH range from 7 to 10 and at temperatures from 40 to 75 degrees C. The enzyme was specific for arginine and lysine at the P1 site and for phenylalanine and arginine at the P1' site. It showed a high stereoselectivity and secondary specificity with different synthetic substrates. The keratinolytic activity of the purified proteinase was examined by incubation with the insoluble substrates keratin azure, feather meal, and native and autoclaved chicken feather downs. The S. pactum proteinase was significantly more active than the various commercially available proteinases. After incubation with the purified proteinase, a rapid disintegration of whole feathers was observed. But even after several days of incubation with repeated addition of enzymes, less than 10% of the native keratin substrate was solubilized. In the presence of dithiothreitol, degradation was more than 70%. PMID- 7487008 TI - Use of a fluorogenic probe in a PCR-based assay for the detection of Listeria monocytogenes. AB - A PCR-based assay for Listeria monocytogenes that uses the hydrolysis of an internal fluorogenic probe to monitor the amplification of the target has been formatted. The fluorogenic 5' nuclease PCR assay takes advantage of the endogenous 5' --> 3' nuclease activity of Taq DNA polymerase to digest a probe which is labelled with two fluorescent dyes and hybridizes to the amplicon during PCR. When the probe is intact, the two fluorophores interact such that the emission of the reporter dye is quenched. During amplification, the probe is hydrolyzed, relieving the quenching of the reporter and resulting in an increase in its fluorescence intensity. This change in reporter dye fluorescence is quantitative for the amount of PCR product and, under appropriate conditions, for the amount of template. We have applied the fluorogenic 5' nuclease PCR assay to detect L. monocytogenes, using an 858-bp amplicon of hemolysin (hlyA) as the target. Maximum sensitivity was achieved by evaluating various fluorogenic probes and then optimizing the assay components and cycling parameters. With crude cell lysates, the total assay could be completed in 3 h with a detection limit of approximately 50 CFU. Quantification was linear over a range of 5 x 10(1) to 5 x 10(5) CFU. PMID- 7487007 TI - Actions of a versatile fluorene-degrading bacterial isolate on polycyclic aromatic compounds. AB - Pseudomonas cepacia F297 grew with fluorene as a sole source of carbon and energy; its growth yield corresponded to an assimilation of about 40% of fluorene carbon. The accumulation of a ring meta-cleavage product during growth and the identification of 1-indanone in growth media and washed-cell suspensions suggest that strain F297 metabolizes fluorene by mechanisms analogous to those of naphthalene degradation. In addition to fluorene, strain F297 utilized for growth a wide variety of polycyclic aromatic compounds (PACs), including naphthalene, 2,3-dimethylnaphthalene, phenanthrene, anthracene, and dibenzothiophene. Fluorene induced cells of the strain also transformed 2,6-dimethylnaphthalene, biphenyl, dibenzofuran, acenaphthene, and acenaphthylene. The identification of products formed from those substrates (by gas chromatography-mass spectrometry) in washed cell suspensions indicates that P. cepacia F297 carries out the following reactions: (i) aromatic ring oxidation and cleavage, apparently using the pyruvate released for growth, (ii) methyl group oxidations, (iii) methylenic oxidations, and (iv) S oxidations of aromatic sulfur heterocycles. Strain F297 grew with a creosote-PAC mixture, producing an almost complete removal of all aromatic compounds containing 2 to 3 rings in 14 days, as demonstrated by gas chromatography analysis of the remaining PACs recovered from cultures. The identification of key chemicals confirmed that not only are certain compounds depleted but also the anticipated reaction products are found. PMID- 7487009 TI - Substrate-dependent differential splicing of introns in the regions encoding the cellulose binding domains of two exocellobiohydrolase I-like genes in Phanerochaete chrysosporium. AB - Recently, we have shown differential splicing of an intron in the cbhI.2 gene of Phanerochaete chrysosporium ME446; this intron lies within the region of the gene encoding the cellulose binding domain (P.F.G. Sims, M. S. Soares-Felipe, Q. Wang, M.E. Gent, C. Tempelaars, and P. Broda, Mol. Microbiol. 12:209-216, 1994). Here, we show that such differential splicing occurs in the cbhI.1 gene of this fungus as well as in the cbhI.2 gene and that this phenomenon is substrate dependent. Avicel elicits the synthesis of both classes of mRNA transcripts from both of these genes. In contrast, carboxymethyl cellulose predominantly elicits the synthesis of fully spliced transcripts from both genes. Such differential splicing might allow this fungus to regulate the specificities of substrate binding for these cellulases. PMID- 7487010 TI - A new method for direct detection of Listeria monocytogenes from foods by PCR. AB - The preparation of DNA by alcohol precipitation in the presence of Na1 was used to enable the direct detection of Listeria monocytogenes in the amount of 10(3) CFU/0.5 g of sample. This procedure produces PCR-quality DNA directly from foods, such as soft cheese and minced meat. PMID- 7487011 TI - Acid adaptation of Escherichia coli O157:H7 increases survival in acidic foods. AB - Escherichia coli O157:H7 was adapted to acid by culturing for one to two doublings at pH 5.0. Acid-adapted cells had an increased resistance to lactic acid, survived better than nonadapted cells during a sausage fermentation, and showed enhanced survival in shredded dry salami (pH 5.0) and apple cider (pH 3.4). Acid adaptation is important for the survival of E. coli O157:H7 in acidic foods and should be considered a prerequisite for inocula used in food challenge studies. PMID- 7487012 TI - Efficacy of the Ryu nonstaining KOH technique for rapidly determining gram reactions of food-borne and waterborne bacteria and yeasts. AB - A simple and rapid (< 60 s) nonstaining technique with 3% potassium hydroxide to determine Gram reactions was tested with 495 food-borne and waterborne bacteria and yeasts. In KOH, suspensions of gram-negative bacteria become viscous and string out. Gram-positive bacteria are not affected. There was 100% correlation between the KOH string test results and gram-positive and gram-negative strains. PMID- 7487013 TI - Identification of algae which interfere with the detection of Giardia cysts and Cryptosporidium oocysts and a method for alleviating this interference. AB - Fifty-four algal species were tested for cross-reaction in the American Society for Testing and Materials Giardia/Cryptosporidium indirect immunofluorescence assay, and 24 showed some degree of fluorescence. Two species, Navicula minima and Synechococcus elongatus, exhibited a bright apple green fluorescence. The addition of goat serum to the assay mixture blocked the fluorescence of most nontarget organisms tested and also decreased the background fluorescence. Goat serum did not interfere with the fluorescence of Giardia cysts or Cryptosporidium oocysts or the identification of cyst and oocyst internal structures. PMID- 7487014 TI - Development of a PCR for identification of Naegleria fowleri from the environment. AB - A species-specific PCR for the identification of Naegleria fowleri was developed. In sensitivity studies, 10 trophozoites or cysts and 1 trophozoite or cyst could be detected after 35 and 45 cycles, respectively. In conjunction with a rapid DNA isolation method, this PCR was used to identify N. fowleri directly from primary cultures of environmental samples. PMID- 7487015 TI - Behavior of Escherichia coli and male-specific bacteriophage in environmentally contaminated bivalve molluscs before and after depuration. AB - We monitored the differential reduction rates and elimination patterns of Escherichia coli and male-specific (F+) bacteriophage during UV depuration for 48 h in oysters (Crassostrea gigas) and mussels (Mytilus edulis) contaminated by short-term (1 to 3 weeks) and long-term (more than 6 months) exposure to sewage in the marine environment. The time taken to reduce levels of E. coli by 90% was 6.5 h or less in all cases. In contrast, the amounts of time needed to reduce levels of F+ bacteriophage by 90% were considerably longer: 47.3 and 41.3 h (after short- and long-term exposures, respectively) in mussels and 54.6 and 60.8 h (after short- and long-term exposures, respectively) in oysters. No differences in the rates of reduction of indicators of viral pollution following exposure of the shellfish to either short- or long-term sewage contamination were observed. Further experiments were conducted with mussels to determine the relative distributions of E. coli and F+ bacteriophage in tissue before and during depuration. Prior to depuration the majority of E. coli organisms (90.1%) and F+ bacteriophage (87.3%) were detected in the digestive tract (i.e., the digestive gland and intestine). E. coli and F+ bacteriophage were reduced in all tissues except the digestive gland to undetectable levels following depuration for 48 h. Within the digestive gland, levels of F+ bacteriophage were reduced to 30% of initial levels, whereas E. coli was reduced to undetectable levels.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7487016 TI - Characterization of lipopolysaccharide heterogeneity in Salmonella enteritidis by an improved gel electrophoresis method. AB - Salmonella enteritidis field isolates of different phage types and pathogenicities were assessed for changes in lipopolysaccharide (LPS) structure, using an improved method of polyacrylamide gel electrophoresis (PAGE) that revealed the same degree of structural detail as mass spectroscopy. The method allowed characterization of an LPS chemotype that may be associated, regardless of phage type, with increased virulence of S. enteritidis. The virulent variant SE6-E21, which efficiently contaminates eggs and yields high numbers of organisms from chick spleens, had an O-antigen/core ratio of 2.8, as determined from gels by densitometry, and 1.67 micrograms of mannose per microgram of 2-keto-3-deoxy octulosonic acid (KDO), while the avirulent variant SE6-E5 had O-antigen/core ratios of 1.2 and 1.00. The association between O antigen and virulence was also seen on analysis of five new field isolates. One of the new field isolates generated a mixed population of smooth and semismooth variants in agreement with its mixed virulence in chicks. When LPS was purified from large-volume cultures, only the most virulent isolate yielded high amounts of O antigen (1.6 micrograms of mannose per microgram of KDO), while the other isolates had ratios characteristic of semismooth variants (< or = 1.0 microgram of mannose per microgram of KDO), including the isolate of mixed virulence. These results indicate that the improved PAGE method might provide a rapid, sensitive, in vitro assessment of field isolate virulence prior to the performance of definitive infectivity trials. PMID- 7487018 TI - Role of competition for inorganic nutrients in the biodegradation of mixtures of substrates. AB - A study was conducted to determine whether competition for inorganic nutrients affects the biodegradation of mixtures of substrates. Little benzylamine was mineralized by Pseudomonas putida in solutions with no added P, but the substrate was degraded if the medium contained 100 nM P. The enhancement by P addition did not occur if the medium also contained caprolactam and a caprolactam-utilizing strain of Pseudomonas aeruginosa. The suppression by the second bacterium was overcome by a higher P concentration. The rate of caprolactam utilization by P. aeruginosa was reduced if benzylamine and P. putida were also present in media with 100 nM P, but the suppression was absent if the solution contained a higher P concentration. Glutamate increased and inorganic N plus P decreased the length of the acclimation phase prior to benzylamine mineralization in lake water. We suggest that the effect of one biodegradable substrate on the metabolism of a second often results from a competition for inorganic nutrients. PMID- 7487017 TI - Soil and sediment bacteria capable of aerobic nitrate respiration. AB - Several laboratory strains of gram-negative bacteria are known to be able to respire nitrate in the presence of oxygen, although the physiological advantage gained from this process is not entirely clear. The contribution that aerobic nitrate respiration makes to the environmental nitrogen cycle has not been studied. As a first step in addressing this question, a strategy which allows for the isolation of organisms capable of reducing nitrate to nitrite following aerobic growth has been developed. Twenty-nine such strains have been isolated from three soils and a freshwater sediment and shown to comprise members of three genera (Pseudomonas, Aeromonas, and Moraxella). All of these strains expressed a nitrate reductase with an active site located in the periplasmic compartment. Twenty-two of the strains showed significant rates of nitrate respiration in the presence of oxygen when assayed with physiological electron donors. Also isolated was one member of the gram-positive genus Arthrobacter, which was likewise able to respire nitrate in the presence of oxygen but appeared to express a different type of nitrate reductase. In the four environments studied, culturable bacteria capable of aerobic nitrate respiration were isolated in significant numbers (10(4) to 10(7) per g of soil or sediment) and in three cases were as abundant as, or more abundant than, culturable bacteria capable of denitrification. Thus, it seems likely that the corespiration of nitrate and oxygen may indeed make a significant contribution to the flux of nitrate to nitrite in the environment. PMID- 7487019 TI - Improvement of solubility and stability of the antimicrobial peptide nisin by protein engineering. AB - Nisin is a 3.4-kDa antimicrobial peptide that, as a result of posttranslational modifications, contains unsaturated amino acids and lanthionine residues. It is applied as a preservative in various food products. The solubility and stability of nisin and nisin mutants have been studied. It is demonstrated that nisin mutants can be produced with improved functional properties. The solubility of nisin A is highest at low pH values and gradually decreases by almost 2 orders of magnitude when the pH of the solution exceeds a value of 7. At low pH, nisin Z exhibits a decreased solubility relative to that of nisin A; at neutral and higher pH values, the solubilities of both variants are comparable. Two mutants of nisin Z, which contain lysyl residues at positions 27 and 31, respectively, instead of Asn-27 and His-31, were produced with the aim of reaching higher solubility at neutral pH. Both mutants were purified to homogeneity, and their structures were confirmed by one- and two-dimensional 1H nuclear magnetic resonance. Their antimicrobial activities were found to be similar to that of nisin Z, whereas their solubilities at pH 7 increased by factors of 4 and 7, respectively. The chemical stability of nisin A was studied in the pH range of 2 to 8 and at a 20, 37, and 75 degrees C. Optimal stability was observed at pH 3.0. Nisin Z showed a behavior similar to that of nisin A. A mutant containing dehydrobutyrine at position 5 instead of dehydroalanine had lower activity but was significantly more resistant to acid-catalyzed chemical degradation than wild type nisin Z. PMID- 7487021 TI - Functional analysis of the threonine- and serine-rich Gp-I domain of glucoamylase I from Aspergillus awamori var. kawachi. AB - Glucoamylase I (GAI) from Aspergillus awamori var. kawachi hydrolyzes raw starch efficiently and is composed of three functional domains: the amino-terminal catalytic GAI' domain (A-1 to V-469), the threonine- and serine-rich O glycosylated Gp-I domain (A-470 to V-514), and the carboxy-terminal raw starch binding Cp domain (A-515 to R-615). In order to investigate the role of the Gp-I domain, an additional repeat of Gp-I and internal deletions of the entire Gp-I sequence or parts of the Gp-I sequence were introduced within Gp-I. All mutant genes as well as the wild-type gene were inserted into a yeast-secretion vector, YEUp3H alpha, and expressed in Saccharomyces cerevisiae. Wild-type GAI expressed in yeast cells (GAY), GAGpI, having an extra Gp-I, and GA delta 470-493, lacking the A-470-to-T-493 sequences of Gp-I, were successfully secreted into the culture medium. On the other hand, GA delta 470-507, lacking A-470 to S-507, and GA delta GpI, lacking the entire Gp-I (A-470-to-V-514) sequence, failed to be secreted and remained in the yeast cells. The carbohydrate content of GAGpI was 1.2 times higher than that of GAY and 2.4 times higher than that of the original GAI. The raw starch digestibility of GAGpI was almost the same as that of GAY but was 1.5 times faster than that of GAI.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7487020 TI - Universal PCR primers for detection of phytopathogenic Agrobacterium strains. AB - Two PCR primer pairs, based on the virD2 and ipt genes, detected a wide variety of pathogenic Agrobacterium strains. The endonuclease domain of VirD2 protein, which cleaves transferred DNA (T-DNA) border sequences, is highly conserved; primer oligonucleotides specific for the endonuclease portion of virD2 detected all pathogenic strains of Agrobacterium tested. PCR primers corresponding to conserved sequences in ipt, the T-DNA-borne cytokinin synthesis gene, detected only Agrobacterium tumefaciens and distinguished it from Agrobacterium rhizogenes. The virD2 and ipt primer pairs did not interfere with each other when included in the same PCR amplification, and this permitted simultaneous detection of both genes in a single reaction. One nonpathogenic Agrobacterium radiobacter strain contained virD2 but not ipt; we speculate that this strain arose from a pathogenic progenitor through a deletion in the T-DNA. The virD2 primer pair appears to be universal for all pathogenic Agrobacterium species; used together, the primer sets reported here should allow unambiguous identification of Ti plasmid DNA in bacteria isolated from soil and plants. PMID- 7487023 TI - Enterobacterial repetitive intergenic consensus sequences and the PCR to generate fingerprints of genomic DNAs from Vibrio cholerae O1, O139, and non-O1 strains. AB - Enterobacterial repetitive intergenic consensus (ERIC) sequence polymorphism was studied in Vibrio Cholerae strains isolated before and after the cholera epidemic in Brazil (in 1991), along with epidemic strains from Peru, Mexico, and India, by PCR. A total of 17 fingerprint patterns (FPs) were detected in the V. cholerae strains examined; 96.7% of the toxigenic V. cholerae O1 strains and 100% of the O139 serogroup strains were found to belong to the same FP group comprising four fragments (FP1). The nontoxigenic V. cholerae O1 also yielded four fragments but constituted a different FP group (FP2). A total of 15 different patterns were observed among the V. cholerae non-O1 strains. Two patterns were observed most frequently for V. cholerae non-01 strains, 25% of which have FP3, with five fragments, and 16.7% of which have FP4, with two fragments. Three fragments, 1.75, 0.79, and 0.5 kb, were found to be common to both toxigenic and nontoxigenic V. cholerae O1 strains as well as to group FP3, containing V. cholerae non-O1 strains. Two fragments of group FP3, 1.3 and 1.0 kb, were present in FP1 and FP2 respectively. The 0.5-kb fragment was common to all strains and serogroups of V. cholerae analyzed. It is concluded from the results of this study, based on DNA FPs of environmental isolates, that it is possible to detect an emerging virulent strain in a cholera-endemic region. ERIC-PCR constitutes a powerful tool for determination of the virulence potential of V. cholerae O1 strains isolated in surveillance programs and for molecular epidemiological investigations. PMID- 7487024 TI - Identification of Clostridium tyrobutyricum as the causative agent of late blowing in cheese by species-specific PCR amplification. AB - Butyric acid fermentation, the late-blowing defect in cheese, caused by the outgrowth of clostridial spores present in raw milk, can create considerable loss of product, especially in the production of semihard cheeses like Gouda cheese, but also in grana and Gruyere cheeses. To demonstrate the causative relationship between Clostridium tyrobutyricum and late blowing in cheese, many cheesemaking experiments were performed to provoke this defect by using spores from several strains of the major dairy-related clostridia. A method of PCR amplification of a part of the 16S rRNA gene in combination with hybridization with species-specific DNA probes was developed to allow the specific detection of clostridial sequences in DNAs extracted from cheeses. The sensitivity was increased by using nested PCR. Late blowing was provoked in experimental cheeses with 28 of the 32 C. tyrobutyricum strains tested, whereas experimental cheeses made with spores from C. beijerinckii, C. butyricum, and C. sporogenes showed no signs of butyric acid fermentation. In all experimental and commercial cheeses with obvious signs of late blowing, DNA from C. tyrobutyricum was detected; in some cheeses, signals for C. beijerinckii were also found. It was concluded that only C. tyrobutyricum strains are able to cause butyric acid fermentation in cheese. PMID- 7487025 TI - Comparison of different procedures for serotyping aquatic birnavirus. AB - The current classification of aquatic birnaviruses is based on seroneutralization assays with polyclonal antibodies. In this study a comparison of several procedures used for serotyping aquatic birnaviruses was made with 10 virus strains (4 reference strains from salmonids and 6 birnaviruses isolated from turbot [Scophthalmus maximus]). The relationships among the birnavirus strains were studied by seroneutralization assay with polyclonal antibodies and by immunodot assay with both polyclonal and monoclonal antibodies. The results were compared with a presumptive classification obtained from analysis of restriction enzyme patterns of cDNA products obtained by PCR amplification. No correlation was found among the results obtained by the different procedures. The seroneutralization and the immunodot assays with polyclonal antibodies were not useful in classifying these birnaviruses strains; however, patterns of reaction with monoclonal antibodies emphasized the individuality of the strains, particularly in the case of two strains (231 and 460) whose patterns did not correspond to established serotypes. The application of PCR and restriction enzyme analysis is a promising system for approaching the classification of this viral group on the basis of genomic differences and similarities. The variable results obtained in this comparison lead us to think that the current classification of aquatic birnavirus may not be the most accurate and there is a need for modification incorporating recent isolates, not only from salmonid species but also from marine fish. PMID- 7487022 TI - Expression of the strA-strB streptomycin resistance genes in Pseudomonas syringae and Xanthomonas campestris and characterization of IS6100 in X. campestris. AB - Expression of the strA-strB streptomycin resistance (SMr) genes was examined in Pseudomonas syringae pv. syringae and Xanthomonas campestris pv. vesicatoria. The strA-strB genes in P. syringae and X. campestris were encoded on elements closely related to Tn5393 from Erwinia amylovora and designated Tn5393a and Tn5393b, respectively. The putative recombination site (res) and resolvase-repressor (tnpR) genes of Tn5393 from E. amylovora, P syringae, and X. campestris were identical; however, IS6100 mapped within tnpR in X. campestris, and IS1133 was previously located downstream of tnpR in E. amylovora (C.-S Chiou and A. L. Jones, J. Bacteriol. 175:732-740, 1993). Transcriptional fusions (strA strB::uidA) indicated that a strong promoter sequence was located within res in Tn5393a. Expression from this promoter sequence was reduced when the tnpR gene was present in cis position relative to the promoter. In X. campestris pv. vesicatoria, analysis of promoter activity with transcriptional fusions indicated that IS6100 increased the expression of strA-strB. Analysis of codon usage patterns and percent G+C in the third codon position indicated that IS6100 could have originated in a gram-negative bacterium. The data obtained in the present study help explain differences observed in the levels of SMr expressed by three genera which share common genes for resistance. Furthermore, the widespread dissemination of Tn5393 and derivatives in phytopathogenic prokaryotes confirms the importance of these bacteria as reservoirs of antibiotic resistance in the environment. PMID- 7487026 TI - Continuous degradation of trichloroethylene by Xanthobacter sp. strain Py2 during growth on propene. AB - Propene-grown Xanthobacter sp. strain Py2 cells can degrade trichloroethylene (TCE), but the transformation capacity of such cells was limited and depended on both the TCE concentration and the biomass concentration. Toxic metabolites presumably accumulated extracellularly, because the fermentation of glucose by yeast cells was inhibited by TCE degradation products formed by strain Py2. The affinity of the propene monooxygenase for TCE was low, and this allowed strain Py2 to grow on propene in the presence of TCE. During batch growth with propene and TCE, the TCE was not degraded before most of the propene had been consumed. Continuous degradation of TCE in a chemostat culture of strain Py2 growing with propene was observed with TCE concentrations up to 206 microns in the growth medium without washout of the fermentor occurring. At this TCE concentration the specific degradation rate was 1.5 nmol/min/mg of biomass. The total amount of TCE that could be degraded during simultaneous growth on propene depended on the TCE concentration and ranged from 0.03 to 0.34g of TCE per g of biomass. The biomass yield on propene was not affected by the cometabolic degradation of TCE. PMID- 7487027 TI - Characterization of rhizosphere colonization by luminescent Enterobacter cloacae at the population and single-cell levels. AB - A bioluminescence marker system was used to characterized colonization of the rhizosphere by a bacterial inoculum, both in terms of population activity and at the single-cell level. Plasmid pQF70/44, which contains luxAB genes under the control of a strong constitutive phage promoter, was introduced into the rhizobacterium and model biocontrol agent Enterobacter cloacae. Light output from the lux-modified strain was detected by luminometry of samples from growing cultures of E. cloacae and from inoculated soil and wheat root samples. The minimum detection limits for fully active cells under optimum conditions were 90 and 445 cells g-1 for liquid culture and soil, respectively. The metabolic activities of the lux-marked population of E. cloacae, characterized by luminometry, contrasted in rhizosphere and nonrhizosphere soil. Cells in the rhizosphere were active, and there was a linear relationship between light output and cell concentration. The activity of cells in nonrhizosphere coil could not be detected unless the soil was supplied with substrate. Novel use of a charge coupled device is reported for the spatial characterization of rhizosphere colonization by E. cloacae (pQF70/44) at the single-cell and population levels. Used macroscopically, the charge-coupled device identified differences in colonization due to competition from indigenous soil organisms. The lux-marked bacterium was able to colonize all depths of roots in the absence of competition but was restricted tot he spermosphere in the presence of competition (nonsterile soil).(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7487029 TI - Isolation and Identification of novel toxins from a new mosquitocidal isolate from Malaysia, Bacillus thuringiensis subsp. jegathesan. AB - A new mosquitocidal Bacillus thuringiensis subsp., jegathesan, has recently been isolated from Malaysia. Parasporal crystal inclusions were purified from this strain and bioassayed against fourth-instar larvae of Culex quinquefasciatus, Aedes aegypti, Aedes togoi, Aedes albopictus, Anopheles maculatus, and Mansonia uniformis. The 50% lethal concentration of crystal inclusions for each species was 0.34, 8.08, 0.34, 17.59, 3.91, and 120 ng/ml, respectively. These values show that parasporal inclusions from this new subspecies have mosquitocidal toxicity comparable to that of inclusions isolated from B. thuringiensis subsp. israelensis. Solubilized and chymotrypsin-activated parasporal inclusions possessed low-level hemolytic activity. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis revealed that the crystals were composed of polypeptides of 77, 74, 72, 68, 55, 38, 35, 27, and 23 kDa. Analysis by Western blotting (immunoblotting) with polyclonal antisera raised against toxins purified from B. thuringiensis subsp. israelensis reveals that proteins in parasporal inclusions of subsp. jegathesan are distinct, because little cross-reactivity was shown. Analysis of the plasmid content of B. thuringiensis subsp. jegathesan indicates that the genes for toxin production may be located on 105- to 120-kb plasmids. Cry- clones that have been cured of these plasmids are nontoxic. Southern blot analysis of plasmid and chromosomal DNA from subsp. jegathesan showed little or low homology to the genes coding for CryIVA, CryIVB, and CryIVD from B. thuringiensis subsp. israelensis. PMID- 7487030 TI - Construction and behavior of biologically contained bacteria for environmental applications in bioremediation. AB - The survival of microorganisms can be predicted through the use of active biological containment systems. We have constructed contained Pseudomonas putida strains that degrade alkylbenzoates. The modified strain carries a fusion of the Plac promoter to the gef gene, which encodes a killing protein. Expression from Plac is controlled through a regulatory cascade, so that Plac is switched on or off by the absence or presence of alkylbenzoates, respectively. Similar uncontained strains were also constructed and tested as a control. Contained and uncontained strains were genetically stable, and their survival and functionality in soil microcosms were as expected. Both contained and uncontained strains survived well in soils supplemented with alkylaromatics, whereas survival of the contained strain in soil microcosms without methylbenzoates was markedly reduced, in contrast to the control strain, which survived in these soils in the absence of alkylbenzoates. The TOL plasmid was transferred in soils between Pseudomonas strains but was not able to mobilize the elements of the containment system. PMID- 7487028 TI - A xylan hydrolase gene cluster in Prevotella ruminicola B(1)4: sequence relationships, synergistic interactions, and oxygen sensitivity of a novel enzyme with exoxylanase and beta-(1,4)-xylosidase activities. AB - Two genes concerned with xylan degradation were found to be closely linked in the ruminal anaerobe Prevotella ruminicola B(1)4, being separated by an intergenic region of 75 nucleotides. xynA is shown to encode a family F endoxylanase of 369 amino acids, including a putative amino-terminal signal peptide. xynB encodes an enzyme of 319 amino acids, with no obvious signal peptide, that shows 68% amino acid identity with the xsa product of Bacteroides ovatus and 31% amino acid identity with a beta-xylosidase from Clostridium stercorarium; together, these three enzymes define a new family of beta-(1,4)-glycosidases. The activity of the cloned P. ruminicola xynB gene product, but not that of the xynA gene product, shows considerable sensitivity to oxygen. Studied under anaerobic conditions, the XynB enzyme was found to act as an exoxylanase, releasing xylose from substrates including xylobiose, xylopentaose, and birch wood xylan, but was relatively inactive against oat spelt xylan. A high degree of synergy (up to 10-fold stimulation) was found with respect to the release of reducing sugars from oat spelt xylan when XynB was combined with the XynA endoxylanase from P. ruminicola B(1)4 or with endoxylanases from the cellulolytic rumen anaerobe Ruminococcus flavefaciens 17. Pretreatment with a fungal arabinofuranosidase also stimulated reducing-sugar release from xylans by XynB. In P. ruminicola the XynA and XynB enzymes may act sequentially in the breakdown of xylan. PMID- 7487031 TI - Bacteriolytic activity caused by the presence of a novel lactococcal plasmid encoding lactococcins A, B, and M. AB - Lactococcus lactis subsp. lactis biovar diacetylactis DPC938 was identified as a bacteriocin-producing strain which exhibited a bacteriolytic effect on other lactococci. Lysis of such target strains was associated with decreases in optical density and release of the intracellular enzyme lactate dehydrogenase. DPC938 exhibits cross-immunity to L. lactis subsp. cremoris 9B4 (M.J. van Belkum, B.J. Hayema, A. Geis, J. Kok, and G. Venema, Appl. Environ. Microbiol. 55:1187-1191, 1989), a strain which produces the bacteriocins lactococcins A, B, and M. Genetic analyses revealed that a 15.5-kb region of DNA encoding these bacteriocins is highly conserved in 9B4, DPC938, and DPC3286, an overproducing derivative of DPC938. This region is located on a 72- and a 78-kb nonmobilizable plasmid in DPC938 and DPC3286, respectively. The bacteriolytic effect exhibited by DPC938 and DPC3286 on sensitive cultures is most probably due to the concerted action of all three bacteriocins. Since these cultures exhibit a lytic effect on lactococci, they have a potential application in the dairy industry as accelerators of starter lysis and hence accelerators of cheese ripening. PMID- 7487033 TI - Molecular characterization of the afl-1 locus in Aspergillus flavus. AB - An unusual mutation at the afl-1 locus, affecting aflatoxin biosynthesis in Aspergillus flavus 649, was investigated. The inability of strain 649 to produce aflatoxin was found to be the result of a large (greater than 60 kb) deletion that included a cluster of aflatoxin biosynthesis genes. Diploids formed by parasexual crosses between strain 649 and the aflatoxigenic strain 86 did not produce aflatoxin, indicating the dominant nature of the afl-1 mutation in strain 649. In metabolite feeding experiments, the diploids did not convert three intermediates in the aflatoxin pathway to aflatoxin. Northern (RNA blot) analysis of the diploids grown in medium conducive for aflatoxin production indicated that the aflatoxin pathway genes nor1, ver1, and omt1 were not expressed; however, there was low-level expression of the regulatory gene aflR. Pulsed-field electrophoresis gels indicated a larger (6 Mb) chromosome in strain 649 than the apparently homologous (4.9 Mb) chromosome in strain 86. The larger chromosome in strain 649 suggests that a rearrangement occurred in addition to the deletion. From these data, we proposed that a trans-sensing mechanism in diploids is responsible for the dominant phenotype associated with the afl-1 locus in strain 649. Such a mechanism is known in Drosophila melanogaster but has not been described for fungi. PMID- 7487034 TI - Oligopeptides are the main source of nitrogen for Lactococcus lactis during growth in milk. AB - The consumption of amino acids and peptides was monitored during growth in milk of proteinase-positive (Prt+) and -negative (Prt-) strains of Lactococcus lactis. The Prt- strains showed monophasic exponential growth, while the Prt+ strains grew in two phases. The first growth phases of the Prt+ and Prt- strains were in same, and no hydrolysis of casein was observed. Also, the levels of consumption of amino acids and peptides in the Prt+ and Prt- strains were similar. At the end of this growth phase, not all free amino acids and peptides were used, indicating that the remaining free amino acids and peptides were unable to sustain growth. The consumption of free amino acids was very low (about 5 mg/liter), suggesting that these nitrogen sources play only a minor role in growth. Oligopeptide transport-deficient strains (Opp-) of L. lactis were unable to utilize oligopeptides and grew poorly in milk. However, a di- and tripeptide transport deficient strain (DtpT-) grew exactly like the wild type (Opp+ Dtpt+) did. These observations indicate that oligopeptides represent the main nitrogen source for growth in milk during the first growth phase. In the second phase of growth of Prt+ strains, milk proteins are hydrolyzed to peptides by the proteinase. Several of the oligopeptides formed are taken up and hydrolyzed internally by peptidases to amino acids, several of which are subsequently released into the medium (see also E.R.S. Kunji, A. Hagting, C.J. De Vries, V. Juillard, A.J. Haandrikman, B. Poolman, and W.N. Konings, J. Biol. Chem. 270:1569-1574, 1995).(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7487032 TI - Detection of Norwalk virus and hepatitis A virus in shellfish tissues with the PCR. AB - A method for the detection of Norwalk virus and hepatitis A virus from shellfish tissues by PCR was developed. Virus was added to the stomach and hepatopancreatic tissues of oysters or hard-shell clams, and viral nucleic acids were purified by a modification of a previously described method (R.L. Atmar, T.G. Metcalf, F.H. Neill, and M.K. Estes, Appl. Environ. Microbiol. 59:631-635, 1993). The new method had the following advantages compared with the previously described method: (i) more rapid sample processing; (ii) increased test sensitivity; (iii) decreased sample-associated interference with reverse transcription-PCR; and (iv) use of chloroform-butanol in place of the chlorofluorocarbon trichlorotrifluoroethane. In addition, internal standards for both Norwalk virus and hepatitis A virus were made which demonstrated when inhibitors to reverse transcription-PCR were present and allowed quantitation of the viral nucleic acids present in samples. This assay can be used to investigate shellfish associated gastroenteritis outbreaks and to study factors involved in virus persistence in shellfish. PMID- 7487035 TI - Characterization of a metalloprotease inhibitor protein (SmaPI) of Serratia marcescens. AB - As suggested by Y. Suh and M.J. Benedik (J. Bacteriol. 174: 2361-2366, 1992), Serratia marcescens ATCC 27117 produced very small amounts (0.8 U ml-1) of an inhibitor protein (SmaPI) that shows an inhibitory activity against extracellular 50-kDa metalloprotease (SMP) of S. marcescens and that is localized in the periplasm of cells at the optimal growth temperature of 25 degrees C. A recombinant S. marcescens harboring plasmid pSP2 encoding SMP and SmaPI genes produced 20 U of SmaPI ml-1 that is also localized in the periplasm of cells at 25 degrees C. However, a large amount of SmaPI (86 Uml-1) was extracellularly produced at the supraoptimal growth temperature 37 degrees C from the recombinant S. marcescens (pSP2). We purified SmaPI from the culture supernatant of S. marcescens (pSP2) grown at 37 degrees C, and some biochemical properties were characterized. SmaPI had a pI value of about 10.0 and was a monomeric protein with a molecular mass of 10,000. SmaPI was produced from a precursor SmaPI by cleavage of a signal peptide (26 amino acid residues). The inhibitor was stable in boiling water for up to 30 min. The thermostability of SmaPI can be attributed to its reversible denaturation. SmaPI inhibited SMP by formation of a noncovalent complex with a molar ratio of 1:1 and showed a high protease specificity, which inhibited only SMP among the various proteases we examined. PMID- 7487036 TI - Degradation and utilization by Butyrivibrio fibrisolvens H17c of xylans with different chemical and physical properties. AB - Hemicelluloses, mainly xylans, can be a major component of diets consumed by ruminants and undergo various degrees of microbial digestion in the rumen. The ability of Butyrivibrio fibrisolvens, a major xylanolytic ruminal species, to degrade and utilize nine chemically and physically different xylans for growth was examined. The arabinoxylans used included two isolated from corncobs (CCX-A and CCX-B), a native xylan excreted by corn cell tissue cultures (CX), an oxalic acid-treated, arabinose-depleted CX, and oat spelt xylan. Except for CCX-A, these xylans were extensively converted within 3 h of growth to acid-alcohol-soluble forms that remained at high levels for the duration of culture growth. These xylans contain mainly xylose and arabinose with small amounts of uronic acids. For a given xylan, all three components were used at about the same rate and extent. During the early stages of growth B. fibrisolvens also rapidly solubilized glucuronoxylans from birchwood, larchwood, 4-O-methylglucuronoxylan, and the xylose homopolymer xylan isolated from beechwood (BEWX). In contrast to the findings for the arabinoxylans, little acid-alcohol-soluble carbohydrate remained in these cultures after 9 h of growth, except for BEWX. Initially, with birchwood, larchwood, and 4-O-methylglucuronoxylan the uronic acid components were preferentially used over the xylose. Final xylan utilization measured at 72 h for all xylans varied from 57% for CCX-A to 92% for BEWX and was correlated with the initial 12-h utilization rate for a given xylan. Since CCX-A and BEWX are both highly water insoluble, this aspect did not appear to influence overall utilization.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7487038 TI - Effect of manganese on preferential degradation of lignin by Pleurotus ostreatus during solid-state fermentation. AB - Practical utilization of the polysaccharides in the lignocellulosic complex is limited because of the high lignin content of the complex. In this study we focused on the effect of Mn on lignin and cellulose biodegradation during solid state fermentation by the edible mushroom Pleurotus ostreatus. Preferential degradation of lignin was enhanced by the addition of Mn(II) to cotton stalks at concentrations ranging from 30 to 620 micrograms of Mn per g. This effect was most apparent when we compared mineralization rates of [14C] lignin with mineralization rates of [14C] cellulose. Enhanced selectivity was also observed when we analyzed residual organic matter at the end of the fermentation period by using crude fiber analysis. The cellulose fraction in the original material was 1.8 times larger than the cellulose fraction of lignin. The cellulose/lignin ratio increased during 32 days of solid-state fermentation from 2.5 in the control to 3.3 following the addition of Mn to the medium. The in vitro digestibility value for fermented cotton stalks was 53% of the dry matter. Addition of 600 micrograms of Mn per g to the cotton stalks resulted in a digestibility value of 65.4%. Enhancement of preferential lignin degradation could be result of either increased activity of the ligninolytic enzymes or production of Mn (III), which might preferentially degrade aromatic structures in the lignocellulosic complex. PMID- 7487037 TI - Abundance of Tn3, Tn21, and Tn501 transposase (tnpA) sequences in bacterial community DNA from marine environments. AB - The occurrence of the tnpA genes of the transposons Tn3, Tn21, and Tn501 was assessed in total bacterial community DNA isolated from different marine environments. The PCR technique was employed, together with most probable number statistics, to determine the abundance of the target tnpA genes. All three genes could be detected, and the Tn21 tnpA sequences predominated in all samples. The smallest amount of total community DNA in which the Tn21 tnpA sequence could be detected was 0.037 ng, and on the basis of our results, we estimated that this sequence was present in 1 of 1,000 to 10,000 bacteria. Hybridization of the PCR products with the respective tnpA probes verified the Tn21 and Tn501 tnpA sequences but only some of the Tn3 tnpA amplification products. The distribution and dissemination of transposons in natural bacterial communities are discussed. PMID- 7487039 TI - Enhanced production of insecticidal proteins in Bacillus thuringiensis strains carrying an additional crystal protein gene in their chromosomes. AB - A two-step procedure was used to place a cryIC crystal protein gene from Bacillus thuringiensis subsp. aizawai into the chromosomes of two B. thuringiensis subsp. kurstaki strains containing multiple crystal protein genes. The B. thuringiensis aizawai cryIC gene, which encodes an insecticidal protein highly specific to Spodoptera exigua (beet armyworm), has not been found in any B. thuringiensis subsp. kurstaki strains. The cryIC gene was cloned into an integration vector which contained a B. thuringiensis chromosomal fragment encoding a phosphatidylinositol-specific phospholipase C, allowing the B. thuringiensis subsp. aizawai cryIC to be targeted to the homologous region of the B. thuringiensis subsp. kurstaki chromosome. First, to minimize the possibility of homologous recombination between cryIC and the resident crystal protein genes, B. thuringiensis subsp. kurstaki HD73, which contained only one crystal gene, was chosen as a recipient and transformed by electroporation. Second, a generalized transducing bacteriophage, CP-51, was used to transfer the integrated cryIC gene from HD73 to two other B. thuringiensis subsp. kurstaki stains. The integrated cryIC gene was expressed at a significant level in all three host strains, and the expression of cryIC did not appear to reduce the expression of the endogenous crystal protein genes. Because of the newly acquired ability to produce the CryIC protein, the recombinant strains showed a higher level of activity against S. exigua than did the parent strains. This two-step procedure should therefore be generally useful for the introduction of an additional crystal protein gene into B. thuringiensis strains which have multiple crystal protein genes and which show a low level of transformation efficiency. PMID- 7487040 TI - Quantitative fluorescence in situ hybridization of Bifidobacterium spp. with genus-specific 16S rRNA-targeted probes and its application in fecal samples. AB - Three 16S rRNA hybridization probes were developed and tested for genus-specific detection of Bifidobacterium species in the human fecal flora. Variable regions V2, V4, and V8 of the 16S rRNA contained sequences unique to this genus and proved applicable as target sites for oligodeoxynucleotide probes. Determination of the genus specificity of the oligonucleotides was performed by whole-cell hybridization with fluorescein isothiocyanate-labelled probes. To this end, cells were fixed on glass slides, hybridized with the probes, and monitored by videomicroscopy. In combination with image analysis, this allowed quantification of the fluorescence per cell and objective evaluation of hybridization experiments. One of the probes developed was used to determine the population of Bifidobacterium spp. in human fecal samples. A comparison was made with results obtained by cultural methods for enumeration. Since both methods gave similar population estimates, it was concluded that all bifidobacteria in feces were culturable. However, since the total culturable counts were only a fraction of the total microscopic counts, the contribution of bifidobacteria to the total intestinal microflora was overestimated by almost 10-fold when cultural methods were used as the sole method for enumeration. PMID- 7487041 TI - Prevalence of Borrelia burgdorferi sensu lato-infected ticks on migrating birds. AB - The prevalence of Lyme disease Borrelia-infected ticks on migrating birds was studied in Scandinavia. A total of 22,998 birds were caught at eight different bird observatories and examined for ticks. Five different species of ticks were found infesting the birds. The dominant species, Ixodesricinus, constituted 98.3% of the ticks collected. The presence of spirochetes was determined by an immunofluorescence assay of tick larvae and DNA amplification by PCR on all ticks. To determine which Borrelia burgdorferi sensu lato species were present, a species classification was performed by DNA amplification with species-specific 16S rDNA primers and by DNA sequencing (rDNA is DNA coding for rRNA). Flagellin gene sequences of all species of B. burgdorferi sensu lato previously recorded in Europe were observed. Borrelia garinii was the most prevalent Lyme disease Borrelia species in ticks collected from birds arriving from the South or Southeast in the spring, whereas the distribution was more heterogeneous in ticks from birds migrating from the Southwest. These data support the notion that birds are partly responsible for the heterogeneous distribution of Lyme disease Borrelia spirochetes in Europe. PMID- 7487042 TI - Substrate specificities of bacterial polyhydroxyalkanoate depolymerases and lipases: bacterial lipases hydrolyze poly(omega-hydroxyalkanoates). AB - The substrate specificities of extracellular lipases purified from Bacillus subtilis, Pseudomonas aeruginosa, Pseudomonas alcaligenes, Pseudomonas fluorescens, and Burkholderia cepacia (former Pseudomonas cepacia) and of extracellular polyhydroxyalkanoate (PHA) depolymerases purified from Comamonas sp., Pseudomonas lemoignei, and P. fluorescens GK13, as well as that of an esterase purified from P. fluorescens GK 13, to various polyesters and to lipase substrates were analyzed. All lipases and the esterase of P. fluorescens GK13 but none of the PHA depolymerases tested hydrolyzed triolein, thereby confirming a functional difference between lipases and PHA depolymerases. However, most lipases were able to hydrolyze polyesters consisting of an omega-hydroxyalkanoic acid such as poly(6-hydroxyhedxanoate) or poly(4-hydroxybutyrate). The dimeric ester of hydroxyhexanoate was the main product of enzymatic hydrolysis of polycaprolactone by P. aeruginosa lipase. Polyesters containing side chains in the polymer backbone such as poly (3-hydroxybutyrate) and other poly(3 hydroxyalkanoates) were not or were only slightly hydrolyzed by the lipases tested. PMID- 7487043 TI - Characterization of streptomyces lydicus WYEC108 as a potential biocontrol agent against fungal root and seed rots. AB - The actinomycete Streptomyces lydicus WYEC108 showed strong in vitro antagonism against various fungal plant pathogens in plate assays by producing extracellular antifungal metabolites. When Pythium ultimum or Rhizoctonia solani was grown in liquid medium with S. lydicus WYEC108, inhibition of growth of the fungi was observed. When WYEC108 spores or mycelia were used to coat pea seeds, the seeds were protected from invasion by P. ultimum in an oospore-enriched soil. While 100% of uncoated control seeds were infected by P. ultimum within 48 h after planting, less than 40% of coated seeds were infected. When the coated seeds were planted in soil 24 h prior to introduction of the pathogen, 96 h later, less than 30% of the germinating seeds were infected. Plant growth chamber studies were also carried out to test for plant growth effects and for suppression by S. lydicus WYEC108 of Pythium seed rot and root rot. When WYEC108 was applied as a spore-peat moss-sand formulation (10(8) CFU/g) to P. ultimum-infested sterile or nonsterile soil planted with pea and cotton seeds, significant increases in average plant stand, plant length, and plant weight were observed in both cases compared with untreated control plants grown in similar soils. WYEC108 hyphae colonized and were able to migrate downward with the root as it elongated. Over a period of 30 days, the population of WYEC108 colonized emerging roots of germinating seeds and remained stable (10(5) CFU/g) in the rhizosphere, whereas the nonrhizosphere population of WYEC108 declined at least 100-fold (from 10(5) to 10(3) or fewer CFU/g). The stability of the WYEC108 population incubated at 25 degrees C in the formulation, in sterile soil, and in nonsterile soil was also evaluated. In all three environments, the population of WYEC108 maintained its size for 90 days or more. When pea, cotton, and sweet corn seeds were placed into sterile and nonsterile soils containing 10(6) or more CFU of WYEC108 per g, it colonized the emerging roots. After a 1-week growing period, WYEC108 populations of 10(5) CFU/g (wet weight) of root were found on pea roots in the amended sterile soil environment versus 10(4) CFU/g in amended nonsterile soil. To further study the in vitro interaction between the streptomycete and P. ultimum, mycelia of WYEC108 were mixed with oospores of P. ultimum in agar, which was then used as a film to coat slide coverslips.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 7487045 TI - Nucleotide sequence and replication properties of the Bacillus borstelensis cryptic plasmid pHT926. AB - The nucleotide sequence of pHT926, a cryptic plasmid found in Bacillus borstelensis HP926, was determined. pHT926 replicates by a rolling-circle mechanism and belongs to the pC194 plasmid family. The copy number of pHT926 was fourfold higher than that of pUB110 and very stably maintained in Bacillus choshinensis. PMID- 7487044 TI - Cloning and characterization of a gene encoding a secreted tripeptidyl aminopeptidase from Streptomyces lividans 66. AB - The gene encoding a tripeptidyl aminopeptidase (Tap) from Streptomyces lividans was cloned by using a simple agar plate activity assay. Overexpression of the cloned gene results in the production of a secreted protein which has an apparent subunit molecular weight of 55,000 and is responsible for the major amino terminal degradative activity in culture broths of S. lividans strains. A DNA sequence analysis revealed a potential protein-encoding region of the size expected to encode the observed protein, which contained a sequence that exhibited significant homology around a putative active site serine residue observed for lipases, esterases, and acyl transferases. Preceding the amino terminus of the secreted protein was a predicted signal peptide of 36 amino acids followed by a tripeptide, which could be autocatalytically removed from a secreted Tap precursor. The transcriptional start site for the gene was mapped by primer extension. Mutant strains of S. lividans lacking detectable Tap activity were able to grow and sporulate normally. Cross-species hybridization experiments showed that DNA homologs of the tap gene are present in most of the Streptomyces strains tested. PMID- 7487046 TI - Salmonella enteritidis phage type 4 isolates more tolerant of heat, acid, or hydrogen peroxide also survive longer on surfaces. AB - In a comparative study of different Salmonella enteritidis phage type 4 isolates we found that those isolates with enhanced heat tolerance also survived better than isolates that were heat sensitive either at pH 2.6, in 10 mM H2O2, or on surfaces. Culture to the stationary phase increased the heat tolerance of all isolates and the acid and H2O2 tolerance of heat-tolerant isolates. With heat sensitive isolates, however, extended culture had no impact on survival in H2O2 and only a marginal impact on acid tolerance. The growth phase had no appreciable impact on the surface survival of any of the isolates. PMID- 7487047 TI - Quantitative analysis of bacterial aerosols in two different dental clinic environments. AB - Microbial aerosols are generated during dental treatments and may represent an important source of infection. This study was designed to quantify bacterial air contamination during dental treatments in both a closed dental operatory and a multichair dental clinic. Air was sampled by using a slit type of biological air sampler. Following air sampling, blood-supplemented Trypticase soy agar plates were incubated at 37 degrees C under anaerobic conditions for 7 days. The maximum levels of air contamination in the closed dental operatory were observed while dental treatments were being performed (four trials; 216 +/- 75 CFU/m3 for ultrasonic scaling treatments and 75 +/- 22 CFU/m3 for operative treatments). At 2 h after completion of the treatments, the bacterial counts were about the same as the pretreatment levels (12 to 14 CFU/m3). In the second part of the study, a multichair dental clinic was divided into four areas, and air contamination was monitored at each site. Three sites were located in active dental treatment areas, whereas no dental treatments were performed within an 11-m radius of the fourth site. At 3 h after the beginning of dental treatments, the highest bacterial counts were obtained in the three active dental treatment areas (76 to 114 CFU/m3). However, there was noticeable contamination in the inactive dental treatment area (42 CFU/m3). Thus, bacterial aerosols were able to spread into areas where there was no dental activity. My data show that dental treatments significantly increased the levels of bacterial air contamination in both a closed dental operatory and a multichair dental clinic.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7487048 TI - Cloning, nucleotide sequencing, and expression of an opine dehydrogenase gene from Arthrobacter sp. strain 1C. AB - The gene coding for opine dehydrogenase from Arthrobacter sp. strain 1C was cloned onto plasmid pBluescript KS(-), and the nucleotide sequence of the 1,077 bp open reading frame consisting of 359 codons was identified as the odh gene. Transformed Escherichia coli cells overproduced NAD+-dependent opine dehydrogenase under control of the promoter of the lac gene on pBluescript KS(-). PMID- 7487049 TI - Characterization of plasmid-encoded citrate permease (citP) genes from Leuconostoc species reveals high sequence conservation with the Lactococcus lactis citP gene. AB - The citrate permease determinant (citP) in several Leuconostoc strains was demonstrated to be plasmid encoded by curing experiments and hybridization studies with a DNA fragment containing the citP gene from Lactococcus lactis subsp. lactis biovar diacetylactis NCDO176. Cloning and nucleotide sequence analysis of Leuconostoc lactis NZ6070 citP revealed almost complete identity to lactococcal citP. PMID- 7487052 TI - Degradation of p-xylene by a denitrifying enrichment culture. AB - Microbial cultures enriched from a diesel fuel-contaminated aquifer were able to grow on p-xylene under denitrifying conditions. The oxidation of p-xylene to CO2 was coupled to the reduction of NO3-. The enrichment cultures also grew on toluene and m-xylene, but they did not degrade benzene, ethylbenzene, and o xylene. PMID- 7487050 TI - Use of pulsed-field gel electrophoresis to link sporadic cases of invasive listeriosis with recalled chocolate milk. AB - Pulsed-field gel electrophoresis established the linkage between recalled chocolate milk and a multistate invasive listeriosis outbreak during a four product recall period. Listeria monocytogenes isolates from four hospitalized patients and an environmental dairy sample displayed AscI restriction endonuclease digestion profiles identical to that of the chocolate milk isolate. PMID- 7487051 TI - 3-Methylindole (skatole) and indole production by mixed populations of pig fecal bacteria. AB - Pig fecal slurries converted added L-tryptophan either to indole without detectable intermediates or to 3-methylindole (skatole) via indole-3-acetate. The initial rate of production of 3-methylindole was greatest at pH 6.5 and less at pH 5.0 and 8.0; the initial rates of indole production were similar at pH 6.5 and 8.0. More than 80% of the tryptophan added was converted to 3-methylindole at pH 5.0; at pH 8.0 85% was converted to indole. Both pathways had similar Km values for tryptophan and similar maximum rates. Indole-3-carbinol and indole-3 acetonitrile completely inhibited the production of 3-methylindole from indole-3 acetate but had no effect on the reactions involving L-tryptophan. PMID- 7487054 TI - A kinetic study of the hydrolysis of the N-tosylalanine ester of 3-hydroxy-5 phenylpyrrole and related compounds by human leukocyte elastase. AB - The tosylalanine ester of 3-hydroxy-5-phenylpyrrole (HOPPy) is localized on reagent strips (Ames LEUKOSTIX) and used diagnostically to test urine for the presence of human leukocyte elastase (HLE) as an indication of urinary tract infection. We have determined the kinetic constants for the HLE-catalyzed hydrolysis of this substrate and the related substrates Tos-Ala-ONp, Cbz-Ala OPPy, and Cbz-Ala-ONp, in solution at three different pH values. In the reagent strip matrix, diazo coupling of 4-diazo-3-hydroxy-1-napthylsulfonate (HONapN+2) with the enzymatic hydrolysis product HOPPy generates a purple color. We have also studied the kinetics of the reaction of HOPPy with HONapN+2 and other related diazonium salts such as 4-diazo-3-hydroxy-7-nitro-1-napthylsulfonate, 4 diazo-3-hydroxy-1,7-napthyldisulfonate, and 2-methoxy-4-(N-morpholinyl)benzene diazonium chloride. Tos-Ala-OPPy is the most reactive substrate among the compounds examined and kinetic studies indicate that deacylation is rate-limiting for HLE hydrolysis. The presence of decanol accelerates the enzymatic hydrolysis of Tos-Ala-OPPy with a kcat/KM = 10(7) M-1 s-1, which is close to the diffusion controlled limit. For the diazo coupling reaction, the rate is affected by the substituents on the naphthalene ring and by the buffer in which the reaction occurs. This research has elucidated some important mechanistic features for the reaction of these compounds and may lead to improved methods for the detection of leukocyte elastase. PMID- 7487053 TI - Divalent metal cation requirement and possible classification of cGMP-inhibited phosphodiesterase as a metallohydrolase. AB - cGMP-inhibited phosphodiesterase (cGI-PDE) has been found to require a divalent metal cation for cAMP hydrolysis. The cGI-PDE isolated from human platelets exhibited significantly higher enzymatic activity when incubated with Mn2+, Mg2+, and Co2+. The addition of Zn2+, Cd2+, Ca2+, K+, or Na+ to the enzyme did not enhance the activity and, when present in high concentration (> 1.0 microM), Zn2+ and Cd2+ inhibited the enzymatic activity of cGI-PDE. The inhibition by Zn2+ (and Cd2+) was partially prevented by preincubation of the enzyme with Mn2+. The enzyme was also inhibited by metal chelators EDTA and 1,10-phenanthroline and not by their non-metal-chelating analogs. The partial protection against chelation (and inhibition) was afforded by AMP (the product of cAMP hydrolysis). PMID- 7487056 TI - Engineering magnesium selectivity in the helix-loop-helix calcium-binding motif. AB - Engineering magnesium selectivity into the helix-loop-helix (hlh) cation binding site is relatively unstudied in the calmodulin superfamily of calcium-regulated proteins, which include parvalbumin, oncomodulin, troponin C, calbindin, and calmodulin. Studies using a 33-residue synthetic peptide model of the hlh cation binding motif have indicated that magnesium will induce structural change in those peptide motifs containing three or four acid residues in chelating positions with a single-acid-pair on the Z-axis. Decreasing the cation binding cavity size in Z-axis acid-paired motifs through replacement of chelating residues in the +Z or -X metal ion coordinating positions in the loop region by glutamic acid has been successful in decreasing the calcium ion affinity. The same changes did not create or enhance magnesium binding in the 33-residue model hlh cation binding motif. PMID- 7487057 TI - Kinetics and mechanisms of hypochlorous acid reactions. AB - Hypochlorous acid (HOCl) is a strong oxidant formed in neutrophils by the myeloperoxidase-catalyzed oxidation of chloride. Using stopped-flow with spectrophotometric detection, HOCl was found to react very rapidly with glutathione and ascorbate and less rapidly with taurine. No evidence could be found for the formation of reactive free-radical intermediates in these reactions, in support of an electrophilic mechanism. In contrast, the reaction with iron(II) aquo or citrate complexes (k approximately 10(4) dm3 mol-1 s-1 in acidic solution) yielded reactive intermediates distinguishable from hydroxyl radicals. The reaction between HOCl and ferrous ions, which is analogous to but faster than the Fenton reaction, is a potential source of free radicals in activated neutrophils. PMID- 7487055 TI - Analysis of melanoma cells stably transfected with beta 1,4GalNAc transferase (GM2/GD2 synthase) cDNA: relative glycosyltransferase levels play a dominant role in determining ganglioside expression. AB - Previous studies (Y. Nagata, S. Yamashiro, J. Yodoi, K. O. Lloyd, H. Shiku, and K. Furukawa (1992) J. Biol. Chem. 267, 12082-12089) had isolated a putative cDNA coding for beta 1,4 N-acetylgalactosaminyltransferase (GM2/GD2 synthase) and demonstrated the presence of GM2 and/or GD2 gangliosides in melanoma cell lines stably transfected with this gene. We have now measured the levels of glycosyltransferase activities and mRNA levels in five transfected cell lines in comparison with their parent cell lines (mouse melanoma B16 and human melanoma MeWo). Membrane preparations from the transfected cell lines demonstrated de novo synthesis of GM2 or GD2 in in vitro assays using GM3 or GD3, respectively, as ganglioside acceptors. The enzyme levels, however, varied considerably among the different transfectants, as did the mRNA levels for the beta 1,4 Gal-NAc transferase. The effect of the transfected gene on levels of preexisting glycosyltransferases involved in ganglioside biosynthesis was also measured and the ganglioside composition of the cell lines was determined. The level of beta 1,4 GalNAc-transferase expressed in the different cell lines was found to dramatically influence the overall ganglioside composition of the cell. In the transfected cell line with the highest levels of the transferase, for example, biosynthesis was almost completely redirected away from the b pathway to the a pathway with the resulting expression of only GM2. These data from this family of related cell lines clearly demonstrate the primary roles that relative glycosyltransferase levels play in determining the ganglioside composition of cells. PMID- 7487058 TI - Macrophages require both iron and copper to oxidize low-density lipoprotein in Hanks' balanced salt solution. AB - The oxidation of low-density lipoprotein (LDL) may be important in the pathogenesis of atherosclerosis. However, the interactions between cells and metals in promoting LDL oxidation are inadequately understood. A sensitive high performance liquid chromatography analysis of cholesterol, cholesteryl esters, and their oxidation products was used to identify and accurately measure LDL oxidation achieved in thiol-free Hanks' balanced salt solution (HBSS) at pH 7.4. Mouse peritoneal macrophages inhibited LDL oxidation when incubated in HBSS containing either 10 microM iron or 1 microM copper, but were markedly prooxidant in the presence of both metals. The prooxidant effect of macrophages in the presence of both iron and copper did not require the provision of added disulfides or thiols. Both Fe2+ and macrophages were demonstrated to independently reduce Cu2+ to Cu1+ in HBSS, indicating that the direct reduction of copper by cells or iron may underlie the observed promotion of LDL oxidation by macrophages in this system. We conclude that macrophages can either promote or inhibit metal-mediated LDL oxidation and that externally supplied thiols are not essential to the promotion of LDL oxidation by cells. The presence of both iron and copper may be particularly important for macrophages to promote LDL oxidation in vivo. PMID- 7487059 TI - Action of beta-carotene as an antioxidant against lipid peroxidation. AB - The action and activity of beta-carotene as a radical-scavenging antioxidant against lipid peroxidation have been studied. beta-Carotene suppressed the free radical-mediated oxidations of methyl linoleate in benzene solution and soybean phosphatidylcholine liposomal membranes in aqueous dispersions in a dose dependent manner, but the antioxidant activity of beta-carotene was much smaller than that of alpha-tocopherol. beta-Carotene was 32 times less reactive toward peroxyl radical than alpha-tocopherol and approximately as reactive as 2,6-di tert-butyl-4-methylphenol in benzene solution. Toward carbon-centered radical, beta-carotene was 11 times less reactive than alpha-tocopherol. When beta carotene and alpha-tocopherol were present together in homogeneous solution, alpha-tocopherol was consumed predominantly and beta-carotene was spared. When they were incorporated simultaneously into the same dimyristoyl phosphatidylcholine liposomal membranes and the radicals were formed in the aqueous phase, alpha-tocopherol was consumed faster than beta-carotene, but the sparing efficacy was much smaller than in homogeneous solution. On the contrary, beta-carotene was consumed faster than alpha-tocopherol when the radicals were generated within the lipophilic compartment of the membranes, implying that beta carotene is relatively more favorable than alpha-tocopherol for scavenging lipophilic radicals within the membranes. In contrast to an efficient synergistic inhibition by a combination of alpha-tocopherol and ascorbic acid, cooperative interaction between beta-carotene and ascorbic acid was not observed. beta Carotene underwent autooxidation to give polymeric products and the rate of consumption of beta-carotene increased with increasing concentrations of itself and oxygen and decreased with increasing concentration of lipids. The formation of polymeric products was confirmed by gel permeation chromatography. It was concluded that beta-carotene is less potent as an antioxidant than alpha tocopherol because beta-carotene is less reactive toward peroxyl radical than alpha-tocopherol and the stable beta-carotene radical reacts with oxygen to give beta-carotene peroxyl radical which is not stable but able to attack lipid to continue chain oxidation. The higher antioxidant activity of beta-carotene at lower oxygen pressure is attributed primarily to an unfavorable formation of beta carotene peroxyl radical at higher oxygen pressure rather than a higher reactivity of beta-carotene toward carbon-centered radical than peroxyl radical. PMID- 7487060 TI - Aminoacylpyrrolidine-2-nitriles: potent and stable inhibitors of dipeptidyl peptidase IV (CD 26). AB - Dipeptidyl-peptidase IV (EC 3.4.14.5) also known as CD26 is a membrane-bound serine peptidase which cleaves N-terminal dipeptides from a peptide chain containing a proline residue in the penultimate position. The enzyme is believed to play an important role in neuropeptide metabolism and T-cell activation. A series of aminoacylpyrrolidine-2-nitriles, in which the carboxyl group of proline is replaced by a nitrile group, was synthesized as inhibitors of dipeptidyl peptidase IV. All compounds were found to competitively inhibit a homogeneous preparation of the rat kidney enzyme with Ki values in the low to submicromolar range. The nitriles presumably react with the active-site serine to form an imidate adduct. The compounds were stable following incubation either for 20 h at 37 degrees C or 72 h at room temperature. They proved to be poor inhibitors of dipeptidyl-peptidase II and prolyl oligopeptidase. These studies demonstrate that the generally held concept that nitriles are poor inhibitors of serine proteinases needs to be reconsidered. Amino-acylpyrrolidine-2-nitriles by virtue of their ease of synthesis, stability, specificity, and inhibitory potency appear to be superior to other described dipeptidyl-peptidase IV inhibitors. PMID- 7487061 TI - Expression of spinach nitrite reductase in Escherichia coli: site-directed mutagenesis of predicted active site amino acids. AB - Spinach ferredoxin-nitrite reductase is a chloroplast enzyme that contains a coupled [Fe4S4]-siroheme-active site and catalyzes the six-electron reduction of nitrite to ammonia. An expression system which produced enzymatically active spinach nitrite reductase (NiR) in Escherichia coli was developed in order to study the structure-function relationships of the coupled active site using site directed mutagenesis. The spinach NiR cDNA, without the sequences encoding the chloroplast transit peptide, was expressed as a beta-galactosidase fusion containing five additional amino acids at the N-terminus. The expressed NiR in aerobic cultures was mostly insoluble and inactive. After optimizing growth conditions, active NiR represented 0.5-1.0% of the total protein. E. coli expressed NiR was purified approximately 200-fold to homogeneity as indicated by SDS-polyacrylamide gel electrophoresis. The expressed NiR enzyme was recognized by rabbit anti-spinach NiR antibody as visualized by Western blot analysis. The absorption spectrum of the E. coli-expressed NiR was identical to authentic spinach NiR with a Soret and alpha band at 386 and 573 nm, respectively, and a A278/A386 = 1.9. The addition of nitrite to the oxidized enzyme preparation produced the characteristic shifts in the spectrum. The specific activity for the methyl viologen-dependent reduction of nitrite of E. coli-expressed NiR was 100 U/mg and the Km determined for nitrite was 0.3 mM, which are in agreement with reported values for this enzyme. These results indicate that the E. coli expressed NiR is fully comparable to spinach NiR in purity, catalytic activity, and physical state.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7487062 TI - Purification and characterization of a fully active recombinant tobacco cytosolic NADP-dependent isocitrate dehydrogenase in Escherichia coli: evidence for a role for the N-terminal region in enzyme activity. AB - The recently isolated full-length NADP-dependent isocitrate dehydrogenase (ICDH) cDNA encoding the tobacco cytosolic isoenzyme has been cloned into the expression vector pET8c and used to transform Escherichia coli strain BL21 (DE3). The recombinant protein was purified to electrophoretic homogeneity and used to raise polyclonal antibodies. Its kinetic properties were found to be identical to those of the cytosolic ICDH isoenzyme purified from tobacco cell cultures. The recombinant and the endogenous bacterial ICDH could be easily distinguished by their different behaviors during anion-exchange column chromatography and immunological response. An incomplete ICDH-encoding cDNA clone, encoding a protein lacking the first 36 amino acids at the N-terminus, was cloned into the expression vector pKK233-2 and used to transform ICDH-lacking E. coli cells (strain 2004). The truncated, recombinant ICDH produced by the bacteria was found to be inactive. PMID- 7487063 TI - Dissociation and reassembly of the dihydrolipoyl transacetylase component of the bovine heart pyruvate dehydrogenase complex. AB - The catalytic activity and the state of aggregation of the dihydrolipoyl transacetylase-lipoamide dehydrogenase binding protein (E2-E3BP) subcomplex of the bovine heart pyruvate dehydrogenase multienzyme complex were investigated. Treatment of E2-E3BP with the chaotropic salts GndnCl or KSCN led to a rapid decrease in transacetylase activity which was accompanied by a loss of the native quaternary structure, as indicated by changes in the sedimentation properties of the E2-E3BP subcomplex. Reassembly or refolding of dissociated E2-E3BP was achieved for the GndnCl-treated subcomplex using a defined protocol. This reassembly procedure effectively excluded all E3BP from the reassembled oligomeric transacetylase. The reassembled oligomeric E2, free of E3BP, was unable to reconstitute the overall activity of the complex following incubation with pyruvate dehydrogenase (E1) and lipoamide dehydrogenase (E3). In binding studies using radiolabeled components it was demonstrated that the reassembled transacetylase, while retaining its capacity for reductive acetylation and its ability to bind E1, lost its ability to bind E3. The evidence presented in this study indicates that the strong association of E3BP with E2 facilitates the binding of E3, the lipoamide dehydrogenase component, and therefore may have an important role in the assembly and ultimately the catalytic activity of the pyruvate dehydrogenase multienzyme complex. PMID- 7487064 TI - The primary sequence of cytochrome P450tyr, the multifunctional N-hydroxylase catalyzing the conversion of L-tyrosine to p-hydroxyphenylacetaldehyde oxime in the biosynthesis of the cyanogenic glucoside dhurrin in Sorghum bicolor (L.) Moench. AB - The heme thiolate protein cytochrome P450tyr is a multifunctional N-hydroxylase converting L-tyrosine to p-hydroxyphenylacetaldehyde oxime in the biosynthesis of the cyanogenic glucoside dhurrin in Sorghum bicolor (Sibbesen et al. (1995) J. Biol. Chem. 270, 3506-3511). Using a polyclonal antibody toward cytochrome P450tyr and oligonucleotide probes designed on the basis of amino acid sequences of tryptic fragments, a full-length cDNA clone encoding cytochrome P450tyr has been isolated and sequenced. The open reading frame encodes a protein with a molecular mass of 61,887 Da. A comparison with the amino acid sequencing data demonstrates that the protein is not subjected to posttranslational modification at the N- and C-terminal ends except for the removal of the N-terminal methionine residue. Highest positional identity (30.8%) is found to the 3',5'-flavonoid hydroxylase of petunia (CYP75A1) and to a cytochrome P450 sequence from avocado of unknown function (CYP71A1). Consequently, cytochrome P450tyr is assigned as the first member of a new cytochrome P450 family denoted CYP79. The N-terminal region of cytochrome P450tyr contains the four domains characteristic for cytochrome P450 enzymes of the endoplasmic reticulum (ER) in animals. The amino acid sequence before the proline-rich domain is longer in cytochrome P450tyr and in four cytochrome P450s presently available from other monocotyledoneous plants compared to the sequences from dicotyledoneous plants but is concluded to contain a single transmembrane helix with the N-terminal located in the lumen of the ER and the bulk of the protein protruding into the cytoplasm. The heme-binding cysteine residue of cytochrome P450tyr is recognizable at position 493 but this region deviates from the consensus sequence by having an unusual alanine residue at position 495. The central region of helix I contains three residues, Ala-352, Asn-355, and Pro-356, deviating from the consensus sequence. CYP56 is the only other known cytochrome P450 using tyrosine as substrate and contains the same Asn Pro substitution in the consensus sequence of helix I indicating the importance of these residues in defining substrate specificity. The conserved threonine residue which normally helps to form the oxygen binding pocket is absent. The cytochrome P450tyr sequence represents the first amino acid sequence of a functionally characterized cytochrome P450 enzyme from a monocotyledoneous plant and the first sequence of a membrane-bound N-hydroxylase with high substrate specificity. Multifunctional N-hydroxylases of the cytochrome P450 type have not been previously demonstrated to catalyze biosynthetic pathways in living organisms. PMID- 7487065 TI - Candida albicans nucleoside-diphosphate kinase: purification and characterization. AB - Soluble nucleoside-diphosphate kinase (NDP kinase) from Candida albicans was purified to electrophoretic homogeneity and a partial sequence was determined. The enzyme was kinetically and physically characterized. Sodium dodecyl sulfate polyacrylamide gel electrophoresis of the purified enzyme revealed a single polypeptide of 17 kDa upon staining, by immunodetection with heterologous and homologous antibodies, and by autoradiography of the phosphorylated enzyme. Furthermore, isoelectric focusing of the purified native enzyme showed a single acidic band (pI 4.5). These results together with a native molecular mass of 98 kDa suggest a hexameric native enzyme composed of identical subunits. Like NDP kinases from other sources, the catalysis involves a phosphoenzyme intermediate that is rapidly formed upon incubation of the enzyme with ATP. The transfer of phosphate from phosphoprotein intermediate to nucleoside diphosphates is equally fast. Kinetic experiments indicated that GTP and ATP had the lowest Km compared to UTP, dTTP, and CTP. GDP acted as a preferred acceptor as assessed by kinetic measurements as well as by competition experiments. Experimental data are presented indicating the existence of a membrane-associated NDP kinase. Preliminary characterization of this enzyme suggests that cytosolic and membrane associated NDP kinases are similar proteins. PMID- 7487066 TI - Sites of electron transfer to membrane-bound copper and hydroperoxide-induced damage in the respiratory chain of Escherichia coli. AB - Previous studies in Escherichia coli as a model system for peroxide toxicity (L. Rodriguez-Montelongo, L. C. De la Cruz-Rodriguez, R. N. Farias, and E. M. Massa, 1993, Biochim. Biophys. Acta 1144, 77-84) have shown that electron flow through the respiratory chain supports a membrane-associated Cu(II)/Cu(I) redox cycle involved in irreversible impairment of the respiratory system by tert-butyl hydroperoxide (t-BOOH). In this paper, E. coli mutants deficient in specific respiratory chain components have been used to determine the sites of copper reduction and the targets inactivated by t-BOOH. Two sites of electron transfer to membrane-bound copper were identified: one in the region between NADH and ubiquinone supported by NADH as electron donor and another localized between ubiquinone and the cytochromes supported by electrons coming from NADH, succinate, or D-lactate. Electron flow through the former site in the presence of t-BOOH led to inactivation of NADH dehydrogenase II, whereas electron flow through the latter site in the presence of the hydroperoxide led to damage of ubiquinone. In agreement with the above in vitro results with isolated membranes, copper-dependent inactivation of NADH dehydrogenase and ubiquinone was demonstrated in E. coli cells exposed to t-BOOH. It is proposed that the t-BOOH induced damage is a consequence of t-butylalkoxy radical generation through a Fenton-type reaction mediated by redox cycling of membrane-bound copper at those two loci of the respiratory chain. PMID- 7487067 TI - Adenosine-5'-triphosphate-sulfurylase from Arabidopsis thaliana and Escherichia coli are functionally equivalent but structurally and kinetically divergent: nucleotide sequence of two adenosine-5'-triphosphate-sulfurylase cDNAs from Arabidopsis thaliana and analysis of a recombinant enzyme. AB - ATP-sulfurylase, the first enzyme of sulfate assimilation, catalyzes the formation of adenosine-5'-phosphosulfate from ATP and sulfate. Here we report that the higher plant, Arabidopsis thaliana, contains a three-member, expressed gene family encoding plastid localized forms of ATP sulfurylase. Three cDNAs from A. thaliana, designated APS1, APS2, and APS3, were isolated by their ability to functionally complement a met3 (ATP sulfurylase) mutant strain of Saccharomyces cerevisiae (yeast). The nucleotide sequence of APS1 was reported previously (1). APS2 and APS3, reported here, have 476- and 465-codon open-reading frames encoding 53.6- and 52.0-kDa polypeptides, respectively. The translation products of both clones are highly homologous to APS1 (66 and 86% identity, respectively) over their entire lengths, including amino terminal sequences resembling transit peptides for plastid localization. Both clones are less homologous to MET3 (25 and 30% identity, respectively). Genomic blot analysis of A. thaliana revealed only three genes with homology to the APS cDNAs and RNA blot analysis showed that APS1 is the most highly expressed member of this gene family. The APS polypeptides share homology with ATP-sulfurylases from fungi, a marine worm and a chemoautotrophic bacterium, but, not from Escherichia coli or Rhizobium meliloti. Analysis of recombinant APS3 showed that the protein is structurally and kinetically similar to fungal ATP-sulfurylase, but very different from the E. coli enzyme. The APS3 polypeptide is a homotetramer with specific activities (mumol primary product x mg protein-1 at pH 8.0, 25 degrees C) for 2.9 for APS synthesis, 30.1 for molybdolysis, and 48.7 for ATP synthesis. Despite the sequence, structural, and kinetic differences between higher plant and E. coli ATP-sulfurylases, APS2 and APS3 are able to functionally complement E. coli cysD and cysN (ATP-sulfurylase) mutant strains. PMID- 7487069 TI - Oxygen-dependent regulation of mitochondrial energy metabolism by nitric oxide. AB - To elucidate the role of nitric oxide (NO) in the regulation of mitochondrial function, the effect of NO on energy transfer reactions was examined under different oxygen tensions. Mitochondrial respiration was remarkably inhibited by NO resulting in the inhibition of ATP synthesis in a concentration-dependent manner. In the presence of succinate, respiration, respiratory control by ADP, and ATP synthesis recovered completely at certain times after adding NO. The inhibitory action of NO continued significantly longer under physiologically low oxygen concentrations (such as the cytosolic level) than at high concentrations. In the presence of various substrates, such as pyruvate malate, succinate, and ascorbate tetramethyl paraphenylenediamine, NO also inhibited the uncoupled respiration in an oxygen- and concentration-dependent manner. The inhibition of respiration by NO was stoichiometrically suppressed by oxyhemoglobin. When added to a mitochondrial suspension, NO rapidly disappeared from the medium particularly at high oxygen tension. However, the rate of NO disappearance was significantly lower under low oxygen tension. Thus, under cytosolic oxygen concentration, NO might play an important role in the regulation of mitochondrial energy metabolism. PMID- 7487068 TI - Topology of prostaglandin H synthase-1 in the endoplasmic reticulum membrane. AB - Prostaglandin H synthase-1 is an integral endoplasmic reticulum membrane protein which catalyzes a key control step in prostaglandin biosynthesis. The overall arrangement of the prostaglandin H synthase-1 polypeptide with respect to the endoplasmic reticulum membrane was examined in transiently transfected COS-1 cells, using immunofluorescence microscopy. A bacterial toxin, streptolysin-O, was used for selective plasma membrane permeabilization and a detergent, saponin, for general membrane permeabilization. Treated cells were probed with six antibodies specific for particular prostaglandin H synthase-1 peptide segments and one antibody specific for an inserted viral reporter epitope. Control experiments established that actin, a cytoplasmic marker, was accessible to fluorescein-labeled phalloidin after streptolysin-O treatment, whereas antibodies against protein disulfide isomerase, an endoplasmic reticulum lumenal marker, bound only after saponin treatment, Using this approach to investigate prostaglandin H synthase-1, it was found that streptolysin-O treatment was sufficient to obtain staining of intracellular membranes by antibodies specific for the endogenous C-terminal segment, for the viral reporter inserted at the C terminus, and for the protease-sensitive region near arg277. In contrast, saponin treatment was necessary for staining by antibodies specific for peptides spanning residues 51-66, 156-170, and 377-390. Antibodies targeted against residues 483 496 did not stain transfected cells even after saponin permeabilization, although they did bind to detergent-solubilized prostaglandin H synthase-1. These results indicate that the C-terminus and arg277 regions of the synthase can be exposed on the cytoplasmic side of the endoplasmic reticulum membrane, whereas regions near N-glycosylation sites are confined to the endoplasmic reticulum lumen and residues 483-496 are inaccessible from either side of the endoplasmic reticulum membrane. PMID- 7487070 TI - Chromate-induced chromium(V) formation in live mice and its control by cellular antioxidants: an L-band electron paramagnetic resonance study. AB - A recent note from our laboratory reported that L-band (1.2 GHz) electron paramagnetic resonance spectroscopy can be utilized in detecting the formation of Cr(V) intermediates from chromate-treated whole mice. Since Cr(V) is thought to be one of the key species in the mechanism of chromate's toxicity, we carried out additional measurements with improved sensitivity. The new spectra show partially resolved hyperfine structure from protons that suggests that the Cr(V) ion is ligated to NAD(P)H moieties via oxygens. Using laboratory-synthesized Cr(V) (K3CrO8) as a standard, the yield of Cr(V) formation was estimated to be 153 +/- 12 nmol after intravenous injection of 100 microliters of 100 mM sodium dichromate into mice. Pretreatment of the mice with ascorbic acid and glutathione significantly reduced the Cr(V) formation yield in a dose-related manner, while pretreatment with NADH had the opposite effect. Injection of ascorbic acid also had the effect of enhancing the rate of Cr(V) disappearance in vivo. By comparing these results with in vitro results utilizing L-band as well as X-band (9.6 GHz) measurements, we conclude that L-band spectroscopy can indeed be effectively utilized for following the metabolism of Cr(V) in live mice and that Cr(V) formation can be controlled by utilizing cellular antioxidants in vivo. PMID- 7487071 TI - Heterologous expression of selenium-dependent glutathione peroxidase affords cellular resistance to paraquat. AB - Transfection of murine NIH3T3 fibroblasts and human MCF7 breast carcinoma cells with a pSV2-derived eukaryotic expression vector for human cytosolic glutathione peroxidase resulted in clones with increased glutathione peroxidase activity. This heterologous expression indicates that murine cells recognize the human "selenocysteine insertion sequence" in the 3' untranslated region of the mRNA which facilitates insertion of selenocysteine directed by the opal codon. Though most clones from both cell lines eventually lost their enhanced glutathione peroxidase activity despite continuous selection on G418, some NIH3T3 clones retained enhanced enzyme activity without continuous G418 exposure. Transfection of MCF7 cells with an Epstein-Barr virus (EBV)-derived episomally replicating expression vector carrying the glutathione peroxidase gene also revealed increased glutathione peroxidase activity. These MCF7 cells, however, all required exposure to G418 to maintain enhanced glutathione peroxidase activity. Detailed biochemical analysis of a stably expressing NIH3T3 clone and MCF7 expressing cells revealed no alterations in activities of copper-zinc superoxide dismutase, manganese superoxide dismutase, catalase, phospholipid-glutathione peroxidase, glutathione reductase, glutathione transferase, or NADPH-P450 reductase. Both pSV2- and EBV-derived glutathione peroxidase-expressing clones exhibited enhanced resistance to paraquat as well as to peroxides. PMID- 7487073 TI - Identification of protein-receptor components required for the import of prealdehyde dehydrogenase into rat liver mitochondria. AB - Mitochondrial aldehyde dehydrogenase is synthesized as a high-molecular-weight precursor in cytosol and transported into mitochondrial matrix space where it is processed to the mature enzyme. To identify components of the transport machinery on liver mitochondria, anti-idiotypic antibodies against the rabbit anti prealdehyde dehydrogenase signal peptide antibodies were produced in chicken eggs and rabbit. Both anti-idiotypic antibodies inhibited the import of prealdehyde dehydrogenase (pALDH) into isolated rat liver mitochondria. The rabbit anti idiotypic antibody could recognize by Western blotting five mitochondrial membrane proteins with apparent molecular weights of 66, 60, 42, 34, and 29 kDa. The anti-idiotypic antibodies were cross-linked to mitochondrial membrane proteins using sulfosuccinimidyl 2-(p-azidosalicylamido)ethyl-1,3' dithiopropionate which is an iodinatable, heterofunctional, and photoreactive cross-linker. Mitochondrial proteins with apparent molecular weights of 66, 60, and 42 kDa were identified using the chicken antibody. The 66- and 34-kDa proteins were cross-linked to the rabbit antibody as the major components and the 42-kDa protein as a minor one. Antibodies against the 60- and 42-kDa proteins, as well as Fab fragments, inhibited the import of pALDH, suggesting that these proteins are receptor/translocator components for pALDH import. PMID- 7487074 TI - Induction of Ya1 subunit of rat hepatic glutathione S-transferases by exercise induced oxidative stress. AB - Adult female rats subjected to exercise training in the form of swimming daily (for 20 min initially with gradual increase up to 90 min within a week) for 130 days showed a significant elevation of liver glutathione S-transferase (GST) activity when compared to controls. Electrophoresis and Western blot analysis of cytosolic as well as affinity-purified hepatic glutathione S-transferases revealed the induction of Ya-sized subunit in exercise-trained animals. Reverse phase HPLC analysis of affinity-purified GSTs further revealed that Ya-sized subunits in control animals consisted of predominantly Ya2, whereas the trained animals displayed a dramatic 4.3-fold increase in Ya1. The GSTs of exercise trained animals showed increased peroxidase activity when compared to corresponding controls, which is consistent with the changes in sub-unit composition. This could be a response to the physiological oxidative stress induced by physical exercise. PMID- 7487072 TI - Expression, isolation, and characterization of the hepatitis C virus ATPase/RNA helicase. AB - The genome of the hepatitis C virus directs the synthesis of a single polyprotein, which is proteolytically cleaved into at least nine functional proteins. The amino-terminal portion of the polyprotein forms the structural proteins, while the carboxy-terminal region constitutes a variety of viral enzymes. The nonstructural 3 (NS3) protein, consisting of amino acids 1027-1657 of the polyprotein, is believed to be a multifunctional protein with an amino terminal serine protease domain, which is involved in polyprotein processing, and a carboxy-terminal ATPase/RNA helicase domain, presumably involved in viral replication. We have assembled an expression vector which directs the synthesis of residues 1207-1612 of the polyprotein with an amino-terminal polyhistidine purification tag. This portion of the NS3 protein contains the putative ATPase/helicase domain. The protein has been purified to yield 30-50 mg of enzymatically active protein per liter of culture. The purified NS3 protein has both NTPase and RNA helicase activities. ATP is the preferred substrate for the NTPase; GTP is also utilized; however, UTP is a very poor substrate and CTP is not utilized. The RNA helicase activity is dependent on ATP and divalent cation. Either manganese or magnesium can serve as the divalent cation. PMID- 7487075 TI - Generation of 5,5-dimethyl-1-pyrroline N-oxide hydroxyl and scavenger radical adducts from copper/H2O2 mixtures: effects of metal ion chelation and the search for high-valent metal-oxygen intermediates. AB - A metal-catalyzed nucleophilic addition mechanism for the formation of radical adducts of the spin trap 5,5-dimethyl-1-pyrroline N-oxide (DMPO) has been described recently (K. Makino, T. Hagiwara, A. Hagi, M. Nishi, and A. Murakami, 1990, Biochem. Biophys. Res. Commun. 172, 1073-1080; P. M. Hanna, W. Chamulitrat, and R. P. Mason, 1992, Arch. Biochem. Biophys. 296, 640-644). In the present investigation, we have demonstrated that the recently reported inhibition of copper-dependent hydroxyl radical formation by the complexing agent 1,10 phenanthroline (OP), which appears to contradict the well-known chemical nuclease properties of CuI(OP)2, is an artifact resulting from an inhibition of the nucleophilic addition of water to DMPO by OP (A. C. Mello-Filho and R. Meneghini, 1991, Mutat. Res. 251, 109-113). Copper bound to OP was found to be a good catalyst of hydroxyl radical formation: the CuII(OP)2 complex can be reduced by H2O2 and the CuI(OP)2 generated reacts with the peroxide to form .OH. In contrast, no evidence could be obtained for oxidant formation from the CuII(aq)/H2O2 reaction system, despite the detection of a prominent signal from the DMPO hydroxyl radical adduct (DMPO/.OH) (the formation of which was due solely to the nucleophilic addition of water to DMPO). The failure to generate an oxidant in this reaction mixture was attributed to the failure of hydrogen peroxide to reduce CuII(aq), as hydroxyl radical formation did occur when CuI(aq) was added directly to H2O2. However, in order to account for the high concentration of alpha-hydroxyethanol radicals detected when ethanol was included in the CuI(aq)/H2O2 reaction, the possibility that an oxidant in addition to .OH (e.g., CuO+) is generated is discussed. PMID- 7487076 TI - Differential activity of trans-acting hammerhead ribozymes targeted to beta amyloid peptide precursor mRNA by altering the symmetry of helices I and III. AB - In order to determine whether distributing the pairing bases in helices I and III of a hammerhead ribozyme asymmetrically would enhance the cleavage of trans acting ribozymes designed to degrade cellular mRNAs, we measured the cleavage properties of symmetric and asymmetric ribozymes targeted to the amyloid peptide precursor (beta APP) mRNA. Five ribozymes were formed from three beta APP synthetic mRNA analogs and two ribozyme RNA core sequences. Symmetric ribozymes beta 133/Rz133 and beta 125/Rz133 contained 8 bp in helix I and 7 bp in helix III. Asymmetric ribozyme beta 125/Rz125 had 13 bp in helix I and 4 bp in helix III, asymmetric ribozyme beta 123/Rz125 had 11 bp in helix I and 4 bp in helix III, while asymmetric ribozyme beta 133/Rz125 contained 8 bp in helix I and 4 bp in helix III. The ability of each ribozyme to cleave its substrate RNA was first assessed under single-turnover conditions at 37 degrees C. These studies revealed that only symmetric ribozyme, beta 133/Rz133, and asymmetric ribozyme beta 123/Rz125 effectively cleaved their substrates. Further studies using a 80 degrees C, 1-min-->37 degrees C, 1-min temperature cycling paradigm were performed to increase the cleavage efficiency of the ribozymes. Under these conditions ribozymes beta 133/Rz133, beta 125/Rz125, and beta 123/Rz125 were kinetically well behaved. Therefore, the fact that the symmetric ribozyme beta 133/Rz133 was more active than its asymmetric counterparts indicates that symmetrically distributing the pairing bases in helices I and III around this cleavage site is preferred. PMID- 7487077 TI - Recombinant human dihydroorotate dehydrogenase: expression, purification, and characterization of a catalytically functional truncated enzyme. AB - An N-terminally truncated cDNA for human dihydroorotate dehydrogenase (DHODase) was placed under the control of the inducible T7 lac promoter in a pyrimidine auxotrophic strain of Escherichia coli lacking the endogenous enzyme. Induction of gene expression rescued growth in media lacking exogenous pyrimidines. The recombinant enzyme was purified to homogeneity from detergent extracts of bacterial membranes by two chromatographic steps. The purity of the resulting enzyme was judged to be > 95% based on SDS-PAGE with Coomassie staining. The enzyme displays an apparent molecular weight of ca. 40 kDa on SDS-PAGE and ca. 120 kDa on native size-exclusion chromatography, suggesting that the native enzyme is multimeric. Recombinant DHODase displayed a specific activity and Km for dihydroorotate that were similar to those for the enzymes from bovine and human liver tissue. The pH dependence of the activity of the recombinant enzyme was likewise similar to that of the enzyme from human liver and may indicate the involvement of a critical histidine residue in catalytic turnover; only eight histidine residues remain in the truncated version of DHODase used here. The catalytic activity of the recombinant enzyme is inhibited in a dose-dependent fashion by the histidine-selective modifying agent diethylpyrocarbonate. These results further suggest a potential role for histidine in enzyme turnover. Brequinar sodium, an experimental drug which has been shown to be a nanomolar noncompetitive inhibitor of mammalian DHODases, inhibited the activity of the purified recombinant enzyme with a Ki value similar to that for enzyme derived from human liver tissue. The recombinant DHODase thus displays enzymatic behavior similar to the 50-kDa full-length human liver enzyme, illustrating that the catalytically essential structural features of the enzyme, as well as the site of Brequinar binding, are contained within the 40-kDa truncated version of the enzyme that was expressed here. PMID- 7487079 TI - Effect of linoleic acid hydroperoxide on endothelial cell calcium homeostasis and phospholipid hydrolysis. AB - The relationship between intracellular free calcium ion concentrations ([Ca2+]i) and release of arachidonic acid from membrane phospholipids following peroxidation was examined in rabbit aortic endothelial cells treated with linoleic acid hydroperoxide (LOOH). LOOH (0.1-0.4) mumol/10(6) cells) caused a rapid and dose-dependent transient increase in [Ca2+]i in the presence of extracellular Ca2+ that remained elevated over baseline for 15 to 30 s. In the absence of extracellular Ca2+, LOOH also evoked a transient increase in [Ca2+]i of lesser magnitude which immediately returned to basal (or below basal) levels. In this regard, the rise in intracellular Ca2+ after LOOH or vasopressin (AVP) treatments involved, at least in part, related intracellular pools that in each case was followed by influx of extracellular Ca2+. The intracellular membrane sources known to be affected by vasopressin were not directly involved. Most notably, the LOOH evoked rise in [Ca2+]i was not associated with release of IP3, suggesting that the source of intracellular Ca2+ is not IP3-sensitive pools. However, pretreatment with LOOH strongly inhibited the rise in [Ca2+]i upon subsequent addition of AVP or LOOH and the extent of such inhibition was dependent on the availability of free intracellular Ca2+ and presence of extracellular Ca2+. These findings suggest that reuptake of Ca2+ into intracellular membrane pools is reduced in the presence of LOOH and/or the availability of Ca2+ from agonist-sensitive sites is inhibited by LOOH. An increase in free 20:4 levels was found after LOOH treatment that was only partly prevented using intracellular Ca2+ chelators which maintained [Ca2+]i at basal levels after LOOH treatment. These findings suggest that LOOH induction of phospholipid hydrolysis proceeds following small transients in [Ca2+]i that are considerably less than that evoked by agents such as AVP, approximating basal Ca2+ concentrations. Inhibition of LOOH-induced lipid peroxidation by vitamin E also prevented the rise in [Ca2+]i and 20:4 release indicating that phospholipid hydrolysis is dependent, at least in part, on membrane lipid peroxidation. Inhibition of protein kinase C (PKC) completely blocked LOOH-induced release of 20:4 but had little effect on the LOOH-induced rise in [Ca2+]i, suggesting an indirect relationship between LOOH-induced membrane Ca2+ signalling events, with intervention via PKC-mediated induction of phospholipid hydrolysis. A rapid and progressive translocation of PKC to the membrane fraction was evident after LOOH addition over the time course corresponding to the maximal release of 20:4 which was also inhibited by vitamin E.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 7487078 TI - A universal approach to the expression of human and rabbit cytochrome P450s of the 2C subfamily in Escherichia coli. AB - Human cytochrome P450s 2C8, 2C9, 2C18, and 2C19 and rabbit cytochrome P450s 2C1, 2C2, 2C4, 2C5, and 2C16 were expressed from their respective cDNAs in Escherichia coli as chimeric enzymes in which a portion of the N-terminal membrane anchor sequence was replaced with a modified sequence derived from P450 17A. For 2C1 and 2C2 removal of the extraneous 3'-untranslated sequence allowed the successful expression of constructs that were unproductive in its presence. The levels of expression varied from 180 to 1500 nmol/liter of culture and the addition of delta-aminolevulinic acid to the culture media increased the amount of spectrally detectable P450 for several of these enzymes 2- to 10-fold. The catalytic properties of the modified human 2C P450s expressed in E. coli were concordant with previously published data for several marker substrates including (S) mephenytoin for P450 2C19, tolbutamide and tetrahydrocannabinol (THC) for P450 2C9, and taxol for P450 2C8. Interestingly, P450 2C19 catalyzed the 21 hydroxylation of progesterone and, to a lesser extent, catalyzed the formation of 16 alpha-hydroxyprogesterone. The rabbit enzyme P450 2C16 catalyzed the formation of 17 alpha- and 16 alpha-hydroxyprogesterone in addition to 21-hydroxylation. P450 2C19 also catalyzed the methylhydroxylation of tolbutamide and the 7 hydroxylation of THC at rates that were similar to or greater than that of P450 2C9. This work has identified important factors required for the high-level expression of 2C subfamily P450s in E. coli. The availability of these enzymes will facilitate detailed kinetic measurements for known and yet to be identified substrates. PMID- 7487080 TI - Reductant-independent ATP hydrolysis catalyzed by homologous nitrogenase proteins from Azotobacter vinelandii and heterologous crosses with Clostridium pasteuranium. AB - Reductant-independent ATPase activity was initiated and studied for Azotobacter vinelandii and Clostridium pasteuranium nitrogenase proteins (Av1, Cp1 and Av2, Cp2, 1 designating the iron molybdenum protein and 2 the iron protein) and their heterologous crosses by two methods: (1) allowing dithionite to be depleted from a normal assay in the presence of substrate levels of MgATP and (2) using reduced but reductant-free nitrogenase proteins in the presence of substrate levels of MgATP. In both cases, at a 1:1 protein ratio, MgATP is converted initially to MgADP with a specific activity of 400-500 nmol MgATP hydrolyzed/min.mg Av1, but in slower steps the MgADP is converted to AMP and, after 12 h, AMP is ultimately converted to adenosine. This reactivity requires the presence of both proteins, increases with increasing Av2/Av1 ratio, and is not a result of unique redox states of either protein. For Av1-Av2, ATP hydrolysis in the absence of Mg2+ occurred at nearly the same rate as reductant-dependent MgATP hydrolysis. Reductant-independent ATPase activity also occurred for the Av1-Cp2 and Cp1-Av2 heterologous crosses and was 2-fold and 18-fold slower than the Av1-Av2 or Cp1 Cp2 combinations. In both cases further hydrolysis of MgADP to AMP and AMP to adenosine occurred. A unique nucleotide hydrolysis system is apparently operating in the complex formed between the two nitrogenase proteins in the absence of reductant. The relationship between the reductant-independent and reductant dependent activities of nitrogenase catalysis is explored. PMID- 7487082 TI - Purification of the adenomatous polyposis coli (APC) gene product. AB - Mutation of the adenomatous polyposis coli (APC) gene is responsible for familial adenomatous polyposis and is an etiologic factor for digestive tract malignancies. Although the APC gene product (APC) is believed to play a role in growth suppression of colonocytes, the underlying mechanism is not clear. However, recent evidence does suggest that APC is a microtubule-associated protein (MAP), and like other MAPs, it can be phosphorylated, as we have shown. To facilitate studies of APC function, we purified the APC protein. To purify the full-length APC protein, HCT116 human colon cancer cells were lysed and the particulate fraction from the lysate was extracted with ammonium sulfate followed by Sepharose 4B and DEAE-Sephacel column fractionation and then by sucrose zonal density gradient centrifugation. The final purified APC fraction was determined to be about 1000-fold enriched in APC. The availability of purified APC will be valuable in investigating possible growth-suppressing mechanisms of APC including specific sites of APC phosphorylation and APC's interaction with other cellular proteins. PMID- 7487081 TI - Calmodulin-dependent cyclic AMP phosphodiesterase in liver plasma membranes: stimulated by insulin. AB - In vivo insulin consistently stimulates the plasma membrane, high-affinity (low Km) cyclic AMP phosphodiesterase (PDE) from diabetic rat liver or adipose tissue. In vitro stimulation of membrane PDE by insulin has been reported to be inconsistent. Also, the involvement of calmodulin (CaM) in insulin stimulation of PDE has been controversial. In this report, conditions for the isolation of rat liver plasma membranes containing PDE that is sensitive to in vitro insulin stimulation and the involvement of CaM in insulin stimulation of PDE were investigated. In vitro insulin raised the Vmax of the enzyme without altering its apparent Km and was dose dependent. Insulin stimulation was lost after freezing, sonication, solubilization with detergents, or storage of the membranes at 4 degrees C for 4 h after isolation. Insulin stimulation was completely blocked by the CaM antagonist compound 48/80, EGTA, or trifluoperazine. Two isoforms of membrane-bound PDE were separated by ion-exchange chromatography following solubilization of the plasma membranes. The activities of both isoforms were stimulated by exogenous CaM. Plasma membrane PDE eluted after the application of exogenous CaM plus Ca2+. The data support the concept of a critical involvement of CaM in insulin activation of liver membrane PDE. PMID- 7487084 TI - Induction of highly proliferative phenotypes in cultured glomerular mesangial cells by benzo[a]pyrene alone or in combination with methoxamine. AB - Recent studies have suggested that aromatic hydrocarbons can initiate glomerular mesangial cell (GMC) injury and contribute to the onset of renal disease. The present studies were conducted to assess the impact of benzo[a]pyrene (BaP), a ubiquitous polycyclic aromatic hydrocarbon, on the proliferation of GMCs. Challenge of cultured GMCs with BaP (0.3-30 microM) for 24 h was associated with concentration-dependent decreases in DNA synthesis, a response mediated by selective interference with early G1 cell cycle progression. One cycle of sequential treatment with 3 microM BaP for 24 h followed by challenge with 10 microM methoxamine (MeoA), a growth-promoting alpha 1-adrenergic agonist, for an additional 24 h attenuated the inhibitory response elicited by BaP alone. Following three rounds of sequential treatment with BaP and MeoA, GMCs exposed to BaP alone or BaP/MeoA exhibited enhanced proliferation rates relative to controls. BaP/MeoA cells acquired the greatest proliferative enhancement and exhibited unregulated c-jun and c-fos gene expression under growth-arrested and serum-stimulated conditions. Marked increases in specific AP-1 binding to a synthetic oligonucleotide were observed upon serum stimulation of quiescent cultures of BaP/MeoA cells relative to controls or any of the other treatment groups. These data demonstrate that sequential treatment with BaP in combination with MeoA is associated with induction of highly proliferative phenotypes in GMCs characterized by differential expression of growth-related protooncogenes. PMID- 7487083 TI - A tyrphostin-derived inhibitor of protein tyrosine kinases: isolation and characterization. AB - We recently reported that tyrphostin 23 (3,4-dihydroxybenzylidene malononitrile) is unstable in solution and that some of the degradation products are better inhibitors of the tyrosine kinase activity of Src and the EGF-receptor kinase than the parent compound itself (Ramdas et al., Cancer Res. 54, 867-868, 1994). In this study, the tyrphostin 23-derived compound designated P3, which is a more stable and potent protein tyrosine kinase inhibitor, was isolated. P3 was purified from oxidized tyrphostin 23 by solvent extraction, silica-gel flash chromatography, and reverse-phase high-pressure liquid chromatography. The physical characteristics of the isolated compound were determined and its chemical structure elucidated by 1H and 13C NMR spectroscopy. The proposed structure of this new inhibitor is that of a tyrphostin 23 dimer joined at the benzylidene carbon. P3 was evaluated in vitro as an inhibitor of four different protein tyrosine kinases (Src, Csk, EGF-receptor, and FGF-receptor) and two protein serine kinases (PK-A and PK-C). This compound exhibited the most inhibitory activity against Src with a Ki value of 6 microM and was less inhibitory toward the other protein kinases with Ki values ranging from 35 to 300 microM. P3 did not inhibit other nucleotide-utilizing enzymes such as lactate dehydrogenase and hexokinase. The growth and colony formation of HT-29 colon adenocarcinoma cells that contain activated Src was inhibited by P3 with an IC50 value of approximately 10 microM. PMID- 7487085 TI - Influence of calcium on arachidonic acid mobilization by murine resident peritoneal macrophages. AB - Stimulation of resident peritoneal macrophages resulted in release of arachidonic acid (AA) from phospholipids. This AA release is believed to occur as a result of the activation of phospholipases, usually by a phospholipase A2 (PLA2). The purpose of this study was to elucidate the role of cytosolic calcium ion concentration ([Ca2+]i) in the modulation of [3H]AA mobilization in peritoneal macrophages. [3H]AA release induced by ionophore A23187, opsonized zymosan, or 4 beta-phorbol 12-myristate acetate (PMA) occurred in the absence of extracellular calcium. Studies in fura-2/AM-loaded cells showed that zymosan and PMA did not increase [Ca2+]i significantly, whereas A23187 induced PLA2 activity translocation up to plasmatic membrane. Thapsigargin, an inhibitor of endomembrane Ca(2+)-ATPase, induced a rise in [Ca2+]i when cells were incubated in a Ca2+ medium. However, thapsigargin was not an effective stimulator of the translocation of PLA2 activity and [3H]AA release. These data indicate that changes in [Ca2+]i were not sufficient to elicit [3H]AA mobilization; this process seems tightly modulated by phosphorylation-dependent mechanisms in the presence of low [Ca2+]i. PMID- 7487086 TI - Chronic vitamin A intake affects the expression of mRNA for apolipoprotein A-I, but not for nuclear retinoid receptors, in liver of young and aging Lewis rats. AB - We have investigated the effects of chronic retinoid status and age on the steady state levels in rat liver of the mRNAs for the retinoid-responsive gene apolipoprotein A-I (apo A-I), and for three nuclear retinoid receptors, RAR alpha, RAR-beta, and RXR-alpha. Weanling male Lewis rats (five per age-diet group) were fed a semi-synthetic diet with vitamin A at marginal, control, or supplemented levels until they were 2 or 8 months of age. The relative abundance of apo A-I mRNA (compared to beta-actin which did not differ among treatments) was about 50% lower in aging versus young rats, regardless of their vitamin A status. For both ages, apo A-I mRNA was approximately 40-50% higher (P < 0.05) for the vitamin A-marginal groups and 20-30% lower (P > 0.05) for the vitamin A supplemented groups than the controls. Neither chronic diet nor age affected the levels of mRNA for RAR-alpha or RXR-alpha, two major retinoid receptors in liver, or for RAR-beta, a receptor known to be autoregulated by RA. Although vitamin A did not have a significant effect on the mRNA levels of the receptors examined in this study, they are still likely to be involved in regulating apo A-I gene expression because in vitro studies have shown that both RAR-alpha and RXR-alpha bind to the regulatory region of the apo A-I gene. PMID- 7487087 TI - Roles of histidine-194, aspartate-163, and a glycine-rich sequence of NAD(P)H:quinone oxidoreductase in the interaction with nicotinamide coenzymes. AB - NAD(P)H:(quinone-acceptor) oxidoreductase (NQOR, EC 1.6.99.2), an enzyme catalyzing the obligatory two electron reduction of quinones, can utilize both NADH and NADPH as electron donors at similar efficiencies. Based on site-directed mutagenesis studies, we previously suggested that the glycine-rich region of rat liver NQOR is important for the binding of NAD(P)H (Ma et al., J. Biol. Chem. 267, 22298-22304, 1992). However, the mode of interactions between the active site and NADH or NADPH is not clearly known. In this study, we conducted site directed mutagenesis experiments and identified H194 and D163 of NQOR as key residues affecting the Km of NADPH. Steady-state kinetic analysis for the reduction of dichloroindophenol (DCIP) showed that Km(NADPH) values of purified mutant proteins H194D, H194A, and D163V were 288-, 14-, and 96-fold higher, respectively, than that of NQOR; but the Km(NADH) values were only slightly higher. The kcat(NADPH) values were almost the same as that of NQOR in the reduction of DCIP at the respective pH optima which were affected by the mutations. The kcat(NADH) values of these mutant enzymes were 30 to 60% that of NQOR. In the reduction of menadione, the mutations also caused much larger increases in km(NADPH) than Km(NADH). The results suggest that H194 and D163 are important for the interaction with the 2'-phosphate group of NADPH. NAD(P)H analogues, N-methyldihydronicotinamide and dihydronicotinamide mononucleotide, can also serve as electron donors for NQOR, but the Km values were 4.5- and 495 fold higher, respectively, than that with NADH. Mutations at H194 and D163 and at the glycine-rich region of NQOR, which increased Km(NADH) and Km(NADPH), did not substantially affect the Km values of these two analogues. This result is consistent with the suggested roles of these amino acid residues in the interaction with nicotinamide coenzymes. Based on these results, a model of the NAD(P)H binding site is proposed showing the interaction of the pyrophosphate group with the glycine-rich region and the interaction of 2'-phosphate group with H194 and D163. PMID- 7487088 TI - Terminal enzymes of heme biosynthesis in the plant plasma membrane. AB - Purified plasma membrane fractions from barley leaf exhibited activities of ferrochelatase and iron reductase, two of the terminal enzymes in heme biosynthesis. These activities were also present in purified barley leaf plastid and mitochondrial fractions. The plasma membrane fractions were shown to be free from contamination with plastid and mitochondrial membrane markers. Previous studies had demonstrated that barley plasma membranes exhibited activity for converting protoporphyrinogen to protoporphyrin, another late step in heme synthesis. The presence of the terminal steps of heme synthesis in the plant plasma membrane is compatible with the hypothesis that late heme precursors such as protoporphyrin or protoporphyrinogen synthesized in the chloroplast can be exported and converted to heme within the plasma membrane for subsequent incorporation into plasma membrane hemoproteins. PMID- 7487090 TI - The expression of an intestinal form of glutathione peroxidase (GSHPx-GI) in rat intestinal epithelium. AB - We have previously identified and characterized GSHPx-GI, which is a cellular selenium-dependent glutathione peroxidase (GSHPx) distinct from the classic GSHPx 1 and phospholipid hydroperoxide glutathione peroxidase (PHGPX). We have determined the level of GSHPx-GI mRNA expression in the rat gastrointestinal tract from esophagus to colon. Although GSHPx-GI mRNA is readily detectable throughout the GI tract, the highest level is detected in the ileum and cecum. We have also determined the levels of GSHPx-GI mRNA expression and several antioxidant enzyme activities along the villus-to-crypt axis in the rat small intestine by cell fractionation. GSHPx-GI mRNA is present at a similar level in all of the epithelial fractions, whereas GSHPx-1 mRNA is detectable only in the remnant. This suggests that GSHPx-GI is the major cellular tetrameric GSHPx expressed in intestinal epithelium, and the expression of GSHPx-GI in the GI tract is not likely regulated differentially through maturation of epithelial cells. In terms of enzymatic activity, although we detected lower glutathione S transferase activity in the crypt epithelium, there was a marginal increase of PHGPX activity, a twofold increase of GSHPx activity, and a three- to fivefold increase of catalase activity in the crypt relative to the distal villus. Thus, the crypt epithelial cells may be better protected from peroxidative damage. PMID- 7487089 TI - Differences in the interaction of heparin with arginine and lysine and the importance of these basic amino acids in the binding of heparin to acidic fibroblast growth factor. AB - Although the interaction of proteins with glycosaminoglycans (GAGs) such as heparin are of great importance, the general structural requirements for protein- or peptide-GAG interaction have not been well characterized. Electrostatic interactions between sulfate and carboxylate groups on the GAG and basic residues in the protein or peptide dominate the interaction, but the thermodynamics of these electrostatic interactions have not been studied. Arginine residues occur frequently in the known heparin binding sites of proteins. Arginine is also more common than lysine in randomly synthesized 7-mer peptides that bind to immobilized heparin and heparan sulfate. We have used heparin affinity chromatography, equilibrium dialysis, and isothermal titration calorimetry techniques to further investigate these interactions. A 7-mer of arginine eluted from a heparin-affinity column at 0.82 M NaCl, whereas the analogous 7-mer of lysine eluted at 0.64 M. Similarly, the putative heparin binding site peptide (amino acid residues 110-130) from acidic fibroblast growth factor, which contained four lysine and two arginine residues, eluted at 0.50 M, whereas the analogous peptide with six lysine residues eluted at 0.41 M and one with six arginine residues eluted at 0.54 M. At 25 degrees C in 10 mM sodium phosphate, pH 7.4, carboxy and amino termini blocked arginine (blocked arginine) bound to heparin twice as tightly as blocked lysine as measured by equilibrium dialysis Similarly, at 30 degrees C in 10 mM sodium phosphate, pH 7.4, and in water, blocked arginine bound 2.5 times more tightly to anions in heparin than blocked lysine. Using titration calorimetry, the enthalpy of blocked arginine and lysine binding to heparin was 1.14 +/- 0.24 and 0.45 +/- 0.35 kJ/mol, respectively, under identical conditions. Our observations show that blocked arginine- and arginine-containing peptides bound more tightly to GAGs than the analogous lysine species and suggest that the difference was due to the intrinsic properties of the arginine and lysine side chains. The greater affinity of the guanidino cation for sulfate in GAGs is probably due to stronger hydrogen bonding and a more exothermic electrostatic interaction. This can be rationalized by soft acid, soft base concepts. PMID- 7487091 TI - Mechanism-based inactivation of cytochrome P450 2B1 by 9-ethynylphenanthrene. AB - The 7-ethoxycoumarin O-deethylase activity of rat cytochrome P450 (P450) 2B1 was inactivated by 9-ethynylphenanthrene (9EPh) in a time- and NADPH-dependent manner, and the loss of activity followed pseudo-first-order kinetics. At 20 degrees C, the extrapolated maximal rate constant of inactivation (kinactivation) was 0.45 min-1 and the inactivator concentration required for half-maximal inactivation (KI) was 138 nM. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) and HPLC analysis demonstrated that [2'-3H]-9EPh was irreversibly bound to the protein moiety of P450 2B1 and the stoichiometry of binding was determined to be 0.82 mol of inactivator bound per mole of P450 2B1. A radiolabeled peptide of approximately 3.0 kDa was identified by autoradiography after Tricine SDS-PAGE analysis of the peptides generated from a cyanogen bromide cleavage of [2'-3H]9EPh-inactivated P450 2B1. After HPLC separation of these peptides, the fraction containing the most radioactivity was analyzed by matrix assisted laser desorption ionization-mass spectrometry (MALDI-MS) and peaks at m/z 2720.9 and 2939.9 were detected. The lower mass peak represents the molecular ion (MH+) for the peptide Ile290 to Met314 (theoretical 2722.2), while the higher mass peak corresponds to the MH+ of the modified peptide (theoretical 2940.5). The difference in mass (approximately 219) would correspond to the addition of a phenanthrylacetyl group to the peptide. When the fraction containing the modified and unmodified peptides was further digested with pepsin and reanalyzed by MALDI MS, the site of attachment could be assigned to one of the amino acids contained in the peptide Phe297 to Leu307. PMID- 7487093 TI - Early increase in choline kinase activity upon induction of the H-ras oncogene in mouse fibroblast cell lines. AB - The effects of expression of the H-ras oncogene on phosphatidylcholine metabolism were examined in C3H10T1/2 and NIH3T3 cells expressing ras constitutively or under the control of inducible promoters. Cell lines expressing ras under the control of the mouse metallothionein promoter and the Escherichia coli lac operator/repressor system and an NIH3T3 cell line stably transfected with the ras oncogene were studied. Phosphocholine levels were elevated in the cells constitutively expressing ras and were increased 4-6 h upon induction in the inducible cell lines. Glycerophosphocholine, which is elevated five- to sixfold in constitutively transfected ras cells, did not increase at early times of induction, suggesting the absence of increased phosphatidylcholine degradation via a phospholipase A. Choline kinase activity increased within 4-6 h upon induction and correlated well with the increase in phosphocholine levels. This increase in phosphocholine levels could be prevented by the addition of hemicholinium-3, a competitive inhibitor of choline kinase. Expression of activated c-raf or v-raf also increased choline kinase activity, suggesting that the induction of choline kinase by ras is downstream of the ras/raf interaction. Long-term and short-term labeling experiments failed to detect evidence for increased phospholipase C activity. These results suggest that the increase in choline kinase activity observed in cells expressing ras is an early, integral part of ras transformation and is the main contributor to increased phosphocholine levels accompanying morphological changes. PMID- 7487092 TI - Mechanistic studies of 9-ethynylphenanthrene-inactivated cytochrome P450 2B1. AB - The mechanism of inactivation of the major phenobarbital-inducible cytochrome P450 of rat liver, P450 2B1, by 9-ethynylphenanthrene (9EPh) has been investigated. Matrix-assisted laser desorption ionization-mass spectrometry analysis of the cyanogen bromide-generated peptides from 9EPh-inactivated P450 2B1 confirmed the addition of a phenanthrylacetyl group to the peptide corresponding to residues 290 to 314. When this peptide was further digested with pepsin, the site of attachment could be assigned to one of the amino acids in the peptide Phe297 to Leu307 [Roberts, E. S., Hopkins, N. E., Zaluzec, E. J., Gage, D. A., Alworth, W. L., and Hollenberg, P. F. (1995) Arch. Biochem. Biophys. 323, 000-000]. The inactivation by 9EPh resulted in a 90-95% loss in the NADPH supported deethylation of 7-ethoxy-4-trifluoromethylcoumarin (EFC), but had no effect on the iodosobenzene- or cumene hydroperoxide-supported metabolism of EFC. The loss of NADPH-supported activity was not affected by the addition of cytochrome b5 or the presence of excess levels of reductase. The magnitude of the Type 1 spectral change upon the addition of benzphetamine was decreased with the 9EPh-modified protein. There was no decrease in the ability of modified 2B1 to form the steady-state level of the CO-reduced complex either enzymatically with NADPH and reductase or chemically with sodium dithionite, but the rate of reduction by reductase under anaerobic conditions was 57% that of native protein in the absence of substrate and 35% that of native protein in the presence of substrate. The 9EPh-modified 2B1 had an overall slower rate of NADPH oxidation, H2O2 formation, and formaldehyde formation during metabolism of benzphetamine compared to native 2B1. The ratio of H2O2 to HCHO was 1.0:1.0 for the native and 1.6:1.0 for the modified protein. The ability of the modified protein to form the steady-state level of the oxygen-iron complex in the presence of cyclohexane was decreased. These results are consistent with the idea that the covalent modification of one of the residues in the peptide Phe297 to Leu307 by the phenanthrylacetyl group impairs the reduction of P450 2B1 by reductase and also causes the uncoupling of NADPH utilization and oxygen consumption from product formation. PMID- 7487095 TI - Kinetic characterization of soybean seedling amine oxidase. AB - The kinetic characterization of soybean seedling amine oxidase (SSAO) and the effect of ionic strength and pH on the enzyme activity have been studied. A strong dependence of the activity on the carbon chain length of alpha-omega diamines, peaking at C8 and at C5, for Km and kc, respectively, was found. The analysis of ionic strength effects on activity showed a high sensitivity of Km and an insensitivity of kc to this parameter. This behavior and the different dependence of Km and kc on the carbon chain length suggest a two-stage equilibrium model for the oxidative deamination of polyamines by SSAO. The formation of the first intermediate (complex amine-enzyme) is controlled mainly by ionic interactions and by the structure of the polyamine, while the formation of the second intermediate depends only on the length of the carbon chain of the diamine. The molecular dissociation constants of the system soybean seedling amine oxidase-cadaverine were obtained from the dependence of kc, Km, and kc/Km on pH. A mechanism of the initial steps of reaction, involving a two-stage equilibrium model, is proposed. PMID- 7487094 TI - Self-diffusion monitors degraded cartilage. AB - This article demonstrates that both the bulk water self-diffusion coefficient (D) and the spatially resolved variation in D for lesion canine cartilage due to osteoarthritis is increased by about 25% over that of surrounding cartilage. This increase in D can be mimicked by enzymatic degradation of cartilage with trypsin, hyaluronidase, and collagenase, or by mechanical means. However, it is established here using excised disks of living cartilage whose proteoglycan and collagen contents were manipulated by biochemical intervention in tissue culture that the diffusion measurement is not sensitive to the proteoglycan content of cartilage. Instead, self-diffusion appears to monitor mesoscopic (nonspecific) tissue damage. These results show that D, measured in a spatially resolved manner by pulsed field gradient nuclear magnetic resonance imaging, can localize regions of cartilage degradation. PMID- 7487096 TI - Inhibition of neutrophil superoxide generation by hypericin, an antiretroviral agent. AB - We previously reported that phorbol 12-myristate 13-acetate (PMA)-induced superoxide (O2.-) generation of neutrophils was inhibited by hypericin, a photosensitizing pigment found in St. Johnswort (herb Hypericin triquetrifolium Turra), via a mechanism involving protein kinase C (PKC). To obtain further insights into the mechanism of inhibition, the effects of hypericin on stimulation-dependent O2.- generation and related enzymes of neutrophils were investigated. Hypericin inhibited O2.- generation of neutrophils induced by PKC dependent and -independent stimuli in a light- and concentration-dependent manner. Oxygen was required for the light-dependent inhibition by hypericin. NADPH oxidase activity in a cell-free system and TNF-alpha-induced tyrosyl phosphorylation of neutrophil proteins were also inhibited by hypericin in a concentration- and light-dependent manner. However, tyrosine kinase of p60src, an enzyme not bound to a membrane, was not inhibited either in the light or in the dark. Oxygen uptake of neutrophils by photosensitization with hypericin resulted in the formation of singlet oxygen (1O2), O2.-, and hydroxyl radical (.OH) and enhanced lipid peroxidation. The formation of 1O2 was inhibited by azide, a quencher of 1O2, but not by desferrioxamine (DSF), a ferric ion chelator. By contrast, both generation of .OH and lipid peroxidation were inhibited by DSF but not by azide. Furthermore, PMA-induced O2.- generation inhibited by hypericin partially recovered in the presence of azide but not DSF. These results suggested that the light-dependent inhibition of O2.- generation by hypericin might be due to inhibition of tyrosine kinase, PKC, and NADPH oxidase via an oxygen-dependent mechanism, possibly through both Type I and II photosensitization mechanisms. PMID- 7487097 TI - Ubiquinone-dependent recycling of vitamin E radicals by superoxide. AB - Superoxide-driven reduction of alpha-tocopheroxyl radical in the presence of ubiquinone-10 has been demonstrated in dimethylsulfoxide. Our HPLC measurements showed that ubiquinone-10 protected alpha-tocopherol against oxidation by KO2 in a concentration-dependent manner. alpha-Tocopherol was oxidized by KO2 to form ESR-detectable radicals of tocopherol ubisemiquinone. In the presence of ubiquinone-10, neither these radicals nor alpha-tocopheroxyl radicals (generated by uv light or PbO2) could be detected in ESR spectra. Instead, ESR signals of ubisemiquinone-10 radicals were observed. Formation of ubisemiquinone-10 radicals from ubiquinone-10 upon addition of KO2 was ascertained by their characteristic ESR and uv-vis spectra. alpha-Tocopherol caused a concentration-dependent decrease of the ubisemiquinone-10 radical steady-state concentration. We conclude that one-electron reduction of ubiquinone-10 by superoxide ion resulting in the formation of ubiquinone-10 radicals caused redox-cycling of alpha-tocopherol from its phenoxyl radical, thus preventing loss of alpha-tocopherol. This suggests that coenzyme Q may have another physiological function, i.e., protection of alpha-tocopherol against superoxide-driven oxidation. PMID- 7487098 TI - Cloning of a new pectate lyase gene pelC from Fusarium solani f. sp. pisi (Nectria haematococca, mating type VI) and characterization of the gene product expressed in Pichia pastoris. AB - Antibodies prepared against a pectate lyase (PLA) produced by a phytopathogenic fungus Fusarium solani f. sp. pisi (Nectria haematococca, mating type VI) were previously found to protect the host against infection. The cDNA and gene (pelA) for PLA were cloned and sequenced. A new pectate lyase gene, pelC, was isolated from a genomic library of F. solani pisi with pelA cDNA as a probe. A 1.3-kb DNA fragment containing the pelC gene and its flanking regions was identified and sequenced. The coding region of pelC was amplified by reverse transcription polymerase chain reaction using total RNA isolated from a pectin-induced F. solani pisi culture as template. The open reading frame of pelC was predicted to encode a 23.3-kDa protein of 219 amino acid residues, which shares 51% identity with PLA from F. solani pisi. No typical fungal leader peptide sequence could be identified at the N-terminus of the predicted protein sequence. The amplified pelC cDNA was expressed in Pichia pastoris yielding a pectate lyase C (PLC) with a molecular mass of 26.0 kDa and containing carbohydrates. PLC was purified to homogeneity using Mono Q anion-exchange chromatography. Purified PLC required Ca2+ for its activity and showed optimal lyase activity at pH 9.5 and 55 degrees C. Rapid drop in the viscosity of the substrate and Mono Q anion-exchange chromatography of the products generated by the lyase showed that PLC cleaved polygalacturonate chains in an endo fashion. Western blot using antibodies raised against PLA and PLC showed that PLC and PLA are immunologically related to each other. PMID- 7487099 TI - Analysis of the in vitro translation product of a novel-type Drosophila melanogaster aldolase mRNA in which two carboxyl-terminal exons remain unspliced. AB - Drosophila melanogaster generates three different types of aldolase mRNAs from a single gene by selective usage of the triplicate exons 4 (4 alpha, 4 beta, and 4 gamma), which encode three different isozymes having respective carboxyl termini. We have found the presence of a novel-type mRNA (named alpha beta) in which two final exons, 4 alpha and 4 beta, were retained unspliced. Herein, a cDNA clone containing the alpha beta sequence was inserted into pINIII and expressed in an Escherichia coli system. The product, which exhibited aldolase activity, was found to be isozyme alpha from the primary structure and the enzymological properties, with the 4 alpha sequence alone being present as the carboxyl terminus. In tissues of D. melanogaster, the production of mRNA encoding exon 4 alpha is known to be restrained to a low level. This may be understood by the fact that the aldolase gene of this species does not have a typical poly(A) signal at the 3' end in exon 4 alpha. Instead, the transcript-encoding exons, 4 alpha and 4 beta, might be produced when AATATA, which resides downstream of the coding frame in exon 4 beta, is recognized as a poly(A) signal during RNA processing. PMID- 7487100 TI - Human lactase-phlorizin hydrolase: evidence of dimerization in the endoplasmic reticulum. AB - Human lactase-phlorizin hydrolase [EC 3.2.1.23-3.2.1.62] is a disaccharidase located in the microvillus membrane of small intestinal epithelial cells. The enzyme is synthesized as a precursor protein in the endoplasmic reticulum and in addition to being glycosylated is subsequently proteolytically processed to the mature microvillus membrane-bound form after passing the trans-Golgi compartment. We studied the oligomerization of human lactase-phlorizin hydrolase in transfected polarized Madin Darby canine kidney cells using metabolic labeling and sucrose-density centrifugation analysis. We detected high mannose dimers of the lactase-phlorizin hydrolase precursor molecule after metabolic labeling with [35S]methionine at 37 and 15 degrees C. In addition, both complex-glycosylated lactase-phlorizin hydrolase precursor molecule and the mature microvillus membrane-bound enzyme showed this oligomeric structure. Chemical crosslinking resulted in the detection of covalently crosslinked lactase-phlorizin hydrolase dimers after sodium dodecyl sulfate polyacrylamide gel electrophoresis. These results provide evidence that oligomerization of lactase-phlorizin hydrolase is an early event and begins in the endoplasmic reticulum. PMID- 7487101 TI - Two related phenolic antioxidants with opposite effects on vitamin E content in low density lipoproteins oxidized by ferrylmyoglobin: consumption vs regeneration. AB - Endogenous alpha-tocopherol of low density lipoprotein (LDL) particles exposed to ferrylmyoglobin (iron in the form of FeIV = O) vanishes as a function of myoglobin concentration. After alpha-tocopherol depletion, subsequent heavy lipid peroxidation is prevented by caffeic and p-coumaric acids, i.e., phenolic acids present in foods and beverages, by a mechanism involving the one-electron transfer reaction between the phenols and the ferrylmyoglobin, with formation of metmyoglobin and the corresponding phenoxyl radicals from caffeic and p-coumaric acids, as previously discussed. Caffeic acid delays alpha-tocopherol consumption when present before oxidation challenging and restores alpha-tocopherol when added halfway during the reaction. Conversely, p-coumaric acid accelerates the rate of alpha-tocopherol consumption when added either before or during the oxidation reaction. In LDL enriched with alpha-tocopherol, caffeic acid induces an inhibition period of oxidation longer than that expected from the sum of discrete periods characteristic of the phenolic acid and alpha-tocopherol. Surprisingly, p-coumaric acid decreases the peroxidation chain rate. Similar effects of these phenolic acids on alpha-tocopherol consumption were observed in a Triton X-100 micellar system, i.e., in the absence of a peroxidation chain reaction. Results suggest that caffeic acid acts synergistically with alpha tocopherol, extending the antioxidant capacity of LDL by recycling alpha tocopherol from the alpha-tocopherol radical (i.e., alpha-tocopheroxyl radical). By contrast, the phenoxyl radical from p-coumaric acid (produced by electron transfer reaction between phenolic acid and ferrylmyoglobin) oxidizes alpha tocopherol. However, in spite of alpha-tocopherol consumption, the exchange reaction recycling p-coumaric acid can still afford an antioxidant protection to LDL on basis of the chain-breaking activity of p-coumaric acid. These results emphasize the biological relevance of small structural modifications of phenols on the interaction with alpha-tocopherol in LDL. The significance of these results in the context of atherosclerosis is discussed. PMID- 7487102 TI - Respiratory burst and tyrosine phosphorylation by vanadate. AB - We studied involvement of tyrosine-phosphorylated proteins in activation of NADPH oxidase in guinea pig neutrophils. Pervanadate, which is the oxidized form of orthovanadate, induced O2- production and protein tyrosine phosphorylation in neutrophils. O2- production induced by pervanadate was more sensitive to the tyrosine kinase-specific inhibitor, ST-638, as compared with the production induced by PMA. On the other hand, staurosporine more selectively inhibited PMA induced O2- production than pervanadate-induced production. These results indicate that tyrosine kinase, not protein kinase C, is involved in pervanadate induced O2- production. The tyrosine-phosphorylated proteins were detected in both the cytosol and membrane fractions prepared from pervanadate-induced neutrophils. In order to examine if tyrosine residues of some components of NADPH oxidase were directly phosphorylated, tyrosine-phosphorylated proteins were removed from solubilized membranes prepared from the pervanadate-stimulated neutrophils by immunoprecipitation with an anti-phosphotyrosine anti-body. NADPH oxidase activity in the solubilized membranes was not decreased by the treatment. These findings suggest that the components of NADPH oxidase are not tyrosine phosphorylated by pervanadate treatment, that tyrosine phosphorylation may be involved in the signal transduction pathway of NADPH oxidase activation by pervanadate, and that this pathway is independent of the activation by protein kinase C. PMID- 7487103 TI - Cloning and characterization of rat pancreatic beta-cell/liver type glucose transporter gene: a unique exon/intron organization. AB - Four overlapping lambda genomic clones encoding rat pancreatic beta-cell/liver type glucose transporter (GLUT2) have been isolated and characterized. The gene is about 35 kb long and contains 14 exons and 13 introns. Contrary to the exon 1 of the human or mouse counterpart, the rat GLUT2 gene has three additional noncoding exons which were identified by 5'-RACE and all four were designated exon 1a, 1b, 1c, and 1d. The intron sequences bordering the splice site junctions generally follow the GT/AG rule except for one intron which begins with GC. The exon sequences determined from genomic DNA sequencing showed some differences when compared to the published rat GLUT2 cDNA. Transcription initiation site was determined by primer extension and located 661 bp upstream of the ATG translation initiation codon. Several potential binding sites for transcription factors such as C/EBP, Sp1, AP1, HNF-5, and UPE were observed and they may be responsible for the regulation of GLUT2 gene expression. The promoter region of rat GLUT2 showed little homology when compared with those of human or mouse. However, striking sequence identity (84%) was found when the adjacent intron regions flanking exon 1c were compared with the -970/-721 region of the mouse GLUT2 promoter. A series of deleted mutant constructs of the putative promoter region linked to the CAT reporter gene showed promoter activity in the primary hepatocyte culture. The region containing -4542/+240 showed the highest CAT activity and further deletion of the region showed gradual decrease in CAT activity. PMID- 7487105 TI - Transcriptional regulation of rat hepatic low-density lipoprotein receptor and cholesterol 7 alpha hydroxylase by thyroid hormone. AB - Utilizing hypophysectomized and thyroidectomized rats, it is demonstrated that the rapid increase in expression of hepatic low-density lipoprotein (LDL) receptor and cholesterol 7 alpha-hydroxylase in response to thyroid hormone is due to increases in the rates of transcription of these genes. An increase in transcription of the hepatic LDL receptor gene was seen within 30 min after administration of triiodothyronine. The increase in transcription fully accounted for the increase in hepatic LDL receptor protein. Stimulation of transcription of these two genes provides for enhanced removal of LDL from serum and elimination from the body. PMID- 7487104 TI - cDNA cloning and baculovirus expression of the human liver endoplasmic reticulum P58: characterization as a protein disulfide isomerase isoform, but not as a protease or a carnitine acyltransferase. AB - The function of a 58-kDa liver microsomal protein (P58) is controversial. To help clarify the physiological function of this protein, particularly in humans, a full-length human liver cDNA clone was isolated, sequenced, and expressed in milligram quantities with the use of a baculovirus expression system. The deduced amino acid sequence of the mature protein contained two thioredoxin-like active site motifs (CGHC) and in its C-terminus a nuclear localization motif (KPKKKKK), and an ER-retention/retrieval motif (QEDL). The mature form of human P58 shared 95% amino acid sequence identity with the deduced amino acid sequences of a bovine liver cDNA, 93% with a murine B lymphocyte cDNA, and 91% with a rat basophilic leukemia cell cDNA. In contrast to reports on the activities of nonhuman forms of P58, the purified expressed human P58 showed no carnitine acyltransferase or protease activities. However, it did have protein disulfide isomerase activity, indicating that the physiological activity of human liver P58 may be attributed, at least in part, to this activity. PMID- 7487106 TI - Purification and characterization of procathepsin L, a self-processing zymogen of guinea pig spermatozoa that acts on a cathepsin D assay substrate. AB - Procathepsin L, the precursor to a powerful lysosomal cysteine proteinase, has been purified to apparent homogeneity from guinea pig spermatozoa, a novel and previously unrecognized source of this catalytically active zymogen. In the range of pH 5.0, procathepsin L (39,000 M(r)) readily self-processed yielding a mature, single-chain proteinase (29,000 M(r)) and an intact propeptide (10,000 M(r)) by what appeared kinetically to be an intramolecular reaction mechanism. These characteristics resembled those reported for the "major excreted protein" (MEP) of malignantly transformed mouse fibroblasts-a protein that has been characterized as the precursor to the mouse analog of human cathepsin L (B. R. Troen, S. Gal, and M. M. Gottesman (1987) Biochem. J. 246, 731-735). Other characteristics shared by the guinea pig and mouse zymogens included proteolytic activity at pH 5.0, homologous N-terminal amino acid sequences, and immunological relatedness. It was thus concluded that acrosomal procathepsin L is the guinea pig analog of MEP. Acrosomal procathepsin L had a specific activity on benzyloxy carbonyl-Phe-Arg-7-(4-methyl)coumarylamide (Z-Phe-Arg-NMec) of 30 mumol min-1 mg 1 enzyme at pH 3.2 and 37 degrees C. Relative to the assay substrate, rates on other fluorogenic substrates were 90% for Z-Phe-Cit-NMec, 63% for Z-Leu-Leu-Arg NMec, 43% for D-Phe-Ser(Bzl)-Phe-Phe-Ala-Ala-p-aminobenzoate (a "specific" cathepsin D assay substrate), and 32% for Z-Val-Val-Arg-NMec. No action was detected on Z-Arg-Arg-NMec or Arg-NMec. Mature cathepsin L showed the same relative order of substrate specificity as its proenzyme form, but the absolute rates were about 5-fold greater. Additionally, the mature (single-chain) form of cathepsin L displayed Km and kcat values on Z-Phe-Arg-NMec that yielded an exceptionally high catalytic coefficient (11,600 s-1 mM-1) compared to values reported for two-chain forms of cathepsin L. Self-processing by acrosomal procathepsin L at pH 5.5 was totally inhibited by leupeptin, cystatin C, Ep-475, and Z-Phe-Phe-CHN2 at 1 microM levels. Gossypol (0.1 mM) gave 94% inhibition. Interestingly, dextran sulfate (100 micrograms ml-1) gave a 3.6-fold increase in the rate of self-processing seen at pH 5.5--a phenomenon of potential physiological relevance in view of the high-negative-charge density present within the hyaluronic acid-rich outer layer (cumulus oophorus) of the ovum. PMID- 7487107 TI - The effects of an atpE ribosome-binding site mutation on the stoichiometry of the c subunit in the F1F0 ATPase of Escherichia coli. AB - We tested the hypothesis that the stoichiometry of the c subunit in the F0 sector of the Escherichia coli F1F0 ATPase is dependent upon the level of atpE gene expression. F0 was purified from cells carrying plasmids encoding the F0 subunits with and without a ribosome-binding site mutation preceding atpE, the gene which codes for the c subunit. Subunit-specific antibodies were used to quantitate the relative amounts of the b and c subunits. The decreased expression of atpE resulted in a significantly decreased amount of the c subunit in the purified F0. Immunoblot quantitation of the amounts of b and c subunits in F1F0 precipitated by anti-F1 antiserum also showed that the mutation produced significant differences in the stoichiometry of subunit c. The amount of c subunit assembled into the F1F0 synthesized from a plasmid carrying the atpE ribosome binding site mutation was 2-5 times less than the amount found in the F1F0 synthesized from a wild-type plasmid. Therefore, the stoichiometry of the c subunit assembled into the F1F0 complex appears to be variable, depending on the expression of atpE. PMID- 7487108 TI - Generation of superoxide and tyrosine peroxide as a result of tyrosyl radical scavenging by glutathione. AB - Enzymatically generated tyrosyl radicals are effectively scavenged by reduced glutathione (GSH), thereby generating glutathione thiyl radicals and superoxide radicals, subsequently. Here, we have used horseradish peroxidase to generate tyrosyl radicals and investigated the fate of the superoxide radicals. At low GSH concentrations (with a maximum effect at 250 microM) a major reaction was between superoxide and the phenoxyl radical leading to a tyrosine peroxide. Formation of the peroxide was confirmed using a peroxide-specific colorimetric assay and detection of a new HPLC-peak. Its formation was inhibited by superoxide dismutase (SOD). The peroxide decomposed slowly in a reaction that was accelerated by GSH to give a new chromatographic peak. Increasing the GSH concentration decreased the amount of tyrosine peroxide formed and caused increases in rates of oxygen uptake and GSH oxidation. These increases were not seen in the presence of SOD and are consistent with GSH scavenging superoxide and leading to oxygen-dependent chain oxidation of GSH. Both pathways are undesirable for the cell and are effectively suppressed only if GSH as a radical scavenger acts in concert with SOD. PMID- 7487109 TI - Comparison of isoniazid oxidation catalyzed by bacterial catalase-peroxidases and horseradish peroxidase. AB - The physical properties and activities of the purified catalase-peroxidase hydroperoxidase I (HPI) of Escherichia coli (EcHPI) and HPI with a carboxyl terminal extension of Mycobacterium tuberculosis (MtHPI-e) are compared to those of commercial preparations of horseradish peroxidase (HRP). The catalase peroxidase proteins had similar absorption spectra and differed primarily in that MtHPI-e has a higher peroxidatic to catalatic activity ratio than EcHPI. Trypsin cleavage of MtHPI-e resulted in the formation of an active catalase-peroxidase lacking the carboxyl-terminal extension. The three enzymes, HRP, MtHPI-e, and EcHPI, mediated the isoniazid- and H2O2-dependent production of radical species, as detected by nitroblue tetrazolium reduction. A constant flux of H2O2, generated in situ from glucose oxidase and glucose was used. MtHPI-e was more effective at isoniazid-dependent radical production than EcHPI and HRP. Similar qualitative results were obtained by staining nondenaturing polyacrylamide gels for activity with nitroblue tetrazolium in the presence of isoniazid and H2O2. The absorbance spectrum of HRP exhibited changes during incubation with isoniazid and H2O2 consistent with the formation of several typical reaction intermediates, whereas the catalase-peroxidases exhibited no distinct spectral changes. The results suggest that the sensitivity of M. tuberculosis to isoniazid may be the result of isoniazid-dependent radical formation by the catalase-peroxidase in the absence of other catalase activities to remove substrate H2O2. PMID- 7487110 TI - Bicyclomycin and dihydrobicyclomycin inhibition kinetics of Escherichia coli rho dependent transcription termination factor ATPase activity. AB - The primary site of action for the novel antibiotic, bicyclomycin, in Escherichia coli has been identified to be the rho transcription termination factor. The inhibition of rho poly(C)-stimulated hydrolysis of ATP by bicyclomycin has been found to proceed by a non-competitive, reversible pathway with respect to ATP (Ki = 20 microM). Inhibition by dihydrobicyclomycin was similar (Ki = 75 microM). No change in the inhibitory properties of the antibiotic was observed under the assay conditions with the two rho mutants, Cys202Gly and Cys202Ser, indicating that Cys-202 does not affect drug binding to rho. Prolonged incubation (32 degrees C, 12 h) of wild-type rho with bicyclomycin (20 mM) led to protein degradation and a slow, permanent loss of rho ATPase activity after dialysis. Evidence was obtained that trace amounts of proteases present with bicyclomycin were responsible for the observed protein degradation. Treatment of wild-type and mutant rho proteins with purified bicyclomycin (25 mM) led to approximately 80% loss of ATPase activity after dialysis with no apparent loss of protein. However, a reduction of the electrophoretic mobility of the bicyclomycin-treated rho versus wild-type rho was seen. Addition of either ATP or poly(C) to wild-type rho led to partial protection against bicyclomycin inactivation, while inclusion of both ligands provided near complete protection against inactivation. The observed loss of ATPase activity upon prolonged incubation of rho with excess purified bicyclomycin is attributed to the covalent modification of the protein by the antibiotic at multiple sites. PMID- 7487111 TI - Age-dependent association of isolated bovine lens multicatalytic proteinase complex (proteasome) with heat-shock protein 90, an endogenous inhibitor. AB - The multicatalytic proteinase complex (MPC) (proteasome) is a high-molecular weight proteolytic enzyme found in eukaryotic cells and archaebacteria. Regulatory proteins that inhibit or activate the MPC have been described. Association with an ATPase complex alters the specificity of the multicatalytic proteinase complex to permit cleavage of ubiquitinylated proteins. Unidentified proteins have been observed in highly purified preparations of the multicatalytic proteinase complex. Based on immunoreactivity and N-terminal sequencing, we have identified heat-shock protein 90 as a major component of the multicatalytic proteinase complex prepared from 1-month, but not 2-year bovine lenses. alpha Crystallin, a lens structural protein with chaperone activity, is also found in multicatalytic proteinase complex preparations. Both heat-shock protein 90 and alpha-crystallin inhibit hydrolysis of Cbz-Leu-Leu-Leu-MCA by the multicatalytic proteinase complex at a stoichiometry of 1 mol heat-shock protein per mole of MPC. Heat-shock proteins may interact with denatured proteins and facilitate their degradation. These studies give evidence for the involvement of heat-shock proteins in proteolysis by direct interaction with the multicatalytic proteinase complex. PMID- 7487112 TI - Role of cytochrome P450 in the metabolism and toxicity of hydroperoxides in isolated rat hepatocytes. AB - The contributions of cytochromes P450 (P450) to the metabolism and toxicity of hydroperoxides in freshly isolated rat hepatocytes were investigated utilizing 2,6-di-tert-butyl-4-hydroperoxy-4-methyl-2,5-cyclohexadienone (BHTOOH). This hydroperoxide was rapidly degraded in cell suspensions, and the products were identical to those determined previously with subcellular preparations of ferric P450. With 250 microM BHTOOH, the ratio of glutathione peroxidase-mediated:P450 mediated metabolism was estimated to be about 3:1. Surprisingly, BHTOOH was found to be a more potent cytotoxin than cumyl hydroperoxide (CuOOH), despite the fact that it caused substantially less lipid peroxidation than the latter. P450 inhibition enhanced the toxicity of BHTOOH, but lowered the toxicity of CuOOH. These data demonstrate that intracellular ferric P450 can compete with glutathione peroxidase to reduce hydroperoxides by 1- and 2-electron processes. If the alkoxy radical from homolytic cleavage of the O-O bond can undergo facile intramolecular reactions to nontoxic products, as with BHTOOH, the role of P450 is detoxification. On the other hand, if the alkoxy radical preferentially attacks membrane lipids, as with CuOOH, P450 contributes to lipid peroxidation and toxicity. It was determined that the levels of glutathione, protein thiols, and ATP decreased in parallel with BHTOOH-induced cell death, but no conclusions are possible concerning mechanisms underlying the relatively potent toxicity of BHTOOH. Toxicity may be related to the high lipophilicity of this hydroperoxide which, presumably, facilitates its passage into cells and distribution to various intracellular sites. BHTOOH appears to be an excellent model compound for investigating mechanisms of hydroperoxide-mediated cytotoxicity which do not involve lipid peroxidation. PMID- 7487113 TI - Differential regulation of manganese superoxide dismutase activity by alcohol and TNF in human hepatoma cells. AB - Increasing evidence implicates free radical processes in the pathogenesis of ethanol-induced liver injury. One of the antioxidant defense systems in mammalian cells is the mitochondrial enzyme manganese superoxide dismutase (MnSOD). MnSOD activity is increased by agents that cause oxidative stress. One such agent is the cytokine tumor necrosis factor-alpha (TNF). Increased serum/tissue TNF levels have been observed in alcoholic liver disease, and TNF has been postulated to play a role in ethanol-induced liver injury. Substantial evidence suggests that ethanol itself can cause oxidative stress. In order to investigate the mechanism of the cellular adaptive response to ethanol-induced oxidative stress, the effects of short-term ethanol exposure on MnSOD RNA, protein, and activity were determined in a human hepatoma cell line (HepG2). We found that exposure to ethanol (25 mM concentration) for 72 h increased the protein level and enzyme activity of MnSOD. However, examination of the mRNA levels of the enzyme showed no corresponding increase. Long-term administration of ethanol (10 weeks) did not significantly increase MnSOD protein and MnSOD activity. MnSOD activity was significantly increased by TNF. Thus it appears that both TNF and ethanol are capable of increasing MnSOD activity presumably via enhanced oxidative stress. However, unlike TNF, acute ethanol administration increases the activity of MnSOD without increasing MnSOD mRNA. The increase in MnSOD after a short-term dose of ethanol is diminished with repeated ethanol administrations. These findings are compatible with the view that chronic exposure to ethanol suppresses the cellular adaptive response to oxidative stress. If this adaptive response of MnSOD is lessened, it may have implications in the increased toxicity due to prolonged ethanol exposure. PMID- 7487115 TI - [Tumor rejection antigen and cancer immunotherapy]. AB - Tumor rejection antigen (TRA) recognized by cytotoxic T cells (CTL) have been identified on several murine tumors and human malignant melanomas. By utilizing those peptides as tumor vaccine, a new immunotherapy will be anticipated. Cancer cell vaccine and adoptive transfer of CTL/TIL have been undertaken successfully. Moreover, recombinant protein of tumor rejection antigen and heat shock protein emerge as promissing molecules as tumor antigen. To augument rather weak immune response, cytokine and gene therapies have now begun to be done. Cytokines can be used by themselves and also genes of cytokines can be introduced into either tumor cells or CTL/TIL. PMID- 7487116 TI - [Neoadjuvant chemotherapy and chemoradiotherapy in the treatment of esophageal cancer]. AB - Adjuvant therapy following surgery has been mainstream in surgical adjuvant therapy for the patients with esophageal cancer in Japan. In western countries, neoadjuvant therapy has become popular in which surgery is performed depending on the effects of preoperative treatment. Neoadjuvant chemotherapy offers the advantage of downstaging the primary tumor and enhancing resectability and the potential advantage of assessing the response to preoperative chemotherapy directly in the primary tumor. Disadvantages include possible emergence of chemotherapy-resistant tumor cells, as well as the delay in achieving effective local tumor control and postoperative morbidity. In the phase II studies, most regimens have included CDDP/5-FU. Pathological CR rates have been less than 10% and median survival terms from eight to 28 months. The rationale for the concurrent use of chemotherapy and radiotherapy is to combine an agent that has an effect upon systemic micrometastases with a modality that enhances local tumor control. In addition, a number of chemotherapeutic agents have radiosensitizing effects. The majority of trials have employed CDDP/5-FU combined with RT for a total dose of 30 Gy. Pathological CR rates were from 20 to 40% and median survival terms from 12 to 29 months. Neither neoadjuvant chemotherapy nor chemoradiotherapy increased operative morbidity mortality, and there was a statistically significant increase in survival in complete responders. However, though the early and median survival was improved, the cure rate was not. Both therapies remain investigational. PMID- 7487114 TI - The phosphorylation state of the cAMP response element binding protein is decreased in diabetic rat liver. AB - Phosphoenolpyruvate carboxykinase (PEPCK) is the rate-limiting enzyme of gluconeogenesis. This metabolically important enzyme is unique in that it has no known allosteric modifiers, and all of the regulation of its activity is exerted at the level of gene expression. The expression of the PEPCK gene in liver is elevated in most forms of diabetes, and plays a major contributory role in the hyperglycemia characteristic of this disease. In this study, we initiated studies to determine the molecular basis for the increased PEPCK gene expression in diabetes. RNase protection assays of RNA isolated from control, streptozotocin induced diabetic, and insulin-treated diabetic rat liver indicated that PEPCK mRNA levels are elevated two- to threefold in diabetic rat liver compared to controls. Nuclear run-on assays indicated that the increased PEPCK mRNA levels can be fully accounted for by changes in the transcription rate of the gene. We next initiated characterization of the cAMP response element binding protein (CREB) in diabetic rat liver, since it is known to play a major role in mediating the it is known to play a major role in mediating the basal transcriptional activity of the PEPCK gene as well as the cAMP-dependent stimulation of PEPCK gene transcription, the latter through the phosphorylation of serine 133 of CREB. Western blot analysis of nuclear lysates prepared from rat livers indicated that CREB protein levels in diabetic rat liver nuclei were similar to those of controls. However, using an antibody which specifically recognizes the serine 133 phosphorylated form of CREB, we found that the levels of phospho-CREB were significantly decreased in diabetic rat liver, an effect which insulin treatment reversed. This observation suggests that overexpression of the PEPCK gene in diabetes is not linked to the cAMP signaling system in liver. PMID- 7487118 TI - [Presence and future in neoadjuvant chemotherapy against gastric cancer]. AB - The present state and a future of neoadjuvant chemotherapy (NAC) was reviewed. Gastric cancer is classified as a cancer which is resistant against anti-cancer drugs. Consequently, surgery is the only method that offers hope of cure. Candidates for NAC are patients with stage IV disease, especially with T4 disease having no distant metastases. NAC is carried out for an increase of the resectability rate and the judgement of the in vivo chemosensitivity. Regimens with high fractional kill should be used, and and the tumor burden must be periodically estimated by CT scan, endoscopy and the serum levels of tumor markers. If the used regimen showed no response, a new protocol should be tried. Since a histological CR is very rare, not only primary tumor but metastases should be resected as much as possible. A completeness of the cytoreduction is a major factor for a long term survival. Surgery is carried out for the elimination of the drug-resistant cancer cells. Furthermore, early postoperative chemotherapy is very important for the improvement of survival after NAC. PMID- 7487119 TI - [Neo-adjuvant chemotherapy in patients with head and neck cancer]. AB - Standard treatment for head and neck carcinomas consists of surgery, and/or radiotherapy. However, standard treatment alone is less successful for advanced head and neck carcinomas. With the development of cisplatin (CDDP), the efficacy of chemotherapy including CDDP has markedly increased in head and neck carcinomas. Accordingly, neo-adjuvant chemotherapy has been applied for advanced carcinomas. The purpose of neo-adjuvant chemotherapy for head and neck carcinomas is to improve the curability by means of this chemotherapy strategy followed by standard treatment, to eradicate micro-metastatic lesions, and, more recently, organ preservation without compromising overall survival. The previous studies have demonstrated that this chemotherapy is efficacious for patients with complete response (CR), especially pathological CR, after chemotherapy in terms of preserving primary organ and overall survival. Therefore, the development of intensive chemotherapy regimen showing a high CR rate is necessary. Furthermore, the control of the secondary malignancies frequently occurring in patients with head and neck cancer, and the management of complications caused by aging and/or heavy drinking, are also indispensable to improve survival in advanced head and neck cancer patients. PMID- 7487117 TI - [Neoadjuvant therapy for colorectal cancer]. AB - To assess the role of neoadjuvant therapy in the treatment of colorectal cancer, we reviewed the literature investigating the effects of chemotherapy and/or radiotherapy given prior to surgery. Rationales for enteral administration of fluoropyrimidines preoperatively, in terms of pharmacokinetic and antitumor effects, have been demonstrated in clinical studies. For instance, the concentration of 5-FU in cancer tissue was shown to be significantly higher than in normal tissue, and a dose-response relationship between the cumulative doses of the agent and the histological antitumor effect was observed. Among several comparative studies, there were reports suggesting the efficacy of preoperative chemotherapy. The treatment group had a slight, but insignificant, improvement in disease-free interval or long-term survival. On the other hand, some randomized trials for rectal cancer demonstrated that preoperative radiotherapy could decrease local failure, and that patients who underwent curative surgery appeared to have an improved 5-year survival compared to the controls, although overall survival was not improved. Recent reports on combined modality treatment with chemotherapy and radiotherapy clearly showed a downstaging effect on the primary tumor and the decreased prevalence of regional involved lymph node. Further developments through rationally designed study are necessary to ascertain the efficacy of neoadjuvant therapy for colorectal cancer. PMID- 7487120 TI - [Preoperative chemotherapy in genitourinary cancer]. AB - Preoperative chemotherapy is the standard therapy for metastatic testicular cancer, and is also a candidate for a new treatment modality in management of bladder cancer. Since the advent of cisplatin-based chemotherapy in the late 1970s, a majority of metastatic testicular cancer patients can be made disease free and cured with chemotherapy followed by surgery. The development of combination chemotherapy including cisplatin and methotrexate in the late 1980s also improved the tumor regression rate for bladder cancer. However, the chemotherapy for the metastatic bladder cancer can not control a relapse after initial response, nor improve the survival rate, significantly. While several pilot studies suggest that the neoadjuvant chemotherapy may be beneficial in non metastatic invasive bladder cancer, the results of randomized trials are presently not available to define the role of this new treatment modality in management of bladder cancer. Intra-arterial chemotherapy combined with radiotherapy and conservative surgery may be indicated for selected patients as bladder preservation therapy. PMID- 7487122 TI - [Augmentation of chemotherapeutic efficaciousness of UFT by oral l-leucovorin--l leucovorin factors enhancing cytotoxic activity of 5-fluorouracil]. AB - The relationship between augmentation of cytotoxic activity of 5-fluorouracil (FUra) by l-leucovorin (l-LV) and combination schedule was investigated using human colon adenocarcinoma KM 20 C cell line. The enhancement of cytotoxic activity of FUra by l-LV or 5-CH3FH4, a main metabolite after oral administration of l-LV, was observed after 96-hr exposure to either l-LV or 5-CH3FH4 at concentrations of greater 0.001 microM or 0.3 microM, respectively. We found that 1.5 microM FUra, which has no effect on cell-growth without l-LV, could suppress strongly the proliferation if cells were co-treated with l-LV for more than 8 hr. Therefore, enhancement of cytotoxic activity of FUra by l-LV depended on the time cell exposure to l-LV. Moreover, this effect was observed even at a concentration of 0.01 microM l-LV after 48-hr exposure. Likewise, it is known that the antitumor activity of FUra correlates with the time-schedule of treatment with FUra. These results suggest that in addition to FUra derivatives which continuously release FUra, it is beneficial for the combination therapy of FUra with l-LV to consider continuous administration of l-LV as well. PMID- 7487121 TI - [Augmentation of chemotherapeutic efficaciousness of UFT by oral l-leucovorin- growth-inhibitory activity of combination against human tumor xenograft]. AB - Combination chemotherapy with FUra and LV has been reported as a useful treatment for patients suffering from colon carcinoma. Usually, both FUra and LV are administered by intravenous infusion, but not orally. UFT, an anti-neoplastic agent consisting of FT and uracil, is widely used for oral administration in Japan. Using human tumor xenografts of 10 cell lines, we evaluated the efficacy of UFT combined with l-LV, which is the active form of LV, by oral administration. Combined treatment of UFT with l-LV was more effective than UFT alone on the growth suppression of colon carcinoma (KM 20 C, Col-1) and mammary carcinoma (H-31, MX-1). When 1.85 mg/kg (5.55 mg/m2) of LV was given to tumor bearing mice, the antitumor activity of UFT was augmented and at a dose of 5.56 mg/kg (16.7 mg/m2) of LV, it was significantly augmented. Among various 5-FU derivatives, such as UFT, 5'-DFUR or FUra, combined treatment using UFT with l-LV was the most effective by oral administration. l-LV did not improve the anti tumor efficacy or toxicity of 5'-DFUR. l-LV seemed to augment the anti-tumor activity of FUra, but not significantly. These results suggest that combination chemotherapy of UFT with LV is a promising approach for the clinical treatment of human colon cancer. PMID- 7487123 TI - [Development of new in vitro chemosensitivity test using collagen gel droplet embedded culture and its clinical usefulness]. AB - We developed a new in vitro assay for chemosensitivity test using collagen gel droplet embedded culture and image analysis. In this in vitro assay, we successfully minimized the cancer cell number required for culture to approximately 3-10 x 10(3) cells for each 30 microliters collagen gel droplet, obtained the sufficient growth of cancer cells using serum-free medium while suppressing the growth of fibroblastic cells, and measured the volume of cancer cells by eliminating the contaminating fibroblastic cells by an image processing technique. Anticancer effects of the in vitro assay showed a very good correlation with those of in vivo nude mouse assay using human cancer cell lines. The success rates of the in vitro assay for 141 surgical specimens of primary lung cancers and for 65 of primary breast cancers were 89 and 80%, respectively. The accumulated in vitro assay response rates of MMC, CDDP, VDS and VP-16 for primary lung cancers and of MMC, 5-FU and ADR for primary breast cancers were similar to the respective clinical response rates. These results suggest that this in vitro chemosensitivity test may be practically useful for clinical applications. PMID- 7487124 TI - [Usefulness of 123I-MIBG myocardial SPECT in patients with hematologic malignancies with chemotherapeutic agent-induced cardiomyopathy]. AB - In 59 patients with hematologic malignancy undergoing chemotherapy regimens including adriamycin (ADR), myocardial imaging was performed by SPECT with 123I metaiodobenzylguanidine (MIBG) to detect cardiac sympathetic nerve abnormalities. SPECT imaging was performed 20 min and 4 hr after intravenous injection of 123I MIBG, and the washout rate (WR) for the entire left ventricle was calculated. Then the relationship of the WR to the total dose of ADR, the left ventricular ejection fraction, and the frequency of arrhythmias was evaluated. It was found that the WR was related to the total dose of ADR, suggesting that it is a useful parameter for cardiac sympathetic nerve damage. As the WR increased, the frequency of ventricular arrhythmias also increased. The WR was usually normalized 3-6 months after the discontinuation of ADR. In patients with a WR > or = 50%, however, prolonged follow-up was required. In conclusion, the WR on 123I-MIBG myocardial SPECT scans may be useful as a parameter for predicting cardiac toxicity requiring the discontinuation of ADR or dose reduction. PMID- 7487126 TI - [Induction chemotherapy followed by adjuvant surgery (IC-AS) in patients with stage I-II small cell lung cancer (SCLC)]. AB - Ten patients with stage I-II SCLC received IC-AS between 1984 and 1993. As induction chemotherapy, COMP-VAN alternating chemotherapy and CAV-PVP hybrid chemotherapy were administered. The former consisted of a 4-drug combination of cyclophosphamide (CPA), vincristine (VCR), methotrexate (MTX) and procarbazine alternated with a 3-drug combination of etoposide (ETP), adriamycin (ADM) and nimustine every 4 weeks. In the latter, a 3-drug combination of CPA, ADM and VCR given on day 1, and a 2-drug combination of ETP and cisplatin on day 8, were repeated every 4 weeks. All the patients had an objective response, including one complete response by induction chemotherapy. Post-operative pathology revealed SCLC in 4 patients, adenocarcinoma in 2 and no tumor (pathological CR) in 4. Four patients relapsed, and a intrathoracic relapse was experienced in only 2 patients. Six patients have died: 3 from relapsing SCLC, 2 from stomach cancer, and 1 from squamous lung cancer, who was salvaged from relapsing SCLC. The median survival time was 27.5 months, and the 3-year survival rate 37.5%. These results indicate that IC-AS is highly effective for stage I-II SCLC and warrant additional studies comparing IC-AS with chemo-radiotherapy. PMID- 7487125 TI - [UFT/CDDP preoperative chemotherapy for progressive gastric cancer--histological antitumor effects and thymidylate synthase inhibition rate]. AB - Pre-operative chemotherapy with concomitant use of UFT and CDDP (UFT: oral administration of 400 mg/day for 2 weeks till one day before operation, CDDP, one intravenous drip of 40 mg/m2 one week before operation) was used for 24 untreated cases of advanced stomach cancer diagnosed as resectable pre-operatively, and the histological antitumor effect analyzed in dissected preparation and the thymidylate synthase inhibition rate (TSIR: %) in tumor tissue were examined. The average administration dose of CDDP was 61.1 mg/body, and the average total administration dose of UFT was 5.0 g/body. The histological antitumor effect was grade 0 in 8 cases (33.3%), grade 1a in 10 cases (41.7%), grade 1b in 5 cases (20.8%), and grade 2 in 1 case (4.2%). TSIR in tumor tissue was under 10% in 2 cases (9.1%); over 10% and under 20% in 4 cases (18.2 %); over 20% and under 30% in 6 cases (27.3%); over 30% and under 40% in 5 cases (22.7%); over 40% and under 50% in 3 cases (13.6%); over 50 % in 2 cases (9.1%), and not measurable in 2 cases, with the average of 29.0%. The correlation was observed between histological anti-tumor effect and TSIR in tumor tissue (p < 0.05). These results suggest the possibility that the anti-tumor effect can be estimated at the in vivo level from measurement of TSIR in tumor tissue. PMID- 7487129 TI - [A case of unresectable advanced gastric cancer responding remarkably to combined chemotherapy with 5-fluorouracil and low-dose cisplatin]. AB - A 62-year-old male patient complaining epigastralgia was diagnosed as having Borrmann type 2 gastric concer (por). The primary lesion was unresectable, so the patient was treated by combined chemotherapy with 5-fluorouracil and low-dose cisplatin for 4 weeks, which resulted in the disappearance of primary tumor and a remarkable reduction of metastatic lymph nodes. The patient has a good quality of life without any sign of recurrence now. PMID- 7487128 TI - [A case of gastric cancer responding well to MTX/5-FU (methotrexate/5 fluorouracil) and PMUE (CDDP, MMC, UFT, etoposide) therapies upon lymphnode recurrence]. AB - We encountered a case of non-curatively resected gastric cancer (p1, n4) who responded well to sequential MTX/5-FU therapy and PMUE therapy. A 63-year-old man was admitted to our hospital with complaints of nausea and vomiting. Upper gastrointestinal examination and CT scan revealed Borrmann type 3 gastric cancer with pyloric stenosis and multiple paraaortic lymphnodal metastasis. The patient underwent palliative gastrectomy for extensive gastric cancer (H0, P1, N4, T3, Stage IV b). Histological examination of the resected stomach revealed poorly differentiated adenocarcinoma with paraaortic lymphnodes metastasis (n4) and peritoneal dissemination (p1). Chemotherapy with sequential MTX/5-FU was given 13 times. Ten months after the operation, abdominal pain and back pain required analgesic treatment. Abdominal CT scan revealed increased size of paraaortic lymphnodes, suggesting recurrence. Sequential MTX/5-FU therapy was switched by PMUE therapy. Lymphnode size became smaller and habitual analgesics could be discontinued. Since then he was given MTX/5-FU and PMUE therapies alternately on an ambulant basis. The patient resumed his daily activities at 2 years and 8 months after the operation. PMID- 7487127 TI - [A randomized crossover comparison of azasetron and granisetron in the prophylaxis of emesis induced by chemotherapy including cisplatin]. AB - The clinical application of 5-HT3 receptor antagonists has enabled continuation of the course of chemotherapy including cisplatin, which induces strong nausea and vomiting, and to prevent the delay of curative treatment for cancer patients receiving neoadjuvant chemotherapy. However, with the development of basic research on the mechanisms of vomiting, each 5-HT3 receptor antagonist has appeared to have different pharmacological actions and, subsequently, the difference in the clinical efficacy of each drug has been reported in Europe and USA. In freshly advanced head and neck carcinoma cases, a randomised crossover study was performed to compare the efficacy and safety profile of a single intravenous dose for 7 days of azasetron (10 mg/day) or granisetron (3 mg/day) in the prophylaxis of nausea and vomiting induced by multi-drug chemotherapy including cisplatin (50 mg/m2 or 60 mg/m2). Anti-emetic effects were evaluated by the protective rates for nausea and vomiting for 7 days following the start of cisplatin administration. Both 5-HT3 receptor antagonists were highly effective in the prophylaxis of acute and delayed emesis induced by chemotherapy, whereas the efficacies of azasetron on day 3 and 4 were superior to those of granisetron. No adverse effect of either drug was observed in this study. PMID- 7487130 TI - [A case of advanced esophageal cancer made resectable by preoperative combination therapy with 5-FU and CDDP]. AB - The authors experienced a case of advanced esophageal cancer made resectable by combination therapy with 5-FU and CDDP as a neoadjuvant chemotherapy. The patient was a 69-year-old-man suffering from esophageal cancer of A3.N4 (+).Pl0.M0 at stage IV. At this case was diagnosed to be radically unresectable, this form of combination therapy was used. The patient showed PR after 2 courses and the operation could then be conducted. The intraoperative findings revealed fibrous fusion of tumor with the aorta, but no direct invasion. The metastatic lymph nodes were necrotized and reduced. It was evaluated as Grade 3 in accordance with the "Histologic criteria for the effects of anticancer chemotherapy." The postoperative course was favorable without recurrence. This therapy caused no adverse reactions and seems effective as a neoadjuvant chemotherapy for advanced esophageal cancer. PMID- 7487131 TI - [A case of pancreatic cancer surviving for 7 years 11 months following intra arterial and portal infusion chemotherapy]. AB - A case of unresectable pancreatic cancer in a 76-year-old female was reported, in which a tumor had liver involvement. She was treated with infusion chemotherapy of mitomycin C (MMC) from gastroduodenal artery and portal vein after palliative operation. A drug was administered via an extracorporeal infusion catheter. MMC was given totally 47 mg from artery and 27 mg from portal vein for 9 months. Then, the serum CA 19-9 level gradually decreased, the metastatic liver tumor disappeared and the pancreatic tumor became smaller. After that, the tumor did not grow. The patient survived for 7 years 11 months until she died from other causes. Thus, it seems that infusion chemotherapy might be tried in some cases of pancreatic cancer. PMID- 7487132 TI - [Five-day continuous intravenous infusion of cisplatin and etoposide for extensive stage small cell lung cancer]. PMID- 7487133 TI - [Introduction of a new aromatase inhibitor fadrozole hydrochloride hydsate]. AB - Fadrozole (CGS 16949 A, brand name: Afema) is an aromatase inhibitor developed firstly in Japan. This compound reduces estrogen levels in the body after administration, suppressing the growth of breast cancer. In animal experiments, this showed an inhibitory activity in vivo against estrogen-depended mammary tumor and the effect was potentiated by the combination of tamoxifen cytrate. In the domestic clinical trials against post-menopausal advanced-recurrent breast cancer, irrespective of the cases with ER positive or negative and even including pretreated cases, the compound showed response rate of 19.3%, the rate of long NC of 18.2%, and the total response rate of 37.5%. The prognosis of NC cases was similar to the effective (CR + PR) cases. The median survival time was 323.5 days which were better than the previous endocrine therapies. The side effects mainly consisted of nausea, vomiting, loss of appetite, abdominal pain, and fatigue, but these were all the level of grade 1. This compound-seemed to be a promising drug for the treatment of patients with post-menopausal breast cancer. PMID- 7487134 TI - [Prognostic factors of prostatic cancer]. AB - In the past ten years, the treatment of prostate cancer has undergone remarkable changes. Furthermore, systematic prostate biopsy under transrectal ultrasonography or the measurement of prostate specific antigen has provided significant improvement in the detection of prostate cancer. Considering the increasing detection of prostate cancer, it is very important to clarify prognostic parameters in order to select the optimal treatment of given patients with localized or advanced prostate cancer. Recent statistical analyses showed that tumor volume, lymph node involvement and Gleason score would be significant prognostic indicators in localized prostate cancer. In advanced cancer, the response to the initial endocrine therapy might be a good prognostic parameter in addition to the histopathological features of primary tumors. In order to determine the factors which predict the prognosis of prostate cancer more accurately, the molecular biological approach is now ongoing, and may provide novel parameters for the decision making in the treatment of prostate cancer. PMID- 7487135 TI - Proliferation and interferon-gamma receptor expression in psoriatic and healthy keratinocytes are influenced by interactions between keratinocytes and fibroblasts in a skin equivalent model. AB - Epidermal-dermal interactions were studied in a skin equivalent model. Six combinations of keratinocytes and fibroblasts from healthy and psoriatic skin were used. TPA (12-O-tetradecanoylphorbol-13-acetate) was used to determine whether the expression of the IFN-gamma receptors in keratinocytes was related to epidermal differentiation and proliferation. These phenomena were assessed by immunohistochemistry. In all epidermal outgrowths, the epidermal growth factor receptor was expressed throughout the epidermis, cytokeratin 16 suprabasally, and filaggrin and involucrin in its superficial part. The IFN-gamma receptor was expressed throughout the epidermis, but was unevenly distributed. The expression of the IFN-gamma receptor was quantified by confocal laser scanning microscopy both in the whole of epidermis and in areas with the strongest intensity. The total amount varied to a minor degree in the epidermal outgrowths of different origins and was unaffected by TPA. In high-intensity areas interactions between keratinocytes and fibroblasts did influence the amount of IFN-gamma receptor expression and TPA decreased the expression by 13%. There was no correlation between the proliferation rate and the expression of the IFN-gamma receptor. Psoriatic and healthy keratinocytes were equally well differentiated in the skin equivalents. The interferon-gamma receptor was similarly expressed under these conditions. The growth rate, assessed by Ki-67-positive nuclei in the basal layer, was highest in healthy keratinocytes. Keratinocytes from psoriatic lesions increased their growth rate when cocultured with psoriatic fibroblasts compared with normal ones, indicating that fibroblasts may be of importance for epidermal hyperproliferation in psoriatic lesions. PMID- 7487136 TI - Murine epidermal Langerhans cells express CD48, which is a counter-receptor for mouse CD2. AB - It has recently been demonstrated that CD48, which is expressed on T cells, B cells, thymocytes and splenocytes, is a ligand for mouse CD2 and that it can function as one of the costimulatory molecules in the activation of T cells. In this study, we examined the expression of CD48 on epidermal Langerhans cells (LC), which are potent antigen-presenting cells in the skin. Both freshly isolated and short-term-cultured LC were shown to express CD48 by flow cytometry. In contrast to most of the adhesion molecules expressed on LC, CD48 expression on short-term-cultured LC did not differ significantly from that on freshly isolated LC. We also examined the contribution of CD48 to antigen presentation by LC. We stimulated the myoglobin-specific T-cell clone, TK.G4, and allogeneic splenic T cells with freshly isolated LC and cultured LC, respectively, in the presence of various concentrations of anti-CD48 monoclonal antibody (mAb). Even at the concentration of 30 micrograms/ml, however, the anti-CD48 mAb did not show any inhibitory effects on either allogeneic or antigen-specific T-cell proliferation, whereas at a concentration 10 micrograms/ml, the anti-CD48 mAb significantly suppressed the proliferation of spleen cells stimulated with phytohaemagglutinin (PHA). These findings show that LC persistently express CD48, although its direct role in antigen presentation has not yet been clarified in vitro. PMID- 7487137 TI - Epidermal melanocytes in normal and tyrosinase-negative oculocutaneous albinism fetuses. AB - In tyrosinase-negative (type IA) oculocutaneous albinism (tyr(-) OCA) fetuses the development of melanocytes has not been fully elucidated. We analysed the distribution of melanocytes in skin samples from a fetus with tyr(-) OCA and from four normal fetuses. Skin samples obtained from 12 different body sites of each fetus were examined by transmission electron microscopy, an electron microscopic DOPA reaction test and immunohistochemistry. No S100 protein-positive cells were detected in any sample. There were fewer HMB-45-positive melanocytes in the skin of the tyr(-) OCA fetus than in the skin of normal fetuses from all body sites sampled. The greatest number of HMB-45-positive melanocytes was present in samples from the scalp of the normal fetuses and in those from the lower limbs of the tyr(-) OCA fetus. Very few melanocytes were detected immunohistochemically in samples from the soles and palms, though their presence was confirmed by transmission electron microscopy. The electron microscopic DOPA reaction test enhanced the melanization of melanocytes in samples from the normal fetuses but not in those from the tyr(-) OCA fetus. Postembedding immunogold electron microscopy using the HMB-45 antibody revealed that the HMB-45 antigen was localized mainly on stages I and II melanosomes. The presence of epidermal melanocytes in samples from all fetal body areas obtained at 17-21 weeks of gestation justifies the use of the electron microscopic DOPA reaction test in the prenatal diagnosis of tyr(-) OCA. PMID- 7487138 TI - A fibroblast cell line cultured from a hypertrophic scar displays selective downregulation of collagen gene expression: barely detectable messenger RNA levels of the pro alpha 1(III) chain of type III collagen. AB - The present study was designed to investigate the expression of type I, III and VI collagens by a fibroblast cell line initiated from a hypertrophic scar. The same tissue has previously been demonstrated to display markedly elevated expression of type I and III collagen mRNAs in vivo. Unexpectedly, slot-blot and Northern hybridizations revealed a barely detectable steady-state level of pro alpha 1(III) collagen chain mRNA in cultured hypertrophic scar fibroblasts. The levels of pro alpha 1(I) and alpha 2(VI) collagen chain mRNAs were essentially the same in fibroblasts cultured from hypertrophic scar and in fibroblasts cultured from normal skin. However, Northern blot analyses indicated that the ratio of 5.8 kb to 4.8 kb species of pro alpha 1(I) collagen mRNA was slightly reduced in fibroblasts originating from the hypertrophic scar compared to that in normal fibroblasts. When normal fibroblasts were incubated in conditioned medium from hypertrophic scar cultures, the expression of pro alpha 1(III) collagen chain mRNA decreased to a markedly lower level. Our studies suggest that collagen synthesis by fibroblasts in hypertrophic scars is stimulated by humoral factors which are active only in vivo. Furthermore, the results suggest that fibroblasts cultured from hypertrophic scar display a selective downregulation of different collagen genes and that this downregulation is exerted through an autocrine mechanism. PMID- 7487140 TI - Secretory component (polymeric immunoglobulin receptor) as an intrinsic inhibitor of biological functions of interferon gamma in keratinocytes. AB - Secretory component (SC) acts as a transmembrane polymeric immunoglobulin receptor of epithelial cells and is known to bind to polymeric IgA and to contribute to the secretion of secretory IgA (sIgA). We describe a new biological function for free SC (FSC) by which the expression of intercellular adhesion molecule-1 (ICAM-1) and HLA-DR induced by interferon gamma (IFN gamma) is inhibited in human keratinocytes. This activity coincided with suppression of adenosine cyclic 3,5-monophosphate (cyclic AMP) production in keratinocytes. Keratinocytes produced SC after stimulation with IFN gamma and this production was suppressed by the addition of H-7 or propranolol. The addition of propranolol resulted in prolongation of ICAM-1 expression on keratinocytes induced by IFN gamma. These results suggest that endogenously produced SC, as well as exogenously added FSC, acts as an inhibitor of IFN gamma. Therefore, our results suggest that SC plays an antiinflammatory role in the pathogenesis of inflammatory skin diseases via inhibition in keratinocytes of IFN gamma induced expression of ICAM-1 and HLA-DR. PMID- 7487139 TI - Prostaglandin I1 analogues, SM-10902 and SM-10906, affect human keratinocytes and fibroblasts in vitro in a manner similar to PGE1: therapeutic potential for wound healing. AB - The newly synthesized prostaglandin (PG) I1 analogues, SM-10902 and SM-10906, were compared with PGE1 in terms of their biological effects on cultured normal human keratinocytes (NHKs) and human dermal fibroblasts (HDFs) in order to evaluate their therapeutic potential for cutaneous wound healing. The PGI1 analogues had a direct effect on cell proliferation of HDFs as did PGE1, but inhibited cell growth of NHKs in contrast to the stimulatory effect observed with PGE1. In contrast to NHKs stimulated with PGI1 analogues, which exhibited low levels of adenosine 3,5-cyclic monophosphate (cAMP). HDFs stimulated with these analogues responded in a dose-dependent manner with extremely high levels of cAMP. Conditioned media (CM) derived from media in which HDFs had been incubated with both the PGI1 analogues promoted NHK proliferation. HDF production of interleukin (IL)-6 increased in response to the PGI1 analogues. Since IL-6 was shown to promote cell growth of NHKs, enhancement of NHK proliferation by CM was thought to be due to IL-6 derived from HDFs stimulated with the PGI1 analogues. PMID- 7487141 TI - Histopathological and immunohistochemical changes in psoriatic skin during peptide T treatment. AB - Ten patients with plaque-type psoriasis were treated with 2 mg peptide T i.v. for 28 days. Six patients responded with a substantial clinical improvement. Sequential biopsies from skin lesions were taken before, during and after treatment. The histological score (defining the activity of the psoriasis), the epidermal thickness and the number of infiltrating dermal lymphocytes were all reduced in the six patients who responded to the treatment. An increase in the number of CD1+ dendritic cells was detected immunohistochemically in the epidermis of the responders. The nonresponders did not display any pronounced changes. PMID- 7487142 TI - FK506: therapeutic effects on lupus dermatoses in autoimmune-prone MRL/Mp-lpr/lpr mice. AB - The effects of FK506, a new immunosuppressive agent, on the development of lupus dermatoses were investigated in the autoimmune-prone MRL/Mp-lpr/lpr (MRL/lpr) mouse, which is an animal model for the spontaneous development of skin lesions similar to those of human lupus erythematosus (LE). FK506 reduced the incidence of skin lesions, lupus nephritis, the titre of serum anti-double-stranded DNA antibodies and the massive T cell proliferation. The incidence and magnitude of IgG deposition at the dermoepidermal junction were not changed. These results suggest that FK506 is a promising immunosuppressive agent for the control of autoimmune skin diseases. PMID- 7487144 TI - A local lymph-node assay validation study of a structure-activity relationship model for contact allergens. AB - A structure-activity relationship model for prediction of contact allergenic potential of chemicals had previously been developed. The model had been shown to be able to classify known allergens and nonallergens using data on physicochemical and reactivity parameters of functional groups by discriminant two-value multiple regression analysis. To investigate the model, six selected chemicals which had not been previously investigated for allergenicity were studied with both the model and a murine local lymph-node assay. The same compounds were predicted to be allergens (3-bromo-2-coumaranone, 1 nitrocyclohexene and alpha-acryloyloxy-beta, beta-dimethyl-beta-butyrolactone) and nonallergens (1-carbethoxy-4-piperidone, 6,7-dimethoxy-2-tetralone and 9 acetylanthracene) by both the model and the local lymph-node assay. PMID- 7487143 TI - Treatment of contact hypersensitivity with urocanic acid. AB - In order to investigate the effect of cis-urocanic acid (UCA) on a delayed-type hypersensitivity response in humans, a contact hypersensitivity reaction was induced on four test sites on the back of 33 volunteer subjects. The first test site was pretreated with cis-UCA immediately before application of the allergen. The second and third test sites were posttreated on the second and third days of the hypersensitivity response with cis-UCA and a class III corticosteroid, respectively. The fourth test site was used as a positive control. The cutaneous blood flow of the test sites was measured using laser Doppler flowmetry. Pretreatment with cis-UCA reduced the hypersensitivity response significantly. It is possible that cis-UCA could be used in the preventive treatment of contact hypersensitivity responses. PMID- 7487145 TI - Itch and inflammation induced by intradermally injected interleukin-2 in atopic dermatitis patients and healthy subjects. AB - To explore the pruritogenic and inflammatory effects of cytokines, a single dose of 20 micrograms recombinant human interleukin-2 was injected intradermally into eight patients with atopic dermatitis and eight healthy controls. The study was double-blind and randomized with glucose as a negative control. The effects were evaluated by recording local itch and erythema over 72 h and by examining skin biopsies taken at 24 h and 72 h. In patients and controls, interleukin-2 provoked a low-intensity local itch with maximal intensity between 6 h and 48 h and erythema with maximal extension between 12 h and 72 h. In the atopic dermatitis patients, these reactions tended to appear earlier and were less pronounced than in the healthy controls. Interleukin-2 induced dermal mononuclear cell infiltrates consisting mainly of CD3+ cells. A majority of the T cells were CD4+. The number of dermal CD25+, HLA-DR+ and ICAM-1+ cells was also increased at the interleukin-2 induced spongiosis and exocytosis as well as HLA-DR+ and ICAM-1+ keratinocytes. The microscopic findings tended to be more prominent at 72 h than at 24 h in both groups, but with a somewhat slower onset in the atopic dermatitis patients. In conclusion, a single intradermal injection of interleukin-2 induced local itch, erythema, dermal T-cell infiltrates, spongiosis, exocytosis and activation of keratinocytes both in atopic dermatitis patients and in healthy controls. PMID- 7487146 TI - Autoantibodies to the heat-shock protein hsp73 in localized scleroderma. AB - We determined the presence of antibodies to the heat-shock protein hsp73 (anti hsp73) in 57 serum samples from patients with localized scleroderma using an enzyme-linked immunosorbent assay (ELISA). In addition, 30 samples from healthy individuals, 30 from patients with systemic lupus erythematosus (SLE) and 32 from patients with systemic sclerosis were assessed. IgG and/or IgM anti-hsp73 antibodies were detected in 33% (19/57) of the patients with localized scleroderma. Among the three subtypes of localized scleroderma, generalized morphoea showed the highest incidence of anti-hsp73 antibodies (40%, 6/15). IgG and/or IgM anti-hsp73 antibodies were also detected in 9/30 samples (30%) from patients with SLE and in 13/32 samples (41%) from patients with systemic sclerosis, while the samples from the healthy controls were all negative for anti hsp73. By immunoblotting, specific binding of antibodies to hsp73 was confirmed with representative serum samples that were positive for anti-hsp73 in the ELISA. Our findings indicate that the presence of anti-hsp73 is an additional immunological abnormality in localized scleroderma. PMID- 7487147 TI - Photodynamic DNA-breaking activity of serum from patients with various photosensitivity dermatoses. AB - Various drugs and chemicals break the DNA strand under ultraviolet irradiation. This study aimed to clarify the DNA-breaking activity (DBA) of serum from 39 patients with various photosensitivity disorders and that from eight normal subjects. A mixture of serum and circular plasmid DNA was exposed to longwave ultraviolet radiation, and the photoinduced cleavage of plasmid DNA was examined by electrophoretic analysis. DBA was found in serum from patients with erythropoietic protoporphyria (2 of 2), drug-induced photosensitivity (3 of 5), chronic actinic dermatitis (1 of 12) and hydroa vacciniforme (1 of 1). DBA was not found in serum from patients with porphyria cutanea tarda, collagen diseases with photosensitivity, papulovesicular light eruption or pellagra. The inhibition profile of DBA by active oxygen scavengers was different between afloqualone- and tetracycline-induced photosensitivity and chronic actinic dermatitis. The present method was useful for the detection of serum phototoxicity and the investigation of the pathomechanisms of photosensitivity. PMID- 7487148 TI - The human hair follicle: glycoprotein-related antigenic profile of distinct keratinocyte populations in vivo and their alterations in vitro. AB - The cell surface expression of three glycoprotein antigens, as defined by the monoclonal antibodies BT 15, T 43, and MH 99, was investigated in follicular keratinocyte populations in vivo. In addition, the regulation of glycoprotein synthesis was studied in follicular and interfollicular keratinocytes cultured in vitro. The BT 15 antigen was strongly expressed in the inner root sheath and the area above Auber's line of the hair bulb, whereas the T43 antigen was mainly seen in the outer root sheath. Selectively high expression of the MH 99 antigen was found only in outgrowing germ buds of early anagen follicles. Radioimmunoprecipitation revealed strong signals with BT 15 in freshly prepared follicular keratinocytes, two to three times stronger than those in interfollicular keratinocytes, but the signals clearly decreased by 80% under continuing culture conditions. The T 43 antigen was found by FACS analysis and radioimmunoprecipitation in initially low amounts in both populations, but the signals increased dramatically (up to 50 times) in long-term cultures and in subcultures. The MH 99 antigen was also initially present only in low amounts, in interfollicular rather than in follicular keratinocytes, but its expression increased up to 15-fold with continuing culture and any differences between the two populations disappeared. Our investigation revealed that at least three populations of hair follicle keratinocytes are characterized by different surface glycoprotein antigens, clearly related to their state of differentiation and proliferation.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7487149 TI - Scanning electron microscopic observations of extracted terminal hair follicles of the adult human scalp and eyebrow with special references to the bulge area. AB - Scanning electron microscopic studies of human terminal hair follicles of the scalp and eyebrow have previously been limited to the hair shaft. In this study we investigated EDTA-treated extracted whole hair follicles in which most of the basal cell surface of the outer root sheath was well preserved. In the bulge area of scalp follicles there were many knob-like or villous projections. These were located in some specimens on one side of the follicle, while in others they were located around the entire circumference of the follicle. These projections were thought to represent the anchoring points of the branched follicular end of the arrector muscles. Ring-like elevations with groove-like depressions above and below were also observed surrounding the entire follicle. These were thought to represent the track of circumfollicular arrector muscles which depressed the outer root sheath when they repeatedly contracted. Most anagen eyebrow follicles showed morphological variations in the bulge area such as lattice-window-like structures and undulation patterns. In telogen follicles, the bulge became indistinguishable from the clubbed end. The lower end of these telogen follicles showed irregularly shaped bulge areas, but did not show lattice-window-like structures or undulation patterns as observed in anagen follicles. Interestingly, a hole was found in some bulge areas of both anagen and telogen follicles. Serial vertical sections of follicles revealed invaginated areas, which seemed to correspond to the openings seen in whole mounts. In vertical sections of eyebrow follicles some keratinocytes of the outer root sheaths of the bulge area were seen to be melanized to various extents. This phenomen was independent of hair cycle phase. PMID- 7487150 TI - Detection of Epstein-Barr virus DNA in anal scrapings from HIV-positive homosexual men. AB - Epstein-Barr virus (EBV) can infect epithelial cells as well as B lymphocytes. Infection of the male and female genital tracts has recently been demonstrated, and it has been suggested that the virus may be sexually transmissible. In our study we investigated whether EBV can be found in the anal region of sexually active homosexual men. Anal scrapings from HIV-positive homosexual men and a heterosexual control population were investigated using the polymerase chain reaction (PCR) to screen for EBV DNA. EBV DNA was detected in 8 of 27 anal samples (29.6%) from the homosexual men and 3 of 34 samples (8.8%) from the heterosexual men. Our study shows that, like the genital tract, the anal region can harbour EBV subclinically. This finding suggests that the anal region may be a reservoir for EBV and that sexual transmission of this virus may be possible. PMID- 7487151 TI - Detection of human papillomavirus DNA in nongenital seborrhoeic keratoses. AB - The histological similarities of seborrhoeic keratoses and common warts led to the investigation of the possible occurrence of human papillomavirus DNA (HPV DNA) in a large number of nongenital seborrhoeic keratoses using the in situ hybridization technique. All specimens derived from normal skin (n = 173) were negative for the applied HPV-DNA probe, whereas the HPV genome was detected in 34 of 173 seborrhoeic keratosis specimens (19.65%). Of 34 HPV-positive specimens, 15 contained types 6/11 and 14 types 31/33/35, and 5 showed no positive reaction to the applied types. These results suggest that a considerable percentage of nongenital seborrhoeic keratoses may be related to an HPV infection. PMID- 7487152 TI - Cutaneous warts: clinical, histological and virological correlations. PMID- 7487155 TI - Gas phase infrared spectra of 209 polychlorinated biphenyl congeners using gas chromatography with Fourier transform infrared detection: internal standardization with a 13C-labelled congener. AB - The gas phase infrared spectra of biphenyl and 209 polychlorinated biphenyls (PCB) are observed; 30 are presented in graphical form so that their uniqueness can be appreciated. The spectra were observed with a gas chromatograph linked via a light pipe with a Fourier transform infrared spectrometer. Unequivocal library matches (with match factors > 990 in 1000) can be performed with 50 ng of a congener injected into the chromatograph. Quantitation with the technique was briefly investigated with 2,2',5,5'-tetrachlorobiphenyl (BZ no. 52). Selective wavelength chromatograms of the most intense absorption (1457cm-1) were integrated. Septuplicate injections showed that 20 ng of the compound could be determined with +/- 4% precision. The resolution of the spectrometer was sufficient to distinguish between the 13C-labelled trichlorobiphenyl and the unlabelled material, indicating the possibility of internal or surrogate standardization of PCB analysis. PMID- 7487153 TI - Organochlorines in Pleuronectidae: comparison between three tissues of three species inhabiting the Northwest Atlantic. AB - Muscle, liver, and gonad from three species of flatfish, turbot, American plaice, and yellowtail flounder collected in the Northwest Atlantic were analyzed for a variety of organochlorine contaminants. Female flounder and plaice had lower concentrations of organochlorines than males, in all tissues. Comparing values to those previously obtained for Atlantic cod showed that concentrations in liver, the organ most often used in biomarker studies, followed the order: cod > turbot > plaice > flounder and correlated with lipid content. Muscle concentrations, which are important from a consumers perspective, followed the order: turbot > plaice > flounder > cod and were also correlated with lipids. The concentration of contaminants in gonads, the organ that informs on the future of a species were nearly undetectable. Cluster analyses demonstrated that the environmental behaviour of these contaminants can be subdivided into groups, consistent with their known chemical origin. Of the series of total and specific polychlorinated dibenzo-p-dioxins and furans studied, only tetrachlorodibenzofuran was present in most tissues. PMID- 7487154 TI - Contaminants of fishes from Great Lakes-influenced sections and above dams of three Michigan rivers: III. Implications for health of bald eagles. AB - Recently, there have been discussions of the relative merits of passage of fishes around hydroelectric dams on three rivers (Au Sable, Manistee, and Muskegon) in Michigan. A hazard assessment was conducted to determine the potential for adverse effects on bald eagles that could consume such fishes from above and below dams on the three primary rivers. The hazard assessments were verified by comparing the reproductive productivities of eagles nesting in areas where they ate primarily fish from either above or below dams on the three primary rivers, as well as on two additional rivers in Michigan, the Menominee and Thunder Bay. Concentrations of organochlorine insecticides (OCI), polychlorinated biphenyls (total PCBs), 2,3,7,8-tetrachlorodibenzo-p-dioxin equivalents (TCDD-EQ), and total mercury (Hg) were measured in composite samples of fishes from above and below hydroelectric dams on the Manistee and Muskegon Rivers, which flow into Lake Michigan, and the Au Sable River, which flows into Lake Huron. Mean concentrations of OCI, total PCBs, and TCDD-EQ were all greater in fishes from below the dams than in those from above. The hazard assessment indicated that current concentrations of Hg and OCI other than DDT (DDT+DDE+DDD) in fish from neither above nor below dams would present a significant hazard to bald eagles (Haliaeetus leucocephalus). Both total PCBs and TCDD-EQ in fishes from below the dams currently present a significant hazard to bald eagles, since their mean hazard quotients (HQ) were all greater than one. PMID- 7487156 TI - Polychlorinated biphenyls and 2,3,7,8-tetrachlorodibenzo-p-dioxin equivalents in eggs of double-crested cormorants from a colony near Green Bay, Wisconsin, USA. AB - Great Lakes colonial waterbirds have experienced poor reproduction and a greater incidence of birth defects than those in remote areas. An egg was collected from each of 1,000 marked cormorant nests at Spider Island (Lake Michigan). Nine pools comprised of three eggs were randomly selected for instrumental quantification of polychlorinated biphenyls (PCB) congeners, calculation of 2,3,7,8 tetrachlorodibenzo-p-dioxin equivalents (TEq) and measurement of equivalents by bioassay (TCDD-EQ). PCB analysis of the nine samples was semi-automated with high performance liquid chromatography (HPLC) columns including a porous graphitic carbon column. TEqs were calculated from concentrations of PCB congeners and bioassay-derived toxic equivalency factors (TEfs), and TCDD-EQ were measured directly with an H4IIE bioassay. Total PCB concentrations ranged from 9.7 to 38 micrograms/g, wet weight (ww). Mean concentrations of PCB 77, 126, and 169 were 2, 7, and 1 ng/g, ww. The mean TEqs and TCDD-EQ were 150 and 350 pg/g, ww, respectively. Thus, PCB congeners contributed less than 50% of the total TCDD-EQs as measured by the bioassay. PMID- 7487158 TI - Influence of salinity on copper and azide toxicity to larval topsmelt Atherinops affinis (Ayres). AB - Performance of a 7-d growth and survival toxicity test protocol using larval topsmelt, Atherinops affinis (Ayres), was evaluated for copper chloride and sodium azide at representative estuarine salinities. Results showed that topsmelt are amenable to toxicity testing at estuarine salinities ranging from 5 to 34/1000 since control survival was 100% in all toxicity tests. Sensitivity to both toxicants increased at lower salinities, with the LC50s for copper ranging from 205 micrograms/L at 34/1000 to 44 micrograms/L at 10/1000, and those for sodium azide ranging from 54 mg/L at 34/1000 to 7 mg/L at 5/1000. Larval tissue osmolality decreased with increasing copper concentration relative to control fish. Copper uptake was not affected by changes in salinity. This suggests that increased sensitivity to copper was due, in part, to the increasing physiological challenge of osmoregulation. It is also possible that cupric ion concentration increased at lower salinities, resulting in reduced larval survival. It is hypothesized that increased sensitivity to azide at lower salinity was due to the interaction between azide toxicity and increasing osmotic challenge. A second experiment with azide showed that larval acclimation time did not affect the interaction between salinity and azide toxicity. PMID- 7487157 TI - Environmental polychlorinated biphenyls: acute toxicity of landfill soil extract to female prepubertal rats. AB - Subsurface soil from a National Priorities List landfill containing about 2.5% polychlorinated biphenyls (PCBs) was extracted and the extract cleaned by Florisil slurry and alumina column chromatography. The refined extract contained 48 mg/mL PCB, mainly trichlorobiphenyls and tetrachlorobiphenyls, traces of polychlorinated naphthalenes, 125 micrograms/mL 2,2-bis-p-chlorophenyl-1,1 dichloroethylene (DDE), and low levels of chlorinated dibenzofurans. The refined extract was dissolved in corn oil and administered intraperitoneally to weanling (day 20) female rats on days 20 and 21; rats were terminated on day 22. Limited data indicated possible hematopoietic effects, including neutrophilia. There were no changes in relative uterus, kidney, or adrenal gland weights between total doses of 3 to 96 mg/kg total PCB. Relative liver weights increased significantly at 36 mg/kg and activities of P450s 1A1 (as ethoxyresorufin O-dealkylase) and 2B (as pentoxyresorufin O-dealkylase) increased at 12 mg/kg and plateaued at 36 (P450 1A1) or 48 (P450 2b) mg/kg. Serum total thyroxine (T4) declined significantly at doses of 36 mg/kg and greater; thyroid follicular epithelial cells were significantly larger within the same dose range, but the follicular colloid area decreased to less than 60% control values at 12 mg/kg and remained at this size through 72 mg/kg. Maximum mobilization of T4 apparently occurred at 12 mg/kg and attenuated measured declines in circulating levels. Even though a large proportion of proven and probable estrogenic chlorobiphenyls (CBs) were present, the lower amounts of more potent antiestrogenic aryl hydrocarbon (Ah) receptor agonists and/or decreased responsiveness because of low serum T4 levels may have antagonized the uterotropic response. PMID- 7487159 TI - The effect of emissions on heavy metals concentrations in cattle from the area of an industrial plant in Slovakia. AB - Attention was devoted to the effects of risk element-containing industrial emissions on the biological material of cattle bred on farms in the vicinity of a metallurgical plant in East Slovakia. The observations lasted two years and included four agricultural farms in the polluted area and one control farm outside this area. In 1991, 20, 30, 30, 30, and 30 samples of milk, muscle, liver, heart, and kidney tissue were analyzed, whereas in 1992, 30, 48, 48, 48, and 48 respective samples were taken. For Cu, Zn, Mn, Cd, Pb, and Ni analyses, the AAS method was employed, using the UNICAM 939 device, while Hg residues were determined on the TMA 254 analyzer. In 1991, of a total of 120 samples of tissue and organs, 12.5%, 9.16% and 0.8% were found to contain above-limit levels of Cd, Ni, and Cu, respectively. The concentrations of the remaining elements, viz., Zn, Mn, Pb, and Hg, did not surpass the permitted maxima. In 1992, 1.03%, 2.60%, 0.51%, and 5.20% of 192 samples of tissue and organs contained above-limit Cd, Cu, Ni, and Zn levels. PMID- 7487161 TI - The decline of mink in Georgia, North Carolina, and South Carolina: the role of contaminants. AB - Since the 1960s, mink (Mustela vision) populations in Georgia, North Carolina, and South Carolina have declined, especially in the coastal plain. A prior study suggested that the decline may stem from environmental contaminants. Based on water quality data from each state, we identified 17 substances potentially related to the decline: aldrin, dieldrin, endrin, DDD, DDE, DDT, PCBs, chlordane, alpha-BHC, toxaphene, dibenzofuran, copper, chromium, cadmium, lead, arsenic, and mercury. Mink livers were analyzed for PCB and organochlorine pesticides, and kidneys and femurs were analyzed for metals. Reference sample concentrations from piedmont, mountain, and foothill locations were compared to state coastal plain totals and counties. PCBs for Georgia, dieldrin for South Carolina, and endrin and aldrin for North Carolina were significantly higher than the piedmont reference group. Liver PCB concentrations were higher than those known to cause mink reproductive dysfunction. Mercury concentrations were significantly higher in coastal plain mink from all three states and were in the range of those known to cause impacts to reproduction, growth, and behavior to wild mink. It is unknown what concentrations of cyclodienes cause reduced reproduction or other physiological effects in mink, but the levels reported here probably indicate background concentrations that do not contribute to the decline. PMID- 7487163 TI - Jack of all trades and master of one. PMID- 7487162 TI - Protective effects of free radical scavengers and antioxidants against smokeless tobacco extract (STE)-induced oxidative stress in macrophage J774A.1 cell cultures. AB - Previous studies have demonstrated that an aqueous smokeless tobacco extract (STE) administered in an acute oral dose to rats induces an enhanced induction of hepatic mitochondrial and microsomal lipid peroxidation, hepatic nuclear DNA single strand breaks, enhanced excretion of urinary lipid metabolites, including malondialdehyde, formaldehyde, acetaldehyde and acetone, and increased production of nitric oxide (NO) by peritoneal macrophage cells. These observations indicate that STE induces the production of oxygen free radicals. We have therefore examined the in vitro incubation of cultured J774A.1 macrophage cells with STE on the release of the enzyme lactate dehydrogenase (LDH) into the media as an indicator of cellular membrane damage and cytotoxicity. The amount of LDH released by STE was both concentration- and time-dependent. The cytotoxicity of STE to macrophage J774A.1 cells in culture was further determined from percent viability after various periods of incubation. The addition of 250 micrograms STE/ml to the cultured J774A.1 cells resulted in a 2.9-fold increase in the release of LDH. Individual coincubation with superoxide dismutase (SOD), catalase, mannitol, and allopurinol had no significant effect on the release of LDH into the culture medium, while a combination of the four free radical scavengers resulted in a 59% decrease in the STE-induced release of LDH. At 75 microM concentrations of viramine E and vitamin E succinate, approximately 28% and 41% inhibitions were observed in STE-induced LDH leakage, respectively. Taken together with previous studies, the results indicate that STE activates macrophage cells, resulting in the production of reactive oxygen species.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7487164 TI - Special report: protection and advocacy: what nurses need to know. AB - There is a national system of protection and advocacy programs for people with mental illness, developmental disabilities, and other disabilities. These programs investigate allegations of abuse and neglect and pursue legal, administrative, and other appropriate remedies to protect the rights of individuals with disabilities. Nurses have a critical role in advocating and protecting rights of consumers with psychiatric disabilities and identifying and reporting incidents of abuse and neglect. The unique role of the nurse within practice, research and education regarding protection and advocacy is discussed. PMID- 7487160 TI - Dietary exposure of mink to carp from Saginaw Bay, Michigan: 2. Hematology and liver pathology. AB - The effects of consumption of environmental contaminants contained in carp (Cyprinus carpio) from Saginaw Bay, Michigan on various hematological parameters and liver integrity of adult female mink (Mustela vision) were determined. Mink were fed diets that contained 0 (control), 10, 20, or 40% carp prior to and throughout the reproductive period (182 days). The diets contained 0.015, 0.72, 1.53, and 2.56 mg polychlorinated biphenyls (PCBs)/kg diet and 1.0, 19, 40, and 81 pg 2,3,7,8-tetrachlorodibenzo-p-dioxin equivalents (TEQs)/g diet, respectively. Mink fed the diets containing carp showed a general dose-dependent occurrence of clinical signs commonly associated with chlorinated hydrocarbon toxicity, including listlessness, nervousness when approached, anorexia, and melena. Erythrocyte counts were less in mink exposed to Saginaw Bay carp than in controls, while the number of white blood cells was greater than in controls. Significant differences (p < 0.05) in the concentrations of neutrophils, lymphocytes, monocytes, and eosinophils were also found between the control and carp-fed groups, but are considered to be of limited clinical or biological importance. Hematocrit values for the mink fed the 20 and 40% carp diets were significantly less than those of mink in the control and 10% carp groups. There were no significant differences in hemoglobin concentrations among the groups. Necropsies revealed enlarged yellowish livers in many of the carp-fed mink, especially those fed the 40% carp diet. Liver, spleen, and lung weights of carp fed mink were significantly greater than those of control mink.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7487165 TI - Clinical outcomes and satisfaction of patients of clinical nurse specialists in psychiatric-mental health nursing. AB - Survey research was conducted to examine clinical outcomes and satisfaction of patients of psychiatric mental health clinical nurse specialists (CNSs). Patients who had terminated from outpatient psychotherapy with 6 CNSs in 1993 were mailed a questionnaire (N = 223). Follow-ups by mail yielded a response rate of 45% (n = 100). The questionnaires included the Profile of Mood States-Short Form ([POMS SF]; McNair, Lorr, & Droppleman, 1992). Quality of Life Function ([QOL]; Lehman, 1991), and Patient Satisfaction Scale (Baradell, 1994). Paired difference t-tests were used to evaluate clinical outcomes. Percentages were used to report satisfaction, and Pearson correlations were used to examine the relationship between clinical outcomes and patient satisfaction. The mean age for respondents was 37 years; 82% were female. Diagnoses included depression (46%), adjustment disorders (34%), anxiety (10%), and other (10%). Patients reported significant improvement in all clinical symptoms: anxiety, depression, anger, confusion, fatigue and vigor. Patients reported significant improvement in all domains of QOL: family, social, and job. Patients reported a very high level of satisfaction with the care provided. The more clinical improvement the patients reported, the more satisfied they were with the care provided. If nurses are to be included in a reformed health care delivery system in the future, additional research is essential. PMID- 7487166 TI - Agitation: measurement, management, and intervention research. AB - Strategies for controlling and managing agitation of patients in long-term care populations has been identified as a research priority. Unfortunately, there is a paucity of literature on empirically tested therapeutic interventions for agitation. This manuscript summarizes the literature related to the measurement, management, and interventions for agitation and identifies a nursing research agenda for conducting research in this area. Nonpharmacological and nonrestraint interventions are highlighted and categorized by (1) alterations in care given, (2) environmental adaptations, and (3) behavioral approaches. Future research areas identified by the authors focus on additional types of environmental, interpersonal, and behavioral interventions as well as theoretical predictors of agitation; the influence and role of nursing personnel and caregiver; and the legal and ethical issues involved in caring for patients who manifest agitation. PMID- 7487167 TI - The development of the self-care ability to detect early signs of relapse among individuals who have schizophrenia. AB - A qualitative study using interpretive interactionism investigated how individuals with schizophrenia develop an ability to detect early signs of relapse. A purposive sample of 15 respondents were interviewed about their illness experiences. Three sequential processes were identified. Responding reactively to psychic pain, initiated by a desire to escape from overwhelming distress, resulted in the differentiation of tolerable and intolerable pain. Refining perceptions of psychic pain followed and resulted in the identification of specific thoughts, feelings and behaviors associated with different levels of distress. Finally, actively reducing psychic pain resulted in a proactive surveillance for specific signs of increasing distress. PMID- 7487168 TI - Symptoms of depression among female nursing students. AB - This study examines the extent to which depressive symptoms in female nursing students are affected by specific stressors and coping styles. Three hypotheses were examined for differences in symptoms of depression scores and a model was tested for predicting depression in female nursing students. Responses were gathered from three questionnaires (Hassles and Uplifts Scales, Symptoms of Stress Inventory, and Coping Styles) from 408 female baccalaureate, master's and doctoral students from a major Midwest research university. Symptoms of depression were not significantly different among the students and were as high as a comparative group of stress management clients. Path analysis was used to examine the patterns of stressors (hassles, uplifts, personal and student-related stress), coping (coping styles, habits, quitting school, and drug use), and a biological factor (depression around the menstrual cycle) that predicted symptoms of depression. All hypothesized variables had direct paths to symptoms of depression; the path model explained 58% of the variance. Results from this study support the biological and psychological theories of depression in women and raise important questions of particular relevance to women, nursing students and educators. PMID- 7487169 TI - Communicating with persons with Alzheimer's disease: experiences of family and formal caregivers. AB - The purpose of this exploratory study was to examine and compare communication processes used by family members and formal caregivers to manage behavioral problems such as fearfulness, agitation, and wandering, that commonly occur when caring for persons with Alzheimer's Disease (AD). Two groups of caregivers of persons with AD (formal and family) comprised the sample for the study. Two themes emerged from the focus group interviews: environmental adjustments and reassurance. Results suggest that interventions must be individualized. Enhancing the caregiver's skills to manage disruptive behavior may prolong his or her ability to provide in-home care for the family member with AD. PMID- 7487170 TI - Improving nurses' responses toward substance-misusing patients: a clinical evaluation project. AB - Practitioners in acute and primary care settings often fail to identify and intervene with chemically dependent patients. Counterproductive attitudes, lack of knowledge, and poor clinical skills impede early identification, treatment, and referral. In response to these problems, an educational intervention was designed to improve practicing nurses' recognition of and responses to substance misusing patients. Results of the pretest posttest comparison group evaluation suggest that educational interventions were influential in improving nurses' confidence in caring for substance misusing patients. Although data reflecting changes in attitudes were less compelling, the improvement in treatment optimism is a positive change that has important clinical implications. PMID- 7487172 TI - [Incidental urologic neoplasm: towards a new management approach]. PMID- 7487171 TI - Patient perceptions of self-induced water intoxication. AB - This study comprehensively examines self-induced water intoxication (SIWI) from a patient perspective including demographics, reasons for seeking fluids, patterns/behaviors of fluid seeking, and symptoms frequently experienced while in a state of SIWI. The subjects were 45 of an original convenience sample of 62 individuals with a serious mental illness (SMI), hospitalized in a long-term state psychiatric facility, who engaged in self-induced water intoxication. All participants were interviewed in a structured format to complete a 40-item Likert type questionnaire developed for the study, titled the Self-induced Water Intoxication Questionnaire (SIWIQ). In the study, the majority of participants were smokers, and reported no past problem with alcohol. SIWI occurred more in males than females, and was more predominant in those participants who had longer hospital stays. Anger and vomiting were found to be the two most predominant symptoms experienced when excess fluid consumption occurred. Behaviors of drinking from the shower, the toilet, and one's own urine are consistent with findings of previous studies and illustrate the difficulty in keeping individuals with SIWI from fluids. Data show that participants with SIWI experience considerable anxiety and cognitive difficulties and express these as reasons for engaging in excess fluid consumption. Boredom, obtaining a high, and sad mood were also predominant reasons identified for excess fluid drinking. Significant relationships were found and discussed. The findings provide support for the position that SIWI represents an attempt at treatment by the dysfunctional individual and is pursued for anxiolytic effects and alleviation of boredom. The data support the idea of approaching the problem from a dysfunctional coping framework, realizing that SIWI is a very complex problem, needing examination and intervention at multiple levels, beyond exclusive focus on fluid control. PMID- 7487173 TI - [Potential of ultrasonography in urologic diagnostic algorithms]. AB - OBJECTIVES: The present study analyzed the place of ultrasound in the urological diagnostic protocols. Currently, most of the patients undergo some type of ultrasonographic evaluation at the outset or at some time during follow up. Our experience concerning the results, limitations and errors of this imaging technique are presented. METHODS: We reviewed the indications for US in different pathologies and its use together with other diagnostic techniques. Simple, cost efficient algorithms are described. RESULTS: Its excellent performance permits making a correct diagnosis in many patients and remarkably simplifies evaluation in others. Its accuracy obviates the need for other explorations in certain pathologies. CONCLUSIONS: Ultrasound must be included in the urological armamentarium and should be considered as an extension of physical examination. Its accuracy and efficacy have been demonstrated. Furthermore, it is simple and easy to use, low-cost, and remarkably simplifies our diagnostic algorithms. PMID- 7487174 TI - [Urethroplasty with lyophilized human dura mater]. AB - OBJECTIVE: This article reports our experience with lyophilized human dura in the treatment of complex urethral stenosis. METHOD: Seven patients with iatrogenic/inflammatory urethral stricture underwent a 13 cms long x 1.5 cms wide medial urethral augmentation. RESULTS: During a mean follow-up of 23 months, good results accounted for 85.7%. CONCLUSION: Lyophilized dura patches constitute a suitable choice in selected patients who, for local or systemic reasons, are not good candidates for other urethroplasty procedures. PMID- 7487175 TI - [Survival of patients with upper urothelial tumor]. AB - OBJECTIVE: A descriptive analysis of 35 patients with transitional cell carcinoma of the upper urinary tract is presented. The actuarial survival was analyzed to determine which circumstances associated with superficial bladder cancer had a prognostic significance. METHODS: The records of 35 patients with upper urothelial tumor diagnosed from 1987 to 1993 were retrospectively analyzed. A descriptive statistical analysis was performed on all the variables. Correlation was determined by Pearson's Xi2 test, Yates' and Fisher's exact test. McNemar's test was used to determine which diagnostic method had the highest yield. The actuarial survival data were analyzed with the Kaplan-Meier and Mantel-Haenzel test. RESULTS: Patient mean age was 67.74 yrs, mean follow up was 26.83 months, 40% were ureteral tumors, 25.7% were pelvic tumors, 60.6% were superficial tumors and 62.6% were GI and GII, 48.5% were associated with bladder tumors and no correlation was found between the grade and stage of bladder relapses and the upper urinary tract tumors. Intravenous urography demonstrated function was abolished in 45.5%. Cytology gave the highest diagnostic yield (84.6%). Nephroureterectomy with perimeatal cystectomy was the surgical technique most frequently used (57.1%). To date, 71.4% of the patients have survived. Significantly women demonstrated the most undifferentiated cellular grade (p < 0.001), although no prognostic value was found for sex. 95% of the patients with non-infiltrating tumor stages and 95% of the patients with GI-GII tumors survived, whereas 61.5% of the patients with infiltrating tumors and 58.3% of those with GIII tumors did not survive (p < 0.002). Evidently, the grade of cellular differentiation and the stage of infiltration were interrelated (p < 0.002). Analysis of the actuarial survival data showed that the probability of survival after 24 months (mean follow up) was 67.07% and the probability of reaching 74 months (maximum follow up) was 60.04%. CONCLUSIONS: After a comparative analysis of the survival with the Mantel-Haenzel test, it was found that the grade of cellular differentiation (p < 0.001) and stage of infiltration (p < 0.01) had significant differences in the survival curves. PMID- 7487176 TI - [Assessment of arterial hypertension associated with renal transplantation using Doppler ultrasonography]. AB - OBJECTIVES: The incidence of arterial hypertension post-renal transplantation has been reported to be 48.6%. The present study investigated the usefulness of echo Doppler in detecting renal artery stenosis and the effects of arterial pressure control and hypotensive agents on the vascular resistance of the renal graft. METHODS: A total of 234 echo Doppler studies were done in 110 patients. The pulsatility index (PI), resistance index (RI), acceleration and mean velocity (MV), maximum systolic (SVMX) and minimum diastolic (DVMN) velocities of the external iliac, renal, segmental, interlobar and arcuate arteries were determined. RESULTS: In the stenotic segment, an elevation of SVMX was observed, followed by turbulent flow in the post-stenotic segment in 73% of the cases with arterial stenosis. The hypertensive patients had a higher external iliac artery RI than the normotensives. The beta-blockers reduced the RI in large vessels (external iliac), the vasodilators in medium-sized extraparenchymal vessels (segmental) and the calcium antagonists changed the PI in the smaller caliber intraparenchymal vessels (interlobar and arcuate). CONCLUSIONS: Evaluation by echo Doppler is the first diagnostic approach in patients suspected as having arterial stenosis of the renal graft. It permits evaluating blood flow changes from hypertension and hypotensive agents. PMID- 7487177 TI - [Effects of urethropexy on urination. A price to pay?]. AB - OBJECTIVES: To evaluate the voiding repercussions of urethropexy. METHODS: We carried out a clinical and urodynamic study in 70 patients subjected to different techniques of urethropexy (Kelly, Raz, Marshall-Marchetti-Krantz, Burch and simplified Ramirez) for stress urinary incontinence. We evaluated the voiding repercussions of urethropexy. RESULTS: There was a significant increase in the obstructive symptoms with urethropexy (29% of the patients) although there was no relationship with the variation of the urinary flow rate. It also significantly diminished the urinary flow rate (85% of the patients) and increased the postvoiding volume significantly (3.68 times the average before surgery). The increase in postvoiding volume occurred more frequently in patients with diminished urinary flow rate before urethropexy (60% of the cases). The Raz technique showed less urodynamic data of voiding disturbances. CONCLUSIONS: Urethropexy causes voiding disturbance. We believe the decrease in urinary flow rate is related to the increase in urethral resistance produced by urethropexy. In the cases with previously diminished urinary flow rate, reduction in bladder contractility is added to the former mechanism. PMID- 7487178 TI - [Obstruction of the lower urinary tract: contractile capacity of the detrusor and obstruction degree. Analysis of pressure/flow curves]. AB - OBJECTIVES: This study attempts to optimize information from pressure/flow curves of patients with obstructive symptoms. METHODS: We performed a computerized analysis of the pressure/flow ratio of 50 consecutive patients with benign prostatic hyperplasia and irritative and obstructive symptoms. All patients were evaluated urodynamically. This model (pressure/flow diagram) will allow application of this test (expressed as flow and pressure measurements) to urethral outflow resistance and detrusor power or energy. Outflow obstruction is classified into seven categories (O-VI) and detrusor contractility in four (very weak, weak, normal and strong). RESULTS: The suitability of urodynamic analysis was demonstrated. Those cases considered as indeterminate by the conventional method can be obviated by computerized analysis. The degree of obstruction and the contractile capacity were determined in these cases. CONCLUSIONS: This method provides more accurate information on pathophysiology of voiding, allowing for better diagnosis of outflow obstruction or poor detrusor contractility and avoids the diagnosis of "undetermined". PMID- 7487179 TI - [Which is the role of laparoscopic surgery in the treatment of cryptorchism?]. AB - OBJECTIVES: The present study analyzed the indications for laparoscopic surgery in cryptorchidism. METHODS: The clinical, surgical and anatomopathological data of 2000 cryptorchid testes submitted to surgery from 1972 to 1993 were reviewed. RESULTS: 17.75% (355 testes) were not palpable at physical examination; of these, 3.45% (69 cases) were anorchid and only 5.4% (108 testes) of the remaining 14.3% were true intra-abdominal testes. The other 8.9% were localized elsewhere. Scrotal descent was achieved only by funiculolysis in 68% of the cases. The remaining 28% (30 testes, since the technique used was not reported in 5) were brought to the scrotum using two-stage procedures, Stephen-Fowler and autotransplantation (one case), which could have been replaced or preceded by laparoscopy. Of the 30 testes, 10 were removed. CONCLUSIONS: Since only 5% (20 testes) can benefit from laparoscopic techniques and, moreover, 68% have a very low tubular fertility index and there is an increased possibility of progressing to malignancy, if other than standard surgical techniques are warranted in cases with unilateral cryptorchidism, testis removal is preferable, which further reduces the application of laparoscopy in cryptorchidism. PMID- 7487180 TI - [Endourologic treatment of uretero-ileal stenosis]. AB - OBJECTIVES: Ureterointestinal stricture represents a serious problem whose management has traditionally been by open surgery. Endourological treatment is an alternative that is generally free from the complications of open surgery, but with a lower success rate. The efficacy of endourological treatment is analyzed in 14 cases of ureteroileal stricture (12 patients) and the different treatment options are discussed. METHODS: Ureteroileal stricture was managed endourologically in 9 of 13 cases (69%) by dilatation (6 cases), incision+dilatation (2 cases) and stenting (1 case). RESULTS: Of the 6 cases submitted to dilatation, 3 recurred at 6 months, 2 remained patent at 18 and 24 months, respectively, and 1 was lost to follow up because the patient underwent reimplantation (40% success rate). The two cases submitted to incision are patent at 18 and 19 months, respectively. CONCLUSIONS: The overall success rate for endourological treatment of ureteroileal stricture is 28% (4/14 cases) at 19 months mean follow up. PMID- 7487181 TI - [Ureteral stenosis secondary to methacrylate used in total hip arthroplasty]. AB - OBJECTIVES: The urological complications in orthopedic surgery, the mechanisms of injury and its treatment are discussed. METHODS/RESULTS: Herein we describe a case of ureterohydronephrosis arising from extrinsic ureteral compression by cement utilized in total hip arthroplasty in a patient with a solitary kidney. CONCLUSIONS: The risk of acetabular perforation and subsequent migration of the cement into the pelvic cavity must be taken into account in patients undergoing hip replacement surgery. Conservative endourological management should first be attempted in the event of ureteral involvement, although resolution is by surgical repair in most cases. PMID- 7487182 TI - ["Sand-clock" lithiasis on prostatic transurethral resection bed]. AB - OBJECTIVES: Although uncommon, prostatic surgical bed lithiasis may present among post-TURP complications. The present study underscores the importance of diagnosis and treatment. METHODS/RESULTS: We report on a patient with magnesium ammonium phosphate hourglass lithiasis in the prostatic surgical bed one year and a half following transurethral resection due to adenomyomatous prostatic hyperplasia that was successfully treated by intracorporeal lithotripsy with the Lithoclast. The physiopathological, clinical, diagnostic and therapeutic aspects are discussed. CONCLUSIONS: This condition should be suspected in patients with dysuric syndrome post-TURP. The importance of simple complementary diagnostic procedures as DRE and plain abdominal films in discarding lithiasis in underscored. We consider retrograde endoscopic manipulation and ultrasonic lithotripsy or penumatic lithotripsy with the Lithoclast to be the safest procedures for this type of lithiasis. PMID- 7487183 TI - [Metachronous metastasis in ureteral stump secondary to ipsilateral renal adenocarcinoma]. AB - OBJECTIVES: Ureteral metastasis from renal adenocarcinoma is uncommon. The rarity of this disease prompted us to report on the present case, with special reference to diagnosis. METHODS/RESULTS: A patient with metachronous metastasis in the ureteral stump from clear cell renal adenocarcinoma resected three years earlier is described. The etiopathogenic, clinical, diagnostic and therapeutic aspects are briefly reviewed. CONCLUSIONS: Ureteral metastases, although uncommon, are generally ipsilateral to the renal tumor and arise from aggressive renal carcinomas that are locally advanced with vascular and/or lymphatic spread. We underscore the importance of retrograde pyelography and ureterorenoscopy in the diagnostic workup of all patients with gross hematuria following nephrectomy due to hypernephroma. PMID- 7487184 TI - [Skin metastasis from transitional cell carcinoma of the bladder]. AB - OBJECTIVES: An additional case of cutaneous metastasis arising from transitional cell carcinoma of the bladder is described. Its form of presentation, dissemination pattern, pathological features, clinical course and treatment are briefly discussed. METHODS: This article reports on a 62-year-old male with bladder carcinoma stage PT3B, PN0, PMx, who underwent radical cystoprostatectomy and detubularized ileal neobladder four months earlier. RESULTS: A solid tumor with an ulcerated surface was found on the dorsal side of the left shoulder. Microscopic analysis disclosed neoplastic cells resembling transitional cells in dermis, the first manifestation of distant tumoral spread. Patient survival was only two months. CONCLUSIONS: Transitional cell carcinoma of bladder metastasizing to the skin is an uncommon finding despite the high incidence of this tumor, although more cases are being reported. Skin metastasis generally presents in the late stages of this disease and indicates a poor outcome. PMID- 7487185 TI - [Inflammatory aorto-iliac aneurysm. A cause of extrinsic ureteral obstruction]. AB - OBJECTIVE: A ureteral pathology arising from inflammatory aneurysm of the abdominal aorta is described. METHODS/RESULTS: A case of inflammatory abdominal aortic aneurysm causing entrapment of both ureters, destruction of a renal unit and ureterohydronephrosis of the contralateral kidney is described. CONCLUSIONS: Treatment of inflammatory aneurysm of the abdominal aorta is by surgical repair with a prosthesis. Endourological urinary drainage is warranted prior to surgical repair, but ureterolysis to preserve renal function is unnecessary. PMID- 7487187 TI - Heparin and dialyzer membranes during hemodialysis: a literature review. AB - When patients with end stage renal disease (ESRD) are hemodialyzed, the thrombosis/hemorrhage dilemma occurs. The goal is to give enough, but not too much anticoagulant during the dialysis procedure. Heparin is the anticoagulant most widely used to prevent clotting while blood is in the dialysis circuit. This article discusses the relationship among blood, heparin and dialyzer membranes during the hemodialysis procedure. The information presented will help nephrology nurses recognize the necessity to reevaluate heparin prescriptions each time dialyzer changes are made or the medical conditions of patients change. PMID- 7487186 TI - Ureteral lithiasis: in situ piezoelectric versus in situ spark gap lithotripsy. A randomized study. AB - OBJECTIVES: In situ extracorporeal lithotripsy (EL) is the treatment of choice for ureteral lithiasis at our institution since the introduction of the painless lithotripters in clinical practice. The major objections to such clinical approach are: difficult ultrasound localization of ureteral calculi (if an ultrasound-guided lithotripter is used); lower energy of the painless piezoelectric compared to the spark gap system. To evaluate the efficacy of in situ EL of ureteral stones and to compare two different lithotripters (the spark gap fluoroscopically-guided Dornier HM3 versus the piezoelectric ultrasound guided Wolf Piezolith 2300), we conducted a prospective randomized study. METHODS: 70 patients were randomly divided into two groups according to the following inclusion criteria: radiopaque stones of lumbar or prevesical tract (iliac and pelvic stones were excluded); stones with largest diameter not more than 2 cm. RESULTS AND CONCLUSION: The evaluation of the treatment was done by plain film (KUB) at 24 and 72 hrs and by ultrasound at 48 hrs to assess stone fragmentation using migration of the fragments more distally into the ureter as criteria for success. The results were comparable: 76.6% for extracorporeal piezoelectric lithotripsy (EPL) and 74% for extracorporeal shock wave lithotripsy (ESWL). Better results were observed in both groups for the prevesical stones. The focussing time required was also comparable. PMID- 7487189 TI - A unique dialysis quality management experience: an interview with novelist James A. Michener. Interview by Alex Rosenblum. AB - As a renal quality management nurse, I am always looking for new and interesting ways to share information and comparative data to educate and motivate fellow renal professionals. The intent of the sharing of the information is to stimulate quality activity. As a quality improvement project this interview with novelist James Michener, a patient with end stage renal disease, provides ANNA Journal readers with an increased understanding of the importance of providing quality care to the lives of people we care for. Recently, Michener provided an excellent opportunity to share his own experience in dealing with renal failure and dialysis therapy. His celebrity status and openness in discussing many issues related to his renal failure furnished a unique quality improvement opportunity. PMID- 7487188 TI - Attitudes of dialysis patients and caregivers regarding advance directives. AB - OBJECTIVE: The objective of this study was to describe and compare the attitudes of dialysis patients, their relatives/friends, primary nephrologists, and nurses regarding advance directives. DESIGN: A descriptive comparative design was used. SAMPLE/SETTING: A total of 50 dialysis patients from a large teaching hospital in western Canada participated in the study. For every patient who responded, a relative/friend, physician, and nurse completed the questionnaire. METHODS: The questionnaires were designed for the purposes of the study. These questionnaires were mailed to dialysis patients, who were asked to identify a relative/friend to complete a similar questionnaire. Nurses and physicians also completed questionnaires. RESULTS: Overall, 84% of patients indicated that they believed that it was important to leave a living will, but only 18% of these patients had actually left an advance directive. However, 52% of patients and relatives/friends believed that major treatment decisions should be left to the physician, a perception that was not shared by physicians and nurses. CONCLUSIONS: It appears that patients, family/friends, nurses, and physicians in this study had differing perceptions regarding advance directives. It is important that health care professionals realize that these differences may exist and that further research should address strategies to increase our understanding of patient decisions in this area. PMID- 7487190 TI - Developing an epoetin alfa medication protocol to improve patient care and foster collaboration. AB - This article describes the process undertaken by the nursing staff, management staff, and education department of a community hospital-based outpatient dialysis unit to address the numerous issues related to the management of anemia in their dialysis population. Through good observations and assessments made by the nursing staff, major problems related to anemia management were identified. As a result, staff members developed a medication protocol and a tracking tool to better manage the anemia associated with chronic renal failure. Evaluation of this protocol and tracking tool demonstrated improvement in two clinical patient outcomes and two unit management outcomes. PMID- 7487191 TI - Addition to the 1995 Educational Resource Directory. PMID- 7487192 TI - Successful hemodialysis in the elderly. PMID- 7487193 TI - CAPD catheter exit site healing and clean dressing techniques. PMID- 7487194 TI - A review of heparin use in hemodialysis and peritoneal dialysis. PMID- 7487195 TI - Epoetin alfa--focus on the geriatric patient. Case study of the anemic patient. AB - According to the Health Care Financing Administration, the largest group receiving dialysis therapy in the United States, constituting 41%, is persons age 65 and older. With 25% of the population expected to be in this age group by the middle of the next century, increasing attention must be paid to this developing "majority patient population." This article addresses the special requirements, major challenges, and management strategies needed to treat older patients with end-stage renal disease. PMID- 7487196 TI - Leadership positions require leaders, not managers. PMID- 7487197 TI - Community survey of diarrhoea in children under 5 years in Kinshasa, Zaire. AB - This community based survey was undertaken to assess the prevalence, characteristics and risk factors of diarrhoea in children < 5 years of age in an urban zone, at Kinshasa, Zaire. 155 community cases selected by cluster sampling, 155 age-matched controls, and 18 children with diarrhoea seen at a health centre (HC) were examined. The diarrhoea prevalence rate was 6.5%. The highest risk of persistent diarrhoea was in children of 2 to 3 years, non breast-fed, with more than one enteric agent in their stools and living in households without electricity. The rates of detection of Strongyloides stercoralis, Entamoeba histolytica and Salmonella, and heavy infections of Trichuris trichiura and Trichomonas hominis were significantly higher in cases (p < 0.05). The rate of detection of Cryptosporidium was rather high, specially in HC cases (22.2%). It was also found in both diarrhoeal (14.8%) and non-diarrhoeal specimens (12.9%). There was a lack of association between the presence of faecal white blood cells and enteric bacteria, and also between the presence of faecal red blood cells and E. histolytica, which might be due to the frequent practice of rectal injections and suppositories. A mother's perception of fever and stool aspect was fairly in agreement (respectively 70% and 53%) with that of the investigators. The high rate of dehydrated children (50.9%) may be due to the dehydration definition applied at the HC. The survey's results were used to improve the local case management flow chart and to adjust educational activities directed at mothers. PMID- 7487198 TI - [Surveillance of the nutritional status of the population in Kinshasa, Zaire (1991-1994)]. AB - Six anthropometric surveys using a random cluster sampling design have been organised in the capital of Zaire between November 1991 and March 1994. The prevalence of acute malnutrition fluctuated with the seasons and followed the cyclic movements of food prices on the market. The general tendency was rising. In March '94, 10.7% (95% confidence interval: 8.8-12.9%) of the children was suffering of acute malnutrition. Nevertheless, the standardised prevalence remained stable. This may suggest that the most disadvantaged living at certain poor zones of the city disconnected, while the rest of the population could still manage to maintain their nutritional status. The hypothesis of increasing heterogeneity is supported by the raising of the design-effect. PMID- 7487199 TI - Non-response to a recombinant pre-S2-containing hepatitis B vaccine: association with the HLA-system. AB - In this study, the immunogenicity of a recombinant pre-S2 containing hepatitis B vaccine was determined in a cohort of 147 homosexual men. Both seroconversion rate and geometric mean titers indicated that pre-S2 was less immunogenic than HBsAg. Forty-eight subjects, including nine non-responders and seven high responders to HBsAg, were tested for HLA Class I and II antigens to study a possible association between HLA-type and non-response. Both HLA-B8 and HLA-DR3 were clearly overrepresented in non-responders to HBsAg. Non-response to pre-S2, observed in 7/9 non responders and 6/39 responders to HBsAg, was not associated with any of the tested HLA markers. PMID- 7487200 TI - Study of HTLV-I antibodies in CSF and serum of neurolathyrism patients in Bangladesh. AB - Neurolathyrism is a form of human spastic paraparesis related to the overconsumption of the legume Lathyrus sativus or grass pea (Khesari in Bangladesh) containing the neurotoxin 3-N-oxalyl-2,3-diaminopropanoic acid (beta ODAP). The clinical symptoms of neurolathyrism are similar to those of Tropical Spastic Paraparesis. In order to eliminate the proposed causative agent of TSP (HTLV-I) as a potential cause of the symptoms ascribed to neurolathyrism, a total of 444 diagnosed lathyrism patients were screened for HTLV-I antibodies. 50 CSF and 394 serum samples were collected from male (415) and female (29) patients. Only 4 serum samples were found sero-positive for HTLV-I. This agrees with the assumption that overconsumption of beta-ODAP containing Lathyrus seeds, and not HTLV infection, is the causative agent for neurolathyrism. PMID- 7487201 TI - Rift Valley fever epizootic in small ruminants in southern Mauritania (October 1993): risk of extensive outbreaks. AB - Following the 1987 Rift Valley fever (RVF) epizootic/epidemic in southern Mauritania, RVF antibody prevalence in small ruminants decreased significantly from 1988 to 1992. In October 1993, at the end of the rainy season, an active RVF virus transmission was detected in several locations of southern Mauritania in small ruminants, in association with an increase of abortions. RVF IgM and IgG antibodies were recorded respectively in 65/390 (16.67%) and 73/390 (18.72%) animals. RVF IgM antibody prevalence was significantly higher in the > or = 2 years old group (26.02%) than in the 0.5- < 2 year old group (7.20%) (p = 0.01). The enzootic maintenance of RVF virus is discussed, as well as the risks of extension by traditional nomadic movements of herds. PMID- 7487202 TI - Efficacy and tolerance in adults of a short (3 days) course of quinine for uncomplicated falciparum malaria. PMID- 7487203 TI - Evaluation of sodium dodecyl sulfate (SDS) as a haemolytic agent for the detection of microfilariae and trypanosomes in the blood of cattle. PMID- 7487205 TI - Some operational aspects of the use of personal protection methods against malaria at individual and community level. PMID- 7487204 TI - [In memoriam Professor emeritus Dr. Pieter Hendrik van Thiel. 13 April 1895 - 23 June 1993]. PMID- 7487206 TI - Preoperative chemotherapy: where do we go from here? PMID- 7487207 TI - Conservation surgery after primary chemotherapy in large carcinomas of the breast. AB - OBJECTIVE: The authors evaluated the utility of preoperative chemotherapy in patients with large size breast carcinoma, with a view to rendering a conservative surgical approach possible or easier. SUMMARY BACKGROUND DATA: Two hundred twenty-six of 227 patients with breast cancer involving a tumor larger than 3 cm at greatest dimension were candidates for mastectomy. They were treated with various primary preoperative chemotherapies and evaluated for surgery. METHODS: After administering various chemotherapeutic regimens, the authors reevaluated the patients' conditions clinically and radiologically to plan definitive surgical treatment. If the tumor diameter was sufficiently reduced, quadrantectomy was planned; otherwise, mastectomy was performed. Complete axillary lymph node dissection was done in all cases. RESULTS: In 90% of the cases, the size reduction was sufficient to justify breast conservation; in 10%, tumor size did not decrease enough or increased, thus mastectomy was performed. In 11.8% of the cases, the tumor was no longer identifiable at surgical inspection, and in 3.5% no tumor was found on microscopic examination. Axillary lymph nodes were free of metastases in 39% of cases. Twelve local recurrences occurred among the 203 patients treated with breast conservation (5.9%) and five among the 23 patients treated with mastectomy (21.7%). CONCLUSIONS: Primary chemotherapy can expand the indication for breast conservation to large tumors; careful attention, however, must be paid to surgical technique. The position of the tumor should be marked with tattoo points on the skin before chemotherapy. The macroscopic extent of the tumor regression must be evaluated carefully, and multiple frozen section biopsies may be needed. The margins of the resected breast should be evaluated microscopically. All microcalcifications present before treatment must be resected. The skin incision and mammary resection must fulfill criteria of radicality as well as good cosmetic outcome. PMID- 7487209 TI - Diagnostic video-assisted thoracoscopic procedures. AB - OBJECTIVE: The authors evaluated the effectiveness and the limitations of video assisted diagnostic thoracoscopy. SUMMARY BACKGROUND DATA: The initial successes achieved with the use of video-assisted diagnostic thoracoscopic techniques has lead to an enthusiastic propagation of its use by thoracic surgeons as well as by some pulmonologists. However, detailed analyses of the diagnostic yield and potential limitations of this technique in relation to the roentgenographic and pathologic presentations of the patients are necessary to ensure its safe and effective application. METHODS: From July 1991 to December 1993, 102 diagnostic video-assisted thoracoscopic procedures were performed. All patients received other preoperative diagnostic workups without a definitive diagnosis. The initial roentgenographic presentations of these patients included 42 pulmonary nodules, 23 interstitial processes, 11 parenchymal infiltrates, 6 pleural effusions, 10 mediastinal tumors, and 10 mediastinal lymphadenopathies. If the procedure was completed without minithoracotomy or extension of any port site, then it was defined as an exclusive thoracoscopic biopsy (ETB); if the procedure was completed with the assistance of minithoracotomy (4-6 cm), then it was defined as a supplementary thoracoscopic biopsy (STB). RESULTS: Ninety-two of the pathology reports (90.2%) were interpreted as conclusive. Of these, 35 tumors were malignant and 67 benign. Ten pathology reports were inconclusive and on initial roentgenography had presented as pulmonary infiltrates (4 cases), pulmonary nodule (2), pleural effusion (2), interstitial process (1), and mediastinal lymphadenopathy (1). Seventy-six procedures (74.5%) were completed thoracoscopically and were classified as ETB. The remaining 26 procedures (25.5%) were completed with minithoracotomy and were classified as STB. The underlying diseases in the STB group were carcinoma (8 cases), Hodgkin's lymphoma (3), sarcoidosis (3), tuberculosis (2), interstitial pneumonitis (2), organizing pneumonia (2), mesothelioma (1), and miscellaneous disease (5). The reasons given for minithoracotomy were diffuse intrapleural adhesion (10 cases), technical inexperience (8), inability to localize the lesion (5), problems with anesthesia (1), poor patient tolerance (1), and unknown (1). Four patients (3.9%) experienced complications and three (2.9%) died while in the hospital. CONCLUSIONS: Diagnostic thoracoscopy provides high diagnostic yield with very low risk. However, 25.5% of the procedures require minithoracotomy to obtain adequate tissue for definitive diagnosis. This finding supports the assertion that diagnostic thoracoscopy should be performed only by experienced thoracic surgeons who can extend the procedure when indicated. PMID- 7487208 TI - Extrahepatic biliary cystadenomas and cystadenocarcinoma. Report of seven cases and review of the literature. AB - OBJECTIVE: The aim of this investigation was to describe the clinical features, diagnosis, pathologic characteristics, and optimal surgical management for patients with extrahepatic biliary cystadenomas. SUMMARY BACKGROUND DATA: Extrahepatic biliary cystadenomas are rare epithelial neoplasms. The clinical features and optimal surgical management for these lesions have not been defined clearly. The usual presenting symptom is jaundice. These lesions should be considered premalignant and necessitate resection. Sporadic case studies have reported instances of recurrence with local excision. To the authors' knowledge, this study represents the largest collected single series of extrahepatic biliary cystadenomas and reviews previously reported cases. METHODS: The authors reviewed and reported their institutional experience from 1950 to 1993 in treating seven patients with extrahepatic biliary cystadenomas as well as 19 previously reported cases in the literature. RESULTS: A strong female predominance (96.3% of patients reviewed) was associated with extrahepatic biliary cystadenomas. Obstructive jaundice was the most common presenting symptom (85%). Abdominal pain occurred in 50% of patients; other symptoms included fever and hemobilia. The most common site of occurrence was the common hepatic duct (32%). Papillary cystadenoma with foci of invasive adenocarcinoma, thus supporting the malignant potential of cystadenomas, occurred in one patient. Local excision from the wall of the bile duct was performed in 18 patients and was associated with 50% recurrence within a mean follow-up of 13 months (range, 4-24 months). No recurrence was reported after formal sleeve resection and bilioenteric reconstruction. CONCLUSIONS: Extrahepatic biliary cystadenomas can become malignant, and in this study, local surgical excision was associated with a 50% local recurrence rate. Sleeve resection with negative histologic resection margins followed by bilioenteric reconstruction, therefore, is recommended. PMID- 7487210 TI - Clinical behavior and prognostic factors of periampullary adenocarcinoma. AB - OBJECTIVE: The authors evaluated the outcome and potential prognostic factors of 60 patients with surgically resected periampullary tumors. SUMMARY BACKGROUND DATA: Periampullary carcinomas exhibit different clinical behaviors according to their site of origin. There are no prognostic factors for deciding the type of surgery to be used or for choosing patients with tumors that have a poor prognosis for adjuvant treatment. METHODS: A retrospective review was performed of 15 clinical and pathologic variables encountered among 60 patients with periampullary tumors. Tumors were divided into four groups according to their site of origin. Kaplan-Meier survival curves of the four groups were plotted and differences were evaluated with the log-rank test. Cox's proportional hazards model was used to test for separate and combined independent predictors of disease-free survival. RESULTS: Twenty-nine ampullary carcinomas, 20 ductal pancreatic carcinomas, 7 distal common bile duct carcinomas, and 4 carcinomas of the periampullary duodenum were found. Five-year disease-free survival was 43%, 0%, 0%, and 75%, respectively. According to the Cox analysis, absence of neural invasion and use of adjuvant chemotherapy were significant factors for longer survival of patients with ampullary tumors. Lymphatic invasion was related to a shorter survival in patients with pancreatic carcinoma. CONCLUSIONS: Five-year disease-free survival of patients with periampullary tumors is related to tumor type. Prognosis was better for ampullary tumors if neural invasion was absent and if adjuvant chemotherapy was used. Lymphatic invasion was associated with a shorter recurrence-free survival among patients with pancreatic carcinoma. PMID- 7487211 TI - Relation of perioperative deaths to hospital volume among patients undergoing pancreatic resection for malignancy. AB - OBJECTIVE: The authors examined the effect of hospital and surgeon volume on perioperative mortality rates after pancreatic resection for the treatment of pancreatic cancer. METHODS: Discharge abstracts from 1972 patients who had undergone pancreaticoduodenectomy or total pancreatectomy for malignancy in New York State between 1984 and 1991 were obtained from the Statewide Planning and Research Cooperative System. Logistic regression analysis was used to determine the relationship between hospital and surgeon experience to perioperative outcome. RESULTS: More than 75% of patients underwent resection at minimal-volume (fewer than 10 cases) or low-volume (10-50 cases) centers (defined as hospitals in which a minimal number of resections were performed in a given year), and these hospitals represented 98% of the institutions treating peripancreatic cancer. The two high-volume hospitals (more than 81 cases) demonstrated a significantly lower perioperative mortality rate (4.0%) compared with the minimal (21.8%) and low-volume (12.3%) hospitals (p < 0.001). The perioperative mortality rate was 15.5% for low-volume (fewer than 9 cases) surgeons (defined as surgeons who had performed a minimal number of resections in any hospital in a given year) (n = 687) compared with 4.7% for high-volume (more than 41 cases) pancreatic surgeons (n = 4) (p < 0.001). Logistic regression analysis demonstrated that perioperative death is significantly (p < 0.05) related to hospital volume, but the surgeon's experience is not significantly related to perioperative deaths when hospital volume is controlled. CONCLUSIONS: These data support a defined minimum hospital experience for elective pancreatectomy for malignancy to minimize perioperative deaths. PMID- 7487212 TI - Repeated administration of recombinant human insulin-like growth factor-I in patients after gastric surgery. Effect on metabolic and hormonal patterns. AB - OBJECTIVE: The primary objective of this investigation was to evaluate the anticatabolic effects of repeated subcutaneous administration of recombinant human insulin-like growth factor-I (rhlGF-I) in patients after gastric surgery. SUMMARY BACKGROUND DATA: The anabolic and protein-sparing effects of growth hormone are primarily mediated by IGF-I. Malnutrition and catabolic states result in increasing blood levels of growth hormone and decreasing levels of IGF-I. Experimental data showed that exogenous IGF-I could attenuate or reverse catabolism. METHODS: After giving their written informed consent, 38 male and female patients undergoing gastrectomy (age 40-75 years, body mass index 17-30 kg/m2) were treated with 80 micrograms/kg body weight rhlGF-I or placebo in a prospective, randomized, double-blind study for 5 consecutive days. Patients received a standardized total parenteral nutritional regimen with 3 g/kg body weight glucose and 0.1 g/kg body weight nitrogen. Nitrogen balance and 3 methylhistidine excretion were measured daily. Hormone profiles (IGF-I, IGFBP1, IGFBP3, cortisol, insulin, glucagon, triiodothyronine [T3], levothyroxine [T4], and thyroxine-binding globulin) were taken. PMID- 7487213 TI - Mortality and morbidity rates, postoperative course, quality of life, and prognosis after extended radical lymphadenectomy for esophageal cancer. Comparison of three-field lymphadenectomy with two-field lymphadenectomy. AB - PURPOSE: The authors evaluated the efficacy of extended radical (three-field) lymphadenectomy for esophageal cancer compared with less radical (two-field) lymphadenectomy. STUDY SUBJECTS AND ANALYTIC METHODS: The mortality and morbidity rates, postoperative courses, and survival rates were compared between 63 patients who underwent three-field lymph node dissection and 65 who underwent two field lymph node dissection at Kurume University Hospital from 1986 to 1991. Long term quality of life after surgery was compared between 37 patients who underwent three-field dissection and 35 who underwent two-field dissection from 1980 to 1991. RESULTS: Three-field dissection resulted in better survival for patients with positive lymph node metastasis from a carcinoma in the upper thoracic or midthoracic esophagus compared with two-field dissection. The mortality rates, postoperative courses and quality of life were the same for both procedures. CONCLUSIONS: Three-field dissection is preferred for upper thoracic or midthoracic esophageal cancer because of improved survival, acceptable mortality and morbidity rates, and good postoperative course and quality of life. PMID- 7487214 TI - Thoracic epidural anesthesia improves outcome after breast surgery. AB - OBJECTIVE: The authors' objective was to compare the outcomes, including the incidence of nausea and vomiting and the time until discharge to home, of patients undergoing general anesthesia and thoracic epidural anesthesia for oncologic breast procedures. SUMMARY BACKGROUND DATA: General anesthesia is the traditional anesthetic technique used in oncologic breast procedures. In March 1993, the authors initiated the use of high thoracic epidural anesthesia for patients undergoing oncologic breast surgery and reconstructive procedures. METHODS: A retrospective analysis was undertaken of 136 operations performed by one surgeon (T.J.E.) at Brigham and Women's Hospital. A chi square analysis was used to compare the outcomes of patients undergoing general anesthesia and thoracic epidural anesthesia. RESULTS: Compared with general anesthesia, thoracic epidural anesthesia was associated with a statistically significant earlier hospital discharge (p = 0.01). For quadrantectomy/axillary node dissection procedures, 20 of 39 patients (51%) having thoracic epidural anesthesia were discharged on the operative day versus 7 of 32 patients (22%) in the general anesthesia group. Furthermore, 8 of 39 patients (20%) in the thoracic epidural group experienced nausea and/or vomiting during their hospital stay versus 18 of 32 patients (56%) in the general anesthesia group (p = 0.002). CONCLUSION: Thoracic epidural anesthesia is a safe technique not associated with neurologic or respiratory complications. The use of thoracic epidural anesthesia for breast surgery could improve patients recovery and reduce the cost of these procedures. PMID- 7487215 TI - Clinical management of recurrent hepatocellular carcinoma. AB - OBJECTIVE: The aim of this study was to evaluate the long-term benefits of the aggressive treatments with resection or transarterial chemoembolization (TACE) for recurrent hepatocellular carcinoma (HCC). SUMMARY BACKGROUND DATA: Primary HCC is one of the most fatal malignancies in Taiwan. The result of resection for HCC remains unsatisfactory, primarily due to the high recurrence rate. To improve surgical results, recurrent HCC must be treated with aggressive resection or TACE. METHODS: The authors evaluated the results of repeated hepatic resection among 25 patients with recurrent HCC and of TACE among 12 patients with resectable recurrent HCC. The outcomes of an additional 64 patients with unresectable recurrent HCC were also evaluated. RESULTS: During the follow-up period from 2-112 months, 52% (13/25) of patients receiving repeat resection (group 1) were alive, whereas 42% (5/12) of patients receiving TACE (group 2) were alive. No perioperative deaths within 30 days after surgery occurred in the repeated resection group. The cumulative survival rates at 1, 2, 3, and 5 years after the first operation were 92%, 84%, 71.6%, and 65.1% in group 1 and 83.3%, 75%, 75%, and 22.5% in group 2. The survival rates at 6 months and at 1, 2, and 3 years after recurrence were 92%, 72%, 64%, and 44.8% in group 1 and 83.3%, 75%, 66.7%, and 48% in group 2. The survival of patients with unresectable recurrent HCC was much worse: 1-, 2-, 3-, and 5-year survival after surgery was 57.8%, 29.8%, 15.5%, and 0%; and 6-month and 1-, 2-, and 3-year survival after recurrence was 46.5%, 29.2%, 12.5% and 7.8%. CONCLUSIONS: More aggressive treatment with repeated hepatic resection can prolong survival time after recurrence of HCC in selected patients. However, TACE can also achieve good results although it is not thought of as curative. PMID- 7487216 TI - Laparoscopic aortofemoral bypass. Initial experience in an animal model. AB - OBJECTIVE: The study objective was to evaluate the feasibility of laparoscopic aortofemoral bypass in a porcine model. SUMMARY BACKGROUND DATA: Laparoscopic techniques have been applied to numerous general and thoracic surgical procedures. Their application to vascular surgery has been virtually nonexistent. Open surgery for aortoiliac occlusive disease is accompanied by significant morbidity rates, and minimally invasive procedures have the disadvantage of reduced patency rates. Laparoscopic aortofemoral replacement has the theoretical advantage of long-term patency with reduced postoperative complications. METHODS: Between January and September 1993, laparoscopic surgery was performed on 16 pigs: 6 underwent transperitoneal laparoscopic aortic dissection and vessel control alone; 7 underwent complete transperitoneal laparoscopic aortofemoral bypass; and 3 underwent a retroperitoneal approach. The aortic anastomosis was performed using a combination of sutures and titanium clips in an end-to-side fashion in five pigs, and a custom-made nonsutured graft was secured with use of an end-to-end method in five pigs. Femoral anastomoses were performed with the standard open technique. RESULTS: Technical success was achieved in all 10 animals and with no major complications. Mean blood loss was 20 ml (range, 5-50 ml), and mean operative time was 2.45 hours (range, 2-4 hrs). On aortic-clamp release, 2 of the end-to-side anastomoses required additional sutures to stop bleeding between oversized staples, and 2 of the end-to-end anastomoses required additional ties to reinforce loose ties. All 10 grafts and anastomoses were patent and free of leaks after completion of the procedure. CONCLUSIONS: Laparoscopic aortofemoral bypass is technically feasible in a porcine model. Further experimental work with new instrumentation and technical refinement will make laparoscopic surgery feasible for the treatment of vascular disease in humans. PMID- 7487217 TI - Selected patients with very small invasive breast cancers can be treated with breast preservation. PMID- 7487218 TI - The operative mortality rate in the R3 total gastrectomy group. PMID- 7487219 TI - Earlier methods of antibiotics administered prophylactically perioperatively. PMID- 7487220 TI - Documenting 51 cases of laparoscopic colectomies from two institutions and six different surgeons. PMID- 7487222 TI - Trapping the flies that cause myiasis: their responses to host-stimuli. AB - A critical phase in the life cycle of the fly species that cause myiasis is host location by the adult females, whether by direct location of and contact with the host, or by indirect location through the selection of sites frequented by hosts. The stimuli to which these flies respond in host location, principally visual and olfactory, are discussed here in the context of their use to enhance the performance of trapping systems used in research, monitoring and control. The three major families of flies encountered in myiasis can be divided into two groups for comparison of host-location strategies: (1) Oestridae; and (2) Calliphoridae and Sacrophagidae. The former are all obligate parasites, the latter include obligate and primary facultative parasites. In the Oestridae, the majority of Cuterebrinae deposit their eggs at sites likely to be visited by the host, whereas the majority of Oestrinae, Hypodermatinae and Gasterophilinae deposit their eggs or larvae directly onto the host. It is presumed that most host location is, like that of haematophagous insects, by response to olfactory and visual cues released by healthy hosts, but the subject is very under researched. In the Calliphoridae and Sacrophagidae, most species that cause myiasis deposit their eggs or larvae directly onto the host at some predisposing site, such as those caused by wounding, necrosis and/or bacterial contamination. Healthy hosts are much less attractive, even for obligate species such as Wohlfahrtia magnifica, which can invade undamaged tissues. The last decade has seen a renewed interest in the potential for using traps in control of myiasis, leading to significant advances in trap and bait efficiency. However, there is great potential for further research and development, particularly on the general responses of Oestridae to hosts and in the precise identification of the olfactory cues for Calliphoridae and Sarcophagidae. PMID- 7487221 TI - 15 patients with mucocele of the gallbladder in the acute cholecystitis group. PMID- 7487223 TI - The epidemiology of malaria in the Wosera area, East Sepik Province, Papua New Guinea, in preparation for vaccine trials. I. Malariometric indices and immunity. AB - The epidemiological features of malaria were studied through seven community based surveys in a population of 4000 in the Wosera area, East Sepik Province, Papua New Guinea. Prevalence of parasitaemia (all species, all ages) was 60%. Plasmodium falciparum was the predominant species in all surveys (55%), followed by P. vivax (25%) and P. malariae (20%). The highest prevalence for asexual forms of P. falciparum occurred in the 5-9-year age group, whereas P. falciparum gametocytaemia and P. vivax parasitaemia were observed most frequently in the 1-4 year age group and P. malariae in the 10-15-year age group. Mean densities of all species decreased with age except for that of P. malariae, which was lower in children aged < 1 year than in those aged 1-4 years. The prevalence of enlarged spleen was 57% in children and 10% in adults and closely matched the corresponding age-related parasite rate. Seroprevalence of antibody to the major merozoite surface antigen 2 rapidly increased with age, with > 90% of individuals older than 5 years being positive. Malariological indices showed irregular changes over time but there was no clear-cut seasonal pattern. The geographical distribution of these indices and immune responses was not uniform within the study area. Bednet use and drug consumption were negatively correlated with malariometric indices. Identification of significant temporal and local variations in malaria endemicity is important for the design and evaluation of intervention studies, including field trials of an antimalarial vaccine. PMID- 7487224 TI - The epidemiology of malaria in the Wosera area, East Sepik Province, Papua New Guinea, in preparation for vaccine trials. II. Mortality and morbidity. AB - Malaria mortality and morbidity were studied in a rural population of 4000 in the Wosera area, East Sepik Province, Papua New Guinea. Malaria accounted for 4.9% of the 162 deaths investigated by verbal autopsy and for 12.2% of the 49 deaths assessed through medical records. Malaria was the first cause of death in children aged 0.5-4 years. Of the 7795 subjects interviewed and bled during six cross-sectional community-based surveys, children of 1-4 years had the highest malaria-related morbidity. In this age group, point prevalences of fever, fever associated with parasitaemia, and fever plus Plasmodium falciparum (Pf) parasitaemia > or = 10,000 parasites/microliters blood were 5%, 4.1% and 1.5%, respectively. The corresponding figures for adults were 2%, 0.9% and 0.1%, respectively. The calculation of attributable fraction (AF) using a multiple logistic regression model showed that malaria accounted for 0.44 of all fevers in children of 1-4 years and 0.08 of the fevers in adults. Prevalence data derived from the AF estimate were compared with those calculated using different accepted density thresholds. The prevalences which best approximated the results from the logistic regression model were obtained using parasitaemia cut-offs of > or = 1000 Pf parasites/microliter in children aged 1-4 years and adults older than 19 years and of > or = 10,000 parasites/microliter in those aged 5-19 years. Prevalence of fever associated with parasitaemia was highly seasonal, with a peak at the beginning of the wet season. The geographical distribution of malaria morbidity was not uniform. The measurement of malaria-related morbidity, the identification of significant seasonal and local variation as well as the assessment of different methods of defining a clinical episode of Pf malaria are crucial for the design and evaluation of intervention studies, including field trials of antimalarial vaccines. PMID- 7487225 TI - Elevated 2,3-diphosphoglycerate concentrations and alteration of structural integrity in the erythrocytes of Indian cases of visceral leishmaniasis. AB - The visceral leishmaniasis (VL) known as kala-azar in India is characterized by severe anaemia. The anaemia seems to be the result, at least in part, of the relatively short life-time of the erythrocytes, which have weakened cell membranes, possibly because of elevated concentrations of 2,3-diphosphoglycerate (2,3-DPG). There is a negative correlation (r = 0.91; P < 0.01) between erythrocytic 2,3-DPG concentrations and the blood concentration of haemoglobin, and the erythrocytes from infected patients display higher osmotic fragility than those from uninfected controls. Spectrofluorometry, using 1,6-diphenyl 1,3,5 hexatriene as a probe, indicated that fluorescence depolarization and microviscosity are also higher in the erythrocytic membranes from VL cases than in those from the controls. The cholesterol/phospholipid ratio is also relatively high in the membranes from the VL cases and there is degradation of the skeletal components and the major integral protein (band 3). The enhanced concentration of 2,3-DPG may be related to the altered structural integrity of the erythrocytes and this may lead to anisocytosis and the reduction in the erythrocytic half life. PMID- 7487226 TI - Taenia solium cysticercosis and human taeniasis in the Nsukka area of Enugu State, Nigeria. AB - The prevalence of Taenia solium cysticercosis in slaughter pigs and of taeniid ova in hospital patients were determined in the Nsukka area of Enugu State, Nigeria, in March 1986-September 1988 and May 1986-May 1988, respectively. Cysticercus cellulosae were detected in the pigs by ante-mortem examination of the pigs' tongues and detailed post-mortem examination of the dressed pig carcasses using standard meat-inspection procedures. Human infection was assessed by examining iodine-stained stool samples collected from patients from one selected hospital in the study area. Over 20% (483) of the 2358 trade pigs examined were found infected with C. cellulosae. Most of the cases were generalized, all the musculature being heavily infested with live cysticerci. The age and sex of the pig and the season of the year in which it was examined had no significant effect on the occurrence of cysticerci in the animals (P > 0.01) but there was a highly significant year-to-year decrease (P < 0.001) in the prevalence of cysticerci. The overall prevalence of taeniid ova in the 1525 human stool samples analysed was 8.6%, most (78.6%) of the cases occurring in adults aged > 30 years. The epidemiological factors which might have influenced these results are identified and discussed, and suggestions are made for the control of this important zoonosis. PMID- 7487227 TI - A retrospective study of human cystic echinococcosis in Jordan. AB - Some 676 human cases of cystic echinococcosis (CE) were recorded from 18 major hospitals in Jordan from January 1985 to October 1993. The number and origins of the cases indicate that CE is an endemic and widespread disease in Jordan. Operations for the extraction of hydatid cysts represented 0.1% of all surgical operations carried out in the surveyed hospitals during the study period (excluding Caesarean sections). The mean annual surgical incidence of CE was estimated to be 0.5-8.2 per 100,000 population, depending on the governorate studied, with an overall mean of 2.9 per 100,000. Incidence was higher in the more rural eastern and south-western governorates than elsewhere, and more cases fell in the 21-30-year age group than in any other. Many more of the cases were female than male (443 v. 233; P < 0.001). The liver was the most common organ infected (54.5% of cases), followed by the lungs (12.1%), and multiple hydatidosis usually involved the liver. PMID- 7487229 TI - Secondary cutaneous manifestations of acute schistosomiasis mansoni. AB - The secondary cutaneous manifestations of the initial phase of schistosomiasis mansoni were studied in 34 patients who had been infected simultaneously in the same location. Sixteen of the patients developed angioedema and/or urticaria, generally of short duration and of mild intensity and usually about 30 days post infection. There was no apparent association between the occurrence of these skin manifestations and the patients' worm burdens, blood eosinophilias or areas of immediate reaction to an intradermal inoculation with worm antigen. Other factors, particularly host immunological reactivity, are thought to be important elements in the genesis of the manifestations. PMID- 7487228 TI - Immune response of mice infected with Schistosoma mansoni is modulated by antifibrotic treatment. AB - Mice infected with Schistosoma mansoni treated with beta-aminopropionitrile (BAPN), an antifibrotic agent, and the antischistosomal drug praziquantel (PZQ) were resistant to challenge for up to 5 weeks post-treatment. The combined treatment resulted in profound changes to the liver granuloma cell matrix and the composition and function of the cellular infiltrate. Although granulocytes always predominated in the infiltrate, the proportion of the cells which were macrophages was higher in mice treated with BAPN alone (39.2%) than in infected mice which were untreated (15.2%) or treated with PZQ alone (12.4%), and much higher than in mice given the combined treatment (1.8%). Two products associated with macrophage activation and cytotoxicity [tumour necrosis factor-alpha (TNF alpha) and nitric oxide] were only detected in mice harbouring a patent infection and there was a strong positive correlation between the concentrations of each. The relatively low TNF-alpha concentrations in BAPN-treated mice seemed to be associated with the relatively small granulomas observed in these mice. BAPN treatment also led to changes in the proliferative response of the treated mice's macrophages to mitogen and soluble schistosome-egg antigen and in spleen cellularity; these changes are probably associated with the resistance to challenge infection observed in mice given BAPN with PZQ. It is clear that BAPN treatment changes the dynamics of the delayed-type hypersensitivity response within the granuloma and that this impacts on other immunological sites. How this relates to the maintenance of post-treatment resistance to a challenge infection has still to be elucidated. PMID- 7487230 TI - Profile of hepatitis C virus and the possible modes of transmission of the virus in the Gizan area of Saudi Arabia: a community-based study. AB - The seroprevalence of antibody to hepatitis C virus (anti-HCV) and the possible modes of transmission of HCV were investigated in Gizan, southern Saudi Arabia. The sample size chosen to give an adequate estimate of the seroprevalence, about 1500, was based on the assumption that 5% of the population in Gizan were anti HCV-positive. Sera from 1482 subjects (705 males, 777 females; aged > or = 10 years) were initially screened for anti-HCV using a commercial, ubiquitin-based enzyme immunoassay. Repeatedly reactive sera were confirmed positive using second generation immunoassays. Serum samples were also tested by ELISA for hepatitis B surface antigen (HbsAg) and antibodies to this antigen and to the hepatitis B core antigen. Of the subjects tested, 27 (1.8%) were anti-HCV-positive. Exposure to HCV was generally similar in both sexes, age-prevalence curves for anti-HCV peaking in males aged > 49 years (6.2%) and in females aged 40-49 years (5.0%). In the youngest subjects, those aged 10-19 years, the HbsAg carrier rate was significantly higher in males (10.4%) than in females (3.6%). Exposure to the hepatitis B virus was similar in both sexes (31.0% in males v. 28.6% in females). Some 7.4% and 14.8% of the 27 anti-HCV-positive cases had histories of schistosomiasis and blood transfusion, respectively. The corresponding values for the 1455 anti-HCV-negative cases investigated, 1.1% for schistosomiasis and 3.5% for blood transfusion, were much lower. The spouses and other family members of eight anti-HCV-positive index cases were investigated but none was anti-HCV positive.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7487231 TI - Validation of schistosomiasis morbidity symptoms in schoolchildren of Siavonga, Lake Kariba, Zambia. PMID- 7487232 TI - Evaluation of a recombinant antigen-based antibody assay for diagnosis of bancroftian filariasis in Egypt. PMID- 7487233 TI - Arteether neurotoxicity in the absence of deficits in behavioural performance in rats. PMID- 7487234 TI - Calcium antagonists and mortality. Another case of the need for clinical judgment. PMID- 7487235 TI - Cholesterol screening. Once is not enough. PMID- 7487236 TI - Calcium antagonists should continue to be used for first-line treatment of hypertension. PMID- 7487237 TI - Should dihydropyridines be used as first-line drugs in the treatment of hypertension? The con side. PMID- 7487238 TI - Guidelines for the diagnosis and treatment of gastroesophageal reflux disease. Practice Parameters Committee of the American College of Gastroenterology. AB - Gastroesophageal reflux disease (GERD) is a common disorder that affects all segments of the population. These guidelines have been prepared to suggest the preferable, but not only, approaches to the management of a patient with GERD. The physician must be free to choose the course best suited to the individual patient. The world literature was reviewed and the guidelines were developed under the auspices of the American College of Gastroenterology, and approved by other gastroenterology societies. Diagnostic recommendations include the following: (1) general approach to GERD, including empiric therapy, (2) appropriate mucosal evaluation (ie, endoscopy and radiology), (3) use of pH and provocative testing, and (4) indications for manometric evaluation. Therapeutic recommendations include the following: (1) general approach to therapy including lifestyle changes, (2) use of acid suppression, (3) use of promotility drugs, (4) maintenance treatment of GERD, and (5) indications for antireflux surgery. PMID- 7487239 TI - Evaluating cholesterol screening. The importance of controlling for regression to the mean. AB - OBJECTIVES: To determine the overall impact of a voluntary cholesterol screening program and to assess the importance of controlling for regression to the mean when evaluating the impact on higher-risk subgroups. DESIGN: Longitudinal study comparing baseline survey and cholesterol results with those obtained 17 months later, with adjustment for regression to the mean. SETTING: Participants were seen at three sites: an academic teaching hospital, an intermediate-care hospital, and a suburban community center. PARTICIPANTS: One thousand fifty-three participants were enrolled in a voluntary cholesterol screening program. MEASUREMENTS/MAIN RESULTS: Of 1053 study enrollees, 785 (75%) completed a follow up questionnaire and 656 (62%) returned for follow-up cholesterol tests. Among all participants at high risk, measured cholesterol levels were 0.66 mmol/L (25.5 mg/dL) lower at follow-up, but 58% of this apparent change (0.38 mmol/L [14.7 mg/dL]) was attributable to the statistical phenomenon of regression to the mean. Among all participants at increased (both high and moderate) risk, after controlling for regression to the mean, reductions in cholesterol levels were significant for those younger than 60 years (-0.28 mmol/L [-10.8 mg/dL]; 95% confidence interval, -0.43 to -0.13; n = 390) but not for those 60 years or older (+0.007 mmol/L [+0.3 mg/dL]; 95% confidence interval, -0.16 to +0.17; n = 266). CONCLUSIONS: For subjects found at high risk in a cholesterol screening program, more than half of the apparent beneficial change in cholesterol level is attributable to regression to the mean. For participants older than 60 years, no clear benefit of cholesterol screening was documented. PMID- 7487240 TI - Long-term patient self-management of oral anticoagulation. AB - BACKGROUND: The management of oral anticoagulation is fraught with difficulties. This study assessed a new model of anticoagulation management regarding the ability, safety, and efficacy of patients to self-monitor and self-adjust the dose of their oral anticoagulants guided by a capillary whole-blood prothrombin time (PT) monitor. METHODS: This investigation is a retrospective cohort study of 20 patients compared with 20 matched control patients receiving oral anticoagulation at a tertiary medical institution. RESULTS: Study patients monitored their PTs 2153 times during a mean interval of 44.7 months compared with 1608 PTs in matched control patients during a mean interval of 42.5 months. Study patients made an average of 11.5 dosage changes per patient, contrasted with 22.7 changes per control patient (P < .001). The PTs in study patients were within the recommended therapeutic range in 88.6% (95% confidence interval, 87.2 to 89.9) of the determinations compared with 68.0% (95% confidence interval, 65.7 to 70.3; P < .001) of the determinations made by the matched control patients. In response to the 2153 PTs, study patients made 67 (3.1%) dosage decisions that were considered incorrect based on physician guidelines. None of these changes led to adverse outcomes. There was no significant difference in complication rates between the two groups. CONCLUSIONS: Results from what is the first long term study of patient self-monitoring of PTs and self-adjustment of the warfarin sodium dosage for oral anticoagulation suggest that patients can successfully measure their own PTs, adjust their own warfarin dosage, and achieve a degree of therapeutic effectiveness at least as good, if not better than patients managed in an anti-coagulation clinic. Larger, prospective, randomized trials are needed to confirm the efficacy and safety of this new approach to therapy and to assess its cost-effectiveness. PMID- 7487241 TI - Prevalence of residual left atrial thrombi among patients with acute thromboembolism and newly recognized atrial fibrillation. AB - BACKGROUND: Thromboembolism related to atrial fibrillation (AF) is a major cause of morbidity and mortality. Patients with acute thromboembolism and AF are at high risk for early recurrent events. OBJECTIVE: To determine the prevalence of left atrial thrombi in patients who had acute thromboembolism and newly diagnosed AF. PATIENTS AND METHODS: Adult inpatients with AF were screened to identify those with acute (< 36 hours) systemic thromboembolism and newly recognized AF. Of 41 qualifying patients, 31 (76%) agreed to undergo transesophageal echocardiographic study, including 24 with acute neurologic events and seven with peripheral thromboembolism. A control population consisted of 88 adults with newly recognized AF without clinical thromboembolism. RESULTS: Transesophageal echocardiography identified left atrial thrombi in 13 (43%) of the 30 study patients who underwent transesophageal echocardiography compared with nine (10%) of 87 controls (P < .001). Spontaneous echo contrast was identified in 27 (87%) of the study population vs 42 (48%) of controls (P < .001). The prevalence of this marker of blood stasis did not differ between patients with left atrial thrombi without thromboembolism (P = .69). Duration of AF, prevalence of abnormal left ventricular function, left atrial size, and mitral regurgitation were similar in both groups. CONCLUSIONS: Left atrial thrombi were identified in more than 40% of patients with acute thromboembolism and newly recognized AF. These data suggest that a major source of recurrent thromboembolism in this group may be residual thrombus migration. Among patients with AF and atrial thrombi, clinical thromboembolism seems to occur randomly, or is related to an unidentified process. PMID- 7487242 TI - Promoting completion of health care proxies following hospitalization. A randomized controlled trial in a community hospital. AB - BACKGROUND: The wider use of written advance directives is popular but problematic. We have shown previously that acute hospitalization in the era of the Patient Self-Determination Act can facilitate directive discussions and documentation. We investigated whether a simple educational intervention following hospitalization would increase patients' execution of durable health care proxies. METHODS: We studied a consecutive series of patients (n = 162) recently discharged from the acute care medical service of a community hospital where they had been interviewed about advance directives. The intervention group was randomized to receive an educational brochure and encouragement to execute durable health care proxies. The primary outcome was the proportion of patients in each group with completed durable health care proxies on file in their primary physicians' offices. RESULTS: Overall, only 20 (12.3%) of 162 patients had documented proxies, 17 of whom (85%) were 65 years of age or older, with no difference between the intervention and control groups (11 [13.3%] of 83 vs nine [11.4%] of 79, respectively). Subgroup analysis of elderly patients also revealed no intervention effect. Univariate analysis revealed three significant predictors of patients' proxy completion: patient age, whether patients had discussed directives in hospital with their physicians, and whether patients' physicians completed proxies for themselves. Multiple logistic regression analysis showed that these three variables interact to predict the probability of patients' executing proxies. CONCLUSIONS: Simple educational interventions, like those mandated by the Patient Self-Determination Act, are unlikely to increase patients' completion of durable healthcare proxies. Multidimensional interventions that target both elderly patients and their personal physicians should be tested in the future. Discussion in hospital about advance directives can be a useful component of such efforts. PMID- 7487243 TI - Putting prevention into practice. Impact of a multifaceted physician education program on preventive services in the inner city. AB - BACKGROUND: Physicians' prevention practices often differ from guidelines published by national authorities. Effective preventive services are most needed in inner city settings that suffer disproportionately from preventable diseases. This study examined the impact of a multifaceted physician prevention education program on the provision of preventive services in an inner city municipal hospital. METHODS: The study used a controlled intervention comparative design at two inner city municipal hospitals--Harlem Hospital Center, New York, NY (intervention site) and Kings County Hospital, Brooklyn, NY (comparison site)- serving predominantly African-American patient populations. The intervention site received prototype materials for physicians, patients, and the office setting from the US Public Health Service's Put Prevention Into Practice campaign and a series of prevention lectures from November 1991 through April 1992. Change in physician prevention practices and knowledge was assessed by self-administered questionnaires and change in patients' reports of preventive services received was assessed by structured interviews. RESULTS: Physicians at Harlem Hospital Center reported a greater postintervention increase in prevention practices and demonstrated a greater increase in prevention knowledge in comparison with physicians at Kings County Hospital. Patients at Harlem Hospital Center reported receiving increased preventive services from physicians after the intervention, while patients at Kings County Hospital did not report any significant change in preventive services received. CONCLUSIONS: A multifaceted physician education program using prototype materials from the Put Prevention Into Practice campaign with prevention lectures significantly increased the prevention knowledge and practices reported by physicians and the preventive services reported received by patients at an inner city municipal hospital. PMID- 7487244 TI - Clinical evaluation of different doses of intravenous enalaprilat in patients with hypertensive crises. AB - BACKGROUND: The appropriate dose of intravenous enalaprilat to be used in the treatment of hypertensive crisis is controversial. There has been no comparative study of the efficacy and safety of different dosages of enalaprilat in hypertensive patients. METHODS: Sixty-five consecutive patients with hypertensive urgencies (systolic blood pressure > 210 mm Hg and/or diastolic blood pressure > 110 mm Hg) or emergencies (diastolic blood pressure > 100 mm Hg and evidence of end-organ damage, ie, angina pectoris, hypertensive encephalopathy, or congestive heart failure) admitted to an emergency department from January 1, 1994, to September 30, 1994, were identified. The patients were randomized to receive different doses of enalaprilat (0.625, 1.25, 2.5, and 5 mg). Response to treatment was defined as a stable reduction of systolic blood pressure to below 180 mm Hg and diastolic blood pressure to below 95 mm Hg within 45 minutes after the start of treatment and relief of symptoms in patients with hypertensive emergencies. RESULTS: In 41 (63%) of 65 patients, the treatment goal was reached. Twenty-four patients (37%) failed to achieve the goal of treatment within 45 minutes after administration of enalaprilat. The response rates in the 0.625-mg, 1.25-mg, 2.5-mg, and 5-mg groups were 67%, 65%, 59%, and 62%, respectively. The proportion of patients initially randomized who responded to treatment was not different between any of the four groups of enalaprilat doses. There were no significant differences according to enalaprilat dose with respect to changes in systolic, diastolic, and mean arterial blood pressure. No severe side effects were observed. CONCLUSION: Enalaprilat is a safe antihypertensive drug with moderate efficacy in the treatment of hypertensive crisis. As doses above 0.625 mg alter neither response rates nor the magnitude of blood pressure reduction, we recommend 0.625 mg as the initial dose in the treatment of hypertensive crisis. PMID- 7487245 TI - Effect of pulmonary function testing on the management of acute asthma. AB - BACKGROUND: Routine pulmonary function testing has been recommended as part of the emergency department management of acute asthma despite the lack of evidence demonstrating the necessity for these measurements. The purpose of this study was to assess the physician's ability to estimate pulmonary function in patients with acute asthma and to determine the effect that pulmonary function testing has on patient management. METHODS: Emergency medicine physicians estimated pulmonary function in adult asthmatic patients prior to spirometry. The estimations and spirometry were repeated after 1 hour and again at discharge. The physicians noted their treatment plan before and after receiving the results of pulmonary function testing. RESULTS: Ninety-eight patients were enrolled in the study. There was only moderate correlation between pretreatment estimates of pulmonary function and the actual value (r = .41). On average, physicians underestimated the degree of pretreatment airway obstruction (as percentage of predicted normal value) by 8.1 +/- 16.0 percentage points. Knowledge of the pulmonary function test results changed management in 20.4% of patients. The most common change was an alteration in the decision to continue treatment after 3 hours. CONCLUSIONS: Physicians tend to underestimate the degree of airway obstruction in acute asthma, particularly on initial assessment. Since the availability of pulmonary function tests changes management in a significant number of patients, routine pulmonary function testing is recommended as part of the assessment and monitoring of acute asthma. PMID- 7487247 TI - Anticholinergic poisoning associated with herbal tea. AB - An outbreak of cholinergic poisoning occurred in New York City during a 3-day period. Seven individuals from three families of South American origin were affected. Signs and symptoms of illness included dry skin, hyperthermia, tachycardia, dilated pupils, agitation, and hallucinations. Onset of illness in all cases was temporally associated with consumption of a tea that was labeled "Paraguay Tea" and was purchased from a grocery store specializing in South American foods. Paraguay tea, made from the leaves of the holly, Ilex paraguariensis, contains caffeine and theophylline and is a popular beverage in South America. Samples of the tea analyzed with gas chromatography contained belladonna alkaloids but neither caffeine nor theophylline. An investigation by the New York City Department of Health personnel determined that the tea was from a single lot, imported by one distributor, and sold at one grocery store. Unsold inventories of the tea were quarantined, and no further cases of anticholinergic poisoning were reported. PMID- 7487246 TI - Clinical correlates of secondary meningitis in HIV-infected adults. AB - BACKGROUND: Neurologic complaints are common in adults infected with the human immunodeficiency virus, but little is known about which clinical features are associated with secondary causes of meningitis. METHODS: A retrospective cross sectional study of adults infected with the human immunodeficiency virus who received a diagnostic lumbar puncture (LP) in the infectious disease clinic, emergency department, and inpatient wards of the Deaconess Hospital, Boston, Mass, from 1989 through 1992 to determine which clinical features available at the time of LP are correlated with definite or probable secondary meningitis. RESULTS: Of the 491 LPs, 90% were performed in whites, 93% in men, and 11% in injection drug users. Cerebrospinal fluid test results revealed secondary meningitis in 39 (7.9%) of 491 LPs performed on 322 individuals. Cryptococcal meningitis was the predominant type (27 cases); no bacterial or tuberculous meningitis was found. In multivariate analyses, a history of non-Hodgkin's lymphoma (adjusted odds ratio [OR], 4.3; 95% confidence interval [CI], 1.5 to 12.5), a history of herpes simplex virus infection (OR, 2.5; 95% CI, 1.2 to 5.0), nausea and/or vomiting (OR, 2.0; 95% CI, 1.03 to 4.0), headache in a person with the acquired immunodeficiency syndrome (OR, 2.1; 95% CI, 1.03 to 4.4), and cranial nerve abnormalities (OR, 5.1; 95% CI, 1.8 to 14.1) were positive correlates of opportunistic meningitis; current fluconazole use (OR, 0.3; 95% CI, 0.1 to 0.8) conferred a lower risk. CONCLUSION: In similar clinical settings, physicians and their human immunodeficiency virus-infected patients should consider these features when assessing the risk of secondary meningitis and the necessity for immediate LP. PMID- 7487248 TI - Death by destruction of will. Lest we forget. AB - This report describes an unexplained death following the collapse of will caused by resisted mechanical restraint in a 93-year-old woman with dementia. Clinical and experimental evidence is cited supporting resisted restraint as a significant noxious stimulus that can be linked to sudden death and immune dysfunction. This woman's story serves to remind us of the critical link between mind and body, and of the mortal consequences possible when we forget. PMID- 7487249 TI - Nonulcer dyspepsia and Helicobacter pylori: causal or coincidental? PMID- 7487250 TI - On advancing advance directives. Why should we believe the promise? PMID- 7487251 TI - In the eye of the beholder. PMID- 7487252 TI - Anticonvulsant hypersensitivity syndrome. AB - Anticonvulsant hypersensitivity syndrome is a potentially fatal drug reaction with cutaneous and systemic reactions (incidence, one in 1000 to one in 10,000 exposures) to the arene oxide-producing anticonvulsants--phenytoin, carbamazepine, and phenobarbital sodium. In most cases, the hallmark features of fever, rash, and lymphadenopathy are accompanied by multiorgan-system abnormalities. Fatal outcomes are most often associated with liver failure. Recognition of the syndrome, which may have variable presentations, is the key to prompt discontinuation of the drug, close monitoring, and management. The reaction may be genetically determined, and siblings of patients with anticonvulsive hypersensitivity syndrome may be at increased risk of developing this syndrome. The timely recognition of anticonvulsant hypersensitivity syndrome is important, because accurate diagnosis avoids potentially fatal reexposure and affects subsequent anticonvulsant treatment options. PMID- 7487253 TI - A new typhoid vaccine composed of the Vi capsular polysaccharide. AB - Typhoid is still prevalent in many parts of the world. We reviewed all published and unpublished studies of a newly licensed vaccine composed of the Vi capsular polysaccharide of Salmonella typhi, the causative agent of the disease, which had been licensed previously outside the United States. These included observational studies and double-blind randomized studies done in the United States, Europe, and the developing world in which children and adults unexposed to typhoid or those living in endemic areas were enrolled. A single dose of 25 micrograms of the purified polysaccharide was given by intramuscular injection. The vaccine was well tolerated, inducing only minor reactions in fewer than 10% of subjects. An antibody response occurred in about 90% of subjects and lasted about 3 years. Seroconversion was shown in children as young as 2 years. Protective efficacy was evaluated in two studies conducted in areas in which typhoid is endemic; the efficacy was 55% and 75%, respectively, in adults and in children older than 5 years. The Vi vaccine compares favorably with other typhoid vaccines in regard to safety, patient compliance, immunogenicity, and efficacy. Vi polysaccharide is a well-standardized antigen that is effective in a single parenteral dose, is safer than whole-cell vaccine, and may be used in children 2 years of age or older. PMID- 7487254 TI - Leisure-time physical activity and the risk of nonfatal myocardial infarction in postmenopausal women. AB - BACKGROUND: Few studies have examined the association of physical activity with coronary heart disease among women. OBJECTIVE: To examine whether participation in physical activity during leisure time decreases the risk of myocardial infarction in postmenopausal women. METHOD: A population-based, case-control study among enrollees of the Group Health Cooperative of Puget Sound, a health maintenance organization based in Seattle, Wash. Cases were postmenopausal women who sustained an incident nonfatal myocardial infarction during the period 1986 through 1991. Controls were a random sample of Group Health Cooperative enrollees who were frequency matched to the cases by age and calendar year. Participation in physical activity during leisure time was assessed from a telephone interview. A total of 268 cases and 925 controls were interviewed. RESULTS: After adjustment for potential confounding factors, the odds ratios for nonfatal myocardial infarction for women in the second, third, and fourth quartile of total energy expenditure, relative to women in the first quartile, were 0.52 (95% confidence interval, 0.34 to 0.80), 0.40 (95% confidence interval, 0.26 to 0.63), and 0.40 (95% confidence interval, 0.25 to 0.63), respectively. Similar odds ratios were associated with the energy expended in nonstrenuous leisure-time physical activity, and with walking for exercise. CONCLUSION: This case-control study suggests that the risk of myocardial infarction among postmenopausal women is decreased by 50% with modest leisure-time energy expenditures, equivalent to 30 to 45 minutes of walking for exercise three times a week. PMID- 7487255 TI - The influence of clinical risk factors on the use of angiography and revascularization after acute myocardial infarction. Myocardial Infarction Triage and Intervention Project Investigators. AB - BACKGROUND: Coronary revascularization provides the greatest survival advantage in those patients with the greatest mortality risk. This study examines the relationship between variables that predict mortality and the use of angiography and revascularization after acute myocardial infarction. METHODS: Study of 4823 survivors of acute myocardial infarction, who underwent angiography between 6 hours and 5 days of admission, to determine the relationship between factors that predict mortality and the use of angiography (n = 2274), angioplasty (n = 692), and bypass surgery (n = 469). RESULTS: Except for recurrent angina, clinical factors that predict higher mortality were associated with a lower use of angiography (the multivariable adjusted odds ratio was 0.47 for older age, 0.85 for a history of infarction, 0.50 for patients not receiving thrombolytic medications, 0.64 for new heart failure, and 2.75 for recurrent angina [P < .001 for all factors]). A similar relationship was observed among patients selected for angioplasty (the odds ratio was 0.51 for an ejection fraction of < 40%, 0.72 for those patients not receiving thrombolytic medications, 0.74 for a history of infarction, and 1.94 for recurrent angina [P < .001 for all factors]). In contrast, patients with unfavorable prognostic profiles were much more likely to undergo coronary bypass surgery (the odds ratio was 1.46 for recurrent angina, 1.28 for older age groups, 2.23 for new heart failure, 1.28 for patients not receiving thrombolytic medications, and 1.46 for a history of infarction [P < .001 for all factors]). CONCLUSIONS: These data suggest that aside from symptoms of recurrent angina, the use of angiography and angioplasty is not driven by mortality risk stratification. In contrast, bypass surgery is preferentially performed in patients at increased risk for mortality. PMID- 7487256 TI - Promoting inpatient directives about life-sustaining treatments in a community hospital. Results of a 3-year time-series intervention trial. AB - BACKGROUND: Hospitalization presents an opportunity for physicians to discuss advance directives with patients and to encourage completion of health care proxies. OBJECTIVE: To prospectively promote discussion and documentation of treatment-specific directives about life-sustaining interventions (cardiopulmonary resuscitation, admission to critical care units, mechanical ventilation, electrical cardioversion, and vasopressor therapy) among unselected medical inpatients in a community teaching hospital. METHODS: We conducted a time series intervention trial from January 1, 1991, through June 30 1993, divided into three phases. During the education phase, we provided reminders, education, and feedback to attending physicians; during the intervention phase, we promoted a new documentation form for directives to be used by attending physicians; during the control phase, no interventions occurred. We studied consecutive patients (N = 1780) admitted to the hospital acute medical service in each of the following 10 periods: three in the education phase (n = 598), three in the intervention phase (n = 826), and four in the control phase (n = 356). The primary outcome measures were the frequency and content of directives documented by attending physicians in their patients' hospital charts. Secondary outcome measures included physicians' and patients' attitudes about directives, surveyed repeatedly. RESULTS: The proportion of inpatients with directives increased significantly during the intervention phase (62.5% vs 23.6% during the education phase and 25.3% during the control period, P < .001, Pearson chi 2 test). During the final intervention phase, 227 (83.2%) of 273 inpatients had directives documented in the hospital chart. Increases in clinically important ("impact") directives usually involved intensive care, not do-not-resuscitate status. Overall, 366 (86.7%) of 422 physician-attested directives agreed with the treatment preferences of interviewed patients (kappa ranges, 0.53 to 0.79). Physicians' attitudes about and interest in directives improved. CONCLUSIONS: Institutional interventions can facilitate attending physicians' documentation of treatment-specific directives about life-sustaining care for most medical inpatients. More research is needed to confirm the effect of these efforts on quality and cost of hospital care, patients' autonomy, and their eventual execution of durable directives and proxies. PMID- 7487257 TI - The benefits of estrogen replacement therapy on oral health. The Leisure World cohort. AB - BACKGROUND: Several studies have demonstrated a relationship between tooth loss and alveolar residual ridge resorption and systemic osteoporosis. The recognized benefit of estrogen replacement therapy (ERT) in postmenopausal osteoporosis prompted review of the Leisure World Cohort Study regarding the effects of ERT on tooth loss and the need for dentures in older women. METHODS: The Leisure World Cohort, established in 1981, comprises 13,979 residents (8877 women) of a retirement community. Of the 5935 cohort women alive in 1992, 3921 returned a dental survey with sufficient information to analyze relationships with ERT as reported on the original baseline questionnaire. RESULTS: After adjusting for age, tooth loss and rates of edentia were significantly lower in estrogen users than in nonusers (relative risk for edentia, 0.64; 95% confidence interval, 0.51 to 0.79). The proportion of women with edentia decreased with increasing duration of ERT. Denture wearing was also less common in estrogen users than in nonusers (relative risk, 0.81; 95% confidence interval, 0.71 to 0.93) and decreased with increasing duration of ERT. CONCLUSION: Estrogen replacement therapy may be beneficial in preventing tooth loss and the need for dentures in older women. PMID- 7487258 TI - Reducing low-density lipoprotein cholesterol levels in an ambulatory care system. Results of a multidisciplinary collaborative practice lipid clinic compared with traditional physician-based care. AB - BACKGROUND: The Cincinnati (Ohio) Department of Veterans Affairs Medical Center Lipid Clinic was established as a collaborative practice to treat patients with substantially elevated serum cholesterol levels referred from the General Internal Medicine Clinic. The Lipid Clinic team (led by a clinical nurse), included a clinical pharmacist, nurse practitioner, dietitian, and clinical psychologist. A consultant cardiologist reviewed all laboratory tests and confirmed therapeutic decisions at a weekly preclinic meeting. OBJECTIVE: To compare the success of a limited term of treatment in the Lipid Clinic with that of standard physician-based care in the General Internal Medicine Clinic in achieving the goals recommended by the National Cholesterol Education Program 1 for low-density lipoprotein cholesterol. METHODS: A convenience sample of age matched patients with total cholesterol levels greater than 6.85 mmol/L (265 mg/dL) was selected from each clinic (Lipid Clinic, n = 60; General Internal Medicine Clinic, n = 60). Fasting lipid profiles were drawn in the free-living state and in the sitting position, and matched by month. Treatment of patients in the Lipid Clinic group consisted of evaluation and treatment of secondary causes of hyperlipidemia, goal setting, and treatment according to the National Cholesterol Education Program I algorithm. Counseling and education were individualized. Outcomes were determined after four visits (12 and 18 months for the Lipid Clinic and General Internal Medicine Clinic groups, respectively). Patients in the two groups had comparable risk factors, including presence of coronary heart disease. RESULTS: After four clinic visits, patients in the Lipid Clinic group were four times more likely to reach a National Cholesterol Education Program I goal of a low-density lipoprotein cholesterol level less than 3.36 mmol/L (130 mg/dL) than were comparable patients in the General Internal Medicine Clinic group (relative risk, 4.1; 95% confidence interval, 1.4 to 12.7; P < .001). CONCLUSION: These results support multidisciplinary, goal-oriented collaborative practice as an efficacious model of preventive medicine and health care provision. PMID- 7487259 TI - Preventing pneumococcal bacteremia in patients at risk. Results of a matched case control study. AB - BACKGROUND: Three randomized controlled trials of the effectiveness of pneumococcal vaccine in elderly and chronically ill adults in the United States have failed to show significant protective efficacy during 44,213 person-years of follow-up. Case-control studies have greater statistical power to detect significant prevention of rare diseases such as pneumococcal bacteremia, but they also have a greater susceptibility to bias, necessitating consistent results from multiple studies. Three case-control studies at two different universities have shown prevention of systemic infection, but another study found no benefit. METHODS: Patients with pneumococcal bacteremia who were at least 2 years old and had chronic illness indicating the need for pneumococcal vaccine, or who were at least 65 years old were compared with matched control subjects for frequency of prior vaccination. Matching variables included date of admission, age, sex, race, type and duration of chronic illness serving as the major vaccine indication, number of vaccine indications and number of medical hospitalizations since licensure of the pneumococcal vaccine in 1978, and type of primary medical care. RESULTS: Pneumococcal vaccination was documented in the records of six (7%) of 85 cases and 26 (17%) of 152 control subjects, suggesting 81% efficacy in conditional logistical regression analysis (95% confidence interval, 34% to 94%, P = .008). CONCLUSIONS: Four case-control studies at three universities have now demonstrated significant protective efficacy of pneumococcal vaccine for preventing pneumococcal bacteremia. The development of antibiotic-resistant Streptococcus pneumoniae indicates an urgent need for an increased rate of vaccination among high-risk patients and for the development of more immunogenic conjugate vaccines that may enhance efficacy among elderly and immunocompromised patients as well as infants. PMID- 7487260 TI - Gender differences in device therapy for malignant ventricular arrhythmias. AB - BACKGROUND: Several recent reports have suggested a gender bias in the treatment of cardiovascular disease. It is not clear whether this is true in the treatment of malignant ventricular arrhythmias. OBJECTIVES: To perform a retrospective chart review of 130 patients evaluated for malignant ventricular arrhythmias between July 1990 and June 1992. To compare baseline cardiovascular and clinical parameters and treatment modalities, including cardioverter-defibrillator implantation rates, between women and men. RESULTS: There was no significant difference in the percentage of women and men who were advised to have cardioverter-defibrillator implantation (61% vs 53%) or who underwent cardioverter-defibrillator implantation (46% vs 52%). Women had a lower incidence of coronary artery disease than men (61% vs 85%, P < .01), a lower incidence of myocardial infarction (46% vs 75%, P < .01), and a higher mean left ventricular ejection fraction (38% vs 32%, P = .02). Of patients with indications for cardioverter-defibrillator implantation, significantly more women refused a device than men (19% vs 2%, P = .01), and significantly more women were considered medically ineligible for cardioverter-defibrillator implantation despite having less severe heart disease as a group (12% vs 0%, P = .04). This resulted in significantly fewer women receiving a defibrillator than men with similar indications (18 of 26 women vs 47 of 48 men, P < .01). Of patients who received defibrillators, significantly more women received investigational devices (50%) than men (21%) (P < .05) (35% of women and 19% of men with indication for cardioverter-defibrillator implantation). In-hospital mortality was low in both groups (women, 0%; men, 4%). The 30-month mortality in patients with indications for device intervention was similar in both groups (women, 21%; men, 19%). CONCLUSIONS: No evidence of difference was found between women and men in the rates of recommendation for, or implantation of, implantable cardioverter defibrillators. Women refused device implantation more often than men, and they may be considered medically ineligible for device implantation more than men. This combination results in fewer women with medical indications for cardioverter defibrillator implantation receiving defibrillators than men. This difference does not appear to be associated with increased short-term mortality. PMID- 7487261 TI - Value of tongue biting in the diagnosis of seizures. AB - BACKGROUND: Seizures are rarely witnessed by physicians, and the diagnosis is usually made on the basis of the history. Tongue biting is classically considered to favor a diagnosis of epileptic seizure. The usefulness of tongue biting in the differential diagnosis of seizures was evaluated. METHODS: A prospective study of the presence of oral lacerations in 106 consecutive patients admitted to our Epilepsy Monitoring Unit and a retrospective study of a population of 45 patients with syncope were performed. The relationship between tongue biting and diagnosis (epileptic vs nonepileptic events) was analyzed. RESULTS: Of the 106 monitored patients, 63 had episodes characterized by bilateral motor activity, complete loss of consciousness, or both; 34 patients had epileptic seizures, while 29 patients had exclusively nonepileptic episodes. Eight patients suffered an oral laceration; all involved the side of the tongue, and all had documented epileptic seizures. Of the 45 patients with syncope, in only one was the tongue lacerated, and this was at the tip. Tongue biting had a sensitivity of 24% and a specificity of 99% for the diagnosis of generalized tonic-clonic seizures. Lateral tongue biting was 100% specific to grand mal seizures. CONCLUSION: Tongue biting, particularly if it is lateral, is highly specific to generalized tonic-clonic seizures. PMID- 7487262 TI - Primary pulmonary hypertension and the human immunodeficiency virus. Report of two cases and a review of the literature. AB - We report two cases of human immunodeficiency virus (HIV) seropositivity and pulmonary hypertension seen at our institution and present a comprehensive literature review and available histopathologic findings of the association between HIV seropositivity and pulmonary hypertension. Studies and reviews pertaining to HIV seropositivity and pulmonary hypertension were identified through a MEDLINE search and reference citations. All studies and series found in the MEDLINE search were reviewed and are discussed in this article. Where data were available, comparisons and analyses were made between groups of reported cases of HIV seropositivity and pulmonary hypertension with regard to the following parameters: sex distribution, mode of acquiring HIV infection, presence or absence of the acquired immunodeficiency syndrome, CD4 cell counts, PO2 or oxygen saturation by pulse oximetry, concurrent lower respiratory tract infection, and histopathologic features. We conclude that there is strong evidence for pulmonary hypertension associated with HIV infection that is histologically indistinguishable from primary pulmonary hypertension. Consequently, HIV-seropositive patients with unexplained dyspnea should be evaluated for primary pulmonary hypertension. Prospective studies in HIV-positive patients are indicated. PMID- 7487263 TI - Delirium, DRGs, and documentation. PMID- 7487264 TI - Undying care for terminal patients. PMID- 7487265 TI - [Apropos of patients with inoperable coronary disease]. PMID- 7487266 TI - [Prognosis and predictive factors of mortality in patients with inoperable coronary disease]. AB - In the period between September 1979 and December 1986, 105 out of 2,178 consecutive patients with coronary artery disease (men 93, women 12; age : 58.1 +/- 10 years; previous myocardial infarction : 67%) were considered inoperable because of poor distal coronary circulation (84.8%), left ventricular dysfunction (3.8%) or both (11.4%). Fifty-four clinical, ergometric and angiographic parameters were examined at inclusion. The mean follow-up was 69 +/- 40.2 months (1 to 146 months). The Kaplan Meier 10 year survival rate was 43%. Only 25% of the population remained free of major cardiac events. Multivariate analysis showed that only the coronary angiographic score had a significant predictive value. The authors conclude that the long-term prognosis of these patients is poor. Some did undergo coronary bypass surgery secondarily, and their prognosis was good, suggesting that revascularisation, even if only partial and at high risk, should be considered. PMID- 7487267 TI - [Valvular replacement for aortic stenosis in octogenarians. Apropos of a consecutive series of 200 cases]. AB - From 1978 to 1992, 200 patients aged 80 to 90 (average 82.16 +/- 2.04 years) with calcific aortic stenosis, underwent isolated aortic valve replacement (187 cases, 93.5%), or associated with coronary bypass surgery (12 cases, 6%), or mitral valve replacement (MVR) (1 case, 0.5%), or surgery of the ascending aorta (4 cases, 2%). These 200 patients represented 7.4% of the 2,716 cases of aortic stenosis operated during the same period. One hundred and forty-eight patients (74%) were in NYHA classes III or IV. The operative mortality was 11.5% (23 patients) and the average hospital stay in the surgical department was 12.7 days (4.83%). The follow-up of the 177 patients who were discharged from hospital was complete (100%) with an average of 2.8 years +/- 2.1 (1 month to 10.6 years). Forty-nine secondary fatalities were observed (28%). Of the 128 survivors at the end of the study, 127 (98.6%) were in the NYHA classes I or II. The actuarial 1, 3 and 5 year survivals were 81.7, 74.8 and 57.1%, respectively. The survival curve of the operated patients was identical to subjects of the same age without aortic stenosis. Despite the high but acceptable operative risk due to the age, valvular replacement surgery is justified by the double benefit of increased longivity and improved quality of life. PMID- 7487268 TI - [Patency of left border artery bypass after 1 year; comparison of three techniques]. AB - The one year results of three techniques of bypass grafting of the artery of the left border of the heart were compared in a retrospective study in 120 patients all undergoing left anterior descending bypass grafting with an internal mammary artery. Group I comprised 38 consecutive patients: the left border artery was bypassed with a venous graft. Group II comprised 49 consecutive patients who had the left border artery bypassed by an internal mammary artery in situ. Group III comprised 33 consecutive patients who had the left border artery bypassed by an internal mammary graft issuing from a Y-shaped construction (right internal mammary artery as a free graft reimplanted into the left internal mammary artery). The operative mortality, morbidity and functional results were comparable in the three groups. The rate of angiographic success of the left border artery graft at one year was: 65.7% in group I, 89.5% in group II and 87.8% in group III. The one year patency of internal mammary artery grafting of the artery of the left border was higher than that of venous grafts (p < 0.05). PMID- 7487269 TI - [Practical experience of specific catheter ablation of atrial flutter in 110 patients]. AB - The object of this study was to assess the efficacy and risks of radiofrequency ablation of common atrial flutter and to determine the optimal site of ablation in a large population of patients. Three different methods were used to determine the site of ablation: the first was anatomical and electrophysiological whilst the two others were based essentially on anatomical landmarks for localising the critical zone of the reentry circuit. Recent studies report that radiofrequency ablation is effective in interrupting and preventing recurrences of common atrial flutter both by using anatomical and electrophysiological methods. Nevertheless, a larger series of patients was necessary to establish the efficacy and to determine the optimal site of ablation. A series of 110 consecutive patients with common atrial flutter resistant to antiarrhythmic drugs was studied. The site of ablation of the first 50 patients was determined using both anatomical landmarks and electrophysiological parameters. The anatomical zones were: zone 1, between the septal leaflet of the tricuspid valve and the orifice of the inferior vena cava; zone 2, between the septal leaflet of the tricuspid valve and the ostium of the coronary sinus, and zone 3: between the orifice of the inferior vena cava and the ostium of the coronary sinus. The electrophysiological criterion was an endocavitary auriculogramme occurring during the plateau phase preceding the F wave of the flutter.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7487270 TI - [Importance of atrioventricular synchrony in hypertrophic obstructive cardiomyopathy treated by cardiac pacing]. AB - This study was undertaken to evaluate the effect of permanent dual-chamber cardiac pacing in hypertrophic obstructive cardiomyopathy resistant to medication, paying particular attention to atrioventricular synchrony. Sixteen patients, mean age 59 +/- 13 years (range 36 to 80 years) were divided into two groups after in initial catheter study performed under temporary VDD pacing between March 1990 and April 1993. In group I (n = 11), the gradient was decreased by more than 50% whereas in group II (n = 5), the gradient was unchanged or reduced by less than 50%. The reduction of the gradient was immediately significant in group I, the mean value falling from 104 +/- 33 mmHg (range 60 to 170 mmHg) to 25 +/- 13 mmHg (range 10 to 60 mmHg) (p < 0.0001). In group II, the gradient only decreased initially from 132 +/- 13 mmHg (range 120 to 150 mmHg) to 88 +/- 25 mmHg (range 50 to 130 mmHg) (p < 0.003) but improved atrioventricular synchrony, obtained secondarily either by pharmacological prolongation of the PR interval (association of betablocker and verapamil) or by ablation of the atrioventricular junction, improved the haemodynamic benefits. The residual gradient recorded on the 7th day was only 26 +/- 15 mmHg (range 10 to 50 mmHg) (p < 0.0001). The comparison of the two populations showed that the mean PR interval was shorter in group II (p < 0.016) and the mean value of the optimal AV Delay (the longest AV Delay with complete ventricular capture) was also lower (p = 0.001).(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7487271 TI - [Transluminal coronary angioplasty in patients with left ventricular dysfunction; immediate and long term results]. AB - Transluminal coronary angioplasty is increasingly performed in patients with left ventricular dysfunction. However, there is little data available concerning the outcome of these patients. The aim of this retrospective study was to assess the immediate and long-term results in 90 patients (76 men and 14 women) with left ventricular ejection fractions < or = 35% (average 29 +/- 5%) undergoing angioplasty between 1980 and December 1992. Eighty-seven patients (96%) had a history of infarction, 27 (30%) had already at least one episode of left ventricular failure and 34 (38%) had unstable angina at the time of angioplasty. The coronary disease was usually multi-vessel. A total of 118 lesions were dilated with a primary success rate of 77% (91/118): 86% (82/95) in non-occlusive stenoses and 39% (9/23) in complete obstructions. The total success rate per procedure was 72% (65/90) with a hospital mortality rate of 5.5% (5 cases). The mean follow-up period was 53 +/- 47 months. Twelve patients died during this period and two were lost to follow-up. The total and cardiovascular mortality at the end of the study was 19% (17/88). Eight of the 71 survivors underwent another revascularisation procedure (4 bypasses and 4 angioplasties); 21 (29%) have stable angina, 50 (71%) have no anginal pain and 61 (86%) have antianginal treatment. The probability of survival at one and four years was 81 +/- 4% and 79 +/- 5%, respectively. A multivariate analysis using the Cox model showed three independent prognostic factors for long-term mortality: triple coronary vessel disease, the best predictive factor, left ventricular ejection fraction and female gender.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7487272 TI - [Radiofrequency ablation of intranodal tachycardia]. AB - The first radical approach to the treatment of atrioventricular nodal reentrant tachycardia was surgical dissection of the perinodal region. This technique has been replaced by the delivery of radiofrequency energy by an ablation catheter to the region of the atrioventricular node. The aim of this report is to describe the authors' experience of atrioventricular nodal application of radiofrequency current. The study comprised 53 cases (32 women and 21 men, mean age 46 +/- 17 years) with frequent attacks of reciprocating tachycardia. Endocavitary electrophysiological investigation confirmed the intranodal reentrant mechanism. The region of application of the radiofrequency current was located radiologically and then the precise site determined by the recording of nodal electrical activity. The appearance of junctional rhythm during the procedure was also used as a means of identification of the zone of ablation. Dual conduction persisted after ablation in 35 patients. However, no episode of tachycardia could be induced after the procedure. The AH interval increased during application of radiofrequency current in 3 cases but this abnormality regressed in the 2 months following the procedure. Recurrences of nodal reentrant tachycardia were observed in 14 cases (26%), 24 hours to 2 months after ablation. The rate of recurrence was significantly higher in patients who did not have a junctional rhythm during application of the radiofrequency current (62% vs 4%, p < 0.05). The number of recurrences was also greater in the group of patients with persistence of slow intranodal conduction after the radiofrequency ablation (p < 0.04). A second session of radiofrequency ablation was undertaken in 14 patients and a third session was required for 2 of them.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7487273 TI - [Septicemia and endocarditis related to transvenous pacing leads of pacemakers: surgical indications and results]. AB - Endocarditis of transvenous pacing leads is a rare condition. The authors review a series of 15 patients who developed bacteriologically proven septicaemia and/or endocarditis related to transvenous pacing leads, operated between 1988 and 1993. The interval between the last manipulation of the pacemaker and the onset of endocarditis was about 6 months. Six patients had had haematoma and/or infection of the pacemaker site. Endocarditis presented with chronic pyrexia (14 cases) associated with septicaemia (6 cases) and chronic local suppuration (1 case). The interval between the beginning of the pyrexia and the diagnosis was 3.4 months. Echocardiography showed a mass attached to the pacing lead in 8 cases and tricuspid valve vegetations in 4 cases. Blood cultures were positive in 13 patients and local wound swabs identified the organism in 1 patient. The commonest causal agent was the staphylococcus (epidermis in 7 cases, aureus in 4 cases). Appropriate antibiotic therapy was only effective in 1 case. The surgical indication in 13 cases was persistence of infection associated with pulmonary embolism (3) or tricuspid regurgitation (2). Complete ablation of the prosthetic material was performed by a peripheral vascular approach (2 cases), by a right atrial approach (1 case) and under cardiopulmonary bypass in 12 cases. The peroperative findings were of tricuspid valve vegetations (4 cases), thrombi on the pacing lead (7 cases) or in the right heart chambers (2 cases) or pulmonary artery (2 cases). The associated procedures performed under cardiopulmonary bypass were tricuspid valve repair (2 cases) and pulmonary thrombectomy (2 cases). Temporary and permanent epicardial leads were implanted in 10 patients.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7487274 TI - [Reflections of an expert committee of the French Society of Cardiology on the use of methylergometrine maleate (Methergin) for the detection of abnormal coronary vasomotricity]. PMID- 7487275 TI - [Thrombolysis after 75 years of age: benefits, risks, indications]. AB - Hospital mortality of myocardial infarction in patients over 75 years of age ranges from 25 to 33% without thrombolysis. Large scale trials of intravenous thrombolytic therapy including elderly patients showed that age itself is not a contra-indication to thrombolytic therapy. However, on the one hand, contra indications are more common, and, on the other, the characteristics of infarction in the elderly are unsuitable so that thrombolysis is rarely used: in the MITI study, only 15% of over 75 years old patients would have benefitted from this therapy. The benefits of this treatment in patients over 75 years of age have been demonstrated: they were significant in Collin's meta-analysis (hospital mortality 24.9 vs 28.8%). In a more recent analysis (FFT study) the benefits of thrombolytic therapy after 75 years of age were shown providing the delay of inclusion (< 6 hours) and classical electrocardiographic criteria (ST elevation) were respected. Moreover, age is a risk factor for cerebral haemorrhage (prevalence 2.08% in patients over 75 years of age in the GUSTO study). The results of this study showed that, despite the development of thrombolysis, age was the main risk factor for hospital mortality and was independent of other risk factors. Therefore, the indication of intravenous thrombolytic therapy in elderly patients should be considered with respect to the characteristics of infarction (delay, site, presence of subendocardial ischaemia), the contra-indication related to potential bleeding and the possibilities of performing emergency angioplasty. PMID- 7487276 TI - [Conceptual basis and diagnosis of primary hyperaldosteronism]. AB - Primary aldosteronism comprises two different conditions, the tumoral form usually caused by an adenoma and the idiopathic form due to nodular hyperplasia of the two adenals. More rarely, an adenoma of the adrenal cortex, glucocorticosteroid-sensitive hyperplasia, and angiotensin-sensitive adenoma or an autonomous nodule transformed to primary tumoral hyperplasia, may be observed. Primary alderosteronism may be conceived as a spectrum of genetic abnormalities which express themselves either by hyperplasia or by a tumour. A defect in steroid genesis and prolonged stimulation of the cortex would lead to the formation of nodules which may become autonomous and generate a tumour. Hypertension may be isolated. Detection requires three sampling of serum potassium in all hypertensive patients, a study of the aldosterone-renin axis when the value is less than 3.6 mEq, or whenever the hypertension is severe or resistant to treatment. The diagnosis is made by the association of an increased plasma aldosterone level before getting up in the morning and a plasma renin unaffected by orthostatism. The choice of medical or surgical treatment depends on the uni- or bilateral anatomic substrate. Computerised tomography, very sensitive but not specific, like hormonal studies, often provides incomplete answers. Adrenalectomy is indicated in the presence of a mass of centimetric proportions with concordant results of the dynamic test. In other situations, investigations are continued with the search for an aldosterone gradient by selective venous sampling. This is very valuable to determine the lateralisation but fails in 25% of cases, and its results have to be compared with those of imaging techniques: CT scan, venography and, when necessary, scintigraphy.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7487277 TI - [Thrombolytic therapy with rt-PA for thrombosis of tricuspid valve prosthesis during pregnancy]. AB - The authors report the case of a woman with thrombosis of a tricuspid Saint Jude prosthesis during the fourth month of pregnancy. A first course of thrombolytic therapy with rt-PA reestablished normal prosthetic valve function but was followed by a threatened abortion and severe uterine haemorrhage. An early rethrombosis of the prosthetic valve led to interruption of the pregnancy after failure of a second course of thrombolysis, and to replacement of the tricuspid valve prosthesis. This case illustrates the problems of pregnancy in women with mechanical cardiac prosthetic valves and the difficulties of treatment. PMID- 7487279 TI - [Struma cordis: right intraventricular ectopic thyroid]. AB - The authors report the case of a benign tumour composed of hyperplasic thyroid tissue in the right ventricle, diagnosed in a 43 year old woman by echocardiography after a syncopal episode. The outcome was favourable with a 13 year follow-up after surgery. PMID- 7487278 TI - [Transesophageal echocardiographic diagnosis of left ventricular parietal rupture, a complication of myocardial infarction]. AB - The authors report the case of postero-lateral myocardial infarction complicated by free wall rupture at the 48th hour. The diagnosis was suspected clinically and transoesophageal echocardiography showed the appearance of pericardial effusion in a patient difficult to examine by conventional echocardiography. Transoesophageal echocardiography was performed and well tolerated. The transgastric view showed a thrombus adjacent to a thinned and lacerated postero lateral wall. The clinical outcome was favourable after surgical repair. PMID- 7487281 TI - [Thrombolytic treatment of pulmonary embolism. The end of armament race?]. PMID- 7487280 TI - [History of the first radiofrequency ablation]. PMID- 7487282 TI - [Use of radionuclide isotopes by cardiologists. Results of a French survey. Study group: "Nuclear Cardiology" of the French Society of Cardiology and the Society of Nuclear Medicine and Biophysics]. AB - The authors report the results of an enquiry carried out amongst French cardiologists to determine their opinion about radionuclide investigations in cardiological practice. Of the 5,050 cardiologists contacted, 1,431 (28.3%) replied. The cardiologists were attracted by the non-invasive nature (84%) of radionuclide techniques and their complementarity with other methods of investigation (74%) but regretted their cost (55%), their lick of availability in emergencies (35%) and, in general, their difficult of access (30%). Only 38% of the cardiologists who replied used radionuclide investigation on an everyday basis, this being impossible for some because of the distance to the nearest centre with these facilities (on average 32 km but exceeding 50 km in 29% of cases, and the delay before obtaining an appointment (average 13 days) which was often excessive, especially for myocardial scintigraphy (thallium or equivalent). Each cardiologist prescribed an average of 5 myocardial scintigraphies, 3 pulmonary scintigraphies and 2 radioisotopic ventriculography per month. These results seem to be an overestimation; in fact, radionuclide investigations are relatively underemployed, probably more because of the factors cited above than because of lack interest or quality, these latter two points being generally judged positively by the majority of cardiologists. Cardiologists require more from the conclusions of the investigation than a simple description of the images obtained. As with all other investigations that they perform themselves, the interpretation of the results must take the clinical context into consideration and form part of the diagnostic and therapeutic management of the patient.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7487283 TI - [Management of myocardial infarction in the Rhone-Alps area. Are there many variations in practice?]. AB - The aim of this study was to determine the diagnostic and therapeutic strategies after myocardial infarction and to examine variations in medical and surgical practice with respect to the severity of disease, status of the hospital and patients' characteristics. The method used was a prospective study with follow-up at 30 days and 18 months. The subjects came from an exhaustive cohort of all patients admitted to hospital for myocardial infarction during the month of April 1991 in 57 public and private hospitals in the Rhone-Alps region (n = 311). The patients were identified after admission by consulting physicians of the Department of Social Security. The study included all patients with acute myocardial infarction with at least two of the three usual diagnostic criteria (prolonged, constrictive chest pain, enzyme increases and electrocardiographic changes). The study excluded patients who were dead before arrival at hospital. The parameters analysed included the clinical management, use of echocardiography, exercise stress testing, myocardial scintigraphy, coronary angiography, thrombolysis, angioplasty and coronary bypass surgery in the first 30 days after admission. The severity of infarction was assessed by seven clinical, enzymatic and electrocardiographic criteria by physicians from the Department of Social Security (pain, syncope, shock, left ventricular dysfunction, elevation of CPK > 1000 IU, anterior or extensive necrosis, arrhythmias). The mortality rate of this cohort was calculated from hospital statistics and then by enquiring in the town halls of the region. The demographic features of the cohort were marked by a predominance of men (69.5%) and a relatively high mean age (69 years; 23% over 80 years). Complementary investigations were used with the following frequencies: echocardiography, 61.1%; coronary angiography, 26.4%; exercise stress testing 22.8%; myocardial scintigraphy, 5.5%.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7487285 TI - [Valve ring abscesses: apropos of 59 cases. A multicenter study]. AB - The authors report the results of a multicenter study which recensed 59 cases (46 men, 13 women, average age 59.8 +/- 14 years) of valve ring abscesses defined by echocardiographic criteria alone (20 cases) or by echocardiographic and/or operative criteria (39 cases). The site of abscess was aortic (53 cases), mitral (5 cases) and tricuspid (1 case). The abscess complicated a prosthetic valve in 34 cases, occurred with a non-pathological valve in 11 cases or a pathological valve in 14 cases. Taking the 39 operated patients as a reference, the diagnostic sensitivity of transthoracic echocardiography was 25% and that of transoesophageal echocardiography: 88%. Bacterial vegetations were diagnosed with a sensitivity of 55% for transthoracic and 88% for transoesophageal echocardiography. The mediocre results of transthoracic echocardiography make transoesophageal echocardiography mandatory when there is a clinical suspicion of abscess: transoesophageal echocardiography should be systematic in prosthetic valve endocarditis and widely employed in native aortic valve endocarditis. The clinical outcome of these cases was: 39 cases were operated: global mortality of 23%, and 18 cases were treated medically, surgery having been declined for various reasons: old age (2 patients), operative risk (1 patient), patient refusal (4 patients), general condition considered to be too good to justify surgery (11 cases, including 6 sterilised abscesses diagnosed some time after the acute infectious phase). The outcome of these 18 patients, who form the biggest non-operated series of valve ring abscesses to date, was studied in detail: 4 died (18% mortality), 1 was operated secondarily for progressive valve dehiscence and 13 had a favourable outcome with an average follow-up period of 2 years.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7487284 TI - [Systematic transesophageal echocardiography during the postoperative first 24 hours after mitral valve replacement]. AB - Systematic transoesophageal echocardiography after the 10th day of mitral valve replacement with a mechanical prosthesis has enable diagnosis of several abnormal conditions: thrombi, strands and paravalvular leaks. The aim of this prospective study was to determine the prevalence of these conditions by biplane transoesophageal echocardiography in the first 24 postoperative hours. Transthoracic and transoesophageal echocardiography was performed on average 12 +/- 3 hours after coming out of the surgical block in 77 consecutive patients who underwent mitral valve replacement with a mechanical prosthesis. Nine patients (11.7%) had appearances of thrombi or strands in the left atrium or auricle. These small thrombi (1 to 1.5 cm2) were not obstructive. No embolic events were observed in the first month of these patients. Spontaneous contrast was seen in the left atrium of 31 patients (40%). The factors associated with the presence of thrombus or strands were advanced age (p = 0.02), presence of spontaneous contrast (p = 0.02) and more dilated left atrium (p = NS) Paraprosthetic leaks were seen in 11 cases (14.3%). In 10 cases, the regurgitant jets were narrow at their origin with little extension into the left atrium. Only one patient had severe regurgitation associated with raised transprosthetic pressure gradients. Cases with paravalvular leaks had a lower incidence of spontaneous contrast (3.2% vs 21.7%) and no thrombosis. Transoesophageal echocardiography demonstrated spontaneous contrast, thrombi and strands in the initial hours following implantation of a mechanical mitral valve prosthesis. The prevalence of these appearances, comparable to that of the results reported with later investigations, underlines the importance of effective anticoagulation from the fist postoperative hours. PMID- 7487286 TI - [Subcutaneous recombinant hirudin in the treatment of deep venous thrombosis. A pharmacokinetic study]. AB - Recombinant hirudin (HBW 023) has a pure and specific antithrombotic activity. It could be more effective than heparin in the treatment of deep venous thrombosis. Its half life is about three hours when administered intravenously which requires continuous infusion whereas subcutaneous administration can ensure stable plasma concentrations and antithrombotic activity over a period of approximatively 12 hours. The aim of the study was to check the safety and clinical and radiographic efficacy of recombinant hirudin administered subcutaneously to patients with recent deep venous thrombosis and to analyse the pharmacokinetics of the product and its effects on tests of coagulation. Ten patients were treated with 0.75 mg/kg of subcutaneous recombinant hirudin twice a day for 5 days. Anticoagulation was performed with standard heparin and acenocoumarol. Bilateral phlebography, pulmonary angiography or ventilation and perfusion scintigraphy were carried out before and on the 5th day of recombinant hirudin treatment. The activated cephalin time and standard anticoagulant tests and the plasma kinetics of recombinant hirudin were assayed between the 1st and 12th hour on the first and fifth days of treatment. The clinical course was simple in all but one patient who had a recurrence of pulmonary embolism on the 4th day justifying thrombolytic treatment. No haemorrhagic complications or secondary biological effects were observed. On the 5th day, control phlebography was unchanged or improved in all patients. The peak plasma concentration of recombinant hirudin was observed between the 3rd and the 4th hour following subcutaneous injection. The activated cephalin time was increased in parallel with increased concentrations of recombinant hirudin.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7487287 TI - [Delayed angioplasty of residual coronary stenosis after thrombolysis of myocardial infarction: prognostic factors of left ventricular function]. AB - The aim of this study was to analyse the prognostic factors of long term left ventricular function after successful delayed angioplasty of residual coronary stenosis after thrombolysis for myocardial infarction. The left ventricular function of fifty patients, aged 54 +/- 10 years, who underwent intravenous thrombolysis 2.9 +/- 1.3 hours after the onset of myocardial infarction (28 anterior, 22 inferior wall) was assessed by angiography at the 6.1 +/- 1.7th day and 6.1 +/- 2.5 months after successful angioplasty undertaken on the 8.3 +/- 5.2nd day. In cases without reocclusion (n = 44), the ejection fraction and the wall motion of the infarcted region improved significantly from 56.8 +/- 12.2% to 59.5 +/- 15.4% (p < 0.002) and from 8.4 +/- 14.8% to 18.3 +/- 24.3% (p = 0.0001), respectively. These parameters did not change in cases with reocclusion (n = 6). When the dilated artery remained patent, an improvement in motion of the infarcted region was observed in 69.5% of hypokinesis (n = 23), 50% of akinesis (n = 10) and 9.1% of dyskinesis (n = 11) (p < 0.005). The improvement in motion of the infarcted zone was negatively correlated with the delay of thrombolysis (r = 0.36; p < 0.02) and peak CPK levels (r = 0.31; p < 0.04). Over 60% restenosis on digitalised densitometry (n = 11) did not compromise the possibilities of improvement, the evolution of wall motion of the infarcted zone and volumes with respect to cases without restenosis (n = 33).(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7487288 TI - [Morphological study by transesophageal echocardiography and clinical aspects of ruptured chordae tendineae in the elderly]. AB - Ruptured mitral chordae tendinae is a classical complication of myxomatous mitral valves or Barlow's syndrome. This complication is controversial in non-myxomatous mitral valve. Of 91 consecutive patients with mitral valve prolapse examined over an 18 months period by transthoracic and transesophageal echocardiography, 42 (18 women and 24 men) with an average age of 76 +/- 8 years (60-93 years) had ruptured mitral chordae tendinae. The thickness of the anterior mitral leaflet measured at the distal third of the valve by transesophageal echocardiography enabled the identification of two groups of patients; group I: > 3 mm (24 patients), average 4.8 +/- 0.8 mm and group II: < or = 3 mm (18 patients), average 2.6 +/- 0.3 mm. The diameter of the mitral ring and left atrium, the length of the anterior mitral leaflet, the left ventricular end diastolic dimensions and fractional shortening, were measured by transthoracic 2D echocardiography (mitral ring) and M mode (other parameters). Ruptured chordae were detected in only 13 cases (31%) by transthoracic echocardiography; 38% were asymptomatic and a chance finding at transesophageal echocardiography. No significant difference was observed between the two groups with respect to age, gender presence of hypertension, dimensions of the cardiac chambers, fractional shortening or localisation of the prolapse related to the ruptured chordae. Fifty eight per cent of patients in group I were in NYHA functional classes 3-4 as compared to 16% in group II (p < 0.02). The size of the left atrium was significantly greater in group I, 51 +/- 8 mm vs 38 +/- 7 mm (p < 0.001).(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7487289 TI - [Is the transesophageal approach preferable to endocavitary approach in the evaluation of Wolff-Parkinson-White syndrome?]. AB - Now that the radical treatment of the Wolff-Parkinson-White syndrome is established, it is essential to evaluate the prognosis of this condition accurately. Initiation of atrial fibrillation is one of the factors which influence the prognosis. The aim of this study was to compare the results of electrophysiological studies performed by the endocavitary and transoesophageal approaches in the measurement of the initiation of atrial fibrillation. Twenty six patients with a patent Wolff-Parkinson-White syndrome were studied by the two methods with a similar protocol: incremental atrial pacing to the Wenckebach point, programmed atrial stimulation using up to two extrastimuli, repeated with an infusion of 20 to 30 ug of isoproterenol. Sixteen patients had reciprocating nodal tachycardia or were asymptomatic (group I) and the other 10 had spontaneous atrial fibrillation (group II). In group I, atrial fibrillation was induced in 9 cases (56%) by the endocavitary and in two cases (12.5%) by the transoesophageal method. In group II, spontaneous atrial fibrillation was reproduced in all cases by the endocavitary and transoesophageal protocols. None of the patients in group I developed atrial fibrillation during follow-up (average 2 years +/- 9 months). The authors observe that all spontaneous atrial fibrillation of the Wolff Parkinson-White syndrome can be triggered by oesophageal stimulation. The prevalence of atrial fibrillation was overestimated by endocavitary studies in asymptomatic or paucisymptomatic patients. The assessment of atrial vulnerability of a Wolff-Parkinson-White syndrome may therefore be performed by transoesophageal electro-physiological studies. PMID- 7487290 TI - [Fibrinolytic inhibitors and prevention of bleeding in cardiac valve surgery. Comparison of tranexamic acid and high dose aprotinin]. AB - In order to assess the effects of tranexamic acid in comparison to the high dose regimen of aprotinin recommended by Royston and considered to be the reference in postoperative bleeding in cardiac surgery, 35 consecutive patients were randomised to two groups according to the product prescribed. The global postoperative bleeding was comparable in the two groups (p = 0.49). One surgical reoperation for haemostasis was required in the reference group. There was one case of renal failure in the same group due to cardiac failure. No thrombotic complications were observed. Platelet function, as judged by the bleeding time and platelet aggregation to ristocetin, was the same in the two groups. The D dimers remained low in both groups, reflecting the absence of intravascular coagulation and fibrinolysis. Tranexamic acid was as effective and as safe as high dose aprotinin. These two substances, in addition to their fibrinolytic inhibitory activity, conserved platelet protection by blocking the action of plasmin. These results seem to justify the preventive use of tranexamic acid from the moment of skin incision, especially in reoperation. PMID- 7487291 TI - [Synchronized anterograde perfusion during percutaneous transluminal coronary angioplasty: preliminary clinical study]. AB - A new circulatory system, "physiologic anteroperfusion system", has been developed and tested in 6 patients with significant proximal coronary artery stenosis. Prolonged and safe balloon inflation was possible without any ischemic signs. The system consists of an electronic cardiac synchroperfusor which, by activating a pulsatile unit, permits increased diastolic anteroperfusion of autologous blood under physiologic pressure through low-profile standard angioplasty catheters. This study reports the results obtained in 6 patients during proximal prolonged percutaneous transluminal coronary angioplasty. Four men and two women suffering from severe exertional angina pectoris, with normal resting left ventricular function, no collaterals and excellent apical two dimensional four-chamber echocardiographic views were studied. After a 90 +/- 10 seconds of control occlusion under continuous monitoring of hemodynamics, electrocardiograms (3 to 4 leads), two-dimensional echo and chest pain grading, a second balloon inflation protected by the physiologic anteroperfusion system at a flow rate of 44 +/- 12 ml/min was performed for fifteen minutes. The ischemic signs present in the myocardium depending on the occluded artery were totally abolished during prolonged inflation protected by physiologic anteroperfusion system. All the patients were successfully dilated and were discharged from hospital the following morning without cardiac enzyme elevation or signs of central or peripheral hemolysis. Conclusion, in 6 patients with severe proximal coronary artery stenosis, safe prolonged proximal angioplasty without signs of ischemia was performed using a new simple physiologic anteroperfusion system, which allows active diastolic flow-pressure controlled autologous arterial blood perfusion, through standard low profile catheters. PMID- 7487292 TI - [Effect of antisense oligonucleotides on myo-intimal hyperplasia in a model of abdominal aortic injury in the rat]. AB - Restenosis at a rate > 30% at 6 months is the major complication of both coronary and peripheral arterial angioplasty. Restenosis is mainly due to proliferation of smooth muscle cells, extracellular matrix and collagen which form a neointima. The proto-oncogene c-myb is a gene with an immediate response which has been implicated in the proliferation and alteration of the phenotype of smooth muscle cells. The antisenses are molecules of single-helix DNA the sequence of which is inverse to that of messenger RNA of the target proto-oncogene. They therefore have the possibility of forming a double helix with the messenger RNA and of preventing its translation. The antisenses of c-myb have already been successfully tested in in vitro and in vivo models of neointimal proliferation. The aim of this study was to demonstrate the efficacy of c-myb antisenses on the proliferation of smooth muscle cells in a model of abdominal aortic injury in the rat. Thirty-five male Wistar rats with an average weight of 350 grams were operated. Smooth muscle cell proliferation was obtained by desendothelialisation of the abdominal aorta from the level of the left renal vein to the aortic bifurcation. Using a randomised, double-blind protocol, 17 rats were given 500 microliters of pluronic gel (control group), 9 a sense oligonucleotide of c-myb in 500 microliters of pluronic gel (sense group) and 9 a c-myb antisense oligonucleide in 500 microliters of pluronic gel (antisense group). Two rats were given fluorescinlabelled antisenses; one was sacrificed 4 hours and the other 24 hours later.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7487293 TI - [Right atrial thrombosis; an unusual late complication of the Fontan procedure. Apropos of a case]. AB - The authors report the case of a large right atrial thrombosis causing acute cardiac failure in an 18 year old patient with tricuspid atresia who had undergone 3 operations: Waterston shunt at 2 months of age, Fontan procedure at 3 years of age with reoperation at 13 years of age. The diagnosis was made at echocardiography and angiography. Surgical management comprised ablation of the thrombus and a tunneling of the right atrium between the inferior vena cava and the atriopulmonary conduit. The immediate postoperative course was complicated by a slowly regressive neurological deficit. The medium-term outcome (one year) is satisfactory with antiarrhythmic and anticoagulant therapy. A review of the literature showed that these thromboses are not exceptional in the early postoperative period for a variety of reasons. Secondary thrombosis is often related to arrhythmias, thus requiring clinical, electrocardiographic and echocardiographic follow-up of these patients and the prescription of antiarrhythmic and anticoagulant therapy. The diagnostic methods are discussed. Echocardiography may be sufficiently explicit to make potentially risky angiography unnecessary. PMID- 7487294 TI - [Aortic dissection and pregnancy. Apropos of a case]. AB - Aortic dissection is a serious disease which rarely affects young women. In this context, it occurs in nearly one out of two cases during pregnancy, usually during the third term. The authors report acute dissection of the ascending aorta (de Bakey type 2) during pregnancy for which rapid cardiothoracic surgical management as a semi-emergency resulted in a favourable outcome for mother and child. PMID- 7487295 TI - [Cardiac tuberculoma; value of echocardiography and therapeutic management apropos of a case]. AB - The authors report the case of a patient with a large mass in the right ventricle which was a tuberculoma without pulmonary disease. The severity of the right ventricular obstruction required surgical intervention with quadri-antitubercular therapy. Myocardial tuberculomas are very rare and usually reported as post mortem findings. Only four cases resulting in cure have been previously reported. Current means of investigation such as echocardiography and endomyocardial biopsy allow rapid diagnosis of these tumours and should lead to better medical management with possible surgical intervention and a higher therapeutic success rate. PMID- 7487296 TI - [Percutaneous dilatation of subaortic stenosis in an adult: a 3-year follow-up]. AB - Subaortic stenosis is usually diagnosed in the first years of life and treated surgically. The authors report the case of stenosis by a subaortic membrane diagnosed in an adult, treated by percutaneous balloon dilatation with a satisfactory outcome at 3 years. PMID- 7487297 TI - [Variability of sinus rhythm and cardiac failure]. AB - Under pathological conditions, the autonomic nervous system, which continually regulates cardiac function, may have adverse effects on pump function and electrical activation. The sinus rhythm is an indirect marker of the different components of the autonomic nervous system and changes in the heart rate reflect the permanent variation of vagal and sympathetic tone. Analysis of variability in the temporal or frequential domaines, provides information about the modulation of the different components of the autonomic nervous system. In the post infarction period, studies have shown that the heart rate variability is an independent predictive factor of cardiovascular mortality. The data available in patients with cardiac failure is limited. There is a decrease in heart rate variability and this decrease seems to be associated with a higher risk. PMID- 7487298 TI - [Baroreflexes and congestive heart failure]. AB - Abnormal responses are found in the early stages of heart failure with increased sympathetic and decreased parasympathetic activity, causing peripheral arteriolar vasconstriction and tachycardia respectively. The cardiopulmonary baroreflex may be studied by decreasing venous return ("low body negative pressure") and by measuring vascular resistance forearm. The arterial baroreflex may be studied by changing aortic pressures (by intravenous phenylephrine or nitroglycerin). Orthostatism and the tilt test deactivate the cardiopulmonary and arterial baroreflexes simultaneously. These baroreflexes are impaired in patients with heart failure. Their activation does not cause the usual sympatho-inhibition so contributing to increased sympathetic tone. This dysfunction may result from a change at any point on the reflex pathway: the baroreceptors themselves, the afferent, central and efferent pathways. It is selective as during the cold pressor test, the vasoconstrictor response remains intact. One of the possible mechanisms of baroreflex dysfunction in heart failure is loss of sensitivities of the baroreceptors. This may be multifactorial: structural abnormalities, changes in compliance or functional abnormality. Even if the loss of sensitivity is partially related to a change in compliance, other factors play a role. It is more functional than structural abnormalities because, after cardiac transplantation, the baroreceptors regain their sensitivity within 2 to 3 weeks. Excessive Na-K dependent ATPase activation of the smooth muscle cells of the carotid sinus could lead to hyperpolarization of the cell membrane, so reducing the excitability of the receptor. Aldosterone is one of the factors which could activate the Na-K ATPase, as this hormone directly increases pump activity and favorizes the synthesis of new pumps in the vascular smooth muscle cells.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7487299 TI - [Value and limitations of methods for measuring left ventricular mass]. AB - M-mode echocardiographic measurement of the left ventricular mass is inaccurate when the hypertrophy is asymmetric and the ventricule very deformed. The routine calculation of the mass is based on several hypotheses, verified in normal and hypertensive subjects: the standard error (SEE) is 30-40 g, 10-15% (r > 0.9). The standard deviation of inter-examination differences (> 25 +/- 30 g with a variation coefficient c = 10-15%) makes it difficult to appreciate variations of mass in a given patient. Two-dimensional echocardiographic measurement of left ventricular mass requires the use of geometric formulae which have not been validated in the cardiomyopathies. The absence of a consensus on the models used has favorized the use of MRI and of ultrafast computed tomography. MRI measurement of mass has been validated in normal and ischaemic hearts (r > 0.97, SEE < 8 g, c = 15%) but the times of acquisition are long. Using ultrafast CT, not universally available, this measurement has been validated in vivo and in vitro, including in cardiomyopathy for which the SEE is low (6%) and reproducibility excellent (c = 4-8%), comparable with results in normal subjects. PMID- 7487300 TI - [Clinico-pathological polymorphism of hypertrophic cardiomyopathy in echocardiography]. AB - An echocardiographic Study of 322 adults (age: 40 +/- 16 years), belonging to 20 families with hypertrophic cardiomyopathy (HCM), was undertaken. Affected subjects had a LV diastolic wall thickness > 13 mm. The patients were classified according to the distribution of left ventricular hypertrophy (LVH) and by Maron's classification: 189 subjects were normal, 127 were affected and 6 could not been classified. By Maron's classification: 3% were type I, 33% were type II, 58% were type III and 6% were type IV. LVH was asymmetrical in 95% of cases (septum/posterior wall ratio > 1.3). The familial distribution of LVH of the 4 families in which HCM was genetically related to different loci (chromosome 11, 14 exon 13, 14 exon 8, fifth locus); the LVH was analysed from two short axis LV parasternal views and each plane was divided into 5 segments. The distribution of LVH was said to be identical between two first degree relations when all the same segments were affected, similar when they differed by only 1 or 2 segments and different when they differed by 3 or more segments. In the 26 pairs studied, LVH was identical in 2/26 (8%), similar in 11/26 (42%) and different in 13/26 (50%). Familial HCM usually gives rise to asymmetrical LVH affecting the septum and free wall. An identical distribution in 50% of affected first degree relatives. PMID- 7487301 TI - [Hypertrophic cardiomyopathy: practical application of genetic research]. AB - Hypertrophic cardiomyopathy is usually familial with an autosomal dominant mode of transmission. The condition is genetically heterogeneous. The first defective gene to be described was that of the beta heavy chain of cardiac myosin (beta MHC, chromosome 14 q11-q12) where over 20 different localised false sense mutations and a hot point of mutation in exon 13 have been reported. This locus seems to be implicated in 30% of families studied. Systematic screening of the genome has led to the detection of new sites on chromosomes 1, 11 and 15. No gene has yet been identified in these three new loci. For all new families, present strategy consists in determining the locus responsible for the disease by linkage analysis. For beta-MHC locus a panel of several markers (the two MYO I and MYO II microsatellites of the beta-MHC gene and two new AFN microsatellites) has been established which allows accurate detection of whether the haplotype cosegregates with the disease, even in relatively small families. It is then necessary to define the mutation by PCR amplification of the exons followed by electrophoresis on Hydrolink-MDE gels and sequencing of the mutant exons. Some subjects are genotypically affected but phenotypically normal and it seems the penetrance varies with the type of mutation. Analysis of the genotype/phenotype relationship should be continued in order to improve our knowledge of the consequences of each mutation. The genotype diagnosis is therefore complex and cannot be undertaken at present of the examination of the propositus. Whole families must be sampled.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7487302 TI - [Hypertrophic cardiomyopathy: place and limitations of medical therapy]. AB - The development of echocardiography and kindred studies have enabled the detection of an increasing number of asymptomatic forms of hypertrophic cardiomyopathy. Drug therapy, by far the commonest therapeutic method of managing this condition, has only been shown to be effective on the symptoms. Therefore, due to the risk of iatrogenic disease, medication is only proposed to symptomatic patients: high dose betablockers is the usual treatment; when ineffective or contra-indicated, verapamil may be prescribed at progressively increasing doses under strict medical control. These products may need to be given at high doses in order to be effective but excessive bradycardia may be a limiting factor. Amiodarone alone or in association, may be preferred in forms with arrhythmias. The comparative efficacy of the different drugs, especially in preventing sudden death, requires well designed, controlled, therapeutic trials. Their results, if positive, would question the dogma of only treating symptomatic patients. PMID- 7487303 TI - [Value of cardiac pacing in hypertrophic obstructive cardiomyopathy refractory to medical treatment]. AB - Recent studies have shown that dual-chamber cardiac pacing could improve patients with hypertrophic obstructive cardiomyopathy resistant to medical treatment. After a brief historical review, the authors report the present status of this original therapeutic option. The results of different studies are in agreement and show that the functional improvement is associated with a large reduction of the intraventricular pressure gradient with permanent DDD pacing. Although the exact mechanism of the improvement remains unknown, all groups underline the importance of complete and permanent ventricular capture. This requires precise regulation of the atrioventricular delay which in most cases must be programmed at a value inferior to 90 ms, which compromises ventricular filling which is already abnormal under basal conditions. The inefficacy of pacing in some cases is explained by the necessity of selecting a very short atrioventricular delay which alters diastolic function and finally masks the benefits of ventricular capture. In this particular situation, the authors propose improving atrioventricular synchrony by pharmacological prolongation of the PR interval, or, when unsuccessful, by ablation of the atrioventricular junction. This attitude was adopted in 5 cases when pacing with an AVD programmed between 20 and 70 ms, provided only a partial haemodynamic result. Improved atrioventricular synchrony allowed selection of an AVD of 100 to 150 ms, so reducing the gradient by more than 70%, compared with 30% under the initial programming parameters. The authors conclude that a positive and sustained benefit of cardiac pacing can only be obtained by an individualised regulation of pacing parameters to optimize atrioventricular synchrony which seems to be the key element of success. PMID- 7487304 TI - [Surgical treatment of hypertrophic cardiomyopathy; techniques, indications and results]. AB - Many techniques have been proposed for the treatment of hypertrophic cardiomyopathy over the last 35 years: myotomy, septal myotomy-myectomy, isolated mitral valve replacement with a low profile prosthesis, cardiac transplantation. Usually, the patients referred for surgery are those who do not respond or are resistant to medical therapy (patients in NYHA classes III or IV). The usual indication for myomectomy is a significant subaortic gradient and major septal hypertrophy (> 18 mm). When mitral regurgitation is severe or organic, mitral valve replacement is associated. When septal hypertrophy is moderate (< 18 mm) or not evenly distributed, or after ineffective myomectomy, mitral valve replacement may be proposed. Cardiac transplantation is only considered when all medical and surgical possibilities have been exhausted. Analysis of the results of surgery (over 1,000 published cases) is hindered by the variability of the techniques employed, the indications and experience of the different groups. The operative mortality has significantly decreased (25% in the 1960s to 2 to 11% at present). The complications of myomectomy are mainly postoperative ventricular septal defects and atrioventricular block, some of which require implantation of a pacemaker. Peroperative transoesophageal echocardiography could help to reduce the operative risk even further. Surgery improves symptoms and increases exercise capacity. The benefits seem greater, more frequent and longer lasting than with medical therapy. Surgery should not, however, be considered to be curative as some patients remain at risk of developing symptoms related to physiopathological phenomena other than intraventricular obstruction (poor left ventricular filling, myocardial ischaemia, arrhythmias). Although some workers suggest improved survival, there have been no controlled trials on this subject.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7487305 TI - [Cardiac pacing and cardiac insufficiency]. AB - Permanent Dual-chamber cardiac pacing has been proposed as treatment of advanced cardiac failure due to ischaemic or primary dilated, hypokinetic cardiomyopathies, since 1986. After encouraging initial results in 1990, the most recent studies have reported less convincing results, justifying the need for prospective, randomised studies. The physiopathological mechanisms underlying the haemodynamic improvement remain unknown. PMID- 7487306 TI - [Genetics of dilated cardiomyopathies]. AB - So-called idiopathic dilated cardiomyopathy is a multifactorial condition. Its familial nature is increasingly commonly observed in genealogical studies. The Cardiomyopathy Working Group of the French Cardiological Society has recensed 31 families with two or more members with dilated cardiomyopathy. This led to the search for genetic abnormalities in this condition, as it has been performed in other diseases, especially primary hypertrophic cardiomyopathies. PMID- 7487307 TI - [Antiarrhythmia agents in heart failure]. AB - Supraventricular and ventricular arrhythmias are extremely common in cardiac failure. No antiarrhythmic agent has been shown to improve survival in atrial fibrillation with left ventricular systolic dysfunction. In symptomatic ventricular tachycardia or fibrillation, treatment is generally determined by the ejection fraction (low EF: amiodarone; intermediate EF: amiodarone and/or betablocker; normal EF: most antiarrhythmics active at ventricular level). In asymptomatic ventricular arrhythmias, the prescription of antiarrhythmic drugs should be restrictive as few studies have demonstrated any benefits and the pro arrhythmic effects are dangerous. No medication has been shown to be of real value in ventricular tachycardia complicating hypertrophic cardiomyopathy. PMID- 7487308 TI - [Classic treatment of chronic heart insufficiency. What if new?]. AB - The aims of treatment of chronic heart failure are to improve the symptoms and the quality of life, reduce mortality and prevent left ventricular dysfunction. Before the first symptom occurs, neurohormonal activation takes place (increased catecholamines and atrial natriuretic peptide levels). Diuretics improve symptoms and are irreplaceable for the elimination of salt and water overload. Loop diuretics are used more often than the thiazides. Their deleterious effects on electrolyte balance are well known. The fact that they activate the renin angiotensin system is a more recent acquisition; the increase in plasma renin activity is a poor prognostic factor. Diuretics potentialize the vasodilator effect of angiotensin converting enzyme inhibitors which inhibit the neurohumoral activation induced by the diuretics. This therapeutic association is very logical, effective and allows reduction in the dosage of the diuretic. To date, there are no large scale controlled studies of the effects of diuretics on mortality. Spironolactone corrects hypokalaemia and hypomagnesaemia induced by loop diuretics. Moreover, it has been shown experimentally in renovascular hypertension and in hyperaldosteronism, that this molecule can prevent myocardial fibrosis, a factor which leads to ventricular dysfunction. The RALES study will analyse the effect of associating spironolactone to diuretic and ACE inhibitor therapy on the mortality of patients in NYHA classes III-IV. The value of digitalis in heart failure patients with sinus rhythm is a classical controversy. Digitalis has a positive inotropic effect (inhibition of NaK-dependent ATPase). More recently, a favourable neurohormonal effect has been reported; digitalis decreases the activation of the sympathetic and renin-angiotensin systems.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7487309 TI - [Treatment of chronic heart failure: current views]. AB - New agents for treating chronic heart failure include angiotensin converting enzyme (ACE) inhibitors, betablockers and phosphodiesterase inhibitors. The ACE inhibitors represent the major therapeutic advance of the 1980-1990 decade. This is the most effective class of drugs on survival, whatever the stage of heart failure and it shows the evolution towards symptoms in asymptomatic patients. Studies currently under way are evaluating the dose-effect relationship of ACE inhibitors. Betablockers improve the quality of life and physical performance but a benefit on mortality has not been shown in two recent trials. Phosphodiesterase inhibitors improve quality of life and physical performance at the price of an increase in mortality. Therefore, they are not indicated in the treatment of heart failure. However, new molecules such as vesnarininone or pimobendan are under trial. Finally, in the next few years, the introduction of antagonists to Angiotensin II receptors is eagerly awaited. PMID- 7487310 TI - [Myocardial revascularization and ischemic heart failure]. AB - Ischaemic heart failure is the cause of an increased number of hospital admissions and death rate. In severe heart failure and cardiogenic shock, coronary reperfusion associated or not with aortic balloon pump counter pulsation, seems to be beneficial. Moreover, it has recently shown that coronary bypass surgery may be a reasonable alternative to cardiac transplantation in coronary patients with severe heart failure. In the chronic phase, two physiopathological conditions have been proposed to explain the mechanism of ischaemic left-ventricular dysfunction: myocardial hibernation and stunning. Only hibernation requires revascularisation to avoid irreversible myocardial damage. The CASS group and others have demonstrated a real benefit from surgery although the operative risk is greater. Angioplasty has also been shown to be effective in terms of primary success with a relatively low complication rate. Compared one with another, angioplasty seems to provide better immediate results whereas surgery is associated with more long-term benefits. Myocardial revascularisation is therefore part of the therapeutic arsenal in cases of ischaemic heart failure. PMID- 7487311 TI - [Adrenergic betablockers and heart failure]. AB - Several studies have shown that beta-blockade could provide functional benefit in heart failure, suggesting a deleterious role for the compensatory sympathetic stimulation. Betablockers induced benefit could result from either antagonism of myocardial beta-adrenergic stimulation or, on the contrary a paradoxical increased cardiac beta-adrenergic responsiveness secondary to beta-adrenergic receptor up-regulation. Two recently completed large scale multicentric placebo controlled studies, the MDC trial with metoprolol and CIBIS with bisoprolol, have confirmed that beta-blockade could functionally improve patients with heart failure. The observed survival improvement in non-ischaemic patients was observed only in the CIBIS trial. This result requires confirmation by additional studies. PMID- 7487312 TI - [Anticoagulant treatment and dilated cardiomyopathy]. AB - The prevalence of intracardiac thrombi in patients with dilated cardiomyopathy is very variable from one study to another, but is generally high: 20 to 25% for interatrial thrombi and 50% for intraventricular thrombi. There is also a high incidence of left atrial spontaneous contrast (30-40%). Left atrial thrombosis or spontaneous contrast is more common in atrial fibrillation, when the LA diameter is increased with low velocity intra-left atrial blood flow on Doppler examination and when there are disturbances of haemorrheological factors (increased fibrinogen levels and plasma viscosity). Ventricular thrombi are more common when the fractional shortening is decreased (< 11% = 80% of thrombi). The frequency of embolism is controversial. It varies between 1.4 and 12 events per 100 patients per year. The risk of complications is higher in patients with intracavitary thrombosis. The number of cerebral haemorrhagic complications in patients on long-term oral anticoagulants is far from negligible. Large prospective multicenter trials should be instituted, as for atrial fibrillation to evaluate systematic anticoagulation with respect to the risks. In the meantime, it would seem to be prudent to prescribe oral anticoagulants to all patients with dilated cardiomyopathy with an intracardiac thrombus and/or atrial fibrillation, and to perform echocardiography regularly when left ventricular function is very poor. PMID- 7487313 TI - [Calcium antagonists and treatment of chronic heart failure]. AB - The history of the treatment of heart failure may be divided into three stages, the consequences of different conceptions of the physiopathology of the disease, with diuretics to counteract salt and water retention, vasodilators to improve conditions of cardiac load, angiotensin converting enzyme inhibitors to limit the effect of neurotumoral and sympathetic activation. There are two main reasons for using calcium antagonist in heart failure, the first being arterial vasodilatation leading to improve systolic function and the second being the beneficial effect on ventricular relaxation. However, their use has been controversial because of the results obtained with the first generation of these drugs. New molecules derived from dihydropyridine have been developed. Clinical trials with these second generation calcium antagonists are analysed. PMID- 7487314 TI - [The timing of cardiac transplantation]. AB - The considerable progress realised over the last 20 years in the domain of cardiac transplantation has had, as a corollary, an ever increasing demand and a cruel shortage of available grafts, responsible for a high mortality of some of the candidates on the waiting list. This situation justifies a review of the objective criteria of eligibility for a more pertinent attribution of donor organs. A review of the recent literature suggests a logical process in the evaluation of candidates. The first step consists of optimisation of medical therapy. This allows classification of patients with respect to the clinical condition obtained, which may be "critical", "unstable" or "stable" and thereby retain the indication for transplantation in the first two groups in the absence of a contra-indication. The timing of the transplantation is more difficult to determine for the 60% or so of patients with a low ejection fraction and who have been stabilised. The measurement of peak VO2 on exercise, which appears to be the most powerful prognostic variable in these patients, with respect to a normal subject of the same age, allows identification of urgent indications for transplantation. Moreover, a 3 monthly follow-up of peak VO2 of patients on the waiting list or deferred also enables reconsideration of their inscription or non inscription. PMID- 7487315 TI - [Heart transplantation, cardiomyoplasty or artificial heart?]. AB - This is a difficult question and the answer is uncertain. The authors review the state of the art of the three methods in 1993. Cardiac transplantation seems to have attained its maturity. The annual number of transplant operations is stagnant and the results progress little. Functional rehabilitation is excellent, the essential immunosuppression which has not changed in principle over the last 12 years, remains prejudicial. Cardiomyoplasty is an attractive concept with difficult surgical indications (Stage III, moderately dilated cardiomyopathy with good right ventricular function without arrhythmias, pulmonary hypertension or mitral regurgitation), a delayed efficacy, a hospital mortality comparable with that of transplantation and a similar survival rate. The objective results are not as good as the more subjective functional improvement. This limited experience (about 500 patients in 50 centers throughout the world, 70% of whom are European) should be continued and evaluated in the centers which initiated it. The artificial heart is only a temporary though essential therapeutic option in certain extremely urgent situations. It is a form of circulatory assistance, ranging from the simple univentricular accessory pump to the univentricular (Novacor) or biventricular (Jarvik) heart, in a rapidly evolving technology with problems of energy sources, marketing, cost and also clinical management which is often difficult especially with respect to coagulation. What do the next ten years hold in store for us? A nex immunosuppressor or the xenograft? A more efficient cardiomyoplasty with more precise medications? A totally implantable autonomous artificial heart? Can economic considerations accompany this development? This is undoubtedly the deepest source of concern for the future. PMID- 7487316 TI - [Value of coronary angiography in the diagnosis of coronary artery disease of the transplanted heart. Coronary angiography and arteriosclerosis of the graft]. AB - The diagnostic value of coronary angiography, a widespread method of detection of transplant coronary artery disease, was studied in 17 cardiac transplant patients with reference to histological examination. In the 6 coronary segments studied, the only significant but weak correlation that was found was for the distal left anterior descending artery: the correlations were not statistically significant in the other 5 segments. Coronary angiography underestimated lesions and false negative results were frequently reported (66 and 27% respectively). The limitations of coronary angiography may be explained by the technical artefacts related to both methods of evaluation and the anatomically diffuse and distal nature of transplant coronary artery atherosclerosis. A more reliable diagnostic method would seem to be required in view of the clinical importance of this pathology. PMID- 7487317 TI - [Sustained ventricular tachycardia: an evaluation of long-term prognosis]. AB - The authors analysed the clinical and paraclinical variables of 116 patients admitted to the Hopital Cardiologique de Lyon between 1986 and 1990 with sustained ventricular tachycardia without cardiocirculatory arrest in order to determine the long-term outcome and the prognostic factors of death and recurrence. The average age of the patients was 56 +/- 15 years (mean +/- SD) and 83% were men. The mean ejection fraction was 39 +/- 15%. Sixty-five had previous myocardial infarction (group I); 30 (group II) had dilated cardiomyopathy (n = 21), right ventricular dysplasia (n = 4), hypertrophic cardiomyopathy (n = 2), congenital (n = 2) or valvular (n = 1) heart disease. Group III comprised 21 patients with no apparent cardiac disease or isolated mitral valve prolapse. Brief syncope was reported in 12 cases. The paraclinical investigations showed 46 patients (66%) with at least two criteria of positivity for ventricular late potentials; Holter recording showed doublets or runs of VES in 46% of cases and sustained or non-sustained VT was induced during exercise testing in 16 patients (22%). Programmed ventricular stimulation triggered VT in 85%, 79% and 61% of patients in groups I, II and III respectively. The patients were treated with amiodarone in 65 cases, a betablocker in 25 cases, catheter ablation of the origin of the tachycardia in 12 cases, antiarrhythmic surgery in 6 cases, coronary bypass grafting in 5 cases (with an associated antiarrhythmic procedure in 3 cases). An automatic defibrillator was implanted in 9 patients. The average follow-up period was 32 months (range: 17 days to 65 months).(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7487318 TI - [Significance of lymphoplasmocytic infiltration in arrhythmogenic right ventricular dysplasia. Apropos of 3 own cases and review of the literature]. AB - Three patients with typical histological signs of arrhythmogenic right ventricular dysplasia progressed to fulminating heart failure in 1 case and unexplained sudden death in the other two. There were signs of superacute myocarditis in the first case, previous healed pericarditis in the second and chronic myocarditis in the third case. These cases suggested the presence of an inflammatory process complicating the substrate of the dysplasia. Arrhythmogenic right ventricular dysplasia seems to be a developmental defect which is complicated in 50 to 70% of patients, according to a review of 74 cases reported in the literature, by a varying degree of myocarditis suggesting particular susceptibility of these patients to infection and explaining the presence of unusual amounts of fibrous tissue in some and the so called "progressive" nature of the disease in other patients. PMID- 7487319 TI - [Quantification and monitoring of vascular resistance in the lower limbs by the Doppler method (animal model)]. AB - The object of this study was to define and validate a non-invasive method of evaluation and monitoring of vascular resistances in the leg. Blood flow velocity was measured by Doppler ultrasound in an animal model (ewe) with similar blood flow characteristics in the lower limb as man and allowing access to the required invasive measurements for validation of the method (pressure and flow). Vascular resistances distal to the measuring point (femoral, for example) were assessed using the resistance index R = D/S, S being the peak systolic deflection and D that of diastolic reflux of the Doppler spectral analysis of flow in the femoral artery. The values and variations of this resistance index were compared with the vascular resistances calculated from measurements of pressure and flow at the point of Doppler sampling and expressed in mmHg/ml/min. Femoral flow was measured by Doppler ultrasound (Doppler-echo), and mean pressure by an arterial catheter introduced into the abdominal aorta. Compression of the lower limb veins induced a venous return resulting in a reduction of cardiac output and femoral flow. During compression, femoral flow decreased by an average of 29% (p < 0.001) although mean pressure and heart rate did not change significantly. The femoral resistance index (Rf) increased by an average of 37.5% (p < 0.01) and vascular resistances increased by 45.9% (p < 0.01). Injection of 1 mg adrenaline induced peripheral vasoconstriction with an increase in blood pressure and a decrease in heart rate and femoral flow.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7487320 TI - [Nosology of atrial fibrillation. Attempt at clarification]. AB - Atrial fibrillation is the subject of much clinical interest as it is the arrhythmia responsible for the greatest number of hospital admissions. The arrhythmia working group of the French Society of Cardiology held a symposium on atrial fibrillation in order to establish certain definitions and to assess the value of a classification of atrial fibrillation. This was also the occasion to review a number of known facts and points which remain obscure, concerning this common arrhythmia. A therapeutic strategy is proposed. PMID- 7487321 TI - [Calcified proliferation of the descending thoracic aorta]. AB - Endoaortic calcific proliferation is a rare condition which causes progressive occlusion of the aortic lumen by endoluminal concretions. Symptoms are related to the resulting ischaemia and systemic hypertension caused by increased cardiac afterload. The authors report an exceptional case with a descending thoracic aortic lesion causing symptoms suggesting medullary ischaemia by partial occlusion of arteries supplying the spinal cord. The physiopathological mechanisms and therapeutic approaches are discussed with respect to a review of the literature. PMID- 7487322 TI - [Left ventricular thrombosis complicating systemic lupus erythematosus]. AB - The authors report an isolated pediculated thrombus in the left ventricle of a young 14 year old girl with systemic lupus erythematosus with antiphospholipid antibodies without any other cardiovascular abnormality, especially ventricular wall motion abnormalities. After surgical ablation of the thrombus, the patient was followed up to avoid recurrence. This type of cardiac lesion (ventricular thrombosis without underlying myocardial disease) is exceptionally rare. Echocardiographic follow-up after surgical ablation showed no recurrence of thrombosis after four years. PMID- 7487323 TI - [Pierre-Yves Hatt (1921-1994)]. PMID- 7487324 TI - [Prescription of hormonal replacement therapy: a new way to prevent heart diseases]. PMID- 7487325 TI - [Changes in blood cholesterol levels over a 10-year period; value of a single sample for predicting future blood cholesterol levels]. AB - The aim of this study was to report the serum cholesterol changes over a 10 year period in 12,238 subjects aged 4 to 64 years (mean 28 +/- 14 years) based on 3 health check-ups at an average of 5.5 yearly intervals between 1973 and 1989, and to determine the value of a single sample for predicting the serum cholesterol level at 5 and 10 years, and the influence of blood pressure and Quetelet index on this predictability. After identification of the influencing factors, the different variables were adjusted using a step-by-step regression analysis. The correlation coefficients calculated between the adjusted cholesterol level at the first examination and that measured at 5 and 10 years, were all significant (0.38 to 0.59) and varied with age at the time of the first examination and gender. The positive predictive value of having a cholesterol level higher than the 90th centile at 5 and 10 years when it was already higher than this value at the first examination varied from 26 to 46% respectively with respect to the subgroups. The sensitivity of the test was 25 to 48%. The negative predictive value and specificity were 93 to 95%. The lowering of this threshold to the 80th centile increased the positive predictive value from 35 to 45% and decreased the specificity from 94 to 87% for the whole population. When the first two sampling results, five years apart, were taken into consideration simultaneously, the predictive value of having a raised cholesterol level at 10 years increased from 35 to 61%.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7487326 TI - [4-year mortality of heart failure with intact systolic function: a retrospective studies]. AB - Heart failure with normal systolic function has been recognised in 30-40% of patients investigated for congestive heart failure. The authors undertook a retrospective study of global and cardiovascular mortality at 4 years in two groups of patients with a history of congestive heart failure documented by equilibrium angioscintigraphy: group I (n = 109) with poor left ventricular systolic function (EF = 26 +/- 9%) and group II (n = 40) with normal systolic function (EF = 58 +/- 8%). The 4 year cardiovascular mortality was less in group II (21.6% vs 49%; p < 0.004) whereas the global mortality was comparable in the two groups. The only discriminating parameters for cardiovascular mortality between the two groups were age of more than 60 years and values of diastolic blood pressure exceeding 90 mmHg which were associated with a higher mortality in the group with normal systolic function. These results indicate a better prognosis in terms of cardiovascular mortality from congestive heart failure in patients with normal systolic function compared with those with poor systolic function. PMID- 7487327 TI - [Influence of the valve position on the complications related to pericardial Carpentier-Edwards valve after 8 years]. AB - Between July 1984 and December 1991, 541 Carpentier-Edwards pericardial prostheses were implanted in 536 patients (420 aortic and 121 mitral prostheses). The operative mortality was 2.9% for aortic and 3.3% for mitral valve replacement. All but 8 patients were followed up for an average of 3.9 years, making a total follow-up of 1930 patient-years. There were 61 deaths during follow-up (41 AVR, 20 MVR). The actuarial 8 year survival was 80 +/- 6% for AVR and 74 +/- 10% for MVR. There were 15 deaths related to the prostheses. The probability of absence of valve-related mortality was 97 +/- 3% for AVR and 92 +/ 6% for MVR (NS). The valve-related complications included 15 thromboembolic events (AVR: 12; MVR: 3), 10 endocarditis (AVR: 7; MVR: 3), 8 anticoagulant related haemorrhages (AVR: 4; MVR: 4), 4 degeneration (AVR: 2; MVR: 2). There were no cases of cusp tear. The probability of absence of thromboembolic events at 8 years was 96 +/- 3% for AVR and 96 +/- 4% for MVR (NS), of absence of endocarditis 97 +/- 2% for AVR and 97 +/- 3% for MVR (NS) and of absence of valve degeneration 99 +/- 1% for AVR and 99 +/- 2% for MVR (NS). These results show that the position of the bioprosthesis had no influence on valve-related morbidity and that pericardium is a satisfactory tissue for the manufacture of mitral bioprostheses. PMID- 7487328 TI - [Outcome of arrhythmogenic right ventricular dysplasia. Apropos of 4 cases]. AB - The authors propose a classification of the outcome of arrhythmogenic right ventricular dysplasia with reference to 4 selected cases with a follow-up period of over 9 years. In type I, the left ventricular ejection fraction is normal (EF > 50%) and the risk, exclusively arrhythmic, can be controlled by appropriate antiarrhythmic therapy. This is the commonest form of arrhythmogenic right ventricular dysplasia with different varieties according to the degree of dilatation of the right ventricle. In type II, there is a variable degree of left ventricular involvement (30 < EF < 50%) either by extension of a comparable disease process as observed in the right ventricle or by an isolated or superimposed phenomenon of myocarditis. This form is stable and may remain stable for many years providing the arrhythmias are correctly treated. In type III, progressive degradation of the myocardium is observed over a period of about 10 years with a clinical presentation comparable to that of certain arrhythmogenic dilated cardiomyopathies which are often hereditary. In this case, the patients have an arrhythmic risk associated with that of cardiac failure which becomes progressively irreversible. The histology shows interstitial fibrosis with biventricular lymphocytic infiltration suggesting an autoimmune phenomenon. Therefore, the classification of cases of arrhythmogenic right ventricular dysplasia depends on the potential evolutivity of the lesions. When the patient is seen in the early stages of the disease, the prognosis should be garded, especially in a hereditary form. PMID- 7487329 TI - [Long-term results of cardiac transplantation]. AB - Between 1982 and 1992, 73 patients who had undergone cardiac transplantation and survived the hospital period, were followed up. The actuarial survival was 86%, 65% and 40% at 1, 5 and 7 years. The main causes of the 14 secondary deaths were infection (4), acute rejection (3) and cancer (3). Survival was complicated by acute rejection (1.07 episodes/patient), infection (0.7 episode/patient), cancer, hypertension and renal failure, graft dysfunction and other more secondary side effects. After analysing all the complications, the authors evaluated the quality of long-term survival after cardiac transplantation which allowed one patient out of two to return to normal living but with the threat of secondary graft dysfunction. PMID- 7487330 TI - [Balloon angioplasty for restenosis of coronary endoprostheses]. AB - Between March 1991 and June 1994, the authors treated restenosis (> or = 50%) of coronary endoprostheses in 17 men aged 39 to 72 years by balloon angioplasty. The stents had been implanted for de novo stenosis in 14 patients, restenosis in 1 patient and for acute occlusion in 2 patients. The vessels treated were the left anterior descending artery (1 case), the left circumflex artery (2 cases), the right coronary artery (7 cases) and venous bypass grafts (7 cases). The endoprostheses used were 5 Wallstents in 4 patients, 13 Wiktor stents in 11 patients and 2 Palmaz-Schatz in 2 patients. Clinically, the patients with restenosis presented with class II angina (6 patients), class III (8 patients) and class IV (3 patients) of the Canadian Cardiovascular Society Classification. Conventional balloon angioplasty was performed 17 months (range: 3 to 87 months) after stent implantation and resulted in immediate clinical and angiographic success (residual stenosis < or = 50% on quantitative analysis) in all patients. In particular, there were no dissections. The minimal lumenal diameter (MLD) and percentage stenosis changed from 1.04 mm (range: 0.87 to 1.20) and 72% (range: 68 to 76) respectively before stent implantation to 2.83 mm (range: 2.46 to 3.20) and 23% (range: 20 to 27) after stent implantation. Restenosis resulted in a MLD and percentage stenosis of 0.89 mm (range: 0.77 to 1.01) and 73% (range: 69 to 78) respectively. After angioplasty of the stent, they returned to 2.68 mm (range: 2.29 to 3.06) and 27% (range: 23 to 30) respectively. The patients were not anticoagulated after angioplasty.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7487331 TI - [Comparative outcome of aortic valve endocarditis with or without annular abscess]. AB - Annular abscess is a not uncommon but serious complication of aortic valve endocarditis. The aim of this retrospective study was to evaluate the prognosis of aortic valve endocarditis with and without annular abscess. Between January 1981 and 1989, 122 consecutive cases of aortic endocarditis fulfilling the diagnostic criteria of Duke University were admitted to hospital. Group I included 40 cases with aortic ring abscess confirmed at surgery, in 35 patients; group II comprised 43 cases of operated aortic valve endocarditis without annular abscess in 41 patients and group III comprised 38 cases of aortic valve endocarditis treated medically without echocardiographic or angiographic signs of annular abscess in 36 patients. The patients in group III were significantly older than those in group I (57 +/- 14 years vs 44 +/- 17 years; p < 0.001). From the clinical point of view, endocarditis of prosthetic valves was slightly more common, but without reaching statistical significance, in group I, but the abscess was associated with more severe cardiac failure. Systemic embolism, atrioventricular block and pericardial effusion were equally common in the three groups. On the other hand, endocarditis with annular abscess was more often the result of infection with streptococci A, B, C or pneumoniae, than forms without abscess (22.5% vs 5% and 3% respectively in the 3 groups; p < 0.05). Of the patients treated surgically, destructive lesions of the valves were more common in cases of abscess (57.5% vs 35%; p < 0.05): the hospital mortality was higher in cases of abscess (17.5% vs 7%).(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7487332 TI - [Characteristics and influence of different clinical forms on the development and prognosis of carotid sinus syndrome. Apropos of 215 cases]. AB - The carotid sinus syndrome is a well-known cause of syncope: the cardio inhibitory forms are the easiest to diagnose and probably the easiest to treat. However, the vasodepressive forms are as common but their outcome is mainly unknown. Eight hundred and fifty-three patients underwent endocavitary electrophysiological studies with invasive blood pressure measurement for unexplained syncope between October 1984 and January 1990. A carotid sinus syndrome was diagnosed in 215 cases. Fifty-two patients (24.2%) had a cardio inhibitory form (ventricular standstill > or = 3 s during carotid sinus massage), 40 (18.6%) had a pure vasodepressive form (isolated fall of systolic blood pressure > 50 mmHg during massage) and 123 patients (57.2%) had a mixed form. The average age was 74.1 +/- 9.7 years with no difference between the different forms. A number of parameters was different on the cardio-inhibitory and vasodepressive forms: the number of men (75.6 vas 24.4%; p < 0.05) and the number of syncopes (83.3 vs 60%; p < 0.02) were greater in the cardio-inhibitory form; the vasodepressive forms were more often associated with a history of transient ischaemic attacks (15 vs 0%), a poor cardiovascular status (hypertension: 47.5 vs 15.7%; p < 0.01), coronary artery disease (47.5 vs 25.5%; p < 0.05), cardiac failure (27.5 vs 11.7%; p < 0.05), induction of sustained supraventricular tachycardia (50 vs 23.5%; p < 0.05) and a greater pacemaker effect (53.6 vs 34.8 mmHg; p < 0.01); of the 191 patients (84.9% of the population) followed up for an average of 21.2 months, 168 received treatment: implantation of a cardiac pacemaker in 108 patients, reduction of antihypertensive and/or potentially bradycardia-inducing drugs in 30 patients, prescription of antiarrhythmic therapy, in 30 patients.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7487333 TI - From growth to autolysis: the murein hydrolases in Escherichia coli. AB - Murein hydrolases cleave bonds in the bacterial exoskeleton, the murein (peptidoglycan) sacculus, a covalently closed bag-shaped polymer made of glycan strands that are crosslinked by peptides. During growth and division of a bacterial cell, these enzymes are involved in the controlled metabolism of the murein sacculus. Murein hydrolases are believed to function as pacemaker enzymes for the enlargement of the murein sacculus since opening of bonds in the murein net is needed to allow the insertion of new subunits into the sacculus. Furthermore, they are responsible for splitting the septum during cell division. The murein turnover products that are released during growth are further degraded by these (1 --> 6)-anhydromuramic acid derivatives by an intramolecular transglycosylation reaction. PMID- 7487334 TI - Lipopolysaccharide of Rhodospirillum salinarum 40: structural studies on the core and lipid A region. AB - The structural elucidation of lipid A of the cell wall lipopolysaccharide (LPS) of Rhodospirillum salinarum 40 by chemical methods and laser desorption mass spectrometry revealed the presence of a mixed lipid A composed of three different 1,4'bisphosphorylated beta (1 --> 6)-linked backbone hexosaminyl-hexosamine disaccharides, i.e. those composed of GlcN --> GlcN, 2,3-diamino-2,3-dideoxy-D Glc-(DAG --> DAG, and DAG --> GlcN. Lipid A of R. salinarum contained preferentially 3-OH-18:0 and 3-OH-14:0 as amide-linked and cis delta 11-18:1 and c19:0 as ester-linked fatty acids. The mass spectra of the liberated acyl-oxyacyl residues proved the concomitant presence of 3-O-(cis delta 11-18:1)-18:0 and 3-O (c19:0)-14:0 as the predominating diesters in this mixed lipid A. The glycosidically linked and the ester-linked phosphate groups of the backbone disaccharide were neither substituted by ethanolamine, phosphorylethanolamine, nor by 4-amino-4-deoxy-L-arabinose, in contrast to most of the enterobacterial lipid As. In the core oligosaccharide fraction, a HexA (1 --> 4)HexA(1 --> 5)Kdo trisaccharide was identified by methylation analysis. The terminal HexA (hexuronic acid) is possibly 4-OMe-GalA, a component described here as an LPS constituent for the first time. LPS of R. salinarum showed a lethality in C57BL/10 ScSN (LPS-responder)-mice) of an order of 10(-1)-10(-2) of that reported for Salmonella abortus equi LPS, and it was also capable of inducing TNF alpha and IL6 in macrophages of C57BL/10ScSN mice. PMID- 7487335 TI - Cyclodextrins for growth of Helicobacter pylori and production of vacuolating cytotoxin. AB - Growth of Helicobacter pylori in liquid culture requires the addition of media supplements that often interfere with subsequent purification of bacterial antigens. In order to determine whether cyclodextrins can substitute for conventional H. pylori growth supplements, we cultured H. pylori in the presence of five commercially available cyclodextrins. The effect of these compounds on the production of the vacuolating cytotoxin antigen was evaluated. Several cyclodextrins supported flourishing growth and permitted the consistent production of vacuolating cytotoxin. These data suggest that Brucella broth supplemented with cyclodextrins is an improved medium for bacterial culture and industrial production of H. pylori antigens. PMID- 7487337 TI - Genetic epidemiology in psychiatry. Taking both genes and environment seriously. PMID- 7487336 TI - An essential role for DsbA in cytochrome c synthesis and formate-dependent nitrite reduction by Escherichia coli K-12. AB - An Escherichia coli K-12 mutant, isolated on the basis of its inability to catalyze formate-dependent nitrite reduction, was characterized. The mutant was defective in the synthesis of all known c-type cytochromes during anaerobic growth. The mutation was localized by conjugation, transduction, and Southern blotting experiments to the dsbA gene at minute 87 on the E. coli chromosome and was complemented by the wild-type allele. Both DsbA and the recently described DipZ protein were shown to be essential for cytochrome c synthesis, suggesting that they act sequentially in a pathway for cytochrome c assembly in the E. coli periplasm. PMID- 7487338 TI - Antisocial behavior. PMID- 7487339 TI - Differential heritability of adult and juvenile antisocial traits. AB - BACKGROUND: Studies of adult antisocial behavior or criminality usually find genetic factors to be more important than the family environment, whereas studies of delinquency find the family environment to be more important. We compared DSM III-R antisocial personality disorder symptoms before vs after the age of 15 years within a sample of twins, rather than comparing across studies. METHODS: We administered the Diagnostic Interview Schedule Version III-revised by telephone to 3226 pairs of male twins from the Vietnam Era Twin Registry. Biometrical modeling was applied to each symptom of antisocial personality disorder and summary measures of juvenile and adult symptoms. RESULTS: Five juvenile symptoms were significantly heritable, and five were significantly influenced by the shared environment. Eight adult symptoms were significantly heritable, and one was significantly influenced by the shared environment. The shared environment explained about six times more variance in juvenile anti-social traits than in adult traits. Shared environmental influences on adult antisocial traits overlapped entirely with those on juvenile traits. Additive genetic factors explained about six times more variance in adult vs juvenile traits. The juvenile genetic determinants overlapped completely with genetic influences on adult traits. The unique environment (plus measurement error) explained the largest proportion of variance in both juvenile and adult antisocial traits. CONCLUSIONS: Characteristics of the shared or family environment that promote antisocial behavior during childhood and early adolescence also promote later antisocial behavior, but to a much lesser extent. Genetic causal factors are much more prominent for adult than for juvenile antisocial traits. PMID- 7487340 TI - Genetic-environmental interaction in the genesis of aggressivity and conduct disorders. AB - BACKGROUND: The purpose of this study was to determine the effect of an adverse adoptive home environment on adoptee conduct disorder, adult antisocial behavior, and two measures of aggressivity, all of which are behaviors that contribute to adult antisocial personality disorder and that also are associated with increased vulnerability to drug abuse and/or dependence. METHODS: The study used an adoption paradigm in which adopted offspring who were separated at birth from biologic parents with documented (by prison and hospital records) antisocial personality disorder and/or alcohol abuse or dependence were followed up as adults. They and their adoptive parents were interviewed in person. These adoptees were compared with controls whose biologic background was negative for documented psychopathologic behavior. Subjects were 95 male and 102 female adoptees and their adoptive parents. RESULTS: Multiple regression analysis was used to measure separately genetic and environmental effects. It showed that (1) a biologic background of antisocial personality disorder predicted increased adolescent aggressivity, conduct disorder, and adult antisocial behaviors, and (2) adverse adoptive home environment (defined as adoptive parents who had marital problems, were divorced, were separated, or had anxiety conditions, depression, substance abuse and/or dependence, or legal problems) independently predicted increased adult antisocial behaviors. Adverse adoptive home environment interacted with biologic background of antisocial personality disorder to result in significantly increased aggressivity and conduct disorder in adoptees in the presence of but not in the absence of a biologic background of antisocial personality disorder. CONCLUSIONS: Environmental effects and genetic environmental interaction account for significant variability in adoptee aggressivity, conduct disorder, and adult antisocial behavior and have important implications for the prevention and intervention of conduct disorder and associated conditions such as substance abuse and aggressivity. PMID- 7487341 TI - Genetic questions for environmental studies. Differential parenting and psychopathology in adolescence. AB - BACKGROUND: Recent genetic evidence suggests that the most important environmental influences on normal and pathologic development are those that are not shared by siblings in the same family. We sought to determine the relationship between differences in parenting styles and depressive symptoms and antisocial behavior in adolescence, and to compare the influence of these nonshared experiences with genetic influences. METHODS: We studied 708 families with at least two same-sexed adolescent siblings who were monozygotic twins (93 families), dizygotic twins (99 families), ordinary siblings (95 families), full siblings in step families (181 families), half siblings in step families (110 families), and genetically unrelated siblings in step families (130 families). Data on parenting style were collected by questionnaire and by video recording of interaction between parents and children. RESULTS: Almost 60% of variance in adolescent antisocial behavior and 37% of variance in depressive symptoms could be accounted for by conflictual and negative parental behavior directed specifically at the adolescent. In contrast, when a parent directed harsh, aggressive, explosive, and inconsistent parenting toward the sibling, we found less psychopathologic outcome in the adolescent. CONCLUSIONS: Parenting behavior directed specifically at each child in the family is a major correlate of symptoms in adolescents. Furthermore, harsh parental behavior directed at a sibling may have protective effects for adolescents, a phenomenon we call the "siblin barricade." PMID- 7487342 TI - Seasonal variation in plasma L-tryptophan availability in healthy volunteers. Relationships to violent suicide occurrence. AB - OBJECTIVE: To investigate the seasonal variation in levels of plasma L-tryptophan and competing amino acids (CAAs) in healthy humans in relation to climatic variables, total serum protein levels, and violent suicide occurrence. METHODS: Twenty-six healthy volunteers (13 men and 13 women; mean [+/- SD] age, 38.7 +/- 13.4 years) had monthly blood samplings for assays of L-tryptophan, valine, leucine, isoleucine, tyrosine, and phenylalanine during 1 calendar year. RESULTS: Significant annual rhythms were detected in L-tryptophan, the L-tryptophan/CAA ratio, phenylalanine, valine, and leucine, and semiannual rhythms in L-tryptophan values and in L-tryptophan/CAA ratios. Plasma L-tryptophan and the L tryptophan/CAA ratio were significantly lower in the spring than in the other seasons. The peak-trough differences in the yearly variation expressed as a percentage of the mean were 17.1% and 16.1% for L-tryptophan values and L tryptophan/CAA ratios, respectively. The amplitude of the yearly variation in all CAAs was low, ie, less than 7%. An important part of the variance in L-tryptophan availability (ie, 12% to 14%) could be explained by the composite effects of present and past climatic factors; higher ambient temperature and relative humidity in the face of lower air pressure are the most important predictors of low L-tryptophan availability. Important and positive time relationships were noted between total serum protein level and all amino acid concentrations, and a significant time relationship was also noted between the seasonal variation in L tryptophan availability and the occurrence of violent suicide in Belgium. CONCLUSION: Our results show a bimodal seasonal pattern in the availability of plasma L-tryptophan that matches seasonal patterns in the prevalence of violent suicide in the local population and depression in other studies. PMID- 7487343 TI - Evidence of chaotic mood variation in bipolar disorder. AB - BACKGROUND: Using long-term daily mood records obtained from patients with bipolar disorder and normal subjects, we sought to determine the temporal pattern of mood in bipolar disorder. METHODS: Time series of 1.0 to 2.5 years from seven rapid-cycling patients with bipolar disorder and 28 normal controls were obtained. These were evaluated with several techniques to identify whether the temporal pattern of mood originates from a periodic, a random, or a deterministic source. RESULTS: True cyclicity was not apparent in the power spectra of either the normal subjects or the patients with bipolar disorder. Instead, spectra with a broadband "l/f" shape were observed in both groups, and these spectra were significantly flatter in normal subjects (P = .02). Correlation dimension estimates are a measure of nonlinear deterministic structure, and convergent estimates could be obtained for six of the seven patients with bipolar disorder and none of the normal subjects (P < .001). Additional findings are consistent with these results. CONCLUSIONS: These studies indicate that mood in patients with bipolar disorder is not truly cyclic for extended periods. Nonetheless, self rated mood in bipolar disorder is significantly more organized than self-rated mood in normal subjects and can be characterized as a low-dimensional chaotic process. This characterization of the dynamics of bipolar disorder provides a unitary theoretical framework that can accommodate neurobiologic and psychosocial data and can reconcile existing models for the pathogenesis of the disorder. Furthermore, consideration of the dynamical structure of bipolar disorder may lead to new methods for predicting and controlling pathologic mood. PMID- 7487344 TI - Chaos and complexity. Can it help us to understand mood and behavior? PMID- 7487346 TI - Effect of outer membrane proteins (OMP) of Shigella on interleukin 2 (IL-2) production by spleen cells of mice. AB - The kinetics of interleukin (IL-2) release from mouse spleen cells incubated with different doses of outer membrane proteins (OMP) from Shigella was investigated. OMP induced very low activity of IL-2 after 2 and 4 h, and only a slightly higher level of the cytokine was detected after 6 h. However, IL-2 activity increased markedly after 20 and 24 h of incubation, and doses of 5 and 10 micrograms of OMP were found to be the most effective. Spleen cells cultured with OMP for 48 h contained reduced concentration of IL-2. PMID- 7487345 TI - Pindolol, 5-hydroxytryptamine, and antidepressant augmentation. PMID- 7487347 TI - Thymocyte-macrophage interactions. Interleukin 6 produced by activated thymocytes induces interleukin 1 production by macrophages. AB - Coculture of paraformaldehyde-fixed macrophages with syngeneic thymocytes resulted in synthesis of a lymphokine which triggered IL-1 production in untreated, peritoneal macrophages. The release of that lymphokine was MHC restricted, however, the stimulation of IL-1 production was not MHC-controlled event. Preliminary studies showed that the lymphokine had molecular weight (MW) of around 10-30 kDa, was heat stable (56 degrees C), resistant to reduction to DTT and exhibited a weak pyrogenic activity. Fusion of activated thymocytes with the thymoma BW5147 resulted in obtaining several hybridoma clones producing the lymphokine. Preparation of culture supernatants by means of Amicon membranes and Sephadex G-100 filtration allowed to determine the MW of the lymphokine as 25 kDa. Lastly, the lymphokine was identified as IL-6 by means of anti-IL-6 antibodies which blocked its function and by the use of IL-6-sensitive cell line 7TD1. PMID- 7487348 TI - Lactoferrin inhibits the effector phase of the delayed type hypersensitivity to sheep erythrocytes and inflammatory reactions to M. bovis (BCG). AB - Bovine lactoferrin (BLF) given into mice, sensitized to SRBC, together with the eliciting dose of antigen, inhibits very strongly the DTH reaction measured after 24 h by foot pad swelling. Administration of BLF at 48 or 24 h before eliciting the DTH reaction was not effective, however, BLF suppressed the reaction when given at the peak of the inflammatory process. The effects of BLF were strongest when the protein was injected intravenously. Intraperitoneal or intramuscular administrations of BLF were less inhibitory. In addition, BLF diminishes, although to a much lesser degree, the inflammatory reactions induced by BCG. The inhibitory action of BLF does not involve liver since treatment of mice with galactosamine does not reverse the inhibition. Studies on cytokine production revealed that peritoneal macrophages, derived from mice pretreated with LF, have an increased ability to produce in vitro IL-6 after induction with LPS. In addition, we demonstrated that inhibition of macrophage migration, mediated by migration inhibition factor, is abolished by BLF. Lastly, the inhibitory effect of BLF could not be transferred with serum from donors treated with BLF. In summary, the data reveal the inhibitory properties of LF, administered systematically, in relation to locally induced inflammation. PMID- 7487350 TI - Generation of anti-hapten T cell cytotoxicity in vivo. Relationship to contact sensitivity and the role of contrasuppression. AB - Immunization procedures that induce contact sensitivity to the trinitrophenyl (TNP) hapten in vivo were investigated for their ability to induce TNP-specific cytotoxic T lymphocytes in vivo. Spleen cells from C3H/HeN mice primed for CS responses either by the topical application of picryl chloride or by the adoptive transfer of PCL immune cells show little or no cytolytic activity in vitro against TNP-coupled target cells. Intravenous immunization with TNP-substituted syngeneic spleen cells, a procedure known to make animals unresponsive to agents normally inducing CS, also failed to induce cytolytic activity in spleen cells. However, both PCL sensitization and adoptive transfer, when combined with the injection of TNP-substituted syngeneic spleen cells, induce significant cytolytic activity against TNP-haptenated BW5147 target cells in vitro. Furthermore, i.v. injection of TNP-spleen cells with surface-bound immune complexes of the IgM or IgG1 isotypes, or with a monoclonal TNP-specific contrasuppressor T cell factor also induces strong antigen-specific cytolytic activity against TNP modified targets. TcsF bears serological determinants of T cell receptor alpha and beta chains and adheres to specific antigen columns. All these immunization regimens were shown to induce CS to TNP as well as the generation of contrasuppressor T cells. The CTL generated in the spleens of immunized mice are Thy1+ CD8+ T cells an are antigen-specific and genetically restricted. The implications of these results with respect to the mechanisms by which cytolytic responses are controlled in vivo is discussed. PMID- 7487349 TI - Interleukin 3-dependent, asialo-GM1-positive cell line with natural cytotoxic but without natural killer cytotoxic activity. AB - IL-3-dependent cell line (designated as A-3) from adherent spleen cells was developed. The surface phenotype of A-3 cells was analysed by flow cytometry and determined as ASGM1+, FcR+, Ia+. There was no expression of T cell markers (Th1 CD3-CD4-CD8-), B cell marker (surface Ig) as well as macrophage marker Mac-1 antigen. Although A-3 cells expressed ASGM1, they are lacking NK cytotoxic activity in 4 h Cr release assay with YAC-1 targets. In contrast, A-3 cells are endowed with NC cytotoxic activity as shown in 18 h Cr release assay with WEHI 164 target cells. PMID- 7487352 TI - Inhibitory activity of soluble material from sheep eosinophils for sheep complement. AB - Soluble material from frozen and thawed sheep eosinophils and sonicated extracts of eosinophils or their granules inhibited the alternative activation pathway dependent action of sheep complement for human erythrocytes sensitized by sheep antibody. Sephadex G-50 fractions of the sonicated eosinophil and granule extracts showed that inhibitory activity was associated with the void volume fraction, the fraction possessing the eosinophil peroxidase activity. PMID- 7487351 TI - Guinea pig antibody to C3 component of the sheep complement produced by immunization with a heat-inactivated sheep serum--alcian blue complex. AB - Normal sheep serum or serum heated at 56 degrees C for 30 min were added to alcian blue solutions. The optical density of alcian blue-heated sheep serum complexes formed were 2-4 times greater than those obtained with unheated serum. Most guinea pigs immunized with alcian blue-heated sheep serum complexes produced antibodies that precipitated the third component (C3) of sheep complement (C') from electrophoresed sheep serum. These antisera also hemagglutinated sheep erythrocyte--rabbit antibody complexes treated with sheep serum (C'). Guinea pigs immunized with alcian blue-normal sheep serum complexes did not react with electrophoresed sheep serum nor with sheep C' in the hemagglutination test. PMID- 7487353 TI - Immunosuppressive activity of C-terminal fragments of tachykinins. AB - The immunosuppressive activity of three pentapeptides related to the C-terminal fragments of tachykinins was investigated using the PFC (plaque forming cell) test. The peptides were of the general structure Phe-X-Gly-Leu-Met-NH2, with Tyr, Val and Ile in X position, respectively. The results were compared with those obtained previously for the substance P (SP) C-terminal pentapeptide. It was found that peptides containing aromatic residues in X position were more potent immunosuppressors than peptides containing aliphatic residues in the same position. The results also suggest that in mammalian organisms substance SP may be involved in immunomodulation which is rather not the case with other mammalian tachykinins (neurokinin A and neurokinin B). PMID- 7487354 TI - Immunosuppressive activity of analogs of tripeptide Lys-Arg-Pro with D-amino acid residues. AB - Synthesis of four analogs, containing D-amino acid residues, of the tripeptide immunosuppressor Lys-Arg-Pro is described. In addition, acetyl and amide derivatives of this peptide were synthesized. Biological effects of all peptides were studied using a delayed type hypersensitivity (DTH) test. It was found that incorporation of D-amino acid residues in definite positions of the peptide chain gives the substances with immunosuppressive activity. A strong immunosuppressive activity was observed also for all-D-analogs (D-Lys-D-Arg-D-Pro). PMID- 7487355 TI - Structural and immunochemical studies on O-specific polysaccharide of Proteus penneri strain 14. AB - The complete structure of the O-antigen of Proteus penneri strain 14, containing D-alanine and L-alanine was established using methylation, solvolysis with anhydrous hydrogen fluoride, partial acid hydrolysis, 1H- and 13C-NMR spectroscopy. The role of partial structures of the pentasaccharide repeating unit in manifesting serological specificity and cross-reactivity of this strain with some other bacteria is discussed. PMID- 7487356 TI - Influence of different temperature of cryopreservation on the proliferative potential of human bone marrow progenitor cells. Transplantological implications. AB - The influence of two different temperatures of cryopreservation of human bone marrow on the proliferative potential of granulocyto-monocytic (GM-CFU), erythroid (BFU-E) and megakaryocytic (CFU-Meg) progenitors was investigated. For this purpose, the human bone marrow cells were cryopreserved in the standard freezing medium supplemented with 10% DMSO and stored at -80 degrees C in a freezer or at -196 degrees C in a liquid nitrogen tank. Subsequently, cells were thawed, plated and stimulated in vitro to grow myeloid colonies. Unexpectedly, bone marrow cells stored at -80 degrees C formed about 60%, and those stored at 196 degrees C formed barely 8% of CFU-GM and BFU-E colonies respectively, in comparison to control non-frozen cells. CFU-Meg progenitors appeared to be the most sensitive to cryopreservation among all clonogeneic cells tested. The number of CFU-Meg derived colonies decreased to 20% in marrow cryopreserved at -80 degrees C and to about 2% in cells stored in liquid nitrogen at -196 degrees C. These data demonstrate the advantages of bone marrow storage at -80 degrees C over freezing in liquid nitrogen at -196 degrees C. Moreover, the increased sensitivity of CFU-Meg to cryopreservation could explain, at least partially, the clinical phenomenon of protracted thrombocytopenia observed in patients transplanted with the cryopreserved bone marrow cells. PMID- 7487357 TI - Absorption with K562 erythroblastoma cells as a means for discrimination between auto- and alloantibodies in sera of hypersensitized renal patients. AB - We applied K562 erythroblastoma cell line as a substitute for autologous lymphocytes in the absorption of autoantibodies from sera of 26 hypersensitized renal patients exhibiting high degree of cytotoxic reactivity with a panel of lymphocytes from unrelated individuals (panel reactivity, PRA). All of analysed sera contained allo-reactivity before absorption. After removal of autoantibodies, 85% of the sera still contained alloantibodies whereas in 15% of the sera no alloreactivity was detectable. Absorption of high PRA sera with K562 cells and with autologous lymphoblastoid cell lines derived by transformation with Epstein-Barr virus (EBV-LCL) gave substantially the same results. As autoantibodies have reportedly no effect on kidney graft survival, the autoabsorption of high PRA sera may increase the chance of patients in whom only autoantibodies were detected to receive the transplant. Application of K562 cells, instead of autologous lymphocytes/lymphoblasts, is recommended whenever high numbers of patient's cells are not available. It may be particularly suitable for routine clinical laboratories, not equipped and prepared for derivation and propagation of EBV-LCL. PMID- 7487360 TI - Influence of acute and chronic ethanol treatment on interferon production in mice. AB - The effect of acute (single dose) and long-term ethanol treatment of Swiss Albino and C3H/He mice on IFN production stimulated in vitro by NDV, PHA and ConA was examined. Liver organ cultures and spleen cells of mice which received intraperitoneally a single dose (2 g/kg of body weight) of ethanol (II group) or were drinking ethanol solutions instead of water for 4 weeks (III group) or for 8 weeks (IV group), and cells of control mice not treated with ethanol (I group) were cultivated in vitro. Some of the cultures were treated with 0.4% (w/v) ethanol or several IFN inducers NDV-R, PHA and ConA or incubated without any stimulators. The titers of IFN in supernatant fluids were measured. The results showed that cells from mice, which received a single ethanol injection, produced IFN spontaneously and this production was enhanced in the presence of 0.4% ethanol in culture medium. When spleen cell cultures of acute treated mice were induced with PHA and ConA, IFN titers lower than in cells of the control mice were produced. In contrast to these results, liver and spleen cells of mice after long term ethanol consumption produced significantly lower IFN titers than controls after NDV-R induction. Also spleen cells responded poorly both to PHA and ConA induction. These results demonstrate that long-term ethanol treatment can destroy IFN response to viral infection and can impair mitogen-induced IFN production. This suppression of IFN production by long term ethanol was similar in both strains of the mice examined. PMID- 7487358 TI - Determination of immunoglobulin class of alloantibodies in sera from hypersensitized renal patients. AB - 26 sera from hypersensitized renal patients were analysed for immunoglobulin class of alloantibodies. To assess the participation of IgM and IgG in alloreactivity, sera were treated with dithiothreitol (DTT) or Staphylococcus aureus protein A (SpA), respectively. Treatment with both DTT and SpA was also performed to test the possible contribution of other immunoglobulin classes. Immunological complexes which may mimic alloreactivity were excluded by polyethylene glycol (PEG) treatment. Immunochemically modified sera were tested against a panel of allogeneic B and T lymphocytes. IgG class alloantibodies reactive with B lymphocytes were found two times more frequently than those reactive with T lymphocytes. In 22% of sera only IgM alloantibodies were detected. No contribution of other immunoglobulin classes of immune complexes was seen. Our results show that DTT and SpA treatment are technically simple and reproducible methods of discriminating between IgG and IgM alloantibodies and can be recommended for routine use in clinical analysis of sera from hypersensitized renal patients awaiting kidney graft. PMID- 7487359 TI - Interferon production by peripheral blood cells of patients with alcoholic liver disease (ALD). AB - Blood samples from 29 healthy individuals (control), 16 chronic alcohol abusers without clinical symptoms of liver disease, 22 patients with alcoholic steatosis and 31 patients with alcoholic cirrhosis were obtained. The antiviral activity, only partially neutralized by polyclonal and monoclonal anti-HuIFN-alpha and monoclonal anti-HuIFN-gamma antibodies, was detected in sera of 38% healthy controls and of 64-77% alcoholic abusers without or with ALD. To measure the activity of the IFN system, leukocyte cultures of chronic alcohol abusers and control individuals were set up using a whole blood technique. Common IFN inducers: NDV, PHA and ConA were used. Blood leukocytes of patients with alcoholic steatosis and cirrhosis responded to NDV, PHA and ConA with significantly lower IFN levels than controls, whereas IFN response of alcoholics without liver diseases was lower only after stimulation with ConA. Regarding the lymphoproliferative test performed with PHA, a remarkably diminished lymphocyte response of patients with cirrhosis was observed as compared to control. Our data indicate that long term ethanol intake impairs the ability of leukocytes and their subpopulation to produce IFNs. The correlation between ALD progression and the defect in IFN production suggests that suppressive action of ethanol, liver injury and immune reactions involved in liver damage may be responsible for impairment of the IFN system in heavy drinkers. PMID- 7487361 TI - Involvement of heat shock proteins and cellular membranes in the development of thermotolerance. AB - A variety of cells subjected to hyperthermia acquire a transient increase in resistance (thermotolerance) to a subsequent heat treatment. The development of thermotolerance is nearly always accompanied by the presence of heat shock proteins (HSPs). However, other states of thermotolerance, some of which are independent of HSPs, have also been postulated. Recently, several studies have shown that cellular membranes, among other cellular structures, may be involved in the response of cells to heat and may constitute a target for the acquisition of thermal resistance. PMID- 7487362 TI - Specific pollen allergen activates eosinophils of the patient with chronic allergic contact urticaria. AB - The aim of the study was to evaluate the activation of eosinophils in an unique case of a young man with atopy manifested as chronic pollen contact urticaria. In order to reveal the role of eosinophils in that case, the study was performed by means of monoclonal antibodies EG2 and chemiluminescence. In addition, comparative electron microscopic study of peripheral blood and skin infiltrating eosinophils were performed for which the name ultrastructural morphometric analysis of intracytoplasmic eosinophil granules has been proposed. The results indicated, that 40% of peripheral blood eosinophils were activated spontaneously and they were more active than those in skin infiltrates. Specific pollen allergen caused activation of 100% of peripheral blood eosinophils. The study suggests presence of a systemic pattern of eosinophil activation in atopy. PMID- 7487363 TI - High serum levels of allergen specific IgG-4 (asIgG-4) for common food allergens in healthy blood donors. AB - High serum levels of asIgG-4 against common food allergens are found in many patients with symptoms suggesting food allergy. The same patients are frequently negative for allergen specific IgE (asIgE) against the same allergens. These data were frequently interpreted as suggestive of a role of asIgG-4 in food allergy. In order to evaluate this hypothesis we tested serum levels of asIgG-4 against food allergens in young blood donors without any signs or history of food allergy. Fifty young healthy male donors were evaluated. The serum levels of IgE, and asIgE and IgG-4 against 14 common food allergens were determined. The studies were carried out using commercially available 3M Diagnostics Systems kits. AsIgG 4 against food allergens were found in sera of 92% blood donors, and in 62% of these healthy persons the levels of asIgG-4 were higher than 10.0 micrograms/ml. In a small proportion of patients, high serum levels of IgE and asIgE against the same food and/or inhalant allergens were found. Common occurrence of asIgG-4 against food allergens in healthy persons (without any symptoms which could suggest allergy or food intolerance) argues against the possible participation of these antibodies in the pathogenesis of food allergy. It is possible that their occurrence is the result of immunization against food antigens (allergens). It remains to be resolved whether the presence of these antibodies represents an epiphenomenon or may have some other biological role. PMID- 7487364 TI - Immunological status of patients with denture stomatitis and yeast infection after treatment of maxillofacial tumors. AB - Therapeutic modalities employed in the treatment of maxillofacial malignancy, tumor resection, radiation and chemotherapy, can result in local and systemic changes in healthy oral tissues many years after anticancer therapy has been stopped. The immunologic status of a select group of nine individuals who underwent surgery of the maxillofacial region and after head and neck irradiation, with severe and persistent denture stomatitis was studied. A significant impairment of granulocyte and lymphocyte functions and increased concentrations of serum IgG, as compared to healthy controls, were found in the patients of the study group. It is concluded, that this group of patients show immunological abnormalities many years post therapy, which must be recognized by the clinician in order to effect a cure. PMID- 7487365 TI - Synergistic antiproliferative activity of tumor necrosis factor alpha (TNF-alpha) and lovastatin. AB - We assessed the antiproliferative effect of tumor necrosis factor alpha (TNF alpha) and lovastatin, an inhibitor of 3-hydroxy-3-methylglutaryl-coenzyme A (HMG CoA) reductase, alone and in combination, on two murine tumor cell lines. Recombinant TNF-alpha inhibited proliferation of murine MmB16 melanoma cells in a concentration-dependent fashion but stimulated growth of murine L1210 leukemia cells at 0.1 ng/ml concentration. Lovastatin inhibited proliferation both of murine MmB16 melanoma cells and of murine L1210 leukemia cells in a concentration dependent fashion. In combination with tumor necrosis factor alpha lovastatin inhibited synergistically growth of both cell lines as assessed by isobologram analysis. Our data show that lovastatin, a cholesterol synthesis inhibitor, introduced to the clinic to treat hypercholesterolemia, used either as a single or in combination with TNF-alpha inhibits growth of MmB16 melanoma and L1210 leukemia cells. PMID- 7487366 TI - Effect of cyclosporine A (CsA), cyclophosphamide (Cy) and azathioprine (Aza) on the survival of Listeria innocua in mice. AB - Immunosuppressors reducing the defensive mechanisms of the organism make favourable conditions for the development of different infections, particularly for intracellular facultative bacteria such as Listeria. In this paper, the effects of different doses of cyclosporine A (CsA), cyclophosphamide (Cy) and azathioprine (Aza) on the survival of Listeria innocua, a nonpathogenic strain, in the peritoneal cavity and spleen of BALB/c mice, were investigated. It was found that CsA and Aza, applied on total doses: 50-250 micrograms/g or 20-100 micrograms/g respectively, before or on the day of bacteria injection, prolong their longer survival as a result of reduced elimination. In CsA treated animals the number of survived Listeria is higher in the spleen while in Aza treated animals--in the peritoneal cavity. The Cy effect appeared to be the weakest because only the immunosuppressive (300 micrograms/g) dose was able to significantly enhance the survival of L. innocua both in the peritoneal cavity and the spleen. The results are a warning for clinicians applying the immunosuppressor treatment. PMID- 7487367 TI - Effect of H2O2 on adriamycin-induced cytotoxicity in vitro. AB - Normal and malignant cell lines, sensitive and resistant to adriamycin, have been treated with H2O2 to look for the possibility of protection from the cytotoxic action of adriamycin on the cells. Pre-treatment of adriamycin-resistant cells with H2O2 partially protects them against the cytotoxicity of adriamycin. PMID- 7487368 TI - Lymphocyte phenotype studies of rheumatoid arthritis patients treated with methotrexate. AB - Seventy nine patients with rheumatoid arthritis (RA) were monitored for 12 months. Forty eight individuals were treated weekly with a low dose of methotrexate (MTX), and 31 received nonsteroid antiinflammatory drugs (NSAID) only. The mean total dose of MTX was 413 mg with a range from 250 to 735 mg. The phenotype of lymphocytes was analyzed using double immunofluorescence and a panel of monoclonal antibodies. The clinical status of 33 MTX-treated patients improved, in 9 neither clinical nor laboratory improvement was noticed. Six patients were withdrawn from the study because of adverse reactions. We observed no changes in the percentage of CD3, CD4, CD8 and CD25 positive lymphocytes in both MTX and NSAID-treated groups. However, the percentage of CD19 positive cells significantly decreased during 12 month observation (15.1 +/- 6.5% v.s. 10.2 +/- 5.0, p < 0.01) in MTX-treated subjects. Moreover, this percentage increased (9.4 +/- 6.7% v.s. 17.1 +/- 1.1%, p < 0.01) in NSAID-treated patients. PMID- 7487369 TI - Contrasting effect of oral and intravenous cyclophosphamide treatment on phenotypes of human peripheral blood lymphocytes. AB - Twenty three rheumatoid arthritis (RA) patients treated with cyclophosphamide (CTX) were observed for 6 months. Eight patients received CTX in a single intravenous dose (group I) and 15 orally in a single daily dose (group II). The surface antigens of lymphocytes, isolated from peripheral blood, were determined using immunofluorescence method. The clinical improvement was observed both in group I and in group II of patients. However, in patients receiving CTX intravenously amelioration of the disease appeared quicker. The percentage of CD3+ and CD8+ T cells remained unchanged. In group I we observed decrease in the number of CD4+ T cells (60.1 +/- 11.5% and 43.8 +/- 12.5%, before and after treatment respectively, p < 0.01), in group II this level remained unchanged. In both groups the percentage of CD19+ B cells decreased (14.7 +/- 9.2% and 8.0 +/- 6.1%, before and after treatment respectively in group I, p < 0.01; 17.4 +/- 12.3% and 11.0 +/- 7.1%, before and after treatment respectively in group II, p < 0.01). Moreover, the percentage of activated T cells (CD25+ cells and HLA-DR+ cells) was reduced in both groups. PMID- 7487370 TI - Influence of various forms of dialyzable leukocyte extracts on rat adjuvant arthritis. AB - Adjuvant-induced arthritis in rats is a chronic inflammatory disease, widely used as an animal model for rheumatoid arthritis. In our study the effect of various fractions of dialyzable leukocyte extract (DLE): DLE I-molecular weight below 10 kDa (commercial preparation), DLE II-molecular weight below 5 kDa (suppressor fraction), DLE III-molecular weight 5-10 kDa on rat adjuvant-induced arthritis was studied. The adjuvant arthritic (AA) rats were treated with DLE fractions i.p. in solutions containing an active substance isolated from 12.5 x 10(6) and 6.25 x 10(6) leukocytes from day 1 (adjuvant injected) through day 18, every second day (total 9 times). Various markers of inflammation, immune function and joint destruction were evaluated: hindpaw volume, serum hyaluronic acid, serum albumin and biopterin in urine. All these markers showed a significant improvement after using fraction DLE II in comparison with AA controls. Fractions DLE I and DLE III influenced only some markers of inflammation and immune function. Our results demonstrated a therapeutical effect of fraction DLE II on rat adjuvant-induced arthritis. PMID- 7487372 TI - Influence of the perinatal period on bovine interferon production. AB - Interferon (IFN) production in cultures of leukocytes of calves immediately after birth, 2--3-day-old calves, dams and nonpregnant lactating cows was compared. For IFN induction Radom strain of Newcastle disease virus (NDV-R), phytohemagglutinin (PHA) and concanavalin A (ConA) were used. Cultures of newborn-calf peripheral blood mononuclear cells (PBMC) separated from blood by Ficoll-Hypaque density centrifugation or leukocytes cultivated by whole blood technique responded to all inducers used with a significantly lower IFN level than those of 2--3-day-old calves, dams and lactating cows. Although the mechanism of this phenomenon is not clear, our data connot rule out a defect of IFN synthesis in blood leukocytes of newborn calves. Spontaneous antiviral activity was detected in fetal membranes and in colostral leukocytes. Colostral leukocytes were efficient producers of IFN after induction with NDV-R, but less efficient than blood leukocytes after induction with PHA and ConA. Low response to PHA and ConA correlated with low number of lymphocytes in colostrum. PMID- 7487371 TI - Changes in the ultrastructure and functions of immunocompetent cells under the influence of external heating, interleukin 1 beta and leukocytic pyrogen. AB - The structural and functional organization of the rabbit immunocompetent cells after injection of human recombinant interleukin 1 beta (IL-1 beta), partially purified leukocytic pyrogen (LP), or after external heating (36 degrees C, 1.5 h) was compared. A rectal temperature increase being the same, there were similar and transient changes in epithelial and endothelial cells and macrophages, a rise in the proliferative activity of thymocytes and synthesis of IL-1-like activity, and a decrease in the interleukin 2 (IL-2) production by cultured splenic cells. PMID- 7487373 TI - Effect of macrophage-modulating agents on in vivo growth of transplantable Lewis lung cancer in mice. AB - C57Bl/6 mice, bearing transplantable Lewis lung cancer (non-metastatic subline) implanted either subcutaneously or intraperitoneally were treated with macrophage colony stimulating factor (M-CSF, 10(6) units per mouse, per day for 19 days), Escherichia coli lipopolysaccharide or both. Lipopolysaccharide (5 micrograms per mouse) administered daily once a day for up to 30 days impaired both subcutaneous and intraperitoneal tumor growth and prolonged survival of tumor bearing mice. Macrophage colony stimulating factor, administered daily, inhibited only subcutaneous tumor growth, both when administered alone and in combination with with lipopolysaccharide, and had no effect on intraperitoneal tumor. Moreover, it did not prolong survival of tumor bearing mice, when administered alone, and nullified the effects of lipopolysaccharide when administered concomitantly. These data suggest that macrophage colony stimulating factor, at least in this tumor model and in this dose schedule, offers little benefit. In contrast, the present data confirm earlier suggestions on therapeutic usefulness of bacterial lipopolysaccharide in neoplastic disease, which makes this compound an interesting candidate for future clinical trials. PMID- 7487374 TI - Cytotoxic T lymphocyte response to herpes simplex virus type 1 is composed of both CD8+ and CD4+ T cell phenotypes in acute and memory states. AB - Mice were infected via the cornea with HSV-1. Next, draining lymph nodes (DLN) and spleen cells were analyzed at various times post infection for the presence of cytotoxic T lymphocyte precursors (CTL-p) of both the CD8+ and CD4+ phenotypes. Responses were greatest in the DLN, but memory CTL persisted in the spleen and were undetectable in DLN by 60 days. On all occasions, the frequency of CD8+ CTL outnumbered CD4+ CTL. The murine CTL responses to HSV-1 differ from those in man where CD4+ MHC class II restricted CTL appear to dominate the response at least in the memory phase. PMID- 7487375 TI - Effect of carbon tetrachloride intoxication and ethanol ingestion on interferon production in mice. AB - Sixty C3H/He male mice were divided into 6 groups (10 mice per group). Control mice (group I) received three injections of PBS and drinking water. Mice of group II were injected with PBS but drinking water was substituted by ethanol solutions with increasing concentration of ethanol (w/v): 6% during the first week of experiment, 10% during the second and 20% during the third week. Group III received three intraperitoneal injections of CCl4 (1 ml of CCl4 per 1 kg of body weight) and water for drinking. Group IV was treated with CCl4 as group III, but drinking water was substituted by ethanol solutions with increasing concentrations as in the group II. Samples of blood, liver and spleen were taken 24 h after the third acute CCl4 intoxication. Group IIIa and IVa were treated as group III and IV, respectively, but samples of blood and organs were taken a week after the last CCl4 injection. A typical increase in serum ALT and necrosis of hepatocytes as confirmed by the histological examination, was observed 24 h after CCl4 injections (group III and IV). A week later (group IIIa and IVa) regenerative changes in the liver and a significant decrease in ALT serum activity was observed. Acute CCl4 intoxication (group III) significantly decreased IFN production in liver and spleen cells isolated 24 h after the last CCl4 injection. Combined CCl4 and ethanol administration affected very strongly IFN production (group IV).(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7487376 TI - N-acetyl-beta-D-glucosaminidase enzymuria as an indicator in monitoring the therapy of some rheumatic diseases with potentially nephrotoxic drugs. AB - The measurement of NAG enzymuria by the spectrofluorimetric assay according to Merle et al. enables an early diagnosis of tubular dysfunction. Urinary NAG activity was determined in 13 patients with various rheumatic diseases during administration of potentially nephrotoxic drugs like nonsteroidal anti inflammatory drugs (NSAID), aminoglycosides, gold salts, cyclosporin and steroids. The significant decrease of urinary NAG activity was common in patients with recently diagnosed rheumatic diseases, who received steroids. The decrease of NAG enzymuria was correlated with biochemical indices of inflammation like ESR or hemoglobin. The use of two types of potentially nephrotoxic drugs, like NSAID and gentamicin or NSAID with cyclosporin induced significant augmentation of NAG excretion. This occurrence may precede azotemia. The recognition of high NAG enzymuria permits to reduce dosage or discontinue treatment with potentially nephrotoxic drug prior to irreversible renal insufficiency as shown in the case of a patient with psoriatic arthritis treated with simultaneously administered cyclosporin and diclofenac. PMID- 7487377 TI - Opsonic activity of sera in children with hematological malignancies treated with Sandoglobulin. AB - Twenty seven children with hematological malignancies were treated with Sandoglobulin for life threatening infections due to severe granulocytopenia. We have studied the opsonic activity of sera in patients before and 7,14 and 21 days after the infusion of Sandoglobulin. Before the therapy a decrease of serum opsonic activity at the stage of ingestion and intracellular killing of bacteria has been shown. It was due to a deficiency of opsonizing factors. After treatment with Sandoglobulin the significant improvement of the opsonic activity of tested sera was found, but only at the stage of the ingestion of bacteria. The optimal interrelationship between opsonizing capacity of sera at the ingestion and intracellular killing phase was observed in the group of children treated with the relatively low Sandoglobulin dose (0.3-0.6 g/kg). In patients with the longest infection duration, who received the high Sandoglobulin doses (> 0.6 g/kg), the largest percentage of sera containing immune complexes was detected. These data demonstrate that high doses of globulins should be administered with certain care. PMID- 7487378 TI - College of American Pathologists Conference XXVIII on alternate site testing: introduction. AB - The College of American Pathologists Conference XXVIII, Alternate Site Testing Conference: Venturing Beyond the Boundaries of the Clinical Laboratory, was held from January 29 through February 1, 1995. This open format consensus conference gathered together individuals with many different points of view on the topic of hospital laboratory testing in noncentralized laboratories. The goals of the conference were to establish a basis for common beliefs about alternate site testing, to discover where consensus existed, and to establish future directions for consensus development and resolution of disputes in this sometimes contentious and confusing area of laboratory medicine. Attainment of the goals was attempted through a variety of conference activities, including plenary addresses, poster sessions, and, most importantly, through open-forum breakout sessions. During the breakout sessions all conference attendees were invited to contribute ideas toward the development of consensus about critical issues surrounding alternate site testing. Although it is clear that much work remains to be done if we are to learn how to use alternate site testing strategies to the best advantage, an important beginning has been made. The stage has been set as we continue to provide high-quality laboratory testing in an era that demands both fast turnaround time and optimal cost effectiveness. PMID- 7487379 TI - College of American Pathologists Conference XXVIII: Alternate Site Testing. January 29-February 1, 1995. PMID- 7487380 TI - College of American Pathologists Conference XXVIII on alternate site testing: keynote address. Reflections on a C major scale. PMID- 7487381 TI - College of American Pathologists Conference XXVIII on alternate site testing: keynote address. Global view of how alternate site testing fits in with medical care. PMID- 7487382 TI - Alternate site testing. The old and new paradigm or the past is prologue. AB - This paper reviews broad themes emerging in the discussion concerning alternate site testing, including the emergence and continuing value of centralized testing, the reasons for alternate site testing, and the relative merits and negative aspects of traditional centralized testing and decentralized models. The paper attempts to identify the expectations of constituencies affected by alternate site testing (patients, physicians, nurses, administrators and managers, and pathologists and other laboratory professionals) and identifies criteria and implementation strategies for decentralized testing. PMID- 7487383 TI - Responsibilities in point-of-care testing. An institutional perspective. AB - The development of "user-friendly" laboratory analyzers, combined with the need for rapid assessment of critical care patients, has led to the performance of in vitro diagnostic testing at the point of care. This strategy has been well received by most physicians who desire rapid turnaround times for laboratory tests, especially in the critical care areas. Since the primary care-giver in most critical care units is the registered nurse, much point-of-care testing has been delegated to nursing personnel. This has resulted in questions of authority, responsibility, regulation, and conflict resolution. These areas are discussed along with alternative strategies, such as vacuum transport, stat and/or satellite laboratories, unit-based phelebotomists, or "super techs," designed to achieve the goals of bedside or near-bedside testing in the critical care setting. PMID- 7487384 TI - Future practices in diagnostic medicine. PMID- 7487385 TI - Exploring the medical need for alternate site testing. A clinician's perspective. AB - The ability of the clinician to make rapid and precise decisions is the cornerstone of cost-effective patient care. Alternate site testing can provide rapid information to clinicians in emergency departments and critical care units. This can increase cost-effectiveness of treatment by shortening the length of hospital stays and reducing unnecessary treatment. Currently available alternate site coagulation methods allow for much more rapid achievement of effective and safe levels of anticoagulation. Potential alternate site methods of measurement of cardiac enzymes could reduce admissions to critical care units for chest pain and improve assessment of coronary artery reperfusion. PMID- 7487386 TI - Financial justification of alternate site testing. AB - Fixed costs are higher and variable costs lower for clinical laboratory testing than for alternate-site testing (AST). For AST to become cost-effective, variable costs must be reduced. Four models for accounting for the costs of clinical laboratory testing and AST are compared. A common error is to omit some of the indirect costs of a cost-per-test analysis for AST, thereby providing false claims. For a cost-per-test analysis, we recommend that identical categories be used for both the central laboratory and for AST. However, cost-per-test data alone do not completely represent institutional costs or savings associated with laboratory testing at any site. PMID- 7487388 TI - Alternate site testing. The decision-making process. PMID- 7487387 TI - Alternate site testing. The regulatory perspective. AB - This article provides a high-level overview of federal, state, and accrediting body requirements for laboratory testing outside the traditional laboratory environment. The Clinical Laboratory Improvement Amendments of 1988 provide a framework for most state and accrediting-body regulations. The stand taken by all regulators at this point is that testing wherever it is performed must conform to the same standards. The most variety in regulation is seen at the state level in the area of who can perform what tests and under what circumstances. All regulation focuses on the training and competency of the staff, adequate direction and supervision of the testing, and mechanisms for assessing the reliability of test results. PMID- 7487389 TI - Desirable performance standards for imprecision and bias in alternate sites. The views of laboratory professionals. AB - Many strategies exist for the delineation of desirable performance standards for imprecision and inaccuracy (bias). All have disadvantages and advantages. Currently, the professional consensus is that the favored approach is based on biology: desirable imprecision is less than one half of the within-subject biological variation and desirable inaccuracy is less than one quarter of the group (within-subject plus between-subject) biological variation. Current laboratory performance allows these goals to be met for many quantities. In the past, analyses done at sites other than the laboratory achieved poorer results, but recent advances in technology appear to allow some biology-based goals, which in our view are the appropriate standards, to be met. Further advances in setting and widely promulgating goals, making good calibrants widely available, and developing quality control, assessment, and management are all required to ensure that the desirable performance standards are obtained in all locations in the future. PMID- 7487390 TI - Reproducibility (precision) in alternate site testing. A clinician's perspective. AB - PROBLEM CONSIDERED: Reproducibility is only one dimension of test quality to be considered when evaluating the utility of a test or deciding to perform a test at or near the point of care. METHODS: Personal experience and review of the literature. RESULTS: Hemoglobin A1c and bedside glucose measurements are cited as examples in which a clinician may weigh the importance of precision differently. These examples are used to point out how precision should be considered in evaluating alternate site testing methodology. CONCLUSIONS: The importance of analytic precision to the clinician, and whether precision can be compromised for improvement of another dimension of test quality, depends on the context in which the result of the test is to be used. In addition to the purpose of the test, the level of precision required also depends on the patient population, the particular application of the test, and how the clinician interprets the test. Clinically relevant goals for test accuracy must be individualized for each test and application. Precision may be sacrificed for lower cost, increased availability, or faster turnaround time. Collaboration between laboratorians and clinicians is essential for optimal utilization of laboratory services. PMID- 7487391 TI - Timeliness of clinical laboratory tests. A discussion based on five College of American Pathologists Q-Probe studies. AB - Along with accuracy and reliability, timeliness is considered an essential quality for laboratory tests. Only for highly specific instances involving situations in operating theaters has timeliness been shown to affect outcome. Some studies have shown timeliness can shorten length of stay in certain emergency department situations, but rarely for hospital inpatients. Clinicians' expectations for emergency department laboratory test timeliness call for results in less time than the laboratory expects to or does provide. Data from five College of American Pathologists Q-Probes studies on test turnaround time support these observations, but also show central laboratories can provide results in a timely fashion if they optimize the total testing process. PMID- 7487392 TI - Alternate site testing. Information handling and reporting issues. PMID- 7487393 TI - Near patient blood glucose monitoring. AB - This report describes the quality control and quality assurance programs for bedside glucometry at our institution. From our 3-year experience, a regular schedule of inspections is necessary for maintenance of high-quality bedside glucose testing. The most common violation of quality control/quality assurance in bedside glucometry in our institution was the failure to perform regular proficiency testing, in which one random patient's bedside capillary blood glucose value during each inspection period was compared to his or her venous plasma or serum glucose value obtained from the central laboratory. Failure to perform instrument maintenance and document operator certification were the next most common violations of quality control/quality assurance in our bedside glucometry program. Regarding the cost analysis for bedside glucometry, we conclude that bedside glucose testing is not inherently more expensive than testing performed within the clinical laboratory. The increased cost of bedside glucometry over laboratory testing can be significantly minimized by involvement of a limited number of health care workers and performance of bedside glucometry only on clinical units where testing is required more than five times per day. PMID- 7487394 TI - Designing a point-of-care program for coagulation testing. AB - The prothrombin time and the activated partial thromboplastin time are routinely used to diagnose hemostatic disorders and to monitor anticoagulation therapy. Several coagulation test systems are now available that perform whole blood prothrombin time and activated partial thromboplastin time assays at the point of patient care. Many hospitals are evaluating point-of-care programs to satisfy the clinical demand for more expedient return of laboratory results. A successful point-of-care program requires the cooperation of laboratorians to assess validity of the methodology and clinicians to evaluate the clinical merit of the system. In this paper, the criteria for choosing a valid and appropriate point-of care coagulation system are discussed and guidelines for implementing a coagulation point-of-care program are suggested. PMID- 7487395 TI - The mobile laboratory in alternative site testing. AB - The establishment of mobile or portable laboratories as one strategy for delivery of laboratory services at alternative sites is evaluated. The mobile laboratory may be used to replace centralized laboratory testing in areas of critical need, such as critical care areas of the hospital in which relatively large numbers of tests are needed quickly. Other possible areas of use include outpatient clinics and other outreach settings in which care of the patient may be hastened by the availability of laboratory data on a real-time basis. In such areas where a need is established, mobile laboratory testing may be performed economically and may enhance the position of the medical technologist as a hands-on clinical caregiver. PMID- 7487396 TI - Successful alternatives to alternate site testing. Use of a pneumatic tube system to the central laboratory. AB - We evaluated the use of several modes of alternate site versus central laboratory testing in our hospital to provide improved processing of stat specimens from the Emergency Department and the Operating Room. Using the principle of parallel processing for sample analysis, coupled with installation of a computerized pneumatic tube specimen delivery system, we provided improved turnaround time, a larger test menu, and 24-hour-a-day service from the central laboratory. As a result, the satellite laboratory in the Emergency Department was closed and 16 full-time equivalent positions were saved in the hospital. A separate tube zone was established for the Operating Room, and a critical care whole-blood test unit was established in the central laboratory, providing 7-minute turnaround time for blood gases, electrolytes, and glucose. Testing in the Operating Room by nonlaboratory personnel was discontinued. PMID- 7487397 TI - Development of a robotic near patient testing laboratory. AB - Many laboratories have centralized critical care services to conserve resources, but centralization has often been at the expense of providing optimized patient care. We have created a remotely controlled clinical laboratory that provides whole blood analysis of blood gases (PCO2, PO2), pH, electrolytes (Na+, K+, Cl-), glucose, and hemoglobin near the patient's beside, while maintaining the distinct advantage of central laboratory control. The automated remote laboratory provides extremely rapid turnaround times, eliminates the cost of labor for specimen processing, reduces the risk from contaminated specimens, reduces staff training, and results in improved patient care. PMID- 7487398 TI - College of American Pathologists Conference XXVIII on alternate site testing: breakout sessions. PMID- 7487399 TI - College of American Pathologists Conference XXVIII on alternate site testing. What must we now do? AB - Implementation of the conference consensus that alternative site testing should be under the direction of the laboratory director supported by an institutional structure of users, laboratorians, and possibly administrators will optimize success. Information is needed if, when, and where alternative site testing improves the care of patients, and if faster results are associated with better and less expensive patient care. Costs and the timeliness of both central clinical laboratory and alternative site testing must be estimated in an identical fashion, then both must be compared with the costs and timeliness attributed to all steps of the total testing process included. As has been done with glucose, accuracy and precision goals must be developed for analytes measured at alternative test sites, then agreement reached on what these goals are and whether speed of measurement can influence these goals. Consideration of successful models is essential by those implementing or reevaluating alternative site testing programs. Required scientific information cannot be obtained in a timely manner unless improved funding occurs. PMID- 7487400 TI - The confidential unit exclusion and allogeneic blood donation. Results from supplementary questions to the Donor Center and Viral Markers surveys of the College of American Pathologists. AB - OBJECTIVE: The confidential unit exclusion (CUE) is a mechanism for allogeneic blood donors to confidentially indicate whether they feel their blood is safe for transfusion to others. The purpose of this national survey was to determine the extent of use of the CUE procedure and its related policies. DESIGN: Supplementary questions related to the use of the CUE were asked of those 1994 DC C, W1-C, and W2-C College of American Pathologists survey subscribers who collect allogeneic blood for transfusion. SETTING: National survey. PARTICIPANTS: Blood collection facilities who collect allogeneic blood components. MAIN OUTCOME MEASURES: Survey responses. RESULTS: There were a total of 2966 total participants in the 1994 DC-C, W1-C, and W2-C surveys. A potential total of 444 participants indicated that they collected allogeneic blood and were eligible to participate in the CUE survey. A variety of approaches are used concerning donor deferral, reinstatement, and follow-up in the use of the CUE. Documentation and donor counseling issues also show variation. CONCLUSIONS: Different approaches and procedures have been developed by collection facilities to address issues related to the use of the CUE. More data on the efficacy and cost-effectiveness of the CUE are needed. PMID- 7487401 TI - Practice protocol for the examination of specimens removed from patients with ovarian tumors. A basis for checklists. Cancer Committee, College of American Pathologists. Task Force on the Examination of Specimens Removed from Patients with Ovarian Tumors. PMID- 7487402 TI - Making the diagnosis of mixed and non-Alzheimer's dementias. AB - Community pathologists are often called on to perform autopsies to confirm clinical diagnoses of Alzheimer's disease, by far the most common cause of dementia. Diagnostic criteria have been provided by the Consortium to Establish a Registry for Alzheimer's Disease. Beyond pure and simple Alzheimer's disease, a significant proportion of dementia brains will feature Alzheimer's disease mixed with Lewy bodies, historically associated with idiopathic Parkinson's disease, or combined with various manifestations of cerebrovascular disease. Less commonly, the pathologist will encounter Lewy body disease alone, pure cerebrovascular disease, Pick's disease, progressive supranuclear palsy, Creutzfeldt-Jakob disease, or dementia lacking distinctive histopathology. This article is intended to reacquaint pathologists with these disorders and to provide a practical step by-step approach to making the diagnosis of these mixed and non-Alzheimer's dementias. PMID- 7487403 TI - The Kleihauer-Betke test. Clinical utility, indication, and correlation in patients with placental abruption and cocaine use. AB - This study examines the indications for performing the Kleihauer-Betke (KB) test and makes recommendations for its use. Results of 523 KB tests performed during 1993 at our hospital (Cleveland, Ohio) are reviewed in conjunction with surgical pathology reports of placental findings, obstetric records, and toxicology results. We conclude that the KB test should be performed following a positive screening test on all Rh negative mothers of Rh positive infants. Additional indications include cases of maternal trauma, unexplained increased maternal alpha-fetoprotein levels, fetal distress with abnormal heart tracings, intrauterine fetal death, and cases of unexplained neonatal anemia. We note that the KB test should not be performed to detect suspected placental abruption. PMID- 7487404 TI - Detection and monitoring of a concomitant atypical myeloproliferative disorder and chronic lymphocytic leukemia by flow-cytometric immunophenotyping. AB - OBJECTIVE: To illustrate the utility of a broad panel of monoclonal antibodies to detect secondary processes or unexpected characteristics of the primary blood dyscrasia. DESIGN: Case report and discussion. SETTING: Regional academic medical center. PATIENT: A 64-year-old man presenting with an apparent acute myeloid leukemia. INTERVENTIONS: Sequential immunophenotyping with a broad panel of monoclonal antibodies to monitor progression of disease and response to therapy. MAIN OUTCOME MEASURE: Identification and monitoring of the two atypical populations in this patient with correlation to the clinical status of the patient. RESULTS: Identification of an unsuspected mature lymphoid clone and characterization of the evolution of the myelomonocytic clone. CONCLUSION: The evolving mature lymphoid clone may have been overlooked in the context of a predominant atypical myeloproliferative process, particularly if a limited panel of monoclonal antibodies had been used for immunophenotyping. Sequential immunophenotyping was useful in monitoring the progression of each atypical process. PMID- 7487405 TI - Malignant mixed mullerian tumor of the extraovarian secondary mullerian system. Report of two cases and review of the English literature. AB - OBJECTIVE: To report two cases of malignant mixed mullerian tumor of the extraovarian secondary mullerian system and to identify cases reported in the English literature. DESIGN: Two cases are described and discussed along with cases reported previously in the English literature. SETTING: The University of Cincinnati Medical Center. PATIENTS: (1) A 62-year-old woman with a bilateral ovarian poorly differentiated endometrioid adenocarcinoma with multiple peritoneal metastasis and a malignant mixed mullerian tumor with heterologous elements arising from the pelvic peritoneum. (2) An 83-year-old woman with a malignant mixed mullerian tumor with heterologous elements arising from the cecal peritoneum. RESULTS: Seventeen previously reported cases were identified in the English literature. CONCLUSIONS: The malignant mixed mullerian tumor of the extraovarian secondary mullerian system is a rare disease with only 17 cases reported to date to our knowledge. The prognosis is poor. Of 12 patients with follow-up information available, 10 died within 1 year and 2 within 2 years after diagnosis. The histogenesis of the tumor remains unclear; however, data exist that support transformation of epithelial neoplastic cells into sarcomatous cells (metaplastic theory) and origin from a single totipotential cell. It is possible that both situations may occur. PMID- 7487406 TI - Microvascular invasion in prostate cancer correlates with pathologic stage. AB - OBJECTIVE: To determine the incidence of microvascular invasion in prostatic carcinoma and its correlation with other prognostic factors. DESIGN: Evaluation of the frequency and extent of microvascular invasion by routine light microscopy in 210 whole-mounted radical prostatectomies with cancer. RESULTS: Microvascular invasion was identified in 111 (53%) of 210 specimens. Focal microvascular invasion (involvement of fewer than three high-power fields) was present in 42 cases (20%); extensive microvascular invasion (involvement of three or more high power fields) was present in 69 cases (33%). Capsular perforation was present in 43% and 62% of cases with focal and extensive microvascular invasion, respectively. Seminal vesical involvement was observed in 23% and 47% of cases with focal and extensive microvascular invasion, respectively. Lymph node metastases were only observed in cases with microvascular invasion and were present in 7% and 23% of cases with focal and extensive invasion, respectively. There was also a strong positive correlation of microvascular invasion with surgical margin status, Gleason score, and cancer volume. Tumors ranged in volume from 0.14 cc to 47.25 cc (mean, 9.09 cc), and the volume in cases without microvascular invasion was lower than in those with invasion (mean, 5.4 cc and 12.7 cc, respectively); also, the volume in cases with focal invasion was lower than in those with extensive invasion (mean, 9.4 cc and 16.1 cc, respectively). CONCLUSIONS: Microvascular invasion in prostatic carcinoma correlates positively with virtually all measures of pathologic stage. Its presence should be reported in all prostate specimens according to the Cancer Committee of the College of Americans Pathologists. PMID- 7487407 TI - Interstitial amyloid deposits in the pituitary gland. Morphometry, immunohistology, and correlation to diseases. AB - The prevalence and immunoreactivity of interstitial amyloid deposits of the pituitary glands of 109 consecutive autopsies of individuals over 84 years of age were studied using Congo red staining and antibodies directed against the major amyloid fibril proteins and pituitary hormones. In addition, the amount of interstitial amyloid formation was evaluated quantitatively and compared with all autopsy-related and clinical diagnoses available. Eighty-seven (80%) of the 109 cases exhibited interstitial amyloid deposits in the anterior lobe. All reacted immunohistochemically with antiamyloid lambda light chain and antiamyloid P component. Quantitative analysis in 62 cases revealed a mean volume percentage of interstitial amyloid in the anterior lobe of 0.56%. In statistical analysis, only two of the 25 diseases recorded were associated with interstitial amyloid: chronic obstructive pulmonary disease and non-insulin-dependent diabetes mellitus. The prevalence of chronic obstructive pulmonary disease correlated positively with the occurrence (chi 2; P < .02) as well as with the amount of amyloid (Wilcoxon; P < .04) in the pituitary. In addition, non-insulin-dependent diabetes mellitus was accompanied with higher amounts of interstitial amyloid than with all other disorders (Wilcoxon; P < .03). Until now, a correlation was proposed only between non-insulin-dependent diabetes mellitus and islet amyloidosis of the pancreas. PMID- 7487408 TI - Meningioangiomatosis. A clinicopathologic study including MIB1 immunoreactivity. AB - OBJECTIVE: To examine the etiology of and clinicopathologic findings associated with meningioangiomatosis and to evaluate MIB1 immunoreactivity (marker of cell proliferation) in this lesion. DESIGN: Retrospective surgical pathology series of three patients. SETTING: Tertiary referral center with a high volume of epilepsy surgery. PATIENTS: Individuals with meningioangiomatosis who underwent surgical resection of the lesion. RESULTS: Three patients aged 11, 14, and 32 years (two females, one male) comprise the study. All patients presented with a history of intractable seizures (2, 3, and 30 years duration). None of the patients had von Recklinghausen's disease. All three patients underwent gross total resection of the lesion, and postoperative seizure-free intervals range from 7 to 15 months. Histologically, the lesion is characterized by a proliferation of blood vessels and meningothelial cells arranged around vessels in the meninges, cortex, and underlying white matter. Psammoma body formation or dystrophic mineralization and gliosis of the intervening parenchyma was observed in all three cases. Bodian stains failed to demonstrate neurofibrillary tangles within intervening parenchymal neurons. Focal epithelial membrane antigen positivity was observed in two lesions and was absent in one. MIB1 staining was observed in two lesions (MIB1 index = 0.8 and 0.6) within the meningothelial cell nuclei and was absent in one case. CONCLUSIONS: Meningioangiomatosis is a rare malformative or hamartomatous lesion of the central nervous system characterized by a proliferation of vessels and perivascular cuffs of meningothelial cells. Absent MIB1 immunoreactivity or low MIB1 indices in the lesion support the clinical impression of a slow-growing lesion and further support a malformative or hamartomatous etiology. PMID- 7487409 TI - Angiomatoid features in fibrohistiocytic sarcomas. Immunohistochemical, ultrastructural, and clinical distinction from vascular neoplasms. AB - The typical example of malignant fibrous histiocytoma (MFH) or dermatofibrosarcoma protruberans (DFSP) does not require ancillary studies for diagnosis. However, hemorrhage with cystic change consisting of blood-filled spaces may closely mimic a vascular neoplasm. Eight fibrohistiocytic sarcomas exhibiting these angiomatoid features, initially mistaken for vascular neoplasms, were identified from personal consultation files and review of 157 consecutive sarcomas (1985 through 1993) at the University of California-(Davis) Medical Center. They included five MFH giant-cell-type sarcomas, two MFH angiomatoid-type sarcomas, and one DFSP. Immunohistochemical analysis of paraffin-embedded material showed vimentin diffuse positive, CD68 (KP-1) diffuse positive, and factor VIII negative in all eight sarcomas; actin HHF-45 focal positive in six, diffuse positive in one, and negative in one sarcoma; desmin focal positive in two and negative in six sarcomas; and S100 protein, cytokeratin AE1:AE3, cytokeratin 10.11, and EMA negative in all eight sarcomas. Electron microscopy of three tumors exhibited neoplastic cells with fibroblastic, myofibroblastic, and histiocytic features. Weibel-Palade bodies or neolumens diagnostic of vascular differentiation were absent. The clinical characteristics and behavior of these sarcomas reflect entities in the spectrum of fibrohistiocytic lineage (MFH subtypes and DFSP) rather than vascular neoplasms. Patients with deep, large, giant-cell-type MFHs did poorly (two of four patients died from disease at 8 and 25 months). Both patients with angiomatoid MFHs showed local recurrences from large incompletely excised head and neck lesions. One died of disease at 21 months and the other is free of disease 12 months following excision of a local metastasis to the opposite side of the neck. The patient with DFSP had an 18-cm locally recurrent scalp tumor that extended into bone. Immunohistochemical and ultrastructural confirmation of fibroblastic, myofibroblastic, and histiocytic lineage and exclusion of vascular differentiation help to establish the correct diagnosis in these fibrohistiocytic sarcomas with angiomatoid features. The clinicopathologic features of these eight cases reaffirm the practical utility of MFH and DFSP as diagnostic entities in the spectrum of fibrohistiocytic sarcomas. PMID- 7487410 TI - Water-clear cell adenoma of the parathyroid. A case report with immunohistochemistry and electron microscopy. AB - We report a water-clear cell adenoma of the parathyroid gland, a lesion which to our knowledge has not been described previously. Like its rare but well-described hyperplastic counterpart, water-clear cell hyperplasia, this adenoma is composed of cells with abundant foamy-to-granular cytoplasm and mild nuclear pleomorphism. The cells form glandular structures and cell nests separated by fine fibrovascular septae. The tumor cells stain positively with anti-parathyroid hormone and show characteristic glassy and flocculate material by electron microscopy. Unlike water-clear cell hyperplasia, water-clear cell adenoma is a solitary lesion that compresses the residual nonneoplastic parathyroid gland. PMID- 7487412 TI - Synovial sarcoma of the heart. Correlation with cytogenetic findings. AB - A primary sarcoma of the heart was submitted to cytogenetic studies, which showed a translocation t(X;18)(p11;q11.2). The translocation between X and 18 is a characteristic one in synovial sarcoma of the lower extremities. As the histological appearance was not helpful for further classification of this primary sarcoma, karyotypic analysis proved useful in confirming diagnosis of synovial sarcoma, which, to our knowledge, is an uncommon neoplasm in the heart. PMID- 7487411 TI - Expression of alpha-fetoprotein by ovarian Sertoli-Leydig cell tumors. Case report and review of the literature. AB - A 17-year-old girl had an ovarian Sertoli-Leydig cell tumor associated with elevation of the serum alpha-fetoprotein level. On microscopic examination, the tumor had a predominantly spindled cellular stroma containing open and solid tubules, focal clusters of hepatoid cells, and heterologous carcinoid. Alpha fetoprotein was demonstrated within the hepatoid cells by immunohistochemical techniques. After surgery, serum alpha-fetoprotein level fell to normal, and the patient is still free of tumor 8 months later. PMID- 7487413 TI - Comparison of Ewing's sarcoma of bone and peripheral neuroepithelioma. PMID- 7487414 TI - Gonadal mixed germ cell tumor combined with a large hemangiomatous lesion. PMID- 7487415 TI - Fetal weights and measurements. PMID- 7487416 TI - Water-clear cell 'adenoma'. A new entity in the pathology of primary hyperparathyroidism. PMID- 7487417 TI - How to know when data may mislead. PMID- 7487418 TI - Transfusion medicine monitoring practices. A study of the College of American Pathologists/Centers for Disease Control and Prevention Outcomes Working Group. AB - OBJECTIVE: To survey transfusion medicine practices in 1990, to determine the distribution of defects in the transfusion process, to examine the relationship between defects and complications, and to recommend improvements in the transfusion process. DESIGN: A mail survey that divided the transfusion process into 24 risk-prone steps and gathered defect rates on each step, along with incidence data for eight known complications of transfusions and other demographic information. SETTINGS: Hospitals, independent laboratories, and blood centers that provide transfusion medicine services. OTHER PARTICIPANTS: Respondents were 1365 participants in the College of American Pathologists 1991 Blood Bank Quality Assurance Survey. RESULTS: While processing 6.2 million units of blood and blood products, respondents reported detecting over 88,000 defects: 41% in the preanalytic phase of testing, 55% in the postanalytic phase, and only 4% in the analytic phase, the phase to which most monitoring efforts were devoted. A median of eight steps were actively monitored by survey participants overall, whereas 96 facilities sought defects in all 24 steps. CONCLUSIONS: Analysis of the data showed several monitoring steps provide similar information. Although monitoring of the transfusion process could not be linked with prevention of the complications studied, active surveillance does focus attention on defect-prone steps and allows testing of strategies to improve the transfusion process. We describe how defect detection systems may be improved. PMID- 7487419 TI - Quality control of peptide drugs. Chiral amino acid analysis versus standard for icatibant acetate. AB - A method of chiral amino acid analysis versus standard is presented. By treating the peptide sample and a chiral standard whose intrinsic chiral composition is known in the same way, i.e. simultaneous hydrolysation and analysis, exact corrections can be made for the sequence-related and many hydrolysation-related racemisations. The accuracy obtained in this way permits use of the results for quality control of the chiral purity of peptide drugs. Using the bradykinin antagonist Icatibant acetate (INNM) the method was validated with respect to its precision, accuracy, specificity, limit of detection, and robustness. PMID- 7487420 TI - Synthesis, antitumoral and antiviral evaluation of halo- and demethyl-yatein derivatives. AB - Several chloro- and iodo-lignanolides have been obtained by direct halogenation of aromatic rings from yatein and 4'-O-demethylyatein. They were assayed as antineoplastics, in order to check the influence in the activity of substitution in both aromatic rings. Although these compounds show a modest antineoplastic activity, it is far from that displayed by yatein and podophyllotoxin. These results confirm that demethylation and the introduction of halo substituents diminish the activity of lignans of the dibenzylbutyrolactone type. PMID- 7487421 TI - Development of a parenterally administrable hydrosol preparation of the "third generation platinum complex" [(+/-)-1,2-bis(4 fluorophenyl)ethylenediamine]dichloroplatinum(II). Part 1. Preparation and studies on the stability and antitumor activity. AB - The development of a galenical formulation for poorly water soluble dichloroplatinum(II) complexes suitable for the parenteral administration in cancer chemotherapy is described. The procedure, which we elaborated for [(+/-) 1,2-bis(4-fluorophenyl)ethylenediamine]dichloroplatinum(II) (rac-4F-PtCl2), is based on the reaction of a soluble diaquaplatinum(II) salt with sodium chloride in water in the presence of pluronic F 68 as stabilizer and results in a sufficiently stable colloidal solution (i.e. hydrosol). In contrast to the poorly water soluble synthetic rac-4F-PtCl2, which was ineffective towards the hormone sensitive MXT-M-3.2 breast cancer of the mouse, its hydrosol formulation proved to be highly active and was very well tolerated. PMID- 7487422 TI - Structure-activity relationships in a series of 8-substituted xanthines as bronchodilator and A1-adenosine receptor antagonists. AB - Four new derivatives of 8-piperazine ethyl xanthine were synthesized and their bronchospasmolytic activity and A1-adenosine affinity were studied. Their relaxant action in the tracheal muscle was lower than that of theophylline and that of theophylline derivatives substituted at the 7-position. Only compound 9, where the methyl group in the 1-position of the theophylline was substituted by an isobutyl group, shows a good affinity towards the A1-adenosine receptor. PMID- 7487423 TI - Structure-activity relationship studies of CNS agents, Part 25. 4,6 di(heteroaryl)-2-(N-methylpiperazino)pyrimidines as new, potent 5-HT2A receptor ligands: a verification of the topographic model. AB - A series of new 4,6-di(heteroaryl)pyrimidines containing an N-methylpiperazino group (6-13) or an ethylenediamine chain (15-20) in position 2 were synthesized and their 5-HT1A and 5-HT2A receptor affinities were determined. It was shown that the substituent effects on the 5-HT2A affinity are additive and could be described quantitatively. In a behavioral model it was also demonstrated that 6 11 are 5-HT2A receptor antagonists. The molecular modelling results suggested that the distances between the basic nitrogen atom and the two aromatic centers (d1 = 5.2-8.4 A, d2 = 5.7-8.5 A, and d3 = 4.6-7.3 A) define the molecular topography of the 5-HT2A receptor antagonists under study. PMID- 7487424 TI - Antifungal agents, Part 11. Biphenyl analogues of naftifine: synthesis and antifungal activities. AB - A series of naftifine analogues having the biphenyl instead of the naphthyl moiety have been synthesized in a search devoted to study bioanalogues of clinically efficacious antifungal agents. The new derivatives were tested against Candida albicans by the direct contact method. They were also assayed against Gram-positive and Gram-negative bacteria and against some isolates of plant pathogenic fungi. Derivatives 8a, 8c, and 9a were found to be active against Candida albicans, derivative 5a was active against E. coli, a very resistant species to antimycotic agents, and derivatives 8a and 8b inhibited the plant pathogenic Rhizoctonia solani. PMID- 7487425 TI - Chemotactic peptide analogues. Synthesis and chemotactic activity of N-formyl-Met Leu-Phe analogues containing (S)-phenylalaninol derivatives. AB - The synthesis and the biological activity towards human neutrophils of some N formyl-Met-Leu-Phe-OMe analogues containing (S)-phenylalaninol (Pheol) or its derivatives in place of the native phenylalanine are reported. While the analogue containing Pheol (4) was found to be devoid of significant biological activity, its esters 3 and 5, although inactive as chemoattractants, are able to strongly stimulate superoxide production and are active with a lower efficacy in the lysozyme release. PMID- 7487427 TI - Consumer satisfaction with a rehabilitation mobile outreach program. AB - Accessibility to rehabilitation services is often difficult for people with disabilities who live in rural areas. This study examined consumer satisfaction with a rehabilitation outreach approach that utilizes a mobile clinic to provide consultation services. The descriptive survey took place in fifteen rural communities in eastern and northeastern Ontario, Canada. Valid postconsultation mail surveys were completed by 143 consecutive patients with mixed diagnoses (or proxy family members) who had been seen during mobile clinic visits to their home communities (85.1% of patients approached). There were 59 men and 84 women, with an average age of 58.7 years. The main outcome measure was a consumer satisfaction scale. A high level of global satisfaction was reported, with 97.2% of respondents reporting a preference for community outreach over the alternative of traveling to an urban rehabilitation center. Enhanced accessibility was considered to be the major advantage of the outreach program, but concerns about the continuity of care were also expressed. Providing interdisciplinary rehabilitation consultation services on an outreach basis is associated with a high level of consumer satisfaction. From a consumer perspective, the outreach approach seems to be a viable way of addressing some of the rehabilitation needs of rural people with disabilities. PMID- 7487426 TI - Non-steroidal anti-inflammatory agents, Part 19. E-2-pyrrolizin-5-yl acrylic acids as potent dual or selective inhibitors of bovine cyclooxygenase and 5 lipoxygenase. AB - The pyrrolizinyl substituted acrylic acid derivatives represent another class of dual and selective inhibitors of cyclooxygenase and 5-lipoxygenase. By modifying their substitution pattern at the phenyl moiety of C-6 the balance between the activity against cyclooxygenase and against 5-lipoxygenase can be shifted. Structure-activity relationships are discussed. Compound 6k is the most potent and well-balanced dual inhibitor of both enzymes, while the highest selectivity of lipoxygenase inhibition was found for 6j. The activity and selectivity of compounds with an additional sulfur moiety depend on the oxidation status of this atom, giving an indication of the discussed coupling between peroxidase and cyclooxygenase. The inhibition of cyclooxygenase was determined in a bovine thrombocyte intact cell assay and that of 5-lipoxygenase using intact bovine PMNLs. PMID- 7487428 TI - Measuring fatigue related to facial muscle function. AB - OBJECTIVE: The purpose of this study was to explore the expression of facial muscle fatigue in individuals without impaired muscle function using surface electromyography (EMG). DESIGN: Descriptive study of the expression of facial muscle fatigue in individuals without impaired muscle function. PARTICIPANTS: Convenience sample. Twenty individuals, 5 men and 15 women, between 20 and 50 years of age who volunteered to participate. OUTCOME MEASURES: Two tests of fatigue, a 10-second sustained contraction test, and a 25 repeated 3-second contractions test, were conducted on three facial expressions: brow raise, smile, and pucker. Surface EMG quantification of the muscle activity of the voluntary maximal facial muscle contractions was recorded during the fatigue tests. RESULTS: For the sustained fatigue test, all three expressions had a significant decline in activity (brow raise 34.51%, smile 22.96%, and pucker 29.05%); confirmed by a one-way ANOVA with repeated measures (brow raise df = 2, 38; f = 53.28; p = 0.00; smile df = 2, 38; f = 39.913; p = 0.00; pucker df = 2, 38; f = 76.002; p = 0.00). For the repeated fatigue test, percent fatigue was significant for smile (11.62%; df = 1, 19; f = 13.823; p = 0.001) but not for brow raise (7.27%; df = 1, 19; f = 1.945; p = 0.179) or pucker (4.22%; df = 1, 19; f = 2.508; p = 1.30). CONCLUSIONS: The muscle activity of sustained maximal voluntary muscle contractions of facial muscles fatigues significantly with time for brow raise, smile, and pucker expressions. The same facial muscles are more resistant to fatigue of muscle activity with repeated, brief contractions. Knowing the amount of facial muscle fatigue of individuals without impairment can be beneficial in developing outcome measures and goals for rehabilitation of individuals with facial neuromuscular dysfunction. Changes in fatigue tests of an individual with facial neuromuscular dysfunction with rehabilitation is reviewed for comparison. PMID- 7487429 TI - Comparing neuropsychological and psychiatric evaluation of competency in rehabilitation: a case example. AB - This report describes the case of a 20-year-old man who sustained a gunshot wound to the forehead, resulting in traumatic brain injury and C2 ventilator-dependent quadriplegia. Issues of personal control and autonomy typified his psychological adjustment. The question of competency to refuse medical treatment arose when he disallowed intervention for a suspected ear infection not confirmed by culture. Subsequently, the patient was alternately described as incompetent by a psychiatrist and competent by a medical ethics review panel. These decisions are interpreted within the context of existing medical-legal literature and historical precedent of competency in civil law. Central to competency evaluation is the patient's ability to recognize that a decision-making process is required, to review the pros and cons of various options, and to communicate a decision. Importantly, this decision need not be in accordance with the opinion of family or the health care team. Neuropsychological screening indicated the patient's cognitive abilities were within functional limits, and he subsequently agreed to treatment after experiencing pain and fever, and learning of a positive culture. It is concluded that a two-pronged neuropsychological evaluation of competency based on the patient's information processing capabilities is most appropriate in medical rehabilitation settings. PMID- 7487430 TI - The role of children in reporting their physical disability. AB - OBJECTIVE: To explore the contributions of children during the development of a disability scale, and their competence using the new scale. DESIGN: A new self report measure of pediatric physical performance, the Activities Scale for Kids (ASK), was developed based on interviews and pilot testing with children. The ASK was then filled out by children on two occasions 2 weeks apart to assess the reliability of child self-report. Validity was assessed by comparison of interview data and ASK scores from children with similar data collected from their parents, and to clinician and family global ratings of disability. SETTING: The study was conducted at a pediatric tertiary care hospital and its affiliated rehabilitation center. PATIENTS: All subjects, 5 to 15 years of age, experienced activity limitations because of musculoskeletal disorders and were free of cognitive impairment. Thirty children (mean age, 11.5 years) participated in the development of the ASK, and 28 children (mean age, 11.4 years) participated in testing of reliability and validity. RESULTS: Children generated items similar to those generated by parents (85% agreement) and identified 10 items not obtained from parents or the literature. Children demonstrated excellent test-retest reliability (intraclass correlation coefficient [ICC] = .97) using the ASK, and their scores were highly concordant with parent-reported ASK scores (ICC = .96). Validity was ascertained by comparison of ASK scores across different levels of disability based on global ratings of families and clinicians (p = .0023). CONCLUSION: Children are able to play an important role in pediatric physical disability evaluation. PMID- 7487431 TI - Self-managed versus agency-provided personal assistance care for individuals with high level tetraplegia. AB - OBJECTIVE: To determine whether a finite population, with severe physical disability, had better perceived quality of life, health status, and lower cost depending on whether they used agencies for their caregivers or hired, trained, and reimbursed their caregivers, independently. DESIGN AND SETTING: A survey, including demographics and portions of the Rand-36, LSI-A, PASI, PIP and CHART. The interviews were completed by telephone, by the leading author. PARTICIPANTS: Seventy-one persons who had sustained spinal cord lesions between C1 and C4. All were at least 1 year postinjury and had received rehabilitation at a local hospital in Englewood, Colorado. Ten additional persons came from the leading author's case management caseload or from a Boston rehabilitation center. They were interviewed in person for pretesting. MAIN OUTCOME MEASURES: Chi square, t tests, and multiple regression analysis, which controlled for potentially confounding group differences. Individuals were placed in two cohorts. Twenty nine persons received primarily agency provided care and 42 individuals used personal care attendants. RESULTS: The self-managed group showed significantly better health outcomes, with fewer rehospitalizations and diminished preventable complications. They also experienced greater life satisfaction and significantly lower costs. Those in the self-managed care group had significantly higher composite scores on Rand-36, indicating higher perception of recent health status. Significant differences in mean PIP scores indicated that members of the self-managed care cohort perceived greater control over day-to-day lives. PASI scores showed that persons in this cohort had greater satisfaction with their caregivers and costs. Cost benefit analysis showed appreciable lowering of costs in the self-managed care group. CONCLUSIONS: The self-managed care group had many more hours of paid attendant care, whereas the agency provider group had almost equal amounts of care from paid and unpaid providers. Persons using self-managed care reported having fewer medical problems, fewer hospitalizations and better perception of health. Participants declared greater satisfaction in having a choice of caregivers. Persons who were self-managing their care spent less money and used more hours of paid care. The financial burden borne both by the individual and society and the emotional burden borne by families and friends were diminished by the persons managing their care individually. PMID- 7487432 TI - Rightward orienting bias, wheelchair maneuvering, and fall risk. AB - OBJECTIVE: To investigate whether rightward orienting bias, without neglect of left hemispace, increased accident risk. DESIGN: Case-control study. SETTING: Inpatient rehabilitation unit of department of government medical center. PATIENTS: Successive right-cerebrovascular accident (CVA) admissions were reviewed over a 2-year period. Only patients with left hemisphere damage, recent abusive drinking, dementia, or inpatient stays of less than 3 weeks were excluded. Fifty-five right-CVA subjects were divided into three groups, based on starting point and omissions in left hemispace on the Rey-Osterreith Complex Figure Drawing and Random Letter Cancellation test. Thirty-two patients were placed in the L-OMIT group (omitted stimuli in left hemispace), 11 patients were placed in the R-BIAS group (began tasks to the right without omissions), and 12 patients were placed in the Non-Neglect Stroke group (no evidence of unilateral neglect). Twenty male inpatients with no history of brain damage served as controls (Normal Control). MAIN OUTCOME MEASURES: Frequency of hospital falls and wheelchair obstacle course contacts. RESULTS: Both the L-OMIT and the R-BIAS groups had more inpatient falls than the other groups F(3,71) = 6.11, p < .001. On the wheelchair obstacle course, the L-OMIT group made more left-sided wheelchair collisions than any other group. However, the R-BIAS group also made more errors than the Non-Neglect Stroke and the Normal Control groups, F(3,55) = 5.72, p < .01). CONCLUSIONS: Results suggest that rightward orienting bias has clinical significance, even without more serious symptoms of unilateral neglect. PMID- 7487433 TI - Mechanical and fatigue properties of wrist flexor muscles during repetitive contractions after cervical spinal cord injury. AB - OBJECTIVES: Force generation and fatigue properties of wrist flexor muscles were examined in subjects with chronic (> 1 year) cervical spinal cord injury (SCI, n = 16), and also in a control group of able-bodied (AB, n = 9) subjects. DESIGN: Using surface electrodes, wrist flexor muscles were stimulated with 126 trains of 26 stimuli at a frequency of 40Hz. The offset of each train was followed by a 1.5 second pause, for a total fatigue-test time of approximately 4.2 minutes. Isometric wrist flexion force was measured with a strain gauge. SETTING: This study was conducted at a research and rehabilitation center for spinal cord injury. MAIN OUTCOME MEASURES: Force profiles were analyzed for the maximum (peak) amplitude, the rise time, and the time constant of relaxation. RESULTS: At the outset, the average peak isometric measured in the SCI group was approximately one half that of the AB subjects. Although the relative decline in force with repeated stimulation was comparable between groups, the slowing of relaxation rate was much more pronounced in the SCI group. CONCLUSIONS: These findings are consistent with alterations in the metabolic profiles of wrist flexor muscles in the SCI group, probably reflected their altered activation pattern. When designing stimulation protocols for optimizing force and fatigue resistance in muscle left partially-paralyzed after spinal cord injury, particular care must be taken to allow adequate time for complete muscle relaxation, to avoid overdriving of the muscle and a loss of functional capacity. PMID- 7487434 TI - Rasch analysis for outcomes measures: some methodological considerations. AB - OBJECTIVE: To assess the four approaches to applying Rasch analysis to ordinal, repeated measures: (1) to perform separate Rasch analyses; (2) to perform a single analysis by treating the subjects measured on different occasions as distinct ones; (3) to perform a single analysis by treating items used on different occasions as distinct ones; and (4) to construct a 3-facet model by adding occasions as the third facet. DESIGN: Various Rasch and 3-facet models applied to retrospective, cohort data. SETTING: A stroke program at a rehabilitation hospital. PATIENTS: 99 of the 107 consecutive patients admitted between March and October, 1990, average age 69 years, 40% of whom were women, with an average stay of 54 days. INTERVENTION: Medical inpatient rehabilitation. MAIN OUTCOME MEASURE: The Functional Independence Measure (FIM), administered within 1 week of admission and discharge, and approximately 6 weeks after discharge. RESULTS: The four approaches resulted in various occasion-specific and generalized measures of item difficulty, patient ability, and occasion severity. Although these measures turned out to be congruent with each other as they should, they generated different sources of information needed for formative and summative evaluation of health care outcomes. All four approaches were found to be useful, although each had its own limitations. A four-step strategy based on these approaches was therefore designed and recommended. CONCLUSION: Rasch analysis of ordinal, repeated measures poses a special challenge to those interested in health care outcomes. A multi-method strategy incorporating various approaches is therefore needed to optimize its use in health care research. PMID- 7487435 TI - Decreased balance performance in cowboy boots compared with tennis shoes. AB - OBJECTIVE: This study was conducted to examine the relative balance performance of cowboy boots versus tennis shoes when the subject was challenged with different accelerations. The end point was the highest acceleration at which the subject could maintain full foot contact with the platform for each footwear type, ie, break acceleration. DESIGN: Crossover trial. SETTING: General community. PARTICIPANTS: Twenty-seven healthy women, 18 to 40 years old, with shoe sizes between 6 and 9, were selected in a convenience sample after their response to posted ads in a university medical center. INTERVENTION: Each subject was tested 3 times in the forward direction per acceleration on a Motionspec balance platform. A successful series was defined as keeping feet flat for 2 of 3 tests/level. MAIN OUTCOME MEASURE: The planned major outcome was the difference between the highest accelerations at which the subject was successful for each type of footwear. RESULTS: Subjects were found to have a highest sustainable acceleration in boots 10.66 +/- 6.20cm/s2 less than the highest acceleration in shoes. A period effect was found that improved the results of the second footwear tested by 7.2 +/- 6.20cm/s2. CONCLUSIONS: Cowboy boots have a decreased balance performance compared with tennis shoes. Further study should examine the specific features in the boot that contribute to this imbalance and examine the kinematic adaptations of the body to cowboy boots. PMID- 7487436 TI - Electromyography: do the diagnostic ends justify the means? AB - BACKGROUND: Physicians are sometimes reluctant to refer patients for electrodiagnostic studies (electromyography with nerve conduction studies [EMG/NCS]) believing the test is too painful and of little benefit. METHODS: We performed two separate surveys on 126 and 100 consecutive patients referred to our laboratory to determine if EMG/NCS was beneficial to the referring physician and to compare the level of anxiety experienced by patients before the study with the pain actually experienced during the study. RESULTS: The electrodiagnosis was discordant from the referring diagnosis in 39% of the patients with an abnormal EMG/NCS. Pretest anxiety levels were low in 59% of the patients, medium in 27%, and high in 14%. After the tests, 82% of the patients said that the test was not as bad as expected, and was generally only mildly painful. Ninety-three responded that they would have the test performed again. CONCLUSIONS: EMG/NCS often suggest alternative diagnoses, and the actual pain experienced during an EMG/NCS study is significantly less than expected. PMID- 7487437 TI - Employment after rehabilitation for musculoskeletal impairments: the impact of vocational rehabilitation and working on a trial basis. AB - OBJECTIVE: To investigate to what extent vocational rehabilitation and working on a trial basis have an impact on employment after rehabilitation. DESIGN: Follow up survey, conducted in 1991, among all patients, aged 14 to 64, who were treated between 1984 and 1987. SETTING: An out-clinic department of a rehabilitation center in the west of The Netherlands. SUBJECTS: 395 patients (59% of the original sample) participated in the study. They suffer from musculoskeletal impairments due to back pain (41%), brain injury (20%), neurological disorder (12%), trauma to the upper and/or lower extremities (8%), rheumatoid arthritis (7%), or other disorders (12%). No differences were found between those who did and those who did not respond with regard to age, gender, marital status, disorder, work experience, and participation in vocational rehabilitation. MAIN OUTCOME MEASURE: Odds ratio of having a paid job after rehabilitation. RESULTS: Participation in vocational rehabilitation and working on a trial basis depends on age, gender, work experience, and disorder. When statistically controlled for these variables vocational rehabilitation (odds ratio 1.96; 95% confidence interval 1.12 to 3.42) and working on a trial basis (odds ratio 3.26; 95% confidence interval 1.74 to 6.11) proved to have a significant impact on employment after rehabilitation. CONCLUSIONS: The findings suggest that rehabilitation programs that aim specifically at promoting employment for people with disabilities are effective, in particular when they take place in both a laboratory and a natural setting. The validity of this suggestion must be further tested by means of an experimental design. PMID- 7487438 TI - Community-dwelling older adults' attributions about falls. AB - OBJECTIVE: To examine the extent to which 162 older adults attributed the cause of their fall to their own limitations and to their surroundings, and the relationship of these attributions with selected sociodemographic, health related, locus of control, and fall-related characteristics. DESIGN: A stratified (by age > or = 65, gender, and area of residence) random sample; needs assessment survey on products and services to enhance independent living among older adults; included section on falls. SETTING: Community-dwelling individuals in both urban and rural settings. PARTICIPANTS: Potential respondents were drawn from the Manitoba Health database. Overall refusal rate was 22%. Final number of 1,406 participants were interviewed; 162 of these participants reported a fall. MAIN MEASURES: Outcome measures were: (1) the extent to which the fall was attributed to "own limitations" (internal attribution), and (2) the extent to which the fall was attributed to "surroundings" (external attribution). RESULTS: There is diversity in the extent to which individuals attribute falls to internal and external causes. Stepwise regression analyses indicated those reporting poorer self-rated health (p < .001); having dexterity difficulties (p < .01); and living in an apartment (p < .05) were more likely to attribute the fall to their own limitations. Those reporting better self-rated health (p < .01); and falling outdoors (p < .01) were more likely to attribute the fall to the surroundings. CONCLUSIONS: Fall attributions need to be more fully examined in the context of self-rated health and fall prevention programs. PMID- 7487440 TI - Randomized controlled trials in industrial low back pain relating to return to work. Part 1. Acute interventions. AB - Employers and insurers are interested in being able to use cost-effective interventions for returning injured workers to the workplace. Unfortunately, truly objective information is lacking. The purpose of this and two subsequent review articles was to perform thorough scrutiny and methodologic comparison among all obtainable, published randomized and controlled studies on low back pain (LBP) interventions leading to return to work. The study was confined to English language articles published from 1975 through 1993. Of more than 4,000 LBP citations, more than 500 were chosen for review. Of that number, 35 articles met the selection criteria of randomization, reasonable controls, and work return comparisons. This paper focuses on the 10 articles relating to interventions for acute (less than 4 weeks) LBP, and considers bed rest, exercise, spinal manipulation, back school, and case management. A 26-point quality system was used to compare the methodologic rigor of each article. This literature survey demonstrated the meager scientific foundations on which our industrial rehabilitation programs are based. PMID- 7487439 TI - Age and gender effects on postural control measures. AB - OBJECTIVE: Identifying age-related changes in the postural control system is an important first step towards understanding the risk for falls in older adults. The purpose of this study was to determine whether age or gender had any effects on six relatively new postural control measures. DESIGN: Cross-sectional, 2 x 2 factorial design, representing two ages (younger [20 to 35 years], and older [60 to 75 years]) and both sexes. SETTING: University campus. PARTICIPANTS: Twenty four community dwelling healthy adults (12 women, 12 men) participated in each of the younger and older groups. MAIN OUTCOME MEASURES: Outcome measures included five force platform measures and functional reach (cm). The force platform measures, obtained using the Balance Master system, included movement time and path length to targets, and sway area under conditions of eyes open, eyes closed, and with visual feedback. RESULTS: Although gender was not significant for any outcome measure, age was significant in all six outcome measures. Older adults demonstrated larger areas of sway regardless of condition (eyes open, eyes closed or with visual feedback). Older adults had longer movement times, longer path lengths, and shorter distances of functional reach when compared with younger adults. CONCLUSIONS: The results suggest that the measures studied are sensitive to age-related changes in healthy elderly. PMID- 7487441 TI - Breast cancer recurrence in a patient with a previous history of radiation injury of the brachial plexus: a case report. AB - Brachial plexus injury with resultant pain and attendant impairment may occur in cancer patients from either disease recurrence or as a consequence of radiation therapy. Distinguishing between these two causes of plexus dysfunction may be difficult, but physical findings and appropriate workup may assist in diagnosis. Additionally, lymphedema may be seen as a result of surgical, radiation, or combination therapy for breast cancer. Patients typically present with arm swelling, but neurological impairment, including pain, may also be noted. This case report describes an individual who developed pain and weakness, initially from radiation injury, and later developed local disease recurrence. PMID- 7487442 TI - Academic productivity. PMID- 7487443 TI - Acute and subacute rehabilitation. PMID- 7487444 TI - Lower extremity SEPs. PMID- 7487445 TI - Walking cadence effect on plantar pressures. AB - OBJECTIVE: Prior studies have examined the effect of cadence on ground reaction forces by use of a force plate. Force plate studies generally analyze isolated steps and do not provide insight into ongoing step-to-step variations or in-shoe plantar pressures. The objective of this study was to evaluate the effect of walking cadences on in-shoe plantar pressures over extended periods of continuous walking. DESIGN: Nonrandomized control trial. SETTING: Laboratory. PATIENTS OR OTHER PARTICIPANTS: Volunteer sample of 8 able-bodied subjects. INTERVENTIONS: In shoe plantar pressures were studied during four minutes of continuous walking at controlled cadences of 70, 80, 90, 100, 110, and 120steps/min. For each cadence more than 200 steps were analyzed for each of the 8 subjects. MAIN OUTCOME MEASURES: Pressure-time integrals, foot-to-floor contact durations, and peak pressures at all 14 locations were processed for each step. Changes were calculated compared to values at 70steps/min. RESULTS: With increasing cadence, mean pressure-time integrals continuously decreased (45% at 120steps/min); mean foot-to-floor contact durations continuously decreased (64% at 120steps/min); and mean peak pressures increased (119% at 120steps/min). CONCLUSIONS: Our results show that as walking cadence increases, pressure-time integrals and foot-to-floor contact durations decrease, and peak plantar pressures increase. This is clinically relevant to all kinetic gait studies because our results suggest that normal values should be established for each cadence. PMID- 7487446 TI - Transient traumatic quadriplegia in a 29-year-old man without predisposing anatomic lesion. AB - A 29-year-old man presented 45 minutes after sustaining a blow to the right side of his head when his motorcycle slid on gravel and flipped on the side of the road. After walking away from the accident, the patient noted burning pain in his neck. Shortly thereafter, complete quadriplegia with a sensory level at C4 developed. The patient was given a high dose of methylprednisolone and transferred to a tertiary care center. En route, neurological testing demonstrated rapid improvement, and on arrival, all abnormal motor and sensory findings had completely resolved. Radiographic examination and magnetic resonance imaging of the entire spine showed no abnormality. Torg's ratio was > 1 at vertebral level C3 through C6. There was no evidence of developmental stenosis, congenital fusion, cervical instability, or intervertebral disk protrusion. PMID- 7487447 TI - Lumbar-sacral radiculopathy secondary to intraspinal synovial cyst. AB - OBJECTIVE: The presentation of a patient with acute low back pain and distal radiation to the lower extremities is often attributed to a herniated nucleus pulposus (NHP). The purpose of this report is to illustrate how an intraspinal lumbar synovial cyst can have a similar presentation. CASE PRESENTATION: A 52 year-old man presented with low back pain with left lower extremity weakness and distal radiation. An electrodiagnostic evaluation was consistent with lumbar sacral radiculopathy. Computed tomography and magnetic resonance imaging showed a synovial cyst of the L4-5 facet joint. INTERVENTION: The patient underwent a L4 L5 laminotomy, synovial cyst excision, and decompression of the L5 nerve root. RESULTS: There were no postoperative complications. The patient had residual left lower extremity numbness but gradually regained the strength of his left lower extremity. Intraspinal synovial cyst can mimic the clinical pattern of NHP. CONCLUSION: An intraspinal lumbar synovial cyst can present with symptoms of nerve root compression. Given the presentation of lumbar-sacral radicular symptoms such as radiating pain, muscle weakness, and numbness, surgical excision of the lumbar synovial cyst remains the definitive treatment of choice. PMID- 7487448 TI - Home mechanical ventilation in mitochondrial encephalomyopathy syndrome. AB - Long-term home mechanical ventilation of children has only recently become more practically feasible and ethically acceptable by the medical community. It has been particularly controversial in cases of degenerative myopathies in which quality of life has been questioned. There are no reports in the literature of long-term home mechanical ventilation of a child with mitochondrial encephalomyopathy (MELAS) syndrome despite the many descriptions of possible etiologies of the concomitant respiratory failure. The patient reported here has used home mechanical ventilation for 6 years with few medical complications, no hospitalizations in the past 3 years, and increased function in activities of daily living. Despite the ill-defined nature of the disease and uncertain prognosis, we believe that long-term home mechanical ventilation of children with early onset MELAS syndrome is a viable option for both patients and their families and results in overall improvement in quality of life for the patient. PMID- 7487449 TI - Persistent dizziness following head trauma and perilymphatic fistula. AB - A growing body of evidence supports the idea that dizziness that persists for months and even years can be caused by an unsuspected perilymphatic fistula. Perilymphatic fistulas are abnormal ruptures that allow perilymph to leak out of the inner ear into the middle ear space. Most commonly, these ruptures occur secondary to a traumatic event. The term postconcussive syndrome has been used to describe a myriad of symptoms following head trauma. Some of these symptoms, such as cognitive changes, tinnitus, neck stiffness, and dizziness, are also commonly caused by active perilymphatic fistulas. This article discusses the typical history and diagnostic tests for patients with perilymphatic fistula. Common diagnostic tests include audiograms, electronystagmograms, electrocochleograms, and subjective and platform fistula tests. Also, the surgical treatment for the perilymphatic fistula (ie, repair of the oval and round windows) is reviewed, along with the results produced by this relatively minor ear operation. Suggestions are made to help the medical professionals involved in rehabilitative care to be aware of perilymphatic fistulas and seek proper consultations from inner ear specialists if they suspect the existence of this easily cured disorder. PMID- 7487450 TI - Decreased isokinetic trunk muscle strength and performance in long-term survivors of childhood malignancies: correlation with hormonal defects. AB - OBJECTIVE: To evaluate trunk muscle strength and performance in long-term survivors of childhood malignancies relative to age and sex-matched controls, and to relate the muscle strength observations to the therapeutic agents used and possible hormonal disturbances. DESIGN: Age and sex-matched cohort study. SETTING: Referral center in the northern part of Finland. PATIENTS: Forty-six long-term survivors of childhood cancer. Mean age at examination 19.1 years and median off-therapy time 9.4 years. INTERVENTION: Isokinetic dynamometer testing. MAIN OUTCOME MEASURES: Measurements of trunk muscle peak torque (PT) and total work done (TWD) were performed at angle speeds of 50 degrees/sec and 200 degrees/sec. The results were normalized relative to body fat-free weight (FFW). RESULTS: PT in the trunk muscles was lower in the patients at both angle speeds (mean normalized PT = 5.7Nm/kgFFW vs 7.6Nm/kgFFW for controls at 50 degrees/sec), as also was TWD except for extension TWD at the higher angle speed (mean normalized TWD = 59.9J/kgFFW vs 84.6J/kgFFW for controls at 200 degrees/sec). The normalized PT at 50 degrees/sec and TWD at 200 degrees/sec were lower in the males with testicular damage; also, low age at diagnosis correlated positively with muscle strength and performance. There were no differences in normalized PTs or TWDs between cranial radiation and non-radiation cases, or between growth hormone-deficient and non-deficient cases, and the patients without cranial radiation or with normal growth hormone secretion still had lower normalized PTs and TWDs than the controls. CONCLUSIONS: Survivors of childhood malignancies have decreased maximal trunk muscle strength and performance. The etiology of this effect remains unclear, but young age at diagnosis, as well as serum testosterone levels in male survivors, evidently influence muscle strength and performance. PMID- 7487452 TI - Standing balance and function over the course of acute rehabilitation. AB - The purpose of this study of 52 acute rehabilitation patients was to describe the reliability and responsiveness of measurements of standing balance and function and to establish the relationships between the measurements over the course of rehabilitation. Standing balance was measured with a seven-level (0 through 6) ordinal scale. Three functional activities (chair to mat transfer, locomotion, and stair climbing) were measured using Functional Independence Measure (FIM) scores. Balance and FIM scores were found to be reliable (weighted kappa > .85). Balance and FIM scores increased significantly (p < .000) between initial and final assessments (mean interval = 17.5 days). All FIM scores were correlated significantly, both cross-sectionally (rs = .444 to .774) and longitudinally (rs = .279 to .616), with balance. Moreover, changes in FIM scores were correlated significantly with changes in balance (rs = .413 to .595). The results of this study support the use of the ordinal balance scale described herein in an acute rehabilitation setting. PMID- 7487451 TI - Electromyographic and neuromuscular variables in post-polio subjects. AB - OBJECTIVE: Post-polio subjects experience functional deterioration many years after developing acute poliomyelitis and have been shown previously to have a deficit in strength recovery after isometric activity. This study characterized the size and stability of the motor units in a group of post-polio subjects with macro and single fiber electromyography (EMG) and correlated these variables with isometric strength, endurance, "work capacity," and strength recovery after fatiguing isometric exercise. DESIGN: A cohort of 12 post-polio subjects was tested for neuromuscular function. Electromyographic variables were determined on a separate day. SETTING: Volunteers were recruited from the community and tested in our neuromuscular research laboratory. SUBJECTS: A volunteer sample was obtained from advertisements. All subjects acknowledged post-polio syndrome symptoms. MAIN OUTCOME MEASURES: Neuromuscular variables were isometric knee extension peak torque, endurance (time to exhaustion) at 40% of maximal torque, tension time index, and recovery of torque at 10 minutes. Electromyographic variables were macro EMG and single fiber EMG (percent blocking and jitter). RESULTS: Macro EMG amplitude was ninefold the control value, and both jitter and blocking were greatly increased in comparison to control values. Isometric strength significantly (p < .05) correlated negatively with macro EMG amplitude. CONCLUSIONS: The weakest subjects had the greatest number of muscle fibers within the motor unit (as measured by macro EMG amplitude). Jitter and blocking did not correlate with neuromuscular function. PMID- 7487453 TI - Gait speed and activities of daily living function in geriatric patients. AB - OBJECTIVE: To establish the relationship between gait speed (GS) and functional independence in elderly people. DESIGN: GS is suggested as being a criterion standard in rehabilitation reflecting muscle strength. This study assessed the relationship between gait speed and functional independence in Activities of Daily Living (ADL). GS was measured by portable accelerometer over 2 meters. The mean of 3 attempts was taken. ADL function was measured by an occupational therapist using the modified Barthel ADL Index. The relationship between these measures was assessed by a statistician. SETTING: A geriatric unit in a hospital in Scotland. PATIENTS: One hundred sixty-one inpatients and outpatients were selected at random from the patients of a geriatric unit over a 3-month period. Patients were eligible if they were mobile with or without a walking aid. INTERVENTIONS: GS was measured by portable ultrasonic accelerometer. Patients were reviewed by an occupational therapist, blinded to their GS, who recorded functional capacity. Case sheet review provided diagnostic details and cognitive function. The type of floor surface was recorded. MAIN OUTCOME MEASURES: GS (m/sec), and Barthel score. RESULTS: Patients with GS of < .25m/sec were more likely to be dependent in one or more ADL function, p < .01. Those with a GS between .35 and .55m/sec were more likely to be independent in all ADL functions, p < .001. Patients whose GS was > .55m/sec did not maintain this independence. There was no relationship between GS and floor surface or cognitive function. CONCLUSIONS: GS is a useful indicator of ADL function in geriatric patients. PMID- 7487454 TI - The Annual Scientific Congress of the Royal Australasian College of Surgeons. AB - The Royal Australasian College of Surgeons (RACS) is responsible for the training, examination, and recertification of its fellows, who make up the great majority of the practicing specialist surgeons in Australia and New Zealand. One of the 16 Objects of the College is "To bring together the surgeons of Australia and New Zealand periodically for scientific discussion and practical demonstration of surgical subjects." In the furtherance of this object, there are many local and national meetings each year, but the highlight is the Annual Scientific Congress (ASC), held in each of the major cities on rotation, traditionally in the month of May, which is late autumn in the southern hemisphere. PMID- 7487455 TI - Managed care in JAMA and the Archives journals. A call for papers for coordinated theme issues. PMID- 7487456 TI - We hold these truths.... AB - It is traditional in an address of this kind in surgical organizations for a great social, national, professional issue to be analyzed, often quite correctly and brilliantly. A clarion call for a plan of action is outlined, actions that are succinctly and brilliantly captured. If you peruse most of the Presidential Addresses of surgical organizations, you will find that as a pattern. I am not certain I can do that very well. I am not sure I know what to tell you about the nation's and the world's problems. I am not sure that if I had an action plan, it would have any credibility or that anybody would follow it, however correct it might be. And so, rather than making pronouncements about the all too real issues that confront us, what I would like to talk about are some matters that underlie all that we do--the constants--those aspects that at least I perceive as truths underlying what we do. Some of you may argue that these matters have little or nothing to do with the scientific organization or scientific meeting and will not convey anything of value. I hope to prove you incorrect. I hope at the end of this brief talk that you will take something away from it. Certainly I am sincere in my offering. I am not trying to sound arrogant or accomplished, nor do I presume to know all the answers. These are merely observations and things I think about. PMID- 7487457 TI - Association of interleukin-4 plasma levels with traumatic injury and clinical course. AB - OBJECTIVES: To determine if interleukin-4 (IL-4) could be detected in plasma of trauma patients and if IL-4 activity is associated with patterns of clinical events, complications, or outcomes. METHODS: A prospective case series conducted in a tertiary care referral center with a level I trauma center. One hundred patients admitted to the trauma intensive care unit for at least 3 days were included. Plasma concentrations of IL-4 and IgE were determined from admission to intensive care unit discharge. Data on clinical outcome were collected, including death, sepsis, severe sepsis, adult respiratory distress syndrome, pneumonia, and renal dysfunction. RESULTS: Interleukin-4 was detected in the plasma of 87 patients. Patients with an Injury Severity Score of greater than 25 had higher admission IL-4 levels (P = .03) and greater maximal IL-4 levels (P < .001). Admission hypotension (P = .04) and age 30 years or younger (P < .001) were also associated with higher admission IL-4 levels. Increases in IL-4 levels were significantly greater for patients in whom sepsis, severe sepsis, or pneumonia developed (P < .05). A low admission IL-4 level was associated with a greater incidence of nosocomial pneumonia (P < .001). Additional indirect evidence of IL 4 activation included increased plasma IgE levels. CONCLUSIONS: Anti-inflammatory cytokine mechanisms are activated after injury and are associated with the development of infectious complications (sepsis, severe sepsis, and pneumonia). Exogenous administration of interleukin-4 should be evaluated as an experimental therapeutic approach after trauma and associated sepsis. PMID- 7487458 TI - Bombesin affects mucosal immunity and gut-associated lymphoid tissue in intravenously fed mice. AB - BACKGROUND: Our prior studies show that intravenous (IV) total parenteral nutrition (TPN) produces atrophy of the small intestine-related gut-associated lymphoid tissue and significant decreases in intestinal IgA levels, the major system of mucosal immunity. Others have noted increased small intestinal permeability, bacterial adherence and translocation, and decreased IgA levels in TPN-fed animals. Bombesin, a neuropeptide, may play a regulatory role in mucosal immunity. It is not clear whether bombesin attenuates the TPN-associated gut associated lymphoid tissue atrophy. OBJECTIVE: To examine the effect of bombesin on gut-associated lymphoid tissue integrity and function during IV TPN feeding. DESIGN: Randomized animal study. SETTING: A university laboratory. MATERIALS AND METHODS: Male ICR mice weighing 25 to 30 g were randomized to chow plus IV saline solution (n = 12), IV TPN (n = 12), or IV TPN plus bombesin (15 micrograms/kg, administered intramuscularly three times a day) (n = 12). Animals were killed after 5 days of receiving the experimental diet. Total small intestinal IgA level was quantified by enzyme-linked immunosorbent assay. Lymphocytes were isolated from Peyer's patches, intraepithelial spaces, and lamina propria and were stained with specific antibodies for B and T cells and for T-cell expression of CD4 and CD8 by flow cytometric analysis. Data were analyzed by analysis of variance. RESULTS: Bombesin prevented the IV TPN decreases in (1) total cell yield and B cell yield from the Peyer's patches, intraepithelial spaces, and lamina propria; (2) T-cell yield in the intraepithelial spaces and lamina propria; and (3) small intestinal IgA levels. Bombesin also reversed IV TPN decreases in CD4+ and CD8+ T cells in the intraepithelial spaces and Peyer's patches and prevented the decrease in the CD4/CD8 ratio in the lamina propria. CONCLUSION: Bombesin prevents the TPN-associated atrophy and dysfunction of gut-associated lymphoid tissue, supporting the concept of close neuroimmunologic interaction. PMID- 7487459 TI - Except for alanine, muscle protein catabolism is not influenced by alterations in glucose metabolism during sepsis. AB - OBJECTIVE: To assess any relationship between hyperglycemia and muscle protein catabolism associated with critical illness. DESIGN: Cohort analytic study. SETTING: Clinical research center and intensive care unit of a university hospital. PARTICIPANTS: Six healthy volunteers and five patients with severe sepsis. INTERVENTIONS: Study subjects were given infusions of 6,6,d2 glucose and 15N lysine for 6 hours. After infusion of the stable isotopes for 2 hours (basal period), dichloroacetate, which accelerates pyruvate oxidation, was given (dichloroacetate period). Leg blood flow was measured by indocyanine green dye dilution, and femoral artery and vein substrate concentrations were quantitated. MAIN OUTCOME MEASURES: The metabolic rates of glucose production, oxidation, and clearance; the whole-body protein breakdown rate; and the net efflux of amino acids from the leg were determined. RESULTS: In comparison with the healthy volunteers, septic patients had significant elevations in glucose production, oxidation, and clearance, accelerated protein catabolism, and greater net peripheral efflux of amino acids. Dichloroacetate significantly decreased glucose production and increased the percentage of glucose directed toward oxidation in both healthy volunteers and septic patients. However, this dichloroacetate induced perturbation of glucose utilization had no significant effect on whole body protein breakdown or the efflux of specific amino acids from the leg except for alanine, whose net efflux doubled (P < or = .05). CONCLUSIONS: The findings of this study demonstrate a universal acceleration in the metabolic rates of both intermediary glucose metabolism and protein/amino acid catabolism during sepsis. Except for alanine, however, there appears to be no coupling between these two physiologic responses to sepsis. PMID- 7487460 TI - Does endotoxin play a major role in inducing the depression of macrophage function during polymicrobial sepsis? AB - BACKGROUND: Endotoxin (ETX) is thought to be the primary inducer of proinflammatory mediator release associated with bacterial sepsis. Furthermore, a number of studies indicate that preexposure of animals to high doses of ETX produces macrophages (M luminal diameters) that are refractory to ex vivo stimulation with ETX. However, it is unknown if levels of ETX comparable to those typically encountered in sepsis induce a similar refractory state in M luminal diameters. DESIGN: To assess this, peritoneal M luminal diameters (PM luminal diameters) were harvested from C3H/HeN mice (ETX sensitive) at 1 hour (early) or 24 hours (late) following cecal ligation and puncture (CLP) to induce polymicrobial sepsis, sham CLP, or laparotomy followed by peritoneal implantation of a minipump delivering either saline or ETX (0.025 microgram/g of body weight, every 24 hours). Peritoneal M luminal diameter cultures were incubated with ETX, either 0 or 10 micrograms/mL, for 24 hours, and their ability to release interleukin-1, interleukin-6, and tumor necrosis factor was assessed by bioassay. RESULTS: Chronic low-dose ETX with 0 microgram of ETX media added produced early (at 1 hour) in vivo activation of PM luminal diameter interleukin-1 release, which was comparable to that seen in mice subjected to CLP. However, unlike PM luminal diameter taken from CLP mice, PM luminal diameters from mice implanted with the ETX minipump at 1 or 24 hours showed no marked decline in their ability to respond to ETX (10 micrograms). Comparable changes were seen for interleukin-6 and tumor necrosis factor release. CONCLUSIONS: Bacterial component(s) other than ETX per se induces the sustained dysfunction in PM luminal diameter capacity to produce proinflammatory cytokines during sepsis and/or peritonitis. Thus, agents directed against ETX alone may not be adequate in the treatment of polymicrobial sepsis. PMID- 7487461 TI - Tumor necrosis factor alpha gene expression in human peritoneal macrophages is suppressed by extra-abdominal trauma. AB - BACKGROUND: Trauma is believed to activate immunocytes but paradoxically also increases the risk of intraperitoneal infection. OBJECTIVE: To investigate these events by evaluating changes in the cytokine control networks of human peritoneal macrophages (PM phi) early after trauma. DESIGN: Case-control study comparing cytokine messenger RNA (mRNA) expression by PM phi from patients with extra abdominal trauma with that of both peripheral blood mononuclear cells (PBM) and PM phi obtained from healthy individuals. SETTING: Level I trauma center and basic science laboratory in a university hospital center. PATIENTS: Six patients with polytrauma (Injury Severity Score, > or = 15) with clinically negative diagnostic peritoneal lavages performed on routine indications at admission to the emergency department and six healthy age- and sex-matched individuals undergoing inguinal herniorrhaphy under local anesthesia. INTERVENTIONS: Peritoneal macrophages were isolated from diagnostic peritoneal lavages in trauma patients. Identical lavages were performed through the hernia sac in the control group. MEASUREMENTS: Cellular RNA was assayed for tumor necrosis factor alpha (TNF-alpha), interleukin-1 beta, IL-6, and IL-10 message by semiquantitative reverse-transcription polymerase chain reaction. RESULTS: Normal PM phi expressed high levels of TNF-alpha mRNA relative to PBM, but expression of the other proinflammatory cytokines was equivalent to that of PBM. Peritoneal macrophage expression of TNF-alpha mRNA was markedly (64-fold) decreased after trauma (P < .001), when PBM expression of IL-10 mRNA was increased (P = .03). CONCLUSIONS: Human PM phi constitutively show high levels of TNF-alpha message expression, and this is down-regulated by polytrauma. This might constitute a functionally "primed" state. If so, TNF-alpha down-regulation might contribute to functional PM phi suppression after systemic injury. PMID- 7487462 TI - Planned relaparotomy vs relaparotomy on demand in the treatment of intra abdominal infections. The Peritonitis Study Group of the Surgical Infection Society-Europe. AB - OBJECTIVE: To define the role of planned relaparotomy (PR) in the treatment of intraperitoneal infection, compared with that of relaparotomy on demand (RD). DESIGN: Case-control study on the basis of a prospective multicenter cohort analytic study. Statistical evaluation was done by the McNemar test for qualitative data and the Wilcoxon matched-pairs signed rank test for qualitative data. SETTING: Eighteen hospitals of different care levels in Austria, Germany, and Switzerland. PATIENTS: Thirty-eight of 42 patients with intra-abdominal infections who underwent PR were matched for APACHE II (Acute Physiology and Chronic Health Evaluation II) score, age, cause of infection, site of origin of peritonitis, and the ability of the surgeon to securely eliminate the source of infection with 38 patients taken from a cohort of 278 undergoing RD. INTERVENTIONS: Planned relaparotomy was defined as at least one relaparotomy decided on at the time of the first surgical intervention; RD, relaparotomy indicated by clinical findings. MAIN OUTCOME MEASURES: Mortality and incidence of postoperative multiple organ failure and infectious complications. RESULTS: There was no significant difference in mortality between patients treated with PR (21%) or RD (13%). Postoperative multiple organ failure as defined by a Goris score of more than 5 was more frequent in the group of patients undergoing PR (50%), compared with the group undergoing RD (24%) (P = .01), as were infectious complications (68% vs 39% [P = .01]). Infectious complications were due to more frequent suture leaks (16% vs 0% [P = .05]), recurrent intra-abdominal sepsis (16% vs 0% [P = .05]), and septecemia (45% vs 18% [P = .05]) in the PR vs the RD groups. The incidence of other complications was not different in the two groups. CONCLUSIONS: Until larger prospective studies are available, the indication for PR should be evaluated with caution. PMID- 7487463 TI - Sepsis-induced acute lung injury is attenuated by selectin blockade following the onset of sepsis. AB - OBJECTIVE: To determine the effect of infusion with a dual-binding antibody to E- and L-selectin, EL-246, in a postonset model of sepsis. DESIGN: Nonrandomized controlled study. STUDY SUBJECTS: Young Yorkshire swine. INTERVENTIONS: Three groups were studied. Controls (n = 8) received saline solution only. Untreated animals with sepsis (n = 8) received a 1-hour intravenous infusion of live Pseudomonas aeruginosa. Animals treated with EL-246 (n = 6) received the same bacterial infusion and a 2-mg/kg bolus of EL-246 at 30 minutes. OUTCOME MEASURES: Systemic and pulmonary hemodynamics, arterial blood gas determination, bronchoalveolar lavage protein and neutrophil content, neutrophil integrin and selectin expression, neutrophil oxidant burst, and organ myeloperoxidase content. RESULTS: Treatment with EL-246 significantly reduced lung injury, as indicated by improved bronchoalveolar lavage protein and neutrophil content, resulting in a significant improvement in arterial oxygenation. This reduction in lung injury was produced by a reduction in lung myeloperoxidase content. Treatment with EL 246 failed to prevent the development of pulmonary hypertension and systemic hypotension. Neutrophils from animals with sepsis exhibited significant activation and upregulation of CD18, shedding of L-selectin, and production of increased levels of oxidants compared with controls. CONCLUSION: Treatment of animals with EL-246 soon the onset of sepsis produced significant protection against acute lung injury but failed to attenuate hemodynamic derangements associated with sepsis. PMID- 7487464 TI - The possible role of a central nervous system dopaminergic mechanism in hepatic c fos protein expression following peritoneal sepsis. AB - OBJECTIVE: To investigate the hypothesis that a central dopaminergic mechanism may regulate hepatic c-fos and c-jun gene expression following peritoneal sepsis. METHODS: First, dopamine or vehicle was instilled into a stereotaxically placed intracerebral-ventricular (ICV) cannula with or without D1 (SCH 23390) or D2 (haloperidol) antagonist pretreatment in a rat model, and the effect on hepatic c fos or c-jun protein expression was investigated. Second, we investigated the effect of haloperidol and vehicle treatment following cecal ligation and puncture (CLP)-induced sepsis with respect to hepatic c-fos protein expression, c-jun protein expression, and survival. RESULTS: Intracerebral-ventricular dopamine treatment increased hepatic c-fos immunoreactive protein but had no effect on hepatic c-jun immunoreactive protein expression. Pretreatment with SCH 23390 inhibited ICV dopamine treatment-induced hepatic c-fos immunoreactive protein expression. Haloperidol pretreatment synergized with ICV dopamine treatment to overexpress hepatic c-fos protein. Haloperidol treatment significantly increased CLP-induced hepatic c-fos and c-jun protein expression and improved survival following CLP. CONCLUSIONS: Hepatic c-fos protein expression may be regulated, in part, by a central nervous system-mediated dopaminergic D1 receptor mechanism. Treatment with the D2 receptor antagonist, haloperidol, increases sepsis-induced hepatic c-fos and c-jun protein expression and improves survival following peritoneal contamination. PMID- 7487465 TI - Preemptive ganciclovir therapy in cytomegalovirus-seropositive renal transplants recipients. AB - OBJECTIVE: To evaluate the efficacy, safety, and cost of preemptive ganciclovir therapy in cytomegalovirus (CMV)-seropositive renal transplant recipients treated with antilymphocyte antibody (ALA) preparations. DESIGN AND SETTING: A prospective, randomized trial at a 650-bed tertiary medical center hospital. PATIENTS: Forty consecutive CMV-seropositive renal allograft recipients who underwent transplantation between January 1992 and January 1994 and were treated with ALA for induction immunosuppression or acute rejection therapy. MAIN OUTCOME MEASURES: The incidence and severity of CMV disease, length of hospitalization, and patient and allograft survival. INTERVENTION: Cytomegalovirus infection prophylaxis by use of intravenous ganciclovir during ALA therapy was administered to 22 patients (group 1) and the results were compared with those obtained in 18 control patients who did not receive prophylaxis for CMV disease (group 2). RESULTS: Preemptive ganciclovir therapy significantly reduced the incidence of CMV disease (P < .05) in CMV-seropositive renal transplant patients who were treated with ALA and was well tolerated. In addition, the cost of prophylactic therapy was offset by the decreased length of hospitalization observed in patients in group 1. CONCLUSION: Preemptive ganciclovir therapy provides a cost effective approach toward significantly improving the outcome of renal transplantation in CMV-seropositive patients treated with ALA. PMID- 7487466 TI - Tuberculin screening of trauma admissions at a university hospital. A preventive strategy. AB - OBJECTIVE: To determine the prevalence of tuberculosis (TB) infection in our trauma service population, namely, those patients who had positive Mantoux (purified protein derivative [PPD] [tuberculin]) tests. DESIGN: Prospective study. SETTING: Trauma service at a university hospital in Newark, NJ. PATIENTS AND METHODS: During a 9-month period, 110 admitted trauma patients underwent screening for TB. Of these patients, 20 had undergone recent PPD tests in the last 6 months, of which nine were positive. The remaining 90 patients underwent PPD and delayed-type hypersensitivity skin tests (anergy panel); five patients were unavailable for follow-up. The patients also answered questions regarding human immunodeficiency virus status, risk factors for TB infection, and clinical symptoms. RESULTS: The mean age of the study group was 34 years (age range, 13 to 69 years). Eighty-four percent of these patients were males who belonged to the high-risk socioeconomic, racial, and ethnic groups (poor, non-white). The tests for 15 patients (17%) were newly discovered to be positive for PPD. Eleven percent of the patients with negative PPD tests were anergic, thus raising the percentage of patients with positive PPD tests to 20%. The human immunodeficiency virus status was known in only 41% of the total patients. All patients with positive PPD tests had a chest x-ray film performed; of these patients, positive findings for TB were determined for one patient. Another patient had evidence of abdominal TB at laparotomy. All patients with positive PPD tests were given appointments in the pulmonary clinic for follow-up, and only three of 15 patients kept their appointments. CONCLUSIONS: Patients who were admitted to the trauma service were predominantly young males from high-risk groups with a high incidence of TB infection. Their admission to the trauma service was a unique opportunity for screening and implementing existing preventive programs. PMID- 7487467 TI - Haemophilus pneumonia is a common cause of early pulmonary dysfunction following trauma. AB - BACKGROUND: Haemophilus species are a common cause of community-acquired pneumonia; however, their significance in posttraumatic pneumonia is unclear. DESIGN: Case series. SETTING: University hospital, level I trauma center. PATIENTS: Two hundred fifty-seven consecutive patients with blunt and penetrating trauma treated for pneumonia. MAIN OUTCOME MEASURES: Length of stay in the intensive care unit, duration of ventilatory support, rate of recurrent or persistent pneumonia, and mortality. RESULTS: Ninety-six (37%) of 257 patients treated for pneumonia had a Haemophilus species isolated on sputum culture. Of these 96 patients, 49 (51%) had only Haemophilus species, while 33 (34%) had associated gram-positive organisms and 14 (15%) had gram-negative organisms. Seventeen pure cultures (29%) and seven mixed cultures (15%) (P < .05) were beta lactamase-positive trains. Compared with patients who had pneumonia caused by other bacteria, patients with Haemophilus species were younger (mean +/- SE, 35 +/- 1.7 vs 42 +/- 1.6 years; P < .05) and more severely injured (Injury Severity Score, 20.7 +/- 1.1 vs 17.5 +/- 0.9; P < .05). There were no differences in any outcome variables between the two groups. Only one (1%) of 96 patients had persistent Haemophilus species on sputum cultures after 7 days of treatment. CONCLUSIONS: Haemophilus species are a frequent cause of pneumonia following traumatic injury. This occurs primarily in the early postinjury phase and therefore should be included in the differential diagnosis of early posttraumatic pulmonary insufficiency. PMID- 7487468 TI - Clinical significance of antibiotic endotoxin-releasing properties in trauma patients. AB - OBJECTIVE: To test the hypothesis that antibiotics leading to greater endotoxin release are associated with greater mortality in septic trauma patients. DESIGN: Post hoc analysis of data from a previously conducted prospective, randomized, multicenter study designed to evaluate the efficacy of interferon gamma in preventing infection and death in trauma patients. SETTING: Nine level I trauma centers. PATIENTS: Severely injured trauma patients at high risk for sepsis. Eighty percent (N = 334) of the enrolled patients developed some manifestation of gram-negative sepsis, defined by the administration of gram-negative specific antibiotics. MAIN OUTCOME MEASURES: The in-hospital mortality rate of patients who received penicillin-binding protein 3/tumor necrosis factor (PBP3/TNF) specific antibiotics associated with the greatest degree of endotoxin release and TNF production (PBP3/TNF group, n = 78: aztreonam, ceftazidime, and cefotaxime sodium) was compared with that of patients not receiving these agents (non PBP3/TNF group, n = 256). RESULTS: Mortality in the PBP3/TNF group (17%) was higher than in the non-PBP3/TNF group (8%, P = .02). The two groups were similar in their mean (+/- SD) Injury Severity Scores (34 +/- 9), ages (31 +/- 12 years), and initial degree of bacterial contamination. CONCLUSIONS: Antibiotics that are associated with greater release of endotoxin and production of TNF are also associated with greater mortality in septic trauma patients. Decisions regarding antibiotic administration may need to consider the endotoxin-releasing properties of antibiotics in addition to their antibacterial sensitivity spectrum. Prospective studies of the effect of endotoxin-releasing properties of antibiotics on mortality are warranted. PMID- 7487470 TI - Endoscopic procedures of the upper-thoracic sympathetic chain. PMID- 7487471 TI - Transthoracic endoscopic sympathectomy in the treatment of palmar hyperhidrosis. PMID- 7487469 TI - Preoperative highly selective catheter localization of occult small-intestinal hemorrhage with methylene blue dye. PMID- 7487474 TI - Biogenic amines in silage. 3. The occurrence of six biogenic amines in farm-scale grass and maize silages. AB - Concentrations of putrescine (PUT), cadaverine (CAD), spermidine (SPD), spermine (SPM), histamine (HIS) and tyramine (TYR) in 53 grass silages and 54 maize silages were determined using an HPLC method. Concentrations of all amines excluding SPD in grass silages decreased significantly with increasing dry matter (DM) contents and decreasing acetic acid concentrations. The mean concentrations were 1310, 642, 414, 139, 120 and 33.6 mg/kg for TYR, CAD, PUT, SPM, HIS and SPD respectively in grass silages of 20-30% DM. The maximal values found were twice to five times higher than the mean values. The effect of increasing DM on the amines concentrations was not evident in maize silages. The mean concentrations, regardless of the DM content, were surprisingly high: 435, 388, 341, 71.7, 25.1 and 5.4 mg/kg for TYR, PUT, CAD, HIS, SPD and SPM. The maximal values in maize silages were three times to ten times higher than the mean values. Considering that maize silage forms the great proportion of the diets of cattle during long term winter feeding, further toxicological and physiological research should be carried out. PMID- 7487472 TI - Effect of duration of a beta-agonist treatment on growth, feed intake and carcass characteristics in finishing bulls. AB - Two experiments were conducted to study the effect of a different duration of dietary cimaterol on animal performances and carcass characteristics in finishing Belgian White-blue bulls. They received a concentrate diet ad libitum during 228 days on average, containing no cimaterol (control) or 4 ppm cimaterol during 71, 127 or 246 days (experiment 1) or during the last 76, 108 or 136 days before the end of the fattening period (experiment 2). Prior to slaughter a 6-day withdrawal period was applied. There was a significant increase in the growth rate immediately after the onset of the cimaterol feeding. A long-term cimaterol administration of 4.5 to 8 months tended to decrease daily gain towards the end of the experiment. Average daily feed intake was only slightly reduced when cimaterol was fed from the beginning of the experiment. Feed conversion was improved by cimaterol, but the effect was smaller after a long-term administration. Increased Longissimus dorsi area and hindleg width revealed muscle hypertrophy in cimaterol-treated bulls. Dressing percent and carcass meat content were increased. Optimum duration of 4 ppm dietary cimaterol varied between 2.5 and 4 months. PMID- 7487473 TI - [The influence of oral niacin doses during different dietary protein levels on indexes of rumen fermentation, blood parameters and fattening performance of young bulls]. AB - The influence of added niacin (0, 0.5 or 1 g per animal per day) and different crude protein contents (9.2 to 12.0% of DM) or the supply of various N-sources (urea, rape seed meal, soya bean meal, fish meal) were investigated on rumen fermentation, blood parameters, feed intake, weight gain and dry matter (DM) intake per weight gain in three individual feeding experiments with 156 growing bulls weighing between 175 and 300 kg per animal. Niacin supplementation did not significantly influence (P > 0.05) investigated parameters of rumen fermentation and blood. Ruminal propionate concentration increased insignificantly (from 18.9 to 19.5 moles per 100 moles on the average), inorganic P of blood serum somewhat decreased (from 2.98 to 2.82 mmol per 1) when niacin was added (P > 0.05). Protein level did not significantly influence rumen fermentation and blood parameters except an increased urea concentration in the blood of cattle fed with diets richer in protein. The DM intake of control bulls and niacin supplemented animals amounted to 6.35 and 6.46 kg per animal per day on average. Influence of niacin on DM intake varied in dependence on protein source. The daily weight gain increased from 1003 (control) to 1040 g per animal per day (+ niacin, P > 0.05). Niacin increased weight gain of bulls of urea (+ 43 g) and rape seed or soya bean meal added rations (+ 60 g per animal per day), but did not influence the weight gain in fish meal added rations. Increased weight gain resulted from ruminal and metabolic effects (about 2/3) as well as enhanced feed intake of bulls (about 1/3). Feed efficiency mostly improved. PMID- 7487475 TI - [15N-labeling of fishes using 15N isotopes in aquarium water and the effect of a different protein nutrient on the 15N elimination after the labeling period]. AB - In a preexperiment of 12 days fishes (Cyprinus carpio L.) were labelled with 15N by means of 15NH4Cl and 15N-urea resp. in the aquarium water and by feeding a protein free diet. 15NH4Cl yielded a higher atom-% 15N excess (15N') in the tissues of fishes. In the main experiment 75 fishes (Cyprinus carpio L.) were 15N labelled with 100 mg 15N'/l water from 15NH4Cl (95 atom-% 15N') in a protein free preperiod of 12 days. In the following main period the fishes received different protein sources in their diets in maintenance. A group of 20 fishes received an animal protein (fish meal) and two groups of 20 fishes each received plant proteins (soybean meal and wheat gluten resp.). The atom-% 15N' reached after the 15N-labelling period following values: digestive tract with content--7.15, liver- 5.65, gills--5.89, muscle--0.81 and chorda dorsalis--1.09 respectively. During the main period (with protein feeding) the atom-% 15N' decreased in the tissues with high protein turnover (liver and gills) on the 2nd and 4th day to 4.31 +/- 0.11 (animal protein) and 4.64 +/- 0.14 (plant proteins) in average. The corresponding values in the tissues with low protein turnover (muscle and chorda dorsalis) were 0.73 +/- 0.04 and 0.80 +/- 0.04 atom-% 15N' in average. From the measurements on the 6th, 8th and 10th day of protein feeding resulted an atom-% 15N' in average of liver and gills of 4.08 +/- 0.13 (animal protein) and 4.11 +/- 0.15 (plant proteins). In muscle and chorda dorsalis the atom-% 15N' ascended in this time upon 0.80 +/- 0.04 (animal protein) and 0.90 +/- 0.03 (plant proteins). It seems that the protein metabolism of fishes is favoured from the amino acid of plant protein in comparison to animal protein to reduce the 15N-loss of the 15N labelled body in maintenance, like the results from experiments with rats (Hernandez et al., 1981). PMID- 7487477 TI - The effect of diet composition on the development of rumen digestion in lambs. AB - The effect of composition of feed concentrate on the development of rumen digestion in suckling lambs was investigated. The experimental group of lambs was offered special concentrate diets (SCD I and SCD II), which, compared to routinely produced commercial mixtures CCD 1 and CCD 2, were completely cleared of milk protein, enriched with legumes and alfalfa meal and supplemented with cut alfalfa of the particle size 1 cm. Significantly higher concentrations of total volatile fatty acids (VFA) were recorded in experimental lambs at the age of 7, 9 and 11 weeks. The highest concentration of total VFA was observed at the age of 11 weeks (118.2 mmol.l-1 in the experimental group and 110.2 mmol.l-1 in the control group). The cellulase activity (endoglucanase E.C.3.2.1.4. and cellobiohydralase E.C 3.2.1.91.) of the rumen content of experimental lambs was significantly higher at the age of 9, 11 and 15 weeks, in comparison with the control animals, reaching the highest values at 11 weeks of age (19.32 micrograms.ml-1 in the experimental group, 15.84 micrograms.ml-1 in the control group), while the increase of number of cellulolytic bacteria in experimental lambs was insignificant. Scanning electron microscopy showed more pronounced colonization of the rumen epithelium by adherent rumen microflora in experimental lambs. Average daily weight gains over the whole investigated period reached 0.235 kg in the experimental group and 0.221 kg in the control group. PMID- 7487476 TI - Effect of selection on digestibility and carcass composition in mink. AB - In a 5-generation selection experiment, separate lines of mink (Mustela vison) were selected for litter size at 3 weeks (F-line) and body weight in September (BS-line). One unselected line served as a control (C). Nutrient digestibility was studied in a balance experiment with four male kits from each line in the last generation. Carcass composition was determined for four 4th-generation F- and BS-males each that had been killed on September 21. The feed consumption rate was higher in the BS-line animals in the balance experiment than in the F- and C animals (1687 versus 1532 and 1504 kJ/animal and day). These differences reflected the higher average live weights measured on August 25, of the BS-males (1831 g versus 1728 and 1619). CP and CHO digestibility were similar in all lines, whereas a numerically small but significant difference in apparent fat digestibility was found between the F- and BS-lines (95%) and the C-line (93%). Average body weights of animals in the carcass composition evaluation were 2607 g (BS-line) and 2023 g (F-line). Retained protein, fat, and energy were significantly affected by line. The amount of protein retained per kg metabolic weight (kg0.75) was nearly equal in the two lines, indicating that the animals' genetic capacity for protein retention was similar and probably utilized to its full extent. Retained fat and energy per kg0.75 were higher in the BS-line, indicating that the animals had been selected for fat deposition rather than for body size. Hence, selection for body size in mink should be performed by using a selection criterion with which the negative effects of increased fat deposition on reproductive performance can be avoided. PMID- 7487478 TI - Study on the energy and protein metabolism in horses. AB - The present study focused on energy and protein metabolism in pregnant and lactating mares, including the suckling and weaned growing horses, in order to determine feed availability, as also the energy and protein requirements. The authors found that the feeding diets, consisting of alfalfa hay, oats and compounds, had different availability values in terms of energy and protein, according to animal physiological conditions and age. Thus, the pregnant mares utilized the metabolizable energy (ME) and digestible crude protein (DCP) intake in average proportions of 64.5 +/- 3.2%, 54.6 +/- 3.0%, respectively, while in the lactating mares the values were 60.1 +/- 2.8%, respectively, 50.1 +/- 4.0%. However the amount of energy and protein required for maintenance was similar in these two animal physiological conditions: 531 kJ, ME/kg0.75, 529 KJ, ME/kg0.75 and 3 g DCP/kg0.75, respectively. The suckling colts had a daily average gain between 1405 and 891 g and the young horses after weaning up to 2 years of age between 773 and 447 g. The net efficiency of ME and of DCP for growth in the suckling colts was of 40.6 +/- 2.1% and 46.1 +/- 3.2%, respectively and in young horses after weaning had similar values of 40.3 +/- 4.0%, respectively 44.4 +/- 2.2%, but the maintenance requirement of energy and protein differed from 506 kJ, ME/kg0.75 and 4.51 g DCP/kg0.75 respectively to 587 kJ, ME/kg0.75 and 3.51 g DCP/kg0.75, respectively. PMID- 7487479 TI - [Determination of the protein quality of food and animal feed]. AB - The estimated value of true digestibility of food and feed proteins is in dependence from the excretion of metabolic faecal nitrogen (MFN). Results of many authors showed that a high fibre content of the diet increase the MFN-excretion and lower the true digestibility of the diet protein. The exact estimation of MFN is only possible with isotopic methods. The labelling of feed proteins with isotopic nitrogen (15N) is possible in experiments with small animals. In big animals (pigs) the utilization of recycled 15N is too high following the long transit time of non digested 15N-labelled feed protein and the parallel 15N labelling of MFN. The best method for the estimation of true digestibility of proteins is the method with 15N-labelled animals and the differentiation between nonlabelled undigested feed protein and the 15N-labelled MFN in faeces. The estimation of digestibility of essential amino acids at the end of the ileum as a measure for protein quality is not in advantage because the bacterial breakdown and synthesis of amino acids in the small intestine is unknown. The estimation of the biological value (BV) of proteins with the classical method is useful when the exact MFN was determined. The classic formula of BV [formula: see text] is only applicable in experiments with growing animals with nitrogen retention. This result of BV value is in agreement with the method only valid for growing animals with N-retention and not for animals or human beings in maintenance. The measurement of a BV of proteins with animals in maintenance is possible when the animals are labelled with 15N. The 15N-loss of the animal after the feeding of different protein sources is the smallest when the amino acid pattern of the protein is adapted to the need of amino acids for the maintenance metabolism of the animal. It was found that proteins with a high content of glutamic and aspartic acid (proteins of grains) are better proteins for the maintenance metabolism as animal proteins. Measurements with the method of the oxydation rate of essential amino acids showed that the amino acid pattern of wheat protein is in agreement with the need of amino acids for maintenance of the adult men. The evaluation of protein quality in animal nutrition for growing or lactating monogastric animals is in the present time the balance of essential amino acids in the feed protein and the need of this amino acids of the animals. PMID- 7487480 TI - [The effect of the beta-adrenergic agonist clenbuterol on the growth of skeletal muscles of rats]. AB - The effects of clenbuterol (CB) on the weights of 6 hindleg muscles as well as on structural and biochemical characteristics of extensor digitorum longus muscle (EDL) were studied on Wistar rats. Different CB-doses (5 vs. 18 mg/kg diet) were tested and rats of different sex were used. The 12 days treatment accelerated the growth of the 6 hindleg muscles by 13 to 24% without predominant response of fast twitch or slow twitch muscles. The higher muscle weights were achieved by fibre hypertrophy as demonstrated by increased fibre diameters (+13 to +16%), and not by fibre multiplication. There was no evidence for a selective fibre hypertrophy of one of the metabolic fibre types. Fibre type composition shifted to the fast glycolytic fibres (FTG +0.7 to +4.9%-units). There were no significant differences in responses between the sexes of CB-doses. Clenbuterol decreased the nucleus/cytoplasm ratio, measured as nuclei number/mm2 fibre area, between 24 and 28% and the DNA/protein ratio by 17%. RNA content and RNA/DNA ratio were enhanced by 25 and 22%, respectively; the protein/RNA ratio remained unchanged. The results suggest that clenbuterol changes fibre type composition and stimulates fibre growth in a pretranslational stage of protein synthesis without addition of nuclei by satellite cell proliferation. PMID- 7487482 TI - [Determination of true digestibility of amino acids from the small intestine of dairy cows]. AB - True digestibility of 17 amino acids was determined with 2 lactating cows, fitted with rumen fistulae and T-shaped cannulae at the proximal duodenum and terminal ileum. The calculations were based on the following regression equation: I = a + b x D + c x PFT, with I = amino acid flow at the ileum, D = amino acid flow at the duodenum, and PFT = passage of non-protein dry matter at the ileum. As the factor b means the part of undigested amino acids, 1 - b represents the true digestibility. The calculations, which comprize the results of 16 individual experiments, showed that lysine, histidine, arginine, cysteine, leucine, isoleucine and tyrosine were absorbed in the range of 85% to 90%. Lowest absorption-rates (77-80%) were found for threonine, valine, methionine, aspartic acid and serine. The overall true digestibility of amino acids was 83% without any difference between essential and not essential amino acids. PMID- 7487481 TI - Studies on the biotin requirement of broilers fed sunflower seed meal based diets. AB - A total of 560 day-old commercial broiler chicks were used in two experiments conducted to investigate the biotin requirement of broilers fed sunflower seed meal (SSM) based diets. Two basal biotin-deficient diets based on dehulled (Experiment 1) and non-dehulled (Experiment 2) SSM were each supplemented with graded levels of biotin so that the rations had 40, 80, 120, 160, 200, 240 and 280 mcg/kg feed. Therefore a total of 14 treatments were tested. Each treatment was given to duplicate floor pens with 20 chicks each for a period of 28 days. Estimation of the live weights, live weight gain, feed intake, blood glucose and free fatty acid concentrations, lipid contents and weights of liver and kidney, and liver pyruvate carboxylase activity and the records of incidence of dermal lesions, fatty liver and kidney syndrome (FLKS) mortality and leg deformities indicated that in the case of dehulled SSM, dietary biotin of of 160 mcg/kg feed was marginal while at least 200 mcg/kg appeared to be needed for optimum performance of the birds. When non-dehulled SSM was incorporated in the diet, 200 mcg biotin/kg feed was found to be the lowest dietary level needed. However, as it seems that better results could be obtained with higher levels, 240 mcg biotin/kg with non-dehulled SSM is being recommended. PMID- 7487483 TI - [The effect of the main protein source in rations of ewes and the time of blood collection on the glucose and triacylglycerol levels in blood at the beginning of lactation]. AB - The effect of the protein source of the ration (soybean meal, cottonseed cake, corn gluten and fish meal) and the time (period) of blood sample taking was examined on the content of glucose and triacylglycerols in the blood during the initial lactation period. Thirty-six ewes of the Thessaloniki crossbred type were randomly allocated to 4 groups. The ewes of each group were fed ad libitum with one of the 4 different rations, respectively. From each ewe 4 blood samples were taken in different times. The experimental design was factorial 4x4 with 9 replicates with main factors the main protein source (ration), as well as the time of blood sample taking. The protein source effect on glucose and triacylglycerol concentration in blood was not significant, while that of time of blood sample taking was significant. The interaction "ration" X "time" of sampling for the glucose and TGC concentration was not significant. PMID- 7487484 TI - Evaluation of potential sources of proteins in diets for rainbow trout (Oncorhynchus mykiss). AB - 7 diets were prepared containing as their only source of protein one of the following feeds: fish meal, casein, hydrolyzed feather meal, grieves, wheat gluten, maize gluten, soybean meal. Crystalline amino acids were supplemented except in cases of fish meal and casein to prevent specific amino acid deficiencies. Concentrations of N x 6.25 ranged between 436 and 457 g/kg dry matter (DM). Cr2O3 was added to each diet for indirect determination of digestibilities. Each diet was fed to triplicate groups of 20 rainbow trout of an average initial weight of 60 g. After 66 feeding days, all trout were killed and used for whole body analyses. Due to severely reduced intake, trout fed the casein based diet gained only about half as much weight as trout fed the fish meal based diet, though at the same fed conversion ratio of 1.0 kg gain per kg dietary DM. Gains on the other diets ranged between these two treatments with poorer feed conversion ratios. Digestibilities of crude protein of fish meal, casein, hydrolyzed feather meal, grieves, wheat gluten, maize gluten and soybean meal were: 86, 98, 67, 81, 97, 87, and 94%, respectively. The corresponding contents of digestible energy were: 21.2, 21.8, 15.4, 16.5, 19.6, 18.3, and 14.4 Mj/kg DM. Efficiency of utilization of digestible energy ranged between 43 and 54%, that of digestible crude protein between 32 and 41%. PMID- 7487485 TI - Quantitative molecular methods in virology. AB - During the past few years, significant technical effort was made to develop molecular methods for the absolute quantitation of nucleic acids in biological samples. In virology, semi-quantitative and quantitative techniques of different principle, complexity, and reliability were designed, optimized, and applied in basic and clinical researches. The principal data obtained in successful pilot applications in vivo are reported in this paper and show the real usefulness of these methods to understand more details of the natural history of viral diseases and to monitor specific anti-viral treatments in real time. Theoretical considerations and practical applications indicate that the competitive polymerase chain reaction (cPCR) and competitive reverse-transcription PCR (cRT PCR) assay systems share several advantages over other quantitative molecular methodologies, thus suggesting that these techniques are the methods of choice for the absolute quantitation of viral nucleic acids present in low amounts in biological samples. Although minor obstacles to a wide use of these quantitative methods in clinical virology still remain, further technical evolution is possible, thus making the quantitative procedures easier and apt to routine applications. PMID- 7487487 TI - Bovine respiratory syncytial virus antibodies in non-bovine species. AB - To study the role of non-bovine species in the epidemiology of bovine respiratory syncytial virus (RSV) infections, sera obtained from 9 non-bovine animal species and from humans were examined for bovine RSV specific antibodies. Sera were mainly from animals and humans which had been in contact with cattle. Forty sera of each species were tested in an RSV specific whole virus ELISA as well as in a peptide based ELISA, that was developed to measure antibodies specific for bovine RSV. Antibodies directed against RSV were detected in over 50% of sera obtained from sheep, goat, cattle and human beings, and anti-RSV activity was also found in some roe and dogs and one horse. Antibodies to bovine RSV were found in sera of all tested cattle, 11 (27.5%) goats and in some other individual animals: 3 horses, 2 roe, 1 cat and 1 dog. These results indicate that of the investigated species, besides cattle only goats might play a role in the epidemiology of bovine RSV. PMID- 7487486 TI - Serological and genomic characterisation of group A rotaviruses from lambs. AB - Four lamb rotaviruses were characterised serologically by reactions with monoclonal antibodies and genomically by hybridisation assays and sequencing. Each was found to be distinct. Three viruses belonged to the bovine genogroup and were of subgroup I. These viruses possessed serotypes G3, G6, and G10. Their corresponding P types were P1, P11, and P14 respectively. The only previous isolation of a rotavirus with VP4 of type P14 was also from lambs. The fourth isolate was G9P8, which is the first record of a G9 rotavirus from a species other than man. PMID- 7487488 TI - Characterization of two Japanese encephalitis virus strains isolated in Thailand. AB - Two strains of Japanese encephalitis (JE) virus were isolated from a pool of Culex tritaeniorhynchus captured in 1992 and another pool of Cx. vishnui captured in 1993, in Chiang Mai Area, Northern Thailand. These two strains, ThCMAr44/92 and ThCMAr67/93, could not be identified either as Nakayama or JaGAr01 subtype by the hemagglutination-inhibition (HI) and the neutralization (N) tests using immune sera raised against these standard JE virus strains. Reverse transcription polymerase chain reaction showed the presence of JE-specific conserved sequences in these strains. Sequencing of 240 nucleotides in their PrM gene region identified that these two strains belong to the genotype 1 of JE virus. Nucleotide and encoded amino acid sequences of their envelope glycoprotein gene revealed 98.8 and 99.8% identity, respectively. These two strains shared 77.8 to 87.7% homology in the nucleotide sequence and 90.0 to 98.8% homology in the amino acid sequence with other reported JE strains. Five strain-specific amino acid changes were noted in ThCMAr44/92 strain, while one in ThCMAr67/93. In addition, four common amino acid changes were found in both strains. Thus, the findings indicated that these two strains were structurally different from each other as well as different from all the reported strains which was in agreement with the serological tests by hemagglutination-inhibition and neutralization. PMID- 7487489 TI - The complete nucleotide sequence and genome organization of odontoglossum ringspot tobamovirus RNA. AB - The complete nucleotide sequence of the genomic RNA of odontoglossum ringspot tobamovirus (ORSV) was determined. The RNA genome of ORSV is 6618 nucleotides long and contains five open reading frames (ORFs 1 to 5) coding for proteins of M(r) 126 K, 181 K, 34 K, 18 K and 52 K, respectively. This is the longest RNA of the known viruses of the Tobamovirus genus. The sequences of the ORSV RNA encoded proteins exhibit high homology to the proteins of the members of the Tobamovirus genus. The genomic organization and sequence analysis showed that ORSV is more closely related to tobacco mild green mosaic virus (TMGMV), pepper mild mottle virus (PMMV), tomato mosaic virus (ToMV) and TMV than to cucumber green mottle mosaic virus (CGMMV) and sunn-hemp mosaic virus (SHMV). PMID- 7487490 TI - The complete nucleotide sequence of the Popp (1967) strain of Marburg virus: a comparison with the Musoke (1980) strain. AB - The nucleotide sequence of genomic RNA of Marburg virus strain Popp was determined. Strain Popp was isolated in 1967 during the first filoviral outbreak. The virus was purified from blood of infected guinea pigs in which it had been maintained. The length of the determined sequence was 19112 nucleotides. Amino acid sequences of seven known virion proteins were deduced. Nucleotide and amino acid sequences were compared with those of strain Musoke of Marburg virus isolated in 1980 in Kenya and purified from Vero cells. Homology between nucleotide sequences of two strains was 93.9%. Comparisons revealed conserved and variable regions of the nucleotide and amino acid sequences. The GP, the envelope protein of the virion, was found to be the most variable protein. The greatest differences in the protein were located in the supposedly external part of the molecule. Amino acid substitutions in the L protein, the main component of viral RNA-dependent RNA polymerase, were also distributed extremely non-randomly. It was shown that the non-coding regions of the genome were more variable than the coding ones; 37.6% of nucleotide differences corresponded to the former. 72.6% of nucleotide substitutions located in the coding regions were found to be at the third codon position. PMID- 7487491 TI - Membrane related effects in endothelial cells induced by human cytomegalovirus. AB - Previously, we have reported on the increase in procoagulant activity of human umbilical vein endothelial cells (HUVEC) after infection with human cytomegalovirus (HCMV). When using microvascular endothelial cells from foreskin (MVEC), we also observe a significant increase in membrane perturbation and a concomittant increase in procoagulant activity. This effect is both observed with a laboratory HCMV strain (AD169) with low pathogenicity for endothelium and a HUVEC adapted strain (VHL-E) that readily infects endothelial cells. We compared the membrane perturbation of two types of endothelial cells, HUVEC and MVEC with human embryonal fibroblasts (HEF), being fully permissive for both strains. A membrane effect was only found in endothelial cells. Our results suggest that HCMV induces in MVEC more merocyanine-540 incorporation in the membrane as in HUVEC. The increase in the procoagulant activity induced by HCMV was more pronounced in MVEC than in HUVEC. Inactivated virus, as well as virus pre incubated with heparin was unable to evoke membrane perturbation. It therefore appears that HCMV induces a rapid membrane response in vascular endothelium and that physical interaction of the virion and the endothelial cell is required to elicit this response. PMID- 7487492 TI - Interaction of reconstituted Sendai viral envelopes with sperm cells: reconstituted Sendai virus envelope-induced fusion-mediated introduction of foreign material into bull sperm cells. AB - Reconstituted Sendai virus envelopes (RSVE), i.e. membrane vesicles bearing the viral envelope glycoproteins and phospholipids, are able to fuse with bull sperm cells. This was inferred from the increase in the degree of fluorescence dequenching (DQ) obtained following incubation of fluorescently labeled (R18 labeled) RSVE with bull sperm cells and from electron microscopy studies of RSVE sperm interaction. Only a low degree of DQ was observed, under the same conditions, with non-fusogenic fluorescently labeled RSVE. This, and electron microscopy results, show that binding and membrane fusion events occur between RSVE and sperm cells. In addition, DQ was observed following incubation of RSVE that had been pre-loaded with the self-quenched fluorochrome Calcein, with bull sperm cells, indicating fusion-mediated injection of the dye from the RSVE space into the sperm cells. PMID- 7487493 TI - Selective transmission of hepatitis C virus in vivo and in vitro. AB - A human plasma containing quasi-species of hepatitis C virus (HCV) was inoculated to a chimpanzee and to human lymphocytic cell lines, HPB-Ma clone 10-2, AD HPB, and Daudi, which support replication of HCV. Among six different hypervariable region (HVR) sequences detected in the inoculum, the same two were recovered both in vivo and in vitro. PMID- 7487494 TI - Induction of protective antibody responses against pseudorabies virus by intranasal vaccination with glycoprotein B in mice. AB - Intranasal vaccination of mice with glycoprotein B (gB) of pseudorabies virus (PRV) induced specific IgA and IgG antibody responses in the secretion of the respiratory tract, resulting in protection of the animals against intranasal challenge with a lethal dose of virulent PRV. The immune response was enhanced by the use of cholera toxin B subunit as an adjuvant. The present results indicate that local vaccination with gB is a promising strategy to confer protective immunity on animals against PRV infection by inducing secretory antibodies on their mucosal surfaces where the primary replication of the virus occurs. PMID- 7487495 TI - The US3 protein kinase of herpes simplex virus type 2 is associated with phosphorylation of the UL12 alkaline nuclease in vitro. AB - Herpes simplex virus type 2 (HSV-2) gene US3 encodes a serine-threonine protein kinase. We previously described the isolation of a US3-inactivated mutant which is able to replicate in Vero cells but not in murine macrophages. To learn more about the biological role of the US3 protein kinase, we have sought to identify the target proteins of the enzyme. Studies of in vitro phosphorylation with extracts of infected cells demonstrate that the US3 protein kinase is involved in phosphorylation of the UL12 alkaline nuclease in vitro, suggesting that the nuclease is a possible target of the protein kinase. PMID- 7487496 TI - In vitro synthesis of foot-and-mouth disease virus specific antibodies by porcine leukocytes. AB - We have characterized the in vitro secondary antibody response to FMDV of peripheral blood mononuclear cells (PBMC) from immunized pigs. The results obtained indicated that primed swine leukocytes can support an in vitro T-B cell cooperation which is functional and leads to the production of viral specific antibodies. The response was shown to be independent of viral replication, being induced by both infective and inactivated virus as well as by recombinant polypeptides VP1 and VP3. In all cases, concentration of PBMC supernatants allowed the detection of viral-specific IgG antibodies by ELISA. Significant titers of foot-and-mouth disease virus (FMDV) specific neutralizing antibodies were detected in concentrated supernatants after stimulation with either infective or inactivated whole virus, whereas no neutralizing activity was found in supernatants from PBMC responding to individual capsid polypeptides. The titers of IgG1 and IgG2 were similar for PBMC incubated with viruses, while IgG2 predominated when VP1 or VP3 were used as stimulators. In addition, significant titers of IFN-gamma were detected in supernatants of PBMC stimulated with infectious or chemically inactivated FMDV. PMID- 7487497 TI - The nucleotide sequence of asinine herpesvirus 3 glycoprotein G indicates that the donkey virus is closely related to equine herpesvirus 1. AB - The nucleotide sequence of the glycoprotein G (gG) homologue of asinine herpesvirus 3 (AHV3), a respiratory alphaherpesvirus of donkeys, was determined. The AHV3 gG gene consists of 1233 base pairs (bp) and codes for a predicted protein of 411 amino acids. This is identical in size to the equine herpesvirus 1 (EHV1) gG gene and 6 amino acids longer than the equine herpesvirus 4 (EHV4) gG gene. The predicted amino acid sequence of AHV3 gG has characteristics of a class 1 membrane protein. The amino acid sequence of AHV3 gG shows 92% and 60% identity to EHV1 gG and EHV4 gG respectively. Two regions within the gG amino acid sequences of EHV1 and EHV4 were previously defined, an N-terminal constant region and an immunodominant highly variable region located toward the C-terminus. In the corresponding constant region of AHV3 gG there was 96% and 75% amino acid identity with EHV1 and EHV4 gGs respectively. In the variable region, there was 73% and 24% identity respectively. Phylogenetic analyses using the gG nucleotide sequences indicated that AHV3 is much closer in evolutionary distance to EHV1 than either virus is to EHV4. These findings provide additional support for the view that AHV3, or another closely related virus, may be the progenitor of EHV1 and has adapted to horses in relatively recent times. PMID- 7487498 TI - Cloning and restriction endonuclease mapping of herpes simplex virus type-1 strains H129 and +GC. AB - EcoRI fragments of herpes simplex virus I (HSV-1) strains H129 and +GC were cloned and the EcoRI and BglII restriction enzyme sites were mapped. Comparison of these enzyme sites with the sequence of HSV-1 strain 17syn+ demonstrated that all EcoRI sites were identical. For H129, the BglII sites were also found to match strain 17syn+ BglII sites. With one exception, the BglII sites in strain +GC also aligned with the strain 17syn+ sequence. The one exception was a missing BglII site from strain +GC located between bases 25,149 and 25,154 in the EcoRI D fragment within the viral deoxyribonuclease gene (UL12). The BglII site represents the first difference to be mapped within HSV-1 strains H129 and +GC which have unique pathobiological properties in animal models of acute and reactivated infections. PMID- 7487499 TI - A conserved, precise RNA encapsidation pattern in Tobamovirus particles. AB - The bidirectional RNA encapsidation pathway in nine sequenced Type 1 Tobamovirus genomes will result in RNA-coat protein assembly, up to and including the first transcribed G, adjacent to the 5'-cap structure (m7 Gppp). This precision is highly conserved, despite wide interstrain variations in the absolute position of the phase-determining core of the origin-of-assembly sequence (Gxx)n and in overall genome length (6311-6507 nts). A Type 2 Tobamovirus genome did not comply with this pattern. All genomes had a statistically significant bias for G at every third (or 3n) position, resulting in a preponderance of GNN codons and hence a high Val, Ala, Gly, Asp, Glu content, at least in the large (126/183 kDa) and amino-coterminal replicase protein genes. Contrary to predictions from the X ray fibre diffraction structure of tobacco mosaic virus (TMV, U1 strain), only one (pepper mild mottle virus) of the nine Type 1 Tobamoviruses positioned the preferred G-repeat in the most favourable (5') position of the trinucleotide binding site on each coat protein (CP) subunit. In all but one of the eight remaining Type 1 Tobamovirus genomes, G would predominate in the CP 3'-site. The significance of these observations for TMV particle assembly, disassembly and host cell interactions are discussed. PMID- 7487501 TI - [Management of newborn infants with heart disease. The persistent challenge]. PMID- 7487500 TI - The National Institute for Medical Research, Mill Hill. Personal recollections from 1956/57. PMID- 7487502 TI - [Alternative pathways of angiotensin I. The glimpse of new possibilities in the treatment of hypertension and heart failure]. PMID- 7487503 TI - [Management of children with heart murmurs. Diagnostic importance of noninvasive complementary tests]. AB - PURPOSE: To determine the benefits of noninvasive tests in the characterization of heart disease in children referred due to heart murmur. METHODS: Two hundred and thirty three consecutive children were fully examined with the aid of the electrocardiograma (ECG), chest X-ray and echocardiogram (ECHO). The patients were divided in 2 groups according to the initial diagnosis of innocent murmur (n = 23) and pathological murmur (n = 210). A comparison was made between the initial and final diagnosis after the noninvasive tests. Statistical analysis was employed through the qui-square test. RESULTS: The initial diagnosis did not change after ECG. The chest X-ray induced the diagnosis of idiopathic dilatation of the main pulmonary artery in one case and of cardiomyopathy in 8 cases, confirmed in only one. After ECHO, the initial diagnosis of innocent murmur was maintained in 70% of the cases, while 30% had some form of heart disease. In 80% of the pathologic murmur group, the diagnosis did not change after ECHO, while 20% were considered as having an innocent murmur. In the 2 groups together, the concordance index between initial and final diagnosis after ECHO reached 90%. Statistically, a non-significant difference was found between the 2 groups. CONCLUSION: Careful clinical examination is mandatory in the initial evaluation of children with heart murmur. ECG and chest X-ray does not change the initial clinical diagnosis. After ECHO, the diagnosis changing was low (10%), as long as clinical mistakes are eliminated. The initial diagnosis of innocent murmur excludes the necessity of noninvasive tests with a small possibility of error. EKG, chest X-ray and ECHO should be reserved to cases with obvious or persistent suspicious heart disease after clinical examination. PMID- 7487504 TI - [Early physical conditioning of patients surgically treated for myocardial revascularization. Influence on the functional capacity]. AB - PURPOSE: To evaluate the early physical conditioning (PC) effect [initiated 15 days after the myocardial revascularization surgery (MRS)] in the functional capacity. METHODS: Twenty-two male patients (mean-age of 52y-o), divided in two groups (A and B, 11 patients each), were studied. Group A started PC 3 months after MRS (phase III), with a training intensity of about 70% of the maximum heart rate reserve, during three months, three sessions per week with one hour duration. Group B started PC 15 days after the MRS (fase II), with a training intensity up to five metabolic unities, three times a week, 1 hour and 15 duration. The post-MRS period of three months was designated as the time I (beginning of fase III) and six months as time 2. Stress test, Kraus-Weber flexibility test, coxo-femural flexibility test (flexion, elevation and abduction) and scapule-umeral flexibility test (flexion, extension and abduction) were applied. RESULTS: The maximum oxygen uptake and the total work increased significantly from time 1 to time 2 in both groups, but there was no significant difference between the two groups, either in time 1 or 2. Flexibility in the Kraus-Weber test and in the scapule-umeral flexibility test when in flexion, were significantly greater in group B than A, in both times (1 and 2). Other flexibility variables studied show no significant difference, neither between the studied groups nor in the times 1 and 2 of evaluation. CONCLUSION: Early PC after MRS increased the value of two variables that measure flexibility. However, it has not increased other flexibilities variables and the aerobic capacity. PMID- 7487505 TI - [Heart surgery with extracorporeal circulation in pregnant women. Analysis of materno-fetal outcome]. AB - PURPOSE: To analyze maternal and fetal outcome in pregnant undergone to cardiac surgery. METHODS: We studied the evolution of 30 pregnant women submitted to cardiac surgery at the Hospital Sao Paulo, between Jan/81 and Dec/92 and, further, attended at this Hospital till the parturition. The following variables were analyzed: cardiopulmonary bypass, time of the procedure and time of the anoxia, patient temperature, surgical complications, and neonatal, maternal and fetal outcomes. RESULTS: All patients had rheumatic heart disease and, in 17, mitral stenosis was the main anatomic abnormality. Mitral commissurotomy was performed in 24 patients, double comissurotomy (mitral and aortic) in 1 patient and valve replacement was performed in 5. Cardiopulmonary bypass was utilized in all procedure; occurrence of surgical complications (p < 0.001) and the prolonged surgical time (p = 0.009) were related to the fetal mortality. There was 4 (13.3%) maternal deaths and 10 (33.3%) fetal deaths related to the surgery. CONCLUSION: The indication of cardiac surgery in pregnant women is heart failure, refractory to conventional therapy; cardiopulmonary bypass is associated with high fetal mortality. PMID- 7487507 TI - [Myocardial revascularization in left coronary trunk lesion in patients over 65 years old]. AB - PURPOSE: To study the short and long-term follow-up of patients with left main coronary artery disease (LMCAD) and age over 65 years, by comparing the results with patients under 65 years-old. METHODS: Twenty-two patients with LMCAD and mean age of 69 +/- 3.5 years (group I) were underwent isolated coronary artery bypass grafting (CABG) and compared to 31 patients with LMCAD, mean age of 54 +/- 7 years (group II), who also underwent isolated CABG. The life-table Kaplan-Meyer method was used to estimate the post-operative survival. The chi-square and Student "t" test were used when necessary. RESULTS: Despite higher operative mortality in group I (9.1% x 3.2%), the difference was statistically not significant. The operative morbidity was similar in both groups. Actuarial survival at 4 years was 85% in group I and 95% in group II. Actuarial survival free of cardiac events was 69% in group II and 75% in group II. CONCLUSION: The CABG is well tolerated and had low morbidity and acceptable mortality in old patients with LMCAD. The long-term survival in these patients was very similar to the younger patients. PMID- 7487506 TI - [Mortality trends due to myocardial ischemia in capital cities of the metropolitan areas of Brazil, 1979-89]. AB - PURPOSE: To study the trends of mortality rates from cardiovascular diseases (CVD) and coronary heart disease (CHD) in eight capitals of Brazil. METHODS: Death rates for these cities were studied in men (M) and female (F), standardized using the population of Sao Paulo, SP, 1980. Rates were calculated for CVD (ICD 9: 390-459) and CHD (ICD-9:410-414). The cities were Belem, Recife, Salvador, Belo Horizonte, Rio de Janeiro, Sao Paulo, Curitiba and Porto Alegre. Using simple linear regression, we obtained the annual difference in rates with a confidence interval of 95%. RESULTS: For CVD a decline was detected of -2.6% (M) and -2.69% (F) in Belem; in Salvador, -1.53% (M) and -1.70% (F) and in Sao Paulo, -1.27% (M) and -1% (F) and for F in Belo Horizonte (-0.82%) and in Curitiba ( 1.52%). Recife, Porto Alegre and Curitiba (M) showed stability during the period. There was a rise of the rates in Rio de Janeiro +1.37% (M) and +0.84% (F) and for M in Belo Hozironte (1%). The rates for CHD showed a decline in Belem-2.64% (M) and -2.74% (F); Sao Paulo, -1.4% (M) and -1.06% (F) and for F in Belo Horizonte. A stable trend was detected in Salvador, Curitiba, Porto Alegre and for M in Belo Horizonte. There was a rise in the rates of Recife +3.5% (M) and +4.4% (F) and in Rio de Janeiro +2.34% (M) and +2.53% (F). CONCLUSION: The magnitude of death rates and of the time trends showed in this paper reveal important regional differences. Questions related to the different assignment of diagnosis in the chapter of CVD could explain some of the observed trends. However, the rise of mortality rates in Rio de Janeiro could hardly be explained by classificatory matters. Tobacco, diet and medical care could explain the rise of mortality from CVD and CHD in Rio de Janeiro. PMID- 7487508 TI - [Significance of the nuclear magnetic resonance in the detection of coronary artery patency after thrombolysis]. AB - PURPOSE: To determine the value of magnetic resonance imaging (MRI) in the noninvasive detection of infarct related coronary artery patency after thrombolysis. METHODS: We studied 26 patients with acute myocardial infarction submitted to thrombolysis underwent MRI studies before and after 0.1mmol/kg gadolinium-DTPA injection within the first 48 h of MI. Signal intensity was assessed by circumferential profile analysis techniques. RESULTS: The average ratio of signal intensity of infarcted tissue over normal myocardium (I/N) was significantly higher in patients with patent arteries (1.3 +/- 0.13 vs 1.12 +/- 0.07, p < 0.02). Compared to coronariography MRI, sensitivity of 81% and specificity of 100% for the diagnosis of coronary patency. CONCLUSION: Gadolinium infusion increased infarcted and normal myocardium differentiation. The study of gadolinium kinetics at MRI is a promising technique for noninvasive diagnosis of coronary patency. PMID- 7487509 TI - [Influence of mannitol added to the nutrient solution on the mechanical performance and on the degree of myocardial edema of isolated hearts of rats]. AB - PURPOSE: To analyse the influence of mannitol added to Krebs-Henseleit (KH) solution on the myocardium edema and myocardial function. METHODS: Isolated rat heart under isovolumetric contractions studied according to Langendorff's technique were perfused with KH solution at constant flow during 90 min. The coronary perfusion pressure, diastolic and systolic pressures were recorded at every 15 min. At the end of the experiment, myocardium water content was measured in hearts perfused with KH solution (group I, n = 9) and in hearts perfused with KH solution plus 8mM mannitol (group II, n = 8). These results were compared to non-perfused control heart (n = 9). RESULTS: Myocardial water content was statistically higher in group I (80.8 +/- 1.3%) compared to group II (78.1 +/- 0.7%) and control group (75.5 +/- 0.5%). Systolic arterial pressure was statistically higher in group I (86.2 +/- 11.5mmHg) compared to group II (72.7 +/ 21.1mmHg). There was no difference in the diastolic pressure between the two groups. Coronary perfusion pressure (Pp) increased progressively during the experiment in both groups. However, Pp was lower in group II than in group I. CONCLUSION: Mannitol added to KH solution significantly attenuates the myocardium edema in the isolated perfused rat heart. PMID- 7487511 TI - [Case 2/95 (Faculdade de Ciencias Medicas da Unicamp)]. PMID- 7487510 TI - [Pulmonary atresia with intact ventricular septum. Valvar perforation by radiofrequency]. AB - Six days old neonate with pulmonary atresia and intact ventricular septum is reported in which a communication was established between the right ventricle and the pulmonary artery. The membraneous atresia was perforated with a special radiofrequency catheter 2F connected to a Hat 200S radiofrequency generator (Osypka). The energy necessary to perforation was 15W. After perforation of the atresia, balloon dilation was performed (balloon diameter: 2, 3, 4mm of coronary angioplasty and 5, 8mm Penta Medi-tech). The valve was completely opened giving the continuity between right ventricle and pulmonary artery. PMID- 7487512 TI - [Effects of aerobic physical conditioning on blood pressure control]. PMID- 7487513 TI - [Role of ambulatory blood pressure monitoring in the evaluation of the therapeutic efficacy of antihypertensive drugs. Trough/peak relation]. PMID- 7487514 TI - [Chagas disease. Importance of the parasite in the pathogenesis of the cardiac chronic disease]. PMID- 7487516 TI - [Child with heart murmur. Importance of the noninvasive evaluation]. PMID- 7487515 TI - [Unified hypothesis about the pathogenesis of chronic Chagas cardiopathy. Therapeutic implications]. PMID- 7487517 TI - [Heart surgery with extracorporeal circulation in pregnant women]. PMID- 7487518 TI - [Sudden death. Mechanisms and prevention]. PMID- 7487519 TI - [Thrombolysis (Consensus about post-myocardial infarction treatment)]. PMID- 7487520 TI - [Management of the patient with infarction after hospital discharge (Consensus about post-myocardial infarction treatment)]. PMID- 7487521 TI - [Rehabilitation after acute myocardial infarction (Consensus about post myocardial infarction treatment)]. PMID- 7487522 TI - [Serum enzymes and electromyography in neuromuscular diseases: comparative study of 817 cases]. AB - This study reports the relationship of the serum enzymes (creatinekinase 817 cases, lactic dehydrogenase 784 cases, aldolase 718 cases, aspartate aminotransferase 767 cases and alanine aminotransferase 760 cases) and electromyography (EMG) of 588 cases (20 normal, 299 with myopathic pattern, 209 with denervation and 69 with neuromyopathic pattern) in several neuromuscular disorders. The relationships were studied using descriptive statistic and chi square tests. It was found a statistical significance with the increased serum enzyme level with the myopathic EMG pattern and an inverse relationship with the denervation EMG. This relation was more important with the creatinekinase, following aldolase and lactic dehydrogenase. The EMG denervation pattern did not have any relation with serum enzyme levels. PMID- 7487523 TI - [Myasthenia gravis: results of thymectomy in 52 patients]. AB - From 1971 to 1993, 52 myasthenics without thymomas (7 men and 45 women) were thymectomized. The results were categorized in remissions and non-remissions. The remission rate was 48%. In the remission group there were 5 men and 20 women and in the non-remission group 2 men and 25 women. The follow-up time was 5.5 years in both groups. There were 16 patients in Osserman category II A, 11 achieving remission; 36 patients were in categories IIB and III, and 14 achieved remission. The duration of disease was 1.8 years and 4.3 years in the remission and non remission groups, respectively. All patients with remission had less than four years of disease but among the 43 patients with less than four years of symptoms, 42% did not achieve remission. Our data reinforce the need for early thymectomy in myasthenia gravis. PMID- 7487524 TI - [Multiple sclerosis: clinical analysis and development of 214 cases]. AB - The authors report on a series of 214 cases of defined multiple sclerosis according to Poser et al. criteria. These cases were retrospectively selected by medical reports analysed from Santa Casa Medical School (Sao Paulo) and the private practice, from 1980 to 1993. The data were analysed as for sex, race, onset age of symptoms, onset symptoms, and evolutive symptoms. The results were compared to those found in other Brazilian series and in foreign series. This comparative analysis allows to verify similarities and differences among the several series, and the authors emphasize the necessity of multicentric studies in Brazil to assert with more details the multiple sclerosis profile in our country. PMID- 7487525 TI - [Epilepsy with centro-temporal spikes and parietal spikes: comparative study]. AB - There are controversies about the existence of a benign parietal epilepsy distinct from the benign partial epilepsy with centro-temporal spikes. We studied 164 children with no neurological or neuroradiological evidence of brain damage and with epilepsy and spikes restricted to the centro-temporal (CTS) or to the parietal regions (PS). The subject's age, age at onset and type of seizures and the presence of spikes evoked by consecutive taps applied to both hands and feet were compared between 111 patients with CTS and 53 patients with PS. Age of patients and age at onset of seizures predominated until 6 years in the PS group and after 6 years in the CTS group. The occurrence of oropharyngeal or facial motor seizures was statistically more frequent among the patients with CTS (44.1%) as opposed to PS group (16.9%). In 39.5% of the children with PS and only in 3.5% of those with CTS, the EEG showed high voltage potentials, similar to the habitual spikes in clinical EEG, evoked by the stimulation of one or both feet or hands. Our findings suggest that in neurologically normal children with epilepsy, the group with PS differs from those with CTS in the age, age at onset and type of seizures and EEG reactivity to percussion of hands and feet. PMID- 7487526 TI - HTLV-I associated myelopathy in patients from Brazil and Iran: neurological manifestations and cerebrospinal fluid findings. AB - We analysed sera and cerebrospinal fluid (CSF) of 20 cases of human T-cell lymphotropic virus type (HTLV-I) associated myelopathy/tropical spastic paraparesis (HAM/TSP) from Brazil and Iran and as controls, 16 Brazilian HTLVI seronegative individuals afflicted with other neurological diseases. It was observed in the HAM/TSP patients that: 1) all had an inflammatory reaction within the central nervous system (CNS); 2) 95% (19/20) showed oligoclonal bands reflecting intrathecal IgG synthesis; 3) 85% (17/20) presented a local synthesis of HTLVI antibodies; 4) 35% (7/20) had a measurable immunoglobulin (Ig) synthesis within the CNS. The CSF parameters of the HAM/TSP were compared with the clinical data (age at onset, duration of disease and disability level). Our data prove that CSF analysis is important for the diagnosis of HAM/TSP. There is no association between the severity of the disease and CSF findings. PMID- 7487527 TI - Cerebrospinal fluid cytomorphologic findings in 41 intracranial tumors: a retrospective review. AB - The main objective of this retrospective review of clinical and cerebrospinal fluid (CSF) data from 41 patients with intracranial tumors diagnosed between 1975 and 1989, is to report the role that the finding of neoplastic cells in CSF plays, specially when cerebral CT-scanning and MRI were not currently done. Another objective is to study the CSF proteic abnormalities in cerebral tumors. CSF cell count, cytomorphologic pictures obtained after sedimentation and protein findings are described. Tumor cells were seen in 12 cases (29%): medulloblastomas -6, meningeal carcinomatosis--3, multiforme glioblastoma--1, ependymoma--1, cerebral metastasis--1; in two cases it was an unexpected finding. We noticed that tumoral localization next to the ventricles favoured cell exfoliation. Although pleocytosis was rare and uncorrelated with the presence of neoplastic cells, pathological cytomorphologic pictures appeared in most of the cases including all "positive" ones. Our results stress that the appearance of neoplastic cells in CSF remains helpful specially when it is an unexpected finding. PMID- 7487528 TI - [Occurrence of cysticercosis in autopsies performed in Uberlandia, Minas Gerais, Brazil]. AB - 3937 autopsies were performed between 1971 and 1993 in the Servico de Anatomia Patologica of the Hospital de Clinicas of the Fundacao de Assistencia, Estudo e Pesquisa de Uberlandia, Universidade Federal de Uberlandia, in Minas Gerais, Brazil. At this Service of Pathology are realized all the autopsies of the municipal district of Uberlandia. The analysis of 2862 concluded autopsy reports, of death above the age of one year, disclosed 39 cases (1.4%) of cysticercosis. The age range was 16 to 83 years and 66.6% were males; 82.1% of the patients were from Minas Gerais State, 15.4% were from Goias State, and in one case (2.5%) the origin was not registered. From these 39 cases, 35 (89.7%) showed central nervous system involvement, isolated or in association to other clinical forms of the disease; in 9 occurred the isolated or associated cardiac form; in 4 the muscular form, isolated or associated, was found; 4 presented the isolated or associated visceral form. In only 7 (17.9%) cases, the cysticercosis was assumed to be the direct cause of the death. PMID- 7487529 TI - [Myometric evaluation of patients with Duchenne muscular dystrophy]. AB - An account of the authors' experience in strength measurement using a hand-held dynamometer in 16 patients with Duchenne muscular dystrophy (DMD) is given. A rapid decrease of knee extension strength was observed, between 6 and 8 years of age, analysing among patients of different ages. At the same time loss of the ability to walk has occurred. An unexplainable increase in strength was observed in two patients examined in a six month interval. A short review of the literature is given and the conclusion of the importance on the wider use of the instrument. PMID- 7487531 TI - [Apomorphine: an alternative in the control of motor fluctuations in Parkinson's disease]. AB - Levodopa-induced motor fluctuations (MF) is a disabling complication of Parkinson's disease (PD) and is usually refractory to conventional treatment. Apomorphine, a dopamine agonist with affinity for both D1 and D2 receptors, has been emerged as an useful alternative in the management of MF of PD. The frequency of nausea and vomiting prevented its use in the past, but the simultaneous administration of domperidone has proved to be able to control these side effects. Although apomorphine has been successfully used to control levodopa induced MF in other countries, it has not been considered in the management of PD in Brazil. We report here our initial experience with subcutaneous injections of apomorphine combined to oral domperidone. We administered apomorphine in doses ranging from 1.5 to 3 mg in four PD patients with MF of our outpatient clinic. All the doses administered switched the "off" state to a motor response qualitatively similar to what is seen in the "on" phase induced by levodopa, including the occurrence of dyskinesia. The latency to turn "on" after apomorphine ranged from 7 to 30 minutes and the duration of the response ranged from 60 to 85 minutes. We observed yawning in all four patients, labial paresthesia in one patient and an inspecific unpleasant sensation in another patient. These side effects were not significant in our four patients. Our data show that the use of apomorphine adds a reliable and effective strategy in the management of MF of PD patients. PMID- 7487530 TI - [Smoking and mortality from cerebrovascular disorders in Brazil: comparative study of capital cities of metropolitan regions, 1988]. AB - AIM: The role of smoking as risk factor for stroke is controversial. An ecological design study was performed to test the association between stroke mortality and smoking in Brazil. Lung cancer mortality was used as a surrogate for smoking habit. METHODS: The mortality rate for stroke (ICD-9:430-438) and lung cancer (ICD-9: 162) were determined in the following metropolitan areas: Belem, Recife, Salvador, Belo Horizonte, Rio de Janeiro, Sao Paulo, Curitiba and Porto Alegre for males and females between 30 years-old and 69 years-old. ANOVA test was used to compare both age-adjusted mortality rates. RESULTS: A negative association by ANOVA was determined between age-adjusted rates and age-specific rates, except among males with 40-49 years-old. CONCLUSION: These results may be on account of three causes: the different smoking role as a risk factor in the distinct stroke subtypes analyzed; the competitive risk between coronary heart disease and stroke; and the high prevalence of arterial hypertension in the Brazilian metropolitan areas. PMID- 7487533 TI - Atypical EEG pattern in children with absence seizures. AB - We studied four children with diagnosis of absence seizures (generalized primary epilepsy), and with a generalized delta activity on the EEG during clinical attacks provoked by hyperventilation. The lack of ictal generalized spike-and wave discharges with a frequency of 3 Hz in our patients, makes this an atypical pattern. All children had complete control of their seizures and disappearance of the EEG changes with valproate. We concluded that generalized delta activity observed on EEG during the hyperventilation in children should not always be considered as a normal finding for age, since it could be an ictal event of an absence seizure. PMID- 7487534 TI - Atypical pattern related to 14 Hz positive spikes. AB - We studied two children with a history of headache and a normal physical and neurological examination whose EEG showed an electroencephalographic pattern recently published, the N-shape potential associated with the 14 Hz positive spikes. This graphoelement was observed only during the asleep state. PMID- 7487535 TI - Subclinical rhythmic electrographic discharge in adults: an atypical evolution. AB - The subclinical rhythmic electrographic discharge in adults (SREDA) has been rarely reported. We describe the case of a 71 year-old man with transient ischemic attack, whose EEG showed an atypical evolution of this uncommon pattern. The computed tomography scan and cerebrospinal fluid were normal. We are not aware of any other report in the literature with this atypical evolution of SREDA. PMID- 7487536 TI - Dipole reversal: an ictal feature in a patient with benign partial epilepsy of childhood with centro-temporal spike. AB - We describe the case of a 15-year-old boy who had the diagnosis of benign partial epilepsy of childhood with centro-temporal spike. During the EEG a subclinical electrographic seizure was recorded. The discharges were clearly electropositive in T4 with positive phase reversal between derivations F8-T4 and T4-T6. The whole episode lasted less than one minute (45 sec). The interictal right medio-temporal spikes reemerged after 60 sec and were electronegative in the same location after the end of the electrographic seizures. The mechanisms underlying this uncommon pattern on EEG is not well established. PMID- 7487532 TI - Epilepsy with continuous spike-waves during slow wave sleep: a clinical and electroencephalographic study. AB - We report four children with epilepsy with "continuous spike-waves during slow wave sleep" (CSWSS). The main clinical features were partial motor seizures, mental retardation and motor deficit. The EEG findings were characterized by nearly continuous (> 85%) diffuse slow spike and wave activity in two patients, and localized to one hemisphere in two other cases during non-REM sleep. The treatment was effective in improving the clinical seizures, but not the EEG pattern. We believe that this epileptic syndrome has been overlooked and routine sleep EEG studies on epileptic children may disclose more cases of CSWSS. PMID- 7487538 TI - Atypical clinical and electroencephalographic pattern in a patient with subacute sclerosing panencephalitis. AB - We describe an atypical clinical and electroencephalographic (EEG) pattern observed during the course of subacute sclerosing panencephalitis in a 14 year old boy. In this patient with a two weeks history of partial complex seizures, the atypical EEG pattern was characterized by an initial left temporal focus which evolved to periodic lateralized epileptiform discharges (PLEDs) and, only during the 3rd and 4th weeks the typical bilateral and generalized periodic complexes appeared. PMID- 7487537 TI - Complex partial status epilepticus in a child. AB - Complex partial status epilepticus (SE) has been reported rarely in children. We describe the clinical case of a 14 year-old girl with complex partial seizures (CPS) since age 10 who developed a complex partial SE probably because she was not adherent to treatment. The neurologic examination and computed tomography scan were normal. During the SE she received diazepam and phenytoin and became free of the seizures after 5 minutes. The ictal EEG showed spikes and slow waves over the right temporal region. PMID- 7487540 TI - [Reversible cerebral changes in hemolytic-uremic syndrome]. AB - The hemolytic-uremic syndrome is a pathology characterized by a triad consisting of acute renal failure, microangiopathic hemolytic anemia and thrombocytopenia, with complications of the central nervous system arising in a considerable number of cases. Altered cranial computerized tomography examinations usually reveal cerebral infarctions. We present here two cases in which diffuse hypodensity was observed in the white matter in addition to the infarcts. This hypodensity was reversible after resolution of the acute phase of the disease, as is also the case for the alterations described in uremic encephalopathy and in hypertensive encephalopathy of other etiologies. PMID- 7487539 TI - [Induced visual crisis and non-ketotic hyperglycemia: a case report]. AB - In a 47-years-old male patient, partial visual seizures triggered by movement was an early symptom of nonketotic hyperglycemia. Seizures were resistant to conventional anticonvulsivant therapy. During seizures, EEG showed discharges over the right hemisphere. CT was normal. A search for reflex seizures and hyperglycemia should be routinely carried out in patients with repeated focal visual seizures, as shown in the case reported. PMID- 7487542 TI - [Precocious Parkinson's disease associated with "eye-of-the-tiger" type pallidal lesions]. AB - We report the case of a 56-years-old woman patient, born to unrelated parents, who since 26-years-old gradually developed bradykinesia, rigidity, tremor of both hands, and speech and gait difficulties. Her past history was unremarkable. There was no family history of neurologic disease. She was admitted to our Hospital at age 39 and at that time she presented a full parkinsonian syndrome. The following tests were normal or negative: routine blood studies, serum copper, ceruloplasmin and cerebrospinal fluid examination. There was not Kayser-Fleicher ring, and fundoscopic examination revealed no abnormalities. Levodopa was introduced and response was good for more than ten years, despite early-onset of dyskinesias (three months after the introduction of the drug). After 30 years under levodopa she still presents a moderate response but with severe fluctuations of the motor performance. Except for slowness of cognition she developed no other neuropsychological impairments, and a recent neurological examination disclosed no abnormalities besides a parkinsonian syndrome. One year ago, a magnetic resonance imaging (MRI) was performed and showed bilateral, symmetrical lesions with "eye-of-the-tiger" pattern. This case illustrates the pathological heterogeneity of early-onset parkinsonism and suggests the possibility to find the typical MRI lesions seen in Hallervorden-Spatz disease in other degenerative affections involving globus pallidus. PMID- 7487543 TI - DOPA-sensitive progressive dystonia of childhood with diurnal fluctuations of symptoms: a case report. AB - Progressive dystonia with diurnal fluctuations sensitive to levodopa, also known as Segawa's disease, is a rare form of autosomal dominant extrapyramidal disease in the pediatric age group. The dystonic and Parkinson-like symptoms are the main clinical features of the disease and, characteristically but not in all cases, show a diurnal variation. They are absent or present to a lesser extent in the morning, worsening during the day. Treatment with small doses of levodopa results in remission or marked improvement of the symptomatology. We present the case of a 11 years old female patient that developed a dystonic posture in her feet that led her to a tip-toe walking pattern, since the age of 2. Diurnal fluctuations of the symptomatology were noticed by her mother. At 7 years of age she developed a left deviation of the head and an abnormal flexor posture of the left arm. In the next years the symptoms progressed and the fluctuations became less evident. At the age of 10, they were present soon after she woke up in the morning. The neurological examination disclosed a dystonic posturing of the head and left arm, a generalized rigidity of the extremities and a palpebral tremor. Laboratory examinations, including copper and ceruloplasmin, and neuro-imaging studies were negative. She was started on levodopa 150 mg/day with prompt disappearance of the symptomatology. After one-year follow-up she is symptom-free with only 100 mg/day of levodopa. No adverse effect was observed so far. PMID- 7487544 TI - [Multiple sclerosis simulating cerebral tumor: report of a case with histopathological confirmation]. AB - A 41 year old woman with subacute onset of moderate and progressive right hemiparesis showed on the MRI a unilateral white matter lesion on the left temporoparietal region and a small lesion on the right frontal region. The CSF was normal. Because possible diagnosis of tumor or abscess, the patient underwent a cerebral biopsy. The histological result was of brain tissue with demyelination and reactive changes. The stains for viral etiology were negative. PMID- 7487541 TI - [Acute neurogenic pulmonary edema: a case report]. AB - The authors report a case of acute neurogenic pulmonary edema in a 28-year-old woman who presented rupture of an internal carotid artery aneurysm and subarachnoid hemorrhage. The respiratory disorders started at the same time the patient's symptomatology aggravated. Some etiological and pathophysiological aspects on neurogenic pulmonary edema are revised. PMID- 7487545 TI - [Cerebellar abscess by Nocardia: a case report]. AB - The authors describe a case of cerebellar abscess by Nocardia in a patient with the acquired immunodeficiency syndrome (AIDS) that was submitted to a posterior fossa craniectomy for diagnosis and treatment. Pathological and neuroimage findings are discussed as well as the surgical approach taking into account literature data on the subject. PMID- 7487546 TI - Familial glioblastoma. AB - The authors describe a family with three members affected by glioblastoma. The proband patient, a 7 year-old girl, developed a rare complication, a pulmonary metastasis. Chromosomal analysis of her peripheral blood lymphocytes showed a normal karyotype (46, XX), without structural abnormalities. Cytogenetic study of the tumor cells disclosed several abnormalities: 46, XX, 7q-/46, XX, -2, 4p-, 7p , +15/46, XX. Some aspects about genetics of glial neoplasms are discussed. PMID- 7487547 TI - [The cutaneous extensor plantar reflex (Babinski, 1896/1898)]. AB - The extensor plantar reflex was described by Babinski in 1896. Given the obvious relevance of the sign for internal medicine as well as the paucity of translations of the original sources into Portuguese, we thought it timely to recall the ingenious arguments Babinski used to demonstrate his views on the "toe phenomenon", as he would call it. A careful analysis of Babinski's writings suggests, further, that he was driven by keen intuition as well as by medico legal interests. PMID- 7487548 TI - [Endocrine treatment of meningiomas: a review]. AB - Meningiomas are benign tumors of central nervous system. They have high rates of relapse and sometimes are not amenable to total removal mainly when involve vital structures. Clinical, epidemiological, biochemical and "in vitro" experiment show evidence that meningioma growth is influenced by steroid hormonal medium. Several clinical trials have explored these meningioma characteristics with the use of substances interfering with steroid actions: RU486 (antiprogestinic and antiglucocorticoid), medroxiprogesterone acetate (antiprogestinic), gestrinone (antiestrogenic e antiprogestinic), tamoxifen (antiestrogenic) and buserelin (LHRH superagonist), beyond of octreotide (somatostatin analog) and bromocriptine (dopaminergic agonist). Other substances have potential for the meningiomas treatment: aminogluthetimide, suramin and trapidil. In this review, we analyzed the literature about these aspects. PMID- 7487550 TI - The neurological adaptation of ICD-10 (ICD-10 NA). PMID- 7487549 TI - Energetic and spatial constraints of arterial networks. AB - The principle of minimum work (PMW) is a parametric optimization model for the growth and adaptation of arterial trees. A balance between energy dissipation due to frictional resistance of laminar flow (shear stress) and the minimum volume of the blood and vessel wall tissue is achieved when the vessel radii are adjusted to the cube root of the volumetric flow. The PMW is known to apply over several magnitudes of vessel calibers, and in many different organs, including the brain, in humans and in animals. Animal studies suggest that blood flow in arteries is approximately proportional to the cube of the vessel radius, and that arteries alter their caliber in response to sustained changes of blood flow according to PMW. Remodelling of the retinal arteriolar network to long-term changes in blood flow was observed in humans. Remodelling of whole arterial networks occurs in the form of increase or diminishing of vessel calibers. Shear stress induced endothelial mediation seems to be the regulating mechanism for the maintenance of this optimum blood flow/vessel diameter relation. Arterial trees are also expected to be nearly space filing. The vascular system is constructed in such a way that, while blood vessels occupy only a small percentage of the body volume leaving the bulk to tissue, they also crisscross organs so tightly that every point in the tissue lies on the boundary between an artery and a vein. This review describes how the energetic optimum principle for least energy cost for blood flow is also compatible with the spatial constraints of arterial networks according to concepts derived from fractal geometry. PMID- 7487551 TI - [Pattern visual evoked potentials in migraine subjects without aura]. PMID- 7487553 TI - Psychiatric classification of nonconversion nonepileptic seizures. PMID- 7487554 TI - Genotype-phenotype correlations in human skeletal muscle sodium channel diseases. PMID- 7487552 TI - Snoring and headache: an important association. PMID- 7487555 TI - Managed care in JAMA and the Archives journals. A call for papers for coordinated theme issues. PMID- 7487556 TI - Successful treatment of neuropathies in patients with diabetes mellitus. AB - OBJECTIVES: To report and characterize two forms of disabling progressive peripheral neuropathy in patients with diabetes mellitus, which respond to anti inflammatory and/or anti-immune treatment. DESIGN: Review of clinical, electrophysiologic, and pathologic findings and results of treatment. SETTING: University medical center. PATIENTS: Twenty-one patients with diabetes mellitus to whom we gave anti-inflammatory and/or anti-immune treatment for progressive peripheral neuropathy during the past 6 years. MAIN OUTCOME MEASURES: Patients were interviewed and examined at intervals before and after beginning treatment with intravenous immunoglobulin (n = 15), prednisone (n = 13), cyclophosphamide (n = 5), plasma exchange (n = 3), and azathioprine (n = 1) (alone or in combination). RESULTS: Fifteen patients had evidence of axonal neuropathy by electrophysiologic studies (group A). All 15 patients had non-insulin-dependent diabetes mellitus, 10 patients had weight loss, and 13 patients had prominent involvement of thighs and/or thoracic bands consistent with diabetic amyotrophy or mononeuropathy multiplex. Small vessel disease was seen in all 10 patients who underwent biopsy, with perivascular or vascular inflammation seen in seven patients. Six patients had demyelinating neuropathy by electrophysiologic criteria (group B). All these patients had insulin-dependent diabetes mellitus, and no one had weight loss. The process was asymmetric in three patients and involved thoracic or abdominal regions in two patients. Onion bulbs were seen in all four patients who underwent biopsy, but no vascular inflammation or occlusion was seen. In all patients in both groups, worsening of their conditions stopped and improvement started after beginning treatment. CONCLUSION: Neuropathies responsive to anti-inflammatory and/or anti-immune therapy in patients with diabetes mellitus include (1) multifocal axonal neuropathy caused by inflammatory vasculopathy, predominantly in patients with non-insulin-dependent diabetes mellitus, indistinguishable from diabetic proximal neuropathy or mononeuropathy multiplex, and (2) demyelinating neuropathy indistinguishable from chronic inflammatory demyelinating polyneuropathy, predominantly in patients with insulin dependent diabetes mellitus. PMID- 7487557 TI - The value of PET scan (and MRI and Wada test) in patients with bitemporal epileptiform abnormalities. AB - BACKGROUND: Lateralization remains difficult in patients with bitemporal epileptiform abnormalities. OBJECTIVE: To evaluate the reliability of the interictal fluorodeoxyglucose F18-positron emission tomographic (PET) scan in this setting. METHODS: We analyzed PET scan findings in 25 patients who required bilateral depth electrodes for lateralization of temporal lobe epilepsy. The contribution of magnetic resonance imaging and the intracarotid amobarbital procedure was also analyzed. RESULTS: Positron emission tomographic scan revealed lateralized hypometabolism in 15 patients. This was in agreement with the depth evaluation in 10 cases (67%). In five cases (33%), the PET and depth findings were in disagreement, with no predominant side of seizure origin by depth electroencephalography (EEG) in four cases and PET lateralization opposite to depth findings in one case. In 10 of the 25 patients, PET scans could not lateralize hypometabolism to one temporal lobe, of which six were confirmed by depth EEG not to have a predominant side of seizure onset. The overall sensitivity of PET for concordant lateralized seizure onset was 67%. Ten of 14 patients who underwent a lobectomy had a successful outcome, including three of four patients whose temporal lobe epilepsy was lateralized by depth EEG but not by PET. One patient had PET lateralization opposite to the depth EEG and side of surgery and failed to improve. CONCLUSIONS: In this highly selected group of patients, the lateralizing value of PET and magnetic resonance imaging was somewhat diminished, and the intracarotid amobarbital procedure was not found useful for lateralization. Positron emission tomography and magnetic resonance imaging, when pointing to the side opposite depth EEG, may indicate a poor prognosis. PMID- 7487558 TI - Scapuloperoneal syndromes. Absence of linkage to the 4q35 FSHD locus. AB - OBJECTIVE: To investigate whether two forms of the scapuloperoneal syndrome result from genetic defects allelic to facioscapulohumeral dystrophy (FSHD). DESIGN: Two kindreds with scapuloperoneal syndromes underwent clinical, histologic, and electrophysiologic evaluation followed by genetic evaluation with probes closely linked to FSHD. RESULTS: Although the proband in each kindred had facial, scapular stabilizer, and humeral weakness, raising the possibility of FSHD, evaluation of multiple other affected family members showed patterns of involvement that were clinically distinct from typical FSHD. In addition, DNA studies showed no linkage to the 4q35 FSHD locus in either kindred. CONCLUSION: We conclude that these two forms of the scapuloperoneal syndrome are genetically distinct from FSHD. PMID- 7487559 TI - Apolipoprotein E epsilon 4 allele and the lifetime risk of Alzheimer's disease. What physicians know, and what they should know. AB - BACKGROUND: Published studies now show a clear association between Alzheimer's disease (AD) and the apolipoprotein E epsilon 4 allele (APOE* epsilon 4). The clinical value of this information to estimate a healthy individual's lifetime risk of AD has not been well delineated. Physicians dealing with AD may not know either the lifetime risk of developing AD or the effect of the APOE genotype on this risk. Because the lifetime risk of AD depends in part on life expectancy, and available figures on APOE are not population based, a computation is necessary to derive risk estimates useful to physicians. OBJECTIVES: To estimate the lifetime risk of AD and the effect of APOE genotype information on that risk and to assess the knowledge of these risks among physicians who manage patients with dementia. DESIGN: Estimation of risk of AD and survey of physician awareness. The lifetime risk of developing AD without APOE genotype information was first computed for 65-year-olds from existing epidemiologic studies of age related AD incidence and an actuarial life-table analysis. Using this computed a priori risk of AD and published studies of APOE genotypes in individuals with and without AD, we used a Bayesian analysis to determine the risk of developing AD, with and without an APOE* epsilon 4 allele, for unaffected 65-year-olds. To assess physician knowledge of the lifetime risk of AD and the effect of APOE genotyping on the risk, 50 neurologists, internists, geriatricians, geriatric psychiatrists, and family physicians who manage patients with dementia were randomly selected to participate in a questionnaire-driven telephone survey. RESULTS: In a person with no family history of AD, the epidemiologic/actuarial lifetime risk of AD is approximately 15%. Based on a Bayesian calculation and published APOE data, the lifetime risk of AD is 29% for individuals with one APOE* epsilon 4 allele and it is 9% if no APOE* epsilon 4 allele is present. Physician awareness survey results were as follows: 42% of physicians correctly estimated the approximate lifetime risk of AD; of these, only one third were moderately sure of their response. Only three physicians correctly estimated the change in risk given the APOE* epsilon 4 genotype; only one of these was at least moderately sure. CONCLUSIONS: Determining the APOE* epsilon 4 status of healthy adults with no family history of AD approximately doubles (for the epsilon 4 allele) or reduces by 40% (for the non-epsilon 4 allele) the uninformed lifetime risk of developing AD. Even with an APOE* epsilon 4 allele, the lifetime risk remains below 30%. Most physicians managing patients with AD do not know the lifetime risk of AD, and very few know how APOE* epsilon 4 status modifies the risk. These clinically relevant risk figures should be more widely disseminated among physicians. PMID- 7487560 TI - Plasma and red blood cell thiamine deficiency in patients with dementia of the Alzheimer's type. AB - OBJECTIVES: To determine the prevalence of plasma thiamine deficiency in patients referred to a memory disorder clinic and to compare plasma thiamine levels with red blood cell (RBC) thiamine levels. To determine if patients with senile dementia of the Alzheimer's type (SDAT) differ from those without SDAT in either plasma or RBC thiamine levels. DESIGN: Case-control study. SETTING: Ambulatory care referral center. PATIENTS: Consecutive sample of 34 patients; 17 patients who met the National Institute of Neurological and Communicative Disorders and Stroke-Alzheimer's Disease and Related Disorders Association criteria for probable Alzheimer's disease and 17 patients with other forms of dementia. METHODS: Plasma and RBC thiamine levels were determined in all patients with the use of a microbiologic assay known for its specificity to biological forms of thiamine. Vitamin supplementation was determined by chart review. OUTCOME MEASURES: Plasma and RBC thiamine levels. RESULTS: Patients with SDAT were found to have significantly lower plasma thiamine levels than patients without SDAT. Low plasma thiamine levels were detected in a significantly larger proportion of patients with SDAT than in patients without SDAT. Red blood cell thiamine levels did not correlate with the clinical diagnosis of SDAT. Vitamin supplementation did not correlate with diagnosis and plasma or RBC thiamine levels. CONCLUSIONS: A significant proportion of patients with SDAT may have a thiamine deficiency, which may have an impact on cognitive function. Currently used assays may not be adequate to assess thiamine status. PMID- 7487561 TI - Cognitive test performance in detecting, staging, and tracking Alzheimer's disease. AB - OBJECTIVES: To identify the specific cognitive deficits that characterize Alzheimer's disease (AD) and determine which cognitive tests, or combination of tests, are best for detecting AD (ie, distinguishing patients with AD from normal control subjects), staging AD (ie, distinguishing different severities of dementia), and tracking disease progression. SUBJECTS: Patients with AD (n = 123) and normal control subjects (n = 60) of comparable age, education, and gender distribution. SETTING: Outpatient care. MEASURES: Ten cognitive tests of memory, language, visuospatial abilities, and reasoning; the Information, Memory and Concentration subtest of the Blessed Dementia Scale, and the total score on an activities of daily living questionnaire. DESIGN: Patients with AD were tested every 6 to 24 months over a span of up to 5.5 years. RESULTS: Patients with AD were significantly inferior to normal control subjects on all cognitive tests. The scores of patients with AD worsened over time. Delayed recall of stories and figures showed sharp deterioration to an early floor, consistent with the finding that these tests discriminated patients with mild AD from normal control subjects well but were poor for staging. Confrontation naming, semantic fluency, and immediate recognition of geometric figures showed steady linear decline across time for patients with AD, consistent with these tests being found best for staging dementia severity. CONCLUSIONS: We postulate that the pathologic bases of impairment in delayed recall are atrophy of cholinergic ventral forebrain neurons and partial deafferentation of the hippocampus, both of which occur early in the course of AD. Worsening language and visuospatial abilities likely reflect progressive loss of neocortical neurons and their connections. PMID- 7487562 TI - Thrombosis of the deep venous drainage of the brain in adults. Analysis of seven cases with review of the literature. AB - OBJECTIVES: To characterize the clinical, radiographic, and pathologic findings in thrombosis of the deep venous drainage of the brain. To highlight clinical and radiographic findings that may lead to the diagnosis of disease and distinguish it from dural sinus thrombosis. To review the published literature on this disorder. DESIGN: Retrospective review of the medical and radiographic records of seven patients from three institutions over the past 10 years. Review of the English language literature from 1971 to the present. RESULTS: All seven patients had risk factors for cerebral venous thrombosis. Five patients presented with a short, rapidly progressing course characterized by headache, nausea and vomiting, and decline in level of consciousness. All five patients died or were rendered severely disabled. Computed tomographic scans, magnetic resonance imaging, and magnetic resonance angiography showed findings associated with deep cerebral vein thrombosis in three of four, in five of five, and in three of three patients, respectively. Transfemoral catheter angiography was diagnostic in two of two patients. Twenty-one reported cases of deep cerebral venous thrombosis were identified in the literature. CONCLUSIONS: When the two populations are combined and compared with large series of patients with dural sinus thrombosis, patients with deep venous system thrombosis are more commonly women, tend to present with a more rapidly declining time course, altered consciousness, and long tract signs. Death or long-term sequelae are far more likely to occur in internal cerebral vein thrombosis than with dural sinus thrombosis. Unenhanced computed tomography can demonstrate findings that are strongly suggestive of the diagnosis. Magnetic resonance imaging and magnetic resonance angiography are confirmatory. Angiography may still be necessary when the diagnosis is not clear. PMID- 7487564 TI - Conditions that mimic stroke in the emergency department. Implications for acute stroke trials. AB - OBJECTIVE: To determine if any clinical variables allow early discrimination between stroke and other conditions presenting with a strokelike picture. BACKGROUND: New therapeutic modalities for the treatment of acute ischemic stroke are under active investigation. Many of these treatments have potential adverse effects. It is well known that noncerebrovascular conditions can present with a clinical picture mimicking stroke, so early accurate differentiation of such "mimics" from true stroke is essential. METHODS: Consecutive patients who presented to the emergency department with an initial diagnosis of stroke between January 1990 and January 1992 were evaluated. Chart review allowed these patients to be classified into two final diagnostic groups: stroke mimic and true stroke. Logistic regression was used to estimate the effects of predictor variables measured at initial evaluation on the final diagnosis. RESULTS: There were 411 patients initially diagnosed as having stroke. Of these, 78 patients (19%) were eventually found to have mimics, the majority comprising postictal states, systemic infections, tumors, and toxic-metabolic disturbances. Univariate analysis showed that decreased level of consciousness and normal eye movements increased the odds of mimic, while abnormal visual fields, diastolic blood pressure greater than 90 mm Hg, atrial fibrillation on electrocardiogram, and history of angina decreased the odds of mimic. Multivariate analysis showed that decreased consciousness increased, while history of angina decreased, the odds of mimic. Sensitivity of this model for predicting mimics was only 21% while specificity was 96%. CONCLUSION: For the neurologist faced with an immediate decision as to whether to randomize a patient with probable stroke to an acute treatment protocol, these findings have some usefulness but mandate further research into conditions that mimic stroke in the emergency department. PMID- 7487565 TI - Profile of a neurology residency. AB - The pattern and frequency of patient encounters during the Boston (Mass) University adult neurology residency program (1988 to 1991) for one resident was compared with that in general neurology practice as well as with the frequency of neurologic disorders in the US population. A total of 1332 new patients (85% adult, 15% pediatric) were seen during a 3-year period. This total represented 970 inpatients (73% of all patients) and 362 outpatients (27%). The resident encountered more patients in the hospital (7.5 admissions or 13 consultations per week) and fewer patients in the clinic (2.5 new outpatients per week) than does the average community neurologist (two admissions, 8.7 consultations, and 13.2 new outpatients). The most common diagnosis for an admission encounter was acute ischemic infarct; for a consultation, metabolic encephalopathy; and for an outpatient encounter, radiculopathy. Less prevalent neurologic disorders in the United States (eg, cognitive, demyelinating, movement, and neoplastic disorders) were encountered more frequently in residency than were very prevalent neurologic disorders (eg, headache and trauma). This is the first reported summary of all patients one resident actually encountered during neurology training. The patient encounter profile suggests that this residency training overemphasized acute inpatient care of less prevalent neurologic disorders compared with outpatient care of more prevalent disorders commonly seen in a neurology practice. Accumulation of similar data from other residencies and practicing neurologists can help residency directors assess the changing needs of residents in training and guide curriculum in response to changes in practice patterns. PMID- 7487566 TI - Streptococcus oralis previously identified as uncommon 'Streptococcus sanguis' in Behcet's disease. AB - The relation between the biochemical and serological properties of 'Streptococcus sanguis' was studied to characterize the strains isolated from dental plaque of patients with Behcet's disease and controls. Seven reference and 100 clinical strains preserved by the Behcet's Disease Research Committee of Japan were identified using established criteria and differentiated with antisera against Strep. oralis ATCC 10557, Strep. sanguis ATCC 10556 and 'Strep. sanguis' ST7, compatible with the criteria. Uncommon serovars (serotypes) KTH-1 (= ATCC 49298), KTH-2 (= ATCC 49296), KTH-3 (= ATCC KTH-4 (= ATCC 49297) and B220 (serovar KTH-1) with both IgA1 protease and neuraminidase (sialidase) were identified as Strep. oralis, whereas common serovars ST3 with IgA1 protease alone and ST7 without both enzymes were identified as Strep. sanguis and Strep. gordonii, respectively. Isolates previously ranked as uncommon serovars were identified as Strep. oralis, whereas the rest ranked as common serovars were identified as the same species as those of the grouping strains. A soft-agar technique was available for species identification except for Strep. oralis serovar KTH-1 reacting with the antiserum against Strep. gordonii ST7. The frequency of isolation of Strep. oralis was higher in Behcet's disease (52%) than in controls (38%), but no difference was observed between the properties of the two groups of isolates. Strep. oralis virulence factors may be involved in breach the mucosal barrier in patients with specific reactivity to these antigens and inducing Behcet's disease. PMID- 7487563 TI - Delayed latency of the event-related brain potential P3A component in HIV disease. Progressive effects with increasing cognitive impairment. AB - OBJECTIVE: To examine the degree to which P3A latency was sensitive to the early and progressive effects of human immunodeficiency virus (HIV) disease on frontal cortex function by studying HIV-positive subjects who varied in degree of cognitive impairment. DESIGN: Event-related brain potential studies of four groups of subjects: cognitively nonimpaired high-risk HIV-negative subjects, cognitively nonimpaired HIV-positive subjects, cognitively mildly to moderately impaired HIV-positive subjects, and cognitively severely impaired HIV-positive subjects. SETTING: Voluntarily participating subjects on an outpatient basis at a medical center facility. PARTICIPANTS: Seventy-one community-residing gay or bisexual HIV-positive male volunteers were compared with 17 HIV-negative male gay or bisexual subjects used as a control sample. The HIV-positive subjects were stratified with regard to severity of cognitive impairment into the following three subsamples: subjects who were cognitively normal (n = 35), subjects with mild to moderate cognitive impairment (n = 20), and subjects with severe cognitive impairment (n = 16), with the samples closely matched in age. The HIV positive subsamples were closely matched on percentage of CD4 lymphocytes. Subjects were excluded if they reported a history of drug or alcohol abuse, a major mental disorder, a head injury with loss of consciousness, or brain disease other than HIV related. MAIN OUTCOME MEASURE: P3A latency. RESULTS: P3A latency was significantly delayed in HIV-positive subjects compared with HIV-negative control subjects, with a delay of 12 milliseconds in the cognitively normal group (P < .02) and the magnitude of delay increasing with increasing severity of HIV associated cognitive impairments (P < .001). Delayed P3A was primarily associated with the progression of HIV-associated cognitive impairment, with a secondary and additive association with severity of HIV-associated medical illness. CONCLUSION: This finding suggests that delayed P3A latency is sensitive to the relatively early central nervous system effects of HIV and progresses with worsening of the central nervous system effects of HIV. PMID- 7487567 TI - Rate of oxygen consumption by parotid atrophic acinar cells from rats fed liquid diet. AB - Adult male rats were maintained on a nutritionally adequate liquid-diet or laboratory chow and water (control), for 9 days. They were then killed and the parotid glands removed. Enzymatically dispersed acinar-cell preparations were used to study rates of oxygen consumption (QO2) using a Clark oxygen electrode. For both CON and LD cell preparations the basal QO2 was 0.132 (+/- 0.03-0.05) nmol O2 per microgram DNA per min and in each case this was increased approx. 2.4 fold under stimulation by carbachol (10 microM) and approx. 3.1-fold by adrenaline (10 microM). Isoprenaline (10 microM) elicited no significant increase in QO2. Addition of ouabain (2.5 mM) or removal of Ca2+ from the extracellular medium prevented the action of carbachol to increase QO2 over a sustained period. There were no significant differences in the basal, agonist-stimulated, or ouabain-sensitive QO2 by dispersed acinar cells from liquid-diet rats compared to control. The results are consistent with the notion that liquid diet-induced atrophy represents a physiological adaptation rather than a pathological change. Nevertheless, they also indicate that in the acinar cells from liquid-diet rats the Na+/K+ ATPase is required to operate at normal levels of energy consumption despite the reduced acinar-cell volume and consequently lower levels of agonist elicited transepithelial ion movements known to occur in these cells. PMID- 7487568 TI - The influence of different fluoride salts on fluoride-mediated inhibition of peroxidase activity in human saliva. AB - Fluoride-mediated inhibition of peroxidase potential activity in human saliva was investigated using NaF, NH4F, CaF2, Na2PO3F (MFP), SnF2 and TiF4. At pH 5.5 and for a 20 mM F concentration, the inhibition percentages increased from 2% for MFP and 5% for CaF2 to 61% for NaF and 65% for NH4F, while a 100% inhibition was observed at 10 mM for TiF4 and at 5 mM for SnF2. The inhibition was enhanced at acid pH and removed at pH 7. The inhibition could be attributed to ionized F-, except for SnF2 and TiF4, in which part of the inhibition could be imputed to the cations. PMID- 7487569 TI - The flow rate and electrolyte composition of whole saliva elicited by the use of sucrose-containing and sugar-free chewing-gums. AB - On two occasions, 12 adults collected unstimulated saliva and then eight samples of saliva over a 20-min period while chewing 3 g of either Wrigley's Spearmint sucrose-containing gum (SCG) or sugar-free gum (SFG) at 70 chews/min. The flow rates peaked initially, then fell with duration of stimulation. With the SFG they were slightly but significantly higher than with the SCG after 4 min of chewing. The sum of the concentrations of cations minus the sum of the concentrations of anions was not significantly different from zero for saliva elicited by the SCG. However, for unstimulated saliva and that elicited by SFG, there was a slight positive anion balance. A second series of saliva collections with SCG and SFG was made by the same 12 participants and these samples were analysed for lactate. For these collections the flow rates with SCG were not significantly less than with the SFG. The lactate concentration in saliva elicited by SCG peaked at 1.82 mmol/l in samples collected over 8-15 min, whereas samples of saliva elicited by SFG had a mean lactate concentration of 0.21 mmol/l. Of the lactate formed during the metabolism of sucrose by the oral bacteria, only 2% or less appeared to be derived from the metabolism of micro-organisms free in saliva, the balance presumably being formed in dental plaque and entering the saliva by diffusion. All saliva samples were supersaturated with respect to hydroxyapatite but stimulated saliva was significantly more supersaturated than unstimulated saliva.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7487570 TI - Morphological analysis of dentine formation in young rat molars during the recovery phase with calcium alone or combined with xylitol following a low calcium dietary regimen. AB - The effects of dietary calcium deficiency and subsequent replenishment of the diet with calcium alone or with xylitol were studied. Thirty 3-week-old Wistar rats were labelled with an i.p. tetracycline injection. Twenty rats were fed a diet with 0.026% calcium (Ca-deficient); 10 received a 0.5% Ca diet (controls). After 3 weeks the tetracycline labelling was repeated. Replenishment of the diet was introduced for Ca-deficient rats, and 10 received additional 5% xylitol in the diet. After 4 weeks the labelling was repeated and the animals were decapitated. Dentine formation was measured by the tetracycline stripes in the lower first and second molars. Calcium deficiency during the first 3 weeks reduced dentine formation. In the control and xylitol groups, a much smaller amount of dentine was formed during the recovery period. With Ca alone, dentine formation was faster than in the controls or Ca-xylitol group and did not differ from the Ca-deficient period. These results indicate that in rat molars the odontoblasts can accelerate the rate of dentine formation when calcium is restored to the diet, at least during primary dentinogenesis. This effect was not seen when 5% xylitol was added to the replenishment diet. PMID- 7487571 TI - The dephosphorylation of 22-kDa phosphoprotein by type 2B protein phosphatase in rat parotid acinar cells. AB - In saponin-permeabilized rat parotid acinar cells, cyclic AMP and 3-isobutyl-1 methylxanthine stimulated the phosphorylation of three particulate proteins with molecular masses of 34, 26 and 22 kDa. The particulate fractions containing 22 kDa phosphoprotein were isolated from the cells labelled with [gamma-32P]ATP and used to study the dephosphorylation of the 22-kDa phosphoprotein. When the labelled fractions were incubated at 30 degrees C in the presence of 0.3 mM CaCl2 and 10 micrograms calmodulin, dephosphorylation of the 22-kDa phosphoprotein was evoked. Further addition of the type 2B phosphatase (Ca2+/calmodulin-stimulated protein phosphatase purified from bovine brain) resulted in a remarkable dephosphorylation of the 22-kDa phosphoprotein. Western immunoblotting showed that type 2B protein phosphatase exists in rat parotid acinar cells. These results suggest that type 2B protein phosphatase in those cells is involved in the dephosphorylation of the 22-kDa phosphoprotein. PMID- 7487572 TI - Time-related changes induced by zinc-deficient diet in the concentration of rat cheek epithelial membrane-coating granules. AB - Weanling male Sprague-Dawley rats were fed a diet containing 0.4 parts/10(6) zinc and controls were fed an identical diet supplemented with 40 parts/10(6) zinc. After 9, 18 and 27 days of zinc deficiency, specimens were excised from cheek epithelium and processed for transmission electron microscopy to study the concentration of membrane-coating granules (MCG). Their concentration was increased in the granular-cell layers of the zinc-deficient epithelium and became significantly greater after 18 and 27 days than 9 days of deficiency. MCGs appeared in the parakeratinized layers of zinc-deficient epithelium and their concentration became significantly greater after 27 days in comparison with 9 and 18 days of deficiency. Thus the intracellular retention of MCGs was increased in the granular and parakeratinized layers with the increase in time of zinc deficiency. PMID- 7487573 TI - Measurements of human minor salivary gland secretions from different oral sites. AB - Unstimulated secretions from labial (upper and lower), buccal, lingual and palatal minor glands were measured with a Periotron 6000 model 2 (Proflow) in 99 non-medicated, healthy individuals aged from 17 to 81 yr (49 males and 50 females). The palatal minor glands had the lowest flow rates, buccal and anterior lingual glands the highest, and labial glands (upper and lower) intermediate secretory rates. Lower labial glands (p < 0.01) in both sexes and upper labial glands in females (p < 0.05) demonstrated a significant decrease in unstimulated output with age, while buccal glands in both sexes (p < 0.05) showed a significant decrease in flow rates only after the age of 50 yr. Thus minor glands show regional variation in their secretory rates and labial and buccal glands appear to show a decreased unstimulated flow in older individuals. PMID- 7487574 TI - Comparison of host tissue and bacterial dipeptidyl peptidases in human gingival crevicular fluid by analytical isoelectric focusing. AB - Earlier work has shown that gingival crevicular fluid (GCF) contains dipeptidyl peptidase (DPP) activities that resemble those in host tissue. Here, further comparisons were made with enzymes from suspected periodontal pathogens. Gingival tissue and GCF were collected from patients with chronic periodontitis. DPP II and DPP IV fractions with acid and alkaline pH optima, respectively, were separated from crude tissue extracts by gel-filtration chromatography. Bacterial cell sonicates were prepared from broth cultures of reference strains. There was moderate to strong DPP activity with Capnocytophaga spp., Porphyromonas gingivalis and Prevotella spp., very weak activity with Treponema denticola and no detectable activity with Actinobacillus actinomycetemcomitans or Fusobacterium nucleatum. Banding patterns in GCF, tissue and bacterial samples were compared on substrate-impregnated overlay membranes applied to isoelectric focusing gels. In gels washed with acid buffer, GCF had bands corresponding to tissue DPP II. Use of an alkaline washing buffer showed GCF activity which closely matched tissue DPP IV that had been pretreated with neuraminidase, an enzyme found by others in the gingival crevice. P. Gingivalis gave multiple bands and several of these had counterparts in GCF. The apparent presence in GCF of the DPP from P. gingivalis is consistent with the association of this organism with destructive periodontitis. PMID- 7487576 TI - A comparative study of glucose and galactose uptake in pure cultures of human oral bacteria, salivary sediment and dental plaque. AB - The ability to utilize glucose and the weaker sugar acidogen, galactose, was surveyed in salivary sediment, pooled dental plaque, and in pure cultures of the bacteria that numerically comprise most of the bacteria in these mixed microbial systems. Except for a veillonella isolate, which showed no uptake of either sugar, glucose was utilized more rapidly than galactose by the 27 pure cultures tested and by both sediment and plaque. This sugar difference was also seen for two other measures of glycolysis, formation of acid and previously studied ability to produce an acidic pH. Rates of uptake of the two sugars by individual pure cultures varied considerably. Generally, the Gram-positive bacteria utilized glucose and galactose at rates similar to those seen with salivary sediment and dental plaque, whereas the Gram-negative cultures tested showed much slower uptakes. Bacteria previously identified as arginolytic had lower glucose and galactose uptake rates than similar non-arginolytic micro-organisms. This, together with the ability to produce base from arginine, would explain their tendency to produce a less acidic pH. In pure culture mixtures, uptakes were generally predictable and indicated an averaging effect. When the microbial compositions of salivary sediment or dental plaque were altered by mixing with pure cultures of high glucolytic activity, such as many of the Gram-positives, glucose uptake was enhanced. The opposite was observed when the less glucolytic Gram-negative bacteria were similarly incorporated. As well as determining the glucose and galactose uptake rates of the various bacteria that collectively comprise the bulk of the salivary sediment and supragingival plaque microfloras, this study has shown how variation in microbial composition affects sugar uptake rates and has indicated how microbial composition could be manipulated to produce dental plaques with different capacities to ferment sugars and presumably different cariogenicities. PMID- 7487575 TI - Effect of marginal ascorbic acid deficiency on saliva level of cortisol in the guinea pig. AB - Male guinea pigs subjected to prolonged marginal ascorbic acid deficiency developed moon facies and oedema, features of functional adrenal hypercorticism. Compared with age- and sex-matched controls fed an adequate diet for a similar period, ascorbate deficiency had no effect on submandibular gland weight but elicited a significant (p < 0.005) reduction in stimulated whole-saliva flow rate. Plasma cortisol concentration (nmol/L) was significantly increased (p < 0.005) in the deficient animals (998.21 +/- 57.19 compared to 254.66 +/- 15.62 for the controls). Associated with marked hypercortisolaemia in the deficient animals was a significant (p < 0.01) but less prominent increase in the whole saliva cortisol level, resulting in a mean saliva/plasma cortisol ratio of 46% for this group compared to 72% for the controls. Increased corticosteroid levels suppress immunological and inflammatory responses, particularly neutrophil function, impair production of some cytokines, inhibit collagen synthesis, and impair wound healing and bone matrix formation. Numerous conditions such as ageing, stress, smoking, ionizing radiation, ingestion of drugs, protein malnutrition, diabetes, and several other pathological states, which are among the risk factors for xerostomia and periodontal/oral mucosal lesions, promote tissue depletion of ascorbate. This study suggests that increased salivary and blood levels of glucocorticoids in these conditions may be important in reducing the ability of the host to mount an effective immune response to oral pathogens. PMID- 7487577 TI - Bacteria in human mouths involved in the production and utilization of hydrogen peroxide. AB - Earlier studies have demonstrated that pure cultures of oral streptococci produce hydrogen peroxide but none has found any free peroxide in dental plaque or salivary sediment despite streptococci being major components of their mixed bacterial populations. The absence of peroxide in plaque and sediment could be due to the dominance of its destruction over its formation by bacterial constituents. To identify which of the oral bacteria might be involved in such a possibility, pure cultures of 27 different oral bacteria were surveyed (as well as dental plaque and sediment) for their peroxide-forming and peroxide-removing capabilities. Peroxide production was measured for each of the pure cultures by incubation with glucose at low and high substrate concentrations (2.8 and 28.0 mM) for 4 h and with the pH kept at 7.0 by a pH-stat. Removal of hydrogen peroxide was assessed in similar experiments where peroxide at 0, 29.4, 147.2 or 294.4 mM [0, 0.1, 0.5 and 1% (w/v)] replaced the glucose. Hydrogen peroxide formation was seen with only three of the bacteria tested, Streptococcus sanguis I and II (sanguis and oralis), and Strep. mitior (mitis biotype I); levels of hydrogen peroxide between 2.2 and 9.8 mM were produced when these micro-organisms were grown aerobically and 1.1 and 3.9 mM when grown anaerobically. Earlier reports indicate that such levels were usually sufficient to inhibit the growth of many plaque bacteria. The amounts formed were similar at the two glucose levels tested, suggesting that maximum peroxide production is reached at low glucose concentration. None of the three peroxide-producing organisms was able to utilize hydrogen peroxide but five of the other 24 tested, Neisseria sicca, Haemophilus segnis, H. parainfluenzae, Actinomyces viscosus and Staphylococcus epidermidis, could readily do so, as could the mixed bacteria in salivary sediment and dental plaque, both of which contain relatively high numbers of these peroxide-utilizing micro-organisms. The ability of the bacteria in plaque and sediment to degrade hydrogen peroxide was considerable and extremely rapid; peroxide removal and usually complete within the first 15 min of the incubation even when its initial level was as high as 294.4 mM. This almost overwhelming ability to remove peroxide was confirmed when peroxide-producing and -using cultures were mixed and when each of eight salivary sediments was incubated with glucose and with peroxide at concentrations up to 294.4 mM. In the glucose incubations, no hydrogen peroxide was observed, indicating dominance of microbial peroxide removers over hydrogen peroxide producers.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 7487579 TI - The effects of blindfolding and blindness on the unstimulated and chewing-gum stimulated flow rates of whole saliva. AB - In the early 1970s, Shannon and his colleagues reported that blindfolding caused a reduction in the flow rate of unstimulated and stimulated parotid and submandibular saliva. A study in three parts was now made to investigate the effects of blindfolding or blindness on the flow rate of whole saliva. For the first study, unstimulated whole saliva (UWS) was collected for 5 min from 34 individuals and then eight samples of chewing-gum-stimulated whole saliva (SWS) over a 20-min period. This was carried out on four separate occasions, on two of which, in random order, the individuals collected the saliva immediately after being blindfolded. For the second study, 33 of the individuals repeated the saliva collection protocol on two occasions, with and without blindfolding, but with an accommodation period of 20 min after blindfolding before beginning collection of UWS and with only three samples of SWS being collected over a 4-min period. The flow rate of UWS while blindfolded was significantly (p < 0.0001) reduced (to 64 and 71% of the flow rate while sighted, in the first and second studies, respectively), as was the flow rate of SWS (to 81-86%, p < 0.0001; and 91-95%, p = 0.0014, respectively). For the third study, UWS and SWS were collected from 24 blind individuals, ranging in age from 13-73 years, and from 24 age- and gender-matched controls. The flow rates of UWS and SWS were not significantly different in the blind participants and controls.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7487580 TI - Disaccharide analysis of chondroitin sulphate in human gingival crevicular fluid using high-performance liquid chromatography. AB - Gingival crevicular fluid (GCF) was collected into capillary tubes from healthy gingiva and sites of advanced periodontitis. Following digestion with Pronase E, the glycosaminoglycans were isolated by successive precipitation into 5% cetylpyridinium chloride and 95% ethanol. Unsaturated disaccharide isomers of chondroitin sulphate, obtained following chondroitinase ACII digestion, were analysed by high-performance liquid chromatography. Chondroitin sulphate was found in all GCF samples, with greater amounts in patients with periodontal disease than at control sites with a relatively healthy periodontium. The predominant isomer in the periodontal diseased group was delta Di-4S, while that in the control group and serum samples was delta Di-0S. Comparison of the relative proportions of the unsaturated disaccharides in GCF with previously reported values for alveolar bone, cementum, gingiva and periodontal ligament, as well as for serum, indicates that the chondroitin sulphate present in GCF of patients with periodontal disease originated from the mineralized connective tissues of the periodontium, notably alveolar bone, possibly with some contributions from soft connective tissues of gingiva and periodontal ligament and from serum. PMID- 7487578 TI - The occurrence of silicon-containing particles in calcified lesions of DBA/2NCrj mice. AB - The crystalline structure in calcified deposits of DBA/2 mice was analysed, because understanding this is essential in assessing the significance of this species as an animal model for spontaneous soft-tissue mineralization. In order to remove the thin soft tissue which covers the lesion, the samples were bombarded with an argon beam. The uncoated samples were analysed with an electron probe microanalyser. The deposits were composed of a calcium phosphate material. At a higher magnification, smaller particles surrounding these large deposits were found. There were three types of particles: rod-shaped (calcium phosphate), polygonal (silicate) and round (about 1 microns dia). PMID- 7487581 TI - Estimation of the total saliva volume produced per day in five-year-old children. AB - Fifteen boys and 15 girls were asked to record for 2 days the time spent awake, eating meals or snacks, and sleeping. The salivary flow rates elicited by chewing foods were also determined. The mean flow rate (+/- SD) of unstimulated saliva was 0.26 +/- 0.16 ml/min and that of saliva while chewing six different foods was 3.6 +/- 0.8 ml/min. The mean times spent eating, and awake but not eating, were 80.8 +/- 27.3 and 820 +/- 59 min, respectively, and the volumes of saliva produced during those periods would average about 288 and 208 ml, respectively. If the flow rate is virtually zero during sleep, the estimated total salivary volume produced per day is calculated to be about 500 ml. PMID- 7487583 TI - Rifabutin use in inflammatory bowel disease. PMID- 7487582 TI - Vancomycin use in ophthalmology. PMID- 7487584 TI - Treatment of cytomegalovirus retinitis with intraocular sustained-release ganciclovir implant. PMID- 7487585 TI - Disease prevention and health promotion. PMID- 7487586 TI - The corneal endothelium after myopic excimer laser photorefractive keratectomy. PMID- 7487587 TI - Inefficacy of low-dose intraoperative fluorouracil in the treatment of primary pterygium. PMID- 7487588 TI - Detecting varicella-zoster virus DNA in iridocyclitis using polymerase chain reaction: a case of zoster sine herpete. PMID- 7487590 TI - Bilateral orbital varices associated with habitual bending. PMID- 7487589 TI - Aminoglycoside macular infarction in association with gentamicin-soaked collagen corneal shield. PMID- 7487592 TI - The use of silicone oil for HIV-related retinal detachment. PMID- 7487591 TI - Uterine cervical carcinoma metastasis to subconjunctival tissue. PMID- 7487593 TI - Controversies in endophthalmitis prevention. The risk for emerging resistance to vancomycin. PMID- 7487595 TI - Managed care in JAMA and the Archives journals. A call for papers for coordinated theme issues [ editorial ]. PMID- 7487594 TI - The ERG is alive and well. PMID- 7487596 TI - An updated standard for clinical electroretinography. AB - The value of the electroretinogram (ERG) in clinical practice depends in part on its consistency and the ability to compare results with other laboratories and the literature. These issues were addressed by the publication of the International Standard for Clinical Electroretinography in 1989. This Standard provided technical information and defined a core of basic test procedures and test responses (Figure) to ensure that ERGs performed worldwide would be equivalent for clinical and research purposes. Most major laboratories now subscribe to the tenets of the Standard, and it should be noted that the Standard does not limit laboratories from doing additional specialized tests (beyond the recommendations) for either clinical purposes or research. Since science and medicine are not static, the original Standard mandated revision every 4 years by the International Society for Clinical Electrophysiology of Vision. The first revision, titled "Standard for Clinical Electrophysiology (1994 Update)," has just been published, and it contains changes and new provisions that should be reviewed carefully by those who perform ERGs. The major changes and additions in the updated ERG Standard are as follows. PMID- 7487597 TI - Cigarette smoking and risk for progression of nuclear opacities. AB - OBJECTIVE: To examine the relationships between smoking and the 5-year incidence of new nuclear opacities and between smoking and the progression of nuclear opacities in a prospective study of a cohort of Chesapeake Bay watermen. METHODS: A total of 442 men (age, > or = 30 years in 1985) with paired, gradable lens photographs in at least one eye in both 1985 and 1990 were studied. Photographs were graded by two readers who used the grading scheme of the Wilmer Institute, Baltimore, Md, with severity ranging in decimal units between 0.0 and 4.0. Data on the smoking history of the subjects were collected by personal interviews that were conducted in 1985 and updated in 1990. RESULTS: The incidence and progression of opacities increased with age. A nonsignificant association was observed between smoking (for both current and ex-smokers) and the incidence of a nuclear opacity. The risk of progression of nuclear opacities of less than grade 3 at baseline to grade 3 or worse was 2.4-fold higher among current smokers in 1985, compared with that among ex-smokers and nonsmokers (95% confidence limits: 1.0, 6.0) after adjustment for age, baseline opacity status, and alcohol use. An 18% increased risk of progression was significantly associated with each pack year that a subject smoked between 1985 and 1990. CONCLUSION: These data confirm previous findings that smoking is associated with a nuclear opacity, particularly with progression to severe opacities. PMID- 7487598 TI - Delayed herpes zoster pseudodendrites. Polymerase chain reaction detection of viral DNA and a role for antiviral therapy. AB - BACKGROUND: The late-onset pseudodendrites, delayed corneal mucous plaques, of herpes zoster ophthalmicus are reported to be of mechanical or immune origin and to be worsened by antiviral therapy. OBJECTIVE: To study pseudodendrites to ascertain a viral presence in the lesions and their response to antiviral therapy. DESIGN: Prospective clinical study. SETTING: Outpatient and inpatient hospital-based corneal specialty referral practice; molecular virology laboratory. PATIENTS: Six patients, aged 33 to 89 years, four with delayed herpes zoster ophthalmicus pseudodendrites and two with herpes zoster ophthalmicus neurotrophic ulceration. One patient was immunosuppressed. MAIN OUTCOME MEASURES: Findings from clinical evaluation; polymerase chain reaction assays of lesions and tear film of six patients; polymerase chain reaction and light and electron microscopy of the corneal button from one patient; and the clinical response of four patients to various antiviral drugs. RESULTS: In contrast to reports in the current literature, delayed pseudodendrites may also be infectious, as they are positive for zoster DNA by polymerase chain reaction and appear responsive to certain antiviral therapy. The corneal button from an immunosuppressed patient had mature and immature viral particles in the basal cells within 2 weeks of transplantation. CONCLUSIONS: To our knowledge, this is the first report of viral DNA in delayed zoster pseudodendrites. Recurrent viral infection may play a role in this form of zoster keratopathy and warrant antiviral therapy. PMID- 7487599 TI - Retinal microaneurysm counts and 10-year progression of diabetic retinopathy. AB - OBJECTIVE: To determine the relationship of change in the number of retinal microaneurysms to the 10-year progression to significant retinopathy, proliferative retinopathy, and clinically significant macular edema. DESIGN: Population-based study of persons with younger- and older-onset diabetes with 10 years of follow-up. SETTING AND PATIENTS: Eleven-county area in southern Wisconsin, where 189 patients with diabetes who had only retinal microaneurysms in photographs at baseline participated in 4- and 10-year follow-up examinations. OUTCOME MEASURES: Ten-year incidence of moderate nonproliferative diabetic retinopathy or worse, proliferative retinopathy, or clinically significant macular edema as determined by masked grading of stereoscopic color fundus photographs of seven standard fields. RESULTS: The increase in the number of retinal microaneurysms and the ratio of the number of retinal microaneurysms at the 4-year follow-up to the number at baseline were positively associated with incidence of proliferative retinopathy or clinically significant macular edema at the 10-year follow-up. Proliferative retinopathy was approximately 4.6 times and clinically significant macular edema was approximately 9.1 times more likely to develop at 10-year follow-up in eyes in which the number of microaneurysms increased by 16 or more from baseline to the 4-year follow-up than in eyes with no increase. Proliferative retinopathy was 3.4 times and clinically significant macular edema was 6.7 times more likely to develop at 10-year follow-up in eyes that had ratios of 3 or greater of the number of retinal microaneurysms at the 4 year follow-up to the number at baseline than in eyes in which the ratios were smaller. These relationships remained after controlling for the level of glycosylated hemoglobin and type of diabetes. CONCLUSIONS: Microaneurysm counts using stereoscopic color fundus photographs are an early important measure of progression of retinopathy and may serve as a surrogate end point for severe change in some clinical trials. PMID- 7487600 TI - Imaging of chorioretinal anastomoses in vascularized retinal pigment epithelium detachments. AB - OBJECTIVE: To identify specific features of pigment epithelium detachments with limited hyperfluorescent lesions (hot spots). DESIGN: One hundred eighty-two consecutive patients (186 eyes) who had vascularized pigment epithelium detachments and recent onset of symptoms were examined with indocyanine green and fluorescein videoangiography using the scanning laser ophthalmoscope. The choroidal neovascularization complex and macular retinal vessels were studied. The natural history and the effect of laser treatment were evaluated. RESULTS: Fifty-four eyes had hot spots on indocyanine green angiography. In 50 of these 54 eyes, the video analysis showed an anastomosis of one or more retinal vessels, with the choroidal neovascularization within the hot spot. One or two retinal veins or arteries or both filled with both dyes and were seen to enter into the hot spot. Results of indocyanine green-guided photocoagulation of the hot spot in 28 eyes were disappointing. CONCLUSION: Continuous recording of the early phases of fluorescein and indocyanine green angiography allowed identification of chorioretinal anastomoses in vascularized pigment epithelium detachments with hot spots at an early exudative stage of age-related macular degeneration in 50 (26.8%) of 186 eyes. The poor outcome of laser photocoagulation could be related not only to the development of an overlying pigment epithelium detachment, but also to the retinal and choroidal vascularization of the lesion. PMID- 7487601 TI - Silicone oil in repair of retinal detachments caused by necrotizing retinitis in HIV infection. AB - OBJECTIVE: To evaluate the safety and efficacy of 1000- and 5000-centistoke silicone oil as retinal tamponades for the treatment of retinal detachments secondary to necrotizing retinitis in patients with human immunodeficiency virus (HIV) infection. DESIGN: A prospective observational study. SETTING: Community and university-based ophthalmology clinics. PATIENTS: Three hundred fifty patients with HIV infection, who had 407 eyes with retinal detachments secondary to necrotizing retinitis. INTERVENTION: Vitrectomy surgery for retinal detachment with 1000- or 5000-centistoke silicone oil as the retinal tamponade. OUTCOME MEASURES: Efficacy was measured both by anatomic success (defined as complete retinal attachment or macular attachment) and by visual acuity success (defined as preservation of visual acuity or ambulatory vision). Safety was determined by the rate of complications, including abnormal intraocular pressure and corneal and lens opacification. RESULTS: At the last follow-up examination, the retina was completely attached in 287 (73%) of 393 eyes, the macula was attached in 370 eyes (94%), 268 eyes (68%) had ambulatory vision, and visual acuity was preserved in 219 (56%) of 388 eyes. Corneal opacification, hypotony, and silicone oil emulsification were present in 4%, 2%, and 1% of eyes, respectively. One eye had elevated intraocular pressure. Of the 57 patients who had both eyes treated, 35 died, of whom four (11%) had nonambulatory vision in both eyes. Of the 293 patients who had one eye treated, 122 died, of whom 44 (36%) died with nonambulatory vision in the treated eye. The median time to cataract was 192 days; to nonambulatory vision, 474 days; and to death, 204 days. CONCLUSIONS: Silicone oil repair of retinal detachments in necrotizing retinitis is an efficacious and safe procedure that delays or prevents loss of vision in advanced HIV disease. PMID- 7487602 TI - Golf-related ocular injuries. AB - OBJECTIVE: To review golf-related ocular injuries, which account for 1.5% to 5.6% of all sports injuries. METHODS: During 8 years (1986 to 1994), a retrospective review of sports-related trauma was performed at the Medical College of Wisconsin, Milwaukee. Eight blunt ocular injuries (four ruptured globes and four globes without rupture) caused by golf-related activities were identified. The four ruptured globes caused by golf-related trauma accounted for 1.2% of all penetrating injuries and/or ruptured globes and 11.7% of sports-related injuries. RESULTS: A golf ball projectile was the mechanism of injury in six patients, while two patients were struck with a gold club. The four patients with ruptured globes had an initial visual acuity of light perception or worse, and three globes were subsequently enucleated, while one was prephthisic. In the four trauma cases without rupture, surgical intervention was required to achieve anatomic stability, with final visual acuities ranging from 20/25 to 20/40. CONCLUSIONS: The incidence of ocular injuries caused by golf-related trauma is low compared with that for other sports-related injuries. Although the prognosis for ruptured globes occurring in this setting remains extremely guarded, blunt trauma without rupture caused by a golf-related injury is associated with a more favorable visual and anatomic outcome. PMID- 7487604 TI - Radiotherapy for thyroid orbitopathy. Effects on extraocular muscle balance. AB - OBJECTIVE: To investigate the effects of radiotherapy on the restrictive ophthalmopathy and strabismus that occur with Graves' orbitopathy. DESIGN: A prospective study of patients with motility disturbances due to thyroid orbitopathy, followed by serial quantitative measurements of ocular restrictions and deviations, both before and at 1 and 6 months after treatment with high energy radiotherapy. SETTING: Urban subspecialty private practice. PATIENTS: Thirty-three consecutive patients with restricted extraocular movements and diplopia secondary to thyroid orbitopathy who were treated with radiotherapy were seen during an 8-year period. MEASURES: Maximal horizontal and vertical ductions, plus ocular deviations in primary and reading positions of gaze, were quantitatively evaluated by one clinical investigator to ensure consistency. RESULTS: Statistically significant improvement in gaze restriction on supraduction and improvement of vertical and horizontal strabismus were demonstrated at follow-up visits after treatment. CONCLUSIONS: Although statistically significant improvement of motility disturbances were demonstrated after radiotherapy, there was less than desirable functional improvement of restrictive ophthalmopathy and strabismus; thus the usefulness of radiotherapy was limited when it was used alone to treat diplopia. PMID- 7487603 TI - Abnormal extracellular material in the levator palpebrae superioris complex in congenital ptosis. AB - OBJECTIVE: To determine the pathologic abnormalities of the levator palpebrae superioris in congenital ptosis. METHODS: By means of light and electron microscopy and immunohistochemistry, anterior levator tissue specimen from 15 patients with congenital ptosis excised during routine levator resections were examined. RESULTS: All specimens showed lack of muscle fibers with endomysial and perimysial fibrosis and thickening of the aponeurosis. In addition, in four of the 15 patients, an abnormal extracellular material was present. By light microscopy this appeared as an unusual amorphous material arranged in clumps and bands, and electron microscopy showed it to consist of parallel coarse bundles of fibrillogranular material. Collagen type III and fibronectin were identified within this material by immunohistochemistry. There was no detectable collagen types I, II, IV, V, VI, or VII or laminin, and the material did not stain for actin, myosin, myoglobin, amyloid P component, or amyloid A. CONCLUSIONS: In four of 15 samples of levator palpebrae superioris from patients with congenital ptosis, we identified an unusual amorphous extracellular material that stained positively for collagen type III and fibronectin on immunohistochemistry. This novel material, which we call "amorphocollagenoid," may represent a product of dysgenesis of the levator tissues. The source and full composition of this material merit further study. PMID- 7487605 TI - Management of diplopia limited to down gaze. AB - OBJECTIVE: To evaluate the usefulness of various optical and surgical treatment modalities in the treatment of patients who were symptom free in the primary position of gaze yet had symptomatic diplopia in the reading position (down gaze at near). PATIENTS AND METHODS: A retrospective chart review was conducted to identify all patients with presbyopia I have treated who were symptom free in the primary position but had diplopia in down gaze associated with vertically incomitant strabismus. Of 51 patients identified, 32 were symptomatic in down gaze due to a hypertropia associated with a unilateral inferior rectus muscle underaction. Twenty-two of these patients had previously undergone recession of the affected inferior rectus muscle for treatment of thyroid eye disease; four patients, for correction of entrapment secondary to blow-out orbital fracture; and four patients, for treatment of superior oblique muscle palsy. Two patients had undergone prior surgery for superior oblique myokymia. Eight patients had horizontal diplopia associated with an A pattern, and eight patients had horizontal diplopia associated with a V pattern. Three patients had vertical and horizontal diplopia after partial recovery from third-nerve palsy. RESULTS: The treatment modalities were varied and individualized. Treatment consisted of optical management (20 patients), surgical management (21 patients), or a combination of both (10 patients). These treatment modalities resulted in comfortable single binocular vision for reading in 41 of the 51 patients. Successful optical treatment consisted of Fresnel prisms (four patients), slab off prisms (two patients), single-vision readers (seven patients), switch to nonprogressive bifocal lenses (three patients), and a high bifocal segment (16 patients). Successful surgical modalities included posterior fixation of the contralateral inferior rectus muscle (10 patients), surgery for A- or V- pattern strabismus (five patients), or bilateral inferior rectus muscle recession (six patients). CONCLUSION: Diplopia in the reading position frequently can be alleviated with a systematic approach that includes both optical and surgical modalities. PMID- 7487606 TI - Dystrophic calcification of an implanted hydroxyethylmethacrylate intraocular lens. AB - Hydroxyethylmethacrylate is a biomaterial still under clinical trial for use in foldable intraocular lenses. We observed a patient in whom a geographic opacification developed within an implanted hydroxyethylmethacrylate lens, together with granular deposits on the posterior lens capsule and in the scar of a paracentesis. The intraocular lens and posterior lens capsule were removed because of impaired visual acuity. Light and scanning electron microscopy disclosed nodular calcifications within the intraocular lens and granular, partially crystalline, calcifications on the posterior lens capsule. Energy dispersive x-ray analysis and x-ray diffraction showed the deposits in the intraocular lens to consist of calcium hydroxyapatite. We presume this mineralization to be dystrophic, with calcium derived from lens remnants and phosphorus possibly derived from a thymoxamine solution used briefly during the cataract operation. Our observation suggests caution in the use of phosphated solutions together with hydroxyethylmethacrylate intraocular lenses and may warrant reconsideration of the suitability of hydroxyethylmethacrylate intraocular lenses, should additional similar cases be reported. PMID- 7487607 TI - Retinal pigment epithelial wound healing in vivo. AB - OBJECTIVE: To develop an in vivo rabbit model of retinal pigment epithelial wound healing that preserves the overlying retina. METHODS: Hydraulic debridement of the retinal pigment epithelium was performed in one eye of 35 pigmented rabbits by means of a pars plana vitrectomy approach. Five of the 35 eyes were examined by stereoscopic color fundus photography, fluorescein angiography, and light microscopy on each of the following postoperative days: 0, 2, 4, 7, 14, 28, and 56. RESULTS: Retinal pigment epithelial debridement with this technique results in apical decapitation of the retinal pigment epithelial cells followed by subsequent hydraulic removal of the residual nucleus-containing basal cellular debris. The retinal pigment epithelium-denuded Bruch's membrane was resurfaced mostly by a monolayer of flattened, hypopigmented retinal pigment epithelial cells within 4 days after debridement. Progressive retinal pigment epithelial hyperplasia also occurred beginning between postoperative days 2 and 4. CONCLUSIONS: Retinal pigment epithelial wound healing after hydraulic debridement occurs rapidly and in a manner initially consistent with sliding migration. Progressive retinal pigment epithelial hyperplasia also occurs and may contribute to this repair process. Further investigation of retinal pigment epithelial repair by means of this in vivo model may provide important insight into the pathogenesis and treatment of outer retinal disorders. PMID- 7487609 TI - Stickler syndrome. A mutation in the nonhelical 3' end of type II procollagen gene. AB - BACKGROUND: All of the mutations in the type II procollagen (COL2A1) gene that have been identified in families affected with Stickler syndrome have been located primarily in the triple helical region of the gene. We report what we believe is the first premature stop codon in the globular C-propeptide region encoded by the COL2A1 gene, in a family affected with Stickler syndrome. DESIGN: Genomic DNA from affected and unaffected family members of this three-generation family was amplified using the polymerase chain reaction. The polymerase chain reaction products were directly sequenced for DNA analysis. RESULTS: Direct sequencing showed a single base deletion in exon 50, resulting in a premature stop codon in exon 51 in the globular C-propeptide of COL2A1 gene in all affected members. CONCLUSIONS: These results implicate premature stop codons as a common cause of Stickler syndrome. The location of this premature stop codon in the far end of the nonhelical 3' end of the gene indicates that a truncated C-propeptide of at least 84 amino acid residues is inadequate for the functional gene product. PMID- 7487608 TI - Clinically successful contact transscleral krypton laser cyclophotocoagulation. Long-term histopathologic and immunohistochemical autopsy findings. AB - OBJECTIVE: To report long-term histopathologic findings 10 months after contact transscleral krypton cyclophotocoagulation. METHODS: The tissue response in a successfully treated eye was analyzed by light microscopy and a panel of 11 antibodies to epithelial, mesenchymal, and inflammatory cells. RESULTS: A 75-year old man with uncontrolled angle recession glaucoma was treated with transscleral contact krypton cyclophotocoagulation (17 burns, 3.5 J each) 10 months before his death. The intraocular pressure fell from 28 to 17 mm Hg 6 months after therapy. Confluent scars straddled the posterior pars plicata and the anterior pars plana. The ciliary processes were destroyed, but the sclera and zonules were intact. Vimentin and cytokeratin 8 and 18 persisted in the degenerated ciliary epithelium. The inner connective-tissue layer and the ciliary muscle had atrophied, as shown with antibodies to the HNK-1 epitope, desmin, and alpha smooth-muscle actin. Macrophages with phagocytized pigment and single T cells were present instead. No unusual inflammatory infiltrate was present in the choroid of either eye. CONCLUSIONS: Clinically effective ablation of ciliary processes is achieved with contact krypton laser. Little chronic inflammation and no signs of sympathetic ophthalmia were present. Atrophy of the ciliary muscle may reduce accommodative capacity in younger patients undergoing cyclophotocoagulation. PMID- 7487611 TI - Double-mirror peripheral vitrectomy lens. AB - Many surgeons use prism lenses to see the periphery of the fundus during vitrectomy; however, chromatic aberrations in higher-power prismatic lenses cause blurring of the peripheral image. For better visualization of the periphery of the fundus, we developed a new contact lens, the double-mirror peripheral vitrectomy lens. The new lens is a quartz cylinder with two mirrors, and it provides a crisp, clear, upright image of much more of the peripheral fundus than is visible through conventional prism lenses. The new lens also provides a wider area of view than conventional prism lenses. PMID- 7487612 TI - Ultrasound biomicroscopy in the management of malignant melanoma of the iris. PMID- 7487610 TI - Wedge resection of eyelid margin in the treatment of abnormal eyelid margins. AB - Widened, hypertrophic, erythematous eyelid margins occasionally occur secondary to congenital anomalies or previous surgical procedures. When a full-thickness resection of the eyelid is not possible because of horizontal tightness of the lid, it is possible to excise a wedge of the eyelid margin itself. This procedure was successful in decreasing widened, hypertrophic, erythematous eyelid margins in three patients. PMID- 7487613 TI - Calcific retinal emboli and collateral shunting in a woman with rheumatic heart disease. PMID- 7487614 TI - Results of the Endophthalmitis Vitrectomy Study. A randomized trial of immediate vitrectomy and of intravenous antibiotics for the treatment of postoperative bacterial endophthalmitis. Endophthalmitis Vitrectomy Study Group. AB - OBJECTIVE: To determine the roles of immediate pars plana vitrectomy (VIT) and systemic antibiotic treatment in the management of postoperative endophthalmitis. DESIGN: Investigator-initiated, multicenter, randomized clinical trial. SETTING: Private and university-based retina-vitreous practices. PATIENTS: A total of 420 patients who had clinical evidence of endophthalmitis within 6 weeks after cataract surgery or secondary intraocular lens implantation. INTERVENTIONS: Random assignment according to a 2 x 2 factorial design to treatment with VIT or vitreous tap or biopsy (TAP) and to treatment with or without systemic antibiotics (ceftazidime and amikacin). MAIN OUTCOME MEASURES: A 9-month evaluation of visual acuity assessed by an Early Treatment Diabetic Retinopathy Study acuity chart and media clarity assessed both clinically and photographically. RESULTS: There was no difference in final visual acuity or media clarity with or without the use of systemic antibiotics. In patients whose initial visual acuity was hand motions or better, there was no difference in visual outcome whether or not an immediate VIT was performed. However, in the subgroup of patients with initial light perception-only vision, VIT produced a threefold increase in the frequency of achieving 20/40 or better acuity (33% vs 11%), approximately a twofold chance of achieving 20/100 or better acuity (56% vs 30%), and a 50% decrease in the frequency of severe visual loss (20% vs 47%) over TAP. In this group of patients, the difference between VIT and TAP was statistically significant (P < .001, log rank test for cumulative visual acuity scores) over the entire range of vision. CONCLUSIONS: Omission of systemic antibiotic treatment can reduce toxic effects, costs, and length of hospital stay. Routine immediate VIT is not necessary in patients with better than light perception vision at presentation but is of substantial benefit for those who have light perception-only vision. PMID- 7487615 TI - Primary corneal graft failure. A national reporting system. Medical Advisory Board of the Eye Bank Association of America. AB - OBJECTIVES: To describe a national eye banking registry and to assess the effects of donor age, cause of donor death, time from death to procurement, storage time, and distance between the points of recovery and transplantation on the reported occurrence of primary corneal graft failure. DESIGN: We performed a retrospective case-control study to estimate the odds ratios of five donor factors for cases of primary graft failure voluntarily reported to a national registry using controls from selected eye banks. We also performed a nested case-control cohort study to compare cases of primary graft failure that occurred in both corneas from the same donor with those of nonmated corneas in which primary graft failure was reported to assess odds ratios for the same donor factors. PATIENTS: One hundred forty-seven patients developed primary graft failure in penetrating keratoplasty transplantations performed between January 1, 1991, and December 31, 1993. These cases were reported to the Adverse Reaction Registry of the Eye Bank Association of America, Washington, DC. Controls included 7240 donor corneas distributed by nine eye banks during 1992. RESULTS: Of the 147 donor corneas that developed primary graft failure, 17 (12%) were obtained from donors who were aged 70 years and older, 39 (27%) came from donors who died of trauma, 13 (9%) had a cadaver time longer than 12 hours, 10 (7%) had a storage time longer than 7 days, and 38 (26%) were distributed outside the eye bank's region. Compared with controls, these donor corneas were more likely to have a storage time longer than 7 days (odds ratio, 2.4; 95% confidence limits, 1.2 and 4.6) and to come from donors aged 70 years and older (odds ratio, 2.4; 95% confidence limits, 1.4 and 4.0). The 22 corneas (15%) in which primary graft failure occurred in both recipients from the same donor were 8.5 times (95% confidence limits, 1.1 and 51.5) more likely to be preserved beyond 1 week than were nonmated corneas with primary failure but were not from significantly older donors. Logistic regression analysis showed that the association between prolonged storage time and primary graft failure in mated corneas remained significant even when the analysis was controlled for other donor factors. CONCLUSIONS: No clearly defined donor or eye banking factor accounted for most cases of primary graft failure, although prolonged corneal storage and advanced donor age may increase its risk. Ophthalmologists are urged to report to their eye bank all cases of primary graft failure and other adverse events that might be attributable to donor eye tissue. PMID- 7487616 TI - Outcome of penetrating keratoplasty after ocular trauma in children. AB - OBJECTIVE: To characterize the outcome of penetrating keratoplasty in children for the visual rehabilitation of corneal injury. DESIGN: Multicenter retrospective study of the outcome of penetrating keratoplasty for ocular trauma in children. PATIENTS: All children aged 12 years and younger who underwent penetrating keratoplasty between January 1975 and May 1993 for penetrating corneal (n = 18) or corneoscleral (n = 3) trauma and nonpenetrating corneal trauma (n = 1). MAIN OUTCOME MEASURES: Graft survival and postoperative visual acuity improvement. RESULTS: Twenty-two children underwent 25 penetrating keratoplasties during the study period. Graft survival was 84% at 1 year and 70% at 2 years after surgery. Visual acuity improved in 15 (83%) of the 18 children with measurable preoperative and postoperative vision. Children with posterior segment injury before keratoplasty were less likely to maintain a clear graft (P = .04) and less likely to have improved vision postoperatively (P = .06). Children who received amblyopia treatment and appropriate optical correction postoperatively were more likely to have visual improvement than those who did not (P = .02). CONCLUSIONS: Penetrating keratoplasty for corneal trauma is generally a successful operation in the pediatric age group, with visual improvement seen in more than four fifths of cases. Graft survival and visual outcome are best in patients with trauma limited to the anterior segment. Aggressive postoperative refractive correction and amblyopia management also appear to contribute to a better visual outcome in pediatric keratoplasty. PMID- 7487617 TI - Reproducibility and responsiveness of the VF-14. An index of functional impairment in patients with cataracts. AB - OBJECTIVES: To assess the test-retest reliability and responsiveness of the VF 14, which is an index of functional impairment in patient with cataracts. DESIGN: Observational longitudinal study. Patients were enrolled prior to undergoing their first cataract surgery between July 15 and December 15, 1991, and they were followed up for 1 year after surgery. SETTING: Patients were recruited from 72 ophthalmologists' practices in three US cities. PATIENTS: Five hundred fifty-two patients who had undergone a surgical procedure in only one eye by the 4-month postoperative follow-up (responsiveness analyses) and a subset of these (n = 426) who had not subsequently undergone surgery for the second eye by the 12-month postoperative follow-up (reproducibility analyses). MAIN OUTCOME MEASURES: Two health status measures (the VF-14 and the Sickness Impact Profile, two global measures of a patient's trouble and satisfaction with his or her vision, and best corrected visual acuity in each eye. RESULTS: The VF-14 is highly reproducible, with an intraclass correlation coefficient of .79 when patient-rated criteria are used to define stable patients. The intraclass correlation coefficient was lower (.57 to .71) when various measures of visual acuity were used to define stable patients. The VF-14 is also about three times more responsive to a change in vision than the Sickness Impact Profile, which is a generic health status measure (effect size of approximately 1.00 vs 0.30). Estimates of the responsiveness of the VF-14 and the Sickness Impact Profile were not associated with preoperative visual acuity in the operated on or better eye. Responsiveness of the VF-14, however, was higher in patients with greater self-rated trouble with vision preoperatively. CONCLUSIONS: The VF-14 was reproducible in stable patients during an 8-month period, and it was more responsive to clinically significant changes in vision than was a generic health status measure (ie, the Sickness Impact Profile). PMID- 7487618 TI - Effects of brimonidine on aqueous humor dynamics in human eyes. AB - OBJECTIVE: To evaluate the mechanism by which brimonidine, a selective alpha 2 adrenergic agonist, lowers intraocular pressure (IOP) in humans. SUBJECTS: Twenty one volunteers with ocular hypertension. METHODS: Brimonidine tartrate (0.2%) was given topically twice daily for 1 week to one eye in a randomized, double-masked study. The fellow eye was similarly treated with brimonidine vehicle. Before (baseline) and after 1 week (day 8) of dosing, IOP, aqueous flow, episcleral venous pressure, and tonographic outflow facility were directly measured. Fluorophotometric outflow facility and uveoscleral outflow were calculated. Brimonidine-treated eyes were compared with vehicle-treated contralateral control eyes and with baseline measurements after 1 week of dosing. RESULTS: Brimonidine significantly (P < .001, Student's two-tailed t test) reduced IOP mean +/- SE of 4.7 +/- 0.7 and 4.2 +/- 0.4 mm Hg compared with the baseline day and with the vehicle-treated contralateral control eyes, respectively. Compared with the baseline day, aqueous flow was reduced by 20% (P = .002) and uveoscleral outflow was increased (P = .04). A slight contralateral decrease in IOP of 1.2 +/- 0.6 mm Hg (P = .05) and in aqueous flow of 12% (P = .05) was noted. No significant difference was seen in the outflow facility values or episcleral venous pressure compared with the baseline day or with the contralateral control eye. CONCLUSIONS: The brimonidine-induced reduction in IOP in humans is associated with a decrease in aqueous flow and an increase in uveoscleral outflow. The decrease in IOP and aqueous flow in the contralateral control eye on day 8 compared with the baseline day suggests a mild contralateral effect. PMID- 7487619 TI - Serum antioxidants and age-related macular degeneration in a population-based case-control study. AB - OBJECTIVE: To investigate relationships between levels of tocopherols and carotenoids in the serum and age-related macular degeneration (ARMD). DESIGN: A nested case-control study within a population-based cohort. PARTICIPANTS: Cases included a sample of subjects with retinal pigment abnormalities with the presence of soft drusen (n = 127) or with late ARMD (geographic atrophy [n = 9]) or neovascular and exudative macular degeneration (n = 31). An equal number of controls (167 pairs) were selected from among participants in the Beaver Dam Eye Study. The controls had no photographic evidence of soft drusen, retinal pigment abnormalities, or late ARMD and were matched with cases for age, sex, and current smoking status. DATA COLLECTION: Presence and severity of ARMD were determined from masked grading of fundus photographs obtained from 1988 to 1990. Levels of individual carotenoids and tocopherols were determined in serum collected at the same time. RESULTS: Average levels of individual carotenoids were similar in cases and controls. Average levels of vitamin E (alpha-tocopherol) were lower in people with exudative macular degeneration (P = .03). However, the difference was no longer statistically significant after controlling for levels of cholesterol in the serum. Persons with levels of lycopene, the most abundant carotenoid in the serum, in the lowest quintile were twice as likely to have ARMD. Levels of the carotenoids that compose macular pigment (lutein with zeaxanthin) in the serum were unrelated to ARMD. CONCLUSIONS: Very low levels of one (lycopene) but not other dietary carotenoids or tocopherols were related to ARMD. Lower levels of vitamin E in subjects with exudative macular degeneration compared with controls may be explained by lower levels of serum lipids. PMID- 7487620 TI - Presumed acquired ocular toxoplasmosis. AB - OBJECTIVE: To describe the clinical characteristics and laboratory findings of eight patients with focal chorioretinitis presumably caused by acquired toxoplasmosis. DESIGN: Case series. SETTING: Referral hospitals in the Netherlands. PATIENTS: Eight patients, aged 42 to 75 years, with unilateral focal chorioretinitis and laboratory evidence of a recently acquired infection with Toxoplasma gondii. MAIN OUTCOME MEASURES: Findings from ocular examination and analysis of both serum and aqueous humor samples for Toxoplasma and viral antibodies. RESULTS: All patients had unilateral focal chorioretinitis without associated old scars in the posterior pole. Patients treated with systemic or periocular corticosteroids not accompanied by antiparasitic medication showed a rapid increase of inflammation. All eight patients had Toxoplasma IgM antibodies in their serum samples, seven of whom had high Toxoplasma IgG titers. Five of eight patients had increased intraocular production of IgG antibodies against T gondii. CONCLUSION: Unilateral focal chorioretinitis in patients of any age should alert the clinician to consider acquired ocular toxoplasmosis in the differential diagnosis. PMID- 7487621 TI - Partly accommodative esotropia. Should you overcorrect and cut the plus? AB - OBJECTIVES: To investigate the long-term motor stability and sensory outcome of patients with partly accommodative esotropia who were overcorrected surgically and in whom the hyperopic correction was reduced postoperatively, and to determine if those results depended on the amount of hyperopia present. DESIGN: A 15-year prospective study that analyzed 5-year outcome. Patients whose esotropia was not initially overcorrected were used as controls. PATIENTS: Of 382 patients who underwent surgery for partly accommodative esotropia, 22 were surgically overcorrected and were followed up for 5 years. RESULTS: Of the eight patients in the study group with 2.5 diopters or less of hyperopia in their fixing eye, seven had good motor alignment compared with four of 14 patients who had more than 2.5 diopters of hyperopia. Ninety-one percent (148/163) of the control patients who had greater than 2.5 diopters of hyperopia maintained good motor alignment 5 years after surgery compared with 29% of the study group patients. This difference was statistically significant. Of the eight study patients with less than 2.5 diopters of hyperopia, five developed good stereopsis compared with one of 14 patients with greater hyperopia. CONCLUSIONS: Surgical overcorrection in patients with partly accommodative esotropia with greater than 2.5 diopters of hyperopia may not be reversible by postoperative reduction in the hyperopic correction. It often is reversible, however, in patients with 2.5 diopters or less of hyperopia. PMID- 7487622 TI - Early progression of ophthalmoplegia in patients with ischemic oculomotor nerve palsies. AB - OBJECTIVE: To describe the frequency and clinical correlates of early progression of ophthalmoplegia in patients with ischemic oculomotor nerve palsies. DESIGN: Cohort survey, case series. SETTING: Multispecialty clinic providing primary, secondary, and tertiary care in central and northern Wisconsin. PATIENTS: Sixteen patients evaluated within 1 week of the reported onset of ischemic oculomotor nerve palsy were identified and followed up prospectively using a standardized ophthalmoplegia grading scheme. All patients were followed up serially until their ophthalmoplegia resolved. MAIN OUTCOME MEASURES: Descriptive analysis of the temporal course of ophthalmoplegia and frequency of progression of deficits. Comparison between the group that had progression of ophthalmoplegia with the group that did not for age, hematocrit, cholesterol level, and adiposity; presence of diabetes, hypertension, hypercholesterolemia, and coronary artery disease; history of stroke; and tobacco use. RESULTS: Eleven (69%) of 16 patients had progression of ophthalmoplegia. The median time between reported onset and peak severity of ophthalmoplegia was 10 days. The only important difference between the progressive and nonprogressive groups was a shorter time to resolution of ophthalmoplegia for the nonprogressive group. CONCLUSIONS: Early progression of ophthalmoplegia occurs often in patients with ischemic oculomotor nerve palsies. The power to find differences between progressive and nonprogressive groups was limited by the small number of patients available for analysis. PMID- 7487623 TI - Hypoxic regulation of vascular endothelial growth factor in retinal cells. AB - BACKGROUND: Vascular endothelial growth factor (VEGF) is an angiogenic protein and vasopermeability factor whose intraocular concentrations are closely correlated with active neovascularization in patients with diabetes mellitus, central retinal vein occlusion, retinopathy of prematurity, and rubeosis iridis. OBJECTIVE: To determine whether hypoxia could induce expression of VEGF in retinal cells, which then promotes retinal endothelial cell proliferation. METHODS: Retinal pigment epithelial cells, pericytes, and microvascular endothelial cells were exposed to hypoxic conditions in vitro, and RNA expression of VEGF was evaluated by Northern blot analysis. The VEGF-specific proliferative potential of the medium was measured by means of retinal endothelial cell growth assays and VEGF-neutralizing VEGF receptor IgG chimeric protein. RESULTS: The VEGF RNA levels increased within 4 hours and reached elevations of threefold to 30-fold after 18 hours of hypoxia (0% to 5% oxygen, 5% carbon dioxide, 90% to 95% nitrogen) in all cell types (.01 < P < .03). Stimulation was dependent on oxygen concentration. The VEGF RNA levels were normalized by reinstitution of normoxia for 24 hours (P < .004). Medium conditioned by hypoxic retinal pericytes and retinal pigment epithelial cells stimulated retinal endothelial cell growth by 20% (P = .04), and this stimulation was entirely inhibited by VEGF-neutralizing receptor chimeric protein (P = .02). CONCLUSION: Hypoxia increases VEGF expression in retinal cells, which promotes retinal endothelial cell proliferation, suggesting that VEGF plays a major role in mediating intraocular neovascularization resulting from ischemic retinal diseases. PMID- 7487624 TI - Pathologic human vitreous promotes contraction by fibroblasts. Implications for proliferative vitreoretinopathy. AB - OBJECTIVE: To establish and quantify the presence of contraction-stimulating activity in pathologic vitreous and correlate this activity with clinical presentation and outcome, especially with proliferative vitreoretinopathy. METHODS: Contraction-stimulating activity of vitreous collected during surgery was quantified with a tissue culture assay using fibroblasts as target cells. The activity of each sample was correlated with patient history, clinical presentation, risk factors, proliferative disease, and postoperative proliferation. RESULTS: Pathologic vitreous contained measurable quantities of contraction-stimulating activity and stimulated contraction in vitro, with elevated activities in samples from patients with proliferative vitreoretinopathy, epimacular proliferation, retinal detachment, retinal defects, pigmented cells in the vitreous, hemorrhage, or uveitis. Patients with postoperative proliferation had significantly elevated mean activities. CONCLUSIONS: Levels of contraction-stimulating activity in pathologic vitreous correlate with some risk factors for the development of proliferative vitreoretinopathy and may ultimately be useful in the assessment of disease severity and the prediction of postoperative proliferation. PMID- 7487625 TI - Primary corneal graft failure. A national reporting system. PMID- 7487626 TI - The endophthalmitis vitrectomy study. PMID- 7487627 TI - Submacular surgery. Are randomized trials necessary? PMID- 7487628 TI - Histopathologic study of autosomal dominant vitreoretinochoroidopathy in a 26 year-old woman. AB - The clinicopathologic findings were obtained from enucleated eyes, obtained post mortem, of a 26-year-old woman with autosomal dominant vitreoretinochoroidopathy. Light microscopy demonstrated atrophic, disorganized peripheral retina with retinal blood vessels obscured by pigmented cells surrounding periodic acid Schiff-positive deposits. Peripheral retinal pigment epithelial cells showed multilayering and pigmentation, with a thickened basal lamina. By electron microscopy, the peripheral retinal vessel endothelium was replaced by an arrangement of morphologically polarized pigmented cells of presumed retinal pigment epithelial origin oriented with their basal surfaces toward a fibrillar matrix occupying the vessel lumen. The similarity of the findings in this young patient to those of an aged patient described previously suggest that autosomal dominant vitreoretinochoroidopathy is an early-onset dystrophy of the peripheral retina with minimal subsequent progression, characterized by a retinal pigment epithelial response that includes marked intraretinal migration and extracellular matrix deposition. PMID- 7487629 TI - Anterior segment trauma and air bags. PMID- 7487630 TI - A case of traumatic lens extraction. PMID- 7487631 TI - The use of perfluorocarbon liquid in the removal of radiopaque intraocular glass. PMID- 7487632 TI - Choroidopathy in multiple evanescent white dot syndrome. PMID- 7487633 TI - Orbital involvement of cervicofacial necrotizing fasciitis. PMID- 7487634 TI - A new device for noncontact wide-angle viewing of the fundus during vitrectomy. AB - Noncontact wide-angle viewing of the fundus during vitrectomy has heretofore relied on an image inverter and a 90-diopter lens fixed with a tube under the microscope. A new device in which the 90-diopter lens is not connected to the microscope but, rather, is connected to an independent support system is described. This device offers many advantages: (1) focusing on the various parts of the eye during vitreous surgery need not be done manually but is achieved by the up or down movement of the microscope, (2) translation of the microscope is easier during tilting of the eye, (3) a lateral slit-lamp can be easily added for illumination of the fundus, and (4) there is no risk that the 90-diopter lens will touch the cornea. PMID- 7487636 TI - Necrotic choroidal melanoma with orbital inflammation. PMID- 7487637 TI - Misuse of nonophthalmic and ophthalmic drops due to packaging similarity. PMID- 7487635 TI - Pigment dispersion secondary to cysts of the iris pigment epithelium. PMID- 7487638 TI - High incidence of topical allergic reactions to 1% apraclonidine. PMID- 7487639 TI - Tapered orbital retractors. PMID- 7487641 TI - Books, books, books. PMID- 7487640 TI - Paraneoplastic cone dysfunction: an unusual visual remote effect of cancer. PMID- 7487642 TI - Audiological assessment of infants and children with preauricular tags. AB - A previous study found sensorineural hearing loss in 13% of 178 children with solitary preauricular tags. The present results are quite different. Audiological assessment was performed on 42 children (median age of 2 months) with preauricular tags as the sole anatomical malformation. Tests included behavioral audiometry and transient evoked otoacoustic emissions. Twenty-two subjects were also tested with auditory brainstem recording while under general anesthesia during surgical excision of the tag. None of the children had a sensorineural hearing loss, but 4 had a bilateral or unilateral conductive hearing loss due to secretory otitis media. It is concluded that solitary ear tags are not a high risk factor for sensorineural hearing loss if not associated with a family history of hearing loss. PMID- 7487645 TI - The effect of broad-band noise on the binaural interaction components of human auditory brainstem-evoked potentials. AB - Three-channel Lissajous trajectories (3-CLTs) of binaural interaction components (BI) of auditory brainstem potentials (ABEPs) were derived from 13 normally hearing adults by subtracting the response to binaural clicks from the algebraic sum of monaurally evoked responses to clicks. ABEPs were recorded in response to 65 dB nHL, alternating-polarity clicks, presented at a rate of 11/s. The procedure was repeated with clicks alone as well as with clicks with broad-band masking noise. Noise was presented at 25 and 45 dB nHL, producing a signal-to noise ratio of +40 and +20 dB, respectively. All BI 3-CLTs included 6 planar segments (labeled BdI, BdII, BdIII, BeI, BeII and Bf) whose apex latencies, except Bf, increased with increasing noise level above 25 dB nHL, and whose durations, sizes, shapes and orientations did not change across noise levels. There were also significant increases in peak latencies of the BI from single channels vertex-mastoid and vertex-neck with increasing noise level. No significant change was found in the trajectory amplitude of apices, with the exception of apices BdIII and Bf whose amplitudes increased with increasing noise level. We suggest that the paradoxical increase in BI amplitude with masking noise may reflect a binaural enhancement of the effect of noise. The effects observed indicate that, whereas the response to clicks displays occlusion, the response to noise displays spatial facilitation at the brainstem level. PMID- 7487643 TI - Tympanogram recorded under cervical compression. AB - Tympanograms were recorded during compression of cervical veins in 19 normal ears to investigate whether pressure elevation in the middle and inner ears affects acoustic impedance. Upon cervical compression, the peak pressure of the tympanogram shifted to the positive pressure side. The peak amplitude of the tympanogram also changed in a manner similar to that after stapedius muscle contraction. These results suggest that the impedance changes caused by cervical compression reflected changes in both perilymphatic pressure and middle ear pressure. PMID- 7487644 TI - Effects of click polarity on brainstem auditory-evoked potentials in cochlear hearing loss: a working hypothesis. AB - The rarefaction-condensation differential potential (RCDP) obtained by subtracting brainstem auditory-evoked potentials (BAEPs) to C clicks from those to R clicks has been studied in 32 normal subjects and 31 cases of cochlear hearing loss. In normal subjects, no RCDP was recorded along the lower 30-55 dB of the JV latency-intensity function, thus defining the pre-RCDP range. The pre RCDP range was always abolished in losses unmasking BAEPs from lower ( < 1 kHz) tonotopic regions. When the BAEP originated from higher ( > 1 kHz) tonotopic regions, the pre-RCDP range was either reduced or abolished. These results led to a working hypothesis based on single-unit data and stating a dual dependence of polarity effects on variables distributed along the tonotopic and intensity dimensions, with respective break-points at 1 kHz, and at the junction of the tip and tail of unit frequency tuning curves. The 1 kHz break-point could represent the upper frequency limit for phase locking in man. PMID- 7487647 TI - Tympanometric screening in elementary school children. AB - The revised ASHA (American Speech Language Hearing Association, 1990) hearing screening guidelines were developed to decrease over-referrals for middle ear disorders through amended tympanometric measurements. These guidelines also recommended a large-scale normative study be done with tympanometric measures to further reduce the over-referral rate. Seven hundred and fifty ears from 392 kindergarten through third-grade children were tested. Normative values of Peak Ytm and tympanometric width were calculated by grade. For increasing grade level, there was an orderly increase in the mean Peak Ytm and a decrease in the mean tympanometric width. Sensitivity and specificity were calculated at 65 and 80% respectively, with a prevalence of 11%. Suggestions for improving sensitivity and specificity are discussed. PMID- 7487646 TI - Low-frequency biasing of round window responses in guinea pigs and chinchillas. AB - The acoustic biasing technique using low-frequency sound is of increasing interest to investigators, not only as a means of studying cochlear transduction but also as a promising tool for assessing cochlear pathology such as endolymphatic hydrops. We compared normal modulation patterns of round window responses in guinea pigs and chinchillas, whose low-frequency auditory characteristics are known to be different. A 50-Hz sine wave (90 dB SPL for guinea pigs and 80 dB SPL for chinchillas), which evoked an equivalent magnitude of cochlear microphonics (CM) in both species, was used to modulate the compound action potential (CAP) and the summating potential (SP) elicited by 8-kHz tone bursts. Overall patterns of CAP and SP modulation were almost identical between the two species except for a difference in the phase of 50 Hz CM. The phase of maximum SP enlargement was in accord with that of maximum CAP suppression, which led to inferred basilar membrane (BM) position at maximum scala tympani displacement by between 45 and 90 degrees. More complex or hysteresis effects seemed to be involved in the modulation of CAP and SP magnitude, in addition to the biasing effect owing solely to BM displacement. PMID- 7487648 TI - Case studies in lipid management. AB - Seven cases are described that highlight practical aspects of lipid management in Australian general practice. The major issues are strategies for obtaining reliable lipid values, interpreting the lipids in the context of overall cardiovascular risk, and appropriate use of pharmacotherapy in high risk patients. PMID- 7487649 TI - Sugar and fat, a recipe for disaster. New guidelines for the pharmacological management of diabetic dyslipidaemia. AB - Australia is embarking upon a more sophisticated approach to lipid-lowering, which is of particular relevance to those with diabetes. Diabetic dyslipidaemia is common and is usually managed in general practice. A step approach focusing on case finding, confirmation of diagnosis, lifestyle modification and pharmacological intervention should be cost effective in this high risk population (Figure 4). Although the focus of this review has been on diabetic dyslipidaemia, management should always consider overall cardiovascular risk. PMID- 7487650 TI - A review of reflex sympathetic dystrophy. AB - Reflex sympathetic dystrophy, or as it is now known, complex regional pain syndrome, Type 1, is an unusual complication of a variety of injuries whose development should be suspected in the presence of disproportionate pain coupled with autonomic disturbance. Early recognition and treatment with multimodality therapy offer a high probability of recovery. PMID- 7487651 TI - Endovascular surgery--an overview. AB - Endovascular surgery is the vascular equivalent of minimally invasive surgery. It offers the benefits of minimal morbidity and mortality rates as well as short hospital stay, with its associated cost curtailment. In spite of many technological innovations, only balloon dilatation and intravascular stenting have established their places in vascular surgery with the newer field of stent grafting for both occlusive and aneurysmal disease still being evaluated. PMID- 7487652 TI - The anaesthetic foot. A case of severe diabetic peripheral neuropathy. AB - Diabetic foot care is an important aspect of management. Self-care by the patient is of paramount importance. The case history described in this article highlights the absence of pain associated with peripheral neuropathy and the consequences of inadequate foot care by the patient. PMID- 7487653 TI - Critical thinking within the RACGP Training Program and the college examination. AB - Current developments with the formalisation of the Program's curriculum provide the opportunity for the College to clarify its position on this important educational area. While not all GPs want to be involved in research, increasing demands for accountability for expenditure of insurer's (ie the government's) funds are inevitable. Skills related to audit of actual practice process and outcomes are likely to be required in the near future. The option of developing an audit project segment for the College examination should be considered. Critical appraisal skills could be assessed in the context of the development of standards for the audit. Similarly the efficacy of the Training Program in the development of accurate self-assessment skills in trainees should be researched. If found lacking, steps should be put in place to ensure that these educational processes become effective. PMID- 7487654 TI - Alcohol and the nervous system. AB - Alcoholism is regarded as a multifactorial disorder, but the causes of alcoholism are poorly understood. The most important part of dealing with this widespread health and social problem is a prevention and early identification and treatment of those at risk. Therapeutic intervention should take place before development of cognitive impairment and physical dependence, otherwise treatment of developed alcoholism is notoriously unsatisfactory. PMID- 7487656 TI - Patient education. Benign positional vertigo. PMID- 7487655 TI - Treatment of Parkinson's disease. AB - Early stage Parkinson's disease may be better left untreated if it does not limit motor function. Once limitation of function is present levodopa-dopa decarboxylase inhibitor combinations are the most effective therapy, although amantadine may be satisfactory for a time in milder cases. The optimal independent roles of the ergot derivatives bromocriptine and pergolide, and the MAOb inhibitor selegiline, are not yet generally agreed although they are accepted as useful in supplementing the effects of levodopa. With prolonged levodopa use various late-stage treatment problems may appear. The pathogenesis of these is poorly understood and no completely satisfactory ways of managing them are available. PMID- 7487657 TI - Vaginitis. PMID- 7487658 TI - Practice tip. Dilating veins. PMID- 7487659 TI - 'Lumpy jaw' syndrome. PMID- 7487660 TI - Musculoskeletal medicine tip. Thoracic outlet. PMID- 7487661 TI - The unconscious patient. PMID- 7487662 TI - Shingles in a 63 year old woman. PMID- 7487663 TI - Two hours in the life of a government medical officer. PMID- 7487664 TI - A matter of balance. PMID- 7487665 TI - Back pain. PMID- 7487666 TI - The crown excision for facial lesions. PMID- 7487667 TI - Coding medical records. PMID- 7487668 TI - Knowledge of hepatitis C amongst general practitioners in the ACT. PMID- 7487669 TI - Unfamiliar terms need explaining. PMID- 7487670 TI - A device for controlling odorant stimulation and olfactory evoked responses in humans. AB - We developed a device for odorous stimuli control to record olfactory evoked responses from the human scalp. The characteristics of the apparatus are as follows. Translating the subject's respiration into electric signals with a sensor attached to the nose. The period and timing of odorous stimuli could be adjusted, so that stimuli could be synchronous with respiration. The interstimulus interval could be arbitrarily selected once every 1 to 9 respiration(s) so that adaptation could be prevented. We obtained evoked responses to odorous stimuli using this apparatus from the human scalp, whose positive peak latencies were approximately 350 and 700 msec. Such responses were not recorded if oxygen stimuli were used instead of odorous stimuli or with click sounds produced by the switching electromagnetic valve. Three types of odorant evoked scalp potentials were obtained in normal human subjects. The first type consisted mainly of two positive peaks with a peak latency of about 350 msec (P350) and about 700 msec (P700). The second type and the third type consisted of only one positive peak with a peak latency of about 350 msec and 700 msec, respectively. Such a P350 or P700 peak as that observed in the normal subjects was not detected in anosmic patients. PMID- 7487671 TI - Experimental observation of particulate flow along cranial nerves. AB - The purpose of this study was to investigate the communicative routes via the cranial nerves among the middle ear, the auditory tube, the inner ear, and the subarachnoid space under nearly physiological conditions. Various volumes of Indian ink were slowly injected into the facial nerve trunk of rabbits at the stylomastoid foramen, then serial sections of the heads were observed under the light microscope. This study yielded the following results: (i) Indian ink penetrated into the subarachnoid space through the internal auditory canal and/or the jugular foramen along the nerve sheaths of the VIIth, IXth, and/or Xth cranial nerves. (ii) Indian ink penetrated through the nerve sheaths of the cochlear, superior vestibular, and inferior vestibular nerves to the fundus of the internal auditory canal. (iii) Indian ink reached the scala tympani and the scala vestibuli via the cochlear aqueduct. (iv) Indian ink penetrated to the nerve sheaths of the tensor tympani and/or tensor palati nerves from the subarachnoid space of the trigeminal ganglion. In conclusion, it was shown that certain small particles are able to penetrate from the facial nerve sheath at the entrance of the stylomastoid foramen to the VIIIth, IXth, and Xth nerve trunks, the middle ear, the area around the auditory tube, the inner ear, and the subarachnoid space. PMID- 7487672 TI - Productivity of thyroglobulin and thyroid hormone on human thyroid cell in collagen gel culture. AB - Thyroid cells obtained from 21 normal human thyroid tissue samples and 17 tissue samples from diseased thyroid, including one of Graves disease, 4 with follicular adenoma, 11 with papillary carcinoma, and 1 with follicular carcinoma, were cultured in collagen gel, and ability to produce thyroglobulin (Tg), triiodothyronine (T3) and thyroxine (T4) was determined. Changes in morphological characteristics and production of Tg, T3, and T4 induced by addition of thyroid stimulating hormone (TSH) to medium in collagen gel culture were also determined. Twenty of all cases exhibited positive reaction for Tg. No relationship was found between rate of positivity for Tg and pathologic diagnosis. Three with carcinoma showed positive reaction for T3, and 4 with carcinoma showed positive reaction for T4. Only for normal thyroid cells did addition of TSH to medium induce increase the percentage of colonies producing Tg or T4 and morphological changes including an enlarged follicular lumen and increase in the height of columnar epithelium. These findings suggest that thyroid cells in collagen culture develop in an in vivo-like fashion. In conclusion, collagen has important effects on cellular differentiation when included in extracellular matrix. PMID- 7487673 TI - Cartinoid tumor of the middle ear and mastoid. AB - Primary cartinoid tumors in the middle ear and mastoid are rare. They are also very difficult to distinguish from adenomas and adenocarcinomas, using conventional histological stains. We present clinical, histological, immunohistochemical and ultrastructural findings of a cartinoid tumor in the middle ear and mastoid in a 40-year-old male. A soft tumor was revealed in the posterior mesotympanum and mastoid cavity, and a radical tympanomastoidectomy was performed. The tumor cells were stained by chromogranin A, and neurosecretory granules were confirmed with electron microscopy. We also review 20 previously reported cases in regard to their presentation, symptoms, signs, tumor extension, treatments, and histopathology. PMID- 7487674 TI - [Neurilemmoma of intra-temporal facial nerve:case report]. AB - A case of facial nerve neurilemmoma within the temporal bone is reported. The patient is a 47-year-old female complaining of 10 years' history of left facial palsy. CT and MRI examinations showed an enlarged mass of the tympanic cavity protruding into the petrous bone, mastoid cavity and external auditory meatus. An open biopsy of the tumor at the external auditory meatus revealed neurilemmoma. The tumor was totally removed by a retroauricular approach. Five months after surgery, both the cross nerve graft using the peroneal nerve and the temporal muscle transfer, method modified by Johnson were performed for the functional recovery of facial palsy. PMID- 7487676 TI - Effect of ibuprofen treatment during experimental acute otitis media. AB - In this study, the efficacy of concurrent treatment of experimental acute otitis media with ibuprofen and ampicillin was evaluated in chinchillas with respect to clearance of the effusion, presence of mucosal inflammation, and modulation of biochemical markers. Forty chinchillas were infected with non-typable Haemophilus influenzae and randomly assigned to treatment with either IM ampicillin (control) or ampicillin plus ibuprofen beginning on day 2 post inoculation. On days 5 and 10, animals from each group were killed, effusions recovered for biochemical assay, and the middle ears prepared for histological study. Differences in outcome measures favoring the control group were observed at the 5 day endpoint. However, at the 10 day endpoint, mucosal thickness was significantly less, the number of effusion free ears greater, and the concentrations of free fatty acids and thromboxane less in the animals treated with the combined therapy when compared to the control group. These results suggest that the addition of ibuprofen to ampicillin for the treatment of acute otitis media alters disease pathogenesis in this infectious model. PMID- 7487675 TI - Entomophthoromycosis caused by Conidiobolus coronatus: clinicomycological study of a case. AB - A case of midfacial swelling in a middle aged woman proved to be caused by a fungus Conidiobolus coronatus has been reported. It is a very rare condition and the knowledge of this fungus affecting the rhinofacial region in human started since 1965 only. These cases pose diagnostic problem. The treatment with the combination of ketoconazole and the saturated solution of potassium iodide was effective for this fungal infection. PMID- 7487677 TI - Early inflammatory changes of the Haemophilus influenzae-induced experimental otitis media. AB - Haemophilus influenzae is one of the most frequent pathogens of acute otitis media. To determine the middle ear response during the early stage of acute inflammation, a small amount of H. influenzae was inoculated into the bullae of guinea pigs through the tympanic membrane. The bullae were harvested at 6, 12, 24, 36, and 48 hours after H. influenzae inoculation and washed with phosphate buffered saline (PBS). The number of viable H. influenzae and inflammatory cells, the concentrations of myeloperoxidase (MPO) and lysozyme in the washing suspensions were measured, and compared with those in PBS-inoculated control ears. The number of viable H. influenzae increased very rapidly from 6 to 12 hours after inoculation and remained stationary up to 48 hours. The number of inflammatory cells and the MPO concentration were significantly higher in the H. influenzae-inoculated ears than in the control ears from 12 to 48 hours after inoculation. The lysozyme concentration was already significantly higher at 6 hours in the H. influenzae-inoculated ears; the lysozyme was released in the middle ear before the accumulation of inflammatory cells and degranulation of MPO from inflammatory cells. The results indicated that inflammatory reactions were present already at 6 hours after bacterial inoculation, and were rapidly accelerated during the subsequent hours. Consequently, acute middle ear inflammatory responses were seen immediately following inoculation of viable bacteria, and these responses originated in direct responses of middle ear mucosa, and oxidative and non-oxidative neutrophil metabolic products, which may cause tissue injury. PMID- 7487678 TI - The effect of the sympathetic nervous system upon susceptibility to acoustic trauma. AB - This study was designed to determine whether the sympathetic nervous system exerts a protective or enhancing effect in acoustic overstimulation. The compound action potential of the cochlea (CAP) was recorded in guinea pigs while the cervical sympathetic nervous system (SNS) was electrically stimulated or after it was surgically eliminated. The stimulation or the elimination of the cervical SNS has no effect on the threshold of CAP. The threshold shift in CAP after acoustic overstimulation (110, 115, or 130 dB SPL for 10 min) was measured in the cervical SNS stimulation group, in the cervical SNS elimination group, and in the control group. When the animal was under insufficient sedation, there was no difference among these three groups. However, the CAP threshold shift was significantly smaller in the cervical SNS stimulation group than in the other two groups when the animals were sufficiently sedated. The cervical SNS stimulation had some protective effect on the susceptibility to acoustic trauma when the systemic SNS activity was suppressed. PMID- 7487679 TI - Effect of hemorrhagic hypotension on blood flow to the basal and upper turns of the cochlea. AB - The effect of hemorrhagic hypotension on blood flow in the cochlea was studied in guinea pigs. Using two laser Doppler probes, simultaneous measurements were made from the basal and upper turns of the cochlea. Hypotension induced by removal of blood resulted in reduction of cochlear blood flow. Cochlear blood flow, however, did not reduce the same extent as the extent of hypotension and the reduction of blood flow was observed to be more remarkable in the upper turns than in the basal turn. PMID- 7487680 TI - Analysis of auditory brainstem response waveforms derived ipsilaterally and contralaterally to monaural stimulation. AB - The auditory brainstem responses (ABRs) obtained from 28 normal healthy adults were studied using the ipsilateral and contralateral recordings from the positions of vertex and each mastoid to monaural stimulation. Each wave of ABRs recorded by ipsilateral and contralateral derivations to stimulation site showed slightly significantly differences. Comparing the ipsilateral and contralateral data, the latencies of waves II and V showed a slightly small reduction in the ipsilateral recording, while these of waves III and IV showed a small increase. Next, the distributions of potentials and latencies of waves II to V were investigated from ABRs situating different electrodes in mid-coronal array of the scalp and non-cephalic reference electrode on the seventh cervical vertebra (CVII). The results of latencies showed the reverse relation to the data obtained from the ipsilateral and contralateral recordings using the reference electrodes on each mastoid. These facts suggest that the comparable differences of latencies in the bilaterally recorded ABRs are explained by the pseudo-phenomena of differential recordings, which the phase delayed or advanced potential propagated to each mastoid being reference electrode position. PMID- 7487681 TI - Toxicology: why are we swinging the lead? PMID- 7487682 TI - A QA perspective--report on ACE inhibitor angio-oedema. PMID- 7487683 TI - Risk factors and primary prevention of stroke. PMID- 7487684 TI - Flucloxacillin hepatitis--an Australian epidemic. PMID- 7487685 TI - The efficacy of chelation therapy and factors influencing mortality in lead intoxicated petrol sniffers. AB - BACKGROUND: The use of chelating agents to treat patients with petrol sniffing encephalopathy has been controversial, since alkyllead additives in petrol are not chelatable. A high mortality has also been reported in hospitalised petrol sniffers. AIMS: (i) Evaluate the efficacy of chelating agents in mobilising lead for excretion and lowering blood lead; (ii) Review factors contributing to mortality in hospitalised petrol sniffers. METHODS: All males chelated between 1992-1993 were studied (n = 20). Blood and urinary lead were measured daily before and during chelation then twice weekly until discharge. Parenteral calcium disodium edetate (EDTA) and dimercaprol (BAL) were administered together, every six hours for five days, seven patients subsequently received oral D penicillamine until discharge. Clinical details were reviewed for eight patients with petrol sniffing encephalopathy who died between 1990-1994. RESULTS: Urinary lead excretion substantially increased during parenteral chelation (median excretion = 113 microM/5 days, compared with pre-chelation excretion = 1.1 microM/day). Median blood lead concentration fell from 4.83 microM/L (pre chelation) to 1.91 microM/L (post-chelation). D-Penicillamine did not appear to increase urinary lead excretion appreciably. All eight deaths followed sepsis; five from complications of aspiration pneumonia. CONCLUSIONS: Airway maintenance and management are crucial for survival in these patients. In the short-term, parenteral chelation was effective in mobilising lead for excretion and reducing blood lead in encephalopathic petrol sniffers and was comparable to cases of inorganic lead intoxication. However, as in the treatment of inorganic lead intoxication, the long-term efficacy of chelation for petrol sniffers remains controversial. Prevention strategies against petrol sniffing at a community level are recommended. PMID- 7487686 TI - Trends in the home management of non-fatal acute myocardial infarction in Perth, Western Australia between 1984 and 1993. AB - BACKGROUND: Trends in the incidence of acute myocardial infarction (AMI) that are based on admissions to hospital would be misleading if the proportion of non fatal infarctions treated at home changed over time. AIM: To estimate trends in the management at home of cases of non-fatal AMI in Perth between 1984 and 1993 in order to determine if the number of such cases has remained consistently small and can be neglected in studies of the incidence of AMI. METHODS: In 1989, 251 general practitioners in Perth responded to a postal questionnaire asking about management of cases of AMI at home in the preceding 12 months. In 1993, 288 general practitioners and 174 physicians responded to the same questionnaire. RESULTS: We estimate that 1.5% of all patients less than 65 years of age diagnosed with non-fatal AMI were managed at home in 1989. The corresponding figure for 1993 was 4.1%. A previous study in Perth in 1984 found that 3.9% of cases of non-fatal AMI in patients less than 65 years of age were managed at home. CONCLUSION: The management at home of cases of non-fatal AMI in people of working age in Perth has remained at a negligible level from 1984 to 1993. Thus a register based on admissions to hospital for AMI will accurately reflect trends in AMI in people of working age. PMID- 7487687 TI - Outpatient liver biopsy using ultrasound guidance and the Biopty gun is safe and cost effective. AB - BACKGROUND: The need for liver biopsy is increasing with the increasing availability of treatments for liver disease, in particular interferon. For this reason the procedure must be efficient and economical, as well as safe. AIMS: Firstly to assess the safety of outpatient liver biopsy when performed with ultrasound guidance and the 'Biopty' gun. Secondly, to compare the cost of an outpatient and inpatient procedure. METHODS: Two hundred and fifty consecutive liver biopsies performed over a two year period at St Vincent's Hospital, Melbourne were reviewed retrospectively. Indications, histological findings and complications were recorded. RESULTS: Seventy per cent of the biopsies were performed as an outpatient procedure. There were no major complications. Four patients experienced prolonged pain post biopsy. The average cost for an outpatient biopsy was $351, the cost of an inpatient (overnight stay) was $690. CONCLUSION: Outpatient liver biopsy is safe and cost effective, with minor complications in 1.6%. The routine use of ultrasound guidance and the Biopty gun is likely to contribute to these favourable results. PMID- 7487688 TI - Peripheral blood lymphocyte subset distribution and function in patients with Alzheimer's disease and other dementias. AB - BACKGROUND: Until recently, new data on immune aspects of Alzheimer's disease (AD) have suggested that some facets of AD pathogenesis may be immune related. However, the effects of dementia itself on immune function have not been considered. AIM: To compare the distribution of peripheral blood lymphocyte subsets and their function in patients with AD and other dementias. METHODS: Peripheral blood lymphocyte numbers, T cell subset distribution, proliferative responses to mitogens and suppressor cell assay were studied in a well characterised group of patients with AD, and compared to patients with other forms of dementia. Age and sex matched elderly controls were screened to exclude dementia, and young controls were medical, paramedical and laboratory staff. Analysis of variance (ANOVA) and student's test were used for statistical analysis. RESULTS: The CD8+ lymphocyte population was reduced in AD and in other forms of dementia, when compared with non-demented elderly and young controls. Concanavalin A induced lymphocyte transformation was reduced in all dementia groups and in elderly compared with young controls. The changes in T cell numbers and function were not specific for Alzheimer's disease, but were found also in other forms of dementia. PMID- 7487689 TI - Serum 25-hydroxyvitamin D3 is related to physical activity and ethnicity but not obesity in a multicultural workforce. AB - BACKGROUND: Recent research suggests that body vitamin D levels are decreased in coronary heart disease and diabetes, but it is unclear which cardiovascular risk factors are related to vitamin D status. AIMS: To examine the relation between vitamin D status and major cardiovascular risk factors. METHODS: Serum 25 hydroxyvitamin D3, a marker of recent sun exposure and vitamin D status, was measured in 390 New Zealand residents (95 Pacific Islanders, 74 Maori and 221 others mostly of European descent), who were part of a larger cross-sectional survey of a workforce (n = 5677) aged 40-64 years. RESULTS: Serum 25 hydroxyvitamin D3 levels were significantly lower in Pacific Islanders (mean (SE) = 56 (3) nmol/L; p = 0.0001) and Maoris (68 (3) nmol/L; p = 0.036) compared with Europeans (75 (2) nmol/L) after adjusting for age, sex and time of year. Also adjusting for ethnic group, 25-hydroxyvitamin D3 was higher in people doing vigorous (aerobic) leisure physical activities (71 (2) nmol/L; p = 0.0066) and moderate (non-aerobic) activities (68 (3) nmol/L; p = 0.12) compared with those who were inactive (63 (2) nmol/L). However, 25-hydroxyvitamin D3 was unrelated to body mass index, serum lipids, blood pressure or cigarette smoking. CONCLUSIONS: People with increased skin pigmentation, such as Polynesians, and people who are inactive, have decreased body levels of vitamin D; this might partly explain their increased risk of cardiovascular disease. PMID- 7487690 TI - Adjuvant chemotherapy for non-metastatic osteosarcoma of the extremities in two New Zealand cancer centres. AB - BACKGROUND: Adjuvant chemotherapy significantly improves survival of patients with non-metastatic osteosarcoma but most of the data come from trials conducted in major international cancer centres. AIM: To review the efficacy and toxicity of an adjuvant chemotherapy regimen used in two regional cancer centres in New Zealand. METHODS: Retrospective review of patients treated for non-metastatic high-grade osteosarcoma of the extremities. The regimen (POMA) consists of high dose-methotrexate 8 g/m2 and vincristine 1.5 mg/m2 (maximum 2 mg) on days 1 and 8 followed by folinic acid then doxorubicin 50 mg/m2 and cisplatin 100 mg/m2 on day 15. This cycle was repeated every 35 days. Following amputation patients received six cycles while in selected patients two cycles were planned prior to limb salvage surgery followed by a further four cycles. Actuarial survival was calculated using the Kaplan-Meier method. RESULTS: Twenty patients were treated with POMA between 1986 and 1993. Amputation was performed in 16 patients and limb salvage surgery in four. Sixteen patients (80%) remain alive with no evidence of disease at a median follow-up of 40 months. Thirteen patients (65%) have been continuously disease-free. Actuarial survival at five years is 70%. Seven patients relapsed, six in lungs, of whom four underwent pulmonary metastasectomy; three of these remain free of disease 31, 35 and 40 months later. There was no local relapse. The toxicity of POMA is significant but tolerable. CONCLUSION: The results obtained at two regional cancer centres in New Zealand using POMA compare favourably to those achieved in clinical trials performed at major international cancer centres. PMID- 7487691 TI - Experience of once-daily aminoglycoside dosing using a target area under the concentration-time curve. AB - BACKGROUND: Many centres are changing to once-daily aminoglycoside administration. However, proposed methods for this practice often have theoretical and practical difficulties. We have developed a method in which a target area under the concentration-time curve (AUC) is used instead of traditional peak and trough serum concentrations. AIMS: To analyse our experience with the target AUC method in the first 100 courses of once-daily aminoglycoside administration in the Christchurch, New Zealand hospitals. METHODS: Following a starting dose of 5-7 mg/kg, administered by 30-minute infusion, the AUC was calculated using two serum aminoglycoside concentrations taken at one and six-14 hours after the start of the infusion. Dose adjustment was made to correct for any difference between the calculated AUC and a target AUC (72-101 mg.1(-1).h). The method was assessed for practicality and precision in 100 courses of treatment. The incidence of aminoglycoside toxicity was documented. RESULTS: The mean final dose of 6.68 mg/kg, and AUC of 92.8 mg.1(-1).h, were significantly different from the mean starting dose and AUC of 5.67 mg/kg and 74.0 mg.1(-1).h, respectively. The method appeared to be more precise than empirical dosing at achieving the target AUC even though the final recommended dose had more variability than the starting dose. Although the study was uncontrolled, observed nephrotoxicity (2%) and ototoxicity (up to 6.9%) were no greater than expected from the results of other studies. There were no deaths related to antibiotic failure. CONCLUSIONS: The AUC method was practical, and more appropriate for once daily dosing than the conventional method of aiming for target peak and trough concentrations. Dose adjustment can be made before the next dose. PMID- 7487692 TI - Early detection, screening and surveillance for colorectal cancer. PMID- 7487693 TI - Change in zidovudine prescribing practices in Australia, 1991-1993. PMID- 7487694 TI - Failure to recognise the association of life-threatening angio-oedema and angiotensin-converting enzyme inhibitor therapy. PMID- 7487695 TI - Epidemiology of Jakob-Creutzfeldt disease in Australia 1970-80. PMID- 7487696 TI - Assessment of idiopathic dilated cardiomyopathy using positron emission tomography (PET). PMID- 7487697 TI - Public policy or personal gain--the politics of medical representation. PMID- 7487698 TI - A brave new world of cancer screening. PMID- 7487699 TI - Cancer screening and employment. PMID- 7487700 TI - Phaeochromocytoma associated with myasthenia gravis precipitated by propranolol treatment. PMID- 7487702 TI - Hereditary fructose intolerance in an adult. PMID- 7487701 TI - Warfarin interactions with Chinese traditional medicines: danshen and methyl salicylate medicated oil. PMID- 7487703 TI - Delayed neuroleptic malignant syndrome following cessation of prolonged therapy with metoclopramide. PMID- 7487704 TI - Methylene blue fails to improve hypoxaemia post orthotopic liver transplantation (OLT) PMID- 7487705 TI - Electrical injury: cervical myelopathy with late onset progressive motor neuron disease. PMID- 7487706 TI - Recurrent aseptic meningitis. PMID- 7487707 TI - Induction (neo-adjuvant) chemotherapy: systemic and arterial delivery techniques and their clinical applications. AB - Induction chemotherapy is a well tried method of improving the cure prospects of locally advanced or aggressive primary cancers. It is most simply administered by systemic (intravenous) delivery but for some cases a greater chemotherapy impact is both desirable and achievable with the use of a more concentrated regional delivery technique. Any advantage of regional chemotherapy will depend upon the type of tumour to be treated, its site and blood supply, the most effective doses, concentrations and exposure periods of the preferred agents to be used and the relative risks of systemic and regional toxicity and the techniques of delivery. In general, systemic chemotherapy is most appropriate in treating tumours without a single artery of supply; when certain agents which are inactive until modified in body tissues (such as cyclophosphamide or DTIC) are to be used; when satisfactory responses can be achieved more easily by systemic delivery; when technical skills and facilities for regional delivery are not available; or when the patient's general health, co-operation or long-term prognosis precludes the additional complexity of regional delivery. Intra-arterial infusion may have advantages in treating head and neck, gastric, liver, some locally advanced breast, limb, pelvic and possibly pancreatic malignancies. More complex techniques to achieve short-term highly concentrated chemotherapy include closed circuit perfusion; chemofiltration infusion and 'stop flow' perfusion; and regional limb infusion. These should remain the subject of ongoing studies in highly specialized units which treat tumours such as melanoma or pancreatic cancer which respond poorly to lower chemotherapy concentrations. PMID- 7487708 TI - Multiple endocrine neoplasia type I: clinical syndrome to molecular genetics. PMID- 7487710 TI - Papillary thyroid carcinoma arising in thyroglossal duct cysts: incidence and management. AB - The incidence and pathological features of papillary thyroid carcinoma arising in the thyroglossal duct cysts were reviewed and compared with papillary thyroid carcinoma arising elsewhere in the thyroid gland. In the 30 year period 1964 to 1993 there were 90 thyroglossal duct nodules or cysts treated surgically at the Endocrine Surgical Unit, Royal North Shore Hospital, Sydney, Australia. There were four cases of papillary thyroid carcinoma in this group (4.4%). In the same period 2814 cases presented with clinical single thyroid nodules which were treated surgically. There were 182 cancers in this group of which 121 were papillary thyroid carcinomas (4.3% of total cases). This is identical to the incidence seen in the thyroglossal duct. We conclude that the incidence of papillary thyroid carcinoma arising in the thyroglossal duct is no different to that arising elsewhere in the gland. The difference in number of carcinomas related only to the volume of follicular thyroid tissue present in the gland proper. That being the case, there is no reason to treat these cancers differently from papillary thyroid carcinoma elsewhere in the gland. PMID- 7487709 TI - Inactivation of retinoblastoma gene in malignant parathyroid growths: a candidate genetic trigger? AB - The molecular basis for parathyroid carcinoma remains undetermined. Parathyroid carcinoma potentially remains curable by early en bloc resection. This requires a reliable diagnostic tool as histological features alone are insufficient to distinguish parathyroid carcinoma from its benign counterpart, parathyroid adenoma. A variety of human cancers arise from the inactivation of the retinoblastoma (RB) gene, a tumour-suppressor gene on chromosome 13q14. We investigated the role of this gene in parathyroid growths by using a mouse monoclonal antibody to detect RB gene expression immunohistochemically. Two of the three parathyroid carcinomas in this study showed evidence of RB gene inactivation compared with one of 11 benign parathyroid entities. Three normal parathyroid glands stained showed strong evidence of RB gene expression in all three glands. The relationship between RB gene inactivation and parathyroid malignancy, however, was not statistically significant. PMID- 7487711 TI - Gastric carcinoma with synchronous liver metastases: palliative gastrectomy or not? AB - During a 6 year period, 38 patients with gastric cancer presented with synchronous liver metastases and 25 underwent gastric resection. Forty per cent of the operated patients presented with complications, whereas most of the non operated patients presented with pain or an abdominal mass. Eight patients (32%) developed postoperative morbidity, five of whom died (20%). The median duration of hospital stay for those surviving surgery was 33 days. In the non-operated group 61% died while in hospital and the median duration of hospital stay was 28 days. The respective median survival time and duration of home stay were 13 and 9 weeks for the operated patients and 6 and 3 weeks for the non-operated patients. The difference of the duration of home stay between the two groups of patients was statistically insignificant. There was also no significant relief of pain after surgery. Univariate analyses of the influence on survival time of operation among 13 other factors showed that only bilirubin was significant. We conclude that gastrectomy neither prolongs life nor improves the quality of survival in patients with gastric cancer and discontiguous liver metastases. However, gastrectomy may be beneficial in selected patients presenting with potentially lethal complications such as bleeding and obstruction. PMID- 7487712 TI - Carcinoma of the gall-bladder: an experience and review of the literature. AB - This paper summarizes the treatment results of 28 cases of gall-bladder carcinoma at Toowoomba Base Hospital, Toowoomba, Queensland, Australia. The literature is reviewed in order to determine whether more extensive resection is improving the outcome of a disease generally understood to have a deplorable prognosis. The complete records of 28 patients with this malignancy were obtained from the period 1978 to 1994 and analysed retrospectively. Twenty-three patients had cholecystectomy, but only one underwent formal lymph node dissection. Overall, the 5 year survival rate was 13%, and long-term survivors all had stage I or II disease. Those with stage III or IV disease had a median survival of 46 days. Recent literature confirms that simple cholecystectomy is not always curative for early lesions discovered incidentally and also that some patients with advanced disease can achieve long-term survival if treated with radical resection. Wedge resection of the liver and nodal clearance of the hepatoduodenal ligament at least has been recommended in patients with T2 lesions (subserosal/perimuscular), and in selected patients with stage III disease. PMID- 7487713 TI - Resection and anastomosis of obstructed left colonic cancer: primary or staged? AB - A retrospective analysis of the treatment of 70 patients with obstructed left colonic cancer was undertaken in order to assess whether staged or primary resection was more appropriate. Thirty-four patients had initial colostomy and staged resection (group 1) while 36 patients were treated by primary resection and immediate anastomosis following intra-operative bowel washout (group 2). There were seven deaths (10%), five in group 1 and two in group 2. The wound infection rate and average hospital stay were 44% and 36 days in group 1 and 19.4% and 16.5 days in group 2, respectively. Twenty per cent of patients in group 1 did not complete their staged procedures and had to live with their colostomies. The smoother postoperative recovery and shorter hospital stay was particularly significant in group 2 patients. As primary resection and anastomosis can now be performed with relative safety and reduced morbidity, we conclude that staged procedures can no longer be accepted as standard treatment for left colonic obstruction. PMID- 7487714 TI - Early experience with postoperative adjuvant chemoradiation for rectal carcinoma: focus on morbidity. AB - The morbidity of postoperative adjuvant chemoradiation for primary extraperitoneal rectal carcinoma is documented in this ongoing study. Patients who presented electively for resection with ACPS Dukes' B and C extraperitoneal rectal carcinoma during the period January 1990 to June 1993 were studied. Twenty patients received postoperative adjuvant chemoradiation. At a mean follow up of 32.6 months (s.d. 7.1), three of the 20 patients who received combined adjuvant chemoradiation reported no side effects. In 10 patients (50%) complications were classified as minor. In the remaining seven patients (35%) major complications of therapy occurred. There were no deaths. These early results highlight the morbidity of postoperative adjuvant chemoradiation that has been presumed but not documented. PMID- 7487715 TI - Video-assisted thoracoscopic wedge resections of pulmonary metastatic osteosarcoma: should it be performed? AB - We studied the use of video-assisted thoracoscopic (VAT) surgery in the management of metastatic osteosarcoma. From September 1993 to March 1994, we performed a total of 11 VAT wedge resections of pulmonary metastatic osteosarcoma in seven patients (six males, one female, age 12 to 46 years). Three patients had bilateral procedures performed either under the same anaesthesia or in stages. One patient had two operations on the same side. The average number of nodules excised was three. Two patients subsequently required formal lobectomies when the metastatic tumours were either too big or too close to the hilum for safe wedge resections. There was one death on postoperative day 3 due to dysrhythmia. One patient died 5 months later from a progression of his underlying disease. Two patients remained disease free up to 8 and 12 months, respectively, from their first operations. The average postoperative chest drain duration was 1.4 +/- 0.7 days and hospital stay 2.3 +/- 1.1 days. The procedure was well tolerated and postoperative morbidity was minimal. We conclude that although VAT wedge resection of pulmonary metastatic osteosarcoma is feasible technically and is associated with a short hospital stay and minimal morbidity, this approach cannot be recommended when complete resection of all metastases is the goal as the technique relies heavily on computed tomographic scans to detect nodules. Recurrence of metastasis from 4 to 6 months in three of seven patients argues against VAT surgery being an adequate procedure. The high cost of the staplers, in addition, is a secondary consideration. PMID- 7487716 TI - Immunohistochemical detection of p53 and the recurrence of rectal adenocarcinoma. AB - The relationship between immunohistochemical detection of the p53 tumour suppressor gene and recurrence of disease in 66 cases of rectal adenocarcinoma treated at the Royal Hobart Hospital, Hobart, Tasmania, between 1984 and 1992 was investigated. Fifty-five per cent (n = 36) of patients demonstrated p53 nuclear immunoreactivity consistent with the presence of the stabilized protein derivative of a mutation at the p53 gene locus. No correlation between p53 immunoreactivity and tumour site, differentiation and stage was noted. An overall recurrence rate of 64% (n = 42) was noted. Multivariate analysis confirmed the absence of p53 immunoreactivity and anatomical site of tumour below 7 cm from the anal verge to be significant predictors of tumour recurrence (P < 0.001). A review of the current literature on p53 suggests caution in using the immunohistochemical detection of p53 to predict the clinical outcome in cases of rectal adenocarcinoma because of discordant results between studies. PMID- 7487717 TI - Laparoscopic splenectomy. AB - The innovations in laparoscopic technology have expanded the variety of general surgical procedures amenable to laparoscopic approach. An initial experience with eight cases of laparoscopic splenectomy is presented. The indications for splenectomy were immune thrombocytopenic purpura (ITP, n = 6), hereditary spherocytosis (n = 1) and enlarged spleen with filling defects (n = 1). There were six females and two males, aged 27 to 46 years. Seven patients had the spleen removed laparoscopically and one had laparoscopically assisted splenectomy. The operations took from 2 h 15 min to 3 h 30 min (mean 2 h 45 min). The spleens removed varied from 70 to 563 g (mean = 250). Blood loss ranged from insignificant to 1400 mL. Significant bleeding was encountered in three patients with moderate splenomegaly (240, 350, 563 g). Two patients received autologous blood transfusion. The average narcotic required was three doses. The patients were discharged after a mean postoperative stay of 3 days (range 2-4). One patient developed a below knee deep venous thrombosis. Laparoscopic splenectomy is possible and promises to provide the advantages associated with other laparoscopic procedures. Patients with an enlarged spleen can present a technical challenge, and there is a potential for significant blood loss. The current technology and laparoscopic expertise means that this procedure should probably be limited to patients whose spleens are not palpable clinically. PMID- 7487718 TI - Laparoscopic splenectomy using the lateral position: an improved technique. AB - An alternative approach to laparoscopic splenectomy is described. This technique uses a lateral position and modified port placement. Initial experience with three patients has demonstrated advantages over previously described approaches, by providing a more direct view of the splenic hilum and by eliminating the need for retraction of adjacent organs. PMID- 7487719 TI - Frederic Wood Jones: coral and atolls. PMID- 7487720 TI - Meigs' syndrome in association with a locally invasive adenocarcinoma of the fallopian tube. AB - A patient with recurrent pleural effusions posed a diagnostic challenge. The pleural aspirate CA-125 assay was markedly elevated which indicated a primary ovarian pathologic process. However, in this case a rare fallopian tube adenocarcinoma was the underlying problem. Meigs' syndrome can resemble a metastatic pelvic carcinoma in its presentation, but the prognosis is much more favourable when the syndrome is associated with a benign ovarian tumour. PMID- 7487723 TI - Classification and grading of chronic venous disease in the lower limbs: a consensus statement. American Venous Forum. PMID- 7487721 TI - Phlegmonous enterocolitis: a rare form of inflammatory bowel disease. AB - Phlegmonous enterocolitis is a rarely encountered and almost universally fatal bacterial infection seen most often in the alcoholic population. Despite being first described in 1842, fewer than 60 cases have been reported. The role of surgical management in this condition is unknown. We present a case of phlegmonous enterocolitis with a successful outcome after total colectomy. PMID- 7487722 TI - Laparoscopic pelvic lymphadenectomy for malignant melanoma. AB - Therapeutic laparoscopic techniques are well accepted in many areas of general surgery. The use of laparoscopy for the staging of malignancy, in a diagnostic role, is not used widely. Urologists use pelvic lymph node dissection (LPLND) to define those patients who are suitable for curative local therapy in carcinoma of the prostate. We present a case of a patient with metastatic malignant melanoma in whom LPLND was employed as a minimally invasive way of staging and treating her disease. PMID- 7487724 TI - Modern day treatment of acute deep venous thrombosis. AB - Acute deep venous thrombosis is a spectrum of disease presentations. Proper treatment mandates precision in diagnosis and knowledge and availability of therapeutic alternatives. Natural history studies and therapeutic trials have indicated that thrombus resolution (elimination) is associated with improved long term functional outcome. The preferred approach to most patients with proximal venous thrombosis is to eliminate thrombus either pharmacologically or mechanically, to provide unobstructed venous drainage to the affected extremity and to offer subsequent treatment to ensure that recurrent thrombosis does not occur. If these principles can be followed without placing the patient at an unacceptable risk of a complication, substantial strides can be made in the treatment of patients with acute DVT. PMID- 7487725 TI - Adenosine thallium-201 scans in patients undergoing elective non-cardiac surgery. AB - Dipyridamole Thallium-201 (201Tl) scintigraphy has been used widely for assessment of patients prior to vascular surgery. Recently, Adenosine has been reported to be a safe and useful alternative to Dipyridamole. The purposes of this study were to evaluate the safety and feasibility of the use of Adenosine, to evaluate the correlation of Adenosine 201Tl scans with coronary angiograms (when available) and to evaluate the effect of scan results on clinician management style. Fifty adults with abdominal aortic aneurysm or other vascular disease underwent an intravenous infusion of Adenosine in conjunction with initial and delayed planar 201Tl scans. Images were interpreted qualitatively and quantitatively by the consensus of two or more experienced observers with patients showing transient left ventricular dilatation or redistribution in one or more myocardial segments reported as being at high risk of peri-operative cardiac events. Of the 50 subjects studied, 49 tolerated the maximum infusion dose with 60% experiencing minor transient symptoms. Low (n = 30) and high risk (n = 20) patients were defined according to Adenosine 201Tl scans. Age, gender and clinical characteristics were similar in both groups. Thirteen (65%) high risk subjects had coronary angiography compared with only three (10%) low risk patients. Patients with high-risk 201Tl scans were also more likely to proceed to coronary revascularization prior to non-cardiac surgery [5/20 (25%) vs 1/30 (3%)]. The positive predictive value of high risk 201Tl scans for coronary artery disease was 85%. Thus, Adenosine is considered a useful and safe alternative to Dipyridamole.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7487726 TI - Duplex ultrasound assessment of the venous status of the swollen leg. AB - Between January 1991 and December 1993, duplex ultrasound characterization of venous disease in leg swelling was studied in 214 patients (261 limbs; 167 unilateral and 47 bilateral). All patients were examined with a duplex scanner, the superficial and deep venous systems were evaluated for the presence of thrombus and valvular incompetence. Of the 261 limbs, 29 (11.1%) had deep venous thrombosis, 14 (5.4%) had superficial venous thrombosis, 66 (25.3%) had deep venous incompetence (31/66 limbs also had superficial venous incompetence), 65 (24.9%) had incompetence in the superficial veins only, and five (1.9%) had deep venous obstruction resulting from a popliteal cyst or a popliteal vein ligation. Eighty-two limbs (31.4%) had no evidence of venous obstruction or incompetence at the areas evaluated. This study showed that venous obstruction and valvular incompetence had occurred in two-thirds of swollen legs examined. Some of the venous obstructions resulted from surgically treatable diseases such as a popliteal cyst, and some of the venous disorders involved the superficial venous system only. Complete venous evaluation with duplex imaging can be very helpful in the determination of the underlying cause of the swelling. PMID- 7487727 TI - Application of Doppler/ultrasound examination for varicose veins in a provincial hospital. AB - There are a number of different primary problems in patients presenting with varicose veins. Treatment needs to be appropriate to the different causes. Doppler/ultrasound examination of varicose veins can very accurately define almost all abnormalities that cause varicose veins, but this can be time consuming. Over 3 years' experience of the clinical application of colour Doppler/ultrasound investigation in the assessment of patients with varicose veins in a small general hospital is reported. The selection of patients and how improved understanding of the various presenting problems of patients examined may affect treatment are described. All surgeons with access to this technology are encouraged to include it in the investigation of their patients. PMID- 7487728 TI - Heparin thromboprophylaxis via indwelling subcutaneous teflon cannula. AB - The aim of this study was to evaluate the administration of postoperative heparin thromboprophylaxis via an indwelling subcutaneous cannula. Fifty consecutive patients undergoing laparotomy, laparoscopy or varicose vein surgery were studied. Each patient on admission had a 24 gauge subcutaneous cannula inserted and a conventional subcutaneous heparin injection preoperatively. The first 20 patients received an alternating postoperative regimen of morning heparin via cannula and evening heparin via conventional injection into the abdominal wall. The remaining 30 patients received postoperative heparin twice daily via the cannula. Patients and nursing staff were interviewed and their preference for either method was recorded. There were no significant complications with the cannula system. Although it was more costly and nursing staff found it more labour intensive than conventional subcutaneous injections, the majority of patients preferred to receive heparin via the cannula. We conclude that this is an excellent method of administering prophylactic subcutaneous heparin, especially for patients with a strong aversion to injections. PMID- 7487729 TI - The risk factors in central venous catheter-related thrombosis. AB - Catheter-related venous thrombosis is one of the most important complications of central venous catheters. The aim of this prospective study was to assess the risk factors that may be important in the development of catheter-related thrombosis. Multiple lumen (n = 20) and single lumen (n = 24) polyurethane catheters were inserted to the subclavian vein by the infraclavicular approach in 44 consecutive patients. All variables that may be significant for the development of thrombosis related to the patient, the catheter, the insertion technique and the solution administered through the line were recorded. Eighteen of 44 patients (40%) were found to have venous thrombosis. Two factors were found to correlate significantly with the development of central venous catheter related thrombosis, namely the number of vein punctures (one vs two punctures, P < 0.01) and the composition of solution (fluid replacement vs total parenteral nutrition, P = 0.01). PMID- 7487730 TI - Pregnancies complicated by acute appendicitis. AB - Appendicitis is one of the most common acute abdominal conditions that requires operation during pregnancy. The maternal and fetal complications occurring in such situations are well known. However, the diagnostic and therapeutic challenges of acute appendicitis complicating pregnancy persist in current surgical practice. Thirty-eight obstetric cases who had an appendicetomy performed during pregnancy between 1986 and 1993, were reviewed. In 31 cases, acute appendicitis was proven at laparatomy, and by subsequent histopathology. In the confirmed cases, seven occurred in the first trimester, 12 in the second trimester, eight in the third trimester and four in the immediate postpartum period. Two miscarriages occurred in the first trimester group, and two late abortions and two preterm (26 and 28 weeks) deliveries occurred in the second trimester group. In the third trimester/postpartum group, all babies survived, but four patients had Caesarean sections with the appendicetomy and the incidence of perforated/gangrenous appendices was highest. Perinatal outcome was worst, with the highest fetal wastage rate if the acute appendicitis occurred in the second trimester, although maternal morbidity appeared to be the same across different gestational periods. PMID- 7487731 TI - Gastric devascularization and splenectomy for bleeding gastric varices. AB - Gastric varices may be associated with oesophageal varices in patients with portal hypertension. Control of haemorrhage from gastric varices is often difficult. Between June 1988 and February 1990, six patients underwent gastric devascularization in an effort to control life-threatening haemorrhage from bleeding gastric varices after non-operative measures failed. In all six patients the bleeding was controlled. There was one peri-operative death due to hepatic failure. Four patients experienced postoperative respiratory or urinary tract infections that responded to appropriate antibiotic and respiratory support measures. After a mean follow-up period of 44 months, eradication of gastric varices was achieved in all five survivors. Gastric devascularization is an effective surgical measure for good palliation and control of life-threatening haemorrhage from bleeding gastric varices. PMID- 7487732 TI - Complications of an abdominal stoma: an analysis of 322 stomas. AB - This paper reports a series of 316 patients with 322 stomas, of which 156 were end-sigmoid colostomies (48.5%) and 123 urological ileal conduits (38.2%). An overall complication rate of 66.8% was detected, with parastomal herniation rate, stenotic rate, and prolapse rate of 31.1, 10.2 and 6.8%, respectively. Complications were detected much later in a urological stoma than in a colostomy. Patients bearing an abdominal stoma should be followed up stringently. The creation of an abdominal stoma should not be regarded as a minor surgical procedure. Certain stomas, such as loop transverse colostomy, should be avoided whenever possible. PMID- 7487733 TI - Issues in biomedical statistics: comparing means by computer-intensive tests. AB - In this review there are described two alternatives to classical tests for distinguishing means. These are called computer-intensive because they can only be performed on fast computers. Permutation procedures have the virtue in that they are easy to understand, they can be employed to analyse small sets of experimental data, and under the randomization model of inference (though not the population model) they require no assumptions except that the experimental groups have been constructed by randomization. Bootstrap procedures are designed for use under the population model of inference (though not the randomization model) and are best suited to larger sets of experimental data. Non-parametric bootstrapping requires populations to be sampled randomly, but it depends on no prior assumptions about the distributions of those populations. It is argued that if randomization rather than random sampling has been done, permutation tests are superior to the classical t and F tests for detecting differences between means and therefore should replace them. If random sampling has been done, non parametric bootstrap techniques may prove to be superior to classical tests for constructing population confidence intervals or testing hypotheses. However, their accuracy, especially for hypothesis-testing and when samples are small, has yet to be firmly established and there is a dearth of commercial software with which they can be executed on personal computers. PMID- 7487734 TI - Home-made videoscopic trainer for operative laparoscopic surgery. AB - The author has used this home made videoscopic trainer for more than 12 months and found it very helpful and practical for refining laparoscopic skills. It is strongly recommended to all surgeons and trainees embarking on laparoscopic work. PMID- 7487735 TI - Changes in practice and knowledge in orthopaedic surgery. PMID- 7487736 TI - Cutaneous vasculitis and cutaneous vasculopathies. AB - Cutaneous involvement may occur with virtually all syndromes of vasculitis. This can occur primarily as a dermatologic disorder or as a manifestation of a potentially life threatening systemic vasculitis. In this review article, classification, clinical manifestations, pathogenesis and therapy of cutaneous vasculitis will be discussed. Disorders which are primarily vascular in origin but lack a well defined inflammatory phase, referred to as 'vasculopathies' will also be discussed. PMID- 7487737 TI - Mechanisms of drug eruptions: Part I. AB - The pathogenetic mechanisms underlying common, and less common but severe, adverse cutaneous drug reactions are reviewed. Pharmacogenetic variability may account for a susceptibility to serious drug reactions to sulphonamides and anticonvulsants, as well as to lupus erythematosus (LE)-like syndrome. Exanthematous drug reactions may have an immunological basis. Cell mediated cutaneous drug reactions, including lichenoid reactions, LE-like syndrome, fixed drug eruption, erythema multiforme, Stevens-Johnson syndrome and toxic epidermal necrolysis, will inevitably involve elements of the skin immune system. Graft versus-host disease provides a useful model for aspects of these drug-induced disorders. Urticaria, angioedema, anaphylaxis and anaphylactoid reactions may involve Type I immunoglobulin (Ig)-mediated or Type III hypersensitivity, or may be caused by pharmacological, non-allergic means. Drug-induced vasculitis, serum sickness and the Arthus phenomenon are manifestations of the immune complex disease. Drug-induced pemphigus may involve immune dysregulation, but several thiol-containing drugs are able to cause antibody-independent acantholysis directly. PMID- 7487738 TI - Conducting a factory or plant visit. AB - Plant visits are an invaluable tool in the management of work-related skin disorders, as they are the means by which a dermatologist gains firsthand information about the circumstances under which a work-related skin disorder has arisen in a plant or factory. In order to manage the workplace hazards which are responsible for skin disease, these must be identified and evaluated, and recommendations made about their control. While it may be possible to do this vicariously by questioning, examining, and investigating affected workers in the office, site visits to the plant or factory are sometimes necessary for the optimal assessment and management of such cases, as they often provide valuable information and insights not forthcoming from the history, examination, and investigations. This article explores the rationale behind plant visits, and discusses the pre-visit assessment, pre-visit preparation, the plant visit itself, and post-visit responsibilities. Practical advice is given for conducting each of these stages. PMID- 7487740 TI - Quantification of hair follicle parameters using computer image analysis: a comparison of androgenetic alopecia with normal scalp biopsies. AB - Computer image analysis enables large numbers of hairs to be measured in an automated fashion. In this study, we examined horizontal scalp biopsies from 10 patients with a histological diagnosis of androgenetic alopecia and 10 normal control subjects. The density of hair follicles and the ratio of terminal to vellus hairs were determined. Hair shaft, hair canal and hair follicle diameter, inner root sheath width and outer root sheath area were measured using the Chromatic Colour Image Analysis program. This study showed a statistically significant progressive decrease in size of hair canal diameters from normal terminal hairs (85.93 +/- 10.07 microns) through to androgenetic alopecia terminal (68.83 +/- 13.60 microns) and vellus hairs (28.67 +/- 5.60 microns). This pattern is also seen with hair follicle diameters; normal terminal (268.41 +/- 24.88 microns), androgenetic alopecia terminal (236.34 +/- 17.23 microns), and vellus hairs (130.88 +/- 19.96 microns). Outer root sheath areas, hair shaft diameters and ratio of terminal to vellus hairs were significantly larger in normal (18,500 +/- 4222 microns 2; 82.71 +/- 13.79 microns; 36:1; respectively) compared with androgenetic alopecia scalp biopsies (8403 +/- 3322 microns 2; 61.11 +/- 14.42 microns; 3:1; respectively), whereas inner root sheath width and density did not vary significantly. Computer image analysis can be adapted for use in clinical trials where large numbers and objectivity are critical in determining the efficacy of hair growth promoters. PMID- 7487739 TI - Cryosurgery of malignant and premalignant diseases of the skin: a simple approach. AB - Cryosurgical treatment of skin cancer and premalignant conditions of the skin has been in widespread use for 20 years. Data accumulated over this period suggest that if attention is paid to the treatment technique and to lesion selection, then cure rates equivalent to radiotherapy, simple surgical excision, and curettage and cautery can be achieved reliably. Moh's micrographic surgery offers a higher cure rate for skin cancer, but is not suitable for the vast majority of lesions seen in clinical practice. The decision to use cryosurgery to treat any particular lesion will therefore be influenced by a number of other considerations. Cryosurgery competes well on morbidity and cosmetic outcome and is the quickest, easiest, cheapest and most readily available of the treatment options. As such it has earned its place among the recognized treatment modalities for skin cancers as well as premalignant conditions of the skin. Cryosurgery is commonly delivered empirically without record of the dose delivered and without audit of the outcome. The aim of this review is to describe in detail one standard technique of therapy that is easily reproduced and has been audited; the timed spot freeze technique. This technique can be used, even by those inexperienced in cryosurgery, to achieve predictable success rates. Many other techniques do exist, but either have not been audited or are unnecessarily cumbersome. PMID- 7487741 TI - Pale grain eumycetomas in Madras. AB - Biopsy specimens from 211 cases of mycetoma were examined histologically. Pale grain eumycetoma was found in seven cases. Four of these were studied mycologically, Acremonium kiliense was isolated from two and Acremonium falciforme and Pseudallescheria boydii from one case each. The geographic distribution of these organisms, and their incidence and prevalence are discussed. PMID- 7487742 TI - Ehlers-Danlos syndrome type II: importance of recognition. AB - We report a case of Ehlers-Danlos syndrome Type II presenting with subtle clinical features and discuss the importance of early recognition of this disorder. A 16 year old Caucasian schoolgirl presented with soft tissue swellings on her fingers. There was a history of poor wound healing and prematurity and a family history of miscarriages. Clinical examination revealed hypermobile joints, hyperelastic skin and a 'cigarette paper' scar at the site of a previous minor surgical excision. Management consisted of genetic counselling and advice concerning career choice, recreational activities, as well as future pregnancy and surgery. This subtype of Ehlers-Danlos syndrome is probably fairly common in the community. While it may present with subtle clinical features, recognition is important so that counselling can be offered at a young age in order to avoid the major consequences that may arise during pregnancy and surgery. PMID- 7487743 TI - Transient zinc deficiency in a breast-fed premature infant. AB - Symptomatic zinc deficiency developed in a breast-fed premature male infant of 31 weeks gestation. At 13 weeks of age he presented with diarrhoea, irritability and an eruption identical to acrodermatitis enteropathica. Breast milk zinc concentrations were low. His course was complicated by milk protein intolerance. After 7 weeks, zinc supplementation was ceased without recurrence of disease. PMID- 7487744 TI - Lymphocutaneous sporotrichosis. AB - Lymphocutaneous sporotrichosis presented in a 10 year old child 1-2 weeks after an abrasion. A series of nodules, two of which ulcerated, appeared along the arm with tender unilateral axillary lymphadenopathy in the absence of systemic symptoms. Biopsy showed a granulomatous infiltrate but failed to reveal the organism; however, culture was positive for Sporothrix schenckii. The primary lesion healed with a scar after 3 months of systemic therapy with potassium iodide. PMID- 7487745 TI - Polydactylous Bowen's disease of the nail bed. AB - A case is presented with tender swelling and nail bed hyperplasia in 2 fingers. Histopathology revealed Bowen's disease in both fingers and after amputation, invasive squamous cell carcinoma in one finger. PMID- 7487747 TI - Chromate allergy from 'antifreeze-antiboil'. AB - A diesel mechanic developed hand dermatitis after more than 3 years in the trade. Patch testing showed positive reactions to chromate and cobalt. Potassium dichromate is added to antifreeze-antiboil as a corrosion inhibitor. PMID- 7487746 TI - Lichen sclerosus in childhood. AB - Lichen sclerosus in childhood involves predominantly the genital area, often with purpura and erosions, which raises concerns of sexual abuse. Potent topical corticosteroids give a good clinical response over several months, although maintenance with weaker formulations is usually required. The long-term prognosis remains uncertain. PMID- 7487748 TI - The recovery suture. AB - Continuous running sutures have several advantages in wound closure but can be difficult to remove. A loop tied around an exposed section of the running suture or an adjacent interrupted suture allows visualization and ease of removal. PMID- 7487749 TI - Discoid erythematosus in Australian aborigines. PMID- 7487750 TI - Breast cancer following treatment for Hodgkin's disease. PMID- 7487752 TI - The changing face of radiation oncology in Australia 1950-1995: a personal view. AB - In 1950, orthovoltage X-rays were used to treat a wide spectrum of malignant disease but results were impaired by poor depth dose and significant reactions; radium and radon were used commonly and superficial X-ray therapy was used for a large number of skin cancers as well as many benign skin conditions. Since megavoltage X-ray therapy was introduced to Australia and with earlier diagnosis, high standards for qualification as a radiation oncologist set by the Royal Australasian College of Radiologists (RACR), improved imaging and tumour localization, improved beam characteristics, computerization and dosimetry, patients are now better treated with improved local control and less morbidity. Public facilities have been chronically underfunded by governments resulting in waiting lists for treatment, and free standing private practices are important in sharing the ever increasing workload consequent to the increasing and ageing population. The use of brachytherapy has fallen but is enjoying a resurgence of interest. Chemotherapy has emerged. Radiobiology has had some influence on radiation treatment. Health education has improved and the needs and expectations of patients better appreciated and helped. Cancer societies have made an important contribution. Statistical evaluation of treatment is better understood. Undergraduate training and academic positions remain insufficient. The formation of the Faculty of Radiation Oncology of the RACR has given fresh impetus to the specialty. PMID- 7487751 TI - The history of neuroradiology: an Australian perspective. PMID- 7487753 TI - Swollen ischiopubic synchondrosis: a dilemma for the radiologist. AB - Differences in size and shape of ischiopubic synchondrosis in childhood may present problems in diagnosis and differential diagnosis. Whereas asymptomatic swollen ischiopubic synchondrosis represents a normal ossification process, painful swelling is a symptom of underlying pathology. Five children are described with symptomatic ischiopubic synchondrosis swelling, four representing stress reaction and one with osteomyelitis. Radiologists should be careful when reporting on swollen ischiopubic synchondrosis in symptomatic children. PMID- 7487754 TI - MRI of intraspinal nerve sheath tumours presenting with sciatica. AB - The magnetic resonance imaging (MRI) characteristics of 14 intraspinal nerve sheath tumours (NST) presenting with sciatica were reviewed. The group comprised seven schwannomas, six neurofibromas and one perineuroma. The tumours were either iso- or hypointense with respect to spinal cord on T1-weighted (T1W) images; almost all tumours were hyperintense compared with spinal cord on T2-weighted (T2W) images. The tumours were all detectable on unenhanced T1W images. Nine NST were scanned following Gadolinium-Diethylenetriamine penta acetic acid (DTPA) injection and all showed intense enhancement. This aids differentiation from sequestrated disc fragments. Tumours were more likely to show homogeneous enhancement unless they were recurrent tumours. Rim enhancement occurs more commonly in schwannomas and this can be used to differentiate these from neurofibromas. However, on unenhanced images, schwannomas cannot be distinguished from neurofibromas. Four tumours occurred at T11-T12. There was poor correlation of the site of the lesion with the clinical findings. MRI studies in patients with sciatica should include the lower thoracic region especially if no protruded disc was found in the lumbar region. PMID- 7487755 TI - Clinical follow-up of patients with percutaneously inserted inferior vena caval filters. AB - This is a retrospective clinical follow-up study of 45 consecutive patients who had percutaneous placement of inferior venal caval (IVC) filters in the period 1987 to 1993 to prevent mortality, or severe morbidity, from pulmonary embolism (PE). Ten patients had Gunther filters (GF) and 35 had bird's nest filters (BNF) from 1989. Clinical indications for filter placement were: 23 patients with anticoagulation contraindications, six with anticoagulation failure and recurrent PE, 10 with anticoagulation complications, two with critical PE and four with leg thrombi and PE. Two patients died from recurrent PE after filter placement, one with a GF migrating after replacement, and one with a BNF. Two patients died from severe PE, occurring prior to filter placement. Fifteen patients died in the study period, none due to recurrent PE. Twenty-five patients remain alive, with a mean follow-up period of 21 months, with none having recurring PE. One patient was lost to clinical follow-up. One patient suffered technical difficulty in BNF placement, and underwent surgical removal of a hook penetrating an iliac vein wall. Inferior vena cava filters are efficacious in preventing potentially fatal pulmonary embolism, in both the acute period and the medium term. There are low rates of morbidity associated with their use. PMID- 7487756 TI - Radiological abnormalities in children born as a result of assisted conception. AB - Over 15,000 children are born each year as a result of the various techniques of assisted conception. In 1991, there were 2083 live births in Australia and New Zealand as a result of these techniques. Radiological abnormalities relating to the increased rate of prematurity, a probable increase in the risk of congenital malformations and a number of cases of neural crest tumours seen in these infants will be presented. PMID- 7487757 TI - Colour-coded duplex ultrasonography in selection of patients for transluminal angioplasty. AB - Transluminal angioplasty can be used to relieve symptomatic lower extremity arterial disease with minimal morbidity. This prospective study compares the accuracy of colour-coded duplex ultrasonography with intra-arterial digital subtraction angiography in selection of femoro-popliteal lesions for angioplasty. Isolated, short (< 6 cm) and haemodynamically significant stenosis (> 50% reduction in diameter) were considered for angioplasty. Colour-duplex selection and exclusion was in agreement with angiography in 68 out of 86 lower limbs (79%). The sensitivity, specificity, positive and negative predictive value of colour duplex compared with angiography yielded values of 60.9, 87.5, 60.9 and 85.7%, respectively. The discrepancy between colour duplex and digital subtraction angiography can be partly explained by the tendency of duplex to underestimate the severity and length of disease. It is concluded that colour coded duplex can be used as a screening tool for lower extremity arterial disease to select appropriate lesions for endovascular procedures. The accuracy of duplex in the present study was not perfect and the decision for further intervention should be based on clinical grounds. PMID- 7487758 TI - Orbital hydatid cysts: sonographic and CT appearance. AB - Two cases of unilateral proptosis due to orbital hydatid cysts were evaluated by ultrasound and computed tomography. Image morphology of this rare disease is presented. PMID- 7487759 TI - Imaging features of dissecting neuropathic joints. AB - The imaging appearances of three patients with variable clinical presentations of dissecting neuropathic joints are described. Plain radiographs demonstrated bony debris within the soft tissues adjacent to and distant from neuropathic joints. Computed tomography combined with arthrography confirmed that the soft tissue bony debris communicated with the joints. PMID- 7487760 TI - Ultrasound screening of families with abdominal aortic aneurysm. AB - Recent observations suggest that first degree relatives of patients with an abdominal aortic aneurysm (AAA) are also at risk for the development of this disorder. The first degree relatives of 130 adults with known AAA were invited to attend for an ultrasound examination. Fifty-two eligible relatives (mean age 53 years, range 41-73) from 38 families underwent sonographic screening. Using standard ultrasound criteria no participants were identified with an asymptomatic AAA. Thus, ultrasound of families of patients with AAA has a low yield and may not be useful for screening purposes. PMID- 7487761 TI - Embolization of hepatic artery pseudo-aneurysm facilitated by selective injection of thrombin and thrombin-soaked coils. AB - A case of recurrent haemobilia due to a pseudo-aneurysm of the right hepatic artery is presented. The patient required long-term anticoagulant therapy and, in order to achieve rapid haemostasis, thrombin was used as an adjunct to coil embolization. Rapid thrombosis of the pseudo-aneurysm occurred after selective injection of 200 units of thrombin, and thrombin-soaked coils were employed to occlude the arterial supply. PMID- 7487762 TI - Imaging patterns in melioidosis. AB - Melioidosis is an infectious disease caused by Pseudomonas pseudomallei. It is seldom diagnosed promptly and, if untreated, can lead to an 80-100% mortality rate. Twenty-eight patients with melioidosis were identified over a 6 year period, and their imaging patterns were analysed. Respiratory infections were the commonest form of presentation, frequently shown as diffuse airspace consolidation, and accounted for the highest mortality. Visceral and musculoskeletal infections were associated with chronicity and a high relapse rate. Multifocal splenic abscesses were a common occurrence. Septic arthritis of the knee was frequently seen. The majority of patients had diabetes mellitus and chronic ill-health. An increased awareness of the disease can contribute to its early detection and appropriate treatment. PMID- 7487763 TI - Hypofractionated irradiation for inoperable non-small cell lung cancer. AB - A policy of palliative intent thoracic irradiation was prospectively evaluated in 38 consecutive patients referred for treatment of inoperable non-small cell lung cancer at a single institution. A target dose of 1700 cGy in two fractions 1 week apart was delivered. Characteristics of the treatment group revealed most (87%; 33/38) to be of good-excellent performance status with minimal weight loss before irradiation. Although three patients (8%) had initial metastatic disease, all had symptoms referable to the thorax with cough (71%), dyspnoea (55%), haemoptysis (39%), and chest wall pain (34%) being dominant. Following treatment, the relative risk of maintaining complete response with regard to each of these symptoms was 0.91, 0.40, 0.92 and 0.78, respectively. Overall 70% of patients maintained complete symptomatic response to time of death or last review. Uncorrected median survival was 35 weeks and was comparable to best international end-results for either palliative intent or curative intent radiation schedules. We conclude that the radiation regimen employed is safe, efficacious and eminently resource conscious. Recognition of patient groups who overwhelmingly derive no benefit from conventional fractionation schedules will streamline access to radiotherapy services of patients suitable for radical treatment. PMID- 7487764 TI - Breast cancer following treatment for Hodgkin's disease: the need for screening in a young population. Australasian Radiation Oncology Lymphoma Group (AROLG) AB - Several recent publications have highlighted the issue of an increased risk of breast cancer in women treated with radiotherapy, chemotherapy, or combined modality therapy for Hodgkin's disease. The risk is greatest in women 30 years or younger at the time of treatment. In the Australasian Radiation Oncology Lymphoma Group database, 60% of women fell into this age category. This article reviews the available data pertaining to induction of breast cancer by radiotherapy for Hodgkin's disease. Breast examination should now be an integral part of the long term follow up for these women. There is also a case for the use of screening mammography. Any breast mass developing subsequent to treatment for Hodgkin's disease should be regarded with a high index of clinical suspicion and, accordingly, biopsies should be performed in the majority of cases, even when mammography is negative. PMID- 7487765 TI - Treatment of cutaneous angiosarcomas of the head and neck. AB - A total of six patients with cutaneous angiosarcomas involving the head and neck were treated during the period 1985-1994. The distribution of sites between the scalp and the rest of the head and neck was equal. Initial local treatment involved surgery for four patients and radiotherapy for two patients. In the surgical group, following seemingly adequate macroscopic surgical excision, all four patients had inadequate surgical margins on histology. Five of six patients had disease recurrence within the head and neck and of these a further three went on to have a second episode of recurrence. Median time to recurrence was 8 months. All cases were misdiagnosed clinically at initial presentation and three of six had initial inconclusive histology. Cutaneous angiosarcomas of the head and neck is an aggressive multicentric cancer. Radiotherapy offers the only real chance of obtaining adequate local control even if surgical excision is carried out. Radiotherapy must encompass wide fields and whole scalp irradiation is mandatory for scalp lesions. A combination of laterally opposed photon and electron fields provides a simple, quick and effective means to deliver a tumoricidal dose of 50 Gy to the whole scalp with minimal irradiation of the underlying brain. PMID- 7487766 TI - Dutch variant of Bellini metaphyseal dysplasia: report of two siblings. AB - Two sibling girls with cone-shaped knee epiphyses and metaphyses are described. Bone dysplasia with this rare, distinctive, radiographic finding, was first reported by Bellini and Bardare with only few cases reported thereafter. Velores et al. divided bone dysplasias with cone-shaped epiphyses and metaphyses of the knee in two entities which they named trichoscyphodysplasia and metaphyseal acroscyphodysplasia. Although the authors agree that there is more than one bone dysplasia that presents with these distinctive radiographic knee appearances, they consider that two few cases have been reported to satisfactorily classify this group of disorders. PMID- 7487767 TI - The failed thoracocentesis. AB - How often do we consider a differential diagnosis of diaphragmatic hernia in cases of failed thoracocentesis? How many of us remember that omental fat in the chest can behave in a fluid manner, layering along the chest wall, resulting in the misinterpretation of a decubitus view. A case is presented of a diaphragmatic hernia that was missed on conventional chest radiographs resulting in CT having to be performed. A high degree of suspicion is mandatory to reach the final diagnosis. Radiographs could be used more effectively and CT avoided. PMID- 7487768 TI - Ultrasound and computed tomography appearances of primary hepatic leiomyosarcoma. AB - Primary hepatic leiomyosarcoma is a rare condition. The ultrasound and computed tomography findings in a 73 year old female patient are described. Previous reports have concentrated on the histopathological features and clinical presentation of the condition. PMID- 7487769 TI - Formation and resolution of bilateral traumatic pulmonary pseudocysts. AB - A case of bilateral traumatic pulmonary pseudocysts, an uncommon complication of blunt chest trauma, is presented. The radiologic changes were followed over 6 months until spontaneous resolution had occurred. The mechanism of formation of traumatic pseudocysts and the diagnostic implications are discussed. PMID- 7487770 TI - Hepatic focal nodular hyperplasia: colour Doppler ultrasound can be diagnostic. AB - Focal nodular hyperplasia (FNH) is a rare, benign, hepatic mass lesion that is usually found in young adult females. It is sometimes an incidental finding on abdominal sonography or computed tomography. Its appearance on these studies is not characteristic however, thereby creating a diagnostic problem. This report is of a case which showed a centrifugal, spokewheel pattern of pulsatile blood supply on colour Doppler US. This finding, we suggest, is sufficiently diagnostic to avoid the need for further investigation in some patients, and to direct further imaging more appropriately in others. PMID- 7487772 TI - Haematopoietic marrow hyperplasia simulating transarticular skip metastasis in osteosarcoma. PMID- 7487771 TI - Systemic lupus erythematosus serositis. AB - The imaging appearances of a case of systemic lupus erythematosus, which manifested initially as a serositis, is described. Barium small bowel study showed segments of spiculation with tethering, angulation, and obstruction. On computed tomography there was ascites and segments of asymmetrical thickening of small bowel wall were observed. Laparotomy revealed extensive patchy serosal and peritoneal plaques but biopsy of these lesions did not lead to a definitive diagnosis. The diagnosis was made on the basis of marked elevation of antinuclear and anti-double stranded DNA antibodies. PMID- 7487773 TI - Obturator hernia: diagnosis through medical imaging. AB - A rare case of obturator hernia is presented in a patient with persistent small bowel obstruction. There was a paucity of specific signs and poor general health which caused a delay in diagnosis. Ultimately delayed films from a barium follow through showed barium-filled bowel in the region of the right obturator foramen: A computed tomography scan then confirmed the diagnosis by demonstrating the barium-filled small bowel herniating through the right obturator foramen. Other imaging modalities such as herniography, barium enemas and ultrasound have also been successfully employed to diagnose obturator hernias. PMID- 7487774 TI - The clinical and financial impact of non-invasive vascular testing in the USA. AB - Health care costs in the USA have increased dramatically during the past 10 years, and it is widely believed that they have reached crisis dimensions. Vascular laboratories are a segment of the USA health care industry that has experienced particularly rapid growth in the last decade. In 1992, USA Medicare payments (for patients > or = 65 years) totalled US$304,492,588 for 3,673,695 non invasive vascular studies. This figure represents only one segment of USA health care costs and, if all segments were considered, the 1992 total for vascular laboratory services would probably approach one billion USA dollars. The expansion of vascular laboratory utilization is attributed to a number of factors: (i) increased recognition of the clinical value of non-invasive vascular studies; (ii) replacement of invasive (angiographic) procedures; (iii) expanded surveillance application; (iv) the verification of carotid endarterectomy for treatment of carotid stenosis; (v) widespread, unlimited access; (vi) over utilization; (vii) greed. Widespread awareness of the crisis in USA health care funding has imposed cost-containment pressure where virtually none existed previously. The vascular laboratory is no exception to this trend, and the following measures have been suggested for controlling vascular laboratory costs: (i) demonstration of the clinical and cost effectiveness of non-invasive vascular studies; (ii) utilization review; (iii) pre-approval of requests for vascular studies; (iv) linkage of vascular study reimbursement with clinical diagnosis; (v) limitations on self-referral; (vi) reduced reimbursement; (vii) capitation; (viii) diagnosis-related reimbursement; (ix) accreditation. This article summarizes the factors that have led to increased utilization of vascular laboratory services, and discusses methods proposed for containing vascular laboratory expenditure. PMID- 7487776 TI - Foreign body removal with fine-nosed forceps and assistance of image intensification. AB - Foreign body removal has been developed as a routine radiology department service. Techniques, indications and hazards are described. PMID- 7487775 TI - Lightweight suction drainage-feeding system for oesophagogastric anastomotic leaks. PMID- 7487777 TI - The 1994 Australian Radiology Workforce Committee Report. AB - The task assigned to the College Workforce Subcommittee by the RACR Federal Council is to define current radiologist and trainee numbers, analyse professional demographics, and attempt to forecast radiologist numbers and the medical demand for radiological services. The period of this forecast is for 7 years leading up to 2001. This time course has been chosen on the basis of a 2 year period before any recommendation by Council could be instituted by the Educational Board and 5 years before any change in trainee numbers would be reflected in radiologist numbers. By necessity, assumptions have been made in formulating the final calculations and recommendations; the most difficult being the designation of an 'optimum' number of radiologists for our population. This assignment is an ongoing review and prediction process. The results and conclusions should be compared and contrasted to previous reports, in particular the 1992 Manpower Study. PMID- 7487778 TI - Medical imaging and the Internet. PMID- 7487779 TI - The use of multiple psychotropic medication in the treatment of mental disorders. AB - The concurrent use of multiple psychotropic medication has been reported to be widely prevalent in the treatment of mental disorders, and is considered inferior practice [1]. However, the complexity of the issues demands reappraisal. This paper attempts to discuss the conceptual and practical issues involved. PMID- 7487780 TI - To tell or not to tell: the psychiatrist and child abuse. PMID- 7487781 TI - Shrinking the Kimberley: remote area psychiatry in Australia. AB - The Kimberley, the northernmost region of Western Australia, is still largely an empty wilderness. About twice the size of Victoria, it has a permanent population of some 22,000. Since 1987, the Health Department of WA has provided a psychiatric service to the region, but the practice of psychiatry in such a vast and remote area has proven to be very different from that in larger centres. This paper summarises three years' clinical experience in the region. PMID- 7487782 TI - A survey of psychiatric services for elderly people in Australia. AB - In 1992 a postal questionnaire identified 107 psychiatrists in Australia who, for a significant proportion of their time, were providing psychiatric services to elderly people. Only 18% were female. There were 34 psychiatrists working full time in psychiatry of old age in the public health system. Twenty-three multidisciplinary catchment area psychiatric services for elderly people were identified, but staff to population ratios varied considerably. The ratio of psychiatrists to elderly was 1:30,000--similar to the ratio in a large part of Britain seven years previously. Psychogeriatricians are involved in a broad range of teaching and research activities. The mean number of non-medical staff in a catchment area psychogeriatric service should be increased. PMID- 7487783 TI - Queensland's geographical maldistribution of psychiatrists and Queensland Health's response. AB - There is a major geographical maldistribution and shortage of psychiatrists throughout Queensland. In June 1992, 48 psychiatrists worked outside of Brisbane where 66% of the population reside, while 172 psychiatrists worked in the metropolitan area. The reasons for and the effects of the maldistribution are discussed. Since 1991, Queensland Health has developed a number of key strategies to attract and retain psychiatrists in our non-metropolitan areas. These strategies are outlined. Between July 1992 and December 1993, 14 new full time psychiatrists were recruited. The results of these strategies are encouraging. PMID- 7487784 TI - The effect on hospital admissions of psychiatric case management involving general practitioners: preliminary results. AB - A two year follow-up of two matched groups of subjects with chronic severe mental illness was performed in order to evaluate a new psychiatric case management system. One group (n = 59) received care through psychiatric case management, using an assertive community treatment model that directly involved general practitioners. The other group, matched for age, sex, diagnostic group and number of hospital admissions, received standard outpatient care. Comparing the two years before and after case management, the experimental group showed a dramatic fall in inpatient admission days while the control group admission days remained the same (median difference in admission days across matched subject pairs = 64.5, 95% C.I. from 134.5 to 16). The experimental group remained out of hospital longer before first readmission (Kaplan-Meier survival analysis, P = 0.002). This type of case management programme can shorten or prevent admissions to psychiatric hospitals of patients with chronic mental illness, and increase their time before readmission. PMID- 7487785 TI - Approaches to delivering child and adolescent mental health services: the South Australian experience. AB - Approximately 10% of children and adolescents experience mental health programs, however only a small proportion receive specialised help. Identifying approaches which can provide a balanced and effective service for the large number of children and adolescents with problems is currently a major challenge for child and adolescent mental health services in Australia. In South Australia, following a review in 1983, child and adolescent services were reorganised into two separate but closely related services. This paper draws on experience in South Australia over the last decade to identify approaches which can be employed in six key areas that significantly influence the effectiveness of child and adolescent mental health services. The paper also describes the specific features which were included in the South Australian child and adolescent mental health service to address these issues. PMID- 7487786 TI - Acute psychiatric beds: distribution and staffing in NSW and ACT. AB - This study examined the availability and staffing of acute psychiatry beds in NSW and ACT. "Gazetted" acute psychiatry hospitals (which take compulsory admissions under mental health law) were polled directly for bed numbers, occupancy and staffing for the year 1990-1991. The NSW Department of Health provided beds numbers for non-gazetted and private hospitals. Four analyses sequentially reallocated beds according to the origin of patients to estimate acute bed availability and use by regional populations. Socio-demographic determinants of acute admission rates were measured. Acute "gazetted" beds averaged 13.2 per 100,000 population but ranged from 6.9 to 49.1 per 100,000 when cross-regional flows were considered. "Non-gazetted" beds raised the provision to 15.5 per 100,000 and private beds raised provision further to 24.5 per 100,000. Inner metropolitan provision was higher than rural or provincial provision. The only determinant of the admission rate to gazetted beds was the number of available beds. Bed availability did not affect either bed occupancy or referral of patients to remote hospitals. Nursing staffing of gazetted units was reasonably uniform, although smaller units had significantly more nurses per bed. Medical staffing was highly variable and appears determined by staff availability. The average provision of acute psychiatric beds approximates lowest levels seen in international models for psychiatric services. Average occupancy rates suggest that there is not an overall shortfall of acute psychiatric beds, but uneven bed distribution creates barriers to access. Referral of patients to remote hospitals is not related to actual bed provision in the regions, but appears to reflect attitudes to ensuring local care. Recommendations about current de facto standards are made. Current average nursing and medical staffing standards are reported. PMID- 7487787 TI - Drug use, binge drinking and attempted suicide among homeless and potentially homeless youth. AB - In order to assess the need for drug-related services for at-risk youth, a survey was conducted among young people aged 12-17 years who, owing to severe family discord, were currently living away from home (homeless) or had experienced periods away from home in the past 12 months (potentially homeless). Prevalence of use and of potentially harmful levels of use of alcohol and other licit and illicit drugs were higher than in a comparative population. Of the 155 people interviewed, 54% reported past physical abuse, 28% reported past sexual abuse, and 73% had a family alcohol or other drug history. Of the total, 62% had been in a youth refuge at some time in the past 12 months. Twenty four per cent had been to hospital as a result of alcohol or other drug use and 45% had attempted suicide. Female sex and an interaction between sexual abuse and binge drinking predicted suicide attempts. This study points to the need for a comprehensive approach to interventions for troubled youth which gives greater recognition to mental health issues related to family circumstances, including abuse. PMID- 7487788 TI - Illness aetiology constructs, health status and use of health services among Cambodians in New Zealand. AB - OBJECTIVE: The objectives of this study were to determine, among all adult Cambodians living in Dunedin: prevalence of illness aetiology beliefs; psychiatric and physical health status; pattern of use of health services; relationships between use of health services and demographic factors, illness aetiology constructs and health status; and problems encountered and improvements desired in the local health services. METHOD: 223 (i.e. 93.3% of all) adult Cambodians living in Dunedin were assessed, using a structured interview, in relation to their sociodemographic status, illness aetiology beliefs, physical health status and use of health services. The 28-item version of the General Health Questionnaire was used to assess psychiatric status. RESULTS: Subjects held multiple indigenous and Western illness aetiology constructs. Psychiatric morbidity using the 28item of the General Health Questionnaire (GHQ28) cutoff of 3/4 was 15.7%; despite this only six subjects had ever used specialist psychiatric services. Malaria, intestinal parasitic infestations and heart conditions were the three most frequently reported physical problems. Most subjects had used traditional services in Cambodia but very few had used them in New Zealand. Health service was related to duration of stay in New Zealand. Socio economic status, both physical and psychiatric health status and some illness aetiology constructs. One hundred and forty-two (63.7%) subjects reported problems with use of health services in Dunedin. CONCLUSION: Despite methodological limitations, some useful preliminary data on factors pertaining to use of and satisfaction with health services among Cambodians were collected. Future research should examine family characteristics and the decision-making processes that determine service use. PMID- 7487789 TI - Validation of an Italian translation of the 30-item General Health Questionnaire for use in Australia. AB - A validity study of an Italian translation of the GHQ-30 was conducted on a sample of 96 Italian-born subjects in the community in Perth, Western Australia, using the shortened version of the Present State Examination as the gold standard for a "case" of non-psychotic psychiatric morbidity. The 96 subjects were a subsample of a study of 1109 subjects seen in a community survey, in which 45.8% had a score above 4 on the GHQ-30. It was found that 5/6 was the most appropriate cut-off point. At this level the sensitivity was 81.2%, the specificity 79.7%, the positive predictive value 66.7%, the negative predictive value 89.5% and the overall misclassification rate 19.8%. Adjusting the results to truly reflect the lower case-prevalence rate in the community, using a method suggested by Goldberg, the corresponding values were 57.7%, 92.6%, 66.7%, 89.5% and 14.6%. It was concluded that this version of the GHQ-30 is suitable for use in the Italian born, in the community and in general practice in Australia, using a cut-off point of 5/6. PMID- 7487790 TI - Type A behaviour in the Australian working population. AB - The Type A behaviour pattern (TABP) has been widely researched in Australian studies and, in view of its increasing prominence beyond its original association with risk of coronary heart disease, is likely to be as widely researched in this country well into the future. The Jenkins Activity Survey (JAS), being the most comprehensive self-report instrument used to assess the TABP, appears to be the instrument of choice for the measurement of the TABP in Australia, particularly in epidemiological studies where large samples are involved, or in other studies where either the interview method is impractical or the researchers are untrained in its application. This paper presents normative data on the use of the JAS in the Australian context, derived from a series of empirical studies undertaken by the authors over the past decade. Sociodemographic correlates of these data are presented, and refinements of the JAS for future use are suggested. PMID- 7487791 TI - Sissiness, tomboyism, sex-role, sex identity and orientation. AB - Masculinity and femininity have been studied by self-ratings in independent areas of research: one investigating personality traits considered masculine (M) or feminine (F); the other, behaviours statistically more common in one than in the other sex (sex-linked behaviours). The two approaches were compared for the first time in the present study of 66 male and 51 female medical students. Consistent with previous findings using the second approach, male but not female subjects' opposite sex-linked "sissy" and "tomboyish" behaviours correlated significantly with their reported ratio of homosexual to heterosexual feelings (Ho/Het). Ho/Het did not correlate with either sex's M and F scores, but high M scores in women correlated strongly with several "tomboyish" behaviours. As "tomboyish" behaviours are shown more strongly by women exposed prenatally to increased levels of opposite sex hormones compared to controls, the findings have implications for the biological theory attributing Ho/Het to such prenatal hormonal exposure. PMID- 7487792 TI - Does nursing theory adequately prepare psychiatric nurses to work with delusional patients? AB - The theory and practice of psychiatric nursing has been subjected to little serious research; consequently some nursing interventions recommended by nursing texts lack formal evaluation. This study examined one aspect of practice: the way in which nurses work with delusional patients. Registered psychiatric nurses were asked to describe the way in which they deal with patients whose delusions are a major clinical problem. The findings highlight a disparity between practice and theory. PMID- 7487793 TI - Investigating discharge destinations for schizophrenic patients. AB - An audit of hospital records of 542 patients diagnosed as having schizophrenia discharged in the years 1986-88 from the Princess Alexandra Hospital, Brisbane, showed that 46% of patients left hospital to live with other family members, usually parents or spouses. Analysis of destination suggested that the extent of patient-family connectedness increases when the patient is younger, has had fewer admissions, or has had the illness for a shorter period of time. PMID- 7487794 TI - Neuropsychological outcome from psychosurgery for obsessive-compulsive disorder. AB - Seventeen obsessive-compulsive disorder patients treated with psychosurgery were administered a comprehensive neuropsychological test battery. Their performance on neuropsychological testing was compared with that of an age and severity matched sample of 17 OCD sufferers who had not received psychosurgery. The psychosurgery and control groups did not differ in intellectual or memory functioning, consistent with earlier findings that psychosurgery does not reduce global ability estimates. The psychosurgery group performed more poorly than the control group on an adaptation of the Wisconsin Card Sorting Test, demonstrating the possible impact of frontal lobe lesions on the abilities underpinning the formation and shifting of response sets. PMID- 7487796 TI - Effect of recommended dosage range on the prescribing of antidepressants. AB - The aim of the study was to explore the role that suggested dosage range may play in suboptimal prescribing practice of tricyclic antidepressants. Clinicians practising in adult psychiatry were shown a clinical vignette of melancholia (as per DSM-III-R). One of two questionnaires on treatment practices, specifying either 75-200 mg or 75-425 mg of an unnamed tricyclic antidepressant, were randomly distributed to subjects. A highly significant (p = 0.001) difference was found: those who were given a narrow dosage range prescribed an average of 82.5 mg/day less antidepressant than those for whom the dosage range had been widened, indicating that the range within which dosages are recommended affects prescribing practices. When erroneously constrained, the range may contribute toward sub-optimal prescribing practices and as a consequence toward treatment resistant depression. PMID- 7487797 TI - Glimpses of a Cassel Hospital outpatient group. AB - The following is a selective account of the experience of conducting a weekly out patient analytic group at the Cassel Hospital, London. Certain features of this experience are highlighted: in particular, the significance and value of a male and female co-therapy pair as conductors. Attention is drawn to the fundamentally reparative potential for damaged individuals in group psychotherapy. Such an experience may be valuable, especially in terms of it facilitating a greater degree of personality integration. Certain suggestions are made as to the prerequisites for such a psychotherapeutic approach. The observations may be useful in the provision of psychotherapy services in the Australian health care system. PMID- 7487795 TI - Comparison of propofol and methohexitone in anaesthesia for ECT: effect on seizure duration and outcome. AB - The effect of the induction agents propofol and methohexitone on seizure duration and clinical outcome was assessed in a prospective, randomised, double blind study in 32 depressed patients receiving electroconvulsive therapy (ECT). Assessments comprised Hamilton rating scales pre-ECT, weekly during the course, and at one month follow-up. Patients showed a significant improvement in depression as measured by the Hamilton rating scale across the course of the ECT treatment. This improvement was not significantly different in the propofol and methohexitone groups. Nor was the amount of improvement related to either the total duration of seizure experienced by the patient during the ECT series or the use of tricyclic antidepressants by some patients. PMID- 7487799 TI - Pseudologia fantastica and gender identity disturbance in a Chinese male. AB - A Chinese patient with pathological lying (pseudologia fantastica) and gender identity disturbance is described. The stories "save face" and lack a truly self aggrandising quality. The importance of keeping "face" and maintaining modest behaviour in the Chinese culture is central to the psychopathology. PMID- 7487798 TI - A mass killing in New Zealand. AB - The circumstances surrounding a mass killing in New Zealand are described in conjunction with a review of a number of other similar multiple victim homicides. Due to the rare and extreme nature of such events, it is argued that they should be managed as human disasters by the professionals involved and that stress debriefing should be available for all potential primary, secondary and tertiary victims. PMID- 7487800 TI - Catatonia LSD induced? AB - The case of a patient who developed catatonia one week following Lysergic Acid Diethylamide (LSD) ingestion is presented. The psychosis developed two days after the intake. The catatonic syndrome resolved dramatically following one treatment of electroconvulsive therapy (ECT). This is perhaps the first case report of catatonia following the use of LSD. The need for a diagnostic category of organic catatonia is highlighted. PMID- 7487801 TI - Conflicting constructions of aboriginal suicide: a letter in response to Robert Parker. PMID- 7487802 TI - Antidepressants and side effects. PMID- 7487803 TI - The effect of varying time at -Gz on subsequent +Gz physiological tolerance (push pull effect). AB - INTRODUCTION: Previous studies have demonstrated decreased +Gz tolerance when preceded by 0 Gz or -Gz, referred to as the "push-pull effect." The purpose of this experiment was to observe the effect of varying time duration at -Gz on the push-pull effect. METHODS: During single sessions, six subjects (three men, three women) were subjected to five relaxed exposures to +2.25 Gz on the NAMRL Coriolis Acceleration Platform (CAP). The first and last exposures were control runs that were preceded by +1 Gz. Each experimental run was preceded by -2 Gz for 2, 5, or 15 s. Blood pressure (BP) was monitored using the Finapres at the level of the clavicle. Visual light loss was assessed at +2.25 Gz using a light bar. RESULTS: Mean BP was significantly reduced when the +2.25 Gz exposures were preceded by -2 Gz. Following 15 s of -2 Gz, mean BP decreased more and was slower to recover than for 2 and 5 s of -2 Gz. Reported incidents of visual light loss were: 1 following 2 s, 2 following 5 s, and 4 following 15 s at -2 Gz. There were no reports of visual light loss during control runs. CONCLUSION: During relaxed conditions, the push-pull effect is augmented by increasing duration of the preceding -Gz. PMID- 7487804 TI - Head perturbations during walking while viewing a head-fixed target. AB - BACKGROUND: Inexpensive, head-fixed computer displays are now available that subjects can wear during locomotion. HYPOTHESIS: Viewing a head-fixed visual display will change the characteristics of rotational head perturbations during natural walking. METHODS: Using a 3-axis angular rate sensor, we measured head rotations during natural or treadmill walking, in 10 normal subjects and 2 patients with deficient vestibular function, as they attempted to view (A) a stationary target at optical infinity; and (B) a target at a distance of 20 cm rigidly attached to the head. RESULTS: Normal subjects and patients showed no significant change in the predominant frequency of head rotations in any plane (ranging 0.7-5.7 Hz) during the two different viewing tasks (p > 0.1). Mean peak head velocities (ranging 6-36 degrees.s-1) also showed no difference during the two viewing conditions except in the yaw plane, in which values were greater while viewing the near target (p < 0.005). Predominant frequencies of head rotations were similar in the pitch plane during natural or treadmill walking; however, peak velocities of pitch head rotations were substantially greater during natural walking (p < 0.05). One vestibular patient showed modest increases of head velocity during natural walking compared with normal subjects. CONCLUSIONS: Rotational head perturbations that occur during natural walking are largely unaffected when subjects view a head-fixed target. There is need to study how such perturbations, which induce vestibular eye movements, affect vision of head-fixed displays. PMID- 7487807 TI - Continuous and intermittent personal microclimate cooling strategies. AB - A comparison was made between two personal auxiliary cooling approaches for the relief of thermal stress while wearing the standard USAF Chemical Defense Ensemble (CDE). Subjects exercised at approximately 40% VO2max in either warm (28/24/34 degrees C) or hot (38/26/43 degrees C) environmental conditions, (Tdb/Twb/Tbg degrees C, respectively). During each of three trials, four hours of intermittent work (four work/rest cycles) were attempted. Microclimate air cooling was applied in two different fashions and compared with a control trial during which no cooling was received (NC). In one trial, conditioned air cooling (Tin approximately 20 degrees Cdb) was delivered during rest periods only (intermittent cooling, IC), while during the second trial, ambient air cooling was also applied during the work period in addition to the conditioned air delivered during rest periods (continuous cooling, CC). During the warm condition, exposure cycle time was 45 min work and 15 min rest, while under the hot conditions, exposure cycle time was 30 min work and 30 min rest. Both CC and IC trials resulted in significantly extended work times, lower final rectal temperatures, heart rates, and sweat production (SP) than in the NC trial. Additionally, CC results in significantly lower SP, higher % sweat evaporation, and lower ratings of perceived exertion (RPE) and thermal comfort (TC) than IC at both warm and hot temperatures. Moreover, subjects were better able to maintain thermal equilibrium (i.e., cumulative heat balance) over time using CC compared to IC in the warm environment. The physiological significance of these findings, in some cases, was secondary to the improvement in subjective measures of TC and RPE.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7487805 TI - Physical performance and heat tolerance after chronic water loading and heat acclimation. AB - Heat acclimation (HA) and forced water intake (FWI) have both been found to improve the endurance of human subjects working in hot environments. Therefore, we studied the interaction between HA and FWI. Prior to any treatment (control, AI and BI) the subjects (n = 9) underwent a heat tolerance (HT) test. Thereafter, they were divided into two groups. The first (n = 5) were heat-acclimated (AII), underwent a second HT test, doubled their normal daily water intake for 1 week (AIII), and underwent a third HT test; the second group (n = 4) were subjected to the same protocol, except that the FWI came before and during HA (BII). It was found that both regimens (phases AII and BII) significantly increased work duration. Although the results of the two methods were similar, their combination somewhat lengthened work tolerance time (phases AIII, BIII). Maximal oxygen uptake did not change after HA (BII) or FWI (AII), but the maximal values were attained at significantly lower heart rates, both after BII alone or combined with HA (BIII). In an additional experiment, the time needed to "ride" 15 km on a bicycle ergometer was reduced by 10% after FWI as compared to control time. PMID- 7487806 TI - Undetected neuropsychophysiological sequelae of khat chewing in standard aviation medical examination. AB - BACKGROUND: Khat (Catha edulis Forsk) is a plant whose leaves contain cathinone, a natural amphetamine. Its leaves are chewed for their effect of central nervous system stimulation and sympathomimesis. METHOD: We examined 25 regular (daily) khat chewing flight attendants (RC), 39 social (occasional) khat-chewing flight attendants (SC), and 24 non-khat-chewing aircrew members (NC) who presented for FAA medical examinations, using electroencephalographic (EEG) frequency analysis and 4 psychometric tests measuring perceptual-visual memory and decision-speed. RESULTS: Memory function test scores were significantly lower in khat chewers than NC's, and in RC's than SC's. They correlated significantly negatively with both duration and quantity of khat abuse in both khat-chewing groups. While EEG analysis revealed a statistically significant shift towards fast frequency bands in RC's compared to SC's or NC's, it did not significantly correlate with, or influence, any of the tested memory functions. CONCLUSION: The results suggest an adverse effect of khat chewing on perceptual-visual memory and decision-speed regardless of EEG frequency changes, and are discussed in relation to aviation safety and standard FAA Aviation Medical Examination. PMID- 7487808 TI - Effects of inspired O2 and CO2 on ventilatory responses to LBNP-release and acute head-down tilt. AB - Increases in blood flow and CO2 return to the heart and lungs at the onset of exercise have been proposed to initiate reflexive feedback which increases ventilation (VE), via mechanoreceptors in the heart and/or intrapulmonary CO2 flow receptors. Both lower body negative pressure (-40 mm Hg) release (LBNP release) and acute head-down (-30 degrees) tilt (TILT) provide physiological models to focus upon the effects of increased venous return and CO2 flow on VE, without the confounding influence of limb afferents or the descending efferents associated with central command. We examined the ventilatory responses to LBNP release and TILT while inhaling one of four gas mixtures: a) room air (R); b) 95% O2 (O); c) 95% O2, 1.25% CO2 (LC); and d) 95% O2, 2.25% CO2 (HC). Breath-by breath measurements for VE end-tidal CO2 (PETCO2), tidal volume (VT), and breathing frequency (fB) were taken. VE and VT for HC were significantly higher (p < 0.05) than those for R, O, and LC throughout the test session, while fB and PETCO2 were not significantly different among the gas treatments. VE increased (p < 0.05) above resting baseline with LBNP-release and TILT for R, O, LC, and HC primarily through an elevation of fB. Further, the maximal change in VE following LBNP-release or TILT were not different among inhaled gas mixtures. However, area under the VE curve following LBNP-release and TILT was higher for HC compared to the other gas mixtures. We conclude that these results are inconsistent with the theory that carotid bodies are essential in driving VE with these models.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7487809 TI - Preconditioning with sodium deficits to improve orthostatic tolerance in rats. AB - We hypothesized that prior deficits in total body sodium would improve the subsequent ability of non-hypovolemic rats to maintain arterial pressure when subjected to an orthostatic challenge. This hypothesis was based on similarities in the response of neurohumoral cardiovascular control systems to lower-body negative pressure and negative sodium balance. Sodium deficits were induced in male Sprague-Dawley rats for 7-8 d by feeding sodium restricted diets, or by administering furosemide daily. After this, rats were allowed to regain a positive sodium balance for 1 d by increasing dietary intake or withholding furosemide, and receiving additional normal saline intraperitoneally. Rats subjected to these protocols had equal plasma volumes at the time they were anesthetized and evaluated for orthostatic tolerance. Furosemide-treated rats maintained a higher mean arterial pressure (MAP) than controls (70 +/- 10 vs 34 +/- 9 mm Hg) when rotated to a 90 degrees head-up position for 20 s. Rats receiving the lowest dietary sodium maintained the highest MAP (89 +/- 8 mm Hg) when placed at 60 degrees head-up for 5 min. Both before and during the orthostatic challenges, prior furosemide treatment was associated with a higher peripheral resistance, while prior dietary restriction of sodium was associated with a higher cardiac output. We conclude that preconditioning consisting of chronic sodium deficits can improve orthostatic tolerance in this animal model. The hemodynamic data indicate the different modes of preconditioning may have their primary effect on different determinants of orthostatic tolerance. PMID- 7487810 TI - Effects of short-term spaceflight and recombinant human growth hormone (rhGH) on bone growth in young rats. AB - The purpose of this study was to characterize the effects of short-term spaceflight and recombinant human growth hormone (rhGH) on bone growth in rapidly growing male rats. Bone growth was determined by dynamic bone histomorphometry and rhGH was administered using implanted ALZET osmotic pumps. Treatment with rhGH for 5 d had no effect on body weight but a 4-d spaceflight resulted in a small reduction in the rate of weight gain. Neither spaceflight nor rhGH altered bone length or the rate of bone elongation. In contrast, rhGH and spaceflight each resulted in site- and bone-specific increases (periosteal) and decreases (endocortical) in formation of cortical bone. The results demonstrate that: a) inhibition of periosteal bone formation observed during longer spaceflight is not apparent during a 4-d spaceflight; and b) infusion of rhGH stimulates cortical bone formation in weightless as well as in weight bearing animals. PMID- 7487811 TI - Spaceflight results in depressed cancellous bone formation in rat humeri. AB - Two experiments investigating the effects of short-term spaceflight on cancellous bone turnover were carried out: Physiological Systems Experiment-1 (PSE-1) (a 4-d orbital spaceflight) and PSE-2 (a 10-d flight). Cancellous bone area was not significantly altered by spaceflight in the humerus during either flight. The calculated bone formation rate was unchanged during the 4-d flight but decreased during the 10-d flight. The decrease in calculated bone formation during the longer flight was due to a combination of a statistically significant decrease in mineral apposition rate and a nonsignificant decrease in double label perimeter. The dynamic measurements suggest that spaceflight results in decreases in osteoblast number and activity. Further, the decreases in osteoblast perimeter and osteoid perimeter indicate that a 2-d reloading period was insufficient to restore bone formation to normal. Resorption of the preflight fluorochrome label was not influenced by spaceflight, suggesting that there was no net change in bone resorption. This conclusion is supported by the unremarkable effects of spaceflight on osteoclast number and osteoclast perimeter. Our findings indicate that short-term spaceflight results in depressed osteoblast number and/or activity. Continued decreases in bone formation with normal bone resorption could ultimately lead to cancellous osteopenia and reduced bone strength. PMID- 7487812 TI - Factors affecting differences in supine, sitting, and standing heart rate: the Israeli CORDIS Study. AB - BACKGROUND: Measuring postural changes is among the simplest methods to elicit basic cardiovascular responses from patients who should not be subjected to treadmill stress testing. We attempted to clarify the individual factors that may affect these changes in resting heart rate (HR). METHODS: In a cross-sectional study, 6016 employees in Israeli industries were screened during 1985-87 for cardiovascular disease risk factors (The CORDIS Study). Measures of resting HR (supine, sitting, and standing) were obtained from 5428 subjects. The association between the HR differences for different postures and age, sex, height, body mass index, blood pressure level, health related habits, environmental temperature, and time of the day was analyzed. RESULTS: The difference in HR between supine and standing position was found to be greater among women than men (12.7 vs. 11.6%, p < 0.03). It was independently positively associated with height (p < 0.0001), cigarette smoking (p < 0.0001), and coffee drinking (p < 0.001), and inversely associated with age (p < 0.002), blood pressure level (p < 0.0005), and physical activity at work (p < 0.0001). CONCLUSION: The interindividual variability in HR response to different postures is partially explained by individual variables such as age, sex, height, blood pressure level, and health related habits. PMID- 7487813 TI - Comparison of airline passenger oxygen systems. AB - The principal sources of oxygen for inflight passenger use, scheduled and unscheduled, are examined. Present practices of assessment of the passenger's "fitness to fly" are described. Three partner airlines, British Airways, U.S. Air, and Qantas, catering for more than 8000 oxygen requests annually, are compared. Analysis of customer use suggests that medical oxygen requests are frequently not clinically justified. The growth in demand, for both scheduled and unscheduled use of an expensive resource, supports the need for a "recommended best practice" among carriers. Passengers with respiratory disorders who will most benefit from inflight oxygen are vulnerable either to hypoxia or asthma. PMID- 7487814 TI - Inadequacy of visual alarms in helicopter air medical transport. AB - BACKGROUND: Air medical programs use medical equipment primarily designed for hospital and/or ground transport settings. Many of these medical devices are equipped with auditory alarms of malfunction or deteriorating clinical status. The high ambient noise requires visual scanning of medical devices to detect alarm conditions in the helicopter cabin. PURPOSE: To evaluate the adequacy of visual scanning for alarm conditions in the helicopter air medical environment. METHODS: The helicopter transport program used in this study is staffed with two medical crewmembers. Flight nurse response time (RT) to a visual alarm was assessed during 25 air medical patient flights. RT was measured using a battery powered dual timer device with a red LED visual alarm placed in a fixed position among the medical instruments. The device was activated at a random time point unknown to the medical crew during each patient flight. RT was defined as the elapsed time from activation of the alarm until it was physically switched off by the flight nurse. RESULTS: RT was surprisingly lengthy for the study population with a mean RT of 81.2 +/- 78.4 s (95% CI 48.8-113.5 s). The variability of RT was also surprising ranging from 3 s to > 5 min. CONCLUSION: RT to visual alarms in the air medical environment is lengthy and quite variable. Recognition of malfunction of medical equipment or early signs of clinical instability prior to clinical deterioration cannot be assured by visual scanning for alarm conditions. Alternative alarming systems should be considered and investigated for air medical transport. PMID- 7487815 TI - Use of a spreadsheet program for circadian analysis of biological/physiological data. AB - Biological/physiological data sampled over a period of 24 h can be subjected to a mathematical analysis to determine the presence of circadian rhythmicity. Several procedures have been proposed, most being complex. To render such an analysis simpler and easy to use by non-mathematicians, we developed and tested the cosinor technique using a commonly available commercial spreadsheet (Excel). It can be used to analyze equally or unequally time-spaced data over 24 h with missing data, as well as to calculate the significance and the main limit of the resultant circadian rhythm (mesor, amplitude, acrophase and their confidence limits). Examples of its application to hourly samples of plasma cortisol and minute-by-minute rectal temperatures are shown. PMID- 7487817 TI - A 24-year-old female with new onset of shortness of breath and non-productive cough. PMID- 7487816 TI - Ultrashort microwave signals: a didactic discussion. AB - As a consequence of the variation with frequency of the attenuation and phase velocity of electromagnetic waves in tissue, the shape (variation of the electric field with time) of short electromagnetic pulses incident on tissue changes with depth of penetration. We show that a conjecture that such well-known and long understood changes in pulse shape may generate harmful biological effects is not credible. We also consider the suggestion that such pulses may be useful in medical imaging and the mapping of the electrical properties of complex tissues and show that such use is impracticably difficult for fundamental reasons. PMID- 7487818 TI - Amphetamine use. PMID- 7487819 TI - Inverted immersion as a novel gravitoinertial environment. AB - BACKGROUND: Inverted immersion (II) offers a unique opportunity to swing the orientation of the gravity vector 180 degrees from its usual configuration with vestibular end organs. During II, extrathoracic fluid dynamics are identical to those of upright immersion (UI). II exposes individuals to a novel gravitoinertial environment and, therefore, should produce motion sickness (MS). HYPOTHESIS: II is more provocative of signs and symptoms of MS than UI. METHODS: Nine subjects were exposed once to II and UI. Conventional SCUBA gear was worn. In II, the subject wore a wetsuit which provided 5-7 kg force of positive buoyancy and, with no weight belt, caused him to float while inverted against the underside of a platform. An experiment with UI was identical except that a weight belt was worn which provided 5-7 kg force of negative buoyancy, and the subject stood upright against the bottom of the pool. The experiment was terminated after 3 hours or "upon the onset of the first, clear, persistent feeling of nausea", whichever came first. Throughout the experiment the subject rotated through a series of tasks: assembly of a pipe puzzle, performance of a series of head movements, and ambulation. Immediately post-dive, postural stability was assessed with tandem standing with and without eyes closed and with and without the neck extended 45 degrees. A questionnaire regarding susceptibility to motion sickness was completed pre-dive. RESULTS: No subject terminated the test because of MS during UI; seven subjects terminated the test during II (p < 0.025, McNemar's test). Posture was less stable after II than after UI (p < 0.05, sign test). MS questionnaire results did not predict susceptibility to II. CONCLUSION: II is provocative of MS and postural instability. PMID- 7487820 TI - Cardiovascular effects of varying G-suit pressure and coverage during +1 Gz positive pressure breathing. AB - With the continued evolution of anti-G suits, used to counter the cardiovascular dysfunction arising from +1 Gz hypoxia protection positive pressure breathing (PPB), it was hypothesized that full-coverage anti-G-suits would offer equal protection while using lower inflation pressures than the traditional 4:1 ratio. Nine experienced subjects were exposed to 2 min of 70 mm Hg PPB while wearing either the COMBAT EDGE (CE) and Tactical Life Support System (TLSS) garments with the G-suit inflated to 4 x breathing pressure, and the Advanced Tactical Anti-G Suit (ATAGS) at 4, 3, 2, and 1 x the breathing pressure. All subjects were measured with impedance cardiography (IC), and six were measured simultaneously with both IC and the Cardioscint nuclear probe. IC-estimated stroke volume, relative left ventricular (LV) end-diastolic volume, LV ejection fraction, and peak filling rate were depressed most in the CE and ATAGS 1 conditions (p < 0.001). Heart rate and mean arterial blood pressure changes were highest and lowest, respectively, using the CE and ATAGS 1 garments (p < 0.001). There were no differences in these variables between the TLSS and ATAGS 2-4 conditions. Thus, protection against the PPB-induced fall in LV preload and cardiovascular function may still be adequately afforded by lower G-suit inflation pressures when using full-coverage anti-G suits during PPB intended for high altitude protection. PMID- 7487822 TI - Cockpit-cabin crew interaction: satisfaction with communication and information exchange. AB - BACKGROUND: There were 1240 members of flight deck crews (F/D) and cabin crews (C/C) in SAS Norway who responded to a questionnaire that included issues related to communication between crews. The response rate was 84%. HYPOTHESIS: Possible differences between F/D and male and female C/C concerning evaluations of information exchange and communication between crews were examined. METHODS: A multi-faceted questionnaire containing 250 questions concerning organizational and psychosocial issues, safety questions, and subjective health was employed. Regression analyses were applied to examine predictors of satisfaction with information and communication. RESULTS: One half of aircrew members were dissatisfied with information exchange and cooperation between the cockpit and the cabin in general. About 70% were dissatisfied with interaction related to debriefing and stop. Differences between F/D and C/C were significant concerning debriefing. Cabin crews, and especially females, reported being inadequately informed about specific operational procedures and technical matters. Nine of ten female C/C wanted better information about technical aspects of the airplane or flying. Pilots reported inadequate information about how emergency procedures influence C/C and passengers. Between 72% and 94% were satisfied with interpersonal relationships, while 53% expressed satisfaction with supervision and social support. A majority (86%) confirmed that frequent changes between crews were stressful. Regression analyses yielded significant relationships between frequencies of meetings, supervision and support issues, and scheduling schemes and satisfaction with information exchange and cooperation. CONCLUSIONS: Results suggest a strong need to provide more open, continuous and effective communication between the cockpit and the cabin. Several practical suggestions for improvements are discussed. PMID- 7487821 TI - Attitudes toward urinalysis drug testing within a civilian pilot training program. AB - BACKGROUND: This descriptive-correlational study examined civilian student pilots' attitudes toward urinalysis (UA) drug testing and the pilots' opinions regarding effectiveness, adequacy, and fairness of the method as a deterrent for substance abuse among pilots. HYPOTHESIS: There will be significant differences among attitudes of professional aviation student pilots in mandatory and nonmandatory UA drug tested sections of a flight program when considering the pilots' flight hours, ages, gender, anxiety produced by UA drug testing, opinions on drug and alcohol abuse by pilots on the flight schedule, and their attitudes toward the testing process. METHODS: A Likert-scaled questionnaire was completed by the student pilots who were enrolled in mandatory and nonmandatory UA drug tested class sections within a large civilian aviation flight program. RESULTS: Pilots felt mandatory UA testing was less anxiety-producing (t = 2.25, p < 0.05) than the students in nonmandatory tested flight classes. The pilots in nonmandatory tested sections felt more strongly (t = 2.55, p < 0.01) that drug use existed among pilots on the flight schedule than mandatorily tested pilots. Significant intercorrelations (p < 0.05) emerged among the variables age, gender, flying hours, randomization of UA testing, and attitudes toward the UA testing process and its' effectiveness. CONCLUSIONS: Based on the results, mandatory UA drug testing appeared to decrease substance abuse among pilots on the flight schedule. PMID- 7487823 TI - Motion sickness incidence during a round-the-world yacht race. AB - Motion sickness experiences were obtained from participants in a 9 month, round the world yacht race. Race participants completed questionnaires on their motion sickness experience 1 week prior to the start of the race, during the race, and following the race. Yacht headings, sea states, and wind directions were recorded throughout the race. Illness and the occurrence of vomiting were related to the duration at sea and yacht encounter directions relative to the prevailing wind. Individual crewmember characteristics, the use of anti-motion sickness drugs, activity while at sea, and after-effects of yacht motion were also examined with respect to sickness occurrence. Sickness was greatest among females and younger crewmembers, and among crewmembers who used anti-motion sickness drugs. Sickness varied as a function of drug type and activity while at sea. Crewmembers who reported after-effects of yacht motion also reported greater sickness while at sea. The primary determinants of motion sickness were the duration of time spent at sea and yacht encounter direction to the prevailing wind. PMID- 7487824 TI - Speech production and cognitive deficits on Mt. Everest. AB - Deficits in speech motor control and in the comprehension of syntax were observed as five members of the 1993 American Sagarmatha Expedition ascended Mt. Everest. We analyzed speech recordings and cognitive test scores of the climbers at different altitudes. The mean "voice onset time" interval that differentiates "voiced" stop consonants from their "unvoiced" counterparts (e.g., a [b] from a [p]) decreased from 24.0 ms at Base Camp to 5.4 ms at Camp Three. The time needed to comprehend simple spoken English sentences increased by 50% at higher altitudes, and was correlated with speech motor deterioration. This pattern of deficits is similar to that noted for Parkinson's disease and may reflect disruption of subcortical pathways to prefrontal cortex. Similar procedures could be used to remotely assess cognitive impairments caused by hypoxia, carbon monoxide or alcohol intoxication, or drugs, in order to monitor crew behavior in aeronautics and spaceflight operations, or to evaluate the treatment of neurodegenerative diseases such as Parkinson's disease. PMID- 7487825 TI - Comparison of heart rate and arterial pressure spectra during head up tilt and a matched level of LBNP. AB - Lower Body Negative Pressure (LBNP) can be used to stimulate cardiovascular regulation by inducing blood shifts similar to those produced during head up tilt (HUT). It is unclear, however, whether similar blood shifts produced by these two stresses evoke similar cardiovascular regulatory responses. Hence, we compared the autonomic components of cardiovascular responses to 50 degrees HUT and a matched level of LBNP. A level of LBNP that produced changes in calf circumference similar to those produced during the first 3 min of 50 degrees HUT was considered to be a matched level. Autonomic components of cardiovascular responses were determined by spectral analysis of heart rate and blood pressure. Results from nine subjects showed that in terms of changes in calf circumference at the end of 3 min, 50 degrees HUT and 48 mm Hg LBNP were similar (2.13% and 1.94%). During 20-min exposures to HUT and LBNP, the increase in heart rate during LBNP was greater (+7 bpm) than HUT, while blood pressure increases were similar. For heart rate and blood pressure spectra, power in the respiratory frequency region (0.25 Hz) decreased and power in the low frequency region (0.03 Hz) increased similarly during HUT and LBNP. These results indicated that 50 degrees HUT and a matched level of LBNP evoked similar autonomic responses in cardiovascular regulation, with the autonomic balance shifted toward increased sympathetic and decreased parasympathetic influence. PMID- 7487826 TI - Simulated microgravity increases cutaneous blood flow in the head and leg of humans. AB - BACKGROUND: The cutaneous microcirculation vasodilates during acute 6 degrees head-down tilt (HDT, simulated microgravity) relative to upright conditions, more in the lower body than in the upper body. HYPOTHESIS: We expected that relative magnitudes of and differences between upper and lower body cutaneous blood flow elevation would be sustained during initial acclimation to simulated microgravity. METHODS: We measured cutaneous microvascular blood flow with laser Doppler flowmetry at the leg (over the distal tibia) and cheek (over the zygomatic arch) of eight healthy men before, during, and after 24 h of HDT. Results were calculated as a percentage of baseline value (100% measured during pre-tilt upright sitting). RESULTS: Cutaneous blood flow in the cheek increased significantly to 165 +/- 37% (mean +/- SE, p < 0.05) at 9-12 h HDT, then returned to near baseline values by 24 h HDT (114 +/- 29%, NSD), despite increased local arterial pressure. Microvascular flow in the leg remained significantly elevated above baseline throughout 24 h HDT (427 +/- 85% at 3 h HDT and 215 +/- 142% at 24 h HDT, p < 0.05). During the 6-h upright sitting recovery period, cheek and leg blood flow levels returned to near pre-tilt baseline values. CONCLUSIONS: Because hydrostatic effects of HDT increase local arterial pressure at the carotid sinus, baroreflex-mediated withdrawal of sympathetic tone probably contributed to increased microvascular flows at the head and leg during HDT. In the leg, baroreflex effects combined with minimal stimulation of local veno-arteriolar and myogenic autoregulatory vasoconstriction to elicit relatively larger and more sustained increases in cutaneous flow during HDT. In the cheek, delayed myogenic vasoconstriction and/or humoral effects apparently compensated for flow elevation by 24 h of HDT. Therefore, localized vascular adaptations to gravity probably explain differences in acclimation of lower and upper body blood flow to HDT and actual microgravity. PMID- 7487828 TI - Critical incident stress intervention following fatal aircraft mishaps. AB - This article offers guidelines for conducting preventive mental health interventions using a Critical Incident Stress Debriefing (CISD) model with Air Force and other aviation communities that have experienced a fatal aircraft mishap. The guidelines are based on the authors' experiences in providing assistance to several hundred active duty military personnel and their families following two separate mishaps. The article outlines the basics of the CISD model, briefly describes the two mishaps, and then discusses the lessons learned. PMID- 7487827 TI - The role of functional endoscopic sinus surgery in the management of recurrent sinus barotrauma. AB - Sinus barotrauma occurs when an individual is unable to equilibrate the pressure within his sinuses with atmospheric pressure. Aviators affected by recurrent sinus barotrauma are unfit to fly until the underlying cause is established and treated. While most cases result from intranasal pathology, a significant number are the result of sinus pathology or anatomical abnormalities. This latter group have been difficult to manage in the past but the advent of computerized tomography and the Hopkin's rod endoscope have allowed them to be operated on with precision and safety. Complications of this treatment are uncommon and the aviator has usually been able to resume full flying duties after undergoing a decompression test. In our experience it is rare for an aviator who has passed a decompression test to have further episodes of sinus barotrauma. PMID- 7487829 TI - The feasibility of collecting in-flight EEG data from helicopter pilots. AB - An assessment of whether valid EEG data could be collected on helicopter pilots in flight was conducted. Each subject provided eyes-open/eyes-closed EEG in the laboratory and in a helicopter. During flights, EEG's were monitored on the ground in real-time via radio telemetry. Analyses were conducted on the data recorded from Fz, Cz, Pz, P3, P4, O1, and O2 from eight subjects. Delta activity at one recording site (Fz) was higher in the aircraft than in the laboratory probably because of increased eye movements. Both theta and alpha activity at several sites also were increased in the aircraft, and alpha activity at all electrodes showed the expected augmentation from eyes-opened to eyes-closed; however, there were no interactions indicative of problems detecting normal alpha changes due to eye closure in the aircraft. Beta activity recorded from Cz and O1 was elevated during flight testing, but it was concluded that at least the O1 effect was due to muscle artifact in the more active environment. While there were more recording artifacts in the helicopter than in the laboratory, the overall results show it is possible to telemeter EEG from helicopter pilots in flight. Follow-on studies are needed to assess whether recordings can be obtained while pilots are performing flight-related tasks. PMID- 7487830 TI - The respiratory system in a cold environment. AB - Acute or chronic cold exposure elicits several effects on the respiratory system. Pulmonary mechanics are compromised by bronchoconstriction, airway congestion, secretions and decreased mucociliary clearance. These responses are active in cold- or exercise-induced asthma, and are possibly responsible for decreased immune function and protection against airborne pollutants. The primary ventilatory effect of cold air is to decrease baseline ventilation and respiratory chemosensitivity. Although these responses provide significant protection against heat loss in many animals, the effect in humans is minimal. Cold exposure also elicits an increase in pulmonary vascular resistance. This stimulus is synergistic with hypoxia and may mediate pulmonary hypertension and edema at altitude. Chronic exposure to cold environments results in morphological changes such as increased numbers of goblet cells and mucous glands, hypertrophy of airway muscular fascicles and increased muscle layers of terminal arteries and arterioles. These latter two factors may play a role in the symptoms of chronic obstructive pulmonary disease and bronchitis, high altitude pulmonary hypertension and edema, and right heart hypertrophy. PMID- 7487831 TI - Cases from the aerospace medicine residents' teaching file. Case 61. A pilot with vestibular neuritis. AB - A pilot presented with a history of intermittent unilateral tinnitus and transient disequilibrium following a brief viral illness. The clinical course and diagnostic evaluation were consistent with a diagnosis of vestibular neuritis. The clinical presentation, evaluation, and diagnosis of a pilot with vestibular neuritis are discussed. PMID- 7487832 TI - An Afghan airlift. PMID- 7487833 TI - Problems with a large organ. PMID- 7487834 TI - Classification of decompression sickness. PMID- 7487835 TI - Erythropoietin after long-term spaceflight (30 d) PMID- 7487836 TI - Airborne transmission of infectious diseases aboard aircraft. PMID- 7487837 TI - Directory issue of the Aerospace Medical Association. PMID- 7487838 TI - Development and familiality of sexual orientation in females. AB - The development and familial clustering of sexual orientation were studied in 358 heterosexual, bisexual, and homosexual women. Sexual orientation, as measured by the Kinsey scales, was diverse yet showed statistical congruity and stability over a 1- to 1.5-year time span. Developmental patterns, as measured by retrospective reports on the ages of first sexual or romantic attraction and of self-acknowledgement of sexual orientation were very similar in the heterosexual and lesbian subjects except for the difference in object choice. The bisexual subjects displayed intermediate patterns that were more similar to the heterosexuals' on most facets yet closer to the lesbian subjects' on other dimensions. Familial clustering of nonheterosexual orientation was significant. Using two criteria, elevated rates of nonheterosexuality were found in four classes of relatives: sisters, daughters, nieces, and female cousins through a paternal uncle. The current data are not sufficient to distinguish between genetic and shared environmental sources of this familial aggregation. We discuss the possibility of using developmental criteria to differentiate between inherited and cultural sources of variation in female sexual orientation. PMID- 7487839 TI - Heritability of cognitive abilities in adult twins: comparison of Minnesota and Swedish data. AB - Cross-sectional reports suggest heritability of cognitive ability increases throughout adulthood. To investigate this hypothesis, quantitative genetic analyses were conducted on four measures of cognitive ability (verbal, spatial, perceptual speed, memory). Data from Minnesota and Swedish twin studies of aging were compared. Heritability estimates and the factor structure of cognitive abilities could be equated across younger twins (age, 27-50) and middle-aged twins (age, 50-65) from both studies, suggesting stability of heritability during adulthood. The heritability of 81% for a general cognitive factor confirmed earlier findings of high heritability in younger and middle-aged samples. Older Swedish twins (age, 65-85) demonstrated significantly lower heritability estimates for cognitive abilities (54%) and a significantly different factor structure of cognitive ability. PMID- 7487840 TI - A gene-culture model of human handedness. AB - A model of handedness incorporating both genetic and cultural processes is proposed, based on an evolutionary analysis, and maximum-likelihood estimates of its parameters are generated. This model has the characteristics that (i) no genetic variation underlies variation in handedness, and (ii) variation in handedness among humans is the result of a combination of cultural and developmental factors, but (iii) a genetic influence remains since handedness is a facultative trait. The model fits the data from 17 studies of handedness in families and 14 studies of handedness in monozygotic and dizygotic twins. This model has the additional advantages that it can explain why monozygotic and dizygotic twins and siblings have similar concordance rates, and no hypothetical selection regimes are required to explain the persistence of left handedness. PMID- 7487841 TI - Genetic vs. environmental contributions to human handedness: insights gained from studying individuals with unilateral hand injuries. AB - Four hundred eleven individuals who had suffered temporary unilateral hand injuries were assessed on five hand preference behaviors after their recovery. Measures of shifts to contralateral hand use experienced during and after the recovery period were taken. The best predictor of hand use shifts during the injury recovery period was whether the injury was to the preferred or nonpreferred hand. The incidence of postinjury contralateral shifts was low but was greater than that observed over a simulated injury period in a control group of 384 individuals who had not experienced a hand injury. PMID- 7487843 TI - Nonrandom mating in the two-spot ladybird (Adalia bipunctata): the influence of weight on mating success. AB - In some populations of the two-spot ladybird, Adalia bipunctata, there is evidence that melanics of both sexes have a general mating advantage over the typical morph. There is also evidence that some female ladybirds possess a simple mating preference for melanic males. We have determined the influence of body weight on mating success in the two-spot ladybird and assessed whether weight differences might influence the mating success of the melanic morph. We found that the "formal mating tests" used in previous studies of the two-spot ladybird did not detect any influence of weight on mating success. Using more sensitive "singleton" tests, however, a significant mating advantage to heavier males was detected, irrespective of morph. There was also evidence in favor of a similar advantage to heavy females, but this was not present in all ladybird groups that were tested. Heavy individuals neither produced more eggs in matings nor showed higher activity rates. We suggest that some form of mate competition favors large individuals: for example, the larger an individual, the greater is the chance of an encounter with a potential mate. There is no evidence in this study that either sex chooses heavier mates. Effects of weight might contribute to the general melanic mating advantage found in some populations but cannot account for all the data in favor of a female mating preference for melanic males. PMID- 7487842 TI - Intrapair differences in personality and cognitive ability among young monozygotic twins distinguished by chorion type. AB - We evaluated placentation effects on behavioral resemblance of 44 pairs of monozygotic (MZ) twin children. Tested at ages 4-6, the twins' zygosity and placental type had been determined at their delivery. The sample included 23 monochorionic (MC) and 21 dichorionic (DC) MZ twin pairs: DC-MZ twins result from separation of blastomeres within 72 h of ovulation; MC-MZ twins arise from later duplication of the inner cell mass. Twins were individually administered the McCarthy Scales of Cognitive Ability, while their mothers separately rated each cotwin on an individualized 280-item form of the Personality Inventory for Children (PIC). Absolute differences between MC-MZ cotwins were smaller than those between DC-MZ cotwins for all 20 PIC scales, significantly so for 3 of 4 factor scales, 8 of 12 clinical scales, and 2 of 4 validity/screening scales from the PIC; in contrast, no consistent differences in intrapair resemblance of mono- and dichorionic MZ twins were found for the McCarthy Scales. The chorion differences found in the PIC data cannot be due to genetic differences, because all pairs are monozygotes; nor are they associated with differences in parity, gestational age, birth weight, maternal education, palmar dermatoglyphic asymmetry, or maternal knowledge of chorion type. We interpret our findings as suggestive evidence that variation in timing of embryological division, with effects on MZ twins' placental vasculature, has significant consequences for some dimensions of their behavioral development, as well. PMID- 7487845 TI - Potential stock differences in the social behavior of rats in a situation of restricted access to food. AB - The social behavior of outbred Long-Evans (LE) and Wistar (WI) rats was compared in a situation where access to food was particularly difficult (clearing an aquatic barrier, plus the necessity of carrying the food back to the home cage). In groups of either six WI or LE rats, only about 50% of individuals carried the food, and the others survived by attacking those that did. However, behavioral profiles associated with these acts were different in the two cases: LE carriers, contrary to WI carriers, restole some food, and LE noncarriers expressed more agonistic behavior and were more often attacked than were the WI noncarriers. Food flow and all associated, interactive behaviors were more complex in the LE than in the WI rats, indicating the likelihood of potential genetic differences in this testing situation. PMID- 7487844 TI - Congenic AB mice: a novel means for studying the (molecular) genetics of aggression. AB - Congenic strains (CS) of mice were established to identify genomic regions which are associated with the male behavioral trait "isolation-induced aggression" (iia). For this purpose the trait was backcrossed for 10 generations onto the genetic background of a closely related, but nonaggressive, strain. Brother/sister matings were subsequently performed for 10 generations. Genomic screening for "iia-associated" markers was performed via multilocus DNA fingerprinting with a panel of oligonucleotide probes containing simple tandem repetitive motifs. Pools of DNAs from 10 mice each were composed from inbred generations to minimize residual genetic variability in the CS. The representation of iia-associated DNA fingerprint bands was additionally ascertained by investigating the individual mouse genomes constituting the pools. The CS system may allow rational approaches to the behavioral trait "aggression," even under various experimental conditions of different environments. PMID- 7487847 TI - A comparison of two psychological treatments for bulimia nervosa: implications for models of maintenance. AB - In an effort to elucidate the role of cognitive factors in the maintenance of bulimia nervosa, the efficacy of two psychological treatments was examined in a randomised control trial: cognitive behaviour therapy in the absence of explicit exposure instructions was compared with exposure and response prevention treatment in the absence of cognitive restructuring procedures. In the short term both treatments were successful at effecting substantial improvement in both the specific and the non-specific psychopathology of the disorder. However, at a one year follow up, whilst improvements were well maintained for those who had received the cognitive-behavioural treatment, virtually all of those who had responded to the purely behavioural treatment had relapsed. This provides some support for the cognitive model of the maintenance of bulimia nervosa. Nevertheless, the two treatment groups could not be distinguished on post treatment measures of cognitive disturbance and neither was it the case that residual levels of cognitive disturbance, as assessed, predicted relapse. This may suggest that the level at which the necessary cognitive change takes place may not be accessible by conventional assessment procedures. PMID- 7487846 TI - Quantitative trait loci associated with the behavioral response of B x D recombinant inbred mice to restraint stress: a preliminary communication. AB - Quantitative trait loci (QTL) analysis was used to make provisional identification of loci containing genes influencing vulnerability to stress. The effect of restraint stress on open-field activity was measured in C57BL/6J and DBA/2J inbred strains of mice and in 22 B x D recombinant inbred strains of mice. QTL analyses were performed by correlating the behavioral delta scores for each group with the strain distribution pattern of 1300 markers for the B x D mice. A significant association was found between postrestraint rearings during min 5 through 8 in the open field and the Lamb2 marker on chromosome 1 (r = .718, p < .0001). Significant associations at the p < .0001 level were also found between baseline open-field rearings of control mice during min 0 through 5 and the Zp3, Ache, and Mr66-1 markers on chromosome 5, baseline open-field rearings of control mice during min 5 through 8 and the Pmv42 marker on chromosome 15, and open-field rearings of experimental mice during min 0 through 5 and the D11Ncvs61 marker on chromosome 11. PMID- 7487848 TI - Inference processes in obsessive-compulsive disorder: some clinical observations. AB - In this paper we outline a cognitive model of Obsessive-Compulsive Disorder (OCD) which proposes that the core belief of OCD evolves through a series of illogical inferences. These faulty inference processes involve inferring the plausibility of events on the basis of irrelevant associations, dismissing actual evidence on the grounds of going beyond surface reality to a deeper reality, and finally inferring that a completely fictional narrative is a remote probability. A therapy aimed specifically at changing these inference processes is illustrated with case examples of OCD clients who had not benefited from conventional behavior therapy. The inference based approach (IBA) complements existing cognitive-behavioral therapy but suggests that in certain cases, the conventional cognitive therapy view of OCD beliefs as exaggerated fears of remote possibilities may actually reinforce the obsessional belief since even remote events are real. The IBA on the contrary suggests that an important goal in therapy is to highlight this confusion found in OCD between imagination and reality and illuminate for the OCD client how their compulsions, far from reassuring them about a remote possibility, actually take them further away from reality and reinforce their imaginary doubt. PMID- 7487849 TI - The value of possessions in compulsive hoarding: patterns of use and attachment. AB - Hoarding behavior, patterns of use of possessions, and emotional attachment to possessions were examined among a sample of female undergraduates and a sample of community volunteers. Hoarding behavior was associated with a decreased frequency of use of possessions and excessive concern about maintaining control over possessions. Furthermore, high scores on the hoarding scale were associated with higher levels of perceived responsibility for: (1) being prepared; and (2) the well-being of the possession. Hoarding was also associated with greater emotional attachment to possessions and to the reliance on possessions for emotional comfort. The implications of these findings for the definition of hoarding are discussed. PMID- 7487850 TI - Origins of common fears: effects on severity, anxiety responses and memories of onset. AB - The purpose of the present study was twofold: First, to test Rachman's (1976) theory (Behaviour Research and Therapy, 14, 125-131) which predicts that "directly" conditioned fears will differ from "indirectly" conditioned fears in magnitude and anxiety response patterns. Secondly, to explore validity issues related to the questionnaire methodology typically used in fear acquisition research. The questionnaire comprised 3 anxiety scales and 3 fear-onset questions used in prior research, a specifically developed 36-item fear list and 2 validity check items. One hundred and ninety-one Ss completed the questionnaire. After selecting and rank-ordering 10 fears from the fear list, Ss answered questionnaire items for their first- and then their tenth-ranked fear. Results failed to confirm Rachman's predictions: A significantly greater proportion of Ss ascribed highly-feared and moderately-feared situations or things to direct conditioning. In addition, differential anxiety response patterns were not present for different levels of fear. However, results supported the prediction that direct-conditioning ascriptions would be endorsed with greater certainty. The findings suggest that direct-conditioning experiences may be more memorable than indirect-conditioning events. The theoretical and methodological implications of the findings are discussed. It is suggested that future research either employ methodologies more suited to investigating causal relationships or that Rachman's (1976) theory be blended with an attributional account of fear acquisition. PMID- 7487851 TI - "If I feel anxious, there must be danger": ex-consequentia reasoning in inferring danger in anxiety disorders. AB - It has been suggested that neurotic patients engage in 'emotional reasoning', i.e. draw invalid conclusions about a situation on the basis of their subjective emotional response. The present experiment investigated whether anxiety patients infer danger on the basis of their anxious response, whereas normals infer danger only on the basis of objective information. Four groups of anxiety patients (52 spider phobics, 41 panic patients, 38 social phobics, and 31 other anxiety patients) and 24 normal controls made ratings of the danger they perceived in scripts in which information about objective safety vs objective danger, and anxiety response vs non-anxiety response information were systematically varied. As hypothesized, anxiety patients were not only influenced by objective danger information, but also by anxiety response information, whereas normal controls were not. The effect was neither situation-specific, nor specific for panic patients. This tendency to infer danger on the basis of subjective anxiety ('ex consequentia reasoning') may play a role in the development and maintenance of anxiety disorders. PMID- 7487852 TI - A follow-up study of cognitive bias in generalized anxiety disorder. AB - Patients with generalized anxiety disorder (GAD) without concurrent depression (n = 11) and normal controls (n = 17) were tested twice, about 2 months apart, on a modified Stroop colour-naming task, which presented anxiety-related, depression related and neutral words in masked and unmasked exposure conditions. GAD patients received cognitive behaviour therapy in the test-retest interval, and were also retested at follow-up, about 20 months after initial testing. GAD patients showed interference in colour-naming negative words across both masked and unmasked conditions before treatment, but not post-treatment, compared with controls. Reduced interference effects of masked threat words over time correlated with reduced ratings of anxious thoughts at post-treatment, and at follow-up, in GAD patients. Thus, the preconscious bias for threat information in GAD appears to vary over time in association with changes in anxious thoughts and worries. PMID- 7487853 TI - Experimental manipulations of responsibility: an analogue test for models of obsessive-compulsive disorder. AB - Several authors attribute excessive responsibility a predominant role in Obsessive-Compulsive Disorder (OCD) [Salkovskis, P. M. (1985) Behaviour Research and Therapy, 23, 571-583; Rachman, S. (1993) Behaviour Research and Therapy, 31, 149-154; van Oppen, P. & Arntz, A. (1994) Behaviour Research and Therapy, 32, 79 87]. The present studies aimed at demonstrating the link between different levels of perceived responsibility and checking behaviors by experimentally manipulating responsibility in non-clinical Ss. In the first study, a sound recognition task was used to compare checking behaviors in Ss with high (HR) and low (LR) perceived responsibility. Only one variable was significantly different, Ss from the HR group reporting more anxiety during the task than Ss from the LR group. Results did not support a link between responsibility and checking behavior. In a second study HR and LR Ss were compared on a manual classification task. Subjects from the HR group hesitated and checked more, and reported more preoccupation with errors and anxiety during the task than Ss from the LR group. Since perceived severity of the outcome was the most variable affected by the manipulation, the implications for current models of OCD are discussed and an alternative explanation is attempted. Finally, clinical implications are examined and suggestions are made for future directions of research. PMID- 7487854 TI - One-session cognitive treatment of dental phobia: preparing dental phobics for treatment by restructuring negative cognitions. AB - The purpose of this study was to investigate the effectiveness of a single session of cognitive restructuring in a sample of phobic dental patients. Fifty two patients were randomly assigned to one of three conditions: cognitive restructuring (modification of negative cognitions), provision of information (about oral health and dental treatment), and a waiting list control condition. Both interventions maximally lasted one hour. In comparison with the waiting list control condition and the information intervention condition, the cognitive intervention condition not only showed a large decrease in frequency and believability of negative cognitions, but also exhibited a clear decline in dental trait anxiety. Analysis at a follow-up of one year demonstrated a further, drastic reduction in dental anxiety in both intervention conditions, wherein the difference among these conditions was not maintained. It is concluded that it is possible to obtain substantial reductions of dental trait anxiety through a single session of cognitive restructuring. Nevertheless, repeated exposure to the dental situation seems necessary for a further reduction of anxiety. PMID- 7487855 TI - Aetiology of spider phobia: classificatory differences between two origins instruments. AB - The origins of fear of spiders was investigated in 33 spider phobic Ss entering a treatment study. All Ss completed Menzies and Clarke's (1993a) Origins Questionnaire (OQ) (Behaviour Research and Therapy, 31, 355-365) and Ost and Hugdahl's (1981) Phobic Origins Questionnaire (POQ) (Behaviour Research and Therapy, 19, 439-447). The results for the individual questionnaires were similar to those reported in previous studies. However a comparison of assignments of origin category for the two questionnaires showed widely discrepant results. The POQ returned 17 positive responses for classical conditioning, the OQ only 2. By contrast the OQ returned 10 origins as 'non-conditioning traumatic event', 9 of which returned positive responses for classical conditioning on the POQ. A further 15 Ss on the OQ were categorised as 'always been this way', the preponderance of these being classified as vicarious or informational in origin on the POQ. The POQ does not have a specific question or category for non associative acquisition of phobias. The results of the origins assignments are detailed and their relation to the structure and questions of the questionnaires is outlined. Past studies using the POQ have produced results supporting acquisition of a substantial proportion of phobias by classical conditioning. The findings of this study suggest that such results mainly reflect inherent bias in that instrument. PMID- 7487856 TI - One-session therapist directed exposure vs two forms of manual directed self exposure in the treatment of spider phobia. AB - Fifty-two patients with spider phobia, fulfilling the DSM-III-R criteria for simple phobia, were assessed with behavioral, physiological and self-report measures. They were randomly assigned to five different treatment conditions: (1) one session therapist-directed exposure (maximum 3 hours), (2) specific manual based treatment in the home, (3) specific manual-based treatment at the clinic, (4) general manual-based treatment in the home and (5) general manual-based treatment at the clinic. The results show that therapist-directed one-session treatment was significantly more effective than three of the manual-based treatments, both at the post-treatment and follow-up stages. Specific manual based treatment at the clinic was significantly more successful than the other manual-based treatments, but only at follow-up. The proportion of clinically significant improved patients at follow-up was 80% in the therapist-directed group compared to 63% for the specific manual-based treatment at the clinic, 10% for specific manual-based treatment in the home, 9% for general manual-based treatment in the home, and 10% for general manual-based treatment at the clinic. The conclusion that can be drawn is that one-session therapist-directed treatment is the treatment of choice for spider phobia but manual-based treatment is a good alternative in some cases. PMID- 7487858 TI - Psychomotor retardation found in college students seeking counseling. AB - Psychomotor retardation (PMR) has been documented for inpatient populations. Futterman and Tryon (1994) [Journal of Behavior Therapy and Experimental Psychiatry, 25, 41-48] provided the first report of PMR in a female outpatient sample. Because 8 of their 11 depressed Ss were receiving medication, the possibility remains that Futterman and Tryon's findings were due to medication side effects. The present study documents PMR in a nonmedicated male and female college population. Two-week, 24-h per day, content-valid behavioral measurements were obtained using step-counters from 73 college students seeking counseling. The 20 males and 53 females completed the Inventory to Diagnose Depression, the State-Trait Anxiety Inventory, the Marlowe-Crowne Social Desirability Inventory, and a structured interview. Regression analysis revealed that depression was negatively related to daytime activity. Significant age and gender effects were statistically controlled. PMID- 7487857 TI - The assessment of unwanted intrusive thoughts: a review and critique of the literature. AB - In this paper we review the assessment and measurement of normal unwanted intrusive thoughts, images, and impulses that are considered the basis of clinical obsessions. After highlighting some difficulties with how the definition of cognitive intrusion has been applied to the development of assessment measures, we evaluate the construct validity of a number of retrospective self report instruments such as the Intrusive Thoughts Questionnaire, Cognitive Intrusions Questionnaire, and Obsessional Intrusions Inventory, as well as interview and diary procedures. Measures of personal responsibility and meta cognitive beliefs, which are still in the developmental phase, are also discussed. We conclude with a number of recommendations and areas of further research which would strengthen the construct validity of measures of intrusive thoughts and related constructs. PMID- 7487860 TI - Confirmatory factor analysis of The Aggression Questionnaire. AB - A confirmatory factor analysis of the factor structure of The Aggression Questionnaire created by Buss and Perry (1992) [Journal of Personality and Social Psychology, 63, 452-459] was conducted to assess whether the scale's purported 4 factors emerged. The results generally supported the 4-factor model. However, the hostility factor may be improved if 2 items pertaining to suspicion are removed from the scale. These items had relatively low loadings on that factor and decreased the hostility scale's internal reliability. PMID- 7487859 TI - Body size estimation in eating disorder patients: testing the video distortion method on a life-size screen. AB - The video distortion method on a life-size screen is a new method for studying eating disordered patients' estimations of their own body width. Testing this method in female patients (53 with anorexia nervosa, 38 with bulimia nervosa) and 36 normal females, yielded high to very high reliability scores (consistency and test-retest stability). A careful analysis of the 'just noticeable difference' showed that a 5% deviation from the undistorted image should be considered as within the range of accuracy. Taking this into account, anorectic patients appear to score below, above, as well as within the normal range. Bulimia nervosa patients are either accurate in their body estimations or overestimate their body width. All control Ss are correct in their estimations. With this method no significant differences were found between the anorectic patients and the controls. Bulimics differ significantly from anorectics and controls on their optative responses (i.e. what they wanted to look like) and their affective responses (i.e. what they felt they looked like). PMID- 7487861 TI - A system of IAC neural networks as the basis for self-organization in a sociological dynamical system simulation. AB - This sociological simulation uses the ideas of semiotics and symbolic interactionism to demonstrate how an appropriately developed associative memory in the minds of individuals on the microlevel can self-organize into macrolevel dissipative structures of societies such as racial cultural/economic classes, status symbols and fads. The associative memory used is based on an extension of the IAC neural network (the Interactive Activation and Competition network). Several IAC networks act together to form a society by virtue of their human-like properties of intuition and creativity. These properties give them the ability to create and understand signs, which lead to the macrolevel structures of society. This system is implemented in hierarchical object oriented container classes which facilitate change in deep structure. Graphs of general trends and an historical account of a simulation run of this dynamical system are presented. PMID- 7487862 TI - A glimpse of an IPA as a living system. AB - Independent practice association (IPA) model health maintenance organizations (HMO's) represent one of the fastest growing segments of managed care in the United States. IPA's have unique organizational characteristics which thwart efforts to function as a unified whole, particularly where quality of care programs are concerned. Living systems theory was employed as a method to identify subsystems and critical processes within an IPA model HMO in rural California. The application of living systems theory in this way provided insights into the quality of care program within the IPA, which could eventually lead to a redesign of critical processes within the entire system. PMID- 7487863 TI - Computer modeling of adaptive depression. AB - Mild, delimited, and adaptive depression may be a specific example of a more general class of mechanism by which intelligent systems--individual, social, and artificial--adapt to dynamic, uncertain, and dangerous environments. Computer modeling, based on connectionist and artificial intelligence planning and learning programming techniques, supports this hypothesis by generating both adaptive behavior and analogs for 10 phenomena associated with depression: global, stable, and internal failure explantation, a cognitive loop of failure rumination, decreased motivation, self-esteem, and self-efficacy, and increased realism, negative generalization, and cognitive change. The idea of adaptive depression can be applied to more than one level of living systems. A better understanding of normal and adaptive depression may lead to a better understanding of clinical depression. PMID- 7487864 TI - Social skills training for depressed, visually impaired older adults. A treatment manual. AB - Late-onset visual impairment due to cataracts, glaucoma, macular degeneration, or diabetic retinopathy afflicts approximately 10% of people older than 65, and often results in depression and social dysfunction. Whereas the majority of sighted older adults are active participants in their community, individuals suffering from progressively worsening vision experience heightened levels of isolation and reduced social support, and participate in fewer reinforcing recreational activities. This article describes our social skills training package for the treatment of depressed, visually impaired, older adults. Our treatment focuses on increasing the frequency and level of assertiveness with which visually impaired older adults interact with each other. In addition to specified treatment methods, our empirically derived program employs standardized assessment measures to evaluate therapeutic progress. Sessions are implemented to reinforce efforts to apply social skills in the environment and reestablish skills that have begun to fade. PMID- 7487865 TI - Adults' responses to self-injurious behavior. An experimental analysis using a computer-simulation paradigm. AB - The behavior of staff who care for people with mental retardation has been identified as a significant factor in the development and maintenance of challenging behaviors. In a recent analysis, Hastings and Remington (1994a) suggested that both environmental contingencies and rules from other people may affect staff actions. The present study tested this analysis by asking participants to respond to a computer simulation of a work situation involving the care of two individuals who engaged in self-injurious behavior. Fifty participants "interacted" with an attention-seeker and a social-avoider on a simulated teaching task. Results showed that rules were the main factor governing performance. The aversive nature of the contingencies between the self-injury and participants' "attending" behavior also appeared to be influential. The implications of these results for work with care staff, the analysis of challenging behaviors, and experimental research on rule-governed behavior are discussed. PMID- 7487866 TI - The effect of a self-management training package on the transfer of aggression control procedures in the absence of supervision. AB - An aggression replacement and self-management training package reduced the frequency of aggressive behavior among four junior high adolescents identified as seriously emotionally disturbed (SED). During baseline sessions, the students were covertly filmed as they stood unsupervised in front of the school cafeteria. The four subjects engaged in aggressive behavior during 50% of the filmed intervals. These episodes involved provocation by other students, self-initiated provocation, or continuing interaction between students once an aggressive episode had begun. Treatment procedures included instruction, modeling, and role playing of aggression replacement skills. Self-management training included self assessment, self-recording, and self-reinforcement. Following an 8-week period, subjects demonstrated substantial improvement in prosocial skills without supervision. During reversal-to-baseline conditions, aggressive behavior increased; however, reinstating treatment conditions brought a return to prosocial behavior. Outcomes suggest that aggressive replacement skills may transfer and sustain more adequately using self-management. PMID- 7487867 TI - Patient-therapist interaction in the behavioral treatment of panic disorder with agoraphobia. AB - Although effective behavioral techniques have been developed, what aspects of the patient-therapist interaction affect treatment outcome remain largely unknown. This study hypothesized that the interaction between patient and therapist develops over several phases. Further, the association between behavior modes and treatment outcome was expected to alter as that interaction developed. Thirty patients diagnosed with panic disorder with agoraphobia were treated with a standardized behavioral treatment program of 12 sessions. The interpersonal verbal therapist and patient behavior modes were studied at Sessions 1, 3, and 10, using an observational instrument. It was found that behavior modes change over the course of treatment, in line with predictions derived from social psychological models. The hypothesis that establishing a therapeutic relationship requires an empathic and nondirective stance by the therapist in Session 1 was partly confirmed. PMID- 7487869 TI - On the substrate specificity of alpha-crystallin as a molecular chaperone. AB - alpha-Crystallin, the major protein of the ocular lens, acts as a molecular chaperone by preventing the thermal aggregation of proteins. However, in contrast with many other heat shock proteins, alpha-crystallin fails to protect proteins from aggregation during refolding reactions. Our results indicate that alpha crystallin has substrate specificity different from other chaperones and recognizes specific non-native intermediates formed on the denaturation pathway only, with no affinity for intermediates formed on the refolding pathway. PMID- 7487870 TI - Kukoamine A and other hydrophobic acylpolyamines: potent and selective inhibitors of Crithidia fasciculata trypanothione reductase. AB - The enzyme trypanothione reductase (TR), together with its substrate, the glutathione-spermidine conjugate trypanothione, plays an essential role in protecting parasitic trypanosomatids against oxidative stress and is a target for drug design. Here we show that a naturally occurring spermine derivative, the antihypertensive agent kukoamine A [N1N12-bis(dihydrocaffeoyl)-spermine] inhibits TR as a mixed inhibitor (Ki = 1.8 microM, Kii = 13 microM). Kukoamine shows no significant inhibition of human glutathione reductase (Ki > 10 mM) and thus provides a novel selective drug lead. The corresponding N1N8 bis(dihydrocaffeoyl)spermidine derivative was synthesized and acted as a purely competitive inhibitor with Ki = 7.5 microM. A series of mono- and di-acylated spermines and spermidines were synthesized to gain an insight into the effect of polyamine chain length, the nature and position of the acyl substituent and the importance of conformational mobility. These compounds inhibited TR with Ki values ranging from 11 to 607 microM. PMID- 7487871 TI - Reaction mechanism of chitosanase from Streptomyces sp. N174. AB - Chitosanase was produced by the strain of Streptomyces lividans TK24 bearing the csn gene from Streptomyces sp. N174, and purified by S-Sepharose and Bio-Gel A column chromatography. Partially (25-35%) N-acetylated chitosan was digested by the purified chitosanase, and structures of the products were analysed by NMR spectroscopy. The chitosanase produced heterooligosaccharides consisting of D GlcN and GlcNAc in addition to glucosamine oligosaccharides [(GlcN)n, n = 1, 2 and 3]. The reducing- and non-reducing-end residues of the heterooligosaccharide products were GlcNAc and GlcN respectively, indicating that the chitosanase can split the GlcNAc-GlcN linkage in addition to that of GlcN-GlcN. Time-dependent 1H NMR spectra showing hydrolysis of (GlcN)6 by the chitosanase were obtained in order to determine the anomeric form of the reaction products. The chitosanase was found to produce only the alpha-form; therefore it is an inverting enzyme. Separation and quantification of (GlcN)n was achieved by HPLC, and the time course of the reaction catalysed by the chitosanase was studied using (GlcN)n (n = 4, 5 and 6) as the substrate. The chitosanase hydrolysed (GlcN)6 in an endo splitting manner producing (GlcN)2, (GlcN)3 and (GlcN)4, and did not catalyse transglycosylation. Product distribution was (GlcN)3 >> (GlcN)2 > (GlcN)4. Cleavage to (GlcN)3 + (GlcN)3 predominated over that to (GlcN)2 + (GlcN)4. Time courses showed a decrease in rate of substrate degradation from (GlcN)6 to (GlcN)5 to (GlcN)4. It is most likely that the substrate-binding cleft of the chitosanase can accommodate at least six GlcN residues, and that the cleavage point is located at the midpoint of the binding cleft. PMID- 7487872 TI - Rapid desensitization of the thyrotropin-releasing hormone receptor expressed in single human embryonal kidney 293 cells. AB - This study uses fluorescence microscopy combined with dynamic video imaging to examine the events associated with the rapid desensitization of the thyrotropin releasing hormone receptor (TRH-R). In single non-pituitary human embryonic kidney 293 (HEK-293) cells, expressing either the rat or human TRH-Rs, TRH produced a rapid dose-dependent monophasic rise in [Ca2+]i. This Ca2+ transient was completely abolished by pretreatment of cells with the intracellular Ca2+ antagonists thapsigargin or cyclopiazonic acid, but not EGTA, the voltage operated Ca2+ channel (VOCC) antagonist nifedipine or the second-messenger operated Ca2+ channel antagonist SK&F 96365. These results suggest that TRH causes the mobilization of Ca2+ from thapsigargin/cyclopiazonic acid-sensitive intracellular Ca2+ stores but not the influx of extracellular Ca2+. HEK-293 cells also failed to respond to KCl or the slow Ca(2+)-channel activator BAY K 8644, suggesting that they lack L-type VOCCs. Rat and human TRH-Rs are highly conserved except at the C-terminus where the sequence differs. The C-terminus is believed to be important in receptor desensitization. Despite differences in this region, rat and human TRH-Rs expressed in HEK-293 cells underwent rapid (within 1 min) desensitization. This desensitization was dose-dependent and did not involve receptor loss. Similarly the bradykinin receptor endogenous to HEK-293 cells also displays a rapid desensitization. We conclude that in TRH-R-expressing non pituitary HEK-293 cells, TRH mobilizes intracellular Ca2+ resulting in a monophasic Ca2+ transient. The rat and human TRH-Rs as well as the endogenous bradykinin receptor also displayed rapid receptor desensitization. PMID- 7487868 TI - Structure and function of the aromatic amino acid hydroxylases. PMID- 7487873 TI - Transforming growth factor-beta 1 increases internalization of basic fibroblast growth factor by smooth muscle cells: implication of cell-surface heparan sulphate proteoglycan endocytosis. AB - Basic fibroblast growth factor (bFGF) was internalized by smooth muscle cells (SMC) from pig aorta. Correlation between heparin inhibition of binding and late internalization (8 h) implicated low-affinity sites in bFGF internalization. Transforming growth factor-beta 1 (TGF-beta 1) induced a 38% increase in bFGF internalized between 4 and 8 h. While bFGF and/or TGF-beta 1 enhanced cell surface proteoglycan synthesis, 35S-labelled proteoglycans of the extracellular matrix (ECM) were not affected. This might be explained by the different turnover rates displayed by the two populations of proteoglycans. Although bFGF and/or TGF beta 1 induced a similar stimulation in cell-surface chondroitin sulphate/dermatan sulphate and heparan sulphate (HS) proteoglycan synthesis, only the turnover of HS proteoglycans was increased. Twice as much HS proteoglycan was internalized in the presence of TGF-beta 1 or bFGF. Furthermore, TGF-beta 1 induced a 43 +/- 12% increase in HS proteoglycan internalized in the presence of bFGF with a parallel 38% increase in bFGF internalization. Overall, the results indicated that bFGF bound to two HS proteoglycan populations. bFGF storage (70% of bFGF bound to SMC) was not affected by TGF-beta 1 under our conditions and involved ECM proteoglycans characterized by a low turnover. bFGF internalization up-regulated by TGF-beta 1 involved cell-surface HS proteoglycan characterized by a high turnover. PMID- 7487876 TI - Protochlorophyllide reductase in photosynthetic prokaryotes and its role in chlorophyll synthesis. AB - 1. DNA sequences hybridizing with the wheat protochlorophyllide reductase gene have been detected during genomic Southern blots of various cyanobacterial DNA samples. No such hybridization was observed with DNA from photosynthetic bacteria. 2. A fragment amplified from Phormidium laminosum DNA has been characterized and shown to be 73% similar to the corresponding wheat sequence. At the protein level the similarity is 91%. When used as a probe for Southern blotting the Phormidium DNA fragment confirmed the authenticity of some of the original signals obtained with the wheat probe. 3. Peptides of molecular mass 36, 30 and 60 kDa are immunodetected by a wheat reductase antibody during Western blotting of Phormidium preparations. These are purported to correspond to the cyanobacterial mature reductase protein, a stable proteolytic fragment and soluble dimeric forms of the latter respectively. 4. Adaptation of Phormidium to growth in red light (delta > 670 nm) or darkness led to no significant changes in the total level of immunodetected peptides or protochlorophyllide within the cells. 5. The specific activity of the reductase in Phormidium membranes has been tentatively estimated as 0.5 unit/mg of protein, a value comparable with that found in preparations from mature chloroplasts of higher plants. PMID- 7487875 TI - Purification and characterization of a uterine retinol-binding protein in the bitch. AB - A major canine endometrial secreted protein (cP6, 23,000-M(r)) was purified by ion-exchange and gel-filtration chromatography and characterized by two dimensional gel electrophoresis. Anti-[human retinol-binding protein (hRBP)] serum identified cP6 on immunoblot analysis and immunoprecipitated cP6 from culture medium. This major protein was also shown to bind [3H]retinol. N-terminal and internal amino acid sequences were determined and compared with previously identified protein, RNA, or DNA sequences. N-terminal analysis revealed that cP6 had high identity and similarity to serum retinol-binding proteins (RBPs), while internal sequence analysis showed a strong similarity to rat androgen-dependent epididymal protein and beta-lactoglobulins. Amino acid analysis, however, showed significant differences between these proteins and cP6 in both total amino acid content and certain selected amino acids. Immunohistochemical analysis showed staining for RBP only in the uterine luminal epithelium. These studies suggest that bitch endometrium secretes a family of proteins (cP6), some of which bind [3H]retinol, are immunologically related to the RBP family, and have N-terminal and internal sequences with a high similarity to RBP, beta-lactoglobulins and other members of the lipocalin family. This family of proteins may be important in early development for supplying retinol or derivatives to the developing embryo. PMID- 7487874 TI - Stimulatory antibody-induced activation and selective translocation of protein kinase C isoenzymes in human platelets. AB - A novel stimulatory monoclonal antibody (Mab) termed Mab.F11 induces granular secretion and subsequent aggregation of human platelets. Mab.F11 recognizes a unique 32 and 35 kDa protein duplex on the platelet membrane surface, called the F11 receptor; binding of Mab.F11 to its receptor results in increased intracellular phosphorylation of P47, the known protein kinase C (PKC) substrate pleckstrin. In order to determine whether the mechanism of action of Mab.F11 involves direct activation of PKC, two types of functional assays for measuring PKC activity were performed. Measurement of PKC activity in digitonin permeabilized platelets revealed that Mab.F11 produced a rapid, 2-3 fold increase in the control value in the phosphorylation of the PKC peptide substrate, PKC(19 31) Ser25. The increase in PKC activity induced by Mab.F11 was found to be associated with the platelet membrane; a 1.6-fold control value increase in membrane PKC activity occurred rapidly, within 10 s of the addition of Mab.F11. The translocation from the cytoplasm to the membrane induced by Mab.F11 in PKC isoenzymes alpha and zeta was reversible, whereas translocation of the PKC isoenzymes delta, beta, eta' and theta was irreversible, with PKC levels remaining elevated in the membrane for at least 15 min. Taken together, our results demonstrate that in the initial stages of platelet activation by this stimulatory antibody, the enhanced membrane PKC activity reflects the presence of all six isoenzymes. At later stages, PKC activity is reflective of four isoenzymes. These results demonstrate that separate groups of PKC isoenzymes must be involved in different aspects of platelet activation. The long lag period and prolonged activation time of platelets by Mab.F11 renders this agonist most suitable for identifying the isoenzymes and their specific endogenous protein substrates involved in platelet secretion and aggregation induced by platelet membrane protein antibodies. PMID- 7487877 TI - Tissue-specific regulation of selenoenzyme gene expression during selenium deficiency in rats. AB - Regulation of synthesis of the selenoenzymes cytosolic glutathione peroxidase (GSH-Px), phospholipid hydroperoxide glutathione peroxidase (PHGSH-Px) and type-1 iodothyronine 5'-deiodinase (5'IDI) was investigated in liver, thyroid and heart of rats fed on diets containing 0.405, 0.104 (Se-adequate), 0.052, 0.024 or 0.003 mg of Se/kg. Severe Se deficiency (0.003 mg of Se/kg) caused almost total loss of GSH-Px activity and mRNA in liver and heart. 5'IDI activity decreased by 95% in liver and its mRNA by 50%; in the thyroid, activity increased by 15% and mRNA by 95%. PHGSH-Px activity was reduced by 75% in the liver and 60% in the heart but mRNA levels were unchanged; in the thyroid, PHGSH-Px activity was unaffected by Se depletion but its mRNA increased by 52%. Thus there is differential regulation of the three mRNAs and subsequent protein synthesis within and between organs, suggesting both that mechanisms exist to channel Se for synthesis of a particular enzyme and that there is tissue-specific regulation of selenoenzyme mRNAs. During Se depletion, the levels of selenoenzyme mRNA did not necessarily parallel the changes in enzyme activity, suggesting a distinct mechanism for regulating mRNA levels. Nuclear run-off assays with isolated liver nuclei showed severe Se deficiency to have no effect on transcription of the three genes, suggesting that there is post-transcriptional control of the three selenoenzymes, probably involving regulation of mRNA stability. PMID- 7487878 TI - Synthesis and mass-spectrometric characterization of human serum albumins modified by covalent binding of two non-steroidal anti-inflammatory drugs: tolmetin and zomepirac. AB - Human serum albumins modified by covalently bound tolmetin or zomepirac were synthesized as models for similar products formed in vivo from acyl glucuronides. Activated esters of both drugs were prepared with 1-ethyl-3-(3 dimethylaminopropyl)-carbodi-imide, and then allowed to react with human serum albumin. Tryptic digests of both protein products were analysed by HPLC to identify peptides containing covalently bound drugs, and binding sites on albumin were identified by high-performance tandem MS. Three binding sites were common to both products, i.e. lysine-195, -199 and -351. Three further modified residues were identified for the tolmetin-albumin product, i.e. aspartic acid 1, and lysine-524 and -536. PMID- 7487880 TI - Co-localization and functional coupling of creatine kinase B and gastric H+/K(+) ATPase on the apical membrane and the tubulovesicular system of parietal cells. AB - Immunogold labelling of creatine kinase B (BB-CK) and gastric H+/K(+)-ATPase in the parietal cells of the stomach revealed colocalization of these two enzymes on the apical membrane and the membranes of the tubulovesicular system. Upon fractionation of hog parietal cells, a specific fraction of the BB-CK proteins remained associated with the purified vesicles, in which gastric H+/K(+)-ATPase is highly enriched. The BB-CK present in this highly purified preparation was able to support pronounced H+/K(+)-ATPase activity in K(+)-loaded vesicles in the presence of phosphocreatine and ADP, although only low levels of ATP were measured. In contrast, when pyruvate kinase, phosphoenolpyruvate and ADP were used as an ATP-generating system to sustain similar levels of H+/K(+)-ATPase activity, ATP levels were more than 10-fold higher. Changing the experimental conditions such that ATP levels were the same for both systems resulted in significantly elevated H+/K(+)-ATPase activities in the BB-CK/phosphocreatine system in comparison with the pyruvate kinase/phosphoenolpyruvate system. These results indicate that gastric H+/K(+)-ATPase has preferential access to ATP generated by creatine kinase co-localized on the membranes of the vesicles. PMID- 7487879 TI - Molecular cloning of a novel widely expressed human 80 kDa 17 beta-hydroxysteroid dehydrogenase IV. AB - Reactions of oestrogens and androgens at position C-17 are catalysed by 17 beta hydroxysteroid dehydrogenases (17 beta-HSDs). Cloning of the cDNA of a novel human 17 beta-HSD IV and expression of its mRNA are described. A probe derived from the recently discovered porcine 17 beta-oestradiol dehydrogenase (17 beta EDH) was used to isolate a 2.6 kb human cDNA encoding a continuous protein of 736 amino acids of high (84%) similarity to the porcine 17 beta-EDH. The calculated molecular mass of the human enzyme is 79,595 Da. Other sequence similarities shared by the two enzymes are: an N-terminal sequence which is similar to that of members of the short-chain alcohol dehydrogenase family; amino acids 343-607 which are similar to the C-terminal domains of a trifunctional Candida tropicalis enzyme and the FOX2 gene product of Saccharomyces cerevisiae; amino acids 596-736 which are similar to human sterol carrier protein 2. The previously cloned human 17 beta-HSD I, II and III are less than 25% identical with 17 beta-HSD IV. mRNA for HSD IV is a single species of 3.0 kb, present in many tissues with highest concentrations in liver, heart, prostate and testes. When over-expressed in mammalian cells, the human 17 beta-HSD IV enzyme displays a specific unidirectional oxidative 17 beta-HSD activity. PMID- 7487881 TI - Isothiocyanates as substrates for human glutathione transferases: structure activity studies. AB - The catalytic properties of four human glutathione transferases (GSTs), A1-1, M1 1, M4-4 and P1-1, were examined with 14 isothiocyanate (R-NCS) substrates. The compounds include aliphatic and aromatic homologues, some of which are natural constituents of human food, namely sulphoraphane [1-isothiocyanato-4 (methylsulphinyl)butane], erucin [1-isothiocyanato-4-(methylthio)butane], erysolin [1-isothiocyanato-4-(methylsulphonyl)butane], benzyl-NCS, phenethyl-NCS and allyl-NCS. All isothiocyanates investigated were substrates for the four GSTs. The enzymes promote addition of the thiol group of GSH to the electrophilic central carbon of the isothiocyanate group to form dithiocarbamates [R-NH-C(=S) SG] which have high UV absorption at 274 nm. Molar absorption coefficients and non-enzymic rate constants as well as standardized enzyme assay conditions for all compounds were established. Of the four isoenzymes investigated, GSTs M1-1 and P1-1 were generally the most efficient catalysts, whereas GST M4-4 was the least efficient. Isothiocyanates are among the GST substrates that are most rapidly conjugated. On the basis of rate-enhancement data and binding energies, the isothiocyanates were compared with 4-hydroxyalkenals, another class of natural GST substrates previously subjected to systematic kinetic analysis. The incremental transition-state stabilization attributable to an increased number of methylene groups in homologous alkyl isothiocyanates is similar to that previously noted for homologous 4-hydroxyalkenals. PMID- 7487882 TI - Identification of a heparin-binding protein using monoclonal antibodies that block heparin binding to porcine aortic endothelial cells. AB - The binding of heparin or heparan sulphate to a variety of cell types results in specific changes in cell function. Endothelial cells treated with heparin alter their synthesis of heparan sulphate proteoglycans and extracellular matrix proteins. In order to identify a putative endothelial cell heparin receptor that could be involved in heparin signalling, anti-(endothelial cell) monoclonal antibodies that significantly inhibit heparin binding to endothelial cells were prepared. Four of these antibodies were employed in affinity-chromatographic isolation of a heparin-binding protein from detergent-solubilized endothelial cells. The heparin-binding protein isolated from porcine aortic endothelial cells using four different monoclonal antibodies has an M(r) of 45,000 assessed by SDS/PAGE. The 45,000-M(r) heparin-binding polypeptide is isolated as a multimer. The antibody-isolated protein binds to heparin-affinity columns as does the pure 45,000-M(r) polypeptide, consistent with its identification as a putative endothelial heparin receptor. PMID- 7487883 TI - Syk interacts with tyrosine-phosphorylated proteins in human platelets activated by collagen and cross-linking of the Fc gamma-IIA receptor. AB - Activation of human platelets by cross-linking of the platelet low-affinity IgG receptor, the Fc gamma receptor IIA (Fc gamma-RIIA), or by collagen is associated with rapid phosphorylation on tyrosine of the non-receptor tyrosine kinase syk. Phosphorylation is still observed, albeit sometimes reduced, in the presence of a combination of a protein kinase C inhibitor, Ro 31-8220, and the intracellular calcium chelator, BAPTA-AM, demonstrating independence from phosphoinositide specific phospholipase C (PLC) activity. In contrast, the combination of Ro 31 8220 and BAPTA-AM completely inhibits phosphorylation of syk in thrombin stimulated platelets. Phosphorylation of syk increases its autophosphorylation activity measured in a kinase assay performed on syk immunoprecipitates. Fc gamma RIIA also undergoes phosphorylation in syk immunoprecipitates from platelets activated by cross-linking of Fc gamma-RIIA but not by collagen, suggesting that it associates with the kinase. Consistent with this, tyrosine-phosphorylated Fc gamma-RIIA is precipitated by a glutathione S-transferase (GST) fusion protein containing the tandem src homology (SH2) domains of syk from Fc gamma-RIIA- but not collagen-activated cells. Two uncharacterized tyrosine-phosphorylated proteins of 40 and 65 kDa are uniquely precipitated by a GST fusion protein containing the tandem syk-SH2 domains in collagen-stimulated platelets. A peptide based on the antigen recognition activation motif (ARAM) of Fc gamma-RIIA, and phosphorylated on the two tyrosine residues found within this region, selectively binds syk from lysates of resting platelets; this interaction is not seen with a non-phosphorylated peptide. Kinase assays on Fc gamma-RIIA immunoprecipitates reveal the constitutive association of an unidentified kinase activity in resting cells which phosphorylates a 67 kDa protein. Syk is not detected in Fc gamma-RIIA immunoprecipitates from resting cells but associates with the receptor following activation and, together with Fc gamma-RIIA, is phosphorylated in the kinase assay in vitro. These results demonstrate that syk is activated by Fc gamma-RIIA cross-linking and collagen, independent of PLC, suggesting that it may have an important role in the early events associated with platelet activation. The association of syk with Fc gamma-RIIA appears to be mediated through the tandem SH2 domains in syk and the ARAM motif of Fc gamma-RIIA. A similar interaction may underlie the response to collagen, suggesting that its signalling receptor contains an ARAM motif. PMID- 7487884 TI - Reconstitution and characterization of ATP-dependent bile acid transport in human and rat placenta. AB - Bile acid (BA) transport across the human microvillus maternal-facing trophoblast plasma membrane (mTPM) has been recently reported to be stimulated by the presence of ATP [Marin, Bravo, El-Mir and Serrano (1993) J. Hepatol. 18, S41]. Reconstitution of BA transport activity in proteoliposomes from human mTPM is reported in this paper. Typical characteristics of BA transport in native mTPM vesicles, including a requirement for ATP hydrolysis and inhibition by other BA species, were preserved in proteoliposome preparations. BA transport into 20- and 14-day-gestation rat mTPM vesicles was also stimulated by the presence of ATP as noted in human mTPM and in the rat liver canalicular membrane. Besides this functional similarity, these ATP-dependent carriers may share structural characteristics, as demonstrated by studies using an antibody (100 Ab) raised against the 100 kDa BA carrier of the canalicular membrane from rat liver which recognized proteins in both human and rat brush-border trophoblast membranes. PMID- 7487885 TI - The roles of the conserved pyrimidine bases in hammerhead ribozyme catalysis: evidence for a magnesium ion-binding site. AB - We report details of the synthesis and characterization of oligoribonucleotides containing 4-thiouridine or 2-pyrimidinone ribonucleoside (4HC). We have used these probes to examine the roles of the conserved pyrimidines in the central core of the hammerhead ribozyme. The effects on catalysis of singly-substituted hammerhead ribozyme and substrate strands were quantified in multiple-turnover reactions. Various effects were observed on kcat. and Km, with up to a 7-fold decrease and a 3-fold increase respectively. For substitutions with 4HC at positions 3 or 17, catalytic activity in single turnover reactions can be increased up to 8-fold equivalent to 40% of wild-type activity, by increasing the concentration of the Mg2+ cofactor, implying that these substitutions had a deleterious effect on Mg2+ binding. Calculations of the change in the apparent free energy of binding for variants at positions 3, 4 or 17 are each consistent with deletion of a single hydrogen-bond to an uncharged group in the ribozyme. The cytidine 5' to the scissile phosphate had not previously been thought to play a direct role in catalysis, however, removal of the exocyclic amino group decreased kcat. 4-fold. Recently, the crystal structures of a hammerhead ribozyme bound to either a non-cleavable 2'-deoxy substrate strand or a ribo-substrate strand have been reported. The kinetic properties of the variants described here are consistent with several key interactions seen in the crystals, in particular they provide experimental support for the assignment of the proposed catalytically active magnesium ion-binding site. PMID- 7487887 TI - Mutation of an EF-hand Ca(2+)-binding motif in phospholipase C of Dictyostelium discoideum: inhibition of activity but no effect on Ca(2+)-dependence. AB - Phosphoinositide-specific phospholipase C (PLC) is dependent on Ca2+ ions for substrate hydrolysis. The role of an EF-hand Ca(2+)-binding motif in Ca(2+) dependent PLC activity was investigated by site-directed mutagenesis of the Dictyostelium discoideum PLC enzyme. Amino acid residues with oxygen-containing side chains at co-ordinates x, y, z, -x and -z of the putative Ca(2+)-binding loop sequence were replaced by isoleucine (x), valine (y) or alanine (z, -x and z). The mutated proteins were expressed in a Dictyostelium cell line with a disrupted plc gene displaying no endogenous PLC activity, and PLC activity was measured in cell lysates at different Ca2+ concentrations. Replacement of aspartate at position x, which is considered to play an essential role in Ca2+ binding, had little effect on Ca2+ affinity and maximal enzyme activity. A mutant with substitutions at both aspartate residues in position x and y also showed no decrease in Ca2+ affinity, whereas the maximal PLC activity was reduced by 60%. Introduction of additional mutations in the EF-hand revealed that the Ca2+ concentration giving half-maximal activity was unaltered, but PLC activity levels at saturating Ca2+ concentrations were markedly decreased. The results demonstrate that, although the EF-hand domain is required for enzyme activity, it is not the site that regulates the Ca(2+)-dependence of the PLC reaction. PMID- 7487886 TI - The glycoinositol-phospholipids of Phytomonas. AB - The Phytomonas spp. are trypanosomatid parasites of plants. A polar glycolipid fraction of a Phytomonas sp., isolated from the plant Euphorbia characias and grown in culture, was fractionated into four major glycolipid species (Phy 1-4). The glycolipids were analysed by chemical and enzymic modifications, composition and methylation analyses, electrospray mass spectrometry and microsequencing after HNO2 deamination and NaB3H4 reduction. The water-soluble headgroup of the Phy2 glycolipid was also analysed by 1H NMR. All four glycolipids were shown to be glycoinositol-phospholipids (GIPLs) with phosphatidylinositol (PI) moieties containing the fully saturated alkylacylglycerol lipids 1-O-hexadecyl-2-O palmitoylglycerol and 1-O-hexadecyl-2-O-stearoylglycerol. The structures of the Phy 1-4 GIPLs are: Man alpha 1-2Man alpha 1-6Man alpha 1-4GlcN alpha 1-6PI, Glc alpha 1-2(NH2-CH2CH2-HPO4-)Man alpha 1-2Man alpha 1-6Man alpha 1-4GlcN alpha 1 6PI, [formula: see text] Glc alpha 1-2(NH2CH2CH2-HPO4-)Man alpha 1-2Man alpha 1 6Man alpha 1-4(NH2-CH2CH2-HPO4-)GlcN alpha 1-6PI [formula: see text] and Glc alpha 1-2Glc alpha 1-2(NH2CH2-CH2-HPO4-)Man alpha 1-2Man alpha 1-6Man alpha 1 4(NH2CH2CH2-HPO4-)-GlcN alpha 1-6PI. [formula: see text] The Phytomonas GIPLs represent a novel series of structures. This is the first description of the chemical structure of cell-surface molecules of this plant pathogen. The Phytomonas GIPLs are compared with those of other trypanosomatid parasites and are discussed with respect to trypanosomatid phylogenetic relationships. PMID- 7487888 TI - The mRNAs for the three chains of human collagen type XI are widely distributed but not necessarily co-expressed: implications for homotrimeric, heterotrimeric and heterotypic collagen molecules. AB - In cartilage collagen type XI exists as heterotrimeric molecules composed of alpha 1(XI), alpha 2(XI) and alpha 3(XI) subunits. Messenger RNAs for some of the alpha chains of collagen type XI have also been found in non-chondrogenic tissues but the chain composition of the molecule in these sites is not known. Some non chondrogenic tissues also contain heterotrimers containing collagen alpha 2(V) and alpha 1(XI) chains. We have explored the possibility that collagen type XI could exist in differing trimeric forms in non-chondrogenic tissues and aimed to predict the subunit composition of this collagen in those tissues. The distribution and relative levels of expression of collagen alpha 1(XI), alpha 2(XI) and alpha 3(XI)/alpha 1(II) mRNAs in different human fetal tissues were studied. Expression of mRNAs for all three genes of collagen type XI is not restricted to cartilage but is widespread. However, in some non-chondrogenic tissues, the mRNAs for all three alpha chains of collagen type XI were not co expressed, but collagen alpha 1(XI) and alpha 2(XI) mRNAs were found either singly or without collagen alpha 3(XI) transcripts. Collagen type XI may therefore exist as homotrimers and/or heterotrimers composed of two collagen alpha(XI) chains in some tissues. The distribution of mRNAs for collagen alpha 2(V) and alpha 1(I) were also studied. Co-expression of collagen type XI, alpha 2(V) and alpha 1(I) mRNAs was found for many tissues. These findings have implications for the possibility of additional chain associations for collagen types XI and V in cross-type heterotrimers within heterotypic fibrils. PMID- 7487889 TI - Carrier-mediated fructose uptake significantly contributes to carbohydrate metabolism in human skeletal muscle. AB - To determine whether fructose can be utilized as a metabolic substrate for skeletal muscle in man, we investigated its incorporation into glycogen, its oxidation and lactate production in isolated human skeletal muscle. Rates of fructose oxidation and incorporation into glycogen increased in the presence of increasing fructose concentrations (0.1-1.0 mM). Lactate production increased 3 fold when extracellular fructose was increased from 0.1 to 0.5 mM. Cytochalasin B, a competitive inhibitor of hexose transport mediated by the GLUT1 and GLUT4 facilitative glucose transporters, completely inhibited insulin-stimulated glucose incorporation into glycogen and glucose oxidation (P < 0.01), but did not alter fructose incorporation into glycogen or fructose oxidation. Insulin (1000 mu-units/ml) increased glucose incorporation into glycogen 2.7-fold and glucose oxidation 2.3-fold, whereas no effect on fructose incorporation into glycogen or fructose oxidation was noted. A physiological concentration of glucose (5 mM) decreased the rate of 0.5 mM fructose incorporation into glycogen by 60% (P < 0.001), whereas fructose oxidation was not altered in the presence of 5 mM glucose. Irrespective of fructose concentration, the majority of fructose taken up underwent non-oxidative metabolism. Lactate production accounted for approx. 80% of the fructose metabolism in the basal state and approx. 70% in the insulin (1000 mu-units/ml)-stimulated state. In the presence of 5 mM glucose, physiological concentrations of fructose could account for approximately 10-30% of hexose (glucose + fructose) incorporation into glycogen under non-insulin stimulated conditions. In conclusion, fructose appears to be transported into human skeletal muscle via a carrier-mediated system that does not involve GLUT4 or GLUT1. Furthermore, under physiological conditions, fructose can significantly contribute to carbohydrate metabolism in human skeletal muscle. PMID- 7487890 TI - Impairment of the liver insulin receptor autoactivation cascade at full-term pregnancy in the rat. AB - Partially purified liver insulin receptors from full-term pregnant rats show decreased autophosphorylation rates if compared with receptors from virgins. We studied the molecular mechanism of this phenomenon, looking at possible structural and functional changes of several domains. The ATP-binding domain seems to be unaltered in receptors from pregnant rats since Km for ATP was similar to that observed in virgins. In contrast, the Vmax. is decreased some 45%, suggesting changes in the kinase domain. Truncation of a fragment of 10 kDa from the C-terminal tail does not normalize the kinase activity in receptors from pregnant rats, suggesting that this domain is not involved in the inhibitory regulation. Treatment with alkaline phosphatase increases the [32P]Pi incorporation into receptors from pregnant rats; however, the autophosphorylation remains lower than that observed in virgin rats. Tryptic phosphopeptide maps of phosphorylated receptors show that the same phosphopeptides are present in receptors from virgin and pregnant rats. However, the progression through the autoactivation cascade in the kinase domain is impaired in receptors from pregnant rats. Differences in the cleavage by trypsin at the two alternative sites in the kinase domain were observed, indicating possible structural changes in receptors from pregnant rats that could be related to the impairment of the autoactivation cascade. Integrity of the alpha- and beta-subunits, as well as differential expression of the two receptor isotypes, were shown to be unaltered. We conclude that (1) the decreased autophosphorylation rate of the liver insulin receptor from pregnant rats is associated with the impairment of its autoactivation cascade, probably as a consequence of the basal Ser/Thr phosphorylation; and (2) the inhibition of the autoactivation cascade does not account for the overall inhibition of autophosphorylation observed in receptors from pregnant rats. PMID- 7487891 TI - Pretranslational regulation of pyruvate dehydrogenase complex subunits in white adipose tissue during the suckling-weaning transition in the rat. AB - Total pyruvate dehydrogenase complex activity is low in white adipose tissue during the suckling period and increases markedly at weaning on to a high carbohydrate diet. This is concomitant with an increase in the E1 alpha, E1 beta and E2 subunit protein concentration and their respective mRNAs, suggesting a pretranslational control of this phenomenon. The most marked change is seen for the E1 alpha subunit (17-fold increase in protein concentration). The changes in pyruvate dehydrogenase complex activity and subunit abundance induced by weaning on to a high-carbohydrate diet are precluded if the animals are weaned on to a high-fat diet, suggesting that the nutritional and/or related hormonal changes rather than a developmental stage are responsible for the observed adipose-tissue pyruvate dehydrogenase complex pattern. PMID- 7487892 TI - An altered T2 beta translocase of the glucose-6-phosphatase system in the membrane of the endoplasmic reticulum from livers of Ehrlich-ascites-tumour bearing mice. AB - The inhibitory interactions of orthophosphate (P1) with the glucose-6-phosphatase system of intact microsomes derived from the livers of normal and Ehrlich-ascites tumour-bearing mice reveal the appearance of a novel form of the T2 beta translocase component of the glucose-6-phosphatase system in tumour-stressed mice. Kinetic studies, with and without 20 mM P1, show a strictly classical competitive inhibition, with a K1,P1 of 4.2 mM, with disrupted microsomes from both control and tumour-bearing mouse liver. Inhibition was also observed with intact microsomes from livers of control mice, and contributions by both competitive and non-competitive components of inhibition were quantified by calculation of Kis,P1 and Kii,P1 values respectively. However, little inhibition was noted with intact microsomes from the livers of tumour-bearing mice. It is concluded that this novel form of T2 beta is less able to transport Pi, from the cytosol to the endoplasmic reticulum lumen, perhaps because of the tumour-related increased Km for Pi transport in this direction. PMID- 7487893 TI - A negative regulatory element in the promoter region of the rat alpha 2A adrenergic receptor gene overlaps an SP1 consensus binding site. AB - Three subtypes of alpha 2-adrenergic receptors (alpha 2A, alpha 2B and alpha 2C) have been described that differ in their primary sequence and tissue-specific expression and are encoded by three distinct genes. Previous work has shown that the human alpha 2A-adrenergic receptor gene promoter consists of a TATA-box (TATAAA), palindromic sequence (CCCACGTGGG) and GC-box (GGGGCGG) motif. Sequence analysis of the putative promoter region of the rat alpha 2A-adrenergic receptor gene showed that these promoter regions are conserved in their sequence and relative location. We analysed the transcriptional activity of these regions using RINm5F, a rat insulinoma cell line that expresses the endogenous alpha 2A adrenergic receptor gene. These results showed that the region from -484 to -92 has a negative effect on transcription, as deletion of this region in alpha 2A adrenergic receptor gene-chloramphenicol acetyltransferase reporter constructs increased reporter gene activity. This region included the GC-box sequence which is a consensus binding site for the nuclear factor SP1, which is a positive activator of transcription. Gel-mobility-shift assays and supershift assays with an antibody that recognizes SP1 showed binding of the SP1 nuclear factor as well as other nuclear factors to this GC-box region. Additional nuclear factors bind to the downstream palindromic region. We suggest that positive- and negative acting nuclear factors contribute to the activity of the alpha 2-adrenergic receptor promoter. PMID- 7487895 TI - Insulin stimulation of intracellular free Ca2+ recovery and Ca(2+)-ATPase gene expression in cultured vascular smooth-muscle cells: role of glucose 6-phosphate. AB - Wa have previously reported that insulin accelerates recovery of intracellular Ca2+ concentrations ([Ca2+]i) from pressor agonist-induced Ca2+ loads and stimulates both plasmalemmal and sarcoplasmic-reticulum Ca(2+)-ATPase gene expression in cultured and freshly isolated vascular smooth-muscle cells (VSMCs), suggesting that insulin attenuation of vascular tone may result from modulation of [Ca2+]i. Accordingly, we have now evaluated the linkage between this insulin regulation of VSMC[Ca2+]i and classical actions of insulin (i.e. glucose transport and metabolism). Cultured VSMCs were incubated in the presence or absence of insulin in a medium containing either pyruvate, glucose, 3-O methylglucose or 2-deoxyglycose. Insulin caused an 87% increase in [Ca2+]i recovery rate after stimulation with arginine-vasopressin (P < 0.01) and caused a marked increase in Ca(2+)-ATPase mRNA and protein levels in the presence of glucose. Comparable increases in both [Ca2+]i recovery and Ca2(+)-ATPase expression were found when glucose was replaced by 2-deoxyglucose. In contrast, no stimulation was found in either the glucose-free or 3-O-methylglucose containing medium. As both glucose analogues are transported, but only 2 deoxyglucose is phosphorylated, this indicates that glucose transport and metabolism to glucose 6-phosphate is essential for insulin regulation of VSMC [Ca2+]i, possibly via a glucose-6-phosphate-dependent carbohydrate-response element in the Ca2(+)-ATPase gene. PMID- 7487894 TI - Structure of the human TIMP-3 gene and its cell cycle-regulated promoter. AB - The gene encoding tissue inhibitor of metalloproteinases-3 (TIMP-3) is regulated during development, mitogenic stimulation and normal cell cycle progression. The TIMP-3 gene is structurally altered or deregulated in certain diseases of the eye and in tumour cells. A detailed knowledge of the TIMP-3 gene and its regulatory elements is therefore of paramount importance to understand its role in development, cell cycle progression and disease. In this study, we present the complete structure of the human TIMP-3 gene. We show that TIMP-3 is a TATA-less gene, which initiates transcription at one major site, is composed of five exons and four introns spanning a region of approximately 30 kb, and gives rise to three distinct mRNAs, presumably due to the usage of alternative polyadenylation signals. Using somatic cell hybrids the TIMP-3 locus was mapped to chromosomal location 22q13.1 We also show that the TIMP-3 5' flanking region is sufficient to confer both high basal level expression in growing cells and cell cycle regulation in serum-stimulated cells. While the first 112 bases of the promoter, which harbour multiple Sp1 sites, were found to suffice for high basal level activity, the adjacent region spanning positions -463 and -112 was found to be a major determinant of serum inducibility. These results provide an important basis for further investigations addressing the role of TIMP-3 in physiological processes and pathological conditions. PMID- 7487896 TI - Identification of a new membrane-bound heparan sulphate proteoglycan. AB - The morphological changes that occur during intestinal development have been extensively described, but the molecular basis of these changes is largely unknown. As a result of our efforts to identify molecules that play a role in intestinal morphogenesis during development, we have previously isolated a cDNA that is developmentally regulated in the intestine. This cDNA, named OCI-5, was recently shown to have 20-25% identity at the protein-sequence level with glypican and cerebroglycan, two heparan sulphate proteoglycans (HSPG) that are attached to the cell membrane by a glycosyl-phosphatidylinositol (GPI) anchor. Here we provide experimental evidence indicating that OCI-5 is also a GPI-linked HSPG. We demonstrate this by showing that OCI-5 can be labelled with radioactive sulphate and can be digested by heparitinase, but not by chondroitinase. We also show that treatment with phosphatidylinositol-specific phospholipase C releases OCI-5 from the cell surface of COS cells transfected with an OCI-5 expression vector. The identification of OCI-5 as a GPI-linked HSPG confirms that this proteoglycan belongs to the same family of HSPGs that include glypican and cerebroglycan. PMID- 7487897 TI - Differential modulation of expression of the two acylphosphatase isoenzymes by thyroid hormone. AB - The modulation of expression of the skeletal muscle and erythrocyte acylphosphatase isoenzymes by thyroid hormone has been investigated. Our results indicate a differential regulation of the two enzymic isoforms by tri iodothyronine (T3) in K562 cells in culture: an increase in the specific mRNA during T3-stimulation is shown only for the skeletal muscle isoenzyme. A fast and transient T3 induction of the accumulation of the specific mRNA can be observed, reaching a maximum 8 h after hormone treatment and then rapidly decreasing almost to the steady-state level after 24 h. A nuclear run-on assay was performed to explore the mechanisms of this regulation. These studies indicate that T3 induction of skeletal muscle acylphosphatase mRNA is due, at least in part, to a fast and transient increase in the rate of gene transcription, within 4 h after hormone administration. A very rapid decrease is then observed within a further 2 h. T3-dependent accumulation of the mRNA for the skeletal muscle acylphosphatase requires ongoing protein synthesis, as confirmed by inhibition with cycloheximide or puromycin. These findings indicate that the transcriptional regulation of the gene may be indirect. PMID- 7487898 TI - 5-hydroxytryptamine stimulates glucose transport in cardiomyocytes via a monoamine oxidase-dependent reaction. AB - This study deals with the effect of 5-hydroxytryptamine (5-HT; serotonin) on glucose transport in isolated rat cardiac myocytes. In these cells, 5-HT (10-300 microns), as well as tryptamine, 5-methoxytryptamine and dopamine, elicited a 3-5 fold increase in glucose transport, as compared with control. This effect was maximal after 90 min, and was concomitant with a 1.8- and 1.5-fold increase in the amounts of glucose transporters GLUT1 and GLUT4 at the cell surface of the cardiomyocytes, as determined by using the photoaffinity label 3H-2-N-[4-(1-azi 2,2,2-trifluoroethyl)benzoyl]-1,3-bis-(D-manno s-4-yl) propyl-2-amine (3H-ATB BMPA). In contrast, 3-3000 microM of the selective 5-HT receptor agonists 5 carboxyamido-tryptamine, alpha-methyl-serotonin, 2-methyl-serotonin or renzapride failed to stimulate glucose transport. The effect of 5-HT was not affected by (i) the 5-HT receptor antagonists methysergide (1 microM), ketanserin (1 microM), cyproheptadine (1 microM), MDL 72222 (1 microM) or ICS 205-930 (3 microM), nor by (ii) the adrenergic receptor antagonists prazosin (1 microM), yohimbine (1 microM) or propranolol (5 microM), nor by (iii) the dopaminergic antagonists SCH 23390 (1 microM) or haloperidol (1 microM). The monoamine oxidase inhibitors clorgyline (1 microM) and tranylcypromine (1 microM) completely suppressed the effect of 5-HT, whereas the control and insulin-stimulated rates of glucose transport were unaffected. Addition of catalase or glutathione diminished the 5 HT-dependent stimulation of glucose transport by 50%; these two factors are known to favour the degradation of H2O2 (which can be formed during the deamination of amines by monoamine oxidases). Glutathione also depressed the stimulatory action of exogenously added H2O2 (20 microM) by 30%. Furthermore, in cells treated with 5_HT, a time-dependent accumulation of 5-hydroxy-1H-indol-3-ylacetic acid (a product of 5-HT metabolism via monoamine oxidases) was observed, which paralleled the changes in glucose transport. In conclusion, the stimulation of glucose transport by 5-HT in cardiomyocytes is not mediated by a 5-HT1, 5-HT2, 5-HT3 or 5 HT4 receptor, nor by an adrenergic or dopaminergic receptor, but is likely to occur through the degradation of by a monoamine oxidase and concomitant formation of H2O2. PMID- 7487899 TI - The 2-chlorodeoxyadenosine-induced cell death signalling pathway in human thymocytes is different from that induced by 2-chloroadenosine. AB - 2-chloroadenosine induced DNA fragmentation and cell death in human thymocytes primarily by Ca(2+)-dependent mechanisms. Incubation of human thymocytes with 2 chlorodeoxyadenosine (5-1000 nM) also induced cell death (apoptosis) which was dependent on macromolecule synthesis and involved activation of an endonuclease which was inhibited by Zn2+. The effect of 2-chlorodeoxyadenosine was prevented by addition of dipyridamole, a strong nucleoside transport inhibitor, or of deoxycytidine, previously shown to compete for uptake by deoxycytidine kinase. 2 Chlorodeoxyadenosine-induced apoptosis did not involve increases in the cytosolic Ca2+ concentration, but required the presence of intracellular Ca2+. It was not inhibited by activators of protein kinase C previously shown to inhibit Ca(2+) dependent cell death. Addition of 2-chlorodeoxyadenosine induced an increase in the amount of p53 in human thymocytes, while 2-chloroadenosine had no effect. These data suggest that 2-chloroadenosine and 2-chlorodeoxyadenosine induce cell death in human thymocytes via different signalling pathways. PMID- 7487900 TI - Agonists for endothelial P2 purinoceptors trigger a signalling pathway producing Ca2+ responses in lymphocytes adherent to endothelial cells. AB - Recirculation of lymphocytes through the body involves their frequent adhesion to endothelial cells but little is known of the signalling pathways between these two cell types. Lymphocytes from patients with chronic lymphocytic leukaemia were loaded with the Ca(2+)-sensitive indicator, fura 2, and allowed to adhere to either glass or monolayers of human umbilical-vein endothelial cells. Addition of ATP or UTP (1-10 microM) to the superfusate produced a transient rise in cytosolic Ca2+ concentration in the lymphocytes adherent to endothelium (24 of 35 cells). In contrast, ATP or UTP (1-10 microM) had no effect on the cytosolic Ca2+ of lymphocytes attached to glass. As the only lymphocyte receptor for ATP (P2Z class) requires higher ATP concentrations ( > 50 microM) for Ca2+ influx and is unresponsive to UTP, the involvement of a lymphocyte P2Z purinoceptor is unlikely. Various agonists including ATP, UTP, 2-methylthioATP, ADP and histamine all stimulated increases in endothelial cytosolic Ca2+ but only ATP and UTP (both agonists for endothelial P2U purinoceptors) triggered Ca2+ transients in adherent lymphocytes. Removal of extracellular Ca2+ did not abolish the ATP-induced rise in cytosolic Ca2+ concentration in lymphocytes adherent to endothelial cells. These findings show that stimulation of endothelial P2U purinoceptors triggers an endothelial-lymphocyte signalling pathway which releases internal Ca2+ in adherent lymphocytes. PMID- 7487902 TI - Isolated rat hepatocytes acquire iron from lactoferrin by endocytosis. AB - The iron-binding protein lactoferrin (Lf) present in blood is metabolized by the liver. Isolated rat hepatocytes vigorously endocytose bovine Lf via recycling Ca2(+)-dependent binding sites, but the uptake of iron from Lf by hepatocytes has not been examined. In this study, isolated rat hepatocytes were incubated with radiolabelled bovine Lf (125I-Lf, 59Fe-Lf or 125I-59Fe-Lf) at 37 degrees C, then washed at 4 degrees C in the presence of dextran sulphate with either Ca2+ or EGTA to distinguish between total bound and internal radioactivity respectively. Cells internalized 125I-Lf protein and Lf-bound 59Fe at maximal endocytic rates of 1700 and 480 mol.cell-1.s-1 respectively. When Lf was normalized for 59Fe content, these endocytic rates were equivalent and reflected an uptake potential of at least 3400 mol of iron.cell-1.s-1. Cells prebound with 125I-59Fe-Lf to Ca2+(-)dependent sites at 4 degrees C internalized more than 80% of both 125I-Lf protein and Lf-bound 59Fe approx. 6 min after warming to 37 degrees C at similar rates (125I-Lf: k(in) = 0.276 min-1, 59Fe: k(in) = 0.303 min-1). Within 4 h at 37 degrees C, cells had released 25% or less internalized Lf protein in the form of acid-soluble 125I-by-products but retained all the Lf-delivered 59Fe. Hyperosmotic disruption of clathrin-dependent endocytosis blocked the uptake of 125I-Lf and Lf-bound 59Fe. Incubation of cells with 125I-59Fe-Lf and a 100 molar excess of diferric transferrin reduced slightly the endocytosis of 125I-Lf protein and 59Fe accumulation. Treatment of cells with the ferric chelator desferrioxamine did not alter uptake of 125I-Lf protein or Lf-bound 59Fe, but the ferrous chelator bathophenanthroline disulphonate slightly elevated endocytosis of 125I-Lf protein and Lf-bound 59Fe. These findings indicate that Lf does not release its bound iron before endocytosis. It was concluded from this study that hepatocytes take up iron from Lf at high rates by a process that requires endocytosis of Lf-iron complexes. PMID- 7487901 TI - Multiple signalling pathways involved in the stimulation of fatty acid and glycogen synthesis by insulin in rat epididymal fat cells. AB - We have investigated the signalling pathways involved in the stimulation of glycogen and fatty acid synthesis by insulin in rat fat cells using wortmannin, an inhibitor of phosphatidylinositol 3-kinase, and rapamycin, which blocks activation of p70 ribosomal S6 protein kinase (p70S6K). Insulin produced a decrease in the activity of glycogen synthase kinase-3 which is likely to be important in the observed stimulation of glycogen synthase. Both of these actions were found to be sensitive to inhibition by wortmannin. Activation of three processes is involved in the stimulation of fatty acid synthesis from glucose by insulin, namely glucose uptake, acetyl-CoA carboxylase and pyruvate dehydrogenase. Whereas wortmannin largely abolished the effects of insulin on glucose utilization and acetyl-CoA carboxylase activity, it was without effect on the stimulation of pyruvate dehydrogenase. Although epidermal growth factor stimulated mitogen-activated protein kinase to a greater extent than insulin, it was unable to mimic the effect of insulin on glycogen synthase, glycogen synthase kinase-3, glucose utilization, acetyl-CoA carboxylase or pyruvate dehydrogenase. Rapamycin also failed to have any appreciable effect on stimulation of these parameters by insulin, although it did block the effect of insulin on p70S6K. We conclude that the activity of phosphatidylinositol 3-kinase is required for the effects of insulin on glycogen synthesis, glucose uptake and acetyl-Co-AN carboxylase, but is not involved in signalling to pyruvate dehydrogenase. Activation of mitogen-activated protein kinase or p70S6K, however, does not appear to be sufficient to bring about the stimulation of fatty acid or glycogen synthesis. Altogether is seems likely that at least four distinct signalling pathways are involved in the effects of insulin on rat fat cells. PMID- 7487903 TI - Uptake of apolipoprotein E-rich and apolipoprotein E-poor subfractions of high density lipoprotein by liver membranes and HepG2 cells. AB - Apolipoprotein (apo) E plays an important role in mediating high-density lipoprotein (HDL) cholesterol transport and uptake by the liver. Evidence for and against the existence of conventional liver receptors for HDL containing apoE have been reported, although the selective uptake of the cholesterol moiety of HDL has been demonstrated. The present study investigated the hepatic uptake of subfractions of HDL separated on the basis of their apoE content. Rabbit HDL and its apoE-rich and apoE-poor subfractions, separated by heparin-Sepharose affinity chromatography, were labelled in their apoprotein moieties with [14C]sucrose and in their cholesteryl ester moiety with 3H. No binding of either subfraction to rabbit liver membranes could be detected. With cultured HepG2 cells, however, there was a high uptake of 3H but a very low uptake of 14C from both HDL subfractions, demonstrating that selective uptake was operating. Addition of unlabelled apoE-poor HDL inhibited the uptake of both labels from the two subfractions to the same extent. These studies, which differed from previously reported investigations by employing native homologous HDL subfractions of known apolipoprotein composition, demonstrated that apoE is not directly involved in the selective uptake of HDL cholesterol by the liver. In the absence of specific binding sites on liver membranes, it is suggested that an alternative mechanism might exist for the clearance of HDL cholesterol from the plasma. PMID- 7487904 TI - Primary structure and tissue-specific expression of blue crab (Callinectes sapidus) metallothionein isoforms. AB - In aquatic animals, synthesis of the metal-binding protein metallothionein (MT) can be induced through exposure to elevated levels of metals in food or water. Whether the different routes of exposure lead to expression of different metallothionein isoforms in different tissues in unknown. In this study we examined the induction of metallothionein isoforms in the hepatopancreas and gills of the blue crab Callinectes sapidus. When blue crabs are exposed to cadmium in their diet, the metal accumulates in the hepatopancreas. Size exclusion and anion-exchange chromatography show the presence of five low molecular-mass cadmium-binding proteins. All of the observed cadmium-binding proteins belong to the class I MT family. They are designated as MT-Ia, MT-Ib, MT Ic, MT-IIa and MT-IIb. All purified proteins run as single peaks upon rechromatography on anion-exchange HPLC, except for MT-Ic, which segregates into two peaks corresponding to MT-Ia and MT-Ic. The amino acid sequence of MT-Ia and MT-Ic is identical. MT-Ib differs from MT-Ia and MT-Ic only in having an extra N terminal methionine. The 18 cysteine residues in MT-Ia and MT-IIa occur in identical positions; however, of the remaining 40 amino acids, 15 are found to be different. MT-IIb is identical with MT-IIa, except for an extra methionine residue at its N-terminal position. It appears therefore that, of the five observed CdMTs, only two are the products of distinct genes. CdMT-Ia and -IIa are posttranslationally modified forms of Ib and IIb, respectively, and CdMT-Ia and Ic appear to be conformational isomers. Cadmium-induced expression of the two genes is tissue-specific. When crabs are exposed to cadmium in water, the metal accumulates in the gills, where it is bound to MT-II. MT-I is virtually absent. PMID- 7487905 TI - Evidence that Asn542 of neprilysin (EC 3.4.24.11) is involved in binding of the P2' residue of substrates and inhibitors. AB - Neprilysin (EC 3.4.24.11) is a Zn2+ metallopeptidase involved in the degradation of biologically active peptides, e.g. enkephalins and atrial natriuretic peptide. The substrate specificity and catalytic activity of neprilysin resemble those of thermolysin, a crystallized bacterial Zn2+ metalloprotease. Despite little overall homology between the primary structures of thermolysin and neprilysin, many of the amino acid residues involved in catalysis, as well as Zn2+ and substrate binding, are highly conserved. Most of the active-site residues of neprilysin have their homologues in thermolysin and have been characterized by site-directed mutagenesis. Furthermore, hydrophobic cluster analysis has revealed some other analogies between the neprilysin and thermolysin sequences [Benchetrit, Bissery, Mornon, Devault, Crine and Roques (1988) Biochemistry 27, 592-596]. According to this analysis the role of Asn542 in the neprilysin active site is analogous to that of Asn112 of thermolysin, which is to bind the substrate. Site-directed mutagenesis was used to change Asn542 to Gly or Gln residues. The effect of these mutations on substrate catalysis and inhibitor binding was examined with a series of thiorphan-like compounds containing various degrees of methylation at the P2' residue. For both mutated enzymes, determination of kinetic parameters with [D-Ala2,Leu5]enkephalin as substrate showed that the large decrease in activity was attributable to an increase in Km (14-16-fold) whereas kcat values were only slightly affected (2-3-fold decrease). This is in agreement with Asn542 being involved in substrate binding rather than directly in catalysis. Finally, the IC50 values for thiorphan and substituted thiorphans strongly suggest that Asn542 of neprilysin binds the substrate on the amino side of the P2' residue by formation of a unique hydrogen bond. PMID- 7487906 TI - Human lymphocyte activation is associated with the early and high-level expression of the endogenous lectin CSL at the cell surface. AB - Lymphocytes undergo activation in response to antigens, cytokines, lectins and antibodies interacting with specific cell-surface molecules or through substances influencing signal transduction pathways. This study shows that human T- and B cells stimulated using phorbol esters or plant lectins express early (2 h using phorbol esters and 24 h using plant lectins) a high level of a polyvalent carbohydrate-binding protein, the cerebellar soluble lectin (CSL), which is in part externalized. The lectin, immunologically related to CDw70, interacts with specific glycoprotein ligands of the lymphocyte surface, including CD3 on T-cells and CD24 on B-cells. Major changes in phosphorylations associated with activation appear as largely CSL-dependent since they are specifically inhibited by anti-CSL Fab fragments. It is suggested that the lectin induces the clustering of specific cell-surface glycoproteins and plays the role of an endogenous amplifier of activation signals. PMID- 7487907 TI - Bradykinin-stimulated phosphatidylcholine hydrolysis in airway smooth muscle: the role of Ca2+ and protein kinase C. AB - The regulation of phosphatidylcholine (PtdCho) hydrolysis by Ca2+ and protein kinase C (PKC) was measured in [3H]palmitate-labelled cultured guinea-pig airway smooth-muscle cells as phosphatidylbutanol ([3H]PtdBut) and phosphatidate ([3H]PtdOH) formation in the presence of butanol. The former is a direct measure of phospholipase D (PLD) activity, whereas the latter, in airway smooth muscle, is indicative of net PtdCho-specific phospholipase C (PLC)-like/diacylglycerol (DG) kinase activity. Bradykinin-stimulated responses exhibited a requirement for extracellular Ca2+ influx, since they were inhibited in the presence of EGTA. This influx was independent of voltage-operated channels, since the L-type channel blocker nifedipine (up to 10 microM) was without effect on bradykinin stimulated responses. In support of this, membrane depolarization with KCl (30 mM) failed to elicit either response. However, bradykinin-stimulated formation of both [3H]PtdBut and [3H]PtdOH was partially inhibited by 100 microM SKF96365. Ionomycin, a Ca2+ ionophore, induced PtdCho hydrolysis to a greater extent than bradykinin, also in an extracellular-Ca(2+)-dependent manner. Thapsigargin induced emptying of intracellular Ca2+ pools elicited the formation of both [3H]PtdBut and [3H]PtdOH and displayed a requirement for extracellular Ca2+. Bradykinin-stimulated PtdCho-specific PLC-like/DG kinase pathway and PLD responses were unaffected by thapsigargin pretreatment, thereby questioning the role of Ins(1,4,5)P3/Ins(1,3,4,5)P4-dependent Ca2+ stores in the receptor stimulation of these activities in airway smooth-muscle cells. In this regard, we have previously demonstrated that the bradykinin-stimulated PtdCho-specific PLD and PLC-like activities can occur under conditions of apparent complete blockade of bradykinin-stimulated Ins(1,4,5)P3 formation by receptor antagonist in guinea pig airway smooth muscle. The PKC inhibitor, Ro31-8220, selectively blocked both bradykinin- and ionomycin-stimulated PLD activity in a concentration-dependent manner (IC50 approx. 1 microM), but was without effect on bradykinin-stimulated PtdCho-PLC-like/DG kinase-derived PtdOH formation. In contrast, an inhibitor of PtdCho-PLC, D609, selectively blocked the formation of [3H]PtdOH in the presence of butanol (PtdCho-PLC-like/DG kinase activity), but not [3H]PtdBut formation. In conclusion, PtdCho hydrolysis appears to occur via two distinguishable routes which both require extracellular Ca2+, whereas only the PLD route is regulated by PKC. PMID- 7487908 TI - Identification of a transcriptional regulatory region of the rat pancreatitis associated protein I (PAP I) gene that confers tissue specificity. AB - We have previously characterized the rat pancreatitis-associated protein I (PAP I) gene by nucleotide sequencing. We describe in this paper its promoter region by analysing the regulatory functions associated with the DNA sequence comprising nt -1253 to + 10 of the gene. That sequence strongly promoted the transcription of the promotorless chloramphenicol acetyltranferase (CAT) gene in cells of pancreatic origin (AR-42J) but not in cells of non-pancreatic origin (Rat 2 and IEC 6). The influence on CAT expression of stepwise 5' deletions in the promoter sequence was monitored in the three cell lines. In pancreatic AR-42J cells, deletion down to position -926 did not affect significantly the expression of the reporter gene. Deletion to nt -685 caused about a 30% decrease in expression. Extending the deletion to nt -444 did not have any additional effect, but a further deletion to nt -180, resulted in a reduction to about 25%. Moreover, deletion from nt -180 to -118 resulted in a further reduction to about one-third of that. Finally, deletion down to nt -61 further reduced activity by a factor of 3, although it remained above background. These results suggest the presence of several positive cis-acting elements in the PAP I promoter. In non-pancreatic cells, CAT expression remained very low when the promoter was deleted down to nt 180. Yet, deletion from -180 to -118 significantly increased CAT expression, suggesting suppression of a negative cis-acting element. Further deletion down to nt -61 decreased CAT activity by a factor of 5. The region between nt -180 and 61 was subjected to footprint analysis. A similar pattern of DNase protection was obtained with AR-42J and Rat 2 nuclear extracts, the only protected region extending from nt -125 to -95. That region was further analysed by inserting the nt -180 to -81 fragment, in both orientations, upstream of thymidine kinase (TK) or simian virus 40 (SV40) promoter-CAT constructs. In all cases CAT expression was increased in pancreatic cells but reduced in Rat 2 cells. These results indicated the presence of cell-specific positive and negative elements within that region. PMID- 7487909 TI - Role of cyclic GMP in the control of capacitative Ca2+ entry in rat pancreatic acinar cells. AB - We have investigated the possible roles of cyclic GMP (cGMP) in initiating or regulating capacitiative Ca2+ entry in rat pancreatic acinar cells. In medium containing 1.8 mM external Ca2+, thapsigargin activated Ca2+ entry and slightly but significantly increased intracellular cGMP concentration. This rise in cGMP levels was prevented by pretreating the cells with the guanylate cyclase inhibitor, LY-83583, or by omitting Ca2+ during stimulation by thapsigargin or methacholine. LY-83583 and NG-nitro-L-arginine (L-NA, an inhibitor of NO synthase) both had a small inhibitory effect on Ca2+ entry when they were added after thapsigargin in Ca2(+)-containing medium, and they reduced by 32 and 48% respectively the thapsigargin-induced capacitative Ca2+ entry when added to the cells during a 20 min preincubation period. However, neither dibutyryl cGMP (Bt2cGMP) nor sodium nitroprusside, an NO mimic, affected either basal intracellular Ca2+ concentration [Ca2+]i or thapsigargin-induced capacitative Ca2+ entry. Further, the inhibitory effects observed after preincubation with LY 83583 or L-NA could not be prevented by preincubation with Bt2cGMP, nor could they be reversed by adding Bt2cGMP, 8-bromo-cGMP or sodium nitroprusside acutely after activation of capacitative Ca2+ entry by thapsigargin. Finally, pretreatment of cells with LY-83583 or L-NA did not affect Ca2+ signalling in response to 1 microM methacholine, including the pattern of [Ca2+]i oscillations. In conclusion, in pancreatic acinar cells, the rise in cellular cGMP levels appears to depend on, rather than cause, the increase in [Ca2+]i with agonist stimulation. PMID- 7487910 TI - Localization of rat endothelin-converting enzyme to vascular endothelial cells and some secretory cells. AB - Endothelin is a potent vasoconstrictive peptide that is produced by vascular endothelial cells; it is formed from its precursor, big endothelin, by endothelin converting enzyme (ECE). In this work, ECE was studied using specific monoclonal antibodies. In immunoblotting, ECE was estimated to be a 300 kDa protein on SDS/PAGE under non-reducing conditions, and 130 kDa under reducing conditions. Cross-linking experiments revealed that ECE is composed of two disulphide-linked subunits. Localization of ECE was studied at the cellular and subcellular levels in various rat tissues and cells. High-level expression of ECE was observed in membrane fractions of simian virus 40-transformed rat endothelial cells by immunoblotting, but the immunoreactive band was absent form aortic smooth muscle cells and cytosolic fractions of endothelial cells. In immunohistochemical analysis, ECE was found to be localized in the endothelial cells of the aorta, lung, kidney, liver and heart. Confocal immunofluorescent microscopy showed that most of the ECE in endothelial cells and cells transfected with ECE cDNA was clustered along the plasma membrane. Intact COS or CHO cells transfected with ECE cDNA rapidly and efficiently cleaved big endothelin-1 added to the culture medium. Thus endothelial cells express ECE on the plasma membrane and the active site of the enzyme faces outside the cells, i.e. it is an ectoenzyme. Other than endothelial cells, ECE was also present in some secretory cells. The enzyme was abundant in the adrenal gland, and localized in chromaffin cells. ECE was also highly condensed in pancreatic islet beta cells. It is concluded that ECE and endothelin may be involved in the regulated secretion of hormones. PMID- 7487911 TI - The lysosomal membrane glycoproteins Lamp-1 and Lamp-2 are present in mobilizable organelles, but are absent from the azurophil granules of human neutrophils. AB - The subcellular localization of two members of a highly glycosylated protein group present in lysosomal membranes in most cells, the lysosome-associated membrane proteins 1 and 2 (Lamp-1 and Lamp-2), was examined in human neutrophil granulocytes. Antibodies that were raised against purified Lamp-1 adn Lamp-2 gave a distinct granular staining of the cytoplasm upon immunostaining of neutrophils. Subcellular fractionation was used to separate the azurophil and specific granules from a light-membrane fraction containing plasma membranes and secretory vesicles, and Western blotting was used to determine the presence of the Lamps in these fractions. The results show that Lamp-1 and Lamp-2 are present in the specific-granule-enriched fraction and in the light-membrane fraction, but not in the azurophil granules. Separation of secretory vesicles from plasma membranes disclosed that the light-membrane Lamps were present primarily in the secretory vesicle-enriched fraction. During phagocytosis both Lamp-1 and Lamp-2 became markedly concentrated around the ingested particle and they both appear on the cell surface when the secretory organelles are mobilized. PMID- 7487912 TI - Cloning of the cDNA encoding a novel rat mast-cell proteinase, rMCP-3, and its expression in comparison with other rat mast-cell proteinases. AB - A cDNA encoding a novel rat mast-cell proteinase (MCP) named rMCP-3 was successfully cloned and sequenced from the peritoneal cells of Lewis rats infected with the intestinal nematode Nippostrongylus brasiliensis by using the combination of reverse transcription-PCR and rapid-amplification-of-cDNA-ends ('RACE') methods. The cDNA was 979 bp long and included a 741 bp open reading frame. When the deduced amino acid sequence was compared with those of other known mast-cell proteinases, rMCP-3 was considered to be translated as a preproenzyme with a 19-amino-acid signal peptide, a two-amino-acid activation peptide and a 226-amino-acid mature enzyme. The amino acid identity in the mature enzyme was 52.9% and 55.1% with rMCP-1 and rMCP-2 respectively. The rMCP-3 mRNA was not detected in the peritoneal cells of mast-cell-deficient Ws/Ws rats, though it was strongly detected in those of littermate +/+ and Lewis rats, indicating the mast-cell origin of rMCP-3 In addition to being present in peritoneal mast cells, the rMCP-3 mRNA was strongly detected in the skin, tongue, and RBL2H3 rat basophilic leukaemia cells and weakly in the jejunum of N. brasiliensis-infected rats by RNA blot analysis using a rMCP-3 gene-specific probe. By reverse transcription-PCR, the rMCP-3 mRNA was also detected in the lung. While the expression of rMCP-1 and rMCP-2 are clearly restricted in connective-tissue mast cells and mucosal mast cells respectively, rMCP-3 was widely expressed in both types of mast cells with a predominance in connective tissue mast cells. PMID- 7487913 TI - Dependence of in vivo glutamine synthetase activity on ammonia concentration in rat brain studied by 1H - 15N heteronuclear multiple-quantum coherence-transfer NMR. AB - The dependence of the in vivo rate of glutamine synthesis on the substrate ammonia concentration was studied in rat brain by 1H-15N heteronuclear multiple quantum coherence-transfer NMR in combination with biochemical techniques. In vivo rates were measured at various steady-state blood and brain ammonia concentrations within the ranges 0.4-0.55 mumol/g and 0.86-0.98 mumol/g respectively, after low-rate intravenous 15NH4+ infusion (isotope chase). The rate of glutamine synthesis at steady state was determined from the change in brain [5-15N]glutamine levels during isotope chase, observed selectively through the amide proton by NMR, and 15N enrichments of brain glutamine and of blood and brain ammonia measured byN gas chromatography-MS. The in vivo rate (v) was 3.3 4.5 mumol/h per g of brain at blood ammonia concentrations (s) of 0.40-0.55 mumol/g. A linear increase of 1/v with 1/s permitted estimation of the in vivo glutamine synthetase (GS) activity at a physiological blood ammonia concentration to be 0.4-2.1 mumol/h per g. The observed ammonia-dependence strongly suggests that, under physiological conditions, in vivo GS activity is kinetically limited by sub-optimal in situ concentrations of ammonia as well as glutamate and ATP. Comparison of the observed in vivo GS activity with the reported in vivo rates of glutaminase and of gamma-aminobutyrate (GABA) synthesis suggests that, under mildly hyperammonaemic conditions, glutamine is synthesized at a sufficiently high rate to serve as a precursor of GABA, but glutaminase-catalysed hydrolysis of glutamine is too slow to be the sole provider of glutamate used for GABA synthesis. PMID- 7487914 TI - Polyenoic very-long-chain fatty acids mobilize intracellular calcium from a thapsigargin-insensitive pool in human neutrophils. The relationship between Ca2+ mobilization and superoxide production induced by long- and very-long-chain fatty acids. AB - Fatty acids with more than 22 carbon atoms (very-long-chain fatty acids; VLCFAs) are normal cellular components that have been implicated in the pathophysiology of a number of peroxisomal disorders. To date, however, essentially nothing is known regarding their biological activities. Ca2+ mobilization is an important intracellular signalling system for a variety of agonists and cell types. Given that several polyunsaturated long-chain fatty acids mobilize intracellular Ca2+ and that we have postulated that the VLCFAs may be involved in signal transduction, we examined whether the tetraenoic VLCFA induced Ca2+ mobilization in human neutrophils. We report that fatty acid-induced intracellular Ca2+ mobilization declined for fatty acid species of more than 20 carbon atoms, but increased again as the carbon chain length approached 30. This Ca2+ mobilization occurred independently of inositol 1,4,5-triphosphate production and protein kinase C translocation and involved both the release of Ca2+ from the intracellular stores and changes to the influx or efflux of the ion. We further observed that triacontatetraenoic acid [30:4(n-6)] mobilized Ca2+ from a thapsigargin-insensitive intracellular pool distinct from the thapsigargin sensitive pools affected by arachidonic acid [20:4(n - 6)] or N-formyl-L methionyl-L-leucyl-L-phenylalanine (fMLP). 20:4 (n - 6) induced strong superoxide production (chemiluminescence) which was inhibited by thapsigargin pretreatment. In contrast, fatty acid-induced superoxide production progressively declined as the carbon chain length increased beyond 20-22 carbon atoms. Further studies suggested that the thapsigargin-insensitive Ca2+ mobilization elicited by 30:4 (n - 6) was not related to oxyradical formation, while the thapsigargin-sensitive Ca2+ mobilization induced by 20:4 (n - 6) may be involved in the initiation but not necessarily the maintenance of superoxide production. In conclusion, this is the first report to demonstrate a biological activity for the VLCFA and indicates that 30:4 (n - 6) influences second messenger systems in intact cells that differ from those affected by long-chain fatty acids such as 20:4 (n - 6). PMID- 7487915 TI - Analysis of the structural features of the C-terminus of GLUT1 that are required for transport catalytic activity. AB - C-terminally truncated and mutated forms of GLUT1 have been constructed to determine the minimum structure at the C-terminus required for glucose transport activity and ligand binding at the outer and inner binding sites. Four truncated mutants have been constructed (CTD24 to CTD27) in which 24 to 27 amino acids are deleted. In addition, point substitutions of R468-->L, F467-->L and G466-->E have been produced. Chinese hamster ovary clones which were transfected with these mutant GLUT1s were shown, by Western blotting and cell-surface carbohydrate labelling, to have expression levels which were comparable with the wild-type clone. Wild-type levels of 2-deoxy-D-glucose transport activity were retained only in the clone transfected with the construct in which 24 amino acids were deleted (CTD24). The CTD25, CTD26 and CTD27 clones showed markedly reduced transport activity. From a kinetic comparison of the CTD24 and CTD26 clones it was found that the reduced transport was mainly associated with a reduced Vmax. value for 2-deoxy-D-glucose uptake but with a slight lowering of the Km. These data establish that the 24 amino acids at the C-terminus of GLUT1 are not required for the transport catalysis. However, the point mutations of F467L and G466E (26 and 27 residues from the C-terminus) did not significantly perturb the kinetics of 2-deoxy-D-glucose transport. The substitution of R468L produced a slight, but significant, lowering of the Km. The ability of the truncated GLUt1s to bind the exofacial ligand, 2-N-4-(1-zai-2,2,2-trifluoroethyl)benzoyl-1,3-bis (D-mannos- 4-yl-oxy) -2-propylamine (ATB-BMPA), and the endofacial ligand, cytochalasin B, were assessed by photolabelling procedures. The ability to bind ATB-BMPA was retained only in the CTD24 truncated mutant and was reduced to levels comparable with those of the non-transfected clone in the other mutant clones. Cytochalasin B labelling was unimpaired in all four mutated GLUT1s. These data establish that a minimum structure at the C-terminus of GLUT1, which is required for the conformational change to expose the exofacial site, includes amino acids at positions Phe-467 and Arg-468; however, these amino acids are not individually essential. PMID- 7487916 TI - Evidence for substrate-cycling of 3-, 3,4-, 4-, and 4,5-phosphorylated phosphatidylinositols in plants. AB - Short-term 32P labelling and enzymic dissection of inositol phospholipids was used to study the turnover of 3-, 3,4-, 4-, and 4,5-phosphorylated phosphatidylinositols in the plant Spirodela polyrhiza L. Analysis of label in the whole headgroup reveals that phosphatidylinositol 3- and 4-monophosphates (PtdIns3P and PtdIns4P) and phosphatidylinositol 3,4- and 4,5-bisphosphates [PtdIns(3,4)P2 and PtdIns(4,5)P2] all turn over with a half-life of approximately 2-5 h. Analysis of the labelling of individual phosphomonoesters and phosphodiesters of these lipids indicates a rapid equilibration of label between the 4- and 5-monoester phosphates of PtdIns(4,5)P2 within 5 h and largely independent of changes of labelling in the diester. We observed substantially slower equilibration of label (within approximately 27 h) between the monoester and diester of PtdIns4P. These studies therefore indicate that PtdIns4P and PtdIns(4,5)P2 participate in substrate-cycling reactions, evidence for which has been described experimentally only in erythrocytes, and give confirmation in vivo of the previous detection of inositol phospholipid phosphomonoesterase activity. Similar analyses of label in PtdIns3P and PtdIns(3,4)P2 reveal the likely participation of these molecules in substrate cycles and hence for the first time the presence of PtdIns3P 3-phosphatase and PtdIns(3,4)P2 4-phosphatase activities in plants. PtdIns3P and PtdIns(3,4)P2 undergo turnover at rates similar to those of PtdIns4P and PtdIns(4,5)P2. Estimates are made of the relative sizes of the pools of phospholipid participating in the turnover process. PMID- 7487917 TI - Preparation and characterization of a D-myo-inositol 1,4,5-trisphosphate-specific antibody. AB - Antibodies against Ins(1,4,5)P3 were raised by immunizing rabbits with two types of InsP3-BSA conjugates which were synthesized by covalently coupling Ins(1,4,5)P3 to the carrier protein via alkyl linkages. The anti-Ins(1,4,5)P3 antibody was detected by a novel ELISA using Ins(1,4,5)P3-immobilized microtitre plates. Both antiserum preparations showed specific binding with Ins(1,4,5)P3, with titres of 1:4000. Most inositol phosphates, including Ins1P, Ins(4,5)P2, Ins(1,3,4)P3, Ins(1,5,6)P3, Ins(1,2,5,6)P1, Ins(3,4,5,6)P4, Ins(1,3,4,5,6)P5, InsP6, and PtdIns(4,5)P2, did not exhibit significant molecular interactions with the antibodies. Ins(1,3,4,5)P4, however, cross-reacted with these antibodies with one-third of the affinity as that of Ins(1,4,5)P3, in part due to the largely shared structural motifs. The differential affinity was significantly improved by affinity purification on Ins(1,4,5)P3-agarose. The affinity-purified antibody displayed IC50 values of 12 nM and 730 nM for Ins(1,4,5)P3 and Ins(1,3,4,5)P4 respectively, according to a competitive ELISA; these values are in line with those reported for the Ins(1,4,5)P3 receptor. The modes of ligand recognition at the binding sites of these two types of biomolecules are, however, different. Moreover, although the ligand binding was interfered with by multivalent anions such as ATP4-, HPO4(3-) and SO4(2-) at high concentrations, no inhibition was noted with heparin, an antagonist of the Ins(1,4,5)P3 receptor. PMID- 7487918 TI - Factors affecting the stereospecificity and catalytic efficiency of the tryptophan synthase-catalysed exchange of the pro-2R and pro-2S protons of glycine. AB - 13C-NMR has been used to follow the tryptophan synthase (EC 4.2.1.20)-catalysed hydrogen-deuterium exchange of the pro-2R and pro-2S protons of [2-13C]glycine. The first- and second-order rate constants for exchange when the alpha 2 beta 2 enzyme complex is or is not saturated with glycine have been determined at pH 7.0 and 7.8. At pH 7.8 the effects of binding the allosteric effector, DL-alpha glycerol 3-phosphate, and of removing the alpha-subunits have been examined. The beta-subunits preferentially catalyse the exchange of the pro-2R proton of glycine, but adding alpha-subunits decreases the stereospecificity of the exchange reactions. Likewise, binding of DL-alpha-glycerol 3-phosphate to the alpha 2 beta 2 enzyme complex causes a further decrease in the stereospecificity of this reaction. The stereospecificity of the second-order exchange reaction catalysed by the beta-subunits is 136-fold larger than that of the alpha 2 beta 2 enzyme complex in the presence of DL-alpha-glycerol 3-phosphate, while there is only a 5-fold decrease in the stereospecificity of the first-order exchange reaction under the same conditions. We discuss how these results relate to current theories which attempt to explain how the alpha-subunits and DL-alpha glycerol 3-phosphate modify the catalytic properties of tryptophan synthase. PMID- 7487919 TI - The Gly-54-->Asp allelic form of human mannose-binding protein (MBP) fails to bind MBP-associated serine protease. AB - The human mannose-binding protein (MBP) is a pattern recognition molecule that appears to play a role in initial host defence. MBP activates the complement cascade and it may act as an opsonin both in the absence and in the presence of complement. A number of distinct MBP allelic forms exist in different population groups. An allele that occurs in 5-7% of Caucasians was identified by an inability to activate the complement system. A homozygous mutation at base pair 230 of the MBP gene results in a Gly-to-Asp substitution at the fifth collagen repeat. It appears that the resultant protein, MBPD, is able to form high-order multimers that bind bacteria but do not support complement activation. Recently a novel serine protease, the MBP-associated serine protease (MASP), has been described. MBP-MASP complexes circulate in serum and result in the direct activation of a novel complement pathway (lectin pathway) in the absence of the first complement components. In this study we demonstrate that MASP and its proenzyme proMASP are unable to bind to recombinant (r)MBPD. This lack of a MASP rMBPD association corresponds to a failure of the Gly-54-->Asp form of MBP to activate complement. Our results provide a biochemical basis for the functional deficit in the Gly-54-->Asp allelic form of MBP and suggest that the proMASP/MASP binding site maps to the fifth collagen repeat of MBP. PMID- 7487920 TI - Phosphonate inhibitors of protein-tyrosine and serine/threonine phosphatases. AB - In all, 15 aryl-containing phosphonates have been synthesized and tested for their effect on protein-tyrosine phosphatase (PTPase) activity. Two compounds, (naphth-2-yl) difluoromethylphosphonic acid (12) and (napthy-1-yl) difluoromethylphosphonic acid (13) have been found to inhibit dephosphorylation of [32P]insulin receptors by PTP-1B, a protein tyrosine phosphatase (PTPase), with IC50 values of 40-50 microM. Compound 12 competitively inhibited insulin receptor dephosphorylation by PTP-1B. Compound 12 also inhibited PTP-1B-catalysed dephosphorylation of a synthetic tyrosine phosphorylated substrate poly(Glu80 Tyr20) at the same potency, indicating that 12 acted via interaction with the PTPase. Additionally, 12 inhibited insulin-receptor PTPase(s) and epridermal growth-factor-receptor PTPase(s) present in solubilized membranes from CHO (Chinese-hamster ovary)/HIRc and A431 cells respectively. IC50 values of 40-50 microM were obtained in all cases with compound 12. Of note is the fact that these compounds did not have any effect on insulin-receptor autophosphorylation. Nine out of the 15 compounds potently inhibited serine/threonine phosphatase PP 2A activity without any effect on serine/threonine phosphatase PP-1 when tested at a concentration as high as 675 microM. The most potent compounds acting toward PP-2A had IC50 values of 45-50 microM. These PP-2A inhibitors could be useful tools for studying serine/threonine-phosphatase-mediated signal transduction. Two compounds, 12 and 13, inhibited both tyrosine phosphatase PTP-1B and serine/threonine phosphatase PP-2A with similar potency; IC50 values being 40-50 microM in both cases. Details of the synthesis of compounds 10, 11 and 13 are given in Supplementary Publication SUP 50177 (6 pages), which has been deposited at the British Library Document Supply Centre, Boston Spa, Wetherby, West Yorkshire LS23 7BQ, U.K., from whom copies can be obtained on the terms indicated in Biochem. J. (1995) 305, 9. PMID- 7487921 TI - Ionic regulation of endonuclease activity in PC12 cells. AB - We have investigated the Ca2+ dependency of DNA degradation into nucleosome-sized fragments in intact chromaffin-like PC12 cells and PC12 nuclear fractions. In intact cells we were unable to trigger DNA fragmentation by inducing either transient or sustained elevations of cytoplasmic Ca2+ ([Ca2+]i) with the Ca2+ ionophore ionomycin. On the contrary, DNA fragmentation was induced in intact cells by the intracellular Zn2+ chelator NNN'N'-tetrakis-(2 pyridylmethyl)ethylenediamine (TPEN). To characterize further PC12 cell endonuclease activity, we then investigated digestion by purified PC12 cell fractions of exogenously added plasmids. In nuclear fractions two endonuclease activities were identified: an acidic (pH 5.0) endonuclease activity that was fully Ca2+- and Mg(2+)-independent; and a neutral (pH 7.6) endonuclease activity that was Ca(2+)-independent but Mg(2+)-dependent. Both endonuclease activities were inhibited by Zn2+. Nuclear membrane permeabilization greatly enhanced plasmid digestion at pH 7.6, but not at pH 5.0. This suggests that neutral endonuclease was located in a membrane-bound compartment, whereas acidic endonuclease was freely accessible to the substrate even in the presence of an intact nuclear membrane. In intact nuclei, digestion of genomic DNA could not be triggered by increasing the bivalent cation composition of the medium. On the contrary, in hypotonic medium we observed a large spontaneous nucleolytic DNA degradation that was increased by Zn2+ chelation. However, an acidic pH shift was a potent stimulus for DNA fragmentation in isotonic as well as hypotonic medium. PMID- 7487923 TI - Human diadenosine 5',5"'-P1,P4-tetraphosphate pyrophosphohydrolase is a member of the MutT family of nucleotide pyrophosphatases. AB - The cDNA and derived amino acid sequence of human diadenosine 5',5"'-P1,P4 tetraphosphate pyrophosphohydrolase have been determined with the aid of the GenBank Expressed Sequence Tag database. This enzyme possesses a modification of the MutT sequence motif found in certain nucleotide pyrophosphatases. It is unrelated to the enzymes of diadenosine tetraphosphate catabolism found in prokaryotes and fungi. PMID- 7487924 TI - Induction of apoptosis by excessive polyamine accumulation in ornithine decarboxylase-overproducing L1210 cells. AB - Deregulation of polyamine transport in L1210 cells overexpressing ornithine decarboxylase leads to a lethal accumulation of spermidine. We now provide evidence that over-accumulation of natural and synthetic polyamines, but not putrescine, rapidly induces apoptosis, as shown by hypercondensation of peripheral chromatin and internucleosomal cleavage, followed by nuclear fragmentation. Polyamine oxidation is not responsible for the apoptosis observed. Thus, abnormally high polyamine pools could be an important physiological trigger of apoptosis. PMID- 7487922 TI - The regulation of protein transport to the nucleus by phosphorylation. PMID- 7487925 TI - Acute cold-induced suppression of ob (obese) gene expression in white adipose tissue of mice: mediation by the sympathetic system. AB - The effect of acute exposure to cold on the expression of the ob (obese) gene, which encodes a protein that plays a critical role in the regulation of energy balance and body weight, has been examined in epididymal white adipose tissue of mice. Overnight (18 h) exposure of mice to a temperature of 4 degrees C led to the disappearance of ob mRNA in epididymal white fat, and subsequent studies showed that a cold-induced loss of ob mRNA could occur in as little as 2-4 h of exposure to 4 degrees C. When mice exposed to cold for 18 h were returned to the warm (24 degrees C), there was a rapid stimulation of the expression of the ob gene, the mRNA returning within 2.5 h. Administration of noradrenaline led to a reduction in the level of ob mRNA in mice maintained in the warm, while isoprenaline resulted in the disappearance of the mRNA; these changes in ob mRNA were paralleled by similar changes in lipoprotein lipase mRNA. In contrast to white fat, the level of lipoprotein lipase mRNA in brown adipose tissue was increased by noradrenaline and isoprenaline. It is concluded that there is a cold induced suppression of ob gene expression in white adipose tissue of mice and that this is mediated primarily by the sympathetic system. The profound effect of cold on ob gene expression indicates that the ob system relates to energy expenditure, as well as to satiety. PMID- 7487927 TI - Characterization of rat spleen prostaglandin H D-isomerase as a sigma-class GSH transferase. AB - The prostaglandin H D-isomerase of rat immune accessory cells has been purified from spleen by a simple procedure, and its high specificity and activity [Urade, Fujimoto, Ujihara and Hayaishi (1987) J. Biol. Chem. 262, 3820-3825] have been confirmed in an assay coupled to prostaglandin H synthase. The enzyme also decreases the formation of 12[S]-hydroxy-5,8,10-heptadecatrienoic acid formed by the synthase in the presence of GSH and increases the overall rate of arachidonate oxidation. A partial amino acid sequence shows a strong relationship to GSH transferases of parasitic helminths and molluscs, indicating that it is the first example of a vertebrate sigma-class GSH transferase, and suggesting that certain helminth GSH transferases may be involved in prostaglandin synthesis. PMID- 7487926 TI - The activation of distinct mitogen-activated protein kinase cascades is required for the stimulation of 2-deoxyglucose uptake by interleukin-1 and insulin-like growth factor-1 in KB cells. AB - The uptake of 2-deoxyglucose into KB cells was stimulated about 2-fold by interleukin-1 (IL1), anisomycin or insulin-like growth factor-1 (IGF1). Stimulation by IL1 and anisomycin was prevented by SB 203580, a specific inhibitor of the mitogen-activated protein (MAP) kinase homologue termed 're activating kinase' [RK; also known as p38, p40 and CSBP (cytokine synthesis anti inflammatory-drug-binding protein)], but was unaffected by PD 98059, a specific inhibitor of the activation of the classical MAP kinase pathway. In contrast, the stimulation of 2-deoxyglucose uptake by IGF1 was blocked by PD 98059 and unaffected by SB 203580. Consistent with these observations, IL1 and anisomycin were potent activators of MAP kinase-activated protein (MAPKAP) kinase-2, a physiological substrate of RK, whereas IGF1 was only a very weak activator of MAPKAP kinase-2. Conversely, IGF1 was a stronger activator of p42 MAP kinase than IL1 or anisomycin. These results imply that the activation of distinct MAP kinase pathways is required for the stimulation of glucose transport by IL1/anisomycin and IGF1 in KB cells, and suggest that the combined use of SB 203580 and PD 98059 is a powerful new approach to explore the roles of different MAP kinase cascades in cell regulation. PMID- 7487929 TI - Factor IX Zutphen: a Cys18-->Arg mutation results in formation of a heterodimer with alpha 1-microglobulin and the inability to form a calcium-induced conformation. AB - Factor IX Zutphen is a variant factor IX molecule isolated from the blood of a patient with severe haemophilia B. The molecular defect in factor IX Zutphen is a Cys18-->Arg mutation as a result of a T-->C transition at residue 6427 of the factor IX gene of the patient. The mutation disrupts the disulphide bond in the Gla-domain between Cys18 and Cys23. The remaining free cysteine residue results in the formation of a 95 kDa complex with alpha 1-microglobulin through an intermolecular disulphide bond. The same complex circulates at high levels in plasma of carriers of the mutation. The variant molecule has a calcium-binding defect, which is shown not to be caused by incomplete gamma-carboxylation. Factor IX Zutphen can not bind to phospholipids and can not be activated by factor XIa or by factor VIIa-tissue factor complex. Two sequential metal ion-dependent conformational transitions (factor IX-->factor IX'-->factor IX*) have been proposed for human factor IX [Liebman (1987) J. Biol. Chem. 262, 7605-7612], based upon the metal ion requirements for binding to anti-factor IX:Mg(II) antibodies, which are specific for the factor IX' conformation, and anti-factor IX:Ca(II) antibodies, which are specific for the factor IX* conformation. We used these conformation-specific antibodies, and antibodies raised against a synthetic peptide corresponding to residues 35-50 of human factor IX [anti-factor IX(35 50)] to study the metal ion-induced conformation of factor IX Zutphen. The disruption of the disulphide bond in the Gla-domain, maybe in combination with the complex with alpha 1-microglobulin, destabilized the factor IX' conformation. The formation of the factor IX* conformation was prevented independent of the presence of alpha 1-microglobulin. The disulphide bond in the Gla-domain is therefore essential for the calcium-dependent conformation and function of factor IX. PMID- 7487928 TI - Identification of the integrin alpha 3 beta 1 as a component of a partially purified A-system amino acid transporter from Ehrlich cell plasma membranes. AB - We have previously reported [McCormick and Johnstone (1988) Proc. Natl. Acad. Sci. U.S.A. 85, 7877-7881] the partial purification of the Na(+)-dependent A system amino acid transporter from Ehrlich cell plasma membranes and have suggested that a 120-130 kDa peptide, a major component of the purified fraction [octyl glucoside (OG) extract], is involved in Na(+)-dependent amino acid transport. In the present study, N-terminal sequence analysis of the 120-130 kDa peptide revealed a sequence similar to that of the alpha 3 subunit of the integrin alpha 3 beta 1. The presence of alpha 3 beta 1 was confirmed by Western blots of the OG extract probed with anti-alpha 3 or -beta 1 antibodies. Western blots also showed that an antibody originally raised against the 120-130 kDa peptide crossreacts with both the alpha 3 and beta 1 integrin subunits. Co purification of alpha 3 beta 1 and Na(+)-dependent transport activity suggested that the two activities might be associated. Evidence that alpha 3 plays a role in transport is shown by the fact that an antibody against human alpha 3, but not beta 1, removed transport activity (approximately 25% loss) from cholate solubilized Ehrlich membranes. Further purification of OG extracts using concanavalin A and wheat-germ lectin columns resulted in the separation of transport activity from the bulk (but not all) of alpha 3 beta 1 integrin without loss of the transport activity. These results indicate that the integrin itself is not essential for amino acid transport. Reconstitution of a purified alpha 3 beta 1-depleted protein fraction showed high levels of Na(+)-dependent, alpha methylaminoisobutyric-acid-inhibitable amino acid transport in proteoliposomes, whereas reconstituted integrin alone showed little transport activity. However, in the integrin-depleted fractions, high amino acid uptake occurred in K+ which compromised the accurate measurement of the Na(+)-dependent component of uptake. The data suggest that alpha 3 may be associated with the A-system transporter and may modulate the activity of this carrier. Moreover, transfection of K562 and RD cells with human alpha 3 and alpha 2 cDNA showed that the former but not the latter increased A-system transport, thus providing more direct evidence that alpha 3 may modulate A-system transport activity. PMID- 7487930 TI - In search of a selective inhibitor of the induced transport of small solutes in Plasmodium falciparum-infected erythrocytes: effects of arylaminobenzoates. AB - Following invasion of the human erythrocyte by the malaria parasite, Plasmodium falciparum, there appear in the parasitized cell new, high-capacity permeation pathways that transport a diverse range of low-molecular-mass solutes. In this study a series of 16 arylaminobenzoates, analogues of the Cl- channel blocker 5 nitro-2-(3-phenylpropylamino)benzoic acid (NPPB), were tested for their effects on the transport of choline, a univalent cation, into malaria-infected cells. A number of the arylaminobenzoates were found to be potent inhibitors of malaria induced choline transport and to be similarly effective at blocking the induced transport of the uncharged pyrimidine nucleoside thymidine and the univalent anion lactate. The data are consistent with the hypothesis that much of the induced transport of cations, anions and non-electrolytes into parasitized cells is via broad-specificity, anion-selective pathways of a single type. A comparison of the effects of the arylaminobenzoates on malaria-induced transport with their effects on a number of representative anion transport systems in normal mammalian cells suggests that it is possible to identify pharmacological agents that block the malaria-induced pathway while not significantly affecting important transport mechanisms in host tissues. The most potent of the induced-transport inhibitors identified were shown to inhibit [3H]hypoxanthine incorporation in in vitro parasite growth assays. These data support the view that the induced-transport pathway may be a viable pharmacological target. PMID- 7487931 TI - Transcriptional activation of the H-ferritin gene in differentiated Caco-2 cells parallels a change in the activity of the nuclear factor Bbf. AB - In this paper, we examine the mechanisms that regulate the expression of the heavy (H) ferritin subunit in the colon carcinoma Caco-2 cell line allowed to differentiate spontaneously in vitro. The differentiation process of these cells in continuous culture is accompanied by an accumulation of the mRNA coding for the apoferritin H chain. The analysis of Caco-2 subclones stably transfected with an H-chain promoter-chloramphenicol acetyltransferase (CAT) construct revealed that the mRNA increase is paralleled by an enhanced transcription of the H gene, driven by the -100 to +4 region of the H promoter. The H gene transcriptional activation seems to be a specific feature of differentiated Caco-2 cells, since the activity of other promoters did not change upon differentiation. The -100 to +4 region of the H promoter binds a transcription factor called Bbf (B-box binding factor); electrophoretic-mobility-shift-assay analyses showed that the retarded complex due to Bbf-H promoter interaction is significantly increased in the differentiated cells. We propose that the activation of H-ferritin gene expression may be associated with the establishment of a differentiated phenotype in Caco-2 cells, and that the H-ferritin gene transcriptional up-regulation is accompanied by a modification in the activity of the transcription factor Bbf. PMID- 7487932 TI - Mouse gastric mucin: cloning and chromosomal localization. AB - Mucins protect gastric epithelium by maintaining a favourable pH gradient and preventing autodigestion. The purpose of this study was to clone a mouse gastric mucin which would provide a foundation for analysis of mucin gene regulation. Mucin was purified from the glandular portion of gastric specimens and deglycosylated by HF solvolysis. Antibodies against native and deglycosylated mouse gastric mucin (MGM) were raised in chickens. Screening of a mouse stomach cDNA library with the anti-(deglycosylated MGM) antibody yielded partial clones containing a 48 bp tandem repeat and 768 bp of non-repetitive sequence. The 16 amino-acid tandem repeat has a consensus sequence of QTSSPNTGKTSTISTT with 25% serine and 38% threonine. The MGM tandem repeat sequence bears no similarity to previously identified mucins. The MGM non-repetitive region shares sequence similarity with human MUC5AC and, to a lesser extent, human MUC2 and rat intestinal mucin. Northern blot analysis reveals a polydisperse message beginning at 13.5 kb in mouse stomach with no expression in oesophagus, trachea, small intestine, large intestine, caecum, lung or kidney. Immunoreactivity of antibodies against deglycosylated MGM and against a synthetic MGM tandem repeat peptide was restricted to superficial mucous cells, antral glands and Brunner's glands in the pyloric-duodenal region. DNA analysis shows that MGM recognizes mouse and rat DNA but not hamster, rabbit or human DNA. The MGM gene maps to a site on mouse chromosome 7 homologous to the location of a human secretory mucin gene cluster on human chromosome 11p15. Due to sequence similarity and predominant expression in the stomach, the MGM gene may be considered a MUC5AC homologue and named Muc5ac. PMID- 7487933 TI - Chloroquine augments the binding of insulin to its receptor. AB - The effect of chloroquine on the interaction of insulin with its receptor has been investigated under both equilibrium and non-equilibrium conditions. Chloroquine was found to augment insulin binding in a pH-dependent manner between pH 6.0 and pH 8.5, with the maximum occurring at approximately pH 7.0. Analysis of the equilibrium binding data in terms of independent binding sites gave equivocal results but suggested an increase in the high-affinity component. Analysis using the negative co-operativity binding model of De Meyts, Bianco and Roth [J. Biol. Chem. (1976) 251, 1877-1888] suggested that the affinity at both high and low occupancy was increased equally. The kinetics of association of insulin with the plasma-membrane receptor indicated that, although the net rate of association increased in the presence of chloroquine, this was due to a reduction in the dissociation rate rather than an increase in the association rate. This was confirmed by direct measurement of the rates of dissociation. Dissociation was found to be distinctly biphasic, with fast and slow components. Curve fitting suggested that the decrease in dissociation rate in the presence of chloroquine was not due to a decrease in either of the two dissociation rate constants, but rather to an increase in the amount of insulin dissociating by the slow component. It was also found that the increase in dissociation rate in the presence of excess insulin, ascribed to negative co-operativity, could be accounted for by an increase in the amount of insulin dissociating by the faster pathway, rather than by an increase in the dissociation rate constant. Thus chloroquine appears to have the opposite effect to excess insulin, and evidence was found for the induction of positive co-operativity in the insulin-receptor interaction at high chloroquine concentrations. Evidence was also found for the presence of low-affinity chloroquine binding sites with binding parameters similar to the concentration dependence of the chloroquine-induced augmentation of insulin binding. PMID- 7487934 TI - Distinct functions of the 90 kDa heat-shock protein (hsp90) in oestrogen and mineralocorticosteroid receptor activity: effects of hsp90 deletion mutants. AB - Recent studies have confirmed that the 90 kDa heat-shock protein (hsp90) interacts both in vitro and in vivo with steroid receptors, encouraging further detailed physicochemical and functional analysis of its chaperone role. Thus, to explore the relationship between hsp90 and receptors, the baculovirus system was used to overexpress the chick hsp90 alpha (chsp90) along with the chick oestradiol receptor (cER) or the human mineralocorticosteroid receptor (hMR). These receptors were able to form 9 S complexes with chsp90, demonstrating the association of the co-expressed recombinant proteins. Three mutants of chsp90 (delta A, delta B and delta Z) have been created by deletion of the A (residues 221-290) and B (530-581) regions, rich in charged amino acids, and the Z (392 419) region, a putative leucine zipper. After co-expression, anti-receptor antibodies immunoprecipitated the cER or hMR complexed with the wild-type chsp90, the delta B or the delta Z mutant, but not with the delta A chsp90, indicating that deletion of the A region of chsp90 leads to a lack of interaction with these receptors. The hormone binding capacity of the cER was unaffected after its co expression with each of the three mutants. In contrast, the hMR co-expressed with the delta B mutant failed to bind aldosterone, a finding confirmed in vivo by the absence of hormone-induced hMR nuclear translocation. Thus the B region is required for high-affinity ligand binding by the hMR. Our results suggest that the A region (but not the B or Z regions) is involved in binding of chsp90 to the cER and hMR, while the B region is essential for hormone binding by the hMR, consistent with a chaperone function for hsp90. PMID- 7487935 TI - Sequence and expression of the mRNA encoding HSP22, the mitochondrial small heat shock protein in pea leaves. AB - A 3 h treatment at 40 degrees C of pea (Pisum sativum var. Douce Provence) plants induces production and accumulation of a small heat-shock protein of 22 kDa apparent molecular mass, designated HSP22, in the matrix compartment of mitochondria [Lenne and Douce (1994) Plant Physiol. 105, 1255-1261]. We show here that the HSP22 precursor (i.e. the mature protein plus the transit peptide) has an apparent molecular mass of 26 kDa after in vitro translation of mRNA extracted from heat-stressed pea plants and immunodetection. We have isolated, cloned and sequenced the full-length cDNA encoding the precursor of the mitochondrial HSP22. An analysis of the amino acid sequence of the mitochondrial HSP22 reveals that this protein is a representative member of the low-molecular-mass heat shock protein (HSP) superfamily, exhibiting the specific consensus regions that are typical of the small HSPs. Most importantly, comparison of the mitochondrial HSP22 sequence with that of chloroplast small HSPs indicates that HSP22 does not contain the typical chloroplast consensus region III. We have also analysed the kinetics of HSP22 induction, and report results on the temporal expression of HSP22 at the transcriptional level. HSP22 mRNA was detected as soon as 10 min after the temperature was raised to a high temperature of 40 degrees C. Then the amount of HSP22 mRNA declined considerably even though pea plants were still submitted to the heat treatment. These results are discussed in light of the translation data previously published [Lenne and Douce (1994) Plant Physiol. 105, 1255-1261], particularly concerning the physiological behaviour of mitochondria when plants are heat-stressed. Furthermore, we have studied the dependence of HSP22 accumulation with temperature and demonstrate that the pea mitochondrial heat-shock response is only developed under extreme environmental growth conditions. PMID- 7487936 TI - Substitutions of aspartic acid for glycine-220 and of arginine for glycine-664 in the triple helix of the pro alpha 1(I) chain of type I procollagen produce lethal osteogenesis imperfecta and disrupt the ability of collagen fibrils to incorporate crystalline hydroxyapatite. AB - We identified two infants with lethal (type II) osteogenesis imperfecta (OI) who were heterozygous for mutations in the COL1A1 gene that resulted in substitutions of aspartic acid for glycine at position 220 and arginine for glycine at position 664 in the product of one COL1A1 allele in each individual. In normal age- and site-matched bone, approximately 70% (by number) of the collagen fibrils were encrusted with plate-like crystallites of hydroxyapatite. In contrast, approximately 5% (by number) of the collagen fibrils in the probands' bone contained crystallites. In contrast with normal bone, the c-axes of hydroxyapatite crystallites were sometimes poorly aligned with the long axis of fibrils obtained from OI bone. Chemical analysis showed that the OI samples contained normal amounts of calcium. The probands' bone samples contained type I collagen, overmodified type I collagen and elevated levels of type III and V collagens. On the basis of biochemical and morphological data, the fibrils in the OI samples were co-polymers of normal and mutant collagen. The results are consistent with a model of fibril mineralization in which the presence of abnormal type I collagen prevents normal collagen in the same fibril from incorporating hydroxyapatite crystallites. PMID- 7487937 TI - Biological fate of amino acid, peptide and protein hydroperoxides. AB - In the course of searching for a suitable marker for studying protein oxidation, we have successfully elucidated the structures of three valine hydroperoxides, i.e. beta-hydroperoxyvaline, (2S,3S)-gamma-hydroperoxyvaline and (2S,3R)-gamma hydroperoxyvaline, which are novel products of protein oxidation. The corresponding valine hydroxides were obtained by sodium borohydride reduction [Fu, Hick, Sheil and Dean (1995) Free Rad. Biol. Med. 19, 281-292]. We hypothesized that valine hydroxides might be the major biological degradation products of valine hydroperoxides and, as such, could be useful markers for the study of protein oxidation in vivo. The aim of this study was to investigate the fate of valine hydroperoxide in selected biological systems by the use of chemiluminescence detection of hydroperoxides and HPLC analysis of O phthaldialdehyde derivatives of amino acid residues. The degradation of hydroperoxides present on gamma-radiolysed solutions of valine, Pro-Val-Gly, or BSA occurred in the presence of: (1) transition metals (Fe2+, Fe3+, or Cu2+), (2) the detoxifying enzyme GSH peroxidase, (3) human plasma, and (4) J774 mouse monocyte macrophage cells. The major degradation product of valine hydroperoxide recovered in each case was found to be a valine hydroxide. These results suggest that valine hydroxide (derived from the hydroperoxide) may well be a useful in vivo marker for studying protein damage under oxidative stress. PMID- 7487938 TI - Permissive effect of ceramide on growth factor-induced cell proliferation. AB - Addition of bacterial sphingomyelinase to quiescent Swiss 3T3 cells effectively potentiated the platelet-derived growth factor (PDGF)-stimulated cell proliferation, though the enzyme by itself had little effect on the cell proliferation. Such potentiation of the cell growth could also be observed by the addition of ceramide, a product of the sphingomyelinase-catalysed reaction. In contrast, phosphocholine, another product of the enzyme reaction, had no synergistic effect on the action of PDGF. Treatment of the cells with sphingomyelinase or ceramide increased the cellular activity of mitogen-activated protein kinases (MAP kinases), which have been implicated in the regulation of cell proliferation. However, the synergistic effect of sphingomyelinase on the PDGF-induced cell growth could still be observed even when the cellular MAP kinase activity was fully activated by the growth factor alone. These results indicate that a ceramide-mediated cellular event(s) other than the MAP kinase activation is potentially involved in the regulation of cell growth. PMID- 7487939 TI - Human dipeptidyl peptidase IV gene promoter: tissue-specific regulation from a TATA-less GC-rich sequence characteristic of a housekeeping gene promoter. AB - The dipeptidyl peptidase IV gene encodes a plasma-membrane exopeptidase that is highly expressed in small intestine, lung and kidney. In order to better understand the mechanisms responsible for this tissue-specific expression we cloned, sequenced and functionally characterized the 5'-flanking region of the human dipeptidyl peptidase IV gene. The first 500 bases of the 5'-flanking sequence constituted an unmethylated CpG island, contained several Sp1-binding sites and lacked a consensus TATA box, all characteristics of gene promoters lacking tissue-specific expression. RNase-protection analysis using both small intestinal and Caco2 cell RNA indicated that the dipeptidyl peptidase IV transcript was initiated from no fewer than six major and 12 minor start sites. The 5'-flanking sequence also exhibited functional promoter activity in transient transfection experiments. Here, various lengths of the sequence were cloned upstream of a luciferase gene and introduced into cultured cells using lipofectin. A region located between bases -150 and -109 relative to the start of translation was found to be important for high-level promoter activity in both Caco2 and HepG2 cells. Moreover, Caco2 cells and HepG2 cells, which express high levels of dipeptidyl peptidase IV activity, exhibited much higher normalized luciferase activity after transfection than did 3T3, Jurkat or COS-7 cells, which have low enzyme levels. Sodium butyrate was found to increase both enzyme activity and normalized luciferase in HepG2 cells. Thus the dipeptidyl peptidase IV promoter possesses the ability to initiate transcription in a tissue-specific fashion in spite of having the sequence characteristics of a housekeeping gene promoter. PMID- 7487940 TI - Effects of C-1-substituted glucose analogue on the activation states of glycogen synthase and glycogen phosphorylase in rat hepatocytes. AB - A series of glucose-analogue inhibitors of glycogen phosphorylase b (GPb) has been designed, synthesized and investigated in crystallographic binding and kinetic studies. The aim is to produce a compound that may exert more effective control over glycogen metabolism than the parent glucose molecule and which could alleviate hyperglycaemia in Type-II diabetes. N-Acetyl-beta-D-glucopyranosylamine (1-GlcNAc) has a Ki for muscle GPb in crude extracts of 30 microM, 367-fold lower than that of beta-D-glucose [Board, Hadwen and Johnson (1995) Eur. J. Biochem. 228, 753-761]. In the current work, the effects of 1-GlcNAc on the activation states of GP and glycogen synthase (GS) in cell-free preparations and in isolated hepatocytes are reported. In gel-filtered extracts of liver, which lack ATP for kinase activity, 1-GlcNAc produced a rapid and time-dependent inactivation of GP with a subsequent activation of GS. Effects of 1-GlcNAc on both enzymes were stronger than those of glucose, with 0.8 mM 1-GlcNAc being equipotent with 50 mM glucose. At 1 mM, 1-GlcNAc enhanced the dephosphorylation of exogenous GPa by liver extracts (600%) and by muscle extracts (75%). This represents an approximately 500-fold improvement on glucose for the liver activity and 40-fold for the muscle activity. In whole hepatocytes, 1-GlcNAc showed an approximately 5 fold enhancement of glucose effects for GP inactivation but failed to elicit activation of GS. Glucose-induced activation of GS in whole hepatocytes was reversed by subsequent addition of 1-GlcNAc. However, when GS activation was achieved via the adenosine analogue and kinase inhibitor, 5'-iodotubercidin (ITU), subsequent addition of 1-GlcNAc allowed continued activation of GS. Phosphorylation of 1-GlcNAc in rat hepatocytes was established using radiolabelled material. The rate of phosphorylation was 1.60 nmol/min per 10(6) cells at 20 mM 1-GlcNAc but was reduced by the presence of 50 microM ITU (0.775 nmol/min per 10(6) cells). It is suggested that the phosphorylated derivative of 1-GlcNAc formed in hepatocytes is 1-GlcNAc 6-phosphate and that the presence of this species is responsible for the failure of 1-GlcNAc to activate GS. The relative importance of the reduction in concentration of GPa versus increased glucose 6-phosphate levels for activation of GS is discussed. PMID- 7487941 TI - Flexibility of zonation of fatty acid oxidation in rat liver. AB - Periportal and perivenous hepatocytes were isolated from rats subjected to different treatments that induce (starvation, cold exposure) or depress (refeeding after starvation) hepatic fatty acid oxidation. These experiments were designed to determine factors that may be involved in creating and maintaining the asymmetrical distribution of this metabolic pathway in the acinus of the liver. The uneven distribution of mitochondrial [14C]-palmitate oxidation within the acinus (i) was very flexible and changed markedly with the physiological status of the animal (periportal/perivenous ratio: 1.5, 2.0, 1.0 and 0.4 for fed, starved, refed and cold-exposed animals respectively), (ii) coincided with a similar zonation of carnitine palmitoyltransferase I activity in fed as well as in cold-exposed animals, (iii) was paralleled by a comparable zonation of mitochondrial 3-hydroxy-3-methyl-glutaryl-CoA synthase activity in starved animals, and (iv) was not determined by zonal differences in any of the following parameters: sensitivity of carnitine palmitoyltransferase I to malonyl-CoA, intracellular concentration of malonyl-CoA, fatty acid synthesizing capacity, acetyl-CoA carboxylase activity, fatty acid synthase activity or relative content of the two hepatic acetyl-CoA carboxylase isoforms. Unlike mitochondrial oxidation, peroxisomal [14C]palmitate oxidation was always zonated towards the perivenous zone of the liver irrespective of the physiological status of the animal. The data presented show that changes in the acinar distribution of mitochondrial long-chain fatty acid oxidation involve specific long-term mechanisms under different physiological conditions. PMID- 7487943 TI - An anti-neoplastic glycan isolated from Mycobacterium bovis (BCG vaccine). AB - Tice substrain BCG is used clinically as an immunotherapeutic agent against superficial bladder cancer. A boiling-water extract of this BCG showed anti tumour activity against a murine S180 sarcoma model and was fractionated into three fractions, A, B and C, by the use of Sephadex LH-20 chromatography. An anti tumour glucan, PS1A1, was isolated from fraction PS1A with Sephadex G-75. The molecular mass of PS1A1 was between 65 and 87 kDa by Sephadex G-100 chromatography. The structure of PS1A1 was investigated by one- and two dimensional NMR spectroscopy and methylation analysis and was demonstrated to be primarily 1-->6-alpha-linked glucose units. We postulate that the repeating unit is: [Formula: see text] PMID- 7487942 TI - Heterologous expression of rat liver microsomal glutathione transferase in simian COS cells and Escherichia coli. AB - The cDNA coding for rat liver microsomal glutathione transferase was subcloned into the mammalian expression vector pCMV-5 and the construct was transfected into, and transiently expressed in, simian COS cells. This resulted in high expression (0.7% of the microsomal protein). The activity towards 1-chloro-2,4 dinitrobenzene in microsomes was 15-30 nmol/min per mg, which increased upon N ethylmaleimide treatment to 60-200 nmol/min per mg. Control and antisense-vector treated cells displayed very low activity (3-6 nmol/min per mg). A DNA fragment coding for rat microsomal glutathione transferase was generated by PCR, cloned into the bacterial expression vector pSP19T7LT and transformed into Escherichia coli strain BL21 (DE3) (which contained the plasmid pLys SL). Isopropyl beta-D thiogalactopyranoside (IPTG; 1 mM) induced the expression of significant amounts of enzymically active protein (4 mg/l of culture as measured by Western blots). The recombinant protein was purified and characterized and found to be indistinguishable from the rat liver enzyme with regard to enzymic activity, molecular mass and N-terminal amino acid sequence. Human liver cDNA was used to obtain the coding region of human microsomal glutathione transferase by PCR. This PCR product was cloned into pSP19T7LT, which, upon induction with IPTG, yielded significant amounts (9 mg/l of culture) of active enzyme in BL21 (DE3) cells. Thus, for the first time, it is now possible to express both human and rat microsomal glutathione transferase in an enzymically active form in Escherichia coli. PMID- 7487944 TI - Two new sphingomyelin analogues inhibit phosphatidylcholine biosynthesis by decreasing membrane-bound CTP: phosphocholine cytidylyltransferase levels in HaCaT cells. AB - The effects of two newly synthesized sphingomyelin analogues on phosphatidylcholine biosynthesis were investigated in the immortalized human keratinocyte cell line HaCaT. N-Acetyl-erythro-sphingosine-1-phosphocholine (AcSM) and N-octanoyl-erythro-sphingosine-1-phosphocholine (OcSM) inhibited the incorporation of choline into phosphatidylcholine with half-inhibitory concentrations (IC50) of 6 micrograms/ml and 10 micrograms/ml respectively. Further experiments revealed that AcSM and OcSM interfered with the translocation of the rate-limiting enzyme of phosphatidylcholine biosynthesis, CTP:phosphocholine cytidylyltransferase (EC 2.7.7.15), in HaCaT cells and inhibited cytidylyltransferase activity in vitro. Despite the fact that OcSM was a potent inhibitor of cytidylyltransferase in vitro, its effects on phosphatidylcholine biosynthesis and translocation of cytidylyltransferase in HaCaT cells were less pronounced as compared with AcSM. Finally, we showed that the comparatively strong effects of AcSM in cell culture experiments were due to the uptake of large amounts of this sphingomyelin analogue into the cells. The results presented demonstrate that the activity of cytidylyltransferase may be negatively regulated by a high ratio of choline head group-containing sphingolipids. PMID- 7487945 TI - Proteoglycans in polarized epithelial Madin-Darby canine kidney cells. AB - Madin-Darby canine kidney (MDCK) cells were cultured on polycarbonate filters to study the synthesis and sorting of proteoglycans in polarized epithelial cells. Two strains of MDCK cells were used. MDCK I cells resemble distal tubule epithelial cells, and MDCK II cells share some characteristics with proximal tubule cells. Both strains were grown to confluency and labelled with [35S]sulphate for 24 h. The apical and basolateral media and the cell fractions were harvested and analysed by DEAE ion-exchange chromatography. A large portion of the [35S]sulphate-labelled macromolecules bound strongly to the ion-exchange columns, and could be eluted in three distinct peaks. The latest eluting peak was demonstrated to contain almost exclusively chondroitin sulphate, whereas peak 2 contained mostly heparan sulphate, demonstrated by using chondroitinase ABC and nitrous acid (pH 1.5) respectively to depolymerize the [35S]glycosaminoglycan chains. Peak 1 contained negligible amounts of proteoglycans. Large differences could be observed in proteoglycan sorting in MDCK I and II cells. Strain I secreted approx. 67% of the proteoglycans to the apical side and 17% to the basolateral side. The cell fraction contained 17% of the proteoglycans after 24 h of labelling. In contrast, 19% of the proteoglycans were sorted to the apical side of MDCK II cells and 61% to the basolateral side, whereas the cell fraction contained 20%. Furthermore, the level of [35S]proteoglycan biosynthesis (apical and basolateral media and cell fraction total) was higher in MDCK I cells than in strain II. Based on the amount of material degraded by chondroitinase ABC and nitrous acid respectively, and the total amounts of [35S]proteoglycans recovered from the cells, it was calculated that the MDCK I strain synthesized approx. 56% chondroitin sulphate and 44% heparan sulphate. In contrast, the MDCK II strain synthesized 69% heparan sulphate and 31% chondroitin sulphate. To further identify the [35S]proteoglycans synthesized by MDCK I and II cells, antibodies against perlecan, versican and syndecan were used. The antibody against mouse syndecan did not cross-react with any of the proteoglycans produced in MDCK I or II cells. Both MDCK I and II cells expressed perlecan; 57-61% could be recovered from the basolateral fractions and 18-34% from the apical medium. Versican was also found in both MDCK I and II cells. Compared with perlecan, a larger percentage of versican (43-53%) was found in the cell fractions. PMID- 7487946 TI - The content of glutathione and glutathione S-transferases and the glutathione peroxidase activity in rat liver nuclei determined by a non-aqueous technique of cell fractionation. AB - Hepatocellular nuclei require glutathione, glutathione S-transferases (GSTs) and Se-dependent glutathione peroxidase (GPx) for intranuclear protection against damage from electrophiles or products of active oxygen. Data so far available from the literature on nuclei isolated in aqueous systems range from glutathione, GSTs and GPx either being absent altogether to being present in quantities in excess of those in the cytoplasm. This paper describes a small-scale preparation of a nuclear fraction from rat liver by a non-aqueous technique, designed to retain nuclear water-soluble molecules in situ, since low-molecular-mass compounds can diffuse freely into other compartments during aqueous separation. This non-aqueous procedure shows the nucleus to contain glutathione at 8.4 mM and soluble GSTs at 38 micrograms/mg of protein, the enrichment over the homogenate being 1.2-1.4-fold. Se-dependent GPx activity was also present in the nucleus (56 m-units/mg), although with slightly lower activity than in the homogenate (0.7 fold). PMID- 7487947 TI - Molecular cloning and characterization of a neurotoxic phospholipase A2 from the venom of Taiwan habu (Trimeresurus mucrosquamatus). AB - Using gel-filtration chromatography and reverse-phase (RP) HPLC we have purified a presynaptic neurotoxin (designated as trimucrotoxin) from the crude venom of Taiwan habu (Trimeresurus mucrosquamatus). Its complete primary structure was solved by an automated N-terminal sequencing and cDNA sequencing method. The enzyme inhibited the twitch of the chick biventer cervicis muscle at 0.1-1 micrograms/ml and showed lethality in mice (LD50 = 1.2 micrograms/g, when given intravenously). Trimucrotoxin exists mainly as a homodimer of 14 kDa subunits as shown by a gel-filtration experiment, and dissociates into monomers during SDS/PAGE in the absence of Ca2+. However, most of trimucrotoxin migrated as slowly as a trimer during nondenaturing SDS/PAGE in the presence of Ca2+ or Sr2+. Its amino acid sequence identity to crotoxin B and agkistrodotoxin is about 75%, and its cDNA sequence is 82% identical to that of crotoxin B. Rabbit antiserum against trimucrotoxin also cross-reacted with the other crotalid neurotoxic phospholipases A2. Furthermore, the purified acidic subunit of crotoxin potentiated the neurotoxicity of trimucrotoxin. A comparison of the sequences of these crotalid neurotoxins revealed some common features of the possible neurotoxic sites, including residues 6, 11, 76-81 and 119-125. PMID- 7487948 TI - Identification of direct-repeat-binding protein 1 (DRP-1), a DNA-binding protein that binds specifically to the 'malic' enzyme gene promoter direct repeat element. AB - The 'malic' enzyme (ME) gene promoter contains three main regulatory regions. One of these, the direct repeat element (DRE), contains tandem degenerate Sp1-binding sites separated by a 3 bp intervening sequence. We now show that a previously unreported 95 kDa protein, which we have designated DRP-1, binds strongly to the DRE region in a highly specific manner. Western-blot analysis confirms that this protein is not Sp1, which has been shown to bind to similar degenerate sites. Competitive binding assays using purified DRP-1 further reveal that neither non specific nor Sp1-consensus-site-containing oligonucleotides can displace those complexes formed between DRP-1 and the DRE sequence, thus confirming sequence specific binding by this protein. SDS/PAGE analysis of DRE-protein complexes isolated by direct excision and transplantation from retardation gels confirms the presence of the 95 kDa protein and, in addition, suggests that more than one binding site exists for this protein within the DRE. This is in accord with the repeated nature of the DRE DNA sequence which contains two CACC box motifs. PMID- 7487949 TI - Urea-induced inactivation and denaturation of clostridial glutamate dehydrogenase: the absence of stable dimeric or trimeric intermediates. AB - Urea-induced effects in clostridial glutamate dehydrogenase (GDH, EC 1.4.1.2) were studied by spectrophotometry, circular dichroism, FPLC, affinity chromatography and PAGE. Denaturation of enzyme occurred over a narrow range of urea concentrations (2.5-3.5 M), accompanied by inactivation of enzyme with a similar rate constant. The contribution of instantaneous inhibition by urea was also ascertained. FPLC studies of urea-treated GDH gave no evidence for dissociated oligomeric fragments of the hexamer in the presence of subdenaturing concentrations of urea. Likewise a mixture of fully 5,5'-dithiobis-(2 nitrobenzoic acid)-modified GDH hexamers and unmodified enzyme in 2 M urea failed to give rise to hybrid molecules. Exposure of unmodified GDH to high concentrations of urea led to the dissociation of hexamers to denatured monomers followed by association to form non-specific high-M(r) aggregates. This conclusion was confirmed by native gradient PAGE experiments. Various specific ligands stabilized the enzyme against urea-induced inactivation, succinate and 2 oxoglutarate being particularly effective. This protection of the native state was enhanced in ternary complexes, and the complex most resistant to urea-induced inactivation was the productive ternary complex GDH-NADH-2-oxoglutarate. Native gradient PAGE experiments indicate that these protecting ligands preserve the native hexameric structure of GDH. PMID- 7487950 TI - The selective mobilization of fatty acids is not based on their positional distribution in white-fat-cell triacylglycerols. AB - Fatty acids have been shown to be selectively mobilized from rat white fat-cells, whatever the dietary manipulations. For convenience, fatty acids have been classified as being highly, weakly and moderately mobilizable. The aim of this study was to examine whether the selective mobilization of fatty acids can be explained, even partly, by their positional distribution in adipose-tissue triacylglycerols (TAG) via the known specificity of hormone-sensitive lipase for the sn-1 and sn-3 positions. Adipose tissue was dietarily manipulated in order to obtain a wide spectrum of fatty acids, including large amounts of either very long-chain polyunsaturated fatty acids (VLC-PUFA) or very-long-chain monounsaturated fatty acids (VLC-MUFA). The determination of fatty acid distribution in adipose tissue TAG was based on random formation of 1,2-diacyl rac-glycerols by Grignard degradation, followed by synthesis of phosphatidic acids and hydrolysis in the sn-2 position by phospholipase A2. Regardless of the fatty acid composition and location of fat depots, highly (e.g. 18:4n-3 and some of the VLC-PUFA) and weakly (e.g. VLC-MUFA) mobilizable fatty acids were located mainly in the outer (sn-1 and sn-3) positions of the glycerol moiety (79.5% and 92.5% on average, respectively). Other fatty acids, which are rather moderately mobilizable, were more randomly distributed. We conclude that the selective mobilization of white-fat-cell fatty acids is not based on their positional distribution in TAG. PMID- 7487951 TI - A model for the initial phase of cell/surface interactions based on ligand binding phenomena. AB - The interaction between mammalian cells and solid surfaces plays an important role in a number of biological phenomena. Of particular clinical importance is the migration of cells suspended in blood to the wall of a blood vessel in the event of tissue damage. While the resultant inflammation often represents a desirable response to an external challenge, responses of this type can also lead to adverse consequences. Although the cell migration phenomenon is well known, a plausible mechanism for controlling the critical 'rolling' stages of adhesion has yet to be proposed. In this report we suggest how a simple consideration of ligand/receptor binding interactions can be used to explain a switch between a situation where a cell population is almost entirely in free suspension, to one where a significant fraction is attached to the solid surface. PMID- 7487952 TI - The novel cyclic dinucleotide 3'-5' cyclic diguanylic acid binds to p21ras and enhances DNA synthesis but not cell replication in the Molt 4 cell line. AB - 1. The effect of the novel, naturally occurring nucleotide 3'-5' cyclic diguanylic acid (c-di-GMP) on the lymphoblastoid Molt 4 cell line was studied. When exposed to this guanine nucleotide. Molt 4 cells exhibited a marked increase in [3H]thymidine incorporation, up to 200-fold at 50 microM c-di-GMP. Correspondingly, the DNA content of the treated cells was 9-fold higher than untreated cells. Stimulation of [3H]thymidine incorporation into the cells was time- and concentration-dependent. This effect was specific and was not observed with GMP or cyclic GMP, nor with the unhydrolysable GTP analogues, guanosine 5' [gamma-thio]triphosphate and guanosine 5'-[beta gamma-imido]-triphosphate. C-di GMP entrance into the cells was experimentally verified and occurred without using any means of cell permeabilization. SDS/PAGE analysis of cells exposed to [32P]c-di-GMP, followed by autoradiography, revealed the labelling of three low molecular-mass proteins at 18-27 kDa. The labelling is highly specific to c-di GMP and its extent was not affected by other guanine nucleotides. 2. One of the c di-GMP-binding proteins was found to be the p21ras protein, by immunoprecipitation with the anti-Ras monoclonal antibody Y13-259. The effects described appear to be unique for c-di-GMP and, taken together, raise the possibility that an irreversible binding of this guanine nucleotide to the growth promoting p21ras protein results in a fixed active conformation of this protein affecting DNA synthesis. Strikingly, although at 48 h of growth markedly high DNA levels were found in Molt 4 cells treated with c-di-GMP, this guanine nucleotide had no effect on cell replication during this period. Thus Molt 4 cells exposed to c-di-GMP enter the S phase uncoordinated with their overall replication rate. PMID- 7487954 TI - Abnormal type III collagen produced by an exon-17-skipping mutation of the COL3A1 gene in Ehlers-Danlos syndrome type IV is not incorporated into the extracellular matrix. AB - A novel heterozygous mutation of the COL3A1 gene that encodes the alpha 1(III) chains of type III collagen was identified in a family with the acrogeric form of Ehlers-Danlos syndrome type IV (EDS-IV). Cultured dermal fibroblasts produced normal and shortened alpha 1(III) chains. The triple helix of the latter chain was shortened owing to a 33 amino acid deletion of Gly-184 to Pro-216. The corresponding region of cDNA lacked 99 base pairs from nucleotides 1051 to 1149. The deletions corresponded exactly to the normal sequence encoded by exon 17 of the COL3A1 gene. The proband was heterozygous for a T to G transversion at position +2 of intron 17, which resulted in skipping of exon 17. The splicing defect was not corrected by growing the fibroblasts at 33 degrees C and no other splicing variants were identified at 33 or 37 degrees C. The affected brother had the same mutation but his unaffected mother did not. Heterotrimeric type III collagen molecules containing normal and mutant chains were retained within the cell. The mutant homotrimeric molecules were modified and secreted normally and were thermally stable. These normal characteristics of the mutant homotrimers suggested that the loss of ten Gly-Xaa-Yaa triplets (where Gly-Xaa-Yaa is a repetitive amino acid triplet structure in which Xaa and Yaa are other amino acids, proline and hydroxyproline being more common in the Yaa position) did not adversely affect the formation and stability of the triple helix or the structural requirements for secretion. However, the mutant homotrimers were not incorporated into the extracellular matrix of an in vitro model of EDS-IV dermis. The EDS-IV phenotype in this family was probably due to a deficiency in the amount of normal type III collagen available for formation of the heterotypic collagen fibrils of the extracellular matrix. Intracellular and extracellular quality-control mechanisms prevented the incorporation of heterotrimeric and homotrimeric mutant type III collagen molecules into the cross-linked extracellular matrix. PMID- 7487953 TI - Expression analysis of the individual bovine beta-, alpha s2- and kappa-casein genes in transgenic mice. AB - To identify cis-acting regulatory elements involved in the regulation of expression of the casein genes, the bovine beta-, alpha s2- and kappa-casein genes were isolated from cosmid libraries and introduced into the murine germline. Bovine casein expression was analysed at the RNA and protein level. The bovine beta-casein gene, including 16 kb of 5'- and 8 kb of 3'-flanking region, appeared to be expressed in all 12 transgenic mouse lines analysed. In 50% of these lines expression levels in milk exceeded 1 mg/ml. Three lines displayed expression levels comparable with or well above (20 mg/ml) the beta-casein levels in bovine milk. Transgene expression was restricted to the mammary gland. Strong induction of expression occurred at parturition and thus resembled the bovine rather than the murine pattern. In spite of this high-level tissue-specific and developmentally regulated expression, beta-casein expression levels were integration-site-dependent, suggesting that not all elements involved in regulation of expression were included in this beta-casein clone. Neither the bovine alpha s2- nor the kappa-casein gene, including 8 kb and 5 kb of 5'- and 1.5 kb and 19 kb of 3'-flanking sequences respectively, were properly expressed in transgenic mice. However, they were transcribed in stably transfected mouse mammary epithelial cells. This indicates that regulatory elements required for high-level, mammary gland-specific expression are not present in the alpha s2- and kappa-casein clones used in this study and are probably located elsewhere in the casein gene locus. PMID- 7487955 TI - Early signalling mechanism in colonic epithelial cell response to gastrin. AB - The hormone gastrin exerts a growth-promoting effect on gastrointestinal cells. The molecular mechanisms by which colonic epithelial cells respond to gastrin are still poorly understood. In this study, we demonstrate a novel feature of the action of gastrin on normal colonic cells, namely the rapid phosphorylation on tyrosine of phospholipase C gamma 1 (PLC gamma 1). Tyrosine phosphorylation of PLC gamma 1, elicited by gastrin, was transient, concentration-dependent, and was abrogated by pretreating the colonic cells with the gastrin-receptor antagonist proglumide, the tyrosine kinase inhibitor genistein, and by removal of the tyrosine phosphatase inhibitor orthovanadate from the isolation buffer. Tyrosine phosphorylation of PLC gamma 1 correlated with the time- and concentration dependent decrease in the mass of membrane phosphatidylinositol 4,5-bisphosphate (PIP2) and the increase in the epithelial concentration of inositol 1,4,5 trisphosphate (IP3). Likewise, the stimulated increase in IP3 was also prevented by proglumide and genistein. Gastrin induced a definite but transient increase in the intracellular concentration of free Ca2+ [Ca2+]i, and increased membrane translocation of immunoreactive alpha- and beta-protein kinase C. The data thus indicate that gastrin elicits at least one signalling cascade, through rapid tyrosine phosphorylation of PLC gamma 1, leading to the activation of a PIP2 specific PLC pathway. PMID- 7487957 TI - Involvement of oligomerization, N-glycosylation and sialylation in the clearance of cholinesterases from the circulation. AB - The possible role of post-translational modifications such as subunit oligomerization, protein glycosylation and oligosaccharide processing on the circulatory life-time of proteins was studied using recombinant human acetylcholinesterase (rHuAChE). Different preparations of rHuAChE containing various amounts of tetramers, dimers and monomers are cleared at similar rates from the circulation, suggesting that oligomerization does not play an important role in determining the rate of clearance. An engineered rHuAChE mutant containing only one N-glycosylation site was cleared from the circulation more rapidly than the wild-type triglycosylated enzyme. On the other hand, hyperglycosylated mutants containing either four or five occupied N-glycosylation sites, analagous to those present on the slowly cleared fetal bovine serum acetylcholinesterase (FBS-AChE), were also cleared more rapidly from the bloodstream than the wild-type species. Furthermore, the two different tetraglycosylated mutants were cleared at different rates while the pentaglycosylated mutant exhibited the most rapid clearance profile. These results imply that though the number of N-glycosylation sites plays a role in the circulatory life-time of the enzyme, the number of N-glycan units in itself does not determine the rate of clearance. When saturating amounts of asialofetuin were administered together with rHuAChE, the circulatory half-life of the enzyme was dramatically increased (from 80 min to 19 h) and was found to be similar to that displayed by plasma-derived cholinesterases while desialylation of these enzymes caused a sharp decrease in the circulatory half-life to approximately 3-5 min. Determination of the average number of sialic acid residues per enzyme subunit of the five different N-glycosylation species generated, revealed that the rate of clearance is not a function of the absolute number of appended sialic acid moieties but rather of the number of unoccupied sialic acid attachment sites per enzyme molecule. Specifically, we demonstrate an inverse-linear relationship between the number of vacant sialic acid attachment sites and the values of the enzyme residence time within the bloodstream. PMID- 7487958 TI - Effect of irreversibility on the thermodynamic characterization of the thermal denaturation of Aspergillus saitoi acid proteinase. AB - The thermal denaturation of the acid proteinase from Aspergillus saitoi was studied by CD and differential scanning calorimetry (DSC). This process seemed to be completely irreversible, as protein samples that were heated to temperatures at which the transition had been completed and then cooled at 25 degrees C did not show any reversal of the change in the CD signal. Similar results were obtained with DSC. Nevertheless, we were able to detect the presence of reversibly unfolded species in experiments in which the enzyme solution was heated to a temperature within the transition region, followed by rapid cooling at 25 degrees C. Accordingly, the denaturation of behaviour of the acid proteinase seems to be consistent with the existence of one (or more) reversible unfolding transition followed by an irreversible step. The van't Hoff enthalpy, delta HvH, which corresponds to the reversible transition was calculated from extrapolation to infinite heating rate as 310 kJ.mol-1. This parameter was also determined from direct estimation of the equilibrium constant at several temperatures (delta HvH = 176 kJ.mol-1). Comparison of the average delta HvH with the calorimetric enthalpy (delta Hcal. = 770 kJ.mol-1) gave a value of 3.2 for the delta Hcal./delta HvH ratio, indicating that the molecular structure of the enzyme is probably formed by three or four cooperative regions, a number similar to that of the acid proteinase, pepsin. It should be noted that a completely different conclusion would be obtained from a straightforward analysis of the calorimetric curves, disregarding the effect of irreversibility on the denaturation process. PMID- 7487960 TI - Combination plots as graphical tools in the study of enzyme inhibition. AB - Although statistical regression has become the method of choice in the analysis of enzyme kinetics, graphical methods continue to be useful on account of their illustrative capabilities. It is pointed out in this paper that enzyme inhibition data may be presented more efficiently as a single linear plot than the traditional way as a family of lines. This approach has been taken previously by Hunter and Downs [Hunter and Downs (1945) J. Biol. Chem. 157, 427-446] but has remained neglected. A new version of this type of plot (combination plot) has been devised which is linear for competitive, non-competitive, uncompetitive and linear mixed inhibition and has a characteristic appearance for each type of inhibition behaviour. The slopes and intercepts not only indicate directly the dissociation constant but also provide quantitative criteria for the nature of inhibition. This plot should serve as a useful graphical tool in enzyme research as well as in biochemical education. PMID- 7487956 TI - Mass spectrometry and amino acid sequencing of two cadmium-binding metallothionein isoforms from the terrestrial gastropod Arianta arbustorum. AB - 1. Two cadmium-binding metallothionein (Mt) isoforms, called Mta and Mtb, were isolated from terrestrial snails (Arianta arbustorum), using various chromatographic techniques, such as gel-permeation chromatography and reversed phase HPLC. The purified proteins were S-methylated and cleaved by means of different enzymes (trypsin, endoproteinase Glu-C, and endoproteinase Asp-N). Amino acid sequences were determined by automated Edman degradation and collision induced dissociation (CID) tandem MS. According to their primary structures, both isoforms should be attributed to class-I Mts. 2. The two forms are structurally identical, differing only by one amino acid exchange in position 60 of the peptide chain. Both isoproteins consist of 66 amino acids, 18 of which are cysteine residues. Most of the cysteine residues are arranged in seven Cys-Xaa Cys motifs. Mta and Mtb possess an N-terminal acetylated-serine residue and contain a short N-terminal motif which shows a high degree of similarity with the N-termini of histones H4 and H2A. 3. A comparison of Mta and Mtb with other invertebrate Mts shows a very high degree of sequence similarity with a cadmium binding Mt from Helix pomatia, a species that is closely related to Arianta arbustorum. Moreover, Mta and Mtb, as expected, also exhibit structural similarities with Mts from other molluscan species, such as mussels and oysters. It is suggested that Mta and Mtb represent two allelic isoforms, reflecting the genetic polymorphism of Mt in Arianta arbustorum. PMID- 7487959 TI - Relationship between the microsomal epoxide hydrolase and the hepatocellular transport of bile acids and xenobiotics. AB - Recently two different bile-acid carriers for the hepatocellular sodium-dependent uptake of taurocholate have been described. The first transport system was isolated and characterized by functional expression cloning in Xenopus laevis oocytes. The corresponding cDNA clone, named Ntcp for Na+/taurocholate co transporting polypeptide, codes for a protein of 362 amino acids and shows no similarity to previously known sequences. The transport function of this carrier system is well documented by expression in Xenopus laevis oocytes and by transient and stably transfected cell lines. In addition, several lines of evidence implied that the well-known xenobiotic-metabolizing enzyme microsomal epoxide hydrolase (mEH, EC 3.3.2.3) is also able to mediate sinusoidal uptake of taurocholate. Furthermore, it was claimed that the same enzyme also mediates the uptake of the conjugated bile acid into the smooth endoplasmic reticulum (ER). No direct proof of the transport function of mEH by its heterologous expression has yet been published. In the present work we used a stable transfected cell line that expressed high levels of heterologous mEH for uptake studies of various bile acids and the loop diuretic bumetanide. The uptake of the conjugated bile acid taurocholate, of the non-conjugated bile acid cholate and of the organic anion bumetanide was measured in the transfected as well as in the non-transfected parental cell line. These organic anions represent the main substrates of the known transport systems for organic anions in the rat liver. The results show that the microsomal epoxide hydrolase is unable to transport taurocholate, cholate or bumetanide. Furthermore, Western-blot analysis revealed the expression of mEH in hepatoma tumor cell lines, which show no transport activity for these organic anions. These results show that it is unlikely that mEH can mediate the transport of these substrates. PMID- 7487963 TI - Tag team specification of a neural precursor in the Drosophila embryonic central nervous system. AB - The development of vertebrate and invertebrate nervous systems requires the production of thousands to millions of uniquely specified neurons from progenitor neural stem cells. A central question focuses on the elucidation of the developmental mechanisms that function within neural stem cell lineages to impart unique identities to neurons. A recent report details the roles that two genes, pdm-1 and pdm-2, play within an identified neural stem cell lineage in the Drosophila embryonic central nervous system. The results show that pdm-1 and pdm 2 are coexpressed in an identified neural precursor and function redundantly to specify the fate of this cell. As such this report offers an initial view of the genetic programs that create neural diversity. PMID- 7487962 TI - Prolonged elevation of intracellular cyclic AMP levels in U937 cells increases the number of receptors for and the responses to formylmethionyl leucylphenylalanine, independently of the differentiation process. AB - The effects of elevated levels of cyclic AMP induced by cholera toxin (CTx) were investigated on the differentiated promyelomonocytic cell line U937. After CTx treatment, the initial inhibition of the oxidative burst induced by N formylmethionyl-leucyl-phenylalanine (FMLP) was followed by a progressive increase over 20 h, resulting in 4-6-fold potentiation of the initial burst. Various cyclic-AMP-elevating agents produced similar potentiation of the FMLP- or C5a-induced oxidative burst, but the phorbol 12-myristate 13-acetate-induced oxidative burst was not affected by CTx pretreatment of cells. Furthermore, the increase in arachidonate release and intracellular Ca2+ triggered by FMLP were amplified after CTx treatment. ADP-ribosylation of Gi alpha subunits catalysed by pertussis toxin was slightly increased after CTx treatment, despite similar immunoreactivity of the alpha subunit of Gi2. FMLP binding sites present in CTx treated membranes were 3-6 times more abundant than in control membranes. Expression of mRNAs encoding the FMLP receptor and one of its related receptors were enhanced after CTx treatment of both undifferentiated and undifferentiated U937 cells. In parallel, after undifferentiated cells were treated with CTx, they were able to increase intracellular Ca2+, but not the oxidative burst, in response to FMLP. These data demonstrate that CTx, by increasing cyclic AMP, enhances the expression of chemotactic receptors independently of U937 cell differentiation. PMID- 7487961 TI - The photoactivatable NAD+ analogue [32P]2-azido-NAD+ defines intra- and inter molecular interactions of the C-terminal domain of the G-protein G alpha t. AB - Recently, we reported the synthesis and use of [32P]2-azido-NAD+ as a probe to study the structural organization of G-proteins. Pertussis toxin was used to 'tether' [32P]2-azido-ADP-ribose of [32P]2-azido-NAD+ to Cys347 of the alpha subunit of the G-protein Gt. Light activation of the azide moiety covalently cross-linked the domain containing Cys347 at the C-terminus of alpha t with neighbouring intra- and inter-molecular domains of holo-transducin. The radiolabel from [32P]2-azido-ADP-ribose was then transferred to the 'acceptor' domain by cleaving the thioglycosidic bond between Cys347 and [32P]2-azido-ADP- ribose with mercuric acetate. ADP-ribosylation followed by photocross-linking of holo-transducin indicated intramolecular interactions of the C-terminal domain with other alpha t domains and intermolecular interactions with holotransducin alpha and gamma subunits. The radiolabelled peptides, which were radiolabelled because of the transfer of the photoactive moiety, were identified by utilizing 2 (2'-nitrophenylsulphenyl)-3-methyl-3'- bromoindolenine ('BNPS-skatole') and CNBr. The results indicate that the C-terminus of alpha t interacts with both N terminal and C-terminal domains within the alpha t molecular. Mapping the interacting sites between cross-linked alpha dimers and alpha trimers indicates that the C-terminal domain of alpha t is involved in the formation of alpha t homopolymers in solution. In addition, our studies place the beta gamma subunit in close proximity to Cys347 of alpha t, as indicated by the transfer of [32P]2 azido-ADP-ribose from Cys347 to the gamma subunit, which was further localized to the C-terminal half of gamma t. The studies presented here identify the C terminal intra- and inter-molecular interactions of the alpha subunit of holo transducin. PMID- 7487964 TI - Reelin: a novel extracellular matrix protein involved in brain lamination. AB - Normal development of the nervous system is achieved through an elaborate program of guided neuronal migration and axonal growth. In the last few years, a flood of research has dissected the molecular bases of these phenomena, and several cell surface and extracellular matrix molecules, which are implicated in neuronal and axonal targeting processes, have been recognized. Taking this knowledge a step further, a recent paper by Tom Curran's group reports the molecular cloning of the gene deleted in the autosomal recessive mouse mutation reeler, affecting cortical neuronal migration. This gene encodes reelin, a novel extracellular matrix protein. PMID- 7487965 TI - Towards unravelling the Igf2/H19 imprinted domain. AB - Genomic imprinting is an epigenetic marking process that confers parent-of-origin dependent expression on certain genes. These imprinted genes are sometimes found in clusters, suggesting a possible involvement of higher order regulatory elements controlling expression and imprinting of genes organised in such clusters. In the distal chromosome 7 there are at least four imprinted genes: Mash2, Ins2, Igf2 and H19. Recent evidence suggests that imprinting and expression of at least Igf2 and H19 may be mechanistically linked. PMID- 7487967 TI - The specificity of homeotic gene function. AB - How transcription factors achieve their in vivo specificities is a fundamental question in biology. For the Homeotic Complex (HOM/Hox) family of homeoproteins, specificity in vivo is likely to be in part determined by subtle differences in the DNA binding properties inherent in these proteins. Some of these differences in DNA binding are due to sequence differences in the N-terminal arms of HOM/Hox homeodomains. Evidence also exists to suggest that cofactors can modify HOM/Hox function by cooperative DNA binding interactions. The Drosophila homeoprotein extradenticle (exd) is likely to be one such cofactor. In HOM/Hox proteins, both the conserved 'YPWM' peptide motif and the homeodomain are important for interacting with exd. Although exd provides part of the answer as to how specificity is achieved, there may be additional cofactors and mechanisms that have yet to be identified. PMID- 7487966 TI - Epidermal growth factor receptor function in early mammalian development. AB - We review here the data indicating a role for epidermal growth factor receptor (EGF receptor) signalling in early mouse development. Embryonic development of the metazoan embryo generally begins with the formation of a cystic structure and epithelial layers that subsequently form anlagen of the definitive body parts and organs. For the mammalian embryo, this cystic structure is a blastocyst whose wall consists of trophectoderm, the first epithelium to develop during mammalian embryogenesis. The onset of expression and function of EGF receptors is coincident with the onset of trophectoderm development. Modulating EGF receptor expression and function modulates trophectoderm differentiation, leading to the hypothesis that functional EGF receptors participate in the induction of trophectoderm development and perhaps of other embryonic epithelial derivatives such as nervous tissues. PMID- 7487968 TI - The complexity in regulating the expression of tenascins. AB - The tenascins are a growing family of extracellular matrix proteins of typical multidomain structure. The prototype to be discovered was tenascin-C. It shows a highly regulated expression pattern during embryonic development and is often transiently associated with morphogenetic tissue interactions during organogenesis. In the adult organism reexpression of tenascin-C occurs in tumors and many other pathological conditions. Tenascin-C expression can be regulated by many different growth factors and hormones. Furthermore, mechanical strain exerted by fibroblasts seems to induce the expression of tenascin-C. This could represent a mechanism of translating mechanical forces into protein patterns, a step of potential relevance in the organization of embryogenesis. Tenascin-C as well as tenascin-R are believed to counteract the cell adhesion and spreading activity of fibronectin, thereby facilitating cell movement. PMID- 7487969 TI - A unique role for enhancers is revealed during early mouse development. AB - Transcription and replication of genes in mammalian cells always requires a promoter or replication origin, respectively, but the ability of enhancers to stimulate these regulatory elements and the interactions that mediate this stimulation are developmentally acquired. The primary function of enhancers is to prevent repression, which appears to result from particular components of chromatin structure. Factors responsible for this repression are present in the maternal nucleus of oocytes and its descendant, the maternal pronucleus of mouse 1-cell embryos and in mouse 2-cell embryos, but are absent in the paternal pronucleus. Thus, enhancers are not needed to achieve efficient transcription and replication in paternal pronuclei. However, enhancers, even in the presence of their specific activation protein, are inactive prior to formation of a 2-cell embryo, suggesting that a coactivator essential for enhancer function is not available until zygotic gene expression begins. Furthermore, enhancer stimulation of transcription appears to be mediated through a promoter transcription factor, but this interaction can change as cells undergo differentiation, switching from a TATA-box independent to a TATA-box dependent mode. PMID- 7487970 TI - Dual mechanisms of lymphocyte-mediated cytotoxicity serve to control and deliver the immune response. AB - Cytotoxic lymphocytes play a central role in immune inflammatory responses against tumour cells, viruses and cells transplanted or infected with intracellular bacteria. The pivotal importance of lymphocytes in each of these immune responses has justified our continued interest in their cytotoxic function. Recent studies of cytotoxic lymphocytes have involved the characterisation of recognition structures on cytotoxic lymphocytes and the definition of two mechanisms of cytotoxicity. In contrast to normal cell death, which occurs during embryonic development and the formation and death of hematopoietic cell lineages, lymphocyte-mediated cytotoxicity occurs in the context of an inflammatory response and the dying cells are lysed into the surroundings rather than phagocytosed. The roles of the two different forms of lymphocyte-mediated cytotoxicity are quite distinct; however they both involve induction of an endogenous pathway of apoptosis in the targeted cell, and they do share features with all other forms of physiological cell death. PMID- 7487971 TI - Cytomegalovirus and atherosclerosis. AB - Finding that an avian herpesvirus can cause atherosclerosis in chickens prompted studies of human herpesviruses in human atherosclerosis. Antigens and nucleic acid sequences of cytomegalovirus (CMV), a widespread member of the herpesvirus family, were found in arterial lesions in human atherosclerosis, but infectious virus has not been observed. In atherosclerosis patients, high levels of CMV antibodies are present, suggesting the presence of virus that had been activated from a latent state. Atherosclerosis also develops in immune-suppressed heart transplant patients infected with CMV. The properties of CMV are consistent with its involvement at several levels of the atherogenic process. If this concept is correct, immunization with a CMV vaccine should prevent CMV infection and atherosclerosis. PMID- 7487972 TI - Variation in the human genome. 12-15 June, 1995. CIBA Foundation symposium 197, London, UK. PMID- 7487973 TI - Induction and the developmental zeitgeist. 14-17 JUne, 1995; Aland, Finland. PMID- 7487974 TI - Ribozymes cleave the AML1/MTG8 fusion transcript and inhibit proliferation of leukemic cells with t(8;21). AB - The AML1/MTG8 fusion gene is thought to have a critical role in the leukemogenesis of AML with t(8;21)(q22;q22). To specifically inhibit the proliferation of leukemic cells having the AML1/MTG8 fusion gene, we constructed two hammerhead ribozymes against AML1/MTG8. Two cleavage sites were targeted as follows: site 1 for ribozyme 1(Rz1), a CUC located 3 bases upstream from the fusion site; site 2 for ribozyme 2(Rz2), an AUC located 3 bases downstream from the fusion site. In a cell-free system, Rz1 and Rz2 specifically cleaved AML1/MTG8 substrate, dependent on the concentration of ribozymes. When these ribozymes were transfected to Kasumi-1 cells, an AML cell line with AML1/MTG8, they were able to inhibit the cell growth. These data suggest that Rz1 and Rz2 may be applied as a new therapeutic agent in the treatment of AML with t(8;21). PMID- 7487975 TI - Integrin stimulation decreases tyrosine phosphorylation and activity of focal adhesion kinase in thymocytes. AB - FAK, focal adhesion kinase, is expressed in a variety of cell types and has been suggested to transduce signals brought about by integrin-extracellular matrix (ECM) interactions. Integrin stimulation increases tyrosine phosphorylation and activity of FAK in all the cells examined to date. In contrast, in thymocytes stimulation of VLA-4 (alpha 4 beta 1) and LFA-1 (alpha L beta 2) resulted in a marked decrease in tyrosine phosphorylation and activity of FAK. PMID- 7487976 TI - Consequences of p53 gene expression by adenovirus vector on cell cycle arrest and apoptosis in human aortic vascular smooth muscle cells. AB - p53 shows its tumor suppresser activity by inducing cell cycle arrest and/or apoptosis of tumor cells and these activities are in part mediated by p21 cyclin dependent kinase inhibitor (also called as WAF1, Cip1 and SDI1). Using human aortic vascular smooth muscle cells, here we demonstrate that adenovirus vector expressing p53-induced p21, cell cycle arrest at G1 and G2/M boundary, and accumulation of cells in G1 subgroup. However, adenovirus vector expressing p21 induced only G1 cell cycle arrest. The adenovirus vector expressing p53 was 200 times more cytotoxic to human aortic vascular smooth muscle cells than adenovirus vector expressing p21. These results suggest that adenovirus expressing p53 induces cytotoxicity in human vascular smooth muscle cells by apoptosis and this cytotoxicity can not be fully accounted by p21 induction. PMID- 7487977 TI - Intracellular stability of alpha fragments of beta-galactosidase: effects of amino-terminally fused polypeptides. AB - Intracellular stability of alpha fragments of beta-galactosidase in Escherichia coli has been studied by pulse-chase/immunoprecipitation experiments. An alpha fragment encoded by the pUC118 vector was relatively stable with an estimated half-life of about 12 min at 37 degrees C, whereas another vector, pSTV28, encoded a less stable alpha fragment that had a different carboxy-terminal sequence. Stability of the fragment was found to be affected markedly by amino terminal attachment of other sequences. An amino-terminal fusion of a sequence derived from cytoplasmic domain 4 of the SecY protein shortened the half-life of the alpha fragment to less than 1 min. In contrast, an amino-terminal sequence from the NusG protein had no apparent effect on the stability of the fragment. In a fusion protein in which the intact SecY protein was fused to the alpha fragment, stabilization of the SecY part by overproduction of the partner SecE protein resulted in an increased alpha complementation activity of beta galactosidase. These results indicate that stability of alpha fragment can be dictated by the stability of the fused protein. The alpha fragment of beta galactosidase, which is unique in that it is largely unstructured but can be "active" in alpha complementation, may be used as an in vivo indicator of stability of proteins attached to it. PMID- 7487978 TI - Spin-lattice relaxation times of water in polarized and depolarized rabbit vagus nerves. AB - Spin-lattice relaxation times (T1) of intracellular water of the nonmyelinated fibers of rabbit cervical vagus nerve were measured using a paramagnetic shift reagent, s-FDF. Spin-lattice relaxation decay curves were composed of the fast (T1,F) and slow (T1,S) relaxation components. The mean values of T1,F and T1,S in the polarized nerve fibers at 25 degrees C were 0.12 +/- 0.04 and 0.61 +/- 0.13 sec., respectively, and their fractions were 0.71 +/- 0.07 and 0.29 +/- 0.07, respectively (n = 21). On the other hand, those values in the depolarized nerve fibers were 0.16 +/- 0.01 sec., 0.77 +/- 0.17 sec., 0.77 +/- 0.10 and 0.23 +/- 0.10, respectively (n = 15). T1,F and T1,S for the depolarized nerve fibers were significantly elongated (P < 0.01). PMID- 7487979 TI - Nuclear location of the regulatory protein of glucokinase in rat liver and translocation of the regulator to the cytoplasm in response to high glucose. AB - Rat liver is known to contain a regulatory protein (RP) that inhibits glucokinase competitively with respect to glucose. By an immunohistochemical technique and by cell fractionation in glycerol, we investigated the distribution of RP in rat liver and its changes in response to high glucose. RP was detected almost exclusively in the nuclei of the parenchymal cells of both fed and fasted rats. In situ perfusion of the liver with 20 mM glucose for 10 min caused a marked decrease in nuclear RP and an increase in cytoplasmic RP. These results indicate that RP is present predominantly in the nuclei of hepatocytes and is translocated from the nucleus to the cytoplasm in response to high glucose. PMID- 7487980 TI - Interaction of peptide substrates of fibroblast collagenase with divalent cations: Ca2+ binding by substrate as a suggested recognition signal for collagenase action. AB - To correlate structural data on substrates of human fibroblast collagenase with their interaction with the enzyme, we have studied: Ac-PLG-s-LLG-O-ethyl ester (I), Dnp-PLGLWA(d-Arg)-NH2 (II), AcGPEGLRVG-O-ethyl ester (III) and Succ-GPLGP-O amidomethylcoumaryl ester (IV). Peptides I and II represent collagenase cleavage sequences in collagen, peptide III is a mimic for the cleavage site in alpha 2 macroglobulin and peptide IV represents a non-substrate model. Kinetic data showed that peptides I, II and III were substrates of the enzyme. In contrast, peptide IV was not acted upon by the enzyme. Circular dichroism data on the peptides showed that the peptides assume ordered structures in water and trifluoroethanol. In the latter solvent, peptides I and III bound Ca2+ and Zn2+ while peptide II bound Ca2+ but not Zn2+. Peptide IV did not bind either cation in this solvent. Together with the kinetic data, the results suggest that the collagenase cleavage segments in collagen and non-collagen substrates of collagenase could interact with Ca2+ and the enzyme to form a ternary complex. This, in turn, would imply a cofactor role for Ca2+ in collagenase action in addition to the solely structural role ascribed so far to this cation. PMID- 7487981 TI - Expression of local hepatocyte growth factor system in vascular tissues. AB - Since endothelial cells (EC) are known to secrete various anti-proliferative and vasodilating factors, an agent that promotes seeding or regeneration of EC may have potential therapeutic value against vascular smooth muscle cell (VSMC) proliferation. For the above purpose, we have found that hepatocyte growth factor (HGF) fulfills such conditions. However, the local HGF system has not yet clarified. Therefore, we investigated endogenous HGF production and its specific receptor (c-met) in vascular tissues. Our results revealed the expressions of HGF and its receptor (c-met) mRNA in endothelial cells, VSMC and neointimal VSMC of rat assessed by RT-PCR. HGF and c-met mRNA were also detected in human aortic endothelial cells and VSMC. Endothelial cells and VSMC of both rat and human produce and secrete immunoreactive HGF as assessed by ELISA. Of importance, the existence of local HGF system (HGF and c-met) was also observed in vivo in intact abdominal aorta of SD and Wistar rats. Overall, this study demonstrated that HGF is expressed and secreted from endothelial cells and VSMC of both rat and human in vitro as well as in vivo. Since HGF has characteristics of an endothelium specific growth factor, locally synthesized HGF in endothelial cells and VSMC may have a role in vascular functions in autocrine-paracrine manner. PMID- 7487983 TI - Characterization of the murine PIG-A promoter region high constitutive PIG-A gene expression in brain. AB - PIG-A is an X-linked gene that encodes a synthetic element required to initiate glycoinositol phospholipid (GPI) anchor assembly. In this study we characterized genomic sequence flanking the previously identified 5' end of the murine PIG-A gene and analyzed constitutive levels of PIG-A gene expression in vivo in different tissues. We found that the 5'-flanking sequence contains 1) alternative transcriptional start sites at -154 and -34/-33 relative to previously known exon 1 sequence, 2) an MRE and multiple GRE, Ap-2 and Sp-1 consensuses, and 3) a 100 bp-long GC-rich segment bounded by NF-IL6 sites. Our survey of PIG-A mRNA levels in vivo unexpectedly revealed that PIG-A expression levels are 2-4 fold higher in the brain than in other tissues. PMID- 7487982 TI - Tumor necrosis factor alpha increases tyrosine phosphorylation of a 23-kDa nuclear protein in U937 cells through ceramide signaling pathway. AB - In many tumor cell lines, tumor necrosis factor alpha (TNF alpha) causes profound alterations in nucleus including chromatin condensation and DNA fragmentation. However the mechanism is largely unknown. Here we demonstrate that TNF alpha treatment of U937 cells increased tyrosine phosphorylation of a 23-kDa nuclear protein (P23) maximally by 13-fold, which occurred after 2.5 h treatment concomitantly with occurrence of DNA fragmentation. Tyrosine kinase inhibitor, herbimycin A, inhibited either tyrosine phosphorylation of P23 or DNA fragmentation, suggesting that the P23 phosphoprotein may be involved in the TNF alpha-induced changes in nucleus. Increase of intracellular Cer level by hydrolysis of cell surface SM with bacterial sphingomyelinase or addition of either C2-Cer or natural Cer also resulted in increase of P23 tyrosine phosphorylation, which was detectable at 1.5 h treatment and reached maximum after 2 h treatment, indicating that P23 phosphoprotein may be a downstream effector of Cer signaling pathway. Our data provide further evidence that Cer signaling pathway may mediate signal transduction of TNF alpha. PMID- 7487984 TI - Evidence for a transient interresidue hydrogen bond in sucrose in aqueous solution obtained by rotating-frame exchange NMR spectroscopy under supercooled conditions. AB - We have used the hydroxyl protons of sucrose dissolved in supercooled water as NMR probes for the detection of intramolecular hydrogen bonds. Neither the measured OH temperature shift coefficients, 3JHCOH scalar couplings, isotope induced 13C chemical shifts (delta delta COH/COD), nor OH exchange rates allowed us to single out any hydroxyl group in sucrose with characteristics indicative of involvement in strong hydrogen bonding. However, two-dimensional rotating-frame exchange (ROESY) spectroscopy revealed a direct exchange between the glucosyl OH2 and fructosyl OH1 protons. We conclude that an O2g::H::O1f interresidue hydrogen bond transiently exists in sucrose in aqueous solution. ROESY of sugars in supercooled water is proposed as a novel method to detect this type of weak hydrogen bonding. PMID- 7487985 TI - Down-regulation of bcl-xs gene expression in rat thymocytes by dexamethasone. AB - In order to test a hypothesis that the expression of bcl-x gene promotes apoptosis by antagonizing the function of bcl-2 gene product, time course of the expression of bcl-x was investigated using the dexamethasone-treated rat thymocytes which were undergoing apoptosis. Unexpectedly, dexamethasone suppressed the expression of bcl-x in a dose- and time-dependent manner. Such decrease was detected even in the presence of cycloheximide, suggesting that the suppression is due to the genomic (primary) effect of dexamethasone. Since the expression of bcl-xL, as measured by a specific nucleotide probe, was not detected at a quantifiable level, the decrease was apparently attributed to that in bcl-xS. Our present observations propose a new role of the bcl-xS gene product, in which the bcl-x gene product may be necessary for survival of immature thymocytes rather than for their apoptosis. PMID- 7487986 TI - cDNA cloning and bacterial expression of the human type I keratin 16. AB - The human type I keratin 16 is constitutively expressed in a number of complex epithelial tissues, including skin, but is better known for its induction under conditions favoring enhanced proliferation or abnormal differentiation, including wound healing, psoriasis, and cancer. We cloned the coding sequence of human K16 by applying a coupled reverse transcription-polymerase chain reaction procedure to mRNAs prepared from cultured human skin keratinocytes. We then expressed the human K16 coding sequence in E. coli and purified the solubilized protein by anion-exchange chromatography. The recombinant protein recovered behaves similarly to human K14 (a related acidic keratin) on the anion-exchanger, co migrates with native human K16 on SDS-PAGE (M(r) 48 kD), and reacts with antisera directed against human K16. Based on the nucleotide sequence obtained and the properties of the corresponding recombinant protein, we conclude that we have cloned the coding portion of the human K16 cDNA. The sequence data obtained in this study is compared to earlier reports of the human K16 sequence, which are conflicting in many respects. The availability of K16 in a purified recombinant form will allow us to study how its properties may relate to its function during wound healing and in skin diseases. PMID- 7487987 TI - Nicotinamide inhibits IRF-1 mRNA induction and prevents IL-1 beta-induced nitric oxide synthase expression in pancreatic beta cells. AB - Nitric oxide produced by inducible nitric oxide synthase in islets exerts inhibitory and cytotoxic effects on pancreatic beta cells and is therefore thought to be a potent mediator in the pathogenesis of Type I diabetes mellitus. Here, using isolated rat pancreatic islets, we show that high-concentration nicotinamide (20 mM), but not low-concentration nicotinamide (5 mM), attenuates the interleukin-1 beta-evoked inhibition of glucose-induced insulin secretion by preventing the induction of interferon regulatory factor-1, a transcriptional factor which plays an essential role in inducible nitric oxide synthase gene expression, and the interleukin-1 beta-induced nitric oxide formation. High concentration nicotinamide also restored an interleukin-1 beta-induced decrease in ATP content in pancreatic beta cells, suggesting that interleukin-1 beta induced nitric oxide inhibits the mitochondrial function. The present results show the molecular basis of the preventive effect of high-dose nicotinamide on Type I diabetes mellitus. PMID- 7487989 TI - Involvement of PIM-1 in DNA fragmentation in mouse NS-1-derived cells. AB - In several cell lines derived from mouse NS-1 myeloma cells, internucleosomal fragmentation of chromosomal DNA, a hallmark of apoptosis, was continuously observed. Approximately 15-20% of the cells died when isolated in a 96-well plate, and the surviving cells contained fragmented DNA ('ladder'). Among a variety of genes so far reported to be related to apoptosis, only Pim-1 was expressed at an elevated level in the NS-1 hybridomas as compared in a control myeloma cell line without 'ladder'. Transfection of a Pim-1 expression vector to a 'ladder'-non-producing myeloma line yielded similar internucleosomal DNA fragmentation. The results hence suggested that Pim-1 activates endonucleases responsible for DNA fragmentation during apoptosis and/or repress DNA repair systems to restore fragmented DNA. PMID- 7487988 TI - Endotoxin markedly elevates plasma concentration and gene transcription of adrenomedullin in rat. AB - A marked increase in adrenomedullin (AM) concentration was observed in rat plasma collected after intravenous infusion of lipopolysaccharide (LPS). Plasma immunoreactive (ir-) AM concentration increased already after 1 h exposure to 5 mg/kg of LPS and was elevated to 49.7 +/- 2.3 fmol/ml (mean +/- SEM) at 3 h after injection in contrast to 2.7 +/- 0.3 fmol/ml in saline-injected control rats. Plasma ir-AM level increased in a dose-dependent manner in a range of 0.008 to 5.0 mg/kg of LPS. AM gene transcription in LPS-injected rats was augmented more than 3-fold in ileum, liver, lung, aorta, and 2.4-fold even in skeletal muscle, in which AM was not thought to be produced in myocytes. These results, along with our recent data that AM production in cultured vascular smooth muscle cells (VSMCs) is stimulated with LPS, indicate that AM production is highly augmented in blood vessel, lung and intestine by administration of LPS. Since VSMCs express AM-specific receptors, secreted AM is deduced to actually exert a vasorelaxant effect especially in the endotoxin shock. PMID- 7487990 TI - Sialyl Lewis X moiety on rat polymorphonuclear leukocytes responsible for binding to rat E-selectin. AB - We investigated the presence of the carbohydrate ligand for E-selectin on the cell surface of rat polymorphonuclear leukocytes (PMN). Rat PMN, isolated from peripheral blood, adhered to recombinant rat E-selectin-coated microplates. The adhesion was inhibited either by an anti-rat E-selectin monoclonal antibody (MAb), a sialyl Lewis X (SLex) oligosaccharide or neuraminidase digestion. FACS analysis revealed the expression of SLex as well as other adhesion molecules such as L-selectin, CD11a, CD11b and CD18 on the cell surface of rat PMN. The binding of an anti-SLex MAb KM93 to rat PMN was inhibited competitively by a SLex but not by a Lewis X (Lex). The reactivities of two anti-SLex MAbs, KM93 and CSLEX-1, or an anti-Lex MAb BC90/45 to rat PMN differed from those to human PMN. These results suggest that rat PMN contain the SLex-moiety, which binds to rat E selectin and is different from that of human PMN. PMID- 7487991 TI - A mutation in the beta 3-adrenergic receptor gene is associated with obesity and hyperinsulinemia in Japanese subjects. AB - The Trp 64 Arg mutation in the beta 3-adrenergic receptor (beta 3AR) gene was investigated in 350 Japanese subjects. This mutation was not associated with non insulin-dependent diabetes mellitus (NIDDM). In 191 subjects without NIDDM, body mass index (BMI) was significantly higher in subjects homozygous for this mutation than in those homozygous for the normal allele (24.7 +/- 1.4 vs 22.1 +/- 0.2 kg/m2, p = 0.009). Moreover, the frequency of the mutant allele in obese subjects (BMI > 26.4) was significantly higher than that in non-obese subjects (BMI < 22) (0.37 vs 0.15, p = 0.009). The presence of this mutation was also accompanied by significantly higher fasting (p = 0.000) and 2 hrs (p = 0.018) serum insulin levels during an oral glucose tolerance test. The beta 3AR may be one of the loci contributing to obesity and hyperinsulinemia/insulin resistance in Japanese subjects. PMID- 7487993 TI - Effect of KN-62, Ca2+/calmodulin-dependent protein kinase II inhibitor, on adriamycin resistance of human ovarian cancer cells. AB - We examined effects of an isoquinolinesulfonamide derivative, KN-62, on human ovarian cancer cells, NOS3AR, that are resistant to Adriamycin (ADR). MTT assay revealed that 10 microM KN-62 overcame the resistance. KN-62 had little effect on GST activity. In studies on the intracellular accumulation of ADR, KN-62 increased the ADR contents in the resistant cells close to the level seen in the sensitive cells. These results suggest that the reversal of the resistance against ADR in ovarian cancer cells by KN-62 is mainly due to higher accumulation of ADR in NOS3AR cells. Furthermore, we detected Ca2+/calmodulin-dependent protein kinase II (CaM kinase II) in NOS3AR cells since KN-62 is a specific inhibitor of the kinase. In this paper, we discussed on modulation of ADR resistance by KN-62. PMID- 7487992 TI - Evidence for a glycyl-proline transport system in ovine enterocyte brush-border membrane vesicles. AB - A peptide transport system in ovine enterocyte BBMV has been examined with glycyl L-proline (Gly-Pro) as substrate. Transport is stimulated by an inwardly directed proton gradient (pH1 = 8.4; pH0 = 6.0). Gly-Pro uptake was saturable and conformed to Michaelis-Menten kinetics. Uptake was not affected by the presence of glycine but was inhibited by glycyl-L-phenylalanine (Gly-Phe). This is thought to be the first positive demonstration of the existence of a peptide transporter in the ruminant small intestine. PMID- 7487995 TI - A single base-pair change (ATG-->ATC) nullifies the activity of cytosolic fumarase in Saccharomyces cerevisiae. AB - A respiratory defective pet mutant (W303 delta FUM1) of Saccharomyces cerevisiae deficient in fumarase was transformed with the plasmid construct pG5/ST7. This plasmid contains the entire FUM1 gene with only a single base pair change (ATG- >ATC) confined to the putative second inframe translation initiation codon. While transformation of the fumarase deficient mutant with pG5/ST7 resulted in an elevation of fumarase activity in the mitochondria of the transformed strain, fumarase activity in the cytosol remained negligible his result indicated that the cytosolic fumarase isoenzyme is synthesized exclusively from the second translation initiation codon of FUM1. Results of RACE-PCR of the 5' ends of FUM1 transcripts confirmed that the FUM1 gene synthesizes two distinct species of transcripts. These data provide strong evidence for the synthesis and targeting of two fumarase isomers. PMID- 7487994 TI - Selective killing of cholinergic neurons by microglial activation in basal forebrain mixed neuronal/glial cultures. AB - Microglia activation by lipopolysaccharides (LPS) significantly decreased choline acetyltransferase-immunopositive (ChAT+) neuron number and ChAT activity in rat primary basal forebrain mixed neuronal/glial cultures. The number of non cholinergic (ChAT(-)) neurons was unaffected. LPS induced nitric oxide synthase (NOS) in microglia, increased the media level of the NO metabolite nitrite, and the NOS inhibitor Ng-nitro-L-arginine methylester (NAME) protected the ChAT+ neurons from LPS. The combination of beta-amyloid peptide (1-42) and interferon gamma (INF-gamma) also increased the media nitrite level and decreased ChAT+ neuron number. Cholinergic neurons are lost early in the course of Alzheimer's disease, and the enhanced sensitivity of these neurons to microglial activation in mixed neuronal/glial culture may be useful for modeling Alzheimer's disease and developing therapeutic strategies to combat this disease. PMID- 7487996 TI - Molecular cloning and functional analysis of the rat mu opioid receptor gene promoter. AB - The rat mu opioid receptor gene promoter was cloned and characterized. It has a few features in common with the mouse gene, e.g. the lack of a classical TATA box and the fact that several transcriptional start sites are used. The overall homology between the two species is greater than 85%. Functional analysis of the promoter was performed using transient expression of rat mu opioid receptor reporter gene constructs in the mu opioid receptor expressing cell line SH SY5Y and in non mu opioid receptor expressing cell lines. A promoter region was defined which confers both high basal and TPA and forskolin stimulated reporter gene expression in SH SY5Y cells. PMID- 7487997 TI - Nuclear matrix condensation and c-myc and c-fos expression are specifically altered in culture rat hepatocytes after exposure to cyproterone acetate and phenobarbital. AB - Regenerative or hyperplastic growth promotes carcinogenesis and can be induced by many nongenotoxic carcinogens. The mitogenic potential of the rodent liver tumor promoters, cyproterone acetate and phenobarbital was investigated in primary rat hepatocyte cultures. Two premitotic markers were analyzed, the expression of two immediate-early genes (c-fos and c-myc) and the decrease in the nuclear quinacrine dihydrochloride fluorescence indicative for a G0-G1 cell cycle shift. C-fos expression and decrease in nuclear fluorescence could be induced by both chemicals, phenobarbital being the lesser potent, whereas c-myc expression was only inducible by cyproterone acetate. In situ hybridization with c-myc revealed that both chemicals enhanced c-myc mRNA levels in individual cells, however the number of responding hepatocytes was increased by cyproterone acetate only. The chemical-induced premitotic changes in hepatocytes were highly specific in terms of affected genes and ploidy levels of responding hepatocytes. PMID- 7487998 TI - Functional analysis of the 5' flanking sequence in the ovine beta 1-adrenergic receptor gene. AB - Functional data for the promoter of the beta 1-adrenergic receptor (beta 1AR) gene are lacking. We previously cloned the ovine beta 1AR gene and mapped the transcription start sites. We now report data on ovine beta 1AR gene expression obtained by transient transfection. Progressive deletion of upstream 5' flanking region moderately increased transcription activity in three cell lines compared to the full-length promoter. Deletion of sequences between -1530 and -953 produced the greatest increase in transcriptional activity. This region encompassed a putative GRE and an AP1 site. Deletion of the transcription start sites eliminated nearly all of the activity. Dexamethasone significantly increased activity of each of the promoter constructs tested in C6 glioma cells and an embryonic myocardial cell line, W1 cells. T3 alone had no effect and cotreatment with T3 did not augment the effects of dexamethasone. We conclude, basal transcription activity is repressed by a mechanism which operates through element(s) in the proximal promoter. Glucocorticoids increase transcription through mechanism(s) within the same region. We speculate that this region in the ovine beta 1AR promoter may be responsible for its unique transcription regulation. PMID- 7487999 TI - N-acetylcysteine diminishes injury induced by balloon angioplasty of the carotid artery in rabbits. AB - The effect of N-acetylcysteine (NAC) on preventing or ameliorating the injury associated with percutaneous transluminal angioplasty was investigated in rabbits. Carotid artery angioplasty (CA) was performed on 8 control (vehicle treated) rabbits and 7 rabbits treated with NAC, 250 mg/kg, administered orally in Nutrical paste for 10 days prior to and 10 days following CA. Single blind histologic evaluation of the angioplasty sites demonstrated a significant reduction in the incidence in vessel: 1) inflammation; 2) endothelial damage; 3) thrombus formation; 4) elastic lamina damage in the NAC rabbits. The percentage of control versus NAC treated animals exhibiting damage in each category were: inflammation: 75% vs 14%; endothelial damage: 88% vs 57%; thrombus formation: 88% vs 43%; laminal damage: 63% vs 14%. The results suggest that NAC treatment may be a valuable therapeutic agent in effectively preventing or reducing angioplasty induced vessel damage. PMID- 7488000 TI - Autocrine regulation of the renal arterial tone by adrenomedullin. AB - We tested the hypothesis that adrenomedullin (ADM), a recently isolated hypotensive peptide, is an autocrine transmitter in renal arterial smooth muscle cells (RASMCs). Fura-2 fluorometry revealed that ADM decreases both the cytosolic Ca2+ concentration ([Ca2+]i) and tension induced by 0.3 microM phenylephrine (PE) in porcine RASMCs. The expression of ADM mRNA in RASMCs was assessed by the reverse transcription polymerase chain reaction (RT-PCR), using the total RNA from the RASMCs and the specific primers for the porcine ADM mRNA. A band with an expected size could be detected only when the reverse transcriptase was added, thus indicating that the porcine RASMCs express ADM mRNA. These results therefore suggested that ADM plays an important role in the regulation of the renal vascular tone, not only as a circulating hormone, but also as an autocrine transmitter. PMID- 7488001 TI - The synthetic [Tyr5,12,Lys7]-polyphemusin II peptide (T22) binds to the CD4 cell surface molecule. AB - The [Tyr5,12,Lys7]-polyphemusin II peptide (T22) inhibits HIV-1 replication in lymphocytes. The mechanism of T22 inhibition of HIV-1 replication may involve T22 competition with HIV-1 for attachment sites on the plasma membrane of targeted cells. Here we find that the T22 peptide binds to the CD4 molecule in affinity columns. We also find that antiserum to CD4 inhibits cell attachment to T22. Further CD4+ transfected cells attach to T22 while their parental cells which do not express CD4 do not attach to T22. These data demonstrate that T22 binds to the CD4 molecule and supports the hypothesis that T22 inhibits HIV-1 replication by binding to the cell surface CD4 molecule and inhibiting uptake of the virus. PMID- 7488002 TI - Purification and characterization of zeta-crystallin from the camel lens. AB - zeta-crystallin a novel NADPH: quinone oxidoreductase was purified from the cortex of the camel (Camelus dromedarius) lens to homogeneity by Sepharose CL-6B gel filtration column and 2', 5' ADP-Sepharose 4B affinity column chromatography in the presence of dithiothreitol. The purified zeta-crystallin has a molecular weight of 140 kDa, as determined by Superose 12 gel filtration column. SDS-PAGE showed a single polypeptide band of molecular weight 35 kDa, suggesting that the native enzyme is composed of four identical subunits. The isoelectric point of the enzyme was 7.6 on native polyacrylamide gel. The enzyme was purified 20-fold over homogenate with a specific activity of 26.0 Units/mg protein, and an overall recovery of 53%. This enzyme was NADPH specific and followed Michaelis-Menten kinetics. Km values for the reduction of 9,10-phenanthroquinone and oxidation of NADPH were 17 microM and 6.9 microM, respectively, at pH 7.8. The Vmax values of the enzyme for 9,10 phenanthroquinone and NADPH were 32 mumole min-1, mg-1 protein and 22.7 mumole min-1 mg-1 protein, respectively. PMID- 7488003 TI - Phorbol esters attenuate the expression of p53 in cells treated with doxorubicin and protect TS-P53/K562 from apoptosis. AB - The induction of p53 by doxorubicin in normal human fibroblasts was completely reverted by TPA, a phorbol ester. A ts-p53 mutant protein which is ineffective at 37 degrees C, but behaves in a wild-type fashion at 32 degrees C, was overexpressed in the p53-null human leukemia cell line K562. Wild-type-p53 overexpression induced apoptosis, whereas TPA protected K562 cells from this phenomena. By analogy with the observed human fibroblasts, TPA was found to decrease p53 amount. The TPA-dependent down-regulation of p53 could explain the chromosomal gross alterations typical of cells subjected to a chronic TPA treatment, alterations also found in cells defective for p53 function. PMID- 7488004 TI - Molecular cloning and in vivo expression of the bovine steroidogenic acute regulatory protein. AB - Differential screening was used to select clones from a bovine luteal cDNA library, which were specifically expressed in late luteal stages. One of these clones encoded the bovine homologue of the Steroidogenic Acute Regulatory protein (StAR). The bovine StAR gene is transcribed as 3kb and 1.8kb transcripts, which differ in their sites of 3' polyadenylation. Both transcripts are highly expressed in corpora lutea of mid to late cycle and through pregnancy, but only at low levels in the early cycle. Northern analysis showed expression only in the corpus luteum and in the adrenal gland, but not in any other tissue examined. Within the protein coding region of the bovine StAR gene, there is a marked 124 base homology to the 5' non-coding region of another luteal transcript, TIMP-1, suggesting a possible common regulatory function for this sequence. PMID- 7488005 TI - Mechanical and thermal changes in the Torpedo electric organ associated with its postsynaptic potentials. AB - Rapid mechanical and thermal changes in slices of the Torpedo electric organ evoked by electric stimulation were investigated. The organ was found to swell simultaneously with the postsynaptic potential. This swelling was followed by prolonged shrinkage of the organ. These findings may be explained from the proven facts that the acetylcholine(ACh)-receptor proteins have a large binding capacity for Ca-ions and that addition of ACh to the medium can cause release of Ca-ions from the receptor proteins. PMID- 7488006 TI - Expression of ras gene in flounder (Platichthys flesus) andred mullet (Mullus barbatus). AB - A member of the ras gene family was identified here for the first time in two marine fish, the flounder (Platichthys flesus) and the red mullet (Mullus barbatus). After RT-PCR the partial sequence of ras gene (exons 1 and 2) has been cloned and sequenced from normal liver. At the nucleic acid level, a very high extent of homology with the human ras genes is observed (80% to 86%) and a perfect homology is observed at the amino acid level. The high conservation of the 5' region of the ras gene suggest that the p21 protein has identical function in higher and lower vertebrates. The identification of ras gene should be a first step in understanding the molecular basis of carcinogenesis in these species. PMID- 7488007 TI - Molecular cloning and developmental regulation of expression of two isoforms of the catalytic subunit of protein phosphatase 2A from Xenopus laevis. AB - Two types of PP2A catalytic subunit cDNA clones were isolated from a Xenopus oocyte library. One type corresponds to the Xenopus C36 alpha (XC36 alpha) isoform as published by Cormier et al. (1991) and another type encodes for a novel isoform of PP2AC36 (XC36 beta), more homologous to the C36 beta isoform cloned from Mammalia. Northern blot analysis with isoform specific probes showed that the XC36 alpha mRNA transcript of 2 kb is more abundant than the XC36 beta mRNA of 1.9 kb during embryogenesis and in adult tissues. Both transcripts are highly expressed in ovary and heart relative to the other adult tissues examined. The high expression levels of both XC36 alpha and XC36 beta transcripts in the ovary decrease during the first mitotic embryonic divisions. Resumption of zygotic expression starts at about the same time for both mRNAs, during the tailbud stage, but the XC36 beta transcript shows only a moderate and transient increase and declines again during the tadpole stage whereas expression of XC36 alpha increases further during the same period until the onset of metamorphosis. PMID- 7488008 TI - Decreased beta-adrenergic stimulation of glycoprotein secretion in CF mice submandibular glands: reversal by the methylxanthine, IBMX. AB - beta-adrenergic stimulation of glycoprotein secretion was shown to be decreased in submandibular glands of Cystic Fibrosis (CF) mice. The defective response was partially restored by the methylxanthine, IBMX or cpt-cyclic AMP. Cholinergic stimulation of pancreatic amylase secretion was not affected in CF mice, demonstrating that this is not a generalised depression of protein secretion. The data are the first to show that the CF mouse mimics the protein secretion defect in CF human submandibular cells and that the mechanism of correction of the CF defect is via elevation of cyclic AMP. The results are therefore invaluable towards devising a rational pharmaceutical therapy for CF patients. PMID- 7488010 TI - Secretion of prostaglandins elicited by lipopolysaccharide and ethanol in cultured rat Kupffer cells. AB - Prostaglandins (PGs) released by cultured rat Kupffer cells in response to stimulation with lipopolysaccharide (LPS) or ethanol were extracted from culture media, separated by HPLC and measured by radioimmunoassay. LPS (0.5-5 micrograms/ml) enhanced, after a 3-4 hrs lag period, the production of PGE2 (7-10 fold by 24 hrs), thromboxane B2 (2-3 fold) and PGD2. PG 6-keto-F1 alpha, PGF2 alpha (20-50% each). This effect was not inhibited by 30 microM aspirin but was reduced by dexamethasone. Ethanol (25-85 mM) gradually increased the release of PGE2 (40-90% by 24 hrs) and other PGs (10-30%), with 30 microM aspirin eliminating this effect. When added together with LPS, ethanol potentiated the endotoxin action. We suggest that LPS causes synthesis of the inducible cyclooxygenase-2 form in Kupffer cells, whereas ethanol exerts its effect via the pre-existing cyclooxygenase-1 mainly by increasing the free arachidonic acid content. PMID- 7488009 TI - The carboxyl extensions of two rat ubiquitin fusion proteins are ribosomal proteins S27a and L40. AB - Two rat recombinant cDNAs were characterized; they encode fusion proteins that have at their NH2 terminus the conserved 76 amino acid ubiquitin and at their carboxyl terminus the extension ribosomal proteins S27a (80 amino acids and a molecular weight of 9,397) or L40 (52 amino acids and a molecular weight of 6,177). The fusion proteins are processed in a reticulocyte lysate to ubiquitin and either S27a or L40. Hybridization of cDNAs to digests of nuclear DNA suggests that there are 14 to 19 copies of the S27a, and 6 to 10 of the L40, genes. The mRNA for ubiquitin-S27a has about 700 nucleotides and ubiquitin-L40 about 650. Ribosomal proteins S27a and L40 contain zinc finger motifs of the C2-C2 variety. S27a and L40 are related to ribosomal proteins from other species. PMID- 7488011 TI - Increased production of hydroxyl radical by pericentral microsomes compared to periportal microsomes after pyrazole induction of cytochrome P4502E1. AB - Cytochrome P4502E1 is localized in the pericentral (PC) zone of the liver acinus to a greater extent than in the periportal (PP) zone. After pyrazole treatment, PC microsomes were more active in oxidizing typical substrates of CYP2E1 than PP microsomes and had an increased content of CYP2E1. The ability of PC and PP microsomes from pyrazole-treated rats to interact with iron and generate reactive oxygen species such as the hydroxyl radical (.OH) was evaluated. A sensitive DNA strand cleavage assay was used to detect .OH; supercoiled plasmid DNA is compact but is converted by .OH-induced single strand breaks to the relaxed open circular state. Microsomes from PC hepatocytes of pyrazole-treated rats were several fold more reactive than PP microsomes in promoting NADPH-dependent DNA strand cleavage with a variety of iron catalysts, including ferric-ATP, ferric-histidine, ferric citrate, ferric ammonium sulfate, and ferric-EDTA. DNA strand cleavage was inhibited by superoxide dismutase, catalase, and .OH scavengers such as DMSO and ethanol. Rates of H2O2 production were higher with the PC microsomes. These results indicate that rates of .OH production are higher with PC microsomes than PP microsomes after pyrazole treatment to induce cytochrome P4502E1 and suggest the possibility that elevated production of reactive oxygen species may play a role in ethanol toxicity to the PC zone of the liver acinus. PMID- 7488013 TI - Retinoic acid induces three newly cloned HOXA1 transcripts in MCF7 breast cancer cells. AB - Coordinated expression of genes involved in development, differentiation and malignant transformation is regulated by transcription factors including homeodomain-containing proteins. However, most of their cDNA sequences are still unknown. We report here the molecular characterization of three newly cloned HOXA1 transcripts from human breast cancer cells. In addition, we provide evidence that these alternatively spliced transcripts encode one homeodomain containing protein and two products lacking the conserved DNA-binding domain. Moreover, we demonstrate that all three HOXA1 transcripts are induced by retinoic acid in MCF7 cells. Taken together, our results suggest that HOXA1 gene may be a key element in the establishment of the breast cancer cell phenotype. PMID- 7488014 TI - Serine and cysteine proteinase inhibitors prevent nitric oxide production by activated macrophages by interfering with transcription of the inducible NO synthase gene. AB - The objective of this study was to ascertain the mechanism by which serine and cysteine proteinase inhibitors interfere with production of NO by LPS-activated rat alveolar macrophages. Macrophages were incubated in the presence of LPS+ test agent for 24 hr. Culture media were analyzed for NOX- accumulation, harvested cells were assayed for iNOS activity, and cellular RNA was extracted for determination of iNOS mRNA by Northern blot analysis. TPCK, TLCK, calpain inhibitor 1 (CPI-1) and calpain inhibitor 2 (CPI-2) each inhibited NOX- production and inducible iNOS expression in a concentration-dependent manner at 1 100 microM. TPCK and CPI-1 were about 10-fold more potent than TLCK and CPI-2, respectively. These data suggest that a chymotrypsin-like serine or cysteine proteinase is required for the LPS-inducible expression of the iNOS gene, perhaps by mechanisms involving activation of transcription factor NF-kappa B. Accordingly, a potent inhibitor of NF-kappa B activation whose action is attributed to inhibition of the chymotrypsin-like activity of the multicatalytic proteinase complex (MPC) was tested. Z-IE(O-t-Bu)A-Leucinal abolished NOX- production and inducible iNOS expression at 1 microM and showed over 50% inhibition at 10 nM. These observations indicate that inhibitors of MPC interfere with iNOS induction and provide strong evidence that MPC functions importantly in iNOS induction in macrophages. PMID- 7488012 TI - Dephosphorylation of a 30-kDa protein of fowl spermatozoa by the addition of myosin light chain kinase substrate peptide inhibits the flagellar motility. AB - Phosphorylation of demembranated fowl sperm proteins during incubation with [gamma-32P]ATP and various protein kinase substrate peptides at 30 degrees C was analyzed by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE). A marked difference in phosphorylation was observed in a 30 kDa protein. This protein was strongly phosphorylated after the addition of Kemptide, a cAMP dependent protein kinase (PKA) substrate peptide; Syntide 2, a calmodulin dependent protein kinase II substrate peptide; a protein kinase C (PKC) substrate peptide; as well as control samples but only slightly phosphorylated in the presence of a myosin light chain kinase (MLCK) substrate peptide. The motility of demembranated spermatozoa at 30 degrees C remained high in control samples and following the addition of Kemptide, Syntide 2 and PKC substrate peptide, but decreased markedly following the addition of MLCK substrate peptide. These results suggest that the 30 kDa protein is identified as a substrate for MLCK or a MLCK-like protein in fowl spermatozoa and that phosphorylation dephosphorylation of this protein is involved in the regulation of flagellar movement at 30 degrees C. PMID- 7488015 TI - Chain length-dependent Pb(II)-coordination in phytochelatins. AB - UV/visible and circular dichroism (CD) spectroscopy have been used to study the binding of Pb(II) to plant metal-sequestering peptides, phytochelatins (PCs), with the structure (gamma Glu-Cys)2Gly, (gamma Glu-Cys)3Gly and (gamma Glu Cys)4Gly. Saturation of the Pb(II)-induced charge-transfer bands indicated that both (gamma Glu-Cys)2Gly and (gamma Glu-Cys)3Gly bound one metal ion per peptide molecule. However, (gamma Glu-Cys)4Gly formed two distinct species with stoichiometries of one and two Pb(II) ions per peptide molecule, respectively. The optical spectra of Pb(II)1-(gamma Glu-Cys)4Gly were similar to those of Pb(II)1-(gamma Glu-Cys)3Gly, whereas the spectra of Pb(II)2-(gamma Glu-Cys)4Gly were similar to those of Pb(II)1-(gamma Glu-Cys)2Gly. Since cysteinyl thiolates are the likely ligands for Pb(II) in PCs, Pb(II) appears to form two-, three- and four-coordinate complexes with PCs depending on their chain length. Furthermore, Pb(II) may exhibit multiple coordination in longer chain PCs as indicated by the formation of two Pb(II)-binding species of (gamma Glu-Cys)4Gly. The transfer of Pb(II) from glutathione to PCs and from shorter chain to longer chain PCs is also demonstrated. PMID- 7488016 TI - Expression of the chemokine receptor BLR2/EBI1 is specifically transactivated by Epstein-Barr virus nuclear antigen 2. AB - In our attempt to identify chemokine receptors that are related to Burkitt's lymphoma receptor 1 (BLR1) and are expressed in activated lymphocytes we used RT PCR resulting in the isolation of a cDNA encoding a seven transmembrane receptor termed BLR2. The protein shows significant sequence similarities to the family of G-protein coupled chemokine receptors and turned out to be identical to the recently described receptor EBI1. Northern blot analysis revealed that BLR2 mRNA could be highly stimulated in mitogen- and anti-CD3-treated peripheral blood lymphocytes. BLR2-specific mRNA could be detected in all Epstein-Barr virus positive B cell lines. We show that transcription of the BLR2 gene could be specifically induced in Epstein-Barr virus negative BL 41 cells via estrogen mediated activation of Epstein-Barr virus nuclear antigen 2, a key regulator of viral and cellular genes in immortalized B cells. Our data suggest an involvement of BLR2 in the regulation of migration in activated lymphocytes and in viral pathogenesis. PMID- 7488017 TI - A novel sequence found at the 3' terminus of hepatitis C virus genome. AB - The 3' end region of positive-strand RNA-virus genomes is implicated in the initiation of genomic replication. We analyzed the extreme 3' end of the hepatitis C virus (HCV) genome by primer extension of the 5' end region of the antigenomic strand RNA found in infected liver. We discovered a novel sequence present downstream of the poly(U) stretch that was previously considered to be the 3' end structure of the HCV genome. The novel sequence was 98 nucleotides long and had no significant homology with any known sequences, viral or nonviral. The discovery of this novel tail on the HCV genome should contribute to the study of HCV replication. PMID- 7488018 TI - A novel method to purify recombinant vascular endothelial growth factor (VEGF121) expressed in yeast. AB - VEGF is a potent mitogen for vascular endothelial cells in vitro and acts as an angiogenic factor in vivo. VEGF121 differs from the other isoforms in that it lacks the heparin-binding domain. To study the potential differences in biological functions of the VEGF isoforms, we cloned and expressed VEGF121 in a yeast expression system. VEGF121 was secreted from the yeast cells as a homodimer with a molecular weight of 34-36 kDa. By taking advantage of the consecutive histidine residues present at position 11 and 12 in VEGF, a novel method of purification using Nickel affinity chromatography was developed. Since all the isoforms of VEGF have an identical amino terminal end, this method can be used to purify not only VEGF121 but also the other forms of VEGF. The level of expression achieved using this system was as high as 40 mg/L. The recombinant protein was biologically active in stimulating the in vitro proliferation of vascular endothelial cells and positively reacted to an antiserum made against recombinant VEGF165. PMID- 7488020 TI - The sugar composition and partial structure of the self-induced endogenous elicitor from potato. AB - The self-induced endogenous elicitor isolated from potato having molecular weight 9200 was degraded with trifluoroacetic acid to determine the sugar composition. The requisite composition of the elicitor was estimated to be a 2:3:4:7 mixture of galactose, rhamnose, arabinose, and galacturonic acid. Furthermore, the structure of oligogalacturonic acid moiety was found to consist mainly of alpha 1,4-pentagalacturonic acid by use of high pH anion exchange chromatography-pulsed amperometric detection. PMID- 7488019 TI - An alternatively spliced form of the human lysosome-associated membrane protein-2 gene is expressed in a tissue-specific manner. AB - We report the isolation of an alternatively spliced human lysosome-associated membrane protein-2 (h-lamp-2) transcript which is overexpressed in human muscle. The cloning of this transcript is an indication for the tissue-specific expression of lysosomal membrane proteins and implicates the possibility of multiple functions for the protein products of the h-lamp-2 gene, as well as other lysosome-associated membrane proteins. The new transcript, designated h lamp-2b, results from the alternative splicing of the last exon, exon 9, the alternative form of which is approximately 2800 bp in length. The resulting protein is identical in length to the previously reported h-lamp-2 protein, 410 amino acids including the leader peptide. This final exon, which encodes the last eleven amino acids of the luminal domain, the 24 amino acid transmembrane spanning region, and an eleven amino acid cytoplasmic tail, shows complete conservation of the Gly.Tyr.X.X lysosomal targeting signal with regard to its position relative to the transmembrane spanning region and the carboxy terminus of the protein. Immune electron microscopy studies verified localization of this alternative gene product to the lysosomal membrane. PMID- 7488021 TI - Expression of the type 1 and 2 steroid 5 alpha-reductases in human fetal tissues. AB - Androgens play a key role in human fetal development. Therefore, it is important to understand the distribution of the isozymes of 5 alpha-reductase, the enzyme that converts testosterone to the more potent androgen, dihydrotestosterone. The expression of the two isozymes of 5 alpha-reductase was studied in human fetal skin and fetal prostate by measuring the in vitro enzyme activity in crude preparations. Low levels of the type 1 5 alpha-reductase were found in fetal scalp and back skin. Studies with fetal prostate samples confirmed that the type 2 enzyme is expressed in levels similar to those found in adult tissues. These results provide the first evidence that both isozymes of 5 alpha-reductase are expressed during human fetal development. PMID- 7488023 TI - The mitochondrial DNA mutation ND6*14,484C associated with leber hereditary optic neuropathy, leads to deficiency of complex I of the respiratory chain. AB - The electron transfer activity of Complex I of the respiratory chain and Complex I-linked ATP synthesis were investigated in leukocytes of four males affected by Leber hereditary optic neuropathy and a mutation in the ND6 gene at nucleotide position 14,484 of mtDNA. The electron transfer activity in leukocytes of the patients was about 35% of that in control leukocytes, whereas the Complex I linked ATP synthesis showed a decrease of only about 20%. This demonstrates that all three mtDNA mutations that are clearly associated with Leber hereditary optic neuropathy result in deficiency of Complex I. However, the relationship between these mtDNA mutations, the function of Complex I and the phenotypic profile remains elusive. PMID- 7488022 TI - Only in the presence of immunophilins can cyclosporin and FK506 disrupt in vivo binding of calcineurin A to its autoinhibitory domain yet strengthen interaction between calcineurin A and B subunits. AB - The two immunosuppressants cyclosporin A (CsA) and FK506 exert their major therapeutic effect by inhibiting T-cell activation. It is believed that the drugs first bind to their cellular receptors, known as immunophilins, and then target the protein phosphatase calcineurin for inhibition. The catalytic activity of calcineurin is regulated by its autoinhibitory domain (AID) and by the calcium binding proteins calcineurin B (CnB) and calmodulin. We have used the yeast two hybrid system to show that AID, CnB and calmodulin can only bind to a truncated catalytic subunit of yeast calcineurin (i.e.,CnA1 delta), devoid of AID, but not to full-length CnA1. Both CsA and FK506 cause disruption of the CnA1 delta-AID interaction, whereas their presence permits CnA1 delta to bind more strongly to CnB. In contrast, the binding of CnA1 delta to calmodulin is unaffected by the immunosuppressants. Significantly, in the absence of its cognate cytosolic receptor, neither CsA nor FK506 inhibits or stimulates the CnA1 delta-AID, CnA1 delta-CnB interactions. These in vivo observations not only provide supportive evidence for the mechanism by which drug-receptor complexes could modulate calcineurin activity but also unveil the possibility of identifying novel immunophilin-independent calcineurin inhibitors which may disturb the association of CnA1 delta to AID. PMID- 7488025 TI - Depolarization of cell membrane is associated with an increase in ciliary beat frequency (CBF). AB - We hypothesize that activation of muscarinic cholinergic receptors depolarizes the cell membrane of the mammalian ciliated cells which in turn causes an increase of CBF. To test this hypothesis, a di-8-ANEPPS fluorescence photon counting and nonstationary heterodyne laser light scattering system was developed to measure cell membrane potential (psi) and CBF in cultured ovine tracheal ciliated cells simultaneously. Carbachol dose dependently depolarized the cell membrane with a corresponding stimulation of CBF. The carbachol induced depolarization of cell membrane and increases of CBF were inhibited by prior application of either atropine or verapamil or amiloride. These novel data suggest that depolarization of the cell membrane and the corresponding stimulation of CBF caused by the activation of muscarinic receptors of the mammalian ciliated cells are dependent on the influx of either extracellular Ca2+ or Na+. PMID- 7488024 TI - The role of the TATA box in the hormonal regulation of phosphoenolpyruvate carboxykinase gene expression. AB - In this study, we analyzed the role of the TATA box in the regulation of the phosphoenolpyruvate carboxykinase (PEPCK) gene expression by dexamethasone (DEX), retinoic acid (RA), glucagon (via cAMP) and insulin (INS). The PEPCK TATA box (TATTTAAA) was absolutely required for both basal promoter activity and hormone mediated transactivation. However, the relative induction of PEPCK gene expression by DEX, RA and cAMP, and its repression by INS, remained unaltered despite the substitution of the PEPCK TATA box with TATA elements from the herpes simplex virus-thymidine kinase gene, gene 33 or a consensus TATA box sequence, TATAAA. The results indicate that the TATA box serves a permissive, but not defining, function in the response of the PEPCK gene to hormones, and that this function can be equally facilitated by heterologous TATA box elements. PMID- 7488026 TI - ATP activates the potassium channel and enhances cytosolic Ca2+ release via a P2Y purinoceptor linked to pertussis toxin-insensitive G-protein in brain artery endothelial cells. AB - ATP produced whole-cell potassium currents in cultured endothelial cells of the bovine brain cortical arteries. P2 purinoceptor agonists evoked similar currents with the order of their potency: 2-methylthio ATP > ATP >> alpha, beta-methylene ATP > or = UTP > or = ADP >> AMP. ATP-evoked currents were inhibited by GDP beta S, but not by pertussis toxin (PTX). Furthermore, a phospholipase C (PLC) inhibitor, protein kinase C inhibitor, or cAMP-dependent protein kinase inhibitor had no effect on the currents. In addition to these effects, ATP enhanced intracellular free Ca2+ concentration ([Ca2+]i) in the presence and absence of extracellular Ca2+, and this [Ca2+]i increase was not inhibited by a PLC inhibitor. These results, thus, provide an indication that ATP activates the potassium channel and enhances [Ca2+]i via a P2Y purinoceptor linked to a PTX insensitive G-protein, which is not involved in a PLC-mediated signaling pathway. PMID- 7488027 TI - Structure and chromosomal localization of the human anti-mullerian hormone type II receptor gene. AB - Using the rat anti-mullerian hormone type II receptor (AMHRII) cDNA as a probe, two overlapping lambda phage clones containing the AMHRII gene were isolated from a human genomic library. Sequence analysis of the exons was performed and the exon/intron boundaries were determined. The coding region was found to consist of 11 exons, divided over 8 kb. The genomic structure resembles that of the related activin type II receptor gene. The AMHRII gene was mapped to human chromosome 12q12-q13. The results reported are essential for identification of AMHRII gene alterations in patients with persistent mullerian duct syndrome. PMID- 7488028 TI - Phosphatidylinositol 3-kinase is involved in TRE-dependent gene expression in response to arginine vasopressin. AB - We identified arginine vasopressin (AVP) as a potent activator of TPA-response element (TRE)-dependent gene expression in rat 3Y1 fibroblasts. In order to examine the involvement of phosphatidylinositol 3-kinase (PI3-kinase) in TRE mediated gene expression, we examined the effect of the overexpression of PI3 kinase mutant. The overexpression of p110 alpha, the catalytic subunit of PI3 kinase, enhances TRE-reporter expression in response to AVP. On the other hand, the overexpression of the p110-EcoS mutant, which contains the binding site for the regulatory p85 subunit but lacks the catalytic domain, results in decreased TRE reporter expression in response to AVP. These results suggest that PI3-kinase is involved in TRE-dependent gene expression in response to AVP. PMID- 7488029 TI - In vivo isoproterenol treatment leads to downregulation of the mRNA encoding the cAMP response element binding protein in the rat heart. AB - The expression of different myocardial regulatory proteins is altered in human heart failure, e.g., beta 1-adrenoceptors, G-proteins and others. Similar changes in rats after 4 days treatment with isoproterenol led to the hypothesis of the cAMP pathway involved in these changes. In different cell types cAMP-dependent transcriptional activation is mediated by the cAMP-response element binding protein (CREB) which was recently shown to be expressed and phosphorylated in the human heart. Here, by the reverse transcriptase-polymerase chain reaction two alternatively spliced isoforms of CREB mRNA were found to be expressed in rat ventricles. Both isoforms were down-regulated in the ventricles of rats treated in vivo with isoproterenol (2.4 mg/kg per day) for 4 days proposing a possible mechanism involved in expressional changes mentioned above. PMID- 7488030 TI - Expression, purification and functional characterisation of a Kunitz-type module from chicken type VI collagen. AB - The primary amino acid sequence of the carboxyl-terminal portion of the alpha 3 chain of chicken type VI collagen (K-VI) presents a 58-residue motif with a high degree of homology with members of the Kunitz serine-proteinase inhibitors family. This module was cloned, expressed in E. coli, purified and compared to the bovine pancreatic trypsin inhibitor (BPTI) in an inhibition profile assay of two serine proteases, trypsin and plasmin. We found that recombinant K-VI is not endowed with inhibitory activity but it slightly activates both plasmin and trypsin, differently from other members of the family. Moreover, the ability to inhibit the serine protease activity is also lacking in the intact type VI collagen molecule. PMID- 7488031 TI - Hyperphosphorylation of the cytoskeletal protein Tau by the MAP-kinase PK40erk2: regulation by prior phosphorylation with cAMP-dependent protein kinase A. AB - PK40erk2 is a MAP kinase which phosphorylates recombinant hTau40 up to 14 moles of phosphate/mole, markedly slowing its electrophoretic mobility. PK40erk2 acting on TAU is expected to cause the appearance of Alzheimer's disease-specific phosphoepitopes, detectable by specific antibodies. Maximal phosphorylation in vitro of hTau40 by PKAcat incorporates only 2-3 moles of phosphate/mole. Consequent, but smaller, reduction in electrophoretic mobility is seen, but not the formation of Alzheimer-specific or hyperphosphorylation-specific epitopes. Phosphorylation of hTau40 by PKAcat sharply reduces the number of phosphates that can now be introduced by PK40erk2 to 5-6 moles/mole, instead of the expected 11 moles/mole. Thus, prior phosphorylation by PKA, a non-proline-directed protein kinase, regulates the conformation of the protein substrate Tau so as to make some sites very much less accessible to phosphorylation by the proline-directed kinase, PK40erk2. PMID- 7488032 TI - The hydrophilic domain of phospholamban inhibits the Ca(2+)-ATPase--the importance of the method of assay. AB - The peptide MEKVQYLTRSAIRRASTIEMPQQAR-Cys representing residues 1-25 of phospholamban (PLN) decreases by 40% the maximal state rate of ATP hydrolysis by the Ca(2+)-ATPase of skeletal muscle sarcoplasmic reticulum (SR), measured at saturating concentrations of Ca2+. The pattern of Ca2+ uptake by SR vesicles in the presence of oxalate is complex, with an initial fast phase being followed by a lag phase and a second, slower phase of Ca2+ accumulation. PLN(1-25) reduces the rate of the slower phase of Ca2+ accumulation by 30%. However, if the level of accumulation of Ca2+ is measured after 2 min., the effect of PLN(1-25) is much less marked. It is concluded that PLN(1-25) inhibits the ATPase, but that the effects of this inhibition are not apparent under some assay conditions. PMID- 7488034 TI - c-Src phosphorylates epidermal growth factor receptor on tyrosine 845. AB - In the previous study [Sato et al. (1995) Biochem. Biophys. Res. Commun. 210, 844 851], we found that c-Src was associated with epidermal growth factor (EGF) receptor and activated upon EGF treatment in A431 cells. In the present study, we investigated the phosphorylation of EGF receptor by c-Src in the c-Src-EGF receptor complex. We have focused our attention to tyrosine residue 845 (Y845) of EGF receptor as a candidate for the phosphorylation site. A synthetic peptide containing Y845, named Y845 peptide, which corresponds to residue 837 to 856 of EGF receptor, was found to be phosphorylated by c-Src and used to provide the standard phosphopeptide. In addition to the autophosphorylated peptide of 25 kDa, a phosphopeptide of 7 kDa was detected in the cyanogen bromide-digested fragments of the c-Src-associated EGF receptor phosphorylated in vitro in an EGF-dependent manner. In phosphopeptide mapping, tryptic digest of the 7-kDa phosphopeptide was shown to co-migrate with that of the phosphorylated Y845 peptide. The 7-kDa phosphopeptide was found to be phosphorylated exclusively on tyrosine. These results suggest that c-Src can phosphorylate EGF receptor on Y845 in an EGF dependent manner. Furthermore, we confirmed that the same site of the c-Src associated EGF receptor was phosphorylated in EGF-treated A431 cells. PMID- 7488035 TI - Cloning, expression and functional role of XrpFI alpha and beta subunits in Xenopus laevis oocyte. AB - The cDNAs coding for the alpha and beta components of XrpFI, an activator of ribosomal protein genes transcription, were isolated. RNA analysis showed that transcripts of the beta genes are present at constant level throughout Xenopus development unlike the beta proteins which are undetectable in the early stages of embryogenesis when transcription is silent. On the contrary, alpha transcripts and proteins were found in all the embryo stages examined. The role of the XrpFI alpha and beta subunits in rp-gene transcription has been studied by mRNA microinjection into X.laevis oocytes. The presence of the beta subunit appears to be critical for the proper rp-gene promoter function. PMID- 7488033 TI - Dual effects of ATP on the potassium channel and intracellular Ca2+ release in smooth muscle cells of the bovine brain arteries. AB - ATP produced whole-cell potassium currents with a latency of 200 msec in cultured smooth muscle cells of bovine brain arteries. The currents were evoked via an ATP receptor linked to a pertussis toxin-insensitive G-protein, which is not involved in phospholipase C (PLC)-mediated signal transduction, and were not regulated by protein kinase A or C. In the cell-attached patches, ATP elicited single channel currents with two major classes of slope conductances and these currents were again induced within 100 msec after bath-application of ATP outside the patch pipette. In addition, ATP enhanced intracellular free Ca2+ concentration ([Ca2+]i) with no initiation time in the presence and absence of extracellular Ca2+, and this enhancement was not blocked by a PLC inhibitor. These results, thus, suggest that the activation of the potassium channel and the enhancement of [Ca2+]i induced by ATP may be caused by a direct action of the G-protein beta gamma subunits. PMID- 7488036 TI - Adenosine deaminase from human thyroid purification and some properties. AB - Gel filtration of human thyroid extract with Sephadex G-200 revealed two molecular forms of adenosine deaminase differing in their molecular sizes. The smaller form of adenosine deaminase was purified over 120-fold by precipitation of the protein impurities at pH5.6 and chromatography on DEAE-Sephadex A-50, Sephadex G-100 and adenosine-Sepharose. The purified enzyme was specific towards adenosine. The Michaelis constant was 5.2 X 10(-5) M. The optimum pH was about 7.0 and molecular weight 42000. PMID- 7488037 TI - Reciprocal regulation by cyclic nucleotides of the differentiation of rat osteoblast-like cells and mineralization of nodules. AB - The effects of cAMP and cGMP on the differentiation of osteoblast-like cells derived from rat calvariae and on the formation of bone in vitro were studied. Continuous culture of osteoblast-like cells in the presence of 8-bromo-cyclic AMP (8-Br-cAMP) resulted in the dose-related inhibition both of the synthesis of cellular alkaline phosphatase (ALPase), which is known as a marker of osteoblastic differentiation, and of the formation of mineralized nodules, which is a model of the formation of bone in vitro. By contrast, 8-bromo-cyclic GMP (8 Br-cGMP) promoted the synthesis of ALPase and the formation of mineralized nodules. Northern blot analysis revealed that these cyclic nucleotides modulated the steady-state levels of mRNAs for ALPase and osteocalcin, a bone-matrix protein that is specifically produced by osteoblast. The present results indicate that cAMP and cGMP act reciprocally to regulate osteoblastic differentiation and the subsequent formation of mineralized nodules. PMID- 7488038 TI - Peroxynitrite causes apoptosis in rat thymocytes. AB - Peroxynitrite is demonstrated to cause apoptosis in freshly harvested rat thymocytes suspended in phosphate buffer, as shown by the typical ladder pattern of DNA fragmentation. This evidence suggests a new function for peroxynitrite, already known to be a strong oxidant. Peroxynitrite can nick DNA, oxidize cellular thiols and initiate the apoptotic process responsible for genomic degradation. PMID- 7488039 TI - Cysteine 99 of endothelial nitric oxide synthase (NOS-III) is critical for tetrahydrobiopterin-dependent NOS-III stability and activity. AB - Tetrahydrobiopterin (BH4) is an essential cofactor for all three isoforms of nitric oxide synthase (NOS). However, its binding sites and functional roles remain elusive. Here, we demonstrated that cys-99 of human endothelial NOS (ecNOS) is critical for BH4 involvement in NOS catalytic activity and stability. Mutation of cys-99 to alanine in ecNOS resulted in loss of catalytic activity which could be restored to the level of wild type by adding a high concentration of exogenous BH4 to the crude extract. Purified C99A mutant was unstable and its maximal activity was only about 20% of the purified wild type activity. Comparison of BH4 concentration-dependent citrulline formation between C99A and the wild type revealed that the BH4 concentrations required for generating half maximal citrulline were 10-fold higher for C99A. Purified C99A had no detectable BH4 and had a reduced heme content when compared to the purified wild type, but retained the ability of forming CO-ferrous heme complex and had the same Km value for L-arginine (approximately 4 microM) as the wild type. These findings indicate that Cys-99 is critically involved in BH4 binding. Mutation of this residue leads to reduced affinity for BH4 and the resultant enzyme instability and irreversible heme loss. PMID- 7488040 TI - Didemnin B induces cell death by apoptosis: the fastest induction of apoptosis ever described. AB - Didemnin B, a cyclic N-methylated peptolide induces apoptosis in human HL-60 cells. When incubated with 1 microM didemnin B, unsynchronized HL-60 cultures undergo apoptosis to 100% within 140 minutes. Apoptosis has been assessed by the typical apoptotic morphology, the presence of double-stranded DNA fragments within the cytosol and the generation of DNA ladders. None of these characteristics of apoptosis are seen when HL-60 cells are pretreated with 1mM Zn2+ immediately before treatment with didemnin B. PMID- 7488041 TI - Dietary estrogenic isoflavones are potent inhibitors of beta-hydroxysteroid dehydrogenase of P. testosteronii. AB - The isoflavones daidzein, genistein, biochanin A and formononetin selectively inhibit the gamma-isozymes of mammalian alcohol dehydrogenase (ADH). Since gamma ADH is the only ADH isoform that catalyzes 3 beta-hydroxysteroid oxidation, it was conjectured that these isoflavones might also inhibit other enzymes involved in 3 beta-hydroxysteroid metabolism. P. testosteronii beta-hydroxysteroid dehydrogenase (beta-HSD) was used to evaluate this hypothesis. Indeed, all isoflavones that inhibit gamma-ADH were found to be potent inhibitors of beta HSD. Both the 3 beta- and 17 beta-HSD activities of the enzyme are inhibited. Kinetic analyses with pregnenolone (3-beta-OH) and testosterone (17-beta-OH) as substrates reveal that daidzein and genistein inhibit beta-HSD competitively with respect to the sterol substrates. Their Ki values are very similar and range from 0.013 to 0.02 microM. These results suggest that isoflavones may exert some of their biological effects by modulating activities of enzymes that metabolize steroids critical to hormonal and/or neuronal functions. PMID- 7488042 TI - Molecular cloning and organization of the coding region of the human norepinephrine transporter gene. AB - A lambda phage genomic library was screened with the digoxygenin labeled cDNA of the human norepinephrine transporter (hNET). Six overlapping lambda clones were analysed by restriction enzyme analysis and sequencing of the exon-intron boundaries. The coding region of the hNET gene was found to be encoded by 14 exons, spanning 45 kb from the start to the stop codon, disrupted by 13 introns. The organization of the gene is highly homologous to other known neurotransmitter transporter genes. However, the hNET gene differs from the other genes in that it has an additional exon encoding the C-terminus of the protein. The gene structure shows two large introns in the 5'-region and a cluster of 11 exons in the 3' region. All exon-intron junctions contain the gt/ag consensus splice site. Knowledge of the gene structure of the antidepressant-sensitive hNET should facilitate investigation of its potential role in psychiatric disorders. PMID- 7488043 TI - NMDA-dependent prostaglandin E2 release by human cultured astroglial cells is driven by nitric oxide. AB - The role of the L-arginine-NO pathway on the formation of PGE2 by human cultured astroglial cells incubated with NMDA has been investigated. Preincubation of T 67 astroglial cell line with NMDA (10-600 microM) produced a significant dose dependent increase of both nitrite (the breakdown product of NO), PGE2 and cGMP levels in cell supernatant. This effect was inhibited by coincubation of cells with L-NAME (20-300 microM), an inhibitor of NO synthase showing that the release of PGE2 subsequent to NMDA receptor stimulation was driven by NO. The release of PGE2 but not elevation of nitrite and cGMP levels was affected by indomethacin (10 microM), an inhibitor of cyclooxygenase. The inhibitory effect of L-NAME on PGE2 release by NMDA-pretreated astroglial cells was reverted by arachidonic acid, showing that the effect of NO on PGE2 release occurred at the cyclo oxygenase level. Thus, the present experiments demonstrate that the release of PGE2 by astroglial cells pretreated with NMDA is driven by activation of the L arginine-NO pathway, and this may be relevant in the pathophysiological mechanisms where glutamatergic neurotransmission is involved. PMID- 7488044 TI - Use of protease sensitivity to probe the conformations of newly synthesised mutant forms of mitochondrial aspartate aminotransferase. AB - Sensitivity to digestion with pronase has been used to show that the precursor form of mitochondrial aspartate aminotransferase, the form lacking the N-terminal presequence, that with a deletion of the first 9 residues and mutants of the mature enzyme in which residue Cys-166 is mutated to alanine or serine, all retain unfolded conformations after synthesis in a reticulocyte lysate. In the presence of lysed mitochondria the various forms of mitochondrial aspartate aminotransferase retained their susceptibilities to pronase in a way that mirrored the efficiencies with which they are imported into intact mitochondria. The results are interpreted as showing that the presequence of mitochondrial aspartate aminotransferase is not uniquely required for interaction with cytosolic factors required to maintain the newly synthesised protein in a form competent for interacting with, and being imported into, mitochondria. PMID- 7488045 TI - Reduction of mono(ADP-ribosyl)ation of histones in rat testis by gonadotropin testosterone system. AB - Mono(ADP-ribose) synthetase activity in the nuclear fraction of immature rat testis was investigated in reference to the effect of testosterone. When the nuclear fraction was incubated with [32P] NAD in the presence of nicotinamide, an inhibitor of poly (ADP-ribose) synthetase, some proteins with a molecular mass of 14.4-21.5 kDa were ADP-ribosylated. In these proteins, H2B and H3 histones were found by sequencing 20 amino acids in the N-terminus. Their incorporations of ADP ribose moiety were significantly decreased in immature rats at 4 hr after the subcutaneous injection of testosterone. Furthermore, the treatments with LH and FSH of the immature rats reduced mono(ADP-ribosyl)ations of H2B and H3 histones in a time-dependent manner. These results suggest that the testicular mono(ADP ribose) synthetase activity is under the control of gonadotropin-testosterone system and is possibly related to differentiation of the testis. PMID- 7488047 TI - Age-dependent changes of NO synthase activity in the rat brain. AB - We report that NO synthase activity, as expressed by citrulline and nitrite formation in brain homogenates, is decreased in 24-month old in comparison to 3 month old rats. In particular, a Ca(++)-dependent NO synthase activity was detected in homogenates obtained from cortical, hippocampal, cerebellar and lower brain stem slices from both 3- and 24 month-old rats. The amount of citrulline generated from L-arginine was significantly decreased in the hippocampus and lower brain stem by 40 and 48%, respectively. No changes were observed in NO synthase activity in cortical and cerebellar homogenates. Thus, the L-arginine-NO pathway seems to be impaired in selected areas of rat brain and this may contribute to the understanding of pathophysiological mechanisms underlying age related cerebral disorders. PMID- 7488046 TI - Multiple types of ryanodine receptor/Ca2+ release channels are expressed in vascular smooth muscle. AB - Functional studies on vascular smooth muscle suggest the presence of ryanodine receptors (RyRs) with differing properties. In an attempt to understand such differences we investigated, using reverse transcription-polymerase chain reaction (RT-PCR), the possibility that smooth muscle cells express multiple types of RyRs. RNA was extracted from rat aorta, superior and small mesenteric arterial vessels, purified aortic smooth muscle media, or cultured aortic smooth muscle cells for cDNA synthesis. The cDNAs encoding RyR1, RyR2, and RyR3 were amplified using PCR primers based on sequences close to the 3' coding region of the RyR genes and PCR products verified by restriction endonuclease analysis. All three members of the RyR gene family were found to be present in vascular smooth muscle. This finding of multiple types of RyRs expressed in the same cell type indicates a complex mechanism of RyR Ca2+ channel regulation involving the formation of homo- and/or heterotetrameric complexes. PMID- 7488049 TI - Novel nonsense proteolipid protein gene mutation as a cause of X-linked spastic paraplegia in twin males. AB - We report a third mutation of the proteolipid protein gene in male Japanese patients with X-linked spastic paraplegia. Although the proteolipid protein gene encodes two myelin proteins, proteolipid protein and DM 20, our W144X mutation resides in the latter part of exon 3 (exon 3B), which is spliced out in DM 20. This mutation may reserve the function of DM 20. Findings in our patients support that this form of spastic paraplesia is allelic to Pelizaeus-Merzbacher disease and that the mild clinical phenotype of this disorder may be related to a mutation within exon 3B of the PLP gene. PMID- 7488048 TI - Such hydrophobic peptides as dansylated mastoparan can elevate the fertilization membrane of sea urchin eggs. AB - Melittin is known to be a major hydrophobic peptide component in honeybee venom that can cause as much elevation of fertilization membrane of sea urchin eggs as normal fertilization. The action of melittin has been thought to be closely related with its ability to facilitate the phospholipase A2 activity on the eggs. However, another peptide "mastoparan" from wasp venom was not found here to cause any elevation of the membrane, although it can activate the enzyme as well as melittin. On the other hand, mastoparan was found to get the membrane-elevating activity only when its amino groups were modified with hydrophobic substituents. N epsilon-Substituted mastoparan with a dansyl group in Lys11 residue was most effective among the analogs examined here. Our findings indicate that the facilitation of phospholipase by the peptides have little relation with the membrane generation. Such hydrophobic moiety as the dansyl group in the peptides must cause the cortical reaction on the eggs in cooperation with peptide moiety. The dansylated peptide will be a useful tool to induce the artificial fertilization of sea urchin eggs. PMID- 7488050 TI - Direct measurement of nitric oxide production in platelets: relationship with cytosolic Ca2+ concentration. AB - NO production in platelets has been followed by electrochemical detection. It was undetectable in unstimulated platelets and in thrombin or ADP-stimulated platelets, but dose-dependently stimulated by collagen. A production of 5 10(-19) mol/platelet was reached with 9 micrograms collagen. In collagen-stimulated platelets, preincubation with 1 mM L-Arg, D-Arg or L-NMMA increased by 77%, left unchanged or decreased by 63% NO production, respectively. NO production did not parallel cytosolic Ca2+ changes, although it decreased in low Ca2+ medium or when Ca2+ transients were attenuated by intracellular Ca2+ buffer. These results confirm that human platelets can generate NO. They demonstrate that cytosolic [Ca2+], although participating in the regulation of its synthesis, is not the messenger for NO synthase activation. Platelet NO production could become functionally important when collagen fibrils of the sub-endothelium are accessible. PMID- 7488051 TI - Antisense oligonucleotide inhibits the expression of procollagen type III gene in mouse fibroblasts. AB - The effects of transfection of phosphorothioate antisense oligonucleotides (ASOs) into BALB/c 3T3 mouse fibroblasts on procollagen type III production were examined. Immunohistochemistry and Western blots showed that the expression of procollagen type III decreased. Semi-quantitative reverse transcription polymerase chain reaction revealed that the expression of procollagen type III mRNA was inhibited. ASOs were effective for inhibiting procollagen type III production at the gene level, although the effect was transient. PMID- 7488053 TI - Purification and characterization of a novel protein activator of Ca2+/calmodulin dependent protein kinase I. AB - A protein activator of Ca2+/calmodulin (CaM)-dependent protein kinase I was purified from rat brain. The activator was retained on a CaM-Sepharose column in the presence of Ca2+ and kinase assay of renatured gel revealed the 64 kDa molecule in the purified activator fraction to be autophosphorylated and to phosphorylate recombinant CaM kinase I in the presence of Ca2+/calmodulin. These results suggest that this activator of CaM kinase I is also a CaM-dependent protein kinase. Phosphorylation of CaM kinase I by the activator resulted in drastic potentiation of its CaM-dependent activity. Furthermore, kinetic analyses demonstrated that the activation decreases the Km values of CaM kinase I for both ATP and syntide-2 without a change in Vmax values. Considering the quite low enzymatic activity of recombinant CaM kinase I without activation, the 64 kDa species might be essential for CaM kinase I function in vivo. PMID- 7488052 TI - Evidence of histidine coordination to the catalytic ferrous ion in the ring cleaving 2,2',3-trihydroxybiphenyl dioxygenase from the dibenzofuran-degrading bacterium Sphingomonas sp. strain RW1. AB - The 1H NMR spectra of an aromatic ring-cleaving extradiol dioxygenase, 2,2',3 trihydroxybiphenyl dioxygenase of the dibenzofuran-degrading bacterium Sphingomonas sp. strain RW1, are reported. In the catalytically active reduced form of the monomeric enzyme (MW = 32 kDa), three broad strongly downfield shifted signals were observed, two of which disappeared in D2O solution. Their shifts and linewidths are consistent with ring NH and meta-like protons of coordinated histidines. These signals show strong sensitivity to the presence of the substrate. The oxidized form of the enzyme shows no hyperfine shifted signals. It is suggested that the high spin Fe(II) ion present in the active form of the enzyme is coordinated by at least two histidines. This is the first report of hyperfine shifted NMR signals being detected for an extradiol dioxygenase. PMID- 7488055 TI - CTF/NF-1 binds the stage selector element of the human gamma-globin gene promoter. AB - The human gamma-globin gene undergoes to a transcriptional switch during development, being expressed in the fetus and silenced at birth. It has been proposed that the sequence centered at -50 of the gamma-globin promoter contains a stage selector element (SSE) involved in the gamma- to beta-globin switching. The presence of the SSE confers preferential functional interactions of the promoter with the locus control region (LCR) in the fetal liver. It has been shown that two factors bind the gamma-globin SSE: Sp1 and the stage selector protein (SSP). Here we show that also the ubiquitous transcription factor CTF/NF 1 binds to the -53 to -35 element of the human gamma-globin promoter and its specificity is not influenced by flanking sequences. PMID- 7488054 TI - Adrenomedullin suppresses mitogenesis in rat mesangial cells via cAMP pathway. AB - Adrenomedullin (ADM) is a vasoactive peptide that was recently localized in renal glomeruli. In the present study we explored whether ADM stimulates cAMP system in glomerular mesangial cells (MC) and whether it can via "negative-crosstalk" inhibit the mitogen-activated protein kinase (MAPK) and thus suppress proliferation of MC. We found that ADM elicited accumulation of cAMP and in situ activation of protein kinase A (PKA) in cultured MC. Addition of 1 nM ADM to incubation media inhibited the proliferation in both quiescent MC and cells maximally stimulated by PDGF and also decreased the activation of MAPK induced by PDGF. These results indicate that ADM can suppress MC mitogenesis and suggest that it may function as an endogenous paracrine supressor of MC proliferation. PMID- 7488056 TI - Characterization of human prodynorphin gene transcripts. AB - Prodynorphin gene transcripts have been characterized in human tissues with cRNA probes covering parts of the non-coding exon 1 and the main coding exon 4 using Northern blot and RNase protection experiments. A 2.8-kb signal was observed in human brain RNA with both the exon 1 and exon 4 probes. An RNase protection assay with the exon 1 probe, performed to map the 5' end of this transcript, produced protected fragments in the range of 0.11 to 0.15 kb indicating that exon 1 is 1.2 kb shorter than previously proposed. 5'-RACE-PCR and sequencing of amplified cDNA fragments confirmed this assignment. In adrenal gland, testis and the human small cell lung carcinoma cell line, U1690, several prodynorphin-like mRNAs structurally different from the brain prodynorphin mRNA species were observed. PMID- 7488058 TI - In vivo degradation of human fibrinogen A alpha: detection of cleavage sites and release of antithrombotic peptides. AB - Several degradation products of fibrinogen have been shown to possess regulatory functions. Using peptide extracts from human blood filtrate, a large number of fibrinogen A alpha fragments was identified. These fragments are generated at known plasmin attack sites and at several novel cleavage sites especially at hydrophobic and basic amino acid residues. One fragment containing the cell attachment site (RGD sequence) of fibrinogen A alpha efficiently inhibits fibrinogen binding and platelet aggregation (IC50:20-50 microM) in vitro. We conclude that in vivo degradation of fibrinogen A alpha results in generation of endogenous antithrombotic peptides with local importance in fibrinolysis and platelet aggregation. PMID- 7488059 TI - Ca(2+)-induced Ca2+ oscillations in parathyroid cells. AB - Parathyroid cells from patients with hyperparathyroidism were first cultured in a growth cocktail supplemented version of Dulbecco's MEM/F12 medium and then used for measurements of the cytoplasmic Ca2+ concentration ([Ca2+]i). Elevation of extracellular Ca2+ from 0.5 to 1.5-3 mM often triggered rhythmic oscillations of [Ca2+]i. Whereas the amplitude of the oscillations remained constant their frequency (0.4-2.6 /min) increased with the Ca2+ concentration and there was sometimes transformation into sustained increase of [Ca2+]i at the highest extracellular Ca2+ concentration. The oscillations were abolished by the inorganic Ca2+ channel blocker Ce3+ and by K+ depolarisation, which lowered [Ca2+]i, as well as by thapsigargin, an inhibitor of the intracellular Ca(2+) pumping ATPase. The data indicate that extracellular Ca2+ can trigger [Ca2+]i oscillations in parathyroid cells, dependent on mobilisation of the ion from intracellular Ca2+ stores and on influx through non-voltage dependent channels. PMID- 7488057 TI - Down-regulation of c-Myc and Max genes is associated to inhibition of protein phosphatase 2A in K562 human leukemia cells. AB - Treatment of the human myeloid leukemia K562 cells with the protein phosphatase inhibitors okadaic acid or calyculin A resulted in down-regulation of both c-myc and max genes at the mRNA and protein levels. The extent of the down-regulation was similar for both genes and was dependent on the dose and on the treatment time. Interestingly, c-myc and max down-regulation was concomitant with apoptosis induced by okadaic acid and calyculin A in K562 cells. The expression of c-myc and max returned to control levels after the removal of okadaic acid from the media, although apoptosis was irreversible. These effects were observed at okadaic acid concentrations (15 nM) that inhibited the activity of protein phosphatase type 2A but not of phosphatase type 1. We conclude that the inhibition of protein phosphatase 2A is associated to decreased levels of c Myc/Max heterodimers in K562 cells. PMID- 7488060 TI - Characterization of the human nm23-H2 promoter region and localization of the microsatellite D17S396. AB - The transcription of human nm23 genes (nm23-H1, nm23-H2) is involved in suppression of tumor metastasis or tumor progression. Therefore the characterization of transcriptional regulatory mechanisms for both nm23 genes is very important. In this study we have isolated and analyzed the 5'-flanking region of the human nm23-H2 gene and estimated the distance to 4 kb between nm23 H2 and nm23-H1 genes. We localized the known microsatellite D17S396 within this region. Furthermore the identification of possible binding sites for MYC proteins and additionally the NM23-H2 protein itself (the transcription factor PuF for c myc gene activation) is of importance with respect to possible para- and autoregulatory interactions. A comparison of the promoter sequences of both human nm23 genes revealed no significant sequence homology. PMID- 7488061 TI - Properties of the Kv1.1 rat brain potassium channels expressed in mammalian cells: temperature effects. AB - We studied the temperature dependence of the currents mediated by potassium channels Kv1.1 cloned from rat brain expressed by permanent transfection in the human embryonic kidney cell line 292er. Voltage-gated outward currents, with a reversal potential near to the Nernst potential for potassium, were elicited by depolarising pulses. The currents activate fast and do not show inactivation during 50-ms test pulses. Increasing temperature decreased the amplitude of the currents, slowed the kinetics of activation and deactivation and shifted the activation curve to more negative potentials. We conclude that in Kv1.1 channels the closed states are associated with more ordered structure of the channel protein than the open state. The voltage-dependence and the kinetic properties are similar to those expressed in frog oocytes injected with cRNA coding the same Kv1.1. PMID- 7488062 TI - Interferon-alpha primes macrophages for lipopolysaccharide-induced apoptosis. AB - Apoptosis plays an important role in generating and maintaining an effective immune system. Many pathogens can perturb the homeostasis of the immune system by either inducing or suppressing cell death of immune cells. Using bovine macrophages as a model, we found that interferon-alpha, one of the host's responses to viral infection, can prime macrophages for activation-induced apoptosis. Exposure of bovine bone-marrow-derived macrophages to interferon-alpha and subsequent activation with lipopolysaccharide led to a strong downregulation of the macrophages' nitric oxide production when compared to lipopolysaccharide stimulation alone. We could show that this was due to induction of apoptosis after activation of the cells. Herpesvirus-induced type I interferon also primed bovine macrophages for lipopolysaccharide-induced apoptosis. Our studies describe how in a novel pathway an antiviral immune response could contribute to pathological sequelae of viral diseases. PMID- 7488063 TI - Efficient cleavage of p220 by poliovirus 2Apro expression in mammalian cells: effects on vaccinia virus. AB - Poliovirus protease 2A cleaves p220, a component of initiation factor eIF-4F. Polyclonal antibodies that recognize p220 and the cleaved products from different species have been raised. Transfection of several cell lines with poliovirus 2Apro cloned in different plasmids leads to efficient cleavage of p220 upon infection with VT7, a recombinant vaccinia virus that expresses the T7 RNA polymerase. Under these conditions vaccinia virus protein synthesis is severely inhibited, while expression of poliovirus protein 2C from a similar plasmid has no effect. These results show by the first time the effects of p220 cleavage on vaccinia virus translation in the infected cells. PMID- 7488064 TI - Chromaffin granule aspartic proteinase processes recombinant proopiomelanocortin (POMC). AB - Our search for proteases responsible for proenkephalin (PE) processing in adrenal medulla led to the isolation of a 70 kDa aspartic proteinase that cleaves PE between the basic residues of the Lys-Arg processing site (1). Studies in pituitary have also identified a similar aspartic proteinase that processes POMC (2,3). To compare the chromaffin granule (CG) 70 kDa aspartic proteinase with that in pituitary, processing of recombinant POMC by the CG enzyme was examined. POMC was expressed in the T7 expression system in E. coli, and purified to homogeneity. The CG 70 kDa aspartic proteinase converted POMC to 27 and 22 kDa bands that were detected by anti-N-POMC immunoblots, and to 26, 22, and 14 kDa bands that were immunoreactive with anti-beta-lipotropin. POMC products represented by these bands indicate appropriate POMC processing by the CG 70 kDa aspartic proteinase. These results, combined with the similar biochemical properties of these two enzymes, suggest that the CG 70 kDa aspartic proteinase resembles the POMC-converting enzyme (PCE), an aspartic proteinase in pituitary (2,3). PMID- 7488065 TI - pH dependent alterations of monoepoxides and monochlorohydrins of linoleic acid, and their existence in vivo. AB - Some monoepoxides of linoleic acid (LA) were converted to monochlorohydrins in low-pH solutions containing chloride ions (Cl-). Conversely, monochlorohydrins of LA were converted to monoepoxides in high-pH solutions. We attempted to determine whether these monochlorohydrins and monoepoxides were produced from LA by the cytochrome-c-H2O2-and/or myeloperoxidase-H2O2-system. The existence of monoepoxides and monochlorohydrins of LA in leukocytes was confirmed by high performance liquid chromatography (HPLC). Furthermore, leukotoxin in human leukemia cells (THP-1) was stained immunohistochemically by a monoclonal anti leukotoxin antibody. PMID- 7488066 TI - Mitochondrial DNA minicircles, lacking replication origins, exist in the cardiac muscle of a young normal subject. AB - This is the first report that mitochondrial (mt) DNA 'minicircles,' lacking replication origins, exist in the heart mtDNA of a young normal subject. A total detection system using 180 kinds of PCR primer pairs was recently devised to detect all possible deletions in mtDNA. To assess the reliability of the system, template mtDNAs were prepared from the heart muscle by three different methods including DNAase digestion. The methods for mtDNA preparation and the nuclear DNA contaminants were demonstrated to have few influences on the analysis. With not less than 95% accuracy, 52 types of deletions including 36 types of minicircles were detected in the subject's mtDNA. These results will give an insight to the mechanism of mtDNA deletion and to the relevance of the deletion with normal aging. PMID- 7488067 TI - High level production and localization of bovine rod cGMP-gated cation channel subunit 1 in baculovirus-infected insect cells and Saccharomyces cerevisiae. AB - The cGMP-gated cation channel is responsible for the last step in the vertebrate phototransduction cascade which couples light activation of rhodopsin to a change in membrane permeability. Two different expression systems, baculovirus-infected Spodoptera frugiperda (Sf9) insect cells and Saccharomyces cerevisiae, were used for the overproduction of the cGMP-gated cation channel subunit 1 of bovine rod cells. Presence of recombinant channel protein was monitored by SDS-PAGE and Western-blot analysis. Through immunogold labeling, the heterologously expressed cation channel was found to be localized predominantly in the inner compartments of the infected insect cells and in the vacuole of recombinant yeast cells. PMID- 7488068 TI - Biocatalytic oxidation of polycyclic aromatic hydrocarbons by hemoglobin and hydrogen peroxide. AB - Hemoglobin is able to oxidize polycyclic aromatic hydrocarbons, PAH's, in presence of hydrogen peroxide. Among 12 aromatic compounds tested, six were oxidized; anthracene, carbazole, dibenzothiophene, fluorene, 9-hexylanthracene and pyrene. The products were identified as aromatic ketones and sulfoxides. Effect of organic solvent concentration and hemoglobin stability were determined. PMID- 7488069 TI - Thrombin receptor polymorphism in Chinese hamster. AB - We report the existence of a new Chinese hamster thrombin receptor allele, characterized by an in-frame insertion of three nucleotides at position 250 of the published sequence. As a consequence, an additional proline is inserted into a proline-rich region of the extracellular amino-terminal domain of the receptor. A corresponding proline at this position is also found in the rat thrombin receptor. A silent base-pair change is found in the cytoplasmic tail of the receptor gene. Single-strand conformation polymorphism and sequence analysis indicate this new receptor allele is present in several cell lines derived from different individual Chinese hamsters. Embryonic CHEF IIC9 cells and primary culture cells are homozygous for this new allele. In contrast, the CCL39 lung fibroblast cell line is heterozygous for both the new and old alleles. Both alleles are transcribed into mRNA and code for functional receptors. Given the allelic distribution and sequence alignment with thrombin receptors from other species, we propose that the new sequence represents the actual predominant allele in Chinese hamster. PMID- 7488070 TI - Endothelin induced collagen remodeling in experimental pulmonary hypertension. AB - To investigate pathophysiological roles of endothelin-1 (ET-1) in collagen remodeling in pulmonary hypertension, we measured: (a) mRNA expression, concentration, localization of ET-1; (b) changes in types and content of collagen in the lung; (c) and confirmed direct effects of ET-1 on type V collagen metabolism in vascular smooth muscle cells. Monocrotaline-treated rats showed pulmonary hypertension with medial hypertrophy and perivascular fibrosis of pulmonary arteries. At the progressive stage of pulmonary hypertension, both ET-1 levels and its mRNA expression in the lung increased. Total collagen in the lung rose markedly with a higher rate of increase in type V collagen. ET-1, which exists in vascular smooth muscle cells, other perivascular cells and endothelium, stimulated type V collagen production. Our results suggest that local production of ET-1 in the lung contributes to progression of pulmonary hypertension through changes in phenotypes and content of collagen. PMID- 7488071 TI - DNA end-joining in extracts from human cells. AB - DNA end-joining is a central feature of several DNA recombination processes. A DNA end-joining activity present in extracts prepared from cells of the human SupT1 lymphocyte cell line was characterised. Joining of blunt ends and ends having complementary single-strand extensions (SSEs) were precise with no insertion or deletion of substrate base pairs. DNA sequencing analysis showed that molecules having non complementary ends of the same polarity, or molecules having one blunt end and one end with a SSE, were joined without loss of nucleotide sequences in the double-stranded region of the substrate molecule. The joining patterns observed have several features that are consistent with DNA end joining activities previously observed in vitro in extracts from Xenopus eggs and in vivo in mammalian cells and yeast. PMID- 7488072 TI - A novel Cl(-)-pump: intracellular regulation of transport activity. AB - Utilizing a purified basolateral plasma membrane vesicle preparation containing Cl(-)-ATPase, it was demonstrated that ATP, and its subsequent hydrolysis, stimulated both intravesicular Cl- accumulation and intravesicular negativity with almost identical kinetics. Mg2+ also stimulated both pump transport characteristics with an optimal concentration of 3 mM. Similarly, the pH optimum for both pump transport expressions was 7.8. Orthovanadate inhibition of both pump transport characteristics was directly related to its concentration. These results suggested that the active Cl- transport mechanism is electrogenic and is driven by a Cl(-)-stimulated ATPase. PMID- 7488073 TI - The neuroendocrine protein 7B2 contains unusually potent transcriptional activating sequences. AB - The expression of the 7B2 protein, secreted from a variety of neural and endocrine tissues, increases dramatically in specific neuroendocrine tumors. We have recently shown that human 7B2 can act as a molecular chaperone in the deaggregation of proteins in vitro. In order to identify polypeptides which might bind 7B2 in vivo, the yeast two-hybrid system was employed. Surprisingly, mere covalent linkage of 7B2 to the DNA-binding domains of two yeast transcription activators, Ace1 and Gal4, activates transcription from the ACE1 and GAL4 operon. 7B2's ability to activate nuclear transcription surpasses that of Ace1 and compares favourably with the strong activation domain of the tumor suppressor protein, p53. Our results suggest that 7B2 must possess an activating sequence, a domain which defines all transcriptional activator proteins. Like the acidic activation domains of some transcriptional activators, 7B2 also binds the yeast TATA-box binding protein, an essential polypeptide in the basic transcription machinery. Deletion analysis of the gene encoding 7B2 reveals two independent transcriptional activating sequences in the 185 amino acid protein. It is therefore conceivable that 7B2 not only has a functional role in the secretory pathway but also in the nucleus. Moreover, these findings raise an intriguing question regarding the activation domains of 7B2 and their possible link to 7B2's oncogenic potential. PMID- 7488074 TI - Differential activation of PKC isozymes by 14-3-3 zeta protein. AB - 14-3-3 proteins are ubiquitous in eukaryotes associated with many fundamental functions in signal transduction pathways and cell cycle regulation. Protein kinase C comprises a large family of serine/threonine protein kinases that are involved in cell growth and differentiation. Different protein kinase C isozymes have distinct roles in signal transduction pathways; protein kinase C epsilon is of particular interest because its overexpression leads to oncogenic transformation. The 14-3-3 protein has been reported to regulate the activity of protein kinase C, although the nature of its effect is equivocal. In this study we report the differential activation of various protein kinase C isoforms by 14 3-3 zeta protein. The classical isozymes show approximately a twofold activation, protein kinase C delta shows no significant increase in activity, whereas protein kinase C epsilon, another novel isozyme, is highly activated. This activation shows strong positive cooperativity with a Hill coefficient of 6.1 +/- 0.2. PMID- 7488075 TI - Essential fatty acid metabolism in cultured rat cardiomyocytes in response to either N-6 or N-3 fatty acid supplementation. AB - In this study we demonstrate that cultured rat cardiomyocytes possess the capacity to desaturate/elongate essential fatty acids (EFAs). Alpha-linolenic acid (ALA) conversion to higher metabolites was greater than linoleic acid (LA) conversion, according to the higher affinity of the delta-6-desaturase enzyme for the n-3 than for the n-6 EFAs. Gamma-linolenic acid (GLA) supplementation to the culture medium had no influence on LA conversion; but the addition of eicosapentaenoic (EPA) and docosahexaenoic (DHA) acids significantly decreased the formation of interconversion products from LA. The conversion of ALA to higher metabolites was greatly affected by GLA; EPA had no effect on ALA conversion, while DHA significantly inhibited it. Both GLA (converted mostly to dihomo-gamma-linolenic acid) and EPA can be removed from phospholipids and addressed to prostanoid biosynthesis, so avoiding their potential accumulation and the inhibition of their own production. Our data clearly indicate that supplementation of the culture medium with either n-6 or n-3 fatty acids can cause reduced levels of the other series of fatty acids. This effect may be undesirable, since both n-6 and n-3 fatty acids are important in the prevention of coronary diseases. PMID- 7488076 TI - Macrophage-stimulating protein activates Ras by both activation and translocation of SOS nucleotide exchange factor. AB - Macrophage-stimulating protein (MSP) is a chemotactic factor that activates the receptor tyrosine kinase RON. The involvement of Ras in MSP-induced signal transduction was investigated. Here we demonstrate that, in RON-transfected MDCK cells, an active GTP-bound form of Ras was rapidly accumulated by MSP treatment and the Ras-guanine nucleotide exchange activity in SOS immunoprecipitates was concomitantly increased. GAP activity was not changed under the same conditions used. Furthermore, the SH2 domain of adaptor protein GRB2, but not Shc, associated with the activated RON-beta chain, and GRB2-SOS complexes translocated from the cytosol to the membrane upon MSP treatment. These results strongly suggest that MSP activates Ras through RON, and that MSP-induced activation of Ras might be controlled by both the enhancement of catalytic exchange activity of SOS and its translocation to the membrane where its target Ras is localized. PMID- 7488077 TI - The Claviceps purpurea gene encoding dimethylallyltryptophan synthase, the committed step for ergot alkaloid biosynthesis. AB - The first pathway-specific step of ergot alkaloid biosynthesis in the fungus, Claviceps purpurea, is catalyzed by the prenyltransferase, 4-(gamma,gamma dimethylallyl)tryptophan synthase. Partial sequence information was obtained for the purified enzyme and a degenerate oligonucleotide mixture was used to identify and amplify segments of the gene, dmaW. The complete gene and near-full-length cDNA were cloned and sequenced. The cDNA was cloned in a yeast expression vector in sense and antisense orientations relative to the inducible GAL1 promoter. Extracts of yeast transformants with the sense constructs, but not antisense constructs or cloning vector, catalyzed production of 4-(gamma,gamma dimethylallyl)tryptophan. The sequence of dmaW and its cDNA indicated that it encoded a 455 amino acid polypeptide with a predicted molecular mass of 51,824 Da and a putative prenyl diphosphate binding motif. PMID- 7488078 TI - A brain-specific G protein gamma subunit. AB - Two different cDNAs for G protein gamma subunits have been isolated from mouse brain. One encodes a novel gamma subunit, gamma 4, the expression of which is detected only in brain. A fragment of this cDNA had been isolated previously. The other cDNA encodes gamma 3, a subunit type previously isolated from bovine brain. The primary structure of the gamma 3 subunit is conserved completely across species indicating that the diversity in the structure of the gamma subunits is of functional consequence. Moreover, gamma 2, gamma 3 and gamma 4, which are predominantly expressed in brain, are more homologous to each other than other gamma subunits, indicating that the G protein gamma subunits belong to distinct subfamilies similar to the alpha subunits. PMID- 7488079 TI - Regulation of rhodopsin phosphorylation by a family of neuronal calcium sensors. AB - Recoverin is a calcium sensor that regulates rhodopsin phosphorylation in a calcium-dependent manner. Cloning experiments indicate the presence of a numerous gene family, called the NCS family, encoding recoverin-like proteins expressed predominantly in neurons. Here, we report the cloning of three novel NCS genes, and demonstrate that at least six distinct members of the NCS family (including recoverin, S-modulin, vilip 1, NCS-1, Ce-NCS-1, and Ce-NCS-2) specifically inhibit rhodopsin phosphorylation. The presence of species homologues within the NCS family suggests that this function might be shared by at least 12 (out of 18) NCS proteins. Recent studies indicate that recoverin inhibits rhodopsin phosphorylation by directly regulating rhodopsin kinase, a G protein coupled receptor kinase (GRK). Since several NCS proteins are found in neurons throughout the entire nervous system, they may regulate other members of the GRK family. Together, our data suggest a general role for NCS proteins in the regulation of calcium-dependent phosphorylation in the nervous system. PMID- 7488080 TI - Adaptation to phosphate depletion in opossum kidney cells. AB - Dietary phosphate restriction produces an adaptive increase in renal tubular Na/Pi cotransport. A similar adaptation occurs during phosphate depletion in opossum kidney cells, a continuous line of cultured renal epithelial cells. We investigated the cellular changes associated with adaptation to phosphate depletion in OK cells, in isolation from the complex systemic changes that occur with in vivo phosphate restriction. Phosphate depletion for up to 24 hours was associated with increases in Na/Pi cotransport activity, Na/Pi cotransporter mRNA, and Na/Pi cotransporter protein. Moreover, the increases in Na/Pi cotransport, and Na/Pi cotransporter mRNA and protein occurred at physiologically relevant degrees of phosphate restriction. The experimental results suggest that increases in Na/Pi cotransporter mRNA and protein may mediate the increase in Na/Pi cotransport activity in OK cells during phosphate depletion. PMID- 7488082 TI - L-band ESR spectra of copper(II) complexes with CuN4 configurations. AB - L-band ESR spectrometry has been developed for detecting free radicals or paramagnetic metal ions in intact animals or biological samples which contain much water. Several studies by L-band ESR on free radicals have been reported, but few on paramagnetic elements such as copper(II) or iron(III). We measured L band ESR spectra of Cu(II) for the first time to investigate the coordination structure around the metal ion in aqueous solution. In X- and L-band ESR, both spectral patterns and parameters have been found to reflect the coordination structure of Cu(II)-polyamine complexes depending on the structures such as square-planar, axially coordinate square-planar, tetrahedral and distorted square planar (Cu(II) ion is displaced from CuN4 plane). In addition, in L-band ESR, the coordination structures of Cu(II) complexes relate with spectral pattern, ESR parameter (LA-value), stability constants and halfwave potentials in terms of the electron density on Cu(II). Thus, L-band ESR spectrometry was indicated to be useful to know the coordination structure of Cu(II) proteins or enzymes in aqueous solution. PMID- 7488081 TI - Overexpression of the obese gene in the genetically obese JCR:LA-corpulent rat. AB - Expression of the obese (ob) gene in JCR:LA-cp rats was examined. A 360 bp fragment of the conserved region of the gene was obtained by RT-PCR using total RNA isolated from adipose tissues of Sprague-Dawley (SD), JCR:LA-cp obese and lean rats. The three gene fragments were sequenced and shown to be identical. They were over 90% identical to the mouse ob gene sequence. The amplified fragments encode for 120 amino acids and have a glutamine residue at position +49. The gene was shown to be expressed only in adipose tissues, both white and brown. A ten-fold increase in ob mRNA was detected in white adipose tissues of obese animals compared to the lean ones of the JCR-LA:cp strain of rat. Ob gene was expressed in adipocytes and preadipocytes from the obese rat whereas in the lean and SD rats, ob gene expression was found in adipocytes only. No ob mRNA was detected in preadipocytes from the lean or SD rats, indicating a differentiation or maturation-dependent expression in normal rats. PMID- 7488083 TI - In vivo and in vitro binding of microcystin to protein phosphatases 1 and 2A. AB - The hepatotoxic microcystins (Mcyst) are potent inhibitors of the ser/thr protein phosphatases (PP1 and PP2A) with IC50's of 0.1-1.0 nM. Mcyst and other PP inhibitors like okadaic acid or calyculin A interact with the C-terminal region of PP1 and PP2A. Using [125I]-Mcyst and antibodies specific for PP1 and PP2A, we show by immunoprecipitation and autoradiography, that in hepatocytes Mcyst forms secondary covalent bonds with both PP1 and PP2A catalytic subunits. We demonstrate that the bond resulted from the reaction between the electrophilic alpha, beta unsaturated carbonyl of the methyldehydroalanine residue of Mcyst and the thiol of Cys 273 located in the C-terminal of PP1 (Cys 266 in PP2A), since site-directed mutagenesis of Cys 273 to Ala in PP1 alpha led to complete loss of ability for the formation of a covalent Mcyst-PP1 adduct. PMID- 7488084 TI - An alternately spliced mRNA encoding functional domains of murine MAdCAM-1. AB - cDNA clones representing a small (0.8 kb) form of murine mucosal addressin cell adhesion molecule-1 (MAdCAM-1) mRNA were obtained and sequenced. The sequence was identical to the published 1.6 kb murine MAdCAM-1 cDNA sequence, except that 432 nucleotides encoding the mucin-like and IgA-homologous portions were deleted. This cDNA most likely represents an alternately spliced mRNA. Substantial amounts of both the short and long MAdCAM-1 mRNAs are present in murine mesenteric lymph node. Ig fusion proteins displaying either the short or long forms of MAdCAM-1 can bind Mn(++)-activated JY cells bearing human alpha 4 beta 7 integrin, indicating that the two N-terminal Ig-like domains of MAdCAM-1 are sufficient to bind its integrin counter-receptor. PMID- 7488085 TI - Ulex europaeus I lectin induces activation of matrix-metalloproteinase-2 in endothelial cells. AB - In this report, we show that the lectin Ulex europaeus agglutinin I (UEA I), which binds to alpha-linked fucose residues on the surface of endothelial cells, mediates activation of the 72-kDa matrix metalloproteinase-2 (MMP-2). A dose dependent increase in the active 62-kDa form of MMP-2 was observed in conditioned medium from monkey aortic endothelial cells (MAEC) following incubation with concentrations of UEA I ranging from 2 to 100 micrograms/ml. The increase in the 62-kDa MMP-2 gelatinolytic activity was not reflected by a rise in MMP-2 gene expression. The UEA I-mediated activation of MMP-2 was blocked by L-fucose, which competes with UEA I for binding to alpha-fucose. These findings may suggest that a similar in vivo mechanism exists, whereby adhesive interactions between tumor cell lectins and endothelial cells can mediate MMP-2 activation. PMID- 7488086 TI - Effect of diabetic duration on the secondary structures of the human lens capsules in diabetic cataracts. AB - The decrease in triple helix content from 16% to 9% and in beta-turn content from 45% to 33%, combined with markedly increased random coil structure (16%) can be found in the secondary structure contents of lens capsule of the premature cataractous patients with longer diabetic history. However, almost the same peak position and secondary structure content are observed in the curve-fitted IR amide I band of lens capsules of both non-diabetic controls and premature cataractous patients with shorter diabetic history, even if the cataracts are senile. The present study emphasizes the duration of diabetes as a major cause to alter the secondary conformation of the cataractous human lens capsules in diabetic patients. PMID- 7488088 TI - Induction of liver cytochrome P450 2B1 by beta-ionone in Sprague Dawley rats. AB - Induction of liver cytochrome P450 2B1 by beta-ionone was investigated in male and female Sprague Dawley rats. Administration of beta-ionone subcutaneously 72 and 48 hr before sacrificing the animals not only significantly induced the liver microsomal activity of pentoxyresorufin O-dealkylase, but also clearly increased in the level of cytochrome P450 2B1 protein. The induction of cytochrome P450 2B1 by beta-ionone was much greater in male rats than in female rats. A slot blot analysis showed that the mRNA level was increased from 6 hr after treatment with beta-ionone in male rats and from 12 hr after treatment in female rats. Taken together, the present results indicate for the first time that the induction of cytochrome P450 2B1 by beta-ionone might be regulated by the accumulation of mRNAs. PMID- 7488087 TI - Adenylyl cyclase integrates multiple G protein signals to modulate calcium currents in neonatal rabbit heart. AB - We investigated the effects of added beta gamma subunits of G proteins (G beta gamma) on beta-adrenergic responsiveness of transmembrane Ca2+ currents (ICa) in ventricular myocytes from neonatal rabbits. G beta 1 gamma 1 purified from retinal rods was dialyzed into cells via the voltage clamp micro-electrode. Stimulation of ICa by isoproterenol was not affected by added intracellular G beta 1 gamma 1 or by carbachol alone but was completely blocked by combined G beta 1 gamma 1 and carbachol. Pretreatment of cells with pertussis toxin or temporal separation of carbachol and isoproterenol allowed stimulation of ICa by isoproterenol in cells dialyzed with G beta 1 gamma 1. Carbachol and G beta 1 gamma 1 together also did not prevent stimulation of ICa by dibutyryl-cyclic AMP. Thus, rather than simply inactivating Gs alpha by mass action, G beta 1 gamma 1 acts in concert with carbachol to inhibit isoproterenol stimulation of ICa. PMID- 7488089 TI - Galanin inhibits cholecystokinin secretion in STC-1 cells. AB - Neuropeptides such as gastrin releasing peptide and pituitary adenylate cyclase activating polypeptide (PACAP) stimulate CCK secretion from CCK producing cells. We hypothesized that in addition to somatostatin, galanin may also play an inhibitory role on CCK secretion. The effect of galanin on CCK secretion was studied in a CCK-producing murine neuroendocrine tumor cell line, STC-1. Galanin below 10 nM did not affect basal CCK secretion but dose- and time-dependently inhibited KCl-stimulated CCK secretion. Galanin also inhibited forskolin-, bombesin- and PACAP- but not dibutyryl cAMP- or beta-TPA-stimulated CCK secretion. The inhibitory effect of galanin was reduced partially by a blocker of ATP-sensitive K+ channel (K+ ATP), glibenclamide, and prevented by pretreatment of the cells with PTX. The results indicated galanin regulates CCK secretion by modulation of K+ ATP and cAMP production through receptors coupled to a PTX sensitive G protein. PMID- 7488091 TI - DAN gene product has an affinity for Ni2+. AB - Differential screening-selected gene aberrative in neuroblastoma (DAN) encodes a protein which possesses metal binding motifs. Glutathione S-transferase DAN fusion protein had an ability to bind to Ni(2+)-immobilized affinity resin. Truncation of the C-terminal region including a (HX)n repeat of DAN caused a loss of binding ability to the affinity resin, suggesting that this region is essential for Ni(2+)-binding. DAN produced in cultured rat cells also had an affinity for Ni2+. Cross-linking experiments demonstrated that the C-terminal region might function as a protein-protein interacting domain. PMID- 7488090 TI - Hyperphosphorylated p47-phox lost the ability to activate NADPH oxidase in guinea pig neutrophils. AB - p47-phox is one of the cytosolic activation factors of NADPH oxidase in neutrophils and known to translocate to plasma membranes and function by protein kinase C-phosphorylation. In cytosol fraction, prepared from calyculin A-treated neutrophils, the activity of cytosolic factor to activate NADPH oxidase was more reduced than that from PMA-treated cells. But, p47-phox did not translocate to the membranes, even if p47-phox was hyperphosphorylated in the calyculin A treated neutrophils. Such hyperphosphorylated p47-phox seemed to lose the activity to constitute NADPH oxidase complex. PMID- 7488092 TI - Upstream structure of human ADH7 gene and the organ distribution of its expression. AB - The ADH7 gene encoding human class IV (sigma) alcohol dehydrogenase (ADH) was cloned from a Caucasian genomic DNA library and its upstream structure was determined. Moreover, the organ distribution of its expression was examined. Northern hybridization analysis with a specific probe for sigma-ADH showed that expression of the gene is organ specific rather than ubiquitous, and occurs in the stomach but not in the liver. The lack of CG rich sequence and presence of TATA and CCAAT boxes in the upstream region of ADH7 may reflect the organ specific expression. The findings that this region lacks hepatocyte nuclear factor and has only one CCAAT enhancer binding protein consensus site may account for the fact that this gene is not expressed in the liver. The upstream region had sequences which are compatible with a glucocorticoid response element, a metal binding factor-1, and an active gene regulatory protein-1, suggesting that sex hormones, zinc, and retinoic acid may be involved in the regulation of the expression of this gene. PMID- 7488093 TI - Characterization of multiple metalloproteinases with fibrinogenolytic activity from the venom of Taiwan habu (Trimeresurus mucrosquamatus): protein microsequencing coupled with cDNA sequence analysis. AB - Three fibrinogenolytic proteases were isolated and purified from the venom of Taiwan habu (Trimeresurus mucrosquamatus) using anion-exchange and gel-filtration chromatographies followed by cation-exchange HPLC. Further characterization of these purified fractions with fibrinogenase activity indicated that they are single-chain proteases of approximately 24 kDa, possessing strong cleaving activity mainly on the A alpha and less on B beta and gamma chains of fibrinogen subunit chains. Enzyme activities were strongly inhibited by EDTA or 1,10 phenanthroline and not by phenylmethanesulfonyl fluoride, suggesting that these fibrinogenases belong to the family of metalloproteinases and not thrombin-like serine proteases. N-Terminal sequence analysis of these proteases failed to show any free amino-terminal residues, thus hampering the sequence determination by conventional sequencing strategy. Microsequencing on the electroblotted fragments of CNBr-treated proteases separated on SDS-PAGE was then used to determine the partial sequences. Sequence comparison of the determined partial sequences of these proteins with published sequences of the protein data bank revealed that they showed sequence homology with H2-protease. HR2a and protrigramin, which were all shown to belong to metalloproteinases present in various snake venoms. Polymerase chain reaction (PCR) was employed to amplify cDNAs constructed from the poly(A)+RNA of fresh venom glands of the same snake species to facilitate cloning and sequencing of these proteases. Sequencing several positive clones containing amplified cDNAs revealed the existence of one fibrinogenase in the Taiwan habu, which was contained within one complete cDNA encoding the preproproteinase precursor of hemorrhagic metalloproteinases. PMID- 7488094 TI - Maxadilan binds to membrane fractions of brain tissue. AB - Maxadilan is a potent vasodilator peptide isolated from salivary glands extracts of the hematophagous sand fly. Besides effects on the cutaneous vasculature, it has also been shown to relax rabbit aortic rings while elevating levels of cAMP. As a result of the effects on the skin and aorta, it was elected to undertake an examination of the tissue distribution of binding sites for maxadilan. In addition to specific binding in rabbit aorta and spleen, binding was detected in brain from various species including human, bovine, rabbit, rat and mouse with a kD of between 85 and 201 pM. Competitive displacement of [125I] maxadilan by a number of known vasoconstrictor peptides, vasodilator peptides and small molecule receptor ligands did not occur in the rabbit brain preparation. These results suggest the presence of specific binding sites in mammalian tissue for maxadilan whose endogenous ligand remains unknown. PMID- 7488095 TI - Identification of novel guanylyl cyclases from chemosensory tissues of rat and cattle. AB - A number of studies have shown that cGMP may play some roles in chemosensory transduction. To identify the structure of guanylyl cyclase in chemosensory tissues, cDNA fragments encoding guanylyl cyclase catalytic domain were amplified from rat and bovine olfactory and tongue epithelium using degenerate oligonucleotide primers and reverse transcription-polymerase chain reaction (RT PCR). Three novel clones, two membrane type guanylyl cyclases (RAT GC-1, BOV GC 3) and one soluble type guanylyl cyclase (RAT GC-2) were identified. RAT GC-1 was distributed over various rat tissues in addition to these chemosensory organs. BOV GC-3 was similar to but distinct from recent cloned olfactory-specific guanylyl cyclase. RAT GC-2 was identified as rat homologue of alpha 2 subunit of the soluble guanylyl cyclase. PMID- 7488096 TI - Regional expression and dietary regulation of rat small intestinal peptide and amino acid transporter mRNAs. AB - RT-PCR was used to obtain rat small intestinal cDNAs for two peptide transporters, showing conclusively for the first time that both are present in normal intestinal mucosa. Sequencing of these cDNAs showed them to be highly homologous and similar to two different types of peptide transport proteins from either colorectal carcinoma cells (Caco-2) or human and rabbit intestine. An even distribution profile of steady state levels of mRNA for both peptide transporters was observed along the longitudinal axis of small intestine. Both were upregulated in the distal regions of intestine by a high protein diet. Also, high levels of the rat high affinity glutamate transporter EAAC1 were observed in the distal intestine. These results suggest that the distal regions of small intestine play an important role in the absorption of some amino acids and peptides. Furthermore this area appears to be a primary site where dietary induced changes in peptide and amino acid transport occurs. PMID- 7488097 TI - Gamma-glutamylcysteine synthetase gene overexpression results in increased activity of the ATP-dependent glutathione S-conjugate export pump and cisplatin resistance. AB - The ATP-dependent glutathione S-conjugate export pump (GS-X pump) has been suggested to play a role in the mechanism of cisplatin resistance. The purpose of this study was to determine the relationship between intracellular glutathione (GSH) levels and GS-X pump activity and whether GS-X pump overexpression results in cisplatin resistance. We transfected the human gamma-glutamylcysteine synthetase (gamma-GCS) gene into a human small-cell lung cancer cell line, SBC-3, producing SBC-3/GCS. The intracellular GSH content of SBC-3/GCS was twice that of the parental line, its GS-X pump activity was significantly enhanced and cellular cisplatin accumulation decreased. SBC-3/GCS showed higher resistance (relative resistance value of 7.4) to cisplatin than the parental line SBC-3. These data indicate that gamma-GCS gene overexpression induces cellular cisplatin resistance associated with increases in both the GSH content and GS-X pump activity, resulting in reduced cisplatin accumulation. In conclusion, GS-X pump expression is related to cellular GSH metabolism and involved in cisplatin resistance. PMID- 7488098 TI - Truncated type II receptor for BMP-4 induces secondary axial structures in Xenopus embryos. AB - BRK-3 is a vertebrate type II receptor for BMP-4 distantly related to invertebrate type II receptors for BMP-2/BMP-4/dpp, such as daf-4 and punt. BRK-3 has a long carboxy-terminal sequence following intracellular kinase domain and is capable of forming a high-affinity complex with a type I receptor, BRK-2. To examine the role of BRK-2 + BRK-3 receptor complex in BMP signaling during early embryogenesis, the dominant-negative form of BRK-3 was ectopically expressed in the Xenopus embryos. A secondary body axis expressing the Sonic hedgehog and N CAM genes is induced by injecting mRNA encoding truncated form of BRK-3 into ventral marginal region, implicating the BMP signaling in axial mesoderm induction. Formation of the secondary axis depends on whether the deletion extends into the kinase domain, not into the carboxy-terminal tail, suggesting that the kinase domain, but not the tail region, is essential for BMP signaling. PMID- 7488099 TI - Cloning and expression of human uridine phosphorylase. AB - Using a mouse cDNA probe we have identified a human uridine phosphorylase cDNA clone from the cDNA library of a human colorectal tumor cell line, HCT116. The recombinant human uridine phosphorylase expressed in COS-7 cells demonstrated specific enzyme activity with uridine as the substrate; this activity was inhibited by the competitive inhibitor 2,2'-anhydro-5-ethyluridine. Northern blot analysis with the cDNA as a probe demonstrated high levels of mRNA expression in several tumor cell lines but very low level in normal cell, WI-38. The expression of uridine phosphorylase mRNA in HCT-116 cells was further enhanced by treating the cells with vitamin D3 and the inflammatory cytokines: tumor necrosis factor alpha, interleukin 1 alpha and interferon gamma. PMID- 7488100 TI - RNA polymerase II transcribes Dictyostelium untranslatable gene, dutA, specifically in the developmental phase. AB - In Dictyostelium discoideum, a novel type of RNA (dutA RNA), which is untranslatable, cytosolic and 1.3 kb in size, appears specifically after the aggregation stage [Yoshida, H. et al. (1994) Nucleic Acids Res. 22, 41-46]. We here show that the dutA gene is transcribed by RNA polymerase II, based on its alpha-amanitin sensitive nature of in vitro transcriptional activity. We also show that the stage-specific accumulation of dutA RNA is primarily due to the stage-specific enhancement of the transcriptional activity of the gene. PMID- 7488101 TI - The cytosolic chaperonin subunit TRiC-P5 begins to be expressed at the two-cell stage in mouse embryos. AB - The cytosolic chaperonin TRiC is a large protein complex involved in the folding of newly synthesized actin and tubulin. The fertilization of the mouse oocyte is followed by a remodelling of the actin and tubulin filaments. The TRiC subunit TCP1 is expressed only from the 4-cell stage on, even though actin and tubulin are synthesized in the previous stages. We investigated the onset of synthesis of another subunit, TRiC-P5, during early mouse embryogenesis. We report that TRiC P5 is synthesized at the 2-cell stage in an alpha-amanitin sensitive manner. Thus, it is expressed before TCP1 and is one of the first proteins to be synthesized after zygotic genome activation. PMID- 7488102 TI - Ornithine decarboxylase induction during liver regeneration in IRS-1-deficient mice. AB - We investigated the induction of ornithine decarboxylase during liver regeneration after partial hepatectomy in IRS-1-deficient mice. There were no significant differences in ODC activity or the time course of changes in ODC activity between IRS-1-deficient mice and wild-type mice. PI 3'-kinase activity showed similar increases in both groups of mice. Furthermore, ODC induction in IRS-1 transfected CHO cells was studied after stimulation by addition of FCS. The maximal ODC activity was 2.5-fold greater in IRS-1-transfected CHO cells than in control CHO cells. Our results suggest that the IRS-1 pathway may be involved in ODC induction. The absence of a difference in ODC and PI 3'-kinase activity in the regenerating liver between IRS-1-deficient mice and wild-type mice may have been related to the compensatory effects of IRS-2/pp190 [Araki et al. Nature (1994) 372, 186-190; Tobe et al. J.Biol.Chem. (1995) 270, 5698-5701]. PMID- 7488103 TI - Neuronal differentiation of Neuro 2a cells by lactacystin and its partial inhibition by the protein phosphatase inhibitors calyculin A and okadaic acid. AB - Lactacystin (1.3 microM), a metabolite from an actinomycete, induced the formation of bipolar projections at both sides of the cell body of Neuro 2a cells 1 day after treatment and networks at and after 3 days and enhanced acetylcholinesterase activity (a marker of neuronal differentiation). Thus, the neuronal differentiation was characterized both morphologically and functionally. The experiments with various inhibitors of protein kinases and phosphatases revealed that the protein phosphatase inhibitors calyculin A (0.5 nM) and okadaic acid (0.6 nM) inhibit the formation of bipolar projections at 1 day, but does not inhibit the network formation at and after 3 days. PMID- 7488104 TI - Hepatocyte growth factor stimulates wound repair of the rabbit esophageal epithelial cells in primary culture. AB - We have recently established an in vitro primary culture system for esophageal epithelial cells, which enabled us to investigate the effect of hepatocyte growth factor (HGF) and other factors on esophageal restitution. HGF remarkably stimulated restitution of these cells. So did epidermal growth factor (EGF), though moderately. Restitution velocity of esophageal cells was remarkably higher than that of gastric epithelial cells. The expression of c-met, specific HGF receptor was demonstrated by the esophageal cells, suggesting that the effect of HGF was mediated by its specific receptor. The expression level of c-met mRNA was the same as that of gastric epithelial cells, as assessed by competitive RT-PCR technique. These results suggest that HGF might be involved in the repair process of esophageal mucosal damage. PMID- 7488105 TI - Domain III exchanges of Bacillus thuringiensis CryIA toxins affect binding to different gypsy moth midgut receptors. AB - Aminopeptidase-N, purified from gypsy moth (Lymantria dispar L.) brush border membrane vesicles, exhibited specific binding to CryIAc toxin but not to CryIAa toxin. CryIAa-CryIAc hybrid toxins were used to localize the aminopeptidase-N binding region on CryIAc. Slot blot assays and ligand blot experiments demonstrated that the hybrid toxins which have the residues 451 to 623, comprising essentially domain III, from CryIAc toxin exhibited strong binding to purified aminopeptidase-N and 120 kDa brush border membrane protein. In contrast, the hybrid toxins which have the residues 451 to 623 from CryIAa toxin failed to bind to aminopeptidase-N, but did bind to another receptor, a 210 kDa protein. This is the first direct evidence that domain III is involved in receptor binding and the first to demonstrate that domain III substitutions direct the binding of these toxins to different gypsy moth midgut receptors. PMID- 7488106 TI - Non-oxidative loss of glutathione in apoptosis via GSH extrusion. AB - Reduced glutathione (GSH) has been hypothesized to play a role in the rescue of cells from apoptosis, by buffering an endogenously induced oxidative stress. We correlated GSH levels and apoptosis in U937 human monocytic cells induced to apoptosis by different agents. All treatments led to depletion of GSH concomitant with the onset of apoptosis. The loss was due to extrusion of GSH outside the cell, while GSSG was not accumulated in the apoptosing cells, nor was it found in the extracellular medium. Modulation of intracellular GSH level did not influence the overall extent of apoptosis. We conclude that glutathione loss in apoptosis is not necessarily preceded by an oxidative stress, and that GSH depletion alone is not sufficient to lead cells to apoptosis. PMID- 7488107 TI - Cloning of a cDNA encoding a 190-kDa insulin receptor substrate-1-like protein of simian COS cells. AB - Major insulin signals such as stimulation of glucose uptake and DNA synthesis and modification of hexose metabolism are mediated by the tyrosine-phosphorylated insulin receptor substrate-1 (IRS-1; pp180) in many species of cells. We cloned cDNA encoding a 190-kDa IRS-1-like protein (pp190) in simian COS cells and which is slightly larger than IRS-1 (pp180) of human, rat, and mouse cells. The deduced amino acid sequence of COS pp190 consisted of 1251 amino acids and was 96.4%, 87.9% and 88.7% identical to human, mouse and rat IRS-1. The COS pp190 bound to SH2 (src-homology 2) domains of p85, Grb2/Ash, and SH-PTP2, as did IRS-1. In IRS 1-knockout mice, insulin signals are thought to be mediated by IRS-2 (pp190), which is an alternative signaling molecule and is slightly larger than IRS-1. However, the COS pp190 may be a simian homologue of IRS-1, but not of IRS-2. The results of Southern blotting suggested the possibility that Chinese hamster ovary (CHO) cells have not only the IRS-1 gene but also a gene related to the COS pp190. PMID- 7488108 TI - Stabilisation of purified human collagenase by site-directed mutagenesis. AB - During purification, human fibroblast collagenase breaks down into two major forms, an N-terminal 22000/25000-Mr fragment and a C-terminal 27000-Mr fragment; the most likely mechanism being autolysis. The cleavage site has been identified (Pro269- Ile270) and in an attempt to obtain full-length human collagenase (i.e., Mr 42570), this cleavage site and another potential cleavage site (Ala258- Ile259) have been mutated by PCR- directed mutagenesis: Ile270Ser and Ile259Leu. The mutated cDNA was then cloned into the expression vector, pGEX2T, and expressed in Escherichia coli as a fusion protein with glutathione-S-transferase (GST). After cleavage with factor Xa, the mutated collagenase was purified on a peptide hydroxamic acid affinity column. The mutated recombinant collagenase is stable, remains full length and retains the ability to cleave collagen. PMID- 7488109 TI - TPO/c-mpl ligand induces tyrosine phosphorylation of multiple cellular proteins including proto-oncogene products, Vav and c-Cbl, and Ras signaling molecules. AB - Thrombopoietin (TPO)/c-mpl ligand is a hematopoietic growth factor that stimulates proliferation and maturation of megakaryocytes. To analyze the signaling pathway downstream of the c-mpl product, we used a human megakaryoblastic cell line, Mo7e, that has been proved to be responsive to TPO in terms of DNA synthesis. In this study, we found that TPO treatment resulted in tyrosine phosphorylation of Jak-2 kinase. Moreover, it was revealed that several functional molecules involved in the Ras signaling pathway, Shc and Sos, were phosphorylated by treatment with TPO. Finally, tyrosine phosphorylation of the proto-oncogene products, Vav and c-Cbl, has been proved to be induced by TPO. These results suggest that TPO could activate several signaling pathways including the Jak/Stat pathway, the Ras pathway and possibly another pathway involving the c-Cbl proto-oncogene product. PMID- 7488110 TI - Molecular cloning and sequencing of the murine gastrin gene. AB - Gastrin is a peptide hormone important in acid regulation, growth of enterochromaffin cells of the oxyntic mucosa, and smooth muscle contractility. We isolated a genomic clone of gastrin from a SV-129 murine genomic library. The murine gastrin gene contains an open reading frame of 101 amino acids. The deduced amino acid sequence has 91.1% homology with the rat and 67.3% homology with the human analogue. Southern blot analysis of the murine gastrin clone gave the same restriction pattern as that seen from mouse kidney genomic DNA, consistent with this being a single copy of the gastrin gene in the mouse genome. Transfection studies demonstrated that the murine gastrin gene expression is stimulated by epidermal growth factor to the same extent as the human gastrin gene. PMID- 7488111 TI - The molecular basis of the self/nonself selectivity of a coelenterate toxin. AB - Coelenterates produce potent hemolysins inhibited by sphingomyelin (SM). Remarkably, instead of this lipid, their membranes contain a phosphono analogue of it. Using coelenterolysin (CL), a toxin produced by the sea anemone Phymactis clematis, we have examined a possible connection between these two peculiar traits. Our experiments showed that, while SM binds this lysin and inhibits its hemolytic activity, the endogenous PnSL do neither. In addition, liposomes made of bovine erythrocyte lipids are rapidly disrupted by CL, while those made of P. clematis lipids are completely resistant to it. However, if small amounts of SM are added to the P. clematis lipids, the resulting liposomes become sensitive to CL. Taken together, our results show for the first time that substitution of SM by its phosphono analogue is the molecular basis for the selectivity of an anthozoan toxin. We therefore propose that exotoxin production and membrane composition are coadapted traits that confer on the coelenterates a significant evolutionary advantage. PMID- 7488112 TI - Augmented expression of the obese gene in the adipose tissue from rats fed high fat diet. AB - Expression of the obese (ob) gene is augmented in the adipose tissue in several rodent models of genetic obesity. In the present study, we examined the ob gene expression in a rodent model of acquired obesity obtained by pure overfeeding of normal rats. Male Sprague-Dawley rats at 8 weeks of age were fed standard diet or high-fat diet. Rats fed high-fat diet developed moderate degree of obesity, hyperglycemia, and hyperlipidemia as compared with rats fed standard diet. Northern blot analysis revealed that the ob gene is expressed abundantly in the adipose tissue obtained from the epididymal, mesenteric, subcutaneous, retroperitoneal, and interscapular fat pads in rats fed standard diet. Expression of the ob gene was augmented in all the adipose tissue examined in rats fed high fat diet. The present study demonstrates that the ob gene expression is augmented in the adipose tissue in diet-induced obesity, thereby suggesting the pathophysiologic roles of the ob gene in acquired obesity. PMID- 7488113 TI - Gene transcription of angiotensin II type 2 receptor is repressed by growth factors and glucocorticoids in PC12 cells. AB - The rat angiotensin II type 2 receptor (AT2-R) expression was markedly downregulated by the mitogenic action of serum, growth factors and dexamethasone. The regulation by serum or growth factors did not affect the AT2-R mRNA half-life (18 h), whereas the AT2-R half-lives of dexamethasone-treated cells and proliferating cells decreased to 10 h and 15 h, respectively. Nuclear run-off assays indicated the mechanism of repression of AT2-R expression by serum, growth factors and dexamethasone or in proliferating cells to be, in large part, transcriptional. These findings indicate that transcription of the AT2-R gene is regulated in a growth state-dependent manner and suggest that this regulation provides a means by which cells can modulate their responsiveness to the actions of angiotensin II mediated through AT2-R. PMID- 7488114 TI - Endotoxin antagonism by a synthetic lipid A analogue, DT-5461, with low endotoxicity in human peripheral blood monocytes. AB - We examined the molecular mechanism of DT-5461-induced LPS antagonism in human peripheral blood monocytes. Dose-response studies revealed that LPS-induced IL-1 and TNF-alpha production was apparently totally suppressed in a competitive manner by a 10-fold excess of DT-5461. A 10-fold excess of DT-5461 significantly blocked the binding of FITC-LPS to the monocytes. DT-5461 suppressed IL-1 and TNF alpha mRNA expression in LPS-activated monocytes. Western blots showed that DT 5461 suppressed the LPS-induced tyrosine phosphorylation of p42mapk/ERK2. These results suggested that the competitive binding inhibition and repression of early intracellular signaling involved in DT-5461-mediated LPS antagonism. PMID- 7488115 TI - A designed beta-hairpin peptide. AB - A synthetic octapeptide, Boc-Leu-Val-Val-D-Pro-Gly-Leu-Val-Val-OMe (1) has been designed as a model for a beta-hairpin conformation. Circular dichroism spectra in various organic solvents reveal a single negative band at 214-217 nm consistent with beta-sheet structures. NMR studies in CDCl3 and C6D6 establish the solvent shielded nature of the Leu(1), Val(3), Leu(6) and Val (8) NH groups. Nuclear Overhauser effects are observed between Val(7) C alpha H and Val(2) C alpha H protons providing strong support for a beta-hairpin conformation. Several important diagnostic interresidue NOEs establish a Type II' beta-turn conformation for the D-Pro-Gly segment and extended conformations for the amino and carboxyl terminal tripeptide arms. The high solubility of the beta-hairpin peptide in organic solvents holds promise for the development of models for three and four stranded beta-sheets. PMID- 7488116 TI - Cloning and characterization of seizure-related gene, SEZ-6. AB - SEZ-6 is a brain-specific cDNA. Its expression is increased by convulsant drug and it encodes membrane protein with five copies of short consensus repeat (SCR; complement C3b/C4b binding site) and two repeated sequences which are partially similar to CUB domain (complement Clr/s-like repeat). In this study, we analyzed the gene product of SEZ-6 by antibody raised against SEZ-6 peptide. In vitro translation and immunoblot revealed that SEZ-6 protein is modified post translationally and expressed in the cerebrum and the cerebellum. To analyze the SEZ-6 expression in detail, we searched for other types of SEZ-6 cDNA by the polymerase chain reaction. Two additional types of cDNAs were isolated and sequence analysis showed that one cDNA encodes membrane protein with a different C-terminal region and the other encodes secreted protein with two SCRs and one CUB-like domain. PMID- 7488117 TI - Cloning of the cDNA for a brain glycine-, glutamate- and thienylcyclohexylpiperidine-binding protein. AB - Polyclonal antibodies (Ab's) were raised against a 43-kDa component of a protein complex that has ligand recognition sites similar to those of brain N-methyl-D aspartate (NMDA) receptors. The Ab's were used to immunopurify from brain synaptic membranes a 60-kDa glycine (Gly), glutamate (Glu) and thienylcyclohexylpiperidine (TCP)-binding protein and to screen a rat hippocampal cDNA expression library. A 1.85-kb clone, pGlyBP, coding for a protein of 470 amino acids (52.7 kDa) was identified. Northern blot analyses performed on poly(A+) RNA from brain revealed hybridization of the labeled cDNA probes to transcripts of 1.9 kb. E. coli transformed with the pGyBP expressed a protein that was recognized by the anti-43 kDa Ab's and had recognition sites for Gly, Glu and TCP. The cloned protein has 2 glycosylation sites, 3 hydrophobic domains, 4 cysteine-rich motifs (C-X2-C-X16-20-C-X5-11), and 2 regions homologous to the NR1 receptor protein. PMID- 7488120 TI - Noncovalent interactions in maintaining the native structure of low density lipoprotein. AB - The integrity and the surface charge distribution of native low density lipoprotein (LDL) are prerequisites of its binding to the LDL receptor. Oxidation is one of the most important physiological effects resulting in an altered structure and metabolism of LDL. To reveal forces responsible for maintaining the intact structure of LDL in the absence of cells we have determined the kinetics of lipid peroxidation, changes in electrophoretic mobilities and size distributions of LDL samples as a function of Cu++ induced oxidative modification in a cell-free system at two different (50 mM and 150 mM) ionic strengths by electrophoretic and dynamic light scattering. Our data show that the lipid peroxidation is almost complete before LDL is degraded at 50 mM while a slight extent of lipid peroxidation is enough to result in the same effect at 150 mM. These suggest that both ionic and hydrophobic interactions are necessary to maintain the integrity of the LDL molecule. PMID- 7488119 TI - Recombinant human pancreatic ribonuclease produced in E. coli: importance of the amino-terminal sequence. AB - Human pancreatic ribonuclease 1 (hRNase 1) in the mature form has been produced in E. coli using T7 expression system. The recombinant hRNase 1 protein was solubilized from the inclusion bodies, refolded in glutathione redox system, and purified through chromatographic procedures by utilizing cation-exchange and reversed-phase columns. The ribonucleolytic activity of recombinant hRNase 1 was examined on yeast RNA and cytidylyl-3',5'-adenosine revealing the distinctive ribonucleolytic activity. The activity was perfectly inhibited by human placental RNase inhibitor. Truncation of 7 amino acid residues in the amino-terminal sequence resulted in much reduction in ribonucleolytic activity and in affinity to human placental RNase inhibitor with the disintegration of secondary structures of the protein observed by circular dichroism spectra. The present study has revealed the important contribution of the amino-terminal sequence of hRNase 1 to the characteristics of the protein. PMID- 7488118 TI - Receptor-linked antigen delivery system. Importance of autologous alpha 2 macroglobulin in the development of peptide vaccine. AB - We have hijacked a process of the receptor-mediated endocytosis to transport peptide antigens into antigen presenting cells (APCs) for the purpose of increasing the level of antigen presentation (named Receptor-Linked Antigen Delivery System (R-LADS)). By coupling an endogenous plasma proteinase inhibitor alpha 2-macroglobulin (alpha 2M) to a synthetic peptide having a partial sequence of HIV-1 envelope protein, alpha 2M was made to carry the peptide into APCs as a part of the normal alpha 2M cycle, which resulted in an increased production of specific antibodies against the peptide (Mitsuda, S., Nakagawa, T., Osada, T., Shimamoto, T., Nakazato, H. and Ikai, A. (1993) Biochem. Biophys. Res. Commun. 194, 1155-1160). We demonstrate here that this procedure becomes a more efficient tool for antibody production when autologous transporter protein was used. By using murine alpha 2M (m alpha 2M) instead of heterologous human alpha 2M (h alpha 2M) when mice were experimental animals, we were able to dramatically enhance the production level of anti-HIV-1 peptide antibodies and shorten the period which is needed for antibody production. We aim to develop effective peptide vaccines by further improving this system. PMID- 7488121 TI - Nucleoside triphosphate donors for nucleoside kinases: donor properties of UTP with human deoxycytidine kinase. AB - The reported higher efficiency of UTP, relative to ATP, as phosphate donor for deoxycytidine kinase (dCK), has been extended and found to apply to both dCyd and dAdo as acceptors. UTP as phosphate donor was shown to follow strictly Michaelis kinetics, with Km = 1 microM, in striking contrast to ATP, which exhibits marked negative cooperativity (Hill coef. = 0.7) with a several-fold higher Kmapp = 15 microM. Phosphate transfer was followed directly with use of mixtures of [gamma 32P]ATP and cold UTP as donors, or with 3H-labeled acceptors and cold donors. With equimolar concentrations of ATP and UTP (50 microM or 1 mM each), and dCyd or dAdo as acceptor, only minimal phosphate transfer occurred from ATP (3-10%). With a 6:1 ratio of ATP:UTP, hence exceeding the intracellular ratio, phosphate transfer from ATP increased, but still did not exceed 25-40% with either dCyd or dAdo as acceptor. Moreover, relative ATP transfer is dependent on the dCyd concentration. We conclude that the major intracellular phosphate donor for dCK is not ATP, but UTP. Preliminary data for human thymidine kinases (TK1 and TK2) exhibit quite different behaviour. The foregoing, together with literature data, are highly relevant to in vitro studies on the properties of the nucleoside kinases, and to the design of chemotherapeutically active nucleoside analogues. PMID- 7488122 TI - Suppression of cyclin D1 but not cdk4 or cyclin A with induction of melanoma terminal differentiation. AB - To identify cyclins specifically associated with control of melanoma cell proliferation, we now compared expression of cyclin A, reported to be a marker for hematological malignancies, with that of cyclin D and its cdk4 kinase partner. All these proteins were expressed in proliferating B16 melanoma. However, L-tyrosine which induces melanoma terminal differentiation, selectively decreased cyclin D with no comparable effect on cdk4 or cyclin A. A 2-hour exposure of the cells to the tyrosine phosphatase inhibitor, sodium vanadate, further decreased cyclin D from differentiated cells, suggesting that tyrosine phosphorylation regulates cyclin D turnover. Addition of serum to starved cells also revealed that tyrosine did not block the early cyclin D increase associated with serum stimulation, but accelerated its subsequent loss. Our data suggest that cyclin D decrease with melanoma terminal differentiation could be an alternative mode of growth arrest even in cells harbouring a mutant or transcriptionally silent cdk4 inhibitor tumor suppressor p16ink4 gene. These results also imply that cyclin D may be useful as a target and as a prognostic marker in melanoma therapy. PMID- 7488123 TI - Human satellite cell proliferation in vitro is regulated by autocrine secretion of IL-6 stimulated by a soluble factor(s) released by activated monocytes. AB - We previously showed that macrophages, besides their scavenger role, selectively induce rat myoblast proliferation in vitro by releasing soluble factors. In this paper we demonstrate a relationship between human-activated monocytes and increased human myoblast proliferation due to IL-6 autocrine secretion by satellite cells. Indeed in the supernatants of muscle cultures treated with activated monocyte-conditioned medium we show by means of an ELISA quantitation a higher autocrine secretion of IL-6 associated with increased myoblast proliferation. This suggests that a growth factor(s) secreted by activated monocytes stimulates IL-6 production by myoblasts and then regulates proliferation of satellite cells. PMID- 7488124 TI - The liver response to in vivo heat shock involves the activation of MAP kinases and RAF and the tyrosine phosphorylation of Shc proteins. AB - We have investigated the mechanisms of signal transduction in the response of liver to heat shock in vivo. By immunoblot experiments we have shown that heat shock decreases the electrophoretic mobility of the 40 and 43 kDa mitogen activated protein kinases (MAPKs) and we have found a significant increase of MAPK activity measured as phosphotransferase capacity of both cytosolic extracts and MAPK immunoprecipitates. To elucidate the signalling pathway which accounts for MAPK activation, we focused our attention on its upstream factors, Raf and Ras. We have shown that, heat shock activates Raf-1 kinase and causes an increase in phosphotyrosine content of the 52 kDa Shc protein accompanied by an increment in the amount of coimmunoprecipitated Grb2. These findings provide the first evidence that the Ras-Raf-MAPK pathway is activated in liver during heat shock in vivo. PMID- 7488125 TI - Ligand binding kinetics of a soluble full-length murine erythropoietin receptor. AB - The full-length murine erythropoietin receptor was expressed in Sf9 cells using a baculovirus vector. Erythropoietin receptors in solubilized Sf9 cell lysates bound erythropoietin with high affinity (92 pM). Erythropoietin receptor-125I labeled erythropoietin association and dissociation kinetics using solubilized Sf9 cell lysates revealed a ka of 0.16 nM-1 min-1 and a kd of 0.00055 min-1 giving an observed KD of 3.45 pM. The erythropoietin receptors was partially purified from Sf9 cell lysates by chromatography on Con A Sepharose. When erythropoietin receptors were crosslinked to 125I-labeled erythropoietin and analyzed by SDS-7.5% PAGE protein complexes of 90 and 125 kDa were observed with receptors in solubilized lysate, and 170 and 190 kDa with the partially purified receptors. PMID- 7488126 TI - Characterization of a silencer that modulates transcription of the human distal aldolase A promoter. AB - Transcription from the distal promoter (pL) of the human aldolase A gene is driven by both positive and negative cis-acting elements. With footprinting and gel mobility assays we defined: (i) the position of the negative regulatory cis element (AldA-NRE), which spans a GA-rich sequence; (ii) the sequence of AldA NRE, which is critical for protein specific binding and is homologous to other silencer-like motifs. Removal of the negative cis-element fully restores the expression of the distal promoter. Insertion of the AldA-NRE upstream from a heterologous promoter results in a 6-fold decrease in CAT reporter gene expression, suggesting that AldA-NRE might be involved in a more general type of mechanism that mediates repression of gene transcription. PMID- 7488127 TI - Cloning and characterization of the murine activin receptor like kinase-1 (ALK-1) homolog. AB - Multiple serine/threonine kinases were detected in a bone marrow stromal cell line. One of these, the murine activin receptor like kinase-1 (ALK-1) homolog was cloned and sequenced. The expressed recombinant protein (62 kDa) was consistent in size with that predicted by the cDNA (58.6 kDa). On Western blots, a goat polyclonal antibody detected the native ALK-1 protein in bone marrow stromal cells, lung, brain, kidney and spleen. Two protein species of 60 kDa and 72-76 kDa were detected. Glycosylation events or alternative splicing may account for the larger protein specie. PMID- 7488128 TI - Decrease in actin gene expression in melanoma cells compared to melanocytes is partly counteracted by BrdU-induced cell adhesion and antagonized by L-tyrosine induction of terminal differentiation. AB - Malignant transformation is frequently accompanied by changes in the cytoarchitecture of adherent cells, which may be influenced by fluctuations in actin gene expression. We now show that normal melanocytes express a 5 fold higher level of actin mRNA than their melanoma counterparts. Induction of terminal melanogenesis did not increase actin in melanoma cells. However, culture with the thymidine analog, Bromodeoxyuridine, increased actin expression in undifferentiated but not in differentiating melanoma. Cell detachment assays and cell shape comparisons revealed a direct correlation of actin mRNA with increased melanoma cell adhesion rather than with differentiation-mediated suppression of tumor growth. PMID- 7488129 TI - Phosphorylation of myosin-I from rat liver by protein kinase C reduces calmodulin binding. AB - Three isoforms of the cytoskeletal-associated, mechanochemical enzymes known as myosin-I have been purified from rat liver; each coisolates with calmodulin. Incubation of the purified myosin-I's with protein kinase C gamma and 32P-ATP results in phosphorylation of the myosin-I heavy chains. After phosphorylation, the myosin-I isoforms bind less radiolabeled calmodulin in binding assays than observed for control samples. Since the purified isoforms are phosphoproteins as determined by immunoblotting with monoclonal antibodies which recognize phosphoamino acids, these results indicate that phosphorylation might play a role in regulation of myosin-I. PMID- 7488130 TI - IL-1 induces expression of WAF1 mRNA in human fibroblasts: mechanisms of accumulation. AB - WAF1 inhibits cyclin-cyclin dependent kinase complexes, causing arrest in the cell cycle. We report that interleukin-1 (IL-1) regulates the levels of WAF1 protein and mRNA in human fibroblasts. Further studies showed that 12-O tetradecanoyl phorbol 13-acetate (TPA) enhanced the accumulation of WAF1; cells refractory to TPA still increased their levels of WAF1 mRNA when exposed to IL-1. IL-1 also increased transcription of WAF1 by 13-fold and WAF1 mRNA became 2-fold more stable in cells stimulated with IL-1 (t1/2 = 2 h) as compared to untreated cells (t1/2 = 1 h). In summary, IL-1 induces expression of WAF1, and this occurs by increased transcription of WAF1 and stabilization of WAF1 mRNA. Our data also indicate that the induction of WAF1 mRNA by IL-1 occurs through a pathway independent of protein kinase C activation. PMID- 7488131 TI - Interleukin 1 beta-induced expression of nitric oxide synthase in rat renal mesangial cells is suppressed by cyclosporin A. AB - The expression of inducible nitric oxide synthase (iNOS) is triggered in rat renal mesangial cells by exposure to the inflammatory cytokine interleukin 1 beta (IL-1 beta). Here we report that cyclosporin A (CsA) a potent immunosuppressive drug, inhibits IL-1 beta dependent iNOS expression in renal mesangial cells. Addition of CsA dose dependently suppresses IL-1 beta-induced nitrite formation (IC50 = 0.9 microM). Western- and Northern blot analyses of mesangial cell extracts reveal that the inhibition of IL-1 beta-induced nitrite formation by CsA is due to decreased iNOS protein and iNOS mRNA steady state levels. Using nuclear run on experiments we show that the transcription rate of the IL-1 beta-induced iNOS gene is reduced. Furthermore, by electrophoretic mobility shift analysis we demonstrate reduced DNA-binding of the nuclear factor NF kappa B, an essential component of the IL-1 beta-dependent upregulation of iNOS gene transcription. The data presented in this report suggest that the cellular machinery involved in the IL-1 beta dependent transcriptional upregulation of the iNOS gene in mesangial cells is a target for the action of CsA. PMID- 7488132 TI - Association of X-linked adrenoleukodystrophy with HLA DRB1 alleles. AB - Inflammatory demyelination in the central nervous system in the childhood cerebral phenotype of X-linked adrenoleukodystrophy (X-ALD) bears resemblance to that of multiple sclerosis. With a view to an association of HLA class II genes, specifically HLA-DRB1 subtype DRB1*15 to multiple sclerosis we investigated the HLA class II DR haplotype in 29 unrelated X-ALD patients including 17 childhood cerebral phenotype patients. Our results did not show an association of DRB1*15 and X-ALD, but disclosed a significant association of HLA DRB1*16 alleles and X ALD in general. This finding suggests that in addition to the X-chromosomal ALD gene an autosomal gene linked to the HLA class II region is involved in the pathogenesis of X-ALD. This gene should affect a pathomechanism common to all ALD variants, such as defective peroxisomal metabolism of very long chain fatty acids. PMID- 7488134 TI - Identification of a low specificity, oxygen, heme, and growth phase regulated DNA binding activity in Saccharomyces cerevisiae. AB - Cytochrome c oxidase from Saccharomyces cerevisiae is subject to intricate control at the level of transcription of the various genes encoding its subunits. Expression of the subunit VI encoding gene, COX6, is glucose repressed, growth phase induced, and dependent on oxygen and heme availability. An upstream activation region for COX6, UAS6, was found to contain a glucose responsive region, a heme dependent region (HDS1), and a binding site for the transcription factor, BAF1. BAF1 was the only factor observed to form a protein complex with UAS6 in vitro. However, we found that binding of BAF1 was unaffected by oxygen or heme regulation. In the present communication, we have identified a DNA binding activity that was growth phase induced and dependent on oxygen and heme availability. This highly regulated activity was detected by its discrete binding to the heme responsive site, HDS1, in UAS6. Nonetheless, it appears to have weak DNA binding specificity. PMID- 7488135 TI - Monoclonal antibody against the polymorphic site distinguishes apolipoprotein E4 from other isoforms. AB - Apolipoprotein E4 has been confirmed as a genetic risk factor for Alzheimer's disease. Although several hypotheses have been advanced to explain how the inheritance of apolipoprotein E isoforms affects the rate of Alzheimer's disease expression, the mechanism whereby apolipoprotein E is involved in the pathogenesis of Alzheimer's disease is still uncertain. To clarify the way in which the apolipoprotein E4 isoform differs from the others, we generated a monoclonal antibody specifically reactive with the apolipoprotein E4 isoform. This antibody suggests that the polymorphic site is important in differentiating the ApoE4 isoform from others. PMID- 7488133 TI - Mutations of the transforming growth factor-beta type II receptor gene and genomic instability in hereditary nonpolyposis colorectal cancer. AB - To determine the relation between the mutation of the TGF-beta type II receptor gene and genomic instability in the tumorigenesis of hereditary nonpolyposis colorectal cancer (HNPCC), we screened genomic DNA of 38 tumors from 25 HNPCC patients, 15 colorectal cancers from familial adenomatous polyposis patients, and 8 sporadic endometrial cancers, in two areas containing a (A)10 repeat or a (GT)3 repeat of the gene. Seventeen of the 24 (71%) genomic instability-positive HNPCC tumors carried one or two A deletions in the (A)10 repeat, while none of the 14 genomic instability-negative tumors did. These deletions inactivate the receptor through a frameshift mutation and the resultant protein truncation. No mutation was detected in the (GT)3 repeat sequence, but we found a missense mutation of codon 537 in the same area in one tumor. One A deletion was also detected in a genomic instability-positive sporadic endometrial cancer, but none in familial adenomatous polyposis tumors. No mutations were detected in the corresponding normal cells of these cases, indicating a somatic mutation. These data suggest that the TGF-beta type II receptor gene is a major target of genomic instability in HNPCC tumorigenesis. PMID- 7488136 TI - 17-beta estradiol protects neurons from oxidative stress-induced cell death in vitro. AB - The potential antioxidant activity of 17-beta estradiol and other steroid hormones in neuronal cells was investigated by studying oxidative stress-induced cell death caused by the neurotoxins amyloid beta protein, hydrogen peroxide and glutamate in the clonal mouse hippocampal cell line HT22. Preincubation of the cells with 10(-5) M 17-beta estradiol prior to addition of the neurotoxins prevented oxidative stress-induced cell damage and ultimately cell death, as detected with cell viability (MTT) and cell lysis (trypan blue exclusion/cell counting; propidium iodide staining) assays. At the DNA level, 17-beta estradiol blocked the DNA degradation caused by glutamate. Other steroid hormones, such as progesterone, aldosterone, corticosterone and the steroid precursor cholesterol, did not protect the cells. The neuronal protection afforded by 17-beta estradiol was estrogen receptor-independent. These data demonstrate a potent neuroprotective activity of the antioxidant 17-beta estradiol, which may have implications for the prevention and treatment of Alzheimer's disease. PMID- 7488137 TI - Establishment of a monoepoxide (leukotoxin and its isomer) producing system using a hydrogen peroxide-generating system. AB - We established an effective monoepoxide-generating system by combining cytochrome c (Cyt-c) with a hydrogen peroxide (H2O2)-generating system comprising hypoxanthine (HX), xanthine oxidase (XO) and superoxide dismutase (SOD; HX-XO-SOD Cyt-c system). Using this and the H2O2-Cyt-c system, we proved that monoepoxide production from linoleic acid was due to hydroxy radical formation by the reaction of Cyt-c with H2O2 and not to the formation of other active oxygen species. PMID- 7488138 TI - Different expression of the myotonin protein kinase gene in discrete areas of human brain. AB - We have examined the myotonin protein kinase (MT-PK) gene expression in different human brain areas. Four different spliced forms, comprising exons 13 and 14, were identified and characterized. One form (MYOT-A), lacking the entire exon 13, had not been detected in other studies and it is likely to be a brain-specific transcript. Different brain areas show a specific transcription pattern. These results suggest that MT-PK may have specialized functions in different areas of central nervous system. Alterations of this complex expression pattern could be responsible for the mental status impairment observed in myotonic dystrophy patients. PMID- 7488139 TI - The identification and partial characterisation of a novel inducible extracellular thermostable esterase from the archaeon Sulfolobus shibatae. AB - Extracellular esterases have so far only been reported in eubacteria, here we report the first identification and partial characterisation of a novel inducible extracellular esterase from the thermoacidophilic archaeon Sulfolobus shibatae. This esterase exhibits remarkable stability to both acid and heat. Esterase activity is induced by growth on a range of polyoxyethylenesorbitan (Tween) compounds as sole carbon source. Activity occurs over a wide temperature (25-99 degrees C) and pH (pH4.0-9.0) range and is optimal at 90 degrees C and pH6.0. It exhibits high thermal stability, with a half-life of 20 min at 120 degrees C, and shows a transient thermal activation of 60% at 90 degrees C. The thermal inactivation of function occurs by first order kinetics, and after 120 min incubation at 120 degrees C 50% of activity still remains. It is able to hydrolyse mono- and diglycerides, but is unable to hydrolyse the triglycerides olive oil and triolein, which is indicative of an esterase and not a lipase. PMID- 7488140 TI - Lesioned low density lipoprotein in atherosclerotic apolipoprotein E-deficient transgenic mice and in humans is oxidized and aggregated. AB - We analyzed lesioned LDL in both atherosclerotic humans and in the apo E deficient (E degree) mice and compared its characteristics to plasma LDL. Lesioned LDL, in comparison to plasma LDL, was minimally oxidized and aggregated. Upon incubation of E degree-aortic lesions with 125[I]-labeled LDL, a time dependent oxidation of the lipoprotein occurred as evident by a rapid and substantial elevation in LDL-associated TBARS from 0.2 to 10.3 and 14.5 nmoles of MDA equivalents/mg LDL protein after 2 and 24 hours of incubation, respectively. Only minimal LDL aggregates could be detected after 2 hours of incubation. Extensive LDL aggregation (15%), however, occurred after 24 h of incubation. Similar results were obtained on using human lesioned aortas. We conclude that both oxidation and aggregation of lesioned LDL could be the result of aortic lesioned-induced modification of the lipoprotein, and both of these modified forms of LDL can further contribute to the acceleration of the atherosclerotic process. PMID- 7488141 TI - Endothelin-1 expression by human hepatocellular carcinoma. AB - Plasma levels of endothelin-1 (ET-1) are markedly higher in patients with hepatocellular carcinoma (HCC) than in normal controls. In order to further investigate this, we evaluated ET-1 immunoreactivity and mRNA expression in human HCC tissue. 70% (14/20) of the tumor tissues immunostained positively with ET-1 antibody and a significant association was observed between immunostaining in cells lining the tumor feeding vessels and tumor vascularity as determined by hepatic angiography. Moreover, the neoplastic hepatocytes in the tumors also stained positively with ET-1 antibody. All of the HCC tissue samples which immunostained for ET-1 also expressed ET-1 mRNA, indicating that ET-1 is actively synthesized by the tumors. PMID- 7488142 TI - Differences in substrate specificity between Cdk2-cyclin A and Cdk2-cyclin E in vitro. AB - Cyclin-dependent kinase 2 (Cdk2), when bound to either cyclin A or cyclin E, recognizes the Ser/Thr-Pro-X-basic amino acid (motif A) as a phosphorylation site. In this study, we designed several peptides based on motif A and examined the substrate specificity of Cdk2-cyclin A and Cdk2-cyclin E using these peptides. Peptides containing a proline residue in the sequence Pro-X-Thr-Pro-X basic amino acid (motif B) had higher affinity for both Cdk2 complexes than peptides containing motif A. Furthermore, differences in substrate affinity between the two Cdk2 complexes were caused by a proline residue adjacent to or three positions before the threonine residue. Similarly, the presence of different basic amino acids in motif B also had different effects on affinity for each complex. We demonstrate the possibility that the substrate specificity of Cdk2 bound to cyclin might be regulated by the species of cyclin. PMID- 7488144 TI - Differential gene expression of vascular natriuretic peptide receptor subtype in artery and vein. AB - Although the vasorelaxation by natriuretic peptide (NP) is much less potent in the vein than in the artery, mechanism underlying the phenomenon remains unknown. Since NP receptor consists of three subtypes with different functions, we determined the mRNA level of each NP receptor subtype in the artery and vein by ribonuclease protection assay. In the aorta, NP-A receptor related to the biological action of NP was the predominant form. By contrast, NP-C receptor related mainly to the clearance of NP was the predominant form in the inferior vena cava: NP-C mRNA level was about two fold higher than in the aorta, while both NP-A and NP-B receptor mRNA levels were about half of that in the aorta. These results provide the molecular basis for the different biological response to NP in the artery and vein. Differential gene expression of NP receptor subtype could be an important determinant of the biological actions of NP. PMID- 7488143 TI - Molecular cloning and characterization of a new member of the RAC protein kinase family: association of the pleckstrin homology domain of three types of RAC protein kinase with protein kinase C subspecies and beta gamma subunits of G proteins. AB - cDNA clones encoding the third member of the RAC protein kinase family, termed RAC-PK gamma, were isolated from a rat brain cDNA library. The deduced amino acid sequence of RAC-PK gamma was highly related to those of previously identified family members, RAC-PK alpha and beta, that have a pleckstrin homology domain and a protein-serine/threonine kinase catalytic domain at the amino- and carboxyl terminal regions, respectively. Northern blot analysis indicated that RAC-PK gamma was expressed abundantly in brain and testis. Specific activities of RAC-PK alpha, beta, and gamma purified from transfected COS-7 cells were similar when measured by using myelin basic protein as a phosphate acceptor. Analysis using fusion proteins of glutathione S-transferase revealed that the pleckstrin homology domain of the three subtypes of RAC-PK associate with both protein kinase C subspecies and beta gamma subunits of G proteins. These results suggest that the pleckstrin homology domains of RAC protein kinase family could associate more than one protein to regulate the activity and/or intracellular distribution of this enzyme family by different ways. PMID- 7488146 TI - The directed evolution of radiation resistance in E. coli. AB - E. coli AB1157 (a wild-type, K-12 strain having no known defects in DNA repair capability) was irradiated daily with a very large X-ray dose to develop a series of strains unusually resistant to both X rays and ultraviolet (UV) photons. An understanding of how wild-type strains mutate and become more resistant should lead to a better understanding of DNA repair processes and their effects on radiation sensitivity. PMID- 7488145 TI - Lineage switch of a mouse pre-B cell line (SPGM-1) to macrophage-like cells after incubation with phorbol ester and calcium ionophore. AB - The mouse CD5 positive pre-B cell line SPGM-1 can be induced to switch its lineage commitment towards macrophage differentiation by treatment with a combination of phorbol ester and a calcium ionophore. When cultured with these reagents the pre-B cells ceased to proliferate and rapidly became adherent to plastic surfaces. This morphological change was accompanied by the loss of pre-B cell-specific surface markers, such as PB76 and most prominently the mu heavy chain of the immunoglobulin receptor complex. In addition, the mRNA of the surrogate light chain lambda 5 disappeared while the induction of lysozyme mRNA could be detected. Differentiated SPGM-1 cells phagocytosed latex beads and showed nonspecific esterase activity. The high efficiency and speed of differentiation in this cellular system makes SPGM-1 a highly suitable model for studying the phenomenon of lineage switching during hemopoesis. PMID- 7488147 TI - Lactate production from the rat hindlimb is increased after glucose administration and is suppressed by a selective amylin antagonist: evidence for action of endogenous amylin in skeletal muscle. AB - By serially measuring blood flow and venous-arterial lactate differences across the hindlimb of the fasted anesthetized rat, we examined (1) whether exogenous amylin increased muscle lactate production in vivo, (2) whether glucose administration increased muscle lactate production, and (3), by using the selective amylin antagonist AC187 to block endogenous peptide, whether amylin secreted in response to glucose could mediate muscle lactate production. Abdominal aortic flow was unchanged by any treatment. Hindlimb lactate production was increased by both 100 micrograms s.c. amylin (4.0 +/- 0.4 cf 2.6 +/- 0.3 mumol/min after saline, P < 0.05) and by infusion of 2mmol D-glucose (3.0 +/- 0.2 cf 2.3 +/- 0.2 mumole/hr after saline, P < 0.03). The increase in hindlimb lactate production was prevented by infusion of AC187 (mean post-treatment venoarterial delta-lactate 140 +/- 11 microM; n.s. vs saline-treated delta lactate 154 +/- 10 microM; P < 0.05 vs glucose-treated delta-lactate 201 +/- 14 microM). These findings are consistent with endogenous amylin secreted in response to a glucose challenge having acted at skeletal muscle to release lactate. PMID- 7488148 TI - Regulation of the human serotonin transporter by interleukin-1 beta. AB - The serotonin transporter expressed in brain, platelets, and placenta is a primary target for antidepressants, cocaine, and amphetamines. Here we report that interleukin-1 beta is a potent regulator of the expression of this transporter. The activity of the serotonin transporter in human JAR choriocarcinoma cells is stimulated by this cytokine and the stimulation is accompanied by an increase in the steady state levels of the transporter mRNAs and in the transporter density. The increase in mRNA levels is effectively blocked by actinomycin D. The stimulatory effect of interleukin-1 beta is not associated with any change in the cellular levels of cAMP, indicating involvement of a cAMP-independent pathway. Even though cholera toxin, which is known to increase cAMP levels, also upregulates the serotonin transporter in JAR cells, the effects of interleukin-1 beta and cholera toxin on the transporter activity are additive. This constitutes the first report on the identification of a physiologically occurring hormone/cytokine which upregulates the expression of the human serotonin transporter gene. PMID- 7488149 TI - Cupric ion blocks NF kappa B activation through inhibiting the signal-induced phosphorylation of I kappa B alpha. AB - A transcription factor NF kappa B, which regulates expression of various cellular genes involved in immune responses and viral genes including HIV, is sequestered in the cytoplasm as a complex with an inhibitory protein I kappa B. Various extracellular signals induce phosphorylation and rapid degradation of I kappa B alpha to release NF kappa B. Cu2+ was found to inhibit the activation of NF kappa B induced by TNF-alpha, TPA, or H2O2. Deoxycholate treatment of the cytoplasmic extract prepared from cells stimulated by TNF-alpha in the presence of Cu2+ resulted in the release of NF kappa B from I kappa B alpha, indicating that Cu2+ interferes with the dissociation of the NF kappa B-I kappa B complex. Neither phosphorylation nor degradation of I kappa B alpha was observed upon TNF-alpha stimulation in the presence of Cu2+. These results indicate that Cu2+ inhibits the release of NF kappa B by blockade of a signal leading to the phosphorylation of I kappa B alpha. PMID- 7488150 TI - Midkine is a heat and acid stable polypeptide capable of enhancing plasminogen activator activity and neurite outgrowth extension. AB - We studied about the physicochemical stability of a novel heparin-binding growth/differentiation factor, midkine (MK). It was found that synthetic human MK was heat and acid stable. Neither incubation at 80 degrees C for 90 sec nor treatment at low pH affected the elution profile of MK molecule on the high performance liquid chromatography. This physicochemical stability was maintained in the biological activities of MK to enhance plasminogen activator (PA) activity in bovine endothelial cells as well as to promote neurite outgrowth of rat brain cells. A similar stability was observed both with recombinant murine MK and with its homologous protein, recombinant human pleiotrophin. Comparison of physicochemical stability with other several growth factors suggested that MK/pleiotrophin was a unique family of heat and acid stable polypeptides capable of enhancing PA activity and neurite outgrowth. PMID- 7488152 TI - Vanadate and bafilomycin A1 are potent inhibitors of the ATPase activity of the reconstituted bacterial ATP-binding cassette transporter for maltose (MalFGK2). AB - Vanadate and Bafilomycin A1 were shown to inhibit the ATPase activity of the reconstituted binding protein-dependent ATP-binding Cassette (ABC) transporter for maltose (MalFGK2) of Salmonella typhimurium in the micromolar range. This is in sharp contrast to the recent finding that the isolated ATPase subunit MalK was insensitive to both compounds. Our data provide the first experimental evidence for the view that functional coupling of the ATPase domain of an ABC transporter to the membrane-integral domains is crucial for conferring sensitivity to vanadate and bafilomycin A1. Possible consequences for the mode of action of ABC transport proteins are discussed. PMID- 7488153 TI - High level expression of the IL-1 receptor related T1 receptor in insect cells. AB - Baculovirus-dependent expression of membrane-associated and secreted, heavily modified proteins in insect cells often results in low yields. To optimize expression of T1, a heavily glycosylated receptor related to Interleukin-1 receptor type I, deletion mutants comprising either the heavily glycosylated extracellular domain or the isolated transmembrane and cytoplasmic portions of the T1 receptor were expressed from recombinant baculoviruses. As shown here, the use of a Baculovirus-derived leader sequence (GP67) in combination with an insect cell line with increased secretory capacity resulted in yields of several mg per liter (10(9) cells) of culture of both purified secretory T1 glycoprotein and membrane-associated T1 receptor. PMID- 7488151 TI - NMDA receptor subunits epsilon 1 (NR2A) and epsilon 2 (NR2B) are substrates for Fyn in the postsynaptic density fraction isolated from the rat brain. AB - Fyn, protein tyrosine kinase, and its substrates were highly concentrated in the postsynaptic density (PSD) fraction prepared from the rat forebrain. There were a number of Fyn substrates unique to the PSD fraction. One of the major substrates in the PSD fraction was found to be a concanavalin A-binding glycoprotein, PSD gp180, which is the N-methyl-D-aspartate (NMDA) receptor subunit epsilon 2 (NR2B). Western blotting and immunoprecipitation supported the phosphorylation of epsilon 2 by Fyn. NMDA receptor subunit epsilon 1 (NR2A) was also a substrate for Fyn. These results suggest that Fyn is involved in the modulation of synaptic efficacy through the phosphorylation of synapse-specific substrates such as the NMDA receptor/channel. PMID- 7488154 TI - Drug-stimulated ATPase activity of a deletion mutant of the human multidrug resistance protein (MDR1). AB - The baculovirus-insect cell system has been used for the functional expression of the human multidrug resistance protein (MDR1) and a mutant MDR1 variant lacking a twenty amino acid segment from the first extracellular loop (delta aa78-97 MDR1). Both MDR1 proteins were found to be correctly inserted into the insect cell membrane as indicated by their interaction with MRK 16 antibody. The removal of the 78-97 segment from the first extracellular loop dramatically altered drug stimulated ATPase activity. Rhodamine 123 or vinblastine were not able to stimulate the mutant protein and Calcein AM had also little effect. In contrast, verapamil increased the ATPase activity of the mutant almost to the same maximal level as that of the wild type. However, the verapamil concentration needed for the half maximal stimulation of the ATPase activity was found to be about hundred times higher than that for the wild type MDR1. These results indicate that a partial deletion of an extracellular loop modulates the affinity of MDR1 for its transportable substrates in a variable fashion. PMID- 7488155 TI - The use of RT-PCR to distinguish between plasmid MnSOD transcripts and endogenous MnSOD mRNA. AB - We report here a convenient RT-PCR method to distinguish plasmid human MnSOD cDNA transcripts from the endogenous MnSOD gene products without engineering the cDNA insert. When a specific antisense primer for the carrier vector sequence was paired with a sense primer for the human MnSOD cDNA in RT-PCR analysis, a unique amplicon with the expected size was generated in MnSOD cDNA transfected cells but not in the wild type or vector control cells. The same primers were also used in genomic DNA-PCR to demonstrate genomic incorporation of cDNA in stably transfected cells. This method is convenient and specific in determining exogenous cDNA incorporation and expression in transfectants especially when transcripts of cDNA are difficult to separate from the endogenous mRNA by other methods. PMID- 7488156 TI - Novel phosphorylation at a mitotic site, serine 75, in human pp60c-src from unsynchronized human tumor cells having a spherical morphology. AB - Endogenous pp60c-src from Y79 unsynchronized human retinoblastoma cells is phosphorylated at an additional N-terminal serine residue relative to unsynchronized fibroblast pp60c-src. We confirmed that the novel phosphorylation site is Ser 75, which is the same as that phosphorylated in overexpressed human pp60c-src during NIH3T3 cell mitosis. We also showed that the Ser 75 phosphorylation pattern correlates with cell rounding in 14 various unsynchronized human tumor cell lines. Especially, pp60c-src from Lu135 having a spherical morphology similar to that of Y79 is phosphorylated as high as Y79 pp60c-src. Mitotic spherical cells of the epithelial-like HepG2 express pp60c-src phosphorylated on Ser 75. On the other hand, Y79 pp60c-src is phosphorylated on Ser 75 throughout the cell cycle. These data suggest that this phosphorylation may be important in spherical cell morphology. PMID- 7488158 TI - Extracellular nucleotides increase cytosolic free calcium by activating P2u receptors in single human gastric mucous cells. AB - In single human gastric mucous cells extracellular ATP, ATP-gamma-S, ADP and UTP at micromolar concentrations caused a biphasic increase in [Ca2+]i. Consistent with a P2u-receptor the rank-order of potency was ATP > or = UTP > ATP-gamma-S > ADP, and cross-desensitization of the Ca2+ responses occurred between ATP and UTP. The initial transient peak in [Ca2+]i was resistant to extracellular Ca2+ depletion which demonstrates mobilization of internal Ca2+. By contrast, the sustained plateau phase required influx of external Ca2+. Ca2+ influx occurs most likely through a capacitative Ca2+ entry mechanism, which was shown to exist in these cells by experiments performed with thapsigargin. Thus, extracellular ATP and UTP activate a common P2u-receptor most likely coupled to the Ca(2+) phosphatidylinositol signalling cascade. Extracellular nucleotides from various sources might be an important factor in the regulation of human gastric mucous cells. PMID- 7488159 TI - An Ael allele-specific nucleotide insertion at the blood group ABO locus and its detection using a sequence-specific polymerase chain reaction. AB - Genomic DNA from each of four Acl individuals (genotypes AO1, AO1var, AO2) and one AclB individual was used as a template for amplifying exons 6 and 7 of the ABO genes, which were subsequently sequenced. In all the Ael alleles a single nucleotide insertion, compared to the A consensus sequence, was observed that would alter the amino acid sequence of the glycosyltransferase immediately after its postulated nucleotide sugar binding site and furthermore extend the translated protein by 37 amino acids (16 more than the A2 enzyme). A sequence specific primer PCR assay was developed to detect the nucleotide insertion. It was possible to differentiate all 20 serologically defined Acl/AclB individuals available from 145 blood donors with normal ABO phenotypes and genotypes and 26 individuals with various A subgroups other than A1, A2 and Acl. This mutation explains the Acl phenotype and forms the basis of a method for detecting the Ael allele. PMID- 7488157 TI - Fine mapping of MEP1A, the gene encoding the alpha subunit of the metalloendopeptidase meprin, to human chromosome 6P21. AB - Meprins are kidney and intestinal proteases encoded by two distinct genes, MEP1A and MEP1B. MEP1A was previously mapped to human chromosome 6p, by the use of radiation and somatic cell hybrids, in the region containing the gene for autosomal recessive polycystic kidney disease (ARPKD). We now report the fine mapping of MEP1A using yeast artificial chromosome clones, and linkage analysis of ARPKD families. The results from both physical and genetic mapping exclude MEP1A as a candidate for ARPKD. These studies place MEP1A in a region more telomeric to 6p12 and closer to the HLA loci than previously reported. More specifically, MEP1A is localized between loci D6S272 and D6S282, close to D6S452, on human chromosome 6p21.2-p21.1. The more precise location of MEP1A will facilitate genetic studies of this locus and clarify the relation of this gene to others. PMID- 7488160 TI - Mutation of the carboxy terminal zinc finger of E. coli isoleucyl-tRNA synthetase alters zinc binding and aminoacylation activity. AB - Escherichia coli isoleucyl-tRNA synthetase has been shown to contain two enzyme bound zinc atoms per polypeptide chain. To investigate the structural and functional significance of the C-terminal enzyme-bound zinc, mutagenesis was used to alter Cys 922 to Ser [IleRS(C922S)] and to replace Cys 922 through Ala 939 with a 33 amino acid peptide unable to bind zinc (AIleRS). Both IleRS(C922S) and AIleRS were found to contain only a single enzyme-bound zinc per polypeptide chain. Substitution of Co2+ for Zn2+ in IleRS(C922S) gave a visible spectrum characteristic of that expected for a single tetrahedrally coordinated enzyme bound Co2+ atom. Little or no effect on the Km values for ATP or Ile and only a 5 fold reduction of the (kcat/Km)Ile was observed for IleRS(C922S) and AIleRS in the isoleucine-dependent ATP-pyrophosphate exchange reaction. In the tRNA dependent aminoacylation reaction, Km values for tRNA(Ile) were only slightly affected in the mutant proteins. However, kcat/Km values were decreased approximately 200 and 2500 fold for IleRS(C922S) and AIleRS, respectively. These results suggest that both the C-terminal enzyme-bound zinc and the C-terminal peptide play important roles in aminoacylation of tRNA(Ile). PMID- 7488162 TI - The bone-specific estrogen centchroman inhibits osteoclastic bone resorption in vitro. AB - There is considerable interest in identifying bone-specific estrogen-like compounds with beneficial activities on bone and the cardiovascular system, but lacking side effects on the reproductive system. Two such compounds are currently under clinical investigation -raloxifene (Lilly) and centchroman (Novo-Nordisk). There is evidence suggesting that 17 beta-estradiol can inhibit osteoclastic bone resorption although this is somewhat controversial. Therefore, we examined the effect of centchroman and raloxifene, as well as 17 beta-estradiol, in the in vitro bone slice assay, where the direct effect of compounds on osteoclast activity can be assessed. Centchroman (0.001 - 1 microM) dose-dependently inhibited osteoclastic bone resorption up to 70% at 1 microM (p = 0.007) with an IC50 = 0.1 microM, while in contrast, raloxifene had no significant effect on bone resorption over the same dose range. 17 beta-estradiol (0.0001 - 1 microM) had a modest but significant inhibitory effect on resorption (40%, p < 0.05) at 1 microM, but no effect at lower physiological/therapeutic concentrations. Centchroman (1 microM) inhibited osteoclast cytoplasmic spreading by 32%, while raloxifene and 17 beta-estradiol were without effect. These results show that centchroman at therapeutic concentrations (ED50 approximately 1 mg/kg in animal models) is a potent inhibitor of osteoclastic bone resorption in vitro, suggesting that bone-specific estrogen-like molecules may have different mechanisms of action. PMID- 7488161 TI - Radiolabeling of catalytic subunits of PI 3-kinases with 17 beta-hydroxy-16 alpha(-)[125I]iodowortmannin: identification of the G beta gamma-sensitive isoform as a complex composed of 46-kDa and 100-kDa subunits. AB - A fungal metabolite, wortmannin, is a potent inhibitor of phosphatidylinositol (PI) 3-kinases. In the present study, we prepared a radiolabeled derivative of wortmannin, 17 beta-hydroxy-16 alpha(-)[125I]iodowortmannin. The compound bound tightly to a 110-kDa subunit in the previously identified isoform of PI 3-kinase (p85/p110), and also to a 100-kDa peptide in a partially purified preparation of another isoform of PI 3-kinase whose activity was markedly stimulated by the beta gamma subunits of GTP-binding proteins (G beta gamma). The binding to both peptides was inhibited by non-radiolabeled wortmannin and also by LY294002, another inhibitor of PI 3-kinases. An antibody against p85 recognized a 46-kDa peptide in the G beta gamma-sensitive isozyme and precipitated the 100-kDa peptide specifically labeled with 17 beta-hydroxy-16 alpha( )[125I]iodowortmannin. These results suggested that the newly found isozyme was a complex composed of 46-kDa and 100-kDa peptides. PMID- 7488163 TI - Effect of protein kinase and phosphatase inhibitors on expression of hypoxia inducible factor 1. AB - Hypoxia-inducible factor 1 (HIF-1) is a heterodimeric bHLH-PAS protein essential for erythropoietin gene transcription in hypoxic cells. Here we show that both 2 aminopurine and sodium fluoride, inhibitors of serine/threonine kinases and phosphatases, respectively, interfered with the hypoxic induction of HIF-1 DNA binding activity and expression of HIF-1 alpha and HIF-1 beta(ARNT) subunits. Genistein, an inhibitor of tyrosine kinases, completely blocked the synthesis of both HIF-1 subunits as well as HIF-1 DNA-binding activity. Sodium orthovanadate, an inhibitor of tyrosine phosphatases increased the basal level of HIF-1 proteins and HIF-1 activity. These data suggest that protein phosphorylation events play an important role in the hypoxia signal-transduction pathway that leads to synthesis of HIF-1 alpha and HIF-1 beta proteins and the induction of HIF-1 DNA binding activity. PMID- 7488164 TI - Oxygen regulation of the cytochrome c oxidase subunit VI gene, COX6, in Saccharomyces cerevisiae. AB - Cytochrome c oxidase (CcO) in Saccharomyces cerevisiae is subject to intricate physiological control. Growth phase, carbon source, and oxygen level are three well recognized modulators of CcO expression. We focused on the subunit VI encoding gene, COX6, and detected unexpectedly complex oxygen regulation. We found that COX6 transcription possessed a critical threshold oxygen regulation between 0 and 2%. COX6 transcription was superinduced by elevated oxygen level up to 45%; however, superinduction was lost at 60% oxygen and above. The COX6 upstream activation region, UAS6, contains both glucose and heme responsive regions, and COX6 oxygen regulation was transduced through UAS6 by heme, as has been described for other oxygen regulated genes in yeast. We found that binding of the UAS6-domain 1 protein, BAF1, was unaltered by oxygen regulation. Nor were the alternative BAF1 complexes observed by growth in different glucose conditions formed by growth at different oxygen levels. PMID- 7488166 TI - Genotoxicity induced by furocoumarin hydroperoxides in mammalian cells upon UVA irradiation. AB - The mutagenicity and micronucleus induction by the furocoumarin hydroperoxides 1a and 2a and, for comparison, their corresponding alcohols 1b and 2b were investigated in L5178Y tk+/- mouse lymphoma cells and AS52 Chinese hamster ovary cells. The furocoumarin hydroperoxide 1a enhanced the micronucleus frequency in L5178Y tk+/- mouse lymphoma cells significantly, while its alcohol 1b exhibited a rather moderate effect. In contrast, both the furocoumarin hydroperoxide 2a and its alcohol 2b induced high frequencies of micronuclei. Only the furocoumarin hydroperoxide 1a, but not its alcohol 1b, was mutagenic in both L5178Y tk+/- and AS52 cells. On the other hand, the furocoumarin hydroperoxide 2a and its alcohol 2b were mutagenic. For all furocoumarin derivatives 1a,b and 2a,b no mutagenicity and genotoxicity was observed in the absence of UVA light. The multifunctional furocoumarin hydroperoxides may serve as potential photochemotherapeutic agents. PMID- 7488165 TI - A new perspective on the inhibitory role of nitric oxide in sympathetic neurotransmission. AB - By using, as a model of sympathetic neurons, immortalized rat pheochromocytoma (PC12) cells differentiated by nerve growth factor (NGF), the effect of nitric oxide on sympathetic neurotransmission was examined. The NO donor sodium nitroprusside (SNP; 10(-4)-3 x 10(-4) M) caused an apparent inhibition of dopamine release from PC12 cells, as measured by HPLC. Studies, in the absence of cells, involving the incubation of dopamine (20 ng/ml) or norepinephrine (15 ng/ml) with SNP (10(-6)-3 x 10(-4) M) or authentic NO (6 x 10(-6)-3 x 10(-5) M) revealed a similar reduction in the detection of the catecholamines. In addition, absorption spectroscopy studies showed dopamine and norepinephrine to be oxidized by NO resulting in the formation of their respective quinone products. These observations, coupled with the finding that the ability of dopamine to raise cAMP levels within PC12 cells was reduced after incubation with SNP, reveal that NO inhibits the biological activity rather than the release of catecholamines. PMID- 7488167 TI - The role of iron in beta amyloid toxicity. AB - There is evidence that oxidative damage plays a causative role in Alzheimer's disease and amyloid beta protein (A beta) toxicity. Iron is frequently a potent facilitator of free radical production due to its ability to mediate the conversion of H2O2 to hydroxyl radicals via the Fenton reaction or by virtue of hypervalent iron compounds. It is shown here that iron facilitates A beta toxicity to cultured cells. PMID- 7488168 TI - Is polyamine decrease a common feature of apoptosis? Evidence from gamma rays- and heat shock-induced cell death. AB - Here we report that in rat thymocytes undergoing apoptosis upon two different stimuli, such as heat shock treatment and gamma irradiation, an early mRNA accumulation of ornithine decarboxylase (ODC)--the rate-limiting enzyme of polyamine biosynthesis--was followed by a very marked increase in ODC activity (28-40 and 6-8-fold, respectively). However, polyamine levels started to decrease before the appearance of DNA laddering, being putrescine and spermidine strongly diminished (8-12 hs), and spermine even depleted (12 hs). Taken together with our previous data on another model of apoptosis, i.e., glucocorticoid-induced cell death (Desiderio et al., Cell Growth Differ. 6: 505-513, 1995), these results suggest that an imbalance of polyamine metabolism, i.e., a strong activation of ODC and a paradoxical decrease of the intracellular polyamine content, might be a general feature of the apoptotic process. PMID- 7488169 TI - Stimulation of DNA polymerase gamma activity by proliferating cell nuclear antigen. AB - DNA polymerase was partially purified from mitochondrial extracts of rat liver by phosphocellulose, DEAE-cellulose, heparin-Sepharose CL-6B and DNA-agarose column chromatography. By these purification steps, DNA polymerase and proliferating cell nuclear antigen (PCNA) were completely separated at the step of heparin Sepharose CL-6B column chromatography. The isolated DNA polymerase was inhibited by ddTTP, but not by aphidicolin. The enzyme sedimented at about 8 S on 5-20% analytical sucrose density gradient centrifugation. These data showed that the DNA polymerase isolated from mitochondria is gamma in type. After the separation of DNA polymerase gamma and PCNA, the two fractions were remixed and DNA polymerase gamma activity was measured. DNA polymerase gamma activity was stimulated about three-fold or more in the presence of the PCNA fraction. This stimulation was inhibited by the addition of anti-PCNA rabbit IgG2a. In addition, highly purified human recombinant PCNA stimulated the DNA polymerase gamma activity. These results indicate that DNA polymerase gamma, like DNA polymerase delta, is activated by PCNA. PMID- 7488170 TI - Classic steroid 11 beta-hydroxylase deficiency caused by a C-->G transversion in exon 7 of CYP11B1. AB - Steroid 11 beta-hydroxylase deficiency (11 beta OHD) is derived from mutations in the P45011 beta gene (CYP11B1) and inherited in an autosomal recessive manner. In the present study, we have performed a molecular genetic analysis of CYP11B1 in a Japanese patient clinically diagnosed as classic 11 beta OHD. Nucleotide sequencing of the PCR-amplified exons from the patient's genomic DNA reveals a unique C-->G transversion that converts codon 384 CGA (arginine) to GGA (glycine) in exon 7. Restriction fragment length polymorphism (RFLP) data demonstrate that the patient is homozygous for this mutation. When the full-length cDNA corresponding to CYP11B1 of the patient is transfected into COS-7 cells, no steroid 11 beta-hydroxylase activity is detectable in mitochondria of the cells. These results indicate that this point mutation completely abolishes P45011 beta activity and causes the classic 11 beta OHD. PMID- 7488171 TI - A SP1 binding site in the GC-rich region is essential for a core promoter activity of the human endothelial nitric oxide synthase gene. AB - Endothelial nitric oxide synthase (eNOS) is an important oxygenase which catalyzes the conversion of L-arginine to L-citrulline to form nitric oxide (NO), a potent important factor for vasodilation and inhibition of platelet aggregation. We have analyzed characteristics of the promoter region of the human eNOS gene using the transient expression in human endothelial cells of CAT constructs with a series of 5'-deletion mutants. The 5'-flanking region between 116 and -98, which contains a putative consensus sequence for binding of transcription factor Sp1, is essential to direct a basal promoter activity. Gel mobility shift analysis involving anti-Sp1 antibody and competitor DNAs disrupted at the binding site for Sp1 reveals that Sp1 or its closely related protein(s) binds to the consensus sequence located between -104 and -96. These results indicate that the Sp1 site is essential for a core promoter activity of the human eNOS gene. PMID- 7488172 TI - Protective effect of coenzyme Q10 on cultured skeletal muscle cell injury induced by continuous electric field stimulation. AB - The protective effect of coenzyme Q10 (CoQ10) on continuous electric field stimulation-induced muscular injury was investigated in cultured cells established from neonatal rat femoral muscles. After cultivation for 9 days, skeletal muscle cells contracted and relaxed rhythmically for 4 hr in response to continuous electric field stimulation (power, 5 V; duration, 5 msec; amplitude, 3 Hz). After the onset of the stimulation, lactate and lactate dehydrogenase (LDH) release and intracellular Ca2+ contents ([Ca2+]i) at relaxation increased gradually. In contrast, the intracellular ATP contents decreased. The addition of 5 microM CoQ10, but not alpha-tocopherol and radical scavengers, to the culture medium protected the cells against these biochemical changes after the stimulation. Verapamil, an inhibitor of Ca2+ channels, also attenuated the increase in [Ca2+]i at relaxation and LDH. These results suggested that one of the causal mechanisms of muscular injury is an increase in [Ca2+]i due to the excess entry of extracellular Ca2+, and that CoQ10 can protect skeletal muscle cells against such undesirable biochemical changes. PMID- 7488173 TI - Expression of fungal Mn peroxidase in E. coli and refolding to yield active enzyme. AB - The cDNA encoding Mn peroxidase isozyme H4 from Phanerochaete chrysosporium was expressed in Escherichia coli. The portion of the cDNA encoding the enzyme's signal peptide, not found in the processed holoenzyme, was deleted from the cDNA. The polypeptide was produced as inactive inclusion bodies that could be solubilized in 8 M urea and the reducing agent dithiothreitol. Reconstitution of activity was accomplished by diluting the urea concentration to 2M in the presence of hemin, calcium, and oxidized glutathione. All of the additives were required for recovery of activity. The activity of the recombinant enzyme was dependent on both Mn2+ and H2O2. PMID- 7488174 TI - A template for generation and comparison of three-dimensional selectin models. AB - We have complemented multiple sequence alignments of the lectin domains of the selectins with an analysis of structurally invariant regions in X-ray structures of the mannose-binding protein (MBP) and E-selectin. The analysis shows that regions of structural conservation between MBP and E-selectin extend beyond regions of rigorous sequence conservation within the selectin family and suggests that reliable three-dimensional models of selectins from different species can be generated by modification of only a few backbone segments in E-selectin. A model of the L-selectin lectin domain is built and discussed with regard to observed differences in selectin specificity. PMID- 7488175 TI - Production of a form-specific, inhibitory antibody against rat cytochrome P450 2B1 using a synthetic peptide antigen against a putative substrate binding site. AB - Rat cytochrome P450 2B1 antipeptide antibodies were produced by immunizing rabbits with a synthetic peptide antigen. The anti-CYP2B1 IgG obtained did not cross-react with CYP2B2, which has 97% identity in primary sequence of CYP2B1. This result demonstrates that a difference of 2 amino acid residues among 12 is sufficient to produce a form-specific antibody. The CYP2B1 antipeptide IgG inhibited pentoxyresorufin O-dealkylase activity of microsomes obtained from phenobarbital-treated rats in a dose-dependent manner, whereas it did not inhibit ethoxyresorufin O-deethylase activity of microsomes obtained from 3 methylcholanthrene-treated rats. These results suggest that the selected amino acid sequence, which coincides with one of the substrate binding sites of Pseudomonas putida CYP101A (P450cam) and one of the putative substrate binding sites of CYP2B2, is located on the surface of the CYP2B1 molecule, as opposed to inside the molecule or in the lipid bilayer of microsomes. PMID- 7488176 TI - Expression of protein kinase C isozymes that are required for chondrogenesis of chick limb bud mesenchymal cells. AB - Protein kinase C (PKC) has been suggested to be involved in the chondrogenesis of chick limb bud mesenchymal cells. This study examined the expression and the role of PKC isozymes in chondrogenesis. Multiple PKC isozymes such as conventional PKC (cPKC alpha and gamma), new PKC (nPKC epsilon), and atypical PKC (aPKC zeta, lambda, and tau) were expressed in chondroblasts but cPKC beta and nPKC delta were not detected. The amounts of expressed cPKC and nPKC isozymes, namely cPKC alpha and gamma and nPKC epsilon, were increased as chondrogenesis proceeds while the level of aPKC isozymes was not changed. Treatment of cells with specific PKC inhibitors blocked chondrogenesis. Prolonged exposure of cells to phorbol ester which down regulates both cPKC and nPKC also blocked chondrogenic differentiation. The inhibition of chondrogenesis was the most effective when PKC activity was blocked at the early stage of chondrogenesis (i.e., for the first 24 hours of micromass culture). Down regulation of PKC blocked both proliferation of cells and synthesis of sulfated proteoglycans, indicating that expression of cPKC and nPKC is required at early stage of chondrogenesis. PMID- 7488177 TI - Induction of heat-shock proteins and accumulation of trehalose by TPN in Saccharomyces cerevisiae. AB - TPN [Tetrachloroisophthalonitrile], a kind of disinfectant, affected growth in Saccharomyces cerevisiae. Exposed to TPN under no lethal conditions, 70-, 90-kDa protein and hsp104 were induced. Each of them was not uniformly induced; namely, the 70-kDa protein was more sensitive to TPN among other proteins. Trehalose was also accumulated depending on the concentration. The degree of thermotolerance in yeast cells pretreated with 1.0 mg/l TPN was about 100-fold greater than in the control. Under this condition, TPN-inducible proteins were synthesized but trehalose was not accumulated. Although TPN-inducible proteins and trehalose were definitely synthesized with 10.0 or 100.0mg/l TPN treatment, thermotolerance was not acquired. PMID- 7488178 TI - Characterization of the subunit isoforms of duck salt gland Na/K adenosine triphosphatase. AB - The N-terminal sequences of the alpha and beta subunits from the Na/K-ATPase of duck salt gland have been determined by automated Edman degradation chemistry. These sequences were compared to sequences previously reported for Na/K-ATPase subunits from other sources in order to determine the subunit isoform composition of the salt gland enzyme. The comparisons indicate that the duck salt gland enzyme is composed of an alpha-1 subunit and a beta-1 subunit. This subunit isoform composition is consistent with the involvement of this enzyme in sodium excretion as Na/K-ATPases in other tissues involved in sodium excretion also have this subunit isoform composition. PMID- 7488179 TI - Topography of the 27- and 31-kDa electron transport proteins in the onion root plasma membrane. AB - Plasma membranes purified from onion roots contain two distinct NAD(P)H dehydrogenases of 27 and 31 kDa that differ in their physicochemical properties, substrate specificities and inhibitors sensitivities. The 27-kDa enzyme used both NADH and NADPH as electron donors. The 31-kDa enzyme was fully specific for NADH and accounted for the bulk of NADH-ferricyanide oxidoreductase. We have used NADPH- and NADH-ferricyanide oxidoreductase activities as markers for investigating the orientation of the 27- and 31-kDa enzymes at the plasma membrane, respectively. These activities were assayed in right-side-out vesicles isolated by two-phase partition, inside-out vesicles obtained by treatment with the detergent Brij 58 and membranes permeabilized with Triton X-100. Upon addition of Brij 58 to right-side-out plasma membrane vesicles, both NADPH- and NADH-ferricyanide oxidoreductases were activated to the same degree as the plasma membrane H(+)-ATPase. Redox activities were similar when measured in the presence of either Brij 58 or Triton X-100. Our results demonstrate that both enzymes expose their catalytic sites toward the cytoplasmic side of the plasma membrane. PMID- 7488180 TI - Sexually dimorphic expression of pituitary growth hormone-releasing factor receptor in the rat. AB - Secretion of growth hormone (GH) exhibits marked sexual dimorphism in the rat. To examine the underlying mechanism that involves hypothalamic GH-releasing factor (GRF), we determined pituitary GRF receptor mRNA levels in male and female rats and compared their in vitro abilities to release GRF, an endogenous ligand for GRF receptor. Female rats expressed GRF receptor mRNA at a level of only 15% (P < 0.001) of that of male rats. Female rats also showed a 33% lower (P < 0.01) ability to release GRF than male rats. These results indicate that the GRF secretion and action system of female rats is characterized by the combined reduction in GRF receptor expression and GRF-releasing capacity compared with that of male rats. This could explain the in vivo finding that spontaneous, GRF triggered GH pulses are of much lower amplitude in the female than in the male rat. PMID- 7488181 TI - Plant growth hormone kinetin delays ageing, prolongs the lifespan and slows down development of the fruitfly Zaprionus paravittiger. AB - A cytokinin plant growth hormone kinetin (Kn) retards senescence in plants and delays ageing in human cells in culture. We have now observed that Kn also slows down ageing and prolongs the lifespan of the fruitfly Zaprionus paravittiger when these insects are fed with 25-125 ppm Kn added in their diet medium. In addition, 25 ppm Kn also slows down the larval and the pupal stages of the developing insects. However, the anti-ageing effects of Kn are not merely due to the slowing down of development, but are mainly due to a reduction in the age-specific death rates throughout the adult lifespan. PMID- 7488182 TI - The GGNG peptides: novel myoactive peptides isolated from the gut and the whole body of the earthworms. AB - Three novel bioactive peptides, which were structurally related to each other, were purified and chemically identified from two species of the earthworms, Eisenia foetida and Pheretima vittata. One peptide was isolated from the gut tissue of E. foetida, and the other two were purified from the whole body of E. foetida and P. vittata, respectively. These peptides consisted of 17 or 18 amino acid residues and were named GGNG peptides after their unique, common structure of the C-termini. These GGNG peptides augmented both tension and frequency of spontaneous contractions of isolated gut preparations of E. foetida, and also elicited contractions of other annelidan tissues such as the polychaete esophagus and the leech vagina. However, they showed no activity on molluscan or arthropodan tissues, suggesting that GGNG peptides may be peculiar to annelids. No peptides homologous to GGNG peptides have been known so far in any living organisms. PMID- 7488183 TI - Peroxinectin, a novel cell adhesion protein from crayfish blood. AB - From blood cells of the crayfish Pacifastacus leniusculus a 76-kDa protein that mediated attachment and spreading of the crayfish blood cells was purified. The cDNA for this cell adhesion protein was isolated, cloned, and sequenced. The deduced protein sequence was significantly similar to one family of peroxidases, e.g., myeloperoxidase. Consistently, the 76-kDa protein, for which we propose the name peroxinectin, had peroxidase activity. A synthetic peptide derived from the peroxinectin sequence containing Lys-Gly-Asp mimicked the cellular activity of the intact protein, implicating this sequence as the cell-binding site. Peroxinectin is the first cell adhesion molecule cloned from invertebrate blood and, to our knowledge, the first protein from any organism that combines being a cell adhesion ligand and a peroxidase. PMID- 7488184 TI - cDNA sequence analysis and expression of alpha-bungarotoxin from Taiwan banded krait (Bungarus multicinctus). AB - The cDNA encoding alpha-bungarotoxin (alpha-BuTx) was constructed from the cellular RNA isolated from the venom glands of Bungarus multicintus by RT-PCR. The primers deduced from the N-, C-terminal amino acid residues and the conserved middle segment of the sequence, i.e., R25KMWC29 in alpha-BuTx could specifically amplify cDNAs encoding amino acid residues from 1 to 29 and 25 to 74 of alpha BuTx. The complete cDNA was obtained by linking sufficient amounts of the two cDNA fragments by PCR. The amino acid sequence translated from the alpha-BuTx cDNA sequence was identical to that of native toxin. The PCR product was then cloned into pGEX vector and transformed in DH5 E. coli strain. The fused protein expressed in host strain exhibited the same antigenicity as native alpha-BuTx. This communication is the first report for the cloning and expression of a long alpha-neurotoxin from the venom glands. PMID- 7488185 TI - Properties of D-hydantoinase from Agrobacterium tumefaciens and its use for the preparation of N-carbamyl D-amino acids. AB - Agrobacterium tumefaciens strain 47C expresses an inducible D-hydantoinase that catalyzes the formation of optically pure N-carbamyl D-amino acids from racemic hydantoin precursors. The D-Hydantoinase was shown to be active and stable at elevated temperatures and pH values, thus affording favorable bioreaction conditions that result in the racemization of DL-hydantoins to the utilizable D isomer. The enzyme demonstrated optimal reaction kinetics at pH 10 and 70 degrees C, was not activated by metal ions, and exhibited a distinctive substrate specificity. A. tumefacins hydantoinase was most active on 5,6-dihydrouracil and DL-5-methylhydantoin with only slight activity on DL-benzylhydantoin. Extracts or whole cells of A. tumefaciens were used as biocatalyst to mediate the stereospecific conversion of DL-phenylhydantoin or DL-5-methylhydantoin to the respective N-carbamyl D-amino acids. In addition, immobilized cell systems were shown to be useful for biocatalyst reuse. PMID- 7488186 TI - Mouse rhodanese gene (Tst): cDNA cloning, sequencing, and recombinant protein expression. AB - Rhodanese (thiosulfate sulfurtransferase) is expressed at high levels in liver and is involved in the detoxification of cyanide. The full-length cDNA corresponding to the mouse rhodanese gene (Tst), which is located on chromosome 15, was cloned by PCR amplification of a liver cDNA library and subjected to DNA sequencing. Alignment of the rhodanese cDNA sequences from mouse and rat, which we previously cloned (Biochem. J. 275:227-231), revealed 97.3 percent identity at the protein level and 94.6 percent identity at the DNA level. When the mouse and rat cDNAs were expressed under the control of IPTG-inducible promoters in E. coli, the cell extracts exhibited cyanide-metabolizing activity, indicating that both genes encode functional rhodanese molecules. PMID- 7488187 TI - Formation of malondialdehyde-modified 2'-deoxyguanosinyl adduct from metabolism of chloral hydrate by mouse liver microsomes. AB - We previously reported that metabolism of chloral hydrate (CH), a widely used sedative and hypnotic, by male B6C3F1 mouse liver microsomes resulted in lipid peroxidation, producing the tumorigen malondialdehyde (MDA). Now we have found that incubation of CH in the presence of calf thymus DNA resulted in the formation of an MDA-modified DNA adduct as detected by 32P-postlabeling analysis. Similar results were obtained from incubation of trichloroacetic acid and trichloroethanol, both metabolites of CH. PMID- 7488189 TI - Effect of microinjection of rap 2A point mutant proteins on maturation and mottling in Xenopus oocytes. AB - We have analyzed the morphological alterations induced in Xenopus oocytes as a consequence of the microinjection of rap 2A proteins with point mutations at critical positions in the functional domains defined for members of the ras superfamily. Ala 35 rap 2A, a point mutation in the "effector" domain, is able to induce the rearrangement of the pigments in the animal hemisphere termed "mottling". This characteristic phenotype is delayed in asn 17 rap 2A and wild type rap 2A proteins. A completely different phenotype appears as a consequence of the microinjection of Val 12-rap 2A. This mutant is able to trigger oocyte maturation and is less efficient inducing mottling than Ala 35 or wild-type rap 2A proteins. Neither of these rap 2A mutant proteins is able to counteract ras induced maturation. However there is a striking interference with progesterone induced maturation, since wild-type or val 12 rap 2A microinjection reduced the viability of progesterone-treated oocytes. These findings suggest that rap2A proteins are regulatory elements in distinct signal transduction pathways that either lead to the morphological changes associated with the phenotype "mottling" or to oocyte maturation. PMID- 7488188 TI - The signal modulator protein 14-3-3 is a target of sphingosine- or N,N dimethylsphingosine-dependent kinase in 3T3(A31) cells. AB - Phosphorylation of a 28 kDa protein in 3T3(A31) cells upon addition of semi purified kinase was induced only by sphingosine (Sph) or N,N-dimethylsphingosine (DMS), but not by 17 other lipids tested. The 28 kDa protein, a major target of Sph- or DMS-dependent kinase, is now identified as 14-3-3 protein, which modulates a number of key protein kinases involved in regulation of intracellular signaling (Aitken A, Trends Biochem Sci 20:95-97, 1995; Morrison D, Science 266:56-57, 1994). Thus, Sph and DMS modulate the function of a versatile modulator controlling a number of signaling molecules. PMID- 7488191 TI - Dbl expression driven by the neuron specific enolase promoter induces tumor formation in transgenic mice with a p53(+/-) genetic background. AB - The dbl oncogene, generated by the truncation of the amino-terminal portion of the proto-oncogene sequence, encodes a guanine-nucleotide-releasing factor. The transforming activity of this oncogene has never been demonstrated in vivo or in vitro except in the NIH 3T3 mouse fibroblast cell line. The expression of the proto-dbl transcript is confined to tissues and tumors of neuroectodermal derivation. Therefore, to study the transforming activity of the dbl oncogene in vivo, we have generated transgenic mice that express this oncogene in neuroepithelial tissues. Mice carrying the dbl oncogene did not develop a tumor. Successively, to establish whether dbl interacts with the tumor suppressor gene p53 in tumorigenesis, we have used a p53 deficient mouse strain. The results reported here indicate that dbl is capable of causing tumor formation in vivo when its expression is driven in an appropriate cellular and genetic environment. PMID- 7488190 TI - Altered cAMP levels in retinas from transgenic mice expressing a rhodopsin mutant. AB - Transgenic mice expressing the rhodopsin mutant Pro347Ser (Serine 6) display retinal degeneration through apoptosis that is characteristic of the disease retinitis pigmentosa. By 5 weeks after birth, these mice have lost approximately 35% of their photoreceptor cells. Retinas from these mice showed higher levels of cAMP compared to the levels in retinas of normal mice. Our studies provide evidence that elevated cAMP is common to the apoptotic process that occurs in retinitis pigmentosa. In addition, in vitro studies demonstrate no differences in the ability of the mutant and the wild-type rhodopsin to activate transducin, the rod cell G protein, to be phosphorylated by rhodopsin kinase or to bind arrestin. Mutants of rhodopsin, including Pro347Ser, are mistargeted to the rod inner segment, raising the possibility that rhodopsin triggers apoptosis through activation of signaling pathways not normally under its control. PMID- 7488192 TI - Structural organization and chromosomal localization of the human nuclear gene (NDUFV2) for the 24-kDa iron-sulfur subunit of complex I in mitochondrial respiratory chain. AB - The structural organization of the entire human nuclear encoded gene for the 24 kDa iron-sulfur subunit of mitochondrial NADH-ubiquinone oxidoreductase (Complex I) and its chromosomal localization were determined. The gene contains 8 exons spanning 31.5 kb. The 5' flanking region sequenced lacks typical CAAT and TATA boxes but contains three putative GC boxes and there is one GC box at the beginning of the first intron. The sequences matching completely with the NRF-1 binding site and Mt elements were not identified in the flanking region. This gene was assigned to human chromosome 18 at region p11.3, by fluorescent in situ hybridization. PMID- 7488193 TI - Increased activity of porcine pancreatic phospholipase A2 by designed long-range electrostatic stabilisation of the transition state. AB - Stabilisation of the catalytic transition state by long-range charge interactions has been tested with mutagenesis for porcine pancreatic phospholipase A2. Electrostatics calculations were used to determine locations which would interact preferentially with one part of the dipolar charge separation that is believed to develop in the transition state. Experiment shows increased enzyme activity relative to wild-type recombinant enzyme for mutants N97D and N101D, consistent with the design. PMID- 7488194 TI - Fibronectin binding domain of P. gingivalis fimbriae. AB - P. gingivalis fimbriae play an important role in attachment of bacteria to various salivary components as well as to host cells and matrix proteins including fibronectin. In the present study, we investigated the binding domain of P. gingivalis fimbriae to fibronectin using synthetic peptides. A series of 20 mer fimbrillin peptides were used. Binding of fibronectin to purified fimbriae and synthetic peptides was assayed using polyclonal fibronectin antibodies as well as iodinated fibronectin. Purified fimbriae and peptide 126-146 (RMAFTEIKVQMSAAYDNIYTF) showed high levels of binding to fibronectin, while peptide 318-337(HLNVQCTVAEWVLVGQNATW) showed low but statistically significant binding. Our results suggest V-Q-X-X-X-A or V-X-X-X A common domain/domains present in both peptides might be involved in protein-protein interaction between P. gingivalis fimbriae and fibronectin. PMID- 7488195 TI - Up-regulation of glutathione S-transferases alpha by interleukin 4 in human hepatocytes in primary culture. AB - During inflammation and infection, overexpression of cytokines is associated with changes in cytochrome P450 (CYP) activities. The present study investigated the effect of cytokines on expression of the glutathione S-transferases (GST), phase II enzymes, involved in drug detoxication and in protection against lipid peroxidation. Human hepatocytes in primary culture were exposed to interleukin 6 (IL6), a proinflammatory cytokine and interleukin 4 (IL4) thought to be an anti inflammatory cytokine and known to induce CYP2E1 specifically. After a three-day treatment, no reproducible effects of IL-6 could be demonstrated on either GSTA1 and/or A2 or M1 mRNA levels (GSTA1 and A2 were not discriminated by the cDNA probe). In contrast, GSTA1 and/or A2 mRNAs and GSTA1 and A2 proteins were reproducibly increased after IL4 treatment. This increase was blocked by alpha amanitin, suggesting that active transcription is necessary and was associated with increased AP1 binding activities. These results provide evidences that IL4 exerts important effects on detoxifying hepatic drug metabolizing enzymes. PMID- 7488196 TI - Detection of high density lipoprotein binding proteins with a new method using apolipoprotein AI-DNP as a ligand. AB - To detect high density lipoprotein binding proteins, a new ligand blotting method that uses apolipoprotein AI-dinitrophenol as a ligand was established. Apolipoprotein AI-dinitrophenol was prepared by incubating the mixture of apolipoprotein AI and 2,4-dinitrobenzene sulfonic acid sodium salt at pH 8.5. Four to six dinitrophenol groups were found to be conjugated with an apolipoprotein AI molecule without any obvious impairment of its binding activity to the binding proteins. By this detection procedure, we detected two apolipoprotein AI binding proteins in plasma membrane fractions from bovine liver and adrenal cortex with molecular masses of 120 kDa and 95 kDa. Since this new ligand blotting method is simple and time-saving, it is expected to become a useful tool in the studies on high density lipoprotein binding proteins. PMID- 7488197 TI - HPLC analysis of 5-FU and FdUMP in tissue and serum. AB - Tissue and serum concentrations of 5-fluorouracil (5-FU) and 5-fluoro-2' deoxyuridine 5' monophosphate (FdUMP) can be determined using a simple and sensitive reverse phase HPLC technique. Samples are extracted with ethyl acetate, evaporated to dryness, resuspended in the mobile phase (water/methanol [85:15]), and injected into a C18 muBondapack column (250 mm x 30 mm) with a flow rate of 1 ml/min. Both fluorescence (266 lambda Excitation and 350 lambda Emission) and UV (250 nm) are used for detection. Standard solutions of 5-FU and FdUMP produce a linear relationship (r2 = 0.985) with a detection sensitivity of 50 pg. Recovery following this extraction procedure is 78%. This analytical method is simple and convenient to use for the simultaneous detection of 5-FU and FdUMP. PMID- 7488198 TI - Affinity of Bandeiraea (Griffonia) simplicifolia lectin-I, isolectin B4 for Gal alpha 1-->4 Gal ligand. AB - The affinity of Bandeiraea (Griffonia) simplicifolia lectin-I isolectin B4 (BSI B4) for the isomer of human blood group B active disaccharide (B, Gal alpha 1- >3Gal), the Gal alpha 1-->4Gal galabiose ligand, was studied by quantitative precipitin (QPA) and precipitin-inhibition assays. When human blood group B, P1 and H active glycoproteins were tested by OPA. BSI-B4 reacted strongly with both the B active glycoprotein purified from human ovarian cyst fluid and a P1 active glycoprotein isolated from sheep hydatid fluid and precipitated over 86% of the lectin nitrogen added. The P1 active glycoprotein-BSI-B4 interaction was inhibited by both Gal alpha 1-->3Gal alpha 1-->methyl and Gal alpha 1-->4Gal disaccharide indicating that BSI-B4 is not only reacting with Gal alpha 1-->3Gal disaccharide, but also recognizing Gal alpha 1-->4Gal. The galabiose sequence is frequently found in the carbohydrate chains of many glycosphingolipids located at the mammalian cell membranes such as intestinal and red blood cell membranes, for E. coli ligand binding and toxin attachment. PMID- 7488199 TI - A synthetic sialic acid analog that is resistant to the receptor-destroying enzyme can be used by influenza C virus as a receptor determinant for infection of cells. AB - A synthetic sialic acid analog, N-acetyl-9-acetamido-9-deoxy-neuraminic acid, can be used by influenza C virus as a receptor determinant for attachment to cells. In contrast to the natural determinant, N-acetyl-9-O-acetylneuraminic acid, the synthetic sialic acid is resistant to the action of the receptor-destroying acetylesterase of this virus. The sialic acid analog was enzymatically transferred to the surface of Madin-Darby canine kidney cells that are resistant to infection because of a lack of receptors. Influenza C virus was able to infect the modified cells through requiring a 10-fold larger amount of the sialic acid analogue on the cell surface compared to the natural receptor determinant. The quantitative difference is accounted for mainly by a less efficient binding of influenza C virus to the analog. Thus, in our system, inactivation of the receptor by the viral esterase is not required for the initiation of an influenza C virus infection. PMID- 7488200 TI - Physical association of Ras and cathepsins B and L in the conditioned medium of v Ha-ras-transformed NIH3T3 cells. AB - Immunoblot analysis of the conditioned medium of v-Ha-ras-transformed NIH3T3 cells showed that Ras protein is secreted in the culture fluid. Because cathepsins L and B whose activities are inhibited by Ras are also secreted, possible association between Ras and cathepsins was investigated by immunoprecipitation and following immunoblot analysis. The results showed that Ras is physically associated with cathepsins B and L in the culture fluid. PMID- 7488201 TI - A novel mitochondrial DNA point mutation associated with mitochondrial encephalocardiomyopathy. AB - A novel mitochondrial DNA (mtDNA) mutation at position nt 4320 in the tRNA(Ile) gene was associated with severe encephalopathy in a 7-month-old infant, who died of intractable hypertrophic cardiomyopathy. The mutation was present in heteroplasmic fashion (88%) in muscle and fulfills accepted criteria for pathogenicity. This is the fourth pathogenic mutation identified in this gene, which appears to be a "hotspot" for deleterious mutations affecting the heart. This report adds to the evidence of genetic heterogeneity in hypertrophic cardiomyopathies. PMID- 7488203 TI - Dolichyl phosphate, a potent inducer of apoptosis in rat glioma C6 cells. AB - Exposure of rat glioma C6 cells to dolichyl phosphate resulted in cell shrinkage followed by nuclear fragmentation and internucleosomal cleavage of genomic DNA, yielding ladder patterns of oligonucleosomal fragments, all characteristics of apoptosis. This phenomenon occurred in a dose and time dependent manner. Dolichol and prenol failed to induce apoptosis. The inhibitors of N-glycosylation, tunicamycin and swainsonine had no apparent effect on dolichyl phosphate-induced apoptosis. Apoptotic changes were also observed in HL-60 cells, SIRC cells and HeLa cells. Thus, dolichyl phosphate functions as a potential apoptosis inducer as well as an essential carrier lipid in the biosynthesis of N-linked glycoprotein. PMID- 7488202 TI - Metal uptake of recombinant cambialistic superoxide dismutase from Propionibacterium shermanii is affected by growth conditions of host Escherichia coli cells. AB - We constructed the complete nucleotide sequence coding for the cambialistic superoxide dismutase from Propionibacterium shermanii by ligation of a synthetic linker to a polymerase chain reaction amplification product obtained using degenerate primers. We set up an expression system yielding large amounts of recombinant superoxide dismutase in the cytoplasm of Escherichia coli and purified the enzyme from cells grown in a complex medium. The physicochemical properties of the recombinant enzyme were identical to those of the natural protein. Under anaerobic conditions the enzyme produced in an iron-supplemented medium incorporated iron as metal cofactor, while the enzyme purified from cells grown under aerobic conditions contained a variable amount of iron and manganese depending on metal availability. Functional equivalence of the two metals in this superoxide dismutase variant was indicated by independence of enzyme activity from Fe/Mn ratio. PMID- 7488204 TI - Copper-induced lipid oxidation in unfractionated plasma: the lag preceding oxidation as a measure of oxidation-resistance. AB - In spite of the pathological significance of oxidation of plasma lipids, no method is currently available for the evaluation of the susceptibility of these lipids to oxidation in unfractionated plasma. Here we demonstrate that copper induced oxidation of diluted plasma, in the presence of citrate can be monitored continuously by recording the absorbance at 245 nm. The kinetics of accumulation of oxidation products in unfractionated plasma is a sum of lipid oxidation products obtained in low and high density lipoproteins isolated from the same plasma. The kinetic profiles are reproducible and can be performed with plasma samples even after prolonged storage at 4 degrees C (up to two months) or after freezing and thawing of the plasma. Being simple and reproducible, yet correlating with the oxidizability of low and high density lipoproteins, this method can be used to evaluate the "oxidation resistance" of plasma lipids and thus serve as a standard index of the susceptibility of the plasma lipids of patients to oxidation-inflicted pathologies, including atherosclerosis. PMID- 7488206 TI - Different effects on the expression of CYP7 and CYP27 in rabbit liver by cholic acid and cholestyramine. AB - It was recently reported that both cholesterol 7 alpha-hydroxylase CYP7 and sterol 27-hydroxylase CYP27 are subject to negative feedback control by bile acids in isolated rat hepatocytes and rat liver (Twisk et al., (1995) Biochem. J. 305, 505-511; Vlahcevic et al. XIII International Bile Acid Meeting, September 30 October 2, 1994, Abstract 17). In the present work a series of experiments was performed to study whether there is a coordinate regulation of CYP7 and CYP27 also in rabbit liver. Treatments of rabbits with cholic acid or cholestyramine resulted in marked suppression and induction, respectively, of CYP7 mRNA. In contrast, there were no significant effects on the CYP27 mRNA expression, amount of CYP27 protein or mitochondrial 27-hydroxylase activity. Thus, there is an apparent difference between rat and rabbit with respect to regulation of CYP27 by bile acids. It is concluded that, in rabbit, there is little or no coordinate regulation of CYP7 and CYP27 at a transcriptional level, and that CYP27 is not subject to a negative feedback control by bile acids neither at a transcriptional nor at a posttranscriptional level. PMID- 7488207 TI - Biochemical characterization and posttranslational modification of AlgU, a regulator of stress response in Pseudomonas aeruginosa. AB - AlgU is homologous to the extreme heat shock sigma factor sigma E from enteric bacteria. In this work, AlgU was overproduced and purified and its function investigated at the biochemical level. AlgU was shown to associate with RNA polymerase and direct transcription of a target promoter. AlgU also exhibited multiple isoforms detected by 2D gel analysis. Treatment with a Ser/Thr phosphatase shifted the distribution of isoforms towards the basic side on 2D gels, suggesting that posttranslational modifications of AlgU may involve phosphorylation. The underphosphorylated forms of AlgU copurified with RNA polymerase. It is possible that phosphorylation affects AlgU activity or its stability. PMID- 7488205 TI - Overexpression of prostacyclin synthase inhibits growth of vascular smooth muscle cells. AB - To define the local effects of prostacyclin (PGI2) on the growth of vascular smooth muscle cells (VSMC), we transfected VSMC with an expression vector harboring the cDNA for PGI2 synthase (PGIS), which catalyzes the rearrangement of prostaglandin H2 to PGI2. Transfection of the human PGIS cDNA into rat VSMC did not affect DNA synthesis under serum-free basal conditions, but it increased PGI2 synthesis and decreased DNA synthesis under serum-stimulated conditions (in the presence of 1 or 5% fetal calf serum). These results demonstrated that locally synthesized PGI2 can exert autocrine and/or paracrine inhibitory effects on VSMC growth. It was also suggested that in vivo transfer of PGIS gene may be useful for the gene therapy for vascular disease such as neointimal hyperplasia. PMID- 7488209 TI - Reemergence of the fetal pattern of L-type calcium channel gene expression in non infarcted myocardium during left ventricular remodeling. AB - The cardiac L-type voltage-dependent calcium channel (VDCC) is a critical component of cardiac action potential and excitation-contraction coupling. The objective of the present study was to examine the changes in expression in Motif IV, an alternatively spliced region of the alpha-1 subunit of the VDCC channel in postmyocardial infarction (MI) remodeled rat left ventricle. RNase protection assay was used to determine alteration in isoform expression in the noninfarcted hypertrophied ventricular myocardium 21 days post myocardial infarction. Our study demonstrates that cardiac hypertrophy is associated with significant increase in the mRNA level of the fetal isoform, with the reversion of fetal:adult isoform ratio to the fetal phenotype. Changes in isoform expression in the post-MI remodeled ventricle, not previously reported, is a pertinent genetic marker of cardiac hypertrophy. PMID- 7488208 TI - Sequence analysis of frog alpha B-crystallin cDNA: sequence homology and evolutionary comparison of alpha A, alpha B and heat shock proteins. AB - alpha-Crystallin is a major lens protein present in the lenses of all vertebrate species. Recent studies have revealed that bovine alpha-crystallins possess genuine chaperone activity similar to small heat-shock proteins. In order to facilitate the determination of the primary sequence of amphibian alpha B crystallin, cDNA encoding alpha B subunit chain was amplified using a new "Rapid Amplification of cDNA Ends" (RACE) protocol of Polymerase Chain Reaction (PCR). PCR-amplified product corresponding to alpha B subunit was then subcloned into pUC18 vector and transformed into E. coli strain JM109. Plasmids purified from the positive clones were prepared for nucleotide sequencing by the automatic fluorescence-based dideoxynucleotide chain-termination method. Sequencing more than five clones containing DNA inserts coding for alpha B-crystallin subunit constructed only one complete full-length reading frame of 522 base pairs similar to that of alpha A subunit, covering a deduced protein sequence of 173 amino acids including the universal translation-initiating methionine. The frog alpha B crystallin shows 69, 66 and 56% whereas alpha A crystallin shows 83, 81 and 69% sequence similarity to the homologous chains of bovine, chicken and dogfish, respectively, revealing a more divergent structural relationship among these alpha B subunits as compared to alpha A subunits. Structural analysis and comparison of alpha A- and alpha B-crystallin subunits from eye lenses of different classes of vertebrates also shed some light on the evolutionary relatedness between alpha B/alpha A crystallins and the small heat-shock proteins. PMID- 7488211 TI - Identification of a novel 85-kDa lipoprotein lipase binding protein on human aortic endothelial cell surface. AB - Lipoprotein lipase (LPL), bound to the luminal surface of vascular endothelium catalyzes lipoprotein triglyceride hydrolysis. Studies were performed to identify human aortic endothelial (HAEC) cell-surface proteins having high affinity for LPL. LPL-sepharose affinity chromatography of [35S]O4 labeled HAEC proteins identified a 220-kDa proteoglycan. Ligand blotting of HAEC plasma membrane proteins with LPL revealed two specific binding proteins of MW 116 kDa and 85 kDa, respectively, which were not released from the cell-surface by heparin treatment. Since the 220-kDa and 116-kDa proteins have been reported previously in bovine endothelial cells, we focused on the 85-kDa protein. The 85-kDa protein was not labelled by incubation of the cells with [35S]O4, suggesting that it is not a sulfated proteoglycan. Treatment of HAEC with tunicamycin markedly decreased detection of the 85-kDa protein, suggesting that it is likely a glycoprotein synthesized by HAEC. We conclude that HAEC cell surface has three specific LPL binding proteins, a 220-kDa proteoglycan, a 116-kDa protein and a novel 85-kDa protein. PMID- 7488210 TI - Isozyme analysis of human normal polymorphonuclear leukocyte phosphofructokinase. AB - Phosphofructokinase (PFK) from human polymorphonuclear leukocytes (PMN) was characterized by immunological titration with subunit specific antibodies, column chromatography on QAE-Sephadex and SDS-polyacrylamide gel electrophoresis. Two different isozymes, M-type and L-type, were found. The M(r) values of the M and L subunits were 79,500 +/- 1,914 and 74,250 +/- 1,258, respectively. The two isozymes presented different kinetic and regulatory properties. The results suggest that PFK from human normal PMN is a mixture of M-type and L-type homotetramers, mainly, with possible minor heterotetrameric forms. PMID- 7488212 TI - Identification of a critical aspartate residue in transmembrane domain three necessary for the binding of somatostatin to the somatostatin receptor SSTR2. AB - To determine which residues within the rat somatostatin receptor subtype SSTR2 may be interacting with the lys9 of somatostatin-14 (S-14), mutant SSTR2 receptors were created by mutating asp89 or asp122. [125I Tyr11]S-14 binding to D89A and D89E mutants suggests that asp89 is not directly involved in S-14 binding. Binding studies with the charge switch mutants, asp9S-14, and D122K, suggest that asp122 may be interacting with the lys9 of S-14. [125I Tyr11]asp9S 14 displayed saturable binding to D122K with an affinity comparable to that seen with [125I Tyr11]S-14 and WT SSTR2. These data suggest that the interaction between lys9 of S-14 and the TM3 asp122 of SSTR2 represents one contact site between S-14 and SSTR2. PMID- 7488213 TI - Mu opioid receptor gene expression in immune cells. AB - We have previously demonstrated that administering morphine sulfate to rhesus monkeys alters the cell-mediated as well as humoral immune responses of these primates. Furthermore, morphine treatment greatly reduces the chemotactic and phagocytotic activities of primate polymorphonuclear (PMN) cells. The present study describes the identification and isolation of mRNA encoding the mu opioid receptor gene sequence from human and monkey immune cells. Through the use of primer sequences designed from the human brain mu opioid receptor cDNA sequence, specific opioid receptor segments in mRNA transcripts were amplified, cloned, and sequenced. The mu opioid receptor gene was therefore found expressed in the following cell types: CEM x174 (a hybrid of human T and B cells), Raji (human B cells), human CD4+ cells, human monocytes/macrophages, human PMN, monkey peripheral blood mononuclear cells (PBMC), and monkey PMN. These studies present the first evidence to demonstrate that cells of human and monkey immune systems constitutively express mu opioid receptor mRNA. PMID- 7488214 TI - Small pH gradients inhibit cytochrome c oxidase: implications for H+ entry to the binuclear center. AB - Respiring cytochrome c oxidase proteoliposomes generate internal alkalinity (delta pH) and a membrane potential (delta psi). Valinomycin collapses delta psi, increases delta pH, and slows steady state respiration. If delta pH is heterogeneously expressed trapped probes will underestimate it. Internal pH changes were therefore followed in COV containing two buffer systems of differing pKs. The alkalinization rate at pH 7 was unaffected by adding AMPSO (pK 9.0) to the usual internal HEPES (pK 7.5). At higher pH, AMPSO slowed the approach to steady state. delta pH inhibition is therefore not due to a large alkalinization in a small COV fraction. The O2-reducing center may move protons via a local aqueous phase that is near electrical and pH equilibrium with the phase inside the COV. The dielectric in this membrane region can put the center electrically 'inside' even though it is physically 'outside'. PMID- 7488215 TI - Identification of the major lipoproteins in crayfish hemolymph as proteins involved in immune recognition and clotting. AB - Lipid-containing hemolymph proteins from males of the crayfish Pacifastacus leniusculus were isolated by density gradient ultracentrifugation. Two major lipoproteins, one high density lipoprotein (HDL) and one very high density lipoprotein (VHDL), were characterized. The HDL and the VHDL were found to be identical to two proteins previously studied for their roles in immune recognition and hemolymph clotting, namely the beta-1,3-glucan binding protein and the clotting protein. These results imply that crayfish lipoproteins have dual functions, and that they are involved in immunity, hemolymph clotting, and lipid transport in these animals. Also, the oxygen-transporting protein hemocyanin was found to have a small lipid content. PMID- 7488216 TI - The liver-specific response to phenobarbital involves a transient increase in phosphorylation of a 34-kda nuclear protein in rat liver and in hepatocytes in culture. AB - We have examined the influence of phenobarbital, a liver detoxication gene inducer and a potent tumor promoter, on the phosphorylation status of nuclear enriched proteins in primary rat hepatocyte cultures and in whole livers. Freshly isolated cells were plated on plastic dishes in presence of serum for 4 h and 2 mM phenobarbital was added for various times, following serum withdrawal. A transient increase in phosphorylation of a 34-kda nuclear protein was detected at 6 h. In whole livers, but not in kidneys, a nuclear protein with the same electrophoretic mobility was also transiently over-phosphorylated, following injection of 80 mg/kg phenobarbital, although the peak activity was attained after 30 min only. No immunological relatedness between major histones and the 34 kda protein was found. Our results demonstrate a specific, yet undescribed, transient effect of phenobarbital on the phosphorylation status of a 34-kda nuclei-enriched protein in rat hepatocytes and in rat liver. PMID- 7488217 TI - cDNA cloning and the identification of an aggrecanase-like cleavage site in rat brevican. AB - Brevican is a member of the aggrecan family of chondroitin sulfate proteoglycans that is predominantly expressed in the nervous system. In the adult brain, the brevican core protein undergoes proteolytic cleavage and exists as a 145-kDa full length form and an 80-kDa C-terminal fragment. We have determined the complete primary structure of rat brevican and the N-terminal amino acid sequence of the 80-kDa fragment. The results demonstrate that the cleavage of the brevican core protein occurs in a highly conserved region within a generally poorly conserved central domain, and that the sequence at the cleavage site shows intriguing similarities to the "aggrecanase" cleavage site in aggrecan. cDNA cloning of rat brevican has also revealed that the putative hyaluronic acid binding protein, BEHAB, is identical to the N-terminal half of brevican. PMID- 7488218 TI - Mitogenic response to TGF-beta in 3T3-F442A cells. AB - In 3T3-F442A cells, TGF-beta caused cellular proliferation in a time and dose dependent manner. TGF-beta induced cyclin D1 and cdk2 proteins in 3T3-F442A cells. The mitogenic effect of TGF-beta was specific in nature. The antimitogenic agent, hGH, inhibited the mitogenic effect of TGF-beta and was associated with inhibition of cyclin D1 expression. The protein kinase c inhibitor, staurosporine, inhibited the mitogenic effect of TGF-beta. Taken together, these results suggest that TGF-beta affects expression levels of cell cycle-regulated proteins and its mitogenic effect is mediated through protein kinase C in 3T3 F442A cells. PMID- 7488219 TI - Inhibition of the thiol-specific antioxidant activity of rat liver MSP23 protein by hemin. AB - Mouse stress-inducible 23-kDa protein (MSP23) belongs to a new type of antioxidative protein family and is highly expressed in livers. To examine its physiological role in the tissue, we have purified native MSP23 protein from the cytosolic fraction of rat liver. The purified protein showed the thiol-specific antioxidant activity and protected glutamine synthetase from inactivation by a mixed metal-thiol oxidation. We examined the effect of hemin on that activity since it was recently shown by others that the rat liver MSP23 protein had a high binding affinity to heme. We have found that hemin at low concentrations inhibits the antioxidative activity of MSP23. This result suggests that one type of oxidant damage caused by hemin to cells is due to inactivation of the antioxidative protein, MSP23. PMID- 7488220 TI - Why is phosphonodifluoromethyl phenylalanine a more potent inhibitory moiety than phosphonomethyl phenylalanine toward protein-tyrosine phosphatases? AB - The phosphonodifluoromethyl phenylalanine (F2Pmp) is superior to phosphonomethyl phenylalanine (Pmp) as a non-hydrolyzable phosphotyrosine (pTyr) mimetic. The difluoromethyl moiety increases the inhibitory potency of a F2Pmp-containing peptide over a Pmp-containing counterpart by 1000-fold toward the protein tyrosine phosphatase (PTPase), PTP1. Fluorine substitution at the methylene carbon have the double effect of lowering the phosphonate pKa2 as well as introducing hydrogen bonding interactions similar to the phosphate ester oxygen in pTyr. The inhibition of PTP1-catalyzed dephosphorylation reaction by both the F2Pmp and Pmp-containing peptides did not vary as a function of pH. The data indicate that both the monoanion and the dianion forms of the phosphonate bind PTP1 with equal efficiency. Thus, the better binding by the F2Pmp-peptide as compared to the Pmp-peptide is not due to the difference in pKa2. Taken together, these results offer an explanation for the increased affinity of F2Pmp for PTP1. The two fluorine atoms in F2Pmp may be able to interact with active site residues in PTP1 in a fashion analogous to that involving the phenolic oxygen and side chains in the active site of PTP1. Ki measurements for a simple phosphonic acid, Pmp- and F2Pmp-containing peptides suggest that although the principal recognition element is F2Pmp itself, the surrounding amino acids are required for high affinity binding. Comparative analysis of the inhibition of PTP1, PTP alpha and LAR by F2Pmp-containing peptides suggests that selective, tight-binding PTPase inhibitors can be developed. PMID- 7488221 TI - Cloning of the gene encoding a putative serine/threonine protein kinase which enhances spermine uptake in Saccharomyces cerevisiae. AB - Polyamine uptake in Saccharomyces cerevisiae was modulated by extracellular magnesium; in a magnesium-limited medium, polyamine, especially spermine, was overaccumulated into the interior, whose level was then toxic for the growth of this organism (Maruyama, T., Masuda, N., Kakinuma, Y., and Igarashi K. (1994) Biochim. Biophys. Acta 1194, 289-295). Here we isolated a mutant (strain YTM22-8) whose growth was tolerant to spermine in magnesium-limited medium. This mutant was defective in polyamine uptake and did not overaccumulate spermine. From a yeast genome library we cloned a gene (POT1) which restored the spermine uptake of this mutant and the sensitivity of the growth to spermine. The nucleotide sequence of the POT1 gene indicated that it encodes a putative serine/threonine protein kinase and is located on chromosome XI. The results suggest that spermine uptake by this organism is probably regulated by phosphorylation and dephosphorylation. PMID- 7488222 TI - Glucose-6-phosphate dehydrogenase from Saccharomyces cerevisiae is a glycoprotein. AB - A commercial preparation of glucose-6-phosphate dehydrogenase (G6PD) purified from Saccharomyces cerevisiae was subjected to PAGE analysis under both nondenaturing and denaturing conditions. The enzyme, identified by both activity staining and anti-yeast G6PD antibody immunoblotting, was shown to contain carbohydrate using the highly specific periodate-digoxigenin antidigoxigenin method which is diagnostic for glycoproteins. PMID- 7488223 TI - Activation of signal transducer and activator of transcription-3 (Stat3) expression by interferon-gamma and interleukin-6 in hepatoma cells. AB - Signal Transducer and Activator of Transcription 3 (Stat3) is a latent protein activated in response to various cytokines and growth factors. It is believed that Stat3 is a key signaling molecule involved in the regulation of acute phase gene expression by interleukin 6 (IL-6) in hepatocytes. We report that both IL-6 and interferon gamma (IFN gamma) up-regulate the expression of Stat3 on both mRNA and protein levels in rat and human hepatoma cells. The effect of IL-6 and IFN gamma on Stat3 mRNA expression was time- and dose-dependent. Other factors, including IL-1, TNF alpha, EGF, Dexamethasone and PMA, did not have any effect on Stat3 mRNA expression. Moreover, we show that the rapid induction of Stat3 expression by IL-6 and IFN gamma was independent of ongoing protein synthesis, suggesting regulation by Stat3 and Stat1, respectively. PMID- 7488224 TI - Target-mediated resistance to beta-lactam antibiotics. PMID- 7488225 TI - Piracetam-induced changes to membrane physical properties. A combined approach by 31P nuclear magnetic resonance and conformational analysis. AB - Piracetam, Nootropil (2-oxo-1-pyrrolidine acetamide), is a drug promoting erythrocyte deformability. To establish the mode of action of this compound, we have investigated its influence on the organization of model phospholipid membranes. 31P NMR data show that the drug induces a structural modification in liposomes made of phosphatidylcholine and phosphatidylethanolamine. Our conformational analysis results have allowed the interpretation of the effect of piracetam on these model membranes: the specific interaction between the drug molecules and the phosphate headgroups induces a new organization of the lipids favouring formation of mobile drug-phospholipid complexes that exhibit an isotropic-type signal in the 31P NMR spectra. PMID- 7488226 TI - Multidrug resistance bypass in cells exposed to doxorubicin-loaded nanospheres. Absence of endocytosis. AB - Multidrug resistance bypass has been achieved in vitro with polyalkylcyanoacrylate nanospheres loaded with doxorubicin as previously shown by Cuvier et al. Fluorescence-imaging experiments were performed to determine if the endocytosis of the particles by the cells could be responsible for this activity. The results obtained with three lines of sensitive and resistant cells (K562, MCF7, and C6) demonstrate that the particles were not internalized by the cells, nor were they adsorbed onto the cell surface. In contrast, these nanospheres were very efficiently endocytosed by phagocytic cells such as macrophages. No correlation was observed between the fate of the particles, endocytosed or not, and their cytotoxic activity. It was concluded that no endocytosis step was involved in the mechanism of the multidrug resistance bypass obtained by associating a drug to polymeric particles. PMID- 7488227 TI - Increased killing of prostate, breast, colon, and lung tumor cells by the combination of inactivators of O6-alkylguanine-DNA alkyltransferase and N,N' bis(2-chloroethyl)-N-nitrosourea. AB - The ability of a number of compounds that act as inactivators of O6-alkylguanine DNA alkyltransferase (AGT) to sensitize human tumor cell lines to the effects of N,N'-bis(2-chloroethyl)-N-nitrosourea (BCNU) were examined. The AGT inactivators tested included O6-benzylguanine (BG) and its 8-aza-, 8-bromo-, 8-methyl-, 8-oxo, and 8-amino-derivatives and O6-[p-(hydroxymethyl)benzyl]guanine. All of these compounds except the 8-amino-derivative were active in greatly increasing the killing of HT29 colon, Du145 prostate, MCF-7 breast and A549 lung tumor cells by BCNU. Their activities were comparable to those of BG. Two pyrimidines, 2,4 diamino-6-benzyloxy-5-nitrosopyrimidine and 2,4-diamino-6-benzyloxy-5 nitropyrimidine, were found to be considerably more potent than BG in enhancing BCNU-induced cell killing. The addition of a steroid group to the 9-position of BG forming either O6-benzyl-9-[3-oxo-4-androsten-17 beta yloxycarbonyl)methyl]guanine or O6-benzyl-9-[3-oxo-5 alpha-androstan-17 beta yloxycarbonyl)methyl]guanine also produced compounds effective in enhancing the cytotoxicity of BCNU when added at 10 microM. These results indicate that a range of potent compounds with potentially different pharmacokinetics is available to test the hypothesis that inactivation of AGT overcomes the resistance of many tumor cells to nitrosoureas. PMID- 7488228 TI - Differential effect of 1-(5-isoquinolinesulfonyl)-2-methylpiperazine (H-7) on alpha B-crystallin and hsp70 gene expression in murine cell lines. AB - We studied the effect of isoquinolinesulfonamide derivatives (H-7, H-8, and HA1004) on the expression of two heat shock genes (alpha beta-crystallin and hsp70) in NIH 3T3 and Swiss 3T3 cells after heat shock at 45 degrees for 10 min. Western blots and northern blots showed that H-7 effectively suppressed the accumulation of HSP70 and alpha B-crystallin mRNA as well as the synthesis of their proteins. The degree of suppression was dependent upon the concentration of the drug. Moreover, the expression of the hsp genes was differentially suppressed by H-7. The expression of the alpha B-crystallin gene was more effectively inhibited than that of the hsp70 gene by H-7. Nuclear run-on assay demonstrates that this difference was due to the differential effect of H-7 on the elongation of transcription of different hsp genes. PMID- 7488229 TI - Biochemical pharmacology of penclomedine (NSC-338720). AB - Penclomedine (PEN) is a synthetic pyridine derivative that has been selected for clinical development based on its activity against human and mouse breast tumors implanted in mice. Its mechanism of action was unclear, and we were interested in determining its mechanism of cytotoxicity in vitro and in vivo. We found chromosome breaks, gaps, and exchanges in P388 ascites cells from BD2F1 mice treated with 200 mg/kg PEN. Maximal observed damage occurred 24 hr after drug administration. Alkaline elution indicated only limited DNA strand breaks and interstrand cross-linking. In vitro, PEN (75 micrograms/mL) inhibited RNA and DNA syntheses almost completely. In addition, incubation of [14C]PEN with rat liver S 9 fraction in the presence of calf thymus DNA resulted in the stable transfer of radioactivity to DNA. Addition of butylated hydroxytoluene, a free radical scavenger, to the incubation mixture inhibited the binding of drug to DNA, implicating free radicals as the ultimate reactive species. These data suggest that PEN can be metabolized to free radical, DNA-reactive products, and that its cytotoxicity is due to chromosomal damage produced by monofunctional alkylation. As an alternate mechanism, the ability of PEN to inhibit cellular dihydroorotate dehydrogenase was explored. Although PEN is an inhibitor of this enzyme in cells in vivo, in vitro, and in isolated cell sonicates, HPLC analyses of ribonucleotide triphosphate pools in P388 cells showed that all triphosphates had increased, especially UTP. Addition of uridine to the cell culture failed to prevent PEN-mediated cytotoxicity, suggesting that inhibition of de novo pyrimidine biosynthesis was not likely to be an important mechanism of action of this drug. These data suggest that PEN is activated in cells to a free radical that binds DNA. PMID- 7488230 TI - Evidence for the lack of hepatic N-acetyltransferase in suncus (Suncus murinus). AB - The abilities of liver cytosol fractions from the suncus and Sprague-Dawley (SD) rats to N-acetylate aniline, p-aminobenzoic acid, p-aminosalicylic acid and 2 aminofluorene (AF) were compared. The cytosol from rats N-acetylated these substrates at efficient rates, whereas the cytosol from the suncus did not N acetylate these substrates at detectable rates. When AF was given to the suncus, 2-acetylaminofluorene (AAF), a metabolite of AF formed by N-acetyltransferase (NAT), was not detectable in serum, whereas the metabolite was seen clearly in rats. Northern blot and Southern blot analyses, using cDNAs coding for human NATs as probes, indicated that not only the transcripts but also the genes of the enzymes were undetectable in suncus. These results suggest that the suncus is among the few species known to lack NATs. PMID- 7488231 TI - Modulation by benzo[a]pyrene of epidermal growth factor receptors, cell proliferation, and secretion of human chorionic gonadotropin in human placental cell lines. AB - Clinical observations indicate that maternal cigarette smoking has significant detrimental effects on fetoplacental development. The present study used human trophoblastic choriocarcinoma cell lines of placental origin to investigate the effects of benz[a]pyrene (BaP) on epidermal growth factor (EGF) receptors, cell proliferation and human chorionic gonadotropin (hCG) secretion. BaP decreased 125I-EGF binding and EGF receptor protein in a concentration-related manner in both BeWo and JEG-3 cell lines. The steady-state level of EGF receptor mRNA, however, was not changed significantly by BaP in either cell line. Cell proliferation was unchanged or slightly increased following exposure to 10 and 50 microM BaP in the presence of serum, whereas proliferation progressively decreased in cells exposed under serum-free conditions. The mitogenic effect of EGF was inhibited by cotreatment with BaP in both cell lines. Further study of trophoblast endocrine function showed that both basal and EGF-stimulated secretion of hCG was reduced significantly by BaP exposure in BeWo cells, whereas no adverse effect was seen in JEG-3 cells. Finally, cytochrome P450 1A1 (CYP1A1) was induced in a concentration-dependent manner by BaP in both cell lines. Thus, data indicate that the BaP-mediated loss of EGF receptors alters trophoblast proliferation and endocrine function, and that different mechanisms may be involved in the regulation of hCG secretion in BeWo and JEG-3 cells. In addition, this study supports the feasibility of using the BeWo and JEG-3 trophoblastic choriocarcinoma cell lines to investigate biomarkers and mechanisms of placental toxicity. PMID- 7488232 TI - Reduction of nitric oxide synthase activity in human neutrophils by oxidized low density lipoproteins. Reversal of the effect of oxidized low-density lipoproteins by high-density lipoproteins and L-arginine. AB - Oxidized low-density lipoproteins (ox-LDL) inhibit vascular relaxation by decreasing the synthesis or rapid degradation of endothelium-derived relaxing factor (EDRF), now identified to be nitric oxide (NO). We examined the regulation of NO synthase activity in human neutrophils, which also generate NO, by lipoproteins. Isolated human neutrophils were incubated with native-LDL, ox-LDL (10-50 micrograms protein/mL), high-density lipoproteins (HDL, 100 micrograms protein/mL) or HDL+ox-LDL, and NO synthase activity was measured as conversion of [3H]L-arginine to [3H]L-citrulline. Ox-LDL, but not native-LDL or HDL, significantly decreased NO synthase activity in human neutrophils. This effect of ox-LDL was incubation time and concentration dependent. The incubation of cells with HDL or L-arginine diminished the effects of ox-LDL on NO synthase activity. Thus, ox-LDL decreases the activity of NO synthase enzyme, and this effect of ox LDL can be modified by HDL and L-arginine. PMID- 7488234 TI - Regulation by 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD) of the DNA binding activity of transcriptional factors via nuclear protein phosphorylation in guinea pig adipose tissue. AB - 2,3,7,8-Tetrachloro-p-dioxin (TCDD) induced a modest stimulation of nuclear protein phosphorylation in explant tissue cultures in 10 min, followed by a substantial decrease in the level of total protein phosphorylation activity in the nucleus. Curiously, this TCDD-induced decline in nuclear protein phosphorylation was accompanied by an increase in cytosolic and extranuclear protein phosphorylation activity. One of the main causes for such a decrease in the protein phosphorylation activity in the nucleus appears to be related to some increase in protein phosphatase activities as judged by the counteractions of okadaic acid and Na3VO4 to the above effect. In addition, TCDD induced changes in nuclear protein kinase activities as well. Manganese-stimulated protein kinase was found to be the predominant type of nuclear protein phosphorylating activity affected by TCDD, with 60% of the total activity due to heparin-sensitive casein kinase II (CK II), a major nuclear protein kinase. The level of CK II activity in the nuclear protein preparation from adipose tissue of TCDD-treated guinea pigs (1 microgram/kg) in the presence of 100 nM heparin was only 35% of the control value after 24 hr. In addition, TCDD was found to increase the protein kinase C and microtubule-associated protein 2 kinase activities as early as 15 min after treatment in isolated adipose tissues in culture. Under in situ incubation conditions with explant tissues in culture, TCDD rapidly enhanced the DNA binding activity of the transcriptional factor AP-1, whereas the same treatment reduced c Myc DNA binding activity. Genistein, a specific protein tyrosine kinase inhibitor, abolished the stimulatory effect of TCDD on AP-1 binding activity, but not on DNA binding activity of c-Myc. Phorbol ester (TPA) increased the binding activity of AP-1 and c-Myc, as expected. However, TCDD in combination with TPA caused a slight reduction in binding activity of both transcriptional factors. On the other hand, in the presence of forskolin, the stimulatory effect of TCDD on AP-1 binding activity and the inhibitory effect on c-Myc were still apparent. Okadaic acid almost abolished the binding activity of c-Myc, whereas in combination with TCDD a stimulatory effect was found. These observations are consistent with the idea that TCDD regulates the DNA binding activity of AP-1 and c-Myc mainly through modulating their states of phosphorylation by altering protein kinase and phosphatase activities. PMID- 7488233 TI - Maitotoxin-elicited calcium influx in cultured cells. Effect of calcium-channel blockers. AB - Maitotoxin elicited a marked influx of 45Ca2+ into NIH 3T3 fibroblast cells. The influx was blocked by imidazoles (econazole, miconazole, SKF 96365, clotrimazole, calmidazolium) with IC50 values from 0.56 to 3 microM. Phenylalkylamines (verapamil, methoxyverapamil) and nitrendipine were less potent, and diltiazem was very weak. Among other calcium blockers, the diphenylbutylpiperidines fluspirilene and penfluridol, the diphenylpropylpiperidine loperamide, and the local anesthetic proadifen were quite active with IC50 values of 2-4 microM. The pattern of inhibition of maitotoxin-elicited calcium influx did not correspond to the ability of the agents to block elevation of calcium that ensues through calcium-release activated calcium (CRAC) channels after activation of phosphoinositide breakdown by ATP in HL-60 cells. The imidazoles did block CRAC channels, but fluspirilene, penfluridol, loperamide and proadifen were ineffective. Loperamide actually appeared to enhance influx of calcium via the activated CRAC channels. The imidazoles, in particular calmidazolium, caused an apparent influx of calcium and caused a stimulation of phosphoinositide breakdown in HL-60 cells. PMID- 7488235 TI - Competition between inhibitors of the trypanosome alternative oxidase (TAO) and reduced coenzyme Q9. AB - The trypanosome alternative oxidase (TAO) is an attractive target for chemotherapy for the diseases caused by African trypanosomes because there is no equivalent enzyme in mammalian hosts. Many inhibitors of this enzyme have been described, but there have been no data on the mechanism of inhibition. In the present study, reduced 2,3-dimethoxy-5-methyl-6-decyl-1,4-benzoquinone (decyl-CoQ H2) was used as a substitute for the natural substrate CoQ9-H2 to allow direct measurements of the TAO in crude mitochondrial preparations from Trypanosoma brucei brucei. A Km value of 3.8 microM was obtained for this substrate. The following five compounds that have alkyl side chains from 1 to 4 carbons and belong to three classes of inhibitors showed a competitive inhibition pattern with respect to decyl-CoQ-H: p-methoxybenzhydroxamic acid, p-ethoxybenzhydroxamic acid,p-n-butyloxybenzhydroxamic acid, methyl 3,4-dihydroxybenzoate and N-n-butyl 3,4-dihydroxybenzamide. The following four compounds belonging to the same chemical classes but having alkyl side chains from 10 to 12 carbons showed uncompetitive inhibition patterns: p-n-dodecyloxybenzhydroxamic acid, n-decyl 3,4 dihydroxybenzoate, n-dodecyl 3,4-dihydroxybenzoate, and N-n-decyl-3,4 dihydroxybenzamide. Clearly, the first group of inhibitors compete with CoQ-H2 for the active site of the TAO. We propose that the uncompetitive patterns produced by the second group of inhibitors are due to the greater lipophilicity of these compounds and the resulting change in the interaction of the inhibitors and the membrane containing the TAO, thus affecting the local concentration of the inhibitors at the active site. PMID- 7488236 TI - Synthesis and characterization of an aryl-azidoparoxetine. A novel photo-affinity probe for serotonin-transporter. AB - Paroxetine is an effective antidepressant drug and potent serotonin (5-HT) uptake inhibitor. It selectively labels 5-HT transporter on platelets and neurons. We report here the synthesis of an aryl-azido derivative of paroxetine, which is a novel photoactive and irreversible ligand for the [3H]paroxetine binding site on the platelet 5-HT transporter. The compound inhibited [3H]paroxetine binding (IC50, 55 nM) and 5-HT uptake (IC50, 12 nM) at equilibrium conditions and inactivated 10-20% of [3H]paroxetine binding sites upon irradiation at 320 nm. SDS-PAGE of platelet protein extract labelled with the radioactive analogue of the synthesized probe revealed the presence of four radioactive bands of which the 71-kDa one was the most prominent. PMID- 7488237 TI - Preferential requirement for protein tyrosine phosphatase activity in the 12-O tetradecanoylphorbol-13-acetate-induced differentiation of human colon cancer cells. AB - Some lines of colon cancer cells are forced to undergo differentiation by 12-O tetradecanoylphorbol-13-acetate (TPA). The increases in activities of both protein tyrosine phosphatase (PTP) and protein tyrosine kinase (PTK) have been reported to be associated with the TPA-induced differentiation of HL-60 leukemia cells. In the present study, a 2-fold increase in PTP activity was observed in SW620 human colon cancer cells after 30 min of TPA treatment; a maximal level (4- to 5-fold) was reached at 60 min and continued for more than 6 hr. In addition, two TPA-induced differentiated characteristics, morphological alteration and release of cellular surface proteoglycan, were effectively blocked by PTP inhibitors, such as sodium orthovanadate (50 microM), zinc chloride (100 microM), and iodoacetate (250 microM), but not by the protein serine/threonine phosphatase inhibitor okadaic acid (20 nM). On the other hand, although TPA induced a transient slight increase in PTK activity (1.4-fold) at 60 min, four PTK inhibitors (genistein, herbimycin A, tyrphostin-23 and quercetin) had different effects on the TPA-induced release of cell surface proteoglycan. Genistein (60 microM) potentiated this process, but in contrast, quercetin (45 microM) could partially inhibit the TPA effect. Taken together, these observations suggest that both PTP and PTK activities were increased in SW620 cells in response to TPA; however, the activation of PTP seems to be preferentially required for the TPA induced differentiation of SW620 human colon cancer cells. PMID- 7488238 TI - Enhanced biliary excretion of canalicular membrane enzymes in ethynylestradiol induced cholestasis. Effects of ursodeoxycholic acid administration. AB - Cholestasis is associated with a marked increase in the release of canalicular membrane enzymes into bile. This phenomenon has been related to an increased lability of these canalicular membrane integral proteins to the solubilizing effects of secreted bile salts. To further characterize the effects of oral ursodeoxycholic acid (UDCA) administration on ethynylestradiol (EE)-induced cholestasis, the influence of this bile acid on changes in biliary excretion of membrane-bound enzymes was investigated. Bile flow, basal bile salt and biliary lipid secretory rates, the maximum secretory rate of taurocholate (TC SRm), and the biliary excretion of the canalicular membrane-bound ectoenzymes alkaline phosphatase (ALP) and gamma-glutamyl transpeptidase (GGT) were measured in rats after EE and/or UDCA administration. The activities of ALP, GGT and Na+,K(+) ATPase in purified isolated canalicular and sinusoidal membrane fractions and the ultrastructure of hepatic acinus, including histochemical studies of ALP distribution, were also examined. EE significantly reduced bile flow, bile salt and biliary lipid secretory rates, and TC SRm, and caused dilatation and loss of microvilli at the canalicular pole of hepatocytes. Biliary excretion of ALP increased 2-fold, whereas biliary excretion of GGT was unchanged. The relationship between biliary excretion of ALP or GGT and bile salt secretion (units of enzyme activity secreted per nanomole of bile salt) was greater in EE treated rats compared with controls (2.1- and 1.5-fold greater for ALP and GGT, respectively), indicating that in EE-induced cholestasis more enzyme was released into bile per nanomole of bile salt. Na+,K(+)-ATPase activity in sinusoidal membrane fraction was reduced significantly, whereas ALP activity increased in both membrane fractions in EE-treated rats. The histochemical distribution of ALP in the acinus showed a strong reaction in acinar zone 3 and at both the canalicular and sinusoidal membranes. Oral administration of UDCA prevented EE induced bile secretory failure by normalizing bile flow, bile salt and biliary phospholipid secretory rates, and TC SRm. UDCA also prevented the EE-induced changes in the biliary excretion of enzymes. On the contrary, UDCA did not modify either the enzyme activity in isolated membrane fractions or the morphological or ALP histochemical changes associated with EE administration. These data indicate that in EE-induced cholestasis changes occur at the canalicular membrane, enabling this portion of the plasma membrane to be more susceptible to the solubilizing effect of bile salt, and that oral administration of UDCA prevents bile secretory failure and changes in the biliary excretion of ALP and GGT in EE treated rats. PMID- 7488239 TI - Influence of ethacrynic acid on glutathione S-transferase pi transcript and protein half-lives in human colon cancer cells. AB - Ethacrynic acid (EA) is a plant phenolic acid that is both an inhibitor and an inducer of glutathione S-transferase (GST) activity. To determine contributory factors in the increased GST activity caused by EA treatment, human colon carcinoma HT29 cells were compared with a cloned EA-resistant population (HT6-8) maintained in medium containing 72 microM EA. Several factors are involved in the increased expression of GST pi in HT6-8. For example, nuclear run-on experiments showed an approximately 2-fold increase in the rate of transcription of GST pi. In addition, the half-life of GST pi transcript was increased from 4.1 (wild type, HT29, HT4-1) to 8.4 hr. The half-life of GST pi protein was 1-2 hr in HT4-1 cells versus 8-9 hr in HT6-8 cells. When either human ovarian carcinoma cells (SKOV3) or human prostatic carcinoma cells (DU145) were treated with EA, the half life of the GST pi transcript was also increased. The transcript half-lives of another thiol-metabolism enzyme, gamma-glutamylcysteine synthetase (gamma-GCS), and a phase II detoxification enzyme, dihydrodiol dehydrogenase (DDH), were also increased in HT6-8, SKOV3 and DU145 cells treated with EA. However, the half lives of transcripts from "housekeeping genes," such as glyceraldehyde 3 phosphate dehydrogenase (G3PDH), beta-actin and beta-tubulin, were not changed in these cell lines following EA. Apparently, a number of coordinated factors are involved in EA-enhanced expression of GST pi and other detoxification enzymes. PMID- 7488240 TI - Cytotoxicity of BMS-181174. Effects of hypoxia, dicoumarol, and repair deficits. AB - The mitomycin C (MC) analog BMS-181174 (previously designated as BMY25067) has been shown to be active against a variety of solid tumors in mice. The activity of this compound against tumor cell lines resistant to MC and the different toxicity profiles of BMS-181174 and MC suggested that there may be significant differences in the metabolism and the mechanisms of action of these two compounds. Our studies with a mouse mammary tumor cell line (EMT6), a wild-type Chinese hamster cell line (AA8), and three repair-deficient Chinese hamster cell lines (UV4, UV5, and EM9) supported this concept. BMS-181174 was more toxic to all five cell lines in air than in hypoxia; in contrast, MC is more toxic in hypoxia. Dicoumarol (which increases the cytotoxicity of MC in hypoxia and reduces the cytotoxicity of this drug in air) did not alter the cytotoxicity of BMS-181174. This finding suggests that neither DT-diaphorase nor cytochrome b5 reductase is involved in the activation of BMS-181174. Studies with the repair deficient cell lines suggest that DNA strand breaks are not important to the cytotoxicity of BMS-181174, and that cross-links and adducts may be the critical lesions; these studies also suggest that the lethal lesions produced by BMS 181174 are the same under aerobic and hypoxic conditions. PMID- 7488241 TI - Structure-activity relationship of verapamil analogs and reversal of multidrug resistance. AB - We studied the relationship between the chemical structure and multidrug resistance (MDR) reversal activity of racemic verapamil (VER) and 14 VER analogs (VAs). The LoVo-R human colon carcinoma cell line was used as an experimental model. This cell line exhibited a typical MDR phenotype and overexpressed the MDR1 gene products. Key structural features were identified as being related to MDR reversal and cytotoxic activity. In particular, we demonstrated that the methoxy groups in the VER molecule structure [1.7-Bis-(3.4-dimethoxyphenyl)-3 methylaza-7-cyan-8-methyl-n onane] prevented cytotoxicity when the VAs were used alone, whereas the 7-cyan-8-methyl groups were important for MDR reversal activity and interaction with P-glycoprotein (P-gp). Among the VAs tested, the most active compounds were gallopamil, R-isomer of VER (R-VER), and nor-VER, which potentiated doxorubicin (DOX) cytotoxicity by 52.3 +/- 7.2 (n = 3 +/- SD), 38.9 +/- 6.4 (n = 4 +/- SD), and 35.4 +/- 4.3 (n = 3 +/- SD) times, respectively. The reversal activity of these compounds was similar to that of VER, which enhanced DOX cytotoxicity by 41.3 +/- 5.0 (n = 3 +/- SD) times. The potentiation of DOX cytotoxicity was associated with an increase in DOX uptake in LoVo-R cells and with an increased [3H]azidopine P-gp photolabeling inhibition. Some compounds that had a high reversal potency (i.e. R-VER and nor-VER) showed a lower calcium antagonist activity than VER, and seem useful candidates for the treatment of MDR in cancer patients. PMID- 7488242 TI - Biotransformation of all-trans-retinol and all-trans-retinal to all-trans retinoic acid in rat conceptal homogenates. AB - Catalysis of the oxidation of all-trans-retinol (vitamin A1) or of all-trans retinal to all-trans-retinoic acid (all-trans-RA) by rat conceptal enzymes was investigated during organogenesis. Products of the reaction were identified and quantified with HPLC by comparing their elution times with those of authentic standard retinoids. Under the incubation and assay conditions utilized, all-trans retinol and all-trans-retinal were converted to readily detectable quantities of all-trans-RA. Rat conceptal homogenates from gestational days 10.5, 11.5 and 12.5 each exhibited enzymatic activity for oxidation of all-trans-retinol and all trans-retinal to all-trans-RA. Enzymatic catalysis was verified by showing that: (1) both reactions were coenzyme dependent; (2) the rates of reactions increased as concentrations of conceptal protein increased; (3) both reactions were abolished by heating the tissue homogenates (100 degrees, 5 min); and (4) both reactions exhibited substrate saturation. Under the same experimental conditions, formation of all-trans-RA from all-trans-retinol was much slower than from all trans-retinal, suggesting that oxidation of all-trans-retinol to all-trans retinal was the rate-limiting step for biotransformation of all-trans-retinol to all-trans-RA in embryonic tissues. When NAD or NADP were replaced by NADH or NADPH, the rate of oxidation of all-trans-retinol was reduced markedly, indicating that the reaction was catalyzed primarily by an NAD/NADP-dependent dehydrogenase(s). Carbon monoxide (CO:O2 = 90:10) did not inhibit the reaction. NAD appeared to be a more effective cofactor than NADP in catalyzing oxidation of all-trans-retinal to all-trans-RA. When NAD was omitted, formation of all-trans RA from all-trans-retinal was reduced by approximately 55%. Replacing NAD by NADH or NADPH also reduced the conversion of all-trans-retinal to all-trans-RA by about 60%. These observations suggested at least two pathways for the generation of all-trans-RA from all-trans-retinal in embryos: oxidation catalyzed by an NAD/NADP-dependent dehydrogenase(s) and oxidation catalyzed by an oxidase(s) that did not require NAD, NADH, NADP or NADPH. Conversion of all-trans-retinol to all trans-RA was inhibited strongly by low concentrations of citral, but not by high concentrations of sodium azide, 4-methylpyrazole, or metyrapone. Similarly, oxidation of all-trans-retinal was inhibited strongly by citral but not by metyrapone.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 7488244 TI - Cytostatic activity of phenylacetate and derivatives against tumor cells. Correlation with lipophilicity and inhibition of protein prenylation. AB - The aromatic fatty acid phenylacetate, a common metabolite of phenylalanine, shows promise as a relatively non-toxic drug for cancer treatment. This slowly metabolized fatty acid alters tumor cell lipid metabolism causing, among other effects, inhibition of protein prenylation critical to malignant growth. In pursuit of more potent analogues, we have examined the activity of related compounds against tumor cell lines established from patients with advanced prostatic carcinoma, glioblastomas, and malignant melanoma. Like phenylacetate, derivatives containing alpha-carbon or ring substitutions induced cytostasis and phenotypic reversion at non-toxic concentrations. Potency was correlated with the degree of calculated lipophilicity of the aromatic fatty acid, and the extent of inhibition of protein prenylation. Remarkably, a parallel cytostatic activity was reported in embryonic plant cells, which respond to phenylacetate and its analogues in the same concentration range and the same rank order of lipophilicity. These data suggest that phenylacetate and its analogues may act through common mechanisms to inhibit the growth of vastly divergent, undifferentiated cell types, and provide a basis for the development of new agents for the treatment of human malignancies. PMID- 7488243 TI - Debenzoylating and deacetylating activities of rat liver and mammary gland microsomes. Effect of ovariectomy. AB - This study compared the rates of N-deacylations of N-hydroxy-N-2 fluorenylbenzamide (N-OH-2-FBA) with those of its analogue, N-hydroxy-N-2 fluorenylacetamide (N-OH-2-FAA), by the mammary gland (tumor target for both compounds) and the liver of female Sprague-Dawley rats and examined the effect of ovariectomy on these activities. N-Debenzoylation of N-OH-2-FBA was catalyzed by the mammary and liver microsomes of 50-day-old female rats at similar rates (approximately 24 nmol/min/mg). The activity of both tissues increased (up to 1.8 times) after ovariectomy at 42, 32 and, especially, 22 days of age. The rapid hydrolysis appeared to be unique for the benzoyl group since N-OH-2-FAA was deacylated only approximately 0.05 and 0.004 times as fast by the liver and mammary microsomes, respectively, and these low rates were unaffected by ovariectomy. Since such substrate specificity would be of significance in the metabolism of xenobiotics and drug design, esterase activity and its sensitivity to ovariectomy at 22 days of age were examined with several acetylated and benzoylated substrates in the liver and mammary microsomes and compared with those of male liver. Tissues of rats of both sexes had a greater capacity to hydrolyze carboxyl esters than amides. Expect for N-2-fluorenylacetamide (2-FAA) and o-nitrophenylacetate (o-NPA), all substrates were hydrolyzed by liver microsomes of the male up to 3.9 times faster than by those of the female. Microsomes of female liver hydrolyzed acetylated substrates 1.2 to 25 times faster than benzoylated analogues except for N-OH-2-FBA and benzamide. By contrast, mammary gland microsomes hydrolyzed benzoylated compounds 1.4 to 333 times faster except for 2-naphthyl benzoate. Respective rates of hydrolysis of o NPA by microsomes of liver and mammary gland were 1.7 and 0.6 times those of p NPA. After ovariectomy, deacylating activities increased (up to 1.6 times) except for those of 2-FAA and acetanilide. All deacylations were > 98% inhibited by 0.1 mM paraoxon, indicating catalysis by serine hydrolases. The results suggest involvement of multiple carboxylesterases and indicate that certain benzoylated xenobiotics may have a greater effect on the mammary gland than acetylated xenobiotics because of their greater vulnerability to hydrolysis by esterases of mammary gland. PMID- 7488245 TI - Induction of DT-diaphorase by doxorubicin and combination therapy with mitomycin C in vitro. AB - Mitomycin C (MMC) is a bioreductive antitumor agent that is activated by NADPH:cytochrome P450 reductase (EC 1.6.2.4) and NAD(P)H:(quinone acceptor) oxidoreductase (EC 1.6.99.2) (DT-diaphorase). DT-diaphorase is a two-electron reducing enzyme that is induced by a variety of chemicals, including quinones. Doxorubicin (DOX) is an anthraquinone antitumor agent that has been used clinically with MMC for combination chemotherapy in breast cancer. In this study, we investigated whether DOX could selectively induce DT-diaphorase in tumor cells and whether combining this agent with MMC in an appropriate schedule could produce synergistic antitumor activity. Treatment of EMT6 murine mammary tumor cells with DOX resulted in a 40% increase in DT-diaphorase activity in these cells, but had no effect on this enzyme in murine bone marrow cells. Combination therapy with DOX and MMC produced a 1.4-fold level of synergistic cell kill in the tumor cells, but a similar level of synergy was also observed in normal bone marrow cells. Thus, DOX can selectively induce elevated levels of DT-diaphorase in tumor cells; however, the synergy observed by combining this agent with MMC appears to be unrelated to the induction of DT-diaphorase. PMID- 7488246 TI - Comparison of transport properties of the reduced folate carrier and folate receptor in murine L1210 leukemia cells. AB - This laboratory previously described an L1210 murine leukemia cell line with a functional defect in the reduced folate carrier and increased expression of folate receptor-beta (F2-MTXrA). This cell line was used to characterize methotrexate (MTX) influx mediated by folate receptor-beta and to compare this with influx mediated by the reduced folate carrier in L1210 parental cells. Influx of 0.2 microM MTX in F2-MTXrA cells was one-third that of L1210 cells and was abolished by very low concentrations of folic acid. Kinetic analysis revealed that MTX transport mediated by folate receptor-beta exhibited an influx kappa t one-third, and an influx Vmax one-fourth, that of the reduced folate carrier. Metabolic inhibitors markedly suppressed influx in F2-MTXrA cells but had no effect on MTX influx in L1210 cells. MTX influx in both cell lines was inhibited by the organic anions probenecid, sulfobromophthalein, and CI-920, but to a lesser extent in F2-MTXrA cells. The inhibitory effects of these anions on transport in F2-MTXrA cells could be attributed to their inhibition of MTX binding to the folate receptor. Although MTX influx in both cell lines was not sodium dependent, removal of extracellular chloride increased influx 2-fold in L1210 cells while markedly inhibiting influx in F2-MTXrA cells. Substitution of Cl- with isethionate or NO3- partially restored influx in the latter cells, whereas SO4(2-) was inhibitory. Anions enhanced MTX binding to folate receptor beta with isethionate > SO4(2-) > Cl-. Decreasing the buffer pH to 6.2 produced a 69% reduction, and a 260% increase, in MTX influx in L1210 cells and F2-MTXrA cells, respectively. The data indicate that folate receptor-beta-mediated MTX influx has properties fundamentally different from transport mediated by the reduced folate carrier in terms of energy, ion, and pH dependence. There was no evidence indicating that these processes are functionally linked. PMID- 7488248 TI - Effect of genistein on both basal and glucagon-induced levels of cAMP in rat hepatocytes. AB - Using rat liver hepatocytes, we studied the effect of the tyrosine kinase inhibitor genistein on the Ca2+/IP3 (inositol 1,4,5-trisphosphate) and the cAMP (adenosine 3:5-cyclic monophosphate) transduction mechanisms. Genistein specifically blocked the activation of glycogen phosphorylase after EGF (epidermal growth factor). Genistein on its own partially activated phosphorylase and inactivated glycogen synthase. Genistein did not influence levels of IP3, but increased those of cAMP. This was especially clear when genistein was given together with glucagon. The data suggest an effect of a tyrosine kinase on the synthesis/degradation of cAMP. PMID- 7488247 TI - Functional analysis of aryl hydrocarbon receptor nuclear translocator interactions with aryl hydrocarbon receptor in the yeast two-hybrid system. AB - The aryl hydrocarbon receptor (AHR) mediates dioxin (2,3,7,8-tetrachlorodibenzo-p dioxin)-induced transcriptional activation of a battery of genes by interaction with a cofactor, called aryl hydrocarbon receptor nuclear translocator (ARNT) protein. Both AHR and ARNT belong to a family of proteins that includes the Drosophila circadian-rhythm protein and "single-minded" protein. These proteins share a domain called the PAS domain. In addition to the PAS domain, both AHR and ARNT contain basic helix-loop-helix (bHLH) and glutamine (Q)-rich domains. The roles of these domains in the receptor-mediated transcriptional activation are not understood completely. By using the yeast two-hybrid system with the N terminal half of AHR as a probe, which contains the bHLH and PAS regions, to screen cDNA libraries prepared from human lymphocytes and C57BL mouse liver for clones encoding proteins capable of binding to these regions, we isolated a partial ARNT cDNA clone. These results demonstrated that the N-terminal half of AHR is capable of interacting with ARNT in yeast (probably through the bHLH motif). A fusion protein containing the GAL4 DNA binding domain (DB) linked to the full-length AHR was not capable of activating expression of a reporter gene containing the GAL4 DNA binding site, suggesting that ligand-free AHR alone has no transactivating properties in yeast. However, the C-terminal portion (amino acid residues 580-797) of the AHR, including the Q-rich domain, could confer transactivation of the reporter gene expression in the same system, suggesting that the N-terminal portion of the AHR contains transcription repression properties. In contrast, GAL4(DB)-ARNT fusion protein was able to activate expression of the same reporter gene. Deletion analysis of ARNT revealed that the C-terminal 75 amino acids, including the Q-rich domain, exhibited full transactivation function in yeast and mammalian cells. These results revealed different structural organizations for the transactivation properties between AHR and ARNT, although both contained transactivation domains at the C-termini. PMID- 7488249 TI - Persistence of long-lasting serotonin depletion by p-chloroamphetamine in rat brain after 6-hydroxydopamine lesioning of dopamine neurons. AB - In rats that had been treated neonatally with 6-hydroxydopamine (6OHDA) to deplete striatal dopamine more than 95%, a single injection of p chloroamphetamine (pCA) (5 or 10 mg/kg, i.p.) resulted in depletion of striatal and hippocampal serotonin at 1 week to a similar extent as in control rats. These findings suggest that striatal dopamine is not essential to the long-lasting depletion of brain serotonin by pCA in rats. PMID- 7488250 TI - Effect of food intake on the activity of liver enzymes in partially hepatectomized rats treated with tumor necrosis factor. AB - After partial hepatectomy (PHx), there are significant changes in the activity of a number of enzymes in the regenerating rat liver. Administration of low doses of recombinant human tumor necrosis factor-alpha (rHu-TNF) to normal rats induces similar changes in some of the enzymes but not in others. Because certain observations suggest that TNF may play a dominant role in liver regeneration, we speculated that the discrepancies in enzyme activities may be due to the decrease in food intake caused by PHx. Accordingly, the activities of eleven liver enzymes of 70% PHx rats additionally treated i.p. with rHu-TNF (20-50 micrograms/kg/day for 3-4 days) were compared with those of (i) PHx controls fed ad lib., and (ii) PHx controls pair-fed the same amount of food. When pair-fed controls were used, the discrepancies in the activities of the enzymes that are affected by fasting tended to disappear, suggesting that the decrease in the food intake was responsible for the differences. PMID- 7488251 TI - Scavenging effect of silipide, a new silybin-phospholipid complex, on ethanol derived free radicals. AB - Ethanol metabolism by cytochrome P4502E1 (CYP2E1) produces free radical intermediates, identified as hydroxyethyl radicals. We have observed that in vitro addition or in vivo pretreatment of rats with Silipide, a new 1:1 complex of silybin with phosphatidyl-choline, is able to decrease the spin trapping of hydroxyethyl radicals in microsomes from chronic alcohol-fed rats. This effect is not due to an interference with the metabolism of ethanol by CYP2E1, but is rather related to the capacity of the silybin molecule to scavenge hydroxyethyl radicals. However, such an effect is lost when pure silybin in amounts comparable to those present in Silipide is administered instead, due to the low bioavailability of uncomplexed flavonoid. Further experiments in vivo have shown that Silipide administration also decreases hydroxyethyl radical signals detectable in the bile of rats acutely treated with ethanol. The ability of Silipide to scavenge ethanol-derived radicals along with its antioxidant activity suggests that this drug might be potentially useful in counteracting free radical mediated injuries involved in the development of liver damage caused by alcohol abuse. PMID- 7488252 TI - Selective damage to the active X chromosome by camptothecin and amsacrine as determined by an allele-specific alkaline unwinding assay. AB - Previous studies with MCF-7 cells demonstrated that several agents induce greater strand breakage in active genes than in nontranscribed centromeric regions. To better assess the effects of gene activity and inactivity, an allele-specific DNA strand break assay was developed, which allowed direct comparison of damage at a specific genetic locus on the active and inactive X chromosomes. The ZP lymphoblastoid cell line is heterozygous at the glucose-6-phosphate dehydrogenase (G6PD) locus, and the unexpressed (A) allele on the inactive X chromosome contains a FokI restriction site that is lacking in the expressed (B) allele on the active X. ZP cells were treated with camptothecin or amsacrine, and subjected to alkaline-induced DNA unwinding. Following detergent lysis and digestion of single-stranded DNA with S1 nuclease, the remaining double-stranded DNA was isolated and subjected to polymerase chain reaction (PCR) with primers that flank the polymorphic FokI site, with [alpha-32P]dCTP being added in the last PCR cycle. The resulting labeled PCR product was cleaved with FokI to assess the A/B allele ratio in the double-stranded DNA fraction. Treatment with camptothecin and amsacrine increased the apparent A/B ratio by factors of 2-3 and 1.5-2 respectively, indicating that the active B allele is preferentially damaged by these agents. PMID- 7488253 TI - Evaluation of whole-animal data using the ion parametric resonance model. AB - Changes observed in the behavioral response of land snails from exposure to parallel ac and dc magnetic fields demonstrate limited agreement with the predictions of an interaction model proposed by Lednev and the predictions of a recently proposed ion parametric resonance (IPR) model. However, the inadequate number of reported data points, particularly in a critical exposure range, prevents unambiguous application of either the Lednev or the IPR model. PMID- 7488254 TI - Magnetic field exposure among utility workers. AB - The Electric and Magnetic Field Measurement Project for Utilities--the Electric Power Research Institute (EPRI) Electric and Magnetic Field Digital Exposure (EMDEX) Project (the EPRI EMDEX Project)--was a multifaceted project that entailed technology transfer, measurement protocol design, data management, and exposure assessment analyses. This paper addresses one specific objective of the project: the collection, analysis, and documentation of power-frequency magnetic field exposures for a diverse population of utility workers. Field exposure data measured by an EMDEX system were collected by volunteer utility employees at 59 sites in four countries between September, 1988, and September, 1989. Specially designed sampling procedures and data collection protocols were used to ensure uniform implementation across sites. Volunteers within 13 job classifications recorded which of eight work or three nonwork environments they occupied while wearing an EMDEX meter. Approximately 50,000 hours of magnetic field exposure records taken at 10 s intervals were obtained, about 70% of which were from work environments. Exposures and time spent in environments were analyzed by primary work environment, by occupied environment, and by job classification. Generally, for utility-specific job classifications related to the generation, transmission, and distribution of electricity, the field and exposure measurements in terms of workday mean field were higher than in more general occupations. The job classifications with the highest (median workday mean) exposure were substation operators (0.7 microT) and electricians (0.5 microT). Total variance also tended to be largest for utility-specific job classifications. For these workers, the contributions of between-worker and within-worker variances to total variance were about the same. Measurements in utility-specific environments were higher than in more general environments. Estimates of time-integrated exposure indicated that utility-specific job classifications received about one-half or more of their total exposure on the job. The nonwork field and exposure distributions for workers in all job categories were comparable with median nonworkday means of about 0.09 microT. PMID- 7488255 TI - Power lines and the geomagnetic field. AB - The metric of prime interest in power line epidemiological studies has been AC magnetic intensity. To consider also possible geomagnetic involvement, the orientation of a long straight power line is examined relative to a uniform geomagnetic field (GMF) with dip angle alpha. An expression is derived for the component of the total GMF that is parallel, at an elevation beta, to the circuital magnetic field that surrounds the line. This component is a function of the angles alpha and beta, the total geomagnetic intensity BT, and the angle theta between the axis of the power line and magnetic north. Plotting these geomagnetic parameters for known leukemia residences allows one to test for possible ion cyclotron resonance or other GMF interactions. This approach, in principle, is an easy addition to existing or planned studies, because residential access is not required to obtain local values for alpha, beta, theta, and BT. We recommend including these parameters in the design of epidemiological studies examining power line fields and childhood leukemia. PMID- 7488256 TI - Applied AC and DC magnetic fields cause alterations in the mitotic cycle of early sea urchin embryos. AB - This study demonstrates that exposure to 60 Hz magnetic fields (3.4-8.8 mT) and magnetic fields over the range DC-600 kHz (2.5-6.5 mT) can alter the early embryonic development of sea urchin embryos by inducing alterations in the timing of the cell cycle. Batches of fertilized eggs were exposed to the fields produced by a coil system. Samples of the continuous cultures were taken and scored for cell division. The times of both the first and second cell divisions were advanced by ELF AC fields and by static fields. The magnitude of the 60 Hz effect appears proportional to the field strength over the range tested. The relationship to field frequency was nonlinear and complex. For certain frequencies above the ELF range, the exposure resulted in a delay of the onset of mitosis. The advance of mitosis was also dependent on the duration of exposure and on the timing of exposure relative to fertilization. PMID- 7488257 TI - Microwave influence on the isolated heart function: I. Effect of modulation. AB - Dependence of the microwave effect on modulation parameters (pulse width, duty ratio, and peak intensity) was studied in an isolated frog auricle preparation. The rate and amplitude of spontaneous auricle twitches were measured during and after a 2 min exposure to 915 or 885 MHz microwaves and were compared to preexposure values. The studied ranges of modulation parameters were: pulse width, 10(-6)-10(-2) s; duty ratio, 7:100000, and peak specific absorption rate, 100-3000 W/kg. Combinations of the parameters were chosen by chance, and about 400 various exposure regimes were tested. The experiments established that no regime was effective unless the average microwave power was high enough to induce preparation heating (0.1-0.4 degree C). The twitch rate instantly increased, and the amplitude decreased, as the temperature rose; similar changes could be induced by equivalent conventional heating. The data provide evidence that the effect of short-term microwave exposure on the isolated heart pacemaker and contractile functions depends on pulse modulation just as much as modulation determines the average absorbed power. These functions demonstrated no specific dependence on exposure parameters such as frequency or power windows. PMID- 7488258 TI - Microwave influence on the isolated heart function: II. Combined effect of radiation and some drugs. AB - The combined effects of microwave radiation and some drugs were studied in an isolated frog auricle preparation. The experiments established that exposure to pulse-modulated 915 MHz microwaves for up to 40 min had no effect on either the rate or the amplitude of spontaneous auricle twitches, unless the average absorbed power was high enough to produce preparation heating. Treatment of the preparation with saline containing (0.6-3.0) 10(-5) M of propranolol or (0.5-1.5) 10(-7) M of atropine altered neither its pacemaker nor its contractile functions; these drugs also had no effect when they were combined with nonthermal microwave irradiation. Caffeine (1 mM) strongly increased the average heart power, which was calculated as the product of twitch rate and amplitude. The caffeine effect appeared to be significantly augmented (by about 15%, P < 0.02) under exposure to burst-type pulsed microwaves (pulse width, 1.5 msec; pause, 2.5 msec; 8 pulses/burst, 16 bursts/s; average SAR, 8-10 W/kg). By itself, this modulation was not effective; the heating of the preparation and saline during exposure was approximately 0.1 degrees C, which could not account for the detected changes. The experimental results demonstrate that caffeine treatment increases the microwave sensitivity of the frog auricle preparation and reveals primarily subthreshold, nonthermal microwave effect. PMID- 7488259 TI - Membrane potential perturbations induced in tissue cells by pulsed electric fields. AB - Pulsed electric fields directly influence the electrophysiology of tissue cells by transiently perturbing their transmembrane potential. To determine the magnitude and time course of this interaction, electrotonic cable theory was used to calculate the membrane potential perturbations induced in tissue cells by a spatially uniform, pulsed electric field. Analytic solutions were obtained that predict shifts in membrane potential along the length of cells as a function of time in response to an electrical pulse. For elongated tissue cells, or groups of tissue cells that are coupled electrotonically by gap junctions, significant hyperpolarizations and depolarizations can result from millisecond applications of electric fields with strengths on the order of 10-100 mV/cm. The results illustrate the importance of considering cellular cable parameters in assessing the effects of transient electric fields on biological systems, as well as in predicting the efficacy of pulsed electric fields in medical treatments. PMID- 7488260 TI - Neurochemical effects of a 20 kHz magnetic field on the central nervous system in prenatally exposed mice. AB - C57/B1 mice were exposed during pregnancy (gestation days 0-19) to a 20 kHz magnetic field (MF). The asymmetric sawtooth-waveform magnetic field in the exposed racks had a flux density of 15 microT (peak to peak). After 19 days, the exposure was terminated, and the mice were housed individually under normal laboratory conditions. On postnatal day (PD) 1, PD21, and PD308, various neurochemical markers in the brains of the offspring were investigated and the brains weighed. No significant difference was found in the whole brain weight at PD1 or PD21 between exposed offspring and control animals. However, on PD308, a significant decrease in weight of the whole brain was detected in exposed animals. No significant differences were found in the weight of cortex, hippocampus, septum, or cerebellum on any of the sampling occasions, nor were any significant differences detected in protein-, DNA-level, nerve growth factor (NGF), acetylcholine esterase- (AChE), or 2',3'-cyclic nucleotide 3' phosphodiesterase- (CNP; marker for oligodendrocytes) activities on PD21 in cerebellum. Cortex showed a more complex pattern of response to MF: MF treatment resulted in a decrease in DNA level and increases in the activities of CNP, AChE, and NGF protein. On PD308, the amount of DNA was significantly reduced in MF treated cerebellum and CNP activity was still enhanced in MF-treated cortex compared to controls. Most of the effects of MF treatment during the embryonic period were similar to those induced by ionizing radiation but much weaker. However, the duration of the exposure required to elucidate the response of different markers to MF seems to be greater and effects appear later during development compared to responses to ionizing radiation. PMID- 7488261 TI - Comments on "Clarification and application of ion parametric resonance model for magnetic field interactions with biological systems" by Blanchard and Blackman. PMID- 7488262 TI - [Structural analysis of isoforms of Na+,K+-ATPase from calf brain]. AB - The isoform composition and types of functioning of Na+,K(+)-ATPase complexes, as well as their ouabain-inhibition constants, were studied for calf brain membranes. The catalytic subunit alpha 3 within the native enzyme complex was found to exhibit an increased sensitivity to endogenous proteolysis. The site of specific proteolysis was localized in the region of the polypeptide chain that is unique for all alpha 3 type isoforms: PNDNR492 decreases (Y493) (according to the numeration of human alpha 3-subunit). It was shown for the first time that in all enzyme preparations containing the alpha 2 and alpha 3 isoforms isolated by both Jorgensen's and Esmann's method two other proteins were present: the beta 5 chain of tubulin and glyceraldehyde-3-phosphate dehydrogenase; the biological meaning of their association is still unclear. PMID- 7488263 TI - [Structural properties of ribosomal protein S8 from the extreme thermophile Thermus thermophilus]. AB - The gene of ribosomal protein S8 from the extreme thermophile Thermus thermophilus was expressed in E. coli using the strain BL21(DE3) and vector pET3 1. A method of isolating this protein from the super producing strain was developed, which makes it possible to obtain 8-12 mg of product from 11 of culture. The secondary structure of protein S8 was determined by using CD spectroscopy. The protein was shown to be highly resistant to denaturants. PMID- 7488264 TI - [The automated "SynChrom" system for solid-phase synthesis of peptides and liquid column chromatography. I. Principles of design and structural constituents]. AB - An automatic modular SynChrom system was designed for solid phase synthesis and chromatographic purification of peptides. Structural constituents and organization and operation of the system in solid phase peptide synthesis and liquid column chromatography modes are described. Swellographic monitoring of the course of synthesis was used. Hydraulic diagrams, operation algorithm and software description are presented. PMID- 7488265 TI - [The automated "SynChrom" system for solid-phase synthesis of peptides and liquid column chromatography. II. Use in the solid-phase synthesis of peptides and liquid column chromatography]. AB - Automatic solid phase peptide synthesis using the SynChrom system is described. Problems of swellographic monitoring are discussed. Combined monitoring (swellographic, spectrophotometric, and manometric) of all steps of the synthetic cycle are suggested. Potential applications of the system to liquid column chromatography were demonstrated. PMID- 7488266 TI - [Synthesis of octadecyl esters of hydrophobic proline-containing peptides]. PMID- 7488267 TI - [Cloning and expression of the Escherichia coli thioredoxin gene]. AB - The gene encoding thioredoxin (trxA) was isolated from chromosomal DNA of E. coli HB101 strain using the polymerase chain reaction. The cloned structural gene with a synthetic Shine-Dalgarno sequence was placed under the control of either inducible tac-promoter or a tandem of two strong constitutive promoters A2 and A3 from early region of bacteriophage T7. Both constructions were shown to provide high levels of biosynthesis of recombinant thioredoxin. PMID- 7488268 TI - [A full-size DNA copy of the tick-borne encephalitis virus genome. I. Analysis of the 5'- and 3'-terminal noncoding regions of the genome]. AB - Using reverse transcription and the polymerase chain reaction, cDNA fragments of noncoding regions of the tick-borne encephalitis virus (TBEV) genome were obtained. These fragments were cloned into a pGEM3 vector, and their nucleotide sequences were determined. The heterogeneity of the 3'-terminal untranslated region of the TBEV RNA was revealed. To create a stable full-size DNA copy of the TBEV genome, four cDNA variants differing in length and structure of the 3' terminal fragment of the viral RNA were cloned into a pBR322-derived vector. PMID- 7488269 TI - [A new approach to the problem of immobilizing oligonucleotides on carboxyl containing nylon membranes for nucleic acid hybridization]. AB - A simple and efficient method of the covalent immobilization of oligonucleotides on carboxyl-containing nylon membranes was proposed. The method is based on the reaction between a postsynthetically introduced aminoalkyl group of an oligonucleotide and membrane carboxyl catalyzed by a water soluble N-(3 dimethylaminopropyl)-N'-ethylcarbodiimide. Judging from the mechanism of carboxyl group activation by the carbodiimide, optimal conditions of the immobilization were selected, which made it possible to increase the immobilization efficiency and carrier capacity as compared with those described previously. Using a 23-unit membrane-immobilized oligonucleotide as an example, the oligonucleotides immobilized were shown to be capable of hybridizing with complementary sequences. PMID- 7488270 TI - [Synthesis and some biochemical properties of phosphonyl acyclic analogs of 2' deoxyadenosine nucleotides]. AB - 9-[2-(phosphonomethylcarbonylamino)ethyl]adenine, 9-[(2 phosphonoethyl)aminocarbonylmethyl]adenine, 9-[2-[(2 phosphonoethyl)carbonylamino]ethyl]adenine, and their diphosphates were synthesized. All three diphosphates were shown to incorporate into the 3' terminus of the DNA chain during the synthesis of the avian myeloblastosis virus catalyzed by reverse transcriptase. However, they do not serve as substrates for DNA polymerase alpha from human placenta, polymerase beta from calf thymus, or terminal deoxynucleotidyl transferase from calf thymus. PMID- 7488271 TI - Some guidelines about practice guidelines. PMID- 7488272 TI - Guidelines for the medical management of osteoarthritis. Part I. Osteoarthritis of the hip. American College of Rheumatology. AB - Treatment of patients with OA of the hip should be individualized and tailored to the severity of the disease. In individuals with mildly symptomatic disease, treatment may be limited to patient education, physical and occupational therapy, other nonpharmacologic modalities, and drug therapy with a non-opioid oral analgesic. In patients who are unresponsive to this treatment regimen, the use of an NSAID in addition to nonpharmacologic therapy is appropriate unless it is medically contraindicated. Patients with severe symptomatic OA of the hip require an aggressive approach to decreasing pain, increasing mobility, and improving function; such patients may benefit from orthopedic consultation and evaluation for osteotomy or total joint arthroplasty. PMID- 7488273 TI - Guidelines for the medical management of osteoarthritis. Part II. Osteoarthritis of the knee. American College of Rheumatology. AB - Treatment of patients with OA of the knee should be individualized and tailored to the severity of the symptoms. In individuals with mild symptomatic OA, treatment may be limited to patient education, physical and occupational therapy and other nonpharmacologic modalities, and pharmacologic therapy including non opioid oral and topical analgesics. In patients who are unresponsive to this treatment regimen, the use of NSAIDs in addition to nonpharmacologic therapy is appropriate unless medically contraindicated. Judicious use of intraarticular steroid injections has a role either as monotherapy or an adjunct to systemic therapy in patients with knee OA who have symptomatic effusions. The role of joint lavage and arthroscopic debridement in patients with OA of the knee who are unresponsive to conservative medical therapy needs further study, and these procedures cannot be routinely recommended for all patients at this time. Patients with severe symptomatic OA of the knee require an aggressive approach to decreasing pain, increasing mobility, and decreasing functional impairment; such patients may benefit from orthopedic consultation and evaluation for osteotomy or total joint arthroplasty. PMID- 7488274 TI - Pathogenesis of spondylarthropathies. Persistent bacterial antigen, autoimmunity, or both? AB - We have discussed partially mutually exclusive, partially overlapping models for the pathogenesis of the spondylarthropathies. Not all possibilities have been presented here; others are discussed elsewhere (77, 78). Furthermore, we have not addressed the issue of B27-negative spondylarthropathy. However, in our opinion, the key to understanding the pathogenesis of the spondylarthropathies lies in the interaction between the class I MHC molecule HLA-B27 and the T cell response. Although a T cell response driven by persisting bacterial antigen is still an attractive hypothesis, it does not explain all the known aspects of spondylarthropathy pathogenesis. The possibility of autoimmunity triggered by bacterial infection needs also to be considered, especially the new idea of HLA B27-derived peptides presented by class II MHC molecules. The predominant involvement of joints is not easily explained in the case of autoimmunity. Cross reactivity to joint-specific structures such as type II collagen (79) and/or bacteria inside the joint at the beginning of the immune response, with induction of local autoimmunity, might be involved. Most of the issues raised here could be tested by experiment, and we can expect to learn soon whether any of these models will explain the pathogenesis, or if we have to look further. The PCR technique will facilitate the search for bacteria not only in peripheral joints, but also now in sacroiliac biopsy samples from patients with AS and other spondylarthropathies. A prospective study on ReA in an endemic area should teach us more about predisposing factors (for example for Shigella-induced enteritis, which occurs in many parts of the world outside Europe and the US) (80).(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7488275 TI - Evaluative laboratory testing. Assessing tests that assess disease activity. PMID- 7488276 TI - The elusive search for geographic clusters of systemic lupus erythematosus. Critical review. AB - OBJECTIVE: To ascertain whether there are any geographic clusters of systemic lupus erythematosus (SLE), and critically review 2 previous reports of these clusters. METHODS: A literature review of all epidemiologic surveys relevant to SLE published since 1966 was undertaken. This included a search of abstracts and reports, in addition to peer-reviewed publications. RESULTS: Two geographic clusters were identified in the literature. A report purporting that contaminated ground water in southwest Tucson, Arizona was associated with significantly increased symptoms of SLE was flawed since no patients were examined, discarded criteria were used, and incorrect definitions were employed by the nonrheumatologist authors. Another study, which identified a cluster in Nogales, Arizona, used generally valid methodology. However, the authors' conclusion that a 94/100,000 prevalence of SLE was evidence of an excessive increase in this 92% Hispanic/Native American community is not consistent with published surveys among Hispanic and Native American populations. CONCLUSION: No valid claims of SLE geographic clusters have been published. PMID- 7488277 TI - Which outcome measures should be used in rheumatoid arthritis clinical trials? Clinical and quality-of-life measures' responsiveness to treatment in a randomized controlled trial. AB - OBJECTIVE: To determine the discriminant validity of the core set of outcome measures proposed by the American College of Rheumatology (ACR) and the Outcome Measures in Clinical Trials (OMERACT) conference committee to be used in clinical trials of rheumatoid arthritis (RA). METHODS: Utilizing data from a multicenter randomized double-blind clinical trial of low-dose cyclosporine and placebo in RA, we estimated the relative efficiency (RE) of measures to detect a treatment effect (relative to tender joint count, which was assigned a value of 1). Four pain measures (10-cm visual analog scale [VAS], 5-point categorical scale, Health Assessment Questionnaire [HAQ] pain index, Arthritis Impact Measurement Scales [AIMS] pain score) and 3 quality-of-life measures (Problem Elicitation Technique [PET], HAQ, AIMS) were compared. RESULTS: Physician and patient global measures were the most responsive instruments, although neither was statistically superior to tender joint count. Swollen joint count, grip strength, pain measured on a 10 cm VAS, and functional status as measured by the PET and HAQ were all of intermediate responsiveness. Morning stiffness, 5-point pain scale, and erythrocyte sedimentation rate were the least responsive instruments. CONCLUSION: This study provides further evidence to support the core set of outcome measures proposed by the ACR and OMERACT: PMID- 7488278 TI - Double-blind, placebo-controlled multicenter trial using chimeric monoclonal anti CD4 antibody, cM-T412, in rheumatoid arthritis patients receiving concomitant methotrexate. AB - OBJECTIVE: To evaluate the clinical response to and safety of single and repeat doses of a chimeric anti-CD4 monoclonal antibody, cM-T412, in patients with rheumatoid arthritis (RA) concomitantly treated with a stable regimen of low-dose methotrexate. METHODS: Sixty-four patients with refractory RA, who were already receiving stable doses of methotrexate, were randomized into a multicenter, double-blind, placebo-controlled trial to receive 3 monthly treatments with either a placebo, or 5, 10, or 50 mg cM-T412, given intravenously. RESULTS: Using > or = 50% improvement in swollen joint counts as a criterion for clinical response, 13%, 13%, 18%, and 13% of patients receiving 50, 10, or 5 mg cM-T412, or the placebo, respectively, exhibited a clinical response at 3 months of therapy. Using > or = 50% improvement in tender joint counts as a measure of clinical efficacy at 3 months, 19%, 13%, 12%, and 6% of patients receiving 50, 10, or 5 mg cM-T412, or the placebo, respectively, exhibited a clinical response. "Flu-like" symptoms (fever, chills, rigor) within 24 hours of the infusion occurred more frequently in the groups receiving 50-mg (29%) and 10-mg (31%) doses of cM-T412 than those receiving 5 mg cM-T412 (12%) or the placebo (13%). Significant CD4+ T cell depletion occurred in the 50-mg group (mean of 353 CD4+ T cells/mm3 at 6 months versus 856 CD4+ T cells/mm3 at baseline). All patients were followed up for 12 months after the final treatment; no opportunistic infectious complications occurred. CONCLUSION: Treatment with cM-T412 in this cohort of RA patients who were also taking methotrexate was not associated with clinical efficacy or enhanced toxicity from infectious complications, despite significant peripheral CD4+ T cell depletion. PMID- 7488279 TI - CAMPATH-1H, a humanized monoclonal antibody, in refractory rheumatoid arthritis. An intravenous dose-escalation study. AB - OBJECTIVE: To evaluate the biologic response, tolerability, and potential clinical effect of a humanized antilymphocyte monoclonal antibody, CAMPATH-1H, in patients with rheumatoid arthritis (RA). METHODS: Forty adult patients with active, refractory RA were treated with CAMPATH-1H, given intravenously, in a multicenter, open, single-dose-escalation study. Patients were assigned to dose groups of 1, 3, 10, 30, 60, and 100 mg CAMPATH-1H. RESULTS: There was a profound, immediate, and sustained reduction of the peripheral lymphocyte count; the most susceptible were the levels of CD4+ and CD8+ cells, which remained depressed during the study period. Sixty-three percent of patients developed antibodies to CAMPATH-1H. Side effects occurred frequently throughout the first 24 hours following infusion, and included fever, headache, nausea, vomiting, and hypotension. All of the immediate drug toxicities resolved within the initial 24 hour postdosing period. One patient developed a reactivation of Mycobacterium xenopi infection 10 weeks following infusion. Sixty-five percent of patients developed a clinical response; the mean duration of response was 2 weeks. CONCLUSION: CAMPATH-1H is a lymphocyte-depleting antibody that is biologically potent even after single-dose therapy. There was no correlation between biologic effect and clinical response. Sustained lymphocyte suppression was observed. Acute infusion toxicities were observed in most patients. The role of depleting monoclonal antibodies in the treatment of RA should be reevaluated. PMID- 7488281 TI - Oral 2-chlorodeoxyadenosine in psoriatic arthritis. A preliminary report. AB - OBJECTIVE: To determine if weekly oral 2-chlorodeoxyadenosine (2-CdA) can induce selective lymphocytopenia, and reduce inflammation, in patients with refractory psoriatic arthritis. METHODS: Seven patients with psoriatic arthritis were treated with oral 2-CdA at weekly dosages of 0.3 mg/kg to 0.45 mg/kg for 12 weeks, followed by monthly maintenance therapy. The patients were evaluated after 6 months. RESULTS: The drug treatment produced selective lymphocytopenia, and reduced lymphocyte infiltration into involved skin. One patient did not complete 12 weeks of therapy because of perceived lack of efficacy. Four of the 6 remaining patients had improved joint disease, and 5 of 6 had improved psoriasis. CONCLUSION: Weekly oral 2-CdA appears to be a well-tolerated regimen for the inducement of peripheral lymphocytopenia in patients with psoriatic arthritis. Larger-scale, controlled trials may be warranted. PMID- 7488280 TI - Safety and effectiveness of leflunomide in the treatment of patients with active rheumatoid arthritis. Results of a randomized, placebo-controlled, phase II study. AB - OBJECTIVE: To assess the safety and effectiveness of leflunomide versus placebo in patients with active rheumatoid arthritis (RA) treated for 6 months. METHODS: Four hundred two patients were randomly assigned to receive placebo or leflunomide at 5 mg, 10 mg, or 25 mg daily. A washout period of 6-12 weeks from prior second-line therapy was required. RESULTS: Statistically significant improvement in primary and secondary outcome measures, as well as by responder analyses, occurred in the 10-mg and 25-mg dosage groups compared to placebo. Twenty-one patients (7.0%) in the active treatment groups withdrew due to adverse events (AEs). The incidence of AEs was higher with leflunomide than with placebo. Gastrointestinal symptoms, weight loss, allergic reactions, skin rash, and reversible alopecia were more common in the 10-mg and 25-mg dosage groups. The incidence of infections was similar between the treatment and placebo groups; no opportunistic infections were seen. Transient elevations in liver function studies were noted in a small number of patients. CONCLUSION: Leflunomide is effective in daily doses of 10 mg and 25 mg in patients with active RA. Improved efficacy at the 25-mg dose was associated with a higher incidence of AEs. Randomized, placebo-controlled trials using daily doses of 10 mg and 20 mg are under way in the US and Europe to confirm these positive results. PMID- 7488282 TI - Quantification of rheumatoid synovitis by magnetic resonance imaging. AB - OBJECTIVE: To develop a method for quantifying acute synovial inflammation in rheumatoid arthritis (RA), utilizing magnetic resonance imaging (MRI). METHODS: Gadolinium-diethylenetriamine pentaacetic acid-enhanced MRI was performed in 21 patients with knee synovitis. Changes in synovial membrane signal intensity were identified and quantified by line profile analysis. Multiple synovial biopsies were obtained by a blind biopsy technique, and standard clinical and laboratory measurements of disease activity were recorded. RESULTS: The rate of synovial membrane enhancement correlated with histologic features of acute inflammation (r = 0.63, P < 0.01), but not with clinical or laboratory assessments. CONCLUSION: Dynamic MRI is a valuable technique for assessing acute synovial inflammation in RA. PMID- 7488284 TI - Predictors of work disability in rheumatoid arthritis patients. A five-year followup. AB - OBJECTIVE: To evaluate regression models that include social, attitudinal, work structure, health status, and family characteristics, with regard to their prediction of work disability in a national sample of patients with rheumatoid arthritis (RA). METHODS: Four hundred ninety-eight employed RA patients were recruited from a national sample of private rheumatology practices. Three hundred ninety-two remained in the study after 5 years. Data were collected from patients by telephone interview, and patients' physicians provided written clinical assessments. Only variables on which information was obtained in year 1 were used to predict work status in year 5, using hierarchical multiple logistic regression analysis. RESULTS: The significant predictors of work disability were age (odds ratio [OR] 1.04), number of deformed joints (OR 1.26), number of joints with flare (OR 1.23), the complexity of working with things at work (OR 0.88), and the desire to remain employed (OR 2.3). The risk of work disability increased with increasing age, more severe disease, greater complexity of involvement with things at work, reduced work hours, and desire to not be working outside the home. CONCLUSION: The risk of becoming work disabled in 5 years was predicted more by clinical status at entry into the study than by work structure. These results, which contradict previous research on work disability in arthritis, prompt a rethinking of future studies of work disability in RA. PMID- 7488283 TI - Peripheral neuropathy with necrotizing vasculitis in rheumatoid arthritis. A clinicopathologic and prognostic study of thirty-two patients. AB - OBJECTIVE: To examine the clinicopathologic features of the noncompressive neuropathies in rheumatoid arthritis (RA). METHODS: We studied 32 patients with RA and peripheral neuropathy whose nerve and/or muscle biopsy specimens exhibited necrotizing vasculitis. Morphologic analysis of nerve specimens included light and electron microscopy studies and teased fiber preparation. Survival was evaluated, and the prognostic values of clinical, biologic, and pathologic features were assessed by Cox proportional hazards model. A prognostic assessment based on the significant variables was devised to estimate the probability of survival of any individual patient. RESULTS: Epi- and/or perineurial vasculitis was observed with the same frequency in the 17 patients with sensory and motor deficit and the 15 patients with sensory neuropathies and was associated with axonal degeneration of an average of 77.7% of the nerve fibers. The mean followup was 7.2 years, and the overall survival rate at 5 years was 57%. A full prolonged remission of the vasculitis was observed in 53% of the patients; relapse occurred in 25%. The factors correlated with mortality, in decreasing order of significance, were clinical cutaneous vasculitis (P = 0.0003), neuropathy affecting 3 or 4 limbs (P = 0.03), and depressed level of C4 (P < 0.05). The prognostic assessment indicated a wide range of 5-year probabilities of survival, from < 1% to 93%. CONCLUSION: Necrotizing vasculitis is responsible for the different patterns of noncompressive neuropathies in RA, including mononeuritis multiplex and distal symmetric sensory or sensorimotor neuropathy. Cutaneous vasculitis, multifocal neuropathy, and depressed C4 level were the 3 independent variables which best predicted mortality. We propose a prognostic assessment according to these variables, to stratify patients to receive more aggressive or less aggressive therapy. PMID- 7488285 TI - Corticosteroids plus pulse cyclophosphamide and plasma exchanges versus corticosteroids plus pulse cyclophosphamide alone in the treatment of polyarteritis nodosa and Churg-Strauss syndrome patients with factors predicting poor prognosis. A prospective, randomized trial in sixty-two patients. AB - OBJECTIVE: To define the most effective treatment for severe polyarteritis nodosa (PAN) and Churg-Strauss syndrome (CSS) and to investigate the indication for plasma exchange treatment. METHODS: We conducted a prospective, randomized, multicenter trial in which 62 patients were randomly assigned to receive either prednisone plus cyclophosphamide (intravenous bolus) (group A; n = 28) or prednisone plus cyclophosphamide (intravenous bolus) plus plasma exchanges (group B; n = 34) as first-line treatment for severe PAN or CSS. Factors predicting poor prognosis were renal symptoms, gastrointestinal tract involvement, cardiomyopathy, central nervous system involvement, weight loss > 10% of body weight, and age > 50 years old. Patients with hepatitis B virus-related PAN were not included in this study. The end point of the study was control of the disease (recovery or remission) or death. RESULTS: Clinical symptoms and laboratory findings did not differ significantly in the 2 groups. Initial control of the disease was similar in both groups. Relapse after initial control of the disease was observed in 7 patients (4 in group A and 3 in group B). The mean +/- SD followup period was 31.1 +/- 20 months for group A and 35.9 +/- 16.8 months for group B. At 5 years of followup, 38 patients (61.3%) were cured (16 in group A and 22 in group B), and 5 (8.1%) were in remission without treatment but had not yet completed the cure-defining period of 18 months (3 in group A and 2 in group B). Eight (12.9%) (2 in group A and 2 in group B) were considered to be in clinical remission and required a maintenance regimen of low-dose corticosteroids. Eleven patients died during the study period (7 in group A [25%], 4 in group B [11.8%]). Uncontrolled vasculitis was responsible for 4 deaths (2 in each group), and treatment side effects caused the death of 1 patient in group A. There was no significant difference between the 5-year cumulative survival rates of the 2 groups (75% and 88%, respectively). CONCLUSION: Based on our data, combined treatment with prednisone, cyclophosphamide, and plasma exchanges is not superior to treatment with prednisone and cyclophosphamide alone, and plasma exchanges should not be systematically proposed for initial treatment of severe PAN or CSS. PMID- 7488286 TI - Treatment of calcinosis with diltiazem. AB - OBJECTIVE: To test the hypothesis that the calcium antagonist diltiazem is effective in the treatment of calcinosis. METHODS: Diltiazem, 240-480 mg/day, was given to 4 patients with idiopathic or CREST-related (calcinosis, Raynaud's phenomenon, esophageal dysmotility, sclerodactyly, telangiectasias) calcinosis for 1-12 years. Serial radiographs of the affected areas, using identical technique, and clinical evaluations were obtained. A fifth patient, who did not tolerate diltiazem, received verapamil, 120 mg/day for 18 months. RESULTS: All patients taking diltiazem had a reduction or disappearance of the calcific lesions, with striking clinical improvement. One patient's case was followed for 12 years. The response to diltiazem during the first 5 years of treatment has been previously reported in detail; however, over 7 years of additional treatment, there was further reduction of the lesions. One patient developed a large calcific lesion while receiving verapamil for hypertension, and after verapamil was replaced with diltiazem, there was a dramatic response. Verapamil was ineffective in the fifth patient, who did not tolerate diltiazem. CONCLUSION: Long-term treatment with diltiazem, but not verapamil, is effective in calcinosis. PMID- 7488287 TI - Selection of a gene for apolipoprotein A1 using autoantibodies from a patient with systemic lupus erythematosus. AB - OBJECTIVE: To investigate immunoreactivity of systemic lupus erythematosus (SLE) sera with apolipoprotein A1, (Apo A1), the major lipid-binding protein of high density lipoprotein (HDL). METHODS: Since early attempts to identify Apo A1 autoantibodies using standard enzyme-linked immunosorbent assay (ELISA) and immunoblotting techniques had been unsuccessful, a mouse complementary DNA lambda phage expression library was screened. RESULTS: A selected clone (MA1) was found to have 82% DNA sequence homology to a segment of human Apo A1. Since there were nonconservative substitutions in the MA1 protein and lack of a complete sequence, it was possible that the SLE patient's antibodies were binding MA1 epitopes that were shared by the complete human protein but had not been conformationally accessible using the earlier techniques. Thus, gamma-irradiated ELISA plates were used as an alternative antigen-binding surface for intact human Apo A1, and high titer anti-human Apo A1 autoantibodies were then identified in the sera of 5 more SLE patients. CONCLUSION: These findings show that Apo A1 is immunogenic. Apo A1 antibodies may play a role in the decreased HDL levels and Apo A1:Apo B ratios previously reported to occur in subgroups of SLE patients. PMID- 7488290 TI - In vivo effects of stromelysin on the composition and physical properties of rabbit articular cartilage in the presence and absence of a synthetic inhibitor. AB - OBJECTIVE: To characterize the effects of intraarticular injection of recombinant human stromelysin (SLN) on the matrix composition and physical properties of cartilage from lapine stifle joints and the modulation of these effects by the systemic administration of an N-carboxyalkyl synthetic matrix metalloproteinase inhibitor, L-696,418. METHODS: Female 6-8-week-old New Zealand white rabbits received an intraarticular injection of 100 micrograms activated SLN in 1 stifle joint and buffer in the contralateral control knee; these animals were killed after 1 hour. A separate group of animals received an intravenous injection of either 30 mg/kg L-696,418 or buffer prior to intraarticular injection of SLN. Joints were dissected and analyzed for proteoglycan (PG) loss into joint fluid, tissue biochemical composition, and histology by toluidine blue or anti-VDIPEN antibody staining, or were frozen for physical property analysis. Disks of femoropatellar groove cartilage were harvested from the stifle joint and tested in uniaxially confined compression for determination of electromechanical and mechanical properties. RESULTS: Lapine stifle joints that received injection of SLN without systemic administration of L-696,418 showed a 13-fold increase in loss of PG into synovial fluid. Cartilage from these joints showed significant decreases in streaming potential at 1 Hz and electrokinetic coupling coefficient, but no change in equilibrium modulus, dynamic stiffness, or hydraulic permeability. Systemic treatment with L-696,418 resulted in a significant decrease in loss of PG into joint fluid and elimination of changes in cartilage high-frequency streaming potential and coupling coefficient in joints that were injected with SLN. CONCLUSION: The 1-hour exposure to SLN in vivo resulted in loss of PG and exposure of the VDIPEN epitope of the aggrecan core protein in the superficial region of the tissue near the articular surface. This highly localized degradation resulted in electromechanical behavior changes, but little or no change occurred in mechanical properties. Systemic administration of L 696,418 significantly decreased loss of PG from cartilage and prevented the highly localized tissue degradation and the resultant changes in electromechanical behavior caused by intraarticular SLN injection. PMID- 7488288 TI - Antibody reactivity to the HRES-1 endogenous retroviral element identifies a subset of patients with systemic lupus erythematosus and overlap syndromes. Correlation with antinuclear antibodies and HLA class II alleles. AB - OBJECTIVE: To evaluate the correlation between the presence of antibodies to an endogenous retroviral element-encoded nuclear protein autoantigen, HRES-1, and the presence of other antinuclear antibodies and HLA class II alleles in patients with systemic lupus erythematosus (SLE) and overlap syndromes. METHODS: Antibody reactivities to native and recombinant proteins and synthetic peptides were assessed by counterimmunoelectrophoresis, enzyme-linked immunosorbent assay, and Western blotting. HLA class II alleles were determined by oligonucleotide typing. RESULTS: Forty-eight percent of the 153 patients with autoimmune disease, and 52% of the subgroup with SLE, had HRES-1 antibodies. In contrast, 3.6% of 111 normal donors, and none of 42 patients with the acquired immunodeficiency syndrome or 50 asymptomatic human immunodeficiency virus 1-infected patients, had HRES-1 antibodies. Chi-square analyses revealed a significant association between anti HRES-1 and anti-RNP and an inverse correlation between HRES-1 and Ro/La autoantibodies in patients with SLE or overlap syndromes. Antigenic epitopes of HRES-1 and the retroviral gag-related region of the 70-kd protein component of U1 small nuclear RNP, which share 3 consecutive highly charged amino acids (Arg-Arg Glu), an additional Arg, and functionally similar Arg/Lys residues, represent cross-reactive epitopes between the two proteins. Selective removal of HRES-1 antibodies from sera of HRES-1-seropositive/RNP-seropositive patients by absorption on recombinant HRES-1/glutathione-S-transferase-conjugated agarose beads had no effect on anti-RNP reactivities. A comparative multivariate analysis of HLA class II genes revealed a differential segregation of DQB1 alleles in HRES 1-seropositive versus HRES-1-seronegative patients (P = 0.04). While a relative increase of DQB1*0402 among HRES-1-seropositive patients was noted across ethnic groups (P = 0.02), a decrease of DQB1*0201 and DQB1*0301 was found in white HRES 1-seropositive patients (P = 0.04). CONCLUSION: Autoantibodies to HRES-1 are detectable in a distinct subset of patients with autoimmune disease, primarily in those who do not have antibodies to Ro and La. Anti-HRES-1 and anti-RNP reactivities are mediated by cross-reactive but separate antibody molecules. HLA DQB genes, rather than HLA-DRB or DQA genes, may have a more significant influence on generation of these antinuclear autoantibodies. PMID- 7488289 TI - Association of HLA-B39 with HLA-B27-negative ankylosing spondylitis and pauciarticular juvenile rheumatoid arthritis in Japanese patients. Evidence for a role of the peptide-anchoring B pocket. AB - OBJECTIVE: To investigate the characteristics of HLA-B27 that render susceptibility to seronegative spondylarthropathies. METHODS: Serologic HLA class I typing of Japanese patients with ankylosing spondylitis (AS), juvenile rheumatoid arthritis (JRA), and healthy controls, was performed. HLA-B39 subtypes were determined by polymerase chain reaction-sequence-specific oligohybridization. RESULTS: HLA-B27 was present in 40 of 48 patients with AS (83%), and in only 1 of 210 healthy controls (0.5%). Three of 8 patients (37.5%) who were negative for HLA-B27 were positive for HLA-B39, which was significantly higher compared with the HLA-B27-negative controls (6.2%; P = 0.01). Significant association with HLA-B39 was also noted in the JRA patients (16.7%; P < 0.01), especially in those patients with pauciarticular-onset disease (33.3%; P < 0.01). Ten of 13 HLA-B39-positive patients had subtype B*3901 and 3 had B*3902. CONCLUSION: Because HLA-B27 and HLA-B39 share Glu at position 45 and Cys at position 67, both of which constitute components of the peptide-anchoring B pocket, and because they possess similar peptide-ligand motifs, our results may support either the role of the peptides presented by class I antigens or the importance of Cys at position 67, in the development of spondylarthropathies and pauciarticular-onset JRA. PMID- 7488291 TI - Somatostatin-induced modulation of inflammation in experimental arthritis. AB - OBJECTIVE: To study the antiinflammatory effect of different doses of intraarticular somatostatin in experimental arthritis in rabbits. METHODS: Chronic arthritis was induced by a single injection of fibrin into the knee joint of rabbits previously sensitized to this antigen. The effects of sequential intraarticular injections of somatostatin into the rabbit knee, at doses of 500, 750, and 1,000 micrograms, were monitored by measuring knee joint circumferences and hematologic parameters. The measurements were compared with those obtained following use of triamcinolone acetonide and placebo. At the end of the experiments, the knee joints were examined histologically. RESULTS: Somatostatin treatment induced a statistically significant and dose-related reduction of knee joint swelling. This effect was shorter than that produced by triamcinolone acetonide; however, the antiinflammatory activity elicited by successive doses of triamcinolone acetonide declined both in extent and duration, while the effects of somatostatin remained unchanged at each successive treatment. Histopathologic observations showed that both somatostatin and triamcinolone acetonide reduced the inflammatory signs in the joint structures, although triamcinolone acetonide appeared to be more effective. CONCLUSION: These findings suggest that somatostatin exerts an antiinflammatory effect in this model of experimental arthritis and may represent a valid and safer alternative to corticosteroids for intraarticular therapy of arthritis. PMID- 7488292 TI - Rheumatoid synovial fibroblast adhesion to human articular cartilage. Enhancement by neutrophil proteases. AB - OBJECTIVE: To determine if preexposure of human articular cartilage to activated neutrophils alters rheumatoid synovial fibroblast adhesion to human articular cartilage. METHODS: Human articular cartilage was exposed to either activated neutrophils, interleukin-1, or supernatants obtained from activated neutrophils that had been treated with different protease inhibitors. Radiolabeled rheumatoid synovial fibroblasts were then incubated with the cartilage and the number of counts associated with the cartilage was determined. RESULTS: Pretreatment of human articular cartilage with either activated neutrophils or supernatants obtained from activated neutrophils enhanced subsequent rheumatoid synovial fibroblast adhesion. In contrast, interleukin-1 treatment of cartilage did not alter the adhesion of synovial fibroblasts. The enhanced adhesion could be attenuated by pretreatment of the neutrophil supernatants with either diisopropylfluorophosphonate or EGTA and almost completely abolished by using both inhibitors. CONCLUSION: This study demonstrates that adhesion of rheumatoid synovial fibroblasts to human articular cartilage can be enhanced by exposing the cartilage to proteases released by neutrophils. These results suggest that neutrophil products may play a role in enhancing adhesion of rheumatoid synovium to cartilage in vivo. PMID- 7488293 TI - A reexamination of the relationship between active rheumatoid arthritis and the acquired immunodeficiency syndrome. AB - Three patients with rheumatoid arthritis (RA) that remitted with the development of the human immunodeficiency virus (HIV) infection have been described in the literature, and this has contributed to the belief that RA and HIV infection or the acquired immunodeficiency syndrome (AIDS) cannot coexist. However, a computerized MEDLINE literature search revealed reports of 4 patients who did have active RA and AIDS or HIV infection, as well as other case reports of symmetric polyarthritis compatible with RA in patients with HIV infection. Each of the patients whose RA remitted had received standard disease-modifying antirheumatic drug therapy, and 1 of the 3 had a normal T helper:T suppressor ratio at the time of remission. Of the 4 previously reported patients with active RA and AIDS or HIV infection, all had decreased numbers of T helper lymphocytes. The present report describes a fifth patient with both RA and AIDS and reviews the data concerning the coexistence of these 2 diseases. It appears that active RA may indeed coexist with AIDS. It remains to be seen under what settings HIV may have a disease-modifying effect in RA. These issues have important implications regarding the pathogenesis and therapy of RA, especially in terms of the role of CD4+ lymphocytes and anti-CD4 monoclonal antibody therapy. PMID- 7488294 TI - Carpal tunnel syndrome due to T cell lymphoma. AB - In 2 patients, carpal tunnel syndrome was one of the presenting manifestations of a noncutaneous T cell lymphoma. Infiltration of the carpal tunnel by neoplastic T cells was proven by biopsy in both patients. In 1 case, the carpal tunnel syndrome was associated with eosinophilic fasciitis. These observations emphasize the importance of histologic examination of annular ligaments removed during surgical decompression procedures. PMID- 7488295 TI - Brain abscess mimicking giant cell (temporal) arteritis. PMID- 7488296 TI - Clinical images: Ear ulceration in discoid lupus erythematosus. PMID- 7488297 TI - IgM deficiency in systemic lupus erythematosus patients. PMID- 7488298 TI - Frequency of mannose-binding protein deficiency in patients with systemic lupus erythematosus. PMID- 7488299 TI - Inhibition of leukotriene C4 and B4 release by human eosinophils with the new 5 lipoxygenase inhibitor 6-hydroxy-2(4-sulfamoylbenzylamino)-4,5,7 trimethylbenzothiazo le hydrochloride. AB - Eosinophils generate and release leukotrienes C4 and B4 (LTC4, LTB4) and platelet activating factor (PAF), all of which have the capacity to cause inflammation and tissue injury in the airways. This study has examined the effects of a new 5 lipoxygenase inhibitor, 6-hydroxy-2-(4-sulfamoylbenzylamino)-4,5,7 trimethylbenzothiazo le hydrochloride (CAS 120164-49-0, E6080) on the release of LTC4, LTB4 and PAF by human eosinophils, Eosinophils stimulated by 1 mumol/l calcium ionophore A23187 for 15 min released 37.5 +/- 2.2 ng, 2.3 +/- 0.3 ng and 4.0 +/- 0.3 pmol per 10(6) cells of immunoreactive LTC4, LTB4 and PAF, respectively (mean +/- SEM, n = 4). LTC4 and LTB4 releases were inhibited dose dependently by the addition of E6080 to the cell suspension. The IC50 values were 0.26 mumol/l for LTC4 and 0.23 mumol/l for LTB4. PAF release was not inhibited. These results suggest that E6080 is a potent inhibitor of LTC4 and LTB4 release from eosinophils and may provide a protective effect against bronchoconstriction during late-phase asthmatic responses. PMID- 7488300 TI - Effect of (Z)-11-[3-(dimethylamino) propylidene]-6,11-dihydrodibenz[b,e]oxepin-2 acetic acid hydrochloride on experimental allergic conjunctivitis and rhinitis in rats and guinea pigs. AB - The effect of KW-4679 (Z)-11-[3-(dimethylamino) propylidene]-6,11-dihydrodibenz [b,e] oxepin-2-acetic acid hydrochloride, CAS 140462-76-6; proposed INN: olopatadine) on experimental conjunctivitis and rhinitis was studied in comparison with that of ketotifen (CAS 34580-14-8) using guinea pigs and rats, respectively. KW-4679 was effective in inhibiting the antigen- and histamine induced conjunctivitis by both oral and topical administrations. These effects of KW-4679 were somewhat more potent than those of ketotifen. KW-4679 as well as ketotifen was more effective in inhibiting the histamine-induced conjunctivitis than that seen in antigen-induced conjunctivitis when they were given topically. KW-4679 inhibited the increased dye leakage into the nasal cavity induced not only by antigen in actively sensitized rats but also by histamine perfusion in non-sensitized rats by oral and topical administrations. Similar to ketotifen, the effect of KW-4679 on histamine-induced increase in dye leakage was almost same as that induced by antigen. The potency of KW-4679 was higher than that of ketotifen in the increased dye leakage induced by both antigen and histamine perfusion. PMID- 7488301 TI - Relative bioavailability of carbinoxamine and phenylephrine from a retard capsule after single and repeated dose administration in healthy subjects. AB - The plasma pharmacokinetics of carbinoxamine (CA, CAS 486-16-8) and phenylephrine (PE, CAS 59-42-7) after single dose administration of a retard capsule (Rhinopront) containing 20 mg PE hydrochloride and 4 mg CA maleate, were compared to those of the same active principles given as an aqueous solution. The study was performed in 20 healthy subjects according to a standard crossover design with a one-week wash-out. The pharmacokinetic profile of the active ingredients of the retard capsule was also investigated in the same subjects under repeated dosing conditions (one capsule b.i.d. during 4 days). Blood samples were collected before each administration and up to 36 h after the first and last doses. CA and total PE (free + conjugated) were assayed in the plasma samples by HPLC with coulometric detection and by gas chromatography with electron-capture detection, respectively. The pharmacokinetic parameters obtained after single dose administration indicated an effective slow release of PE and CA with the retard capsule, compared to the solution. Significantly dampened Cmax, delayed tmax and prolonged plateau time were observed. Despite the clear decrease in absorption rate, the two formulations yielded a similar extent of absorption for CA (90% confidence interval of AUC ratio: 61-111%), but not for PE (90% confidence interval of AUC ratio: 56-69%). At steady-state, accumulation of the two active principles apparently followed simple superposition (accumulation index R = 1.6 for PE and 3.9 for CA). The slow absorption pattern of the formulation was maintained at steady-state with tmax and plateau time similar to single dose conditions. PMID- 7488302 TI - Experimental studies on the influence of surfactants on intestinal absorption of drugs. Cefadroxil as model drug and sodium lauryl sulfate as model surfactant: studies in rat duodenum. AB - The effect of sodium lauryl sulfate (CAS 151-21-3) on the duodenal absorption of cefadroxil (CAS 50370-12-2) has been investigated with the aid of a classical rat gut in situ preparation. Both compounds were entirely compatible in working solutions. Cefadroxil was found to be very stable and only slightly solubilized in the micellar phase. The apparent first-order absorption rate constants for the free antibiotic fraction were determined in free solution, and in the presence of variable surfactant concentration in luminal fluid. A functional interpretation of these data, based both on the law of mass action and the complete noncompetitive transport inhibition equations, showed that the surfactant acts as a nonspecific inhibitor of the carrier-mediated absorption of the antibiotic, but also as an enhancer of its passive absorption component. The net result was an outstanding reduction in the absorption capacity of cefadroxil when it was perfused at 0.1 mg/ml, i.e. far from its carrier saturation (from 3.0 h-1 in free solution to 2.0(-1) at high surfactant concentration, with a minimum of about 1.4 h-1 in the presence of the surfactant at 0.5 mg/mg in duodenal fluid). When cefadroxil was perfused at 10.0 mg/ml, i.e. with its carrier-mediated transport beyond the saturation, the net result was a progressively enhanced absorption (ranging from about 0.9 h-1 in free solution to 2.0 h-1 at high surfactant concentration).(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7488303 TI - [In vitro activity of Mercurius cyanatus complex against relevant pathogenic bacterial isolates]. AB - The antimicrobial activity of mercurius cyanatus complex (Oligoplex) and its components Mercurius cyanatus D5, Echinacea angustifolia D1, Ailanthus glandulosa D3, Ammonium bromatum D3, Baptisia tinctoria D3, Euspongia officinalis D2, alcohol 5% (dilution: D1 = 1: 10, D2 = 1 : 100 etc.) was tested in vitro by serial dilution tests against 105 clinical isolates (grampositive/negative, aerobes and anaerobes with relevance for pharyngitis). The bactericidal activity was compared with that of vancomycin when appropriate. One component of the composition (Mercurius cyanatus) exerted a considerable bactericidal activity against S. pyogenes, S. agalactiae, S. pneumoniae, S. aureus, E. faecalis in serial dilutions of the clinical relevant concentration D5. However, growth of H. influenzae, Bacteriodes sp. and Actinobacillus actinomycetemcomitans was not inhibited by Mercurius cyanatus and any other component of the composition. The composition, however, exerted a bactericidal range similar to that of Mercurius cyanatus, but less efficient. Analysis of the bactericidal effect of Mercurius cyanatus and vancomycin revealed comparability for S. pyogenes, S. agalactiae, S. pneumoniae, S. aureus and E. faecalis for vancomycin concentrations of 0.063-2 mg/l, which are clinically relevant. PMID- 7488304 TI - Effect of quaternary ammonium salts on energy-yielding and energy-requiring processes in Salmonella typhimurium. AB - The effect of 2-(dodecanoylamino) ethylalkyldimethyl-ammonium bromides on energy yielding (respiration) and energy-requiring (biosynthesis of nucleic acids and proteins) processes in Salmonella typhimurium cells was studied. The quaternary ammonium salts represented a homologous series (n = 7). The efficacy of the compounds increased with the prolongation of the alkyl carbon chain up to octyl (C8), where the maximum was noted and then the efficacy decreased. This phenomenon known from the antimicrobial efficacy of these substances (cut-off effect) was shown also by influencing the rate of [14C]adenine and [14C]leucine incorporation and the inhibition of oxygen consumption (endogenous respiration). The tested compounds influencing these processes interfere with the energy metabolism of Salmonella typhimurium cells. PMID- 7488305 TI - Inhibitory effects of mixed disulfides from disulfiram on the metabolism and genotoxicity of N-nitrosodiethylamine. AB - Disulfiram (CAS 97-77-8, DSF), a potent anticarcinogenic compound, is known to form mixed disulfides with sulfhydryl group containing amino acids or proteins in vivo. In the present study the stabilities of two mixed disulfides which may arise in the metabolism of disulfiram, i.e. S-(N,N-diethyldithiocarbamoyl)-N acetyl-L-cysteine (AC-DDTC) and S-(N,N-diethyldithiocarbamoyl)-L-glutathione (GS DDTC) in phosphate buffer (pH 7.2) and in rat liver subcellular fractions were investigated as well as their influences on the glutathione (GSH)-related detoxifying system, on the metabolism of [14C] N-nitrosodiethylamine (NDEA) and on the genotoxic activity of NDEA in rats. Both substances were stable in buffer and in microsomes but were degraded in cytosol showing a half life of 4.7 h (AC DDTC) and 3.2 h (GS-DDTC). Addition of GSH to the incubation media accelerated the degradation of mixed disulfides in cytosol. In vivo administration of AC-DDTC and GS-DDTC (1.7 mmol/kg i.p.) led to an increase in hepatic GSH content and to an inhibition of the activity of NDEA deethylase. Both mixed disulfides inhibited the metabolism of NDEA. After a 28 mg/kg i.p. dose of [14C] NDEA only 0.4% was excreted unchanged in the urine. Pretreatment with AC-DDTC and GS-DDTC caused a 10 to 20 fold increase in the amount of NDEA excreted in the urine. The occurrence of DNA single strand breaks in rat liver cells induced by NDEA was completely neutralized by the pretreatment with AC-DDTC. PMID- 7488307 TI - [The effect of noradrenaline and dopamine agonists and antagonists on oxygen consumption in some insect and crustacean species]. AB - The effect of drugs on oxygen consumption in some invertebrate species (insect larvae and crustacea) was investigated. Insect larvae showed an increase in oxygen consumption under the influence of sympathomimetic drugs such as isoprenaline or orciprenaline which could be prevented by beta-receptor antagonists like propranolol or butoxamine. The drugs had no effect in crustacea. On the other hand, in these animals dopamine agonists of different chemical structure (amphetamine, apomorphine, lisuride) caused an increase in oxygen consumption whereas the dopamine antagonist metoclopramide had an opposite effect. PMID- 7488306 TI - Tissue distribution and deposition of clofazimine in rat following subchronic treatment with or without rifampicin. AB - Tissue distribution and deposition characteristics of clofazimine (CAS 2030-63 9), an antileprotic drug in rats have been investigated following controlled sub chronic administration (p.o.) for a period of 1-2 months. The drug was administered alone at a dose of 20 mg/kg body weight and in combination with rifampicin (CAS 13292-46-1) (20 mg/kg p.o.). Various tissues (liver, lung, spleen, small intestine, brain, heart, kidney, skin, stomach and subcutaneous fat) were analyzed for clofazimine in all the treated groups. High levels (range 0.9-3.6 mg/g of wet tissue) were observed in tissues having reticuloendothelial components. In other tissues the levels were relatively lower (range 3-114 micrograms/g of wet tissue). Histopathological studies revealed that clofazimine is deposited in many tissues in the form of reddish-orange crystals. Concomitant treatment with rifampicin did not significantly alter tissue distribution or deposition profile of clofazimine nor did it influence the histopathology. PMID- 7488308 TI - [New horizons for medicine on the basis of genetic technology]. AB - In case of a single gene defect a number of appropriate gene probes are available for prenatal diagnosis. Knowledge of the genetic disorders enables in some cases early onset of therapy or the option for abortion. However, gene technology which enables the diagnosis should not be viewed from an ethical point of view but rather the action taken when diagnostic results are available. Gene therapy for a single gene defect still is at the early stage of development. Only few patients have been treated in various indications. Difficult to be overcome are the low frequency and unspecific integration of inserted DNA into the chromosome, lack of sufficient transcription control and short half-lives of the integrated gene. From an ethical perspective gene therapy complies with the therapeutic concept of medicine. Antisense oligonucleotides are under clinical development for blockage of the synthesis of oncogenes and viral proteins. Stability of obligonucleotides as well as selectivity for specific cells will have to be overcome for broader application. Its therapeutic application is in accordance with the ethical principles of medicine. Substitution therapy with recombinant DNA derived human proteins is in therapeutic application to replace their counterparts from native source by human pharmacologically active proteins which cannot be isolated from their natural source. For recombinant DNA derived proteins where the mode of action is known short development times frames can be expected allowing an early return on investment. The expected market potential for recombinant DNA derived pharmaceuticals in 1995 will reach 4.4 billion DM. PMID- 7488309 TI - Screening and identification of sumatriptan and its main metabolite by means of thin-layer chromatography, ultraviolet spectroscopy and gas chromatography/mass spectrometry. AB - Sumatriptan (CAS 103628-46-2, 3-[2-(dimethylamino)ethyl]-N-methyl-1H-indole-5 methanesulphonamide++ +), a substance for treatment of acute migraine attacks, and its main metabolite are investigated by thinlayer chromatography (TLC), ultraviolet spectroscopy, and gas chromatography/mass spectrometry (GC/MS). The resulting analytical data (correlated hRf-values, UV solvent spectra, remission spectra, GC retention indices, and electron impact (EI) mass spectra) including an extraction procedure and different derivatization methods are presented. Their applicability is described for urine analysis. PMID- 7488310 TI - Synthesis of new serotonergic 2-substituted 4,6-diaryl pyridazin-3-ones. AB - A series of 4,6-diaryl pyridazin-3-ones substituted in the 2-position by [4-(4 aryl piperazin-1-yl]-but-2-ynyl moieties was synthesized and evaluated for antidepressant activity. The structures of these new pyridazine derivatives were confirmed by IR, 1H-NMR spectra and by elementary analysis. At 150 mg/kg i.p., they induced little or no reduction of the duration of immobility of mice in the forced swimming test. Head twitches produced by L-5-hydroxytryptophan in mice pretreated with pargyline were significantly potentiated by most of the tested compounds. In addition, pyridazine derivatives did not antagonize reserpine induced palpebral ptosis or enhance the toxic effects of yohimbine and were almost devoid of anticholinergic properties in mice. PMID- 7488311 TI - Cerebral blood flow effects of sumatriptan in drug combinations in the baboon model. AB - Sumatriptan (CAS 103628-46-2, Imigran) has established itself as an important therapeutic agent in the treatment of migraine. Although considerable understanding of, in particular, the vascular pathophysiology of migraine has been gained during the past decade, the pathophysiology and mediators involved in the pain experience during migraine ar during migraine are not yet fully explained. The mechanisms behind the pharmacological effects of sumatriptan are still only partially understood. In the present study the effects of sumatriptan on drug induced cerebral blood flow increases in the baboon model were investigated using 99mTc-HMPAO (hexamethylpropylene amine oxime) and SPECT (single photon emission computed tomography). Sumatriptan selectively reduced drug induced cerebral blood flow increases. The effects of halothane anaesthesia and acetazolamide on cerebral blood flow were not reversed by sumatriptan, while the effect of nimodipine was attenuated by 47% (to the level of cerebral blood flow below the normal flow baseline). These results support multiple mechanisms for sumatriptan involving vascular neurotransmission and neurogenic inflammatory responses via serotonin receptor stimulation and Ca2+ mobilization. Drug-drug interactions are further implicated through this study. PMID- 7488312 TI - Structure-activity relationship studies in the field of calcium antagonists. Xanthone 1,4-dihydropyridines bearing a 2,3-lactone ring. AB - A series of xanthone 1,4-dihydropyridine derivatives bearing a 2,3-lactone ring and a 2-acetoxymethyl group were prepared. The compounds were evaluated for inotropic, chronotropic and calcium antagonist properties. The introduction of a 2,3-lactone ring improved the negative inotropic activity and selectivity. PMID- 7488313 TI - Tolerance development and arterial and venous tissue-/plasma levels of glyceryl trinitrate and glyceryl dinitrate in patients treated with nitroglycerin infusion. AB - Venous vessels (vena saphena magna) and arteries (left internal mammary artery) from patients treated with long-term ( > 24 h) intravenous infusions of nitroglycerin (CAS 55-63-0, glyceryl trinitrate, GTN) prior to coronary by-pass surgery were studied. Tissue concentrations of GTN and its dinitrate metabolites were determined as well as plasma-concentrations of GTN and glyceryl dinitrates (GDN's) on both the arterial and the venous side. The relaxant effect of GTN on the isolated vein preparations was studied and compared with those from two other groups of patients, one not exposed to GTN and the other exposed to a short-term infusion of GTN. Wide variations were found both in plasma and in tissue concentrations of GTN and GDN's on the venous and the arterial side. However, no extraction across the arterial-venous bed could be observed. Correlations were found between the dose given and the concentrations of drug and metabolites in both venous and arterial plasma as well as between 1,2-GDN in venous plasma and venous tissue, between 1,2-GDN in arterial plasma and arterial tissue and the same applied to 1,3-GDN on the arterial side. No correlations were found between the tissue concentrations of drug or metabolites and the relaxation induced by GTN in vitro. Venous vessels from patients treated with long-term infusions of GTN showed a pronounced decrease in relaxation as compared to vessels from patients not treated with nitroglycerin infusions and a slight decrease as compared to veins from patients treated with a shortterm infusion. PMID- 7488314 TI - Binding affinities of isocarbacyclin methyl ester and its free acid to prostanoid receptors. AB - The binding affinities of isocarbacyclin methyl ester (CAS 88931-51-5, TEI-9090) and its free acid (CAS 88911-35-7, TEI-7165) to prostaglandin (PG) I2 receptors (IP receptors), PGE2 receptors (EP receptors) and thromboxane A2 receptors (TP receptors) were investigated. TEI-9090 exhibited low affinity for IP and EP receptors in mastocytoma P-815 cell membranes and no affinity for TP receptors in washed guinea-pig platelets. The IC50 values of TEI-9090 against [3H]iloprost (for IP receptors), [3H]PGE2 (for EP receptors) and [3H]SQ29,548 (for TP receptors) binding were 2803 +/- 327 nmol/l, 2509 +/- 1317 nmol/l and > 10000 nmol/l (n = 3), respectively. In contrast, TEI-7165 had high affinity for IP receptors (IC50 = 65.4 +/- 28.5 nmol/l, n = 3) but had moderate to low affinity for EP and TP receptors. The affinity of TEI-9090 for IP receptors was remarkably enhanced by pretreatment with rat serum as an esterase source. These findings suggest that TEI-9090 is converted to TEI-7165 by esterase, and that TEI-7165 exerts pharmacological effects through binding to IP receptors. PMID- 7488315 TI - Exhibition of anti-platelet effect of isocarbacyclin methyl ester and its free acid via an increase in cAMP in platelets. AB - Isocarbacyclin methyl ester (CAS 88931-51-5, TEI-9090) and its free acid (CAS 88911-35-7, TEI-7165) are chemically stable analogues of prostaglandin (PG) I2. The cAMP increasing activities of TEI-9090 and TEI-7165 were investigated. TEI 9090, TEI-7165, PGI2, and PGE1 caused the accumulation of cAMP in rabbit and human platelets. The effects of TEI-9090, TEI-7165, and PGE1 were longer lasting than the effect of PGI2. Each drug concentration-dependently increased the cAMP level in rabbit and human platelets. The order of cAMP-increasing activity was PGI2 > PGE1 = TEI-9090 = TEI-7165 in rabbit platelets and PGI2 > TEI-9090 = TEI 7165 > PGE1 in human platelets. The antiplatelet aggregation activity of TEI-9090 in rabbit and human platelets was remarkably decreased by the addition of an esterase inhibitor, diisopropyl fluorophosphate (DFP). However, this activity of TEI-7165 was not affected by the inhibitor. In the presence of DFP, the cAMP accumulation by TEI-9090 in rabbit platelets was almost completely inhibited but that by TEI-7165 was not inhibited. These results suggest that TEI-9090 is deesterified to TEI-7165 and that this free acid then inhibits platelet aggregation via an increase in the cAMP level in the platelets. PMID- 7488316 TI - Anti-platelet effects of isocarbacyclin methyl ester on human and rabbit platelets in vitro. AB - Anti-platelet effects of isocarbacyclin methyl ester (CAS 88931-51-5, TEI-9090) and of its free acid form, TEI-7165 (CAS 88911-35-7) were compared with those of prostaglandin (PG) I2 and PGE1. TEI-9090 and TEI-7165 dose-dependently inhibited human and rabbit platelet aggregation and human platelet adhesion in vitro. The IC50 values of TEI-9090, TEI-7165, PGI2, and PGE1 against ADP-induced human (rabbit) platelet aggregation were 22.90 (25.00) ng/ml, 2.78 (6.46) ng/ml, 1.67 (3.34) ng/ml, and 19.91 (8.32) ng/ml, respectively; and those against human platelet adhesion were 3.47 ng/ml, 0.13 ng/ml, 0.25 ng/ml, and 1.45 ng/ml, respectively. TTC-909, a product incorporating TEI-9090 in lipid microspheres, had an aggregation inhibitory effect similar to that of TEI-9090 in human platelets. The aggregation inhibitory effect of TEI-9090 and TTC-909 increased with incubation time, and reached a level similar to that of TEI-7165. In contrast, the aggregation inhibitory effect of TEI-7165 remained constant for 120 min, whereas that of PGI2 decreased with time. TEI-9090 and TEI-7165 also enhanced the disaggregation of human platelets at nearly the same concentration as they exerted their inhibitory effect on aggregation. The order of anti platelet activities, compared at each optimum incubation point, were PGI2 approximately equal to TEI-7165 approximately equal to TEI-9090 (approximately equal to TTC-909) > PGE1 in human and rabbit platelets. These results indicate that TEI-9090 (TTC-909) and its active metabolite, TEI-7165, may be a potent anti platelet drug. PMID- 7488317 TI - Ex vivo anti-platelet effects of isocarbacyclin methyl ester incorporated in lipid microspheres in rabbits. AB - Effect of TTC-909, a chemically stable isocarbacyclin methyl ester (CAS 88931-51 5, TEI-9090) incorporated in lipid microspheres, on platelet aggregation was tested in conscious rabbits. TTC-909 significantly inhibited platelet aggregation induced by ADP at early times after bolus i.v. injection from the dose of 0.3 micrograms/kg in a dose-dependent manner. Similarly, TTC-909 significantly inhibited platelet aggregation induced by ADP during the venous infusion from the dose of 100 ng/kg/min, also in a dose-dependent manner. The anti-platelet effect of TTC-909 was observed at a dose lower than that inducing hypotensive effect when given as both bolus i.v. and venous infusion. On the other hand, prostaglandin (PG) E1 (LM)--PGE1 incorporated in lipid microspheres--reduced blood pressure without having an anti-platelet effect. These results indicate that TTC-909 may be a potent anti-thrombotic drug. PMID- 7488318 TI - Vasorelaxant effect of isocarbacyclin methyl ester incorporated into lipid microspheres on isolated canine arteries. AB - TTC-909 is a preparation of isocarbacyclin methyl ester (CAS 88931-51-5, TEI 9090) incorporated into lipid microspheres. The characteristics of vasorelaxation induced by TTC-909 were investigated in isolated canine arteries. TTC-909 produced concentration-dependent relaxation of prostaglandin F2a(PGF2a)-induced and U-46619 (TXA2 receptor agonist)-induced contraction in basilar, coronary, renal, mesenteric, and femoral arteries. The vasorelaxant potency of TTC-909 was compared with the potencies of TEI-9090, TEI-7165 (CAS 88911-35-7, free acid of TEI-9090), prostaglandin I2 (PGI2), and prostaglandin E1 (PGE1) on PGF2a-induced contraction in basilar and femoral arteries. The relaxing potency of TTC-909 was similar to the values for TEI-9090, TEI-7165, and PGI2, and more potent than that of PGE1 in both of these arteries. An esterase inhibitor, diisopropyl fluorophosphate (DFP), reduced the vasorelaxant effect of TEI-9090, but not that of TEI-7165, on PGF2a-induced contraction in femoral arteries. These results suggest that TTC-909 has vasorelaxant potency similar to that of PGI2 and greater than that of PGE1. Further, it appears that TEI-9090 was converted to TEI-7165 by esterase and then produced its vasorelaxing effect. PMID- 7488319 TI - Effect of isocarbacyclin methyl ester incorporated into lipid microspheres on cutaneous blood flow in rat. AB - A new stable prostacyclin analogue, isocarbacyclin methyl ester (CAS 88931-51-5, TEI-9090), incorporated into lipid microspheres (TTC-909), was examined for its effects on cutaneous blood flow and mean blood pressure in rats in comparison with PGE1 (LM) (prostaglandin E1 in lipid microspheres). TTC-909 at 0.3 micrograms/kg i.v. significantly increased the sole cutaneous blood flow without changing blood pressure. A significant decrease in mean blood pressure was observed with TTC-909 above 1 microgram/kg i.v. PGE1 (LM) also showed a decrease in the blood pressure from 1 microgram/kg i.v., whereas the sole cutaneous blood flow was not affected even at 10 micrograms/kg i.v. These results suggest that TTC-909 dilates the skin vessels preferentially to other blood vessels in the rat and that effect is more potent than that of PGE1 (LM). TTC-909 may thus prove to be an effective agent for the treatment of peripheral vascular disease. PMID- 7488320 TI - Effect of isocarbacyclin methyl ester incorporated in lipid microspheres on experimental models of peripheral obstructive disease. AB - The effect of TTC-909 (isocarbacyclin methyl ester (CAS 88931-51-5, TEI-9090) incorporated into lipid microspheres) on obstruction of the peripheral artery was studied in two different animal models. The first was a peripheral occlusion model induced by intra-arterial injection of sodium laurate in rats. The second was a tail gangrene model induced by subcutaneous injection of both ergotamine and epinephrine in rats. Intravenous bolus-administered TTC-909 at 1.0 microgram/kg resulted in the inhibition of both progression of lesions induced by sodium laurate and the extension of tail gangrene by ergotamine and epinephrine. Moreover, the inhibitory effect of TTC-909 was more potent than that of PGE1 (LM) (prostaglandin E1 incorporated into lipid microspheres). These findings suggested that TTC-909 may be clinically useful for the therapy of peripheral vascular disorders such as thromboangiitis obliterans (TAO) and atherosclerosis obliterans (ASO). PMID- 7488321 TI - Two different inhibitory effects of pyridoxalphosphate-6-azophenyl-2',4' disulfonic acid on adenosine diphosphate-induced human platelet aggregation. AB - In the present study, the novel compound pyridoxalphosphate-6-azophenyl-2',4' disulfonic acid (PPADS), which has been shown to inhibit P2x-purinoceptor mediated contractions in smooth muscle, was investigated for its antagonistic effects on adenosine diphosphate (ADP)-induced human platelet aggregation in platelet-rich plasma (PRP) and in washed platelets, respectively. Suramin served as reference compound. In PRP, suramin (1 mmol/l) was inactive whereas PPADS (1 mmol/l) considerably reduced the extent of aggregation. In contrast, both suramin (1 mmol/l) and PPADS (500 mumol/l) markedly depressed the aggregation of washed platelets. In addition, there was a peculiarity in washed platelets: a delay of onset of platelet aggregation up to 15 min in the presence of PPADS (10-500 mumol/l) and suramin (0.1-1 mmol/l). Thus, in the present study washed platelets were more suited to detect an influence of PPADS and suramin as inhibitors of ADP induced aggregation, and a delay of onset of aggregation was the most sensitive parameter in this regard. Comparing the effective threshold concentration of PPADS at P2x (1 mumol/l)- and the platelet P2t-purinoceptor, PPADS proves to be P2x-selective. PMID- 7488322 TI - Synthesis of 1,2-diacyl-3-nicotinoyl glycerol derivatives and evaluation of their acute effects on plasma lipids in the rat. AB - Nicotinic acid (CAS 59-67-6) is the only hypolipidemic agent whose activity has been shown both on atherosclerotic lesions and on long term mortality. Unfortunately, its use is hindered by the frequent occurrence ( > 70%) of adverse reactions (i.e. cutaneous rash, pruritus and, most significantly, flush). New prodrugs of nicotinic acid have been prepared by the use of diacylglycerol esters. In the rat, after acute oral administration of these products, a significant decrease of the free fatty acid plasma levels was obtained without the dramatic increase in nicotinic acid plasma levels observed after the oral administration of an equimolecular dose of nicotinic acid. The most interesting ester, S 16961 ((d,l)-1,2-dipalmitoyl-3-nicotinoyl glycerol, CAS 160555-46-4) is undergoing clinical trials. PMID- 7488323 TI - ASHA's 1995 annual convention. Orlando, Florida, December 7-10, 1995. Abstracts. PMID- 7488324 TI - Is reverse cholesterol transport a misnomer for suggesting its role in the prevention of atheroma formation? AB - Reverse cholesterol transport from peripheral tissues, including the arterial wall, involves high density lipoprotein (HDL) uptake of unesterified cell cholesterol, its esterification by lecithin-cholesterol-acyl-transferase (LCAT), direct HDL-cholesteryl ester uptake by the liver and the indirect pathway consisting of the cholesteryl ester transfer protein (CETP)-mediated transfer of HDL-cholesteryl ester to apolipoprotein (apo) B-containing lipoproteins (very low density lipoprotein (VLDL) and LDL). Although the first route should be regarded as anti-atherogenic, ambiguous interpretations are drawn from the indirect pathway since it is potentially atherogenic to the extent that it may raise the plasma cholesteryl ester concentration in lipoproteins that are taken up by arterial wall macrophages. In addition, controversial roles are played in reverse cholesterol transport by LCAT and liver uptake of HDL-cholesteryl ester mediated by hepatic lipase (HL). HDL may exert several antiatherogenic effects unrelated to its role in cell cholesterol removal. PMID- 7488325 TI - Deposition pattern of monocytes and fatty streak development in hypercholesterolemic rabbits. AB - To further characterize the role of monocytes in atherogenesis, we studied the influence of a qualitatively, well-defined hemodynamic flow field on the deposition pattern of monocytes in the thoracic aorta of normal (N, n = 6) and hypercholesterolemic (H, n = 10) rabbits. Pairs of H rabbits were sacrificed after 1, 2, 4, 7 and 10 weeks of cholesterol feeding. Complete deposition patterns of adherent cells were quantified over 500 mm2 of aortic endothelium around the lesion-susceptible intercostal orifices using an en face light microscopic technique. Adherent cells were almost exclusively monocytes by morphological criteria and non-specific esterase staining. The mean density of adherent cells in normal rabbits was 1.28 +/- 1.21 (S.D.) per mm2 of endothelium and increased nearly 5-fold by 7 weeks of cholesterol feeding. High local densities of adherent monocytes (up to 34 cells/mm2) were noted over early fatty lesions present in one 4 week and all 7 and 10 week H rabbits. Adherent cell densities near intercostal orifices prior to lesion formation were approximately 50% greater than in non-orifice regions in both the normal and the 1 and 2 week H rabbit groups. These differences were statistically significant at P < 0.05 by ANOVA. We conclude that preferred adherence of monocytes occurs around intercostal orifices in normolipidemic and early cholesterol-fed rabbits before lesions develop at these lesion-prone sites. Monocyte deposition appears to be governed not only by the arterial flow field but also by cholesterol feeding since higher numbers of adherent monocytes were found on both early fatty streaks and nonlesioned endothelium in rabbits fed cholesterol for longer than 4 weeks. PMID- 7488326 TI - Vitamin K intake and osteocalcin levels in women with and without aortic atherosclerosis: a population-based study. AB - Protein-bound gamma-carboxyglutamate (Gla) has been demonstrated in calcified atherosclerotic plaques. Vitamin K is required for the formation of Gla-residues. As the biological activity of Gla-proteins appears to be strictly dependent on the presence of the Gla-residues, vitamin K status may be an important factor in the development and progression of atherosclerotic calcifications. We studied the association of vitamin K status, as assessed by nutritional vitamin K intake and the measurements of two circulating immunoreactive osteocalcin (irOC) fractions, with aortic atherosclerosis in a population-based study of 113 postmenopausal women. Women with calcified lesions (n = 34) had a 42.9 micrograms lower mean age adjusted dietary vitamin K intake/day (95% C.I. -6.6 to 92.5) than those without calcifications (n = 79). Atherosclerotic women had higher irOC levels with a low affinity for hydroxyapatite (irOCfree): age-adjusted difference of 0.32 ng/ml (95% C.I. 0.03 to 0.61). In addition, the high affinity irOC levels expressed as a percentage (hydroxyapatite binding capacity, HBC) were 5.12% (95% C.I. 1.32 to 8.92) lower in women with calcifications. Our study indicates that women with aortic atherosclerosis have an impaired vitamin K status as reflected by a lower nutritional vitamin K intake, an increased irOCfree level and a reduced HBC level. An impaired vitamin K status in subjects with atherosclerosis is compatible with the view that vitamin K or Gla-containing proteins are involved in the development of calcification of the vessel wall. PMID- 7488327 TI - Inhibitory effects of arachidonic acid (20:4,n-6) and its monohydroperoxy- and hydroxy-metabolites on procoagulant activity in endothelial cells. AB - The procoagulant response of endothelium to pathophysiological agents such as tumour necrosis factor alpha (TNF alpha) and phorbol myristate acetate (PMA) alters the expression of proteins such as tissue factor. The modulation of such procoagulant activity (PCA) by the polyunsaturated fatty acid arachidonic acid (20:4,n-6) and its 15-hydroperoxy (15-HPETE) and 15-hydroxy (15-HETE) metabolites was examined since this may have important implications in cardiovascular disease and atherosclerosis. Treatment of human umbilical vein endothelial cells (HUVEC) for 30 min with 20:4, 15-HPETE or 15-HETE before induction of PCA with TNF alpha (100 U) or PMA (10(-7) M) caused a significant inhibition of PCA. This inhibition was seen at 2-5 microM fatty acids. Dose response curves with TNF alpha indicated that the inhibition was greatest at higher concentrations of TNF alpha (> or = 250U TNF alpha/ml). The mode of administration of the fatty acid was not critical as fatty acids presented as DPC-fatty acid micelles or solubilised in ethanol gave similar inhibitions of PCA. 20:4, 15-HPETE or 15-HETE did not alter the binding of I125-labelled TNF alpha to its surface receptors on HUVEC, suggesting that the effect of these fatty acids was not mediated by events at the cell surface receptor level. In support of this, these fatty acids were found to inhibit PCA induced by PMA which bypasses cell surface receptors to activate protein kinase C directly.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7488329 TI - Post-methionine load hyperhomocysteinemia in persons with normal fasting total plasma homocysteine: initial results from the NHLBI Family Heart Study. AB - Hyperhomocysteimia, either fasting or after oral methionine loading, appears to be an independent risk factor for coronary heart disease (CHD). It remains unclear whether fasting total homocysteine determination alone adequately detects the full spectrum of hyperhomocysteinemic individuals. We measured fasting and 4 h post methionine loading (0.1 g L-methionine/kg body weight) total plasma homocysteine in 274 participants in The NHLBI Family Heart Study, a population based investigation of genetic and non-genetic determinants of CHD. Of the total number (n = 47) of hyperhomocysteinemic persons, 43% (20/47) were identified only by methionine loading, while 32% (15/47) of the total number, and 75% of those with post-methionine loading hyperhomocysteinemia only (15/20), had fasting total homocysteine concentrations below the 75th percentile (10.7 mumol/l). We conclude that fasting total plasma homocysteine determination alone fails to identify a sizable percentage (> 40%) of persons who may have clinically relevant hyperhomocysteinemia post methionine loading. PMID- 7488328 TI - Common variation in the gene for apolipoprotein B modulates postprandial lipoprotein metabolism: a hypothesis generating study. AB - We have carried out a pilot study to examine the influence on postprandial lipid and lipoprotein metabolism of common genetic variation in the gene coding for apolipoprotein (apo) B, in a previously described group of 30 individuals who had survived a myocardial infarction (MI) before the age of 45 (normo (NTG)- and hypertriglyceridaemic (HTG) patients) and 11 age-matched healthy individuals. Postprandial lipid or lipoprotein levels were examined by genotypes in the three groups separately and after adjustment for fasting triglycerides (TG) in the whole group combined. For the signal peptide polymorphism in the apo B gene, individuals with one or more SP-24 alleles had a 38% smaller mean area under curve (AUC) (P = 0.06) for postprandial large chylomicron remnants and a 29% smaller mean AUC (P = 0.01) for large very low density lipoproteins (VLDLs) compared to individuals homozygous for the wild type SP-27 allele. Previously in this patient group, small chylomicron remnants (apo B-48 levels in the Sf 20-60 range) were found to relate significantly and positively to progression of coronary atherosclerosis suggesting that these lipoproteins are implicated in progression of atherosclerosis. For the apo B Val591-Ala polymorphism (Ag a1/d), individuals homozygous for the V591 allele had a 33% greater AUC for Sf 20-60 postprandial triglycerides (P = 0.006), with higher postprandial levels of both apo B-48- and apo B-100-containing lipoproteins in this fraction. This pilot study gives insight into the mechanisms of the previously reported associations between polymorphisms in the apo B gene and fasting plasma lipids and lipoproteins. PMID- 7488330 TI - Oxidized low density lipoprotein stimulates collagen production in cultured arterial smooth muscle cells. AB - We examined the interactive effect of oxidized low density lipoprotein (LDL) and ascorbic acid on collagen production in cultured smooth muscle cells (SMCs). Porcine aortic SMCs were incubated with 50-200 micrograms/ml of human LDL with/without 5 microM Cu2+ for 24 h. Collagen production was assayed by successive salt precipitation at acidic and neutral pH after pepsin digestion of 3H-proline-labeled collagenous protein. Oxidation of LDL was evaluated by electrophoresis and by the level of thiobarbituric acid reactive substances (TBARS). Ascorbic acid reduced the oxidation of LDL + Cu2+ (53% reduction). In the presence of ascorbic acid, no differences were noted in collagen production between LDL and LDL + Cu2+. Without ascorbic acid, collagen production with LDL + Cu2+ was increased dose-dependently up to 6-fold with 150 micrograms/ml LDL, while no such effects were observed at any doses of native LDL. The addition of butylated hydroxytoluene to LDL + Cu2+ strongly suppressed oxidation (88% reduction), and significantly reduced collagen production close to that seen with native LDL. These results indicate that oxidized LDL stimulates collagen production in SMCs, while native LDL does not. Therefore, oxidized LDL may play a direct role in stimulating collagen production in SMCs, which could lead to collagenosis in atherosclerosis. PMID- 7488331 TI - Visualization of the transport pathways of low density lipoproteins across the endothelial cells in the branched regions of rat arteries. AB - The transport pathways of low density lipoproteins (LDL) across the endothelium at the branched and unbranched regions of the artery were studied in high cholesterol diet-fed rats. Rat tissues were analyzed by perfusing in situ human or rat LDL labeled with colloidal gold or fluorescein 1,1'-dioctadecyl-3,3,3',3' tetramethylindocarbocyanine perchlorate (DiI). Results indicated that more LDL DiI accumulated in the branched regions than in the unbranched regions of the artery. LDL-gold conjugates were observed in the plasmalemmal vesicles, multivesicular bodies and in the subendothelial space in both the branched and the unbranched regions of the arteries. Quantitative study revealed that the volume densities of plasmalemmal vesicles which contained the LDL-gold particles in the branched regions of the aortic arch were significantly (P < 0.05) higher than the density value in the unbranched regions of the thoracic aorta, whereas there was no marked difference in the density value of multivesicular bodies between these two regions. The open junctions with gap widths of 30-450 nm between adjacent endothelial cells were only observed in the branched regions of the aortic arch, whereas no open junctions were present in the unbranched regions of the thoracic aorta. Moreover, the LDL-gold conjugates were present within most of these open junctions. In all specimens examined, no gold particles were found in the normal intercellular channels (i.e., 25 nm and less) of both regions. These results indicated that the major visible routes for transport of LDL across the endothelium in the branched regions of the arteries are open junctions as well as plasmalemmal vesicles. The region-associated permeability changes of LDL might account for the incidence of atherosclerosis in the branched areas of arteries. PMID- 7488333 TI - Net uptake of plasma homocysteine by the rat kidney in vivo. AB - Hyperhomocysteinemia is a common finding in dialysis-dependent end-stage renal disease (ESRD) patients, but its etiology and refractoriness to standard homocysteine-lowering B-vitamin therapy are poorly understood. In the absence of actual in vivo data, it has been hypothesized that loss of normal renal parenchymal uptake and metabolism of homocysteine is an important determinant of hyperhomocysteinemia in ESRD, given that urinary homocysteine excretion by healthy kidneys is trivial. We assessed net renal uptake and metabolism of homocysteine using an established rat model for measuring arteriovenous amino acid differences across the rat kidney, along with simultaneous determination of renal plasma flow, urine flow, and urinary homocysteine concentration. Substantial homocysteine uptake and metabolism by normal rat kidneys was demonstrated, and we also confirmed that urinary homocysteine excretion is minimal. These data suggest that loss of the sizable homocysteine metabolizing capacity of the intact kidneys may be an important determinant of the refractory, potentially atherothrombotic hyperhomocysteinemia frequently observed in ESRD. PMID- 7488332 TI - Proteinases and restenosis in the human coronary artery: extracellular matrix production exceeds the expression of proteolytic activity. AB - To understand the balance of proteinase antiproteinase activity and the production of extracellular matrix (ECM) at the site of arterial injury, we analyzed the composition of ECM and proteinase activity in normal internal mammary arteries, tissue samples obtained from atherosclerotic coronary lesions and restenotic lesions obtained during directional coronary atherectomy. Histologically and biochemically, collagen and proteoglycans increased, and elastin decreased in samples from restenotic lesions when compared to samples taken from patients undergoing their first revascularization (de novo). In contrast, cellularity was increased in samples obtained from de novo patients as compared to samples obtained from restenotic lesions. Intrinsic activity of matrix metalloproteinases (MMPs) was measured by using zymography and scanning all the lytic bands in zymographic gel. In these gels, identical amounts of total protein were loaded in each lane. MMP activity was determined as % of the total (latent and active) MMPs after trypsin activation (100%) in the normal artery. Intrinsic MMP activity was reduced to 6% +/- 1% in atherosclerotic lesions and 1% +/- 1% in restenotic lesions, when compared to activity found in normal (10% +/- 3%) arteries. Based on solubilization of fluorescein-conjugated elastin by the extracts, the MMP-mediated elastinolytic activity was 0.2 +/- 0.1, 8.8 +/- 1.5, and 24.0 +/- 3 nmol/min/mg in restenotic, native atherosclerotic and normal tissue, respectively. The results suggested that, in arterial tissue from patients with angiographic restenosis, there is an increased production of ECM collagen and a decrease in MMP activity compared to both normal artery and atherosclerotic samples from de novo patients undergoing an initial revascularization procedure of a significant coronary artery lesion. PMID- 7488334 TI - Nonenzymatic glycation and extractability of collagen in human atherosclerotic plaques. AB - The relationship between the extractability of collagen by enzymatic digestion and the degree of nonenzymatic glycation of collagen was examined in the aorta and skin from 38 subjects without diabetes mellitus (mean age: 62.3 +/- 20.2 years). Samples were obtained from the aortic media (M), lesion-free intima (I), atherosclerotic intima (A) and dermis of the skin (S). Collagen was extracted first by incubation with 1/50 (enzyme/substrate weight ratio) pepsin at 4 degrees C for 24 h (P-fraction) and then by incubation with 1/10 (enzyme/substrate weight ratio) pepsin at room temperature for 24 h (EP-fraction). The pepsin-insoluble precipitates were digested by incubation with 270 units of bacterial collagenase at 37 degrees C for 24 h (PIS-fraction). Collagen contents, ketoamines and collagen-linked fluorescence (CLF) were measured in each fraction. The amount of ketoamines and the level of CLF correlated inversely with the susceptibility of collagen to pepsin digestion in various tissues, including M, I, A and S. These values were highest in both the P- and EP-fractions of M, which contained the least amount of collagen extracted by pepsin digestion. In contrast, they were lowest in S, where the concentration of collagen extracted by pepsin digestion was greatest among all of the tissue samples. Atherosclerotic intima (A) and aortic media (M) showed an age-related increase in the total amount of collagen digested with pepsin and collagenase, which depended mainly on an increase in the content of pepsin-insoluble collagen. Although the total amount of collagen did not increase with advancing age in I or S, collagen in I and S became progressingly resistant to pepsin digestion. These results suggest that the age related decrease in the susceptibility of collagen to pepsin digestion may be due to nonenzymatic glycation in atherosclerotic lesions as well as normal tissues, including the aortic media, lesion-free intima and skin. The level of CLF significantly increased with age in the P-fraction and/or EP fraction of M, I and S. However, there was no relationship between the level of CLF and the subject's age in A. Thus, the accumulation of advanced glycation endproducts (AGEs) on collagen fibers may be partially responsible for the increase in collagen matrix in atherosclerotic lesions of subjects without diabetes mellitus. PMID- 7488335 TI - Correlation between the presence of an immature smooth muscle cell population in tunica media and the development of atherosclerotic lesion. A study on different sized rabbit arteries from cholesterol-fed and Watanabe heritable hyperlipemic rabbits. AB - Mapping the distribution of an immature smooth muscle cell (SMC) subpopulation in large- and small-sized arterial vessels was carried out in normocholesterolemic rabbits and compared with the mapping atherosclerotic lesions in endogenously (Watanabe heritable hyperlipemic, WHHL) and exogenously derived (cholesterol-fed, CT) hypercholesterolemic rabbits. This cell subset is identified by a specific myosin isoform content and displays an intermediate degree of differentiation between fetal- and adult-type SMC. Monoclonal anti-myosin antibodies, immunofluorescence procedures, and different arterial segments of a rabbit vessel tree, i.e. from aorta to dental pulp (common carotid, external carotid, lingual, facial, maxillary, inferior alveolar arteries, and dental branches of alveolar arteries) were studied. WHHL of different ages (3 to 12 months), and two different concentrations of CT (2% and 0.2%) in the diet for 3 and 12 months, respectively, were used. The results of the present study indicate that: (1) using a diet with a higher percentage of CT (rabbits fed 2% CT-diet for 3 months) there is maximum expansion of atherosclerotic lesions from the aorta up to the maxillary artery; (2) localization of atherosclerotic lesions with a lower CT content in the diet is dependent on the duration of feeding and may involve the aorta up to the external carotid artery; (3) the development of the atherosclerotic lesion in hypercholesterolemic rabbit is strictly related to the appearance of an intermediate SMC subtype; (4) atherosclerotic lesions occur only in those arterial sites which, in corresponding normocholesterolemic rabbit, contain intermediate-type SMC; and (5) no differences can be found in the distribution of SMC subpopulations present in the lesions from WHHL, CT-fed animals, or at various arterial levels, whereas some discrepancies can be shown in aortic atherogenesis. PMID- 7488336 TI - Removal of LDL from plasma by adsorption reduces adhesion molecules on mononuclear cells in patients with arteriosclerosis obliterans. AB - BACKGROUND: There is increasing evidence that immune processes are important in the development of atherosclerosis. We investigated whether low density lipoprotein (LDL) adsorption therapy affected serum cytokine levels and the expression of adhesion molecules on peripheral blood mononuclear cells (lymphocytes and monocytes) in patients with arteriosclerotic obliterance (ASO). METHODS AND RESULTS: LDL adsorption therapy was repeated ten times over a period of three months in ten ASO patients. The total serum cholesterol and LDL cholesterol levels were significantly reduced at the end of therapy. This was associated with a significant improvement in Fontaine's classification and ankle pressure index. We also measured serum levels of inflammatory cytokines (interleukin-1 beta (IL-1 beta), IL-6 and tissue necrosis factor alpha (TNF alpha)) and expression of adhesion molecules (lymphocyte function-associated antigen 1 alpha (LFA-1 alpha), LFA-1 beta, CD2, very late antigen (VLA)-4, VLA-5 and CD44) on mononuclear cells in the same patients and a group of healthy subjects. Serum levels of all inflammatory cytokines were markedly higher in ASO patients compared with healthy subjects, but there was no significant difference in the level before and after LDL adsorption. VLA-4 expression on CD3+ cells, but not of other adhesion molecules, was markedly higher in ASO patients compared with healthy subjects. LDL adsorption caused a significant reduction in CD2, VLA4 and VLA-5 expression on CD3+ cells. Furthermore, VLA-4 and VLA-5 expression on monocytes diminished significantly after LDL adsorption. CONCLUSIONS: Our results indicate that LDL adsorption-induced immunoregulation is mediated by an indirect stimulatory effect on the immune system. The results suggests that improved peripheral circulation produced by LDL adsorption may reflect improved immune dysfunctions of atherosclerotic lesions in ASO patients. PMID- 7488337 TI - Soluble interleukin-2 receptor as a clinical parameter for nasopharyngeal carcinoma. AB - We evaluated whether the serum soluble interleukin-2 receptor (sIL-2R) may be a parameter to monitor the efficacy of treatment for nasopharyngeal carcinoma (NPC). There were 177 NPC patients and 24 healthy controls. The level of sIL-2R was measured with a sandwich ELISA kit. Higher levels of sIL-2R than for controls were found in NPC patients before treatment and in patients with distant metastasis (p < 0.001). There was, however, no difference in sIL-2R levels between controls and NPC patients after radiotherapy in relapse-free or in primary relapse. The sIL-2R levels in sequential testing revealed good correlation with clinical response. The sIL-2R levels were found to be elevated when distant metastasis was detected. Two patients had elevated sIL-2R level up to 5 months before clinical detection of metastasis. These results indicate that serial measurements of sIL-2R levels are worthwhile for NPC patients in their clinical course. The sIL-2R level proved to be an adjunct clinical parameter to monitor the efficacy of treatment of NPC. PMID- 7488338 TI - Prevalence of allergy to some inhalants among rhinitis patients in Malaysia. AB - This study was conducted to determine the seasonal prevalence of allergies to house dust, D. pteronyssinus, D. farinae, cat fur, dog hair, mixed moulds, mixed grass pollens and American cockroach. A total of 314 patients with clinically suspected allergic rhinitis was examined by prick test using commercial preparations of the above allergens. Total serum IgE of the patients was determined by a Sandwich ELISA. Ninety-six percent of the patients tested positive to more than one allergen. Most were positive to a combination of 4 allergens. More than 70% of the patients were positive to house dust, D. pteronyssinus, D. farinae and cat fur. Analysis indicates that for an individual who tests positive for house dust, there is a very high risk of the person being allergic to the dust mites and cat fur too. Most of the allergens had 2 peak period of high positive PT rates; mixed moulds and mixed grass pollens had 3 peaks. There was significant positive correlation between the monthly positive PT rates against mixed moulds and mixed grass pollens with maximum daily mean temperature and mean temperature at 14.00 hours. PMID- 7488341 TI - Cold urticaria in Thai children: comparison between cyproheptadine and ketotifen in the treatment. AB - The study was performed in 6 Thai children with primary acquired cold urticaria. They all suffered from generalized urticaria and two of them also had angioedema. All of them had normal erythrocyte sedimentation rate, complement 3 and negative VDRL, TPHA, hepatitis B screen and cold agglutinin titer. Cryoglobulin was checked in 3 cases and showed negative results A double-blind cross-over study to compare the effectiveness of cyproheptadine and ketotifen demonstrated that the efficacy of cyproheptadine and ketotifen on clinical symptoms and ice cube test was not significantly different (p > 0.05). Both of them showed good results in the treatment of cold urticaria with mild side effects. During the follow up, 5 cases showed complete recovery while the other one developed one or two exacerbations per year upon cold exposure. However, the symptoms were mild and subsided on administration of one or two doses of H1 antihistamine. Our data demonstrated that ketotifen was as effective as cyproheptadine in the treatment of cold urticaria in Thai children. PMID- 7488339 TI - Seasonal prevalence of air-borne pollen and spores in Kuala Lumpur, Malaysia. AB - Aerosampling using Rotorod samplers was conducted in the Institute for Medical Research, Kuala Lumpur, Malaysia, from December 1991 to November 1993. Samples were collected twice a week between 10.00 hours to 12.00 hours. Rods were stained and examined microscopically. A total of 8 and 20 types of pollens and mold spores were collected, respectively. More mold spores were collected than pollens. Grass pollen constituted more than 40 percent of total pollen counts. Gramineae pollen counts peaked in March and September. The most abundant mold spore was Cladosporium followed by Rust, Nigrospora, Curvularia and Smut. Cladosporium counts peaked in February and August. Rust counts peaked in June and December whereas counts for Nigrospora peaked in February and October. Highest counts of Smut were recorded in March and October. Curvularia counts peaked in January, June and September. PMID- 7488340 TI - Specific IgE in the identification of allergens in allergic rhinitis Malaysian patients. AB - The specific serum IgE levels to 20 allergens were determined by enzyme immunoassay in 90 Malaysian patients with allergic rhinitis. Ninety-two percent of patients had elevated IgE to at least 1 of the allergens. The housedust mites D. pteronyssinus and D. farinae were the major allergens, elevated IgE to either allergen being present in 86% of the patients. Prick skin tests were carried out in some of the patients, housedust mites, cat fur, dog hair and shrimp were the allergens used. Close correspondence was found between IgE and prick skin tests to the mites. PMID- 7488342 TI - Serological response to respiratory syncytial virus infection in pediatric patients with a comparison to immunofluorescence and virus isolation. AB - The serological response to respiratory syncytial virus (RSV) in 125 pediatric patients hospitalized with acute lower respiratory infection was investigated by enzyme linked immunosorbent assay (ELISA) for specific immunoglobulin (Ig) A, IgG, and IgM and complement fixation (CF) test. By ELISA, a 4-fold rise in IgG titre in paired sera was most commonly found, followed by a rise in IgA and IgM titres. Investigation by ELISA and CF leads to the suggestion that major CF activity against RSV antigens resides in the IgG and not the IgA and IgM classes. No case with CF activity failed to be diagnosed by ELISA. The youngest infant who could develop seroconversion was one month old, nevertheless two children older than two years could not. When the three diagnostic methods were compared, ELISA serology was the most sensitive followed by indirect immunofluorescence (IIF) for antigen detection and virus isolation, respectively, ELISA could diagnose RSV infection in 45% of the study cases, whereas IIF and virus isolation only diagnosed 26% and 14%, respectively. Half of the cases was diagnosed by all of the three methods together. PMID- 7488343 TI - Radioactive and non-radioactive lymphocyte proliferation assays for measuring rabies-specific cellular immunity. AB - We compared the radioactive lymphocyte proliferation assay for measuring rabies specific cell-mediated immunity in a group of 42 rabies vaccine recipients with a new technique using rabies-stimulated lymphocytes in a colorimetric test kit (CellTiter 96, Promega Corporation, USA). Results of both tests were good agreement (Kappa = 0.68), however, they did not show good correlation in degree (magnitude) of positivity. In addition, the conventional assay showed a higher degree of sensitivity. PMID- 7488344 TI - Establishment of OVS1 and OVS2 monoclonal antibodies recognizing human ovarian mucinous cystadenocarcinoma. AB - Two newly established murine monoclonal antibodies (MAbs), OVS1 and OVS2, to human ovarian mucinous cystadenocarcinoma were further characterized for diagnostic efficacy. The specific SA-1 antigen, purified from the tumor extract was identified as a glycoprotein of 29 kDa. A double determinant biotinstreptavidin alkaline phosphatase immunoassay system, containing OVS1 and OVS2 MAbs was used to determine the SA-1 levels in serum. The OVS1 MAb was used as a first antibody because of its high specificity of 96% while OVS2 MAb, with a lower specificity of 8% but greater sensitivity of 78%, was chosen as a second antibody. Matched sera of 64 healthy controls and 90 patients with definite diagnoses of 25 benign diseases, 14 nonovarian cancer and 51 ovarian cancer, were simultaneously measured together with CA 125 values. At cut-off levels of 220 and 360 units/ml, the SA-1 test showed 63% and 43% positive rates respectively in all types of ovarian cancer, compared to 65% and 57% positive rates for CA 125 at cut off levels of 35 and 60 units/ml, respectively. Sensitivity for SA-1 at 220 units/ml cut-off level in mucinous ovarian cancer was 75% and increased significantly to 85% when the test was combined with CA 125 at 35 units/ml cut off level. Furthermore, The combination of both tests significantly increased the positive rates to 86% in all types of early stage ovarian cancer.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7488345 TI - Methacholine inhalation challenge in patients with post-Mycoplasma pneumoniae pneumonia. AB - We studied methacholine bronchial inhalation challenge in 12 patients at 4th week and 12th week after recovered from Mycoplasma pneumoniae pneumonia, compared with 12 healthy subjects as controls. The aerosolized methacholine was produced by an atomized nebulizer of the Provocationtest I, Pari-Starnberg, Germany and the aerosol was kept into a reservoir bag. Then, it was inhaled slowly by a subject. Increasing concentration of methacholine solutions (0, 0.5, 1, 5, 10, and 25 mg/ml) were used. The results revealed that 67% of the patients had bronchial reactivity to methacholine at the first time of challenge with a mean concentration of methacholine producing a fall in FEV1 of 20% from baseline (PC20) of 12.3 +/- 6.44 mg/ml. Fifty percent of the patients were still positive to the test on the second time of challenge with a mean PC20 of 20.1 +/- 6.89 mg/ml. None of the healthy subjects had bronchial hyperreactivity (PC20 > 25 mg/ml). Two patients experienced wheezing and asthmatic attacks requiring bronchodilator therapy during acute phase pneumonia. They were also diagnosed as having bronchial asthma for the first time. Many patients had prolonged coughing during the recovery phase lasting more than 4 weeks. This prolonged coughing seemed to have a correlation with the development of bronchial hyperresponsiveness (BHR). We concluded that M. pneumoniae could induce BHR which may be transient or persistent. The effect of mycoplasma respiratory tract infection may result in airway inflammations and asthmatic attacks. PMID- 7488346 TI - Production of monoclonal antibody specific to Campylobacter jejuni and its potential in diagnosis of Campylobacter enteritis. AB - A monoclonal antibody (MAb 3G6) specific for Campylobacter jejuni and Campylobacter coli was produced by immunizing BALB/c mice with a local strain of C.jejuni (28.1). No cross-reactivity was observed with Enterobacteriaceae controls. By immunoprecipitation, MAb 3G6 identified a major protein band of molecular weight 45 kDa and also gave a slight reactivity with 30 and 55 kDa proteins. Using an indirect enzyme-linked immunosorbent assay, MAb 3G6 was able to detect C.jejuni suspended in stool without cross-reactivity to 14 other enteropathogenic bacteria suspended, normal flora in fecal suspension, or fecal debris. In the analysis of fifty clinical specimens, MAb 3G6 detected most positive samples with the exception of one which possessed very low Campylobacter concentration and gave no reactivity to negative samples, demonstrating its high specificity to C. jejuni and C. coli. MAb 3G6 may be suitable as a new tool for the development of diagnostic method for Campylobacter infection. PMID- 7488347 TI - Monoclonal antibodies against protein antigens of salmonellae causing paratyphoid fever and their diagnostic application. AB - Hybrid clones producing monoclonal antibodies (MAbs) specific for Salmonella paratyphi A (72 clones), S. paratyphi B (9 clones) and S. paratyphi C (8 clones) were produced by using the affinity purified Salmonella protein (Bp) as immunogens. MAbs to S. paratyphi A and S. paratyphi B reacted specifically with the 52 kDa homologous flagellin protein components while those to S. paratyphi C reacted with a 61 kDa flagellin protein component. The MAbs against S. paratyphi A and S. paratyphi B were used to establish a double antibody sandwich ELISA for detection of the 52 kDa flagellin antigens in serum samples from patients with acute paratyphoid A and paratyphoid B fever. With this assay system, 6.25 ng per ml of flagellin antigens of S. paratyphi A and S. paratyphi B could be detected. However, the assay system could not detect the flagellin antigens in patients' sera. The presence of IgM antibodies to the 52 kDa antigens of S. paratyphi A and S. paratyphi B in the acute sera from paratyphoid A or paratyphoid B patients suggested that the 52 kDa protein components of both salmonellae are good immunogens for human and might be used as antigens for early diagnosis of paratyphoid A and paratyphoid B fever. PMID- 7488348 TI - Cloning of Thai strain Giardia intestinalis. AB - Axenic cultures of Giardia intestinalis trophozoites were successfully established from human fecal specimens and rectal swabs from dogs using sucrose gradient centrifugation to separate the cysts from fecal material, the excystation method of Robert-Thompson et al. and culture of the preparation in TYI-S-33 medium supplemented with human serum, vitamin mixture and piper acillin and amikacin antibiotics, respectively. Fungal contamination could be controlled by amphotericin B at 10 micrograms per ml of medium. Clones of the parasites were obtained using a combination of dilution method and micromanipulation technique. PMID- 7488349 TI - Characterization of immunological memory cells. A minireview. AB - This paper reviews the information concerning immunological memory T and B cells. It shows that the existence of different or a single lineage of memory and naive cells is still a question. The recirculation pattern of memory cells is different from naive cells. A unique recirculation for memory T or B cells is suggested while the bcl2 gene expression possibly plays role in the longevity of memory cells. Various phenotypic markers are demonstrated only on memory or on naive cells; however, a reliable and convenient method for the detection of memory cells still needs to be explored. PMID- 7488350 TI - Use of globic scattering factors for protein structures at low resolution. AB - At 3 to 4 A resolution, the electron density of a protein may be modeled by a continuous chain of 'globs' representing the amide region of the peptide backbone and the side-chain residues. Group scattering factors are derived from a trans planar C alpha C = ONC alpha backbone segment and most favored side-chain conformer for 18 different amino acids. Trial calculations indicate that the phase error and crystallographic residual comparing the atomic and 'globic' models rapidly decrease from high to low resolution. At 3 A resolution, the phase error is approximately 80 degrees. These results indicate that the electron density of a protein composed of N amino acid residues may be adequately modeled by 2N globs at low resolution. PMID- 7488351 TI - International Scientific Assembly on the Comparative Anticaries Efficacy of Sodium Fluoride and Sodium Monofluorophosphate Dentifrices. Proceedings. London, England, August 18, 1993. PMID- 7488352 TI - A critical review of the 10 pivotal caries clinical studies used in a recent meta analysis comparing the anticaries efficacy of sodium fluoride and sodium monofluorophosphate dentifrices. AB - The 10 pivotal caries clinical studies employed in a recent meta-analysis to compare the anticaries efficacy of sodium fluoride (NaF) and sodium monofluorophosphate (MFP) dentifrices were subjected to a critical review. When reporting the meta-analysis, the author (Johnson) considered these 10 pivotal caries clinical studies as "head-to-head" comparisons of NaF and MFP dentifrices. For the most part, it is difficult to find any common denominator among these 10 caries clinical studies since there were so many diverse factors associated with them. The 10 pivotal studies were conducted over a 20-year period of time in many different geographic areas of the world, using different protocols and study designs, different clinical examiners and caries assessment criteria, as well as evaluating commercially outdated or specially formulated dentifrices. Any attempt to integrate the results from these 10 studies is further complicated by the diversity of statistical methodology used to evaluate the results obtained from the studies. Also, in two of the 10 studies, the comparison of the fluoride dentifrices was obscured by the fact that the participating children used a fluoride mouthrinse during the course of the study. In any case, when all relevant dentifrice comparisons are made, the results of a critical review of the 10 pivotal ("head-to-head") caries clinical studies indicate the following: (1) In three of the studies (Gerdin/Zacherl/Forsman), depending upon how the data is viewed, it can either be shown that NaF dentifrices are favored over MFP dentifrices or that MFP dentifrices are favored over NaF dentifrices. (2) In one of the studies (Edward & Torell), the results are published only in abstract form and a full report of the study is not available. (3) In one of the studies (Koch, Petersson, Kling & Kling) it is reported that MFP dentifrices are favored over NaF dentifrices. (4) In two of the studies (Edlund & Koch, Koch et al) it is reported that NaF dentifrices are favored over MFP dentifrices. However, in both of these studies, there were serious imbalances in baseline caries status favoring the NaF dentifrice groups over the MFP dentifrice groups. In other words, the children using the MFP dentifrices were more prone to caries formation during the course of these studies than the children using the NaF dentifrices. Further, in these two studies there is a serious question regarding the soluble fluoride level of the commercially available MFP dentifrice that was used.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 7488353 TI - Clinical significance of the conclusions from the International Scientific Assembly on the Comparative Anticaries Efficacy of Sodium Fluoride and Sodium Monofluorophosphate Dentifrices. PMID- 7488354 TI - Statistical considerations related to a meta-analytic evaluation of published caries clinical studies comparing the anticaries efficacy of dentifrices containing sodium fluoride and sodium monofluorophosphate. AB - The comparison of the anticaries efficacy of dentifrices containing sodium fluoride (NaF) and sodium monofluorophosphate (SMFP) has recently been addressed through a meta-analysis of published head-to-head clinical trials. Such an analysis is used to provide an omnibus conclusion on the basis of summary information obtained from several individual studies. An important aspect of a meta-analysis is the determination of which studies from among those available in the relevant literature are to be incorporated into the calculations. The statistical literature provides several perspectives with respect to this issue. A recently-published meta-analysis by Johnson (1993) resulted in the conclusion that dentifrices containing NaF provide significantly lower caries increments than do those containing SMFP. However, a similar analysis utilizing studies identified by Volpe, Petrone & Davies (1993) suggests that no such significant difference exists. The resolution of such conflicting results rests on clinical judgment concerning the issue of study inclusion. The clinical significance of the results of both of these analyses was considered. Despite the differences in the analyses, both support the conclusion that dentifrices containing SMFP provide equivalent anticaries protection as do those containing NaF, in accordance with the standards established in the current American Dental Association Guidelines for the comparison of fluoride dentifrices. PMID- 7488355 TI - The International Scientific Assembly on the Comparative Anticaries Efficacy of Sodium Fluoride and Sodium Monofluorophosphate Dentifrices. PMID- 7488356 TI - Critique of Kingman's methods of projecting long-term relative efficacy of products exhibiting small short-term efficacy. AB - This paper critically reviews Kingman's "methods of projecting long-term relative efficacy of products exhibiting small short-term efficacy," a report he prepared for a scientific workshop on the relative anticaries efficacy of fluoride dentifrices, which reveals serious deficiencies. Of the three specific models that Kingman suggests, the compound growth model (CGM), which leads to the largest projected future percentage reductions, is based on a false premise of an increasing universe of caries-free surfaces available for caries-attack with the passage of time. The second suggested model, the demineralization square root model (DSR), is supported only by a personal communication, unavailable for scrutiny. The third suggested model, a stabilization model (STA) is plausible, but unsupported. Other plausible models for extrapolating short-term observed effects are ignored by Kingman. Most of Kingman's choices of data illustrations to fit his suggested models are ill-chosen because the studies were repeated cross-sectional surveys or involved the delivery of systemically and topically delivered fluorides, which contraindicate their use. The most serious breach is Kingman's selective use of subsets of data from two long-term studies of fluoride dentifrices that support his premise, when other, more comprehensive data from the same studies do not. He also ignores findings from other studies that fail to support his suggested models. Kingman's report fails to prove that studies which show small short-term effectiveness after 1, 2 or 3 years will lead to greater relative effectiveness after long periods of use. PMID- 7488357 TI - Sodium fluoride or sodium monofluorophosphate? A critical view of a meta-analysis on their relative effectiveness in dentifrices. AB - A detailed examination of nine publications used in a meta-analysis to compare the relative effectiveness of sodium fluoride and sodium monofluorophosphate in dentifrices showed important flaws in the process. Some studies were inappropriately chosen. Other selected comparisons between the two compounds favored sodium fluoride when more appropriate comparisons in the same studies, showing either an advantage for sodium monofluorophosphate or a lesser advantage to sodium fluoride, were ignored. Some simple errors also appear to have been made when taking data from the publications. When all these factors are taken into account it seems that three studies favor sodium monofluorophosphate, two favor sodium fluoride and the remaining five either could not or should not be included in a meta-analysis. The only two scientifically conceived and conducted studies failed to demonstrate an advantage of one compound over the other. PMID- 7488358 TI - A comprehensive summary of fluoride dentifrice caries clinical trials. PMID- 7488360 TI - Enzyme reactions in the presence of cyclodextrins: biosensors and enzyme assays. AB - Cyclodextrins, macrocyclic carbohydrates with apolar internal cavities, can form complexes with, and solubilize many normally water-insoluble compounds. Ferrocene and its derivatives, tetrathiafulvalene and tetramethylbenzidine, can function as redox mediators, but are insoluble in water; when they are complexed with cyclodextrins, they can be used in enzymatic assays and in the construction of mediated biosensors. In addition, the solubilization of polynuclear aromatic hydrocarbons (PAHs), including the potent carcinogen benzo[a]pyrene, by cyclodextrins has enabled the detection of these important environmental contaminants. PMID- 7488359 TI - The relative anticaries effectiveness of sodium monofluorophosphate and sodium fluoride as contained in currently available dentifrice formulations. AB - A 36-month double-blind clinical caries study was conducted to determine the comparative anticaries efficacy of sodium monofluorophosphate (MFP) and sodium fluoride (NaF) as incorporated in dentifrice formulations. The test dentifrices which were utilized are commercially available in the United States and all have been granted the American Dental Association Seal of Acceptance for anticaries efficacy. The study was conducted in accordance with the 1988 Guidelines of the American Dental Association Council on Dental Therapeutics. The two formulations used in the NaF versus MFP comparison were Colgate Winterfresh Gel with 0.76% sodium monofluorophosphate in a silica base and Crest Regular Flavor Toothpaste with 0.24% sodium fluoride in a silica base. A third commercially-available NaF dentifrice, Colgate Junior Toothpaste with 0.243% sodium fluoride in a silica base, was included in the study for further comparison. Schoolchildren residing in Maine, U.S.A. received a baseline caries clinical examination and then were randomly assigned to the twice-daily use of one of the three dentifrices which were purchased in the marketplace and repackaged in plain white tubes. Caries clinical examinations were subsequently conducted after 2 and 3 years' use of the dentifrices. A total of 2,222 children participated in both the 2 and 3-year caries clinical examinations. The 36-month DFS (decayed and filled surfaces) and the DFT (decayed and filled teeth) mean caries increments for the three dentifrice groups were virtually identical. The children using the MFP formulation experienced a DFS of 1.50 and a DFT of 1.08; those using the Crest NaF formulation experienced a DFS of 1.48 and a DFT of 1.07; subjects in the third group (Colgate Junior Toothpaste, 0.243% NaF) had a DFS of 1.49 and DFT of 1.06. The statistical evaluation of the data consisted of the calculation of 90% confidence bounds for the ratio of (true) mean increments, in accordance with the 1988 American Dental Association Guidelines. Expressed in the language of the Guidelines, the analyses indicated that, (1) the anticaries efficacy provided by the MFP formulation, (Colgate Winterfresh Gel with 0.76% MFP) is "as good as" that provided by the NaF formulation, (Crest Regular Flavor Toothpaste, with 0.243% NaF), and (2) the anticaries efficacy provided by the one NaF formulation (Colgate Junior Toothpaste, with 0.243% NaF) is "as good as" that provided by the other, (Crest Regular Flavor Toothpaste, with 0.243% NaF). PMID- 7488361 TI - Binaural function in children with attention-deficit hyperactivity disorder. AB - OBJECTIVE: To evaluate the basic binaural function of children with attention deficit hyperactivity disorder (ADHD). DESIGN: Case-control study. Measures of binaural function were compared for children with and without ADHD. SUBJECTS: Forty-two children; 26 had ADHD and 16 were normal controls. INTERVENTIONS: None. RESULTS: For detection tasks, no differences were seen between children with ADHD and controls. For speech recognition tasks, the younger children with ADHD did not perform as well as the controls. CONCLUSIONS: Younger children with ADHD might have a reduced processing efficiency for signal recognition in certain types of noise, but not for signal detection. PMID- 7488362 TI - Unilateral endoscopic supraglottoplasty for severe laryngomalacia. AB - OBJECTIVE: To evaluate the effectiveness of unilateral supraglottoplasty in the treatment of children with severe laryngomalacia. DESIGN: Retrospective study. SETTING: Pediatric tertiary referral center. PATIENTS: Eighteen children with severe laryngomalacia. Indications for surgical intervention were obstructive apnea, failure to thrive, cyanosis, and/or cor pulmonale. INTERVENTIONS: Unilateral carbon dioxide laser removal of redundant supraglottic tissue (supraglottoplasty). OUTCOME MEASURES: Evaluation of relief of symptoms, need for subsequent contralateral procedure, and incidence of complications. RESULTS: Three patients required treatment of the opposite side at a later date. There were no complications. Obstructive apnea and weight gain improved in all. CONCLUSIONS: Unilateral supraglottoplasty can be used to treat severe laryngomalacia in most patients. A small percentage of patients will subsequently require a contralateral procedure. Unilateral supraglottoplasty may have less risk of complications than bilateral supraglottoplasty. PMID- 7488363 TI - Extraluminal laryngotracheal fixation with absorbable miniplates. AB - OBJECTIVE: To determine if miniplates made of polydioxanone can be used successfully for extraluminal laryngotracheal fixation. DESIGN: Twelve rabbits underwent anterior and posterior laryngotracheofissure with extraluminal fixation of the divided segments with polydioxanone miniplates. Three control animals did not undergo plating. Animals were killed after 2, 4, and 12 weeks. Larynges were removed and examined to determine the intraluminal area and histologic status of the repair site. RESULTS: The intraluminal area of the animals in the experimental group was 1.6 times that of those in the control group. The trachea was relined with respiratory mucosa by 2 weeks. Little inflammatory reaction was evident. Plate resorption was almost complete at 12 weeks. CONCLUSION: Polydioxanone plating is a feasible and well-tolerated method of extraluminal laryngotracheal fixation. It may be useful in the treatment of grade 2 or 3 subglottic stenosis. PMID- 7488364 TI - The effects of selective nerve stimulation on upper airway airflow mechanics. AB - OBJECTIVE: To evaluate the effect of electrical stimulation of hypoglossal nerve branches and ansa cervicalis nerve branches on upper airway patency. DESIGN: Pressure-flow relationships obtained during supramaximal stimulation of hypoglossal nerve branches and ansa cervicalis nerve branches were analyzed in the isolated feline upper airway to determine the maximum inspiratory airflow as well as to determine pharyngeal collapsibility (upper airway critical pressure) and nasal resistance upstream from the site of pharyngeal collapse. Comparisons were performed between baseline and stimulation conditions with paired two-tailed t tests. RESULTS: Stimulation of the proximal hypoglossal nerve trunk, distal medial hypoglossal nerve branch, nerve branches to the suprahyoid muscles, the infrahyoid muscles, and the suprahyoid and infrahyoid muscles simultaneously increased maximum inspiratory airflow significantly by decreasing airway collapsibility. A greater reduction in airway collapsibility was observed with stimulation of the tongue muscles compared with stimulation of the strap muscles. CONCLUSIONS: Stimulation of specific hypoglossal and ansa cervicalis nerve branches consistently increased maximum inspiratory airflow by decreasing airway collapsibility. The major decrease in airway collapsibility from hypoglossal nerve stimulation is dependent on the action of the genioglossus muscle. PMID- 7488365 TI - Midfacial fractures in pediatric patients. Frequency, characteristics, and causes. AB - OBJECTIVE: To determine the frequency, characteristics, and causes of midfacial fractures in children. DESIGN: A retrospective review of the patients' medical charts and radiographs. SETTING: Tertiary referral center. PATIENTS: Fifty-four patients under 16 years of age, with midfacial fractures diagnosed and treated in the Helsinki (Finland) University Central Hospital from 1980 through 1992. MAIN OUTCOME MEASUREMENTS: The data examined included sex, age, time and cause of the accident, type and location of the fractures, the presence and location of associated injuries, complications, and treatment methods. RESULTS: The male-to female ratio was 1.16:1. Motor-vehicle accident was the most common cause of injuries. The frequency of injuries was in decreasing order: (1) maxillary alveolar bone, (2) zygoma, and (3) Le Fort fractures of the maxilla. The majority of injuries occurred in subjects who were 13 to 15 years old. In children less than 6 years old, only alveolar fractures occurred. For the other age groups, no significant difference in the fracture pattern was found. No fatalities occurred in this patient series. CONCLUSIONS: Midfacial pediatric fractures are rare. A high velocity force, such as that from a motor-vehicle accident is a factor producing the injury. Owing to the high impact, associated injuries are common. The severity of the insult is more essential than the age of the patient and the development of the paranasal sinuses. PMID- 7488366 TI - Orbital fractures in children. AB - OBJECTIVE: To determine if the pattern of orbital fractures may be influenced by the changing craniofacial ratio of the growing child, as the orbit is the boundary between the face and the cranium. DESIGN: Retrospective case series of 40 patients between the ages of 1 year and 16 years with orbital fractures. SETTING: The Albany (NY) Medical Center Hospital, a tertiary level 1 trauma center. OUTCOME MEASURES: The sex, age, site, and mechanism of injury, associated injury, and treatment methods for children admitted to the Albany Medical Center Hospital with orbital fractures between July 1986 and June 1992. RESULTS: Fourteen children had fractures of the orbital roof, 10 children had fractures of the orbital floor, 14 children had mixed fractures, and two children had fractures of the medial wall. The mean age (4.8 +/- 3.3 years) of the 14 patients with roof fractures was significantly less than the mean age (12.0 +/- 4.2 years) of the 26 children with other orbital fractures. Logistic regression demonstrated that the age at which the probability of lower orbital fractures exceeds the probability of orbital roof fractures is 7.1 +/- 1.0 years. Orbital roof fractures had a significantly greater likelihood of associated neurocranial injuries. The need for surgical repair was significantly lower among children with roof fractures as well as among children 7 years of age and younger. CONCLUSIONS: Orbital roof fractures are a type of skull fracture that occur primarily in younger children as a consequence of the proportionally larger cranium and the lack of frontal sinus pneumatization. Lower orbital fractures are a type of facial fracture that occur primarily in older children as a consequence of the increased vulnerability of the face due to growth and the pneumatization of the paranasal sinuses. PMID- 7488367 TI - Management of posterior laryngeal and laryngotracheoesophageal clefts. AB - OBJECTIVE: To review the clinical features, associated congenital abnormalities, management, and morbidity of infants presenting with posterior laryngeal and laryngotracheal clefts. DESIGN: Case series. SETTING: Great Ormond Street Hospital for Sick Children NHS Trust, London, England. PATIENTS: Consecutive sample of 44 patients presenting with posterior laryngeal and laryngotracheal clefts between December 10, 1979, and January 30, 1992. MAIN OUTCOME MEASURES: Clinical features, incidence of surgery, and associated morbidity and mortality related to different types of airway cleft. RESULTS: The main presenting features were stridor and aspiration, which were more evident with the more extensive clefts. Twenty-five patients (56%) had associated congenital abnormalities. Fourteen patients (32%) were treated conservatively. Sixteen patients (36%) underwent primary endoscopic surgical repair. Eight patients (18%) underwent primary repair via an anterior laryngofissure; and six patients (14%) underwent primary repair via a lateral pharyngotomy. Eight patients (18%) required revision surgery, two (4%) of them on more than one occasion. Ten patients (23%) required fundoplication to control gastroesophageal reflux. Six patients (14%) died. CONCLUSIONS: The identification of an airway cleft requires a high index of suspicion. Morbidity and mortality are reduced by securing the airway, controlling gastroesophageal reflux, and using a multidisciplinary pediatric team. We recommend the anterior laryngofissure because of the ease of surgical access. PMID- 7488368 TI - Task force on recurrent respiratory papillomas. A preliminary report. AB - OBJECTIVE: To obtain pilot data about the incidence, need for surgical intervention, and demographics of recurrent respiratory papillomas in the United States. DESIGN: Otolaryngologists were surveyed using a questionnaire with structured and open-ended questions. PARTICIPANTS: One thousand board-certified otolaryngologists practicing in the United States as of January 1, 1993, through a random mailing list provided by the American Academy of Otolaryngology-Head and Neck Surgery and all active US members of the American Society of Pediatric Otolaryngology and the American Bronchoesophagological Association. A total of 1346 questionnaires were distributed. MAIN OUTCOME MEASURES: Physician's responses to questions about their current patient load of children and adults with recurrent respiratory papillomas, their surgical and anesthetic management of the disease, and their clinical experiences with risk factors for developing recurrent respiratory papillomas. RESULTS: Projected totals for recurrent respiratory papillomas among children were 2354 new cases (95% confidence interval [CI], 1448 to 3260) and 5970 active cases (95% CI, 3465 to 8474), requiring 16,597 surgical procedures (95% CI, 6938 to 26,255) at a cost of $109 million (95% CI, $45 to 172 million) for March 1, 1993 to March 31, 1994. Projected totals for adult recurrent respiratory papillomas were 3623 new cases (95% CI, 2359 to 4887) and 9015 active cases (95% CI, 6435 to 11,591) requiring 9284 surgical procedures (95% CI, 6003 to 12,565) at a cost of $42 million (95% CI, $27 to $59 million) for March 1, 1993 to March 31, 1994. The incidence of recurrent respiratory papillomas among children is estimated at 4.3 per 100,000; among adults, 1.8 per 100,000. The carbon dioxide laser was favored by 92% of respondents. No consensus was reached on the role for cesarean section. Forty-six percent of respondents favored the use of a laser-safe endotracheal tube for anesthetic management. CONCLUSION: A registry of patients with recurrent respiratory papillomas would benefit future research protocols and provide long term follow-up of patients. PMID- 7488369 TI - Peripheral primitive neuroectodermal tumors of the head and neck. AB - OBJECTIVE: Primitive neuroectodermal tumor is a malignant small round cell tumor of neuroectodermal origin. The occurrence of these tumors in the head and neck is rare. Our objective is to document the frequency of presentation, methods of diagnosis, and forms of treatment used to combat primitive neuroectodermal tumors in the head and neck. DESIGN: Retrospective chart review. RESULTS: The most common location for primitive neuroectodermal tumors in our series was the thoracopulmonary region (46%); the next most common location was the head and neck (42%). Metastatic disease was present in 31% of patients at the time of diagnosis. Cytogenic analysis was found to be helpful in confirming the diagnosis of primitive neuroectodermal tumor. Therapy for primitive neuroectodermal tumors included surgery, chemotherapy, and radiation therapy. Survival statistics were poor, with 65% of the patients alive at 2 years' follow-up. CONCLUSIONS: Primitive neuroectodermal tumor is an aggressive malignant small round cell tumor that may present in the head and neck. Long-term survival for patients with primitive neuroectodermal tumor is still poor; however, the use of improved chemotherapeutic agents combined with aggressive surgical control of primary disease and, in some cases, radiation therapy should lead to improved long-term survival. PMID- 7488370 TI - Eosinophilia in chronic childhood sinusitis. AB - OBJECTIVES: To quantify eosinophilia in sinus tissues obtained from children with chronic sinusitis and to correlate the degree of eosinophilia with history of asthma, allergy, cystic fibrosis, and preoperative computed tomographic (CT) scans. DESIGN: Examination of surgical specimens from children who underwent functional endoscopic sinus surgery and controls. SETTING: Tertiary care medical center. PATIENTS: Thirty-four children who underwent functional endoscopic sinus surgery for chronic sinusitis refractory to medical treatment were divided into three groups: 13 with asthma, 11 without asthma, and 10 with cystic fibrosis. Normal sphenoid sinus mucosa was also obtained from six adults undergoing transsphenoidal hypophysectomies. MAIN OUTCOME MEASURES: Number of lamina propria and intraepithelial eosinophils in surgical specimens, allergic status, presence or absence of asthma, and CT scans obtained preoperatively. RESULTS: There were significantly more lamina propria and intraepithelial eosinophils in the tissue of children with chronic sinusitis compared with normal sphenoid sinus mucosa. More eosinophils were counted in the tissues of patients with asthma and cystic fibrosis compared with patients without concomitant disease, but this did not reach statistical significance. Allergy status did not affect the degree of tissue eosinophilia. Eosinophilia did not correlate with severity of mucosal disease as assessed by CT scans. CONCLUSIONS: Tissue eosinophilia is a characteristic histologic feature of chronic sinusitis in children, especially those with asthma. The presence of allergy does not predict tissue eosinophilia. Furthermore, the degree of tissue eosinophilia does not correlate with the severity of mucosal thickening seen on CT scans. PMID- 7488371 TI - Role of bacterial interference and beta-lactamase-producing bacteria in the failure of penicillin to eradicate group A streptococcal pharyngotonsillitis. AB - OBJECTIVE: To determine the association among bacterial interference and beta lactamase production and penicillin failure in treating streptococcal pharyngotonsillitis. DESIGN: Fifty-two children who had acute pharyngotonsillitis caused by group A beta-hemolytic streptococci (GABHS) were treated for 10 days with penicillin. Surface tonsillar cultures were obtained before therapy and at 10, 21, and 42 days after termination of therapy. The cultures obtained before and 10 days after completion of treatment were processed for aerobic and anaerobic organisms; the other cultures were processed for GABHS only. RESULTS: Based on eradication of GABHS, 38 patients were in the classification bacteriologic "cure"; 14 were in the classification bacteriologic "failure" after therapy. In the cured group, before therapy alpha-hemolytic streptococci inhibiting their own GABHS were recovered in the cultures of 14 children (37%), and beta-lactamase-producing organisms (BLPB) were detected in the cultures of two children (5%). After therapy, inhibiting alpha-hemolytic streptococci were recovered in 31 cultures (82%), and BLPB were detected in five cultures (13%). In contrast, in the failure group, before therapy alpha-hemolytic streptococci were isolated in one culture (7%) and BLPB were recovered from nine cultures (64%). After therapy, alpha-hemolytic streptococci were recovered in four cultures (29%), and BLPB was recovered in 13 cultures (93%). CONCLUSIONS: These data show that the absence of interfering alpha-hemolytic streptococci and the presence of BLPB is associated with penicillin failure in the treatment of GABHS pharyngotonsillitis. PMID- 7488372 TI - Distance between the tonsillar fossa and internal carotid artery in children. AB - OBJECTIVE: Obtain normative data on the distance from the tonsillar fossa to the internal carotid artery in children, and relate this distance to age and weight parameters. DESIGN: A prospective study of 100 children undergoing magnetic resonance imaging studies for pathology unrelated to the upper aerodigestive tract. The torus tubarius corresponds to the lateral extent of the tonsillar fossa and was used as the reference site for measurements. One hundred children (aged 7 days to 18 years; median, 6 years) were consecutively entered into the study. Exclusion criteria included any preexisting abnormalities of the upper aerodigestive tract. The study defined the distance from the tonsillar fossa to the internal carotid artery. Relationships of this distance were defined for both age and weight parameters. RESULTS: With increasing age and weight, the distance between the tonsillar fossa and the internal carotid artery increases in a regular fashion to a value approaching 25 mm. Both relationships are exponential with asymptotes being achieved by 56 kg (correlation coefficient, 0.8616; R2 value, 0.7423) or 12 years of age (correlation coefficient, 0.8452; R2 value, 0.7143). CONCLUSION: Age and weight parameters can be used to predict the distance from the tonsillar fossa to the internal carotid artery in children. PMID- 7488373 TI - Treatment of chronic ear disease. Topical ciprofloxacin vs topical gentamicin. AB - OBJECTIVE: To determine and compare the therapeutic efficiency of ciprofloxacin hydrochloride and gentamicin sulfate in the treatment of chronic ear disease. DESIGN: Prospective randomized study. SETTING: Academic tertiary medical center. PATIENTS: Consecutive referred sample of 44 patients with chronic suppurative otitis media randomized into two groups. INTERVENTIONS: Ciprofloxacin hydrochloride (200 mg/mL) was administered to the first group (composed of 24 patients), while the second group (composed of 20 patients) received gentamicin sulfate (5 mg/mL) locally, five drops three times a day for 10 days. RESULTS: In the ciprofloxacin group, 21 (88%) of the 24 patients with suppurative chronic otitis media were cured. On the other hand, only six (30%) of the patients in the gentamicin group were cured. The rest of the patients showed no clinical or bacteriological improvement. CONCLUSIONS: To our knowledge, this is the first study to compare the efficiency of two topical otic preparations in the treatment of chronic ear disease. The results show that topical ciprofloxacin preparation is more efficacious and efficient than topical gentamicin for the treatment of chronic otitis media in the acute stage. PMID- 7488374 TI - Two new otolaryngologic findings in child abuse. AB - We reviewed cases of early childhood hoarseness caused by vocal nodules and functional hearing loss as being possible manifestations of child abuse. This is a retrospective review set in two urban referral centers. The patients consisted of four children with vocal nodules and known histories of abuse, and four children with functional hearing loss and histories suggestive of abuse. Vocal nodules or functional hearing loss may be indicators of an abusive situation. Child abuse should be considered in the differential diagnosis of these problems. PMID- 7488375 TI - Respiratory distress in the neonate. Sequela of a congenital dacryocystocele. AB - Congenital dacryocystoceles presenting with intranasal extension may precipitate respiratory distress in neonates. Twenty-one children have been described as having congenital dacryocystoceles with intranasal extension, and, of these, 14 had respiratory distress. We present a series of patients with congenital dacryocystoceles who presented with ranging degrees of respiratory distress. Magnetic resonance imaging and computed tomographic scans confirmed the diagnosis. In all cases, the endonasal portion of the cyst was marsupialized endoscopically with complete resolution of symptoms. Pertinent embryology, anatomy, clinical presentation, and treatment are discussed. PMID- 7488376 TI - Self-induced pneumoparotitis. AB - Pneumoparotitis is a rare cause of enlargement of the parotid gland; it is often misdiagnosed and therefore incorrectly treated. We report three pediatric cases of self-induced pneumoparotitis and detail the clinical presentation, pathogenesis, radiographic findings, and treatment options. We also review the literature on the subject. In children, inflammatory swelling of the parotid gland is usually due to acute viral or bacterial infection, juvenile recurrent parotitis, or allergic, autoimmune, or systemic disease. Infrequently, swelling may result from air being forced through Stensen's duct, resulting in pneumoparotitis. This may occur as a transient or recurrent phenomenon. Recurrent parotid insufflation is not entirely benign and may predispose to sialectasias, recurrent parotitis, and even subcutaneous emphysema. PMID- 7488377 TI - Pathologic quiz case 2. Oncocytic cyst of the larynx. PMID- 7488378 TI - Pathologic quiz case 1. Microcystic adnexal carcinoma (MAC). PMID- 7488379 TI - Otitis media. PMID- 7488380 TI - Hereditary hearing impairment. PMID- 7488381 TI - Idiopathic perilymphatic fistulas. PMID- 7488382 TI - Fastidious gram-negative bacteria: meeting the diagnostic challenge with nucleic acid analysis. AB - The extended panorama of fastidious Gram-negative bacteria (FGNB) as opportunistic etiological agents of infectious diseases in immunocompromised patients is largely due to improved medical expertise and technology. The heightened awareness of infectious diseases due to FGNB species mandates comprehensive classification and identification systems as a basis for rapid and reliable diagnostics. The most useful approaches are combinations of nucleic acid techniques such as hybridization, genetic transformation, amplification and base sequence analysis with selected conventional criteria. Among these approaches, the widely distributed feature of natural competence in these organisms facilitates the use of the biological method of genetic transformation as a valuable addition to the more common nucleic acid techniques. We describe the development of the taxonomy of FGNB through the last four decades, with particular emphasis on the families Neisseriaceae, Moraxellaceae, and Pasteurellaceae. PMID- 7488383 TI - An electron microscopic study of clinical Paget's disease of the nipple. AB - An ultrastructural study of the epidermis from eight patients with clinical Paget's disease of the nipple supports the epidermotropic theory. There was no evidence that the Paget's cells originated from keratinocytes. We propose the hypothesis that Paget's cells represent transformed ductal cells, from the subjacent lactiferous ducts of the nipple, which have migrated into the epidermis, either as neoplastic cells or as normal ductal cells with secondary in situ transformation in the epidermis. PMID- 7488384 TI - The effect of zinc on bacterial phagocytosis, killing and cytoprotection in human polymorphonuclear leucocytes. AB - An in vitro study examining the effects of zinc treatment on human PMN cell phagocytosis and killing of Staphylococcus aureus and Staphylococcus epidermidis and the cytoprotection of zinc against staphylococcal toxins. Phagocytosis was studied by transmission electron microscopy using different microbiological techniques, one of which was designed to follow the kinetics of bacterial killing. No effect was found on phagocytosis and bacterial killing. The cytotoxic effects of a crude toxin and an alpha-toxin extracted from Staphylococcus aureus preparations were studied on human PMN cells using the standard 51Cr release assay. Both toxins induced a dose-dependent leakage of 51Cr, indicating cell membrane damage. These results were confirmed by electron microscopy during the phagocytosis of S. aureus, where severe PMN cellular degeneration was observed. The addition of zinc to PMN cells strongly inhibited the release of 51Cr. In conclusion, our results show that zinc in higher than physiological concentrations does not inhibit PMN cell functions such as phagocytosis and intracellular killing of S. aureus and S. epidermidis. The addition of zinc may be beneficial in certain clinical situations, such as wound healing, zinc deficiency and infections involving toxin-producing bacteria, e.g. S. aureus. PMID- 7488385 TI - Intermediate filament expression and lectin histochemical features of canine transmissible venereal tumour. AB - Immunocharacterization of intermediate filament proteins and lectin-binding studies were carried out in canine transmissible venereal tumour (TVT), a unique neoplasia sharing some epidemiological features with Kaposi's sarcoma in humans. Neoplastic cells express vimentin, but neither cytokeratin nor desmin. Regarding lectins, TVT cells express receptors for Triticum vulgaris (WGA), Concanavalia ensiformis (Con A) and Ricinus communis I (RCA-I). They appear to be negative for Ulex europaeus-I (UEA-I), Arachis hypogaea (PNA), Glycine maximus (SBA) and Dolichos biflorus (DBA). PMID- 7488386 TI - Proposed new bacterial taxa and proposed changes of bacterial names published during 1994 and considered to be of interest to medical and veterinary bacteriology. AB - At the recent meeting of the International Committee on Systematic Bacteriology (ICSB) in 1994 it was decided to establish an ad hoc committee within the ICSB with the task of formulating an approach to communication of new names to users in medical and veterinary bacteriology. This is the first list which includes 61 names. PMID- 7488387 TI - Bcl-2 oncoprotein is widespread in lymphoid tissue and lymphomas but its differential expression in benign versus malignant follicles and monocytoid B cell proliferations is of diagnostic value. AB - The distribution of Bcl-2 oncoprotein was studied immunohistochemically in formaldehyde-fixed and paraffin-embedded reactive and neoplastic lymphoid tissue. The potential of Bcl-2 for the differential diagnosis of follicular lesions was emphasized, and the results on follicular lesions were correlated with those of polymerase chain reaction (PCR) assay of the immunoglobulin heavy chain gene rearrangement. In hyperplastic lymphoid tissue, Bcl-2 reactivity was widespread, including germinal center surroundings, scattered cells within the germinal centers, and the T-cell areas in general. Distinctively negative lymphoid populations included the majority of germinal center cells, and the negative staining pattern was maintained in cases of florid hyperplasia. In contrast, follicular lymphoma cells were consistently Bcl-2 positive. The immunohistochemical Bcl-2 reactivity of lymphoma follicles correlated with the clonal PCR amplification pattern of the immunoglobulin heavy chain gene; all Bcl 2-negative hyperplasias revealed a non-clonal pattern. Clusters of monocytoid B cells were Bcl-2 negative, whereas monocytoid B-cell lymphomas and closely related MALT lymphomas were positive. All other small cell non-Hodgkin's lymphomas of B-cell types showed nearly uniform Bcl-2 reactivity, whereas large cell B-cell lymphomas were variably positive (74%). In Hodgkin's cells, Bcl-2 reactivity was seen in the neoplastic populations of most cases of nodular sclerosis and mixed cellularity types, whereas the L&H and Reed-Sternberg cells in lymphocyte predominance Hodgkin's disease were negative in most cases. Bcl-2 immunohistochemistry thus appears very valuable in the differential diagnosis of follicular hyperplasia and neoplasia, and it may help to distinguish between reactive and neoplastic monocytoid B cells. However, Bcl-2 immunohistochemistry is not useful in the subtyping of B-cell lymphomas. PMID- 7488389 TI - Thymomas and thymic carcinomas. A retrospective investigation with histological reclassification. AB - The morphological heterogeneity of thymomas has caused much confusion respecting their classification. Recently Kirchner & Muller-Hermelink (4) proposed a histological subclassification which has been claimed to represent an independent prognostic factor: medullary and mixed thymomas are benign; organoid and cortical type as well as well-differentiated thymic carcinomas are low-grade malignant tumors, which have the capacity to recur and spread, even if they are clinically benign. High-grade malignant thymomas are always malignant. We present the clinicopathological data on 10 clinically benign and 14 clinically malignant thymomas. Having reclassified the thymomas, we found four low-grade malignant examples among the clinically benign thymomas. It is important to identify this group of patients as they are at risk of tumor recurrence. However, further investigation is needed to support the validity of this subclassification system. PMID- 7488388 TI - Folate receptor in malignant effusions of ovarian carcinoma. AB - Binding of 3H-folate in human ovarian adenocarcinoma tissue was of a high affinity type (K approximately 10(10) M-1) and displayed apparent positive cooperatively. A high-affinity folate receptor was also present in ascitic fluid and pleural effusion. Radioligand dissociation was slow at pH 7.4, but rapid at pH 3.5. The folate analogues methotrexate and in particular 5 formyltetrahydrofolate acted as inhibitors of 3H-folate binding in ascitic fluid. Ovarian adenocarcinoma tissue showed immunostaining with rabbit antibodies against human milk folate-binding protein. The gel filtration diagram contained two peaks of radiolabelled folate (at 25 and 100 kDa). The 25 kDa peak was predominant in ascitic fluid and pleural effusion. A single band of 70 kDa was seen on SDS-PAGE immunoblotting of tissue and malignant effusions. The concentration of folate receptor in tissue and fluid specimens could be determined by an immunochemical method (ELISA) utilizing antibodies against human milk folate-binding protein. PMID- 7488390 TI - Typing of coagulase-negative staphylococci from peritonitis in CAPD-patients by the PhP-CS system and REA. AB - Coagulase-negative staphylococci (CNS) were the most common bacteria causing peritonitis in patients treated with continuous ambulatory peritoneal dialysis (CAPD). In order to investigate if the same clone was responsible for the peritonitis in the different patients and if the exit site was the source of infection we followed 68 patients on CAPD for 2 years. During this period 9 patients had 12 episodes of peritonitis caused by CNS. Cultures were taken from exit site and peritoneal fluid in all patients at peritonitis and during the first study year at monthly intervals. In each culture up to 10 isolates of CNS were randomly collected and frozen. All 437 CNS isolates from the patients with CNS peritonitis were typed using a biochemical typing method and 41 isolates identical by this method were further discriminated by a DNA fingerprinting method. Identical strains were in no case isolated from different patients, indicating that no virulent strain was spread between the patients. The isolates causing the peritonitis were never found at the exist sites before the first day of the peritonitis in any patient. In only two patients was the same strain found at the exit site and in the peritoneal fluid on the first day of peritonitis. It thus seems that no virulent clone of CNS was infecting the patients and we found no evidence of CNS at the exit site causing the peritonitis. PMID- 7488391 TI - Survey of extraintestinal nontyphoid Salmonella infections in a Danish region. Inverse relation of invasiveness to frequency of isolation. AB - A total of 790 cases of nontyphoid salmonellosis distributed among 44 serotypes was registered in the County of Northern Jutland, Denmark (population 487,000) during a 3-year period from January 1992 to December 1994. There were 45 cases of extraintestinal infection, 30 of which had a positive blood culture, 12 a positive urine culture, and 5 a positive culture of either pus or tissue. For the 9 serotypes represented by > or = 5 isolates the proportion of extraintestinal isolates was inversely related to overall frequency of isolation (chi-square test, f = 1, p = 0.00003). Although limited, our study is consistent with surveys in the UK and US pointing to a modifying effect of exposure on the invasiveness of nontyphoid Salmonella serotypes. PMID- 7488392 TI - Regulation of cellular retinoic acid binding protein expression in rhino mouse skin by all-trans-retinoic acid. AB - Cellular retinoic acid binding proteins (CRABP) are cytoplasmic proteins that bind all-trans-retinoic acid (RA) and other retinoids. The purpose of these studies was to determine the effects of topically applied RA on CRABP expression in rhino mouse skin. CRABP-II mRNA was significantly induced (3- to 4.5-fold) by a single dose of RA at 6 and 16 h after RA treatment, with a return to control levels at 48 h. CRABP-II message was not significantly elevated by 3 or 4 consecutive days of RA treatment, when assessed 24 h after the last treatment. CRABP-I mRNA was undetectable in control and RA-treated skin. We used radiolabelled RA binding combined with non-denaturing PAGE blot autoradiography to distinguish the CRABP subtypes. By this protein assay method, increases in CRABP-II were detected 24 and 48 h after a single application of RA, as well as after 3 and 4 days of RA treatment. RA treatment did not alter CRABP-I expression relative to the vehicle control. These results demonstrate that in mouse skin CRABP-II, but not CRAB-I, is inducible by RA, and is similar to how RA regulates CRABP in human skin. PMID- 7488393 TI - Distribution and keratolytic effect of salicylic acid and urea in human skin. AB - Salicylic acid (SA) and urea are widely used in topical preparations. Using a simple tape stripping technique the effect on the binding forces within the stratum corneum and the skin absorption of SA and urea were studied. The degree of stratum corneum removal was recorded by measuring the transmission through the tape with a digital light-measuring instrument. With successive stripping of the skin the amount of tissue adhering to the tape decreased. Exposure of the upper arm to 2% SA for 6 h increased the skin material on the tape strips significantly. No significant increase was recorded after 3-hour exposure, or after exposure to 0.5% SA. Neither did the exposure to 10% urea for 3 or 6 h influence the amount of skin adhering to the tape significantly. Radiochemical analyses showed that the amount of SA and urea in each of the first 6 tape strips was about 5-15 micrograms/cm2. This technique provides a useful tool to evaluate the binding forces within the stratum corneum in relation to absorption of topically applied substances. PMID- 7488394 TI - Use of continuous electrical capacitance and transepidermal water loss measurements for assessing barrier function in neonatal rat skin. AB - A vital function of the skin is to oppose the loss of water to the environment. In this study, two complementary methods, transepidermal water loss (TEWL) and continuous electrical capacitance under occlusion, were used to assess epidermal barrier function in a developmental animal model, the neonatal Sprague-Dawley rat. TEWL monitors barrier function directly while the increase in capacitance under occlusion is related to both the skin's barrier function and to its water holding capacity. Serial tape stripping of the stratum corneum on 1-day-old rat pups led to a significant increase in both TEWL and continuous capacitance measurements. Anatomic site heterogeneity and the effects of surface temperature were also studied. The ventral skin surface exhibited an increase in the continuous capacitance measurements, an effect possibly due to the thinner stratum corneum on the ventral side. Both TEWL and continuous capacitance values were directly correlated with ambient temperatures within the physiological range. PMID- 7488395 TI - Study of the stratum corneum barrier function by transepidermal water loss measurements: comparison between two commercial instruments: Evaporimeter and Tewameter. AB - The measurement of transepidermal water loss (TEWL) is an important noninvasive method in order to assess the barrier function of the stratum corneum. As a consequence, TEWL has been found to be a very useful technique for studying skin irritation induced by various physical and chemical effects. Exposure of the skin to chemicals (detergents) and physical conditions (occlusion and stripping) generally results in an increase of TEWL. Until recently the only commercially available TEWL instrument was the Evaporimeter developed by ServoMed (Sweden). The instrument is based on the open chamber system with two humidity and temperature sensors which measure the water evaporation gradient at the surface of the skin. Recently a new commercially available instrument based on the same physical principle of measurement was developed by Courage and Khazaka (Tewameter, FRG). Our aim in this study has been to compare the measuring capacities of both instruments. The accuracy, sensitivity, variability and reproducibility of both instruments were compared in vivo under identical conditions on normal skin and skin damaged by external conditions. The influence of external and environmental factors such as air and probe temperature, relative humidity, air turbulence and pressure of application was evaluated for both instruments. Finally, the two instruments were compared in practice when following the increase in TEWL of the skin after stripping, occlusion and exposure to irritant household detergents. A very good correlation (r = +0.97) was found between the results of the two instruments over a wide range of TEWL values. PMID- 7488396 TI - Dapsone inhibits the generation of 5-lipoxygenase products in human polymorphonuclear leukocytes. AB - Dapsone (4,4'-diaminodiphenylsulfone) is effective in treating inflammatory skin diseases. Several lines of evidence suggest that the anti-inflammatory properties of this sulfone are partially due to modulation of functions of polymorphonuclear leukocytes (PMN). The goal of the present investigation is therefore to ascertain possible inhibitory effects of dapsone upon human 5-lipoxygenase (5-LOX) pathway. PMN of healthy donors were pretreated with dapsone in different concentrations (1.6-100 microM) for 5 min following by adding Ca ionophore A 23187 (10 microM) and subsequent incubation for 10 min. Thereupon the eicosanoids were assessed by reversed-phase high-performance liquid chromatography (RP-HPLC). Dapsone exhibited dose-dependent inhibitory activity showing 50% inhibition at 15 microM for leukotriene B4 (LTB4) with 5 x 10(6) PMN. The IC50 (half maximum inhibition concentration) of dapsone for 5-hydroxyeicosatetraenoic acid (5-HETE) and omega OH-LTB4 amounted to similar values (5-HETE: 9 microM; omega-OH-LTB4: 11 microM). The inhibition of the conversion of arachidonic acid to several eicosanoids mainly suggests an effect on the 5-LOX enzyme. The comparison of inhibition values between intact PMN and a cell-free system (by sonification) indicates an additional effect of dapsone upon enzymes other than 5-LOX. Since the concentrations used are comparable with therapeutic conditions, dapsone may exert part of its anti-inflammatory effect by prevention of the generation of 5-LOX metabolites. PMID- 7488397 TI - Modification of lipogenesis in the isolated hamster flank organ through clofibric acid. AB - The hamster flank organ model was used to measure the effects of clofibric acid on the in vitro uptake of sodium [1-14C] acetate into lipids. Clofibric acid produces a dose-dependent inhibition of lipogenesis in the hamster flank organ in vitro. PMID- 7488398 TI - Skin thickness in patients with osteoporosis and controls quantified by ultrasound A scan. AB - In order to study a possible association between skin thickness and osteoporosis, we measured skin thickness by A mode ultrasound scanning in 20 females with osteoporosis and 20 age- and sex-matched controls. Bone mineral content (BMC) of the lumbar spine and the forearm was measured by dual-photon absorptiometry. BMC of the lumbar spine was significantly reduced in the osteoporotic group as compared to controls (p < 0.002). No difference in skin thickness was found between osteoporotic patients and controls. A statistically significant correlation between skin thickness on the forearm and BMC of the forearm was found (p < 0.02), but was opposed by a lack of correlation between skin thickness and BMC of the lumbar spine. We found a correlation between skin thickness and body weight (p < 0.002), which to our knowledge had not been reported earlier. PMID- 7488399 TI - Anti-inflammatory activity of croconazole, a broad-spectrum antimycotic agent, in the arachidonic acid-induced mouse ear swelling test. AB - Investigations on croconazole, a novel imidazole compound, suggested antiphlogistic properties in vitro. Hence, its anti-inflammatory capacity was tested in vivo using the arachidonic acid-induced mouse ear swelling test, which is a suitable model for screening inhibitors of the lipoxygenase and/or the cyclooxygenase. Topical application of croconazole (1%/0.01%) to the mouse ear induced a maximal inhibition of edema (inhibition: 39%/33%; p = 0.01) which was as strong as the reference nordihydroguaiaretic acid (inhibition: 38.9%; p = 0.01). These results justify further investigations on croconazole to study potential inhibitory effects on proinflammatory arachidonic acid metabolites. PMID- 7488400 TI - Why can't we cure primary liver cancer? PMID- 7488401 TI - Prostate cancer--what should be studied? PMID- 7488402 TI - Lumpectomy versus quadrantectomy for breast conservation: a critical appraisal. PMID- 7488403 TI - Routine or delayed axillary dissection for primary breast cancer? AB - Prophylactic lymph node excision has long been recommended for preventing axillary recurrence of primary breast cancer, and has more recently gained support from the finding that adjuvant systemic therapy preferentially benefits patients with axillary node metastases. Despite these justifications, medical opinion in many communities has become deeply polarised over the merits of routine axillary dissection. A factor likely to be contributing to this split is the popularity of prescribing adjuvant systemic therapy (usually tamoxifen) on an expectant basis. Since there has been no controlled assessment of the net benefits of axillary dissection in patients receiving routine adjuvant systemic therapy--followed where necessary by delayed ("salvage") axillary treatment- objective data are urgently needed. If no substantial benefit is lost by replacing routine with delayed dissection, a small but significant improvement in quality of life could be expected for the majority of breast cancer patients. PMID- 7488405 TI - Eicosanoids and radiation. PMID- 7488406 TI - Haemopoietic stem cell inhibition: potential for dose intensification. PMID- 7488404 TI - Breast conservation is a safe method in patients with small cancer of the breast. Long-term results of three randomised trials on 1,973 patients. AB - Breast conservation has become well-established in the treatment of early mammary carcinoma. However, a standardised treatment modality has not emerged. We have analysed the data from 1,973 patients treated in three consecutive randomised trials by four different radiosurgical procedures: Halsted mastectomy, quadrantectomy plus radiotherapy, lumpectomy plus radiotherapy, and quadrantectomy without radiotherapy, to compare the outcomes of these procedures in terms of local recurrence rate and overall survival. Eligibility criteria were similar in the three trials, and comparability between the four subgroups was excellent. Median follow-up for all patients was 82 months. The annual rates of local recurrence varied markedly according to the treatment. Patients treated with Halsted mastectomy and quadrantectomy plus radiotherapy had low annual rates of local recurrence (0.20 and 0.46, respectively) while both lumpectomy plus radiotherapy and quadrantectomy without radiotherapy had significantly higher rates (2.45 and 3.28, respectively). Patients under 45 years of age had a much higher incidence of local recurrences, while in women over 55 years local recurrences were much less frequent. Overall survival curves were identical in the four groups of patients, so that the three breast conserving radiosurgical procedures had the same survival rates as Halsted mastectomy. However, local recurrence rates were markedly influenced by the treatment method, patient age and specific histological features. PMID- 7488407 TI - Epirubicin, cisplatin and infusional 5-fluorouracil (5-FU) (ECF) in hepatobiliary tumours. AB - Hepatobiliary tumours are rare, often present late and have a poor prognosis, with no current effective systemic therapy available. This study aimed to evaluate the activity and toxicity of epirubicin, cisplatin and continuous infusional 5-fluorouracil (5-FU) (ECF) in patients with these tumours. From March 1991 to November 1993, 25 patients with advanced biliary tumours and 7 with hepatoma were treated with epirubicin 50 mg/m2 and cisplatin 60 mg/m2 intravenously (i.v.) day 1, each given every 21 days and 5-FU 200 mg/m2/day given as a continuous 24 h i.v. infusion throughout the treatment course. 8 of the 20 (40%) evaluable patients with biliary tumours responded. Median duration of response was 10 months. 2 of the 7 (29%) patients with hepatoma responded. The regimen was well tolerated with minimal haematological and non-haematological toxicity. This novel regimen is active in advanced hepatobiliary tumours. PMID- 7488408 TI - Clinical stage B non-seminomatous germ cell testis cancer: the Indiana University experience (1965-1989) using routine primary retroperitoneal lymph node dissection. AB - Between 1965 and 1989, 1180 patients at Indiana University, U.S.A., underwent retroperitoneal lymph node dissection (RPLND) for non-seminomatous germ cell (NSGC) testis cancer of whom 638 cases had primary RPLND. A subset of 174 cases were considered clinical stage B (or II) before surgery (retroperitoneal nodal metastases by clinical staging). Surgery revealed that 23% (n = 41) had pathological stage A disease (no cancerous nodes). This error rate in clinical staging has decreased somewhat with improved techniques, but remains approximately 20% over the last decade. The relapse rate in pathological stage A (n = 41) was 5% (n = 2), both of whom were cured by chemotherapy. The relapse rate in pathological stage B without postoperative adjuvant treatment (n = 54) was 35% (n = 19); 2 patients died. This indicates that 65% of pathological stage B cases were cured by RPLND alone. From 1979 to 1989, the 140 pathological stage B cases participated in a randomised prospective trial of post-RPLND adjuvant chemotherapy versus no postoperative treatment. Forty two per cent (n = 59) received postoperative platinum-based therapy (two cycles), and there has been no relapse after RPLND for stage B disease. While advances in chemotherapy for NSGC testis cancer have led to its application by several study groups to clinical stage B (or II) testis cancer (with surgery reserved only for those in partial remission), the equivalent cure rate with RPLND surgery with chemotherapy rescue reserved for those who relapse appears to have both cost and risk-benefit advantages. PMID- 7488409 TI - Assessment of five serum marker assays in patients with advanced breast cancer treated with medroxyprogesterone acetate. AB - This study concerns five different tumour marker assays examined in the context of 94 patients with advanced breast cancer treated in a prospectively randomised trial of different doses of medroxyprogesterone acetate (MPA). MPA was administered at doses of 500 or 1000 mg daily and clinical evaluation of patients was carried out according to UICC criteria. Carcinoembryonic antigen (CEA) was selected as a standard marker, with three assays for MUC1 mucins (epithelial mucin core antigens (EMCA), EMCA2 and BR-MA immunoradiometric assay) differing in antibody specificities for different mucin epitopes. An additional novel assay for soluble cytokeratin was also evaluated as an example of an independent marker with a different nature and biology. Sensitivity of individual assays ranged between 44 (EMCA2) and 69% (cytokeratin) and the use of two assays in combination led to sensitivities as high as 84% (cytokeratin+BR-MA). The proportion of patients found to be assessable by each assay ranged between 51 (EMCA2) and 76% (cytokeratin). Of those patients whose marker changes were assessable, those receiving the higher dose of MPA displayed significant falls in marker levels after 12 weeks of treatment. This effect was not observed in patients receiving 500 mg. The change in cytokeratin levels in patients undergoing high dose MPA therapy proved to be most marked. Using the cytokeratin assay, 91% (of 23 patients) of patients with progressive disease showed at least a 25% rise in serum marker levels. Of these, 66% showed increases before disease progression was detected clinically with a mean lead time of 14 weeks. There was very little difference between the responses of the five tumour marker assays in patients with stable or responding disease, the proportion of these patients with stable or falling tumour marker levels ranging between 58% (CEA) and 77% (EMCA). We conclude that the cytokeratin assay has an application in monitoring response to therapy and predicting tumour progression in advanced breast cancer patients with assessable tumour marker profiles, especially if used in combination with a MUC1 mucin assay. PMID- 7488410 TI - Intrapatient comparison of single-agent epirubicin with or without lonidamine in metastatic breast cancer. AB - The aim of this study was to determine if lonidamine (LND) supplementation to single-agent epirubicin (EPI) could reverse anthracycline resistance in patients with metastatic breast cancer. 45 patients with metastatic breast cancer were treated with EPI 120 mg/m2 by intravenous (i.v.) bolus every 3 weeks. Patients who progressed were given the same chemotherapy regimen on day 4 in combination with oral LND, 150 mg on day 1, 300 mg on day 2 and 450 mg on days 3-5. Among the 40 evaluable patients, 6 complete responses (CR) and 14 partial responses (PR) were achieved with EPI treatment alone for an overall response rate of 50%. The median duration of response was 6.5 months. Among the 25 patients treated with EPI+LND, 5 PR (21% of 24 evaluable patients) were observed with a median duration of response of 7 months. The median survival in patients receiving both treatments was 20 months. The survival for all patients was 18 months. The survival of patients receiving LND was not significantly longer than for the other patients. Myelotoxicity was the most common side-effect followed by alopecia, nausea and vomiting, and stomatitis. LND-related toxic effects were mild-to-moderate epigastralgia and myalgia. Anthracycline-related toxicity was the same in the two treatment groups. This study indicates that LND may circumvent clinical resistance to EPI without altering the pattern or severity of the toxicity of this anthracycline. Continued investigation of the clinical modulation of EPI resistance by LND in breast cancer is warranted, hopefully in patients with known multidrug resistance status. PMID- 7488411 TI - CEA, MCA, CA 15.3 and CA 549 and their combinations in expressing and monitoring metastatic breast cancer: a prospective comparative study. AB - Serum levels of carcinoembryonic antigen (CEA), mucin-like carcinoma-associated antigen (MCA), CA 15.3 and CA 549 were concurrently assayed in patients with metastatic breast cancer. Overall sensitivity in detecting metastatic breast cancer (201 pts) was CEA 45%, MCA 59%, CA 15.3 71% and CA 549 72% (P < 0.01). Sensitivity increased by only 6% to 8% when two or more antigens were simultaneously considered. An overall sensitivity of correlation with objective response (n = 71) was observed in the range of 53-67% (P = n.s.) in patients with abnormal baseline marker values, and in the range of 42-87% (P < 0.05) in patients with normal baseline values. The combination of two or more markers did not improve sensitivity, but decreased specificity of correlation with objective response. In conclusion, CA 15.3 and CA 549 have individually higher sensitivity in detecting metastatic breast cancer. No clinical advantage was observed for using two or more markers concurrently over CA 15.3 or CA 549 alone in the monitoring of metastatic breast cancer. PMID- 7488412 TI - High dose epirubicin is effective in measurable metastatic prostate cancer: a phase II study of the EORTC Genitourinary Group. AB - 39 hormone-resistant prostate cancer patients with bidimensionally measurable metastatic lesions were given epirubicin 100 mg/m2 intravenously every 3 weeks. One patient was ineligible and excluded from analyses. According to WHO criteria, 9 patients (24%) had a partial response, 16 patients (42%) had stable disease (including 3 patients (8%) with a partial response not confirmed 1 month later), 11 patients (29%) had progressive disease, and in 2 patients (5%) response was not evaluated. Toxicity was as expected. Fifty-five per cent of patients had WHO grade 3/4 toxicity for white blood cells, and 3% of patients grade 3 toxicity for platelets. Other toxicities included nausea and vomiting, mucositis and alopecia. 2 patients with pre-existing cardiac disease developed cardiotoxicity (1 grade 2, 1 grade 3). An attempt was made to correlate response with prostate specific antigen (PSA) measurements. A positive trend was seen, but 2 non-responding patients showed a > 50% decrease in value. PMID- 7488413 TI - Immunocytochemical detection of isolated tumour cells in bone marrow of patients with untreated stage C prostatic cancer. AB - The micrometastatic spread of tumour cells is usually missed by conventional diagnostic techniques, although this spread largely determines the prognosis of patients with primary epithelial cancers. By use of the monoclonal antibody, CK2, to epithelial cytokeratin component number 18 (CK18), individual disseminated carcinoma cells present in bone marrow of cancer patients can now be identified. In the present study, this approach has been applied to patients with virginal stage C adenocarcinoma of the prostate. Double-sided aspirates of iliac bone marrow from 24 of 44 evaluable patients (54.4%) exhibited between one and 38 CK18 positive cells per sample of 2 x 10(6) mononuclear cells. In 13 of these 24 positive patients, CK-positive cells were only detected in one of the two aspirates analysed. There was no statistically significant correlation between this finding and established risk factors, such as the volume and histological grade of the primary tumour or the concentration of prostate specific antigen and prostatic acid phosphatase in serum. The follow-up time is too short to provide meaningful data on the prognostic significance of isolated CK18-positive cells in bone marrow, which, however, has been recently demonstrated in other types of primary epithelial cancers. In conclusion, the presence of prostatic tumour cells in bone marrow might be interpreted as an indicator of the metastatic capacity of an individual primary tumour. The immunocytochemical detection of these cells may, therefore, be useful for increasing the precision of current tumour staging, and to monitor minimal residual cancer in an individual patient. PMID- 7488414 TI - Accelerated cisplatin-based chemotherapy for advanced bladder cancer. AB - The toxicity and efficacy of accelerated cisplatin, vincristine and methotrexate was assessed in patients with advanced urothelial carcinoma. 30 consecutive patients were entered into a phase II trial and treated with cisplatin, vincristine and methotrexate given every 10 days (MOPq10) for four cycles, followed by two further cycles at 21 day intervals. Five complete responses and 14 partial responses were observed (overall response rate 63%; 95% confidence interval 45-78%). The median progression-free survival was 7.5 months (range 1.8 28) and the median overall survival 10.5 months (range 2.36). Toxicity was moderate with no treatment-related deaths. It is concluded that although the overall survival is still disappointing, the toxicity is less with the protocol than reported with methotrexate, vinblastine, doxorubicin and cisplatin (M-VAC) or escalated M-VAC (E-MVAC) and the time on treatment is shorter. MOPq10 provided palliative benefit to two-thirds of patients with advanced transitional cell carcinoma including those in their eighth decade. PMID- 7488415 TI - A prospective randomised study in limited disease small cell carcinoma- doxorubicin and vincristine plus either cyclophosphamide or etoposide. AB - A prospective randomised study was undertaken in patients with limited disease small cell carcinoma of the lung (SCCL), which compared doxorubicin, 50 mg/m2, and vincristine, 2 mg i.v. (intravenously) on day 1, with either cyclophosphamide, 800 mg/m2 on day 1 (CAV) or etoposide, 60 mg/m2 i.v. on day 1 and 120 mg/m2 orally on days 2-5 (AVE). Responding patients were to receive six cycles of chemotherapy at 3 weekly intervals followed after 2 weeks by mediastinal irradiation. Response rates and toxicity were evaluated by the chi square or Fisher's exact test and survival by the logrank test. 81 patients were entered into the study, 38 of whom received CAV and 43 received AVE. In the patients treated with CAV and AVE, the overall response rate was 61% (confidence limit (CL), 45-71%) and 74% (CL, 61-87%) respectively, the complete response rate was 32% (CL, 17-47%) and 51% (CL, 36-66%), respectively (P = 0.07) and the median survival was 12 and 14.5 months, respectively (P = 0.15). In the patients treated with CAV and AVE, the incidence of grade 3 and 4 leucopenia was 29% (CL, 15-43%) and 9% (CL, 0-18%), respectively (P = 0.025). No patient developed doxorubicin cardiomyopathy. These findings support the role of etoposide in first line chemotherapy for SCCL. AVE is among the more efficacious regimens for SCCL and also has a relatively low toxicity. PMID- 7488416 TI - A cost analysis of Nd:YAG laser ablation versus endoscopic intubation for the palliation of malignant dysphagia. AB - Although endoscopic intubation is the mainstay of non-surgical palliation of malignant dysphagia, Nd:YAG laser ablation has been shown to provide good palliation with few complications. The study reported here incorporates data from published and unpublished sources into a cost model which estimates the lifetime cost of palliation with the two therapies. It is estimated that, depending on the assumptions used, laser palliation costs between 153 pounds and 710 pounds more per patient than endoscopic intubation. Sensitivity analysis is used to assess whether variation in clinical practice and in the unit costs of resources will change the conclusions of the study. This indicates that, under most alternative sets of assumptions, intubation retains its cost advantage. However, factors that might reduce, or even eliminate, this cost differential include undertaking more laser procedures as day-cases, using more expensive expanding metal stents for intubation and reducing the need for follow-up laser procedures with palliative radiotherapy. PMID- 7488417 TI - Comparison of granisetron alone and granisetron plus hydroxyzine hydrochloride for prophylactic treatment of emesis induced by cisplatin chemotherapy. AB - The efficacy and safety of granisetron alone (group G) and granisetron plus hydroxyzine hydrochloride (group G/H) as prophylactic therapy for acute and delayed nausea and vomiting were evaluated in an open trial in head and neck cancer patients undergoing chemotherapy with cisplatin. The severity of nausea was significantly reduced on days 1 and 4 in patients receiving combination therapy, but the frequency of vomiting was not significantly different between the two groups. The only side-effect observed was headache in 1 patient from group G, and no drug-related laboratory test abnormalities were observed. These results suggest that the anti-emetic efficacy of granisetron can be augmented by hydroxyzine hydrochloride. PMID- 7488419 TI - Adult cancer risks among the mothers of children with cancer. AB - We investigated cancer risk among mothers of 2365 children who were diagnosed with cancer between 1973 and 1989 in Sweden. From the date of birth of the child until 31 December 1989, 38 cases of cancer were diagnosed among the mothers. The expected number of cases, according to national rates, was 30.9. Cancer of the thyroid was the only site showing a significantly increased risk among the mothers (observed = 6, expected = 1.2, P < 0.01), but two cases were medullary cancer associated with an inheritable syndrome, and inherited by their children. PMID- 7488418 TI - Evaluation of the informed consent procedure in cancer patients candidate to immunotherapy. AB - By addressing questionnaires to 24 cancer patients candidate to immunotherapy, we evaluated both the effects and the effectiveness of the medical information provided to the patient on their knowledge of the disease and the treatment. Most patients had correctly understood the information but 69% stated that they had been unable to ask all the questions they wished, and 62% required additional information. Most patients admitted to being emotionally distressed throughout the interview. These results are not significantly different from those obtained in patients candidate to new chemotherapy agents, but show that important improvements in the informed consent procedure are required. PMID- 7488420 TI - Risk analysis in acute lymphoblastic leukaemia: problems and pitfalls. PMID- 7488422 TI - Diverging trends in colorectal cancer morbidity and mortality. Earlier diagnosis comes at a price. AB - In developed countries, time trends in the incidence of colorectal cancer differ markedly from trends in mortality. This study sought to explain simultaneously changes in both colorectal cancer incidence and mortality. Data on first admissions, interventions and outcome from the national hospital registry over the period 1978-1989 and data on mortality from Statistics Netherlands over the same period were analysed by age-period models and subsequently entered in a Markov chain model, simulating disease history from first admission to death. Over the period 1978-1989, age adjusted numbers of first admissions and interventions increased by 37% and 32%, respectively, while mortality declined by 8%. For every 100 patients admitted between 1987 and 1989, 13 more will survive compared with 1978-1980. Of these, 3 will be saved by improving results of primary treatment but the other 10 will survive their diagnosis for the subsequent 10 years. Although progress in treatment has been made, therapeutic improvement can account only for the smaller part of the divergence between morbidity and mortality. Increased diagnostic activity, raising incidence and lowering mortality simultaneously, is the most likely cause of the unexplained divergence. PMID- 7488423 TI - The management of cervical intra-epithelial neoplasia (CIN): extensiveness and costs in The Netherlands. AB - In order to provide greater insight into both the extensiveness and the medical costs of the diagnosis and treatment of screen-detected cervical intra-epithelial neoplasia (CIN) in general medical practice in The Netherlands, data from national registries and gynaecology departments were retrieved, and experts were interviewed. Of the 5060 women diagnosed with CIN in 1988, more than 50% were treated in hospital with conisation or hysterectomy, which on average took 5.5 days stay per admission. The assessed average duration of the total pre- and post treatment period is 4.6 years. The average total medical costs in women with detected CIN III are Dfl 3700 per woman. The diagnosis of CIN I and II involves more medical procedures and time than CIN III, but fewer women have conisation or hysterectomy, resulting in lower total medical costs (Dfl 2572). The overall extent and costs of the management of CIN should be accounted for when balancing the benefits, unfavourable effects and costs of cervical cancer screening. PMID- 7488421 TI - Socioeconomic status and breast cancer survival in the southeastern Netherlands, 1980-1989. AB - Socioeconomic differences in breast cancer survival in the southeastern Netherlands between 1980 and 1989 were studied (n = 3928), as was the impact of prognostic factors (stage at diagnosis, morphology, and treatment) on such differences. An area-based measure of socioeconomic status (SES) in five groups, based on the postcode of residence at the time of diagnosis, was used. In univariate analyses the relative survival rate was used to correct for causes of death other than breast cancer. The measure of outcome in multivariate analyses was the hazard ratio. The results of both univariate and multivariate analyses suggested a small survival advantage for the higher SES groups. In a model with follow-up period, SES and age, the hazard ratios with 95% confidence intervals (CI) for SES groups from high to low were: 1.00, 1.06 (0.84-1.33), 1.04 (0.86 1.26), 1.15 (0.96-1.38), 1.18 (0.99-1.42). After a correction for stage at diagnosis, differences in survival were reduced substantially. Morphology and treatment were not important explanatory factors of the SES survival association. We conclude that small socioeconomic differences in breast cancer survival exist in The Netherlands and that stage at diagnosis is the most important determinant of such differences. PMID- 7488424 TI - Activity of NK 611, a new epipodophyllotoxin derivative, against colony forming units from freshly explanted human tumours in vitro. AB - NK 611 is a new semisynthetic analogue of etoposide, which presumably also acts through inhibition of topoisomerase II, and has been found to be more potent against several cancer cell lines in vitro than etoposide. The objectives of our study were to determine the activity of NK 611 against freshly explanted clonogenic cells from human tumours and compare this agent with etoposide and other clinically useful agents. After exposure for 1 h in 45 evaluable tumour specimens, NK 611 showed clear concentration-dependent antitumour activity. At 51 microM, 49% of specimens were markedly inhibited. Using a long-term (21-28 day) exposure at 6.8 microM, 58% of 50 evaluable specimens were profoundly inhibited. At equimolar concentrations, NK 611 was as active as etoposide. Across all tumour types studied, NK 611 was as active as vinblastine, bleomycin, doxorubicin, 5 fluorouracil, mitomycin-C and cisplatin. Our results showed cross resistance to etoposide in the majority of specimens. Activity of NK 611 was greater with long term exposure than with short-term exposure indicating schedule dependency. We conclude that NK 611 has a wide spectrum of in vitro antitumour activity. Since preliminary clinical information suggests that this drug is well tolerated at high doses, further development of this agent in Phase II trials with multiple dosing schedules is warranted. PMID- 7488425 TI - Flow cytometric functional analysis of multidrug resistance by Fluo-3: a comparison with rhodamine-123. AB - Using four cell lines including drug-sensitive K562/Parent cells, P-glycoprotein (Pgp)-mediated multidrug resistant (MDR) K562/VCR, K562/ADR and revertant K562/ADR-R cells, two fluorescent agents, Fluo-3 and rhodamine-123 (Rh-123), were compared as indicators in a functional assay of MDR. Cells were incubated with 4 microM Fluo-3 or 1 microM Rh-123 for 45 min and then the intracellular accumulation of the agent was measured using a flow cytometer. Verapamil (20 microM) or cepharanthine (biscoclaurine alkaloid, 10 microM) was added just before the fluorescent agents. Efflux patterns were also studied 60 min after incubation with or without verapamil and cepharanthine. Increased intracellular accumulation and a delayed efflux pattern of Fluo-3 by verapamil and cepharanthine were demonstrated in multidrug resistant K562/VCR and K562/ADR cells, indicating that Fluo-3 is another good indicator of MDR. However, a similar, but lower, increase in uptake and a delayed efflux pattern of Fluo-3 by verapamil and cepharanthine were also demonstrated even in Pgp-non-overexpressed K562/Parent cells. In contrast, accumulation of Rh-123 was not affected by verapamil and cepharanthine. To further study the Pgp dependency of Fluo-3, another cell line, K562/NC16 expressing minimum MDR1 mRNA, was cloned. Increased uptake and a delayed efflux pattern of Fluo-3, but not Rh-123, with verapamil or cepharanthine were again demonstrated in K562/NC16 cells, indicating that intracellular accumulation of Fluo-3 may be non-specifically influenced by verapamil and cepharanthine at very low levels of Pgp-related MDR, while the influx and efflux patterns of Rh-123 may be specifically affected by Pgp overexpression. PMID- 7488426 TI - Repeating the avidin "chase" markedly improved the biodistribution of radiolabelled biotinylated antibodies and promoted the excretion of additional background radioactivity. AB - Immunoscintigraphy using radiolabelled biotinylated monoclonal antibodies followed by infusion of avidin as a "chase" has been recently reported to improve the biodistribution for both immunoscintigraphy and radioimmunotherapy. In this study the circulating protein-bound and avidin-binding fractions of radiolabelled biotinylated antibodies were determined serially after injection of an avidin "chase", and the effect of repeating the avidin chase was also studied. Nude mice bearing KT005 human osteogenic sarcoma were injected with radiolabelled biotinylated antitumour monoclonal antibody (OST7). After injection of an avidin chase, the protein-bound and avidin-binding fractions in plasma were determined serially using the trichloroacetate method and avidin-Sepharose gel. The biodistribution of radiolabelled biotinylated OST7 was compared after single and double avidin chases with no chase. At 6 h after the first avidin chase in mice injected with radioiodinated and technetium-labelled biotinylated OST7, 67.7% and 67.8%, respectively, of the plasma radioactivity was available for binding to avidin and was cleared from the circulation. Reinjection of avidin decreased the plasma radioactivity and improved the biodistribution of the radiolabelled biotinylated antibodies. Repeating the avidin chase markedly improved the biodistribution of the radioiodine-labelled biotinylated antibody when compared with the use of a single avidin chase. This new method for radioimmunotherapy is sure to protect the critical organs from radiation injury without decreasing the therapeutic effect. PMID- 7488427 TI - Patterns of DNA methylation of the parathyroid hormone-related protein gene in human lung carcinoma. AB - Humoral hypercalcaemia of malignancy often results from production of parathyroid hormone-related protein (PTHrP) by the tumour. We have investigated whether malignancy is associated with epigenetic changes in the PTHrP gene in lung. In normal and tumour tissue, there was a general background of nonmethylation in the PTHrP gene. In the 5' region, there appeared to be increased methylation of sites upstream of the promoter, P2. The extent of methylation increased from germ line to normal tissue to tumour tissue to tumour cell line, indicating that new methylation events in this region mark neoplastic change in lung cells. PMID- 7488428 TI - The effect of different dose levels of degradable starch microspheres (Spherex) on the distribution of a cytotoxic drug after regional administration to tumour bearing rats. AB - Tumour uptake of a radiolabelled anticancer drug, tauromustine ([14C]TCNU), was investigated, using two different routes of administration in rats with colon adenocarcinomas implanted in the liver. Intra-arterial administration produced higher concentrations of the drug and/or its metabolites in tumour tissue and lower concentrations in normal tissue compared to intravenous administration. The investigation of co-injection of [14C]TCNU intra-arterially with degradable starch microspheres (DSM) indicated that a high dose of DSM (30 mg/kg) resulted in a high concentration of radioactivity in normal liver tissue, and in adjacent organs. This unfavourable pattern was not observed with the low dose of DSM (12.5 mg/kg), which produced a significant increase in tumour uptake. The results demonstrate the efficacy of partial vascular blockade, as elicited by the low dose of DSM. This regime caused [14C]TCNU to be preferentially retained in the active peripheral regions of the tumour. PMID- 7488429 TI - Prognostic value of CD44 splice variants in human stage III cervical cancer. AB - The expression of specific cell adhesion molecule CD44 isoforms (splice variants) has been shown to be associated with poor prognosis in human malignancies, such as breast cancer. We used three different variant exon sequence-specific murine monoclonal antibodies to epitopes encoded by exons v5, v6 or v7-v8 of human variant CD44, to study the expression of CD44 splice variants by immunohistochemistry in human stage III cervical cancer. We investigated 40 pretreatment punch biopsies of cervical cancer FIGO stage III. CD44 splice variants CD44v5, CD44v6 and CD44v7-8 were detected by means of immunohistochemistry in 90%, 55% and 25%, respectively. CD44 epitopes encoded by exon v5 were not correlated with prognosis. Expression of CD44 splice variants containing epitopes encoded by exon v6 were correlated with significantly poorer prognosis (Mantel test, P = 0.008). Five-year survival rates with or without CD44v6 expression were 20% versus 71%, respectively. Expression of CD44v7-8 was also correlated with significantly poorer overall survival (Mantel test, P = 0.02). Expression of CD44 splice variants containing epitopes encoded by exons v7 v8 and especially exon v6 is associated with significantly poorer prognosis in stage III cervical cancer patients. PMID- 7488430 TI - The performance of a commercial radioligand binding assay for the epidermal growth factor receptor is comparable to the EORTC standard assay. PMID- 7488431 TI - Postmastectomy radiotherapy, axillary lymph node dissection and oestrogen receptor status predict prognosis. PMID- 7488432 TI - Spreading evaluation in primitive bronchogenic carcinoma: benefit of cerebral MRI compared to CT scan. PMID- 7488433 TI - Prognostic significance of Lewis related antigen expression in stage I non-small cell lung cancer. PMID- 7488434 TI - High dose 5-fluorouracil (5-FU) and folinic acid in advanced colorectal cancer resistant to standard dose 5-FU treatment: results of a phase II study. PMID- 7488435 TI - Ribonucleotide reductase inhibition in the treatment of advanced prostate cancer: an experimental approach with hydroxyurea and gallium nitrate in 20 patients. PMID- 7488436 TI - Active specific immunotherapy with an autologous virus-modified tumour cell vaccine in human renal cell carcinoma (RCC) PMID- 7488438 TI - A randomised double-blind placebo controlled clinical trial assessing the tolerability and efficacy of glutathione as an adjuvant to escalating doses of cisplatin in the treatment of advanced ovarian cancer. PMID- 7488437 TI - Ondansetron for the control of dacarbazine-induced emesis. PMID- 7488439 TI - Persisting elevation of alpha-fetoprotein after chemotherapy for germ cell tumour -not always due to viable malignancy? PMID- 7488440 TI - Investigation in general practice of patients with suspected heart failure. How should the essential echocardiographic service be delivered? PMID- 7488441 TI - New directions in anticoagulant and antiplatelet treatment. PMID- 7488442 TI - Long-term complications of cardiac transplantation. PMID- 7488443 TI - Angiographic trials of lipid-lowering therapy: end of an era? PMID- 7488444 TI - Rescue thrombolysis: alteplase as adjuvant treatment after streptokinase in acute myocardial infarction. AB - BACKGROUND: In acute myocardial infarction patients who do not reperfuse their infarct arteries shortly after thrombolytic treatment have a high morbidity and mortality. Management of this high risk group remains problematic, especially in centres without access to interventional cardiology. Additional thrombolytic treatment may result in reperfusion and improved left ventricular function. METHODS: Failure of reperfusion was assessed non-invasively as less than 25% reduction of ST elevation in the electrocardiographic lead with maximum ST shift on a pretreatment electrocardiogram. 37 patients with acute myocardial infarction who showed electrocardiographic evidence of failed reperfusion 30 minutes after 1.5 MU streptokinase over 60 minutes were randomly allocated to receive either alteplase (tissue type plasminogen activator (rt-PA) 100 mg over three hours) (19 patients) or placebo (18 patients). 43 patients with electrocardiographic evidence of reperfusion after streptokinase acted as controls. Outcome was assessed from the Selvester Q wave score of a predischarge electrocardiogram and a nuclear gated scan for left ventricular ejection fraction 4-6 weeks after discharge. RESULTS: Among patients in whom ST segment elevation was not reduced after streptokinase, alteplase treatment resulted in a significantly smaller electrocardiographic infarct size (14% (8%) v 20% (9%), P = 0.03) and improved left ventricular ejection fraction (44 (10%) v 34% (16%), P = 0.04) compared with placebo. This benefit was confined to patients who failed fibrinogenolysis after streptokinase (fibrinogen > 1 g/l). In patients in whom ST segment elevation was reduced after streptokinase, infarct size and left ventricular ejection fraction were not significantly different from those in patients treated with additional alteplase. CONCLUSION: Patients without electrocardiographic evidence of reperfusion after streptokinase may benefit from further thrombolysis with alteplase. PMID- 7488445 TI - Thrombolytic treatment and proteinuria. AB - OBJECTIVES: To determine whether patients with acute myocardial infarction undergoing thrombolysis with streptokinase develop changes in renal function. DESIGN: Prospective assessment of renal function in 60 consecutive patients admitted with acute myocardial infarction. SETTING: Tertiary referral centre and city general hospital. PATIENTS: 60 consecutive patients with acute myocardial infarction. Thirty eight were given streptokinase and 17 tissue plasminogen activator (alteplase) and five no thrombolytic agent (non-streptokinase group). MAIN OUTCOME MEASURES: Proteinuria and creatinine clearance on admission (day 1) and on days 3 and 6; serum urea and creatinine concentrations on days 1 and 7; streptokinase IgG on days 1, 2, and 7. RESULTS: Significant proteinuria (> 0.15 g/24 h) was found in 31 (82%) of the 38 patients in the streptokinase group (mean 0.47 g/24 h (95% confidence interval 0.35 to 0.6 g/24 h)) in the 24 hours after admission compared with six (27%) out of 22 in the non-streptokinase group (mean 0.17 g/24 h (0.12 to 0.2 g/24 h); P = 0.008). In the streptokinase group this decreased to the normal range by day 3 (mean 0.15 g/24 h (0.1 to 0.22 g/24 h); P = 0.0001 v baseline). Electrophoresis of urine showed the proteinuria to be glomerular in origin. Creatinine clearance and serum creatinine and urea concentrations were similar in both groups. In the streptokinase group detectable streptokinase IgG titres were found in 28 out of 32 (87%) patients. The median titre on admission was 16 (range 0-110); it fell to 3 (range 0-80; P = 0.001) by day 2 and increased to 61 (range 0-7700; P = 0.0002 v baseline) by day 7. CONCLUSIONS: Streptokinase was associated with significant early onset proteinuria of glomerular origin. This started to resolve by day 3 and resulted in no deterioration in overall renal function. The temporal relation to the initial fall in antibody titre suggests that it could be the result of immune complex deposition in the glomeruli. PMID- 7488447 TI - Serum lipoprotein(a) concentrations are related to coronary disease progression without new myocardial infarction. AB - OBJECTIVE: To examine the association between serum lipoprotein(a) and angiographically assessed coronary artery disease progression without new myocardial infarction. PATIENTS AND DESIGN: 85 patients with coronary artery disease who underwent serial angiography with an interval of at least two years were studied. Progression of coronary artery disease was defined as an increase in diameter stenosis of 15% or more. Vessels on which angioplasty had been performed were excluded from the analysis. The patients were classified into two groups: a progression group without new myocardial infarction (n = 48) and non progression group (n = 37). Risk factors including lipoprotein(a) were evaluated to see how they were related to progression without myocardial infarction. RESULTS: There were no differences between the two groups in the following factors: age, gender, the time interval between the angiographic studies, the distribution of the analysed coronary arteries, and history of well established coronary risk factors. Univariate analysis showed that serum lipoprotein(a) (P = 0.0002), cigarette smoking between the studies (P = 0.002), serum high density lipoprotein (P = 0.003), and serum low density lipoprotein (P = 0.01) were related to progression without myocardial infarction. Multivariate analysis selected two independent factors for progression without myocardial infarction: serum lipoprotein(a) (P = 0.003) and serum high density lipoprotein (P = 0.03). CONCLUSIONS: Serum lipoprotein(a) concentrations are closely related to the progression of coronary artery disease without new myocardial infarction. Lipoprotein(a) lowering treatment may be needed to prevent disease progression in patients with coronary artery disease and high serum lipoprotein(a). PMID- 7488446 TI - Dobutamine echocardiography and thallium-201 imaging predict functional improvement after revascularisation in severe ischaemic left ventricular dysfunction. AB - OBJECTIVES: To evaluate the concordance between thallium-201 uptake and echocardiographic wall thickening, which are both indicators of potentially reversible myocardial dysfunction, in patients with chronic ischaemic left ventricular failure and to assess their relative contribution to predicting improvement in regional function after revascularisation in a subgroup. PATIENTS AND METHODS: 45 patients with chronic ischaemic left ventricular dysfunction (mean (SD) ejection fraction 25 (8)%) underwent echocardiography before and after dobutamine infusion (10 micrograms/kg/min). Of these, 22 patients underwent rest echocardiography at a mean (SD) of 9 (1) weeks after revascularisation. 201Tl imaging was performed during dobutamine echocardiography and at rest, 1, and 4 h after treatment with sublingual glyceryl trinitrate on two separate days. Potentially reversible dysfunction was thought to be present when a myocardial segment contained a Tl score of > or = 3 (ascending score 1-4), or showed improved wall thickening of a dysynergic segment during dobutamine stimulation. RESULTS: Of the 201Tl protocols, the redistribution scan 1 h after treatment with glyceryl trinitrate best demonstrated myocardial viability. Concordance between 201Tl and dobutamine induced wall thickening was 82% (kappa = 0.59) for detecting potentially reversible myocardial dysfunction before revascularisation (n = 45). Regional function improved in 18 of 22 patients after revascularisation. There were 168 dysynergic segments before intervention. The sensitivity of echocardiography and 201Tl imaging for detecting "recoverable" or viable segments after revascularisation was 87% and 92% respectively and specificity was 82% and 78% respectively (P = NS). CONCLUSIONS: Dobutamine echocardiography and 201Tl imaging may be used to predict mechanical improvement in dysynergic segments after revascularisation in patients with chronic ischaemic left ventricular dysfunction. PMID- 7488448 TI - Absence of circadian variation in the onset of acute myocardial infarction in diabetic subjects. AB - OBJECTIVES: To investigate the circadian pattern of acute myocardial infarction in non-insulin-dependent diabetic patients and to compare it with that of controls. BACKGROUND: Previous studies have shown that there is a circadian variation in the incidence of acute myocardial infarction, but there are few data on diabetic subjects. METHODS: A hospital based prospective case-control study. RESULTS: 196 diabetic patients and 196 age and sex matched controls were admitted with a diagnosis of acute myocardial infarction during the study period. IN 32 diabetic patients and 38 controls, the time of onset of myocardial infarction was unknown; in 34, 44, 42, and 44 diabetic patients the onset was in the first to fourth quarters respectively (chi 2 = 1.66, NS). The corresponding figures for the controls were 30, 56, 45, and 27 (chi 2 = 13.9, P < 0.005). The difference between the two groups was highly significant (chi 2 = 10.3, P < 0.025). CONCLUSIONS: Diabetic subjects do not show a significant circadian variation in the onset of acute myocardial infarction. PMID- 7488449 TI - Failure of "effective" treatment for heart failure to improve normal customary activity. AB - OBJECTIVES: To examine the effects of drug treatment on laboratory exercise tests in relation to measures of daily activity in patients with chronic heart failure. SETTING: University teaching hospital. SUBJECTS: 18 patients with mild to moderate chronic heart failure (New York Heart Association functional class II III) and 10 age matched healthy controls. METHODS: Assessments were made before and after 12 weeks of vasodilator drug treatment. Exercise capacity was measured during two different types of treadmill exercise, one using a ramp protocol and the other a fixed work load. Corridor walk tests at three self selected speeds were also undertaken and measures of customary activity assessed from pedometer scores. RESULTS: Exercise times were significantly increased from baseline (P < 0.01) with both treadmill protocols after 12 weeks of drug treatment, with a positive correlation between the duration of treadmill exercise for both protocols (r = 0.69, P < 0.01). Corridor walk tests of 100 m at a self selected slow speed also improved (P < 0.02) but these did not correlate with the changes in treadmill exercise time. The pedometer scores of the patients with heart failure were greatly reduced compared with those of the controls (258 (45) x 10(2) v 619 (67) x 10(2) steps/week, P < 0.001) and after 12 weeks of treatment were unchanged (261 (42) x 10(2) steps/week). CONCLUSIONS: These data confirm the need to use different exercise protocols when assessing the benefits of drug treatment in patients with chronic heart failure. Treatments that seem effective with conventional laboratory based exercise tests may not improve daily activities. This may reflect a failure of apparently successful treatment and should be considered when interpreting clinical trials. PMID- 7488450 TI - Anatomical dead space, ventilatory pattern, and exercise capacity in chronic heart failure. AB - BACKGROUND: Patients with chronic heart failure have an excessive ventilatory response to exercise, characterised by an increase in the slope of the relation between ventilation and carbon dioxide production (VE/VCO2 slope). Patients have an altered respiratory pattern with an increased respiratory rate (f) at a given tidal volume (VT), which may result in increased anatomical dead space ventilation. METHODS: The ventilatory responses in 88 patients with chronic heart failure and 43 age matched controls during maximal incremental treadmill exercise were analysed. Peak oxygen consumption (VO2), VE/VCO2 slope, and the slope of the relation between f and VT were derived. Anatomical dead space was estimated from a standard formula and anatomical dead space ventilation calculated. RESULTS: Peak VO2 was greater (mean (SD)) (33.2 (8.5) v 19.4 (6.7) ml/min/kg; P < 0.001) and the VE/VCO2 slope lower in the controls (25.96 (4.16) v 35.14 (9.80); P < 0.001). During matched submaximal exercise VT was higher (1.97 (0.92) v 1.68 (0.62) 1; P < 0.05) and flower in the controls (18.23 (6.48) v 24.28 (7.58); P < 0.001). At peak exercise there was no difference in f, but VT was higher in the controls (2.66 (0.97) v 1.90 (0.61) 1; P < 0.001). The VT/f slope was the same (0.04 (0.04)) in both groups. The intercept of the relation was greater for the control group (1.31 (1.28) v 0.59 (0.83); P < 0.001). Anatomical dead space ventilation was lower in the controls at submaximal work load (4.17 (1.56) v 5.58 (1.93) l/min; P < 0.001). At peak exercise anatomical dead space ventilation was the same in both groups, but was lower expressed as a percentage of total VE in the control group (9.8 (3.3) v 13.5 (4.0); P < 0.001). There were weak relations within the heart failure group alone between VT/f slope and peak VO2 and VE/VCO2 slope. CONCLUSIONS: The relation between anatomical dead space ventilation and VE/VCO2 slope is expected: as f increases, so do VE/VCO2 slope and anatomical dead space ventilation. The VT/f slope was the same in patients with chronic heart failure and controls, so change in respiratory pattern cannot explain the increase in VE/VCO2 slope. The stimulus causing the increased f has yet to be identified. PMID- 7488451 TI - Inspiratory muscle strength is a determinant of maximum oxygen consumption in chronic heart failure. AB - OBJECTIVE: To investigate the significance of respiratory muscle weakness in chronic heart failure and its relation both to maximum oxygen consumption during cardiopulmonary exercise testing and to skeletal muscle (quadriceps) strength. SUBJECTS: Seven healthy men aged 54.9 (SEM 4.3) years and 20 men with chronic heart failure aged 61.4 (1.6) years (P = 0.20) with radionuclide left ventricular ejection fraction of 25.4 (3.0)%. METHODS: Mouth pressures during maximum static inspiratory effort (PImax) at functional residual capacity (FRC) and residual volume (RV) were measured in all subjects and taken as indices of inspiratory muscle strength. Similarly, mouth pressures during maximum static expiratory effort (PEmax) at FRC and total lung capacity (TLC) were taken as indices of expiratory muscle strength. Cardiopulmonary exercise testing was performed in all subjects. All controls and 15 heart failure patients also had their right quadriceps muscle strength measured. RESULTS: There was respiratory muscle weakness in heart failure patients, with reduction of PImax at FRC (59.7) (6.3) v 85.6 (9.6) cm H2O, P = 0.045), PEmax at FRC (94.8 (6.2) v 134.6 (9.1) cm H2O, P = 0.004), and PEmax at TLC (121.7 (8.5) v 160.7 (13) cm H2O, P = 0.028). PImax at RV was also reduced but this did not reach statistical significance (77.3 (6.6) v 89.3 (13) cm H2O, P = 0.44). There was also significant weakness of the right quadriceps muscle (308.5 (22) v 446.2 (28) N, P = 0.001). PImax at both FRC and RV correlated with maximum oxygen consumption (r = 0.59, P = 0.006, and r = 0.45, P = 0.048 respectively) but not PEmax. There was, however, no significant correlation between PImax and right quadriceps strength. CONCLUSIONS: Respiratory muscle weakness is seen in chronic heart failure. The results suggest that inspiratory muscles are important in determining maximum oxygen consumption and exercise tolerance in these patients. The lack of correlation between respiratory and right quadriceps muscle strength further suggests that the magnitude and time course of respiratory and locomotor muscle weakness may differ in individual patients. Treatment aimed at improving the function of the involved muscle groups may alleviate symptoms. PMID- 7488452 TI - Exercise ventilation after balloon dilatation of the mitral valve. AB - BACKGROUND: Exertional dyspnoea is a limiting symptom in many patients with mitral stenosis but its causes remain incompletely understood. Ventilation during exercise is abnormal in chronic heart failure of all causes and there is increased ventilatory cost of carbon dioxide production. PATIENTS: 23 patients with rheumatic mitral stenosis undergoing percutaneous balloon dilatation of the mitral valve were studied to investigate exercise ventilation. METHODS: Treadmill exercise tests with respiratory gas analysis were performed before and 1 day, 7 days, and 10 weeks after balloon dilatation of the mitral valve. The relation between ventilation (VE) and production (VCO2) was analysed by linear regression. RESULTS: The VE/VCO2 slope was linear in all patients and before balloon dilatation of the mitral valve it correlated inversely with peak minute oxygen consumption (VO2) (rs = -0.47, P < 0.05), exercise duration (rs = -0.66, P < 0.01), and mitral valve area (rs = -0.5, P < 0.05). The VE/VCO2 slope declined acutely after balloon dilatation of the mitral valve (n = 10) (mean (SD) 41 (4) v 36 (2.9), P < 0.05) and did not change again thereafter. At 10 weeks (n = 23) exercise duration (460 (230) v 630 (240) s, P < 0.01) and peak VO2 (12.7 (4.3) v 14.9 (4.8) ml/kg/min, P < 0.05) increased significantly. CONCLUSIONS: Patients with rheumatic mitral stenosis have a similar increase in the VE/VCO2 slope to that of patients with heart failure from other causes. Successful balloon dilatation of the mitral valve is associated with an acute reduction in the exercise VE/VCO2 slope. PMID- 7488453 TI - Heart rate and respiratory rhythm dynamics on ascent to high altitude. AB - OBJECTIVE: To investigate the alterations in autonomic control of heart rate at high altitude and to test the hypothesis that hypoxaemic stress during exposure to high altitude induces non-linear, periodic heart rate oscillations, similar to those seen in heart failure and the sleep apnoea syndrome. SUBJECTS: 11 healthy subjects aged 24-64. MAIN OUTCOME MEASURES: 24 hour ambulatory electrocardiogram records obtained at baseline (1524 m) and at 4700 m. Simultaneous heart rate and respiratory dynamics during 2.5 hours of sleep by fast Fourier transform analysis of beat to beat heart rate and of an electrocardiographically derived respiration signal. RESULTS: All subjects had resting hypoxaemia at high altitude, with an average oxyhaemoglobin saturation of 81% (5%). There was no significant change in mean heart rate, but low frequency (0.01-0.05 Hz) spectral power was increased (P < 0.01) at high altitude. Time series analysis showed a complex range of non linear sinus rhythm dynamics. Striking low frequency (0.04-0.06 Hz) heart rate oscillations were observed during sleep in eight subjects at high altitude. Analysis of the electrocardiographically derived respiration signal indicated that these heart rate oscillations correlated with low frequency respiratory oscillations. CONCLUSIONS: These data suggest (a) that increased low frequency power during high altitude exposure is not simply attributable to increased sympathetic modulation of heart rate, but relates to distinctive cardiopulmonary oscillations at approximately 0.05 Hz and (b) that the emergence of periodic heart rate oscillations at high altitude is consistent with an unstable cardiopulmonary control system that may develop on acute exposure to hypoxaemic stress. PMID- 7488454 TI - Comparison of dual chamber and ventricular rate responsive pacing in patients over 75 with complete heart block. AB - OBJECTIVE: To compare symptoms and exercise tolerance during dual chamber universal (DDD) and ventricular rate response (VVIR) pacing in elderly (> or = 75) patients. DESIGN: Randomised, double blind, crossover study. SETTING: Regional cardiac department. PATIENTS: Twenty elderly patients (mean age 80.5 (1) years) with high grade atrioventricular block and sinus rhythm. Patients with pre existing risk factors for the pacemaker syndrome and chronotropic incompetence were excluded. INTERVENTION: After four weeks of VVI pacing following pacemaker implantation, patients underwent consecutive two week periods of VVIR and DDD pacing. MAIN OUTCOME MEASURES: Patient preference, symptom scores, "daily activity exercises," and perceived level of exercise (Borg score). RESULTS: Eleven patients preferred DDD mode to either VVI or VVIR mode. Mean (SE) total symptom scores during VVI, VVIR, and DDD pacing were 5.9 (1.1), 6.1 (1.0), and 3.5 (0.9) respectively (P < 0.01). The corresponding mean (SE) pacemaker syndrome symptom scores were 4.8 (0.7), 5.2 (0.8), and 2.9 (0.8) (P < 0.05). Symptom scores during VVI and VVIR pacing were not significantly different. Exercise performance and Borg scores were significantly worse during VVI pacing compared with VVIR or DDD pacing but did not significantly differ between VVIR and DDD modes. CONCLUSIONS: In active elderly patients with complete heart block both DDD and VVIR pacing are associated with improved exercise performance compared with fixed rate VVI pacing. The convenience and reduced cost of VVIR systems, however, may be offset by a higher incidence of the pacemaker syndrome. In elderly patients with complete heart block VVIR pacing results in suboptimal symptomatic benefit and should not be used instead of DDD pacing. PMID- 7488455 TI - Atrial electromechanical sequence in normal subjects and patients with DDD pacemakers. AB - OBJECTIVE: To assess the effect of right atrial appendage pacing on atrial electromechanical interrelations in patients with DDD pacemakers. DESIGN: Prospective study by M mode echocardiogram, Doppler echocardiogram, and apexcardiogram, along with electrocardiogram and phonocardiogram. SETTING: Tertiary cardiac referral centre. PATIENTS: 20 patients with DDD pacemakers and 20 age matched normal controls. RESULTS: Age, RR interval, atrial size, left ventricular size, and fractional shortening were similar in the two groups. Atrial electromechanical delay (the time from the onset of P wave or atrial pacing spike on ECG to the onset of atrial contraction on M mode echogram) was 68 (SD 7) ms at the lateral site of right atrium, 82 (9) ms at the central fibrous body, 93 (11) ms at the lateral site of left atrium in normals. In patients with DDD pacing, however, this delay increased to 85 (22) ms, 117 (23) ms, and 138 (25) ms respectively (all P < 0.01). Interatrial mechanical delay (the time from the onset of right atrial motion to the onset of the left) increased from 25 (6) ms in normal controls to 53 (18) ms in patients (P < 0.01). Intra-atrial mechanical dispersion (the time from the earliest to the latest onset of regional atrial motion around the atrioventricular ring) in the right atrium increased from 6 (2) ms in normals to 19 (2) ms in patients (P < 0.01), but it remained unchanged in the left atrium (6 (2) ms in normal controls v 7 (2) ms in patients, P > 0.05). Peak atrial shortening rate was not different between the two groups. Differences of atrial electromechanical activity between the two groups were also reflected on Doppler echocardiogram and apexcardiogram. CONCLUSIONS: Right atrial appendage pacing disturbs the normal coordinate sequence of right atrial mechanical activity and leads to a striking and variable increase in intra-atrial conduction time as well as in interatrial conduction time. Left atrial contraction remains synchronous although the timing of the start of its contraction was delayed. These values can be determined in individual patients to allow optimal setting of DDD pacemakers. PMID- 7488457 TI - Optimal analysis of the signal averaged P wave in patients with paroxysmal atrial fibrillation. AB - OBJECTIVE: To define the ability of analysis of the signal averaged P wave to identify patients with paroxysmal atrial fibrillation (AF) and establish whether differences in quantitative variables between patients and controls are due to concurrent cardiopulmonary disease, greater atrial dimension, or to unrelated changes in atrial electrophysiology. DESIGN: An observational parallel group study. SETTING: Cardiac department of a busy district general hospital. PATIENTS: 58 participants without cardiopulmonary disease (24 with paroxysmal AF and 34 controls, group A) and 57 with cardiac or respiratory conditions (31 with paroxysmal AF and 26 controls, group B). Mean (range) age of patients was 54 (25 71) and controls 53 (34-78) for group A and 65 (45-81) and 62 (36-78) respectively for group B. Left atrial size was similar in patients and controls in each group (mean (SEM)) group A: 2.39 (0.1) v 2.19 (0.07) cm; group B: 2.51 (0.10) v 2.71 (0.12) cm). MAIN OUTCOME MEASURES: Analysis of the P wave after P wave-specific signal averaging. Filtered P wave duration and spatial velocity were calculated. Energies contained in frequency bands from 20, 30, 40, 60, and 80 to 150 Hz after spectral analysis were expressed as absolute values (P20, P30 etc) and ratios of high to low frequency energy (PR20, PR30, etc). RESULTS: Duration and peak spatial velocity were increased in patients with paroxysmal AF (median (interquartile range) duration group A: 144 (137-155) v 136 (129-143) ms, P = 0.007; group B: 155 (144-159) v 142 (136-151) ms, P = 0.002; peak spatial velocity group A: 16.5 (14.1-21.2) v 14.5 (11.7-18.1) mV/s, P = 0.02; group B: 18.9 (14.8-21.8) v 14.3 (12.6-17.6) mV/s, P = 0.01). Energy contained in frequency bands from 20, 30, 40, 60 and 80 to 150 Hz was expressed as absolute values (P20, P30, P40, P60, and P80) and percentage energy ratios. P30, P60, and P80 were significantly greater in patients with AF in group A (for example P60: 3.9 (3.0-5.3) v 3.1 (2.0-4.3) microV2.s, P = 0.02) and P20, P30, and P40 were increased in those with AF in group B (for example P40: 16.7 (9.9-20.8) v 10.8 (8.1-14.8) microV2.s, P = 0.02). A score developed from logistic regression analysis of duration and P60 identified patients with paroxysmal AF with a sensitivity of 81% and specificity of 73%. CONCLUSIONS: Increased P wave duration and magnitude are associated with paroxysmal AF with and without additional cardiopulmonary disease. The discriminant ability of the signal averaged P wave is improved by analysis of duration and a magnitude variable. These results invite prospective assessment of the ability of the signal averaged P wave to predict paroxysmal AF in unselected patients. PMID- 7488458 TI - Early diastolic left ventricular inflow pressures in normal subjects and patients with dilated cardiomyopathy. Reconstruction from pulsed Doppler echocardiography. AB - OBJECTIVE: To estimate early diastolic left ventricular inflow pressures in normal subjects and patients with dilated cardiomyopathy, and thus to assess the potential effect of restoring forces. METHODS: Early diastolic left ventricular inflow pressures were reconstructed using the ventricular blood as an accelerometer, by measuring velocity at 1 cm intervals within the left ventricle from mitral ring to apex by pulsed Doppler echocardiography, and differentiating the records to obtain the acceleration. Aortic component of second heart sound (A2) was used to fix relative timings. The local pressure gradient was determined from the acceleration at each level, and the total pressure drop during the acceleration (+ peak PD) and deceleration (- peak PD) phases of the filling interval were determined by summing the local increments. The total stroke volume (SV) at the left ventricular outflow tract and the mitral stroke distances (MSD) were also determined, using the time-velocity integral at mitral ring level. Effective flow orifice area was thus SV/MSD. Inflow jet width across the mitral valve was estimated by cross sectional colour Doppler flow mapping. PATIENTS: 32 patients with dilated cardiomyopathy with a dominant mitral E or summation wave, and 24 normal subjects of similar ages. RESULTS: Normal + peak PD was 3.9 (SD 0.7) v 7.4 (2.2) mm Hg in dilated cardiomyopathy (P < 0.01). Normal - peak PD was 2.5 (0.9) v 5.6 (2.8) mm Hg in cardiomyopathy (P < 0.01). Normal effective flow orifice area was 5.9 (1.3) v 1.9 (0.8) [range 0.9 approximately 3.7] cm2 in cardiomyopathy (P < 0.01). This corresponded to 71 (18)% of the end systolic cavity cross section in normals v 11 (6)% in dilated cardiomyopathy (P < 0.01). Normal cross sectional colour inflow jet width was 2.7 (0.3) v 1.5 (0.4) cm in cardiomyopathy (P < 0.01). The jet width correlated with flow width calculated from effective flow orifice area (r = 0.82, P < 0.01). CONCLUSIONS: (1) Total early diastolic positive and negative peak pressure drop are normally low, so that significant negative left ventricular pressures are not needed to explain normal resting early diastolic mitral flow velocities. (2) These low pressure drops are only possible with a large effective orifice area approaching end systolic left ventricular cavity area. (3) Atrioventricular pressure drops are much greater in dilated cardiomyopathy, where increased inflow accelerations are due to reduced effective flow orifice area. These disturbances will impair filling independently of any abnormality of relaxation or compliance. PMID- 7488456 TI - Favourable outcome in idiopathic ventricular fibrillation with treatment aimed at prevention of high sympathetic tone and suppression of inducible arrhythmias. AB - OBJECTIVE: In the absence of an obvious cause for cardiac arrest, patients with idiopathic ventricular fibrillation are difficult to manage. A subset of patients has inducible arrhythmias. In others sympathetic excitation plays a role in the onset of the cardiac arrest. This study evaluates a prospective stepped care approach in the management of idiopathic ventricular fibrillation, with therapy first directed at induced arrhythmias and secondly at adrenergic trigger events. SETTING: University Hospital. PATIENTS: 10 consecutive patients successfully resuscitated from idiopathic ventricular fibrillation. INTERVENTIONS: Programmed electrical stimulation to determine inducibility, followed by serial drug treatment. Assessment of pre-arrest physical activity and mental stress status by interview, followed by beta blockade. Cardioverter-defibrillator implantation in non-inducible patients not showing significant arrest related sympathetic excitation. MAIN OUTCOME MEASURE: Recurrent cardiac arrest or ventricular tachycardia. RESULTS: Five patients were managed with serial drug treatment and four with beta blockade. In one patient a defibrillator was implanted. During a median follow up of 2.8 years (range 6 to 112 months) no patient died or experienced defibrillator shocks. One patient had a recurrence of a well tolerated ventricular tachycardia on disopyramide. CONCLUSIONS: Idiopathic ventricular fibrillation may be related to enhanced sympathetic activation. Prognosis may be favourable irrespective of the method of treatment. Whether the present approach enhances prognosis of idiopathic ventricular fibrillation remains to be determined. However, it may help to avoid potentially hazardous antiarrhythmic drugs or obviate the need for implantation of cardioverter defibrillators. PMID- 7488459 TI - Myocardial calcium-independent nitric oxide synthase activity is present in dilated cardiomyopathy, myocarditis, and postpartum cardiomyopathy but not in ischaemic or valvar heart disease. AB - OBJECTIVE: To determine the activity of the calcium-dependent constitutive (cNOS) and calcium-independent inducible nitric oxide (iNOS) synthases in heart tissue from patients with different cardiac diseases. PATIENTS AND DESIGN: Endomyocardial biopsy specimens were obtained from patients with dilated hearts (by echocardiography and ventriculography) and normal coronary arteries (by selective angiography). Recognised clinical, radiological, and histopathological criteria were used to diagnose non-inflammatory dilated cardiomyopathy (DCM) (n = 6), inflammatory cardiomyopathy (ICM) (n = 5), and peripartum cardiomyopathy (PPCM) (n = 3). Comparative groups were chosen with similarly dilated hearts caused by ischaemic (n = 5) or valvar disease (n = 4), and, in addition, non dilated hearts with ischaemic (n = 5) and valvar (n = 3) disease. Venous blood was taken at the time of myocardial biopsy for assay of plasma tumour necrosis factor alpha (TNF alpha). RESULTS: Myocardial tissue from patients with DCM, ICM, and PPCM showed considerable iNOS activity (16.8 (2.7) pmol citrulline/mg protein/min) with little or no cNOS activity (1.3 (0.9) pmol citrulline/mg protein/min). In contrast, myocardial tissue from patients with both dilated and non-dilated hearts of ischaemic or valvar aetiology showed cNOS and little, if any, iNOS activity (dilated--cNOS 11.7 (2.4) and iNOS 0.8 (0.6) pmol citrulline/mg protein/min; non-dilated--cNOS 12.1 (1.8) and iNOS 1.4 (0.8) pmol citrulline/mg protein/min). Plasma TNF alpha was detectable only in patients with inflammatory DCM. CONCLUSIONS: These results support the hypothesis the generation of nitric oxide by iNOS accounts for some of the dilatation and impaired contractility associated with inflammatory and non-inflammatory dilated cardiomyopathy and peripartum cardiomyopathy. PMID- 7488460 TI - Reductions in muscle sympathetic nerve activity after long-term metoprolol for dilated cardiomyopathy: preliminary observations. AB - OBJECTIVE: To determine whether efferent muscle sympathetic nerve activity diminishes in subjects with dilated cardiomyopathy who improve after long term treatment with metoprolol. METHODS: Microneurographic, echocardiographic, plethysmographic, and neurohumoral data were obtained immediately before and 20 months after the addition of beta blockade in seven subjects with idiopathic dilated cardiomyopathy with clinical deterioration despite conventional treatment. RESULTS: Six subjects (three men, three women, aged 24-62 years) were restudied after a mean (SEM) of 20 (2.4) months treatment with metoprolol (45.8 (2.6) mg/d). Long term treatment was associated with decreases in left ventricular end diastolic and end systolic diameter (P < 0.005), left ventricular mass index (P < 0.05), and atrial natriuretic factor (P < 0.05), and increases in fractional shortening (P < 0.05) and mean blood pressure (P < 0.05). There was a 50% reduction in peroneal muscle sympathetic nerve activity (from 49.2 (10.1) to 24.5 (4.7) bursts/min; (P < 0.005) and a 62% decrease in calf vascular resistance (from 56.2 (4.4) to 21.2 (5.7) units; P < 0.005). This reduction in pulse synchronous nerve activity was not simply a function of bradycardia (heart rate fell from 94.2 (4.6) to 62.8 (5.7) beats/min; P < 0.005) since muscle sympathetic burst incidence also decreased (from 51 (8.7) to 37.5 (5.2) bursts/100 heart beats; P < 0.05). Similar haemodynamic improvement was observed in the seventh subject, who was switched to sotalol 200 mg/d and restudied after 20 months, but burst frequency was 50% higher and calf vascular resistance 93% higher. CONCLUSIONS: Muscle sympathetic nerve activity and calf vascular resistance decrease in patients with dilated cardiomyopathy who improve after long term treatment with metoprolol. Inhibition of central sympathetic outflow may be one mechanism by which metoprolol benefits such subjects. PMID- 7488461 TI - Morphology of the sinus node in human and mouse hearts with isomerism of the atrial appendages. AB - BACKGROUND: The location of the sinus node is known to be at best abnormal, or at worst unknown, in patients with isomerism of the morphologically left atrial appendage. In contrast, the sinus node is known to be an excellent histological marker of the morphologically right appendage, being duplicated in those with right isomerism. The aim of the study was to investigate this condition further in fetal human and mouse hearts. METHODS: Serial histological sections of the area anticipated to contain the sinus node were studied in hearts with isomerism of the atrial appendages taken from 14 human fetuses and 13 iv/iv mice, using 12 mouse hearts with normally arranged or mirror imaged atrial chambers for controls. RESULTS: All hearts with isomerism of the right appendages (two human and four mouse) had bilateral sinus nodes. The cases with isomerism of the left appendages (12 human and nine mouse) showed absence of a recognisable sinus node except in four cases (19%) in which a small remnant of the node was found. In three of these cases, it was related postero-inferiorly to the superior cavoatrial junction. CONCLUSIONS: The concept of isomerism of the atrial appendages is endorsed by findings on the morphology of the sinus node, this being the most reliable histological criterion for existence of a morphologically right atrium. A small proportion of hearts with left isomerism had a structure resembling the sinus node, but it was hypoplastic and displaced postero inferiorly, distant from its expected position had the hearts possessed an incompletely formed morphologically right appendage. PMID- 7488462 TI - Echocardiographic-anatomical correlations in aorto-left ventricular tunnel. AB - OBJECTIVE: To investigate the echocardiographic, morphological, and histological appearances of aorto-left ventricular tunnel observed in four fetal hearts and compare the findings with those reported in older patients with the malformation. BACKGROUND: Previous studies have concentrated on clinical features of the malformation from birth to adult life and have speculated on either its embryological formation or its acquisition during late intrauterine life. The presentation of a large series of cases in fetal life is a unique opportunity to study the malformation at an early stage in its natural course. METHODS: A retrospective study was performed of four cases of aorto-left ventricular tunnel discovered among 872 cases of congenital abnormalities diagnosed at a tertiary centre for fetal echocardiography. Detailed echocardiographic and anatomical observations were made of the malformation as identified during fetal life. The precise anatomical arrangement was determined and compared with previous descriptions found in journals published in English. RESULTS: In fetal life, as after birth, the malformation is characterised by enlargement and hypertrophy of the left ventricle, enlargement of the aortic root, and free regurgitation at the level of the aortic valve. Anatomical abnormalities are found at the aortic ventriculoarterial and sinutubular junctions as well as in the intervening aortic wall. These are unrelated to necrosis, ischaemia, or the presence of mucopolysaccharides. CONCLUSIONS: The lesion is a developmental abnormality that should be reliably diagnosed by fetal echocardiography combined with colour flow Doppler echocardiography during the mid-trimester. The exact anatomical relations clarified by this study are pertinent to diagnosis and subsequent surgical correction. PMID- 7488463 TI - Mortality in participants and non-participants of a multifactorial prevention study of cardiovascular diseases: a 28 year follow up of the Helsinki Businessmen Study. AB - OBJECTIVE: To investigate pretrial risk factors and long term mortality (1964 1992) in participants and non-participants of a multifactorial primary prevention trial. DESIGN: A prospective study among 3313 initially healthy businessmen. During the 1960s (1964 onwards), 3490 healthy male business executives born between 1919 and 1934 participated in voluntary health checks at the Institute of Occupational Health in Helsinki. From that period cardiovascular disease (CVD) risk factors were available in 3313 men. In the beginning of the 1970s these men were invited to join a multifactorial primary prevention trial of CVD. Six groups were formed: (I) healthy participants in a high risk intervention group (n = 612), and (II) their randomised control group (n = 610); (III) a non-participant low risk group (n = 593); (IV) an excluded group with signs of CVD (n = 563); (V) a refused group (n = 867); and (VI) dead (n = 68). Groups I and II participated in the five year prevention trial which started in 1974. Other groups were followed up through registers, with no personal contact. MEASUREMENTS: Cardiovascular risk factors during the 1960s. Mortality follow up using national registers up to 31 December, 1992. MAIN RESULTS: Baseline risk factors were lowest in the low risk group, highest in the excluded group, intermediate and comparable in other groups. Eighteen-year (1974-1992) mortality (per 1000) was 79.3, 106.6, 155.2, 179.9, and 259.3 in the low risk, control, intervention, refused, and excluded groups, respectively (P < 0.001). In the whole population of 3313 men, the 28-year (1964-1992) total (n = 577) and coronary deaths (n = 199) were significantly predicted by smoking, blood pressure, and cholesterol; cancer deaths (n = 163) by smoking only; and violent deaths (n = 83) by none of the risk factors. One-hour postload glucose was significantly associated with total mortality in the intervention group only. When the intervention and control groups were included in the same model, the effect of group on total mortality tended to be dependent on the 1 h blood glucose value (P = 0.06 for the group by 1 h glucose interaction term). CONCLUSION: The traditional risk factors (smoking, blood pressure, and cholesterol) are significantly associated with 28-year mortality in this high social class population with previous health education. Conversely, a "clustering" of low risk factors predicted low total, coronary, and cancer mortality. The findings on 1 h blood glucose suggest that factors related to glucose tolerance explain in part the excess mortality in the intervention group compared with the control group. PMID- 7488465 TI - In vitro validation of right ventricular volume measurement by three dimensional echocardiography. AB - OBJECTIVE: Evaluation of ability of three dimensional echocardiography to accurately assess right ventricular volumes in vitro. METHODS: Silicone casts of normal human right ventricles were examined. Each was filled with three different volumes of water to yield 15 different measurements. The casts were examined in a waterbath with three dimensional echocardiography using a 7.5 MHz ultrasound probe mounted in a scan frame. It was steered by a stepper motor, which moved the probe in steps of 0.25 mm over a distance of 5.9 cm inside the frame, acquiring an image at each step. 236 parallel slices of the cast were thus obtained, forming the three dimensional dataset. The longest axis of the right ventricular volume was defined and the area of perpendicular 1 mm thick slices was outlined manually to calculate the area of each slice. This was multiplied by the slice thickness to obtain the volume of each slice; the respective volumes were added to obtain the volume of the whole cast. RESULTS: The casts had a median volume of 31.1 (23) ml (range 15-100); three dimensional echocardiography gave a median volume of 29.0 (21.7) ml (15.7-91.7). Interobserver variability was 4.5% (0.4% 13.6%) and intraobserver variability 4.3% (0.2%-9.3%). Correlation between real cast volumes and volumes measured by three dimensional echocardiography was 0.99 (y = 1.08 x -0.16) with an SEE of 2.7 ml. Limits for agreement between methods ranged from -3.1 ml to 8.3 ml. In 14 of the 15 measurements, volume by three dimensional echocardiography was smaller than real volume, with the mean difference being 7.4% (2.8%-19.5%). This may be due to the thickening of surfaces of structures when imaged by ultrasonography. CONCLUSION: Right ventricular volumes can accurately be determined by three dimensional echocardiography. PMID- 7488464 TI - Combined mechanical and thrombolytic treatment for totally occluded bypass grafts. AB - Totally occluded venous grafts are usually less amenable to mechanical reperfusion alone (for example percutaneous transluminal coronary angioplasty, PTCA) because of the large mass of thrombotic material within the graft. A combined approach using mechanical and thrombolytic treatment might therefore be more successful. Twenty one patients (20 males, one female) with a mean age of 64.5 (SD 5.6) years underwent angiography because of crescendo or unstable angina (n = 19) or myocardial infarction (n = 2) at a mean of 21.7 (18.6) days after onset of symptoms (range 1-60). All patients had had coronary artery bypass grafting (CABG) at a mean of 8.02 (4.02) years (range 0.3-13 years) before the current admission. At catheterisation, totally occluded venous bypass grafts to the left anterior descending coronary artery or diagonal (n = 10), marginal (n = 6), or right coronary artery (n = 5) were found. A combination of PTCA and thrombolytic treatment (in eight patients extended thrombolysis for 24 hours) was successful in reopening the venous graft in 16/21 patients (76.2%). Immediate complications included femoral haematoma (4), distal embolisation (3), and infection in one patient. Out of 13 patients catheterised within three months, two had reoccluded, seven had restenosis, while four had patent grafts. Recurrent PTCA (at least once more) was done in eight patients. At long term follow up of a mean of 26.7 (21.6) months (range 4-75 months), four patients were asymptomatic, eight still suffered from mild stable angina, while three had recurrent hospital admissions and needed a second coronary artery bypass. A combination of thrombolytic treatment and PTCA is a feasible and practised approach to recanalise recently occluded venous bypass grafts. PMID- 7488466 TI - Chest pain during exercise as first manifestation of Friedreich's ataxia. AB - Cardiac function is affected in up to 90% of patients with Friedreich's ataxia, the most common spinocerebellar degenerative disease. Friedreich's ataxia typically causes motor abnormalities of the extremities, mainly impairing walking and the coordination of the legs and arms. The myocardium is affected at a later stage of the disease. The extent and timing of myocardial involvement determines the clinical course. Some patients have no cardiac symptoms and cardiac involvement can be established only by electrocardiographic or echocardiographic examination. In addition some pathological studies have found evidence of coronary abnormalities, mainly in the small vessels. There are no reports that such lesions cause angina. In a 16 year old patient chest pain on exercise had been the presenting symptom of Friedreich's ataxia at the age of 9. Considerable alterations in ventricular repolarisation on the electrocardiogram suggested a congenital coronary abnormality or hypertrophic myocardiopathy. The results of a Doppler echocardiography, Holter monitoring, and a haemodynamic study with coronary arteriography were all normal. An exercise test when the boy was 13 indicated significant changes in ventricular repolarisation. Myocardial scintigraphy (99mTc-MIBI) at that time, however, was normal. He improved slightly when he was treated with verapamil. When he was 15 neurological symptoms developed and Friedreich's ataxia was diagnosed. Typical angina during exercise seems to have been the first symptom of Friedreich's ataxia. PMID- 7488467 TI - Excessive numbers of skin cancers and pre-malignant skin lesions in an Australian heart transplant recipient. AB - One and a half years after heart transplantation an Australian man developed his first skin cancer. In the period until his death, 7 years after the transplantation, 34 histologically confirmed squamous cell carcinomas developed, 17 intra-epidermal carcinomas, and 9 basal cell carcinomas. Most skin cancers were confined to chronically sun exposed sites. Exposure to sunlight and human papillomavirus are important factors in the development of skin cancer in renal transplant recipients, and further studies are needed to establish the role of these risk factors in cardiac transplant recipients. PMID- 7488468 TI - Direct evidence of patent foramen ovale as a route for paradoxical embolism. PMID- 7488469 TI - Probable right ventricular dysplasia and patent foramen ovale presenting with cyanosis and clubbing in a patient with characteristics of Noonan syndrome. AB - Probable right ventricular dysplasia and a patent foramen ovale resulted in cyanotic heart disease in a patient with some characteristics of Noonan syndrome. PMID- 7488470 TI - How do the clinical findings in patients with pericardial effusions influence the success of aspiration? PMID- 7488471 TI - Is thrombolytic therapy really better than conventional treatment in acute inferior myocardial infarction? PMID- 7488472 TI - Is thrombolytic therapy really better than conventional treatment in acute inferior myocardial infarction? PMID- 7488473 TI - Research in anaesthesia--the key to the future. PMID- 7488474 TI - Hormonal effects of thoracic extradural analgesia for cardiac surgery. AB - We have investigated the hormonal and metabolic effects of thoracic extradural analgesia with bupivacaine in addition to sufentanil 20 micrograms kg-1 in nine patients undergoing coronary artery bypass surgery. A control group received general anaesthesia alone. The catecholamine response was inhibited for 24 h after surgery in patients who had received extradural analgesia, and the cortisol response was suppressed during cardiopulmonary bypass. Blood glucose values were unchanged until 24 h after surgery in the extradural group. There were no significant differences in cardiac index between the two groups of patients, although heart rate and mean arterial pressure decreased before surgery in patients who received extradural analgesia. The benefits of this technique in terms of a reduction in postoperative morbidity remain to be determined. PMID- 7488475 TI - Interaction of extradural morphine and lignocaine on ventilatory response. AB - We have evaluated the effects of lumbar extradural morphine and lignocaine on the ventilatory response to carbon dioxide. Twenty-four female patients were allocated randomly to receive extradural morphine 2 mg (group M), 2% lignocaine 10 ml (group L) or a combination of morphine 2 mg and 2% lignocaine 10 ml (group ML). On the day before surgery, resting ventilatory values including minute volume (VE) and tidal volume (VT), and ventilatory response to progressive hyperoxic hypercapnia (VE/PE'CO2) were measured. On the day of surgery, the same measurements were repeated 30 min after extradural injection. Ventilatory values at rest were not altered after extradural injection. Mean VE/PE'CO2 decreased significantly after extradural morphine (P = 0.002) and increased (P = 0.011) after extradural lignocaine. Mean VE 7.3 (VE at PE'CO2 7.3 kPa) decreased significantly after extradural morphine (P < 0.001) and increased after extradural lignocaine (P = 0.047). Extradural morphine and lignocaine did not significantly alter mean VE/PE'CO2 and mean VE 7.3: 14.6 (95% confidence intervals 12.1-17.1) to 15.3 (13.1-17.6) litre min-1 kPa-1 and 22.8 (18.1-27.5) to 22.8 (17.3-28.3) litre min-1, respectively. We conclude that extradural co administration of morphine and lignocaine did not increase the risk of respiratory depression associated with morphine. PMID- 7488476 TI - A progressive care programme for prolonged ventilatory failure: analysis of outcome. AB - Forty consecutive patients who could not be weaned from mechanical ventilation in the intensive care unit (ICU) entered a multidisciplinary progressive care programme (PCP). The mean number of hours per day of ventilatory support was 19.9 at the time of transfer but only 6.7 at discharge. Eleven patients did not require ventilation after discharge, 24 received ventilation non-invasively and only three via a tracheostomy. Survival at discharge from hospital was 90% compared with the predicted survival of 53% from the Apache II scores on admission to the ICU. Seventy-six percent were alive 1 yr after discharge and 80% of patients were discharged directly from the PCP to their homes. Mental and emotional scores in a quality of life questionnaire (SF 36) were normal, but physical function remained limited. PMID- 7488477 TI - A prospective, randomized comparison of interpleural and paravertebral analgesia in thoracic surgery. AB - We have undertaken a prospective, randomized comparison of the superficially similar techniques of interpleural and paravertebral (extrapleural) analgesia in 53 patients undergoing posterolateral thoracotomy. Local anaesthetic placed anterior to the superior costotransverse ligament and posterior to the parietal pleura produces a paravertebral block and instilled between the parietal and visceral pleurae produces an interpleural block. Patients received preoperative and postoperative continuous bupivacaine paravertebral blocks in group 1 and interpleural blocks in group 2. Premedication comprised diclofenac and morphine, and after operation all patients had regular diclofenac and patient-controlled morphine (PCM). Analgesia was assessed by visual analogue pain scores (VAS), PCM requirements, ratio of preoperative to postoperative spirometric values (PFT), rates of postoperative respiratory morbidity (PORM) and hospital stay, all recorded by blinded observers. Eight patients were withdrawn and data from 45 patients were analysed. Patient characteristics, surgery, VAS scores and PCM use were similar in both groups. PFT were significantly better (P = 0.03-0.0001) in group 1, and PORM was lower and hospital stay approximately 1 day less in this group. Five patients in group 2 became temporarily confused, probably because of bupivacaine toxicity (P = 0.02). We conclude that bupivacaine deposited paravertebrally produced greater preservation of lung function and fewer side effects than bupivacaine administered interpleurally. PMID- 7488478 TI - Comparison of i.m. and local infiltration of ketorolac with and without local anaesthetic. AB - We have studied postoperative analgesia in 32 patients undergoing outpatient repair of inguinal hernia. All patients received a standardized general anaesthetic of thiopentone followed by halothane or isoflurane in 70% nitrous oxide and oxygen delivered by face mask or laryngeal mask with spontaneous ventilation. No patient received premedication, opioids or neuromuscular blockers. Before wound closure the surgeon infiltrated 20 ml of a study solution into the wound. The solution contained ketorolac 30 mg in saline, 0.25% bupivacaine and ketorolac 30 mg, 0.25% bupivacaine or saline (control group). The control group received ketorolac 60 mg i.m. at the same time. Pain scores were measured 2, 6 and 24 h after operation. Pain scores for all three active groups were significantly less (P < 0.05) than the control group at all times. There were no significant differences in pain scores at any time between the three study groups. Wound infiltration with ketorolac 30 mg in saline, 0.25% bupivacaine or ketorolac 30 mg with 0.25% bupivacaine provided equivalent analgesia. Wound infiltration with ketorolac 30 mg in saline provided analgesia superior to that of ketorolac 60 mg i.m. PMID- 7488480 TI - Effects of inspired gas composition during anaesthesia for abdominal hysterectomy on postoperative lung volumes. AB - We have studied 51 patients who were allocated randomly and prospectively to receive either 100% oxygen (n = 16), 70% nitrous oxide in oxygen (n = 18) or 30% oxygen in nitrogen (n = 17) as the inspired gas during anaesthesia for abdominal hysterectomy. Lung volumes were measured before and after surgery. TLC, VC, FVC and FEV1 but not RV or FRC were reduced after surgery. There were no significant differences between the three treatment groups in any of the lung volumes measured. We conclude that absorption atelectasis during anaesthesia is not the main cause of perioperative changes in lung volume after abdominal hysterectomy. Any effect of the inspired gas is likely to be of limited clinical significance. PMID- 7488481 TI - Influence of inspired nitrogen concentration during anaesthesia for coronary artery bypass grafting on postoperative atelectasis. AB - Pulmonary collapse is a common problem after coronary artery bypass graft surgery (CABG). If absorption atelectasis during anaesthesia is an important mechanism in the genesis of pulmonary collapse after CABG, the addition of nitrogen to the inspired gas during anaesthesia should reduce the amount of postoperative collapse. We studied 30 patients who were allocated randomly and prospectively to receive either 100% oxygen or an oxygen-air mixture as the inspired gas during anaesthesia for CABG. Lung volumes, PaO2, and an x-ray atelectasis score were measured before and after surgery to assess the degree of atelectasis. There were no significant differences between the two treatment groups in any of these measurements. PMID- 7488479 TI - Effect of inspired oxygen on the cardiovascular effects of protamine after cardiopulmonary bypass. AB - We have performed a randomized prospective study of the effects of inspired oxygen fraction (FIO2) on the haemodynamic changes after protamine infusion. Thirty-four patients undergoing first time coronary artery bypass surgery were allocated randomly to receive either an FIO2 of 1.0 (group O) or 0.35 (group A) after cardiopulmonary bypass. Before and after infusion of protamine, haemodynamic measurements were obtained, including mean arterial pressure (MAP), mean pulmonary artery pressure, central venous pressure, pulmonary capillary wedge pressure (PCWP), cardiac index (CI), pulmonary vascular resistance index (PVRI) and systemic vascular resistance index (SVRI). In group O, there were increases in mean MAP (8%), PVRI (48%) and SVRI (18%), and decreases in mean CI (10%) and PCWP (15%). Group A showed changes of 0%, -8%, -6%, +3% and +32%, respectively. We found a significant difference between groups in changes in PVRI (P < 0.0001), SVRI (P < 0.01), CI (P < 0.05) and PCWP (P < 0.001). During infusion of protamine, 31% of patients in group O and 6% of patients in group A had a decrease in systolic arterial pressure to less than 80 mm Hg (ns, chi square test). These observations suggest that FIO2 alters the haemodynamic effects of protamine. PMID- 7488482 TI - The auditory evoked potential and paediatric anaesthesia. AB - We have examined the raw EEG activity and auditory evoked responses (AER) in 35 children, aged 3 days to 13 yr (median 1.5 yr), undergoing general anaesthesia for routine surgery. Binaural stimuli were presented at a frequency of 6.12 Hz and the EEG was recorded and stored using the Northwick Park auditory evoked response software. AER were generated by averaging 512 sweeps each of 125-ms duration. In children less than 2 yr of age, the AER was often irregular, whereas children older than 2 yr had AER patterns similar to those in adults. In children less than 2 yr, regular artefact activity was superimposed on the background EEG which was at the same frequency as the instantaneous heart rate and which was often identifiable as the ECG. We conclude that the AER may be unreliable in children less than 2 yr of age, and modification of current methodology may be required if this technique is to become useful in paediatric anaesthetic practice. PMID- 7488483 TI - Pharmacokinetics of 1R-cis 1'R-cis atracurium besylate (51W89) and plasma laudanosine concentrations in health and chronic renal failure. AB - To ascertain the effects of chronic renal failure on the pharmacokinetics of 1R cis 1'R-cis atracurium besylate (a stereoisomer, designated 51W89), we gave a bolus dose of 0.1 mg kg-1 (2 x ED95) to 17 patients with end-stage renal failure and to 15 patients with normal renal function undergoing elective surgery. All patients received thiopentone, fentanyl and midazolam i.v. and 70% nitrous oxide in oxygen. Blood samples were obtained over 8 h and plasma analysed for 51W89 and laudanosine concentration, using high pressure liquid chromatography. A two compartment model was fitted to the 51W89 plasma concentration data using the NONMEM program, to estimate pharmacokinetic variables and to determine the influence of renal failure, age, weight and sex. Clearance of 51W89 was found to be reduced by 13% in renal failure. The typical value of T1/2 beta was 4.2 min longer in renal failure than in the healthy patients (34.2 vs 30.0 min, P < 0.005). In the healthy patients, clearance of 51W89 was greater in males, but it decreased with increasing age by approximately 1.5 ml min-1 yr-1. Mean plasma laudanosine concentrations were significantly higher in the renal failure group; nevertheless, they were approximately one-tenth of those reported after atracurium. PMID- 7488484 TI - Onset of neuromuscular block after tourniquet inflation: comparison of suxamethonium with vecuronium. AB - To determine the influence of circulatory factors on onset of neuromuscular block, we have measured twitch height in an arm with a tourniquet inflated during onset and compared this with data from a control arm in 20 patients under fentanyl-thiopentone-nitrous oxide-isoflurane anaesthesia. Patients were allocated randomly to receive either vecuronium 0.1 mg kg-1 (n = 10) or suxamethonium 1 mg kg-1 (n = 10). The EMG response of the first dorsal interosseous to single twitch stimulation of the ulnar nerve every 10 s was recorded in both arms. When neuromuscular block was 20% (i.e. twitch height was 80% of control), the tourniquet was inflated to a pressure of 250 mm Hg. It was deflated 5 min later. In the vecuronium group, the rate of onset did not differ in both arms and mean maximum block was 95 (SD 4)% in the tourniquet arm, which was not different from 99 (2)% in the perfused arm. In the suxamethonium group, the presence of a tourniquet decreased the rate of onset by 66%. Maximum block was only 74 (20)% in the tourniquet arm compared with 97 (5)% in the perfused arm (P < 0.05). The difference in maximum neuromuscular block between arms was 4 (3)% in the vecuronium group and 22 (17)% in the suxamethonium group (P < 0.01). We conclude that during onset, neuromuscular block continued to increase in spite of interruption of blood flow, and this increase was greater with vecuronium than with suxamethonium.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7488485 TI - EEG arousal during laryngoscopy and intubation: comparison of thiopentone or propofol supplemented with nitrous oxide. AB - We studied EEG arousal after laryngoscopy and intubation with standardized bolus induction of anaesthesia. Twenty patients were prospectively allocated randomly to induction with propofol 3 mg kg-1 (n = 10) or thiopentone (6 mg kg-1 (n = 10) and 50% nitrous oxide in oxygen. Neuromuscular block was produced with vecuronium 0.2 mg kg-1 given 30 s after induction. Three minutes after induction, laryngoscopy was performed for 60 s, with intubation at 3 min 30 s, and study end at 5 min. Nociception to laryngoscopy and intubation was followed by loss of low (relative delta activity change: thiopentone -30%, propofol -7%; P < 0.05) and a shift to higher frequency EEG activity (beta activity change: thiopentone +647%, propofol +61%; P < 0.05). This EEG arousal was greater in the thiopentone group, despite the fact that EEG depression was similar to that produced by propofol before laryngoscopy and intubation. Propofol and thiopentone in combination with nitrous oxide had similar cortical depressant effects, but propofol appeared to depress subcortical nociceptive processing more than thiopentone. While the degree of cortical EEG depression seems less useful for predicting reaction to subsequent nociception, EEG arousal reactions may prove suitable for monitoring intra-anaesthetic nociception and its modulation. PMID- 7488486 TI - Monitoring lung compliance and end-tidal oxygen content for the detection of venous air embolism. AB - Venous air embolism (VAE) is a recognized complication of surgery performed with the patient in the sitting position, but it occurs also during other operations. We report two cases of VAE, associated with a notable decrease in dynamic lung compliance, detected by side-stream spirometry. Based on these cases, an experiment with 10 pigs was designed to evaluate the usefulness of side-stream spirometry in the diagnosis of VAE. Three doses of air (0.5, 1.0 and 2.0 ml kg-1) were injected via the proximal part of a 5- French gauge pulmonary artery catheter. Only the largest dose was followed by haemodynamic deterioration. Significant increases in end-tidal oxygen content and decreases in dynamic lung compliance were detected with all doses of air together with conventional signs of VAE, that is increases in pulmonary artery pressures and arterial carbon dioxide tensions, and decreases in end-tidal concentration of carbon dioxide. We conclude that continuous monitoring of end-tidal oxygen concentration and side stream spirometry offers valuable supplements to other monitoring techniques in the detection of VAE. PMID- 7488487 TI - Propofol-induced alterations in the microcirculation of hamster striated muscle. AB - To investigate the effects of propofol on small vessels, we have measured changes in diameter and blood flow in microcirculatory venules and arterioles. Studies were carried out in the dorsal skinfold chamber of hamsters by intravital fluorescence microscopy. A bolus injection of propofol 25 mg kg-1 dilated small and collecting venules by a mean value of 18% and arterioles by 13%. Blood flow in venules increased by up to 69%. Intralipid dilated arterioles and post capillary venules by 26% and 30%, respectively. After 4 h of continuous infusion (0.2-0.8 mg kg-1), only blood flow in post-capillary venules increased (66% propofol and 92% Intralipid). Post-capillary and collecting venules dilated in both groups. Therefore, propofol and Intralipid induced venodilatation and enhanced blood flow after bolus administration. After 4 h, despite dilatation in both groups, only post-capillary venules showed enhanced blood flow. These observations suggest redistribution of blood flow after prolonged administration of propofol. PMID- 7488488 TI - Effects of propofol on isolated rabbit mesenteric arteries and veins. AB - We have investigated the effect of propofol on isolated rabbit mesenteric arteries and veins. Isometric tension was measured in rings of arteries (with or without endothelium) or veins in organ chambers. The preparation was stimulated with noradrenaline 10(-6) mol litre-1, K+ 50 mmol litre-1 and caffeine 20 mmol litre-1 in the presence or absence of propofol. Propofol potentiated noradrenaline-induced contractions at lower concentrations (3 x 10(-5) mol litre 1) and attenuated them at greater concentrations (10(-4) and 3 x 10(-4) mol litre 1) in arteries with endothelium. Propofol inhibited noradrenaline-induced contractions in arteries without endothelium. In contrast, propofol produced venodilatation in a concentration-dependent manner (10(-5) to 3 x 10(-4) mol litre-1) of significantly greater magnitude than that in arteries. Propofol inhibited K+-induced contraction of both arteries and veins. It decreased the relaxation induced by acetylcholine (3 x 10(-8), 10(-7) and 3 x 10(-7) mol litre 1) of noradrenaline-induced contractions of arteries. Propofol did not affect caffeine-induced contractions after pretreatment with increased Ca2+. We conclude that propofol has a more potent vasodilator effect on veins than on arteries. Vasoconstriction induced by propofol may be associated with inhibition of endothelium-derived relaxing factor, whereas vasodilatation induced by propofol may be associated with block of voltage-gated influxes of extracellular Ca2+. PMID- 7488489 TI - Difference between in vivo and in vitro effects of propofol on defluorination and metabolic activities of hamster hepatic cytochrome P450-dependent mono oxygenases. AB - We have compared the in vivo and in vitro effects of propofol on cytochrome P450 dependent monooxygenase activities in hamster liver microsomes. Propofol (Diprivan) 10 mg/100 g body weight was injected i.p. twice a day for 2 weeks to induce cytochrome P450 enzymes. Liver microsomes were prepared by differential centrifugation. Metabolism of the cytochrome P450-dependent mono-oxygenase system was evaluated by measuring aniline hydroxylation, benzphetamine demethylation and benzo(a)pyrene hydroxylation. Defluorination of enflurane was assayed by detecting free fluoride metabolites. At similar concentrations as in the in vivo group, propofol in vitro exhibited concentration-dependent inhibition of metabolism of benzphetamine and benzo(a)pyrene. Aniline hydroxylation and defluorination of enflurane were inhibited to 78% of control with propofol 0.25 mmol litre-1. In propofol-treated hamsters, there was only minimal inhibitory or inductive effects on either mono-oxygenase activities or capacity for defluorination. This difference between the in vitro and in vivo effects of propofol on cytochrome P450 mono-oxygenase activities emphasizes the need for care when comparing in vitro and clinical data. PMID- 7488490 TI - Alterations of inositol polyphosphates in skeletal muscle during porcine malignant hyperthermia. AB - Malignant hyperthermia (MH) may result from increased intracellular calcium concentrations. Increased 1,4,5-IP3 concentrations could mediate this increase in Ca2+. In this study we measured inositol polyphosphates in selectively bred MH susceptible (MHS) and MH non-susceptible (MHN) swine. MH crisis was induced by halothane challenge, and dantrolene was administered in order to measure inositol polyphosphates after MH reversal. Muscle biopsies of skeletal muscles of the hind limbs were obtained in random order and inositol polyphosphates determined by high pressure liquid chromatography using a metal dye detection method. Inositol polyphosphates were determined in three groups: (1) MHS vs MHN basal, (2) during MH crisis induced by halothane and (3) following treatment with dantrolene after halothane challenge. Clinical variables (P(_)VO2, P(-)VCO2, PE'CO2 and pH) indicated that MH was readily induced in MHS swine. Basal concentrations of all inositol polyphosphates were higher in MHS swine compared with MHN swine. After halothane challenge, 1,3,4-IP3, 1,3,4,6-IP4 and 1,3,4,5-IP4 concentrations increased in MHS animals compared with the respective baseline values, whereas no changes in MHN animals could be detected. Dantrolene administration decreased inositol polyphosphate concentrations in MHS swine. MHN swine showed no changes in inositol polyphosphates after dantrolene. These findings indicate that inositol polyphosphates may be involved in metabolic changes after triggering and treatment of MH. PMID- 7488491 TI - Virtual reality in anaesthesia. PMID- 7488492 TI - Use of the abbreviated mental test to detect postoperative delirium in elderly people. AB - We investigated the validity of the abbreviated mental test (AMT) as a guide to the diagnosis of delirium in 100 patients aged more than 65 yr. Patients were assessed using the AMT on the day before and on the third day after operation. Fifteen patients were delirious on the third postoperative day; 10 of 43 patients undergoing orthopaedic surgery and five of 57 patients undergoing non-orthopaedic surgery. Delirium developed in four of 16 patients with a preoperative AMT score less than 8 and in 11 of 84 patients with a preoperative AMT score of 8 or more. Patients who developed delirium had a greater decline in AMT score (mean 2.7 (SD 0.9)) than patients who did not develop delirium (0.7 (1.0)) (P < 0.001). The sensitivity and specificity of a decline in AMT score of 2 or more points after surgery for diagnosis of postoperative delirium were 93% and 84%, respectively. PMID- 7488493 TI - Use of i.m. temperature probes during cardiopulmonary bypass in humans. AB - I.m. temperature probes were used in humans during and after hypothermic cardiopulmonary bypass (CPB). After rewarming to apparent normothermia of the body core, we found that the temperature of the vastus lateralis muscle remained well below the nasopharyngeal temperature at the time of weaning from bypass. This reduced muscle temperature represents a considerable deficit in total body heat which may account for a large proportion of the afterdrop in temperature which occurs in the postoperative period. There was no morbidity associated with the use of the i.m. probes which could be incorporated into standard clinical practice. PMID- 7488494 TI - Anaesthetic management of acute puerperal uterine inversion. AB - We identified 18 cases of acute puerperal uterine inversion after vaginal delivery out of 65,581 deliveries in two university-affiliated hospitals (an incidence of 1:3643). In all cases manual repositioning of the uterus without tocolysis was attempted. In four cases this was successful. Of the 14 patients requiring tocolysis, a single dose of terbutaline 0.25 mg i.v. was tried in eight and was successful in five cases (success rate of 63%), while in three patients general anaesthesia was required. In six other patients, general anaesthesia was chosen as the initial management. There was no associated maternal mortality or morbidity with the exception of one case of acute postpartum endometritis. We suggest that when tocolysis is required, terbutaline should be used first because of its rapid onset of action, short duration, simplicity, availability in the labour suite and familiarity to the obstetrician, and general anaesthesia, with its inherent dangers, if possible, should be avoided. PMID- 7488495 TI - Double lung transplantation without cardiopulmonary bypass: arterial to end-tidal carbon dioxide partial pressure differences. AB - Bilateral lung transplantation without cardiopulmonary bypass consists of two sequential single lung transplantations. Variations in ventilatory status during the procedure led us to study the (PaCO2-PE'CO2) gradient to see if PE'CO2 might reflect PaCO2. The gradient was studied in 14 patients at six times during operation. (PaCO2-PE'CO2) (kPa) was mean 1.97 (SD 0.7) after induction, 3.2 (1.4) during single lung ventilation, 1.9 (1.1) after clamping of the contralateral pulmonary artery, 2.96 (1.6) after ventilation and vascularization of the first transplant and the remaining native lung, 0.99 (0.8) during single lung ventilation with the first transplant and 1.3 (0.8) during ventilation of both transplants. With ventilation by the allograft lung(s) alone, the small (PaCO2 PE'CO2) value demonstrated improvement in ventilatory status, enabled PaCO2 to be assessed by PE'CO2 and demonstrated efficiency of the grafts. PMID- 7488496 TI - Early detection and treatment of myocardial ischaemia after operation using continual ambulatory arterial pressure monitoring and ECG ST segment analysis. AB - We report a case in which the use of continual ambulatory arterial pressure monitoring and ECG ST-segment analysis allowed early detection and treatment of myocardial ischaemia in the postoperative period. We believe that this case illustrates the potential value of ambulatory monitoring in the early postoperative period in high-risk patients. PMID- 7488497 TI - Transoesophageal echocardiographic examination of a patient with venacaval and pericardial tears after blunt chest trauma. AB - A 58-yr-old male was admitted with blunt thoracic and abdominal trauma. Transoesophageal echocardiography (TEE) was performed acutely to determine the cause of intrathoracic haemorrhage. We found atrial septal haematoma and tear, which have not been described previously, and which may be useful indicators of major intrathoracic venous tears which are always difficult to diagnose. Although the outcome was not altered in this case, we feel that TEE is a useful adjunct in the diagnosis of acute thoracic trauma. PMID- 7488498 TI - Mivacurium chloride and myotonic dystrophy. AB - We describe the successful use of the short-acting, non-depolarizing neuromuscular blocking agent, mivacurium, in a patient with myotonic dystrophy. Increased sensitivity to mivacurium was demonstrated using train-of-four monitoring, with a single dose of mivacurium providing adequate block for 90 min of surgery. Spontaneous recovery appeared prolonged with a recovery index (25-75% T1) of 10 min and a recovery time (5-95% T1) of 30 min. The use of reversal agents and anticholinergic agents was avoided. PMID- 7488499 TI - Management of puffer fish poisoning. PMID- 7488500 TI - Tramadol. PMID- 7488501 TI - Tramadol. PMID- 7488502 TI - Post traumatic stress disorders. PMID- 7488503 TI - Maximum doses of local anaesthetics. PMID- 7488504 TI - In defence of the Arkive. PMID- 7488506 TI - Convective warming after hypothermic cardiopulmonary bypass. PMID- 7488505 TI - Use of the lighted stylet for tracheal intubation via the laryngeal mask airway. PMID- 7488507 TI - Red blood cell substitutes. PMID- 7488508 TI - Incidents and critical incidents. PMID- 7488509 TI - Cutaneous heat loss during anaesthesia. PMID- 7488510 TI - Pharmacokinetics of ketamine enantiomers. PMID- 7488511 TI - Gastric tonometry. PMID- 7488512 TI - Effect of i.v. diamorphine on regression of spinal block. PMID- 7488513 TI - CD15 and HKN-1 reveal cerebellar compartments with a complex overlap. AB - The CD15-epitope in the cerebellar cortex is expressed by Bergmann glial fibers which are organized in symmetrical iterative stripes. Their arrangement appears complementary to the pattern described for the organization of compartments demonstrated by any other marker used so far. HNK-1 however, reveals stripes that are not complementary to the general pattern for the organization of compartments. PMID- 7488514 TI - Expression of CD15 in a subset of dorsal root ganglion cells during the chick embryonic development. AB - We have investigated the distribution of CD15 immunoreactivity (IR) in chick sensory neurons during the embryonic development. IR first appeared around embryonic day 6 (E6) preferentially in the lateroventral part of the dorsal root ganglion (drg). Here it was located in the cytoplasm of the ganglion cells. Starting at E11, CD15 positive cells have been detected also in the dorsomedial part of the drug. Towards the end of embryonic development, these immunoreactive (ir) cells were evenly distributed within the drg. Around E 16 nerve fibres could also be found, expressing the CD15 epitope. These fibres run both in centripetal and centrifugal direction, and could be followed throughout peripheral nerves. IR was confined to peripheral nerves of the skin, where after its subepidermal arborisation fine nerve terminals entered the basal stratum of the epidermis or the dermal bulbous part of feathers. Until hatching no staining in visceral structures was detected. Our results suggest that CD15 is restricted to a subset of drg neurons innervating somatosensory targets. A role for the CD15 epitope for myelination is discussed. PMID- 7488515 TI - Age-related expression of the CD15 (3-fucosyl-N-acetyl-lactosamine) epitope in the monkey (Cercopithecus aethiops aethiops L.) lateral geniculate nucleus. AB - The profile of the CD15 epitope-expression was studied in the lateral geniculate nucleus (LGN) of a monkey species (Cercopithecus aethiops aethiops L.) during the peri- and postnatal development. Intense labeling of the neuropil and of astroglial cells was detected. Morphological examination of the spatio-temporal appearance of the CD15-expression showed different consecutive patterns of expression: expression of the CD15 epitope was high one week prenatally and during the first postnatal weeks. During this perinatal period it was mainly expressed in the interlaminar zone of the magnocellular portion of the LGN. This was followed by a time period when immunoreactivity gradually decreased to become almost absent by approximately 10 weeks of age. Toward adulthood a different pattern of immunoreactivity occurred, revealing a pattern of lamination that was attributable to CD15 positive astrocytes, most prominent in the cellular layers 1 and 2. They were later on seen spread out through the entire LGN such that the LGN of adult animals was entirely filled with positive material. From the temporal correlation of CD15-expression and LGN histogenesis it is concluded that the high level of CD15, that is observed in the neuropil during the perinatal period, matches with high synaptic plasticity within the visual system; in contrast, low levels of CD15-expression correlate with synaptic reorganization processes. PMID- 7488516 TI - Expression of the carbohydrate epitope 3-fucosyl-N-acetyl-lactosamine (CD 15) in the adult guinea pig inner ear. AB - The expression of the CD 15 (3-fucosyl-N-acetyl-lactosamine) epitope was immunohistochemically studied on paraffin sections of adult guinea pig inner ears. Two regions of the inner ear expressed the epitope for CD 15: the tectorial membrane of the cochlea and the endolymphatic sac. The upper part of the main body of the tectorial membrane was deeply stained. In the rugosal and distal part of the endolymphatic sac several unevenly distributed cells showed strong intra- and extracellular localization of the CD15 epitope. The CD15 epitope is associated with a transduction structure (tectorial membrane) and with a "volume regulating" compartment (endolymphatic sac) and may be involved in the maintenance of the structural integrity of both. PMID- 7488517 TI - Simultaneous demonstration of CD15 and alkaline phosphatase activity in cryostat sections of rat fetuses: a detailed technical description for the developing brain. AB - CD15 and alkaline phosphatase are significant markers of the developing central nervous system. CD15 is known for its interaction in cell-cell contacts, while the presence of alkaline phosphatase is related to the formation of neuronal connections. This paper describes a combined immunocytochemical and enzyme histochemical technique to demonstrate both embryonic markers in the same cryostat section. The monoclonal antibody 3B9, which was used for the detection of CD15 recognizes the immunodominant carbohydrate structure alpha-1,3-Fucosyl-N Acetyl-Lactosamine also known as X, LewisX, FAL or SSEA-1. This antigen dissolves and is easily rinsed out of the section. The procedure starts off with the detection of CD15. The results of the immunocytochemical procedure is a reaction product which is stable at a high pH, preventing loss of the immunocytochemical reaction product during the subsequent alkaline phosphatase detection. The other embryonic marker, alkaline phosphatase, can only be demonstrated enzymehistochemically if the enzyme is still active. The immunocytochemical localization procedure decided for, does not inactivate alkaline phosphatase totally. From the colorimetrical and histochemical alkaline phosphatase determinations it was concluded that the residual alkaline phosphatase activity detected with this technique, could be intensified by adding Mg(2+)-ions to both the colorimetrical and histochemical incubation media. PMID- 7488518 TI - Spotting hot microwave problems. PMID- 7488519 TI - N-[18F]-fluoroalkylation of noraporphines: microwave versus thermal treatment. AB - A comparable study of microwave versus thermal heating is described for the N [18F]-fluoroalkylation of noraporphines. As compared to thermal treatment, different products were obtained during microwave treatment. Thermal treatment resulted in the loss of the protection of the catechol functionality of the noraporphines (O-deacylation), whereas during microwave treatment N-[18F] fluoroalkylation was observed. The results described in this report might suggest that the influence of microwaves on chemical transformations is not exclusively thermal. PMID- 7488520 TI - Handling of large brain specimens: the use of microwaves. A review. PMID- 7488521 TI - Combining microwave stabilization and microwave-stimulated fixation of brain tissue with microwave-stimulated staining. AB - Microwave stabilization of rat brain tissue using saline was compared with microwave-stimulated fixation using either formaldehyde or Kryofix. Sections were stained according to the microwave-adapted Nissl, Kluver-Barrera, and Bodian methods. Depending on the preparative choice the microwave staining method had to be adjusted. Optimal combinations of preparative techniques and staining methods are feasible. In most cases the microwave saline-stabilized and microwave Kryofix treated sections were better suited for application in neurohistology than the microwave formaldehyde-treated sections for light microscopy. PMID- 7488522 TI - Introduction: workshop summary of the CD15 monoclonal antibody panel from the Fifth International Workshop on Leukocyte Antigens. AB - CD15 is expressed on a wide variety of tumor cells including myeloid leukemia, breast, colorectal, lung cancer cells. It is probable that the core proteins and lipids differ on the different cell types since there is marked variability in the binding of the mAb within and among the different cell types. Previously, there had only been a single mAb of the IgG class directed to CD15. The present study demonstrates the existence of one more IgG mAb (an IgG3). Unfortunately, a third mAb submitted as an IgG1, MA14, did not appear to be an IgG in our studies and a fourth mAb, MA9, submitted as an IgG1 did not have significant binding activity and also could not be confirmed as anti-CD15. An interesting finding was that all of the mAb could mediate complement-dependent cytotoxicity with rabbit serum. However, the two IgG3 mAb, MA63 (MCS-1) and MA88 (7C3), could not mediate lysis with human complement, while all of the IgM mAb could do so. It is not clear why this difference exists. CD15 is known to be expressed on mature myeloid cells in the hematopoietic lineage. The expression of CD15 on progenitor cells has been mapped by the indirect method of C'-dependent lysis and measurement of colony-forming cells. These studies have estimated that about 50% of normal CFU GM express CD15. In the present workshop, we examined the expression of CD15 on a quantifiable cell population, the CD34 positive population.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7488523 TI - Comparison of resistance of fungal spores to gamma and electron beam radiation. AB - The irradiation sensitivity of fungal spores to either gamma or electron beam irradiation was evaluated in distilled water. The D10 (the dose required to reduce the initial population by 90%) gamma values ranged from 0.236 to 0.416 kGy and from 0.209 to 0.319 kGy for Penicillium and Aspergillus species, respectively. The D10 electron beam values ranged from 0.194 to 0.341 and from 0.198 to 0.243 kGy for Penicillium and Aspergillus species, respectively. Of the aspergilli species evaluated, only half exhibited significantly (P < 0.05) greater sensitivity to the electron beam treatment compared to gamma irradiation. Four of the six penicillia species evaluated also exhibited significantly (P < 0.05) higher sensitivities to electron beam treatment. PMID- 7488524 TI - Attributes of microbial associations of meat growing as xenic batch cultures in a meat juice at 4 degrees C. AB - Strains of Gram-positive (Carnobacterium piscicola, Leuconostoc mesenteroides subsp. mesenteroides, Brochothrix thermosphacta) and Gram-negative (Pseudomonas fragi and Hafnia alvei) bacteria isolated from minced lamb packaged under a modified atmosphere were cultivated in a meat (lamb) juice at 4 degrees C. Carbohydrates were catabolised in the order glucose > glucose 6-phosphate during the development of the population. Under an atmosphere enriched with carbon dioxide the Gram-negative portion of the population was suppressed during the exponential phase but H. alvei became the dominant organism towards the end of a protracted stationary phase of growth. With the aerobic atmosphere P. fragi catabolised creatine and became the dominant species in the stationary phase. The inability of C. piscicola to catabolise glucose 6-phosphate was reflected in its population being smaller than those of the other Gram-positive organisms (C. piscicola < L. mesenteroides subsp. mesenteroides < B. thermosphacta). During the stationary phase of growth, indigenous L-lactic acid and the D-isomer produced by leuconostocs were oxidised to acetic acid by the Gram-positive flora under an atmosphere enriched with carbon dioxide. These oxidations, which occurred after depletion of glucose, were supported by the oxygen in the system. D-Lactic and acetic acid appeared to be possible parameters for the estimation of the microbiological quality of packaged meat. PMID- 7488525 TI - The isolation of Campylobacter jejuni from contaminated surfaces and its survival in diluents. AB - The isolation rates of campylobacters from contaminated surfaces were improved if swabs were placed directly into selective media rather than being stored in diluents before culture. Storage in diluents resulted in a loss of viability and the remaining viable campylobacter cells were often sub-lethally injured which sensitised them to selective agents in culture media and reduced isolation rates. Campylobacter jejuni, suspended in small drops of blood, was capable of prolonged survival on work surfaces if the drops remained liquid but the bacterium died rapidly once drops had dried. PMID- 7488526 TI - Modelling of microbial activity and prediction of shelf life for packed fresh fish. AB - Prediction of shelf life based on growth of specific spoilage organisms (SSO) in model substrates was studied. The effect of CO2 on the growth kinetics for Photobacterium phosphoreum and Shewanella putrefaciens was quantified and modelled. Results showed that microbial spoilage of packed cod stored with various concentrations of CO2 was accurately predicted from the effect of CO2 on P. phosphoreum grown in model substrates. The short shelf life extensions previously reported for packed cod therefore can be explained by the high CO2 resistance of this Gram negative organism. S. putrefaciens was very sensitive to CO2 and growth rates could not be related to the shelf life of packed cod. Growth curves without lag phases were found for all concentrations of CO2 and for both the microorganisms studied. For the fitting of these growth curves the log transformed Logistic models were selected after comparison with the 'modified Gompertz' models and with the model of Baranyi et al. (1993). The effect of CO2 on mu max was well described by a 2 parameter square root model. Validation of kinetic models by comparison of shelf life predictions with shelf life determined by sensory evaluations in product experiments was preferred for comparison of microbial growth rates determined in product and model system experiments. Kinetic modelling was found to be valuable for both evaluation and prediction of microbial fish spoilage and an iterative approach for development of kinetic shelf life models was suggested. PMID- 7488527 TI - Qualitative and quantitative characterization of spoilage bacteria from packed fish. AB - The large cells recently suggested to be responsible for spoilage of packed cod, have been identified as Photobacterium phosphoreum. The spoilage activity of these cells, of Shewanella putrefaciens and of other microorganisms isolated form spoiled packed cod has been studied. Both qualitative and quantitative tests were used for characterization of the microbial spoilage activity. The importance of the different groups of microorganisms was evaluated by comparison of microbial spoilage activity determined in model substrates and in product experiments. The yield factor for production of trimethylamine (YTMA/CFU) and the cell concentration determined at the time of off-odour detection were used as quantitative measurements of microbial spoilage activity. On average cells of P. phosphoreum produced 30 times more TMA than cells of S. putrefaciens, YTMA/CFU of the two organisms were 10(-8.0) mg-N TMA/cfu and 10(-9.5) mg-N TMA/cfu, respectively. With these yield factors the level of TMA found in spoiled packed cod (30 mg-N TMA/100g) corresponds to about 10(7) cfu/g of P. phosphoreum and to 10(8)-10(9) cfu/g of S. putrefaciens. 10(7) cfu/g of P. phosphoreum were actually found in spoiled packed cod suggesting this organism could be responsible for spoilage. High cell concentrations of more than 10(8) cfu/g of S. putrefaciens were required for production of detectable off-odours and is was concluded that this organism is without importance for spoilage of packed cod. PMID- 7488532 TI - Comparison of the incidence of Listeria on equipment versus environmental sites within dairy processing plants. AB - This study was undertaken to compare the incidence of Listeria contamination of processing equipment with that of the general dairy processing environment. A total of 378 sponge samples obtained from 21 dairy plants were analyzed for Listeria using three different enrichment media. Use of extended microbiological analysis allowed us to identify 26 Listeria positive sites which would have not been identified had a single test format been employed. Eighty (80) of 378 sites (21.2%) were identified as Listeria positive. Listeria innocua was isolated from 59 of the 80 (73.8%) positive samples, L. monocytogenes was identified in 35 (43.8%) of the positive samples, and L. seeligeri was isolated from 5 (6.3%) of the Listeria positive samples. Positive equipment samples were obtained from 6 of the 21 (28.6%) plants and 19 of the 21 (90.5%) plants had positive environmental sites. Seventeen of the 215 (7.9%) samples from equipment were positive for Listeria species. Eleven of these sites, including 3 holding tanks, 2 table tops, 3 conveyor/chain systems, a pasta filata wheel, a pint milk filler and a brine pre-filter machine, were positive for L. monocytogenes. Nineteen of the 21 (90.5%) plants had positive environmental sites. Sixty-three of the 163 (41.1%) samples from environmental sites were Listeria positive and 24 were positive for L. monocytogenes. Two-tailed student t-test analysis of the mean frequencies indicated that the level of contamination was significantly higher (p < 0.001) in 'environmental' (49.7%) as opposed to 'equipment' samples (7.0%). Our study indicates that environmental contamination with Listeria does not necessarily translate into contamination of equipment within the same plant, and that greater emphasis needs to be placed on the cleaning and sanitizing of the plant environment. PMID- 7488530 TI - Contamination profiles and characterisation of Bacillus species in wheat bread and raw materials for bread production. AB - The Bacillus counts in white and wholemeal wheat loaves produced without preservatives or sour dough were consistently 10(6) cfu/g after two days of storage at ambient summer temperatures (25-30 degree C). Identified species were B. subtilis (70%), B. licheniformis (24%), B. pumilus (2%) and B. cereus (2%). The dominance of B. subtilis in bread could be explained by the higher resistance to heat of this species as determined by inoculation studies. Among 14 species isolated from retail bread and wheat grains, B. subtilis was the only species associated with ropiness. Samples of raw materials, particularly bran, seeds and oat products, contained low levels (10(0) - 10(2) cfu/g) of Bacillus spores, surviving a heat treatment (100 degree C, 10 min) corresponding to a baking process. Even low spore levels in raw materials with the frequently isolated species, B. licheniformis (49%) and B. subtilis (10%), resulted in 10(7) Bacillus per g bread crumb in two days as determined by test bakings. The results indicate a need for controlling growth of Bacillus in bread. PMID- 7488528 TI - The use of a PCR-generated invA probe for the detection of Salmonella spp. in artificially and naturally contaminated foods. AB - Part of the invasion A gene (invA) of slamonellae (Rahn et al., 1992) was amplified and labelled simultaneously with digoxigenin by the polymerase chain reaction (PCR). This was used as gene probe for a colony hybridization assay which included nitrocellulose filter incubation on modified Rambach agar. 312 Salmonella and 268 non-Salmonella strains were hybridized with the invA probe. No false-negative or false-positive results were obtained. In 11 beef samples, which had been contaminated artificially with Salmonella, the test strain was recovered quantitatively with the invA probe. Salmonellae could be detected in 29 samples of 104 further foods of animal origin by means of the gene probe assay in contrast to 27 samples which were positive by the standard method. The invA probe assay allows for the quantitative estimation of Salmonella in fresh meat samples within 48 h. However, with frozen samples a pre-enrichment step is necessary. PMID- 7488529 TI - Differentiation of dextran-producing Leuconostoc strains from fermented rice cake (puto) using pulsed-field gel electrophoresis. AB - Lactic acid bacteria were isolated from puto, a fermented rice cake consumed as a breakfast and snack food in the Philippines. The microflora was dominated by dextran-producing leuconostocs, and these were differentiated into four groups using pulsed-field gel electrophoresis of restriction enzyme digested chromosomal DNA, in conjunction with taxonomic tests. The four groups corresponded to the species Leuconostoc mesenteroides subsp. mesenteroides, Leuconostoc pseudomesenteroides, Leuconostoc citreum and Leuconostoc fallax. Several strains showed an unusual clumping phenotype, and two of these were capable of inhibiting other strains of lactic acid bacteria. PMID- 7488531 TI - Short-chain fatty acids modulate growth of lactobacilli in mixed culture fermentations with propionibacteria. AB - Three strains of Lactobacillus (L. helveticus ATCC 15009 and CRL 581, and L casei LC 3) were paired with three strains of Propionibacterium (P. freudenreichii AP8, P. freudenreichii GP6 and P. acidipropionici CRL 756) and grown in individual and mixed cultures in a complex medium. Bacterial growth, carbohydrate consumption, and production of acids was determined and compared after mono and binary culture. Propionibacterium strains were inhibited in mixed cultures that rapidly reached low pH values, and stimulated in those with slow pH reduction. In several pairs Lactobacillus strains were stimulated by mixed cultivation, while in others they were unaffected or inhibited. Lactic acid reduction by Propionibacterium consumption was not always enough to produce a stimulatory effect on growth of lactobacilli. The behaviour of Lactobacillus strains in individual cultures in media with propionic acid was similar to that of mixed cultures with Propionibacterium. As propionic acid concentration increased in the medium and glucose was consumed, both individual and mixed cultures of lactobacilli showed inhibition of the growth and increase of the fermentation activity. PMID- 7488533 TI - Dermatologic surgery and AIDS. Introduction. PMID- 7488534 TI - The epidemiology of HIV/AIDS. PMID- 7488535 TI - Transmission of HIV infection. PMID- 7488536 TI - Risk of acquiring AIDS from salivary exchange through cardiopulmonary resuscitation courses and mouth-to-mouth resuscitation. AB - In summary, the risk of transmission of HIV and other infectious diseases by saliva during CPR training practice is extremely low because of low infectious virus titers and properties of saliva that inhibit HIV. However, it is necessary to perform decontaminations of mannequins, by application of a suitable disinfectant and by a mechanical wipe-down with a sponge, to cleanse the external buccal area of the mannequin after contact with each CPR trainee. For health care and public safety professionals training and performance of MTM ventilation during CPR should be carried out with barrier devices such as the bag-valve-mask or face shield. Guidelines and standards of the AHA, American Red Cross, and the CDC for prevention of infection during CPR and emergency cardiac care are more fully available elsewhere. If the recommended procedures are followed, the risk of acquiring HIV from saliva during MTM should be extremely low. PMID- 7488537 TI - HIV infection in healthcare workers: risk for infection and methods of prevention. PMID- 7488538 TI - The potential role of simple dermatologic surgery techniques in transmitting HIV infection. PMID- 7488539 TI - Infection control precautions for surgical personnel in the surgery unit and for laboratory personnel in the Mohs surgery unit laboratory areas. PMID- 7488540 TI - Management of occupational exposure to HIV. PMID- 7488541 TI - Dermatologic surgery personnel with AIDS. PMID- 7488542 TI - Sterilization and disinfection of instruments and materials in the surgery unit and laboratory areas. PMID- 7488543 TI - Managing the infectious waste of the surgery unit and laboratory. PMID- 7488544 TI - Patient rights and physician responsibility: four problems in AIDS care. PMID- 7488545 TI - Animal models of inflammatory bowel disease. PMID- 7488546 TI - The cochlear lesion in experimental bacterial meningitis of the rabbit. AB - Sensorineural hearing loss was studied in a rabbit model of experimental bacterial meningitis using electrophysiological and ultrastructural techniques. Hearing impairment was monitored by auditory brain-stem evoked responses (ABERs) and concomitant structural lesions were identified by both transmission (TEM) and scanning (SEM) electron microscopy. Meningitis was induced by intra-cerebrospinal fluid injection of either Escherichia coli (strain 2073 and type K-12) or Haemophilus influenzae type b. Auditory loss of approximately equal to 10 dB occurred in all rabbits by about 10 hours post infection and progressed in severity until by 20 h following infection, hearing losses up to and > 60 dB were obtained. At levels of hearing loss < 20 dB ultrastructural damage to the organ of Corti was barely detectable. With greater levels of hearing loss, patchy structural damage to hair cells, synaptic nerve terminals, supporting cells and inner spiral sulcus cells and cells of the stria vascularis was clearly evident. Bacteria were found in scala tympani, the basilar membrane, the organ of Corti, scala media, the spiral ligament and at the margin of the stria vascularis. Evidence of bleeding was found in some cochleas; erythrocytes were found in scala tympani, scala media, amongst hair cells and beneath the tectorial membrane. The results show that hearing loss is associated with bacterial invasion and damage to the organ of Corti and that the cause of hearing loss is likely to result from multiple lesions within the cochlea. Lesions to sensory cells almost certainly will produce permanent hearing loss. Lesions to supporting cells, nerve terminals and to stria vascularis may well produce only temporary hearing loss. PMID- 7488547 TI - Differential effects of interferon-gamma and -beta on fatty acid turnover, lipid bilayer fluidity and TNF-alpha release in murine macrophage J774.2 cells. AB - The effects of interferon (IFN)-gamma and IFN-beta on the incorporation of 14C linoleic acid into J774.2 cell membrane phospholipids were examined. Interferon gamma induced a statistically significant increase in incorporation of 14C linoleic acid into all the major phospholipid classes. In contrast, IFN-beta induced a slightly reduced incorporation of this fatty acid into the phospholipids. Neither IFN-gamma nor IFN-beta had any effect on the incorporation of the saturated fatty acid 14C-stearic acid into the cellular phospholipids. Interferon-gamma had no effect on the metabolism of 14C-linoleic acid in the fibroblast cell line L929. Macrophage membrane fluidity was assessed by spin label ESR spectroscopy after incubation with either IFN-gamma or IFN-beta. Interferon-gamma significantly increased membrane fluidity whereas IFN-beta significantly decreased the fluidity. The findings of this study reveal that IFN gamma might act on the enzymes controlling the labelling of the sn2 position of phospholipids (linoleic acid) but not the sn1 position (stearic acid), and this increases the polyunsaturated fatty acid content of macrophage membranes. This increase in polyunsaturation is reflected in the increased membrane fluidity. We also conclude that IFN-beta and IFN-gamma have different mechanisms of action on macrophage membrane lipid metabolism. PMID- 7488548 TI - Scanning electron microscopic changes in morphology of pulmonary endothelium in rat lung isografts following hypothermic ischaemic storage and transplantation. AB - Endothelial monolayer integrity is a critical factor limiting vascular permeability of solid organs in transplantation. Several in vitro, ex vivo and in vivo studies suggest that damage to endothelial cells (EC) due to hypothermia and ischaemia-reperfusion injury causes morphological and functional damage to the endothelium leading to parenchymal oedema and haemorrhage. Aiming to study morphological changes to arterial pulmonary EC subjected to transplantation procedures, random scanning electron micrographs of vascular endothelium of rat lungs were taken. Forty-eight rat lungs were hypothermically stored for 48 or 72 hours in two different preservation solutions and studied either at the end of the cold storage period, or 5 min, 24 h or 4 weeks following transplantation. After 5 minutes of revascularization, micrographs showed EC shape variations, bleb formation and cell retraction with intercellular gap formation. Twenty-four hours after transplantation loss of monolayer continuity was widely extended. Four weeks of revascularization resulted in either well preserved specimens with nearly normal endothelium, or badly preserved arteries with fibrotic degeneration of the luminal vessel wall. The morphological disruptions found in this study help to explain the alterations in permeability control and vascular dysfunction observed in lung transplantation. PMID- 7488549 TI - The effect of Kupffer cell elimination on ethanol-induced liver damage in mice. AB - C57BL/10 mice develop inflammatory and necrotic changes in the liver, as well as raised serum ALT activities, after 9 days of exposure to ethanol vapour. If mice were injected twice with liposomes containing dichloromethylene diphosphonate (DMDP), with an interval of 5 days between the injections, there was complete elimination of Kupffer cells (hepatic macrophages) for a 9-day period starting 1 day after the first injection. The inflammatory and necrotic changes were significantly reduced in mice injected with liposomes containing DMDP as compared to uninjected mice or mice injected with empty liposomes; serum ALT activities were also significantly reduced. No significant difference was seen in serum tumour necrosis factor-alpha levels between the different groups. Kupffer cells therefore play a significant role in the development of the liver damage resulting from exposure to ethanol. Acetaldehyde production by Kupffer cells is one way in which these cells can damage hepatocytes and further work needs to be done to investigate this and other mechanisms. PMID- 7488551 TI - Suppression of pannus-like extension of synovial cells by lipid-derivatized chondroitin sulphate: in vitro and in vivo studies using Escherichia coli-induced arthritic rabbits. AB - In rheumatoid arthritis, pannus formation resulting from synovial inflammation is a major factor in cartilage destruction. The ability of arthritic synovial cells to undergo pannus formation depends upon their initial adhesion to the partially deformed cartilage surfaces. Our recent studies using various lipid-derivatized glycosaminoglycans have revealed a preeminent inhibitory activity of phosphatidyl ethanol amine-derivatized chondroitin sulphate (CS-PE) toward cell-matrix adhesion. Here we evaluate whether CS-PE may protect articular cartilage from pannus extension in different in vitro and in vivo model systems using Escherichia coli 0:14-induced arthritis in rabbits and the articular cartilage explants, synovial tissues, and synovial cells obtained from them. These studies showed that CS-PE suppressed the in vivo pannus-like extension on cartilage surfaces, as well as the in vitro extension of the synovial cell layer on both CS PE treated culture plates and cartilage explants. The results suggest that native chondroitin sulphate proteoglycans in the surface of normal articular cartilage play an important role in protecting the tissues from pannus extension and that the CS-PE immobilized onto partially eroded cartilage can mimic the inhibitory action of native chondroitin sulphate proteoglycans. PMID- 7488550 TI - A new fibroblast growth stimulating activity from the human megakaryoblastic leukaemia cell line ELF-153: in vitro and in vivo findings. AB - Although the exact mechanism for the progression of myelofibrosis in acute megakaryoblastic leukaemia is unclear, certain humoral factors released from the proliferating megakaryoblasts that are unable to store these factors in their defective alpha-granules, including platelet derived growth factor (PDGF), fibroblast growth factors (FGF), platelet factor-4 (PF-4), transforming growth factor-beta (TGF-beta) and beta-thromboglobulin, could result in increased collagen synthesis by bone marrow fibroblasts. Recently, the human megakaryoblastic leukaemia cell line MEG-01 has been shown to produce both TGF beta and PF-4 which have enhanced the growth of bone marrow fibroblasts. Therefore, we have examined the presence of a fibroblast growth stimulating activity and the humoral factors that might be responsible for it in the supernatant of the human megakaryoblastic leukaemia cell line ELF-153 recently established in our laboratory from a patient with acute myelofibrosis. A new fibroblast growth stimulating activity has been identified in the supernatant of the ELF-153 human megakaryoblastic leukaemia cell line that is independent of the percentage of fetal calf serum in NRK-49F fibroblast agar clonogenic assays and is not due to any of the known fibroblast growth stimulating humoral factors including PDGF, epithelial growth factor, TGF-alpha or beta, tumour necrosis factor-alpha, interleukin-1, 2, 4 or 6, FGF, fibronectin, PF-4 and factor VIII AG. Also, in vivo, subcutaneous injection of ELF-153 megakaryoblastic leukaemia cells into nude mice formed, in three out of the five mice after 6 weeks, subcutaneous tumours with a very rigid texture whose histological examination revealed dense infiltration by blast cells and pronounced reticular fibrosis. Immunohistochemistry demonstrated exclusive deposition of collagen III in the extracellular matrix whereas laminin and collagen IV were absent.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7488552 TI - Evolution of endotoxin induced acute lung injury in the rat. AB - To clarify the evolution of acute lung injury induced by endotoxin, the progression of lung damage in 26 rats submitted to intratracheal instillation of 5 mg/kg body weight endotoxin was examined by blood gas analysis, computerized tomography, light and electron microscopy. Hypoxaemia, hypercapnia, acidosis and inhomogeneous bilateral infiltrates developed gradually within 48 hours. Monocytes appeared within blood capillaries and the instertitium by 12 hours after treatment, then migrated into alveoli and underwent progressive differentiation into macrophages by 24 hours after treatment. Granulocytes were found within blood capillaries at an early stage, but outside capillaries only at 48 hours. Hyperplasia of type II pneumocytes and hypertrophy of interstitial fibroblasts also occurred at 48 hours. These data suggest that the pathogenesis of endotoxin induced pulmonary injury proceeds through an early phase of granulocyte migration inside capillaries and monocyte extravasation, an intermediate phase of monocyte differentiation into macrophages inside alveoli and a late phase of diffuse infiltration of alveoli by newly differentiated macrophages and late-extravasated neutrophils. PMID- 7488553 TI - Gene therapy and HIV disease. PMID- 7488554 TI - Bacterial infections in HIV disease. PMID- 7488555 TI - Mycobacterium avium complex infection in AIDS. PMID- 7488558 TI - New developments in antiretroviral drug therapy for HIV-1 infections. PMID- 7488557 TI - HIV viral load quantification, HIV resistance, and antiretroviral therapy. AB - We are moving rapidly beyond a "black box" understanding of the pathogenesis of HIV. The sites of virus replication, the molecular regulation of virus production in the host, and the dynamics between productive virus infection and immunological and clinical events are areas of intense study using powerful new tools. The quantitation of virus load and genetic characterization of replicating virus has important implications for the development and evaluation of drugs and treatment strategies for HIV. As new compounds are introduced, their ability to reduce virus load in vivo has become a primary consideration in the decision to initiate large efficacy trials and may soon be used, in combination with other markers, in the licensing of new agents. In parallel, rapid molecular evaluation of virus from patients, targeting those who break through drug-induced suppression, provides an explanation for the failure of drugs to sustain an effect on virus load. This approach has compressed the process of drug evaluation and set the stage for the evaluation of complex combinations and sequences of drugs to maintain suppression of virus and prevent the development of drug resistance. The most controversial question for the next few years is whether the measurement of virus load or detection of drug resistance can be incorporated into the practice of medicine and the management of individual patients. There is evidence that changes in virus load are the most proximate markers of drug response and that detection of resistance mutations can predict clinical and immunological decline. However, the window of time between a change in load or the development of drug resistance and a decline in CD4 cells is relatively short. With dideoxynucleoside therapies, a CD4 cell decline follows a rise in virus load or development of resistance within 3-6 months. In early studies with protease inhibitors and nonnucleoside reverse transcriptase inhibitors, the development of resistance and a return to baseline of virus load may occur within 2-3 months, mirrored by a fall in CD4 cells. The challenge to investigators is how to best use these new tools to determine whether changes or additions in therapy, initiated on the basis of virological measurements, result in more effective management of disease. PMID- 7488556 TI - Pathophysiology and treatment of the AIDS wasting syndrome. PMID- 7488559 TI - Update on epidemiology of AIDS in Asia. PMID- 7488560 TI - Multiple opportunistic pathogen prophylaxis for patients with advanced HIV disease: an overview. PMID- 7488561 TI - Pathogenesis and management of HIV-associated drug hypersensitivity. PMID- 7488562 TI - Cytomegalovirus retinitis in AIDS: natural history, diagnosis, and treatment. AB - Cytomegalovirus retinitis is the most common intraocular infection and the leading cause of blindness in patients with AIDS. Diagnosis is made on clinical grounds; a variety of other infectious and neoplastic retinitides should be considered in the differential diagnosis. Treatment with intravenous ganciclovir or foscarnet has been proven effective, but late complications of relapse, viral resistance, retinal detachment, and drug toxicity remain problematic. Approved and investigational drugs for the treatment of CMV retinitis are limited by their virostatic, rather than virocidal, properties. Because the efficacies of ganciclovir and foscarnet are similar, the choice of therapy should be based on systemic considerations such as drug toxicity and patient survival. The cost, toxicity, and limited efficacy of currently available therapy of CMV retinitis make the need for an effective prophylactic drug even more important. PMID- 7488563 TI - Positive and negative symptoms in neuroleptic-free psychotic inpatients. AB - Andreasen's scales for the assessment of positive (SAPS) and negative (SANS) symptoms and the Brief Psychiatric Rating Scale (BPRS) were administered to a group of 70 neuroleptic-free psychotic inpatients. Individual ratings from the SAPS and SANS, together with the 18-item BPRS, were examined to identify clusters of symptoms. The findings, consistent with our previous studies using medicated patients, did not support a simple positive-negative dichotomy. Independent syndromes representing negative symptoms and thought disorder were apparent, although within the negative syndrome there were three related sub-syndromes of flat affect, alogia and social dysfunctions. Hallucinations and delusions did not form a homogeneous group of symptoms. Paranoia emerged as a distinct syndrome, and the remaining symptoms could be subdivided into hallucinations, grandiose delusions, and 'loss of boundary' delusions (e.g., thought broadcasting, mind reading). These syndromes, with the exception of loss of boundary delusions, which consisted of SAPS delusions alone, correspond to syndromes of psychosis identified by Lorr and his colleagues thirty years ago. It is concluded that the currently popular 'three syndrome' model does not adequately represent the diversity of psychotic symptoms. PMID- 7488565 TI - Scaffolded instruction and the remediation of Wisconsin Card Sorting Test deficits in chronic schizophrenia. AB - The purpose of this study was to explore the remediation of Wisconsin Card Sorting Test (WCST) deficits among highly chronic schizophrenic patients by employing an instructional technique known as scaffolding. The extent to which learning of the WCST generalized to a similar neuropsychological test, the Short Category Test (SCT) was examined. Forty-one chronic schizophrenic subjects drawn from in-patient and out-patient wards were given the WCST. Twenty-eight of the subjects who displayed the greatest impairment on the WCST were then administered the Brief Psychiatric Rating Scale (BPRS) and randomly assigned to groups that received three sessions of either scaffolded or didactic instruction. Post testing with the WCST occurred an average of 1.9 (SD = 1.3) and again 31.4 (SD = 3.9) days after instruction. Subjects who received scaffolded instruction improved significantly in the number of categories achieved when compared to the didactic group. The improvement was sustained a month later. A strong trend favoring the scaffolded group on the SCT suggested that generalization of learning had occurred. Significant and durable improvement of WCST performance in severely disabled schizophrenics is clearly possible with the instructional method employed. The increment in abstract reasoning may generalize to other tasks. These findings have significant implications for psycho-social and cognitive rehabilitation. PMID- 7488564 TI - Premorbid adjustment as a predictor of phenomenological and neurobiological indices in schizophrenia. AB - Previous studies have demonstrated relationships between poor premorbid adjustment and a variety of phenomenological and neurobiological indices in schizophrenic patients. Using the Modified Premorbid Adjustment Scale we re examined these relationships in a large sample (n = 131) of schizophrenic patients. Subjects were evaluated with the Comprehensive Assessment of Symptoms and History (CASH) and magnetic resonance imaging. Multiple correlation indicated that poor premorbid adjustment was significantly associated with prominence of negative symptoms, early age of onset, educational problems, chronicity, and neurological soft signs, but not with any MRI measures. These results confirm poor premorbid adjustment as an important predictor of a malignant form of schizophrenia as evidenced by an earlier age of onset, poorer educational performance, prominent negative symptoms, presence of soft signs, and chronicity of course. PMID- 7488566 TI - Durable improvements in Wisconsin Card Sorting Test performance in schizophrenic patients. AB - We examined the potential reversibility of cognitive deficits of schizophrenic patients in performing the Wisconsin Card Sorting Test (WCST), a neuropsychological measure of frontal lobe performance. The sample comprised 34 DSM III-R schizophrenic patients, who were randomly assigned to one of three groups, viz. two experimental groups and one control group, and were tested on six consecutive occasions. The results show that schizophrenic patients, selected on their poor WCST performance, could dramatically improve their performances by an elaborate procedural test training. They were even able to sustain these improvements over a 2-week period. The addition of monetary reinforcement to the test training was less effective than training alone. This reinforcement had an inhibiting effect on the training. Our results suggest that frontal lobe deficits in schizophrenia can be reduced and that a central deficit in schizophrenic WCST performance is a stable dysfunction in concept formation. For now it remains to be seen that this deficit in concept formation is remediable. PMID- 7488567 TI - Investigating thought disorder in schizophrenia with the lexical decision task. AB - Prior work has found evidence of hyperpriming in schizophrenics, which has been related to a persistence of associational activation in thought disorder. We extended this work by administering a lateralized version of the lexical decision task to 10 thought-disordered schizophrenics (TD), 10 non-thought-disordered schizophrenics (NTD), and 11 control subjects to localize the patterns of priming. All subjects were right-handed males. Associated, unassociated, and neutral prime-target pairs were displayed across seven stimulus location combinations: prime and target both presented to the center, left visual field (LVF), or right visual field (RVF); prime presented to the center with target presented to the LVF or RVF; and prime and target presented contralaterally. TD schizophrenics exhibited priming effects comparable to those of the other subject groups, suggesting intact semantic memory structures and memory processes that may be unimpaired under the conditions studied. It is believed that the associative process mediating priming effects found in schizophrenia are later in the sequence than those examined in this study. PMID- 7488568 TI - Relations between social skills and ward behavior in chronic schizophrenia. AB - The relationship between social skills and ward behavior among chronic schizophrenia patients was investigated. Twenty-eight inpatients participated in an unstructured role play test and were rated by staff members on a number of indices of ward behavior (e.g., social interactions, inappropriate behavior). Overall, there was a relationship between social skill on the role play and social behavior on the ward. Speech clarity was the social skill most strongly related to ward behavior, even when controlling for symptomatology. However, several other social skill variables (e.g., gaze, affect) were not related to social adjustment. The results are discussed in terms of the validity of social skill assessments in patients with chronic schizophrenia, and their implications for psychosocial treatment of this population. PMID- 7488569 TI - Does prenatal exposure to influenza in mice induce pyramidal cell disarray in the dorsal hippocampus? AB - Epidemiological studies point to an association between prenatal exposure to influenza and later schizophrenia. Such studies are consistent with neuropathologic reports demonstrating cytoarchitectural abnormalities in the hippocampus and parahippocampal gyrus suggestive of second trimester developmental anomalies. The hypothesis that prenatal exposure to influenza in the second trimester may induce hippocampal pyramidal cell disarray in mice was investigated. Between days 9-16 of pregnancy, 35 Balb/c mice were intranasally inoculated with either a mouse-adapted or non mouse-adapted pool of Influenza A/Singapore/1/57 (H2N2), and 10 controls were inoculated with normal saline. Offspring were sacrificed on day 21 postpartum. Microscopic examination of the CA1-CA2 junctional areas in the offspring of mice exposed to influenza failed to demonstrate excess pyramidal cell disarray when compared with influenza-free, age matched controls. There was evidence that disarray was greater among those exposed on day 13 of pregnancy. Analyses of the data by sex and severity of maternal infection failed to reveal any significant effects. PMID- 7488571 TI - Aqueous contamination during small incision cataract surgery: a lesson in study design. PMID- 7488570 TI - Proliferative vitreoretinopathy. PMID- 7488572 TI - Onset and recurrence of proliferative vitreoretinopathy in various vitreoretinal disease. AB - AIMS: This study aimed to evaluate both the mean time intervals between retinal disease and the development of proliferative vitreoretinopathy (PVR) and the time intervals at which recurrent PVR develops following various kinds of vitreoretinal disease and surgery. METHODS: One hundred and thirty six consecutive cases of PVR that were seen and operated on between 1991 and 1994 were evaluated retrospectively. Intervals between onset of disease and PVR or recurrent PVR were noted. Conditions leading to PVR, surgical procedures, and the final anatomical and functional results were evaluated. RESULTS: Before PVR was noted for the first time, an average of 1.06 vitreoretinal surgical procedures were performed (range 0 to 3). A mean of 1.99 surgeries were performed to control PVR or to reach an end stage where further surgery would have been unreasonable (range 0 to 5). The average (median) time interval between the onset of the retinal disease and PVR was 2.0 months (range 0.5 to 45 months). The median time intervals between surgery and recurrence for the second, third, or fourth times was also 2.0 months (range 0.5 to 34 months). CONCLUSIONS: PVR starts earliest at 2 weeks after an event and subsequently quietens down within a maximum of 45 months after treatment (median 2 months). PVR may recur more than once but the time course of the disease is likely to be the same for each recurrence. PMID- 7488574 TI - Refractive and visual results and patient satisfaction after excimer laser photorefractive keratectomy for myopia. AB - A study of excimer laser photorefractive keratectomy was performed on 108 patients. Variation in individual refractive outcome was noted, particularly for the higher levels of correction. At 12 months, of those with up to 3 dioptres of myopia, 93% were within 1 dioptre of emmetropia, 100% achieved an acuity of 6/12 unaided, 69% were very pleased with their result, and 10% were very disappointed. Of those with between 3.1 and 6 dioptres of myopia, the respective figures were 75%, 73%, 65%, and 11%. For those with over 6 dioptres of myopia the respective figures were 46%, 58%, 38%, and 23%. Some patients were disappointed despite having achieved good unaided visual acuity. Forty seven per cent of patients lost at least one Snellen line of best corrected acuity. Glare was a problem for some of the time in over 60%. Perceived distortion of vision was a problem for all of the time in 11% to 31% of cases depending on the initial level of myopia. In some cases the refraction continued to change between 12 months and 20 months. PMID- 7488573 TI - Anterior chamber aspirate cultures in small incision cataract surgery. AB - BACKGROUND: Anterior chamber aspirates on completion of extracapsular cataract surgery contain significant numbers of organisms, particularly coagulase negative staphylococci, an important cause of endophthalmitis. METHODS: Culture rates were compared in 50 patients after phacoemulsification surgery, which allows the possible benefits of a small, self sealing wound and maintenance of positive intraocular pressure, with a similar number of extracapsular cases. RESULTS: A culture positive rate of 20% and 24% respectively was found, an insignificant difference. CONCLUSION: Small incision surgery has no proved advantage over extracapsular surgery in terms of reducing the intraoperative bacterial inoculum. The significance of this result in terms of causation of endophthalmitis is discussed. PMID- 7488576 TI - Fundus imaging in patients with cataract: role for a variable wavelength scanning laser ophthalmoscope. AB - AIMS: An investigation was carried out to compare the image quality of the ocular fundus obtained clinically, photographically, and with the scanning laser ophthalmoscope (SLO) at visible and infrared wavelengths in patients with significant cataract. METHODS: Nineteen patients admitted for routine cataract extraction were examined clinically by two independent observers to ascertain cataract type and clarity of fundus view with an indirect ophthalmoscope. Fundus photography and both confocal and direct (non-confocal) SLO imaging at 590 nm, 670 nm, and 830 nm were carried out after pupillary dilatation. Images obtained were graded independently using a recognised grading system. RESULTS: Quality of SLO images appeared to be superior to indirect ophthalmoscopy (p < 0.01) and fundus photography (p < 0.001) when graded subjectively. Quantitative analysis of contrast of retinal vessels demonstrated significantly higher contrast for the SLO compared with digitised fundus photographs at all wavelengths tested (p < 0.001), with highest contrast at 590 nm. Use of a confocal aperture significantly improved vessel contrast but may reduce overall image intensity. CONCLUSIONS: Scanning laser ophthalmoscopy may offer a method to observe and record fine fundus detail in patients who have marked cataract. PMID- 7488575 TI - Paediatric post-traumatic endophthalmitis. AB - AIMS: A retrospective analysis of children with post-traumatic endophthalmitis was performed to determine if microbiological differences exist between this disease in the paediatric population compared with this disease in adults. METHOD: Twelve cases of post-traumatic endophthalmitis in children were analysed to determine characteristics of this disease in youth. Patient ages varied from 18 months to 13 years; the mean age was 8 years. Gram positive organisms were isolated in eight eyes, Gram negative organisms from four eyes, fungus from one eye, and negative cultures in three eyes. The most common isolates were streptococcal species (56.6%) and staphylococcal species (22.2%). Vitrectomy was performed on eight (66.7%) eyes. RESULTS: Visual acuity of 20/200 or better was obtained in eight eyes (66.7%). Three eyes had vision less than 5/200. One eye developed phthisis bulbi. Nine (75%) patients were younger than 10 years of age, and six (66.7%) of these nine obtained a final visual acuity of 20/200 or better. CONCLUSION: Useful vision can be obtained in children with post-traumatic endophthalmitis with early, aggressive treatment. The microbiology of paediatric post-traumatic endophthalmitis differs from adult disease with streptococcal species as the most common infecting organisms. PMID- 7488577 TI - Confocal fundus imaging with a scanning laser ophthalmoscope in eyes with cataract. AB - AIMS/BACKGROUND: The study aimed to determine the influence of increased intraocular light scatter on the contrast in scanning laser ophthalmoscope (SLO) images and to examine to what extent SLO images can visualise the fundus through media opacities due to cataract. METHODS: Intraocular light scatter was estimated from measurements of letter contrast sensitivity before and after cataract surgery in five eyes. SLO images were obtained before and after surgery using confocal apertures of 1, 2, 4, and 10 mm, at laser wavelengths of 633 and 780 nm. Visibility of the fundus was determined by measurements of retinal contrast. SLO images were compared with standard fundus photographs. RESULTS: SLO images obtained before surgery revealed details of the retina that were unresolvable in the fundus photographs because of light scattering. By using one of the three smallest apertures, image contrast was further improved. However, no simple relations between aperture size, estimated light scatter, and image contrast could be found. CONCLUSION: SLO imaging was found to be superior to fundus photography for viewing the retina in eyes with cataract. Owing to the inhomogeneous nature of cataracts, the optimal choice of confocal aperture and laser wavelength is not simple and must be individualised. PMID- 7488578 TI - Effects of argon (green) laser treatment of soft drusen in early age-related maculopathy: a 6 month prospective study. AB - AIM: To evaluate whether perifoveal laser photocoagulation of soft drusen reduces the total area occupied by drusen. METHOD: In a prospective, randomised study, 20 patients (mean age 71.3 (SD 7.7) years) with early age-related maculopathy (ARM) in the form of soft drusen and pigmentary changes and with good visual acuity (0.94 (0.09)) were treated with argon green laser photocoagulation. Mild laser burns (200 microns) were placed within a temporal horseshoe-shaped area, extending from a distance of no less than 500 microns from the centre of the fovea to the vascular arcades. They were placed on the drusen and scattered over areas where no drusen were present. Fundus colour photography and fluorescein angiography were performed at study entry as well as after 3 and 6 months. Nineteen age-matched patients (68.5 (6.2) years) with soft drusen maculopathy (visual acuity 0.95 (0.10)) were followed as a control group. Using a computer system, the area occupied by drusen was determined as a portion of the total area of a circle with a radius corresponding to 1250 microns in the fundus for the angiograms and 2500 microns for the colour photographs, respectively. At study entry, the mean area occupied by drusen in the treatment group was not significantly (p > 0.5-0.7) different from that of the control group. RESULTS: The mean drusen area of the angiograms and fundus colour photographs in the treated group decreased significantly from 19.3% to 13.8% (p = 0.002) and from 7.84% to 5.02% (p = 0.005), respectively. In the untreated group, the mean area occupied by drusen increased significantly from 17.4% to 20.4% (p = 0.030) and from 8.33% to 9.23% (p = 0.002), respectively. No significant changes in visual acuity (p > 0.05-0.5) or in visual fields (p > 0.05-0.3) occurred in either of the two groups during 6 months. CONCLUSION: The study suggests that the area occupied by soft drusen in patients with early ARM decreases after perifoveal laser photocoagulation but increases in an untreated control group. PMID- 7488579 TI - Optic disc measurement: a comparison of indirect ophthalmoscopic methods. AB - AIMS: Two methods of indirect ophthalmoscopic estimation of optic disc size, the 78 dioptre lens and optic disc biometer were evaluated. METHODS: Twenty nine eyes of 29 patients were measured by both methods and compared with optic disc size calculated using the three planimetric corrections described by Bengtsson and Krakau. RESULTS: The closest agreement with the clinical measurements was found using correction 3. There was a significant difference between both the 78 D lens (p < 0.0001) and the biometer (p = 0.0027) and the planimetric results. There was also a significant difference between the two clinical methods (p < 0.0001). Both methods showed acceptable intraobserver variation (CoV 2.45% and 3.13% respectively). CONCLUSION: Overall, both methods give larger measurements than planimetry; the 78 D lens by 0.41 mm and the biometer by 0.15 mm. Neither method gives a satisfactory estimation of optic disc size when compared with planimetry. PMID- 7488580 TI - Autosomal dominant cone-rod dystrophy with negative electroretinogram. AB - AIMS: The negative electroretinogram (ERG) is observed in many hereditary retinal disorders. However, no reports have described a negative ERG in a family with autosomal dominant cone-rod dystrophy. A Japanese family with autosomal dominant cone-rod dystrophy with negative ERG is described. METHOD: Members of a Japanese family with autosomal dominant cone-rod dystrophy were examined and evaluated with Goldmann and Humphrey perimetry, bright flash ERG with an intense white stimulus, rod, cone, and flicker ERGs, and fluorescein angiography. Molecular analysis of the rhodopsin and peripherin/RDS genes in the patients was also performed. RESULTS: A 45-year-old Japanese man (proband) presented with decreased visual acuity. His fundi revealed bull's eye maculopathy and his single flash bright ERG showed a negative configuration. Negative ERG responses also were found in his father, who had macular degeneration, and one of the proband's three children who showed no fundus changes. No irregularities were found in their rhodopsin or peripherin/RDS genes. CONCLUSION: The condition of this family is believed to represent a previously undescribed autosomal dominant cone-rod dystrophy. PMID- 7488581 TI - Diffuse retinal pigment epitheliopathy complicating systemic corticosteroid treatment. AB - AIMS/BACKGROUND: This study was undertaken to confirm the association between diffuse retinal pigment epitheliopathy (DRPE) and systemic corticosteroid therapy. This finding can be of help in determining an aetiological factor in DRPE and associated diseases. Corticosteroids may contribute to the development of leakage in the presence of a retinal pigment epitheliopathy or central serous chorioretinopathy. METHODS: Cases of DRPE were collected from the files of members of the European Fluorescein Angiography Club. There were 34 who developed their ophthalmic symptoms while being treated with corticosteroids. RESULTS: DRPE developed in 34 patients from 10 European eye clinics during systemic treatment with corticosteroids. Thirteen patients were treated with corticosteroids after allotransplantation, 21 other patients developed ocular symptoms while treated with steroids for serious systemic disorders. Symptoms occurred in some patients when the daily corticosteroid dosage was elevated, and the visual complaints ameliorated or even disappeared sometimes on discontinuation of the corticosteroid therapy. CONCLUSION: Corticosteroids can damage the retinal pigment epithelial barrier and predispose a patient to serous retinal detachment, whereas psychological stress may play a role in the development of central serous chorioretinopathy. A decrease of the daily corticosteroid dosage may help to diminish the visual symptoms. PMID- 7488582 TI - Ophthalmology in Luanda (Angola): a hospital based report. AB - AIMS/BACKGROUND: As part of a 4 year Spanish development aid programme, an ophthalmic hospital was set up in Luanda in 1991 for the in situ training of local ophthalmologists. Presented here are the data obtained from 4201 patients treated during the first 2 years of the project. METHODS: Patients were referred to the institute from the emergency ward at the Luanda General Hospital, selected mainly according to the severity of their disease. The following data were collected from the clinical reports: age, sex, diagnosis (single or multiple), type of treatment (medical or surgical), acuity of the best eye at the time of diagnosis, and main disease group. RESULTS: The main causes of blindness treated were: cataracts; glaucoma; optic nerve diseases (neuritis and atrophy); trauma; xerophthalmia; uveitis; hereditary retinal diseases (degenerative myopia, retinitis pigmentosa, albinism, and Stargardt's disease); retinal detachment; and diabetic retinopathy. CONCLUSIONS: Sanitary resources in Angola are generally inadequate, and ophthalmic care is no exception to this. Owing to the high percentage of preventable or treatable blinding diseases in this environment, a campaign of social education should always be held along with any medical programme, in order to optimise the available resources. PMID- 7488583 TI - Effect of the cytokines on the prostaglandin E2 synthesis by lens epithelial cells of human cataracts. AB - BACKGROUND: Lens epithelial cells (LECs) derived from human cataracts have been reported to produce various cytokines and prostaglandin E2 (PGE2) in culture. The effects of IL-1, TGF-beta, and b-FGF on the PGE2 synthesis by LECs have been studied. METHODS: A circular piece of the anterior capsule with attached LECs was obtained by capsulotomy during cataract surgery and cultured. The primary, almost confluent, cultures were used for the study. The PGE2 concentration of the culture media for 24 h was measured after the addition of recombinant human IL-1 alpha, TGF-beta 2, or b-FGF at various concentrations. The PGE2 concentration was also measured in the media to which each cytokine and rabbit polyclonal anti human antibodies against the corresponding cytokine had been added. RESULTS: The PGE2 concentration of the culture media after addition of IL-1 alpha at the concentration of 100 or 500 pg/ml (1765 (768) and 3071 (1121) pg/10(4) cells) or TGF-beta 2 at the concentration of 10 or 100 ng/ml (689 (264) and 750 (189) pg/10(4) cells) was significantly increased compared with that in the controls (67 (20) pg/10(4) cells). These effects were suppressed by the corresponding anticytokine antibodies. Basic FGF and anti-human b-FGF showed no significant effect on the PGE2 concentration. IL-1 and TGF-beta increased but b-FGF did not affect the PGE2 synthesis by LECs in culture. CONCLUSION: IL-1 and TGF-beta may participate in postoperative inflammation after cataract surgery by increasing PGE2 synthesis by residual LECs. PMID- 7488584 TI - Types of collagen synthesised by the lens epithelial cells of human cataracts. AB - AIMS/BACKGROUND: Residual lens epithelial cells (LECs) undergo fibrous proliferation after cataract surgery, resulting in capsular fibrosis. The purpose of this study was to determine the types of collagen produced in cultured LECs derived from human cataract LECs. METHODS: A circular section of the anterior capsule, about 5 mm in diameter, with LECs attached was obtained by anterior capsulotomy during cataract surgery and cultured directly without dispersion of the cells in a well, on the bottom of which a disc-shaped, thin plate of poly(methyl methacrylate) had been placed. At 5 to 6 weeks of culture, the proliferated cells of the culture were stained immunohistochemically with antibodies against human collagens I-VI by the avidin-biotin-peroxidase complex method. RESULTS: Collagens I, IV, V, and VI were positive in the cultured cells. Types IV and V were strongly present in almost all the cells whereas types I and VI were only observed in a few cells. Collagens II and III were negative. CONCLUSIONS: Since the lens capsule is known to be comprised of collagen IV, collagens I, V, and VI seem to be produced newly in culture. The capsular fibrosis seen after cataract surgery in vivo as a wound healing process of the lens capsule, may contain these types of collagens. The present culture model is useful for studying secondary cataract formation. PMID- 7488585 TI - Preparation of Bruch's membrane and analysis of the age-related changes in the structural collagens. AB - AIMS/BACKGROUND: The morphological changes in Bruch's membrane and its constituent collagen seen during aging have been studied extensively but the chemical nature of the collagen and any aging changes have not previously been evaluated. METHODS: A method for preparing purified Bruch's membrane from human cadaver eyes by dissection preceded by trypsin digestion was developed. Following pepsin digestion, the constituent collagens were analysed by SDS-PAGE and by immunoblotting. Cyanogen bromide digestion was used to ascertain the solubility of the collagen and the proportion of type I to type III collagen. After hydrolysis of Bruch's membrane samples the constituent amino acids and collagen crosslinks were measured. RESULTS: The presence of collagen types I, III, IV, and V in Bruch's membrane was confirmed. The proportion of type III collagen as a percentage of total fibrous collagens was calculated as being between 35% and 39%, with no significant difference between different macular and peripheral sites or with age. There was a highly significant decline in the solubility of Bruch's membrane collagen with age, from near 100% in the first decade of life to 40-50% in the ninth decade at both macular and peripheral sites. There was no significant change in the amount of enzymatically formed collagen crosslinks with age. Amino acid analysis indicated a significant increase in the amount of non collagen protein with age in macular but not peripheral sites. CONCLUSION: Changes in the constituent collagens may contribute to the accumulation of debris in Bruch's membrane with age and interfere with the function of the retinal pigment epithelium, with subsequent consequences for the overlying photoreceptors. PMID- 7488587 TI - Ultrasound biomicroscopic imaging of a surgically reattached cyclodialysis cleft. PMID- 7488589 TI - Ophthalmic services for children. PMID- 7488586 TI - Proliferative vitreoretinopathy: pathobiology, surgical management, and adjunctive treatment. PMID- 7488588 TI - Cytomegalovirus retinitis in an AIDS patient without severe depletion in CD4 cell count. PMID- 7488590 TI - Proliferation of lens epithelial remnants after Nd-YAG laser capsulotomy. PMID- 7488591 TI - Laser photodisruption of visible retinal artery emboli. PMID- 7488592 TI - Information fidelity in corneal topography. PMID- 7488593 TI - Modulation of amblyopia therapy following early surgery for unilateral congenital cataracts. PMID- 7488594 TI - Beta blockers and the elderly with glaucoma: are we adding insult to injury? PMID- 7488595 TI - EyeSys corneal topography measurement applied to calibrated ellipsoidal convex surfaces. AB - AIMS/BACKGROUND: This study was carried out to assess the accuracy of the EyeSys videokeratoscope by using convex ellipsoidal surfaces of known form. METHODS: PMMA convex ellipsoidal buttons were calibrated using Form Talysurf analysis which allowed subsequent calculation of the vertex radius and p value of the surface. The EyeSys videokeratoscope was used to examine the same ellipsoids. The tabular data provided by the instrument software were used to plot a graph of r2 versus y2 where r is the measured radius at y, the distance from the corneal point being measured to the surface vertex. The intercept on the ordinate of this graph gives the vertex radius and the slope the p value. The results arising from the Talysurf and the EyeSys techniques were compared. RESULTS: The EyeSys videokeratoscope gave readings for both vertex radius and p value that were higher than those of the Talysurf analysis. The vertex radius was around 0.1 mm greater. The p value results were similar by the two methods for p values around unity but the EyeSys results were higher and the discrepancy increased as the p value approached that of a paraboloid. CONCLUSIONS: Although the videokeratoscope may be useful in comparative studies of the cornea, there must be some doubt about the absolute values displayed. The disagreement is sufficiently large to suggest that the instrument may not be accurate enough for contact lens fitting purposes. PMID- 7488597 TI - Treatment for 'A' or 'V' pattern esotropia by slanting muscle insertion. AB - BACKGROUND: Patients with 'A' or 'V' pattern esotropia without vertical muscle overaction have traditionally been treated with medial rectus recession and vertical transposition of the muscle insertions. METHOD: Seven cases are presented treated by slanting muscle insertions, whereby the lower margin of the medial rectus is preferentially recessed more than the upper margin in esotropia, and the upper margin is recessed more than the lower margin in A esotropia. RESULTS: All seven patients had their A or V patterns eliminated, with six achieving good alignment in all positions of gaze. CONCLUSION: The slanting muscle insertion should be considered as an alternative technique in all cases of A or V pattern esotropia. PMID- 7488596 TI - Modulation of amblyopia therapy following early surgery for unilateral congenital cataracts. AB - BACKGROUND: Stimulus deprivation amblyopia is the principal cause of visual impairment in infants with unilateral congenital cataract. Even if lensectomy is undertaken at an early age, intensive postoperative occlusion of the phakic eye is essential for the development of useful vision in the aphakic eye. Despite this, the optimum method of regulating occlusion therapy is uncertain. METHODS: Interocular acuity differences identified using clinical preferential looking techniques (Keeler cards) were used to regulate target levels of phakic eye occlusion in a prospective evaluation of 10 systemically, metabolically, and neurologically normal infants in whom dense unilateral cataract was diagnosed before 8 weeks of age, and operated upon by 10 weeks. Actual occlusion levels were recorded each day by parents in a diary. The development of preferential looking acuity in the phakic and aphakic eye were compared with prediction intervals derived from observations on 43 normal children. RESULTS: Aphakic eye preferential looking acuities were within the normal range at last review in all but one infant. Interocular acuity differences were < or = 0.5 octave in all children older than 1 year of age at last review, and > or = 1 octave in three of four children less than 1 year old at last review (Fisher exact p = 0.033). Phakic eye acuities were within the normal range in all infants at all visits. CONCLUSION: Within the first 2 years of life, normal preferential looking acuity may be achieved in both eyes of infants undergoing early surgery for unilateral congenital cataract if occlusion therapy is modulated according to interocular acuity differences quantified by clinical preferential looking techniques. PMID- 7488599 TI - Low vision in east African blind school students: need for optical low vision services. AB - AIMS: There is increasing awareness of the needs of children with low vision, particularly in developing countries where programmes of integrated education are being developed. However, appropriate low vision services are usually not available or affordable. The aims of this study were, firstly, to assess the need for spectacles and optical low vision devices in students with low vision in schools for the blind in Kenya and Uganda; secondly, to evaluate inexpensive locally produced low vision devices; and, finally, to evaluate simple methods of identifying those low vision students who could read N5 to N8 print after low vision assessment. METHODS: A total of 230 students were examined (51 school and 16 university students in Uganda and 163 students in Kenya, aged 5-22 years), 147 of whom had a visual acuity of less than 6/18 to perception of light in the better eye at presentation. After refraction seven of the 147 achieved 6/18 or better. Eighty two (58.6%) of the 140 students with low vision (corrected visual acuity in the better eye of less than 6/18 to light perception) had refractive errors of more than 2 dioptres in the better eye, and 38 (27.1%) had more than 2 dioptres of astigmatism. RESULTS: Forty six per cent of students with low vision (n = 64) could read N5-N8 print unaided or with spectacles, as could a further 33% (n = 46) with low vision devices. Low vision devices were indicated in a total of 50 students (35.7%). The locally manufactured devices could meet two thirds of the need. CONCLUSION: A corrected distance acuity of 1/60 or better had a sensitivity of 99.1% and a specificity of 56.7% in predicting the ability to discern N8 print or better. The ability to perform at least two of the three simple tests of functional vision had a sensitivity of 95.5% and a specificity of 63.3% in identifying the students able to discern N8 or better. PMID- 7488600 TI - Macular grid laser photocoagulation in uveitis. AB - AIMS/BACKGROUND: The aim of this study was to evaluate whether grid laser photocoagulation of the macula is beneficial in the treatment of cystoid macular oedema in patients with uveitis. METHODS: Six eyes of five patients with long standing cystoid macular oedema due to chronic uveitis were treated by grid laser photocoagulation of the macula. RESULTS: In the first weeks after treatment a temporary increase of oedema and paracentral scotomas were observed. At the long term follow up of more than 18 months in all patients, macular oedema had been reduced significantly or disappeared in all eyes treated. One eye had a significant increase in Snellen acuity, three eyes more or less stabilised, and two eyes deteriorated. CONCLUSION: The beneficial effect of laser treatment on visual acuity in patients with uveitis might be more favourable if performed at an earlier stage of the disease. PMID- 7488598 TI - Intraocular pressure rise after phacoemulsification with posterior chamber lens implantation: effect of prophylactic medication, wound closure, and surgeon's experience. AB - AIMS: A prospective clinical trial was carried out to evaluate the effect of prophylactic medication, the technique of wound closure, and the surgeon's experience on the intraocular pressure rise after cataract extraction. METHODS: In 100 eyes, the intraocular pressure was measured before as well as 2-4, 5-7, and 22-24 hours after phacoemulsification and posterior chamber lens implantation. Each of 25 patients received either 1% topical apraclonidine, 0.5% topical levobunolol, 500 mg oral acetazolamide, or placebo. Forty four eyes were operated with sclerocorneal sutureless tunnel and 56 eyes with corneoscleral incision and suture. Sixty three operations were performed by experienced surgeons (more than 300 intraocular operations) and 37 by inexperienced surgeons (less than 200 intraocular operations). RESULTS: The pressure increase from baseline to the maximum 5-7 hours after surgery did not differ significantly (p = 0.8499) for apraclonidine (9.5 mm Hg), levobunolol (7.2 mm Hg), acetazolamide (7.8 mm Hg), and placebo (8.6 mm Hg). The increase was significantly (p = 0.0095) lower in eyes with corneoscleral tunnel (5.5 mm Hg) than in eyes with corneoscleral suture (10.5 mm Hg) and significantly (p = 0.0156) lower for experienced (6.6 mm Hg) than for inexperienced surgeons (11.2 mm Hg). CONCLUSIONS: The intraocular pressure rise after phacoemulsification and posterior chamber lens implantation depends strongly on the technique of wound closure and the surgeon's experience. Compared with these two factors, the effect of prophylactic medication can be neglected. PMID- 7488601 TI - New Dacron tissue colonisable keratoprosthesis: clinical experience. AB - BACKGROUND: Keratoprostheses (KPs) are made of an optical cylinder integrated with a supporting element which conditions their biocompatibility. A new KP with a Dacron tissue colonisable support has been designed in order to reduce significantly the complication rate. METHODS: This new KP was implanted into 20 eyes of 20 patients with bilateral corneal blindness unsuited to a corneal implant. The follow up ranged from 24 to 96 months. RESULTS: All 20 patients had some improved visual acuity with 13 retaining this improvement for more than 2 years. CONCLUSION: These favourable results may be indicative of the reliability of the new KP. PMID- 7488603 TI - Flow cytometric identification of a minority population of MHC class II positive cells in the normal rat retina distinct from CD45lowCD11b/c+CD4low parenchymal microglia. AB - AIMS: This study aimed to isolate and classify by flow cytometry, the cell surface phenotype of microglia in the normal rat retina with a view to identifying putative antigen presenting cells (APC) within the retina, which has to date not been possible by immunohistochemistry. METHODS: Normal rat retinal microglia were isolated and classified using a modification of an isolation technique employing graduated Percoll density gradient cell separation and flow cytometric phenotypic criteria used for CNS microglia. RESULTS: Retinal microglia can be defined by flow cytometry on the basis of their CD45lowCD11b/c+CD4low cell surface expression. Constitutive MHC class II expression in the normal rat retina was confined almost exclusively to a very minor population of cells expressing neither low (microglia) nor high levels of CD45. Three colour flow cytometric analysis confirmed that these MHC class II positive cells were ED2+. CONCLUSIONS: Using this sensitive isolation technique we have identified the cell surface characteristics of ramified, resident microglia, and found that they do not constitutively express MHC class II. There is, however, constitutive MHC class II expression on a phenotypically distinct population of cells (CD45low/highED2+). We propose these cells are the counterpart of the perivascular macrophages found in the CNS which present antigen to extravasating T cells, although their exact retinal location can only be confirmed by immunohistochemical analysis. The role of parenchymal microglia as APC remains undefined. Future isolation of microglia and putative perivascular cells using this technique will help identify the role these cells play in the initiation and perpetuation of immune responses within the retina. PMID- 7488604 TI - Bimodal expressivity in dominant retinitis pigmentosa genetically linked to chromosome 19q. AB - A clinical, psychophysical, and electrophysiologic study was undertaken of two autosomal dominant retinitis pigmentosa pedigrees with a genetic mutation assigned to chromosome 19q by linkage analysis. Members with the abnormal haplotype were either symptomatic with adolescent onset nyctalopia, restricted visual fields, and non-detectable electroretinographic responses by 30 years of age, or asymptomatic with normal fundus appearance and minimal or no psychophysical or electroretinographic abnormalities. There was no correlation in the severity in parents and their offspring. Pedigree analysis suggested that although the offspring of parents with the genetic mutation were at 50% risk of having the genetic defect, the risk of being symptomatic during a working lifetime was only 31%. Such bimodal phenotypic expressivity in these particular pedigrees may be explained by a second, allelic genetic influence and may be a phenomenon unique to this genetic locus. Genetic counselling in families expressing this phenotype can only be based on haplotype analysis since clinical investigations, even in the most elderly, would not preclude the presence of the mutant gene. PMID- 7488602 TI - Use of Proplast II as a subperiosteal implant for the correction of anophthalmic enophthalmos. AB - BACKGROUND: A variety of autogenous and alloplastic materials have been used as subperiosteal implants to correct anophthalmic enophthalmos. Proplast II is a synthetic porous composite of Teflon polymer and alumina. Proplast II offers a number of advantages over other commonly used alloplastic materials such as silicone and polymethyl methacrylate. It is light, porous, resilient, malleable, and easy to shape. It can be readily sterilised after shaping. It has been found to integrate with the surrounding tissues, thereby minimising the risk of subsequent implant migration and extrusion. METHODS: Proplast II was used as a subperiosteal implant in a total of 15 anophthalmic patients during the period June 1990 to March 1994. The indication for this procedure in all patients was poor orbital volume replacement despite the prior insertion of an adequately sized spherical socket implant. RESULTS: The results were excellent with a good correction of preoperative upper eyelid sulcus deformity. There were no operative complications nor any serious postoperative complications. The implants were well tolerated. CONCLUSION: Proplast II can be highly recommended for use as a subperiosteal implant. PMID- 7488605 TI - Effects of interferon alfa and gamma on human uveal melanoma cells in vitro. AB - BACKGROUND: Uveal melanoma is a tumour with a high incidence of metastasis and a high mortality rate. Additional therapies to obtain a better local control or an effective treatment of metastases are necessary. Interferons may be applied. METHODS: The effects of human interferon alfa and gamma on proliferation and expression of immunologically important molecules of human uveal melanoma cells in vitro were studied. A propidium iodide assay was used to determine proliferation and immunostaining with monoclonal antibodies was applied to detect changes in antigen expression on two primary uveal melanoma cell lines, Mel 202 and 92-1. RESULTS: Interferon alfa inhibited proliferation of cell line 92-1 at a concentration of 50 IU/ml, but had no effect on cell line Mel 202, while interferon gamma inhibited growth of both cell lines. Only interferon gamma had a visible effect on cell morphology. With respect to the immunomodulatory effects, interferon alfa increased monomorphic HLA class I expression, but did not affect HLA class II expression. Interferon gamma induced not only HLA class I but also class II expression. The effects on HLA expression were locus-specific with the strongest effects observed for HLA-B and DR products. Small differences were observed with respect to the susceptibility of two different melanoma cell lines to antiproliferative effects and to modulation of antigen expression. CONCLUSION: The effects of interferon alfa and gamma on human uveal melanoma cells in vitro suggest a potential role of these cytokines in the treatment of patients with uveal melanoma. In particular, the immunomodulatory effects of these cytokines in vitro imply that treatment of patients with these cytokines might stimulate a beneficial antimelanoma immune response in vivo. PMID- 7488607 TI - Peripheral retinal neovascularisation associated with scleral encirclement. PMID- 7488606 TI - Fundal white dots: the spectrum of a similar pathological process. PMID- 7488608 TI - Subretinal Pseudomonas abscess after lung transplantation. PMID- 7488609 TI - Peter's anomaly in an infant with a mild degree of Axenfeld's anomaly. PMID- 7488610 TI - Subretinal hypopyon in prolymphocytic leukaemia. PMID- 7488611 TI - Pseudomonas keratitis associated with biofilm formation on a disposable soft contact lens. PMID- 7488612 TI - Bilateral vitreous cysts in an 80-year-old man. PMID- 7488613 TI - Bacterial endophthalmitis after anterior chamber paracentesis. PMID- 7488614 TI - Contrast sensitivity testing in clinical practice. PMID- 7488615 TI - Changes in physical properties of vacuolar membrane during transformation of protein bodies into vacuoles in germinating pumpkin seeds. AB - Changes in membrane molecular dynamics associated with the transformation of protein body membranes into vacuolar membranes during pumpkin seed germination, were monitored by EPR-spin probe technique. Using highly purified membrane preparations as well as 5-SASL and 16-SASL spin labels, parameters like general membrane lipid fluidity, order parameter, semicone angle, rotational correlation times tau 2B and tau 2C, ratio of immobilized to mobile lipids were determined and the activation energy for rotational diffusion of 16-SASL was calculated. Analysis of these parameters at different temperatures indicated a more rigid nature of protein body membrane comparing to vacuolar membrane, as a result of a more restricted motional freedom of lipids. These differences are discussed in terms of protein composition and various functional specialization of both types of membranes. PMID- 7488617 TI - Modelling packing interactions in parallel helix bundles: pentameric bundles of nicotinic receptor M2 helices. AB - The transbilayer pore of the nicotinic acetylcholine receptor (nAChR) is formed by a pentameric bundle of M2 helices. Models of pentameric bundles of M2 helices have been generated using simulated annealing via restrained molecular dynamics. The influence of: (a) the initial C alpha template; and (b) screening of sidechain electrostatic interactions on the geometry of the resultant M2 helix bundles is explored. Parallel M2 helices, in the absence of sidechain electrostatic interactions, pack in accordance with simple ridges-in-grooves considerations. This results in a helix crossing angle of ca. +12 degrees, corresponding to a left-handed coiled coil structure for the bundle as a whole. Tilting of M2 helices away from the central pore axis at their C-termini and/or inclusion of sidechain electrostatic interactions may perturb such ridges-in grooves packing. In the most extreme cases right-handed coiled coils are formed. An interplay between inter-helix H-bonding and helix bundle geometry is revealed. The effects of changes in electrostatic screening on the dimensions of the pore mouth are described and the significance of these changes in the context of models for the nAChR pore domain is discussed. PMID- 7488616 TI - Changes in the passive electrical properties of human stratum corneum due to electroporation. AB - The stratum corneum (SC) is the main barrier to molecular and ionic transport across mammalian skin and has been extensively studied by others at low voltages (U(skin)(t) < 10 V) in order to partially characterize the skin. Here we use one or more exponential pulses (tau pulse = 1 ms) and a temperature of 25 +/- 2 degrees C and found that the low voltage passive electrical properties (impedance) change rapidly and significantly if these pulse result in U(skin),0 > 40 V. In contrast, the dynamic resistance (describing passive electrical behavior in a nonlinear range) changes dramatically by application of pulses between 40 V and 80 V and then it settles at levels between 50 omega and 100 omega. We also found that recovery of the low voltage electrical parameters after pulsing depends mainly on the voltage, and, for multiple pulse protocols, on the number of pulses. For single pulses of U(skin),0 approximately 90 V or less the electrical recovery was almost complete, returning to within 0.90 of the pre pulse value. In contrast, larger pulses result progressively in decreased recovery. The recovery for pulses > 90 V revealed several characteristic times, suggesting the involvement of different processes. For multiple pulses with U(skin),0 > 130 V almost no recovery of the transdermal resistance, R(skin), was evident (returning to < 0.10 of pre-pulse values), i.e., essentially permanent changes in the stratum corneum occurred. This is similar to that of single bilayer membrane electroporation, for which a transition from reversible to irreversible behavior occurs as transmembrane voltage is increased. Thus, these results are consistent with the hypothesis that 'high-voltage' pulses cause electroporation within the SC, i.e., that elevated transmembrane voltage result in creation of new aqueous pathways ('pores') across SC lipid regions. PMID- 7488618 TI - Attachment of antibodies to sterically stabilized liposomes: evaluation, comparison and optimization of coupling procedures. AB - Several coupling methods for binding antibodies (Ab) to liposomes have previously been developed. We were interested in examining if some of these methods would be suitable for attaching Ab to long-circulating formulations of liposomes (SL), sterically stabilized with poly(ethylene glycol) (PEG). We studied three 'classical' coupling methods in which Ab was attached at the bilayer surface of SL, and two new coupling methods in which Ab was attached at the PEG terminus. Parameters examined including binding efficiency, antibody surface density, the ability of the immunoliposomes to remote-load the anticancer drug doxorubicin, and the specific binding of the resulting immunoliposomes to target cells. The non-covalent biotin-avidin coupling method resulted in low Ab densities at the cell surface, as did a coupling in method in which maleimide-derivatized Ab was attached to the liposome surface through a thiolated phospholipid incorporated into the liposomes. The low levels of Ab achieved in these method was likely due to interference by PEG with the access of the Ab to the liposome surface. However, when a maleimide-derivatized Ab was coupled to thiolated PEG, moving the coupling reaction away from the liposome surface, very high coupling efficiencies were achieved, and these immunoliposomes achieved good specific binding to their target cells. Oxidizing the Fc region of the Ab and coupling it to the PEG terminus through a hydrazone bond was a less efficient coupling method, but had the advantage of retaining Ab orientation. Efficient remote-loading of doxorubicin was found for immunoliposomes in which Ab was attached at the PEG terminus. PMID- 7488619 TI - Transmembrane gradient driven phase transitions within vesicles: lessons for drug delivery. AB - Phase transitions in closed vesicles, i.e., microenvironments defined by the size of the vesicle, its contents, and permeability of its membrane are becoming increasingly important in several scientific disciplines including catalysis, growth of small crystals, cell function studies, and drug delivery. The membrane composed from lipid bilayer is in general impermeable to ions and larger hydrophilic ions. Ion transport can be regulated by ionophores while permeation of neutral and weakly hydrophobic molecules can be controlled by concentration gradients. Some weak acids or bases, however, can be transported through the membrane due to various gradients, such as electrical, ionic (pH) or specific salt (chemical potential) gradients. Upon permeation of appropriate species and reaction with the encapsulated species precipitation may occur in the vesicle interior. Alternatively, these molecules can also associate with the leaflets of the bilayer according to the transmembrane potential. Efficient liposomal therapeutics require high drug to lipid ratios and drug molecules should have, especially when associated with long circulating liposomes, low leakage rates. In this article we present very efficient encapsulation of two drugs via their intraliposomal precipitation, characterize the state of encapsulated drug within the liposome and try to fit the experimental data with a recently developed theoretical model. Nice agreement between a model which is based on chemical potential equilibration of membrane permeable species with experimental data was observed. The high loading efficiencies, however are only necessary but not sufficient condition for effective therapies. If adequate drug retention within liposomes, especially in the case of long-circulating ones, is not achieved, the therapeutic index decreases substantially. Anticancer drug doxorubicin precipitates in the liposome interior in a form of gel with low solubility product and practically does not leak out in blood circulation in the scale of days. With an antibiotic, ciprofloxacin, the high loading efficacy and test tube stability is not reproduced in in vitro plasma leakage assays and in vivo. We believe that the reasons are higher solubility product of precipitated drug in the liposome, larger fraction of neutral molecules due closer pK values of the drug with the pH conditions in the solutions and high membrane permeability of this molecule. High resolution cryoEM shows that encapsulated anticancer agent doxorubicin is precipitated in the form of bundles of parallel fibers while antibiotic ciprofloxacin shows globular precipitate. Doxorubicin gelatin also causes the change of vesicle shape. PMID- 7488620 TI - A novel strategy affords high-yield coupling of antibody Fab' fragments to liposomes. AB - A new assay for the production of reactive sulfhydryl-bearing antibody Fab' fragments has been utilized to develop conditions affording high efficiencies of coupling of mouse and rabbit IgG-derived Fab' fragments to lipid vesicles containing maleimidyl-functionalized phospholipids. Cysteine and mercaptoethylamine, but not dithiothreitol, reduce antibody F(ab')2 to Fab' fragments in very good yields under conditions where overreduction to heavy and light chains is minimized. Surprisingly, however, a large fraction of the Fab' fragments generated under these conditions can lack maleimide-reactive sulfhydryl groups, as demonstrated using a maleimidyl-poly(ethylene glycol) conjugate to shift selectively the electrophoretic mobility of the reactive sulfhydryl-bearing Fab' fragments. After modification of F(ab')2 reduction conditions specifically to maximize the yield of the latter fraction, it is possible to achieve high and very reproducible coupling of functional Fab' fragments to liposomes (equivalent to coupling of ca. 70% of total input protein and almost 100% of the reactive sulfhydryl-bearing Fab' fraction). A novel phospholipid-poly(ethylene glycol) maleimide 'anchor' allows particularly efficient coupling of Fab' fragments to liposomes, even using relatively low liposome concentrations and molar percentages of the liposome-incorporated 'anchor' species. These results demonstrate that with appropriate optimization of the conditions for Fab' production and liposome coupling, Fab' fragments can be coupled to liposomes with efficiencies comparable to or exceeding those reported for coupling of intact antibodies. These results should facilitate the wider use of Fab' fragments as a potentially advantageous alternative to intact antibodies for liposomal targeting in various applications. PMID- 7488621 TI - Rapid diffusion of the lipid phosphorus of phosphatidylglycerol liposomes through polycarbonate membranes is caused by the oxidation of the unsaturated fatty acids. AB - The lipid phosphorus of phosphatidylglycerol liposomes was found to diffuse extensively, after a lag time of 1 to 2 days, through a 0.1 micron pore size polycarbonate membrane in a two compartment system. Diffusion occurred when either multilamellar or large unilamellar vesicles were studied, even if they were sedimented to eliminate any smaller particles. The lipid of liposomes prepared under sterile conditions also diffused extensively. Diffusion appeared to be related to the age of the vesicles, and could be eliminated by incorporating antioxidants into the liposomes, or by using liposomes prepared from saturated phospholipids (C14 or larger). This indicated that diffusion was caused by phospholipid oxidation, which was confirmed by HPLC analysis. Phospholipid phosphorus that diffused through a membrane appeared more polar, as indicated by its capacity to distribute into the upper phase of a two phase extraction. Phospholipid phosphorus diffusion was preceded by the complete loss of liposomes contents, indicated by the complete diffusion of encapsulated carboxyfluorescein through the membrane. Oxidation of the lipid could be prevented by inclusion of either butylated hydroxytoluene or alpha-tocopherol in the membrane. The best retention of liposomal contents was achieved when both antioxidants and cholesterol were included in the liposome preparation. The antioxidant incorporated in the liposomes remained effective in protecting the phospholipids upon storage at 4 degrees C for 2 months. The inclusion of EDTA in the suspension medium retarded the rapid oxidation, suggesting that the presence of trace amounts of heavy metal ions in the buffer catalyzed the oxidation. Phospholipid oxidation was most effectively inhibited by the presence of serum or chemically defined medium, suggesting that oxidation of liposomal lipids in a biological environment may be minimized if appropriate steps are taken. PMID- 7488622 TI - An early transient increase of intracellular Na+ may be one of the first components of the mitogenic signal. Direct detection by 23Na-NMR spectroscopy in quiescent 3T3 mouse fibroblasts stimulated by growth factors. AB - 23Na-NMR spectroscopy was designed to allow for continuous recording of intracellular Na+ in 3T3 fibroblasts stimulated by serum growth-factors in the presence of ion transport inhibitors. The metabolic state of cells at rest and following stimulation was monitored by 31P-NMR spectra of ATP and related high energy phosphates. The study demonstrates that early activation of ion transporters by addition of serum is marked by the appearance of transient increase of the intracellular Na+, beginning 3 min after addition of serum to quiescent culture and lasting approx. 20 min. The initial rise in cellular Na+ results from an increased activity of the bumetanide-sensitive Na+/K+/Cl- cotransport and of the amiloride-sensitive Na+/H+ antiport. It is suppressed by any one of these inhibitors. Subsequent activation of the ouabain-sensitive Na+/K(+)-ATPase results in an increased Na+ efflux, leading to a return of intracellular Na+ to its initial baseline. Previous work had shown that the early activation of bumetanide-sensitive and amiloride sensitive ion-transporters by growth-factors was essential for induction of cell division, at least in some cell types. Preventing ion activation by adding ion-transport inhibitors lead to the inhibition of DNA synthesis 18 h later. This process was reversible upon elimination of these inhibitors. Even though alternative non-specific effects of these inhibitors cannot be ruled out, the observed transient peak in intracellular Na+ may be one of the earliest components of the mitogenic signal. On the basis of previous works, its effect seems to be related to the activation of Ca(2+)-dependent and cyclic AMP second messenger pathways. The different mechanisms whereby the activated Na+/K+/Cl- cotransport and the Na+/H+ antiport contribute to this signal need to be further investigated. PMID- 7488623 TI - Influence of monoglucosyldiacylglycerol and monoacylmonoglucosyldiacylglycerol on the lipid bilayer of the membrane from Acholeplasma laidlawii strain A-EF22. AB - The ability for 1,2-diacyl-3-O-(alpha-D-glucopyranosyl)-sn-glycerol (MGlcDAG) and 1,2-diacyl-3-O-(6-O-acyl-(alpha-D-glucopyranosyl))-sn-glycerol (MAMGlcDAG) to induce non-lamellar phases in a lipid mixture with an in vivo composition, prepared from Acholeplasma laidlawii membranes, has been investigated. The phase transition temperatures from lamellar to non-lamellar structures were studied with varying fractions of MGlcDAG and MAMGlcDAG. The transition temperature decreased from 73 +/- 2 degrees C for 20 mol% MGlcDAG to 43 +/- 1 degree C for 63 mol% MGlcDAG, in lipid mixtures where the other lipids are the native bilayer forming lipids. MAMGlcDAG behaved differently and the phase transition temperatures were found to be almost constant and between 51-53 degrees C as the fraction of MAMGlcDAG varied between 11-45 mol%. It was also found that MAMGlcDAG can only be solubilized in low concentrations in the lipid bilayer, which is in good agreement with the fractions of MAMGlcDAG found in the membrane of A. laidlawii. Higher concentrations of MAMGlcDAG resulted in phase separations of lamellar liquid crystalline and gel/crystalline phases. It is concluded that MAMGlcDAG is far more capable than MGlcDAG to induce non-lamellar structures at lower concentrations. The results are discussed in terms of the model of lipid regulation previously proposed by this laboratory (Lindblom, G., Hauksson, J.B., Rilfors, L., Bergenstahl, B., Wieslander, A. and Eriksson, P.O. (1993) J. Biol. Chem. 268, 16198-16207), and the importance for the bilayer stability in cell membranes. It is proposed that the phase behaviour of the membrane lipids has far reaching consequences for membrane function. PMID- 7488624 TI - Membrane fluidity of Escherichia coli during heat-shock. AB - The excimer-forming fluorophore dipyrenylpropane has been used to measure the relative fluidity of total membranes isolated from Escherichia coli grown at 30 or 45 degrees C, or exposed to a heat-shock from 30 to 45 degrees C for various periods of time. Parallel experiments were performed using [35S]methionine pulse labeling of cells, to study the induction of heat-shock proteins (HSPs) at different times after the sudden change in E. coli growth-temperature from 30 to 45 degrees C. Results suggest that upon an abrupt temperature upshift from 30 to 45 degrees C, membrane fluidity adjustment to the steady-state level at the high temperature, takes place during the E. coli heat-shock response. PMID- 7488625 TI - Amiloride-sensitive Na+ conductance in native Xenopus oocytes. AB - Endogenous Na+ conductances in the plasma membrane of oocytes of the South African clawed toad Xenopus laevis were investigated by microelectrode techniques and influx measurements. Removal of Na+ from the bath solution under voltage clamp conditions led to a decrease in the clamp current indicating the existence of native Na+ conductances. The observed current was voltage dependent but showed no marked rectification. Amiloride (10 microM) blocked this Na+ current reversibly. However, amiloride analogues such as benzamil and phenamil had no effect on this Na+ conductance. The Na+/H(+)-exchanger blocker EIPA (5-(N-ethyl-N isopropyl)amiloride), another amiloride analogue, also had no effect thereby excluding a possible involvement of the Na+/H+ exchanger. The Na+ mediated current had a reversal potential of about 50 mV suggesting high selectivity of these Na+ conductances for Na+ over other monovalent cations. When Na+ was replaced by K+ in the bath solution, amiloride had no effect on the clamp current over the whole potential range demonstrating that only Na+ but not K+ can enter the cell via the investigated conductances. In radio tracer experiments 22Na+ influx into oocytes was nearly halved in presence of amiloride (10 microM), whereas benzamil and phenamil again failed to influence 22Na+ influx. These results suggest that the endogenous amiloride-sensitive Na+ conductance belongs to a new class of channels which is quite different from amiloride-sensitive epithelial Na+ channels. PMID- 7488626 TI - Injury of rat thymocytes caused by exogenous peroxyl radicals in vitro. AB - The aim of this study was to investigate oxidative cell injury in rat thymocytes under conditions of radical generation exterior to the cell utilizing the thermolabile azocompound 2,2'-azobis(2-amidinopropane) dihydrochloride to generate peroxyl radicals at a constant and reproducible rate. This initiator, being water-soluble and endowed with a positive charge, is suitable for studies on oxidative damage of biomembranes induced in the external water environment. The relationship between cell viability, lipid and thiol oxidation and chain breaking antioxidant depletion was studied. During the first hour of treatment cell viability decreased slightly, protein sulfhydryl groups were consumed slowly and no significant production of conjugated dienes occurred. After 90 min of incubation, when thymocyte permeability started to increase, the concentration of alpha-tocopherol decreased gradually, significant changes of polyunsaturated fatty acids occurred and a rapid phase of thio oxidation commenced. It can be concluded that, under conditions of an exogenous oxidant challenge, initially the cell membrane provides a physical barrier to the entrance of radicals to the thymocyte. When peroxyl radicals gain access to the membrane and the molecular barrier begins to disorganize, the oxidizable cellular components become susceptible to massive attack. PMID- 7488627 TI - Metastability of dimiristoylphosphatidylethanolamine as studied by FT-IR and the effect of alpha-tocopherol. AB - The metastability of dimiristoylphosphatidylethanolamine (DMPE) has been studied by means of Fourier transform infrared spectroscopy (FT-IR), both in the absence and in the presence of alpha-tocopherol. Two different methods of hydration were used to prepare the samples, poorly hydrated and well hydrated, and the results have been compared with anhydrous DMPE. Poorly hydrated DMPE gave place to a high melting phase formed upon melting from gel to L alpha at approx. 49 degrees C, with a new transition to L alpha at approx. 55 degrees C. However, well hydrated DMPE incubated at 4 degrees C for 49 days gave place to a subgel phase which was transformed by heating into a L beta phase at about 40 degrees C and this into a L alpha phase after further heating at 52 degrees C. The subgel phase was more hydrated and less rigid than the high-melting phase. On the other hand, alpha tocopherol, when included in poorly hydrated DMPE, stabilized a high-melting phase, which was transformed by heating, directly into a L alpha. However, when a sample of DMPE containing alpha-tocopherol was incubated for 49 days at 4 degrees C a dehydrated solid phase different from the subgel and the high-melting phases was formed. PMID- 7488628 TI - Effects of plant sterols on the hydration and phase behavior of DOPE/DOPC mixtures. AB - Freeze-induced injury of protoplasts of non-acclimated rye and oat is associated with the formation of the inverted hexagonal (HII) phase in regions where the plasma membrane and various endomembranes are brought into close apposition as a result of freeze-induced dehydration. The influence of lipid composition and hydration on the propensity of mixtures of DOPE:DOPC containing either sterols or acylated steryl glucosides to form the HII phase was determined by DSC, freeze fracture electron microscopy and X-ray diffraction. The addition of plant sterols to a mixture of DOPE/DOPC (either 1:1:1 or 1:1:2 mole ratio of DOPE/DOPC/sterols) reduced the total hydration of the mixture (expressed as wt% water) after desorption over a range of osmotic pressures of 2.8 to 286 MPa. However, most or all of the water remaining in the dehydrated lipid mixtures was associated predominantly with the phospholipids. Both sterols and acylated steryl glucosides significantly promoted both the dehydration-induced and thermally induced L alpha ->HII phase transitions in DOPE/DOPC mixtures however, acylated steryl glucosides were much more effective. In mixtures containing plant sterols, the HII phase occurred after dehydration at 20 MPa (20 degrees C), which resulted in a water content of 11.7 wt%. In contrast, mixtures containing acylated steryl glucosides were in the HII phase in excess water, i.e., they did not require dehydration to effect the L alpha-->HII phase transition. The results indicate that genotypic differences in the lipid composition of the plasma membrane of rye and oat leaves have a significant influence on the propensity for formation of the HII phase during freeze-induced dehydration. PMID- 7488629 TI - Determination of binding constants of Ca2+, Na+, and Cl- ions to liposomal membranes of dipalmitoylphosphatidylcholine at gel phase by particle electrophoresis. AB - The zeta potentials of 1,2-dipalmitoyl-sn-glycero-3-phosphatidylcholine (DPPC) liposomes were measured at a gel phase as a function of CaCl2 concentration (0 200 mM) in a solution containing different NaCl concentrations (0-200 mM). The data obtained were analyzed with the diffuse double layer theory including the Graham theory. The intrinsic binding constants of ions to DPPC membranes and the distance of the shear plane have been determined independent of both the concentration of CaCl2 and that of NaCl. The values of the constants were 37 M-1 for Ca2+, 0.28 M-1 for Cl, and 0.25 M-1 for Na+; the distance was 0.24 nm under the assumption of binding stoichiometry of Ca2+/DPPC = 1:1. PMID- 7488631 TI - Effects of the anesthetic steroid alphaxalone and its inactive delta 16-analog on the thermotropic properties of membrane bilayers. A model for membrane perturbation. AB - We have studied in detail the effects of the anesthetic steroid alphaxalone and its inactive analog delta 16-alphaxalone on the thermotropic properties of model membranes using differential scanning calorimetry (DSC). The results obtained showed that, for model membranes from hydrated dipalmitoylphosphatidylcholine (DPPC), dioleoylphosphatidylcholine (DOPC), and egg sphingomyelin, the biologically active analog significantly broadened the phase transition, in contrast to the inactive one which produced only marginal effects. Also, alphaxalone abolished the pretransition in these preparations whereas its delta 16-analog only broadened it. However, in DPPE bilayers almost no differences were observed in the effects produced by the two analogs. These results suggest that the ability of the two steroids to perturb membranes is lipid dependent. Comparisons between the effects of the two steroids on lipid/cholesterol model membranes revealed that delta 16-alphaxalone excluded cholesterol from lipid/cholesterol/delta 16-alphaxalone ternary systems whereas alphaxalone enhanced the effects of cholesterol and reduced the cooperativity in the binary phospholipid/cholesterol system. In an attempt to determine whether the different thermotropic effects of the two steroids on model membranes were due to (a) differences in their ability to perturb the bilayers; (b) different extents of incorporation into the bilayer, solid state 2H-NMR was applied using specifically deuterated steroids. The 2H-NMR data showed that alphaxalone incorporated fully into the membrane bilayer up to a molar concentration of 20%, while its inactive analog did only up to a concentration of 1%. To compare the abilities of the two steroids to perturb membrane preparations when both analogs were present in equal amounts in the membrane, the effects of very low steroid concentrations on DPPC bilayers were studied using DSC. The experiment showed that alphaxalone perturbed the membrane bilayers more effectively than its inactive analog. These results strongly suggest that the small structural differences between the two steroids are responsible for the observed differences in their abilities to perturb membranes, possibly because of differences in the packing of these two molecules within the bilayers. PMID- 7488630 TI - Membrane permeabilization by alpha-helical peptides: a flow cytometry study. AB - The permeabilization by alpha-helical peptides of nucleated mammalian cells can be monitored by flow cytometry. Ethidium bromide, a non fluorescent and poorly membrane permeant molecule, becomes strongly fluorescent only upon binding to DNA. On this basis, the permeabilization of the plasma membrane of HL60 promyelocytic cells induced by alpha-helical peptides such as melittin, succinylated melittin and anionic peptides derived from the N-terminus of HA2 subunit of the influenza virus hemagglutinin, was measured. Melittin (GIGAVLKVLTTGLPALISWIKRKRQQ-NH2) caused a rapid (< 5 min) and dose-dependent (ED50 = 0.5 microM) permeabilization of HL60 cells at neutral pH, whereas the succinylated derivative induced cell permeabilization only at pH below 4.5 with an ED50 = 18 microM. The permeabilization by the anionic E5CA peptide (GLFEAIAEFIEGGWEGLIEGCA) containing 5 glutamic residues occurred (ED50 = 11 microM) at pH ranging from 6.5 to 6.0; replacing the tryptophan residue in position 14 by a phenylalanine residue decreased by about 1 unit the pH at which membrane permeabilization was effective. The membrane permeabilization activity of the E5CA peptide was reversibly abolished when the peptide was linked to a protein carrier. These results show that alpha-helical peptide-induced membrane permeabilization can be easily monitored by using flow cytometry in the presence of a non permeant dye. This method allows a rapid screening and an efficient mean of selection of peptides suitable to induce membrane permeabilization. PMID- 7488632 TI - Mammalian calcium-independent phospholipase A2. PMID- 7488634 TI - Reciprocal regulation of choline acetyltransferase and choline kinase in sympathetic neurons during cholinergic differentiation. AB - The regulation of the synthesis of acetylcholine and phosphatidylcholine in rat sympathetic neurons was examined in the context of cholinergic differentiation. We demonstrate that the activities of choline acetyltransferase (ChAT) and choline kinase (CK) are inversely affected by treatment of sympathetic neurons with retinoic acid, utilized as an agent that induces cholinergic differentiation. Whereas ChAT specific activity increased 2- to 4-fold after 12 days of treatment with 5 microM retinoic acid, CK specific activity decreased by 25-30%. These changes in enzyme activities were essentially reflected in the incorporation of [methyl-3H]choline into ACh and the metabolites of the CDP choline pathway for phosphatidylcholine synthesis. When sympathetic neurons were treated under high potassium conditions (50 mM) for 12 days, the specific activity of CK increased 1.3-fold whereas the activity of ChAT decreased by up to 90%. Furthermore, experiments in which the incorporation of [methyl-3H]choline into ACh and the metabolites of the CDP-choline pathway was measured in the absence of Na+ or in the presence of hemicholinium-3 (HC-3), demonstrate that CK has access to the same pool of choline utilized by ChAT. These results provide evidence that the activities of ChAT and CK may be inversely regulated during the process of cholinergic differentiation. PMID- 7488633 TI - Molecular heterogeneity of PAF in normal human mixed saliva: quantitative mass spectral analysis after direct derivatization of PAF with pentafluorobenzoic anhydride. AB - Platelet-activating factor (PAF), a family of phospholipid autacoids with potent pro-inflammatory activities, is present in saliva. The current study has quantitated various species of PAF isolated from normal human mixed saliva. Choline-containing, sn-2 acetylated phospholipids with sn-1 ether- or ester linked fatty alcohol/acid moieties (alkyl-PAF or acyl-PAF, respectively) were evaluated after direct derivatization with pentafluorobenzoic (PFB) anhydride. Individual species of PFB-derivatized PAF were separated by gas chromatography prior to mass spectral analysis; quantitative estimates of six different species of PAF in saliva were made by comparison to corresponding authentic, synthetic PAF standards. In each saliva sample, all six species of PAF were readily detected by this facile procedure. The predominant PAF was 1-O-hexadecyl-2-acetyl sn-glycero-3-phosphocholine or 16:0-alkyl-PAF (0.75 +/- 0.09 pmol/ml saliva; mean +/- S.E.; n = 5) which represented only 30.4 +/- 1.5% of the total PAF. Substantial amounts of 18:1- and 18:0-alkyl-PAF and 16:0-acyl-PAF were also identified (0.52 +/- 0.07, 0.35 +/- 0.06, and 0.35 +/- 0.02 pmol/ml saliva, respectively). In summary, mass spectrometric analysis of PAF after direct derivatization with PFB anhydride has revealed that at least six different species of PAF are present in normal human mixed saliva. This structural diversity may represent an important aspect of homeostasis in the healthy oral cavity. PMID- 7488635 TI - Heparin inhibits the accumulation of re-esterified cholesterol in macrophages loaded with acetylated low-density lipoprotein. AB - Heparin enhances the endocytosis of low density lipoprotein (LDL) in macrophages via a formation of complex with LDL. The direct effect of heparin on the metabolism of cholesterol in macrophages has not been elucidated. We therefore evaluated the effects of heparin on the accumulation and reesterification of cholesterol in cultured macrophages. We used acetylated LDL (acetyl-LDL), which lacks an affinity for heparin. Rat peritoneal macrophages induced with thioglycollate were incubated with 100 micrograms of acetyl-LDL for 14 h. Heparin significantly inhibited the accumulation of total and esterified cholesterol but did not affect the binding of 125I-labeled acetyl-LDL to macrophages or its cellular degradation. Heparin at concentration above 5 micrograms/ml inhibited the incorporation of [3H]oleate into cholesteryl oleate in macrophages. Heparin significantly inhibited the acyl CoA:cholesterol acyl transferase (ACAT) activity of macrophages by 68%. Data suggest that heparin inhibits the accumulation and reesterification of cholesterol in macrophages loaded with acetyl-LDL. Heparin like proteoglycans may thus protect the macrophages against the excessive accumulation of esterified cholesterol. PMID- 7488636 TI - Formation of pivaloylcarnitine in isolated rat heart cells. AB - Pivaloyl-containing antibiotics and pivalic acid in man or rat have been reported to cause increased urinary carnitine loss secondary to pivaloylcarnitine generation. Pivaloylcarnitine concentration was especially high in heart after administration of pivalic acid or pivampicillin in vivo. Formation of pivaloylcarnitine was therefore studied in isolated rat heart cells in the presence of sodium pivalate. Formation of pivaloylcarnitine in rat heart cells increased with incubation time, after a lag time from 0 to 2 h and linearly up to 6 h. The formation increased with increasing concentration of sodium pivalate, followed Michaelis-Menten kinetics with an apparent Km = 348 +/- 10 microM and Vmax = 116 +/- 20 pmol.mg protein-1.h-1. Bromoacetylcarnitine inhibited the pivaloylcarnitine formation to Ki = 116 +/- 43 microM and Vmax = 107 +/- 14 pmol.mg protein-1.h-1. The uptake of carnitine in heart cells was suppressed 62 74% by deoxycarnitine (40 microM) and D-carnitine (200 microM), and 95% by NaF (5 mM), NaN3 (500 microM) or at temperature 4 degrees C. Pivaloylcarnitine inhibited carnitine uptake to 33-35% of the controls, while sodium pivalate did not. More than 90% of intracellular pivaloylcarnitine was released from the heart cells after 18 h of incubation in the absence of sodium pivalate and L-carnitine. These data show that pivalate is readily converted to pivaloylcarnitine in heart cells, in contrast to the limited conversion in hepatocytes. PMID- 7488637 TI - Lung annexin II promotes fusion of isolated lamellar bodies with liposomes. AB - The role of annexin II in the secretion of lung surfactant was investigated using isolated lamellar bodies and/or liposomes as the model system for aggregation and fusion. We first compared membrane aggregation mediated by two forms of annexin II, annexin II monomer (Anx IIm) and annexin II tetramer (Anx IIt). Anx IIt required 20-fold less Ca2+ to mediate phosphatidylserine (PS) liposome aggregation compared to Anx IIm. Aggregation of lamellar bodies mediated by Anx IIt was 4-fold greater than that by Anx IIm at 1 mM Ca2+. These results suggest that Anx IIt may be the more active form in vivo. Fusion of lamellar bodies with PS liposomes was promoted by Anx IIt in a dose-dependent manner, with maximal fusion occurring at 10-15 micrograms/ml of Anx IIt. Fusion was dependent on Ca2+ and the phospholipid composition of liposomes. While the fusion of lamellar bodies with PS liposomes required 100 microM Ca2+, the fusion with PS/phosphatidylethanolamine (PE) (1:3) liposomes required only 10 microM Ca2+. Anx IIt-mediated lamellar body-liposome fusion was enhanced by arachidonic acid, a lung surfactant secretagogue and inhibited by 4.4'-diisothiocyanatostilbene 2,2'-disulfonic acid (DIDS), an inhibitor of lung surfactant secretion. The data suggest that Anx IIt may play a role in the fusion of lamellar bodies with plasma membranes during lung surfactant secretion. PMID- 7488638 TI - Anandamide hydroxylation by brain lipoxygenase:metabolite structures and potencies at the cannabinoid receptor. AB - Anandamide (arachidonyl ethanolamide) is a compound that was identified from porcine brain lipids by its ability to bind to the brain cannabinoid receptor. This study assessed anandamide as a substrate for a brain lipoxygenase and characterised the brain metabolite 12-hydroxyanandamide. Anandamide was also compared with arachidonic acid as a lipoxygenase substrate by examining enzyme kinetics in the presence of either of the two compounds. In addition, a non mammalian enzyme was used to generate 11- and 15-hydroxy-anandamide in order to compare the cannabinomimetic properties of a range of anandamide derivatives. A ligand displacement assay indicated a large variation in the affinity of anandamide metabolites for the brain cannabinoid receptor. The brain metabolite, 12-hydroxyanandamide had an affinity twice that of anandamide, although the 11- and 15- hydroxy-metabolites were considerably poorer ligands of this receptor. Consistent with the receptor binding data, 12-hydroxyanandamide (unlike 15 hydroxyanandamide) inhibited forskolin-stimulated cAMP synthesis, indicating it to be a functional agonist at the brain cannabinoid receptor. Pharmacological studies of the capacity of anandamide and its metabolites to inhibit the murine vas deferens twitch response indicated the 12-hydroxy-metabolite to be less active than the parent compound, but a better cannabinomimetic than 15 hydroxyanandamide. PMID- 7488639 TI - Direct observation of the action of cholesterol oxidase in monolayers. AB - The oxidation of monolayer cholesterol by cholesterol oxidase has been visualized using monolayer fluorescence microscopy. A direct microscopic visualization was possible because the lateral distribution of a lipid fluorophore, tetramethylrhodamine (TRITC)-labeled phosphatidylethanolamine, was very different in a cholesterol containing monolayer as compared with a cholestenone monolayer. The lipid fluorophore was effectively excluded from the condensed cholesterol phase, but was readily miscible in the cholestenone phase. One could therefore observe the appearance of fluorophore rich cholestenone-domains in the cholesterol monolayer as a result of the cholesterol oxidase catalyzed oxidation reaction. The oxidation experiments were performed at 22 degrees C with a monolayer surface pressure of 5 mN/m (on 50 mM Tris-HCl buffer, containing 140 mM NaCl, pH 7.4). When 40 mU/ml of cholesterol oxidase was injected beneath the monolayer under observation, it appeared that the enzyme penetrated the cholesterol monolayer at random sites and initiated the oxidation reaction. Once the oxidation reaction had commenced, it progressed rapidly and converted the condensed (cholesterol-rich) phase into an expanded (cholestenone-rich) phase. When the oxidation of cholesterol in mixed cholesterol/dimyristoylphosphatidyl choline monolayers was visualized, it was observed that the enzyme-catalyzed oxidation started from the expanded phases (domains with higher compressibility) and the reaction eventually led to the dissipation of the boundary line between expanded and condensed phases. With time all condensed phases were dissolved and the monolayer became uniformly fluorescent. The association of TRITC-labeled cholesterol oxidase with a non-fluorescent mixed cholesterol/dimyristoylphosphatidylcholine monolayer led to the penetration (or association) of the fluorescent cholesterol oxidase into expanded phases of the mixed monolayers. The monolayer lateral domain morphology was similar whether the fluorescent probe was TRITC-PE or TRITC-labeled enzyme. It is concluded that cholesterol oxidase associated with (or penetrated to some extent into) the expanded phases of a monolayer, and carried out its oxidation reaction in the expanded phase or at the interface between expanded and condensed phases. PMID- 7488640 TI - Evidence that lipid peroxidation products bind to DNA in liver cells. AB - Three lines of evidence are presented indicating the association of lipid peroxidation products with DNA in liver cells, labeled with [3H]arachidonic acid, in the presence of Fe(2+)-DTPA: (1) the nuclear DNA isolated from treated cells had higher radioactivity, compared to controls and the radioactivity increased with longer incubation times, (2) lipid-DNA adducts with a characteristic fluorescence spectrum were formed during the incubation with Fe(2+)-DTPA; (3) the association of peroxidation products with DNA could be inhibited by vitamin E and BHT. Compared with control DNA, purified lipid-DNA adducts showed a decrease of hyperchromicity and melting point, and partial resistance to hydrolysis by DNase I. On the other hand, the repair test shows that the lipid-DNA adducts in cells were not repaired by 4 h after removal of Fe(2+)-DTPA. A decrease in cell viability and in the activity of O6-alkylguanine acceptor protein was also observed with increasing incubation time. These data suggest that the lipid-DNA association, an oxidative DNA damage, occurs in cells treated by Fe(2+)-DTPA and could result in cytotoxicity if not repaired. PMID- 7488641 TI - The immunohistochemical localization of the non-specific lipid transfer protein (sterol carrier protein-2) in rat small intestine enterocytes. AB - A 13 kDa protein was isolated from rabbit small intestine brush-border membrane vesicles that was postulated to be involved in intestinal phosphatidylcholine (PC) and cholesterol uptake. This protein has cholesterol and PC-transfer activity in vitro (Turnhofer, H. et al. (1991) Biochim. Biophys. Acta 1064, 275 286) and has a molecular mass and isoelectric point similar to that of the non specific lipid transfer protein (nsL-TP, identical to sterol carrier protein-2). In addition, the first 28 N-terminal amino acid residues of the 13 kDa protein are nearly identical to nsL-TP from different species (Lipka, G. et al. (1995) J. Biol. Chem. 270, 5917-5925). In view of its possible role in intestinal lipid absorption, the localization of nsL-TP in rat small intestine was investigated using immunohistochemistry and immunoblotting. It is shown that nsLTP is predominantly localized in a subapical zone of the enterocyte but not in the brush-border membrane, thereby excluding a role in lipid uptake of this protein at the level of the plasma membrane. nsL-TP co-localized with the peroxisomal marker PMP70, underscoring earlier observations that nsL-TP is a peroxisomal protein. nsL-TP was found to be present along the entire length of the small intestine. The 58 kDa cross-reactive protein that was recently identified as a peroxisomal thiolase was shown to be present only in a small segment approximately halfway down the jejunum. The close apposition of the peroxisomes with the apical membrane and the discrete distribution of the 58 kDa protein may indicate that these organelles play a role in the intracellular processing of absorbed lipids. PMID- 7488643 TI - The morphologic change of endothelial cells by ancrod-generated fibrin is triggered by alpha v beta 3 integrin binding and the subsequent activation of a G protein coupled phospholipase C. AB - The mechanism of morphologic change of human cultured umbilical vein endothelial cells (HUVECs) caused by fibrin was investigated. Ancrod, a thrombin-like enzyme, did not cause morphologic alteration of HUVEC by itself at concentrations ranging from 0.01 to 10 U/ml. However, when 0.02 U/ml of ancrod was added to cultured HUVEC monolayers in the presence of citrated plasma, it caused pronounced morphologic change of HUVEC after 6-10 h incubation period. Gly-Pro-Arg-Pro (4 mg/ml), an inhibitor of fibrin polymerization, prevented the morphologic alteration, indicating that the morphologic alteration was caused by the polymerized fibrin. The morphologic change of HUVEC caused by ancrod-generated fibrin was not observed in the presence of an intracellular calcium mobilization inhibitor TMB-8 (50 microM), and the morphologic alteration was also less pronounced with BAPTA(15 microM)-loaded HUVECs and HUVECs pretreated with EGTA (1.2 mM). Ancrod (in Medium 199) itself did not stimulate phosphoinositide breakdown of HUVEC. However, when ancrod was present in plasma, it caused an increase of [3H]IP1 of HUVECs preloaded with [3H]myoinositol. This IP1 increment was inhibited by Gly-Pro-Arg-Pro. The increase of IP1 was significantly inhibited by the pretreatment of monoclonal antibodies 23C6 and 7E3 directed against alpha v beta 3 integrin. Neomycin (1 mM) and pertussis toxin (100 ng/ml), but not aspirin or mepacrine, blocked this enhanced phosphoinositide breakdown. The morphologic change was also prevented by the monoclonal antibodies, 23C6 and 7E3. These results suggest that both intra- and extra-cellular calcium participate in the event of morphologic change of HUVEC caused by ancrod-generated fibrin, and the morphologic change is mediated, at least in part, by fibrin binding to integrin alpha v beta 3 on HUVECs, causing the subsequent activation of the endogenous G-protein coupled phospholipase C. PMID- 7488642 TI - Identification of a mechanism of iron uptake by cells which is stimulated by hydroxyl radicals generated via the iron-catalysed Haber-Weiss reaction. AB - Recent studies have demonstrated that preincubation of SK-Mel-28 melanoma cells with ferric ammonium citrate (FAC) resulted in marked stimulation of 59Fe uptake from 59Fe-125I-transferrin (Tf), but only at Tf concentrations above that required for saturation of the Tf receptor (Richardson and Baker (1992) J. Biol. Chem. 267, 13972-13979). The mechanism responsible for this stimulation was unknown and is the subject of the present report. Preincubation of cells with FAC (25 micrograms/ml), followed by a 2 h incubation with 59Fe-125I-Tf (0.1 mg/ml; 1.25 microM), resulted in temperature-dependent 59Fe uptake to approx. 200% of the control value. Furthermore, the effect was not specific for melanoma cells and was also observed in other normal and neoplastic cells. Preincubation of melanoma cells with FAC also stimulated 59Fe uptake from 59Fe-citrate, but to a far greater extent than that observed with 59Fe-125I-Tf (viz., > 20-fold that seen for the control). Interestingly, neither receptor-mediated endocytosis nor the postulated diferric Tf reductase were involved in the FAC-activated Fe uptake process from Tf. However, the addition of free radical scavengers to FAC such as catalase, superoxide dismutase, ceruloplasmin, Hepes, mannitol and high concentrations of BSA or ascorbate, markedly depressed FAC-activated 59Fe uptake from 59Fe-125I-Tf and 59Fe-citrate. These agents when added to control cells had no effect on 59Fe uptake. The addition of superoxide generating agents and hydrogen peroxide to minimum essential medium (MEM) containing FAC but not to MEM alone, also stimulated 59Fe uptake. These data suggest that the initial activation of the FAC-stimulated Fe uptake system was caused by the production of hydroxyl radicals via the Fe-catalysed Haber-Weiss reaction. We propose that this Fe uptake process represents an important cellular defense mechanism against oxidant stress generated in the presence of low-molecular-weight Fe complexes. PMID- 7488644 TI - Effects of Na+/H+ antiport and intracellular pH in the regulation of HL-60 cell apoptosis. AB - The roles of Na+/H+ antiport and intracellular pH in apoptosis of HL-60 cells were investigated here. We found that dimethyl amiloride, a specific Na+/H+ antiport inhibitor, induced intracellular acidification but not apoptosis; while sodium ionophore, monensin caused intracellular alkalinization as well as apoptosis in HL-60 cells. Br-A23187 and thapsigargin could induce a various degree of intracellular alkalinization through the stimulation of Na+/H+ antiport. Dimethyl amiloride blocked the intracellular alkalinization and inhibited apoptosis induced by Br-A23187 and thapsigargin. PMA also stimulated Na+/H+ antiport and induced intracellular alkalinization which was completely blocked by dimethyl amiloride and partially attenuated by PKC inhibitors. PMA could inhibit apoptosis in HL-60 cells. PMA-induced suppression of apoptosis was, however, not interfered by dimethyl amiloride, but could be abolished by PKC inhibitors. These results indicate that pHi alkalinization and/or the stimulation of Na+/H+ antiport, instead of intracellular acidification, are contributory to the induction of apoptosis. PMA-induced inhibition of apoptosis is not necessarily associated with intracellular alkalinization, but primarily due to activation of PKC. We suggest that stimulation of Na+/H+ antiport and pHi alkalinization act as facilitating factors in the induction of apoptosis. PMID- 7488645 TI - Calcium signals in olfactory neurons. AB - Laser scanning confocal microscopy in combination with the fluorescent calcium indicators Fluo-3 and Fura-Red was employed to estimate the intracellular concentration of free calcium ions in individual olfactory receptor neurons and to monitor temporal and spatial changes in the Ca(2+)-level upon stimulation. The chemosensory cells responded to odorants with a significant increase in the calcium concentration, preferentially in the dendritic knob. Applying various stimulation paradigma, it was found that in a population of isolated cells, subsets of receptor neurons display distinct patterns of responsiveness. PMID- 7488646 TI - Interleukin-1 beta independently stimulates production of prostaglandin E2 and cyclic AMP from human decidual cells. AB - Interleukin-1 beta (IL-1 beta) increased the production of cyclic AMP and prostaglandin E2 (PGE2) by cultured human decidual cells during 24 h of stimulation, but not over short incubation times (< 6 h). At concentrations of IL 1 beta ranging from 1 to 100 pg/ml, there were parallel changes in cyclic AMP and PGE2 levels, but 1000 pg of IL-1 beta/ml inhibited cyclic AMP production while still stimulating PGE2 synthesis. The possible link between cyclic AMP and PGE2 was therefore studied further. Inhibition of IL-1 beta-stimulated PGE2 synthesis by indomethacin and direct addition of PGE2 had no effect on cyclic AMP levels, indicating that PGE2 did not increase cyclic AMP production by human decidual cells and confirming the independent synthesis of cyclic AMP and PGE2. The increase in cyclic AMP production induced by IL-1 beta is dependent on protein synthesis, but it is not known which component of the adenylate cyclase is increased. A phosphodiesterase inhibitor potentiated the effects of IL-1 beta on cyclic AMP synthesis, indicating that the cytokine may increase cyclic AMP metabolism. We suggest that high concentrations of IL-1 beta activate phosphodiesterase activity more than adenylate cyclase, which gives rise to the low levels of cyclic AMP noted above. IL-1 beta also decreased forskolin stimulated cyclic AMP production, which again indicates increased cyclic AMP metabolism. Since most concentrations of IL-1 beta alone increased cyclic AMP levels, this stimulation must out-weigh the increase in metabolism apparent in the presence of forskolin, phosphodiesterase inhibitor or high levels of interleukin. It is clear that IL-1 beta increased decidual PGE2 production independently of cyclic AMP, and that other second messenger must mediate the action of this cytokine. PMID- 7488647 TI - Deleterious effects of xanthine oxidase on rat liver endothelial cells after ischemia/reperfusion. AB - Previous studies have demonstrated that reactive oxygen species are involved in ischemic injury. The present work was undertaken to determine in vivo the role of xanthine oxidase in the oxygen free radical production during rat liver ischemia and to examine the activity of antioxidant enzymes (superoxide dismutase, catalase and glutathione peroxidase) during the same period. Our results indicate a 4-fold increase in xanthine oxidase activity between 2 and 3 hours of normothermic ischemia, in parallel with a decrease in cell viability. Moderate hypothermia delays both events. Under the same conditions, the activity of oxygen radical scavenging enzymes remains unchanged. Moreover, we have compared in vitro the susceptibility of isolated liver cells to an oxidative stress induced by O2. , H2O2 and .OH. Our results reveal that endothelial cells are much more susceptible to reactive oxygen species than hepatocytes, probably because they lack H2O2-detoxifying enzymes. These findings suggest that xanthine oxidase might play a major role in the ischemic injury mainly at the level of the sinusoidal space where most endothelial cells are located. PMID- 7488648 TI - Modulating hypoxia-induced hepatocyte injury by affecting intracellular redox state. AB - Hypoxia-induced hepatocyte injury results not only from ATP depletion but also from reductive stress and oxygen activation. Thus the NADH/NAD+ ratio was markedly increased in isolated hepatocytes maintained under 95% N2/5% CO2 in Krebs-Henseleit buffer well before plasma membrane disruption occurred. Glycolytic nutrients fructose, dihydroxyacetone or glyceraldehyde prevented cytotoxicity, restored the NADH/NAD+ ratio, and prevented complete ATP depletion. However, the NADH generating nutrients sorbitol, xylitol, glycerol and beta hydroxybutyrate enhanced hypoxic cytotoxicity even though ATP depletion was not affected. On the other hand, NADH oxidising metabolic intermediates oxaloacetate or acetoacetate prevented hypoxic cytotoxicity but did not affect ATP depletion. Restoring the cellular NADH/NAD+ ratio with the artificial electron acceptors dichlorophenolindophenol and Methylene blue also prevented hypoxic injury and partly restored ATP levels. Ethanol which further increased the cellular NADH/NAD+ ratio increased by hypoxia also markedly increased toxicity whereas acetaldehyde which restored the normal cellular NADH/NAD+ ratio, prevented toxicity even though hypoxia induced ATP depletion was little affected by ethanol or acetaldehyde. The viability of hypoxic hepatocytes is therefore more dependent on the maintenance of normal redox homeostasis than ATP levels. GSH may buffer these redox changes as hypoxia caused cell injury much sooner with GSH depleted hepatocytes. Hypoxia also caused an intracellular release of free iron and cytotoxicity was prevented by desferoxamine. Furthermore, increasing the cellular NADH/NAD+ ratio markedly increased the intracellular release of iron. Hypoxia induced hepatocyte injury was also prevented by oxypurinol, a xanthine oxidase inhibitor. Polyphenolic antioxidants or the superoxide dismutase mimic, TEMPO partly prevented cytotoxicity suggesting that reactive oxygen species contributed to the cytotoxicity. The above results suggests that hypoxia induced hepatocyte injury results from sustained reductive stress and oxygen activation. PMID- 7488649 TI - Khellin, but not 8-methoxypsoralen, inhibits adenylyl cyclase system in HeLa cells. AB - Until recently, the therapeutic effects of furocoumarins and furochromones plus UV-A light were thought to be due to their ability to form photoadducts with DNA in the cell nuclei; now it appears that membrane effector systems may be involved as targets. Here we show that in HeLa cells khellin at 1 and 5 microM final concentration, in combination with UV-A light, inhibits NaF-stimulated adenylyl cyclase activity and Pertussis Toxin (PT)-catalyzed ADP-ribosylation of alpha subunits of inhibitory guanine nucleotide regulatory proteins (Gi) and increases GTPase activity. In the same experimental conditions, 8-methoxypsoralen (8-MOP), either alone or plus UV-A, does not affect adenylyl cyclase and GTPase activities. Our results suggest that in HeLa cells, through an interaction with a receptor and the mediation of Gi proteins, the adenylyl cyclase system is a target for khellin but not for 8-MOP. PMID- 7488651 TI - The transport of acidic amino acids and their analogues across monolayers of human intestinal absorptive (Caco-2) cells in vitro. AB - The X-AG system, a sodium-dependent, acidic amino-acid transport system has been implicated in the transport of L-aspartate and L-glutamate across monolayers of human Caco-2 cells, an in vitro model of intestinal absorption. This system, which shares many properties with the L-glutamate carrier present in the human jejunum, is highly saturable (> 95% at 50 microM), vectorial (apical-to basolateral >> basolateral-to-apical) and sodium-, pH- and temperature-dependent. L-Aspartate was also transported against a 10-fold reverse concentration gradient. These data are consistent with a major (saturable) carrier-mediated pathway superimposed onto a minor non-saturable (diffusional) pathway. The carrier has an absolute sodium-dependence and the Michaelis constants for the sodium-dependent transport component (Km) for L-aspartate and L-glutamate were 56 +/- 3 microM and 65 +/- 6 microM, respectively. Cross-inhibition studies showed that strong interaction with the carrier was limited to close analogues of the natural substrates. Potent inhibitors included L-aspartate, D-aspartate (Ki, 70 microM), L-glutamate (Ki 180 microM) and threo-beta-hydroxy-DL-aspartate (Ki, 55 microM), while partial inhibitors included alpha-methyl-DL-aspartate, D glutamate, L-asparagine, L-proline and L-alanine. Replacement of the side-chain COO- group (aspartate) with -SO-3 (L-cysteate, Ki, 65 microM) or -(H)P(O)O- (DL-3 (hydroxyphosphoryl)alanine, Ki, 60 microM) maintained strong interaction with the carrier while -As(O)(OH)O- (DL-3-arsonoalanine, Ki, 1100 microM) and -P(O)(OH)O- (DL-3-phosphonoalanine, Ki, 3270 microM) were much more weakly bound, with the larger, but probably less ionised, arsono analogue being more tightly bound than the phosphono compound. The corresponding analogues of glutamate (homologous extension of the methylene chain) showed negligible interaction. We conclude that Caco-2 monolayers are a relevant experimental model for the study of the transport of acidic amino acids and their analogues in man. PMID- 7488652 TI - Protein phosphorylation in apoptosis. PMID- 7488653 TI - Three-dimensional display of cardiac structures using reconstructed magnetic resonance imaging. AB - It is sometimes difficult to understand the three-dimensional (3D) relationship of cardiac and mediastinal structures despite advances in magnetic resonance (MR) imaging techniques. We present a low-cost system for 3D reconstruction of the major mediastinal structures by processing the MR imaging data on a NeXT workstation. MR images of multisection, multiphase, spin-echo techniques stored in a picture archiving and communication system (PACS) data base were used for the reconstruction. The computer program obtained the contours of the multiple components of the mediastinal structures by the combination of automatic and manual procedure. The bundled software of a 3D kit was used for surface rendering of hidden surface removal, shading of the visible parts of the surfaces, perspective transformation, and motion parallax by rotation of the surfaces. 3D reconstruction was performed in 15 patients with cardiac diseases, and the 3D reconstructed images were compared with the plain chest x rays of the patients. The 3D presentation clearly showed the complex anatomy of cardiovascular diseases and helped elucidate the misconceptions in the interpretation of the plain chest x rays. Our 3D images are used for education and should be viewed by medical students and beginners in radiology at an individual pace with plain chest radiographs, MR images, and legends. Although applied to the heart and the great vessels in this report, this system is also applicable to other structures. PMID- 7488650 TI - Differential expression of thromboxane A synthase and prostaglandin H synthase in megakaryocytic cell line. AB - We determined the expression of isoforms of prostaglandin H synthase (PGHS) and thromboxane A synthase (TXAS) in a human megakaryocyte cell line (MEG-01. The basal levels of full-length TXAS mRNA and the 60 kDa TXAS protein were high when compared to those of PGHS-1 and PGHS-2 in uninduced cells. Despite a high TXAS level, uninduced MEG-01 cells synthesized only a small amount of thromboxane A2 (TXA2) due to limited PGHS-1 or PGHS-2 expressions. Following PMA induction there was little change in TXAS. PGHS-2 mRNA was significantly increased at only 3 h of PMA treatment and the level declined rapidly, whereas PGHS-1 mRNA and protein levels were concordantly stimulated. Induction of PGHS-1 reached plateau on day 3 of PMA treatment. Analysis of arachidonate metabolism in cells induced by PMA for 3 and 5 days showed a high level of PGH2 synthesis which exceeded the TXAS capacity for TXA2 synthesis. Only traces of PGHS-2 mRNA and alternate-spliced TXAS mRNA were detected in human platelets. We conclude that TXAS and PGHS are differentially expressed in MEG-01 during PMA-induced differentiation. PMID- 7488654 TI - Neural network reconstruction of single-photon emission computed tomography images. AB - An artificial neural network (ANN) trained on high-quality medical tomograms or phantom images may be able to learn the planar data-to-tomographic image relationship with very high precision. As a result, a properly trained ANN can produce comparably accurate image reconstruction without the high computational cost inherent in some traditional reconstruction techniques. We have previously shown that a standard backpropagation neural network can be trained to reconstruct sections of single photon emission computed tomography (SPECT) images based on the planar image projections as inputs. In this study, we present a method of deriving activation functions for a backpropagation ANN that make it readily trainable for full SPECT image reconstruction. The activation functions used for this work are based on the estimated probability density functions (PDFs) of the ANN training set data. The statistically tailored ANN and the standard sigmoidal backpropagation ANN methods are compared both in terms of their trainability and generalization ability. The results presented show that a statistically tailored ANN can reconstruct novel tomographic images of a quality comparable with that of the images used to train the network. Ultimately, an adequately trained ANN should be able to properly compensate for physical photon transport effects, background noise, and artifacts while reconstructing the tomographic image. PMID- 7488655 TI - Opinion: a prototype for a computerized national mammography registry and tracking system using telecommunications and the Internet. AB - In this report, we analyze the feasability and discuss the potential benefits of using currently available technology for the wide-area registration and tracking of mammography patients. In our prototype, three dissimilar computer systems transmitted mammographic data (demographics and the results of mammograms) in a standardized format to a central data repository. Two of the three systems were dedicated computerized mammography systems and one was a general-purpose radiology information system. High-speed modems and the Internet were used to connect with the central repository, which could be queried in real time by remote users. Our results indicated that a busy mammography practice, using the slowest transmission method we tested (14-kilobaud modem), could transmit several days of mammographic data to a central repository in a matter of minutes. To implement systems that provide nation-wide mammographic tracking and follow up, more in-depth planning, development, and testing are necessary. PMID- 7488656 TI - Differentiation between nodules and end-on vessels using a convolution neural network architecture. AB - In recent years, many computer-aided diagnosis schemes have been proposed to assist radiologists in detecting lung nodules. The research efforts have been aimed at increasing the sensitivity while decreasing the false-positive detections on digital chest radiographs. Among the problems of reducing the number of false positives, the differentiation between nodules and end-on vessels is one of the most challenging tasks performed by computer. Most investigators have used a conventional two-stage pattern recognition approach, ie, feature extraction followed by feature classification. The performance of this approach depends totally on good feature definition in the feature extraction stage. Unfortunately, suitable feature definition and corresponding extraction implementation algorithms proved to be very difficult to define and specify. A convolution neural network (CNN) architecture, trained by direct connection to the raw image is proposed to tackle the problem. The CNN, which uses locally responsive activation function, is directly and locally connected to the raw image. The performance of the CNN is evaluated in comparison to an expert radiologist. We used the receiver operating characteristics (ROC) method with area under the curve (Az) as the performance index to evaluate all the simulation results. The CNN showed superior performance (Az = 0.99) to the radiologist's (Az = 0.83). The CNN approach can potentially be applied to other applications, such as the differentiation of film defects and microcalcifications in mammography, in which the image features are difficult to define or not known a priori. PMID- 7488657 TI - Threshold selection for CT angiography shaded surface display of the renal arteries. AB - The purpose of this study was to define objective criteria to calculate a tissue segmentation threshold for shaded surface display (SSD) rendering of the renal arteries with computed tomography angiography. Contrast-enhanced spiral CT scans were obtained through the renal arteries of nine patients. Six sets of SSD images were rendered for each patient with lower threshold values ranging from 80 to 130 Hounsfield units (HU) by increments of 10 HU. Visceral organ enhancement was measured in the aorta, liver, spleen, pancreas, and kidney. The segmentation threshold for each patient was determined by evaluation of the SSD images alone as well as by comparison with conventional arteriograms. The ideal threshold, as shown by comparison with conventional arteriography, was better correlated with a threshold value selected by qualitative evaluation of SSD images alone (rs = .42), than with measured enhancement in visceral organs (rs = -.289 to .009). The degree of stenosis was overestimated in a single renal artery (1 of 18) because of an inappropriate threshold selected by evaluation of the SSD images alone. In comparison with a segmentation threshold calculated from measured enhancement of visceral organs, a segmentation threshold selected by qualitative evaluation of the resulting SSD images is more likely to approximate the ideal threshold. Given the subjective nature of such threshold selection, further evaluation is warranted to determine whether threshold selection may result in inaccurate grading of stenosis. PMID- 7488658 TI - Blunt splenic injury in adults: a decision analysis comparing options for treatment. AB - OBJECTIVE: To evaluate factors which influence surgeons in their decision to treat blunt splenic injuries in adults by observation, splenorrhaphy or splenectomy. DESIGN: Decision analysis. SETTING: Army hospital, USA. MATERIAL: 56 Selected papers on the treatment of splenic trauma published between 1983 and 1993. INTERVENTION: Variables evaluated included the incidence of transfusion, postsplenectomy sepsis, posttransfusion hepatitis, chronic active hepatitis, cirrhosis, and rebleeding. MAIN OUTCOME MEASURES: Quality adjusted life expectancies (QALE) after each procedure. RESULTS: QALEs when the mean incidence of each variable was used in the decision analysis were 39.68 years for observation, 39.56 years for splenorrhaphy, and 38.94 years for splenectomy. Sensitivity analysis showed that when the incidence of transfusion was controlled, splenorrhaphy was superior, and both splenorrhaphy and observation were superior to splenectomy. Once a patient had been given blood products the advantages of non-operative management were lost. CONCLUSIONS: Only patients with minor splenic injuries who did not require blood transfusion should be treated by observation. Patients who require blood transfusion should be explored with the object of salvaging the spleen. Some patients with severe, devitalising injuries will require splenectomy despite the best attempts to salvage the organ. PMID- 7488659 TI - Tuberculosis of the breast. AB - OBJECTIVE: To present our 20 years experience of tuberculosis of the breast. DESIGN: Retrospective study. SETTING: Teaching hospital, Turkey. SUBJECTS: 9 women with tuberculous mastitis. MAIN OUTCOME MEASURE: Cure. RESULTS: All cases underwent frozen section and excision. One required a simple mastectomy because of the extent of destruction; the remainder underwent lumpectomy. All patients were given rifampicin, ethambutol, and isoniazid, and the three who had tuberculosis of other organs were also treated with streptomycin. Mean follow up was 87 months (range 6-178) and two patients were lost to follow up, at three and six years, respectively. Histological examination showed the presence of tubercle and central caseation in 8 cases and granulomatous infiltration in one. CONCLUSION: Tuberculous mastitis is rare, and should be suspected in any woman with persistent breast abscesses and sinuses, particularly if she lives in an area from which tuberculosis has not been eradicated. Conservative surgery and antituberculous drugs are the treatment of choice. PMID- 7488660 TI - The effects of transplantation of hepatocytes cultured with insulin on acute liver failure induced by 90% hepatectomy in the rat. AB - OBJECTIVE: To evaluate the influence of transplantation of hepatocytes either not cultured or cultured with or without insulin on survival in rats with acute liver failure induced by 90% hepatectomy. DESIGN: Randomized laboratory experiment. SETTING: University department, Sweden. MATERIAL: 60 Adult male Sprague-Dawley rats, not including donors. INTERVENTIONS: 90% hepatectomy without hepatocyte transplantation, or with intrasplenic transplantation of: uncultured hepatocytes immediately after resection, or uncultured hepatocytes one or three days before resection, or hepatocytes cultured with or without insulin immediately after resection (n = 10 in each group). MAIN OUTCOME MEASURES: Cell viability, attachment ability, survival, liver function, and body weight. RESULTS: Transplantation of hepatocytes cultured for 3 days without insulin or of uncultured hepatocytes one day prior to hepatectomy did not significantly improve 30-day survival compared with uncultured hepatocyte transplantation immediately after hepatectomy. Hepatocytes cultured with insulin for 3 days and transplanted immediately after hepatectomy significantly improved survival compared with animals in which hepatocytes were transplanted either uncultured or cultured for 3 days without insulin immediately after hepatectomy. Hepatocyte transplantation 3 days before hepatectomy further increased survival compared with the other groups in which transplantation was done immediately after hepatectomy. Acute failure, acute compensatory, and late 'enhanced' phases were defined within 30 days of hepatectomy. CONCLUSION: Transplantation of hepatocytes that are cultured with hepatocellular trophic factors (for example insulin) before transplantation may have the potential to increase the effectiveness of transplanted hepatocytes in the treatment of acute liver failure. PMID- 7488661 TI - Influence of white cell count on surgical decision making in patients with abdominal pain in the right lower quadrant. AB - OBJECTIVE: To evaluate the influence of white cell count (WCC) on the decision to operate on patients with right lower abdominal pain. DESIGN: Open study. SETTING: University hospital, Switzerland. SUBJECTS: 221 adult patients admitted to emergency department with right lower abdominal pain. INTERVENTIONS: Surgeons were unaware of the WCC until after they had initially decided whether to remove the appendix. They were told the result and asked if it would influence their final decision. MAIN OUTCOME MEASURE: Whether surgeons changed their minds about operation when told the initial WCC. RESULTS: 112 Patients (51 men and 61 women) were operated on, of which 44 men (52%) and 42 women (31%) had histologically confirmed appendicitis. The decision to operate was influenced by the initial WCC in only 6 patients (3%, 95% confidence interval (CI) 0.6 to 4.9%). At follow up after 30 days the change of decision in the 6 patients had been correct in 5. CONCLUSION: The WCC did not significantly influence surgical decision-making in the group of patients who were suspected of having acute appendicitis. PMID- 7488662 TI - Impact of a programme of autologous blood donation on the incidence of infection in patients with colorectal cancer. AB - OBJECTIVE: To assess the effects of both autologous and homologous blood transfusion on the incidence of infective complications after elective operations for colorectal cancer. DESIGN: Prospective open study. SETTING: University hospital, Italy. INTERVENTIONS: Recording of variables known to influence the development of infection. MAIN OUTCOME MEASURES: Infective morbidity and mortality. RESULTS: Fifty-three patients (33%) deposited their own blood. Eighty six of the 161 (53%) patients were transfused, 36 were given autologous blood, 48 homologous blood and 2 both autologous and homologous blood. These two were excluded from the analysis. Infective complications developed in 28 patients (17%), of which 7/75 (9%) were in patients who had not been transfused, 5/56 (14%) in patients given autologous transfusion, and 16/48 (33%) in patients given homologous transfusions (p < 0.001). Multivariate analysis identified homologous blood transfusion as the only variable significantly associated with the development of postoperative infective complications. No patient died. CONCLUSION: Transfusion of autologous blood was associated with significantly fewer postoperative infective complications than transfusion of homologous blood or no blood transfusion. PMID- 7488663 TI - Alterations in lymphocyte subsets as prognosticators of postoperative infections. AB - OBJECTIVE: To evaluate changes in lymphocyte subsets after major abdominal and thoracic operations, and to correlate changes with the development of clinically relevant infections postoperatively. DESIGN: Open study. SETTING: University hospital, Italy. SUBJECTS: 33 patients who were to undergo major abdominal or thoracic operations. INTERVENTIONS: Lymphocyte subsets were measured by cytofluorimetry before operation and 1, 3, 5, and 7 days postoperatively. MAIN OUTCOME MEASURES: Correlation between changes in the number of lymphocyte subsets and development of infection. RESULTS: Lymphocyte subsets were within the reference range in all patients before operation. 10/33 Patients developed infections (pneumonia, bacteraemia, or wound or urinary tract infections) between the second and the ninth days postoperatively (30%). On day 1 the numbers of all lymphocyte subsets had decreased significantly compared with the preoperative measurements in all patients (CD3 p < 0.01, CD4 p < 0.001, and CD8 p < 0.05). The reduction in CD3 was significantly greater in the group that developed infections (p < 0.001). Among patients who did not develop infections the numbers of lymphocyte subsets had returned to the reference range within a week of operation whereas among patients who developed infections they remained depressed (p < 0.05). CONCLUSION: The synchronous reduction in numbers of all lymphocyte subsets on the first day postoperatively to below 50% of the reference range (CD3 to < 600/microliters, CD4 to < 400/microliters, and CD8 to < 250/microliters) predicted the development of infection postoperatively with an accuracy of 89%, a sensitivity of 80%, and a specificity of 96%. PMID- 7488665 TI - Infection after laparoscopic cholecystectomy: are antibiotics really necessary? AB - OBJECTIVE: To establish the incidence of infection after laparoscopic cholecystectomy, and assess the need for antibiotic prophylaxis. DESIGN: Prospective open study. SETTING: University teaching hospital, United Kingdom. SUBJECTS: 253 consecutive patients undergoing laparoscopic cholecystectomy between September 1990 and January 1993. INTERVENTIONS: A single intravenous dose of cefuroxime 1.5 g at induction of general anaesthesia. MAIN OUTCOME MEASURES: Infective complications. RESULTS: Patients were reviewed at two weeks and 12 months. At two weeks there had been two wound infections (one resolved spontaneously and the other required removal of a gallstone from the subcutaneous tissue), two chest infections (treated with antibiotics orally and physiotherapy), and one subhepatic abscess (drained percutaneously under ultrasonographic control). No other complications were reported at 12 months. CONCLUSIONS: Routine antibiotic prophylaxis may be unnecessary during elective laparoscopic cholecystectomy, but a randomised controlled trial is necessary to confirm this. PMID- 7488664 TI - Intraperitoneal micro-organisms and the severity of peritonitis. AB - OBJECTIVE: To find out if there was any correlation between the type or number of pathogenic bacteria in peritoneal exudate, the values of various prognostic scores, the inflammatory response, and the outcome, in patients with peritonitis. DESIGN: Prospective open study. SETTING: University hospital, Germany. SUBJECTS: 51 Consecutive patients with secondary peritonitis. INTERVENTIONS: Laparotomy within 12 hours of admission or within 8 hours of diagnosis in 9 patients with postoperative peritonitis. MAIN OUTCOME MEASURES: Correlation between the severity of the disease (APACHE II score, Septic Severity Score, Mannheim Peritonitis Index, Peritonitis Index Altona II, and outcome), the intraperitoneal and the systemic inflammatory response, intraperitoneal and systemic endotoxin concentrations, and type and number of micro-organisms grown from peritoneal fluid. RESULTS: Intra-abdominal microbiological findings did not correlate with severity of illness judged by the scoring systems, the later incidence of infective complications, or the final outcome. The presence of intra-abdominal microbes was associated with signs of a systemic inflammatory response (median activation index 3 (range 1-3) compared with 2 (range 1-3)), the length of history (median 52 hours (range 3-72) compared with 16 hours (range 3-56), and local and systemic concentrations of endotoxin (peritoneal exudate: median 4800 EU/ml (range 0.06-136674) compared with 220 EU/ml (range 0.00-1800); plasma: median 0.05 EU/ml (range 0.00-1.32) compared with 0.04 EU/ml (range 0.00-0.13)). The sensitivity of the organisms to the antibiotics given (cefuroxime and metronidazole) did not influence the incidence of later infective complications or the outcome. CONCLUSIONS: These results do not suggest micro-organisms invading the peritoneal cavity and respective antibiotic treatment to be major determinants of the clinical course of peritonitis. PMID- 7488666 TI - Fibre is an essential ingredient of enteral diets to limit bacterial translocation in rats. AB - OBJECTIVE: To assess the effect of six different enteral diets on the gut barrier. DESIGN: Laboratory study. SETTING: University hospital, Germany. MATERIAL: 70 Specific pathogen free female Crl:CDR BR rats. INTERVENTIONS: For 7 days, 6 groups of rats were fed orally with standard chow (n = 15); total parenteral nutrition solution (oral TPN, n = 15); elemental diet (ED, n = 10); nutrient-defined diet (NDD, n = 10); or the NDD supplemented with uracil (NDD+uracil, n = 10), or fibre (NDD+fibre, n = 10). MAIN OUTCOME MEASURES: Bacterial translocation to mesenteric lymph nodes, numbers of Gram negative enterobacteria and total aerobic bacteria in the caecum, and intestinal concentrations of secretory IgA. RESULTS: The incidence of bacterial translocation was significantly increased in the groups given oral TPN, ED, NDD, and NDD+uracil compared with the group given chow. Only NDD+fibre resulted in a similar degree of translocation to that in the chow group. All groups in which there was increased translocation had a highly significant overgrowth of aerobic bacteria in the caecum, mainly by Gram negative enteric organisms. The secretory IgA concentration was reduced in the group that had been given oral TPN, and that in the ED and NDD+uracil groups was similar to that in the chow group. NDD and NDD+fibre were associated with higher intestinal concentrations of secretory IgA than chow. CONCLUSION: Fibre-free enteral diets do not protect the gut antimicrobial barrier whatever else is in them. The superiority of early enteral as opposed to parenteral nutrition after injury may, therefore, not be the result of a specific protective effect on the gut barrier. The supplementation of commercial enteral diets with bulk fibre should be tested in clinical trials. PMID- 7488667 TI - Vein patch rupture: early complication of carotid endarterectomy. PMID- 7488668 TI - Isolated oesophageal tuberculosis: a rare cause of oesophageal stricture. PMID- 7488669 TI - Primary dissection of the abdominal aorta. PMID- 7488670 TI - Ultrasound diagnosis of incarcerated obturator hernia. PMID- 7488671 TI - Peritoneal herniation and intestinal obstruction: a complication of laparoscopic inguinal herniorrhapy. PMID- 7488673 TI - Proliferation of mature and immature subpopulations of bronchoalveolar monocytes/macrophages and peripheral blood monocytes. AB - A continuous influx of peripheral blood monocytes (PBM) to the lung is thought to maintain the local population of alveolar macrophages (AM). However, local proliferation of a small subpopulation of AM has been demonstrated in animal studies and in humans. AM exhibit a great heterogeneity with regard to their morphology (cell size, shape of nucleus), immunophenotype (expression of CD14 and RFD9 antigen), and function. Part of this heterogeneity may be explained by the presence of different maturation stages of AM, ranging from small immature, CD14+ RFD9- PBM-like cells to large, CD14- RFD9+ mature AM. These findings prompted us to study whether proliferation of PBM and AM is related to their stage of maturation. The expression of the proliferation marker Ki-67 was studied in AM from both healthy volunteers and patients suffering from sarcoidosis. Using double immunofluorescence staining, we studied proliferation of immature, CD14+ AM, and mature, RFD9+ AM in sarcoidosis, and we compared this with PBM. A significantly larger percentage of AM in general expressed Ki-67 antigen in sarcoidosis (3.0 (median); range 1.1-5.5) as compared with healthy volunteers (0.8; 0.2-1.3). In sarcoidosis, proliferation was observed in both the immature and the mature subpopulation of AM. Proliferating PBM were rarely observed [less than 0.2% of the CD14+ mononuclear cells (MNC)] both in healthy volunteers and sarcoidosis patients. A small subpopulation of PBM showed a weak expression of RFD9 antigen (less than 1% of MNC). Interestingly, proliferation of PBM was concentrated in this subpopulation (15% of the RFD9+ MNC). These data show that even mature AM, which are generally thought to be terminally differentiated cells with little capacity to replicate, are able to proliferate, whereas a relatively very low percentage of their precursors in the blood circulation proliferates. Furthermore, the findings suggest that lung tissue in sarcoidosis creates an environment which promotes proliferation of monocytic cells. Pulmonary alveolar macrophages (AM) were originally recognized as phagocytosing scavenger cells (Ham & Cormack 1979), but presently they are also known to initiate and regulate inflammatory and immunological processes in several lung diseases (Herscowitz 1985, Unanue & Allen 1987, Sibille & Reynolds 1990). AM are thought to represent more mature cells of the mononuclear phagocyte system, and to be derived from peripheral blood monocytes (PBM) (Van Furth 1982, Ginsel 1993). As AM are continuously lost (mainly through a transport from the peripheral airways, via the trachea to the pharynx), the local AM population must be constantly replenished.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 7488672 TI - Effects of omeprazole on cell kinetics of carcinogen-induced colon tumours in rats. AB - This study was designed to evaluate the effects of hypergastrinaemia induced via suppression of gastric acid by omeprazole on carcinogen-induced colon cancer in rats. The carcinogen methylazoxymethanol (MAM), 30 mg/kg, was administered intraperitoneally at 6-weekly intervals to Sprague-Dawley rats. Four weeks after the last MAM injection, the first daily dose of omeprazole, 40 mg/kg, was given by gastric gavage to one group of rats, and the rest were given buffered methylcellulose vehicle. After 10 weeks of daily omeprazole or vehicle, the rats were anaesthetized with ether, blood samples obtained, and animals sacrificed. Gastrin levels in serum from omeprazole-treated rats were elevated nearly six fold. DNA and RNA levels in gastric mucosa were unchanged by omeprazole, but protein content was somewhat reduced. No biochemical or histological changes related to omeprazole treatment were observed in normal colon. The number of tumours, tumour volumes, and total tumour burden were not significantly different in colons of vehicle- or omeprazole-treated rats. Analysis by flow cytometry revealed that the S phase fraction was lower in tumour cells from omeprazole treated animals; and that the frequency of DNA aneuploidy was also reduced. The results indicate that while omeprazole-induced suppression of stomach acid in rats elevates levels of gastrin in serum, it does not substantially alter the biochemical or cellular characteristics of carcinogen-induced colon tumours. PMID- 7488674 TI - A stochastic model for dual label experiments: an analysis of the heterogeneity of S phase duration. AB - We develop a statistical approach for the study of S phase duration in experiments using sequential pulses of two thymidine analogues. Cell entrance into S phase is assumed to follow a possibly nonhomogeneous Poisson process, and the duration in S phase independently follows an unspecified distribution, thus allowing the possibility of variable S phase duration times for cells. Several conclusions regarding experimental design considerations are reached. The availability of three labelled cell subgroups comprising cells receiving exactly one of the two thymidine analogues or receiving both thymidine analogues at least doubles the efficiency of the mean S phase duration estimate compared to conventional estimates based on two labelled groups. Increasing the duration between the two thymidine analogue pulses can also dramatically increase the efficiency. The modelling technique was applied to fourteen uveal melanomas from patients who received in vivo injections of two thymidine analogues, bromodeoxyuridine (BrdUrd) and iododeoxyuridine (IdUrd). Counts of labelled cells were consistent with steady-state time homogeneous entry into S phase and nonhomogeneous spatial entry into S phase. PMID- 7488675 TI - Adverse effects of potential agents for the treatment of Alzheimer's disease: a review. PMID- 7488676 TI - Investigations into the toxic effects of alcohol on skeletal muscle. PMID- 7488677 TI - Dr Thomas Young--polymath. PMID- 7488678 TI - Long term neurotoxic effects of chemical warfare organophosphate compounds (Sarin) PMID- 7488679 TI - Long term neurotoxic effects of organophosphate compounds. PMID- 7488681 TI - [Training of family and community medicine specialists in emergency care of patients]. PMID- 7488680 TI - [Future of general/family medicine in Spain and possible role of the Spanish Society of Family and Community Medicine]. PMID- 7488683 TI - [Variables related to the self-perceived health status: a population study]. AB - OBJECTIVE: To study the self-perceived health status (PHS) of a community and its relationship to socio-economic variables of morbidity and frequency of attendance at the doctor's. DESIGN: A descriptive, crossover study. SETTING: A base Health District (HD). PATIENTS AND OTHER PARTICIPANTS: The general catchment area for the Occidente HD, Cordoba (N = 12,801 inhabitants) provided a random sample (for a p = 0.5, alpha error = 0.05 and with precision - 5%) of individuals over 15 (N = 384). MEASUREMENTS AND MAIN RESULTS: By means of a survey with a personal interview at home, socio-demographic data, PHS, chronic morbidity, consumption of pharmaceuticals and attendance at the doctor's were gathered. 32.9% (C.I. 95%: 27.4-37.1) stated a negative PHS. It was confirmed that older age (O.R. = 1.6; C.I. 1.04 - 1.08) and the existence of an acknowledged chronic pathology (O.R. = 6.84); C.I. 2.0 - 23.33) were the variables most strongly associated to a negative PHS. Chronic pathologies which were most intensely related to an unfavorable PHS were: Bronchitis (O.R. = 8.08; C.I. 3.02 - 21.5), Diabetes Mellitus (O.R. = 6.35; C.I., 1.03 - 38.9), Arthrosis (O.R. = 3.53; C.I. 2.03 - 6.13), Depression (O.R. = 3.53; C.I. 1.79 - 6.96), Arterial Hypertension (O.R. = 3.52; C.I. 1.63 - 7.63) and peptic ulcer (O.R. = 2.05; C.I. 1.02 - 4.11). CONCLUSIONS: The PHS could be useful as an overall indicator of the level of health of the population, as it is easy to obtain and has been shown to be linked to explanatory variables such as age and the presence of chronic disorders. PMID- 7488685 TI - [Primary care research in the Valencia community. Original articles published over a 4-year period (1990-1993)]. AB - OBJECTIVE: To evaluate the quality of Primary Care (PC) research in the Community of Valencia through research articles. DESIGN: Descriptive study. SETTING: Primary Care in the community of Valencia. PATIENTS AND OTHER PARTICIPANTS: Research articles on PC published 1990-1993 by professionals from the Community of Valencia in: Atencion Primaria, Medicina Clinica, Revista Clinica Espanola and Gaceta Sanitaria. MEASUREMENTS AND MAIN RESULTS: 43 articles published between 1990 and 1993 (6, 6, 9 and 22, respectively) were obtained from the Medline and Indice Medico Espanol (Spanish Medical Index) bases. Characteristics of the study, authors and bibliographic references were analysed. The greatest number of studies were published in Atencion Primaria (81%), making up consistently about 10% of the annual total of articles in this journal. A descriptive design and inferential statistical analysis (comparison of proportions) were mainly used. Average number of authors was 5.14 (C.I. 95%, 4.7 - 5.6). The participation of residents (39%) was greater than that of family doctors and below that of other specialists, suggesting a certain disenchantment with research. The collaboration of different centres (49%) was high, fundamentally Health Centres among themselves and with universities. CONCLUSIONS: PC research in the Community of Valencia appears to be experiencing similar growth to that in the rest of Spain. PMID- 7488682 TI - [Influence of the organizational model on user satisfaction]. AB - OBJECTIVE: To identify in Health Centres (HC), doctors' offices and out-patient clinics the variables among the use, the doctor and organisation, which most affect satisfaction of the Primary Care (PC) user. DESIGN: A descriptive crossover study. SETTING: PC level in three Andalusian cities. PATIENTS AND OTHER PARTICIPANTS: A sample of groups of 460 users of out-patient clinics and doctors' offices and 400 from HCs, all in an urban environment. MEASUREMENTS AND MAIN RESULTS: Three questionnaires were used to gather the independent variables of organisation, doctor and use, and also the dependent variable, or satisfaction, in three dimensions: professional competence, personal qualities and accessibility. The reliability of the scale of satisfaction was analysed using Crombach's alpha; and discriminative capacity, using the Edwards test. The effect of the variables of doctor, user and organisation on HCs and doctors' offices / out-patient clinics was analysed separately with multiple regression. The determination coefficient of the variables for the user at doctors' offices / out patient clinics was higher than the same variables in HCs (p < 0.0001), age being the variable which most affected satisfaction, both in HCs and doctors' offices / out-patient clinics. The determination coefficients for the regressions of the variables of doctor and organisation were higher in HCs than in doctors' offices / outpatient clinics (p < 0.001) in both cases - with the use of the clinical records being the primary variable in both organisations. CONCLUSIONS: It appears that the organisational model of HCs offers more possibilities of increasing user satisfaction by modifying determined features of the doctor and the organisation. PMID- 7488684 TI - [Suitability of a tobacco dependence clinic in primary care]. AB - OBJECTIVE: To evaluate the results of a tobacco dependency clinic after 41 months functioning. DESIGN: A descriptive and retrospective study. SETTING: The tobacco dependency clinic of the Primary Care team at Sant Just Desvern. PARTICIPANTS: 134 smokers who asked for an interview with the nurse in charge of the clinic were attended. MEASUREMENTS AND MAIN RESULTS: 47.01% wanted to stop smoking: 66.42% anticipated difficulties in achieving this. Treatment was counter indicated in 17.16% of the smokers. 57 people (42.54%) stopped smoking of whom 26 then restarted. 35 smokers (26.12%) cut down and 17 (12.69%) changed their attitude. CONCLUSIONS: The clinic fulfilled its initial objective, but we must move towards a strategy of systematic intervention on all smokers within the general clinics. PMID- 7488686 TI - [Rational use of drugs. Viewpoint of the users in the 3d Health Area of Saragossa]. AB - OBJECTIVE: To know the role of drugs and their use from the point of view of the National Health System users. DESIGN: Development of a qualitative method: focal groups of discussion. SITE: Health Area 3 of Zaragoza (Spain) which belongs to the Spanish National Health System. POPULATION: Groups of eight people who are representative of the rural and urban population. MAIN MEASUREMENT AND RESULTS: There were different meeting of one hour and a half for each. All of them started with the same question: What utility, use, and functions have drugs for all of you? All the session were recorded on video-tape and cassette to facilitate its typewriting. The general opinion was that users did not like to take drugs, nevertheless, it was a tool to solve easily and fast a health problem. At the same time, it was a cheap resource comparing to others as massage, health resort, diets... Drugs are seen as interchange currency in the medical bureau. There were critical opinions against abusive consumption of drugs. There is a lack of information concerning the utility and actions of drugs. CONCLUSIONS: The speech of user groups shows opposing points of view related to health professional opinions concerning drugs request from users and the role of drugs in the relationship doctor-patient. PMID- 7488687 TI - [Prevalence study of symptoms and risk factors associated with climacteric]. AB - OBJECTIVES: To describe the prevalence of risk factors for Osteoporosis and Cardiovascular disease and the frequency of appearance of menopausal symptoms; and to study how these are linked to menstrual disorders in premenopausal women. DESIGN: Descriptive study. SETTING: General Medical clinics of an urban PC team in Leganes, Madrid. PATIENTS: 181 women who attended the clinics without appointment for whatever reason over a month. MEASUREMENTS AND MAIN RESULTS: A directed questionnaire on risk factors and 19 symptoms of the menopause was completed. Among the risk factors for Osteoporosis, 52.2% of women consumed less than 500 mg of calcium per day, 8.3% had never given birth and 6.7% presented early menopause. As risk factors for Cardiovascular disease, the high prevalence of arterial Hypertension (32.6%) and Dislipaemia (24.9%) were notable. Only flushes, pain in the joints, migraine, fatigue, nervousness and loss of memory were present in more than 50% of the women. The thirteen remaining symptoms did not reach this proportion. We found no association between classic symptoms of menopause and premenopausal menstrual disorders. CONCLUSIONS: The intake of calcium by women in our health area must be increased. We thought there were reasonable doubts for some classic symptoms of the menopause being accepted as indicators for hormone replacement therapy. PMID- 7488689 TI - [Primary care research: attitude of family physicians trained at the Cordoba Teaching Unit]. AB - OBJECTIVE: To study the opinions and attitudes of a group of Family Doctors concerning various aspects of Primary Health Care (PHC) research. DESIGN: A descriptive observational study. SETTING: Health Centres and Primary Care consulting rooms. PARTICIPANTS: Family doctors trained in the Family and Community Medicine Teaching Unit in Cordoba between 1982 and 1992 (N = 70). There was a 59.7% reply rate. MEASUREMENTS AND RESULTS: Among the obstacles noted for the development of research in PHC, lack of time at work was the main one (82.5%); and as a handicap, lack of understanding how to work out designs was in first place (85.0%); and as a handicap, lack of understanding how to work out designs was in first place (85.0%). 60.0% considered that PHC research is not sufficiently valued in our country and 77.5% thought that it is not taken into account enough on applying for jobs. 78.0% (C.I.95%: 62.0 - 88.9%) took part in a congress giving papers or reports during the last year. 63.4% (C.I.95%: 46.9 - 77.4%) were working on a research project. CONCLUSIONS: In spite of the perceived methodological difficulties and the lack of incentives, the number of family doctors who devote part of their time to research projects is notable and their participation in scientific events is considerable. PMID- 7488688 TI - [Microvascular angina (syndrome X). Presentation of 4 cases]. AB - OBJECTIVE: To revise diagnostic, therapeutic and evolutive specifications of Microvascular Angina. To improve the knowledge about this syndrome among family doctors. DESIGN: Descriptive and transversal study. SETTING: Outpatients belonging to a Primary Care Centre in the metropolitan area of Madrid. PATIENTS AND OTHER PARTICIPANTS: Four patients (2 male and 2 female) aged over 50, diagnosed to have Microvascular Angina. MEASUREMENTS AND MAIN RESULTS: The clinical histories of the patients diagnosed to have Microvascular Angina were used. This syndrome was defined as a typical chest pain starting by an effort, normal electrocardiogram, ischemic ST-segment depression in exercise stress test and angiographically normal coronary arteries. The clinical and electrocardiographic specifications were similar in four cases. A positive exercise stress test and the existence of normal coronary arteries was confirmed in all the patients. All of them were treated with betablokers and/or calcium channel blokers with an effective response in three of them. CONCLUSIONS: It is important to know the existence of Microvascular Angina in Primary Care, first because it is one cause of chest pain and second because it has a good prognosis with few complications in the future. PMID- 7488690 TI - [Effectiveness of serotonergic agonists in the treatment of obese patients]. AB - OBJECTIVE: To compare the effectiveness of two interventions in the treatment of obese patients: hypocaloric diet and hypocaloric diet plus serotoninergic medication. DESIGN: Randomised clinical trial. SETTING: Primary Care. Out-patient Endocrinology clinic. PATIENTS: The trial included 60 obese adults, assigned at random to two groups of 30 each. Both groups were comparable for sex, age, weight and Body Mass Index. Weight controls were performed at 2, 4 and 6 months. RESULTS: There were no significant differences in weight losses at the times studied (2, 4 and 6 months) between the Control (Hypocaloric diet) and Study (Hypocaloric diet plus medication) groups. There were significant weight losses in both groups analysed between 2 and 4 months, with a Body Mass Index which went down from 36.2 to 34.12 and from 35.52 to 33.57 (p < 0.001) in the Control and Study groups, respectively. CONCLUSIONS: No significant differences were observed in the results obtained for obese patients between a hypocaloric diet supplemented by serotoninergic medication and a hypocaloric diet. PMID- 7488691 TI - [Drug-induced gingival hyperplasia]. PMID- 7488692 TI - [Hepatitis B vaccination among school children]. PMID- 7488693 TI - [Blood sugar control and complications of diabetes: the problem of NIDDM]. PMID- 7488694 TI - [Universal screening for colorectal cancer]. PMID- 7488695 TI - [Prevalence of generalized pain of the locomotor system]. PMID- 7488696 TI - Introduction to the Bioelectronic Devices Project in Japan. AB - The Bioelectronic Devices Project was organized in Japan as a 10-year national project and it has been working toward developing fundamental key technologies for designing and assembling innovative information-processing devices by realizing the excellent functions specifically found in molecular assemblies and information processing of living organisms. The project is now in the third year of the second phase (the 8th year of its 10-year duration) and researchers have been trying to elucidate the specific characters of the prototype devices. The outline of this challenging project is given with the latest experimental results. PMID- 7488697 TI - Towards efficient molecular wires and switches: the Brooker ions. AB - In search of materials which may function as molecular wires or switches, analytical models have suggested that the Brooker ions should be particularly interesting. We study them in detail using ab initio, semi-empirical and specially-designed empirical techniques, predicting molecular geometries, charge distributions, and conductivities. Provided molecular symmetry is maintained, odd polyenes and Brooker ions NH2-(CH)+ 2n - 1 - NH2 are shown to conduct significantly better than even polyenes, but the advantage becomes a simple multiplicative factor once solitons form (chains of length ca 20 A). Symmetry lowering is predicted to dramatically decrease the conductivity but introduces the possibility that the Brooker ions may function as molecular switches, having greatly enhanced, switchable, non-linear optical properties. PMID- 7488699 TI - Fundamentals of photoelectric effects in molecular electronic thin film devices: applications to bacteriorhodopsin-based devices. AB - This tutorial lecture focuses on the fundamental mechanistic aspects of light induced charge movements in pigment-containing membranes. The topic is relevant to molecular electronics because many prototypes optoelectronic devices are configured as pigment-containing thin films. We use reconstituted bacteriorhodopsin membranes as an example to illustrate the underlying principle of measurements and data interpretation. Bacteriorhodopsin, a light-driven proton pump, is the only protein component in the purple membrane of Halobacterium halobium. It resembles the visual pigment rhodopsin chemically but performs the function of photosynthesis. Bacteriorhodopsin thus offers an unprecedented opportunity for us to compare the visual photoreceptor and the photosynthetic apparatus from a mechanistic point of view. Bacteriorhodopsin, well known for its exceptional chemical and mechanical stability, is also a popular advanced biomaterial for molecular device construction. The tutorial approaches the subject from two angles. First, the fundamental photoelectric properties are exploited for device construction. Second, basic design principles for photosensors and photon energy converters can be elucidated via 'reverse engineering'. The concept of molecular intelligence and the principle of biomimetic science are discussed. PMID- 7488698 TI - Two-dimensional domain shape transitions in photosensitive monolayer assemblies. AB - Bubble-stripe shape transitions of gaseous domains are observed in Langmuir monolayers of porphyrin/arachidic acid and porphyrin/spiropyran mixtures using fluorescence microscopy. Characteristics of the transition such as critical bubble size are found to be controllable by subphase pH or UV/visible irradiation. These features are discussed in terms of a balance between dipolar repulsions and line tension in the monolayer. PMID- 7488700 TI - Oscillatory phenomena in model membrane: electrical oscillation in lipid impregnated membrane filter induced by alamethicin and controlled by bacteriorhodopsin. AB - A lipid-impregnated membrane filter was able to show reproducibly of an electrical oscillation under constant current stimulation when alamethicin and protamine were present in the chamber whose salt concentration was higher than the other and the membrane was left for about 12 h. In addition, bacteriorhodopsin, which is a light-activated proton pump, was found to control the oscillation. The oscillatory phenomenon was considered to be driven by the alternating change in the ion-selectivity of the membrane between cation and anion. Impedance measurement indicated the existence of lipid rearrangement which might prepare an environment for both alamethicin and protamine to cause the oscillation. PMID- 7488701 TI - 4-Keto-bacteriorhodopsin films as a promising photochromic and electrochromic biological material. AB - Photochromic and electrochromic spectral properties of 4-keto-bacteriorhodopsin (4-keto-BR) embedded in a polymer matrix were studied. The light-induced spectral changes were found to be similar to those for 4-keto-BR in suspension, but the duration of the photocycle is substantially longer (up to ten of h). Application of a constant electric field induces a bathochromic shift of the main absorption band, the amplitude of the field-induced spectral changes, showing a quadratic dependence on the field strength. Polymer films containing bacteriorhodopsin analogs show promise as new spectrally-selective photochromic and electrochromic materials. PMID- 7488703 TI - A method for patterning purple membrane using self-assembled monolayers. AB - A method for patterning purple membrane on a substrate is described. This method involves the use of patterned self-assembled monolayers. The quantity of purple membrane adsorbed to glass terminated by various self-assembled monolayers was measured using angle resolved X-ray photoelectron spectroscopy. It was determined that purple membrane adsorbs to unmodified glass and amine-terminated glass surfaces. Hydrocarbon- and fluorocarbon-terminated glass inhibit the adsorption of purple membrane. To demonstrate a potential patterning methodology, an amine terminated self-assembled monolayer (SAM) was photochemically substituted for a hydrocarbon-terminated SAM using UV radiation. Future work will involve the use of UV radiation in conjunction with a lithographic mask so that specific PM adsorption sites can be defined on a substrate. PMID- 7488702 TI - The cycle of photochromic reactions of a bacteriorhodopsin analog with 4-keto retinal. AB - Photochemical reactions in a bacteriorhodopsin analog with 4-keto-retinal (4-keto BR) were studied by using low-temperature and pulsed laser absorption spectroscopy. A photocycle of the photochemical reactions of 4-keto-BR is proposed, which, unlike the photocycle of native BR, includes several spectrally and kinetically distinguishable M-type and O-type intermediates. PMID- 7488704 TI - Holographic properties of Triton X-100-treated bacteriorhodopsin embedded in gelatin films. AB - Bacteriorhodopsin (b) thin films have been fabricated with varying amounts of the detergent Triton X-100 to measure the effect of this additive on the holographic performance of these thin films. Holographic spectroscopy is used to measure the effect of these detergents on the overall diffraction efficiency as well as on the phase and amplitude components of the overall signal. The diffracted rise and decay kinetics of these materials will also be presented as a function of varying detergent concentration. This research also studied the effect of this additive on the absorptive properties of bR-based thin films. Comparisons of the two complimentary sets of data are drawn. PMID- 7488705 TI - Advances in protein-based three-dimensional optical memories. AB - An oriented bacteriorhodopsin cube is optimized as a potential three-dimensional optical memory medium. Write/read capability is demonstrated by using the photovoltaic signal induced by two-photon absorption. Our results demonstrate that a two-photon induced photovoltage can he detected in a three-dimensional Bacteriorhodopsin (BR) cube as large as 1.6 x 1.6 x 1.6 cm3. The read/write speed, signal to noise ratio, and the laser damage threshold for the protein based three-dimensional optical memory is examined. PMID- 7488706 TI - Riboflavin: a potential material for molecular electronics applications. AB - Riboflavin (Rbf), the simplest manifestation of the isoalloxazine ring, undergoes redox reactions during biochemical processes. The redox reaction can be simulated under electrochemical conditions. The two oxidation states of the isoalloxazine molecule have different optical properties. Rbf embedded in a polyurethane matrix has been shown to undergo colour change on application of an external electric field. Rbf has been modified to form TARbf and TPRbf--the amphiphilic analogues for the convenience of forming ordered two-dimensional organization. The modification has been found to leave native characteristics of Rbf undisturbed. TPRbf forms monolayers at the air-water interface with a collapse pressure of 10 mNm-1. PMID- 7488707 TI - Cross-scale interactions in biomolecular information processing. AB - Multiple modes of processing, drawing on interactions at multiple physical scales, contribute to the information processing power of biological systems. Conformational processing is central, since it affords a close coupling of nuclear and electronic coordinates, and hence a closely interleaved classical nonclassical interface that allows biological systems to draw on the superpositional parallelism of the quantum mechanical wave function. Cells and organisms are pictured as percolation networks that mediate highly selective transductions of information between microscopic and macroscopic forms. PMID- 7488708 TI - Scaling of efficiency in programmable and non-programmable systems. AB - The number of possible interactions available for problem solving is vastly greater for biological systems than for structurally programmable models of the digital type. The difference enables cells and organisms to utilize physical scales and modes of processing that are not accessible to silicon-based technologies, but that could be exploited by biomolecule-based systems. PMID- 7488709 TI - Enzymes, artificial cells and the nature of biological information. AB - Recent progresses in enzyme processing within an expanding array of disciplines make the informational analysis of the whole enzymic complexity of the simplest cellular systems appear as a plausible interdisciplinary enterprise. The term 'artificial cell' could be used as a label to encompass these global information processing models of the cell. In a simplified view, three different degrees of enzymic complexity emerge: self-organization, self-reshaping and self modification. They imply that metabolic enzyme networks endowed with a signaling system and with a 'DNA world', constitute a genuine information processing engine for cellular problem solving, a true molecular 'society'. Philosophically, a new path can be followed to discuss the biological foundations and the paradoxes of the information concept. Other 'informational societies' could be analyzed in a similar way. PMID- 7488710 TI - Dynamical self-organization in biomolecular systems of charge transport. AB - Many non-linear and non-equilibrium aspects of the functioning of biomolecular objects indicate the effects of dynamical self-organization on the level of separate complexes or even individual macromolecules. Charge-conformational interactions provide a natural basis of molecular synergetic phenomena. Considering the charge transfer processes in ionic channels and photosynthetic reaction centres from this point of view, we exemplify recent theoretical and experimental results in its favour. PMID- 7488711 TI - Biological effects of environmental electromagnetic fields: molecular mechanisms. AB - In recent years, epidemiological studies have shown an increased risk of leukemia in children living near electric power distribution lines. Also, medical studies have shown that low frequency electromagnetic (EM) fields accelerate the healing of bone fractures. The stimulation of biosynthesis suggested by both epidemiological and clinical studies is observed in vitro, and indicates that EM fields stimulate the biosynthetic 'stress response' in cells. Studies of EM field effects on enzyme (Na,K-ATPase) function suggest that these effects should be considered in the design of molecular electronic devices. PMID- 7488712 TI - Immobilization of enzymes/coenzymes for molecular electronics applications. AB - A brief survey of immobilization strategies and methods has been presented. Relative merits and demerits are discussed with reference to their end use. PMID- 7488713 TI - A highly stable and selective biosensor using modified nicotinic acetylcholine receptor (nAChR). AB - Methods for developing stable, sensitive and selective bilayer lipid membrane (BLM)-based biosensors are discussed. Stable BLMs were formed over micromachined polyimide apertures. Selective sensors were made by incorporating nicotinic acetylcholine receptors (nAChRs) modified with bispecific antibodies (BsAbs). When two BsAbs, attached to one nAChR, encounter antigen (Ag), channels are blocked. Sensitivity to single Ag molecules would be possible by monitoring closure of individual nAChRs. PMID- 7488714 TI - Optical neural device based on memory-type organic photoconductors. AB - A variable-sensitivity organic-photoconductive device has been developed by using a memory-type organic photoconductor. The device is composed of an input layer and a memory layer acting as a charge generation and a charge transport layer, respectively. A persistent high-sensitivity state, which corresponds to a modulation of synaptic weights in a neuron model, was obtained by blue light illumination. A photochemical reaction of thiomichler's ketone (TMK) in the memory layer was found to be responsible for the memory formation. As a potential application of this memory-type organic photoconductor to an optical neural device, 2-D pattern storage and calculations were performed successfully. PMID- 7488715 TI - Practical approach to implementation of neural nets at the molecular level. AB - Potentialities for implementing simple neural net information processing devices based on chemical and biochemical dynamic media are discussed. This approach gives an opportunity to construct efficient systems capable of performing some primitive operations important for imaging processing. PMID- 7488716 TI - Directional guidance of neurite outgrowth using substrates patterned with biomaterials. AB - The use of geometrically simple networks formed by cultured neurons facilitates the electrophysiological study of biological computation. We used chemically patterned substrates for culturing SK-N-SH human neuroblastoma cells and embryonic rat hippocampal neurons to geometrically control their neurite outgrowth. On patterned substrates (parallel lines, 5-10 microns width), the neuroblastoma cells developed bipolar morphology with long neurite processes (approximately 200 microns) in the presence of retinoic acid. Hippocampal neurons cultured on substrates of hexagonal patterns extended their neurites preferentially along the circumferences of the hexagons and formed geometrically well defined network structures. PMID- 7488717 TI - Molecular quantum computer of neuron. AB - Living cells are controlled by quantum regulators in which the price of action of elementary operations approaches Planck's constant. The description of such systems is based on four principles: (1) minimal price of action principle for control; (2) principle of optimality; (3) minimum irreversibility principle; and (4) the principle of causality. PMID- 7488718 TI - Quantum and biological computation. AB - Ubiquitous internal measurement of material origin and conservation laws, when combined together, uphold biological computation as a specific mode of quantum computation. Internal measurement supplemented by conservation laws can reproduce quantum mechanics or the uncertainty principle in particular. Furthermore, biological computation founded upon internal measurement provides an irreversible enhancement of organization and quantum coherency through non-algorithmic and non programmable procedures of generating variations in accordance with the operation of the uncertainty principle. PMID- 7488719 TI - Inter- and intracellular computation models based on Boolean vs. non-Boolean inconsistency. AB - Systems involving both intracellular and intercellular computation are destined to be described as non-computable. We propose a model for such systems by introducing the inconsistent relation of Boolean and non-Boolean logic. Cellular automata fashioned model exhibits an evolution like class 4 located at the edge of chaos, while there is no local rule for universality existing over the whole space in our model. A system featuring the inconsistent vertical scheme is approximately articulated into a hierarchical system, whose wholeness cannot be deduced by any approximated local rule. In other words, undecidability between a part and whole comprises a hierarchical structure. PMID- 7488720 TI - The information generated in a man-to-man game called "Renju (Go-bang)". AB - A learning experiment was designed using "Renju (Go-bang)". The matches could proceed with prior indefiniteness, distinct from probability, as under the finite VOP (velocity of observation propagation). The information of each situation in the first game SSG and one in the replay SRE were investigated with the basic strategy derived from the "Renju" rules. The behavior of the difference (SRE (i) SSG (i)) suggested that prior indefiniteness turned into definiteness, suggesting that perpetual decision change occurred and that it perpetually allowed observers to construct higher levels of hierarchical learning logic. PMID- 7488721 TI - A programmable interface to neuromolecular computing networks. AB - A programmable interface is provided to a simulated network of reaction-diffusion neurons. The interface allows special 'learn' and 'decide' syntactic constructs to be intermixed with conventional programming constructs. This hybrid combination allows the power of programmability to be combined with the power of adaptability to provide innovative solutions to complex problems. The network uses reaction-diffusion neurons instead of adaline neurons. A mesh topology is used instead of a feedforward topology. The performance of the mesh reaction diffusion network compares favorably with that of conventional feedforward adaline networks. Enhancements to incorporate short- and long-term memory are described. PMID- 7488722 TI - Integrating molecular and digital computing: an information systems design perspective. AB - Biological systems use a non-programmable, but evolvable and efficient mode of information processing that can be traced to its underlying macromolecular basis (Conrad, 1985, Commun. ACM 28, 464-480). Digital computers, on the other hand, are structurally programmable, but not evolvable, nor efficient. In this paper we explore ways to provide effective support of function in organizations by combining the molecular and digital modes of information processing in a synergistic manner. In particular, we look at the potential of molecular computing technology for extending the problem-solving and decision-making capabilities of humans in complex organizational systems. Synergies resulting from the integration of molecular computing (including both human and molecular computing devices) and digital computing, are analyzed from an information systems design perspective. PMID- 7488723 TI - The future of MEBC: panel discussion. AB - The expected developments in the not too distant future (5-10 years) of molecular electronics and biocomputing (MEBC) are discussed. In the short-term, the study of very specific basic phenomena is expected (e.g. conducting polymers, strange electronic states of insulating polymers, bacteriorhodopsin (BR), arrays of molecules, self-organization of biomaterials, very specific biological systems, quantum coherence in cytoskeletal microtubules, optoelectronic information storage, associative memories, pattern recognition, hierarchical nature of biological information). New application fields outside the range of conventional technology (e.g. randomized algorithms, optoelectronic devices, chemical and biosensors, as well as a certain extent of commercialisation) have also been predicted. In the long-term, the study and solution of much deeper (sometimes scientific fiction-like) problems were foreseen, such as the self-organization of biomaterials, artificial self-reproduction, implementation of artificial cell dynamic control structures based on molecular devices for medical and environmental applications and the construction of neuronal computers as aids to the human brain. PMID- 7488724 TI - Proceedings of the International Society for Molecular Electronics and Biocomputing meeting. Gaithersburg, Maryland, September 21-23, 1993. PMID- 7488725 TI - Rapid habituation of auditory responses of locus coeruleus cells in anaesthetized and awake rats. AB - The auditory response of locus coeruleus (LC) neurones evoked by novel tones was investigated in anaesthetized and awake rats. Recording the single unit activity of LC neurones, responses to auditory stimuli are found under anaesthesia as well as in the awake animal. There are three types of LC responses to tone: first and by far the most frequent, a burst of several spikes at onset of the tone; second, a burst at tone offset and lastly, a total inhibition to the tone. All responses present a rapid habituation after the first few presentations of the stimulus. The results support the view that the LC plays a role in mediating responses to environmental changes. PMID- 7488726 TI - Modulation of carbon monoxide production and enhanced spatial learning by tin protoporphyrin. AB - Endogenous carbon monoxide (CO), produced by haem oxygenase (HO), may play a role in hippocampal long-term potentiation (LTP). Its role in learning and memory in intact animals is less well known. Tin protoporphyrin (Sn-PP; 25 mg kg-1, i.p.) effectively but transiently inhibited HO activity in brain homogenates, and improved acquisition in the Morris water maze. Locomotor activity was unaffected, indicating a behavioural specificity of the learning effect. The analogue zinc protoporphyrin (25 mg kg-1, i.p.), which does not pass the blood-brain barrier, did not affect learning. If the observed memory effect is related to inhibition of HO, the role of CO in spatial learning may be different from that suggested by LTP studies. PMID- 7488727 TI - Medullary responses to chemoreceptor activation are inhibited by locus coeruleus and nucleus raphe magnus. AB - The effect of electrical and L-glutamate stimulation of the nucleus locus coeruleus (LC) and nucleus raphe magnus (NRM) on multiunit activity evoked in the nucleus tractus solitarius (NTS) by activation of arterial chemoreceptors (15-25 micrograms kg-1 of sodium cyanoborohydride, i.v.) was studied in rats anaesthetized with urethane (1.1 g kg-1 i.p.). Multiunit discharge of NTS neurones in response to cyanide injection was composed by spikes higher than 200 microV and about 10 Hz frequency. Electrical and L-glutamate stimulation of the LC and the NRM significantly reduced the frequency of the cyanide-induced multiunit discharge. The results show that neurones of the NTS with input from arterial chemoreceptors can be inhibited by LC and and NRM cells, suggesting that these nuclei play a role in controlling chemosensory input at the NTS. PMID- 7488728 TI - Piriform cortex late activity revealed functional spatial heterogeneity. AB - Optical signals were recorded in the in vivo rat piriform cortex (PC) in response to olfactory bulb electrical stimulation. Sometimes the early response was followed by a longer latency component with an occurrence probability of 0.25. In order to compare the early and late activity, the ratio between early (disynaptic) and late wave amplitudes was measured at each recording site on the whole PC. Its spatial distribution revealed that the relative importance of the late activity was larger in the most posterior part of the PC whereas the late wave was rarely observed in the anterior PC. Such a result gave new information on the functional heterogeneity of the PC. PMID- 7488729 TI - [3H]phorbol 12,13-dibutyrate/PKC binding in thoracic spinal cord: no change in amyotrophic lateral sclerosis. AB - [3H]phorbol 12,13-dibutyrate ([3H]PDBu), a selective ligand for various protein kinase C isozymes (PKC), was used to investigate the distribution of [3H]PDBu/PKC binding sites in thoracic spinal cords of patients who died with amyotrophic lateral sclerosis (ALS) and subjects free of neurological disease. In controls, binding of [3H]PDBu was mostly concentrated in the substantia gelatinosa and to a lesser extent in other dorsal horn regions (laminae III and IV). [3H]PDBu binding sites were also present in the ventral horn and laminae X but in somewhat lower quantities. The distribution of [3H]PDBu binding in thoracic spinal cords of patients who died with ALS was unchanged compared to controls and no significant differences were observed in the amount of specific binding in ALS. The present results on the distribution of [3H]PDBu binding sites in human spinal cord are consistent with previous studies of [3H]PDBu binding in rodents indicating that high levels of PKC are present in the dorsal horn. The results also suggest that levels of PKC isozymes are not altered in ALS spinal cord. PMID- 7488730 TI - Carrageenan oedema and spinal Fos-LI neurones are reduced by piroxicam in the rat. AB - Intraplantar carrageenan induced a peripheral oedema and c-Fos like immunoreactivity (Fos-LI) in L4-L5 segments in the dorsal horn (DH) of the rat spinal cord. The total number of carrageenan evoked Fos-LI neurones was 153 +/- 10 per section, with a predominant localization in the superficial and deep laminae of the DH (46 +/- 4% and 32 +/- 2%, respectively). Pre-administered piroxicam (1, 2.5 and 5 mg kg-1, p.o.) dose-dependently reduced the number of Fos LI neurones in both the superficial and deep laminae of DH, with effects being significantly strongest on the deep laminae Fos-LI neurones. The effects of piroxicam on the number of spinal Fos-LI neurones and on ankle oedema were positively correlated. PMID- 7488731 TI - Carbon monoxide as a putative messenger molecule in the feline lower oesophageal sphincter of the cat. AB - The distribution of the carbon monoxide (CO) producing enzymes haem oxygenase (HO) type 1 and 2 were studied in the feline lower oesophageal sphincter (LOS), as were HO activity and functional effects of CO. HO-2 immunoreactivity was observed in nerve cell bodies in the submucosal and myenteric plexus, nerve fibres, non-neuronal cells surrounding smooth muscle bundles, and in arterial endothelium, HO-1 immunoreactivity was confined to non-neuronal cells in the smooth muscle layer. CO production, indicating HO activity, was demonstrated in tissue homogenates. CO relaxed the LOS, and activated the cyclic GMP system. These results show that HO is present in the LOS, and suggest that CO can be generated by neuronal and non-neuronal structures and may have a role as a peripheral messenger. PMID- 7488733 TI - Brain activation during the generation of non-emotional and emotional plans. AB - We studied 12 normal volunteers who were asked to imagine and plan their behavior in emotional and nonemotional situations while their regional cerebral blood flow was measured with positron emission tomography. The dorsolateral prefrontal and posterior temporal cortex were more activated during the nonemotional situation whereas the medial prefrontal cortex and anterior temporal cortex were more activated during the emotional situation. These results demonstrate that distinctive regions of the prefrontal and temporal cortex used to imagine and plan behavior are activated during the expression of emotional and non-emotional plans. PMID- 7488732 TI - Hepatic vagus does not mediate IL-1 alpha induced anorexia. AB - Peripherally infused interleukin-alpha (IL-1 alpha) reduces food intake. Since the innervated liver modulates eating activity via the vagus, we investigated the role of the hepatic vagus in the etiology of IL-1 alpha induced anorexia. Ten male Fischer 344 rats were randomly assigned to hepatic vagotomy (HX-IL-1 group) or sham operation (Sham-IL-1 group), and an internal jugular catheter was inserted in all rats. Another six sham operated rats receiving normal saline i.v. throughout the study period served as general controls. After a 10-day recovery period, HX-IL-1 and Sham-ILI-1 rats were infused with 3 micrograms day-1 of IL-1 alpha for 3 days, followed by a 4 day infusion of saline. During the IL-1 alpha infusion, food intake was reduced at a similar rate and by a similar amount in both vagotomized and sham-operated rats. When IL-1 alpha infusion was stopped, food intake normalized at a similar rate in both HX-ILI-1 and Sham-IL-1 groups. These data indicate that the hepatic vagus is not involved in the etiology of IL 1 alpha induced anorexia. PMID- 7488734 TI - Epidermal growth factor: a potential paracrine and autocrine system within the pituitary. AB - Epidermal growth factor (EGF) secretion and EGF receptor (EGFR) expression were studied by a combination of reverse hemolytic plaque assay (RHPA), which identifies the EGF secreting cells, and immunocytochemical detection of EGFR. Our results showed that 19% of total rat anterior pituitary (AP) cells secrete EGF and 48% of total AP cells possess EGFR. In addition, most of the EGF-secreting cells (73%) possess EGFR. By measuring the size of plaques surrounding EGF secreting cells, we found that EGF secretion per cell is significantly higher in cells that did not express EGFR (1930 +/- 92 microns2) than in EGFR-expressing cells (1551 +/- 89 microns2). Taken together, these results strongly suggest that EGF acts by both an autocrine and paracrine fashion in the rat anterior pituitary gland. PMID- 7488735 TI - Effect of thyrotropin-releasing hormone on microtubules in GH3 cells. AB - The effect of thyrotropin-releasing hormone (TRH) on the tubulin-microtubule system in GH3 cells was investigated. Prolactin (PRL) secretion by GH3 cells was stimulated by TRH at 1, 5, and 10 min of incubation. The polymerized tubulin levels increased from 0.4 units micrograms-1 protein in control cells to 0.6 units micrograms-1 protein in cells incubated with TRH for 1 min. Soluble tubulin levels increased from 0.8 units microgram-1 protein in control cells to 1.2 units micrograms-1 protein in cells exposed to TRH for 5 min. Tubulin levels were similar in control cells and cells treated with TRH for 10 min. These results suggest that TRH-stimulated PRL secretion is accompanied by a shift in the equilibrium between tubulin pools. PMID- 7488736 TI - Cocaine self-administration and naltrindole, a delta-selective opioid antagonist. AB - Recent reports from several laboratories have suggested a role for delta opioid receptors in expressing some of the biochemical and behavioral effects of cocaine. Here, this possibility has been further explored by evaluating the propensity of rats to self-administer i.v. cocaine in the absence or presence of naltrindole, a selective delta opioid antagonist. Following a number of days of stable cocaine intake, and before a day's session, naltrindole (3 or 10 mg kg-1) reduced pressing for cocaine, regardless of the schedule of reinforcement. These data further support the role of processes associated with delta opioid receptors in the ability of cocaine to reinforce its own use. PMID- 7488737 TI - Inhibition of nitric oxide synthase causes anxiolytic-like behaviour in an elevated plus-maze. AB - The action of inhibition of nitric oxide (NO) synthase by NG-nitro-L-arginine methyl ester (L-NAME) (1-20 mg kg-1) on the exploratory behaviour of rats in the elevated plus-maze was studied. L-NAME induced an anxiolytic-like effect in the plus-maze test, showing a reverse U-shape action behaviour, with a maximal effect at 10 mg kg-1. This effect was not related to a non-specific increase in motor activity, since in the open field test L-NAME did not affect locomotor activity of rats. Pretreatment of rats with L-NAME (1-10 mg kg-1) also tended to attenuate the anti-exploratory action of CCK agonist caerulein (5 micrograms kg-1), but this action was not significant. In conclusion, it appears that NO may be involved in the process that can lead to anxiety in the rat. PMID- 7488738 TI - Serotonergic potentiation of photic phase shifts of the circadian activity rhythm. AB - Recent evidence suggests that serotonin may function to regulate the sensitivity of the circadian clock to the resetting effect of environmental light. Here we report that systemic administration of NAN-190, a drug that acts at both postsynaptic and somatodendritic serotonin receptors, potentiates light-induced phase shifts by as much as 250%. The effects of the drug are dose-related and are significant at light intensities between 0.2 and 200 lux. It is proposed that drugs with pharmacological properties similar to NAN-190 may prove useful as chronobiologics to adjust the sensitivity of the circadian system to natural 'zeitgebers'. PMID- 7488739 TI - The CB1 cannabinoid receptor antagonist SR 141716A affects A9 dopamine neuronal activity in the rat. AB - CB1 receptors and their putative natural ligand anandamide, have been tentatively involved in the control of midbrain extrapyramidal function. Electrophysiological activity of dopamine neurones was measured after acute and repeated administration of the CB1 receptor antagonist SR 141716A (0.3-3 mg kg-1) in rats. Acute SR 141716A increased A9, but not A10 cell population response without affecting either their spontaneous firing rate or apomorphine-induced rate inhibition and prevented amphetamine-induced inhibition of A9, but not of A10 cell firing. After repeated administration SR 141716A (1 or 5 mg kg-1) decreased population response of A9 cells, which was reversed by apomorphine. These results suggest that CB1 receptor blockade by SR 141716A interrupts a cannabinoid-like endogenous tone controlling extrapyramidal function. PMID- 7488740 TI - NG-monomethyl-L-arginine, an inhibitor of nitric oxide synthase, increases extracellular GABA in the striatum of the freely moving rat. AB - We investigated the effect of an NO synthase inhibitor, NG-monomethyl-L-arginine (L-NMMA), on the levels of endogenous GABA in the rat striatum using in vivo microdialysis. Rats were perfused with the artificial CSF containing L-NMMA (0.1, 0.3 and 1.0 mM) or its inactive isomer D-NMMA (1.0 mM) for 1 h. Infusion of L NMMA, but not its D-isomer, dose-dependently increased GABA concentration. Co infusion with tetrodotoxin (1 microM) did not antagonize the increase of GABA induced by L-NMMA. These results show that decreased NO activity enhances GABA release even in the absence of depolarization of GABA neurones. We conclude that NO may be directly acting on GABA nerve terminals and tonically inhibiting GABA release or synthesis under basal conditions. PMID- 7488741 TI - Transforming growth factor beta induces a beta-responsive calcium fluxes in neurons. AB - The beta-amyloid (a beta) peptide is a neurotoxic peptide that accumulates in the brains of Alzheimer patients, but is also present in body fluids at subnanomolar levels. The potential effects of these low levels of a beta are unclear. We have recently shown that physiologic levels of a beta increase tyrosine phosphorylation and induce increases in cytosolic calcium. The basement membrane mixture, Matrigel, is required for observation of the a beta-induced calcium response. We now show that transforming growth factor beta (TGF beta) is the active component in Matrigel eliciting the a beta/calcium response. The response to the type of TGF beta varies depending on the cell type with TGF beta 1 eliciting a beta responsiveness in olfactory neuroblasts, and TGF beta 2 eliciting a beta responsiveness in PC12 cells. PMID- 7488742 TI - Antinociceptive defect of beige-J mice reversed by i.c.v. IP3 or myo-inositol. AB - The C57BL/6J-bgJ/bgJ (beige-J) mutation imparts a blunted response to intracerebroventricular (i.c.v.) morphine in the tail-flick test, without altered micro-opioid receptor number or morphine affinity. We now report that co administration of IP3 (36.1 nmol) restored morphine responsiveness of beige-J mice to essentially that of normal littermates (bg+/bg-; ED50 = 3.9 and 3.5 nmol, respectively). IP3 had no effect on morphine-induced antinociception in control animals. Neither myoinositol at 36.1 nmol nor IP6 at the highest testable dose (4.5 nmol) had a similar effect. Myo-inositol at 5.5 mumol restored beige-J responsiveness to that of littermates. These findings implicate some component of the phosphoinositide cycle in the antinociceptive defect of beige-J mice. PMID- 7488743 TI - Calcium influx recruits an additional class of kinases to hyperphosphorylate tau. AB - SH-SY-5Y human neuroblastoma cells were treated with combinations of the kinase inhibitors HA-1004, W-7 and H-7 and calcium ionophore A23187. Microdensitometric analyses revealed that, in the absence of ionophore-mediated calcium influx, PHF 1 levels were reduced by approximately half in cultures treated with HA-1004 or W 7, but were not reduced by H-7. By contrast, the doubling in PHF-1 immunoreactivity that resulted following ionophore treatment was prevented by all three inhibitors. These analyses demonstrate the recruitment of an additional kinase or kinases in tau phosphorylation following calcium influx, and underscore the possibility that de novo hyperactivation of calcium-dependent kinases may be involved in the early events that propagate PHF formation. PMID- 7488744 TI - Induction of p21ras in Alzheimer pathology. AB - Changes in the expression of the small G-protein p21ras, a critical regulator of cell proliferation and differentiation, were investigated by immunohistochemical methods in the temporal cortex of patients with Alzheimer's disease. In both normal brain and in Alzheimer's disease, the H-ras, K-ras and N-ras gene products were equally detected in almost all neurones and occasionally also in glial cells. In Alzheimer's disease an increased expression of p21ras was observed within neuritic plaques as well as in neurones and glial cells closely associated with plaques. Neurones containing tangle-bearing material also showed a high level of expression. It is suggested that activation of p21ras dependent signalling pathways are involved in the processes of neurodegeneration and/or aberrant repair in Alzheimer's disease. PMID- 7488745 TI - Age-related decrease of serotonergic fibres and S-100 beta immunoreactivity in the rat dentate gyrus. AB - The age-related changes of serotonergic fibres and S-100 beta-positive astrocytes in the rat dentate gyrus were examined using immunohistochemistry. A significant reduction in the number of the serotonergic fibres and S-100 beta-positive astrocytes was observed throughout the dentate gyrus of the aged rat. A positive correlation was noted between the decrease of serotonergic fibres and S-100 beta positive astrocytes in the aged rat dentate gyrus. These findings support the hypothesis that the age-related degeneration of serotonergic fibres in the dentate gyrus is caused by the decrease in serotonergic neurotrophic factor S-100 beta in the astrocytes. PMID- 7488747 TI - Glutamate regulation of GDNF gene expression in the striatum and primary striatal astrocytes. AB - The aim of this study was to investigate the regulation of glial cell line derived neurotrophic factor (GDNF) mRNA by activation of glutamate receptors in the rat striatum. We observed an increase in GDNF mRNA levels in the adult rat striatum after administration of subseizure doses of N-methyl-D,L-aspartic acid (NMA) and kainic acid. Since it is unclear whether the upregulation of GDNF occurred in neurons or astrocytes within the striatum, we further investigated whether GDNF gene expression in primary striatal astrocytes in culture could be regulated by glutamate receptor activation. We found that treatment of the cultures with NMA and kainic acid similarly upregulated GDNF gene expression as observed in vivo, suggesting that striatal astrocytes express functional glutamate receptors. Immunocytochemical and nuclease protection analysis revealed that striatal astrocytes expressed the NMDA-R1 subunit. These findings suggest the regulation of GDNF mRNA in the striatum may be mediated by excitation of glutamate receptors via glutamatergic cortical afferents. PMID- 7488748 TI - Expression of the olfactory cyclic nucleotide gated channel (CNG1) in the rat brain. AB - The expression of the olfactory cyclic nucleotide-gated channel (CNG1) was studied in the rat brain. Using RT-PCR, levels of CNG1 mRNA were determined relative to the expression of a constantly expressed gene, alpha-tubulin. RT-PCR showed that CNG1 mRNA was detectable in the pituitary gland, the olfactory bulb, and the cerebellum of adult and 5-day-old rats. A 3.4 kb mRNA was detected in the olfactory bulb by Northern blotting. In situ hybridization analysis showed that CNG1 mRNA expression is present in the olfactory bulb and in the Purkinje cells of the cerebellum. RT-PCR studies on Purkinje cell-enriched cultures obtained from the cerebellum of 16-day-old embryos (E16) confirmed the expression of CNG1 mRNA in these neurones. Our results show that CNG1 is not restricted to the olfactory epithelium but is also present in specific regions of the brain. These results suggest that cyclic nucleotides may act in the regions that possess CNG1 gene expression to affect the electrical activity of certain neurones directly. PMID- 7488749 TI - Potentiation of 5-HT3 receptor-mediated responses by protein kinase C activation. AB - The effect of agents that activate or inhibit protein kinase C (PKC) on the function of recombinant 5-HT3 receptors expressed in Xenopus oocytes was studied. The PKC activator phorbol 12-myristate 13-acetate (PMA) induced a long-lasting increase in the amplitude of 5-HT-activated ion current. The potentiation was maximal at 20 min and had a duration of approximately 60 min. The inactive phorbol ester, 4 alpha-PMA, had no effect on 5-HT3 receptor-mediated current. The PMA-induced potentiation was concentration-dependent over the concentration range 0.1-300 nM. The percentage potentiation by PMA was maximal at low 5-HT concentrations and decreased with increasing concentrations of 5-HT. For current activated by 0.1 microM 5-HT, maximal potentiation (Emax) was 667% of control, the EC50 was 15 nM and the apparent Hill coefficient was 0.99. The PKC inhibitor, staurosporin, antagonized the PMA potentiation; whereas, inhibitors of protein kinase A (PKA) or tyrosine kinase had no effect on this potentiation. The observations show that PMA can potentiate 5-HT3 receptor-mediated responses and suggest that this potentiation is mediated by activation of PKC. PMID- 7488750 TI - Single-stage surgery for rehabilitation of the edentulous mandible: preliminary results. AB - Rehabilitation of the edentulous mandible by implant-supported bridgework is generally carried out as a two-stage surgical procedure, followed 4 to 5 months later by fabrication and attachment of the prosthesis. Use of a single-stage surgery with controlled immediate loading of the nonsubmerged implants can significantly simplify the logistics of treatment and provide cost/benefit advantages. The results of this study, up to a 2-year follow-up period, suggest that the clinical performance of the procedure compares favorably with the traditional two-stage method. The learning objective of this paper is to present the evaluation results of single-stage surgery with controlled immediate loading. PMID- 7488752 TI - Implantology. PMID- 7488746 TI - Differential effect of functional olfactory deprivation on synaptic vesicle proteins in rat olfactory bulb. AB - The effects of functional odour deprivation on three different proteins associated with the membrane of the synaptic vesicle were examined in the rat olfactory bulb. Six weeks after neonatal unilateral nostril closure, Rab3a, a ras like GTPase, was down-regulated in the odour-deprived bulb in the same manner as tyrosine hydroxylase. In contrast, synaptophysin, a protein of the channel family, and SV2, a putative transporter protein, were not altered. These results suggest that afferent activity is a factor controlling the level of some, but not all, proteins associated with presynaptic vesicles. PMID- 7488751 TI - Mastication forces and implant-bearing surface. AB - This article discusses the shift of emphasis in implantology from the phenomenon of osseointegration of implants to the accurate fabrication of the prosthesis, recognizing the implant/prosthesis unit as an actual replica rather than a replacement of the missing tooth. The study of kinetics with its succession of mandibular cycles is used to discuss the integration of mastication and occlusion. The importance of periimplant ligament in natural teeth is discussed, along with the impact of its absence on implant mobility. The impact of the bone types and the root surface architecture on the implant-bearing surface is presented along with the stress of the mastication forces. All these factors have to be considered in the treatment planning and effectively communicated to the laboratory. The learning objective of this article is to provide updated information in those areas for the reader. PMID- 7488753 TI - Changes in dental practice. PMID- 7488754 TI - Criteria for implant selection: surgical and prosthetic considerations. AB - With the availability of osseointegrated implants in an assortment of diameters as well as lengths, the implant selection process has become more complex. To expand treatment options and optimize outcome, implant selection should be guided by a number of surgical considerations and prosthetic requirements specific to each implant site. Small diameter implants are appropriate in areas of narrow ridge dimension or where prosthetic space is limited. Wide diameter implants are well suited to posterior regions with poorer bone quality and restricted vertical height. They provide a higher degree of implant-to-bone contact, increased stability, reduced screw stress, and an improved emergence profile. The learning objective of this article is to discuss these considerations and to outline the implant selection criteria. PMID- 7488755 TI - Prosthetically formulated natural aesthetics in implant prostheses. AB - The discrepancy in the anatomic dimensions between an endosteal root-form dental implant and the root of a natural tooth can compromise the aesthetic appearance of the final restoration. For the past five years the authors have used an abutment system, the components of which are designed to enable the clinician to achieve a predictable aesthetic result. This article describes and illustrates the anatomic abutment system (AAS). Until recently, the restoration usually began at the gingival line where a cylindrical implant emerged to be restored with a noncylindrical prosthesis. The new system uses components which compensate for the difference between the diameter of the natural tooth root and the endosteal implant. The importance of teamwork is emphasized, and a step-by-step procedure is presented, indicating which steps are performed by the combined team and which are the responsibility of the laboratory or the restorative dentist. The learning objective of this article is to introduce this system to the reader. PMID- 7488756 TI - Chin bone harvesting for autogenous grafting in the maxillary sinus: a clinical report. AB - The sinus lift has become a common dental procedure when insufficient bone exists to support endosseous dental implants. The goal of the procedure is to augment existing bone in the posterior maxilla. Various resorbable and nonresorbable allograft, xenograft, and synthetic materials and mixtures, with and without autogenous bone, have been used for sinus augmentation. The authors prefer a mixture of autogenous bone with resorbable bovine xenograft, utilizing as much autogenous bone as possible. The chin offers an ideal site for bone harvesting because of its advantages over other potential sites, such as iliac crest, rib, and calvarium. These advantages include ease of accessibility, relatively simple office-based surgery, minimal morbidity, and the benefits of utilizing intramembranous versus endochondral bone. This article describes the harvesting and the sinus lift procedures. The learning objective of this article is to present this particular procedure using a clinical report to illustrate. PMID- 7488757 TI - Seasonal variations in circadian rhythms persist in gonadectomized golden-mantled ground squirrels. AB - Circadian wheel-running activity was monitored in intact and gonadectomized female and male golden-mantled ground squirrels maintained in a 14L:10D photoperiod at 21 degrees C. Marked advances in phase angle of activity onset and increases in the duration of the daily active phase occurred each year in intact animals coincident with the phase of reproductive activity. Surprisingly, long term gonadectomized squirrels of both sexes manifested similar seasonal changes in the circadian system. Seasonal circadian variations in locomotor activity are generated by a circannual clock independent from and not influenced by concurrent changes in secretion of gonadal hormones. PMID- 7488758 TI - Twelve-hour phase shifts of hamster circadian rhythms elicited by voluntary wheel running. AB - Running in a novel wheel can serve as a nonphotic zeitgeber to entrain or phase shift circadian rhythms in hamsters. In this study, hamsters were entrained to a light:dark schedule of 14:10 h but had no access to running wheels. At four different phase points of the light cycle, hamsters were transferred to constant darkness and provided with running wheels. All hamsters began running shortly after transfer and were allowed to continue running at their own volition. Approximately 20% of the hamsters transferred at zeitgeber time (ZT) 23 (ZT 12 = lights out) ran more than 4 h after transfer and showed phase advances of the circadian activity rhythm by as much as 15 h, while hamsters that ran less than 4 h on average did not phase shift. A similar result was observed for hamsters transferred at ZT 2. Hamsters transferred at ZT 5 and 8 also did not phase shift if they ran less than 4 h, although the relation between longer runs and phase shifts became less evident. A sustained run in excess of 4 h appeared to be associated with large phase advances. These results show that under certain conditions, a single sustained bout of wheel-running activity is capable of phase shifting the circadian pacemaker by more than 12 h. PMID- 7488759 TI - Diel pattern of locomotor activity in populations of root voles, Microtus oeconomus. AB - Surface locomotor activity of root voles (Microtus oeconomus) during summer and autumn was recorded with passage-counters in four experimental populations in 0.5 ha outdoor enclosures. Populations differed considerably with respect to density, geographical origin and social behavior of animals, and year of observation. The general characteristics of diel activity patterns, however, were very similar among populations. Root voles were generally more active during daytime than during the night, and high activity always occurred around sunrise and sunset. The most prominent feature of the activity patterns was a clear-cut ultradian rhythm at both day and night, resulting in seven activity bouts per 24 h. Pattern adjustment to changing daylength was achieved by an interaction between endogenous ultradian rhythmicity and resetting of the ultradian rhythm at both sunrise and sunset. The daytime rhythm had a period length of 3.0 to 3.5 h, while the nighttime rhythm had a significantly longer period length of 3.5 to 4.0 h. Data in the literature suggest that this mechanism of triggering activity may also occur in other microtine species. PMID- 7488762 TI - Circadian rhythm of body temperature in an ectotherm (Iguana iguana). AB - Ectothermic animals regulate their body temperatures primarily by behavioral adjustment in relation to the thermal characteristics of the environment. Several studies have shown that some vertebrate ectotherms may show a daily pattern of body temperature selection when given a choice of environmental temperature. The pattern of body temperature selection free-runs when the animals are kept in constant darkness, demonstrating the existence of circadian regulation. To test whether there might also be a low amplitude circadian rhythm of body temperature itself, we examined the pattern of body temperature and locomotor activity of the lizard Iguana iguana held in a constant environmental temperature. Both variables were recorded for 3 days in a light:dark cycle and then for 10 days in constant dim light (0.1 lux). Under these conditions the body temperature of the lizard oscillates with a circadian period as does the locomotor behavior. These results demonstrate for the first time that ectothermic animals may display physiologically generated circadian rhythms of body temperature similar to those recorded in endotherms. In some animals the circadian rhythms of body temperature and locomotor activity showed different free-running periods, demonstrating that the body temperature rhythm was not caused by locomotor activity and suggesting internal desyncronization of the two rhythms. PMID- 7488761 TI - The circadian rhythm of thermoregulation in Japanese quail. II. Multioscillator control. AB - Most biochemical, physiological, and behavioral processes in vertebrates show significant daily rhythms. Under constant conditions, these rhythms exhibit an endogenous periodicity around 24 h showing that they are driven by an internal circadian clock. In Japanese quail, the circadian clock driving activity and body temperature rhythms is functionally organized as a dual-oscillator system. Under certain conditions, such as switching birds from light:dark (LD) 12:12 to continuous darkness (DD), the body temperature rhythm splits into two circadian components that free-run independently before recoupling in a normal phase relationship. The behavior of the activity rhythm parallels that of the body temperature rhythm, supporting the hypothesis that both rhythms are driven by the same set of oscillators. In some instances, recoupling fails to occur and birds continue to exhibit two circadian components that free-run independently. Dual oscillator control of body temperature was observed in normal birds, pinealectomized birds, and optic nerve sectioned birds. However, birds were rendered arrhythmic by complete eye removal. It is proposed that the central circadian system (suprachiasmatic nuclei?) acts as a complex pacemaker that is functionally organized as two sets of oscillators and that circadian input from the eyes is necessary to preserve the integrity of this complex pacemaker. PMID- 7488760 TI - Synchronization of circadian rhythms of house sparrows by oral melatonin: effects of changing period. AB - House sparrows (Passer domesticus) whose circadian rhythms of locomotor activity and feeding had been abolished by pinealectomy were held in constant dim light and periodically exposed to melatonin in the drinking water. By alternating 8 h of melatonin water with variable phases of tap water, rhythms with periods (T) ranging from 21 to 27 h were produced. When melatonin was administered in rhythms with periods of 23, 24, and 25 h, feeding and locomotion behavior of most birds were rhythmic and synchronized with the exogenous melatonin rhythm. The rest phase coincided approximately with the phase of melatonin availability. Under melatonin cycles < 23 h and > 25 h, fewer birds had synchronized rhythms. Nonsynchronized birds were either arrhythmic or they expressed free-running rhythms. Under melatonin rhythms with periods between 23 and 26 h, the phase angle difference between defined phases of the behavioral rhythms and the melatonin rhythm became more positive with increasing T. These data are consistent with the hypothesis (a) that periodic exogenous melatonin can substitute, at least to a certain degree, for the endogenous plasma melatonin rhythm normally resulting from the periodic melatonin secretion by the pineal gland, and (b) that this melatonin rhythm acts on another oscillator, possibly the SCN, as part of the overall circadian pacemaking system. PMID- 7488763 TI - Circadian rhythms of feeding activity in sea bass, Dicentrarchus labrax L.: dual phasing capacity of diel demand-feeding pattern. AB - The nocturnal versus diurnal feeding patterns of sea bass under controlled experimental conditions were studied in order to investigate the existence of such a dualistic feeding behavior. The animals (six groups of 4 animals and 8 single fish) were held in tanks filled with recirculating salt water and installed in a "chronolab" under constant conditions (23.5 degrees C and 2.4% salinity). The fish were given access to self-demand feeders and first exposed to a photoperiod regime of 12:12 (12 h light, 70 lx, and 12 h dark, complete darkness) and then to light:dark (LD) pulses (40 min light, 40 min dark). The LD 12:12 cycle was reversed by doubling the light period in day 16, and reversed again in day 27 by doubling the dark period. The circadian rhythm of food demand was strongly synchronized with the LD cycle, and fish exhibited both diurnal and nocturnal patterns. In most fish, the shift of the feeding rhythm to the new LD cycle was very fast for each reversal (bringing forward or delaying their feeds 12 h), indicating a weak participation of an endogenous circadian rhythm. However, when submitted to LD pulses, fish began to free-run with a periodicity of about 23 h and kept feeding in the light or dark phase according to their prior behavior. The existence of a dualism in the diel feeding pattern in sea bass was thus clearly demonstrated and it appeared that the diurnal and nocturnal behavior did not depend exclusively on a circadian phase inversion of the feeding rhythms as this pattern of behavior was enhanced under ultradian LD pulses. The ecological implications of such dual capacity and the switch from one type of phasing to another are hitherto unknown and need further field and laboratory investigation. PMID- 7488764 TI - The effects of period mutations and light on the activity rhythms of Drosophila melanogaster. AB - Strains of Drosophila melanogaster homozygous for alleles of the period gene (perO, perL, perS, and per+) were reared for multiple generations either in light:dark cycles (LD), continuous illumination (LL), or chronic darkness (DD). The locomotor activity of adult flies from these cultures was monitored in either LL or DD. Flies that were reared and tested in DD had a lower proportion of individuals with normal circadian rhythms than flies reared in LD or LL and tested in DD. The activity rhythms of DD-reared DD-tested animals, when present, showed phase coherence within two out of seven populations, while 8 out of 10 LL reared DD-tested showed phase coherence. Flies tested in LL were largely devoid of circadian rhythms regardless of their rearing environment. Ultradian rhythms were more evident under conditions disruptive to circadian rhythmicity, but were observed in the presence and absence of circadian rhythms. The periods of the ultradian rhythms of LL-reared DD-tested and LD-reared DD-tested flies varied significantly among genotypes, while in other rearing and testing regimes, no relationship was found. PMID- 7488766 TI - The effects of congenital heart disease and cardiac surgery on liver blood flow in children. AB - Children with congenital heart disease may have some degree of hepatic impairment, with further impairment developing shortly after surgical correction of the cardiac defect. The redistribution of organ blood flow that occurs during cardiopulmonary bypass implicates ischaemia as one of the principal causes of injury. The aim of this study was to measure liver blood flow in children with congenital heart disease and to determine both the effects of cardiopulmonary bypass and the consequences of corrective surgery. Indocyanine green clearance and auricular densitometry, were used in 31 children. In 83% we demonstrated a reduced liver blood flow, with a mean percentage disappearance rate (PDR) of 12.9% (SEM +/- 1.2). This finding was unrelated to the patient's age, the type of congenital heart defect or the presence or absence of cyanosis. During cardiopulmonary bypass, hepatic perfusion was further reduced in 77% of children, by an average of 67%, out of proportion with the iatrogenic reduction in total body flow. Six hours after surgery, liver blood flow had increased significantly above preoperative levels (p < 0.001; t-test) to approximately normal values with a mean PDR of 20.4% (SEM +/- 1.5). PMID- 7488765 TI - The validation of auricular densitometry for indocyanine green clearance measurement of hepatic blood flow during and after cardiopulmonary bypass in children. AB - The clinical measurement of hepatic perfusion is complicated by a dual blood supply and the invasive nature of the majority of techniques available. The aim of this study was to validate indocyanine green clearance and noninvasive auricular densitometry as a measure of hepatic perfusion in the context of paediatric cardiac surgery. The effects of different dye concentrations on densitometer recording were assessed in vitro and found to have a linear relationship. Similarly, variations in haematocrit, within the range 21-47%, also had little effect on accuracy. Comparison of densitometry and direct blood sampling with plasma spectrophotometry in six postoperative, normothermic children showed no significant difference between the noninvasive and invasive techniques (r = 0.968; p > 0.05, t-test). Comparison in 10 hypothermic children during cardiopulmonary bypass also showed no significant difference between the two methods, provided that no further cooling or rewarming took place (r = 0.83; p > 0.05, Wilcoxon test). Noninvasive auricular densitometry can, therefore, provide a reliable assessment of hepatic perfusion in children undergoing cardiac surgery. PMID- 7488767 TI - Endotoxaemia detected during cardiopulmonary bypass with a modified Limulus amoebocyte lysate assay. AB - Cardiopulmonary bypass (CPB) is associated with blood heparin level fluctuations and a reduction in haematocrit due to crystalloid haemodilution. The effect of these changes on the reliability of the Limulus amoebocyte lysate (LAL) chromogenic microassay for the measurement of plasma endotoxin was assessed in vitro. It was shown that the assay could be significantly compromised by twofold haemodilution which can occur during CPB. The interference effect on the assay caused by CPB-associated heparin was not significant if a comparatively large amount of heparin (25 IU/ml) was added to the blood at the time of sampling. The effect of haemodilution was counteracted by prediluting plasma samples with crystalloid by a factor dependent on the sample haematocrit (to ensure that the proportion of plasma was similar in all samples). A correction was then required to determine the endotoxin level in the original sample. The modified assay was used to determine sequential plasma endotoxin levels in 14 patients undergoing hypothermic nonpulsatile CPB. Endotoxaemia occurred at the time of aortic cross clamp release and reached a peak of 48.9 +/- 12.9 ng/l shortly before the end of CPB, which was significantly higher than baseline values pre-CPB (p < 0.05). Thereafter, there was a decline in endotoxin levels to 28.9 +/- 13.6 ng/l 24 hours later which was still significantly higher than baseline levels (p < 0.05). Peak endotoxaemia was a predictor of protracted hospital stay when compared with haemodynamic and tissue perfusion parameters. PMID- 7488768 TI - Influence of different flow modi during extracorporeal circulation on endothelial derived vasoactive substances. AB - Influences of shear stress on endothelin (ET) as well as prostacyclin (PGF) levels are common findings in different experimental settings. Thus, plasma levels of both substances seem to be a good tool to verify if different flow modi can be produced in blood vessels by generating pulsatile flow with a roller pump. In the present study, 20 patients scheduled for elective aortocoronary bypass operation were divided into two groups at random. One group was perfused with nonpulsatile (CON-group) and the other with pulsatile flow (PULS-group) during extracorporeal circulation. ET and PGF plasma levels were monitored perioperatively together with parameters of renal function and haemodynamic data. ET values were only slightly elevated at the end of extracorporeal circulation (mean baseline value; CON 3.1 pg/ml and PULS 3.2 pg/ml; mean maximal values at the end of cardiopulmonary bypass (CPB) 4.0 pg/ml and 3.9 pg/ml respectively). Prostacylin values (median baseline values: CON 56.7 pg/ml and PULS 57.1 pg/ml) peaked at the end of operation (median CON 117.8 pg/ml and PULS 137.5 pg/ml respectively) with a subsequent small decrease. No differences between the groups could be observed at any time point with respect to vasoactive substances, urine output (CON 6.5 ml/min and PULS 6.2 ml/min) or haemodynamics during CPB. This confirms studies emphasizing that no effective microvascular pulsatile flow is generated by conventional pulsatile flow-generating devices. In the present study, normothermia and a constant flow rate were maintained during CPB. Aortic cannulae (body-surface-related not maximal large diameters) were inserted. Altering these procedures may have led to more pronounced differences between the groups. All patients had an uneventful course after the operation. Similar to other reports, the present study was not able to demonstrate any benefit of pulsatile perfusion during extracorporeal circulation. PMID- 7488769 TI - Continuous retrograde cerebral perfusion supplies substrates for brain metabolism during hypothermic circulatory arrest. AB - Ten patients underwent replacement of ascending aorta and/or aortic arch with aneurysm or dissection, using hypothermic circulatory arrest (HCA) with retrograde cerebral perfusion (RCP). RCP was administered through the superior vena cava cannula continuously during HCA (15 degrees C to 20 degrees C). Mean HCA time was 32 minutes (range, 18-45 minutes). To assess the metabolic changes during RCP, blood samples were taken from carotid arteries and the superior vena cava cannula simultaneously, five minutes after the onset and five minutes prior to termination of continuous retrograde cerebral perfusion (CRCP) for analysis of blood gas and glucose level. One patient died intraoperatively due to left ventricular failure. Nine patients survived their operations and all except one with stroke due to partial intimal flap obstruction of innominate artery awoke neurologically intact within four to six hours. One patient died on the postoperative fifth day due to septic shock following resection of ischaemic bowel due to dissection involving the mesenteric artery. Oxygen saturation, pH and glucose level were all found to be lower in blood back-bleeding from the carotid arteries than in blood perfused through the superior vena cava cannula at all sampling times during HCA and CRCP (p < 0.05). Although oxygen and glucose extraction is not only from brain tissue, these data demonstrate the efficacy of CRCP in supplying substrates for brain protection. CRCP is a reliable method as an adjunct to HCA for brain protection. PMID- 7488770 TI - Regional cerebral oxygenation during cardiopulmonary bypass. AB - A significant number of patients suffer transient neuropsychological dysfunction after coronary artery bypass graft (CABG) surgery. Recent studies have implicated reduced levels of O2 supply/demand ratio during the rewarming phase of cardiopulmonary bypass (CPB). Using a near infrared spectroscopy (NIRS) system that permits continuous noninvasive monitoring of regional cerebral O2 saturation (rSO2), we investigated rSO2 during CPB. Following Institution Review Board approval and informed consent, 10 patients undergoing elective CABG were studied. Nonpulsatile CPB with a membrane oxygenator, haemodilution and alpha-state pH management was used. The NIRS system consisting of two low-power laser diode sources (780 nm and 810 nm) with a photodetector placed on the frontal cortex was used to measure continuously haemoglobin, oxyhaemoglobin and cerebral blood volume. Continuous rSO2 measurements were obtained before, during moderate hypothermia, and during the rewarming phase of CPB. A rSO2 < 50% was defined as abnormal. Onset of rSO2 < 50% was at 32 degrees C, and the highest incidence occurred during the late stages of rewarming (35-37 degrees C). On rewarming from hypothermic CPB, 70% of our patients sustained a rSO2 < 50% for an average total duration of 9.3 minutes, primarily at the end of the rewarming period. This suggests a transient global imbalance in cerebral O2 supply and demand in a large percentage of patients during the rewarming phase of CPB. However, it has not been determined how long a rSO2 < 50% can be permitted before neuropsychological dysfunction is induced. Additional studies correlating psychometric testing with rSO2 measurements should prove useful in detecting episodes of impaired cerebral oxygenation, and help define rewarming protocols. PMID- 7488771 TI - Comparison of the effects of centrifugal versus roller pump on the immune response in open-heart surgery. AB - We compared the effects of a centrifugal pump with those of a roller pump on immune responses in 26 coronary artery bypass surgery patients during cardiopulmonary bypass (CPB). The patients were randomly allocated into a (Biomedicus) centrifugal pump group and a (Stockert) twin roller pump group. Leucocyte and differential counts; percentages of lymphocyte subpopulations (CD3 , CD4-, CD8-, CD16-, CD20- and CD25-positive lymphocytes) and monocytes (CD14); phytohaemagglutinin-, concanavalin A-, and pokeweed mitogen-induced and unstimulated proliferation of separated lymphocytes; unstimulated and pokeweed mitogen-stimulated production of IgG, IgM, or IgA; and plasma fibronectin, C reactive protein and serum albumin concentrations were measured preoperatively, immediately before CPB, immediately before aortic declamping and on the first postoperative morning. Significant changes were seen in these variables, but no differences occurred between the groups. PMID- 7488772 TI - In vivo testing of a pulsatile implantable impeller pump as a left ventricular assist device used in calves. AB - A pulsatile implantable impeller pump was tested as a left ventricular assist device in five calves. The experiments lasted for 4-11 days. Death or termination was mainly due to respiratory complications or bleeding, irrelevant to the pump itself. As indicators of haemolysis, thrombogenesis, renal and hepatic functions, free haemoglobin (FHb), haematocrit (Hct), platelet number (Plt), lactate dehydrogenase (LDH), blood urea nitrogen (BUN), creatinine, serum glutamic oxalacetic transaminase (GOT) and total bilirubin were measured preoperatively, at the beginning of the pumping (pump on), six hours later and every day thereafter. The data indicated that the pump caused no severe blood damage or organ dysfunction. Thus, the feasibility of a pulsatile centrifugal pump was demonstrated. The pump with its driver weighs 110 g and is capable of delivering a blood flow up to 8 l/min against 100 mmHg mean pressure. PMID- 7488773 TI - Coronary artery bypass grafting in a patient with haemophilia B. AB - Patients with coagulation disorders present the entire open-heart surgical team with an increased challenge. A patient with a known history of moderately severe Factor IX deficiency (2.4% activity) was evaluated for coronary artery disease. Cardiac catheterization revealed a 99% right coronary artery lesion, a long 99% circumflex lesion and normal left ventricular function. Sextuple coronary artery bypass grafting was performed with the aid of aprotinin and Factor IX transfusions. The patient's platelet count after cardiopulmonary bypass was 65,000/mm3, down from a preoperative level of 172,000/mm3, requiring the transfusion of six units of pooled platelets immediately postoperation. The patient was extubated five and a half hours after arriving in the Intensive Care Unit, and his chest-tube drainage after the first 24 hours was 373 ml. Other than a transient episode of atrial fibrillation on the third postoperative day, the patient had an uneventful postoperative course and was discharged on the sixth postoperative day. With the use of aproptinin and the newer monoclonal antibody purified Factor IX concentrates that have been developed, many of the added risks of performing open-heart surgery on patients with haemophilia B are greatly reduced if not eliminated. PMID- 7488774 TI - Minimally invasive coronary artery bypass grafting: a new method using an anterior mediastinotomy. AB - The benefit of internal mammary artery (IMA) grafting as a long-lasting intervention for coronary artery disease is well recognized. However, largely because they are less invasive, catheter based alternatives are frequently chosen, particularly to treat single or double vessel disease. To retain the advantages of the IMA graft, and to offset the invasiveness of conventional coronary artery bypass grafting, we developed a new minimally invasive method using an anterior mediastinotomy for treating left anterior descending (LAD) or right coronary artery disease, or both. Feasibility studies using 16 pigs and a human cadaver led to approval by the Institutional Review Board for use of this procedure to treat six patients (four men, two women; mean age, 63.8 +/- 13.6 [SD] yrs) who granted informed consent. Pedicle dissection of the IMA, using video assisted thoracoscopy if necessary, was made through a 2- to 3-inch horizontal anterior mediastinotomy. The underlying LAD artery was grafted during femoral vessel cardiopulmonary bypass, with cooling to 30 degrees C, induced ventricular fibrillation, and left ventricular venting if required. Transesophageal echocardiography performed after bypass showed that two patients maintained normal wall motion and four had improvement from the original impairment. One patient suffered a recurrence of angina 4 weeks after the procedure; recatheterization showed an acutely angled IMA, subsequently corrected by balloon angioplasty. The results of follow-up dobutamine echocardiographic stress tests were negative in all patients. With this minimally invasive approach, the procedure should provide the benefits of IMA grafting with shorter hospital stay, more rapid recovery, and less overall cost. PMID- 7488775 TI - The technique of radial artery bypass grafting and early clinical results. AB - There is renewed interest in the use of the radial artery as a conduit for coronary artery bypass. Fifty patients underwent bypass surgery using the radial artery in addition to other conduits between November 24, 1992 and November 8, 1994 at our institution. The mean age was 54.4 +/- 9.1 years (mean +/- SD) and 47 of the patients were male. There were 3.6 +/- 0.9 and stomses per patient, of which 2.2 +/- 0.4 were arterial anastomoses. The most common target vessel for the radial artery has been the obtuse marginal (58.8%), with the aorta as the usual site for proximal anastomosis (80.4%). There have been no ischemic hand complications and no radial nerve deficits. There have been no early or late deaths and no myocardial ischemic complications related to the use of the radial artery. We present the techniques used at our institution for the use of the radial artery as a conduit for coronary artery bypass. PMID- 7488776 TI - Comparison of fresh and glutaraldehyde-treated autologous stented pericardium as pulmonary valve replacement. AB - The use of fresh autologous pericardium in valve surgery has shown poor results in the past mainly due to thickening and retraction. Recently, it has been suggested that a short treatment with glutaraldehyde might radically change its behavior. In an attempt to determine whether this disparity in results is due to the glutaraldehyde treatment or to a better present-day surgical technique, fresh and glutaraldehyde-treated autologous pericardium was mounted in a frame and implanted in the pulmonary position of adult sheep. Six survivors obtained in each group were sacrificed between 2 and 8 months in the "fresh" group and between 2 and 6 months in the "glutaraldehyde-treated" group. Macroscopically, the fresh pericardium became thickened and retracted in all specimens, eventually resulting in severe regurgitation, while the glutaraldehyde-treated, although slightly thickened, retained its pliability without significant retraction. Microscopically, viability of the central core of the collagen was more often preserved in the fresh pericardium. Endothelialization was irregular. In conclusion, short glutaraldehyde treatment seems to improve the results of autologous pericardium mounted on a valve stent. Its effect on calcification remains to be ascertained. PMID- 7488777 TI - Clinical experience with a total artificial heart as a bridge for transplantation: the pitie experience. AB - Since April 1986, 82 patients have received a pneumatic total artificial heart, 62 a JARVIK-7, and 20 a Cardiowest. The duration of support ranged from less than 1 day to 603 days (mean duration: 27 +/- 82). The indications were for acute shock (38 cases) or for chronic deterioration on the transplant waiting list (44 cases). The etiology was mainly due to idiopathic and ischemic cardiomyopathy. With the help of our scoring system, we divided our patients in three groups: Chronic Implantation, represented by two females staying on device for 6 and 19 months, respectively; a High-Risk group of 29 patients characterized by high-risk indications; graft failure, rejection, postcardiotomy patient, postpartum cardiomyopathy, and valvular and congenital reoperation. In addition, the dilated and ischemic cardiomyopathy patients with a score over 6 were included in this group; and an Elective Indication group (51 patients) represented all of the dilated and ischemic cardiomyopathy patients with a score under 6. Due to the shortage of donors, our criteria for transplantation are very strict. Transplants should be made only in cases of hemodynamic stability, on an extubated patient with normal renal and liver functions, without coagulation problems or infection. With such criteria, in the high-risk group, only four patients could be transplanted and of these two are still alive. In contrast, in the elective group, 31 were transplanted (61%), and 71% of these patients were discharged. The rate is improved in the most recent cases, with 90% of the Cardiowest patients being survivors. PMID- 7488778 TI - Myocardial revascularization using branches of the left internal mammary artery. AB - To increase the number of arterial grafts in patients with multivessel coronary disease, we have used branches of the left internal mammary artery (LIMA) as bypass grafts. From March 1990 to June 1993, 15 patients (13 males and 2 females) aged 38 to 65 years (mean 57 +/- 8), received a total of 30 LIMA branch anastomoses. The two terminal branches were used to perform a "Y" graft to the left anterior descending coronary artery territory in 13 patients. A pericardiophrenic branch was used in four cases. Five patients (33.3%) had complete myocardial revascularization using IMA only. There were neither early nor late deaths and all patients are free from angina at a mean follow-up of 20 months (6 to 45 months). However, early postoperative cardiac catheterization, performed in 12 patients (80%), revealed a 20% occlusion rate for each terminal branch and a 50% occlusion rate for the pericardiophrenic branch. Technically challenging due to the small diameter of the grafts, the terminal LIMA branches should be limited to selected patients when more conventional arterial or vein conduits are not available. More proximal branches have a higher occlusion rate and their use is not recommended. PMID- 7488779 TI - Closure of isolated ventricular septal defect with detachment of the tricuspid valve. AB - Detachment of the septal leaflet of the tricuspid valve is an alternative technique for obtaining complete visualization of a perimembranous ventricular septal defect (VSD) in cases where the VSD is obscured by the chordae tendineae or a pouch formation of the septal leaflet. This method presents theoretical concerns because it has the potential for causing postoperative valvular insufficiency. We therefore evaluated valvular function in patients who underwent VSD closure with detachment of the tricuspid valve. In a consecutive series of 153 patients who underwent VSD closure using a transatrial approach, 13 had incision of the tricuspid valve. Follow-up echocardiographic studies were performed on these patients at least 1 year following operation. There were no operative deaths. Color Doppler echocardiography revealed no residual shunt in any of these patients. Ten patients had no evidence of tricuspid stenosis or regurgitation. One patient had trivial tricuspid regurgitation. Moderate tricuspid regurgitation was observed in two patients of these, one patient was a small infant who had a VSD complicated by pulmonary hypertension. The other patient had a VSD with a mitral cleft, pulmonary hypertension, and Down's syndrome. The incised tricuspid valve was resuspended by solely running sutures. In conclusion, detachment of the tricuspid valve is a safe and useful method for adequate exposure of a VSD. However, this method should be avoided in patients with Down's syndrome and in small infants. Furthermore, repair of the incised valve should not be performed using only running sutures. PMID- 7488780 TI - Transmyocardial laser revascularization. AB - In transmyocardial laser revascularization a series of channels are created between the left ventricular cavity and the myocardium. The operation is usually performed in isolation via a left thoracotomy without cardiopulmonary bypass, but has been done in combination with coronary artery bypass grafting. Angina pectoris with coronary artery anatomy unsuitable for either angioplasty or bypass grafting is currently the most common indication for this procedure. PMID- 7488784 TI - Tetralogy of Fallot with an additional muscular inlet type ventricular septal defect--repair with split patch technique. AB - An 8-year-old girl with tetralogy of Fallot (TF) and double ventricular septal defects (VSDs) was reported. The additional VSD was a muscular inlet type. A split patch technique was used to cover both VSDs during the total correction. Follow-up echocardiogram demonstrated no residual shunt nor significant tricuspid valve regurgitation. The patient is doing well 2 years after operation. PMID- 7488783 TI - Aortic spoon-jaw clamp for aorto-saphenous vein anastomosis. AB - In the course of coronary bypass operations the proximal anastomosis of the saphenous vein to the aorta is commonly done either with the aorta clamped completely or by using a partially occluding clamp. While under usual circumstances, either method is feasible; under other conditions, such as in the presence of partial calcification, heavy atherosclerotic deposits, or in reoperations, it is preferable not to perform either of the above methods. The purpose of this article is to present an instrument designed to allow performance of the proximal anastomosis without application of any aortic clamping in the following manner. A relatively healthy spot on the ascending aorta is selected. Under protection of a mattress suture, a stab wound is made on the aortic wall, and the lower jaw of the instrument is inserted through it into the aortic lumen. A spot is selected for the anastomotic site, the instrument is placed to that area, and its handle is closed. This effectively isolates a 10 x 8 mm area of the aortic wall from the blood flow. An opening is made in the area protected by the clamp. With the jaws of the clamp closed the vein-to-aorta anastomosis is completed as usual. PMID- 7488782 TI - Effects of dynamic cardiomyoplasty on right ventricular function. AB - This study was planned to document the right ventricular function immediately after cardiomyoplasty. Right ventricular volumes were calculated with the use of fast response thermistor. Right ventricular end-diastolic volume was reduced from 91 +/- 8 mL/M2 to 75 +/- 7 mL/M2 (p < 0.05) and right ventricular end-systolic volumes increased from 51 +/- 5 mL/M2 to 59 +/- 4 mL/M2 24 hours after the operation. Central venous pressure was raised from 2 +/- 1 to 9 +/- 3 mm H2O. These changes were interpreted as impairment of the right ventricular diastolic function. Cardiac index raised from 1.8 +/- 0.5 L/M2 per minute to 2.7 +/- 0.3 L/m2 per minute within a period of 7 days (p < 0.05). As a result of this study, we concluded that preoperatively normal right ventricular function deteriorates after the cardiomyoplasty. In the intensive care unit volume balance must be carefully monitored to avoid creating right ventricular dysfunction. PMID- 7488781 TI - Clinical results of dynamic cardiomyoplasty. AB - Thirty-five cardiomyoplasty procedures were performed in five patients with dilated and 30 patients with ischemic cardiomyopathies. Russian-made cardiomyostimulators (CMS) were implanted in these patients. Twenty-one patients underwent a one-step procedure using a left thoracotomy for cardiomyoplasty, while in 14 patients, a two-step procedure that included a mid-line sternotomy was carried out. During the average follow-up period of 9 +/- 2.1 months, studies were carried out that showed in survivors improved clinical functional status and decreased heart failure symptoms. Evidence of revascularization of the ischemic myocardium from the latissimus dorsi muscle (LDM) wrap was obtained by angiography and radioactive scintigraphy. In five patients, the CMS had to be removed due to complications, and in all these patients, their clinical functional status deteriorated following the cessation of cardiomyostimulation. PMID- 7488785 TI - Excision of papillary fibroelastoma arising from the septal leaflet of the tricuspid valve. AB - A case of a papillary fibroelastoma (PFE) arising from the tricuspid valve was reported. It was incidentally detected by two-dimensional transthoracic echocardiography. Prior to 1977, these tumors were exclusively found at postmortem examination. This is only the fourth reported case of a tricuspid valve PFE found by echocardiography, treated by excision, and with tricuspid valvuloplasty preserving the native valve. PMID- 7488786 TI - New technique for retrograde cerebral perfusion during arch aneurysm repair. AB - Many techniques are used to reduce brain damage during surgery for dissecting aneurysms of the ascending aorta and arch. Recently, new techniques of protection were proposed, consistent with hypothermic circulatory arrest in association with retrograde cerebral perfusion via superior vena cava. We propose a simple, time saving method, which does not require any manipulation of the heart. We use a multilumen cannula for cardioplegia (D 860-DIDECO FUNDARO') with pressure transducer. This cannula is inserted in superior vena cava by means of a simple purse-string, and linked to the arterial line with a "Y" derivation, allowing retrograde perfusion of the brain and monitoring the perfusion pressure at every moment. The superior vena cava placed downstream from the cannula is closed by a small vascular clamp, to avoid blood reflux in the right atrium. This method is time- and money-saving, is readily available, and can be prepared whenever necessary, also in the middle of the surgical procedure. PMID- 7488787 TI - Aortic coarctation, aortic valvular stenosis, and coronary artery disease: combined one-stage surgical therapy operation. AB - The presentation of simultaneous severe coarctation of the descending aorta and severe aortic valve disease is uncommon. We describe the management of simultaneous association of aortic coarctation, aortic valve disease, and ischemic cardiomyopathy and describe a one-stage surgical approach for the correction of all pathologies. After performing the aortic valve replacement and myocardial revascularization, coarctation was solved by means of a prosthesis between the ascending aorta and the abdominal aorta. There were no evidence of myocardial ischemia during exercise testing and the blood pressure is normal. PMID- 7488788 TI - Variations in the papillary muscles of the normal mitral valve and their surgical relevance. AB - A mid-mitral plane passing through the middle of the aortic and mural leaflets divides the chordopapillary support of the mitral valve into anterolateral and posteromedial halves. The papillary muscles of the mitral valve were studied in 100 human autopsy hearts collected at random. The anterolateral papillary support had 1 belly in 67 hearts, 2 in 27, 3 in 4, 4 in 1, and 5 in 1 heart. Likewise, the posteromedial papillary support had 1 muscle belly in 50 hearts, 2 in 36, 3 in 11, and 4 in 3. The single papillary muscles were conical, mammillated, flat topped, grooved, stepped, wavy, arched, sloped or saucerized. When there were two bellies they presented a two tiered, interlinked, parallel, arched, V, Y, or H configuration. Three papillary muscles formed a parallel, interlinked or arched arrangement; or two bellies were interlinked or formed a two tiered arrangement with the third belly separate. When four or five bellies existed, they were parallel or interlinked. In the anterolateral and posteromedial group, the papillary muscle bellies were mostly intraluminal in 14% and 11%, mostly intraluminal with the tip anchored in 19% and 28%, equally sessile and intraluminal in 54.5% and 41.5%, mostly sessile in 12.5% and 19.5%, respectively. In the anterolateral group 19% of papillary muscle bellies arose from the upper third of the ventricle, 79.5% from middle third, and 1.5% from lower third. The corresponding figures for posteromedial group are 6%, 92.5%, and 1.5%, respectively. Four to 22 chordae originated from the anterolateral papillary group, ending in 14 to 72 chordal insertions into the corresponding half of the valve. Likewise, 2 to 18 chordae arose from the posteromedial papillary group ended in 12 to 80 leaflet insertions. The chordae in each group are best considered in toto as a fan. The configuration of the fan is unique in each heart. Imaging techniques need to be refined to outline these variations more precisely. The relevance of chordopapillary variations in rheumatic heart disease, reparative procedures, papillary muscle dysfunction, mitral valve prolapse, mitral valve replacement, and use of mitral valve homograft for mitral/tricuspid replacement is discussed. PMID- 7488789 TI - Surgical management of mobile aortic atheroma diagnosed by intraoperative transesophageal echocardiography. PMID- 7488790 TI - Osseus metaplasia with functioning marrow in a calcified aortic valve. PMID- 7488791 TI - A simple method to protect a fragile aorta. PMID- 7488792 TI - Warm blood cardioplegia. PMID- 7488793 TI - Laser hazards and safety. AB - Lasers are a key component of the vision practitioner's arsenal of care, and provide a wonderful new frontier of diagnostic and therapeutic options. In light of their expanded utilization, it is crucial that eye care practitioners have a thorough understanding of potential laser hazards and the regulations that establish guidelines for their safe application. This review describes the American National Standards Institute (ANSI) standards applicable to the safe use of lasers (Z 136.1) and the safe use of lasers in health care facilities (Z 136.3). Laser hazard classification, the organization of a laser safety program, and administrative and procedural controls are included. PMID- 7488796 TI - Laser-tissue interactions. AB - Current ophthalmic laser treatment can be described on the basis of its effect on tissue. Laser variables such as wavelength, spot size, power, and mode of delivery are all controllable, but are dependent on the transparency, pigmentation, and water content of the tissue involved. This discussion introduces the clinician to these variables, while describing specific laser tissue interactions. PMID- 7488795 TI - Integrating lasers into clinical practice. PMID- 7488797 TI - Complications of anterior segment laser procedures. AB - The majority of complications associated with laser procedures can be minimized or avoided through proper technique. The astute clinician is aware of all potential complications and is prepared to manage any undesired effects promptly and correctly. This review discusses how to minimize the risk of significant complications and describes the appropriate treatment protocols for complications associated with specific anterior segment laser procedures. PMID- 7488798 TI - Photorefractive and phototherapeutic procedures. AB - The excimer laser will have a tremendous impact on the future of eye care. This article covers the basic concepts of photorefractive and phototherapeutic keratectomy. Preoperative, operative, and postoperative techniques are reviewed. Common postoperative complications are discussed. This article also reviews clinical data and future trends in excimer technology. A comparison of photorefractive keratectomy with radial keratotomy is also provided. PMID- 7488794 TI - Lasers and liability. AB - Ophthalmic laser procedures are not a significant cause of liability claims. When claims are brought, they usually allege negligence, breach of the doctrine of informed consent, or both. The most likely cause of a liability claim is laser photocoagulation for diabetic retinopathy. Among anterior segment procedures, treatment for glaucoma is most likely to produce a claim. PMID- 7488799 TI - Capsulotomy. AB - The development of posterior capsule opacification secondary to extracapsular cataract extraction is a common occurrence. The patient complains of decreased visual acuity, glare, and sensitivity to light. The YAG laser posterior capsulotomy has become standard treatment for this anomaly: an opening is created in the opacified posterior capsule, which immediately improves visual acuity. This review describes the indications, preparation, procedure, follow-up, and complications associated with using a yttrium-aluminum-garnet (YAG) laser to perform capsulotomy. PMID- 7488800 TI - Laser therapy for open-angle glaucoma. AB - Management of open-angle glaucoma is complex and consists of medications, laser therapy, and conventional surgery. Laser therapy is a vital component of management, with the most common procedure being a trabeculoplasty, which reduces intraocular pressure by increasing aqueous outflow. This review discusses the indications, contraindications, and specific techniques for laser trabeculoplasty. The use of holmium sclerostomy is also discussed. With recent advances in laser and fiberoptic technology, sclerostomy is proving to be a simpler procedure than conventional filtering surgery, while providing comparable effectiveness. PMID- 7488801 TI - Laser therapy and angle-closure glaucoma. AB - All forms of primary and secondary pupillary block angle-closure glaucoma result in an increased resistance to aqueous flow at the pupil margin. As the pressure in the posterior chamber increases, the iris bows forward until the trabecular meshwork is occluded and aqueous outflow is interrupted. Most pupillary block glaucomas are ocular urgencies and require immediate medical therapy to lower intraocular pressure. The definitive treatment for pupillary block is laser therapy. By creating a hole in the iris, the pressure in the anterior and posterior chambers is equalized, allowing the angle to remain open. This discussion reviews the indications, contraindications, and techniques involved in the laser management of angle-closure glaucomas. PMID- 7488802 TI - Commentary on neonatal ECMO: a North American and Scandinavian perspective. AB - Randomized clinical trials have become the cornerstone of clinical decision making in neonatal medicine. The need to establish the safety and efficacy of different treatment regimens by means of an unbiased approach has led to the formation of state-agency supported and voluntary national and international networks. These neonatal networks have greatly facilitated the organization of randomized multicenter trials. Most studies aimed to evaluate specific treatment alternatives for common disorders (e.g. surfactant therapy for RDS and iv gammaglobulins for prevention of nosocomial infections are particularly well suited for a randomized clinical trial). However, appraisal of alternative therapies for rare conditions with excessive mortality and high risk for later sequelae is limited by a number of practical and ethical considerations. In view of the complexity of the problems involved it is hardly surprising that only one recent head-to-head trial of conventional therapy versus ECMO has to our knowledge been published (39). In this investigation 28 full-term infants fulfilling standard ECMO criteria were randomly assigned to be transported for ECMO or to receive conventional treatment available on-site. Fourteen of the 15 infants (93%) referred for ECMO survived compared with 7 of 13 (53%) treated conventionally. The long-term outcome was comparable in both groups. These data support previous reports of significantly better survival following ECMO treatment. However, the notion of > 80% mortality on continuation of conventional therapy in this critically ill population may need to be reassessed. HFO is today part of "conventional" therapy in many centers, surfactant is used routinely and a combination of NO and HFO is being evaluated in several clinical trials (57).(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7488803 TI - Diagnosis and presentation of rickets. PMID- 7488804 TI - Effect of the method of breast feeding on breast engorgement, mastitis and infantile colic. AB - This exploratory study compared the effect of two methods of breast feeding on breast engorgement, mastitis, infantile colic and duration of breast feeding. An opportunity sample of subjects was assigned either to the experimental group (prolonged emptying of one breast at each feed) (n = 150) or to the control group (both breasts equally drained at each feed) (n = 152) and both groups were followed prospectively to 6 months after delivery. The experimental group had a lower incidence of breast engorgement in the first week (61.4% versus 74.3%; p < 0.02) and colic over the first 6 months (12% versus 23.4%; p < 0.02). There was no significant difference between the two groups in the incidence of mastitis over 6 months and the length of breast feeding (16.5 +/- 10.3 weeks versus 17.5 +/- 10 weeks experimental versus control group). The majority of mothers in the experimental group (63%) felt it necessary to offer the second breast at the end of a feed to satisfy their infant's hunger. The "perceived insufficient milk supply syndrome" was the main reason given for cessation of breast feeding in both groups. This study provides data to advise nursing mothers about these two methods of breast feeding. PMID- 7488805 TI - Weaning practices in children up to 19 months of age in Madrid. AB - The weaning practices of 344 children in Madrid, aged 3-19 months, were investigated by controlled interview of their mothers or caretakers. The results were compared with national and international guidelines for the introduction of complementary foods; there are no Spanish guidelines. We found only minor differences between the practices in three different socioeconomic groups. Generally, mothers in Madrid followed national and international guidelines well. However, more attention should be paid to aspects such as the timing of the introduction of the first complementary food, the nature of gluten, the addition of salt and sugar to baby meals and the introduction of potentially allergenic foods. Also, manufacturers of baby foods that are sold in Spain could play a useful role by reducing the number of ingredients used in products advised for the initial phase of weaning and by avoiding the use of salt and sugar in their products. PMID- 7488806 TI - Selenium in German infants fed breast milk or different formulas. AB - At birth and at 4 months of age, selenium (Se) values of 129 term infants on three different diets were determined: 50 infants were breast fed (HM), 44 received formula based on cow's milk (F) and 35 were fed "hypoallergenic formula" (PHF) (partially hydrolysed whey protein). The Se status of a group of twins (n = 12) fed "hypoallergenic formula" was compared with the respective group of singletons. All infants had low plasma Se values during early infancy. The plasma Se of breast-fed infants remained stable (plasma Se 43 +/- 8 ng/ml at birth and at 4 months), whereas plasma glutathione peroxidase (GSH-Px) decreased (birth: 107 +/- 29 U/l; 4 months: 62 +/- 11 U/l). The formula-fed infants showed a reduction in plasma Se levels from birth to 4 months (38 +/- 10 ng/ml and 29 +/- 9 ng/ml, respectively). The decrease was even more pronounced in infants fed the "hypoallergenic formula". This group presented the lowest Se values (plasma Se 39 +/- 9 ng/ml at birth; 20 +/- 6 ng/ml at 4 months). Renal excretion of Se was found to be lower in the formula-fed infants (F and PHF) compared with the HM group. There was a significant correlation between plasma and urinary Se (r = 0.62, p = 0.0001). Urinary Se (microgram Se/g creatinine) appeared to be a good indicator of Se intake. Measurements of urine Se might be used as a screening method for the estimation of the Se supply. Weight and length increases in all infants were within the normal range. There were no differences between the different feeding groups. PMID- 7488807 TI - Bulging fontanelle after supplementation with 25,000 IU of vitamin A in infancy using immunization contacts. AB - To evaluate the safety of vitamin A supplementation in early infancy using DPT/OPV immunization contracts, a double-blind, randomized, placebo-controlled trial was conducted in Bangladesh. One hundred and sixty-seven infants received three doses of either 25,000 IU of vitamin A or a placebo at about 6.5, 11.8 and 17.0 weeks of age. Trained physicians examined each of the infants on days 1, 2, 3 and 8 after supplementation. Nine infants (10.5%) supplemented with vitamin A had episodes of bulging of the fontanelle compared with two infants (2.5%) in the placebo group (p < 0.05). Twelve of the 14 episodes occurred in infants supplemented with vitamin A. Of these 12 episodes, none occurred with the first dose, 3 occurred with the second and 9 with the third dose. The higher incidence of bulging of the fontanelle in the vitamin A group relative to the placebo group and its temporal association with the vitamin A doses are suggestive of a causal association. The finding that increased numbers of vitamin A doses were associated with a higher probability of bulging of the fontanelle suggests a cumulative effect. PMID- 7488808 TI - Role of amylase-treated, energy-dense liquid diet in the nutritional management of acute shigellosis in children: a controlled clinical trial. AB - To evaluate if an energy-dense porridge liquefied by amylase-rich flour (ARF) from germinated wheat increased the calorie intake in children with acute shigellosis, we studied 66 children, aged 6-35 months, in a randomized, controlled clinical trial. Children were randomized to receive either an energy dense porridge liquefied with ARF (group 1), a thick unaltered porridge (group 2) or a porridge diluted with water (group 3) to a similar viscosity as that in group 1. Mean +/- SD calorie intakes (kJ/kg/day) from the porridges were 280 +/- 113, 167 +/- 100 and 151 +/- 80 in groups 1, 2 and 3, respectively (p = 0.006, ANOVA). Total energy intakes (mean +/- SD) from the study diet and other food sources were 469 +/- 151, 377 +/- 121 and 351 +/- 100 kJ/kg/day, respectively (p = 0.006, ANOVA). Intake of breast milk was similar in all groups. Using multiple regression analysis the effect of ARF-treated energy-dense porridge in increasing the calorie intake persisted after adjusting for a number of confounders, such as age of the child, isolation of Shigella dysenteriae type 1 and fever. The results of this study suggest that ARF-treated porridge increases energy intake in infants and young children during acute shigellosis. This feeding approach may be useful in preventing malnutrition following dysentery due to shigellosis. PMID- 7488809 TI - Prognostic factors in childhood bacterial meningitis. AB - The purpose of this study was to identify possible risk factors associated with a poor prognosis in childhood bacterial meningitis. We also analysed the influence of duration of symptoms and prehospital antibiotic therapy on outcome. Ninety-two children aged 1 month to 13.8 years were included, of whom 4 died (4.3%) and 14 (15.2%) experienced permanent neurological sequelae. Hearing impairment was the most frequent sequela and was strongly associated with the length of history. Multiple logistic regression revealed duration of symptoms > 48 h, pre-hospital seizures, peripheral vasoconstriction, < 1000 x 10(6)/l leucocytes in cerebrospinal fluid and temperature < or = 38.0 degrees C on admission as risk factors independently associated with later death or sequelae. There was no association between pre-hospital oral or parenteral antibiotic therapy and outcome. These risk factors may be of value in selecting patients for more intensive therapy and in identifying possible candidates for new treatment strategies. PMID- 7488810 TI - Interleukin-6 in cerebrospinal fluid of patients with central nervous system infections. AB - Interleukin(IL)-6 levels were measured in cerebrospinal fluid (CSF) and serum samples from pediatric patients with central nervous system (CNS) infections by means of an enzyme-linked immunosorbent assay. Mean IL-6 concentrations in CSF samples from patients with bacterial meningitis (49,017 +/- 44,730 pg/ml) were significantly higher than those in patients with aseptic meningitis (1076 +/- 1572 pg/ml) or encephalitis (409 +/- 835 pg/ml). In aseptic meningitis and encephalitis, IL-6 levels in serum were within the lower ranges (< 100 pg/ml), in contrast with the highly elevated levels found in bacterial meningitis (14,332 +/ 18,385 pg/ml). In 5 of the 15 patients with encephalitis, elevated levels of IL 6 were observed in the initial CSF samples despite normal findings of routine CSF examinations. Also, sequential CSF samples revealed that there was an increase in the CSF cell count in two of the five patients. These results validated the potential of measuring IL-6 in CSF samples for the purpose of providing additional information on routine laboratory test results. PMID- 7488811 TI - Intermittent treatment with inhaled steroids for deterioration of asthma due to upper respiratory tract infections. AB - Upper respiratory tract infection (URTI) is a common cause of deterioration of asthma in children. We investigated if inhaled steroids (budesonide), started early after URTI, could reduce asthma. Thirty-one children, 3-10 years of age, with deterioration during URTI participated. The study design was double-blind, crossover and placebo-controlled. Peak-expiratory flow (PEF) and symptom scores were recorded. Four treatment periods of 9 days, two with budesonide and two with placebo, were planned. Treatment was started at the first sign of URTI. Budesonide/placebo was given by Turbuhaler at 0.2 mg qid for 3 days, tid for 3 and bid for the last 3 days. Twenty-two children completed 67 periods. Eleven visited the emergency room, only three during budesonide therapy. Five received oral steroids and two where admitted to hospital, all receiving placebo. Symptom scores were not significantly lower during budesonide treatment. PEF, both morning and evening, was significantly higher during budesonide than placebo (p = 0.015 and p = 0.022). Inhaled budesonide can attenuate exacerbation of URTI induced asthma. PMID- 7488812 TI - Short versus long initial prednisone treatment in steroid-sensitive nephrotic syndrome in children. AB - A total of 184 children aged, 13 months to 11 years, suffering from their first attack of steroid-responsive nephrotic syndrome were included in a randomized study. They were treated according to three treatment protocols. All children received 1-2 mg of prednisone/kg body weight/day (up to 80 mg daily) for 4 weeks, and thereafter 1 mg/kg body weight/48 h for the next 4 weeks. Treatment was discontinued at this point in 44 children (protocol A); in 68 (protocol B) the dose was reduced by 25% each week, tapering off to 0 at the end of the third month, while in 72 children (protocol C), after the first 2 months of initial treatment the dose was reduced by 25% each month and tapered off to 0 by the end of the sixth month. All patients completed a 2-year follow-up period after withdrawal of prednisone. Treatment results were expressed as: percentage of children relapse-free within the first 6 months and 2 years after withdrawal of treatment, and average number of relapses per patient per year. The best results were obtained in children who had been treated for 6 months; 65.3% of them remained relapse-free within the first 6 months and 50% over the entire 2-year follow-up period; the number of relapses per patient per year in this group was 0.49. The respective values for children treated 2 and 3 months were: 36.4% and 32.4% for the 6-month period; 27.3% and 20.6% for the 2-year period; the numbers of relapses per patient per year were 0.79 and 0.77, respectively.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7488813 TI - Early growth is not increased in untreated moderately severe 21-hydroxylase deficiency. AB - The pretreatment growth of 1 British and 14 Swedish children with late (2-7 years) diagnosis of 21-hydroxylase deficiency (21OHD) was studied. The latter group included all patients diagnosed in Sweden after 1986. Twelve had mutations of the 21-hydroxylase gene that are generally associated with moderately severe ("simple virilizing") forms of 21OHD, one had a severe ("salt-losing") and one a mild ("non-classical") form. The British girl was followed from 4 months of age. She had grossly elevated levels of 17 alpha-hydroxyprogesterone, androstenedione and testosterone in blood, but her parents refused treatment until she was 4 years of age. None of the 15 children showed any significant increase in growth or progress of virilization until after 18 months of age. These observations indicate that growth during the first 1.5 years is not very sensitive to androgens. Thus glucocorticoid replacement during the first year of life should be kept to a minimum to avoid over-treatment. PMID- 7488814 TI - Adult height in women with early-treated congenital adrenal hyperplasia (21 hydroxylase type): relation to body mass index in earlier childhood. AB - To investigate if obesity, as judged by increased body mass index (BMI), during childhood is associated with impaired adult stature in patients with 21 hydroxylase-type congenital adrenal hyperplasia (CAH), a retrospective analysis was carried out on outpatient records of 30 girls with CAH who had reached adult height. Height SD score for age (HtSDS(ca)), HtSDS for bone age (HtSDS(ba)), BMI and steroid dosage in early childhood (3.2-4.6 years) and later childhood (7.2 9.1 years), were compared with adult HtSDS (HtSDS(adlt)), adult HtSDS less mid parental HtSDS (HtSDS(adlt)-HtSDS(mp)), predicted adult height SDS (HtSDS(pdct)), adult height SDS less predicted adult HtSDS (HtSDS(adlt)-HtSDS(pdct)) and age at menarche. Mean (SD) for HtSDS(adlt) was -1.13 (1.05), mean HtSDS(pdct) -0.12 (0.9) and mean age at menarche 13.5 (1.9) years. BMI in childhood was not correlated with HtSDS(adlt) but showed negative correlations with HtSDS(adlt) HtSDS(mp) (r = -0.43; p < 0.02) and HtSDS(adlt)-HtSDS(pdct) (r-0.45; p < 0.02). BMI in later childhood showed negative correlations with HtSDS(adlt)-HtSDS(pdct) (r = -0.61; p < 0.001) and age at menarche (r = -0.63; p < 0.001). We conclude that in girls with early-treated CAH, a high BMI during early childhood is associated with loss of genetic height potential, and in later childhood with over-prediction of adult height and early menarche. PMID- 7488815 TI - Plasma growth hormone response to growth hormone-releasing hexapeptide (GH-RP-6) in children with short stature. AB - Eighteen children with short stature were evaluated for growth hormone (GH) reserve after pharmacological tests and a single iv injection of GH-RP-6. These children were divided into two groups: 10 were diagnosed as having idiopathic GH deficiency by classical stimulation tests (group A) and the remaining 8 (group B) were considered growth-retarded children with normal GH secretion, following conventional stimulation, but reduced endogenous GH secretion. The results were compared with a group of 12 normal children. As a group, patients in group A showed a lower GH response to GH-RP-6, while patients in group B had a similar response as normal controls. However, on an individual basis, a considerable degree of overlapping in responses among the three groups was evident. These data indicate that, on an individual basis, GH-RP-6 testing is not of diagnostic value in children suspected of having idiopathic GH deficiency. PMID- 7488816 TI - Frequency, clinical and laboratory features of thyroiditis in girls with Turner's syndrome. The Italian Study Group for Turner's Syndrome. AB - A total of 478 patients, mean age 15.5 (3.6-25.3) years, suffering from Turner's syndrome, were studied in order to determine the frequency of autoimmune thyroiditis, which is defined as the presence of antithyroid antibodies (AT-Ab) and typical ultrasound findings. We found 106 (22.2%) patients positive for AT-Ab and of those 49 (10%) also had positive ultrasound findings, and were therefore considered to be affected by thyroiditis. This frequency is significantly higher (p < 0.001) than that seen in the normal population. Goitre was detected on clinical examination in only 16 (33%) and by ultrasound in 19 (39%) patients. Hormonal evaluation showed that 17 patients were euthyroid, 27 had compensated hypothyroidism, 2 were hypothyroid and 3 were in a hyperthyroid phase. Clinical signs or symptoms of hypothyroidism were absent in all hypothyroid patients. In patients with thyroiditis, neither a higher frequency of malformations and autoimmune diseases nor a correlation with karyotype, oestrogens or growth hormone therapy was found. PMID- 7488817 TI - Porcine surfactant replacement therapy in newborns of 25-31 weeks' gestation: a randomized, multicentre trial of prophylaxis versus rescue with multiple low doses. The French Collaborative Multicentre Study Group. AB - The aim of the study was to determine if prophylaxis with multiple low doses of porcine surfactant would increase survival, without bronchopulmonary dysplasia, compared with rescue therapy, for respiratory distress syndrome in newborns of 25 31 weeks' gestation. Compared with rescue therapy (n = 122), prophylaxis (n = 134) decreased the need for oxygenation and ventilatory support within 3-72 h. It did not, however, increase survival without bronchopulmonary dysplasia (60% versus 46%) (odds ratio (OR) = 1.53, 95% confidence interval (CI) = 0.90-2.61). Furthermore, prophylaxis decreased the incidence of severe peri-intraventricular haemorrhage (3% versus 16%) (OR = 0.28, 95% CI = 0.09-0.84) and retinopathy of prematurity (2% versus 11%) (OR = 0.18, CI = 0.04-0.78). We conclude that prophylaxis did not increase survival without bronchopulmonary dysplasia. The decreased incidence of severe peri-intraventricular haemorrhage and retinopathy of prematurity after prophylaxis requires further study. PMID- 7488818 TI - Effect of surfactant on nitroblue tetrazolium reduction of polymorphonuclear leucocytes stimulated with type Ia group B streptococci. AB - Activation of polymorphonuclear leucocytes (PMN) was investigated after incubation of adult human PMN and group B streptococci (GBS) type Ia with a type specific polyclonal antiserum and a modified porcine surfactant (Curosurf). The level of oxidative metabolism of PMN was studied using a micromethod modification of the nitroblue tetrazolium (NBT) reduction test. GBS alone did not stimulate significant oxygen metabolite release from PMN, and incubation of PMN with surfactant alone resulted in decreased NBT reduction. After opsonization of GBS with a specific antibody, PMN were activated and the increased oxygen metabolite release was not suppressed when surfactant was added to the system. We conclude that the encapsulated GBS strain investigated needs opsonization with specific antibody to increase oxidative metabolism of PMN, and that incubation of PMN and opsonized GBS with surfactant does not interfere with NBT reduction. PMID- 7488819 TI - Birth asphyxia: incidence, clinical course and outcome in a Swedish population. AB - A total of 42,203 live infants were born in Goteborg in 1985-1991, and 292 term infants had Apgar scores < 7 at 5 min. Infants with congenital malformations, infections and opioid-induced respiratory depression were excluded and thus 227 infants were included in the birth asphyxia group, which formed the basis of this retrospective study. Clinical signs of mild, moderate or severe hypoxic-ischemic encephalopathy (HIE) were present in 65 infants, and in another 10 infants, sedated and on controlled ventilation, HIE was assumed but grading was not possible. The incidences of Apgar scores < 7 at 5 min, birth asphyxia and birth asphyxia with HIE were 6.9, 5.4 and 1.8 per 1,000 live born infants: 95% of infants resuscitated with bag and mask ventilation only, did well, compared with 1 of 11 in whom resuscitation included adrenaline. Seizures occurred in 27 of 227 infants, beginning in 18 infants within 12 h of birth. Small-for-gestational-age (SGA) infants were overrepresented in the birth asphyxia group but not in the birth asphyxia-HIE group. All infants with severe HIE died or developed neurological damage. Half of the infants with moderate, and all of the infants with mild, HIE were reported to be normal at 18 months of age. A total of 0.3 per 1,000 live born infants died and 0.2 per 1,000 developed a neurological disability related to birth asphyxia. The disabilities were dyskinetic (4), tetraplegic (2), spastic diplegic (2), cerebral palsy and mild neuromotor dysfunction (1). The relatively low incidences of birth asphyxia and HIE were probably due to effective antenatal care. PMID- 7488820 TI - Low levels of plasminogen in cerebrospinal fluid after intraventricular haemorrhage: a limiting factor for clot lysis? AB - The aim of this study was to measure plasminogen in the cerebrospinal fluid (CSF) of control neonates with no infection or haemorrhage and in infants who had suffered intraventricular haemorrhage (IVH). A chromogenic substrate method was used. The 16 reference infants had a median CSF plasminogen level of 0.74% of that of normal adult plasma (range 0.17-1.1%). The 11 infants with IVH had a median CSF plasminogen level of 0.55% of normal adult plasma (range 0-4.4%). Six of the IVH infants went on to develop permanent hydrocephalus despite the use of intraventricular plasminogen activators. Endogenous fibrinolysis and the potential for fibrinolytic treatment in the CSF may be limited by low concentrations of plasminogen, and administration of recombinant plasminogen may assist attempts to clear intraventricular blood clots. PMID- 7488821 TI - Magnetic resonance imaging in children with kernicterus. AB - The magnetic resonance (MR) findings of three children with severe neonatal jaundice who were suspected to be suffering from kernicterus were examined. In all children high intensity areas in the globus pallidus were demonstrated bilaterally on T2-weighted imaging. The posteromedial border of the globus pallidus was shown to be the most sensitive area to kernicterus by MR findings. PMID- 7488823 TI - Vitamin D deficiency rickets in breast-fed infants presenting with hypocalcaemic seizures. AB - At the Aga Khan Hospital (AKUH), 65 infants presented with hypocalcaemic seizures, subsequently found to have rickets. Forty-six infants less than 6 months were totally or predominantly breast fed. In a subgroup of 15 mothers and their infants, we found very low plasma levels of 25(OH) vitamin D of < 5 micrograms/l and 7.5 +/- 3.3 micrograms/l, respectively. Neither mothers nor infants received vitamin D supplementation. Maternal vitamin D deficiency and non supplementation in the infants were the likely causes of rickets in our patients. Prophylactic vitamin D 400 i.u. administered to infants up to 2 years and 800 i.u. to women in pregnancy and during lactation is recommended to prevent vitamin D deficiency. PMID- 7488824 TI - Recombinant granulocyte-macrophage colony-stimulating factor in the treatment of HIV-related leucopenia. PMID- 7488825 TI - Gastric aspirate volume at birth as an indicator of congenital intestinal obstruction. PMID- 7488822 TI - Prevention of rickets and vitamin D intoxication in China. PMID- 7488826 TI - Aldolase B A149P mutation and hereditary fructose intolerance are not associated with sudden infant death syndrome. PMID- 7488827 TI - Postoperative chylous ascites. AB - The case of an infant, aged 2 months and 23 days, with chylous ascites after nephrectomy is reported. Nephrectomy was performed because of a large hydronephrosis. Chylous ascites was treated with medium-chain triglycerides, diet and cholestyramine. After 3 days the volume of the abdomen was reduced and on day 8 chylous ascites was not visible clinically or ultrasonographically. PMID- 7488829 TI - Isolated congenital tuberculosis otitis in a preterm infant. AB - We report an unusual case of localized congenital tuberculosis otitis in a preterm infant. Unlike disseminated congenital cases, the manifestations of localized otitis are associated with a triad of signs: (i) regional lymphadenopathy in the absence of typical systemic features of tuberculosis; (ii) delayed onset of presentation; and (iii) refractory otitis unresponsive to conventional antimicrobial agents. The need for greater diligence in looking for neonatal tuberculosis is emphasized, especially in an ethnic or socioeconomic environment where the disease is prevalent. PMID- 7488828 TI - Sulphasalazine desensitization in a paediatric patient with juvenile chronic arthritis. AB - Sulphasalazine is an effective drug for the treatment of rheumatoid arthritis in adults. In paediatric patients, the drug has been used to treat inflammatory bowel disease and is currently under investigation for the treatment of juvenile chronic arthritis. Although sulphasalazine has a rather low incidence of serious side effects, one of the most common is skin rash, thought to be an allergic reaction. In adults, sulphasalazine desensitization programmes have proven to be effective for the treatment of this side effect. We present the case of a 7-year old boy suffering from HLA-B 27 positive juvenile chronic arthritis. After initiation of treatment with sulphasalazine he developed an allergic skin rash, but tolerated the drug well after completion of a desensitization programme. To our knowledge, this is the first report of a paediatric patient with juvenile chronic arthritis successfully desensitized with sulphasalazine. PMID- 7488831 TI - Defining childhood obesity: the relative body mass index (BMI). European Childhood Obesity group. PMID- 7488830 TI - A special case of congenital adrenal hypoplasia and acute bilateral infantile striatal necrosis. AB - Isolated mineralocorticoid deficiency is described in a 5-week-old boy. The deficiency progressed to general adrenal insufficiency during the boy's first year of life. The family history suggested X-linked inheritance. At 18 months of age the patient developed acute bilateral infantile striatal necrosis, which might suggest a possible relationship between both entities. PMID- 7488832 TI - Candida antigen and antibody assays. PMID- 7488833 TI - Histopathological features of liver disease. PMID- 7488834 TI - Hospice movement in the world and its future in Croatia. PMID- 7488835 TI - Papers published by members of the Croatian Academy of Medical Sciences, registered in international bibliographic data bases in the period 1986-1990. AB - Scientific productivity of the CAMS members was analyzed according to the number of papers published in periodicals covered by international Medline and Science Citation Index data bases from 1986 to 1990. Results showed the mean scientific productivity per CAMS member, and the relationship between scientific productivity and age, field of medicine or respective biomedical discipline. The pattern of scientific productivity was also found to change, when analyzed according to Medline or SCI data bases in separate. PMID- 7488836 TI - Differences between male and female breast cancer. I. Epidemiological features. AB - The authors' interest was focused on the direct determination of differences between the salient features of breast cancer in men and women in a large number of patients. Various epidemiological features were analyzed in a group of 386 male and 21,491 female breast cancer patients registered in the Cancer Registry of the Republic of Croatia from 1968 until 1988. This was a population-based study, and all male and female breast cancer cases recorded during the mentioned period were included in the study. During this 21-year period, the total incidence rate of breast cancer in men was about 0.83/100,000, and the mortality 0.2,/100.000. In the same interval, in women the rates doubled from 30 to 60/100,000, and 13 to 29/100,000, respectively. The highest incidence rates were recorded in the 80-84 age group in men, and 60-64 age group in women. The mean age at diagnosis was 63.4 years in men and 58.2 years in women. The distribution in terms of TNM-stages in men was: stage I-10.10%; II-40.67%; III-32.40%; IV 16.83%; in women; I-12.19%; II-49.36%; III-27-58%; IV-10.86%. The results showed that breast cancer in men had epidemiological characteristics of a sporadic disease, occurring continuously at a specific mean frequency in the general population and being little affected by environmental impacts. Conversely, in women affected by the disease there was a tendency of continuous rise in the morbidity and mortality, which might have been due to the trigger action of one or more environmental factors.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7488837 TI - Personality traits in school children with epilepsy. AB - An alternative way of looking at epilepsy has been restricted to the observation of personality characteristics and behavioral cognitive impairments, as these are the most important secondary handicaps. The term "behavioural disorders" is connected with the problem of the development of personality. For this reason, in clinical work a sensitive and standardized psychometric instruments for measuring personality constructs are required. One of the most frequent used instruments in our child neurowork is Eysenck's Personality Inventory (EPI--junior questionnaire), based on Eysenck's theory. In this study, EPI-junior questionnaire was given to a group of 60 boys and girls aged 10-14 years with various forms of epilepsy (single partial seizures--N = 28; complex partial seizures--N = 18; typical absence--N = 8; atypical absence--N = 6. All children were receiving anticonvulsant drugs in doses within or below therapeutic limits. The possible influence of drug administration on personality characteristics of these children was not specifically analyzed for insufficient data in their medical histories. Results of personality characteristics obtained on the EPI junior test of the children with epilepsy were compared to the results of "normal" school children matched by age, sex and social conditions. Its was found (on the "extroversion-introversion" scale) that the children with epilepsy were more introverted than the control group children, which is contrary to the common clinical experience. On the other hand, there were no statistical differences between these two groups in the category of "neuroticism". Finally, the children with epilepsy had significantly higher results on the "lie scale", which indicated greater unreliability of their results obtained on EPI-junior "lie" scale as compared to the control group.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7488838 TI - Classic migraine and visual field defects. AB - A series of 116 patients with classic migraine were studied to record visual field defects. Thirty-four (29.3%) subjects had defects of the field of vision, 11 of them bilaterally, which made a total of 45 (19.4%) eyes. Downward isopter depression from above and contraction of the field of vision were the most common damages. A homonymous hemianopic defect was found in seven patients. The area of depressed vision helps to some extent in anatomical localization of the damage. Its etiology has not been fully clarified. The authors recommend more extensive clinical examinations of the visual field, i.e. follow-up of each patient with classic migraine by periodical testing of the field of vision. PMID- 7488840 TI - Prolegomena for an anti-dictatorship campaign. AB - This work represents a derivative of a greater study in which adequate values should be given to the psychological component of dictatorship and tyranny within the framework of certain social conditions, economic possibilities of societies, and political events that begin and end all political differences and chaos. Society, economy, and politics are simultaneously the feeding places and hunting ground where ambitious individuals and suggestible masses meet. This "fortunate meeting" leads to dictatorship, which wouldn't be possible without these two psychological components. In our life we have become aware of the irresistible fascination between the masses and a future dictator who uses the masses for the encouragement of his own defenses embodied in narcissism, hatred, and personal will. However, we shouldn't forget that the dictator's characteristics and aspirations represent mostly the fragments of a frustrated childhood and extreme personal outrage and weakness. The author is not on safe ground while tracing the most convincing historical background to confirm his thesis about the appearance of the dictator and dictatorship, however neither can he find a firm stand for its prevention on these phenomena. On the contrary, these shoudn't be any delusions concerning our times and Middle-European conditions, because the appearance of a dictatorship is possible always and everywhere--just the colors are changed. Therefore, it is quite necessary to lead a permanent antidictatorship campaign. Why? Because dictatorship is always possible although it can appear in different forms and can be based on shrewd illusions. Group interests become the most obvious source of a dictatorial invasion. Today, nationalism, fed by the retardate religious remnants from the past, is on the offensive.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7488841 TI - A life-threatening complication of extreme hyperkalemia in a patient on maintenance hemodialysis. AB - A 68-year-old female on two-year chronic hemodialysis for chronic renal failure due to chronic pyelonephritis, was admitted to hospital for weakness, dulled sensorium and dizziness. On examination the patient was in a state of circulatory collapse, the electrocardiogram showed an accelerated idioventricular rhythm and laboratory analysis revealed extreme hyperkalemia (K+ 10.1 mmol/l). There were no common causes of shock, such as hypovolemia, sepsis, heart failure and presence of vasodilator drugs. The patient was treated with calcium gluconate, sodium bicarbonate and sodium chloride (to oppose the effects of hyperkalemia on the cell membrane to minimize cardiac and neuromuscular toxicity), insulin and dextrose (to increase the transport of K+ from the extracellular to the intracellular compartment), and hemodialysis (to remove K+ from the body). At the end of the hemodialysis session, the patient was in a clinically good condition, blood pressure was 160/90 mm Hg and the serum K+ concentration was normal. The case appeared to suggest that extreme hyperkalemia may have direct effects on vascular resistance, causing hypotension and shock. PMID- 7488839 TI - Hyperbaric oxygen for the treatment of nonarteritic anterior ischemic optic neuropathy. AB - This clinical study tackled the problem of hyperbaric oxygen in patients with nonarteritic anterior ischemic optic neuropathy. Twenty-one patients were included in the study. The patients were divided into two groups: experimental and control. Experimental group consisted of 10 patients, who had been previously treated with corticosteroids, without significant success. In four patients with a clinical picture of PNO atrophy, medical steroid treatment failed to any significant improvement in visual acuity. In 6 patients, however, this treatment resulted in a significant improvement of both visual acuity and field of vision. These patients showed further improvement on control testing performed six month later. All patients from the control group were treated with corticosteroids per os and by local application, but without any significant improvement as revealed by visual acuity and visual field testing. Authors' results pointed the possibility of conservative treatment of patients with anterior ischemic optic neuropathy, especially those in whom steroid therapy had proved inefficient. PMID- 7488842 TI - Analysis of renal allograft frozen-sections. AB - Frozen-section analysis of renal allograft biopsy specimens is performed to permit modification of therapy during acute events in transplanted patients. Over a two-year period, out of 110 renal allograft biopsies 66 were analyzed on a frozen-section. The authors compared diagnoses obtained on a fresh-material frozen-section with those following profound specimen analysis. The accurate diagnosis was achieved in 61 (92.4%) cases, acute rejection being predominant (53 cases). Missdiagnoses referred to oxalosis (1 case), acute rejection (2 cases), acute pyelonephritis (1 case) and cortical infarction (1 case). The authors conclude that a frozen-section analysis is a useful tool in cases requiring fast decision. PMID- 7488844 TI - Genetic approaches to mechanosensory transduction. AB - Genetic approaches in several organisms provide the means of solving a previously intractable problem: characterizing the molecular foundations of the mechanical senses. In nematode mechanosensory cells, members of a novel class of epithelial ion channel subunits have been implicated as components of a mechanically gated channel. In insect mechanosensory bristles, mutations specifically defective in mechanoreceptor potentials have been identified. And in bacteria, a stretch activated channel has been molecularly characterized for the first time. Although mechanosensitivity can be a property of an isolated channel, sensory transduction in eukaryotic mechanosensory cells probably requires the interaction of several membrane and cytoskeletal components. PMID- 7488843 TI - Family of cyclic nucleotide gated ion channels. AB - A variety of different cyclic nucleotide gated ion channels have recently been identified using molecular cloning and electrophysiological techniques. Current research is focussed on the specific molecular determinants that endow these channels with their distinctive character of gating, selectivity and modulation. In some cases, it has been possible to identify the specific physiological roles of different cyclic nucleotide gated channels. Their interactions with Ca2+ and calmodulin are particularly important, and determine the specific functions these channels subserve in distinct cells. PMID- 7488845 TI - Molecular machinery of auditory and vestibular transduction. AB - The hunt for molecules that conduct mechanoelectrical transduction in hair cells has recently intensified. A hair cell's transduction apparatus adapts to sustained stimuli, and myosin I beta and myosin VIIA have been advanced as candidates for the motor that mediates this process. The identity of the transduction channel remains unknown, although a viable suggestion proposes that it belongs to the amiloride-sensitive Na+ channel family. PMID- 7488847 TI - Dual second-messenger pathways in olfactory transduction. AB - Increasing evidence indicates that inositol phosphate as well as cyclic nucleotide signalling pathways mediate olfactory transduction. Both pathways can target multiple ion channel effectors, suggesting that olfactory receptor cells serve as more than simple selectivity filters and that they possibly represent the first stage of olfactory integration. PMID- 7488848 TI - Relation of chemical structure to specificity of response in olfactory glomeruli. AB - Recent work on the mammalian olfactory system shows that sensory neurons expressing the same type of odor receptor converge their axons onto one or a few glomeruli. This nearly one-to-one correspondence between an odor receptor type and an olfactory glomerulus gives rise to the tuning specificity of bulbar mitral and tufted cells, such that individual cells are only activated by a range of odor molecules having a similar chemical structure. These findings now make it possible to analyze the molecular mechanisms involved in the functional differentiation of sensory neurons, as well as those involved in their specific connections with bulbar neurons. PMID- 7488849 TI - Pattern recognition in insects. AB - After 80 years of research, the field of insect pattern recognition is about to move from the behavioral to the neuronal level. Recent experiments on bees, ants and flies indicate that pattern recognition must be seen as the recruitment of behavioral operations that help the nervous system to solve a task using a small number of potentially simple processing steps. These may now be identified physiologically. PMID- 7488846 TI - The genetics of olfaction. AB - Our understanding of olfaction has progressed rapidly in recent years as a result of the molecular genetic approaches being used to study this sensory system in a variety of model organisms. Considerable success has been achieved in identifying proteins of the mammalian signaling system that are analogous to those present in other sensory systems. More recently, genetic selection of mutations that cause defects in olfactory function in Drosophila melanogaster and Caenorhabditis elegans has led to the identification of additional proteins that play a role in the detection of odorants. The application of genetic, electrophysiological, and molecular analyses to olfactory function in mammals is also shedding light on the mechanisms that account for sensitivity and specificity in this system. PMID- 7488850 TI - Olfactory learning. AB - Unravelling the mechanisms of learning and memory can, and should, be tackled at many levels. Discovery of the huge family of odourant receptor genes provided olfaction with 'molecular' respectability similar to that afforded to the visual system. Consequently, molecular studies have dominated the olfactory literature this past year, even to the point of providing a molecular basis of olfactory perception. Needless to say, the molecular approach favours a 'hard-wired' system; however, other results suggest that flexibility in the olfactory system provides for certain adaptations that are crucial to the biological needs of mammals. PMID- 7488851 TI - Order and disorder in auditory cortical maps. AB - Recent physiological experiments suggest that several basic receptive field properties of neurons show non-uniform spatial distributions in the primary auditory cortex of cats and primates. The spatial distribution patterns of some of these receptive field parameters are suggestive of a parallel coding scheme for processing sound information onto several superimposed cortical 'maps'. The representations of these parameters in the auditory cortex are compatible with general features of self-organizing mapping algorithms, as the spatial representations exhibit global parameter gradients with overlaid functional patchiness. Recent studies have also revealed that within a 'representational' map, the degree of local coherence varies over a wide range and that the map can contain a substantial degree of disorder in its parametric representation of sounds. Although the causes and consequences of this representational disorder are not known, it may reflect yet unresolved organizational principles in the auditory cortex. PMID- 7488852 TI - Parallel versus serial processing: new vistas on the distributed organization of the visual system. AB - Recent functional studies question the validity of the hierarchical model of organization for processing visual information in cortical areas. The results of these studies suggest that beyond the primary visual cortex (V1), information is not serially processed through successive cortical areas, but that it is simultaneously processed in several areas. The idea that visual information is functionally segregated into different, parallel channels as it circulates through V1 and V2 towards V4 and the middle temporal visual area is also challenged by recent studies that report a smaller degree of functional specialization within the visual areas than previously thought. PMID- 7488853 TI - Temporal structure in spatially organized neuronal ensembles: a role for interneuronal networks. AB - Network oscillations are postulated to be instrumental for synchronizing the activity of anatomically distributed populations of neurons. Results from recent studies on the physiology of cortical interneurons suggest that through their interconnectivity, they can maintain large-scale oscillations at various frequencies (4-12 Hz, 40-100 Hz and 200 Hz). We suggest that networks of inhibitory interneurons within the forebrain impose co-ordinated oscillatory 'contexts' for the 'content' carried by networks of principal cells. These oscillating inhibitory networks may provide the precise temporal structure necessary for ensembles of neurons to perform specific functions, including sensory binding and memory formation. PMID- 7488854 TI - Correlated firing in sensory-motor systems. AB - Traditionally, synchronous firing of neurons has been considered to be an epiphenomenon of neuronal networks, reflecting particular properties of circuitry, but having no functional relevance. In the past few years, an alternative view has been advocated, which suggests that temporal correlations serve a role in information processing by expressing relations among the responses of distributed neurons. This hypothesis has received experimental support from recent in vivo studies performed on the sensory systems of a variety of species. These results support earlier proposals that correlated activity might have an important function in sensory-motor integration and memory. PMID- 7488856 TI - Sensory systems. PMID- 7488855 TI - Binding in models of perception and brain function. AB - The development of the concept of feature binding as fundamental to neural dynamics has made possible recent advances in the modeling of difficult problems of perception and brain function. Major weaknesses of past neural modeling (most prominently its inability to work with natural stimuli and its 'learning-time' barrier) have been traced back to improper treatment of the binding issue. Signal synchrony is now seen as playing a major role in binding. Inclusion of temporal binding in neural models has led to recent breakthroughs in solving important perceptual problems. Among them is perceptual segmentation, invariant object recognition and natural language parsing, as well as overcoming the 'learning time' barrier. PMID- 7488858 TI - p53-dependent activation of the mouse MCK gene promoter: identification of a novel p53-responsive sequence and evidence for cooperation between distinct p53 binding sites. AB - Transcriptional activation by p53 is dependent on the presence of a specific p53 binding site within control sequences of the target gene. One such target gene is the mouse muscle-specific creatine kinase (MCK) gene, which contains a p53 binding site between promoter residues -3182 and -3133 relative to the transcription start site. This DNA sequence is reported to be sufficient to confer p53-dependent activation on the MCK promoter. In contrast to this finding, evidence from promoter deletion studies suggests that sequences in the MCK promoter other than this p53 binding site also permit p53-dependent activation. To investigate this possibility, we have further examined sequences in the MCK promoter required for transcriptional activation by mouse p53. We report here identification of a second p53-responsive sequence within the MCK promoter. This novel sequence is situated between residues -177 and -81, and can confer p53 dependent, position- and orientation-independent activation on a heterologous promoter. Moreover, this sequence can specifically bind mouse and human p53. By promoter deletion studies, we provide evidence that these two elements cooperate to provide high-level, p53-dependent activation of the MCK promoter. PMID- 7488859 TI - Recessive mutations in the second largest subunit of TFIIIC suggest a new step in RNA polymerase III transcription. AB - An analysis of mutant S. cerevisiae strains selected for their ability to increase transcription by RNA polymerase (pol) III has identified 14 isolates in which this phenotype is recessive. Genetic linkage and complementation studies suggest that all 14 isolates contain recessive alleles of PCF1. PCF1 encodes the 131-kDa subunit of transcription factor IIIC (TFIIIC131) and was identified previously by dominant mutations that also increased transcription by pol III. The recessive mutation, pcf1-3, results in a conservative substitution (R728-->K) towards the carboxyl-terminus of the protein. This position is distinct from the site of the dominant mutation PCF1-1 (H190-->Y), which maps to a tetratricopeptide repeat (TPR). Site-directed mutagenesis at amino acid 728 generated one allele, pcf1-4, with a stronger phenotype than pcf1-3. Extracts from pcf1-3 and pcf1-4 strains increase pol III transcription two- to threefold and ninefold, respectively, over wild-type under conditions that permit either single or multiple rounds of initiation. The entire effect of these mutations in vitro can be accounted for by an increase in the amount of transcriptionally active TFIIIB. In contrast, PCF1-1 primarily affects the rate of preinitiation complex assembly. The genetic, molecular, and biochemical data suggest that amino acid 728 in TFIIIC131 constitutes part of a structural domain in this protein that affects TFIIIB activity by influencing a previously undefined step in transcription. This step is suggested to occur after the recruitment of TFIIIB to DNA. PMID- 7488860 TI - Identifying a transcription factor interaction site on RNA polymerase II. AB - We have generated a series of fusion proteins carrying portions of subunit IIc, the second largest subunit of Drosophila RNA polymerase I, and have used them in a domain interference assay to identify a fragment of the IIc subunit that carries the binding site for a basal transcription factor. Fusion proteins carrying a subunit IIc fragment spanning residues Ala519-Gly992 strongly inhibit promoter-driven transcription in both unfractionated nuclear extracts and in reconstituted systems. The same fusion proteins similarly inhibit dTFIIF stimulation of Pol II elongation on dC-tailed templates, suggesting that the IIc(A519-G992) fragment, which carries conserved regions D-H, interferes with transcription by binding to dTFIIF. Finally, dTFIIF can be specifically cross linked to a GST-IIc(A519-G992) fusion protein or to subunit IIc in intact Pol II. PMID- 7488861 TI - The pathway to maturity: processing of ribosomal RNA in Saccharomyces cerevisiae. AB - The 17-18S, 5.8S, and 25-28S rRNA species of eukaryotic cells are transcribed by RNA polymerase I into a single precursor molecule, from which external and internal spacer sequences are subsequently removed in an order series of nucleolytic reactions. Whereas the order of the cleavage reactions has long been established, only recently has significant progress been made in detailing the cis-acting elements and the trans-acting factors involved in this process. The use of recently developed systems for in vivo mutational analysis of yeast rDNA has greatly enhanced our knowledge of cis-acting structural features within the pre-rRNA, which are critical for correct and efficient removal of the spacer sequences. The same systems also allow a link to be forged between trans-acting processing factors and these cis-acting elements. In this review the newly obtained information will be summarized, focused predominantly on pre-rRNA processing in the yeast Saccharomyces cerevisiae. PMID- 7488857 TI - Signal transduction and activation of gene transcription by interferons. AB - Advances in the field of interferon research have identified a signal transduction pathway that initiates at a cell surface receptor and culminates at target genes in the nucleus. The binding of interferon to a transmembrane receptor stimulates the concomitant activation of tyrosine kinases of the Janus kinase (JAK) family. Subsequently, latent cytoplasmic transcription factors are activated by tyrosine phosphorylation and function as signal transducers and activators of transcription (STATs). The STATs form homomeric or heteromeric protein complexes that translocate to the nucleus to bind to specific DNA sequences in the promoters of stimulated genes. The discovery of this regulated pathway in the interferon system served as a paradigm for receptor to nucleus signal transmission by a variety of cytokines. PMID- 7488862 TI - Treatment with propylthiouracil before radioactive iodine therapy is associated with a higher treatment failure rate than therapy with radioactive iodine alone in Graves' disease. AB - Older reports have suggested that the use of antithyroid drugs with radioactive iodine-131 (RAI) results in higher rates of persistent hyperthyroidism than treatment with RAI alone. Our objective was to determine if propylthiouracil (PTU) given prior to RAI would be associated with a higher single dose RAI failure rate than treatment with RAI alone. Patients were considered treatment failures if a second dose of RAI was required to produce euthyroidism or hypothyroidism. All study patients stopped PTU at least 4 days before RAI therapy and did not receive PTU after RAI. The overall failure rate of one course of treatment in the 86 study patients was 17% (15/86). Persistent hyperthyroidism was seen in 4% of patients (2/48) treated with only RAI and in 34% of patients (13/38) receiving RAI after pretreatment with PTU (p = 0.003). Patients were treated with PTU for a mean of 151 +/- 32 days. There were no significant differences in race, gender, thyroid size, RAI dose, or days of follow-up between patients receiving RAI alone and those receiving PTU before RAI therapy. These data suggest that pretreatment with PTU leads to a higher failure rate even if PTU is discontinued at least 4 days before RAI therapy and not restarted after RAI dosing. Consideration should be given to increasing the dose of RAI in patients pretreated with PTU to ensure adequate treatment of Graves' disease. PMID- 7488863 TI - Delayed intestinal absorption of levothyroxine. AB - We report four female patients with nodular goiter (in two of the four due to Hashimoto's thyroiditis) and one male patient with frank hypothyroidism due to Hashimoto's thyroiditis in whom TSH-suppressive or replacement L-T4 therapy failed to suppress or, respectively, normalize serum TSH. As is typical in our country, our patients took L-T4 15-20 min before a light breakfast. Gastrointestinal or other diseases and drugs known to interfere with the intestinal absorption of L-T4 were not the cause of this failure. The gastrointestinal absorption test of L-T4 (1000 micrograms) was performed in four patients; in three patients it revealed peculiar abnormalities in that (i) the absorption peak was > 70% but occurred at 4 hr vs an average of 2 hr in 12 euthyroid controls (EC) and 3 hr in the 10 primary hypothyroid controls (HC); (ii) 50% of the maximal absorption occurred at 110 min vs 45 min in EC and 50 min in HC; (iii) the maximal increment in T4 absorption was between 90 and 120 min (+111%) vs between 30 and 60 min in EC (+312%) and HC (+354%). In sum, only the first part of the absorption curve of T4 was shifted to the right (in three of the four women) and this shift was more pronounced and extended to the second part of the curve in the fourth patient; in this last patient absorption peak was 44% at 180 min. Based on these results, we obtained full suppression or normalization of TSH by postponing breakfast for at least 60 min after T4 ingestion.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7488864 TI - Normal function in vivo of a homozygotic polymorphism in the human thyrotropin receptor. AB - We have demonstrated previously and association between a polymorphism in the human thyrotropin receptor gene and an increased prevalence of autoimmune thyroid disease in individuals bearing this polymorphic allele. The polymorphism involves the nucleotide base substitution of a cytosine for the wild-type adenine at the first position of codon 52 and is found generally in the heterozygotic state. Such change results in the substitution of a threonine for the wild-type proline at this position in the receptor protein sequence. The resulting protein would lack a beta turn (at position 52) in a potential loop conformation, and thus would have a significantly altered three-dimensional conformation. The biologic consequences of this conformational change in the receptor are unknown, but may involve altered function or immunogenicity. We report here two individuals with normal thyroid function who are homozygous for the thyrotropin receptor polymorphism, suggesting that the altered receptor is able to respond normally to thyrotropin with respect to the maintenance of the euthyroid state. PMID- 7488865 TI - Establishment and characterization of an antihuman thyrotropin (TSH) receptor specific CD4+ T cell line from a patient with Graves' disease: evidence for multiple T cell epitopes on the TSH receptor including the transmembrane domain. AB - From the peripheral lymphocytes of a patient with Graves' disease, we established a T cell line using its reaction to a pool of 49 synthetic peptides corresponding to the entire human thyrotropin receptor (TSHR) sequence. This T cell line showed a specific response to the pool of peptides in a microproliferation assay (stimulation index: 4.8). Flow cytometry analysis revealed that the cell surface markers were CD4+ CD8-, T cell receptor (TcR) alpha beta+, and Tcr gamma delta-. To investigate T cell epitopes on TSHR, the T cell line reacted well against three groups: the N-terminal (amino acids 31-169) and C-terminal (338-420) regions of the extracellular domain and the N-terminal half (441-661) of the transmembrane domain of the receptor. This suggests a multiplicity of T cell epitopes on the TSHR, and was further supported by analysis of TcR gene expression in the cell line that showed the expression of 5 V alpha genes; V alpha-1, 2, 10, 20, and w25. In conclusion, the results of the present study indicated multiple T cell epitopes on the TSHR molecule including the transmembrane domain. PMID- 7488866 TI - Effects of monoclonal antibody against CD45RB on peripheral blood mononuclear cell proliferation and on HLA-DR and adhesion molecule expression on thyrocytes of patients with autoimmune thyroid disease. AB - To evaluate the role of CD45 (especially that of the ectodomain region B) on immunocyte-thyrocyte signaling in patients with autoimmune thyroid disease (AITD), we have examined the in vitro and in vivo effects of a monoclonal antibody (mAb) against with CD45RB, termed MT3. MT3 was added to cultured peripheral blood mononuclear cells (PBMC) from patients with AITD and was additionally injected into severe combined immunodeficient (SCID) mice to which Graves' thyroid cells and intrathyroidal lymphocytes were engrafted. MT3 stimulated proliferation of PBMC when cultured for 2 to 3 days in patients with Hashimoto's thyroiditis (HT) and Graves' disease (GD) and in normal controls (NC). However, when cultured for 7 days, the stimulation index [SI: counts per minute (cpm) with mAb/cpm without mAb] was lowered by MT3 in NC and GD patients. However, the mean SI was not lowered in patients with HT. In SCID mice, the concentrations of human immunoglobulin G, antithyroglobulin and antithyroperoxidase antibodies in sera were not significantly changed by injecting MT3. The expression of human leukocyte antigen (HLA)-DR and intercellular adhesion molecule (ICAM)-1 on engrafted human thyrocytes decreased after the tissues were engrafted into the control mice to which vehicle alone was injected. However, in the mice injected with MT3, HLA-DR and ICAM-1 expression remained high or up-regulated by the injection.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7488867 TI - Prevalence of incidental thyroid disease in a relatively low iodine intake area. AB - An ultrasonographic survey of thyroid abnormalities was conducted in 547 consecutive apparently normal overweight subjects (380 females and 167 males), aged 27-58 years in an urban area with relatively low iodine intake (mean daily urinary iodine excretion: 10.6 micrograms/dL). Individuals with any previous thyroid disease or familial thyroid pathology were excluded. In 240 subjects (44%) high resolution ultrasonography of the thyroid was considered normal. In 307 individuals (56%) abnormalities of the echo structure (39%) or thyroid nodular disease (17%) were detected by ultrasonography. Marked heterogeneity of the echo structure that was considered suggestive of chronic autoimmune thyroiditis was present in 81 subjects (15%). In 72 of these patients the serum anti-TPO levels were positive by a sensitive RIA. Thyroid nodules either solid or predominantly cystic were present in 90 subjects (17%). Eighteen patients had a relatively large nodule (diameter 15-18 mm). Eleven of these nodules were missed at clinical examination. Fine needle aspiration cytology was performed in 14 patients and 7 individuals underwent thyroid surgery. In 6 subjects the pathologic diagnosis was benign adenomatous goiter and one patient had a follicular carcinoma. Thyroid function studies confirmed subclinical hypothyroidism in 27 patients (4.9%), all of them with elevated serum anti-TPO autoantibodies levels. It was concluded that the overall occurrence of thyroid disease is more common than suspected by clinical examination. PMID- 7488868 TI - Incidence of hypertrophic cardiomyopathy in hypothyroidism. AB - In 13 consecutive severely hypothyroid patients no sign of endocrine cardiomyopathy in the form of asymmetric septal hypertrophy (ASH) could be disclosed by M-mode and two-dimensional (2D) echocardiography on examination prior to thyroxine replacement therapy. In previous investigations ASH was demonstrated to be almost invariably present in untreated hypothyroidism irrespective of thyrotropin levels. Consequently application of positive inotropic and afterload reducing agents that may invoke deleterious effects in ASH has been considered hazardous in hypothyroidism even when indicated by concurrent other heart disease. Determination of exact confidence limits reveals that the proportional incidence of hypothyroid hypertrophic cardiomyopathy could not exceed 24.7% with 95% probability. We conclude that ASH is not necessarily inherent in hypothyroidism. PMID- 7488869 TI - Peripheral neuropathy in subclinical hypothyroidism. AB - Alterations in peripheral nerves are well documented in overt myxedema but not in subclinical hypothyroidism. We performed electrophysiologic studies to investigate such abnormalities in patients with normal serum total T4 and hyperresponsiveness of TSH to TRH, either with normal or high levels of basal circulating TSH. Subjects were divided in three groups: (i) Hypothyroidism Stage I (group () (n = 17, mean age = 39 +/- 34 years), T4 = 9 +/- 0.7 micrograms/dL, TSH = 4.3 +/- 0.4 microU/mL, sTSH post-TRH (peak value) = 37.6 +/- 1.6 microU/mL; (ii) Hypothyroidism Stage II (group II) (n = 10, mean age: 43 +/- 6 years), T4 = 7.7 +/- 0.8 microgram/dL, TSH = 20 +/- 5 microU/mL, TSH post-TRH > 50 microU/mL; (iii) Control Group (n = 20, mean age 41 +/- 5 years), healthy subjects. All patients and controls were women. TRH test consisted in the i.v. injection of 200 micrograms TRH (normal peak value up to 25 microU/mL, normal basal TSH < 5.5 microU/mL. None of the patients had carpal tunnel syndrome or any other neurological or metabolic disturbances. We studied the distal motor latencies, motor and sensory amplitudes, and nerve conduction velocities. The motor parameters were measured in the median and external sciatic popliteal (ESP) nerves, and the sensory parameters in the median and sural nerves. In most cases values were obtained from both right and left nerves. Motor parameters: no differences were found between all groups for conduction velocities (CV).(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7488870 TI - Evaluation of somatostatin as a plasma tumor marker in medullary thyroid carcinoma. AB - Somatostatin (SRIF) immunoreactivity has been frequently reported in tumor tissues of cell types, belonging to the APUD system, including medullary thyroid carcinoma (MTC). However, the value of SRIF as a plasma tumor marker for MTC is controversial. We have measured SRIF plasma levels in 35 patients with different stages of MTC to evaluate the use of SRIF as a plasma tumor marker compared to the current "gold standard" calcitonin (CT). The median SRIF value in healthy controls was 36.5 pg/mL, the upper limit of normal was defined at the controls. The median value was 28 pg/mL (p = 0.37, Mann-Whitney U test). Five patients in the control group and three in the MTC group had SRIF levels that exceed the 95th percentile. SRIF and CT levels correlated only weakly (0.38), as determined by the Spearman rank order correlation test. Pentagastrin stimulation led to a diagnostic increase in SRIF levels in only one of five MTC patients. During selective venous catheterization, diagnostic gradients for CT, allowing tumor localization, could be demonstrated, whereas measurement of SRIF levels did not aid in tumor detection. Although SRIF immunostaining may be valuable as an additional marker in the histochemical diagnosis of MTC, SRIF has no value as a plasma tumor marker in the diagnosis of this disease. PMID- 7488871 TI - Homing of 51Cr-labeled human peripheral lymphocytes to Graves' thyroid tissue xenografted into SCID mice. AB - We have recently described a NUDE/SCID mouse model that has been useful for the study of human thyroid autoimmunity in in vivo conditions. The reappearance of lymphocytic infiltration in Graves' thyroid tissue and a humoral response in SCID mice (rexenografted with normalized thyroid tissues from NUDE mice) was detected only if autologous Graves' human peripheral lymphocytes (PBMC) were engrafted into the same animals. Therefore it was presumed that some autoreactive PBMC directed themselves to the thyroid. However, there was previously no direct evidence regarding the trafficking of the engrafted PBMC to the target tissue. To elucidate this point we have studied the migration of 51Cr-labeled PBMC in SCID mice. Human thyroid tissue from six Graves' disease (GD) patients and six patients with nontoxic nodular goiter were initially xenografted into NUDE mice for 8 weeks. The same tissues were retrieved and rexenografted into several "virgin" SCID mice, i.e., no previous xenografts. Autologous PBMC were isolated from blood of the same patients obtained at the time of the tissue rexenograftment and labeled with radioactive 51Cr. Twenty million labeled PBMC were engrafted into each SCID mouse. The distribution of labeled lymphocytes into mouse organs and trafficking into Graves' and normal xenografts was measured. A significant amount of radioactivity in Graves' xenografts was detected after 1 week with the peak of radioactivity at 2-3 weeks. This radioactivity was significantly higher than radioactivity in surrounding tissues (skin, muscle). In contrast, homing of autologous lymphocytes into normal paranodular thyroid tissue was very minimal; the radioactivity of GD thyroid xenografts with engrafted autologous lymphocytes was significantly higher than that of normal tissues.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7488872 TI - Novel thyrotropin (TSH)-TSH antibody complex in a healthy woman and her neonates. AB - A novel high-molecular-weight (MW) form of immunoreactive TSH [35,000 Da on Sephacryl S-200HR gel chromatography (S-200 chromatography)] was documented in a 32-year-old healthy woman who delivered two neonates with transient hyperthyrotropinemia. Her TSH levels ranged from 21.2 to 53.9 mU/L on different days or from 11.0 to 48.1 mU/L by the different immunoradiometric assay methods. The IgG fractions showed specific 125I-labeled hTSH binding and inhibited in vitro cAMP increase induced by hTSH but not that induced by bTSH. On protein G Superose HR affinity chromatography (protein G chromatography) equilibrated with 10 mmol/L sodium/potassium phosphate buffer (PB) followed by elution with 0.1mol/L glycine buffer, 95-99% of her TSH immunoreactivity eluted in the latter (bound) fraction while almost all was in the former (unbound) fraction in the control serum containing authentic hTSH. However, after dialysis of this bound fraction overnight with PB adding 0.5 mol/L NaCl (PB/NaCl), which exhibited greater ionic strength than PB, almost all TSH immunoreactivity changed from the bound fraction into the unbound fraction on the protein G chromatography equilibrated with PB/NaCl. These data indicate that the novel immunoreactive TSH was due to hTSH and hTSH-specific antibody complex, and dissociation of the complex may be incomplete on direct S-200. The immunoreactive TSH showed high MW form (35,000 Da). The dissociation may be almost complete during dialysis with greater ionic strength; the native TSH then appeared to be of formal size. PMID- 7488873 TI - Acute compartment syndrome in a hypothyroid patient. AB - Acute compartment syndrome (ACS) is caused by conditions that either decrease the size of the compartment of increase the content of the compartment. Intracompartmental pressures are raised, thereby initiating a sequence of events that results in myoneural injury. Although limb trauma is the most common and well-organized etiology of ACS, the cause is frequently not readily apparent. Thyroid disease is not a commonly recognized cause of ACS; only one case of ACS associated with hypothyroidism has been previously reported. We now describe a second case of ACS in a patient with severe hypothyroidism and discuss the possible pathogenesis of this association. PMID- 7488875 TI - beta-Galactoside alpha-2,3-sialyltransferase messenger RNA increases in thyrotrophs of hypothyroid mice. AB - Our goal was to learn about the cellular mechanism(s) responsible for the increased sialylation of thyroid-stimulating hormone (TSH) during hypothyroidism. We used in situ hybridization to examine the beta-galactoside alpha-2,3 sialyltransferase mRNA content in thyrotrophs and corticotrophs of euthyroid and hypothyroid [propylthiouracil (PTU)-treated] mice. The alpha-2,3 sialyltransferase mRNA level was 66, 70, 296, and 223% higher in hypothyroid thyrotrophs than in euthyroid thyrotrophs after mice received PTU for 1, 2, 3, and 6 weeks, respectively. Increased transcription of this gene, or mRNA stabilization, may partially explain the increased sialylation of TSH during hypothyroidism. PMID- 7488874 TI - Factors that enhance Escherichia coli-expressed TR beta binding to T3 and DNA. AB - Thyroid hormone receptors (TRs) recently have been produced in E. coli by several laboratories. We produced E. coli-expressed human TR beta using the histidine/fusion protein system. Surprisingly, we observed that reticulocyte lysate, nonspecific proteins, and 1% Triton X dramatically increased both the T3- and DNA-binding activities of human TR beta. These studies demonstrate that there are a number of factors that will enhance ligand and DNA binding of E. coli expressed TR beta. Addition of these factors to reaction samples containing E. coli-expressed TRs will help to optimize measurement conditions. These findings also suggest that experiments in which cellular proteins are added to highly purified TR preparations may require controls to eliminate contributions by nonspecific proteins. PMID- 7488878 TI - Involvement of the orbital fibroblast and TSH receptor in the pathogenesis of Graves' ophthalmopathy. AB - Our concepts and understanding of the etiology, evolution, and propagation of Graves' ophthalmopathy have become much more sophisticated that they were 10 years ago. Given our current state of knowledge, the following scheme for the pathogenesis of Graves' ophthalmopathy can be proposed. Circulating T cells in patients with Graves' disease, directed against an antigen on thyroid follicular cells, recognize antigenic epitopes that are shared by tissues contained in the retroorbital space. Of the cell types residing in these tissues, fibroblasts are most likely to act as both target and effector cells of the retroorbital immune process. This includes those fibroblasts present in the perimysium of extraocular muscles, which do not appear to be immunologically different from fibroblasts located in the retroorbital connective tissue. By contrast, convincing evidence implicating the human extraocular myocyte itself (rather than the tissue conglomerate of extraocular muscle) as a primary target in GO remains to be demonstrated. Together with adipocytes, fibroblasts may also serve as target and effector cells in pretibial myxedema. How autoreactive T cells escape deletion by the immune system and come to be directed against a self-antigen presented by cells residing in the thyroid gland and extrathyroidal locations is unknown. T cells are recruited to and infiltrate the orbit via certain adhesion receptors, which may also play a costimulatory role in T cell activation and facilitate antigen recognition. Analysis of variable region gene usage of the T cell antigen receptors in retroorbital T cells of patients with active GO reveals limited variability, suggesting that antigen-driven selection and/or expansion of specific T cells may occur early in the evolution of GO.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7488876 TI - Influence of nonesterified fatty acids and lysolecithins on thyroxine binding to thyroxine-binding globulin and transthyretin. AB - The hydrolysis of lecithin by phospholipase produces equimolar amounts of nonesterified fatty acids (NEFAs) and lysolecithin. In this study, we have evaluated the effect of lysolecithins and NEFAs on thyroid hormone binding by examining their interactions with thyroxine-binding globulin (TBG)(serum 1:10,000 dilution) and purified transthyretin (TTR). Unsaturated NEFAs (palmitoleic, oleic, linolenic, arachidonic, eicosapentaenoic, and docosahexaenoic acid) inhibited [125I]T4 binding to TBG. Their affinities, relative to unlabeled T4, ranged from 0.005 to 0.0016%, except for oleic acid with relative affinity of < 0.0005%. Saturated NEFAs, lauric, myristic, palmitic, and stearic acid were inactive. After purification by high-performance liquid chromatography, 1-oleoyl and 2-oleoyl lysolecithin displaced [125I]T4 from TBG with an affinity of 0.0006 and 0.0005%, respectively. On a molar basis, this affinity was approximately 10 fold lower than arachidonic acid, the most potent NEFA in inhibiting T4 binding to TBG in this assay system. Of all the NEFAs tested, only arachidonic acid inhibited [125I]T4 binding to TTR, with an affinity relative to unlabeled T4 of 0.49%. 1-Oleoyl, 1-palmitoyl, and 1-stearoyl lysolecithin were without effect on TTR binding. The T4-displacing effects of NEFAs are markedly attenuated by their extensive binding to albumin. Using purified [14C]NEFA preparations and heptane partitioning, the mean unbound percentages of linoleic, eicosapentaenoic, and docosahexaenoic acid in undiluted normal human serum were 0.00099, 0.0050, and 0.0042%, respectively (n = 3). In view of the very high degree of albumin binding of NEFAs, studies in diluted serum will grossly overestimate their competitor potency. The affinities of lysolecithins for the T4 binding sites of TBG and TTR are lower than those of NEFAs and depend on the fatty acid component. Lysolecithins are unlikely to influence plasma protein binding of T4 during critical illness. PMID- 7488877 TI - The G2M arrest caused by iodide is unrelated to the effects of iodide at adenylate cyclase. AB - Previously, we have shown that the iodide was able to inhibit TSH induced thyrocyte proliferation by arresting the cell cycle at G0G1 and G2M, suggesting that the iodide may be exerting its effects through more than the TSH-adenylate cyclase-cAMP system. To confirm the effects of iodide on the adenylate cyclase (AC) system, forskolin- and dibutyryl-cyclic-AMP (dBcAMP)-stimulated FRTL5 thyroid cells were exposed to inhibitory concentrations of iodide and the resultant effects on the cell cycle were compared to the effects observed with TSH, using flow cytometric DNA analysis. Forskolin stimulated the proliferation of FRTL5 cells in a dose-dependent manner. Cell numbers rose from baseline by 169 +/- 4% to peak at 10 microM forskolin. Interestingly, 100 microM forskolin inhibited cell proliferation, causing cell numbers to fall by approximately 50%. Iodide inhibited forskolin-induced proliferation to baseline levels. However, the pattern of cell cycle perturbation was different to that with TSH-stimulated cells. There were no differences in the proportion of cells in G0G1 between forskolin alone and forskolin + NaI, while there was a marked fall in the proportion of cells in S phase, indicating possible partial arrest at G0G1. Furthermore, there was a marked accumulation of cells in G2M over and above that found with TSH + NaI, indicating arrest at G2M. dBcAMP maximally stimulated cell numbers to rise from baseline by 125% with 1 mM dBcAMP. Again, higher concentrations of the mitogen had an inhibitory effect on proliferation. The addition of NaI inhibited dBcAMP stimulated cell proliferation. PMID- 7488879 TI - "Variations" in health. PMID- 7488880 TI - Antenatal screening for carriers of hepatitis B virus. PMID- 7488881 TI - Second opinions for patients with cancer. PMID- 7488882 TI - European Union policy and health. PMID- 7488884 TI - Tighter medical controls proposed for boxing. PMID- 7488883 TI - Computer based prescribing. PMID- 7488885 TI - Dutch doctor given suspended sentence over euthanasia case. PMID- 7488887 TI - Health chiefs review London's helicopter ambulance service. PMID- 7488888 TI - Patients given LSD may be able to claim compensation. PMID- 7488886 TI - New York attacks raccoon rabies from the air. PMID- 7488889 TI - Mandatory rural practice proposed in India. PMID- 7488890 TI - Prenatal testing leads to row in The Netherlands. PMID- 7488891 TI - Half a million French are infected with hepatitis C virus. PMID- 7488892 TI - Republicans subtly reform US medicine. PMID- 7488893 TI - UKCCCR multicentre randomised controlled trial of one and two view mammography in breast cancer screening. AB - OBJECTIVE: To compare one view (oblique) and two view (oblique and craniocaudal) mammography in breast cancer screening. DESIGN: Randomised controlled trial. SETTING: Nine breast screening centres in England. SUBJECTS: 40,163 women aged 50 64 attending their first breast screening examination. INTERVENTIONS: Women were randomised to have one view mammography, two view mammography, or two view mammography in which one view was read by one reader and both views were read by another. MAIN OUTCOME MEASURES: Prevalence of cancer detected, recall rates, cost per cancer detected, and marginal cost per extra cancer detected. RESULTS: Two view mammography detected 24% more women with breast cancer (95% confidence interval 16% to 31%) than one view mammography. Prevalence of detected cancer was 6.84 with two view mammography and 5.52 per 1000 women with one view. The proportion of women recalled for assessment was 15% lower (95% confidence interval 6% to 23%) with two view (6.97%) than with one view (8.16%) mammography. The cost of two view screening was higher (26.46 pounds compared with 22.00 pounds per examination) but the average cost per cancer detected was similar (5330 pounds compared with 5310 pounds) and the marginal cost per extra cancer detected with two views was similar to the average cost (5400 pounds). CONCLUSION: Two view mammography is medically more effective than one view; it detects more cancers and reduces recall rates; it is also similarly cost effective financially. PMID- 7488894 TI - Prospective study of endogenous sex hormones and fatal cardiovascular disease in postmenopausal women. AB - OBJECTIVES: To examine the association between androstenedione, total and bioavailable testosterone, oestrone, and total and bioavailable oestradiol concentrations and the risk of death from cardiovascular and ischaemic heart disease. DESIGN: 19 year old population based prospective study with 99.9% follow up. SETTING: Rancho Bernardo, California. SUBJECTS: 651 postmenopausal women, none taking oestrogen. MAIN OUTCOME MEASURES: Concentrations of plasma sex hormones measured by radioimmunoassay in an endocrinology research laboratory. Cardiovascular and ischaemic heart disease deaths assessed by death certificate; 85% of 30% sample validated by record review. RESULTS: Age adjusted concentrations of sex hormones did not differ significantly in women with and without a history of heart disease at baseline and did not predict cardiovascular death or death from ischaemic heart disease. Most 95% confidence intervals for the age adjusted relative risk of cardiovascular death or death from ischaemic heart disease were narrow, and all included one. Endogenous oestrogen concentrations were not associated with significantly more favourable risk factors for heart disease, and testosterone was not associated with less favourable risk factors. CONCLUSION: These prospective data do not support a causal or preventive role for endogenous oestrogens or androgens and cardiovascular mortality in older women. PMID- 7488895 TI - Proposal for routine antenatal screening at 14 weeks for hepatitis B surface antigen. Dutch Study Group on Prevention of Neonatal Hepatitis. AB - OBJECTIVE: To develop a low cost, high compliance screening programme for identification of carriers of hepatitis B surface antigen in the obstetric population of the Netherlands. DESIGN: A seven year open, descriptive study of screening for hepatitis B surface antigen as part of routine prenatal laboratory testing at 14 weeks of gestation. Compliance with programme evaluated by checking delivery records (hospitals) or registration of births in the 30 participating municipalities (rural area). SETTING: Three large city hospitals (two tertiary referral centres) and one rural area with a large number of home deliveries. SUBJECTS: 99,706 pregnant women applying for prenatal care for the first time. MAIN OUTCOME MEASURES: Proportion of pregnant women routinely screened; prevalence of hepatitis B surface antigen in large cities and rural area. RESULTS: Uptake of screening reached 97% in the hospitals after inclusion of 10% screened at delivery; the estimated uptake in the rural area was > 95%. Prevalence of hepatitis B surface antigen was 1.6% in the large cities and 0.3% in the rural area. For screening at delivery the prevalence was 2.5 times higher (4%, P < 0.01) than for screening at week 14 of gestation. CONCLUSION: Incorporation of universal testing for hepatitis B surface antigen into routine prenatal laboratory testing is practical; high compliance is achieved when screening is supplemented with rapid screening at delivery for those who escaped routine prenatal care. PMID- 7488898 TI - Angio-oedema associated with risperidone. PMID- 7488897 TI - Low weight gain in infancy and suicide in adult life. PMID- 7488896 TI - Programme for preventing perinatal hepatitis B infection through screening of pregnant women and immunisation of infants of infected mothers in The Netherlands, 1989-92. National Hepatitis B Steering Committee. AB - OBJECTIVES: To launch a programme for the prevention of perinatal infection with hepatitis B in the Netherlands. DESIGN: Routine antenatal screening and intervention programme. SETTING: Community antenatal programme, the Netherlands. SUBJECTS: Infants of mothers who were carriers of hepatitis B detected by routine screening. INTERVENTIONS: Infants of infected mothers received hepatitis B immunoglobulin at birth and four doses of hepatitis B vaccine in conjunction with routine immunisation at 3, 4, 5, and 11 months of age. MAIN OUTCOME MEASURES: Results of screening and immunisation from 1989-92. RESULTS: The coverage of screening increased from 46% in 1989 to 84% in 1992. Hepatitis B surface antigen was detected in 2145 women (0.44%). The coverage of postnatal immunoprophylaxis in 1645 neonates born to mothers who were carriers of hepatitis B was 85% (1391); in 3% (42) there was a delay in administration of immunoglobulin of over 24 hours. In 1991, 96% (537), 95% (532), 94% (525), and 87% (489) of the infants received the first, second, third, and fourth dose of vaccine, respectively. There was considerable variation in the timing of vaccination; 17% (258) of the infants received their first dose more than two weeks late. Of the 59% (583) of infants who received the fourth dose more than two weeks beyond target age, 14% (141) also received their first dose too late. CONCLUSIONS: A prevention programme for perinatal hepatitis B in an area of low prevalence, when incorporated into existing health care, is feasible and achieves satisfactory coverage rates. Intensive follow up is needed to improve adherence to the immunisation schedule. PMID- 7488899 TI - Artificial colourings and adverse reactions. PMID- 7488900 TI - High fever induced by sulphasalazine. PMID- 7488901 TI - Painful dysaesthesia with ciprofloxacin. PMID- 7488902 TI - Prospective study of trends in referral patterns in fundholding and non fundholding practices in the Oxford region, 1990-4. AB - OBJECTIVE: To compare outpatient referral patterns in fundholding and non fundholding practices before and after the NHS reforms in April 1991. DESIGN: Prospective collection of data on general practitioners' referrals to specialist outpatient clinics between June 1990 and January 1994 and detailed comparisons of three phases--October 1990 to March 1991 (phase 1), October 1991 to March 1992 (phase 2), and October 1993 to January 1994 (phase 3). SETTING: 10 first wave fundholding practices and six non-fundholding practices in the Oxford region. SUBJECTS: Patients referred to consultant out-patient clinics. RESULTS: NHS referral rates increased in fundholding practices in phase 2 and phase 3 of the study by 8.1/1000 patients a year (95% confidence interval 5.7 to 10.5), an increase of 7.5% from phase 1 (107.3/1000) to phase 3 (115.4/1000). Non fundholders' rates increased significantly, by 25.3/1000 patients (22.5-28.1), an increase of 26.6% from phase 1 (95.0/1000) to phase 3 (120.3/1000). The fundholders' referral rates to private clinics decreased by 8.8%, whereas those from non-fundholding practices increased by 12.2%. The proportion of referrals going outside district boundaries did not change significantly. Three of the four practices entering the third and fourth wave of fundholding increased their referral rates significantly in the year before becoming fundholders. CONCLUSIONS: No evidence existed that budgetary pressures caused first wave fundholders to reduce referral rates, although the method of budget allocation may have encouraged general practitioners to inflate their referral rates in the preparatory year. Despite investment in new practice based facilities, no evidence yet exists that fundholding encourages a shift away from specialist care. PMID- 7488903 TI - Urinary incontinence: long term effectiveness of nursing intervention in primary care. PMID- 7488904 TI - Recent advances. Obstetrics. PMID- 7488905 TI - Don't twist my child's head off: iatrogenic cervical dislocation. PMID- 7488906 TI - Treatment for children with brain tumours. AB - The overall survival for children with malignant brain tumours has not greatly improved in recent years, so that apparently innovative treatments may seem attractive, especially in cases with a poor prognosis. Problems associated with treating such children include balancing the need to provide optimum treatment for individual patients with the need to properly evaluate advances in treatment in clinical trials and determining who should be responsible for decisions about funding such treatments. Treatment abroad is rarely justified when judged on the basis of whether the treatment is available in Britain (or European Community) and whether there is accepted evidence for its clinical benefit. However, referral to specialist paediatric centres is essential. The range of daily experience in such centres allows an educated choice between established and new treatments. Close collaboration between these centres is essential for clinical trials. PMID- 7488908 TI - ABC of medical computing. Digital imaging. PMID- 7488907 TI - Is there a moral obligation not to infect others? AB - The emergence of HIV infection and AIDS has refocused concern on the obligations surrounding the carrying and transmission of communicable diseases. This article asks three related questions: Is there a general duty not to spread contagion? Are there special obligations not to communicate disease in the workplace? And does the mode of transmission of the disease affect the ethics of transmission and, if so, how and to what extent? There seems to be a strong prima facie obligation not to harm others by making them ill where this is avoidable, and this obligation not to communicate disease applies as much to relatively trivial diseases like the common cold as it does to HIV disease. The reasonableness of expecting people to live up to this obligation, however, depends on society reciprocating the obligation in the form of providing protection and compensation. PMID- 7488909 TI - Gambling with the nation's health? Inability to reason statistically is prime cause of lottery fever. PMID- 7488910 TI - Gambling with the nation's health? Lottery has Orwellian resonances. PMID- 7488911 TI - Gambling with the nation's health? Lottery is immoral. PMID- 7488912 TI - Gambling with the nation's health? Majority of secondary school children buy tickets. PMID- 7488913 TI - Gambling with the nation's health? Doctors should concentrate on more serious issues. PMID- 7488914 TI - On sex education at school. Articles neglect the needs of young gay men. PMID- 7488915 TI - On sex education at school. Dutch surprise at British question. PMID- 7488916 TI - On sex education at school. Early education about contraception is needed. PMID- 7488917 TI - On sex education at school. Non-didactic methods are preferable. PMID- 7488918 TI - Migraine runs in families ....... but is it inherited? PMID- 7488919 TI - Screening for intracranial aneurysms. Short natural course makes screening impracticable. PMID- 7488920 TI - Controversy over new data on oral contraceptives. Risk of delay was small. PMID- 7488921 TI - Screening for intracranial aneurysms. Paper is ambiguous about number with proven aneurysm. PMID- 7488922 TI - Controversy over new data on oral contraceptives. GPs were swamped by calls. PMID- 7488923 TI - Controversy over new data on oral contraceptives. Perception of risk is affected by presentation. PMID- 7488924 TI - Screening for diabetic retinopathy. Adequate programme would save money. PMID- 7488925 TI - Screening for diabetic retinopathy. Services are still patchy in Britain. PMID- 7488926 TI - Screening for diabetic retinopathy. GPs in south London have direct access to screening service. PMID- 7488927 TI - Screening for diabetic retinopathy. Annual retinal photography is not an option in India. PMID- 7488928 TI - Screening for diabetic retinopathy. Editorial overstated likelihood of missed retinopathy. PMID- 7488929 TI - Screening for diabetic retinopathy. Screening should be organised by GPs. PMID- 7488930 TI - Screening for diabetic retinopathy. Approaching 90% sensitivity with new techniques. PMID- 7488931 TI - The new deal revisited. Obstetric trainees are against. PMID- 7488932 TI - The new deal revisited. Survey showed most anaesthetists have no experience of shifts. PMID- 7488934 TI - Polymyalgia rheumatica and giant cell arteritis. PMID- 7488935 TI - The need for an expanded protocol for the medical examination of radiation exposed persons. PMID- 7488933 TI - Combined oral contraceptives and thromboembolism. PMID- 7488936 TI - Analysis of gene expression in small numbers of purified hemopoietic progenitor cells by RT-PCR. AB - Primitive hemopoietic stem cells represent the most probable targets for genetic alterations due to exposure to ionizing irradiation or chemical carcinogens. We have applied a two-step protocol for the purification of CD34+HLA-DR-/low hemopoietic progenitor cells from cord blood (CB). CD34+ cells were isolated by monoclonal antibody (mAb) against CD34 (My10) and immunomagnetic beads. Beads were cleaved off the CD34+ cells by enzymatic treatment with chymopapain. Due to chymopapain-resistance of epitopes recognized by the used mAbs purity control of CD34+ cells and separation into CD34+HLA-DR-/low and CD34+HLA-DR+ subsets could be performed by using flow cytometry. Two miniaturized procedures were applied to isolate poly(A)+ mRNA for the reverse transcription polymerase chain reaction (RT PCR) from small numbers of CD34+HLA-DR-/low cells. In five experiments, the mean purity of immunomagnetically isolated CD34+ cells was 93.8% +/- 3.9. Flow cytometry sorting of CD34+ cells resulted in pure CD34+HLA-DR-/low populations (purity > 98.8%; range 98.8% to 99.9%; viability > 96%) with an average yield of 2600 +/- 800 cells/5 x 10(7) low density CB cells. By RT-PCR using both poly(A)+ mRNA isolation procedures, sequences corresponding to CD34 and beta 2 microglobulin were amplified from as few as 20 cells. Furthermore, a sequence independent RT-PCR (SIP-RT-PCR) was applied to amplify the cDNA derived from five erythroblasts isolated from a burst-forming unit-erythroid (BFU-E). Upon hybridization, full-length c-fos message was detected in the SIP-RT-PCR amplified material. Our data demonstrate that gene expression can be detected at the transcriptional level in small numbers of hemopoietic progenitor cells. In addition, the SIP-RT-PCR may allow the amplification of unique mRNA species when subtractive hybridization procedures are performed. The presented data should be useful to analyze gene expression in rare subsets of radiation-exposed immature hemopoietic stem/progenitor cells. PMID- 7488937 TI - Cell cycle checkpoints and repair of ionizing radiation damage. AB - Following exposure to ionizing radiation (IR), normal cells activate a delay in any phase of the cell cycle in conjunction with DNA repair mechanisms. Cell cycle delay or arrest is a programmed response that is mutable by a variety of genetic changes. DNA repair mechanisms that are responsible for the repair of otherwise lethal IR-induced double-strand breaks (DSBs) operate in a parallel pathway. The formulation of this pathway has recently been investigated, and new information regarding several mutant cell lines that are unable to execute IR-induced DSB repair are summarized. The scid mutation and defects in Ku proteins have been characterized. Molecular readouts of the properties of IR repair have been identified, including the hyperphosphorylation of the 34 kDa subunit of replication protein A. In addition, we have identified features of the G1/S IR induced checkpoint that can be influenced by p53 status, genetic background or the levels of cell cycle proteins. A further understanding of the players in these pathways is expected to lead to the identification of molecular markers for ionizing radiation damage. Examination of the changes in these proteins may be valuable in a clinical setting for documenting radiation exposure. PMID- 7488939 TI - Marker genes for cytotoxic exposure: p53. AB - The growth suppressor p53 plays an important role in the regulation of cell proliferation, DNA repair and apoptosis. In wild-type p53 expressing cells, gamma irradiation induces an increase in the level of p53 protein and these cells exhibit a G1 growth arrest. The p53-induced G1 growth arrest is abrogated in cells expressing mutant p53, or in cells where p53 is inactivated by complex formation with cellular or viral proteins such as mdm2 or the E6 proteins of human papillomavirus (HPV) 16 or HPV18. Wild-type p53 expressing cells are radiosensitive whereas mutant p53 expressing cells are radioresistant. In some cell types, p53 mutations are observed after gamma-irradiation of cells although this observation is not consistent for all cell types. Furthermore, it is not clear whether these mutations are the direct result of irradiation or secondary effects. PMID- 7488940 TI - Utility of biological membranes as indicators for radiation exposure: alterations in membrane structure and function over time. AB - In addition to interacting with genomic DNA, ionizing radiation may directly and indirectly alter the structure and function of components of the plasma membrane of eukaryotic cells. Water radiolysis generates reactive species, including superoxide, hypochlorous acid and chloride radicals that may in turn react with biological membranes, as well as with cellular DNA. Reaction of plasma membrane lipids with molecular oxygen results in lipid peroxidation of both reconstituted membranes and biological membranes, an effect that increases with decreasing dose rate. Both ionizing radiation and ultraviolet light alter functions of membrane anchored molecules, including adhesion molecules, histocompatibility complex antigens and membrane-bound growth factors. The latter growth factors represent a repertoire of growth and differentiation signals that are expressed in a nondiffusible fashion at the cell surface, and in soluble forms appearing after cleavage of their extracellular domain. The importance of cell-cell signaling via the membrane-anchored form of growth factors is becoming increasingly recognized. Expression of membrane-bound hematopoietic cytokines by eukaryotic cells is impaired after exposure to ultraviolet light, a defect in cell-cell signaling that may lead to impaired hematopoiesis. While studies suggest that permanent changes in membrane structure and function may result from radiation-induced injury to the plasma membrane and reconstituted "pure" membranes, reversibility of these defects over time requires additional study. PMID- 7488938 TI - Spontaneous and induced apoptosis after whole body radiation exposure: experimental approaches. Observations in radio-induced thymic lymphomagenesis. AB - Radio-induced thymic lymphomagenesis is associated with alterations in the balance between thymocyte subsets and cytokinetic perturbations. The objectives of this work were to investigate whether these alterations are associated with alterations in the basic levels of thymocyte apoptosis. For this purpose, we tested DNA fragmentation by gel electrophoresis, analyzed DNA content by propidium iodide staining of ethanol fixed cells and looked for DNA strand breaks on tissue sections by in situ end labeling. We described an increase of the levels of apoptosis in cultured thymocytes during the preleukemic period, while the basic levels of apoptosis observed in situ are similar in normal and in preleukemic thymuses. We propose that after leukemogenic irradiations, there is an increase of cells wherein the cell suicide program is activated but that environmental thymic factors rescue them from apoptosis. Preleukemic cells could belong to this abnormally surviving population of cells "programmed to die," wherein additional genomic abnormalities would lead to fully neoplastic transformation. PMID- 7488942 TI - Surface markers and growth factor receptors of immature hemopoietic stem cell subsets. AB - The regenerative potential of bone marrow following exposure to relatively high doses of ionizing radiation, as well as the efficacy of hemopoietic growth factor treatment, are dependent on the residual number of hemopoietic stem cells. From studies in mice in particular, evidence has been obtained that immature hemopoietic stem cells are heterogenous with respect to repopulating capacity, with one subset being capable of short-term, transient hemopoietic reconstitution and another subset of sustained reconstitution. In rhesus monkeys, CD34+, RhLA DRdull cells were identified as the small fraction of a bone marrow cell that contains reconstituting hemopoietic stem cells. The growth factor receptor phenotype of this immature cell fraction has been determined for granulocyte macrophage colony-stimulating factor (GM-CSF), interleukin 3 (IL-3), and IL-6 as well as for kit-ligand, making c-kit an especially strong growth factor receptor marker for reconstituting stem cells. In addition, it is demonstrated that CD34+ cells appear in peripheral blood after exposure to radiation and are correlated to numbers of CD34+ cells in bone marrow. This finding suggests that circulating CD34+ cells may be used as a cellular marker with prognostic significance for both the number of residual stem cells as well as regeneration of immature hemopoietic cells in bone marrow. PMID- 7488943 TI - Cytogenetics research in radiation biology. AB - Radiation cytogenetics goes back approximately six decades and not only contributed to the earliest development of radiobiology, but continues to contribute today. Contributions on three levels are outlined here. Early contributions to radiobiological theory include the nature of dose-effect curves, dose-rate and fractionation effects, and linear energy transfer (LET) effects. Understanding of the roles of aberrations in endpoints such as cell killing, mutation and carcinogenesis have more recently contributed to unraveling mechanisms in these important radiobiological effects. Finally, the study of various details of classical radiation cytogenetics, such as half chromatid exchange or sister chromatid union, has contributed to our current understanding of cytogenetic phenomena on the molecular level. PMID- 7488941 TI - Apoptosis and the assessment of radiation injury. AB - Apoptosis is a necessary form of cell death for all multicellular organisms and is controlled by the action of genes that initiate, facilitate or inhibit the process. In addition to physiological triggers, apoptosis can follow cellular injury, such as exposure to ionizing radiation. Cell death following radiation serves as a means by which the possibility of cells surviving with potentially harmful genomic damage can be reduced. Apoptosis can be identified in tissues shortly after whole body radiation; but apoptotic cells are rapidly cleared by phagocytosis, and apoptosis is unlikely to serve as a useful long-term marker of previous radiation exposure. PMID- 7488945 TI - The role of stable chromosome aberrations as biological indicators of radiation effect: studies in patients after total body irradiation and bone marrow transplantation. AB - The so-called unstable aberrations measured primarily as dicentrics and ring chromosomes are very useful as indicators of human radiation exposure. The role of stable chromosomal aberrations as indicators of radiation effects is not yet known. Stable chromosome aberrations persist longer in radiation-exposed individuals because cells with monocentric aberrations (= stable) have a higher probability of survival after cell division than multicentric or acentric aberrations (= unstable). Aberrations were investigated in 89 patients with chronic myelogenous leukemia undergoing total body irradiation (TBI) for bone marrow transplantation (BMT). Serial follow-up investigations were performed in 31 patients by analyzing lymphocytes of the peripheral blood. Patients had TBI of 10 to 12 Gy as conditioning therapy for BMT. Stable anomalies were investigated by Giemsa-banding and karyotyping in addition to standard analysis. A very high number of cells with stable aberrations was found up to five years after TBI. Investigations of the blood showed that a considerable number of cells with stable aberrations derive from radiation-damaged and clonally expanded hemopoietic precursor cells. We conclude that stable aberrations are useful as indicators of radiation effects in human beings. Clonal stable aberrations represent cellular radiation effects conserved at the stem cell level. Stable aberrations should be analyzed not only by the use of fluorescence in situ hybridization with chromosome painting probes, but also by karyotyping. The latter is necessary to detect stem cell effects by proving the clonal expansion of chromosomal aberrations, and should be performed with the assistance of an automatic chromosome analysis system. PMID- 7488944 TI - Cytogenetic research after accidental radiation exposure. AB - Currently, chromosome aberration analysis in peripheral lymphocytes is the most sensitive method to estimate individual doses in accidental radiation exposures. The assessment of dose is particularly reliable in cases with acute, uniform, whole-body exposure or after irradiation of large parts of the body. However, the scenarios of most radiation accidents largely result in partial-body exposures or a non-uniform dose distribution. This complicates dose estimation especially in cases with protracted or fractionated exposures. Problems exist also for the dose reconstruction of radiation exposures occurring a long time before sampling. To overcome these problems, the Qdr method or the "contaminated Poisson" method can be used to determine meaningful dose estimates from data based on conventional scoring of dicentrics. Scoring of so-called stable translocations by the newly developed technique of chromosome painting should be particularly useful for estimating doses of past exposures or of dose accumulation. After incorporation of radionuclides with largely localized depositions in certain organs or tissues, realistic individual dose estimates cannot be achieved. Exemplified by incidents involving larger groups of the population such as in Chernobyl and Goiania and by single cases with serious overexposures, chromosome dosimetry is evaluated in the present article. PMID- 7488946 TI - Micronucleus determination as a means to assess radiation exposure. AB - Radiation accidents require an easy to perform and rapid determination of the radiation response of the victim(s) affected by the exposure. Micronuclei in cytochalasin B blocked lymphocytes are able to serve this purpose. Micronuclei are particularly useful when it comes to screening many exposed people after a more or less homogeneous whole body exposure with a high dose rate. There are problems, as is the case for most biological indicators, with partial body (especially when it is due to internal emitters), chronic and fractionated exposure. PMID- 7488947 TI - The single cell gel electrophoresis/comet assay: a potential tool for detecting radiation-induced DNA damage in humans. AB - This review evaluates the applicability of the single cell gel (SCG)/comet assay as a tool for the biomonitoring of individuals accidently, environmentally or occupationally exposed to ionizing radiation. This technique detects single strand DNA breaks, alkali-labile damage, incomplete excision repair sites and DNA:DNA crosslinking at the level of the individual cell. The advantages of this technique include: (a) data are collected at the level of the individual cell, providing information on the intercellular distribution of damage and repair; (b) only small numbers of cells are required (i.e., only a few thousand); (c) virtually any eukaryotic cell population can be used; and (d) the assay is relatively sensitive (detection limit of 5 cGy gamma rays in human lymphocytes), simple and cost effective. What the assay lacks is specificity for radiation induced DNA damage. However, the possibility of identifying types of DNA damage specific for ionizing radiation in selected subtypes of cells may be feasible. While additional research is required before the SCG assay can readily be applied as a standard biomonitoring tool for exposure to ionizing radiation, the data collected thus far support a conclusion that such research is clearly warranted. PMID- 7488948 TI - Dose, effect severity, and imparted energy in assessing biological effects. AB - Because of the widespread efforts in cancer radioepidemiological studies to attach a value of absorbed dose to each exposed individual, the notion seems to have become prevalent that dose plays an essential role in the medical determination of the diagnosis and prognosis of the individual. This view is enhanced by the fact that, while the present quantities and units for radiological physics were developed in the context of the acute effects of large exposures to radiation, e.g., in radiotherapy where they still apply well, these same quantities and units have been used, without modification, to apply to cancer radioepidemiology in the context of low level irradiation. A principle purpose of the present communication is to show that, in medicine, dose plays a limited role even in the deterministic application of therapeutic agents, and that diagnosis and estimates of prognosis in medicine are based, not on dose, but on the severity of effect on, or damage to the organ or organs involved in a particular medical condition. Thus it is "going backward" to view estimates of the severity of effect, e.g., the fraction of cells with abnormalities, or killed, as a "biological dosimeter," rather than as a quantitative estimate of the severity of effect. The use of biological indicators is of maximum value in noncancerous disease or injury in which the severity of an effect causative for organ failure and a consequent quantal, e.g., a lethal response in the individual, can be measured with increasing accuracy by modern medical techniques.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7488949 TI - DNA-damage detection in man after radiation exposure--the comet assay--its possible application for human biomonitoring. AB - The exposure of human beings to ionizing radiation is still of great concern to occupational and environmental medicine. The goal of this workshop is to identify a panel of biological markers that could be used in humans after exposure to ionizing radiation. The comet assay or single cell gel (SCG) assay is a new method that allows efficient determination of single-strand breaks (SSB) and double-strand breaks (DSB), as well as alkali-labile sites in the DNA of single cells. In order to demonstrate the practicability of the comet assay for the detection of DNA damage caused by low doses of ionizing radiation, we exposed human peripheral blood cells to radiation in vitro. The extent of DNA damage in blood cells irradiated with x-rays (0.05-1 Gy) was significantly increased above the control values even at 0.05 Gy and shows a clear dose-relationship. To investigate the repair kinetics for x-ray-induced DNA damage following acute and chronic (fractionated) irradiation, we exposed peripheral blood to 1 Gy and examined the tail moment at different time intervals. The effect of one acute dose is repaired within two h, whereas the effect of fractionated irradiation gives a totally different result. The tail moment of the initial damage increased indicating an accumulation of the damage, and the repair activity clearly decreased. Until now, there was no data available concerning DNA damage in vivo. For this reason, we explored patients subjected to radioiodine therapy as well as a Chernobyl liquidator.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7488950 TI - Application of the premature chromosome condensation assay in simulated partial body radiation exposures: evaluation of the use of an automated metaphase-finder. AB - The premature chromosome condensation (PCC) assay has been proposed as a useful and rapid end point for biological dosimetry following accidental high-dose radiation overexposures. A major benefit of the PCC assay is that it does not require cells to divide for evaluation of cytogenetic damage. The PCC assay was performed on isolated human peripheral lymphocytes exposed in vitro to doses from 1 to 9 Gy of 250 kVp x-rays. The dose-response relationships of the frequency distribution and the yield of PCC fragments in cells were determined after one day of repair at 37 degrees C. A Qpcc approach, which involves the analysis of the yield of excess PCC fragments in damaged cells, was used to establish a dose response calibration curve. This method is identical in concept to the Qdr technique introduced by Sasaki for partial-body exposure dose-estimates using asymmetrical chromosome aberrations (i.e., dicentrics and rings) in metaphase spreads of human lymphocytes. A simulated in vitro test of a partial-body exposure to a 6-Gy dose was performed. The results from this test provided dose estimates of 5.3 +/- 0.6, 4.7 +/- 0.6, 5.0 +/- 0.6 and 4.7 +/- 0.8 Gy for the 20, 30, 50 and 75 percent component of 6-Gy irradiated cells, respectively. An automated metaphase-finding system was evaluated for use with the PCC assay. This system helped to locate PCC spreads among the mitotic inducer Chinese hamster ovary (CHO) metaphase spreads, thereby facilitating rapid scoring of samples.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7488951 TI - Biomarkers for low-level exposure causing epigenetic responses in stem cells. AB - Our current understanding of experimental in vitro and in vivo studies, as well as of epidemiological data, suggests that carcinogenesis is the result of many endogenous and exogenous factors interacting during a multi-step, multi-mechanism process. No single factor "causes" cancer. Carcinogenesis is known to involve both mutagenic and nonmutagenic processes. Therefore, the objective of this review is to examine how low dose exposure to ionizing radiation might contribute to the carcinogenic process and if measurable "biomarkers" might be used to monitor the susceptibility, exposure, biological consequence and clinical disease patterns attributable to ionizing radiation. While the primary biomarker for ionizing radiation has been DNA damage and genetic/chromosomal mutations, possible effects on apoptosis and epigenetic processes have been examined. The search for biomarkers of cytotoxic (apoptotic) and epigenetic events induced by low-level ionizing radiation was thought to be difficult in view of the fact that controlled apoptotic and epigenetic events occur constantly in a healthy body exposed to background radiation. In addition, if the stem cell is the target cell for low-level radiation exposure, detection of biomarkers in this small subpopulation of cells in any tissue places severe limitations on any practical noninvasive means to identify such biomarkers. Last, a critical re-examination of the cancer incidence data of the Hiroshima/Nagasaki A-bomb survivors is suggested in view of modifying factors such as caloric restriction and post-irradiation trauma/treatment that could affect the multi-stage nature of carcinogenesis. PMID- 7488952 TI - Pathophysiological aspects of tumor development. AB - Neoplastic transformation is one possible consequence of genomically disturbed intracellular feedback mechanisms normally governing life, differentiation, function and death of an individual cell. Neoplastic growth can be thought of as the abnormal activation of the mitotic program and/or the inactivation of programs for growth-inhibition and apoptosis. This article reviews the current knowledge on three types, or families, of proteins that act on different levels of subcellular organization and are involved in controlling the integrity of the genome, survival and death: i) the DNA-binding nuclear protein p53 inducing cell cycle arrest and apoptosis, ii) the bcl-2 family of proteins acting as regulators of prolonged survival and programmed cell death and iii) APO-1/Fas, a cell surface receptor transducing an apoptotic signal delivered either by the cell itself (cis death) or by another cell (trans death). Although much is still unknown, especially concerning the functional linkages of these three principles, the data available allow a fascinating insight into the society of cells, which we are, after all. PMID- 7488953 TI - Initiation of non-neoplastic late effects: the role of endothelium and connective tissue. AB - While early radiation lesions might be a direct consequence of parenchymal cell loss, late-radiation injury most probably develops as a consequence of functional perturbations that may involve both parenchymal and nonparenchymal elements. Damage to blood vessels and consequent perturbations in blood flow and endothelial physiology play an important role in the development of late effects. The development of late-radiation damage has been studied in three different tissue systems: the skin, kidney and central nervous system. The results suggested that damage to vascular tissue played a major role in the development of radiation-induced late effects. PMID- 7488954 TI - Heterogeneity of radiation sensitivity of hemopoietic stem cell subsets. AB - Immature hemopoietic cells are heterogeneous with respect to repopulating capacity following exposure to ionizing radiation, growth factor receptor and surface marker phenotype as well as cycling state. In the present paper, the evidence from studies in subhuman primates and mice is reviewed, which demonstrates that the heterogeneity of immature stem cells is also reflected by a differential sensitivity to radiation, the most immature stem cells with long term repopulating capacity being less sensitive to radiation than assumed on the basis of 50% lethal dose (LD50) data, and the radiation sensitivity of immature hemopoietic cells measured by the spleen-colony test. In addition, these immature cells appear to have a considerable repair capacity. These findings have a direct bearing on future strategies for the treatment of patients after accidental exposure to ionizing radiation, either by transplantation of (subsets of) stem cells or by administration of a specific combination of hemopoietic growth factors. PMID- 7488955 TI - Response of hemopoiesis in dogs to continuous low dose rate total body irradiation. AB - Among the cytotoxic agents which particularly cause damage to cell renewal systems, ionizing radiation is one of the most effective ones since it leads to inactivation of all types of proliferating cells including resting stem cells. It is the aim of this paper to present the effects of continuous low dose rate total body irradiation (TBI) on hemopoiesis in dogs. The animals were exposed to gamma rays from a 60-Co source, receiving a daily radiation dose of 0.0188 Gy for indefinite times. Sequential hematological studies performed included determinations of peripheral blood cell counts and of total cell numbers in standardized bone marrow samples, assessments of progenitor cells GM-CFC in the blood and bone marrow, and of colony-stimulating activity (CSA) in the serum. The lymphocytes, the thrombocytes and neutrophilic granulocytes uniformly showed early decreases within the first 200 to 500 days corresponding to cumulative radiation doses in the range up to 3.8 to 9 Gy, but remained stable at subnormal levels in the period up to 1,700 days of exposure. The GM-CFC numbers in bone marrow samples from the rib clearly showed a strong decrease within the first 150 days of exposure preceding the changes in the blood granulocyte concentration. A transient partial recovery of the GM-CFC was observed at later times between 700 and 1,200 days of exposure, followed by another decrease to extremely low values at cumulative doses in the range of 32 Gy.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7488956 TI - Physiological and pathophysiological aspects of the immune system contributing to a biomathematical model of lymphocytes. AB - The effects of chronic low-dose irradiation on the immune system and the lymphocytes are largely unknown. The uranium miners in the former German Democratic Republic (GDR) were exposed mainly to a local low-dose irradiation in the lung by radon and its progeny, but also to some whole-body gamma irradiation. The local irradiation led to an increased rate of lung cancer and perhaps to some increase in extrapulmonary neoplasms. But little is known about the effects on the lymphocytes circulating and recirculating to the lung. As a prerequisite for the establishment of a biomathematical model to estimate lymphocyte fluxes, and to assess the radiation effects on the lymphocytic (and stem cell) populations passing through the lung, it was necessary to establish the current knowledge with respect to the physiology and pathophysiology of the lymphocytic cell renewal systems. The data concerning lymphopoiesis and lymphocyte kinetics, which are important for the development of this model, are summarized. The distribution of lymphocytes between different compartments including the lung is taken into consideration, as well as the effects of acute and chronic irradiation on the immune system. The extracorporeal irradiation of the blood (ECIB) may serve as a model of irradiation of blood in the lung. This review shows that many data necessary for development of a detailed biomathematical model are still missing, especially data concerning details on lymphocyte production rates of their different subsets and regulatory mechanisms of the lymphocytic system. PMID- 7488958 TI - A biomathematical model of granulocytopoiesis for estimation of stem cell numbers. AB - In this paper a dynamic model of granulocytopoiesis is used to estimate stem cell numbers after acute and chronic irradiation. Regeneration data from irradiation accidents and bone marrow transplantation are investigated in the case of acute irradiation. Data from dogs living in a low-dose irradiation field are investigated for the case of chronic irradiation. The effects of acute irradiation and bone marrow transplantation are interpreted as a change of the initial conditions of the model, while the effect of chronic irradiation is interpreted as a continuous cell loss in the proliferating cell compartments of the model. The unknown initial conditions of the model are computed on the basis of real patient and dog data with the help of a suitably constructed optimization problem. The numerical solution of this optimization problem will only be sketched in this paper. In all cases discussed in this paper, a very good agreement between the model and the real data is achieved. PMID- 7488959 TI - An approach to a biomathematical model of lymphocytopoiesis. AB - Fundamental principles for the development of a biomathematical model of lymphocytopoiesis are presented in this paper. The first step in this modeling approach is the definition of appropriate anatomical compartments in order to identify dominant locations of lymphocytes in the human body, and the definition of functional compartments in order to model different maturation stages. In the second step these compartment structures are combined, and thus form the basis of a dynamical model consisting of linear differential equations. Cell balance equations are used to derive the biomathematical dynamical model which is presented using the tools of modern systems theory. As a result of intensive discussions between engineers and medical doctors, our model of lymphocytopoiesis consists of six anatomical and six functional compartments. Virtual marking technique plays a key role for the interpretation of the simulation results which are in solid agreement with biological observations. Future work is directed towards implementation of radiation damages in order to meet the final goal, namely, the evaluation of our model against the data derived from a group of chronically irradiated uranium miners. PMID- 7488957 TI - The delayed respiratory consequences of inhaled radionuclides in population exposed to nuclear catastrophe. AB - Cytological analysis of bronchoalveolar lavage (BAL) and x-ray energy dispersive spectrometry of intercellular compounds were evaluated in 19 workers ("liquidators") commissioned to clean and contain the nuclear waste after the Chernobyl catastrophe in comparison with ten control subjects. Results showed a significant increase of neutrophils, small monocyte-like alveolar macrophages (AM) and AM with large black cytoplasmic inclusions. In addition, such mineral elements as Zr, Ce, Pr, Sm, Pa, PU and Cs were detected in AM phagosomes in liquidators. These elements were reported as markers of Chernobyl-type nuclear fuel. It is recognized that BAL gives access to particulate matter present in lung parenchyma. Thus, spectrometric analysis of AM inclusions can provide reliable information about the exposure to inhaled radionuclides seven years after the catastrophe. PMID- 7488960 TI - The concept of biological markers in the field of risk assessment. AB - Using biological markers as a means to advance toxicological science has come under increasing scrutiny. The concept of using biological fluids to identify an event affecting the organism is at least as old as associating a disease with a change in the color of the urine. What is new is the perception that rapid advances, in both conceptual understanding of toxicology and in the power of analytical biology and chemistry, greatly increase the likelihood of making advances in the development and validation of biological markers. These markers are of significance to human health and the environment, as well as to risk assessment and to other techniques applicable to regulatory decision making. Biological markers can be divided into markers of exposure, effect and susceptibility. There is a continuum between exposure and effect, with some markers being more reflective of either end of the continuum. An ideal marker is one that reflects both exposure and effect. Development of markers requires an understanding of the mechanisms by which agents enter the body, distribute, undergo metabolism and elimination, interact with target cells and molecules and produce adverse effects. The human genome project is likely to lead to a further increase in the availability of susceptible markers. There are many ethical issues concerning biological markers in general and susceptibility markers in particular. Validation of biological markers is a painstaking but necessary effort that requires a concerted effort among laboratories and research organizations. PMID- 7488962 TI - Are there 'common denominators' in different radiation exposure scenarios as a target for predictive assessment? AB - The first panel discussion of the International Searle-Foundation Workshop on Biological Indicators of Exposure to Ionizing Radiation was directed at identifying 'common denominators' for the development of somatic late effects. The presented accidents were analyzed in terms of occurrence of total body or partial body irradiation, external or internal contamination, onset of early signs or symptoms of acute radiation syndrome, reduction of life expectancy, and occurrence of neoplastic or non-neoplastic disorders. Evidence of radiation effects was sought in the victims' cell systems and in any cellular or molecular response to ionizing radiation, and whether other complicating factors could be established. From the results presented during the meeting, as well as from a review of the relevant literature, it is, however, rather unlikely that a single parameter will emerge by which the late effects of ionizing radiation in man can be determined easily. Instead, a couple of acute and chronic effects of exposure to ionizing radiation could be identified which are more or less common to any given exposure scenario. PMID- 7488961 TI - Modeling of radiogenic responses induced by fractionated irradiation in malignant and normal tissue. AB - The aim of this contribution is to outline how methods of system analysis, control theory and computer science can be applied to simulate malignant and normal cell growth and to optimize cancer treatment. Based on biological observations and cell kinetic data, our group has constructed three types of computer models: 1) A cell cycle model describing the spatial (3D) and temporal growth of tumor spheroids; 2) A compartment model describing the growth of rapidly proliferating normal cells; 3) A compartment model simulating slowly proliferating normal tissues. These growth models have been extended by an irradiation model based on the linear-quadratic survival function. Different clinical fractionation schemes (standard-, super-, hyperfractionation and weekly high single dose) have been applied to the tissues mentioned above. The simulation results show that in the case of irradiating a rapidly growing tumor spheroid the hyperfractionation (3 x 1-1.5 Gy per day) leads to a particularly good anti-tumor effectiveness. On the other hand, the radiogenic response of rapidly growing normal tissue to a hyperfractionated treatment schedule is severe. The same result is observed when simulating the late reaction on slowly growing parenchymal tissue. Therefore, this therapeutic modality is ensured only if the overall dose is reduced from DTOTAL = 60 Gy to DTOTAL = 50 Gy.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7488963 TI - What is the status of molecular and cell biology approaches relevant to radiation effects assessment? PMID- 7488964 TI - The use of biological indicators to assess consequences of radiation exposure at the cell system level. PMID- 7488965 TI - Biologic indicators of exposure: are markers associated with oncogenesis useful as biologic markers of effect? AB - Radiation-induced molecular and cellular alterations play an important role in the transformation of a normal cell into a cancer cell. However, the basic molecular and cellular alterations upon exposure to ionizing irradiation are still poorly understood. Identification of such alterations would be of importance for the assessment of exposure dose, as well as for the assessment of an exposed individual's risk of developing cancer. Extensive studies of the mechanisms of oncogenesis have led to the identification of altered genes, such as proto-oncogenes and tumor suppressor genes as well as other genes intimately involved in cellular proliferation and differentiation, that are more or less frequently associated with a variety of human malignancies. It can be assumed that at least some of these mechanisms are associated with radiation-induced oncogenesis. The longevity of stem cells, particularly those of the hemopoietic system, makes them the prime target cell population to accumulate genetic alterations due to exposure to a variety of agents. Improvements in purification strategies for hemopoietic stem cells, as well as the availability of sensitive techniques such as the polymerase chain reaction (PCR) and flow cytometry analysis, should allow in-depth studies at the molecular and cellular level after exposure to physical and chemical agents. PMID- 7488966 TI - International program on the health effects of the Chernobyl accident. AB - The International Program on the Health Effects of the Chernobyl Accident (IPHECA) was established by the World Health Organization (WHO) in 1991. Currently, the technical part of IPHECA consists of five projects addressing the following areas of priority health problems or needs: thyroid, hematology, brain damage in utero, epidemiological registry and oral health. Important findings are: 1) a significant increase of thyroid cancer in children in Belarus and Ukraine since 1989, and in Russia since 1992 though not so pronounced. A relationship between detected thyroid cancers and radiation exposure is yet to be established, 2) no increase yet in the incidence of hemoblastoses in the three States, 3) no relationship established between mental retardation and radiation exposure in utero in 4,500 children investigated. The importance of dosimetry and biological indicators of radiation damage has been recognized by IPHECA. Several methods of biological and physical dosimetry are being employed using instrumentation provided by IPHECA. Some preliminary results indicate: 1) unstable aberrations can indicate an integral exposure but it is heavily biased to recent exposures, 2) when comparing healthy persons and patients with hematological diseases in contaminated areas, there is a higher ratio of total aberrations compared to their background and that the level of stable is lower than unstable aberrations, and 3) by applying electron spin resonance (ESR) it has been shown that the individual distribution of doses approaches a log-normal one, especially for adults, and that a peak shift towards higher doses is noticeable for children. PMID- 7488967 TI - Cancer risks and biomarker studies in the atomic bomb survivors. AB - This paper describes recent studies on cancer incidence among Japanese atomic bomb survivors. The report includes a brief description of the primary atomic bomb survivor cohorts. The results illustrate the magnitude of the excess risks and show how these risks vary with sex, age at exposure and time. Descriptions are presented in terms of both excess relative risks and excess absolute rates. The paper also includes a summary of the current status of research on biomarkers of radiation exposure in the atomic bomb survivors. PMID- 7488968 TI - Medical effects of exposure of human beings to fallout radiation from a thermonuclear explosion. AB - On March 1, 1954, after detonation of a thermonuclear device on Bikini atoll, an unexpected wind shift resulted in the deposition of radioactive fallout on inhabited atolls. The fallout radiation caused fleeting systemic effects, dose dependent depression of hematopoiesis and skin burns primarily due to the beta ray component of the fission radionuclides. Within a few weeks, hematopoietic recovery was substantial but slight depression of blood counts was maintained for several years. One case of fatal acute myeloblastic leukemia developed in a boy receiving 1.9 Gy as an infant. Cretinism developed in two boys exposed as infants with estimated thyroidal dose in excess of 50 Gy. Chemical hypothyroidism was detected in several persons. Thyroid adenomas and cancer commenced appearance ten years after exposure and became a major long-term medical problem. There have been no late effects attributable to the beta burns 40 years after exposure. Internal contamination from ingestion and inhalation of radionuclides is detectable. The doses are comparable to background levels in the U.S. There is no detectible decrease in longevity of the exposed Marshallese compared to an unexposed Marshallese population. PMID- 7488969 TI - Health effects of radiation incidents in the southern Urals. AB - This article discusses the most important information on health effects in the Urals region (Russia) of residents exposed to radiation from activities of a weapon plutonium separation plant. The population residing on the contaminated territory was exposed to chronic combined irradiation (external gamma-irradiation and internal irradiation due to Sr-90 and Cs-137). The red bone marrow (RBM) was the critical organ affected as a result of radiation events in the Urals. In the early period, after the discharges of radioactive wastes into the river Techa (about 3 M Ci) started, cases of chronic radiation sickness (CRS; 940 cases, in total), postirradiation reactions manifested by changes in blood parameters (e.g., leukopenia, thrombocytopenia, granulocytopenia), nervous system disorders, immunity changes and ostealgic syndrome were registered in a portion of those riverside village residents who had received the highest doses. Increased leukemia and cancer mortality and morbidity rates were noted among this population in later periods. No late effects were observed in residents exposed to an explosion in a radioactive waste depot in September, 1957 when radioactive wastes with about 20 M Ci of activity were released into the environment. Similarly, the offspring of the residents exposed on the Techa also did not display any late effects. The data about the possibilities of long-term (43-45 years after the start of exposure) biological indication of chronic internal exposure are presented. The methods used in the study include in situ fluorescent hybridization, analysis of mutations in the TCR gene of peripheral blood lymphocytes and erythrocyte mutations in the glycophorine A system. No dependence of genomic translocations and mutations in glycophorine A on cumulative exposure dose to RBM was traced. PMID- 7488970 TI - Chernobyl experience: biological indicators of exposure to ionizing radiation. AB - Using the Chernobyl accident as an example, an attempt is made to consider the possibility of using the biological markers of exposure and effects of exposure to ionizing radiation in relation to biology dosimetry, and to predict early and late nonstochastic and stochastic radiation consequences. The biological dosimetry was based on the three markers: chromosome aberrations of peripheral blood lymphocytes, dynamics of blood cell (lymphocytes, neutrophils) counts and electron spin resonance (ESR) of tooth enamel. The first two methods can be applied in a short period of time (days or weeks) after exposure and only after high doses (> 0.5-1 Gy) of acute total body irradiation (TBI). The ESR tooth enamel method possesses dosimetric value at all conditions of uniform gamma TBI (acute, prolonged, chronic and high as well as low level of doses) and at any time after exposure. The low limit of sensitivity of the ESR test is about 0.1 Gy. The use of biological markers of effects of radiation exposure as early diagnostic signs was limited to clinical significant disorders of hemopoietic, immune systems and skin in conditions of acute high-dose irradiation. In cases of acute or prolonged irradiation in low doses, many changes on the cellular as well as organism level were discovered. However, there were not enough data on radiation specificity or dose dependence of these changes. Hence they cannot be considered as the indicators of clinically significant early and late nonstochastic effects. The role of biological markers of stochastic effects in clinical practice is discussed herein. PMID- 7488971 TI - Cellular mechanisms of protection and repair induced by radiation exposure and their consequences for cell system responses. AB - The complex biological systems that constitute living organisms operate at various levels of organization, from the atomic-molecular to the cellular to the organ-organism level. The response of an organism to disturbances that are detrimental to structure and function generally begin at the level of organization where the primary injury has occurred. Detriment that occurs from simultaneous or sequential, or single or multiple interactions at a relatively low level of organization tends to be transferred to higher levels. However, at each level of organization there is a given probability of such detriment being removed according to the tolerance to injury that is peculiar to that level. There is thus a direct relationship between the frequency of injurious events at a lower level of organization, and the degree of structural complexity of the system at the high level at which such detriment is eventually manifested. The extent of structural disruption at any given level determines the degree of functional failure at that level. In the exposure of tissue to ionization radiation, the primary injury begins with energy deposition events (tracks or hits) consisting of many ionizations and excitations in localized clusters of submicroscopic dimensions at the atomic-molecular level of organization within the cell, and the cell is affected as a whole. The cell is the elementary unit of life and the sum of the individual cell responses determines the response of the tissue and the organism. Individual cell responses are nevertheless found to differ in type and degree depending on the absorbed dose. With decreasing values of absorbed dose to the tissue, the probability of a cell being hit by an energy deposition event decreases linearly. At very low values of absorbed dose to tissue, only a fraction of the total cell population experiences single hits and these are of different sizes. The size distribution or spectrum of these hits is invariant, independent of their total number over a considerable range at low dose levels and is determined only by the type and quality of the given radiation. The probability that a hit cell will suffer a given detriment such as a chromosomal aberration, gene mutation or death has been shown to increase in a sigmoid fashion with increasing hit size.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 7488974 TI - Planimetric volumetry of the prostate: how accurate is it? AB - Planimetric volumetry is used in clinical practice when accurate volume determination of the prostate is needed. The prostate volume is determined by discretization of the 3D prostate shape. The are of the prostate is calculated in consecutive ultrasonographic cross-sections. This area is multiplied by the distance between the cross-sections and the total volume is determined by summation of all contributions. Besides the quality of the automated outlining, the accuracy of this method depends on this intersection distance and on the angle of the scan plane with the probe. Also, the location of the first cross section is of influence. This paper describes the influences of these parameters on the accuracy of the volume determination using a simple prostate model. This theoretical influence is compared to clinical volume determinations using automated planimetric volumetry with different step sizes. From our data, it is concluded that a step size of 4 mm for planimetric prostate volume determination is a good compromise between investigation time and accuracy in a clinical setting. PMID- 7488972 TI - East German uranium miners (Wismut)--exposure conditions and health consequences. AB - Underground uranium mining was performed in East Germany after World War II on a large scale. East Germany was the main supplier of uranium for the Soviet Union. This review gives a historical summary and describes the broad spectrum of exposure to potential health hazards and the health consequences. Working conditions were very poor during the postwar years from approximately 1946-1955: there was drilling with air floating and a lack of forced ventilation. Dust levels were very high and there was a significant inhalative incorporation of alpha-radiating substances, mostly from short-lived radon progeny. However, long lived alpha-radiating substances such as uranium-238 contributed considerably to the radiation dose. There was also exposure to toxic chemicals, such as arsenic (in some mines) or crystalline silica, and a variety of other health hazards. From approximately 1956-1970, mining conditions improved: there was drilling with the addition of water and forced ventilation of the mines. As of approximately 1970, compliance with rules of industrial hygiene and international standards of radiation protection was evident. In 1990, uranium production was generally stopped. To date, more than 5,000 cases of bronchial carcinoma are accepted as compensable occupational diseases and more are expected. The extensive data from Wismut uranium mining could improve our understanding of a complex exposure situation resulting in a variety of health impairments other than lung cancer. PMID- 7488973 TI - HOXB gene expression and function in differentiating purified hematopoietic progenitors. AB - Intensive efforts have led to the development of methods for stringent purification of adult hematopoietic progenitor cells (HPCs), particularly from peripheral blood (PB). The purification procedure previously reported by our group (Science, 1990) provided a high HPC frequency, but yielded a low HPC recovery (< or = 5-10%). We therefore developed an improved purification methodology based on "potentiated" negative immunobead selection (Step IIIP) by addition of anti-CD45, -11a and -71 monoclonal antibodies (mAbs) to the previously utilized panel of mAbs. This simplified procedure consistently allows not only high level purification but also abundant recovery of early HPCs: the final Step IIIP cell population (0.95 x 10(6) cells/4 PB donors, mean value) features an 81% HPC frequency and a recovery of 45% of the initial HPCs. The purified HPCs bear the primitive HPC phenotype, i.e., they are consistently CD34+, largely CD33-/45RA-, and in part HLA-DR-/low/CD38-/low/Thy-1+. In optimized semi-solid culture, the purified erythroid/multipotent HPCs give rise to macroscopic colonies (10,000-150,000 cells/clone, > 0.5 mm size colonies). This purification methodology compares favorably with previously reported procedures in terms of combined HPC frequency and recovery: availability of a large number of highly purified, early HPCs will provide an experimental tool for analysis of the molecular/cellular basis of early hematopoiesis. We have investigated by reverse transcription-polymerase chain reaction (RT-PCR) the mRNA expression of homeobox B (HOXB) cluster genes in purified HPCs induced in liquid suspension culture to gradual erythroid or granulopoietic (largely eosinophilic) differentiation and maturation by differential growth factor (GF) stimulus. Only B3 is expressed in quiescent HPCs. After GF treatment B3 expression is enhanced in the initial 24 h and then through erythroid and granulopoietic differentiation and maturation. HOXB4 and B5 are induced at slightly later times and expressed through maturation in both lineages, while B6 is selectively induced in granulocytic differentiation. B2 is transiently expressed at low level in the granulopoietic pathway, while it is detected only in advanced stages of erythropoiesis; B7, B8 and B9 are essentially not detected. Functional studies were performed with antisense phosphorothioate oligomers to HOX mRNAs including: 1) anti-B3 oligomer (alpha-B3) treatment of purified HPCs induces a striking blockade of both erythroid and granulomonocytic colony formation, 2) alpha-B6 selectively and markedly inhibits granulomonocytic colony formation, 3) alpha-B4 and alpha-B5 cause a significant, less pronounced decrease of both colony types and finally, 4) alpha-B2 and alpha-B7, alpha-B9 exert little and no effect respectively. PMID- 7488975 TI - Measuring the mechanical properties of the lung in vivo with spatial resolution at the acinar level. AB - The alveolar capsule technique involves measuring subpleural alveolar pressure in the chamber of a plastic capsule glued to the perforated lung surface. However, measurements of alveolar pressure alone do not permit one to say whether the inhomogeneities that develop are due to changes in local lung resistance, elastance, or both. We have developed an extension of the alveolar capsule technique in which small broad-band oscillations in flow are applied through the capsule. At low oscillation frequencies the imposed flows travel all the way through the airway system, so the associated alveolar pressures then reflect the resistance of the pathway from a small local alveolar region out to the tracheal opening. At high frequencies flows propagate only a very short distance into the lung whereupon the dynamic relationship between flow and alveolar pressure gives a measure of local lung elastance. In this paper we show that considerations of canine lung anatomy suggest that the elastic and resistive quantities provided by the capsule oscillator technique pertain to a lung region of the order of a single acinus with its associated terminal bronchioles. This represents a greatly improved spatial resolution of lung mechanics over that which was possible before the development of the alveolar capsule oscillator technique. PMID- 7488976 TI - A new non-invasive approach for monitoring respiratory movements of sleeping subjects. AB - We have developed a minimally intrusive system to monitor respiratory movements of sleeping subjects. This system is based on a pressure transducer which measures the changes in air pressure inside an inflatable mattress on which the subject sleeps. Using a mechanical filter to protect the transducer against the large pressure changes due to sudden movements and subject weight, we can detect the more subtle movements of the subject's chest. This paper discusses the design of the monitoring system, including the design and modelling of the mechanical filter. PMID- 7488977 TI - Monitoring of renal pelvic pressure in patients with hydronephrosis. AB - A diagnostic method for the determination of obstructions in the pelvoureteral junction in patients with suspected idiopathic hydronephrosis is described and discussed. Two microtransducer catheters (MTCs) with infusion lumens are inserted under fluoroscopic control, one into the renal pelvis and the other in the perirenal space as a reference. The pressure values obtained are stored on a portable microcomputer system, carried by the ambulatory patients. Renal pelvic pressure, intra-abdominal pressure and differential pressure are calculated and displayed as time-pressure diagrams on a plotter. The pressures can be measured for long periods (3-24 h). Furthermore, steady state conditions can be evaluated during pelvic infusion of saline. The results from the pressure measurements are compared with clinical and radiological evaluations. Our results show that this method of long-term monitoring of intrapelvic pressure together with intra abdominal pressure contributes to the evaluation of patients with suspected pelvoureteral obstructions and complements other methods in the preoperative investigation. However, this study reveals difficulties with measurements of intra-abdominal pressure. Carrying the equipment was not inconvenient for the patients. PMID- 7488978 TI - Laser profiling: a technique for the study of prosthetic heart valve leaflet motion. AB - A detailed understanding of the stresses and strains developed in functioning flexible-leaflet valves is necessary if a durable, non-thrombogenic heart valve replacement is to be realized. A new experimental tool, laser profiling, is presented for the study of flexible-leaflet heart valve dynamics. Profiles of moving leaflet surfaces are obtained by projecting parallel sheets of laser light onto valve leaflets as the valves open and close in a mock circulatory loop. Two versions of laser profiling have been developed. In two-dimensional mode multiple profiles are generated on a fixed plane in space but at discrete intervals in time, whereas in three-dimensional mode multiple profiles are generated across the leaflet surface at (effectively) a single instant in time. Highlighted leaflet profiles are captured by camera and transferred to an image processing system for analysis. A simple algorithm permits digitized profiles to be reconstructed within a computer-aided design software package, providing detailed visualization and quantification of valve motion. Extensive validation studies have been performed using the Medtronic-Hall mechanical prosthetic heart valve. Laser profiling enables computer reconstruction of the rigid occluder to an accuracy of +/- 200 microns from a 0.7 ms exposure taken during the period at which the occluder moves with greatest velocity. The technique has been applied to investigate the leaflet dynamics of a bovine pericardial heart valve prosthesis. PMID- 7488979 TI - The feasibility of long-term fetal heart rate monitoring in the home environment using maternal abdominal electrodes. AB - It is well established that fetal and maternal electrocardiograms (ECGs) can be obtained from the maternal abdomen using standard surface electrodes, although this cannot be guaranteed. The unobtrusive and non-invasive nature of such monitoring lends itself naturally to the long-term ambulatory collection of this data on cardiac activity. By employing suitable algorithms it would then be possible to extract records of both fetal and maternal heart rate. This article presents results of the collection of raw electrophysiological signals, containing both fetal and maternal ECGs from a single volunteer from the 20th week of gestation until term. The significance of the data is that they were recorded by the mother herself in her own environment. Previously written software was then used to extract fetal and maternal heart rate data. These results demonstrate the feasibility of using this method for the long-term recording of fetal and maternal heart rate in the mother's normal surroundings. PMID- 7488980 TI - Who is responsible? PMID- 7488981 TI - Magnetic resonance spectroscopy of the human masseter muscle in nonbruxing and bruxing subjects. AB - The masseter muscles of six nonbruxing subjects (five men, one woman) and six bruxing subjects (four men, two women) were assessed during chewing by nuclear magnetic resonance spectroscopy (31P-NMR). The NMR spectra were collected on a GE Sigma 1.5T whole body magnet with a double-tuned 31P/1H surface coil. Two-minute trials of rest/chewing/rest were completed three times. Averaged spectra of inorganic phosphate, phosphocreatine, and three adenosine 5' triphosphate peaks were collected in each trial. Bruxing subjects had a lower concentration of total phosphate and phosphocreatine than nonbruxing (control) subjects at rest. Bruxing subjects increased their inorganic phosphate during chewing significantly less than control subjects. The pH levels during rest and during chewing were similar in both controls and bruxers. These preliminary results suggest that bruxing subjects exhibit an altered phosphate metabolism during rest and exhibit a different phosphate metabolism pattern during chewing as compared to nonbruxing subjects. PMID- 7488982 TI - Interleukin-6 in synovial fluid and HLA-DR expression in synovium from patients with temporomandibular disorders. AB - Interleukin-6-dependent mouse hybridoma cell line KD83 was used to test the biologic activity of interleukin-6 in synovial fluid from 37 patients with temporomandibular disorders. The results showed that the interleukin-6 level was greater than 100 U/mL in 13 of 18 patients with degenerative joint disease and in five of 12 patients with temporomandibular disc displacement. However, the interleukin-6 level was less than 100 U/mL (range, 20 to 75 U/mL) in all patients with masticatory muscle disorder. It has been found that degenerative joint disease tends to have acute and chronic stages, and interleukin-6 activity was probably related to the acute stage in the patients. Histologic studies of the synovium from seven patients with degenerative joint disease showed a variable degree of hyperplasia of the synovial lining cells and chronic inflammation in five of eight specimens. Immunostaining studies clearly showed the presence of significantly more HLA-DR-expressing cells (human leukocyte antigen-D-related) in synovium. Although it is unlikely that immune responses play an important primary role in initiating synovial inflammation and cartilage destruction, immune reactions may be one important factor in the maintenance and severity of some patients with temporomandibular disorders. PMID- 7488983 TI - Thermographic assessment of neuropathic facial pain. AB - Ongoing pain, intermittent sharp pain, or intermittent dull aching pain around the teeth can evoke the suspicion of tooth pathology. However, when no dental cause can be found clinically or radiographically, the differential diagnosis involving neuropathic pain and pulpal pathology is still a challenge. Neuropathic facial pains are still too often misdiagnosed as tooth pain of dental origin, resulting in unnecessary dental extraction or endodontic therapy. The purpose of this study was to determine if electronic thermography was able to differentiate neuropathic facial pains presenting as toothache from pulpal pathology. Electronic thermography was used to compare asymptomatic subjects and subjects with neuropathic facial pains. Asymptomatic subjects and subjects with trigeminal neuralgia, pre-trigeminal neuralgia, and pulpal pain without periapical pathology showed no thermographic difference in the territory of the pain complaint when compared to the opposite nonpainful side. Patients with sympathetically maintained traumatic trigeminal neuralgia (atypical odontalgia) and half of the group with sympathetically independent traumatic trigeminal neuralgia presented with "hot" thermograms. The other half of the patients with sympathetically independent traumatic trigeminal neuralgia displayed "cold" thermograms in the area of their pain complaints. Electronic thermography was the least selective test for the group showing "cold" thermogram patterns (80% agreement with the thermographic characterization criteria). These data suggest that electronic thermography may be helpful in differentiating neuropathic pains from pulpal pathology. PMID- 7488984 TI - History of treatment received by patients with TMD: a preliminary investigation. AB - A retrospective investigation of 257 patient records from a university-based facial pain center for patients with temporomandibular disorders examined the "natural history" of patients with temporomandibular disorders for consultation and treatment of their complaints. Data were obtained on a variety of demographic variables, referral patterns, types of providers seen for consultation, and the types of diagnostic tests ordered by these providers. Results showed that patients with temporomandibular disorders see more than three providers prior to their referral to a tertiary care center, that a considerable proportion of those providers are physicians, that patients reported undergoing a variety of diagnostic tests, and that patients reported receiving a variety of diagnoses for their condition. Results also showed that patients who initially consulted a physician were somewhat more likely to be referred to another physician, while patients who consulted a dentist were highly likely to be referred to another dentist. PMID- 7488985 TI - Stress distributions in the TMJ during clenching in patients with vertical discrepancies of the craniofacial complex. AB - This study was designed to investigate stresses in the TMJ during clenching in patients with skeletal discrepancies in the vertical direction. A three dimensional model of the mandible including the TMJ was used for finite element analysis for the stresses. The model, referred to as a standard model, consists of 2,088 nodes and 1,105 solid elements, comprising the cortical and cancellous bones, articular disc and cartilage layer, and periodontal ligament. The standard model was modified by varying the gonial and mandibular plane angles to simulate vertical discrepancies between the maxilla and mandible observed in open and deep bites. Stresses were analyzed on the surfaces of the condyle, the glenoid fossa, and the articular disc, and the values were compared to those found with the standard model. Stresses increased substantially for the condyle, the glenoid fossa, and the articular disc with greater gonial and mandibular plane angles, and those changes were more obvious in association with the divergent mandibular plane. Thus, the nature of stress distributions in the TMJ was substantially affected by vertical discrepancies of the craniofacial skeleton. It is also suggested that these changes in stresses produce a lack of biomechanical equilibrium in the TMJ, which may have some association with temporomandibular disorders. PMID- 7488986 TI - The relationship between forward head posture and temporomandibular disorders. AB - This study investigated the relationship between forward head posture and temporomandibular disorder symptoms. Thirty-three temporomandibular disorder patients with predominant complaints of masticatory muscle pain were compared with an age- and gender-matched control group. Head position was measured from photographs taken with a plumb line drawn from the ceiling to the lateral malleolus of the ankle and with a horizontal plane that was perpendicular to the plumb line and that passed through the spinous process of the seventh cervical vertebra. The distances from the plumb line to the ear, to the seventh vertebra, and to the shoulder were measured. Two angles were also measured: (1) ear-seventh cervical vertebra-horizontal plane and (2) eye-ear-seventh cervical vertebra. The only measurement that revealed a statistically significant difference was angle ear-seventh cervical vertebra-horizontal plane. This angle was smaller in the patients with temporomandibular disorders than in the control subjects. In other words, when evaluating the ear position with respect to the seventh cervical vertebra, the head was positioned more forward in the group with temporomandibular disorders than in the control group (P < .05). PMID- 7488987 TI - Degenerative disease of the temporomandibular joint. AB - Progression of degenerative joint disease is dependent on the underlying pathologic and/or reactive processes involved that, in general, compromise tissue adaptability. A review of clinical and experimental literature relating to degenerative joint disease is presented. Epidemiology, pathogenesis, diagnosis, treatment, and prognosis are described with particular emphasis given to the temporomandibular joint. This article describes factors affecting the temporomandibular joint remodeling/degeneration parity and presents rationale for approaches to diagnosis and treatment. PMID- 7488988 TI - Reliability of clinical findings in temporomandibular disorders. AB - The aim of the present investigation was to study the interexaminer reliability of orthopedic tests and palpation techniques routinely used in the clinical diagnosis of disorders of the masticatory system. The tests were performed by a dentist and a physiotherapist, who both used the tests routinely when examining patients with temporomandibular disorders. Seventy-nine patients participated in this study. In the analysis, percentage agreement, intraclass correlation, and Cohen's kappa were used. The interexaminer reliability of the tests measuring maximal active mouth opening and registration of clicking during active mouth opening was high. The interexaminer reliability was fair for the tests measuring the intensity of pain during active movements and moderate for tests recording joint sounds (kappa = 0.47 to 0.59). There was high interobserver agreement on several items of the traction and translation tests, although the kappa values were low. The interexaminer reliability of the multitest scores for compression was substantial for joint sounds (kappa = 0.66) and fair for pain (kappa = 0.40). The interexaminer reliability of the multitest scores for muscle palpation and joint palpation was moderate (kappa = 0.51) and fair (kappa = 0.33), respectively. It can be concluded that most variables determined during active movements can be measured with satisfactory reliability, whereas variables for other tests are not measured with the same reliability on the basis of the kappa scores. The main symptoms of temporomandibular disorders can be evaluated reliably with multitest scores. It is recommended that clinicians calibrate their techniques regularly to improve the reliability of results in daily practice. PMID- 7488989 TI - A randomized clinical trial of intraoral soft splints and palliative treatment for masticatory muscle pain. AB - Thirty subjects seeking treatment for masticatory muscle pain at a university based TMJ clinic were randomly assigned to soft-splint, palliative-treatment, and no-treatment groups. After 4 to 11 weeks of treatment, subjects were evaluated for changes from their baseline levels of symptoms, maximum pain-free opening, pain thresholds measured by a pressure algometer, and occlusal contacts. With the use of the multivariate analysis of variance and analysis of covariance, the results suggest that the soft-splint group had statistically significant improvement (P < .01), the palliative-treatment group had improvement that was not statistically significant, and the no-treatment group had a slight aggravation of symptoms. The soft-splint group had fewer occlusal contact changes assessed with shimstock compared to the palliative-treatment and no-treatment groups. The findings of this study suggest that the soft splint is an effective short-term treatment for reducing the signs and symptoms of masticatory muscle pain in patients, and the soft splint does not cause occlusal changes. PMID- 7488990 TI - Treatment of open fractures of the foot and ankle: a preliminary report. AB - A retrospective study was performed on 33 patients who sustained isolated open fracture injuries to the foot and ankle. A wide variety of open-fracture injuries of the foot and ankle are documented. Of the 33 cases, only 2 became clinically infected, translating into an infection rate of 6.06%. A literature review of open-fracture injuries is presented along with the preliminary results of the retrospective study. PMID- 7488991 TI - Surgical considerations for hematogenous osteomyelitis. AB - Surgical treatment of hematogenous osteomyelitis is uncommon in the literature. Indications for surgical decompression, along with a review of the literature, is presented. A case history of a 7-year-old male with hematogenous osteomyelitis of the first metatarsal treated by surgical decompression and 12-month follow-up is discussed. PMID- 7488992 TI - Lower extremity manifestations of Vibrio vulnificus infection. AB - Vibrio vulnificus is a potentially lethal marine bacterium that has not been previously described in podiatric literature. A review of the microorganism's characteristics, susceptible patient population, and lower extremity manifestations of infection is presented. V. vulnificus is found as part of the normal flora of the Gulf of Mexico, Atlantic, and Pacific coastal waters and is often isolated from the filter feeding shellfish of these regions. Its pathogenicity is generally reserved for the immunocompromised host, and is specifically related to disease states which exhibit high serum iron levels. V. vulnificus infections present in two distinct clinical syndromes: primary sepsis secondary to raw oyster ingestion, or localized infection from wound exposure to V. vulnificus-inhabited salt water. Both syndromes demonstrate characteristic skin lesions of the trunk and extremities that present as hemorrhagic bullae and progress to necrotic ulcerations. Although V. vulnificus infection is rare, its extreme virulence in patients suffering from a chronic disease process and its manifestation of characteristic lower-extremity lesions require the podiatric physician to be able to recognize and treat such a condition. PMID- 7488993 TI - Avascular necrosis of the hallucial sesamoids. AB - The authors present a literature review and systematic approach to the diagnosis and treatment of avascular necrosis of the sesamoids of the flexor hallucis brevis tendon. Renander, in 1924, was one of the earliest authors to call attention to this condition. Since that time, many other authors have written about this entity, some even questioning its existence. Many different treatment regimes have been postulated, encompassing both the conservative and surgical modalities. Most literature advocates attempted conservative treatment followed by surgical excision, only if conservative methods fail. PMID- 7488994 TI - Orthosorb: a case of foreign-body reaction. AB - The authors present a previously unreported foreign-body reaction to poly-p dioxanone pin fixation of a distal first metatarsal head osteotomy. The reaction was diagnosed by fine-needle aspiration under fluoroscopic guidance and computed tomography scan. The patient responded to conservative therapy and time, but, still has residual pain 11 months after the surgery. PMID- 7488996 TI - Talar fracture/dislocation in the adolescent patient. AB - The authors report an 18-year-old patient who sustained a two-part injury involving his right ankle. The result of this traumatic episode was a fracture/dislocation of the talus. An interesting aspect of this injury is that the body of the talus dislocated laterally, distal to the fibular malleolus. Closed reduction was attempted and subsequent open reduction with internal fixation was performed. The patient was followed for 9 months, during which time a series of radiographs was taken. Nine months postoperatively, Hawkins sign was negative indicative of avascular necrosis. In review of the literature, no fractures with lateral dislocations of this nature were found. PMID- 7488995 TI - Cuneiform and cuboid wedge osteotomies for correction of residual metatarsus adductus: a surgical review. AB - Metatarsus adductus is a deformity in which the metatarsals are deviated in the transverse plane when compared with the longitudinal axis of the lesser tarsus. A brief review of the deformity is presented including the causes, radiographic analysis, conservative treatment, and a surgical approach. A series of cases of metatarsus adductus is presented in which a proximal surgical approach consisted of cuneiform and cuboid wedge osteotomies with staple fixation. The described procedure has advantages over the traditional metatarsal wedge osteotomies by providing large cancellous bone-to-bone interfaces for excellent healing potential, easier fixation, sagittal plane stability, and avoidance of the physis at the base of the first metatarsal. PMID- 7488997 TI - A variable first metatarsal distal "L" osteotomy with adductor tendon transfer. AB - The authors present a modified approach to addressing hallux abducto valgus. We have found that a combination of an adductor tendon transfer through a distal metaphyseal osteotomy is useful in the elderly patient with a high intermetatarsal angle, who might otherwise require a proximal-type procedure for optimal correction. The results of seven patients who underwent this surgical procedure are presented as a preliminary report. PMID- 7488998 TI - Transfibular osteotomy in the correction of ankle joint incongruity. AB - The mechanics involved in Danis-Weber-types B and C ankle fractures may allow the fibula to seek a shortened and externally rotated position following injury. Residual talar instability secondary to fibular malalignment may ensue if proper anatomic restoration is not achieved at the time of reduction. Evaluation of ankle incongruity may take the form of clinical, radiographic, and diagnostic techniques. Three case studies are presented in which fibular osteotomies were utilized to restore proper anatomic alignment and function to the ankle mortise. PMID- 7488999 TI - Endoscopic plantar fasciotomy: a multi-surgeon prospective analysis of 652 cases. AB - The authors have analysed 652 cases in which an endoscopic plantar fasciotomy was performed. These procedures were performed and clinically evaluated by 25 different surgeons, who completed an accredited cadaveric training symposium. In connection with each endoscopic plantar fasciotomy performed, the participating surgeons provided patient data on evaluation forms which were subsequently submitted to the authors. The results of this study demonstrate that the endoscopic plantar fasciotomy has become an established procedure with a high rate of success. Additionally, the procedure is minimally traumatic, and allows patients to resume regular activities quickly with less pain or discomfort than traditional methods of surgical management. PMID- 7489000 TI - Imaging of pedal soft tissue neoplasms. AB - The authors discuss various radiographic imaging techniques for pedal soft tissue lesions. These include xeroradiography, angiography, ultrasound, computed tomography, and magnetic resonance imaging. Illustrative cases of each modality, as well as an evaluation of advantages and disadvantages, are presented. PMID- 7489001 TI - [The mechanisms of the recovery of nerve ending functions during the reinnervation of skeletal muscle]. AB - At the experiments on the frog cutaneous-pectoris muscle the nerve terminal functions in course of reinnervation process were investigated by electrophysiological and morphological methods. At the 20-25th days after the nerve crushing the nerve terminal response, which reflect the nerve terminal currents, formed the propagated action potential, were restored and the arising of evoked transmitter secretion occurred. The regenerating terminals are characterized by a low amplitude and altered shape of responses, by small velocity of the excitation propagation and the low level of evoked transmitter release. The 4-aminopyridine effect at the new formed nerve terminals was quite another, than at the intact nerve terminals. This data concluded, that the membrane of regenerating nerve terminal have a low density and a small gradient of sodium channels along the terminal and have not a calcium-activated potassium channels. It was proposed, that different kinds of channels are inserted into the nerve endings membrane at the different periods of the reinnervation process. PMID- 7489002 TI - [The effect of transcranial electrostimulation on initial alcoholic motivation in rats]. AB - The effect of transcranial electrical stimulation (TES) was found to inhibit alcohol intake in rats used to ethanol, whereas in those unused to ethanol the alcohol intake increased after the TES. The TES effect seems to be connected with the difference in opioid system conditions in animals with different spontaneous alcohol motivation. PMID- 7489003 TI - [The effect of the age of female rats on the brains of their progeny]. PMID- 7489004 TI - [Circadian fluctuations in the temporal organization of forced swimming in rats]. PMID- 7489005 TI - [The effect of lead on the Ca2+, Mg2+-ATPase and Na+, K+-ATPase activities of the plasma membrane of smooth-muscle cells]. PMID- 7489006 TI - [The problems of human ecological physiology]. PMID- 7489007 TI - [A new method for correcting alcoholic motivation]. PMID- 7489008 TI - [A device for the artifact-free lead take-off of brain electrical activity in exposure to a UHF field in free-ranging rats]. PMID- 7489009 TI - [The role of the individual subtypes of the brain adreno- and dopaminergic receptors in regulating the function of the hypothalamo-hypophyseal-testicular system]. AB - Activation of different subtypes of adrenergic (AR) and dopamine receptors (DR) and its effect on the blood testosterone level, were studied. Intraventricular administration of selective agonists mesatone and clonedine as well as dobutamine increased the testosterone level in the blood, where as clenbuterol exerted no such effect. Activation of D1-DR by SKF 38393 in ZI and PVGM elevated the testosterone level as well. Activation of D2-DR by LY 171555 in ZI exerted no effect and in the PVGM even inhibited the testosterone level. PMID- 7489011 TI - [The current status of the study of the physiology of the autonomic nervous system in Russia]. PMID- 7489010 TI - [The long-term depression of the function of the hypothalamo-hypophyseal adrenocortical system in rats]. AB - Hypothalamo-pituitary-adrenocortical axis reactivity was studied in vivo and in vitro in 7 days after single administration of 30 mg/100 g body weight of hydrocortisone in rats. There were a blockade of stress-induced decrease of CRF 41 content in median eminence, and failure of increase of ACTH and corticosterone levels in the blood to stress in 7 days after hydrocortisone administration. A decrease of reactivity to ACTH was shown during incubation of adrenals from rats treated with hydrocortisone. 300 and 400 mg/100 g body weight of corticosterone s. c. imitates a stress-induced increase of corticosterone in the blood and can be used as replacement therapy. We conclude that high supraphysiological doses of glucocorticoids results in a decrease of reaction of hypothalamus, pituitary and adrenals to stress. PMID- 7489012 TI - [The neurophysiological correlates of improvement in the cognitive characteristics of monkeys with modification of the NMDA-ergic structures of the prefrontal cortex]. AB - The effect of the NMDA glutamate agonist on cognitive characteristics and short term memory (SM) was studied in rhesus monkeys. The NMDA effect manifested itself in doubling the SM term and in changing of the unit activity in the prefrontal cortex. The NMDA also induced a considerable increase in the cross-correlation coefficients between the unit responses in visual and prefrontal cortex. The data obtained suggest that the glutamatergic structures of prefrontal cortex take part in the processes of visual recognition and the SM in monkeys. The effect seems to improve the short-term memorizing of a visual information. PMID- 7489013 TI - [Adenylate cyclase and 5'-nucleotidase activities in the sensorimotor and limbic structures of the rat brain after training in a manipulative skill]. AB - The activities of adenylate cyclase (AC) and 5'-nucleotidase (NT) were studied in the limbic (amygdala, hippocampus) and sensorimotor (cortex, striatum) brain structures of three rat groups: control (without training), not well and well learning to perform movements with pushing the operandum. It was found, that after training the activity of AC was decreased in all structures studied. Moreover, in the cortex and the striatum the decrease was more pronounced in the group of well learning rats, while in the amygdala--in not well learning ones. The activity of NT in all brain structures, excepting the striatum, was more significantly decreased in the rats with low ability to learn the movements with prolonged pushing. Only in the striatum the increase of NT activity (initially the lowest in the control animals--1.0 +/- 0.04 microgram P(i)/mg protein/min) up to 1.3 +/- 0.1 in not well and up to 2.0 +/- 0.1 in well learning animals was found. The interhemispheric [correction of intrahemispheric] asymmetry of AC activity in the cortex and of NT in the hippocampus was revealed. Thus, the changes of the activity of the enzymes, participating in the biogenesis of adenosine (NT) and c-AMP (AC) are different in the limbic and sensorimotor structures dependently on learning ability of rats. The increase of NT activity after learning only in the striatum could reflect a specific role of purinergic system of this brain structure in the regulation of sensory controlled movements. PMID- 7489014 TI - [The reactions of rat respiratory neurons to a hypoxic stimulus]. AB - Hypoxia was shown to lead to hyperpnea and an increase in the breathing rate in rats due to shortened expiration. The unit activity of respiratory neurons was enhanced. The activating effects from the carotid chemoreceptors on the respiratory responses seem not to be uniform. PMID- 7489015 TI - [The neuropharmacological correction of cognitive processes disordered as a result of the extirpation of fields AI-AII and Ep in cats]. PMID- 7489016 TI - [The redistribution of calcium ions and organic substances in pregnant rats under extreme conditions]. AB - It was found that under water deprivation the liver, spleen, white and brown fat decreased in weight in rats whereas the weight of the heart, kidneys and uterus with fetuses did not differ from the control. In pregnant rats, water deprivation led to involution of thymus and hypertrophy of adrenals, decrease in the calcium ions content in the liver and kidney and reducing of the liver and some other organs weight. A similar picture was observed during space flight in pregnant rats. The data obtained suggest a redistribution of organic and inorganic substances under water deprivation. PMID- 7489018 TI - [The effect of continuous unmodulated microwaves on the electrical activity of the cerebral cortex, the heart beat frequency, respiration and general condition of the rabbit]. PMID- 7489017 TI - [The effect of sodium nitrate on the digestive enzyme activity of the small intestine, liver and kidneys of rats]. AB - The activity of digestive enzymes was reduced both in the small intestine mucosal layer and in the postepithelial ones. The reduction was due to sodium nitrate administration and was more obvious in the rats fed with proteins. The response of the enzyme systems of the liver and kidney was heterogeneous. PMID- 7489019 TI - [The adrenergic mechanisms of intravascular pressure stabilization when the volume velocity of the blood flow is altered]. AB - Experiments in cats revealed that the vasoconstrictor effect of norepinephrine and propranolol involved an increase in extensibility of vessels. Preliminary beta-adrenoceptor blockade decreased both the vessels' extensibility and their ability to resist against growing pressure in acceleration of the blood flow volume velocity. The role of adrenergic systems in intravascular pressure stabilisation in acceleration of the blood flow volume velocity is emphasized. pressure stabilisation in acceleration of the blood flow volume velocity is emphasized. PMID- 7489020 TI - [An analysis of the afferent flows from the glabrous skin receptors of cat foot pads during thermomechanical effects]. AB - Cooling of the cat paw skin augmented the afferent activity in the n.n. plantaris whereas warning of the paws exerted an opposite affect. The code of thermomechanical sensory signals depended on the physical-mechanical condition of the skin. PMID- 7489021 TI - A neuroendocrine reappraisal of the dual-centre hypothesis: its implications for obesity and insulin resistance. PMID- 7489022 TI - Improved prediction formula for total body water assessment in obese women. AB - OBJECTIVE: To validate an improved formula for the calculation of total body water (TBW) from impedance values in obese women. SUBJECTS: A sample of 37 overweight females BMI ranging from 29.5 to 44.1 kg/m2. MEASUREMENTS: The reference measurements for TBW were obtained by deuterium oxide dilution. Body height (h), hip circumference (C) as well as body impedance at 100 kHz (Z100) were also measured. RESULTS: The values obtained for h, C and Z100 were introduced in the calculations and the following developed formula for total body water was determined: TBW = 0.069 [hC2/(4 pi .Z100)]+19.671. The validity of this formula was assessed through the statistical test of Bland and Altman which we also performed for other formulas containing the weight and/or the height. CONCLUSION: We found that the accuracy of the prediction of TBW by our formula, although lower than those formulas using body weight, is higher than that which takes into account only body height. We conclude that body volume, expressed through body height and hip circumference, is a good choice of parameter in bioelectrical impedance measurements. The new approach can be useful in clinical settings for repeated monitoring of obese women, during diet-restricted treatment. PMID- 7489024 TI - Reproducibility of 24-h energy expenditure, substrate utilization and spontaneous physical activity in obesity measured in a respiration chamber. AB - OBJECTIVE: Determination of within-subject day-to-day coefficients of variations (CV) of 24-h measurements of spontaneous physical activity (SPA), macronutrient oxidation, energy expenditure (EE) and heart rate (HR). DESIGN: Two 24-h measurements in respiration chamber, separated by 14 days of strict weight stability. SUBJECTS: 22 healthy obese subjects, 15 females and seven males (age 27-64 y, BMI: 30-39 kg/m2). MEASUREMENTS: SPA assessed by microwave radar, substrate oxidation and EE measured by indirect calorimetry and heart rate measured by telemetry. RESULTS: For 24-h measurements CV was 0.8% for RQ, 6.7% for fat oxidation, 5.6% for carbohydrate oxidation, 14.5% for protein oxidation, and 4.3% for heart rate. SPA assessed by microwave radar was also reproducible (CV = 9.7%). Within-subject day-to-day coefficients of variations (CV) were 1.5 and 1.8% for 24-h EE and sleeping EE, and 4.1% for RMR. Both on day 1 and 14, 93.4% of the variation in 24-h EE could be explained by differences in fat-free mass (92.3 and 92.4%, respectively) and SPA (1.1 and 1.0%), respectively). Also the algorithm and correlation between 24-h RQ and 24-h energy balance were highly reproducible (r = 0.56, P < 0.01 and r = 0.48, P < 0.05). CONCLUSION: Both macronutrient oxidation, SPA and EE in obese patients can be determined with high reproducibility in a respiration chamber. PMID- 7489023 TI - Hepatic mitochondrial respiration and transport of reducing equivalents in rats fed an energy dense diet. AB - OBJECTIVE: To measure hepatic mitochondrial respiration as well as the ability of hepatic mitochondria to transport reducing equivalents by shuttle systems in rats fed an energy dense diet. DESIGN: Rats were fed a control (CD) or energy dense (ED) diet for 15 days. MEASUREMENTS: State 3 and State 4 oxygen consumption were measured in isolated mitochondria using glutamate+malate or pyruvate+malate as substrate. We also measured malate-aspartate shuttle activity and mitochondrial alpha-glycerophosphate dehydrogenase activity. RESULTS: ED rats, in comparison with CD rats, showed a significantly greater energy intake without a corresponding greater body weight gain and carcass lipid content. Compared to CD rats, ED rats also showed a significant increase in resting metabolic rate, which was abolished by propranolol. Hepatic mitochondrial state 3 respiration using glutamate+malate or pyruvate+malate as substrate as well as malate-aspartate shuttle activity significantly decreased, while mitochondrial alpha glycerophosphate dehydrogenase significantly increased in ED rats compared to CD rats. CONCLUSIONS: Mitochondrial NADH oxidation is reduced and a greater fraction of cytoplasmic NADH can be transported to the mitochondria through the alpha glycerophosphate shuttle and oxidized through the respiratory chain from complex II onwards. This mechanism could lead to a decrease in hepatic metabolic efficiency which is in line with the increased energy expenditure occurring in rats fed an energy dense diet. PMID- 7489025 TI - Interrelationship of left ventricular mass, systolic function and diastolic filling in normotensive morbidly obese patients. AB - OBJECTIVE: To determine the interrelationship of left ventricular (LV) mass, systolic function and diastolic relaxation in morbidly obese subjects. METHOD: We obtained echocardiograms (M-mode, two dimension) and cardiac Doppler studies (pulse wave, continuous wave colour flow) on 50 subjects whose actual body weight was > or = twice ideal body weight. LV mass/height index was calculated from echocardiographic data (Penn Convention). LV systolic function was assessed by calculating LV fractional shortening. LV diastolic filling was assessed by measuring the transmitral Doppler E/A ratio and the transmitral E wave deceleration time. RESULTS: There were significant positive correlations between LV mass/height index and the LV internal dimensions in diastole, systolic blood pressure, LV end-systolic wall wall stress and the transmitral E wave deceleration time. There were significant negative correlations between LV mass/height index and both LV fractional shortening and the transmitral Doppler E/A ratio. There were significant negative correlations between LV fractional shortening and the LV internal dimension in diastole, systolic blood pressure LV end-systolic wall stress and the transmitral E wave deceleration time. There was a significant positive correlation between LV fractional shortening and the transmitral Doppler E/A ratio. There were significant positive correlations between the transmitral E wave deceleration time and LV internal dimension in diastole, systolic blood pressure and LV end-systolic wall stress. There were significant negative correlations between the transmitral Doppler E/A ratio and the aforementioned variables. CONCLUSIONS: Unfavourable alterations in LV loading conditions contribute to the development of LV hypertrophy and impairment systolic dysfunction in morbidly obese subjects. Increasing LV mass and altered loading conditions may synergistically contribute to impairment of LV diastolic filling in such individuals. PMID- 7489026 TI - Glucose utilization in morbidly obese subjects before and after weight loss by gastric bypass operation. AB - OBJECTIVE: To investigate the effect of weight reduction induced by gastric bypass operation (GBO), on the peripheral resistance to insulin in extremely obese subjects. DESIGN: A three-stage euglycemic clamp was applied to assess the metabolic clearance rate of glucose (g-MCR) in lean controls and in obese subjects, prior to and 6-12 months post operation. SUBJECTS: Six obese subjects (four obese normoglycemics-ON and two obese non-insulin dependent diabetics-OD) before and after GBO and six healthy, lean controls (LC) were compared. MAIN OUTCOME MEASURES: Body mass index after GBO, metabolic clearance rate of glucose under increasing insulin concentrations. RESULTS: GBO resulted in a significant change in body mass index from a pre-operation value (mean +/- s.d.) of 45.0 +/- 8.5 to 30.4 +/- 5.9 kg m-2 and remained significantly greater than controls (23.3 +/- 2.3 kg m-2). Glucose MCR increased from a mean baseline value of 3.0 +/- 1.6 to 6.7 +/- 3.9 ml kg-1 min-1 at post GBO (P) (P < 0.02). Similar effects were obtained under the two higher insulin concentration. CONCLUSIONS: GBO resulted in a massive reduction in BMI, that still remained above normal controls. This could explain the effect of weight reduction to decrease, but not completely reverse, peripheral resistance to insulin associated with obesity. PMID- 7489028 TI - The validity of recalled weight among younger women. AB - METHOD: The authors compared recalled weight at age 18 and self-reported current height among 118 participants in the Nurses' Health Study II cohort, aged 25-42 years in 1989, with records from physical examinations conducted at college or nursing school entrance. RESULTS: The correlation between recalled and measured past weight was 0.87 and between reported current height and measured past height was 0.94. Mean body mass index (BMI) values were 21.6 kg/m2 for BMI calculated using recalled weight and 22.1 kg/m2 using weight from medical records; the correlation was 0.84. Participants slightly under-reported weight at age 18 (mean difference = 1.4 kg). CONCLUSION: The validity of recalled weight at 18 years of age and self-reported height as an estimate of height at age 18 appears to be high among women in this cohort. PMID- 7489027 TI - Social variation in attitudes to obesity in children. AB - OBJECTIVE: To evaluate variations in children's attitudes to obesity in relation to age, gender and social class. DESIGN: Evaluation of sex, age and social class differences in judgements of figure drawings varying in body shape. SUBJECTS: 180 children aged from 4-11 from six schools, varying in the social background of their pupils. MEASUREMENTS: Self ratings of body shape, choices of ideal body shape now and in adulthood from a range of line drawings of figures varying in fatness, and behavioural and personality stereotype judgements of illustrations of thin, average and fat children. RESULTS: Few children in this age group were fat and few felt too fat; though a substantial minority felt too thin. Girls selected thinner ideal figures than boys. Children demonstrated strongly negative attitudes to obesity, which were less favourable among those who were older and from the higher social status schools. CONCLUSION: There are significant social variations in attitudes to obesity which might be important in understanding variations in the prevalence of obesity and weight control practices. PMID- 7489029 TI - Influence of macronutrients on adiposity development: a follow up study of nutrition and growth from 10 months to 8 years of age. AB - OBJECTIVE: To investigate the relationship between early nutrient intake and adiposity development. DESIGN: A follow up study of nutrition and growth carried out in a sample of 112 French children from 10 months to 8 years of age. MEASUREMENTS: Nutritional intakes at the age of 2 years and anthropometric measurements: Body Mass Index (BMI), subscapular and triceps skinfolds at the age of 8 years, and age at adiposity rebound assessed on the basis of BMI development. RESULTS: The BMI at the age of 8 years is positively correlated with energy intake at the age of 2 years, but this correlation becomes non significant after adjustment for BMI at 2 years. Protein (% of energy) intake at the age of 2 years is positively correlated with BMI and subscapular skinfold at 8 years after adjustment for energy intake at 2 years and parental BMI. The percentage of protein at 2 years is negatively associated with age at adiposity rebound, i.e. the higher the protein intake at 2 years, the earlier the adiposity rebound and the higher the subsequent BMI level. CONCLUSION: Protein at the age of 2 years is the only nutrient intake associated with fatness development pattern. A high protein intake increases body fatness at 8 years of age, via an early adiposity rebound. The association between protein intake and obesity is consistent with the increased stature and accelerated growth of obese children. A high fat low protein diet (such as human milk) is adapted to high energy demand for growth in early childhood. Our results suggest that high protein diet early in life could increase the risk of obesity and other pathologies later in life. PMID- 7489031 TI - Abdominal fat distribution and coronary heart disease risk factors in men waist/height ratio as a simple and useful predictor. AB - OBJECTIVE: To determine whether waist/height ratio is a useful predictor for coronary heart disease (CHD) risk factors in men. DESIGN: An epidemiologic study comparing relationships between waist/height ratio, body mass index, waist/hip ratio and CHD risk factor levels [continuous variables--systolic and dialostic blood pressure, fasting blood glucose, hemoglobin A1c (HbA1c), triglyceride, cholesterol, HDL cholesterol values] and the risk factor morbidity index (sum of the risk factor scores for hypertension, abnormal glucose tolerance, hypertriglyceridemia, hypercholesterolemia and low HDL cholesterol--one point each if present). SETTING: A health examination facility within a general hospital. SUBJECTS: 3131 men underwent routine health examination. RESULTS: Body mass index, waist/hip ratio and waist/height ratio were significantly associated with all of the risk factor levels and with the risk factor morbidity index according to the result of simple regression analysis. Multiple regression analysis for waist/hip ratio and body mass index showed that both of them were also significantly associated with all of the risk factor levels and with the risk factor morbidity index, except HbA1c levels with body mass index. According to the results of multiple regression analysis for waist/height ratio and body mass index, body mass index was not significantly associated with fasting blood glucose, HbA1c, cholesterol levels or the risk factor morbidity index whereas waist/height ratio was significantly associated with all of the variables. On the other hand, while waist/height ratio was significantly associated with all of the variables, waist/hip ratio was not significantly associated with most of the variables when waist/height ratio and waist/hip ratio were compared. CONCLUSION: Waist/height ratio, an index of abdominal obesity, may be a better predictor of multiple CHD risk factors in men than waist/hip ratio in mass epidemiologic studies. PMID- 7489030 TI - Self-reported versus measured height, weight and body mass index amongst 16-17 year old British teenagers. AB - OBJECTIVE: To examine the relationships between reported and measured height and weight in a teenage population group, and to assess the impact this may have on estimates of overweight. DESIGN: Data were taken from a sample of teenagers from the 1970 Birth Cohort Study. Multivariate normal regression was used to model differences between self-reported and measured height and weight, using both BMI and a number of other personal and demographic variables to examine influences on reporting differences. RESULTS: Tall, thin individuals were more likely to under report their height and shorter, fatter individuals to overestimate their height and under-estimate their weight. Self-reported height and weight data when used to calculate BMI would result in a lower estimate of overweight teenagers. Self assessment of body fatness, (but no other personal or demographic variable), was influential on the height and weight reporting of females in this study. CONCLUSION: Self-reported height and weight data from a teenage population should be used with caution, particularly if classifying individuals by BMI or when using weight measurements to estimate energy requirements. PMID- 7489032 TI - The hormone-sensitive lipase (LIPE) gene located on chromosome 19q13.1-->13.2 is not duplicated on 19p13.3. AB - The existence of a DNA polymorphism at the hormone-sensitive lipase locus could be of great interest for genetic analysis of obesity and related disorders since hormone-sensitive lipase is the rate-limiting enzyme of adipose tissue lipolysis and therefore plays a key role in energy metabolism. The polymorphic dinucleotide repeat D19S120 was identified within a human genomic clone selected with a rat hormone-sensitive lipase cDNA. This marker was subsequently localized to the short arm of chromosome 19 (p13.3) whereas human hormone-sensitive lipase (LIPE) had been mapped to the long arm of chromosome 19 (q13.1-->13.2). A duplication of the hormone-sensitive lipase gene or the presence of a pseudogene could explain the discrepancy. Cosmids from the two regions were analyzed in Southern blot experiments. A human adipose tissue hormone-sensitive lipase full-length cDNA probe hybridized only to cosmids from the 19q13.1-->13.2 region whereas the D19S120 amplicon probe hybridized only to cosmids from the p13.3 region. These data show that the occurrence of gene duplication or the presence of a pseudogene on the short arm of chromosome 19 is very unlikely and that D19S120 is unrelated to the hormone-sensitive lipase gene. PMID- 7489033 TI - Gallstone formation in obese women treated by a low-calorie diet. AB - This study assessed the incidence of gallstone formation in 47 obese women who consumed a low-calorie diet (LCD) for the first 16 weeks of a 26-week weight loss program. The LCD consisted of four daily servings of a liquid diet combined with an evening meal of a pre-packaged dinner entree and provided approximately 925 kcal/d. Six of the 47 patients (12.8%) displayed gallstones at week 17, as determined by sonography. Five patients were asymptomatic when followed for up to 48 weeks. The sixth, however, reported severe abdominal pain 30 weeks after beginning treatment and required a cholecystectomy. Patients who developed gallstones, as compared with those who did not, had significantly higher baseline triglyceride and total cholesterol levels and had a significantly greater rate of weight loss. Results of this study indicate that an increased risk of gallstones is not limited to very-low-calorie diets and that the incidence of this complication should be assessed in persons who consume popular over-the-counter meal replacement plans. PMID- 7489035 TI - Pain in the ICU: fact or fiction? PMID- 7489034 TI - Associations of old and new indices of adiposity with a group of hemodynamic and metabolic risk factors. PMID- 7489036 TI - Postoperative atelectasis and pneumonia: risk factors. AB - Postoperative pulmonary complications frequently lead to increased patient morbidity and mortality, hospital length of stay, and resource utilization. Atelectasis and infectious complications account for the majority of reported pulmonary complications. Risk factors are thought to exaggerate pulmonary function deterioration, which occurs both during and after surgical procedures. This article reviews the literature and describes risk factors frequently identified in relation to pre-, intra-, and postoperative settings, impact of each risk factor on pulmonary function, and issues related to risk factor evaluation. Eighteen risk factors are reviewed regarding their pathophysiologic impact on pre-, intra-, and postoperative pulmonary function. Key issues related to risk factor evaluation are also discussed. Identification of risk factors and prediction of postoperative pulmonary complications are important. Early identification of patients at risk for postoperative pulmonary complications can guide our respiratory care to prevent or minimize these complications. PMID- 7489037 TI - The effect of thoracic surgery on pulmonary function. AB - BACKGROUND: Pulmonary function is a main factor influencing postoperative morbidity and mortality in thoracic surgery. OBJECTIVE: To determine the significance of surgical trauma on postoperative pulmonary function by comparing the preoperative forced expiratory volume in 1 second with that obtained 14 days after surgery. METHODS: A retrospective study was conducted on 385 consecutive patients undergoing elective thoracic surgery over 3 years. RESULTS: Operative trauma alone effected a reduction of approximately 0.6 L per second in the value of forced expiratory volume in 1 second irrespective of the extent of pulmonary resection. Significant differences in the overall reduction among enucleation, segmental resection, lobectomy, and pneumonectomy were not observed. CONCLUSIONS: The data indicated that the remaining lung tissue was severely compromised throughout the postoperative period and that surgical trauma was the main factor influencing postoperative pulmonary function for at least 2 weeks. PMID- 7489038 TI - Use of a clinical assessment tool for orally intubated patients. AB - BACKGROUND: Although oral hygiene is difficult to perform in orally intubated, critically ill patients, oropharyngeal status may be related to acquisition of nosocomial organisms and should therefore be addressed. OBJECTIVE: To develop a clinical tool for assessment of the mouth in orally intubated patients. METHODS: Sixteen orally intubated patients from one surgical and one medical intensive care unit were subjects. Demographic, clinical, and baseline oropharyngeal assessment data, including sputum and mouth cultures, were obtained on patient admission to the study and every other day. RESULTS: Oropharyngeal cultures of four subjects grew nosocomial bacterial organisms and of two subjects, fungal species. These same organisms were cultured from sputum specimens. Seven patients displayed severe xerostomia; 10 lip lesions were identified in nine subjects, 8 tongue lesions in nine subjects, and 8 mucosal lesions in nine subjects. CONCLUSION: Systemic oropharyngeal assessment in critically ill orally intubated patients may prevent more serious oropharyngeal infections. PMID- 7489039 TI - Pressure ulcer risk factors in an ICU population. AB - BACKGROUND: Although it is well known that pressure ulcers are associated with negative patient outcomes and increased hospital cost, there is little research related to pressure ulcers in an intensive care unit population. OBJECTIVE: To determine the relative contribution of risk factors in the development of pressure ulcers in intensive care unit patients. METHOD: In an exploratory descriptive design, a convenience sample of 85 adults was used. Patients were enrolled in the study within 24 hours of admission to the intensive care unit; data were collected every other day until discharge from the intensive care unit. Instruments included a demographic data form, Braden Scale for Predicting Pressure Sore Risk, Skin Assessment Tool, and Decubitus Ulcer Potential Analyzer. RESULTS: The most common reasons for admission to the intensive care unit included multiple trauma from motor vehicle accidents, gunshot and stab wounds, and gastrointestinal bleeding. A pressure ulcer developed in 48 subjects. There were no significant differences in age, gender, history of diabetes or smoking, or medical diagnoses between patients in whom a pressure ulcer developed and those in whom it did not. Data analysis indicated that a Braden Scale score of 11, rather than the recommended score of 16, was statistically significant for predicting pressure ulcer risk. CONCLUSIONS: The results suggest that a cut-off score on the Braden Scale could be specific to an intensive care unit trauma population. PMID- 7489040 TI - Assessing patient comfort: local infiltration of lidocaine during femoral sheath removal. AB - BACKGROUND: Patient comfort is a primary nursing concern when removing femoral sheaths. The use of lidocaine infiltration, a common practice at our facility, was not addressed in the reviewed literature. OBJECTIVE: To assess and evaluate the level of patient comfort and efficacy of lidocaine hydrochloride as a local anesthetic during femoral sheath removal and application of a compression device. METHODS: A literature search and telephone interviews were conducted with nurses and technicians who removed or assisted with the removal of femoral sheaths in 13 hospitals in the United States. Three patient comfort indicators were then developed and used for data collection. Over a 12-month period, nurses in the cardiology interventional unit removed the femoral sheaths without the use of lidocaine from 111 angioplasty/arthrectomy patients and evaluated the level of patient comfort. RESULTS: A literature search provided no information that supported the use of lidocaine hydrochloride infiltration during sheath removal. Telephone interviews showed that six of the hospitals used lidocaine as standard procedure and seven did not use lidocaine at all. Of the evaluated patients, 105 (94.59%) reported comfort during sheath removal without local infiltration of lidocaine. CONCLUSION: Our results supported the premise that patients do not require lidocaine before sheath removal to maintain an acceptable level of comfort. PMID- 7489041 TI - Risk factors of adolescent and young adult trauma victims. AB - BACKGROUND: Repeated injury, or recidivism, because of intentional or unintentional injury is a growing chronic health problem among urban adolescents and young adults in the United States. OBJECTIVE: To describe demographic, social, environmental, psychological, and developmental antecedents and risk taking behaviors, and to examine their relationships to type of trauma and rate of trauma recidivism in adolescent and young adults in an urban trauma center. METHODS: One hundred adolescent and young adult trauma victims in an urban trauma center were interviewed, using the Adolescent Risk-Taking Instrument, the Brief Anger/Aggression Questionnaire, and the Trauma Risk Factor Interview Schedule. Bivariate correlation, multiple regression, and discriminant function analysis were used to examine the data. RESULTS: Of the sample, 89% experienced trauma related to interpersonal violence, including firearm injuries, stab wounds, and blunt trauma. Male gender, unemployment, past arrest, lower levels of spirituality, and higher levels of anger/aggression and thrill-seeking accounted for 25% of the variance in the number of risk-taking behaviors. Factors such as male gender, past arrest, unemployment, having been a crime victim in the past, lower autonomy, use of weapons, fighting, and no psychological counseling distinguished subjects with firearm-related injuries from subjects with other injury sources. Use of alcohol on weekdays, past arrest, and higher education levels were associated with trauma recidivism, explaining 14% of the variance. CONCLUSIONS: Social/environmental and psychological/developmental variables, as well as risk-taking behaviors, are important correlates of trauma and recidivism. These findings suggest the importance of advocacy for social policies conductive to reducing the risks of violence and trauma and risk-reduction interventions as components of posttrauma care. PMID- 7489042 TI - Suburban geriatric trauma: the experiences of a level I trauma center. AB - BACKGROUND: As the mean age of the US population increases, so does the incidence of geriatric trauma. Investigators have shown that the elderly have high morbidity and mortality rates associated with traumatic injuries. OBJECTIVE: To compare the severity of injury, mortality, and functional outcomes of geriatric patients with younger patients admitted to a suburban trauma center. METHOD: A convenience sample of trauma patients who were 65 years old or older was compared with trauma patients who were 35 to 45 and 55 to 64 years old. Demographic data, injury data, Injury Severity Scores, Revised Trauma Scores, length of stay, and functional ability outcomes were abstracted from a trauma registry in aggregate form and then analyzed. RESULTS: The sample consisted of 766 subjects (age 35-45, n = 223; age 55-64, n = 135; age 65 and older, n = 408) with a mean age of 64.6 years. A larger percentage of the elderly were victims of falls; younger trauma patients were more likely to be victims of motor vehicle crashes. Significant differences were found between age groups on Glasgow Coma Scale scores. Revised Trauma Scores, and length of stay. Significant differences were not found on Injury Severity Scores, mortality rates, or functional outcomes. CONCLUSIONS: Although anatomic injury severity of elderly patients was similar to that of younger patients, the elderly demonstrated greater physiologic compromise and longer hospital stays. Mortality rates were lower for the elderly group, but this result might be because a larger proportion of elderly patients were hospitalized with minor or moderate injuries. PMID- 7489043 TI - Presumed consent: the solution to the critical organ donor shortage? AB - Extraordinary advances in science and medicine such as transplantation of human tissues not only bring about previously unimaginable societal benefits but also create profound implications that involve autonomy and belonging, opposing moral considerations, and legal concerns. Today, technology is changing faster than our values. The issue of salvaging organs from the dead to meet the escalating need for human organs for lifesaving organ transplantation has evolved into an intricate web of interdisciplinary concerns and value conflicts; right and wrong are opinions and consensus does not seem to exist. This organ supply-demand mismatch, as well as suggestions for its resolution, has become a major challenge to the transplant community and to those in political and bioethical arenas. A methodical transition to presumed consent, or opting-out legislation, which removes the burden of decision from the family and the burden of request from the healthcare professional, may be the only solution to correct the imbalance between human organ need and availability. PMID- 7489044 TI - Impact of aborted sudden cardiac death on survivors and their spouses: the phenomenon of different reference points. AB - BACKGROUND: Although anecdotal data suggest that spouses of aborted sudden cardiac death survivors become enmeshed in the physical and emotional recovery of their mates, few longitudinal studies address the personal struggle of aborted sudden cardiac death survivors and their spouses during recovery. OBJECTIVE: To identify and explore phenomena experienced by aborted sudden cardiac death survivors and their spouses, and to determine implications for care. METHODS: This was a phenomenological study; qualitative interviews were conducted within 3 weeks of aborted sudden cardiac death and continued through 24 weeks after arrest at 6- to 8-week intervals. A total of 180 interviews were conducted with 40 survivors and 30 spouses. Interviews were tape-recorded, transcribed, and analyzed for recurrent themes. RESULTS: For the spouses the point of focus, or reference point, for future decision making was the arrest; for the survivors the reference point was prearrest life. These different reference points led to different concerns between spouses and survivors, from which spousal protectiveness and entrapment emerged. CONCLUSIONS: Acknowledgment of different reference points is essential in planning interventions for aborted sudden cardiac death survivors and their spouses. This population must be encouraged to express their questions, concerns, and fears early. Differences in perspectives should be identified to avoid troubled communication and conflicts. PMID- 7489045 TI - Antitachycardia pacing: an overview. AB - Ventricular tachycardia is the most common life-threatening tachyarrhythmia seen in patients with structural heart disease. In the past 10 years, many thousands of lives have been saved by the addition of the implantable cardioverter defibrillator to the armamentarium of treatment options. Yet, despite the success of these devices in the prevention of sudden cardiac death, many patients felt that the psychological cost was too high. Loss or restriction of some of the most basic activities such as driving and working and not knowing when they would receive the life-saving but painful shock took a toll on patients' perceptions of the quality of their lives. When antitachycardia pacing was demonstrated to provide about half of these patients a life-saving, yet comfortable, means of controlling episodes of ventricular tachycardia, physicians and patients were eager to try it. None of the many ways to provide antitachycardia pacing has proved overwhelmingly more effective than the others. This paper describes and illustrates each type of antitachycardia pacing and demonstrates their programmed parameters. Several clinical case illustrations are included. PMID- 7489047 TI - Clinical diagnosis of chest pain. PMID- 7489046 TI - The long QT syndrome: when syncope is common to the young and the elderly. PMID- 7489048 TI - No universal signs for acute myocardial infarction. PMID- 7489049 TI - Withdrawal of ventilator support. PMID- 7489050 TI - Advantages for the use of the activated clotting time (ACT) PMID- 7489051 TI - Exposure assessment in industry specific retrospective occupational epidemiology studies. AB - Quantitative estimation of exposure for occupational epidemiology studies has received increasing attention in recent years and, as a result, a body of methodological literature has begun to take form. This paper reviews the generic issues in the methodology of exposure assessment, particularly methods for quantitative retrospective assessment studies. A simple framework, termed an exposure data matrix (EDM), for defining and analysing exposure data is proposed and discussed in terms of the definition of matrix dimensions and scales. Several methods for estimation, interpolation, and extrapolation, ranging from subjective ratings to quantitative statistical modelling are presented and discussed. The various approaches to exposure assessment based on the EDM concept are illustrated with studies of lung disease among coal miners and other dust and chemically induced chronic occupational diseases. The advantages of validated statistical models are emphasised. The importance of analysis and control of errors in exposure assessments, and integration of the exposure assessment and exposure-response processes, especially for emerging occupational health issues, is emphasised. PMID- 7489052 TI - Occupational exposures estimated by means of job exposure matrices in relation to lung function in the PAARC survey. AB - OBJECTIVES: The aim of this analysis of the French Cooperative PAARC (Pollution Atmospherique et Affections Respiratoires Chroniques) survey, was to test whether occupational exposures to dusts, gases, or chemical fumes or to specific hazards, estimated by job exposure matrices, were related to a decrease in forced expiratory volume in one second (FEV1). METHODS: The most recent occupation was recorded in adults, aged 25-59, from non-manual worker households. Analysis was restricted to 10,046 subjects whose occupation was encountered at least 10 times in the study and who performed good FEV1 tracings. From occupational title, exposures to dusts, gases, and chemical fumes, and to specific hazards were classified in three categories (no, low, and high) with a British, a French, and an Italian job exposure matrix. Specific hazards were analysed for the British and French job exposure matrices for the same 42 specific dusts, gases, and chemical fumes. To limit spurious associations, a selection of seven hazard groups and 12 specific hazards was set before the start of the analysis. Based on the consistency of the relations according to sex and the British and French job exposure matrices, associations of age, height, city, and smoking adjusted FEV1 score with occupational exposures were classified as very likely, possible, or unlikely. RESULTS: For the three job exposure matrices and both sexes clear exposure-response relations between the level of exposure to dusts, gases, and chemical fumes, and a decrease in FEV1 were found. Associations with FEV1 were classified as very likely for known hazards such as organic dusts and textile dusts, and not previously recognised hazards such as polycyclic aromatic hydrocarbons (PAHs) and detergents, and as possible for solvents, waxes and polishes, and diesel fumes. Associations found for PAHs and solvents were confirmed by the Italian job exposure matrix. Associations remained significant in women, but not in men, after adjustment for educational level. CONCLUSIONS: Hypotheses have been generated for exposure to detergents, PAHs, and solvents, but they need to be interpreted with caution before replication. Significant associations found for known risk factors with a decrease in FEV1 are arguments for the validity of the matrices. Despite the expected limitations of job exposure matrices, these results encourage further work to improve exposure assessment by job exposure matrices. PMID- 7489053 TI - The risk of acquiring Q fever on farms: a seroepidemiological study. AB - OBJECTIVES: To determine the occupational risk of Q fever. DESIGN: Cohort study. SETTING: Community: five English local authority districts. SUBJECTS AND METHODS: Prevalence and incidence of immunoglobulin G (IgG) specific antibody to Coxiella burnetii phase II antigen was measured in a representative (study) cohort of farm workers in the United Kingdom, and detailed exposure data were collected. Also seroprevalence of Q fever in a (control) cohort of police and emergency service personnel was measured. RESULTS: Prevalence was significantly (P < 0.01) higher in the study cohort (105/385 v 43/395). During the first 12 month period after enrollment no seroconversions were found (upper 95% confidence limit: 1318/100,000/year). During the second 12 month period after enrollment two seroconversions were found, equalling an incidence of 813/100,000/year (95% confidence interval (95% CI) 98-2937/100,000/year). No association was found between seroprevalence and age. In the study cohort, extent of total contact with farm animals seemed more important than exposure to any specific animal: full time employees were more than four times more likely to be antibody positive than part time employees (P < 0.05). Exposure to cattle, but not sheep, goats, cats, raw milk, and hay (all reported sources of Q fever) was associated with being positive to Coxiella burnetii IgG by univariate analysis but this association was not independent of total farm animal contact. CONCLUSIONS: The risk of Q fever on livestock farms is related to contact with the farm environment rather than any specific animal exposure. The absence of an increasing prevalence with age suggests that exposure may occur as clusters in space and time (outbreaks). PMID- 7489054 TI - Neurotoxicity among pesticide applicators exposed to organophosphates. AB - OBJECTIVES: An epidemiological study of 90 male pesticide applicators licensed in New York was conducted to investigate the effect of exposure to organophosphate pesticides on the peripheral nervous system. METHODS: A cohort of farmers and pesticide applicators from New York State were questioned off season (November 1988-February 1989) and again during the spraying season (April 1989-August 1989) about the presence of several acute signs and symptoms. Short term exposure was validated by measuring the concentration of dimethylthiophosphate (DMTP), a metabolite of guthion, in urine. Chronic signs of subtle peripheral nerve damage were determined by vibration threshold sensitivity of the farmers and applicators tested during November 1988-February 1989 and compared with controls drawn from the general population who were tested during the same time period the next year (November 1989-February 1990). Vibration threshold sensitivity was determined for both the hands and feet. Long term exposure to pesticides was determined by questionnaire. RESULTS: Paired t tests show that mean vibration threshold scores were significantly higher for the dominant (P < 0.00) and non-dominant (P < 0.04) hands among pesticide applicators when compared with scores for population based controls individually matched on age, sex, and county of residence. CONCLUSIONS: A significant increase in mean vibration threshold sensitivity for the dominant and non-dominant hand suggests previous organophosphate exposure among pesticide applicators was associated with a loss of peripheral nerve function. PMID- 7489056 TI - Platinum salt sensitivity in refinery workers: incidence and effects of smoking and exposure. AB - OBJECTIVE: To measure the incidence of platinum salt sensitivity (PSS) in refinery workers and examine the influence of cigarette smoking and exposure to platinum salts on sensitisation. DESIGN: A prospective cohort study with examination of workers at quarterly intervals for 18 months, and again at 24 months. SETTING: A South African primary platinum refinery. SUBJECTS: 78 new recruits, selected by the refinery's usual procedure, without apparent atopy and in good respiratory health. RESULTS: After 24 months 32 (41%) subjects had been diagnosed PSS and were subsequently medically separated. Twenty two (28%) cases were confirmed by positive skin prick test to platinum salts, 10 (13%) cases were symptomatic but skin prick negative. Incidence of cases per 100 person-months was 1.9 skin prick positive and 0.8 negative. Risk of sensitisation was about eight times greater for smokers than non-smokers, and six times greater for high exposure than low exposure. CONCLUSION: Smoking and intensity of exposure were definitely associated with development of PSS. Positive responses to platinum salt skin prick test had a 100% positive predictive value for symptoms and signs of PSS if exposure continued. PMID- 7489055 TI - Lung function and chronic respiratory symptoms of pig farmers: focus on exposure to endotoxins and ammonia and use of disinfectants. AB - OBJECTIVES: The prevalence of chronic respiratory symptoms among pig farmers is known to be high, but the aetiology of these symptoms is not yet unravelled. Long term average exposure to dust was evaluated, endotoxins and ammonia and use of disinfectants were compared with chronic respiratory symptoms and depressed base line lung function. METHODS: A cross sectional study was performed among 194 Dutch pig farmers, of whom 100 had not and 94 had chronic respiratory symptoms. Exposure-response relations were evaluated with multiple logistic and linear regression analysis. Estimates of long term average exposure were based on two personal exposure measurements, taken on one day in summer and one day in winter. Information on use of disinfectants and disinfection procedures was assessed by a walk through survey and interview by telephone. RESULTS: Exposure to dust, endotoxins, and ammonia were not related to chronic respiratory symptoms. Duration of the disinfection procedure and pressure used at disinfection were strongly and positively related to chronic respiratory symptoms. A significant inverse association between base line lung function and endotoxin exposure was found only among asymptomatic farmers. Ammonia exposure and duration of the disinfection procedure were significantly associated with base line lung function in the entire population. CONCLUSION: Results suggest that use of disinfectants is an important aetiological factor in chronic respiratory health effects of pig farmers. This factor has not been studied before. Results also suggest an aetiological role for exposure to endotoxins and ammonia in development of chronic respiratory health effects, but longitudinal studies with detailed exposure assessment strategies are required to assess their roles. PMID- 7489057 TI - Mortality and cancer incidence among secondary lead smelter workers. AB - OBJECTIVES: To examine the mortality pattern and the cancer incidence in a cohort of long term smelter workers exposed to lead. METHODS: The cohort consists of 664 male lead battery workers, employed for at least three months in 1942-87. From 1969 the values of all blood lead samples repeatedly obtained from these workers every two to three months, have been collected in a database. The expected mortality and morbidity 1969-89 was estimated from the county rates, specified for cause, sex, five-year age groups, and calendar year. Individual exposure matrices have been calculated and used for dose-response analyses. RESULTS: The total cohort showed an increased overall mortality (standardised mortality ratio (SMR) 1.44; 95% confidence interval (95% CI) 1.16-1.79), an increased mortality from ischaemic heart diseases (SMR 1.72; 95% CI 1.20-2.42) and all malignant neoplasms (SMR 1.65; 95% CI 1.09-2.44). These risk estimates were unaffected or slightly decreased when applying a latency period of 15 years, and no dose response pattern was shown. The non-significantly raised cancer incidence in the gastrointestinal tract (11 malignancies) in the total cohort, increased to a barely significant level in the quartile with the highest cumulative lead exposure (standardised incidence ratio (SIR) 2.34, 95% CI 1.07-4.45). No clear dose response pattern was evident when further subdividing the data into those first employed up to 1969 v those first employed after 1969 when the blood lead monitoring programme started. The risk estimate for malignancies in the gastrointestinal tract was not related to latency time. The cancer incidence was not increased at other sites. CONCLUSIONS: A slightly increased incidence of gastrointestinal cancers was found in workers exposed to lead and employed before 1970. The lead cohort also showed an increased mortality from ischaemic heart diseases. These risk estimates did not show a dose-response pattern and were not associated with latency time. The results must also be interpreted with caution because of limited numbers, and lack of dietary and smoking data. PMID- 7489060 TI - Reports of work related musculoskeletal injury among home care service workers compared with nursery school workers and the general population of employed women in Sweden. AB - OBJECTIVES: To describe the nationwide occurrence of work related musculoskeletal injuries among all home care service workers in Sweden, and to identify relative risks and risk factors of the injuries. METHODS: The study was based on work related injuries reported to the Swedish occupational injury information system in 1990-1. The work related musculoskeletal injuries were divided into overexertion accidents and musculoskeletal diseases. The incidence of the injuries in female home care service workers was compared with those in nursery school workers and all other employed women in Sweden. RESULTS: In home care service workers, the annual incidence of injury from overexertion accidents and musculoskeletal diseases were 19.2 and 15.1 per 1000 workers, respectively, which was higher than those in nursery school workers and all employed women in Sweden. For five injury locations including the back, all the age standardised relative risks (SRR) of overexertion accidents exceeded 4.0, and most of those for musculoskeletal diseases were 1.5 or more in home care service workers compared with all other employed women in Sweden. Total duration of sick leave due to overexertion accidents was 7.7 times, and musculoskeletal diseases 3.5 times, longer than in nursery school workers. National loss due to sick leave resulting from only musculoskeletal injuries in home care service workers was about 8.2% of the total work related sick leave in all employed women in Sweden, although the number of home care service workers represented only some 5% of this population. Lifting other people was most frequently reported as the main risk cause of overexertion accidents in both kinds of workers. CONCLUSIONS: The results support the hypothesis that home care service workers have higher annual injury incidence of musculoskeletal injuries than nursery school workers due to physically stressful tasks that are far less common in nursery school workers. PMID- 7489061 TI - Thetford plastics fire, October 1991: the role of a preventive medical team in chemical incidents. AB - OBJECTIVES: To review the role of a medical team in the emergency management of a major polyvinyl chloride (PVC) fire in an urban area. METHODS: The district health authority's consultant in communicable disease control (CCDC) was requested to advise on the health impacts of a fire that consumed some 1000 tonnes of plastic, mainly PVC, over 72 hours and which emitted a large smoke plume that threatened the health of local residents and emergency workers alike, constituting one of the largest incidents the local emergency services had dealt with in recent years. A medical team was formed comprising the CCDC, a regional epidemiologist, an occupational physician, and a medical toxicologist. This paper is an account of this team's experience of advising on the medical management of the emergency without having any formally established role or previous training for the task. RESULTS: The main issues requiring the input of the medical team included: the possible products of combustion and their effects on health; the clinical management of those exposed; the alerting of local hospitals to the type of casualties to expect; the special health risks posed to emergency workers, especially the firemen; the need for evacuation of local residents; the risks of contamination of soil, water, and crops; the potential health impact of the plume; and the provision of expert and authoritative advice on the short and long term health implications to the public. Active surveillance systems, which included the local general practitioners and hospitals, were established and air monitoring instigated. The 46 casualties were restricted to emergency personnel who had inadvertently received exposure to the fire smoke: all recovered within 48 hours. Local residents were unharmed. CONCLUSION: The incident showed the need for preventive medical teams trained to fill a formal advisory and investigative role for chemical releases and fires, and which can play an integral part in emergency management. PMID- 7489058 TI - Haematopoietic cancer mortality among vehicle mechanics. AB - OBJECTIVE AND METHODS: This historical cohort study investigated causes of death among car and mobile equipment mechanics in the District of Columbia's Department of Public Works. Men who were employed for at least one year between 1977 and 1989 were eligible for inclusion in the cohort; follow up was up to the end of 1991. Three cases of leukaemia (index cases) had been reported among these workers before the inception of this study. This research was undertaken to estimate the relative risk of haematological cancer among mechanics working for the District of Columbia. RESULTS: Among the 335 male fleet maintenance workers, the all cause standardised mortality ratio (SMR) was 0.50 (33 observed deaths, 95% confidence interval (95% CI) 0.35-0.70), and the all cancer SMR was 0.55 (nine deaths, 95% CI 0.25-1.05). Three deaths from lymphatic and haematopoietic cancer were observed; the SMR was 3.63 (95% CI 0.75-10.63). In the subgroup with highest potential for exposure to fuels and solvents, the SMR for leukaemia and aleukaemia was 9.26 (two deaths, 95% CI 1.12-33.43), and the SMR for other lymphatic and haematopoietic neoplasms was 2.57 (one death from malignant lymphoma, 95% CI 0.06-14.27). All three lymphatic and haematopoietic cancer deaths were among car and mobile equipment mechanics (one was an index case). The two additional index cases were a fourth mechanic who died of leukaemia in 1992, after mortality follow up ended, and a fifth mechanic who was diagnosed with leukaemia in 1988 and is still alive. CONCLUSION: Many garage mechanics in this cohort regularly used petrol to clean parts and to wash their hands; some workers would occasionally siphon petrol by mouth. Benzene, a recognised cause of haematological cancer, is a component of petrol. Previous research indicates that garage mechanics may be at risk of leukaemia and other haematological cancers, presumably due to exposure to petrol; this study supports those findings. PMID- 7489059 TI - Hepatic ultrasonic changes in workers exposed to perchloroethylene. AB - OBJECTIVE: To determine if subclinical hepatotoxicity is associated with exposure to perchloroethylene at concentrations commonly experienced in the workplace, and whether surveillance with serum hepatic transaminase activity underestimates such effects. METHODS: Hepatic parenchymal echogenicity on ultrasonography and serum hepatic transaminase activity were compared in 29 community based dry cleaning operators exposed to perchloroethylene, and a control group of 29 non-exposed laundry workers. Perchloroethylene exposure was assessed by work history and air monitoring. RESULTS: Mean hepatic transaminase activities were minimally increased in dry cleaners compared with laundry workers. Increased alanine aminotransferase activities, between 1.0 and 1.5 times the normal limits, were found in five of 27 (19%) dry cleaners compared with one of 26 (4%) laundry workers. In contrast, diffuse parenchymal changes in echogenicity, as determined by hepatic ultrasonography, were increased nearly twofold in dry cleaners, occurring in 18 of 27 (67%) dry cleaners compared with 10 of 26 (39%) laundry workers (P < 0.05), and were most strongly associated with increased perchloroethylene exposure in older dry to dry or wet transfer operations (odds ratio 4.2, 95% confidence interval 1.1-15.3). Mean eight hour time weighted average perchloroethylene exposure for dry cleaners was 16 ppm, which is less than the permissible exposure limit of 100 ppm in the United States. CONCLUSIONS: It was concluded that mild to moderate hepatic parenchymal changes occur more frequently in workers exposed to perchloroethylene than in populations not exposed to chemical solvents, and that these effects are underestimated by serum hepatic transaminase activity. PMID- 7489063 TI - Back pain and male parenthood. PMID- 7489062 TI - Does occupational exposure to dust prevent colorectal cancer? PMID- 7489064 TI - Preliminary experimental findings using intraperitoneal assays to determine carcinogenic potential of man made mineral fibres: relevance to recent proposals for classification testing. PMID- 7489065 TI - A study of chromium induced allergic contact dermatitis with 54 volunteers: implications for environmental risk assessment. PMID- 7489066 TI - Changes in survival patterns of very low-birth-weight infants from 1980 to 1993. AB - OBJECTIVE: To determine changes in survival patterns among very low-birth-weight ( < 1500 g) infants between 1980 and 1993. METHODS: The records of 12,960 infants treated in nine perinatal intensive care centers in Florida were analyzed on the basis of survival (discharged alive from hospital) according to four independent variables: birth weight, race, sex, and transport status. Survival curves were generated using log linear regression techniques for each race by sex by transport status group. RESULTS: Race, sex, and transport status correlated significantly with survival: survival percentages were higher among black infants, female infants, and infants transported to the perinatal intensive care centers than among white infants, male infants, and those admitted initially to the tertiary care centers. After 1985, 95% of neonates with birth weights between 1200 and 1500 g survived. In addition, survival of 500- to 500-g transported black male infants increased from zero to near 80% during the 13-year period; that of 500- to 550-g inborn white female infants rose from 35% to 70%. CONCLUSIONS: These results illustrate the value of taking into account race, sex, and transport status in efforts to understand the contribution that neonatal intensive care of extremely low-birth-weight infants makes to the lowering of infant mortality, and of using multivariable statistical procedures to generate predicted survival probabilities for different subpopulations. These probabilities can be applied to (1) predicting survival for specific subgroups of extremely low-birth-weight infants, and (2) helping physicians develop clinical guidelines for extending care to infants at the threshold of viability. PMID- 7489067 TI - Children's and women's ability to fire handguns. The Pediatric Practice Research Group. AB - OBJECTIVES: To evaluate whether strength differences between children and women might keep children from firing handguns and to determine how many young children can fire available handguns. DESIGN: One- and two-index finger trigger-pull strength was tested using a standard protocol. Data on trigger-pull settings of 64 commercially available handguns were obtained. SETTING AND PARTICIPANTS: Convenience sample of well children and their mothers at four Chicago (Ill)-area pediatric practices for health supervision visits, and of siblings of emergency department patients, during an 8-week period. INTERVENTION: None. MAIN OUTCOME MEASURE: One- and two-index finger trigger-pull strength of mothers and children. RESULTS: Twenty-five percent of 3- to 4-year-olds, 70% of 5- to 6-year-olds, and 90% of 7- to 8-year-olds have a two-finger trigger-pull strength of at least 10 lb, the fifth percentile one-finger trigger-pull strength of adult women. Forty (62.5%) of 64 handguns require trigger-pull strength of less than 5 lb; 19 (30%) of 64 require 5 to 10 lb. CONCLUSIONS: Significant overlap exists in the trigger pull strength of young children and women, limiting the potential use of increased trigger-pull settings to discourage firearm discharge by children. Young children are strong enough to fire many handguns now in circulation. PMID- 7489068 TI - Correlates of multigravida women's binge drinking during pregnancy. A longitudinal study. AB - OBJECTIVE: To refine our understanding of prenatal psychosocial factors associated with binge drinking during pregnancy and the contribution of binge drinking during pregnancy to the duration of newborn hospitalization. DESIGN: Prospective cohort. SETTING: A large urban medical center in Wisconsin. PARTICIPANTS: One hundred thirty-nine women (106 multigravida) who were invited to participate during a prenatal clinic visit early in their third trimester. MEASURES: Prenatal measures included social support (Maternal Social Support Index), depression (Center for Epidemiologic Studies Depression Scale), stress (Difficult Life Circumstances), substance use (Monitoring the Future Substance Use Questionnaire and T-ACE Scale [a screen with questions about tolerance, annoyance, cutting down, and using alcohol as an eye-opener), and maternal-fetal attachment (Maternal-Fetal Attachment Scale). After delivery, a mothers' and infants' medical record review form was used. RESULTS: Multigravida pregnant women (n = 106) were older than primigravida pregnant women (n = 33) (25.8 +/- 0.6 vs. 20.5 +/- 0.5 years; P = .001), with more children at home (2.3 +/- 0.2 vs 1.3 +/- 0.3; P = .01) and less social support (Maternal Social Support Index, 20.1 +/- 0.6 vs 22.9 +/- 1.0; P = .03). All of the binge-drinking women in this sample were in the unmarried multigravida subgroup (17/101 [17%]). Compared with multigravida pregnant women who did not binge drink during pregnancy, binge drinking pregnant women were older (28.1 +/- 1.3 vs 25.1 +/- 0.6 years; P = .03) and more socially isolated (Maternal Social Support Index, 17.2 +/- 1.3 vs 20.7 +/- 0.7; P = .04) and were more likely to smoke during the pregnancy (82% vs 39%; P = .001). Even after controlling for a number of other important biologic and psychosocial factors (duration of pregnancy, maternal gravidity, racial heritage, education, social support at second trimester, and birth weight), by hierarchical multiple linear regression, binge drinking within the last 2 weeks before the late second-trimester interview continued to explain a significant amount of variance in duration of newborn hospitalization (total R2 = .48, partial R2 = .04; P = .01). CONCLUSIONS: This study suggests that binge drinking during pregnancy is related to longer newborn hospitalizations. Effective prenatal interventions to improve the outcome of pregnancies for women who abuse alcohol during pregnancy should use early screening and provide augmentation of mothers' social support. PMID- 7489069 TI - Body dissatisfaction and unhealthy weight-control practices among adolescents with and without chronic illness: a population-based study. AB - OBJECTIVE: To compare body dissatisfaction and unhealthy weight-loss practices among adolescents with and without chronic illness. DESIGN: Survey. PARTICIPANTS: The sample consisted of 2149 adolescent boys and girls with diabetes, asthma, attention deficit disorder, physical disabilities, or seizure disorders; and a comparison group of 1381 adolescents without chronic illness. MAIN OUTCOME MEASURES: Body concerns, binge eating, frequent dieting, vomiting, and laxative or diuretic use among adolescents. RESULTS: Adolescents with chronic illness reported higher body dissatisfaction and engaged in more high-risk weight-loss practices than adolescents without chronic illness. This trend was consistent across the different conditions and was not limited to those with a nutrition related condition such as diabetes. The trend remained after conducting logistic regression and controlling for age, race, socioeconomic status, and body mass index. CONCLUSIONS: Adolescents with chronic illness are at high risk for engaging in unhealthy weight-loss practices and should be screened and targeted for prevention and treatment. PMID- 7489070 TI - Topical iodine and neonatal hypothyroidism. AB - OBJECTIVES: To determine whether skin care practices with iodine-containing disinfectants are putting patients in the neonatal intensive care unit at risk for primary hypothyroidism. Cutaneous exposure to povidone-iodine antiseptic solutions may be a cause of primary hypothyroidism in neonates. DESIGN: Prospective pilot study. SETTING: Level III neonatal intensive care unit of a university-affiliated hospital. PARTICIPANTS: Sequential sample of 47 medial and surgical patients admitted to the neonatal intensive care unit who received cutaneous povidone-iodine applications in preparation for invasive or surgical procedures. METHODS: Seven to 10 days after iodine exposure, capillary blood samples were obtained on filter paper blots for thyroid function testing and urine samples were collected to determine quantitative iodine concentrations. A plasma creatinine level was determined for each subject. RESULTS: A total of 47 patients were enrolled. The gestational ages of subjects ranged from 26 to 41 weeks (mean, 33.6 weeks); the male-to-female ratio was 28:19; and the birth weights ranged from 0.7 to 5.1 kg(mean, 2.42 kg). The thyroxine level ranged from 20 to 187 nmol/L (1.6 to 14.6 micrograms/dL) (mean, 102 nmol/L [7.9 micrograms/dL];reference, > or = 90 nmol/L [> or = 7 micrograms/dL]; and the thyrotropin level ranged from 0.1 to 16.5 mU/L (mean, 6.4 mU/L; reference, < 20 mU/L). The mean uridine iodine concentration was 2798.0 micrograms/dL (reference, < 40 micrograms/dL), and the mean plasma creatinine level was 60 mumol/L (0.69 mg/dL) (reference, < or = 50 mumol/L [ < or = 0.6 mg/dL] for males and < or = 40 mumol/L [ < or = 0.5 mg/dL] for females). CONCLUSIONS: There was no documentation of primary hypothyroidism in our subjects despite elevated urine iodine levels. While it is still possible that patients who receive long-term iodine exposure in other settings (eg, cardiac catheterization) are at risk for primary hypothyroidism, our study suggests that the amount of iodine absorbed through routine neonatal intensive care unit procedures does not substantially alter thyroid function during the first 10 days of life. An important confounding variable is that seven patients were receiving dopamine hydrochloride infusions and four were receiving dexamethasone phosphate at the time of sample collection. We therefore cannot rule out the possibility that these medications masked a thyrotropin level elevation that would have occurred in a primary hypothyroid state. We discuss implications for the interpretation of the results of neonatal thyroid function tests. PMID- 7489071 TI - The undoing of a diagnosis. The effect of a misdiagnosis of a disease. AB - Eighteen children who had been diagnosed as having cystic fibrosis (CF) in earlier years were found to be normal and have been inappropriately diagnosed. Removal of the diagnosis from the child involved a movement through stages in the family from disbelief to anger before comfortable acceptance of normalcy in their child. Diagnosis of a chronic disease involves complex alterations in perception of self and family. The long-range effects of such an improper diagnosis may influence behavior and development. PMID- 7489072 TI - Prevalence of carrying a weapon and related behaviors in urban schoolchildren, 1989 to 1993. AB - OBJECTIVE: To characterize the prevalence of weapon carrying in an epidemiologic sample of about 1500 urban schoolchildren between spring 1989 and spring 1993. DESIGN: A descriptive analysis based on 5 years of cross-sectional confidential survey data. SETTING: An urban public school system in the mid-Atlantic region of the United States. PARTICIPANTS: The study base included all children who were interviewed in each year from 1989 to 1993, disregarding the number of prior or subsequent interviews. The mean age of the participants in the sample was 9 years when the survey began. MAIN OUTCOME MEASURES: Carrying a nonlethal or lethal weapon (eg, a stick, knife, or gun) to defend or to threaten or hurt someone, assessed by self-report. Reports of interpersonal aggression also were obtained. RESULTS: Knife and gun carrying increased with age while stick carrying decreased. In 1989, carrying a lethal weapon (eg, knife or gun) was reported by 11.7% of boys and 3.3% of girls; in 1993, these proportions had increased to 22.2% and 15.3%, respectively. Gun carrying increased over time; by 1993, 9.9% of boys and 1.4% of girls reported that they had carried a gun in the previous year. Carrying a weapon to defend or protect oneself was more commonly reported than to hurt or threaten someone. Various types of interpersonal aggression were frequently reported among boys and girls in each year. Our findings support the hypothesis that early involvement with a weapon is associated with later more serious involvement with a weapon. CONCLUSIONS: Involvement in weapons-related behavior starts young, well before the middle school year. Effective intervention programs may require attention to the early determinants of involvement with weapons, sex differences, and perhaps also the changing profile of reasons given for carrying and using weapons in conjunction with other forms of aggressive behaviors. PMID- 7489073 TI - Health status of preterm low-birth-weight infants. Comparison of maternal reports. AB - BACKGROUND: Developers of measures of child health status have documented acceptable reliability and some validity, but less attention has been paid to the concurrent and predictive validity of these measures. METHODS: We examined the concurrent and predictive validity of the RAND General Health Rating Index, the Stein-Jessop Functional Status II-R, and the mother's global assessment of her child's health on a 5-point scale, in a sample of preterm low-birth-weight children (n = 608) who were followed up as controls in the Infant Health and Development Program. We compared maternal-reported measures assessed at 24 months with other measures of growth, morbidity, functioning, and health care utilization assessed concurrently and at 36 months in bivariate and multivariate analyses. RESULTS: After controlling for other factors, the RAND General Health Rating Index and the Stein-Jessop Functional Status II-R were unrelated to the growth, utilization, or functioning measures. The RAND General Health Rating Index was significantly, but weakly, related to future morbidity. The mother's global perception of health was significantly related to outpatient utilization and behavior problems. CONCLUSIONS: Clinicians may find that maternal assessment of overall child health is a sensitive but nonspecific indicator of the mother's concern. For researchers, none of these measures seems likely to serve as a proxy for health care utilization or morbidity in studies of other phenomena, or as an indicator of detailed health outcomes. PMID- 7489074 TI - Longitudinal head growth in developmentally normal preterm infants. AB - OBJECTIVE: To determine growth in head circumference from birth to 18 months of age in normal infants with low birth weight. METHODS: Healthy, appropriate-for gestational-age, singleton, white infants weighing less than or equal to 2500 g at birth and with normal development at 18 months of age were included in this study. Serial measurements of head circumference (corrected for gestational age) from 450 eligible infants were compared with reference data for head circumference. RESULTS: Longitudinal measurements of head circumference for infants weighing more than 1000 g at birth were similar to reference data for term infants. Head measurements for infants weighing less than or equal to 1000 g at birth were notably smaller than the measurements in the reference data. A cubic spline curve drawn through the head circumference measurements between birth and age 18 months (corrected for gestational age) for infants weighing less than or equal to 1000 g at birth was significantly (P < .001) below the curve for infants weighing more than 1000 g at birth. At age 18 months, the mean difference in head circumference between the group weighing less than or equal to 1000 g at birth and the weights in the reference data was 1.6 cm (P < .01). (Data were analyzed with Wilcoxon's signed rank test.) CONCLUSIONS: These data show that head circumference grids are appropriate for observing head growth in infants with a birth weight more than 1000 g. However, head circumference growth for normal infants with birth weight less than or equal to 1000 g does not "catch up" with that of larger premature infants or term infants. PMID- 7489076 TI - Teaching residents about patient and practice termination in community-based continuity settings. AB - BACKGROUND: The pediatric residency program at the University of Massachusetts Medical Center, Worcester, has based its continuity experience in community practices since 1988. Residents develop a relationship not only with their patients but also with the preceptors, with whom they are paired one-on-one, and with office staff. OBJECTIVE: To describe the structure and results of an educational program that was developed to address the termination issues that arise at the end of residency. The educational program consists of four components: (1) a seminar, (2) a "mini-block" rotation, (3) office staff involvement, and (4) a resident-preceptor dinner. RESULTS: The following issues and themes have been recurrent in the discussions during the past 4 years: (1) the importance, for patient and resident, of identifying who will be the subsequent health care provider for the patient; (2) the inability to identify which patients had strongest attachment to residents; (3) parental surprise about the resident's departure, even though all parents had been told that the resident was going to be in the practice for only a limited period; (4) the desire of residents to have follow-up on patients after termination; (5) critical aspects of the process of informing patients about the resident's departure; (6) the importance of identifying and addressing the attachment of the resident to the preceptor and office staff, as well as to patients; and (7) the affirming experience that the termination sessions with the patients can be for the residents. CONCLUSION: Although the termination process is potentially emotionally difficult, it can be a personally and educationally valuable experience for residents. PMID- 7489077 TI - Medical informatics and pediatrics. Decision-support systems. AB - Decision support is an important area of medical informatics research. Computer based decision-support tools facilitate diagnosis and the management of patients after a diagnosis has been established. Diagnostic decision-support tools, such as Meditel, Quick Medical Reference, DXplain, Iliad, and PEM-DXP are potentially useful "expert systems." Other management-support tools, such as systems that use clinical practice guidelines to create reminders and alerts, also have been developed and evaluated. We do the following: (1) to provide an overview of diagnostic and management decision-support systems; (2) explore the background of and motivation behind these systems; (3) survey the uses of decision-support technology in office-based and inpatient pediatric practices; and (4) discuss the virtues and problems associated with some of these tools, and current controversies and future goals for computer-based decision support. PMID- 7489075 TI - Comparison of cocaine and opiate exposures between young urban and suburban children. AB - OBJECTIVE: To determine the prevalence of cocaine and opiate metabolites in the urine of young urban and suburban children. DESIGN: Survey. SETTING: Urban and suburban emergency departments and private pediatric practices. PATIENTS: A convenience sample of 1469 children between 1 and 60 months of age who required a urinalysis for investigation of the chief complaint. INTERVENTION: None. MAIN OUTCOME MEASURES: Urine was screened for benzoylecogonine and opiates using an enzyme-multiplied immunoassay technique and a fluorescence-polarization immunoassay, both with a sensitivity of 50 ng/mL. RESULTS: Benzoylecogonine was identified in the urine of 45 children (3.1%) (95% CI, 2.2% to 3.9%) and opiates in the urine of 38 children (2.6%) (95% CI, 1.8% to 3.4%). No difference was observed between urban and suburban health care facilities in the percentage of patients whose urine tested positive for benzoylecgonine (29 of 1011 vs 16 of 458, P = .6) or opiates (28 of 1011 vs 10 of 458, P = .6). CONCLUSION: Exposure to illicit drugs, as reflected by urinary metabolites, is similar for urban and suburban children. PMID- 7489078 TI - Policy and poverty. Child and community health in Philadelphia, 1900 to 1930. AB - On a fall day in 1913, a man sat on a crowded wooden bench with a little boy on his lap. He waited all day, only to be told that the physician who would treat his son was not working in the public clinic that day. The next morning he learned that if he returned at 3 o'clock in the afternoon, he would stand a better chance of seeing the physician. Since the family home was a shack and even food was scarce, he could ill afford the loss of another day's pay. His son had infantile paralysis, though, and the father believed that the physician could help, so he kept trying. The boy died during the Christmas holidays. "Is it simply that there are not enough public clinics, enough doctors willing to help care for the people who come to them?" asked the social worker who told this story. "Or is there something wrong with the system?" PMID- 7489079 TI - Radiological case of the month. Hepatic hemangioendothelioma. PMID- 7489080 TI - Picture of the month. Cleiodocranial dysplasia. PMID- 7489081 TI - Pathological case of the month. Wilms' tumor (nephroblastoma). PMID- 7489082 TI - The pediatrician and cancer prevention. PMID- 7489083 TI - Urine concentration and enuresis in healthy preschool children. PMID- 7489084 TI - Factors associated with prolonged hospitalization of children with asthma. PMID- 7489086 TI - [The concept of the biological action of low doses of ionizing radiation on cells]. AB - A concept of biological cellular effect of ionizing radiation at small doses was suggested on the basis of analysis of physical interactions of ionizing radiation with substance as well as of experimentally observed cell reactions to low dose radiation. Probable molecular-cellular mechanisms of cell response at low level irradiation were considered. PMID- 7489085 TI - [A critical analysis of the current concepts and approaches to assessing the biological action of low doses of ionizing radiation]. AB - Analysis of up-to-date conceptions and approaches to assessment of biological effect of small doses of radiation showed that presently there is no unified concept of biological effect of ionizing radiation in the range of low doses. The models using for genetic risk assessment are usually based on the linear non threshold concept and have insufficient biological justification. They are of extrapolation nature and come into conflict with available experimental data. The analysis of experimentally observed cell reactions to irradiation allows the author to conclude that the behavior of genetical disorders in the range of low doses in universal and could be approximated by linear-threshold curve. The doses at which the change of dependence takes place are different for various objects and determined by their sensitivity to external influence. PMID- 7489087 TI - [The results of the cytogenetic examination of children and adolescents living in radionuclide-contaminated regions of Kaluga Province]. AB - After the Chernobyl accident in a period from 1989 to 1992 blood samples were collected and analysed for chromosomal aberrations in lymphocytes from 490 children born and living continuously in two districts of Kaluga Province with radioactive contamination. The results of cytogenetic examinations show the frequency of unstable aberrations in children from Zhizdra District to be higher than that in children from Ulyanovo District and in the control group. No correlation was found between the thyroid absorbed doses ranging from 0 to 5 Gy and the aberration frequencies. No differences in chromosomal aberration yields were observed in children born before or after the Chernobyl accident. Repeat examinations of the same children cohorts revealed no significant changes with time in the level of unstable aberrations. PMID- 7489088 TI - [The cytogenetic effects in the population of the Altai Territory subjected to ionizing radiation exposure as a result of the nuclear explosions at the Semipalatinsk proving grounds]. AB - The cytogenetic examination of 178 persons living in seven settlements of the Altai region exposed to ionizing radiation during Semipalatinsk nuclear tests in 1949-1962 was carried out. It was shown that more than 80% of absorbed dose was due to the first nuclear explosion in 1949. The frequency of chromosome aberrations (dicentrics and rings) significantly exceeded the control level. The linear correlation between the dicentrics and rings frequency and absorbed dose was found. The cells with more than one aberrations were revealed in irradiated persons. This is supposed to be the result of 239Pu alpha-radiation. PMID- 7489089 TI - [The results of the cytogenetic examination of the inhabitants of population centers adjacent to the Semipalatinsk nuclear testing grounds (at the period of the work of the interdepartamental commission in 1989)]. AB - Within the frames of activity of joint commission the cytogenetic studies of 98 persons from different areas of Semipalatinsk Province have been carried out in 1989. The studies revealed the higher level of chromosome aberrations in the settlements adjacent to firing ground. The possible connection of discovered cytogenetic lesions to the influence of radiation aftermath of ground and air nuclear weapon test is discussed. PMID- 7489091 TI - [A comparative study of the frequency of stable and unstable chromosome aberrations in the gamma irradiation of human lymphocytes in vitro]. AB - Comparative studies of the yield of stable and unstable chromosome aberrations have been carried out in human lymphocytes gamma irradiated in vitro within the doze range 0.1--1.0 Gy. It was observed that the yield of unstable chromosome aberrations (dicentrics and centric rings) is satisfactorily described for discovered dose range with linear-quadratic approximation. For stable chromosome aberrations in G-banded chromosome the dose response dependence was less expressed. The studies revealed non-random involvement of different chromosomes in translocations, and the most frequently involved are 1 and 13 chromosomes. PMID- 7489092 TI - [The cytogenetic examination of different groups of children living in regions of Bryansk Province contaminated as a result of the Chernobyl accident]. AB - The frequency of chromosome aberrations (CA) was studied in peripheral blood lymphocytes from healthy children and children with thyreopathology living in the city of Klintsy, Bryansk Province (contamination level up to 5 Ci/km2), and two Moscow groups, respectively. We have observed the elevated number of dicentrics and acentric fragments in cell from children with thyreopathology with respect to those from healthy children living in Klintsy. This fact cannot be explained by influence of disease because there was no difference in cytogenetical markers between the groups of healthy children and children with thyreopathology from Moscow. The number of dicentrics was increased in cells from children with high level of internal contamination (more than 400 nCi) living in Krasnaya Gora (15 40 Ci/km2). It is proposed that the internal irradiation is more important for the CA induction. The symmetrical translocation analysis using the method of fluorescence in situ hybridisation (FISH) has shown the absence of this type of aberrations in cells from children with high level of internal irradiation whereas the translocations frequency in cells from control children was (1.1 +/- 0.4) per 1000 cells. PMID- 7489093 TI - [The frequency of chromosome aberrations in the lymphocytes of children with thyroid diseases living in Bryansk Province]. AB - The number of unstable chromosome aberrations was determined in peripheral blood lymphocytes from 15 children with thyreopathology living in Klintsy (Bryansk Province); the mean age was 14 years. The number of dicentric chromosome is determined to be (0.18 +/- 0.07) per 100 cells and is significantly higher than that in cells from healthy children of Klintsy and Moscow (control). There was no differences in the frequency of other types of chromosome aberrations as well as of aberrant cells. PMID- 7489090 TI - [The results of the cytogenetic examination of children and adolescents living in radionuclide-contaminated regions of Bryansk Province]. AB - The paper presents the data on cytogenetic studies of 663 children inhabited radionuclide contaminated territories of Bryansk Province. Statistically significant difference with control was observed only for the territory with the highest radioactive contamination level of 137Cs (1000 kBq/m2). For two other territories with lesser contamination (100 and 700 kBq/m2) the rate of chromosome aberrations is nearly the same and does not differ significantly from control one. No difference in the level of cytogenetic damage was discovered for children affected by radiation pre- or postnatally. PMID- 7489094 TI - [The cytogenetic examination of children in the Saint Petersburg region who suffered as a result of the accident at the Chernobyl Atomic Electric Power Station. The frequency of unstable chromosome aberrations in the peripheral blood lymphocytes]. AB - Cytogenetic study of 90 children suffered from the Chernobyl accident was carried out. The rate of unstable chromosome aberrations in lymphocytes was higher in groups of children whose parents took part in amelioration work after the Chernobyl accident, children evacuated from the contaminated areas and children of patients received antitumor radio- and chemotherapy as compared to nonexposed children. The increase was due mainly to single fragments. In the group of children evacuated from contaminated areas the rate of exchanges was also increased. PMID- 7489095 TI - [Stable chromosome aberrations in the lymphocytes of the peripheral blood in persons who suffered as a result of the accident at the Chernobyl Atomic Electric Power Station]. AB - The people suffered from Chernobyl accident exhibited the increased frequency of stable chromosome aberrations in peripheral lymphocytes as it was estimated by two-colour fluorescence in situ hybridisation with whole chromosome specific DNA probes for chromosomes 1, 4 and 12. However, the age dependence of this parameter in control group has to be estimated before the connection of this effect with radiation factor will be possible. PMID- 7489096 TI - [The genetic consequences of the Chernobyl accident. The monitoring of congenital developmental defects in newborn infants in Kaluga Province]. AB - The study concentrates on the genetic after-effects of the Chernobyl accident in some districts of Kaluga Province. The frequencies and range of congenital malformations, prematurely death-rate in the radionuclear polluted districts were compared with the control districts. Prematurely death-rate in compared regions is the same. The increase of the congenital malformation frequency is revealed in one the polluted districts (256/10,000) as compared with the control one (27/10,000). We assume that the observed effect is mainly connected with the professional activity of the population. PMID- 7489097 TI - [The cytogenetic effects in persons who suffered as a result of the accident at the Chernobyl Atomic Electric Power Station]. AB - Frequency of chromosome aberration was evaluated in 537 persons taken part in amelioration after the accident. The highest rate of aberration was found in covering builders and dosimetric: 3.24 +/- 0.25 and 3.11 +/- 0.43 per 100 cells, respectively. The mean rate of aberrations among the Chernobyl NPP staff was 2.37 +/- 0.20 per 100 cells, in the other examined groups the mean yield of aberration varied from 1.31 to 1.47 per 100 cells. The found aberration rates correspond to the equivalent whole body doses in the range from 131 to 515 mGy as evaluated by the established dose-response curve. In the group of covering builders the individual aberration rates varied more markedly, and corresponded to the equivalent whole body dose up to about 1 Gy. Slides of 27 individuals were checked by an automated dicentric scoring system. The results showed a satisfactory correlation between the frequencies of dicentrics per chromosome detected by routine and computer methods. PMID- 7489098 TI - [The use of the FISH method for reconstructing the absorbed doses received by participants in the clean-up of the accident at the Chernobyl Atomic Electric Power Station]. AB - The frequency of stable translocations using two-color fluorescence in situ hybridisation (FISH) were determined in peripheral blood lymphocytes of 20 persons who took part in liquidation of the Chernobyl accident and of 11 persons of control group. The individual doses for all patients were estimated from a calibration dose-response curve for stable translocation. The doses estimated by our cytogenetic data not always coincided with official dosimetric data. The possibility of using FISH for dose reconstruction of past radiation exposure was discussed. PMID- 7489099 TI - [The adaptive response in the lymphocytes of children from an area with an elevated background of ionizing radiation]. AB - The ability to form the adaptive response in lymphocytes of 23 children, exposed to small radiation doses as the result of Chernobyl accident was studied by hydroxylapatite chromatography of cell lysates. No correlation was found between the ability to form adaptive response and received radiation dose. PMID- 7489100 TI - [The radioadaptive response in the lymphocytes of schizophrenia patients]. AB - Radioadaptive response in cells from three patients was studied by induction of structural chromosome aberrations. Radioadaptive response was revealed in lymphocytes of all three patients. Coefficient of protection was 59.0% and did not differ from this after cell pretreatment with interferon. It is suggested the similarity or identity of mechanism of radioadaptive response and of interferon antimutagenic action which was implemented by non-repair way. PMID- 7489102 TI - [The genetic consequences of the radioactive contamination of populations of Arabidopsis thaliana growing in a 30-kilometer area of the accident at the Chernobyl Atomic Electric Power Station]. AB - Annually during six years since 1986 dynamics of the mutation load in Arabidopsis populations was studied. Frequency of the embryonic lethal mutations was studied with Muller embryonic test. First two years after the accident an increase of mutation load in Arabidopsis populations was observed at all levels of radioactive contamination. In the areas with contamination up to 10 mR/h the mutation load decreased to the control level by 1990. In the areas with contamination up to 130 mR/h the mutation load exceeds the control by 4-8 times. PMID- 7489101 TI - [The cytogenetic analysis of the adaptive response in the lymphocytes of donors living in areas with different levels of radioactive contamination]. AB - The adaptive response was studied in peripheral lymphocytes of healthy donors residing at territories with various levels of radioactive contamination. For donors from a clean territory (the town of Obninsk), the adaptive response depended on the cell cycle stage at which lymphocytes had been exposed to adaptive and challenge doses. The most expressed adaptive response was observed if lymphocytes had been exposed to the adaptive dose 0.05 Gy at G0 or G1 stage and to the challenge dose 0.5 Gy at G2 stage. Lymphocytes of donors from a contaminated territory (the town of Novozybkov) did not show an adaptive response in the conditions described above. PMID- 7489105 TI - [The genetic effect in Pinus sylvestris L. populations of the eastern Urals radioactive trace from the control area of the accident at the Chernobyl Atomic Electric Power Station and from the region of the nuclear device tests at the Semipalatinsk proving grounds]. AB - The method of isoenzyme electrophoresis was used for comparative evaluation of genetical effects in populations of Pinus sylvestris L. growing in Kyshtym region (east Urals), the Chernobyl accident zone and in the region of Semipalatinsk nuclear tests (Altai). It was shown that mutation frequencies for allozyme loci are ranged as follows: Ural > Altai > Chernobyl. PMID- 7489104 TI - [The cytogenetic effects in natural populations of Crepis tectorum exposed to chronic irradiation in the region of the Chernobyl Atomic Electric Power Station. An analysis of the frequency of chromosome aberrations and karyotype changes in the 3rd and 4th years after the accident]. AB - In three natural Crepis tectorum populations from 30-km zone of the Chernobyl NPP cytogenetic analysis of plant progenies was carried out. Germination capacity was not correlated with the dose rate of chronic irradiation. There was a correlation between frequency of cells with chromosome aberrations and frequency of seedlings with non-identical altered karyotype among which pericentric inversions and reciprocal translocations dominated. 9-chromosome altered karyotypes were found. Homozygous plants with altered karyotypes were not observed. Several heterozygous plants segregated homozygous seedlings with altered karyotype were found. In comparison with the second year after the accident in the third and fourth years there was stronger deviation from linear dependence between the cytogenetic effect and the dose rate of irradiation. PMID- 7489106 TI - [The genetic processes in chronically irradiated natural populations of Centaurea scabiosa L. growing in the eastern Urals radioactive trace]. AB - The examination of chronically irradiated natural populations of C. scabiosa L. have been carried out for 30 years at the territory of the east Urals radioactive trial. By using of different tests (frequency of aberrant cells, chlorophyll mutations and mutations in Lap locus) the mutation rates in chronically irradiated populations and in the control natural populations were compared. It was shown that long-term radiation influence caused changes in population genetic structure due to induction of new allelic forms and selection of more adapted genotypes. PMID- 7489103 TI - [The monitoring of natural populations of Dactylis glomerata L. in the area of the accident at the Chernobyl Atomic Electric Power Station]. AB - Results of five years monitoring (1986-1991) of seed quality (SQ) and chlorophyll mutation frequency (MF) of Dactylis glomerata L., growing within 30-km zone of Chernobyl NPP are presented. It was shown that during initial postaccident period (1986, 1987) the criteria depend of the gamma-phone on the sampling plots. It was not found the correlation between SQ, MF and duration of plant growing under condition chronic irradiation. PMID- 7489107 TI - [The cytogenetic effects in Crepis tectorum populations growing in Bryansk Province observed in the 7th year after the accident at the Chernobyl Atomic Electric Power Station]. AB - Cytogenetic analysis of seedlings from the seeds of four populations of Crepis tectorum plants grown during 7 years in localities of Bryansk Province contaminated with radioactive products was carried out. The frequency of chromosome aberrations in root meristematic cells was higher than at similar dose rates within 30-km zone of the Chernobyl NPP. At the same frequency of chromosome aberrations (induced at higher dose rates) karyotypes with pronounced frequency were observed in 30-km zone of the Chernobyl NPP. In Bryansk Province karyotypes were not found. This fact can be explained by the following: irradiation which induced the bulk of chromosome aberrations took place not at the stages of meiosis-early embryogenesis when the induction of chromosome aberrations leading to changed karyotype was more possible but after that. PMID- 7489109 TI - [The effect of inhibitors of DNA and protein synthesis on the yield of chromosome aberrations in a human lymphocyte culture under gamma and neutron irradiation at different stages of the mitotic cycle. The cytogenetic effects at the G1 stage]. AB - The impact of DNA of protein synthesis inhibitors (cycloheximide or hydroxyurea with 5-fluorodeoxyuridine -HU + FUrd, respectively) on the yield of chromosome aberration induced with gamma rays (1 or 2 Gy) and neutrons (0.5 or 1 Gy) at G1 stage of mitotic cycle was studied in human lymphocytes. The both inhibitors were discovered to increase the effect of gamma radiation at nearly all periods after irradiation during 5 hours. Conversely, the decrease in the level of chromosome aberrations was observed for neutrons after action of inhibitors at the same periods. PMID- 7489110 TI - [A cytogenetic analysis of the combined action of pesticides and irradiation on human lymphocytes]. AB - The efficiency of the combined action of pesticides and irradiation at the G(o) stage was studied in cultured human lymphocytes. Carbophos (malathion) increased the yield of chromosome and chromatid fragments in irradiated lymphocytes. Herbicide 2,4-D (dichlorophenoxyacetic acid) raised lymphocyte radiosensitivity by increasing the yield of chromosome type aberrations; the radiosensitizing effect of the herbicide decreased as its concentration increased. PMID- 7489108 TI - [The anticlastogenic effect of ascorbic acid in relation to the damages induced in human lymphocytes by the photomutagenic action of 8-methoxypsoralen and by ammonium molybdate]. AB - In experiments with cultured human lymphocytes anticlastogenic effect of ascorbic acid (AA) with respect to photomutagenic action of 8-methoxypsoralen (8-MOP) and ammonium molibdenate (Mo) was studied. It was found that AA effectively protected from mutagenic action of Mo. Effective protection of cells from 8-MOP monoadducts and DNA interstrand links was also observed. PMID- 7489111 TI - [UV-induced unscheduled DNA synthesis in the peripheral blood lymphocytes of participants in the clean-up of the aftereffects of Chernobyl accident]. AB - The dependence of UV-induced unscheduled DNA synthesis on UV exposure dose was studied in intact donors and persons dealt with liquidation of the Chernobyl accident consequences in 1986-1990. In the group of persons exposed to high doses the decrease in the level of UV-induced unscheduled DNA synthesis and the increase in the level of spontaneous one was registered. PMID- 7489112 TI - [The antimutagenic activity of ternary diallyl copolymers]. AB - Antimutagenic activity of triple copolymers of diallyl origin was investigated by animal cell test (mouse bone marrow erythrocytes, 1.5 Gy of gamma irradiation) and by plant cell test (seeds of barley, 5 Gy of gamma irradiation). Effective protection of genetic structure was achieved owing to combination of moderate antimutagenic activity of the polymer matrix and scavenging ability of sterically hindered phenols in the polymer side chain. PMID- 7489113 TI - [The results of multiyear research on fish populations from reservoirs falling within the area of the Kyshtym accident (a review)]. AB - The results of long-term studies of fish populations of various kinds from lakes polluted with radionuclides as a result of the Kyshtym accident are considered. The literary and own data on embryonic development studies with cytogenetic analysis, results of morphometric and morphological studies of fish populations, investigations of population structures by electrophoretic analysis of markers of structural genes were analysed. Critical comparison of the data obtained by the various authors in different years was conducted. The conclusion was made that none of the tests showed unequivocal effect of radionuclide contamination on ichthyofauna. PMID- 7489114 TI - [The radioprotective action of efazol under different conditions of irradiation of the germ cells of male mice]. AB - A study was made of the protective effect of ephasol against genetic damages induced in mice germ cells by single, fractionated and chronic gamma radiation. Study of male reproductive ability, frequency of induced dominant lethal mutations, abnormal sperm heads and reciprocal translocation show that the efficacy of ephasol was higher in condition of chronic exposure. PMID- 7489116 TI - [The consequences for the first-generation progeny of Wistar rats of the irradiation of both parents with a background of mexamine use]. AB - In experiments with Wistar rats it was shown that the efficiency of mexamine as radioprotector was substantially lower when sex cells of both parents (spermatozoids, spermatids, ovocytes) were irradiated with a dose of 2-4 Gy than after irradiation one parent only. It may be associated with the aggravation of effects in the posterity being conceived from gametes of both exposed parents. PMID- 7489117 TI - [The effect of dosage strength on the induction of chromosome aberrations and lung tumors in mice under the combined action of ionizing radiation and chemical substances]. AB - In the experiment with combined influence of single or prolonged gamma irradiation (35 cGy) and chemical agents (urethanum, nitrogen oxide) on the induction of chromosome aberrations in bone marrow cells and lung tumor the effect of dose rate was studied. The frequency of chromosome aberrations increased with dose rate. Yet the correlation between the ratio of irreparable (stable) aberrations and dose rate was of inverse direction. The character of variations in lung adenoma frequency under multifactor influence in the case of prolonged irradiation (increase with dose rate decrease) supports the correlation between the level of stable aberrations in the pool of genetic lesions and the frequency of lung adenoma. PMID- 7489115 TI - [Modification of the genetic effect of radiation in mice exposed in the region of the Chernobyl Atomic Electric Power Station]. AB - A study was made of the protective effect of beta-carotine and ephasol against genetic damages in germ cells of male mice, exposed within the zone of Chernobyl NPP or ingested by 137Cs. The analysis of reproductive function, frequencies of dominant lethal mutations, abnormal sperm heads, reciprocal translocations and testis mass decrease show that both agents are more effective in protection against reproductive function damages than against genetic effects. PMID- 7489118 TI - [The postnatal development of the progeny of males whose germ cells were irradiated at different stages of spermatogenesis]. AB - Distinct genetic radiosensitivity of germinal cells of males irradiated during different stages of spermatogenesis with doses of 0.25-5.0 Gy leads to reduction in vital newborn rats number in the first generation progeny and to elevated postnatal mortality rate. These postnatal ontogeny disorders depend on the irradiation dose and spermatogenesis stage for a moment irradiation. PMID- 7489119 TI - [The use of natural sorbents for removing radioactive cesium from the body of experimental animals]. PMID- 7489121 TI - Perspectives on the role of afferent signals in control of motor neuroprostheses. AB - K.O. Johnson reviews the architecture and low level neural mechanisms by which the external environment is transduced and encoded into the neural system, summarizing work that correlates neurophysiological and psychophysical testing with isolation of sensory components. The slowly adapting Type I afferent system is responsible for form and texture perception; the rapidly adapting afferent system is responsible for motion perception; and the Pacinian corpuscle system is responsible for vibratory sensation. R.R. Riso reviews the current level of understanding of the major factors to be considered in the design of a functional neuromuscular stimulation (FNS) grasp controller that uses cutaneous sensory feedback to detect slip. The elegant natural control scheme that matches the ratio of grip and lift forces to frictional conditions provides a model for implementing a slip-based control algorithm. D. Popovic discusses the possible use of recordings from more proximal peripheral nerves to determine needed information for synthesis of locomotion. The discussion is illustrated with an animal model where rule-based closed-loop control is used for the ankle joint during treadmill locomotion. Neural signals from the tibial and superficial peroneal nerves were employed to substitute for missing afferent input from cutaneous and proprioceptive sensors. The feasibility of more invasive intraneural electrodes for distinguishing sensory from motor information in mixed nerves is considered. M. Koris raises surgical and functional issues relevant to developing clinical FNS systems. C. Van Doren suggests alternative neurophysiological and engineering approaches. PMID- 7489120 TI - Glucocorticoid resistance in asthma. PMID- 7489122 TI - A review of the in vitro evaluation of conduit-mounted cardiac valve prostheses. AB - This review concerns the issues affecting the in vitro evaluation of conduit mounted prosthetic heart valves at the design development stage, and the question of standardisation of testing at the quality assurance stage. Particular attention is given to areas of conduit valve development and research which have been neglected, ambiguously covered or left to the discretion of the researcher in the current standard for conventional prosthetic valves, ISO 5840. PMID- 7489123 TI - Walking when utilizing a sensory feedback system and an electrical muscle stimulation gait orthosis. AB - Electrical muscle stimulators (EMS) have been combined with a reciprocating gait orthosis (RGO) to produce an EMS-RGO system for constant velocity ambulation exercise in a spinal cord injured (SCI) individual. The objective of this study is to evaluate an auditory feedback system (AFS) used by the SCI subject when ambulating with the EMS-RGO. Three different types of auditory signals (M = metronome alone, MSL = metronome plus stride length information, and NONE = no auditory feedback) were examined at three different constant walking velocities (WV1 = 0.64 kph, WV2 = 1.22 kph, and WV3 = 1.80 kph). The experimental design was 3 x 3 full factorial with repeated measures, and the dependent variable was the absolute error (AE) of walking distance (absolute value of the distance the subject walked in one minute subtracted from the target distance). A block diagram and circuit schematic of the AFS is provided. The results indicate that at WV1 there is no significant difference of AE among the three auditory conditions (M, MSL and NONE). At WV2, the AE for the NONE auditory condition is significantly greater than the AE for M and MSL (p < 0.05). At WV3, the AE for the NONE auditory condition is significantly less than the AE for M and MSL (p < 0.05). Finally, there was no significant difference in AE between both the M and MSL auditory conditions at any of the three walking velocities. This study indicates that an adequate sensory feedback system for constant velocity control of ambulation in this SCI subject consists of metronome only auditory feedback.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7489124 TI - Separation of fetal and maternal ECG complexes from a mixed signal using an algorithm based on linear regression. AB - Fetal scalp ECG and Doppler ultrasound are the two methods universally used to derive the fetal heart rate (FHR) for cardiotocography. Other fetal signal sources have not been used successfully because of the complexity of the signal and the need for an additional maternal ECG input. In this paper, we present an algorithm for separating the fetal and maternal ECG signals obtained from intrauterine electrodes during labour. The algorithm detects all occurrences of ECG complexes and uses linear regression functions to compare each complex with a set templates. Sets of templates are identified as either maternal or fetal in origin and two signals are output for heart rate measurement. The outputs are also processed to eliminate artefacts that may occur when the maternal and fetal complexes are coincident. The algorithm processes 10 seconds of data at a time (in about 200 ms on a standard PC) while a further 10 s of data is being acquired. It has the advantage that no extra input is required, such as maternal ECG obtained from chest leads, and only assumes that two different populations of complexes of two different shapes are present in the input signal. The algorithm could also be used for the separation of fetal and maternal signals obtained from abdominal electrodes. PMID- 7489125 TI - Assessment of normal testis growth by quantitative texture analysis of 2-D echo images. AB - Six phases summarize the evolution of the healthy male gonad. Qualitative examination of two-dimensional echograms detects tonal differences at different ages of the testes. However, routine qualitative evaluation of the echograms is too subjective and dependent upon the scanner controls; on the other hand, simple morphology reports are not sufficient to establish the correct histologic and physiologic evolution. So far computer-assisted quantitative analysis of ultrasonic images has been successful in the characterization of tissues based on their echographic properties; thus this approach could be suitable for an accurate identification of the age of healthy testis. Sixty-two normal subjects, ranked in six groups according to age, underwent echographic examination in standard conditions. Echograms were digitized and analysed by means of well established algorithms for first and second order statistics. Significant differences in tonal features were found able to discriminate among different ages, in agreement with the qualitative analysis and the histologic reports. Furthermore, some second order analysis features exhibit statistically significant alterations of the micro-structural organisation of the echo patterns consequent to changes of the configuration of the ultrasonic targets, that is, of the gonad's histology. In conclusion, quantitative analysis of echograms can provide a tool for a better assessment of normal testis evolution than the qualitative approach. PMID- 7489126 TI - Evidence that maximum muscle stress is not a constant: differences in specific tension in elbow flexors and extensors. AB - The specific tension of muscle (or maximum muscle stress) is the maximum force developed per unit cross-sectional area and is a frequently used parameter by investigators estimating muscle force. Generally, it is assumed to be a constant value for all muscles and, when multiplied by a muscle's cross-sectional area, is used to provide a measure of a muscle's maximum force production. In this study, the specific tension for elbow flexors and for extensors were compared to evaluate the validity of this assumption. Maximum muscle stress was determined using maximum joint moments measured as a function of joint angle and using anatomical parameters reported in the literature. It was observed that the specific tension for elbow flexors was considerably larger than for extensors when measured a variety of ways. The exact reasons for the differences are unknown, but variations in specific tension of individual fibers may play a role. It was concluded that the use of a constant value for specific tension in muscle models is questionable in studies that demand accurate results. PMID- 7489127 TI - A method for the evaluation of tensile properties of skin equivalents. AB - In vitro production of anchored skin equivalent is a new therapeutical option for burn patients. A skin equivalent is a combined culture of dermal and epidermal layers. The dermal layer provides important mechanical properties, such as tensile resistance and nonlinear elasticity, to the skin equivalent during its development. Prior to any in vivo human transplantation, the tensile properties of cutaneous equivalents have to be evaluated as a function of its structural components, in view of establishing the culture conditions leading to the best mechanical resistance and stretchability characteristics. However, the handling and clamping of skin equivalents are frequent causes of tearing and lack of repeatability in the measuring of tensile properties. A new indentation method involving a specially designed culture dish has been developed to minimize the risk of damage. Using this new culture dish, cutaneous equivalents were installed on an indentation apparatus. The central loading of a spherical tip was transmitted to the central area of a circular anchored cutaneous equivalent and was recorded with tip position. The tests were achieved at a constant low deflection rate of the tip. This new and accurate method gave repeatability in three central load-deflection characteristics of anchored dermal equivalent: the high-modulus (0.15 g mm-1), the central load of rupture (1.49 g), the rupture deflection (0.470 mm). This indentation test is expected to be an efficient tool in the evaluation of various skin equivalent models tensile properties. PMID- 7489128 TI - Cancellous bone stresses surrounding the femoral component of a hip prosthesis: an elastic-plastic finite element analysis. AB - The cancellous bone stresses surrounding the femoral component of total hip replacement were investigated using the finite element method. Four versions of a certain femoral hip prosthesis (the Freeman, Corin Medical), cemented, HA coated, press-fit, and press-fit with ridges were analysed. Each model was subjected to two separate load cases with the cancellous bone modelled as an elastic perfectly plastic material. The effect of bone quality was investigated by varying the cancellous bone stiffness. The resulting cancellous bone stress distributions were compared to that of the intact femur. The results were also compared to clinical subsidence data (published elsewhere) for the Freeman femoral prosthesis to determine if the initial cancellous bone stress distribution could be used to predict the migration of the various versions of this prosthesis. The results showed that the press-fit designs of prosthesis generated substantially higher cancellous bone stresses than the cemented and HA coated designs, and that these stresses were up to 6.5 times higher than found in the intact femur. For all forms of fixation the cancellous bone stress distribution was found to be insensitive to changes in the trabecular bone stiffness: thus poor quality cancellous bone is more likely to promote 'plastic' deformation, and therefore subsidence of the prosthesis. Comparison with the clinical migration data showed a good correlation and revealed that it may be possible to use the calculated initial cancellous bone stresses to predict the migration of the implant, and hence the probability of early and mid-term aseptic loosening.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7489129 TI - A system for rapid identification of respiratory abnormalities using a neural network. AB - Potential victims of Sudden Infant Death Syndrome (SIDS) can usefully be monitored in the home environment. Conventional respiration movement monitors can be helpful but may not detect potentially dangerous hypoxaemic episodes. Thus oxygen monitoring is to be preferred but can be difficult to use in the home. In an attempt to overcome these difficulties this paper presents the results of an exploratory experiment into the use of a neural network to link the output of a respiration pressure monitor to the classification of breathing patterns as effective or otherwise. It has been shown that it is possible to predict changes in oxygen saturation, which could signify potentially dangerous episodes earlier than when other methods are employed. PMID- 7489130 TI - [Selection of criteria for evaluation of septic conditions]. AB - In a prevalence study of septicaemia at a surgical clinic 645 patients were followed up. The assembled data were compared with data from a similar study made in 862 patients which was implemented in different departments of Ostrava hospitals. In the two studies different criteria of sepsis were used. In the study conducted at the surgical clinic these criteria were more precisely specified and extended. This trebled the assessed number of sepses. From comparison of results of these studies ensues that more frequent bacteriological monitoring of patients in necessary. PMID- 7489131 TI - [Diagnosis of acquired toxoplasmosis using simultaneous determination of specific immunoglobulins M, A and E]. AB - Simultaneous estimation of specific immunoglobulins M,A,E was tested in sera of 57 patients with confirmed acquired toxoplasmosis (24 acute and 33 subacute or chronic) using immunosorbent agglutination assay (ISAGA) and immunofluorescent tests (toxoplasma antibody-total and IgM). The antigen for ISAGA was prepared from tissue cultures HeLa infected with Toxoplasma gondii virulent strain P. Toxoplasmas were maintained in tissue cultures continuously for 18 years. High levels of IgM antibodies were found in all 24 patients during the acute phase of toxoplasmosis (0-5 month after onset). IgA antibodies were found also in all but one and IgE in 71% of these patients. IgM antibodies persisted for the longest period and it was possible to find them in 14% of patients 3 years or longer after the onset, IgA antibodies persisted for 9 months similarly as IgE (with one exception after 2 years). Simultaneous estimation of specific immunoglobulins M,A,E combined with immunofluorescent tests makes the diagnosis of Toxoplasma infection more reliable. PMID- 7489132 TI - [Occurrence of Klebsiella pneumonia strains resistant to third-generation cephalosporins at the Pediatric Clinic of the Medical School Hospital in Olomouc]. AB - The authors describe the high incidence of strains of Klebsiella pneumoniae resistant to cephalosporins of the IIIrd generation at the department of pathological neonates of the Paediatric Clinic of the Faculty Hospital in Olomouc. They discuss possible reasons for this adverse situation and the way how to resolve it. They emphasize the necessity of rational administration of antibiotics and continuous monitoring of resistance in the most frequent causal agents of bacterial infections. PMID- 7489134 TI - [Occurrence of Salmonellae in the laughing gull (Larus ridibundus)]. AB - In 1984-1994 black-headed gulls (Larus ridibundus) from various localities in the Czech Republic were examined to assess the incidence of salmonellae. A total of 99 eggs from two nest colonies were examined. No salmonellae were detected. In examinations of 740 young birds from three colonies the total prevalence was 19.3%. In full grown gulls (number of examined birds 189) from five localities the total prevalence of salmonellae was 4.2%. In water specimens near the nest colonies, other habitats of the gulls and from soil specimens from one nest colony (throughout the year) salmonellae of the same serotypes as in gulls were isolated. The young birds contaminate surface waters with salmonellae. The importance of adult birds as a source of infection is much smaller. Only for the young of aquatic birds (fowl or free living) gulls could be an important source of salmonella infections. The presence of salmonellae in gulls is due to contamination of surface waters and the surroundings of nest colonies with salmonellae. PMID- 7489136 TI - [Detection of HIV virus antibodies in saliva]. AB - The authors evaluate the use of saliva as alternative biological material for detection of HIV antibodies. If collected properly and when using the appropriate ELISA methods, HIV antibodies can be assessed in saliva with considerable sensitivity and specificity. Testing in saliva eliminates many disadvantages found when assessing HIV antibodies in serum. It does not require trained staff for blood sampling, collection and processing of saliva reduces the risk of professional infection to a minimum and can be carried out also under field conditions. A great advantage is the easy and unpretentious transport of collected samples to the laboratory, even long distance transport at extreme temperatures (transport medium in the testing kit Omni-SAL ensures great stability of the sample as well as proper collection of the sample). Alternative testing in saliva is useful in particular for epidemiological surveillance and for screening of HIV antibodies in population groups with a high risk of infections such as drug users and commercial works where it is often very difficult to obtain blood samples for examination. PMID- 7489135 TI - [Animal foci and transmission routes of Toxoplasma gondii]. AB - In the transmission of T. gondii from animal sources to man a major part is played by animal tissues containing toxoplasmatic cysts and toxoplasmatic oocysts which result from the sexual reproduction of the parasites in the small intestine of feline animals. Tachyzoits play a minor role in the development of toxoplasmosis in man, most frequently when animals, their tissues and secretions are handled in the stage of parasitaemia. As to domestic animals, rabbits and sheep are infected most frequently and to a very small extent cattle. Animals from small farms are infected more frequently as there is a greater chance of contact with the domestic cat. A natural reservoir of T. gondii are small terrestrial mammals where intrauterine transmission occurs and the infection passes from one generation to the next. Thus elimination of the parasites in animal sources is not feasible and in prevention of toxoplasmosis attention must be focused primarily on transmission of T. gondii from known animal sources to man. PMID- 7489133 TI - [The effect of subinhibitory concentrations of norfloxacin, erythromycin and roxithromycin on Pseudomonas aeruginosa virulence factors]. AB - The effect of sub-MICs (1/4 MIC, 1/8 MIC, 1/16 MIC) of norfloxacin, erythromycin, roxithromycin on Pseudomonas aeruginosa virulence factors was studied. It was found that 1/4 of the MIC of norfloxacin inhibited most effectively elastase as well as proteinase activity (to 30.1% or 29.2% of the control values). Macrolide antibiotics did not reduce the tested enzymic activities but caused their increase. PMID- 7489137 TI - [Staphylococcus aureus and the human immune system]. AB - The author characterizes factors of virulence of Staphylococcus aureus and its different interactions with immune systems of the host. She describes the immune mechanisms which play a key role in the elimination of staphylococcal infection and where inadequate function leads to the development of staphylococcal disease. The author draws attention to the complicated interrelationship of the complex of different virulence factors of staphylococcal strains involved in inhibition and dysregulation or in stimulation of effector mechanisms of immunity. PMID- 7489138 TI - [70 years' of the National Health Institute in Prague]. PMID- 7489139 TI - The corporate influence. PMID- 7489140 TI - Introducing graft-versus-leukemia into autologous stem cell transplantation. AB - Although autologous transplantation of stem cells is widely used to support high dose chemotherapy and radiation for the treatment of hematologic malignancies, relapse is a severe impediment to its wide use, especially in patients receiving transplants for leukemia. This is related to the infusion of leukemic cells in the autograft and the lack of an allogenic graft-versus-leukemia (GVL) effect. This review discusses the current approaches that are under study to introduce a GVL effect into autologous transplantation. PMID- 7489141 TI - The role of natural killer cells in the treatment of chronic myeloid leukemia. AB - The success of chemotherapy in patients with leukemia whose marrow appears to be replaced by leukemia cells must be due to the persistence of normal stem cells. In this normal population are the progenitors of the cells of the immune system. Natural killer (NK) cells originate in the bone marrow. On maturation and activation with interleukin 2 (IL-2) or other cytokines, NK cells develop cytotoxic activity against a variety of leukemic blasts, including those from patients with chronic myeloid leukemia (CML). In the past few years, bone marrow transplantation (BMT) and alpha-interferon (IFN-alpha) have proved to be the most promising therapies for the treatment of CML. In both these therapies, NK cells may play a prominent role. In this article, we discuss the antitumor/antileukemia activity of human NK cells, the presence of benign NK cell precursors in the different stages of CML, the role of NK cells in BMT and IFN-alpha treatment, and the potential therapeutic applications of NK cells in patients with hematologic malignancies. PMID- 7489142 TI - Collagen matrix: an attractive alternative to agar and methylcellulose for the culture of hematopoietic progenitors in autologous transplantation products. AB - Autografts using untreated or in vitro manipulated bone marrow and peripheral blood stem cells represent promising approaches to the treatment of malignant diseases. In this work, the collagen gel culture technique was compared with agar and methylcellulose for its capacity to permit the growth of human granulomonocytic (day 14 CFU-GM; collagen vs agar or MTC) or erythroblastic (day 7 CFU-E and day 14 BFU-E; collagen versus methylcellulose) colonies in autologous transplantation products. Our results show that the collagen culture system always gave as many or more colonies than the other techniques. It also allowed harvesting of gels onto glass slides and subsequent May-Grunwald-Giemsa, cytochemical or immunocytochemical staining. We suggest that the collagen assay represents an interesting alternative to the widely used agar or methylcellulose systems for the culture of hematopoietic progenitors because of the equal or higher number of colonies detected, the easy phenotypical identification of colonies in stained gels, and the ability to store high-quality documentation. This technique is particularly attractive for use in the quality control of autologous bone marrow transplantation procedures. PMID- 7489143 TI - Maturation of mobilized peripheral blood progenitor cells: preclinical and phase I clinical studies. AB - The use of mobilized peripheral blood progenitor cells (PBPC) after high-dose chemotherapy has markedly decreased the period of severe neutropenia. In an attempt to further decrease the duration of neutropenia, the potential of PBPC to mature during in vitro culture was assessed, with special attention being paid to culture medium, growth factors, and cell concentration. Concentrations of 10(6) PBPC/mL resulted in better recovery than 10(7)/mL as far as total cells, CFU-GM, and granulocytes were concerned. The combination of IL-3 + GM-CSF+G-CSF appeared to be better than any of these growth factors alone. Simple media, such as Medium 199, gave poorer cell recovery than more complex media, such as IMDM. With 10(6)/mL nonenriched PBPC in IMDM with IL-3/GM-CSF/G-CSF, on day 15 CFU-GM reached 450% of the initial level. At that point, granulocytes had increased 15 fold. A small phase I study was performed to assess the toxicity of infusing 1000 2000 mL of PBPC cultured for 3 days at 3-10 x 10(6)/mL with IL-3/GM-CSF/G-CSF in LifeCell bags. Although no clear decrease in the duration of neutropenia was observed, the infusions were uncomplicated in 5 of the 6 patients and had minor side effects in the sixth patient. These data suggest that in vitro differentiation of nonenriched PBPC is possible. However, to develop a clinically applicable method, several logistical problems will have to be overcome. PMID- 7489144 TI - Principles of bone marrow processing and progenitor cell/mononuclear cell concentrate collection in a continuous flow blood cell separation system. AB - The application of continuous flow apheresis technology to processing bone marrow for collection of the mononuclear progenitor cell population appears to follow the same principles as collection of mononuclear cells from peripheral blood. Unlike peripheral blood, however, where mobilization of cells from extravascular sites during the procedures contributes significantly to the final cell yield, the entire quantity of progenitor cells available for recovery from marrow is present in the original marrow when it is pooled. The process then becomes one of attempting optimal recovery of the cells of interest while excluding contaminating erythrocytes and cells of the myeloid series. This study reports the development of a protocol for recovery of MNC, CD33+, CD34+, and CD34+/DR- cells from harvested marrow for autologous and allogeneic transplants using a continuous flow blood cell separator, the variables influencing the recovery of the cells of interest and the clinical response to infusion of the processed cells. PMID- 7489145 TI - Autologous transplantation with peripheral blood stem cells in children and young adults after myeloablative treatment: nonrandomized comparison between GM-CSF and G-CSF for mobilization. AB - Recombinant human granulocyte-monocyte colony-stimulating factor (rhGM-CSF) was compared with recombinant granulocyte colony-stimulating factor (rhG-CSF) in 34 patients for mobilization of peripheral blood stem cells (PBSC) and for posttransplantation use. Peripheral blood stem cell mobilization was initiated by a single 1 h infusion of cyclophosphamide (4 g/m2) in all patients, followed by either a continuous infusion of rhGM-CSF (250 micrograms/m2/day) in 17 patients (group A) or a daily subcutaneous injection of rhG-CSF (10 micrograms/kg/day) in 17 patients (group B). PBSC were collected using a Fenwal CS 3000 continuous flow blood cell separator in one to three sessions. All patients suffered from various childhood malignancies. No difference in the number of collected cells among both groups was found. A mean of 2.7 x 10(8)/kg mononuclear cells (MNC) and of 7.9 x 10(4)/kg CFU-GM (colony-forming unit-granulocyte-macrophage) were collected in group A. In group B, 2.3 x 10(8)/kg MNC and 11.8 x 10(4)/kg CFU-GM were collected. In 33 patients, PBSC were reinfused after myeloablative therapy. Patients of group A (n = 17) were treated with rhGM-CSF (250 micrograms/m2/day) starting day +1, and patients in group B (n = 16) were treated with rhG-CSF (10 micrograms/kg/day) i.v. All patients showed a rapid and complete hematopoietic recovery without significant differences in both groups. Time to achieve 0.5 x 10(9)/L granulocytes was 10.9 days in group A and 11 days in group B.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7489146 TI - Use of the CS-3000 Plus to prepare apheresed blood cells for immunomagnetic positive cell selection. AB - The Baxter CS-3000 Plus Blood Cell Separator was used to prepare progenitor cell components in a three-step process. The process was designed to remove platelets and plasma from a leukapheresis cell product before incubation with anti-CD34 monoclonal antibody (9C5) and to remove unbound monoclonal antibody after incubation. Fluorochrome-labeled cultured KG1a (CD34+) cells were added to the leukapheresis cell product (at 2% of total WBC) before CS-3000 processing to evaluate the CS-3000 process for preparing cells for immunomagnetic selection using the Isolex 300 (SA) Magnetic Separation system. Data were obtained using five leukapheresis products. Two percent of the nucleated cells were lost during the platelet reduction wash, and 67% of the platelets were removed. The amount of antibody initially added to the cells for incubation ranged from 3.9 to 7 mg (0.5 microgram/10(6) nucleated cells). The residual antibody in the cells after the antibody wash ranged from 11 to 40 micrograms. The antibody wash resulted in a 3% loss of nucleated cells and an additional 66% removal of platelets. The antibody sensitized cells were then processed on the Isolex 300 (SA) system. The purity and yield of the Isolex KG1a cell product were 94% and 82%, respectively. PMID- 7489147 TI - The role of bone marrow transplantation in the treatment of myelodysplastic syndromes. PMID- 7489148 TI - Total mesorectal excision is optimal surgery for rectal cancer: a Scandinavian consensus. PMID- 7489149 TI - Mammary duct ectasia--periductal mastitis complex. PMID- 7489150 TI - Non-surgical trauma to the extrahepatic biliary tract. AB - Non-surgical trauma to the extrahepatic biliary tract is uncommon and encountered only rarely by surgeons outside specialist hepatobiliary centres. Such injury often results in significant morbidity and mortality. This review outlines the incidence, classification, mechanisms of injury, presentation, diagnostic techniques and management options. Depending on the type of biliary injury identified, an optimal method of repair is suggested. Recommendations are made for surgeons who encounter this type of injury and do not have the appropriate expertise for definitive management. PMID- 7489151 TI - Polyposis: the Peutz-Jeghers syndrome. AB - The classical pigmentation of Peutz-Jeghers syndrome distinguishes it immediately from the other polyposis syndromes. Less widely appreciated than this most obvious manifestation are the pitfalls in management presented by the risk of cancer and by the performance of multiple laparotomies that also characterize this condition. An outline of these risks and an approach that minimizes them is presented. PMID- 7489152 TI - Continent colonic conduit for the treatment of faecal incontinence associated with disordered evacuation. AB - A transverse colonic conduit incorporating an intussusception valve and skin flapped cutaneous aperture was constructed in nine patients with combined faecal incontinence and disordered evacuation. Intestinal continuity was restored with a colocolonic anastomosis. Median follow-up was 4 (range 2-15) months and daily irrigation with a median of 1.2 (range 0.3-2.0) litres of water resulted in evacuation in less than 1 h. At 1 month after operation there was no leakage of solid or liquid faeces from the anus between irrigations. The valve was continent to faeces and irrigation fluid, and no stoma appliances were required. PMID- 7489153 TI - Electrically stimulated colonic reservoir for total anorectal reconstruction. AB - Total anorectal reconstruction after abdominoperineal excision of the rectum has failed to achieve perfect continence. Electrically stimulated reservoir evacuation in combination with an electrically stimulated gracilis neoanal sphincter might improve results. A J pouch was constructed in an isolated colonic loop of seven dogs. Bipolar square wave pulses were delivered via two intramural stainless steel electrode pairs at 10 Hz. Stimulation parameters were varied to achieve adequate contraction. Serosal strain gauges recorded spontaneous and stimulated pouch motility. Evacuation was quantified by a volume displacement technique and observed fluoroscopically. Recordings were performed for a median of 3 (range 1-11) months. At 10 Hz and 0.5 ms pulse width, stimulation was required for 2 min and at voltages of 15 V (n = 4), 18 V (n = 1) and 20 V (n = 2) to obtain a contraction of amplitude comparable to that of a spontaneous contraction. Suprathreshold stimulation invariably resulted in colonic pouch contraction. The mean(95 per cent confidence interval (c.i.)) stimulus-response latency was 25.5(1.9) s. The mean(95 per cent c.i.) intraluminal pressure generated during stimulation was 114.1(17.0) cmH2O and 64.6(12.0) cmH2O during spontaneous activity (P < 0.001). In conclusion, electrical stimulation via intramural electrodes produced contraction generating sufficient intraluminal pressure to effect evacuation of a canine colonic pouch. This has potential for incorporation with an electrically stimulated neoanal sphincter in total anorectal reconstruction to improve evacuation and continence. PMID- 7489154 TI - Factors that influence functional outcome after coloanal anastomosis for carcinoma of the rectum. AB - Thirty consecutive patients had laboratory assessment of anorectal function after rectal excision and stapled coloanal anastomosis for rectal carcinoma. Eleven patients experienced perfect continence but 19 had faecal leakage with or without urgency of defaecation. Median (interquartile range) function was related to the pressures generated in the anal sphincter at rest (good versus poor function: 80 (63-91) versus 51 (23-60) cmH2O, P < 0.01), during maximum squeeze (160 (126-203) versus 102 (58-112) cmH2O, P < 0.01) and during reflex inhibition (58 (23-63) versus 36 (18-54) cmH2O, P < 0.05). Poor function was significantly commoner in women than in men (P < 0.01). These findings suggest that occult damage may have occurred to the anal sphincter before low anterior resection. Careful preoperative evaluation with manometry and endoanal ultrasonography may detect such damage and allow selection of patients for colopouch reconstruction. PMID- 7489155 TI - Defunctioning colostomy for low anterior resection: a selective approach. AB - Seventy-seven consecutive low anterior resections of the rectum were performed with a selective approach to the use of a defunctioning colostomy. A defunctioning colostomy was performed in seven patients (9 per cent) where there was concern about the anastomosis due to difficult dissection (three), incomplete doughnuts (three) and tension on the anastomosis (one). The mean level of the tumour in the defunctioned group was 7.6 cm. Clinical anastomotic leakage occurred in two patients (3 per cent) in the non-defunctioned group, both of which were controlled with subsequent transverse colostomies. There were no perioperative deaths. Selective defunctioning of low colorectal anastomoses can produce low rates of anastomotic dehiscence while reducing the morbidity associated with a temporary stoma. PMID- 7489156 TI - Detection of circulating tumour cells with the magnetic activated cell sorter. AB - A technique has been developed for the detection of tumour cells in blood with the magnetic activated cell sorter (MACS). Colonic carcinoma cell lines and disaggregated primary tumours were used to establish optimal conditions of separation. A murine monoclonal antibody specific for epithelial cells was added to the suspension of leucocytes and tumour cells, followed by magnetic labelled goat antimouse antibody. The labelled tumour cells were retrieved by passing this suspension through a MACS separation column in a strong magnetic field. Tumour cells were detected at a dilution of 10 cells per ml blood. Tumour cells were identified in mesenteric blood in three of 24 patients undergoing surgery for colorectal cancer. This study supports the use of the MACS to detect circulating tumour cells. PMID- 7489157 TI - Clinical impact of colonoscopic screening in first-degree relatives of patients with hereditary non-polyposis colorectal cancer. AB - Sixty-one asymptomatic individuals with an affected first-degree relative from five large hereditary non-polyposis colorectal cancer (HNPCC) kindreds were screened by colonoscopy. Neoplasms were found in nine (15 per cent) of 61 individuals on the first screen. Five subjects had a single adenoma while two had two adenomas each. There were two patients (3 per cent) with malignant neoplasms: one with a Dukes B adenocarcinoma and one with synchronous Dukes C adenocarcinomas in the caecum and ascending colon. These findings support the hypothesis that adenomas do not occur in large numbers in HNPCC families but, because of the high malignant conversion rate, biennial colonoscopy with removal of polyps seen is recommended. PMID- 7489158 TI - Lateral subcutaneous sphincterotomy for treatment of anal fissure in children. AB - Twenty-three children, aged between 8 and 168 months, underwent lateral subcutaneous sphincterotomy. All fissures healed by 8 weeks after operation. Two children had recurrent proctalgia and defaecating difficulties after surgery despite a healed fissure. Requirement for stool softener was reduced or abolished in 17 children. The parent-child satisfaction score after operation was more than 70 per cent in 19 of 23 cases. Lateral subcutaneous sphincterotomy is an effective procedure in children. PMID- 7489159 TI - Single-centre prospective study of isolated limb perfusion with melphalan in the treatment of subungual malignant melanoma. AB - Subungual melanoma is rare and experience in treating this condition with isolated limb perfusion is limited. Between 1985 and 1990, 24 patients were treated by digital amputation and isolated limb perfusion with melphalan and mild hyperthermia. The disease was staged according to the M.D. Anderson classification: stage I (17 patients), stage IIIA (one), stage IIIB (two) and stage IIIAB (four). Thirteen lesions were on the foot and 11 on the hand. Seven patients have developed locoregional recurrence. The estimated overall 2- and 5 year probabilities of survival were 77 and 46 per cent respectively, while for disease-free survival the rates were 58 and 51 per cent. When these results were compared retrospectively with those in 111 patients treated by amputation alone, no significant difference in survival was demonstrated. This experience suggests that isolated limb perfusion with melphalan and mild hyperthermia confers no additional survival benefit over appropriate surgery. PMID- 7489160 TI - Carbon dioxide laser ablation as an alternative treatment for cutaneous metastases from malignant melanoma. AB - Surgical excision of cutaneous malignant melanoma metastases is practical only when the number of lesions is small. In some patients isolated limb perfusion is not possible or fails to achieve control, and carbon dioxide laser ablation is then an alternative treatment. Between September 1992 and September 1994, 19 patients aged 45-94 years were treated with carbon dioxide laser. Two patients had received no previous limb perfusion, while the remaining 17 had up to three previous perfusions. The number of lesions per patient ranged from three to 40 nodules. A Sharplan 1030 portable carbon dioxide laser was used, which generates a beam wavelength of 10.6 microns and has a maximum power output of 80 W. All procedures were carried out under general anaesthesia. Each lesion was destroyed with a focused laser beam of 10-20 W with a spot size of 0.5-1.0 mm. At a mean follow-up of 15 months, five patients have died from the disease. Among the 14 survivors, eight have had no limb recurrence of the disease, three have had one further treatment and three a further two treatments to control cutaneous metastases at new sites. Early experience suggests that carbon dioxide laser ablation of cutaneous metastases is an effective palliative treatment after failed isolated limb perfusion, and there may be a group of patients in whom laser ablation should be the initial treatment of choice. PMID- 7489162 TI - Use of lightguide spectrophotometry to quantify skin oxygenation in a variable model of venous hypertension. AB - A variable model of venous hypertension was used to examine the immediate effect on skin oxygenation and blood flow. Measurements were carried out using laser Doppler flowmetry, transcutaneous oximetry and both macro- and micro-lightguide spectrophotometry to measure haemoglobin saturation (SO2). Assessments were performed on 20 normal subjects and in ten patients with deep venous insufficiency lying supine with a pressure cuff left uninflated (P0), then inflated to 40 mmHg (P40), 60 mmHg (P60) and 80 mmHg (P80). All measurements were performed at the gaiter area of the leg. In normal subjects median Doppler values fell significantly at P40 (P < 0.001) but further pressure increases had no more effect. Mean values of transcutaneous partial pressure of oxygen (PtcO2) fell steadily (P < 0.001), although the initial fall at P40 was small. Median SO2 fell with increasing occlusion pressure with both macro- and micro-lightguide spectrophotometry (P < 0.001), although the main reduction occurred predominantly at lower occlusion pressures. Higher levels of SO2 were obtained with local heating up to 44 degrees C, with a pattern similar to that of PtcO2. In patients, the same pattern of response to increased venous pressure occurred in areas of clinically normal skin. These data suggest that small increases in venous hypertension have an immediate and profound influence on skin blood flow and oxygen availability in normal skin. PMID- 7489161 TI - Presentation of endometriosis to general surgeons: a 10-year experience. AB - The presentation and management of 24 patients with endometriosis (median age 34 (range 21-68)years) presenting to general surgeons over a period of 10 years (1985-1994) was reviewed. Patients presented with an abdominal wall swelling related to a previous Pfannenstiel incision (seven patients), umbilical swelling (four), inguinal canal swelling (two), incidentally following appendicectomy (five), terminal ileal obstruction (two), rectal bleeding (two) and urinary symptoms (two). Endometriosis was not suspected in most patients but was confirmed by surgical excision or resection with minimal morbidity. No recurrence occurred during a median follow-up of 53 (range 9-113) months. Endometriosis is a disease rarely seen by general surgeons and is often diagnosed incidentally or on histological examination. Cyclical symptoms associated with menstruation are present in 50 per cent of patients and should suggest the diagnosis in those presenting with scar-related and/or subcutaneous swellings. Simple excision or resection of the presenting lesion provides adequate treatment but, since pelvic endometriosis may be present, referral to a gynaecologist is recommended in every case. PMID- 7489163 TI - Aspirin improves the outcome of intra-arterial thrombolysis with tissue plasminogen activator. Thrombolysis Study Group. PMID- 7489164 TI - Use of a semipermeable polyamide dressing over skin grafts to venous leg ulcers. AB - A polyamide semipermeable mesh interface placed over a skin graft allows absorption of exudate and soaking of the graft with an antiseptic solution. Forty five patients with large venous leg ulcers (more than 100 cm2) were treated by a split skin graft covered by a polyamide mesh. The mean time to healing and graft take was compared with those of a similar group of 30 patients whose graft was not protected by an interface. The mesh significantly shortened the mean time for complete healing from 41 to 32 days. PMID- 7489165 TI - Critical limb ischaemia in patients over 80 years of age: options in a district general hospital. AB - A total of 114 reconstructions were performed in 82 octogenarian patients and the results compared with those of 33 patients who had primary amputation. The operative mortality rate was higher after amputation (45 versus 11 per cent) and the mean survival less (25 versus 34 months). Quality of life assessment using a Rosser scale suggested that, although there was no improvement in the 38 per cent with a failed reconstruction, there was a significant improvement in the 62 per cent whose reconstruction remained patent. There was minimal improvement in quality of life after primary amputation and this was due to relief of pain. Costs (including the costs of revisions and community costs) were assessed in detail. Although the mean total operative costs of reconstruction were higher than those of amputation (10,222 pounds versus 6475 pounds) this was more than offset by the high community costs of amputation. The total cost of reconstruction was 13,546 pounds, compared with 33,095 pounds for amputation. Following reconstruction 66 per cent of those patients independent before critical limb ischaemia occurred were able to return to their own home; only 33 per cent of amputees were able to do so. PMID- 7489167 TI - Optimum method for urinary drainage in major abdominal surgery: a prospective randomized trial of suprapubic versus urethral catheterization. AB - The outcome of suprapubic and urethral catheterization in abdominal surgery was compared in a prospective randomized trial. Twenty-eight patients received a suprapubic and 29 a urethral catheter. The groups were similar in terms of age, sex, operation performed and postoperative analgesia. There was no difference in the duration of catheterization (suprapubic: median 5 (range 4-10) days; urethral: median 4 (range 2-11) days). Urinary sepsis occurred in three patients in each group. Urethral catheters caused pain in significantly more patients (urethral 13; suprapubic two; chi 2 = 8.6, 1 d.f. P < 0.01), on more days (suprapubic: 6 of 142 catheter days; urethral: 37 of 126 catheter days; chi 2 = 29.5, 1 d.f. P < 0.001). Two men with urethral catheters and one with a suprapubic catheter failed to void urethrally when required to do so. Suprapubic catheterization is the method of choice for urinary drainage when this is required in abdominal surgery. PMID- 7489168 TI - Prospective study of Helicobacter pylori infection in primary gastric lymphoma. PMID- 7489169 TI - Prospective evaluation of ultrasonography and liver function tests for preoperative assessment of the bile duct. AB - Current means of predicting the presence of bile duct stones are sensitive but produce a large number of false-positive results, potentially leading to added morbidity from unnecessary invasive investigation. This study prospectively assessed 545 patients with gallbladder stones, including 55 patients with bile duct stones, to refine criteria for further investigation of the bile duct. The sensitivity, specificity, and positive and negative predictive values were calculated for bile duct dilatation (diameter abnormal if larger than 5 mm below age 50 years, normal value increasing by 1 mm per decade) and elevation of three liver function tests, singly or in combination. Depending on the number of abnormalities used the sensitivity ranged from 46 to 96 per cent and the positive predictive value from 35 to 77 per cent. All groupings gave at least 96 per cent negative predictive value for the absence of stones but none combined both high sensitivity and positive predictive value. The specificity was improved if the tests were done on the working day before operation. Refining criteria for predicting bile duct stones allows the selective use of cholangiography and may reduce the number of negative cholangiograms. PMID- 7489166 TI - Lobular carcinoma of the breast can be managed by breast-conserving therapy. AB - There has been a reluctance to treat patients with invasive lobular carcinoma by breast-conserving therapy because of the possibility of increased rates of local recurrence. This study is a retrospective review of 226 patients with pure invasive lobular carcinoma 4 cm or less treated by breast conservation therapy or mastectomy. Local relapse occurred in four of 52 patients (8 per cent) after breast conservation compared with 21 of 174 (12 per cent) after mastectomy. Breast conservation for invasive lobular carcinoma did not result in a higher incidence of local relapse and patients with invasive lobular and ductal carcinomas may be managed in a similar fashion. PMID- 7489170 TI - Randomized trial of laparoscopic cholecystectomy and mini-cholecystectomy. AB - Three hundred and ten patients having elective cholecystectomy were randomized to either laparoscopic cholecystectomy or mini-cholecystectomy. There were 155 patients in each group. Conversion to open cholecystectomy was significantly more common with laparoscopic cholecystectomy (13 versus 4 per cent) and complications were significantly more frequent with laparoscopic cholecystectomy (9 versus 3 per cent). If laparoscopic cholecystectomy was successful, hospital stay was significantly shorter than for mini-cholecystectomy (2 versus 3 days respectively), but overall the hospital stay was not significantly different. Postoperative analgesia requirements were reduced and return to normal activities and to work were faster after laparoscopic cholecystectomy. There was no significant cost difference between the two procedures. PMID- 7489171 TI - Symptomatic outcome 1 year after laparoscopic and minilaparotomy cholecystectomy: a randomized trial. AB - In a randomized controlled trial, 299 patients were sent a symptoms questionnaire 1 year after laparoscopic (n = 151) or minilaparotomy (n = 148) cholecystectomy for symptomatic cholelithiasis. The response rate to the questionnaire from contactable patients was 86 per cent. In both groups, at least 90 per cent of patients reported that their symptoms were improved, and at least 93 per cent rated the success of their operation as 'excellent', 'good', or 'fair'. However, over half the patients reported abdominal pain, a quarter reported flatulence, and a quarter dyspepsia. The only difference between treatment groups was that a higher proportion of patients who underwent minilaparotomy reported heartburn (35 per cent versus 19 per cent, P = 0.005). Patients who reported a 'poor' outcome were more likely to have suffered a postoperative complication, had lower quality of life scores, and higher anxiety and depression scores. Both laparoscopic and minilaparotomy cholecystectomy result in symptomatic benefit in at least 90 per cent of patients with symptomatic cholelithiasis. PMID- 7489172 TI - Laparoscopic 'bikini mesh' repair of bilateral inguinal hernia. AB - Laparoscopic hernia repair using a single piece of mesh was performed in 150 patients with bilateral inguinal hernia. The median operating time was 43 (range 30-90) min with a median hospital stay of 1 (range 1-10) days. In all, 138 patients were discharged within 24 h of operation. The median time for return to normal activity was 7 (range 2-60) days and that for return to work 14 (range 2 60) days. One patient required surgery for a port-site hernia and another for a Veress needle injury to the small bowel. Additional complications included bruising in nine patients, cord seromas in seven and urinary retention in two. There have been no recurrences after a median follow-up of 18 (range 1-38) months. The cost benefits of a short hospital stay and rapid return to work afforded by laparoscopic bilateral hernia repair warrant further evaluation. PMID- 7489173 TI - The 554 ileostomy. PMID- 7489174 TI - Radical resection of locally advanced colorectal cancer. AB - Multivisceral resection has been accepted as treatment for patients with locally advanced colorectal cancer. Nodal status has recently been claimed to be the most important predictor of survival in patients with such disease, with no survival after 2 years for patients with lymph node metastasis. A retrospective analysis was carried out of the prognostic significance of different tumour characteristics, and whether a more limited palliative resection is warranted in patients with positive lymph nodes. Of 1346 patients with colorectal adenocarcinoma operated on between 1987 and 1991, all those with a tumour staged as T4N0M0 or T4N1M0 (94 patients) were selected. From the remainder, 195 patients with stage T3N0M0 and T3N1M0 lesions were randomly selected as a control group. Overall survival was assessed at the beginning of 1993. The most important predictors of survival were lymph node status and involvement of the resection margins of the tumour. Overall survival in patients with T4 tumours who underwent radical resection was not significantly different from that in those with T3 tumours, even in N1 stages. Extended resection did not induce unacceptable morbidity or mortality. Surgery for locally advanced colorectal adenocarcinoma should result in tumour-free margins, and should therefore include multivisceral resection, even in patients with lymph node metastasis. PMID- 7489175 TI - Clinical and physiological evaluation of anorectal eversion during restorative proctocolectomy. AB - Fifty patients had a restorative proctocolectomy with stapled end-to-end ileoanal anastomosis by the eversion technique. Median (interquartile range) maximum resting anal pressure was 90 (73-116) cmH2O before restorative proctocolectomy and 71 (51-88) cmH2O 1 year after surgery (P < 0.001). Median maximum squeeze pressure was 141 (110-185) cmH2O before surgery and 146 (118-186) cmH2O 1 year after surgery (P not significant). Median thresholds for sensation in the lower third of the anal canal before and 1 year after surgery were 5.3 and 7.1 mA, respectively (P = 0.006). One year after restorative proctocolectomy, all patients were continent, although two experienced leakage of mucus requiring a pad. Forty-two patients (84 per cent) could discriminate between faeces and flatus. Eversion of the anorectum during restorative proctocolectomy impairs the motor and sensory functions of the anal sphincter. Most patients achieved satisfactory anal continence, however, despite these physiological changes. PMID- 7489176 TI - Parastomal hernia: local resiting and mesh repair. PMID- 7489178 TI - Prognostic factors in surgery for local recurrence of rectal cancer. AB - Fifty-five patients had resection of locally recurrent rectal cancer. Fourteen patients (25 per cent) had distant metastases, which were resected concurrently in six (11 per cent). Thirty-three patients (60 per cent) had preoperative (one patient) or postoperative (32) external beam radiotherapy (45-60 Gy). The 5-year survival rate was 18 per cent with a median survival of 24 months. The median symptom-free interval was 24 months. At a median follow-up of 28 months 53 per cent of patients had a second local recurrence and 24 per cent metastases only. Treatment complications occurred in 12 patients (22 per cent), three (5 per cent) of whom died 3-10 months after operation. Variables that were significantly related with longer survival and palliation were the radical nature of the operation, the absence of severe symptoms (such as pain, obstruction or sepsis), a recurrent tumour diameter of less than 5 cm measured on the resected specimen and a normal carcinoembryonic antigen level after reoperation. A Cox regression model showed that recurrent tumour diameter was the only independent prognostic variable. Surgery for local recurrence achieved local control in 47 per cent of patients with a low morbidity and mortality rate. PMID- 7489177 TI - Prospective study of prognostic factors in patients with unresected hepatic metastases from colorectal cancer. Fondation Francaise de Cancerologie Digestive. AB - Prognostic factors of unresected liver metastases in patients with colorectal cancer are not well established. A total of 544 patients with unresected liver metastases from colorectal cancer were registered in a national survey over a 1 year period and followed until death. Twenty factors were studied in a univariate analysis (log rank test) and 16 influenced survival (P < 0.01). These 16 factors were entered in a multivariate analysis (Cox model) and eight, ranging from the most significant (relative risk of death 1.9) to the less significant (relative risk of death 1.2), independently influenced survival: performance status, alkaline phosphatase level, number of involved liver segments, administration of chemotherapy, presence of extrahepatic metastases, site of the primary tumour, prothrombin time and resection of the primary lesion. Two simple classifications are proposed, taking into account the performance status and the alkaline phosphatase level, or the performance status and the number of involved liver segments. PMID- 7489179 TI - High morbidity rate after converted laparoscopic colorectal surgery. AB - Sixteen of 65 laparoscopically assisted colorectal operations were converted to an open procedure, usually because of dissection difficulties. Conversion was decided early in the procedure in four cases without perioperative morbidity and later in 12 cases, after a mean of 56 min. The results in patients undergoing converted operation were compared with those in 252 having a planned open colorectal procedure during the same period. The groups were comparable with regard to age, health status, factors influencing intestinal healing and grade of surgeon. A higher postoperative morbidity rate (50 versus 21 per cent) and more anastomotic leakages (25 versus 8 per cent) were apparent in the converted group. Operating time, postoperative ileus and hospital stay were longer in those requiring a converted operation. These poor results suggest that careful preoperative patient selection for laparoscopic procedures and a rapid decision to convert in case of difficulty are important. PMID- 7489180 TI - Role of total pancreatectomy in the treatment of patients with end-stage chronic pancreatitis. AB - Forty patients underwent total pancreatoduodenectomy for end-stage chronic pancreatitis. There were 34 men and six women of median age 39 (range 21-66) years. Alcoholism was the major aetiological agent (30 patients), and five patients had had previous acute idiopathic pancreatitis. The overwhelming indication for operation was severe abdominal pain, complicated by failing exocrine and endocrine function. Resection was performed in one (17 patients) or two (23) stages, following previous proximal (seven) or distal (16) pancreatectomy; progression from partial to total pancreatectomy occurred over an interval of 8-96 (median 15) months. A further six patients had undergone previous pseudocyst or duct drainage procedures. The pylorus was preserved in 28 patients and the spleen in ten. Median operating time was 6 (range 2.5-8.5) h and median blood loss 2000 (range 500-16,000) ml. There were two hospital deaths and three patients required reoperation. Of 38 survivors, 30 obtained complete or substantial relief of pain. There were 15 late deaths at 2.5-120.0 months after operation, 13 in the alcohol group and 11 disease-related. Total pancreatectomy can relieve the intractable pain of chronic pancreatitis at the cost of possible premature death from continuing alcohol abuse. PMID- 7489181 TI - Acute intestinal ischaemia after laparoscopic cholecystectomy. PMID- 7489182 TI - Randomized, double-blind phase II trial of Lexipafant, a platelet-activating factor antagonist, in human acute pancreatitis. AB - The aims of the study were to determine whether the platelet-activating factor antagonist Lexipafant could alter the clinical course and suppress the inflammatory response of human acute pancreatitis. In a double-blind, placebo controlled study 83 patients were randomized to receive Lexipafant 60 mg intravenously for 3 days, or placebo. Clinical progression was assessed by daily Acute Physiology And Chronic Health Evaluation (APACHE) II score and organ failure score (OFS). The magnitude of the inflammatory response on days 1-5 was assessed by serial measurement of interleukin (IL) 8, IL-6, E-selectin, polymorphonuclear elastase-alpha1-antitrypsin (PMNE-alpha 1-AT), and C-reactive protein (CRP). At entry, patients receiving Lexipafant (n = 42) or placebo (n = 41) were matched for age and sex, aetiology, APACHE II score and OFS. The disease was classified as severe in 29 patients (APACHE II score eight or more). There was a significant reduction in the incidence of organ failure (P = 0.041) and in total OFS (P = 0.048) at the end of medication (72 h). During this time seven of 12 patients with severe acute pancreatitis who had Lexipafant recovered from an organ failure; only two of 11 with severe acute pancreatitis who had placebo recovered from an organ failure and two others developed new organ failure. Lexipafant treatment significantly reduced serum IL-8 (P = 0.038), and IL-6 declined on day 1. Plasma PMNE-alpha 1-AT complexes peaked on day 1; the gradual fall to baseline over 5 days observed in controls did not occur in patients given Lexipafant. No effect was observed on serum CRP. This study provides a rationale for further clinical trials with the potent PAF antagonist Lexipafant in human acute pancreatitis. PMID- 7489183 TI - Selective balloon occlusion of the portal tree through the umbilical vein for hepatic resection. PMID- 7489184 TI - Instrument safety during laparoscopic cholecystectomy. PMID- 7489186 TI - Prospective randomized study of radical versus four piles haemorrhoidectomy for symptomatic large circumferential prolapsed piles. PMID- 7489185 TI - Mortality, endotoxaemia and cytokine expression after intermittent and continuous hepatic ischaemia. AB - This study compared mortality rates, endotoxaemia, systemic tumour necrosis factor (TNF) and interleukin (IL)-6 concentrations after continuous and intermittent hepatic ischaemia. Two groups of rats were subjected to continuous or intermittent left hepatic inflow occlusion for a total period of 120 min in each group. Intermittent ischaemia was associated with significantly lower mortality rates than continuous ischaemia (four of 20 versus 15 of 20; P = 0.0015). In a separate study, again following 120 min continuous or intermittent ischaemia, systemic blood was sampled at 0 min, 1 h, 3 h and 5 h after final clamp release for measurement of endotoxin, TNF and IL-6 concentrations. Endotoxin concentrations were significantly lower at 1 h, as were TNF and IL-6 concentrations at 3 and 5 h, after final clamp release in the group having intermittent ischaemia (P < 0.05). Intermittent ischaemia is associated therefore with significantly reduced mortality rates and lower systemic endotoxin, TNF and IL-6 concentrations when compared with continuous ischaemia. PMID- 7489187 TI - Failure of macrophage activation in experimental obstructive jaundice: association with bacterial translocation. PMID- 7489188 TI - Preoperative endoscopic sphincterotomy during laparoscopic cholecystectomy for choledocholithiasis. PMID- 7489190 TI - Paraoesophageal hiatus hernia: an important complication of laparoscopic Nissen fundoplication. PMID- 7489189 TI - Peroperative endoscopic sphincterotomy during laparoscopic cholecystectomy for choledocholithiasis. PMID- 7489191 TI - Interest groups. PMID- 7489192 TI - Commentary on standards for media fill testing. PMID- 7489193 TI - PDA response: electronic records; electronic signatures. Parenteral Drug Association. PMID- 7489194 TI - The potential for use of steam at atmospheric pressure to decontaminate or sterilize parenteral filling lines incorporating barrier isolation technology. AB - Barrier isolators that enclose aseptic filling equipment are being proposed as a means of: (1) assisting in achieving a 10(-6) sterility assurance level (SAL) in the filling area and (2) minimizing the clean environment required in the manufacturing area. The need for operator and maintenance access to the interior of the barrier isolators presents difficulties in achieving the above goals. Several methods are available for reducing the microbial level inside the isolation barrier. If the objective is the decontamination of all surfaces inside the enclosure, saturated steam at atmospheric pressure can be used. If the objective is to sterilize the inside of the enclosure, saturated steam at atmospheric pressure with added H2O2 can be used. Test data and practical interface considerations relative to various methodologies will be reviewed. PMID- 7489195 TI - Comparison of D-value accuracy among Limited Spearman-Karber Procedure, Stumbo Murphy Cochran Procedure (SMCP) and Limited SMCP. AB - The Limited Spearman-Karber Procedure (LSKP) and Stumbo Murphy and Cochran Procedure (SMCP) are procedures to determine D-value in the Most Probable Number method (MPN). International Organization for Standardization (ISO)/Technical Committee 198/Working Group 4 contributing for the preparation of documents on biological indicator has adopted only LSKP in MPN. SMCP has been rejected due to the reason of inaccuracy without any scientific proof. Therefore, in order to confirm whether this is scientifically correct or not, Japanese delegates decide to compare both methods in simulation and experimental procedures using biological indicator and biological indicator evaluator resistometer. The result indicated that SMCP with some restrictions was not always inferior to LSKP and gave identical results. PMID- 7489196 TI - Environmental microbial challenges to an aseptic Blow-Fill-Seal process--a practical study. AB - A Blow-Fill-Seal machine, set to aseptically fill sterile liquid TSB medium into 20 ml plastic containers, was subjected to a series of environmental microbial challenges in order to evaluate the effects of the room environment, and of the actions of persons in the room, on the sterility of the product. Despite a series of relatively severe stresses to the system, none of the units filled (approximately 44,000) were found, following incubation, to display growth. PMID- 7489197 TI - Heat resistance of Bacillus coagulans spores suspended in various parenteral solutions. AB - Bacillus coagulans, ATCC 51232 (formerly referred to as FRR B666), was previously reported to be a potentially excellent biological indicator. The organism was grown on a relatively simple sporulation media and the spores were evaluated for moist heat resistance in various parenteral solutions. Resistance was determined using the survivor curve method. For those solutions which appeared to be inhibitory to the outgrowth of Bacillus coagulans, ATCC 51232, further investigations were performed to evaluate the inhibitory effect. The results obtained demonstrated that the organism had the highest resistance in Calcium Gluconate Injection; USP (100 mg/mL) at 120 degrees C. PMID- 7489199 TI - Opportunities and obstacles in the implementation of barrier technology. AB - This paper was developed from the keynote address given at the Isolation Technology Users Group (ITUG) meeting held in Raleigh, North Carolina in December of 1993. Present at the meeting were over 100 individuals representing a cross section of users and suppliers. Unfortunately not all suppliers of isolation technology equipment were represented at this meeting. Subsequent to the meeting, ITUG has decided to operate as a special interest group within the Parenteral Drug Association and attendance will be open to all PDA members. I anticipate that the affiliation of ITUG within PDA will facilitate the free exchange of ideas and information among those interested in barrier technology without interference from commercial considerations. My hope is that we can nurture within the industry a common effort among all users and suppliers to overcome the obstacles and realize the opportunities inherent in isolation technology. While it could be construed that certain of my comments represent an endorsement or a chastisement of certain suppliers of isolator systems that was certainly not my intent. It was prepared in a spirit of egalitarianism to assist the industry at large with what I believe to be a technology of great promise. PMID- 7489198 TI - Hazard analyses of airborne contamination in clean rooms--application of a method for limitation of risks. AB - Hazard Analyses and establishing of Critical Control Points (HACCP) using a method with Limitation of Risks, known as the LR Method are described. This method has a concept involving visualization of air movements, particle challenge testing and calculation of a risk factor. The calculated risk factor and the estimated exposure time form the basis for the HACCP. The LR Method can also be used as an engineering tool to achieve microbiologically safe processes with regard to airborne contamination. As an alternative to the increasing extent of microbiological monitoring sampling, a system is discussed that combines continuous monitoring of particle levels with particle counters and a limited monitoring of viable particles with air samplers. PMID- 7489200 TI - Validation, risk-benefit analysis. PMID- 7489201 TI - Automated method for determining Instron Residual Seal Force of glass vial/rubber closure systems. Part II. 13 mm vials. AB - Instron Residual Seal Force (IRSF) of 13 mm glass vial/rubber closure systems was determined using an Instron 4501 Materials Testing System and computerized data analysis. A series of three cap anvils varying in shape and dimensions were machined to optimize cap anvil performance. Cap anvils with spherical top surfaces and narrow internal dimensions produced uniform stress-deformation curves from which precise IRSF values were derived. PMID- 7489203 TI - Validation of computer-related systems. Parenteral Drug Association. PMID- 7489202 TI - Expert systems for parenteral development. AB - This article introduces and reviews the use of expert systems in parenteral development. Two case studies are reviewed, one from academia the other from industry. Where introduced and implemented, expert systems have generated significant benefits in terms of knowledge protection, cost reduction, training, consistency and improved communication. PMID- 7489204 TI - A review of the management of vertebral artery injury. PMID- 7489205 TI - Anti phospholipid syndrome. PMID- 7489206 TI - Operative indication in asymptomatic carotid arterial disease. PMID- 7489207 TI - Calf deep venous thrombosis: a review of the literature. PMID- 7489208 TI - Avidin and 111In-labelled biotin scan: a new radioisotopic method for localising vascular graft infection. AB - OBJECTIVES: To evaluate a new imaging technique, for diagnosis of prosthetic vascular graft infection. Avidin is a protein which accumulates nonspecifically at sites of inflammation or infection. Due to its extremely low dissociation constant with biotin sites of infection can be imaged, using avidin as a pre target, followed by injection of 111In-labelled biotin. This technique is much simpler than the common scintigraphic methods which employ labelling of blood components and its target-to-background ratio is greater than the methods employing radiolabelled proteins. DESIGN: Prospective clinical study. SETTING: A single department of vascular surgery and one of nuclear medicine of a Northern Italian hospital. MATERIALS: Between May 1993 and May 1994, 31 grafts in 26 patients were studied; the series included 23 men and three women with a mean age of 65.5 years (range 54-76 years). The prosthetic graft (Dacron -16, ePTFE -15) were: aortoaortic 5, aortobifemoral 15, aortoiliac 1, and femoropopliteal 10. Sixteen patients were suspected of having a vascular graft infection (Group A), the other 10 patients served as controls (Group B). 20 mg of Avidin were injected iv, followed 24 h later by i.v. injection of 500 micrograms of Biotin labelled with 74 MBq of 111In. CHIEF OUTCOME MEASURES: Whole-body imaging was performed at 10 min and 2 h post-injection, along with SPECT imaging when indicated. Scan results were correlated with the traditional imaging modalities and the clinical outcome of the patients. MAIN RESULTS: In Group A: two patients (three grafts) were excluded from the study, there were six true-positives, one false-positive and 11 true-negatives. Results in Group B: 10 true-negatives. The overall sensitivity was 100%, the specificity 95%, the accuracy 96%, the positive predictive value 86% and the negative predictive value 100%. CONCLUSIONS: These data suggest that Avidin/111In-labelled Biotin scintigraphy is a useful non invasive diagnostic method for early diagnosis of suspected prosthetic vascular graft infection. PMID- 7489209 TI - Compliance mismatch and formation of distal anastomotic intimal hyperplasia in externally stiffened and lumen-adapted venous grafts. AB - OBJECTIVE: Compliance and formation of distal anastomotic intimal hyperplasia (DAIH) were investigated in externally stiffened venous grafts of varying calibers. METHODS: 36 femoropopliteal reconstructions were performed in 18 sheep. The autologous venous grafts were inserted into tubes made of Dacron mesh to achieve compliance-mismatch and lumen adaptation. Compliance was measured by echotracked ultrasonography and profiles of DAIH were generated from histologic sections harvested after 8.3 months. MAIN RESULTS: The external mesh tube significantly lowered the local compliance of graft and host artery. DAIH appeared extensively in those groups where mesh tube constricted venous grafts met untreated host arteries (p = 0.002). No differences in compliance and DAIH formation were observed when grafts with large and adapted diameters were compared. CONCLUSIONS: For prevention of DAIH the distal venous graft diameter is not important, while the local compliance of an autologous vein is a predictive factor for DAIH formation and thus long-term patency. PMID- 7489210 TI - The role of the circle of Willis in carotid occlusion: assessment with phase contrast MR angiography and transcranial duplex. AB - PURPOSE: To study the collateral pathways recruited after occlusion of the internal carotid artery (ICA), and to evaluate its influence on the impairment of hemispheric blood flow supply and development of low flow infarcts. METHODS: 38 patients with ICA occlusion (18 asymptomatic; five transient ischaemic attacks; and 15 strokes) were included. Infarcts on cerebral MR scanning were categorised in order to differentiate patients with territorial infarcts or no lesion (group I; n = 22) from those with brain damage due to low flow (group II; n = 16). Patency and direction of flow in the communicating arteries were assessed by means of cine phase contrast MR angiography (PC-MRA). Flow velocity in the middle cerebral artery (MCA) was measured by means of transcranial Duplex (TCD). RESULTS: Cine PC-MRA revealed a reversed ophthalmic artery blood flow ipsilateral to the ICA occlusion in all except two patients in group I and one patient in group II (NS). Posterior to anterior flow in the ipsilateral posterior communicating artery (PCoA) was detected in 16 (73%) patients in group I and in 13 (81%) in group II (NS). In contrast, reversed blood flow in the ipsilateral A1 segment of the anterior cerebral artery, through a patent anterior communicating artery (ACoA), was identified in 19 (86%) patients of group I, vs. 7 (44%) of group II (p = 0.005). The relative risk of low-flow infarcts was significantly higher in those cases with non-functioning ACoA (odds ratio = 8.1; p < 0.05). TCD showed a lower peak systolic velocity (PSV) in the ipsilateral MCA than in the contralateral one (60 +/- 9 cm/s vs. 90 +/- 11 cm/s; p < 0.005). Those patients without crossed flow through the ACoA, showed an even lower PSV in the ipsilateral MCA (55 +/- 7 cm/s vs. 64 +/- 9 cm/s; p = 0.03). CONCLUSIONS: These data suggest that even though ICA occlusion may occur without cerebral damage, collateral blood supply is not enough to maintain normal hemispheric perfusion. The ACoA may be a key collateral pathway as a non-functioning ACoA is associated with an increased risk of developing low-flow infarcts. PMID- 7489213 TI - Cranial nerve injuries after carotid artery surgery. A prospective study of 663 operations. AB - OBJECTIVE: To establish the incidence of cranial nerve injuries caused by carotid artery surgery. METHODS: During the years 1982 to 1992, 689 carotid artery operations were performed at Malmo General Hospital. Of these 656 were pre- and postoperatively examined at the Department of Phoniatrics concerning the function of the cranial nerves IX, X, and XII. RESULTS: There was injury to one or more of the examined nerves after 75 operations (11.4%). In 70 (10.7%) operations the hypoglossal nerve was injured, in eight (1.2%) the recurrent laryngeal nerve, in two (0.3%) the glossopharyngeal nerve and in two (0.3%) the superior laryngeal nerve. Seven patients complained of postoperative hoarseness, but had no injuries. One hypoglossal and one recurrent nerve injury was permanent. Nerve injury was more frequent in operations performed with a shunt (p = 0.05), with patch closure (p = 0.01) and by a junior surgeon (p = 0.05). CONCLUSION: In a prospective series, the incidence of cranial nerve injuries after carotid surgery was significant but the vast majority were reversible. PMID- 7489212 TI - Durability of short bypasses to infragenicular arteries. AB - OBJECTIVES: The purpose was to test the durability of the use of the unobstructed popliteal or tibial arteries as alternative inflow sources. MATERIALS: We examined 106 such bypasses performed during a 12 year period (1981-93). The indication for surgery was limb salvage in 99.1%. Seventy-five percent of the patients were male, 78% were diabetic and the average age was 59.6 years. The inflow source was the above-knee popliteal artery in 15 cases, the below-knee popliteal artery in 70 cases, the anterior tibial artery in 11 cases and the posterior tibial artery in 10 cases. Adequacy of inflow was determined by angiogram as well as intraoperative pressure measurement when indicated. Outflow was to a distal tibial or plantar arteries in 77.4% of the procedures. Forty-six bypasses were performed by an in situ technique. Excised vein was used as conduit in 60 patients (56.6%); greater saphenous vein (38), lesser saphenous vein (10), cephalic/basilic vein (4), and splice vein (8). RESULTS: Operative mortality was 2.8%. Five year cumulative primary patency was 75.4% with a secondary patency of 82.6%. Five year cumulative limb salvage was 93.5%. Patency rate was not significantly different for various inflows or outflows. Only four of the 106 bypasses ultimately required a reconstruction from the femoral level for proximal progression of disease. Bypasses performed using an in situ technique showed a significantly better 5 year cumulative secondary patency rate (96.3%) than those done with excised vein (70.5%), p < .05 CONCLUSION: Results of this study indicate that use of the popliteal or tibial arteries as an inflow source in the absence of significant proximal disease carries acceptable results, especially when using the vein in situ. PMID- 7489211 TI - Morphology and function of dog arterial grafts preserved in UW-solution. AB - OBJECTIVES: To assess the function of arterial grafts after prolonged preservation in the University of Wisconsin solution (UW), in vitro and in vivo. METHODS: Carotid arteries were harvested from dogs and stored for 1-21 days at 4 degrees C in UW (n = 10) or in PBS (0.9% NaCl, pH 7.4), (PBS) (n = 10). Slices were examined by lightmicroscopy (LM) and scanning electron microscopy (SEM). For viability testing, specimens were connected to an isometric force transducer (2 x n = 9). Contractile and relaxation responses were examined by adding phenylephrine (200 microM) and metacholine (200 microM), respectively. For in vivo studies (n = 41), 2.5cm carotid artery segments were implanted or orthotopically, as autografts and allografts, after 14 days of storage in UW or in PBS. Autologous veins were used as controls. After 28 days or 56 days, arteriography was performed and the grafts were excised for LM and SEM. RESULTS: The arterial endothelial layer remained intact after up to 14 days of storage in UW. In PBS, the endothelium was lost after 3 days. The functional response after 14 days storage in UW was approximately 50% vs. 0% after 14 days in PBS. In the autografts, total patencies (28 days + 56 days) were 100% (8/8) and 63% (5/8) for UW and PBS stored grafts, respectively. In the allografts, the UW and PBS preserved grafts showed total patencies of 86% (12/14) and 83% (5/6), respectively. Microscopically, the allografts showed fibrotic degeneration. CONCLUSIONS: Arteries are well preserved in UW up to 14 days of storage. Arterial autografts preserved in UW showed good patency and better integrity of the vessel wall after implantation, than grafts stored in PBS or allografts (without immunosuppressive therapy). PMID- 7489214 TI - bFGF release is dependent on flow conditions in experimental vein grafts. AB - OBJECTIVES: Basic Fibroblastic Growth Factor (bFGF) is a powerful mitogen for smooth muscle cells and has been implicated in the genesis of Myointimal hyperplasia. The aim of this study was to determine the release of bFGF by veins in different haemodynamic conditions. DESIGN AND SETTING: Laboratory animal study. MATERIALS: In 39 Lewis rats, a 1 cm long segment of inferior vena cava was inserted at the level of the abdominal aorta. The segments of inferior vena cava were obtained from syngenic Lewis rats. Arterial Vein Grafts (AVG) were harvested after 4 weeks (AVG 4) and 12 weeks (AVG 12). In 16 animals the arterial vein grafts were explanted 4 weeks after the initial operation and reimplanted (Reimplanted Vein Grafts: RVG) in syngenic Lewis rats as venous-venous bypass grafts at the level of the left iliac vein and harvested after 2 weeks (RVG 2) and 8 weeks (AVG 8). OUTCOME MEASURES: The tissue was studied in organ culture in a serum-free system for (1) release of bFGF (immunoassay) and (2) mitogenic activity of the conditioned media. Scanning electron and light microscopy studies were also performed. RESULTS: bFGF release by veins increased significantly (p < 0.01) when veins were inserted in the arterial circulation, and decreased significantly (p < 0.01) when grafts were reimplanted in the venous system. bFGF release (ng/cm2): [Formula: see text] CONCLUSION: Vein inserted in the arterial circulation release a higher quantity of bFGF. This could explain in part, the formation of myointimal hyperplasia in arterial vein graft. PMID- 7489215 TI - Outcome and quality of life of patients with severe chronic limb ischaemia: a cohort study on the influence of diabetes. AB - OBJECTIVE: To determine the influence of diabetes on the use of arterial reconstruction, the rate of amputation and death, and the quality of life of patients with severe limb ischaemia. DESIGN: A prospective study of patients with the first episode of ischaemia. SETTING: University tertiary referral centre. METHODS: Thirty-seven patients with diabetes and 50 without diabetes, were studied over a 12 month period with complete follow-up. MAIN OUTCOME MEASURES: The proportion of patients undergoing an arterial reconstruction, amputation rate, death rate, and quality-of-life scores. RESULTS: Patients with diabetes underwent an arterial reconstruction less often than patients without diabetes (7/37 vs. 18/50). The odds of patients with diabetes having a higher incidence of adverse outcome was 1666:1 for minor amputation, 26:1 for major amputation, and 4.7:1 for death. There was a tendency towards a lower quality of life for patients with diabetes at 3 (OR 1.94, p = 0.036), 6 (OR 1.58, p = 0.117), and 12 (OR 1.47, p = 0.185) months. CONCLUSIONS: In patients with diabetes, (1) the opportunity of undergoing an arterial reconstruction is lower, (2) morbidity and mortality are higher, and (3) the quality of life tends to be worse. PMID- 7489216 TI - Decreased mortality of abdominal aortic aneurysms in a peripheral county. AB - OBJECTIVES: To analyse the effect on the mortality associated with abdominal aortic aneurysms, due to the establishment of a decentralised vascular surgical unit in the county of Viborg. METHODS: Death after aneurysm repair and from rupture without repair were analysed retrospectively for the 3 year period before (1986-88), and after (1989-91), the unit was established. RESULTS: Between 1986 and 1988, one patient (5%) died after 19 elective and three emergency non ruptured aneurysm repairs. Only three ruptures were repaired with two deaths. In the country, 41 deaths due to ruptured aneurysm were recorded. Between 1989 and 1991 two patients (4%) died after 26 elective and 27 emergency non-ruptured aneurysm repairs. Nineteen ruptures were repaired with six deaths (32%). In the county, 28 deaths due to rupture were recorded during this period. CONCLUSIONS: The establishment of a decentralised vascular surgical unit has increased the proportion of ruptured aneurysms reaching surgery with a resultant decrease in mortality from this condition. PMID- 7489217 TI - The potential of simple clinical information and electrocardiogram to predict mortality of primary elective abdominal aortic reconstruction. AB - OBJECTIVE: To assess the importance of simple clinical information to predict mortality after primary elective aortic reconstruction. DESIGN: Retrospective clinical study. MATERIALS: Four hundred-three consecutive patients (206 with aortic occlusive disease (51%) and 197 with aneurysmal disease of the abdominal aorta (49%) who underwent primary elective aortic reconstruction during an 8 year study period. METHODS: Eight potential clinical risk variables and six electrocardiogram related risk variables were studied. The predictive value for postoperative mortality was evaluated univariately for all separate variables by calculation of the odds ratio (OR). Determination of the most predictive sets of risk factors was performed by stepwise logistic regression. RESULTS: Five clinical risk factors showed significant odds ratio's: > 70 years (OR: 4.1), aortic aneurysm (OR: 9.0), myocardial infarction (OR: 8.6), angina pectoris (OR: 4.6), congestive heart failure (OR: 8.1), all p < 0.01. Furthermore, four electrocardiogram related factors showed significant odds ratio's: infarction pattern (OR: 4.3, p = 0.003), ischaemic changes (OR: 6.2, p < 0.001), conductance disturbances (OR: 3.2, p = 0.04), and non-sinus rhythm (OR: 6.2, p = 0.003). Stepwise logistic regression analysis demonstrated that 3 clinical risk factors (aneurysm (OR: 6.1, p = 0.02), myocardial infarction (OR: 3.7, p = 0.01), and congestive heart failure (OR: 5.3, p = 0.006)) contributed significantly to mortality. Addition of any of the electrocardiogram risk factors did not contribute to the prediction of mortality in the presence of these factors. CONCLUSIONS: The patients' history and clinical examination provides the most useful information for further selection of additional cardiac tests before elective primary aortic reconstruction. The additional value of the electrodiogram is somewhat overestimated. PMID- 7489218 TI - Design issues of a randomised controlled clinical trial on spinal cord stimulation in critical limb ischaemia. ESES Study Group. AB - OBJECTIVES: Review of the design of a clinical study to evaluate of the efficacy of epidural spinal cord electrical stimulation (ESES) as compared to best medical treatment in patients with nonreconstructible critical limb ischaemia. DESIGN: Randomised controlled clinical trial of pragmatic type, which will be analysed according to the intention-to-treat principle. The treatment strategies are ESES, in addition to best medical treatment, and best medical treatment alone. Patients are followed-up for at least 18 months. SETTING: The ESES-trial is an ongoing multicentre trial in 17 hospitals in The Netherlands. PATIENTS: Patients with critical limb ischaemia, nonsuitable for either primary intervention or reintervention after failing reconstructions. CHIEF OUTCOME MEASURES: Limb survival, patient survival, quality of life and cost-effectiveness. MAIN RESULTS: From November 1991 until May 1994 120 patients had been enrolled. Using life table analysis, at one year 76% of these randomised patients were alive: 41% without amputation and 35% with amputation. Quality of life of the trial patients was low, even compared to other severely ill patient groups, such as liver and heart transplant candidates. CONCLUSIONS: Considering the high incidence of death and amputation, 18 months of follow-up seems adequate to detect a clinically relevant outcome improvement from ESES-treatment, if present. We hope to present the results of this study at the end of 1995. PMID- 7489219 TI - Carotid duplex scanning: patterns of referral and outcome. AB - OBJECTIVE: Review of the results of carotid Duplex scanning. DESIGN: Retrospective review of new referrals for carotid Duplex scanning to a regional vascular laboratory. MATERIALS: 1041 referrals made over an 18 month period. METHODS: Referrals were reviewed for scan quality, diagnostic category and outcome of scanning. RESULTS: The overall detection rate of ipsilateral stenoses > 70% and occlusions was 13.5%; the detection rate of surgically significant carotid lesions (70-99%) was only 8%. Analysis by diagnostic category revealed a significantly greater detection rate for carotid lesions > 70% in patients presenting with amaurosis fugax (24%) in comparison to those with TIAs (9.5%, p = 0.0005) or following a stroke (13%, p = 0.01). Patients referred with asymptomatic bruits or prior to cardiac surgery showed a positive scan rate of 30%. Sixty-seven carotid endarterectomies have resulted from scans performed in this period. CONCLUSION: The overall rate of detection for surgically relevant lesions is low, but in comparison to a similar audit conducted in 1992, the absolute number of carotid lesions > 70% has risen by 60%. An 8% detection rate for surgically relevant lesions may be valuable in projecting the need for carotid endarterectomy arising from a given number of Duplex scans. PMID- 7489220 TI - A safe combined surgical/radiological approach to endoluminal graft stenting of a popliteal aneurysm. PMID- 7489221 TI - Successful resection of simultaneous aneurysms of the superior mesenteric and right renal artery. PMID- 7489222 TI - Functional obstruction of the subclavian vein caused by a traumatic dissection flap. PMID- 7489223 TI - Reperfusion injury tourniquet syndrome caused by acute aortic occlusion due to embolism following pneumonectomy. PMID- 7489224 TI - Superficial femoral artery aneurysm--an uncommon site of aneurysm formation. PMID- 7489225 TI - False aneurysm of the anterior tibial artery following removal of tibial plate. PMID- 7489226 TI - Quality of life. PMID- 7489227 TI - Quality of life. PMID- 7489228 TI - Distribution and correlates of plasma fibrinogen in middle-aged women. Initial findings of the Postmenopausal Estrogen/Progestin Interventions (PEPI) study. AB - Fibrinogen levels have been reported in cohort and case-control studies to be positively related to the development of coronary heart disease. This report presents the distribution and determinants of fibrinogen in women enrolling in a 3-year randomized trial of hormone replacement therapy (HRT), the Postmenopausal Estrogen/Progestin Interventions (PEPI) trial. Fasting plasma fibrinogen levels were measured in 874 postmenopausal women, aged 45 to 65 years, who had not used HRT for at least 3.5 months. Mean (+/- SD) fibrinogen level was 2.83 +/- 0.45 g/L. There was a significant positive association between fibrinogen and age (P = .03). Significantly higher (P < .005) fibrinogen levels were seen in current smokers versus nonsmokers (2.94 versus 2.81 g/L), in women who reported consuming fewer than 12 alcoholic drinks in the 12 months before the baseline visit versus higher consumption (2.90 versus 2.79 g/L), and in women who reported never versus ever having used HRT (2.90 versus 2.77 g/L). Self-reported leisure time physical activity (LTPA) was negatively associated (P = .0001) with fibrinogen levels as follows: inactive (2.84 g/L), light (2.89 g/L), moderate (2.80 g/L), and heavy (2.60 g/L), with significantly (P = .0001) lower levels in women who reported heavy LTPA versus each of the other categories and in women reporting moderate versus light LTPA. A strong positive correlation was found between fibrinogen and body mass index (BMI) (r = .32; P < .0001). In a model that included age, smoking, alcohol intake, prior HRT, LTPA, and BMI, LTPA was no longer a statistically significant predictor of fibrinogen level, while associations with other variables remained significant.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7489229 TI - Dominant social status and contraceptive hormone treatment inhibit atherogenesis in premenopausal monkeys. AB - The stress of social subordination is associated with exacerbation of coronary artery atherosclerosis in premenopausal cynomolgus monkeys, possibly as a result of the ovarian dysfunction that reliably accompanies subordinate social status. The primary objective of the current study was to determine whether treatment with an oral contraceptive (OC) provides relative protection from development of atherosclerotic plaques, especially among animals made vulnerable to atherosclerosis by social subordination. In the present study, 193 adult female monkeys (Macaca fascicularis) were placed in social groups of 5 or 6 animals each. Half of the animals were then fed an atherogenic diet to which had been added a triphasic OC, while the remainder received only the atherogenic diet. At the end of 26 months, atherosclerosis was measured in an iliac artery biopsy taken from each monkey. The results demonstrated that among untreated animals subordinate individuals developed significantly more atherosclerosis than did their dominant counterparts (P < .01); however, OC treatment inhibited atherosclerosis in subordinate animals (P < .05) and eliminated the difference between dominant and subordinate animals that was observed in the untreated condition. Subordinate social status and OC treatment were both associated with reduced plasma concentrations of HDL cholesterol (P < .01 for both), and subordinate monkeys also had elevations in LDL cholesterol plus VLDL cholesterol (P < .01). Nonetheless, the interaction between social status and OC treatment remained significant even after covariance adjustment for variation in plasma lipids. Taken together, these results suggest that social subordination worsens, whereas OC treatment inhibits, atherosclerosis, and that these effects are independent of concomitant variability in plasma lipids. PMID- 7489230 TI - Compared with dietary monounsaturated and saturated fat, polyunsaturated fat protects African green monkeys from coronary artery atherosclerosis. AB - Atherogenic diets enriched in saturated, n-6 polyunsaturated, and monounsaturated fatty acids were fed to African green monkeys for 5 years to define effects on plasma lipoproteins and coronary artery atherosclerosis. The monkeys fed polyunsaturated and monounsaturated fat had similar plasma concentrations of LDL cholesterol, and these values were significantly lower than for LDL in the animals fed saturated fat. Plasma HDL cholesterol concentrations were comparable in animals fed saturated and monounsaturated fat and were significantly higher than in animals fed polyunsaturated fat. Thus, the monounsaturated fat group had the lowest LDL/HDL ratio. LDL particle size was largest in the saturated and monounsaturated fat groups, significantly larger than in the polyunsaturated fat group. LDL particle enrichment with cholesteryl oleate was the greatest in the animals fed monounsaturated fat, next greatest in the saturated fat-fed animals, and was least in the polyunsaturated fat-fed animals. Coronary artery atherosclerosis as measured by intimal area was less in the polyunsaturated fat compared with the saturated fat groups, was less in the animals fed polyunsaturated fat compared with the monounsaturated fat-fed animals, but did not differ between the monounsaturated and saturated fat groups. Cholesteryl ester, particularly cholesteryl oleate, accumulation in the coronary arteries was also similar between groups fed monounsaturated and saturated fat but was minimal in the animals fed polyunsaturated fat. In sum, the monkeys fed monounsaturated fat developed equivalent amounts of coronary artery atherosclerosis as those fed saturated fat, but monkeys fed polyunsaturated fat developed less. The beneficial effects of the lower LDL and higher HDL in the animals fed monounsaturated fat apparently were offset by the atherogenic shifts in LDL particle composition. Dietary polyunsaturated fat appears to result in the least amount of coronary artery atherosclerosis because it prevents cholesteryl oleate accumulation in LDL and the coronary arteries in these primates. PMID- 7489231 TI - Influence of diets rich in saturated and omega-6 polyunsaturated fatty acids on the postprandial responses of apolipoproteins B-48, B-100, E, and lipids in triglyceride-rich lipoproteins. AB - The effects of diets rich in saturated fatty acids (SFA) (total polyunsaturated fatty acids [PUFA] [g]/total SFA [g][P/S ratio], 0.2) or omega-6 PUFA (P/S ratio, 1.3) on the postprandial response of triglyceride-rich lipoproteins (TRL) was determined in normolipidemic young men. After 15 and 29 days of diet intervention, the postabsorptive concentrations of apolipoprotein (apo) B-48 and apoB-100 were higher in the SFA group than in the PUFA group, but the absolute increase in apoB-48 was similar 3 hours after a challenge meal containing one third of daily energy and returned to postabsorptive values at 6 hours; this response was closely coupled to that of TRL triglycerides. In both groups, the percent increase in TRL apoB-48 and triglycerides was greater after the PUFA meal than after the SFA meal. The concentration of TRL apoB-100 also increased at 3 hours in both diet groups but returned to postabsorptive values at 6 hours only in those fed the PUFA diet; in the SFA group, apoB-100 remained high at 6 hours and fell below postabsorptive values only 9 hours after the meal. This apoB-100 response was affected primarily by the fatty acid composition of the diet and not by that of the challenge meal. The postprandial response of apoB-100 was closely coupled to that of cholesterol and apoE. These observations suggest that in healthy young men, neither the fatty acid composition of the diet nor that of the challenge meal affects the clearance of chylomicron remnants after a fat containing meal. By contrast, the postprandial accumulation of hepatogenous TRL is prolonged in individuals fed a diet rich in SFA. PMID- 7489232 TI - Association of postprandial triglyceride and retinyl palmitate responses with asymptomatic carotid artery atherosclerosis in middle-aged men and women. The Atherosclerosis Risk in Communities (ARIC) Study. AB - Blood lipid alterations after a fatty meal may be atherogenic, but there is little information regarding their associations with disease independent of fasting lipids. Asymptomatic atherosclerosis cases (n = 229) and 373 control subjects free of atherosclerosis, as defined by carotid intima-media thickness on ultrasound images, were given a fatty meal with vitamin A, followed by 3.5- and 8 hour measurements of triglycerides (TGs), TG-rich lipoprotein TGs, apoproteinB48, and retinyl palmitate. Among white men and women but not among blacks, case status was associated with greater postprandial responses of TGs and TG-rich lipoprotein TGs, but only in nonobese persons (body mass index < 30 kg/m2). The associations were strong and significant after controlling for coronary risk factors (odds ratio, approximately 2.0) and fasting TGs (odds ratio, 1.5). Associations with other postprandial lipid measurements did not persist after controlling for fasting lipids. Elevated postprandial TGs appear to be an independent risk factor for carotid intimal thickening in nonobese whites. The lack of such a relation in obese subjects and the lipid profile they manifest suggest that postprandial TGs must be accompanied by accumulation of TG-rich lipoprotein remnants to be atherogenic. PMID- 7489233 TI - Abnormal reverse cholesterol transport in controlled type II diabetic patients. Studies on fasting and postprandial LpA-I particles. AB - The high incidence and prevalence of coronary heart disease in diabetes mellitus is clearly established. The usual lipid pattern found in type II diabetic patients is a moderate increase in fasting triglyceride levels associated with low HDL cholesterol levels. These abnormalities are further amplified in the postprandial state. To study the effect of these alterations on reverse cholesterol transport, we isolated lipoprotein containing apoA-I but not apoA-II (LpA-I) particles by immunoaffinity chromatography from the plasma of well controlled type II diabetic patients and nondiabetic matched control subjects. Different parameters involved in this antiatherogenic pathway were measured in both fasting and postprandial states. Diabetic patients had reduced levels of LpA I particles that were protein enriched and phospholipid depleted. Gradient gel electrophoresis showed that control LpA-I particles had five distinct populations, whereas diabetic particles lacked the largest one. LpA-I isolated from diabetic plasma exhibited a decreased capacity to induce cholesterol efflux from Ob 1771 adipose cells both in fasting (15.1 +/- 10.0% versus 7.5 +/- 2.7%, P < .05) and postprandial (17.7 +/- 11.2% versus 7.7 +/- 3.9%, P < .05) states, whereas only control particles showed significantly higher ability to promote cholesterol efflux after the test meal (P = .02). Lecithin:cholesterol acyltransferase activity measured with an exogenous substrate showed a 54% increase and an 18% decrease postprandially for control subjects and patients, respectively. Thus, the different abnormalities found in the fasting state were further amplified in the postprandial situation. This resulted in LpA-I particles with aberrant size and composition and decreased ability to accomplish their antiatherogenic role in type II diabetic patients. PMID- 7489234 TI - LDL size and subclass pattern in Mexico City residents and San Antonio Mexican Americans. AB - Recent studies suggest that a relative abundance of small dense LDL is a risk factor for coronary heart disease. We compared LDL size in Mexico City residents (n = 191) and San Antonio Mexican Americans (n = 282), two genetically similar populations that differ markedly in dietary behaviors: in Mexico City approximately 62% of calories are from carbohydrate and approximately 19% from fat, and in San Antonio approximately 40% of calories are from carbohydrate and approximately 40% from fat. Mean LDL size in Mexico City was 258.6 +/- 0.9 A, and in San Antonio, 255.9 +/- 0.6 A (P = .013). After adjustment for the higher triglyceride and lower HDL cholesterol levels (the two most important predictors of LDL size) in Mexico City, LDL size was significantly lower in San Antonio than in Mexico City by -8.33 +/- 0.84 A (P < .001). Our data suggest that the higher triglyceride concentrations in Mexico City residents that are associated with a higher carbohydrate diet may not be associated with atherogenic changes in LDL. PMID- 7489236 TI - A prospective, randomized trial of phenytoin in nonepileptic subjects with reduced HDL cholesterol. AB - Observational studies have demonstrated a positive association between phenytoin use and HDL cholesterol (HDL-C). Our goal was to determine whether phenytoin raises HDL-C in nonepileptic subjects at risk for coronary artery disease. We performed a double-blind, placebo-controlled, parallel-group study in 41 subjects with reduced levels of HDL-C. Subjects were placed on an American Heart Association Step I diet and were randomized to receive either phenytoin or placebo for 3 months. Serum levels of phenytoin were monitored and adjusted to between 7.5 and 15 micrograms/mL. Fasting levels of lipids and lipoproteins were determined twice at baseline (weeks -2 and -1) and during the treatment phase of the study (weeks 11 and 12). Compared with dietary baseline, phenytoin-treated subjects experienced significant paired percent increases in total HDL-C (12.4%; P < .01), an effect confined to the HDL2 subfraction (137%; P < .01). The paired percent increases in HDL-C and HDL2 levels remained significant after adjustment for placebo (P < .05, P < .025, respectively). There were no significant differences in the paired percent changes from dietary baseline in total cholesterol, triglyceride, or LDL cholesterol levels between placebo and phenytoin-treated groups. The significant paired percent increases in total HDL-C and HDL2 from dietary baseline suggest a potential role for phenytoin in subjects with reduced levels of HDL-C. PMID- 7489235 TI - Effects of 1 year of growth hormone therapy on serum lipoprotein levels in growth hormone-deficient adults. Influence of gender and Apo(a) and ApoE phenotypes. AB - We investigated the influence of gender and apoE and apo(a) phenotypes as well as the effect of the metabolic effects of growth hormone (GH) on the effect of GH therapy on serum lipoprotein concentrations in GH-deficient (GHD) adults. Forty four consecutive patients, 30 men and 14 women aged 46.5 (range, 19 to 76) years with GHD due mainly to pituitary tumors, were treated with recombinant human GH for 12 months. Serum concentrations of lipoproteins, insulin, thyroxine, and insulin-like growth factor-I were determined, body composition was assessed by bioelectrical impedance, and apo(a) and apoE phenotypes were analyzed. Lipoprotein(a) [Lp(a)] concentrations in the GHD subjects were compared with a gender- and apo(a) phenotype-matched control group. After 12 months of GH treatment, the total cholesterol, LDL cholesterol, and apoB concentrations decreased, the HDL cholesterol and apoE concentrations increased, and the apoA-I and triglyceride concentrations were unchanged. Before treatment, the Lp(a) concentration was similar to that in the control group. However, after 12 months of treatment, the Lp(a) concentration had increased by 44% and 101% above baseline and the control group, respectively. Men and women responded differently to GH, with a more marked increase in Lp(a) concentration and fat-free mass and a more pronounced decrease in body-fat mass in men. Apo(a) phenotypes had no major influence on the effect of GH therapy. The only significant difference between apoE phenotypes was a higher baseline Lp(a) concentration among apoE4 heterozygotes.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7489237 TI - Apolipoprotein A-I deficiency. Biochemical and metabolic characteristics. AB - Familial HDL deficiencies are associated with variable susceptibility to premature coronary heart disease, but the mechanism underlying this association remains poorly understood. Three homozygotes with isolated complete apo A-I deficiency caused by an autosomal codominant apo A-I Q[-2]X mutation and one heterozygote developed coronary heart disease before age 40 years. We characterized the effects of this mutation on lipoprotein metabolism. LDL FC, phospholipid, and apo B were all significantly higher in homozygotes than in heterozygotes. The HDLs of the heterozygotes were apo A-I poor relative to apo A II. Lecithin-cholesterol acyltransferase activity was 59% lower in homozygotes than in normal subjects or heterozygotes. Cholesteryl ester transfer activity was increased in a homozygote compared with a normolipidemic control subject. Postprandial lipid metabolism was studied in one homozygote and one heterozygote. Post-prandial TG response in the homozygote was significantly exaggerated, while residual plasma HDL level remained unaffected. The homozygote also had delayed clearance of retinyl ester, a marker of chylomicron remnant metabolism. Thus, homozygosity and heterozygosity for apo A-I Q[-2]X are associated with qualitative, as well as quantitative, disturbances in plasma HDLs, LDLs, lipid modifying enzyme activities, and postprandial retinyl ester metabolism. The observed elevation of atherogenic lipoproteins and reduction in antiatherogenic lipoproteins in the affected members of the apo A-I Q[-2]X kindred are consistent with the primary deficiency in apo A-I having pleiotropic effects that markedly enhance susceptibility for coronary heart disease. PMID- 7489238 TI - Familial hypobetalipoproteinemia is not associated with low levels of lipoprotein(a). AB - To assess whether very low concentrations of LDL affected lipoprotein(a) [Lp(a)] concentrations and apo(a) associations with lipoproteins, we studied Lp(a) levels and associations in heterozygous subjects with familial hypobeta-lipoproteinemia FHBL) associated with several truncated forms of apoB-100, ranging from apoB-31 to apoB-89. Distributions of apo(a) isotypes were assessed by a combined electrophoresis-immunoblotting procedure that detects 34 isoforms. Lp(a) concentrations were quantified by two ELISAs, one detecting total apo(a) and the other apoB-bound apo(a) in plasma. Associations of apo(a) with plasma lipoproteins were evaluated by gel permeation chromatography (FPLC) and DGUC followed by analyses of elution and gradient fractions by apo(a) ELISA. In addition, associations were examined by nondenaturing electrophoresis or immunoprecipitation of whole plasma and examination of contents by immunoblotting. Finally, interactions between r-apo(a) and LDLs were evaluated in reconstitution experiments. The distributions of apo(a) isotypes did not differ between FHBL-affected and unaffected members of the same kindreds, and concentrations of Lp(a) were similar even when subjects were matched for isotypes both within and across kindreds. In subjects heterozygous for apo(a) isoforms, the smaller isoforms were inversely related to Lp(a) concentrations, the larger isoforms were not. The regression lines between Lp(a) concentrations and the smaller apo(a) isoforms were significant and negative in slope for both FHBL affected and unaffected subjects, but the slopes of the lines did not differ. In multiple regression analyses, only the sizes of the smaller apo(a) isoforms contributed to the prediction of Lp(a) concentrations. ApoB-size made no difference. In simple apoB-100/apoB-truncation heterozygotes, virtually all apo(a) was complexed with apoB-100-containing particles but not apoB-truncation particles, and r-apo(a) recombined with apoB-100-containing LDLs but not with apoB-89-containing LDLs. Thus, (1) low apoB levels do not affect the plasma concentrations of Lp(a), (2) apo(a) binds apoB-100 to form Lp(a) particles of usual sizes and densities, and (3) apoB truncations even as large as apoB-89 do not form covalent bands with apo(a), although noncovalent associations with apoB 89 may be present in plasma. PMID- 7489239 TI - Ten LDL receptor mutants explain one third of familial hypercholesterolemia in a German sample. AB - Mutational defects in the LDL receptor are responsible for familial hypercholesterolemia (FH); thus far more than 150 mutations have been described. Nevertheless, systematic searches among the Germans have not been conducted. We used single-strand conformational polymorphism and polymerase chain reaction to find mutations in 10 index patients and in 40 other individuals with heterozygous FH. Our screen in the 10 index patients revealed 7 hitherto undescribed mutations. A screen of the 40 additional FH patients disclosed 20 defective of 54 total alleles and allowed specific diagnoses in 88 family members. We also found two families in which the children were markedly affected by FH, but the expected parental expression of the trait was not manifest. This observation suggests a role for additional environmental and genetic influences. Our report represents the first comprehensive effort to identify FH mutations in Germany. We found 10 mutations and these mutations explain 37% of FH cases. Our data may have relevance to expected FH patterns in central Europe. PMID- 7489240 TI - Biochemical prototype for familial thrombosis. A study combining a functional protein C mutation and factor V Leiden. AB - Resistance to activated protein C (APC) is associated with a single amino acid substitution in factor V (Arg506-->Gln, factor V Leiden) that results in delayed inactivation of the molecule by APC. The mutation is present in 20% of patients with a first episode of deep venous thrombosis. Arterial and venous thromboses are also associated with the type II protein C deficiency (protein CVermont). In protein CVermont, the substitution Glu20-->Ala alone (rPC gamma 20A) is responsible for the defective anticoagulant properties of PCVermont. It was recently established that a thrombotic episode occurred in 73% of family members who are heterozygous for both a functional protein C gene mutation and the factor V Leiden mutation. We evaluated the molecular defect that would accrue in the combined deficiency state of factor VR506Q/VaR506Q and rAPC gamma 20A using recombinant APC and natural purified factor VR506Q from patients homozygous for the Arg506-->Gln substitution. While wild-type recombinant APC (rAPC) slowly cleaves and inactivates factor VR506Q and factor VaR506Q, minimal cleavage of membrane-bound factor VR506Q and VaR506Q by rAPC gamma 20A at Arg306 and Arg679 occurs, and no loss in cofactor activity is observed. Our data demonstrate that rAPC gamma 20A cannot inactivate either factor VR506Q or factor VaR506Q at biologically relevant rates because of impaired cleavage at Arg306 and Arg679.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7489241 TI - Effect of selective factor Xa inhibition on arterial thrombus formation triggered by tissue factor/factor VIIa or collagen in an ex vivo model of shear-dependent human thrombogenesis. AB - Tick anticoagulant peptide (TAP) is a potent and selective inhibitor of factor Xa. TAP has shown good antithrombotic efficacy in experimental animal models of disseminated intravascular coagulation and venous and arterial thrombogenesis. In the present study we evaluated the effect of recombinant TAP (rTAP) on acute thrombus formation in human nonanticoagulated blood triggered either by tissue factor (TF) or by collagen at arterial shear conditions. The main goal was to establish the role of factor Xa in thrombus formation by use of an optimal inhibitory concentration of rTAP. Blood was drawn directly from an antecubital vein by a pump over the respective thrombogenic surfaces, which were positioned in a parallel-plate perfusion chamber. rTAP was mixed homogeneously into the flowing blood by a heparin-coated device positioned proximal to the perfusion chamber. The passage of blood through this device caused minor activation of coagulation but little activation of platelets. Fibrinopeptide A and beta thromboglobulin levels after 5 minutes of blood perfusion were, on average, 14 ng/mL and 45 IU/mL, respectively. rTAP at a plasma concentration of 0.90 mumol/L completely inhibited TF/factor VIIa-dependent thrombus formation at wall shear rates of 650 and 2600 s-1. These shear conditions are comparable to those in medium-sized arteries and in moderately stenosed small arteries, respectively. In contrast to the TF-coated surface, rTAP was less efficient in reducing collagen induced thrombus formation. While a significant reduction of 53% was observed at 650 s-1, thrombus formation at 2600 s-1 was not affected by rTAP.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7489242 TI - A monoclonal antibody that recognizes the GPIIb/IIIa antagonist DMP 728. Reversal of the effects of DMP 728 on platelet aggregation and bleeding time in the dog. AB - Since hemorrhagic events represent a major safety concern associated with the use of new antithrombotic therapies such as glycoprotein (GP) IIb/IIIa receptor blockade, we evaluated the ability of a monoclonal antibody recognizing DMP 728 (cyclic [D-2-aminobutyryl-N2-methyl-L-argininyl-glycyl-L-aspartyl-3- aminomethyl benzoic acid] methanesulfonic acid salt), a potent GPIIb/IIIa receptor antagonist, to reverse the pharmacological actions of DMP 728 in the dog. DC11 was chosen for in vivo evaluation based on its ability to inhibit the binding of [3H]DMP 728 to activated platelets and to attenuate the inhibition of ADP-induced aggregation on platelet-rich plasma ex vivo by DMP 728. After anesthesia mongrel dogs were given DMP 728 (20 micrograms/kg body wt IV) infused into the femoral vein, bleeding times were determined using a Simplate device from incisions on the backside of the tongue, and platelet aggregation was determined ex vivo. Nearly complete inhibition of platelet aggregation was observed for the dogs treated with DMP 728 (20 ug/kg IV) for up to 210 minutes, and bleeding times were prolonged > 15 minutes for 2 hours and remained elevated for more than 4 hours. DC11 (0.2 or 1.0 mg/kg body wt IV) given to dogs 10 minutes after DMP 728 resulted in 50% attenuation of the effect of DMP 728 on aggregation at 3 hours. Approximately 34% inhibition of the DMP 728-mediated bleeding time was achieved at 1 hour with the 0.2 mg/kg dose, whereas approximately 50% inhibition of the bleeding time was observed for the 1 mg/kg dose at 1 hour.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7489243 TI - Increased elastin-degrading activity and neointimal formation in porcine aortic organ culture. Reduction of both features with a serine proteinase inhibitor. AB - We investigated the association between tissue elastolytic activity and the development of neointimal formation using a previously described porcine aortic organ culture. Neointimal formation is associated with the presence of intact endothelium (nondenuded cultures) but is markedly reduced if endothelial cells are removed (denuded cultures). In nondenuded organ cultures, elastolytic activity assessed by using [3H]elastin increased sixfold at day 3 after initiation of the culture (P < .01), a time earlier than the previously published increase in intimal smooth muscle cells (ISMCs). Elastolytic activity did not increase from day 3 to day 7 despite doubling of ISMCs but did double by day 14 (P < .01) and remained elevated to day 28, correlating with increases in ISMCs. In denuded organ cultures, elastolytic activity was much lower than in nondenuded organ cultures at day 3 (P < .05) but increased fivefold in the presence of nondenuded organ culture conditioned medium (P < .01). Addition of alpha 1 proteinase inhibitor for 14 days caused a 60% decrease in elastolytic activity in nondenuded organ cultures and a 27% reduction in ISMCs compared with untreated controls (P < .05 for both). The elastolytic activity, resolved as lytic bands on an elastin substrate gel, reflected candidate enzymes, one at 76 kD and perhaps a doublet at 43 and 50 kD. Our study suggests that endothelial cells release a soluble agent that enhances elastin-degrading activity in the aorta and may at least partially account for the initiation of neointimal formation. PMID- 7489244 TI - Aldose reductase inhibitor prevents hyperproliferation and hypertrophy of cultured rat vascular smooth muscle cells induced by high glucose. AB - Vascular remodeling is a key process in the pathophysiology of atherosclerosis. Recent evidence suggests that high glucose levels may function as a vascular smooth muscle growth and proliferation-promoting substance. To explore the role of the polyol pathway in this process, we examined the effect of an aldose reductase inhibitor (ARI), epalrestat, on the growth characteristics of cultured rat vascular smooth muscle cells (VSMCs). Epalrestat (10 nmol/L, 1 mumol/L) significantly suppressed the high glucose-induced proliferative effect as measured by [3H]thymidine incorporation by 67% and 82% in cell number, suggesting ARI as an antimitogenic factor. In VSMCs, epalrestat (10 nmol/L, 1 mumol/L) significantly suppressed the high glucose-induced incorporation of [3H]leucine by 45% and 58% with the concomitant reduction of the cell size estimated by flowcytometry. Epalrestat (1 mumol/L) also suppressed high glucose-induced intracellular NADH/NAD+ increase and membrane-bound protein kinase C activation. These results indicate that this ARI possesses an antiproliferative and antihypertrophic action on VSMCs induced by high glucose possibly through protein kinase C suppression. PMID- 7489245 TI - Distribution and correlates of insulin in elderly men. The Honolulu Heart Program. AB - The role of insulin in cardiovascular disease is uncertain, and studies in elderly or minority populations are infrequent. Fasting and 2-hour insulin concentrations and their cross-sectional associations with cardiovascular risk factors were examined in 3562 elderly (aged 71 to 93 years) Japanese American men from the Honolulu Heart Program who were reexamined between 1991 and 1993. Insulin distributions were skewed (mean and median: 16.8 and 12 microU/mL for fasting; 117.2 and 93 microU/mL for 2-hour); fasting but not 2-hour insulin levels declined significantly with age (P < .0001 and P = .54, respectively). Factors most strongly correlated with insulin included measures of obesity, fat distribution, and levels of triglyceride, glucose (r = .38 to r = .50 fasting, r = .21 to r = .27 2-hour), and HDL cholesterol (r = -.41 and r = -.22, respectively). Other correlates included fibrinogen, hematocrit, heart rate, blood pressure, cigarettes per day (all positive), alcohol, physical activity, and forced vital capacity (negative). Associations were also evident across risk factor quintiles. Insulin levels were significantly elevated in men with hypertension and diabetes. In multiple linear regression analyses, log10 fasting insulin was positively and independently associated with body mass index, triglycerides, glucose, fibrinogen, hematocrit, heart rate, diabetes, and hypertension and negatively associated with HDL cholesterol, physical activity, and forced vital capacity. In general, results were similar for log10 2-hour insulin and when subjects who fasted < 12 hours or had diabetes were excluded. Substitution of medication use and blood pressure for hypertension indicated independent associations of medication use but not blood pressure with insulin.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7489246 TI - Relation of a postmortem renal index of hypertension to atherosclerosis in youth. The Pathobiological Determinants of Atherosclerosis in Youth (PDAY) Research Group. AB - In a cooperative multicenter study, Pathobiological Determinants of Atherosclerosis in Youth, of 1164 young men 15 through 34 years of age who died of external causes and were autopsied in forensic laboratories, we measured atherosclerosis of the aorta and the right coronary artery. Using the ratio of intimal thickness to outer diameter of the small renal arteries to predict mean arterial pressure (MAP) during life, we classified the cases as either normotensive (MAP < 110 mm Hg) or hypertensive (MAP > or = 110 mm Hg). By this criterion, the prevalence of hypertension in blacks was 16%; in whites, 12%. Hypertension was associated directly with blood level of glycohemoglobin (an indicator of blood glucose concentration) and with body mass index (BMI) but inversely with thickness of the panniculus adiposus. Among hypertensive compared with normotensive cases, the extent of raised lesions (mainly fibrous plaques) was greater in the aortas of 30- to 34-year-old men and in the right coronary arteries of 25- to 34-year-old men. The prevalence of raised lesions involving 5% or more of the intimal surface was twofold greater in the aortas and right coronary arteries of hypertensive men throughout the 15-to-34-year age span of the study cases. The association of hypertension with raised lesions was not accounted for by adjusting for glycohemoglobin level, BMI, or thickness of the panniculus adiposus. Hypertension is associated with accelerated atherosclerosis in youth. PMID- 7489247 TI - Increased fibrinogen levels in the offspring of hypertensive men. Relation with hyperinsulinemia and the metabolic syndrome. AB - Epidemiological studies have revealed that elevated fibrinogen concentrations are associated with an increased risk of myocardial infarction, stroke, intermittent claudication, and cardiovascular mortality. The manner in which fibrinogen operates in atherogenesis has not yet been elucidated, but genetic control of fibrinogen levels is partially responsible. Fibrinogen frequently acts in concert with hyperlipidemia, diabetes, hypertension, physical inactivity, and age, variables that are influenced by insulin action. Because the offspring of hypertensive men tend to be hyperinsulinemic and insulin resistant from a young age, we hypothesized that their increased fibrinogen levels might reflect decreased insulin action and thus play a role in the metabolic syndrome. We chose 48 adult offspring (mean age, 38.4 years) of 30 fathers who had been treated for hypertension, and the former were matched by age, body mass index, sex, and smoking habits with 37 control subjects. Elevations in fibrinogen concentration (3.63 +/- 0.93 versus 2.87 +/- 0.54 g/L, P < .001) paralleled increases in blood glucose and insulin levels, estimates of insulin resistance, and blood pressure. In the offspring, in contrast to the control group, correlations between fibrinogen and metabolic-syndrome variables (ie, insulin, glucose, and waist and hip circumferences) were found. In stepwise multiple regression analyses, age and smoking habits were entered as variables in both study groups, but postload insulin and high-density lipoprotein cholesterol were entered as variables in the offspring group only. We propose that familial predisposition influences the relationship between insulin concentration and fibrinogen, an effect that may contribute to the clinical importance of the metabolic syndrome. PMID- 7489248 TI - Functional and anatomic evaluation of carotid atherothrombosis. A combined study of indium 111 platelet scintigraphy and B-mode ultrasonography. AB - We examined the relation between in vivo thrombogenicity and the morphology of carotid lesions to clarify the role of platelet deposition in carotid atherothrombosis. We evaluated 60 subjects (120 carotid bifurcations) who had at least one established risk factor for atherosclerosis by using indium 111 platelet scintigraphy and high-resolution B-mode ultrasonography. We evaluated platelet accumulation in the carotid arterial wall by means of a dual-tracer method that used In 111-labeled platelets and technetium 99m-labeled human serum albumin. The tracer accumulation was assessed both visually and semiquantitatively by using the platelet accumulation index, ie, the ratio of radioactivity of the amount of In 111-labeled platelets deposited on the vascular wall to the amount of radioactivity in labeled platelets circulating in the blood pool. The morphology of the carotid lesions was analyzed with B-mode ultrasonography in terms of the presence of ulceration, the maximum percent stenosis, the echogenicity of plaque, and the plaque score, which indicates the severity of systemic atherosclerosis. Platelet accumulation increased with increase in plaque score (P < .01), and the magnitude of platelet accumulation was significantly greater in lesions with ulceration than in those without (P < .05). The platelet accumulation index in vessels with plaque showed a very weak but significant correlation with maximum percent stenosis (r = .28, P < .05) and a stronger correlation with the unilateral plaque score (r = .42, P < .0001). Analysis of the echogenicity of plaque showed that heterogeneous plaque had a high frequency of accumulating platelets. Platelet accumulation was related to the surface characteristics and severity of carotid lesions, especially in the presence of ulceration. PMID- 7489249 TI - Adenovirus-mediated gene transfer to normal and atherosclerotic arteries. A novel approach. AB - Previous studies of gene transfer to blood vessels in vivo have relied on intraluminal, catheter-based methods for delivery of adenoviral and other vectors. In this study, topical application of a replication-deficient adenoviral vector was used as an alternative method of gene transfer to the vessel wall. We administered recombinant adenovirus (1.0 x 1.5 x 10(10) pfu/mL) containing the nuclear targeted bacterial beta-galactosidase gene topically to arteries in normal and atherosclerotic cynomolgus monkeys. Topical administration was achieved by injection of adenoviral suspension within the periarterial sheath. Segments of femoral and carotid arteries were examined histochemically after staining with 5-bromo-4-chloro-3-indolyl-beta-D-galactopyranoside 1 day after treatment with the adenovirus. After topical administration of virus, beta galactosidase activity was observed in approximately 20% of cells in the adventitia in both normal and atherosclerotic arteries. There was no detectable beta-galactosidase activity in cells of the intima or media. Thus, topical application provides an alternative method for gene transfer to blood vessels in vivo. This approach, which does not require interruption of blood flow and does not disrupt the endothelium, may be useful for studies of vascular biology and perhaps gene therapy in both normal and atherosclerotic vessels. PMID- 7489250 TI - Quantitative analysis of repeat adenovirus-mediated gene transfer into injured canine femoral arteries. AB - We quantitatively evaluated the effectiveness of a repeat administration of a recombinant adenoviral vector expressing bacterial Escherichia coli lacZ into the same arterial site of a relatively large animal, the dog. The replication defective adenoviral vector was introduced percutaneously into balloon-injured femoral arteries through a double-balloon catheter. After a single dose of adenoviral vector, up to 90% of surface (73 +/- 16%, n = 7) and smooth muscle cells in multiple layers of the media showed transgene expression as evaluated by 5-bromo-4-chloro-3-indoyl beta-D-galactopyranoside histostaining without extralocal expression, as assessed by polymerase chain reaction. High-level expression (measured as beta-galactosidase activity) peaked 7 days after transfer and was transient, although it was retained for a month. Second does of the same adenovirus to the same arterial site were given 1, 2, 5, or 8 weeks after the first administration. At 1 week the second dose significantly enhanced lacZ expression. At 2, 5, or 8 weeks the second dose reinduced lacZ expression at 25% to 30% of the full expression. lacZ expression was also detected in preimmuned dogs, although the expression levels correlated inversely to the titer of neutralizing antibodies in their serum. These results demonstrate that arterial gene expression can be enhanced by a second administration of the same adenovirus after a short interval and that a repeat dose after a long interval partially but significantly reinduces gene expression despite the presence of an immune response. These data may provide an additional scientific foundation for the use of adenovirus-mediated arterial gene transfer in future clinical practice. PMID- 7489252 TI - Genetic determination of cartilaginous metaplasia in mouse aorta. AB - Calcification frequently occurs in atherosclerotic plaques in humans, but the cellular and genetic factors contributing to this pathological trait are unknown. We previously reported that the arterial calcification among inbred strains is genetically determined, and we now report that cartilaginous metaplasia, associated with the presence of arterial chondrocytes that express type II collagen, may underlie this calcification. Both uncalcified and calcified cartilaginous metaplasia were often colocalized with aortic atheromatous lesions and calcification, and clear genetic differences were observed in the occurrence of aortic cartilaginous metaplasia among inbred strains. Analysis of a genetic cross between strains C57BL/6J (exhibiting aortic cartilaginous metaplasia) and C3H/HeJ (no aortic cartilaginous metaplasia) revealed a recessive inheritance pattern; thus, F1 mice were entirely devoid of cartilaginous metaplasia, in common with the C3H/HeJ parental strain. Analyses of an F2 cross and a set of recombinant inbred strains derived from parental strains C57BL/6J and C3H/HeJ were consistent with a major gene effect exhibiting incomplete penetrance. The occurrence of aortic calcification was correlated with the occurrence of cartilaginous metaplasia in these genetic crosses, suggesting a link between the traits. Finally, we observed widespread calcified cartilaginous metaplasia within spontaneous atherosclerotic lesions in mice targeted for a null mutation in the apoE gene, suggesting that cartilaginous metaplasia is a potential pathway for artery wall calcification associated with the atherosclerotic plaque. PMID- 7489251 TI - Expression and function of a recombinant PDGF B gene in porcine arteries. AB - Platelet-derived growth factor (PDGF) B is a mitogen and chemoattractant for smooth muscle cells in vitro, and expression of a recombinant PDGF B gene in porcine arteries stimulates intimal thickening. To define the mechanisms by which PDGF B gene expression induces intimal thickening in vivo, we examined its effects on smooth muscle cell proliferation and migration, extracellular matrix synthesis, and inflammatory cell infiltration in intimal lesions of pig arteries after direct gene transfer of a recombinant PDGF B gene. PDGF B gene expression was associated with rapid formation of an intima, including 3- to 10-fold increases in intimal thickness and intima-to-media area ratio 4 to 21 days after gene transfer compared with control transfected arteries. Intimal smooth muscle cell proliferation was detected at 2 days, peaked at 7 days (P < .01), and declined by 14 days, although the total number of intimal nuclei progressively increased to 21 days (P < .01). Calculations of expected-to-observed ratios of intimal cells, based on BrdC proliferation indexes, demonstrated that the increases in intimal cell number on days 2 through 7 could not be accounted for by proliferation alone, suggesting that recombinant PDGF BB acts to stimulate cell proliferation and migration of smooth muscle cells into the intima. Extracellular matrix deposition and procollagen synthesis were observed after 7 days (P < .01) and were associated with a decline in cell density in the intima, suggesting that extracellular matrix synthesis may contribute to progressive intimal thickening in response to PDGF B gene expression. There was minimal accumulation of inflammatory cells, including macrophages and CD3(+) lymphocytes, in transfected arteries. These data suggest that PDGF B gene expression promotes intimal expansion by both proliferation and migration of smooth muscle cells followed by synthesis of extracellular matrix and therefore acts through several mechanisms to play a role in the pathogenesis of intimal lesions in vivo. PMID- 7489253 TI - Sodium butyrate inhibits platelet-derived growth factor-induced proliferation of vascular smooth muscle cells. AB - Sodium butyrate (SB), a naturally occurring short-chain fatty acid, was investigated for its therapeutic value as an antiproliferative agent for vascular smooth muscle cells (SMCs). At 5-mmol/L concentration, SB had no significant effect on rat SMC proliferation. However, at the same concentration, SB inhibited platelet-derived growth factor (PDGF)-AA-, -AB-, and -BB-induced proliferation of SMCs. Exposure of SMCs to PDGF-BB resulted in activation of receptor intrinsic tyrosine kinase activity and autophosphorylation of beta-PDGF-receptor (beta PDGFR). The activated beta-PDGFR physically associated and phosphorylated signaling molecules such as ras-GTPase activating protein (GAP) and phospholipase C gamma (PLC gamma). SB, in the absence of PDGF-BB, caused neither beta-PDGFR tyrosine phosphorylation nor phosphorylation and association of GAP and PLC gamma with beta-PDGFR. PDGF-BB-enhanced activation of receptor intrinsic tyrosine kinase activity and autophosphorylation of tyrosine residues of beta-PDGFR were unaffected by SB irrespective of whether SMCs were preincubated with SB before exposure to PDGF-BB plus SB or incubated concomitantly with PDGF-BB plus SB. Likewise, phosphorylation and association of GAP and PLC gamma with PDGF-BB activated beta-PDGFR were unaffected. In addition, SB did not block PDGF-BB stimulated, PLC gamma-mediated production of inositol triphosphate. Similarly, PDGF-BB-induced beta-PDGFR degradation was unaffected when SMCs were exposed to PDGF-BB plus SB, and SB by itself had no influence on beta-PDGFR degradation. Unlike beta-PDGFR kinase activity, mitogen-activated protein kinase (MAP-kinase) activity was stimulated by SB by about 2.7-fold. Exposure of SMCs to PDGF-BB caused an approximately 11.4-fold increase in MAP-kinase activity and this increase in activity was not significantly affected when cells were coincubated with PDGF-BB and SB (10.3-fold). However, pretreatment of SMCs with SB for 30 minutes and subsequent incubation in PDGF-BB plus SB abolished most of the PDGF BB-induced MAP-kinase activity (4.6-fold). Transcription of growth response genes such as c-fos, c-jun, and c-myc were induced by PDGF-BB, and their induction was suppressed, particularly c-myc, by incubating SMCs with PDGF-BB plus SB. Similarly, preincubation of cells with SB for 30 minutes and subsequent incubation in PDGF-BB plus SB diminished PDGF-BB-induced transcription of c-fos, c-jun, and c-myc. However, SB by itself had no significant effect on c-fos, c jun, and c-myc transcription.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 7489254 TI - Human vascular smooth muscle cell-monocyte interactions and metalloproteinase secretion in culture. AB - Degradation of the atherosclerotic plaque extracellular matrix could destabilize the lesion, rendering it more prone to rupture. Both macrophages and vascular smooth muscle cells (SMCs) are potential sources of matrix metalloproteinases (MMPs), secreted enzymes that can digest vascular matrix. We explored interactions between human vascular SMCs and human monocytes that result in the secretion of interstitial collagenase (MMP-1) and stromelysin (MMP-3). Monocytes alone or those treated with SMC-conditioned media did not secrete these metalloproteinases as detectable by Western blot analysis. SMCs increased secretion of both MMP-1 and MMP-3 greater than 20-fold when cocultured with monocytes or when treated with monocyte-conditioned media. Addition of macrophage colony stimulating factor (< or = 1000 U/mL) to cocultures of monocytes and SMCs did not affect metalloproteinase secretion. Recombinant interleukin (IL)-1 receptor antagonist inhibited MMP-1 and MMP-3 induction in SMC cultures treated with monocyte-conditioned media (94% and 96% reduction, respectively), while a neutralizing antibody to tumor necrosis factor-alpha had no significant effect on metalloproteinase secretion. In contrast to the induction by monocyte-conditioned media of MMP-1 and MMP-3 secretion by SMCs, monocyte-conditioned media did not increase secretion of 72-kD gelatinase (MMP-2). Thus, monocytes induce MMP-1 and MMP-3 secretion by vascular SMCs through an IL-1-dependent mechanism. This response of SMCs to a defined macrophage product may contribute to plaque destabilization by mononuclear phagocytes in the lesion. PMID- 7489255 TI - Lysophosphatidylcholine inhibits relaxation of rabbit abdominal aorta mediated by endothelium-derived nitric oxide and endothelium-derived hyperpolarizing factor independent of protein kinase C activation. AB - Hypercholesterolemia is associated with increased oxidized LDL and impaired endothelium-dependent relaxation (EDR). An inhibitory component of oxidized LDL is lysophosphatidylcholine (LPC). To determine the effect and mechanism(s) of action of LPC on EDR mediated by endothelium-derived nitric oxide (EDNO) and endothelium-derived hyperpolarizing factor (EDHF), rabbit abdominal aortic rings were suspended for measurement of isometric tension and studied under three conditions: control; with 25 mmol/L K+ buffer to isolate relaxation mediated by EDNO; and in rings treated with N omega-nitro-L-arginine methyl ester (L-NAME, 30 mumol/L) to isolate relaxation mediated by EDHF. Incubation with LPC (10 and 30 mumol/L) for 30 minutes inhibited EDR in a concentration-dependent manner. LPC (30 mumol/L) significantly inhibited maximal relaxation to acetylcholine in control, 25 mmol/L K(+)-, and L-NAME-treated rings (77.1 +/- 7.8%, 42.1 +/- 8.9%, and 3.4 +/- 7.7%) compared with untreated rings (99.0 +/- 0.9%, 90.9 +/- 2.2%, and 54.7 +/- 4.7%, P < .05). Inhibition of relaxation was specific to endothelium dependent responses in that relaxation to direct smooth muscle vasodilators (papaverine, 8-bromo-cGMP, and sodium nitroprusside) were unaltered by LPC. The inhibition by LPC (30 mumol/L) was not due to cytotoxicity, because EDR returned to normal levels after repeated washing with physiological salt solution containing 0.1% albumin. Co-incubation with protein kinase C inhibitors, staurosporine (20 nmol/L) or calphostin C (1 mumol/L), had no effect on the EDR inhibition by LPC (30 mumol/L). Furthermore, LPC continued to inhibit EDR in rings in which protein kinase C was down-regulated by incubation for 18 hours with 1 mumol/L phorbol 12-myristate 13-acetate (PMA).(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7489256 TI - Activation of protein kinase C increases adenosine production in the hypoxic canine coronary artery through the extracellular pathway. AB - Both ischemia and hypoxia increase adenosine production in the heart. This study tested whether hypoxia increases adenosine production in the coronary artery via ecto-5'-nucleotidase and the role of protein kinase C in this condition. Canine left circumflex coronary artery was rapidly removed and incubated in 10 mL Krebs Henseleit solution for 30 minutes. The Krebs-Henseleit solution contained 5' iodotubercidin and 2'-deoxycoformycin, which inhibit adenosine kinase and adenosine deaminase, respectively. Adenosine production was measured in intact coronary arteries under normoxic conditions (16.2 +/- 1.2 pmol/mg protein). Adenosine production was reduced by 27% after removal of endothelium. Ecto-5' nucleotidase activity of coronary arteries with and without endothelium was 51 +/ 6 and 41 +/- 4 nmol/mg protein per minute under normoxic conditions. Hypoxia increased adenosine production to 27.0 +/- 2.3 and 20.0 +/- 0.8 pmol/mg protein with and without endothelium. Hypoxia also increased ecto-5'-nucleotidase activity of coronary arteries with and without endothelium (74 +/- 8 and 53 +/- 5 nmol/mg protein per minute; P < .05). Increases in adenosine production under hypoxic conditions were blunted by both an inhibitor of ecto-5'-nucleotidase and inhibitors of protein kinase C. Activation of ecto-5'-nucleotidase was blunted by an inhibitor of protein kinase C. These results indicate that hypoxia increased extracellular adenosine production and activated ecto-5'-nucleotidase via activation of protein kinase C in coronary arterial smooth muscle and endothelial cells. Increased adenosine production in coronary arteries during hypoxia may contribute to coronary vasodilation and cardioprotection against ischemic injury. PMID- 7489257 TI - Oxidized LDL-induced microvascular dysfunction. Dependence on oxidation procedure. AB - Human LDLs oxidized with Cu2+ are known to promote leukocyte-endothelial cell adhesion (LECA) and albumin leakage in postcapillary venules. The objective of this study was to compare the ability of LDL oxidized with Cu2+ (Cu-LDL), phospholipase A2 plus lipoxygenase (PLA2-LDL), horseradish peroxidase plus H2O2 (HRP-LDL), or -OCl (-OCl-LDL) to promote (1)neutrophil-endothelial cell adhesion (NECA) in vitro and (2)LECA and albumin leakage in rat mesenteric venules. In vitro adhesion assays revealed that only Cu-LDL elicited a dose-dependent NECA response, whereas PLA2-LDL but not normal (N-LDL), HRP-LDL, or -OCl-LDL increased NECA at the highest concentration studied (670 micrograms/mL). The magnitude of the NECA responses elicited by the different forms of oxidized LDL was related to the degree of lipid peroxidation but unrelated to the level of protein oxidation. Local intra-arterial infusion of Cu-LDL, PLA2-LDL, or -OCl-LDL but not N-LDL elicited significant increases in leukocyte adherence and emigration, mast cell degranulation, and albumin leakage in rat mesenteric venules. The LECA induced by all forms of oxidized LDL was not accompanied by significant alterations of venular shear rate. PMID- 7489258 TI - Introduction. House dust mite allergy. PMID- 7489259 TI - The biology of allergenic domestic mites. An update. PMID- 7489260 TI - Dust mite allergens. PMID- 7489262 TI - Atopic dermatitis and delayed hypersensitivity to dust mites. PMID- 7489261 TI - Specific immunotherapy in house dust mite allergy. PMID- 7489264 TI - Effects of maxillary anterior supernumerary impacted teeth on diastema. PMID- 7489263 TI - "I'd really like to be an oral pathologist, but...". PMID- 7489266 TI - Steepness of the articular eminence in the temporomandibular joint. Tomographic comparison between asymptomatic volunteers with normal disk position and patients with disk displacement. AB - OBJECTIVE: A steep articular eminence has been proposed as one etiologic factor for the development of TMJ disk displacement. The purpose of this study was to determine the steepness of the articular eminence in asymptomatic volunteers with superior disk position and to compare it with the articular eminence in patients with internal derangement. STUDY DESIGN: We investigated 34 asymptomatic volunteers and 71 patients. The steepness of the articular eminence was determined on sagittally corrected tomograms, and disk position was diagnosed by dual space arthrotomography. RESULTS: The results showed that the articular eminence was on the average steeper in the asymptomatic volunteers in the lateral (64.9 degrees), central (64.4 degrees), and medial (65.4 degrees) sections of the joint than in the patients (56.1 degrees, 60.2 degrees, 58.9 degrees, respectively). When the comparison of the steepness of the eminence was based on the presence or absence of osseous changes in the form of remodeling or osteoarthrosis, no difference was found between the asymptomatic volunteers and the patients with disk displacement but without such changes. The largest difference in the steepness of the eminence was found between the persons with osseous changes and those without. CONCLUSION: These findings contradict earlier studies that have indicated a steep articular eminence to be one etiologic factor for the development of disk displacement. The results of this study showed that the steepness of the articular eminence is decreased in the patients as a result of remodeling or degenerative changes of the bone that are a result of the internal derangement. PMID- 7489268 TI - The Christensen prosthesis. A retrospective clinical study. AB - The objective of this study was to determine if the Christensen temporomandibular joint prosthesis system in an effective alternative in treating patients with severe temporomandibular joint disorders. A total of 69 patients who were not responsive to either nonsurgical or prior surgical treatments were placed into one of three treatment groups depending on the following diagnoses: (1) placement of a glenoid fossa-eminence prosthesis with meniscus retention (22 patients, 40 joints); (2) placement of a glenoid fossa-eminence prosthesis without retention of the meniscus (26 patients, 49 joints); (3) total joint replacement (21 patients, 34 joints). Patients were evaluated immediately before surgery and at regular intervals after surgery for an average of 3.1 years. Success was measured as a significant improvement of function and decrease in pain as measured on a visual analogue scale, as well as improved incisor opening as measured with a Therabite Scale (Lorenz Surgical, Jacksonville, Fla.). Comparison of mean and average pre- and postsurgical values for all groups and criteria showed significant improvement. Results of this study indicate that the Christensen temporomandibular joint prosthesis system may offer a viable method for the treatment of severe temporomandibular joint disease with a high degree of success. PMID- 7489267 TI - Total reconstruction of the temporomandibular joint with cryogenically preserved allograft mandibular condyle, meniscus, and fossa in the adult goat. AB - OBJECTIVE: The purpose of this study was to determine the feasibility of total temporomandibular joint reconstruction with the use of cryogenically preserved allograft mandibular condyle, disk, and fossa in the adult goat. STUDY DESIGN: This study consisted of an experimental group of 10 adult goats that underwent total resection of their temporomandibular joints. These animals were immediately reconstructed with cryogenically preserved mandibular condyles, disks, and fossa. These allograft were harvested from 10 donor animals matched to the recipient animal with respect to age, sex, and head dimensions. These grafts were harvested and stored at -185 degrees C for several months before implantation. At 2 years after surgery, the animals were killed. They were evaluated, clinically, radiographically, and histologically. RESULTS: Fifty percent of the eight surviving animals had a normal clinically functional joint with an intact superior joint space and viable articular surfaces. CONCLUSION: The authors conclude that this temporomandibular joint reconstruction technique is feasible in the adult goat model and is deserving of further research. PMID- 7489265 TI - Short-term treatment outcome study for the management of temporomandibular joint closed lock. A comparison of arthrocentesis to nonsurgical therapy and arthroscopic lysis and lavage. AB - OBJECTIVE: The aim of this study is to compare the clinical short-term results and efficacy of arthrocentesis with those of nonsurgical treatments and arthroscopic surgery for the management of temporomandibular joint closed lock. STUDY DESIGN: The three groups consisted of 63 consecutive patients treated nonsurgically, 20 patients treated with arthrocentesis, and 25 patients treated with arthroscopic surgery. All patients rated their pain level on a visual analogue scale and completed a pain, jaw dysfunction, and activity-limiting questionnaire before and 6 months after the procedure. Each patient's jaw opening was also scaled. The success rate of each procedure was calculated according to our success criteria. The clinical efficacy of each procedure was evaluated in successfully treated subgroups in which the between-group and within-group differences of the obtained data were statistically tested. RESULTS: The success rate was 55.6% in the nonsurgically treated group, 70% in the arthrocentesis group, and 91% in the arthroscopy group. With respect to efficacy, all posttreatment scores showed significant improvements in within-group differences, but no between-group differences were found between the arthrocentesis group and the other two groups. CONCLUSION: Arthrocentesis was considered as an intervening treatment modality between nonsurgical treatment and arthroscopic surgery on the basis of its short-term outcome. Although the treatment efficacy was comparable with arthroscopic surgery, this procedure was thought to be indicated for the patients with acute temporomandibular joint closed lock who were refractory to medication and mandibular manipulation rather than the alternative of arthroscopic surgery. PMID- 7489269 TI - The effect of polylactic acid granules on the incidence of alveolar osteitis after mandibular third molar surgery. A prospective randomized study. AB - OBJECTIVE: To evaluate the effect of polylactic acid granules placed in third molar sockets on the incidence of alveolar osteitis. STUDY DESIGN: A prospective randomized study with 161 patients (322 bilaterally impacted mandibular third molars). RESULTS: The sockets in which the polylactic acid granules were placed had a significantly higher rate of alveolar osteitis (23.6%) than did the control sockets (13.58%). In all subgroups analyzed, the incidence of alveolar osteitis was higher in the experimental (polylactic acid) group. CONCLUSION: On the basis of the findings of this study, polylactic acid granules do not reduce the incidence of alveolar osteitis and may actually contribute to a higher incidence of alveolar osteitis. PMID- 7489270 TI - Relationship between salivary flow rates and Candida albicans counts. AB - Seventy-one persons (48 women, 23 men; mean age, 51.76 years) were evaluated for salivary flow rates and Candida albicans counts. Each person was seen on three different occasions. Samples of unstimulated whole, chewing-stimulated whole, acid-stimulated parotid, and candy-stimulated parotid saliva were collected under standardized conditions. An oral rinse was also obtained and evaluated for Candida albicans counts. Unstimulated and chewing-stimulated whole flow rates were negatively and significantly (p < 0.001) related to the Candida counts. Unstimulated whole saliva significantly (p < 0.05) differed in persons with Candida counts of 0 versus <500 versus < or = 500. Chewing-stimulated saliva was significantly (p < 0.05) different in persons with 0 counts compared with those with a > or = 500 count. Differences in stimulated parotid flow rates were not significant among different levels of Candida counts. The results of this study reveal that whole saliva is a better predictor than parotid saliva in identification of persons with high Candida albicans counts. PMID- 7489271 TI - An unusual case of the Melkersson-Rosenthal syndrome. A case report. AB - A case of Melkersson-Rosenthal syndrome in a woman with a history of trauma and infection is presented. In addition to the classical triad of signs, the patient also exhibited several other features of rare occurrence associated with Melkersson-Rosenthal syndrome. Clinical diagnosis was confirmed by biopsy, which revealed the characteristic features of Melkersson-Rosenthal syndrome. PMID- 7489272 TI - Case of bilateral non-Hodgkin's lymphoma after dental extractions in a patient with the human immunodeficiency virus. AB - It has long been known that immunocompromised patients have an increased risk of getting malignant neoplasms, with lymphomas predominating. This increase in lymphomas is especially notable in the population infected with the human immunodeficiency virus (HIV). Before the outbreak of the human immunodeficiency virus these neoplasms rarely occurred in the maxillofacial region; they are now being seen with a greater frequency. This report details a case of bilateral non Hodgkin's lymphoma that was initially misdiagnosed as an odontogenic infection. PMID- 7489273 TI - Myofibromas of the oral cavity. AB - Solitary infantile myofibromatosis or myofibroma of the oral cavity is an uncommon condition with only 32 reported cases in the English-language literature. This article presents four additional cases of these solitary myofibroblastic lesions. In addition, the clinical and histologic features of this uncommon spindle cell neoplasm have been reviewed. The similarity in both the clinical and histopathologic features of the "adult" and "infantile" lesions support the proposal that myofibroma is a more accurate and acceptable term for these solitary myofibroblastic lesions of the oral cavity. PMID- 7489274 TI - Myofibroma of the mandible. Clinicopathologic study and review of the literature. AB - A case of mandibular myofibroma in a 2-month-old boy is presented. Including this case, 24 pediatric and 11 adult patients with maxillofacial myofibroma have been reported since 1981. Of the 24 pediatric patients, 15 (62.5%) had lesions affecting the mandible. The adult cases had no mandibular involvement. Histologic evaluation of the tissue specimen revealed an interlacing pattern of spindle shaped cells with long oval nuclei. Tissue immunohistochemical staining found it to be reactive for antibodies directed against vimentin and alpha-smooth muscle actin, but not desmin, S-100 protein, neuron-specific enolase, or myoglobin. Electron microscopy examination revealed the following cells: myofibroblast-like cells, fibroblast-like cells, and intermediate cells that were similar to the fibroblast-like cells except for the presence of a few microfilaments. Myoblast like cells were not seen. PMID- 7489275 TI - Primary leiomyosarcoma of the maxilla with regional lymph node metastasis. Report of a case and review of the literature. AB - A rare case of oral leiomyosarcoma diagnosed with the aid of immunohistochemical and electron microscopic examinations together with a review of the literature are reported. The patient was a 70-year-old Japanese man. The primary tumor involved the maxillary gingiva and bone and metastasized to the cervical lymph nodes. On histologic examination the tumor showed invasive growth into the maxillary bone. It was composed of interlacing fascicles of spindle-shaped cells with eosinophilic cytoplasm and elongated, blunt-ended nuclei. The tumor formed extensive metastatic foci in the cervical lymph nodes. On immunohistochemical examination most of the tumor cells were positive for desmin, smooth muscle specific actin, and myosin. The ultrastructural characteristics of the tumor cells were abundant microfilaments, pinocytotic vesicles, and basement membrane formation. The findings were indicative of a tumor demonstrating myogenic differentiation. A review of the literature during the past 50 years disclosed a total of 60 oral leiomyosarcomas, including our case. PMID- 7489276 TI - Prevalence of human papillomavirus 16 and nucleolar organizer region counts in oral exfoliated cells from normal and malignant epithelia. AB - This study was designed to identify the prevalence of human papillomavirus 16 (HPV 16) in oral exfoliated cells from 26 patients with oral cancer and matched healthy volunteers with the use of polymerase chain reaction. In addition, the value of a silver staining technique for nucleolar organizer regions (AgNORs) was also investigated. HPV 16 was detected in 30.8% of the cancer lesions, 26.9% of the unaffected sites, and 15.4% of samples from normal mucosa. AgNOR counts on the same cases were analyzed. Although AgNOR counts are useful in distinguishing between normal and malignant oral exfoliated cells, they provided no additional prognostic information for oral cancer. However, when AgNOR counts were compared with HPV 16-positive and HPV 16-negative counts in cancer lesions, AgNOR counts were higher in HPV-positive lesions. These findings suggest that HPV 16 may play a role in tumor cell proliferation, but it is unlikely to play a significant role alone in the cause of oral cancer. Therefore evidence of HPV 16 infection in oral malignant neoplasms should be cautiously interpreted. PMID- 7489277 TI - Human pulp response after partial pulpotomy with two calcium hydroxide products. AB - Twenty human permanent teeth were used to study the pulpal response of two calcium hydroxide products, Dycal and Pulpdent Multi-Cal, after partial pulpotomy. Teeth were extracted at 4 months, fixed, and prepared for histologic examination. All 10 teeth treated with Dycal showed complete soft tissue healing and bridge formation. No stained bacteria were seen throughout the serial sections. One tooth treated with Dycal showed acceptable histologic results, dentin deposition in the root canal. Six cases dressed with Pulpdent Multi-Cal showed acceptable histologic results, whereas four teeth showed severe inflammation or necrosis associated with bacterial penetration into the pulp tissue. Clinically, one tooth treated with Pulpdent Multi-Cal showed pulpal pain and was extracted at 90 days. Our data support the thesis that human permanent pulps will promote tissue healing and dentin bridge formation as long as bacterial microleakage is excluded. PMID- 7489278 TI - Effect of thickness on the sealing ability of some root canal sealers. AB - A fluid transport model study was used to evaluate the sealing ability of five root canal sealers, AH 26, Sealapex, Ketac Endo, Roth, and Kerr EWT, and a dental bonding agent at thicknesses of 0.05 mm (thin layer) and 0.3 mm (thick layer) with 270 standard human root sections obturated with sealer combined with standard gutta-percha cylinders. AH26 and Sealapex sealed more tightly in thick layers, whereas Ketac Endo, Johnson and Johnson Bonding Agent, Roth, and Kerr EWT sealed more tightly in thin layers. In thin layers J&J Bonding sealed more tightly than any other sealer tested. In thick layers AH 26 and Sealapex sealed more tightly than Ketac Endo and J&J Bonding Agent, and these sealed more tightly than Roth and Kerr EWT. These findings indicate that the thickness of the sealer layer significantly influences the sealing of a root canal filling and that the influence of thickness varies depending on the type of sealer. PMID- 7489279 TI - Absolute measures of image quality for the Sens-A-Ray direct digital intraoral radiography system. AB - To study the noise characteristics of the Sens-A-Ray (Regam Medical Systems AB, Sundsvall, Sweden) system, 20 radiographs were obtained at each of three different exposure levels at 70 and 90 kVp with a homogeneous x-ray field. Exposures were measured with an ionization chamber. Noise power spectra were calculated over three areas within each radiograph, and ensemble averages were subsequently found from 60 data files at each exposure level. Noise equivalent quanta were calculated with the noise power spectra and modulation transfer function data from previous studies. Finally, the detective quantum efficiency was calculated by dividing the noise equivalent quanta by the estimated incident photon fluence at the different exposures. The system has a maximum detective quantum efficiency of approximately 0.030 at 70 kVp and 0.025 at 90 kVp. A broad maximum exists at approximately 2 cycles/mm, indicating that the signal-to-noise ratio is most favorable at this spatial frequency. PMID- 7489280 TI - Radiographic evaluation of implant-obscured bone. Comparison of digitally subtracted tomographic and periapical techniques. AB - This study evaluated the diagnostic accuracy of digitized and substracted periapical and tomographic film images in the assessment of facial and lingual bone loss at implant-obscured sites. An edentulous dried human mandible was prepared with implants in the incisor, premolar, and molar regions. Successive bur sizes from quarter round to no. 6 were used to create circumferential infrabony periodontal defects in the alveolar crest adjacent to the implants. Periapical radiographs were made with the use of custom stents. Linear tomograms imaged sites in a cross-sectional orientation. An equal number of radiographs with and without lesions were digitized and subtracted from baseline images. Radiographic anatomy on the mesial and distal aspects of each implant was electronically masked before the evaluation by a series of eight observers. Receiver operating characteristics analysis and analysis of variance were used to evaluate accuracy of interpretation of the presence or absence of crestal defects. Observer accuracy in detecting facial or lingual bone defects was significantly greater with subtracted tomographic images than with substracted periapical images (p < 0.001). PMID- 7489281 TI - Reliability of a paralleling instrument for dental radiographs. AB - The reliability of a paralleling instrument for dental radiographs was retrospectively evaluated in the course of a 6-month clinical study. The angular variation between successive exposures was precisely quantified. Ninety-one percent of the angular variations were below a 1.4 degree threshold, but some paired images were found unsuitable for quantitative analysis. A mathematic procedure to assess the global angular projection error of any similar coupled x ray cone-film holder device is proposed. This mathematic procedure should be routinely used to check superimposability of serial radiographs before density quantification or subtraction. PMID- 7489282 TI - A survey of the radiographic practices of general dentists for edentulous patients. AB - OBJECTIVE: The objective of the study was to evaluate the radiographic practice of general dentists for edentulous patients. STUDY DESIGN: A questionnaire was sent to a sample of graduates from the two Ontario dental schools. RESULTS: The adjusted response rate for the survey was 80%. Fifty-three percent of the respondents used screening radiography for new patients. The overwhelming choice for these dentists was the panoramic film. There was a significant positive association between ownership of a panoramic unit and use of this radiograph. Dentists who do not own a panoramic unit rarely chose to refer their patients for this radiograph either routinely or selectively. When dentists, rather than the auxiliaries, made the greater proportion of the radiographs, they tended to use selective radiography. Most dentists chose selective radiography for the recall patient. CONCLUSIONS: Almost half the dentists did not follow the Food and Drug Administration/American Dental Association guideline of screening radiography for the new edentulous patient. Of those dentists who did follow the guideline the vast majority both owned panoramic units and chose to use a panoramic survey. Most dentists followed the guideline for selective radiography for the recall patient. PMID- 7489283 TI - Recurrent lower gingival squamous cell carcinoma spreading along the pathway of the inferior alveolar nerve. AB - Computed tomography images showed recurrent lower gingival squamous cell carcinoma spreading from the mandibular foramen to the middle cranial fossa. This extensive tumor spread followed exactly the same anatomic pathway as the inferior alveolar nerve. Histopathologic examination revealed squamous cell carcinoma. Tumor spread was thought to be caused by perineural invasion. This case demonstrates computed tomography features of perineural tumor spread along the inferior alveolar nerve. PMID- 7489284 TI - Magnetic resonance imaging techniques in the pelvis. AB - Basic and more recent advances in imaging the pelvis are reviewed in this article. Commercially available innovations such as phased array multicoils in fast spin echo imaging can be used to improve resolution for MR imaging of the pelvis. Chemical shift imaging and vascular imaging in the pelvis also are reviewed, and potential roles for gadolinium-enhanced imaging and gastrointestinal contrast agents are discussed. PMID- 7489287 TI - Magnetic resonance imaging of the ovary. AB - The authors present the newer MR imaging techniques and how to use them to diagnose the common benign and malignant causes of adnexal masses. Specific features on MR imaging for adnexal lesions that are stressed include morphologic features, specific chemical shift, magnetic susceptibility, and enhancement pattern. PMID- 7489285 TI - Magnetic resonance imaging of the uterus. AB - MR imaging has emerged as the most accurate imaging modality for the evaluation of many conditions affecting the uterus. This technique can both define the normal internal anatomy of the uterus and monitor the physiologic responses of this organ to various endogenous and exogenous stimuli. MR imaging provides unique diagnostic information that may guide therapeutic management of developmental anomalies, adenomyosis, and endometrial carcinoma. PMID- 7489286 TI - Magnetic resonance imaging applications in uterine cervical cancer. AB - We have discussed the importance of staging in making decisions about the type and extent of surgery and in determining prognosis, and at the same time have noted the remarkably poor accuracy of clinical staging. We have detailed the present and potential accuracy of MR imaging for assessing size and extent of tumor and its advantages compared to clinical and other (primarily CT) staging. The question that naturally arises is that if MR imaging is so good, why isn't it used more often? In a critical review of the use of CT for staging, Moore et al noted that "Ultimately, a diagnostic test can be considered useful only if it provides information leading to a change in therapy with patient benefit." In a retrospective review of 246 patients divided into those who did and those who did not undergo pretreatment CT, they found that only 8 patients had improved survival from treatment modifications based on CT, while in the same group 8 patients underwent additional surgical procedures because of CT findings that proved to be erroneous. They concluded that "Considering the high cost and limited benefit, CT for cervical cancer staging is not recommended." Although this sort of critical outcome analysis has not yet been done for MR imaging, it is evident from our discussion that MR imaging has much to offer that CT does not, and that MR imaging can indeed provide information leading to a change in therapy. Unlike CT, MR imaging can measure accurately the size of the tumor, determine whether or not it is confined within the cervix, and determine extension to the vagina, parametrium, or myometrium. MR imaging even has advantages in assessing lymph node involvement and is particularly recommended when clinical tumor diameter approaches 3 cm, in which case the incidence of lymph node metastasis approaches 50%. With rapid improvements in techniques and hardware, especially dedicated coils, fast acquisition pulse sequences, and dynamic enhancement methods, it can be expected that MR imaging will become even more accurate in identifying and staging disease. Given this, the question still remains as to why MR imaging is not used more in the work-up of patients with cervical cancer. In part this may be caused by na lack of awareness by clinicians of the advantages of MR imaging in this application. In part this may be caused by limited availability of MR imaging systems, although by now most midsized and probably all large hospitals have at least one MR imaging unit.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 7489288 TI - Magnetic resonance imaging of benign adnexal conditions. AB - Magnetic resonance imaging can be used to demonstrate a variety of benign adnexal diseases. However, the real utility and efficacy of MR imaging in benign pelvic disease has not been investigated thoroughly. This is the challenge for future research. Presently, particularly in light of changes in the health care system, it is incumbent upon radiologists, gynecologists, and other clinicians to determine what additional information MR imaging will add to the diagnostic evaluation already completed before performing MR imaging. One must ask several questions. Will MR imaging provide additional information? Will that information affect care and treatment of the patient? Will the treatment be beneficial to the care of the patient? Will the patient agree to the management? Many times these questions are not asked. And if asked, the answers are unknown or under investigation. However, using one's best clinical judgment and armed with the knowledge at hand, if answers to the above questions are yes, then MR imaging may prove useful. PMID- 7489290 TI - Pregnancy and the safety of magnetic resonance imaging. AB - Many data exist regarding the safety of the static and time-varying magnetic and electromagnetic fields used in MR environments for the developing fetus. Many of these data, however, are confusing and nebulous. The literature in this area is reviewed, and the issues related to MR imaging of the pregnant patient and MR and the pregnant health care practitioner are discussed. Practice guidelines also are suggested. PMID- 7489289 TI - Magnetic resonance imaging in pregnancy. AB - MR imaging can be important to demonstrate nonpelvic, pelvic, and fetal abnormalities in the pregnant patient. New, faster techniques expand diagnostic capabilities. Although no convincing evidence for fetal injury has been demonstrated, a rational approach to MR imaging in pregnancy is proposed in this article. PMID- 7489291 TI - MR imaging of the menisci. Fundamentals of anatomy and pathology. AB - This article discusses the anatomy of the menisci and various signal abnormalities. Morphologic changes indicative of degenerative disease, degenerative tear, intra- and para-meniscal cysts, and traumatic tear are also covered. PMID- 7489292 TI - Magnetic resonance imaging of the meniscus. Advanced concepts. AB - Magnetic resonance imaging plays a valuable role in the evaluation of the full range of meniscal pathology including the primary diagnosis of meniscal tear, the detection of recurrent tear after resection or repair, and the demonstration of associated injuries and complications. The accuracy of MR imaging in meniscal diagnosis is dependent on the appropriate choice of MR imaging pulse sequences, imaging parameters, and diagnostic criteria. The importance of small details such as protocol parameters and image photography should not be underestimated. The role of new techniques including fast imaging sequences, three-dimensional imaging, and MR arthrography in improving meniscal diagnostic performance is under investigation and appears promising. PMID- 7489293 TI - MR imaging of anterior cruciate ligament injuries. AB - MR imaging has become the imaging modality of choice to evaluate the integrity of the anterior cruciate ligament (ACL). This article discusses the normal anatomy of the ACL and the clinical diagnosis of ACL disruption. The MR imaging appearance of chronic and acute ACL injuries, and the relative value of primary and secondary signs in injury diagnosis are reviewed. The clinical value of MR imaging in the evaluation of the ACL-deficient knee is also discussed. PMID- 7489294 TI - Medial and lateral supporting structures of the knee. Normal MR imaging anatomy and pathologic findings. AB - The supporting structures of the knee can be divided into several layers: fascial boundaries, the collateral ligaments and tendons, and the joint capsule. MR imaging's ability to demonstrate these findings has helped to validate this anatomic concept. An appreciation for the organization of the medial and lateral compartments facilitates MR imaging interpretation. PMID- 7489295 TI - Magnetic resonance imaging of the extensor mechanism. AB - Trauma involving the extensor mechanism of the knee is a fairly common clinical problem, and can be addressed easily with a basic understanding of the anatomy of the involved structures. Injury to the muscles and tendons of this group is categorized broadly into complete versus partial tears based on the respective absence or presence of intact fiber bundles. Acute injuries are associated with edema, hemorrhage, and fluid collections; chronic injuries often demonstrate redundancy, atrophy, and retraction. Fracture of the patella can be related to direct or indirect trauma, and is functionally equivalent to a complete tear of the musculotendinous elements. Tendinitis is a more chronic condition seen with repetitive trauma, and usually affects the proximal aspect of the patellar tendon. Swelling and abnormal intrasubstance signal are commonly associated with this entity. PMID- 7489296 TI - Magnetic resonance imaging of bone marrow and osteochondral injury. AB - Posttraumatic occult subcortical fractures represent a heterogeneous group of lesions, with different subgroups associated with a high incidence of osteochondral sequelae. The role of MR imaging must be to determine the significance of the different subgroups of occult fractures in the pathogenesis of posttraumatic osteoarthritis. The ultimate goal is to minimize the eventual effects of impactive forces on the osteochondral complex at the time of detection. Modifications in immediate and rehabilitative management of such patients will ensure maintenance of the best possible chondral integrity. PMID- 7489297 TI - MR imaging of the patellofemoral compartment. AB - This article reviews the applications of MR imaging of the patellofemoral compartment. Axial plane images are the most informative for abnormalities of this compartment. The role of MR imaging in the evaluation of the medial synovial plica and in the detection of chondromalacia is discussed. MR imaging can reliably detect and delineate the complex of injuries associated with patellar dislocations and valgus hyperextension. PMID- 7489298 TI - Kinematic magnetic resonance imaging of the knee. AB - The emergence of kinematic MR imaging has added a new dimension to imaging of the knee. Ultrafast scan techniques can be used to detect patellofemoral malalignment and tracking abnormalities. Sensitivity can be increased by using a quadriceps loading device. PMID- 7489299 TI - Magnetic resonance imaging in synovial disorders and arthropathy of the knee. AB - The ability of MR imaging to demonstrate articular cartilage, the synovium, and the adjacent bone greatly assists in the diagnosis and treatment of synovial disorders and a variety of arthropathies of the knee. The use of specific pulse sequences and intravenous contrast material can improve the performance of MR imaging of the knee in patients with synovitis, arthritis, or synovial-based neoplasms. PMID- 7489301 TI - Cost effectiveness of magnetic resonance imaging of the knee. AB - When compared to diagnostic arthroscopy in patients with knee complaints consistent with internal derangement, MR imaging emerges as a cost effective diagnostic supplement to clinical examination. This article concludes that the results of MR imaging can be used as a guide for the management of knee pain. PMID- 7489302 TI - Contemporary magnetic resonance imaging of the knee. The orthopedic surgeon's perspective. AB - This article discusses the current applications of MR imaging in the practice of orthopedic surgery and sports medicine. When is MR imaging helpful to the treating surgeon, and when is it not? In what specific areas will future improvements in technical acquisition and interpretation be helpful in determining appropriate treatment? This article explores these questions. PMID- 7489300 TI - Magnetic resonance imaging of bone marrow disorders of the knee. AB - Bone marrow abnormalities around the knee are encountered frequently when performing MR imaging. Some of these so-called abnormalities may represent normal variations of the bone marrow contents and have no clinical significance. On other occasions, MR imaging may be the only technique that demonstrates a bone marrow lesion. Edema, osteonecrosis, infection, and tumor are entities frequently detected and assessed with MR imaging. We now, perhaps for the first time, are able to visualize and assess directly the bone marrow and its disorders. Knowledge of normal anatomy, variants, and pathologic signs coupled with meticulous imaging techniques are of paramount importance for accurate interpretation of such MR images. PMID- 7489303 TI - Magnetic resonance imaging of the knee. A senior musculoskeletal radiologist's perspective. AB - After a brief review of the historical development of knee arthrography and MR imaging of the knee, the author concludes that MR imaging has become widely accepted as the imaging method of choice by both radiologists and nonradiologists. The author examines the current relative shortcomings and future developments of MR imaging of the knee. PMID- 7489305 TI - Imaging techniques for breast MR imaging. AB - Specialized rf coils and pulse sequence techniques for contrast-enhanced breast imaging have been a focus of recent attention in MR imaging research. The imaging strategies for maximizing sensitivity and specificity involve trade-offs between spatial and temporal resolution. This article discusses these issues and others, such as considerations for coil design, fat suppression, patient positioning, motion artifact reduction, and methods for quantitative measurement of dynamic contrast enhancement. PMID- 7489304 TI - MR spectroscopy and imaging of breast cancer. A brief history. AB - The application of nuclear magnetic resonance (NMR) techniques to the diagnosis of breast cancer has come a long way in a relatively brief period of time. Early investigators focused on diagnosis by in vitro NMR spectroscopy. More recently, in vivo NMR spectroscopy, contrast-enhanced MR imaging, and MR imaging-controlled biopsy have been investigated. The author predicts that MR imaging of breast cancer quickly will make the transition from experimental method to useful clinical technique. PMID- 7489306 TI - Sensitivity of contrast-enhanced MR imaging of the breast. AB - High sensitivity is a major advantage of contrast-enhanced MR imaging when it is applied as an additional tool in difficult cases. Several factors, such as technique, guidelines, patient preselection, and varying levels of verification influence sensitivity and specificity of contrast-enhanced MR imaging. This article explores these issues, and reviews data based on the authors' own experience and on the published literature. PMID- 7489307 TI - False-positive results in dynamic MR mammography. Causes, frequency, and methods to avoid. AB - Numerous studies have shown that MR mammography (MRM) yields the highest sensitivity in the diagnosis of early and small breast cancers. The multifocality of breast cancers can be recognized only with MRM. This article evaluates the false-positive results of dynamic MRM. The causes of false diagnoses are also examined and ways to avoid them are suggested. PMID- 7489308 TI - The specificity of contrast-enhanced breast MR imaging. AB - The sensitivity of contrast-enhanced breast MR imaging for malignancy is close to 100%, an improvement over traditional methods of cancer detection. The reported specificity of breast MR imaging, however, ranges from approximately 37% to 97%. This range of values is the result of differences in technique and diagnostic criteria. The variation in biologic activity and vascularity of benign and malignant lesions, as well as normal tissue, also complicates MR imaging diagnosis. A review of breast MR imaging research with an emphasis on specificity issues is also presented. PMID- 7489309 TI - Staging of breast cancer with MR imaging. AB - The vastly improved sensitivity of new MR methods can be used to define disease within the breast that cannot be seen with conventional breast imaging methods. Breast MR imaging is expected to have a significant role for the staging of breast cancer in breast conservation candidates. Ultimately, breast MR will be integrated with interstitial laser photocoagulation as a treatment method for breast cancer. PMID- 7489310 TI - MR guided biopsy of the breast. AB - This article examines technical and clinical issues related to MR guided biopsy of the breast. Designed features of breast biopsy instrumentation are discussed. Procedures for performing breast biopsies as well as clinical indications are also explored. PMID- 7489311 TI - MR imaging of the breast after surgery for breast cancer. AB - Unenhanced scans, with any sequence, often give limited and unreliable information when trying to distinguish tumor recurrence from scarring. Dynamic scanning very soon after the intravenous injection of Gd-DTPA is the best available method of distinguishing between benign tissue and neoplastic recurrence after conservation treatment for breast cancer. 3-D T1-weighted gradient echo images appear to be the technique of choice, preferably with image subtraction. Rapid and pronounced tissue enhancement is strongly suggestive of tumor recurrence; however, scanning within 6 months of surgery and 9 months of radiation therapy may lead to false-positive results. Benign proliferative disease and recent fat necrosis may also mimic cancer on dynamic contrast enhanced MR images. The indications for MR imaging after tylectomy and radiotherapy are findings that are equivocal or suspicious of neoplastic recurrence on clinical examination, mammography, or ultrasound, often on a combination of these modalities. Typical clinical findings are a mass or induration at the site of surgery, a mass lesion with spiculated and irregular outline on mammography, or variable echogenicity on ultrasound. The development of these appearances on sequential examination is particularly suggestive of recurrence. When microcalcification is the only finding of a carcinoma on mammography after treatment for breast cancer, contrast enhancement is consistently present. PMID- 7489312 TI - MR imaging of the breast in the follow-up evaluation of conservative nonoperatively treated breast cancer. AB - Because of postradiation changes in the breast, evaluation of the efficacy of conservative local radiation treatment is difficult, and early detection of recurrent disease is often a diagnostic challenge. MR imaging of the breast aids in the follow-up of conservatively treated breast cancers. When recurrent disease is suspected, MR imaging allows for the differentiation between late postradiation changes in the breast and recurrent carcinoma. PMID- 7489313 TI - Technologic considerations for breast tumor size assessment. AB - Breast tumor staging and treatment response is primarily determined by tumor size. Presently there are four techniques to measure breast cancer size: (1) palpations, (2) mammography, (3) ultrasonography, and (4) MR imaging. This article reviews the applications and limitations of each technique and the improvements MR imaging can provide. PMID- 7489315 TI - MR imaging of breast implants. AB - MR imaging has proved to be an excellent imaging modality in locating free silicone and evaluating an implant for rupture, with a sensitivity of approximately 94% and specificity of 97%. Silicone has a unique MR resonance frequency and long T1 and T2 relaxation times, which allows several MR sequences to provide excellent diagnostic images. The most commonly used sequences include T2-weighted, STIR, and chemical shift imaging (Figs. 3, 13, and 14). The T2 weighted and STIR sequences are often used in conjunction with chemical water suppression. The most reliable findings on MR images for detection of implant rupture include identification of the collapsed implant shell (linguine sign) and free silicone within the breast parenchyma. PMID- 7489314 TI - MR mammography with pharmacokinetic mapping for monitoring of breast cancer treatment during neoadjuvant therapy. AB - The increased clinical understanding of neovascularization and its changes during therapy are the basis for therapy-response monitoring with MR mammography of breast cancer treatment. With dynamic MR imaging, objective quantification of neovascularization can be achieved using pharmacokinetic analysis of contrast media enhancement. This technique likely will be of major clinical importance for the noninvasive evaluation of neoadjuvant therapy with a favorable cost-benefit ratio. PMID- 7489316 TI - MR imaging of the breast. Imaging and tissue characterization without intravenous contrast. AB - The development of noninvasive techniques should always be an important goal of diagnostic radiology. Despite the recent excitement regarding intravenous MR imaging contrast agents (e.g., Gd-DTPA), the role of noncontrast MR imaging for the breast should not be overlooked. First, it must be kept in mind that administration of an intravenous contrast material is an invasive procedure that poses some health concern and may not be acceptable to certain patients. Second, injection of intravenous contrast media is an expensive procedure that incurs costs related to the contrast media itself as well as from the personnel required to administer the media and monitor any adverse reactions. Third, there may be specific imaging situations in which a lesion must be located prior to administration of contrast media, for example, when dynamic contrast enhanced imaging is to be performed and temporal resolution limits the volume of coverage to only a few slices. Finally, noncontrast MR imaging techniques also may be useful in their own right for obtaining information not otherwise available from a contrast-enhanced study, particularly quantitative relaxometric and spectroscopic measurements that may be useful for tissue characterization. It should also be noted that optimized noncontrast MR methods also might be used in conjunction with contrast agents to improve overall clinical utility. The heterogeneous nature of the breast and the overlap in T1 and T2 between different normal and abnormal breast tissue types frequently renders conventional noncontrast breast MR images confusing and ambiguous. Hybrid imaging approaches combining T1 weighting, T2 weighting, and fat suppression techniques as well as multivariate image analysis may be helpful for improving the sensitivity of MR imaging to breast disease; however, specificity remains a problem. Recent advances in clinical MR imaging have provided the opportunity to explore other approaches which may shed new light on the characterization of breast disease including NMRD, spin locking, magnetization transfer, and multicomponent relaxation analysis. MR imaging derived morphologic factors (i.e., lesion spiculation, parenchymal patterns) also may have a role to play in breast disease diagnosis and risk assessment. Several avenues of future development of noncontrast breast MR imaging would seem fruitful. NMRD and spin locking results suggest the potential contrast benefit of low field imaging of the breast, particularly, with the probable development of open concept low field MR imaging devices, which will provide improved access and low cost. In vivo relaxation time measurements, particularly multi-component T2 analysis and incorporation of MTC and chemical shift imaging techniques, may improve breast MR image specificity.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 7489317 TI - MR spectroscopy of the breast. AB - Breast cancer results from perturbations in normal cellular biochemistry that lead to alterations in the flux of intermediary metabolites and in the function of cell membranes. Alterations in cellular biochemistry can be measured and monitored noninvasively by MR spectroscopy, and the data so derived are complementary to MR imaging and portray interactions of biomolecules in metabolic pathways and membranes. The biochemical information obtained from MR spectra can be used to characterize and distinguish disease and nondisease states for diagnostic and therapeutic purposes. This information may be used to expand our knowledge of tumor biology and to determine the extent of disease and prognosis. MR spectroscopy, which was once the sole province of the physical chemist, is now an instrument of the physician. PMID- 7489318 TI - Breast MR imaging. An analysis of its role with respect to other imaging and interventional modalities. AB - The history of breast imaging has focused on the role of dedicated mammography, with its unique ability to provide exquisite contrast and resolution. The dual challenge of breast imaging--public health screening and diagnosis--requires unparalleled considerations of radiation dose and cost, and has prompted the development of other technologies that have failed to replace but have often defined ancillary roles to mammography. The role of breast MR imaging must be measured by these standards, and requires critical appraisal of both its unique contributions and practical limitations. PMID- 7489319 TI - Sleep, microbes and cytokines. AB - Dynamic changes in sleep in response to infectious challenge are a facet of the acute phase response. Changes in sleep induced by infection seem to be of recuperative value to the host. Furthermore, loss of sleep is associated with changes in immune function. Specific components of microbes such as muramyl peptides or endotoxin from bacteria or double-stranded RNA from virus induce sleep responses. These microbial-induced responses are mediated via enhanced cytokine and hormone production. Interleukin-1, tumor necrosis factor and interferon-alpha are somnogenic. Interleukin-1-enhanced sleep involves growth hormone-releasing hormone. Microbial-cytokine-altered sleep results from an amplification of physiological sleep mechanisms. PMID- 7489320 TI - Localization of interleukin 1 type I receptor mRNA in rat brain. AB - During states of infection and inflammation, systemic interleukin 1 (IL-1) results in clear biological effects mediated through the central nervous system, such as fever, anorexia, and activation of the hypothalamic-pituitary-adreno cortical axis. Given the multiple central effects of IL-1 in the rat, it would be expected that IL-1 receptors ought to be present in rat brain. However, no previous studies have localized IL-1 receptor or its mRNA in rat brain, possibly due to the fact that interspecies probes were used in previous studies. The recent cloning of the rat IL-1 type-I receptor (IL-1RI) has permitted us to conduct an in situ hybridization study using a species-specific, 35S-labeled anti sense riboprobe to localize IL-1 receptor mRNA in rat brain. We localized IL-1RI mRNA in the hippocampus, choroid plexus, and cerebellum. At the cellular level we found IL-1RI mRNA in low to moderate levels in hippocampal neurons and in Purkinje cells of the cerebellar cortex, and in high levels in the endothelium of postcapillary venules and in glial cells surrounding arterioles throughout the brain. This pattern of localization provides support to the concept that IL-1 crosses the blood-brain barrier. Future studies are needed to clarify how the expression of the gene encoding IL-1RI is regulated in brain tissue. PMID- 7489322 TI - Binding of anti-inflammatory alpha-melanocyte-stimulating-hormone peptides and proinflammatory cytokines to receptors on melanoma cells. AB - alpha-Melanocyte-stimulating hormone (alpha-MSH1-13), a peptide derived from proopiomelanocortin, has remarkable anti-inflammatory and antipyretic activities. This peptide and a tripeptide that forms the COOH-terminal portion of the molecule (alpha-MSH11-13; Lys Pro Val) inhibit inflammation when given centrally or peripherally. Because of the similarity in their actions, the tripeptide has been presumed to be the amino acid message sequence underlying the effects of alpha-MSH1-13. To test the possibility that the two peptides occupy the same receptors, competitive binding experiments were performed with B16 mouse melanoma cells that are known to have alpha-MSH1-13 receptors. In these experiments, alpha MSH11-13 did not inhibit binding of a radiolabelled alpha-MSH1-13 analog. This finding suggests that alpha-MSH1-13 and alpha-MSH11-13 exert their anti inflammatory/antipyretic/anticytokine effects via stimulation of separate receptors. Because alpha-MSH inhibits the effects of several cytokines including inflammation caused by interleukin (IL)-6 and IL-8, the capacity of these cytokines to compete for alpha-MSH binding sites was tested. There was no evidence that these proinflammatory cytokines bind to alpha-MSH receptors on murine melanoma cells. Although further tests with host cells involved in inflammation are required, the latter result is the first evidence that the mechanism of anticytokine action of alpha-MSH does not depend upon peptide/cytokine competition for binding sites. PMID- 7489321 TI - Inhibition of nitric oxide synthase in the hypothalamus blocks the increase in plasma prolactin induced by intraventricular injection of interleukin-1 alpha in the rat. AB - Injection of interleukin-1 (IL-1) into the third cerebral ventricle (3V) of conscious rats increases plasma prolactin (PRL) concentration. Nitric oxide (NO) is involved in control of corticotropin-releasing factor (CRF) and luteinizing hormone-releasing hormone (LHRH) release. Consequently, we evaluated its role in the PRL-releasing action of IL-1. In the present experiment, NG-mono-methyl-L arginine (NMMA) (1 mg in 5 microliters of 0.9% NaCl (saline)], an inhibitor of NO synthase, or 5 microliter of saline was microinjected into the 3V of conscious, castrate male rats and blood samples were removed from jugular catheters just prior to and at 10-min intervals after injection. A second injection of NMMA or saline was given 60 min after the first. Five minutes after the injection of NMMA or saline, IL-1 alpha (0.6 pmol in 2 microliter saline), or an equal volume of saline, was injected into the 3V. Plasma PRL concentrations were increased within 10-20 min after injection of IL-1 alpha and a second pulse of PRL usually occurred at 60-70 min following its injection. The maximal increase in plasma PRL from the initial value in the animals injected with IL-1 alpha was highly significant, whereas there was no significant increase in the animals injected with NMMA plus IL-1 alpha or in the animals injected with saline or NMMA. The area under the plasma PRL curve was significantly elevated in the animals injected with IL-1 alpha above that of rats injected with NMMA plus IL-1 alpha during the first hour after injection.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7489324 TI - Corticotropin-releasing factor microinfused into the locus coeruleus produces electrocortical desynchronization and immunosuppression. AB - In freely moving rats with cannulae chronically implanted into the locus coeruleus (LC), the effects of corticotropin-releasing factor (CRF) on electrocortical (ECoG) spectrum power activity and on immune mechanisms (splenocyte mitotic response to concanavalin A, Con A, and lipopolysaccharide, LPS, natural killer cell, NK, activity) were assessed. CRF (100-300 ng) microinfused into the LC produced marked ECoG desynchronization characterized by a significant decrease in total voltage power as well as in power of frequency bands of 0.25-3 and 3-6 Hz. These effects lasted 30-60 min according to the dose. A prior administration of alpha-helical CRF(9-41) (200 ng into the LC 15 min before) prevented ECoG desynchronization induced by CRF (100 ng). In addition, CRF (100 ng) given into the LC produced a significant decrease 45 min later in the splenocyte proliferative response to Con A and LPS and a significant fall of NK activity. These effects were prevented by prior microinfusion into the same site of alpha-helical CRF (200 ng). In conclusion, the present experiments show that CRF given into the LC produces an intense state of ECoG desynchronization accompanied by marked immunodepression and suggest that LC is an important site in the brain through which CRF exerts its immunosuppressive activity. PMID- 7489325 TI - A reappraisal of the role of the various opioid receptor subtypes in cell mediated immunity. AB - Opioid peptides have been shown by several studies to modulate various parameters of the immune response, but scant experimental findings exist on the role played by specific opioid receptor subtypes in the control of immune mechanisms. This study focuses on the in vitro influences of [Trp4,Asn7]dermorphin, a mu-selective agonist, [D-Ala2]deltorphin I, a delta-selective agonist and U50,488, a kappa selective agonist, on the proliferative response of splenocytes to concanavalin A (Con A). [Trp4,Asn7]dermorphin at low concentrations (10(-11P) and 10(-12) M) enhanced the proliferative response to Con A, whereas higher concentrations (10( 6) to 10(-7) M) inhibited it. Both effects were antagonized by naloxone. [D Ala2]deltorphin I at very low concentrations (10(-12) to 10(-13) M) also produced a significant increase in the proliferative response of splenocytes to Con A. This effect was significantly antagonized by natrindole, a specific delta receptor antagonist. Finally U50,488 at concentrations ranging from 10(-8) to 10( 9) M inhibited the proliferative response to Con A. The effects of U50,488 were mediated by the stimulation of the kappa-opioid receptors, since a preincubation of splenocytes with the selective antagonist norbinaltorphimine significantly reduced or abolished the U50,488-induced suppression of the mitotic response. In conclusion, our results clearly indicate that the different opioid receptor subtypes play a different role in the control of immune mechanisms and suggest that immunoenhancing effects of opioid peptides are very likely due to the stimulation of mu- and delta-receptors, whereas the immunosuppressive effects are mediated through the stimulation of kappa-opioid receptors. PMID- 7489323 TI - Rapid increase in plasma IL-6 after hemorrhage, and posthemorrhage reduction of the IL-6 response to LPS, in conscious rats: interrelation with plasma corticosterone levels. AB - To study whether hemorrhage stimulates interleukin-6 (IL-6) production in conscious rats, 30% of the total blood was withdrawn over 3 min through an indwelling venous catheter and the shedblood was reinfused 1 h later. Plasma adrenocorticotropic hormone (ACTH), corticosterone and IL-6 concentration rapidly increased. Plasma ACTH levels peaked at 10 min and corticosterone and IL-6 peaked at 60 min; all started to decrease after reinfusion. In adrenalectomized (ADX) rats with or without a corticosterone pellet implant, there was an inverse relationship between IL-6 and corticosterone concentrations, greatest in ADX rats and lowest in ADX rats in which plasma corticosterone was elevated by crushing the implanted pellet. However, the ADX rats in which plasma corticosterone was maintained at normal or slightly elevated levels showed greater IL-6 responses to hemorrhage and elevated basal plasma IL-6 levels compared to sham-operated control rats. Twenty-four hours after hemorrhage/reinfusion, ACTH, corticosterone, and IL-6 responses to i.v. injection of lipopolysaccharide (LPS) were all reduced compared to the non-hemorrhaged animals, indicating that hemorrhage impaired general host defense. Although very high plasma corticosterone concentrations markedly suppressed the IL-6 response to LPS, in ADX rats in which plasma corticosterone was maintained at slightly higher levels than normal, the reduced IL-6 response to LPS in the posthemorrhage period was not reversed, but enhanced. Thus corticosterone has biphasic effects on the IL-6 response to hemorrhage and the response to LPS during the posthemorrhage period, which has important clinical implications with regard to the optimal dose of glucocorticoid for maintaining the host defense response.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7489326 TI - The neuropeptide alpha-melanocyte-stimulating hormone: a key component of neuroimmunomodulation. AB - Recent research indicates that the proopiomelanocortin derivative alpha melanocyte stimulating hormone (alpha-MSH) is a significant modulator of host reactions including fever and inflammation. Although the precise mechanism of action is still unknown, cytokine antagonism is believed to be responsible for at least a part of its anti-inflammatory/antipyretic influence: alpha-MSH antagonizes pyrogenic and proinflammatory effects of cytokines such as interleukin-1 (IL-1), interleukin-6 (IL-6), tumor necrosis factor (TNF), and interferon gamma (IFN gamma). Although it is clear that the peptide can act within the brain to inhibit fever and peripheral inflammation, an anti inflammatory effect on a peripheral target was evidenced in animals with transection of the spinal cord. Recent data show that alpha-MSH is significant also in human disorders such as AIDS, rheumatoid arthritis, and myocardial infarction. This molecule is believed to be a key factor in neuroimmunomodulation and it may be useful as a therapeutic agent in control of inflammatory reactions. PMID- 7489327 TI - Hypothalamic-pituitary-thyroid axis and the immune system. AB - The paper reviews data on bidirectional circuits between the hypothalamic pituitary-thyroid (HPT) axis and the immune system. The effects of thyroliberin (TRH), thyrotropin (TSH) and of thyroid hormones (thyroxine and triiodothyronine) on the immune system, as well as the effects of mono- and lymphokines on the HPT axis are discussed. PMID- 7489329 TI - Stimulation of transcriptional regulatory activity by substance P. AB - The neuropeptide substance P (SP) is one of the principal mediators of neurogenic inflammation as well as a neurotransmitter in nociceptive affect neurons. The mechanisms by which binding of SP to its receptor stimulates diverse downstream biologic effects remain unknown. In order to elucidate this process we have established stably transfected cell lines expressing functional rat SP receptors (KNRK-SPR). When stimulated by SP, KNRK-SPR cells respond by simultaneously mobilizing intracellular Ca2+ and increasing cAMP levels. To determine if SP stimulation activates downstream transcriptional regulatory factors, we transfected KNRK-SPR cells with plasmids containing the activator protein 1 (AP 1) and cAMP-responsive (CRE) enhancer elements coupled to the chloramphenicol acetyltransferase (CAT) reporter gene. Stimulation with SP 1-1,000 nM caused a 1.5- to 2-fold increase in CAT activity in both AP-1-CAT- and CRE-CAT-transfected KNRK-SPR cells. Northern and Western blot analyses demonstrate that the mechanism by which SP stimulates AP-1 enhancer activity involves increases in both c-jun mRNA and protein. Moreover, gel retardation assays with oligomers containing the AP-1 and CRE binding sites showed that SP induces specific retardation bands consistent with increases in AP-1 and CRE complexes. These experiments suggest that SP-mediated stimulation of cells involves the participation of two signaling pathways resulting in several transcriptional regulatory mechanisms being activated. PMID- 7489328 TI - Alpha-melanocyte-stimulating hormone inhibits corticotropin-releasing factor release by blocking protein kinase C. AB - Cytokine-induced release of corticotropin-releasing factor (CRF) from hypothalamic explants in vitro can be inhibited by femtomolar concentrations of alpha-melanocyte-stimulating hormone (alpha-MSH). Because the mechanism of the anticytokine action of alpha-MSH remains unknown, we examined if the peptide inhibits CRF release by interference with various steps in the activation of CRF release. Previous studies have shown that CRF release is induced by activation of phospholipase A2 (PLA2). Therefore, we examined the effect of alpha-MSH on the action of melittin (MEL), a PLA2 activator. After 60 min preincubation in Krebs Ringer bicarbonate buffer, medial basal hypothalami were incubated for 30 min with Krebs-Ringer bicarbonate buffer or MEL with or without alpha-MSH (10(-11) to 10(-16) M). CRF release into the incubation medium was measured by RIA. As reported previously none of the alpha-MSH concentrations used changed basal CRF release nor did any concentration of alpha-MSH significantly alter CRF release induced by MEL (10 micrograms/ml). Thus, alpha-MSH alters cytokine-induced CRF release at a step unrelated to the activation of PLA2. Because activation of PLA2 requires an increase in intracellular calcium ion (Ca2+) concentrations, we evaluated the effect of alpha-MSH on the release of CRF induced by a high concentration of potassium (56 mM). This concentration of potassium induced a 3.5 fold increase in CRF release that was not affected by alpha-MSH. Protein kinase C (PKC) stimulates CRF release. Consequently, we examined the effect of alpha-MSH on CRF release induced by phorbol myristate acetate (PMA), which in the presence of Ca2+ stimulates PKC.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7489330 TI - Brain regulatory system for the immune response: immunopharmacology and morphology. AB - On the basis of our previous investigations concerning the influence of some central nervous system structures on the immune reaction, we have shown the existence of a brain regulatory system for immune response (BRSIR). We have investigated this BRSIR using small electrolytic lesions placed in different brain areas of male rats, followed by two methods to evaluate the effect of the lesions on the immune response, the method of delayed skin hypersensitivity and the method of utilization of 3H-thymidine for synthesis of DNA after injection of muramyl dipeptide, a very potent immunostimulating compound. The experiments were performed on rats of the Wistar strain. Lesions were placed from the spinal cord through the brain stem to the cerebral cortex. The results suggest that the following structures constitute parts of the BRSIR: medial frontal cortex (areas Cg 1-3), subnucleus basomedialis and centralis of the amygdala, subnucleus medialis and dorsolateralis of the nucleus parabrachialis, lateral reticular formation (nucleus parvocellularis--mainly areas corresponding with aminergic groups A1-7), part of the raphe reticular formation (nucleus raphealis dorsalis and nucleus linearis--mainly areas corresponding with serotoninergic groups B6 8), and the spinal cord. PMID- 7489331 TI - Evidence for an involvement of dopamine D1 receptors in the limbic system in the control of immune mechanisms. AB - Indirect evidence suggests that dopamine within the brain may participate in the regulation of immune responses both in humans and in rodents. The aim of the present study was to investigate the possible modulatory role played by specific dopamine D1 receptor subtypes within discrete sites of the brain dopaminergic pathways. Mitogen responsiveness of splenocytes and natural killer (NK) cell activity were measured in rats following microinfusion of SKF 38393, a specific dopamine D1 receptor agonist, into the ventral tegmental area, the amygdala, the nucleus accumbens and the CA1 area of the hippocampus. We report here that microinfusion of SKF 38393 (100 nmol) within the central amygdala increased the proliferative response of splenocytes to concanavalin A (ConA) while it did not modify the proliferative response of splenocytes to lipopolysaccharide (LPS) or NK cell activity. On the contrary, microinfusion of SKF 38393 into the nucleus accumbens decreased the proliferative response of splenocytes to ConA and LPS, while NK cell activity remained unchanged. Similarly, microinfusion of SKF 38393 into the CA1 area of the hippocampus decreased the proliferative response of splenocytes to LPS, but not to ConA and did not affect NK cell activity. Finally, microinfusion of SKF 38393 into the ventral tegmental area did not significantly modify the proliferative response of splenocytes to either ConA or LPS and did not affect NK cell activity. All immunological changes evoked through the different areas of the brain following microinfusion of SKF 38393 were prevented by systemic administration of SCH 23390, a specific D1 receptor antagonist. PMID- 7489332 TI - Localization of stem cell factor mRNA in adult rat hippocampus. AB - Stem cell factor (SCF) promotes the growth of multilineage hematopoietic cells. SCF is a product of the steel (Sl) locus of the mouse, and it is a ligand for the c-kit proto-oncogene receptor. Previous studies have investigated the distribution of SCF mRNA in developing and adult tissues of the rat, including the brain. However, there have been conflicting reports on the distribution of SCF mRNA in adult rat brain. Specially noteworthy was one report of the absence of SCF mRNA in adult hippocampus, while another group reported the presence of that mRNA in the dentate gyrus of the hippocampus. We conducted this study to determine the precise localization of SCF mRNA in adult brain, and were especially interested in determining whether that mRNA is localized in adult hippocampus. We used in situ hybridization histochemistry to demonstrate that the gene encoding SCF is actively expressed in neuron-like cells in various regions of adult rat brain. Our data show that SCF mRNA is present in neuron-like cells in the thalamus, cerebral cortex, cerebellum, and hippocampus, particularly in the dentate gyrus, but also in CA1, CA2, and CA3. We did not localize SCF mRNA in glia-like cells. Dyskeratosis congenita is a severe human disorder, associated with dyskeratosis, anemia, and mental retardation. It has been postulated that dyskeratosis congenita is due to a deficiency in SCF function. It is unknown why patients with dyskeratosis congenita suffer from mental retardation.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7489333 TI - Alpha-melanocyte-stimulating hormone suppresses antigen-stimulated T cell production of gamma-interferon. AB - The neuropeptide alpha-melanocyte-stimulating hormone (alpha-MSH) is known to suppress cytokine-mediated inflammation. In addition, we previously found that alpha-MSH suppressed the production of the proinflammatory cytokine interferon (IFN)-gamma by antigen-stimulated primed lymph node T cells. This immunosuppressive activity of alpha-MSH on lymph node T cell cultures is similar to that of interleukin (IL)-4. To further examine the potential 'IL-4 like' activities of alpha-MSH, antigen-stimulated lymph node T cell cultures were treated with alpha-MSH in the presence of neutralizing anti-IL-4 antibodies. The enhanced production of IFN-gamma caused by the presence of anti-IL-4 alone in the T cell cultures was squelched by alpha-MSH. This demonstrated that in these cultures, alpha-MSH regulation of IFN-gamma production operates in a fashion similar to that of endogenous IL-4. Addition of exogenous IL-4 to antigen stimulated lymph node T cell cultures did not intensify alpha-MSH down-regulation of IFN-gamma production, and the addition of alpha-MSH to IL-4-treated cultures did not further depress IFN-gamma production. These and the previous results suggest that the mechanism of alpha-MSH suppression of IFN-gamma production in the antigen-stimulated T cell cultures is similar to, but independent of, IL-4. When antigen-presenting cells (APCs) were the only cells in the antigen stimulated T cell cultures treated with alpha-MSH, there was a significant reduction (60-70%) of APC elicitation of IFN-gamma production by untreated primed T cells.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7489334 TI - Role of nitric oxide in control of growth hormone release in the rat. AB - Previous experiments in this and other laboratories have revealed that nitric oxids (NO) plays a role in controlling the release of corticotropin-releasing hormone (CRH) and luteinizing-hormone-releasing hormone (LHRH). Therefore, we have investigated its role in control of growth hormone (GH) release in conscious rats by microinjecting NG-monomethyl-L-arginine (NMMA), an inhibitor of NO synthase (NOS), into the third ventricle (3V) of conscious, freely moving castrate male rats. An initial blood sample (0.3 ml) was drawn from an indwelling intra-atrial catheter just prior to injection of NMMA [1 mg in 5 microliters of 0.9% NaCl (saline)] into the 3V. To maintain the inhibitory action on NOS, a second injection of NMMA was administered into the 3V 60 min after the first. Additional blood samples (0.3 ml) were removed at 10 min intervals for 120 min. Other animals received injections of the diluent at the same times and volumes as NMMA. Interleukin (IL)-1 alpha (0.06 pmol in 2 microliters saline) was injected into the 3V immediately after the first injection of NMMA, whereas other animals received the NMMA diluent followed by IL-1 alpha. The effects of IL-1 alpha were almost identical to those of NMMA in that there was a dramatic lowering of plasma GH achieved primarily by a reduction in height of the GH pulses without a significant reduction in their number.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7489335 TI - The role of the growth and lactogenic hormone family in immune function. AB - The influence of various members of the growth and lactogenic hormone family on the immune system is reviewed. A general hypothesis is proposed for growth control in higher animals. It is suggested that immune reactions, which are based on lymphocyte proliferation, obey the general rules of growth control in vertebrate animals. Growth and lactogenic hormones (GLH) are required for the development and function of the immune system and are suggested to deliver the first signal that prepares the cell for proliferation, differentiation and function. This signal has already been designated by other as the competence signal which initiates the cell cycle. Second signals are delivered through the antigen receptor, and/or by some other cell surface receptors (adhesion molecules) and always involve cell-to-cell ('bridging') and/or cell-to-matrix interaction. This category of signals is designated as stromal or adherence signals. The lymphocyte adhesion molecules that mediate second signals have evolved form organ- and tissue-specific recognition/regulatory molecules. The antigen receptors have been perfected during evolution from self recognition to specific-antigen recognition. Apart from this exquisitely specific mechanism of immune recognition, there is evidence for other less specific means of recognition by adherence molecules that mediate the activation of the immune system during nonspecific injury and also play a role in the elimination of degenerated and neoplastic cells. Signals delivered through adhesion molecules have the power to commit the cell to a given activity which is executed by the delivery of third signals in the form of soluble cytokines, usually, but not always, by the same cell delivering the second signal(s). The combination of these three groups of signals will ultimately determine whether or not the cell will proliferate, differentiate, maintain function or, perhaps, be committed to apoptosis. Therefore, GLH maintain immunocompetence which enables the immune system to respond to specific antigenic and tissue-derived stimuli in a self regulated fashion. The adrenocorticotropic hormone-adrenal axis antagonizes the immunostimulatory effect of GLH. This basic pattern of lymphocyte regulation is influenced further by additional hormones, neurotransmitters and neuropeptides, mostly by the modulation of signal delivery. The constant interaction of neuroendocrine and internal immunoregulatory mechanisms assures the fine tuning of the immune system, so that it is able to function in homeostasis and harmony with the organism. PMID- 7489336 TI - Neuronal factors modulating immunity. AB - In this minireview we will discuss some evidence suggesting that the immune response is under neuronal regulation. In particular, we will concentrate on the effects that various neuropeptides have on immunity both in vitro and in vivo. Of these, vasoactive intestinal peptide, substance P, somatostatin, and calcitonin gene related peptide will be discussed in detail. In addition, the effects of nerve growth factor on the immune system will be presented. Finally, a possible role for these neuropeptides in various diseases and its clinical relevance will be suggested. PMID- 7489337 TI - The estrous cycle and pituitary-ovarian function in Lewis and Fischer rats. AB - We have characterized the estrous cycle by obtaining vaginal smears, and quantitating estradiol (E), progesterone (P), luteinizing hormone (LH), and follicle-stimulating hormone (FSH) plasma levels at different phases of the estrous cycle in Lewis (LEW/N) and Fischer (F344/N) rats. Comparison of the duration of the component phases of estrous showed LEW/N metestrous to be significantly longer than in F344/N rats while diestrous and estrous were significantly shorter; proestrous was identical. E levels in LEW/N rats were significantly greater than in F344/N rats only in the estrous phase of the cycle. P levels were significantly greater in LEW/N rats in all phases. LH and FSH levels in the two strains did not differ. Elevated E and P levels would be expected to be associated with increased corticosterone through inhibition of the glucocorticoid negative-feedback pathway. The data reported suggest that other modulating factors in corticotropin-releasing-factor synthesis/release could be overriding both the E and P effects upon hypothalamic-pituitary-adrenal axis responsiveness. PMID- 7489339 TI - Pulsatility of 24-hour concentrations of circulating interleukin-1-alpha in healthy women: analysis of integrated basal levels, discrete pulse properties, and correlation with simultaneous interleukin-2 concentrations. AB - Even though it is widely known that interleukin (IL)-1 alpha acts at the local level, it is still uncertain whether IL-1 alpha is secreted into the circulation and acts at distant sites. We have tried to elucidate this by measuring 24-hour levels of total IL-1 alpha in six healthy female volunteers. Subjects had detectable and pulsatile levels of IL-1 alpha throughout the 24-hour period. The integrated 24-hour IL-1 alpha concentration was 2,367 +/- 753 min x micrograms/l (mean +/- SD), and the integrated pulsatile IL-1 alpha concentration was 553 +/- 260 (25 +/- 10% ot total integrated IL-1 alpha). The mean IL-1 alpha concentration was 1.63 +/- 0.53 micrograms/l, mean pulse frequency/24 h was 12.8 +/- 0.8, mean pulse height was 2.31 +/- 0.52 micrograms/l; mean pulse width was 80.4 +/- 2.3 min, and mean interpulse interval was 105.3 +/- 2.8 min. Total IL-1 alpha levels significantly correlated with those previously reported for IL-2 in the same samples, and IL-1 alpha pulse parameters which are concentration independent were significantly similar to those of IL-2. Furthermore, cross correlation analysis indicated that in 83% of our subjects (5/6) there was synchronicity of IL-1 alpha and IL-2 levels. IL-1 alpha pulse parameters were in the range reported for hormones which have well-characterized pulsatility, such as growth hormone, luteinizing hormone, follicle-stimulating hormone, cortisol, beta-endorphin, and progesterone. Based on these data we speculate that a pulsatile cytokine cascade may exist in the systemic circulation.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7489338 TI - kappa-Opioid receptor functions: possible relevance to experimental allergic encephalomyelitis. AB - The present study deals with the influence of preferential kappa-opioid agonist MR 2034 on experimental allergic encephalomyelitis (EAE). For this purpose, 9 week-old male Dark August rats were treated intraperitoneally with 0.2 mg/kg of MR 2034 as follows: (a) from the day of EAE induction until sacrifice; (b) from the day of EAE induction until the appearance of neurological signs, and (c) from the appearance of neurological signs until sacrifice. Repeated injections of MR 2034 given during the whole period of observation produced the most pronounced suppression of EAE clinical signs, histological lesions in the brain and spinal cord, and anti-myelin basic protein antibody production. These results suggest that kappa-opioid receptors may be involved in the development of EAE. PMID- 7489340 TI - An interleukin-1-alpha-like neuronal system in the preoptic-hypothalamic region and its induction by bacterial lipopolysaccharide in concentrations which alter pituitary hormone release. AB - We studied the effect of intravenous injection of lipopolysaccharide (LPS) (30 250 micrograms) on the release of several anterior pituitary hormones as indicated by changes in their concentrations in plasma. Within 30 min after intravenous injection of LPS there was a dose-related stimulation of ACTH release; prolactin (PRL) release was induced only by the highest LPS dose injected (250 micrograms). Even the lowest dose of LPS (30 micrograms) decreased plasma growth hormone (GH) by 60 min. Higher doses lowered plasma GH by 30 min, but thyroid-stimulating hormone release was only significantly inhibited by the highest dose of LPS. The action of LPS seems to be primarily exerted on the central nervous system, since incubation of hemipituitaries with LPS for 3 h in doses ranging from 0.001 to 10 micrograms/ml had no effect on ACTH release. LPS is thought to induce its effects on hormones either by release of cytokines from immune cells which subsequently induce the hormonal changes or possibly by direct action within the hypothalamus. In this report we demonstrate the immunocytochemical localization of a population of interleukin-1 alpha (IL-1 alpha)-like cells in a region extending from the basal forebrain at the level of the diagonal band of Broca, caudally and dorsally to the dorsolateral preoptic region and the hypothalamus at the level of the paraventricular nucleus. Further caudally, IL-1 alpha-like immunoreactive cells were located in the midportion of the amygdala. Two hours after injection of the 125-micrograms dose of LPS, the number of these immunoreactive cells was dramatically increased.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7489341 TI - Muscarinic cholinergic receptors of rat lymphocytes: effect of antigen stimulation and local brain lesion. AB - Expression of muscarinic cholinergic receptors (m-AchR) on thymocytes and lymphocytes supports a hypothesis of direct interaction between autonomic innervation and the immune system. Using a muscarinic antagonist, [3H]quinuclidinyl benzilate ([3H]QNB), we revealed different characteristics of m AchR expression in lymphocytes isolated from the spleen, peripheral blood or thymus of Wistar rats. To further explore the mechanisms controlling m-AchR expression in lymphocytes we have addressed two questions: (i) whether local brain lesions will affect [3H]QNB binding activity of lymphocytes and (ii) whether different antigens will alter m-AchR expression in lymphocytes. We report here that electrolytic lesions of the area hypothalamica anterior resulted in a significant increase in [3H]QNB-specific binding in thymocytes 7 days after neurosurgery. In contrast, local lesions of other hypothalamic structures (area preoptica medialis, area hypothalamica posterior) or sensomotor cortex did not affect m-AchR expression in thymocytes. No apparent changes of [3H]QNB-binding activity were found in lymphocytes isolated from rat spleen or peripheral blood after operation. Both T-cell-dependent (sheep red blood cells) and T-cell independent (Vi antigen of Salmonella typhi) antigens induced a significant increase in [3H]QNB-specific binding in spleen lymphocytes while [3H]QNB-binding activity of peripheral blood lymphocytes did not change. A different pattern of lymphocyte m-AchR expression found in various lymphoid tissues after immunization or brain lesions suggests a local involvement of cholinergic mechanisms in neuroimmune interaction. PMID- 7489342 TI - The structure of hepatitis B envelope and molecular variants of hepatitis B virus. AB - Accumulated evidence in recent years has shown that the variation of hepatitis B virus (HBV) genomes may have profound implications for our understanding of hepatitis B pathogenesis and prevention. Attention has focused on areas of the outer envelope coded by the S gene which are involved in the induction of a protective neutralising antibody response, and mutations which directly affect the production of C gene products, one of which is considered as a target for immune T cells involved in virus clearance. This review highlights recent experimental data which emphasizes the role of such mutations in the establishment and maintenance of chronic HBV infections and focuses attention on the significance of HBV variants with respect to the expanding use of HBV vaccines for mass immunization. PMID- 7489343 TI - The fluctuations of hepatitis C virus RNA and IgM anti-HCV (core) serum levels correlate with those of alanine aminotransferases during the hepatitis relapses of patients treated with interferon. AB - Variations in the serum levels of hepatitis C virus (HCV) RNA. IgM antibody against the HCV 'core' structural protein (c22) and alanine amino-transferase (ALT) were measured in 23 patients with chronic hepatitis C who underwent therapy with interferon-alpha 2a (IFN alpha 2a). Low pretreatment levels of viraemia and undetectable IgM anti-core were significantly associated with a long-term response to treatment. In patients with hepatitis relapses after the end of treatment, HCV RNA levels increased before or at the same time as ALT in 29 out of 34 cases (85%). ALT flares occurred before or simultaneously with IgM anti core elevations in 18 out of 20 cases (90%). Therefore, post-treatment hepatitis C exacerbations show the same sequence of events seen as in hepatitis B exacerbations (increases of viraemia followed by those of ALT and IgM anti 'core'). These findings underscore the diagnostic and prognostic usefulness of monitoring anti-HCV-positive patients with quantitative assays for HCV markers. PMID- 7489344 TI - Absence of anti-LKM-1 antibody in hepatitis C viral infection in the United States of America. AB - Several studies from Europe have observed a relationship between hepatitis C virus infection and anti-liver/kidney microsome-1 (anti-LKM-1) positive chronic hepatitis. It has been suggested that hepatitis C may induce an autoimmune phenomenon that leads to the development of a specific type (type II anti-LKM-1 positive) autoimmune chronic hepatitis. We evaluated 204 sera from patients with well-documented hepatitis C infection from two centres in the United States of America and compared them with sera from 428 French patients from three centres. We evaluated the serological prevalence of anti-smooth muscle antibodies, anti nuclear antibodies, anti-liver cytosol antibodies, and anti-mitochondrial antibodies subtype anti-M2 in patients with chronic hepatitis C. The two groups were matched in their ages, gender, mode of transmission of hepatitis C infection and severity of liver disease. Anti-LKM-1 was not observed in the patients from the USA at a time when it was noted in 3.7% of French patients. There were no differences, however, in the expression of other auto-antibodies, which were often in low titres. Absence of anti-LKM-1 in USA sera in comparison with French sera suggests that there may be differences in induction of anti-LKM-1 related to environmental and/or host genetic factors, and/or genomic variation in the hepatitis C virus. PMID- 7489345 TI - Comparative study of two high doses of lymphoblastoid interferon in the treatment of chronic hepatitis C: influence on the levels of ALT, viraemia and histologic activity. AB - Ninety consecutive patients with chronic hepatitis C were included in a randomized, uncontrolled trial to compare the efficacy of two treatment regimens, 10 MU (group A) vs 5 MU (group B), of lymphoblastoid interferon, in a step-down schedule for 24 weeks. All of the patients had antibodies against the hepatitis C virus, and all but one were HCV RNA positive in serum. The origin of the infection was attributed to blood transfusion in 30 patients and classified as sporadic in 60 patients. During treatment reduction in the ALT levels as well as the elimination of viraemia was observed in both treated groups, although these changes did not correlate significantly with the interferon dose. Nine months after the end of therapy, a sustained response was achieved in 13.6% (12/88) of the patients. Relapse in group B (87.5%) was significantly higher than in group A (59.1%). The percentage of cases which remained with undetectable HCV RNA was significantly higher for the sustained responders (66.7%) than for the non responders (11.8%) and relapser patients (2.4%). Repeated liver biopsies showed an overall significant reduction of all the subindices of histological activity from patients with sustained response, except for fibrosis. In short: the 10 MU dosing regimen of lymphoblastoid interferon was as efficient as the 5 MU dose as it brought about a similar improvement in ALT levels, histological activity and elimination of viraemia, albeit 10 MU proved significantly more effective in the prevention of a relapse among the responders after 24 weeks therapy. PMID- 7489346 TI - Contribution of HEV and HCV in causing fulminant non-A, non-B hepatitis in western India. AB - During 1990, 38 patients with fulminant non-A, non-B hepatitis (NANB) died in Government Medical College Hospital, Aurangabad. Serum samples from these patients were tested for antibodies to hepatitis C virus (anti-HCV) and IgM antibodies to hepatitis E virus (IgM-anti-HEV). All samples were also subjected to polymerase chain reaction (PCR) for the detection of HBV DNA, HCV RNA and HEV RNA. None of the patients had circulating anti-HCV antibodies; three had HCV RNA. Based on anti-HEV-IgM positivity 14 patients (37%) could be diagnosed as suffering from hepatitis E. None was positive for HEV RNA. In the absence of serological markers, HBV DNA was present in three cases. None of the HBV DNA positive patients had anti-delta antibodies. Dual infections (HBV with HEV, and HBV with HCV) were seen in two cases. The aetiology of half of the NANB cases could not be assigned to the known hepatitis viruses using current techniques. PMID- 7489348 TI - Hepatitis C virus genotypes in Singapore and Indonesia. AB - 5' untranslated and partial core (C) region sequence of hepatitis C virus (HCV) in 21 Singaporean and 15 Indonesian isolates were amplified by reverse transcription polymerase chain reaction and sequenced with the use of conserved primer sequences deduced from HCV genomes identified in other geographical regions. The HCV genotypes are predominantly that of Simmonds type 1 and less of type 2 and 3 with the latter genotype currently not detected in Indonesia. The 5' untranslated sequences are related to HCV-1. DK-7 (Denmark), US-11 (United States of America), HCV-J4, SA-10 (South Africa), T-3 (Taiwan), HCV-J6, HCV-J8, Eb-1 and Eb-8. When compared with the prototype HCV-1, insertions are found within the 5' untranslated region of Singaporean isolates and not in the Indonesians. There are Singaporean and Indonesian isolates that have sequences within the 5' untranslated region that differ slightly from each other. Microheterogeneity is observed in the core region of two Singaporeans and one Indonesian isolate. Finally, not all HCV isolates can be amplified with the conserved core sequence primers when compared with the ease with which these isolates can be amplified with 5' untranslated region conserved primers. PMID- 7489347 TI - Prevalence of hepatitis C antibody in patients with chronic liver disease and hepatocellular carcinoma in Korea. AB - To investigate the contribution of hepatitis C virus (HCV) to chronic liver disease and hepatocellular carcinoma (HCC) in Korea, antibodies to HCV (anti-HCV) were tested by enzyme immunoassay in 1759 patients with chronic liver disease and HCC, and in 808 healthy adults. The prevalence of anti-HCV was 1.6% in 808 controls. Anti-HCV was present in 32 (7.7%) of 418 hepatitis B surface antigen (HBsAg)-positive and 128 (53.1%) of 241 HBsAg-negative patients with chronic hepatitis, 16 (6.0%) of 265 HBsAg-positive and 90 (30.5%) of 295 HBsAg-negative patients with liver cirrhosis, and 16 (4.8%) of 330 HBsAg-positive and 61 (29.0%) of 210 HBsAg-negative patients with HCC. Antibodies to hepatitis B core antigen (anti-HBc) were present in 80-88% of patients who were seropositive for anti-HCV and seronegative for HBsAg. Among the sera from 114 patients with HBsAg-negative and anti-HCV-positive chronic liver diseases, HBV DNA and HCV RNA were detected by polymerase chain reaction (PCR) in 54 (47.4%) and 61 (53.3%), respectively. Both HBV DNA and HCV RNA were detected in 4 (4.4%) samples. The mean age of the patients with both HBsAg and anti-HCV was not different from that of patients who were seropositive for HBsAg alone. These findings indicate that current and/or past HBV infection is still the main cause of chronic liver disease in Korea.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7489350 TI - Detection of anti-hepatitis C virus using chemiluminescence. AB - To determine if chemiluminescence can be used in an enzyme immunoassay (EIA) format to test for hepatitis C virus (HCV) antibody and to compare sensitivity and specificity of chemiluminescence to a licensed anti-HCV EIA method, random volunteer donor samples were evaluated. One thousand and seventy-four random volunteer donor samples were collected during a 2-week period. Two aliquots of each sample were tested for anti-HCV. One aliquot was tested using a licensed anti-HCV 2.0 EIA test. The second aliquot was tested using the research anti-HCV chemiluminescence assay. Confirmatory testing was done using Recombinant Immuno Blot Assay (RIBA) 2.0 HCV and HCV RNA. Of the 1074 samples, eight were found to be reactive for anti-HCV. Seven were positive by the anti-HCV 2.0 EIA and the chemiluminescence system. The eighth sample was chemiluminescence reactive, but EIA negative; when tested by an unlicensed EIA 3.0 HCV test, RIBA 3.0 HCV test and an HCV-RNA assay, the sample was still negative. The same sample was also tested by RIBA 2.0 HCV and showed a reactive band to SOD. The chemiluminescence assay has a 100% sensitivity and 99.9% specificity compared to EIA and can be used as an alternative to EIA for detecting antibodies to HCV. PMID- 7489349 TI - Plasma glutathione concentration in patients with chronic hepatitis C virus infection. AB - It has recently been proposed that a depletion of glutathione (GSH) may be a contributing factor to viral persistence and resistance to interferon-alpha (IFN alpha) therapy in chronic hepatitis C virus (HC) infection. The aim of this study was: (1) to compare plasma GSH levels in patients with chronic HCV infection and normal healthy controls; and (2) to correlate GSH levels with liver histology and serum HCV RNA levels. Twenty-four patients with compensated chronic hepatitis C and 27 healthy subjects were studied. Serum and heparinized plasma were prospectively prepared and frozen within 1 h of collection. Plasma glutathione and glutathione peroxidase (GP) levels were measured spectrophotometrically. The serum HCV RNA level was quantitated by the branched chain DNA signal amplification assay. Plasma GSH levels were not decreased in patients with chronic HCV infection but were actually greater than in controls (control 1.27 +/ 0.12 micrograms ml-1, HCV 1.62 +/- 0.11 micrograms ml-1, P < 0.05). There was also no difference in plasma GP activity between these two groups (control 0.233 +/- 0.007 U ml-1, HCV 0.230 +/- 0.007 U ml-1). Among the patients with chronic HCV infection, there was no correlation between either plasma GSH or GP levels and the serum alanine aminotransferase (ALT) or aspartate aminotransferase (AST), serum HCV RNA level, or liver histology. This study demonstrates that chronic HCV infection does not decrease the plasma GSH and GP levels. PMID- 7489352 TI - The dark side of glucose. PMID- 7489351 TI - Targeting TGF-beta for treatment of disease. PMID- 7489353 TI - Constraining the cell cycle: regulating cell division and differentiation by gene therapy. PMID- 7489354 TI - Food for thought. PMID- 7489355 TI - The effect of increased salt intake on blood pressure of chimpanzees. AB - A colony of 26 chimpanzees given a fruit and vegetable diet of very low Na and high K intake were maintained in long-standing, socially stable small groups for three years. Half of them had salt added progressively to their diet during 20 months. This addition of salt within the human dietetic range caused a highly significant rise in systolic, mean and diastolic blood pressure. The change reversed completely by six months after cessation of salt. The effect of salt differed between chimpanzees, some having a large blood pressure rise and others small or no rise. These results in the species phylogenetically closest to humans bear directly on causation of human hypertension, particularly in relation to migration of preliterate people, with low Na diet, to a Western urban lifestyle with increased salt intake. The hedonic liking for salt and avid ingestion was apt during human prehistory involving hunter-gatherer-scavenger existence in the interior of continents with a scarcity of salt, but is maladaptive in urban technological life with salt cheap and freely available. PMID- 7489356 TI - Engraftment of gene-modified umbilical cord blood cells in neonates with adenosine deaminase deficiency. AB - Haematopoietic stem cells in umbilical cord blood are an attractive target for gene therapy of inborn errors of metabolism. Three neonates with severe combined immunodeficiency were treated by retroviral-mediated transduction of the CD34+ cells from their umbilical cord blood with a normal human adenosine deaminase complementary DNA followed by autologous transplantation. The continued presence and expression of the introduced gene in leukocytes from bone marrow and peripheral blood for 18 months demonstrates that umbilical cord blood cells may be genetically modified with retroviral vectors and engrafted in neonates for gene therapy. PMID- 7489357 TI - Vascular endothelial growth factor acts as a survival factor for newly formed retinal vessels and has implications for retinopathy of prematurity. AB - Retinopathy of prematurity (ROP) is initiated by hyperoxia-induced obliteration of newly formed blood vessels in the retina of the premature newborn. We propose that vessel regression is a consequence of hyperoxia-induced withdrawal of a critical vascular survival factor. We show that regression of retinal capillaries in neonatal rats exposed to high oxygen, is preceded by a shut-off of vascular endothelial growth factor (VEGF) production by nearby neuroglial cells. Vessel regression occurs via selective apoptosis of endothelial cells. Intraocular injection of VEGF at the onset of experimental hyperoxia prevents apoptotic death of endothelial cells and rescues the retinal vasculature. These findings provide evidence for a specific angiogenic factor acting as a vascular survival factor in vivo. The system also provides a paradigm for vascular remodelling as an adaptive response to an increase in oxygen tension and suggests a novel approach to prevention of ROP. PMID- 7489358 TI - Complete sequencing of the p53 gene provides prognostic information in breast cancer patients, particularly in relation to adjuvant systemic therapy and radiotherapy. AB - The complete coding region of the p53 gene was sequenced from 316 consecutively presented breast cancers, of which 97 were lymph node positive and 206 were node negative. The p53 status was related to prognosis and effect of adjuvant therapy. In all, 69 individual mutations, 29 in node-positive tumours, were demonstrated throughout the whole coding sequence. The mutation sites were partly different for node-positive and node-negative patients. p53 mutations in the evolutionary conserved regions II and V were associated with significantly worse prognosis. Adjuvant systemic therapy, especially with tamoxifen, along with radiotherapy seemed to be of less value to p53 mutation- and lymph node-positive tumours. PMID- 7489359 TI - Individual development and uPA-receptor expression of disseminated tumour cells in bone marrow: a reference to early systemic disease in solid cancer. AB - It is unclear whether disseminated tumour cells detected in bone marrow in early stages of solid cancers indicate a subclinical systemic disease component determining the patient's fate or simply represent mainly irrelevant shed cells. Moreover, characteristics differentiating high and low metastatic potential of disseminated tumour cells are not defined. We performed repeated serial bone marrow biopsies during follow-up in operated gastric cancer patients. Most patients with later tumour relapse revealed either an increase or a constantly high number of tumour cells. In contrast, in patients without recurrence, either clearance of tumour cells or negative or low cell counts were seen. Urokinase plasminogen activator (uPA)-receptor expression on disseminated tumour cells was significantly correlated with increasing tumour cell counts and clinical prognosis. These results demonstrate a systemic component in early solid cancer, indicated by early systemically disseminated tumour cells, which may predict individual disease development. PMID- 7489360 TI - Identification of an endogenous inhibitor of prostatic carcinoma cell growth. AB - The rate of expansion of primary prostatic carcinoma is comparatively slow, with tumours frequently taking years or decades to reach clinically relevant size. We now report the presence of an endogenous inhibitor, derived from aqueous extracts of human prostate tissue, which blocks prostatic carcinoma cell proliferation in vitro and prevents subcutaneous tumour expansion in vivo. Purification and characterization revealed the inhibitor to be spermine, a polyamine known to be locally abundant in the prostate. These results suggest that endogenous polyamine can negatively regulate the growth of prostatic carcinoma cells at their primary site in vivo and may explain the slow rate of primary tumour expansion in the prostate. PMID- 7489361 TI - Discovery of betulinic acid as a selective inhibitor of human melanoma that functions by induction of apoptosis. AB - As a result of bioassay-guided fractionation, betulinic acid, a pentacyclic triterpene, was identified as a melanoma-specific cytotoxic agent. In follow-up studies conducted with athymic mice carrying human melanomas, tumour growth was completely inhibited without toxicity. As judged by a variety of cellular responses, antitumour activity was mediated by the induction of apoptosis. Betulinic acid is inexpensive and available in abundant supply from common natural sources, notably the bark of white birch trees. The compound is currently undergoing preclinical development for the treatment or prevention of malignant melanoma. PMID- 7489362 TI - The p21 cyclin-dependent kinase inhibitor suppresses tumorigenicity in vivo. AB - The p21 gene encodes a cyclin-dependent kinase inhibitor that affects cell-cycle progression, but the potential of this gene product to serve as a tumour suppressor in vivo has not been established. In this report, we show that the growth of malignant cells in vitro and in vivo is inhibited by expression of p21. Expression of p21 resulted in an accumulation of cells in G0/G1, altered morphology, and cell differentiation, but apoptosis was not induced. Introduction of p21 with adenoviral vectors into malignant cells completely suppressed their growth in vivo and also reduced the growth of established pre-existing tumours. Gene transfer of p21 may provide a molecular genetic approach to arresting cancer cell growth by committing malignant cells irreversibly to a pathway of terminal differentiation. PMID- 7489363 TI - Antibacterial activity of lysozyme and lactoferrin is inhibited by binding of advanced glycation-modified proteins to a conserved motif. AB - Why diabetes is associated with abnormally high susceptibility to infection remains unknown, although two major antibacterial proteins, lysozyme and lactoferrin, have now been shown to specifically bind glucose-modified proteins bearing advanced glycation end products (AGEs). Exposure to AGE-modified proteins inhibits the enzymatic and bactericidal activity of lysozyme, and blocks the bacterial agglutination and bacterial killing activities of lactoferrin. Peptide mapping revealed a single AGE binding domain in lysozyme and two AGE binding domains in lactoferrin; each domain contains a 17- to 18- amino acid cysteine bounded loop motif (CX15-16C) that is markedly hydrophilic. Synthetic peptides corresponding to these motifs in lysozyme and lactoferrin exhibited AGE binding activity, and similar domains are also present in other antimicrobial proteins. These results suggest that elevated levels of AGEs in tissues and serum of diabetic patients may inhibit endogenous antibacterial proteins by binding to this conserved AGE-binding cysteine-bounded domain 'ABCD' motif, thereby increasing susceptibility to bacterial infections in the diabetic population. PMID- 7489364 TI - GM1 ganglioside-bound amyloid beta-protein (A beta): a possible form of preamyloid in Alzheimer's disease. AB - The earliest event so far known that occurs in the brain affected with Alzheimer's disease (AD) is the deposition and fibril formation of amyloid beta protein (A beta). A beta is cleaved from a glycosylated membrane protein, called beta-amyloid protein precursor, and normally secreted into the extracellular space. Here we report on the presence of membrane-bound A beta that tightly binds GM1 ganglioside. This suggests that this novel A beta species, rather than secreted A beta, may act as a 'seed' for amyloid and further that intracellular abnormalities in the membrane recycling already exist at the stage of amyloidogenesis. PMID- 7489367 TI - Genetic predisposition to adverse consequences of anti-cholinesterases in 'atypical' BCHE carriers. AB - Normal butyrylcholinesterase (BuChE), but not several of its common genetic variants, serves as a scavenger for certain anti-cholinesterases (anti-ChEs). Consideration of this phenomenon becomes urgent in view of the large-scale prophylactic use of the anti-ChE, pyridostigmine, during the 1991 Persian Gulf War, in anticipation of nerve gas attack and of the anti-ChE, tacrine, for improving residual cholinergic neurotransmission in Alzheimer's disease patients. Adverse symptoms were reported for subjects in both groups, but have not been attributed to specific causes. Here, we report on an Israeli soldier, homozygous for 'atypical' BuChE, who suffered severe symptoms following pyridostigmine prophylaxis during the Persian Gulf War. His serum BuChE and recombinant 'atypical' BuChE were far less sensitive than normal BuChE to inhibition by pyridostigmine and several other carbamate anti-ChEs. Moreover, atypical BuChE demonstrated 1/200th the affinity for tacrine of normal BuChE or the related enzyme acetylcholinesterase (AChE). Genetic differences among BuChE variants may thus explain at least some of the adverse responses to anti-ChE therapies. PMID- 7489365 TI - Tamoxifen elevates transforming growth factor-beta and suppresses diet-induced formation of lipid lesions in mouse aorta. AB - When C57B16 male mice are fed a high-fat diet, they develop significant fatty streak lesions in the aorta. Addition of tamoxifen (TMX) to a high-fat diet, equivalent to a dose of approximately 1 mg TMX per kg body weight per day, suppressed the diet-induced increase in the area of lipid staining in the aortic sinus of the mice by 88% and in the average number of lesions by 86%. The TMX treated mice had 11% +/- 5% less total plasma cholesterol, with most of the reduction in the high density lipoprotein fraction, whereas plasma triglycerides were significantly elevated, and circulating concentrations of 17 beta-estradiol and testosterone were unaffected. Both circulating and aortic concentrations of active and latent transforming growth factor-beta (TGF-beta) were substantially elevated by TMX. The inhibition of lesion formation may be due, at least in part, to cardiovascular protection by TGF-beta. PMID- 7489366 TI - Identification of a novel dual angiotensin II/vasopressin receptor on the basis of molecular recognition theory. AB - The molecular recognition theory suggests that binding sites of interacting proteins, for example, peptide hormone and its receptor binding site, were originally encoded by and evolved from complementary strands of genomic DNA. To test this theory, we screened a rat kidney complementary DNA library twice: first with the angiotensin II (AII) followed by the vasopressin (AVP) antisense oligonucleotide probe, expecting to isolate cDNA clones of the respective receptors. Surprisingly, the identical cDNA clone was isolated twice independently. Structural analysis revealed a single receptor polypeptide with seven predicted transmembrane regions, distinct AII and AVP putative binding domains, a Gs protein-activation motif, and an internalization recognition sequence. Functional analysis revealed specific binding to both AII and AVP as well as AII- and AVP-induced coupling to the adenylate cyclase second messenger system. Site-directed mutagenesis of the predicted AII binding domain obliterates AII binding but preserves AVP binding. This corroborates the dual nature of the receptor and provides direct molecular genetic evidence for the molecular recognition theory. PMID- 7489368 TI - Magnetic resonance mapping demonstrates benefits of VEGF-induced myocardial angiogenesis. AB - Coronary occlusive disease is the leading cause of death in industrial nations and affects one in four adults. Although heart attacks are caused by occlusion of a coronary artery, some patients have occlusions without infarction because they have sufficient collateral vessels providing an alternate pathway for blood supply. Vascular endothelial growth factor (VEGF) is an angiogenic peptide that can stimulate collateral vessel development in the ischaemic myocardium. We used magnetic resonance imaging (MRI) and image processing to identify and quantify non-invasively the benefits related to VEGF infusion on collateral development in the heart. This was accomplished as a placebo-controlled study in the porcine model of chronic ischaemia that most closely mimics the human pathophysiology of progressive coronary occlusion. Image series converted to a space-time map demonstrated that with treatment the ischaemic zone was smaller and the contrast arrival delay was less, which resulted in better ejection fraction and regional wall thickening. These findings demonstrate in a manner applicable to humans, that VEGF improves collateral blood supply, resulting in improved cardiac global and regional function after and in spite of coronary artery occlusion. PMID- 7489369 TI - Microfabricated chemical measurement systems. PMID- 7489370 TI - Rising health-care costs inevitable. PMID- 7489371 TI - Networks for pain. PMID- 7489372 TI - Clinical trials: testing the hypothesis. PMID- 7489373 TI - Rats and hantavirus disease. PMID- 7489374 TI - Treating rheumatoid arthritis with clotrimazole. PMID- 7489375 TI - Heterozygotes for the delta F508 cystic fibrosis allele are not protected against bronchial asthma. PMID- 7489376 TI - Euthanasia: the last word. PMID- 7489377 TI - Viral load: to treat or not to treat? PMID- 7489378 TI - India refining simple HIV test. PMID- 7489379 TI - Confusion reigns over compassionate use of AIDS drug. PMID- 7489380 TI - The hypocrisy of US tobacco policy. PMID- 7489381 TI - HIV: the more things change, the more they stay the same. PMID- 7489382 TI - Dietary salt and blood pressure. PMID- 7489383 TI - MR cardiovascular imaging. PMID- 7489385 TI - The greatest threat of all. PMID- 7489384 TI - Alzheimer's amyloid of another flavour. PMID- 7489386 TI - The molecular recognition theory applied to bispecific antibodies... PMID- 7489387 TI - . . . but not to protein folding? PMID- 7489388 TI - Sterilizing dental equipment. PMID- 7489389 TI - We say 'xeno', you say 'zevos'. PMID- 7489391 TI - Taking tamoxifen to heart. PMID- 7489390 TI - Cholinergic symptoms and Gulf War syndrome. PMID- 7489392 TI - AIDS summit approaches. PMID- 7489394 TI - Terms set for commercializing gene therapy in Japan. PMID- 7489393 TI - Finding new uses for thalidomide. PMID- 7489395 TI - Plague in India. PMID- 7489396 TI - The CD4/CD8 ratio: message in a bottle? PMID- 7489397 TI - Clarifying the causes of Crohn's. PMID- 7489398 TI - Cimetidine and colorectal cancer--old drug, new use? PMID- 7489399 TI - Unrestricted cell cycling and cancer. PMID- 7489400 TI - Computational neuroscience and neurology. PMID- 7489401 TI - Another step towards xenotransplantation. PMID- 7489402 TI - Refining the origins of breast cancer. PMID- 7489403 TI - Assessment of sleepiness in clinical practice. PMID- 7489404 TI - Support for the equivalent light hypothesis for RP. PMID- 7489405 TI - Overexpression of cyclin D mRNA distinguishes invasive and in situ breast carcinomas from non-malignant lesions. AB - The elucidation of molecular alterations that occur during human breast cancer progression may contribute to the development of preventative strategies. Using in situ hybridizations on a cohort of 94 biopsy lesions, quantitatively increased cyclin D mRNA expression levels were observed in only 18% of benign lesions, which confer no or slightly increased breast cancer risk, and 18% of premalignant atypical ductal hyperplasias, which confer a four to fivefold increase in breast cancer risk. The transition to carcinoma was accompanied by frequent cyclin D mRNA overexpression in 76% of low-grade ductal carcinomas in situ, 87% of higher grade comedo ductal carcinomas in situ and 83% of infiltrating ductal breast carcinomas. The data identify a molecular event that may separate benign and premalignant human breast lesions from any form of breast carcinoma. PMID- 7489406 TI - Enzymatic remodelling of the carbohydrate surface of a xenogenic cell substantially reduces human antibody binding and complement-mediated cytolysis. AB - The major obstacle to successful discordant xenotransplantation is the phenomenon of hyperacute rejection (HAR). In the pig-to-primate discordant transplant setting, HAR results from the deposition of high-titre anti-alpha-galactosyl antibodies and complement activation leading to endothelial cell destruction and rapid graft failure. To overcome HAR, we developed an enzymatic carbohydrate remodelling strategy designed to replace expression of the Gal alpha-1,3-Gal xenoepitope on the surface of porcine cells with the non-antigenic universal donor human blood group O antigen, the alpha-1,2-fucosyl lactosamine moiety (H epitope). Xenogenic cells expressing the human alpha-1,2-fucosyltransferase expressed high levels of the H-epitope and significantly reduced Gal alpha-1,3 Gal expression. As a result, these cells were shown to be resistant to human natural antibody binding and complement-mediated cytolysis. PMID- 7489407 TI - Megadose of T cell-depleted bone marrow overcomes MHC barriers in sublethally irradiated mice. AB - Graft-versus-host disease (GVHD) is uniformly lethal in recipients of HLA mismatched marrow. In patients with severe combined immunodeficiency disease, this major obstacle can be overcome by rigorous T-cell depletion before transplantation. In leukaemia patients, however, the benefit of preventing GVHD is offset by graft rejection or graft failure. Very recently, this problem was overcome by supplementing T cell-depleted bone marrow transplants with megadoses of peripheral blood stem cells collected by leukapheresis after mobilization of the donor stem cells with granulocyte colony-stimulating factor (G-CSF). In the present study, we further demonstrate in a mouse model (C57BL/6-->C3H/Hej) that escalation of bone marrow doses by four- to fivefold leads to full donor-type chimerism in sublethally irradiated (6.5 Gy) recipients. Thus, the new source of G-CSF mobilized human haematopoietic stem cells may enable extending the use of mismatched bone marrow transplants to patients with non-malignant diseases for whom supralethal conditioning is not a prerequisite. PMID- 7489408 TI - Kaposi's sarcoma-associated herpesvirus infects endothelial and spindle cells. AB - Kaposi's sarcoma (KS), a vascular tumour that contains characteristic spindle cells forming slit-like spaces, may have an infectious aetiology. Recently, sequences of a new human herpesvirus, KSHV/HHV-8, have been identified in both HIV-associated and classical KS. We sought to identify the target cell of this virus in KS tumour tissue. Using PCR in situ hybridization (PCR-ISH) we show that KSHV/HHV-8 is present in the flat endothelial cells lining vascular spaces of KS lesions as well as in typical KS spindle cells. These findings show that KSHV/HHV 8 is present in the cell types thought to represent neoplastic cells in these lesions. PMID- 7489409 TI - Genetic control of the CD4/CD8 T-cell ratio in humans. AB - We studied the genetic pattern of inheritance of the ratio between circulating CD4+ and CD8+ T lymphocytes in a population of healthy donors. The distribution of the CD4/CD8 ratio in males and females was significantly different and was significantly affected by age. In 46 randomly selected families, the parental CD4/CD8 ratio significantly influenced the ratio in offspring. Complex segregation analysis of the data rejected the non-genetic hypothesis; among the genetic models tested, a major recessive gene with a polygenic component and random environmental effects was the most parsimonious model. These findings indicate that the ratio of CD4+ and CD8+ T cells is genetically controlled in humans. PMID- 7489410 TI - In vivo fate of HIV-1-infected T cells: quantitative analysis of the transition to stable latency. AB - Although it is presumed that the integration of HIV-1 into the genome of infected CD4+ T lymphocytes allows viral persistence, there has been little direct evidence that CD4+ T cells with integrated provirus function as a latent reservoir for HIV-1 in infected individuals. Using resting CD4+ T-cell populations of extremely high purity and a novel assay that selectively and unambiguously detects integrated HIV-1, we show that resting CD4+ T cells harbouring integrated provirus are present in some infected individuals. However, these cells do not accumulate within the circulating pool of resting CD4+ T cells in the early stages of HIV-1 infection and do not accumulate even after prolonged periods in long-term survivors of HIV-1 infection. These results suggest that because of viral cytopathic effects and/or host effector mechanisms, productively infected CD4+ T cells do not generally survive for long enough to revert to a resting memory state in vivo. PMID- 7489411 TI - The Swedish mutation causes early-onset Alzheimer's disease by beta-secretase cleavage within the secretory pathway. AB - Several missense mutations causing early-onset Alzheimer's disease (AD) have been described in the gene coding for the beta-amyloid precursor protein (beta APP). A double mutation found in a Swedish family is located before the amyloid beta peptide (A beta) region of beta APP and results in the increased production and secretion of A beta. Here we show that the increased production of A beta results from a cellular mechanism, which differs substantially from that responsible for the production of A beta from wild-type beta APP. In the latter case, A beta generation requires reinternalization and recycling of beta APP. In the case of the Swedish mutation the N-terminal beta-secretase cleavage of A beta occurs in Golgi-derived vesicles, most likely within secretory vesicles. Therefore, this cleavage occurs in the same compartment as the alpha-secretase cleavage, which normally prevents A beta production, explaining the increased A beta generation by a competition between alpha- and beta-secretase. PMID- 7489412 TI - Bone marrow-derived dendritic cells pulsed with synthetic tumour peptides elicit protective and therapeutic antitumour immunity. AB - Dendritic cells, the most potent 'professional' antigen-presenting cells, hold promise for improving the immunotherapy of cancer. In three different well characterized tumour models, naive mice injected with bone marrow-derived dendritic cells prepulsed with tumour-associated peptides previously characterized as being recognized by class I major histocompatibility complex restricted cytotoxic T lymphocytes, developed a specific T-lymphocyte response and were protected against a subsequent lethal tumour challenge. Moreover, in the C3 sarcoma and the 3LL lung carcinoma murine models, treatment of animals bearing established macroscopic tumours (up to 1 cm2 in size) with tumour peptide-pulsed dendritic cells resulted in sustained tumour regression and tumour-free status in more than 80% of cases. These results support the clinical use of tumour peptide pulsed dendritic cells as components in developing effective cancer vaccines and therapies. PMID- 7489413 TI - Therapeutic gene delivery in human B-lymphoblastoid cells by engineered non transforming infectious Epstein-Barr virus. AB - The B-lymphotrophic human herpes Epstein-Barr virus (EBV) is a 160-kilobase double-stranded DNA episomal virus carried in a persistent asymptomatic state by more than 90% of the worldwide adult population. We engineered a helper-dependent mini-EBV, with the minimal cis-EBV elements for episomal replication, viral amplification and packaging, for use as a gene delivery system. The therapeutic potential of this system was established by stably transducing B-lymphoblastoid cells from a Fanconi anaemia group C (FA-C) patient with a mini-EBV constitutively expressing the normal FACC cDNA and showing in vitro correction of the FA phenotype. In the absence of selective pressure, episomal expression persisted with a half-life of 30 days in actively growing transduced cells, indicating a retention rate of 98% expression per cell doubling. This work demonstrates the generation of an infectious non-transforming viral vector that can potentially deliver large therapeutic genes efficiently and selectively into human B cells. PMID- 7489415 TI - Leptin levels reflect body lipid content in mice: evidence for diet-induced resistance to leptin action. AB - The regulation of body weight and composition involves input from genes and the environment, demonstrated, for example, by the variable susceptibility of inbred strains of mice to obesity when offered a high-fat diet. The identification of the gene responsible for obesity in the ob/ob mouse provides a new approach to defining links between diet and genetics in the regulation of body weight. The ob gene protein product, leptin, is an adipocyte-derived circulating protein. Administration of recombinant leptin reduces food intake and increases energy expenditure in ob/ob mice, suggesting that it signals to the brain the magnitude of fat stores. Information on the regulation of this protein is limited. In several rodent models of obesity including db/db, fa/fa, yellow (Ay/a) VMH lesioned, and those induced by gold thioglucose, monosodium glutamate, and transgenic ablation of brown adipose tissue, leptin mRNA expression and the level of circulating leptin are increased, suggesting resistance to one or more of its actions. We have assessed the impact of increased dietary fat on circulating leptin levels in normal FVB mice and FVB mice with transgene-induced ablation of brown adipose tissue. We find that high-fat diet evokes a sustained increase in circulating leptin in both normal and transgenic mice, with leptin levels accurately reflecting the amount of body lipid across a broad range of body fat. However, despite increased leptin levels, animals fed a high-fat diet became obese without decreasing their caloric intake, suggesting that a high content of dietary fat changes the 'set point' for body weight, at least in part by limiting the action of leptin. PMID- 7489414 TI - Vitamin D receptor genotypes in primary hyperparathyroidism. AB - Vitamin D and parathyroid hormone (PTH) constitute the main regulators of systemic calcium homeostasis. As well as its calcaemic effects, active vitamin D3(1,25(OH)2D3) has a direct regulatory role on parathyroid cells. Active vitamin D3 acts via its receptor (VDR), and binding of the ligand-receptor complex to specific promoter regions of the PTH gene inhibits transcription. Active vitamin D3 constitutes a principal regulator of parathyroid cell growth, and polymorphism in the VDR gene has recently been related to bone mineral density and suggested as predisposing to osteoporosis. Impaired effects of active vitamin D3 may contribute to the relatively enhanced secretion and cell proliferation seen in hyperparathyroidism (HPT). Indeed, VDR dysfunction, of essentially unknown character, has been demonstrated in the pathological parathyroid tissue of primary HPT as well as HPT secondary to uraemia. Consistent with the essential role of active vitamin D3 in parathyroid regulation, the VDR gene polymorphism was studied in 90 postmenopausal women with primary hyperparathyroidism. The VDR genotype bb was found in 60.0% of HPT patients and in 33.3% of the postmenopausal female controls (P < 0.001). As the b allele has been linked to decreased transcriptional activity or messenger RNA stability, reduced VDR expression may impede regulatory actions of vitamin D and may contribute to parathyroid tumorigenesis in these patients. PMID- 7489416 TI - Infection of human tonsil histocultures: a model for HIV pathogenesis. PMID- 7489417 TI - Bone tissue engineering. PMID- 7489418 TI - Caudal analgesia with buprenorphine for postoperative pain relief in children. AB - Caudal buprenorphine was investigated as a postoperative analgesic in a randomized double blind study in thirty children aged 5-12 years undergoing lower abdominal and lower limb surgery. Comparison was made between two groups of patients, one group receiving plain bupivacaine and the other a combination of plain bupivacaine with buprenorphine. Postoperative analgesia was assessed using a linear analogue scale, and by the response to direct questioning of children using an illustration of sequence of faces. Any untoward side effects and the need for additional analgesics were recorded. The degree and duration of analgesia was far superior in the buprenorphine group and there was a highly significant difference in the requirement of postoperative analgesia between the two groups. There were no major adverse side effects and no motor weakness in either groups, however the incidence of nausea and vomiting was higher in the buprenorphine group. It is concluded that a combination of bupivacaine with buprenorphine administered through the caudal epidural space is a safe and reliable means of providing postoperative pain relief in children for up to 24 h. PMID- 7489419 TI - Anaesthetic management of bone marrow transplant recipients less than two years of age. AB - The charts and anaesthetic records of 97 infants less than two years of age who underwent bone marrow transplantation at the University of Minnesota from 1978 1992 were retrospectively reviewed. These infants underwent 564 general anaesthetics. There were 48 perioperative complications, most (39) involving the airway. There were 20 difficult intubations occurring in 13 patients. The causes of the difficult intubations were anatomical abnormalities (12), mucositis (4), pharyngeal oedema (3) and emesis upon induction of anaesthesia (1). Four intraoperative deaths occurred. The deaths were caused by haemorrhage (2), pulmonary embolism (1) and myocardial ischaemia (1). Four patients died within 72 h of surgery; one from cerebral oedema following an intraoperative cardiac arrest, one from fungal septicaemia, one from haemorrhage and one from multiple organ failure following an intracerebral haematoma. Infants undergoing bone marrow transplantation are at high risk for perioperative morbidity and mortality, particularly from complications involving the airway, bleeding or sepsis. PMID- 7489422 TI - New regimen for interpleural block in children. AB - A new regimen for postoperative analgesia after thoracic surgery is proposed. Eight children received an interpleural infusion using bupivacaine 0.1% in a regimen from 0.5 ml.kg-1.h-1 up to 1 ml.kg-1.h-1, for 48 h according to the pain scores. The plasma levels after 24 h and 48 h were measured as well as the pleural level and in two patients the free fraction of plasma bupivacaine and the plasma PPX (a metabolite of bupivacaine) and one patient the orosomucoid (main plasma protein involved in bupivacaine protein binding) were also measured pre and postoperatively. The results shows the safety of such a regimen, for two days of postoperative analgesia. PMID- 7489420 TI - Duration of apnoea in anaesthetized children required for desaturation of haemoglobin to 95%: comparison of three different breathing gases. AB - In this study, we compared three gas compositions to determine if the duration of apnoea for SpO2 to decrease is proportionate to the oxygen fraction of the gas prior to apnoea. Twenty-five patients ASA physical status 1-2 aged two months to 12 years were included in the study. Anaesthesia was induced via a mask with 5% sevoflurane and 66% N2O in oxygen. After paralysis with vecuronium (0.12 mg.kg-1, i.v.) the trachea was intubated and anaesthesia was maintained with sevoflurane and N2O in oxygen. When cardiovascular stability was obtained, the patient was randomly set to breathe one of three gas compositions: 1. oxygen (FiO2 1.0), 2. N2O/O2 (FiO2 0.4), and 3. air/O2 (FiO2 0.4). All three gas compositions included 2-4% of sevoflurane to maintain anaesthesia. After more than eight min of each gas breathing, apnoea was begun by disconnecting the breathing circuit from the tracheal tube. The time from the start of apnoea (SpO2 100%) to SpO2 of 95% (T95) was measured. T95 measured after breathing N2O/O2 and air/O2 were 34.6 +/- 5.7 and 28.8 +/- 4.7% of that measured after oxygen breathing (P < 0.001 vs oxygen breathing, P < 0.001 vs oxygen and N2O/O2 breathing), respectively. Preoxygenation before intubation was validated to delay the haemoglobin desaturation brought about by apnoea. An induction technique using a low FiO2 will allow rapid haemoglobin desaturation. PMID- 7489421 TI - Rapid intravenous administration of ondansetron or metoclopramide is not associated with cardiovascular compromise in children. AB - This double blinded, placebo controlled, randomized, and prospective study investigated the effect of the rapid intravenous administration of ondansetron 0.15 mg.kg-1 or metoclopramide 0.25 mg.kg-1 on the heart rate, haemoglobin saturation, systolic blood pressure, and diastolic blood pressure in 45 ASA PS I II children between two and 16 years of age prior to elective tonsillectomy. The study groups were not significantly different with respect to age, weight, or gender. We were unable to detect a change in heart rate, systolic or diastolic blood pressure, or haemoglobin saturation following the rapid administration of ondansetron or metoclopramide. We conclude intravenous ondansetron or metoclopramide (for the prevention of postoperative vomiting) are not associated with cardiovascular instability when administered rapidly to healthy children prior to elective surgery. PMID- 7489423 TI - Orotracheal intubation through the laryngeal mask airway in paediatric patients with Treacher-Collins syndrome. AB - The laryngeal mask airway (LMA) is useful as an airway intubator (conduit) for an intubating tracheal bougie or fibreoptic bronchoscope, over which a tracheal tube is passed. However, in our paediatric patients with Treacher-Collins syndrome, only the latter technique was successful. This was attributed to the fact that a posteriorly protruded tongue displaced the LMA, made the glottis move considerably anterior and interfered with the attempts to enter the trachea with a bougie. Downward displacement of the epiglottis, which can sometimes impair the intubation technique through the LMA, was not observed in our patients. Partial obstruction of a tracheal tube within the LMA occurred in one of the patients. PMID- 7489424 TI - Larsen's syndrome: anaesthetic implications. Six case reports. AB - Six cases were examined to review the important anaesthetic implications of Larsen's syndrome. Potential problems arising in these patients are highlighted and emphasis placed on cervical spine and airway/respiratory management. PMID- 7489425 TI - Anaesthetic considerations for the patient with cri du chat syndrome. AB - Cri du chat syndrome is an inherited disease affecting multiple organ systems. Most characteristic is the anatomical abnormality of the larynx resulting in a cat-like cry. Issues important in developing an anaesthetic plan include: anatomical abnormalities of the airway, congenital heart disease, hypotonia, mental retardation, and temperature maintenance. We report the case of a 33-month old patient with cri du chat syndrome undergoing patent ductus arteriosus (PDA) ligation and discuss the anaesthetic issues. PMID- 7489426 TI - Accuracy of references in Paediatric Anaesthesia. PMID- 7489427 TI - Tracheal tube occlusion affecting surgical management of gastroschisis. PMID- 7489428 TI - Total intravenous anaesthesia for patients with osteogenesis imperfecta. PMID- 7489429 TI - Surfactant replacement therapy in neonatal respiratory distress syndrome. PMID- 7489430 TI - Cardiorespiratory changes during laparoscopic fundoplication in children. AB - The results of a prospective assessment of cardiorespiratory changes related to anaesthesia and laparoscopic Nissen fundoplication are described in 25 children aged 1.2 to 14.3 years, weighing 9.0 to 64.0 kg. Respiratory disease or oesophagitis were present in 68% cases. During balanced inhalational anaesthesia, hypotension or bradycardia occurred prior to peritoneal insufflation in three cases of reverse Trendelenburg position. During surgery, intra-abdominal pressure was in the 6-10 mmHg range. Transiently, two patients were hypotensive while ten were hypertensive. PETCO2 gradually increased but only two patients required increased minute ventilation. One bronchial intubation episode developed. Airway complications were related to isoflurane administration. Postoperatively, transient hypoxia (25% cases) was observed during the first 3 h. Analgesia duration was in the 40-1440 min range. Hospital stay was 5.6 +/- 1.5 days (mean +/- SD). Laparoscopic paediatric fundoplication is safe when hypovolaemia and postoperative hypoxia are prevented. PMID- 7489431 TI - Anticholinergic prophylaxis does not prevent emesis following strabismus surgery in children. AB - One hundred and twenty-one children were studied in this prospective, randomized double-blind, placebo-controlled comparison of the effectiveness of anticholinergic prophylaxis for the prevention of emetic symptoms following strabismus surgery. The children were allocated to three groups, to receive placebo (n = 40), glycopyrrolate (n = 40) or atropine (n = 41). The incidence of intraoperative oculocardiac reflex (OCR) and of postoperative emetic symptoms for 24 h was recorded. The incidence of OCR was 55% in the placebo group compared with 5% and 2% in the glycopyrrolate and atropine groups respectively (P < 0.05). Thirty percent (12/40) of patients in the placebo group, 25% (10/40) in the glycopyrrolate group, and 22% (9/41) in the atropine group experienced nausea and/or vomiting (difference not significant). It is concluded that prophylactic administration of anticholinergic agents during strabismus surgery in children despite being effective against the occurrence of the oculocardiac reflex, does not reduce the incidence of emetic symptoms. PMID- 7489432 TI - Role of propofol in paediatric anaesthetic practice. PMID- 7489433 TI - The preoperative management of the child with a heart murmur. PMID- 7489434 TI - A survey of interhospital transport of the critically ill child in the United Kingdom. AB - Nineteen paediatric intensive care units were surveyed by questionnaire to provide information on the number of interhospital transfers, the experience of personal accompanying the critically ill child and the equipment available to maintain intensive care during transfer. Replies were received from 17 units. An estimated 800 transfers are performed annually. Three units routinely send intensive care staff to collect patients with an estimated 60% of transfers performed by a variety of staff from referring hospitals. Most respondents believed that existing arrangements for transfer were unsatisfactory, but only four units said that transfer may be prevented or delayed by lack of facilities. We believe that any plan to centralize paediatric intensive care in the UK should also include the means by which to transfer the patient without increasing the risk to the patient. PMID- 7489435 TI - The effect of ketorolac as an adjuvant to local anaesthetic infiltration for analgesia in paediatric umbilical hernia surgery. AB - After umbilical hernia surgery, and wound infiltration with bupivacaine 0.5%, 17 children were given ketorolac 0.5 mg.kg-1, with 18 controls receiving only the wound infiltration. No child experienced severe pain, but moderate pain was noted in patients in both groups. Objective and subjective pain scores were not different statistically at any point up to the morning after surgery. PMID- 7489436 TI - Prevalence of unsuspected myopathy in infants presenting for clubfoot surgery. AB - The objective of this study was an evaluation of the prevalence of myopathies in paediatric patients scheduled for orthopaedic surgery (clubfoot) performed under regional anaesthesia. Seventeen infants scheduled for lower limb orthopaedic surgery were studied to verify coexisting neuromuscular disorders with electromyography and muscle biopsy during surgery. All surgical procedures were performed under caudal block or spinal anaesthesia, associated with light general anaesthesia. No major cardiorespiratory, neurological or malignant hyperthermic complications (muscle rigidity, arrhythmias, hyperpyrexia) were observed. Combined neurological, electromyographic and biopsy studies showed a high rate of myopathic changes (70%). Performance of clubfoot surgery under light general anaesthesia with regional techniques was free from any problems. The high rate of myopathic changes (70%) observed in the muscle biopsies suggests that precautions should be taken with paediatric patients for clubfoot surgery and a regional anaesthesia technique with adequate monitoring may be helpful to prevent possible malignant hyperthermia related problems. PMID- 7489438 TI - Intrathecal morphine (ITM) for postoperative pain control in children: a comparison with nalbuphine patient controlled analgesia (PCA). AB - This is a retrospective study covering the ten-year period 1984-1993. Single shot spinal morphine (ITM) is compared with PCA nalbuphine for postoperative pain relief in children having abdominal or thoracic procedures. The records of 52 patients meeting selection criteria were examined. Nursing and physician notations were reviewed for hourly pain assessments, evidence of associated complications, respiratory depression, nausea and or vomiting, pruritus, and urinary retention. ITM provided significantly better pain relief (2.2 h in pain) during the first 24 h postoperatively than PCA nalbuphine (9.2 h in pain). With the exception of urinary retention which was significantly more frequent following ITM (58.6%) compared to PCA nalbuphine (8.7%), narcotic related complications were not different between the two groups. No difference in duration of hospital stay or ICU stay could be demonstrated. We conclude that ITM provides better pain relief, without more serious complications, than PCA nalbuphine. We recommend it as a safe, effective technique to treat postoperative pain in children following thoracic or upper abdominal procedures. PMID- 7489437 TI - Nicardipine: perioperative applications in children. AB - Nicardipine is the first intravenously administered dihydropyridine calcium channel blocker. Its primary physiological actions include vasodilatation with limited effects on the inotropic and dromotropic function of the myocardium. Several reports have documented its use in adult patients for pharmacological control of blood pressure. We present our experience with the perioperative use of nicardipine in children to treat intraoperative hypertension, as an agent for controlled hypotension during spinal fusion and LeFort I maxillary osteotomies and to treat postoperative hypertension. Dosing regimens and possible applications in paediatric anaesthesia are discussed. PMID- 7489439 TI - Postoperative morphine requirements, nausea and vomiting following anaesthesia for tonsillectomy. Comparison of intravenous morphine and non-opioid analgesic techniques. AB - Nonsteroidal anti-inflammatory drugs (NSAIDs) have been shown to be as effective as opioid analgesia following tonsillectomy in children. Opioids are still frequently used but tonsillectomy is associated with a high incidence of vomiting. This study has attempted to assess postoperative analgesic consumption and nausea and vomiting after general anaesthesia for tonsillectomy using either paracetamol premedication, paracetamol plus a NSAID or intravenous morphine to provide postoperative analgesia. Some children required a rescue dose of morphine in the recovery room, including some who had received intravenous morphine at induction. Least supplementary morphine was required by those who had received paracetamol plus ketorolac. Postoperative nausea and vomiting was significantly less in the two groups which were not given intraoperative morphine. The number of vomiting incidents was also much less. We conclude that the preoperative administration of paracetamol alone provides satisfactory analgesia in many children but that supplementary analgesia is still required for some. PMID- 7489440 TI - Independent right lung high frequency and left lung conventional ventilation in the management of severe air leak during ARDS. AB - Pulmonary air leaks are one of the most common problems in patients with the adult respiratory distress syndrome, ARDS. We report what we believe to be the first case in which unilateral high-frequency ventilation combined with contralateral conventional positive pressure ventilation has been used successfully to manage severe air leak in an infant with ARDS. PMID- 7489442 TI - The laryngeal mask airway for fibreoptic bronchoscopy in children. AB - A size 2 laryngeal mask airway (ID 7 mm) was used for general anaesthesia during fibreoptic bronchoscopy in a six-year-old child. The mask permitted the use of an adult bronchoscope with an external diameter of 5 mm. Throughout bronchoscopy, adequate controlled ventilation could be achieved easily without excessive air leak or airway resistance; the Spo2 was always > 97%, and the end-tidal Pco2 ranged between 3.9-4.5 kPa (30-35 mmHg). PMID- 7489443 TI - Magill's endotracheal catheter device for use during repair of cleft lip and palate. PMID- 7489441 TI - Nitric oxide in the management of persistent pulmonary hypertension of the newborn--an unusual cause of failure. AB - A diagnosis of persistent pulmonary hypertension of the newborn should be reviewed if there is a failure to respond to appropriate treatment. We report a case in which both conventional therapy and inhaled nitric oxide failed to break the cycle of right-to-left shunting. Reassessment revealed a massive pulmonary arteriovenous malformation for which surgery was successfully performed in the neonatal period. PMID- 7489444 TI - Caudal block in a child with osteogenesis imperfecta, type II. PMID- 7489445 TI - Day case survey of postoperative nausea and vomiting. PMID- 7489446 TI - Tracheal tube size and trisomy 21. PMID- 7489447 TI - Bubbles, bubbles, everywhere. PMID- 7489448 TI - Parental presence during induction of anaesthesia. PMID- 7489449 TI - Local anaesthetic agents in infancy. PMID- 7489450 TI - The pharmacokinetics of bupivacaine following interpleural nerve block in infants of very low birthweight. AB - Infants of very low birthweight (VLBW) who underwent thoracotomy were given 2.0 mg.kg-1 of bupivacaine by the intrapleural route, and serial blood levels were taken to determine the pharmacokinetic profile in this group of babies. It was apparent that the half life was longer, clearance lower, and volume of distribution greater than in term infants. Although the drug did not reach toxic levels at this dose, caution should be observed when redosing as the accumulation of the drug may be unpredictable. PMID- 7489451 TI - Femoral and lateral femoral cutaneous nerve block for muscle biopsies in children. AB - Retrospective chart review (1978-1993) of 179 children less than age 18 (10.0 +/- 3.8 SD yrs) undergoing muscle biopsy for determination of susceptibility to malignant hyperthermia provided data. One hundred and forty-six patients received femoral and lateral femoral cutaneous nerve blocks as their primary anaesthetic. We examined age, weight, duration of surgery, time to discharge from hospital, choice and dosage of local anaesthetics, choice and dosage of sedation, postoperative pain medications, and complications. All children receiving this form of anaesthesia remained outpatients. Between 1978 and 1985 procaine (10 mg.kg-1) with hyaluronidase or 2-chloroprocaine (12 mg.kg-1) provided nerve blockade; after 1985, lignocaine (6.8 mg.kg-1), or a combination of lignocaine or mepivacaine and 2-chloroprocaine, were the preferred agents. More recently the combination of 2-chloroprocaine and bupivacaine has been popular. Three patients required admission to the recovery room postoperatively, due to heavy sedation. Forty-three children (29%) received pain medication during recovery. Femoral and lateral femoral cutaneous block anaesthesia with light to moderate sedation is well tolerated in children undergoing anterior thigh procedures. PMID- 7489452 TI - Continuous epidural block for pain control in bladder exstrophy: report of a case and description of technique. PMID- 7489453 TI - Sinus node recovery function is relatively well preserved under halothane anaesthesia in children. AB - The purpose of this study was to evaluate the effect of halothane on sinus node recovery time (SNRT). Nine children with congenital heart disease scheduled for cardiac catherization were studied. After the routine procedure, the sinus node was paced at rates of 120, 150 and 180 beats per minute. SNRTs were measured with control conditions and under 1 MAC halothane anaesthesia. The absolute SNRT was standardized and expressed as '%SNRT' and 'Corrected SNRT (CSNRT)'. CSNRT was slightly prolonged by halothane (P = 0.042), while the %SNRT was not significantly prolonged. We conclude that the recovery function of the sinus node is relatively well preserved under 1 MAC halothane in children. PMID- 7489454 TI - Pharmacokinetics of rectal paracetamol after major surgery in children. AB - Glycogelatin capsular suppositories containing a paracetamol slurry 40 mg.kg-1 were given PR to 20 children (12 months-17 yrs) after major orthopaedic surgery and plasma concentrations of paracetamol measured for up to 18 h. The mean maximum concentration (Cmax) was 0.115 (SD 0.049) mmol.l-1. Peak concentration occurred (Tmax) at 2.3 (SD 1.2) h. Mean concentration was 0.07 (SD 0.03) mmol.l-1 at six h. Apparent paracetamol clearance was 5.8 ml.min-1.kg-1. The plasma concentration of paracetamol associated with analgesic effectiveness in children is unknown, but antipyretic effects are seen in the range 0.066-0.130 mmol.l-1. Paracetamol suppositories 40 mg.kg-1 given perioperatively achieve effective therapeutic antipyretic plasma concentrations within 1-2 h. The timing is coincident with the recovery phase of short duration paediatric surgery. The coefficient of variance of Cmax was 43%. Some individual patients may not achieve a Cmax which is therapeutic. PMID- 7489455 TI - Assessment of the effect of head and neck position on upper airway anatomy in sedated paediatric patients using magnetic resonance imaging. AB - Upper airway patency can be compromised when children (or adults) receive sedative medication. The study examines the effect of two different positioning techniques (use of the 'sniff position pillow' (SPP) and shoulder elevation (SE) on maintenance of upper airway patency using MRI in 21 children sedated with intravenous pentobarbital (5-8 mg.kg-1). Children positioned on the SPP had a significantly greater degree of atlanto-occipital extension (P < 0.05), and a significantly greater nasopharyngeal diameter (P < 0.05) than those with shoulder elevation. The degree of atlanto-occipital extension was not significantly correlated with pharyngeal diameter (R = -0.68). No clinical signs of upper airway obstruction were noted and oxygen desaturation did not occur. Both positioning techniques were consistently associated with upper airway patency under the study conditions described. In obligate nose breathers to whom sedative agents are administered, the SPP is more likely to maintain nasopharyngeal patency than shoulder elevation. PMID- 7489457 TI - Postoperative nausea and vomiting in paediatric surgical inpatients. AB - A prospective study of postoperative nausea and vomiting (PONV) was conducted in 415 children presenting for inpatient surgery. The overall incidence of PONV was 18.1%). The highest incidence was in children undergoing ENT procedures and increased with age. Avoidance of intraoperative opioids and the use of local anaesthesia and/or non-steroidal anti-inflammatory drugs reduced the incidence of nausea and vomiting postoperatively. PMID- 7489456 TI - Propofol reduces the incidence of vomiting after tonsillectomy in children. AB - We compared the effect of a propofol-based anaesthetic to an isoflurane-based anaesthetic on the incidence of postoperative vomiting in children following tonsillectomy. Thirty-nine children were enrolled in the study and randomized to receive one of the proposed anaesthetics. All patients underwent a mask induction with halothane, nitrous oxide, and oxygen. Intravenous access was established and all children received fentanyl (2-4 micrograms.kg-1) i.v., mivacurium (0.3 mg.kg 1) i.v. and acetaminophen (10-15 mg.kg-1) p.r. Following tracheal intubation, patients received either isoflurane (0.8-1.6%) or propofol (120-180 micrograms.kg 1 min-1) i.v. with nitrous oxide 70%/oxygen 30% for maintenance of anaesthesia. Vital signs were maintained within 20% of baseline. All patients were extubated in the operating room. PACU nursing staff recorded episodes of vomiting for 4-6 h prior to discharge. A telephone interview the following day was also used for data recovery. Age, sex, and duration of the procedure were not significantly different between the two study groups. Of 19 patients who received propofol, four vomited (21%); in contrast, of the 20 patients who received isoflurane, 11 vomited (55%). This difference is significant (P = 0.048 two-tailed Fisher's Exact Test). These data suggest that using propofol for anaesthesia can diminish the incidence of vomiting following tonsillectomy. PMID- 7489459 TI - Localized fibromyalgia in a child. AB - A 14-year-old male was investigated because of a limp and a localized sharp pain in the right lumber paravertebral region radiating to the lower abdomen and the medial aspect of the thigh, which started following forced physical activity. With the diagnosis of fibromyalgia the patient received two perifacetal injections of local anaesthetics with steroids followed by transcutaneous electrical nerve stimulation (TENS). Following the injections, pain intensity dropped dramatically, disability was reduced and muscle swelling resolved. The possible association of symptoms to sport activity raises the question of sport induced fibromyalgia, and the excellent response to treatment may suggest a facet joint irritation as possible aetiology. PMID- 7489460 TI - Anaesthesia for a child with centronuclear myopathy. AB - Centronuclear myopathy (CNM) is an inherited condition involving most muscle fibres in all the body mass, first described in 1966, which has a varying spectrum of presentations. Until recently it had not been associated with an increased risk of malignant hyperpyrexia. A seven-year-old male with CNM was admitted to our hospital for elective surgery. High dose propofol anaesthesia was used, supplemented with N2O/O2 from a new anaesthesia machine. The operation was successful with uncomplicated anaesthesia and recovery. PMID- 7489461 TI - Rapid control of severe hypercapnia with high frequency oscillatory ventilation. PMID- 7489458 TI - Anaesthesia in children with viral respiratory tract infections. AB - The effects and consequences of anaesthesia in a child with a respiratory tract infection (RTI) are controversial. There is a high incidence of viral RTI in children presenting for surgery and anaesthesia. The social and economic impact of postponing the procedure is significant; for the child, family and institution. The clinical effects of the common cold are well known, affecting the respiratory tract from the nose down to the small airways and lung parenchyma. The systemic effects of the toxic viraemic phase are also well recognized but not so the potential risk of a viral myocarditis. There is an increased incidence of intra- and postoperative respiratory related complications up to six weeks after a RTI. These include airway obstruction, laryngeal spasm, vagally mediated reflex bronchoconstriction, increased bronchial secretions, desaturation, atelectasis and postoperative respiratory complications. Children with symptoms of a moderate to severe RTI presenting for elective surgery should be postponed for six weeks. Emergency surgery should proceed with a mask anaesthetic for minor surgery or by adopting a modified rapid sequence induction (atropine but no cricoid pressure) to gain rapid control of the airway to avoid laryngeal spasm and vagally mediated reflex bronchoconstriction; IPPV, awake extubation, postoperative monitoring of respiratory function and appropriate analgesia. PMID- 7489462 TI - Paediatric triple lumen catheter. PMID- 7489463 TI - Accidents following extradural analgesia in children. The results of a retrospective study. PMID- 7489464 TI - Paediatric selective bronchial blocker. PMID- 7489465 TI - Absence of corpus callosum and history of slow recovery. PMID- 7489466 TI - What is a paediatric anaesthesiologist? The American perspective. PMID- 7489467 TI - The winds of change. PMID- 7489468 TI - Monitoring of O2 transport and tissue oxygenation in paediatric critical care. PMID- 7489469 TI - Transport for paediatric intensive care. Measuring the performance of a specialist transport service. AB - Fifty children were referred for transport to a paediatric intensive care unit (PICU). Two scoring systems were used for the transfer process. A physiology score derived from the paediatric risk of mortality (PRISM) score was performed at referral, before transfer and on arrival on PICU. An interventions score based on the therapeutic intervention scoring system (TISS) was performed for interventions by the referring staff and by the transport team before and during transfer. Critical events during transport were recorded. Three children died at the referring hospital. Forty-seven were transported by the PICU team. No child died or suffered a major physiological deterioration or equipment related problem in transit. Physiology scores did not deteriorate during transfer. Referral physiology scores did not reliably predict the need for major therapeutic interventions by the transport team before transfer. Critically ill children may be transported safely by a specialist team. PMID- 7489470 TI - Ketamine plus midazolam, a most effective paediatric oral premedicant. AB - Healthy children, 1.5 to seven years old, were divided into three groups of 20 each. Group 1 received midazolam 0.5 mg.kg-1, Group 2, ketamine 6 mg.kg-1 and Group 3 a mixture of midazolam 0.4 mg.kg-1 + ketamine 4 mg.kg-1. Each dose was mixed with atropine 0.02 mg.kg-1 plus an equal volume of cherry syrup and was given orally 20 to 30 min prior to surgery. A grade of 1 (asleep, difficult to arouse), 2 (asleep, easily aroused), 3 (awake, calm), 4 (awake, anxious, occasional cry), or 5 (crying, agitated), was assigned at the time of parental separation and again when mask induction was begun. A grade of 1-3 was considered successful. For parental separation, the mixture of ketamine+midazolam was 100% successful, ketamine 90% and midazolam 75%. Successful mask induction for the mixture of ketamine+midazolam was 85%, midazolam 65% and ketamine 42%. This study indicates that a mixture of ketamine+midazolam is the most effective. PMID- 7489471 TI - Sevoflurane elimination kinetics in children. AB - We compared the rates of elimination of sevoflurane and halothane in 29 children, aged between one and seven years, undergoing ambulatory anaesthesia. Analgesia was provided by fentanyl and muscle relaxation by atracurium. Anaesthesia was maintained by inhalation of one MAC of either sevoflurane or halothane, based on an equipotent concentration of each agent for the age of the child. Following simultaneous discontinuation of N2O and the inhalational agent, the equation describing N2O washout was identical in the presence of halothane and sevoflurane, showing that there was no effect of the volatile agent on the rate of N2O elimination. The elimination of sevoflurane and N2O give similar types of equations. Halothane elimination gives a logarithmic type of equation, showing a slower release, corresponding to residual tissue content. PMID- 7489472 TI - Paediatric ventilatory effects of morphine and buprenorphine revisited. AB - The study describes long term ventilatory effects of 50 or 100 micrograms.kg-1 of morphine or 1.5 or 3.0 micrograms.kg-1 of buprenorphine when given in repeated intravenous (i.v.) doses, in a double blind fashion, to achieve equal levels of analgesia after thoracotomy. The patients were 56 children, six months to six years of age. Ventilatory rate (VR) was measured over the 24 h study period, and arterial carbon dioxide tension (PaCO2) was measured on arrival in the Paediatric Intensive Care Unit (PICU) and at 1, 6, 12 and 18 h. In the buprenorphine groups VRs progressively decreased during the first 2 h and remained significantly lower (P < 0.05) than in the morphine groups for 7 h. For the rest of the study period there were no differences. The PaCO2 values did not differ significantly at any point. For safety, prolonged observation of children is needed after intravenous administration of buprenorphine to ensure the ventilatory rate has stabilized. PMID- 7489473 TI - The safety of continuous pleural lignocaine after thoracotomy in children and adolescents. AB - Several studies have proven pleural bupivacaine effectively provides postthoracotomy analgesia for both children and adults. When 0.25% bupivacaine is administered as a continuous infusion or repeated bolus, serum bupivacaine levels frequently approach the toxic range. The hazards of bupivacaine toxicity are more difficult to monitor, especially in children who may not report symptoms of local anaesthetic toxicity. Because of this concern, we initiated the use of pleural lignocaine to provide postthoracotomy analgesia for paediatric patients. The records of all patients receiving pleural lignocaine from January 1991 to December 1992 were reviewed. A total of 98 pleural catheters were inserted in 96 patients ranging in age from five months to 20 years. Seven patients had lignocaine levels that exceeded 5 micrograms.ml-1 and no patient manifested symptoms of systemic toxicity. This study shows that the administration of pleural lignocaine is a safe method of providing postthoracotomy analgesia. Lignocaine infusions in the dosage range of 20 to 40 micrograms.kg-1.min-1 rarely produce toxic levels, and monitoring of lignocaine levels every 12 h is an effective method of screening for toxicity. PMID- 7489474 TI - Haemodynamic changes in children with portal hypertension during the postoperative period. AB - The cardiovascular changes following portosystemic shunt surgery (PSSS) in 33 children with extrahepatic portal hypertension (EPH) were studied to determine if portosystemic shunt surgery had any influence on the cardiovascular state. Haemodynamic data were obtained using two-dimensional and M-mode echocardiography, pulsed-wave Doppler and direct invasive techniques. Postoperatively all patients developed a hyperdynamic state, associated with an increase in cardiac index (CI), heart rate (HR) and a decrease in systemic vascular resistance index (SVRI). In 16 patients (group I), who preoperatively exhibited a hyperdynamic state accompanied by pulmonary hypertension, postoperative studies found an increase in acceleration time to ejection time ratio (AT/ET) in the pulmonary artery from 0.32 +/- 0.05 (mean +/- SEM) to 0.43 +/- 0.01; P < 0.01. Nine patients (group II) who were normodynamic preoperatively developed pulmonary hypertension and right ventricular (RV) dysfunction postoperatively: decrease in AT/ET from 0.42 +/- 0.02 (mean +/- SEM) to 0.32 +/- 0.01; P < 0.01 and an increase in maximal peak pulmonary artery velocity to acceleration time ratio (Vmax/AT) from 702.56 +/- 69.10 (mean +/- SEM) to 1127 +/ 105.30 cm.sec-2; P < 0.01. In eight patients (group III) who were normodynamic preoperatively and were treated with dobutamine infusions at a rate of 5 micrograms.kg-1.min-1 postoperatively, the CI was found to increase from 5.28 +/- 1.1 (mean +/- SEM) to 7.97 +/- 1.64 L.min-1.m-2; P < 0.01, left ventricular ejection fraction (LVEF) from 69.0 +/- 4.1 (mean +/- SEM) to 81.0 +/- 2.9%; P < 0.01 and AT/ET from 0.37 +/- 0.04 (mean +/- SEM) to 0.44 +/- 0.03; P < 0.01.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7489475 TI - The airway endoscopy mask: useful device for fibreoptic evaluation and intubation of the paediatric airway. AB - A mask is presented which allows the administration of 100% oxygen, inhalational anaesthetics, continuous positive airway pressure and intermittent positive pressure ventilation during diagnostic airway endoscopy and difficult intubation with a fibreoptic bronchoscope in paediatric patients. The mask is particularly useful in small or critically ill patients. It may also have its place in teaching situations. PMID- 7489477 TI - Postoperative airway problems in a child with Crouzon's syndrome. PMID- 7489476 TI - Preoperative investigations. PMID- 7489478 TI - Anaesthetic management of the child with myotonic dystrophy: epidural anaesthesia as an alternative to general anaesthesia. AB - An 11-year-old boy with myotonic dystrophy required anaesthesia for a lower extremity orthopaedic procedure. The management of patients with myotonic dystrophy is discussed including the use of epidural anaesthesia with a continuous chloroprocaine infusion as an alternative to general anaesthesia. PMID- 7489480 TI - Easy blood loss assessment. PMID- 7489479 TI - Potential hazard due to differing markings of paediatric tracheal tubes. PMID- 7489481 TI - Unilateral pulmonary oedema after the modified B-T shunt. PMID- 7489482 TI - Orotracheal intubation through the laryngeal mask airway in paediatric patients with Treacher-Collins syndrome. PMID- 7489483 TI - Purification, cloning, and properties of the tRNA psi 55 synthase from Escherichia coli. AB - tRNA pseudouridine 55 (psi 55) synthase, the enzyme that is specific for the conversion of U55 to psi 55 in the m5U psi CG loop in most tRNAs, has been purified from Escherichia coli and cloned. On SDS gels, a single polypeptide chain with a mass of 39.7 kDa was found. The gene is a previously described open reading frame, p35, located at 68.86 min on the E. coli chromosome between the infB and rpsO genes. The proposed name for this gene is truB. There is very little protein sequence homology between the truB gene product and the hisT (truA) product, which forms psi in the anticodon arm of tRNAs. However, there was high homology with a fragment of a Bacillus subtilis gene that may produce the analogous enzyme in that species. The cloned gene was fused to a 5'-leader coding for a (His)6 tract, and the protein was overexpressed > 400-fold in E. coli. The recombinant protein was purified to homogeneity in one step from a crude cell extract by affinity chromatography using a Ni(2+)-containing matrix. The SDS mass of the recombinant protein was 41.5 kDa, whereas that calculated from the gene was 37.3. The recombinant protein was specific for U55 in tRNA transcripts and reacted neither at other sites for psi in such transcripts nor with transcripts of 16S or 23S ribosomal RNA or subfragments. The enzyme did not require either a renatured RNA structure or Mg2+, and prior formation of m5U was not required. Stoichiometric formation of psi occurred with no requirement for an external source of energy, indicating that psi synthesis is thermodynamically favored. PMID- 7489484 TI - Intrinsic U2AF binding is modulated by exon enhancer signals in parallel with changes in splicing activity. AB - A functional analysis of exon replacement mutations was performed in parallel with RNA-protein binding assays to gain insight into the role of the exon in alternative and simple splicing events. These results show that constitutive exons from unrelated genes contain strong signals that promote splicing in multiple sequence contexts by enhancing 3' splice site activity. A clue to the nature of the relationship between the exon and adjacent 3' splice site is indicated by the binding properties of exon variant RNAs when tested with different biochemical preparations of the essential splicing protein, U2AF. In the context of a complete nuclear extract, U2AF binding to the 3' splice site is stimulated by the presence of an adjacent constitutive exon. In contrast, highly purified HeLa U2AF binds equivalently to the exon variants under conditions in which differential polypyrimidine tract binding is evident. These results provide support for an assisted binding model in which positive-acting signals within exons, exon enhancers, direct the binding of accessory factors, which in turn increase the intrinsic affinity of U2AF for the adjacent 3' splice site. Further support for an assisted binding model is indicated by biochemical complementation of U2AF binding and by the localization of a novel exon enhancer, which, when introduced into a weak exon, stimulates splicing activity in parallel with U2AF binding. Immunoprecipitation analysis identifies the splicing factor, SC35, as a constituent of the exon enhancer binding complex. These results are discussed in the context of current models for functional exon-bridging interactions. PMID- 7489486 TI - Self-assembly of a group I intron active site from its component tertiary structural domains. AB - The catalytic core of Group I self-splicing introns has been proposed to consist of two structural domains, P4-P6 and P3-P9. Each contains helical segments and conserved unpaired nucleotides, and the isolated P4-P6 domain has been shown to have substantial native tertiary structure. The proposed tertiary structure domains of the Tetrahymena intron were synthesized separately and shown to self assemble into a catalytically active complex. Surprisingly, the concentration dependence of these reactions revealed that the domains interact with nanomolar apparent dissociation constants, even though there is no known base pairing between P4-P6 and P3-P9. This suggests that the domains interact through multiple tertiary contacts, the nature of which can now be explored in this system. For example, a circularly permuted version of the P4-P6 domain, which folds similarly to the native P4-P6 molecule, formed a stable but inactive complex. Interestingly, activity was demonstrated with the permuted molecule when nucleotides proposed to form a triple-strand interaction with P4 and P6 were restored as part of the P1-P3 substrate or as part of the P3-P9 RNA. Thus, beyond stabilization of the P4-P6 domain, the triple-strand region may facilitate correct orientation of the RNA domains or participate more directly in catalysis. PMID- 7489485 TI - Proposed nomenclature for the catalytic subunit of the mammalian apolipoprotein B mRNA editing enzyme: APOBEC-1. PMID- 7489488 TI - U6 snRNA function in nuclear pre-mRNA splicing: a phosphorothioate interference analysis of the U6 phosphate backbone. AB - U6 snRNA is essential for and may participate in the catalysis of pre-mRNA splicing. Extensive mutational analyses in several systems have identified nucleotides essential for U6 function in splicing; however, relatively little is known regarding the role of the U6 phosphate backbone. We previously described a mutation in a nematode U6 snRNA that causes it to be used as a splicing substrate within the spliceosome. This unusual reaction has made it possible to apply modification interference analysis to U6 function. Here, we have used phosphorothioate substitution to identify pro-R oxygens throughout the U6 backbone that are necessary for the first and/or second catalytic steps of splicing. Four pro-R oxygens are important for the first step; of these only two appear to be required. One additional pro-R oxygen is uniquely required for the second step. The two pro-R oxygens critical for the first step of splicing are in the helix 1b U2/U6 interaction region and the intramolecular stem-loop of U6, respectively. A comparison of the positions of these two pro-R oxygens with those found to be critical for autocatalytic excision of a group II intron suggests a possible functional similarity between U6 snRNA and domain V of group II introns. PMID- 7489487 TI - Keeping RNA happy. PMID- 7489489 TI - Crosslinking of an iodo-uridine-RNA hairpin to a single site on the human U1A N terminal RNA binding domain. AB - The N-terminal RNA binding domain (RBD) of the human U1A snRNP protein binds tightly and specifically to an RNA hairpin that contains a 10-nucleotide loop. The protein is one of a class of RNA binding proteins that adopts a beta alpha beta beta alpha beta global fold, which in turn forms a four-stranded antiparallel beta-sheet. This sheet forms the primary binding surface for the RNA, as shown by the crosslinking results described here, and in more detail by a recently described co-crystal of this RBD with an RNA hairpin (Oubridge C, et al., 1994, Nature 372:432-438). The RNA hairpin sequence used in the crosslinking experiments, containing 5-iodo-uridine, is a variant of the normal U1 snRNA sequence which is able to form a crosslink with the protein, in contrast to the wild-type sequence, which does not. This single uridine substitution in the 10 nucleotide loop is the site of cross-linking to one tyrosine (Tyr 13) in the beta 1 strand of the U1A N-terminal RBD. This same uridine is also crosslinked to a mutant Tyr 13 Phe RBD, at this Phe 13 substitution. PMID- 7489490 TI - Poly (A) polymerases in the nucleus and cytoplasm of frog oocytes: dynamic changes during oocyte maturation and early development. AB - Poly(A) can be added to mRNAs both in the nucleus and in the cytoplasm. During oocyte maturation and early embryonic development, cytoplasmic polyadenylation of preexisting mRNAs provides a common mechanism of translational control. In this report, to begin to understand the regulation of polyadenylation activities during early development, we analyze poly (A) polymerases (PAPs) in oocytes and early embryos of the frog, Xenopus laevis. We have cloned and sequenced a PAP cDNA that corresponds to a maternal mRNA present in frog oocytes. This PAP is similar in size and sequence to mammalian nuclear PAPs. By immunoblotting using monoclonal antibodies raised against human PAP, we demonstrate that oocytes contain multiple forms of PAP that display different electrophoretic mobilities. The oocyte nucleus contains primarily the slower migrating forms of PAP, whereas the cytoplasm contains primarily the faster migrating species. The nuclear forms of PAP are phosphorylated, accounting for their retarded mobility. During oocyte maturation and early postfertilization development, preexisting PAPs undergo regulated phosphorylation and dephosphorylation events. Using the cloned PAP cDNA, we demonstrate that the complex changes in PAP forms seen during oocyte maturation may be due to modifications of a single polypeptide. These results demonstrate that the oocyte contains a cytoplasmic polymerase closely related to the nuclear enzyme and suggest models for how its activity may be regulated during early development. PMID- 7489491 TI - Interactions between the double-stranded RNA binding motif and RNA: definition of the binding site for the interferon-induced protein kinase DAI (PKR) on adenovirus VA RNA. AB - The protein kinase DAI, the double-stranded RNA activated inhibitor of translation (also known as PKR), regulates cell growth, virus infection, and other processes. DAI represents a class of proteins containing a recently recognized RNA binding motif, the dsRBM, but little is known about the contacts between these proteins and their RNA ligands. In adenovirus-infected cells, DAI activation is prevented by VA RNAI, a highly structured RNA that binds to the kinase. VA RNA contains three chief structural features: a terminal stem, an apical stem-loop, and a complex central domain. We used enzymatic and chemical footprinting to identify the interactions between DAI and VA RNAI. DAI protects the proximal part of the apical stem structure, an adjacent region in the central domain, and a region surrounding a conserved stem in the central domain from nuclease attack. During binding the RNA undergoes a conformational change that is mainly restricted to the central domain. A similar change is induced by magnesium ions alone. Footprinting and interference binding assays using base-specific chemical probes suggest that the protein does not make major contacts with RNA bases. On the other hand, footprinting with probes specific for the RNA backbone shows that DAI engages in a strong interaction with the minor groove of the apical stem and a weaker interaction in the central domain. A truncated form of DAI, p20, containing only the RNA binding domain, gives a similar protection pattern in the apical stem but protects the central domain less effectively. We conclude that the RNA binding domain of DAI interacts directly with the apical stem and central domain of VA RNA, and that other regions of the protein contribute to interactions with the central domain. PMID- 7489492 TI - Translational control of maturation-protein synthesis in phage MS2: a role for the kinetics of RNA folding? AB - The gene for the maturation (A) protein of the single-stranded RNA coliphage MS2 is preceded by an untranslated leader of 130 nt. Secondary structure of the leader was deduced by phylogenetic comparison and by probing with enzymes and chemicals. The RNA folds into a cloverleaf, i.e., three stem-loop structures enclosed by a long-distance interaction (LDI). This LDI is essential for translational control. Its 3'moiety contains the Shine-Dalgarno region of the A protein gene, whereas its complement is located 80 nt upstream, i.e., about 30 nt from the 5'-terminus of the RNA chain. Mutational analysis shows that this base pairing represses expression of the A-protein gene. We present a model in which translational starts can only take place on nonequilibrated RNA, in which base pairing between the complementary regions has not yet taken place. We suggest that this pairing is kinetically delayed by the intervening sequence, which contains the three hairpins of the cloverleaf. The model is mainly based on the observation that reducing the length of the intervening sequence reduces expression, whereas increasing the length has the opposite effect. In addition, further stabilization of the LDI by a stronger base pair does not lead to a decrease in A-protein synthesis. Such a decrease is predicted to occur if translation would be controlled by the equilibrium structure of the leader RNA. These and other observations fit a kinetic model of translational control by RNA folding. PMID- 7489493 TI - A mutation at the universally conserved position 529 in Escherichia coli 16S rRNA creates a functional but highly error prone ribosome. AB - A base substitution of G to U was constructed at position 529 in Escherichia coli 16S rRNA. The U529 mutant ribosomes were functional and present on polysomes but were highly error prone and caused a progressive loss of cell viability. They displayed elevated levels of readthrough of stop codons and frameshifting, and an increase in thermal sensitivity of beta-galactosidase, suggestive of missense errors. These results demonstrate that the university conserved G529 is involved in tRNA selection at the A site during protein synthesis. PMID- 7489494 TI - Inhibition of the hammerhead ribozyme by neomycin. AB - A series of antibiotics was tested for stimulation or inhibition of the hammerhead ribozyme cleavage reaction. Neomycin was found to be a potent inhibitor of the reaction with a Kl of 13.5 microM. Two hammerheads with well characterized kinetics were used to determine which steps in the reaction mechanism were inhibited by neomycin. The data suggest that neomycin interacts preferentially with the enzyme-substrate complex and that this interaction leads to a reduction in the cleavage rate by stabilizing the ground state of the complex and destabilizing the transition state of the cleavage step. A comparison of neomycin with other aminoglycosides and inhibitors of hammerhead cleavage implies that the ammonium ions of neomycin are important for the antibiotic hammerhead interaction. PMID- 7489495 TI - Regulation of the hepatitis delta virus ribozymes: to cleave or not to cleave? PMID- 7489496 TI - Regulation of alternative 3' splice site selection by constitutive splicing factors. AB - We have devised an in vitro splicing assay in which the mutually exclusive exons 2 and 3 of alpha-tropomyosin act as competing 3' splice sites for joining to exon 1. Splicing in normal HeLa cell nuclear extracts results in almost exclusive joining of exons 1 and 3. Splicing in decreased nuclear extract concentrations and decreased ionic strength results in increased 1-2 splicing. We have used this assay to determine the role of three constitutive pre-mRNA splicing factors on alternative 3' splice site selection. Polypyrimidine tract binding protein (PTB) was found to inhibit the splicing of introns containing a strong binding site for this factor. However, the inhibitory effect of PTB could be partially reversed if pre-mRNAs were preincubated with U2 auxiliary factor (U2AF) prior to splicing in PTB-supplemented extracts. For alpha-tropomyosin, regulation of splicing by PTB and U2AF primarily affected the joining of exons 1-3 with no dramatic increases in 1-2 splicing being detected. Preincubation of pre-mRNAs with SR proteins led to small increases in 1-2 splicing. However, if pre-mRNAs were preincubated with SR proteins followed by splicing in PTB-supplemented extracts, there was a nearly complete reversal of the normal 1-2 to 1-3 splicing ratios. Thus, multiple pairwise, and sometimes antagonizing, interactions between constitutive pre-mRNA splicing factors and the pre-mRNA can regulate 3' splice site selection. PMID- 7489497 TI - The non-RNase H domain of Saccharomyces cerevisiae RNase H1 binds double-stranded RNA: magnesium modulates the switch between double-stranded RNA binding and RNase H activity. AB - Eukaryotic ribonucleases H of known sequence are composed of an RNase H domain similar in size and sequence to that of Escherichia coli RNase HI and additional domains of unknown function. The RNase H1 of Saccharomyces cerevisiae has such an RNase H domain at its C-terminus. Here we show that the N-terminal non-RNase H portion of the yeast RNase H1 binds tightly to double-stranded RNA (dsRNA) and RNA-DNA hybrids even in the absence of the RNase H domain. Two copies of a sequence with limited similarity to the dsRNA-binding motif are present in this N terminus. When the first of these sequences is altered, the protein no longer binds tightly to dsRNA and exhibits an increase in RNase H activity. Unlike other dsRNA-binding proteins, increasing the Mg2+ concentration from 0.5 mM to 5 mM inhibits binding of RNase H1 to dsRNA; yet a protein missing the RNase H domain binds strongly to dsRNA even at the higher Mg2+ concentration. These results suggest that binding to dsRNA and RNase H activity are mutually exclusive, and the Mg2+ concentration is critical for switching between the activities. Changes in the Mg2+ concentration or proteolytic severing of the dsRNA-binding domain could alter the activity or location of the RNase H and may govern access of the enzyme to the substrate. Sequences similar to the dsRNA-binding motif are present in other eukaryotic RNases H and the transactivating protein of cauliflower mosaic virus, suggesting that these proteins may also bind to dsRNA. PMID- 7489498 TI - Mammalian splicing factor SF3a120 represents a new member of the SURP family of proteins and is homologous to the essential splicing factor PRP21p of Saccharomyces cerevisiae. AB - Mammalian splicing factor SF3a consists of three subunits of 60, 66, and 120 kDa and functions early during pre-mRNA splicing by converting the U2 snRNP into its active form. A cDNA encoding the 120-kDa subunit of SF3a has been cloned. The SF3a120 gene was localized to human chromosome 22, and three mRNAs of 3.2, 3.8, and 5.7 kb are ubiquitously expressed. The N-terminal half of the deduced SF3a120 amino acid sequence contains a tandemly repeated motif (the SURP module) that has recently been identified in the essential splicing factor PRP21p of Saccharomyces cerevisiae, the Drosophila alternative splicing regulator suppressor-of-white apricot, and four proteins from nematodes and mammals; the C-terminal half is organized into a proline-rich region and a ubiquitin-like domain. The spacing between the SURP modules and the protein's essential function in constitutive splicing identify SF3a120 as the mammalian homologue of yeast PRP21p. Binding studies with truncated derivatives of SF3a120 revealed that the SURP domains function in binding to SF3a60, whereas a region of 130 amino acids C-terminal to these domains is essential for contacts with SF3a66. PMID- 7489499 TI - Nuclear retention of RNA as a mechanism for localization. AB - Two mutant RNAs, one derived from tRNA(imet), the second from U1 snRNA, that are defective in export from the nucleus to the cytoplasm have been studied. In both cases, the RNAs are shown to be transport competent but prevented from leaving the nucleus by interaction with saturable binding sites. This contradicts previous hypotheses to explain the behavior of the tRNA mutant, and highlights a general problem in using mutant RNAs to study nuclear export. In the case of these mutants, it is argued that nuclear retention is likely to be artifactual. However, the additional example of U6 snRNA is described. In this case, nuclear retention appears to be a physiological mechanism by which intranuclear localization is achieved. Evidence that the site of interaction with the La protein in U6 snRNA is important for its nuclear retention is presented. PMID- 7489500 TI - In vivo facilitation of Tetrahymena group I intron splicing in Escherichia coli pre-ribosomal RNA. AB - The observation that the large ribosomal RNA intron of Tetrahymena is spliced 20 50-fold more rapidly in vivo than in vitro (Brehm SL, Cech TR, 1983, Biochemistry 22:2390-2397; Bass BL, Cech TR, 1984, Nature 308:820-826) suggests facilitation of RNA folding in vivo. To determine whether a specific group I splicing factor is required in Tetrahymena, the intron was inserted into the analogous position of the Escherichia coli 23S rRNA. We report that the intron is rapidly excised from pre-rRNA in bacteria and that the magnitude of the in vivo rate enhancement is similar to that in Tetrahymena. These results demonstrate that a species specific protein is not required. Instead, a common mechanism of assisting RNA folding is sufficient to accelerate the removal of self-splicing introns from ribosomal RNA. PMID- 7489501 TI - Caenorhabditis elegans embryos contain only one major species of Ro RNP. AB - In virtually all vertebrate cells, Ro RNPs consist of the 60-kDa Ro autoantigen bound to one of several small cytoplasmic RNA molecules known as Y RNAs. Because the 60-kDa Ro autoantigen is also found complexed with defective precursors of 5S rRNA in Xenopus oocytes, we have proposed that this protein functions in a quality control, or discard pathway, for 5S RNA biosynthesis (O'Brien CA, Wolin SL, 1994, Genes & Dev 8:2891-2903). The role of the Y RNAs in this pathway is unknown. To begin a genetic analysis of Ro RNP function, we have characterized these particles in the nematode Caenorhabditis elegans. The C. elegans Ro protein is 12 kDa larger than the vertebrate protein; the larger size is due in part to an N-terminal extension and to two insertions in the RNA recognition motif. In contrast to all previously described vertebrate species, the Ro protein appears bound to a single Y RNA in C. elegans. Similar to vertebrate Y RNAs, the C. elegans Y RNA can be folded to form a pyrimidine-rich internal loop and a long stem in which the 5' and 3' ends are base paired. Within the stem is a conserved bulged helix that is proposed to be the binding site of the Ro protein. Interestingly, although the human protein can bind the nematode Y RNA, the C. elegans protein does not bind human Y RNAs. This is the first description of Ro RNPs in an invertebrate species. PMID- 7489502 TI - The influenza virus NS1 protein binds to a specific region in human U6 snRNA and inhibits U6-U2 and U6-U4 snRNA interactions during splicing. AB - The influenza virus NS1 protein is a unique posttranscriptional regulator that has two activities: inhibition of the nuclear export of poly A-containing mRNAs and inhibition of pre-mRNA splicing. Here we demonstrate that this protein binds to a specific region in one of the human spliceosomal snRNAs, U6 snRNA. Using U6 deletion mutations, we show that the binding of the NS1 protein requires both chains of a stem-bulge structure encompassing nucleotides 27-46 and nucleotides 83-101 of human U6 snRNA. A chemical modification/interference assay indicated that the primary binding site is centered around a purine-containing bulge in this stem-bulge structure. These results provide strong evidence that this postulated secondary structure in U6 snRNA actually exists. The NS1 protein also binds to a model U6-U4 snRNA complex, suggesting that the U6 stem-bulge comprising the NS1 protein binding site is also present in natural U6-U4 snRNA complexes. The U6 stem-bulge includes the U6 sequence that forms helix II with U2 snRNA during splicing, an interaction that is essential for mammalian splicing. We demonstrate that the NS1 protein blocks formation of the U6-U2 helix II both in a model system and during in vitro splicing. In addition, we show that the NS1 protein inhibits formation of U6-U4 snRNA complexes during in vitro splicing, presumably because the binding site of the NS1 protein includes the 3'-terminal region of U6 snRNA that has been shown to be important for the formation of U6-U4 complexes. We postulate that the inhibition of U6-U2 and U6-U4 snRNA complex formation is largely responsible for the inhibition of pre-mRNA splicing by the NS1 protein. PMID- 7489503 TI - Dissecting protein:protein interactions between transcription factors with an RNA aptamer. AB - Nucleic acid aptamers isolated from random sequence pools have generally proven useful at inhibiting the interactions of nucleic acid binding proteins with their cognate nucleic acids. In order to develop reagents that could also be used to study protein:protein interactions, we have used in vitro selection to search for RNA aptamers that could interact with the transactivating protein Tax from human T-cell leukemia virus. Tax does not normally bind to nucleic acids, but instead stimulates transcription by interacting with a variety of cellular transcription factors, including the cyclic AMP-response element binding protein (CREB), NF kappa B, and the serum response factor (SRF). Starting from a pool of greater than 10(13) different RNAs with a core of 120 random sequence positions, RNAs were selected for their ability to be co-retained on nitrocellulose filters with Tax. After five cycles of selection and amplification, a single nucleic acid species remained. This aptamer was found to bind Tax with high affinity and specificity, and could disrupt complex formation between Tax and NF-kappa B, but not with SRF. The differential effects of our aptamer probe on protein:protein interactions suggest a model for how the transcription factor binding sites on the surface of the Tax protein are organized. This model is consistent with data from a variety of other studies. PMID- 7489504 TI - Processing of the precursor to the catalytic RNA subunit of RNase P from Escherichia coli. AB - M1 RNA, the catalytic subunit of RNase P from Escherichia coli, is transcribed in vivo as a precursor with extra nucleotides at the 3' end. Although it was suggested previously that RNase E is not responsible for the 3' processing of M1 RNA, we show that RNase E is the enzyme responsible for this reaction. At nonpermissive temperatures, the 3' processing of M1 RNA is abolished in a temperature-sensitive strain of E. coli that harbors a mutation in the gene for RNase E. Enhanced processing of M1 RNA is correlated with the overproduction of RNase E in vivo and processing is also correlated with the activity of this enzyme during the course of its purification. The biosynthesis of mature M1 RNA can proceed from transcripts that are produced under the control of a proximal promoter, as well as from a distal, upstream promoter. Transcription from the distal promoter results in a polycistronic transcript that includes four open reading frames and the transcript of rnpB, the gene coding for M1 RNA. The enzymatic activity that removes the 5' nucleotides from the precursor to M1 RNA is not due to RNase E, RNase P, or RNase III alone. PMID- 7489505 TI - Overexpression of the SR proteins ASF/SF2 and SC35 influences alternative splicing in vivo in diverse ways. AB - The SR proteins are a family of essential splicing factors highly conserved throughout metazoa. Here we examine the effects of two prototypical SR proteins, ASF/SF2 and SC35, when overexpressed by transfection in cultured cells together with plasmids encoding alternatively spliced model transcripts. As expected from past work, both proteins were found to affect alternative splicing, but differences as well as similarities in their behavior were observed. With adenovirus E1a pre-mRNA, ASF/SF2 caused shifts in alternative splicing similar to those observed previously, and the effects of mutations, in the protein and the pre-mRNA, were largely consistent with in vitro results. For example, the C terminal RS domain was not required to alter splice site selection. SC35 overexpression also altered E1a splicing, but the pattern was distinct from that detected with ASF/SF2, indicating that the two proteins can function differently in vivo. Unexpectedly, with SV40 early pre-mRNA, overexpression of either protein resulted in a marked inhibition of splicing, with the downstream small t 5' splice site more sensitive than the upstream large T 5' splice site. This is essentially the opposite of what has been observed when the concentration of these proteins is increased in vitro. The RS domain was necessary but not sufficient for this effect. Finally, overexpression of SC35, but not ASF/SF2, resulted in substantial accumulation of the unspliced SV40 pre-mRNA, which was efficiently transported to the cytoplasm. This finding suggests that SC35 may play an unanticipated role in mRNA stability and/or transport. PMID- 7489506 TI - On the road to a DNA-protein world. PMID- 7489508 TI - Functional oligomerization of poliovirus RNA-dependent RNA polymerase. AB - Using a hairpin primer/template RNA derived from sequences present at the 3' end of the poliovirus genome, we investigated the RNA-binding and elongation activities of highly purified poliovirus 3D polymerase. We found that surprisingly high polymerase concentrations were required for efficient template utilization. Binding of template RNAs appeared to be the primary determinant of efficient utilization because binding and elongation activities correlated closely. Using a three-filter binding assay, polymerase binding to RNA was found to be highly cooperative with respect to polymerase concentration. At pH 5.5, where binding was most cooperative, a Hill coefficient of 5 was obtained, indicating that several polymerase molecules interact to retain the 110-nt RNA in a filter-bound complex. Chemical crosslinking with glutaraldehyde demonstrated physical polymerase-polymerase interactions, supporting the cooperative binding data. We propose a model in which poliovirus 3D polymerase functions both as a catalytic polymerase and as a cooperative single-stranded RNA-binding protein during RNA-dependent RNA synthesis. PMID- 7489510 TI - Mutation analyses of molecularly cloned satellite tobacco mosaic virus during serial passage in plants: evidence for hotspots of genetic change. AB - The high level of genetic diversity and rapid evolution of viral RNA genomes are well documented, but few studies have characterized the rate and nature of ongoing genetic change over time under controlled experimental conditions, especially in plant hosts. The RNA genome of satellite tobacco mosaic virus (STMV) was used as an effective model for such studies because of advantageous features of its genome structure and because the extant genetic heterogeneity of STMV has been characterized previously. In the present study, the process of genetic change over time was studied by monitoring multiple serial passage lines of STMV populations for changes in their consensus sequences. A total of 42 passage lines were initiated by inoculation of tobacco plants with a helper tobamovirus and one of four STMV RNA inocula that were transcribed from full length infectious STMV clones or extracted from purified STMV type strain virions. Ten serial passages were carried out for each line and the consensus genotypes of progeny STMV populations were assessed for genetic change by RNase protection analyses of the entire 1,059-nt STMV genome. Three different types of genetic change were observed, including the fixation of novel mutations in 9 of 42 lines, mutation at the major heterogeneity site near nt 751 in 5 of the 19 lines inoculated with a single genotype, and selection of a single major genotype in 6 of the 23 lines inoculated with mixed genotypes. Sequence analyses showed that the majority of mutations were single base substitutions. The distribution of mutation sites included three clusters in which mutations occurred at or very near the same site, suggesting hot spots of genetic change in the STMV genome. The diversity of genetic changes in sibling lines is clear evidence for the important role of chance and random sampling events in the process of genetic diversification of STMV virus populations. PMID- 7489507 TI - When cells stop making sense: effects of nonsense codons on RNA metabolism in vertebrate cells. AB - It appears that no organism is immune to the effects of nonsense codons on mRNA abundance. The study of how nonsense codons alter RNA metabolism is still at an early stage, and our current understanding derives more from incidental vignettes than from experimental undertakings that address molecular mechanisms. Challenges for the future include identifying the gene products and RNA sequences that function in nonsense mediated RNA loss, resolving the cause and consequences of there apparently being more than one cellular site and mechanism for nonsense mediated RNA loss, and understanding how these sites and mechanisms are related to both constitutive and specialized pathways of pre-mRNA processing and mRNA decay. PMID- 7489509 TI - Reverse splicing of the Tetrahymena IVS: evidence for multiple reaction sites in the 23S rRNA. AB - Group I introns in rRNA genes are clustered in highly conserved regions that include tRNA and mRNA binding sites. This pattern is consistent with insertion of group I introns by direct interaction with exposed regions of rRNA. Integration of the Tetrahymena group I intron (or intervening sequence, IVS) into large subunit rRNA via reverse splicing was investigated using E. coli 23S rRNA as a model substrate. The results show that sequences homologous to the splice junction in Tetrahymena are the preferred site of integration, but that many other sequences in the 23S rRNA provide secondary targets. Like the original splice junction, many new reaction sites are in regions of stable secondary structure. Reaction at the natural splice junction is observed in 50S subunits and to a lesser extent in 70S ribosomes. These results support the feasibility of intron transposition to new sites in rRNA genes via reverse splicing. PMID- 7489511 TI - The conformation of 23S rRNA nucleotide A2058 determines its recognition by the ErmE methyltransferase. AB - The ErmE methyltransferase confers resistance to MLS antibiotics by specifically dimethylating adenine 2058 (A2058, Escherichia coli numbering) in bacterial 23S rRNA. To define nucleotides in the rRNA that are part of the motif recognized by ErmE, we investigated both in vivo and in vitro the effects of mutations around position A2058 on methylation. Mutagenizing A2058 (to G or U) completely abolishes methylation of 23S rRNA by ErmE. No methylation occurred at other sites in the rRNA, demonstrating the fidelity of ErmE for A2058. Breaking the neighboring G2057-C2611 Watson-Crick base pair by introducing either an A2057 or a U2611 mutation, greatly reduces the rate of methylation at A2058. Methylation remains impaired after these mutations have been combined to create a new A2057 U2611 Watson-Crick base interaction. The conformation of this region in 23S rRNA was probed with chemical reagents and it was shown that the A2057 and U2611 mutations alone and in combination alter the reactivity of A2058 and adjacent bases. However, mutagenizing position G-->A2032 in an adjacent loop, which has been implicated to interact with A2058, alters neither the ErmE methylation at A2058 nor the accessibility of this region to the chemical reagents. The data indicate that a less-exposed conformation at A2058 leads to reduction in methylation by ErmE. Nucleotide G2057 and its interaction with C2611 maintain the conformation at A2058, and are thus important in forming the structural motif that is recognized by the ErmE methyltransferase. PMID- 7489512 TI - The large genome segment of dsRNA bacteriophage phi6 is the key regulator in the in vitro minus and plus strand synthesis. AB - Bacteriophage phi6 is a double-stranded RNA (dsRNA) virus that has a genome composed of three linear dsRNA segments (L, M, S). These are encapsidated into a dodecahedral procapsid particle consisting of proteins P1, P2, P4, and P7. Empty preformed procapsids are able to package the plus-sense single-stranded RNA (ssRNA) of each genome segment, to synthesize the corresponding minus strands ("replication") to form dsRNA segments, and to continue to the plus strand synthesis ("transcription") in which the dsRNA segments are used as templates in production of plus-sense ssRNA. In this study, we have investigated the requirements for the switch-on of minus and plus strand syntheses. We show that there exists an inverse relationship between regulation of the ssRNA packaging and minus strand synthesis. The packaging of single-stranded l, which has previously been shown to be packaged as the last, is the necessary signal for the onset of the minus strand synthesis. The absolute requirement for plus strand synthesis is minus strand synthesis of l, but in addition, the minus strand synthesis of m and the packaging of s segment are needed for efficient plus strand synthesis. Furthermore, the second nucleotide at the 5'-end of each segment regulates the extent of the transcription. PMID- 7489513 TI - Structure and function of the selenium translation element in the 3'-untranslated region of human cellular glutathione peroxidase mRNA. AB - In eukaryotes, incorporation of selenocysteine into the polypeptide chain at a UGA codon requires a unique sequence motif, or "selenium translation element" (STE), located in the 3'-untranslated region of the mRNA. The present study examines structure-function relationships of conserved sequence elements and of the putative stem-loop secondary structure in the STE of human GPX1 mRNA, which encodes the important antioxidant enzyme cellular glutathione peroxidase (EC 1.11.1.9). Deletion of the basal stem, upper stem, or apical loop of the stem loop structure eliminated the ability of the STE to direct selenocysteine incorporation at the UGA codon of an epitope-tagged GPX1 reporter construct transfected into COS1 cells. However, mutations that change the primary nucleotide sequence of nonconserved portions of the stem-loop, but preserve its overall secondary structure, by inversion of apical loop sequences or exchange of 5' and 3' sides of stem segments, had little or no effect on selenocysteine incorporation. Effects of single- and double-nucleotide substitutions in three short, highly conserved elements in the GPX1 STE depended in large part on their computer-predicted perturbation of the stem-loop and its midstem bulge. Only in the conserved "AAA" apical loop sequence did mutations show major effects on function without predicted changes in secondary structure. Our results demonstrate the critical role of the three short, highly conserved sequences. However, outside of these elements, the function of the human GPX1 STE appears to depend strongly on the stem-loop secondary structure. PMID- 7489514 TI - An RNA pseudoknot is an essential structural element of the internal ribosome entry site located within the hepatitis C virus 5' noncoding region. AB - Translation of the human hepatitis C virus (HCV) RNA genome occurs by a mechanism known as "internal ribosome entry." This unusual strategy of translation is employed by naturally uncapped picornaviral genomic RNAs and several cellular mRNAs. A common feature of these RNAs is a relatively long 5' noncoding region (NCR) that folds into a complex secondary structure harboring an internal ribosome entry site (IRES). Evidence derived from the use of dicistronic expression systems, combined with an extensive mutational analysis, demonstrated the presence of an IRES within the HCV 5'NCR. The results of our continued mutational analysis to map the critical structural elements of the HCV IRES has led to the identification of a pseudoknot structure upstream of the initiator AUG. The evidence presented in this study is based upon the mutational analysis of the putative pseudoknot structure. This is further substantiated by biochemical and enzymatic probing of the wild-type and mutant 5'NCR. Further, the thermodynamic calculations, based upon a modified RNAKNOT program, are consistent with the presence of a pseudoknot structure located upstream of the initiator AUG. Maintenance of this structural element is critical for internal initiation of translation. The pseudoknot structure in the 5'NCR represents a highly conserved feature of all HCV subtypes and members of the pestivirus family, including hog cholera virus and bovine viral diarrhea virus. PMID- 7489515 TI - Selection of an RNA domain that binds Zn2+. AB - We have selected an RNA that depends on zinc for affinity to a column, starting from a pool of ribooligonucleotides with 50 randomized positions. This RNA's chemical sensitivities, calculated folding thermodynamics, and activity when fragmented suggest that an ion binding site lies within a complex 21-nt hairpin loop, near the junction with an imperfect helical stem. This RNA site has an unselected selectivity among divalents, preferring nickel, cobalt, and cadmium to calcium, magnesium, and manganese, as expected for a simple site of chelation. A moderate zinc-dependent change in loop structure accompanies divalent binding and can be detected by chemical probing and zinc-dependent UV-induced crosslinking. The latter also demonstrates the apposition of loop sequences to make a structure that may be related to the E-loop motif found in a number of other RNA molecules; the E-loop motif, accordingly, may be a divalent site. PMID- 7489516 TI - A comparison of thermodynamic foldings with comparatively derived structures of 16S and 16S-like rRNAs. AB - To increase our understanding of the dynamics and complexities of the RNA folding process, and therewith to improve our ability to predict RNA secondary structure by computational means, we have examined the foldings of a large number of phylogenetically and structurally diverse 16S and 16S-like rRNAs and compared these results with their comparatively derived secondary structures. Our initial goals are to establish the range of prediction success for this class of rRNAs, and to begin comparing and contrasting the foldings of these RNAs. We focus here on structural features that are predicted with confidence as well as those that are poorly predicted. Whereas the large set of Archaeal and (eu)Bacterial 16S rRNAs all fold well (69% and 55% respectively), some as high as 80%, many Eucarya and mitochondrial 16S rRNAs are poorly predicted (approximately 30%), with a few of these predicted as low as 10-20%. In general, base pairs interacting over a short distance and, in particular, those closing hairpin loops, are predicted significantly better than long-range base pairs and those closing multistem loops and bulges. The prediction success of hairpin loops varies, however, with their size and context. Analysis of some of the RNAs that do not fold well suggests that the composition of some hairpin loops (e.g., tetraloops) and the higher frequency of noncanonical pairs in their comparatively derived structures might contribute to these lower success rates. Eucarya and mitochondrial rRNAs reveal further novel tetraloop motifs, URRG/A and CRRG, that interchange with known stable tetraloop in the procaryotes. PMID- 7489517 TI - Probing RNA tertiary structure: interhelical crosslinking of the hammerhead ribozyme. AB - Distinct structural models for the hammerhead ribozyme derived from single crystal X-ray diffraction and fluorescence resonance energy transfer (FRET) measurements have been compared. Both models predict the same overall geometry, a wishbone shape with helices II and III nearly colinear and helix I positioned close to helix II. However, the relative orientations of helices I and II are different. To establish whether one of the models represents a kinetically active structure, a new crosslinking procedure was developed in which helices I and II of hammerhead ribozymes were disulfide-crosslinked via the 2' positions of specific sugar residues. Crosslinking residues on helices I and II that are close according to the X-ray structure did not appreciably reduce the catalytic efficiency. In contrast, crosslinking residues closely situated according to the FRET model dramatically reduced the cleavage rate by at least three orders of magnitude. These correlations between catalytic efficiencies and spatial proximities are consistent with the X-ray structure. PMID- 7489518 TI - Prp16p, Slu7p, and Prp8p interact with the 3' splice site in two distinct stages during the second catalytic step of pre-mRNA splicing. AB - For the second catalytic step of pre-mRNA splicing to occur, a 3' splice site must be selected and juxtaposed with the 5' exon. Four proteins, Prp16p, Slu7p, Prp17p, Prp18p, and an integral spliceosomal protein, Prp8p, are known to be required for the second catalytic step. prp8-101, an allele of PRP8 defective in 3' splice site recognition, exhibits specific genetic interactions with mutant alleles of the other second step splicing factors. The prp8-101 mutation also results in decreased crosslinking of Prp8p to the 3' splice site. To determine the role of the step-two-specific proteins in 3' splice site recognition and in binding of Prp8p to the 3' splice site, we performed crosslinking studies in mutant and immunodepleted extracts. Our results suggest an ordered pathway in which, after the first catalytic step, Prp16p crosslinks strongly to the 3' splice site and Prp8p and Slu7p crosslink weakly. ATP hydrolysis by Prp16p affects a conformational change that reduces the crosslinking of Prp16p with the 3' splice site and allows stronger crosslinking of Prp8p and Slu7p. Thus, the 3' splice site appears to be recognized in two stages during the second step of splicing. Strong 3' splice site crosslinking of Prp8p and Slu7p also requires the functions of Prp17p and Prp18p. Therefore, Prp8p and Slu7p interact with the 3' splice site at the latest stage of splicing prior to the second catalytic step that can currently be defined, and may be at the active site. PMID- 7489519 TI - Identification of ribozymes within a ribozyme library that efficiently cleave a long substrate RNA. AB - Positions 2-6 of the substrate-binding internal guide sequence (IGS) of the L-21 Sca I form of the Tetrahymena thermophila intron were mutagenized to produce a GN5 IGS library. Ribozymes within the GN5 library capable of efficient cleavage of an 818-nt human immunodeficiency virus type 1 vif-vpr RNA, at 37 degrees C, were identified by ribozyme-catalyzed guanosine addition to the 3' cleavage product. Three ribozymes (IGS = GGGGCU, GGCUCC, and GUGGCU) within the GN5 library that actively cleaved the long substrate were characterized kinetically and compared to the wild-type ribozyme (GGAGGG) and two control ribozymes (GGAGUC and GGAGAU). The two control ribozymes have specific sites within the long substrate, but were not identified during screening of the library. Under single turnover conditions, ribozymes GGGGCU, GGCUCC, and GUGGCU cleaved the 818-nt substrate 4- to 200-fold faster than control ribozymes. Short cognate substrates, which should be structureless and therefore accessible to ribozyme binding, were cleaved at similar rates by all ribozymes except GGGGCU, which showed a fourfold rate enhancement. The rate of cleavage of long relative to short substrate under single-turnover conditions suggests that GGCUCC and GUGGCU were identified because of accessibility to their specific cleavage sites within the long substrate (substrate-specific effects), whereas GGGGCU was identified because of an enhanced rate of substrate binding despite a less accessible site in the long substrate. Even though screening was performed with 100-fold excess substrate (relative to total ribozyme), the rate of multiple-turnover catalysis did not contribute to identification of trans-cleaving ribozymes in the GN5 library. PMID- 7489520 TI - Purification and characterization of the Upf1 protein: a factor involved in translation and mRNA degradation. AB - mRNA degradation is an important control point in the regulation of gene expression and has been shown to be linked to the process of translation. One clear example of this linkage is the observation that nonsense mutations in a gene can accelerate the decay of the corresponding mRNA. In the yeast Saccharomyces cerevisiae, the product of the UPF1 gene, harboring zinc finger, NTP hydrolysis, and helicase motifs, was shown to be a trans-acting factor in this decay pathway. A UPF1 gene disruption results in stabilization of nonsense containing mRNAs and leads to a nonsense suppression phenotype. As a first step toward understanding the molecular and biochemical mechanism of nonsense-mediated mRNA decay, we have purified Upf1p from a yeast extract and characterized its nucleic acid-dependent NTPase activity, helicase activity, and nucleic acid binding properties. The results presented in this paper demonstrate that Upf1p contains both RNA- and DNA-dependent ATPase activities and RNA and DNA helicase activities. In the absence of ATP, Upf1p binds to single-stranded RNA or DNA, whereas hydrolysis of ATP facilitates its release from single-stranded nucleic acid. Based on these results, the role of Upf1p's biochemical activities in mRNA decay and translation are discussed. PMID- 7489521 TI - Alternative splicing of the Euglena gracilis chloroplast roaA transcript. AB - A novel gene, roaA (ribosomal operon-associated gene), encoding a potential RNA binding protein has been identified in the rpl23 ribosomal protein operon of the Euglena gracilis chloroplast genome. The roaA gene is interrupted by one group III and three group II introns. Introns 1 and 2 of roaA can be interpreted as a twintron, formed from the insertion of a group II intron into the 5'splice site of a group III intron. Alternative splicing of the group III intron results in two distinct transcripts encoding proteins of 516 and 514 amino acids. Group III introns may play a role in the generation of alternatively spliced transcripts. PMID- 7489522 TI - Minicircle-encoded guide RNAs from Crithidia fasciculata. AB - Although the mitochondrial uridine insertion/deletion, guide RNA (gRNA)-mediated type of RNA editing has been described in Crithidia fasciculata, no evidence for the encoding of gRNAs in the kinetoplast minicircle DNA has been presented. There has also been a question as to the capacity of the minicircle DNA in this species to encode the required variety of gRNAs, because the kinetoplast DNA from the C1 strain has been reported as essentially containing a single minicircle sequence class. To address this problem, the genomic and mature edited sequences of the MURF4 and RPS12 cryptogenes were determined and a gRNA library was constructed from mitochondrial RNA. Five specific gRNAs were identified, two of which edit blocks within the MURF4 mRNA, and three of which edit blocks within the RPS12 mRNA. The genes for these gRNAs are all localized with identical polarity within one of the two variable regions of specific minicircle molecules, approximately 60 bp from the "bend" region. These minicircles were found to represent minor sequence classes representing approximately 2% of the minicircle DNA population in the network. The major minicircle sequence class also encodes a gRNA at the same relative genomic location, but the editing role of this gRNA was not determined. These results confirm that kinetoplast minicircle DNA molecules in this species encode gRNAs, as is the case in other trypanosomatids, and suggest that the copy number of specific minicircle sequence classes can vary dramatically without an overall effect on the RNA editing system. PMID- 7489524 TI - Evaluation of cardiovascular grant-in-aid applications by peer review: influence of internal and external reviewers and committees. AB - OBJECTIVE: To determine what factors of the peer review process played a significant role in the assignment of scores for scientific merit by a major cardiovascular research finding agency. Specific variables studied included scores of internal and external reviewers, committee assignment and year of application. DESIGN AND SETTING: Retrospective analysis of research proposals submitted to the Heart and Stroke Foundation of Ontario from 1990 to 1994. SUBJECTS: Of a total database of 804 grant-in-aid (operating or project) proposals submitted to the Heart and Stroke Foundation of Ontario from 1990 to 1994, 779 had complete data and were included in the study. Variables analyzed were scores of internal and external reviewers, final committee score, committee assignment and year of application. MAIN RESULTS: There was poor correlation (r = 0.113) between external reviewers of grant proposals and only slightly better correlation between internal reviewers (r = 0.331). When mean internal and external scores were analyzed, correlation improved (r = 0.451) but agreement beyond chance was only moderate (weighed Cohen's kappa = 0.532). Regression analysis established that mean internal score had a greater impact on final committee score (r2 = 0.607, P < 0.001) than mean internal score (r2 = 0.348, P < 0.001). When review committee was entered as a dummy variable into a regression statement, it was statistically significant (P < 0.001) but explained less than 8% of variance in the final committee score. Mean scores showed a small 'upward creep' over the five-year period but explained less than 4% of variance in final committee score (r2 = 0.039, P < 0.001). CONCLUSIONS: In the peer review system studied, the scores of internal reviewers were more closely correlated to final committee score for scientific merit than those of external reviewers. Nevertheless, final committee scores were significantly different from either internal or external scores, suggesting that the process of committee discussion is an important, and highly influential, step in the peer review process. PMID- 7489523 TI - U12 snRNA in vertebrates: evolutionary conservation of 5' sequences implicated in splicing of pre-mRNAs containing a minor class of introns. AB - A minor class of introns with noncanonical splice sites has been identified in both vertebrate and invertebrate genomes. The divergent consensus sequences within these introns suggest that splicing might be via a mechanism distinct from that used by the major class of introns. The low abundance U12 snRNA has been proposed to base pair with the predicted branch site sequence of these minor class introns, probably bulging out an adenosine to act as the nucleophile in the first step of splicing. We have identified homologues of the previously characterized human U12 snRNA in both mouse and chicken, where the minor class of introns has also been found. The U12 sequences that potentially base pair with the putative branch site are invariant. Additional conserved sequences at the 5' end of U12 snRNA could dynamically base pair with U6 snRNA sequences flanking the hexanucleotide ACAGAG to form structures analogous to those of three U2-U6 interactions genetically defined as important in the major class of spliceosome. We have also isolated two human U12 snRNA genes. One gene is functional for transcription of U12 snRNA, whereas the other appears to be a pseudogene. Sequences of the 3' box in both U12 snRNA genes are strikingly similar and bear high resemblance to those of U1 and U2 genes. Upstream elements, including the PSE and the DSE, have been identified and characterized in the functional gene. These features indicate that transcription of U12 snRNA is driven by RNA polymerase II. PMID- 7489525 TI - Variations in cardiopulmonary resuscitation techniques: past, present and future. AB - OBJECTIVE: To review the major changes and advances in cardiopulmonary resuscitation (CPR) since its first description over 100 years ago. DATA SOURCES: A MEDLINE search of the English literature (1966 to 1994) was done using the key words 'cardiopulmonary resuscitation', 'CPR', 'heart massage', 'techniques' and 'methods'. STUDY SELECTION: Articles were selected based on content related to promising CPR techniques and specifically on alternate methods of closed chest compression. DATA EXTRACTION: Performed independently by the authors. DATA SYNTHESIS: Over the past three and a half decades, the importance of rapid institution of CPR has become increasingly recognized. This has led to a strenuous effort to educate the public in CPR delivery in addition to making definitive treatment with emergency medical systems more readily accessible. Furthermore, as the physiology of closed chest massage has become better understood, investigators have been able to focus their efforts appropriately to exploit the mechanisms that are operative during CPR. As a result, several modifications have been proposed in adjuvant therapies administered concomitantly with CPR, as well as in new, innovative methods of CPR delivery. Many of these changes and new methods show promise for improving vital organ perfusion and patient survival. CONCLUSIONS: CPR has become increasingly recognized as an important element in the chain of survival for victims of cardiopulmonary arrest. Several advances in delivery and technique have resulted in improved resuscitation of cardiopulmonary arrest victims. However, despite these advances, additional research is needed to improve further the still dismal overall survival of victims of cardiopulmonary arrest. PMID- 7489526 TI - Noninvasive testing in women presenting with chest pain: evidence for diagnostic uncertainty. AB - OBJECTIVE: To gain insight into the diagnostic utility of exercise stress testing in women. DESIGN: Observational prospective cohort study. SETTING: The Victoria General Hospital, Halifax, Nova Scotia. PARTICIPANTS: Consecutive women with chest pain referred by cardiologists or internists for exercise stress testing between May 30, 1992 and November 30, 1992 and followed prospectively to February 28, 1993. INTERVENTION: The proportion of patients subsequently referred for thallium scintigraphy and/or coronary angiography and their clinical profiles were determined. MAIN RESULTS: Of 183 patients studied, stress testing was positive in 48 (26.2%), negative in 48 (26.2%) and nondiagnostic in 87 (47.5%). Women with negative results were more likely to have had normal baseline electrocardiograms (ECGs) (P = 0.002) and least likely to have undergone prior angiography (P = 0.0003). Subsequent thallium scintigraphy and/or coronary angiography was undertaken in 33.3%, 18.8% and 27.6% with positive, negative and nondiagnostic index stress tests, respectively. None of chest pain, cardiac risk factors, previous cardiac investigations or baseline ECG discriminated 33 patients with negative or nondiagnostic stress results who had additional tests from 102 who did not. CONCLUSION: Exercise stress testing poorly screens women with chest pain for coronary artery disease (diagnostic in only 52.5%). Further study was undertaken in 27.6% with nondiagnostic tests and, surprisingly, in 18.8% with negative results. Why certain women with nondiagnostic, and so many with negative, stress tests were referred for further investigation was unclear. These results suggest diagnostic uncertainty when females presenting with chest pain are assessed. PMID- 7489527 TI - Comparison of the diagnostic yield of Holter versus transtelephonic monitoring. AB - OBJECTIVE: To compare the diagnostic yield of transtelephonic monitoring (TTM) with Holter monitoring in patients presenting possibly arrhythmogenic symptoms. DESIGN: A prospective comparison of Holter monitoring with TTM performed sequentially in all subjects. SETTING: Tertiary arrhythmia clinic at Hopital du Sacre-Coeur de Montreal, Montreal, Quebec. PATIENTS: One hundred consecutive patients (34 men and 66 women, mean +/- SD age 55 +/- 16 years) presenting over a two-year period for diagnosis of intermittent palpitations and/or syncope/dizziness. INTERVENTIONS: Subjects first underwent 24 h Holter monitoring and then were provided with TTM for 25 +/- 13 days, with instructions to record during symptomatic episodes and subsequently to transmit the recording at their convenience. MAIN RESULTS: Holter monitoring documented arrhythmia in 30 patients whereas TTM identified arrhythmia in 21. TTM was most useful in excluding arrhythmia during symptoms (34 patients) versus Holter (two patients). Neither method was useful in diagnosing syncope. Frequency of occurrence of palpitations did not predict which method would most likely yield a diagnosis but palpitations lasting longer than 2 mins were likely to be diagnosed by TTM. CONCLUSIONS: Holter and TTM are complementary studies whose combined use increases the diagnostic yield of arrhythmia. Further, TTM is of greatest use in excluding arrhythmia during intermittent symptoms. PMID- 7489528 TI - John Keith Lecture. Can cardiologists be cost effective? PMID- 7489530 TI - Comparative efficacy of two long-acting formulations of nifedipine in the treatment of hypertension. The Switch Study Investigators. AB - OBJECTIVE: To compare the antihypertensive effects and incidence of side effects of two formulations of nifedipine: a prolonged action (PA) tablet bid and a gastrointestinal therapeutic system (GITS) tablet once daily. DESIGN: Patients with controlled hypertension (diastolic blood pressure less than 90 mmHg) on nifedipine-PA 10 mg (n = 74) or 20 mg (n = 99) bid were enrolled into an open label study consisting of six weeks of previous nifedipine-PA followed by six weeks of nifedipine-GITS therapy. Nifedipine-GITS was increased from 30 mg to 60 mg daily if the patient's diastolic blood pressure increased by at least 10 mmHg compared with the value after six weeks of nifedipine-PA. MAIN RESULTS: Blood pressure after six weeks of nifedipine-PA 10 mg (142 +/- 1/86 +/- 1 mmHg) decreased (P < 0.01) after conversion to nifedipine-GITS 30 mg (137 +/- 1/84 +/- 1 mmHg). Blood pressure values for nifedipine-PA 20 mg patients were unchanged after conversion to nifedipine-GITS; 74 patients received nifedipine-GITS 30 mg and 14 patients required titration up to nifedipine-GITS 60 mg daily. Sixteen patients withdrew on nifedipine-PA and five withdrew on nifedipine-GITS, mostly for reasons unrelated to drug therapy. Twenty-three patients experienced adverse events on nifedipine-PA versus 20 patients on nifedipine-GITS therapy. CONCLUSIONS: Patients whose hypertension is controlled by nifedipine-PA 10 or 20 mg bid can be successfully converted to nifedipine-GITS with most patients remaining normotensive on 30 mg daily. PMID- 7489529 TI - Walking program reduces elevated cholesterol in women postmenopause. AB - OBJECTIVE: To examine the effect of a moderate exercise regimen on total serum cholesterol, triglyceride, high density lipoprotein cholesterol (HDL-C), low density lipoprotein cholesterol (LDL-C), body composition and cardiovascular fitness (VO2 max) in mildly hyperlipidemic women, postmenopause. DESIGN: Randomized assignment to walking (n = 24) or control (n = 16) groups. SETTING: Community based intervention. PARTICIPANTS: Over 300 volunteers were screened to obtain the sample of 40 eligible women. Participants were mildly hyperlipidemic, postmenopause (mean age 62.0 +/- 5.7 years), sedentary, nonsmokers and not on hormone replacement therapy. Results are reported for the 25 subjects (15 walkers, 10 controls) who completed the study. INTERVENTION: Exercisers walked an average of 54.3 +/- 7.7 mins/day, 4.9 +/- 1.7 days/week, at an intensity of 54% maximum heart rate reserve, for six months. Participants were counselled not to change their diets. MAIN RESULTS: Total serum cholesterol, triglyceride, total serum cholesterol: HDL-C ratio, weight and fat mass decreased significantly in the walkers compared with the controls (P < 0.05), as did body mass index (P < 0.01). Walking resulted in a significant increase in VO2 max (P < 0.01). Changes in serum lipids were significantly related to changes in body fat, but not to change in aerobic fitness. There were no changes in dietary intake. CONCLUSIONS: A moderate intensity exercise program induces favourable alterations in total serum cholesterol and other atherogenic indices in hyperlipidemic women postmenopause, and these changes are related more to loss of body fat than to increased fitness level. PMID- 7489531 TI - Successful treatment of supraventricular tachycardia-induced cardiomyopathy with amiodarone: case report and review of literature. AB - Permanent junctional reciprocating tachycardia (PJRT) is a rare cause of supraventricular tachycardia in the pediatric population and is resistant to most pharmacological therapy. A case of supraventricular tachycardia that, on the basis of postnatal electrocardiographic and Holter monitor evidence, was diagnosed as PJRT and presented in utero as an atypical tachycardia with severe tachycardia-induced cardiomyopathy confirmed postnatally is presented. It is the only case of which the authors are aware that was controlled in the neonatal period by amiodarone and that resulted in complete resolution of systolic dysfunction. The literature discussing how tachycardia may induce cardiomyopathy and the use of amiodarone in treatment both pre- and postnatally are reviewed. PMID- 7489532 TI - Spontaneous pericardial tamponade during PTCA. AB - A case of acute hemorrhagic pericardial tamponade complicating a successful percutaneous transluminal coronary angioplasty (PTCA) is described, in the setting of rheumatoid arthritis (with no evidence of prior or concomitant pericarditis), large doses of intravenous heparin administration and a relatively high activated clotting time. There was no evidence of coronary artery rupture and there was no recent use of other anticoagulants or thrombolytic agents. Successful treatment comprised emergency pericardial drainage and intravenous protamine sulphate. The authors believe this to be the first reported case of acute hemorrhagic pericardial tamponade due solely to heparin administration. The possibility of acute hemorrhage of a rheumatoid nodule was considered but subsequent magnetic resonance imaging scan with contrast gadolinium was normal. PMID- 7489533 TI - Ventricular arrhythmia complicating Kawasaki disease. AB - Cardiac sequelae account for the major morbidity and mortality related to Kawasaki disease, and subclinical myocarditis is a frequent finding. The unusual finding of ventricular arrhythmia in a 2.5-year-old patient during the acute phase of Kawasaki disease is reported. The arrhythmia resolved during empirical treatment with oral steroids. PMID- 7489534 TI - Effect of perindopril in pacing-induced canine models of acute and chronic heart failure. AB - OBJECTIVE: To examine the effects of perindopril, a nonsulfhydryl-containing angiotensin-converting enzyme inhibitor, on total vascular capacitance and hemodynamics in acute and chronic dog models of heart failure. METHODS: Acute heart failure was induced in anesthetized, splenectomized dogs by a volume load (dextran 70, 20 mL/kg) during rapid right ventricular pacing (RRVP) at 250 beats/min. Pretreatment with perindopril (0.3 mg/kg daily for six days, n = 7) was compared with no treatment (n = 7). Total vascular capacitance and compliance were measured from plots of mean circulatory filling pressure during acetylcholine-induced circulatory arrests at different blood volumes. Chronic heart failure was induced by continuous RRVP in splenectomized dogs treated with perindopril (0.3 mg/kg daily, n = 8), which were compared with untreated dogs (n = 8). Hemodynamics and total vascular capacitance and compliance were measured at baseline and after 33 days of RRVP. RESULTS: Perindopril treatment did not significantly modify the increased pulmonary capillary wedge and mean circulatory filling pressures, reduced total vascular compliance or total vascular capacitance associated with the volume load and acute RRVP. During chronic RRVP, perindopril reduced weight gain and the development of ascites, reduced right atrial pressure (6.3 +/- 1.3 versus 10.3 +/- 1.2 mmHg), mean circulatory filling pressure (9.3 +/- 1.0 versus 14.7 +/- 1.2 mmHg), stressed blood volume (22 +/- 3 versus 33 +/- 4 mL/kg) and central blood volume (10 +/- 1 versus 14 +/- 1 mL/kg) while increasing cardiac output (122 +/- 9 versus 98 +/- 7 mL/kg). However, the reduction in total vascular capacitance was not attenuated and pulmonary capillary wedge pressure was not lowered significantly (18.5 +/- 1.5 versus 21.4 +/- 1.3 mmHg). CONCLUSION: Perindopril failed to modify hemodynamics in the pacing-induced canine model of acute heart failure but had beneficial effects in the model of chronic heart failure. PMID- 7489536 TI - Effect of calcium antagonists in variant or Prinzmetal angina. AB - Variant angina, first described by Prinzmetal in 1959, is a syndrome of recurrent ischemic-type chest pain that occurs at rest and is associated with ST segment elevation. The syndrome has been convincingly demonstrated to be caused by temporary occlusion at a site of local spasm. The objectives of this article are to review treatment of the patient with variant angina and to address the important role of calcium antagonists. PMID- 7489535 TI - Sex-related alterations in antioxidant status and susceptibility to atherosclerosis in Japanese quail. AB - OBJECTIVE: To examine antioxidant enzyme activities in aorta and blood of male and female Japanese quail on control or cholesterol-supplemented diets, given the strong influence of sex and the postulated role of oxidative processes in the pathogenesis of atherosclerosis. ANIMALS AND METHODS: Atherosclerosis-prone Japanese quail (15 of each sex) were either maintained on a control diet or fed a cholesterol-supplemented (1% by weight) diet for nine weeks. RESULTS: Plasma cholesterol levels in controls were comparable in both sexes, although females showed higher triglyceride levels. Cholesterol supplementation increased plasma cholesterol in both males (8.1-fold) and females (2.5-fold) but triglycerides were significantly elevated (3.4-fold) by cholesterol feeding only in males. Cholesterol supplementation increased aortic cholesterol content to a greater extent in males (2.2-fold) than in females (1.4-fold) and plaque formation was apparent only in males. Cholesterol-fed birds showed small, but significant, sex related differences in antioxidant enzyme profiles of aorta (glutathione peroxidase activity was higher in males and superoxide dismutase activity was lower in females) and red blood cells (glutathione peroxidase activity was higher in females). Cholesterol feeding was associated with opposing effects on the activity of plasma glutathione peroxidase (increased in males but decreased in females). Evidence was also found for complex sex- and tissue-related differences in patterns of ageing over the nine-week experimental period. CONCLUSIONS: Although sex-related differences in antioxidant enzyme profiles are demonstrable in aortic tissues and blood of Japanese quail fed control or cholesterol supplemented diets, it seems unlikely that these are of sufficient magnitude to account for the large difference in susceptibility to atherosclerosis between males and females. The complex sex- and tissue-dependent effects of ageing on the activities of antioxidant enzymes observed emphasize the importance of age- and sex-matched controls in experimental studies of antioxidant enzyme alterations in disease processes generally. PMID- 7489537 TI - Blalock and Taussig--1949. PMID- 7489538 TI - Rules of evidence and clinical recommendations. PMID- 7489539 TI - Self-measurement of blood pressure: benefits, risks and interpretation of readings. The Canadian Coalition for High Blood Pressure Prevention and Control. AB - The literature on the benefits and risks of self-measurement of blood pressure and interpretation of the readings was reviewed. Self-measurement of blood pressure is useful in selected patients with high blood pressure, can be used to monitor blood pressure closely outside health care facilities, and can determine whether patients have white coat hypertension. The involvement of patients in their own blood pressure management is increased and self measurement may result in more rapid achievement of target blood pressure readings, improvement in adherence to antihypertensive therapy and decreased health care utilization. However, some patients may not be suited to monitor their own blood pressure and some may experience an increase in anxiety regarding their health. With careful training and selection of patients, most can accurately assess their blood pressure. Self-measured readings are generally lower than readings in a physician's clinic (or office) and this must be accounted for in assessing response to therapy and usual levels of blood pressure. Self-measured readings are a valuable supplement to clinic readings in many patients. PMID- 7489540 TI - Self-measurement of blood pressure: issues related to the training of patients. Canadian Coalition for High Blood Pressure Prevention and Control. AB - For effective self-measurement of blood pressure, patients need to be able to obtain accurate and reproducible measurements. Criteria for measuring blood pressure have been well developed in research studies, and through the use of standardized training programs it has been established that nonmedically trained individuals can obtain valid measurements. These recommendations for blood pressure measurement were adapted to the self-measurement situation. Either manual sphygmomanometers or automated electronic devices can be used effectively. Although automated devices may be easier to use, the health care practitioner should ensure that any such device has been demonstrated to be accurate and reliable before its adoption. There are no reported programs available for training individuals in the use of self-measurement. A check-list has been developed for use by health care practitioners. It is estimated that proper training will usually take more than 20 mins. Since many health care practitioners do not use the recommended blood pressure measurement technique, there is also a need to develop programs to 'train the trainers'. Finally, individuals undertaking regular self-measurement should have their technique and the calibration of their instruments checked at regular intervals (six to 12 months). PMID- 7489541 TI - Self-measurement of blood pressure: assessment of equipment. Canadian Coalition for High Blood Pressure Prevention and Control. AB - Equipment for the self-measurement of blood pressure is readily available to consumers. These devices use one or more surrogate (indirect) measures of pressure to estimate systolic and diastolic blood pressure. Manual auscultatory devices using stethoscope and sphygmomanometer have been adapted for home use, but a variety of automated devices based on auscultation, oscillometry, and other techniques are available and may be more suitable for individuals who have limited vision, hearing or dexterity. Despite the existence of voluntary evaluation protocols and mandatory manufacturing standards, blood pressure readings from some automatic devices may not be accurate. Some devices are packaged with insufficient information to ensure proper use, and most individuals need some form of guidance in their use and calibration testing. If self measurement of blood pressure is to be of benefit, the health care professional must recommend only those devices that are accurate and suitable to the patient or client. The Canadian Coalition for High Blood Pressure Prevention and Control will endeavour to develop a regular means by which health care professionals can keep informed of available devices for blood pressure self-measurement. PMID- 7489542 TI - Blood pressure measurement. AB - Cardiovascular diseases account for over half of all deaths in industrialized societies, and hypertension is one of the most treatable and preventable risk factors underlying that impressive statistic. Implicit in population and community control programs for high blood pressure is the recognition, evaluation and treatment of patients with elevated arterial pressures. The indirect measurement of arterial pressure has been available only within the past century, and the definition of procedures and techniques for its widespread use have been introduced only within the past few decades. The development of guidelines and recommendations for sphygmomanometry has been aided by epidemiological criteria and national hypertension programs worldwide. These guidelines have permitted identification of large numbers of patients with hypertension. Implicit in the screening of populations is the necessity to confirm the elevated arterial pressure on at least three separate occasions before diagnosis and treatment are instituted (unless the clinician deems that a hypertensive urgency or emergency exists). A number of pitfalls and practical considerations are discussed, and pertinent references to currently published guidelines are presented. PMID- 7489544 TI - Utility of 24 h ambulatory blood pressure monitoring in clinical practice. AB - The technique of noninvasive ambulatory blood pressure monitoring was first described more than 30 years ago, but is only now becoming accepted as a clinically useful procedure for the evaluation of patients with abnormal blood pressure regulation. This applies mostly to those with known or suspected high blood pressure, but also includes the less common situation of low blood pressure. In addition to providing a better estimate of the average or true blood pressure, ambulatory monitoring may reveal changes in the variations in blood pressure over 24 h that would not be detected by more conventional measurement techniques. The two most important aspects are alterations in the pattern of the diurnal rhythm of blood pressure and short term blood pressure variability. PMID- 7489543 TI - A curriculum for the training and certification of blood pressure measurement for health care providers. AB - Blood pressure measurement in clinical practice is almost never performed according to accepted guidelines. This is likely to be a result of inadequate training. We describe a standardized training program for teaching blood pressure measurement to health care students, practitioners and clinical investigators as well as lay personnel. The goal is to have the student understand the rationale and be able to perform accurate blood pressure measurements. Students must master the knowledge of proper blood pressure measurement technique, document their accuracy on standardized video tests and demonstrate their performance of proper technique. In addition the program reviews key historical and physiological aspects of blood pressure measurement and the cardiovascular complications of high blood pressure. The operation and maintenance of sphygmomanometers and the stethoscope, and monitoring for biases in blood pressure measurements in clinical practice, are also covered. The curriculum time requires a minimum of 7 h and biannual recertification is recommended. With training of instructors, the program can be easily replicated and should be considered in any setting in which blood pressure measurement is taught. PMID- 7489545 TI - Self-measurement of blood pressure: recommendations of the Canadian Coalition for High Blood Pressure Prevention and Control. AB - OBJECTIVE: To provide health care professionals with guidelines on the use of blood pressure self-measurement. METHODS: Recommendations were devised after consideration of expert reviews and guidelines, personal files, international standards documents, personal communication with investigators and the results of a MEDLINE search (1966-94) using the term 'blood pressure determination'. BENEFITS, HARMS, COSTS: Self-measurement of blood pressure can be used to detect white coat hypertension, monitor changes in blood pressure closely, more rapidly achieve desired blood pressure goals, increase adherence to antihypertensive therapy and improve patient self-reliance. However, self-measured blood pressure readings may be misleading because there is insufficient normative, prognostic and outcome data and because some patients may not take accurate measurements. The use of self-measurement of blood pressure has a relatively small direct cost and may result in an overall reduction in treatment costs. RECOMMENDATIONS: Self measured blood pressure readings can be a valuable supplement to clinic (or office) blood pressure readings. However, self-measurement is appropriate neither for patients who are physically or mentally incapable of accurate assessment and interpretation of readings nor for those who do not want to participate. Patients who self-monitor blood pressure require careful training in blood pressure measurement and instruction on the recording and interpretation of blood pressure readings. Advice to patients using monitoring equipment must take into account the needs and abilities of the patient. Although only a few electronic devices for the self-measurement of blood pressure have met recommended performance standards, their use may be more appropriate for some patients and the training requirements fewer than if manual devices are used. VALIDATION: The guidelines of several expert groups were examined in the preparation of these recommendations. The recommendations were presented at the World Conference on Hypertension Control in 1995 and were reviewed by the parent societies of the Canadian Coalition for High Blood Pressure Prevention and Control. PMID- 7489546 TI - Anesthetists taking drugs: is overwork the cause? PMID- 7489547 TI - Effect of unemployment on health: acknowledgement. PMID- 7489548 TI - Did continuous quality improvement lead to lower episiotomy rate? PMID- 7489549 TI - Refusal to police breast-feeding. PMID- 7489551 TI - Could changes in the wheelchair delivery system improve safety? AB - Despite emerging evidence about the high incidence and severity of wheelchair related injuries, regulations governing wheelchair safety are almost nonexistent in Canada. The authors believe that, to improve wheelchair safety, a concerted effort by government, manufacturers, purchasing groups, users and clinicians is needed. Health Canada's Health Protection Branch should treat wheelchairs as medical devices (as defined in the Food and Drugs Act 1985) and improve its injury-reporting network. Manufacturers should give a higher priority to safety in wheelchair design, improve their educational materials and formalize postmarketing surveillance. Purchasing groups should try to ensure that they do not stifle innovation in wheelchair design by setting unrealistic reimbursement ceilings and should use their market power more effectively. Users should obtain their wheelchairs in specialized settings, heed safety warnings and make more effective use of litigation when such action is warranted. Clinicians should ensure that patients are equipped with the most appropriate wheelchair for their needs, that they are given adequate training in safe wheelchair use and that they understand the dangers involved. Rapid changes in wheelchair technology and emerging evidence about the high incidence and severity of injuries related to wheelchair use suggest that such changes are needed in the wheelchair delivery system. PMID- 7489550 TI - Evaluation of clinical practice guidelines. AB - Compared with the current focus on the development of clinical practice guidelines the effort devoted to their evaluation is meagre. Yet the ultimate success of guidelines depends on routine evaluation. Three types of evaluation are identified: evaluation of guidelines under development and before dissemination and implementation, evaluation of health care programs in which guidelines play a central role, and scientific evaluation, through studies that provide the scientific knowledge base for further evolution of guidelines. Identification of evaluation and program goals, evaluation design and a framework for evaluation planning are discussed. PMID- 7489552 TI - Requirements for admission to medical school: how many years of university study are necessary? AB - OBJECTIVE: To assess whether students admitted to medical school after completing 2 years of undergraduate study performed as well as those admitted after longer periods of undergraduate study in terms of broad patient-care skills measured at the time of graduation. DESIGN: Retrospective study. SETTING: University of Alberta, Edmonton. PARTICIPANTS: Graduates of the classes of 1990 and 1991, of the 226 graduates 133 had entered medical school after 2 years of undergraduate training, 39 after 3 years and 54 after 4 or more years. Eight students had been excluded because they were either transfer students or international students. OUTCOME MEASURES: Objective and subjective assessments of the main clinical rotations (internal medicine, obstetrics and gynecology, pediatrics, psychiatry, radiology and surgery), results of the faculty's final comprehensive examination and of the Medical Council of Canada's Qualifying Examination. RESULTS: The students who had completed 2 years of undergraduate study before medical school were significantly younger than those who had completed 3 years and those who had completed 4 or more years (mean age [and standard deviation (SD)] 20.5 [2.1], 21.5 [2.4] and 25.1 [4.4] years respectively, p < 0.001). They also had a significantly higher mean grade point average (GPA) for the prerequisite courses for admission to medical school than those with 3 years and those with 4 or more years of undergraduate study (8.26 [SD 0.3], 7.95 [SD 0.3] and 7.80 [SD 0.5] respectively, p < 0.001). The overall mean GPA for the best 2 years of undergraduate study did not differ significantly between the three groups. The students with 2 years of undergraduate study had a significantly lower mean score for the pre-entry interview than those who had 4 or more years of undergraduate study (32.1 [SD 7.6] v. 38.3 [SD 8.5], p < 0.001). There were no significant differences between the three groups in the results of any of the subjective or objective outcome measures. CONCLUSION: Students who completed 2 years of undergraduate study before admission to medical school were able to achieve a satisfactory level of competency and maturity by the end of medical school. The 2 year option for entrance into medical school should be reconsidered. PMID- 7489553 TI - Voluntary anonymous linked study of the prevalence of HIV infection and hepatitis C among inmates in a Canadian federal penitentiary for women. AB - OBJECTIVE: To determine the seroprevalence of HIV infection and hepatitis C among inmates of a federal penitentiary for women. DESIGN: Voluntary, anonymous, linked, point-prevalence study involving testing of blood samples for antibodies to HIV and hepatitis C virus. PARTICIPANTS: All inmates of the multilevel security federal Prison for Women, Kingston, Ont., who volunteered to participate in the study. Inmates at this long-stay facility are from across Canada. OUTCOME MEASURE: Seroprevalence rate among participants of antibodies to HIV and hepatitis C virus. RESULTS: Of the 130 inmates available for study 113 (86.9%) agreed to donate a blood sample. One woman (0.9%) was HIV positive; 45 (39.8%) were positive for hepatitis C antibody. CONCLUSIONS: It is possible to obtain a high participation rate in a voluntary, anonymous, linked point-prevalence study in a long-stay penitentiary. The HIV seroprevalence rate of 0.9% is lower than that found in studies in provincial (short-stay) prisons. However, the high rate of antibodies to hepatitis C suggests a significant level of risk behaviour, most likely injection drug use, and suggests the potential for a rapid increase in the rate of HIV infection should the number of newly admitted HIV-positive inmates who use injection drugs rise. PMID- 7489554 TI - HIV: evolution of a pandemic. AB - Although the prevalence of AIDS is still relatively low in many countries in Asia and the Pacific Rim, the rate of HIV transmission in this region continues to rise inexorably and will surpass that of sub-Saharan Africa by 1997. The challenge of mobilizing governments and communities to counter this largely invisible threat was the theme of the Third International Conference on AIDS in Asia and the Pacific, held in Chiang Mai, Thailand, in September 1995. Thailand has led the way with bold and far-reaching HIV prevention programs. Nevertheless, the long-term consequences of existing HIV infection in Thailand and elsewhere in Asia will be severe. Moreover, these repercussions will be felt globally as productivity is undermined, health care costs soar and purchasing power weakens. Supporting programs for HIV prevention and care abroad is thus an urgent matter of economic and political self-interest as well as a humanitarian imperative. PMID- 7489555 TI - Texas beckons. AB - Many Canadian medical families have been approached by US headhunters anxious to recruit family physicians to practise in the US. Lynne Sears Williams and her husband, Calgary family physician Jim Williams, went to Texas to see what opportunities awaited. In this article she describes the high and low points of recruitment efforts south of the border. PMID- 7489556 TI - Computers a cornerstone of evidence-based care, conference told. AB - Computers are poised to become key players in the delivery of health care, but are physicians ready for them? A recent conference on medical communication in the electronic era examined the potential of computers to assist in diagnosis, provide continuing medical education, disseminate evidence and research findings, and simplify practice management. However, delegates were told that even though many medical practices are computerized, it is often staff members and not physicians who use the technology. For computers to gain wider acceptance for medical purposes, physicians need to be made comfortable with their use at an early stage of training. PMID- 7489557 TI - Consensus finally achieved on resuscitative interventions. AB - A joint policy statement on the resuscitation of patients is published in this issue of CMAJ. Dr. John Williams, the CMA's director of ethics and legal affairs, discusses how it differs from the joint statement published last year. PMID- 7489558 TI - Canadians pay for harsh economic policies with their health, physicians warn. AB - Canada's fiscal policies are damaging the health of Canadians, two physicians told a conference that examined globalization's impact on the country. Near record unemployment levels and the recent recession have forced 41% of families in which the parents are 30 or younger to live below the poverty level; more than 21% of Canadian children are also considered to live in poverty. The impact tight fiscal policies have on health and well-being are enormous, say the dean of medicine at the University of Western Ontario and the chair of the Canadian Institute of Child Health. PMID- 7489559 TI - US report on violence in the medical workplace may hold lessons for Canadian MDs. AB - Concern about the growing incidence of violence against health care professionals has prompted the Young Physicians Section of the American Medical Association to research and develop a publication designed to raise physicians' awareness. Violence in the Medical Workplace. Prevention Strategies provides some commonsense precautions that could minimize the possibility a physician will fall victim to violence at the hands of a patient. PMID- 7489560 TI - Joint statement on resuscitative interventions (update 1995). CMA policy summary. AB - This joint statement includes: guiding principles for health care facilities when developing cardiopulmonary-resuscitation (CPR) policy; CPR as a treatment option; competence; the treatment decision, its communication, implementation and review; and palliative care and other treatment. This joint statement was approved by the Canadian Healthcare Association, the CMA, the Canadian Nurses Association and the Catholic Health Association of Canada and was developed in cooperation with the Canadian Bar Association. PMID- 7489561 TI - At one time, operations outside the hospital were not uncommon. AB - Before World War II, it was not uncommon for a person with influenza caused by streptococcus to develop empyema. Retired surgeon Harry Morton recalls two well known patients who had empyema-related operations outside the hospital. PMID- 7489562 TI - Splenectomy before tumor inoculation prolongs the survival time of cachectic mice. AB - The effects of splenectomy on the development of cachexia, tumor growth and animal survival were studied in tumor-bearing CDF1 mice. Mice were inoculated with two subclones of colon 26 adenocarcinoma, clone 20 (with a potent capacity to induce cachexia) and clone 5 (without such activity), and underwent splenectomy before or after tumor inoculation. Splenectomy significantly prolonged the survival of mice bearing clone 20 when it was performed prior to tumor inoculation, although the progression of cachexia and tumor growth were not affected. The survival rate was higher in splenectomized than it was in nonsplenectomized mice 20-40 days after tumor inoculation. Such effects on survival were not observed, however, in mice splenectomized after inoculation with clone 20 or in mice that underwent splenectomy either before or after inoculation with clone 5. The decrease of peripheral blood lymphocyte count observed in mice bearing clone 20 was magnified when splenectomy was performed before tumor inoculation, but the serum levels of tumor necrosis factor and interleukin-6 were comparable. These results indicate that cancer death from cachexia is not directly attributable to enhanced catabolism. The mechanism by which splenectomy ameliorates the survival of cachectic mice remains to be studied, although several changes observed in the splenectomized mice after inoculation, including decreases in the peripheral blood L3T4+ cells and Lyt-2+ cells on the 9th day and 15th day respectively, and increase in the L3T4+/Lyt-2+ cell ratio on the 15th day suggest the involvement of the modified host's immune response. PMID- 7489563 TI - CD4+ Th0 cell clones, isolated from a metastatic lymph node of a melanoma patient, possess cytolytic function. AB - In the present study T lymphocytes isolated from a metastatic lymph node (T-LNL) of a melanoma patient have been cloned. In the attempt to verify whether T-LNL may acquire in vitro functional activities in the absence of tumour-associated antigens, they were cloned utilizing allogenic lymphocytes as feeder cells. Nineteen clones generated from T-LNL proved to be CD4+ and, among these, five were able to kill autologous and allogeneic human melanoma cells in HLA-class-II restricted way. On the basis of their cytokine production, these CD4+ cytolytic T LNL clones were shown to belong to the Th0 subset and three of them expressed the V beta 17 chain of the T cell receptor. These results suggest the presence of melanoma-specific but functionally inactive lymphocytes with T cell receptor oligoclonality in the lymph node environment. These specific T cells may acquire in vitro the capacity to kill autologous and allogeneic tumours without any induction by autologous melanoma cells. PMID- 7489565 TI - Monocyte chemoattractant protein-1 (MCP-1) gene transduction: an effective tumor vaccine strategy for non-intracranial tumors. AB - Recently, there has been renewed interest in the concept of tumor vaccines using genetically engineered tumor cells expressing a variety of cytokines to increase their immunogenicity. Human MCP-1 (JE) is a potent chemoattractant and activator of monocytes and T lymphocytes and thus a good candidate gene for a tumor vaccine. We therefore evaluated the efficacy of vaccines consisting of irradiated tumor cells transduced with the murine MCP-1 gene in the syngeneic 9L gliosarcoma brain tumor model. 9L cell lines stably expressing murine MCP-1 (9L-JE) and control cell lines expressing neomycin 3' phosphotransferase (9L-Neo) were generated by infection with a Moloney murine leukemia retroviral vector. Fisher 344 rats were immunized with intradermal injections of 5 x 10(5) or 2 x 10(6) irradiated (5000 cGy) 9L-JE, 9L-Neo, and wild-type 9L (9L-WT) cells. Two weeks later immunized and non-immunized animals were challenged with various doses of intradermal (5 x 10(6)-5 x 10(7) or intracerebral (2 x 10(4)-5 x 10(5) 9L-WT cells. Intradermal tumors grew in all non-immunized animals. No tumors grew in animals immunized with irradiated 9L-JE or 9L-Neo cells and challenged with inocula of fewer than 5 x 10(5) 9L-WT cells. With higher inocula up to 10(7) cells, tumors appeared in all the animals, but subsequently regressed in the immunized animals. Tumors in animals immunized with 9L-JE were always smaller than tumors in the other groups. In addition, only the 9L-JE vaccine protected against tumor inocula of 5 x 10(7) cells. Thus vaccination with MCP-1-expressing cells was able to protect animals against at least a 100-fold larger number of challenge tumor cells than vaccination with control cells. In contrast to studies with intradermal tumors, immunization with 9L-JE and 9L-Neo produced only minimal protection against intracerebral tumors. There was no significant difference between the 9L-JE and 9L-Neo vaccines in intracerebral challenge. This study suggests that tumor vaccines expressing cytokine genes such as MCP-1 can increase the antitumor response. However, the protective effect of these vaccines appears to be largely limited to intradermal tumors rather than intracerebral tumors. PMID- 7489564 TI - Immunogenicity and immunosensitivity of ex vivo human carcinomas: interferon gamma and tumour necrosis factor alpha treatment of tumour cells potentiates their interaction with autologous blood lymphocytes. AB - Human carcinoma cells vary appreciably in the expression of MHC class I, class II, ICAM-1 (CD54) and B7 (CD80) molecules. Short-term in vitro exposure of ex vivo carcinoma cells to interferon gamma and tumour necrosis factor alpha elevated/induced the surface expression of MHC class I, class II and ICAM-1, but only rarely of B7. We found that cytokine treatment elevated the cytotoxic susceptibility and the stimulatory potential of ex vivo tumour cells. This was demonstrated (a) by the increased frequency and elevated level of auto-tumour lysis and (b) by induction of DNA synthesis and generation of cytotoxic lymphocytes in autologous mixed lymphocyte/tumour cell culture (MLTC). The MHC class I and ICAM-1 molecules on the tumour cells were required for interaction with the lymphocytes as indicated by the inhibitory effect of specific mAb both in the stimulation and in the cytotoxic tests. While the cytokine-induced increases in MHC and ICAM-1 on the low-expression tumours were probably important for the modification of functional interaction with the autologous lymphocytes, it is likely that alterations in other properties of tumour cells were also induced which contributed to the phenomenon. This was indicated by the results obtained with several tumours, which expressed indigenously high levels of these molecules but activated the autologous lymphocytes only after cytokine treatment. In several experiments the untreated targets that did not activate the lymphocytes were sensitive to the cytotoxicity of the effectors activated in MLTC. The results show that the afferent and efferent arms of the immune response have different requirements for functional interactions between lymphocytes and tumour cells. PMID- 7489566 TI - Induction of tumor immunity by intact irradiated leukemic B cells (BCL1) bearing a tumor-associated cell-surface idiotype and the costimulatory B7 molecule. AB - The idioptypic (Id) determinant of immunoglobulin expressed on the cell surface of malignant B cells represents a prototypical tumor-associated antigen (TAA), which has been used in a purified soluble form for active immunization in experimental tumor models and human hematological malignancies. Using a spontaneous transplantable murine model of B cell leukemia/lymphoma (BCL1), we have demonstrated the expression of the B7 costimulatory molecules in addition to the previously described Id determinant and class II major histocompatibility antigens. Intact irradiated BCL1 cells bearing these distinct determinants induced long lasting antitumor immunity in naive syngeneic mice. Induction was dose-dependent and most effective when three doses of 30 x 10(6) intact irradiated BCL1 cells were given at intervals of 7-10 days. The induced immunity protected 96% of 28 mice inoculated with a lethal dose of 10(5)-10(6) nonirradiated BCL1 cells and 85% of 27 mice given a second challenge, whereas control mice died on day 20 after inoculation with 10(6) BCL1 cells. Adoptive transfer of splenocytes derived from immune mice did not induce leukemia in syngeneic recipients. Such splenocytes, harvested more than 365 days following immunization and administered together with fresh BCL1 cells to adoptive recipients, were able to confer protection for 90 days, even following a second challenge given 104 days after the first one. BCL1 immune splenocytes transferred into BCL1-bearing mice exerted a therapeutic effect, preventing leukemia onset for at least 180 days. Our results demonstrate the ability of tumor cells to trigger effective anti-tumor immunity. These findings could ultimately be applied to the prevention of tumor relapse in treatment of hematological and other malignancies expressing TAA, class II MHC antigen and costimulatory molecules. PMID- 7489567 TI - Tumour-infiltrating lymphocytes mediate lysis of autologous squamous cell carcinomas of the head and neck. AB - Tumour-infiltrating lymphocytes (TIL) and tumours from six patients with squamous cell carcinomas of the head and neck (SCCHN) were investigated. The six tumours all expressed major histocompatibility complex (MHC) class I antigens both in vivo and as tumor cell lines grown in vitro. In addition, the cancer cells either overexpressed the tumour-suppressor gene product p53 or harboured human papilloma virus 16/18 (HPV). The TIL were expanded in vitro in the presence of interleukin 2, immobilised anti-CD3 mAb and soluble anti-CD28 mAb. Expanded TIL cultures contained both CD4+ and CD8+ T cells, but generally contained few CD56+CD3- cells of the natural killer (NK) phenotype. CD8+ T cells dominated the individual TIL cultures from five of the six patients and showed significant autologous tumour cell lysis. In TIL cultures derived from four of these tumour-reactive TIL cultures, killing could be partially blocked by an anti-MHC class I mAb. TIL cultures reacting with autologous tumour cells also showed strong TCR/CD3 redirected cytotoxicity when assayed against hybridoma cells expressing anti TCR/CD3 mAb as well as natural-killer(NK)-like activity. A number of TIL cultures devoid of autologous tumour cell lysis were capable of lysing the natural killer(NK)-sensitive K562 cell line suggesting that the SCCHN cells themselves are resistant to NK-like lysis. In conclusion, TIL cultures from head and neck carcinomas contain T cells which, upon expansion in vitro, can lyse autologous tumour cells in a MHC-class-I-restricted fashion. Thus, the results of the present study document that carcinomas of the head and neck in some patients are infiltrated by cytotoxic T cell precursors potentially capable of rejecting the autologous tumour. PMID- 7489568 TI - An indicator quantitatively comparing two treatment effect sizes on responder and non-responder groups--exponential of estimated interaction parameter. AB - Sometimes a specific treatment is effective in one subgroup but not in another. An indicator allowing quantitative comparison of treatment effect in two subgroups would be useful in clinical medicine. We have developed such an indicator. It is obtained by calculations using Cox's proportional hazard or logistic model with therapy, subgroup, and confounding explanatory variables. The parameter of the interaction between therapy and subgroup can be estimated and tested statistically. The exponential value of the interaction parameter is what we tentatively call the "hazard ratio ratio", meaning the ratio between the treatment effects in two subgroups. The 95% confidence interval of the indicator can also be calculated. As a numerical example, the hazard ratio between the survival times of postoperative gastric cancer patients treated by adjuvant immunochemotherapy and patients without adjuvant immunochemotherapy in a subgroup with high serum glycosidically bound sialic acid (SA) level was lower than that in a low-SA subgroup using an estimate for hazard ratio ratio of less than 0.5 with statistical significance. We propose this indicator be used as a "responder/non-responder ratio" of therapy effect. PMID- 7489570 TI - [The founding and 50-year history of Hradci Kralove Medical School of Charles University--or "Praesente aegroto effugiat risus, tacent colloquia et omnia dominatur mors"]. PMID- 7489569 TI - Tumor labeling in vivo using cyanine-conjugated monoclonal antibodies. AB - Far-red-emitting cyanine fluorochromes have many properties desirable for in vivo imaging: absorption and emission at wavelengths where blood and tissue are relatively transparent, high quantum yields, and good solubility even at high molar ratios of fluorochrome to antibody. Potentially, conjugation by multiple linkages should minimize hydrolysis in vivo. We conjugated two tumor-targeting monoclonal antibodies: anti-SSEA-1 (IgM, kappa) at ratios of 1.2-35 mol dye/mol antibody and 9.2.27 (IgG2a, kappa) at 0.6-6 mol dye/mol antibody, using the cyanine fluorochromes Cy3.18, Cy5.18, and Cy5.5.18. Nude mice were inoculated using the SSEA-1-expressing MH-15 teratocarcinoma or the 9.2.27 antigen expressing SK-MEL-2 melanoma to give tumors at several sites. Conjugated antibody was injected, and mice were imaged immediately after injection and at appropriate intervals thereafter using a standard camera lens, dissecting microscope, or endoscopes. Images were acquired using either an image-intensified video camera or cooled CCD cameras. Immediately after injection, major blood vessels and the heart, liver, and kidneys were readily visualized. After 1 day, tumor-targeting antibody conjugates were concentrated in tumors and there was little circulating conjugate; however, the bladder and kidneys were still visible. Tumors labeled by specific antibody were the most fluorescent tissues at 2 days after injection, but non-specific antibody conjugates did not concentrate in the tumors. The small intestine was weakly visualized by both specific and non-specific antibody conjugates. These data support the possibility of visualizing tumor metastasis by optical means, including currently available endoscopes. PMID- 7489571 TI - [Endocrinology 1994-1995]. AB - In a brief review of advances made in endocrinology during the past two years surprisingly the receptors for TSH and thyroid hormones occupy the foremost position. Mutations of the TSH receptor may cause congenital hyperthyroidism or resistance to TSH (mutation of the gene for TSH-receptor). An interaction of thyroid hormone receptor with a conserved transcriptional mediator was discovered. The reports on the treatment of neonatal respiratory distress syndrome with TRH are still controversial. CRH deficiency reveals neonatal but not adult glucocorticoid need--at least in mice. The concept of receptors was challenged by the discovery of "inverse agonists", i.e. ligand binding by (already) activated receptors. To make endocrinology even more complicated it was shown, that the congenital lipoid adrenal hyperplasia may be caused not by a deficiency in an adrenal cortex enzyme, but by a mutation of the mitochondrial transport protein (StAR), steroidogenic acute protein). The gene for X-linked congenital adrenal hypoplasia (DAX-1) was isolated, its mutation can cause a hypogonadotropic hypogonadism, too. Animals lacking functional estrogen receptor develop osteoporosis--this is true even in humans. Nitric oxide (NO) has more jobs in the body than previously thought-e.g. for maintenance of diurnal rhythms. The interest in endogenous cannabinoids is still flourishing, especially in the brain. Clinical implications of basic research are best documented by the discoveries of G-protein roles in clinical syndromes (e.g. pseudohypoparathyroidism, McCune-Albright syndrome, acromegaly and "testitoxicosis"). It should be emphasized that Nobel prize for medicine and physiology for the year 1994 was awarded to A. G. Gilman and M. Rodbell for the discovery of G-proteins. PMID- 7489574 TI - [Personal experience with surgery of retrosternal goiter]. AB - BACKGROUND: Retrosternal goitre has its specific features: a clinical picture with elements of compression, an increased percentage of malignant conversion, a technically more pretentious operation with an increased number of complications etc. Data in the literature are not uniform. The objective of the presented work was to assemble information in a large group of operated patients. METHODS AND RESULTS: The authors evaluated retrospectively operations of retrosternal goitres from two clinical departments. Of 716 patients operated on account of goitre 87 (21.1%) had retrosternal goitre. Women were affected more frequently (84%) than men (16%), ratio 5.2:1; the mean age of women was 66 years, of men 56 years. A malignant finding was confirmed in 4.6% of the operated patients. The most frequent finding was benign adenomatous colloid goitre (8.5%) and Hashimoto's thyroiditis (80.4%). Hyperthyroidism of the Graves-Basedow type (4.6%) and benign hyperthyroidism of the Plummer type (2.3%) were not frequent. Type of malignity: papillary carcinoma (3.45%) and follicular carcinoma (1.15%); medullary carcinoma, anaplastic carcinoma or lymphoma were not found. The most frequent surgical approach was cervical incision (Kocher) in 95.4%; it had to be supplemented by sternotomy only in 4.6% of the operated patients. As far as the extent of surgery is concerned, almost total thyroidectomy was performed in 60.9%, total lobectomy in 2.3%, subtotal thyroidectomy in 32.2% and subtotal lobectomy in 4.6% of the patients. 24.1% of the patients developed complications, only in 5.7% they were permanent. One female patient died (1.2%). CONCLUSIONS: Retrosternal goitre accounted for cca 20% of patients operated on account of goitre. In cca 5% histology confirmed a malignant form (papillary and follicular carcinoma). Sternotomy was necessary only in less than 5% of the operated patients. The results of surgery are very good. PMID- 7489573 TI - [Tumors of the biliary tract: possibilities of multimodality therapy]. AB - BACKGROUND: Tumours of the biliary system (NBS) account for as many as 16% of all tumours of the GIT. Early diagnosis is difficult, contemporary treatment is not satisfactory and the prognosis is poor. It seems that one of the few therapeutic perspectives is multimodal treatment. The objective of the present work is to assemble experience with multimodal treatment of NBS. METHODS AND RESULTS: From January 1987 to December 1993 347 patients with NBS were treated, using bilioduodenal endoprostheses (BDE). In 26 patients treatment was supplemented by regional chemotherapy, in 10 by intraluminal radiotherapy, using lr 192. Average periods of survival:BDE 230 days, BDE+regional chemotherapy 662 days. Endoprostheses+intraluminal radiotherapy-survival period 96-293 days. The investigation has not yet been completed, as the majority of patients still survives. CONCLUSIONS: Implantation of BDE combined with regional chemotherapy significantly prolongs the life of patients with NBS. It seems that similar results will be obtained also by a combination of BDE and intraluminal radiotherapy. Multimodal treatment of NBS can be nowadays considered the method of choice in the treatment of tumours in this region. PMID- 7489572 TI - [Indicators of lipoprotein metabolism in normolipemic men after myocardial infarct]. AB - BACKGROUND: In some patients with ischaemic heart disease disorders of the lipoprotein metabolism were found although the plasma cholesterol and triglyceride levels were not elevated. The objective of the present work was to assess parameters of the lipoprotein metabolism which could differentiate normolipidaemic probands with an increased risk of myocardial infarction. METHODS AND RESULTS: The authors examined a group of 21 men with normal values of plasma cholesterol and triglycerides who had suffered 6 months to 3 years ago from a myocardial infarction. As compared with a control group of normolipidaemic men without evidence of ischaemic heart disease, these patients had significantly lower HDL-C concentrations (-22%, p < 0.01), in particular due to a lower sub fraction HDL,-C (-29%, p < 0.001), a higher apolipoprotein B concentration in LDL (+36%, p < 0.00) and a lower value of the cholesterol/apolipoprotein B ratio in LDL (-25%, p < 0.05). The total cholesterol, triglyceride, LDL cholesterol, HDL2 cholesterol, apolipoprotein A-1 concentration and cholesterol/apolipoprotein A-1 ratio in HDL did not different significantly. The method of discrimination analysis revealed that for the differentiation of the two groups assessment of apolipoprotein B in LDL and HDL3-C are most useful. By this approach 80% of the patients in the group were correctly classified. CONCLUSIONS: The results of the presented work confirmed the importance of assessment of apolipoprotein B and HDL C, incl. its sub-fractions for estimating the risk of ischaemic heart disease in normolipidaemic subjects. PMID- 7489575 TI - [The human genome--chromosome 18]. AB - The 18th human chromosome is probably known mainly due to patients with Edwards syndrome, children dying usually during the first week of life, the syndrome being due to the presence of three chromosomes 18-trisomy. Some of its loci participate in oncogenesis either as cellular oncogenes (BCL2 in lymphomas), or antioncogenes (DCC--in colorectal carcinomas--Lynch I and Lynch II). It is assumed that some of the causes of maniac depression could be associated with chromosome no. 18. PMID- 7489576 TI - [Metazosin tablets (Kenosin) in the treatment of chronic congestive heart failure]. AB - BACKGROUND: Postsynaptic alpha 1 adrenoceptor blockers are considered suitable drugs for modern vasodilatatory treatment of chronic heart failure. Therefore new analogues of these substances with more favourable characteristics than those of their main representative, prazosin are sought. The objective of the present work was to evaluate the therapeutic effect and tolerance of the original alpha 1 adrenoceptor blocker metazosin developed in the Research Institute for Pharmacy and Biochemistry (VUFB). METHODS AND RESULTS: In a multicentre clinical trial metazosin (tablets a 10 mg), 10-20 mg/day, were added to standard treatment of 56 patients of both sexes suffering from chronic congestive heart failure, for a period of eight weeks. The administration of metazosin led to rapid improvement of subjective complaints (in particular dyspnoea) as well as objective symptoms of peripheral congestion, to a significant (p < 0.05) regression of X-ray signs of pulmonary congestion and a slight (but statistically significant--p < 0.05) increase of the ejection fraction and cardiac output. After two months treatment the functional degree of severity of insufficiency according to the NYHA classification decreased from 2.7 to 2.2. Metazosin did not induce tachycardia and was well tolerated by the patients. CONCLUSIONS: Metazosin (Kenosin) tablets is a suitable supporting drug in the comprehensive treatment of chronic cardiac failure, in particular if refractory to treatment. PMID- 7489577 TI - [Quo vadis pathophysiology?]. PMID- 7489578 TI - [Lies told by the alcohol and tobacco industries and their friends]. PMID- 7489579 TI - [Treatment of anemia in patients with tumors]. AB - About 30% of patients with tumors (in relation to its extent) suffer from anemia which is usually asymptomatic. Etiologically this anemia may be characterized as secondary, so called anemia of chronic diseases. Disorders of iron metabolism, blood marrow insufficiency, extracorpuscular haemolysis, catabolism of patients with tumor burden and relative deficiency of erythropoietin all play a role in its pathogenesis. Anemia of cancer patients may be usually classified as normocytic and normochromic. Indication and timing for treatment of anemia of cancer is equivocal. Successful treatment of anemia seems to improve the quality of life of cancer patients. Indication depends, of course, on the severity of anemia, degree of adaptation and the presence of clinical symptoms related to anemia. Therapy with iron or anabolics is not very effective, therapy with recombinant erythropoietin is not available for all patients, especially for its high price. Transfusion therapy should be considered more carefully in relation with some data showing the possible negative influence of allogeneic blood derivatives on the progression of tumors, especially in patients immunodeficient after high dose chemotherapy and actinotherapy. PMID- 7489581 TI - [Morphologic verification of neoplastic portal vein obstruction]. AB - BACKGROUND: Tumourous invasion into the portal circulation is a frequent and typical complication of primary carcinoma of the liver. No imaging method can, however, assess unequivocally the biological nature of the vascular occlusion and rule out reliably the possibility of benign thrombosis. The objective of the present work was to evaluate the yields of percutaneous thin-needle aspiration biopsy for the verification of intraluminal growth of a tumour, which influences to a considerable extent decisions and possible subsequent therapeutic provisions. METHODS AND RESULTS: Aspiration biopsy of the thrombus was performed in 33 patients with an ultrasonographically apparent occlusion of the portal circulation associated in 29 observations with primary carcinoma of the liver and in four instances with secondaries in the liver. For puncture thin needles--0.8 0.9 mm (21-20 G) were used inserted under ultrasonographic control. The presence of tumourous cells in the portal vessels was proved in 31 patients, i.e. in 94%. An oncologically negative result of biopsy was recorded in one patient with primary carcinoma of the liver and in a female patient with secondaries. Among morphologically verified tumourous occlusions complicating primary carcinoma of the liver a multinodular variant of a tumour was found in 11 observations and a solitary form (mean size of the focus 9 cm) in 10 observations. In 7 patients the vascular occlusion and positive result of biopsy revealed the existence of a diffuse infiltrating type of tumour. In the histological structure of the tumour which was assessed in 13 primary carcinomas of the liver differentiated types of neoplasms predominated unequivocally (10 cases) over non-differentiated tumours (3 cases). CONCLUSIONS: Thin-needle aspiration biopsy is a highly sensitive method for the detection of vascular expansion of liver tumours. It can be used successfully and without risk to assess staging of the disease where necessary. PMID- 7489582 TI - [The effect of pefloxacin on nephrotic syndrome in experimental adriamycin nephropathy]. AB - BACKGROUND: Antiproteinuric effect of pefloxacine was demonstrated in a small group of patients with minimal change nephropathy (MCN) and focal and segmental glomerulosclerosis (FSGS). This finding was not, however, confirmed by other papers. Adriamycine nephropathy is an experimental model of nephrotic syndrome with morphological changes resembling MCN and/or FSGS in patients. METHODS AND RESULTS: Nephrotic syndrome was induced in rats by the i.v. administration of adriamycine. One part of nephrotic animals was treated from the beginning of the 4th week by daily intraperitoneal application of pefloxacine. Administration of adriamycine led in experimental animals after 3 weeks to the development of full blown nephrotic syndrome with further progression of proteinuria in the next 3 weeks (from 1.4 +/- 1.25 to 2.23 +/- 1.89 g of protein/mmol of urinary creatinine, p < 0.05). Proteinuria did not change in nephrotic rats treated by pefloxacine (from 1.04 +/- 0.97 to 1.26 +/- 1.11 g of protein/mmol of urinary creatinine, p = n.s.). The difference in proteinuria between both groups was also significant (0.83 +/- 0.73 vs. 0.23 +/- 0.67 g of protein/mmol of urinary creatinine, p < 0.05). CONCLUSIONS: Pefloxacine was antiproteinuric in experimental adriamycine nephropathy. The mechanism of this effect remains unclear and deserves further studies concentrating on glomerular cytokine network and glomerular production of reactive oxygen species. PMID- 7489583 TI - [The human genome--chromosome 20]. AB - There is an increasing interest on one of the smallest human chromosomes as it is shown by the First International Symposium on the Human Chromosome 20 and by the genetic map prepared by EUROGEN. The conserved part of the long arm of human chromosome 20 is synthenic with the distal part of the mouse chromosome 2 allowing for some analogies between them. Human chromosome 20 contains several important genes for the human pathology. Mutations of one of them, the vasopressin-neurophysin II gene, are responsible for hereditary neurohypophyseal diabetes insipidus. Severe combined immunodeficiency due to adenosin deaminase deficiency is the first human disorder successfully treated by somatic gene therapy. Spongiform encephalopathies are related to mutation and/or polymorphisms of the PRNP amyloid gene. One form of benign familiar neonatal convulsions is mapped to a specific locus on chromosome 20. In some families, maturity onset diabetes of the young (MODY) is caused by alterations of a hypothetical gene closely linked to the ADA locus. Allegile syndrome is often associated with deletions and microdeletions of the short arm of the chromosome. Finally, deletions of the long arm of the chromosome is a frequent finding in several hematologic malignities, specifically in myeloproliferative disorders and myelodysplastic syndromes. PMID- 7489580 TI - [Helicobacter pylori infections: familial clustering and transmission]. AB - BACKGROUND: The importance of Helicobacter pylori for the development of gastritis and gastro-duodenal ulceration is generally accepted. The objective of the presented work was to find out whether its transmission is involved in families of children examined on account of upper dyspeptic syndrome. Aggregation and transmission of Helicobacter pylori infection were not described so far in the Czech Republic. METHODS AND RESULTS: The authors examined IgG antibodies (ELISA) against Helicobacter pylori in 411 subjects incl. 102 children with non ulcerative or ulcerative dyspepsia and in 115 children of similar age but without gastrointestinal complaints and in 194 family members of children with dyspepsia. The specificity of the ELISA test is 97.1% and the sensitivity 92.1% (discrimination analysis). Children with dyspepsia had more frequently a positive finding of IgG antibodies against H. pylori (p = 0.0001), as compared with the control group of children of the same age but without dyspepsia. A statistically significant source of infection with H. pylori for dyspeptic children was the mother (p = 0.0006) and both parents (p = 0.0069), who were also a source of infection for other family members (p = 0.0003 and 0.0084). The most frequent positive IgG finding was in fathers (76.7%), but the father was not a statistically significant source of infection for the examined children (p = 0.6120) nor for other members of the family (p = 0.2330). IgG positive mothers had 85.2% positive husbands, IgG negative mothers had 68.4% positive husbands (p = 0.1601). The frequency of IgG antibodies in the mother and father (in couples) was thus statistically independent. However, there was a significant correlation (p = 0.01) of (P/N) IgG antibodies in couples--i.e. father and mother of the examined children. Siblings of IgG positive children had a significantly higher mean value of antibodies (p = 0.05) and were older. CONCLUSIONS: The presented facts provide evidence that in the investigated group the Helicobacter pylori infection is more frequent in children with the upper dyspeptic syndrome than in children of similar age without gastrointestinal complaints. In families of children with dyspepsia the infection aggregates and is transmitted. The mother and both parents are involved in the transmission to children. Nevertheless it is probable that there is also mutual transmission in children and from adults to children outside the family and in another environment. PMID- 7489585 TI - [History of lung cytology in the Czech Republic]. PMID- 7489586 TI - Whose fault is it? Notes on "true" versus "pseudo" laser failure. PMID- 7489584 TI - [Comparison of the effectiveness and tolerance of Aktiferrin, a ferrous sulfate capsule preparation and Tardyferon pills in patients with uncomplicated sideropenic anemia]. AB - BACKGROUND: The most frequent complications of oral administration of medicinal iron are gastrointestinal complaints the incidence of which correlates with the iron content of the preparation. The objective of the present work was to compare the effectiveness and tolerance of two ferrous sulphate preparations, Aktiferrin capsules and Tardyferon dragees which differ as to the elemental iron content. METHODS AND RESULTS: To two groups of patients with sideropenic anaemia selected at random (39 women and 1 men, age 14-61 years, median 28 years) Aktiferrin or Tardyferon was administered. Administration of the preparations which have a more than double different elemental iron content had a comparable effect on the investigated haematological parameters. In the group treated with Akiferrin no GIT intolerance was observed, in the group with Tardyferon it was observed in four patients. CONCLUSIONS: Aktiferrin has a comparable therapeutic effect although it contains 2.5 times less elemental iron, as compared with Tardyferon. PMID- 7489588 TI - In vivo validation of an experimental adaptive quantitative coronary angiography algorithm to circumvent overestimation of small luminal diameters. AB - The reliability of quantitative coronary angiography (QCA) measurements is of fundamental importance for the study and practice of interventional cardiology. In vivo validation results have consistently reported a tendency for QCA systems to overestimate small luminal diameters. Such a systematic error may result in the underestimation of luminal gain during intracoronary procedures and in the underestimation of progression of coronary artery disease during longitudinal studies. We report the in vivo validation results of an experimental adaptive edge-detection algorithm that was developed to reduce overestimation of small luminal diameters by incorporating a dynamic function of variable kernel size of the derivative operator and variable weighting of the first and second derivatives of the brightness profile. The results of the experimental algorithm were compared to those of the conventional parent edge detection algorithm with fixed parameters. Dynamic adjustment of the edge-detection algorithm parameters was found to improve measurements of small (< 0.8-mm) luminal diameters as evidenced by an intercept of +.07 mm for the algorithm with variable weighting compared to +0.21 mm for the parent algorithm with fixed weighting. A slope of < 1 was found for both the parent and experimental algorithms with subsequent underestimation of large luminal diameters. Systematic errors in a QCA system can be identified and corrected by the execution of objective in vivo validation studies and the consequent refinement of edge-detection algorithms. The overestimation of small luminal diameters may be overcome by the incorporation of a dynamic edge-detection algorithm.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7489587 TI - Influence of expanded balloon diameter on Palmaz-Schatz stent recoil. AB - After successful stent implantation, the residual luminal diameter of the stented vessel is usually smaller than the maximal stent-expanded balloon diameter. The goal of this study was to determine whether immediate vessel diameter recoil after Palmaz-Schatz stenting is affected by the final expanding balloon diameter used during stent deployment. Single Palmaz-Schatz balloon expandable stents were successfully placed in 108 stenotic lesions. There were 68 patients with 75 saphenous vein graft (SVG) and 30 patients with 33 native coronary artery lesions, including 26 restenotic and 82 de novo occlusive (> 50% diameter stenosis) lesions. Quantitative coronary angiography was used for the assessment of stent recoil, defined as the difference between the minimal diameter of the fully expanded balloon and the postprocedure minimal lumen diameter divided by minimal diameter of the fully expanded balloon. A strong correlation (r = 0.94) was found between the minimal diameter of the fully expanded balloon and poststenting minimal lumen diameter. Immediate recoil was 11.3 +/- 7.5%, responsible on an average for 0.4 +/- 0.2-mm acute lumen loss. Recoil was less in SVG than in coronary arteries (9.7 +/- 6.6% vs. 14.0 +/- 7.8%; P = 0.004, and 0.3 +/- 0.2 vs. 0.4 +/- 0.2 mm; p = 0.01). Lesions were divided into four subgroups, based on the final stent expanding balloon diameter: (1) < or = 3.0 mm (n = 33); (2) > 3 < or = 3.5 mm (n = 43); (3) > 3.5 < or = 4 mm (n = 23); and (4) > 4 mm (n = 9).(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7489589 TI - Blood flow velocity alterations during coronary vasospasm. PMID- 7489590 TI - Laser for optical thrombolysis and facilitation of balloon angioplasty in acute myocardial infarction following failed pharmacologic thrombolysis. AB - This report discusses a new indication for the use of holmium:YAG laser. It includes facilitation of thrombolysis and plaque ablation in acute myocardial infarction after failure of thrombolytic agents. Further study is required to define optimal utilization and integration of this novel device for patients who fail to respond to thrombolytic agents. PMID- 7489591 TI - Right coronary artery pseudo-transection due to mechanical straightening during coronary angioplasty. AB - Straightening of tortuous coronary arteries during coronary angioplasty may produce transient angiographic defects that have a typical invaginated appearance. We report a case in which coronary straightening produced an unusual defect mimicking severe damage to the artery. Recognition and conservative management were essential to avoid complications. PMID- 7489592 TI - Left ventricular false aneurysm: an unusually prolonged natural history. AB - A case of left ventricular false aneurysm of postinfarctual etiology with an unusually prolonged natural history (12 years survival) is reported. A first diagnosis of this rare cardiac complication was made in 1982 on the basis of hemodynamic and echocardiographic results. At that time the patient rejected surgical therapy. Eleven years later the patient came back to our attention after resuscitation from a sudden cardiac death. Hemodynamic and echocardiographic (transesophageal) tests showed a remarkable impairment of left ventricular function and an abnormal enlargement (10 x 8 cm) of a pseudoaneurysmatic cavity full of thrombi. The patient died suddenly in April 1994. We emphasize the unusual, prolonged survival of our patient suffering from an unresected left ventricular false aneurysm of postinfarctual etiology. PMID- 7489593 TI - Bacteremia associated with percutaneous transluminal coronary angioplasty. AB - Bacteremia after diagnostic cardiac catheterization is uncommon, but bacteremia after percutaneous transluminal coronary angioplasty (PTCA) has not been studied prospectively. Unlike diagnostic cardiac catheterization, PTCA involves the use of an indwelling arterial sheath after completion of the procedure, which is connected to a pressurized heparin solution, both of which increase the risk of local infection and/or bacteremia. During a 16-week period, we prospectively evaluated patients undergoing 164 PTCA procedures in order to determine the frequency of bacteremia and the significance of fever in this patient population. Blood cultures were obtained from the femoral catheter at the conclusion of the procedure and again 30 min later from the indwelling arterial sheath. Temperature was recorded every 30 min for 2 h following PTCA, then every 4 h over the subsequent 36-hr period. Bacterial isolates were recovered from 23/286 blood cultures (8.0%), with Staphylococcus epidermidis the most common organism present (74%). Only one isolate of Staphylococcus aureus was considered to represent true bacteremia and corresponded with the only documented infectious complication. Fever, defined as > or = 101 degrees F developed in four (2.4%) patients but was procedure related in only one case. The use of the ipsilateral femoral artery for repeat procedures was not associated with either positive blood cultures or difference in maximum temperature elevation. We conclude the overall risk of bacteremia after PTCA is low; therefore, antimicrobial prophylaxis is not warranted. PMID- 7489594 TI - Rotational atherectomy in restenotic lesions at the distal saphenous vein graft anastomosis. AB - Restenotic lesions at distal saphenous vein graft (SVG) anastomoses have been notoriously difficult to treat with standard angioplasty techniques. We explored the potential of rotational atherectomy in three patients with nonthrombotic, focal restenoses at the SVG touch down. The Rotablator safely and successfully recanalized the lesions, allowing further treatment with dilation or stenting or both. Rotational atherectomy may warrant investigation as a treatment for nonthrombotic restenotic SVG lesions. PMID- 7489595 TI - Guide wire extension may not be essential to pass an over-the-wire balloon catheter. AB - The ability to pass an angioplasty balloon catheter over a regular length free guidewire can be advantageous in certain circumstances. This procedure was performed successfully in 99 consecutive patients without any complications. There was improved tactile sense of the lesion and improved dye delivery without the necessity of resorting to the use of extension wires or magnet exchange device. When performed while observing the required precautions this is a safe procedure. PMID- 7489597 TI - Initial experience with an intracoronary flow support device providing localized drug infusion: the Scimed Dispatch catheter. AB - Two cases are presented illustrating the use of a new intracoronary infusion catheter providing flow support using a unique spiral coil design. Good clinical outcomes were obtained employing 4-hr inflations with localized infusion of urokinase at the site of dissection and extensive clot formation, respectively. This low-profile catheter-mounted device may provide an alternative to stent placement in cases of acute dissection complicated by thrombus formation. PMID- 7489596 TI - Emergency rotational ablation of a calcified left main coronary artery stenosis in a patient with ischemic induced cardiogenic shock. AB - High-speed rotational ablation is ideal in the treatment of chronic calcified atherosclerotic lesions. This report describes an elderly woman with a prior history of coronary artery bypass surgery, who developed cardiogenic shock secondary to occlusion of a saphenous vein graft to the obtuse marginal branch of the circumflex coronary artery. Her left main coronary artery was heavily calcified and almost completely occluded. She was treated successfully by high speed rotational ablation of her left main stenosis. PMID- 7489598 TI - Systolic rightward displacement of the left anterior descending artery: a novel cineangiographic sign for tricuspid regurgitation. AB - A new sign for the detection of tricuspid regurgitation (TR) is described. The systolic displacement of the left anterior descending (LAD) artery during coronary angiography in left anterior oblique (LAO) view was quantitatively calculated in 3 groups of 20 patients each with either TR, mitral stenosis, or normal coronary arteriograms. The mean LAD displacement in the TR group was significantly rightward compared to the other groups. Uniform rightward displacement of the lower two-thirds of the LAD had a 90% sensitivity and 90-95% specificity for the presence of TR. Such displacement is probably the angiographic counterpart of the systolic paradoxical septal displacement demonstrated by echocardiography in patients with right ventricular volume overload. There was a positive correlation between the severity of TR and the magnitude of LAD displacement. Attention to the LAD displacement on LAO view may raise suspicion of TR, and indicate its severity. PMID- 7489599 TI - Percutaneous balloon pericardiotomy: a double-balloon technique. AB - We describe a double-balloon technique for performing a percutaneous balloon pericardiotomy. This technique was employed when the large, single dilation balloon customarily used for this procedure failed to fully inflate across the parietal pericardium. Two smaller balloons were advanced through the same skin tract and simultaneously inflated, thus producing an adequate pericardial window. This double-balloon technique allowed for the more secure anchoring of the balloons across the pericardium and for the delivery of greater dilation pressures. PMID- 7489600 TI - Pilot prospective evaluation of counterpulsation with different intra-aortic balloon volumes on cardiac performance in humans. AB - Smaller intra-aortic balloons (IAB) may minimize peripheral vascular complications. To determine the influence of different IAB volumes on cardiac performance, we studied 20 hemodynamically stable patients on IAB counterpulsation. Variables were measured with either a 40cc or 32cc IAB displacement, at an assist ratio of 1:1 or 1:8: Heart rate, Ao and PA pressures, and Fick cardiac output. By echo-Doppler, the velocity time integral (VTI) across the LV outflow tract, a measure of stroke volume was also calculated. The mean age was 60 yr (range 18-77), height 5'6" (5'2"-5'11"), and body surface area 1.9M2 (1.5-2.3). Results presented as mean values were: [table: see text] *P < 0.005 1:1 vs 1:8 for both 40 and 32cc IAB. CONCLUSION: IAB pumping at 1:1 with either 40 cc or 32 cc volume displacement yields similar degrees of improvement in cardiac performance. A larger cohort is required to determine if smaller balloons may decrease complications without compromising efficacy. PMID- 7489601 TI - Responses of systemic and pulmonary veins to the presence of an intravascular stent in a swine model. AB - The outcome of stent implantation for children with pulmonary venous obstruction has been characterized by late reocclusion associated with a marked vessel neointimal proliferation. The purpose of this study was to compare the responses of the systemic vein and pulmonary vein to the presence of an intravascular stent, using a Yorkshire swine (N = 10) model. Under cardiopulmonary bypass, a single Palmaz stent was placed in the inferior vena cava (IVC) and right lower pulmonary vein (PV) with sacrifice at 4.9-6.1 months. Angiography and hemodynamic data were determined at 1 and 3 months post-stent implant and prior to euthanasia. All stents were found to be patent, with no difference in degree of thrombosis or neointimal formation. No statistical difference was found in the initial and final stent diameter for both inferior vena cava and pulmonary vein stents (PV initial 6.8 +/- 0.9; final 7.1 +/- 0.6) (IVC initial 10.4 +/- 1.2; final 10.4 +/- 1.2). Electron microscopy demonstrated smooth endothelialization of both pulmonary and systemic venous stent devices. No thrombosis was found on gross morphology. The data indicate that there is no intrinsic difference in the response of the pulmonary vein to the presence of a stent device. The clinical experience of restenosis following stent implantation for pulmonary vein stenosis appears to be more related to variables of final stent diameter combined with the marked intrinsic abnormal vessel architecture, as seen with this condition. PMID- 7489602 TI - Percutaneous pericardiotomy using Inoue balloon catheter. PMID- 7489604 TI - Electrocardiographic interpretation (1995): can we do better?--Part II. PMID- 7489603 TI - Identifying coronary stenoses prone to thrombosis. PMID- 7489605 TI - Advanced cardiac life support: update on recent guidelines and a look at the future. AB - The objectives of this article are to provide an update of the American Heart Association (AHA) 1992 National Conference guidelines for cardiopulmonary resuscitation (CPR) and emergency cardiac care and to review the investigation and development of new methods of CPR which may be considered in future recommendations. Despite an organized approach to sudden cardiac arrest, survival in patients receiving CPR is in the range of 5-15%. The new AHA guidelines recommend standard manual CPR performed at a rate of 80-100 compressions/min and organized algorithms of advanced cardiac life support. These guidelines stress widespread community training and rapid response in the following sequence: (1) recognition of early warning signs, (2) activation of the emergency medical system (EMS), (3) basic CPR, (4) early defibrillation, (5) intubation, and (6) intravenous medication. Several new recommendations pertain specifically to in hospital care and are, therefore, particularly relevant to physician management of cardiac arrest. The best predictor of survival in patients requiring circulatory support after cardiac arrest is attainable coronary and cerebral perfusion. Unfortunately, the minimal levels of end-organ perfusion required to sustain life are often difficult or impossible to achieve with standard manual cardiopulmonary resuscitation and several new techniques have therefore been introduced. The most promising of these techniques are (1) interposed abdominal compression, (2) pneumatic vest, and (3) active compression-decompression resuscitation. Each of these techniques offers unique advantages when compared with standard manual cardiopulmonary resuscitation. The 1992 National Conference recommendations provide a rational framework for the resuscitation of cardiac arrest victims. New methods of cardiopulmonary resuscitation are now available and investigation into these methods continues.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7489606 TI - Postinfarction ventricular aneurysms. AB - Ventricular aneurysms are circumscribed, thin-walled fibrous, noncontractile outpouchings of the ventricle. The majority are apically located, true aneurysms of the left ventricle (LV) that occur as a consequence of transmural myocardial infarction (MI). The precursor of aneurysm formation appears to be infarct expansion early after acute MI and occurrence generally relates to infarct size. The presence of underlying hypertension and the use of steroids and nonsteroidal antiinflammatory agents may promote aneurysm formation. The clinical sequelae include congestive heart failure (CHF), thromboembolism, angina pectoris, and ventricular tachyarrhythmias. Late rupture is a particular complication of false aneurysms in which the pericardium is the aneurysm wall. The diagnosis may be suspected by the clinical finding of a diffuse, pansystolic apical thrust, persistent ST-segment elevation on the electrocardiogram, and distortion of the cardiac silhouette on chest x-ray. This can be confirmed using echocardiography, radionuclide ventriculography, and cardiac catheterization. The latter has the additional advantage of being able to delineate the coronary anatomy. Management involves prevention, specific therapy for the various clinical manifestations, and surgery. Therapeutic interventions with thrombolytic agents, aspirin, heparin, and beta blockers that are applied early in the evolution of an MI may limit infarction size, thereby reducing the tendency toward infarct expansion and aneurysm formation. Patients with mild CHF can usually be controlled with the standard combination of angiotensin-converting enzyme inhibitors, diuretics, and digoxin. Thromboembolism is best prevented by anticoagulation with warfarin for at least 3 months after the acute MI. The choice of pharmacotherapy for ventricular tachyarrhythmias should be guided by electrophysiologic studies. The treatment of patients with angina pectoris utilizes conventional therapeutic modalities.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7489607 TI - Quantification of mitral regurgitation--comparison of the proximal flow convergence method and the jet area method. AB - A total of 92 patients with mitral regurgitation (age 63 +/- 13 years, 51 men, 41 women), quantified by angiography, were studied using color-flow Doppler imaging of isovelocity surface areas in the flow convergence region proximal to the regurgitant orifice (PISAs) and of the regurgitant jet in the left atrium. The PISA radii for the flow velocities (aliasing borders) of 28 and 41 cm/s, jet area, jet length, and relation of jet area to left atrial area were measured. A proximal flow convergence region was imaged in 98% (85%) of all patients for a flow velocity of 28 (41) cm/s. A regurgitant jet could be visualized in all patients. The PISA radii for both flow velocities correlated more closely with the angiographic grade (rSp = 0.79 for both flow velocities) than the jet area (rSp = 0.43), jet length (rSp = 0.39), and relation of jet area to left atrial area (rSp = 0.37). A correct differentiation of grade I-II from grade III-IV mitral regurgitation was provided in 95% of the patients by the proximal flow convergence method for both flow velocities and in up to 78% of the patients by the jet area method using the uncorrected jet area. The PISA radii correlated weakly with the parameters from the regurgitant jet (r = 0.5-0.58). It can be concluded that the proximal flow convergence method and the jet area method reach comparable sensitivity for the detection of mitral regurgitation.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7489608 TI - Left atrial size--a risk factor for left atrial thrombi in mitral stenosis. AB - In a follow-up study, 77 patients with predominant mitral stenosis were examined to investigate the role of left atrial (LA) enlargement in LA thrombi. Fifteen (19.4%) patients had LA thrombi. Of these, 2 (13.3%) were in sinus rhythm and 13 (86.7%) in atrial fibrillation. Fourteen (93.3%) of the patients with LA thrombi had an LA size > or = 4.8 cm. Only one (6.7%) patient had an LA size of 4.4 cm and was in atrial fibrillation. The median LA size in patients with LA thrombi was 5.2 cm compared with 4.75 cm in patients without LA thrombi (p < 0.01). The relative risk for LA thrombi in patients with an LA size > or = 4.8 cm compared with patients with an LA size < 4.8 cm was 10.0 (95% confidence interval 1.4 to 71.4). It was thus confirmed that LA enlargement > or = 4.8 cm is an independent risk factor for LA thrombi in patients with mitral stenosis. PMID- 7489609 TI - Tilt table evaluation for control pediatric patients: comparison with symptomatic patients. AB - This study was designed to evaluate pediatric control patients during head-up tilt in comparison with symptomatic neurocardiogenic syncope patient head-up tilt responses. Twenty-three pediatric control (c) patients (13 females, 10 males; 11.9 +/- 3.1 years) were tested with head-up tilt (HUT) and compared with 66 symptomatic (s) patients. Baseline drug-free HUT (cHUT-1), a second drug-free HUT (cHUT-2), and a final HUT with isoproterenol infusion (cHUT-3) were each performed at 80 degrees tilt angle for 30 min or until positive. For comparison, 66 symptomatic patients (41 females, 25 males; 13.6 +/- 2.5 years) underwent drug free HUT (sHUT-1); negative responders during sHUT-1 underwent follow-up HUT with isoproterenol (sHUT-2). HUT data were compared for both groups at both 30 and 20 min tilt duration. Twelve control patients (52%) had a symptomatic response during cHUT-1 at 18 +/- 8 min. During cHUT-2, 5 of 23 patients were positive at 13 +/- 5 min; each had previously tested positive during cHUT-1. Two patients, each positive in cHUT-1 and cHUT-2, refused cHUT-3. The only patient testing positive during cHUT-3 was test positive in cHUT-1 but negative for cHUT-2. In comparison, 43 of 66 (65%) symptomatic patients tested positive during drug-free sHUT-1 at 11 +/- 6 min. Subsequently, 20 of the 23 negative patients underwent HUT with isoproterenol (sHUT-2), with 8 of 20 testing positive. Thus, 51 of 66 symptomatic patients (77%) were called "true positives."(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7489610 TI - Lipoprotein (a) is increased in acute coronary syndromes (unstable angina pectoris and myocardial infarction), but it is not predictive of the severity of coronary lesions. AB - Lipoprotein (a) [Lp(a)] concentrations were determined in 365 patients undergoing coronary angiography for stable angina (n = 159), unstable angina (n = 99), recent myocardial infarction (n = 45), and nonischemic heart disease (cardiomyopathy or valvular disease, n = 62, non-IHD). Mean +/- SD and median Lp(a) concentrations in stable angina (29.9 +/- 29.2;22 mg/dl) did not differ from those in non-IHD (26.9 +/- 26.3; 17), but were significantly lower than in patients with unstable angina (52.7 +/- 36.6; 58) and myocardial infarction (44.8 +/- 36.4; 34) (p < 0.01). Coronary angiography revealed that 261 patients, including 4 patients in the non-IHD group, had significant (> or = 50%) coronary lesions. Lp(a) was higher in patients with (41 +/- 35; 32) than in those without (28 +/- 27; 19) angiographic evidence of significant coronary stenosis (p < 0.05) and showed a weak univariate correlation with the angiographic index (Total Score) of the severity of the disease (r = 0.106;p < 0.05). However, in the subgroup of 303 patients with stable/unstable angina or myocardial infarction, Lp(a) was predictive neither of angiographic presence nor of severity of coronary disease. Patients were then ranked according to the Total Score values. Among patients with comparable angiographic severity of coronary artery disease, Lp(a) appeared to be remarkably higher in patients with acute ischemic syndromes (unstable angina, myocardial infarction) than in patients with stable angina. In conclusion, Lp(a) was roughly twice as high in acute (unstable angina, myocardial infarction) than in chronic (stable angina) ischemic syndromes, but there was no difference between chronic stable angina and non-IHD.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7489611 TI - Intracardiac thrombi: frequency, location, etiology, and complications: a morphologic review--Part II. AB - Intracardiac thrombus is a frequent finding at necropsy. Various cardiac disorders have been associated with its presence. Part II of this 5-part article focuses on atherosclerotic coronary heart disease (myocardial infarction) as an etiology for intracardiac thrombus. PMID- 7489612 TI - The effect of body position on a free-floating ball thrombus as observed by transesophageal echocardiography. AB - Two cases of free-floating left atrial ball thrombi (FLABT) in association with mitral stenosis were observed by transesophageal echocardiography (TEE). Our report describes the relation between body position and thrombi kinetics. Both cases demonstrated similar kinetics. In the supine and right lateral decubitus positions, the thrombi recoiled from and sometimes became entrapped within the mitral valve. In the sitting and left lateral decubitus positions, the thrombi appeared to be nearly fixed and did not contact with the mitral valve. Our results indicate that the latter two positions prevent thrombi disintegration and incarceration into the mitral valve. Finally, TEE is an extremely useful tool for assessing the safest position for individuals with FLABT. PMID- 7489613 TI - Left ventricular outflow tract pulmonary artery fistula in endocarditis. AB - Patients with infective endocarditis are at risk for the development of a fistulous communication between chambers or great vessels of the heart. The presence of a continuous murmur may suggest the diagnosis. The first case of aortic valve endocarditis complicated by the development of a fistulous communication between the left ventricular outflow tract and the pulmonary artery is reported. Transesophageal Doppler echocardiography did not detect the defect preoperatively. However, pulmonary artery catheterization revealed very high mixed venous oxygen saturation which supported the presence of a left-to-right shunt. PMID- 7489614 TI - Louis Faugeres Bishop. PMID- 7489615 TI - Molecular genetics: cardiac disease and risk-related genes. AB - Molecular genetics is playing an increasing role in the diagnosis, treatment, and prevention of cardiac disease. Moreover, most of the genes that may cause cardiac disease or predispose an individual to cardiac disease are anticipated to be identified within the next 10 years. Several genes with risk for heart disease have been identified, such as the ACE genotype DD. Replacement gene therapy as well as use of promoter-specific drugs to act on genetic regulatory elements will encompass the future treatment of cardiovascular disease. This article provides a summary of the potential roles of genetic screening for cardiac risk factors and genetic interventions in cardiovascular disease. PMID- 7489616 TI - Modulating cardiac growth: a new paradigm in the treatment of heart failure. Introduction. PMID- 7489617 TI - Tumor necrosis factor-alpha and the failing human heart. AB - Tumor necrosis factor-alpha (TNF alpha) is a proinflammatory cytokine with negative inotropic effects. Recently, elevated levels of TNF alpha have been identified in patients with advanced heart failure. Although the clinical significance of this finding is unclear at present, there is increasing evidence that this cytokine may play a primary pathophysiologic role in the development and pathogenesis of heart failure in humans. Indeed, many of the clinical hallmarks of heart failure, including left ventricular dysfunction, cardiomyopathy, and pulmonary edema can be explained by the known biological effects of TNF alpha in humans. The present review will summarize recent evidence with regard to the biological role for TNF alpha in the adult mammalian heart, as well as summarize the increasing body of clinical information that implicates this cytokine in the pathophysiology of heart failure. PMID- 7489619 TI - Cell death in the failing heart: role of an unnatural growth response to overload. AB - Hypertrophy of the overloaded heart, characterized by an increased number of sarcomeres, provides an adaptive, short-term response. However, when cardiac overload is long-standing, the hypertrophic response appears to cause shortened myocyte survival. The mechanisms responsible for the deleterious effects of chronic myocardial hypertrophy may include a maladaptive growth response of the mature heart. Because terminally differentiated adult cardiac myocytes have little or no capacity to divide, stimuli that promote growth in the overloaded adult heart cannot lead to normal cell division. Instead, overload initiates an unnatural growth response that appears to shorten cardiac myocyte survival, possibly because the same growth factors that mediate the hypertrophic response of the adult heart can also induce programmed cell death (apoptosis). The converting enzyme inhibitors and nitrates, which have growth-inhibitory as well as vasodilator effects, may improve prognosis in heart failure by inhibiting the production of transcription factors. These transcription factors stimulate both the unnatural growth response to overload and stimuli that lead to apoptosis. Since both beta-adrenergic agonists and cytokines, such as tumor necrosis factor alpha, can stimulate production of similar transcription factors, evidence suggests that beta blockers and vesnarinone improve the prognosis in patients with heart failure possibly because of their ability to inhibit maladaptive growth. PMID- 7489618 TI - Strategies for pharmacologic modulation of the heart failure phenotype. AB - The end-stage heart failure phenotype is characterized by marked dyspnea on exertion, edema, and overwhelming fatigue, and by a high incidence of sudden death. Patients who display the end-stage phenotype have transitioned from a normal phenotype with myocardial damage at a cellular level. This transition appears to be mediated by events at both the cellular and molecular levels. Until recently, it was generally believed that this transition was irreversible. However, recent clinical trials have demonstrated that the phenotype can be changed with pharmacologic agents. These agents have been demonstrated to improve exercise capability, increase ventricular function, and improve symptoms. Important recent studies have shown that pharmacologic agents can substantially alter the high mortality rates associated with the end-stage heart failure phenotype. As we learn more about the molecular and cellular events that initiate and support the transition from cardiac compensation to decompensation, we will be able to improve our pharmacologic targeting and, we hope, be able to delay the development of the end-stage heart failure phenotype to an even greater degree. PMID- 7489620 TI - Critical review of heart failure: the role of left ventricular remodeling in the therapeutic response. AB - Recent clinical investigations have provided new insights into physiologic alterations taking place as heart failure progresses. As a result, new end points may be more relevant to monitor in patients, rather than the classical assessment of severity based on subjective evaluation of functional capacity. Structural changes of the myocardium, referred to as remodeling, have profound effects on the performance of the left ventricle and on long-term prognosis. Left ventricular ejection fraction may serve as a clinical marker for remodeling changes. Remodeling changes involve hypertrophy of the muscle mass, dilation of the left ventricle, and a change in the shape of the ventricle. Histologically, these changes are produced by myocyte lengthening, interstitial growth, and myocyte slippage. The consequences of remodeling include an abnormal bio energetic state because of the augmented wall stress and accentuated myocardial oxygen consumption. The elevated wall stress promotes further hypertrophy. Superimposed on the remodeling changes is neurohormonal activation. Both remodeling and neurohormonal activation are directly related to mortality. An experimental canine model has been used successfully to investigate the potential impact of pharmacologic intervention on remodeling. Angiotensin-converting enzyme inhibitors and the combination of hydralazine and isosorbide dinitrate have been shown in clinical trials to reduce long-term mortality and increase ejection fraction. The effects of these drugs appear to be mediated in part by alterations in remodeling and neurohormonal activation. Thus, left ventricular ejection fraction can be used as a clinical marker for remodeling processes, and plasma norepinephrine levels can be used as a marker for neurohormonal activation. PMID- 7489621 TI - Magnetic resonance angiography of the distal lower extremity. AB - OBJECTIVE: To assess magnetic resonance angiography (MRA) for demonstration of arterial patency in the ankle and foot of patients with peripheral vascular disease. METHODS: Peripheral MRA of the ankle and foot was performed on 34 limbs of 31 insulin-dependent diabetics. 2-D time-of-flight MRA (TR 33 ms/TE 7.7 ms/inferior saturation band) was performed with 16 cm field of view. Pre- or intra-operative angiographic correlation was available in all cases. RESULTS: In 24 limbs MRA was compared to conventional angiography. MRA showed more patient run-off vessel segments (120) than angiography (100). In 10 limbs MRA was compared to intraoperative angiography and for the detection of patent vessel segments showed a sensitivity of 87.5% (42/48) with a 95% confidence interval of 75% to 95% and a specificity of 95% (38/40) with a 95% confidence interval of 83% to 99%. Pitfalls included difficulty in visualizing flow at the bifurcation of the peroneal artery, in the plantar arch and retrograde flow in the lateral plantar artery. CONCLUSIONS: MRA is sensitive for the detection of patent arteries in the ankle and foot but artefacts may cause overdiagnosis of focal stenoses or occlusions. PMID- 7489622 TI - Accelerated thrombolysis with high dose bolus t-PA extends the role of peripheral thrombolysis but may increase the risks. AB - Low dose intra-arterial thrombolysis is too slow for many patients with severe acute limb ischaemia. Accelerated thrombolysis with high dose bolus t-PA was used in a consecutive series of 43 patients. Complete or clinically useful lysis was achieved in 39 patients, with a median duration of 7 h. Lysis occurred in 46% in under 4 h. Fifty-six per cent of patients required further procedures after lysis. Eleven per cent suffered a major bleed. The limb salvage rate at 30 days was 56%. Amputation was required in 22% and 22% died. Most deaths were due to associated thrombotic conditions: myocardial infarction (5), pulmonary embolism (1) and malignant thrombosis (1). One patient died from pneumonia two weeks after lysis and two died from renal failure within a week of thrombolysis. The high mortality rate was not associated with bleeding but may reflect the high risks involved in treating this group of patients. High dose bolus t-PA infusion appears to predict immediate outcome of thrombolysis as well as reducing infusion times. It may expand the indications for the non-surgical treatment of acute limb ischaemia to include most patients with the condition. Careful case selection is still necessary for optimal results. PMID- 7489623 TI - Pseudothrombosis of the infra-renal inferior vena cava during helical CT. AB - Pseudothrombosis of the infra-renal vena cava is a frequent finding during post contrast helical computed tomography (CT) studies of the abdomen. Three hundred consecutive patients undergoing contrast-enhanced helical studies were evaluated prospectively to document the incidence of pseudothrombosis. Typical CT findings are illustrated and a significant difference in incidence between the male and female population noted. PMID- 7489624 TI - Cryptococcal pneumonia in patients with the acquired immunodeficiency syndrome. AB - Infection with Cryptococcus neoformans occurs with increased frequency in patients with the human immunodeficiency virus (HIV). Despite the lungs being the portal of entry for the fungus the commonest presentation is with meningitis: Cryptococcal broncho pulmonary infection occurs less commonly. The chest radiographs of fourteen HIV positive patients with cryptococcal pneumonia were reviewed. The commonest radiographic abnormalities were interstitial infiltrates, (nine patients) and focal or widespread alveolar consolidation (seven patients). Ground glass shadowing, not previously described in cryptococcal pneumonia, occurred in six radiographs, miliary nodules in one, lymphadenopathy in four, and small pleural effusions in three. Two patients had a normal chest radiograph. In contrast to previous reports, we found alveolar consolidation and ground glass shadowing to be common and nodules and cavitation to be rare. Cryptococcal pneumonia should be considered in the differential diagnosis of Pneumocystis carinii pneumonia, bacterial pneumonia and miliary tuberculosis in HIV positive patients. PMID- 7489626 TI - The CT small bowel faeces sign: description and clinical significance. AB - OBJECTIVE: To describe a new CT sign of the GI tract: the small bowel faeces sign, and discuss its significance. METHODS: The small bowel faeces sign consists of gas bubbles mixed with particulate matter in dilated segments of small bowel. This was a retrospective study of 22 patients demonstrating this sign from 1989 to 1993. Final diagnosis was established by surgical, medical or laboratory findings. To determine the prevalence of the sign, the CT examination of 120 consecutive separate control patients were evaluated. RESULTS: All 22 of the patients demonstrating this sign required hospitalization with surgical or medical intervention. Eighteen of 22 had mechanical small bowel obstruction. The remaining four patients had other abnormalities of small bowel to account for the finding on CT. Twelve of the 22 patients were treated with surgery and the remaining 10 patients were treated with nasogastric tubes (n = 6) or other medical therapy (n = 4). None of the 120 control patients demonstrated the sign. CONCLUSION: The presence of gas and particulate material resembling faeces in a dilated segment of small bowel on CT is abnormal. Most (18/22; 82%) patients with this sign had small bowel obstruction. PMID- 7489625 TI - A sonographic-pathological correlation of liver biopsies in patients with the acquired immune deficiency syndrome (AIDS). AB - The liver is commonly involved in patients with AIDS and a first line investigation for hepatic dysfunction is liver ultrasound (US) which is often abnormal. It is unclear how these US abnormalities correlate with the underlying pathological processes. A retrospective study was performed in 48 patients with HIV disease who had undergone both liver biopsy and hepatic (US), correlating the findings. Only 25% of patients had an entirely normal liver (US) examination and only 10% patients had a normal liver biopsy. The commonest sonographic abnormalities was a diffusely hyperechoic liver seen in 46% of patients and this correlated with steatosis. Forty-four percent of biopsies contained more than one histological abnormality including granulomas, inflammation, steatosis or siderosis. In addition clinically unsuspected pathology was revealed in five of the cases of mycobacterial infection and in three cases of lymphoma. The combination of multiple histological abnormalities, unsuspected pathology, and the altered immune response in this group makes the US findings even less specific than in non-AIDS patients. We recommend that liver biopsy should remain an essential and early part of the management of AIDS patients. PMID- 7489627 TI - Ultrasound in the differential diagnosis of appendicitis. AB - The purpose of this study was to determine the accuracy of sonography in detecting diseases other than appendicitis in the clinical setting of suspected appendicitis. Six hundred and nine patients were evaluated using graded compression ultrasound (US). A broad spectrum of diseases was suggested in the group of patients who eventually turned out not to have appendicitis (n = 426), including gastrointestinal (154), gynaecological (32), biliary (11), urological (8), and miscellaneous (5) abnormalities. However, out of these there were three cases of incorrect sonographic diagnoses with serious sequelae. Appendicitis coincidental with another disease capable in itself of explaining the patients' symptoms was confirmed in seven out of 10 patients where sonographically suggested; in six patients out of this group only the suggested alternative condition was held responsible for the symptoms while the inflamed appendix was overlooked. US is of considerable value in establishing alternative diagnoses in patients with equivocal signs of appendicitis. However, one should always consider the possibility of co-existing appendicitis in patients where US has revealed another condition which could be held in itself responsible for the patient's symptoms. PMID- 7489628 TI - Classifying interval cancers. AB - No definitive way of classifying interval breast cancers (cancers presenting between screening rounds) has been determined, yet their number and classification forms one of the quality assurance (QA) standards for the National Health Service Breast Screening Programme (NHSBSP). This study was undertaken to identify how different screening centres undertake this process, and to compare the classifications obtained when a test set of mammograms was reviewed using three different methods. A questionnaire was sent to the 17 Regional UK breast screening QA centres. Twelve (80%) of the 15 centres completing the questionnaire had a formal method for reviewing interval cancers. Five of these (33%) attempted to simulate screening by mixing the interval cancers with other screening films. In 11 (73%) centres, a group (size range 3-14) of radiologists was involved. In a simulated film viewing exercise we assessed whether different methods of classification would alter the number of interval cancers classified as false negative (where an abnormality suspicious of malignancy can be identified on review of the original screening films). Six radiologists reviewed a set of 50 interval cancers by three different methods: independent reading of the interval cancers mixed with screening mammograms; independent reading of the interval cancers on their own; and finally a consensus opinion of the interval cancers alone using the films taken at diagnosis. No discussion or review of these cases had taken place prior to the study. The number of interval cancers classified as false negative increased by 10% when they were reviewed in isolation compared to review among 'normal' screening films. The false negative rate varied widely (4% to 56% P < 0.01) depending on the criteria required to fulfil a false negative interval cancer classification--whether only one, a majority, or all of the radiologists were required to see the abnormality on the original screening films. In order for inter-unit and regional comparisons to be useful, a standard review method of mixing the interval cancer films with normal screening films and using a consensus opinion by a minimum of three external reviewers is suggested. PMID- 7489629 TI - Relative incidence and morphology of lateral and medial meniscal cysts detected by magnetic resonance imaging. AB - It is likely that meniscal cysts of the knee are a complication of meniscal tears which extend to the capsular attachment. They are reported as occurring three times more commonly on the lateral than the medial side. We reviewed 50 cases of meniscal cysts diagnosed on magnetic resonance imaging in patients with suspected internal derangement of the knee. Twenty-eight cysts involved the medial and 22 the lateral meniscus. The most common sites were the posterior third of the medial meniscus and the mid-third of the lateral meniscus. Fifteen cysts were suspected clinically, five on the medial side and 10 on the lateral side. Small cysts were only clinically predicted on the lateral side. We conclude that menial cysts are more common than previously thought and are more difficult than lateral cysts to detect clinically. PMID- 7489630 TI - Inflammatory pseudotumours in children: CT and ultrasound appearances with histopathological correlation. AB - Inflammatory pseudotumour is an uncommon benign lesion that presents in children and young adults. The rarity of these lesions, particularly at extrapulmonary sites, has resulted in poor documentation of its radiological manifestations. The casenotes, radiology and histology of five patients with inflammatory pseudotumour were reviewed. Two lesions were intra-abdominal, one oesophageal, one intrapulmonary and one lower limb. CT demonstrated inflammatory pseudotumours as well circumscribed masses of soft tissue density producing displacement of surrounding structures rather than local invasion. Sonography depicted these lesions as well defined masses with homogeneous echo patterns. Surgical removal resulted in dramatic symptomatic improvement. PMID- 7489632 TI - The radiological treatment of hepatic artery aneurysms. AB - Improvements in the quality and availability of cross-sectional imaging should result in more frequent detection of hepatic artery aneurysms before rupture. Interventional radiological treatment for extra-hepatic lesions has not previously been discussed in the literature. We present two cases of extra hepatic hepatic artery aneurysms treated using different endovascular techniques and discuss the relative merits of these approaches. PMID- 7489631 TI - Traumatic bone cysts in the jaws of a Hong Kong Chinese population. AB - The trabecular bone cyst (TBC) is the jaws' equivalent of the unicameral cyst of the long bones. A series of 20 TBCs were surgically treated in the jaws (19 in the mandible and one in the maxilla) of 14 Chinese patients (five males and nine females), who could broadly be divided into two age-sex groups; a young group of both sexes and an older, exclusively female group. Three patients in the older group had more than one TBC and the one with initially five separate TBCs (including one in the maxilla) exhibited six episodes of recurrence of the mandibular lesions which finally coalesced to affect the whole body of the mandible bilaterally. PMID- 7489633 TI - Technical report: kinking of a Simon nitinol vena caval filter during insertion. PMID- 7489634 TI - Case report: granular cell myoblastoma of the breast: a rare benign tumour mimicking breast carcinoma. PMID- 7489635 TI - Case report: periosteal neurofibroma mimicking osteochondroma. PMID- 7489636 TI - Case report: intracranial calcification following herpes simplex encephalitis. PMID- 7489637 TI - Case report: percutaneous transluminal angioplasty of irradiation induced bilateral subclavian artery occlusions. PMID- 7489638 TI - Case report: percutaneous transhepatic portographic embolization (PTPE) for recurrent hepatic encephalopathy derived from intrahepatic portal-systemic shunts. PMID- 7489639 TI - Interventional radiology of the pleural space. PMID- 7489640 TI - Biologic treatment of human cancer. AB - Biologic therapy of cancer is defined as treatment that produces antitumor effects through the action of natural host immune mechanisms or the administration of natural animal substances. Biologic therapy is considered to be a potentially important fourth modality for the treatment of cancer alongside surgery, radiation therapy, and chemotherapy. Increased application of this form of cancer treatment will result from a better understanding of the basic aspects of host defense mechanisms against cancer and significant technologic developments that have made molecules available in quantities large enough for use in human patients. Although the field of biologic therapy remains relatively early in its development, there are many in vitro and in vivo examples of the successful application of biologic therapy to human cancer. Moreover, several recombinant biologic therapeutic agents in current use are considered standard therapy for selected malignant diseases. Future research in this field will focus on improved modulation of the human immune system and is anticipated to improve the current therapeutic armamentarium used against cancer. PMID- 7489641 TI - [Three dimensional analysis of the facial lateral region with beautiful appearance of Chinese and its clinical value]. AB - 100 Chinese young men and women with attractive appearance and normal occlusion were the subject of this study. The shape and constructive characteristics of the masseter region and mandibular angles were measured and analyzed by Moire photography. A serial parameters were obtained, which are important in the correction of the deformities of facial asymmetry. A typical case with bilateral hypertrophy of masseters and mandibular angles was reported. The surgery was directed by the results of preoperative Moire photography analysis. PMID- 7489642 TI - [Tolerance of reconstructive flaps to postoperative irradiation in oral and maxillofacial regions]. AB - Eighty-three reconstructive flaps in oral and maxillofacial regions irradiated with a dose of 4000 to 7200 cGy were observed for a 2 year follow-up period. It was found that the rates of acute side effects including redswelling erosion and ulceration in the flaps were lower significantly than in neighbor oral mucosa (P < 0.01-0.05). The late radiation side effects of the flaps were uncommon also. The result showed that free flaps were well tolerated to postoperative irradiation of routine dose. Our studies indicate that the preoperative irradiation deficiency of blood supply or purulency were main reasons of the late radiation side effects, e.g. atrophy of epithelia, cicatrization or necrosis of the flap. Base on the results we have designed preventive measures. PMID- 7489643 TI - [The diagnosis and treatment of the epithelial salivary gland tumors in children and adolescents]. AB - Sixty-six cases with epithelial salivary gland tumors in children and adolescents, who were less than 16 years old, were analized. Those cases accounted for 3.6% of all epithelial salivary gland tumors in our school. Twenty five cases (37.9%) were benign tumors, among which pleomorphic adenoma was the most common. Forty-one cases (62.1%) were malignant tumors, in which mucoepidermoid carcinoma was the most common one. Three quarters of cases were in the group over 10 years old. Six cases were in the group less than 5 years old. Among them 5 cases were malignant tumors. The younger the children is, the more possible the malignant tumor is. Two cases with malignant tumors had the history of receiving radiotherapy because of the hemangiomas at the same locations. It indicates that the occurrence of the malignant tumors was related to radiotherapy before. The diagnosis, differential diagnosis and treatment of the tumors were discussed. The initial radical resection of the malignant tumors was emphasized. PMID- 7489645 TI - [A preliminary study on the mechanism of against human tongue cancer cell line]. AB - Inverted phase contrast microscope and transmission electron microscope examination reveals that, tumor-draining lymphnode lymphocytes (DNL) conjugated with tumor cells very tightly at 12 hours co-culture. The DNL cell nucleus was seen positioned away from the target cells, whereas the cell mitochondria granules were oriented toward the point of contact with the target cells. The injured target cell had a highly condensed cytoplasm and chromatin and formation of cytoplasmic blebs, which were consistent with an apoptotic cell death rather than with a lytic or necrotic cell death. The mechanism was exocytosis of granules containing a pore-forming protein. Holes formed in the target cell's membrane may allow a lethal substance to enter and to induce its death via an apoptotic mechanism. PMID- 7489644 TI - [Light microscope and energy dispersive X-ray analysis of amalgam pigmentation]. AB - 24 cases of amalgam pigmentation were analyzed by light microscope. 16 cases were amalgam debris, 8 cases were amalgam fragments and debris mixed. The amalgam distributed mainly along basement membrane of the epithelium, wall of blood vessels, nerve fibers, reticular fibers and endomysiums. In case where amalgam entered the bone, bone cells disappeared, lacuna emptied and microfracture of bony trabecullae occurred. In 8 cases pathologic calcification were found and in 18 cases there were inflammatory responses. 2 cases were analyzed by energy dispersive X-ray. It was found that in the different sites of tissues there were different content of Ag, Hg, Sn and Ca. PMID- 7489646 TI - [Observation on variations of cyclic nucleotide content in human oral-maxillo facial cancers]. PMID- 7489647 TI - [The ultrastructure study of white sponge nevus in oral mucosa]. AB - Biopsies from 7 patients with white sponge nevus were examined histologically. Two of them had family history. Light microscopic observation reveals that the epithelium was thickened; the spinous cell layers had enlarged and oedematous cells and ballon-like degeneration; the keratohyalin granules were increased in upper spinous cells; the superficial layers of epithelium exhibited parakeratosis and thickening. 3 specimens were examined by electron microscopy. The basal and parabasal cells were cylindrical. The cytoplasmic tonofilaments were aggregated into boundles. It was characteristics of keratinized epithelium. In middle and superficial stratum spinosum the tonofilaments were homogeneously distributed in the cell plasma and the electron-dense granulars were located near the cell boundary and attached to desmosomes by tonofilaments, unlike normal non keratinized oral mucosa. Many Odland bodies with a lamellar structure and irregular in both size and shape were seen. It is concluded that this leasion have a dyskeratinized process. The characteristic features of keratinized oral mucosa in non-keratinized mucosa is appeared. The thickening of superficial layer is due to shedding disturbance relating to dysfunction of Odland bodies. PMID- 7489648 TI - [Effect of dentin organic and inorganic substances on the curing time of bonding resin]. PMID- 7489649 TI - [An immunohistochemical study of HLA-DR expression in salivary glands from patients with Sjogren's syndrome]. AB - Specimens of salivary gland tissue from 21 cases of SS were analyzed by ABC immunohistochemistry technicue with a monoclonal antibody to HLA-DR. The results showed that positive immunostaining for HLA-DR were found in duct cells, acinar cells and a part of infiltrating cells respectively. The proportions of duct and acinar cells expressing HLA-DR were not increased between SS and controls. However, there are differences in their staining intensity. The HLA-DR expression on the epithelial cells in SS was most intense than in controls. Our data suggest that the HLA-DR expression of SS may be regarded as an important factor in local immune rections. PMID- 7489650 TI - [Chronic suppurative parotitis: proposed classification]. PMID- 7489651 TI - [Experimental study of promoting the rates of dog's tooth rotation by circumferential supracrestal fiberotomy just before rotation]. PMID- 7489652 TI - [The complication and treatment of 233 cases of radical neck dissection]. PMID- 7489653 TI - [Clinical studies of preoperative tumor reductive chemotherapy with PPV regimen in the treatment of oral squamous cell carcinoma]. PMID- 7489654 TI - [Relation of wedge-shaped tooth defects, the quality of toothbrush and toothbrushing methods]. PMID- 7489655 TI - [Titanium porous-surface screw implant]. AB - Titanium porous-surfaced screw implant was made by plasma-spray with Ti powder. The pore size was 80-300 microns. Two groups of implants were implanted in the mandibles of 15 dogs. Group A: porous-surfaced screw implant: Group B: smooth surface screw implant. The bone healing was evaluated by L. M. SEM. and shear strength testing. The results show that the osteo integration of two groups were completed after 12 weeks. The bone grew into porous surface. The shear strength of group A was significantly higher than that of group B. PMID- 7489656 TI - [Clinical studies of a caries-defense varnish containing sialin-like material]. PMID- 7489658 TI - [Miniplate rigid fixation in orthognathic surgery]. PMID- 7489657 TI - [Preparation of monoclonal antibodies against 185-SA of Streptococcus mutans]. PMID- 7489659 TI - Investigation of EEG non-linearity in dementia and Parkinson's disease. AB - Many recent studies based on the surrogate data method failed to identify significant non-linearity in the EEG. In this study we examine whether the use of a different embedding method (spatial instead of time delay), and calculation of Kolmogorov entropy (K2) and the largest Lyapunov exponent (L1) in addition to the correlation dimension (D2), can distinguish the EEG form linearly filtered noise. We have calculated D2, L1 and K2 of original EEG epochs and surrogate (phase randomized) data in 9 control subjects, 9 demented patients and 13 Parkinson patients. The correlation dimension D2 and the largest Lyapunov exponent L1 could distinguish between the EEG tracings and the surrogate data. Demented patients had significantly lower D2 and L1 compared to controls. L1 was higher in Parkinson patients than in demented patients. Contrary to other studies that have used the Theiler surrogate data method, we find evidence for non-linearity in normal and abnormal EEG during the awake/eyes closed state. Apparently it is the spatial structure in the EEG that exhibits much of the non-linear structure. Furthermore, non-linear EEG measures show more or less specific patterns of dysfunction in dementia and Parkinson's disease. PMID- 7489660 TI - Slow wave electroencephalic activity parallels renin oscillations during sleep in humans. AB - Previous studies have demonstrated that the nocturnal oscillations of plasma renin activity (PRA) exactly reflect rapid eye movement (REM) non-REM (NREM) sleep alternation with levels of PRA that increase during NREM sleep and decrease during REM sleep. These studies were based exclusively on conventional scoring of sleep stages. In the present study, we used spectral analysis of the sleep EEG to determine the variations in the different EEG frequency bands, together with PRA profiles. Eight male volunteers participated in a 1 night study. They were subjected to 8 h polysomnography including spectral analysis of the EEG, and to blood sampling every 10 min. Delta relative power and Sleep Intensity Index and PRA oscillations ran parallel in all individuals. An increase in slow waves was associated with an increase in PRA, whereas a decrease was associated with a decrease in PRA. Cross-correlation coefficients were significant and ranged between 0.34 and 0.74. Conversely, theta, alpha and beta bands and the EEG mean frequency were inversely proportional to PRA, with lower cross-correlation coefficients. These results may give further support to the hypothesis of a common mechanism controlling both SWA and renin release from the kidney. PMID- 7489661 TI - Movement-related potentials associated with bilateral simultaneous and unilateral movements recorded from human supplementary motor area. AB - To clarify the differences of movement-related potentials (MRPs) among ipsilateral, contralateral and simultaneous bilateral movements, MRPs with finger, thumb or foot movements were recorded from subdural electrodes chronically implanted on the supplementary motor area (SMA) in 3 patients, and also from the primary sensorimotor area in two of them being evaluated for epilepsy surgery. As a result: (1) SMA generated clear pre-movement potentials regardless of the type of movement. Its amplitude was almost identical between contralateral and bilateral movements except for the motor potential (MP). The pre-movement potentials associated with ipsilateral movements were relatively smaller than those with contralateral or bilateral movements. (2) The primary sensorimotor area generated clear pre-movement potentials in contralateral and bilateral movements with similar amplitude. With ipsilateral hand movements, however, only a small Bereitschaftspotential (BP) and no negative slope (NS') or MP was seen, and ipsilateral foot movements were not preceded by any BP. It is, therefore, most likely that, as far as the preparation for simple voluntary self paced movement is concerned, the SMA plays an equally important role in unilateral and bilateral movements, whereas the primary sensorimotor area is involved predominantly in the preparation of contralateral movements. PMID- 7489662 TI - Movement-related cortical potentials associated with voluntary muscle relaxation. AB - We recorded movement-related cortical potentials (MRCPs) associated with voluntary muscle relaxation, which was not accompanied by contraction of the antagonist or any other muscles, in 10 normal subjects. Voluntary, self-paced relaxation of the wrist extensors from the extended position was employed as the relaxation task, and wrist flexion by muscle contraction was employed as the contraction task. The accelerogram was used to obtain the trigger signals for both tasks. The electromyograms of the ipsilateral agonist and antagonist, the proximal muscles and the contralateral corresponding muscles were monitored to confirm the absence of muscle contraction for the relaxation task. All MRCP components were identified in both tasks; Bereitschaftspotential (BP), negative slope (NS'), parietal peak of motor potential (ppMP) and frontal peak of motor potential (fpMP). BP started earlier and was larger at the contralateral parietotemporal electrodes for the relaxation than for the contraction task, and the slow positive shift at the bilateral frontopolar electrodes was seen more often in the relaxation task. It is concluded that the voluntary muscle relaxation needs a cortical preparatory process similar to voluntary muscle contraction, and needs a more extensive and longer preparation process in the primary motor area and possibly in other motor areas as compared with the contraction. PMID- 7489663 TI - Contingent negative variation and movement-related cortical potentials in parkinsonism. AB - The contingent negative variation (CNV) and movement-related cortical potentials (MRCPs) were recorded in 10 cases of Parkinson's disease, 10 cases of vascular parkinsonism and 10 age-matched healthy controls. The amplitude of the early CNV was smaller in the Parkinson's disease group and the vascular parkinsonism group than in the healthy control group. The latencies of Bereitschaftspotential (BP) and negative slope (NS') were longer and the amplitude of BP was smaller in the Parkinson's disease group and the vascular parkinsonism group than in the healthy control group. These abnormalities in CNV and MRCPs were improved by intravenous injection of L-DOPA. PMID- 7489664 TI - Movement-related cortical potentials in persistent mirror movements. AB - Mirror movements (MMs) are involuntary movements executed on one side of the body during voluntary movements of the contralateral homologous body parts which may abnormally persist into adulthood. In 6 subjects affected by persistent MM with autosomal dominant inheritance, movement-related cortical potentials (MRCPs) during self-paced, voluntary extensions of either the left or right middle finger were recorded from 30 EEG electrodes simultaneously with the electromyogram (EMG) of both extensor digitorum communis muscles. The negative potentials before and during EMG onset were evaluated statistically for the two electrodes next to the cortical hand areas. A comparison with 7 normal subjects revealed no marked differences for the Bereitschaftspotential (BP) and the negative slope (NS'). Only in the periods around EMG onset (from -50 to +50 msec) a significant difference between both groups was found. The MM subjects showed fairly symmetric potentials over the right and left hemispheres, whereas the potentials of the control subjects were lateralized to the hemisphere contralateral to the intended movement. No difference was found for the amplitude of the maximum negative peak of MRCP following EMG onset. Our data showed no evidence for a different type of movement preparation in MM subjects as compared to normals. We propose that the additional ipsilateral cortical activation around movement onset may be the cortical mechanism, which compensates for abnormal ipsilateral corticospinal pathways in subjects with persistent MM. PMID- 7489665 TI - Pharmacologically induced changes in arousal: effects on behavioral and electrophysiologic measures of alertness and attention. AB - The relationships between the diffuse subcortical neurotransmitter systems and behavioral and physiologic measures of alertness and attention are not well understood. This study was designed to further understand these relationships. In this double-blind experiment, 23 subjects ingested methylphenidate, diphenhydramine or placebo on 3 different days and performed behavioral and cognitive tasks including covert orienting of spatial attention and visual search tasks. Subjective and physiologic measures of alertness included EEG frequency analysis, EEG event-related desynchronization, and amount of sleep and sleep onset time in the unstimulated eyes closed state. Performance on the cognitive tasks improved with MP and worsened with DPHA, but there were no specific attentional effects. The best measures of alertness were based on self-rated scales and on EEG recorded in the unstimulated eyes closed state. These observations suggest that methylphenidate and diphenhydramine primarily affected overall state and that healthy humans were able to partially compensate for the pharmacologically induced alertness changes during cognitive task performance. PMID- 7489666 TI - Comparison of Z and multivariate statistical brain electromagnetic maps for the localization of brain lesions. AB - Conventional Z maps provide probability statements about the deviation of observed values from the norm. Galan et al. (1994) introduced Simultaneous Significance Probability Scales to detect abnormalities over the whole map, making use of the information provided by the topographic structure of dependencies. They also described multivariate brain electromagnetic (MBE) maps for compact presentation of complex spatio-temporal information. In this paper, using the distance-based localization receiver operating characteristic curves (DL-ROC curves), we compare the localization provided by computed tomography with that provided by Z and MBE maps in 61 patients with brain lesions. Maps were calculated for absolute power and relative power in delta, theta, alpha and beta bands for voltage and current source densities (CSD). In each patient, all maps were compared and the map with the highest value of the area of the DL-ROC curve was considered to be the "best map." Z maps of CSD were the "best maps" in 24 patients. In the voltage montage, we observed that multivariate maps add some information not contained in the Z maps. However, for CSD, Z maps were more accurate than multivariate maps. A very consistent finding was the observation that lesions were better detected by maps analyzing the delta band, while edema was better represented by maps in the theta range. PMID- 7489667 TI - Improved forward EEG calculations using local mesh refinement of realistic head geometries. AB - A method for semi-automatically constructing realistic surface meshes of 3 head structures--scalp, skull and brain--from a stack of MR images is described. Then an evaluation is given for both spherical and realistic dipolar models, using the boundary element method (BEM). In both cases, locally refined models were considered. Two characteristic mesh parameters were defined: the global and the local mesh densities (in triangles per cm2). In spherical geometries, numerical and analytical solutions were compared, and in the realistic case, all models were compared to a highly refined one, considered as a reference. Both geometries gave comparable results. It was found that for "deep dipoles" located at more than 20-30 mm under the brain surface, meshes with a global density of 0.5 tri/cm2 gave "acceptable" results, whereas for more superficial dipoles (2-3 mm < depth < 20-30 mm), it was necessary to locally refine meshes near the source location up to a local density of about 5-8 tri/cm2, to get comparable results. PMID- 7489668 TI - Dynamic spectral analysis of event-related EEG data. AB - A method for analysing the time course of power spectra of event-related EEG data is presented. A sequence of autoregressive models is fitted to segments of the EEG within which the data exhibit local stationarity. For parameter estimation a method involving ensemble averages is introduced. Besides investigating the evolution of power spectra, time courses of peak frequency, bandwidth and power of alpha (mu) and beta rhythms are traced. The method is applied to EEG recorded over the primary motor area during self-paced finger movements. PMID- 7489669 TI - Clinical evaluation of VEPs to interleaved checkerboard reversal stimulation of central, hemi- and peripheral fields. AB - The VEPs of 195 patients referred for supportive evidence of multiple sclerosis or optic neuritis were studied by a new method of interleaved checkerboard reversal stimulation of different areas of the visual field. In the first group of 95 patients checks of 40' subtense reversed in the whole field (28 degrees x 20 degrees), alternatively in the left and right hemifields and alternately in the central (5 degrees radius) and peripheral fields. In the second group of 100 patients checks reversed in the whole field and in interleaved mode in 3 visual field areas, comprising the central (4 degrees radius) and left and right hemisurround fields. In the first group abnormal responses were recorded from 52 eyes and there was partial disagreement among the stimulus conditions in 10 of the 52. Abnormalities were seen uniquely to central field stimulation in 3 eyes but never to whole field stimulation alone. In the second group abnormal responses were recorded in 58 eyes, again never uniquely to whole field stimulation, while abnormalities confined to one or two areas of the visual field were seen in 24, providing evidence of peripheral field involvement alone in 8 eyes. In the first group, waveforms created from the sum of the left and right hemifield and central and peripheral field responses showed quite close conformity to the whole field VEP, although amplitudes were significantly lower and latencies significantly shorter. In 7 eyes responses would have been differently classified (normal or abnormal) using the sum as compared with the whole fields.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7489670 TI - Electrophysiological exploration of visual function in mitochondrial diseases. AB - In a group of 10 children (ranging from 5 months to 15 years old) affected by diseases with mitochondrial dysfunction, 4 suffered from mitochondrial myopathy, 4 from mitochondrial encephalopathy and 2 from Friedreich's ataxia. The clinically detectable visual impairment consisted of 3 grey ocular fundi (the other 7 were normal) associated, in 2 subjects, with a mild nystagmus. Electrophysiological assessment, consisting of ERGs and flash VEPs, was systematically performed. The normal ERGs in all subjects confirmed the normal functioning of retinal electrogenesis. In contrast, the VEPs of 6 out of 10 subjects were modified: in 2 of the 4 subjects with mitochondrial myopathy, the VEPs had a hyperamplitude; in the 2 subjects with Friedreich's ataxia, the implicit time of the principal VEP peaks was increased, together with a hyperamplitude in 1 case; lastly, in 2 of the 4 subjects with mitochondrial encephalopathy, the VEPs were altered. These modifications reflected visual pathway conduction disorders with no clinical expression. Various underlying pathophysiological mechanisms possibly responsible for these modifications are discussed. PMID- 7489671 TI - Short latency visual evoked potentials to flashes from light-emitting diodes. AB - Short latency visual evoked potentials (SVEPs) have been described in response to high-intensity, strobe flashes. High-intensity flashes can now be generated from goggle-mounted light emitting diodes (LEDs) and the SVEPs to such flashes have been shown to be reproducible across subjects, avoiding photic spread to the examination room and acoustical artifacts from the strobe stimulator. In this study, SVEPs from multichannel records are described in terms of normative latencies and amplitudes, as well as scalp distributions, to explore their generators. Potentials were recorded from 10 young male subjects, from 16 scalp locations, in response to flashes from goggle-mounted LEDs. Flashes were presented to each eye in turn, as well as binocularly. The latencies, scalp distributions and intersubject variabilities of the LED evoked SVEPs were similar to those obtained with strobe flashes. SVEP components were divided into 3 groups, according to their latency and the electrodes at which they were recorded with the largest amplitudes: periocular (under 40 msec latency), fronto-central (40-55 msec) and parieto-occipital (55-80 msec latency). The scalp distributions observed in this study suggest subcortical generators along the visual pathway, beginning at the retina. The use of goggle-mounted LEDs should promote routine evaluation of the integrity of the visual pathway between retina and cortex using SVEPs. PMID- 7489672 TI - Clinical value of the pudendal somatosensory evoked potential. AB - The pudendal evoked potential was recorded in 126 patients who had presented with various uro-genital complaints. The patients were divided into two groups depending on whether or not there were clinical signs of neurological disease. Group I consisted of 83 patients (66%) who on clinical examination were neurologically normal. In group II there were 43 patients (34%) who had physical signs suggesting underlying neurological pathology. The pudendal evoked potential was abnormal in 10 patients from the group with neurological signs (group II) but only 1 patient from group I (a man who had made an excellent recovery from previous transverse myelitis). The conclusion of this study is that the pudendal evoked potential is of no greater value than the clinical examination in the assessment of patients with uro-genital dysfunction. The recommendation that the potential should be recorded as part of the routine assessment of patients suspected of having a neurogenic disorder of the bladder and sexual function should be reconsidered. PMID- 7489673 TI - Modulation of sural nerve somatosensory evoked potentials during stance and different phases of the step-cycle. AB - In order to investigate the modulation of somatosensory processing during stance and locomotion, sural nerve somatosensory evoked potentials were recorded during both stance and different phases of the step-cycle. Characteristic sequences of negative-positive waves were elicited, consisting of an early component, N40, presumably of subcortical origin, followed by a P50-N80-P220 complex of cortical origin. The N40 and N40-P50 components had similar amplitudes in both gait and stance. However, the P50-N80 component was attenuated whereas the N80-P220 complex became biphasic during gait. Within the step-cycle, amplitudes of the cortical components P50-N80 and N80-P110 were larger prior to footfall and smaller at the beginning of the support phase. The results demonstrate that locomotion produces a modulatory effect on somatosensory input at a cortical level. Within the step-cycle, excitability of the somatosensory cortex is increased during the middle and late swing phases and decreased during the support phase. Such modulation may contribute to an improved detection of foot contact at touchdown. PMID- 7489674 TI - Auditory evoked potentials in Down's syndrome. AB - Short-, middle- and long-latency auditory evoked potentials (SAEPs, MAEPs and LAEPs) were examined in 12 subjects with Down's syndrome and in 12 age-matched normal subjects. In comparison with the normal subjects, Down subjects showed shorter latencies for SAEP peaks II, III, IV and V (and correspondingly shorter interpeak intervals I-II and I-III) so long as stimulus intensity was at least 45 dB SL. The MAEP peak Na had a longer latency in Down subjects than in normal subjects, but not the Pa latency. In passive oddball experiments for LAEPs, the latencies of all components from N1 to P3 were progressively longer in Down subjects, and the N2-P3 amplitude increased slightly between the first and fourth blocks of stimuli (whereas in the normal subjects it decreased). These alterations in auditory evoked potentials, which may correlate with cerebral alterations in organization and responsiveness responsible for deficient information processing, may constitute an electrophysiological pattern that is characteristic of Down's syndrome. PMID- 7489675 TI - Electric source localization of the auditory P300 agrees with magnetic source localization. AB - The event-related cortical potential elicited in the context of auditory target detection tasks includes the N1, P2 and P3 components. The aim of the present study was to identify the sources of these scalp-recorded components using an electrical multiple dipole model. Nine healthy adults volunteered for the study. An auditory oddball paradigm was used. Stimuli (18% target and 82% non-target tones) were delivered through ear-phones and subjects were required to silently count the targets. Event-related potentials (ERPs) to these stimuli were recorded by 30 electrodes placed on the scalp. The identification of the sources of the ERP was attempted using the brain electric source analysis (BESA) program. The instantaneous source locations of N1, P2 and P3 reported in magnetoencephalographic (MEG) literature were used as initial starting locations for the spatio-temporal multiple dipole modeling of the EEG data. First the auditory long latency responses were modeled separately. Bilateral superior temporal plane sources with almost vertical orientations explained the first 250 msec window of the non-target tone recording including N1/P2 complex. This agrees with MEG source localization of N1m/P2m. Two slightly deeper dipoles in superior temporal gyri and bilateral dipoles in hippocampi or parahippocampal areas explained P3 (analysis window 250-600 msec). The final model explained the complete epoch of 600 msec with 6 dipoles and the residual variances of individual models ranged from 3.83% to 7.77%. The concordance between MEG and BESA source localization results supports the notion of generators in temporal lobes for the N1/P2 complex and generators in temporal and hippocampal areas for the P3 component. PMID- 7489676 TI - Variability and replicability of the mismatch negativity. AB - The interindividual variation and test-retest stability of the mismatch negativity (MMN) and N1 components of the event-related potential (ERP) were investigated by presenting standard (85%) and deviant tones (15%) to 10 young subjects in 2 sessions separated by 1 month. Deviant tones in different blocks were either frequency or duration changes with interstimulus intervals (ISIs) of 0.5 and 1.5 sec. The results showed a fairly good test-retest stability of the MMN amplitude for both types of changes with each ISI at the group level. The amplitude of the duration MMN showed significant individual test-retest stability. The N1 amplitude showed high stability at both the group and individual levels. Both the MMN and N1 showed considerable interindividual variation. The results suggest that MMN and N1 can be used in follow-up studies not only at the group level but possibly at the individual level also. PMID- 7489677 TI - P3b-like potential of rats recorded in an active discrimination task. AB - We investigated whether the potential corresponding to the human P3b could be recorded on the dura mater over the frontal cortex of the rat in an active discrimination task. Rats were trained to press a bar within 1200 msec after cessation of the target tone (1000 Hz) lasting for 800 msec, and to withhold an overt response to the standard tone (2000 Hz). Rats were given intracranial electrical stimulation to the medial forebrain bundle as a reward only when they correctly responded to the target tone. The stimulus probability of the target tone was manipulated at 3 levels: 30, 50 and 70% of all trials. Large, slow positive deflections with peak latency at about 400-500 msec were elicited to the target tone, which were preceded by a large negative potential with its peak around 180 msec, whereas such a clear deflection was not elicited to the standard tone irrespective of the stimulus probability. This positive slow deflection was very similar in morphology to the human P3, and this P3-like potential was significantly larger when the target tone was less frequent (30%) than in the case of more frequent targets (50 and 70%). These results suggest that a potential highly similar to P3b can be recorded in the rat, and that it may provide a useful model to investigate the neurophysiological basis of the human P3b. PMID- 7489678 TI - The Duffing oscillator: a model for the dynamics of the neuronal groups comprising the transient evoked potential. AB - Thirty years ago Zeeman conjectured that the dynamics of the EEG might be modeled by the "equation of motion" of a Duffing oscillator, a pendulum with a nonlinear, cubic, restoring force. In this study the idea is extended to the evoked potential (EP). A transient sensory EP reflects activity from several neuronal groups, pools of neurons that fire in synchrony, with voltage-time curves that overlap appreciably. When the dynamics of each neuronal group is modeled by a Duffing oscillator, multi-electrode transient VEPs are well predicted. Predictions based on Duffing oscillator dynamics are substantially better than those based on the assumption that each neuronal group follows a simpler exponentially damped sinusoid or a function that simulates a post-synaptic potential. The component voltage-time curves are reasonably consistent over 7 subjects, suggesting sequential activation of neuronal groups with delays of several tens of milliseconds between them. The scalp topographies of the components suggest their origins in the occipital cortex. PMID- 7489679 TI - Signal detection in averaged evoked potentials: Monte Carlo comparison of the sensitivity of different methods. AB - Many clinical and research applications rely on detecting evoked potential (EP) signal or EP differences between conditions. Statistical methods for objective signal detection should be sensitive to the presence of signal, but must provide the user strict control on tolerated false alarm rate. The respective sensitivities of 6 signal detection methods were compared through several Monte Carlo simulations involving 2 autocorrelation structures, 5% and 1% significance levels, 8, 10 or 12 replications per study, and increasing signal to noise ratio. The signal detection methods compared were: (1) the Record Orthogonality Test by Permutations (ROT-p), a variant of the Residual Orthogonality Test (Achim et al., 1988), that provides an unbiased estimate of the energy of the signal present in the averaged data, (2) the Tsum2 permutation test of Karniski et al. (1994), (3) a Principal Component Analysis method (PC1) consisting of a t test on the weights of the first principal component, (4) multiple t tests on amplitudes with empirical adjustment for global false alarm rate, and (5-6) the test of Guthrie and Buchwald (1991) on length of consecutive t tests significant at P < 0.05 or 0.01 per-test. The first 3 methods did not exceed their nominal false alarm rate and clearly outperformed the last 3, with the ROT-p method being significantly more sensitive than all others under almost all conditions. PMID- 7489680 TI - Discriminant classification of motor unit potentials (MUPs) successfully separates neurogenic and myopathic conditions. A comparison of multi- and univariate diagnostical algorithms for MUP analysis. AB - Multivariate statistical methods may be more appropriate for the multidimensional material of quantitative motor unit potential (MUP) analysis than the multiple univariate tests of the conventional Buchthal analysis. Buchthal analysis was slightly modified before it was used as the gold standard for new multivariate diagnostical algorithms, based on principal component analysis and on MUP discriminant classification: muscle means of continuous variables were related to tolerance limits after adequate transformation. Chi-square tests were used for dichotomized variables, e.g., polyphasia. Sensitivity and specificity of the uni- and multivariate algorithms were compared for 539 muscles from patients with motor neuron diseases, neuropathies and myopathies and for 91 biceps brachii, rectus femoris and tibialis anterior control muscles. False positive results accumulated less than expected by repeat univariate tests for single MUP parameters, due to high correlation. Combination of single parameters to factor scores did not improve specificity. One advantage of factor analysis was that factor matrix and factor scores matched those of previous studies in spite of different input parameters, which may facilitate multicenter comparisons. Discriminant classification successfully separated neurogenic and myopathic conditions, even in myositic muscles and motor neuron diseases, where myopathic and neuropathic MUPs frequently intermingle. Discriminant classification may support expert decisions and add weight to EMG differential diagnosis. PMID- 7489681 TI - Normal values of nerve conduction in children and adolescents. AB - Healthy children and adolescents (n = 128) ranging in age from 6 to 20 years and in height from 114 to 193 cm underwent studies of median, peroneal and sural nerves bilaterally, including nerve conduction velocity, amplitude and motor distal latencies. Arms and legs were heated in all subjects to obtain skin temperatures around 34 degrees C. Both motor and sensory nerve conduction velocities were found to correlate more with height than with age. There was a strong negative correlation between height and peroneal conduction velocity (r = 0.40, P < 0.0001). On the contrary, a positive correlation was found between height and both median sensory (r = 0.30, P < 0.0001) and motor (r = 0.22, P < 0.001) conduction velocities. Skin temperature, even near 34 degrees C, had a strong effect on conduction velocity and motor distal latencies. It is concluded that consideration of height and temperature will improve the diagnostic safety of nerve conduction measurements in children and young adults. PMID- 7489682 TI - Parkinson's disease rigidity: EMG in a small hand muscle at "rest". AB - The presence of excessive EMG at "rest" might be an important factor in the genesis of Parkinson's disease (PD) rigidity, and we studied it in the first dorsal interosseous muscle (FDI) of 8 idiopathic PD patients. We had 8 age- and sex-matched normal controls. In the PD group, the average area of the surface EMG at "rest" correlated significantly with the clinical evaluation of rigidity and remained abnormally enhanced for 10-15 min after a command to "relax." Later, it tended to decline, but its entity was still much greater than in controls. The EMG "at rest" consisted of unwilled motor unit (MU) firing. A larger MU number was recruited in patients than in controls at "rest." MU rate coding was similar in both groups. Eventually, patients could get periods of EMG silence which, however, were interrupted by short EMG bursts, even if there was no muscle stretch. These bursts were interpreted as residual fragments of the original excessive EMG at "rest." MUs first recruited during such bursts showed high, but not total, overlapping with those first recruited by a gentle voluntary contraction or by a weak transcranial magnetic stimulus to motor cortex. We conclude that EMG activity at "rest" was made up of the discharge of low threshold MUs, with a recruitment order similar to that resulting from descending cortico-spinal volleys. However, we cannot exclude other possible input sources to the alpha-motoneurones at "rest." PMID- 7489684 TI - Trial-to-trial variability of corticospinal volleys in human subjects. AB - The trial-to-trial variability of the different components of corticospinal volleys evoked by transcranial electrical stimulation of the motor cortex using a constant stimulus intensity was measured from epidural recordings during surgery to correct scoliosis. The recordings were made when there was no operative interference, and blood pressure, temperature, ventilation and anaesthetic regimen were stable. A simple D wave with a single negative peak of 10-30 microV amplitude was recorded in 4 patients. It varied little in amplitude (S.D.s < 8% for 100 consecutive single responses). In 4 patients the stimulus was adjusted to produce a complex D wave with 3 components, the earliest 2 of which arise from subcortical/brain-stem sites. The variability of amplitude of these components was high (S.D.s of 13-50%), but the variability of latency was low (S.D.s of 2 3%). Eighteen I waves were recorded in 6 of the subjects. Their variability from trial to trial was similar to that of the components of the complex D wave. It is argued that there would be greater trial-to-trial variability of the corticospinal volley in the awake state, particularly when the stimulus was magnetic rather than electrical. Explanations for changes in the compound muscle action potential produced by transcranial stimulation, electrical or magnetic, must take into account that a constant stimulus does not evoke an identical descending volley. PMID- 7489683 TI - Effects of premotion silent period on single motor unit firing at initiation of a rapid contraction. AB - We compared the single motor unit (SMU) activity between movements with a premotion silent period (PMSP) and without PMSP in EMG. Fourteen SMUs in the gastrocnemius muscle and 6 SMUs in the soleus muscle were recorded from 5 volunteers during isometric plantar flexion. Tonically firing SMUs failed to fire just before the onset of a rapid contraction with PMSP more frequently than without PMSP. SMUs tended to fire within 10 msec (the gastrocnemius SMUs) or 20 msec (the soleus SMUs) from the onset of the phasic EMG discharge when PMSP occurred. In a rapid contraction without PMSP, the initial firing of SMUs occurred with longer latency than that in a rapid contraction with PMSP. The latency of the initial SMU firing in a rapid contraction related to the preceding time of the last SMU firing during a sustained contraction to the onset of the phasic EMG discharge. When the preceding firing was long enough, the latency distributed around 10 msec. On the other hand, for shorter preceding times, the latency lengthened with shortening of the preceding time. It is suggested that the PMSP makes the preceding time long and increases the susceptibility of motor units to the descending command at the initiation of a rapid contraction. PMID- 7489685 TI - The boundary effect in magnetic stimulation. Analysis at the peripheral nerve. AB - The optimal stimulus position for a figure-8-shaped coil for magnetic stimulation of the ulnar nerve at the wrist was not coincident with the optimal electrical stimulus point but was shifted 18.3 mm to the ulnar side (P < 0.01). For the median nerve the optimal stimulus site was 9.6 mm radial to the optimal position for electrical stimulation (P < 0.05). This shift of the stimulus point for magnetic stimulation is significantly smaller after interposition of a homogenous electrically conducting medium between coil and arm but not changed after interposition of distilled water. This so-called boundary effect is therefore due to the different conductivities of the medium interposed between coil and nerve. It may also distort precise localisation of other excitable structures such as cranial nerves, nerve roots and cortical areas by means of magnetic stimuli. The amplitudes of the compound muscle action potentials elicited with identical magnetic stimulus strength were larger after the interposition of isotonic solution between coil and skin but not after interposition of distilled water. Consideration of the boundary effect provided an improved response amplitude to magnetic stimulation, but this could not adequately compensate for its poor localisation compared to electrical stimulation. PMID- 7489686 TI - Comparison of the magnetically mapped corticomotor representation of a muscle at rest and during low-level voluntary contraction. AB - This study has compared the topography of the corticomotor representation of an intrinsic hand muscle in the relaxed state and during a low-level voluntary contraction using transcranial magnetic stimulation. It was found that the optimum cortical stimulus site and the surrounding area of excitable cortex were shifted approximately 6 mm medially as a result of performing a voluntary contraction. This difference may be due to a combination of factors involving the spatial distribution of excitable intracortical fibres, the effects of muscle afferent activity on motor cortex excitability, and task-dependent modifications of corticomotor output. PMID- 7489687 TI - Ipsilateral motor response--is it an artefact? AB - The lack of information about the ipsilateral motor response to cortical stimulation prompted the present study. Motor evoked potentials (MEPs) in 60 patients with acute stroke were recorded bilaterally. The patients' ages ranged between 24 and 83 years; 45 were males. Intracerebral haemorrhage was present in 23 and infarction in 37 patients. In none of these patients was an ipsilateral response recorded from the hemiplegic side when a contralateral response was absent. On the non-hemiplegic side in all these patients an MEP was recorded on both contralateral and ipsilateral cortical stimulation. Unrecordable MEPs became recordable in 6 patients after 1-15 months of stroke. In these patients MEP latencies on ipsilateral cortical stimulation were either identical to that on contralateral stimulation (2 patients) or within the range of normal intraindividual variation. On the basis of these results, it is concluded that the ipsilateral motor response in our study may be due to cross stimulation of the normal hemisphere and not through the uncrossed motor pathways. Our results should be interpreted in the context of the stimulation conditions employed. Further studies are recommended using more focal stimulation to minimise the spread of current. PMID- 7489688 TI - Prognostic significance of central motor conduction on the non-hemiplegic side in supratentorial hematoma. AB - Central motor conduction abnormalities have been reported on the hemiplegic side only. In a study on supratentorial intracerebral hemorrhage (ICH), we found central motor conduction time (CMCT) prolongation on the non-hemiplegic side in 12 patients. This study reports the prognostic significance of CMCT prolongation on the non-hemiplegic side. CT scans were reviewed and CMCT was measured at the 2nd, 4th and 12th week. Recovery was defined on the basis of the 3 months' Barthel index score. Their mean age was 52 (range 25-60) years, 6 were males. Eight had putaminal, 3 thalamic and 1 frontal hematomata. The size of the hematoma was large in 5 and moderate in 7. The clinical signs of herniation were present in 5; radiological evidence of midline shift in 10 and tentorial herniation in 6 patients was also present. There was no other CT abnormality to account for the prolongation of CMCT on the non-hemiplegic side which returned to normal in all 5 patients after a mean duration of 34 (range 11-120) days. Five of these patients died and the remaining had a poor outcome. We conclude that CMCT prolongation on the non-hemiplegic side in supratentorial hematoma suggests a poor prognosis. PMID- 7489689 TI - Motor cortex changes in a patient with hemicerebellectomy. AB - To evaluate reorganisation of motor pathways following a cerebellar lesion, we studied motor cortex excitatory responses and inhibitory effects after transcranial stimulation, together with segmental spinal cord excitability, in one patient who had undergone hemicerebellectomy. We compared the results obtained using different forms of stimulation capable of activating the cortico spinal tract at different sites. Results were compared between sides. We previously reported that the threshold for responses is higher in the motor cortex contralateral to the impaired hemicerebellum and the right/left threshold asymmetry is clearly greater than normal when a circular coil centred over the vertex is used. In the present study, using electrical anodal stimulation, no side difference was observed. Significant interside differences were absent also when the durations of the silent periods or the mean amplitude of the flexor carpi radialis H reflex between the two sides were compared. The outcome is that the interside differences previously observed are mainly due to reduction in the intrinsic excitability properties of the motor cortex functionally related to the impaired hemicerebellum and not to modification of the inhibitory properties of the cortex or to spinal mechanisms. PMID- 7489690 TI - Inhibitory phenomena in individual motor units induced by transcranial magnetic stimulation. AB - It is well known that a silent period (SP) can be observed in voluntary tonic EMG activity starting directly after the initial early response when magnetic stimuli are delivered through the skull over the contralateral primary motor cortex. It is, however, unknown as to how an individual motoneurone (MN) contributes to the SP observed in the surface EMG. The present investigation was conducted to investigate inhibitory phenomena at the level of individual motor units. It demonstrates that the duration of the SP in single motor units is inhomogeneously distributed within the pool of active MNs. At various stimulation strengths, SP durations in single motor units can be similar or longer when compared to that observable in surface EMG records. In some motor units, which show low thresholds for early excitation and appearance of the SP, durations of SP can exceed 1000 msec. The length of suppression of spontaneous MN firing is maximal at stimulus intensities a little higher than those required for an early excitatory response. Although in general thresholds for early excitation and appearance of SPs are similar, at threshold stimulation in a number of trials inhibitory effects on the firing of voluntarily activated motoneurones were present, even in the absence of early excitations. This proves the independent nature of inhibitory as opposed to excitatory effects induced by transcranial magnetic stimulation. An SP in the absence of early excitation underlines its cortical origin. Inhibition and excitation of single MNs were maximal over the same small scalp area. We suggest that cortical inhibitory control plays an important role in the organization of natural movements. PMID- 7489691 TI - A stable late soleus EMG response elicited by cortical stimulation during voluntary ankle dorsiflexion. AB - Transcranial electrical or magnetic stimulation was performed in 47 experiments on 42 normal adult subjects. Surface compound muscle action potentials were recorded from the antagonistic tibialis anterior (TA) and soleus (SOL) muscles. A stable late response recorded from SOL during voluntary ankle dorsiflexion is described and called soleus MEP-80. Its origin has been studied with different experimental procedures. (1) It is only obtained during active voluntary dorsiflexion of the ankle within 70-100 msec (mean 86.9 +/- 6.4 msec) whereas passive dorsiflexion does not promote such a response. The size of SOL MEP-80 was always bigger than the primary SOL response. (2) A similar constant response has not been obtained during voluntary plantar flexion from SOL and TA muscles. (3) SOL MEP-80 is not changed significantly by the Jendrassik manoeuvre. (4) It is not obtained or is suppressed when the subject is sitting with the feet free in space. On the other hand, it is obtained when the subject is standing, especially when standing is made difficult. (5) SOL MEP-80 has not been obtained by brain stem, spinal cord or root stimulation. (6) A later response of longer than 100 msec latency was sometimes obtained at rest but such an S100 soleus response was abolished or transformed to the SOL MEP-80 immediately during ankle dorsiflexion. (7) It is concluded that SOL MEP-80 is a polysynaptic extensor response related to postural mechanisms and originating through convergence of descending motor commands and peripheral sensory feedback. PMID- 7489692 TI - Developmental aspects of seizure semiology: problems in identifying localized onset seizures in infants and children. PMID- 7489693 TI - Complex partial seizures of temporal lobe origin in children of different age groups. AB - The semiology of complex partial seizures (CPS) of temporal lobe origin in adults is well known and is important in establishing seizure localization in patients considered for epilepsy surgery. In contrast, the behavioral features of temporal lobe seizures (TLS) in children described in the literature have not been consistent. In the present study, we investigated children with TLS to compare their attacks to TLS occurring in adults. The study was based on video recordings of 29 children with TLS aged 18 months to 16 years. Children were included, if they became seizure-free after temporal lobectomy (except 4 children with a marked reduction in seizure frequency and 1 with isolated auras), and if clear unitemporal seizure onset in ictal EEG-recordings, unilateral radiological lesions, and corresponding histopathological findings were detected. Children aged > 6 years had TLS with features similar to those of adults. In younger children, typical semiology included symmetric motor phenomena of the limbs, postures similar to frontal lobe seizures in adults, and head nodding as in infantile spasms. We concluded that the clinical features of TLS in younger children can be misleading and should therefore be considered with caution in selecting patients for surgical procedures on the temporal lobe. PMID- 7489694 TI - Absence epilepsies. AB - Individuals fulfilling diagnostic criteria for childhood absence epilepsy (CAE) and juvenile absence epilepsy (JAE) were selected from a large group of patients who were born between 1945 and 1973 and had presented with absence seizures (AS). Updated data allowed an analysis of 52 patients with CAE and of 62 patients with JAE age > or = 20 years. In CAE, complete control was achieved in 90% of patients (95%, AS only; 77%, AS + generalized tonic-clonic seizures, GTCS). Only 16% of patients with an onset < 9 years had developed GTCS. In JAE, complete control was achieved in 37% of patients (47%, AS; 37%, GTCS). These figures support the validity of the International Classification of Epilepsy (ICE). Stricter diagnostic criteria are discussed. PMID- 7489697 TI - Gabapentin toxicity in children manifesting as behavioral changes. AB - Pediatric experience with gabapentin (GBP), a new antiepileptic drug (AED), is limited. We described 3 learning disabled children, 1 aged 7 and 2 aged 10 years, with intractable partial seizures who developed severe behavioral problems while receiving modest doses of GBP. The children became hyperactive and had explosive outburst consisting of aggressive and oppositional behavior. The behavioral problems were sufficiently severe to require discontinuation of GBP despite moderately improved seizure control. PMID- 7489695 TI - Sudden death in epilepsy: a study of incidence in a young cohort with epilepsy and learning difficulty. AB - Sudden death, often seizure related, may occur in patients with epilepsy. Population-based incidence is probably on the order of 1:1,000/year. The incidence is much higher in selected groups, however. We wished to establish the incidence of sudden unexpected death (SUD) in a young cohort with severe epilepsy and learning difficulties. The study cohort included 310 pupils with epilepsy enrolled at a special residential school between April 1970 and April 1993. The follow-up period totaling 4,135 person-years included a period of residence at the school as well as time after leaving. Age and sex standardized overall mortality ratio was 15.9 [95% confidence interval (CI) 10.6-23.0], with 20 of 28 deaths considered epilepsy related. An incidence of sudden death cases of 1:295/year was noted. All 14 sudden deaths occurred when the pupils were not under the close supervision of the school and most were unwitnessed, which has implications for prevention. PMID- 7489696 TI - Comparing the cognitive effects of phenytoin and carbamazepine in long-term monotherapy: a two-year follow-up. AB - We compared the cognitive effects of randomly prescribed phenytoin (PHT) and carbamazepine (CBZ) therapy on newly diagnosed patients with epilepsy in a 2-year parallel group follow-up study. Fifteen patients were receiving PHT and 16 were receiving CBZ. Neuropsychological assessments were conducted before the treatment and after 6 and 24 months of steady-state drug therapy. Differential effects of PHT and CBZ during follow-up were observed in 3 of 32 measurements. PHT appeared to have negative effects on visually guided motor speed of both hands. In addition, the performance of the PHT group as compared with the CBZ group developed less positively in one visual memory task. The development of mood as measured by Profile of Mood States (POMS), was quite similar in both drug groups; Tension, Depression, and Bewilderment decreased and Vigor increased during the follow-up. The results suggest that the long-term effects of PHT as compared with those of CBZ on cognition are few and restricted mainly to some visually guided motor functions. The effects of PHT on cerebellar function as a possible mechanism for these changes is discussed. PMID- 7489698 TI - 4p(-) syndrome: a chromosomal disorder associated with a particular EEG pattern. AB - We report an electroclinical and cytogenetic study of 4 patients with Wolf Hirschhorn syndrome (WHS). In all cases, we observed a stereotyped EEG and clinical picture characterized by generalized or unilateral myoclonic seizures followed later by brief atypical absences. Electrographically, these were accompanied by a sequence of centroparietal or parietotemporal sharp waves; high voltage wave with a superimposed spike becoming unusual spike-wave complexes, often elicited by eye closure; burst of diffuse spikes and waves; and frequent jerks. This electroclinical pattern is very similar to the one described in Angelman syndrome (AS) in which a defect in GABAA receptor function has been suggested. Moreover, the genes encoding the GABAA receptor subunit have been mapped to the p12-p13 bands of chromosome 4. Even though the deletion in these cases does not encompass the 4p12-p13 region, we suggest that the electroclinical picture common to WHS and AS might represent a characteristic type of epilepsy linked to a common genetic abnormality. PMID- 7489699 TI - Hemimegalencephaly: localized proton magnetic resonance spectroscopy in vivo. AB - Two children with hemimegalencephaly were examined by magnetic resonance imaging (MRI) and localized proton MR spectroscopy (MRS). In both cases, structural changes in the enlarged hemisphere included pachy- or polymicrogyria and gliosis of white matter. Associated metabolic disturbances included a dramatic reduction of glutamate and N-acetylaspartate (NAA) in white matter. Less severe or no alterations were noted in cortical gray matter, basal ganglia, and cerebellum. The older child (13 years) showed increased myoinositol in both gray and white matter as well as markedly increased choline-containing compounds in gray matter. Both children also had mildly decreased NAA levels in the white matter of the contralateral hemisphere. The spectroscopic findings indicate loss of vital neuroaxonal tissue and glial cell proliferation. Metabolic disturbances were more pronounced in the older child. The normal-appearing hemisphere was mildly affected in both cases. PMID- 7489700 TI - [11C]flumazenil positron emission tomography visualizes frontal epileptogenic regions. AB - Presently available noninvasive methods correctly localize epileptogenic regions in only approximately 50% of patients with frontal lobe epilepsy (FLE). Earlier studies have shown that temporal lobe epileptogenic regions may be identified readily by positron emission tomography (PET) measurements of regional benzodiazepine (BZD) receptor binding. We tested the specific applicability of this method in patients with FLE. Six patients with frontal partial seizures and 7 healthy men were investigated with PET and the BZD receptor ligand [11C]flumazenil. All patients had magnetic resonance (MR) brain scans. The independent assessment of seizure-onset region was based on seizure semiology, intra- and extracranial EEG and, in 4 cases, also on [18F]fluorodeoxyglucose (FDG)-PET. The epileptic focus/seizure-generating region was correctly identified by [11C]flumazenil PET in all patients. This region was characterized by a significant reduction in BZD receptor density. The area with reduced BZD receptor density was better delimited than the corresponding hypometabolic region, which was observed in 50% of patients investigated with [18F]FDG-PET. MRI was normal in 5 patients. Visualization of BZD receptors with [11C]flumazenil PET appears to be a promising approach for noninvasive identification of frontal lobe epileptogenic regions. PMID- 7489701 TI - Case of simple partial status epilepticus in occipital lobe epilepsy misdiagnosed as migraine: clinical, electrophysiological, and magnetic resonance imaging characteristics. AB - An 31-year-old man had a unique form of occipital lobe epilepsy. Since age 13 years, he has had episodes of simple partial status epilepticus (SE) occurring twice a month. These typically consisted of elementary visual hallucinations of flashing lights obscuring his left visual field for a period of 2 days, associated with a severe frontal headache initially diagnosed as migraine. These episodes of simple partial SE then evolved to a complex partial seizure (CPS) or secondarily generalized seizure. There were unique EEG features, including: (a) the perception of a flash of light in the left visual field with a single sharp/slow wave discharge over the right occipital lobe, (b) right occipital lobe epileptiform activity during the prolonged aura, and (c) an abnormal response to photic stimulation, with occipital lobe discharges during low rates of stimulation (3-5 Hz), time-locked to the stimulus. High-resolution magnetic resonance imaging (MRI) with quantitative morphometry demonstrated that the right hemisphere and right caudate nucleus were smaller than those on the left. An abnormal gyral pattern was also noted over the right parietal region. Occasionally, distinguishing occipital lobe epilepsy from migraine may be difficult. PMID- 7489702 TI - Cerebral neurocytoma: an unusual cause of refractory epilepsy. Case report and review of the literature. AB - We report a patient with medically refractory complex partial seizures (CPS) caused by a cerebral neurocytoma located near the amygdala. Neurocytoma represents an important addition to the differential diagnosis and, in particular, must be differentiated from oligodendroglioma and dysembryoplastic neuroepithelial tumor. Accurate pathological differential has therapeutic and prognostic implications. PMID- 7489703 TI - Propofol in prehospital treatment of convulsive status epilepticus. AB - We studied the safety and efficacy of intravenous propofol in the out-of-hospital treatment of convulsive status epilepticus (SE) in 8 patients (age 29-70 years), 4 of them with posttraumatic epilepsy. Four patients had no history of seizures. Convulsions ceased promptly after patients received a bolus of 100-200 mg propofol administered before hospital admission by staff of a mobile intensive care unit (ICU). The median duration of coma was 3 h 15 min (range 2-41 h), and the median duration of hospital treatment was 3 1/2 days (range 12 h to 23 days). Only 1 patient was admitted to the hospital's ICU. No adverse effects was observed except for a transient decrease in systolic blood pressure (SBP). Propofol may be a useful drug for the prehospital treatment of recurrent seizures not responding to intravenous diazepam (DZP). PMID- 7489704 TI - Two heads are better than one: crystal structure of the insect derived double domain Kazal inhibitor rhodniin in complex with thrombin. AB - Rhodniin is a highly specific inhibitor of thrombin isolated from the assassin bug Rhodnius prolixus. The 2.6 Angstrum crystal structure of the non-covalent complex between recombinant rhodniin and bovine alpha-thrombin reveals that the two Kazal-type domains of rhodniin bind to different sites of thrombin. The amino terminal domain binds in a substrate-like manner to the narrow active-site cleft of thrombin; the imidazole group of the P1 His residue extends into the S1 pocket to form favourable hydrogen/ionic bonds with Asp189 at its bottom, and additionally with Glu192 at its entrance. The carboxy-terminal domain, whose distorted reactive-site loop cannot adopt the canonical conformation, docks to the fibrinogen recognition exosite via extensive electrostatic interactions. The rather acidic polypeptide linking the two domains is displaced from the thrombin surface, with none of its residues involved in direct salt bridges with thrombin. The tight (Ki = 2 x 10(-13) M) binding of rhodniin to thrombin is the result of the sum of steric and charge complementarity of the amino-terminal domain towards the active-site cleft, and of the electrostatic interactions between the carboxy terminal domain and the exosite. PMID- 7489705 TI - Expression of band 3 anion exchanger induces chloride current and taurine transport: structure-function analysis. AB - Most, but not all, cell types release intracellular organic solutes (e.g. taurine) in response to cell swelling to achieve cell volume regulation. Although this efflux is blocked by classical inhibitors of the electroneutral anion exchanger band 3 (AE1), it is thought to involve an anion channel. The role of band 3 in volume-dependent taurine transport was determined by expressing, in Xenopus oocytes, band 3 from erythrocytes which do (trout) or do not (mouse) release taurine when swollen. AE1 of both species elicited anion exchange activity, but only trout band 3 showed chloride channel activity and taurine transport. Chimeras constructed from trout and mouse band 3 allowed the identification of some protein domains critically associated with channel activity and taurine transport. The data provide evidence that swelling-induced taurine movements occur via an anion channel which is dependent on, or controlled by, band 3. They suggest the involvement of proteins of the band 3 (AE) family in cell volume regulation. PMID- 7489706 TI - A prokaryotic potassium ion channel with two predicted transmembrane segments from Streptomyces lividans. AB - We report the identification, functional expression, purification, reconstitution and electrophysiological characterization of an up to now unique prokaryotic potassium ion channel (KcsA). It has a rectifying current-voltage relationship and displays subconductance states, the largest of which amounts to A approximately equal to 90 pS. The channel is blocked by Cs- ions and gating requires the presence of Mg2+ ions. The kcsA gene has been identified in the gram positive soil bacterium Streptomyces lividans. It encodes a predicted 17.6 kDa protein with two potential membrane-spanning helices linked by a central domain which shares a high degree of similarity with the H5 segment conserved among eukaryotic ion channels. Multiple alignments of deduced amino acids suggest that the novel channel has the closest kinship to the S5, H5 and S6 regions of voltage gated K+ channel families, mainly to the subfamily represented by the Shaker protein from Drosophila melanogaster. Moreover, KcsA is most distantly related to eukaryotic inwardly rectifying channels with two putative predicted transmembrane segments. PMID- 7489707 TI - Microfilament reorganization during apoptosis: the role of Gas2, a possible substrate for ICE-like proteases. AB - Gas2, a component of the microfilament system, belongs to the class of gas genes whose expression is induced at growth arrest. After serum or growth factor addition to quiescent NIH 3T3 cells, Gas2 is hyperphosphorylated and relocalized at the membrane ruffles. By overexpressing gas2wt and a series of deletion mutants of the C-terminal region, we have analysed its role in the organization of the actin cytoskeleton in different cell lines. Overexpression of Gas2 deleted at its C-terminal region (delta 276-314 and delta 236-314), but not its wild-type form, induces dramatic changes in the actin cytoskeleton and cell morphology. These effects are not due to interference of the deleted forms with the endogenous Gas2wt function but could be ascribed to a gain of function. We demonstrate that during apoptosis the C-terminal domain of Gas2 is removed by proteolytic cleavage, resulting in a protein that is similar in size to the described delta 276-314. Moreover, by using in vitro mutagenesis, we also demonstrate that the proteolytic processing of Gas2 during apoptosis is dependent on an aspartic acid residue at position 279. The evidence accumulated here could thus represent a first example of a mechanism linking apoptosis with the co ordinated microfilament-dependent cell shape changes, as possibly mediated by an interleukin-1 beta-converting enzyme (ICE)-like dependent proteolytic cleavage of the Gas2 protein. PMID- 7489708 TI - Cell-free reconstitution of Fas-, UV radiation- and ceramide-induced apoptosis. AB - Cell-free systems are valuable tools for the dissection of complex cellular processes. Here we show that cytoplasmic extracts from cells exposed to anti-Fas antibody or UV radiation contain an activity capable of reproducing morphological changes typical of apoptosis in nuclei added to these extracts, as well as internucleosomal cleavage of DNA and proteolysis of a protein known to be cleaved during the apoptosis of intact cells. Extracts from control cell populations were inactive in this respect. These effects were partly blocked by the addition of purified Bcl-2 protein or a competitive inhibitor peptide of interleukin-1 beta converting enzyme to the extracts. Furthermore, apoptotic activity was induced in cytoplasmic extracts from untreated cells by the addition of ceramide, a lipid second messenger implicated recently in apoptosis signaling. These extracts should prove highly useful in the dissection of molecular events that occur during apoptosis. PMID- 7489709 TI - Apoptosis by a cytosolic extract from Fas-activated cells. AB - Fas is a type I membrane protein and its activation by binding of the Fas ligand or an agonistic anti-Fas antibody induces apoptosis in Fas-bearing cells. In this report we prepared lysates from cells treated with anti-Fas antibody. The lysates induced apoptotic morphological changes in nuclei from normal mouse liver, accompanied by DNA degradation. The apoptosis-inducing activity was quickly generated in cells by anti-Fas antibody and was found in the soluble cytosolic fraction. Induction of the activity in cells was inhibited by a tetrapeptide, acetyl-Tyr-Val-Ala-Asp-chloromethylketone, a specific inhibitor of interleukin-1 beta converting enzyme. Addition of COS cell lysates containing Bcl-2 to the assay significantly inhibited the apoptotic process, indicating that the in vitro process reflected apoptosis that occurs in intact cells. PMID- 7489710 TI - Targeted disruption of the housekeeping gene encoding glucose 6-phosphate dehydrogenase (G6PD): G6PD is dispensable for pentose synthesis but essential for defense against oxidative stress. AB - Glucose 6-phosphate dehydrogenase (G6PD) is a housekeeping enzyme encoded in mammals by an X-linked gene. It has important functions in intermediary metabolism because it catalyzes the first step in the pentose phosphate pathway and provides reductive potential in the form of NADPH. In human populations, many mutant G6PD alleles (some present at polymorphic frequencies) cause a partial loss of G6PD activity and a variety of hemolytic anemias, which vary from mild to severe. All these mutants have some residual enzyme activity, and no large deletions in the G6PD gene have ever been found. To test which, if any, function of G6PD is essential, we have disrupted the G6PD gene in male mouse embryonic stem cells by targeted homologous recombination. We have isolated numerous clones, shown to be recombinant by Southern blot analysis, in which G6PD activity is undetectable. We have extensively characterized individual clones and found that they are extremely sensitive to H2O2 and to the sulfydryl group oxidizing agent, diamide. Their markedly impaired cloning efficiency is restored by reducing the oxygen tension. We conclude that G6PD activity is dispensable for pentose synthesis, but is essential to protect cells against even mild oxidative stress. PMID- 7489711 TI - Expression of a dominant negative mutant of epidermal growth factor receptor in the epidermis of transgenic mice elicits striking alterations in hair follicle development and skin structure. AB - Epidermal growth factor receptor (EGFR) is a key regulator of keratinocyte biology. However, the physiological role of EGFR in vivo has not been well established. To analyze the role of EGFR in skin, we have generated transgenic mice expressing an EGFR dominant negative mutant in the basal layer of epidermis and outer root sheath of hair follicles. Mice expressing the mutant receptor display short and waved pelage hair and curly whiskers during the first weeks of age, but subsequently pelage and vibrissa hairs become progressively sparser and atrophic. Eventually, most mice present severe alopecia. Histological examination of the skin of transgenic mice shows striking alterations in the development of hair follicles, which fail to enter into catagen stage. These alterations eventually lead to necrosis and disappearance of the follicles, accompanied by strong infiltration of the skin with inflammatory elements. The interfollicular epidermis of these mice shows marked hyperplasia, expression of hyperproliferation-associated keratin K6 and increased 5-bromo-2-deoxyuridine incorporation. EGFR function was inhibited in transgenic skin keratinocytes, since in vivo and in vitro autophosphorylation of EGFR was almost completely abolished on EGF stimulation. These results implicate EGFR in the control of hair cycle progression, and provide new information about its role in epidermal growth and differentiation. PMID- 7489712 TI - A synthetic, chemically modified ribozyme eliminates amelogenin, the major translation product in developing mouse enamel in vivo. AB - Ribozymes are small RNA structures capable of cleaving RNA target molecules in a catalytic fashion. Designed ribozymes can be targeted to specific mRNAs, blocking their expression without affecting normal functions of other genes. Because of their specific and catalytic mode of action ribozymes are ideal agents for therapeutic interventions against malfunctioning or foreign gene products. Here we report successful experiments to 'knock out' a major translation product in vivo using synthesized, chemically modified ribozymes. The ribozymes, designed to cleave amelogenin mRNA, were injected close to developing mandibular molar teeth in newborn mice, resulting in a prolonged and specific arrest of amelogenin synthesis not caused by general toxicity. No carriers were required to assist cellular uptake. Amelogenins are highly conserved tissue-specific proteins that play a central role in mammalian enamel biomineralization. Ultrastructural analyses of in vivo ribozyme-treated teeth demonstrated their failure to develop normally mineralized enamel. These results demonstrate that synthesized ribozymes can be highly effective in achieving both timed and localized 'knock-out' of important gene products in vivo, and suggest new possibilities for suppression of gene expression for research and therapeutic purposes. PMID- 7489714 TI - sepB: an Aspergillus nidulans gene involved in chromosome segregation and the initiation of cytokinesis. AB - In Aspergillus nidulans conidia, cytokinesis (septation) is delayed until three rounds of nuclear division have been completed. This has permitted the identification of essential genes that are involved in the coordination of cytokinesis with nuclear division. Conditional mutations in the sepB gene block septation but allow germinating spores to complete the first three rounds of nuclear division at restrictive temperature. sepB3 mutants demonstrate transient delays in M-phase, accumulate aneuploid nuclei and show defects in chromosome segregation. Molecular analysis of the sepB gene reveals that it is essential and possesses limited similarity to the CTF4 gene of Saccharomyces cerevisiae. Using temperature-shift analysis we show that sepB is required after the first nuclear division but before the onset of cytokinesis. A failure to execute the sepB function results in a block to nuclear division and leads to cell death at a time when wild-type cells would be undergoing cytokinesis. Finally, we demonstrate that sepB is also required for the uninucleate cell divisions of developing conidiophores. Our results suggest that sepB3 mutants accumulate specific nuclear defects that do not arrest mitosis, but block the initiation of septum formation. Thus, proper chromosome segregation and a functional sepB gene are required to initiate cytokinesis. PMID- 7489713 TI - The role of gsc and BMP-4 in dorsal-ventral patterning of the marginal zone in Xenopus: a loss-of-function study using antisense RNA. AB - The dorsal-specific homeobox gene goosecoid (gsc) and the bone morphogenetic protein 4 gene (BMP-4) are expressed in complementary regions of the Xenopus gastrula. Injection of gsc mRNA dorsalizes ventral mesodermal tissue and can induce axis formation in normal and UV-ventralized embryos. On the other hand, BMP-4 mRNA injection, which has a strong ventralizing effect on whole embryos, has been implicated in ventralization by UV, and can rescue tail structures in embryos dorsalized by LiCl. The above-mentioned putative roles for BMP-4 and gsc are based on gain-of-function experiments. In order to determine the in vivo role of these two genes in the patterning of the Xenopus mesoderm during gastrulation, partial loss-of-function experiments were performed using antisense RNA injections. Using marker genes that are expressed early in gastrulation, we show that antisense gsc RNA has a ventralizing effect on embryos, whereas antisense BMP-4 RNA dorsalizes mesodermal tissue. These loss-of-function studies also show a requirement for gsc and BMP-4 in the dorsalization induced by LiCl and in the ventralization generated by UV irradiation, respectively. Thus, both gain- and loss-of-function results for gsc and BMP-4 support the view that these two genes are necessary components of the dorsal and ventral patterning pathways in Xenopus embryos. PMID- 7489716 TI - The mating-type locus B alpha 1 of Schizophyllum commune contains a pheromone receptor gene and putative pheromone genes. AB - Analysis of the multispecific B alpha mating-type locus of Schizophyllum commune provided evidence that pheromones and pheromone receptors govern recognition of self versus non-self and sexual development in this homobasidiomycetous fungus. Four subclones of an 8.2 kb genomic fragment carrying B alpha 1 specificity induced B-regulated sexual morphogenesis when introduced into a strain with one of the eight compatible B alpha specificities that are known to exist in nature. One of these clones, which activated all other B alpha specificities, contains a gene termed bar1. The predicted protein product of bar1, as well as that of bar2, a homologous gene isolated from a B alpha 2 strain, has significant homology to known fungal pheromone receptor proteins in the rhodopsin-like superfamily of G protein-linked receptors. The other three active B alpha 1 clones were subcloned further to identify the minimal active element in each clone. Every active subclone contains a putative pheromone gene ending in a signal for possible isoprenylation. A message of approximately 600 bp was observed for one of these genes, bap1(1). This paper presents the first evidence for a system of multiple pheromones and pheromone receptors as a basis for multispecific mating types in a fungus. PMID- 7489715 TI - The GTPase Ypt7p of Saccharomyces cerevisiae is required on both partner vacuoles for the homotypic fusion step of vacuole inheritance. AB - In the budding yeast Saccharomyces cerevisiae, vacuoles are inherited by the projection of vesicles and tubules from the mother-cell vacuole into the growing daughter cell during the S phase. These vesicles then fuse and form the daughter cell organelle. We have described previously in vitro reactions of the formation of vacuole-derived segregation structures and of vacuole-vacuole fusion. Homotypic vacuole fusion requires cytosol, ATP and a physiological temperature, and is sensitive to GTPase inhibitors. These reactions are divisible into early stages which require ATP and cytosol, and late stages which require neither. Here, we report that Ypt7p, a ras-like GTPase implicated previously in endocytosis in yeast, is largely localized to the vacuole and is required on both partners during the in vitro vacuole fusion reaction. The in vitro fusion reaction is inhibited either by Gdi1p, which extracts the GDP-bound form of ras like GTPases from membranes, or by antibodies specific for Ypt7p. The presence of anti-Ypt7p during the early stages of the reaction inhibits the development of cytosol- and ATP-independent intermediates. Although cytosol and ATP are no longer needed for the late stage of vacuole inheritance in vitro, the inhibition of this late stage by anti-Ypt7p or Gdi1p requires the continued presence of ATP and cytosol. Ypt7p is the first GTPase for which a direct role in organelle inheritance has been established. PMID- 7489717 TI - The principal target of rapamycin-induced p70s6k inactivation is a novel phosphorylation site within a conserved hydrophobic domain. AB - The immunosuppressive agent rapamycin induces inactivation of p70s6k with no effect on other mitogen-activated kinases. Here we have employed a combination of techniques, including mass spectrometry, to demonstrate that this effect is associated with selective dephosphorylation of three previously unidentified p70s6k phosphorylation sites: T229, T389 and S404. T229 resides at a conserved position in the catalytic domain, whose phosphorylation is essential for the activation of other mitogen-induced kinases. However, the principal target of rapamycin-induced p70s6k inactivation is T389, which is located in an unusual hydrophobic sequence outside the catalytic domain. Mutation of T389 to alanine ablates kinase activity, whereas mutation to glutamic acid confers constitutive kinase activity and rapamycin resistance. The importance of this site and its surrounding motif to kinase function is emphasized by its presence in a large number of protein kinases of the second messenger family and its conservation in putative p70s6k homologues from as distantly related organisms as yeast and plants. PMID- 7489718 TI - Distinct signal transduction through the tyrosine-containing domains of the granulocyte colony-stimulating factor receptor. AB - The receptor for granulocyte colony-stimulating factor (G-CSFR) is a hemopoietic growth factor receptor, which mediates proliferation and differentiation signals. The cytoplasmic region of G-CSFR carries four tyrosine residues in its C-terminal half. We constructed mutant receptors in which each tyrosine residue of G-CSFR was mutated to phenylalanine. Two mutant receptors (Tyr703 and Tyr728) neither transduced the growth-inhibitory signal nor induced the neutrophil-specific myeloperoxidase (MPO) gene. The Tyr703 mutant did not induce morphological changes in cells, whereas transformants expressing the Tyr728 mutant adhered to plates with a macrophage-like morphology upon G-CSF stimulation. Mutation of the most distal tyrosine residue (Tyr763) abolished the ability of G-CSFR to stimulate the tyrosine phosphorylation of a cellular protein with an M(r) of 54 kDa. These results indicated that the regions around the three tyrosine residues of G-CSFR play essential and distinct roles in signal transduction. PMID- 7489719 TI - Rac GTPase interacts with GAPs and target proteins through multiple effector sites. AB - Rac, a small GTPase in the ras superfamily, regulates at least two biological processes in animal cells: (i) the polymerization of actin and the assembly of integrin complexes to produce lamellipodia and ruffles; and (ii) the activity of an NADPH oxidase in phagocytic cells. NADPH oxidase activation is mediated through a rac effector protein, p67phox, and using chimeras made between rac and the closely related GTPase, rho, we have identified two distinct effector sites in rac, one N-terminal and one C-terminal, both of which are required for activation of p67phox. The same two effector sites are essential for rac-induced actin polymerization in fibroblasts. p65PAK, a ubiquitous serine/threonine kinase, interacts with rac at both the N- and C-terminal effector sites, but the GTPase-activating protein, bcr interacts with rac at a different region. This makes p65PAK, but not bcr, a candidate effector of rac-induced lamellipodium formation. PMID- 7489720 TI - The Drosophila fork head domain protein crocodile is required for the establishment of head structures. AB - The fork head (fkh) domain defines the DNA-binding region of a family of transcription factors which has been implicated in regulating cell fate decisions across species lines. We have cloned and molecularly characterized the crocodile (croc) gene which encodes a new family member from Drosophila. croc is expressed in the head anlagen of the blastoderm embryo under the control of the anterior, the dorsoventral and the terminal maternal organizer systems. The croc mutant phenotype indicates that the croc wild-type gene is required to function as an early patterning gene in the anterior-most blastoderm head segment anlage and for the establishment of a specific head skeletal structure that derives from the non adjacent intercalary segment at a later stage of embryogenesis. As an early patterning gene, croc exerts unusual properties which do not allow it to be grouped among the established segmentation genes. A single-site mutation within the croc fkh domain, which causes a replacement of the first out of four conserved amino acid residues thought to be involved in the coordinate binding of Mg2+, abolishes the DNA binding of the protein in vitro. In view of the resulting lack-of-function mutant phenotype, it appears likely that metal binding by the affected region of the fkh domain is crucial for proper folding of the DNA binding structure. PMID- 7489722 TI - Extending dimerization interfaces: the bZIP basic region can form a coiled coil. AB - We appended a rationally designed acidic amphipathic protein sequence to the N terminus of a leucine zipper. Circular dichroism data indicate that this engineered polypeptide sequence can 'zipper' up the basic region of a bZIP monomer into a heterodimeric coiled coil. This propagation of the leucine zipper dimerization interface into the basic region can proceed for up to four heptads and stabilizes the heterodimer complex 2.5 kcal/mol or > 100-fold. The acidic nature of the extension is the most critical component of the design, suggesting that the extension is acting as a DNA mimetic. The dimerization prevents the basic region in this heterodimeric coiled coil structure from binding to DNA. Gel shift, fluorescence and transient transfection assays indicate that the acidic extension appended to a leucine zipper can inactivate the DNA-binding and transactivation properties of the bZIP protein C/EBP. The three bZIP basic regions examined in this study dimerize with similar stability with the acidic extension, suggesting that this N-terminal extension can be used to develop dominant-negatives to other bZIP transcription factors. PMID- 7489721 TI - The reduced expression of endogenous duplications (REED) in the maize R gene family is mediated by DNA methylation. AB - The duplicated R and Sn genes regulate the maize anthocyanin biosynthetic pathway and encode tissue-specific products that are homologous to helix-loop-helix transcriptional activators. As a consequence of their coupling in the genome, Sn is partially silenced. Genomic restriction analysis failed to reveal gross structural DNA alterations between the strong original phenotype and the weak derivatives. However, the differences in pigmentation were inversely correlated with differences in the methylation of the Sn promoter. Accordingly, treatment with 5-azacytidine (AZA), a demethylating agent, restored a strong pigmentation pattern that was transmitted to the progeny and that was correlated with differential expression of the Sn transcript. Genomic sequencing confirmed that methylation of the Sn promoter was more apparent in the less pigmented seedlings and was greatly reduced in the AZA revertants. In addition, some methylcytosines were located in non-symmetrical C sequences. These findings provide an insight into Sn and R interaction, a process that we have termed Reduced Expression of Endogenous Duplications (REED). We propose that increasing the copy number of regulatory genes by endogenous duplication leads to such epigenetic mechanisms of silencing. Further understanding of the REED process may have broader implications for gene regulation and may identify new levels of regulation within eukaryotic genomes. PMID- 7489723 TI - Fos is an essential component of the mammalian UV response. AB - Mouse 3T3 fibroblasts lacking c-fos were employed to demonstrate an essential function of the UV-inducible transcription factor AP-1 (Fos/Jun) in the response to the cytotoxic effects of short-wavelength ultraviolet (UVC) radiation. Clonogenic survival and proliferation of cells lacking c-fos were drastically reduced following UV irradiation. This UV hypersensitivity manifests itself primarily in increased cell death, partly by apoptosis, and prolonged recovery time from UV-induced cell cycle arrest. Co-culture with wild-type cells did not ameliorate the hypersensitivity of mutant cells. Transcriptional induction of the c-Fos target genes collagenase I, stromelysin-1 and stromelysin-2 by UV is almost absent in cells lacking c-fos which correlates with a reduced UV induction of AP 1 DNA-binding and transactivation activity. The repair of UV-induced DNA lesions was not affected, as shown by unscheduled DNA synthesis and host cell reactivation assays. These data demonstrate that c-Fos is involved in a novel protective function other than DNA repair against the harmful consequences of UVC. PMID- 7489724 TI - Requirements for iron-regulated degradation of the RNA binding protein, iron regulatory protein 2. AB - Iron regulatory proteins (IRPs) regulate the expression of genes involved in iron metabolism whose transcripts contain RNA stem-loop motifs known as iron responsive elements (IREs). When iron concentrations are low, IRPs bind to IREs in the 5' untranslated region (UTR) of transcripts where they repress translation, or the 3' UTR of transcripts where they inhibit degradation. The RNA binding activities of the homologous proteins IRP1 and IRP2 are both regulated post-translationally. The binding activity of IRP2 is regulated by the degradation of the protein when cells are iron-replete. Here, we demonstrate that a 73 amino acid sequence that corresponds to a unique exon in IRP2 contains a sequence required for rapid degradation in iron-replete cells. The deletion of this sequence eliminates the rapid turnover of IRP2, whereas the transfer of this sequence to the corresponding position in the homologous protein IRP1 confers the capacity for iron-dependent degradation upon IRP1. Site-directed mutagenesis has demonstrated that specific cysteines within the IRP2 exon are required for iron dependent degradation. The degradation of IRP2 appears to be mediated by the proteasome in iron-replete cells. When degradation is prevented, the RNA binding activity of IRP2 is not regulated by iron concentration. Thus, degradation is required for the regulation of the RNA binding activity of IRP2. PMID- 7489725 TI - The fragile X mental retardation syndrome protein interacts with novel homologs FXR1 and FXR2. AB - Fragile X Mental Retardation Syndrome is the most common form of hereditary mental retardation, and is caused by defects in the FMR1 gene. FMR1 is an RNA binding protein and the syndrome results from lack of expression of FMR1 or expression of a mutant protein that is impaired in RNA binding. The specific function of FMR1 is not known. As a step towards understanding the function of FMR1 we searched for proteins that interact with it in vivo. We have cloned and sequenced a protein that interacts tightly with FMR1 in vivo and in vitro. This novel protein, FXR2, is very similar to FMR1 (60% identity). FXR2 encodes a 74 kDa protein which, like FMR1, contains two KH domains, has the capacity to bind RNA and is localized to the cytoplasm. The FXR2 gene is located on human chromosome 17 at 17p13.1. In addition, FMR1 and FXR2 interact tightly with the recently described autosomal homolog FXR1. Each of these three proteins is capable of forming heteromers with the others, and each can also form homomers. FXR1 and FXR2 are thus likely to play important roles in the function of FMR1 and in the pathogenesis of the Fragile X Mental Retardation Syndrome. PMID- 7489726 TI - Mutants in a yeast Ran binding protein are defective in nuclear transport. AB - Ran, a Ras-like GTPase, has been implicated in controlling the movement of proteins and RNAs in and out of the nucleus. We have constructed strains of Saccharomyces cerevisiae which produce fusion proteins containing glutathione-S transferase (GST) fused to Gsp1p, which encodes the essential yeast Ran homolog, and a mutant form of Gsp1p that mimics the GTP-bound state. A major protein with the apparent size of 34 kDa co-purifies with the GTP-bound form of Gsp1p. This protein was identified as Yrb1p (Yeast Ran Binding Protein) and stimulates GTP hydrolysis by Gsp1p in the presence of Rna1p, the Gsp1 GTPase activating protein. Yrb1p is located in the cytoplasm with some concentration at the nuclear periphery. Temperature-sensitive yrb1 mutants are defective in nuclear protein import and RNA export. A mutation in the highly conserved Ran binding region of Yrb1p reduces its ability to interact with Gsp1p. These data indicate that Yrb1p functions with Gsp1p and suggest that together they can control transport of macromolecules across the nuclear envelope. PMID- 7489727 TI - The N-terminal domain of human DNA ligase I contains the nuclear localization signal and directs the enzyme to sites of DNA replication. AB - DNA replication in mammalian cells occurs in discrete nuclear foci called 'replication factories'. Here we show that DNA ligase I, the main DNA ligase activity in proliferating cells, associates with the factories during S phase but displays a diffuse nucleoplasmic distribution in non-S phase nuclei. Immunolocalization analysis of both chloramphenicol acetyltransferase (CAT)-DNA ligase I fusion proteins and epitope tagged DNA ligase I mutants allowed the identification of a 13 amino acid functional nuclear localization signal (NLS) located in the N-terminal regulatory domain of the protein. Furthermore, the NLS is immediately preceded by a 115 amino acid region required for the association of the enzyme with the replication factories. We propose that in vivo the activity of DNA ligase I could be modulated through the control of its sub nuclear compartmentalization. PMID- 7489728 TI - dpa, a member of the MCM family, is required for mitotic DNA replication but not endoreplication in Drosophila. AB - We have isolated the Drosophila disc proliferation abnormal (dpa) gene, a member of the MCM family of DNA replication factors. Members of this family of proteins are required for DNA replication in yeast. A dpa null mutant dies during pupal stages because imaginal tissues necessary for the formation of the adult fly fail to proliferate normally. Beginning in late embryogenesis BrdU labeling reveals DNA replication defects in mitotically proliferating cells. In contrast, dpa is dispensable for endoreplication, a specialized cell cycle consisting of consecutive rounds of S phases without intervening mitosis. Our studies suggest an essential role for dpa in mitotic DNA replication but not in endoreplication. Thus, dpa is not a general replication factor but may play a specialized regulatory role in DNA replication. PMID- 7489730 TI - IS1-mediated intramolecular rearrangements: formation of excised transposon circles and replicative deletions. AB - A system is described which permits visualization and analysis of a number of molecular species associated with transposition activity of the bacterial insertion sequence, IS1, in vivo. The technique involves induction of an IS1 transposase gene carried by a plasmid which also includes an IS1-based transposable element. It is, in principle, applicable to the identification of transposition intermediates as well as unstable transposition products and those which are not detectable by genetic means. Thirteen novel molecular species were detected after 4 h of induction. Five major species were characterized, based on their behaviour as a function of time, on their hybridization patterns and on the nucleotide sequences of the transposon-backbone junctions. All result from intramolecular IS1 transposition events. The two reciprocal partner products of IS1-mediated deletions, the intramolecular equivalent of co-integrates generated by intermolecular transposition, have been identified. Both carry a single copy of the transposable element and present complementary distributions of deletion endpoints. These results establish, by direct physical means, that adjacent IS1 mediated deletions are accompanied by duplication of the element. A second type of molecule identified was an excised circular copy of the transposon, raising the possibility that IS1 is capable of following an intermolecular transposition pathway, via excised transposon circles, leading to direct insertion. PMID- 7489731 TI - Utilization of ribonucleotides and RNA primers by Tetrahymena telomerase. AB - Telomerase is a ribonucleoprotein (RNP) DNA polymerase involved in telomere synthesis. A short sequence within the telomerase RNA component provides a template for de novo addition of the G-rich strand of a telomeric simple sequence repeat onto chromosome termini. In vitro, telomerase can elongate single-stranded DNA primers processively: one primer can be extended by multiple rounds of template copying before product dissociation. Telomerase will incorporate dNTPs or ddNTPs and will elongate any G-rich, single-stranded primer DNA. In this report, we show that Tetrahymena telomerase was able to incorporate a ribonucleotide, rGTP, into product polynucleotide. Synthesis of the product [d(TT)r(GGGG)]n was processive, suggesting that the chimeric product remained associated with the enzyme both at the active site and at a second, previously characterized, template-independent product binding site. As predicted by this finding, RNA-containing oligonucleotides served as primers for elongation. More than 3 nt of RNA at a primer 3' end decreased the quantity of product synthesis but increased the affinity of the primer for telomerase. Thus, RNA-containing primers were effective as competitive inhibitors of DNA primer elongation by telomerase. These results support the possible evolutionary origin of telomerase as an RNA-dependent RNA polymerase. PMID- 7489729 TI - Preferential binding of human topoisomerase I to superhelical DNA. AB - Eukaryotic type I DNA topoisomerase provides swivels for removing torsional strain from the DNA helix during transcription and DNA replication. Previously it has been shown that the enzyme is associated with actively transcribed genes and replicating DNA. Using an inactive mutant form of the protein containing phenylalanine instead of tyrosine at position 723, we have investigated the binding properties of the protein as a function of substrate topology. A series of filter binding assays indicated that the protein strongly prefers to bind superhelical over completely relaxed SV40 DNA. The ability of a supercoiled DNA to compete against a relaxed DNA for binding increases as the number of superhelical turns in the DNA increases. Since positively supercoiled DNA is bound with the same preference as negatively supercoiled DNA, we hypothesize that topoisomerase I binds preferentially at the nodes created by the crossing of two duplex helices. The preference for binding superhelical DNA is also exhibited by the conserved core domain (amino acids 175-659) which is missing the active site region located near the C-terminus. These results suggest that this core domain may target the enzyme in vivo to regions of torsionally strained superhelical DNA. PMID- 7489732 TI - Mechanisms underlying telomere repeat turnover, revealed by hypervariable variant repeat distribution patterns in the human Xp/Yp telomere. AB - Sequences immediately adjacent to the human Xp/Yp telomere exhibit a high frequency of base substitutional polymorphisms, together with almost complete linkage disequilibrium, to create only a few diverged haplotypes. This sequence divergence has been used to develop a PCR-based system for mapping the distribution of the telomere (TTAGGG) and variant repeats (TGAGGG and TCAGGG) at the proximal end of the telomere repeat array. The distribution of these repeats is extremely variable. Almost all Xp/Yp telomeres are different, indicating a high mutation rate. Some telomere maps associated with the same flanking haplotype show similarities, identifying subsets of telomeres that share a recent common ancestry. Mechanisms underlying the rapid turnover of repeats at the proximal end of the Xp/Yp telomere include intra-allelic processes, such as slippage during replication. Inter-allelic exchanges may occur occasionally, but telomerase activity probably plays only a minor role in the germline turnover of proximally located telomere and variant repeats. PMID- 7489733 TI - Generation of a human X-derived minichromosome using telomere-associated chromosome fragmentation. AB - A linear mammalian artificial chromosome vector will require at least three functional elements: a centromere, two telomeres and replication origins. One route to generate such a vector is by the fragmentation of an existing chromosome. We have previously described the use of cloned telomeric DNA to generate and stably rescue truncated derivatives of a human X chromosome in a somatic cell hybrid. Further rounds of telomere-associated chromosome fragmentation have now been used to engineer a human X-derived minichromosome. This minichromosome is estimated to be < 10 Mb in size. In situ hybridization and molecular analysis reveal that the minichromosome has a linear structure, with two introduced telomere constructs flanking a 2.5 Mb alpha-satellite array. The highly truncated chromosome also retains some chromosome-specific DNA, originating from Xp11.21. There is no significant change in the mitotic stability of the minichromosome as compared with the X chromosome from which it was derived. PMID- 7489734 TI - Differences in the interleukin-2 (IL-2) receptor system in human and mouse: alpha chain is required for formation of the functional mouse IL-2 receptor. AB - Reconstitution with mouse interleukin-2 (IL-2) receptor subunits demonstrated that the mouse IL-2 receptor complex was different from the human complex in the alpha chain requirement for the functional mouse receptor complex. The heterotrimeric complex of the mouse exogenous alpha and beta chains and the endogenous gamma chain on mouse lymphoid BW5147 cells showed the ability to bind IL-2 with high affinity, resulting in IL-2-induced tyrosine phosphorylation of a cytosolic tyrosine kinase, JAK3, which is involved in IL-2-dependent signals. Exogenous introduction of the beta chain with the endogenous gamma chain, however, could neither confer appreciable IL-2 binding nor IL-2-induced signal transduction on BW5147 cells, unlike the human beta gamma heterodimer. Mouse spleen CD8+ cells, not having the alpha chain initially, showed IL-2-dependent cell proliferation only when expression of the alpha chain was induced. Collectively, these results illustrate that the functional mouse IL-2 receptor complex necessarily includes the alpha chain, and that the regulation of CD8+ T cell growth during immune reaction depends upon alpha chain expression. PMID- 7489736 TI - Beta 2-microglobulin with an endoplasmic reticulum retention signal increases the surface expression of folded class I major histocompatibility complex molecules. AB - With beta 2-microglobulin- (beta 2m-) cell lines such as R1E/Db, the surface expression of class I major histocompatibility complex molecules is greatly impaired, and class I molecules that are on the surface are generally misfolded. To determine whether beta 2m must be continually present with the class I heavy chain for the class I molecule to reach the surface in a folded conformation, a sequence encoding an endoplasmic reticulum (ER) retention signal (KDEL) was attached onto the 3' end of a beta 2m cDNA. After this chimeric cDNA was transfected into R1E/Db cells, beta 2m-KDEL protein was detectable by an anti beta 2m serum within the cells but not at the cell surface. Interestingly, R1E/Db cells transfected with beta 2m-KDEL were found to express a high level of conformationally correct Db molecules at the cell surface. This observation implies that beta 2m has a critical and temporal role in the de novo folding of the class I heavy chain. We propose that the critical time for beta 2m association is when the class I molecule is docked with the transporter associated with antigen processing (TAP) and first interacts with peptide. PMID- 7489735 TI - Major histocompatibility complex class I-restricted CD8+ T cells and class II restricted CD4+ T cells, respectively, mediate and regulate contact sensitivity to dinitrofluorobenzene. AB - Contact sensitivity (CS) is a form of delayed-type hypersensitivity to haptens applied epicutaneously and is thought to be mediated, like classical delayed-type hypersensitivity responses, by CD4+ T helper-1 cells. The aim of this study was to identify the effector T cells involved in CS. We studied CS to the strongly sensitizing hapten dinitrofluorobenzene (DNFB) in mice rendered deficient by homologous recombination in either major histocompatibility complex (MHC) class I, MHC class II, or both, and which exhibited deficiencies in, respectively, CD8+, CD4+, or both, T cells. MHC class I single-deficient and MHC class I/class II double-deficient mice, both of which have a drastic reduction in the number of CD8+ T cells, were unable to mount a CS response to DNFB. In contrast, both MHC class II-deficient mice and normal mice treated with an anti-CD4 monoclonal antibody (mAb) developed exaggerated and persistent responses relative to heterozygous control littermates. Furthermore, anti-CD8 mAb depletion of class II deficient mice totally abolished their ability to mount an inflammatory response to DNFB. Removal of residual CD4+ T cells in class II-deficient mice by anti-CD4 mAb treatment did not diminish the intensity of CS. These data clearly demonstrate that class I-restricted CD8+ T cells are sufficient for the induction of CS to DNFB, and further support the idea that MHC class II-restricted CD4+ T cells down-regulate this inflammatory response. PMID- 7489737 TI - Modulation of immunity to Borrelia burgdorferi by ultraviolet irradiation: differential effect on Th1 and Th2 immune responses. AB - Ultraviolet B (UVB) radiation suppresses the delayed-type hypersensitivity (DTH) response to alloantigen by a mechanism involving interleukin (IL)-10. It has been hypothesized, based on this result, that UV irradiation shifts the immune response from a Th1 to a Th2 response. We tested this hypothesis using Borrelia burgdorferi (Bb) as an antigen under conditions where both DTH and antibody responses could be assessed. Mice were irradiated with a single dose of UV and then immunized with Bb in complete Freund's adjuvant (CFA). DTH was assessed by footpad challenge. At various time points thereafter, mice were bled, and the serum antibodies to Bb were quantitated. Only IgG1, IgG2a, and IgG2b were produced in response to Bb. The IgG2a and IgG2b antibody responses, as well as the DTH response to Bb, showed UV dose-dependent reductions after UV irradiation. The primary IgG1 response to Bb was very low and was unaffected by UV irradiation; however, the IgG1 secondary response was elevated in UV-irradiated mice. Injection of anti-IL-10 antibody into UV-irradiated mice within 24 h after UV exposure restored the DTH response, as well as the IgG2a and IgG2b antibody responses. In addition, injecting recombinant murine IL-10 mimicked some of the effects of UV radiation. Our results support the hypothesis that in vivo, UV irradiation down-regulates Th1 immune responses, while leaving Th2 responses intact, and suggest that IL-10 is an important mediator of this effect. PMID- 7489738 TI - Localization in situ of the co-stimulatory molecules B7.1, B7.2, CD40 and their ligands in normal human lymphoid tissue. AB - Functional interactions between B and T lymphocytes are known to depend on the expression of co-stimulatory molecules B7.1/CD80, B7.2/CD86 and their counter receptors CD28 and CTLA4, as well as CD40 and its ligand CD40L. To study the role of these molecules in situ, an immunohistochemical analysis was carried out on normal human lymphoid tissue. In the germinal centers (GC), B7.1 and B7.2 were differentially expressed. In the dark zone, centroblasts were predominantly B7.1+, while centrocytes in the light zone were B7-2+, resulting in reversed gradients of both markers in GC. Follicle mantle cells were negative for B7.1 and B7.2. Macrophages and interdigitating dendritic cells (IDC) in T cell zones both expressed B7.1 and B7.2. Moreover, clusters of B7.2+ T cells were demonstrated in interfollicular areas. Intrafollicular CD4+ T cells in GC, predominantly in the apical light zone, expressed CD28 and CTLA4, as did the majority of interfollicular T cells. CTLA4 showed a striking excentric cytoplasmic staining, which was also seen on T cells activated in vitro. CD40 was expressed on all B cells and more strongly on macrophages and IDC. Moreover, small clusters of T cells in a rim outside the GC showed CD40 expression. CD40L was expressed both on intrafollicular CD4+ T cells as well as on T cells in T cell zones. The differential distribution of co-stimulatory molecules in different compartments of normal human lymphoid tissue in situ indicates that these interactions play a distinctive role in different stages of B cell differentiation and in the immune response. PMID- 7489739 TI - The proto-oncogene Vav product is constitutively tyrosine-phosphorylated in normal human immature T cells. AB - The proto-oncogene Vav product is markedly tyrosine-phosphorylated after the recruitment of various receptors of cells of hematopoietic origin, including mature T cells. Recent studies on Vav-deficient mice have clearly implicated the product of the proto-oncogene Vav in intrathymic T cell development. Vav tyrosine phosphorylation is probably crucial to connect early tyrosine kinase(s) to downstream molecular events leading to cell division and maturation that occur in the thymus. We investigated the tyrosine phosphorylation of Vav in human thymocytes. Immunoblotting experiments demonstrate that, as in mature T cells, tyrosine phosphorylation of Vav is induced following thymocyte stimulation through the T cell receptor. The main observation, however, is that an important fraction of cellular Vav is constitutively tyrosine-phosphorylated in freshly isolated cells. This phenomenon takes place apparently both in the CD4+CD8+ and the more mature CD4+CD8- and CD4-CD8+ thymocyte cell subsets. Co immunoprecipitation experiments showed, moreover, that a small amount of Vav is engaged in the multimolecular complex that includes elements of the T cell receptor and the T cell specific ZAP-70 tyrosine kinase. Altogether, these data suggest that a critical pathway for T cell development in the human thymus likely involves the permanent activation of Vav in vivo. PMID- 7489740 TI - Staphylococcal enterotoxin B and tumor-necrosis factor-alpha-induced relapses of experimental allergic encephalomyelitis: protection by transforming growth factor beta and interleukin-10. AB - A study was made of the ability of the superantigen staphylococcal enterotoxin B (SEB) to induce relapses of experimental allergic encephalomyelitis (EAE) in SJL mice that had partially or completely recovered from acute EAE. We find that a single injection of 0.05 mg SEB i.v. induces mild relapses in 50% of such mice. In addition, tumor necrosis factor (TNF)-alpha (0.2 micrograms, i.p.) also induces EAE relapses in 43% of SJL mice when injected 1-2 months after recovery. SEB does not induce a second relapse if reinjected when V beta 17a+T cells are still partially deleted. In these mice, however, TNF-alpha is equally effective in inducing relapses as in mice that did not receive SEB previously. We showed earlier that transforming growth factor (TGF)-beta and TNF-alpha have antagonistic effects on experimental autoimmune diseases; e.g., in spontaneously relapsing EAE, TGF-beta and anti-TNF were protective, while anti-TGF-beta caused disease exacerbation. Interleukin (IL)-10 is also known to counteract certain TNF effects. We now find that both human IL-10 and TGF-beta 2 lower the incidence of EAE relapses when given simultaneously with SEB or TNF-alpha. The protective effect of TGF-beta is significant only against relapses induced by SEB (reduced to 9%), and that of IL-10 only against relapses induced by TNF (reduced to 0%) with the treatment regimens employed. Neutralizing anti-TGF-beta does not increase the incidence of SEB-induced EAE relapses. In contrast, anti-IL-10 increases both the incidence and the severity of such relapses. We conclude that TNF production is probably important in causing EAE relapses, but that other aspects of the SEB-induced reactivation of myelin-specific T cells also contribute. Furthermore, endogenous IL-10 rather than TGF-beta production appears to limit the susceptibility to induction of EAE relapses in this model. PMID- 7489741 TI - Interleukin-7 can induce the activation of Jak 1, Jak 3 and STAT 5 proteins in murine T cells. AB - The activation of Janus protein tyrosine kinases (Jak) and STAT (signal transducer and activator of transcription) proteins has recently been linked to the signal transduction mechanism of several cytokines. IL-7 was observed to induce a rapid and dose-dependent tyrosine phosphorylation of Jak 1 and Jak 3 and concomitantly, the tyrosine phosphorylation and DNA binding activity of multiple STAT proteins. The STAT proteins utilized by IL-7 were identical to those induced by IL-2 and could be identified as various STAT 5 isoforms. Moreover, the induction of both Jak 1 and 3, and STAT 5 activity strongly correlated with the growth-promoting effects of IL-7, suggesting that this signal transduction mechanism may play a key role in IL-7-induced proliferation. PMID- 7489742 TI - Administration of interleukin-5 or -10 activates peritoneal B-1 cells and induces autoimmune hemolytic anemia in anti-erythrocyte autoantibody-transgenic mice. AB - Activation mechanisms of B-1 (Ly-1 B) cells have been suggested to be different from those of conventional B cells. To assess the role of various interleukins (IL) in the activation of B-1 cells, we injected IL-4, IL-5 or IL-10 into nonanemic anti-red blood cells (RBC) autoantibody-transgenic mice, in which conventional B cells are clonally deleted but peritoneal B-1 cells persist without secreting Ig. Intraperitoneal or intramuscular injection of IL-5 or IL 10, but not IL-4, increased the number of antibody-producing peritoneal B-1 cells by four- to five-fold, resulting in increased anti-RBC serum autoantibody and induction of hemolytic anemia. These results suggest that IL-5 or IL-10 may play an important role in the terminal differentiation of B-1 cells into antibody producing cells in vivo. PMID- 7489743 TI - Generalized autoimmune disease in interleukin-2-deficient mice is triggered by an uncontrolled activation and proliferation of CD4+ T cells. AB - Interleukin-2-deficient mice (IL-2-/-) crossed to a BALB/c genetic background develop a lymphoproliferative syndrome with severe hemolytic anemia and die within 5 weeks of age. The presence of autoantibodies of various specificities and inflammatory lesions in several organs are indicative of a generalized auto immune disease. No alterations of the immune system were observed in 6-day-old animals, but 10-day-old mice already showed an increased proliferation and polyclonal activation of lymphocytes. The treatment of IL-2-/- mice with anti gp39(CD40L) antibody prevented the disease and indicated that the appearance of activated CD4- T cells (CD44high, CD69-) represents the first alteration of the immune system in IL-2-/- mice. Collectively, our results suggest that an essential role of IL-2 in vivo, which is not compensated by other cytokines, is the maintenance of self tolerance. PMID- 7489744 TI - Preferential dependence of autoantibody production in murine lupus on CD86 costimulatory molecule. AB - Blockade of the interactions between CD28/CTLA-4 and their ligands, CD80 (B7, B7.1)/CD86 (B70, B7.2), seems an attractive means to induce antigen-specific peripheral tolerance in organ transplantation and autoimmune disease. Recently, diversities between CD80 and CD86 in expression, regulation, and function have been reported in certain cell populations and murine experimental disease models. To investigate the possible differential role of CD80 and CD86 in the development of lupus, we treated lupus-prone NZB/W F1 mice with specific monoclonal antibodies (mAb) against CD80, CD86, or both. The treatment with a combination of anti-CD80 and CD86 mAb before the onset of lupus completely prevented autoantibody production and nephritis, and prolonged survival. Interestingly, we found that anti-CD86 mAb alone, but not anti-CD80 mAb, efficiently inhibited autoantibody production. Subclass study on IgG anti-double-stranded (ds) DNA antibody revealed that the treatment with anti-CD86 mAb almost completely inhibited both IgG1 and IgG2b, but not IgG2a production. The incomplete reduction of IgG2a anti-dsDNA antibody by anti-CD86 mAb was compensated by the addition of anti-CD80 mAb. A significant reduction of mRNA for interleukin (IL)-2, interferon gamma, IL-4 and IL-6 was observed in mice treated with a combination of anti-CD80 and CD86 mAb or anti-CD86 mAb alone. Treatment with both mAb after the onset of lupus resulted in a significantly prolonged survival with reduction of autoantibody production. These results suggest that CD86 plays a more critical role in autoantibody production, and CD86, but not CD80, contributes to Th2 mediated Ig production. However, the blockade of both CD80 and CD86 are required for preventing the development and progression of lupus. PMID- 7489745 TI - Structural and functional analysis of the promoter of the murine V gamma 1.1 T cell receptor gene. AB - The expression of the germ-line gene V gamma 1.1-C gamma 4 of the T cell receptor (TcR) gamma chain depends on interleukin (IL)-3 induction in hematopoietic cells, while in T cells, the rearranged gene is expressed constitutively. To understand the mechanism that controls TcR gamma gene expression, we cloned and characterized the structure and function of the V gamma 1.1-C gamma 4 TcR promoter. IL-3-dependent cell lines and T cell lines utilized the same transcriptional start sites. In chloramphenicol acetyltransferase (CAT) assays, the minimal 70-bp promoter confers strong transcriptional activity which is 50 60% of the Moloney long terminal repeat promoter activity. The 500-bp promoter region linked to the CAT gene exhibits IL-3 dependency similar to the endogenous TcR gamma gene. The immediate 3' and 5' flanking sequences inhibit the promoter activity two- to fourfold. The promoter lacks an obvious TATA box or CAAT box sequences, but contains a GC box in the untranslated region 3' to the promoter. The GC box is the core sequence of the element which binds Sp1-like proteins. Cloning of this Sp1 binding element in front of the thymidine kinase (TK) promoter and mutations generated in this site demonstrate its function as a silencer. Ultraviolet cross-linking analysis with the Sp1 binding site from the TcR gamma promoter revealed binding of a 90-100-kDa protein in a T cell line (EL 4) and 40-50 and 90-100-kDa proteins in FDC-P1 cells. The possible function of the Sp1-like protein in silencing the minimal promoter activity is discussed. PMID- 7489746 TI - Naive CD4+ T cells confer idiotype-specific tumor resistance in the absence of antibodies. AB - CD4+ T cells can recognize a processed idiotypic peptide derived from the mouse lambda 2(315) immunoglobulin light chain. The idiotypic peptide is presented on the I-E(d) class II major histocompatibility complex molecule. Mice made transgenic for a lambda 2(315)-specific alpha beta T cell receptor have been demonstrated to be specifically resistant against a tumor challenge with the MOPC315 (alpha,lambda 2(315)) plasmacytoma (Lauritzsen, G. F., Weiss, S., Dembic, Z. and Bogen, B., Proc. Natl. Acad. Sci. USA 1994, 91: 5700). That study, however, did not rule out a role of either anti-Id antibodies or T cells expressing nontransgenic specificities due to expression of endogenous T cell receptor (TcR) alpha chains. Also, the role of different T cell subsets in protection was unclear. To remove these ambiguities, we have now made the transgenic mice homozygous for the scid mutation, known to inhibit both Ig and TcR gene rearrangements. Such transgenic SCID mice lack B cells and antibodies while they still have plenty of CD4+ and CD4-8- cells expressing the transgenic alpha beta T cell receptor. The number of CD8+ T cell is dramatically reduced. Even so, transgenic SCID mice are protected against a challenge with MOPC315 plasmacytoma cells. Therefore, B cells, as well as novel T cell receptor specificities created by rearrangements of endogenous alpha-chain genes, are both dispensable for effective immunosurveillance in our system. Surprisingly, we found that transgenic CD8+ and CD4-8- cells are idiotype-specific and I-E(d) restricted. However, these T cell subsets are not required for resistance because adoptive transfer experiments demonstrated that highly purified transgenic SCID CD4+ cells suffice for tumor protection. PMID- 7489747 TI - CD28:B7 interactions promote T cell adhesion. AB - CD28 activation by antibody-mediated ligation has been shown to provide an important co-stimulatory signal for T cell adhesion to purified protein ligands. However, the effect of CD28 ligation by one of its natural ligands, B7.1, on T cell adhesion to other cells has not been studied. Therefore, in the present manuscript, we characterized the adhesive interactions between human T cells and B7.1-transfected major histocompatibility complex class II+ and class II- melanoma cells. In our studies, human T cells and T cell clones adhered to B7.1 transfected melanoma cells, but not to untransfected parental cells. The adhesive reaction in this model was rapid, occurring within 15 min, and was inhibited by anti-B7.1 antibody and soluble CTLA-4 immunoglobulin. Antibody inhibition studies demonstrated that adhesion between T cells and B7.1-transfected melanoma cells was mediated by interactions between LFA-1:ICAM-1 and CD2:LFA-3. Inhibition by pharmacological agents demonstrated that the CD28-induced adhesion required specific intracellular signaling events. A protein kinase C inhibitor, staurosporin, significantly inhibited T cell binding to transfected melanoma cells, while cyclosporin A and wortmannin, an inhibitor of phosphatidylinositol-3 kinase, did not. These results suggest that the presence of B7 on various cell populations may activate lymphocytes to adhere better, thus promoting activation, cytolysis, and migration. PMID- 7489749 TI - Dendritic cells generated from peripheral blood transfected with human tyrosinase induce specific T cell activation. AB - Peptides of melanosomal proteins have recently been shown to be recognized in an HLA-restricted mode by specific cytolytic T lymphocytes in melanoma patients. Dendritic antigen-presenting cells (DC) are considered to be the most effective stimulators of T cell responses, and the use of these cells has therefore been proposed to generate therapeutic responses to tumor antigens in cancer patients. We, therefore, generated DC from peripheral blood of normal donors in the presence of granulocyte/macrophage colony-stimulating factor and interleukin-4. Flow cytometric analysis of the cells during a 2-week culture revealed a loss of CD14 and CD34 expression, a concomittent increase of CD1a, CD11a,b and c, CD44, CD45, CD54, HLA-class I and II, and intermediate levels of CD26, CD80 and CD86. Cultured DC stimulated proliferation of allogeneic T cells and induced a marked, up to 20-fold, stimulation of T cell proliferation after pulsing with tetanus toxoid. To achieve independence of already-identified antigenic peptides presented in HLA class I-restricted fashion, which limits the general applicability of such peptides for vaccination of melanoma patients, we tested whether DC are transfectable with eukaryotic expression plasmids. DC transfected with two reporter genes (CAT, beta-galactosidase) using a liposome-based transfection technique, exhibited only low levels of enzymatically active proteins, but were able to degrade rapidly intracellular proteins and to process peptides efficiently. Chloramphenicol acetyltransferase as well as tyrosinase mRNA were detectable after transfection by reverse-transcriptase-polymerase chain reaction, and enzyme activities became measurable. Furthermore, DC transfected with the tyrosinase gene were able to induce specific T cell activation in vitro, indicating appropriate peptide processing and presentation in DC after transfection. These data suggest new approaches to future tumor vaccination strategies. PMID- 7489750 TI - Immature B cells from human and mouse bone marrow can change their surface light chain expression. AB - The capacity of bone marrow-derived surface immunoglobulin-positive (sIg+) human and mouse immature B cells, generated either in vitro or in vivo, to change their light (L) chain expression, has been assayed by the number of cells which change in vitro from one type of L chain to the other type, or to no sIg at all. Immature sIg+ B cells were generated in vitro from sIg- precursor cells from human or mouse bone marrow. The immature sIg+ cells expressed RAG-1. Human sIg+ cells expressed kappa and lambda L chains in ratios between 1:1 and 3:1, whereas in mouse cells, this ratio ranged from 10:1 to 20:1. Upon reculture of the human and mouse kappa+ sIg+ cells, about half of them remained kappa+, a quarter became lambda+, and another quarter became sIg-. Between 1 and 3% expressed both kappa and lambda chains. Of the human lambda+ cells, about two-thirds remained lambda+, only 1 to 2% became kappa+, while the other third became sIg-. Again, between 1 and 3% expressed both kappa and lambda L chains. These results indicate that expression of sIgM in the B cell membrane does not terminate L chain gene rearrangement, and that some order exists in kappa versus lambda gene rearrangements. Hence, human and mouse kappa+ immature B cells can become lambda+, but very few of the lambda+ cells can become kappa+, and both can become sIg-. Further, human CD10+/sIg+ kappa+ and lambda+ cells and mouse B220low/sIglow kappa+ cells enriched from bone marrow, i.e. immature B cells differentiated in vivo, changed their Ig phenotype upon in vitro culture, but in lower frequencies. By contrast, human and mouse mature B cells did not change their L chain or Ig phenotype. Hence, at least a part of the sIg+ immature B cells in bone marrow retain the capacity to change their L chain and Ig phenotype, and this capacity is lost when they become mature, peripheral B cells. PMID- 7489751 TI - Efficient immune responses in mice lacking N-region diversity. AB - Mice with a null mutation in the terminal deoxynucleotidyl transferase (TdT) gene harbor immunoglobulin and T cell receptor repertoires essentially devoid of N region diversity. Consequently, the CDR3 loops important for antigen recognition are shorter and considerably less diverse than those of wild-type controls. We find surprisingly normal immune responses in TdT0 mice, as regards both efficiency and specificity. This provokes a reconsideration of the assumption that N-region diversity is required for an effective T and B cell repertoire. PMID- 7489748 TI - Pertussis toxin-sensitive GTP-binding proteins regulate activation-induced apoptotic cell death of human natural killer cells. AB - Apoptosis of natural killer (NK) cells can be induced by non-specific physical damage (UV irradiation, heat shock) or by simultaneous ligation of the CD16 and the interleukin-2 receptor (IL-2R) molecules, but not with either anti-CD16 or IL 2 alone. Whereas blockade of GTP-binding protein (G protein)-mediated signal transduction using ADP-ribosylating bacterial toxins or the GTPase-resistant GTP analog guanosine 5'-0-(3-thiotriphosphate (GTP gamma S) does not affect non specific induction of NK cell apoptosis, such interventions do inhibit induction of apoptosis by anti-CD16/IL-2. The G proteins involved in the regulation of activation-induced NK apoptosis are sensitive to pertussis toxin (PTX) and to the non-specific GTP analog GTP gamma S but not to cholera toxin, Pseudomonas exotoxin A or diphtheria toxin. A pertussis toxin mutant that lacks ADP ribosylating activity, but conserves the membrane translocating and T cell mitogenic effects of the native molecule, fails to inhibit NK apoptosis. To exert their apoptosis-inhibitory effect, PTX and GTP gamma S must be employed before cells are activated. Later addition has no effect, suggesting the implication of G proteins in the transmission of apoptosis-inducing signals, but not in the effector stage of apoptosis. Pre-incubation with PTX or GTP gamma S does not affect the activation of NK cells by CD16 cross-linking, IL-2 stimulation- or both, as assessed by the induction of CD69 expression, protein tyrosine phosphorylation and calcium mobilization. Moreover, neither PTX nor GTP gamma S compromise the effector function of NK cells or the susceptibility of target cells to NK-mediated lysis. These data suggest apoptosis as a novel mechanism by which NK responses may be controlled in vivo, as well as an experimental and therapeutical strategy to counteract endogenous down-regulation of NK responses. PMID- 7489752 TI - Autoreactive T cells in healthy individuals show tolerance in vitro with characteristics similar to but distinct from clonal anergy. AB - Peripheral tolerance to self antigens has been suspected to play an important role in the regulation of the immune response in humans since autoreactive T cells can be isolated from the peripheral blood of healthy individuals. The mechanism of this tolerance is not known, but a number of groups have shown that autoreactive T cells can be induced to proliferate in vitro by the addition of their specific antigen and exogenous interleukin (IL)-2. In this report, we present the analysis of autoreactive T cells, isolated from healthy individuals, to the autoantigen GpIIb-IIIa present on circulating bone-marrow-derived cells and on thymic epithelial cells. We found that the response of GpIIb-IIIa autoreactive T cells in vitro, when stimulated with GpIIb-IIIa, shares characteristics with the response found for anergic T cells. In response to GpIIb IIIa, the GpIIb-IIIa-autoreactive T cells are neither able to proliferate nor produce IL-2 on their own, but do express IL-2 receptors alpha on their cell surface and produce IFN-gamma. This state of unresponsiveness can be broken by the addition of exogenous IL-2 and IL-7, as in the case of anergic T cells. However, GpIIb-IIIa-autoreactive T cells differ from anergic T cells in their capacity to be stimulated by IL-12 and by their production of IL-2 mRNA. Interestingly, once the unresponsive state to GpIIb-IIIa has been broken by the addition of IL-2, GpIIb-IIIa autoreactive T cells can produce IL-2 and proliferate when restimulated by GpIIb-IIIa alone. Altogether, these results suggest that the tolerance of GpIIb-IIIa autoreactive T cells from healthy individuals could involve post-transcriptional regulation of IL-2 expression. PMID- 7489753 TI - Natural antibodies against the immunoglobulin F(ab')2 fragment cause elimination of antigens recognized by the F(ab')2 from the circulation. AB - A humanized monoclonal IgG1 antibody, designated hC4G1, recognizes the fibrinogen receptor glycoprotein (GP)IIb/IIIa on platelets and inhibits platelet aggregation. When the F(ab')2 fragment of hC4G1 (F(ab')2 hC4G1) was administered to cynomolgus monkeys, all the monkeys showed inhibition of platelet aggregation ex vivo. Unexpectedly, a significant decrease in platelet count was observed in 5 of 18 monkeys. Antibodies against F(ab')2 hC4G1 were detected in the plasma of these monkeys by ELISA. Antibody activity in the plasma of these monkeys was significantly correlated with the intensity of platelet decrease (r = 0.84). The natural monkey antibodies to F(ab')2 hC4G1 were directed against the C-terminal region of F(ab')2 fragment common to all human and humanized IgG antibodies. Natural homo-reactive antibodies were also detected in human plasma from 15 of 40 healthy volunteers. Specificity was closely similar to that of the monkey antibodies. Affinity-purified human homoreactive antibodies enhanced phagocytosis of platelets treated with the F(ab')2 hC4G1. Monkey plasma with high homo reactive antibody activity was confirmed to decrease platelet count when administered together with F(ab')2 hC4G1 to a monkey with low antibody activity. These results suggest that F(ab')2 of humanized and human antibodies causes elimination of the corresponding antigens from the circulation by homo-reactive antibodies. PMID- 7489754 TI - Exacerbated autoimmunity associated with a T helper-1 cytokine profile shift in H 2E-transgenic mice. AB - Major histocompatibility complex (MHC) class II genes are the strongest susceptibility markers for many human autoimmune diseases. A perplexing aspect of this is that HLA alleles can confer either susceptibility or dominant protection. In nonobese diabetic (NOD) mice, the strongest known diabetes susceptibility locus is within the MHC and is presumed to be the H-2Ag7 product. When NOD mice carry a transgenic E alpha d molecule allowing expression of an H-2E heterodimer, diabetes is prevented. We investigated whether, as in human autoimmunity, a single class II heterodimer might protect from some autoimmune diseases while predisposing to others. NOD mice are susceptible to experimental autoimmune encephalomyelitis (EAE) induced by the proteolipoprotein (PLP) epitope 56-70. Susceptibility to EAE was analyzed in NOD mice which either have or lack transgenic H-2E expression. We found that H-2E expression in NOD mice has converse effects on diabetes and EAE: while diabetes is prevented, EAE is greatly exacerbated and leads to demyelination. Although PLP 56-70 could be presented both in the context of H-2A and H-2E, increased disease severity in H-2E transgenic mice could not be attributed either to an enhanced T cell proliferative response to PLP or to differences in determinant spread. However, cytokine analysis of the response revealed important differences between NOD mice and their H-2E transgenic counterparts: H-2E expression was associated with reduced interleukin-4 secretion and enhanced interferon-gamma (IFN-gamma) secretion by lymph node cells, while the response of central nervous system infiltrating T cells displayed a markedly enhanced IFN-gamma response. Thus, whether a particular class II molecule confers resistance or susceptibility to an autoimmune disease may depend on differential cytokine profiles elicited by particular class II/autoantigen complexes. PMID- 7489755 TI - The presence of the HLA class II allele DPB1*0501 in ethnic Thais correlates with an enhanced vaccine-induced antibody response to a malaria sporozoite antigen. AB - In this study, we examined the correlation between the frequency of allelic variants of the class II human leukocyte antigen (HLA) DR, DQ and DP gene loci and the quantitative humoral immune response observed in 71 Thai volunteers, subsequent to vaccination with a conjugated subunit vaccine. This vaccine was designed to induce antibodies directed against the immunodominant repeat region of the Plasmodium falciparum circumsporozoite (CS) protein. The presence of the DPB1*0501, a relatively common allele in Asian populations, was found to be associated with high vaccine-induced CS repeat-specific antibody responses in the volunteers. Given the increasing focus on the use of subunit vaccines in the control of infectious diseases, consideration of the influence of class II allele frequencies in ethnically diverse recipient populations may be important. PMID- 7489757 TI - Allo-skin graft rejection, tumor rejection and natural killer activity in mice lacking p56lck. AB - Mice lacking the p56lck molecule (lck -/-) have a profound block in the maturation of thymocytes and a greatly reduced number of peripheral mature T cells. To analyze further the functions of the T cells developed in lck -/- mice in vivo, we evaluated the ability of lck -/- mice to reject allo-skin grafts and methylcholanthrene (MCA)-induced syngeneic fibrosarcoma, and also examined the biological activity of lck -/- natural killer (NK) cells. Mice lacking p56lck failed to reject skin grafts from either MHC-disparate or minor histocompatibility-different donors, even after they had been primed with donor spleen cells. They also failed to reject the MCA-induced immunogenic syngeneic fibrosarcoma, MC57X. NK activity in mice lacking p56lck was normal, and there were no differences in the NK cell activation induced by poly(I).poly(C) stimulation or interleukin-2 stimulation (lymphokine-activated killer induction) between mice lacking p56lck and their immunocompetent heterozygous littermates. NK cells lacking p56lck mediated a normal antibody-dependent cell-mediated cytotoxicity (ADCC) response. The results of this study indicate that the loss of p56lck severely impairs the effectors of the immune system which mediate the rejection of allo-skin grafts and syngeneic tumors. The normal NK activity in lck -/- mice suggests that p56lck is not required for the development and activation of NK cells. PMID- 7489756 TI - Fas receptor expression on B-lineage cells. AB - Mice homozygous for the lpr mutation have B and T cell defects and develop autoantibodies, suggesting that lpr plays a role in their genesis. The lpr defect has been identified as a mutation in the apoptosis-associated Fas receptor (FasR) gene. To begin to define the role of FasR in B cells, we have surveyed FasR expression on B-lineage cells from early progenitors in the bone marrow through their maturation in the periphery. Contrary to some reports, we found that FasR is expressed on B cells at all stages of their development and is highest on germinal center B cells. FasR is not expressed on lpr!lpr-derived cells. These data are consistent with the idea that lpr/lpr mice have an intrinsic B cell defect that may be manifested in developing as well as peripheral B cells. An unexpected finding is that B-1 (CD5) B cells do not constitutively express FasR: FasR becomes detectable on B-1 B cells only after activation. PMID- 7489759 TI - Are murine marginal-zone macrophages the splenic white pulp analog of high endothelial venules? AB - The entry of lymphocytes into the spleen, in contrast to lymph nodes, does not involve high endothelial venule (HEV) interaction. The precise point of entry, as well as the mechanism by which lymphocytes enter the lymphoid areas of the spleen, remains controversial. We examined in detail the effect of two agents, pertussis toxin (PT) and the sulfated polysaccharide fucoidan, on splenic lymphocyte entry and positioning. These have previously been shown to interfere with lymphocyte extravasation across HEV. PT prevents lymphocyte extravasation, but not binding, to HEV, whereas fucoidan prevents binding and thus subsequent extravasation. Studies presented here show that pretreatment of murine lymphocytes with PT does not numerically affect entry into spleen, but profoundly alters lymphocyte positioning within the spleen. When fluorescently labeled, PT treated lymphocytes are injected intravenously, they initially accumulate in the marginal zone, in apparent association with the layer of marginal zone macrophages (MZM phi) which form a shell around the white pulp. They fail to traverse this layer into the white pulp, and subsequently localize in the red pulp. In contrast, untreated cells initially appear in the marginal zone, then continue to migrate into the white pulp after traversing the MZM phi layer. The localization of PT-pretreated lymphocytes adjacent to the MZM phi layer is disrupted by intravenous administration of fucoidan. Using a flow cytometric assay of aggregation between MZM phi and lymphocytes, we confirmed that fucoidan is also able to inhibit this association in vitro, whereas PT has no effect on this interaction. We propose that MZM phi in the mouse are the splenic analog of HEV, forming the port of entry of lymphocytes into the white pulp of the spleen. PMID- 7489758 TI - Engagement of CD20 suppresses apoptosis in germinal center B cells. AB - In a screen of 67 monoclonal antibodies (mAb) included in the Blind Panel of B cell antibodies for the 5th International Workshop on Human Leukocyte Differentiation Antigens, only the CD20 mAb--included as a positive control for immunophenotyping studies--was found to suppress the spontaneous apoptosis which occurs in human germinal center (GC) B cells when placed in tissue culture at 37 degrees C. Further detailed study using the 1F5 mAb confirmed this observation, showing that rescue from apoptosis via CD20, while not as efficient as that obtained on ligating CD40, was of similar magnitude to that achieved on engagement of surface immunoglobulin (sIg) by immobilized antibody. Also similar to anti-Ig, the CD20 mAb rescued from apoptosis without priming for the proliferation of GC B cells: this was quite different to its action on resting, non-GC B cells, where it provides a potent priming signal for cell cycle progression in response to IL-4 or anti-CD40. Unlike the survival signal engendered via sIg, CD20 engagement neither mobilized Ca2+ from intracellular stores or opening of a Ca2+ channel with 1F5, nor did it affect the ability of anti-Ig to open a Ca2+ gate in GC B cells. An unexpected feature of CD20-mediated rescue of GC B cells from apoptosis was a failure to turn on Bcl-2 expression. PMID- 7489760 TI - Cross-recognition by T cells of an epitope shared by two unrelated mycobacterial antigens. AB - The molecular mimicry represented by cross-recognition of determinants shared by unrelated antigens by antibodies or T cells is of broad immunological interest. In this study, we analyzed the cross-recognition by CD4+ T cells of a peptide epitope shared by two mycobacterial proteins of diverse sequence, represented by the 19-kDa antigen of Mycobacterium tuberculosis and the 28-kDa antigen of Mycobacterium leprae. This epitope was immunodominant with respect to the 19-kDa antigen, but cryptic in relation to the 28-kDa antigen. The cross-reactive epitope cores were identified by Pepscan window analysis and found to be eight residues long in both antigens (residues 69-76 and 127-134). Alignment of these octameric sequences revealed two identical and five conservatively related amino acids. Within the epitope core, two residues (73Asn and 76Ile) were identified as critical for recognition on the basis of inhibition of the cross-reactive T cell proliferative response using singly substituted analog peptides. These results suggest that T cell cross-reactive epitopes can exist in proteins with apparently not more than random levels of sequence homology. Their potential for unsuspected cross-sensitization may play a role in the maintenance of T cell memory, in the pathogenesis of autoimmune diseases and possibly in a wide range of host immune responses to infectious pathogens. PMID- 7489761 TI - Elucidation of the protein kinase C-dependent apoptosis pathway in distinct subsets of T lymphocytes in MRL-lpr/lpr mice. AB - MRL-lpr mice are severely impaired in the Fas pathway of apoptosis induction. We here evaluate another pathway of apoptosis induction in MRL-lpr mice which is protein kinase C (PKC) dependent. Despite the defect of the Fas pathway, apoptosis developed during culture in vitro in splenic T lymphocytes from MRL-lpr mice more extensively than in T lymphocytes from MRL-(+/+) mice. Apoptosis induction in the former cells was then found to be greatly promoted by PKC inhibitor H-7, and partially prevented by PKC activator phorbol 12-myristate 13 acetate (PMA). High sensitivity to H-7, but not to PKA inhibitor HA 1004, of these cells for apoptosis induction was confirmed by detailed time course and dose-dependency experiments of the drug effect. Population analysis showed that both CD4+ T lymphocytes and CD8+ T lymphocytes from MRL-lpr mice were highly sensitive to H-7, whereas CD8+ T lymphocytes, but not CD4+ T lymphocytes, from MRL-(+/+) mice were susceptible to the reagent. Interestingly, B220+ Thy-1+ CD4 CD8- T lymphocytes from MRL-lpr mice were most sensitive to H-7 for apoptosis induction. Correspondingly, the membrane-translocated activated PKC-alpha level in splenic T lymphocytes from MRL-lpr was more extensively up-regulated by PMA than in splenic T lymphocytes from MRL-(+/+). These results suggest that some signal consistently activates PKC in MRL-lpr T lymphocytes, and this event is needed for survival of these cells. On the other hand, CD4+ CD8+ thymocytes were deleted by apoptosis in culture with PMA, whether these thymocytes were from MRL lpr mice or MRL-(+/+) mice. This finding suggested that the apoptosis induction pathway linked to PKC activation is intact in CD4+ CD8+ thymocytes from the Fas defective MRL-lpr mice. We conclude from these results that the PKC-dependent signal pathways for either cell death or cell activation are intact or even accelerated in lpr mice, which could both compensate for the loss of the Fas pathway and promote the generation of autoreactive T lymphocytes. PMID- 7489762 TI - Bacterial superantigens induce T cell unresponsiveness in B cell-deficient mice. AB - Bacterial superantigens such as staphylococcal enterotoxin B (SEB) cause in vivo a profoud and long-lasting state of unresponsiveness in ligand-reactive T cells. To test whether presentation of SEB by small resting B cells to ligand-reactive T cells is essential for the induction of T cell unresponsiveness, we analyzed the effect of SEB in B cell-deficient mice. We observed T cell deletion and T cell unresponsiveness in both B cell-deficient mice and control mice. We conclude that presentation of SEB by resting B cells is not a prerequisite for the induction of T cell unresponsiveness in vivo. PMID- 7489763 TI - Der p I, a major allergen of the house dust mite, proteolytically cleaves the low affinity receptor for human IgE (CD23). AB - The nature of the proteases that cleave CD23 in vivo is of considerable interest, but remains unknown. Here, we demonstrate that Der p I, a major allergen of the house dust mite Dermatophagoides pteronyssinus, cleaves CD23 from the surface of cultured human B cells (RPMI 8866 B cell line). The cleavage of the receptor from the B cell surface was associated with a parallel increase in soluble CD23 (sCD23) in the culture supernatant. Furthermore, the proteolytic effect of Der p I was specific for CD23, since none of the other B cell markers tested (CD20, HLA DR, CD71 and CD49d) were affected. Labeled antibody experiments and protease inhibition assays clearly demonstrate that Der p I is a cysteine protease that directly cleaves a 25-kDa fragment of CD23. These data suggest that the cysteine protease Der p I, in addition to being highly immunogenic, may up-regulate IgE synthesis by virtue of its ability to cleave CD23. PMID- 7489764 TI - Differential interactions between the immunoglobulin heavy chain mu intron and 3' enhancer. AB - The immunoglobulin heavy chain 3' enhancer may be a novel type of a transcriptional regulation element in as much as its function is position dependent. We show that there are interactions between the mu intron and 3' enhancer which are differentially regulated depending on the distance between the two elements. Thus, a transcriptional repression is exerted by the 3' enhancer when juxtaposed to the intron enhancer. Whereas no or only modest synergism between the immunoglobulin mu intron and 3' enhancer has been reported to date, we show here that the stimulatory effect is substantially increased by extending the distance between the two enhancers. In our stable expression system, the mu intron enhancer insulated the test gene from neighboring chromatin. PMID- 7489765 TI - Relevance of residue 116 of HLA-B27 in determining susceptibility to ankylosing spondylitis. AB - Ankylosing spondylitis (AS) is an autoimmune disorder strongly associated with HLA-B27. A direct role of B27 molecules in the disease pathogenesis has been postulated, possibly by presenting to T cells an as-yet unidentified arthritogenic peptide that triggers the autoimmune response. There are nine HLA B27 alleles differing from each other at one or more amino acid positions. It is important, for the identification of the arthritogenic peptide, to define which alleles, and therefore which polymorphic positions, predispose to the disease. Here, we report that HLA-B*2709 is not associated with AS, as it was not found in patients. HLA-B*2709 differs from the most frequent and disease-associated HLA B*2705 allele for a single substitution (His vs. Asp) at position 116. Amino acid 116 is located at the bottom of the groove where the antigenic peptide sits, and it has been proven to influence the peptide-binding specificity of HLA class I molecules. The most likely interpretation of these data is that the differences in charge and size that accompany the His-to-Asp substitution exclude the acceptance of the arthritogenic peptide. PMID- 7489766 TI - Tumor necrosis factor-alpha induction of major histocompatibility complex class II antigen expression is inhibited by interferon-gamma in a monocytic cell line. AB - Regulation of major histocompatibility complex (MHC) class II antigen expression by cytokines has been suggested to play a major role in the initiation and propagation of immune and autoimmune processes. The analysis of class II gene regulation benefits greatly from the existence of mutants with defects in regulatory factors. We report the establishment of a subclone of the human monocytic cell line U937, termed C119/9, with unusual cytokine regulation of MHC class II expression. In contrast to the parental U937 cell line, only tumor necrosis factor (TNF)-alpha, and not interferon (IFN)-gamma induces the expression of MHC class II antigens on C119/9 cells, and paradoxically, this induction was inhibited almost completely by IFN-gamma. The HLA-DR induction is controlled at the transcriptional level by the first 150 bp of the class II promoter which contains all the class II consensus elements. Both HLA-DR and -DQ mRNA are induced by TNF-alpha treatment, and both are diminished upon co treatment with TNF-alpha and IFN-gamma. This antagonism between TNF-alpha and IFN gamma seem to be restricted to MHC class II genes. This subline of U937 cells may be useful in further studies of MHC class II regulation. PMID- 7489767 TI - Pollution of modern metalworking fluids containing biocides by pathogenic bacteria in France. Reexamination of chemical treatments accuracy. AB - Pollution by pathogenic bacteria was examined in 150 French metalworking fluid samples. Gram-negative micro-organisms such as Salmonella spp., Shigella spp., and Vibrio spp. as well as Gram-positive cocci were never isolated. Nevertheless opportunistic pathogens such as Pseudomonas aeruginosa and Klebsiella pneumoniae still contaminated these fluids with an isolation frequency of 17% of samples for each. These two micro-organisms failed to grow or even survive in vitro in sterile cutting fluids protected by biocides. Preliminary growth of other micro organisms such as Pseudomonas putida or Pseudomonas fluorescens, which are the major part of the indigenous microflora, seemed to be a prerequisite for their growth. These former two Pseudomonas could resist three different classes of biocides and, at least in the case of formaldehyde-releasers, adaptation was followed by biocide deterioration. Resistance magnification was observed in the presence of the three different types of biocides and, in the case of formaldehyde releasers the resistance and deterioration levels were close to those recommended by the manufacturers. This is probably the reason why the preliminary growth of Pseudomonas putida allowed in vitro differed growth of Klebsiella pneumoniae and Pseudomonas aeruginosa. Due to relatively high isolation frequencies of opportunistic pathogens (17% of samples) periodical microbiological examination of cutting fluids should be carried out in order to evaluate risks for human health. Wearing masks and gloves is still recommended, at least in France. PMID- 7489768 TI - Diphtheria: changing patterns in the developing world and the industrialized world. AB - In the past, diphtheria was considered one of the most serious childhood diseases because it took a heavy toll in health and life among preschool-aged children. Prior to the widespread availability of diphtheria toxoid, nearly 70% of cases were in children younger than 15 years of age. In the industrialized countries, immunization against diphtheria became widespread in the 1940s and 1950s. This led to a marked decrease in the incidence of diphtheria. There was also a decrease in circulating toxigenic Corynebacterium diphtheriae organisms, resulting in less natural boosting of antibody levels. This had led to gaps in the immunity of the adult population. Since 1990, diphtheria has made a spectacular comeback in several European countries, with a high proportion of cases in adults. In developing countries, immunization of infants with diphtheria toxoid was introduced with the Expanded Programme on Immunization in the late 1970s. Coverage rose slowly to 46% in 1985 and 79% in 1992. Because the pool of immunized persons is not yet large, the process of maintaining immunity still operates through natural mechanisms, including frequent skin infections caused by C. diphtheriae. But recently, several developing countries where coverage has been high for 5-10 years have reported diphtheria outbreaks. These outbreaks have been characterized by high case fatality rates, a large proportion of patients with complications, and their occurrence in both young and older age groups. In all countries, priority should be given to efforts to reach at least 90% coverage with three doses of diphtheria toxoid in children below one year of age. In countries where diphtheria has been successfully controlled, immunity levels should be maintained by booster doses. PMID- 7489769 TI - Impact of nitrates in drinking water on cancer mortality in Valencia, Spain. AB - The concentrations of nitrates in public drinking water in the Mediterranean coastal province of Valencia are not only the highest in Spain but also in the whole of Europe. Intensive agricultural practices involve a traditional and growing use of nitrogen fertilizers. This and the terrain--poorly consolidated and porous in areas--favors the accumulation of nitrates in underground aquifers, thereby perhaps accounting for this contamination. The possible conversion of nitrates to nitrites under certain conditions of gastric achlorhydria, followed by their transformation to nitrosamines--substances known to be carcinogenic in experimental models--has led to a number of epidemiological studies of the possible relationship between high nitrate levels in public drinking water and mortality due to different cancers. The aim of the present study was to analyze the relationship between different levels of exposure to nitrates in the drinking water of the 258 municipalities in the province of Valencia and mortality due to cancer of the stomach, bladder, prostate and colon in this population. The cancer mortality rate was found to rise with increasing exposure to nitrates in the case of gastric cancer in both sexes, and in prostate cancer. These same results were obtained on calculating relative risk for the different age groups associated with the consumption of drinking water containing different levels of nitrates.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7489770 TI - AIDS related information, attitudes and behaviours among Italian male young people. AB - In order to evaluate the AIDS related sources of information, knowledge, attitudes and behaviours among male young people it was filled in a multi-choice anonymous questionnaire by 2018 subjects, called for the first medical examination to Italian Navy and by 1348 recruits. The data from the latter group were used to make comparisons with our previous survey carried out in 1988. The most important sources of information on AIDS were mass media and school, on sexuality friends and family. AIDS awareness was greater among the high educated subjects and positively influenced by mass media and school as sources of information. In agreement with these data, knowledge was better about AIDS than about sexuality. Risk perception was positively influenced by the AIDS-related knowledge for called youth and a greater intolerance towards infected people was observed among the less educated and the less AIDS-aware subjects. On average 46% of the called up young men and 74% of recruits declared to be sexually active: among these, the mean age at the first sexual intercourse was 16, over 60% declared to have had multiple partners, over 30% unknown partners and over 30% didn't use condoms; 20% of called up and 38% of recruits indicated 2 or 3 risk behaviours. Factors determining sex-related risks were different according to the behaviour and the considered group. These results suggest that it is important for young people to be well informed about sex and AIDS before the beginning of sexual activity, if possible during compulsory school. PMID- 7489771 TI - Molecular characterization of trimethoprim resistance in salmonellas isolated in Sicily, 1985-1988. AB - The occurrence of trimethoprim (Tp) resistance in salmonellas isolated from humans and water samples in Sicily between 1985 and 1988 has been investigated and the Tp resistance mechanisms have been further characterized on the basis of hybridization with probes for the dihydrofolate reductase (DHFR) genes types I, II, IV and V. Of 765 strains examined, high level (> 1000 mg/l) resistance to Tp was identified in 23 strains (3%). In 22 of these strains, such resistance was associated with resistance to sulphonamides. Six serovars with Tp-resistant strains were identified, Salmonella typhimurium (14 strains), S. enteridis (2), S. agona (2), S. mbandaka (2), S. virchow (2), S. indiana (1). In all strains with high level Tp resistance, resistance to this antimicrobial was plasmid encoded, in most strains by plasmids with MWs ranging from 70-100 MDa. On the basis of restriction endonuclease analysis, four different categories of Tp resistance plasmids were identified in Tp-resistant strains of S. typhimurium. Hybridization with the DHFR I probe was observed in three strains of Tp-resistant S. typhimurium and two strains of Tp-resistant S. enteritidis; in contrast, in none of the strains tested was there any detectable hybridization with the probes for DHFR types II, IV and V. It is concluded that the DHFR type I resistance mechanism, common in Tp-resistant enterobacteria in many European countries, is relatively uncommon in Tp-resistant salmonellas isolated in Sicily. Furthermore, the DHFR V resistance mechanism, previously identified in strains of Shigella sonnei isolated in Sicily and associated with travellers from Sri Lanka, has not yet appeared in salmonellas in Sicily. PMID- 7489772 TI - HBsAg subtype distribution among different populations of HBsAg carriers in Spain. AB - Data concerning the HBsAg subtype distribution in Spain are out-of-date and confined to a restricted geographical area. Furthermore, the complex distribution observed in the countries surrounding Spain prevents any prediction. To obtain further data on HBsAg subtype distribution among Spanish HBsAg carriers, subtyping analysis (d and y determinants) was performed in 670 samples from subjects belonging to various epidemiological risk groups and coming from different geographical areas of the country. Similar frequencies were found for both mutually exclusive d/y subtype determinants among non-risk, normal HBsAg carriers from almost all geographical areas studied. In contrast, the ay subtype was clearly predominant (79-87%) among intravenous drug users, irrespective of their geographical origin. Thirteen different institutions for mentally retarded patients behaved as closed communities for HBV circulation, showing independent subtype distributions. Thus, no significant geographical variations were found for HBsAg subtype distribution in Spain. The prevalence of each particular subtype is mainly dependent on the epidemiological characteristics of the carriers studied. Subtype distribution was independent of the presence of HBeAg or HDV infection serum markers when homogeneous groups were considered separately. Atypical HBsAg phenotypes, either with coexistence or absence of both subtype determinants, were found in some cases. PMID- 7489773 TI - Seroepidemiology of Q fever in Nova Scotia: evidence for age dependent cohorts and geographical distribution. AB - To determine the incidence of Q fever in Nova Scotia, a randomly selected sample of 492 volunteers aged 18-70 years was recruited by mail from all 18 urban and rural counties in this province. Volunteers were followed from 1988 to 1991 for antibody titres to Coxiella burnetii antigens. Analysis of seroprevalence by age revealed two statistically different cohorts. Those younger than 35 years had a low, stable seroprevalence while those 35 years and older had a seroprevalence statistically correlated with increasing age. The finding of age related cohorts suggests a sporadic or cyclical infection rate. An age-independent geographical clustering was also noted, with higher seroprevalence in a major agricultural area of the province. No seroconversions were observed, but four subjects had seroreversions. At baseline, 72 (14.6%) of the 492 subjects were considered seropositive (> or = 1:8 C. burnetti phase II antibody titre by microimmunofluorescence). Of 47 seropositive subjects who provided yearly serum samples, two (4.3%) showed > or = 4-fold increase in antibody titre and did not react to antigens of the 11 other respiratory pathogens tested, suggesting a reactivation of Q fever. All 22 subjects positive to phase I antigen were positive to phase II antigen. This study suggests that the rate of infection in Nova Scotia has been low since the 1960's and that risk of infection is associated more with a geographically clustered reservoir of infection than with occupation or gender. PMID- 7489774 TI - A case-control study of gastric cancer and nutritional factors in Marseille, France. AB - A case-control study was conducted in Marseille (France) to investigate the relationship between usual diet and risk of gastric cancer. Patients with histologically confirmed gastric adenocarcinoma were identified in 8 major centres for gastric surgery. Controls were selected in specialized medical centres from patients undergoing functional reeducation for injuries or trauma, according to the age and sex distributions of the cases. The study involved 92 cases and 128 controls who were interviewed with a dietary history questionnaire on their usual diet during the year preceding first symptoms for cases, or preceding interview for controls. Odds ratios for specific foods were calculated after adjustment for age, sex, occupation and energy intake. A reduced risk was observed for consumption of raw vegetables (OR2: 0.55; OR3: 0.41 for the second and third tertiles, respectively), fresh fruit (OR2: 0.63; OR3: 0.50), vegetable oil (OR2: 0.60; OR3: 0.52), pasta and rice (OR2: 1.06; OR3: 0.50) whereas consumption of cakes and pastries (OR2: 1.02; OR3: 2.96), sugar and confectionery (OR2: 0.96; OR3: 1.68) was associated with an increased risk. An increased risk was found for intake of saturated fat (OR2: 1.49; OR3: 1.67), simple sugars (OR2: 1.18; OR3: 1.78) and calcium (OR2: 1.84; OR3: 2.57). A decreased risk was observed with intake of fiber (OR2: 0.49; OR3: 0.59), fibre from vegetables and fruit (OR2: 0.83; OR3: 0.53) and iron (OR2: 0.70; OR3: 0.41). PMID- 7489775 TI - Nitrosamine, nitrate and nitrite in relation to gastric cancer: a case-control study in Marseille, France. AB - A case-control study on gastric cancer and diet was conducted in Marseille (France). Ninety-two patients with histologically confirmed adenocarcinoma and 128 controls undergoing functional reeducation for injuries or trauma were interviewed by a trained dietician using a dietary history questionnaire on their usual diet during the year preceding the first symptoms for cases, or preceding interview for controls. Intake of nitrite, nitrate and pre-formed N nitrosodimethylamine (NDMA) from food was estimated using a food composition table compiled ad hoc. Odds ratios (ORs) were calculated after adjustment for age, sex, occupation and calorie intake. The results indicated that high intake of NDMA was associated with increased risk for gastric cancer. The ORs for the second and third tertile of NDMA intake were: OR2 = 4.13 (95% CI = 0.93 18.27) and OR3 = 7.00 (95% CI = 1.85 to 26.46). Intake of nitrate and nitrite was not associated with increased risk of stomach cancer. Consumption of vegetables was protective in general and independent of their estimated nitrate content. PMID- 7489776 TI - Effects of coffee on serum cholesterol and lipoproteins: the Italian brewing method. Italian Group for the Study of Atherosclerosis and Dismetabolic Diseases, Rome II Center. AB - In order to evaluate the effects of Italian brewed coffee (moka) on cholesterol and serum lipoproteins, a randomized double-blind 14-week clinical trial was performed. After a coffee-free period of four weeks, 49 subjects drank coffee, caffeinated and decaffeinated, for ten weeks. There was no evidence that the Italian method of brewing coffee affects serum lipoproteins since no statistically significant differences were found. PMID- 7489777 TI - Probable Creutzfeldt-Jakob disease after a cadaveric dural graft. AB - A 26-year old female who had received a cervical cadaveric dural graft 7 years earlier developed psychotic behavior and generalized myoclonic jerks. Following rapid deterioration and appearance of high-charge sharp periodic waves on electroencephalography she died within two months. We believe that this patient is the first German who acquired Creutzfeldt-Jakob disease by dural implant. PMID- 7489778 TI - Diffusion of thermophilic Campylobacter in the Pesaro-Urbino area (Italy) from 1985 to 1992. AB - The results of research on the spreading of campylobacter in the Pesaro-Urbino area carried out from 1985 to 1992 are presented. Materials of different origin were examined: 822 samples of human faeces, 533 animal rectal swabs, 192 samples of domestic sewage, 48 of river water, 96 of sea water and 632 of various types of food. Two hundred and nine strains of campylobacter were isolated (9%), most of which were Campylobacter jejuni (80%), with particular frequency in food products (chicken carcass 45.7%, ground meat and sausage 18.1%) and in river water (31.3%). In contrast, the samples of sea water and dairy cheese products were always negative. It may be concluded that the spreading of campylobacter in the Pesaro-Urbino area is mainly associated with food products of animal origin. Therefore, better controls in the processing of these products may be necessary. PMID- 7489779 TI - Helmet use by drivers and passengers of motorcycles in Pamplona (Spain), 1992. AB - In 1992 a new Traffic Rules Code was enacted in Spain. This Code included a helmet law that came into force on 1 September 1992. Since then, helmet use is compulsory for motorcycles in urban areas. Previously, helmet use had been obligatory only for motorcycle in highways. To evaluate driver and passenger compliance with the helmet law, an observational prevalence survey of helmet use in Pamplona (a city of 183,539 inhabitants in the north of Spain) was performed. The utilization of helmet by motorcycle drivers and their passengers was recorded three months before and three after the helmet law took effect. Helmet use increased from 19.7% in the first period to 94.8% in the second period. After the law took effect helmet use was higher among drivers 97.5% than among passengers 77.5%. Passengers used helmet more frequently when the driver was also using helmet. The prevalence ratio of helmet use, in those passengers whose driver used helmet versus those who didn't, was 9.91 (95% CI = 1.52, 64.83). Health education and law enforcement efforts should be targeted to drivers to prevent them from giving a ride to passengers without helmet. PMID- 7489781 TI - Comparison of the clinical and serologic diagnosis of haemorrhagic fever with renal syndrome (HFRS) and leptospirosis. PMID- 7489780 TI - Alcohol intake as a risk factor for peripheral arterial disease in the general population in the Edinburgh Artery Study. AB - The aim of this study was to determine the relationship between alcohol consumption and occurrence of peripheral arterial disease in the general population. During 1988 in a cross sectional survey, the Edinburgh Artery Study, 1,592 men and women aged 55-74 years were selected at random from the age-sex registers of ten general practices distributed geographically and socio economically across the city. participants were asked to recall the number of units of wine, beer and spirits consumed in the previous week and whether or not this was typical. Peripheral arterial disease was measured using the ankle brachial pressure index (ABPI). Men and women were analysed separately because of large differences in alcohol consumption. There was no association between ABPI and alcohol consumption in women but, in men, increasing alcohol consumption was associated with a higher ABPI (test for trend, p = 0.03) indicating less severe disease. This relationship was linear rather than U-shaped. In multiple regression analysis, after age-adjustment the ABPI was related to wine consumption but not beer or spirits in men (p < or = 0.01). On adjusting for age and cumulative lifetime cigarette smoking, the association of wine consumption with the ABPI was diminished but remained statistically significant (p < 0.05). On adjusting for age and social class, the relationship of total alcohol intake and wine consumption with the ABPI became non significant (p > 0.05). We conclude that in males, greater alcohol consumption is related to a higher ABPI and that any protective 'effect' of alcohol relates to wine consumption rather than beer or spirits.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7489782 TI - High prevalence of human T-lymphotropic virus (HTLV) infection in pregnant women in southern France. PMID- 7489785 TI - Growing consensus in COPD? PMID- 7489784 TI - The dark side of antituberculosis therapy: adverse events involving liver function. PMID- 7489783 TI - Resurgence of diphtheria. AB - Following the introduction of routine immunization with diphtheria toxoid in the 1940s and 1950s, diphtheria incidence declined dramatically in countries of the industrialized world. At the beginning of the 1980s many of these countries were progressing toward elimination of the disease. However, since the mid-1980s there has been a striking resurgence of diphtheria in several countries of Eastern Europe. For 1993, WHO received reports of 15,211 diphtheria cases in Russia and 2,987 cases in Ukraine. The main reasons for the return of diphtheria in these countries were: decreasing immunization coverage among infants and children waning immunity to diphtheria in adults, movements of the population during the last few years, and an irregular supply of vaccines. The outbreak spread to neighboring countries and in 1993 cases were reported in Azerbaijan, Belarus, Estonia, Finland, Kazakhstan, Latvia, Lithuania, Poland, Tajikistan, Turkey, and Uzbekistan. Epidemiological patterns of diphtheria are changing in developing countries, and the disease seems to be following patterns seen in industrialized countries 30 to 40 years ago. In developing countries, routine immunization against diphtheria was introduced in the late 1970s with the Expanded Programme on Immunization. In these countries, coverage of infants with 3 doses of diphtheria toxoid reached 46% in 1985, and 79% in 1992. Recent diphtheria outbreaks in Algeria, China, Ecuator, Jordan, Lesotho and Sudan demonstrate a shift in the age distribution of cases to older children and adults. Rapid clinical and public health responses are required to control diphtheria outbreaks. Three major measures are indicated: high immunization coverage of target groups, prompt diagnosis and management of diphtheria cases, and rapid identification of close contacts with their effective management to prevent secondary cases. PMID- 7489786 TI - Validation of the surveillance system for new cases of tuberculosis in a province of northern Italy. Varese Tuberculosis Study Group. AB - In Italy tuberculosis (TB) surveillance is hampered by several problems (inconsistency of data and underreporting). A tuberculosis surveillance system was established in Varese Province (VSS) in 1992. The aim of the present study was, using VSS, to estimate: 1) coverage and validity of data collected by the national Compulsory Surveillance System (CSS); 2) validity of diagnosis and risk factors for tuberculosis; 3) tuberculosis incidence. A specifically designed form, including the same items as the CSS form and 10 more questions on risk factors and diagnostic issue, was completed in all the existing health facilities in the province for each new tuberculosis case diagnosed, collected by the study co-ordinator during monthly supervision after a review of clinical records, and stored in a database at the co-ordinating centre. Routine notifications collected at national level by CSS for the province were nominally linked with VSS data. To evaluate coverage and data validity, a comparison was made between VSS and CSS data after removal of duplicates. Quality of data, risk factors and incidence were evaluated on VSS data (gold standard). CSS coverage was 63% with a significant underreporting of extrapulmonary cases. Within CSS, 20% of data were missing (particularly diagnostic information: 42-72%). According to VSS, smear was performed on 88% of cases and culture on 66%. Half of the cases had no risk factors. The overall incidence of tuberculosis in Varese Province was 15 per 100,000. We estimated the coverage and validity of the data collected by CSS, validity of diagnosis, risk factors for and incidence of tuberculosis and proposed several suggestions to improve CSS nationwide. PMID- 7489787 TI - A pseudoepidemic of Mycobacterium chelonae infection caused by contamination of a fibreoptic bronchoscope suction channel. AB - An unusual increase in the frequency of isolation of Mycobacterium chelonae subspecies chelonae from specimens of bronchial washings was found between September and December 1992 in National Taiwan University Hospital. During this period, a total of 123 patients underwent fibreoptic bronchoscopy with an Olympus P20. Seventy six patients had bronchial washing for bacteriological study and cytological examination. Acid-fast bacilli were found in 21 patients, in 18 of whom Mycobacterium chelonae were isolated from bronchial washing cultures. Eight patients were treated as mycobacterial infected, because of the presence of unexplained pulmonary lesion, positive acid-fast stain and culture for Mycobacterium chelonae. Diagnosis of lung cancer was delayed in one patient because of the initial negative cytological study and positive bacterial culture. The fibreoptic bronchoscope was disinfected by automated washing machine (EW-20, Olympus) using 2.3% glutaraldehyde according to a standard protocol. From a survey to search for possible sources of contamination, they were identified at the suction channel of four different bronchoscopes. This episode proved to be a pseudoepidemic. The contamination was controlled by extensive suction and rinsing of the channel with 70% alcohol immediately after disinfection by the automated bronchoscope disinfection machine. This study shows that, despite using the disinfection machine, the suction channel could still be contaminated with Mycobacterium chelonae. This may cause diagnostic confusion and unnecessary antimycobacterial treatment. PMID- 7489789 TI - Decreased lung function in one year survivors of allogeneic bone marrow transplantation conditioned with high-dose busulphan and cyclophosphamide. AB - Conditioning with busulphan (BU) and cyclophosphamide (CY) prior to allogeneic bone marrow transplantation (BMT) is an alternative to regimens that include total body irradiation (TBI). The aim of the study was to assess the occurrence and degree of lung function impairment after this treatment. Prospectively, 43 consecutive patients, aged 17-51 (median 31) yrs, were examined by lung function measurements and clinical and radiographic evaluation, prior to BMT and at 3 month intervals up to 1 yr after BMT. All patients had normal chest radiographs before BMT and at the 12 month follow-up. Mean baseline values were above 100% of predicted normal for lung volumes and above 90% for gas transfer. Excluded from the lung function follow-up analyses were nine patients who had suffered infectious pneumonia and/or developed obliterative bronchiolitis. For the remaining patients (n = 34), mean alveolar volume (VA), forced vital capacity (FVC) and forced expiratory volume in one second (FEV1) had dropped by nearly 10% compared with baseline 3 months after BMT, but were restored within 1 yr. FEV1/FVC x 100 (FEV1%) was increased, reflecting the restrictive pattern. Hb adjusted transfer factor of the lungs for carbon monoxide (TL,CO) had dropped by 20% after 3 months, and remained reduced by 15% after 1 year. Prior to BMT the smokers had significantly lower TL,CO than the nonsmokers, and after BMT the difference was accentuated. Reductions in lung function were independent of sex, age and type of haematological disorder. We conclude that BMT with BU/CY is associated with transient declines in lung volumes and a persistent reduction in gas transfer 1 yr after therapy.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7489788 TI - Deposition of amphotericin B aerosols in pulmonary aspergilloma. AB - The aim of the present study was to characterize amphotericin B aerosols nebulized by ultrasonic and jet nebulizers and to study their deposition and pharmacokinetics in patients with pulmonary mycetoma. The aerodynamic behaviour and pulmonary deposition of amphotericin B particles were measured using a direct isotopic method based on stable labelling of the drug with 99mTc. Each nebulizer was bench tested for inhaled mass and particle size distribution. Three patients suffering from pulmonary aspergilloma were enrolled for a 4 week clinical study. They received 5 mg of amphotericin B daily delivered by either Fisoneb or DP100 (ultrasonic) or Respirgard II (jet) nebulizers. Deposition of radiolabelled amphotericin B was measured once with each nebulizer using a gamma-camera. In two patients, amphotericin B serum concentration was monitored over a 330 min period after the nebulization had been completed. Inhaled masses of the three nebulizers, assessed as % of labelled drug caught in inspiratory filter in duplicate experiments, were: 5.8 and 3.6% for Respirgard II; 26.5 and 28.3% with Fisoneb; 5.9 and 6.3% for DP100. Mass median aerodynamic diameter (mean +/- SD) results were: 0.28 +/- 0.04 micron with Respirgard II; 4.82 +/- 0.78 microns with Fisoneb; and 2.27 +/- 1.14 microns with DP100. Because of larger particles and significantly greater inhaled mass, Fisoneb delivered more amphotericin B to the central airways, the lung periphery and in the mycetoma lung regions. Amphotericin B serum concentrations correlated with pulmonary deposition and remained below 25 ng.mL-1. No untoward effects were reported by the patients during the 4 week trial.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7489790 TI - Role of bronchoalveolar lavage in the diagnosis of fat embolism syndrome. AB - Fat embolism syndrome (FES) is a serious clinical disorder occurring in trauma patients. The diagnosis of fat embolism syndrome may be difficult to establish clinically. We therefore wanted to investigate the usefulness of bronchoalveolar lavage (BAL) in the diagnostic evaluation of fat embolism syndrome. We analysed the presence of fat droplets in BAL cells in 32 trauma patients (7 with full diagnostic criteria of fat embolism syndrome, 17 with incomplete diagnostic criteria, and 8 with no diagnostic criteria at the time of bronchoscopy), 9 nontrauma patients with acute respiratory failure and radiographic pulmonary infiltrates, and a control group composed of 10 individuals. An increased percentage of oil red O positive alveolar macrophages (cut-off point > 3%) in BAL was found in 6 out of 7 patients with definite clinical criteria of fat embolism syndrome, and in 6 out of 20 trauma patients without the clinical diagnosis of fat embolism syndrome. In two patients with fat embolism syndrome, sequential BAL showed that the percentage of positive macrophages decreased when the clinical manifestations disappeared. An increased number of BAL macrophages with fat droplets was also observed in two trauma patients without evidence of fat embolism syndrome after long-bone surgical intervention. By contrast, all non trauma patients had a percentage of positive cells lower than 3%. Our findings suggest that BAL oil red O positive macrophages are frequently observed in trauma patients irrespective of the presence of fat embolism syndrome. Conceivably, a high number of oil red O positive macrophages could reflect clinically silent fat embolization. PMID- 7489791 TI - Activated and memory alveolar T-lymphocytes in idiopathic eosinophilic pneumonia. AB - To investigate the possible involvement of T-lymphocytes in the immunopathogenesis of idiopathic eosinophilic pneumonia (IEP), we have evaluated the phenotypic characteristics both of peripheral blood and alveolar lymphocytes in six patients with symptomatic IEP, and in 24 healthy nonsmokers as controls, by employing bronchoalveolar lavage (BAL), monoclonal antibodies, and flow cytometry. In IEP, total and differential cell counts showed a mild alveolitis with an increase of eosinophil percentage and number; the alveolar lymphocyte count was also increased. In BAL, the total number both of CD4+ and CD8+ lymphocytes was significantly raised; CD4+ cells expressed early (CD25) activation antigens. The analysis of CD45R0, CD45RA and CD62L coexpression in IEP patients, when compared to healthy controls, revealed an accumulation of alveolar CD4+ cells showing phenotypic repertoire usually expressed by memory T-cells (CD45R0+, CD45RA-, CD62L-). CD8+ alveolar lymphocytes did not show any significant increase of activation antigen coexpression. Circulating lymphocytes were not significantly increased and showed only a significantly higher CD25 expression. These data suggest that a pivotal role is played by activated and memory CD4+ alveolar lymphocytes in IEP patients. PMID- 7489792 TI - Increased oxidation of extracellular glutathione by bronchoalveolar inflammatory cells in diffuse fibrosing alveolitis. AB - An unbalanced oxidative stress is thought to be an important element in the pathogenesis of diffuse fibrosing alveolitis (DFA). The purpose of our study was to investigate the role of reactive oxygen metabolites (ROMs) released from cultured bronchoalveolar inflammatory cells (BA-cells) on glutathione oxidation. We studied bronchoalveolar lavage samples from 10 healthy controls and from 20 patients with diffuse fibrosing alveolitis (all were nonsmokers). BA-cells obtained by bronchoalveolar lavage (BAL) were incubated with 50 microM of reduced glutathione (GSH). Oxidation of GSH to glutathione disulphide (GSSG) by BA-cell derived oxidants was detected as a decline of GSH in the supernatants. Total glutathione (GSHtot = GSH + 2 GSSG) and GSSG in the epithelial lining fluid (ELF), and methionine sulphoxide (Met(O)) content of BAL proteins were determined. In diffuse fibrosing alveolitis the oxidative activity of BA-cells was enhanced, GSHtot and GSH were decreased, whereas the GSSG:GSH ratio was increased. The oxidative activity of BA-cells correlated positively with the GSSG:GSH ratio, but not with the methionine sulphoxide content. The methionine sulphoxide content was elevated in diffuse fibrosing alveolitis and inversely correlated with GSHtot. The methionine sulphoxide content also correlated positively with the percentage of BAL neutrophils. We conclude that BA-cell derived reactive oxygen species are capable of oxidizing extracellular GSH in vitro. The positive correlation between the BA-cell oxidative activity in vitro and GSSG:GSH ratio in ELF suggests that a similar oxidative effect on extracellular GSH may also occur in vivo.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7489793 TI - A specific neutrophil elastase inhibitor (ONO-5046.Na) attenuates LPS-induced acute lung inflammation in the hamster. AB - We have examined the effect of ONO-5046.Na, a synthetic specific inhibitor of neutrophil elastase, on lipopolysaccharide (LPS)-induced acute lung inflammation. Syrian golden hamsters were injected intraperitoneally with either 300 mg.kg-1 of ONO-5046.Na or saline, 30 min before and 1 h after intratracheal administration of 0.1 mg.kg-1 LPS. Animals were sacrificed 2 and 24 h later and the wet-to-dry lung weight ratio (W/D) was determined. Bronchoalveolar lavage (BAL) was performed, and tissue sections were examined histologically. The effect of ONO 5046.Na on migration of isolated neutrophils was determined. W/D was not significantly different at 2 h, but was increased at 24 h in the LPS-treated animals. This increase was attenuated in the LPS-treated animals injected with ONO-5046.Na. Analysis of BAL fluid revealed that both at 2 and 24 h after LPS administration the total cell number and neutrophil number, albumin concentration, and elastase-like activity were significantly lower in the LPS treated animals injected with ONO-5046.Na than in those given LPS alone. Histological examination of the lungs of the animals treated with LPS alone showed intra-alveolar haemorrhages and inflammatory cell infiltration 24 h after LPS administration, whereas the lungs of the LPS-treated ONO-5046.Na injected animals were only sparsely infiltrated by inflammatory cells, as indicated by the inflammation score.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7489794 TI - Interstitial pneumonitis-like lesions in guinea-pigs following repeated exposure to toluene diisocyanate. AB - The inhalation of isocyanates, such as toluene diisocyanate (TDI), diphenylmethane diisocyanate (MDI), and hexamethylene diisocyanate (HDI) can induce hypersensitivity pneumonitis (HP) as well as bronchial asthma in humans, but the precise pathological features and their pathogenetic mechanisms have not been elucidated. To provide insight into the pathological features of isocyanate induced hypersensitivity pneumonitis in humans, we repeatedly exposed guinea-pigs to TDI following previous sensitization to TDI and examined the inflammatory response in the pulmonary lesions. Following sensitization with 10% TDI ethyl acetate solution for seven consecutive days, guinea-pigs were exposed to 5% TDI ethyl acetate solution once a week for 4 weeks. As a control, guinea-pigs were exposed to ethyl acetate alone in the same manner. Furthermore, other guinea-pigs received a single exposure to 5 or 20% TDI ethyl acetate solution. The TDI solutions or ethyl acetate were applied to the bilateral nasal mucosa of guinea pigs for 30 s-day-1. Histological examination of lung specimens of guinea-pigs repeatedly exposed to TDI after previous sensitization by TDI inhalation revealed interstitial pneumonitis-like lesions in which mononuclear cells and eosinophils were mainly involved. Lungs of control and nonsensitized guinea-pigs showed insignificant histological changes. We demonstrated that interstitial pneumonitis like lesions, indistinguishable from isocyanate-induced hypersensitivity pneumonitis in humans, can be caused by repeated but not single exposure to TDI in guinea-pigs. PMID- 7489795 TI - Forced oscillation technique (FOT): a new tool for epidemiology of occupational lung diseases? AB - The aim of this study was to evaluate the usefulness of the forced oscillation technique (FOT) in the assessment of occupation-related airway changes. The forced oscillation technique and conventional lung function tests were applied in 80 underground coalface workers, aged 35-48 yrs, with chest roentgenogram films classified 0/1 or 1/0 according to the International Labour Office (ILO) classification (G group), and two control groups matched for age and smoking habits. The first control group, was made up of face-workers having normal chest radiographs, whilst the second comprised underground non-face-workers with normal chest radiographs. Spirometric, plethysmographic and transfer factor of the lungs for carbon monoxide single-breath (TL,CO,sb) indices revealed no significant differences between the three groups. As regards the forced oscillation technique, a higher value of resistance/frequency slope (Pa.L-1.s2) was found in the G group compared with the control groups; 2.11 vs 1.06 in the face-workers, and 1.58 in the underground workers. In all three groups, the forced oscillation technique indices (mean resistance (R), resistance at zero frequency (RO), resistance/frequency slope (S), and resonant frequency (fo)) were found to be higher in subjects having a decreased forced expiratory volume in one second (FEV1) (< or = 90% predicted) or a mildly obstructive pattern of ventilatory function, even though this did not reach statistical significance in each of the groups. These findings together with the feasibility and acceptability of the forced oscillation technique would suggest that it may be a suitable tool for epidemiological studies of occupational respiratory diseases. PMID- 7489796 TI - Exercise-induced airway narrowing in army recruits with a history of childhood asthma. AB - The aim of this study was to determine whether the addition of cold, dry, inspired air to maximum heart rate (MHR) exercise increased the number of subjects demonstrating abnormal airway narrowing, when applied to symptom-free army recruits with a history of asthma in childhood. One hundred recruits with a history of asthma in childhood underwent two exercise challenges to maximum heart rate, breathing either room air or cold dry air in randomized order. The percentage fall index (%FI) was calculated for each challenge, a positive result being > or = 15%. A total of 29 subjects had a positive % fall index, 12 subjects were positive to both challenges, 7 to the room air alone and 10 to the cold air challenge alone. In these subjects (n = 29), the mean (SD) % fall index to the cold air challenge was 22.7 (11.0)% compared with 16.3 (9.5)% for the room air challenge. We conclude that when employed as a screening test for abnormal airway narrowing, the addition of cold, dry, inspired air during exercise does not increase the number of subjects with a positive response, but the magnitude of the response is increased. PMID- 7489797 TI - Preventive therapy for asthma in children; a 9-year experience in eastern Finland. AB - The long-term treatment modalities of bronchial asthma were studied in children from a defined Finnish population from 1985 to 1993, with special reference to changes during the study period. The data on maintenance drugs in children with asthma from five years (1985, 1987, 1989, 1991 and 1993) were retrospectively retrieved from the computerized registers. The reliability of the data for the diagnosis and basic treatment of asthma was checked by one of the authors, who compared the data with the patient cards from the hospital. The number of children with doctor-diagnosed asthma increased continuously during the surveillance period. The proportion of children receiving preventive medication increased concomitantly; this increase was most pronounced between 1987 and 1989. The most common preventive drug was sodium cromoglycate; its use increased from 14% in 1985 to 58% in 1993. The use of inhaled steroids remained stable at 17-19% in all surveillance years. Our treatment policy is in accordance with the international consensus statement published in 1989; however, the change towards preventive medication occurred before its publication. PMID- 7489800 TI - Chronic mucus hypersecretion in COPD and death from pulmonary infection. AB - The association of chronic mucus hypersecretion and mortality is a matter of debate. We wished to determine whether the relationship between chronic mucus hypersecretion and chronic obstructive pulmonary disease (COPD)-related mortality could be explained by proneness to pulmonary infection. We followed 14,223 subjects of both sexes for 10-12 yrs. Cases where COPD was an underlying or contributory cause of death (n = 214) were included, and hospital records were obtained when possible (n = 101). From the presence of increased mucus, purulent mucus, fever, leucocytosis and infiltration on chest radiography, death was classified as either due to pulmonary infection (n = 38), other causes (n = 51), or unclassifiable (n = 12). Of subjects reporting chronic mucus hypersecretion at the initial examination, pulmonary infection was implicated in 54% of deaths, whereas this only occurred in 28% of subjects without chronic mucus hypersecretion. Controlling for covariates, in particular smoking habits, a Cox analysis showed a strong inverse relationship between ventilatory function and COPD-related mortality. Chronic mucus hypersecretion was found to be a significant predictor of COPD-related death with pulmonary infection implicated (relative risk (RR) 3.5) but not of death without pulmonary infection (RR 0.9). We consider that subjects with COPD and chronic mucus hypersecretion are more likely to die from pulmonary infections than subjects without chronic mucus hypersecretion. This may explain the excess mortality in subjects with COPD and chronic mucus hypersecretion found in previous studies. PMID- 7489798 TI - Modulation of the bronchial inflammation in sensitized guinea-pigs by FK506, nedocromil sodium and dexamethasone. AB - Guinea-pigs sensitized by a subcutaneous injection of ovalbumin in Al(OH)3 and boosted 2 weeks later exhibit marked bronchial hyperresponsiveness to various agonists and intense bronchial wall infiltration by CD4+ T-lymphocytes and eosinophils. We have compared the effect of FK506, a novel immunosuppressive agent, on the mucosal infiltration by T-cells and eosinophils with the well established drugs, nedocromil sodium and dexamethasone. Sensitized Hartley guinea pigs were treated subcutaneously for 5 days with FK506 (100 micrograms.kg-1 daily), nedocromil sodium (30 micrograms.kg-1 daily), or dexamethasone (200 micrograms.kg-1 daily). On the day of the experiment, i.e. one week after the booster injection of antigen, the animals were killed, the lungs dissected, frozen and cryostat sections stained by immunohistochemical methods using monoclonal antibodies specific for total T-lymphocytes, CD4+ and CD8+ T-cells. Cyanide-resistant eosinophil peroxidase activity was used to stain the eosinophils. Sections were coded and positive cells enumerated in the lamina propria and adventitia of the bronchi. Sensitized and antigen-stimulated vehicle treated guinea-pigs showed marked infiltration of the bronchial wall by CD4+ T lymphocytes and eosinophils compared with sensitized, non-antigen stimulated animals. As compared to vehicle, FK506 or dexamethasone abolished the T cell/eosinophil invasion in the bronchial wall, whereas nedocromil sodium was ineffective in protecting the lungs from T-lymphocyte or eosinophil infiltration. We conclude that both FK506 and dexamethasone are effective in curtailing bronchial inflammation in allergic guinea-pigs, whereas nedocromil sodium did not resolve the inflammation associated with T-lymphocytes or eosinophils. PMID- 7489799 TI - Influence of endothelial nitric oxide on neurogenic contraction of human pulmonary arteries. AB - The present study was designed to investigate the contribution of the endothelium and that of the L-arginine pathway on the contractile responses of isolated human pulmonary arteries to electrical field stimulation (EFS) and noradrenaline. Isometric tension was measured in artery rings obtained from portions of human lung after thoracic surgery for removal of lung carcinoma (18 patients). Electrical field stimulation (EFS) induced frequency-dependent contractions of isolated human pulmonary arteries which were abolished by tetrodotoxin, guanethidine and prazosin (all at 10(-6) M). The increases in tension were of greater magnitude in arteries denuded of endothelium. NG-nitro-L-arginine methyl ester (L-NAME) (10(-4) M) potentiated the contractile response to EFS in artery rings with endothelium but not in endothelium-denuded arteries. The potentiation induced by L-NAME was completely reversed by L-arginine (10(-4) M) but not by D arginine (10(-4) M). Indomethacin (3 x 10(-6) M) had no significant effect on the contractile response to EFS. Contractile responses to noradrenaline were similar in arteries with and without endothelium. Our results suggest that electrical field stimulation releases endothelium-derived nitric oxide, which inhibits the contractile responses of human pulmonary arteries. Although adrenergic nerves seem to be responsible for the contraction, the transmitter involved in the release of nitric oxide does not appear to be noradrenaline. PMID- 7489801 TI - Functional outcome of patients with chronic obstructive pulmonary disease and exercise hypercapnia. AB - Chronic hypercapnia is associated with a poor prognosis in chronic obstructive pulmonary disease (COPD). Some patients are normocapnic at rest but retain CO2 during exercise. The significance of this abnormality on the course of the disease is unknown. Sixteen stable COPD patients (13 males and 3 females, aged 60 +/- 5 yrs, mean +/- SD) who had previously undergone pulmonary function tests and progressive exercise testing with arterial blood sampling at rest and maximal capacity, entered the study. At first evaluation (E1), subjects were normocapnic at rest (arterial carbon dioxide tension (Pa,CO2): 4.9-5.7 kPa, (37-43 mmHg)) and all presented exercise-induced hypercapnia (end-exercise Pa,CO2 > 5.7 kPa (43 mmHg) with a minimal 0.5 kPa (4 mmHg) increase from resting value). The subjects were re-evaluated 24-54 months later (34 +/- 8 months) (second evaluation (E2)). At E2, forced expiratory volume in one second (FEV1) had decreased from 42 +/- 13 to 38 +/- 15% of predicted values, and mean resting Pa,CO2 had increased from 5.2 +/- 0.3 to 5.7 + 0.4 kPa. Maximal exercise capacity (Wmax) decreased between E1 and E2 from 76 +/- 30 to 56 +/- 22 W. Even if Wmax was lower at E2, end-exercise, Pa,CO2 was higher than at E1 (6.6 +/- 0.8 vs 6.4 +/- 0.5 kPa). At E2, eight subjects presented resting hypercapnia (group H), whilst the others remained normocapnic (Group N). Group H subjects had higher Pa,CO2, at Wmax than Group N and lower Wmax than Group N at E2.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7489802 TI - Metabolic and ventilatory response pattern to arm elevation in patients with COPD and healthy age-matched subjects. AB - Many patients with chronic obstructive pulmonary disease (COPD) experience problems in those activities of daily living which involve so-called unsupported arm elevations (AE). In this study, possible differences in the metabolic and ventilatory response pattern to three types of arm elevation were compared between 13 COPD patients (forced expiratory volume in one second (FEV1) (mean (SD)) 38 (13)% of predicted) and 13 age-matched healthy subjects. Each subject consecutively performed three types of arm elevation for 2 min, with 3 min rest periods in between. Breath-by-breath metabolic and ventilatory parameters and heart rate (HR) were registered during a 3 min baseline period and throughout the measurement period. Adjusted for significantly greater baseline test results in COPD patients, it was found that AE (arm elevation including recovery) tended to be more demanding with respect to metabolic and ventilatory response for patients with COPD than for the age-matched healthy subjects. Arm elevation resulted in an increase in oxygen consumption (V'O2) (COPD 12%; healthy 6%), carbon dioxide elimination V'CO2 (COPD 15%; healthy 10%), minute ventilation V'E (COPD 13%; healthy 7%) and heart rate (COPD 2%; healthy 5%). A pronounced difference was found in the pattern of metabolic and ventilatory response to arm elevation, i.e. there was an earlier but sluggish V'O2 onset in COPD patients, whereas the healthy subjects demonstrated a sudden peak approximately 30 s after arm elevations. Mutual comparison of the three different types of arm elevation demonstrated a comparable response pattern. Knowledge of the specific response to arm elevations in COPD patients seems essential for interpretation of arm elevation tests in upper extremity rehabilitation programmes. PMID- 7489803 TI - Fibrinolytic and inflammatory processes in pleural effusions. AB - This study was designed to evaluate major fibrinolytic parameters in relation to parameters of inflammation associated with different kinds of pleural effusion. Sixty patients with pleural effusion were studied. The underlying aetiology was empyema in 10 cases, tuberculosis in 9, cancer in 31, cardiac failure in 7, and undetermined in 3. Plasminogen, plasminogen activator inhibitor 1 (PAI-1) and 2 (PAI-2), tissue type plasminogen activator (t-PA), urokinase (u-PA) and D-dimers (D-D) were quantified in plasma samples and pleural effusion specimens. These data were then correlated with inflammatory or infectious parameters, i.e. fibrinogen, von Willebrand factor (vWF), erythrocyte sedimentation rate (ESR), protein concentration, and white blood cell count. D-D levels were higher in pleural fluid than in plasma. D-D levels were not correlated with either plasminogen activator or plasminogen activator inhibitor levels, suggesting the presence of other fibrinolytic pathways. PAI levels (PAI activity, PAI-1 antigenicity, PAI-2 antigenicity) and vWF levels were significantly higher in patients with tuberculosis and empyema than in patients with cancer or cardiac failure. Regression analysis between inflammatory and fibrinolytic parameters showed that pleural PAI levels were significantly correlated with pleural neutrophil count, vWF levels, and plasma fibrinogen levels. D-D levels were correlated with blood ESR. No significant difference in pleural t-PA, u-PA and D D levels was observed between aetiologies. The highest pleural t-PA and u-PA values were noted in patients with cancer, especially lymphoma. Plasma t-PA levels were higher inpatients with pleural effusion secondary to congestive heart failure, but this difference did not reach statistical significance.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7489805 TI - Central sleep apnoea, pathogenesis and treatment: an overview and perspective. AB - The prevalence of reported sleep disturbances in a general population is high. Many of the complaints are the result of sleep-related breathing disorders, due mainly to the occurrence of obstructive and central apnoeas. Obstructive sleep apnoea is a fully described and well-recognized entity. Central sleep apnoea (CSA) however, has been poorly studied. There is accumulating evidence that central sleep apnoea should be considered as the end of a spectrum. Instability in the breathing pattern is the main underlying mechanism and is due to the interaction of many factors. Breathing during sleep is dependent on metabolic control and the activity of the respiratory muscles. Decreased chemical drive and/or failing respiratory muscle function are associated with CSA and usually also with ongoing hypoventilation during wakefulness, characterized by chronic daytime hypercapnia. Central respiratory drive can also be inhibited by upper airway reflexes. Mostly, however, CSA occurs as the hallmark of unstable breathing during sleep brought about by an overall increase in loop gain (especially in light sleep stages) and the unmasking of a CO2 threshold. Arousal following central apnoeas acts as an amplification of the instability. Micro electroencephographic (EEG) arousals are often observed as a consequence of CSA. They are responsible for sleep fragmentation and hypersomnolence during the day. The daytime hypersomnolence and complaints of awakenings during sleep in patients with CSA can be striking. CSA can occur in specific pathologies, such as chronic heart failure and (post-traumatic) brain lesions, that are associated with irregular breathing. Treatment strategies are remarkably few in number. Use of nasal ventilation and the inhalation of CO2 are mainly of theoretical interest, since patients do not often tolerate these more invasive therapies. Drug treatment, especially with acetazolamide, is easier to perform. Stimulation of upper airway reflexes, by less invasive methods, seems to be promising for the near future. PMID- 7489804 TI - Effect of oxidant air pollutants on the respiratory system: insights from experimental animal research. AB - In the present paper, we have reviewed experimental animal studies on the effects of the two most important oxidant airborne pollutants, nitrogen dioxide and ozone, on the respiratory system. The toxic effects depend on concentration and length of exposure, and are generally similar for both oxidants, with ozone operative at lower concentrations. High doses of both oxidants cause death due to lung oedema. Exposure to sublethal levels causes functional alterations such as airflow limitation and airway hyperresponsiveness to bronchoconstrictor stimuli. These effects, which are generally reversible, are associated with epithelial injury, oedema and airway and parenchymal infiltration by inflammatory cells. Loss of cilia of airway epithelium and necrosis of type I alveolar epithelial cells are the most prominent consequences at the epithelial level. Inflammation is characterized by early neutrophilic infiltration, followed by an increased number of mononuclear cells, predominantly alveolar macrophages. After long-term exposure, whilst nitrogen dioxide causes predominantly emphysema, ozone produces mainly pulmonary fibrosis. Biochemical effects include lipid peroxidation, increased antioxidant metabolism, and alteration of enzyme activity. Nitrogen dioxide and ozone may also alter the immunological response and reduce the defence against infections, increasing the susceptibility of exposed animals to infections. PMID- 7489806 TI - Anti-tuberculosis medication and the liver: dangers and recommendations in management. AB - In the light of three deaths due to liver failure secondary to anti-tuberculosis therapy at the Royal Free Hospital, we have reviewed the current literature, and asked--How common is liver dysfunction with anti-tuberculosis medications and how might it be prevented? Anti-tuberculosis chemotherapy is associated with abnormalities in liver function tests in 10-25% of patients. Clinical hepatitis develops in about 3%, though estimates vary, and in these patients there is likely to be significant morbidity and mortality. On the basis of reported cases of tuberculosis, 160 patients in England and Wales can be expected to develop drug-induced hepatitis due to anti-tuberculosis therapy each year. There are published guidelines from the British and American Thoracic Societies regarding the choice of drug therapy for tuberculosis. Current recommendations with regard to monitoring liver function, and what to do when these tests become abnormal, vary considerably. We suggest a protocol for using liver function tests to monitor for liver damage, and give recommendations on what action to take when these become abnormal. PMID- 7489808 TI - Optimal assessment and management of chronic obstructive pulmonary disease (COPD). The European Respiratory Society Task Force. PMID- 7489807 TI - Molecular and biological factors in the prognosis of non-small cell lung cancer. AB - For patients with non-small cell lung cancer the tumour/node/metastasis (TNM) staging system and other conventional prognostic factors fail to predict the outcome of treatment and survival accurately. New prognostic factors are urgently needed to improve understanding of the biological behaviour of the different subtypes of non-small cell lung cancer and to recognize patients with a good or poor prognosis. This review will focus on molecular and biological factors published in the English language literature between 1988 and 1994. To be included in this survey, the predictive value of a specific prognostic factor had to be confirmed by multivariate analysis in at least two different studies. Blood group antigen expression, ras oncogenes, microvessel density, and factors reflecting the proliferative state of the tumour may be important determinants of outcome of treatment. The search for new determinants of prognosis has provided insight in the complex tumour biology of non-small cell lung cancer and indicated possible targets for tumour therapy. Several promising prognostic factors have now been recognized. To validate these factors, prospective studies of a large patient population are needed. This ultimately serves the recognition of subsets of patients who may benefit from adjuvant therapy. PMID- 7489809 TI - Occupational asthma due to ceftazidime. AB - A worker employed in the manufacture of the new third generation cephalosporin antibiotic, ceftazidime, developed asthmatic symptoms, and a series of inhalation challenge tests was undertaken to investigate the problem. The inhalation of increasing daily doses of ceftazidime up to 3.2 mg, using a double-blind challenge protocol gave rise to symptoms, late asthmatic reactions, and increases in airway responsiveness to methacholine. A repeat challenge with the 3.2 mg dose additionally gave rise to a clear immediate reaction. Ceftazidime was, thus, shown to be capable of inducing occupational asthma. PMID- 7489810 TI - Normal pregnancy in primary alveolar hypoventilation treated with nocturnal nasal intermittent positive pressure ventilation. AB - Nocturnal nasal intermittent positive pressure ventilation (NIPPV) is increasingly used to treat chronic respiratory failure in a wide variety of conditions (myopathies, Ondine's curse, kyphoscoliosis, etc.) and allows a normal everyday life. Some of these diseases affect women of childbearing age. We report on a young woman suffering from primary alveolar hypoventilation, who presented with limitation of daytime activities, severe oxygen desaturation during sleep, polycythaemia and pulmonary hypertension. These abnormalities completely reversed after a few months of NIPPV applied through a nasal mask. Whilst under ventilatory assistance during sleep, she had an uneventful pregnancy and delivery of a normal baby. We suggest that in selected patients requiring NIPPV, pregnancy can be contemplated with a reasonable level of safety both for the mother and the child, provided that adequate mechanical ventilatory assistance during sleep is maintained throughout pregnancy. PMID- 7489811 TI - Mycobacterium avium-intracellulare pleuritis with massive pleural effusion. AB - Atypical mycobacterial infection is seldom accompanied by pleural involvement. We report a very rare case of Mycobacterium avium-intracellulare pleuritis with massive pleural effusion. The patient was a non-immunocompromised 35-year-old Japanese male with insidious onset of fever, chest pain and anorexia. The pleural effusion gradually resolved with empirical antimycobacterial treatment, leaving considerable pleural adhesion and thickening. PMID- 7489812 TI - Malignant fibrous histiocytoma presenting as an endobronchial polyp of the carina. AB - A 64 year old man was admitted to hospital due to dyspnoea and stridor. A peduncular polyp of the carina was found at bronchoscopy. Histological examination of the tumour after resection with endoscopic electronsurgery revealed malignant fibrous histiocytoma (MFH) of myxoid type. With endoscopic neodymiumyttrium aluminium garnet (Nd-YAG) laser surgery, residual tumour was eliminated. To our knowledge this is the first case with malignant fibrous histiocytoma appearing as a polyp of the carina. PMID- 7489813 TI - A patient with seizures, haemoptysis and dyspnoea. PMID- 7489814 TI - High dose and low dose systematic corticosteroids are equally efficient in acute severe asthma. PMID- 7489815 TI - Predictive factors for death in paediatric acute respiratory failure. PMID- 7489816 TI - Defects of androgen receptor function: from sex reversal to motor neurone disease. AB - The androgen receptor (AR) is a ligand-dependent DNA transcription factor that binds androgens which cause masculinisation of the developing male fetus. Classical abnormalities of receptor function result in the syndrome of androgen resistance, with resultant failure of normal male differentiation. In more recent years, however, mutations in the AR gene have been described in a number of diverse clinical conditions, from male infertility to prostate and breast cancer through to a form of motor neurone disease (Kennedy's disease). This review discusses the various AR gene mutations found in androgen insensitivity syndrome (AIS) and the other conditions described above, and relates how different mutations, or disruption of different functional domains, contributes to the various phenotypes. Mutations that cause complete AIS usually disrupt the DNA or steroid binding ability of the receptor. In partial AIS, mutations generally decrease receptor affinity for ligand, affect thermostability of the protein, or affect the ability of the receptor to activate transcription of responsive genes. Isolated mutations occur in the steroid binding domain of the receptor in prostate cancer, and many cancers have an identical mutation. Similarly, in the two cases of male breast cancer in which AR gene mutations have been described, the mutations in the DNA binding domain of the receptor are alike. In Kennedy's disease a trinucleotide repeat expansion occurs in exon A of the AR gene, which appears to affect ability of the receptor to bind ligand and activate transcription, although the mechanism of neuronal degeneration remains unknown. PMID- 7489817 TI - Differential regulation of AP1 activity by retinoic acid in hormone-dependent and -independent breast cancer cells. AB - We have studied the role of the AP1 transcription factor in the progression of human breast carcinomas. This progression is characterized by a loss of dependence for proliferation on mitogenic hormones, and is also linked to loss of responsiveness to the growth inhibitor retinoic acid (RA). In the hormone dependent breast tumor cell line MCF7 mitogenic stimulation was found to be linked to an enhancement of AP1 transcriptional activity, while growth inhibition by RA was parallelled by decreased AP1 activity. AP1 binding activity to its consensus DNA sequence was rapidly reduced in RA treated cells, in the absence of any noticeable change in expression of AP1 constituents. AP1 overexpression abrogated RA repression in MCF7 cells. In hormone-independent cell lines (BT20, Hs578T, MDA-MB231, MDA-MB468) autonomous proliferation was associated with an increased background AP1 activity. Interestingly, these cells are refractory to growth inhibition by RA, which can only be partly explained by underexpression of RA receptors. In these cells RA did not repress AP1 transactivation unless RA receptors were overexpressed by means of cotransfection with an expression vector. This suggests that the high background levels of AP1 activity in the autonomously growing cells are associated with prevention of RA inhibition of AP1 activity to occur. Therefore, increased AP1 activity may not only play a role in progression of breast tumors towards hormone-insensitivity but may also contribute to the RA resistance of such cells. PMID- 7489820 TI - Modulation by melatonin of protein secretion from melanoma cells: is cAMP involved? AB - The pineal hormone melatonin modulates constitutive protein secretion from melanoma M2R cells. Nanomolar concentrations of melatonin inhibited protein secretion early after plating or at low cell density, but facilitated it late after plating or at high cell density. Inhibition by melatonin of adenylate cyclase is the best known downstream response to melatonin. We have therefore examined the involvement of cAMP in the melatonin-mediated modulation of protein secretion from the melanoma cells. Melatonin slightly but significantly reduced cell cAMP content when effecting inhibition and marginally increased cAMP levels when effecting facilitation of protein secretion. Dibutyryl cAMP abrogated the melatonin-mediated inhibition but not facilitation of protein secretion without affecting basal secretion. Accordingly, forskolin prevented the inhibitory action of melatonin on protein secretion without affecting basal secretion. The selective protein kinase A inhibitor H-89 did not alter the inhibitory effect of melatonin at low cell density and slightly facilitated secretion at high cell density with or without melatonin. Thus, melatonin's effects on protein secretion may not be mediated via cAMP. Nevertheless, changes in cAMP or protein kinase A activity can abrogate, or mask, the melatonin-mediated responses. PMID- 7489819 TI - The complete amino acid sequence of chum salmon stanniocalcin, a calcium regulating hormone in teleosts. AB - Stanniocalcin (STC) is a hypocalcemic hormone secreted from the corpuscles of Stannius of bony fish. Chum salmon STC was isolated in pure form by ion-exchange chromatography and reversed phase high performance liquid chromatography following ethanol-ammonium extraction of the tissues. A cDNA was cloned from cDNAs of the tissue by the PCR method using two primers corresponding to the N- and C-terminal amino acid sequence of the hormone. Sequence analysis of the protein and the cDNA revealed that chum salmon STC is a homodimer, and that the monomer consists of 179 amino acids including 11 half-Cys residues and one N linked glycosylation site, which is 44 residues smaller at the C-terminal region than the sequence deduced from coho salmon STC cDNA. PMID- 7489818 TI - Pituitary PRL secretion induced by tetraethylammonium is inhibited by dopamine through D2 receptors. AB - We have evaluated the inhibitory effect of dopamine on PRL secretion induced by blocking K+ channels. Tumor-derived GH4C1 cells and collagenase-dispersed normal anterior pituitary (AP) cells from young adult male rats were perifused with Krebs-Ringer Hepes medium. In both cell types blocking K+ channels with tetraethylammonium (TEA) induced PRL secretion but did not stimulate cyclic AMP generation. Blocking Na+ channels with 1 microM tetrodotoxin had no effect on basal or TEA-induced PRL secretion. Dopamine inhibited the TEA-induced rise in [Ca2+]i in GH4C1 cells expressing dopamine D2 short receptors. In normal AP cells, 1-100 nM dopamine blocked PRL secretion induced by 20 mM TEA in a log linear concentration-dependent fashion, with a plateau at > 100 nM dopamine (IC50 30 nM). The D2 dopaminergic receptor agonist, quinpirole, at 100 nM completely blocked PRL secretion induced by 20 mM TEA. The D2 dopaminergic receptor antagonist, sulpiride, at 10 microM reversed the inhibitory effect of 10 microM dopamine on PRL secretion induced by 20 mM TEA. Pretreatment of cells with 100 ng/ml pertussis toxin (PTX) for 24 h prevented 100 nM dopamine inhibition of PRL secretion induced by 20 mM TEA. The data indicate that in both normal lactotroph cells and in tumor-derived cells expressing D2 receptors, PRL secretion stimulated by blocking K+ channels is inhibited by dopamine binding to D2 receptors on the plasma membrane. This inhibition involves interaction with PTX sensitive Gi protein. PMID- 7489822 TI - Sex differences in cytochromes oxidase and P-45011 beta in the rat adrenal cortex. AB - A comparative study of cytochrome c oxidase (COX) activity and expression as well as cytochrome P-45011 beta expression has been carried out on the adrenal cortex of male and female rats. COX has also been examined in rat liver. In addition, the effect of testosterone replacement in orchiectomized male rats on adrenal COX has also been investigated. Adult male rats had higher COX activity in adrenal (255%) and liver (144%) mitochondria compared to adult female rats. Male rat adrenals and liver also had increased levels of COX II, a mitochondria-encoded COX subunit, and of COX IV, a nucleus-encoded COX subunit, as measured by Western analysis. In contrast, cytochrome P-45011 beta levels were lower (48%) in adrenal mitochondria from male rats than those of female rats. There was no significant sex difference in the level COX II and COX IV mRNAs in adrenal or liver, whereas the cytochrome P-45011 beta mRNA was 4-fold higher in female adrenals than in males. In male rats, orchiectomy caused a 23% decrease and testosterone replacement a 66% increase in adrenal COX activity. There were no corresponding changes in the levels of mRNAs encoding for COX subunits, suggesting post transcriptional effects of testosterone on COX. These results are consistent with a regulatory role of testosterone on the expression of components of the respiratory and steroidogenic electron transport chains. PMID- 7489821 TI - Phospholipase A2 expression in human and rodent insulin-secreting cells. AB - The expression of different isoforms of phospholipase A2 in human and rat islets of Langerhans and in the clonal B-cell lines, HIT-T15 and RINm5F has been investigated, using polyclonal antisera specific to human cytosolic (cPLA2) or human Groups I and II secretory (sPLA2) isoforms. Abundant levels of a 100-kDa protein corresponding to cPLA2 were detected in cytosolic extracts of human islets. A 100-kDa cPLA2 was not detectable in rat islets, RINm5F or HIT-T15 cells using an anti-cPLA2 serum raised against cloned human cPLA2 cDNA, despite the antiserum being cross-reactive with cPLA2 from rat kidney. Human and rat islets were found to express a 21-kDa protein immunoreactive with Group I sPLA2 antiserum. Group II sPLA2 was not detected in human or rat islets. RIN cells did not express detectable levels of either Group I or Group II sPLA2, but HIT cells expressed variable quantities of Group II sPLA2. These differences in PLA2 expression suggest that caution should be exercised when extrapolating conclusions about lipid-derived signalling molecules from insulin-secreting cell lines to normal islets of Langerhans. PMID- 7489824 TI - Heterogeneous response of porcine gonadotrope subpopulations to gonadotropin releasing hormone (GnRH) during postnatal development. AB - In previous papers, we showed the porcine gonadotrope population to be composed of three GtH subpopulations that can be separated by density using a continuous Percoll density gradient. We also demonstrated that these subpopulations exhibited different hormonal storage patterns and morphological features during porcine postnatal development at three representative ages: neonates (30 days), prepubers (5-6 months) and matures (16-18 months). In this work, we investigated whether these morphologically heterogeneous subpopulations are also functionally different. Thus, the effect of the hypothalamic gonadotropic hormone-releasing factor (GnRH) on these subpopulations was assessed in order to ascertain whether a mutual relationship between the reported morphological features, hormonal storage patterns and physiological response to the stimulation can be established. For this purpose, gonadotropin secretion was measured by cell immunoblot assay and hormonal content by scanning cytophotometry. Low-density gonadotropes (1.049 g/cm3), present in the three age groups studied, were mainly composed of bihormonal LH/FSH cells in neonates and monohormonal LH cells in prepubers and matures. GnRH stimulation was found to increase both LH and FSH secretion, as well as the intracellular content. These results indicate that GnRH can stimulate both the synthesis and release of both gonadotropins in this subpopulation. Middle-density gonadotropes (1.062 g/cm3), present in prepubers and matures only, were composed of bihormonal cells. GnRH stimulated the secretion of LH and FSH in prepubers and matures, but decreased hormonal contents except that of LH in prepubers. However, GnRH stimulation increased the proportion of immunoreactive gonadotropes (particularly monohormonal cells). Finally, high-density cells (1.087 g/cm3), present in neonates and prepubers only, were mostly composed of bihormonal LH/FSH gonadotropes, and exhibited low (neonates) or no response (prepubers) in terms of LH release and content when treated with GnRH. In conclusion, these results indicate that porcine gonadotrope subpopulations are morphologically and physiologically heterogeneous. The heterogeneity remained through porcine postnatal development, thus suggesting that all the subpopulations are physiologically relevant. However, the different hormonal storage patterns between subsets of the same density suggest age-related differences within each subpopulation due, at least in part, to the different physiological condition of the animals during development. PMID- 7489823 TI - Growth hormone-releasing factor stimulates meiotic maturation in follicle- and cumulus-enclosed rat oocyte. AB - This study was designed in order to assess the possible role of growth hormone releasing factor (GRF) on oocyte maturation. This effect was analyzed in follicle enclosed, cumulus-enclosed and denuded oocytes obtained from immature pregnant mare's serum gonadotropin-treated rats. The addition of GRF to the cultures significantly accelerated maturation in follicle- and cumulus-enclosed oocytes while no effect was seen on denuded oocytes. Also, the neuropeptide was able to induce maturation in follicle-enclosed oocytes obtained from immature untreated rats. The GRF action was probably not mediated by the vasoactive intestinal peptide (VIP) receptors since the two hormones had different effects on oocyte maturation and on cAMP production by granulosa cells. In addition the disappearance of the GRF effect observed in the presence of antibodies anti-GH suggested that GRF required the intermediacy of GH to accomplish its effect on oocyte maturation. Finally, GRF did not affect meiotic maturation when dbcAMP was added to the cultures. Our results demonstrate the ability of GRF to accelerate maturation in oocytes from both primed and unprimed rats. Since the presence and the involvement of GRF at the ovarian levels is now well established, the present data strongly suggest an important potential role of GRF in the ovarian physiology. PMID- 7489825 TI - Role of a hydrophobic pocket of the human Y1 neuropeptide Y receptor in ligand binding. AB - We are investigating the nature of the chemical interactions between the neuropeptide Y (NPY) and its cell surface receptor (Y1). A previous study involving site-directed mutagenesis and computer-aided modelling (Walker et al., 1994) suggested that the C-terminal Tyr36 of NPY, known to be a key residue for receptor binding, might dock at a pocket formed by hydrophobic amino acids of transmembrane domains (TM) 1, 2, 6 and 7 of the Y1 receptor. To investigate which residues were required for ligand binding, we mutated the sequences encoding F41, L43, F96, Y100, F286 and H298 of the human Y1 receptor. The mutant cDNAs were transiently expressed in Hela cells and the ability of the encoded proteins to bind NPY was evaluated. Replacing F41, L43 or F96 with alanines had no effect on NPY binding. On the contrary, Y100, F286 and H298 appeared to be residues critical for ligand binding. In particular, the removal of the hydroxyl group of Y100 (Tyr100-->Phe100 mutation) yielded a protein devoid of affinity for the ligand. The level of expression and the presence on the cell surface of mutants lacking NPY binding activity was assessed by immunological techniques. In addition, we tested the ability of synthetic analogues of neuropeptide Y with substitutions at position 36 to bind to the Y1 receptor. To get spatial insight into the relative positions of the above mentioned residues we constructed a molecular model of the interaction between NPY:Y36 and the elements of the hydrophobic pocket surrounding this residue. PMID- 7489826 TI - Biological activity of phosphorylated and dephosphorylated bovine prolactin. AB - Phosphorylation is a mechanism by which cells regulate structure and function of proteins. We have previously demonstrated in vivo synthesis and secretion of phosphorylated bovine prolactin (bPRL) from the pituitary, and have isolated and partially characterized the phosphorylated bPRL. In order to investigate the structure/function role of phosphorylation on the biological activity of bPRL, we compared the activities of nonphosphorylated and phosphorylated bPRL isolated from pituitaries, with bPRL provided by the NIDDK (NIDDK-bPRL) to stimulate Nb2 cell proliferation. Nonphosphorylated bPRL has activity similar to, although slightly lower than that of NIDDK bPRL (ED50 = 7.03 pM and 22.8 pM, respectively). The activity of phosphorylated bPRL is significantly reduced (ED50 = 1066 pM). Using Nb2 lymphoma cell homogenate, NIDDK and nonphosphorylated bPRLs are equally effective in competitive receptor binding assays (Kd = 0.252 and 0.269 nM, respectively). Phosphorylated bPRL does not compete for the PRL receptor at concentrations up to 65 nM. Following enzymatic removal of the phosphate group using alkaline phosphatase, there is an increase in the biological activity of phosphorylated bPRL (ED50 = 73.3 pM) while the activity of nonphosphorylated BPRL remained unchanged following enzyme treatment (21.4 pM). We conclude that (1) structural changes induced by phosphorylation of bPRL are responsible for loss of bioactivity, (2) dephosphorylation of phosphorylated bPRL restores biological activity, and (3) the reduction in biological activity of phosphorylated bPRL is mediated by a decrease in receptor binding. PMID- 7489828 TI - A novel human cDNA highly homologous to the fish hormone stanniocalcin. AB - Stanniocalcin is a glycoprotein hormone previously considered present only in bony fish where it is secreted by the corpuscles of Stannius, endocrine organs involved in Ca2+ homeostasis. In fish, stanniocalcin was thought to be an adaptation for Ca2+ regulation in aquatic environments, and its effects include inhibition of gill Ca2+ transport. We have obtained a human cDNA clone coding for a protein highly homologous to fish stanniocalcin. The mRNA is expressed in many human tissues, with the highest levels in ovary, prostate and thyroid. In vitro human cell culture studies show that the mRNA is positively regulated by extracellular Ca2+ in the medium. We conclude that a human protein similar to the fish hormone is expressed in multiple tissues rather than by a specialized endocrine organ. PMID- 7489827 TI - Keratin expression and steroidogenesis in rat granulosa cells, transformed with the Kirsten-ras and SV40 oncogenes singly and in combination. AB - The keratins are a component of the cytoskeleton that is present in fetal and neonatal rat granulosa cells (ROG), but disappears as the cells undergo postnatal steroidogenic differentiation. Steroidogenesis is initiated in the fetus as a low level constitutive function which is cAMP responsive, but becomes responsive to gonadotrophic hormones only after birth. ROG from PMSG-primed immature rats, like mature ROG, are keratin negative, highly steroidogenic and gonadotrophin responsive, but rapidly lose their steroidogenic capacity in culture. In such cultured cells, transformation with the Kirsten-ras oncogene (v-Ki-ras) maintains low levels of constitutive steroidogenesis and responsiveness to cAMP, and induces the expression of keratin. To determine whether similar changes would occur in cells expressing both the SV40 and v-Ki-ras, cultured ROG were transformed with SV40 early genes, with Kirsten murine sarcoma virus (KiMSV), or with both agents concurrently. Keratin was demonstrated by fluorescence microscopy and Western blots, and progesterone production by RIA. ROG transformed with SV40 alone became immortalized but secreted little steroid and lacked keratin. In contrast, three cell lines, co-transformed with SV40 plus KiMSV, acquired keratin as well as the capacity to secrete progesterone in response to cAMP, closely resembling cells transformed with Ki-ras alone. KiMSV-transformed muscle fascia fibroblasts lacked both steroidogenic potential and keratin. The results show that the complex, v-Ki-ras-induced changes in steroidogenesis and keratin expression are reproducible and tissue specific. The phenotypic resemblance between singly and doubly transformed ROG indicates that the v-Ki-ras oncogene does not act by overcoming SV40-mediated inhibition of differentiation.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7489829 TI - Constitutively active Gq-alpha stimulates prolactin promoter activity via a pathway involving Raf activity. AB - We have investigated the ability of a constitutively active Gq-alpha mutant, Q209L-alpha q, to regulate target gene expression. Transient expression in GH3 pituitary cells of a rat proximal prolactin promoter-chloramphenicol acetyltransferase construct (-187)PRL-CAT, was stimulated by co-expression of Q209L alpha q, but not by wild-type alpha q. Q209L-alpha q stimulated expression of constructs driven by promoters for either rat prolactin or growth hormone, but not of a control construct driven by the thymidine kinase promoter. Thus, transcriptional effects of alpha q are specific both for the activated state of this G-alpha subunit and the promoter examined. Since both the prolactin and growth hormone promoters are activated by the pituitary cell-specific transcription factor Pit-1, we examined whether a Pit-1 binding site could direct a response to Q209L-alpha q. Two copies of prolactin promoter Pit-1 binding site 1P conferred upon a heterologous metallothionein promoter a response to Q209L alpha q, implying an involvement of this site in the transcriptional action of Q209L-alpha q on the prolactin promoter. The phorbol ester activator of protein kinase C, 12-O-tetradecanoylphorbol-13-acetate, stimulated (-187)PRL-CAT activity, but opposed the action of Q209L-alpha q on activity of this PRL-CAT construct. Q209L-alpha q stimulation of (-187)PRL-CAT activity was inhibited by co-expression of a dominant negative Raf mutant, Raf-C4, but not by a point mutant of Raf-C4 with reduced inhibitory properties. These results imply that activated alpha q subunits can stimulate prolactin promoter activity via a pathway that involves a Pit-1 DNA binding site(s), is opposed by protein kinase C, and is mediated by a pathway in which Raf-1 kinase plays a role. PMID- 7489830 TI - Activation of MAP kinase by the LHRH receptor through a dual mechanism involving protein kinase C and a pertussis toxin-sensitive G protein. AB - The LHRH receptor in alpha T3-1 gonadotrope cells was shown to bring about a marked and sustained activation of MAP kinase. This response was prevented by protein kinase C inhibition or down-regulation and could be partially mimicked by phorbol ester. Additional evidence for inhibition of this response by pertussis toxin and partial mimicry by mastoparan (in a pertussis toxin-sensitive manner) provides the first evidence for Gi/Go-mediated signal transduction by the LHRH receptor. This dual mechanism of MAP kinase activation appears to be exceptional amongst the G protein-linked receptors that have been investigated. PMID- 7489831 TI - The NSY mouse: a new animal model of spontaneous NIDDM with moderate obesity. AB - The NSY (Nagoya-Shibata-Yasuda) mouse was established as an inbred strain of mouse with spontaneous development of diabetes mellitus, by selective breeding for glucose intolerance from outbred Jcl:ICR mice. NSY mice spontaneously develop diabetes mellitus in an age-dependent manner. The cumulative incidence of diabetes is 98% in males and 31% in females at 48 weeks of age. Neither severe obesity nor extreme hyperinsulinaemia is observed at any age in these mice. Glucose-stimulated insulin secretion was markedly impaired in NSY mice after 24 weeks of age. In contrast, fasting plasma insulin level was higher in male NSY mice than that in male C3H/He mice (545 +/- 73 vs 350 +/- 40 pmol/l, p < 0.05, at 36 weeks of age). Pancreatic insulin content was higher in male NSY mice than that in male C3H/He mice (76 +/- 8 vs 52 +/- 5 ng/mg wet weight, p < 0.05, at 36 weeks of age). Morphologically, no abnormal findings, such as hypertrophy or inflammatory changes in the pancreatic islets, were observed in NSY mice at any age. These data suggest that functional changes of insulin secretion in response to glucose from pancreatic beta cells may contribute to the development of non insulin-dependent diabetes mellitus (NIDDM) in the NSY mouse. Although insulin sensitivity was not measured, fasting hyperinsulinaemia in NSY mice suggests that insulin resistance may also contribute to the pathogenesis of NIDDM. Since these findings are similar to the pathophysiologic features of human NIDDM patients, the NSY mouse is considered to be useful for investigating the pathogenesis and genetic predisposition to NIDDM. PMID- 7489832 TI - The ACE-inhibitor captopril improves myocardial perfusion in spontaneously diabetic (BB) rats. AB - The aim of this study was to examine the influence of inhibition on angiotensin converting enzyme (ACE) of myocardial function and perfusion of the rat impaired by diabetes. Spontaneously diabetic rats were treated with the ACE-inhibitor captopril for 4 months. Cardiac performance was analysed in the isolated heart perfused at constant volume. Epicardial perfusion was determined by measuring changes in epicardial fluorescence after injection of a bolus of fluoresceinisothiocyanate-dextrane (3 kDa) as described previously. As compared to untreated diabetic controls, captopril prevented the increase of end diastolic pressure, coronary perfusion pressure and vascular resistance. The intravascular volume was enlarged and the epicardial perfusion rate increased in hearts of diabetic rats treated with captopril as compared to diabetic controls. Treatment of diabetic rats with the ACE-inhibitor captopril (1) increases the number of perfused capillaries, and (2) can partly prevent the development of cardiac dysfunction in diabetes. Together with morphological data demonstrating an inhibition of interstitial and perivascular fibrosis in hearts of diabetic rats treated with captopril, our data suggest that ACE-inhibition is cardioprotective in diabetes. These observations are also compatible with the assumption that an accelerated generation of angiotensin II may be involved in the pathophysiological chain of events leading to diabetic cardiopathy. PMID- 7489833 TI - In vivo effects of glucosamine on insulin secretion and insulin sensitivity in the rat: possible relevance to the maladaptive responses to chronic hyperglycaemia. AB - We tested the hypothesis that glucosamine, a putative activator of glucose toxicity in vitro through acceleration of the hexosamine pathway, may determine in vivo the two key features of glucose toxicity in diabetes, namely, peripheral insulin resistance and decreased insulin secretion. Two groups of awake rats were studied either with intraarterial administration of glucosamine (5 mumol.kg-1.min 1) or saline. Insulin secretion was determined after arginine, glucose (hyperglycaemic clamp), and arginine/glucose infusions, while insulin-mediated glucose metabolism was assessed by the euglycaemic hyperinsulinaemic clamp in combination with [3-3H]-glucose infusion. Glucosamine had no effects on arginine induced insulin secretion both at euglycaemia and hyperglycaemia, but significantly (40-50%) impaired glucose-induced insulin secretion (both first and second phases). During euglycaemic hyperinsulinaemic clamp studies, glucosamine decreased glucose uptake by approximately 30%, affecting glycolysis (estimated from 3H2O rate of appearance) and muscle glycogen synthesis (calculated from accumulation of [3H]-glucosyl units in muscle glycogen) to a similar extent. Muscle glucose 6-phosphate concentration was markedly reduced in the glucosamine infused rats, suggesting an impairment in glucose transport/phosphorylation. Therefore, an increase in hexosamine metabolism in vivo: 1) inhibits glucose induced insulin secretion, and 2) reduces insulin stimulation of both glycolysis and glycogen synthesis, thereby mimicking in normal rats the major alterations due to glucose toxicity in diabetes. PMID- 7489835 TI - Prevention of glomerular hyperfiltration in rats with streptozotocin-induced diabetes by an atrial natriuretic peptide receptor antagonist. AB - The contribution of atrial natriuretic peptide (ANP) to the development of glomerular hyperfiltration in diabetes was investigated by examining the effects of HS-142-1, a non-peptide antagonist of biological receptors for ANP, on glomerular filtration rate (GFR) and renal plasma flow (RPF) in rats with streptozotocin-induced diabetes. Three to four weeks after streptozotocin injection, the plasma concentration of ANP, urinary cyclic GMP excretion rate, GFR, and RPF were significantly higher in diabetic rats than in control rats. The increase in GFR and RPF in diabetic rats was significantly reduced, in a dose dependent manner, by a single intravenous injection of HS-142-1; the maximal effect was apparent at a dose of 10 mg per kg of body weight. Continuous subcutaneous administration of HS-142-1 with an osmotic minipump for 3 to 4 weeks, beginning 2 days after streptozotocin injection, prevented the increases in urinary cyclic GMP excretion rate, GFR, and RPF observed in untreated diabetic rats. These results highlight the importance of ANP in the development of diabetic glomerular hyperfiltration and indicate that this condition can be prevented by continuous inhibition of the action of ANP. PMID- 7489834 TI - Tamm-Horsfall glycoprotein in streptozotocin diabetic rats: a study of kidney in situ hybridization, immunohistochemistry, and urinary excretion. AB - Tamm-Horsfall glycoprotein, present only in the kidney thick ascending limb of Henle's loop, was studied here in streptozotocin diabetic rats. Tamm-Horsfall glycoprotein mRNA in situ hybridization was performed on snap-frozen left kidneys; the right kidneys were perfusion-fixed with 4% paraformaldehyde and embedded either in paraffin, for Tamm-Horsfall glycoprotein immunohistochemistry, or in Epon for stereologic measurements. The length of the thick ascending limb of Henle's loop and the amount of glycogen were measured and the ultrastructure of the cells was evaluated. Urinary excretion of Tamm-Horsfall glycoprotein, calcium, magnesium and albumin was measured. After 10 and 50 days' duration of diabetes, kidney weight increased 20 and 41%, respectively and the length of the thick ascending limb of Henle's loop increased 28 and 56%, respectively, compared with controls. Substantial glycogen accumulations were present in the thick ascending limb of Henle's loop, and electron microscopy revealed a significant decrease in organelles and basolateral membranes. After 10 and 50 days' duration of diabetes, in situ hybridization of Tamm-Horsfall glycoprotein mRNA revealed a fourfold decrease, and the immunostaining for Tamm-Horsfall glycoprotein showed a threefold decrease as measured by densitometry. However, urinary Tamm-Horsfall glycoprotein excretion rate was increased fivefold and urinary concentration about twofold. Urinary calcium excretion increased three-fold and magnesium twofold, but urinary albumin excretion was not significantly increased. The increased amount of Tamm-Horsfall glycoprotein, calcium and magnesium in the urine in diabetes occurs here concomitant with severe cellular damage in the thick ascending limb of Henle's loop. PMID- 7489836 TI - Rapid deposition of amyloid in human islets transplanted into nude mice. AB - Human islets of Langerhans were transplanted to the subcapsular space of the kidneys of nude mice which were either normoglycaemic or made diabetic with alloxan. After 2 weeks, the transplants were processed for light and electron microscopical analyses. In all transplants, islet amyloid polypeptide (IAPP) positive cells were found with highest frequency in normoglycaemic animals. IAPP positive amyloid was seen in 16 out of 22 transplants (73%), either by polarisation microscopy after Congo red staining or by immune electron microscopy. At variance with previous findings of amyloid deposits exclusively in the extracellular space of islets of non-insulin-dependent diabetic patients, the grafted islets contained intracellular amyloid deposits as well. There was no clear difference in occurrence of amyloid between diabetic and non-diabetic animals. The present study indicates that human islets transplanted into nude mice very soon present IAPP-positive amyloid deposits. This technique may provide a valuable model for studies of the pathogenesis of islet amyloid and its impact on islet cell function. PMID- 7489837 TI - Incidence of IDDM during 1988-1992 in Zagreb, Croatia. AB - The objective of this study was to determine the incidence of insulin-dependent diabetes mellitus (IDDM) in the population of Zagreb, Croatia, during 1988-1992. A centralized diabetes registry was the primary source of data, while secondary sources were used to assess ascertainment. A total of 282 new cases of IDDM were diagnosed in the study period, the primary and secondary sources identifying annually 93-100% of the cases. The annual incidence rate ranged from 5.6 per 100,000 to 6.6 per 100,000. Early fatality in persons older than 50 years was the major cause of underascertainment. The incidence peaked in the 10-14 years age group (12.4 per 100,000), and remained stable after age 24 years. Males had a significantly higher incidence in the 5-9 and 24-44 years age groups. In the 45 54 years age group, females had a significantly higher incidence. No seasonality was observed. Despite the war conditions in Croatia, the low overall IDDM incidence rates did not change significantly during the study period. PMID- 7489840 TI - Visual evoked potentials in NIDDM: a longitudinal study. AB - In order to assess the possible progression of neurological abnormalities over time and the value of visual evoked potential alterations in predicting stability and severity of diabetes-related optic pathway disease, a longitudinal study in non-insulin-dependent diabetic patients was performed. Neurological examination, visual evoked potentials with pattern reversal, motor and sensory nerve conduction velocities and metabolic control were studied in 18 non-insulin dependent diabetic patients and in 35 normal control subjects at baseline and again after 4.6 +/- 0.8 years (range 4-6). At the first recording the peak P100 wave latencies were significantly delayed in the diabetic patients compared with the control subjects; signs of peripheral neuropathy were detected in five patients, clinical in three and in two there was only neurophysiological alteration without clinical signs. The second recording revealed no significant alterations of P100 latencies in patients compared with baseline, but the number with clinical signs and/or neurophysiological alterations with no clinical signs of peripheral neurological disease was increased to seven. In conclusion, we observed that visual evoked potential alterations were stable over time whereas peripheral neurological disease progressed and correlated positively with metabolic control. PMID- 7489838 TI - Methodological aspects, dose-response characteristics and causes of interindividual variation in insulin stimulation of limb blood flow in normal subjects. AB - To resolve some of the controversy regarding insulin regulation of blood flow, we performed in 20 normal subjects a) a reproducibility study of plethysmographic, Doppler ultrasound and laser Doppler blood flow measurements (n = 7), b) a sequential insulin dose-response study with measurement of forearm (plethysmography), leg (Doppler ultrasound) and skin (laser Doppler) blood flow (n = 12), and c) a sequential insulin dose-response study with comparison of forearm (plethysmography) and calf (plethysmography) blood flow (n = 8). We also searched for factors which might explain the interindividual variation in the blood flow response to insulin. During sequential insulin infusions (2 h each, 61 +/- 2, 139 +/- 6, 462 +/- 15 mU/l), forearm blood flow increased by 17 +/- 6, 50 +/- 14 and 113 +/- 17% (p < 0.05 or less between steps), respectively. The increase at the 61 +/- 2 mU/l insulin concentration barely exceeded methodological variation (13 +/- 2%). In contrast to the continuous increase in blood flow, the glucose arterio venous difference reached its maximum (1.7 +/- 0.2 mmol/l) at the lowest 61 +/- 2 mU/l insulin concentration and remained constant thereafter. Forearm and calf blood flow responses to insulin were virtually identical when determined with plethysmography. In contrast, only a 27% increase was detected in femoral flow index as determined by Doppler ultrasound. Forearm blood flow (per forearm volume) was highly correlated with the relative forearm muscle content (mean 59 +/- 5%, range 24-81%) both basally (r = 0.86, p < 0.001, n = 12) and at all insulin concentrations (r = 0.85-0.92, p < 0.001) indicating that the percent of forearm that is muscle explains 70-85% of interindividual variation in blood flow. In conclusion 1) physiological insulin concentrations stimulate glucose uptake mainly by increasing glucose extraction while supraphysiological insulin concentrations increase forearm glucose uptake predominantly via increases in blood flow. 2) The dose-response characteristics of insulin stimulation of forearm and calf blood flow are similar when determined with strain-gauge plethysmography. 3) Relative forearm muscle content is a key factor in determining both basal forearm blood flow and the interindividual variation in its response to insulin in normal subjects. PMID- 7489841 TI - Immunogenicity of intraperitoneal insulin infusion using programmable implantable devices. AB - Intraperitoneal insulin infusion using implantable devices in insulin-dependent diabetic (IDDM) patients is promising since it improves diabetic control and decreases frequency of hypoglycaemia. However, preliminary data show a striking increase in plasma levels of anti-insulin antibodies with this therapy. In order to more precisely evaluate the immunogenicity and its consequences, anti-insulin antibody levels in 62 IDDM patients were assessed every 3 months during a 2-year period following pump implantation. At the same time, diabetes control was evaluated with HbA1c, mean blood glucose levels, standard deviation of the daily blood glucose levels and the frequency of low blood glucose (< 3.58 mmol/l). Factors involved in antibody formation such as age, gender, HLA typing, and complement C4 alleles were also studied. After implantation, anti-insulin antibody levels increased significantly from 3.14% (range 0-26%) to 8.34% (0-49%) after 1 year and remained elevated. Patients were divided into two groups: responders able to show at least one antiinsulin antibody titre higher than 15% and non-responders whose titres were always lower than 6%. None of the factors studied was shown to statistically influence the anti-insulin antibody titres. Non-responders had significantly better metabolic results than the responders. Severe hypoglycaemic episodes decreased dramatically in both groups. Insulin requirements were comparable at time 0 and decreased initially in both groups. They remained low for the non-responders but returned to pre-implantation values for responders. Intraperitoneal insulin infusion led to a high immunogenetic response towards insulin in about half of the patients, leading to only moderately deleterious effects on metabolic control.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7489842 TI - Asymptomatic hyperglycaemia is associated with increased intimal plus medial thickness of the carotid artery. AB - Atherosclerotic changes have not been demonstrated directly in asymptomatic hyperglycaemic non-diabetic subjects, although high mortality due to coronary heart disease has been reported. We measured arterial wall thickness non invasively, in order to directly demonstrate atherosclerosis of the carotid arteries of hyperglycaemic non-diabetic subjects and to evaluate its risk factors. The thicknesses of the intimal plus medial complex (IMT) of the carotid arteries of 112 asymptomatic hyperglycaemic non-diabetic subjects (aged 22-81, 95 males and 17 females) were compared with those of 55 healthy male subjects and 211 non-insulin-dependent NIDDM male diabetic patients. The subjects were subgrouped into impaired glucose-tolerant (IGT) subjects who had a 2-h glycaemic level of more than 7.8 mmol/l, and non-IGT subjects whose 2-h glycaemic levels were within 6.7-7.7 mmol/l. Non-IGT and IGT subjects showed significantly greater IMTs than age-matched healthy males and showed no significant differences compared to age-matched NIDDM patients. Multivariate analysis demonstrated that the risk factors for IMT of non-IGT and IGT subjects were age and systolic blood pressure. According to data on the accumulation of atherogenic risks (hypertension, dyslipidaemia, and smoking), IMT increased linearly in non-IGT and IGT subjects. However, non-IGT and IGT subjects without hyperlipidaemia, hypertension, or smoking risk still had significantly greater IMT than age matched normal males (1.019 +/- 0.063 vs 0.770 +/- 0.111 mm, p < 0.05).(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7489839 TI - Insulin resistance and hyperinsulinaemia in mild to moderate progressive chronic renal failure and its association with aerobic work capacity. AB - Tissue sensitivity to insulin and aerobic work capacity was measured in patients with mild to moderate progressive chronic renal failure. Twenty-nine non-diabetic patients with a glomerular filtration rate of 25 ml.min-1.1.73 m-2 (11-43) (median, range) and 15 sex, age, and body mass index matched control subjects with normal renal function were studied. Fasting blood glucose was comparable and in the non-diabetic range in the two groups as was the oral glucose tolerance test. Patients demonstrated hyperinsulinaemia both during fasting (p < 0.01) and during the test (p < 0.02). The tissue sensitivity to insulin, expressed by the amount of glucose infused during the last 60 min of a 120-min hyperinsulinaemia euglycaemic clamp (M-value) and the M/I ratio, was significantly lower in the patients than in the control subjects (M-value 404 +/- 118 vs 494 +/- 85 mg glucose/kg body weight, p < 0.02) (M/I ratio 1.77 +/- 0.71 vs 2.57 +/- 0.70 (mg/(kgBW.min) per pmol/l.100, p < 0.001). The maximal aerobic work capacity was significantly lower in the patients than in the control subjects (24 +/- 8 vs 32 +/- 11 ml O2/(kg body weight.min), p < 0.02) and positively correlated to the M value and the M/I ratio in both groups. In conclusion, not only patients with end stage chronic renal failure but also those with mild to moderate progressive chronic renal failure are insulin resistant and hyperinsulinaemic. The tissue sensitivity to insulin is correlated to the maximal aerobic work capacity suggesting that these patients might benefit from physical training programmes. PMID- 7489843 TI - Long-term comparison of human insulin analogue B10Asp and soluble human insulin in IDDM patients on a basal/bolus insulin regimen. AB - Recombinant DNA technology allows the production of insulin analogues with faster absorption rates from subcutaneous tissue as compared to soluble human insulin. The human insulin analogue B10Asp (mono/dimeric) is absorbed twice as fast as soluble human insulin (hexameric). A double blind, randomised crossover study with a 1-month run-in period and two 2-month treatment periods was performed in 21 male insulin-dependent diabetic (IDDM) patients aged 18-40 years in order to compare the metabolic control obtained with equimolar doses of the analogue B10Asp vs soluble human insulin (Actrapid) given as mealtime insulin and intermediate acting isophane insulin (Protaphane) at bedtime. At the end of each 2-month study period, the patients were admitted to the metabolic ward. We found significantly higher plasma insulin/analogue levels after breakfast, lunch and dinner with B10Asp as compared to Actrapid (p < 0.05). The plasma insulin/analogue levels were significantly lower before lunch and dinner with B10Asp as compared to Actrapid (p < 0.05). Also, the plasma insulin/analogue level tended to be lower at bedtime when comparing B10Asp to Actrapid. The 24-h blood glucose profiles showed identical fasting blood glucose, significantly lower blood glucose after breakfast with the analogue (p < 0.05), no differences in blood glucose after lunch and dinner but a significantly higher blood glucose at midnight using the analogue (p < 0.05). The overall 24-h mean blood glucose concentrations, the daily insulin dose, HbA1c, diet, home blood glucose monitoring and frequency of hypoglycaemia were almost identical in the two treatment periods.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7489844 TI - Blood pressure, retinopathy and urinary albumin excretion in IDDM: the EURODIAB IDDM Complications Study. AB - Several studies have shown an association between blood pressure and nephropathy, but few have been large enough to examine whether, or how, this relation is influenced by retinopathy. We have therefore examined the independent relations of blood pressure to urinary albumin excretion and retinopathy in a cross sectional observational study of over 3000 insulin-dependent diabetic patients (the EURODIAB IDDM Complications Study). The relation of blood pressure to urinary albumin excretion differed strikingly between patients with (46%) and without (54%) retinopathy. In those with retinopathy, mean urinary albumin excretion rate was normal (< 20 micrograms/min) below median diastolic pressure (75 mmHg) and increased steeply (p < 0.001) with blood pressure above this level. However, in patients without retinopathy, mean albumin excretion rate was normal across the range of diastolic pressure. This finding could not be explained by differences in glycaemic control or duration of diabetes between patients with and without retinopathy. These data identify a subgroup of patients whose high risk of nephropathy may reflect abnormal renal vulnerability to mildly raised blood pressure. Retinopathy is a close correlate of this vulnerability. Detection of even mild retinopathy, together with raised blood pressure, may be important in assessing nephropathy risk. PMID- 7489846 TI - Albumin excretion and vascular deaths in NIDDM. AB - Non-insulin-dependent diabetes mellitus (NIDDM) is associated with premature mortality, generally thought to be exaggerated in patients with microalbuminuria. This prospective 8-year follow-up study aimed to determine outcome, mortality and cause of death in NIDDM patients with abnormal urinary albumin excretion compared to those with normal albumin excretion. We recruited 153 NIDDM patients with abnormal urinary albumin excretion and 153 control subjects with albumin excretion within the normal non-diabetic range, matched for age, sex and duration of diabetes, from three University hospital diabetic clinics in Newcastle upon Tyne. The outcome measures were status at follow-up, mortality and cause of death. Subjects with abnormal albumin excretion had a significantly higher 8-year mortality than matched control subjects (Odds Ratio 1.47, p = 0.02; 108 vs 66 per 1000 person years follow-up, p < 0.001). This difference was seen at all levels of abnormal albumin excretion, from just outside the normal range (10.6-29.9 microgram/min: 104 vs 61 per 1000 person years follow-up, p < 0.001) to more conventional definitions of microalbuminuria (> or = 30 micrograms/min: 111 vs 71 per 1000 person years follow-up, p < 0.01). Those with abnormal albumin excretion had an excess of vascular deaths compared to matched control subjects (Odds Ratio 1.70, p = 0.009), again at different levels of albumin excretion (10.6-29.9 micrograms/min p < 0.01, 30-150 micrograms/min p < 0.05). On multivariate analysis, age, initial ischaemic heart disease and initial albumin excretion rates were independent predictors of death from all causes.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7489845 TI - Cholesterol-lowering therapy may retard the progression of diabetic nephropathy. AB - There is experimental evidence to suggest that hypercholesterolaemia may play a pathogenetic role in progressive glomerular injury. We investigated the effect of cholesterol-lowering therapy on the progression of diabetic nephropathy in 34 patients with non-insulin-dependent diabetes mellitus. Patients were randomly assigned in a single-blind fashion to treatment with either lovastatin, an HMG CoA reductase inhibitor (n = 16; mean dose 30.0 +/- 12.6 mg/day) or placebo (n = 18) for 2 years. Renal function was assessed by serially measuring the serum creatinine, glomerular filtration rate (using Cr51-EDTA), and 24-h urinary protein excretion. Lovastatin treatment was associated with significant reductions in total cholesterol (p < 0.001), LDL-cholesterol (p < 0.001) and apo B (p < 0.01), the reductions at 24 months being 26, 30 and 18%, respectively. Beneficial effects on serum triglyceride, HDL-cholesterol and apo A1 levels were also observed. Lp(a) showed no significant change in both groups. Glomerular filtration rate deteriorated significantly in the placebo group after 24 months (p < 0.025) but showed no significant change in the lovastatin-treated patients. The increase in serum creatinine was statistically significant (p < 0.02) in placebo-treated patients at 12 and 24 months, and in the lovastatin group after 24 months. Twenty-four hour urinary protein excretion increased in both groups (p < 0.05). Lovastatin treatment was not associated with significant elevations in liver or muscle enzymes. We conclude that effective normalisation of hypercholesterolaemia may retard the progression of diabetic nephropathy. PMID- 7489848 TI - A new marker in the HLA class I region is associated with the age at onset of IDDM. AB - The (MHC) class II association with insulin-dependent diabetes mellitus (IDDM) is well documented. However, it is likely that genes within the MHC class III and the class I region also play a role in determining susceptibility to IDDM. In this study we have used a novel molecular probe to investigate the class I P3A and P3B loci of 179 patients with IDDM and 142 normal control subjects. A highly significant increase in the frequency of the class I P3 4.0;1.5 kilobase (kb) and 4.0;1.8;1.5 kb genotypes was found in patients compared to the control subjects (chi 2 46.8, 6 df, p < 0.0001). The association with the P3B 1.5 kb allele was strongly associated with the age at onset of diabetes, being present in 96.2% of subjects who developed diabetes between the age of 10-20 years compared to 55.0 and 74.6% who developed diabetes before 10 years or after 20 years, respectively (chi 2 31.4, p < 0.0001). There was no evidence for linkage disequilibrium between the DQA1 and DQB1 loci and P3B suggesting that this is an independent association. In conclusion, these results suggest that genes in both the MHC class I and II regions confer susceptibility to IDDM and are related to the age at onset of the disease. PMID- 7489849 TI - Hypomagnesaemia in childhood IDDM and risk of nephropathy. PMID- 7489847 TI - Polymorphisms of the human hexokinase II gene: lack of association with NIDDM and insulin resistance. AB - Skeletal muscle and adipose tissue hexokinase II is a promising candidate gene for non-insulin-dependent diabetes mellitus (NIDDM) and insulin resistance. Therefore, we investigated the association of alleles at four polymorphic loci in this gene with NIDDM and insulin resistance in 110 Finnish diabetic patients with NIDDM and in 97 Finnish control subjects with normal glucose tolerance and a negative family history of diabetes. The four polymorphic nucleotide substitutions (silent) in the coding region of the hexokinase II gene were: GAC 251 GAT (exon 7), AAC 692 AAT and CCG 736 CCC (exon 15), and CTG 766 CTA (exon 16). Allele frequencies of each of these polymorphisms did not differ between patients with NIDDM and control subjects. In addition, subjects who were homozygous for the less frequent allele of each of the four polymorphisms had a similar degree of insulin resistance, as determined by the euglycaemic clamp technique, as did the subjects who were homozygous for the common allele in both control subjects and in patients with NIDDM. In conclusion, polymorphisms in the hexokinase II gene are not associated with the risk of NIDDM or insulin resistance in the Finnish population. PMID- 7489850 TI - Plasma fibrinogen as cardiovascular risk factor. PMID- 7489851 TI - Disordered gastric motor function in diabetes mellitus. PMID- 7489852 TI - Increased expression of type III and type IV collagen in diabetic nephropathy. PMID- 7489854 TI - Health care for all. PMID- 7489853 TI - Assignment of islet cell antibody titres and selection of subjects for IDDM prevention trials. PMID- 7489855 TI - Requests for treatment: ethical limits on cosmetic dentistry. PMID- 7489856 TI - Ceramic shade selection: by whom, where, and under what circumstances. PMID- 7489857 TI - Erythromycin and ketoconazole (Nizoral) associated with terfenadine (Seldane) induced ventricular arrhythmias. PMID- 7489858 TI - Options for treating temporomandibular disorders. PMID- 7489859 TI - Simple apicoectomy. PMID- 7489860 TI - Clinical applications of synthetic bone in dentistry, Part II: Periodontal and bony defects in conjunction with dental implants. PMID- 7489861 TI - Scanning electron microscopic comparison of canal preparation with Nd:YAG laser and hand instrumentation: a preliminary study. PMID- 7489862 TI - Repair of high-copper amalgam with and without an adhesive system: in vitro assessment of microleakage and shear bond strength. PMID- 7489863 TI - Rubber-dam isolation of fixed partial dentures. PMID- 7489865 TI - Why patients leave practices. PMID- 7489864 TI - Periodontic-endodontic treatment of severe alveolar lesions. PMID- 7489866 TI - Intrapocket tetracycline release in periodontal therapy. PMID- 7489867 TI - Myths and rote procedures. PMID- 7489868 TI - Is there a problem? PMID- 7489869 TI - Antibiotics and their use in the treatment of orofacial infections, Part I. PMID- 7489870 TI - Effectiveness of radiographic film barrier envelopes. PMID- 7489871 TI - Uremic stomatitis associated with undiagnosed renal failure. PMID- 7489872 TI - Adverse reaction to ibuprofen overdose. PMID- 7489873 TI - Human herpesviruses: research, and threats to health professionals. PMID- 7489874 TI - C-shaped root canal systems: diagnosis, treatment, and restoration. PMID- 7489875 TI - Proportional length determination: mathematical method. PMID- 7489876 TI - Single-tooth replacement options with the IMZ implant. PMID- 7489877 TI - Screening dentists for risk factors associated with cardiovascular disease. PMID- 7489879 TI - Restoration of facial defects by the prosthodontist. PMID- 7489878 TI - Prevalence of oral lesions in symptomatic and asymptomatic HIV patients. PMID- 7489880 TI - Effectiveness of steam sterilization in killing spores of Bacillus stearothermophilus in prophylaxis angles. PMID- 7489881 TI - Stability of thermoplastic custom impression trays. PMID- 7489882 TI - [Use of the pGB725 probe containing poly-TG-repeats for detecting polymorphic microsatellite loci in the genome of the chicken Gallus gallus domesticus]. AB - Using [32P]-labeled hybridization probe of plasmid pGB725 enriched with poly-TG pairs of nucleotides, chicken genome fingerprints were obtained. High genetic informativity of hybridization profiles and the possibility of polymorphic loci in the telomeric macrochromosomes' regions was revealed. PMID- 7489883 TI - [Expression of a splice-variant of the mts1 gene in normal and tumorous human tissue]. AB - Data on cloning of cDNA corresponding to human mts1 gene transcripts are presented. By comparing nucleotide sequences of the genomic DNA clone and cDNA of mts1, it was shown that human osteosarcoma OHS cells contain two alternative splice variants of mts1 transcripts. Alternative splicing occurs in the 5' untranslated region of the mts1 pre-mRNA. Both splice variants, hu-mts1 and hu mts1(var), demonstrate similar stability in the cells, and each contains one open reading frame for the MTS1 protein. However, the two types of transcripts are translated with different effectiveness. The level of transcription of mts1 splice variants in different normal and neoplastic tissues and cell lines varies significantly. The role of alternative splicing as the mechanism responsible for posttranscriptional regulation of mts1 gene expression is discussed. PMID- 7489884 TI - [Determination and comparative analysis of the nucleotide sequence for minireplicon fragments of the cryptic plasmid p1414 from the soil strain of Bacillus subtilis]. AB - Nucleotide sequences of two minireplicon fragments of the p1414 cryptic plasmid of Bacillus subtilis were determined. The fragments corresponded to the region containing ori(+) and the gene coding for Rep protein. Comparing sequences of the fragments with corresponding sequences of other ss+ plasmids suggested that ori(+) of p1414 belongs to the family of endogenous cryptic B. subtilis plasmids, which form an individual, closely related subgroup in the group of ori(+) sequences of the pC194 type. It was found that the amino acid sequence of a conservative FLTLTV motif located, together with its flanking sequences, at the N ends of Rep proteins encoded by different ss+ plasmids, is similar to those of several transmembrane proteins and signal peptides. These results, together with computer data on predicting the secondary and tertiary structure of the N terminal domain of the p1414 Rep protein, suggest that the domain can serve as a "membrane anchor" during plasmid replication. PMID- 7489885 TI - [Emergence of a recessive lethal mutation in derivatives of an unstable X(Z) chromosome from Drosophila melanogaster]. AB - Xz chromosome isolated from a natural population of Drosophila melanogaster is characterized by spontaneous mutability of the genes yellow, white, and signed and the appearance of chromosomal rearrangements. In mutant lines derived from the line carrying the Xz chromosome that had one, two, or three unstable visible mutations (markers), the rate of appearance of sex-linked lethal mutations was analyzed. This rate was shown to increase with an increase in the number of markers in a line. This phenomenon, termed "marker induction", might explain the phenotypic homogeneity of natural Drosophila populations. Spontaneous lethal mutations were mapped, and their nonrandom distribution along the Xz chromosome was shown. Along with common "hot spot" of lethal mutations, the derivatives for the Xz chromosomes had their own specific sites for lethal mutations. In some cases, the appearance of lethal mutations was accompanied by the formation of inversions in the Xz chromosome. The lethal destabilization of the Xz derivatives, caused by selection for accumulation of visible mutations, is associated with an increase in the number of hot spots for lethal mutations. Presumably, these hot spots are hot sites for the transposition of mobile genetic elements. PMID- 7489886 TI - [Determinants of hybrid dysgenesis in the native population of Drosophila from Altai]. AB - Localization of mobile elements P and hobo in the genomes of isofemale Drosophila lines obtained from a natural population from Biisk (Altai) was analyzed using in situ hybridization. The average copy number per genome was 27.1 for P and 22.0 for hobo. The highest number of P and hobo copies was recorded in the 3R and 21 chromosomes, respectively. The X chromosome contained the lowest number of hobo copies. For P, this relationship was not shown. Both transposons had preferential localization sites, or "hot spots", which partly coincided with intercalary heterochromatin regions. Correlation analysis of P and hobo copy number showed independent distribution of these hybrid dysgenesis determinants. The 1A site, which is thought to be associated with the P-cytotype expression, was not labelled in any line. PMID- 7489887 TI - [Features of ontogenetic development of Drosophila melanogaster in the dysgenetic cross system]. AB - Three families of mobile elements that induce P-M, H-E, and I-R hybrid dysgenesis in Drosophila melanogaster were activated by crossing files of different cytotypes. Manifestation of gonadal sterility in F1 hybrid progeny was dependent on the temperature of development. The systems differed significantly in lethality of F2 hybrids at various stages of ontogeny (embryos, larvae, pupae, and adult flies). The highest embryo lethality was found in the P-M system at the cleavage stage. In the I-R and H-E systems, the peak of embryonic death corresponded to the stages of blastoderm and organogenesis, respectively. Experimental results are discussed in view of molecular and cytological characteristics of interacting strains and existing hypotheses for regulation xof transposition of P, hobo, and I mobile elements. PMID- 7489888 TI - [Determination of a temperature-sensitive period of the new mutation lawc(P1) in Drosophila melanogaster]. AB - Determination of the period of temperature sensitivity in the temperature sensitive allele of the regulatory lawc(P1) mutation was performed. Homeotic transformation of arista into tarsus, frequency of leg deformation, and bristle superexpression were examined. The sensitive periods were detected using reciprocal changes of cultivation temperature from 28 to 17 degrees C and from 17 to 28 degrees C. The temperature-sensitive period (TSP) for arista transformation was shown to manifest polyphasic expression and sexual dimorphism. In females, it occurred in late third instar larvae (the first phase) and prepupae (the second stage); in males, it includes the whole period from the late third instar larva upto and including prepupa. TSP for the frequency of deformed legs was polyphasic and took place during the third larval instar (the first phase) and prepupa stage (the second one). TSP for bristle superexpression occurred during a single interval from the late third larval instar until the early prepupa stage. The products of the lawc gene are assumed to play a role both in the cell proliferation in the legs of antennal and imaginal discs and in the control of bristle expression at the final stages of Drosophila ontogeny. PMID- 7489889 TI - [Analysis of the variable structure of a cluster of ribosomal genes in the Insecta class]. AB - General estimation of ribosomal DNA variation within the Insecta class is presented. It is shown that, using blot-hybridization, one can detect differences in the structure of the ribosomal gene cluster not only between genera within an order, but also between species within a genera, including sibling species. Structure of the ribosomal gene cluster of the Coccinelidae family (ladybirds) is analyzed. It is shown that cloned highly conservative regions of ribosomal DNA of Tetrahymena pyriformis can be used as probes for analyzing ribosomal genes in insects. PMID- 7489890 TI - [DNA polymorphism of the BoLA-DRB3 gene in cattle in connection with resistance and susceptibility to leukemia]. AB - Polymorphism of exon 2 of the BoLA-DRB3 gene was investigated by the PCR-RFLP method in a sample of healthy and leukemia-afflicted Black Pied cattle. Allele variety was studied and allele frequencies were determined in a total sample and in the two groups. Alleles mediating resistance (BoLA-DRB3.2*11, *23, and *28) and susceptibility to leukemia (DRB3.2*22, *24, *16, and *8) were revealed in Black Pied cattle. The dominant type of inheritance of the disease resistance was confirmed. On the basis of original and published data obtained earlier for Holstein-Friesian cattle, a conclusion was made about the universal character of the spectrum of BoLA-DRB3 alleles providing resistance and susceptibility to leukemia. PMID- 7489891 TI - [Genetic-demographic study of mountain populations from Dagestan and their migrants to the lowlands. Comparison of basic parameters of fitness]. AB - Results of a total genetic demography study of females of postreproductive age from both parts of "splits" highland isolates of Dagestan, highlanders, and migrants to lowlands are described. The components of natural selection that were related to differential mortality and differential fertility were estimated in the highlanders and the migrants using Crow indices. Offsprings of female migrants were characterized by a higher prenatal (spontaneous abortions and still births) and a lower prereproductive mortality. These differences may be accounted for by two mechanisms: (1) the effects of changing marital structure (female migrants are more exogamous) and (2) better medical care for the children of migrants compared to those living in highland auls (settlements), which are still difficult to access. It was demonstrated that mortality of probands' children before reproductive age, as well as the proportion of their close genetic relatives that died within five years after resettling, increased with an increase in the level of individual inbreeding in women examined (probands). These data allow us to suggest that the drastic increase revealed in mortality of the highlanders during the first years after resettling may be partially attributed to high levels of inbreeding and heterozygosity; these, in turn, decrease the individual's nonspecific resistance to new ecological factors. Fertility and prenatal mortality appeared to increase and decrease, respectively, with an increase in the inbreeding level. PMID- 7489892 TI - [Polymorphism of mitochondrial DNA region V in native and migrant inhabitants of Northeast Asia]. AB - Data on polymorphism of the V region of mitochondrial DNA (mtDNA) in eastern Slavs from Magadan (N = 108), Asian Eskimos (N = 21), and coastal Chukchi (N = 10) are presented. The region V insertion-deletion polymorphism was shown to be absent in the samples of native and immigrant population of northeastern Asia. The Hae III-Ava II restriction polymorphism of the 8249-8254 bp part of this mtDNA region was found only in eastern Slavs. In them, the variant Hae III(-)/Ava II(+) and the novel type of polymorphism Hae III(-)/Ava II(-) were recorded at frequencies 2.8% and 1.85%, respectively. The low level of diversity of mtDNA types in Arctic populations of the world is discussed. A hypothesis is advanced that the low level of mtDNA diversity in Arctic population of humans is determined by several factors. These include an initially low number of mtDNA types in forming ancestral populations, genetic drift, isolation, and continuous adaptation of the populations. The latter is associated with selection of genotypes optimal for living in the extreme climatic and geographic conditions of the North; some of these genotypes control a low level of free oxygen radicals in the cell. PMID- 7489894 TI - An investigation of conservation-biased gap-penalties for multiple protein sequence alignment. AB - Sequence conservation in a multiple sequence alignment (or profile) is often used to influence the alignment of further sequences onto the profile. Most methods, however, have considered only the opening of a gap at a single point and not what is contained in the inserted segment of one sequence (or profile) or what terminates the 'broken' ends of the other. An alignment algorithm is described that incorporates these aspects and the relative importance of the contribution from the insert and the 'broken' ends has been assessed. The approach was tested on families of very remotely related sequences using a novel protocol that was developed to quantify both the stability and generality of the solution. PMID- 7489893 TI - [Chromosomal mosaicism of extraembryonic cell membrane, detected during prenatal diagnosis]. AB - Data on detection of chromosomal mosaicism in amnionic cells and chorionic villi obtained by prenatal cytogenic diagnosis are presented. The frequency of chromosomal mosaicism in preparations of amniotic fluid cell culture was 2.6% (6 out of 226), and that in "direct" villus preparations was 1.6% (13 out of 774). The necessity perform an additional analysis of other fetal cells or neonatal lymphocytes to specify the diagnosis was shown. The analysis of the outcome of pregnancies during which chromosomal mosaicism in the extraembryonic cells was detected indicates that these women from a high-risk group of at, both genetically and obstetrically: in only 8 out of 19 cases did pregnancies end in normal deliveries at term; in three cases, spontaneous abortions occurred at 16 31 weeks of gestation; in three cases, the pregnancies were terminated due to fetal chromosomal aberrations in nonmosaic form; the outcome of pregnancy in five cases was preterm delivery of an underweight newborn. PMID- 7489895 TI - Analysis of a Bacillus subtilis genome fragment using a co-operative computer system prototype. AB - Analysis of the huge volume of data generated by large scale sequencing projects requires the construction of new, sophisticated computer systems. These systems should be able to manage the biological data as well as the results of their analysis. They should also help the user to choose the most appropriate methods, and to string them together in order to solve a global analysis task. In this paper we present the prototype of a software system providing an environment for the analysis of large-scale sequence data. As a first step toward this end, this environment has been put to the test within the Bacillus subtilis genome sequencing project. This system integrates both the descriptive knowledge of the entities involved (genes, regulatory signals and the like) and the methodological knowledge comprising an extensible set of analytical methods. A knowledge representation based on two existing object-oriented models is used to implement this integrated system. In addition, the present prototype provides a suitable user interface both for displaying simultaneously the results generated by several methods and for interacting with the objects. We present in this paper the analysis of a B. subtilis genome fragment, present in data libraries but not annotated. Annotation of the genes present in the fragment allowed us to combine the results of several methods used for predicting coding sequences, and to characterize it as comprising a cryptic phage, the skin element. Comparison between the annotation of the skin element and a standard region of the chromosome indicated that local features of the nucleotide sequence could discriminate between phage and non-phage DNA sequence. PMID- 7489896 TI - Identification of Campylobacter jejuni and C. coli using the rpoB gene and a cryptic DNA fragment from C. jejuni. AB - Campylobacter jejuni (Cj) and C. coli (Cc) clinical isolates, obtained from three different sources, were characterized using two Cj DNA probes, CJ01 and CJ02. These probes were selected at random by virtue of their stability in Escherichia coli (Ec). CJ01 hybridized specifically with DNA from Cj reference strains, but not with DNA from Cc, C. lari (Cl) nor C. fetus (Cf) reference strains. Using clinical isolates characterized by genome-genome hybridization and biotype, CJ01 hybridized with DNA derived from all Cj strains. However, DNA from four out of ten Cc strains, from three different sources, also hybridized with CJ01, suggestive of this region being heterogeneous between clinical isolates of both species. The nucleotide sequence analysis of CJ01 reveals two incomplete open reading frames (ORFs) that did not show significant homology with any other known sequences. CJ02 hybridized specifically with DNA from Cj and Cc reference strains, but not with DNA from Cl and Cf reference strains. The specificity and sensitivity were maintained upon hybridization with DNA from 31 clinical isolates. CJ02 has an uninterrupted ORF whose deduced amino-acid sequence showed extensive homology with the central region of the Ec and Salmonella typhimurium (St) RNA polymerase beta subunits (52 and 66% similarity, respectively). The most conserved segments correspond to putative functional domains. PMID- 7489897 TI - Characterization of a new DNA polymerase from Schizosaccharomyces pombe: a probable homologue of the Saccharomyces cerevisiae DNA polymerase gamma. AB - Mitochondrial (mt) DNA replication is carried out by the nuclear-encoded DNA polymerase-gamma (Pol-gamma). We have cloned a new DNA polymerase-encoding gene from Schizosaccharomyces pombe (Sp), which we believe encodes the homologue of the Saccharomyces cerevisiae (Sc) mt DNA polymerase (MIP1). The putative Sp pol gamma gene expressed a transcript of approx. 4-kb that contained a 3-kb open reading frame encoding a polypeptide of 1018 amino acids (aa) (116 kDa). This Sp Pol-gamma is 48% identical to the Sc MIP1 and contains uniquely conserved regions not found in the bacterial PolI-type DNA polymerases. The most notable difference between these two proteins is that the MIP1 product has a 236-aa C-terminal region beyond motif C that is not found in Sp Pol-gamma. Chromosomal mapping and genomic sequencing of the Sp pol gamma places this gene on chromosome III downstream from the triose phosphate isomerase-encoding gene. PMID- 7489898 TI - Cloning and sequencing of the beta-glucosidase-encoding gene from Candida molischiana strain 35M5N. AB - We have isolated a gene (bgln) encoding beta-glucosidase (beta Glu) from a cosmid library of the yeast, Candida molischiana 35M5N. The nucleotide sequence of bgln and its flanking regions was determined. This gene was found to be composed of 2289 bp and 763 amino acid (aa) residues encoding an 83.3-kDa protein. The aa sequence shared eleven putative N-glycosylation sites. Homology comparisons showed that this enzyme can be considered as a new member of the family-3 glycosyl hydrolases. Multiple alignment experiments revealed four conserved regions on aa sequences from beta Glu of this family. PMID- 7489899 TI - Cloning of the Candida albicans HIS1 gene by direct complementation of a C. albicans histidine auxotroph using an improved double-ARS shuttle vector. AB - ARS2 and ARS3 are two Candida albicans (Ca) DNA fragments with autonomous replicating activity that have been shown to promote non-integrative genetic transformation of both Ca and Saccharomyces cerevisiae (Sc). We have developed several shuttle vectors based on either ARS fragment, or the combination of both, and using the CaURA3 gene as a selection marker. The combination of ARS2 and ARS3 fragments in a single vector did not increase transformation frequencies but improved the stability of transformant plasmids in Ca cells, so that the degree of intracellular recombination was reduced. A Ca genomic DNA library was constructed on the double-ARS vector, pRM1, to be used for direct cloning in Ca by complementation of the histidine auxotrophy of strain CA9. By screening this library, we cloned CaHIS1, the Ca gene that encodes ATP phosphoribosyl transferase, one of the enzymes that participates in histidine biosynthesis. The deduced protein, CaHis1p, is 60.6% identical (73% similar) to ScHis1p (EC 2.4.2.17). The cloned gene is the first auxotrophic gene marker mapped to fragment I of chromosome 5 in the standard Ca genetic map. Our results represent the first demonstration of a direct cloning system in the opportunistic fungus Ca that does not require the use of an intermediate host such as Sc for plasmid rescue. This system could be used for the isolation of any gene affected in Ca mutants displaying a selectable or identifiable phenotype. PMID- 7489900 TI - Cloning and characterization of the cDNAs and genes (mep20) encoding homologous metalloproteinases from Aspergillus flavus and A. fumigatus. AB - Aspergillus fumigatus (Afu) and A. flavus (Afl), two causative agents of invasive aspergillosis, produce highly homologous serine proteinases. In addition, the former produces a 42-kDa metalloproteinase (MEP), whereas the latter produces a 23-kDa MEP. The cDNA and the gene encoding the 42-kDa MEP were cloned and sequenced. Here, we report the cloning of the cDNA and the gene encoding the 23 kDa MEP from Afl and a homologous gene from the Afu. Using degenerate primers based on the amino acid (aa) sequence of A. oryzae (Ao) MEP and thermolysin-like proteinases, a 282-bp fragment of the 23-kDa MEP-encoding gene of Afl was cloned by PCR. A 6.5-kb KpnI fragment of Afl genomic DNA containing the complete gene was cloned. The open reading frame (ORF) in this gene encodes a protein of 381 aa. Since the mature enzyme from this and other aspergilli would have a theoretical molecular mass of about 20 kDa, this MEP-encoding gene is designated mep20. A Western blot of the protein in the culture filtrate of Afl with polyclonal antibodies prepared against the MEP showed a single band at 23 kDa. The N-terminal sequence of the extracellular MEP20, TKVAS, was found at aa 194 198 within the ORF. Thus, the primary translation product has a putative 19-aa signal and a pro region of 174 aa. A homologous gene cloned from a genomic DNA library of Afu showed an ORF encoding 365 aa. Comparison of the nucleotide (nt) sequences of the cDNAs cloned by RT-PCR with their respective genes showed that there are no introns in the ORF of mep20 in Afl, but there is a 59-bp intron in the gene from Afu. The MEP20 of Afl and Afu have 68% identity and show weak immunological cross reactivity. MEP20 from both these fungi share about 60% sequence identity with the penicillolysin of Penicillium citrinum and the neutral protease II of Ao. MEP20 of Afl and Afu show only the conserved sequence, HEFTHA, but not the two other conserved sequences seen in thermolysins and similar MEP. PMID- 7489901 TI - Green fluorescent protein production in the cellular slime molds Polysphondylium pallidum and Dictyostelium discoideum. AB - The green fluorescent protein-encoding gene from Aequorea victoria has been cloned into several different transforming vectors and expressed in the cellular slime molds, Polysphondylium pallidum and Dictyostelium discoideum. We find that the protein is stable and non-toxic in both species, can be easily visualized in living and fixed specimens, and can be used to purify rare cells by fluorescence activated cell sorting (FACS). PMID- 7489902 TI - Structure of the Leptomonas seymouri trans-spliceosomal U2 snRNA-encoding gene; potential U2-U6 snRNA interactions conform to the cis-splicing counterpart. AB - We have characterized the U2 small nuclear RNA (snRNA)-encoding gene from the monogenetic trypanosomatid, Leptomonas seymouri (Ls), to begin to identify the RNA-RNA interactions that direct trans-splicing in kinetoplastid protozoa. The U2 gene, which is single copy in this organism, was isolated and sequenced. Although the Ls U2 snRNA contains many of the sequence and secondary structure elements that are conserved among the U2 snRNAs of cis-splicing organisms, it lacks the stem-loop III region and the intron branch point-recognition region, as do other trypanosomatid U2 snRNAs. A transcriptional promoter element within the Trypanosoma brucei U2 gene [Fantoni et al., Mol. Cell. Biol. 14 (1994) 2021-2028] is conserved in the homologous Ls gene. A crucial step in cis-splicing reactions involves specific base-pairing interactions between the U2 and U6 snRNAs. We show here that in trypanosomatids, where no cis-splicing occurs, these same interactions are possible. This highlights key similarities between the two RNA processing events. PMID- 7489903 TI - A method for preparing M13 or pUC libraries for sequencing DNA with high G + C or A + T contents. AB - A simple method is described to generate M13 or pUC libraries from DNA with a very high G + C or A + T content. The G + C-rich DNA is partially digested with HinPI or HpaII restriction enzymes and cloned into the vector linearized in its multiple cloning site with AccI. The A + T-rich DNA is partially digested with TspI and cloned into the EcoRI-linearized vector. These libraries are suitable for large-scale DNA sequencing. PMID- 7489904 TI - A phage display vector with improved stability, applicability and ease of manipulation. AB - We modified a combinatorial library display vector, pCOMB3, to provide a stable, easily manipulated, high-copy vector for the display of a random hexapeptide library. The propensity of the original phagemid to accumulate 800-1000-bp deletions in the region of the cloning site has been eliminated. Furthermore, the small 63-bp 'stuffer' at the cloning site was replaced with a 2114-bp DNA fragment from adenovirus 2. This produced a 5808-bp vector, that we have named pICD1LS, with the appropriate characteristics for single-peptide phage display. Libraries of greater than 10(6) molecules were produced with this vector. PMID- 7489905 TI - New plasmids carrying antibiotic-resistance cassettes. AB - A series of new plasmid vectors is described that carry gene cassettes imparting resistance to the antibiotics chloramphenicol (CmR), kanamycin (KmR), tetracycline (TcR) and spectinomycin/streptomycin (Sp/SmR). The gene cassettes are symmetrically flanked by several restriction sites. In addition, several restriction sites that are normally found internal to the gene cassettes have been eliminated, thereby expanding the number of restriction enzymes available to excise an intact antibiotic-resistance gene. The gene cassettes are carried by high-copy-number plasmids that confer ampicillin resistance (ApR). PMID- 7489906 TI - An IS903-based vector for transposon mutagenesis and the isolation of gene fusions. AB - A derivative of the IS903 transposon (Tn) is described that is capable of creating lacZ gene fusions upon transposition. It should find wide use as a tool for Tn mutagenesis in bacteria since it can be used both to generate mutants and to examine gene expression. The transposase-encoding gene (tnp) is located outside the Tn in the vector, thus Tn insertions into a genome are stably maintained in the absence of its cis-acting transposase (Tnp). The element carries a KmR gene allowing for the direct selection of transposition events in hosts that cannot support pBR322 plasmid replication and facilitating the subcloning of genes into which the Tn has inserted. PMID- 7489908 TI - Identification of the gene encoding a mechanosensitive channel MscL homologue in Clostridium perfringens. AB - The mscL gene, which encodes the protein forming a large-conductance mechanosensitive channel (MscL) in Escherichia coli, has previously been cloned and sequenced by Sukharev et al. [Nature 368 (1994) 265-268]. We found a gene homologous to mscL in Clostridium perfringens which is located just downstream from the collagenase-encoding gene in the opposite direction. PMID- 7489907 TI - A novel Escherichia coli lipoprotein expression vector. AB - A novel Escherichia coli (Ec) lipoprotein expression plasmid, pSJLP, was constructed. The plasmid contains a truncated alkaline phosphatase gene (phoA) located downstream from the Lac repressor gene lacIq and the IPTG inducible Ptac promoter. The phoA gene was truncated by deleting the native phoA signal sequence and fusing the truncated phoA gene to the lipoprotein signal sequence of the major Ec lipoprotein LPP. The recombinant LPP::PhoA fusion protein is produced and processed as a lipoprotein and can therefore be used as substrate for a novel signal peptidase II assay. PMID- 7489909 TI - New multifunctional Escherichia coli-Streptomyces shuttle vectors allowing blue white screening on XGal plates. AB - Four new shuttle vectors for Escherichia coli (Ec) and Streptomyces, pUWL218, pUWL219, pUWL-SK and pUWL-KS, which permit recognition of recombinant (re-) plasmids on XGal plates in Ec, were constructed. These vectors contain the replication functions of the Streptomyces wide-host-range multicopy plasmid pIJ101, the tsr gene conferring resistance to thiostrepton in Streptomyces, the ColEI origin of replication from the pUC plasmids for replication in Ec and the bla gene conferring resistance to ampicillin in Ec. They possess multiple cloning sites with a number of unique restriction sites and allow direct sequencing of re derivatives using the pUC sequencing primers. PMID- 7489910 TI - Genetic organization of a small cryptic plasmid of Helicobacter pylori. AB - A 2.9-kb cryptic plasmid of Helicobacter pylori (Hp), pHel1, was isolated and the complete nucleotide (nt) sequence was determined. An open reading frame (ORF1) was identified encoding a putative polypeptide of 63,709 Da, the existence and correct size of which was confirmed by T7 promoter expression analysis. The ORF1 sequence showed strong amino-acid sequence identity to a recently identified putative ORF1 protein of a cryptic Hp plasmid, pHPM180, and significant homologies to putative Rep proteins of Campylobacter coli (RepB) and Pediococcus halophilus (RepA), and was therefore designated RepA. A functional role of RepA in replication of pHel1 was demonstrated by the fact that only pHel1 plasmid derivatives with an intact repA gene were able to autonomously replicate in Hp. Upstream of repA, a 22-bp sequence was recognized which was tandemly repeated four and a half times, a feature typical for many replication origins (ori) and commonly termed a DNA iteron. Analysis of the repA upstream region by primer extension identified a transcription start point for the repA mRNA, but did not correspond to known consensus promoter sequences. Southern hybridizations using pHel1 as a probe under stringent conditions revealed that homologous sequences to pHel1 were present in nearly all plasmid-carrying Hp strains, but not in a plasmid-carrying Helicobacter felis strain, suggesting that this type of replicon is predominantly found in the Hp species. PMID- 7489911 TI - Cloning and characterization of the Saccharomyces cerevisiae SVS1 gene which encodes a serine- and threonine-rich protein required for vanadate resistance. AB - A novel Saccharomyces cerevisiae (Sc) SVS1 gene was cloned as a multicopy suppressor of vanadate (Vn) sensitivity (VnS) due to a calcineurin (CaN) null mutation. SVS1 encoded a 260-amino-acid protein abundant in Ser and Thr residues, with a putative signal sequence at the N terminus. Deletion of SVS1 resulted in increased sensitivity to Vn, but not to other metallic ions or drugs. Northern analysis of the SVS1 mRNA indicated that the induction of the gene occurred specifically in the response to Vn. These results suggested that Sc has a mechanism to enhance the tolerance to Vn by increasing the expression of SVS1. The results of genetic experiments suggested that CaN and the Svs1 proteins act in separate pathways to enhance the tolerance to Vn. PMID- 7489912 TI - Identification of differentially expressed Leishmania donovani genes using arbitrarily primed polymerase chain reactions. AB - Arbitrarily primed polymerase chain reactions (AP-PCR) were used to amplify polymorphic DNA fragments from the genomes of a variety of geographic isolates of Leishmania donovani (Ld). From the latter, five polymorphic DNA fragments were cloned and sequence analysis identified 15 unique clones. Northern blot analysis showed that 13 of the 15 clones hybridized to transcribed RNAs isolated from Ld. Eight of these 13 AP-PCR clones specifically hybridized to Ld RNAs that were differentially expressed in promastigote and 'amastigote' cells. Comparative Northern analysis of four differentially expressed AP-PCR clones indicated that two clones, LdS-14-14 and LdI-9-7, were expressed in Ld and several other Leishmania species. However, RNAs corresponding to two other AP-PCR clones, LdE-6 1 and LdI-9-5, were detected only in members of the Ld complex, and not in L. major (Lm) or L. tropica (Lt). Comparative Southern blot analysis of the LdS-14 14 locus revealed numerous restriction-fragment length polymorphisms (RFLP) distinguishing Lm and Lt from the Ld isolates and L. infantum. However, the LdS 14-14 loci were mapped to similar-sized chromosomes observed among all Old World Leishmania species tested, indicating that localized nucleotide divergence, not chromosomal rearrangement, was responsible for altered Southern blot patterns. These results demonstrate that AP-PCR is a very useful method for identifying expressed gene sequences in organisms of relatively low-complexity genomes. Interestingly, the majority of these sequences identified in this study correspond to differentially expressed genes.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7489913 TI - A growth-defective kirromycin-resistant EF-Tu Escherichia coli mutant and a spontaneously evolved suppression of the defect. AB - This study has investigated the cause of a growth-defect phenotype of a mutation in the elongation factor EF-Tu from Escherichia coli. An M13-based genetic retrieval system reported by Zeef and Bosch [Mol. Gen. Genet. 238 (1993) 252-260] was used to segregate and identify an extremely growth-defective kirromycin resistant (KrR) tufA mutation, encoding Gln124-->Lys (Q124K), from a KrR parent strain. This original strain also contained mutations, 124com1 and 124com2, that appear to have evolved to suppress the Q124K tufA mutation. In this communication we present these M13-based genetic experiments together with additional genetic and protein characterization experiments to clarify the basis of this complementation. The data indicate that the serious growth defect of Q124K originates from a defective GTP/GDP interaction. The GTP/GDP binding and GTP hydrolysis characteristics of ET-Tu Q124K were different from wild-type EF-Tu and especially of another KrR EF-Tu mutant A375T. In line with this, 124com1 specifically complemented EF-Tu Q124K, whereas the growth defects of strains containing EF-Tu mutated at aa 375 were aggravated. We also show that strains containing the segregated tufA Q124K mutation formed filaments. PMID- 7489914 TI - The 3'-5' exonuclease site of DNA polymerase III from gram-positive bacteria: definition of a novel motif structure. AB - The primary structure of the 3'-5' exonuclease (Exo) site of the Gram+ bacterial DNA polymerase III (Pol III) was examined by site-directed mutagenesis of Bacillus subtilis Pol III (BsPol III). It was found to differ significantly from the conventional three-motif substructure established for the Exo site of DNA polymerase I of Escherichia coli (EcPol I) and the majority of other DNA polymerase-exonucleases. Motifs I and II were conventionally organized and anchored functionally by the predicted carboxylate residues. However, the conventional downstream motif, motif III, was replaced by motif III epsilon, a novel 55-amino-acid (aa) segment incorporating three essential aa (His565, Asp533 and Asp570) which are strictly conserved in three Gram+ Pol III and in the Ec Exo epsilon (epsilon). Despite its unique substructure, the Gram+ Pol III-specific Exo site was conventionally independent of Pol, the site of 2' deoxyribonucleoside 5-triphosphate (dNTP) binding and polymerization. The entire Exo site, including motif III epsilon, could be deleted without profoundly affecting the enzyme's capacity to polymerize dNTPs. Conversely, Pol and all other sequences downstream of the Exo site could be deleted with little apparent effect on Exo activity. Whether the three essential aa within the unique motif III epsilon substructure participate in the conventional two-metal-ion mechanism elucidated for the model Exo site of EcPol I, remains to be established. PMID- 7489915 TI - Characterization and overexpression of the gene encoding Staphylococcus aureus DNA polymerase III. AB - The polC gene specifying DNA polymerase III (PolIII) of Staphylococcus aureus (Sa), was cloned with a novel strategy and found to contain a 4305-bp open reading frame (ORF) encoding a polypeptide of approx. 162 kDa. The 1435-codon ORF was engineered into an Escherichia coli (Ec) expression plasmid under the control of the lac promoter and its repressor. Derepression of Ec transformants carrying the recombinant (re-) vector generated high-level synthesis of active re-Sa PolIII. The re-PolIII was purified to > 98% homogeneity and was shown by N terminal amino acid sequence analysis to be the bona fide product of the Sa polC ORF. The physical and catalytic properties of re-Sa PolIII and its responsiveness to inhibitors of the HPUra type were generally similar to those of Bacillus subtilis (Bs) PolIII. Comparative analysis of the primary structures of Sa PolIII, Bs PolIII and Mycoplasma pulmonis PolIII indicated strong conservation of essential catalytic domains and a novel zinc-finger motif. Comparison of the primary structures of Ec PolIII and these three Gram+ enzymes revealed a region of novel homology and reinforced the likelihood of a specific evolutionary relationship between PolIII of Gram+ and Gram- eubacteria. The polC gene mapped between omega 1074 [Tn551] and recA/ngr on the Sa NCTC 8325 genome. PMID- 7489916 TI - Cloning, sequence analysis and expression of the group A streptococcal guaB gene encoding inosine monophosphate dehydrogenase. AB - Inosine monophosphate dehydrogenase (IMPDH) is an essential enzyme in the biosynthesis of purines. We cloned a group A streptococcal (GAS) DNA fragment containing an open reading frame similar to other bacterial guaB genes encoding IMPDH. The GAS guaB consists of 1479 nucleotides encoding a protein of 493 amino acids. Expression of the GAS guaB in an Escherichia coli guaB mutant restored IMPDH activity, confirming the function of the gene product and demonstrating that the GAS enzyme is active in a heterologous bacterial host. Restriction mapping and Southern hybridization analysis of GAS chromosomal DNA localized guaB to a site approximately 5 kb from the hasA and hasB genes which encode enzymes necessary for hyaluronic acid capsule synthesis. PMID- 7489917 TI - A targeted mutagenesis system for Actinobacillus pleuropneumoniae. AB - We describe methods for the mutagenesis of cloned Actinobacillus pleuropneumoniae (Ap) genes and for the construction of Ap mutants by allelic exchange. We used these methods to construct isogenic mutants of Ap which no longer synthesized a 48-kDa outer membrane protein (AopA). The native aopA locus was replaced with a mutated locus that had been inactivated by insertion of a gene (KmR) encoding kanamycin resistance from Tn903. The inactivated aopA locus was cloned into a conjugative, R6K-derived, lambda pir-dependent suicide vector and introduced into Ap using a filtermating technique. Southern and Western blot analyses indicated that the wild-type locus was replaced by the mutated locus through either single- or double-crossover events, and that AopA was no longer produced by either type of mutant. These methods were used successfully to construct AopA- mutants in Ap serotypes 1 and 5. These methods should be generally useful in constructing mutant loci which can be used to analyze the roles of various Ap genes in the pathogenesis of contagious pleuropneumonia in swine. PMID- 7489918 TI - Genomic organization of the mycobacterial sigma gene cluster. AB - We have previously described sigma A and sigma B and their structural genes, mysA and mysB, respectively, in Mycobacterium smegmatis. We have now sequenced the corresponding regions in the M. tuberculosis and M. leprae chromosomes, and have found the two homologous genes. The chromosomal linkage and the deduced amino acid (aa) sequences of the two genes show very high similarity in the three species of mycobacteria. We also report the finding of two other open reading frames (ORF) in these clusters. orfX, which has an unknown function, is located between mysA and mysB. The other ORF, located downstream from mysB, encodes a homolog of DtxR, the iron regulatory protein from Corynebacterium diphtheriae (Cd). PMID- 7489919 TI - Cloning, sequence analysis, overproduction in Escherichia coli and enzymatic characterization of the RNase HI from Mycobacterium smegmatis. AB - Activity gel analysis of cell extracts from slow- and fast-growing mycobacteria confirmed the presence of several RNase H activities in both classes of organism. The rnhA gene from Mycobacterium smegmatis (Ms) was subsequently cloned using an internal gene segment probe [Mizrahi et al., Gene 136 (1993) 287-290]. The gene encodes a polypeptide of 159 amino acids that shares 50% identity with the RNase HI from Escherichia coli (Ec). However, unlike its counterparts from Gram- bacteria, Ms rnhA does not form an overlapping divergent transcriptional unit with dnaQ (encoding the epsilon (proofreading) subunit of DNA polymerase III). Ms RNase HI was overproduced in Ec as an enzymatically active maltose-binding protein (MBP) fusion protein which cleaved the RNA strand of an RNA.DNA hybrid with a similar site selectivity to that of its Ec homologue. PMID- 7489920 TI - Cloning, sequencing and expression in Escherichia coli of a Streptomyces aureofaciens gene encoding glyceraldehyde-3-phosphate dehydrogenase. AB - The structural gene (gap) encoding glyceraldehyde-3-phosphate dehydrogenase (GAPDH; EC 1.2.1.12) from Streptomyces aureofaciens (Sa) has been cloned and sequenced. The predicted gap product consists of 332 amino acids (aa) (35,312 Da), and has considerable homology (up to 52% aa identity) with other bacterial and eukaryotic gap genes. Sequence analysis of the regions flanking gap revealed two incomplete open reading frames encoding proteins similar to the AraC family of bacterial transcriptional regulators and delta (5)-3-ketosteroid isomerase. The Sa gap gene was expressed at a high level in Escherichia coli (Ec). Transformation of the Ec strain resulted in an up to eightfold increase in specific GAPDH activity. PMID- 7489921 TI - Characterization of the unlinked 16S rDNA and 23S-5S rRNA operon of Wolbachia pipientis, a prokaryotic parasite of insect gonads. AB - The rRNA-encoding genes (rDNAs) have been cloned and characterized from Wolbachia pipientis (Wp), the gonadial bacteria-like parasite of the mosquito Culex pipiens (Cp) and the moth Ephestia cautella (Ec). In Wp from both insect species the rDNAs are organized in a way which appears to be very unusual. The rRNAs are encoded by two unlinked transcription units, each present in a single copy per genome. One contains the 16S rDNA only, while the other is an operon encoding both the 23S and 5S rDNAs. Each transcription unit contains two putative upstream promoters, and downstream a Rho-independent terminator. The 16S rDNA, as well as the 23S-5S rRNA operon are not linked to any tRNA-encoding sequence and lack the antitermination boxes which are usually present immediately downstream from eubacterial promoters of rDNAs. Wp infecting Ec and Cp are highly similar taking as criteria the rDNAs and their flanking sequences. However, it clearly appears that each insect species harbours a different and specific Wp strain, or even subspecies. Phylogenetic relationships deduced from the complete sequences of their rDNAs undoubtedly confirm that Wp from Cp and Ec belong to the alpha-group of Proteobacteria, and are closely related to the Rickettsia. PMID- 7489922 TI - Conservation and variability in Archaea: protein antigens with tandem repeats encoded by a cluster of genes with common motifs in Methanosarcina mazei S-6. AB - Three open reading frames, orf492, orf375 and orf783, were identified in a 5.9-kb DNA fragment from the genome of Methanosarcina mazei S-6 that code for proteins recognized by antibodies against cell-surface antigens. The deduced amino-acid (aa) sequences of orfs492 and 375, i.e., ORF492 and ORF375, contain seven and four copies of an approx. 42-aa repeat, respectively. The aa sequence of ORF783 contains nine copies of an approx. 85-aa repeat, one of which is also present once in each of the first two ORFs. The organization of the repeats is similar to that of some Gram+ cell-wall-associated proteins. Comparative analyses of aa sequences, compositions and hydropathy profiles of the archaeal ORFs showed similarity with surface (S-) layer and outer-membrane proteins of Bacteria and Archaea. PMID- 7489923 TI - The isolation of lactococcal promoters and their use in investigating bacterial luciferase synthesis in Lactococcus lactis. AB - 18 different promoter elements, encompassing a 71-fold range of activity, were isolated from the chromosome of Lactococcus lactis (Ll) MG1363 and from an uncharacterised small isometric bacteriophage of Ll. The Vibrio fischeri (Vf) luciferase-encoding gene (lux) was used as a reporter in Ll, so that the promoters could be identified strictly on the basis of their activity in the homologous host. Sequence and primer extension analysis of six of the promoters has provided a new consensus sequence for the -35 and -10 hexanucleotide motifs present upstream from lactococcal transcription start points. When the nucleotide sequence of the most active promoter (P15) was compared with that of the highly expressed Ll usp45 gene, a novel 8-bp region of homology was identified which corresponded to the newly derived consensus -35 sequence element; this element may therefore be of general importance in Ll gene expression. The isolation of these promoters has also enabled us to investigate the characteristics of the Vf Lux activity in Ll under different physiological conditions using promoters of different strengths. Lux activity in Ll is critically dependent upon the phase of cell growth. Luminescence falls sharply in stationary phase, possibly due to a lack of FMNH2. In contrast to the kinetics of Lux function in Escherichia coli (Ec), Lux activity in Ll declines rapidly after addition of the substrate; the rate of decay is dependent both on the growth phase and on the strength of the promoter. It is apparent that the previously reported thermal instability of Lux is in fact a function of the host organism in which Lux is expressed.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7489924 TI - pWITCH: a versatile two-hybrid assay vector for the production of epitope/activation domain-tagged proteins both in vitro and in yeast. AB - We describe the construction of a new vector, pWITCH, designed to facilitate the characterisation of proteins encoded by novel cDNAs isolated using either a one- or two-hybrid assay. Expression of directionally cloned cDNAs is directed in vivo in Saccharomyces cerevisiae from the inducible GAL10 promoter and in vitro from the T7 promoter, while translation of the expressed cDNAs results in proteins which are tagged in vitro with a specific epitope and in vivo with both the epitope and the VP16 transcription activation domain. The principle of using multiple promoters each able to operate under different conditions to express different combinations of protein domains without the need for subcloning should be generally applicable. PMID- 7489925 TI - Isolation and characterization of the Saccharomyces cerevisiae SUT1 gene involved in sterol uptake. AB - A new gene (SUT1) of Saccharomyces cerevisiae, implicated in sterol uptake, was isolated from a yeast genomic library constructed in a high-copy-number vector by virtue of conferring resistance to fenpropimorph in medium supplemented with ergosterol. The high expression of SUT1 in sterol auxotrophic mutant strains alleviates the requirement for accessory mutations affecting heme biosynthesis and allows sterol uptake in aerobiosis. Measurements of [14C]cholesterol uptake confirmed that SUT1 is involved in sterol absorption. Within the 4.1-kb insert isolated, the functional gene was localised on a 1.7-kb DNA fragment. The nucleotide sequence encodes a predicted protein of 299 amino acids. Northern blot analysis revealed that SUT1 is a new member of the hypoxic gene family. Gene disruption showed that SUT1 is not essential for aerobic or anaerobic yeast growth. PMID- 7489927 TI - Pancreatic inflammatory disease. PMID- 7489926 TI - The management of chronic hepatitis C virus infection. PMID- 7489928 TI - Reflux related symptoms in patients with normal oesophageal exposure to acid. AB - Several studies, using pH monitoring with event markers, have identified patients with normal oesophageal exposure to acid despite an apparent relation between symptoms and reflux episodes. In this series of 771 consecutive patients referred for 24 hour oesophageal pH monitoring, a probability calculation was used to evaluate the relation between symptoms and reflux episodes. Oesophageal exposure to acid was normal in 462 of 771 recordings (59.9%); despite this, 70.8% (327 of 462) of these patients used at least once the event marker. In 96 patients (12.5% of total patients) with normal oesophageal exposure to acid, there was a statistically significant association between symptoms and reflux episodes. The symptom cluster of such patients was similar to that usually seen in patients with gastro-oesophageal reflux disease, but symptoms like belching, bloating, and nausea were common thus overlapping with the symptom pattern of functional dyspepsia. In these patients both the duration and the minimum pH of reflux episodes (either symptom related or asymptomatic) were significantly shorter and higher, respectively, when compared with those of patients with gastro oesophageal reflux disease. These results are consistent with the idea that oesophageal hypersensitivity to acid is the underlying pathophysiological feature of this syndrome. PMID- 7489929 TI - Alkaline gastro-oesophageal reflux: dual probe pH monitoring. AB - Although the aetiology of Barrett's oesophagus or columnar line oesophagus (CLO), remains unknown, bile reflux has been implicated as a factor in its pathogenesis. This study aimed to detect alkaline reflux in gastro-oesophageal reflux patients using dual probe pH monitoring. Thirty patients with histologically diagnosed CLO, 15 age and sex matched patients with oesophagitis (grade 1-3), and 15 healthy volunteers were studied by dual probe, 18 hour pH monitoring and analysis of the bile acid content of oesophageal refluxate. Total acid exposure and acid exposure in the upright and supine postures were greater in CLO subjects than in oesophagitis patients and controls. Furthermore, the number of reflux episodes lasting more than five minutes and the duration of the longest reflux episode were significantly greater in the CLO subjects than the oesophagitis and control subjects. Nine subjects with CLO and oesophagitis, however, were not identified as refluxers, although six had a bile acid concentration in their oesophageal aspirate higher than the 95th centile value of the controls. There was no correlation between the oesophageal pH and the bile acid contents of refluxate. It is concluded that dual probe pH monitoring is not useful in detecting alkaline refluxers. pH monitoring, although the only subjective test available to identify acid refluxers, is not a sufficiently sensitive test with which to define alkaline reflux. PMID- 7489930 TI - Lymphocytic gastritis and Helicobacter pylori infection in gastric lymphoma. AB - Lymphocytic gastritis and primary gastric lymphoma are rare conditions with unknown aetiology. It has recently been suggested that Helicobacter pylori has a role in the pathogenesis of both of them. The occurrence of lymphocytic gastritis and H pylori was studied in a series of patients with primary gastric lymphoma. The cases of primary gastric lymphomas (n = 35) diagnosed in years 1970-1993 were identified. The specimens of 22 cases contained gastric mucosa sufficiently so that the number of intra-epithelial lymphocytes, severity of gastritis, and occurrence of H pylori could be studied. Lymphocytic gastritis was detected in seven of 22 patients (32%), and in most cases both in antral and body mucosa. Atrophy of the body glands was significantly more severe in lymphocytic gastritis patients. H pylori was detected in 13 of all 22 patients (59%); two of seven lymphocytic gastritis patients (29%), and 11 of 15 (73%) of patients without lymphocytic gastritis were H pylori positive. Patients with gastric lymphoma have significantly increased prevalence of lymphocytic gastritis. Rarity of H pylori in these patients might be connected with atrophic changes in body mucosa. Further studies are needed to show the significance of lymphocytic gastritis as a precursor of gastric lymphoma. PMID- 7489934 TI - The caecocolonic junction in humans has a sphincteric anatomy and function. AB - Sphincteric anatomy and function are present at the caecocolonic junction in several mammals. In humans, radiologists and endoscopists have respectively reported a circumferential contraction and a prominent ileocaecal fold at the border area between the caecum and the ascending colon. Anatomical findings on necropsy material failed to confirm its presence. Microscopic studies on surgical specimens showed the existence of muscular and innervational patterns different from those of adjacent areas. The aim of this work was to confirm the existence of a specialised fold at the caecocolonic junction in humans and to ascertain its role by carrying out a study of functional anatomy. Pancolonoscopies were performed on 100 patients and ileocaecal fold behaviour was observed before and after mechanical stimulation. Isolated ileocaecocolonic regions, surgically obtained, were filled with a fixative solution to study their macro and microscopic morphology after stimulation. Endoscopically, the ileocaecal fold was semilunar or circular in shape and spontaneous or evoked spasms occurred in 52 patients. A prominent circular fold could be seen in surgical specimens after stimulation. The entire muscle coat deeply penetrated this fold, showing the features characteristic of the ileocaecal junction. In particular, the inner portion of the circular muscle showed a peculiar arrangement and was thicker than elsewhere. These results show that in humans the caecocolonic junction is provided with a sphincter morphology and function. Little is known about its physiological relevance in ileal flow accommodation and caecal filling and emptying but it should not be underestimated with regard to some colonic motility disorders. PMID- 7489933 TI - Antral motility in patients with cirrhosis with or without gastric antral vascular ectasia. AB - Gastric motility has not been extensively studied in patients with cirrhosis and gastric antral vascular ectasia (GAVE) may be associated with antropyloric dysfunction. This study therefore looked at antral motility using ultrasound in patients with alcoholic cirrhosis with or without GAVE. Twenty six patients were included: 10 patients with cirrhosis without GAVE, eight patients with cirrhosis and GAVE, and eight controls without liver disease. Measurement of antral area and antral contractions (amplitude and frequency) was performed for three hours after ingestion of a standardised solid-liquid meal. Antral area half time (mean (SD)) was not significantly increased in patients with cirrhosis without GAVE (84 (42) min), but increased by 120% (123 (43) min; p < 0.01) in patients with GAVE compared with controls (56 (26) min). GAVE patients exhibited the same frequency and amplitude of antral contractions at each time point as controls and had the same tendency to increase these values over time although this was attenuated in the late postprandial phase. In contrast, cirrhotic patients without GAVE exhibited a significantly higher frequency and amplitude of antral contractions during the initial postprandial phase but showed no change in either frequency or amplitude over time. In conclusion, in cirrhosis there is an abnormal antral motor response to a meal, which has a different pattern over time in patients with or without GAVE. PMID- 7489931 TI - Omeprazole enhances efficacy of triple therapy in eradicating Helicobacter pylori. AB - Triple therapy has been recommended as the most effective treatment for Helicobacter pylori eradication. Despite achieving a comparatively high eradication result, however, around 10% of patients still fail to be cured. Omeprazole can enhance efficacy of single and double antibiotic protocols and is particularly effective when combined with clarithromycin and a nitroimidazole. This study examined the effect of combining triple therapy with omeprazole. A prospective, randomised, unblinded, single centre trial was carried out on consecutive patients with symptoms of dyspepsia and H pylori infection confirmed by rapid urease test, microbiological culture, and histological assessment. Patients were given a five times/day, 12 day course of colloidal bismuth subcitrate chewable tablets (108 mg), tetracycline HCl (250 mg), and metronidazole (200 mg) with either 20 mg omeprazole twice daily (triple therapy+omeprazole) or 40 mg famotidine (triple therapy+famotidine) at night. Compliance and side effects were determined using a standard questionnaire form. One hundred and twenty five of 165 triple therapy+omeprazole patients and 124 of 171 triple therapy+famotidine patients returned for rebiopsy four weeks after completion of treatment. Significantly more triple therapy+omeprazole patients achieved eradication 122 of 125 (97.6%) as assessed by negative urease test, culture, and histological assessment, when compared with 110 of 124 (89%) triple therapy+famotidine patients (p = 0.006; chi 2). There were 30 triple therapy+omeprazole (24%) and 26 triple therapy+famotidine (21%) patients with de novo metronidazole resistant H pylori included in the study. Side effects were mild and infrequent and were comparable in both groups, although pain in duodenal ulcer, gastric ulcer, and oesophagitis patients seemed to subside earlier in those taking omeprazole. Compliance (>95% of drugs taken) was achieved by 98% of patients of both groups. A 12 days regimen of triple therapy with omeprazole is more effective in achieving H pylori eradication than is triple therapy plus famotidine. Use of 20 mg omeprazole twice daily rather than 40 mg famotidine with a 12 day, low dose triple therapy enhances eradication to over 97% whether the H pylori is metronidazole sensitive or resistant. PMID- 7489932 TI - Eradication of Helicobacter pylori restores the inhibitory effect of cholecystokinin on postprandial gastrin release in duodenal ulcer patients. AB - Helicobacter pylori infection may be associated with duodenal ulcer (DU) and accompanied by enhanced gastrin release but the mechanism of this H pylori related hypergastrinaemia in DU patients is unclear. Cholecystokinin (CCK) has been implicated in the feedback control of gastrin release and gastric acid secretion in healthy subjects. This study therefore investigated if CCK participates in the impairment of postprandial gastrin release and gastric secretion in six DU patients. Tests were undertaken with and without elimination of endogenous CCK by loxiglumide, a selective CCK-A receptors antagonist, before and after eradication of H pylori with triple therapy (omeprazole, amoxicyllin, bismuth). In H pylori positive DU patients, the post-prandial decline in pH (with median pH 3.5) was accompanied by a pronounced increment in plasma gastrin but the administration of loxiglumide did not affect significantly this postprandial rise in plasma gastrin and gastric pH profile. After eradication of H pylori, the plasma gastrin concentration was reduced while the median postprandial pH was significantly increased (median pH 4.3). The administration of loxiglumide resulted in significantly greater increase in postprandial plasma gastrin and greater decrease in pH (median pH 3.1) in these patients. This study shows that (a) infection with H pylori is accompanied by an enhanced gastrin release and gastric acidity in DU patients, (b) the failure of loxiglumide to affect plasma gastrin or gastric acid secretion in H pylori infected DU patients could be attributed, at least in part, to the failure of endogenous CCK to control gastrin release and gastric secretion by releasing somatostatin, and (c) the test with loxiglumide may be useful in the identification of patients with impaired feedback control of gastrin release and gastric secretion resulting from infection with H pylori. PMID- 7489937 TI - Risks of gastrointestinal bleeding during secondary prevention of vascular events with aspirin--analysis of gastrointestinal bleeding during the UK-TIA trial. AB - From 1979 to 1985 2435 patients having had transient ischaemic attacks (TIAs) or minor ischaemic strokes, were enrolled in the UK TIA trial and were randomised to receive either aspirin 300 mg, daily or aspirin 1200 mg or placebo. Analysis of reported upper gastrointestinal bleeding events (defined as haematemesis or melaena, or both) showed a risk of bleeding in a dose dependent manner, odds ratios (95% CI) for 300 mg of aspirin = 3.3 (1.2 to 9.0) and for 1200 mg = 6.4 (2.5 to 16.5) and, as would be expected, an increased risk of hospitalisation because of bleeding also in a dose dependent manner, odds ratio = 3.6 (0.7 to 17.2) for 300 mg and 8.7 (2.0 to 37.6) for 1200 mg. Further analysis suggested greater risks of bleeding from duodenal ulcers than gastric ulcers and that bleeding is more likely early in the course of treatment with aspirin used as secondary prevention. There was also an increased risk of lower gastrointestinal bleeding, defined as fresh blood per rectum for both doses of aspirin, odds ratio 1.8 (0.5 to 6.1) for 300 mg of aspirin, and 1.5 (0.4 to 5.3) for 1200 mg of aspirin. PMID- 7489935 TI - Hyperventilation, central autonomic control, and colonic tone in humans. AB - Symptoms attributable to hyperventilation are common among patients with the irritable bowel syndrome (IBS); indeed, some have suggested that hyperventilation may exacerbate the alimentary symptoms of IBS. Hyperventilation changes haemodynamic function through central and peripheral mechanisms; its effects on colonic motor function, however, are unknown. The aim of this study, therefore, was to assess the effects of hyperventilation on colonic tone and motility and on cardiovascular autonomic activity, and to discover if hypocapnia was critical to elicit the response. Phasic and tonic motility of the transverse and sigmoid colon, end tidal PCO2, pulse rate, and beat to beat pulse variability were assessed before, during, and after a five minute period of hypocapnic hyperventilation in 15 healthy volunteers; in seven other subjects, effects of both eucapnic and hypocapnic hyperventilation were evaluated. Hypocapnic but not eucapnic hyperventilation produced an increase in colonic tone and phasic contractility in the transverse and sigmoid regions and an increase in pulse rate and pulse interval variability. The findings are consistent with inhibition of sympathetic innervation to the colon or direct effects of hypocapnia on colonic smooth muscle, or both. These physiological gut responses suggest that some of the changes in colonic function are caused by altered brain or autonomic control mechanisms. PMID- 7489936 TI - Quality of life measurement in patients with oesophageal cancer. AB - Quality of life (QOL) measurement may aid decision making in the treatment of patients with oesophageal cancer but must be clinically valid to be useful. This study considered if the European Organisation for Research and Treatment of Cancer QOL questionnaire, the QLQ-C30, showed differing results in two clinically distinct groups of patients with oesophageal cancer and also investigated the correlation between dysphagia grade and various scales of QOL. Patients treated by oesophagectomy reported significantly better physical, emotional, cognitive, and global health scores than those in the palliative treatment group. Patients who received palliative treatment had significantly worse pain, fatigue, appetite loss, constipation, and dysphagia. The correlations between dysphagia grade and each of the QOL scales and items in both groups of patients were poor. This questionnaire differentiates clearly between the two clinically distinct groups of patients, but to be an entirely appropriate indicator of QOL in patients with oesophageal cancer, an additional specific oesophageal module including a dysphagia scale is required. PMID- 7489938 TI - Attachment of Giardia lamblia trophozoites to a cultured human intestinal cell line. AB - Attachment of Giardia lamblia trophozoites to enterocytes is essential for colonisation of the small intestine and is considered a prerequisite for giardia induced enterocyte damage. The precise mechanisms involved are still being debated and some earlier work has been performed in models of uncertain biological relevance. In this study, co-incubation of giardia with enterocyte like differentiated Caco-2 cells was used as a model to study the influence of physical and chemical factors on attachment. Giardia attachment was maximal between one and eight hours and stable over pH 7.2-8.2 but it was reduced by acidification. Attachment was dependent on temperature and was maximal at 37 degrees and virtually abolished at 4 degrees C. It was reduced compared with controls (p < 0.05) by EDTA 2.5 mM (mean (SEM) 32 (4)%), colchicine 12.5 microM (35 (5)%), mebendazole 10 micrograms/ml (30 (3)%), and cytochalasin B 1 microgram/ml (34 (3)%). Giardia attachment was also diminished by preincubation with mannose 50 mM or mannose-6-phosphate 35 mM (21 (4); 17 (5)%) or by preincubating Caco-2 cells with concanavalin A 100 micrograms/ml (19 (2)%). Enhanced binding was not evident after trypsinisation of trophozoites. Scanning electron microscopy showed that giardia seemed to attach to the Caco-2 monolayer predominantly by its ventral surface but dorsal orientation was also observed. No difference in attachment was observed between three different giardia isolates or a parent isolate and its clone. Attachment of giardia to Caco-2 cells is primarily by cytoskeletal mechanisms, inhibitable by interference with contractile filaments and microtubules, while attachment by mannose binding lectin also seems to mediate binding. PMID- 7489939 TI - Immunohistochemical findings in jejunal specimens from patients with IgA deficiency. AB - Jejunal biopsy specimens from 25 patients with IgA deficiency (IgAd) were studied immunohistochemically to find markers of inflammation. Five of the 25 patients had coeliac disease (CD): they were on a gluten free diet and had normal jejunal morphology. Only two of 15 specimens from control subjects had CD25+ cells in the surface epithelium, while this was seen in 19 out of 20 specimens from IgAd patients (p < 0.0001). A significant increase of CD25+ cells was also noted in the lamina propria of IgAd patients. The median percentage of crypt cells in mitosis (Ki67+ cells) was higher in the specimens from IgAd patients (26%) than in those from controls (13%, p < 0.001). The densities of gamma delta T cell receptor positive cells in the surface epithelium and lamina propria did not differ in the specimens from IgAd patients and those of controls nor was the expression of HLA class II antigens augmented in the surface epithelium. These findings were similar for the IgAd patients whether or not the patient had DQB 0201 allele, a genetic marker which is strongly associated with CD. The inadequacy of the local immunoglobulins in patients with IgAd may lead to increased T cell activation, which is accompanied by the appearance of intraepithelial CD25+ cells and with an increase in the mitotic rate in the crypts. PMID- 7489940 TI - Loss of CD4 T lymphocytes in patients infected with human immunodeficiency virus type 1 is more pronounced in the duodenal mucosa than in the peripheral blood. Berlin Diarrhea/Wasting Syndrome Study Group. AB - Although changes in T lymphocyte subset distribution in the peripheral blood of patients infected with human immunodeficiency virus (HIV) are well defined it is not known whether these changes reflect changes in lymphoid compartments clearly involved in HIV related disease like the intestinal mucosa. This study analysed lymphocytes isolated simultaneously from the peripheral blood and duodenal biopsy specimens by three colour flow cytometry in eight asymptomatic HIV infected patients, 26 AIDS patients, and 23 controls. The proportion of CD4, CD8, CD4-CD8 , or gamma delta T cells did not correlate between circulating and duodenal T cells. CD4 T cells were reduced in the peripheral blood (7.5% (25th-75th percentile, 2-16%) v 52% (41-63%), p < 0.0005) and even more reduced in the duodenum (1% (1-2%) v 36% (23-57%), p < 0.0005) of AIDS patients compared with controls. Patients with asymptomatic HIV infection had intermediate CD4 T cells in the peripheral blood (24% (22-35%); p < 0.002 v controls; p < 0.01 v AIDS) but like AIDS patients very low CD4 T cells in the duodenum (3% (1-6%); p < 0.002 v controls). The ratio of duodenal to circulating CD4+ T cells was significantly reduced to 0.2 (0-1) in AIDS patients (p < 0.001) and even to 0.1 (0.04-0.5) in asymptomatic HIV infected patients (p < 0.002) compared with 0.72 (0.44-0.95) in controls. These findings show an early and preferential loss of duodenal CD4 T cells in HIV infection. Immunological abnormalities in HIV infection are distinct between lymphoid compartments, and profound immunodeficiency may occur in the intestinal immune system although circulating T cells are largely preserved. PMID- 7489941 TI - Colitis and colonic mucosal barrier dysfunction. AB - Trauma, infection, neoplasia, and inflammation can all disrupt the intact intestinal mucosal barrier to intraluminal bacteria and bacterial antigens. This study investigated the relation between colonic inflammation and colonic mucosal barrier function in three experimental models of colitis. There were significantly increased systemic endotoxin concentrations in rats with acetic acid (7.5 (1.7-119.5) pg/ml), ethanol (13.7 (0-111.2) pg/ml), and hapten induced (14.4 (5-31.1) pg/ml) colitis compared with saline controls (3.3 (0-13.7) pg/ml). Data expressed as median (range). There were significant correlations between the systemic endotoxin concentration and both the severity of colitis and of illness in acetic acid induced colitis. A significant increase in colonic permeability to 14C-polyethylene glycol was shown in rats with acetic acid (3.42 (1.36-5.63)%) and hapten induced colitis (2.86 (1.03-8.10)%) compared with saline controls (1.20 (0.67-1.36)%). Data expressed as median (range) of percentage of the intracolonic bolus excreted in urine. There was a significant positive correlation between the severity of colitis and % colonic permeability to 14C polyethylene glycol. This and other studies provide evidence that mucosal barrier dysfunction is a feature of colitis irrespective of aetiology or species. Such barrier dysfunction may be responsible for the systemic inflammatory response and complications seen in patients with inflammatory bowel disease. PMID- 7489942 TI - Interleukin 8 secretion by colonic crypt cells in vitro: response to injury suppressed by butyrate and enhanced in inflammatory bowel disease. AB - Epithelia from several sites exhibit inducible secretion of interleukin 8 (IL-8). This study aimed to characterise secretion of IL-8 by colonic epithelial cells in vitro. Colonic crypt cells were isolated enzymatically from resected colon and the IL-8 content of culture supernates was measured by ELISA. The rate of secretion of IL-8 accelerated and levels of IL-8 transcripts increased appreciably during culture. Exposure to tumour necrosis factor alpha (TNF alpha) failed to increase secretion further. Secretion was not induced by the enzymatic digestion or by serum used in the culture medium but was significantly inhibited by butyrate, by a mean of 23%. Control experiments indicated that colonic crypt cells were the likely source. The secretion of IL-8 over 24 hours by cells from uninflamed mucosa of patients with ulcerative colitis or Crohn's disease was more than twofold that from normal cells, while that from cancer bearing colons was normal. TNF alpha (10 mM) significantly suppressed IL-8 secretion only in the ulcerative colitis group and the change was different to those in the normal (p = 0.007) and Crohn's disease groups (p = 0.012). Cells from inflamed areas secreted more IL-8 than those from autologous uninflamed areas (p = 0.009) but responses to modulating factors were no different. The induction of IL-8 secretion by colonic crypt cells in vitro is probably a response to injury associated with isolation and culture. It is suppressed by butyrate and increased in inflammatory bowel disease independently of the presence of mucosal inflammation. Whether epithelial derived IL-8 plays a part in the pathogenesis of inflammatory bowel disease is not yet clear. PMID- 7489943 TI - Corticotropin releasing hormone in colonic mucosa in patients with ulcerative colitis. AB - Corticotropin releasing hormone (CRH) is a key hormone in integrated response to stress, acting as the major regulator of the hypothalamic-pituitary-adrenal axis. Recently, local production of CRH has been detected in normal human colonic enterochromaffin cells. CRH is locally secreted in granulomatous and arthritic tissues in rats and humans, where it seems to act as a local proinflammatory agent. To find out if CRH is present in colonic tissues of patients with ulcerative colitis, this study examined the expression of this peptide in the large bowel of patients with ulcerative colitis. Colonic tissues of patients with ulcerative colitis obtained by endoscopic biopsy were immunostained with anti-CRH antibody. CRH messenger (m) RNA was also examined in biopsy specimens of ulcerative colitis by the reverse transcribed polymerase chain reaction method and by in situ hybridisation. Considerably enhanced expression of immunoreactive CRH was found in mucosal inflammatory cells. Intense staining with anti-CRH antibody was also shown in mucosal macrophages. CRH mRNA was expressed in mucosal epithelial cells. The expression of immunoreactive CRH in colonic mucosal epithelial cells of ulcerative colitis slightly increased, but not significantly, compared with normal colonic mucosal epithelial cells. These results suggest that CRH may play a part in the modulation of intestinal immune and inflammatory system, and as a modulator in the pathogenesis of ulcerative colitis. PMID- 7489944 TI - The perfect pelvic pouch--what makes the difference? AB - The aim of this study was to determine what factors are important for the achievement of perfect anal continence after restorative proctocolectomy. One hundred patients underwent paired studies of anorectal physiology before and one year after restorative proctocolectomy with pelvic ileal reservoir (11 S, 25 J, 64 W) with stapled ileoanal anastomosis, without mucosectomy. Fifty seven patients attained perfect anal continence and were able to discriminate flatus from faeces with such confidence that they were able to release flatus safely without fear of faecal soiling. The remaining 43 patients experienced minor problems in this regard. Four factors were found to correlate significantly with a perfect functional result (median, perfect v imperfect): maximum resting anal pressure (72 v 57 cm H2O, p < 0.02), the sensory threshold in the upper and mid anal canal (7.3 v 8.6 and 5.3 v 7.0 mA, p < 0.05 and p < 0.02), compliance of the ileal reservoir (12.4 v 7.6 ml/cm H2O, p < 0.01), and the presence of a pouch anal inhibitory reflex (56 of 57 patients v 29 of 43 patients, p < 0.01). The quality of anal continence depends on several factors: a complaint ileal reservoir, a strong sensitive anal sphincter, and normal reflex coordination of the activities of the reservoir and the sphincter. Excellent pouch-anal coordination is obtainable irrespective of the design of the reservoir, provided that these criteria are satisfied. PMID- 7489945 TI - Comparison of 99m technetium hexamethylpropylene-amine oxime labelled leucocyte with 111-indium tropolonate labelled granulocyte scanning and ultrasound in the diagnosis of intra-abdominal abscess. AB - Fifty patients with suspected intra-abdominal abscess were investigated prospectively with ultrasound and with 99mTc-hexamethylpropylene-amine oxime (HMPAO) isotope labelled mixed leucocytes, using 111-In tropolonate granulocyte scanning as the reference standard. Twenty five patients had inflammatory bowel disease (three were postoperative): 21 of these had Crohn's disease and four had ulcerative colitis. The remainder comprised nine with postoperative fever and 16 with fever and abdominal pain. An abscess was diagnosed when focal activity on serial 111-In tropolonate and 99m-Tc-HMPOA images at one, three, and 24 hours resulted in activity at least equal to liver activity at 24 hours. Thirteen abscesses were diagnosed using each type of white cell scanning, resulting in 100% sensitivity for 99m-Tc-HMPAO compared with 111-In tropolonate. Bowel inflammation was easily distinguished from abscess on serial images. Eight of these 13 abscesses were detected by ultrasound. Altogether 17 abscesses were found. Ultrasound detected 12, including four liver abscesses which were not purulent and had not been detected by white cell scanning. Ultrasound had a sensitivity of 71% (12 of 17) and a specificity of 87% (33 of 38) using all confirmed abscesses as the reference standard. White cell scanning showed a sensitivity of 76% (13 of 17: as a result of the four non-purulent liver abscesses) and a specificity of 100%. 99m-Tc-HMPAO scanning is as accurate as 111 In tropolonate scanning, and has several advantages including simplicity, availability, superior image quality, and reduced radiation dose. Both methods are more sensitive and specific than ultrasound for intra-abdominal abscess detection but ultrasound is advisable if a neutrophil infiltrate is not suspected. PMID- 7489946 TI - Drug induced acute pancreatitis: incidence and severity. AB - To determine the incidence and severity of drug induced acute pancreatitis, data from 45 German centres of gastroenterology were evaluated. Among 1613 patients treated for acute pancreatitis in 1993, drug induced acute pancreatitis was diagnosed in 22 patients (incidence 1.4%). Drugs held responsible were azathioprine, mesalazine/sulfasalazine, 2',3'-dideoxyinosine (ddI), oestrogens, frusemide, hydrochlorothiazide, and rifampicin. Pancreatic necrosis not exceeding 33% of the organ was found on ultrasonography or computed tomography, or both, in three patients (14%). Pancreatic pseudocysts did not occur. A decrease of arterial PO2 reflecting respiratory insufficiency, and an increase of serum creatinine, reflecting renal insufficiency as complications of acute pancreatitis were seen in two (9%) and four (18%) patients, respectively. Artificial ventilation was not needed, and dialysis was necessary in only one (5%) case. Two patients (9%) died of AIDS and tuberculosis, respectively; pancreatitis did not seem to have contributed materially to their death. In conclusion, drugs rarely cause acute pancreatitis, and drug induced acute pancreatitis usually runs a benign course. PMID- 7489948 TI - Is day case liver biopsy underutilised? AB - Day case liver biopsies are rarely performed nationally but have been routine practice in selected patients in our hospital since 1989. We have audited our experience of this procedure to compare its safety, and efficacy with inpatient biopsy and assess patient acceptability. Audit data were collected retrospectively on liver biopsies performed at a teaching hospital over 42 months. Acceptability of day case biopsy was assessed by a questionnaire. A total of 182 of 546 biopsies were day cases (33%). The specimen quality was similar in both groups. The overall complication rate did not significantly differ between the two groups (2.7% day case v 3.3% inpatients). There were no deaths or episodes of haemorrhage in the day cases but one patient developed a pneumothorax. Some 91% of those who had a day case biopsy were satisfied with the procedure. Day case liver biopsy is safe, effective, and acceptable in selected patients. PMID- 7489947 TI - Clinical relevance of mutations in the precore genome of the hepatitis B virus. AB - A stop codon in the precore genome of the hepatitis B virus (HBV) in anti-HBe positive HBV carriers may be associated with a more progressive form of HBV infection. Earlier studies, however, were mainly performed in patients from the Mediterranean area who had severe infection. The aim of this study was to evaluate the prevalence of precore mutants in an unselected population living in northern Europe. Twenty of 42 of these patients are infected predominantly with a virus strain, which has the typical stop codon in the precore genome, characterised by a mutation at base 83. In six patients there was an additional G to A mutation at base 86 of the precore genome. Statistical analysis showed no difference between the patients with or without a stop codon in the precore genome. When patients with a double mutation at base 83 and 86 of the precore genome were compared with the other anti-HBe positive HBV carriers, however, the corresponding clinical data were worse. Therefore we suggest, that it is not the stop codon in the precore gene itself, but the occurrence of a double mutation at bases 83 and 86, which is associated with a more severe course of disease in anti HBe positive HBV carriers. PMID- 7489949 TI - Selective preoperative endoscopic retrograde cholangiography with sphincterotomy avoids bile duct exploration during laparoscopic cholecystectomy. AB - A policy of preoperative endoscopic retrograde cholangiography (ERC) for suspected bile duct stones was used in 1507 patients considered for laparoscopic cholecystectomy in three district general hospitals. Altogether 306 patients underwent ERC, and bile duct cannulation was achieved in 276 (90%). Bile ducts were cleared by endoscopic sphincterotomy in 128 of 161 patients (79%) with proven duct stones. Laparoscopic cholecystectomy was completed in 1396 patients. Ten laparotomies were necessary for complications of laparoscopic cholecystectomy. The complication rate for endoscopic sphincterotomy/laparoscopic cholecystectomy was 2.7%, with no mortality. Overall, a combined endoscopic/laparoscopic approach succeeded in 1386 patients (92%). Fourteen patients (1%) had retained stones during a median of 14 months (range 1-42) follow up, all of which were removed by ERC/endoscopic sphincterotomy. If a policy of selective ERC before laparoscopic cholecystectomy is used for all patients with symptomatic gall stones, most will avoid an open operation and laparoscopic exploration of the bile duct is not necessary. PMID- 7489950 TI - Fetal mortality associated with cholestasis of pregnancy and the potential benefit of therapy with ursodeoxycholic acid. AB - Cholestasis of pregnancy is associated with increased fetal morbidity and mortality and should be treated actively. The significance attached to pruritus in pregnancy is often minimal, but it is a cardinal symptom of cholestasis of pregnancy, which may have no other clinical features. Eight women with previous cholestasis of pregnancy were referred to The Liver Unit within a 12 month period for advice concerning future pregnancies. Thirteen pregnancies had been affected by cholestasis of pregnancy and 12 had been treated expectantly with resultant perinatal morbidity or mortality in 11 (one normal delivery), including; eight stillbirths, two premature deliveries with fetal distress (one died in perinatal period), and an emergency caesarean section for fetal distress. The other pregnancy was treated actively and delivery was uncomplicated. Subsequently, three of these cases with recurrent cholestasis of pregnancy were referred while pregnant. In each, cholestasis developed with severe pruritus, gross increase of serum bile acids, and deranged liver tests. Each was treated with the choleretic agent ursodeoxycholic acid, with rapid clinical improvement and resolution of deranged biochemistry. In conclusion, cholestasis of pregnancy continues to be treated expectantly despite its association with increased morbidity and mortality and evidence suggesting improved prognosis with active treatment and the potential of reducing the associated perinatal mortality. In an uncontrolled series of three patients with cholestasis of pregnancy, ursodeoxycholic acid seemed to provide safe and effective therapy. PMID- 7489951 TI - 'Sticky' neutrophils, pathergic arthritis, and response to heparin in pyoderma gangrenosum complicating ulcerative colitis. AB - Pyoderma gangrenosum is strongly associated with inflammatory bowel disease and exhibits pathergy, occurring at sites of previous minor trauma. A patient is presented with a 21 year history of extensive ulcerative colitis, who developed pyoderma gangrenosum and arthralgia while receiving high dose corticosteroids for active ulcerative colitis. The arthralgia exhibited pathergy affecting particularly the left temporomandibular joint, which was stressed by an asymmetric bite, and the left elbow, which had been fractured many years previously. This prompted the hypothesis that neutrophils in this condition may be marginated, as a result of increased stickiness of either the neutrophil or the vascular endothelium. The introduction of heparin therapy was associated with rapid resolution of the arthralgia, pyoderma gangrenosum, and ulcerative colitis. PMID- 7489952 TI - Hydatid disease. PMID- 7489953 TI - Intestinal permeability. PMID- 7489954 TI - Gastric mucus viscosity and Helicobacter pylori. PMID- 7489955 TI - Copper associated childhood cirrhosis. PMID- 7489956 TI - Eradication rate of Helicobacter pylori. PMID- 7489957 TI - Eradication rate of Helicobcter pylori. PMID- 7489958 TI - The haemorrhagic effect of low molecular weight heparins, dermatan sulphate and hirudin. AB - In a randomized, blind study the primary effect on haemostasis after intravenous administration of dermatan sulphate (DS), recombinant hirudin (r-hirudin) and four commercial low molecular weight heparins (LMWHs) (nadroparine, enoxaparin, dalteparin and tinzaparin) was investigated in rats and compared with saline (control). The tail bleeding time, the bleeding from the gastric mucosa [the mucosal bleeding time (min) and the mucosal bleeding (microliter)] as well as changes in activated partial thromboplastin time, antifactor IIa and Xa activities were investigated. DS and r-hirudin were investigated in a dose potentially suitable in thomboprophylaxis and the LMWHs in doses recommended by the manufacturers for thromboprophylaxis, adjusted to body weight. All substances significantly prolonged the mucosal bleeding time. Dalteparin, tinzaparin, DS and r-hirudin increased the mucosal bleeding when compared with controls whereas nadroparine and enoxaparin did not. The effect of r-hirudin was also significantly more pronounced compared with other treatments. Moreover, r-hirudin prolonged the tail bleeding time significantly whereas the other substances did not. The antifactor Xa activity in plasma correlated well with the given dose of the LMWHs (rs = 0.7). However, the monitored bleeding parameters in the LMWH groups did not correlate with the plasma activities of antifactor IIa or Xa. The results indicate that the tested LMWHs are not equipotent in their effect on haemostasis in this model and that antifactor IIa or Xa activities do not directly correlate with their effect on haemostasis although increased haemorrhage was observed in the LMWHs with lower antifactor Xa/antifactor IIa ratios.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7489959 TI - Preventive effect of long-term aerobic exercise on thrombus formation in rat cerebral vessels. AB - The antithrombotic effect of long-term aerobic exercise was studied in rat cerebral blood vessels using an argon (Ar) laser-induced thrombosis technique. Rats were subjected to treadmill exercise at 70% VO2max for 2 and 4 months. The number of laser irradiations required to form an occlusive thrombus in the exercise group was significantly increased compared to that of the control group after 2- and 4-month training periods. Fibrinolytic activity increased just after exercise but there was no significant difference between the exercise and the control group in the resting state after a period of exercising conditioning. The results demonstrated that long-term aerobic exercise decreased the thrombotic tendency in rat cerebral vessels and suggested that such exercise might have a protective effect on these blood vessels. PMID- 7489960 TI - Disseminated intravascular coagulation in endemic hepatosplenic schistosomiasis. AB - Thrombin and plasmin generation were assessed in patients with endemic hepatosplenic schistomiasis (15 hepatosplenomegalic, 15 splenomegalic, 15 with advanced hepatic fibrosis and ascites and 15 hepatosplenic patients with hematemesis). Prolongation of prothrombin time, partial thromboplastin time and thrombin time, thrombocytopenia, hypofibrinogenemia, a decrease in antithrombin III and protein C and S levels and elevation in fibrinopeptide A, D-dimer and thrombin-antithrombin complex levels were detected in all groups. The deficit in hemostatic parameters was more pronounced with the advancement of the disease and was maximal in the hematemesis group. Our data demonstrate an increase in both thrombin and plasmin generation and indicate that low grade disseminated intravascular coagulation may occur in association with endemic Egyptian hepatosplenic schistosomiasis even in the steady state without overt bleeding. PMID- 7489961 TI - Efficacy and safety of a monoclonal purified factor VIII concentrate: 5-year follow-up in previously treated HIV-negative haemophiliacs. AB - The efficacy and safety of a monoclonal purified factor VIII concentrate (Hemofil M) were assessed in a historically controlled study in 22 HIV-negative patients with haemophilia A, previously treated with various concentrates. Data from 5 years of follow-up were compared with those from the preceding 6 months. Factor VIII consumption remained unchanged. A temporary increase in the number of reported bleedings was attributed to more frequent follow-up visits in the first year. Allergic reactions, inhibitor formation and HIV infection were not seen. Liver function parameters fluctuated in individual patients, and were not related to the ultrapure concentrate used. No clinical evidence of liver insufficiency was seen. The number of CD4-positive lymphocytes was stable, while platelet numbers showed a remarkable increase. We conclude that in previously treated HIV negative haemophiliacs, treatment with a monoclonal purified factor VIII concentrate is efficacious and safe with regard to HIV transmission, allergic reactions, induction of inhibitors, and deterioration of immune abnormalities. PMID- 7489962 TI - Haemostatic variables in arterial hypertension. AB - In 22 untreated patients with uncomplicated essential hypertension and in 10 normotensive subjects the plasma levels of thrombomodulin (TM), beta thromboglobulin (beta-TG), D-dimer (DD), tissue-type plasminogen activator (t-PA) and plasminogen activator-inhibitor (PAI-1) were evaluated. The observed values show no significant difference in plasma TM, plasma and urine beta-TG concentration and plasma DD among hypertensive patients and controls. On the other hand, the levels of t-PA and PAI-1 in hypertensive patients were significantly higher than the values detected in normotensive control subjects. These data seem to indicate that, at initial stages of essential hypertension, the t-PA and PAI-1 levels increase. PMID- 7489963 TI - Effects of desmopressin on hemostasis in patients with liver cirrhosis. AB - The aim of this double-blind, placebo-controlled crossover study was to investigate the effect of 1-deamino-8-D-arginine vasopressin (dDAVP) on hemostasis in patients with chronic liver disease. Nine consecutive patients with biopsy-proven liver cirrhosis and related coagulation abnormalities received in a random order dDAVP, 0.5 microgram/kg, or saline intravenously. Blood samples were taken before dDAVP infusion and 30, 60 and 180 min after its end. dDAVP infusion induced a statistically significant shortening of the bleeding time from 9 min (range 6.5-15.5) to 6 min (range 4.5-9.5) at 1 h after the infusion. The activated partial thromboplastin time was significantly shortened at 30 and 60 min after dDAVP infusion. Plasma levels of factor VIII, XI and XII coagulant activities were significantly increased at all sampling times after dDAVP infusion. The maximum increase over basal values in plasma levels of factor VIII, XI and XII was 63, 22 and 40%, respectively. dDAVP did not induce any significant changes of prothrombin time, thrombin clotting time, fibrinogen, plasma levels of factor II, V, VII, IX, X, factor XII antigen, protein C (activity and antigen), antithrombin III, plasminogen and alpha 2-antiplasmin. Placebo infusion did not produce any significant changes in the evaluated parameters. We conclude that dDAVP can positively influence the hemostatic system in patients with liver cirrhosis. The clinical relevance of this hemostatic improvement deserves further evaluation. PMID- 7489964 TI - Plasminogen activators and plasminogen activator inhibitor before and after surgery in patients with and without gastric malignancy. AB - Pre- and postoperative plasma levels of tissue-type plasminogen activator (t-PA), single-chain urinary plasminogen activator, and plasminogen activator inhibitor 1 (PAI-1) were measured in 40 patients undergoing gastric surgery in order to compare patients operated for benign (n = 21) and malignant (n = 19) disease. On the 1st postoperative day, the PAI-1 activity was significantly increased and the t-PA activity significantly decreased in the malignant group, whereas only insignificant changes were seen in the benign group. In contrast, the t-PA activity was significantly increased 1 week after surgery in patients without malignancy. In both groups, the single-chain urinary plasminogen activator activity decreased on the 1st postoperative day, but was significantly increased 1 month after operation. Thus, the two groups reacted differently, but it remains to be determined whether this bears any relationship to the well-known high risk of postoperative venous thromboembolism in cancer patients. PMID- 7489965 TI - Recombinant interferon-alfa therapy in children with chronic hepatitis C. AB - The efficacy of recombinant interferon-alfa therapy in children with chronic hepatitis C has been evaluated in a randomized, controlled pilot study including 27 patients, aged 2 to 14 years, without underlying systemic diseases. On entry, all patients had abnormal alanine transaminase (ALT) levels, 22 were hepatitis C virus (HCV) RNA positive, 19 had mild chronic active hepatitis, and 8 had chronic persistent hepatitis on liver biopsy. Fourteen children received 5 MU/m2 of recombinant interferon-alfa2b thrice weekly for 4 months. If at this time ALT had been reduced to at least 50% the baseline level, treatment was continued up to 12 months. The other 13 children remained untreated. The whole follow-up period lasted 24 months. Interferon was stopped at 4 months in 4 children because of an ALT increase (2 cases), unchanged ALT and febrile convulsions (1 case), and slight ALT decrease (1 case). This latter patient, however, had normal ALT at 6 months and throughout further follow-up, and cleared HCV RNA, thus behaving as a sustained responder. All 10 children treated for 12 months had normal levels of ALT, and 9 were HCV RNA negative at the end of treatment. Of the 9 children who could be followed to 24 months, 4 relapsed soon after therapy withdrawal and 5 maintained a sustained biochemical and virologic response. Overall, 6 (43%) of 14 treated children had a sustained ALT normalization associated with HCV RNA clearance as compared with only 1 (7.5%) untreated child who had a sustained ALT normalization but did not clear HCV RNA.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7489966 TI - Fulminant hepatitis associated with hepatitis B virus e antigen-negative infection: importance of host factors. AB - The precore stop-codon variant of hepatitis B virus (HBV) has been implicated in fulminant hepatitis. The precore/core regions of such variants from two sets of patients with interpartner transmission resulting in fulminant hepatitis in the contact, were sequenced to establish whether further sequence variations in the core region are specifically associated with the fulminant disease. In both sets of patients, there was sequence diversity of the precore/core region from the wild type, leading to numerous amino acid substitutions in the core region. Between the infecting source and the contact, there was only one amino acid change in one set of patients and none in the other. In addition, in the second set of patients, serum samples from four different time points were investigated. Sequence data showed no variation in each patient at the nucleotide level in the core region, even in the case of the source, who was followed for 3 years. In this same pair of subjects, the remainder of the genome was sequenced and was identical at the nucleotide level. Therefore, it appears that, at least in some cases of fulminant hepatitis caused by infection with the precore variant, the nucleotide sequence of the patient with fulminant hepatitis is identical to that observed in the asymptomatic source of infection. These data indicate that the severity and outcome of infection in such cases are unrelated to any additional variation in the entire HBV genome, and that the changed clinical picture is dependent on host factors, possibly the HLA environment.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7489967 TI - Immunoglobulin M antibody to hepatitis C virus core antigen: correlations with viral replication, histological activity, and liver disease outcome. AB - Immunoglobulin M (IgM) antibody to hepatitis C core antigen (anti-HCV-core) was tested by enzyme immunoassay against a synthetic peptide representing amino acids 1 to 62 of the core protein. Of 214 patients with different categories of histological activity, 193 (90%) showed positive results for IgM anti-HCV-core, and 207 (97%) had HCV RNA; most cases (186, 87%) had both markers detectable simultaneously. No differences in the frequency of IgM anti-HCV-core were observed with respect to epidemiological, biochemical, or histological parameters. In 175 interferon alfa (IFN-alpha) recipients, and in 39 untreated controls, pretreatment IgM anti-HCV-core frequencies were similar: 28 of 32 (88%) in sustained responders; 55 of 61 (90%) in responders with relapse; 72 of 82 (88%) in nonresponders; and 38 of 39 (97%) in untreated controls. After IFN-alpha therapy, IgM anti-HCV-core levels became undetectable with significantly greater frequency in sustained responders (P = .014); a similar trend was observed for HCV RNA (P < .0001). IgM anti-HCV-core levels decreased after therapy in responders (P < .001) but increased in nonresponders. Fifty-one cases were longitudinally tested in relation to long-term disease outcome. Both markers remained detectable in most nonresponders with persistent liver disease, in most responders before relapse, and in all but one case at the time of biochemical relapse. IgM anti-HCV-core and HCV RNA became undetectable in most sustained responders, but reappeared despite a long-lasting transaminase normalization, behaving as asymptomatic HCV carriers; the possibility that disease reactivation may take place years afterwards cannot be excluded.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7489968 TI - Hepatitis B virus infection: precore mutants and its relation to viral genotypes and core mutations. AB - The precore-core gene of hepatitis B virus (HBV) was directly sequenced from serum samples of 42 patients with chronic B hepatitis (19 hepatitis B e antigen [HBeAg]+ and 23 anti-HBE+). Viral genotypes were determined by comparison with 11 reference sequences and by restriction analysis. Genotype A was identified in 16 cases, genotype D in 24 cases, and other genotypes in 2 cases. Precore mutations, mainly M1 (stop at codon 28), were differently distributed among the viral genotypes: 3 cases (18.8%) with genotype A and 18 cases (75%) with genotype D. In sequences with precore mutants, the encapsidation signal was more stable (negative stabilization energy) than in sequences without precore mutants. In genotype A, the M1 mutation coexisted with a second mutation (C-->T at position 1858 in codon 15), and both mutations were paired in the secondary structure of the RNA encapsidation signal, which justified the rare presence of precore mutants in this genotype. The analysis showed different distribution of mutations depending on the viral genotype; patients with genotype D were more likely to have persistent HBV infection by selection of precore mutants. Multiple amino acid substitutions were detected in the core region, mainly in two subsequences that have been previously described as epitopes (flanked by codons 11 to 27 and 74 to 83); the presence of these mutations was significantly related to the presence of precore variants which abolished the expression of HBeAg.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7489969 TI - Proteolytic activity of NS3 serine proteinase of hepatitis C virus efficiently expressed in Escherichia coli. AB - The serine proteinase of hepatitis C virus (HCV) non-structural protein NS3 was efficiently expressed in an active form as a fused protein with oligohistidine in Escherichia coli. The recombinant fusion protein was purified to near homogeneity by affinity chromatography on a metal chelation column. Trans-cleavage activity of this protein was investigated by using the substrate NS5 protein expressed in insect cells. The purified serine proteinase trans-cleaved the partially purified NS5 protein. In contrast, the NS3 proteins with mutations at the proposed catalytic site, Ser1165 or His1083, lost the trans-cleavage activity. Analysis of the authentic enzyme and variants with site-directed mutations provides a useful tool for understanding the structure-function relationship of the NS3 serine proteinase. We then developed an in vivo trans-cleavage assay system by coexpression of the NS3 proteinase and the NS5 substrate in E coli, and examined the effect of known inhibitors of serine proteinase. Inhibition of its proteolytic activity by N-p-tosyl-L-lysine chloromethyl ketone (TLCK) was observed, but only at high concentrations. The in vitro and in vivo trans cleavage assays for NS3 serine proteinase will facilitate efficient testing for inhibitors of the replication of HCV and specific treatment for hepatitis C. PMID- 7489970 TI - Evidence of transmission of hepatitis D virus to spouses from sequence analysis of the viral genome. AB - To study sexual transmission of hepatitis D virus (HDV), 52 spouses of 56 index patients were observed and HDV genomes from antibody to HDV (anti-HDV)-positive couples were sequenced. Of the spouses, 11 (21%) were serum HBsAg positive, 3 (27%) of whom were also anti-HDV positive. The HDV sequences between spouses were found to be nearly identical (98% to 98.8%) in the region analyzed (nt 911 to nt 1260). Only one couple showed an identity > 90% with the genotype I HDV strains. The HDV sequences of the remaining two couples showed > 95% identity with each other and > 91% homology with genotype II, but they shared only a 73.1% to 73.7% homology with those of the first couple. The regions corresponding to the autocatalytic cleavage sites, the junction between the middle and the carboxyl terminal one-third domains, and the middle domain of the open reading frame for delta antigen on the antigenomic HDV RNA were more conserved with < 19% divergence among the three couples. Interestingly, there was a 56% divergence in the region corresponding to the carboxyl end of the open reading frame for the large delta antigen on the antigenomic HDV RNA. In summary, this study provides a direct nucleotide evidence of a common source of HDV infection in each couple. Despite divergence in the viral nucleotide sequence, both genotypes I and II were found in Taiwan and were transmitted from patients with a history of prostitute contact to spouses through sexual contact. PMID- 7489971 TI - Parvovirus B19 as a possible causative agent of fulminant liver failure and associated aplastic anemia. AB - We recently observed that more than one third of pediatric patients who presented with non-A, non-B fulminant liver failure (FLF) also developed aplastic anemia (AA) either before or shortly after liver transplantation. Factors involved in the suppression of bone marrow could be the same as those causing hepatic failure. We considered parvovirus B19 a candidate etiologic agent because of the known tropism of B19 for erythroid precursors. Archived liver and serum from six patients undergoing liver transplantation for non-A, non-B, non-C FLF with associated AA were analyzed for the presence of B19 DNA and anti-B19 serology. An age- and gender-matched control group (N = 44) was analyzed in parallel. B19 DNA studies and anti-B19 serology were performed in a blinded fashion. B19 serologies were performed by antibody capture enzyme-linked immunosorbent assay (ELISA). B19 DNA was detected after polymerase chain reaction (PCR) amplification of target B19 DNA sequences in liver and serum. Liver tissue showed evidence of B19 DNA in four of six (66%) patients with FLF and associated AA. Two of 4 patients with cryptogenic FLF but without AA had B19 DNA detected in the liver tissue. Of the 34 remaining controls, only 5 (15%) showed evidence of B19 DNA in liver tissue (66% vs. 15%, P = .016). B19 DNA was not detected in any of the test or control sera. This study provides evidence to support the role of parvovirus B19 in the development of FLF and associated AA. PMID- 7489972 TI - Increased production of nitric oxide by neutrophils and monocytes from cirrhotic patients with ascites and hyperdynamic circulation. AB - An increased release of nitric oxide (NO), a powerful vasodilating agent, has been proposed to play a role in the pathogenesis of vasodilation and hyperdynamic circulation associated with advanced cirrhosis. We evaluated NO synthase (NOS) activity in peripheral leukocytes of 12 cirrhotic patients and 9 healthy subjects together with plasma endotoxin levels and systemic hemodynamic (by a noninvasive echocardiographic method). NOS activity was evaluated by (1) measuring the capacity of isolated polymorphonuclear cells (PMNs) and monocytes to convert [3H]arginine to [3H]citrulline; (2) measuring the ability of neutrophils and monocytes to inhibit thrombin-induced platelet aggregation and to increase guanosine 3'-5'-cyclic monophosphate content in coincubated platelets, an expression of NO release from these cells. Both neutrophils and monocytes from cirrhotic patients produced significantly higher amounts of [3H]citrulline than cells obtained from healthy subjects (P < .001 and P < .02 for neutrophils and monocytes, respectively) and were more effective than control cells in inhibiting platelet aggregation (P < .05 and P < .001, respectively for 2 x 10(6) cells) and in increasing guanosine 3'-5'-cyclic monophosphate content in coincubated platelets (P < .05 and P < .001, respectively). The anti-aggregating activity expressed by leukocytes has a pharmacological profile similar to that described for NO, because it increased after addition of superoxide dismutase, a superoxide anion scavenger, and markedly decreased after inhibition of nitric oxide synthesis with NG-monomethyl-L-arginine (L-NMMA) and NG-nitro-L-arginine-methyl ester (L-NAME). Cirrhotic patients had significantly higher plasma endotoxin levels (P < .001) and cardiac index (P < .01) when compared with controls.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7489973 TI - Management of focal nodular hyperplasia and hepatocellular adenoma in young women: a series of 41 patients with clinical, radiological, and pathological correlations. AB - Preoperative distinction between focal nodular hyperplasia (FNH) that should be managed conservatively and hepatocellular adenoma (HA) that should be resected remains difficult. The result is controversial management of these patients. The aims of this study were to report the value of modern imaging procedures for noninvasive diagnosis of these lesions, to assess the value of intraoperative frozen section studies, and to propose a management strategy in those patients. Forty-one consecutive women with FNH (35 cases) or HA (6 cases) treated at our institution between 1985 and 1992 were studied. New imaging techniques, including enhanced magnetic resonance imaging (MRI) and color Doppler ultrasonography (US), were prospectively appraised in addition to usual techniques. Histological examination of surgical specimens was obtained in all cases. A sixfold increase in the number of patients with FNH was observed during this study, whereas the number of patients with HA did not change. FNHs were incidental US findings in 74% of the cases. The best imaging procedure in the diagnosis of FNH was enhanced MRI with a sensitivity of 70% and a specificity of 98%. Color Doppler US was a useful adjunct. Intraoperative frozen section studies were performed in 16 patients with 19 tumors with a sensitivity of 89% and a specificity of 100%.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7489974 TI - Changes in platelet kinetics after a partial splenic arterial embolization in cirrhotic patients with hypersplenism. AB - We performed a partial splenic arterial embolization in 22 patients with cirrhosis associated with thrombocytopenia and then evaluated the changes in platelet kinetics after undergoing the procedure using 111In-tropolone-labeled platelets. The controls consisted of eight chronic hepatitis patients who showed a normal platelet count and normal spleen size. The mean splenic infarction ratio after the procedure was 54.9%. A platelet kinetics study was performed before and 2 months after the procedure. Before the procedure, the cirrhotic patients showed increases in the splenic volume and the spleen/liver uptake ratio of the 111In labeled platelets on both the third and seventh days, and a decrease in the platelet recovery compared with the controls, which suggested an increased platelet pool in the spleen. In addition, the platelet survival time in cirrhotic patients was shortened, whereas the platelet-associated immunoglobulin G (PA-IgG) was higher than that of the controls, which suggested the involvement of immunologic mechanisms in the thrombocytopenia. With an increase of the platelet count after a partial splenic arterial embolization, the spleen/liver uptake ratio of the 111In-labeled platelets decreased, whereas the platelet recovery increased. Furthermore, the platelet survival time was prolonged, whereas the PA IgG decreased. The platelet count showed a positive correlation with the platelet survival time and a negative correlation with PA-IgG before and after the procedure. These results suggest that a transcatheter splenic arterial embolization not only may reduce the increased platelet pool in the spleen but also may improve the thrombocytopenia induced by immunologic mechanisms in patients with cirrhosis. PMID- 7489975 TI - Paraumbilical vein patency in cirrhosis: effects on hepatic hemodynamics evaluated by Doppler sonography. AB - Doppler sonographic portal vein parameters are used for the noninvasive evaluation of portal hypertension in cirrhosis. The patency of a paraumbilical vein is a rather frequent finding in cirrhosis, which may affect hepatic hemodynamics and function. We evaluated portal and hepatic arterial parameters in 184 cirrhotic patients with and without a patent paraumbilical vein and the relationships with paraumbilical blood flow. A patent paraumbilical vein was found in 33.7% of patients. The prevalence was higher (56.8%) in Child C patients. Portal blood flow velocity (PBV) (10.8 +/- 2.2 vs. 9.8 +/- 2.4 cm/sec; P < .01) and volume (PBF) (995.0 +/- 383.8 vs. 811.6 +/- 318.7 mL/min; P < .001) was significantly higher, and effective portal liver perfusion (PLP) (portal blood flow--paraumbilical blood flow) (621.3 +/- 420.8 vs. 811.6 +/- 318.7 mL/min; P < .001) was significantly lower in patients with a patent paraumbilical vein than in those without. These differences were more evident in Child C patients (10.7 +/- 2.0 vs. 8.3 +/- 2.3 cm/sec; 935.7 +/- 378.3 vs. 680.6 +/- 239.4 mL/min; 369.0 +/- 282.0 vs. 680.6 +/- 239.4 mL/min). Portal vein diameter, the congestion index (CI) of the portal vein, hepatic arterial resistance indexes, and the severity of esophageal varices did not differ between the two groups. In patients with a patent paraumbilical vein, the Child-Pugh score and the prevalence of ascites were significantly higher than in those without. In conclusion, the evaluation of PBV and PBF in cirrhotic patients can provide misleading results if a paraumbilical vein is patent, underestimating the degree of portal hypertension.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7489977 TI - The clinical significance of p53 gene mutation in hepatocellular carcinomas from Japan. AB - To clarify the clinical significance of the mutation of p53 gene in hepatocellular carcinoma (HCC), 90 resected specimens from Japanese patients were assayed using a polymerase chain reaction-single strand conformation polymorphism (PCR-SSCP) analysis. p53 mutations were detected in 25 cases (27.8%) at exons 4, 5, 6, 7, and 8, and the most frequent region of the mutation was at exons 5 and 7. No statistically significant correlation was observed between the p53 mutations and the clinical features except for the preoperative alpha-fetoprotein (AFP) level (P < .05). According to the pathological features, prognostic factors, such as size of the tumor, vascular invasion, fibrous capsule infiltration, and intrahepatic metastasis, showed no relationship to the existence of the mutation. However, p53 mutations were significantly associated with the degree of differentiation of HCC; that is, the mutation was found in 19 cases of 53 poorly differentiated HCCs (35.9%) and 2 of 3 cases of anaplastic HCCs (66.7%). The presence of p53 mutations was associated with a shortened cancer-free survival (P < .001, by log rank test) and a shortened survival (P < .05). A multivariate analysis by the Cox regression analysis showed that the p53 mutations were an unfavorable prognostic factor related to recurrence (P < .005), which is especially significant within the first postoperative year. These results suggest that the mutations of p53 gene of HCC might be an independent prognostic predictor to help in the selection of candidates who should undergo more intensive postoperative treatment. PMID- 7489976 TI - Renal effects of dietary supplementation with fish oil in cyclosporine-treated liver transplant recipients. AB - Nephrotoxicity is the main untoward effect of cyclosporine (CsA) treatment. Experimental and clinical data suggest that dietary supplementation with fish oil may lessen cyclosporine nephrotoxicity, possibly by lowering renal thromboxane (Tx) production. We have studied the renal effects of a daily supplementation for 2 months of 12 g fish oil (18% C20:5 n-3 eicosapentaenoic acid [EPA] and 12% C22:6 n-3 docosahexanoic acid [DHA]) in a placebo-controlled (12 g corn oil), prospective, randomized, double-blind study of stable CsA-treated liver transplant recipients. Thirteen patients ingested corn oil capsules and 13 fish oil. Compliance with dietary regimen was confirmed by fatty acid chromatography that showed increased plasma concentrations of EPA (from 0.4 +/- 0.02% to 4.6 +/- 0.5%, P < .0001) and DHA (from 1.8 +/- 0.2% to 3.9 +/- 0.1%, P < .0001) in the fish oil group and increased plasma concentration of linoleic acid (C18:2 n-6) in the corn oil group (from 25 +/- 2% to 28.4 +/- 2%, P < .001). At the end of the 2 months of the study, in the fish oil group the effective renal plasma flow increased by 22% (P = .012), the glomerular filtration rate increased by 33% (P = .057), the renal blood flow increased by 17% (P = .024), and the calculated total renal vascular resistances decreased by 20% (P = .034). In contrast, none of these parameters changed in the corn oil group. The renal functional reserve determined during L-arginine infusion, plasma renin activity (PRA), and plasma aldosterone (PA) remained unchanged during the study in either group.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7489978 TI - Expression of intercellular adhesion molecule 1 in human hepatocellular carcinoma. AB - We investigated the expression of intercellular adhesion molecule 1 (ICAM-1) in ex vivo human hepatocellular carcinoma (HCC) cells and in vitro in eight liver cancer cell lines, including six HCC cell lines and two combined hepatocholangiocarcinoma (CHC) cell lines. Immunohistochemistry showed the expression of ICAM-1 on the HCC cell surface with honeycomblike appearance in most cases (96.2%). On the other hand, hepatocytes in noncancerous areas did not express ICAM-1, except those hepatocytes in the periportal and intra-acinar areas with inflammation. Immunohistochemical study on cultured cells revealed that four cultured HCC cell lines and one CHC cell line constitutively expressed ICAM-1 on the cell surface and in the cytoplasm. Flow cytometric analysis revealed that immunostain-positive cells expressed surface ICAM-1 with more than a 90% positive cell rate, and their expressions were upregulated by incubation of cells with inflammatory cytokines, such as interferon alfa, interferon gamma, tumor necrosis factor-alpha, and interleukin 1 beta. Soluble ICAM-1 was detected in supernatants of cell lines expressing cell surface ICAM-1 expression, and was increased in amounts 2- to 20-fold by inflammatory cytokines. These findings suggest that liver cancer cells in ex vivo may express not only surface but also a soluble form of ICAM-1, differently from normal hepatocytes, and that both expressions are upregulated by inflammatory cytokines. PMID- 7489979 TI - The natural history of nonalcoholic fatty liver: a follow-up study. AB - Nonalcohol-induced fatty liver is widely believed to be a benign condition with little or no risk of disease progression. There have been occasional reports of progression to cirrhosis but none in the absence of preexisting fibrosis on the index biopsy specimen even when co-existing hepatitis was present (steatohepatitis). From our histological database (1978 to 1985), we identified 161 patients with fatty liver seen at our institution and traced the case notes of 156. One hundred five patients were initially excluded as having an alcohol induced cause, and the remaining 51 either were seen in the clinic (37) or had died, in which cases copies of their death certificates were obtained (14). A further 7 patients were excluded after clinic attendance gave evidence of alcohol excess and another 4 after review of their initial biopsy showed the presence of fibrosis or steatohepatitis. The apparent cause of the steatosis in the 40 included patients with strictly nonalcohol-induced pure fatty liver was obesity in 12, diabetes in 4 (1 obese patient), and cachexia associated with extrahepatic malignancy in 6. Four of the remaining 19 had serological evidence of an autoimmune disorder, but none of these had any clinical or histological features of autoimmune liver disease. Nine patients had evidence of hyperlipidemia, 3 of whom were also obese. At a median follow-up of 11 years (7 to 16), 12 of 26 living patients had abnormal results of liver blood tests and had repeat liver biopsies performed. None had progressed to steatohepatitis or cirrhosis; 1 obese patient had developed mild fibrosis 9.8 years after her index biopsy.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7489980 TI - Screening for hemochromatosis in children of homozygotes: prevalence and cost effectiveness. AB - Although hereditary hemochromatosis is an autosomal recessive disease, most homozygotes are concerned with the genetic implications for their children. The optimal age for testing children and the cost implications of screening their children have not been clearly established. A clinical database consisting of 255 children from families with at least one homozygote is used to assess the prevalence of homozygotes among children of homozygous parents and to review the biochemical abnormalities and life-threatening symptoms in these young adults. Decision analysis is used to estimate the cost and utility of screening children of a homozygous parent. Eleven homozygotes were discovered among children of homozygotes. Only one male had a life-threatening event, cirrhosis. Decision analysis estimated cost saving of $12 per child screened ($ net present value) and a saving of 10 quality-adjusted days per child screened at age 10 years compared with not screening. If screening began at age 20 years, there is a cost saving of $65 per child screened. Sensitivity analysis showed that the major factors influencing cost savings were the cost of venesections, sensitivity and specificity of the screening tests, and prevalence of disease. Because the prevalence of hemochromatosis is higher in children of homozygotes than in the general population, screening with transferrin saturation and ferritin as early as age 10 years is recommended. Savings are augmented if the cost per venesection is eliminated by allowing hemochromatosis patients to become voluntary blood donors. PMID- 7489982 TI - Deoxycholic acid influences cholesterol solubilization and microcrystal nucleation time in gallbladder bile. AB - Little is known about the effects of biliary deoxycholic acid on the partitioning of biliary cholesterol between vesicles and micelles and on the rate of nucleation of cholesterol microcrystals, key steps in gallstone formation. Therefore, 43 samples of fresh gallbladder bile were obtained from a heterogeneous group of patients with and without stones. Univariate and multivariate analyses were then applied to determine the inter-relationships between biliary cholesterol saturation, total lipid concentration, and bile acid species and (1) the distribution of biliary cholesterol between vesicles and micelles and (2) the cholesterol microcrystal nucleation time. The percentage of deoxycholic acid in bile was shown to be linearly related to the cholesterol saturation index (r = .54; P < .001), the vesicular cholesterol:phospholipid molar ratio (r = .53; P < .001), and the molar concentration of cholesterol in the vesicles (r = .59; P < .001). The mean proportion of biliary deoxycholic acid conjugates was also greater in patients with rapid nucleation times (23.4 +/- SEM 1.1%) than in those with slow nucleation times (17.3 +/- 1.9%; P < .05). As total bile lipid concentration increased, the proportion of total biliary cholesterol in vesicles decreased (r = .53; P < .001), whereas the molar concentration of vesicular cholesterol increased (r = .42, P < .01). The cholesterol saturation indices, total bile lipid concentration, and proportion of biliary deoxycholate were independent determinants of the molar concentration of cholesterol in vesicles. We conclude that relative increases in the percentage of deoxycholic acid and in bile lipid concentration, favor the partitioning of cholesterol into vesicles. In turn, this leads to an increase in the vesicular cholesterol:phospholipid molar ratio and thus to a decrease in the cholesterol microcrystal nucleation time. PMID- 7489981 TI - Evidence for the presence of an inactive precursor of human hepatocyte growth factor in plasma and sera of patients with liver diseases. AB - Human hepatocyte growth factor (hHGF), which is now known to be the same protein as the scatter factor and the tumor cytotoxic factor, is a heterodimeric protein with one heavy chain and one light chain linked together by a disulfide bond, and is thought to be involved in liver regeneration. Using an enzyme-linked immunosorbent assay (ELISA) and Western blot analysis, we found that a significant amount of single chain precursor of hHGF (pro-hHGF) was present in plasma of patients with fulminant hepatic failure (FHF) and that normal human serum contained a protease or proteases that convert pro-HGF to a heterodimeric (mature) form of hHGF. We also showed that the processing protease activity for hHGF was suppressed by such serine protease inhibitors as leupeptin, antipain, and aprotinin, and that sera of patients with liver diseases such as fulminant hepatic failure, acute hepatitis, chronic hepatitis, and cirrhosis contained not only pro-hHGF but also the protease. This is the first report showing the presence of pro-hHGF in human blood, and our observations suggest that hHGF is synthesized and secreted from the hHGF-producing cells as an inactive pro-hHGF after hepatic injuries, and the pro-hHGF is then converted to an active heterodimeric form of hHGF in the blood. It is also suggested that plasma of patients with liver diseases contains an active protease or proteases that convert pro-hHGF to a mature form of hHGF. PMID- 7489983 TI - Blunted natriuretic response to low-dose brain natriuretic peptide infusion in nonazotemic cirrhotic patients with ascites and avid sodium retention. AB - Patients with cirrhosis and ascites have high plasma levels of atrial (ANP) and brain (BNP) natriuretic peptides, two cardiac hormones released by the atria and ventricles, respectively. We evaluated renal hemodynamics, sodium excretion, and intrarenal sodium handling (lithium clearance method) in seven cirrhotic patients with ascites and avid sodium retention before, during, and after the infusion of synthetic human BNP, at the dose of 4 pmol/kg.min for 1 hour, which has been shown to increase renal plasma flow, glomerular filtration rate (GFR), and sodium excretion in healthy subjects without affecting systemic hemodynamics. Plasma BNP levels were 7.31 +/- 0.85 pmol/L in baseline conditions, and increased to 33.60 +/- 2.96 pmol/L at the end of the infusion (P < .01 vs. baseline). Urinary excretion of guanosine 3',5'-cyclic monophosphate (cGMP) also significantly increased during the infusion, indicating stimulation of natriuretic peptide receptors by BNP. BNP administration did not modify renal plasma flow, GFR, sodium excretion or tubular sodium reabsorption to any appreciable extent. Arterial pressure heart rate, plasma norepinephrine, and plasma renin activity (PRA) where also unchanged, whereas plasma aldosterone concentration showed a significant, 35% reduction at the end of the postinfusion period, ruling out the possibility that BNP-induced vasodilation might be responsible for failure of the peptide to induce a natriuretic response. Overactivity of antinatriuretic factors is probably the main determinant of the blunted natriuretic effect of BNP in these patients. PMID- 7489984 TI - The integrity of the cholecystokinin receptor gene in gallbladder disease and obesity. AB - Cholesterol gallstone disease and obesity are common and often associated disorders that could be affected by dysfunction of the receptor for cholecystokinin (CCK). Extending earlier studies that identified a defect at the level of receptor-G protein coupling in cholesterol gallstone disease, we characterized the primary structure of the gallbladder CCK receptor in patients undergoing a cholecystectomy. Represented were patients with cholesterol gallstones, as well as controls with pigment gallstones or without gallbladder disease. Both groups were composed of the range of body habitus from lean to morbidly obese. No evidence of any sequence mutation or polymorphism in the CCK receptor gene was found in any patient. This should lead future investigations of the pathogenesis of these problems toward the possible contribution of the plasmalemmal environment in affecting the association between normal receptors and G proteins. PMID- 7489986 TI - Disruption of circadian locomotor activity in rats after portacaval anastomosis is not gender dependent. AB - A recent study suggested that female rats are less affected by a portacaval anastomosis (PCA) than their male counterparts, as measured by body weight and changes in locomotor activity. In this study, we evaluated the entrainment of locomotor activity to the light/dark (LD) cycle, a consistent abnormality in the portacaval shunted rat. The degree of entrainment was measured in male and female rats before and after PCA or sham operation. All four groups of animals showed strong entrainment to an LD cycle before surgery. After portacaval anastomosis, male and female rats exhibited a highly significant decrease in overall motor activity as compared with the preoperative period and as compared with sham operated animals of the same gender. The percentage of total activity during daytime was significantly increased after portacaval anastomosis. The reduction in parameters of entrainment indicates a disruption of circadian function in both portacaval-shunted groups. Portal pressure measurements confirmed the patency of the shunts. Cortical brain glutamine levels were similarly increased in male and female shunted rats. The loss of body weight was slightly, but not significantly, more pronounced in male animals after shunt surgery. In conclusion, our results do not support a role for gender in the disruption of circadian function in rats after PCA. PMID- 7489987 TI - Regulation of gene expression by interleukin-6 in fetal rat hepatocyte primary cultures: role of epidermal growth factor and dexamethasone. AB - Fetal rat hepatocytes incubated in the absence of hormonal signals, or under proliferative (presence of epidermal growth factor [EGF]) or differentiative (presence of dexamethasone) culture conditions, showed responsiveness to interleukin-6 (IL-6). Northern blotting analysis for some typical acute phase genes such as haptoglobin and other proteins not previously identified as acute phase reactants, such as alpha-fetoprotein, beta 2-microglobulin, and fibronectin, showed a positive modulation by IL-6, in a dose-dependent manner. However, a well-characterized negative acute-phase reactant such as albumin was not responsive to IL-6. The well-established synergism between glucocorticoids and IL-6 on inducing transcription is absent in fetal hepatocytes. Conversely, the combination of IL-6 and EGF produced different patterns of expression, depending on the messenger RNA (mRNA) analyzed. Thus, EGF abolished the increased mRNA levels of haptoglobin caused by IL-6 but had no effect on other genes such as alpha-fetoprotein and fibronectin. PMID- 7489985 TI - Effect of donor age and sex on the outcome of liver transplantation. AB - We correlated donor and recipient factors with graft outcome in 436 adult patients who underwent 462 liver transplants. Donor variables analyzed were age, gender, ABO blood group, cause of death, length of stay in the intensive care unit, use of pressors or pitressin, need for cardiopulmonary resuscitation, terminal serum transaminases, and ischemia time. Recipient variables analyzed were age, gender, primary diagnosis, history of previous liver transplant, ABO blood group, cytotoxic antibody crossmatch, United Network for Organ Sharing (UNOS) status, and waiting time (except for the cross-match results, they were all known at the time of the operation). The endpoint of the analysis was graft failure, defined as patient death or retransplantation. Using multivariate analysis, graft failure was significantly associated with donor age, donor gender, previous liver transplantation, and UNOS 4 status of the recipient. The effect of donor age became evident only when they were older than 45 years. Livers from female donors yielded significantly poorer results, with 2-year graft survival of female to male 55% (95% CI, 45% to 67%); female to female, 64% (95% CI, 54% to 77%); male to male, 72% (95% CI, 66% to 78%); and male to female, 78% (95% CI, 70% to 88%). The only donors identified as questionable for liver procurement were old (> or = 60 years) women in whom the adverse age and gender factors were at least additive. However, rather than discard even these livers, in the face of an organ shortage crisis, their individualized use is suggested with case reporting in a special category. PMID- 7489988 TI - Acetaminophen-induced microvascular injury in the rat liver: protection with misoprostol. AB - Studies into the mechanism of acetaminophen (APAP)-induced hepatotoxicity have focused mainly at the hepatocellular level. This study aimed to investigate the effect of acetaminophen on the hepatic microvasculature using a vascular casting technique. Acetaminophen was administered at a dose of 650 mg/kg body weight (intraperitoneally) to fasted male Long Evans rats. Microvascular casting was performed at various points after drug administration. Liver casts from control rats showed good patency with normal hepatic microvasculature. Thirty-six hours after overdose with acetaminophen, liver casts showed rounded centrilobular cavities of various sizes, representing regions in which cast-filled sinusoids were absent with relatively normal microvasculature within periportal regions. Evidence of microvascular injury occurred as early as 5 hours after acetaminophen overdose. This injury consisted of changes to centrilobular sinusoids including areas of incomplete filling and dilated centrilobular sinusoids. Misoprostol (a prostaglandin E1 analog) treatment (6 x 25 micrograms/kg) given before and after acetaminophen administration markedly reduced the extent of microvascular injury with only small focal unfilled areas in the casts and a generally intact microvasculature. In conclusion, this study shows that overdosage with APAP resulted in an extensive, characteristic pattern of hepatic microvascular injury in the centrilobular region. The results also suggest that microvascular injury is an early event in the pathogenesis of acetaminophen hepatotoxicity. Misoprostol was found to protect against injury occurring at the microvascular level. PMID- 7489991 TI - Ciprofloxacin prevents the inhibitory effects of acute ethanol exposure on hepatic regeneration in the rat. AB - To determine whether the inhibitory effects of ethanol on hepatic regeneration could be prevented by ciprofloxacin, a fluroquinolone antibiotic with gamma aminobutyric acid (GABAA), receptor antagonist properties, adult, male Sprague Dawley rats (n = 6-8/group) received intraperitoneal injections of saline, putrescine (a hepatic growth promoter, 50 mg/kg), or ciprofloxacin (100 mg/kg), followed 1 hour later by gastric gavage with saline or ethanol (5 g/kg). One hour post-gavage, all rats underwent a 70% partial hepatectomy (PHx). Hepatic regenerative activity was documented 24 hours post-PHx by 3H-thymidine incorporation into hepatic DNA (DNA synthesis), proliferating cell nuclear antigen staining, and hepatic tissue putrescine levels. Compared with healthy controls, DNA synthesis rates were significantly lower in ethanol-gavaged/saline treated rats (22.7 +/- 4.4 x 10(3) vs. 12.3 +/- 6.9 x 10(3) DPM/mg DNA, respectively, P < .001) but unaltered in putrescine-(18.8 +/- 3.4 x 10(3) DPM/mg DNA) and ciprofloxacin-treated (18.3 +/- 2.6 x 10(3) DPM/mg DNA) rats. Hepatic proliferating cell nuclear antigen staining supported these findings. Hepatic putrescine levels also correlated with DNA synthesis data, being decreased in ethanol-gavaged/saline-treated rats (86 +/- 14 pmoles/mg tissue) compared with healthy controls (120 +/- 12 pmoles/mg, P < .01), ethanol-gavaged/putrescine treated (112 +/- 14 pmoles/mg, P < .05) and ethanol-gavaged/ciprofloxacin-treated (125 +/- 17 pmoles/mg, P < .05) rats. To determine whether these effects resulted from GABAA receptor-mediated changes in liver membrane potentials, intracellular membrane potentials were recorded before and 1 hour after PHx in healthy control, ethanol-gavaged/saline-treated and ethanol-gavaged/ciprofloxacin-treated rats.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7489989 TI - An array of mitochondrial alterations in the hepatocytes of Long-Evans Cinnamon rats. AB - In an attempt to identify the cellular targets of copper toxicity, we studied the ultrastructure of hepatocytes in the livers of 23 Long-Evans Cinnamon (LEC) rats ranging in age from 10 to 89 weeks. The hepatic copper concentration ranged from 325 to 2,126 (mean, 930) micrograms/g dry weight. Thirteen rats displayed varying degrees of jaundice at the time of killing. Numerous nuclei were indented by cytoplasmic invaginations. Conspicuous abnormalities were displayed by mitochondria. These included marked pleomorphism; increased or diminished matrical density; rearrangement, elongation, dilatation, stacking, or disappearance of cristae; absence of matrical granules; presence of matrical inclusions of flocculent electron-dense deposits; and degenerative changes. The severity of the ultrastructural changes did not correlate with the hepatic copper concentration but did correlate with the degree of the icterus displayed by the rats. Certain mitochondrial changes resembled those encountered in the hepatocytes of patients with Wilson's disease, confirming that these organelles are important targets of copper toxicity. PMID- 7489990 TI - Hepatoma cell-specific expression of a retrovirally transferred gene is achieved by alpha-fetoprotein but not insulinlike growth factor II regulatory sequences. AB - To target gene expression to malignant hepatic cells, we have constructed recombinant retroviral vectors containing a reporter gene encoding nuclear beta galactosidase (nls-LacZ) under transcriptional control of regulatory sequences from the rat alpha-fetoprotein (AFP) or human insulinlike growth factor II (IGFII) genes. The AFP and IGFII P3 promoters activate transcription during fetal development and are often reactivated in hepatocellular carcinoma (HCC). Infection of several cultured cell types with the retroviral vector containing the IGFII P3 sequence resulted in expression of the reporter gene in all cell lines tested, including those that do not produce IGFII. In contrast, selective expression was achieved by vectors containing the AFP transcriptional regulatory sequence. Nuclear beta-galactosidase activity was detectable in cells from lines that produce AFP, and not in cells that do not express the AFP gene. In most infected cell lines, retroviral RNA synthesis from the 5' LTR was inhibited, and deletion of the retroviral LTR enhancer did not change expression from either the IGFII P3-nls-LacZ or the AFP-nls-LacZ cassettes. After treatment of cells with 12 O-tetradecanoylphorbol-13-acetate and epidermal growth factor (EGF), the decrease in concentrations of endogenous AFP messenger RNA (mRNA) and nls-LacZ mRNA transcribed from the transferred AFP regulatory sequence were similar. In the context of an integrated provirus, the AFP transcriptional regulatory sequence is therefore subject to similar regulatory control as that of the endogenous gene. These data show that the AFP sequence, and not the IGFII P3 promoter we used, is suitable for targeting gene expression to malignant hepatic cells. PMID- 7489992 TI - Hepatobiliary transport of hepatic 3-hydroxy-3-methylglutaryl coenzyme A reductase inhibitors conjugated with bile acids. AB - To obtain prodrugs with affinity to liver parenchymal cells, the hepatic 3 hydroxy-3-methylglutaryl coenzyme A (HMG-CoA) reductase inhibitors HR 780 and lovastatin (syn. mevinolin) were conjugated with the bile acids cholic acid, taurocholic acid, and glycocholic acid. Hepatic uptake and biliary excretion of the coupled drugs were investigated and compared with the noncoupled drugs. Studies were performed with livers of normal Wistar rats, and TR-/GT- Wistar rats with deficient drug excretion. The experiments showed that the parent drug HR 780 was slowly excreted into bile. In contrast, the excretion of the bile acid conjugated HR 780 derivatives S 3554 (conjugated with cholate), S 3898 (conjugated with glycocholate), and S 4193 (conjugated with taurocholate) was rapid and very efficient in both groups of rat strains. The bile acid-conjugated HMG-CoA reductase inhibitors showed a 10 to 20 times higher affinity for the uptake systems of bile acids than the noncoupled parent drug compounds, and even higher affinities than the bile acids themselves. The cholate conjugate of HR 780 (compound S 3554) was shown to be a noncompetitive inhibitor of taurocholate uptake and a competitive inhibitor of sodium-independent cholate uptake (Ki = 1 mumol/L). Uptake of radiolabeled S 3554 into isolated rat hepatocytes was observed to be rapid, cell specific, saturable, energy dependent, and carrier mediated. However, the carrier for S 3554 uptake was found not to be the cloned Na(+)-dependent taurocholate cotransporting polypeptide Ntcp. Expression of this carrier cRNA in Xenopus laevis oocytes did not stimulate S 3554 uptake. PMID- 7489993 TI - Portal vein embolization with steel coils and absolute ethanol: a comparative experimental study with canine liver. AB - Portal vein embolization (PVE) is expected to have an adjuvant effect on the treatment of human liver tumors with transcatheter hepatic arterial embolization (TAE). In this experimental study, PVE was tried using two kinds of embolic materials, steel coils (coils) and absolute ethanol (ethanol). The aim of this article is to compare the effectiveness between the two methods. Five dogs and 12 dogs were treated with coils and ethanol, respectively. The second order portal branches were embolized. Embolization effects of each embolic material on the hepatic vessels and the liver parenchyma were studied angiographically and histologically. PVE with coils achieved immediate occlusion of the embolized vessels in all dogs. Unexpectedly, unembolized vessels were also constricted and/or obstructed in 4 of 5 dogs during a 4-week follow-up period. Histological examination showed no hepatic necrosis in all dogs. By contrast, in the dogs treated with ethanol, venous occlusion was achieved immediately after embolization with ethanol of 0.4 mL/kg or more. The embolized vessels remained occluded and no obstruction of unembolized vessels was observed until 4 weeks after embolization. Hepatic necrosis was observed in the segments where portal veins were occluded. The degree of parenchymal damage was proportional to the injected ethanol dose. Judging from these results, ethanol is likely to be more suitable than coils as an embolic material for PVE. PVE with coils is often dangerous because unembolized vessels might be occluded unexpectedly. For the clinical use of PVE with ethanol, the appropriate dose of injection should be investigated. PMID- 7489995 TI - Interleukin-1 and nitric oxide protect against tumor necrosis factor alpha induced liver injury through distinct pathways. AB - Mice sensitized with D-galactosamine (GalN) and challenged with recombinant murine tumor necrosis factor alpha (TNF alpha) developed severe apoptotic and secondary necrotic liver injury as assessed by histology, measurement of cytosolic DNA fragments, and determination of liver specific enzymes in plasma. Pretreatment with recombinant human interleukin-1 beta (IL-1) rendered mice insensitive to this TNF alpha toxicity. Coadministration of the liver-specific transcriptional inhibitor GalN with IL-1 prevented the development of tolerance, implicating de novo synthesis of liver specific proteins in the induction of tolerance. Pretreatment of mice with IL-1 resulted in elevated levels of nitrite/nitrate in serum and in enhanced nitric oxide synthase (NOS) activity in liver cells isolated from these animals. In addition, pharmacological doses of the nitric oxide (NO) donor sodium nitroprusside conferred complete protection against TNF alpha-induced liver injury in galactosamine-sensitized mice, suggesting a possible link between IL-1- and NO-induced protection. However, prevention of NO-synthesis by NG-monomethyl-L-arginine (NMMA) did not abolish IL 1-induced tolerance to TNF alpha in vivo. Cytotoxicity of TNF alpha to isolated hepatocytes sensitized with actinomycin D (ActD) was not significantly altered by inhibition of endogenous nitrite release. Also, enhanced NO production elicited in vitro by glycerol trinitrate or ex vivo by pretreatment with IL-1 had no significant effect in this system. We conclude that IL-1- and NO-induced protection of mice against TNF alpha-mediated liver damage follow distinct pathways. PMID- 7489996 TI - Bystander effect caused by suicide gene expression indicates the feasibility of gene therapy for hepatocellular carcinoma. AB - In the field of gene therapy using retroviral vectors, it appears impossible to introduce a foreign gene into all target cells. Therefore adjacent cell killing, the socalled bystander effect, caused by genetically modified cells provides therapeutic advantages for gene therapy against cancers. We retrovirally transduced the herpes simplex virus thymidine kinase (HSV-tk) gene into murine and rat hepatocellular carcinoma (HCC) cells. These HSV-tk gene-transduced HCC cells were cocultured with the corresponding parental cells in the presence of ganciclovir, at a concentration not at all cytotoxic to the parental cells. When parental HCC cells were cocultured with their HSV-tk gene-transduced counterparts at a high density at which most cells were in contact with one another, they were markedly eliminated. Conversely, when cocultured at a low density at which none of the cells were in contact, a weak but statistically significant bystander effect was observed. Addition of lysates of HSV-tk gene-transduced cells in the presence of ganciclovir did not cause and killing of parental cells. Furthermore, media conditioned by transduced cells with ganciclovir exhibited weak cytotoxic effects on parental cells. These results indicate that cell-cell contact plays a major causative role in the bystander effect and that minor contributors to this phenomenon are some cytotoxic substance released from transduced cells. Importantly, the bystander effect was induced in vivo as well as in vitro. When mixtures of transduced and untransduced HCC cells were implanted into the flank region of mice, intraperitoneal ganciclovir administration considerably inhibited tumor development, indicating the feasibility of gene therapy with HSV-tk gene and ganciclovir against HCC. PMID- 7489994 TI - Independent modulation of galactose-specific receptor expression in rat liver cells. AB - The expression of galactose-specific receptors on liver cells from rats at the end of pregnancy and from estrogen-treated animals was studied. The number and distribution of binding sites were estimated on hepatocytes and Kupffer and endothelial cells in vitro as well as in situ by means of protein-gold complexes. Hepatocytes and endothelial cells from pregnant rats showed an increased binding activity of at least three times for hepatocytes and one and a half times for endothelial cells with respect to normal rat livers. The increase in the hepatocyte receptor expression was paralleled by an increase in the level of its specific messenger RNA (mRNA). On Kupffer cells, a decreased number of binding sites, at least three times less than control values, was measured. The correlation between the altered hormonal level during pregnancy and the expression of galactose binding sites was examined in hepatocytes and Kupffer cells isolated from virgin rats treated with the synthetic estrogen diethylstilbesterol. In estrogen-treated rats both the binding sites and the specific mRNA of hepatocytes increased as compared with vehicle-treated or untreated animals. In contrast, in Kupffer cells both the estrogen treatment as well as vehicle-only injection led to a significant reduction in the expression of binding sites as compared with virgin untreated animals. To establish whether the decrease of galactose binding sites in Kupffer cells was related to the activation of macrophages or to the removal of plasma membrane caused by enhanced nonspecific phagocytosis, in situ binding experiments were performed after lipopolysaccharide (LPS)-stimulation or latex-bead phagocytosis. Nonspecific phagocytosis does not affect the binding activity, which instead appears strongly reduced after LPS injection. These findings suggest an independent response of galactose-specific receptor expression systems in the different types of liver cells to modulating agents. PMID- 7489997 TI - Gene expression of N-acetylglucosaminyltransferases III and V: a possible implication for liver regeneration. AB - N-acetylglucosaminyltransferases III and V (GnT-III and -V) are key enzymes in the synthesis of the branches of asparagine-linked oligosaccharides. Although their messenger RNAs (mRNAs) are expressed in various rat tissues, they are not detected in normal rat liver. Expression of the GnT-III and -V mRNAs, however, increased in regenerating liver after two-thirds partial hepatectomy compared with sham-operated rats. The enzymatic activities of GnT-III and -V increased in proportion to mRNA expression. To determine which type of cells in the liver have high activities of these glycosyltransferases, hepatocytes and nonparenchymal cells were separated by means of a two-step collagenase perfusion technique. GnT III activity was only detected in nonparenchymal cells of normal rat liver. However, during liver regeneration, GnT-III activity increased and thus was also detectable in hepatocytes. GnT-V activity was detected in both types of cells. These data were supported by reverse transcription-polymerase chain reaction results. Although the mechanism underlying the induction of these glycosyltransferases is unknown, lectin blot analysis showed that oligosaccharides in many glycoproteins, including hepatocyte growth factor, a major growth factor associated with liver regeneration, were newly synthesized during liver regeneration. This is the first report on the expression of glycosyltransferases during liver regeneration and suggests that there are different mechanisms involved in regulation of the genes of GnT-III and -V during liver regeneration. PMID- 7489998 TI - Endothelium-dependent vascular hyporesponsiveness without detection of nitric oxide synthase induction in aortas of cirrhotic rats. AB - The present experiments were designed to test if induction of nitric oxide synthase (NOS) plays a role in the systemic vasodilation observed in hepatic cirrhosis. Because endotoxin levels are elevated in cirrhosis, and endotoxin stimulates inducible nitric oxide synthase (iNOS) expression in several cell lines, aortas of carbon tetrachloride-induced cirrhotic rats with ascites were evaluated for iNOS expression. Endotoxin-treated rats were studied as positive controls. Phenylephrine contraction was decreased in aortic rings with endothelium from both endotoxin-treated and cirrhotic rats as compared with controls. However, after endothelium denudation, the reduced contractility persisted in endotoxin-treated rats but disappeared in cirrhotic rats. L-Nitro arginine-methylester (L-NAME), a nonselective inhibitor of NOS, potentiated the phenylephrine contraction of aortic rings with and without endothelium from endotoxin-treated rats but only rings with endothelium from cirrhotic rats. Moreover, aminoguanidine (AG), a preferential inhibitor of iNOS, did not affect phenylephrine contraction of rings with or without endothelium from cirrhotic rats but reversed the blunted response in endotoxin-treated rats. Northern analysis detected iNOS RNA (mRNA) expression in aortas of endotoxin-treated rats but did not detect it from cirrhotic rats. In summary, although several previous studies provide evidence for in vivo overproduction of nitric oxide in cirrhosis, the present results do not support iNOS induction as the source of nitric oxide in aortas of cirrhotic rats. Rather, because the aortic vascular hyporesponsiveness in cirrhosis is endothelium-dependent, overexpression or overstimulation of the endothelial constitutive isoform of NOS appears to be involved. PMID- 7489999 TI - Treatment of chronic type B and C hepatitis with interferon alfa: an economic appraisal. AB - The aim of this study is to assess the long-term economic impact of treatment of chronic hepatitis B and C with interferon alfa. Estimates were made of the progression of the disease over a 30-year period using a transitional probability model. Cohorts of 1,000 hypothetical patients with either chronic hepatitis B or C treated with interferon alfa were compared with an untreated cohort. The costs were estimated for therapy, monitoring, and treatment of the disease, including transplantation. The cost-effectiveness of therapy was expressed in terms of cost per life saved, cost per year of life saved, and cost per quality-adjusted year of life saved. The analysis was extended to include the indirect costs to patients. The analysis included two rates of progression, two mortality rates, and discounted and undiscounted costs. Mortality in the treated group was lower, saving 18 to 31 lives in the hepatitis B virus (HBV) cohort and 13 to 22 lives in the hepatitis C virus (HCV) cohort. Fewer patients progressed to cirrhosis or decompensated cirrhosis. Discounted costs per year of life saved ranged from 2,142 pounds to 17,128 pounds. A cost-benefit analysis indicated excess benefits over costs when values for life were included in the analysis. The potential usefulness of interferon alfa on the clinical and economic outcome of treatment is indicated from the model. These findings together with the benefits that are likely to accrue from the reduction in infectious individuals suggest that this therapy has a role to play in public health policy to contain the impact of hepatitis. PMID- 7490000 TI - Chronic hepatitis and hepatic failure in a 14-year-old girl. PMID- 7490001 TI - Economic modeling of treatment in chronic hepatitis B and chronic hepatitis C: promises and limitations. PMID- 7490002 TI - Hepatocellular carcinomas associated with hepatitis B and C virus infections: are they any different? PMID- 7490003 TI - Keratinocyte growth factor--just another mitogen or the "Holy Grail" that regulates liver regeneration? PMID- 7490004 TI - Can a guest liver bring the gift of a gene to disarm a hostile host? PMID- 7490005 TI - Infusions of diazepam and allopregnanolone into the midbrain central gray facilitate open-field behavior and sexual receptivity in female rats. AB - We have previously shown that blocking GABAA receptors with bicuculline in the dorsal and lateral quadrants of the rostral two-thirds of the midbrain central gray (MCG) in rats reduces the lordosis quotient (LQ) and proceptive behaviors without increasing the defensive index (McCarthy, Pfaff, and Schwartz-Giblin, 1991b), indicating that tonic release of endogenous GABA in the MCG has a primary facilitatory effect on reproductive behaviors. The present study tests whether two modulators of the GABAA-benzodiazepine-receptor complex, diazepam and the progesterone metabolite, allopregnanolone, facilitate female receptivity as well as open-field behavior. Ovariectomized estradiol benzoate (EB)-treated rats were bilaterally infused in the MCG with vehicle, diazepam, or allopregnanolone and tested 20 min after diazepam and 12 min after allopregnanolone. Animals were tested once each for both receptive or open-field behavior with testing sessions being at least 1 week apart in a crossover design between vehicle and one of the two drugs. Both diazepam and allopregnanolone significantly increased the lordosis quotient in a dose-dependent manner, the lowest effective dose tested being 100 and 250 ng, respectively. In the open-field test, infusion of diazepam (100 ng) in EB-treated rats significantly increased the ratio of central squares entered to total squares entered (C/T) without a change in total activity. Rats without EB elicited comparable C/T ratios when tested with five times as much diazepam. Diazepam infusions at more dorsal sites in the midbrain and in overlying cortex were without effect on C/T ratios in EB-treated rats. Allopregnanolone (500 ng) infusions in MCG of EB-treated rats did not alter C/T ratios compared to animals without EB. Our results are consistent with diazepam and allopregnanolone acting through the GABAA receptors of the MCG to enhance female sexual receptivity and with estradiol potentiating the effect of diazepam in the MCG to increase open-field behavior. PMID- 7490006 TI - The effects of neonatal treatment with tamoxifen on sexually dimorphic behavior and morphology in gray short-tailed opossums (Monodelphis domestica). AB - The effects of postnatal exposure to the antiestrogen tamoxifen on sexually dimorphic behavior and morphology in adult male and female gray short-tailed opossums were examined. Following gonadecomy and treatment with estradiol in adulthood, neonatally tamoxifen-treated (TX) females showed less female-typical screeching threat behavior and TX males showed less male-typical precopulatory behavior than same-sex controls. While testes failed to descend and phalluses were shorter in juvenile TX males than in juvenile control males, behavior did not differ between TX and control animals following treatment with testosterone in adulthood. The findings that activity level following treatment with estradiol in adulthood and body weight in juveniles and adults were significantly higher in TX than in control animals of both sexes suggest that perinatal estrogens are involved also in the organization of these characteristics. The significance of these findings is discussed with respect to the development of sex differences in behavior and morphology in eutherian mammals. PMID- 7490007 TI - Estrogen replacement therapy may protect against intellectual decline in postmenopausal women. AB - Postmenopausal women over the age of 50, who were either on estrogen replacement therapy (N = 21) or not (N = 33), were assessed on several sexually dimorphic cognitive functions. The two groups were strictly equivalent in age, education, and vocabulary score (an abbreviated measure of past intelligence). Overall, women on therapy had better scores than those not on therapy, regardless of whether the specific function favored men or women. Previous history of reproductive surgery had no effect on scores. Estrogen may guard against some of the intellectual decline which is to be expected postmenopausally. PMID- 7490009 TI - The medial amygdaloid nucleus and medial preoptic area mediate steroidal control of sexual behavior in the male Syrian hamster. AB - An important goal of studies on steroid receptors in the brain is to understand the functions of specific populations of steroid receptor-containing neurons, particularly in the control of sexual behavior. The present study compared the ability of testosterone implants directed toward the medial amygdaloid nucleus (Me) or the bed nucleus of the stria terminalis (BNST) and medial preoptic area (MPOA) to stimulate mating in castrated males. Twenty adult male hamsters were selected for vigorous sexual behavior, and baseline measures of copulatory behavior during the first 10 min of a 30-min mating behavior test were recorded on two occasions. Twelve weeks after castration, sexual behavior was recorded as before, and each male received a single intracranial implant constructed of 23 gauge tubing packed with crystalline testosterone placed stereotaxically into Me or BNST/MPOA (n = 10 each). Behavior was assessed on two occasions after surgery to determine if implants in Me or BNST/MPOA could stimulate sexual behavior above that observed after castration. In half of the males from each group, testosterone increased the males' interaction with the female, anogenital investigation, mounting, and reduced the latency to the first mount. After completion of behavioral testing, males were perfused and the brains processed for androgen receptor immunocytochemistry to determine the extent of brain regions influenced by the testosterone-filled cannula. In tissues stained rapidly in diaminobenzidine, androgen receptor-containing neurons were visible only in steroid responsive brain regions adjacent to the testosterone implant. This approach revealed that steroid receptors in both the posterior subdivision of Me and in the medial subregions of BNST/MPOA can mediate hormonal control of mating behavior in the male Syrian hamster. Implants placed outside these regions failed to stimulate mating. These results suggest that maintenance of copulatory behavior by gonadal steroids is not transduced by a single brain region, but that steroid effects can be elicited at multiple points along the mating behavior pathway. PMID- 7490008 TI - Effects of neonatal testicular suppression with a GnRH antagonist on social behavior in group-living juvenile rhesus monkeys. AB - Twenty-four male and eight female 1-year-old rhesus monkeys (Macaca mulatta) were observed for social interaction with other yearlings and with their mothers. The males comprised three groups which differed in the level of neonatal androgen exposure. One group received the GnRH antagonist Antide during their first four neonatal months (Antide n = 8), which suppressed pituitary LH secretion resulting in peak neonatal T levels < 0.7 nmol/liter. A second group received Antide treatment combined with a long-lasting testosterone replacement (Ant/And n = 8), which resulted in peak neonatal T levels of 29.1 +/- 3.8 nmol/liter. The third group (Vehicle n = 8) received the Antide and androgen vehicles and had intermediate peak T levels of 5.2 +/- 1.0 nmol/liter. Behavior of males was compared to that of unmanipulated control females living in the same social group (Control Female n = 8) when androgen levels were uniformly low (< 0.7 nmol/liter) in all male groups. Subjects received 12 weekly 30-min focal observations by an observer blind to the neonatal treatments. Marked sex differences were found in several categories of sociosexual behavior. All three groups of males engaged in significantly more sexual and play behavior than females, with the exception of quiet solitary play, which females exhibited significantly more frequently In addition, females exhibited significantly more interest in infants than did any male group. There were no differences between groups in agonistic behavior or time spent in contact with other individuals, but females spent significantly more time than any male group in proximity to other animals. Both females and Antide males initiated proximity and followed animals significantly more frequently than Ant/And males, but not Vehicle males. Proximity durations with mothers initiated and terminated by yearlings were longer for females than for any male group and for Antide males than for Ant/and males. Antide males were groomed significantly longer than any other group. These results demonstrate effects of neonatal testosterone exposure on social behavior in yearling rhesus. Suppression of neonatal T did not affect sexually dimorphic patterns of play and sexual behavior, but altered the character of interactions with their mothers. Whether this reflects a delay in the development of maternal independence or a fundamental alteration in patterns of social interaction remains to be resolved. PMID- 7490010 TI - Hormones, sex, and status in women. AB - Androgens are often associated with assertive behavior; under what circumstances is this reflected in higher dominance rank? In this study of coresidential college women, androgens (total testosterone, free testosterone, and androstenedione) and estradiol were positively correlated with high self-regard in women (as measured by the degree to which subjects over-ranked themselves in a peer-ranking task) and with infrequent smiling, a behavior that has been associated with dominance in previous studies. Androgens and estradiol were also positively correlated with number of sexual partners. The behaviors engendered by these hormones are often positively correlated with high dominance rank, at least in males. In this population, however, high rank (as judged by peer assessments) was negatively correlated with androgens, particularly androstenedione, and showed a negative trend with estradiol as well. One possible interpretation of these findings is suggested by an evolutionary perspective that sees different routes to status among women who compete for resources directly and women who obtain resources through investing males. PMID- 7490011 TI - Endocrine basis for two types of individual differences in lordosis reflex intensity. AB - We have identified individual differences in two independent types of lordosis reflex intensity. Overall lordosis reflex intensity, averaged across 9.4 +/- 0.99 days (range = 1-18 days), differs among cycling individuals and is a biomarker for the rate and pattern of reproductive aging. Although estrogen is necessary to permit lordosis reflex responding and manifests individual differences throughout the estrous cycle, it was individual differences in progesterone that correlated negatively with the individual variation in overall lordosis reflex intensity; females with intense overall lordosis reflexes had low progesterone throughout the estrous cycle. Independent of their overall reflex intensity, females also displayed individual variation in lordosis reflex intensity on proestrus. Among females that still maintained a cyclic lordosis reflex, those with intense lordosis reflexes on proestrus had high levels of luteinizing hormone (LH) at lights out. Among females that displayed intense lordosis reflexes throughout the estrous cycle (i.e., had entered constant lordosis), those with intense proestrous lordosis reflexes had low levels of LH at lights out on proestrus. Females who had entered constant lordosis tended to have lower overall progesterone than females who still maintained cyclic patterns of lordosis. Thus, constant lordosis was identified as an important component of the changes of reproductive aging. PMID- 7490013 TI - The relationship of cortisol levels to social environment and reproductive functioning in female cotton-top tamarins, Saguinus oedipus. AB - Cortisol levels were examined in 17 cotton-top tamarin monkeys (Saguinus oedipus) to determine: (1) if first-morning void urine sampling could be used as a noninvasive method for monitoring cortisol excretion, (2) if capture and bleeding were associated with changes in urinary cortisol levels, (3) the relative cortisol levels in reproductively active and reproductively suppressed females, and (4) the relationship between cortisol levels and changes in social condition in cotton-top tamarins. Mean urinary cortisol levels during ovarian cycling did not differ between captured and bled females and undisturbed females. Mean cortisol levels were significantly lower in females who were housed in their natal groups and reproductively suppressed than in the same females when they were removed and placed adjacent to a novel male and ovarian cycling began. For all females, mean cortisol levels were higher during the periovulatory period than during the nonperiovulatory period of the ovulatory cycle, with mean cortisol levels higher in newly cycling females than in long-term cycling females. No differences were found in mean cortisol levels between long-term cycling females and prepubertal females and postpubertal natal females. Cortisol levels from long-term cycling females were much lower than during the first two to three ovarian cycles occurring in newly cycling female tamarins, during the last part of pregnancy, and during the first 6 weeks postpartum. These data provide evidence that (1) suppression of ovarian cycling in postpubertal females remaining in their natal family is not associated with stress-induced changes in cortisol levels, (2) cortisol levels are not strictly tied to reproductive condition, and (3) social change may be reflected in cortisol elevation. PMID- 7490012 TI - Investigation into the role of cholecystokinin (CCK) in the induction and maintenance of maternal behavior in rats. AB - The biochemical regulation of maternal behavior has been extensively studied. Cholecystokinin (CCK), a gut peptide that is also present in the brain, recently has been implicated in the onset of maternal behavior in estrogen-primed virgin rats. The objective of the present set of studies was to delineate the role of CCK in the onset (Experiments 1-3) and maintenance (Experiments 4 and 5) of maternal behavior in rats. In the first study intracerebroventricular (i.c.v.) administration of CCK was unable to stimulate the onset of maternal behavior in estrogen-primed virgin rats. Similarly, i.c.v. infusions of CCK into pregnant rats, starting on Day 17 of gestation (Experiment 2), did not advance the onset of maternal behavior. Moreover, when CCK-filled minipumps were implanted intraperitoneally in estrogen-primed virgin rats, the rate of onset of maternal behavior was unaffected (Experiment 3). In contrast, direct infusions of CCK into the MPOA blocked the disruptive effects of beta-endorphin on the maintenance of maternal behavior in postpartum lactating rats (Experiment 4). In addition, proglumide, a CCK receptor antagonist, disrupted maternal behavior in postpartum lactating rats by increasing latencies to retrieve and crouch over the young (Experiment 5). These results support an involvement of CCK in the maintenance, but not the onset, of maternal behavior in rats. PMID- 7490014 TI - [Simplified enzyme-linked immunoadsorbent assays for myelin basic protein and antibodies to myelin basic protein]. AB - Synchronous quantitative determination using simplified enzyme-linked immuno adsorbent assay has been developed for myelin basic protein (MBP) and antibodies to myelin basic protein (Anti-MBP). Antisera were prepared by immunizing rabbits with purified MBP from human white matter. Antibodies to MBP was conjugated with horseradish peroxidase (HRP) by using modified periodate oxidation method. Minimum detection amounts of MBP and Anti-MBP are 0.25 ng/ml and 0.20 ng/ml respectively. The activities of Anti-MBP-HRP conjugates are stable (stored at -30 degrees C) for at least 2 years. The detection procedure can be accomplished within 24 hours. PMID- 7490016 TI - [The value of serum myelin basic protein in assessment of severity of acute closed head trauma]. AB - Serum levels of myelin basic protein (MBP) were measured in 112 patients after acute head injury by enzyme linked immuno-absorbent assay (ELISA). Patients with cerebral concussion showed no significant change in serum MBP. Patients with cerebral contusion, extradural hematoma or intracerebral hematoma had a mean serum MBP concentration much higher than that of patients with cerebral concussion. And patients with extradural hematoma had a mean serum MBP concentration much lower than that of patients with cerebral contusion and intracerebral hematoma; the differences were significant (P < 0.05). But there was no significant difference between patients with cerebral contusion and intracerebral hematoma (P > 0.05). The amount of serum MBP was significantly correlated with the volume of extradural hematoma, intracerebral hematoma and with the extent of the cerebral contusion (P < 0.05). This study suggested that the serum MBP may reflect the type and severity of closed head injury patients. PMID- 7490015 TI - [Enzyme-linked immunoadsorbent assays for myelin basic protein and antibodies to myelin basic protein in serum and CSF of patients with diseases of the nervous system]. AB - Simplified enzyme-linked immunoadsorbent assays (ELISA) for myelin basic protein (MBP) and antibodies to myelin basic protein (Anti-MBP) have been used to test 337 patients with diseases of the nervous system including 36 compressive diseases (CMP), 33 multiple sclerosis (MS), 34 cerebrovascular diseases (CVD), 31 inflammatory diseases of central nervous system (ID), 161 epilepsy (EP) and 42 other nervous diseases (OND). Comparison of results among various groups indicates that serum mean MBP values of CMP, MS, CVD, ID and EP groups are significantly higher than those of OND group and normal control (P < 0.01). The serum mean MBP value of 33 acute trauma patients with spine fracture and paraplegia, the majority of CMP group, is the highest compared with MS group (P < 0.05), CVD, ID and EP groups (P < 0.01). CSF mean MBP value of 15 CVD patients is markedly greater than that of OND group (P < 0.05). No statistically significant differences are found in serum MBP values between OND group and normal control, and in serum and CSF Anti-MBP values among six groups by using our method. PMID- 7490017 TI - [Myelin basic protein level in serum of patients with chronic cor pulmonale in acute phase]. AB - With a sensitive enzyme-linked immunoadsorbent assay that can detect as little as 0.5ng/ml of myelin basic protein (MBP), we studied serum specimens from 34 patients with chronic cor pulmonale (CP) and 33 patients with chronic bronchitis (CB) during acute attack in hospital and 30 control subjects. The results showed that the serum mean MBP level of CP patients was markedly higher than those of CB patients and controls (P < 0.01), but no statistically significant differences were found between CB patients and controls (P > 0.05). There was a positive correlation between elevated levels of serum MBP and carbon dioxide partial pressure (PaCO2). This suggested that elevated serum MBP levels may be associated with brain destruction by hypercapnia and hypoxia of CP patients. PMID- 7490018 TI - [Correlation analysis of lipids and apolipoproteins in plasma HDL, LDL and VLDL of normal subjects and endogenous hypertriglyceridemics]. AB - The linear correlation and multiple stepwise regression analysis of lipids and apolipoproteins in plasma, HDL, LDL and VLDL of 20 normal subjects and 20 patients with endogenous hypertriglyceridemia was made. The result of simple factor analysis showed that there was a positive correlation of TG and VLDL TG with apoC II, C III, E, LDL apoB100 and VLDL apoC II, C III, E, B100; and a negative correlation with HDL apoA I. HDL-C was related positively with apoA I, HDL apoA I, C III and negatively with apoC III, E, LDL apoB100 and VLDL apoB100, C II, C III, E. TC was correlated positively with apoB100 and LDL apoB100. The result of multiple factor analysis showed that apoC III, E, B100/A I, VLDL apoC III and HDL apo C II, E, C II/C III, among 20 factors, were main factors which were related with TG. Apo C III, VLDL apoC III and LDL apoB100 were main factors which affected VLDL TG. ApoB100, and C II were main apolipoproteins which were related with TC; while the main apolipoproteins which affected HDL-C were apoA I, HDL apoA I, C II, VLDL apoE, HDL apoA II, the significance of the results is discussed. PMID- 7490019 TI - [Effect of native and oxidation-modified LDL, VLDL and HDL on the morphological appearance and phenotype of cultured human arterial smooth muscle cells]. AB - The arterial smooth muscle cells (SMC) are the predominant type of cells in atherosclerotic lesions, and their proliferation plays an important role in the process of AS genesis. We established the primary culture and sub-culture method for human arterial SMC and observed the effects of LDL, VLDL, HDL, OX-LDL, OX VLDL and OX-HDL on the morphological appearance and phenotype of cultured human arterial SMC. The results revealed that the cells cultured in the medium containing OX-LDL, OX-VLDL and OX-HDL (25 micrograms/ml) became more irregular with long finger-like projects and lacked a bipolar shape, and the characteristic "hills" and "valleys" disappeared. LDL and VLDL also had the influence but in smaller degrees. HDL did not have any effect. The SMC cultured in the presence of OX-LDL (25 micrograms/ml) had less myofilaments and more endoplasmic reticulua, Golgi and mitochondria in cytoplasm compared with the cells in control group under electronmicroscope. These results suggest that the atherogenic role of LDL, VLDL, OX-LDL, OX-HDL, and OX-VLDL are closely related to their stimulating effects on the modulation of phenotype of arterial SMCs from "contractile" towards "synthetic". PMID- 7490020 TI - [The effective time of HX solution for rat liver preservation]. AB - The effects of HX solution and UW solution for rat liver preservation were studied with non-circulated isolated perfused rat liver model. Sixty-six inbred Wistar rat livers were randomly preserved for 12, 18, 24, and 30h with HX solution or UW solution. The effects were assessed by measuring hepatic tissue water content, sinusoidal lining cell mortality, Krebs-Henseleit's perfusate aspartate aminotransferase, the number of livers secreting bile during isolated perfusion, and by observing the morphological changes in liver. The experimental results suggest that the effects of HX solution for rat liver preservation be similar to those of UW solution within 24h storage. PMID- 7490021 TI - [Effect of 153Sm-EDTMP on hematopoiesis and vital organs of 93 patients with bone tumor]. AB - In the present study data on blood cell count, serum biochemistry, electrolyte, enzyme and vital organs of 93 patients with bone tumor or metastasis were investigated before and after treatment with 153Sm-EDTMP. The results showed that, 7 days and 30 days after the administration of 153Sm-EDTMP (< 29.6 MBq) (0.8 mCi/kg), the levels of hemoglobin, WBC lymphocyte count, and the liver and kidney function of all patients were not significantly different from the baseline data before treatment (P > 0.05). Although at 7 days, there was a declination of the granulocyte count, it returned to normal at 30 days (P > 0.05). The platelet count was significantly decreased (0.05 > P > 0.01). at 30 days after the administration of 153Sm-EDTMP. Thirteen patients received 74-185 MBq (2-5mCi/kg) and their myelo biopsies at 3 and 18 months showed no sign of acute or chronic toxicosis. PMID- 7490022 TI - [Determination of modulus of elasticity of human periodontal membrane]. AB - The modulus of elasticity of periodontal membrane, as an essential parameter indicating the mechanical character of periodontal membrane, is very important in biomechanical studies (e.g. finite element stress analysis and photoelastic analysis) of dental restoration. However, determination of modulus of elasticity of human periodontal membrane has not been reported in China, and there are differences among the foreign reports. In this study, 135 periodontal samples were collected from fresh teeth and alveoli of 6 healthy adults. Drawing and compressing test were performed. The results showed that the tensile modulus of elasticity of human periodontal membrane is between 3.35 and 4.59MPa and the compress modulus of elasticity 0.5402(+/- 0.348) MPa. PMID- 7490023 TI - [Adsorption of aclacinomycin A onto polyisobutylcyanoacrylate nanoparticles]. AB - The mechanism of adsorption of aclacinomycin A onto polyisobutylcyanoacrylate nanoparticle and the factors effecting the adsorption were studied. The adsorption process reached equilibrium in 15 minutes. The results showed that the surface charge densities of the nanoparticles, pH, ionic strength and temperature effected the adsorption. Adsorption isotherms of aclacinomycin A onto polyisobutylcyanoacrylate nanoparticles could be described with Freundlich equation Y = 1.0188 C0.6494 (r = 0.9945) and with Langmuir equation C/Y = 0.7363C + 0.4027 (r = 0.9331) respectively. The main actions between aclacinomycine A and polyisobutylcyanoacrylate nanoparticles were electrostatic and ionic action. PMID- 7490024 TI - [Study on the lung targeting gelatin microspheres of streptomycin sulphate]. AB - This paper is reported the technology of lung targeting gelatin microspheres of streptomycin sulphate (SMS). The microspheres were prepared with natural biodegradable gelatin as the load material and castor oil as the oil phase. The experimental conditions were optimized, the mean volume diameter obtained being 9.7 microns and the mean rate of encapsulation 15.69%. The content, shape and size of the microspheres showed no remarkable change after storage at 37 degrees C RH 75% for 3 months. Activation energy of heat decomposition E = 75.86kJ/mol. In vitro, the SMS release rate was found to accord with Higuchi equation with t1/2 = 8.6h. In vivo (rabbits) the gelatin microspheres were proved to be concentrated in the lung. PMID- 7490025 TI - [Physicochemical characteristics of gentamicin polybutylcyanoacrylate nanoparticles]. AB - Several formulations of gentamicin nanoparticle (GM-NP) colloids were prepared by the emulsion polymerization technique using the polybutylcyanoacrylate as the carrier. Various kinds of physicochemical characteristics, such as particle size and size distribution, drug loading and associating ratio, surface zeta potential, surface tension, turbidity, relative density, viscosity, refractive index and acidity were observed, determinated and compared. The results of the experiments have contributed to a full understanding of the physicochemical characteristics of gentamicin nanoparticles and also provided a basis for the establishment of the quality evaluation methods. PMID- 7490026 TI - [Effect of tetramethylpyrazine on lymphocytes proliferation response of murine splenocytes]. AB - The effect of tetramethylpyrazine (TMP) on lymphocytes proliferation response (LPR) detected by 3H-TdR incorporation into murine monocytes. The results showed that TTMP at the concentration of 100-1000 micrograms/ml inhibited LPR in normal mice and B14-melanoma bearing mice in vitro the effect was dose-dependent. It was also found TTMP could stimulate LPR to Con A at the concentration of 1 micron/ml in normal mice, and at the concentration of 100-1000 micrograms/ml in B16 melanoma bearing mice. But TTMP inhibited LPR to Con A at the concentration of 100-1000 micrograms/ml in normal mice. The restoration of LPR to PHA and increasing 3H-TdR incorporation into splenocytes were found in vivo in B16 melanoma bearing mice after treatment with TTMP. The data indicates that TTMP has an immune modulation. PMID- 7490027 TI - [Rheologic properties of the human umbilical vein endothelial cells exposed to shear stress]. AB - The function of vascular endothelial cells has recently become a hot spot in biomedical research. This paper reports the deformation properties of cultured human umbilical vein endothelial cells exposed to shear stress with the flow chamber. The umbilical vein endothelial cells were exposed to shear stress 0.6, 3.8 and 7.5 Pa for 12 h respectively. With microvideo recording and computerized morphologic analysis, the results demonstrated that the endothelial cells were elongated as a result of exposing to shear stress 3.8 or 7.5 Pa for 12 h. Their major axes were consistent with the flow direction. Furthermore the degree of elongation and orientation of endothelial cells depended on the shear stress and shear duration. The results were compared with those of three foreign laboratories; and the causes of agreement and difference were discussed. This study is the basic word for a series of future researches. PMID- 7490028 TI - [A new and simple method of evaluating left ventricular contractility and its application]. AB - After making theoretical analysis, we detected the relationship among the maximal tension of ventricular contraction, left ventricular end-systolic pressure and stroke volume, and then derived a new and simple method of evaluating left ventricular contractility, the slope of the left ventricular end-systolic pressure-volume relation (Emax). Twelve mongrel male dogs were used to test this method. The results show that the value of Emax (0.60 +/- 0.16 kPa/ml) coming from this simple method is very close to that from complicated method; this method has good repetitiveness and is very sensitive to the variation of myocardial contraction. Using the method for evaluating left ventricular contraction after ischemia, we found the Emax more cogent than other hemodynamic parameters. Therefore, this method is an efficient method of evaluating left ventricular contraction. PMID- 7490029 TI - [Analysis of pressure-volume-hysteresis in rabbit]. AB - The pressure-volume (PV) relationships of respiratory, lung and chest wall (Prs V, PL-V and Pcw-V) were studied in 11 white rabbits. The relaxation pressure induced by Hering-Breuer reflex was measured on inflating certain volume of air. Four groups of inflation experiments were performed in each rabbit. (1) Inflations of incremental tidal air with 10 ml in each increment were performed from functional residual capacity (FRC) to 70 ml, then inflations of decremental tidal air with 10 ml in each decrement were done from 70 ml to FRC. (2) 70 ml of air was inflated just before each incremental and decremental inflation. (3) 10 ml more air was inflated before each predicted incremental inflation. (4) The first group inflation was repeated. THE RESULTS: The P-V curves showed a "S" shape in all 4 groups. There was an obvious hysteresis on the Prs-V and PL-V curves in the 1st group between the incremental and decremental inflations; it disappeared in the 2nd and 3rd groups. Hysteresis reappeared in the 4th group, but it was smaller than that in the 1st group. These results substantiate the theory proposed by Frazer that pulmonary hysteresis mainly comes from sequential opening and closing of lung units during inflation and deflation. PMID- 7490030 TI - [A study of c-src gene express product pp60c-src in esophageal carcinoma]. AB - Using the specific monoclonal antibody of c-src gene express product, pp60c-src (McAb 327), we studied immunohistochemically in this experiment 35 cases of the esophageal squamous cell carcinomas and 22 adjacent carcinoma tissues. The positive rates of pp60c-src in the squamous cell carcinomas of the esophaguses, the proliferative and the normal squamous epithelis adjacent to the carcinoma were 65.7% (23/35), 100.0% (15/15), and 57.1% (4/7), respectively. There was significant difference among them statistically (P < 0.05). And the express amount of pp60c-src in the esophageal squamous cell carcinoma was higher than those in the proliferative and normal squamous epithelia adjacent to the carcinoma. The positive rates of pp60c-src of the esophageal squamous cell carcinoma with different differentiation grades I, II and III were 80.0% (8/10), 68.4% (13/19), and 33.3% (2/6), respectively. No significant difference was found among them statistically (P > 0.05). The higher the differentiation grade of the esophageal squamous cell carcinoma, the more the express amount of pp60c-src (P < 0.005). The results suggested that the activation and express of c-src gene be associated with the initiation and development of the esophageal squamous cell carcinoma; the express amount of pp60c-src increased in the process and was associated with the differentiation of esophageal squamous cell carcinoma. PMID- 7490031 TI - [Co-carcinogenic effect of crocidolite plus benzo(a)pyrene on the lungs of rats]. AB - Epidemiological reports have indicated that occupational exposure to asbestos and smoking may greatly increase the incidence of lung carcinoma. In this study rats were injected intratracheally with crocidolite or benzo(a)pyrene [B(a)P] only, and crocidolite plus B(a)P in combination. The incidence of lung carcinomas in crocidolite group was 6.4% (3/47), in B(a)P group 10.4% (5/48) and crocidolite plus B(a)P group 46.3% (25/54). Besides, the survival curve and the mean survival time of rats with carcinoma dropped distinctly. Significant difference was shown among the crocidolite plus B(a)P group, crocidolite and B(a)P groups (P < 0.005). The result indicated the synergetic effect of crocidolite and B(a)P on the development of lung carcinoma in rats and the importance of giving up smoking in the people exposed to asbestos. A model of lung carcinoma in rat was successfully prepared for further study in experiment on treatment, prevention and mechanism. PMID- 7490033 TI - [Analysis of factors contributing to low birth weight in Sichuan Province. Cooperating Group for Birth Defects Monitoring]. AB - Data on low birth weight and some potential factors were obtained from an epidemiologic investigation of 111,676 births in Sichuan province during the period from Oct. 1986 to Sep. 1987. There were 8177 (73.22%) cases of low birth weight and 516 (79.89%) dead cases of low birth weight. The results suggest that the occurrence of low birth weight is affected by maternal age, fetal number and living level in rural areas. The incidence of low birth weight delivered by women aged 35 and over is 100.94%; it is 331.53% for twins and 265.96% for triplets. The incidence of low birth weight in rural areas (94.91%) is higher than that in urban areas (59.44%). PMID- 7490032 TI - [Some biochemical indexes in white rabbit's blood affected by acute high intensity microwave]. AB - Irradiation of white rabbits by 10, 50, 100 and 200 mW/cm2 microwave respectively can cause the disorder of protein metabolism, the abnormality of blood sugar, and the change of the activity of serum alpha-hydroxybutyrate dehydrogenase, lactate dehydrogenase, glutamic oxalacetic transaminase, glutamic pyruvic transaminase, acid phosphatase ect. These changes can be used as indexes in the evaluation of the effect of acute high intensity microwave exposure. The effect on the organism mainly depends on the intensity of exposure provided the dose of microwave remains the same. PMID- 7490034 TI - [Analyses of the prevalences for neural tube defects and cleft lip and palate in China from 1988 to 1991]. AB - This paper reports the monitoring results of two main categories of birth defects neural tube defects (NTD) and cleft of lip and palate (CLP) in China during the period of continuous monitoring (1988-1991). The method of hospital-based monitoring was adopted for data collection. From Jan. 1988 to Dec. 1991, a total of 2,750,588 births were monitored, and 28,168 cases of congenital malformations were identified. The prevalence of total birth defects was 102.4 per 10,000. 6885 cases of NTD were identified during this period, the prevalence of NTD being 25.0 per 10,000. The prevalence of NTD varied geographically; it declined gradually from the north to the south of China with the highest rate of NTD in the Taihang Mountain Regions. A total of 4548 cases of CLP were identified, and the provincial prevalence of CLP varied from 11.2 to 25.7 per 10000 with a mean of 16.5 per 10000. The prevalence of NTDs varied seasonally with the peak in winter (January). The sex ratio of NTD was 0.59, the prevalences in female births (32.1 per 10,000) being higher than that in male births (17.4 per 10,000) (P < 0.01). The sex ratio of CLP was 1.33. A significant difference was noted between the sex ratio of cleft palate (0.80) and that of the total cleft lip (1.46) (P < 0.01). The prevalence of NTD in rural areas (44.3 per 10,000) was 3 times as high as that in urban areas (14.4 per 10,000), but no difference in occurrence of CLP was observed between the urban and rural areas (P > 0.05). PMID- 7490035 TI - [Measurement of the reserve function of inspiratory muscle and its clinical significance]. AB - The principles of measuring inspiratory muscle tension-time index (TTim) and the ratio of the works of inspiration over the maximal works of inspiration (Wi/Wi(max)) were investigated and their formulae were deduced, i.e. TTim = (Pi x Ti)/(MIP x Ttot) and Wi/Wi(max) = (Pi x VT)/(MIP x IC). The importance of the inspiratory pressure and the maximal inspiratory pressure (MIP) measured at function residual capacity (FRC) level was emphasized. Both TTim and Wi/Wi(max) were measured in 35 normal subjects and 89 patients with chronic obstructive pulmonary disease (COPD). The results showed that normal value of TTim was 0.0253 +/- 0.0055 which corresponds to the normal value of the diaphragm tension-time index (TTdi = 0.02-0.03) reported by Bellemare. Patients with COPD had a mean TTim much higher than that of normals (P < 0.01). The works of inspiration (Wi) in patients with COPD increased, while the maximal works of inspiration (Wi(max)) declined, so Wi/Wi(max) became significantly greater than that of normals (P < 0.01). The results also showed that there was a linear relationship between Pi/Pimax and TTim or Wi/Wimax (r = 0.7891, 0.9738, 0.6459, 0.9327, P < 0.01). Therefore, we suggest that both TTim and Wi/Wimax can be used as clinical indices to reflect the reserve function of inspiratory muscles. PMID- 7490036 TI - [Immunohistochemical study of 38 cases of primary ocular adnexa malignant lymphomas]. AB - Thirty-eight cases of primary ocular adnexa malignant lymphomas were studied with immunohistochemical techniques. Positive results were noted in 14 cases of kappa light chain staining, 8 cases of lambda staining and 1 case of centroblastic lymphoma lambda staining. In 35 cases of L26 staining, positive results were observed, which showed that 92.1% of the lymphomas were of B cell origin whereas those of T cell origin were rare. The value of immunohistochemical methods in diagnosis and differential diagnosis as well as their significance for clinical treatment and prognosis were commented. PMID- 7490037 TI - [A 3-year follow-up study of 50 children with acute lymphoblastic leukemia]. AB - Fifty children with acute lymphoblastic leukemia were followed up for an average period of 3 3/12 years. The results showed that 19 (38%) cases were given up after diagnosis. Eight patients only received treatment irregularly for 1 year; among them, 2 discontinued therapy by themselves for 2-3 years and survived free of events, and 6 were lost, Twenty-three received treatment regularly; among them, 3 died of infection, 11 were in continuously complete remission and 9 experienced relapse. The total disease-free survival rate for mean 3 3/12 years is 42% (13/31), exclusive of the 19 given up cases. Statistical analysis indicates a higher relapse rate in those cases with lower initial glucocorticoid receptors (GCR) of peripheral blood lymphocytes. PMID- 7490038 TI - [Simultaneous assay of Ca(2+)-ATPase and Na+, K(+)-ATPase activities of hemolysate by malachite green colorimetric method]. AB - Activities of Ca(2+)-ATPase and Na+, K(+)-ATPase of hemolysate were simultaneously determined by a colorimetric method with malachite green dye. In the presence of Ca2+, EGTA or ouabain, inorganic phosphate released from ATP by hypotonic saponin-treated red cell lysate was used to estimate the activities of Ca(2+)-ATPase and Na2+, K(+)-ATPase. The method is simple, sensitive and stable for at least 4 h. The effects of concentration of saponin and Ca2+, the storage of hemolysate, and the activator and inhibitor of the enzymes on activity were discussed. PMID- 7490039 TI - [Study on determination of gentamycin in serum by enhanced chemiluminesent immunoassay based on immobilized antibody and biotin-avidin system]. AB - Gentamycin was determined by competition with biotin labeled gentamycin in the presence of an immobilized antibody to gentamycin. Microcrystalline cellulose was first coupled with sheep anti-rabbit antibody, then the anti-gentamycin antibody was conjugated and the competition reaction conducted. After separation, HRP avidin was added to the precipitation and the amount of HRP conjugate determined by enhanced chemiluminescene. The limit of detection for gentamycin in serum was 63-78 ng/tube and the average recovery of gentamycin 103%. The correlation coefficient of the log-log regression equation. Magnitude of r was 0.991 +/- 0.006. PMID- 7490040 TI - [Preconcentration of trace chromium in natural water with activated carbon and its determination by colorimetry]. AB - The adsorption and desorption behavior of chromium was studied. Trace chromium in water could be adsorbed on activated carbon at pH 9-10, then eluted with 0.5% sulfuric acid and determined by colorimetry. The content of chromium could be concentrated by 100 times. For sample, the coefficient of variation of 8 parallel determinations was 7.7%. The proposed method has been applied to the determination of trace chromium in natural waters with the recovery of 71.4% 90.0%, and with the detection limit of 0.04 microgram/L. PMID- 7490043 TI - Trends in female sexual behaviour and sexually transmitted diseases in London, 1982-1992. AB - OBJECTIVE: To measure changes in female sexual behaviour, including condom use, and their relationship with the incidence of sexually transmitted and other genital diseases in women during the decade 1982-92. DESIGN: A prospective series of cross-sectional surveys of sexual behaviour reported by a standardised self administered questionnaire in new patients who presented for screening and diagnosis. SETTING: A genitourinary medicine clinic in West London. SUBJECTS: 4089 consecutive newly attending patients who completed sexual behaviour questionnaires during 1982, 1987, 1989 and 1992. MAIN OUTCOME MEASURES: Trends in socio-demographic status, sexual behaviour, condom-use, sexually transmitted diseases and other genital infections diagnosed by routine clinical and laboratory methods. RESULTS: Women reported significantly increasing condom use (from 3.6% to 20.7%) and decreasing oral contraception (from 51.2% to 40.1%), but the proportion who used no contraception (23.6% to 24.7%) and the proportion who had never been pregnant (58.3% to 59.9%) remained similar. Numbers of sexual partners in the preceding year decreased (p < 0.001) and an increasing proportion of women practised oral intercourse (p < 0.001). During the same period, there was a progressive decline (p < 0.001) in the incidence of gonorrhoea, chlamydial infection and trichomoniasis by approximately two-thirds. However, the incidence of vaginal candidosis (p < 0.001), bacterial vaginosis (p < 0.001) and genital warts (p < 0.01) increased. CONCLUSIONS: Increasing use of condoms for vaginal intercourse with both regular and non-regular partners has been associated with a decrease in the incidence of gonorrhoea, chlamydial infection and trichomoniasis. There was also an increase in the practice of fellatio and a change in the spectrum of STD and other genital infections with little net reduction in morbidity. HIV infection showed no evidence of heterosexual spread. PMID- 7490042 TI - Open lung biopsy for investigation of acute respiratory episodes in patients with HIV infection and AIDS. AB - BACKGROUND: Open lung biopsy (OLB) is rarely necessary for investigation of HIV positive patients with acute respiratory episodes because of the high yield from fibreoptic bronchoscopy with bronchoalveolar lavage (BAL). METHODS: A retrospective review of OLB in HIV positive patients admitted to a specialist inpatient unit with acute respiratory symptoms was carried out in order to define clinical indications, diagnostic yield, impact on management, complications and outcome. RESULTS: OLB was performed in 23 patients; 21 had undergone one or more bronchoscopies with BAL (5 also had negative results from transbronchial biopsy). Indications for OLB were: Group A, 15 patients thought clinically to have pneumocystis pneumonia but not responding to treatment; Group B, 4 patients with focal chest radiographic abnormalities; Group C, 4 patients with diffuse radiographic abnormalities and miscellaneous conditions. Preoperative PaO2 (on air) ranged from 4.4 to 14.5 (mean = 9.5) kPa. The results of OLB were in Group A 5 patients had non specific interstitial pneumonitis (NIP), 1 also had Kaposi's sarcoma, 4 had pneumocystis pneumonia (1 also had bronchiolitis obliterans organising pneumonia [BOOP]), 3 had Kaposi's sarcoma and 1 had BOOP and emphysema, 1 had pulmonary infarction and no infection and 1 had normal lung tissue. In Group B diagnoses were NIP, B cell lymphoma, occult alveolar haemorrhage and Pseudomonas aeruginosa pneumonia with BOOP; In Group C 2 patients had NIP and 2 had pneumocystis pneumonia (1 also had cytomegalovirus pneumonitis). All patients survived surgery and none required mechanical ventilation. OLB results significantly affected management; in Group A inappropriate treatment was discontinued in 11 patients found not to have pneumocystis pneumonia, and alternative therapy was begun in the 4 with pneumocystis and in Groups B and C 6 patients began specific therapy; unnecessary therapy was avoided in one and antimicrobial treatment was modified in one. CONCLUSIONS: Open lung biopsy in HIV positive patients with focal and diffuse radiographic abnormalities has a high diagnostic yield and low morbidity. This investigation should be considered in those with acute respiratory episodes and negative results from bronchoscopic investigations or who have contra-indications to this procedure. PMID- 7490041 TI - Similar serological response to conventional therapy for syphilis among HIV positive and HIV-negative women. AB - OBJECTIVES: To compare characteristics of syphilis serological reactivity in HIV positive (+) and HIV negative (-) female sex workers, as well as the serological response to therapy after treatment with intramuscular benzathine penicillin, 2.4 million U weekly, for three consecutive weeks. METHODS: Rapid plasma reagin (RPR) and Treponema pallidum haemagglutination assay (TPHA) results of 72 HIV-positive and 121 HIV-negative women reactive in both tests were assessed. The response to therapy was prospectively monitored with quantitative RPR serology in 47 HIV positive and 73 HIV-negative patients. Cumulative probabilities of becoming nonreactive by RPR were compared at six months, one and two years after therapy. RESULTS: At enrolment, the geometric mean titres of RPR and TPHA were lower in HIV-positive patients (RPR, 1:2.6) than in HIV-negative patients (RPR, 1:3.8; p < 0.01). The evolution over time of RPR titres was similar among HIV-positive patients as compared to HIV-negative patients. Among patients with an initial RPR titre of < 1:8, 53% of HIV-positive and 44% of HIV-negative patients became RPR negative two years after therapy. Among patients with an RPR titre of 1:8 or greater at enrolment, 83% of HIV-positive and 90% of HIV-negative patients had reached at least a fourfold decline of RPR titres two years after therapy. CONCLUSIONS: Syphilis serology findings (both RPR and TPHA) may be altered in the presence of HIV infection, but the serological response to therapy was similar in HIV-positive and HIV-negative patients. PMID- 7490044 TI - Heterosexual relationships and condom-use in the spread of sexually transmitted diseases to women. AB - OBJECTIVES: To examine the effect of patient-defined non-regular heterosexual relationships on the incidence of sexually transmitted diseases and other genital infections in women and the role of condom use in the prevention of their spread. DESIGN: A cross-sectional study of sexual behaviour reported by a standardised self-administered questionnaire in new patients who presented for screening and diagnosis. SETTING: A genitourinary medicine clinic in West London. SUBJECTS: 938 consecutive newly attending women who completed a sexual behaviour questionnaire in 1992. MAIN OUTCOME MEASURES: Variables relating to socio-demographic status, sexual behaviour, condom use, sexually transmitted diseases and other genital infections stratified by the reporting of non-regular partners. RESULTS: We found that women who reported non-regular sexual partners were more likely to be single (p = 0.0001), white (p < 0.0001), have had coitarche before 17 years of age (p = 0.003) and many more sexual partners both in the last year and in their life-time (p < 0.0001) and were more likely to practise fellatio (p < 0.0001), anal penetration (p = 0.004) and to be smokers (p < 0.0001). Paradoxically, the incidence of sexually transmitted diseases and other genital infections was no higher in this group than in the group of women who did not have non-regular partners. Increasing condom use with regular partners correlated with decreasing incidence of gonorrhoea (p < 0.001), chlamydial infection (p < 0.01) and trichomoniasis (p < 0.02), but increasing condom use with non-regular partners did not show this trend. CONCLUSIONS: Regular heterosexual partners play the major role in transmission of bacterial sexually transmitted diseases to women. This is significantly influenced by use of condoms. PMID- 7490046 TI - Genotyping of Chlamydia trachomatis serovars derived from heterosexual partners and a detailed genomic analysis of serovar F. AB - OBJECTIVE: To investigate C trachomatis serovars in contact-traced heterosexual partners. METHODS: Urogenital Chlamydia trachomatis isolates (n = 112) derived from 35 heterosexual patients (index patients) and their 37 chlamydia positive partners (contact patients) were differentiated into serovars by genotyping with restriction fragment length polymorphism (RFLP) analysis of the PCR amplified omp1 gene. In order to investigate whether different strains within the frequently prevalent serovar F were transmitted, two pairs of serovar F (n = 4) were further analysed by genomic DNA fingerprinting with arbitrary primer PCRs (AP-PCRs). RESULTS: Identical C trachomatis serovars were found in 31 of the 35 pairs, serovars E, F, D, and G being most prevalent. In the remaining four pairs different serovars (either D, E, F or G) were found between the index and the contact patients. By AP-PCR analysis the strains of serovar F were found to be identical between the index and the contact patients, but were different between the two pairs in all AP-PCRs used. CONCLUSION: A majority of heterosexual partners, once traced positive for C trachomatis infections, are infected with identical serovars. Identical strains of serovar F found in partners as found by DNA fingerprinting confirms the sexual transmission of C trachomatis. PMID- 7490045 TI - Prevalence of Chlamydia trachomatis in women attending a family planning clinic in Papua New Guinea. AB - OBJECTIVE: To determine the prevalence of Chlamydia trachomatis infection in women attending a family planning clinic in Papua New Guinea, in the period between April and June 1991. SETTING: The outpatient department of Obstetrics and Gynaecology of Port Moresby General Hospital, Port Moresby, Papua New Guinea, the departments of Dermato-Venereology and Clinical Microbiology of the Erasmus University, Rotterdam, The Netherlands and the National Institute of Public Health and Environmental Protection, Bilthoven, The Netherlands. PATIENTS: A total of 254 consecutive women who attended the family planning clinic at Port Moresby General Hospital, Papua New Guinea were enrolled into this study. METHODS: Cervical infections with C trachomatis were diagnosed using the direct immunofluorescent assay (DFA) and the polymerase chain reaction (PCR). Serum IgM and IgG antibodies directed against C trachomatis were detected using the enzyme linked fluorescent assay (ELFA). RESULTS: The prevalence of C trachomatis was 14.6% using the PCR, 9.1% using the DFA and 17.3% when the results of the PCR and the DFA were combined. An elevated IgM titre was observed in 14.2% of the women, whereas 44.1% had an elevated IgG titre. The titres of IgM or IgG were significantly higher in women who were positive using the PCR or the DFA than in those who were negative in both the PCR and the DFA (p = 0.032 and p = 0.0046, respectively). CONCLUSION: Cervical infection by C trachomatis can be considered a major health problem in at least the studied population in Papua New Guinea. The prevalence of C trachomatis infection is at least comparable with that in groups with a high prevalence in industrialized countries. Effective screening and treatment programmes are imperative to combat this problem. PMID- 7490047 TI - Chronic vulvovaginal candidiasis: characteristics of women with Candida albicans, C glabrata and no candida. AB - INTRODUCTION: Although as many as 5% of all women complain of chronic vulvovaginitis, little is known about these women. They may often be misdiagnosed and the role of vaginal yeast culture in diagnosing vulvovaginal candidiasis (VVC) among them has not been clearly defined. METHODS: To address these deficiencies, we tabulated initial diagnoses among new patients and conducted a medical record-based, unmatched case-control study among women reporting a history of chronic vulvovaginitis (four or more episodes in the past year) at a vulvovaginitis specialty clinic. Clinical presentation and medical history were compared for women who had a positive vaginal yeast culture for either Candida albicans or C glabrata, or who had a negative culture. RESULTS: One-third of the women had no apparent vulvovaginal disease at their initial visit. All women reported similar symptoms, except for an increased prevalence of painful sexual intercourse in women with C albicans (chi 2 p = 0.014 versus women with C glabrata and p < 0.001 versus women with no candida). Women with C glabrata were more likely to be non-white (chi 2 p = 0.071 compared with women with C albicans) and to report an underlying medical condition (chi 2 p < or = 0.001 versus both women with C albicans and women with no candida). Physical examination was normal only in women with no candida. C albicans cases were more likely to have positive potassium hydroxide microscopy (chi 2 p = 0.016) and a pH < or = 4.5 (chi 2 p = 0.011) than were C glabrata cases. CONCLUSIONS: These results suggest that reliance on symptoms and signs alone will result in significant misdiagnosis of chronic vulvovaginitis. Among women with VVC, subtle differences in clinical presentation do not reliably distinguish women with C albicans from those with C glabrata. Our study also indicates that vaginal yeast cultures, while not necessary for every patient, are valuable in confirming negative diagnoses, detecting microscopy false-negatives, and identifying non-C albicans isolates. PMID- 7490048 TI - Disseminated cutaneous Mycobacterium tuberculosis infection in a patient with AIDS. AB - Individuals infected with the human immunodeficiency virus (HIV) are at an increased risk of both pulmonary and extrapulmonary tuberculosis. Disseminated cutaneous tuberculosis is rare, but has been reported in four HIV-positive patients, all of whom also had pulmonary infection. In this report we describe an HIV-infected patient with a febrile illness and an abnormal chest radiograph who developed widespread cutaneous tuberculous pustules following a lymph node biopsy on the previous day. PMID- 7490050 TI - A publisher, a bobbin-boy and the Society. PMID- 7490049 TI - Vulvitis plasmacellularis: two new cases. AB - Vulvitis chronica plasmacellularis or Zoon's vulvitis is a rare benign circumscribed inflammation of the vulvar mucosa. It is found in women ranging in age from 26 to 70 years. Shiny, macular erythematous lesions, which are irregular in shape and sharply marginated are usually observed. The histologic findings show chronic subepithelial dense inflammation composed largely of plasma cells. We here report two cases of vulvitis plasmacellularis with typical clinical manifestations, courses and histopathologic findings. PMID- 7490051 TI - HIV infection and maternal outcome of pregnancy in Mozambican women: a case control study. PMID- 7490052 TI - Antiretroviral monotherapies and serum HIV-1 dynamics. PMID- 7490053 TI - Failure of itraconazole to prevent Enterocytozoon bieneusi infection. PMID- 7490054 TI - Chlamydia trachomatis in gynaecological infections in Luanda, Angola. PMID- 7490055 TI - Sensitivity of a commercial polymerase chain reaction for different serovars of Chlamydia trachomatis present at low titre in clinical samples. PMID- 7490056 TI - Urethral flora in adolescent boys. PMID- 7490058 TI - Perihepatitis in women with salpingitis--an under-diagnosed clinical entity? PMID- 7490057 TI - Screening for asymptomatic Chlamydia trachomatis infection in male students by examination of first catch urine. PMID- 7490060 TI - Patients' awareness of changes in the Association of British Insurers' guidelines on HIV testing. PMID- 7490061 TI - How much sexually transmitted disease is not seen by genitourinary physicians? PMID- 7490059 TI - Importation into the UK of a strain of Neisseria gonorrhoeae resistant to penicillin, ciprofloxacin and tetracycline. PMID- 7490062 TI - Child sexual abuse--the interface with genitourinary medicine. PMID- 7490063 TI - Family planning in genitourinary medicine. PMID- 7490064 TI - Rectal gonorrhoea as an independent risk factor for HIV infection in homosexual males. PMID- 7490065 TI - A case cluster of possible tissue invasive gonorrhoea. PMID- 7490066 TI - Pneumococcal vaccine and HIV infection. PMID- 7490067 TI - Pseudomonas aeruginosa infections and HIV. PMID- 7490068 TI - Carcinoma of the penis: A cluster of cases in young men. PMID- 7490069 TI - Identification of four novel human genes amplified and overexpressed in breast carcinoma and localized to the q11-q21.3 region of chromosome 17. AB - We have performed differential screening of a human metastatic lymph lymph node cDNA library to identify genes possibly involved during breast cancer progression. We have identified four novel genes overexpressed in malignant tiddues. They were all located on the long arm of chromosome 17, in loci located between q11 and q21.3, a region known to contain the c-erbB-2 oncogene and the BRCA1 breast carcinomas, and overexpression of three of them was dependent on gene amplification in breast cancer cell lines. These findings further support the concept that human chromosome 17 specifically carries genes possibly involved in breast cancer progression. PMID- 7490070 TI - A comparative transcriptional map of a region of 250 kb on the human and mouse X chromosome between the G6PD and the FLN1 genes. AB - The transcriptional organization of the region of the mouse X chromosome between the G6pd and the Fln1 genes was studied in detail, and it was compared with the syntenic region of the human chromosome. A cosmid contig of 250 kb was constructed by screening mouse cosmid libraries with probes for human genes and with whole cosmids. Overlapping cosmids were aligned by comparing EcoRI and rare cutter restriction enzyme digestions. The gene order and the orientation of transcription were determined by hybridization with fragments from the 5' and 3' moieties of each cDNA. Our work demonstrates that all of the new genes identified in human are present in the mouse. The size of the region, 250 kb, is also very similar, as are gene order and gene organization: the transcriptional organization in "domains" described in human is found to be identical in the mouse. The major difference detected is the much lower content in rare-cutter restriction sites, which is related to the lower G+C and CpG content of mouse DNA. The very high conservation that we have described suggests that a potent selective pressure has contributed to such conservation of gene organization. PMID- 7490071 TI - A YAC contig spanning the dominant retinitis pigmentosa locus (RP9) on chromosome 7p. AB - The dominant retinitis pigmentosa locus RP9 has previously been localized to 7p13 p15, in the interval D7S526-D7S484. We now report refinement of the locus to the interval D7S795-D7S484 and a YAC contig of approximately 4.8 Mb spanning this region and extending both distally and proximally from it. The contig was constructed by STS content mapping and physically orders 29 STSs in 28 YAC clones. The order of polymorphic markers in the contig is consistent with a genetic map that has been assembled using haplotype data from the CEPH pedigrees. This contig will provide a primary resource for the construction of a transcriptional map of this region and for the identification of the defective gene causing this form of adRP. PMID- 7490072 TI - The FSHD-associated repeat, D4Z4, is a member of a dispersed family of homeobox containing repeats, subsets of which are clustered on the short arms of the acrocentric chromosomes. AB - Facioscapulohumeral muscular dystrophy (FSHD) is an autosomal dominant neuromuscular disorder that maps to human chromosome 4q35. FSHD is tightly linked to a polymorphic 3.3-kb tandem repeat locus, D4Z4. D4Z4 is a complex repeat: it contains a novel homeobox sequence and two other repetitive sequence motifs. In most sporadic FSHD cases, a specific DNA rearrangement, deletion of copies of the repeat at D4Z4, is associated with development of the disease. However, no expressed sequences from D4Z4 have been identified. We have previously shown that there are other loci similar to D4Z4 within the genome. In this paper we describe the isolation of two YAC clones that map to chromosome 14 and that contain multiple copies of a D4Z4-like repeat. Isolation of cDNA clones that map to the acrocentric chromosomes and Southern blot analysis of somatic cell hybrids show that there are similar loci on all of the acrocentric chromosomes. D4Z4 is a member of a complex repeat family, and PCR analysis of somatic cell hybrids shows an organization into distinct subfamilies. The implications of this work in relation to the molecular mechanism of FSHD pathogenesis is discussed. We propose the name 3.3-kb repeat for this family of repetitive sequence elements. PMID- 7490073 TI - Type 1 diabetes and the control of dexamethazone-induced apoptosis in mice maps to the same region on chromosome 6. AB - Quantitative trait loci mapping was used to identify the chromosomal location of genes that contribute to increase the resistance to apoptosis induced in immature CD4+8+ thymocytes. An F2 intercross of the nonobese diabetic (NOD) mouse (displaying an apoptosis-resistance phenotype) and the C57BL/6 mouse (displaying a nonresistance phenotype) was phenotypically analyzed and genotyped for 32 murine microsatellite polymorphisms. Maximum likelihood methods identified a region on the distal part of chromosome 6 that is linked to dexamethazone-induced apoptosis (lod score = 3.46) and accounts for 14% of the phenotypic variation. This chromosomal region contains the diabetes susceptibility locus Idd6, suggesting that the apoptosis-resistance phenotype constitutes a pathogenesis factor in IDDM of NOD mice. PMID- 7490074 TI - Structure and chromosomal localization of the mouse neurocan gene. AB - Cosmid clones containing the mouse neurocan gene were isolated from a genomic library using rat neurocan cDNA fragments as probe. The murine gene has a size of approximately 25 kb and contains the coding sequence for the mRNA on 15 exons. The exon-intron structure reflected the structural organization of neurocan, which is a multidomain protein belonging to the aggrecan/versican proteoglycan family. All introns between conserved modular protein domains are phase I introns. Primer extension experiments indicate a transcriptional start point 28 bases downstream of a consensus TATA sequence. Further analysis of 1 kb of 5' flanking sequence revealed in addition to AP1, AP2, and SP1 consensus binding sites multiple E-box elements and a glucocorticoid responsive element. Single strand conformation polymorphism was used to map neurocan to chromosome 8 between the microsatellite markers D8Mit29 and D8Mit78. Among mouse mutants that have been mapped around this region are the three allelic neurological diseases tottering, leaner, and rolling. The multidomain structure and the preferential expression of neurocan in the brain suggest a potential involvement in these diseases. PMID- 7490075 TI - Structure of the gene for the catalytic subunit of human DNA polymerase delta (POLD1). AB - We have isolated genomic DNA clones covering the gene for human DNA polymerase delta catalytic subunit (POLD1) and its 5' flanking sequence. This gene is divided into 27 exons and is distributed over at least 32 kb of DNA. The exons and most of the introns are relatively small. The sizes of the exons range from 55 to 201 bp. Seven introns are smaller than 100 bp. Intron 1 is the largest intron, with a size of greater than 10 kb. All of the intron-exon junctions match well with the reported consensus sequences. Multiple copies of the Alu repetitive sequence and the variable number of tandem repeats were found in several introns. Transcription of POLD1 appears to initiate at multiple sites. The major start site was 53 nucleotides upstream of the ATG start codon. The sequence of the promoter and upstream DNA is G+C rich and does not contain a TATA sequence. Several potential transcription factor-binding sites, including the AP2-, CTF-, Ets1-, GCF-, MBF-1-, NF-E1-, and Sp1-binding sites, were found in this region. A 1.8-kb pol delta promoter DNA directed the expression of a luciferase reporter gene when transfected into HeLa cells. PMID- 7490076 TI - A contiguous clone map over 3 Mb on the long arm of chromosome 11 across a balanced translocation associated with schizophrenia. AB - Forty-nine clones derived by microdissection of a schizophrenia-associated t(1;11)(q42.1;q14.3) breakpoint region have been assigned by somatic cell hybrid mapping to seven discrete intervals on the long arm of human chromosome 11. Eleven of the clones were shown to map to a small region immediately distal to the translocation breakpoint on 11q. A 3-Mb contiguous clone map of this region was established by isolation of corresponding YAC recombinants. The contig was oriented and shown to traverse the translocation breakpoint by FISH and microsatellite marker analysis. This contig will facilitate the isolation of candidate sequences whose expression may be affected by the translocation. PMID- 7490077 TI - Gene structure and amino acid sequence of the human cone photoreceptor cGMP phosphodiesterase alpha' subunit (PDEA2) and its chromosomal localization to 10q24. AB - The genomic organization and nucleotide structure of the human cone photoreceptor cGMP phosphodiesterase alpha'-subunit (alpha'-PDE) gene (PDEA2) as well as its chromosomal localization have been determined. This gene, which spans about 48 kb, consists of 22 exons and codes for an 858-amino-acid protein. The alpha'-PDE gene maps to human chromosome 10q24. Its coding region shows about 90% nucleotide identity and 93% amino acid identity with the corresponding region of the bovine gene. The intron-exon organization of the genes encoding human cone alpha'-PDE and rod beta-PDE are very similar, suggesting that these proteins have a close phylogenetic relationship and probably a common origin. PMID- 7490078 TI - Efficient isolation of novel mouse genes differentially expressed in early postimplantation embryos. AB - Most genes with regulatory functions in embryogenesis are expressed in highly specific patterns, suggesting that expression patterns can serve as criteria to define potential candidates for developmentally relevant genes. To isolate such genes, we selected and partially sequenced 80 cDNA clones from a 10.5-day mouse embryo library. Forty-one clones that represented novel mouse genes were analyzed for expression in embryos of the same stage by whole-mount in situ hybridization. A high proportion (24%) of these genes, including a homologue of the Drosophila Delta gene, were expressed in specific spatially restricted patterns, suggesting that selection based on expression patterns is a useful strategy to isolate novel genes that may play pivotal roles in mammalian development. PMID- 7490079 TI - Molecular cloning of the human homolog of a striatum-enriched phosphatase (STEP) gene and chromosomal mapping of the human and murine loci. AB - A gene for a protein tyrosine phosphatase (PTPase) was isolated from a human fetal brain cDNA library by PCR amplification. Sequence analysis revealed that the PTPase has a single phosphatase catalytic domain located at the C-terminus that includes the highly conserved amino acid domain [I/V]HCXAGXXR[S/T]GX[F/Y] found in all tyrosine phosphatases. Two proline-rich regions located at the N terminus may contain putative Src homology domain 3 (SH3) binding motifs. Comparison of the PTPase with a previously cloned striatum enriched phosphatase (STEP) from rat and from mouse exhibited a high degree of identity (approximately 85-90%) at both the nucleotide and the amino acid levels, indicating that the human PTPase is the homolog of the rat and murine STEP gene. By using a combination of somatic cell hybrid analysis and fluorescence in situ hybridization, we have mapped the human STEP locus to chromosome 11p15.2-p15.1 and the murine STEP gene to chromosome 7B3-B5. These are two regions of known conserved synteny, providing further evidence that the human STEP is a true homolog of the murine STEP gene. Candidate disease genes in the vicinity include Usher syndrome type 1C in human and a mouse mutant locus, twister (twt). PMID- 7490080 TI - Mammalian mitochondrial intermediate peptidase: structure/function analysis of a new homologue from Schizophyllum commune and relationship to thimet oligopeptidases. AB - Mitochondrial intermediate peptidase (MIP) is a component of the mitochondrial protein import machinery required for maturation of nuclear-encoded precursor proteins targeted to the mitochondrial matrix or inner membrane. We previously characterized this enzyme in rat (RMIP) and Saccharomyces cerevisiae (YMIP) and showed that MIP activity is essential for mitochondrial function in yeast. We have now defined the structure of a new MIP homologue (SMIP) from the basidiomycete fungus Schizophyllum commune. SMIP includes 4 exons of 523, 486, 660, and 629 bp separated by 3 short introns. The predicted SMIP, YMIP, and RMIP sequences share 31-37% identity and 54-57% similarity over 700 amino acids. When SMIP and RMIP were expressed in a yeast mip1 delta mutant, they were both able to rescue the respiratory-deficient phenotype caused by genetic inactivation of YMIP, indicating that the function of this enzyme is conserved in eukaryotes. Moreover, the MIP sequences show 20-24% identity and 40-47% similarity to a family of oligopeptidases from bacteria, yeast, and mammals. MIP and these proteins are characterized by a highly conserved motif, F-H-E-X-G-H-(X)2-H-(X)12 G-(X)5-D-(X)2-E-X-P-S-(X)3-E-X, centered around a zinc-binding site and appear to represent a new family of genes associated with proteolytic processing in the mitochondrial and cytosolic compartments. PMID- 7490081 TI - Molecular structure of the Menkes disease gene (ATP7A). AB - We report a detailed molecular analysis of the genomic structure of the Menkes disease gene (MNK; ATP7A). There are 23 exons in ATP7A covering a genomic region of approximately 140 kb. The size of the individual coding exons varies between 77 and 726 bp, and introns vary in size between 196 bp and approximately 60 kb. All of the splice sites obey the consensus GT-AG rule except the splice donor of intron 9, which is GC instead of GT. The exon following this rare splice donor variant is alternatively spliced. A PGAM pseudogene and two highly polymorphic CA repeats map to introns within the gene. The structure is very similar to that of the closely related Wilson disease gene (WND; ATP7B). From exon 5 (exon 3 in ATP7B) to the end, all of the splice sites occur at exactly the same nucleotide positions as in the WND gene, except for the boundary between exons 17 and 18 (exons 15 and 16 in ATP7B) and a single codon difference at the boundary between exons 4 and 5 of the MNK gene (exons 2 and 3 in ATP7B). In contrast to the WND gene, in which the first four of six metal binding domains are contained in 1 exon, metal binding domains 1 to 4 are divided over 3 exons. The striking similarity of the MNK and WND genes at the genomic level is consistent with their relatively recent divergence from a common ancestral gene. PMID- 7490082 TI - Cloning of the cDNA for the human ATP synthase OSCP subunit (ATP5O) by exon trapping and mapping to chromosome 21q22.1-q22.2. AB - Exon trapping was used to clone portions of potential genes from human chromosome 21. One trapped sequence showed striking homology with the bovine and rat ATP synthase OSCP (oligomycin sensitivity conferring protein) subunit. We subsequently cloned the full-length human ATP synthase OSCP cDNA (GDB/HGMW approved name ATP50) from infant brain and muscle libraries and determined its nucleotide and deduced amino acid sequence (EMBL/GenBank Accession No. X83218). The encoded polypeptide contains 213 amino acids, with more than 80% identity to bovine and murine ATPase OSCP subunits and over 35% identity to Saccharomyces cerevisiae and sweet potato sequences. The human ATP5O gene is located at 21q22.1 q22.2, just proximal to D21S17, in YACs 860G11 and 838C7 of the Chumakov et al. (Nature 359:380, 1992) YAC contig. The gene is expressed in all human tissues examined, most strongly in muscle and heart. This ATP5O subunit is a key structural component of the stalk of the mitochondrial respiratory chain F1F0-ATP synthase and as such may contribute in a gene dosage-dependent manner to the phenotype of Down syndrome (trisomy 21). PMID- 7490083 TI - Genomic organization of the human gene (CA5) and pseudogene for mitochondrial carbonic anhydrase V and their localization to chromosomes 16q and 16p. AB - Carbonic anhydrase V (CA V) is expressed in mitochondrial matrix in liver and several other tissues. It is of interest for its putative roles in providing bicarbonate to carbamoyl phosphate synthetase for ureagenesis and to pyruvate carboxylase for gluconeogenesis and its possible importance in explaining certain inherited metabolic disorders with hyperammonemia and hypoglycemia. Following the recent characterization of the cDNA for human CA V, we report the isolation of the human gene from two lambda genomic libraries and its characterization. The CA V gene (CA5) is approximately 50 kb long and contains 7 exons and 6 introns. The exon-intron boundaries are found in positions identical to those determined for the previously described CA II, CA III, and CA VII genes. Like the CA VII gene, CA5 does not contain typical TATA and CAAT promoter elements in the 5' flanking region but does contain a TTTAA sequence 147 nucleotides upstream of the initiation codon. CA5 also contains a 12-bp GT-rich segment beginning 13 bp downstream of the polyadenylation signal in the 3' untranslated region of exon 7. FISH analysis allowed CA5 to be assigned to chromosome 16q24.3. An unprocessed pseudogene containing sequence homologous to exons 3-7 and introns 3-6 was also isolated and was assigned by FISH analysis to chromosome 16p11.2-p12. PMID- 7490084 TI - Characterization of a novel zinc finger gene (ZNF165) mapping to 6p21 that is expressed specifically in testis. AB - Clone ZF388 was identified during the course of expressed sequence tag analysis of human adult testis cDNAs as being a member of the C2H2 zinc finger gene family. Northern blot analysis of a range of 16 different human tissues has shown that clone ZF388 detects a single 2.2-kb transcript and that the expression of this mRNA species is strictly specific to the testis. Sequence analysis of ZF388 and other clones isolated by rescreening a testis cDNA library has defined a transcript with an extensive N-terminal acidic domain containing 17% aspartic and glutamic acid residues and a C-terminal domain composed of five zinc finger motifs linked through the highly conserved sequence TGE-KPYE. This gene has been designated ZNF165. Analysis of somatic cell hybrids containing single human chromosomes shows that ZNF165 maps to chromosome 6. Fluorescence in situ hybridization of cDNA insert probes to male metaphase spreads shows that the ZNF165 transcript maps to 6p21 close to the human MHC region. Human 6p21 is homologous with the distal inversion of the mouse t-complex, which has been shown to contain a cluster of genes expressed specifically in the testis. PMID- 7490085 TI - The genomic organization of the human transcription factor 3 (TFE3) gene. AB - We have determined the exon-intron structure of the human TFE3 gene located on Xp11.22-23. By designing PCR primers, we were able to amplify various segments of the TFE3 genomic region, thus establishing that this gene is composed of seven exons, the first six of which are small (from 56 to 159 nt). The 5' UT region is contained entirely in the first exon, while the 3' UT region is contained in the seventh exon. The comparison of the genomic and the published cDNA versions revealed that the deduced amino acid sequence of TFE3 in the C-terminus region is 125 amino acids shorter than previously reported. This eliminates most of the putative proline- and arginine-rich domain and makes the human sequence more similar to its mouse homolog. The activation domain at the N-terminus is contained in exon 2, as has been described for the mouse. The basic helix-loop helix (BHLH) motif is spread over exons 4 to 6, while the leucine zipper (LZ) is almost all contained in the last portion of exon 6. This split BHLH is different from other BHLH-LZ genes whose genomic structures have been determined up to now. PMID- 7490086 TI - Structure and chromosomal localization of the human stromal cell-derived factor 1 (SDF1) gene. AB - Stromal cell-derived factors 1 alpha and 1 beta are small cytokines belonging to the intercrine CXC subfamily and originally isolated from a murine bone-marrow stroma cell line by the signal sequence trap method. cDNA and genomic clones of human SDF1 alpha and SDF1 beta (SDF1A and SDF1B) were isolated and characterized. cDNAs of SDF1 alpha and SDF1 beta encode proteins of 89 and 93 amino acids, respectively. SDF1 alpha and SDF1 beta sequences are more than 92% identical to those of the human counterparts. The genomic structure of the SDF1 gene revealed that human SDF1 alpha and SDF1 beta are encoded by a single gene and arise by alternative splicing. SDF1 alpha and SDF1 beta are encoded by 3 and 4 exons, respectively. Ubiquitous expression of the SDF1 gene, except in blood cells, was consistent with the presence of the GC-rich sequence in the 5'-flanking region of the SDF1 gene, as is often the case in the "housekeeping" genes. Although genes encoding other members of the intercrine family are localized on chromosome 4q or 17q, the human SDF1 gene was mapped to chromosome 10q by fluorescence in situ hybridization. Strong evolutionary conservation and unique chromosomal localization of the SDF1 gene suggest that SDF1 alpha and SDF1 beta may have important functions distinct from those of other members of the intercrine family. PMID- 7490087 TI - Structure of the murine MPTP-PEST gene: genomic organization and chromosomal mapping. AB - Protein tyrosine phosphatases comprise a large family of enzymes that are involved in the control of cellular tyrosine phosphorylation. We have used lambda phage analysis to elucidate the complete genomic structure of an intracellular member of this family, the murine MPTP-PEST gene. Eight overlapping lambda phage clones representing the MPTP-PEST locus were isolated from a 129/sv mouse genomic library. The gene spans over 90 kb of the mouse genome and is composed of 18 exons, 10 of which constitute the catalytic phosphatase domain. Detailed comparison of the position of intron/exon boundaries of the phosphatase domain of MPTP-PEST to those of several other protein tyrosine phosphatases indicates that the MPTP-PEST catalytic domain contains additional exons as a consequence of the insertion of novel introns. In addition, this analysis reveals a strong conservation of the genomic organization within the catalytic domain of the protein tyrosine phosphatase gene family. Finally, fluorescence in situ hybridization with MPTP-PEST genomic DNA refines the map position of MPTP-PEST to mouse chromosome 5A3 to B. This result is in agreement with the previous mapping of the human PEST gene to chromosome 7q11.23, a region of synteny with the centromeric portion of mouse chromosome 5. PMID- 7490088 TI - Cloning, structure, and chromosome localization of the mouse glutaryl-CoA dehydrogenase gene. AB - Glutaryl-CoA dehydrogenase (GCDH) is a nuclear-encoded, mitochondrial matrix enzyme. In humans, deficiency of GCDH leads to glutaric acidemia type I, an inherited disorder of amino acid metabolism characterized by a progressive neurodegenerative disease. In this report we describe the cloning and structure of the mouse GCDH (Gcdh) gene and cDNA and its chromosomal localization. The mouse Gcdh cDNA is 1.75 kb long and contains an open reading frame of 438 amino acids. The amino acid sequences of mouse, human, and pig GCDH are highly conserved. The mouse Gcdh gene contains 11 exons and spans 7 kb of genomic DNA. Gcdh was mapped by backcross analysis to mouse chromosome 8 within a region that is homologous to a region of human chromosome 19, where the human gene was previously mapped. PMID- 7490089 TI - High-resolution genetic and physical mapping of multiple epiphyseal dysplasia and pseudoachondroplasia mutations at chromosome 19p13.1-p12. AB - Multiple epiphyseal dysplasia (MED) and pseudoachondroplasia (PSACH) are autosomal dominant chondrodysplasias that have similar phenotypes at both clinical and cytological levels. With the recent mapping of PSACH and one form of MED (EDM1) to the pericentromeric region of chromosome 19, it is likely that the disease mutations are allelic. D19S212 and D19S215, genetic markers flanking the EDM1/PSACH locus, have been localized in a chromosome 19 physical map consisting of cosmid contigs ordered by high-resolution FISH. These two markers define an interval of approximately 3.1 Mb at the 19p13.1-p12 boundary. With as many as five informative crossovers within the D19S212-D19S215 interval in one family with EDM1 and one family with a mild form of PSACH, recombination mapping at greater resolution was undertaken. From cosmid contigs physically mapped within the D19S212-D19S215 interval, four new dinucleotide repeat polymorphisms have been identified. Analysis of recombinant haplotypes in the two families has narrowed the possible location of the EDM1/PSACH gene to an interval of approximately 600 kb. PMID- 7490090 TI - Developmental expression of p107 mRNA and evidence for alternative splicing of the p107 (RBL1) gene product. AB - Expression of p107, a protein with homology to the retinoblastoma tumor suppressor (pRB), was monitored during murine development. Northern blot tissue surveys identified two transcripts of 4.9 and 2.4 kb that hybridized to a p107 cDNA clone. Expression of both transcripts was detected in fetal tissues, with particularly high levels in the liver and heart. In contrast, p107 transcripts were markedly decreased in most adult tissues examined. Molecular cloning analyses revealed that the 4.9- and 2.4-kb transcripts encoded proteins with deduced molecular masses of 119 and 68 kDa, respectively. Genetic mapping studies suggested that the two p107 transcripts arose by alternative splicing of a common precursor. The protein encoded by the 2.4-kb transcript lacks the spacer and B motif of the "pocket domain," a region of homology between p107 and pRB that is required for binding to cell cycle regulatory proteins. Structural modifications resulting from alternative splicing may thus confer functional diversity upon the 119- and 68-kDa proteins. PMID- 7490091 TI - Generation of a transcription map at the HSD17B locus centromeric to BRCA1 at 17q21. AB - A detailed transcription map of the 320-kb region containing the HSD17B locus on chromosome 17 was generated. Thirty unique cDNA fragments, retrieved following the hybridization of immobilized YACs to primary pools of cDNAs prepared from RNA of mammary gland, ovary, placenta, and the Caco-2 cell line, were aligned into 10 transcription units by physical mapping and hybridization to RNAs of a series of tissues. The cDNAs were then further characterized by sequencing and used to screen mammary gland cDNA libraries. Fragments corresponding to the broadly expressed gamma-tubulin and Ki antigen genes were identified. A full-length cDNA clone encoding a 117-amino-acid protein homologous to the rat ribosomal protein L34 was isolated. Portions of genes with restricted patterns of expression were also obtained, including the previously characterized HSD17B1. One new gene, for which a full-length cDNA was isolated, was found to have an interesting tissue specific pattern of expression with abundant mRNA in both the colon and the testis and in the mammary carcinoma cell line BT-474. This contrasted with the barely detectable level observed in several tissues including normal mammary gland. Of the five additional transcription units identified, one showed no similarity, two showed identity to human expressed sequences, and two displayed similarity to genes of animal species by amino acid alignment. These latter cDNA clones include potential homologues of a rat nuclear tyrosine phosphatase and of a factor of Drosophila that is known to be involved in the negative regulation of transcription of segment identity genes. PMID- 7490092 TI - Identification of the human beta A2 crystallin gene (CRYBA2): localization of the gene on human chromosome 2 and of the homologous gene on mouse chromosome 1. AB - By using primers synthesized on the basis of the bovine beta A2 crystallin gene sequence, we amplified exons 5 and 6 of the human gene (CRYBA2). CRYBA2 was assigned to human chromosome 2 by concordance analysis in human x rodent somatic cell hybrids using the amplified PCR products as probe. Regional localization to 2q34-q36 was established by hybridizing the CRYBA2 probe to microcell and radiation hybrids containing defined fragments of chromosome 2 as the only human contribution. The CRYBA2 probe was also used to localize, by interspecific backcross mapping, the mouse gene (Cryba2) to the central portion of chromosome 1 in a region of known human chromosome 2 homology. Finally, we demonstrate that in both species the beta A2 crystallin gene is linked but separable from the gamma A crystallin gene. The beta A2 crystallin gene is a candidate gene for human and mouse hereditary cataract. PMID- 7490093 TI - Positional cloning of the nude locus: genetic, physical, and transcription maps of the region and mutations in the mouse and rat. AB - Mutations in the nude locus in mice and rats produce the pleiotropic phenotype of hairlessness and athymia, resulting in severely compromised immune system. To identify the causative gene, we utilized modern tools and techniques of positional cloning. Specifically, spanning the region in which the nude locus resides, we constructed a genetic map of polymorphic markers, a physical map of yeast artificial chromosomes and bacteriophage P1 clones, and a transcription map of genes obtained by direct cDNA selection and exon trapping. We identified seven novel transcripts with similarity to genes from Drosophila, Caenorhabditis elegans, rat, or human and three previously identified mouse genes. Based on our transcription mapping results, we present a novel approach to estimate the number of genes in a region and estimate that the nude locus resides in a region approximately threefold enriched for genes. We confirm a recently published report that the nude phenotype is caused by mutations in a gene encoding a novel winged helix or fork head domain transcription factor, whn (Nehls et al., Nature 372: 103-107, 1994). We report as well the mutations in the rat rnu allele and the complete coding sequence of the rat whn mRNA. PMID- 7490094 TI - Cloning, chromosomal localization, and physical linkage of the beta and gamma subunits (SCNN1B and SCNN1G) of the human epithelial amiloride-sensitive sodium channel. AB - Three subunits of the amiloride-sensitive Na+ channel, named alpha, beta, and gamma, have previously been cloned in rat colon. The human lung alpha chain (SCNN1A) has also been cloned and its gene localized on chromosome 12p13. We now report the molecular cloning of the human lung beta (SCNN1B) and gamma (SCNN1G) chains. In situ hybridization and pulsed-field electrophoresis experiments demonstrate that both genes are located within a common 400-kb fragment on chromosome 16p12-p13. Screening of the cDNA library reveals two forms of the beta subunit that differ by the presence or absence of a 464-bp fragment in the 3' region. A frameshift in the short form modifies the COOH terminal sequence of the corresponding protein. Since several similar frameshifts mutations have recently been reported in patients affected by a rare form of hypertension, the existence of COOH truncated forms of the beta chain might be of physiological importance. PMID- 7490097 TI - Mapping of the mouse homologue of the Wilson disease gene to mouse chromosome 8. AB - ATP7B, the gene altered in Wilson disease (WD) patients, lies in a block of homology shared between human chromosome 13q14 and the central region of mouse chromosome 14. However, we have mapped the murine homologue of ATP7B (Atp7b) to mouse chromosome 8 by somatic cell hybrid analysis. Analysis of 80 interspecific backcross offspring was used to position Atp7b close to D8Mit3 and another ATPase locus, Atp4b, on mouse chromosome 8. ATP4B lies in 13q34 and is separated from ATP7B by several loci whose mouse homologues map to mouse chromosome 14. The assignment of Atp7b to mouse chromosome 8 identifies a previously unrecognized region of homology between this chromosome and human chromosome 13. This assignment suggests a possible location for the toxic milk mutation in the mouse, which has been proposed as a homologue of WD. PMID- 7490095 TI - Genetic association between chromosome 8 microsatellite (MS8-134) and Werner syndrome (WRN): chromosome microdissection and homozygosity mapping. AB - Werner syndrome (WRN) is an autosomal recessive disorder characterized by premature aging that has been mapped to the short arm of chromosome 8, 8p11.2 p12. To refine the genetic map around the WRN region, we have isolated eight microsatellites for this region from a microdissection library. We typed members of Japanese families with WRN on the basis of homozygosity mapping analysis. There was no obligate recombination between the WRN locus and microsatellite clone, MS8-134 (D8S1055). The maximum lod score was 20.28 at theta = 0.00. Alleles for MS8-134 showed association with WRN in a case-control study (OR = 3.55, 95% CI 1.56-8.07, P < 0.01). Such microsatellites from a microdissection library of the definite chromosome region may be useful for positional cloning of the WRN gene. PMID- 7490096 TI - Chromosomal assignment of 20 cDNAs using flow-sorted spot-blot stamps. AB - Using a high-speed flow cytometer/sorter, we constructed spot-blot "stamps" measuring 3.5 x 2.0 cm containing 21 separate human chromosome fractions. Through hybridization to these stamps, 20 randomly selected cDNAs were assigned to specific chromosomes. Sequencing and BLAST database screening confirmed the location of one gene (UCHL1) and allowed the assignment of two other previously identified genes (LRP130 and cDNA IB871.) PMID- 7490099 TI - Mapping of the insulin promoter factor 1 gene (Ipf1) to distal mouse chromosome 5. AB - We have identified sequence variants in the 3' untranslated region of the insulin promoter factor 1 (Ipf1) cDNA sequence in mice and used a PCR-based oligonucleotide hybridization assay to map the Ipf1 gene to distal mouse chromosome (Chr) 5. An identical 12-bp insertion in the 3' untranslated region of the mouse Ipf1 gene sequence is present in Mus spretus and Mus m. musculus Czech II mice, two feral strains used for meiotic mapping by backcross analysis. The human IPF1 gene homolog would be expected to map to either chromosome 13q12-q14 or 7pter-q21 based on homology of synteny of other loci in this region of mouse Chr 5. Given its genetic map position, the Ipf1 gene may be part of a cassette of homeodomain-containing transcription factors involved in the development of the pancreas and other foregut-derived organs. PMID- 7490098 TI - Chromosomal localization, embryonic expression, and imprinting tests for Bmp7 on distal mouse chromosome 2. AB - Murine Bmp7 has been assigned to distal Chromosome 2 by interspecific backcross mapping. The map location suggests close linkage to classical mouse mutations and places Bmp7 within a chromosome region thought to contain one or more unidentified imprinted genes. A direct test suggests that Bmp7 is not imprinted. An examination of embryonic RNA expression patterns shows that Bmp7 is expressed in a variety of skeletal and nonskeletal tissues. Both embryonic expression patterns and the human chromosomal sublocalization inferred from its mouse location make Bmp7 a candidate for the gene affected in some patients with Holt Oram syndrome. PMID- 7490100 TI - Molecular cloning and chromosomal mapping of the mouse cyclin-dependent kinase 5 gene. AB - Cyclin-dependent kinase 5 (Cdk5) is predominantly expressed in neurons. In vitro, Cdk5 purified from the nervous tissue phosphorylates both high-molecular-weight neurofilament and microtubule-associated tau. The mouse gene encoding Cdk5 (Cdk5) was found to be 5 kb in length and divided into 12 exons. All of the exon-intron junctions matched the expected consensus sequence with the exception of the splice junction for intron 9, which has AT and AC dinucleotides instead of the usual GT and AG bordering sequence. In the 5'-flanking region of mouse Cdk5, several putative promoter elements were present, including AP1, Sp1, PuF, and TATA motifs. A metal regulatory element was also identified at position -207 to 201. Nucleotide sequence analysis of mouse Cdk5 showed high identity to the homologues of other vertebrate species, indicating that this kinase is highly conserved during evolution. Mouse Cdk5 was mapped to the centromeric region of mouse chromosome 5. PMID- 7490101 TI - Evidence for a novel exon in the coding region of the adenomatous polyposis coli (APC) gene. AB - Germline mutations of the tumor suppressor gene APC cause familial adenomatous polyposis. Somatic APC alterations are involved in several sporadic neoplasms, including colorectal, duodenal, gastric, and esophageal carcinoma. The APC mRNA is encoded by 15 exons. Additional transcripts have been reported, due to alternative splicing of coding as well as noncoding regions. Two mRNA isoforms occur due to a deletion of exon 7 or a partial deletion of exon 9. We have identified a novel exon, flanked by APC exons 10 and 11, which is expressed as an alternatively transcribed product of the gene. Further, we have shown that the novel exon consists of a heptad repeat motif and is conserved across species. PMID- 7490103 TI - The orphan nuclear receptor ROR alpha (RORA) maps to a conserved region of homology on human chromosome 15q21-q22 and mouse chromosome 9. AB - ROR alpha is a novel member of the steroid/thyroid/retinoid receptor superfamily with unique DNA-binding properties. We have mapped the RORA gene by fluorescence in situ hybridization to human chromosome 15q21-q22. To map the mouse Rora gene, a partial mouse cDNA clone was isolated from brain. Using interspecific backcross analysis, we have mapped the Rora gene to mouse chromosome 9. This places the human RORA gene in the proximity of the PML gene, which is involved in a reciprocal chromosomal translocation t(15:17) with the RARA gene in patients with acute promyelocytic leukemia. PMID- 7490102 TI - Genetic mapping of the mouse genes encoding the voltage-sensitive calcium channel subunits. AB - The genes encoding the alpha 1 and beta subunits of voltage-sensitive calcium channel were mapped in the mouse by analysis of the progeny of two multilocus crosses. The alpha 1, beta 2, and beta 4 subunit genes, termed Cchna1, Cchb2, and Cchb4, are located at different sites on proximal Chr 2, while the beta 3 subunit gene Cchb3 maps to Chr 15 near Wnt1. These results together with previous mapping data indicate that the calcium channel genes are dispersed in the mouse genome, unlike the sodium channel genes, which are clustered. PMID- 7490104 TI - Localization of the human phospholipid transfer protein gene to chromosome 20q12 q13.1. PMID- 7490105 TI - The human palmitoyl-CoA ligase (FACL2) gene maps to the chromosome 4q34-q35 region by fluorescence in situ hybridization (FISH) and somatic cell hybrid panels. PMID- 7490107 TI - Localization of the human OB gene (OBS) to chromosome 7q32 by fluorescence in situ hybridization. PMID- 7490106 TI - Mapping of a human LIM protein (CLP) to human chromosome 11p15.1 by fluorescence in situ hybridization. PMID- 7490108 TI - The gene coding for glial cell line derived neurotrophic factor (GDNF) maps to chromosome 5p12-p13.1. PMID- 7490109 TI - Assignment of the gastric inhibitory polypeptide receptor gene (GIPR) to chromosome bands 19q13.2-q13.3 by fluorescence in situ hybridization. PMID- 7490110 TI - The mouse Ftzf1 gene required for gonadal and adrenal development maps to mouse chromosome 2. PMID- 7490111 TI - Assignment of the human beta-adducin gene (ADD2) to 2p13-p14 by in situ hybridization. PMID- 7490112 TI - Signal transduction by B7/BB1 expressed on activated T lymphocytes: cross-linking of B7/BB1 induces protein tyrosine phosphorylation and synergizes with signalling through T-cell receptor/CD3. AB - We report here that B7/BB1 molecules expressed on activated T lymphocytes are involved in signal transduction. Anti-B7/BB1 monoclonal antibody (mAb) enhanced allogeneic proliferative responses against three different B lymphoma lines in a dose-dependent manner, while the same mAb inhibited T-cell response against allogeneic T cells expressing B7/BB1. Induction of B7/BB1 expression on T cells with allogeneic stimulation was confirmed by flow cytometric analysis. With the purified preactivated T cells as responder cells, anti-B7/BB1 mAb costimulated these primed T cells with coimmobilized anti-CD3 mAb. Moreover, cross-linking of B7/BB1 molecules induced protein tyrosine phosphorylation in preactivated T cells with a phosphorylation pattern distinct from those induced by signalling through other T-cell molecules. These results suggest that B7/BB1 molecules function not only as costimulatory ligands expressed on antigen-presenting cells but as receptors on T cells to transduce the costimulatory signals into the cells and may play a role for T-cell-T-cell interactions leading to clonal expansion of activated T lymphocytes. However, the physiological relevance of our finding remains to be explored. PMID- 7490113 TI - Oestrogen influences CD4+ T-lymphocyte activity in vivo and in vitro in beta 2 microglobulin-deficient mice. AB - Oestrogen directly influences autoimmune diseases and the immune response to microbes. We studied the effect of oestrogen on CD4+ T cells specific for lymphocytic choriomeningitis virus (LCMV) using mice genetically engineered to be deficient in beta 2-microglobulin (beta 2m-/-). These mice are deficient in beta 2-microglobulin, class I major histocompatibility complex (MHC) molecules and CD8+ T lymphocytes. Fatal leptomeningitis after intracranial infection with LCMV is mediated by CD8+ cytotoxic T lymphocytes (CTL) in wild-type C57BL/6 mice, and by CD4+ T cells in beta 2m-/- mice. Male and female wild-type C57BL/6 mice showed equal susceptibility to immune meningitis. In contrast, male beta 2m-/- mice were less susceptible to fatal immune meningitis than were females. Orchidectomy and oestrogen treatment of male beta 2m-/- mice in vivo restored susceptibility to meningitis. The classic weight loss seen in beta 2m-/- mice after intracranial infection was also accentuated in females. Further, the in vitro activity of CD4+ T cells from male beta 2m-/- mice, as measured by CTL assays, was shown to be dependent on oestrogen. The natural killer cell activity of spleen cells from beta 2m-/- mice after infection with LCMV was not affected by oestrogen. These data demonstrate the influence of oestrogen on CD4+ T-cell activity both in vivo and in vitro. PMID- 7490114 TI - Evidence for monoclonal expansion of synovial T cells bearing V alpha 2.1/V beta 5.5 gene segments and recognizing a synthetic peptide that shares homology with a number of putative autoantigens. AB - A peptide of 15 amino acids derived from the cereal glycine-rich cell wall protein (GRP), sharing a significant homology with Epstein-Barr virus nuclear antigen-1 (EBNA-1), fibrillar and procollagen, stimulated synovial fluid (SF) T cells from juvenile (JRA) and adult (RA) rheumatoid arthritis patients. An overexpression of the V alpha 2 gene family was found in the SF from patients who responded significantly to the peptide. To investigate in more detail the SF T cell responses to the GRP peptide, we established peptide-specific T-cell lines and clones from a DR8+ positive JRA patient with pauciarticular form. The T-cell clones were phenotyped as T-cell receptor (TCR)alpha beta+/CD4+ and their clonality was investigated by polymerase chain reaction (PCR) and flow cytometric analysis. TCR sequences from different clones demonstrated that the clones were identical and used the V alpha 2.1/J alpha 6 combined with V beta 5.5/J beta 2.7 gene segments. Interestingly, direct sequencing of the V alpha 2 family PCR product obtained from cDNA prepared from freshly isolated SF mononuclear cells identified the same TCR sequence as that used by the clones, suggesting the monoclonality of SF CD4+ T cells bearing V alpha 2.1/J alpha 6 gene products. The present data suggest a recruitment and expansion of a SF T-cell subpopulation, and also support the hypothesis that autoimmune diseases can be triggered by protein epitopes with crucial amino acids homologous to self-proteins. PMID- 7490115 TI - An immunogenetic analysis of the T-cell recognition of the major house dust mite allergen Der p 2: identification of high- and low-responder HLA-DQ alleles and localization of T-cell epitopes. AB - Cellular reactivity to Der p 2, a major allergen of the house dust mite (HDM) Dermatophagoides pteronyssinus, was studied in a group of 41 symptomatic HDM sensitive patients, using fresh peripheral blood mononuclear cells (PBMC) and assays of proliferation. Sixty per cent of the patients responded to Der p2, with reactivities being greater in patients with asthma as one of their clinical manifestations and also in those who had skin-test reactivity to a number of allergens. HLA-DR and -DQ serotyping was undertaken in 39 of the patients and the magnitude of T-cell proliferative responses to Der p 2 were found to be positively associated with DQ7 and negatively associated with DQ2. T-cell determinants within the Der p 2 molecule were identified by assays using a series of overlapping peptides (15- to 19-mers) spanning the entire protein. Fifty-nine per cent of the 41 HDM-sensitive patients responded to one or more of the peptides. All of the peptides were antigenic for at least one of the individuals, indicating the heterogeneity of the human repertoire reactive with Der p 2. There was a substantial variability in the number and location of epitopes recognized by T cells from the different allergic patients, the mean number per patient being 2.3 +/- 1.3 (SD). The most frequently recognized peptide was that spanning residues 111-129, being stimulatory in 66.7%, the other peptides were each recognized by between 8 to 25% of individuals. There was no correlation between the epitope recognized and the presence of particular HLA-DQ antigens. PMID- 7490116 TI - H-2-associated effects of flanking residues on the recognition of a permissive mycobacterial T-cell epitope. AB - Previously we have identified an immunodominant, eight-residue, epitope core sequence (TAAGNVNI) from the 19,000 MW protein of Mycobacterium tuberculosis, which is recognized in the context of multiple H-2 I-A molecules. In this study, the role of residues flanking this T-cell epitope core was examined, using a series of 20 mer analogue peptides in which the native flanking residues were progressively replaced with L-alanine. Analogue peptides were tested for their capacity to stimulate a CD4+ 19,000 MW protein-specific T-cell line, revealing that all but one N-terminal flanking residue could be replaced collectively by alanine without significant loss of stimulatory activity. However, clear H-2 associated differences in the requirement for flanking residues were demonstrated with peptide-specific T-cell hybridomas. In particular, H-2d-derived hybridomas were much more stringent in their requirement for flanking residues than were H 2b hybridomas. All polyalanine-substituted peptides bound I-Ab molecules, with affinities similar to the native unsubstituted peptide. In contrast, significantly reduced binding to I-Ad was observed with several analogue peptides, although without a clear relationship to the degree of substitution. Furthermore, in H-2b mice, neither immunogenicity nor cross-reactivity with the native peptide showed a clear inverse relationship with respect to the degree of alanine substitution. The results presented in this paper indicate that flanking residues can influence T-cell specificity and that these effects may be controlled by major histocompatibility complex (MHC) haplotype. PMID- 7490117 TI - The induction of renal autoantigen-specific T cells by a local Listeria monocytogenes infection. AB - In order to examine the possibility that a local chronic infection can induce organ-specific autoimmune disease, we inoculated unilateral kidneys with viable Listeria monocytogenes (intrarenal infection). The delayed footpad reaction against syngeneic kidney homogenate (KH) became positive from 1 week after initiating the intrarenal infection. A proliferative response of the spleen T cells from the infected mice was also observed against KH from 1 week after initiating the intrarenal infection, but no such response was seen against liver homogenate (LH). In contrast, an intravenous Listeria infection did not induce a delayed footpad reaction or proliferative response against KH, suggesting that these autoimmune responses were not caused by molecular mimicry between renal antigens and Listeria antigens. Furthermore, the ability to transfer the autoimmune response of spleen T cells from intrarenally infected mice was examined. The transferred mice showed a positive delayed footpad reaction against KH and an interstitial infiltration of mononuclear cells in their kidneys. These results demonstrate that the intrarenal Listeria infection induced renal autoantigen-specific T cells, which subsequently induced an autoimmune interstitial nephritis (AIN). The autoreactive T cells were all induced without immunization by autoantigens mixed with complete Freund's adjuvant. Based on these findings, we propose that a local bacterial infection may induce an autoimmune response against autoantigens in the infected organ and subsequently trigger organ-specific autoimmune disease. PMID- 7490118 TI - A protective role of gamma delta T cells in primary infection with Listeria monocytogenes in autoimmune non-obese diabetic mice. AB - We investigated the host defense mechanism in primary infection with Listeria monocytogenes in non-obese diabetic (NOD) mice at pre-diabetic stage showing an impaired responsiveness of the alpha beta T cells to T-cell receptor (TCR) triggering. The NOD mice showed a deteriorated resistance at the late stage after an intraperitoneal infection with L. monocytogenes compared with BALB/c and C57BL/6 mice as assessed by bacterial growth in organs. Consistent with our previous findings, a prominent increase in number of gamma delta T cells was evident at the early stage after infection, while generation of Listeria-specific alpha beta T cells was impaired in these mice. In vivo administration of anti-TCR gamma delta monoclonal antibody (mAb) allowed L. monocytogenes to grow exaggeratedly in the NOD mice. These results imply that gamma delta T cells may be mainly involved in protection against primary infection with L. monocytogenes in NOD mice. PMID- 7490119 TI - Conversion of Salmonella typhimurium to L-forms contributes to the maintenance of acquired immunity against murine typhoid. AB - Conversion of Salmonella typhimurium to L-forms, both in vitro and in vivo, resulted in the expression of proteins cross-reacting to the mycobacterial 65,000 MW heat-shock protein (hsp). Immunization of C3H/HeJ mice with a protective dose of stable L-form S. typhimurium induced gamma delta T cells in the liver, in accordance with the multiplication of L-form Salmonella in Kupffer cells. The number of gamma delta T cells decreased after the intracellular growth of L-form Salmonella plateaued. Persistance of the L-forms in Kupffer cells, however, allowed hepatic gamma delta T cells to increase within 48 hr of infection with virulent S. typhimurium. Thus, the intrahepatic colonization of L-form Salmonella seems to keep gamma delta T cells on standby, but the emergence of these T cells does not correlate with the expression of L-form hsp. In addition, Kupffer cells colonized by L-forms constitutively synthesized mRNA for interleukin-6 (IL-6) and tumour necrosis factor-alpha (TNF-alpha). These results suggest that conversion of S. typhimurium to L-forms in phagocytic cells builds up and maintains acquired resistance, conferred by live-cell vaccines of S. typhimurium, against murine typhoid. PMID- 7490120 TI - Regional variation in the proliferative rate and lifespan of alpha beta TCR+ and gamma delta TCR+ intraepithelial lymphocytes in the murine small intestine. AB - Using double staining for T-cell receptor (TCR) and 5-bromo-2'-deoxyuridine (BRdU) we have examined the proliferation rates and lifespan of murine intraepithelial lymphocytes (IEL's) in vivo. After a 24-hr pulse of BRdU the number of labelled alpha beta TCR+ IEL was significantly higher in the ileum than the duodenum. In contrast, incorporation of BRdU into gamma delta TCR+ IEL was significantly higher in the duodenum than the ileum. This regional variation was also seen after a 4-hr pulse of BRdU indicating that the differences probably reflect local rates of proliferation in the epithelium. Over a 6-day labelling period, the accumulation of labelled alpha beta TCR+ and gamma delta TCR+ IEL was linear, which allowed IEL lifespan to be calculated. There was considerable variation between groups of mice but the 50% population renewal time for alpha beta TCR+ IEL was 12-36 days in the duodenum and 9-11 days in the ileum, and for gamma delta TCR+ IEL was 12-21 days in the duodenum and 26-100 days in the ileum. The incorporation of BRdU into V beta 8+ IEL showed the same regional variation as alpha beta TCR+ IEL and the V delta 4 population behaved like the total gamma delta TCR+ IEL population. In contrast V beta 11+, potentially self-reactive IEL, showed a regional pattern of labelling like gamma delta TCR+ IEL. Incorporation of BRdU into both alpha beta TCR+ and gamma delta TCR+ IEL in germ-free mice was very low and did not show marked regional variation. alpha beta TCR+ and gamma delta TCR+ IEL from both proximal and distal bowel were cytotoxic. Therefore alpha beta TCR+ and gamma delta TCR+ IEL show different rates of division in different sections of the gut, perhaps reflecting responses to different antigens. Both alpha beta TCR+ and gamma delta TCR+ IEL reside in the epithelium for weeks during which time the gut epithelial population will have been renewed many times. PMID- 7490121 TI - Peptides of a major histocompatibility complex class I (Kb) molecule cause prolongation of skin graft survival and induce specific down-regulatory T cells demonstrable in the mixed lymphocyte reaction. AB - Six individual peptides of the major histocompatibility complex (MHC) class I molecule H-2Kb were synthesized. Intravenous injection of peptide 6 into mice prolonged the survival of Kb (BL/6 or B10.MBR) skin grafts on allogeneic R101 and B10.AKM mice, respectively. This was specific, as control skin grafts from Kk (B10.BR) or Kd (DBA/2) donors, respectively, were rejected at the same time in both control and peptide-treated mice. The optimal doses for peptide 6, which is from the alpha 2 domain, were defined. The test system was the inhibition of proliferation in vitro of naive lymph node cells by syngeneic mitomycin c-treated spleen cells from R101 mice preimmunized with irradiated stimulator splenocytes of Kb (BL/6) origin. Down-regulation was specific, as proliferation in response to third-party allogeneic stimulator Kk (B10.BR) splenocytes was not inhibited. Of the six peptides of H-2Kb tested, potent down-regulatory cells were induced by peptides 2 (alpha 1 domain) and 5 and 6 (alpha 2 domain). The greatest down regulatory activity was obtained by giving peptide 2 to mice that had already been immunized against H-2Kb by injecting EL4 cells. Under the same conditions, injecting peptide 2 did not induce any cytotoxic T cells. In contrast, specific cytotoxic lymphocytes (CTL) were induced when cells from primed mice were incubated for 4 days with heated stimulator cells from BL/6 mice. The data suggest that peptides from MHC class I molecules activate precursors of down regulatory T cells, but not of CTL, and this may explain their ability to prolong skin allograft survival. PMID- 7490122 TI - Enhancement of specific immunoglobulin production in SCID-hu-PBL mice after in vitro priming of human B cells with superantigen. AB - Priming of human mature B cells in vitro with staphylococcal enterotoxin A (SEA) prior to transplantation of the B cells into severe combined immunodeficiency (SCID) mice, together with human T-helper cells, resulted in higher and more uniform concentrations of serum IgG in the mice. This indicated that a large number of B cells had become activated, which was supported by the finding that SEA priming resulted in production of immunoglobulin displaying a more normal kappa/lambda ratio than was obtained in the absence of SEA priming. However, IgM concentrations were not affected by SEA priming. Immunization of mice, transplanted with SEA-primed B cells, with both primary and secondary antigens resulted in a high specific IgG response to both types of antigen. The elevated levels of specific antibodies were not merely the consequence of an unspecific stimulation of B cells caused by SEA, as the ratio of specific antibody to total IgG was much higher in animals receiving SEA-primed B cells. Thus, a co-operative effect on immunoglobulin production of stimulating B cells via surface immunoglobulin and help delivered by SEA-activated T-helper cells was indicated. A specific antigen-dependent IgM response to a secondary antigen was observed as well, but was, in contrast to the IgG response, not influenced by SEA priming of B cells. No IgM antibodies with reactivities to the primary antigens were detected in the SCID sera at any time-point after immunization. The results thus indicate that SEA might replace T-cell epitopes in antigens and efficiently recruit an abundance of T-cell help to B cells, resulting in enhanced production of specific IgG antibodies. PMID- 7490123 TI - Cytokine modulation of the immunosuppressive phenotype of pulmonary alveolar macrophage populations. AB - Under steady-state conditions, T-cell activation in the lung is tightly controlled by lymphocytostatic signals from resident pulmonary alveolar macrophages (PAM). The present study focuses upon the mechanism of suppression in the mouse, and how it is bypassed during local inflammatory challenge. Reactive nitrogen intermediates such as nitric oxide (NO) are shown to play a central role in the process as the expression of lymphocytostatic activity by resident murine PAM was abrogated by the NO synthetase inhibitor N-monomethyl-arginine. Overnight pretreatment of resident PAM with granulocyte-macrophage colony-stimulating factor (GM-CSF) abrogated lymphocytostatic activity, with a concomitant small decrease in NO production; this effect was markedly amplified by tumour necrosis factor-alpha (TNF-alpha), but the latter was ineffective alone. The cytokines were inactive if added singly or in combination to fresh PAM:T-cell co-cultures. If GM-CSF plus TNF-alpha exposure of PAM was prolonged beyond 48 hr, both lymphocytostatic and NO-producing capacity were spontaneously re-established. Transforming growth factor-beta (TGF-beta) also inhibited both NO production and lymphocytostatic activity of PAM, but in contrast to GM-CSF and TNF-alpha, TGF beta was only active if present throughout the PAM:T-cell coculture period. Additionally, monocytes recruited into the lung by a sterile inflammatory stimulus are shown to be initially stimulatory towards T-cell activation, and to progressively develop both T-cell suppressive- and NO synthetic-capacity as they mature into mature PAM in vivo. Thus, during acute lung inflammation, a series of overlapping mechanisms are potentially available to bypass local immunosuppression: secretion of cytokines which are capable of temporarily abrogating the immunosuppressive activity of resident PAM, and the recruitment of permissive monocytes which exhibit potent accessory cell activity, the net result being the creation of a transient 'window' for induction of local T cell-mediated immunity. PMID- 7490124 TI - Prevention of anti-T-cell receptor alpha beta monoclonal antibody-induced side effects by treatment with cyclosporin A without interference of monoclonal antibody-induced immunosuppression in mice. AB - Anti-T-cell receptor (TCR)alpha beta monoclonal antibody (mAb; H57-597) injection in mice caused cytokine (tumour necrosis factor and interferon-gamma) release and clinical side-effects such as piloerection and body weight loss similar to anti CD3 mAb (145-2C11) injection. Treatment with cyclosporin A (CsA) for 3 days, from day -2 to day 0, prior to anti-TCR alpha beta mAb injection almost completely abolished the mAb-induced cytokine release, and completely inhibited the mAb induced body weight loss. Furthermore, treatment with CsA from day -2 to day 0 did not inhibit the mAb-induced in vivo immunosuppressive effects, i.e. prolongation of skin allograft and T-cell depletion in the periphery. These results indicate that CsA treatment prior to mAb treatment could effectively inhibit the mAb-induced side-effects without interference of the mAb-induced in vivo immunosuppression. From these results, we propose that CsA treatment prior to injection of anti-TCR alpha beta mAb may be recommended to reduce mAb-induced side-effects. PMID- 7490126 TI - Expression of interferon-gamma receptors on murine oligodendrocytes and its regulation by cytokines and mitogens. AB - Interferon-gamma (IFN-gamma) is a cytokine known to exert an important immunological role on astrocytes and oligodendrocytes. As a receptor for IFN gamma has been demonstrated on murine astrocytes, we have searched for a specific receptor on the cell surface of pure mouse oligodendrocytes maintained in tissue culture. Using recombinant murine IFN-gamma labelled with 125I, we have established the basic physicochemical parameters of the binding. A single receptor was found with a Kd of 1 x 10(-9) M. The number of receptors per cell was 3000-4000 and its molecular weight, as determined by cross-linking experiments, is 87,000. The binding of IFN-gamma to its oligodendrocyte receptor is saturable, specific and temperature-dependent. The receptor-IFN-gamma complex is quickly endocytosed at 37 degrees (the half-time of maximal internalization is around 1 min). Some cytokines, such as interleukin-1 alpha and interleukin-6, up regulated the expression of the oligodendrocyte receptor, but others, such as tumour necrosis factor-alpha, did not. A dramatic increase in receptor expression is induced by lipopolysaccharide but it is not detectable after treatment with concanavalin A. PMID- 7490125 TI - Induced expression of mRNA for IL-5, IL-6, TNF-alpha, MIP-2 and IFN-gamma in immunologically activated rat peritoneal mast cells: inhibition by dexamethasone and cyclosporin A. AB - We examined the capacity of purified rat peritoneal connective tissue-type mast cells (PMC) to express mRNA for several cytokines. Stimulation of PMC with anti IgE for 4 hr induced the expression of mRNA encoding interleukin-5 (IL-5), IL-6, tumour necrosis factor-alpha (TNF-alpha), macrophage inflammatory protein-2 (MIP 2) and interferon-gamma (IFN-gamma). Unstimulated PMC expressed detectable mRNA for TNF-alpha but not for the other four cytokines. Incubation of PMC with cyclosporin A (CsA) or dexamethasone (DEX), each at 10(-6) M for 24 hr, significantly inhibited the induced expression of mRNA for each of the five cytokines, and also inhibited release of biologically active TNF-alpha. Throughout these experiments mRNA levels of the housekeeping gene G3PDH were not altered by stimulation with anti-IgE or incubation with CsA or DEX. We conclude that immunological activation of rat PMC induces gene expression of several cytokines and that expression of these genes can be inhibited by immunosuppressive drugs. PMID- 7490127 TI - Tumour necrosis factor, but not interferon-gamma, is essential for acquired resistance to Listeria monocytogenes during a secondary infection in mice. AB - Mice with a secondary Listeria monocytogenes infection eliminate the bacteria much faster and more efficiently from their organs than mice with a primary infection. During the course of a secondary infection, serum concentrations of interferon-gamma (IFN-gamma) and tumour necrosis factor-alpha (TNF) are higher than during a primary infection. The aim of the present study was to determine whether these cytokines are involved in the acquired resistance to L. monocytogenes during a secondary infection in mice. In order to neutralize cytokines, alginate-encapsulated cells, which form anti-cytokine monoclonal antibodies, were injected into the nuchal region of mice during a Listeria infection. Mice recovered from a sublethal primary Listeria infection, which acquired cell-mediated immunity, received a subcutaneous injection of anti-IFN gamma-forming cells, or anti-TNF-forming cells, and 4 days later received an intravenous injection with 10 50% lethal dose (LD50) L. monocytogenes. The number of bacteria recovered from the liver and spleen of immune mice treated with anti IFN-gamma-forming cells was slightly larger (approximately 1 log10) than that found for immune mice treated with anti-beta-galactosidase-forming cells, called immune control mice. The organs of immune mice treated with anti-TNF-forming cells yielded significantly more (approximately 4 log10) bacteria than those of immune control mice, more than those of immune mice treated with anti-IFN-gamma forming cells, and comparable numbers to those of non-immune mice. Taken together, these results demonstrate that TNF is essential in acquired resistance to L. monocytogenes during a secondary infection in mice, while IFN-gamma plays a minor role. PMID- 7490128 TI - Transforming growth factor-beta down-regulates major histocompatibility complex class I antigen expression and increases the susceptibility of uveal melanoma cells to natural killer cell-mediated cytolysis. AB - Intraocular melanomas, especially those of the anterior segment, reside within an immunologically privileged milieu. Aqueous humour contains a variety of immunomodulatory factors that are believed to contribute to ocular immune privilege. Among these is transforming growth factor-beta (TGF-beta), which has been shown to down-regulate major histocompatibility complex (MHC) class I antigens on normal cells. Since the susceptibility of tumour cells to natural killer (NK) cell-mediated lysis is inversely correlated with the expression of MHC class I antigens, tumour cells exposed to TGF-beta might be expected to experience enhanced susceptibility to NK-mediated killing. This was examined by incubating two human uveal melanoma cell lines in the presence of TGF-beta and evaluating the expression of MHC class I antigen and susceptibility to NK cell mediated lysis. OCM1 and OCM8 melanoma cells constitutively express high levels of class I antigen (85-90% positive) and low susceptibility to NK-mediated lysis in vitro (3-8%). Incubation with TGF-beta produced a significant reduction in class I antigen expression (52-62%) and a proportional increased susceptibility to NK cell-mediated cytolysis (17%). Analogous effects were found using a human uveal melanoma cell line (OCM3) that constitutively expresses low amounts of class I (< 5% positive) and high NK susceptibility (35% lysis). Stimulation of class I antigen expression by incubation with interferon-gamma resulted in a sharp increase in class I expression (80% positive) and a comparable diminution in susceptibility to NK cell-mediated lysis (< 10%). The results indicate that TGF-beta, at concentrations found in the aqueous humour, can significantly alter MHC class I antigen expression and the susceptibility of ocular melanoma cells to NK cell-mediated cytolysis. PMID- 7490129 TI - Extracellular matrix proteins and integrin receptors in reactive and non-reactive lymph nodes. AB - The extracellular matrix (ECM) proteins collagen I, III and IV, laminin, fibronectin, vitronectin, thrombospondin, tenascin and their integrin receptors of the beta 1 and beta 3 subfamilies showed characteristic patterns of distribution in different compartments of non-reactive and reactive lymph nodes (human and monkey). This was particularly evident during development of germinal centres. Thus, ECM proteins (collagens, laminin, fibronectin and tenascin) were abundant in the interfollicular (T-cell rich) compartments of non-reactive as well as reactive lymph nodes. In primary follicles, collagen I, III and fibronectin were expressed but displaced by the expanding germinal centre during the formation of secondary follicles in reactive lymphoid tissues. The integrin subunits were mainly associated with endothelial cells and lymphoid cells in interfollicular areas, but were absent or only poorly expressed in primary as well as secondary follicles. Evidently the expression of ECM components and their integrin receptors is markedly down-regulated in the reactive, highly proliferative germinal centres. PMID- 7490130 TI - Regulation and function of adhesion molecule expression by human alveolar epithelial cells. AB - The role of major histocompatibility complex (MHC) and adhesion molecule expression by alveolar epithelium on the modulation of immune responses in the lung is not understood. We have developed efficient methods to isolate, purify and culture human alveolar epithelial cells (type II pneumocytes) in vitro. The expression of MHC and adhesion molecules by isolated, cultured and cytokine stimulated alveolar epithelial cells was quantified by flow cytometry, and demonstrated the presence of T-cell ligands including class I MHC, HLA-DR and HLA DP, intracellular adhesion molecule-1 (ICAM-1; CD54) and lymphocyte function associated antigen (LFA-3; CD58), but not vascular cell adhesion molecule-1 (VCAM 1) (CD106) or B7 (CD80). The proinflammatory cytokine interferon-gamma (IFN gamma) caused an up-regulation of class I MHC and ICAM-1. In contrast, tumour necrosis factor-alpha (TNF-alpha) and interleukin-1 beta (IL-1 beta) had little effect on the expression of these surface antigens by human alveolar epithelial cells. The functional activity of alveolar epithelial adhesion molecules was then studied by determining their ability to bind allogeneic lymphocytes. An increase in lymphocyte adherence to monolayers of alveolar epithelial cells was observed following in vitro activation. However, up-regulation of alveolar epithelial counter receptors with the proinflammatory cytokine gamma-IFN did not enhance adhesion. The adhesive interaction between CD18 on lymphocytes and ICAM-1 on alveolar epithelial cells was demonstrated by the use of blocking antibodies specific for both ligands. Blockade of LFA-3 on alveolar monolayers also suppressed lymphocyte adherence. In conclusion, alveolar epithelial cells expressed MHC HLA-A, B, C, HLA-DR and -DP, and functional adhesion molecules including ICAM-1 and LFA-3. PMID- 7490131 TI - Sensitization of MHC class I-restricted T cells to exogenous proteins: evidence for an alternative class I-restricted antigen presentation pathway. AB - Immunization with exogenous proteins usually fails to immunize CD8+ T cells in vivo. Here we report that chicken ovalbumin (OVA) denatured by heat or sodium dodecyl sulphate (SDS) effectively induced CD8+ cytolytic T cells in vivo. The cytolytic T-lymphocyte (CTL) population generated recognized syngeneic target cells pulsed with the immunodominant OVA peptide (257-264) or transfected with the OVA protein-encoding gene. To analyse the mechanisms of how denatured OVA enters the class I-restricted pathway of antigen presentation, we took advantage of the fact that denatured OVA sensitizes target cells in vitro for lysis by OVA specific CTL. We found that neither inhibition of protein synthesis (by cycloheximide) nor blocking of transport via the Golgi apparatus (by brefeldin A) interfered with the class I-restricted presentation of denatured OVA in vitro. In addition, transporter associated with antigen presentation (TAP)-dependent transport into the endoplasmic reticulum (ER) was not required for effective presentation, as TAP-deficient cells (RMA-S) could be sensitized effectively by denatured OVA for recognition by class I-restricted CTL. In contrast, class I restricted presentation of denatured OVA was sensitive to lysosomotropic agents (NH4Cl, vinblastine and leupeptin), indicating that endosomal-like compartments are involved in the presentation of denatured OVA. Sensitization was inhibited at low temperature, yet took place in the presence of sucrose and in the absence of K+, indicating that denatured OVA enters the cell via fluid-phase endocytosis. Hence the results provide further evidence for an alternative class I-restricted pathway of antigen presentation for exogenous proteins. As that pathway seems to be effective in vivo, it offers a new and effective way of vaccination of CD8+ CTL. PMID- 7490132 TI - Migration of human antigen-presenting cells in a human skin graft onto nude mice model after contact sensitization. AB - Fluorescent contact chemical allergens provoke sensitization after application on both syngeneic and allogeneic skin grafts in mice. We attempted to determine whether the functional activity in a contact sensitization response of human skin graft was affected at the level of antigen uptake and migration. After xenogeneic skin transplantation, we examined the effect of topical exposure of the graft to rhodamine B isothiocyanate (RITC). This paper describes the migration of RITC carrying cells and human major histocompatibility complex (MHC) class II DR (HLA DR)+ cells, from the graft to mouse draining lymph nodes. As demonstrated by immunohistochemistry, grafting resulted in a time-dependent decrease of human HLA DR+ and CD1a+ cells, and an increase of mouse MHC class II (Ia)+ cells within the graft. Application of RITC on a 3-week-old human skin graft showed optimal migration capability compared to 6- or 9-week-old grafts. In addition, the time dependent increase of frequencies of RITC+ and HLA-DR+ cells in the draining lymph nodes, and the time-dependent decrease of HLA-DR+ cells in the 3-week-old human skin graft, were concurrent. Supporting these data, human cytokine interleukin-1 alpha (IL-1 alpha), IL-1 beta and tumour necrosis factor-alpha (TNF alpha), analysis in situ revealed that cytokine production by keratinocytes, a property associated with dendritic cell migration, was preserved in the human skin graft. Thus, like dendritic cells in contact sensitization in allografted skin, dendritic cells from human xenografted skin onto nude mice are capable of migration to mouse draining lymph nodes after allergen application. Induction of contact hypersensitivity is possible in a human skin graft onto nude mice model, although the use of this ex vivo model to analyze contact sensitivity is probably limited to 3 weeks after transplantation. PMID- 7490133 TI - Inhibition of activation of human T lymphocytes by the complement C1 esterase inhibitor. AB - The major histocompatibility complex (MHC) class I molecules have been shown to be substrates to both of the complement C1 esterases. The effect of a C1 esterase mediated cleavage of the MHC class I molecules on the activation process of lymphocytes was investigated by including the complement C1 esterase inhibitor (C1-Inh) in the medium during activation of human peripheral lymphocytes by staphylococcal enterotoxin A (SEA). The C1-Inh was shown to inhibit the activation of both CD4+ and CD8+ cells. No effect on activation of B lymphocytes was recorded, although the complement C1 complex was shown to bind to the B lymphocytes. Furthermore, the C1 complex bound to mononuclear cells was shown to be cleaved into molecular weights corresponding to the activated forms of the C1 esterases. The effect of the C1-Inh was much more pronounced at low cell density and the inhibition was not affected by the addition of interleukin-2 (IL-2). However, the inhibition was reduced when the cells were disturbed by addition of new portions of C1-Inh, 24 and 48 hr after the initiation of the activation. This indicates that the C1-Inh interference with the activation of T lymphocytes is mediated through a mechanism that requires some form of cell contact. PMID- 7490134 TI - Characterization of a glycosyl-phosphatidylinositol anchor-deficient subline of Raji cells. An analysis of the functional importance of complement inhibitors on the Raji cell line. AB - Analysis of complement inhibitory proteins present on the surface of Raji cells (obtained from the European Collection of Animal Cell Cultures; originally established from human Burkitt's lymphoma) revealed two populations of cells. These populations differed in their expression of the glycosyl phosphatidylinositol (GPI)-anchored inhibitors CD59 and decay-accelerating factor (DAF). Two stable clones were established by limiting dilution of the original cell culture. Raji+3 expressed CD59 and DAF whereas Raji-26 expressed neither inhibitor. Both clones expressed membrane cofactor protein (MCP). Analyses of other cell surface proteins (CD19, CD35, CD48 and CD58 (transmembrane form)) revealed similar levels of expression of transmembrane proteins by both clones. However, CD48 was expressed only by Raji+3. As CD48, DAF and CD59 are all GPI anchored molecules it is likely that a defect in the GPI-anchoring mechanism is responsible for the generation of the second population of cells. The two clones demonstrated markedly different sensitivities to complement. When equally sensitized cells from both clones were treated with normal human serum (12.5%) for 1 hr at 37 degrees, the Raji+3 clone was resistant to complement-mediated lysis, whereas approximately 70% of the Raji-26 cells were lysed. However, by using specific antibody to block the function of membrane-bound complement inhibitors, lysis of Raji+3 was demonstrated. Whilst blocking of one inhibitor only on the cell had little effect on cell killing, blocking of two or more inhibitors significantly increased cell lysis. Our results demonstrated that all three inhibitors expressed by these cells contributed to protection against classical pathway-mediated complement activation. However, whilst a limited protective role was seen for MCP, CD59 and DAF appeared to be of far more importance for protection from complement-mediated lysis via the classical pathway. PMID- 7490135 TI - The N-terminal end of the CH2 domain of chimeric human IgG1 anti-HLA-DR is necessary for C1q, Fc gamma RI and Fc gamma RIII binding. AB - We have found that amino acid residues necessary for C1q and Fc gamma R binding of human IgG1 are located in the N-terminal region of the CH2 domain, residues 231-238, using a matched set of engineered antibodies based on the anti-HLA-DR antibody L243. Changing the leucine 235 in the CH2 region of IgG3 and IgG4 to glutamic acid was already known to abolish Fc gamma RI binding. We have confirmed this for IgG1 and also found a concomitant abolition of human complement lysis with retention of Fc gamma RIII-mediated function. Changing the glycine at 237 to alanine of IgG1 also abolished Fc gamma RI binding and reduced human complement lysis and Fc gamma RIII-mediated function. Exchanging the whole region 233-236 with the sequence found in human IgG2, abolished Fc gamma RI binding and human complement lysis and reduced Fc gamma RIII-mediated function of IgG1. In contrast, a change in the previously described C1q-binding motif, from lysine at 320 to alanine, had no effect on IgG1-mediated complement lysis. PMID- 7490136 TI - Gene expression of angiotensin II receptor in blood cells of Cushing's syndrome. AB - The relation between serum cortisol, plasma renin activity, angiotensin II (Ang II), or aldosterone levels and peripheral blood cell (mononuclear leukocytes and platelets) angiotensin II type 1A (AT1A) and 1B (AT1B) receptor mRNA levels was examined in both patients with Cushing's syndrome (seven patients with Cushing's syndrome due to unilateral adrenal cortical adenoma) and control subjects (seven normotensive patients with renal cell carcinoma). Blood was collected from each participant for estimation of plasma renin activity and plasma angiotensin II, aldosterone, and cortisol concentrations and for isolation of mononuclear leukocytes and platelets, which were then used to measure AT1A and AT1B receptor mRNA levels before and after adrenalectomy with the use of reverse transcription polymerase chain reaction. In patients with Cushing's syndrome, both mononuclear leukocyte and platelet AT1A mRNA levels, which were elevated, were reduced after removal of the adrenal tumors, whereas AT1B receptor mRNA levels of both types of blood cells did not significantly change after adrenalectomy. In contrast, in control subjects, both AT1A and AT1B receptor mRNA levels did not significantly change after unilateral adrenalectomy and nephrectomy. In the adrenal tumors of patients with Cushing's syndrome, gene expression of AT1A receptor was decreased compared with that from adrenals of control subjects. AT1A receptors of the platelets were shown to be upregulated in a manner similar to those of mononuclear leukocytes in patients with Cushing's syndrome. These results suggest that cortisol excess is an important factor upregulating AT1A receptor mRNA levels in human blood cells. PMID- 7490137 TI - Renal artery stenosis rapidly enhances atrial natriuretic peptide gene expression. AB - The aim of this study was to examine the influence of the systemic renin angiotensin system on the gene expression of atrial natriuretic peptide in rat hearts. The renin-angiotensin system was stimulated (1) by unilateral renal artery clipping (0.2-mm clip, 2 days), producing a fourfold increase of circulating plasma renin activity and increasing blood pressure; (2) by furosemide infusion with simultaneous salt substitution, increasing plasma renin activity values to 45 ng angiotensin I/h per milliliter without changing blood pressure; or (3) by administration of the calcium antagonist amlodipine, which increased plasma renin activity values to 42 ng angiotensin I/h per milliliter and lowered blood pressure. Unilateral renal artery clipping increased atrial natriuretic peptide mRNA levels approximately 20-fold in the left ventricles and approximately twofold in the right ventricles and atria. Furosemide infusion had no effect on cardiac atrial natriuretic peptide mRNA levels, and in amlodipine treated rats, cardiac atrial natriuretic peptide mRNA levels decreased to 30% of control values. The increase of atrial natriuretic peptide mRNA in the ventricles during renal artery clipping was blunted by the administration of the angiotensin converting enzyme inhibitor ramipril, which also attenuated the blood pressure rise. In clipped rats amlodipine did not change elevated plasma renin activity values but abolished the rise of blood pressure and also attenuated the rise of atrial natriuretic peptide mRNA in the hearts. These findings indicate that an increase of the activity of the systemic renin-angiotensin system does not result in an obligatory change in cardiac atrial natriuretic peptide gene expression.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7490138 TI - Human atrial natriuretic peptide gene delivery reduces blood pressure in hypertensive rats. AB - Chronic infusion of atrial natriuretic peptide (ANP) has been shown to cause natriuresis, diuresis, and hypotension in rats and humans. We explored the effect of a continuous supply of ANP by somatic ANP delivery on genetically hypertensive rats. A DNA construct containing the human ANP gene fused to the Rous sarcoma virus 3'-long terminal repeat (RSV-LTR) was injected intravenously into spontaneously hypertensive rats (SHR) through the tail vein. Expression of human ANP in SHR was identified in the heart, lung, and kidney by radioimmunoassay and reverse transcription-polymerase chain reaction followed by Southern blot analysis. A single injection of naked ANP plasmid DNA (12.3 kb) caused a significant reduction of systemic blood pressure in young SHR (4 weeks old), and the effect continued for 7 weeks. The differences were significant at 1 to 2 weeks (n = 6, P < .05) and 3 to 6 weeks after injection (n = 6, P < .01) A maximal blood pressure reduction of 21 mm Hg in young SHR was observed 5 weeks after injection with ANP DNA (159.4 +/- 3.02 mm Hg, mean +/- SEM, n = 6) compared with SHR injected with vector DNA alone (180.2 +/- 3.02 mm Hg, mean +/- SEM; n = 6; P < .01). Somatic gene delivery of human ANP DNA had no effect on the blood pressure of adult SHR (12 weeks old). After ANP gene delivery, there were significant increases in urinary volume and urinary potassium output (n = 6, P < .05) but not in body weight, heart rate, water intake, urinary sodium output, urinary creatine, and urinary protein.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7490139 TI - Role of superoxide in the depressed nitric oxide production by the endothelium of genetically hypertensive rats. AB - We undertook these studies to determine whether a deficient nitric oxide production in genetically hypertensive rats could result from its being scavenged by an excess production of superoxide. In one study we used a porphyrinic microsensor to measure nitric oxide concentrations released by cultured endothelial cells from stroke-prone spontaneously hypertensive rats (SHRSP) and normotensive Wistar-Kyoto rats (WKY). SHRSP cells released only about one third the concentration of nitric oxide as did WKY cells. Treatment of cells with superoxide dismutase increased nitric oxide release, demonstrating that normally nitric oxide is scavenged by endogenous superoxide. The increase in nitric oxide release in response to superoxide dismutase treatment was more than twice as great from SHRSP as from WKY cells, demonstrating the greater amount of superoxide in the hypertensive rats. A direct measure of superoxide with the use of lucigenin demonstrated the presence of 68.1 +/- 7.1 and 27.4 +/- 3.5 nmol/L of this anion in SHRSP and WKY endothelial cells, respectively. The presence of superoxide in the rat aorta was also estimated by quantification of its effect on carbachol relaxation. This relaxation was diminished when endogenous superoxide dismutase was blocked by diethyldithiocarbamic acid. This blockade reduced the relaxation by 51.2 +/- 5.2% in SHRSP aortas and by only 22.0 +/- 8.2% (P = .015) in WKY aortas. Data from these diverse systems are in agreement that superoxide production is excessive in SHRSP tissues. This excess superoxide, by scavenging endothelial nitric oxide, could contribute to the increased vascular smooth muscle contraction and hence to the elevated total peripheral resistance of these rats. PMID- 7490140 TI - Does keloid pathogenesis hold the key to understanding black/white differences in hypertension severity? AB - Repeated national surveys have shown that in the United States hypertension is more common and more severe in the black than in the white population. This discussion presents the hypothesis that the racial difference in hypertension severity is because of differences in growth factors (cytokines) affecting vascular smooth muscle cell growth. This hypothesis is derived from studies of keloids, which occur almost exclusively in blacks. Keloid fibroblasts in culture have growth characteristics that differ substantially from those of normal skin. Furthermore, there is evidence that peripheral blood mononuclear cells from patients with keloids produce different amounts of cytokines than do similar preparations from the blood of individuals without keloids. A growing body of evidence indicates that growth factors play a pivotal role in the pathogenesis of atherosclerosis, in experimental forms of hypertension, and in various renal diseases; it may be that they function in clinical hypertension as well. PMID- 7490141 TI - Effect of copper-zinc superoxide dismutase on endothelium-dependent vasodilation in patients with essential hypertension. AB - Patients with essential hypertension have abnormal endothelium-dependent vasodilation related to decreased nitric oxide activity. The specific mechanism responsible for this abnormality is unknown. Recent studies in hypertensive animals have suggested an augmented destruction of nitric oxide by superoxide anions. Therefore, in the present study we aimed to investigate whether this mechanism is responsible for the abnormal vasodilator function of hypertensive patients. To this end, we studied the vascular responses to acetylcholine (an endothelium-dependent vasodilator) and sodium nitroprusside (a direct smooth muscle dilator) before and after combined administration of copper-zinc superoxide dismutase (a scavenger of superoxide anions with poor intracellular penetrance; 6000 U/min) in 20 healthy control subjects (11 men and 9 women; aged 50 +/- 6 years) and 20 hypertensive patients (13 men and 7 women; aged 51 +/- 9 years). Drugs were infused into the brachial artery, and the response of the forearm vasculature was measured by plethysmography. The vasodilator response to acetylcholine was significantly blunted in hypertensive patients compared with control subjects (maximal flow: 8.2 +/- 4 versus 12.7 +/- 3 mL/min per 100 mL; P < .02); however, no difference was observed in the response to sodium nitroprusside (8.1 +/- 4 versus 9.5 +/- 3 mL/min per 100 mL). In healthy control subjects superoxide dismutase infusion did not modify the vasodilator response to acetylcholine (maximal flow: 12.7 +/- 3 before versus 12.1 +/- 3 after superoxide dismutase). Similarly, in hypertensive patients superoxide dismutase infusion did not alter the response to acetylcholine (maximal flow: 8.2 +/- 4 versus 7.7 +/- 4).(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7490143 TI - The Egyptian National Hypertension Project (NHP). Design and rationale. The NHP Investigative Team. AB - Hypertension and its complications appear to be increasingly common in Egypt. The National Hypertension Project (NHP) is a collaborative Egyptian-American effort with the following objectives: (1) to determine the prevalence of hypertension and blood pressure-related preclinical and clinical complications in Egyptian adults, (2) to identify environmental factors associated with high blood pressure, and (3) to build an infrastructure for research and education in cardiovascular disease prevention in Egypt. The NHP surveys were conducted in six of Egypt's 26 governorates, representing distinct geographic regions. In each of these six governorates, a probability sample of 600 households was surveyed. NHP was conducted in two phases. In phase I a team of specially trained physicians conducted household surveys in which all adult residents (age > or = 25 years) were screened for hypertension. In phase II hypertensive adults identified in phase I (ie, those with systolic pressure > or = 140 mm Hg and/or diastolic pressure > or = 90 mm Hg and/or those receiving antihypertensive drug therapy) along with a randomly selected sample of normotensive control subjects were asked to participate in a more extensive evaluation. This included a detailed history and clinical examination, an electrocardiogram, an echocardiogram, laboratory studies, and skin color reflectance (on a subsample). As the first systematic, national survey of hypertension and its complications in an Arab country, NHP should provide data of great interest to the scientific, provider, and public health communities. PMID- 7490142 TI - Diabetes mellitus and associated hypertension, vascular disease, and nephropathy. An update. AB - Because considerable important information has been published since our previous review, this update concentrates on new findings with regard to cardiovascular and renal risk factors contributing to the striking morbidity and mortality of these coexisting diseases. For example, a large body of investigative data has recently emerged suggesting or delineating a pathogenic role for hyperglycemic related glycosylation and oxidation of lipoproteins and vascular and renal tissues. Great strides have recently been made in the understanding of platelet, coagulation, lipoprotein, and endothelial abnormalities in the pathogenesis of cardiovascular and renal disease associated with diabetes mellitus and hypertension. Major progress has been made in clarifying the pathophysiology of glomerulosclerosis and other processes involved in the progression of diabetic nephropathy. Furthermore, accumulating data surveyed in this review address new and promising pharmacological interventions that specifically address these pathophysiological mechanisms. PMID- 7490144 TI - Hypertension prevalence, awareness, treatment, and control in Egypt. Results from the Egyptian National Hypertension Project (NHP). NHP Investigative Team. AB - This report from the Egyptian National Hypertension Project presents national estimates of the prevalence of hypertension and the extent to which high blood pressure is being detected, treated with medications, and controlled in the Egyptian population. The results are based on findings from a national probability survey of adults > or = 25 years of age conducted in six Egyptian governorates. With the use of a stratified multistage probability design, 6733 people (85% response rate) were examined. Hypertension was defined as systolic pressure > or = 140 mm Hg, and/or diastolic pressure > or = 90 mm Hg, and/or reported treatment with one or more antihypertensive medications. Overall, the estimated prevalence of hypertension in Egypt was 26.3%. Hypertension prevalence increased progressively with age, from 7.8% in 25- to 34-year-olds to 56.6% in those 75 years or older. Hypertension was slightly more common in women than in men (26.9% versus 25.7%, respectively). Overall, 37.5% of hypertensive individuals were aware that they had high blood pressure, 23.9% were being treated with antihypertensive medications, and 8.0% were under control (systolic pressure < 140 mm Hg and diastolic pressure < 90 mm Hg). Hypertension prevalence as well as awareness, treatment, and control rates varied by region, with Cairo having the highest prevalence (31.0%) and the Coastal Region having the highest control rate (15.9%). Rates of awareness, treatment, and control tended to be lowest in areas of lower socioeconomic status. Our results indicate that hypertension is highly prevalent in Egypt and that the rates of hypertension is awareness, treatment, and control are relatively low.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7490145 TI - Expression of rat kallikrein and epithelial polarity in transfected Madin-Darby canine kidney cells. AB - Many properties of urinary kallikrein are well characterized, but the intracellular processing of prokallikrein and release by kidney cells have yet to be clarified. We report here on the synthesis of prokallikrein in Madin-Darby canine kidney (MDCK) cells transfected with rat submaxillary gland kallikrein cDNA and on its activation by MDCK cells and by an enriched liver Golgi membrane preparation. Transfected MDCK cells secreted only prokallikrein at both the apical and basolateral sides in about a 4:1 ratio, but cells transfected with kallikrein cDNA in reverse orientation or untreated cells released only traces of the enzyme. Prokallikrein, in culture medium or in homogenized MDCK cells, was fully activated by trypsin but activated only to 44% by thermolysin. Prokallikrein was synthesized and released into the medium at a high rate: the enzyme secreted by 5 x 10(6) cells in 24 hours cleaved 46 nmol/min D-Val-Leu-Arg 7-amino-4-methylcoumarin and liberated 63 ng/min bradykinin after activation. Immunocytology indicated the association of prokallikrein with the Golgi apparatus in the transfected cells. Antiserum to rat urinary kallikrein detected a single band in a Western blot of conditioned medium and also immunoprecipitated the enzyme. Aprotinin inhibited activated prokallikrein. Although MDCK cells released prokallikrein, their homogenates activated prokallikrein at both pH 5.5 and 7.5. Prokallikrein was also activated by a highly enriched liver Golgi membrane fraction and by an endoplasmic reticulum preparation, but the Golgi preparation was 38-fold more active. The activation was blocked significantly by inhibitors of serine proteases and less by cysteine protease inhibitors.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7490146 TI - Vasoactive diadenosine polyphosphates promote growth of cultured renal mesangial cells. AB - Diadenosine polyphosphates (diadenosine triphosphate, Ap3A; diadenosine tetraphosphate, Ap4A; diadenosine pentaphosphate, Ap5A; diadenosine hexaphosphate, Ap6A) are potent vasoactive molecules stored and released by platelets. We examined whether these dinucleotides might contribute to the glomerular inflammatory response by stimulating the proliferation of mesangial cells. In cultured rat mesangial cells all four tested dinucleotides (10 to 100 mumol/L) significantly stimulated DNA synthesis as measured by [3H]thymidine uptake at 48 hours (x-fold increase compared with unstimulated control cells: Ap3A, 1.5; Ap4A, 1.8; Ap5A, 1.6; Ap6A, 1.6). In combination with the platelet products platelet-derived growth factor, epidermal growth factor, and serotonin, the dinucleotides synergistically increased DNA synthesis. Dinucleotides by themselves increased cell counts by 23% to 43% at day 2 and augmented mesangial cell growth induced by platelet-derived growth factor, epidermal growth factor, and serotonin. Furthermore, dinucleotides (100 mumol/L) rapidly induced a modest increase in expression of the early growth response gene Egr-1 at 30 minutes (x fold increase over baseline control: Ap3A, 1.9; Ap4A, 2.8; Ap5A, 2.2; Ap6A, 2.1). We found that extracellular Ap4A was metabolized by mesangial cell ectoenzymes to mononucleotides and adenosine, which also have been shown to be mitogenic for mesangial cells. The combination of Ap4A with mononucleotides or adenosine failed to cause additive stimulation of DNA synthesis in mesangial cells. We conclude that diadenosine polyphosphates stimulate proliferation of cultured mesangial cells and augment mesangial cell growth induced by other mitogens released from platelets. Different molecular mechanisms may be involved in dinucleotide-induced mitogenesis of mesangial cells. Direct effects of dinucleotides on cultured mesangial cells. Direct effects of dinucleotides on cultured mesangial cells appear to play a role because dinucleotides rapidly caused activation of Egr 1.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7490147 TI - Effect of hypothalamic-hypophysary inhibitory factor on mesangial cell activation. AB - We examined the effect of a sodium pump inhibitor isolated from bovine hypothalamus and pituitary tissues on contraction, proliferation, and calcium mobilization in primary cultures of rat mesangial cells. Hypothalamic-hypophysary inhibitory factor (HHIF) inhibited rubidium uptake in a concentration-dependent manner (0.2 U/mL: 56.8 +/- 6.3% inhibition). It also induced a concentration- and time-dependent decrease in planar cell surface area. Maximal contraction (25 +/- 5% reduction in cell size) was reached at 60 minutes with a concentration of 0.2 U/mL. This effect was inhibited by both verapamil and TMB-8 (10(-5) mol/L). HHIF was also observed to increase DNA synthesis (0.2 U/mL: 4361 +/- 168 versus 2129 +/- 162 cpm per well under control conditions) and cell proliferation (0.2 U/mL: 52,290 +/- 1931 versus 10,512 +/- 121 cells per well under control conditions). Both effects were also inhibited by verapamil and TMB-8. Moreover, HHIF induced the expression of immediate early genes c-fos and c-jun mRNA. HHIF-induced effects were accompanied by an increase in cytosolic free calcium (203 +/- 58 versus 101 +/- 2 nmol/L under control conditions), which was inhibited by verapamil and TMB-8. In summary, HHIF induces mesangial cell contraction and proliferation; these effects seem to be mediated by an increase in cytosolic free calcium levels. PMID- 7490148 TI - A consensus view on the technique of ambulatory blood pressure monitoring. The Fourth International Consensus Conference on 24-Hour Ambulatory Blood Pressure Monitoring. AB - This review, based on the Fourth International Consensus Conference on Ambulatory Blood Pressure Monitoring (Leuven, Belgium, 1994), deals with the technical aspects of ambulatory blood pressure monitoring. Ambulatory blood pressure monitoring by noninvasive intermittent techniques is widely used despite artifacts due to cuff size, movement, body position, short-term blood pressure variability, and interference with sleep. The performance of the currently available monitors under truly ambulatory conditions and during exercise remains a matter of debate, as are the procedures required to validate portable monitors under these circumstances. There is general agreement that whenever a monitor is to be used in special populations, such as older subjects or pregnant women, or in special conditions, such as exercise, a specific demonstration of its accuracy in these defined subgroups or conditions is warranted. Whether the auscultatory or oscillometric method is preferred remains controversial because each technique has specific advantages and disadvantages and because both can provide accurate results. Most experts in the field strongly believe that manufacturers should disclose the algorithms of their devices and that they should specify all changes made to the hardware and software of a previously validated monitor. Finally, the development of the volume-clamp method, which makes continuous noninvasive registration of blood pressure at the finger possible in both stationary and ambulatory conditions, opens new perspectives in research, in particular in relation to short-term blood pressure variability. PMID- 7490149 TI - Heritability of conventional and ambulatory blood pressures. A study in twins. AB - Conventional and 24-hour ambulatory blood pressures were measured in 26 pairs of monozygotic twins and 27 pairs of dizygotic twins, all male, ages 18 to 38 years, to determine the heritability of blood pressure measured under various conditions. Conventional pressure was the average of three well-standardized measurements in the supine position, and ambulatory pressure was recorded during the subjects' normal activities by use of the SpaceLabs 90202 device. Heritability was assessed by classic methods and by model fitting and path analysis. In the latter approach, the percent genetic variance was 70% for mean 24-hour systolic pressure and 73% for diastolic pressure, which was similar to the results for the conventional pressures (64% and 73%, respectively). During the night, these estimates were 72% and 51% for systolic and diastolic pressures, respectively, and also the average pressures of the total awake daytime period were under partial genetic control (63% and 55%, respectively). The remaining variances could be attributed primarily to unique environmental influences. However, shared and nonshared environmental factors were predominant for the pressures during a fixed 6-hour afternoon period. We conclude that the heritability of blood pressure is relatively high in young adult healthy men, for standardized conventional pressure and the average 24-hour pressure. Genetic variance is somewhat higher for the asleep pressure than for the awake systolic pressure. PMID- 7490150 TI - Endothelin in the kidney in malignant phase hypertension. AB - A role for endothelin in malignant phase hypertension has been suggested on the basis of reported increases of circulating plasma immunoreactive endothelins in animal models. Recently, a hypertensive rat model that exhibits a genetically determined tendency for developing spontaneous onset malignant hypertension has been described. Expression of the three genes endothelin-1, endothelin-2, and endothelin-3 was quantified in the kidney by specific RNase protection assays in rats with established malignant hypertension, in rats with benign hypertension with and without a genetic susceptibility to malignant hypertension, and in normotensive Sprague-Dawley rats. Endothelin-1 mRNA levels were significantly elevated in the group with malignant hypertension compared with the other three groups. For determination of whether endothelin-1-mediated effects were crucial in the transition from benign to malignant phase hypertension, an oral nonspecific combined endothelin-A and endothelin-B receptor antagonist (bosentan) was given to hypertensive rats susceptible to malignant hypertension. No hypotensive effects were observed, and no significant difference in the incidence of malignant hypertension was observed between treated and control groups. In conclusion, although increased endothelin-1 mRNA expression was found in kidney tissue from rats developing malignant hypertension, blockade of endothelin-1 mediated effects did not prevent the transition from benign phase hypertension. Hence, increased renal endothelin-1 expression in this model of malignant hypertension does not appear to have a causative role and may simply reflect cellular damage and ischemia. PMID- 7490151 TI - Effects of an endothelin receptor antagonist in rats with cyclosporine-induced hypertension. AB - Cyclosporine, a potent immunosuppressant, is associated with the development of hypertension and nephrotoxicity. We have previously shown that endothelin release from the arteries is increased in rats with cyclosporine-induced hypertension. We conducted the present study to determine whether the specific endothelin type A (ETA) receptor antagonist FR 139317 prevents cyclosporine-induced hypertension and whether cyclosporine increases ETA receptor mRNA in blood vessels. Cyclosporine (25 mg/kg per day) given for 4 weeks increased blood pressure from 98 +/- 12 to 156 +/- 14 mm Hg; this increase was blunted by coadministration of 10 mg/kg per day FR 139317 (ie, blood pressure was 138 +/- 14 mm Hg) in Wistar Kyoto rats. Cyclosporine induced greater vasoconstrictor responses to norepinephrine and angiotensin II in isolated mesenteric arteries. FR 139317 normalized the vasoconstrictor responses to angiotensin II and norepinephrine. Cyclosporine (25 mg/kg per day) given for 4 weeks increased ETA receptor mRNA expression in the rat aorta and mesenteric artery (170% and 176%, respectively). Little change was observed in ETB receptor mRNA. These results indicate that cyclosporine may increase blood pressure by increasing not only endothelin production but also ETA receptor in the vasculature. The specific ETA receptor antagonist FR 139317 may prevent the hypertension induced by cyclosporine. PMID- 7490153 TI - The trough-to-peak ratio as an instrument to evaluate antihypertensive drugs. The APTH Investigators. Ambulatory Blood Pressure and Treatment of Hypertension Trial. AB - The U.S. Food and Drug Administration designed the trough-to-peak ratio as an instrument for the evaluation of long-acting antihypertensive drugs, but the ratios are usually reported without accounting for interindividual variability. This study investigated how the trough-to-peak ratio would be affected by interindividual and intraindividual variability and by smoothing of the diurnal blood pressure profiles. The ambulatory blood pressure was recorded on placebo in 143 hypertensive patients (diastolic pressure on conventional measurement > 95 mm Hg). After 2 months, the recordings were repeated on 10 mg (n = 66) or 20 mg (n = 77) lisinopril given once daily between 7 and 11 PM. The baseline-adjusted trough to-peak ratios were determined from diurnal blood pressure profiles with 1-hour precision. Lisinopril reduced (+/- SD) the 24-hour pressure by 16 +/- 17 mm Hg for systolic and 10 +/- 10 mm Hg for diastolic (P < .001). According to the usual approach, disregarding interindividual variability, the trough-to-peak ratio was 0.72 for systolic pressure and 0.67 for diastolic pressure. In the 143 patients the ratios were not normally distributed. They were the same on both lisinopril doses. When interindividual variability was accounted for, the median trough-to peak ratio was 0.34 (P5 to P95 interval, -0.46 to 0.87) for systolic pressure and 0.26 (-0.44 to 0.84) for diastolic pressure. In 66 patients examined twice on 10 mg lisinopril at a median interval of 32 days, the trough-to-peak ratios were characterized by large intraindividual variability.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7490152 TI - Contrasting effect of antihypertensive treatment on the renal response to L arginine. AB - We assessed the renal hemodynamic response to L-arginine infusion (30 g within 60 minutes) in normotensive subjects, patients with never-treated essential hypertension, and hypertensive patients controlled by long-term (more than 2 years) treatment with or without an angiotensin-converting enzyme inhibitor. The renal vasodilator response to L-arginine observed in normotensive subjects (15 +/ 4% increase in effective renal plasma flow) was abolished in untreated hypertensive patients and restored only in the group treated by angiotensin converting enzyme inhibition. In the whole population a positive correlation between the change in effective renal plasma flow and the change in urinary cGMP was obtained. It is suggested that abnormalities of the renal nitric oxide pathway not corrected by increased availability of L-arginine and reversible only on long-term treatment by angiotensin-converting enzyme inhibition may underlie the abnormal renal resistance observed in essential hypertension. PMID- 7490154 TI - Combinations of potassium, calcium, and magnesium supplements in hypertension. AB - Dietary intakes of potassium, calcium, and magnesium have each been reported to lower blood pressure, but the extent of blood pressure reduction in epidemiological studies and clinical trials has tended to be small and inconsistent. We hypothesized that combinations of these mineral supplements would lower blood pressure and that the reductions would be greater than that usually reported in studies of each cation alone. One hundred twenty-five patients 82 men and 43 women) with untreated mild or borderline hypertension were randomly assigned to daily treatment with one of the following four regimens: 60 mmol potassium and 25 mmol (1000 mg) calcium, 60 mmol potassium and 15 mmol (360 mg) magnesium, calcium and magnesium, or placebo. Standardized clinic blood pressure measurements were obtained on 3 days at baseline and after 3 and 6 months of treatment. At baseline, systolic and diastolic blood pressures (mean +/ SD) were 139 +/- 12 and 90 +/- 4 mm Hg, respectively, and dietary intakes of potassium, calcium, and magnesium were 77 +/- 32, 19 +/- 13, and 12 +/- 52 mmol/d, respectively. The mean differences (with 95% confidence intervals) of the changes in systolic and diastolic blood pressures between the treatment and placebo groups were not significant: -0.7 (-4.3 to +2.9) and -0.4 (-2.9 to +2.1) for potassium and calcium, -1.3 (-4.4 to +1.8) and 0.4 (-2.5 to +3.3) for potassium and magnesium, and +2.1 (-1.8 to +6.0) and +2.2 (-1.0 to +5.4) for calcium and magnesium. In conclusion, this trial provides little evidence of an important role of combinations of cation supplements in the treatment of mild or borderline hypertension. PMID- 7490155 TI - Chronic inhibition of nitric oxide synthesis causes coronary microvascular remodeling in rats. AB - The aim of the present study was to investigate the effects of long-term blockade of nitric oxide synthesis with the L-arginine analogue N omega-nitro-L-arginine methyl ester (L-NAME) for 8 weeks on coronary vascular and myocardial structural changes. Four groups of Wistar-Kyoto rats were studied: those with no treatment, those treated with L-NAME 1 g/L (3.7 mmol/L in drinking water), those treated with L-NAME 0.1 g/L (0.37 mmol/L in drinking water), and those treated with L NAME 1.0 g/L and hydralazine 120 mg/L (0.6 mmol/L in drinking water). After 8 weeks, the heart was excised, and the degrees of structural changes in coronary arteries (wall-to-lumen ratio and perivascular fibrosis), myocardial fibrosis, and myocyte size were quantified by an image analyzer. Chronic inhibition of nitric oxide synthesis increased arterial pressure compared with control animals. Chronic inhibition of nitric oxide synthesis caused significant microvascular remodeling (increased wall-to-lumen ratio and perivascular fibrosis). Cardiac hypertrophy was also observed after chronic inhibition of nitric oxide synthesis. Coadministration of hydralazine prevented arterial hypertension but did not affect microvascular remodeling and cardiac hypertrophy induced by the chronic inhibition of nitric oxide synthesis. In addition, chronic inhibition of nitric oxide synthesis caused scattered lesions of myocardial fibrosis, which was significantly attenuated by cotreatment with hydralazine. These results suggest that long-term blockade of nitric oxide synthesis caused coronary microvascular remodeling and cardiac hypertrophy in rats in vivo by a mechanism other than arterial hypertension. In contrast, arterial hypertension contributed to the development of myocardial fibrosis induced by long-term blockade of nitric oxide synthesis. PMID- 7490156 TI - Aortic stiffness and left ventricular mass in a rat model of isolated systolic hypertension. AB - We tested whether cardiac mass can be related to decreased aortic stiffness in an original rat model of isolated systolic hypertension. Increased aortic stiffness was produced by calcium overload of elastic arteries after vitamin D3 plus nicotine treatment. Half of the animals were chronically treated with the angiotensin-converting enzyme inhibitor perindopril (1 mg/kg per day PO). Rats were pithed, and lower body vascular resistance was measured. Blood pressure was then increased by phenylephrine infusion, and carotidofemoral pulse wave velocity was measured. This value together with those for thoracic aorta internal diameter and medial thickness (determined after in situ fixation and histomorphometry) were used to calculate elastic modulus. Vitamin D3 plus nicotine treatment produced parallel increases in cardiac mass and elastic modulus, with a significant correlation between the two. There was no significant change in resistance. Treatment with perindopril reversed the changes in cardiac mass and elastic modulus but had no effect on resistance after calcium overload of the elastic arteries. In this model of isolated systolic hypertension, we showed that cardiac mass is related to arterial elasticity. PMID- 7490157 TI - Biological determinants of aldosterone-induced cardiac fibrosis in rats. AB - To determine the events leading to cardiac fibrosis in aldosterone-salt hypertensive rats, we studied protein and mRNA accumulation of procollagens I and III for 60 days. After 3 and 7 days of treatment systolic pressure was normal, and no histological or biochemical changes were seen in rat hearts. At day 15 arterial pressure was raised (+40%) and left ventricular hypertrophy was +15%. Cardiac examination after hemalun-eosin staining and immunolabeling with anticollagen I and III antibodies showed no structural alterations, but an 83% increase in right ventricular type III procollagen mRNA levels was found. At 30 and 60 days we found progressive cardiac fibrosis, with inflammatory cells, myocyte necrosis, and elevation of both types I and III procollagen mRNA levels in both ventricles. To determine whether aldosterone had effects on Na,K-ATPase that might lead to ionic disturbances and induce myocyte necrosis, we studied the major cardiac Na,K-ATPase isoform genes. Although Na,K-ATPase alpha 1- and beta 1 subunit mRNA levels were elevated in kidney at day 1, neither of these cardiac transcripts nor the specific alpha 2 isoform was altered between 1 and 15 days. These results show that accumulation of procollagen mRNAs occurs before collagen deposition. Cardiac alterations are late and not preceded by changes in Na,K ATPase cardiac gene expression, precluding a direct modulation of cardiac collagen synthesis and Na,K-ATPase by aldosterone. PMID- 7490158 TI - Gender differences in left ventricular growth. AB - Because the number of human cardiac myocytes is determined in infancy, subsequent increases in left ventricular (LV) muscle mass reflect cellular enlargement (hypertrophy). To determine whether the greater LV mass in adult men than in women reflects sex differences that are present throughout development or disproportionate LV growth during puberty in men, we compared echocardiographic LV mass in 333 female and 278 male normal-weight, normotensive subjects from 4 months to 70 years of age. Only a small sex difference in LV mass (mean = 6%) existed before age 12 years, whereas in all older-age strata LV mass in men was 25% to 38% greater than that in women (P < .02 to P < .0001). The divergence in LV mass between male and female adolescents closely paralleled differences in height and weight and was due to proportional increases in LV chamber dimension and wall thickness in males (with no sex difference in relative wall thickness). LV mass grew less rapidly from infancy through childhood than did body size, assessed by body weight or height, yielding a reduction of LV mass/body size ratios up to puberty, which was followed by gradual increases during adulthood. Indexation of LV mass by body weight or height but not by body surface area or height markedly reduced the sex differences in LV mass/body size ratios from puberty through the seventh decade of life. Thus, before puberty, LV mass is only modestly higher in boys than in girls.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7490159 TI - Radial artery compliance in young, obese, normotensive subjects. AB - Obesity is characterized by a number of cardiovascular alterations, and whether these alterations involve arterial compliance is unknown. In 12 young, obese, normotensive subjects (age, 23.9 +/- 1.3 years; mean +/- SEM) and 12 age- and sex matched lean control subjects we measured blood pressure, radial artery diameter, and radial artery compliance continuously over the systodiastolic pressure range with a Finapres device and recently developed echo-tracking device. Measurements were obtained at baseline and after prolonged ischemia, that is, when diameter and compliance are increased. Blood pressure values were normal in both groups (obese subjects: 109.2 +/- 4.9/68.2 +/- 2.7 mm Hg; lean control subjects: 108.2 +/- 4.1/60.7 +/- 3.8 mm Hg), but in addition to a marked increase in body mass index (38.5 +/- 0.8 versus 23.1 +/- 0.9 kg/m2, P < .01), obese subjects showed a slight and nonsignificant increase in heart rate (71.1 +/- 3.2 versus 66.7 +/- 3.3 beats per minute, P = NS), increases in left ventricular wall thickness and left ventricular mass index (121.5 +/- 4.8 versus 103.4 +/- 3.3 kg/m2, P < .01), no changes in plasma renin activity and plasma norepinephrine (compared with normal values), and a marked reduction in total body glucose uptake (glucose clamp technique). Obese subjects showed radial artery diameter and compliance values that were greater than those seen in control subjects throughout the systodiastolic pressure range. The differences were 13% (P < .05) and 96% (P < .01), respectively, and both diameter and compliance remained higher in obese than lean subjects after forearm ischemia. In obese and lean subjects baseline radial artery diameter values correlated highly with body weight, body surface area, and body mass index.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7490160 TI - Regional hemodynamic effects of the AT1 receptor antagonist CV-11974 in conscious renal hypertensive rats. AB - Angiotensin II subtype 1 (AT1) receptor antagonists reduce mean arterial pressure in various experimental models of hypertension, including two-kidney, one clip (2K1C) renal hypertension. However, the regional hemodynamic mechanisms underlying the hypotensive effect of AT1 receptor antagonists in 2K1C rats under dynamic conditions have not been documented. Therefore, in the present study we determined the hemodynamic profile of the AT1 receptor antagonist CV-11974 in conscious 2K1C rats and sham-operated control rats. Approximately 4 weeks after clipping, rats underwent a further two-stage operation for implantation of Doppler flow probes on the contralateral (left) renal artery, superior mesenteric artery, and distal aorta as well as for the implantation of intravascular catheters. At least 24 hours after the last operation continuous recordings were made of mean arterial pressure; heart rate; and renal, mesenteric, and hindquarters flows and conductances (Doppler shift/mean arterial pressure) in response to three doses of CV-11974 (0.01, 0.1, and 1.0 mg/kg i.v.). CV-11974 caused a small hypotensive effect (decrease of approximately 15 mm Hg) in the sham group, but regional flows and vascular conductances did not change. By contrast, in 2K1C rats CV-11974 caused dose-dependent hypotension that was maximal (-19 +/- 6, -41 +/- 4, and -51 +/- 8 mm Hg, respectively) after 6 hours. These changes were associated with generalized vasodilatation (increased conductance) in all three vascular beds, although there were subtle differences with the different CV-11974 doses.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7490162 TI - Building up a computerized follow-up register and information system for cervical cytology. AB - It is well known that cervical intraepithelial neoplasia (CIN) and human papilloma virus (HPV) infection are closely related to cervical cancer. The cervical cytology examination (pap-test), followed by a colposcopic examination and a biopsy are the common tools for the early detection and the establishment of diagnosis of the cervical precancerous lesions. The monitoring and surveillance of those precancerous lesions is essential for the prevention of cervical cancer and for effective patient management. Nowadays, the long-term care of patients with a chronic and/or serious disease is feasible with the use of computerized follow-up registers or through more complex information systems. Nevertheless, manual recording is still the predominant method of data collection in 'Alexandra' University Hospital and this is the case in most health services in Greece due to scarcity of resources. A 'circulating' outpatient card, where information on gynecologic cytology, colposcopy and biopsy examinations is recorded, was the basis for the development of a computerised follow-up register and information system for cervical cytology in the Department of Cytopathology. PMID- 7490163 TI - Automatic detection of auditory brainstem responses using feature vectors. AB - The efficiency of several methods for the automatic detection of near threshold auditory brainstem responses (ABRs), based on feature vectors, was assessed on a sample of 374 recordings obtained from neonates. Features are quantitative descriptors of different aspects of the response commonly taken into consideration by expert ABR judges, such as: similarity with a template, reliability of the response, and intrinsic morphological characteristics. The classification methods used were: comparisons of individual features with a threshold, linear discriminant functions based on combination of features, artificial neural networks (ANNs) based on the raw ABR, and ANNs based on combinations of features. The network used was a modified version of the Fuzzy ART Map model. The accuracy of the classification into normal and abnormal (as previously scored by a panel of experts) was assessed with methods from signal detection theory. Our results showed that the approaches based on feature vectors (discriminant function and ANN) performed more efficiently than the ANN with raw data, or the individual features. PMID- 7490161 TI - Cloning, characterization, and genetic mapping of the rat type 2 angiotensin II receptor gene. AB - The renin-angiotensin system plays an important role in blood pressure homeostasis, but the contribution of the type 2 angiotensin II receptor (AT2R) is still unclear. The reports that the AT2R gene has been mapped to the X chromosome in human and rat and the previous report of a gene, Bp3, on the X chromosome responsible for an increase in blood pressure have suggested that the rat AT2R gene (Agtr2) could be this gene. To elucidate whether Agtr2 is Bp3, Agtr2 was cloned. A simple sequence repeat in the 3'-flanking region of this gene was identified and used as a genetic marker to map Agtr2 to the X chromosome at 18.1 cM distal to the androgen receptor locus. This map position is outside the confidence interval reported for Bp3, demonstrating that Agtr2 cannot be Bp3. However, these data will enhance the research into the AT2R biology as well as the study of the X chromosome. PMID- 7490164 TI - An expectation maximization reconstruction algorithm for emission tomography with non-uniform entropy prior. AB - A Bayesian image reconstruction algorithm is proposed for emission tomography. It incorporates the Poisson nature of the noise in the projection data and uses a non-uniform entropy as an a priori probability distribution of the image in a maximum a posteriori (MAP) approach. The expectation maximization (EM) method was applied to find the MAP estimator. The Newton-Raphson numerical method whose convergence and positive solutions are proven, was used to solve the EM problem. The prior mean at iteration k was determined by smoothing the image obtained at iteration k-1. Comparisons between the ML and the MAP algorithm were carried out with a numerical phantom that contains a narrow valley region. The ML solution after 50 iterations was chosen as the initial solution for the MAP algorithm, since the global performance of the ML algorithm deteriorates with increasing number of iterations while its local performance in the valley region is always improving. The resulting algorithm is a compromise between ML who has the best local performance in the valley region and the MAP who has the best global performance. PMID- 7490165 TI - Wavelet analysis of high-resolution ECGs in post-infarction patients: role of the basic wavelet and of the analyzed lead. AB - Wavelet analysis provides a fruitful alternative to standard techniques for the detection of fractionated potentials in signal averaged high-resolution (SA-HR) ECGs. In this study, an attempt is made to optimize the discrimination of post infarction patients prone to ventricular tachycardia (VT), using wavelet analysis. Optimization is based on the choice of the ECG leads or lead combinations to be analyzed, and on the analyzing wavelet to be computed. A set of 40 post-infarction patients (20 patients with VT and 20 patients without any arrhythmia) is analyzed. Individual leads and lead combinations of the SA-HR ECGs are processed using a multiparametric algorithm, based on coherent detection of aligned local maxima of the wavelet transform. Seven basic wavelets are tested: the Morlet's wavelet, and the six first derivatives of a Gaussian function. The first derivative of a Gaussian function provides poor results, and is discarded. All other wavelets prove to perform equivalent classification. A vector magnitude computed from the wavelet transforms of the three SA-HR ECGs achieves better results than individual leads. An optimized risk stratification algorithm leads to 90% sensitivity and 100% specificity in the 40 patients learning set. PMID- 7490168 TI - Extraction of discriminant features in post-cardiosurgical intensive care units. AB - A linear transformation, based on the Karhunen-Loeve expansion, is applied to 13 physiological variables, measured in 200 surgical patients, in order to extract a limited number of features well representative of the differences between normal and high-risk classes of subjects. This transformation may be considered as a mapping from the primitive 13-dimensional space to a lower dimensional one, without severely reducing class separability. The efficacy of both transformed and primitive variables in the separation of normal and high-risk subjects is compared using the error probability, i.e. the probability that a patient is assigned to the wrong class. In particular, its upper bound is evaluated through the Kullback divergence and its estimate is computed, from the available samples, by applying a quadratic classifier. The results obtained show that only two transformed variables are able to present a divergence better than the most effective set of eight primitive variables. In agreement with the divergence criterion, the classifier provides a recognition error lower than 5% and greater than 13% when using the two best transformed and the two best primitive variables, respectively. Even though the new variables do not have a direct physiological meaning, this limitation has been partially overcome by calculating the correlation matrix between transformed and primitive variables. The results presented show that the first two transformed variables are strongly related to the most discriminant primitive ones (i.e. cardiac index, oxygen delivery and arterio-venous oxygen difference). In conclusion, the transformation of variables proposed appears to be extremely attractive for practical applications, since it allows recognition systems to be designed which exhibit both high performance and great simplicity. PMID- 7490167 TI - Mathematical representation of organ growth in the human embryo/fetus. AB - During human pregnancy, there is a huge increase in the total weight of the embryo/fetus from conception to term. The total growth, which is the summation of growth of the various organs and tissues that make up the organism, was analyzed in a previous paper and fitted to the Gompertz equation for growth. In the present study, allometry, the quantitative representation of the consequence of size, was utilized to describe the correlation of individual fetal organ/tissue weights with the total fetal weight. The organ/tissue weight and the total fetal weight data used in the analyses were pooled from various sources that provided data ranging from 25 days to 300 days post-conception. Allometric equations are presented for 16 embryo/fetal organs and tissues. The standard allometric equation gave adequate fits for embryo/fetal adrenal, bone, bone marrow, brain, heart, liver, pancreas, plasma, skeletal muscle, extracellular water, thymus and thyroid; but it was necessary to use a quadratic form of the allometric equation for embryo/fetal fat, kidney, lung and spleen. Parameters were also calculated for crown-to-rump and crown-to-heels for fetal lengths that occur during pregnancy. PMID- 7490166 TI - Comparison of inference results of two otoneurological expert systems. AB - In this paper, two different otoneurological expert systems, Vertigo and One, the latter developed by us, are considered. The expert systems are evaluated as regards their correctness in reasoning diagnoses. In the light of our data collected from randomly selected test patients, One, being a newer technique, is more effective, since it could infer more cases than vertigo did. All the data was also evaluated and diagnosed by otoneurological specialists, independently of the expert systems, to guarantee objectivity in evaluation of the results of the expert systems. PMID- 7490169 TI - The effects of bifemelane hydrochloride on depressive illness of the elderly. AB - The therapeutic efficacy, utility and safety of bifemelane hydrochloride were studied in 52 elderly depressive patients. The drug was administered as a tablet containing 50 mg orally three times daily for 8 consecutive weeks. The final global improvement rating and global utility rating were respectively 80.8 and 73.1 percent for all patients. The improvement rates on the Hamilton depression rating scale (HAM-D) were more than 60% for depressed mood, guilt, suicide, middle insomnia, delayed insomnia, psychotic anxiety, gastro-intestinal symptom, hypochondriasis, depersonalization and derealization. The rates regarding global symptoms evaluated by the Psychoneurotic rating scale for doctor's use were more than 60% for tension, agitation, irritability and excitement, phobia, depression, hypochondria and nocturnal delirium in psychotic symptoms, and insomnia in addition to palpitation in somatic symptoms. A significant decrease was also observed in the symptoms covered by the Self-rating depression scale of Zung after treatment with this drug. There were no instances of side-effects, nor any abnormalities in laboratory tests, encountered throughout the trial. Therefore, bifemelane hydrochloride is of value for the treatment of geriatric depression. PMID- 7490170 TI - Resistance patterns to beta-lactams and quinolones in clinical isolates of bacteria from Cuban hospitals. AB - The resistance patterns to 26 beta-lactams and 8 quinolones of clinical isolates from Cuban hospitals were evaluated using the disk susceptibility test, according to the NCCLS guidelines (1992). The genera studied were Escherichia sp (320), Enterobacter sp (10), Klebsiella sp (90), Proteus sp (10), Pseudomonas sp (90), Serratia sp (20), and Staphylococcus sp (80). Higher resistance to beta-lactams was observed in the genera Pseudomonas, Escherichia and Klebsiella. For fluoroquinolones we found no significant resistance, with the exception of the genus Klebsiella. The most effective antibiotics were cephalosporins of the second and third generations, fluoroquinolones, and non-classical beta-lactams (cephamycins, moxalactam and monobactams). On the contrary, a pronounced resistance was found to penicillin, oxacillin, ticarcillin, ampicillin, methicillin, nalidixic acid and cinoxacin. These resistance patterns correspond to the high consumption of these antibiotics throughout the country. PMID- 7490171 TI - Effects of cardiopulmonary bypass on teicoplanin serum disposition. AB - The time course of teicoplanin (T) serum concentrations was determined in 6 patients who had undergone cardiopulmonary bypass (CPB). The drug was given i.v. (12 mg/kg) 85-140 min before starting CPB. Serum concentrations of T were measured with an automated fluorescence polarization immunoassay, at appropriate times before, during and after CPB (total sampling interval: 12 h). Five min after initiating CPB, T serum concentrations decreased, on average, by 29% and remained less than the expected values (values extrapolated from the decay curve, measured excluding the CPB period) over the subsequent 60 min. When CPB was discontinued, the T serum concentrations rebounded to the expected values within 5 min. The mean area under the curve (AUC) during CPB was significantly lower than the mean extrapolated AUC. It was concluded that CPB reversibly reduced T serum concentrations, probably due to drug redistribution. Nevertheless, T serum levels were always above the mean inhibitory concentration (MIC) of the most common pathogenic organisms associated with CPB surgery infections. PMID- 7490172 TI - Intra-articular somatostatin 14 reduces synovial thickness in rheumatoid arthritis: an ultrasonographic study. AB - Sixteen patients with RA (3 males, 13 females), diagnosed according to RA revised criteria, were selected and entered the study. They underwent six intra-articular injections of 750 mcg of SST14 at 15-day intervals. The thickness of the synovial membrane (SM) was measured with a 5-MHz linear sound with longitudinal and transversal scanning carried out on the upper patellar cavity. The contralateral knee was also assessed together with the injected knee in order to ascertain any systemic effect of the drug. A significant reduction of SM thickness was observed already at the first control (T3) in 14 out of 16 patients. At the 5th and 6th injections (T5 and T6) the reduction was still significant but to a lower extent. In 8 out of 16 cases a reduction of SM thickness was observed in the contralateral knee. Analysis of these data clearly shows that the intra-articular injection of SST14 is able to reduce the thickness of SM in patients with RA, and indicates that SST14 may directly reduce synovitis. This particularity has been detected in our work with a non-invasive technique such as the joint ultra-sound (US). In conclusion, our work confirms the efficacy of SST14 in the control of RA synovial hypertrophy and the reliability of US technique in the measurement of SM thickness. PMID- 7490173 TI - L-acetylcarnitine as a new therapeutic approach for peripheral neuropathies with pain. AB - With ST200 as the commercial source of L-acetylcarnitine hydrochloride, 94 patients were enrolled in this study; 31 were assigned to placebo, 31 to ST200 at 0.5 g/die and 32 to ST200 at 1 g/die, the i.m. treatments being injected daily for 15 consecutive days. In general, concerning the efficacy assessment, the administration of ST200 at 1 g/die appeared to be better than ST200 at 0.5 g/die when compared with the placebo administration. Statistically significant differences were revealed by the comparison of ST200 at 1 g/die to placebo, for the following variables: a) total motility as rated at the end of the 15-day study and confirmed by intention-to-treat analysis, b) visual analogue scale for all the patients having observation at day 15, and c) the objective and subjective judgements on efficacy. Safety and tolerability were good over the entire course of the study. PMID- 7490174 TI - Posterior uveitis in the acquired immunodeficiency syndrome. PMID- 7490175 TI - Human T-lymphotropic virus type 1 and its ocular manifestations. PMID- 7490176 TI - Cataract extraction in patients with chronic posterior uveitis. PMID- 7490177 TI - Ocular toxoplasmosis: toxoplasmic retinochoroiditis. PMID- 7490178 TI - Sympathetic ophthalmia. PMID- 7490179 TI - Behcet's disease. PMID- 7490180 TI - Vogt-Koyanagi-Harada syndrome. PMID- 7490181 TI - Serpiginous (geographical) choroiditis. PMID- 7490182 TI - Acute posterior multifocal placoid pigment epitheliopathy. PMID- 7490183 TI - Double contrast upper gastrointestinal barium examination with biopsy versus endoscopy with biopsy in dyspeptic patients. AB - RATIONALE AND OBJECTIVES: The financial restrictions of the managed care environment require reconsideration of the barium upper gastrointestinal examination as a diagnostic tool for gastritis patients. However, a greater sensitivity and specificity for gastritis is needed. A prospective study was performed comparing barium examinations with gastric biopsies to endoscopy with biopsy. METHODS: Forty adult patients underwent upper gastrointestinal barium examination with gastric biopsies obtained under fluoroscopy through a nasogastric tube. Twenty-seven patients gave consent for subsequent endoscopy with biopsy. Both sets of biopsies were compared, as were the interpretations of the radiographs and visual appearances. RESULTS: For barium examinations with gastric biopsies, sensitivity for gastritis was 94% and specificity was 100%, using endoscopic biopsies as the gold standard. CONCLUSIONS: In addition to endoscopy with biopsy, the upper gastrointestinal barium examination with biopsy is another option of sufficient sensitivity and specificity for consideration by clinicians in their workup of patients with gastritis. PMID- 7490184 TI - Guidelines for the assessment of new diagnostic tests. AB - RATIONALE AND OBJECTIVES: Because new diagnostic tests become available rapidly, the authors determined a need for proper assessment of tests before their implementation in clinical practice. Three factors are of pivotal importance: the selection of the proper study population, the determination of the diagnostic power including its related statistical analysis, and the relation of the new test to current diagnostic tools. Patients suspected of having a disease are those who would benefit from the application of a new test. Therefore, only those patients need be involved in the assessment study. METHODS: Summary measures of diagnostic power other than sensitivity and specificity are advocated because these conventional measures depend on cutoff points and are susceptible to selection bias. The relation between the new test and existing diagnostic tools must be established to determine if the new test contributes to the diagnostic process. RESULTS AND CONCLUSION: To avoid waste of effort and money, the authors suggest a prudent assessment approach in phases. Whereas the initial challenge consists of selection of an adequate patient population, subsequently all determinants of diseases (signs, symptoms, comorbidity, and other diagnostic factors) and factors influencing the decision to use a test (patient burden and cost) are considered. PMID- 7490185 TI - Use of fibrinolytic agents to coat wire implants to decrease infection. An animal model. AB - RATIONALE AND OBJECTIVES: Infection is a serious complication of metallic prosthesis implantation and may necessitate removal of the prosthesis. This study uses an animal model to evaluate the effects of coating stainless steel wire implants with fibrinolytic agents to prevent infection after bacterial contamination. METHODS: Three types of steel wire implants were used: plain stainless steel, heparin-coated steel, and urokinase-heparin-coated steel. Wire implants were incubated in a known concentration of Staphylococcus epidermidis and placed into the subcutaneous tissues of three groups of anesthetized hamsters. The implants and surrounding tissues were excised after 1 week and submitted for quantitative cultures. RESULTS: Using 100 organisms as the upper allowable limit to categorize abscesses as noninfected, the following rates of noninfectivity were observed: group 1 (control), 0% noninfected; group 2 (heparin coated wire), 40% noninfected; and group 3 (urokinase-heparin-coated wire), 50% noninfected. The noninfectivity rates of groups 2 and 3 were significantly higher than the rate of group 1 (P < 0.001). There was no significant difference between groups 2 and 3 (P = 0.19). CONCLUSIONS: Both the heparin-coated and urokinase heparin-coated wire exhibited significantly decreased infection rates compared with uncoated wire; the heparin coating may inhibit bacterial adherence. The urokinase coating of the heparin-coated wire appears to further decrease the infection rate, but not to a statistically significant degree. PMID- 7490187 TI - Percutaneous internal maxillary arterial embolization with ethylcellulose microspheres. Results in an animal model. AB - RATIONALE AND OBJECTIVES: To investigate the use of ethylcellulose microspheres as long-term and peripheral emboli for percutaneous maxillofacial arterial embolization. METHODS: Eight mongrel dogs were selected randomly for internal maxillary artery embolization with ethylcellulose microspheres. After embolization, angiographic, microangiographic, and histologic examinations were performed. RESULTS: Ethylcellulose microspheres were trapped in the peripheral arterioles from 24 hours to 6 months after embolization. Degenerative changes of maxilla, mandible, and dental pulp occurred after the embolization of the internal maxillary artery with the microspheres. No evidence of whole or focal necrosis of the bones and surrounding soft tissues was found between 24 hours and 6 months after embolization. CONCLUSION: Ethylcellulose microspheres can be used as an alternative long-term and peripheral embolic agent, with potential for percutaneous maxillofacial arterial embolization. PMID- 7490186 TI - Calcification can shorten T2, but not T1, at magnetic resonance imaging fields. Results of a relaxometry study of calcified human meningiomas. AB - RATIONALE AND OBJECTIVES: Water content and water-proton relaxation rates are reported for fresh, histologically characterized, surgical specimens of calcified human intracranial meningiomas and compared with results for noncalcified meningiomas from an earlier study and with calcium hydroxyapatite (CaHA) suspensions to elucidate the influence of calcification on magnetic resonance imaging (MRI) signal intensity of calcified meningiomas. METHODS: The magnetic field dependence of 1/T1 of water protons (nuclear magnetic relaxation dispersion profile) and dry weights are reported for 38 calcified nonhemorrhagic and 3 hemorrhagic specimens of known histologic subtype, a subset of the 67 specimens measured earlier. Calcification was considered mild or heavy when the dry weight was within or above the range for noncalcified meningiomas. Preliminary 1/T1 profiles for pure CaHA and a single high-field 1/T2 value also are reported. RESULTS: The ranges of dry weights and of low-field 1/T1 values were twice as large for calcified as for noncalcified meningiomas. No correlation was found between low-field 1/T1 and either histologic subtype or dry weight. Mild calcification produced the highest low-field 1/T1 values; the most heavily calcified tumor had slightly increased low-field 1/T1. Calcium hydroxyapatite increases low-field 1/T1 significantly but not high-field 1/T1; high-field 1/T2 is large. For calcified hemorrhagic meningiomas, increases in both low-field and high-field 1/T1 were seen. CONCLUSION: For mild calcification, MRI signal voids result from an increased high-field 1/T2; for heavier calcification, reduced proton density (from excluded water) becomes of increasing importance. Cellular CaHA appears to brighten the signal in T1-weighted MRI in the presence of hemorrhage. PMID- 7490189 TI - Comparison of single-tracer (technetium-99m-sestamibi) and dual-tracer (thallium 201 chloride and technetium-99m-sestamibi) protocols for identification of myocardial ischemia. AB - BACKGROUND: The objective of this study was to determine if dual-tracer protocol using thallium-201 (201TI) chloride for rest and technetium-99m (99mTc)-sestamibi for stress images (TI-MIBI) detects ischemia better than single-tracer protocol using 99mTc-sestamibi for both rest and stress (MIBI) myocardial perfusion images. METHODS AND RESULTS: A total of 460 patients with suspected or known coronary artery disease (CAD) were included in the study, with 230 (50%) undergoing single-tracer MIBI protocol and 230 (50%) undergoing dual-tracer TI MIBI protocol. Patient demographics were matched, and the proportion of patients having treadmill stress or dipyridamole stress was similar between the two groups. Stress and rest single photon emission computed tomographic images were recorded in all patients and dual-tracer 201TI and 99mTc-sestamibi (TI-MIBI) perfusion images were acquired separately. The scintigraphic images were analyzed visually, and a semiquantitative method of scoring reversible ischemia was used. Patients' reversibility scores in each group were compiled (ischemia index) and compared to determine the relative ability of each protocol in identifying ischemia. CONCLUSION: The frequency of ischemia and ischemia index was found to be significantly (P < 0.05) higher in the TI-MIBI group compared with the MIBI group. For detecting myocardial ischemia or viability, the dual-tracer TI-MIBI acquisition technique appears superior to the single-tracer MIBI protocol. PMID- 7490188 TI - One-dimensional phosphorus-31 chemical shift imaging of human brain tumors. AB - RATIONALE AND OBJECTIVES: Phosphorus magnetic resonance spectroscopy has been used noninvasively to determine characteristic spectral parameters for untreated human brain tumors as a prelude to its use in clinical diagnosis. METHODS: The spectra, which reflect the relative amounts of phosphorus-containing compounds, and the pH within and surrounding the tumors, were obtained in vivo using the localization technique of one-dimensional chemical shift imaging applied with a surface coil. Phosphorus-31 chemical shift imaging was performed successfully in vivo on 9 volunteers and 27 patients with untreated brain tumors, including 7 with astrocytoma, 4 with glioblastoma, 3 with meningioma, and 11 with metastases. This study provides spectra from within and surrounding the brain tumors, and allows accountability for the heterogeneity of brain tumors by the selection of the maximum data point for each parameter. RESULTS: The ratios of resonance areas, phosphodiesters over nucleoside triphosphate (NTP), and phosphomonoesters over NTP, were found to be higher in glioblastomas (2.55 +/- 0.22, 1.06 +/- 0.09) and astrocytomas (3.04 +/- 0.36, 1.28 +/- 0.36) than in normal brain (2.00 +/- 0.32, 0.79 +/- 0.22). The ratios of areas due to inorganic phosphate and NTP, and phosphocreatine and NTP, also were higher in astrocytomas (1.16 +/- 0.40, 1.17 +/ 0.41) compared with glioblastomas (0.68 +/- 0.01, 0.88 +/- 0.19) and normal brain (0.61 +/- 0.03, 0.77 +/- 0.03). The pH of brain tumors ranged from alkaline to neutral, with meningiomas consistently having alkaline pH. CONCLUSIONS: These data show that there are statistically significant differences in the magnetic resonance parameters of the affected brain hemispheres of patients with astrocytomas, glioblastomas, meningiomas, and normal brain tissue, and underline the need for a multisite clinical trial to establish clinical criteria for diagnosis. PMID- 7490191 TI - Recurrent fibromatosis in a patient with breast carcinoma. Computed tomography findings with magnetic resonance imaging and pathologic correlation. PMID- 7490190 TI - Biodistribution of radiolabeled, formulated gadopentetate, gadoteridol, gadoterate, and gadodiamide in mice and rats. AB - RATIONALE AND OBJECTIVES: The authors studied the long-term distribution of gadolinium (Gd) in mice and rats after the administration of four commercially available magnetic resonance imaging contrast media. The goals were to determine any possible product dissociation in vivo and to evaluate the effects that product excipients had on the tissue distributions. METHODS: Gadolinium-153 (153Gd)-labeled gadopentetate (Magnevist), gadoteridol (ProHance), gadoterate (Dotarem), and gadodiamide (Omniscan) were administered intravenously to mice (0.48 mmol/kg) and rats (0.1 mmol/kg). At various times up to 14 days posttreatment, the residual 153Gd was measured in selected tissues. The tissue distributions obtained were used to make intra- and interchelate distribution evaluations and comparisons regarding tissue clearance and any possible in vivo dissociation of the Gd chelates. RESULTS: Differences were found among the chelates studied relative to the amounts of residual 153Gd present in tissues known to sequester free Gd, particularly in liver and femur at 7 and 14 days after administration, in both mice and rats. The pattern of the 153Gd distribution suggested that the linear chelates, gadopentetate and gadodiamide, dissociated in vivo resulting in more 153Gd present in bone and liver at the longer residence times than in the subjects injected with the macrocyclic chelates, gadoteridol and gadoterate. The only excipient found to affect the distribution profile was calcium(DTPA-BMA); this excipient in formulated gadodiamide decreased the amounts of residual Gd measured in whole body, bone, and liver in mice compared with levels obtained when gadodiamide was injected alone. CONCLUSIONS: The molecular feature found to be most important in differentiating the four chelates evaluated is the presence or absence of a macrocyclic structure. The Gd chelates containing this structure, gadoteridol and gadoterate, have the lowest residual Gd at long residence times in both mice and rats. The order of residual whole body Gd at 14 days (lowest to highest) was: gadoteridol integral of gadoterate < or = gadopentetate << gadodiamide. The only excipient that affected the biodistribution was found in the gadodiamide formulation where the addition of 5% calcium (DTPA-BMA) reduced residual Gd to just less than 10 times greater than that found for the macrocyclic chelates with the lowest residual Gd, gadoteridol and gadoterate. PMID- 7490192 TI - Information for all doctors. Update of success of measles/rubella immunisation campaign. PMID- 7490193 TI - Research ethics committees: principles and proposals. PMID- 7490194 TI - Coping with your coronary heart disease target. AB - The Scottish targets for coronary heart disease are examined in detail from the point of view of a Health Board. It is suggested that even though the target refers to the under 65 age group, further age/sex standardisation is needed for fair comparison. A least-squares regression technique is also recommended as a means of reducing the effect of atypical rates in baseline or target years. The targets adopted do not make it clear whether Health Boards should aim for a target in terms of the percentage fall, or should aim to be below an absolute level by the target year. The specific targets imply that resources will have to be concentrated in people of late middle age, particularly males, to the detriment of preventive activities in younger age groups. It seems necessary to emphasise that targets are not an end in themselves but merely an indicator of progress. Monitoring progress towards achieving the target could be achieved by assuming a linear fall in rate between baseline and target years, to produce annual intermediate targets. A health board's progress towards the final target can then be annually monitored by comparison of its regressed age-sex standardised rate with the intermediate target. PMID- 7490195 TI - Health promotion and the use of Gpass in Scotland. AB - OBJECTIVE: To identify how computerised practices using Gpass software (General Practice Administration System for Scotland), currently implement the new health promotion regulations. DESIGN: Postal questionnaire to all Gpass practices in Scotland. Data were gathered on types and methods of recording health promotion data, Read code selection, health education given and intended methods of data analysis. Questionnaire results were compared with data from an Electronic Questionnaire analysing actual data recorded on practice computers. RESULTS: Overall response rate: 64.6%, 94.8% of the responding practices have been approved for health promotion band three. Most practices (94.5%) use their computer for data collection, 63.6% of practices use a manual data capture form and 28.8% use computer data capture methods. Methods of collecting patient data and selection of Read codes for computer data entry are variable. Most practices use one method of data collection; a significant minority use multiple methods or more than one Read code to record the same item. The recording of health promotion on computer has increased greatly since the introduction of the new regulations: the current levels of recording are alcohol history (26.3%), blood pressure reading (57.6%), smoking (35.4%), exercise (7.1%), weight (21.4%) and height (16.4%). Most practices (94.3%) intend using Gpass for data analysis. CONCLUSION: Methods of collecting and recording health promotion data differ greatly between practices, with variable standardisation of health promotion codes and differing use of appropriate elements of the Gpass software. PMID- 7490196 TI - An audit of the clinical management of urinary tract infection in childhood. AB - Urinary tract infection (UTI), is one of the commonest bacterial diseases in childhood, and may cause significant morbidity, inconvenience and anxiety for patients and their families and result in considerable consumption of resources. Although most clinicians would agree on the general principles of management, it is recognised that considerable variation exists in their application. The aim of this study was to assess the in-patient and out-patient management practice in infants and children with documented symptomatic UTI in several clinical areas in the Royal Hospital for Sick Children (RHSC) Glasgow. A total of 1,826 patient episodes were analysed, 1,592 of which were first referrals over a 29 month study period. The analysis highlighted a definite variation in a number of aspects of the management of UTI within the clinical services reviewed. In a significant number of in-patients, the diagnosis was made both in the absence of microbiological confirmation, and following the use of inappropriate urine specimens. An under-use of urine microscopy in the in-patient services was documented as well as an inappropriate use in out-patients, since the result in this latter group did not seem to influence the need for urine culture. Urinary tract imaging protocols varied widely between in-patient services and among in patient and out-patient services with a continued over-reliance on urinary tract ultrasound and an under-use of more sensitive radio-isotopic techniques. These data reveal a continuing lack of uniformity in management practices in children with UTI and demonstrate the need for a wider application of practice guidelines. PMID- 7490197 TI - An audit of medical paediatric out-patient appointments and letters. AB - All medical out-patient appointments of a firm of three consultants at the Royal Hospital for Sick Children were reviewed over a four week period. Of 758 appointments initially offered, 490 (64%) patients attended; 19% of patients cancelled and 17% did not attend without cancelling. Delay from referral to attendance ranged from 0 to 9 weeks. The characteristics of those patients experiencing prolonged registrar-only follow-up are described. The content of subsequent letters to family practitioners was usually adequate compared to accepted standards. Delays in dictation of letters were usually short in comparison to delays in typing. Such delays and the intervals between consultant review of out-patients may prove costly to providers. Potential improvement strategies are discussed. PMID- 7490198 TI - An audit of the accuracy of upper gastrointestinal diagnoses in Scottish Morbidity Record 1 data in Tayside. AB - INTRODUCTION: Scottish Morbidity Record (SMR1) data are coded by trained clerical staff from case records and discharge summaries. They form the basis of many strategic NHS decisions. Their accuracy for upper gastrointestinal (UGI) diagnosis is unknown and the study was undertaken to assess this accuracy in Tayside. METHOD: Patients who fulfilled the following criteria were identified using a record-linkage pharmacoepidemiological database, and their case records retrieved: over 50 years of age, had encashed at least one prescription for a non steroidal anti-inflammatory drug at a Tayside pharmacy and who had SMR1 records containing one or more symptom/diagnosis codes between January 1989 and December 1991. Medically qualified staff were trained to examine case records and to code UGI diagnoses. They searched the case records for every UGI SMR1 entry for these patients from 1980-1992 and produced re-coded diagnoses (RCD) for each hospital event (admission and discharge), using all the data available in the case records. They also abstracted data on the clinical presentation, investigations and management of patients. Each event was then examined by a single medically qualified researcher who compared the original SMR1 codes with the RCDs. RESULTS: 2,101 patients had a total of 3,764 events in 1989-1991. 317 events were either day case procedures or elective surgery or the case records were not found. They were therefore excluded. Of the remainder, the SMR1 and RCD codes were judged equivalent in 1,608 events (46.6%). However, 1,005 SMR1 events (29.2%) contained a symptom code but no diagnosis code and the remaining 834 (24.2%) were judged suboptimal for other reasons. Of those with a symptom code only, 406 could not be improved upon and were transformed into RCD symptom codes only, 435 were assigned symptom and diagnostic RCDs and 164 were assigned diagnostic RCDs only. In the other 834 events, 279 had one or more diagnoses missing, 425 had one or more diagnoses inaccurate, 23 had both missing and inaccurate diagnoses and 107 were not UGI. Thus 1,433 (41.6%) of UGI SMR1 events could be more accurately coded. Examination of investigation data revealed that coding inaccuracy was not due to diagnostic procedures being carried out after admission. CONCLUSION: UGI SMR1 data were satisfactory in about half of all events. In about a quarter there were symptom codes but no satisfactory diagnosis codes, whilst in another quarter the data were inaccurate. These findings have implications for health care activities and research that use these data. PMID- 7490199 TI - The epidemiology of leisure accidents in Scotland. PMID- 7490200 TI - Infertility-epidemiology, aetiology and effective management. AB - Current epidemiological evidence suggests that 15% of couples will experience infertility and in half this number the problem will remain unresolved. Background prevalence rates now appear to be reasonably stable, but there is evidence of an increase in the rate of referrals for medical help. Rates of secondary infertility are higher in the population than represented by clinic referrals. The distribution of the main diagnostic groups is as follows: Male 25%; Ovulation 25%; Tubal 20%; Unexplained 25%; and Endometriosis 5%. Effective management in the male includes donor insemination and assisted reproduction (including intra-cytoplasmic sperm injection). Drugs are ineffective for idiopathic oligozoospermia, while the role of varicocele ligation is uncertain, but marginal at best. Ovulation problems can be treated with a high degree of success, except in cases of clomiphene resistant polycystic ovarian disease. However there are continuing concerns over multiple pregnancy rates and future research is needed to clarify the additional risks, if any, of ovarian carcinoma. Surgical correction of tubal abnormalities should be left to specialised units, where audit indicates that the results can match those achieved by assisted reproduction. There is no evidence that the medical (drug) management of endometriosis improves fertility, although surgery for moderate and severe disease can still be considered. However the results should be compared with those achieved by assisted reproduction. Superovulation and intra-uterine insemination appears to be an effective treatment for certain cases of mild male factor infertility, mild endometriosis and unexplained infertility and can be considered, where the duration of infertility is more than four years. A pregnancy rate of around 10% per cycle can be anticipated, but there is a significant risk of multiple pregnancy. The evidence is that IVF (and GIFT) has achieved significantly improved results in recent years, with fecundity rates of 20% per cycle being a reasonable target in women under 40 years. PMID- 7490202 TI - The National Panel: past, present and future. PMID- 7490201 TI - The dietary habits of 11-12 year-old children in two Tayside secondary schools- comparison with the targets set by the Scottish Diet Report. AB - The diets of over 300 children in their first year at two Tayside secondary schools were assessed using a simplified food frequency questionnaire. Results were compared with the recent targets (Scottish Diet Report 1993) set for children for the year 2005. It was estimated that only six (< 2%) of the 331 children achieved all of the first five targets, namely 3-4 portions of vegetables and fruit per day; whole grain or granary bread or cereals twice daily; semi-skimmed milk; fish twice weekly; and meat products less than twice weekly. In addition, less than 5% achieved a Healthy Food Choice score (devised by the authors) commensurate with a healthy diet containing daily fruit and vegetables, semi-skimmed milk, wholemeal or similar bread or cereals, and fish at least once a week. PMID- 7490203 TI - Intra-oral cancers in Scotland. PMID- 7490204 TI - Scottish neonatal intensive care units; a study of staff and parental attitudes. AB - The study takes a three stage approach to review Scottish unit policies, staff views and parents experiences of visiting and involvement with their low birthweight infant (1,750 gm or less) in a sample of Scottish neonatal intensive care units (NICUs). The study indicated that while most units had 'liberal' views regarding parental and family visiting some staff views were not in line with their unit policy and retained reservations about family visiting. Many parents reported visiting daily despite considerable 'social' and real costs. Unit differences emerged regarding the degree of parental involvement as measured by tasks carried out. PMID- 7490205 TI - 'Video, health education and the General Practitioner Contract'. AB - The 1990 General Practitioner Contract and the 1993 Contract Revision emphasised the role of health education/health promotion within primary care. In this paper, the authors suggest that a highly cost-effective approach is necessary given the premium on time available for general practitioners and members of the primary health care team to undertake such work. The views of general practitioners and the primary health care team in a random sample of general practices within all 15 Health Board areas in Scotland were sought between 1991 and 1993 regarding current experience and future potential in using video-based health education materials, with particular emphasis on the content, structure and parameters appropriate to running effective health education/health promotion programmes. This study has indicated the potential demand for high quality, appropriately targeted video-based materials for use by primary health care workers within a range of practice, community and home settings. Central to this is the need for a better system of information and dissemination of materials and a firmer evaluation framework for health education/health promotion programmes within primary care. The progress of several policy, management and research issues which arose from the findings of the study have been raised by the authors. These include implications for the NHS such as infrastructure resourcing and priority setting, programme development, information and dissemination, research and evaluation, and mechanism(s) whereby general practitioners can be better involved in this process. PMID- 7490206 TI - BMS-192548, a tetracyclic binding inhibitor of neuropeptide Y receptors, from Aspergillus niger WB2346. I. Taxonomy, fermentation, isolation and biological activity. AB - During the screening of microbial fermentation extracts for their ability to inhibit the binding of 125I-peptid YY (PYY) to the neuropeptide Y (NPY) receptor using the scintillation proximity assay (SPA), BMS-192548 was isolated from the extract of Aspergillus niger WB2346 by bioassay-guided fractionation. BMS-192548 showed the inhibitory activity against 125I-PYY binding to SK-N-MC and SMS-KAN cells, which express NPY1 and NPY2 receptors, respectively, with IC50 values of 24 microM in Y1 and 27 microM in Y2 receptor binding. BMS-192548 demonstrated weak cytotoxicity against murine tumor cell line M-109 with an IC50 value of 240 microM. PMID- 7490207 TI - BMS-192548, a tetracyclic binding inhibitor of neuropeptide Y receptors, from Aspergillus niger WB2346. II. Physico-chemical properties and structural characterization. AB - The structure of BMS-192548, a tetracyclic binding inhibitor of neuropeptide Y receptors, was established by spectroscopic methods. The compound has an unusual B-C-D ring beta-diketone moiety. PMID- 7490208 TI - WS75624 A and B, new endothelin converting enzyme inhibitors isolated from Saccharothrix sp. No. 75624. I. Taxonomy, fermentation, isolation, physico chemical properties and biological activities. AB - Novel endothelin converting enzyme (ECE) inhibitors, WS75624 A and B, have been isolated from the fermentation broth of Saccharothrix sp. No. 75624. These inhibitors were purified from an acetone extract of whole culture broth followed by HP-20 column chromatography, silica gel column chromatography and HPLC. WS75624 A and B showed highly potent ECE inhibitory activity, and both had IC50 values of 0.03 microgram/ml. WS75624 A and B also showed other metalloprotease (collagenase and neutral endopeptidase) inhibitory activity with IC50 values of 1 microgram/ml. Since large amount of WS75624 B was isolated, we tried in vivo evaluation using WS75624 B. WS75624 B inhibited big endothelin-induced pressor effect when administered to SD rat intravenously with big ET-1. PMID- 7490209 TI - WS75624 A and B, new endothelin converting enzyme inhibitors isolated from Saccharothrix sp. No. 75624. II. Structure elucidation of WS75624 A and B. AB - The structures of WS75624 A and B, novel endothelin converting enzyme inhibitors, were determined to be 1 and 2A, respectively, by a combination of chemical evidence and a series of 2D NMR spectral analyses. PMID- 7490211 TI - Isolation and structure determination of two novel phenazines from a Streptomyces with inhibitory activity against metallo-enzymes, including metallo-beta lactamase. AB - Two novel metabolites, SB 212021 and SB 212305, have been isolated from a Streptomyces and shown to have molecular formulae of C15H10N2O5 and C20H17N3O8S, respectively. The structures were deduced by a combination of NMR techniques and mass spectral fragmentation patterns and shown to be novel members of the phenazine group of antibiotics. In the absence of added zinc, both compounds had IC50's of 1-75 microM for the Bacteroides fragilis 262 CfiA and Xanthomonas maltophilia L-1 metallo-beta-lactamases. The compounds also inhibited ACE with IC50's of 55 and 68 microM, respectively. Mode of action studies illustrate that the compounds inhibit some metalloenzymes by chelation of the active site metal ion. They exhibit poor antibacterial activity. PMID- 7490210 TI - Cochlioquinone A, an inhibitor of diacylglycerol kinase. AB - The effects of cochlioquinone A, isolated from Drechslera sacchari, were studied in vitro and in vivo. This compound specifically inhibited diacylglycerol kinase activity with Ki = 3.1 microM. The kinetics revealed that cochlioquinone A inhibited diacylglycerol kinase in competition with ATP, and non-competitively with diacylglycerol. The compound inhibited neither protein kinase C, epidermal growth factor receptor-associated protein tyrosine kinase, nor phospholipase C. Cochlioquinone A reduced the concentration of phosphatidic acid in T cell lymphoma with a half maximal concentration of 3 microM, and simultaneously augmented the phosphorylation of 80 kDa protein, a known substrate of protein kinase C. The degree of the phosphorylation of 80 kDa protein in the presence of cochlioquinone A was similar to that in the presence of phorbol myristate acetate (0.1 microgram/ml). These results demonstrate that cochlioquinone A is a specific inhibitor of diacylglycerol kinase, which regulates the activity of protein kinase C. PMID- 7490212 TI - Louisianins A, B, C and D: non-steroidal growth inhibitors of testosterone responsive SC 115 cells. I. Taxonomy, fermentation, isolation and biological characteristics. AB - In the course of screening for non-steroidal growth inhibitors of testosterone responsive Shionogi carcinoma 115 cells, louisianins A, B, C and D, were isolated from fermentation broth of Streptomyces sp. WK-4028. Louisianin A remarkably inhibited the growth of SC 115 cells in the presence of 10(-7) M testosterone at an IC50 value of 0.6 microgram/ml, whereas, no inhibition was observed on the other cell lines. Furthermore, inhibitory activity of testosterone 5 alpha reductase and antimicrobial activity have not been observed at a concentration of 50 micrograms/ml and 1,000 micrograms/ml, respectively. PMID- 7490213 TI - Louisianins A, B, C and D: non-steroidal growth inhibitors of testosterone responsive SC 115 cells. II. Physico-chemical properties and structural elucidation. AB - New non-steroidal growth inhibitors of testosterone-responsive SC 115 cells, louisianins A (MW: 189; C11H11NO2), B (MW: 191; C11H13NO2), C (MW: 173; C11H11NO) and D (MW: 173; C11H11NO) were isolated from the cultured broth of Streptomyces sp. WK-4028. Their structures were determined on the basis of spectroscopic data. The structure of louisianin A in particular was confirmed by X-ray crystallographic analysis. The four compounds commonly possess a unique pyrindine skeleton in the molecule. PMID- 7490214 TI - Bacillopeptins, new cyclic lipopeptide antibiotics from Bacillus subtilis FR-2. AB - Bacilopeptins, new iturin-group antifungal antibiotics, were isolated from the culture broth of Bacillus subtilis FR-2 obtained from the rhizosphere of garlic suffering from the basal rot caused by Fusarium oxysporum. Their structures were elucidated to be cyclic lipopeptides similar to bacillomycin L by NMR and mass spectral studies coupled with amino acid analysis. The absolute configuration of each amino acid residue was determined by chiral HPLC. PMID- 7490215 TI - Tetrodecamycin and dihydrotetrodecamycin, new antimicrobial antibiotics against Pasteurella piscicida produced by Streptomyces nashvillensis MJ885-mF8. I. Taxonomy, fermentation, isolation, characterization and biological activities. AB - The novel antimicrobial antibiotic against Pasteurella piscicida, tetrodecamycin (1) and weakly active dihydrotetrodecamycin (2) were isolated from the fermentation broth of Streptomyces nashvillensis MJ885-mF8. They were purified by adsorption on Diaion HP-20, silica gel column chromatography and crystallization. The MICs of 1 were 6.25 approximately 12.5 micrograms/liter and 1.56 approximately 6.25 micrograms/ml against Gram-positive bacteria including methicillin-resistant Staphylococcus aureus (MRSA) and 12 strains of P. piscicida, respectively. PMID- 7490216 TI - Tetrodecamycin and dihydrotetrodecamycin, new antimicrobial antibiotics against Pasteurella piscicida produced by Streptomyces nashvillensis MJ885-mF8. II. Structure determination. AB - Novel antimicrobial antibiotics against Pasteurella piscicida, tetrodecamycin (1) and weakly active dihydrotetrodecamycin (2) were isolated from a culture broth of Streptomyces nashvillensis MJ885-mF8. The planar structure of 1 was determined to be 2-acyl-4-ylidene tetronic acid alkyl ether containing decaline ring by various NMR spectral data of 1 and its acetyl derivative (3). The structure of 2 was elucidated by comparison with the spectral data of 1 and confirmed by catalytic reduction of 1 into 2. The X-ray crystallography of 2 showed the relative stereochemistry. Their absolute configurations were determined by using modified Mosher's method. PMID- 7490217 TI - Structure of new cyclic depsipeptides, Bu-2841-08 and -10. AB - The structures of new antibiotics, Bu-2841-08 and -10, have been determined. They are cyclic depsipeptides and the sequence of amino acid residues was established by mass spectral analysis of the hydrolyzed linear peptide and NMR spectral analysis of the parent cyclic peptides. PMID- 7490218 TI - Phosphonothrixin, a novel herbicidal antibiotic produced by Saccharothrix sp. ST 888. I. Taxonomy, fermentation, isolation and biological properties. AB - A novel herbicidal compound, phosphonothrixin, was found in the fermentation broth of Saccharothrix sp. ST-888 cultured on a vegetable juice medium. The compound exhibiting acidic and hydrophilic properties was obtained when the fermentation broth of ST-888 was subjected to ion exchange chromatography, gel filtration chromatography and ion-pair chromatography. Phosphonothrixin significantly inhibited germination of gramineous and broadleaf weeds. Foliar application of this antibiotic gave rise to chlorosis in all of the plants tested. PMID- 7490219 TI - Phosphonothrixin, a novel herbicidal antibiotic produced by Saccharothrix sp. ST 888. II. Structure determination. AB - Phosphonothrixin (1) is a novel herbicidal agent produced by Saccharothrix sp. ST 888. This unique compound possessing a C-P bond and an isoprene unit was determined to be 2-hydroxy-2-hydroxymethyl-3-oxobutyl-phosphonic acid. Unusually rapid proton-deuterium (H-D) exchange of the methyl ketone unit of this antibiotic in D2O solution was observed. Phosphonothrixin can be classified as a new type of natural product herbicide having a C-P bond in the molecule. PMID- 7490220 TI - Total synthesis of (+/-)-phosphonothrixin, a novel herbicidal antibiotic containing C-P bond. AB - Phosphonothrixin (1) is a novel herbicidal agent produced by Saccharothrix sp. ST 888, with a unique structure possessing a C-P bond and an isoprene unit. The total synthesis of this antibiotic was accomplished from methyl (bromomethyl)acrylate in six steps. PMID- 7490221 TI - Screening for apoptosis inducers in microbial products and induction of apoptosis by cytostatin. AB - We have employed the DNA-methylgreen binding assay as a primary screening method for identifying apoptosis inducers in microbial products. Capsimycin, toyocamycin and cytostatin affect the binding of methylgreen to DNA of FS3 cells in this test system. The effect of cytostatin on apoptosis induction was confirmed by means of the ELISA system. PMID- 7490223 TI - Azicemicins A and B, new antimicrobial agents produced by Amycolatopsis. II. Structure determination. AB - A new structural class of antibiotics, azicemicins A (1) and B (2) were isolated from the culture broth of Amycolatopsis sp. MJ126-NF4. Their structures were elucidated from their physico-chemical properties, various NMR experiments and chemical transformations and were shown to be 3-(1-methyl-2-aziridinyl)- and 3-(2 aziridinyl)-3,4-dihydro-3,7,8,10,12b-pentahydroxy-9,12-dimeth oxy-benz [a]anthracene-1,6(2H,5H)-dione, respectively. PMID- 7490222 TI - In vivo evaluation of three acid-stable azalide compounds, L-701,677, L-708,299 and L-708,365 compared to erythromycin, azithromycin and clarithromycin. AB - L-701,677, L-708,299 and L-708,365 are novel azalide derivatives of erythromycin that exhibit improved acid stability over erythromycin, azithromycin and clarithromycin. The half-life in aqueous solution at pH = 2.1 of these compounds ranged from 0.3 hour for erythromycin to 16.2 hours for L-708,299. The rank order of half-life in acid solution from most to least stable was L-708,299 > L-701,677 > L-708,365 > azithromycin = clarithromycin > erythromycin. In a disseminated Streptococcus pyogenes mouse infection model, azithromycin and L-708,365 were slightly more efficacious than clarithromycin, L-701,677 and L-708,299; all 5 compounds being more active than erythromycin. In a Klebsiella pneumoniae pulmonary challenge mouse model, azithromycin, L-701,677, L-708,299 and L-708,365 were all equal in efficacy and at least four-fold more active than clarithromycin and erythromycin. Clarithromycin, L-708,365 and interestingly erythromycin, showed greater bacterial clearance than azithromycin, L-701,677 and L-708,299 in a localized infection model that measured clearance of Staphylococcus aureus from mouse thigh tissues. Our results indicate that L-701,677, L-708,299 and L-708,365 exhibit improved acid stability and were at least equally efficacious as presently marketed macrolide/azalide antibiotics. PMID- 7490224 TI - New anthracycline antibiotics 10-epi-oxaunomycin and 10-epi-11-deoxyoxaunomycin. AB - Two new anthracycline antibiotics, 10-epi-oxaunomycin and 10-epi-11 deoxyoxaunomycin, were photochemically obtained from anthracycline metabolites D788-1 (10-carboxy-13-deoxocarminomy-cin) and D788-3 (10-carboxy-11-deoxy-13 deoxocarminomycin) and were examined for their growth inhibitory activities on cultured L1210 leukemic cells. Effects of the S configuration of C-10 and a hydroxyl group at C-11 on the bioactivity are discussed in comparison with oxaunomycin and 11-deoxyoxaunomycin. PMID- 7490225 TI - Structure-activity relationship of pamamycins: effects of alkyl substituents. AB - Nine new pamamycin homologues were isolated from the culture broth of Streptomyces alboniger IFO 12738 using a combination of ODS and NH2 HPLCs, and their structures determined by GC-MS. The structural differences in these homologues are in the numbers and positions of methyl and ethyl groups. The aerial mycelium-inducing and growth-inhibitory activities in S. alboniger of these homologues and their antibiotic activity against Bacillus subtilis were examined. The effects of the alkyl substituents on these activities are discussed. PMID- 7490226 TI - Phevalin, a new calpain inhibitor, from a Streptomyces sp. PMID- 7490227 TI - MR304A, a new melanin synthesis inhibitor produced by Trichoderma harzianum. PMID- 7490228 TI - SCH57404, an antifungal agent possessing the rare sodaricin skeleton and a tricyclic sugar moiety. PMID- 7490229 TI - TMC-34, a new macrolide antifungal antibiotic. PMID- 7490230 TI - Preparation of optically active epiderstatin and its stereoisomers--epiderstatin is not a real inhibitor of the mitogenic activity induced by epidermal growth factor. PMID- 7490231 TI - Thiazoline analogues of epiderstatin, new inhibitors of cell cycle of tsFT-210 cells. PMID- 7490232 TI - Selective cytotoxicity of mycophenolic acid against transformed cells. PMID- 7490233 TI - Synthesis and antiproliferative effects of a 4'-morpholino-9-methyl anthracycline. PMID- 7490234 TI - Phomactin E, F, and G: new phomactin-group PAF antagonists from a marine fungus Phoma sp. PMID- 7490235 TI - Nucleotide sequence of fortimicin KL1 methyltransferase gene isolated from Micromonospora olivasterospora, and comparison of its deduced amino acid sequence with those of methyltransferases involved in the biosynthesis of bialaphos and fosfomycin. PMID- 7490236 TI - Surgical management of primary soft tissue sarcoma. AB - Few areas in oncology more clearly illustrate the need for multidisciplinary management than the soft tissue sarcoma problem. Using this approach has led to limb preservation with survival results at least equivalent to those for amputations. This achievement has dramatically improved the quality of life and overall clinical outcome for patients with this disease. In the future, improved understanding of the biology of this disease may broaden our therapeutic options. Meanwhile, a detailed understanding of the surgical possibilities will enhance the multimodality care that can be offered by the teams of clinicians managing patients suffering from this disease. PMID- 7490237 TI - Surgical management of primary bone sarcomas. AB - Effective chemotherapy regimens have changed the nature and scope of surgical management of primary sarcomas of bone. Limb-salvage surgical procedures are performed in the majority of patients without a significantly increased risk of local tumor relapse. Advances in technology have expanded the options available for skeletal reconstruction. End-result analyses have helped refine the application of this technology to maximize function. Further advances are necessary to maintain function in the long term. PMID- 7490238 TI - Radiation as a therapeutic modality in sarcomas of the soft tissue. AB - Management of the primary lesion in patients with soft tissue sarcomas is combined modality in the majority of instances. The modalities are surgery and irradiation. Radical surgery as currently practiced in major oncology centers does not achieve local control in an important proportion of patients. PMID- 7490239 TI - Brachytherapy in sarcomas. AB - Historically, amputation was the standard local treatment for soft tissue sarcomas of the extremity. Although this procedure provided excellent local control, the adverse impact of this procedure on quality of life was obvious. In an effort to maximize the functional outcome of sarcoma therapy, various groups initiated studies aimed at doing less radical operations in conjunction with radiation therapy. PMID- 7490240 TI - Results of single-agent and combination chemotherapy for advanced soft tissue sarcomas. Implications for decision making in the clinic. AB - Cytotoxic chemotherapy remains the principal modality of treating advanced or unresectable sarcomas of soft tissue. Certain drugs, such as doxorubicin and ifosfamide, are able to induce clinically useful antitumor effects as single agents, and combination chemotherapy regimens have been tested based upon these results. This article summarizes the results of single agent and combination chemotherapy trials for soft tissue sarcomas. PMID- 7490241 TI - Systemic adjuvant chemotherapy for soft tissue sarcomas. AB - Adjuvant chemotherapy for soft tissue sarcoma remains investigational. Continued study of multidisciplinary treatment programs in high-grade sarcomas in formal clinical trials remains a priority. The National Cancer Institute is currently conducting a randomized trial of postoperative adjuvant doxorubicin and ifosfamide as compared with observation in high-grade sarcoma. The completion and outcome of this trial will contribute to this ongoing debate. Finally, a better understanding of the biology of soft tissue sarcoma that determines subsequent clinical course may serve as a foundation for further investigation. PMID- 7490242 TI - Adjuvant chemotherapy for soft tissue sarcomas. AB - Despite considerable interest in this subject and the completion of a number of randomized trials, the use of adjuvant chemotherapy after definitive local treatment for patients with soft tissue sarcomas remains problematic. This article reviews randomized clinical trials completed and published to date, and explores new strategies for future studies. PMID- 7490243 TI - Preoperative therapy for soft tissue sarcoma. AB - Soft tissue sarcomas appear to be an ideal tumor type for delivering preoperative therapy. The rationale for preoperative therapy is that it is delivered to undisturbed tissue planes with well-oxygenated tissue. This is of great benefit for radiation therapy, because with new computed tomography scan treatment planning it is possible to completely delineate the tumor without surgical clips or postoperative hematoma (or both) obscuring the tumor margin. PMID- 7490245 TI - Osteosarcoma. Specimen management following primary chemotherapy. AB - The use of preoperative (i.e., primary) chemotherapy has been shown to be of direct and significant benefit to the osteosarcoma patient. There is virtually an immediate palliation of symptoms. Increasing numbers of patients with osteosarcoma involving bones of the appendicular skeleton are eligible for limb salvage procedures secondary to the local effects of chemotherapy that result in a decrease in size and compaction (i.e., down-staging) of the tumors. PMID- 7490244 TI - Chemotherapy in osteosarcoma. Basis for application and antagonism to implementation; early controversies surrounding its implementation. AB - Chemotherapy in osteosarcoma has had a major impact in attaining metastasis-free survival, and the cure rates with the use of modern regimens range from 65% to 75%. Preoperative chemotherapy has facilitated surgical resection of the primary tumor, rendering increased numbers of candidates for limb salvage. The effects on the primary tumor may be used as a predictive factor in determining outcome. PMID- 7490246 TI - Surgical treatment for pulmonary metastases from sarcoma. AB - Pulmonary metastases from soft tissue sarcomas frequently metastasize to the lungs and may be resected safely. In approximately 30% of patients, long-term survival is achieved. Prognostic indicators may define clinically the biology of the metastasis. Use of various prognostic indicators may assist the clinician in selecting the best treatment plan for these patients. PMID- 7490247 TI - Multidrug drug resistance in pediatric sarcomas. AB - PURPOSE: Although P-glycoprotein overexpression is an important cause of multidrug resistance in vitro, whether this resistance mechanism is equally applicable to the clinic has still to be fully established. This review has examined the immunohistochemical and molecular biologic tools used to evaluate soft tissue sarcomas of children and adults, in order to determine whether P glycoprotein can limit the efficacy of chemotherapy. DESIGN: Because soft tissue sarcomas are successfully treated by cytotoxic substrates of P-glycoprotein in many, but not all children, these tumors may be useful models for determining whether this drug efflux transporter is a clinically relevant cause of chemoresistance. RESULTS: Certain studies of acute myelogenous leukemia, lymphoma, and myeloma in adults and rhabdomyosarcoma, neuroblastoma, and acute lymphoblastic leukemia in children have provided the best current evidence for a strong corelation between the expression of P-glycoprotein and outcome of chemotherapy. Based on these observations, several clinical trials have been initiated to determine whether pharmacologic chemosensitization improves chemotherapy responses and cure rates in P-glycoprotein-expressing malignancies. Thus, early identification of lower levels of P-glycoprotein in tumors that still might respond to chemosensitizer-modulated chemotherapy may be highly pertinent to conducting more informative multidrug resistance reversal trials. CONCLUSION: The question of whether the multidrug resistance P-glycoprotein is a clinically relevant cause of chemoresistance may ultimately be answered by the successful prevention of chemotherapy failure by chemosensitizers that specifically reverse this drug efflux mechanism. PMID- 7490248 TI - New drugs for the treatment of sarcomas. AB - Despite the fact that many new active drugs recently became available for phase II studies of the different sarcoma types, only soft tissue sarcoma has been included in these phase II testing programs. The groups active in the treatment of osteosarcomas and Ewing's sarcomas should be encouraged to investigate the new active drugs in the relapsing patients. For soft tissue sarcoma, many new drugs have been tested by different groups. The results to date have been disappointing, because only docetaxel has been identified as a new active drug in one phase II study. Hints of activity have also been observed for edatrexate and topotecan. Further studies with these three drugs are necessary to identify their proper role. PMID- 7490250 TI - Exploring the nucleotide-binding site in proteins by affinity labeling and site directed mutagenesis. AB - Affinity labeling with nucleoside polyphosphopyridoxals, especially those with 3 or 4 phosphate groups, was effective for identifying the lysyl residue(s) located at or near the binding site for ATP, GTP, UDP-Glc, or ADP-Glc in various proteins. Furthermore, kinetic analysis of the mutant enzymes, in which the labeled lysyl residue was replaced with another amino acid by site-directed mutagenesis, provided evidence of its functional role. Affinity labeling of the mutant enzymes was useful for further identification of the hidden lysyl residue, which is unreactive in the wild-type enzyme but catalytically important. Comparison of the results of affinity labeling with different substrate analogues provided the information on the location of the labeled lysyl residue around the bound substrate. The affinity labeling reflected structural features of proteins, including their conformational flexibility. PMID- 7490249 TI - Biologic therapy for osteosarcoma using liposome-encapsulated muramyl tripeptide. AB - The successful treatment of metastases will have to include modalities that can overcome the obstacles presented by the heterogeneous nature of malignant neoplasms and the continuous evolution of variant cells. Macrophages activated to become tumoricidal by interaction with L-MTP-PE may be able to accomplish this. Osteosarcoma appears to be an ideal disease in which to employ L-MTP-PE as an additional adjuvant to present chemotherapy regimens. The lung is the most frequent site of metastases, and pulmonary micrometastases are considered to be present in the majority of patients at diagnosis. Approximately 40% of patients with osteosarcoma develop pulmonary metastases despite the administration of adjuvant chemotherapy. The 2-year disease-free interval has not improved over the past 10 years, despite multiple changes in adjuvant regimens. These data argue that there is a subpopulation of patients who harbor tumor cells that are relatively resistant to all chemotherapy. Unfortunately, this group of patients cannot be identified at the time of initial diagnosis. This necessitates the incorporation of new forms of therapy into the adjuvant chemotherapy protocols for all patients in the hope of eradicating the resistant cells harbored in the 40%. Based on the data summarized previously, L-MTP-PE may improve the clinical outcome of patients with osteosarcoma by activating pulmonary macrophages to destroy residual tumor cells that are not eliminated by chemotherapy. Monocytes from osteosarcoma patients can be rendered cytotoxic to tumor cells by in-vitro incubation with L-MTP-PE and following the intravenous administration of this agent. L-MTP-PE can be given safely to both adults and children with minimal side effects. The whole-body distribution of 99mTc-labeled liposomes containing MTP-PE confirms that the agent is taken up by the lungs. Biologic activity in osteosarcoma patients is revealed by the elevations in plasma levels of several cytokines plus stimulation of monocyte-mediated cytotoxicity following L-MTP-PE infusion and by histologic changes in the pulmonary lesions. Ifosfamide therapy given in combination with L-MTP-PE does not suppress this immune response, as judged by both plasma cytokine levels and tumor histology. Finally, L-MTP-PE has been shown to be effective as a single agent against relapsed osteosarcoma. It is unlikely that the addition of other chemotherapeutic agents to the adjuvant chemotherapy protocols will alter the 65% to 70% 2-year disease-free survival rate associated with osteosarcoma. The preceding data indicate that L-MTP-PE is an active agent against this disease and deserves further investigation. Therefore, the inclusion of L-MTP-PE with chemotherapy is a reasonable alternative to consider to improve the response rate of this disease.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 7490251 TI - Novel sensing system for glucose based on the complex formation between phenylborate and fluorescent diol compounds. AB - From the standpoints of long-term stability and simple handling, a glucose sensor composed of totally synthetic sensing moieties should be of great advantage. This study was performed to demonstrate the feasibility of a novel glucose sensing system in an aqueous milieu based on a change in the fluorescence on competitive binding between a fluorescent diol compound and glucose toward a phenylborate compound. 6,7-Dihydroxy-4-methyl-coumarin (DHMC) was selected as a fluorophore, and 3-propionamidophenylboronic acid (PAPBA) as a phenylborate compound. The relationship between the glucose concentration and fluorescence intensity was linear in the range of 0 to 500 mg/dl of glucose. The availability of various derivatives of DHMC and phenylborate compounds allows the incorporation of these functionalities on the tip of an optical fiber to construct an opto-sensing system for glucose. PMID- 7490252 TI - A putative metal-binding site in the beta subunit of rat mitochondrial processing peptidase is essential for its catalytic activity. AB - Mitochondrial processing peptidase (MPP) consists of alpha- and beta-subunits (alpha-MPP and beta-MPP). beta-MPP has a putative metal-binding sequence (HXXEH). To determine whether the sequence of beta-MPP is essential for the enzymatic activity, we individually mutated the histidines and glutamic acid to arginines and glutamine, respectively. The wild-type and mutated beta-MPPs were co expressed with alpha-MPP in Escherichia coli. All three mutants had completely lost the activity, whereas the lost activity was recovered on the addition of wild-type beta-MPP. The activity of the wild-type enzyme was reduced by the mutant beta-MPPs. We conclude from these observations that the HXXEH region is involved in the formation of the active site and that beta-MPP is the catalytic subunit of MPP. PMID- 7490253 TI - Morphology of a cytochrome c-adsorbed stearic acid monolayer on Brewster angle microscopy. AB - The morphologies of stearic acid and cytochrome c (cyt.c)-adsorbed stearic acid monolayers were investigated by Brewster angle microscopy (BAM) with various molecular areas of stearic acid. With an area of more than 0.38 nm2/molecule, many blight island domains and some bright circles were observed in the BAM image of the stearic acid monolayer. The blight site part became to occupy all the surface with compression, and then became more closely packed with an area of 0.22 nm2/molecule. On the other hand, a different BAM image was obtained for the cyt.c-adsorbed stearic acid monolayer, as follows: (i) a striped pattern was only observed in the presence of cyt.c; (ii) the number of bright circles in the presence of cyt.c was less than that in its absence. Furthermore, when a uniform BAM image was observed for the stearic acid monolayer with cyt.c, the intensity of the absorbance at 409 nm of cyt.c was the highest. By calculating the amount of cyt.c adsorbed on a stearic acid monolayer from the absorbance value, it was shown that cyt.c was most closely packed when an uniform BAM image was observed. These results suggest that the use of BAM and visible absorption spectroscopy together is useful for studying the morphology of a monolayer. PMID- 7490254 TI - The role of tyrosine phosphorylation and Ca2+ accumulation in Fc gamma-receptor mediated phagocytosis of human neutrophils. AB - When stimulated with an insoluble immune complex consisting of tetanus toxoid and human anti-tetanus toxoid IgG antibody, human neutrophils recognized the immune complex via Fc receptors (Fc gamma RII and Fc gamma RIIIB) on their plasma membranes, resulting in Ca2+ accumulation, and finally phagocytosed the immune complex. Ca2+ depletion experiments suggest that Ca2+ accumulation is essential for the maximal phagocytic activity. A tyrosine kinase inhibitor, genistein, inhibited the phagocytosis of immune complex. Genistein also inhibited both the later prolonged Ca2+ accumulation below 100 microM, and the initial rapid Ca2+ accumulation above 100 microM. Stimulation of human neutrophils by immune complexes through Fc gamma Rs caused tyrosine phosphorylation of multiple neutrophil proteins within 15 s, proceeding to an increase in the intracellular Ca2+ concentration. A similar pattern of tyrosine phosphorylation was observed when Fc gamma RII or Fc gamma RIIIB was aggregated by an anti-Fc gamma R monoclonal antibody and anti-mouse Ig. These results suggest that tyrosine phosphorylation mediates, at least in part, the later responses (Ca2+ accumulation and phagocytosis) of the cells and is an important early step in signaling via Fc gamma Rs. PMID- 7490255 TI - Reaction of 20S proteasome: shift of SDS-dependent activation profile by divalent cations. AB - The multicatalytic endopeptidase complex (20S proteasome) is a latent high molecular-mass multisubunit proteinase. In many investigations, SDS has been used as a proteasome activator at some fixed concentration that was apparently optimal. This study examined the effects of various divalent cations on the SDS dependent peptidase and casein degradation activities of 20S proteasome purified from Xenopus laevis oocytes at a series of SDS concentrations and the correlation between these effects and the critical micelle concentration (CMC) of SDS. Surprisingly, it was found that divalent cations such as Mg2+ markedly shifted the SDS-dependent activation profiles to a lower concentration range. Ca2+, Mn2+, Co2+, and Zn2+ also markedly reduced the optimum SDS concentration in the Suc-Leu Leu-Val-Tyr-MCA hydrolysis reaction: for example, 5 mM Co2+ reduced the optimum SDS concentration from 0.065 to 0.005%. However, in all cases examined the optimum concentrations were below the CMC. Cu2+, Hg2+, and Cd2+ strongly inhibited the SDS-dependent maximum activity without remarkably shifting the optimum SDS concentration. No correlation between the shift and the inhibition was recognized. Most interestingly, remarkable activation of casein degradation by SDS was observed only by addition of the divalent cations Mg2+, Ca2+, and Mn2+. These cations might be essential for casein degradation. The activation and inactivation ranges of SDS concentration varied with the species of substrate.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7490256 TI - Induction of heat shock protein 70 and nucleolin and their intracellular distribution during early stage of liver regeneration. AB - Our previous work demonstrated a simultaneous induction of Hsp70 and nucleolin mRNA during the prereplicative stage of hepatocytes after partial hepatectomy [Ohmori, H. et al. (1990) Exp. Cell Res. 189, 227-232]. In the present study, changes of intracellular localization of these proteins were examined. Nucleolin, which mainly localized in the nucleus, increased with time and accumulated in the nucleolus around 12-18 h after partial hepatectomy. Hsp70 protein also increased slightly around 6-12 h after the operation. In accordance with this increase, immunohistochemical staining revealed that almost all nuclei of hepatocytes became Hsp70-positive, although Hsp70 was seen to be dispersed throughout the nucleoplasm and nucleolus at all times examined. Next, we isolated a cDNA clone of ribosomal protein (S-17) and examined its behavior. Induction of S-17 mRNA was observed to be essentially similar to that of nucleolin mRNA in regenerating rat liver, although inductions of histone H2A and H4 occurred at a later time, that is, in parallel with DNA synthesis. Furthermore, we observed a simultaneous induction of Hsp70 and nucleolin mRNA by serum-stimulation after serum-depletion in HeLa and IAR-20 (rat) culture cells. These results suggest that the induction of Hsp70, in addition to nucleolin, was not fortuitous but may be involved in the early events of liver regeneration. PMID- 7490257 TI - Mitochondrial alcohol dehydrogenase from ethanol-grown Euglena gracilis. AB - The inducing effects of ethanol on alcohol dehydrogenase and the key enzymes of the glyoxylate cycle, isocitrate lyase and malate synthase, in Euglena cells were investigated. Ethanol as the sole carbon source resulted in increases in alcohol dehydrogenase and the two glyoxylate cycle enzymes. The experimental results indicated that ethanol is assimilated by alcohol dehydrogenase and the glyoxylate cycle in Euglena. Mitochondria from aerobically grown Euglena contain a unique type of alcohol dehydrogenase that accounts for their ability to respire with ethanol as a substrate. This alcohol dehydrogenase was purified to homogeneity from ethanol-grown Euglena gracilis. The mitochondrial alcohol dehydrogenase was NAD(+)-specific but not NADP(+)-specific. Ethanol was the most active substrate, but the enzyme was also active towards 1-butanol, 1-heptanol, cinnamyl alcohol, and myristyl alcohol. These results indicated that mitochondrial alcohol dehydrogenase participated in alcohol metabolism in Euglena gracilis. PMID- 7490258 TI - Characterization of vitelline membrane outer layer protein I, VMO-I: amino acid sequence and structural stability. AB - Vitelline membrane outer layer protein I (VMO-I) tightly bound to ovomucin fibrils of hen's egg yolk membrane was characterized in terms of its amino acid sequence and structural stability. The deduced sequence of VMO-I using the conventional sequencing method is: RTREYTSVITVPNGGHWGKWGIRQFCHSGYANGFALKVEPSQFGRDDTALNGIRLRCLD- GSVIESLVGKWGTWTSFLVCPTGYLVSFSLRSEKSQGGGDDTAANNIQFRCSDEAVLVGD- DLSWGRFGPWSKRCKICGLQTKVESPQGLRDDTALNNVRFFCCK. Thus, VMO-I is composed of 163 amino acid residues with a calculated molecular weight of 17,979. The sequence confirms the cDNA sequence of VMO-I we recently determined and does not show any significant similarity to proteins compiled in the NBRF database. Two of the four disulfide bonds found in VMO-I were estimated to lie between Cys26 and Cys57 and between Cys79 and Cys110. The sequence analyses show that VMO-I contains three 53 residue internal repeats that contain distinctive regions of turns flanked by beta-sheets consistent with the recent finding that the molecule contains a new beta-fold motif, the beta-prism. The molecular characteristics of VMO-I in solution were examined by CD spectroscopy in the far and near ultraviolet regions, NMR spectroscopy, and high sensitive differential scanning calorimetry (DSC). CD spectra in the far UV region at room temperature were similar to that assigned to a random coil, while in the near UV region, small positive peaks were observed. The ellipticity in both regions decreased on raising the temperature. Proton NMR experiments showed the native structure unfolds to unordered conformations at 70 degrees C.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7490259 TI - Determination of soluble nAChR-binding activity of alpha-neurotoxins by an innovative precipitation with DEAE-Sephacel. AB - A simple and convenient method for determining the binding activity of alpha neurotoxins toward soluble nicotinic acetylcholine receptor (snAChR) by precipitation with DEAE-Sephacel was established. The determination was carried out by incubation of 125I-neurotoxin with snAChR, followed by precipitation with DEAE-Sephacel. The DEAE-Sephacel particles bind negatively charged snAChR with high affinity and simultaneously precipitate the 125I-neurotoxin bound to the receptors. After centrifugation, the free 125I-neurotoxin in the supernatant was counted, and the amounts of neurotoxins bound to snAChR could be determined. Two alpha-neurotoxins, cobrotoxin and alpha-bungarotoxin, were employed to verify the feasibility of this determination. The different binding characteristics of cobrotoxin and alpha-bungarotoxin to snAChR could be distinguished. This method required only small quantities of DEAE-Sephacel (7 mg), snAChR (0.54 micrograms), and 125I-neurotoxin (90 fmol) for each reaction, and minimized the handling of isotopic materials as compared with the conventional methods. This method is reliable, reproducible, and superior to current methods for the determination of the snAChR-binding activity for alpha-neurotoxins. PMID- 7490260 TI - Catalytic role of an arginine residue in the highly conserved and unique sequence of phosphoenolpyruvate carboxylase. AB - Phosphoenolpyruvate carboxylase (PEPC) [EC 4.1.1.31] has a highly conserved and unique sequence, 578-FHGRGGSIGRGGAP-591 (on Escherichia coli, PEPC), in which a GRGG motif is repeated twice with two intervening residues. Since previous chemical modification studies suggested the functional importance of arginine residues, the invariant Arg587 in this region was replaced with Ser, and the enzymatic properties of the resulting mutant enzyme (R587S) were investigated. Replacement led to virtual loss of the catalytic activity to form oxaloacetate. The specific activity was 37 nmol.min-1.mg-1, which corresponds to 2 x 10(-4) fold the activity of the wild-type enzyme. However, the activity of bicarbonate- and Mg(2+)-dependent hydrolysis of phosphoenolpyruvate (PEP) to pyruvate appeared for the mutant enzyme with a specific activity of 2.1 mumol.min-1.mg-1. In view of the stepwise reaction mechanism proposed for PEPC, this activity can be attributed to impairment of the subsequent partial reaction(s) following the formation of the intermediate carboxyphosphate. The half-saturation concentration (S0.5) of HCO3- in R587S was about 100-fold that in the wild-type enzyme, whereas the respective values for PEP and Mg2+ were 20- and 15-fold, indicative of this residue participating in the binding of HCO3-. PMID- 7490261 TI - Possible involvement of protein-tyrosine kinases such as p72syk in the disc sphere change response of porcine platelets. AB - We previously reported that p72syk, a non-receptor tyrosine kinase, was activated maximally at 10 s after thrombin or thromboxane A2 stimulation, even in platelets that were not allowed to aggregate [Taniguchi et al. (1993) J. Biol. Chem. 268, 2277-2279; Maeda et al. (1993) Biochem. Biophys. Res. Commun. 197, 62-67]. Then, the change in the shape of porcine platelets induced by the thromboxane A2 analogue, STA2, and the role of protein-tyrosine kinases including p72syk in this response were evaluated, using the shape-change parameter. We show that p72syk activation is correlated with the disc-sphere change in a time- and dose dependent manner following stimulation by STA2. Tyrphostin B44, a potent protein tyrosine kinase inhibitor, reduced the thromboxane A2-evoked p72syk activation and the disc-sphere change in a dose-dependent manner. Furthermore, the translocation of p72syk to the cytoskeleton-rich fraction and an increase in the tyrosine phosphorylation of an about 120 kDa protein were observed during the disc-sphere change induced by STA2. These lines of evidence suggest that the activation of protein-tyrosine kinases such as p72syk may be involved in the disc sphere change response in thromboxane A2-stimulated porcine platelets. PMID- 7490262 TI - Site-directed mutagenetic study on the role of negative patches on silene plastocyanin in the interactions with cytochrome f and photosystem I. AB - To investigate the role of two highly conserved negative patches, residues #42-45 and #59-61, on the surface of plant plastocyanin, six mutants were constructed by site-directed mutagenesis of the intermediate precursor gene from Silene pratensis. The mutants were designed systematically to incorporate positive charges into the negative patches, and the net charge on negative patches was modified from -4 to +1. Upon expression in Escherichia coli, the mutant proteins were correctly processed to the mature size and accumulated as holo-proteins. Absorption spectra, EPR, and redox potentials of the purified mutant proteins were almost indistinguishable from those of the wild-type. It was found that the electron transfer rate from cytochrome f to plastocyanin decreased exponentially as the net charge on the negative patch (#42-45) was increased, whereas the modification of the other negative patch (#59-61) had no effect. Ionic strength dependence studies indicated that the rate constants at infinite ionic strength did not change significantly among the wild-type and the six mutants, and the electrostatic attraction energies between plastocyanin and cytochrome f decreased when residues #42-45 were modified, whereas the modification of residues #59-61 had no effect. These results clearly indicated that only one (#42-45) of the two negative patches is involved in the transient complex formation with cytochrome f. Essentially similar results were observed for the electron transfer from plastocyanin to the photosystem I reaction center (P700), although in this case, slight participation of the negative patch (#59-61) is suggested.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7490263 TI - An improved phage display antibody cloning system using newly designed PCR primers optimized for Pfu DNA polymerase. AB - An improved combinatorial library system to raise mouse monoclonal antibodies was constructed. PCR primers have been newly designed to optimize the reaction for Pfu DNA polymerase, which has proofreading activity. The phagemid vector (pPDS) is designed to accommodate VH and Vk cDNAs, which had previously been assembled by PCR either in single chain fragment of variable regions (scFv) or Fab form. Antibody cloned in scFv form can be converted to Fab form by substituting the scFv linker of (Gly4Ser)3 with a fragment containing murine CH1 cDNA. This vector will produce soluble Fab in non-amber suppressor cells and allow the shuffling of light chains against a heavy chain. Hybridoma cell lines producing anti-human procollagenase monoclonal antibodies were used as the source of antibody mRNA. Antigen-binding ability of both scFv- and Fab-displaying phage was confirmed by ELISA against human procollagenase. They were also analyzed by DNA sequencing to verify the fidelity of Pfu DNA polymerase and to identify the primer incorporated. The mutation rate was considerably reduced compared to the mutation rate achieved by Taq DNA polymerase. Primers are incorporated into target sequences in most cases. PMID- 7490264 TI - Analysis of recombinant soluble mouse D-factor/LIF receptor. AB - The recombinant soluble mouse D-factor/LIF receptor (sD-factor-R) was expressed in COS-7 cells. Scatchard analysis of the bindings of mouse 125I-D-factor and human 125I-D-factor to the sD-factor-R indicated dissociation constants (Kd) of 12 and 0.267 nM, respectively, which were comparable to those of the binding protein in mouse serum. The apparent molecular masses of the sD-factor-R and human D-factor observed by gel filtration chromatography were 150 and 50 kDa, respectively. The size of the sD-factor-R.human D-factor complex was approximately 200 kDa, indicating that D-factor forms a 1:1 complex with the sD factor-R. The sD-factor-R inhibited the induction of differentiation of mouse myeloid leukemic M1 cells by mouse D-factor by blocking the binding of this factor to the cells. PMID- 7490265 TI - Phosphatidylserine-specific transbilayer lipid translocation in synaptosomal plasma membranes from Narke japonica. AB - We measured the translocation of exogenous radiolabeled phospholipid probes from the outer to the inner leaflet in the synaptosomal plasma membranes from the electric organs of Narke japonica, a Japanese marine ray. These radioactive probes can provide a convenient and highly sensitive means of measuring the translocation kinetics of phospholipids between outer and inner leaflets of biomembranes that are available only with low yield [Anzai et al. (1993) Biochim. Biophys. Acta 1151, 69-75]. Translocation kinetics revealed that the behavior of phosphatidylserine (PS), phosphatidylethanolamine (PE), and phosphatidylcholine (PC) differed. PS quickly disappeared from the outer leaflet of the bilayer. The translocation of PE and PC was slow and about 80% of the PE- and PC-probes remained in the outer leaflet at 3 h. These results differ from those of translocation in erythrocyte membranes measured using the same probes, where aminophospholipid translocase translocated both PS and PE from the outer to the inner leaflet of the bilayer, although the rate was faster for PS than for PE. In this study, the translocation was apparently PS-specific, and it was inhibited by SH reagent or intrasynaptosomal ATP-depletion. These findings clearly demonstrate that the translocation is driven by an ATP-dependent protein, which has apparent PS-specificity. We suggest that there is a translocase other than erythrocyte type aminophospholipid translocase in synaptosomal plasma membranes from N. japonica. PMID- 7490266 TI - Induction of the synthesis of hsp27 and alpha B crystallin in tissues of heat stressed rats and its suppression by ethanol or an alpha 1-adrenergic antagonist. AB - The levels of hsp27 and alpha B crystallin in various tissues of rats that had been subjected to heat stress were determined by specific immunoassays. When rats were immersed in a water bath at 42 degrees C for 20 min, the levels of hsp27 in most tissues, including central nervous tissue, liver, lung, spleen, adrenal glands, and hypophysis, had increased dramatically 8 to 16 h after the treatment. alpha B crystallin was also induced at high levels in the liver and adrenals, but not much was induced in the central nervous tissue. The increases in the levels of both hsp27 and alpha B crystallin in response to heat stress were markedly inhibited when ethanol or an alpha 1-adrenergic antagonist, prazosin, was administered before, but not after, the stress period. The expression of mRNA for hsp27 was suppressed in the livers of rats that had received ethanol or prazosin. A beta-adrenergic antagonist, propranolol, and an alpha 2-adrenergic antagonist, yohimbine, did not inhibit induction of the synthesis of the two proteins. Accumulation of hsp70 after heat stress, at least in the liver, was inhibited by ethanol or prazosin in a similar manner to that of hsp27, as detected on Western blot analysis. These results suggest that the induction of hsp27 and alpha B crystallin, as well as of hsp70, in tissues of heat-stressed rats is controlled by a physiological process(es) that is sensitive to ethanol and prazosin, and is operative for a short time during the application of heat stress. PMID- 7490267 TI - Calcium requirement and inhibitor spectrum for intracellular HIV type 1 gp160 processing in cultured HeLa cells and CD4+ lymphocytes: similarity to those of viral envelope glycoprotein maturase. AB - We recently purified the calcium-independent processing protease named viral envelope glycoprotein maturase (VEM), that converts human immunodeficiency virus type 1 (HIV-1) envelope glycoprotein precursor gp160 to gp120 and gp41, from the human CD4+ T cell line, Molt-4 clone 8 [Kido, H., Kamoshita, K., Fukutomi, A., and Katunuma, N. (1993) J. Biol. Chem. 268, 13406-13413]. In this report, we deal with the inhibitor specificity and calcium requirement for intracellular gp160 processing in cultured HeLa cells and human CD4+ lymphocytes. Processing of gp160 in these cells infected with recombinant vaccinia virus encoding the gp160 gene was not affected by intracellular calcium depletion induced by the calcium ionophore A23187 and EGTA or by intracellular calcium administration. Processing of gp160 by the purified VEM in vitro was not inhibited by EDTA, EGTA, or the metallo-protease inhibitor phosphoramidon, but was specifically inhibited by a substrate analog, decanoyl-RVKR-chloromethylketone, and the trypsin-type protease inhibitors aprotinin, HI-30, and diisopropyl fluorophosphate (DFP). It was also inhibited by E-64 and thiol reagents. But intracellular gp160 processing was inhibited only by permeable, low molecular mass inhibitors of VEM, such as DFP, E 64, and thiol reagents. Syncytium formation induced by cell surface gp120 was also inhibited by permeable inhibitors of VEM. Taken together, our results indicate that calcium ions may not be essential for intracellular gp160 processing and so HIV-1 gp160 induced by recombinant vaccinia virus may be processed mainly by a protease(s) that does not require calcium ions, such as VEM in these cells. PMID- 7490268 TI - The mechanism of interaction of sodium dodecyl sulfate with elastic fibers. AB - Sodium dodecyl sulfate (SDS), an anionic hydrophobic ligand, is known to alter the mechanical properties of elastic fibers. In order to analyze the mechanism of the alteration, two forms of fibrous elastins, "solid" and "powder" elastins, which consisted of fascicular elastic fibers and single or oligomeric elastic fibers, respectively, were prepared from bovine aorta, and the interactions of SDS with these elastins in the presence and absence of 0.15 M NaCl were studied. The solid elastin was able to retain 1.2- to 1.4-fold larger amounts of SDS than the powder elastin under both conditions, and both elastins retained 1.2-fold or larger amounts of SDS in the presence of NaCl than in its absence. Whereas both elastins released the retained SDS gradually on repeated washing with an SDS-free buffer, the release rates from the solid elastin, especially the rate in the presence of NaCl, were much smaller than those from the powder elastin, and the solid elastin retained approximately 40% of the bound SDS under conditions where the powder elastin lost most of its SDS. The SDS-binding capacities of both elastins were significantly lower than those of soluble kappa-elastin and serum albumin, which bound SDS homogeneously on the polypeptide chains. When the washed SDS-bound solid elastin was incubated with methylene blue and examined under a microscope, most of the methylene blue-SDS complex was located at the interfiber spaces of the elastic fibers. These results suggest that SDS alters the mechanical properties of elastic fibers by binding to the interfiber spaces and surfaces of the fibers rather than by binding to the internal polypeptide chains. PMID- 7490269 TI - Purification and characterization of Acinetobacter calcoaceticus 4 hydroxybenzoate 3-hydroxylase after its overexpression in Escherichia coli. AB - 4-Hydroxybenzoate 3-hydroxylase [EC 1.14.13.2] from Acinetobacter calcoaceticus was purified to homogeneity following the 40-fold overexpression of this gene (pobA) in Escherichia coli. Overexpression was accomplished by placing the folA gene (encoding trimethoprim-resistant dihydrofolate reductase) directly downstream of the pobA gene, and demanding growth of recombinants on elevated concentration of trimethoprim. Presumably, the surviving variants have undergone a genetic alteration which allowed the overexpression of both folA and pobA. 4 Hydroxybenzoate 3-hydroxylase was purified in two chromatographic steps, characterized biochemically, and its properties were compared to those of its homolog from Pseudomonas fluorescens. The two enzymes differ in their response to Cl- ion inhibition. A single amino acid change in the putative NADPH-binding site is proposed to account for this difference. The inhibitory and catalytic properties of substrate analogs were also examined. PMID- 7490270 TI - Molecular cloning and in situ localization in the brain of rat sky receptor tyrosine kinase. AB - We previously identified a human cDNA encoding a novel receptor tyrosine kinase, termed Sky, which is predominantly expressed in the brain and has a unique extracellular domain consisting of two immunoglobulin (Ig)-like and two fibronectin type III (FN III) motifs. In attempts to define the functional role of the Sky receptor, we cloned a rat sky cDNA, and localized the sites of expression of sky transcript in the adult rat brain by Northern blot and in situ hybridization analyses using the cloned rat cDNA as a probe. The deduced amino acid sequence of rat Sky has an overall sequence and a domain topology highly conserved with human Sky (90% overall identity and 98% identity within the tyrosine kinase domain). Northern blot analysis revealed that a single 3.8-kb sky mRNA is expressed in PC12 pheochromocytoma and Neuro-2a neuroblastoma cell lines and in various regions of the adult rat brain. In situ hybridization analysis revealed widespread but confined neuronal populations in adult rat brain that express sky transcript; prominent hybridization signals were detected in the inner granular layer of the olfactory bulb, CA-1 area of the hippocampus, granule cell layer of the cerebellum, tenia tectum and cingulate gyrus neurons, and wide regions of cortex layers II-VI. The high level of expression of sky mRNA in neurons in restricted brain regions suggests that the Sky receptor may play an important role in development, function, and maintenance of specific neuronal populations in the central nervous system. PMID- 7490271 TI - Aberrant expressions of decorin and biglycan genes in the carbohydrate-deficient glycoprotein syndrome. AB - Carbohydrate-deficient glycoprotein syndrome (CDGS) is a congenital disorder characterized by neurological and developmental defects. We have examined the expressions of the small proteoglycans decorin and biglycan in cultured skin fibroblasts from a patient with CDGS Type-I. Northern blotting analysis identified a marked reduction in decorin mRNA and an increase in biglycan mRNA levels. The decorin protein in the culture medium was decreased. Responses to interleukin-1 beta (IL-1 beta) and transforming growth factor-beta 1 (TGF-beta 1) were apparently abnormal; decorin was only slightly up-regulated by IL-1 beta, while biglycan was markedly down-regulated by IL-1 beta and significantly up regulated by TGF-beta 1. The constitutional and developmental abnormalities characteristic of CDGS may be associated with such derangements in the expression of proteoglycan genes. PMID- 7490272 TI - Phospholipid degradation in rat calcium ionophore-activated platelets is catalyzed mainly by two discrete secretory phospholipase As. AB - An "A1 type" phospholipase activity with serine-phospholipid preference was released by rat activated platelets. It was distinct from the secretory type II phospholipase A2 [Horigome, K., Hayakawa, M., Inoue, K., and Nojima, S. (1987) J. Biochem. 101, 625-631] and co-purified with the secretory lysophosphatidylserine selective lysophospholipase activity [Higashi, S., Kobayashi, T., Kudo, I., and Inoue, K. (1988) J. Biochem. 103, 442-447]. Several lines of evidence indicated that a single protein was responsible for the phospholipase A1 and lysophospholipase activities. Marked accumulation of lysophospholipids was observed in rat calcium ionophore-activated washed platelets and both phospholipase A1/lysophospholipase and type II phospholipase A2 were shown to contribute to this phospholipid degradation. A selective inhibitor of type II phospholipase A2 reduced the phospholipid degradation and enhanced the clotting time and prothrombinase activity. These results indicate that secretory platelet phospholipases may play a role in regulation of blood clotting. PMID- 7490273 TI - Expression and splicing of the latency-associated transcripts of herpes simplex virus type 1 in neuronal and non-neuronal cell lines. AB - Herpes simplex virus type 1 (HSV-1) is transcriptionally active during latent infection in human peripheral sensory ganglia. This transcription has been linked to the ability of the virus to reactivate, but its potential gene products and mechanisms of action are unknown. To analyze the viral latency-related transcripts in neuronal and non-neuronal cell lines in an isolated cellular system, a 10.4 kb DNA fragment, which covers the entire viral transcriptionally active latency-associated region, was cloned under control of the constitutive cytomegalovirus promoter (pNM3). During transient transfection of a human embryonic kidney 293 cell line, pNM3 expressed high levels of the 2.0 kb latency associated transcript (LAT) that was not polyadenylated. The 1.5 kb LAT as well as the minor hybridizing RNAs could not be identified by Northern blotting analysis. pNM3 expression was further analyzed following transfection of two neuronal, C1300 and ND7 cell lines. The 2.0 kb LAT was synthesized at high levels in these cell lines. The 1.5 kb LAT, which in vivo can be identified only during HSV-1 latent infection in tissues which facilitate reactivation, was present at very low amounts in 293 and C1300 cells using reverse transcription PCR analysis. Higher amounts of the 1.5 kb LAT were produced in ND7 cells, a neuronal cell line shown to possess neuronal-specific splicing proteins. However, the 1.5 kb LAT was present in ND7 cells in lesser amounts than produced during latent infection in peripheral sensory ganglia. This novel cellular system provides now a tool for future studies of the role of the 1.5 kb and the 2.0 kb LATs in HSV-1 latency. PMID- 7490274 TI - Evidence for a short form of alpha 1(IV) as a major polypeptide in bovine lens capsule. AB - The extracts from bovine lens capsule with acetic acid contained, after reduction, three major collagenous polypeptides with M(r) = 180k, 175k, and 160k, which were specifically immuno-stained with anti-type IV collagen polyclonal antibody. The biochemical properties of 180k and 160k polypeptides were akin and were distinct from that of 175k polypeptide [J. Biochem. (1993) 114,358-362]. In the present study, evidence that the 160k and 180k polypeptides from bovine lens capsule both originated from alpha 1(IV) was obtained on the basis of reactivity with a monoclonal antibody that recognizes alpha 1(IV) chain at the collagenous sequence contained in [KGEPGLPGRGFPGFP]. The epitope-bearing sequence was identified from the following three experiments. Pepsin-solubilized polypeptides from human placenta were purified by affinity chromatography on the antibody coupled column and sequenced. The restriction map of the clones positively reactive with the monoclonal antibody from human placenta cDNA library was superimposed on that of human alpha 1(IV) cDNA at a specific region. Synthetic peptides corresponding to the sequence were assayed for inhibitory activity against the reaction between epitope-bearing pepsin fragments and the antibody. The 180k and 160k polypeptides showed similar intensities in protein staining as well as in immuno-staining with the monoclonal antibody. In contrast, the 175k polypeptide did not react with the monoclonal antibody, indicating that it is a genetically distinct type IV collagen chain, presumably alpha 2(IV) from its abundance. The 160k, a major type IV collagen polypeptide, is a short form of alpha 1(IV) present as a tissue form in bovine lens capsule. PMID- 7490275 TI - The rat intrinsic factor gene: its 5'-upstream region and chief cell-specific transcription. AB - A DNA segment containing the 5'-upstream region and amino terminal reading frame of the gastric intrinsic factor gene was cloned from rat and its nucleotide sequence was determined. S1 mapping demonstrated that the transcription initiation site is located downstream of the second TATA-box sequence. Similar sequence motifs to those in the pepsinogen genes transcribed in gastric chief cells were found in the deduced sequence, suggesting that the rat intrinsic factor gene is transcribed in these cells. The genes for the intrinsic factor and its homologous protein transcobalamin I were apparently derived from a common ancestoral gene, since the positions of their intron insertions as well as the amino acid residues are conserved. Northern blot hybridization showed that the gene for the intrinsic factor is transcribed in the stomach but not detectably in the intestine, kidney, testis, brain, heart, liver, lung, or spleen. In situ hybridization using radioactive complementary RNA clearly indicated that the major transcription site in gastric glands is chief cells. Different locations of expression of intrinsic factor proteins in various mammals were observed previously using antibodies: in rat parietal cells and chief cells, in mouse chief cells, and in human parietal cells. The present results clearly demonstrated the intrinsic factor mRNA mainly in chief cells of adult rats, as in mice, suggesting that transcriptional regulation of the intrinsic factor gene is essentially the same in rodents. PMID- 7490276 TI - Mode of action of an antibacterial peptide, KLKLLLLLKLK-NH2. AB - Previously, we reported that a synthetic undecapeptide, KLKLLLLLKLK-NH2, and its D-enantiomer have potent bactericidal activities against both Gram-positive and Gram-negative bacteria. Here we examined the mode of action of KLKLLLLLKLK-NH2 with special reference to its effect on bacterial membranes. We found that both the outer and inner membrane of Escherichia coli become permeable to low molecular mass substances when treated with this peptide. Under these conditions, the bacteria lost the ability to synthesize ATP and to transport proline, suggesting that their electrochemical membrane potential was disrupted. This peptide appears to form numerous channels in bacterial membranes that interfere with membrane functions, resulting in cell death. PMID- 7490277 TI - Comprehensive site-directed mutagenesis of L-2-halo acid dehalogenase to probe catalytic amino acid residues. AB - L-2-Halo acid dehalogenase catalyzes the stereospecific hydrolytic dehalogenation of L-2-halo acids, with inversion of the C2-configuration. Seven L-2-halo acid dehalogenases from various bacterial strains are significantly similar to one another in their amino acid sequences (36-70% identity), and they are supposed to catalyze the reaction through the same mechanism. To identify catalytically important residues, we mutated all the 36 highly conserved charged and polar amino acid residues of L-2-halo acid dehalogenase from Pseudomonas sp. YL, which consists of 232 amino acid residues, by replacement of D by N, E by Q, R by K, and vice versa, S and T by A, Y and W by F, M by L, and H by N. We found that the replacement of D10, K151, S175, D180, R41, S118, T14, Y157, and N177 led to a significant loss in the enzyme activity or an increase in the Km value for the substrate, showing their involvement in the catalysis. The roles of these residues are discussed. PMID- 7490278 TI - Yeast Num1p associates with the mother cell cortex during S/G2 phase and affects microtubular functions. AB - The NUM1 gene is involved in the control of nuclear migration in Saccharomyces cerevisiae. The content of NUM1 mRNA fluctuates during the cell cycle, reaching a maximum at S/G2 phase, and the translation product Num1p associates with the cortex of mother cells mainly during S, G2, and mitosis, as seen by indirect immunofluorescence. The nuclear spindle in NUM1-deficient large-budded cells often fails to align along the mother/bud axis, while abnormally elongated astral microtubules emanate from both spindle pole bodies. A num1 null mutation confers temperature sensitivity to the cold-sensitive alpha-tubulin mutant tub1-1, and shows synthetic lethality with the beta-tubulin mutant alleles tub2-402, tub2 403, tub2-404, and tub2-405. Deletion mapping has defined three functionally important Num1p regions: a potential EF hand Ca2+ binding site, a cluster of potential phosphorylation sites and a pleckstrin homology domain. The latter domain appears to be involved in targeting Num1p to the mother cell cortex. Our data suggest that the periodically expressed NUM1 gene product controls nuclear migration by affecting astral microtubule functions. PMID- 7490279 TI - Binding of E-MAP-115 to microtubules is regulated by cell cycle-dependent phosphorylation. AB - Expression levels of E-MAP-115, a microtubule-associated protein that stabilizes microtubules, increase with epithelial cell polarization and differentiation (Masson and Kreis, 1993). Although polarizing cells contain significant amounts of this protein, they can still divide and thus all stabilized microtubules must disassemble at the onset of mitosis to allow formation of the dynamic mitotic spindle. We show here that binding of E-MAP-115 to microtubules is regulated by phosphorylation during the cell cycle. Immunolabeling of HeLa cells for E-MAP-115 indicates that the protein is absent from microtubules during early prophase and progressively reassociates with microtubules after late prophase. A fraction of E MAP-115 from HeLa cells released from a block at the G1/S boundary runs with higher apparent molecular weight on SDS-PAGE, with a peak correlating with the maximal number of cells in early stages of mitosis. E-MAP-115 from nocodazole arrested mitotic cells, which can be obtained in larger amounts, displays identical modifications and was used for further biochemical characterization. The level of incorporation of 32P into mitotic E-MAP-115 is about 15-fold higher than into the interphase protein. Specific threonine phosphorylation occurs in mitosis, and the amount of phosphate associated with serine also increases. Hyperphosphorylated E-MAP-115 from mitotic cells cannot bind stably to microtubules in vitro. These results suggest that phosphorylation of E-MAP-115 is a prerequisite for increasing the dynamic properties of the interphase microtubules which leads to the assembly of the mitotic spindle at the onset of mitosis. Microtubule-associated proteins are thus most likely key targets for kinases which control changes in microtubule dynamic properties at the G2- to M phase transition. PMID- 7490280 TI - Manganese effectively supports yeast cell-cycle progression in place of calcium. AB - Metal ion requirements for the proliferation of Saccharomyces cerevisiae were investigated. We used bis-(o-aminophenoxy)-ethane-N,N,N',N'-tetraacetic acid (BAPTA), a relatively acid tolerant chelator, to reduce the free metal ion concentrations in culture media. Chelatable metal ions were added back individually and in combination. In addition to a requirement for approximately 10 pM external free Zn2+ we found an interchangeable requirement for either 66 nM free Ca2+ or only 130 pM free Mn2+. Cells depleted of Mn2+ and Ca2+ arrested as viable cells with 2 N nuclei and tended to have very small minibuds. In the absence of added Mn2+, robust growth required approximately 60 microM total internal Ca2+. In the presence of added Mn2+, robust growth continued even when internal Ca2+ was < 3% this level. Chelator-free experiments showed that MnCl2 strongly and CaCl2 weakly restored high-temperature growth of cdc1ts strains which similarly arrest as viable cells with 2 N nuclear contents and small buds. Its much greater effectiveness compared with Ca2+ suggests that Mn2+ is likely to be a physiologic mediator of bud and nuclear development in yeast. This stands in marked contrast to a claim that Ca2+ is uniquely required for cell-cycle progression in yeast. We discuss the possibility that Mn2+ may function as an intracellular signal transducer and how this possibility relates to previous claims of Ca2+'s roles in yeast metabolism. PMID- 7490281 TI - Point mutation of adenosine triphosphate-binding motif generated rigor kinesin that selectively blocks anterograde lysosome membrane transport. AB - In the study of motor proteins, the molecular mechanism of mechanochemical coupling, as well as the cellular role of these proteins, is an important issue. To assess these questions we introduced cDNA of wild-type and site-directed mutant kinesin heavy chains into fibroblasts, and analyzed the behavior of the recombinant proteins and the mechanisms involved in organelle transports. Overexpression of wild-type kinesin significantly promoted elongation of cellular processes. Wild-type kinesin accumulated at the tips of the long processes, whereas the kinesin mutants, which contained either a T93N- or T93I mutation in the ATP-binding motif, tightly bound to microtubules in the center of the cells. These mutant kinesins could bind to microtubules in vitro, but could not dissociate from them even in the presence of ATP, and did not support microtubule motility in vitro, thereby indicating rigor-type mutations. Retrograde transport from the Golgi apparatus to the endoplasmic reticulum, as well as lysosome dispersion, was shown to be a microtubule-dependent, plus-end-directed movement. The latter was selectively blocked in the rigor-mutant cells, although the microtubule minus-end-directed motion of lysosomes was not affected. We found the point mutations that make kinesin motor in strong binding state with microtubules in vitro and showed that this mutant causes a dominant effect that selectively blocks anterograde lysosome membrane transports in vivo. PMID- 7490282 TI - Differential targeting of protein kinase C and CaM kinase II signalings to vimentin. AB - Hydrolysis of inositol phospholipids by receptor stimulation activates two separate signaling pathways, one leading to the activation of protein kinase C (C kinase) via formation of diacylglycerol. The other is the inositol trisphosphate (IP3)/Ca2+ pathway and a major downstream kinase which is activated is Ca2+/calmodulin-dependent protein kinase II (CaM kinase II). To examine signaling pathways of C kinase and CaM kinase II to the cytoskeletal protein vimentin, we prepared monoclonal antibodies YT33 and MO82 which recognize the phosphorylation state of vimentin by C kinase and by CaM kinase II, respectively. Ectopic expression of constitutively active C kinase or CaM kinase II in primary cultured astrocytes by microinjection of the corresponding expression vectors induced phosphorylation of vimentin at each specific phosphorylation site, followed by reorganization of vimentin filament networks. In contrast, simultaneous activation of C kinase and CaM kinase II by inositol phospholipid hydrolysis with receptor stimulation led to an exclusive phosphorylation of vimentin at the CaM kinase II site, not at the site of C kinase. These results indicate that the intracellular targeting of C kinase and CaM kinase II signalings to vimentin is regulated separately, under physiological conditions. PMID- 7490283 TI - Binding between the neural cell adhesion molecules axonin-1 and Nr-CAM/Bravo is involved in neuron-glia interaction. AB - Neural cell adhesion molecules of the immunoglobulin superfamily mediate cellular interactions via homophilic binding to identical molecules and heterophilic binding to other family members or structurally unrelated cell-surface glycoproteins. Here we report on an interaction between axonin-1 and Nr CAM/Bravo. In search for novel ligands of axonin-1, fluorescent polystyrene microspheres conjugated with axonin-1 were found to bind to peripheral glial cells from dorsal root ganglia. By antibody blockage experiments an axonin-1 receptor on the glial cells was identified as Nr-CAM. The specificity of the interaction was confirmed with binding studies using purified axonin-1 and Nr CAM. In cultures of dissociated dorsal root ganglia antibodies against axonin-1 and Nr-CAM perturbed the formation of contacts between neurites and peripheral glial cells. Together, these results implicate a binding between axonin-1 of the neuritic and Nr-CAM of the glial cell membrane in the early phase of axon ensheathment in the peripheral nervous system. PMID- 7490284 TI - Thrombospondin-4, an extracellular matrix protein expressed in the developing and adult nervous system promotes neurite outgrowth. AB - Extracellular matrix (ECM) molecules are involved in multiple aspects of cell-to cell signaling during development and in the adult. In nervous system development, specific recognition processes, e.g., during axonal pathfinding and synaptogenesis involve modulation and signaling by ECM components. Much less is known about their presence and possible roles in the adult nervous system. We now report that thrombospondin-4 (TSP-4), a recently discovered member of the TSP gene family is expressed by neurons, promotes neurite outgrowth, and accumulates at the neuromuscular junction and at certain synapse-rich structures in the adult. To search for muscle genes that may be involved in neuromuscular signaling, we isolated cDNAs induced in adult skeletal muscle by denervation. One of these cDNAs coded for the rat homologue of TSP-4. In skeletal muscle, it was expressed by muscle interstitial cells. The transcript was further detected in heart and in the developing and adult nervous system, where it was expressed by a wide range of neurons. An antiserum to the unique carboxyl-terminal end of the protein allowed to specifically detect TSP-4 in transfected cells in vitro and on cryostat sections in situ. TSP-4 associated with ECM structures in vitro and in vivo. In the adult, it accumulated at the neuromuscular junction and at synapse rich structures in the cerebellum and retina. To analyze possible activities of TSP-4 towards neurons, we carried out coculture experiments with stably transfected COS cells and motor, sensory, or retina neurons. These experiments revealed that TSP-4 was a preferred substrate for these neurons, and promoted neurite outgrowth. The results establish TSP-4 as a neuronal ECM protein associated with certain synapse-rich structures in the adult. Its activity towards embryonic neurons in vitro and its distribution in vivo suggest that it may be involved in local signaling in the developing and adult nervous system. PMID- 7490285 TI - Glucocorticoid and progesterone inhibit involution and programmed cell death in the mouse mammary gland. AB - Milk production during lactation is a consequence of the suckling stimulus and the presence of glucocorticoids, prolactin, and insulin. After weaning the glucocorticoid hormone level drops, secretory mammary epithelial cells die by programmed cell death and the gland is prepared for a new pregnancy. We studied the role of steroid hormones and prolactin on the mammary gland structure, milk protein synthesis, and on programmed cell death. Slow-release plastic pellets containing individual hormones were implanted into a single mammary gland at lactation. At the same time the pups were removed and the consequences of the release of hormones were investigated histologically and biochemically. We found a local inhibition of involution in the vicinity of deoxycorticosterone- and progesterone-release pellets while prolactin-release pellets were ineffective. Dexamethasone, a very stable and potent glucocorticoid hormone analogue, inhibited involution and programmed cell death in all the mammary glands. It led to an accumulation of milk in the glands and was accompanied by an induction of protein kinase A, AP-1 DNA binding activity and elevated c-fos, junB, and junD mRNA levels. Several potential target genes of AP-1 such as stromelysin-1, c-jun, and SGP-2 that are induced during normal involution were strongly inhibited in dexamethasone-treated animals. Our results suggest that the cross-talk between steroid hormone receptors and AP-1 previously described in cells in culture leads to an impairment of AP-1 activity and to an inhibition of involution in the mammary gland implying that programmed cell death in the postlactational mammary gland depends on functional AP-1. PMID- 7490287 TI - Mutational analysis of p80 coilin indicates a functional interaction between coiled bodies and the nucleolus. AB - Coiled bodies are conserved subnuclear domains found in both plant and animal cells. They contain a subset of splicing snRNPs and several nucleolar antigens, including Nopp140 and fibrillarin. In addition, autoimmune patient sera have identified a coiled body specific protein, called p80 coilin. In this study we show that p80 coilin is ubiquitously expressed in human tissues. The full-length human p80 coilin protein correctly localizes in coiled bodies when exogenously expressed in HeLa cells using a transient transfection assay. Mutational analysis identifies separate domains in the p80 coilin protein that differentially affect its subnuclear localization. The data show that p80 coilin has a nuclear localization signal, but this is not sufficient to target the protein to coiled bodies. The results indicate that localization in coiled bodies is not determined by a simple motif analogous to the NLS motifs involved in nuclear import. A specific carboxy-terminal deletion in p80 coilin results in the formation of pseudo-coiled bodies that are unable to recruit splicing snRNPs. This causes a loss of endogenous coiled bodies. A separate class of mutant coilin proteins are shown to localize in fibrillar structures that surround nucleoli. These mutants also lead to loss of endogenous coiled bodies, produce a dramatic disruption of nucleolar architecture and cause a specific segregation of nucleolar antigens. The structural change in nucleoli is accompanied by the loss of RNA polymerase I activity. These data indicate that p80 coilin plays an important role in subnuclear organization and suggest that there may be a functional interaction between coiled bodies and nucleoli. PMID- 7490286 TI - Testicular development involves the spatiotemporal control of PDGFs and PDGF receptors gene expression and action. AB - Platelet-derived growth factors (PDGFs) are growth-regulatory molecules that stimulate chemotaxis, proliferation and metabolism primarily of cells of mesenchymal origin. In this study, we found high levels of PDGFs and PDGFs receptors (PDGFRs) mRNAs, and specific immunostaining for the corresponding proteins in the rat testis. PDGFs and PDGFRs expression was shown to be developmentally regulated and tissue specific. Expression of PDGFs and PDGFRs genes was observed in whole testis RNA 2 d before birth, increased through postnatal day 5 and fell to low levels in adult. The predominant cell population expressing transcripts of the PDGFs and PDGFRs genes during prenatal and early postnatal periods were Sertoli cells and peritubular myoid cells (PMC) or their precursors, respectively, while in adult animals PDGFs and PDGFRs were confined in Leydig cells. We also found that early postnatal Sertoli cells produce PDGF like substances and that this production is inhibited dose dependently by follicle-stimulating hormone (FSH). The expression of PDGFRs by PMC and of PDGFs by Sertoli cells corresponds in temporal sequence to the developmental period of PMC proliferation and migration from the interstitium to the peritubulum. Moreover, we observed that all the PDGF isoforms and the medium conditioned by early postnatal Sertoli cells show a strong chemotactic activity for PMC which is inhibited by anti-PDGF antibodies. These data indicate that, through the spatiotemporal pattern of PDGF ligands and receptors expression, PDGF may play a role in testicular development and homeostasis. PMID- 7490288 TI - Identification of the chromosome localization domain of the Drosophila nod kinesin-like protein. AB - The nod kinesin-like protein is localized along the arms of meiotic chromosomes and is required to maintain the position of achiasmate chromosomes on the developing meiotic spindle. Here we show that the localization of ectopically expressed nod protein on mitotic chromosomes precisely parallels that observed for wild-type nod protein on meiotic chromosomes. Moreover, the carboxyl-terminal half of the nod protein also binds to chromosomes when overexpressed in mitotic cells, whereas the overexpressed amino-terminal motor domain binds only to microtubules. Chromosome localization of the carboxyl-terminal domain of nod depends upon an 82-amino acid region comprised of three copies of a sequence homologous to the DNA-binding domain of HMG 14/17 proteins. These data map the two primary functional domains of the nod protein in vivo and provide a molecular explanation for the directing of the nod protein to a specific subcellular component, the chromosome. PMID- 7490290 TI - FAR1 is required for oriented polarization of yeast cells in response to mating pheromones. AB - Cell polarization involves specifying an area on the cell surface and organizing the cytoskeleton towards that landmark. The mechanisms by which external signals are translated into internal landmarks for polarization are poorly understood. The yeast Saccharomyces cerevisiae exhibits polarized growth during mating: the actin cytoskeleton of each cell polarizes towards its partner, presumably to allow efficient cell fusion. The external signal which determines the landmark for polarization is thought to be a gradient of peptide pheromone released by the mating partner. Here we described mutants that exhibit random polarization. Using two assays, including a direct microscope assay for orientation (Segall, J. 1993. Proc. Natl. Acad. Sci. USA. 90:8332-8337), we show that these mutants cannot locate the source of a pheromone gradient although they are able to organize their cytoskeleton. These mutants appear to be defective in mating because they are unable to locate the mating partner. They carry mutations of the FAR1 gene, denoted far1-s, and identify a new function for the Far1 protein. Its other known function is to promote cell cycle arrest during mating by inhibiting a cyclin dependent kinase (Peter, M., and I. Herskowitz. 1994. Science (Wash. DC). 265:1228-1232). The far1-s mutants exhibit normal cell cycle arrest in response to pheromone, which suggests that Far1 protein plays two distinct roles in mating: one in cell cycle arrest and the other in orientation towards the mating partner. PMID- 7490289 TI - Saccharomyces cerevisiae cells execute a default pathway to select a mate in the absence of pheromone gradients. AB - During conjugation, haploid S. cerevisiae cells find one another by polarizing their growth toward each other along gradients of pheromone (chemotropism). We demonstrate that yeast cells exhibit a second mating behavior: when their receptors are saturated with pheromone, wild-type a cells execute a default pathway and select a mate at random. These matings are less efficient than chemotropic matings, are induced by the same dose of pheromone that induces shmoo formation, and appear to use a site near the incipient bud site for polarization. We show that the SPA2 gene is specifically required for the default pathway: spa2 delta mutants cannot mate if pheromone concentrations are high and gradients are absent, but can mate if gradients are present. ste2 delta, sst2 delta, and far1 delta mutants are chemotropism-defective and therefore must choose a mate by using a default pathway; consistent with this deduction, these strains require SPA2 to mate. In addition, our results suggest that far1 mutants are chemotropism defective because their mating polarity is fixed at the incipient bud site, suggesting that the FAR1 gene is required for inhibiting the use of the incipient bud site during chemotropic mating. These observations reveal a molecular relationship between the mating and budding polarity pathways. PMID- 7490291 TI - Sequential coupling between COPII and COPI vesicle coats in endoplasmic reticulum to Golgi transport. AB - COPI and COPII are vesicle coat complexes whose assembly is regulated by the ARF1 and Sar1 GTPases, respectively. We show that COPI and COPII coat complexes are recruited separately and independently to ER (COPII), pre-Golgi (COPI, COPII), and Golgi (COPI) membranes of mammalian cells. To address their individual roles in ER to Golgi transport, we used stage specific in vitro transport assays to synchronize movement of cargo to and from pre-Golgi intermediates, and GDP- and GTP-restricted forms of Sar1 and ARF1 proteins to control coat recruitment. We find that COPII is solely responsible for export from the ER, is lost rapidly following vesicle budding and mediates a vesicular step required for the build-up of pre-Golgi intermediates composed of clusters of vesicles and small tubular elements. COPI is recruited onto pre-Golgi intermediates where it initiates segregation of the anterograde transported protein vesicular stomatitis virus glycoprotein (VSV-G) from the retrograde transported protein p58, a protein which actively recycles between the ER and pre-Golgi intermediates. We propose that sequential coupling between COPII and COPI coats is essential to coordinate and direct bi-directional vesicular traffic between the ER and pre-Golgi intermediates involved in transport of protein to the Golgi complex. PMID- 7490292 TI - The Golgi-localization of yeast Emp47p depends on its di-lysine motif but is not affected by the ret1-1 mutation in alpha-COP. AB - The Saccharomyces cerevisiae EMP47 gene encodes a nonessential type-I transmembrane protein with sequence homology to a class of intracellular lectins defined by ERGIC-53 and VIP36. The 12-amino acid COOH-terminal cytoplasmic tail of Emp47p ends in the sequence KTKLL, which conforms with the consensus for di lysine-based ER-localization signals. Despite the presence of this motif, Emp47p was shown to be a Golgi protein at steady-state. The di-lysine motif of Emp47p was functional when transplanted onto Ste2p, a plasma membrane protein, conferring ER localization. Nevertheless, the di-lysine motif was required for Golgi-localization of Emp47p and showed the same charge-independent, position dependent characteristics of other di-lysine motifs. Alpha-COP has been shown to be required for ER localization of di-lysine-tagged proteins. Consistent with this finding, the Ste2p-Emp47p hybrid protein was mislocalized to the cell surface in the alpha-COP mutant, ret1-1. Surprisingly, the Golgi-localization of Emp47p was unaffected by the ret1-1 mutation. To investigate whether Emp47p undergoes retrograde transport from the Golgi to the ER like other di-lysine tagged proteins we developed an assay to measure this step after block of forward transport in a sec12 mutant. Under these conditions retrograde transport led to a specific redistribution of Emp47p from the Golgi to the ER. This recycling occurred from a Golgi subcompartment containing alpha 1,3 mannose-modified oligosaccharides suggesting that it originated from a medial-or later Golgi compartment. Thus Emp47p cycles between the Golgi apparatus and the ER and requires a di-lysine motif for its alpha-COP-independent, steady state localization in the Golgi. PMID- 7490293 TI - Localization and targeting of the Saccharomyces cerevisiae Kre2p/Mnt1p alpha 1,2 mannosyltransferase to a medial-Golgi compartment. AB - The yeast Kre2p/Mnt1p alpha 1,2-mannosyltransferase is a type II membrane protein with a short cytoplasmic amino terminus, a membrane-spanning region, and a large catalytic luminal domain containing one N-glycosylation site. Anti-Kre2p/Mnt1p antibodies identify a 60-kD integral membrane protein that is progressively N glycosylated in an MNN1-dependent manner. Kre2p/Mnt1p is localized in a Golgi compartment that overlaps with that containing the medial-Golgi mannosyltransferase Mnn1p, and distinct from that including the late Golgi protein Kex1p. To determine which regions of Kre2p/Mnt1p are required for Golgi localization, Kre2p/Mnt1p mutant proteins were assembled by substitution of Kre2p domains with equivalent sequences from the vacuolar proteins DPAP B and Pho8p. Chimeric proteins were tested for correct topology, in vitro and in vivo activity, and were localized intracellularly by indirect immunofluorescence. The results demonstrate that the NH2-terminal cytoplasmic domain is necessary for correct Kre2p Golgi localization whereas, the membrane-spanning and stem domains are dispensable. However, in a test of targeting sufficiency, the presence of the entire Kre2p cytoplasmic tail, plus the transmembrane domain and a 36-amino acid residue luminal stem region was required to localize a Pho8p reporter protein to the yeast Golgi. PMID- 7490294 TI - Hormonal regulation of caveolae internalization. AB - Caveolae undergo a cyclic transition from a flat segment of membrane to a vesicle that then returns to the cell surface. Here we present evidence that this cycle depends on a population of protein kinase C-alpha (PKC-alpha) molecules that reside in the caveolae membrane where they phosphorylate a 90-kD protein. This cycle can be interrupted by treatment of the cells with phorbol-12,13-dibutyrate or agents that raise the concentration of diacylglycerol in the cell. Each of these conditions displaces PKC-alpha from caveolae, inhibits the phosphorylation of the 90-kD protein, and prevents internalization. Caveolae also contain a protein phosphatase that dephosphorylates the 90-kD once PKC-alpha is gone. A similar dissociation of PKC-alpha from caveolae and inhibition of invagination was observed when cells were treated with histamine. This effect was blocked by pyrilamine but not cimetidine, indicating the involvement of histamine H1 receptors. These findings suggest that the caveolae internalization cycle is hormonally regulated. PMID- 7490295 TI - A transferrin-like GPI-linked iron-binding protein in detergent-insoluble noncaveolar microdomains at the apical surface of fetal intestinal epithelial cells. AB - A GPI-anchored 80-kD protein was found to be the major component of detergent insoluble complexes, prepared from fetal porcine small intestine, constituting about 25% of the total amount of protein. An antibody was raised to the 80-kD protein, and by immunogold electron microscopy of ultracryosections of mucosal tissue, the protein was localized to the apical surface of the enterocytes, whereas it was absent from the basolateral plasma membrane. Interestingly, it was mainly found in patches of flat or invaginated apical membrane domains rather than at the surface of microvilli. Caveolae were not found in association with these labeled microdomains. In addition, the 80-kD protein was seen in apical endocytic vacuoles and in tubulo-vesicular structures, suggesting that the apical microdomains are involved in endocytosis of the 80-kD protein. By its NH2 terminal amino acid sequence, iron-binding capacity and partial immunological cross-reactivity with serum transferrin, the 80-kD protein was shown to belong to the transferrin family, and it is probably homologous to melanotransferrin, a human melanoma-associated antigen. The 80-kD iron-binding protein was fully detergent-soluble immediately after synthesis and only became insoluble after gaining resistance to endo H, supporting a mechanism for exocytic delivery to the apical cell surface by way of detergent-insoluble glycolipid "rafts" that fuse with the plasmalemma at restricted sites devoid of microvilli. PMID- 7490297 TI - Regulatory mechanisms of the acrosome reaction revealed by multiview microscopy of single starfish sperm. AB - The acrosome reaction in many animals is a coupled reaction involving an exocytotic step and a dramatic change in cell shape. It has been proposed that these morphological changes are regulated by intracellular ions such as Ca2+ and H+. We report here simultaneous visualization, under a multiview microscope, of intracellular free Ca2+ concentration ([Ca2+]i), intracellular pH (pHi), and morphological changes in a single starfish sperm (Asterina pectinifera). [Ca2+]i and pHi were monitored with the fluorescent probes indo-1 and SNARF-1, respectively. The acrosome reaction was induced with ionomycin. After the introduction of ionomycin in the medium, [Ca2+]i increased gradually and reached a plateau in approximately 30 s. The fusion of the acrosomal vacuole took place abruptly before the plateau, during the rising phase. Although the speed of the [Ca2+]i increase varied among the many sperm tested, exocytosis in all cases occurred at the same [Ca2+]i of approximately 2 microM (estimated using the dissociation constant of indo-1 for Ca2+ of 1.1 microM). This result suggests that the exocytotic mechanism in starfish sperm responds to [Ca2+]i rapidly, with a reaction time of the order of one second or less. Unlike the change in [Ca2+]i, an abrupt increase in pHi was observed immediately after exocytosis, suggesting the presence of a proton mobilizing system that is triggered by exocytosis. The rapid increase in pHi coincided with the formation of the acrosomal rod and the beginning of vigorous movement of the flagellum, both of which have been proposed to be pHi dependent. The exocytotic event itself was visualized with the fluorescent membrane probe RH292. The membrane of the acrosomal vacuole, concealed from the external medium in an unreacted sperm, was seen to fuse with the plasma membrane. PMID- 7490298 TI - Successful histocompatible myoblast transplantation in dystrophin-deficient mdx mouse despite the production of antibodies against dystrophin. AB - Myoblast transplantation has been considered a potential treatment for some muscular disorders. It has proven very successful, however, only in immunodeficient or immunosuppressed mice. In this study, myoblasts from C57BL10J +/+ mice were transplanted, with no immunosuppressive treatment, in the tibialis anterior of fully histocompatible but dystrophin-deficient C57BL10J mdx/mdx mice. One to 9 months after transplantation, the success of the graft was evaluated by immunohistochemistry. All the transplanted mice (n = 24) developed dystrophin positive fibers following transplantation. Depending on myoblast cultures, transplantations, and time of analysis, the mice presented 15 to 80% of dystrophin-positive fibers in transplanted muscles. These fibers were correctly oriented and they were either from donor or hybrid origin. The dystrophin positive fibers remained stable up to 9 months. Possible humoral and cellular immune responses were investigated after grafting. Antibodies directed against dystrophin and/or muscle membrane were developed by 58% of the mice as demonstrated by immunohistochemistry and Western blotting. Despite the presence of these antibodies, dystrophin-positive fibers were still present in grafted muscles 9 months after transplantation. Moreover, the muscles did not show massive infiltration by CD4 cells, CD8 cells, or macrophages, as already described in myoblast allotransplantations. This lack of rejection was attributed to the sequestrated nature of dystrophin after fiber formation. These results indicate that myoblast transplantation leads to fiber formation when immunocompetent but fully histocompatible donors and recipients are used and that dystrophin incompatibility alone is not sufficient to induce an immunological rejection reaction. PMID- 7490299 TI - Myosin II filament assemblies in the active lamella of fibroblasts: their morphogenesis and role in the formation of actin filament bundles. AB - The morphogenesis of myosin II structures in active lamella undergoing net protrusion was analyzed by correlative fluorescence and electron microscopy. In rat embryo fibroblasts (REF 52) microinjected with tetramethylrhodamine-myosin II, nascent myosin spots formed close to the active edge during periods of retraction and then elongated into wavy ribbons of uniform width. The spots and ribbons initially behaved as distinct structural entities but subsequently aligned with each other in a sarcomeric-like pattern. Electron microscopy established that the spots and ribbons consisted of bipolar minifilaments associated with each other at their head-containing ends and arranged in a single row in an "open" zig-zag conformation or as a "closed" parallel stack. Ribbons also contacted each other in a nonsarcomeric, network-like arrangement as described previously (Verkhovsky and Borisy, 1993. J. Cell Biol. 123:637-652). Myosin ribbons were particularly pronounced in REF 52 cells, but small ribbons and networks were found also in a range of other mammalian cells. At the edge of the cell, individual spots and open ribbons were associated with relatively disordered actin filaments. Further from the edge, myosin filament alignment increased in parallel with the development of actin bundles. In actin bundles, the actin cross-linking protein, alpha-actinin, was excluded from sites of myosin localization but concentrated in paired sites flanking each myosin ribbon, suggesting that myosin filament association may initiate a pathway for the formation of actin filament bundles. We propose that zig-zag assemblies of myosin II filaments induce the formation of actin bundles by pulling on an actin filament network and that co-alignment of actin and myosin filaments proceeds via folding of myosin II filament assemblies in an accordion-like fashion. PMID- 7490302 TI - Recognition and empowerment: an effective approach to enlisting patient cooperation. PMID- 7490301 TI - Attachment corrosion and its testing. PMID- 7490303 TI - Spontaneous correction of a mesially inclined canine. PMID- 7490300 TI - Enantiomeric separation of diltiazem, clentiazem, and its related compounds by capillary electrophoresis using polysaccharides. AB - The direct separation of the enantiomers of diltiazem and its three chloro derivatives, including clentiazem, was investigated by capillary zone electrophoresis (CZE) and electrokinetic chromatography (EKC) using polysaccharides. An electrically neutral beta-cyclodextrin polymer, dextran sulfate, and mucopolysaccharides, such as heparin and chondroitin sulfate C, were employed as chiral selectors. Among the tested solutes, 8-chlorodiltiazem (clentiazem) was well enantiorecognized by the methods. Affinity electrokinetic chromatography (AEKC) using chondroitin sulfate C was the most successful; all tested solutes were completely enantioseparated by this mode. PMID- 7490296 TI - Targeting of cholera toxin and Escherichia coli heat labile toxin in polarized epithelia: role of COOH-terminal KDEL. AB - Vibrio cholerae and Escherichia coli heat labile toxins (CT and LT) elicit a secretory response from intestinal epithelia by binding apical receptors (ganglioside GM1) and subsequently activating basolateral effectors (adenylate cyclase). We have recently proposed that signal transduction in polarized cells may require transcytosis of toxin-containing membranes (Lencer, W. I., G. Strohmeier, S. Moe, S. L. Carlson, C. T. Constable, and J. L. Madara. 1995. Proc. Natl. Acad. Sci. USA. 92:10094-10098). Targeting of CT into this pathway depends initially on binding of toxin B subunits to GM1 at the cell surface. The anatomical compartments in which subsequent steps of CT processing occur are less clearly defined. However, the enzymatically active A subunit of CT contains the ER retention signal KDEL (RDEL in LT). Thus if the KDEL motif were required for normal CT trafficking, movement of CT from the Golgi to ER would be implied. To test this idea, recombinant wild-type (wt) and mutant CT and LT were prepared. The COOH-terminal KDEL sequence in CT was replaced by seven unrelated amino acids: LEDERAS. In LT, a single point mutation replacing leucine with valine in RDEL was made. Wt and mutant toxins displayed similar enzymatic activities and binding affinities to GM1 immobilized on plastic. Biologic activity of recombinant toxins was assessed as a Cl- secretory response elicited from the polarized human epithelial cell line T84 using standard electrophysiologic techniques. Mutations in K(R)DEL of both CT and LT delayed the time course of toxin-induced Cl- secretion. At T1/2, dose dependencies for K(R)DEL-mutant toxins were increased > or = 10-fold. KDEL-mutants displayed differentially greater temperature sensitivity. In direct concordance with a slower rate of signal transduction. KDEL-mutants were trafficked to the basolateral membrane more slowly than wt CT (assessed by selective cell surface biotinylation as transcytosis of B subunit). Mutation in K(R)DEL had no effect on the rate of toxin endocytosis. These data provide evidence that CT and LT interact directly with endogenous KDEL-receptors and imply that both toxins may require retrograde movement through Golgi cisternae and ER for efficient and maximal biologic activity. PMID- 7490304 TI - Risk management. Additional fee in case of non-cooperation in orthodontics. PMID- 7490305 TI - Light and setting times of visible-light-cured orthodontic adhesives. PMID- 7490306 TI - Third-generation mandibular bonded lingual 3-3 retainer. PMID- 7490307 TI - Instructions for wearing elastics. PMID- 7490309 TI - Computerised image handling in pathology. PMID- 7490308 TI - Molecular genetics of familial breast-ovarian cancer. PMID- 7490310 TI - Detection of C-polysaccharide in serum of patients with Streptococcus pneumoniae bacteraemia. AB - AIM: To investigate the fate of Streptococcus pneumoniae C-polysaccharide antigen in serum in patients with S pneumoniae bacteraemia. METHOD: In vitro dissociation experiments were performed to demonstrate that C-polysaccharide was masked by ligands in normal and acute phase serum. Serum samples from 22 patients with S pneumoniae bacteraemia were treated to dissociate immune complexes and then tested for C-polysaccharide by enzyme linked immunosorbent assay (ELISA). RESULTS: C-polysaccharide antigen was masked in normal and acute phase serum but could be released by EDTA treatment and detected by ELISA. Antigen was found in six patients ranging in concentration from 2.5 to 200 ng/ml. Patients with detectable antigen were more likely to die than those in whom antigen was not detected. CONCLUSION: This study demonstrates that C-polysaccharide antigen commonly circulates in patients with S pneumoniae bacteraemia but its presence is masked by ligands present in serum. PMID- 7490311 TI - Evaluation of the AUXACOLOR system, a new method of clinical yeast identification. AB - AIMS: To compare the AUXACOLOR yeast identification system with the API 20C system. METHODS: Yeast isolates (n = 215), comprising 16 species, were identified using the AUXACOLOR system and the API 20C system. Isolates that could not be identified with the API 20C system or which produced discrepant results in the two systems were identified by assimilation and fermentation procedures. RESULTS: AUXACOLOR correctly identified 150 (85.7%) of 175 germ tube negative isolates while API 20C identified 155 (88.6%). Incorrect identifications were more common with API 20C (7.4%) than with AUXACOLOR (3.7%). Of 110 isolates of four common pathogens (Candida glabrata, C parapsilosis, C tropicalis, and Cryptococcus neoformans), 82.7% (91/110) were identified by AUXACOLOR while API 20C identified 74.5% (82/110). Of 65 less common germ tube negative isolates, 55.4% (36/65) were identified by AUXACOLOR while API 20C identified 63.1% (41/65). CONCLUSION: Although it has a limited database of 26 species, the AUXACOLOR system is a useful method for identification of germ tube negative clinical yeast isolates. Compared with the API 20C, the AUXACOLOR system is simpler and quicker to set up, easier to interpret, and comparable in cost. PMID- 7490312 TI - Routine application of the polymerase chain reaction for detection of Mycobacterium tuberculosis in clinical samples. AB - AIM: To investigate the use of the polymerase chain reaction (PCR) in the routine laboratory for the detection of Mycobacterium tuberculosis in clinical samples. METHODS: Samples were divided and processed separately for the detection of M tuberculosis by microscopy, culture and PCR. After DNA extraction, PCR was performed with primers specific for the insertion element IS6110 and the product was analysed by agarose gel electrophoresis, Southern blotting or dot blotting and hybridisation with a digoxigenin labelled internal probe. Each sample was tested for inhibitors of Taq polymerase with the aid of an internal control. Multiple negative and positive controls were used to monitor each step of the procedure. RESULTS: The data from two laboratories, using the same operating procedures, were combined. Of 1957 specimens, 79 (4%) were culture and PCR positive, while 1839 (93.9%) were negative in both tests. Thirty specimens (1.5%) were PCR positive only and nine (0.5%) were culture positive but PCR negative. CONCLUSION: Using culture and clinical history as the gold standard, sensitivity and specificity for PCR were 92.1% and 99.8%, respectively. With elaborate precautions, PCR is a suitable and reliable method for the detection of M tuberculosis in clinical samples in a routine microbiology laboratory. PMID- 7490313 TI - Chlamydia trachomatis and ectopic pregnancy: retrospective analysis of salpingectomy specimens, endometrial biopsies, and cervical smears. AB - AIMS: To examine the role of Chlamydia trachomatis in ectopic pregnancy by detection of DNA in archival salpingectomy specimens, and in their preceding cervical specimens and endometrial biopsies, by using the polymerase chain reaction (PCR). METHODS: Archival paraffin embedded salpingectomy tissues (n = 48) from 37 women with ectopic pregnancy were examined for the presence of C trachomatis plasmid and omp1 DNA by PCR. In addition, preceding cervical specimens (n = 58) stored either as cervical cell suspensions or as archival cervical smears, and preceding endometrial biopsies (n = 18), taken 0-5.8 years before the ectopic pregnancy, were examined by PCR for the presence of C trachomatis. RESULTS: C trachomatis DNA was detected in only one of the 48 salpingectomy specimens from 37 women. However, in six of the 37 women, C trachomatis DNA was detected in the genital specimens (cervix and/or endometrial) taken before salpingectomy. C trachomatis infections were mostly found in endometrial or cervical specimens taken more than three years before ectopic pregnancy. No chlamydial DNA was found in endometrial or cervical specimens taken at the same time of the ectopic pregnancy. CONCLUSIONS: Although no C trachomatis DNA was found in salpingectomy specimens, several women with ectopic pregnancy had C trachomatis infections in endometrial and cervical specimens in the past. This suggests that at least in these cases the ectopic pregnancy is a late post inflammatory complication of an ascending C trachomatis infection resulting in a scarred fallopian tube. PMID- 7490314 TI - Quantitation of hepatitis C viral RNA in liver and serum samples using competitive polymerase chain reaction. AB - AIMS: To investigate whether the amount of hepatitis C viral RNA (HCV-RNA) in liver and serum can predict the effectiveness of interferon treatment in patients with chronic hepatitis C. METHODS: The amount of HCV-RNA in frozen liver tissues and sera of 22 patients with chronic hepatitis C was determined before and after interferon alfa treatment by the competitive reverse transcriptional polymerase chain reaction method. RESULTS: Patients with small amounts of HCV-RNA in serum before treatment showed a significantly more effective response to interferon treatment. After treatment, HCV-RNA disappeared from both the liver and serum of all patients who sustained complete response (n = 11); in contrast, HCV-RNA decreased but persisted in the liver of all those who relapsed after cessation of treatment (n = 11). CONCLUSIONS: The elimination of hepatitis C virus from the liver as well as from the serum seems to be essential for sustained remission. The quantitation of HCV-RNA in liver biopsy specimens obtained after treatment would be a highly accurate predictor of whether relapse is likely to occur. PMID- 7490316 TI - Identification of parietal cells in gastric body mucosa with HMFG-2 monoclonal antibody. AB - AIMS: To identify parietal cells in the upper gastrointestinal tract by an immunoperoxidase method, using commercially available monoclonal antibodies. METHODS: Routine surgical biopsy specimens of gastric body mucosa were examined using the avidin-biotin peroxidase method with the monoclonal antibodies HMFG-1 and HMFG-2 to identify parietal cells. Double immunoperoxidase labelling with HK12.18, a well characterised monoclonal antibody directed against an epitope on the alpha (catalytic) subunit of H+ translocating, K+ stimulated adenosine triphosphatase (H,K-ATPase), was used to confirm that HMFG-1 and -2 stained parietal cells. RESULTS: HMFG-1 and HMFG-2 showed consistent parietal cell staining patterns in the gastric body mucosa. HMFG-2 gave a more intense staining pattern of the secretory canaliculi. This was confirmed by double immunolabelling with HK12.18. CONCLUSIONS: HMFG monoclonal antibodies are recommended as highly specific markers of human gastric parietal cells. PMID- 7490318 TI - Immunohistochemical staining of plastic embedded bone marrow trephine biopsy specimens after microwave heating. AB - AIMS: To investigate (1) whether adequate immunohistochemical staining can be achieved on sections cut from plastic embedded bone marrow trephine biopsy specimens after microwave heating in citrate buffer; and (2) whether this immunohistochemical staining is comparable with that achieved on routine sections cut from paraffin wax embedded trephine biopsy specimens after decalcification procedures. METHODS: Sixty five consecutive bone marrow trephine biopsy specimens of more than 1 cm in length were divided transversely into two equal parts. One part was processed in paraffin wax followed by decalcification. The other part was embedded in the epoxyresin Polarbed 812 followed by the cutting of 1 micron sections. Both parts underwent immunohistochemical staining by an identical panel of antibodies. With Polarbed 812 plastic embedded sections, microwave heating in citrate buffer was undertaken before the application of antisera. RESULTS: On sections cut from plastic embedded material, immunohistochemical staining was generally satisfactory, easy to interpret and comparable with that achieved with paraffin wax embedded material. Exceptions were antibodies to neutrophil elastase and CD61 where immunostaining was consistently negative on plastic embedded sections. Immunohistochemical staining for CD20 was consistently more reliable on plastic embedded sections. CONCLUSIONS: The results provide evidence that, with few exceptions, satisfactory immunohistochemical staining is possible on plastic embedded bone marrow trephine biopsy specimens after microwave heating in citrate buffer. This, combined with the advantage of superior cellular morphology with semi-thin (1 micron) sections of plastic embedded material, make such embedding procedures the preferred method for the processing of bone marrow trephine biopsy specimens. PMID- 7490315 TI - Production and characterisation of an antimelanoma monoclonal antibody KBA.62 using a new melanoma cell line reactive on paraffin wax embedded sections. AB - AIMS--To generate new monoclonal antibodies directed against melanoma associated antigens using a new melanoma cell line, KAL. METHODS--The melanoma cell line was established in culture from a lymph node metastasis of malignant melanoma. Normal Balb/c mice were immunised with KAL cells. Splenocytes were used for fusion experiments using standard techniques. Hybridoma supernatants were tested for antibody binding activity using an indirect immunoperoxidase method on frozen sections from KAL tumour cells xenografted onto nude mice and human tonsils. KBA.62 was selected because of its reactivity with melanocytic proliferations on both frozen and paraffin wax sections. RESULTS--On immunoblotting, KBA.62 reacted with three bands of 140, 135 and 128 kD and two weak bands of 88 and 73 kD. In normal human tissues basal melanocytes in the epidermis did not react with this antibody and only occasional labelling of endothelial cells was noted. Of the human tumours, KBA.62 reacted strongly and uniformly with the majority of benign (21/21) and malignant (75/86) melanocytic proliferations. Staining was localised predominantly to the cell membrane with little or no cytoplasmic reactivity. Negative staining was observed in the majority of human non-melanocytic neoplasms, the exceptions being some carcinomas (11/89), particularly the well differentiated squamous cell type. This, however, was not thought to present a diagnostic problem. CONCLUSIONS--KBA.62 appears to be potentially useful in ascertaining the immunomorphological diagnosis of malignant melanoma in routinely processed paraffin wax sections. PMID- 7490317 TI - Circulating Ki67 positive lymphocytes in multiple myeloma and benign monoclonal gammopathy. AB - AIMS: To estimate the proportion and nature of the proliferating (Ki67+) circulating lymphocytes in a series of patients with multiple myeloma and monoclonal gammopathy of unknown significance (MGUS) and to correlate this with other clinical and laboratory parameters, using blood from healthy adults as a control. To investigate the extent to which the B and T lymphoid components are involved in progression and/or control of disease. METHODS: Blood lymphocytes from 15 patients with multiple myeloma, 10 patients with MGUS and 10 healthy adults were analysed using a sequential double immunoenzymatic staining technique. Antibodies directed against Ki67 were used to detect cells in cycle, CD3, CD4, and CD8 to identify T cells, HLA-Dr as a marker for B cells and activated T cells, and CD11b as a marker for natural killer cells. Polyclonal antibodies directed against the kappa and lambda immunoglobulin light chains were also used to detect B cells. RESULTS: The proportion of proliferating (Ki67+) lymphocytes was significantly higher in patients with multiple myeloma (6.8 +/- 2.6) and MGUS (3.5 +/- 1.1) compared with the normal controls (1.69 +/- 0.3); this was also true when multiple myeloma and MGUS cases were compared. In multiple myeloma and MGUS over 50% of the Ki67+ cells were activated T lymphocytes (CD3+/HLA-Dr+); a minority (11%) were non-clonal B lymphocytes. In contrast to controls (6.7 +/- 1.9), in patients with multiple myeloma and MGUS the proportion of proliferating T cells expressing CD8 (23.6 +/- 12.5 and 15.3 +/ 7.7, respectively) and CD11b (13 +/- 8.7 and 11.6 +/- 3.9, respectively) was higher. In multiple myeloma there was a positive correlation between the proportion of Ki67+ lymphocytes, beta-2-microglobulin concentrations and disease stage. CONCLUSIONS: Although the number of patients investigated is small, this study suggests that Ki67 expression in blood lymphocytes from patients with multiple myeloma may be a good prognostic indicator for aggressive disease and may help to distinguish multiple myeloma from MGUS. The activated proliferating T cells in these diseases may represent an immunological reaction against the tumour. PMID- 7490319 TI - Down-regulation of Epstein-Barr virus nuclear antigen 1 in Reed-Sternberg cells of Hodgkin's disease. AB - AIMS: To demonstrate Epstein-Barr virus (EBV) encoded nuclear antigen 1 (EBNA-1) gene expression in EBV associated disorders using a new monoclonal antibody (1H4 1) on routinely processed tissues. METHODS: The pressure cooker antigen retrieval method was used for the immunohistochemical demonstration of EBNA-1 gene expression in formalin fixed, EBV positive tissues from Hodgkin's disease, infectious mononucleosis, HIV associated non-Hodgkin's lymphomas, post-transplant lymphomas, and undifferentiated nasopharyngeal carcinoma (NPC). EBV encoded EBNA 2, latent membrane protein 1 (LMP-1) and BZLF-1 gene expression was also examined using commercially available monoclonal antibodies. RESULTS: Of the 34 EBER in situ hybridisation positive cases of Hodgkin's disease examined, none expressed EBNA-1 in the Reed-Sternberg cells. These cells were nevertheless strongly LMP-1 positive in all cases. Strong EBNA-1 staining was seen in all cases of EBER positive HIV associated non-Hodgkin's lymphoma (five of five), nasopharyngeal carcinoma (five of five), infectious mononucleosis (three of three), and post transplant lymphoma (one of one). These cases also expressed LMP-1, EBNA-2 and BZLF-1, but at differing levels. CONCLUSION: The pressure cooker antigen retrieval procedure is a sensitive and reliable adjunct to immunohistochemistry, especially with antibodies which are otherwise ineffective on routinely processed tissues. The EBNA-1 gene is not expressed at detectable levels in the malignant cells of Hodgkin's disease, but is consistently expressed in other EBV associated disorders. This finding has important implications for the role of EBNA-1 in the biology of EBV. PMID- 7490320 TI - Influence of local peritoneal involvement on pelvic recurrence and prognosis in rectal cancer. AB - AIMS: To evaluate the influence of involvement of the peritoneal surface by carcinoma of the rectum on local recurrence and prognosis. METHODS: Prospective analysis of pathological prognostic factors in 209 resections for rectal carcinoma between 1988 and 1993 with meticulous pathological technique particularly to assess the relation of tumour to the peritoneal surface. Comprehensive clinical follow up with cause of death established from all available sources of information (hospital and general practitioner data) with necropsies where necessary. Local recurrence was determined by accepted clinical, radiological and pathological criteria. RESULTS: Local peritoneal involvement was detected in 25.8% (54/209) of cases. It was more common in women and was associated with tumour differentiation, size and site, and lymph node involvement. Local peritoneal involvement showed considerable prognostic disadvantage in all cases and in curative cases alone. Multivariate analysis demonstrated independent prognostic disadvantage for all cases although this was lost in the curative group. With a 30 month median follow up time, comprehensive clinical surveillance detected 25 (12.0%) local recurrences. Thirteen (52%) palliative cases had shown spread to involve the mesorectal (deep, circumferential) resection margin. Of the 12 curative cases, six were upper rectal cancers with local peritoneal involvement suggesting that tumour seeding into the pelvic peritoneal cavity was the cause of local recurrence. Local recurrence of the six other rectal tumours was probably because of intraluminal seeding in two, involvement of the distal margin in one, extensive extramural venous involvement in two, and tumour spread to the bladder in one. CONCLUSIONS: Comprehensive pathological analysis of a resection specimen can identify cases with a high probability of local recurrence which may benefit from early adjuvant therapy. Involvement of the peritoneal surface is a common event in rectal cancer, has adverse prognostic influence and may be an important factor in local recurrence of upper rectal carcinoma. PMID- 7490321 TI - Malignant mesothelioma of the pleura: interobserver variability. AB - AIMS: To assess the consistency of histopathological diagnosis of pleural malignant mesothelioma by estimating interobserver agreement between five pathologists. METHODS: Eighty eight histological sets pertaining to original diagnoses of pleural malignant mesothelioma were reviewed separately by each pathologist. Diagnostic likelihood was graded as definite (A), probable (B), possible (C), improbable (D), and definitely not (E) malignant mesothelioma. The following indexes were estimated: observed proportion of agreement (Po), kappa statistics and proportion of agreement for "positive" (Ppos) and "negative" (Pneg) diagnoses. RESULTS: Sixty cases (68.2%) were rated by at least three reviewers as A or B and by none of the others as D or E. Five (5.7%) were rated by at least two reviewers as D or E and by none of the others as A or B. Nine (10.2%) showed a serious disagreement, rated A or B and D or E. Agreement for sets obtained at necropsy/surgery (median kappa w = 0.57) was similar to that for endoscopic material (median kappa w = 0.54). Agreement was poor on material obtained by needle biopsy (median kappa w = 0.21). The median value of Ppos varied between 0.94 (necropsy/surgery) and 0.67 (needle biopsy) and that of Pneg between 0.78 (necropsy/surgery) and 0.34 (unspecified biopsy). Interobserver agreement on histological typing was good overall (median kappa = 0.59). CONCLUSIONS: Of the original histological diagnoses, 70% were consistently reproduced through panel review. Most indexes of agreement between pathologists ranged from poor (needle biopsy) to moderate (necropsy/surgery). Agreement in confirming malignant mesothelioma was greater than that regarding exclusion of this diagnosis. Of the cases finally considered to have malignant mesothelioma, the reproducibility of histological typing was relatively satisfactory. PMID- 7490322 TI - Comparison of haemoglobin H inclusion bodies with embryonic zeta globin in screening for alpha thalassaemia. AB - AIMS: To compare the haemoglobin (Hb) H inclusion test with immunocytochemical detection of embryonic zeta chains in screening for alpha thalassaemia. METHODS: Blood samples from 115 patients with relevant clinical history and hypochromic microcytic indexes were screened using the HbH inclusion test and the Variant Hemoglobin Testing System (BioRad, Hercules, CA, USA). RESULTS: The HbH inclusion test was positive in 61 of 115 cases, three of whom had HbH disease confirmed by electrophoresis. The remaining 58 had alpha thalassaemia 1. All three HbH cases and 56 of 58 cases of alpha thalassaemia 1 expressed embryonic zeta chains, giving a specificity of 96.7%. Fifty four of 115 cases had a negative HbH inclusion test, of whom 50 had beta thalassaemia trait and three had iron deficiency. No diagnosis was reached for the remaining patient. CONCLUSION: The immunocytochemical test is as sensitive as the HbH inclusion test in screening for alpha thalassaemia. The presence of zeta chains is highly specific for alpha thalassaemia 1 incorporating the (--/SEA) deletion. The specificity and simplicity of the immunocytochemical test make it the test of choice in screening for alpha thalassaemia. PMID- 7490323 TI - Management of blood donors whose donations are repeatedly falsely positive by the HIV antibody screening test. AB - Since 1985, over 1,800,000 donations have been screened by the West Midlands Regional Blood Transfusion Service for antibody to HIV. Twelve regular donors gave three or more donations that were alternatingly positive and negative in the screening test, but not confirmed to be HIV positive by supplementary testing. Extensive investigation of six of these donors, including the polymerase chain reaction (PCR), failed to confirm HIV infection. The donors were reassured but, nevertheless, retired to comply with the guidelines of the National Blood Transfusion Service. These findings indicate that, for UK donors, ambiguous serological findings are unlikely to reflect HIV infection. On the rare occasions where serological results are particularly ambiguous, PCR testing of donors' blood may be helpful. PMID- 7490324 TI - Osteolytic bone lesions in a patient with idiopathic myelofibrosis and bronchial carcinoma. AB - A 59 year old man with longstanding myelofibrosis and previous splenectomy was incidently found to have a large lytic lesion in his left femur which required operative fixation. He had undergone right upper lobectomy for squamous carcinoma of the bronchus five years earlier. Histological analysis of bone reamings showed no evidence of metastatic carcinoma. Osteosclerosis is frequently noted in patients with myelofibrosis but osteolytic lesions are uncommon and may be confused with metastatic malignancy. PMID- 7490325 TI - Combined goblet cell carcinoid and mucinous cystadenoma of the appendix. AB - Two cases of combined goblet cell carcinoid and mucinous cystadenoma occurring in the appendix are reported. The histogenesis of the goblet cell carcinoid remains one of its most controversial aspects and the occurrence of both of these relatively uncommon tumours in the same organ may lend support to the unitary stem cell hypothesis on the origin of this tumour. Alternatively, this occurrence may represent an example of the adenoma/carcinoma sequence. PMID- 7490326 TI - Hodgkin's disease in a patient with common variable immunodeficiency. AB - A 61 year old man with long standing common variable immunodeficiency presented with pyrexia, anaemia and leucopenia. A diagnoses of Hodgkin's disease of the bone marrow was made. The typical histopathological and immunophenotypic appearances were clearly distinct from those of T cell lymphoma with Reed Sternberg-like cells which, in contrast to Hodgkin's disease, is a known complication of common variable immunodeficiency. Complete clinical and histological remission was achieved with combination chemotherapy. The latter was complicated by severe myelosuppression, unusually severe erosive mucositis and viral retinitis. PMID- 7490327 TI - Immunohistochemical analysis of T cell proliferation in normal tonsil and B cell lymphoma. AB - A double immunohistochemical technique, incorporating MIB1 and CD3, was used to identify proliferating T cells in paraffin wax sections of normal tonsil and B cell lymphomas. The number of double stained T cells as a percentage of the total T cells was then determined. In normal tonsil and follicular lymphoma the follicle centre and T cell zones were counted independently. In normal tonsil very few T cells in the follicle centre expressed MIB1. Proliferating T cells were concentrated in the T cell zones. The same pattern was observed in follicular lymphoma. In contrast, the percentage of T cells expressing MIB1 was higher in mucosa associated lymphoid tissue type lymphoma and lymphocytic lymphoma, suggesting that T cell activation occurs in these tumours. The highest percentage of MIB1 positive T cells was observed in high grade lymphoma. This suggests that transformation to high grade lymphoma is associated with an increase in T cell activation. PMID- 7490328 TI - A "quickscore" method for immunohistochemical semiquantitation: validation for oestrogen receptor in breast carcinomas. AB - Immunohistochemistry is increasingly used in the assessment of markers for breast cancer prognosis. Semiquantitation is frequently desirable but, other than by the use of image analysis, the approaches currently in use are cumbersome. The most common method used is the H-score which takes into consideration the staining intensity in conjunction with the percentage of cells staining positively in breast carcinoma tissue. A "quickscore" has been developed which dispenses with the need to count individual cells. The quantitative biochemical Abbott enzyme immunoassay (EIA) and the Dako immunohistochemical assay (IHA) incorporating a semiquantitative H-score, have been used as standards against which the IHA quickscore for the semiquantitation of oestrogen receptor expression was tested. A good correlation was found between the quickscore and the EIA, which was as good as that between the H-score and EIA. The quickscore is a valid approach and there is no advantage in using the more rigorous H-score. A positive cut off quickscore of > or = 3 has been suggested. PMID- 7490329 TI - Granulomatous bone marrow inflammation during treatment of chronic myeloid leukaemia with interferon alpha-2b. AB - A patient with chronic myeloid leukaemia developed bone marrow granulomas during treatment with interferon alpha-2b. Some granulomas had necrotic centres and giant cells and there was marked eosinophilia surrounding them. The granulomas disappeared when the interferon treatment was discontinued. Mycobacteriosis was ruled out. The most likely explanation for the granuloma formation was drug hypersensitivity. PMID- 7490330 TI - Hepatitis C virus replication in hepatocellular carcinoma. AB - Hepatitis C virus (HCV) replication is reported in both tumour and non-tumour tissue in a case of hepatocellular carcinoma. Viral replication was established by showing the presence of minus strand HCV RNA by PCR amplification, after excluding residual reverse transcriptase activity of Taq polymerase. No minus strand was found in serum derived virion RNA. PCR amplified products from both tumour and non-tumour parenchyma were sequenced in the 5' non-coding region and shown to be identical. The genotype of this Indonesian patient was found to be 1b (or II), the most prevalent type in the Far East. PMID- 7490331 TI - Seasonal variation in the necropsy incidence of pulmonary thromboembolism. PMID- 7490332 TI - Should superheating be used for immunohistochemistry? PMID- 7490333 TI - CD34 immunoperoxidase staining for the diagnosis of myelodysplastic syndromes and chronic myeloid leukaemia. PMID- 7490334 TI - Mycoplasma mycoides subspecies mycoides (small colony variant): the agent of contagious bovine pleuropneumonia and member of the "Mycoplasma mycoides cluster". PMID- 7490335 TI - Pathology of equine respiratory disease occurring in association with transport. AB - Eight young thoroughbred horses, taken 1858 km by road (travelling time, 41 h), were examined to assess the pathological nature of respiratory disease associated with transport. Three of the horses showed clinical abnormalities including pyrexia, coughing, leucocytosis and neutrophilia after the first 20 h of transportation. Endoscopical examination of the trachea revealed exacerbation of airway inflammation as a result of transport in two of the three affected horses. A consistent finding in the affected horses was focal serous neutrophilic pneumonia affecting the cranio-ventral portion of the caudal lung lobe with a propensity to affect the right lung. Streptococcus equi subspecies zooepidemicus was isolated from the pneumonic areas, in which corresponding bacterial antigens were identified immunohistochemically. Viral cultures from the pneumonic lesions proved negative for respiratory viruses. It is suggested that transport predisposes the upper respiratory tract and the lower airways to invasion by the bacterium, with episodic pyrexia and acute pneumonia. PMID- 7490336 TI - Assessment of the nucleolar organizer regions by automated image analysis in benign and malignant colonic tumours and adjacent tissues in rats. AB - An automated computer image analysis technique was used to study the morphological parameters of argyrophilic nucleolar organizer regions (AgNORs) in chemically induced rat colonic tumours of different grades. Different nuclear parameters were of different diagnostic value. For distinguishing tumorous tissue from normal tissue all the parameters studied were capable of serving as diagnostic markers. Malignant processes could, however, be more reliably detected by means of the area of the nucleus, nuclear shape factor, area of AgNOR and ratio of AgNOR area to nuclear area. In macroscopically normal tissue adjacent to a tumour, the values of all the AgNOR parameters studied were similar to those in tumorous tissues. It can be concluded that the initial stages in tumorigenesis are accompanied by changes in all of the nucleus and AgNOR parameters, but as malignancy develops, only some of these parameters continue to change. Close correlations between nuclear parameters in malignant tissue suggest that for diagnostic purposes only nuclear and AgNOR areas should be used. PMID- 7490337 TI - Pathogenesis of foot-and-mouth disease in swine, studied by in-situ hybridization. AB - Eight 7-month-old pigs were inoculated intradermally with 10(3) plaque-forming units of foot-and-mouth disease virus, type O, and killed 24, 48, 72, or 96 h later. Numerous tissues from each animal were collected and examined histopathologically and by in-situ hybridization to determine the presence of virus and its correlation with lesion development. The probe for in-situ hybridization was a biotinylated 500-base negative-sense transcription product corresponding to a portion of the gene encoding polymerase. With this technique, virus was shown to be widely disseminated in all epidermal tissues, regardless of histologically apparent cellular disruption. PMID- 7490338 TI - An immunohistochemical investigation of porcine epidemic diarrhoea. AB - A sudden outbreak of epidemic diarrhoea of piglets occurred in Japan, the principal features being watery diarrhoea, dehydration and high mortality in newborn animals. The microscopical lesions were villous atrophy in the small intestine, the villous enterocytes being vacuolated and cuboidal in shape. The villus-crypt ratio was severely reduced, varying from 1:1 to 3:1. Transmission electron microscopy showed numerous coronaviruses within the cytoplasm of enterocytes and among microvilli. Specific antigens of porcine epidemic diarrhoea (PED) virus were detected in the cytoplasm of enterocytes by the streptavidin biotin (SAB) technique. Infected cells, which were most abundant in the villous epithelia of the jejunum and ileum, were present in small numbers in the large intestine, the crypt epithelia, the lamina propria and Peyer's patches. The study suggests that the SAB technique is useful for the diagnosis of PED. PMID- 7490339 TI - Silver-stained nucleolar (Ag-NOR) cluster size in feline mammary carcinomas: lack of correlation with histological appearance, mitotic activity, tumour stage, and degree of nuclear atypia. AB - The relationship between Ag-NOR cluster size, mitotic index, tumour stage, degree of nuclear atypia and histological appearance was evaluated in a series of feline mammary carcinomas. The mean Ag-NOR cluster area per cell in these tumours ranged from 1.86 to 6.25 microns 2 and was not related to any other parameter studied. This suggests that Ag-NOR cluster size measurement is of poor diagnostic and prognostic value for feline mammary carcinomas. PMID- 7490340 TI - Histopathology of C57BL/6 mice inoculated orally with Mycobacterium paratuberculosis. AB - The susceptibility of C57BL/6 mice to oral inoculation with Myobacterium paratuberculosis was evaluated histopathologically. Granulomatous lesions containing acid-fast bacteria developed in the mesenteric lymph nodes in over 50% of the mice by 11 months after inoculation. The results suggest that C57BL/6 mice may be useful for studying infection, pathogenesis, and other aspects of paratuberculosis. PMID- 7490341 TI - Secondary retinoschisis in a dog. AB - Retinoschisis, an extreme form of cystic degeneration of the retina, was identified as a diffuse, bilateral microscopical change in an 8-month-old, male, English springer spaniel dog with a clinical history of blindness, retinal detachment and glaucoma. The absence of any material in the cystic spaces and the spectrum of intraocular degenerative changes indicated that the retinal change was a secondary retinoschisis, probably due to retinal detachment. Separation of retinal layers without disruption of the blood supply probably plays a part in the aetiology of retinoschisis. The pathogenesis and natural history of intraretinal cystic changes and retinoschisis in animals are poorly understood. PMID- 7490342 TI - Rhodococcus equi-associated necrotizing lymphadenitis in a llama. AB - A case of Rhodococcus equi-associated necrotizing lymphadenitis in a 2-year-old male llama is described. Caseous necrosis, resembling macroscopically that seen in ovine caseous lymphadenitis, was observed diffusely in the tracheobronchial and mediastinal lymph nodes, and in an extensive lesion in the lungs. Necrosis was present to a lesser extent in the spleen and hepatic and gastric lymph nodes. Numerous bacteria-laden macrophages were present around the necrotic areas. The findings suggest that, as in cattle and pigs, the primary targets of R. equi infection in the llama are the lymphoid organs. PMID- 7490343 TI - Kinetics of interferon gamma production in vivo during infection with the S48 vaccine strain of Toxoplasma gondii. AB - The kinetics of interferon gamma (IFN gamma) production in vivo was examined in sheep during a primary and secondary infection with the sheep vaccine strain (S48) of Toxoplasma gondii. Efferent lymph plasma from a node draining the site of inoculation was tested for anti-viral activity which could be neutralized with monoclonal antibodies against IFN gamma. Within 2 to 5 days of primary infection IFN gamma was detected in each of five sheep and persisted for 6 to 9 days. Accelerated production of IFN gamma occurred after secondary infection, the cytokine being detected in the first 24 h, and persisting in lymph for a further 4-5 days. From day 6 onwards after primary infection, efferent lymph cells produced IFN gamma when stimulated in vitro with a crude T. gondii antigen. These results show that IFN gamma is induced in sheep after infection with the S48 strain of T. gondii. PMID- 7490344 TI - Interferon gamma inhibits the intracellular multiplication of Neospora caninum, as shown by incorporation of 3H uracil. AB - An assay was developed to quantify the growth of two different isolates of the protozoon Neospora caninum within ovine fibroblast cells in vitro by differential uptake of 3H uracil. The NC-1 isolate of N. caninum multiplied more quickly in culture than the NC Liverpool isolate, as reflected by increased incorporation of isotope by the former over a shorter period of time. After the parasites had left the ruptured host cells, there was very little incorporation of isotope. This suggested that multiplication occurred within and not outside the cells. Treatment of the cells with ovine recombinant interferon gamma for 24 h before infection significantly inhibited intracellular multiplication of the parasite. PMID- 7490346 TI - Atopic dermatitis: is it an allergic disease? AB - For many years controversy has surrounded the relation between allergy and atopic dermatitis. We critically review the evidence for the contribution of allergy, or IgE-mediated hypersensitivity reactions, to the pathogenesis of this disease. We conclude that, at present, there is scant evidence that allergy is central to the development of atopic dermatitis, although it may be an aggravating factor in a few patients. Hence there is little rationale for the routine use of allergy testing or dietary and environmental manipulation in the management of this disease. PMID- 7490347 TI - Dog, cat, and human bites: a review. AB - It is estimated that half of all Americans will be bitten by an animal or another human being during their lifetimes. The vast majority of the estimated 2 million annual mammalian bite wounds are minor, and the victims never seek medical attention. Nonetheless, bite wounds account for approximately 1% of all emergency department visits and more than $30 million in annual health care costs. Infection is the most common bite-associated complication; the relative risk is determined by the species of the inflicting animal, bite location, host factors, and local wound care. Most infections caused by mammalian bites are polymicrobial, with mixed aerobic and anaerobic species. The clinical presentation and appropriate treatment of infected bite wounds vary according to the causative organisms. Human bite wounds have long had a bad reputation for severe infection and frequent complication. However, recent data demonstrate that human bites occurring anywhere other than the hand present no more of a risk for infection than any other type of mammalian bite. The increased incidence of serious infections and complications associated with human bites to the hand warrants their consideration and management in three different categories: occlusional/simple, clenched fist injuries, and occlusional bites to the hand. This article reviews dogs, cat, and human bite wounds, risk factors for complications, evaluation components, bacteriology, antimicrobial susceptibility patterns, and recommended treatments. Epidemiology, clinical presentation, and treatment of infections caused by Pasteurella multocida, Capnocytophaga canimorsus, Eikenella corrodens, and rhabdovirus (rabies only) receive particular emphasis. PMID- 7490348 TI - Dermatology in the cinema. AB - The depiction of skin disease in the cinema can be divided into three categories. These include skin findings on actors independent of the roles they portray, cutaneous disease used as a representation of evil, and skin disease represented realistically and sympathetically. Examples from a wide range of films are given, and implications for dermatologists and their patients are discussed. PMID- 7490349 TI - Surgical pearl: tips for increasing efficiency during hair transplantation--the sixteen-bladed knife. PMID- 7490345 TI - Benign melanocytic lesions: risk markers or precursors of cutaneous melanoma? AB - BACKGROUND: The role of benign melanocytic lesions as precursors and not only as risk markers for the development of cutaneous melanoma is controversial. OBJECTIVE: The purpose of the study was to assess the frequency of the histologic association of benign melanocytic lesions with cutaneous melanoma of a maximum thickness of 1.00 mm. The possibility that the spatial association of benign lesions with melanoma may be coincidental was also investigated. METHODS: The study subjects representing 289 cases of cutaneous melanoma of maximum thickness 1.00 mm (or less) were examined histologically for the presence of an associated benign melanocytic lesion(s), including lentiginous melanocytic proliferation; junctional, compound, or intradermal nevus; dysplastic nevus; and congenital nevus contiguous with or adjacent to the melanoma. The effects of age, tumor thickness, level of invasion, histologic type, and anatomic site on the association of benign melanocytic lesions with melanoma were assessed. In the control subjects 40 basal cell carcinomas and 38 compound nevi (not dysplastic) randomly chosen and matched for age (+/- 1 year) and site (head/neck, trunk, upper and lower limbs) with a melanoma case were examined to assess the proportion of these cases associated with benign lesions compared with the matched melanoma cases. RESULTS: A nevus was associated with melanoma in 51% of cases (n = 147). Of these, 82 (56%) were dysplastic nevi, 61 (41%) were common acquired nevi, and 4 (3%) were congenital nevi. Lentiginous melanocytic proliferation was present in the epidermis adjacent to 219 melanomas (75%) and in 44% of these cases (n = 97) a coexisting nevus was also present. CONCLUSION: The results of this study lend further support to the concept of common acquired nevi and dysplastic nevi as precursors of cutaneous melanoma. In addition, lesions diagnosed clinically as simple lentigo and solar lentigo may be important as potential precursors of melanoma, particularly in the elderly. PMID- 7490350 TI - Cafe-au-lait macule in type V skin: successful treatment with a 510 nm pulsed dye laser. PMID- 7490351 TI - Granulomatous slack skin: a possible precursor of Hodgkin's disease. PMID- 7490352 TI - Human papillomavirus DNA in nonanogenital keratoacanthoma and squamous cell carcinoma of patients with HIV infection. PMID- 7490353 TI - Destructive facial porokeratosis. PMID- 7490354 TI - Dermatitis herpetiformis associated with lichen planopilaris. PMID- 7490356 TI - Indications and complications of medicinal leech therapy. PMID- 7490355 TI - Acquired multiple blue nevi on an extremity. PMID- 7490357 TI - State and territorial regulation of tattooing in the United States. PMID- 7490358 TI - Antiandrogens for penile surgery. PMID- 7490359 TI - Lichen sclerosus. PMID- 7490360 TI - Sezary syndrome in association with silicone breast implant. PMID- 7490361 TI - The minigrafting test for vitiligo: detection of stable lesions for melanocyte transplantation. PMID- 7490362 TI - Lentigo maligna and lentigo maligna melanoma. AB - Lentigo maligna (LM) is a pigmented lesion that occurs on the sun-exposed skin, particularly the head and neck areas, of an older patient. The lesion increases in size and at some point, often many years after its onset, may become lentigo maligna melanoma (LMM). For this reason, most authors consider LM a form of melanoma in situ. Treatment includes surgical or destructive modalities; the preferred form of therapy is surgical removal. Histopathologic features include a proliferation of atypical melanocytes along the basal layer of the epidermis and adnexal structures. This article discusses the clinical, histopathologic, and epidemiologic features of LM. The prognosis and treatment of LM are reviewed. Although the lifetime risk of the development of LMM is unclear, LMM is discussed briefly. PMID- 7490363 TI - The effects of chronic sunscreen use on the histologic changes of dermatoheliosis. AB - BACKGROUND: Sunscreen application to the skin of hairless mice is effective in reversing the histologic changes associated with photoaging (solar elastosis, epidermal thickening, collagen depletion, glycosaminoglycan deposition). These reparative processes have not been studied in human beings. OBJECTIVE: The aim of this study was to evaluate histologically the effects of daily application of a UVA/UVB sunscreen versus placebo. METHODS: We examined 46 patients who were given either sunscreen or vehicle and asked to apply it daily for 24 months. Punch biopsy specimens were obtained from preauricular skin at 0, 12, and 24 months. Each specimen was examined for epidermal thickening and organization and dermal inflammatory infiltrate by light microscopy. Computer-generated analysis of tissue sections was used to evaluate solar elastosis. RESULTS: A significant difference in solar elastosis was found between the treatment groups; however, the other features remained largely unchanged. CONCLUSION: The dermal changes of photoaging may be affected differently than epidermal changes when UV radiation exposure is diminished. UVA and UVB may contribute diversely to these cutaneous changes. Computer-generated evaluation of dermatoheliosis may be more accurate than visual inspection. PMID- 7490364 TI - Effects of cyclosporine on cytokines and cytokine receptors in psoriasis. AB - BACKGROUND: Oral cyclosporine is effective in the treatment of recalcitrant psoriasis. However, the precise mechanism(s) are not fully understood. A possible mode of action may be via down-modulation of proinflammatory cytokines that are increased in psoriatic lesions. OBJECTIVE: This study was designed to monitor the effects of cyclosporine treatment on the expression of cytokines, cytokine receptors, and other markers of inflammation in psoriatic skin. METHODS: Ten patients with recalcitrant psoriasis were treated with cyclosporine. The in vivo effects of cyclosporine on cytokines and their receptors were studied by the use of cryostat-cut sections and a panel of antibodies. The in vitro effects were studied with flow cytometry of epidermal cell suspensions prepared from psoriatic lesions and control skin. RESULTS: Clinical improvement was noted in all patients after 2 weeks of cyclosporine treatment. The expression of interleukin-1 beta, interleukin-8, CD25(IL-2R), CD36 and E-selectin were significantly decreased, whereas the number of tumor necrosis factor-receptor-positive epidermal cells was significantly increased in psoriatic lesions. CONCLUSION: Clinical improvement of psoriasis with cyclosporine treatment is accompanied by down modulation of proinflammatory epidermal cytokines and decreased dermal inflammation. Thus besides suppressing cytokine production by the inflammatory infiltrate, the beneficial effect of cyclosporine in psoriasis also depends on the inhibition of the epidermal cytokine network. PMID- 7490365 TI - Uremic small-artery disease with medial calcification and intimal hyperplasia (so called calciphylaxis): a complication of chronic renal failure and benefit from parathyroidectomy. AB - BACKGROUND: Uremic small-artery disease with medial calcification and intimal hyperplasia can lead to life-threatening skin necrosis or acral gangrene. It is a distinct complication of chronic renal failure that must be differentiated from soft-tissue calcification. An increased calcium-phosphate product and secondary hyperparathyroidism are the main underlying conditions. The benefit of parathyroidectomy is controversial. OBJECTIVE: This article is based on a literature search to determine prognostic factors and, in particular, the benefit of parathyroidectomy. METHODS: The literature on uremic small-artery disease (so called calciphylaxis) was reviewed (full data set: 104 cases, including five of our own). The therapeutic benefit of parathyroidectomy and the relation between prognostic predictors (localization, dialysis, and transplant) and outcome were analyzed. The relation between diabetes and acral gangrene was also examined. Further epidemiologic data on the reviewed group of patients were established. RESULTS: Thirty-eight of 58 patients who underwent parathyroidectomy survived compared with 13 of 37 patients who did not undergo parathyroidectomy (p = 0.007, n = 95). Forty of 53 patients with distal localization of necrosis survived compared with 11 of 42 patients with proximal pattern (p < 0.00001; n = 95). Dialysis and kidney transplantation followed by immunosuppression showed no relation to disease outcome. No association was found between diabetes and acral gangrene (p = 0.50). CONCLUSION: Uremic small-artery disease is a distinct complication of chronic renal failure. Its recognition and early diagnosis should allow more effective treatment. In our retrospective study parathyroidectomy was significantly related to survival. Only a randomized, controlled, prospective trial (parathyroidectomy vs conservative treatment of secondary hyperparathyroidism) can establish the value of parathyroidectomy in uremic small artery disease. PMID- 7490366 TI - Reactions to other corticosteroids in patients with positive patch test reactions to budesonide. AB - BACKGROUND: Corticosteroid cross-reactions have been classified into four well defined groups. A previous study of patch test reactions to other corticosteroids in patients allergic to hydrocortisone failed to conform to these groups. It was suggested that substitution at the C6 and C9 positions of the corticosteroid was the most important determinant of a further reaction. OBJECTIVE: Our aim was to analyze multiple positive patch tests to corticosteroids in patients sensitized to budesonide to confirm our earlier findings. METHODS: Forty-six patients with positive patch tests to budesonide were patch-tested to 17 other topical corticosteroids. The results were examined by a generalized linear model and a chi-square test. RESULTS: Substitution of the corticosteroid at the C6 and C9 positions significantly reduced the number of reactions. A different substitution at the C16 and C17 positions was less important, and that at the C21 position was of no significance. CONCLUSIONS: Patients sensitized to budesonide are most likely to react to other non-C6 and non-C9 substituted corticosteroids. PMID- 7490367 TI - Improvement of skin barrier function during treatment of atopic dermatitis. AB - BACKGROUND: Active dermatitis causes a disturbance in skin barrier function. This can be evaluated by the measurement of transepidermal water loss (TEWL) and percutaneous absorption of hydrocortisone. OBJECTIVE: The study objective was to evaluate changes in skin barrier function during treatment of atopic dermatitis. METHODS: Nine patients with widespread atopic dermatitis were studied longitudinally by measuring the severity of the dermatitis and TEWL at intervals of 1 to 3 days. Percutaneous absorption of hydrocortisone was measured at entry and during treatment. RESULTS: At entry, both TEWL and percutaneous absorption of hydrocortisone were elevated. Four to six days later, a significant decline was observed in both variables, indicating rapid improvement in skin barrier function. Individual changes in TEWL correlated with the changes in the systemic absorption of hydrocortisone. CONCLUSION: TEWL reflects changes in the systemic absorption of topical hydrocortisone during treatment of atopic dermatitis. PMID- 7490368 TI - Serum tissue inhibitor of metalloproteinases in patients with systemic sclerosis. AB - BACKGROUND: One of the suggested contributory factors to the development of dermal fibrosis is a decrease in collagenase activity, which may be related to levels of serum tissue inhibitors of metalloproteinase-1 (TIMP-1). OBJECTIVE: The aim of this study was to determine the clinical significance of serum TIMP-1 levels in systemic sclerosis (SSc). METHODS: We measured serum TIMP-1 concentration in 62 patients with SSc, 11 patients with systemic lupus erythematosus, 14 patients with rheumatoid arthritis, and 22 members of a normal control group. The clinical features of the patients with SSc and elevated TIMP levels were examined. RESULTS: The mean TIMP-1 level in the patients with SSc was significantly higher than that in the members of the control group or the patients with systemic lupus erythematosus or rheumatoid arthritis. In 44% of the patients with SSc the serum TIMP-1 level was elevated. The mean serum TIMP-1 level in patients with diffuse cutaneous SSc (dSSc) was significantly higher than that in those with limited cutaneous SSc. The patients with dSSc and elevated serum TIMP-1 levels showed a significantly greater incidence of lung fibrosis and anti-topoisomerase I antibody than those with normal serum TIMP-1 levels. The TIMP-1 level and diffusing capacity for carbon monoxide in the patients with SSc were negatively correlated. Increased mitogenic activity on dermal fibroblasts caused by serum from patients with dSSc was partially blocked by anti-TIMP-1 IgG. CONCLUSIONS: These findings suggest that serum TIMP-1 level is a useful indicator of disease activity in patients with SSc and that TIMP is involved in the pathogenesis of SSc. PMID- 7490369 TI - Tumor of the follicular infundibulum: a clinicopathologic study. AB - BACKGROUND: Tumor of the follicular infundibulum (TFI) is a rare benign adnexal tumor that has characteristic histopathologic features. OBJECTIVE: Our purpose was to describe the clinical and pathologic features of 12 patients with TFI. METHODS: Of 121,500 cutaneous biopsy specimens recorded between 1981 and 1993, all TFIs were identified and examined by conventional microscopy. The clinical and histologic findings were compared with those of previously published cases. RESULTS: Of the 12 patients, seven had a solitary tumor, two had eruptive TFI involving the face, and three TFIs were incidentally discovered in association with nevus sebaceus or fibroma. TFI-like changes were also observed in the wall of a hybrid cyst. The most discriminating features of TFI were the horizontal platelike organization of the tumor and the dense elastic network beneath the tumor. Of the different forms of TFI, only eruptive tumors can be clinically identified because they are small hypopigmented macules. CONCLUSION: We classify TFIs as (1) solitary tumors, (2) eruptive tumors, (3) TFI associated with other lesions of Cowden's disease, (4) TFI associated with a single tumor such as nevus sebaceus, and (5) TFI-like epidermal changes. PMID- 7490370 TI - Epidermal expression of 65 and 72 kd heat shock proteins in psoriasis and AIDS associated psoriasiform dermatitis. AB - BACKGROUND: Psoriasiform dermatitis is common in patients with AIDS. The expression of heat shock proteins by keratinocytes has been postulated to be a significant factor in the physiopathology of psoriasis and might be subject to modulation in HIV-infected patients. OBJECTIVE: We sought to evaluate the epidermal expression of 65 and 72 kd heat shock proteins (HSPs) in lesions of AIDS-associated psoriasiform dermatitis (AIDS-PD) and compare it with that in psoriasis vulgaris and seborrheic dermatitis in patients not infected with HIV. METHODS: Sections from paraffin-embedded blocks of biopsy specimens of AIDS-PD (eight cases), psoriasis vulgaris (eight cases), seborrheic dermatitis (four cases), and normal skin (four cases) in non-HIV-infected patients were immunohistochemically stained by the avidin-biotin-peroxidase method and two monoclonal antibodies directed against the major 65 kd HSP antigen (HSP65) and against 70/72 kd HSP. The intensity, distribution, and cellular pattern of the epidermal stain were graded and assessed blindly. RESULTS: The epidermal expression of HSP65 in biopsy specimens from AIDS-PD lesions was irregular, with less intensity and less tendency to perinuclear arrangement than in psoriasis or seborrheic dermatitis not associated with AIDS. The expression of HSP72 was also less intense and more uniform in AIDS-PD. CONCLUSION: The altered interplay of T cells and keratinocytes in a situation of immune derangement such as AIDS might account for the differences observed in the expression of HSP65 and HSP72 by keratinocytes in psoriasis and AIDS-PD. PMID- 7490371 TI - Repigmentation in vitiligo vulgaris by autologous minigrafting: results in nineteen patients. AB - BACKGROUND: Minigrafting is a successful therapy for localized vitiligo but has never been reported for vitiligo vulgaris. OBJECTIVE: Our purpose was to evaluate the efficacy of minigrafting in vitiligo vulgaris. METHODS: In 59 patients with stable vitiligo vulgaris, a minigraft test was done by implanting two minigrafts in the lesion to be grafted. Patients were selected for grafting when spread of pigment was observed within 3 months. The rate of repigmentation was evaluated by digital image analysis. RESULTS: Twenty-three patients (36 lesions), of 24 with a positive minigraft test, were grafted. The results of 19 patients were analyzed, showing 80% to 99% repigmentation in 14 lesions, 50% to 80% repigmentation in 10 lesions, and zero to 50% repigmentation in 12 lesions. Time of observation varied from 3 to 12 months after grafting. Best results were observed after 9 to 12 months. In all patients with a positive Koebner phenomenon depigmentation of the minigrafts developed. CONCLUSION: Autologous minigrafting is an effective therapy for stable vitiligo vulgaris in a selected group of patients. PMID- 7490374 TI - An overview of caries risk assessment, and its potential utility. AB - The clinical findings that dentists use to estimate the future caries activity of patients are imperfect. Almost invariably there are protective factors in patients who go on to develop clinical caries and there are risk factors that persist in patients who do not develop clinical caries. Therefore, the clinical decision process is characterized by a level of uncertainty concerning the selection of patient management strategies for caries. To the degree that information relating to caries risk assessment can reduce the amount of uncertainty associated with decisions for clinical patient care in the present, that information has utility. Dentists face many treatment decisions daily. The integration of a mechanistic understanding of caries and use of clinical data may yield useful information that will improve clinical patient management. Certain factors affect the potential utility of caries risk screening from the clinician's perspective. These can be summarized in terms of the costs and benefits of the procedures as well as the availability and appeal of alternative procedures. Factored in with these considerations are the investment history one has made toward a given procedure and the propensity for changing one's behavior. These considerations are not easily addressed at present since costs and benefits for different therapeutic approaches have been developed for only a few dental conditions. We have put forth a framework that emphasizes the clinician as the key component in the assessment of caries risk. We argue that integration of the clinician's focus with a mechanistic understanding of caries and a probabilistic, empirical appreciation for caries activity will be beneficial for patient management. These arguments remain untested and should be challenged in a research setting. We need to establish whether information derived from risk assessment procedures leads to improved clinical outcomes. We need to have a better understanding of how clinicians process clinically available risk information. We need to determine what additional risk assessment information will improve the clinician's ability to identify high-risk patients. We also need to establish whether there are differences in the usefulness of additional risk information depending on the level of experience of the clinician. PMID- 7490373 TI - Developing clinical teaching methods for caries risk assessment: introduction to the topic and its history. PMID- 7490372 TI - UVB phototherapy for pityriasis rosea: a bilateral comparison study. AB - BACKGROUND: Pityriasis rosea is a common self-limiting disease. UV light has been mentioned as helpful, but only a few studies substantiate this possibility. OBJECTIVE: Our purpose was to evaluate the clinical efficacy of UVB phototherapy in pityriasis rosea and the course of the disease after treatment. METHODS: Seventeen patients with extensive pityriasis rosea were treated with unilateral UVB phototherapy in a bilateral comparison study. One joule of UVA was used as a "placebo" on the untreated side. The treatments were given five times per week for 2 weeks. The rate of clearing was monitored by estimation of the severity score. RESULTS: Ten daily erythemogenic exposures of UVB resulted in substantially decreased severity of disease, greater than that on the untreated side in 15 of 17 patients. The overall reduction of severity score showed a significant difference after the third treatment; the UVB irradiation was superior. However, during the follow-up period the two sides were indistinguishable in regard to severity score (p = 0.6784). No significant difference in pruritus was noted between the two sides (p > 0.1638). The duration of disease did not relate to the success of UVB phototherapy. CONCLUSION: During the treatment period UVB phototherapy resulted in decreased severity of disease. However, the itching and the course of the disease were unchanged. PMID- 7490375 TI - Prediction of future high caries increments for children in a school dental service and in private practice. PMID- 7490376 TI - Caries risk assessment for determination of focus and intensity of prevention in a dental school clinic. PMID- 7490377 TI - Caries risk assessment used as a determinant for caries management and prevention. PMID- 7490378 TI - Assessment of caries risk factors and preventive practices. PMID- 7490379 TI - Caries risk assessment and prevention for adults. PMID- 7490380 TI - Critique of evolving methods for caries risk assessment. PMID- 7490381 TI - Applicant analysis 1994. PMID- 7490382 TI - An epidemiologic study of fall-related fractures among institutionalized older people. AB - OBJECTIVES: The purpose of this study was to assess the incidence of fall-related fractures, and the circumstances surrounding these events, during a 5-year period among all residents of a long-term care facility. PARTICIPANTS: The study group was composed of residents with radiologically confirmed fractures that were the direct result of a fall occurring between the years 1988 and 1992. This group was 82% female and had a mean age of 89 +/- 6 years. DESIGN: The study was designed as a 5-year retrospective cohort study in a long-term care institution where annual incidence rates of fracture were assessed. RESULTS: There were 296 fall related fractures during the 5-year period. Annual incidence rates remained fairly constant (72, 86, 84, 70, and 94 per 1000 person-years, respectively) despite a 54% decline in the use of physical restraints during the 5-year period. Hip fractures comprised almost 50% of all fractures. Based on incident reports of these fractures, 42% occurred during the day, 55% took place in the bedroom or adjoining bathroom. 67% occurred while the resident was ambulating, and a wet floor was in evidence in 16% of the incidents. CONCLUSION: Fracture incidence in this long-term care facility has remained stable despite reduced restraint use. The incident reporting system may contribute valuable information to the identification of factors that should receive further attention in studies of risk factors for fracture in the long-term care setting. PMID- 7490383 TI - Higher respiratory infection rates on an Alzheimer's special care unit and successful intervention. AB - OBJECTIVE: To determine the rates of various types of infections on an Alzheimer's special care unit (ASCU) compared with the rates found on traditional nursing home units. Because patients on the ASCUs are allowed to wander throughout the unit and typically come into contact with each other more frequently, we hypothesized that the rate of communicable infections such as upper respiratory infections are significantly higher than on other units where patients are more easily isolated when sick. METHODS: A 4-year retrospective case control study, 1990-1993. SETTING: A metropolitan long-term care skilled nursing facility. Three floors are traditional nursing care units (123 beds), and one floor is the ASCU (41 beds). PRIMARY OUTCOME: Annual nosocomial infection rates per 10,000 patient days were measured for six types of infection during the 1990 1993 study period. Data were segregated by location of infection, either the traditional nursing units or the ASCU. In 1992, patients on the Alzheimer's unit were placed in smaller activity groups, and an education program for the control of infectious agents was provided to the unit's staff. RESULTS: The relative order of prevalence for the different infection types remained constant during the 4 years. The most common type of infection for all 4 years of the study period was urinary tract infection (UTI), followed by upper respiratory infection (URI), Lower respiratory tract infection (LRI), cutaneous infection, gastrointestinal (GI) infection, and eye infection. Of these various infections, only URI rates remained consistently higher on the ASCU versus the traditional nursing unit over the 4-year study period (in years 1990, 1991, and 1993; these differences were statistically significant, P < .05). In 1992, the year in which nursing interventions to curb the relatively high rates of URI on the ASCU took place, the rates of URI on the two unit types were not statistically different. CONCLUSIONS: This study suggests that an inherent risk of ASCUs is an increased exposure to highly contagious infections such as upper respiratory infections. An intervention program effective in decreasing this risk to the level of traditional nursing units is proposed. A prospective study is needed to confirm these findings. PMID- 7490384 TI - High density lipoprotein cholesterol is associated with serum cortisol in older people. AB - OBJECTIVE: To determine the associations between serum cortisol and HDL cholesterol, other lipoprotein lipids and cardiovascular risk factors, carotid atherosclerosis, and clinical heart disease in older people. DESIGN: A cross sectional, observational, ancillary study of the Cardiovascular Health Study (CHS). POPULATION: A total of 245 community-dwelling people, 65 to 89 years old, were recruited consecutively for a 2-month period from the CHS cohort in Forsyth County, North Carolina. METHODS: Cortisol was measured by radioimmunoassay in serum collected between 7:00 and 10:00 AM after an overnight fast. Cortisol levels were correlated with lipoprotein lipids, insulin, glucose, body mass index, waist-hip ratio, prevalent coronary heart disease, hypertension, diabetes, and carotid atherosclerosis by B-mode ultrasound. RESULTS: Serum cortisol was correlated negatively (r = -.24) with body mass index and waist-hip ratio (r = .16) but was not related significantly to fasting insulin or glucose. Cortisol was not associated significantly with triglyceride and low density lipoprotein cholesterol but showed a positive correlation (r = .21) with high density lipoprotein cholesterol. The relationship between cortisol and high density lipoprotein cholesterol persisted after adjustment for gender, body mass index, waist-hip ratio, cigarette and alcohol use, triglyceride level, and diabetes. There was a trend toward a negative correlation between cortisol and measures of carotid atherosclerosis, but no significant relationship was indicated between cortisol and prevalent coronary heart disease, hypertension, or diabetes. CONCLUSION: Endogenous glucocorticoid levels correlated with HDL cholesterol levels and may play a role in the physiologic regulation of high density lipoprotein levels in older people. PMID- 7490385 TI - Insulin-like growth factor 1 and functional status in healthy older men. AB - OBJECTIVE: To determine if insulin-like growth factor 1 (IGF-1) is associated with strength and functional ability in healthy older men. DESIGN: Cross sectional study. SETTING: San Francisco Department of Veterans Affairs Medical Center. PARTICIPANTS: One hundred four ambulatory community-dwelling men. MEASUREMENTS: Serum IGF-1 levels were obtained. Measured variables included strength of the knee flexors and extensors, handgrip, score on the Physical Performance Test, body composition, and three tests of cognitive function. RESULTS: The subjects' mean age was 75.5 +/- 4.9 (SD) years (range 70-94 years), and their mean IGF-1 level was 134.7 +/- 43.6 ng/mL. The univariate association of age with the variables was much stronger than the univariate association of IGF-1 with the same variables. In multivariable models, age, but not IGF-1, was associated with the variables. CONCLUSION: In this study of healthy older men, age is the most important variable in predicting functional decline. There was no association of IGF-1 levels to functional status independent of age. PMID- 7490386 TI - Functional Incidental Training, mobility performance, and incontinence care with nursing home residents. AB - OBJECTIVE: To determine if an exercise intervention, Functional Incidental Training (FIT), results in improvements in mobility endurance and physical activity when compared with prompted voiding (PV) among cognitively and mobility impaired nursing home residents. DESIGN: Residents from four nursing homes were randomized into either a PV only (PV) or a PV plus FIT (FIT) intervention group for 8 weeks. Research staff implemented all intervention and measurement protocols. PARTICIPANTS: Seventy-six incontinent nursing home residents completed all phases of the trial. MEASURES: The standing, walking, and wheelchair endurance, physical activity, and frequency of agitation of all residents were assessed before, during, and after the 8-week intervention. RESULTS: The average length of time that subjects could walk or wheel was 2.6 and 4.6 minutes, respectively, at baseline. There was a significant group x time interaction after intervention, with only the FIT group showing improvements in walking, wheelchair, and standing endurance (Manova F = 4.56, 2.62, and 5.98, respectively; P < .05 in all cases). The frequency with which agitation was observed showed a significant drop over time in both groups (F = 14.3, P < .001), with no significant group x time interaction. CONCLUSION: The FIT intervention, which requires 6 minutes more nurses' aide time than does PV, increases both physical activity and mobility endurance in extremely frail and deconditioned nursing home residents. The increased cost of this intervention must be evaluated both in terms of clinical outcomes and by the reality that the target group for this intervention is very frail and will continue to require nursing home care, even assuming an excellent response to the intervention. PMID- 7490387 TI - Use of the Minimum Data Set to rate incontinence severity. AB - OBJECTIVES: To assess the relationship of the MDS incontinence severity ratings to direct measures of incontinence frequency. DESIGN: Two methods of measuring incontinence were compared: the MDS rating as recorded by nursing home (NH) staff and physical checks for wetness performed by research staff. SETTING AND PARTICIPANTS: A total sample of 293 older residents from nine nursing homes located in Iowa, the State of Washington, and the Los Angeles area were assessed once. A subsample of 49 incontinent residents were assessed twice, before and after the implementation of a prompted voiding program. INTERVENTION: This study is a part of a larger study evaluating the use of a computer-aided incontinence management system (IMS). Incontinent residents were assessed, and, if they met predefined criteria, they were treated with prompted voiding. MEASUREMENTS: Three measures of incontinence were used: the MDS rating recorded by NH staff, physical checks for wetness performed by NH staff while assessing residents for and treating them with prompted voiding, and physical checks for wetness performed independently by research staff. RESULTS: There was a statistically significant correlation (r = .49; P < or = .001) between research staff wet checks and the MDS ratings, but wetness checks performed by NH staff had an insignificant correlation with MDS ratings (r = .003; P < 0.914). There was wide variability within and between NHs in the correlation. For residents who were placed on the prompted voiding program, the pre to post wet rate, as measured by research staff, improved significantly (from 28 to 14%; t = 6.73; df = 48; P < .001), whereas the pre to post MDS ratings did not change significantly (from 1.7 to 2.0; t = -1.42; df = 48; P < .075). CONCLUSIONS: Although the MDS appears to identify incontinent NH residents accurately, its clinical utility may be limited by disagreements between actual wet check data and MDS categorical severity rankings for residents known to be incontinent. The wide variability between direct observational measures of wetness and the MDS scores denoting incontinence severity we observed may limit the potential usefulness of the MDS for detecting changes in incontinence severity. It is possible that more information and instructions are needed for staff completing the MDS if the goal is to discriminate between different levels of incontinence severity and measure changes over time in response to therapeutic interventions. PMID- 7490388 TI - Initial onset of accessory pathway-mediated and atrioventricular node reentrant tachycardia after age 65: clinical features, electrophysiologic characteristics, and possible facilitating factors. AB - OBJECTIVE: To evaluate the clinical features electrophysiologic characteristics, and possible facilitating factors in older patients (> or = 65 years) with initial onset of accessory pathway-mediated and atrioventricular (AV) node reentrant tachycardia. DESIGN: Of the patients undergoing electrophysiologic study and radiofrequency catheter ablation of accessory pathway-mediated and AV node reentrant tachycardia at this institution, patients with initial tachyarrhythmia after age 65 years were compared with those presenting initially before age 30. SETTING: A tertiary medical center for the general public. PARTICIPANTS: Sixty-six patients had their initial symptoms after age 65: Group I, 32 patients with accessory pathway-mediated tachycardia, and Group II, 34 patients with AV node reentrant tachycardia. Four-hundred forty patients had their initial symptoms before age 30: Group III, 283 with accessory pathway mediated tachyarrhythmia, and Group IV, 157 with AV node reentrant tachycardia. INTERVENTION: All patients underwent electrophysiological study to determine the mechanisms of tachyarrhythmia, and radiofrequency catheter ablation for treatment of tachycardia. RESULTS: (1) Older patients with initial arrhythmia had incidence of critical clinical manifestations, including tachyarrhythmia-related syncope and cardioversion, similar to those with initial arrhythmia at a younger age. (2) Patients in Group III, showed anterograde effective refractory period (ERP) of the AV node (P = .432), longer anterograde ERP of the accessory pathway (P = .004), and greater difference of the anterograde ERP between the AV node and the accessory pathway (D-ERP) (P = .003) similar to patients in group I. In Group II, the ERP and Wenckebach cycle length of the retrograde fast pathway was significantly longer than in Group IV (P = .037 and P < .001, respectively), and a greater percentage of patients in Group II than in Group IV AV node reentrant tachycardia needed isoproteronol to facilitate the induction of reentrant tachycardia (P = .034). (3) Patients in Group I and Group II had a higher incidence of supraventricular and ventricular ectopic activity than those in Group III (P = .002 and P = .005, respectively) and Group IV (P = .024 and P = .012, respectively) in 24-hour ambulatory electrocardiograms. CONCLUSION: The initial onset of accessory pathway-mediated tachycardia after age 65 may be caused by changes of electrophysiologic properties (greater D-ERP) as well as increased supraventricular and ventricular ectopic activity. Influence of the autonomic nervous system, rather than changes of conduction properties in the AV node, and increase in ectopic activity may contribute to the new onset of AV node reentrant tachycardia in older adults. The choice of antiarrhythmic drugs and radiofrequency ablation require attention to the clinical profile and facilitating factors of reentrant tachycardia in this group of patients. PMID- 7490389 TI - Health services use and mortality among older primary care patients with alcoholism. AB - OBJECTIVE: To describe the prevalence of alcoholism in an older primary care population and to compare rates of health services use and mortality among those with and those without evidence of alcoholism. DESIGN: Baseline screening for alcoholism using the CAGE questionnaire and longitudinal assessment of health services use and mortality using an electronic medical record system. SETTING: An academic primary care group practice at an urban ambulatory care clinic. PATIENTS: A total of 3954 patients aged 60 and older who completed the CAGE alcoholism screening questionnaire during routine office visits. MAIN OUTCOME MEASURES: Comorbidity, preventive health services use, hospital episodes and length of stay, emergency room visits, ambulatory care visits, total outpatient charges, and mortality. RESULTS: The prevalence of current evidence of alcoholism, as defined by a CAGE score > or = 2 and alcohol use in the previous 12 months, was 10.6%. Patients with evidence of alcoholism were younger (66.2 vs 68.3 years), had fewer years of education (8.4 vs 9.1), were more likely to be male (65.8 vs 27.2%), black (71.2 vs 62.6%), smokers (40.4 vs 26.3%), and malnourished (32.5 vs 26.3%). Patients with alcoholism were more likely to have a diagnosis of obstructive lung disease (22.9 vs 18.3%), injuries (14.2 vs 8.3%), and gout (6.7 vs 2.9%) and less likely to have a diagnosis of hypertension (56.9 vs 61.8%), arthritis (23.3 vs 29.3%), and diabetes (15.9 vs 23.3%). Among those with evidence of alcoholism, 41.6% had a diagnosis of alcoholism in their outpatient medical record. Rates of completion of preventive health services did not differ between the two groups, and there was no difference in the number of ambulatory care visits, emergency room visits, or total outpatient charges. Patients with evidence of alcoholism were more likely to be hospitalized (21.5 vs 16.9%) and more likely to die within 2 years (10.6% vs 6.3%). CONCLUSIONS: One of 10 older patients in this primary care practice had current evidence of alcoholism, fewer than half of whom had documentation of alcohol abuse in their medical records. These patients were more likely to be hospitalized and more likely to die but did not consume a greater amount of outpatient resources. Further research is needed to determine if interventions to reduce alcohol use would also reduce excess hospitalizations and mortality among these older patients. PMID- 7490390 TI - Dental, visual, auditory and olfactory complications in Paget's disease of bone. AB - OBJECTIVE: To determine the prevalence of dental problems in Paget's patients and in a control population without Paget's disease. The relationship of localization of bony involvement of Paget's disease with the prevalence of dental, auditory, visual, and smell changes is examined. DESIGN: Cross-sectional national survey. PARTICIPANTS: Four hundred ninety-eight subjects with Paget's disease were generated randomly from the mailing list of the Paget Foundation. INTERVENTION: Each subject was sent a cover letter, two questionnaires, and a stamped, addressed return envelope. One questionnaire contained questions specific for the individual with Paget's disease. Subjects were asked to give the other questionnaire, which contained the same questions except for those asking about the specifics of Paget's disease, to an unaffected spouse or a similar aged friend. MEASUREMENTS: All subjects were asked about their age, gender, race, educational level, income, living situation, chronic medical conditions, self rating of their general physical and dental health, the number of teeth present, and the presence of dental, auditory, visual, and olfactory problems. Paget's subjects were also asked to indicate which bones were affected. RESULTS: Paget's disease subjects assessed their oral health to be poorer than the controls. Paget's subjects were more likely to report pain when opening the mouth and were more likely to have had dental extractions. A significantly higher percentage of Paget's individuals with facial or maxillomandibular involvement reported having heart trouble. Paget's individuals with skull, facial or maxillo-mandibular involvement were more likely to report a change in their hearing, sight, and smell than Paget's subjects without involvement of the craniofacial complex. Dental problems were reported by 93% of those Paget's subjects with maxillomandibular involvement, compared with only 10% for those with skull involvement only or involvement at sites distant to the craniofacial complex. CONCLUSIONS: Patients with Paget's disease of bone involving facial or maxillo mandibular parts of the skeleton have a higher prevalence of change in hearing, sight, smell, and dental problems. We also have found an association of heart disease in patients who have involvement of facial or maxillo-mandibular bones. PMID- 7490391 TI - Nosocomial transmission of epidemic keratoconjunctivitis to food handlers in a nursing home. PMID- 7490392 TI - Weight change and physical function in older women: findings from the Nun Study. AB - OBJECTIVE: To investigate the association between change in weight and decline in physical function in older women. DESIGN: Longitudinal study of a defined population of Catholic sisters (nuns) whose weight and function were assessed twice, an average of 584 days apart. SETTING: Unique life communities (convents) located throughout the United States. PARTICIPANTS: 475 Catholic sisters who were 75 to 99 years of age (M = 82.1, SD = 4.8) and were independent in at least one Activity of Daily Living (ADL) at the first assessment of weight and function. INTERVENTIONS: None. MEASUREMENTS: At each assessment, weight, ADLs, and cognitive function were evaluated as part of the Nun Study--a longitudinal study of aging and Alzheimer's disease. Annual percent weight change was calculated using weights from the two assessments, as well as the number of days that elapsed between assessments. RESULTS: Mean weight at first assessment was 140 pounds (range 78 to 232, SD = 27). The mean annual percent weight change was 0.1% (range 22% loss to 16% gain, SD = 3.8). Age- and initial weight-adjusted findings indicated that those participants with an annual percent weight loss of 3% or greater had 2.7 to 3.9 times the risk of becoming dependent in each ADL, compared to the sisters with no weight change. The elevated risk persisted in those who were mentally intact or were independent in their eating habits. CONCLUSION: Monitoring of weight may be an easy and inexpensive method of identifying older individuals at increased risk of disability. PMID- 7490393 TI - Physician-reported determinants of screening mammography in older women: the impact of physician and practice characteristics. AB - OBJECTIVE: Despite having markedly higher breast cancer risk, compliance of older women with screening mammography has been poor. This study was undertaken to determine which physician and patient practice characteristics were associated with high self-reported mammography referral rates for older women. METHODS: Primary care physicians (n = 129) from three socioeconomically diverse communities in Los Angeles were surveyed. Agreement with annual screening and self-reported referral rates were assessed for two groups of women, those 65 to 74 years of age and those 75 years and older. Screening outcomes were compared with physician and patient practice characteristics using bivariate and multivariate techniques. RESULTS: Although 73% of physicians agreed with annual screening of women aged 65 to 74 years, only 24% of physicians reported actually screening most women seen in this age group. Similarly, 57% of physicians agreed that women age 75 years and older should be screened annually, but only 21% reported recommending mammograms for most women seen in this age group. In multivariate analysis, white physicians (adjusted OR = 9.1), younger physicians (adjusted OR = 3.85), and those who used the American Cancer Society's low cost mammography projects (adjusted OR = 5.01) were more likely to report screening the majority of women seen. DISCUSSION: This study suggests that although physicians' intentions to screen older women may be relatively high, a gap exists between intentions and what is reported to be accomplished in practice. Race/ethnicity and physician specialty were the two strongest predictors of high self-reported referral rates, suggesting that targeted interventions may be useful. PMID- 7490394 TI - Psychotropic use among older residents of board and care facilities. AB - OBJECTIVE: To describe patterns of psychotropic use as well as clinically relevant and concurrent nonpsychotropic use among older residents of board and care facilities in 10 states. METHODS: Weighted use rates were based on data from a sample of 2054 residents aged 65 and older from 410 facilities in 10 states, drawn via a complex, multistage sampling design. Data, including drug name, use frequency, and PRN use, were collected on all drugs taken by the resident during a 7-day period. During face-to-face interviews with residents and/or proxies, data were also collected on resident characteristics. SUBJECTS: The majority of subjects were female, white, and widowed. Average age was 82 years. Approximately 25% had a current mental or emotional condition, and 8% had been hospitalized for a psychiatric condition during the year before admission. More than half needed help with at least one activity of daily living. Forty percent had moderate or severe cognitive impairment, and 18% had received mental health care during the previous year. ANALYSES: Weighted descriptive analyses were performed with Software for Survey Data Analysis (SUDAAN), which accounts for the complex, multistage sampling design. RESULTS: Approximately 35% of the older residents used at least one psychoactive agent, with 30% of psychotropic users receiving two to four different psychotropic medications. Use rates decline with age, for psychotropics overall and for therapeutic classes. Results revealed potentially problematic polypharmacy in relation to drug duplication within therapeutic classes, use of multiple psychotropics across classes, and concurrent nonpsychotropic use. CONCLUSIONS: Compared with community-dwelling older adults, this study revealed high psychotropic use rates among older board and care facility residents. Overall, our results suggest that serious consideration be given to the apparent need for systematic drug utilization review, a potentially useful program that is mandated in other settings. PMID- 7490395 TI - Hyponatremia in a nursing home population. AB - OBJECTIVE: To determine the prevalence of hyponatremia in a nursing home population and to identify clinical factors that increase the risk for development of hyponatremia. DESIGN: Retrospective and prospective record review. SETTING: A Veterans Affairs nursing home care unit. PATIENTS: One hundred nineteen residents, who ranged in age from 60 to 103 years. Sixty ambulatory patients, 62 to 91 years of age, who attended a geriatric medicine outpatient clinic served as a reference population. MEASUREMENTS: Most recent serum sodium, creatinine, BUN, and all serum sodium determinations during the preceding 12 months; clinical diagnoses, diet, medications, and significant events at the time of recorded hyponatremic episodes; response to acute water loading in a subset of patients; number of deaths in the 12 months following entry into the study. RESULTS: In the 119 nursing home patients, ages 60 years or older, the most recent serum sodium identified 18% who were hyponatremic, compared with a prevalence of 8% in similarly aged ambulatory patients. When all serum sodium determinations for the previous 12 months were examined, 53% of the nursing home patients had at least one episode of hyponatremia during this time period. There was a high incidence of central nervous system (CNS) and spinal cord disease in the total nursing home population. Episodes of hyponatremia were frequently associated with an increased intake of fluids, given either orally or intravenously, and with tube feeding. Water load testing revealed abnormal water handling consistent with the syndrome of inappropriate antidiuretic hormone (ADH) secretion in 18 of 23 patients who had a history of hyponatremia. Seventeen percent of the patients with hyponatremia died over the following 12 months, compared to a death rate of 21% in the normonatremic patients. CONCLUSIONS: Hyponatremia is a common occurrence in nursing home residents and may be a consequence of abnormal ADH secretion resulting from CNS disease. Exposure to increased fluid intake, or to a low sodium tube-feeding diet, can lead to the onset of hyponatremia or to a worsening of an already present low-serum sodium concentration. PMID- 7490396 TI - The importance of oral health in the older patient. AB - Oral health is important to general health because stomatologic disease affects more than the mouth. Increasing preservation of teeth among present and future cohorts of older people has increased their risk for serious disease from oral pathogens. The intent of this paper is twofold: first, to alert non-dental health personnel to the significance of oral health and oral disease in the older adult; and second, to recruit the assistance of non-dental professionals in helping patients to achieve and maintain an optimal oral condition. Normative aging processes alone have little effect on the oral cavity, but common disease processes affecting oral health include tooth loss, dental caries, periodontal diseases, and oral mucosal diseases (including candidiasis and squamous cell carcinoma). Systemic diseases and their treatments frequently affect salivary, oral motor, and oral sensory functions. As a result of bacteremia or aspiration of oral contents, organisms of oral origin can be responsible for serious nonstomatological infections. Clinicians caring for older people need to recognize the importance of stomatological health, include an oral component in the multidisciplinary geriatric assessment, support the education of patients on aspects of dental health, and advocate the expansion of personal and public oral health benefits for older adults. PMID- 7490397 TI - Fairhill guidelines on ethics of the care of people with Alzheimer's disease: a clinical summary. Center for Biomedical Ethics, Case Western Reserve University and the Alzheimer's Association. AB - These guidelines summarize the content of meetings of family caregivers and individuals with dementia of the Alzheimer's type who identified and spoke on ethical issues in dementia care and who engaged in dialogue with an interdisciplinary and interprofessional group of individuals working in the field of Alzheimer's disease. This inductive method begins with attentive listening to the voices of the affected population and family members, in contrast with a theoretical and deductive approach to ethics. PMID- 7490399 TI - When the bill comes due for the autonomy of demented older adults, who pays? PMID- 7490398 TI - Attitudes of beginning medical students toward older persons: a five-campus study. The University of California Academic Geriatric Resource Program Student Survey Research Group. AB - OBJECTIVE: To examine the attitudes of beginning medical students toward older persons and their medical care. DESIGN: Cross-sectional survey. SETTING: Five University of California medical schools. PARTICIPANTS: Beginning medical school students from the Class of 1994. MEASUREMENTS: Demographic characteristics, personal contacts with older persons, and previous coursework or research experience with older persons were assessed. Knowledge with regard to aging was measured with a group of 10 questions, which were modified from the Facts on Aging Quiz. The attitudes of students toward older persons were assessed using the Aging Semantic Differential (ASD), the Maxwell-Sullivan Attitude Scale (MSAS), and two case scenarios. RESULTS: Ninety-two percent of the participants (554 of 603) responded; 93% of these responses were usable. Students were much less likely to admit an acutely ill 85-year-old woman to an intensive care unit, intubate her, and treat her aggressively than they were to treat an acutely ill 10-year-old girl with underlying chronic leukemia. In multivariate models, male gender and younger student age were both independently predictive of less favorable attitudes regarding a 70-year-old person on the ASD Instrumental Ineffective subscale. Asian-Americans and males had less favorable attitudes on the Autonomous-Dependent and the Personal Acceptability-Unacceptability subscales. More knowledge with regard to aging was predictive of more favorable attitudes on the Autonomous-Dependent and Personal Acceptability-Unacceptability subscales. Male gender and Asian-American ethnicity were significantly associated with less favorable attitudes on the two scales derived from the MSASs. Students who had previously visited a long-term care facility were more likely to disagree that it takes too much time to care for older persons. CONCLUSION: Beginning medical students have already formed some unfavorable attitudes about older persons. Few independent predictors (either sociodemographic or students' previous experiences) of student attitudes could be identified that would help in the selection of students who had more favorable attitudes toward older persons. Hence, attempts to generate physicians with good attitudes must rely on curricular efforts during medical school and residency training. PMID- 7490400 TI - Fitness and function at the end of life. PMID- 7490402 TI - Effectiveness of oral antibiotic treatment in nursing-home acquired pneumonia. PMID- 7490404 TI - Predictors of motor vehicle crashes in a dementia clinic population: the role of gender and arthritis. PMID- 7490405 TI - New onset heart failure in a geriatric patient. PMID- 7490403 TI - Risk factors for pneumonia in nursing home residents. PMID- 7490401 TI - Reading the tea leaves of psychotropic drug utilization evaluation. PMID- 7490406 TI - Risk factors for appointment failures in a geriatric outpatient clinic. PMID- 7490407 TI - New version of brief depression scale. PMID- 7490408 TI - Fatal hypocalcemia from sodium phosphate enemas. PMID- 7490409 TI - Severe hyponatremia associated with ramipril therapy in an old woman. PMID- 7490410 TI - Amiodarone and phenytoin: interaction. PMID- 7490411 TI - Wouldn't Charles Prentice be pleased? PMID- 7490412 TI - Challenges facing optometric education. PMID- 7490413 TI - Dry eye among males infected with the human immunodeficiency virus. AB - BACKGROUND: Keratoconjunctivitis sicca (KCS) has been reported as a potential ocular manifestation of human immunodeficiency virus (HIV) infection. This study was designed to estimate prevalence of dry eye among HIV-infected males and correlate subjective and objective findings with disease severity. METHODS: All subjects and controls completed the McMonnies Dry Eye Questionnaire (MDEQ) and underwent diagnostic testing for dry eye including biomicroscopic assessment of the anterior segment, lacrimation kinetics, sodium fluorescein break up time, and vital staining with sodium fluorescein and rose bengal. RESULTS: A 38.8 percent prevalence of dry eye was found in the study group. Certain risk factors may be more likely to produce KCS than others and may be a better indication of potential dry eye than disease severity. CONCLUSIONS: Patient symptoms are not adequate predictors of dry eye, indicating the need for KCS diagnostic testing as part of a comprehensive ocular examination for HIV-infected individuals. PMID- 7490415 TI - The dizzy patient: a review of etiology, differential diagnosis, and management. AB - BACKGROUND: Dizziness is a lay term used to describe a variety of sensations. Unfortunately, the term "dizziness" does not have a precise medical definition, and additional information is typically required to further define the patient's problem. METHODS: When dizziness is a presenting complaint, distinctions must be made between vertigo (a sense of false movement), near-syncope (a feeling of impending faint), disequilibrium (loss of balance), and ill-defined lightheadedness (an inability to concentrate or focus the mind). Possible causes of dizziness include conflicts between visual and vestibular information, vascular problems, adverse reactions to medication, psychological difficulties, systemic disease, and the effects of aging. RESULTS: Dizziness is a symptom of a physiological or psychological illness, and therefore management is typically directed toward treatment of the underlying illness. In some cases the cause of the dizziness cannot be found, however, or is untreatable. In these cases, management is directed toward symptom reduction. CONCLUSIONS: Dizziness is a relatively common problem that arises from a variety of causes. In many cases, optometrists can participate in the diagnosis and management of patients with complaints of dizziness. PMID- 7490414 TI - Clinical experience with piggyback contact lens systems on keratoconic eyes. AB - BACKGROUND: A piggyback contact lens system (PBCLS) is used when traditional lenses do not provide optimal vision or tolerance. This study assessed the use of PBCLS in keratoconic patients. METHODS: The charts of 205 keratoconic patients were retrospectively reviewed and 16 patients wearing PBCLS were identified. Visual acuity, average wearing time, and ocular complication data with PBCLS were analyzed. RESULTS: Vision and average lens wearing time were stable, if not improved, for the 16 patients when the data was compared to the patients' wear of rigid gas permeable contact lenses. Although two patients developed neovascularization and one patient developed giant papillary conjunctivitis, all pre-existing corneal complications resolved with PBCLS wear. Average Dk/L through the center and mid-periphery of the PBCLS were 8.4 +/- 1.2 x 10(-9) (mean +/- standard deviation) and 4.5 +/- 1.8 x 10(-9) cm2ml O2/sec ml mmHg (mean +/- standard deviation) respectively. No gross corneal edema was noted upon slit lamp observation. CONCLUSIONS: PBCLS can be beneficial in the management of keratoconus. PMID- 7490416 TI - Chiasmal syndrome. AB - BACKGROUND: Chiasmal syndrome is the constellation of signs and symptoms that are associated with lesions of the optic chiasm. Pituitary adenoma is its single most common cause. METHODS: A MEDLINE search of the biomedical literature was conducted to uncover papers on topics related to chiasmal syndrome published since 1989. RESULTS: The MEDLINE search retrieved a total of 163 citations. Each citation was screened for relevance. Copies of each relevant paper were obtained and their references were examined for any publications that the original search may have missed and for older publications that continue to be of importance. CONCLUSIONS: This review article discusses the etiology, diagnosis, and management of chiasmal syndrome. PMID- 7490418 TI - Presidential inaugural address 1995--conquering challenges, creating successes. PMID- 7490417 TI - Atypical herpetic endotheliitis. AB - BACKGROUND: Three patients ranging in age from 26 to 40 years presented with unilateral posterior stromal lesions with diffuse stromal edema. There was no history of prior surgery, trauma, infectious disease, dystrophy or exposure to noxious agents. Only one patient had signs of anterior chamber reaction and none of the patients showed any sign of epithelial involvement or of associated blood vessels or scarring. All three patients presented with features atypical of herpetic disease, such as lack of epithelial involvement and posterior stromal opacification. They also demonstrated many of the typical characteristics of herpetic disease, however, and thus were diagnosed as having herpetic endotheliitis. RESULTS: All three patients were treated with a regimen of antiviral agents and corticosteroids. Two patients improved, although the time frame differed in each. One patient subjectively improved, then worsened 1 month later after discontinuing treatment on her own. She was lost to subsequent follow up. CONCLUSIONS: Young patients presenting with unilateral posterior stromal opacification and stromal edema in the absence of epithelial involvement are likely to have endotheliitis of herpetic origin. PMID- 7490419 TI - Co-localization of NADPH diaphorase reactivity and vasoactive intestinal polypeptide in human colon. AB - Nitric oxide-containing nervous structures were localized in the human colon using NADPH diaphorase activity and nitric oxide synthase immunoreactivity. We found some, solitary NADPH diaphorase-reactive and nitric oxide synthase immunoreactive neurons in the submucous plexus, while the myenteric plexus contained several neurons, often arranged in clusters, and nerve fibers showing these markers. The circular muscle layer contained a dense plexus of NADPH diaphorase-reactive nerves, which was greater than that in the longitudinal muscle layer. We report on co-localization of NADPH diaphorase activity and VIP immunoreactivity in several neurons of the myenteric ganglia. Such co localization has not been reported previously for human colon. Localization of nitric oxide synthase and VIP in the myenteric plexus and in the nerves of circular muscle layer raises the possibility that nitric oxide contributes to the regulation of motility in the human colon. PMID- 7490420 TI - Nerves containing nitric oxide synthase and their possible function in the control of catecholamine secretion in the bovine adrenal medulla. AB - NADPH-diaphorase reactivity and neuronal nitric oxide synthase (nNOS) immunostaining have been localised in sections of bovine adrenal glands. Both were present in nerve fibres and terminals in the subcapsular region and running between zona glomerulosa cells, amongst the medullary chromaffin cells, between large ganglion cells in rare encapsulated medullary ganglia and in large nerve bundles running through the cortex. Occasional isolated fibres were stained in deeper cortical layers. Both NADPH-diaphorase reactivity and nNOS immunoreactivity were present in a population of ganglion cells located individually or in small groups at the medullary-cortical boundary. NADPH diaphorase reactivity was also found in all cortical cells (zona glomerulosa cells being more densely stained than other cortical cells) and in large fibrous structures in large nerve bundles (tentatively identified as glial cells): these structures were not stained with antisera to nNOS. Chromaffin cells were not stained with either technique. The possible role of neurally-released nitric oxide in the regulation of nerve-induced catecholamine secretion from chromaffin cells was investigated in isolated, perfused, bovine adrenal glands. The secretion of both adrenaline and noradrenaline in response to field stimulation of adrenal nerves at either 2 Hz or 10 Hz was unaffected by the presence of N omega-nitro-L-arginine (30 microM), sodium nitroprusside (10 microM) or L arginine (100 microM) in the perfusing solution. It is concluded that, although nitric oxide may be generated and released from adrenal medullary nerves innervating chromaffin cells, it does not play a direct role in the acute regulation of adrenal catecholamine secretion. PMID- 7490421 TI - Choline acetyltransferase-immunoreactive neurones in a prevertebral sympathetic ganglion, the inferior mesenteric ganglion. AB - Using immunohistochemical techniques a small population of choline acetyltransferase (ChAT) immunoreactive (IR) neurones has been identified in the inferior mesenteric ganglion (IMG) of guinea pig (4.6% of all neurones), ferret (6.4%) and rat (0.4%). A detailed study in the guinea-pig IMG revealed that the vast majority of cholinergic neurones did not express tyrosine hydroxylase (TH) IR, indicating that they were non-catecholaminergic. The cholinergic neurones were significantly larger than the TH-positive neurones. The majority of the ChAT IR cells (64%) was observed in small clusters which were consistently located in the caudal lobe of the IMG close to the entry of the hypogastric nerves. 83% of the ChAT-IR cells also contained neuropeptide Y (NPY). Since the vast majority of TH-negative cells were ChAT-positive (94%), the TH negativity was taken as an indirect indication for ChAT-IR. NPY-IR, somatostatin (SOM)-IR and vasoactive intestinal peptide (VIP)-IR were found in both the TH-IR cells (22, 84 and 1%, respectively) and the putative cholinergic population (95, 84 and 70, respectively). Thus the majority of cholinergic neurones in the IMG were likely to contain NPY, SOM and VIP. TH-IR cells exhibited an extensive innervation of fibers immunoreactive for ChAT, VIP, ENK and NOS. In contrast, only a sparse plexus of ChAT-, ENK-, NOS-, NPY- and SOM-positive fibres was found around the TH negative cells. VIP-IR fibres did not appear to innervate ChAT neurones. PMID- 7490422 TI - Effect of oral ethanol intake on muscle sympathetic nerve activity and cardiovascular functions in humans. AB - The aim of the present study was to clarify the acute effect of alcohol drinking on muscle sympathetic nerve activity and other cardiovascular variables in young healthy human volunteers. Seven volunteers (25.0 +/- 4.7 years in age, weighing 59.9 +/- 5.9 kg) were instructed to lay down on a bed, and muscle sympathetic nerve activity (MSNA) was microneurographically recorded from the tibial nerve, simultaneously with an electrocardiogram, blood pressure with Finapres and autosphygmomanometer, cardiac output by impedance cardiography, and skin blood flow by laser Doppler flowmetry. After a 1-h rest, the subjects drank alcohol (0.6 g/kg) and were remained in resting position for 105 min. Blood ethanol levels indicated that they were moderately intoxicated. Heart rate constantly increased until 30 min after the ingestion, and maintained a peak level. MSNA was slightly but significantly suppressed just after the ingestion, and was gradually enhanced until the end of the experiment, showing a significant difference from the control level until 40 min in burst rate and until 25 min in total MSNA after the ingestion. Blood pressure showed an abrupt and transient increase at first, then gradual decrease until 30 min after the ingestion, and maintenance of the resting level until the end of the experiment. Cardiac output showed no constant tendency and no significant differences after the ingestion with wide interindividual variation. Skin blood flow increased 15 min after the ingestion.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7490423 TI - Influence of spinal cord injury on the morphology of bladder afferent and efferent neurons. AB - Severe micturition dysfunction can occur following spinal cord injury (SCI) due to abnormal contractions of the urethral sphincter during bladder contractions (bladder/sphincter dyssynergia). This causes urinary retention, bladder overdistension, and increases the workload of the bladder leading to hypertrophy of the bladder muscle. Bladder hypertrophy induced by urethral outlet ligation in rats is accompanied by enlargement of both the afferent and efferent neurons innervating the bladder. The primary aim of this study was to test whether SCI induced bladder hypertrophy produces a similar enlargement of bladder afferent neurons in the dorsal root ganglia (DRG) or efferent neurons in the major pelvic ganglia (MPG). Following SCI in female Wistar rats, there was a four-fold increase in bladder weight. The mean cross-sectional area of bladder DRG cell profiles increased approx. 50% after SCI; however, the mean area of MPG cell profiles did not change significantly. Urinary diversion (disconnecting the ureters from the bladder) prevented both the bladder hypertrophy and the DRG cell hypertrophy after SCI, suggesting that bladder hypertrophy drives DRG cell enlargement. On the other hand, since the size of MPG cells did not change significantly after SCI, bladder hypertrophy does not mandate MPG cell enlargement. However, preliminary results indicate that the mean cross-sectional area of MPG cells did increase (2-3 times) in SCI rats when the neural input to the MPG was eliminated by transecting the pelvic and hypogastric nerves; this suggests that the lack of change in size of MPG cells after SCI may be due to an inhibitory influence from the spinal cord. PMID- 7490424 TI - Assessment of autonomic function in traumatic quadriplegic and paraplegic patients by spectral analysis of heart rate variability. AB - We analyzed by means of autoregressive spectral analysis the spontaneous beat-to beat heart rate variability (HRV) of quadriplegic and paraplegic male subjects at rest in the supine position. In agreement with our previous study, in nine of 15 quadriplegic patients only the high-frequency (HF: center frequency = respiratory frequency) component (a marker of vagal modulation of heart rate) was observed. In contrast, in six of the quadriplegic patients both the HF component and the low-frequency (LF: center frequency at approx. 0.1 Hz, 0.03-0.15 Hz in this study) component (a marker of sympathetic and vagal modulation of heart rate) were observed. However, in six quadriplegic patients who presented the LF component, (i) the center frequency of the LF component was lower than that in 10 healthy, sedentary, age-matched males (control I) (P < 0.01), (ii) the power of the HF component was smaller than that in the control-I group (P < 0.01) and (iii) the LF/HF power ratio (an index of sympathovagal balance) was larger than that in the control-I group (P < 0.05). On the other hand, in nine paraplegic patients with an intact 1st-4th thoracic spinal cord, from which the cardiac sympathetic nerves originate, the total power, the power of the LF component and that of the HF component were smaller than those in nine healthy, sedentary, age matched males (control II) (P < 0.05, P < 0.01 and P < 0.01, respectively).(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7490425 TI - Circadian and short-term variabilities in blood pressure and heart rate measured by telemetry in rabbits and rats. AB - Blood pressure (BP) and heart rate (HR) were measured by telemetry in conscious unrestrained rabbits to clarify the profile of their variabilities. The variabilities were assessed for two periods, 24 h (circadian rhythm) and 1 h (short-term variability), and compared with those in rats. BP and HR in rabbits were lower than those in rats but the circadian rhythms in rabbits showed nocturnal patterns as is the case in rats. In contrast, short-term variabilities in BP in rabbits were considerably larger than those in rats. The short-term variability in BP in rabbits was suppressed by alpha-adrenergic blocking without changes in basal values but not by beta-adrenergic blocking or angiotensin converting enzyme inhibition. These results demonstrate the need to take the unique characteristics of short-term variability in BP in rabbit into consideration when the circadian rhythm is focused on and that the short-term variability in BP in rabbits is caused mainly by activation of alpha-adrenergic receptors. PMID- 7490426 TI - Human pupillary light reflex during and after two-fold Valsalva maneuver. AB - The influence of the Valsalva maneuver on the pupillary light reflex and on the parameters of human pupillogram were studied in 9 normal volunteers (mean age 21.0 years). The consensual pupillary reactions to light were recorded using a infrared-reflecting pupillograph; pupillogram records were processed off-line by a computer system. Latency time increased during the Valsalva maneuver and normalized in the first minute after its end. No significant changes were found in constriction time. Half-redilatation time showed the opposite dynamics of latency time. The amplitude diminished during Valsalva maneuver and in the first minute after it. These results suggest a possible alteration of the autonomic nervous regulation during Valsalva maneuver and especially some changes in parasympathetic outflow to human iris sphincter. The possible effect of prevailing parasympathetic or sympathetic activity on pupillometry parameters during some phases of the Valsalva maneuver and after its end is discussed. PMID- 7490427 TI - Multifunctional ventral respiratory group: bulbospinal expiratory neurons play a role in pudendal discharge during vomiting. AB - Pudendal motoneurons are activated in phasic bursts during the retching and expulsion phases of vomiting. The resulting contraction of the anal and urethral sphincters serves to maintain continence during the large increase in abdominal pressure that occurs during vomiting. We evaluated the contribution of bulbospinal expiratory neurons located in the portion of the ventral respiratory group (VRG) caudal to the obex (nucleus retroambigualis) to the control of pudendal motoneurons during fictive vomiting in decerebrate, paralyzed cats. Pudendal nerve discharge is abolished by cutting the axons of caudal VRG expiratory neurons as they cross the midline between the obex and C1 before descending in the spinal cord. All caudal VRG expiratory neurons that were antidromically activated from the sacral spinal cord, where the pudendal motor pool (nucleus of Onuf) is located, discharged strongly during the end of the expulsion phase of vomiting. However, only a small proportion of these neurons was active in phase with pudendal discharge during the retching phase. The apparent involvement of caudal VRG expiratory neurons in the control of pudendal motoneurons during vomiting is another example of the multifunctional role that can be played by respiratory-related neurons in the mammalian nervous system. PMID- 7490428 TI - In vitro testing of antimicrobial agents. AB - Indications for determining the inhibitory activity of an antimicrobial agent are well defined, and the data provided by such testing are useful for guiding therapy in a cost-effective manner. Susceptibility test methods with inhibition as an endpoint are standardized, and guidelines regarding methodology, quality control, and reporting are published. The ultimate decision concerning reporting, however, should be made jointly by clinical microbiologists, members of the hospital Pharmacy and Therapeutics or Infection Control Committee, and infectious disease clinicians. The NCCLS also has published tentative guidelines regarding methods for determining bactericidal activity of antimicrobial agents. Given the many biologic and technical variables that influence these tests, standardization is critical for appropriate interpretation of results. In contrast to tests that measure inhibition, however, the clinical value of bactericidal testing is poorly defined, and additional information gained by conducting well-designed, prospective, blinded clinical trials is needed. Methods for determining synergism/antagonism have not been standardized, which makes interpreting results and comparing results of tests performed at different institutions difficult. With the exception of detecting high-level aminoglycoside resistance in enterococci as a means to predict synergism between these drugs and cell wall active agents, the clinical relevance of assessing synergism is not well documented. PMID- 7490429 TI - Pharmacodynamics of antimicrobial agents. Bactericidal and postantibiotic effects. AB - Pharmacodynamics of an antimicrobial agent relate the time course of concentrations of the drug to the antimicrobial effects at the site of infection. Antimicrobial agents can be divided into three main groups, based on antimicrobial pharmacodynamic characteristics: 1) agents that exhibit concentration-dependent bactericidal activity over a wide range of drug concentrations (e.g., the aminoglycosides and fluoroquinolones); 2) agents that exhibit time-dependent bactericidal activity that has little relationship to the magnitude of the drug concentrations, as long as the concentrations are above a minimally effective level (e.g., vancomycin and beta- lactam antibiotics); and 3) agents that exhibit predominantly a bacteriostatic effect. Knowledge of the antimicrobial pharmacodynamic characteristics provides a more rational basis for determination of optimal dosing regimens in terms of size of the dose and length of the dosing interval. PMID- 7490430 TI - Pathogens resistant to antimicrobial agents. Epidemiology, molecular mechanisms, and clinical management. AB - The emergence of resistance to antimicrobial agents continues to be a major problem among both nosocomial and community-acquired pathogens. Bacteria employ a variety of strategies to avoid the inhibitory effects of antibiotic agents, and have evolved highly efficient means for the dissemination of resistance traits. The result has been the emergence of multidrug-resistant pathogens such as penicillin-resistant pneumococci, vancomycin-resistant enterococci, methicillin resistant staphylococci, as well as a variety of multiresistant gram-negative organisms. Control of antibiotic-resistant pathogens will provide a major challenge for both the medical community and society in general. The implication of a failure to meet this challenge is the eventual arrival of the "post antibiotic era." PMID- 7490431 TI - Principles of selection and use of antibacterial agents. AB - Treatment of infection involves a complex interaction among the infecting organism (susceptibility to the therapeutic agent), host factors (immune function, site of infection, renal and hepatic metabolism), and pharmacokinetics (rate of absorption, distribution and excretion). Successful therapy requires careful consideration of these factors, and unsuccessful treatment should invoke a careful reanalysis of them. PMID- 7490432 TI - Topical antibacterial agents. AB - Topical antibacterial agents offer a useful alternative to systemic agents in certain circumstances. Uses include prophylaxis of infection for burns, traumatic wounds, and intravascular catheters, as well as eradication of S. aureus nasal carriage and treatment of primary and secondary pyodermas. Evidence supporting the use of topical agents for prophylaxis and treatment of skin and superficial wound infections and the indications for use of specific antimicrobials have been reviewed. Although topical agents are widely used, in many instances, data supporting their efficacy are only beginning to emerge. PMID- 7490433 TI - Antibacterial agents in pediatrics. AB - The utility of various antibacterial agents for therapy of infectious diseases in children is determined by the unique pharmacokinetics and potential toxicity in children. The important age-related principle of pharmacokinetics is reviewed in the first section of this article; the second section focuses on specific therapeutic agents and their use in children. Particular emphasis is given to the use of new antibiotics in children, including the new oral cephalosporins and macrolides. PMID- 7490434 TI - Antibiotic concerns in the elderly. A clinician's perspective. AB - Unique antibiotic concerns exist for the management of infections in elderly patients. Issues concerning age-associated physiological alterations, compliance, drug-drug interactions, adverse drug reactions, and guidelines for empiric therapy are reviewed in this article. PMID- 7490435 TI - Antibacterial agents in renal failure. AB - This article provides background information on the pharmacokinetics of antimicrobial agents in patients with normal and impaired renal function. Tables are provided to allow quick determination of appropriate dosages for varying degrees of renal failure. Specific mention is made of aminoglycoside dosing, dosage adjustment in dialysis, and antibiotic toxicity in renal failure. PMID- 7490436 TI - Antibacterial agents in infections of the central nervous system and eye. AB - Experimental animal models have provided much information that can be applied to antimicrobial therapy of infections of the central nervous system and eye. The efficacy of an antimicrobial agent in the therapy of meningitis depends upon its ability to penetrate the blood-brain barrier, be active in purulent cerebrospinal fluid, and demonstrate rapid bacterial activity against the offending pathogen. In ocular infections, topically administered drugs must overcome various barriers to penetrate into the eye, or these barriers must be bypassed (i.e., by periocular or intravitreal injection) for optimal therapy. This article reviews the basic therapeutic principles for the treatment of infections of the central nervous system and eye, and gives recommendations for the treatment of specific infections. PMID- 7490437 TI - Antibacterial agents in pregnancy. AB - Except for topical, nonabsorbable agents, any antibiotic chosen for use in pregnancy exposes the fetus as well as the mother to its effects. Therefore, initiation of antimicrobial therapy must be based on clear-cut necessity. Because the physiology of the maternal-fetal unit is complex and ethical consideration of potential fetal harm is pre-eminent, data on safety are derived from animal studies, incidental observations on individual women treated with an agent, or longitudinal tracing of groups of women who required treatment with a particular antimicrobial regimen. Consequently, a rating of absolute safety in pregnancy has never been assigned to any currently available antimicrobial agent. Decades of clinical experience with penicillins, cephalosporins, and erythromycins have documented the pharmacokinetics of these drugs in pregnant women as well as their overall safety for the fetus. These classes of drugs are those most favored for use in pregnancy for susceptible infections. Although aminoglycosides have known toxic effects on the fetus, they are safe to use if serum levels are carefully monitored in the mother. Agents in the quinolone, sulfonamide, and tetracycline categories should be avoided unless maternal necessity for their use justifies the exposure of the fetus to their toxicity. Both clinical and experimental data are very limited on the newer agents, such as the new macrolides, azithromycin and clarithromycin. The first-line agents for the treatment of TB (i.e., INH, rifampin, and ethambutol) are considered safe in pregnancy, but in the era of multidrug-resistant mycobacterial isolates, agents with known or suspected fetal toxicity may need to be used. PMID- 7490438 TI - Newer penicillins and beta-lactamase inhibitors. AB - Penicillins continue to be essential antibiotics for the treatment and prophylaxis of many infectious diseases. Recent advances have resulted in compounds with favorable new antimicrobial and pharmacologic properties. This article reviews the spectrum of activity, toxicity, pharmacokinetics, and clinical uses of the extended spectrum penicillins and the beta-lactamase inhibitor combination agents. PMID- 7490439 TI - The newer cephalosporins. Aztreonam and imipenem. AB - Many of the antimicrobial agents described here exhibit great advances over older drugs in terms of antimicrobial spectrum, clinical utility, and, sometimes, safety. The newer cephalosporins are useful for treatment of many common outpatient and inpatient infections. Aztreonam provides excellent coverage against a broad range of aerobic gram-negative bacteria, without the toxicity associated with aminoglycosides. Imipenem exhibits activity against an impressive array of pathogens. These antimicrobials are expensive, however, and some offer no advantages over older agents. Finally, all--including imipenem--are faced with increasing resistance of bacteria. PMID- 7490440 TI - Fluoroquinolones. AB - Since the introduction of the floroquinolones for clinical use in the late 1980s, they have been used successfully for a large number of clinical situations. As experience accumulates, the indications and optimal use of these agents gradually become more clear. Unfortunately, two of the pathogens for which these agents were most promising--methicillin-resistant S. aureus and P. aeruginosa--have developed resistance. Currently, the quinolones are excellent agents for the treatment of complicated urinary tract infections, including those caused by P. aeruginosa. In addition, they should be considered as initial therapy for the treatment of severe bacterial gastroenteritis. The quinolones should also be considered when attempting to eradicate the chronic stool carriage of S. typhi. These agents also offer significant advantages in the treatment of osteomyelitis and prostatitis caused by gram-negative bacilli that frequently require prolonged antimicrobial therapy. Treatment of STDs, especially gonorrhea, is another clear indication for their use. Ciprofloxacin should be considered as initial therapy in patients with malignant otitis externa and in cystic fibrosis patients with exacerbations secondary to P. aeruginosa in the sputum. The role of the quinolones for soft tissue and respiratory tract infections is less clear and their use probably should be limited to certain situations in which there is a clear advantage over beta-lactams, macrolides, and trimethoprim-sulfamethoxazole. The new quinolones, fleroxacin, perfloxacin, sparfloxacin, and tosufloxacin, which are being developed and tested for clinical use, will offer advantages in once-a-day dosing and better gram-positive antimicrobial activity. Because the inappropriate or heavy use of the fluoroquinoles has resulted in considerable development of resistance, it is imperative that they be used only when there is a distinct advantage over conventional therapy in terms of efficacy, safety, or cost. Otherwise, the rapid development of resistance will jeopardize the potentially bright future for this entire class of compounds. PMID- 7490441 TI - The newer macrolides. Azithromycin and clarithromycin. AB - Azithromycin and clarithromycin are structural analogues of erythromycin that have similar mechanisms of action. The newer macrolides have several distinct advantages over erythromycin, including improved oral bioavailability; longer half-life, allowing once or twice daily administration; higher tissue concentrations; and fewer gastrointestinal adverse effects. Clarithromycin and azithromycin also have enhanced antimicrobial activity. The clinical efficacy of the newer macrolides has been similar to erythromycin for the treatment of upper and lower respiratory tract and skin and soft tissue infections. New therapeutic roles include the use of azithromycin for C. trachomatis infections and the inclusion of clarithromycin or azithromycin as part of therapeutic regimens for disseminated MAC infections in HIV-infected patients. Further clinical trials are needed to determine the optimal roles for and uses of these new macrolides. PMID- 7490442 TI - Vancomycin, trimethoprim-sulfamethoxazole, and rifampin. AB - Vancomycin is a safe, effective antibiotic for a variety of serious gram-positive infections. Because of emerging vancomycin-resistant enterococci and the threat of spread of vancomycin-resistant genes to other gram-positive organisms, judicious use of vancomycin should be promoted. Trimethoprim-sulfamethoxazole, a broad-spectrum synergistic combination, continues to find widespread use in gram negative bacterial infections, especially involving the urinary tract. It has enjoyed enormous success in prophylaxis and therapy of Pneumocystis carinii pneumonia. Rifampin remains of paramount significance as a cidal agent against Mycobacterium tuberculosis; however, increasing reports of strains of MDR tuberculosis are of great concern. PMID- 7490444 TI - Antimicrobial prophylaxis of infection. AB - Antimicrobial agents are used to prevent infections in a variety of clinical circumstances. In certain instances, the precise indications for prophylaxis remain controversial, and the preferred regimens undergo alterations based upon evolving clinical experience, changing patterns of microbial susceptibility, and innovations in medical and surgical practice. This article outlines the general principles underlying the use of antimicrobial prophylaxis and presents recommendations for the use of such prophylaxis in three areas: (1) surgery involving contaminated, clean-contaminated, and clean procedures; (2) prevention of infections due to specific pathogens, including Neisseria meningitidis, Hemophilus influenzae, Streptococcus pneumoniae, and Streptococcus pyogenes; and (3) prevention of infective endocarditis. PMID- 7490443 TI - Adjuncts to antibacterial therapy. AB - Strategies of augmenting or attenuating the immune system's response to infection are promising potential methods to enhance antibacterial therapy. Many of these new therapies are natural cytokines, such as the hematopoietic growth factors and interferons, which upregulate the immune response. Other examples include replacement therapies, such as immune globulin, and downregulators of the immune response, such as corticosteroids. In the near future it may be possible to adjust the host response to microbial infection to maximally inhibit the microbe while minimizing inflammatory damage. PMID- 7490446 TI - The application of in vitro isolation, cryopreservation and patch-clamp microelectrode recording methods to adult rat thymic nurse cells. AB - Characterisation of the ionic mechanisms possessed by immune cells has begun to reveal a range of transmembrane ion channel properties which may have immunological significance. Since thymic epithelial cells appear to influence selection of the T cell repertoire, understanding their membrane physiology may be of importance. A method is therefore outlined for the targeting of patch-clamp electrophysiological measurements to acutely isolated, fresh and cryopreserved adult rat thymic nurse cells. These cells represent a discrete and predominantly cortical population of epithelium. Cells were separated by enzymatic and mechanical dispersal of thymus, enriched by sedimentation and identified on the basis of their characteristic lympho-epithelial, multi-cellular morphologies. Phase-bright cells retaining this anatomical form survived freezing and the voltage-gated conductances in such cells are described. The merits of this approach include the preparation and storage of mature differentiated phenotypes of immune cells for in vitro studies. Their preservation permits temporal separation of experiments, correlative experimentation on identical samples, optimises cell preparation in terms of cost and labour, and reduces animal and tissue requirements. PMID- 7490445 TI - A rapid non-selective method to generate quadromas by microelectrofusion. AB - A simple non-selective methodology was developed and standardized to generate desired hybrid-hybridoma or quadroma secreting bifunctional antibodies. This novel protocol is based on microelectrofusion on a meander chamber using a few hundred cells of each of the two parental hybridomas with no laborious drug selection procedures. Seeding approximately 10 cells per well in a 96-well microtitre plate after fusion in 200 microliters standard medium containing 20% FBS and 10% Origen growth factor generated positive quadromas secreting bispecific antibodies with good stability after the second reclone. Compared to the conventional PEG fusion and other methods this simple protocol is both rapid and economical. Generally, conventional methods to make quadromas and triomas require the introduction of drug selection markers into one or both of the parental cells, a procedure that could take 3-6 months. Utilizing the non selective microelectrofusion method described here, we have generated several quadromas in a very short time. Further, such a protocol could also be potentially adopted to generate human hybridomas with few B cells isolated from peripheral blood lymphocytes enriched by antigen specific panning or affinity microelectrofusions. PMID- 7490447 TI - Preparation and characterization of fluorescein-labeled plasma membrane vesicles from transfected Chinese hamster ovary cells. AB - Plasma membrane vesicles have been used to study a wide variety of biological events including the nature of receptor-ligand interactions and the physiology of molecular transport across membranes. Plasma membrane vesicles were chemically induced from an adherent Chinese hamster ovary cell line expressing recombinant glycophorin A, a well-studied intrinsic membrane protein of the red blood cell. They were also prepared from Chinese hamster ovary cells in suspension culture. Biochemical and immunological assays demonstrated the equivalence of glycophorin A on cell membranes and vesicles. The transfected Chinese hamster ovary cell membrane was also labeled with a highly aliphatic fluorescent cell linker. Vesicles produced by the fluorescein-labeled cells demonstrated bright surface staining of the plasma membrane. They too expressed glycophorin A biochemically and immunologically indistinguishable from cellular-based glycophorin A. Plasma membrane vesicles are non-adherent in culture and stably retain the fluorescent probe in the cell membrane. Fluorescein-labeled vesicles will be particularly useful for studying cellular interactions in which both constituents of a receptor-ligand pair are expressed on adherent cell lines. Unlabeled vesicles may also prove to be useful as soluble immunoadsorbants in both the clinical laboratory and basic science research settings. PMID- 7490448 TI - Quartz crystal microbalance detection of Vibrio cholerae O139 serotype. AB - A piezoelectric (PZ) quartz crystal microbalance (QCM) biosensor for the rapid detection of Vibrio cholerae serotype O139 has been developed. The antibody to this serotype was immobilized on the gold transducer surface of a 10 MHz AT cut PZ crystal. Solutions containing known antigen concentrations were then incubated for 1 h on the antibody-bound transducer. The biosensor was able to detect 10(5) cells per ml of O139 versus a background of O1 (Ogawa) serotype. PMID- 7490449 TI - Establishment and evaluation of MRK16-magnetic cell sorting assays for detecting low expression of multidrug resistance P-glycoprotein using human leukemia cell lines and peripheral blood cells from healthy donors. AB - Two types of magnetic cell sorting assays, termed MRK16-MACS and MRK16-MACS-FACS, have been established to detect low expression level of P-glycoprotein (P-gp) using a monoclonal antibody MRK16, which recognizes a cell surface epitope of P gp. With K-562 and U-937 cell lines, which are known to express low levels of P gp and hence routinely used as negative control cell lines in conventional flow cytometry, both assays gave significantly positive reactivities indicating improved specificity and sensitivity of these assays. The findings in the dilution test, where P-gp-positive cells were added to P-gp-negative cells at various ratios, demonstrated that the MRK16-MACS assay is quantitative and capable of detecting small numbers of P-gp-positive cells as few as 2.5% of the total cells tested. Furthermore, specific enrichment of P-gp-expressing cells in magnetic cell sorting assays was verified by reverse transcription-polymerase chain reaction (RT-PCR) analysis and functional assay for P-gp with Rhodamine 123. The availability of such magnetic cell sorting assays may offer an approach to quantitate low level of P-gp expression. PMID- 7490450 TI - Intrafamily fragment analysis of the T cell receptor beta chain CDR3 region. AB - The length distributions of the third complementarity determining region (CDR3) of the T cell receptor (TCR) beta chain were determined quantitatively from peripheral blood lymphocytes of healthy donors. RT-PCR products of 26 V beta families and subfamilies were analyzed by an A.L.F. DNA sequencer and Fragment Manager software. We established a normal reference of CDR3 lengths for most of the V beta families and subfamilies. The known range of CDR3 lengths for the V beta chain was expanded to 24 amino acids, and quantitative measurements were made for each fragment length allowing intrafamily CDR3 fragment length comparisons. Importantly, we were able to analyze intrafamily CDR3 fragment distribution without optimizing PCR conditions, thereby circumventing a major obstacle found in intrafamily TCR beta chain comparisons. PMID- 7490451 TI - A method for preparing chelate-cytokine conjugates with retention of protein structure, biological activity, and pharmacokinetic properties. AB - The chelating agent diethylenetriaminepentaacetic acid (DTPA) has been conjugated site-specifically to the N-terminus of recombinant human interleukin-2 (rhIL-2) by reaction with DTPA dianhydride at an initial pH of 6.0, thus demonstrating broader application of the conjugation method previously described for the structurally related cytokine rhG-CSF (Ralph et al., 1995). Purity of the DTPA rhIL-2 conjugate, isolated by cation-exchange FPLC, and chelation of 111In were revealed by cation-exchange HPLC. Purity of the conjugate as well as chelation of radiometal were also demonstrated by SDS-PAGE and TLC, respectively. The stoichiometric molar ratio of DTPA to protein for the conjugate was approximately 1:1 as determined by TLC and mass spectrometry. Localization of the DTPA moiety was resolved by a peptide mapping procedure. The protein retained > 95% secondary structure (alpha helicity) following the conjugation. Addition of metal induced an approximate 22% loss of secondary structure for the conjugate. The in vitro biological activity of the protein was unaffected by the conjugated DTPA, even with chelated metal. Pharmacokinetic analysis of DTPA-conjugated cytokines, following chelated 111In, showed clearance and pharmacokinetic parameter values comparable to those of the corresponding unmodified cytokine. DTPA-conjugated cytokines may prove useful in cytokine research, and furthermore may represent a novel class of molecules for imaging, diagnosing, and/or treatment of malignancies where the cytokine receptor is overexpressed. PMID- 7490452 TI - Isolation of leukocytes infiltrating the islets of Langerhans of diabetes-prone mice for flow cytometric analysis. AB - A simple method was developed for flow cytometric immunofluorescence analysis of cells infiltrating the islets of Langerhans in diabetes-prone rodents. Pancreases were submitted to collagenase P digestion and, in order to minimize collagenase action on leukocytes, islets were isolated before digestion was completed, that is, when exocrine tissue still remained around the islets. Islets, maintained in medium supplemented with sodium azide to prevent modulation of surface markers, were then pressed through a metal sieve and the cell suspension filtered through two different nylon screens. Cells were washed before immunofluorescence staining. Comparative phenotyping of spleen cells submitted to a similar treatment showed that, provided the collagenase P batch was screened, this procedure did not alter leukocyte surface markers and allowed multicolor analysis of the islet infiltrating cells. PMID- 7490453 TI - An improved method to create nitrocellulose particles suitable for the immobilization of antigen and antibody. AB - A method is described for producing 1-3 microns sized particles of nitrocellulose (NC) which are able to absorb protein. Protein is absorbed onto preformed particles made by first dissolving a sheet of nitrocellulose paper in DMSO, and then precipitating it with sodium carbonate/bicarbonate buffer. The efficiency of binding is the same as that of an equivalent sheet of non-processed NC filter paper. Antibodies absorbed onto preformed particles are not exposed to DMSO and carbonate buffer and therefore retain a high antigen binding capacity. Antigen and antibody-absorbed NC preformed particles were used to capture antibody and antigen, respectively. Using lysis buffer, the captured antibodies and antigens were readily released from the NC particles. This makes the latter an appropriate matrix for immunoprecipitation assays either for an antigen or for specific antibody. Antigen-coated NC particles were specifically aggregated ('agglutinated') by specific antibodies and thus can be used in semi-quantitative tests. PMID- 7490454 TI - The use of dedicated peptide libraries permits the discovery of high affinity binding peptides. AB - The motif for peptide binding to monoclonal antibody mAb A16, which is known to be directed against glycoprotein D of Herpes simplex virus type 1, was determined using two dedicated peptide libraries. As a starting point for this study we used an A-16 binding lead sequence, which had previously been obtained from a phage display peptide library (Schellekens et al., 1994). Binding studies with different length variants of this peptide identified a 12mer as a suitable lead compound for our library study. Two incomplete dedicated resin-bound synthetic peptide libraries were generated. Both consisted of 2 x 10(6) 12mers, in which positions were alternately fixed (amino acids identical to the lead sequence) and random. The libraries were screened with mAb A16 and beads with binding peptides were sequenced using Edman degradation. This resulted in a unique peptide binding motif, essentially comprising a 7mer core sequence. Comparison of the sequence of the natural epitope with the binding motif revealed that its sequence was identical to the motif except for one position. Substitution of a methionine in the natural epitope by a tyrosine or a phenylalanine at that position, as dictated by the motif, resulted in a peptide with an affinity for binding to mAb A16 about 50 times higher than that of the natural epitope. Thus, if a lead sequence is available, the use of incomplete, dedicated synthetic peptide libraries provides a fast and powerful tool for the detection of high affinity peptides. PMID- 7490455 TI - Hydrogen peroxide destaining: a new method for removing non-specific stains in nitrocellulose membrane-based dot-ELISA for the detection of trypanosomes in tsetse flies (Glossina spp.). AB - Gut samples prepared from laboratory-reared tsetse flies and applied in dots onto nitrocellulose (NC) membrane were found to stain the membrane with differing coloration and intensity. The stains were, predominantly, either reddish to brown or blackish-brown to black and occasionally greenish to almost colourless, depending on the stage of digestion of the bloodmeal in the fly. NC membrane strips applied with tsetse gut samples from T. brucei infected and uninfected control flies were tested with the standard antigen detection dot enzyme-linked immunoassay (dot-ELISA), using a T. brucei specific monoclonal antibody (MoAb) and horseradish peroxidase goat anti-mouse conjugate. The stains in both infected and uninfected sample dots persisted through the assay. Furthermore, the staining intensity of some assayed uninfected sample dots were enhanced as a result of non specific reactivity, making it difficult to distinguish between the infected and uninfected flies. This necessitated the development of a simple technique by which the non-specific stains and reactions could be removed. Sample 'dotted' NC membrane strips were destained by incubation with 5% hydrogen peroxide (H2O2) diluted in 5% skimmed milk in Tris buffer, pH 8.0. After washing, the destained strips were tested in the dot-ELISA. This method gave satisfactory reproducible results, since the most intense stains could be removed, and it had no effect on trypanosome antigens detected by a panel of four T. brucei species-specific, three T. vivax species-specific, four T. congolense species-specific and four Nannomonas subgenus-specific MoAbs. Using the destaining process in a modified dot-ELISA, 86 out of 95 (90.5%) of Glossina morsitans centralis flies experimentally infected with T. brucei, were identified. The destaining method was also used successfully to decolorize NC membrane bound tsetse faecal material. PMID- 7490456 TI - Determination of tissue inhibitor of metalloproteinases-2 (TIMP-2) in experimental animals using monoclonal antibodies against TIMP-2-specific oligopeptides. AB - We have developed a one-step sandwich enzyme immunoassay (EIA) using monoclonal antibodies against oligopeptides of the human tissue inhibitor of metalloproteinases-2 (TIMP-2) (Fujimoto et al. (1993) Clin. Chim. Acta 220, 31). The present studies further demonstrated that the antibodies cross-react with TIMP-2 species of experimental animals including mouse, rat, guinea pig and rabbit. The detection of the TIMP-2 species in our EIA system was verified using rat TIMP-2 and the EIA was subsequently used to measure the animal TIMP-2 in the sera. Using human TIMP-2 as a standard, TIMP-2 levels in the sera of mouse, rat, guinea pig and rabbit were approximately 80, 200, 270 and 25 ng/ml, respectively. PMID- 7490457 TI - Characterization of mouse nasal lymphocytes isolated by enzymatic extraction with collagenase. AB - A reproducible method for isolation of mouse nasal lymphocytes was developed. The cells were released from tissue fragments of dissected mouse nose by enzyme extraction with collagenase and separated by a stepwise Percoll gradient centrifugation. The partially purified nasal lymphocyte fraction from normal BALB/c mice contained CD4+ T cells (18-23%), CD8+ T cells (7-10%) and B cells (20 38%), when analysed with a FACScan fluorescence analyser. The ratio of T to B cells and that of CD4+ to CD8+ T cells in the nasal cell fraction were about twice as high as those in Peyer's path lymphocytes. The nasal lymphocyte fraction from the mice infected with influenza virus was then characterized. The nasal lymphocytes contained a twice larger number of CD4+ and a three times larger number of CD8+ T cells than those of normal mice 7 days after infection. They produced IFN-gamma which increased after infection. They contained the cells secreting influenza virus-specific IgA and IgG antibodies 4 weeks after infection. Moreover, the nasal lymphocytes from infected C3H mice lysed the virus infected-target cells. These results suggest that this method can successfully be used for investigating cellular dynamics of mucosal immunity in the upper respiratory tract of experimental animals. PMID- 7490458 TI - Fluorescence resonance energy transfer as a new method for the epitope-specific characterization of anti-platelet antibodies. AB - The detection and characterization of anti-platelet antibodies which are directed to HLA class I molecules or platelet-specific glycoproteins is of great importance in the diagnosis and treatment of thrombocytopenia. In this paper a simple and rapid flow cytometric assay for the epitope-specific characterization of anti-platelet antibodies is described using fluorescence resonance energy transfer (FRET). Patient platelets or test platelets preincubated with patient serum were analyzed for surface-bound immunoglobulins using R-phycoerythrin conjugated polyclonal anti-human IgG antibodies (excitation 488 nm, emission 585 nm). In a second step, HLA class I structures, platelet-specific glycoproteins (gpIIb/IIIa, gpIb), and the Fc gamma receptor II were stained with murine monoclonal antibodies and Cyan 5-labelled polyclonal anti-mouse IgG antibodies (excitation 585 nm, emission 670 nm). Upon monochromatic fluorescence excitation with a 488 nm argon laser the efficiency of light transfer from R-phycoerythrin to Cyan 5 is a direct measure of the distance between the human platelet-bound antibody and the epitope detected by the murine monoclonal antibody (mab). The assay permits discrimination between human antibodies directed to different platelet-specific glycoproteins or HLA class I structures without interference from non-specific Fc gamma receptor-bound immune complexes and also between antibodies directed to different epitopes on glycoprotein heterodimers (e.g., gpIIb/IIIa). PMID- 7490459 TI - Fluorescence labeling of bacteria for studies of intracellular pathogenesis. AB - Interactions between intracellular bacterial pathogens and their eukaryotic cellular hosts or targets are often studied with fluorescence-based techniques such as fluorescence microscopy and flow cytometry. We tested whether the intracellular bacterial pathogens L. monocytogenes, M. avium, M. tuberculosis, and S. typhimurium could be labeled by growth in broth containing the fluorochromes carboxy-X-rhodamine (CR), a hydrazine derivative of fluorescein (FH), and Lucifer Yellow CH (LY). Only Listeria were labeled by all three fluorochromes, Salmonella took up only FH, whereas M. avium and M. tuberculosis were labeled by FH and LY. In general, the fluorochromes did not affect bacterial growth or viability, although FH inhibited the growth of M. tuberculosis. Fluorescent Listeria and M. tuberculosis were used to demonstrate that FH- and LY labeled bacteria were bright enough after phagocytosis by macrophages to distinguish phagocytic from nonphagocytic cells by flow cytometry. To test whether the fluorochromes might alter bacterial interactions with host cells, we measured both phagocytosis of fluorescent Listeria by macrophages and subsequent bacterial replication in these cells. In these experiments, labeled and unlabeled Listeria were phagocytosed similarly by macrophages and were not impaired in their ability to replicate within them. Thus, this method for fluorescence labeling of bacteria is useful for studying physiologic macrophage:bacteria interactions. PMID- 7490460 TI - Purification of a 92 kDa human immunostimulating glycoprotein obtained from the Tamm-Horsfall glycoprotein. AB - A purification method for a human urinary glycoprotein (HGP92) dissociated from Tamm Horsfall protein (THP) is described. Tamm-Horsfall protein, obtained by salt precipitations, was again precipitated in presence of monovalent ions. In these conditions, Tamm-Horsfall protein displayed a tendency to form a gel. After ultracentrifugation, HGP92, which was trapped in the gel, was dissociated from Tamm-Horsfall protein and found in the supernatant. The final step of purification of HGP92 was chromatography on a DEAE Affigel blue column. Injected intravenously, HGP92, but not THP, protected mice against a lethal inoculum of Listeria monocytogenes. This procedure has the advantage of being easy to perform, and enables preparation of large amounts of HGP92. These results suggest that the previously described 'immunostimulating' properties of Tamm-Horsfall protein were, in fact, a consequence of its contamination by HGP92. PMID- 7490461 TI - Comparison of [3H]thymidine incorporation with MTT- and MTS-based bioassays for human and murine IL-2 and IL-4 analysis. Tetrazolium assays provide markedly enhanced sensitivity. AB - The sensitivity of [3H]thymidine incorporation and MTT/MTS colorimetric bioassays for detection and quantitation of murine and human IL-4 and IL-2 were compared in CT.4S, CT.h4S and HT-2 bioassays respectively. We reasoned that low levels of cytokine, insufficient to induce cell proliferation (thus, DNA synthesis and [3H]thymidine incorporation), may be sufficient to maintain the viability of the bioassay cells in culture. Because colorimetric assays such as those employing MTT (3-(4,5 dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide) or MTS (3 (4,5-dimethylthiazol-2-yl)-5-(3-carboxymethoxyphenyl)-2-( 4-sulfonyl)-2H- tetrazolium) measure conversion of these salts to intensely colored formazan products by mitochondrial enzymes independent of whether proliferation is induced, we reasoned that such assays could be superior for detection of low levels of cytokine protein. Direct comparison of these approaches demonstrated that the MTT- and MTS-based assays were consistently able to detect 2-16-fold lower cytokine levels than methods based on [3H]thymidine incorporation. Moreover, the MTT and MTS assays exhibited higher precision with standard deviations of < 1-4% vs. 5-15% for thymidine incorporation. This finding is of particular importance in approaches such as limiting dilution analysis, or primary bulk culture of antigen-stimulated human lymphocytes, where levels of cytokine production may be extremely low. PMID- 7490463 TI - An assay for the identification of antigens recognized by cytotoxic T cells, based on transient transfection of COS cells. AB - The studies reported here describe methodology permitting the direct identification of antigens recognized by cytotoxic T lymphocytes. We demonstrated that bovine alloreactive CTL can detect a bovine MHC molecule transiently expressed in a COS cell population in a standard microcytotoxicity assay. We then showed that alloreactive CTL can detect cells expressing the bovine class I MHC molecule in a population of cells transfected with the plasmid containing the corresponding gene plus 100-fold as many plasmids containing an irrelevant gene. In addition, the transiently transfected COS cells can specifically restimulate CTL as detected by a standard microcytotoxicity assay using the target cell line. Overall, the results suggest that COS cells could be employed for the direct screening of an antigen or antigen gene library by immune CTL. PMID- 7490464 TI - Elevated numbers of proliferating mononuclear cells in the peripheral blood of psoriatic patients correlate with disease severity. AB - Although psoriasis is characterized by the accumulation of activated proliferating lymphoid cells in the psoriatic skin lesion, it is not known whether these cells are activated and proliferating before entry into the psoriatic plaque. The current study evaluates the number and phenotype of proliferating lymphoid cells in the blood of psoriatic patients. Proliferation of peripheral blood mononuclear cells was evaluated on cytospun preparations of these cells using autoradiographic techniques after pulsing the mononuclear cells with 3H-methyl thymidine for 2 h. The phenotypes of the labeled peripheral blood mononuclear cells were determined combining autoradiography and immunohistochemistry with monoclonal antibodies directed at CD3, CD4, CD8, CD11c, CD22, and human leukocyte antigen-DR. The data demonstrated elevated numbers of proliferating lymphoid cells in the blood of psoriatic patients compared with normal nonpsoriatic volunteers (p < 0.01). Furthermore, the number of circulating proliferating mononuclear cells increased significantly with increasing psoriasis skin disease severity (correlation coefficient 0.95; p < 0.0001). When the phenotype of the proliferating cells in the blood was examined, the numbers of T cells (CD3+, CD4+, CD8+ cells), B cells (CD22+ cells), monocytes (CD11c+ cells), and human leukocyte antigen-DR+ cells were significantly elevated compared with nonpsoriatic skin (p < 0.01) and increased with increasing disease activity (correlation coefficient range 0.48-0.74; p < 0.05). The data suggest a generalized systemic activation of T, B, and monocytic cells that results in labeling of up to 0.16% of the circulating mononuclear cells with 3H-methyl thymidine (i.e., proliferating and presumably activated) when assayed in vitro. PMID- 7490465 TI - Low doses of repetitive ultraviolet A induce morphologic changes in human skin. AB - Repetitive exposure of skin to sunlight is known to result in dermatoheliosis, characterized by photoaging and carcinogenesis. It has been demonstrated previously that relatively large amounts of ultraviolet (UV) A can produce photodamage and it is believed that UVB plays a major role in the induction of photodamage and photocarcinogenesis. The study reported here determines the cutaneous effects of minimal erythemal amounts of solar-simulated UV radiation as well as suberythemal and minimal erythemal doses of UVA. Previously non sunexposed human skin was irradiated twice weekly for 24 weeks. Biopsies were obtained 12, 24 and 36 weeks after the initial irradiation and assessed for both epidermal and dermal alterations. Dermal elastic tissue content was measured via computerized image analysis. All UV treatment regimens produced observable epidermal and dermal changes. These alterations were observed after only 12 weeks of twice-weekly irradiation and were still evident 12 weeks after the final irradiation. Interestingly, UVA irradiation produced a decrease in elastic tissue content whereas solar-simulated UV produced a slight increase. Most notable were the changes produced by the suberythemal dose of UVA. Surprisingly, this relatively low UVA dose produced a reduction in elastic tissue content. The results of this investigation demonstrate that small amounts of UVA or solar simulated UV are capable of producing cutaneous photodamage. These findings suggest that even suberythemal doses of repetitive UVA may lead to photoaging of the skin and that there is a need for daily broad spectrum UV protection. PMID- 7490462 TI - Analysis of T cell receptor beta chain expression by isoelectric focusing following gene amplification and in vitro translation. AB - We describe a new approach to analysis of T cell receptor diversity based on isoelectric focusing of in vitro translation products of amplified V region genes. The method is illustrated by analysis of V beta 2 profiles in peripheral blood lymphocytes from normal donors. The primers used for V beta 2 analysis spanned the V-(D-)J junction and included the segment from amino acid residue position 53 in the variable region to residue 132 of the constant region. The isoelectric focusing patterns display approximately 13-14 bands of varying intensity. Differences in expression of V beta 2-derived peptides were detected in comparisons of the isoelectric focusing profiles from different individuals, suggesting that the method may be useful for detecting genetically determined, immune response related or disease associated differences in Tcr V region expression. The major isoelectric focusing bands have been interpreted as representing groups of V beta 2 sequences sharing J beta region and NDN region charge similarity. Quantitative differences were detected in V beta 2 profiles of CD4 and CD8 T cell subpopulations indicating there may be selection for different charge characteristics in NDNJ sequences in the two T cell subsets. The method provides a new dimension for the detection of perturbations in the T cell repertoire. PMID- 7490466 TI - Endothelin-1 of keratinocyte origin is a mediator of melanocyte dendricity. AB - Melanocytes synthesize melanin and transfer it to keratinocytes via dendritic processes. Keratinocytes are known to produce constitutively several factors, including endothelin-1 (ET-1), that together affect melanocyte proliferation, migration, melanogenesis, and dendrite formation. After ultraviolet (UV) irradiation, synthesis and secretion of ET-1 are up-regulated in keratinocytes. Because UV irradiation of skin is known to be associated with increased melanocyte dendricity, and because medium conditioned by UV-irradiated keratinocytes (UV-KCM) induces melanocyte dendricity to a greater degree than does baseline keratinocyte-conditioned medium (KCM), we investigated whether ET-1 promotes melanocyte dendricity. ET-1, originally recognized as a vasoconstrictive peptide, has recently been shown to stimulate melanocyte proliferation and tyrosinase activity. We now report that ET-1 supplementation of cultured melanocytes significantly increases the percentage of dendritic melanocytes, as well as dendrite length, in a dose-dependent manner. Moreover, UV-KCM was found to contain over 25-fold more ET-1 than KCM, and ET-1 supplementation of KCM induced melanocyte dendricity comparable to that induced by UV-KCM. Further, melanocyte dendricity induced by UV-KCM was significantly inhibited by the addition of anti-ET-1 monoclonal antibody to the medium, suggesting that the UV KCM effect on melanocyte dendricity is mediated largely through ET-1. Our findings suggest that in the skin, ET-1 of keratinocyte origin promotes melanocyte dendricity in response to UV irradiation. PMID- 7490467 TI - Immediate-type hypersensitivity response followed by a late reaction is induced by repeated epicutaneous application of contact sensitizing agents in mice. AB - Repeated administration of antigen often leads to consequences different from those expected with fewer encounters with the antigen, but little attention has been paid to the effects of repeated epicutaneous application of antigens. To investigate whether repeated epicutaneous application of a contact-sensitizing agent that is generally thought to evoke a typical delayed-type hypersensitivity response could result in adverse or different consequences, BALB/c mice were sensitized with 2,4,6-trinitro-1-chlorobenzine and then were repeatedly elicited on the original sensitized site with the same antigen for 24-48 d. Detailed analyses showed that the time-course of antigen-specific hypersensitivity responses shifted from a delayed-type hypersensitivity to an immediate-type response followed by a late reaction as epicutaneous applications were repeated, a finding different from that previously reported. Development of these hypersensitivity responses was antigen specific, and this shift was associated with epidermal hyperplasia, accumulation of large numbers of mast cells and CD4+ T cells beneath the epidermis, and elevated serum levels of antigen-specific IgE. The immediate-type response to 2,4,6-trinitro-1-chlorobenzine was also induced in 2,4,6-trinitro-1-chlorobenzine-treated, genetically mast cell-deficient W/Wv mice that contained significant numbers of mast cells, but not in similarly treated S1/S1d mice devoid of mast cells. Our experimental system would provide a simple, reproducible animal model for chronic skin inflammation induced by various antigens. PMID- 7490469 TI - MAT-1, a monoclonal antibody that specifically recognizes human tyrosinase. AB - There is no available monoclonal antibody which reacts specifically recognizes human tyrosinase. Employing a synthetic peptide, MEKEDYHSLYQSHL, corresponding to the carboxyl terminus of human tyrosinase as an immunogen, we produced a mouse monoclonal antibody MAT-1 of the IgG1 isotype. The epitope for MAT-1 was determined to be EDYH, the sequence of which is not present in human tyrosinase related protein-1 (TRP-1) or tyrosinase-related protein-2 (TRP-2). By transient expression assays and immunofluorescence technique, we show that MAT-1 reacts specifically with cells expressing human tyrosinase cDNA but not with cells expressing TRP-1 or TRP-2 cDNA. The results of immunohistochemical staining also confirmed that MAT-1 reacts specifically with epidermal melanocytes in human skin sections. MAT-1 should be invaluable for studying the interaction between tyrosinase and TRPs and for detecting the changes in the levels of tyrosinase expression. In addition, MAT-1 should be useful as a sensitive immunohistochemical tool for investigation of various pigmentary disorders and possibly for the diagnosis of melanoma. PMID- 7490468 TI - Genetically modified human epidermis overexpressing PDGF-A directs the development of a cellular and vascular connective tissue stroma when transplanted to athymic mice--implications for the use of genetically modified keratinocytes to modulate dermal regeneration. AB - We investigated the hypothesis that keratinocyte-produced platelet-derived growth factor-AA (PDGF-AA) is involved in epidermal-dermal interactions and that PDGF-AA is an important mediator of the temporal and spatial events of tissue repair. Retroviral-mediated gene transfer was used to introduce the gene encoding human PDGF-A into cultures of human diploid keratinocytes. Genetic modification boosted the endogenous in vitro level of PDGF-AA secretion by over 300 fold. When PDGF secreting cells were transplanted as epithelial sheets to athymic mice, modified keratinocytes underwent terminal differentiation and generated a stratified epithelium comparable to unmodified cells. Seven days after grafting the newly synthesized connective tissue layer subjacent to the PDGF-A-modified grafts was significantly thicker, was rich in mononuclear cells and fibroblasts, and had increased numbers of blood vessels when compared to control grafts of unmodified cells. These results suggest that PDGF-AA secreted by the epidermis is an important mediator of epithelial-mesenchymal interactions and helps to promote growth and vascularization of the underlying dermal tissue. Further, these data demonstrate the feasibility of using genetically modified cells to modulate tissue regeneration. PMID- 7490470 TI - Maintenance of donor phenotype after full-thickness skin transplantation from mice with chronic proliferative dermatitis (cpdm/cpdm) to C57BL/Ka and nude mice and vice versa. AB - Chronic proliferative dermatitis is a spontaneous mutation in C57BL/Ka mice (cpdm/cpdm) and is characterized by epithelial hyperproliferation, infiltration by eosinophils and macrophages, and vascular dilatation. To elucidate whether these pathologic features are the result of a local (skin) process or a consequence of a systemic disorder, transplantations were performed of full thickness grafts of affected skin from cpdm/cpdm mice and normal skin from control (C57BL/Ka) mice on the back of cpdm/cpdm, C57BL/Ka and athymic nude mice. After 3 months, the grafts maintained the histologic phenotype of the donor animal. Intercellular adhesion molecule-1 continued to be expressed by basal keratinocytes of the cpdm/cpdm grafts after transplantation. In contrast, the basal keratinocytes of the C57BL/Ka grafts onto cpdm/cpdm mice remained negative for intercellular adhesion molecule-1 3 months after transplantation. An increased number of proliferating keratinocytes was present in the cpdm/cpdm skin graft transplanted to nudes or to C57BL/Ka mice based on short-term bromodeoxyuridine labeling. The bromodeoxyuridine incorporation in the keratinocytes of the control C57BL/Ka skin grafts transplanted to cpdm/cpdm, nude, or C57BL/Ka mice was the same as in the keratinocytes of normal C57BL/Ka mice. This study demonstrates that the pathologic features found in the cpdm/cpdm mice are the result of a disorder in the epidermis or dermis and not due to a systemic defect. PMID- 7490472 TI - In human dermis, ultraviolet radiation induces expansion of a CD36+ CD11b+ CD1- macrophage subset by infiltration and proliferation; CD1+ Langerhans-like dendritic antigen-presenting cells are concomitantly depleted. AB - Antigen-presenting (APC), suppressor T-cell-inducing macrophages infiltrate both human and murine epidermis after ultraviolet radiation (UVR) exposure. To determine their derivation, we prepared epidermal cell and dermal cell suspensions from human keratome biopsy specimens obtained from nonexposed skin and from UVB-irradiated sites (3 d after four times the minimal erythema dose). Simultaneous triple-marker flow cytometric analysis established the extended phenotype of macrophages infiltrating sunburned human epidermis (CD1a- CD1c- CD11b+ CD11c+ CD36+ Fc gamma RII+ DR+). This then enabled us to track dermal cells of this phenotype after UVR in relation to the heterogeneous DR+ populations in normal dermis. By both in situ immunohistology and cell suspension flow cytometry, UVR induced an expansion of bone marrow-derived DR+ cells in the perivasculature and sub-basement membrane zone of the papillary dermis. Despite an overall expansion of DR+ cells, the CD1a+ CD1c+ CD36- DR+ Langerhans-cell-like dendritic APC subset of dermal DR+ cells was depleted (p < 0.05), indicating that UVR-induced epidermal Langerhans cell loss (from 95% to 7% of DR+ epidermal cells) is not accounted for by Langerhans cell accumulation in the dermis. By contrast, UVR exposure induced a selective expansion of the dermal macrophage subset, which is phenotypically identical to the monocytic/macrophagic APCs that appear in the epidermis after UV injury (p < 0.01). Cell cycle analysis (to determine whether this expansion was accounted for entirely by infiltration) revealed no increase in the percentage of DR+ CD36+ UVR-exposed dermal cells in S/G2/M phase; however, the expanded DR+ CD36+ subset continued its already substantial level of proliferation unabated. Therefore, epidermal macrophages derive not only from transcapillary migration, but also from in situ proliferation of a dermal precursor. Taken together, these findings show that UVR creates an epidermal and dermal APC milieu which is dominated by monocytic/macrophagic cells, through depletion of cells of dentritic APC phenotype, and concomitant selective dermal expansion of a CD1a- CD1c- CD11b+ CD36+ Fc gamma RII+ DR+ (monocyte/macrophage) population. PMID- 7490471 TI - A nicotinic acetylcholine receptor regulating cell adhesion and motility is expressed in human keratinocytes. AB - Acetylcholine is synthesized and released by human epidermal keratinocytes and modulates the adhesion and motility of these cells. To understand the molecular basis of the effects of acetylcholine on keratinocytes, we investigated the presence, pharmacology, structure, and function of nicotinic acetylcholine receptors in human epidermal keratinocytes. Patch-clamp studies indicated that keratinocytes express acetylcholine receptors with ion gating and pharmacologic properties similar to those observed so far only in neurons, and containing the alpha 3 subunit. Specific binding of the receptor-specific ligand 125I-kappa bungarotoxin revealed approximately 5500 binding sites per cell on undifferentiated keratinocytes in cell cultures and approximately 35,400 binding sites per cell on mature keratinocytes freshly isolated from human neonatal foreskins. Antibody binding and polymerase chain reaction experiments demonstrated the presence of alpha 3, beta 2, and beta 4 nicotinic receptor subunits. Binding of subunit-specific antibodies indicated that nicotinic receptors were associated with the suprabasal keratinocytes in epidermis and localized to the cell membranes of differentiated keratinocytes in cell cultures. Acetylcholine and the nicotinic agonist nicotine increased cell-substrate and cell-cell adherence of cultured keratinocytes and stimulated their lateral migration. The specific antagonists kappa-bungarotoxin and mecamylamine caused cell detachment and abolished migration. Thus, a nicotinic receptor expressed in keratinocytes may mediate acetylcholine control of keratinocyte adhesion and motility. PMID- 7490473 TI - Mast cells induce T-cell adhesion to human fibroblasts by regulating intercellular adhesion molecule-1 and vascular cell adhesion molecule-1 expression. AB - The capacity of mast cell products to mediate T-cell adhesion to fibroblasts was explored using heterotypic coculture systems or by exposing fibroblasts to mast cell-conditioned media (MCCM), prepared by degranulating mast cells with calcium ionophore. Experimental results indicated that fibroblasts exposed to MCCM for 24 h bound fivefold more T cells than control fibroblasts. Binding was inhibited with intercellular adhesion molecule-1 (ICAM-1) or vascular cell adhesion molecule-1 (VCAM-1) neutralizing antibodies. Enzyme-linked immunosorbent assay and fluorescence-activated cell sorter analysis revealed that fibroblasts exposed to MCCM markedly increased ICAM-1 and VCAM-1 surface expression by 4 h, with levels maximal at 16 h and returning toward baseline by 48 h. A dose-dependent response of ICAM-1 and VCAM-1 expression was noted using serial dilutions of MCCM or by altering the ratio of degranulated mast cells cocultured with fibroblasts. Similar results were obtained using human fibroblasts derived from the dermis, synovium, and lung, although lung fibroblasts were generally less responsive. Northern analysis confirmed that MCCM regulated ICAM-1 and VCAM-1 expression at the mRNA level. In summary, mast cell products stimulated fibroblast surface expression, steady-state mRNA levels, and functional expression of ICAM-1 and VCAM-1. Experimental data suggest that mast-cell-derived tumor necrosis factor alpha may be in large part responsible for these observations, although further studies using human mast cells will be required. Using a skin-equivalent organotypic coculture model with fibroblasts admixed with mast cells, we observed increased ICAM-1 expression in both keratinocytes and fibroblasts after activation of the mast cells. PMID- 7490474 TI - Computer simulations of histologic patterns in melanoma using a cellular automaton provide correlations with prognosis. AB - Computer simulations have been used frequently in the life sciences to investigate the mechanisms of morphologic pattern formation. The cellular automaton program SMN5 is designed to simulate tumor growth and to estimate biologic properties by comparing real tumor patterns with computer-simulated reference patterns. This method was applied to 195 cases of primary melanoma of the skin. S-100-stained sections were evaluated by image analysis and compared statistically to a reference set of 4000 simulated patterns. Estimates of tumor cell proliferation, motility, cell loss, cohesion, stroma destruction, and intercellular signals (autocrine and paracrine factors affecting growth, motility, and cell loss) were calculated. Twelve of 18 estimated parameters correlated significantly with tumor progression, as indicated by vertical tumor thickness (linear regression analysis: p < or = 0.05), and 13 of 18 parameters carried prognostic significance (log rank test: p < or = 0.05). Poor prognosis was associated particularly with a pronounced increase in the estimates of proliferation, tumor cell motility, and stromal degradation. Poor prognosis was also associated with a decrease in the estimates of cell loss, tumor cell cohesion, and paracrine growth factor dependence. In multivariate analysis using Cox's proportional hazard model, stromal degradation and motility showed prognostic information in addition to conventional prognostic parameters. The study shows that analytical comparison of real tumors with computer-simulated patterns of a cellular automaton facilitates a functional interpretation of tumor morphology, which carries prognostic significance in cutaneous melanoma. PMID- 7490475 TI - Expression of amphiregulin is regulated in cultured human keratinocytes and in developing fetal skin. AB - Previous studies have indicated that amphiregulin is a major autocrine factor for human keratinocytes. To evaluate the possibilities that amphiregulin could function in fetal skin morphogenesis and contribute to the growth regulation of epidermis, immunostaining with a specific anti-amphiregulin monoclonal antibody was observed at different stages of fetal skin development, and the results were compared with neonatal and adult skin specimens and cultured neonatal keratinocytes. Immunoreactive amphiregulin was readily detected in the periderm and basal epidermal layers of embryonic epidermis but became gradually less detectable in the periderm concurrent with an increase in staining of the spinous layer as it developed during the fetal period. Basal and spinous keratinocyte expression of amphiregulin was predominantly cytoplasmic, but with punctate nuclear foci, and this pattern persisted into the neonatal period. At all developmental stages, epithelial and mesenchymal cells of the follicle were reactive, often in a nuclear pattern. Dermal mesenchymal cells were increasingly reactive in late fetal skin, but the staining decreased postnatally. In adult skin only randomly scattered nuclei of spinous keratinocytes and follicular structures such as the inner root sheath were stained. Examination by scanning laser confocal microscopy of cultured neonatal keratinocytes showed a nonrandom distribution of amphiregulin to the peripheral cytoplasm and plasma membranes at the outer perimeter of cell colonies, with much less reactivity of apposed keratinocyte membranes at interior sites. Nuclei were heterogeneously stained. Amphiregulin reactivity declined at higher cell densities. These data indicate that expression of amphiregulin is regulated in vitro and developmentally during cutaneous morphogenesis. PMID- 7490476 TI - Interferon-gamma-dependent stimulation of Fas antigen in SV40-transformed human keratinocytes: modulation of the apoptotic process by protein kinase C. AB - Fas antigen is a cell membrane protein that has been suggested to mediate apoptosis. Using SV40-transformed human keratinocytes, we investigated the Fas antigen-dependent apoptotic process. The expression of Fas antigen mRNA was markedly induced by interferon-gamma (IFN-gamma) treatment (500 U/ml). After IFN gamma treatment in the presence of anti-Fas monoclonal antibody, apoptosis was induced, as detected by the formation of nucleosome-sized fragments of DNA and morphologically by apoptotic cells with round homogeneous nuclear beads detected by acridine orange staining. The apoptotic SV40-transformed keratinocytes were analyzed quantitatively by enzyme-linked immunosorbent assay using antihistone and peroxidase-conjugated anti-DNA antibodies to detect cell death. The IFN-gamma and anti-Fas antibody-dependent apoptotis was observed by 3 h, and the maximal response was observed by 12 h. The induction of apoptosis was significantly augmented by treatment with 10 ng/ml 12-o-tetradecanoyl-phorbol-13-acetate (TPA). TPA alone had no effect on either Fas antigen expression or on the apoptotic process. Other protein kinase C activators (1-oleoyl-2-acetylglycerol and mezerein) also stimulated IFN-gamma-dependent apoptosis, whereas 4-o-methyl phorbol myristate acetate, a very weak protein kinase C activator, had only a slight effect. The TPA-induced augmentation of apoptosis was inhibited by the protein kinase C inhibitor, 1-(5-isoquinoline-sulfonyl)-2-methyl piperazine dihydrochloride (H-7). However, H-7 inhibited only the TPA-induced augmentation of apoptosis; the basal IFN-gamma- and anti-Fas-dependent apoptosis remained in the presence of H-7. Northern blot analysis revealed that c-jun mRNA was induced by IFN-gamma plus anti-Fas antibody treatment as well as by TPA treatment; the addition of IFN-gamma alone to the incubation medium had no effect on the expression of c-jun mRNA. These results indicate that IFN-gamma induces a Fas antigen-dependent apoptotic process in SV40-transformed keratinocytes and that TPA augments the process through the activation of protein kinase C. PMID- 7490477 TI - Identification of functional platelet-activating factor receptors on human keratinocytes. AB - Platelet-activating factor (PAF) is a potent inflammatory mediator that has been shown to be produced by human keratinocytes and is thought to play a role in cutaneous inflammation. Immunofluorescence and radioligand binding studies were used to characterize PAF receptors (PAF-R) on human keratinocytes and the human epidermoid cell lines A-431 and HaCaT. Indirect immunofluorescence studies demonstrated anti-PAF-R staining of primary cultures of human keratinocytes, A 431 cells, and HaCaT cells. Primary cultures of human fibroblasts and the melanoma cell line SK-30 failed to show immunostaining above that seen with control antiserum. With indirect immunofluorescence studies of sections of normal human skin, a granular anti-PAF-R staining pattern was noted on the keratinocyte cell membranes. A-431 cells readily metabolized PAF by deacetylation-reacylation at 37 degrees C, but not at 4 degrees C. Binding studies on crude membrane preparations of A-431 cells conducted at 4 degrees C demonstrated specific binding that reached saturation by 120 min. Scatchard analysis of PAF binding data revealed a single class of high-affinity (KD = 6.3 +/- 0.3 nM) PAF binding sites. The immunofluorescence and radioligand binding sites were shown to be functional PAF-Rs, as 10 pM to 1 microM PAF increased intracellular calcium in primary cultures of human keratinocytes, A-431 cells, and HaCaT cells, whereas PAF treatment of primary cultures of human fibroblasts or the melanoma cell line SK-30 did not result in changes in the intracellular calcium concentration. The structurally dissimilar PAF-R antagonists CV-6209, Ro19-3704, and alprazolam all inhibited the PAF-induced calcium changes in A-431 cells. The CV-6209 inhibition was seen at doses that competed with the PAF binding to these cells. These studies provide the first evidence for the presence of a functional PAF-R expressed on human keratinocytes, suggesting that this lipid mediator may play an important role in normal keratinocytes or in inflammatory dermatology. PMID- 7490478 TI - In situ-bound antibodies eluted from the skin of patients with bullous pemphigoid are preferentially directed against the 230-kD bullous pemphigoid antigen. AB - Two bullous pemphigoid antigens (BPAs) have been identified: a 230-kD intracellular hemidesmosome-associated molecule and a 180-kD transmembrane hemidesmosome-associated molecule. Although patients with bullous pemphigoid (BP) have been shown to have circulating antibodies directed against one or both BPAs, the antigenic specificity of tissue-bound BP autoantibodies has not been studied. Because these skin-bound antibodies may play an important role in disease initiation, we sought to determine their antigenic specificity. In situ-bound BP antibodies were eluted with glycine from salt-split perilesional skin biopsy specimens and subjected to immunoprecipitation. Ten of 13 patients had glycine eluted antibodies that recognized the 230-kD BPA, whereas four of 13 patients had glycine-eluted antibodies that recognized the 180-kD BPA. When these glycine eluted BP skin specimens were sequentially eluted with urea, we found either antibodies of the same specificity or loss of reactivity to one of the BPAs, but no reactivity to any BPAs previously undetected with glycine elution. Sequential elution with sodium dodecylsulfate revealed no detectable BP antibodies. The heavy- and light-chain isotypes of the circulating, tissue-bound, and eluted BP antibodies were very similar, suggesting that we are not eluting certain subsets of BP antibodies. Sera from these patients contained circulating antibodies that recognized either one or both BPAs. Our observations demonstrate that in situ bound antibodies eluted from the skin of patients with BP are preferentially directed against the 230-kD BPA. PMID- 7490479 TI - Phenotypic and functional heterogeneity among murine epidermal-derived dendritic cell clones. AB - We have established recently long-term dendritic cell lines from the epidermis of newborn BALB/c mice. These lines, termed XS series, resembled epidermal resident Langerhans cells or their progenitors in terms of surface phenotype, antigen presenting capacity, and growth factor requirement. We examined in this study the degree of clonal heterogeneity among XS cells with respect to each of these features. Twelve stable clones were established by limiting dilution microculture from 8-10-week-old cultures of the XS52 or XS20 line. Despite the uniform expression of CD45, these clones varied substantially in their expression of Ia, B7-1, and B7-2 molecules. They also varied significantly in their relative efficiency in activating T cells. Finally, remarkable clone-to-clone heterogeneity was also observed in their growth factor responsiveness; some clones responded equally well to granulocyte macrophage-colony-stimulating factor 1, whereas others responded preferentially to one or the other of these factors. We propose that the observed clonal heterogeneity in XS cells reflects possible heterogeneity in the state of maturation and mitotic potential among the starting populations, i.e., skin-associated dendritic cells in newborn mice. PMID- 7490480 TI - Interleukin (IL)-15 promotes the growth of murine epidermal gamma delta T cells by a mechanism involving the beta- and gamma c-chains of the IL-2 receptor. AB - Dendritic epidermal T cells (DETC) are skin-specific members of the epithelial gamma delta T-cell family in mice. We have reported previously that the growth of DETC is promoted by interleukin (IL)-2 in an autocrine fashion, or by IL-7, which is secreted by neighboring keratinocytes. Here we report that DETC growth is promoted by IL-15, a newly discovered T-cell growth factor that is produced in lymphoid as well as nonlymphoid tissues. Recombinant IL-15 promoted the growth of the 7-17 DETC line in a time- and dose-dependent fashion. Using monoclonal antibodies against alpha-, beta-, or gamma c-chains of the IL-2 receptor complex, we observed that the combination of anti-beta chain and anti-gamma c chain antibodies blocked IL-15 responsiveness completely, whereas anti-alpha chain had no effect. These results indicate that this gamma delta T-cell line uses the beta/gamma c heterodimer for proliferative responses to IL-15. Antibodies against IL-2 or IL-7 did not block IL-15-driven proliferation of 7-17 DETC, indicating that IL-15 promotes their growth in an IL-2- and IL-7-independent manner. Both the surface expression of beta/gamma c heterodimers and the IL-15 responsiveness of 7-17 DETC were highest 1 to 8 days after concanavalin A stimulation, and both declined substantially 21 days after stimulation, illustrating regulation by the state of cell activation. Working with epidermal cells that were freshly procured from CBA mice, we noted that IL-15 promoted conavalin-A-triggered growth of Thy 1+ cells (i.e., DETC), but not of the Thy-1- cells. The gamma c-chain was not expressed by freshly procured DETC, becoming detectable within 48 h after concanavalin A stimulation. We propose that IL-15 facilitates the growth of epithelial gamma delta T cells by a beta/gamma c receptor-dependent mechanism. PMID- 7490481 TI - Characterization of hair follicle bulge in human fetal skin: the human fetal bulge is a pool of undifferentiated keratinocytes. AB - It has been suggested that the bulge of the hair follicle contains a pool of follicular stem cells that may serve as a target site of graft-versus-host disease and as a source of cells with carcinogenic potential. The bulge is prominent in the developing follicle although it is a subtle swelling in the adult follicle. In this paper, we studied the bulge in human fetal skin specimens. Ultrastructurally, the bulge cells, especially the interior cells, have abundant free ribosomes and glycogen particles, but almost no cytoplasmic organelles indicative of differentiation. Immunostaining with several specific anti-keratin antibodies demonstrated that the bulge cells express keratins of both stratified and simple epithelia. Melanocytes and Merkel cells, defined by immunohistochemical and ultrastructural criteria, are seen among bulge cells. Laser confocal microscopy revealed that primitive smooth muscle cells attached directly to the bulge initially at the mid-bulbous hair peg, the stage when the bulge is most prominent. K-laminin and type VII collagen are strongly expressed in the dermoepidermal junction of the bulge and between the matrix area of the bulb and the dermal papilla. Thus, the bulge of human hair follicle is not only an attachment site for arrector pili muscle, but also a pool of keratinocytes that are relatively undifferentiated. PMID- 7490482 TI - Nerve growth factor is increased in psoriatic skin. PMID- 7490484 TI - Mutations in the laminin 5 LAMB3 gene in generalized atrophic benign epidermolysis bullosa. PMID- 7490483 TI - Th2 cytokine profile in cutaneous T-cell lymphoma. PMID- 7490485 TI - Effects of Le Fort I osteotomy on human gingival and pulpal circulation. AB - The maxillary blood flow during the first 24 h following Le Fort I osteotomy was evaluated by laser Doppler flowmetry. Pulpal blood flow was recorded from two maxillary incisors and gingival blood flow was assessed from a site slightly apical to the interdental papilla of the maxillary central incisors of 12 patients receiving Le Fort I osteotomy, nine control patients receiving mandibular osteotomy, and 10 nonsurgical control subjects without orthodontic appliances. Measurements were made before surgery and at time intervals between 0 8, 8-16, and 16-24 h after surgery. The nonsurgical controls were tested at similar intervals. Gingival and pulpal blood-flow measurements did not change over time in the nonsurgical control group. Presurgical blood-flow values did not differ between the two surgical groups. Following surgery, mean gingival (but not pulpal) blood flow was significantly lower for patients treated with Le Fort I osteotomy than for patients treated with mandibular osteotomy. Follow-up examinations revealed that one patient receiving Le Fort I osteotomy experienced loss of gingiva and bone around both central incisors. This patient had one of the largest reductions in gingival blood flow. PMID- 7490486 TI - Mandibular resorption due to systemic sclerosis. Case report of surgical correction of a secondary open bite deformity. AB - Systemic sclerosis (SSc) is a connective-tissue disorder of unknown origin causing a multisystem disease. While erosions of the distal phalanges are commonly described, resorption of the mandible has been considered an unusual finding. However, systematic radiographic screening of different groups of patients suffering from SSc revealed a resorption incidence of 20-33% of the examined mandibles. Women especially seem to be affected, and the male/female ratio is 1/7. Bilateral condylysis due to SSc has been described in seven cases, or 13.7% of the reported cases. To the best of our knowledge, this is the fourth report of surgical correction of secondary dysgnathia due to systemic sclerosis and the first with a 2-year follow-up period. PMID- 7490488 TI - Bilateral mandibular subcondylar fractures contributing to airway obstruction. AB - Bilateral mandibular fracture contributing to airway obstruction is a well understood phenomenon. This is typically described as a rather immediate or acute process. After trauma, the production of a "flail mandible" with concomitant loss of support of tongue muscles is thought to result in obstruction of the upper airway. An unusual case in which the bilateral mandibular fractures contributed to an airway obstruction 10 days after the initial trauma is described. The cause is explained and methods of prevention and treatment are presented. PMID- 7490487 TI - Long-term effects of orthognathic surgery on the temporomandibular joint: comparison of rigid and nonrigid fixation methods. AB - Sixty-six patients were examined between 2 and 9.5 years after bilateral sagittal split mandibular ramus osteotomy. Thirty-two patients had nonrigid fixation consisting of superior border wires and intermaxillary fixation, while 34 patients had rigid plate fixation of the osteotomy sites with immediate function. There were no demonstrable long-term differences between the two groups with respect to mandibular vertical opening, crepitance, and temporomandibular joint pain. Masticatory muscle pain and temporomandibular joint clicking improved with rigid fixation and worsened with nonrigid fixation. PMID- 7490489 TI - Cavernous sinus-carotid fistula: a complication of maxillofacial injury. AB - A case of traumatic carotid-cavernous sinus fistula (CCSF) is presented. The patient sustained an undisplaced fracture of his right mandibular condyle and a zygoma fracture after a motor vehicle accident. Five months later, all the classic signs and symptoms of a CCSF had appeared except ophthalmoplegia. These symptoms were preceded by a tiring auditive homolateral sensation. A carotid angiography confirmed the presence of a large CCSF. Successful management was achieved intravascularly by balloon embolization. PMID- 7490494 TI - Congenital velopharyngeal incompetence in Kabuki make-up syndrome. AB - The Kabuki make-up syndrome is characterized by peculiar cranio-facial abnormalities, including cleft lip and palate. A case with velopharyngeal incompetence in a 7-year-old boy is reported. The significance of a possible association of this syndrome with congenital velopharyngeal incompetence is also discussed. PMID- 7490491 TI - Posterior ischemic optic neuropathy following bilateral radical neck dissection. AB - A patient is presented with posterior ischemic optic neuropathy (PION) after bilateral radical neck dissection, probably caused by hemodynamic hypotension, combined with other factors. A postmortem histologic study of the optic nerve was performed to analyze the pathogenic mechanism of blindness. To prevent this complication, one should favor a two-stage procedure, avoiding drug-induced hypotension, overtransfusion, and anemia. During the postoperative period, visual acuity should be monitored regularly, and proper positioning of the patient's head is necessary. PMID- 7490490 TI - The edentulous maxillary alveolar process in the region of the maxillary sinus--a study of physical dimension. AB - From 47 anatomic specimens, vertical sections were cut in the molar region, and computer-assisted measurements of both height and width of the alveolar ridge were made to assess the available bone volume for endosseous implant placement. The mean ridge heights ranged between 9.30 and 3.23 mm, the highest and lowest values being 13.8 and 0.8 mm, respectively. The ridge widths generally proved to be sufficient for placement of endosseous implants. Small knife-edge ridges are rarely found in the posterior maxilla. Reduction of the cancellous portion of the posterior maxilla seems to be influenced by the duration of edentulousness, as well as by osteoporotic changes, without being directly related to ridge configuration and bone volume. PMID- 7490492 TI - Mandibular reconstruction by secondary reimplantation of resected segments: a preliminary report. AB - This paper presents a new method of reconstructing the mandible after ablative surgery for cancer. This method uses autogenous grafts consisting of the respected part of the mandible, which is hollowed out, defatted in chloroform/methanol, sterilized with ethylene oxide, irradiated with beta radiation (dose 25 kGy), and stored for 7-10 months. At reimplantation, they were filled with compressed, autogenous, particulate cancellous bone. All grafts took, and four of six host-graft junctions healed. PMID- 7490493 TI - Salivary gland tumours in Zimbabwe: report of 282 cases. AB - Tumours of the salivary glands are relatively uncommon. In a review of 282 black patients seen at Harare Central Hospital, Zimbabwe, the relative incidence of various tumour types and the age and sex distribution were similar to those reported in other series. There were more tumours of the minor salivary glands than in reported Western series. There were more tumours of the minor salivary glands than in reported Western series. Pain and rapid growth were significant in distinguishing malignant from benign tumours. Malignant tumours were more common in elderly than in young patients. PMID- 7490495 TI - Angioleiomyoma of the upper lip: report of a case. AB - A rare case of vascular leiomyoma of the upper lip in a 51-year-old man is presented. The differential diagnosis, frequency, treatment, and prognosis are discussed. PMID- 7490499 TI - Bioengineering of cartilage in organ culture. PMID- 7490498 TI - Prevention of mandibular fractures by using constructional design principles. II. A tension strength test on beagle mandibles with two different types of segmental resections. AB - The tension strength of mandibular halves of beagle dogs, with two different types of segmental resections, was studied. It was found that the radius of the posterior resection corner is of utmost importance in reducing the risk of mandibular fractures. PMID- 7490496 TI - Delayed infantile cortical hyperostosis (Caffey's disease): case report. AB - Late recurrence or a delayed form of Caffey's disease is an exceedingly rare condition of unknown cause. A 3 1/2-year-old boy is presented with delayed Caffey's disease showing unilateral involvement of maxilla and zygoma. PMID- 7490497 TI - Prevention of mandibular fractures by using constructional design principles. I. Computer simulation of human mandibular strength after segmental resections. AB - A study was carried out on mechanical stress at the resection site in mandibles undergoing segmental resections. A computer simulation model was used to study particularly the effect of various radii of the posterior resection corners and the effect of elimination of muscle action. It was found that an enlarged radius of the resection corner in conjunction with coronoid process resection on the ipsilateral side offers the best condition to prevent fracture of the mandible. PMID- 7490500 TI - Late effects of radiation on the human immune system: an overview of immune response among the atomic-bomb survivors. AB - The studies of the late effects of atomic-bomb (A-bomb) radiation on the immune system were started about 20 years after the bombings in 1945. The most remarkable late effects of radiation are the functional and quantitative abnormalities of T and B cells in survivors exposed to high doses (> or = 1.0 Gy). Abnormalities of T-cell immunity include (1) a decreased proportion of CD3+ T cells in peripheral blood lymphocytes, particularly the proportion of CD4+ CD45RA+ naive T cells (study period 1987-91); (2) an increased frequency of CD4- and CD8- (double negative) alpha beta + T cells (1987-91); and (3) functional defects in T-cell responses to mitogens and alloantigens (1974-85). B-cell abnormalities include: (1) a significant increase in the proportion of B cells among peripheral lymphocytes (1987-91); (2) an increase in serum immunoglobulin A levels in females and immunoglobulin M and the incidence of rheumatoid factor in both sexes (1987-89); and (3) an increased level of anti-Epstein-Barr virus antibody titer (1987-90). In contrast, suggestive (0.05 < p < 0.1) or not significant (p > 0.1) dose effects were observed for the number and function of natural killer cells (1983-91), and benign monoclonal gammopathy (1979-87). In addition, studies initiated sooner after the bombing such as the incidence of autoimmune diseases (1958-87), systemic bacterial infections (1954-67), and granulocyte functions (1947-79) also show little dose-effects. Thus, A-bomb radiation induced the alteration of the balance/interaction between the T- and B cell subsets--specifically, a decrease in the T-cell population and an increase in the B-cell population in the periphery. PMID- 7490501 TI - Radiation chemistry of d(ApCpGpT). AB - The radiation chemistry of the DNA tetranucleoside triphosphate d(ApCpGpT) was investigated. Irradiations were carried out on aqueous solutions saturated with oxygen (with and without added Cu++), nitrogen or nitrous oxide. When oxygen was present, principal products were formed by hydroxylation at the 8-position of guanine and by degradation of thymine leaving a formamido remnant. Products were also formed containing both of the aforementioned lesions at adjacent deoxyguanosine and pyrimidine nucleosides. Other products resulted from rearrangement of the thymine ring generating two diastereoisomers of the 5-methyl 5-hydroxyhydantoin modification of d(ApCpGpT). Rearrangement of the cytosine ring occurred generating imidazolidine products and a hydantoin product. The product profiles are similar when either an N2O or N2 gaseous environment is maintained. However, in the latter case the dihydrothymine modifications of d(ApCpGpT) are markedly enhanced. Other products include an 8,5' cyclized product formed from the 2'-deoxyadenosine nucleoside and the 8-hydroxyguanine modification. 6-Hydroxy 5,6-dihydrothymine and 5,6-dihydroxy-5,6-dihydrothymine modifications of the thymidine nucleoside were also observed. A strand break product formed in oxygenated solution is also produced in nitrous oxide saturated solutions. Scission of the deoxyadenosine terminus was also observed. The effect of several of these lesions on d(ApCpGpT) as substrate for nuclease P1, bovine spleen phosphodiesterase and snake venom phosphodiesterase was studied. PMID- 7490503 TI - Inactivation of xanthine oxidase by oxidative radical attack. AB - Steady-state radiolysis, pulse radiolysis and epr studies, combined with enzyme activity measurements, were carried out on the mechanism by which radical attack, through one-electron oxidation, inactivates xanthine oxidase. Electron transfer to both the N3 and Br2- radical species was used to initiate oxidative damage on the enzyme. Inactivation was found to occur to a greater extent at low than at high pH and is associated with the initial formation of a tryptophanyl radical which converts by a known intramolecular pathway to a tyrosyl radical with a rate constant of 5 x 10(3) S-1. The tyrosyl radical in turn slowly loses around half of its absorbance at an intramolecular rate constant of 350S-1 and is consistent with the establishment of a radical equilibrium with cysteine residue(s). The sequence of reactions could be repeated several times on the same irradiated sample implying that restitution of the implied cysteinyl radical occurs leading to other damage in the protein. N3+Trp/N-->Trp/N-->Tyr/O<-->Cys/S-->?. Epr evidence implies that inactivation of the enzyme from the above sequence of reactions arises in part from alternations to Fe/S center I in the enzyme. PMID- 7490504 TI - Formation of 5-hydroxy-5,6-dihydrouracil and release of undamaged uracil as a result of poly(U) irradiation in N2O-saturated aqueous solution. AB - The radiation chemical yields of 5-hydroxy-5,6-dihydrouracil and uracil were measured on radiolysis of N2O-saturated solutions containing I mmol dm-3 poly(U) and were found to be 0.04 and 0.31 mumol J-1 respectively. From the 5-hydroxy-5,6 dihydrouracil yield in the presence of cysteamine and 2-mercaptoethanol, it is shown that only 20% of the OH-radicals, which attack poly(U) results in the formation of 5-hydroxy uracil adducts. It has been proposed that as yet unknown reducing radicals are formed in addition to known reducing radicals (the 5-OH adducts of uracil, the H-adducts of uracil, and sugar radicals) as a result of the reaction of OH-radicals with poly(U). PMID- 7490505 TI - Ionizing radiation-induced DNA strand breakage and rejoining in specific genomic regions as determined by an alkaline unwinding/Southern blotting method. AB - A recently developed, combined alkaline unwinding/Southern blotting assay was utilized to examine DNA damage and repair induced by ionizing radiation within specific large-scale genomic regions. Following treatment of MCF-7 breast tumour cells with 2-10-Gy gamma-rays, strand breakage and rejoining were measured in bulk DNA, in the centromeric alpha-satellite region of chromosome 17, and in the chromatin regions containing the unexpressed beta-globin gene and the expressed c myc oncogene, which is known to be important for growth in the MCF-7 cell line. Damage in both the c-myc and beta-globin regions was markedly greater than in either alpha-satellite or bulk DNA. However, the kinetics of strand break repair were approximately the same in c-myc as in alpha-satellite or bulk DNA. Surprisingly, the radiomimetic antibiotic bleomycin, which also induces free radical-mediated strand breakage, showed considerably less heterogeneity of DNA damage among the genomic regions examined than did radiation. The results suggest that actively transcribed genes, as well as at least some inactive genes, are surrounded by large-scale domains of radiosensitive chromatin. With no apparent enhancement of rejoining, the increased incidence of strand breaks in these regions persists until rejoining is essentially complete. Changes in the integrity of specific chromatin regions may be an important aspect of DNA damage induced cell death. PMID- 7490502 TI - Inhibition of OH radical-induced strand break formation of poly(U) by Ru(bpy)32+ or Ru(phen)32+ attached to the polynucleotide. AB - Reactions of OH radicals with poly(U) (polyuridylic acid) in the presence of Ru(bpy)32+ or Ru(phen)32+ in aqueous solutions were studied. OH radicals were produced by pulse radiolysis and their reactions with ruthenium complexes were measured spectrophotometrically under conditions were the complexes are attached to the polynucleotide. The OH radical adds to either the uracil moiety or the ruthenium complexes. The ratio of the radicals produced depends only on the ratio of their rate constants and the concentrations of poly(U) and ruthenium complexes. Similar results were obtained with uridine-5'-monosphosphate, where the ruthenium complexes are not attached to the nucleotide. Surprisingly, the yield of single-strand break formation from the OH adducts of uracil in poly(U) is much smaller than that expected on the basis of the yield measured in the absence of ruthenium complexes. Possible reasons for this behaviour are discussed. PMID- 7490506 TI - Induction of DNA single-strand breaks in human lymphocytes by low doses of gamma rays. AB - We hypothesized that very low doses of gamma-rays have a genotoxic effect as measured by DNA single-strand breaks (ssbs). To test our hypothesis we used a newly developed alkaline microgel electrophoretic technique to quantitate DNA ssbs in freshly isolated human lymphocytes. An enhanced sensitivity to detect low levels of DNA damage was made possible by this method due to an additional step of DNA precipitation by ethanol in agarose microgels. We exposed lymphocytes from three subjects for 1 h at 4 degrees C to 740 MBq of technetium 99-m at a distance of 20 cm (0.25 mGy), 10 cm (1.02 mGy), and 5 cm (4.08 mGy) from the source vial. Doses of 1.02 and 4.08 mGy induced a significant dose-dependent increase in DNA ssbs in human lymphocytes from three subjects, each examined on three different days. PMID- 7490508 TI - Vitamin E prevents UVB-induced cell blebbing and cell death in A431 epidermoid cells. AB - Cultured A431 epidermoid cells exposed to UVB (120-2400 J/m2) develop numerous blebs on their surface, detach from the plastic dish, and undergo injury and death. Numerous detached cells display fragmented nuclei, typical of apoptotic cells. Since bleb formation also occurs after oxidative stress it was assumed that the morphological variations observed are the consequence of free radical mediated insult. In order to test this hypothesis, the antioxidant alpha tocopherol (vitamin E) was added to cell cultures at different times, before or after irradiation. The results indicate that vitamin E inhibits UVB-induced surface blebbing as well as cell detachment from the substrate. Moreover, vitamin E is most effective in stimulating cell recovery when it is added after the end of UVB irradiation. Finally, vitamin E treatment also seems to reduce the fraction of cells undergoing death (probably those which will undergo apoptosis) after exposure to UVB radiation. PMID- 7490507 TI - Radioprotection of DNA by polyamines. AB - Putrescine, spermidine and spermine are natural polyamines bearing at neutral pH the net electrical charges +2, +3 and +4 respectively. We report here the radioprotective effect of these polyamines on the radiolysis of pBR322 plasmid DNA. We observe a very efficient protection against fast neutron-induced single and double-strand breakage in the presence of spermine and spermidine, and a significantly less efficient protection in the presence of putrescine. An ionic strength dependence is observed for spermidine and spermine, but not for putrescine. Circular dichroism measurements show spermidine- and spermine-induced structural modifications of DNA, i.e. the formation of tightly packaged condensates in the concentration range corresponding to radioprotection. No structural change is observed for concentrations of putrescine affording radioprotection. We explain the radioprotection by: (1) the scavenging of OH radicals in the bulk, essentially observed in the case of putrescine; (2) a local scavenging at the sites of binding of polyamines; and (3) the reduced accessibility of the attack sites in the condensed structures induced by spermine or spermidine. PMID- 7490509 TI - Autoepitopes in lupus. PMID- 7490510 TI - Cytokine gene regulation by viral gene products. PMID- 7490511 TI - Catabolism of rabbit prothrombin in rabbits: uptake of prothrombin by the aorta wall before and after a de-endothelializing injury in vivo. AB - After an injury to the vascular endothelium, certain blood proteins collect rapidly at the site of damage to prevent blood loss and maintain blood flow. The uptake of fibrinogen, plasminogen, and antithrombin--but not prothrombin--have been measured previously at the rabbit aorta wall after injury in vivo. This report describes the clearance of rabbit iodine 131-labeled prothrombin from the rabbit circulation to measure the distribution and fractional catabolic rate and compares the behavior of 131I-labeled prothrombin with either iodine 125-labeled fibrinogen or 125I-labeled antithrombin at the balloon catheter-injured aorta wall. When injected into young rabbits, 131I-labeled prothrombin was cleared from the intravascular space to yield a plasma curve that was best described by three exponentials. Fractional plasma and whole body catabolic rates were 2.0 day-1 and 0.41 day-1, respectively, equivalent to a catabolic half-life of 1.7 days. Fractional distribution of prothrombin amounted to 0.21, 0.24, and 0.55 within the intravascular, vascular endothelial, and extravascular compartments, respectively. Samples of 131I-labeled prothrombin and either 125I-labeled fibrinogen or 125I-labeled antithrombin were injected into anesthetized rabbits before balloon de-endothelialization of the thoracic aorta. The uptake of each radiolabeled protein by the aorta intima-media was measured at various times (5 to 60 minutes) after injury. Whereas uptake of plasma fibrinogen by the balloon injured intima-media was maximal (20 pmol/cm2) in less than 5 minutes after injury, maximum uptake of prothrombin (5 to 6 pmol/cm2) took approximately 15 minutes. Uptake of prothrombin was initially faster than that of antithrombin, although approximately equimolar amounts of prothrombin and antithrombin were bound by the intimamedia by 60 minutes. The results are discussed in relation to thrombin production and the demand for antithrombin by the damaged aorta wall in vivo. PMID- 7490512 TI - Tryptophan metabolism in chronic inflammatory lung disease. AB - Induction of indoleamine 2,3-dioxygenase (IDO), an enzyme expressed by mononuclear phagocytes and some fibroblast cell lines in response to interferon gamma, leads to enhanced degradation of tryptophan to kynurenine. Because inflammatory lung diseases are generally associated with activation of pulmonary macrophages, we investigated tryptophan metabolism in patients with interstitial lung disease by measuring circulating levels of tryptophan and kynurenine in peripheral blood and by measuring the IDO activity of bronchoalveolar cells. IDO activities were increased for bronchoalveolar lavage (BAL) cells obtained from patients with interstitial lung disease (115.4 +/- 30.4, n = 37) when compared with BAL cells from normal subjects (15.2 +/- 7.4, n = 14; p < 0.05), and messenger RNA for IDO was present in BAL cells from patients with interstitial disease but was not present in BAL cells from normal volunteer subjects. Patients with inflammatory lung disease also had decreased tryptophan and increased kynurenine concentrations in serum. The ratio of serum tryptophan levels to serum kynurenine levels was significantly depressed for patients with idiopathic pulmonary fibrosis (18.4 +/- 1.7, n = 29; p < 0.0001), patients with fibrosing alveolitis associated with collagen vascular disease (13.1 +/- 1.6, n = 18; p < 0.0001), or patients with sarcoidosis (21.0 +/- 1.1, n = 50; p < 0.0001), as compared with the ratio for normal subjects (31.8 +/- 2.3, n = 18). Patients with fibrotic disease had the highest levels of BAL cell IDO activity, and patients with collagen vascular disease associated fibrosing alveolitis had the most depressed levels of serum tryptophan and the greatest elevations in serum kynurenine. Measurement of tryptophan and kynurenine concentrations in serum may provide a useful measure of disease activity in chronic inflammatory parenchymal lung diseases such as sarcoidosis and idiopathic pulmonary fibrosis. PMID- 7490514 TI - Protective effects of reconstituted high-density lipoprotein in rabbit gram negative bacteremia models. AB - Reconstituted high-density lipoproteins (rHDLs) have the ability to bind bacterial lipopolysaccharide and to reduce its endotoxin activity in vitro and in vivo. The aim of the present studies was to investigate the therapeutic potential of rHDL in bacteremia models. Gram-negative sepsis was induced in anesthetized rabbits by intravenous infusion of Escherichia coli organisms (4 x 10(9) CFU/kg infused over 2 hours) and treated with appropriate antibiotics. rHDL or placebo was infused either before (prophylaxis) or 1 hour after (therapy) the beginning of the bacterial challenge. In the control groups, the bacterial challenge resulted in transient bacteremia, high plasma levels of lipopolysaccharide, secretion of TNF, and symptoms of sepsis, including hypotension and acidosis. rHDL had no influence on blood bacterial counts; however, plasma lipopolysaccharide levels were significantly reduced. Peak plasma TNF concentrations were reduced after prophylactic but not after therapeutic rHDL administration. Both prophylactic and therapeutic rHDL improved clinical outcome: acidosis was significantly attenuated and blood pressure tended to be higher in the rHDL groups. No effects of rHDL were seen in a similar model of gram-positive sepsis induced by the infusion of Staphylococcus aureus. PMID- 7490516 TI - Biphasic granulocytopenia after administration of the first dose of OKT3. AB - The effects of the administration of OKT3, a second immunoglobulin G2a (IgG2a) anti-CD3 monoclonal antibody (mAb), and its isotype switch variant IgA on granulocyte kinetics were compared for 5 hours after the first administration of the mAb. In addition, in vivo and in vitro studies were performed on alterations in expression of CD11b and CD62L induced by these mAbs. Within 15 minutes after administration OKT3 and IgG2a anti-CD3 mAbs induced a significant decrease in circulating granulocytes, whereas IgA anti-CD3 mAb did not. Apparently the initial decrease in circulating granulocytes depends on the heavy chain of the administered anti-CD3 mAb, resulting in immunocytoadherence and sequestration in the lungs. Increased adherence to pulmonary endothelium by altered expression of CD11b and CD62L plays a minor role in this first granulocytopenia, because each mAb exerted the same effects on these adhesion molecules in vitro. The second decrease in granulocyte counts occurred 60 minutes after administration of each mAb and correlated with a significant increase in expression of CD11b and CD62L in vivo and with upregulation of CD11b and down-regulation of CD62L in vitro. These alterations could be related to the presence of tumor necrosis factor-alpha both in vivo and in vitro. Thus granulocyte kinetics from 30 minutes after administration of each anti-CD3 mAb resemble neutrophil kinetics induced by TNF alpha. PMID- 7490515 TI - Role of endothelin-converting enzyme in the systemic hemodynamics and regional circulatory effects of proendothelin-1 (1-38) and diaspirin cross-linked hemoglobin in rats. AB - Diaspirin cross-linked hemoglobin (DCLHb) is a promising hemoglobin-based, oxygen carrying resuscitative solution. DCLHb (400 mg/kg, iv) produces significant cardiovascular effects, along with an increase in plasma endothelin-1 (ET-1) level, when administered to conscious or anesthetized rats. Present studies were performed to determine whether the cardiovascular effects of DCLHb are due to an increase in the conversion of proendothelin-1 (1-38) (proET-1) to ET-1 by endothelin-converting enzyme (ECE). The regional circulatory and systemic hemodynamic effects of proET-1 (20 micrograms/kg, iv) and DCLHb (400 mg/kg, iv) were determined by using a radioactive microsphere technique in control rats and rats pretreated with phosphoramidon (ECE inhibitor). Administration of proET-1 produced an immediate increase in mean arterial pressure (MAP)(52%) and total peripheral resistance (TPR) (55%); stroke volume (SV) and cardiac output were not affected in the initial phase but were decreased subsequently. Heart rate (HR) was not affected after administration of proET-1. A significant increase in blood flow to the heart (39%), brain (46%), kidneys (74%), portal system (40%), and gastrointestinal tract (GIT) (42%) was also observed after administration of proET-1. Vascular resistance was found to be significantly increased in the mesentery and pancreas (168%) and in the musculoskeletal system (147%) and decreased in the kidneys (-11%) after administration of proET-1. Phosphoramidon (4 mg/kg, iv) pretreatment attenuated the increase in MAP and TPR induced by proET-1. Phosphoramidon pretreatment significantly attenuated the proET-1-induced increase in blood flow to the heart, brain, kidneys, portal system, and GIT. The increase in vascular resistance induced by proET-1 in the mesentery and pancreas and in the musculoskeletal system was also attenuated by phosphoramidon. DCLHb increased MAP (63%) and TPR (54%) without affecting HR. DCLHb increased blood flow to the heart (95%), GIT (45%), portal system (43%), and skin (79%) and increased vascular resistance in the musculoskeletal system (58%). In phosphoramidon-treated rats, DCLHb increased MAP (99%), HR (25%), cardiac output (37%), and TPR (60%). DCLHb increased blood flow to the heart (104%), brain (66%), kidneys (49%), GIT (59%), portal system (63%), and skin (100%) when administered to phosphoramidon-treated rats. Phosphoramidon did not attenuate any of the DCLHb-induced cardiovascular effects. It is concluded that proET-1 increases blood flow to various organs and that phosphoramidon, an ECE inhibitor, could block the proET-1-induced increases in regional blood flow.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 7490513 TI - An aldose reductase inhibitor, TAT, reduces ADP-induced platelet hyperaggregation in streptozotocin-induced diabetic rats with neuropathy. AB - To investigate the relationship between metabolic and vascular factors, especially polyol pathway and platelet aggregation, in the pathogenesis of diabetic neuropathy, the effects of a novel potent aldose reductase inhibitor, TAT ((5-(3-thienyl) tetrazol-1-yl) acetic acid monohydrate) on adenosine diphosphate-induced platelet aggregation, polyol contents in platelets, motor nerve conduction velocity (MNCV), and sciatic nerve blood flow (SNBF) were examined in streptozotocin-induced diabetic rats. Diabetic rats demonstrated hyperaggregation in response to adenosine diphosphate, accompanied by sorbitol and fructose accumulation and myoinositol depletion in platelets. Treatment with TAT improved these abnormalities in diabetic rats. A delayed MNCV and a reduced SNBF in diabetic rats were normalized by the administration of TAT. These observations suggest that increased polyol pathway activity plays an important role in platelet aggregation in the development of diabetic neuropathy and that aldose reductase inhibitor is useful for the treatment of diabetic neuropathy from the viewpoint not only of metabolic factors but also of vascular factors. PMID- 7490517 TI - Evidence for E-selectin complement regulatory domain mRNA splice variants in the rat. AB - The adhesion protein E-selectin is one mediator of endothelial cell-leukocyte interaction during acute inflammation. To investigate the molecular regulation of E-selectin function, we have examined the expression of E-selectin mRNA in target rat tissues after administration of lipopolysaccharide, a potent inducer of acute inflammation. In the course of these studies we isolated two unique rat E selectin cDNA fragments. Both cDNA fragments show extensive nucleotide sequence homology to previously isolated mouse and human E-selectin cDNAs. However, they differ for the presence of sequences that encode complement regulatory domain-5 (CR5). Previous studies have shown that different animal species express E selectin mRNAs that encode different numbers of CR domains. The isolation of these two rat E-selectin cDNA fragments, which differ only for the presence of CR5, represents the first direct evidence for the existence of E-selectin CR variant mRNAs in the same species. Moreover, the sequence of the CR5(-) cDNA is consistent with its origin from an mRNA splice variant of a CR5(+) mRNA. We have demonstrated the presence of the two predicted mRNA species in rat heart tissue and have investigated their expression in response to lipopolysaccharide. Although both mRNA variants were greatly induced by lipopolysaccharide, the CR5( ) form was more abundant in both treated and control tissues. This difference in mRNA abundance may indicate different levels of CR5 variant proteins that perform functionally distinct tasks in E-selectin dependent inflammatory processes. PMID- 7490518 TI - Neutrophil activation in response to immune complex-bearing endothelial cells depends on the functional cooperation of Fc gamma RII (CD32) and Fc gamma RIII (CD16). AB - Leukoclastic vasculitis is thought to be initiated by deposition of immune complexes (ICs) in the vascular wall. To study the neutrophil response in a related in vitro model, we primed human umbilical vein endothelial cell (HUVEC) monolayers with antibodies against human fibronectin. The resulting respiratory burst to the immobilized ICs depended on the antibody concentration used to prime the monolayers and included a marked release of primary and secondary granule constituents. On IC-bearing HUVEC monolayers, but not on ICs directly bound to tissue culture dishes, blocking monoclonal antibodies (mAbs) to crystallizable fragment-gamma receptor II (Fc gamma RII) and Fc gamma RIII markedly inhibited the respiratory burst and the release of elastase. However, on both surfaces the neutrophil response was strongly inhibited by mAbs against CD18. Regardless of whether we used neutrophils from a patient with severe paroxysmal nocturnal hemoglobinuria (PNH) lacking the Fc gamma RIII, or whether the Fc gamma RII mediated signal transduction was blocked by pertussis toxin, the respiratory burst to the IC-bearing HUVECs was essentially unchanged. With PNH neutrophils, the respiratory burst was predominantly blocked by an anti-Fc gamma RII mAb. In contrast, the response of pertussis toxin treated neutrophils was strongly inhibited by a mAb against Fc gamma RIII. Together these data indicate that the answer of neutrophils to ICs immobilized at the endothelial barrier depends on the cooperative function of both low-affinity Fc gamma Rs. PMID- 7490519 TI - Hepatic mitochondrial oxidative metabolism and lipid peroxidation in iron-loaded rats fed ethanol. AB - The aims of this study were to determine whether chronic ethanol consumption potentiates mitochondrial lipid peroxidation or impairment of mitochondrial oxidative metabolism in rats with chronic iron overload. Experimental iron overload was induced by feeding rats a chow diet supplemented with 2.5% carbonyl iron. After 8 to 12 weeks, half of the iron-loaded and control animals were changed to a liquid diet containing ethanol for 4 to 5 weeks. The remaining animals were fed an isocaloric amount of diet containing dextrin-maltose instead of ethanol for 4 to 5 weeks. Iron-supplemented animals had a 20-fold increase in hepatic iron concentration as compared with controls. Iron and ethanol independently increased plasma alanine aminotransferase (ALT) levels (p < 0.05) while the combination resulted in an additive increase in ALT levels (p < 0.01). Although iron overload increased the levels of mitochondrial conjugated dienes and significantly reduced the mitochondrial respiratory control ratio, ethanol administration did not affect these parameters in animals with or without iron overload. Livers from iron-loaded rats that received ethanol showed mild to moderate steatosis with scattered necroinflammatory foci. There was no significant increase in necroinflammatory foci in the livers of the iron plus ethanol group as compared with the iron group. In conclusion, we have demonstrated an additive increase in hepatocellular injury when ethanol is fed to iron-loaded rats, as evidenced by an increase in plasma ALT level. However, there were no additive or synergistic effects of iron and ethanol on either mitochondrial lipid peroxidation or mitochondrial oxidative metabolism. PMID- 7490521 TI - The neuroendocrine bag cells of Aplysia: a model system for neural control of hormone secretion. PMID- 7490520 TI - Development of a whole platelet ELISA to detect circulating activated platelets. AB - P-selectin is a granule membrane protein that is expressed on the surface of activated endothelial cells and platelets. Flow cytometry has been used as a means of detecting activated platelets in the circulation by using antibodies to P-selectin and other surface markers. In the study reported here, we developed a whole platelet ELISA for measuring P-selectin on platelets in platelet-rich plasma. Platelet-rich plasma samples for analysis were isolated from fresh blood by centrifugation, fixed with 1.0% paraformaldehyde, and used within 3 hours or after storage at -70 degrees C for up to 10 months. Paraformaldehyde-fixed, phorbol myristate acetate-activated or thrombin receptor peptide-activated platelets were used to construct a standard calibration curve. These platelets were stable after 10 months of storage at -70 degrees C. Interassay variability showed a high degree of correlation, with r = 0.98 +/- 0.03 (n = 12). The accuracy and specificity of the ELISA was verified by using fluorescence activated flow cytometric analysis and is as sensitive (< or = 0.5%) as flow cytometry for detecting P-selectin expression on platelets. To assess the ability of the platelet ELISA to detect platelet activation in the systemic circulation, we examined 24 patients with unstable angina and 12 age-matched control subjects. Patients with unstable angina demonstrated significantly higher levels of circulating activated platelets than did age-matched control subjects. Although storage-dependent differences in absolute platelet activation levels were found, platelet ELISA results of samples evaluated within either 3 hours or after 10 months of storage were comparable to results obtained by fluorescence-activated flow cytometric analysis.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7490522 TI - Interaction between activin A and cAMP in the induction of FSH receptor in cultured rat granulosa cells. AB - The acquisition of FSH receptor during preantral folliculogenesis is believed to be a key step in the subsequent development of follicles. We examined the interaction between activin and cAMP in FSH receptor induction in rat granulosa cells by measuring 125I-FSH binding and FSH receptor mRNA. In the 125I-FSH binding study, 0.2 mM 8-Br-cAMP and 1 microM forskolin were maximally effective in FSH receptor induction (169 and 220% respectively of control), while higher concentrations gave attenuated responses. It appears that cAMP has ambivalent effects on FSH receptor induction depending on the concentration and length of exposure. Activin alone dramatically increased the number of FSH receptors (314% of control). Moreover, synergistic effects of activin and 8-Br-cAMP or forskolin were observed on FSH receptor induction: a combination of activin (80 ng/ml) and low doses of 8-Br-cAMP (0.2 mM) or forskolin (1 microM) was most effective (160 or 140% of that induced by activin alone) and receptor levels reached a maximum at 24 h. These levels than markedly decreased after 72 h of incubation. Northern blot analysis revealed that the combination of activin (80 ng/ml) and 8-Br-cAMP (0.2 mM) or forskolin (1 microM) increased FSH receptor mRNA to about 140% of that induced by activin alone. These results indicate that activin and cAMP induced FSH receptor synergistically. However, activin did not enhance the production of cAMP induced by forskolin. In addition, a protein kinase A inhibitor (H89) (2 microM), which inhibited the effects of forskolin, had no effect on the action of activin.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7490523 TI - Cytokine receptor signalling through two novel families of transducer molecules: Janus kinases, and signal transducers and activators of transcription. PMID- 7490524 TI - Changes in testicular function following specific deprivation of LH in the adult male rabbit. AB - Sexually mature male rabbits actively immunized against highly purified ovine LH (oLH) were used as a model system to study the effects of endogenous LH deprivation (and therefore testosterone) on spermatogenesis as well as pituitary FSH secretion. Immunization against oLH generated antibody titres capable of cross-reacting and neutralizing rabbit LH and this resulted in a significant reduction (P < 0.01) in serum testosterone levels by 2-4 weeks of immunization. A significant increase in circulating FSH concentration (from a basal level of approximately 1 ng to 60-100 ng/ml; P < 0.01) was observed within 4-6 weeks of immunization, perhaps a consequence of the negative feedback effect of the lack of testosterone. The effect of LH deprivation on spermatogenesis assessed by DNA flow cytometry and histological analyses of testicular biopsy tissue revealed that lack of testosterone primarily results in a rapid reduction and complete absence of round (1C) and elongated (HC) spermatids. The immediate effect of LH/testosterone deprivation thus appears to be at the step of meiotic transformation of primary spermatocytes (4C) to 1C. A significant reduction (> 80%; P < 0.01) in the 4C population and a relative accumulation (> 90%; P < 0.01) in spermatogonia (2C) was also observed, suggesting a need for testosterone during the transformation of 2C to 4C. In all but one of the rabbits, both qualitative and quantitative recovery in spermatogenesis occurred during the recovery phase, even at a time when only a marginal increase in serum testosterone (compared with the preimmunization) levels was observed as a result of a rapid decline in the cross-reactive antibody titres. These results clearly show that LH/testosterone deprivation in addition to primarily affecting the meiotic step also regulates the conversion of 2C to 4C during spermatogenesis. PMID- 7490525 TI - A 67 kDa protein mediates pituitary adenylate cyclase-activating polypeptide and vasoactive intestinal peptide-stimulated insulin secretion in a hamster clonal beta-cell line. AB - Insulin secretion is regulated by neural and neurohormonal factors. The report of nerves releasing pituitary adenylate cyclase-activating polypeptide (PACAP)--a 38 amino acid peptide--in the endocrine pancreas, suggests it may be important in modulating insulin release. We therefore carried out receptor-binding studies on membranes prepared from the glucose-responsive clonal beta-cell line HIT-T15, and also examined the effects of PACAP38, PACAP27--a C-terminal truncated form of the peptide--and vasoactive intestinal peptide (VIP) on insulin and islet amyloid polypeptide (IAPP) release. We showed by chemical cross-linking that PACAP and VIP stimulate secretion from the clonal cells by binding to a receptor with a molecular weight of 67 kDa (n = 4). Binding was saturable when membranes were incubated with 125I-PACAP27 (Kd 1.2 +/- 0.2 nM; Bmax 415.7 +/- 35.3 fmol/mg; n = 4) or 125I-VIP (Kd 1.3 +/- 0.3 nM; Bmax 354.8 +/- 42.8 fmol/mg; n = 4). We also demonstrated an increase in glucose-stimulated insulin (PACAP27, 366.6 +/- 25.8%; PACAP38, 389.9 +/- 13.4%; VIP, 342.6 +/- 16.1% of control; all at 1 microM, P < 0.01 vs control) and IAPP release (PACAP27, 236.9 +/- 26.2%; PACAP38, 226.5 +/- 10.9%; VIP, 242.9 +/- 15.8% of control; all at 1 microM, P < 0.01 vs control). Incubation of the cells with these peptides, for a duration of 12 h, in the presence of 5.5 mM glucose, did not alter the expression of insulin or IAPP. These findings suggest that PACAP and VIP stimulate secretion of insulin and IAPP by binding to a 67 kDa protein on clonal beta-cells and do not alter the transcription of insulin and IAPP under the conditions tested. PMID- 7490526 TI - Induced luteal regression in the primate: evidence for apoptosis and changes in c myc protein. AB - There is increasing molecular evidence that apoptosis is involved in the process of structural luteal regression in non-primate species. Apoptosis is dependent upon the activation of certain proto-oncogenes and c-myc protein has an important regulatory role in this process in some cell types. The aim of the present study was to determine the occurrence and localisation of c-myc protein within the primate corpus luteum, determine changes during induction of luteal regression and examine the corpora lutea for morphological evidence of apoptosis. Ovaries were studied from marmoset monkeys in the late follicular, and in the early, mid and late luteal phases. Luteal regression was induced either by treatment with prostaglandin F2 alpha analogue or GnRH antagonist administered during the mid luteal phase and ovaries obtained 24 and 48 h later. Immunocytochemistry was performed using a monoclonal antibody to the c-myc protein. In pre-ovulatory follicles positive staining was found in the nucleus of a few granulosal cells and in the cytoplasm of thecal cells. c-myc was present in all corpora lutea where it was localised predominantly in the cytoplasm. In early corpora lutea, scattered cells with intense staining were observed in the presence of a majority of moderately or weakly stained cells. In the mid and late luteal phases, corpora lutea were uniformly moderately stained for c-myc. Following induction of luteal regression, regression, nuclear degeneration with condensation and fragmentation indicative of apoptosis was observed. In other luteal cells, increased membranes suggested necrosis.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7490527 TI - Levels of pro-opiomelanocortin and prolactin mRNA in the fetal sheep pituitary following hypoxaemia and glucocorticoid treatment in late gestation. AB - It is well established that corticotrophin-releasing hormone and vasopressin can induce both synthesis and release of ACTH from the ovine pituitary gland, and that glucocorticoids can inhibit these responses. Changes in the abundance, localization and distribution of pro-opiomelanocortin (POMC) mRNA and prolactin (PRL) mRNA in the ovine fetal pituitary were examined by in situ hybridization following hypoxaemia applied in the presence or absence of concomitant cortisol in late gestation (day 135). Fetuses were distributed amongst four groups; saline infused/normoxaemic, cortisol-infused/normoxaemic (0.3 mg/h), saline infused/hypoxaemic and cortisol-infused/hypoxaemic. Hypoxaemia (6 h) was induced by reducing the maternal PaO2, resulting in a 6-8 mmHg decrease in fetal arterial PO2. Fetal infusions were commenced 5 h prior to and maintained throughout the treatment period. Hypoxaemia, which elevated fetal plasma ACTH and cortisol, caused a significant (P < 0.05) increase in POMC mRNA in the pars distalis (PD), but was without effect on POMC mRNA in the pars intermedia (PI). Cortisol infusion attenuated the hypoxaemia-induced increase in POMC mRNA in the PD, but was without effect on non-stimulated steady-state POMC mRNA levels in either the PD or PI. PRL mRNA was only present in the PD and significantly (P < 0.05) increased after cortisol infusion and hypoxaemia. In conclusion (i) POMC and PRL mRNA in the PD are increased following moderate hypoxaemia, (ii) cortisol attenuates changes in POMC mRNA but not PRL mRNA in the PD following hypoxaemia and (iii) cortisol increases PRL mRNA levels in the PD.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7490529 TI - Charge heterogeneity of the AT1 angiotensin II receptor subtype in the rat lung. AB - In order to obtain more information on the molecular structure of the angiotensin II (Ang II) binding sites from whole rat lung membranes these were characterized by isoelectric focusing (IEF) and SDS-PAGE. Whereas a single population of Ang II receptor sites was identified (Kd = 2.2 +/- 0.3 nmol/l; Bmax = 203.9 +/- 15.8 fmol/mg protein) by Scatchard analysis, using IEF three Ang II binding isoforms were observed; a major band which migrated to isoelectric point (pI) 6.7, and two minor bands with pI values of 6.5 and 6.3. Specific binding of 125I-Ang II to rat lung membrane preparations was sensitive to Losartan, a non-peptide AT1 receptor subtype antagonist, but was unaffected by the AT2 receptor subtype antagonist CGP42112A. Immunoblotting analyses on SDS gels, using a monoclonal antibody specific to the AT1 receptor, showed two immunoreactive protein species of 45 and 48 kDa. Enzymic deglycosylation using recombinant N-glycanase did not alter the molecular weight patterns of the AT1 receptor subtype. The results of the present study demonstrated that the Ang II receptor population in the whole rat lung consists solely of the AT1 receptor subtype and that the AT2 receptor subtype is absent. In addition, the data showed the existence of charge heterogeneity of the AT1 receptor subtype, and suggest that glycosylation probably does not contribute to its charge heterogeneity. PMID- 7490528 TI - Additive effects of growth hormone and testosterone on lipolysis in adipocytes of hypophysectomized rats. AB - The effects of growth hormone (GH) and testosterone, alone or in combination, on the regulation of lipolysis in isolated adipocytes from hypophysectomized rats were investigated. Male Sprague-Dawley rats were hypophysectomized at 50 days of age. One week after operation, hormonal replacement therapy with L-thyroxine and hydrocortisone acetate was given to hypophysectomized rats. Groups of rats were treated with GH (1.33 mg/kg, daily), testosterone (10 mg/kg, once) alone or in combination. After one week of hormonal treatment, adipocytes were isolated from the pooled epididymal and perirenal fat pads and glycerol release after isoproterenol stimulation and 125I-cyanopindolol binding was measured. Hypophysectomy caused a marked decrease in basal and isoproterenol-stimulated lipolysis. There was no effect of testosterone treatment alone on lipolysis, but GH treatment resulted in an increase in isoproterenol-induced lipolysis but not to the levels observed in cells from control rats. Testosterone and GH in combination restored the lipolytic response to isoproterenol. Also 125I cyanopindolol binding was decreased after hypophysectomy. Testosterone treatment alone and GH treatment alone increased the binding, while in combination the treatment had an additive effect. Affinity was not changed, but the effects seemed to be on receptor number, as determined by Scatchard analysis. Forskolin stimulated cAMP accumulation in adipocytes was markedly reduced after hypophysectomy. Testosterone treatment alone had no effect. GH treatment alone increased forskolin-stimulated cAMP accumulation, although the level was lower than that found in control rats. The combined treatment resulted in a further increase to levels observed in adipocytes from control rats.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7490530 TI - Evidence for axonal sprouting in the anterior pituitary following adrenalectomy in the rat. AB - Our recent studies have demonstrated the presence of peptidergic nerve fibers in the anterior pituitary of the rat. They were found to increase in number following adrenalectomy and the present study was aimed at investigating whether axonal sprouting could account for this increase. Antibody against the neuronal growth-associated protein GAP-43 was used as the probe. Four days following adrenalectomy GAP-43-like immunoreactivity was found to increase dramatically, mostly as varicosities surrounding the gland cells. The results suggest an active axonal sprouting following this hormone manipulation and strongly support our hypothesis of neural-humoral dual regulation of the mammalian anterior pituitary. PMID- 7490532 TI - Functional luteolysis in response to hydrogen peroxide in human luteal cells. AB - To evaluate the effect of reactive oxygen species in human corpus luteum function, we investigated whether hydrogen peroxide (H2O2) affects the in vitro luteal cell production of steroids. H2O2 treatment (1.0-100 microM) of mid and late luteal cell cultures elicited a dose-dependent decrease in basal progesterone production. However, treatment of mid luteal cells with a low concentration of H2O2 (0.01 microM) significantly stimulated progesterone secretion (P < 0.05). In addition, H2O2 (100 microM) markedly inhibited human chorionic gonadotropin (hCG)-stimulated progesterone and estradiol secretion. cAMP production was enhanced (2.4-fold, P < 0.05) by hCG treatment of luteal cells. The addition of H2O2 (0.1-100 microM) to hCG-stimulated luteal cell cultures elicited a decrease in cAMP concentration (P < 0.05) and in the specific binding of radiolabeled hCG by luteal cells. Progesterone and estradiol production stimulated by dibutyryl cAMP were significantly inhibited by H2O2 (P < 0.05). These findings suggest that H2O2 interferes with basal steroid production and, in hCG-stimulated conditions, it may inactivate the gonadotropin-receptor complex. The anti-steroidogenic action of H2O2 therefore raises the possibility of a modulatory role of H2O2 in human luteal steroidogenesis. PMID- 7490531 TI - Tumour necrosis factor alpha, interferon gamma and dexamethasone regulate IGF-I maintained collagen production in cultured human fibroblasts. AB - We examined the effects of tumour necrosis factor alpha (TNF alpha) and interferon gamma (IFN gamma) on the production of collagen by human infant foreskin fibroblasts. Collagen synthesis was maintained in the presence of IGF-I so that cytokine effects could be examined in the absence of serum. TNF alpha inhibited IGF-I-maintained collagen production in a dose-dependent manner. Maximal suppression of 50% was attained at a concentration of 7.5 ng/ml. IFN gamma also suppressed collagen accumulation in IGF-I-maintained cells, with a maximal inhibition to 55% at 375 U/ml. The rate of collagen formation relative to total protein production for secreted proteins was calculated. This value decreased from 10.3% for IGF-I-cultured cells to 6.2% and 8.4% with the inclusion of TNF alpha and IFN gamma respectively, indicating that inhibition was selective for collagen. TNF alpha (5 ng/ml) and IFN gamma (250 U/ml) together suppressed IGF-I-maintained collagen production to approximately 35%, with a decrease from 10.3% to 2.9% in collagen production relative to total protein. The inclusion of a serum-free period prior to the addition of TNF alpha to the cultures resulted in a further inhibition to 15% of control. This increase in inhibition was not seen if dexamethasone was present in the serum-free period prior to cytokine addition. These data showed that IGF-I-maintained collagen formation is suppressed by the proinflammatory cytokines TNF alpha and IFN gamma, and that these interactions are influenced by dexamethasone. Proinflammatory cytokines interact in a complex manner with other serum factors to modulate IGF-I stimulated extracellular matrix production and may have important roles in regulating tissue repair. PMID- 7490533 TI - Regulation of glucagon-like peptide-1 secretion from rat ileum by neurotransmitters and peptides. AB - Food ingestion induces a rapid increase in the insulinotropic glucagon-like peptide-1 (GLP-1) in plasma. Paradoxically, GLP-1 originates from the lower intestines and therefore a complex regulation of postprandial GLP-1 secretion must exist. This was addressed in the present study by utilizing an isolated vascularly perfused rat ileum preparation. Peptides and neurotransmitters thought to be candidate mediators triggering GLP-1 secretion were arterially infused and GLP-1 was measured in the venous effluent. Arterial infusion of cholinergic agonists strongly enhanced GLP-1 secretion which was counteracted by the addition of atropine. Histamine, dopamine, 5-hydoxytryptamine, gamma-aminobutyric acid, and norepinephrine had no effect. Peptides of the bombesin family were strong stimulants whereas tachykinins, enkephalins, dynorphin, TRH, calcitonin-gene related peptide and members of the secretin family, vasoactive intestinal peptide, peptide histidine isoleucine and neuropeptide Y, were less effective. The second incretin hormone, gastric inhibitory polypeptide (GIP), was the most potent stimulant of GLP-1 secretion in our study. It enhanced GLP-1 release up to sixfold above basal during the early phase followed by a sustained secretion at 400% above basal. This stimulation remained unaffected by atropine. In conclusion, in addition to luminal stimulation of nutrients, a cholinergic impulse as well as peptidergic mediators (among them possibly GIP and GRP) may have an impact on postprandial GLP-1 secretion from the rat ileum. PMID- 7490534 TI - Glucose activation of acetyl-CoA carboxylase in association with insulin secretion in a pancreatic beta-cell line. AB - Malonyl-CoA, which is the unique product of acetyl-CoA carboxylase (ACC), may serve as a metabolic coupler in glucose-stimulated insulin secretion by pancreatic beta-cells. Therefore we examined if and how ACC is affected by glucose in association with insulin secretion. Glucose induces a rapid increase in ACC activity which is closely related to insulin secretion in a dose- and time dependent manner. The acute effect of glucose in increasing ACC activity is caused by dephosphorylation of existing ACC molecules, without the production of new enzyme. Inhibition of ACC dephosphorylation and activation by the use of okadaic acid led to diminished glucose-mediated insulin secretion. Likewise, when ACC phosphorylation and inactivation were induced by the use of 5-amino 4 imidazole-carboxamide ribotide, an AMP analog and activator of 5'-AMP protein kinase, the glucose-induced insulin secretion was virtually nil. In the long term, glucose induced ACC and increased insulin secretion. In beta-cells, ACC gene expression is controlled by promoter II and glucose activated promoter II expression. ACC promoter I is not expressed in beta-cells. Maximum activation of ACC and insulin secretion by glucose in the short term occurred at 5 mM glucose. On the other hand, activation of the expression of ACC promoter II occurred when the cells were exposed to high glucose concentrations for a long period of time. Thus, we have shown that ACC, the only enzyme that produces malonyl-CoA, is activated by glucose; activation of ACC is accomplished by dephosphorylation in the short term and by induction of ACC by gene activation in the long term. PMID- 7490535 TI - Mechanisms of altered LH secretion in neonatally oestrogenized male rats. AB - It is well known that the control of LH secretion depends on the steroid milieu during the postnatal period. In this study LH secretion was analysed in adult male rats injected neonatally with 500 micrograms oestradiol benzoate (1) after orchidectomy, (2) after selective elimination of androgens by destruction of Leydig cells with ethylene dimethane sulphonate (EDS), and (3) after removal in orchidectomized animals of Silastic capsules containing testosterone. In addition, (4) in vivo and in vitro LH secretion in response to LHRH agonist and antagonists, (5) the hypothalamic LHRH content, (6) the basal and stimulated in vitro LHRH release, and (7) the LH responses after administration of naloxone (2 mg/kg), alpha-methyl-p-tyrosine (alpha-MPT; 250 mg/kg), N-methyl-D-aspartic acid (NMDA, 15 mg/kg) or kainic acid (KA; 15 mg/kg) were also examined. Our data indicated that (1) the LH response after orchidectomy, after EDS administration and after removal of Silastic capsules containing testosterone was diminished in oestrogenized male rats, (2) the pituitaries from oestrogenized males retained responsiveness to LHRH, (3) hypothalamic LHRH content was reduced in oestrogenized males, but the hypothalamus from oestrogenized males released more LHRH than those of control groups both under basal conditions or after depolarization, (4) alpha-MPT decreased LH secretion only in oestrogenized males, and (5) NMDA and KA stimulated LH only in oestrogenized males. We conclude that in oestrogenized male rates the loss of sensitivity to the negative feedback action of testosterone on LH secretion was not due to decreased pituitary responsiveness to LHRH stimulation or to the inherent damage of LHRH neurones.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7490536 TI - The thymic-endocrine connection. PMID- 7490537 TI - The role of excitatory amino acid pathways in the control of pituitary function in neonatally oestrogenized male rats. AB - It is well known that in the rat neonatal manipulation of the sex steroid environment results in altered hypothalamic-pituitary function in adulthood which implies an abnormal prolactin secretion and gonadotrophin response to orchidectomy. The present paper analyses the involvement of excitatory amino acid pathways in the disturbed gonadotrophin and prolactin secretion in male rats neonatally injected with oestradiol benzoate (500 micrograms on the day of birth). In the first experiment, 60-day-old control and oestrogenized male rats (intact or orchidectomized a week before) were killed 15 min after injection of vehicle, N-methyl-D-aspartic acid (NMDA; 15 mg/kg) or kainic acid (KA; 15 mg/kg). In the second experiment, prepubertal males were killed 15 min after injection of vehicle or NMDA. In the third experiment 30-, 45-, 60- and 90-day-old intact control and oestrogenized males were killed 15 min after injection of vehicle or KA. In the fourth experiment, control and oestrogenized males were sham orchidectomized, orchidectomized or orchidectomized and implanted with Silastic capsules at 30 days of age and killed on day 90, 15 min after vehicle or KA injection.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7490538 TI - Similarities between hamster pancreatic islet beta (HIT) cell vasopressin receptors and V1b receptors. AB - Vasopressin (VP) elicits almost identical insulin-stimulatory dose responses in isolated mouse islets and hamster beta (HIT) cells. We have further pharmacologically characterized HIT cell VP receptors by comparing the potencies of a series of VP agonists including the novel V1b agonist, desamino(D-3-(3' pyridyl)-Ala2,Arg8)VP (d(D-3-Pal)VP), in stimulating insulin secretion and inositol phosphate (IP) production. The relative orders of potency of VP analogues were parallel in both respects: desamino-Arg-VP (dAVP) > Arg-vasotocin (AVT) = VP > oxytocin (OXY) > desamino-D-Arg-VP (dDAVP) > d(D-3-Pal)VP. dAVP, the most potent agonist tested, behaved as a V1 but non-V1a agonist. The potency of d(D-3-Pal)VP relative to VP was 1:134 in stimulating insulin secretion and 1:40 with respect to IP production. In HIT cell monolayers, the relative order of affinity of analogues in competition for binding with [3H]AVP was: dAVP > AVT = VP > V1a antagonist > OXY > dDAVP > V2 antagonist = d(D-3-Pal)VP, in parallel with their biological activity. The relative orders of potency and affinity parallel those reported for corticotrophic V1b receptors. Binding studies with hamster liver membranes indicate that the hepatic VP receptor belongs to the V1a class. We conclude that VP activates phospholipase C and interacts with functional VP receptors of the V1 type, which do not belong to the V1a subclass and which are similar to V1b receptors. PMID- 7490541 TI - Glucagon improves insulin secretion from pig islets in vitro. AB - It has been shown that peripheral glucagon secreting cells (A-cells) are lost during most of the isolation procedures employed for pig islets. Loss of A-cells decreases intra-islet glucagon levels and cAMP levels in B-cells and might reduce glucose-induced insulin release. This study was designed to test this hypothesis, by evaluating the effects of culture of porcine islets with exogenous glucagon on insulin secretion and on insulin and cAMP content in islets. Islets were isolated from adult 2-year old Large White pigs using an automated method. The number of A cells was calculated by immunostaining for glucagon in islets before and after isolation and a significant decrease in A-cells was observed. After an overnight culture, islets were cultured for 48 h in a standard medium (CMRL 1066, 10% foetal calf serum, 1% antibiotics, 1% glutamine) alone or in the presence of glucagon at two different concentrations (1.0 and 10.0 microM); exposure to glucagon was either continuous or alternated with periods of incubation in CMRL 1066 alone. After the 48-h culture in standard medium, the islet glucagon response to arginine was almost negligible and significantly lower than that observed in human islets.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7490539 TI - Synergism of GH and IGF-I in stimulation of sulphate uptake by teleostean branchial cartilage in vitro. AB - The combined effects of teleostean GH and human IGF-I in the regulation of teleost branchial cartilage growth were examined. Ceratobranchial cartilages were dissected from the common carp (Cyprinus carpio) and maintained in a defined culture medium supplemented with recombinant striped bass GH (sbGH), human IFG-I (hIGF-I) or both, and the uptake of [35S]sulphate by cartilage explants was measured. sbGH alone in the culture medium did not exhibit a significant stimulatory effect on the uptake of [35S]sulphate when compared with the controls. However, with hIGF-I in the culture medium, as low as 1 ng/ml, the stimulatory effect of sbGH was apparent and dose-dependent. The synergism of sbGH and hIGF-I was observed at the concentrations of 1 and 10 ng IGF-I/ml tested. At a constant hIGF-I concentration (10 ng/ml), a maximum stimulatory effect was detected with 3 micrograms recombinant sbGH/ml, at which point a 2.3-fold increase in sulphation activity was obtained when compared with the treatment with hIGF-I alone. A similar dose-dependent stimulatory effect was observed when native common carp and bonito GHs were tested using this assay system. These results suggest that the synergistic effect of sbGH and hIGF-I on sulphation activity is a common biological function of teleost GH, thus ruling out the possibility of experimental artifacts resulting from utilization of heterologous GH. Furthermore, experiments were conducted to test whether the synergism between sbGH and hIGF-I occurred between sbGH and insulin.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7490540 TI - The effect of nutrition on testicular growth in mature Merino rams involves mechanisms that are independent of changes in GnRH pulse frequency. AB - In mature Merino rams, changes in diet to below or above the requirements for maintenance of body weight lead to changes in gonadotrophin secretion and testicular growth. However, the effects on testicular growth persist for much longer than those on LH and FSH secretion so that the gonadal and gonadotrophin responses are poorly correlated over time. This suggests that the gonadal effects may be partly independent of changes in the hypothalamic secretion of GnRH, an hypothesis tested in this study. In a short-term experiment (November, late spring, non-breeding season), we tested whether a high frequency of exogenous GnRH pulses could override the endogenous system and mimic the change in gonadotrophins seen in rams fed a high plane of nutrition. Mature Merino rams (scrotal circumference (mean +/- S.E.M.) 33.6 +/- 0.5 cm, body weight (mean +/- S.E.M.) 59.0 +/- 0.9 kg) were fed 900 g chaff + 1.6 kg lupin grain (High diet) or 360 g chaff + 60 g lupin grain (Low diet) and infused with 8 pulses of GnRH or saline daily for 5 weeks (n = 5/group). Blood was sampled every 20 min for 12 h on days-1 and 14 relative to the start of treatments. Relative to pre-treatment levels, LH pulse frequency and FSH concentrations were decreased on day 14 in saline-infused rams fed the Low diet and increased in saline-infused rams fed the High diet (P < 0.001). In GnRH-infused rams, gonadotrophin secretion was not affected by diet and the patterns of secretion of LH and FSH were similar to those in saline-infused rams fed the High diet. This model was used for a more complete endocrine analysis in a longer experiment designed to test the hypothesis that the effect of nutrition on testicular growth is partly independent of changes in the secretion of GnRH. The same treatments were imposed for 35 days on a different group of similar rams in March (autumn, mid-breeding season). Body weight and scrotal circumference were measured weekly and blood was sampled on days -1 and 14. On days -1 and 35, testosterone secretion in response to LH was tested by injecting exogenous ovine LH (NIADDK-oLH-25; 200 ng/kg body weight) to all rams. Body weight increased in rams fed the High diet and decreased in those fed the Low diet (P < 0.001) and was not affected by treatments as in experiment 1.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 7490542 TI - Effect of octreotide on experimental diabetic renal and glomerular growth: importance of early intervention. AB - IGF-I acts as a renotropic factor in early streptozotocin-induced diabetes. Somatostatin analogue (octreotide) treatment initiated at the onset of diabetes prevents kidney IGF-I accumulation and renal growth. Seven days of octreotide treatment initiated after 3, 5, 7 or 9 days of untreated diabetes was investigated. Diabetic renal hypertrophy was followed by renal hyperplasia. Compared with placebo-treated diabetic rats, the earliest octreotide intervention was followed by a greater reduction in renal growth compared with intervention later on (days 3 to 10, 12%; days 5 to 12, 10%; days 7 to 14, 9%; days 9 to 16, 6%; P < 0.05). Octreotide treatment was unable to reduce protein accumulation and kidney DNA increase consistently. No difference in glomerular volume fraction or total glomerular volume was observed between placebo- and octreotide-treated diabetic rats. Octreotide treatment was followed by reduced kidney and serum IGF I especially following early intervention, while no effect over that of diabetes was observed in the later intervention periods. The results confirm the notion that initial renal IGF-I accumulation is a prerequisite for early diabetic kidney hypertrophy in rats and show that delayed octreotide treatment cannot reverse renal and glomerular growth which is already manifest. PMID- 7490543 TI - Agouti protein can act independently of melanocyte-stimulating hormone to inhibit melanogenesis. AB - In animals, the coat-darkening effects of alpha-melanocyte stimulating hormone (alpha-MSH) are opposed by agouti protein. Although agouti protein has been shown to be a competitive antagonist of the melanocyte-associated MC-1 melanocortin receptor, the possibility that agouti protein can affect melanogenesis independently of its ability to antagonise melanocortin activity cannot be excluded. This study demonstrates that murine agouti protein causes both a time- and concentration-dependent suppression of melanogenesis in B16 F1 murine melanoma cells. In addition, human agouti protein decreases melanogenesis in cultured human epidermal melanocytes. However, agouti protein has little effect on the ability of alpha-MSH to stimulate melanogenesis. These observations raise fundamental questions about the mode of action of agouti protein in regulating melanogenesis. PMID- 7490544 TI - Effects of maternal diabetes on fetal expression of insulin-like growth factor and insulin-like growth factor binding protein mRNAs in the rat. AB - Maternal diabetes is associated in humans and rats with an increased risk for fetal growth abnormalities and malformations. Therefore, the effect of maternal diabetes on expression of genes that regulate fetal growth and differentiation is of considerable interest. Developmental growth is regulated in part by the expression and availability of insulin-like growth factors (IGFs). Postnatal expression of a subset of the IGFs and IGF binding proteins (IGFBPs) has been demonstrated to be regulated in response to diabetes and other metabolic conditions. We used in situ hybridization to analyze the effect of maternal diabetes, induced by streptozotocin (STZ) prior to mating, upon prenatal rat IGF and IGFBP mRNA expression. At gestational day (GD) 14, the most striking effect of maternal diabetes on fetal IGF/IGFBP gene expression was a marked increase in the abundance of IGFBP-1 mRNA within the liver primordia of fetuses isolated from diabetic dams compared to age-matched controls. This upregulation cannot be entirely due to the approximately one-half-day delay in fetal development (based on limb bud staging) associated with maternal diabetes, as there was no gross difference in the level of IGFBP-1 mRNA between GD13 and GD14 control fetal livers. In contrast, the fetal mRNA expression patterns of IGF-I, IGF-II and IGFBP-2, -3, -4, -5 and -6 were not grossly altered by maternal diabetes. These data are consistent with the hypothesis that IGFBP-1 produced within the fetal liver and secreted into fetal circulation may play a role in regulating rat fetal growth. PMID- 7490545 TI - IL-6sR: release from MCF-7 breast cancer cells and role in regulating peripheral oestrogen synthesis. AB - The interleukin-6 soluble receptor (IL-6sR) may regulate the ability of IL-6 to stimulate oestrogen synthesis in breast cancer cells and breast tumours. Significant aromatase activity was detectable in IL-6 stimulated fibroblasts derived from subcutaneous adipose tissue, but the combination of IL-6sR plus IL-6 resulted in a marked 21-fold stimulation of aromatase activity. To examine the control of IL-6sR release, the effects of oestradiol, 4-hydroxytamoxifen (4-OHT), dexamethasone, TPA, TNF alpha or IL-6 on this process was examined using MCF-7 breast cancer cells. Oestradiol, TNF alpha and dexamethasone all markedly increased IL-6sR release. While 4-OHT had a small stimulatory effect on IL-6sR release, it blocked the ability of oestradiol to increase IL-6sR release. Significant concentrations of IL-6sR were also detected in conditioned medium collected from lymphocytes and macrophages and in cytosols prepared from normal and malignant breast tissues. These results indicate that IL-6sR may have an important role in potentiating the effect of IL-6 on oestrogen synthesis in breast cancer cells. The abilities of oestradiol or tamoxifen to potentiate or inhibit the IL-6 stimulation of oestrogen synthesis in breast cancer cells may result from their effects on IL-6sR release. PMID- 7490546 TI - Genetics of XY sex reversal. PMID- 7490547 TI - Endocrine signals from the mammary gland. PMID- 7490549 TI - Type I insulin-like growth factor receptor expression in different developmental stages of human thymocytes. AB - Insulin-like growth factor-I (IGF-I) has been implicated in playing a regulatory role in T cell development and in T cell function. We demonstrate the presence of type I IGF receptors on human thymocytes using radioligand binding assays and flowcytometric analysis. The relative potencies of IGF-I, IGF-II and insulin for competition with 125I- IGF-I indicate the presence of type I IGF receptors. Scatchard analysis revealed that the average number of receptors per thymocyte is 257 +/- 28 with a Kd of 0.12 +/- 0.01. With multicolour flowcytometry using a type I IGF receptor specific monoclonal antibody (alpha IR3), we show that CD4 CD8- cells express 3-4 times more receptors per cell as compared with CD4+CD8+, CD4+CD8- and CD4-CD8+ cells. IGF-I directly stimulated DNA synthesis of thymocytes and potentiated DNA synthesis in mitogen-activated thymocytes. These results indicate that IGF-I can influence T cell development in humans at the level of the thymus. PMID- 7490548 TI - Effects of gonadal steroids on the opioid regulation of LH and prolactin release in ovariectomized pony mares. AB - The aim of the present study was to investigate the role of ovarian steroids in the opioid regulation of LH and prolactin release in mares. Effects of the opioid antagonist naloxone on LH and prolactin secretion were determined in ovariectomized pony mares. The animals were pretreated with either progesterone (500 micrograms kg-1) or oestradiol benzoate (10 micrograms kg-1) for 8 days and subsequently with a combination of progesterone and oestradiol for an additional 8 days. Naloxone administration (0.5 mg kg-1 i.v.) resulted in a significant release of LH as well as prolactin in mares after pretreatment with either oestradiol benzoate or progesterone plus oestradiol benzoate (P < 0.05). No significant changes in LH and prolactin secretion were detected in progesterone treated and non-steroid-treated ovariectomized mares. These results indicate that a prolonged oestrogen influence activates the opioid inhibition of LH and prolactin release in mares. In contrast to other species, progesterone alone does not activate a tonic opioid inhibition of LH and prolactin secretion, but modulates the effect of oestrogens. The opioid systems therefore seem to be regulated by a sequence of different steroid environments, as found during the oestrous cycle. The parallel increases in prolactin and LH secretion in mares may indicate a common regulatory pathway for these two hormones. PMID- 7490550 TI - Urinary clearance of atrial natriuretic peptide in Pekin ducks. AB - To quantify the renal excretion of atrial natriuretic peptide (ANP) in birds, synthetic ANP was infused intravenously into conscious Pekin ducks at rates of 10, 30 and 90 ng/kg per min for 30 min and the relationships between plasma and urine ANP concentrations were monitored by RIA. The infusions of ANP produced dose-dependent elevations in the plasma concentrations of the hormone; however, urinary concentrations and excretion rates did not change significantly from the basal values of 142.2 +/- 21.5 pg/ml and 26.4 +/- 4.5 pg/min per kg respectively. The metabolic clearance rate of ANP remained constant at each infusion dose (approximately 200 ml/min per kg). The urinary clearance rates (< 0.3 ml/min per kg) indicated that the fraction of ANP cleared from the blood via the urine was less than 0.2%. Clearly, excretion of intact ANP via this route is not a quantitatively important means of elimination for this hormone in birds. PMID- 7490551 TI - Pituitary and plasma levels of growth hormone in Booroola sheep that are either homozygous carriers or non-carriers of the FecBB fecundity gene. AB - The aim of this study was to examine the effect of the FecBB fecundity gene on plasma concentrations and pituitary content of growth hormone (GH) in sheep. No differences were found between homozygous carriers (BB) and non carriers (++) of the FecBB gene with regard to pituitary GH contents in both ovariectomized and intact ewes. However, ovariectomized ewes had higher levels of pituitary GH than intact ewes (P < 0.01). There were no differences between FecBB genotypes with respect to plasma concentrations of GH in 6-year-old ovariectomized ewes bled every 10 min for 12 h or in ram lambs bled weekly during their first year of life. GH levels in the rams decreased until week 27, increased to a peak at week 31 then decreased before increasing again at week 43. Mean plasma GH concentrations in the ewe lambs bled weekly for a year decreased until week 19 then remained at approximately this level for the remainder of the year. Mean GH plasma concentrations in the ram lambs were higher than in the ewe lambs (P < 0.001). Ewe lambs that were homozygous for the FecBB gene had lower body weights (P < 0.05) and had higher levels of GH (P < 0.01) than non carrier ewe lambs during their first year. Before the average age of first behavioural oestrus (36 weeks) GH levels in the ewe lambs were negatively correlated with body weights (r = -0.69, P < 0.001, n = 22). When body weight was included as a covariate in analysis of variance the genotype difference in ewe lamb plasma GH concentrations was no longer significant.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7490552 TI - Hypothalamic GH receptor gene expression in the rat: effects of altered GH status. AB - GH synthesis and release from the anterior pituitary is governed by the opposing actions of somatostatin (SS) and GH-releasing factor (GRF), derived from the periventricular and arcuate nucleus (ARC) of the hypothalamus respectively. GH is known to regulate its own release by hypothalamic autofeedback mechanisms, but the extent to which this is a direct effect rather than indirectly via the generation of IGFs is still a subject of debate. GH receptors are known to be present in the hypothalamus, but their physiological regulation is poorly understood. We therefore used in situ hybridization histochemistry to investigate the effects of GH status on hypothalamic GH receptor gene expression, using hypophysectomized normal and dw/dw dwarf rats as models of acquired and congenital GH deficiency. Hypophysectomy resulted in a time-dependent reduction in GH receptor gene expression. ARC GH receptor transcripts in untreated dw/dw dwarf rats were half those found in normal animals of the same background strain (16.8 +/- 1.7 vs 9.3 +/- 1.9 d.p.m./mg, P < 0.05). Increasing circulating GH by peripheral infusion of 200 micrograms human GH (hGH)/day for 6 days increased ARC GH receptor expression in dw/dw rats to normal. In contrast, central infusions of hGH at 26.4 and 79.2 micrograms/day for 6 days in normal rats lowered ARC GH receptor gene expression. The sensitivity of GH receptor gene expression within the central nervous system to peripheral and central GH levels suggests that feedback regulation of GRF and/or SS may be mediated directly by these receptors, and that the sensitivity to GH feedback is also subject to autoregulation by GH altering its own receptor expression. PMID- 7490553 TI - Endothelin-1 and endothelin receptors are present in the sheep uterus and conceptus at implantation. AB - Previous studies have demonstrated that endothelin is present in the ovine endometrium and increases at around the expected time of implantation. To characterize further uterine endothelin at the time of establishment of pregnancy in sheep, endothelin was measured by radioimmunoassay in uterine flushings obtained during the oestrous cycle and in pregnant ewes up to the time of implantation (day 16). During the oestrous cycle, the highest amounts of endothelin were present in uterine flushings on day 14 (1.1 +/- 0.2 ng endothelin/uterus). During early pregnancy, basal levels of endothelin (0.5-0.6 ng endothelin/uterus) were present in uterine flushings for the first 10 days and then increased on day 14 to levels similar to those found at the equivalent stage of the oestrous cycle. On days 15 and 16 of pregnancy, endothelin content in the uterine lumen increased to significantly (P < 0.05) higher concentrations (2.9 +/ 0.4 ng endothelin/uterus) when compared with the non-fertile cycle. The principal isoform present in flushings at the time of implantation was endothelin 1, as determined by reverse-phase HPLC. Endothelin was released principally by purified endometrial epithelial cells in culture, with barely detectable amounts released by endometrial stromal cells or conceptus tissue, which is consistent with the epithelium being the principal source of endothelin in the uterine lumen. Endothelin binding sites were present in endometrium and myometrium, as demonstrated by specific binding of 125I-labelled endothelin-1, which was saturable and displaced by endothelin-1. Both endothelinA and B sub-types of receptors were present as demonstrated by the biphasic displacement of 125I labelled endothelin-1 binding by the specific endothelinB agonist BQ3020. These were localised principally on luminal and glandular epithelium and in the vasculature of the endometrium and myometrium as shown by autoradiography. Endothelin receptors were also present on the conceptus obtained at the time of implantation. In the day 20 conceptus, endothelin immunostaining was localised principally in the heart, in trophoblast in uninucleate but not in binucleate cells, and in fetal membranes. This immunostaining of the conceptus may represent binding to receptor sites. It is concluded that endothelin-1 is present in the uterine lumen and may play an important role in the paracrine regulation of the conceptus and endometrium at the time of rapid embryo development, implantation and early placentation. PMID- 7490554 TI - Amniotic and allantoic fluid concentrations of thyroxine, 3,3',5'-tri iodothyronine, 3,3'-di-iodothyronine and 3',5'-di-iodothyronine in the pig during the period of gestation. AB - Thyroxine (T4), 3,3',5'-tri-iodothyronine (reverse T3; rT3) and di-iodothyronines (3,3'-T2 and 3',5'-T2) were measured in pig amniotic fluid (AF) and allantoic fluid (Al) between 32 and 113 days of normal pregnancy. Low but measurable quantities of T4 in AF and Al (2.1 +/- 0.3 and 3.2 +/- 0.5 nmol/l respectively) were found before the onset of fetal thyroid gland function, which indicates the maternal source of T4. The presence of rT3 (55.8 +/- 4.1 pmol/l in AF and 49.8 +/ 5.3 pmol/l in Al), 3,3'-T2 (45.5 +/- 0.6 pmol/l in AF and 49.2 +/- 9.2 pmol/l in Al) and 3',5'-T2 (20.8 +/- 2.6 pmol/l in AF and 24.0 +/- 2.2 pmol/l in Al) may be attributed to the monodeiodinase system already active in fetal pig tissues in early pregnancy, as demonstrated previously. T3 concentration was undetectable in both AF and Al. An approximately twofold increase in the levels of T4, rT3 and T2s in AF and Al at mid-gestation was observed. T4 and rT3 in AF showed a positive correlation with protein concentrations. AF rT3 concentration (but not T4) correlated with rT3 in the cord and maternal serum. The 3,3'-T2 and 3',5'-T2 in AF and Al showed parallel changes to rT3, while the rT3/3,3'-T2 and rT3/3',5' T2 molar ratios remained constant. T4 concentrations in AF and Al were markedly lower than in corresponding maternal and fetal serum; the rT3 concentration in Al was equal to that in AF and two to four times lower than in fetal serum. In spite of differences between serum hormone patterns in the pig and human near term, iodothyronine concentrations in AF showed some similarities, mainly the following: undetectable T3, a strong correlation between rT3, T4 and AF total protein and the presence of 3,3'-T2 and 3',5'-T2 in measurable levels. Comparative data for Al, except the ones in the present study in the pig, are not available. PMID- 7490555 TI - Growth hormone (GH) receptor and GH-binding protein deficiency in the growth failure of potassium-depleted rats. AB - Potassium (K+) deficiency is associated with growth retardation in both man and experimental animals. Growth hormone (GH) administration to such animals prevents, to some extent, weight loss and selective muscle atrophy, but does not affect tail and tibia length even with supraphysiological doses. The present study was undertaken to investigate the possible effect of K+ deficiency on the hepatic GH receptor and GH-binding protein (BP). Young female Wistar rats were maintained on K(+)-deficient fodder and distilled water, and compared with pair fed and ad-libitum-fed control groups. After 15 days GH-BP and electrolytes were measured in sera, GH receptors were studied in liver membranes by 125I-labeled human GH binding and muscles were weighed and saved for electrolyte measurements. K(+)-deficient rats showed complete growth arrest compared with an intermediate weight gain of the pair-fed group. Serum K+ was very low, at 1.5 +/- 0.1 mmol/l, compared with the mean value of 5.3 mmol/l of control animals. Somatogenic and lactogenic receptors in liver membranes and serum GH-BP levels were significantly (P < 0.05) lower in K+ deficiency, as compared with their pair-fed controls. Liver GH receptors correlated significantly (P < 0.05) with serum GH-BP levels. The growth variables correlated positively with both hepatic somatogenic and lactogenic receptors and serum GH-BP levels, with correlation coefficients that were highest against serum GH-BP and lowest against liver lactogenic receptors. Serum and muscle K+ correlated significantly (P < 0.05) with both liver GH receptors and serum GH-BP, with correlation coefficients that were higher against serum GH-BP.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7490556 TI - Mobilization of LH secretory granules in gonadotrophs in relation to gene expression, synthesis and secretion of LH during the preovulatory phase of the sheep oestrous cycle. AB - Changes in LH secretory granules in pituitary gonadotrophs throughout the sheep oestrous cycle were determined by immunogold localisation of LH at ultrastructural level by electron microscopy. Oestrous cycles in Welsh Mountain ewes were initially synchronised with progestagen sponges and studies carried out in the subsequent cycle. Animals were allocated at random to six groups each of five animals, one killed on day 12 of the luteal phase and the other groups after prostaglandin (PG)-induced luteal regression at PG plus 18 h (early follicular phase), oestrus (PG plus 33.6 +/- 1.0 h), oestrus plus 9 h just before the preovulatory LH surge, 1 h after GnRH agonist-induced LH surge at PG plus 48 h (mid-LH surge) and oestrus plus 24 h, after the preovulatory LH surge. Blood samples collected throughout confirmed the pulsatile secretion of LH before and the timing in relation to the preovulatory LH surge. Pituitaries were dissected and processed for transmission electron microscopy and frozen for later extraction of mRNA. Only a single type of LH cell was present in the sheep pituitary. In the luteal phase, LH-immunopositive secretory granules were distributed throughout the cytoplasm in 80% of cells while in 20% of cells granules were polarised to the region of the cell next to a vascular sinusoid. The percentage of polarised cells increased during the follicular phase to 45% at oestrus, 75% at oestrus plus 9 h just before the LH surge and 90% in mid-LH surge. Cell size increased in parallel with polarisation. Gonadotrophs after the LH surge were almost totally devoid of LH granules but prominent LH beta immunoreactivity was observed in the rough endoplasmic reticulum. Analysis of granule diameters revealed a single class of granules with a maximum diameter of 300 nm. Polarised cells had significantly fewer 130-150 nm granules than non polarised cells, suggesting preferential exocytosis of LH-containing granules of this size from polarised cells. Northern analysis showed that LH beta mRNA levels decreased from luteal through the follicular phase. These results suggest that the preovulatory LH surge in sheep is not related to a change in synthesis of LH but to a progressive recruitment of gonadotrophs into a releasing state, priming, as indicated by polarisation of secretory granules to the region of the cell next to the vascular system. PMID- 7490557 TI - Changes in inhibin-A (alpha-beta A dimer) and total alpha inhibin in the peripheral circulation and ovaries of rats after gonadotrophin-induced follicular development and during the normal oestrous cycle. AB - Recent modifications to a previously reported two-site IRMA have permitted the measurement of serum/plasma concentrations and ovarian contents of inhibin-A (alpha-beta A dimer) in pregnant mare serum gonadotrophin (PMSG)-treated immature female rats and adult rats throughout the 4-day oestrous cycle. For comparison, total alpha inhibin levels were also measured by alpha subunit-directed inhibin RIA and found to be at least tenfold higher (relative to the same 32 kDa bovine inhibin standard used to calibrate both assays). In immature female rats, serum levels of inhibin-A dimer and total alpha inhibin increased within 3 h of PMSG injection and rose in parallel over the next 48 h to values four- to fivefold higher than pretreatment levels. Ovariectomy led to a rapid and parallel fall in both inhibin-A dimer and total alpha inhibin; initial half-lives (+/- 95% confidence intervals) were 22 +/- 4 and 20 +/- 5 min respectively. In adult rats, marked fluctuations in plasma concentrations and ovarian contents of inhibin-A dimer and total alpha inhibin occurred during the 4-day oestrous cycle, most notably between the morning of pro-oestrus and the morning of oestrus. Plasma levels of inhibin-A dimer and total alpha inhibin peaked on the afternoon of pro oestrus just before the preovulatory gonadotrophin surge. After ovulation, both inhibin-A dimer and total alpha inhibin fell abruptly (two- to threefold by 0200 h on oestrus; P < 0.001), while FSH showed a secondary rise which peaked at 0700 h on oestrus. Although IRMA- and RIA-derived inhibin values generally followed a similar pattern across the 4-day cycle (plasma: r = 0.52, P < 0.001; ovary: r = 0.41, P < 0.001), a transient rise in plasma and ovarian inhibin-A dimer was detected at 0700 h on oestrus (P < 0.01) which was unaccompanied by a rise in total alpha inhibin. This rise in plasma inhibin-A dimer was probably responsible for terminating the post-ovulatory FSH surge since FSH levels declined steadily over the next 15 h. Overall, plasma inhibin-A dimer and FSH concentrations across the whole cycle were negatively correlated (r = -0.22, P < 0.01) whereas no correlation existed between total alpha inhibin and FSH (r = -0.11, P = 0.12). PMID- 7490559 TI - Inhibition of 5 alpha-reductase in genital skin fibroblasts and prostate tissue by dietary lignans and isoflavonoids. AB - Isoflavonoids and lignans, constituents of many plant foods, have been proposed as protective agents in those populations with a low incidence of hormone dependent cancers. They may act by their inhibition of the metabolism of growth promoting steroid hormones. This report describes the inhibition of 5 alpha reductase and 17 beta-hydroxysteroid dehydrogenase by six isoflavonoids and two lignans in human genital skin fibroblast monolayers and homogenates, and in benign prostatic hyperplasia tissue homogenates. In genital skin fibroblasts, genistein, biochanin A and equol were the most potent inhibitors of 5 alpha reductase activity, each resulting in greater than 80% inhibition at a concentration of 100 microM. The IC50 values for genistein and a seven-compound mixture were approximately 35 microM and 20 microM (2.9 microM of each compound) respectively. Of the lignans, enterolactone was the most potent inhibitor. Inhibition by biochanin A was shown to be reversible. When genital skin fibroblast homogenates were used, biochanin A was found to inhibit 5 alpha reductase isozymes 1 and 2 to differing extents (30% and 75% respectively). Genistein was shown to inhibit 5 alpha-reductase 2 in a non-competitive nature (Vmax and Km values without and with inhibitor were 30 and 20 pmol/mg protein per h and 177 and 170 nM respectively). All of the compounds tested inhibited 17 beta hydroxysteroid dehydrogenase activity in genital skin fibroblast monolayers. When prostate tissue homogenates were used, the compounds tested were better inhibitors of 5 alpha-reductase 1 than 2.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7490558 TI - Immunolocalisation of androgen receptor to interstitial cells in fetal rat testes and to mesenchymal and epithelial cells of associated ducts. AB - Androgens are required for the development of male internal and external genitalia. Androgen action is mediated by an intracellular receptor which acts as a transcription factor following activation by ligand binding. The aim of the present study was to define the time of appearance of androgen receptor (AR) in the male fetal rat gonad using immunohistochemistry. Intact fetuses (days 13.5 16.5) or testicular tissue (days 16.5-20.5 and days 3-7 postnatal) were fixed in Bouins' solution and processed into paraffin wax. On day 16.5 nuclear AR were present in mesenchymal cells surrounding the Wolffian duct but those around the Mullerian duct were receptor negative. During the following day (17-18) the abundance of nuclear staining increased, becoming detectable in the epithelial cells of the Wolffian ducts. Within the testis some nuclear staining was apparent at day 17 but was confined to interstitial cells surrounding the seminiferous cords. As development of the testis proceeded the abundance of nuclear AR in peritubular and elongated mesenchymal cells increased. AR were not detected in fetal Leydig cells expressing 3 beta-hydroxysteroid dehydrogenase nor in the ovaries or associated ducts of female fetuses at the same ages. In conclusion, in the rat we have found AR expression detectable by immunohistochemistry in mesonephric mesenchyme to be confined to that underlying the Wolffian ducts and to be absent from the area around the degenerating Mullerian duct. On and after day 17 of gestation AR is present in Wolffian duct epithelial cell nuclei and within the testis it is confined to peritubular and interstitial cells which may have migrated from the mesonephros.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7490560 TI - Multiple mechanisms of heterologous beta-adrenoceptor regulation in rat uterus. AB - The inhibitory effects of catecholamines on rat myometrium mediated by beta adrenoceptors are modulated by ovarian steroids. Previously reported findings of radioligand binding studies on myometrial membranes have demonstrated changes in the numbers of beta-adrenergic binding sites following ovarian steroid treatment. However, these changes were not accompanied by parallel functional changes. In the present study, we have investigated possible mechanisms of heterologous beta adrenoceptor regulation by ovarian steroids. Binding studies were performed on myometrial membrane and cytosolic preparations from rats which had been ovariectomized and subsequently received no hormonal treatment or had been treated with oestradiol, progesterone or combined oestradiol and progesterone. The beta-adrenergic antagonist [125I]iodocyanopindolol and the unlabelled competing agonist, isoprenaline, were used in the present studies. Hormonal treatment had no effect on the concentration of beta-adrenergic binding sites in the myometrium (i.e. the number of membrane-bound and cytosolic binding sites per mg protein). However, significant changes were found in the total number of binding sites; these were associated with the hormone-induced tissue hypertrophy. In myometrium from ovariectomized-alone rats, approximately 50% of beta adrenergic binding sites were present in the cytosolic fraction. Oestradiol treatment, either on its own or in combination with progesterone, resulted in the translocation of binding sites to the cell membrane. However, in the absence of progesterone only 33% of the membrane-bound binding sites bound the beta adrenergic agonist, isoprenaline, with a high affinity, suggesting that the majority of these membrane-bound binding sites represented non-functional beta adrenoceptors.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7490561 TI - Two-step development of Hashimoto-like thyroiditis in genetically autoimmune prone non-obese diabetic mice: effects of iodine-induced cell necrosis. AB - The administration of a high iodide dose (HID; 10 micrograms/day) to goitrous mice is known to induce thyroid cell necrosis and inflammation, which, in most strains, is transient. In this study, we analyzed the effects of iodide in autoimmune prone non-obese diabetic (NOD) mice. Control NOD mice fed a standard diet (MID; 1 microgram I/day) or HID did not spontaneously develop thyroiditis. In NOD mice previously made goitrous, HID provoked thyroid cell necrosis and diffuse inflammation within 4 days. Inflammatory cells consisted of MHC-class II+ antigen-presenting cells, CD4+ T helper cells and CD8+ T suppressor/cytotoxic cells. After 96 days of treatment with HID, thyroiditis similar to Hashimoto's disease was obtained in 100% of the animals, with destruction of thyroid follicles, large clusters of T and B cells, and antithyroid antibodies in the plasma. When treating goitrous mice with MID, no cell necrosis was observed and no autoimmune thyroiditis was obtained. The early iodide-induced cell necrosis and inflammation may thus be considered as an important factor in the induction and persistence of autoimmune thyroiditis in individuals carrying a genetic susceptibility to autoimmune disease. PMID- 7490563 TI - Anti-progesterone effects on maternal recognition and behaviour imprinted during first pregnancy in mice. AB - Anti-progesterone treatment using specific anti-progesterone antibodies or a progesterone receptor (PR) antagonist during first pregnancy impairs postpartum maternal behaviour in mice. This effect is demonstrable only if the treatment is given during pregnancy but not immediately after parturition. The purpose of the present studies was to investigate if maternal behaviour is also impaired by anti progesterone treatment in subsequent pregnancies. Studies with a monoclonal antibody to progesterone (DB3; 4.5 nmol/mouse) showed that injection of females on day 17 of second pregnancy did not cause maternal rejection but the latency of pup retrieval was prolonged especially during the first 3 days of lactation. This phenomenon was not observed in animals that had previous experience of full length lactation. Experiments were carried out with mifepristone (RU486; 10 micrograms/mouse) injected at day 17 of first, second or third pregnancies. Pup rejection (22.5% vs 12.3%) and prolongation of the retrieval latency (62.3 +/- 13.3 vs 19.7 +/- 6.5 s; P < 0.02) were observed following the first pregnancy. No abnormal behavioural effects were found in mothers treated in second or third pregnancy who had prior full length lactation experience. Control females subjected to only one pup retrieval test after first delivery rejected their pups if treated in their second pregnancy (27.3% vs 4.4%; P < 0.001) and displayed a marginal prolongation of the retrieval latency period (20.9 +/- 7.0 vs 7.4 +/- 2.6 s). Anti-progesterone treatment had no negative influence when administered during third pregnancy.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7490562 TI - The role of intracerebral insulin in the effect of nutrition on gonadotrophin secretion in mature male sheep. AB - The effect of nutrition on gonadotrophin secretion may be exerted through a central metabolic signal that reflects nutritional status. We have previously found that glucose and insulin concentrations are elevated in the cerebrospinal fluid (CSF) of rams in which the secretion of gonadotrophins has been stimulated by a nutritional supplement of lupin grain (Lupinus angustifolius). In the present study, we tested the hypothesis that insulin and/or glucose is a metabolic modulator of GnRH secretion and mediates the effects of nutrition on gonadotrophin secretion. Six mature rams were fed a diet that maintained live weight and then given a series of infusions, each for 12 h/day for 4 days, in a cross-over design. The treatments were: artificial CSF (aCSF), glucose (50 mumol/h) in aCSF, insulin (0.6 ng/h) in aCSF, and glucose (50 mumol/h) plus insulin (0.6 ng/h) in aCSF; all infused at a rate of 5 microliters/min. At the same time as the infusion treatments, two other groups of four rams without cerebral cannulae were fed either the maintenance diet or the same diet supplemented with 750 g lupin grain per head per day for 4 days, again in a cross over design. Rams fed the lupin supplement showed an increase in both LH pulse frequency and mean FSH on day 4 (P < 0.05). Infusion of aCSF or glucose did not affect gonadotrophin secretion. Rams infused with insulin or insulin plus glucose showed an increase (P < 0.05) in LH pulse frequency but no increase in FSH concentrations on day 4 of infusion.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7490565 TI - Prolactin receptor expression in lymphocytes from patients with hyperprolactinemia or acromegaly. AB - Previous reports demonstrated that prolactin receptors (PRL-R) are widely expressed on cells of the immune system. We analyzed a possible regulation of PRL R expression on human mononucleated blood cells by prolactin (PRL) itself. PRL-R expression was analyzed by immunofluorescence on T and B lymphocytes and monocytes from peripheral blood mononucleated cells (PBMC) of patients with hyperprolactinemia or acromegaly compared with sex- and age-matched control subjects. The frequency of PRL-R positive cells and the intensity of PRL-R expression was only modified among the CD8+ T cell population of hyperprolactinemic patients with macroadenoma. No correlation was reported between PRL-R expression and circulating PRL levels. The percentage of PRL-R+ cells on B or T lymphocytes and monocytes as well as the capacity of PBMC to proliferate in response to T cell mitogens were not significantly different in bromocriptine-treated compared with untreated patients. These findings suggest that factors other than pituitary PRL play the major role in regulating PRL-R expression on cells of the immune system. PMID- 7490564 TI - Differential expression of vascular endothelial growth factor and its receptor mRNAs in the mouse uterus around the time of implantation. AB - Vascular endothelial growth factor (VEGF) is a mitogen for endothelial cells and an inducer of angiogenesis. VEGF is also known as a vascular permeability factor because it can stimulate vascular permeability. In the rodent, increased uterine vascular permeability occurs at the sites of blastocysts with the onset of the attachment reaction. This is followed by stromal decidualization and angiogenesis. We examined the temporal and spatial expression of VEGF and its receptors, Flk-1 and Flt-1, in the mouse uterus during the peri-implantation period (days 1-8) using Northern and in situ hybridization to assess the involvement of VEGF in the process of implantation. Primarily, a major (approximately 4.2 kb) transcript for VEGF mRNA was detected in uterine poly(A)+ samples, except for the presence of two other minor (approximately 3.7 and 2.5 kb) transcripts in decidual samples. The steady-state levels of these transcripts did not vary much during the peri-implantation period, except for an increase in day-8 decidual samples. Results of in situ hybridization experiments demonstrated accumulation of VEGF mRNA in the luminal epithelium on days 1 and 2. In contrast, stromal cells exhibited a modest level of signals on day 3. On day 4, luminal epithelial cells and those in the subepithelial stromal bed accumulated VEGF mRNA. On days 5-7, a clear cell type-specific accumulation of this mRNA was noted. On day 5 after the initial attachment reaction, luminal epithelial and stromal cells immediately surrounding the blastocyst exhibited accumulation of VEGF mRNA. On days 6-8, the accumulation occurred in cells in the decidual bed at both the mesometrial and antimesometrial poles. The embryo, especially the trophoblast giant cells, also accumulated VEGF mRNA on day 8. The expression of the VEGF receptors, Flk-1 and Flt-1, was also examined. A single transcript (approximately 6.5-7.0 kb) for Flk-1 mRNA and two transcripts (approximately 6.5 and 7.5 kb) for that of Flt-1 were detected in poly(A)+ uterine RNA samples. In situ hybridization studies showed accumulation of Flk-1 mRNA in a subset of cells in the stromal bed on day 4, but not in any uterine cell types on day 1. On days 5-8, cells in both the mesometrial and antimesometrial decidual beds exhibited accumulation of Flk-1 and Flt-1 mRNAs. Lectin binding (Dolichos biflorus agglutinin) was used to identify newly sprouting endothelial cells (angiogenesis), while an antibody to the von Willebrand factor (vWF) was employed to identify endothelial cells in general.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 7490566 TI - Changes in lipid peroxidation and free radical scavengers in the brain of hyper- and hypothyroid aged rats. AB - To determine how lipid peroxides and free radical scavengers are changed in the brain of hyper- or hypothyroid rats, we examined the behavior of lipid peroxide and free radical scavengers in the cerebral cortex of aged (1.5 years old) rats that had been made hyper- or hypothyroid by the administration of thyroxine or methimazol for 4 weeks. Concentrations of catalase, Mn-superoxide dismutase (SOD) and glutathione peroxidase (GSH-PX) were increased in hyperthyroid rats compared with euthyroid rats. Concentrations of total SOD, Cu,Zn-SOD and GSH-PX were increased but that of Mn-SOD was decreased in hypothyroid animals. There were no differences among hyperthyroid, hypothyroid and euthyroid rats in the levels of coenzymes 9 or 10. The concentration of lipid peroxides, determined indirectly by the measurement of thiobarbituric acid reactants, was decreased in hyperthyroid rats but not in hypothyroid rats when compared with euthyroid animals. These findings suggest that free radicals and lipid peroxides are scavenged to compensate for the changes induced by hyper- or hypothyroidism. PMID- 7490567 TI - The relative importance of the oligosaccharide units in human chorionic gonadotropin (CG) for LH/CG receptor activation in rat Leydig cells and mouse Leydig tumor cells. AB - The biological properties of deglycosylated human chorionic gonadotropin (DhCG), obtained by hydrogen fluoride treatment (HF-DhCG) of intact hCG or by oligonucleotide-directed mutagenesis (CHO-DhCG), and that of their fully glycosylated counterparts, were tested in terms of cAMP and steroid production in rat Leydig cells and in mouse Leydig tumor cells (MA-10 cells). In both cell types, HF-DhCG and CHO-DhCG possessed comparable biological activities. The maximum for DhCG-induced cAMP production was approximately 12% of that of intact hCG when tested in rat Leydig cells, and only 2% when tested in MA-10 cells. DhCG possessed significant steroidogenic activity in both cell types. In MA-10 cells the maximum for DhCG-induced steroidogenesis was 30-50% of that of intact hCG, while in rat Leydig cells DhCG and hCG induced similar steroidogenic maxima. Based on its ED50, DhCG possessed 10-17% of the steroidogenic potency of intact hCG in rat Leydig cells, while in MA-10 cells DhCG was only 2-fold less potent than hCG. When accurate hormone-receptor binding data are absent, the intrinsic receptor-stimulating activity of a ligand can still be estimated at full receptor occupancy, provided that over the whole dose range the biological response is proportional to receptor stimulation. The present data show that in transfected MA-10(P+29) cells which over-express rat phosphodiesterase, the hormone-induced stimulation of cAMP and steroid production is directly coupled to receptor activation up to maximal occupation of the LH/CG receptor.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7490568 TI - Enhanced hypo-osmoregulation induced by warm-acclimation in antarctic fish is mediated by increased gill and kidney Na+/K(+)-ATPase activities. AB - Serum osmolality and serum inorganic ion concentrations were studied in two antarctic fish species, Trematomus bernacchii and T. newnesi, during 5 weeks of acclimation to 4 degrees C and compared with control values for groups acclimated to -1.5 degrees C. Acclimation to 4 degrees C significantly decreased the serum osmolality of both species, thereby increasing their seawater-to-extracellular fluid (ECF) osmotic gradient. The decline in osmolality with acclimation to 4 degrees C was accompanied by significant and rapid losses of Na+ and Cl- during the first 14 days of acclimation and was maintained throughout the study period. At day 35 of acclimation, the lipid composition and microsomal Na+/K(+)-ATPase specific activities at 4 degrees C and 37 degrees C were determined in membranes from gill, kidney, liver and muscle tissues. No warm-induced decrease in fatty acid unsaturation was found in the tissues of either species. In the gills and kidneys of both species, the Na+/K(+)-ATPase activities assayed at 4 degrees C were increased after acclimation to 4 degrees C. The Na+/K(+)-ATPase activities at 37 degrees C increased at the higher acclimation temperature in T. newnesi kidneys and T. bernacchii gills, but in both species there was no compensation to temperature in the liver, regardless of assay temperature. Muscle Na+/K(+)-ATPase activity decreased in response to warm-acclimation in T. bernacchii and T. newnesi assayed at 4 degrees C and 37 degrees C, respectively. During acclimation to 4 degrees C, the discontinuity in the Arrhenius plot of the Na+/K(+)-ATPase activities of T. newnesi gill moved to a lower temperature, whereas that of kidney remained unchanged. The results indicate that acclimation to 4 degrees C induced a decrease in serum osmolality which resulted from the positive compensation of Na+/K(+)-ATPase in osmoregulatory tissues. The enhancement in Na+/K(+)-ATPase activity at 4 degrees C suggests that energy expenditure in antarctic fish may be lessened, in part, by maintaining a reduced seawater-to-ECF osmotic gradient. PMID- 7490569 TI - Effect of pharmacological blockade on cardiovascular responses to voluntary and forced diving in muskrats. AB - Neural control of free and forced diving bradycardia and peripheral resistance was studied in the muskrat (Ondatra zibethicus) by means of acute pharmacological blockade with the muscarinic blocker atropine, the alpha-adrenergic blocker phentolamine and the beta-adrenergic blockers nadolol and propranolol. Saline injection was used as a control. Heart rate in control animals increased before voluntary dives and dropped markedly as soon as the animals submerged. Heart rate started increasing towards the end of voluntary dives and reached pre-dive values within the first 5 s of recovery. Pre-dive and post-dive tachycardia were reduced in beta-blocked animals, emphasizing the role of the sympathetic system during the preparatory and recovery periods of voluntary dives. Diving bradycardia and the acceleration in heart rate before surfacing were abolished by atropine and unaffected by nadolol, demonstrating the importance of vagal efferent activity during diving. The results after blockade with nadolol suggest that there is an accentuated antagonism between the two branches of the autonomic nervous system during diving, so that parasympathetic influences on the heart predominate. Propranolol-treated muskrats had a higher diving heart rate than saline- and nadolol-treated animals, which may be due to a sedative effect caused by propranolol crossing the blood-brain barrier, a blockade of central catecholaminergic pathways or a peripheral neural effect, due to the anaesthetic properties of propranolol. Phentolamine did not affect diving bradycardia, indicating that diving bradycardia occurs independently of peripheral vasoconstriction.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7490570 TI - Sensitivity of the mystacial vibrissae of harbour seals (Phoca vitulina) for size differences of actively touched objects. AB - We studied the capability of one male and one female harbour seal Phoca vitulina to discriminate diameter differences of circular discs by means of active touch with their mystacial vibrissae. To prevent the animals from perceiving visual information, they were blindfolded during trials. In a two-alternative forced choice procedure, the seals were required to choose the larger of two Perspex discs. Weber fractions c (the ratio of the lowest diameter difference detected by the seals on 75% of occasions to the starting disc diameter D, delta D/D = c) were determined for three standard discs (diameters 1.12cm, 5.04cm and 8.74cm) by the psychophysical method of limits. While the seals achieved Weber fractions of 0.29 (male) and 0.26 (female) at the smallest standard disc, their performance improved with increasing disc size, resulting in an approximately constant Weber fraction of 0.13 (male) and 0.08 (female) for the two larger standard discs. The difference in performance between the two seals did not reflect a real difference in sensitivity, but may best be explained by a difference in choice behaviour. As a measure of tactile acuity, the Weber fractions obtained for the larger standard discs indicate that harbour seals can use their mystacial vibrissae as efficiently for active touch as monkeys use their hands. PMID- 7490571 TI - Non-linear dynamics of neurochemical modulation of mosquito oviduct and hindgut contractions. AB - Microphysiological techniques and computer-based methods of data acquisition and analysis were used to investigate the dynamics of neurochemical action on spontaneous contractions of continuously perfused oviducts and hindguts isolated from female mosquitoes (Aedes aegypti). Proctolin, leucomyosuppressin (LMS), serotonin and octopamine, which are known to modulate insect muscle contractions, and a mosquito neuropeptide, Aedes Head Peptide I (Aea-HP-I), which inhibits host seeking behavior, were tested in the bioassays. LMS depressed contractile activity in oviducts at concentrations above 10(-12) mol l-1, but hindguts did not respond to concentrations below 10(-6) mol l-1. Hindgut contractions spontaneously restarted in 10(-6) mol l-1 LMS, but only washing out LMS restored activity in oviducts. LMS changed the amplitude of the oviduct contractions, but the dynamics of contraction remained steady. Following recovery of contractions in LMS, hindgut tissues contracted with a more regular pattern. Serotonin and octopamine had an identical action on oviduct and hindgut tissues. At concentrations greater than 10(-8) mol l-1, serotonin eliminated the refractory period between contractions and thus increased the contraction frequency of oviducts. Though the contractions appeared chaotic, state-series analysis proved that serotonin-treated oviducts contracted rhythmically. Above 10(-6) mol l-1, serotonin extinguished normal hindgut peristalsis and induced regular fluttering of the anteriormost portion of the hindgut, an effect possibly related to diuresis following the blood meal. Proctolin failed to stimulate both oviduct and hindgut contractions at concentrations up to 10(-6) mol l-1, but at 10(-8) mol l 1 induced contractions of cricket hindgut preparations. Aea-HP-I had no effect on either tissue at 10(-6) mol l-1 or lower concentrations. State-series analysis, based on simple manipulations of experimental data, permitted direct observation of the dynamics of oviduct and hindgut contractile activity. PMID- 7490572 TI - Characterization and modulation of Na+ and Ca2+ currents underlying the action potential in bag cells of two species of Aplysia. AB - The neurosecretory bag cells of Aplysia produce long trains of action potentials (afterdischarge) to release hormones important to egg laying. These ionic currents are modulated by second messengers. Modulation of excitability in bag cells is incompletely understood partly because the currents that are modulated have not been fully characterized. Whole-cell voltage-clamp experiments were executed in cultured cells from sexually mature A. californica and A. brasiliana to characterize the inward voltage-gated currents for Na+ and Ca2+. These species had similar Na+ and Ca2+ current characteristics. The Na+ currents activated at voltages less negative than -30 mV and peaked at between +10 and +20 mV in artificial sea water. The time course and pharmacology of bag cell Na+ currents were similar to those of fast Na+ current in other excitable cells. Na+ currents were abolished in Na(+)-free extracellular solution and were not inhibited by Cd2+. The KD for inhibition by tetrodotoxin was 2.6 nmol l-1. The Na+ current was relatively insensitive to depolarized holding potentials (Vh), maintaining approximately 65% of peak current amplitude throughout the activation range at Vh = -30 mV. In experiments using a 1 s depolarized Vh prior to a test pulse, the half-inactivation voltage (V1/2) was -21 mV. The time constant of recovery from steady-state activation was 2.9 ms at Vh = -70 mV and 6.8 ms at Vh = -30 mV. The Ca2+ currents activated near -10 mV and peaked at approximately +20 mV with 11 mmol l-1 Ba2+ as the charge carrier. The pharmacology and V1/2 of bag cell Ca2+ current were similar to those of L-type Ca2+ currents. In extracellular solution without Na+, but containing Ba2+, Cs+ and tetraethylammonium, the Ca2+ current was inhibited by 25-100% by nifedipine (10 mumol l-1), mean 42%, and was unaffected in the majority of cells by omega-conotoxin (10 mumol l-1). The Ca2+ current was insensitive to Ni2+ (100 mumol l-1), but was abolished by 100 mumol l 1 Cd2+. Like the Na+ current, the Ca2+ current was relatively insensitive to depolarized Vh, maintaining more than 80% of peak current amplitude throughout the activation range at Vh = -40 mV. With a 1 s depolarized Vh prior to a test pulse, the V1/2 was -30 mV.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 7490573 TI - The structure, response properties and development of a hair plate on the mesothoracic leg of the locust. AB - A hair plate is present on the proximal anterior face of the pro- and mesothoracic tibiae of the legs of the locust Schistocerca gregaria, but not on the metathoracic legs. The hair plate is in a depression of the cuticle and contains about 11 hairs, which are all polarised with their tips pointing towards the dorsal surface of the tibia. The hairs are all of the same trichoid sensilla type and vary in length from 90 to 140 microns. Associated with the hair plate is a pronounced distal extension of the anterior femoral coverplate, the inner face of which is concave, that makes contact with the hairs during flexion and extension movements of the tibia. During postembryonic development, no tibial hair plate hairs are present in the first four larval stages. In fifth-instar larvae just three hairs are present, while the full complement is attained only after the final moult to adulthood. The distal extension of the posterior coverplate is present through all instar stages, becoming more pronounced after each moult. Sensory neurones innervating the hairs of an adult may be divided into two classes on the basis of their responses. The first type responds phasically to imposed deflections and is velocity-sensitive. The second type responds phasotonically and is also sensitive to the velocity of the stimulus but has an additional tonic component sensitive to maintained angular deflections. Both types of afferents are directionally sensitive and respond best to deflections against the natural bend of the hair, equivalent to extension movements of the tibia.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7490574 TI - Automaticity and word perception: evidence from Stroop and Stroop dilution effects. AB - The Stroop effect is cut in half by adding a neutral word to the display. D. Kahneman and D. Chajczyk's (1983) "attention capture" account of "Stroop dilution" holds word recognition to be involuntary but strictly serial. The authors compared attention capture to 3 alternatives involving parallel rather than serial processing: In the lexicon, activation is divided among multiple words; postlexically, multiple words race for access to response processes; or prelexically, feature processing is degraded by multiple patterns whether or not they are words. Results support the latter. Multiple patterns are processed in parallel. If any are color words, Stroop effects occur but are reduced because any color word's input to lexical memory is lower in quality than if a single color word were the only pattern. Thus, lexical encoding is involuntary but can operate on several input representations in parallel, with effectiveness determined by input quality. PMID- 7490575 TI - Eye fixation patterns among dyslexic and normal readers: effects of word length and word frequency. AB - Eye fixation patterns of 21 dyslexic and 21 younger, nondyslexic readers were compared when they read aloud 2 texts. The study examined whether word-frequency and word-length effects previously found for skilled adult readers would generalize equally to younger dyslexic and nondyslexic readers. Significantly longer gaze durations and reinspection times were found for low-frequency and long words than for high-frequency and short words. The effects also showed up in the number of fixations on the target words. The effects did not differ significantly for the 2 experimental groups. The results run counter to the oculomotor dysfunction hypothesis of dyslexia. Instead, they support the view that both dyslexic and nondyslexic readers' eye fixation patterns reflect their difficulties in successfully identifying words in a text. PMID- 7490577 TI - On-line evidence for elaborative logical inferences in text. AB - A model of propositional-logic reasoning proposed by M. D. S. Braine, B. J. Reiser, and B. Rumain (1984) claims that inferences such as "p or q; not p/therefore q" are made spontaneously by readers at the moment both premises are available. This claim is inconsistent with some evidence in the text-processing literature that suggests that only those inferences necessary for textual coherence are made spontaneously. In the present study, participants read stories in which a logical inference was not necessary to maintain textual coherence, and inference making was assessed with on-line probes. Two experiments tested logical forms central to Braine et al.'s model, and both indicated that participants were making the logical inferences. Two further experiments replicated this result with stories that did not begin with thematic titles. These findings support Braine et al.'s prediction that some propositional-logic inferences are made routinely in texts that do not require them for coherence. PMID- 7490576 TI - Role of context in accessing distant information during reading. AB - In 3 experiments, the authors investigated factors that influence accessibility of backgrounded goal information. Participants read texts consisting of 2 episodes. In the 1st episode, the goal was satisfied or unsatisfied. Following a statement of goal satisfaction or goal postponement, there was a 2nd unrelated episode. After completion of the 2nd episode, target sentences were presented that were consistent with the 2nd episode but were inconsistent with completion of the earlier unsatisfied goal. Participants noticed the inconsistency only when the conflicting information was separated by a few sentences or when the context of the goal had been reinstated. The results are consistent with the resonance model described by J. E. Albrecht and E. J. O'Brien (1993) and by J. L. Myers, E. J. O'Brien, J. E. Albrecht, and R. A. Mason (1994). PMID- 7490578 TI - Causal inferences in reading: from immediate activation to long-term memory. AB - Three different tasks were used to investigate the time course of drawing causal inferences. Participants read passages that contained a causal coherence break that could be resolved by reactivating a concept presented earlier in the passage. In Experiment 1, participants named a probe word that represented the earlier mentioned cause more quickly after encountering the causal coherence break, suggesting that the causal concept had quickly been reactivated. In Experiment 2, participants were slow to read a sentence after the causal coherence break that contradicted the intended inference, indicating that the inference had been encoded and retained in working memory. In Experiment 3, the results of a recall task indicated that the causal link was also included in the long-term memory representation of the text. PMID- 7490579 TI - Perceived necessity explains the dissociation between logic and meaning: the case of "only if". AB - The purpose of this article was to investigate why sentences of the form "p only if q" often appear to have a different meaning than sentences of the form "if p, then q," despite their logical equivalence. The results of 3 experiments indicate that when "if ... then" statements were equated with respect to necessity and temporal relations, different pragmatic relations (such as permission, causation, etc.) elicited similar "only if" judgments. However, different necessity relations elicited different "only if" judgments, regardless of the type of pragmatic relation expressed in the "if then" statement. These data suggest that "only if" judgements are primarily mediated by necessity and temporal relations and that pragmatic contexts may play a more indirect role, such as in the interpretation of necessity and temporal relations. Suggestions for how these findings might be incorporated into pragmatic schema theory (P. W. Cheng & K. J. Holyoak, 1985; P. W. Cheng, K. J. Holyoak, R. E. Nisbett, & L. M. Oliver, 1986) and mental models theory (P. N. Johnson-Laird & R. M. J. Byrne, 1991) are discussed. PMID- 7490580 TI - Perceptual and nonperceptual components of implicit memory for pictures. AB - The transfer appropriate processing (TAP) framework posits that in data-driven tasks, such as picture naming (PN) or picture perceptual identification, repetition priming is greater when perceptual processes engaged at study are recapitulated at test. Thus, priming with pictures is greater after study-phase exposure to pictures than to words (picture names). A. S. Brown, D. R. Neblett, T. C. Jones, and D. B. Mitchell (1991) reported that a pure-list format eliminated perceptual priming: Participants who saw either pictures or words in a study phase showed equal priming in a PN task. In the present study, participants showed greater priming after exposure to pure lists of pictures than to pure lists of words in 3 PN and 1 picture perceptual identification experiments. Thus, perceptual priming occurred in 4 pure-list picture priming tasks, as predicted by the TAP framework. Priming also was found after exposure to words. In PN and picture perceptual identification tasks, implicit memory for pictures includes perceptual and nonperceptual components. PMID- 7490581 TI - Verbal vulnerability of perceptual expertise. AB - Three experiments explored the role of perceptual expertise in mediating the finding (termed verbal overshadowing) that describing a face can impair later recognition. In Experiment 1, verbalization impaired White participants' recognition of White faces (expert domain) but not African American faces (novice domain). In Experiment 2, judges attempted to identify targets on the basis of the verbal descriptions generated in Experiment 1. Experiment 2 revealed a significant relationship between verbalization participants' recognition performance and yoked judges' identification performance for other-race but not own-race faces, suggesting that other-race recognition may involve a unique reliance on "verbalizable" information. In Experiment 3, the interaction between verbalization and race of face was replicated with upright faces but was attenuated with inverted recognition arrays (a manipulation that reduces the influence of configural information). Collectively, these findings suggest that verbalization may disrupt the nonreportable configural processes associated with recognizing stimuli with which one is an expert. PMID- 7490582 TI - Types and tokens unscathed: a reply to Whittlesea, Dorken, and Podrouzek (1995) and Whittlesea and Podrouzek (1995). AB - N. G. Kanwisher (1987; J. Park & N. G. Kanwisher, 1994) has explained repetition blindness in terms of a distinction in visual perception between type activation and token individuation; repeated items are successfully recognized (matched to stored types) but are less likely than unrepeated items to become individuated as separate perceptual tokens. Whittlesea and colleagues (B. W. A. Whittlesea, M. D. Dorken, & K. W. Podrouzek, 1995; B. W. A. Whittlesea & K. W. Podrouzek, 1995) argued that repetition blindness does not reflect different processing of repeated and unrepeated items but is better explained as the result of a combination of separate but nondistinctive processing of repeated items and postlist report biases. However, we argue that none of the results reported by Whittlesea and colleagues are inconsistent with the token-individuation hypothesis. PMID- 7490583 TI - "Where did I go wrong?" A psychophysiological analysis of error detection. AB - There is a component of the event-related brain potential, the error-related negativity (or ERN), that is related to error detection in choice reaction time tasks. The J. Miller (1982) paradigm was used to determine whether the detection process manifested by the ERN involves a comparison between representations of the actual response and the correct response or between representations of the stimulus anticipated by the subject and the stimulus that actually occurs. The data favored the former rather than the latter kind of comparison, with the magnitude of the error signal depending on the similarity or dissimilarity between the two response representations. In turn, response similarity depended on the strategy used by the subjects to select responses: Response parameters selected first defined which responses would be most similar. PMID- 7490584 TI - Inhibition of return to successively cued spatial locations. AB - Inhibition of return refers to a bias against returning attention to a location that has been recently attended. Experiments are reported that examined inhibition of return to multiple exogenously cued spatial locations. When 2 peripheral locations were cued in succession, inhibition was found for only the 1 most recently cued location. In addition, more inhibition occurred at the location of the most recent cue if the earlier cue had also been presented there, as compared with an earlier cue at a different location. Thus, the magnitude of the inhibition for a location appears to depend on the effectiveness of the attentional cue to that location. Other results suggest that candidate locations for inhibition are displaced by subsequent cues--they do not simply decay. The results provide an initial framework within which to study inhibition of return to multiple spatial locations. PMID- 7490586 TI - Allocating visual attention: tests of a two-process model. AB - Observers require less time to identify a visual target when its location is cued in advance than when it is not cued, and the magnitude of the improvement depends on the validity of the cue. According to J. Jonides's (1983) 2-process model, there exist 2 possible modes of attentional readiness: a focused-attention mode and a diffuse-attention mode. Observers are assumed to enter the focused attention mode on a proportion of trials that matches the validity of the cue and to enter the diffuse-attention mode on the remaining trials. The present experiment tested and rejected the response time mixture prediction of the 2 process model. An instance of the class of 1-process models in which perceptual objects are sampled in parallel according to the validity of the cue was evaluated. A stochastic simulation of the model yielded results that paralleled those of the experiment. PMID- 7490585 TI - Loosening the constraints on illusory conjunctions: assessing the roles of exposure duration and attention. AB - Illusory conjunctions are the incorrect combination of correctly perceived features, such as color and shape. They have been found to occur using a brief exposure (under 200 ms) and a dual task designed to divert attention. The present study investigated the roles of exposure duration and attention in obtaining illusory conjunctions. Several mathematical models of the feature integration task were also assessed. Experiment 1 tested participants' accuracy at combining features using a long exposure and an attention-diverting taks. Experiment 2 compared performance with and without the attention-diverting task. The final experiment compared performance using a brief (0.15 s) and a long (1.5 s) exposure duration without an attention-diverting task. Neither attention nor exposure duration had a significant effect on feature integration. PMID- 7490587 TI - Constraints on the processing of rolling motion: the curtate cycloid illusion. AB - When a wheel rolls along a flat surface, a point on its perimeter traces a cycloid trajectory, forming a sequence of adjacent semicircle-like scallops. However, when mentally visualizing this point's trajectory, participants erroneously describe the point's path as looping back on itself between each scallop or phase of the cycloid, a phenomenon called the curtate cycloid illusion. The studies supported the hypothesis that the curtate cycloid illusion occurs because the cognitive system sometimes does not have sufficient resources for simultaneously processing 2 components of the motion: its translation and its rotation about its current instant center. Four experiments using computer animated rolling wheels found that participants who were high in spatial ability were less susceptible to the curtate cycloid illusion than were low-spatial participants, that high-spatial participants were not susceptible to the illusion if they could control the animated wheel display, and that the illusion was substantially decreased if the opportunity to compute instant centers was reduced. PMID- 7490589 TI - Dynamics and the orientation of kinematic forms in visual event recognition. AB - The authors investigated event dynamics as a determinant of the perceptual significance of forms of motion. Patch-light displays were recorded for 9 simple events selected to represent rigid-body dynamics, biodynamics, hydrodynamics, and aerodynamics. Observers described events in a free-response task or by circling properties on a list. Cluster analyses performed on descriptor frequencies reflected the dynamics. Observers discriminated hydro- versus aerodynamic events and animate versus inanimate events. The latter result was confirmed by using a forced-choice task. Dynamical models of the events led us to consider energy flows as a determinant of kinematic properties that allowed animacy to be distinguished. Orientation was manipulated in 3 viewing conditions. Descriptions varied with absolute display orientation rather than the relative orientation of display and observer. PMID- 7490588 TI - Cross-modal discrepancies in coarticulation and the integration of speech information: the McGurk effect with mismatched vowels. AB - Two experiments examined the impact of a discrepancy in vowel quality between the auditory and visual modalities on the perception of a syllable-initial consonant. One experiment examined the effect of such a discrepancy on the McGurk effect by cross-dubbing auditory /bi/ tokens onto visual /ga/ articulations (and vice versa). A discrepancy in vowel category significantly reduced the magnitude of the McGurk effect and changed the pattern of responses. A 2nd experiment investigated the effect of such a discrepancy on the speeded classification of the initial consonant. Mean reaction times to classify the tokens increased when the vowel information was discrepant between the 2 modalities but not when the vowel information was consistent. These experiments indicate that the perceptual system is sensitive to cross-modal discrepancies in the coarticulatory information between a consonant and its following vowel during phonetic perception. PMID- 7490590 TI - Is human object recognition better described by geon structural descriptions or by multiple views? Comment on Biederman and Gerhardstein (1993). AB - Is human object recognition viewpoint dependent or viewpoint invariant under "everyday" conditions? I. Biederman and P.C. Gerhardstein (1993) argued that viewpoint-invariant mechanisms are used almost exclusively. However, our analysis indicates that (a) their conditions for immediate viewpoint invariance lack the generality to characterize a wide range of recognition phenomena, (b) the extensive body of viewpoint-dependent results cannot be dismissed as processing "by-products" or "experimental artifacts," and (c) geon structural descriptions cannot coherently account for category recognition, the domain they are intended to explain. The weight of current evidence supports an exemplar-based multiple views mechanism as an important component of both exemplar-specific and categorical recognition. PMID- 7490591 TI - Epidemiology of aortic aneurysm in the United States. AB - Recent apparent increases in occurrence of aortic aneurysm were reported for abdominal aortic aneurysm from several countries. In order to assess U.S. trends, an analysis of mortality and hospitalization data from the National Center for Health Statistics for aortic aneurysm in the United States in 1979-1992 was performed. In 1991, 16,696 deaths were attributed to aortic aneurysm, abdominal aneurysm accounting for 52%. Between 1979 and 1990, dissecting aneurysm death rates showed inconsistent changes in males and slight increases in females. Age adjusted rates were higher in blacks than whites, and in males than females. Death rates for abdominal aneurysms showed slight decreases in white males and slight increases in black males and white females. Rates were higher in whites than blacks, and in males than females. The number of hospital discharges with a first-listed diagnosis of aortic aneurysm increased from 39,000 in 1979 to 67,000 in 1992. The rate of diagnoses increased from 1979 to 1984 with no consistent change thereafter for total and abdominal aneurysms, which comprised over 75% of total diagnoses. The number of all-listed aortic aneurysm resections with graft replacement increased from 10,000 in 1979 to 40,000 in 1988 with no consistent change thereafter. Increased utilization of diagnostic ultrasound of the abdomen and retroperitoneum leading to improved case finding for abdominal aneurysms may have been one cause of increasing hospital discharge rates prior to 1985. Continued monitoring of national data on mortality and morbidity from aortic aneurysms is desirable to assess effects of diagnostic, therapeutic, and preventive interventions. PMID- 7490592 TI - Hierarchical structure in ADL and IADL: analytical assumptions and applications for clinicians and researchers. AB - The results of a Canadian study have shown that a set of 12 (I)ADL items did not meet the criteria of Guttman's scalogram program, questioning the assumption of hierarchical ordering. In this article, the hierarchical structure of (I)ADL items from the Canadian elderly sample is retested with another scalogram program, and both the epidemiological research and clinical applications of hierarchical scales are discussed. For retesting the hierarchical structure, Mokken's Stochastic Scale Program was used. When item distributions were not too skewed, the existence of one general dimension with two (correlated) subdimensions were clearly found. The analyses for scalability showed that, except in the case of extremely skewed item distributions, the data set met the criteria of Mokken's Stochastic Scale Program, indicating a hierarchical structure of the items. The scalability of a set of items is affected by the sample studied, the choice of response options for each item, the selection and number of items in the scale, and the type of scaling procedure. The practical assumptions of hierarchy should be restricted to program planning and epidemiological research purposes only. For clinical assessment, a finer grading is desirable and all questions on an ADL/IADL inventory should be asked for each individual. PMID- 7490593 TI - Results and efficacy of public screening for hypercholesterolemia: the Bavarian Cholesterol Screening Project. AB - Hypercholesterolemia is a risk factor for the development of coronary disease. It does not present with symptoms and can be diagnosed by laboratory examination only. Screening is the only means to detect subjects at risk at a time when preventive measures can effectively be applied. We therefore initiated the Bavarian Cholesterol Screening Project (BCSP). Occasional screening was performed in 150,089 subjects (81,286 women, 68,803 men) in 214 campaigns. The mean cholesterol value was 243 +/- 52 mg/dl for women and 231 +/- 53 mg/dl for men; 37.3% of women and 38.1% of men had values of 201-250 mg/dl, 42.2% of women and 33.7% of men values > 250 mg/dl. Also, the following risk factors were recorded: smoking in 11.3% of women and in 20.2% of men, hypertension in 19.8% of women and 17.4% of men, diabetes mellitus in 4.2% of women and 4.1% of men, obesity in 16.8% of women and 20.9% of men, and a family history of myocardial infarction in 34.8% of women and 26.0% of men. Of the 27,084 men who had their cholesterol checked for the first time, 35.6% had levels between 201 and 250 mg/dl, and 22.9% had levels above 250 mg/dl. Of the 27,870 women whose cholesterol level had not been checked before, 38.8% had levels between 201 and 250 mg/dl, and 27.1% had levels above 250 mg/dl. More than 70% of the subjects with levels between 200 and 250 mg/dl had at least one additional risk factor. We conclude, on the basis of this study, that the risk factor hypercholesterolemia is unknown in as much as 20% of the population of Bavaria. These newly detected subjects were offered the opportunity to recognize this risk and take subsequent measures of prevention. Screening projects for hypercholesterolemia therefore can be an effective means of improving public health. PMID- 7490594 TI - Varicella zoster virus and multiple sclerosis in a Hutterite population. AB - There are similarities between multiple sclerosis and varicella. They are common in the same parts of the world and both are scarce in other areas. Immigration studies suggest the environmental cause of multiple sclerosis (MS) must be contracted prior to age 15 years and will usually remain dormant for years. At age 10 years varicella has occurred in greater than 95% of children living in the high-risk areas for both of these diseases. The varicella zoster virus (VZV) could be etiologically important in multiple sclerosis. The known host containment of the virus for decades with recrudescence and the variable cell mediated immunity of the host, which can wax and wane without clinical manifestations, all lend themselves to the natural history of multiple sclerosis. A population-based study of the medical records of 5601 Hutterite Brethren was performed to determine the occurrence of multiple sclerosis, varicella, and herpes zoster. Compared to their matched non-Hutterite neighbors who acted as controls, these events were significantly less common among the Hutterites. Included in the study was an assessment of other common neurological diseases and "autoimmune" diseases among the Hutterites and the controls. There is evidence of a relationship between MS and VZV that may not be coincidental. PMID- 7490596 TI - An addition to the controversy on sunlight exposure and melanoma risk: a meta analytical approach. AB - Case control studies on the association between sunlight exposure and melanoma risk show considerable differences in design; this could be responsible for the variation in study results. In an attempt to resolve the controversy between study results, the results of 25 publications on case control studies were evaluated using meta-analytical techniques. Comparison of odds ratios between subgroups of studies revealed that the range of odds ratios was far greater for hospital-based studies than for population-based studies. For the latter type of studies, the odds ratios were homogeneous and the pooled odds ratios were 1.57 (95% confidence interval [CI], 1.29-1.91) for intermittent sunlight exposure and 0.73 (95% CI, 0.60-0.89) for chronic exposure. However, among other problems, the lack of standardized measures for sunlight exposure warrants cautious interpretation of these results. It is concluded that evidence to support the intermittent sunlight theory is still far from complete. PMID- 7490595 TI - Timing payments to subjects of mail surveys: cost-effectiveness and bias. AB - Although mailed surveys are an important component of epidemiological research, results from mailed surveys are often suspect because of poor response rates and the potential for nonresponse bias. Previous work has demonstrated that paying subjects to complete questionnaires increases response rates, but this work has not well addressed the impact of the timing of incentives on total cost, cost effectiveness, and response bias. We surveyed 400 university employees about health benefits. By random allocation, half received a check for $5 along with the mailed survey, and the other half received the promise of $5 on return of a completed survey. The response rates for both groups were about the same (64 and 59%, respectively), but prepayment was less expensive in aggregate and less expensive per response. In addition, we found that subjects with lower salaries were more likely to respond when paid in advance. We conclude that prepayment may actually be less expensive and more cost effective than payment on completion, but that the timing of payment may influence the profile of respondents. PMID- 7490597 TI - Cerebrovascular disease in children under 16 years of age in the city of Dijon, France: a study of incidence and clinical features from 1985 to 1993. AB - Our motivation for undertaking the present survey was to determine the incidence, the distribution, and the clinical features of ischemic and hemorrhagic strokes in children under 16 years old, in a well-defined population-based study. The survey was carried out on the population of the City of Dijon (150,000 inhibitants) from January 1, 1985 to December 31, 1993, collecting prospectively both in adulthood and in childhood (23,877 resident children). Diagnosis of stroke was established on the basis of clinical features and the mechanism was identified by CT scan from 1985 to 1987, and by CT scan and magnetic resonance imaging from 1987 to 1993. When a hemorrhagic stroke was identified, a cerebral arteriogram and an investigation of the coagulation factors were performed. When an ischemic stroke was identified, the following were performed: an ultrasound examination of the cervical arteries, a cerebral arteriogram, a lumbar puncture, an investigation of the coagulation factors and lipid status, a measurement of homocysteine in the plasma and the urine, an electrocardiogram (EKG), a Holter procedure, and a cardiac echography. During the 9 full calendar years of this study we observed 28 stroke patients from a population of 23,877 resident children. There were 17 cases of ischemic stroke, representing some 61% percent of the total, as well as 11 cases of hemorrhagic stroke, 39% percent of the total. The average annual incidence rate was 13.02/100,000 for all strokes, 7.91/100,000 for ischemic strokes, and 5.11/100,000 for hemorrhagic strokes.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7490598 TI - Physical-strength tests and mortality among visitors to health-promotion centers in Japan. AB - A follow-up study was conducted to clarify the relationship between physical strength level and risk of death from all causes and from cancer and cardiovascular disease. The 7286 persons who were examined at seven health promotion centers throughout Japan between 1982 and 1987 were followed up. By January 1992, 6259 persons (85.9%) had been contacted by questionnaire. They included 3117 men (49.8% of all subjects studied) (average age 53.6 years at baseline, SD = 9.0 years, range 40-84 years), and 3142 women (50.2%) (average age 54.5 years at baseline, SD = 8.5 years, range 40-85 years). The follow-up period for each person averaged 6.1 years, for a total of 38,253 person-years. During this period, 155 deaths were reported. At baseline, five physical-strength tests (grip strength, side step, vertical jump, standing trunk flexion, and sit-ups) were performed. Five clinical laboratory tests (thickness of skinfold, blood sugar, total serum cholesterol, percent vital lung capacity, and blood pressure) were also conducted. The examinees were questioned about smoking status (current smoker, nonsmoker, and ex-smoker). Men with thicker skinfold [relative risk (RR) = 2.11] and higher levels of blood sugar (RR = 1.89) had an excess risk of death from all causes. Men with higher serum cholesterol (RR = 5.08), thicker skinfold (RR = 4.54), and elevated blood pressure (RR = 2.33) had an excess risk of death from cardiovascular disease. In women, no relationship was seen between clinical laboratory tests and an excess risk of death. Men exhibiting lower values for side step (RR = 2.43), vertical jump (RR = 2.37), sit-ups (RR = 1.93) and grip strength (RR = 1.92) also had an excess risk of death from all causes. Furthermore, men with lower heights for vertical jump (RR = 5.51) had an excess risk of death from cardiovascular disease. After adjustment for skinfold thickness, blood sugar, total serum cholesterol, blood pressure, percent vital lung capacity and smoking status, men with a lower level of side step, vertical jump, and grip strength had an excess risk of death from all causes. No such relationship was seen between physical-strength level and an excess risk of death in women. It is concluded that a low level of physical strength might be significantly correlated with subsequent health outcomes in men. PMID- 7490599 TI - Reliability of near-infrared interactance body fat assessment relative to standard anthropometric techniques. AB - We examined the repeatability of near-infrared interactance (NIR) body fat determination as compared with that of body mass index (BMI), waist-to-hip ratio (WHR), and waist girth. Thirty-nine volunteers (16 men, 23 women) had percent body fat (%BF) measurements made with a portable NIR device as well as the standard anthropometric indices of height, weight, waist girth, and hip circumference. Frame size and physical activity levels were also determined. For each participant, three independent measurements of each index were made by two trained readers during a 2-week period. The two readers varied significantly in their measurement of %BF and hip circumference. The variability in %BF was largely due to differences between the first and the second measurements, and only for one of the readers. Second and third measurements were not statistically significantly different for either reader, suggestive of a training effect. Variance component calculations revealed that the reliability of NIR is 95.3%, compared with 99.9% for BMI; 93.4% for waist girth; and 82.4% for WHR, with the majority of the remaining variance accounted for by the method itself. We conclude that the NIR method has good repeatability, with low intra- and interobserver variability, provided that readers are carefully trained. However, the NIR device offers little advantage in reliability over conventional measures of adiposity such as waist girth or BMI, and requires additional input of weight, height, frame size, physical activity level, age, and gender data to calculate %BF. Associations of NIR and other anthropometric indices with cardiovascular risk factors in this population will provide additional insight into the merit of NIR body fat assessment. PMID- 7490600 TI - Relative responsiveness of condition-specific and generic health status measures in degenerative lumbar spinal stenosis. AB - The objective of this study was to compare the relative responsiveness of a condition-specific spinal stenosis measure and two generic health status measures for outcome assessment of surgery for degenerative lumbar spinal stenosis, and to examine whether responsiveness statistics and measures of the ability to distinguish clinically important improvement rank the instruments consistently. Physical function and symptom severity scales of the spinal stenosis measure were compared to the Sickness Impact Profile (SIP) and the Roland scale, which is derived from the SIP. Responsiveness was calculated with the standardized response mean, the effect size, and Guyatt's responsiveness statistic. The discriminative ability of the instruments to distinguish patients who improved from those who did not was assessed using satisfaction with surgery as an external criterion. Minimal clinically relevant improvement was estimated using patient satisfaction as the external criterion. All responsiveness statistics revealed the same order of responsiveness; the physical function scale (SRM = 1.07) and symptom severity scales (SRM = 0.96) were more responsive than the Roland scale (SRM = 0.77) which was only slightly more responsive than the SIP (SRM = 0.69). Strikingly, the physical dimension of the SIP (SRM = 0.62) was even less responsive than the global SIP. The shape of and the area under the ROC curves showed that the physical function and symptom severity scales discriminate better between satisfied and unsatisfied patients than the Roland scale and SIP. The sensitivity to detect clinically important changes was somewhat lower at the ends of the scales, especially for the SIP and the Roland scale. Statistical approaches that assess the ability to distinguish clinically important changes and overall responsiveness statistics ranked the measures consistently. On the basis of these findings, we suggest that a condition-specific spinal stenosis measure is preferable as the primary end point in evaluative studies of degenerative lumbar spinal stenosis. PMID- 7490601 TI - Validation of a meta-analysis: the effects of fish oil in rheumatoid arthritis. AB - The purpose of this study was to validate the results of a meta-analysis showing the efficacy of fish oil in rheumatoid arthritis with the results of a re analysis of the complete primary data set. A Medline search yielded seven published papers. Three additional trials were found by contacting authorities in the field. Inclusion criteria included (1) a double-blind, placebo-controlled study, (2) use of at least one of seven predetermined outcome measures, (3) results reported for both placebo and treatment groups at baseline and follow-up, (4) randomization, and (5) parallel or cross-over design. Papers were scored for quality. Demographic and outcomes variables were collected. For the re-analysis of the primary data, the same variables were abstracted for the 395 individual patients randomized. The meta-analysis demonstrated that dietary fish oil supplementation for 3 months significantly reduced tender joint count (rate difference [RD] [95% CI] = -2.9 [-3.8 to -2.1] [p = 0.001]) and morning stiffness (RD [95% CI] = -25.9 [-44.3 to -7.5] [p < 0.01]) as compared with heterogeneous dietary control oils. The re-analysis of the primary data confirmed a significant reduction in tender joint count (p = 0.001) and in morning stiffness (p < 0.02) in the parallel analysis that ignored interaction terms. The analyses that included an interaction term between site and treatment again confirmed a significant reduction in tender joint count. The results for morning stiffness were similar to the meta-analysis, but did not quite reach statistical significance (p = 0.052-0.083). The relative improvements in the other outcome variables did not reach statistical significance. Use of fish oil improved the number of tender joints and duration of morning stiffness at 3 months as analyzed by both meta- and mega-analysis. The fuller mega-analysis confirmed the results of the meta-analysis. The advantages of mega-analysis were as follows: (1) the ability to analyze the homogeneity of the patient populations, (2) the ability to make clinically sensible adjustments in the form of the comparison, and (3) the ability to examine subsets of the data. PMID- 7490602 TI - Use of an automated prescription database to identify individuals with asthma. AB - We used medication-dispensing information for 4 years (1/1/87 through 12/31/90) to examine the utilization of anti-asthma medications among 175,562 members of a large health maintenance organization. A total of 297,863 anti-asthma medications was dispensed during the study period, over one-half of which (55%) were beta agonists, followed by aminophylline preparations (23%) and inhaled corticosteroids (13%). Next, we compared the predictive value of three algorithms for identifying individuals with asthma: (1) two or more beta-agonist dispensings, (2) both a beta-agonist and an inhaled corticosteroid dispensing, and (3) five or more total anti-asthma dispensings. We performed chart reviews for 40 subjects aged 5-45 years in each of these three groups and made a clinical judgment, based on all available information in the chart, as to whether each patient had asthma. Two levels of certainty were used: "any asthma" and "definite asthma." All 120 charts reviewed presented a clinical picture consistent with asthma. However, patients identified by the algorithm that included both a beta agonist and an inhaled corticosteroid were more likely to meet our criteria for "definite" asthma and more likely to have moderate to severe asthma. These results demonstrate the feasibility of using an automated outpatient pharmacy database to identify patients with asthma. PMID- 7490603 TI - Gangliosides and Guillain-Barre syndrome. AB - Cases of Guillain-Barre syndrome (GBS) associated with parenteral use of gangliosides have been reported in several European countries. To evaluate the hypothesis of association between ganglioside exposure and occurrence of GBS, a case-control study was conducted. GBS cases discharged during 1989 from public and private hospitals in three Italian provinces were identified: 42 GBS cases and 420 controls matched on age and gender were enrolled. Data of onset of symptoms of GBS was taken from clinical records. Exposure status of subjects was ascertained through the regional computerized drug prescription monitoring system. The odds ratio of association between ganglioside use, in the 30 days prior to onset of symptoms, and GBS was 9.1 (95% confidence interval 2.8-29.4). Although there are formidable difficulties in distinguishing prodromal therapy of GBS from drug causation, the association with ganglioside therapy is strong and supportive of the hypothesis of a role of ganglioside preparations in the occurrence of GBS. PMID- 7490604 TI - Do antidepressants cause, promote, or inhibit cancers? AB - Do antidepressants cause, promote, or inhibit cancers? OBJECTIVE: To review all human and experimental studies that examined the association of antidepressants with cancer or the effect of antidepressants on neoplastic growth. METHODS: A search was conducted of MEDLINE for relevant articles published in English between 1976 and 1993. RESULTS: Four human studies and nine experimental models were found. The human studies showed a transiently statistically significant positive association between amitriptyline and liver cancer and a negative association with pancreatic cancer; and that the antidepressants amitriptyline, nortriptyline, desipramine, and phenelzine may increase risk of breast cancer. Results of the experimental studies differed depending on which antidepressants were examined and which model was used. Amitriptyline was found to promote tumour growth, fluoxetine and clomipramine were reported to be both tumour promoters and antineoplastic agents, and imipramine and citalopram both demonstrated antineoplastic properties. CONCLUSIONS: Further epidemiologic studies in humans are needed to determine which antidepressants cause, promote, or inhibit cancers. PMID- 7490605 TI - Retrospective evaluation of patient compliance with supportive periodontal treatment. AB - Four hundred sixty-two patients were retrospectively evaluated for their compliance to periodontal therapy in terms of attendance for a recommended schedule of visits, being classified as either compliant or non-compliant. Compliant patients were further divided into erratic and complete complier groups. The results indicated that there were no significant differences between the compliant and non-compliant groups with regard to the numbers of teeth lost before treatment, and dental insurance coverage. There were more women among the compliant patients. In the non-compliant group, more patients received only initial periodontal therapy, whereas in the compliant group more patients received periodontal surgery. No significant differences were found between the erratic and complete complier groups with regard to age, sex and treatment undertaken. Erratic complier patients lost more teeth than complete complier patients during supportive periodontal treatment. The main finding of this study was that no factors other than the complexity of active periodontal therapy had any influence on patients being compliant or non-compliant. Furthermore, no factors were found to affect the degree of compliancy. Our results suggest that patients' understanding of periodontal treatment plays a major role in compliance with supportive periodontal treatment. PMID- 7490606 TI - Relaxation effect of an audiovisual system on dental patients. Part 2. Palus amplitude. AB - Dental care services include relaxation of patients to reduce their displeasure at and tension during treatment. For this purpose, it is common to use audio or audiovisual programs, that is, to let them listen to music or watch TV. However, the former utilizes only the auditory sense, and the latter is disadvantageous in that a TV set is a rather large-scale unit. In the present study, we determined the relaxation effect of an audiovisual instrument on dental patients. The instrument, which gives virtual vision, was composed of a lightweight eyeset of the snow goggle type that has a built-in TV screen, with earphone and accessories. The relaxation effect was determined using a polygraph. PMID- 7490607 TI - Knowledge and attitudes of parents of children with nursing bottle syndrome in Serdang, Malaysia. AB - A study was conducted to investigate the relationship between the parents' level of knowledge, and their attitudes to nursing bottle syndrome. Of 284 children aged 2-6 years clinically examined, 153 were diagnosed as having nursing bottle syndrome based on the criterion that at least one upper maxillary incisor was affected by caries. The parents of the 153 affected children were then interviewed by questionnaire. The findings suggested that parents had adequate knowledge and a positive attitude towards maintaining satisfactory dental care for their children. However, these attitudes were not reflected in the dental health of their children. The implications of these findings are discussed. PMID- 7490608 TI - Evaluation of jawbone calcification in osteopetrotic (op/op) mice using computed tomography. AB - The study was conducted to evaluate the degree of calcification in the jawbones of 3 pairs of 4-week-old male op/op mice and littermates using computed tomography (CT) numbers. The mean CT numbers for op/op mice (n = 3) were 251.3 HU (range, 226 to 278) in the maxillary bone and 353.7 HU (range, 320 to 398) in the mandibular bone. The mean CT numbers for littermates were 336.7 HU (range, 324 to 350) and 596.6 HU (range, 580 to 604), respectively. The CT numbers of littermates were higher than in op/op mice for both maxillary and mandibular bones. These results indicated that the degree of bony calcification in littermates was higher than that in 4-week-old op/op mice in both the maxillary and mandibular bones. CT is able to demonstrate the degree of bony calcification in the jawbones of op/op mice using CT numbers, and it is suggested that CT is an extremely effective tool for examining the conditions of bone in vivo. This study also shows that op/op mice are useful for investigation of bone calcification or maturation. PMID- 7490609 TI - Morphometric analysis of epithelial components and dentinoid formation in non neoplastic calcifying odontogenic cyst. AB - Calcifying odontogenic cysts (COCs) represent a group of lesions that may be broadly classified into two main entities: cysts and neoplasms. In the present study 30 non-neoplastic cystic COCs were examined by a quantitative histological method in an attempt to calibrate the relative distribution of the type of epithelial lining, intensity of ghost cell formation and the amount of dentinoid present. The results showed that there are two main types of cystic COC: an odontoma-producing type and a non-odontoma-producing variant. Morphologically, tooth-like structures were a valid distinguishing feature, while morphometrically the odontoma-producing variant showed a greater amount of luminal and mural dentinoid as well as luminal ghost cells. Demographic analysis also revealed that the odontoma-producing COC occurred in younger patients and showed an even sex distribution, whereas the non-odontoma-producing type was seen in older patients and showed a predilection for females. Both subtypes were more prevalent in the Chinese population and occurred preferentially in the maxilla. PMID- 7490610 TI - An immunohistochemical study of the lining epithelium of odontogenic keratocyst. AB - The lining epithelium of 15 cases of odontogenic keratocyst (OKC) was evaluated immunohistochemically. The peroxidase-antiperoxidase technique was applied to study the distribution of polyclonal keratin and S-100 protein while the indirect method was used to examine monoclonal vimentin and desmin reactivity. Consistent positive keratin staining was revealed in the lining epithelium of all 15 OKCs with additional intense staining in the stratum corneum. None of the cases showed vimentin or desmin reactivity within the lining epithelium elements. One of the 15 cysts studied showed positive S-100 protein staining in the nuclei of the lining epithelial cells. The pertinent literature on the immunophenotyping of the lining epithelium of OKC is reviewed. PMID- 7490611 TI - Adsorption of saliva-coated and just-harvested Streptococcus sanguis to saliva coated hydroxyapatite beads. AB - It is possible that salivary proteins on bacterial cells have some influence on the adsorption of oral bacteria to the surface of oral tissues. The aim of this study was to clarify the influence of salivary proteins attached to the surface of Streptococcus sanguis ATCC 10556 (S. sanguis) on bacterial adsorption to hydroxyapatite beads treated with salivary proteins (S-HA beads). Just-harvested cells (N-system) and saliva-coated cells (C-system) were used. The results showed that adsorption to S-HA beads was gradually reduced by a covering of salivary proteins. No influence was observed on the adsorption of cells due to the presence of divalent cations, phosphate or EDTA when the N-system and C-system were compared. Though it was evident that hydrophobic bonding was involved in adsorption in the N-system, few such effects were observed in the C-system. Adsorption was inhibited to varying degrees by sugars in both systems. Pretreatment of cells with heat affected adsorption differently in the N-system and C-system. These findings suggest that salivary proteins attached to the bacterial cell surface may influence its adsorption to oral tissues. PMID- 7490612 TI - Two cases of symptomatic elongated styloid process. AB - Two cases of elongated styloid process are reported. The first occurred in a 27 year-old man whose only complaint was pain when moving the jaw, and its differential diagnosis was comparatively simple. The second case occurred in a 33 year-old woman, for whose condition differential diagnosis was more difficult, as she presented many symptoms, including neuralgic pain in the face and neck, pain in the pharynx when swallowing, and pain in the region of the temporomandibular joint, radiating to the subauricular and submandibular regions. Following surgical shortening of the elongated styloid process through an intraoral approach, both patients' symptoms disappeared or were improved. PMID- 7490613 TI - Intracranial cysts in autosomal dominant polycystic kidney disease. AB - Autosomal dominant polycystic kidney disease (ADPKD) is a systemic disorder well known for its association with intracranial aneurysms. A series of patients with ADPKD who were screened for the presence of an intracranial aneurysm were reviewed and found to include an unexpectedly high number with intracranial arachnoid cysts. Among 247 patients with ADPKD who underwent magnetic resonance imaging (180 cases) or high-resolution contrast-enhanced computerized tomography (67 cases), there were 151 women and 96 men with a mean age of 44 years. Intracranial arachnoid cysts were found in 20 patients (8.1%) with ADPKD compared to two (0.8%) in a control group without ADPKD matched for age, sex, and method of imaging (p < 0.0001). Multiple intracranial arachnoid cysts were found in two patients. Polycystic liver disease was present in 17 (85.0%) of the 20 patients with intracranial arachnoid cysts compared to 119 (52.4%) of the 227 patients without (p < 0.004). Pineal cysts were found in two patients (0.8%) and choroid plexus cysts were found in three patients (1.2%) but this was not different from the control population. None of the intracranial cysts was symptomatic and none was treated surgically. Intracranial arachnoid cysts are a relatively frequent incidental finding in patients with ADPKD, providing further support for the systemic nature of this disease. In the authors' experience with approximately 1500 patients with ADPKD, no complication has been encountered from an intracranial arachnoid cyst, suggesting that asymptomatic intracranial arachnoid cysts in patients with ADPKD require no treatment. PMID- 7490614 TI - Apoptosis of human glioma cells in response to calphostin C, a specific protein kinase C inhibitor. AB - Calphostin C acts at the regulatory domain as a highly selective inhibitor of protein kinase C (PKC), and staurosporine acts at the catalytic domain as a nonspecific PKC inhibitor. The authors investigated the capacity of calphostin C and staurosporine to promote apoptotic fragmentation of DNA in four human glioma cell lines. The exposure of glioma cell lines to 100 nM calphostin C for 2 to 8 hours induced a decrease in particulate PKC activities and exposure for 16 to 24 hours produced a concentration-dependent increase in internucleosomal DNA cleavage on agarose gel electrophoresis. In addition, the human glioma cells showed the classic morphological features of apoptosis: cell shrinkage, nuclear condensation, and the formation of apoptotic bodies. A 24-hour exposure to staurosporine failed to induce internucleosomal DNA fragmentation at concentrations generally used to achieve maximum inhibition of enzyme activity (50 nM) but promoted fragmentation at considerably higher concentration (more than 200 nM). Deoxyribonucleic acid fragments obtained from cells exposed to 100 nM calphostin C for 16 to 24 hours possessed predominantly 5'-phosphate termini, consistent with the action of a Ca++/Mg(++)-dependent endonuclease. Northern and Western blot analyses revealed that the exposure to 100 nM calphostin C for 4 hours failed to alter bcl-2 transcript and protein, but exposure for more than 8 hours decreased the amount of bcl-2 transcript and protein. Together, these observations suggest that calphostin C is capable of inducing apoptotic DNA fragmentation and cell death in a highly concentration dependent manner in human glioma cells and that the apoptosis is closely associated with the decrease in transcription and translation of bcl-2. PMID- 7490615 TI - Expression of bcl-2 in reactive and neoplastic astrocytes: lack of correlation with presence or degree of malignancy. AB - The bcl-2 protooncogene encodes a 26-kD protein that extends cell survival by blocking apoptosis. This protein has been found to be overexpressed in neoplastic neural cell lines, although its expression in reactive and neoplastic astrocytes in vivo has not been well characterized. The authors hypothesized that bcl-2 oncoprotein expression in gliomas might be positively correlated with the tumor's degree of malignancy. Sixty-three gliomas of various subtypes and histological grades were immunostained by bcl-2 protein and the percentage of positive cells was quantitatively assessed. All tumors contained neoplastic cells that were immunoreactive for the bcl-2 protein (range of cell positivity 1%-53%). It was found that bcl-2 expression did not vary significantly as a function of tumor subtype or grade (p < 0.1, one-way analysis of variance (ANOVA) on ranks) as compared to the cell proliferation marker Ki-67 (MIB-1) in which a very significant correlation with tumor grade was noted (p < 0.0000001, one-way ANOVA on ranks). In fact, the highest percentage of bcl-2 immunoreactive cells was noted in low-grade gliomas, that is, in juvenile pilocytic astrocytomas and oligoastrocytomas. The specificity of bcl-2 overexpression was also assessed in 10 nonneoplastic lesions associated with prominent reactive astrocytosis. In nine of these cases (90%), bcl-2-positive reactive astrocytes were observed, often in large numbers, whereas relatively few Ki-67 immunoreactive cells were noted. The authors conclude that bcl-2 oncoprotein expression as assessed immunohistochemically does not correlate with glial tumor type or grade and its overexpression is not confined only to neoplastic conditions. Instead, the finding of robust bcl-2 expression in low-grade glial tumors and in reactive astrocytes warrants the inference that resistance to apoptosis is a nonspecific finding in astrocytes associated with both reactive and neoplastic conditions. PMID- 7490616 TI - Magnetic resonance axonography of the rat spinal cord: postmortem effects. AB - The recent development of magnetic resonance (MR) axonography, which uses three dimensional anisotropy contrast (3DAC), a new algorithm for the treatment of an apparent diffusion tensor, has provided an unprecedented opportunity for visualizing the anatomical details of the spinal cord in live animals. In this study, the authors investigated the sensitivity of the 3DAC method in detecting pathological conditions by obtaining chronological MR axonography of the rat spinal cord immediately after induction of cardiac arrest. The results clearly demonstrated that 3DAC is highly sensitive to any perturbation of physiological conditions. Trichromatic coefficient analyses indicated postmortem changes observed pictorially are indeed due to loss of anisotropy. The study further indicated the presence of at least two independent factors responsible for observed physiological anisotropy. Considering its rather simple implementation process and high anatomical resolution as well as its sensitivity to pathological alteration, MR axonography based on the 3DAC method appears to be the ideal noninvasive imaging technique for assessment of the spinal cord in biomedicine. PMID- 7490617 TI - Doxorubicin encapsulated in sterically stabilized liposomes for the treatment of a brain tumor model: biodistribution and therapeutic efficacy. AB - Anthracyclines entrapped in small-sized, sterically stabilized liposomes have the advantage of long circulation time, reduced systemic toxicity, increased uptake into systemic tumors, and gradual release of their payload. To date, there is no information on the behavior of these liposomes in brain tumors. The objective of this study was to compare the biodistribution and clinical efficacy of free doxorubicin (F-DOX) and stealth liposome-encapsulated DOX (SL-DOX) in a secondary brain tumor model. Nine days after tumor inoculation Fischer rats with a right parietal malignant sarcoma received an intravenous dose of 6 mg/kg of either F DOX or SL-DOX for evaluation of drug biodistribution. For therapeutic trials a single dose of 8 mg/kg was given 6 or 11 days after tumor induction, or alternatively, weekly doses (5 mg/kg) were given on Days 6, 13, and 20. Liposome encapsulated DOX was slowly cleared from plasma with a t1/2 of 35 hours. Free-DOX maximum tumor drug levels reached a mean value of 0.8 microgram/g and were identical in the adjacent brain and contralateral hemisphere. In contrast, SL-DOX tumor levels were 14-fold higher at their peak levels at 48 hours, declining to ninefold increased levels at 120 hours. A gradual increase in drug levels in the brain adjacent to tumor was noted between 72 and 120 hours (up to 4 micrograms/g). High-performance liquid chromatography analysis identified a small amount of aglycone metabolites within the tumor mass from 96 hours and beyond, after SL-DOX injection. Cerebrospinal fluid levels were barely detectable in tumor-bearing rats treated with F-DOX up to 120 hours after drug injection (< or = 0.05 microgram/ml), whereas the levels found after SL-DOX were 10- to 30-fold higher. An F-DOX single-dose treatment given 6 days after tumor inoculation increased the rats' life span (ILS) by 135% over controls (p < 0.05) but was not effective if given on Day 11. In contrast, SL-DOX treatment resulted in an ILS of 168% (p < 0.0003) with no difference when given after 6 or 11 days. Treatment with three weekly doses of SL-DOX produced an ILS of 189% compared to 126% by F DOX (p < 0.0002). The authors conclude that the use of long-circulating liposomes as cytotoxic drug carriers in brain tumor results in enhanced drug exposure and improved therapeutic activity, with equal effectiveness against early small- and large-sized brain tumors. PMID- 7490618 TI - Prolonged survival of mice with glioma injected intracerebrally with double cytokine-secreting cells. AB - A novel approach toward the treatment of glioma was developed in a murine model. The genes for both interleukin-2 (IL-2) and interferon-gamma (IFN-gamma) were first transfected into a mouse fibroblast cell line that expresses defined major histocompatibility complex (MHC) determinants (H-2k). The double cytokine secreting cells were then cotransplanted intracerebrally with the Gl261 murine glioma cell line into syngeneic C57BL/6 mice (H-2b) whose cells differed at the MHC from the cellular immunogen. The results indicate that the survival of mice with glioma injected with the cytokine-secreting allogeneic cells was significantly prolonged, relative to the survival of mice receiving equivalent numbers of glioma cells alone. Using a standard 51Cr-release assay, the specific release of isotope from labeled Gl261 cells coincubated with spleen cells from mice injected intracerebrally with the glioma cells and the cytokine-secreting fibroblasts was significantly higher than the release of isotope from glioma cells coincubated with spleen cells from nonimmunized mice. The cellular antiglioma response was mediated by natural killer/lymphokine-activated killer and Lyt-2.2+ (CD8+) cells. The increased survival of mice with glioma and the specific immunocytotoxic responses after immunization with fibroblasts modified to secrete both IL-2 and IFN-gamma indicate the potential of an immunotherapeutic approach to gliomas with cytokine-secreting cells. PMID- 7490619 TI - Intracerebral infusion of thrombin as a cause of brain edema. AB - Purified thrombin from an exogenous source is a hemostatic agent commonly used in neurosurgical procedures. The toxicity of thrombin in the brain, however, has not been examined. This study was performed to assess the effect of thrombin on brain parenchyma, using the formation of brain edema as an indicator of injury. Ten microliters of test solution was infused stereotactically into the right basal ganglia of rats. The animals were sacrificed 24 hours later, and the extent of brain edema and ion content were measured. Concentrations of human thrombin as low as 1 U/microliter resulted in a significant increase in brain water content. Rats receiving 10 U/microliters had a mortality rate of 33% compared to no mortality in the groups receiving smaller doses. Thrombin-induced brain edema was inhibited by a specific and potent thrombin inhibitor, hirudin. A medical grade of bovine thrombin commonly used in surgery also caused brain edema when injected at a concentration of 2 U/microliters. Edema formation was prevented by another highly specific thrombin inhibitor, N alpha-(2-Naphthalenesulfonylglycyl)-4-DL phenylalaninepiperidid e (alpha-NAPAP). Thrombin-induced brain edema was accompanied by increases in brain sodium and chloride contents and a decrease in brain potassium content. Changes in brain ions were inhibited by both hirudin and alpha-NAPAP, corresponding to the inhibition of brain water accumulation. This study shows that thrombin causes brain edema when infused into the brain at concentrations as low as 1 U/microliter, an amount within the range of concentrations used for topical hemostasis in neurosurgery. PMID- 7490620 TI - Current concepts of brain edema. Review of laboratory investigations. AB - Klatzo's classification of brain edema into two types, vasogenic and cytotoxic, has been in general use since 1967. The former involves overall brain swelling due to fluid entry from the vasculature because of openings in the blood-brain barrier (BBB), whereas the latter refers to cell swelling without any loss of the normal impermeability of the BBB. This review principally covers new work that identifies the intracellular swelling of astrocytes as a major form of cytotoxic edema seen in many different kinds of brain injury. The term edema should be retained because of its familiarity; however, because such intracellular swelling is usually not a response to toxins, it is suggested that the term cellular edema is preferable to cytotoxic edema. The difficulties involved in measuring cellular edema clinically are discussed, and the belief that a "pure" form of either edema is unlikely to exist. It is emphasized that the mechanisms and direct consequences of vasogenic and cellular edema are so different that the connection is mainly semantic. Studies conducted in vitro have identified several potentially damaging secondary consequences of astrocytic swelling. One of the most important of these seems likely to be the increased release of excitatory amino acids from swollen astrocytes. Potential mechanisms for inhibition of the increased release of amino acids have been identified in vitro and could prove therapeutically useful. PMID- 7490621 TI - Brain edema and neurological status with rapid infusion of lactated Ringer's or 5% dextrose solution following head trauma. AB - Rapid infusion of 0.25 ml/g of 0.9% saline over 30 minutes has been shown to have no effect on electrolyte balance, neurological severity score (NSS), or brain edema, following closed head trauma (CHT). Rapid infusion of the same volume of 5% dextrose solution decreased blood sodium concentration, increased edema, and decreased NSS following CHT. In the present study the authors examined the effect of rapid infusion (30 minutes) of smaller volumes of 5% dextrose (0.08 ml/g and 0.16 ml/g) and of 0.25 ml/g lactated Ringer's solution on blood electrolyte concentrations, plasma osmolality, brain edema, and NSS. The purposes of this study were to determine whether rapid infusion of a large volume of lactated Ringer's solution could be given after CHT without increasing mortality or brain edema or producing electrolyte disturbances, and whether small volumes of 5% dextrose could be infused with few or none of the deleterious effects expected from large volumes of 5% dextrose. One hundred eighteen rats, which survived halothane anesthesia and CHT, were randomly assigned to one of 15 experimental groups. Fluids were administered beginning 1 hour after scalp incision or CHT. The NSS, extent of edema, blood electrolyte concentrations, and plasma osmolality in the groups treated with lactated Ringer's solution were not significantly different from those values in the nontreated groups. In addition, the mortality rate after CHT was not increased by administration of lactated Ringer's solution. The groups treated with 5% dextrose solution showed a significantly higher mortality rate, but the NSSs of the surviving rats were not different from controls. None of the groups treated with 0.16 ml/g 5% dextrose solution survived 24 hours. Although blood glucose concentration increased to 1126 +/- 102 g% (mean +/- standard deviation) and 1568 +/- 283 g% and blood sodium concentration decreased to 110.4 +/- 4.6 mEq/L and 92.0 +/- 5.2 mEq/L in the groups treated with 0.08 ml/g and 0.16 ml/g of 5% dextrose solution, respectively, plasma osmolality was normal and no significant difference could be found between the brain tissue specific gravity of animals in the nontreated and 5% dextrose treatment groups. It is concluded that in the CHT model used in this study, the large volume of lactated Ringer's solution did not affect blood electrolyte concentration, neurological outcome, or formation of brain edema, whereas smaller volumes of 5% dextrose solution increased blood glucose and decreased blood sodium concentrations, did not affect plasma osmolality, and had a deleterious effect on neurological outcome. PMID- 7490622 TI - Cardiovascular response to blood loss during high intracranial pressure. AB - The authors hypothesized that the combination of hemorrhage and increased intracranial pressure (ICP) has deleterious effects on cardiovascular function. The effect of blood loss during normal and increased ICP was studied in eight pigs. The mean arterial pressure (MAP), pulmonary arterial pressure, pulmonary capillary wedge pressure, cardiac output, and cerebrospinal fluid (CSF) pressure were measured. The regional tissue blood flow was determined with radioactive microspheres labeled with four different nuclides. High ICP (80% of MAP) was induced by infusion of artificial CSF into the cisterna magna. The response to rapid arterial bleeding of 25% of blood volume was measured. The decrease in blood flow to the intestine, skeletal muscle, and the kidneys after blood loss was significantly greater during high ICP. The decrease in blood flow to the spleen and pancreas tended to be greater during high ICP, whereas the changes in blood flow to the liver, adrenal glands, and heart muscle showed no such tendency. The fall in cardiac output and heart stroke volume after blood loss were more pronounced when the ICP was high, and the increase in systemic vascular resistance was considerably greater. These observations suggest that during high ICP the physiological protective mechanisms against blood loss are impaired in the systemic circulation, and a loss of 25% of the blood volume, normally well compensated for, may induce a state of shock. PMID- 7490624 TI - Lymphoplasmacyte-rich meningioma in a child. Case report. AB - A rare case of lymphoplasmacyte-rich meningioma observed in a young girl is reported. The first clinical manifestations of the disease were seizures. Neuroradiological images favored the existence of a meningioma. Abnormalities in the patient's blood chemistry, principally including hypergammaglobulinemia and inflammatory syndrome, were associated with the disease. The tumor was histologically confirmed as meningioma with massive infiltrates of type B lymphocytes. The pathophysiology of the conspicuous lymphoplasmacyte infiltrates, responsible for peripheral blood abnormalities, has remained poorly understood. Alternative diagnostic hypotheses of masses that mimic this type of meningioma are discussed. PMID- 7490623 TI - Brainstem tethering in Dandy-Walker syndrome: a complication of cystoperitoneal shunting. Case report. AB - Treatment of the Dandy-Walker syndrome has included placement of a ventriculoperitoneal shunt alone or in combination with a posterior fossa cystoperitoneal shunt. Complications in shunting are common and are usually related to malfunction or infection. The authors present a case in which the patient developed headaches and focal cranial nerve deficits following infection caused by a cystoperitoneal shunt. Magnetic resonance imaging showed tethering of the brainstem. A posterior fossa craniotomy with microsurgical untethering and cyst fenestration achieved two goals: improvement of the focal cranial nerve deficits and elimination of the cystoperitoneal shunt. PMID- 7490625 TI - Huge Langerhans cell histiocytosis granuloma of choroid plexus in a child with Hand-Schuller-Christian disease. Case report. AB - In the literature, only two previous cases of presumed Langerhans cell histiocytosis (LCH) granuloma involving choroid plexus have been reported in patients with Hand-Schuller-Christian (HSC) disease, but those cases were not verified by immunohistochemistry or electron microscopy. The authors report a case of LCH granuloma of the choroid plexuses of bilateral lateral ventricles and the fourth ventricle in a child affected by HSC disease. This disease was confirmed by the presence of Birbeck granules on electron microscopy and positive reactivity to S-100 protein. Clinical, radiographic, and histological features are discussed. This case illustrates that LCH granuloma should be considered in the differential diagnosis of intraventricular masses. PMID- 7490626 TI - Gradual resolution of an arachnoid cyst after spontaneous rupture into the subdural space. Case report. AB - The authors present the case of a young patient in whom the spontaneous rupture of a known, long-standing middle fossa arachnoid cyst into the subdural space was documented. This spontaneous rupture was followed by gradual diminution of the cyst to near-complete resolution. PMID- 7490627 TI - False-positive inferior petrosal sinus sampling in the diagnosis of Cushing's disease. Report of two cases. AB - Inferior petrosal sinus sampling (IPSS) for adrenocorticotropic hormone (ACTH) levels in patients with Cushing's syndrome has become a useful method to distinguish ACTH-secreting pituitary tumors (Cushing's disease) from other causes of the syndrome, principally ectopic adrenocorticotropin secretion by an occult tumor. Although the test is generally regarded as highly specific, the authors recently encountered two patients whose IPSS measurements were false-positive for Cushing's disease. The results of IPSS suggested a pituitary origin of ACTH secretion in both patients, but transsphenoidal surgery failed to disclose a pituitary adenoma or to improve postoperative plasma cortisol levels. Both patients subsequently were found to have an ACTH-secreting carcinoid tumor of the lung. The false-positive IPSS studies were due to periodic hormonogenesis. The patients must be hypercortisolemic at the time IPSS is performed for the study to be valid. PMID- 7490628 TI - Ectopic pituitary adenoma occurring in the interpeduncular cistern. Case report. AB - Intracranial ectopic pituitary adenoma occurs most frequently in the suprasellar cistern, usually in continuity with the pituitary stalk. Such tumors probably originate from cells of the pars tuberalis located above the diaphragma sellae or from aberrant anterior pituitary cells of the pituitary stalk. The authors report the case of a 37-year-old woman with Cushing's syndrome caused by an ectopic pituitary adenoma of unique location: the tumor was separate form the pituitary stalk and confined within the interpeduncular cistern. After surgical removal of the tumor, continued improvement in the patient's laboratory results and disappearance of her endocrine symptoms strongly indicated the absence of adenoma cells in the pituitary gland or stalk. The tumor in the present case appears to have arisen from aberrant pituitary cells that were present in the leptomeninges of the basal surface of the hypothalamus. PMID- 7490629 TI - Cannulated screws for odontoid screw fixation and atlantoaxial transarticular screw fixation. Technical note. AB - Cannulated screw systems use thin Kirschner wires (K-wires) that have been drilled into the bone to direct screw trajectories accurately into small bone fragments. Use of the K-wires avoids overdrilling the pilot holes and allows fixation of adjacent bone fragments during screw insertion. Hollow tools and hollow screws are inserted into the bone over the K-wires. Cannulated screw fixation is useful in the cervical spine to stabilize odontoid fractures and to treat atlantoaxial instability. This report describes techniques for successful cannulated screw insertion and methods to minimize complications. Cannulated screws have several distinct advantages compared to noncannulated screws: 1) the K-wires guide the screw position into the bone; 2) the K-wire trajectory can be repositioned easily if the original trajectory was not ideal; 3) the K-wires allow continuous fixation of adjacent unstable bone fragments; and 4) the K-wires prevent migration of unstable bone fragments during screw insertion. Complications associated with the K-wire (breakage, repositioning, and advancement) can be minimized using precise operative techniques, a specialized tool system, and intraoperative fluoroscopic monitoring. A unique cannulated screw tool system was developed specifically for upper cervical fixation to allow percutaneous drilling using long tunneling devices, tissue sheaths, drill guides, and long K-wires. These tools allow delivery of cannulated fracture-fixation screws at a low angle to the spine through long soft-tissue trajectories. Cannulated screws have significant advantages compared to noncannulated screws for fixation of the unstable cervical spine. PMID- 7490630 TI - Safe removal of retained ventricular catheters using intraluminal choroid plexus coagulation. Technical note. AB - The authors describe a simple technique that reduces the incidence of retained ventricular catheters and minimizes the risk of life-threatening intraventricular hemorrhage during the removal of an obstructed, adherent ventricular catheter in patients with hydrocephalus. The technique requires no special equipment and has been successfully used, without complications, in a prospective series of 12 patients with 13 blocked, adherent ventricular catheters. PMID- 7490632 TI - Near-infrared spectroscopy. PMID- 7490631 TI - A tribute to Dr. A. Earl Walker. AB - Doctor Walker died on January 1, 1995, at the age of 87 years. His contributions to neuroscience and neurosurgery are presented and discussed, together with his impact on national and international organizations of neurosurgery. A short account of his background and professional career is given, with emphasis on his 25 years at The Johns Hopkins Hospital as Chief of Neurosurgery. PMID- 7490633 TI - Shunt obstruction and CSF edema. PMID- 7490634 TI - Hyperventilation and head injury. PMID- 7490635 TI - Hyperventilation and head injury. PMID- 7490636 TI - Hyperventilation and head injury. PMID- 7490637 TI - Cortical stimulation for central pain. PMID- 7490638 TI - Cerebral perfusion pressure: management protocol and clinical results. AB - Early results using cerebral perfusion pressure (CPP) management techniques in persons with traumatic brain injury indicate that treatment directed at CPP is superior to traditional techniques focused on intracranial pressure (ICP) management. The authors have continued to refine management techniques directed at CPP maintenance. One hundred fifty-eight patients with Glasgow Coma Scale (GCS) scores of 7 or lower were managed using vascular volume expansion, cerebrospinal fluid drainage via ventriculostomy, systemic vasopressors (phenylephrine or norepinephrine), and mannitol to maintain a minimum CPP of at least 70 mm Hg. Detailed outcomes and follow-up data bases were maintained. Barbiturates, hyperventilation, and hypothermia were not used. Cerebral perfusion pressure averaged 83 +/- 14 mm Hg; ICP averaged 27 +/- 12 mm Hg; and mean systemic arterial blood pressure averaged 109 +/- 14 mm Hg. Cerebrospinal fluid drainage averaged 100 +/- 98 cc per day. Intake (6040 +/- 4150 cc per day) was carefully titrated to output (5460 +/- 4000 cc per day); mannitol averaged 188 +/ 247 g per day. Approximately 40% of these patients required vasopressor support. Patients requiring vasopressor support had lower GCS scores than those not requiring vasopressors (4.7 +/- 1.3 vs. 5.4 +/- 1.2, respectively). Patients with vasopressor support required larger amounts of mannitol, and their admission ICP was 28.7 +/- 20.7 versus 17.5 +/- 8.6 mm Hg for the nonvasopressor group. Although the death rate in the former group was higher, the outcome quality of the survivors was the same (Glasgow Outcome Scale scores 4.3 +/- 0.9 vs. 4.5 +/- 0.7). Surgical mass lesion patients had outcomes equal to those of the closed head-injury group. Mortality ranged from 52% of patients with a GCS score of 3 to 12% of those with a GCS score of 7; overall mortality was 29% across GCS categories. Favorable outcomes ranged from 35% of patients with a GCS score of 3 to 75% of those with a GCS score of 7. Only 2% of the patients in the series remained vegatative and if patients survived, the likelihood of their having a favorable recovery was approximately 80%. These results are significantly better than other reported series across GCS categories in comparisons of death rates, survival versus dead or vegetative, or favorable versus nonfavorable outcome classifications (Mantel-Haenszel chi 2, p < 0.001). Better management could have improved outcome in as many as 35% to 50% of the deaths. PMID- 7490639 TI - Near-infrared spectroscopy use in patients with head injury. AB - A multimodality recording system was used in 14 ventilated patients with closed head injury to assess the potential use of near-infrared spectroscopy (NIRS) in the neurointensive care unit. Signals of intracranial pressure, cerebral perfusion pressure, peripheral oxygen saturation, jugular venous saturation, and NIRS-derived changes in the chromophores of oxy- and deoxyhemoglobin were digitized and recorded. After a review of 886 hours of continuous monitoring, 376 hours were considered free from artifact and were entered for final analysis. In nine of the patients 38 events were recorded that demonstrated clear changes in cerebral perfusion pressure accompanied by hemodynamic changes in middle cerebral artery flow velocity (transcranial Doppler) and cortical perfusion (laser Doppler flowmetry). Near-infrared spectroscopy showed correlated changes in 37 events (97%) whereas jugular venous saturation monitoring registered only 20 (53%). There was associated peripheral oxygen desaturation in eight cases (21%), intracranial hypertension in 10 (26%), and cerebral hyperemia in eight (21%). The remaining 12 events (32%) appeared to be complex changes of uncertain origin. Iatrogenic factors were identified as causative in 14 cases (37%). The potential application of NIRS in adults and the importance of using multiple parameter recording systems in the interpretation of cerebral events are discussed. PMID- 7490640 TI - The transfacet pedicle-sparing approach for thoracic disc removal: cadaveric morphometric analysis and preliminary clinical experience. AB - A number of operative techniques have been described for the treatment of herniated thoracic discs. The transfacet pedicle-sparing approach allows for complete disc removal with limited spinal column disruption and soft-tissue dissection. Fifteen cadaveric spinal columns were used for evaluation of exposure, development of thoracic microdiscectomy instrumentation, and establishment of morphometric measurements. This approach was used to remove eight thoracic discs in six patients. Levels of herniation ranged from T-7 through T-11. Preoperatively, all patients had moderate to severe axial pain, and three (50%) of the six had radicular pain. Myelopathy was present in four (67%) of the six patients. Through a 4-cm opening, the ipsilateral paraspinal muscles were reflected, and a partial facetectomy was performed. The disc was then removed using specially designed microscopic instrumentation. Postoperatively, the radiculopathy resolved in all patients. Axial pain and myelopathy were completely resolved or significantly improved in all patients. The minimal amount of bone resection and muscle dissection involved in the operation allows for: 1) decreased operative time and blood loss; 2) diminished perioperative pain; 3) shorter hospitalization time and faster return to premorbid activity; 4) avoidance of closed chest tube drainage; and 5) preservation of the integrity of the facet-pedicle complex, with potential for improvement in outcome related to axial pain. This technique appears best suited for the removal of all centrolateral discs, although it has been used successfully for treating a disc occupying nearly the entire ventral canal. The initial experience suggests that this approach may be used to safely remove appropriately selected thoracic disc herniations with good results. PMID- 7490642 TI - Age-related multifactorial causes of neurological deterioration after early surgery for aneurysmal subarachnoid hemorrhage. AB - The factors contributing to neurological deterioration after early surgery for aneurysmal subarachnoid hemorrhage (SAH) were investigated. One hundred forty-two patients who underwent surgery within 3 days after SAH and recovered consciousness were divided into three age groups: 49 years of age or younger (Group A), 50 to 64 years old (Group B), and 65 years of age or older (Group C). Among these, 40 patients (28%) overall showed neurological deterioration; these cases were analyzed in detail. Although the highest incidence of deterioration was noted in patients in Group C (42%), angiographic vasospasm, quantified by measuring the change in the ratio of the diameters of the intracranial arteries to the extracranial internal carotid artery, was negatively correlated with age. In elderly patients, the severity of angiographic vasospasm was not related to the reversibility of symptoms or the outcome. At the time of aggravation, associated systemic complications such as cardiac decompensation, hypoxia, and electrolyte imbalance were noted in two (18%) of 11 patients in Group A, five (38%) of 13 in Group B, and eight (50%) of 16 in Group C, and these complications were significantly correlated with poor outcome in Group C. Although arterial narrowing is a leading cause of neurological deterioration after early aneurysmal surgery, the etiology is often multifactorial, especially in elderly patients, suggesting that hypervolemic therapy, which might provoke various complications, should be performed carefully under intensive monitoring. PMID- 7490641 TI - Surgical approaches for the correction of unstable thoracolumbar burst fractures: a retrospective analysis of treatment outcomes. AB - The authors retrospectively studied 49 nonparaplegic patients who sustained acute unstable thoracolumbar burst fractures. All patients underwent surgical treatment and were followed for an average of 27 months. All but one patient achieved solid radiographic fusion. Three treatment groups were studied: the first group of 16 patients underwent anterior decompression and fusion with instrumentation; the second group of 27 patients underwent posterior decompression and fusion; and the third group of six patients had combined anterior-posterior surgery. Prior to surgical intervention, these groups were compared and found to be similar in age, gender, level of injury, percentage of canal compromise, neurological function, and kyphosis. Patients treated with posterior surgery had a statistically significant diminution in operative time and blood loss and number of units transfused. There were no significant intergroup differences when considering postoperative kyphotic correction, neurological function, pain assessment, or the ability to return to work. Posterior surgery was found to be as effective as anterior or anterior-posterior surgery when treating unstable thoracolumbar burst fractures. Posterior surgery, however, takes the least time, causes the least blood loss, and is the least expensive of the three procedures. PMID- 7490643 TI - A prospective 15-year follow up of 154 consecutive patients with trigeminal neuralgia treated by percutaneous stereotactic radiofrequency thermal rhizotomy. AB - There is a lack of prospective studies for the long-term results of percutaneous stereotactic radiofrequency rhizotomy (PSR) in the treatment of patients with trigeminal neuralgia. The authors present results in 154 consecutive patients with trigeminal neuralgia treated by PSR and prospectively followed for 15 years. Ninety-nine percent of the patients obtained initial pain relief after one PSR. Dysesthesia occurred in 31 patients (23%): in 7% with mild initial hypalgesia; in 15% with dense hypalgesia; and in 36% with analgesia. Dysesthesia was mild and did not require treatment in most patients. The corneal reflex was absent or depressed in 29 patients, and keratitis developed in three patients. In 19 of 22 patients with trigeminal motor weakness, the paresis resolved within 1 year. Of 33 patients who had pain recurrence, 10 patients had pain that was mild or controlled with medications, and 23 patients required additional surgical treatment. The authors estimated using Kaplan-Meier analysis that the 14-year recurrence rate was 25% in the total group: 60% in patients with mild hypalgesia, 25% in those with dense hypalgesia, and 20% in those with analgesia. Timing of pain recurrence varied according to the degree of sensory loss. All pain recurrences in patients with mild hypalgesia occurred within 4 years after surgery; 10% more of the patients with dense hypalgesia had pain recurrences within the first 10 years compared with patients with analgesia. The median pain free survival rate was 32 months for patients with mild hypalgesia and more than 15 years for patients with either analgesia or dense hypalgesia. Of the 100 patients followed for 15 years after one or two PSR procedures, 95 patients (95%) rated the procedure excellent (77 patients) or good (18 patients). The authors conclude that PSR is an effective, safe treatment for trigeminal neuralgia. Dense hypalgesia in the painful trigger zone, rather than analgesia, should be the target lesion. PMID- 7490644 TI - Proximal-to-distal facial amplitude ratios as predictors of facial nerve function after acoustic neuroma excision. AB - Electrophysiological studies (for example, electroneuronography, nerve action potentials, absolute amplitudes of the muscle compound action potentials, and stimulation thresholds) do not accurately predict facial nerve function after the excision of acoustic neuromas. To eliminate individual nerve variability, the authors measured the ratio of the amplitudes of muscle compound action potentials produced by stimulating the facial nerve at the brainstem proximally and at the internal auditory meatus near the transverse crest distally after total tumor excision in 20 patients. The mean tumor size was 36 mm. The facial nerves were anatomically intact in all patients after tumor excision. The follow-up period ranged from 14 to 28 months. Facial nerve outcome was determined by a modified House-Brackmann grading scale. Initial facial nerve function was measured at Days 4 to 7 postoperatively, and final function was the grade at last follow up. The following results were obtained: all patients with proximal-to-distal amplitude ratios greater than 2:3 had Grade III or better initial function and Grade I final facial nerve function; 90% of patients with amplitude ratios between 1:3 and 2:3 had Grade III or worse initial facial nerve function, and 100% of these patients had Grade III or better final facial nerve function; all patients with amplitude ratios less than 1:3 had Grade IV or worse initial and final facial nerve function. The authors conclude that the proximal-to-distal amplitude ratios after acoustic neuroma excision can accurately predict postoperative facial nerve function. PMID- 7490645 TI - The incidence of primary central nervous system neoplasms before and after computerized tomography availability. AB - The observed incidence of primary intracranial neoplasms has increased dramatically among the elderly over the past several years. It is unclear to what extent this increase can be attributed to improved detection by new imaging technology and what portion might represent a true increase in incidence. The first computerized tomography (CT) scanner was operative in Norway in 1975 and became available in all geographical regions within 4 years. The author analyzed incidence data from the population-based Cancer Registry of Norway, placing particular emphasis on the basis for diagnosis (histological, imaging, or clinical findings) for the registrations. In the decade from 1983 to 1992, when the use of CT was widespread, incidence rates of primary intracranial neoplasms for individuals aged 55 to 74 and above 74 years were, respectively, 1.76 (95% confidence interval (CI), 1.63-1.90) and 3.35 (95% CI, 2.70-4.15) times the rates in 1963 to 1972 before CT was available. For persons older than 74 years, the relative incidence increase was 1.98-fold (95% CI, 1.53-2.56) for histologically verified tumors; 15.20 (95% CI, 7.80-29.61) when based on imaging (without histological evidence); and 3.33 (95% CI, 1.93-5.76) when based on clinical diagnosis (without imaging or histological evidence). In this age group, the increase for gliomas was 1.77 (95% CI, 1.18-2.65) and 2.28 (95% CI, 1.53-3.39) for meningioma patients. The present study demonstrates that new imaging technology has improved the diagnostic yield of primary intracranial neoplasms. For those older than 74 years, a large part of the three-fold incidence increase from the pre-CT decade 1963 to 1972 to the post-CT decade 1983 to 1992 can be attributed to improved tumor detection. PMID- 7490647 TI - Otoplasty for correction of the prominent ear. AB - The oral and maxillofacial surgeon is aware of the need to evaluate and treat the entire facial complex. To gain maximum esthetic results, excessive ear prominence occasionally needs to be addressed. Excessive ear prominence can be the result of failure of scapha folding, conchal hypertrophy, conchal malposition, or a combination of these deformities. Management of this problem is based on the accurate diagnosis of the deformity and a thorough understanding of the basic techniques that address them. This article presents an overview of otoplasty for the treatment of prominent ears. PMID- 7490646 TI - In vitro comparison of parameters affecting the fixation strength of sagittal split osteotomies. AB - PURPOSE: The goal of this study was to determine how different parameters affect the bending strength of human cadaver mandibles that have undergone a sagittal split osteotomy. MATERIALS AND METHODS: The effects of screw material (titanium [Ti] vs polylactic acid/polyglycolic acid [PLA/PGA]), screw configuration (linear vs inverted L-shape), screw diameter (2.0 mm vs 2.7 mm), material into which screws were inserted (human mandible, bovine rib, synthetic polymer), and loading rate (1.0 mm/min vs 10.0 mm/min) were quantified. Also, biomechanical principles were used to model shear stress and displacement. Variable lever arms, screw material, screw diameter, screw configuration, distance between screws, and bone properties were all evaluated in this model. RESULTS: Accounting for variable mandible geometries and differentiating between deflections (and shear stresses) due to bending and due to torsion, in vitro mechanical testing revealed that there was a statistically significant difference in total shear stress at 3 mm of deflection depending on screw material (Ti > PLA/PGA), screw diameter, and material into which screws are inserted (mandibles > ribs = synthetic polymer). There was no significant difference in total shear stress depending on screw configuration or strain rate. CONCLUSION: Total shear stress and deflections are important and more viable parameters than load to assess parameters of clinical importance in osteotomy or fracture fixation. PMID- 7490648 TI - Application of the interseptal alveolotomy for closing the oroantral fistula. AB - PURPOSE: This article describes a new application of the interseptal alveoplasty to close oroantral fistulae caused by dental extraction. The surgical technique is described, and its advantages are compared with those of other techniques. MATERIALS AND METHODS: The technique was used to treat eight patients with comparatively small fistulae in the soft tissue, approximately 1 to 5 mm wide. RESULTS: All fistulae were successfully closed. CONCLUSION: It is proposed that the technique provides greater flexibility in the treatment of fistulae. In addition, because there is a bony base, this promotes more successful healing than other commonly used procedures. PMID- 7490649 TI - Lag-screw osteosynthesis of mandibular condyle fractures: a clinical and radiological study. AB - PURPOSE: This study evaluates the clinical and radiologic results after open reduction and lag-screw osteosynthesis of fractured mandibular condyles. MATERIALS AND METHODS: Eleven adult patients underwent surgery for displaced or dislocated mandibular condyle fractures via a submandibular approach. The repositioned fragments were fixed using lag screws designed by Krenkel or Eckelt. Maxillomandibular fixation was used postoperatively for 2.6 weeks on average (range, 1 to 4 weeks) in nine patients. RESULTS: Slight transient weakness of the mandibular branch of the facial nerve occurred in three cases. Occlusal adjustment was needed in another three cases. Radiologically, the fracture line disappeared at 22.5 weeks on average (range, 8 to 38 weeks). Three screws had to be removed because of loosening. There were signs that the screws had migrated caudally from their original position in seven cases. Twenty-one months on average after operation (range, 8 to 31 months), patients were satisfied with treatment. Clinically, all patients had a stable occlusion and symmetry of the face. All had greater than 5-mm symmetrical lateral jaw excursions. Ten patients had wide (> 40 mm) painless mouth opening. Healing in malposition occurred in four cases and there was considerable shortening of the mandibular ramus in four cases. CONCLUSIONS: Despite good clinical results, lag screws do not meet the needs for rigid internal fixation in the treatment of mandibular condyle fractures. PMID- 7490650 TI - The stability of segmentalized Le Fort I osteotomies with miniplate fixation in patients with maxillary hypoplasia. AB - PURPOSE: This study evaluates the postsurgical stability of segmentalized Le Fort I osteotomies with miniplate fixation in patients with maxillary hypoplasia. MATERIALS AND METHODS: Eighteen adult patients who had undergone segmentalized Le Fort I osteotomy for anterior and inferior repositioning of their hypoplastic maxilla were studied radiographically. The positional change of five maxillary landmarks (PNS, ANS, A, U1, and UM) were measured on serial cephalometric radiographs according to a reference frame constructed by using the SN line for superimposition, sella as the origin of the coordinates, and a line parallel to Frankfort horizontal plane as the x-axis. The positional changes of all variables were measured twice, and the means were calculated for analysis. The paired t test was used for statistical analysis. RESULTS: Statistically significant mean vertical change (P < .01) was found in the anterior maxilla, with U1 having moved up by 27.8% (1.5 mm) of its initial downward surgical movement by 1 year postoperatively. U1 and UM had moved forward 2.0 mm and 1.5 mm, respectively (P < .01) by 1 year postoperatively, but the bony landmarks had no statistically significant change in their horizontal positions. CONCLUSION: Although statistically significant postsurgical changes were found at 1 year postoperatively, overcorrection is not recommended because of the large individual variation observed and the relatively small magnitude of the relapse. PMID- 7490651 TI - Maxillomandibular advancement surgery in obstructive sleep apnea syndrome patients: long-term surgical stability. AB - PURPOSE: This study examined the long-term skeletal stability of relatively large maxillomandibular advancement surgery in the treatment of obstructive sleep apnea syndrome (OSAS). MATERIALS AND METHODS: The presurgical, immediate (< 1 month), and long-term (> 12 months) postsurgical cephalometric radiographs of 19 patients undergoing maxillomandibular advancement with simultaneous genioplasty for OSAS were studied. The cephalometric measurements and calculations were performed using a commercial cephalometric software. RESULTS: Maxillary and mandibular advancement seems to be stable in the horizontal and vertical planes over the long term. There was no significant correlation between the amount of surgical advancement and the amount of postsurgical instability, with the exception of gonion in the vertical plane. CONCLUSION: The results from this study indicate that large surgical advancements in OSAS patients result in relatively stable repositioning of the maxilla and mandible over the long term. PMID- 7490652 TI - Ultrasonographic anatomy of the anterior neck: implications for tracheostomy. AB - PURPOSE: Tracheostomy is the preferred intervention for surgical establishment of an upper airway in the treatment of certain elective maxillofacial procedures. Complication rates associated with surgical tracheostomy are largely related to the restricted possibilities in diagnosis of anatomic variations. Therefore, endoscopic-guided tracheostomy is increasingly being used to enhance precision and safety. The purpose of this study was to assess the ultrasonographic anatomy of the anterior neck with regard to the performance of tracheostomy. PATIENTS AND METHODS: Fifty patients underwent ultrasonographic investigation to analyze the relationship of anatomic structures lying in the immediate vicinity of the tracheostomy site. RESULTS: The data presented demonstrate the feasibility and advantage of preoperative ultrasonography to visualize the structures of the infrahyoid region. CONCLUSION: Preoperative ultrasonography may be of great importance in diagnosing individual anatomic variations at the site of tracheostomy and in avoiding complications. PMID- 7490653 TI - Wound healing after multisegmental Le Fort I osteotomy and transection of the descending palatine vessels. AB - PURPOSE: Vascular ischemia has been associated with improper soft tissue flap design, stretching of the palatal vascular pedicle, bony segmentation, transection of the descending palatal vessels, or hypotension. This study examined Le Fort I osteotomy wound healing after some of these surgical maneuvers. MATERIALS AND METHODS: Clinically analogous four-segment Le Fort I osteotomies were accomplished through circumvestibular incisions in nine adult rhesus monkeys and the animals were killed at 0, 3, 7, 14, and 28 days after surgery. Revascularization and bone healing were studied by microangiographic and histologic techniques. RESULTS: The findings indicated that the palatal mucosa or labial-buccal gingiva and mucosa provide adequate nutrient pedicles for Le Fort I osteotomies accomplished through a circumvestibular type incision. CONCLUSIONS: It was concluded that segmentalization, stretching of the vascular pedicles, or transection of the descending palatine vessels have only transitory discernible effects on revascularization and bone healing. PMID- 7490654 TI - The effects of intra-articular deposition of betamethasone in the goat temporomandibular joint. AB - PURPOSE: This study evaluated the response of the articular surfaces of the normal goat temporomandibular joint (TMJ) to intra-articular injections of betamethasone suspension. METHODS: Thirty female goats were divided into four experimental groups of seven each and one control group of two animals. The design resulted in 24 joints receiving from one to nine weekly injections of betamethasone suspension, 0.085 mg/kg. The 24 contralateral joints received an identical array of saline injections. In the remaining 12 joints, four were unilaterally injected with saline, four were unilateral uninjected joints, and four were bilateral uninjected joints. All of the joints were inspected grossly and histologically assessed for any intra-articular changes associated with corticosteroid injections. RESULTS: Comparative examination of the gross and histologic features of the injected joints and the eight normal joints showed no significant adverse effects on nondiseased TMJs. CONCLUSION: Intra-articular injection of betamethasone suspension at the dosage and delivery rate used had no harmful effects on the TMJs of nondiseased adult female goats. Because of possible differences between species, these results should not be taken as an indication that betamethasone suspension will not have adverse effects in the human TMJ. These findings support the need for further investigations into the physical and biochemical responses of normal and diseased articular fibrocartilage to different glucocorticosteroids at various concentrations and delivery rates using the goat and other animal models. PMID- 7490655 TI - The potential risks, complications, and prevention of deep vein thrombosis in oral and maxillofacial surgery patients. AB - PURPOSE: Deep vein thrombosis is a complication in surgical patients with a potential for disastrous results. This article discusses the pathogenesis, prevention, and treatment of this condition. CONCLUSION: Surgeons should be acutely aware of the potential development of deep vein thrombosis and should take prophylactic measures to prevent this problem as part of their surgical routine. PMID- 7490656 TI - Molecular biology of temporomandibular joint disorders: proposed mechanisms of disease. AB - PURPOSE: The biologic processes of temporomandibular joint adaptation and disease are poorly understood. However, recent technologic advances have provided methods that allow sophisticated studies of the molecular mechanisms that are relevant to the pathophysiology of degenerative temporomandibular joint diseases. This review examines current models of the molecular events that may underlie both adaptive and pathologic responses of the articular tissues of the temporomandibular joint to mechanical stress. It is hoped that an increased understanding of these complex biologic processes will lead to improved diagnostic and therapeutic approaches directed to the management of temporomandibular disorders. PMID- 7490657 TI - Simultaneous malignant melanoma and squamous cell carcinoma of the oral cavity: a case report. AB - Primary malignant melanoma of the oral cavity is rare. The incidence of oral melanoma constitutes between 0.2% and 8.0% of all melanomas, although higher percentages are reported in Japanese and other nonwhite races. In contrast, squamous cell carcinoma is the most common malignant neoplasm in the oral region. Multiple cancers may develop independently over the course of time in one organ, or double cancers may develop separately in different organs. Both types of cancer are collectively referred to as multiple primary cancers. The simultaneous occurrence of two histologically distinct primary malignancies in the same organ is extremely rare. We present a case of simultaneous malignant melanoma in association with squamous cell carcinoma of the oral cavity. PMID- 7490658 TI - Basal cell adenocarcinoma of the parotid gland: a case report with immunohistochemical, ultrastructural findings and review of the literature. PMID- 7490660 TI - Orbital pseudotumor: report of a case after treatment for temporomandibular joint dysfunction. PMID- 7490659 TI - Massive osteolysis of the mandible with subsequent obstructive sleep apnea syndrome: a case report. PMID- 7490661 TI - Malignant and premalignant change in odontogenic cysts. PMID- 7490662 TI - Autotransplantation in cleidocranial dysplasia: case report with 5-year follow up. PMID- 7490663 TI - Intraoral removal of the enlarged mandibular angle associated with masseteric hypertrophy. PMID- 7490664 TI - A technique for ensuring accurate bone cuts in the intraoral vertical ramus osteotomy. PMID- 7490665 TI - Reorganization of the annual meeting. PMID- 7490666 TI - Comparison of partial and full crowns as retainers in the same bridge. AB - The purpose of this study was to assess longitudinally the success of partial and full crown abutment pairs in the same bridges. Fifty patients with a mean age of 45 years (range 22-70 years) were examined during spring 1981 (1-5 year follow up), 33 of whom were examined a second time during spring 1990 (10-15 year follow up). The status of the partial crowns, full crowns, and their abutment teeth was assessed using standardized questions, and clinical and radiographic examinations. At the first follow-up examination, all partial and full crowns, and consequently all bridges, were still in use. Before the second follow-up examination one of the full crown abutments had become badly decayed, and the bridge, including the partial crown, had been removed. The failure rate was 0.2% per year for the full crown as a unit, and consequently 0.2% per year for the bridge as a unit. PMID- 7490667 TI - Radiographic amelocemental junction and alveolar crest: effect of X-ray beam angulation. AB - The effect of X-ray beam angulation on the radiographic images of the amelocemental junction (ACJ) and alveolar crest (AC) and their positions relative to one another were assessed using a new laboratory model for radiographic studies. A total of 39 teeth with 62 usable approximal surfaces were included in the investigation. When all tooth types were combined, changes in vertical or horizontal angulation from baseline through 20 degrees had no significant effect on the distance between the marked (true) ACJ and marked (true) AC. However, the distance between the apparent (radiographic) ACJ and apparent (radiographic) AC was significantly reduced by varying the beam angulation in either a vertical or horizontal direction. Results for individual tooth types were qualitatively similar but larger discrepancies were found in molars followed by premolars and incisors. The results emphasize the importance of maintaining the ideal X-ray beam, tooth, bone, film relationship when taking radiographs for the assessment of periodontal destruction. PMID- 7490668 TI - The effect of glaze on restorative glass-ionomer cements: evaluation of environmental durability in lactic acid solution. AB - The surface texture changes of three kinds of glass-ionomer cements were compared with glazed and polished cement surfaces after thermal cycling in a storage media [lactic acid solution (pH 4.0)]. The changes were measured with regard to the surface roughness, colour-change, gloss, hardness, toothbrush wear, and SEM (scanning electron microscope) observation in the laboratory. The glazed cement surface was not affected after thermal cycling in the lactic acid solution. PMID- 7490669 TI - In vitro investigation of various attachments for overdentures on osseointegrated implants. AB - Primary splinting of fixtures with bar attachments has proved to be clinically effective for overdentures on osseointegrated implants in the edentulous mandible. However, in vitro investigations indicate that a more favourable force transfer to the individual fixtures can be attained by secondary splinting of four implants with conical double crowns. This observation could be confirmed in the present study by measurement of implant deflection in a model of the mandible by means of interferometric optical holography. Telescope crowns with conical interface brought about functionally more even movement characteristics of the implants compared to rigid or jointed bar attachments. Moreover, the conical double crowns always ensured a stable position of the denture in contrast to jointed attachments. These results underscore the need to test conical double crowns clinically as anchors for overdentures on osseointegrated implants. PMID- 7490670 TI - Asymmetry of masticatory muscle activity during intercuspal maximal clenching in healthy subjects and subjects with stomatognathic dysfunction syndrome. AB - The present study investigated the asymmetry of masticatory muscle activity during maximal intercuspal clenching in healthy subjects and subjects with stomatognathic dysfunction syndrome. Stomatognathic dysfunction syndrome, unilateral mastication and the asymmetry of masticatory muscle activity appear to be related to each other. The asymmetry of masseter muscle activity was greater as stomatognathic dysfunction became increasingly severe. Because fatigue and pain are produced more quickly by unilateral clenching than bilateral clenching, clenching under conditions of left and right muscular imbalance can further aggravate stomatognathic dysfunction. It is suggested the asymmetry of masseter muscle activity during maximal clenching correlates with the onset of the stomatognathic dysfunction syndrome. Stomatognathic dysfunction syndrome is closely related to the asymmetry of masseter muscle activity and only slightly related to the asymmetry of temporal muscle activity. The asymmetry of anterior temporal muscle activity appears to have little clinical significance. PMID- 7490671 TI - Stress analysis of four-unit fixed bridges on abutment teeth with reduced periodontal support. AB - The variable properties of photoelastic models and the difficulty of fabrication of the model limit experimental designs. The 'material fringe value' is introduced into the definition of the 'stress concentration index' by multiplying the fringe orders. Three levels of periodontal support in three photoelastic models (nine cases) were compared quantitatively by determining the stress concentration induced in periodontium by point loading vertically on a span of the fixed partial denture of four units length. The results indicate that comparison among photoelastic models is possible and useful. The stress concentration effects produced by the second molar were more prominent and determinant than that by the first premolar in the periodontium of the abutments. PMID- 7490672 TI - The effect of matrix phase morphology on the structure of Ag-Cu-Pd dispersed phase dental amalgam. AB - The effect of an Ag-Cu-Pd dispersant alloy on the structure of amalgams fabricated with two different particle types of low-copper amalgam matrices was investigated. In amalgams L and L (0), a low-copper lathe-cut amalgam Aristalloy was used as the matrix. In amalgams S and S(0), the matrix used was Spheralloy, a low-copper spherical amalgam. X-ray diffraction was used to analyse the relative content of the phases in the amalgams. The presence of Pd in the dispersant of amalgams L and S resulted in a decrease in the amount of eta' (Cu6Sn5) and an increase in the amount of unreacted particles when compared to the controls L(0) and S(0). Amalgams fabricated with a spherical alloy matrix (S and S(0)) showed a greater effect than amalgams fabricated with a lathe-cut matrix alloy (L and L(0)). PMID- 7490673 TI - Comparative studies on check-bite and central-bearing-point method for the remounting of complete dentures. AB - Thirty-six complete dentures were remounted both with check-bite as well as central-bearing-point (CBP) registration. The mandibular or denture position was registered in the condylar area three times with each method. Two independent operators carried out the registrations. One half of patients had their occlusion equilibrated according to the check-bite registration, the other half in the central-bearing-point position. After individual determination of the hinge axis the upper dentures were mounted by a face-bow transfer into a Dentatus articulator. A total of six registrations was performed and repeatedly compared in the condylar area in three dimensions using a computer supported measuring device. The reproducibility of the check-bite registration was 0.37 +/- 0.33 mm, with a maximum of 1.77 mm. For the central-bearing-point method this was 0.29 +/- 0.26 mm with a maximum of 1.56 mm. The mean difference in condylar position between both methods was 0.66 +/- 0.55 mm with a maximum of 4.02 mm. A third independent operator re-examined 30 of the patients 2-3 weeks after the remounting and evaluated the adaptation to the new dentures. Those patients whose dentures were equilibrated with the central-bearing-point method tended to cope better with their dentures and suffered on average from fewer pressure spots. PMID- 7490674 TI - The effect of the slope inclination of buccal polished surface of mandibular denture base on food retention. AB - The accumulation of food debris in the buccal vestibule can inhibit normal mastication and appears to be a factor that can cause patient discomfort. This study focused on the effect of the slope inclination of the buccal polished surface of the mandibular denture base on the ability to expel test-food particles from the buccal vestibule. It was revealed that an inclination of the slope steeper than 80 degrees markedly inhibits the normal ability to expel foods from the vestibule. It was revealed that the most appropriate inclination of the buccal slope is approximately 60 degrees or 70 degrees. These results could be utilized for constructing a denture prosthesis. PMID- 7490675 TI - Intestinal lymphoma and enteropathy. PMID- 7490676 TI - Non-Hodgkin's lymphoma, coeliac disease, and Epstein-Barr virus: a study of 13 cases of enteropathy-associated T- and B-cell lymphoma. AB - A group of 166 patients with coeliac disease was followed for a period of up to 25 years. During this time, 17 patients developed intestinal tumours that were diagnosed as lymphoma, of which 15 cases were available for review. Eleven of the lymphomas were of T-cell type (enteropathy-associated T-cell lymphoma, EATL) and two were of B-cell type. Two cases were reclassified as undifferentiated carcinoma. The interval between the diagnosis of enteropathy and the onset of lymphoma varied from less than 2 months in four patients to more than 5 years in seven. Seven of the T-cell and both B-cell lymphomas were investigated for the presence of Epstein-Barr virus (EBV) by in situ hybridization (ISH) using probes against Epstein-Barr virus-encoded RNAs (EBERs) and by immunohistochemistry with EBV-specific monoclonal antibodies. All EATL cases were negative, suggesting that EBV is not an important factor in these cases. In one of the B-cell cases, EBV was detectable by ISH and immunohistochemistry in most tumour cells in the mesenteric lymph nodes, but not in any of the tumour cells in the primary ileal tumour, indicating that in this case EBV infection was a late event in the neoplastic process. These results show that lymphoma may develop any time after the onset of coeliac disease and that in our cases of EATL, EBV was not an important factor. In some cases of EBV-related neoplasia, virus infection may be a late event. PMID- 7490677 TI - Determination of transforming growth factor beta 1 mRNA expression in breast carcinomas by in situ hybridization. AB - The expression of transforming growth factor beta 1 (TGF-beta 1) mRNA has been determined in 16 breast carcinomas using in situ hybridization and compared with TGF-beta protein as detected by antibodies against TGF-beta 1 and TGF-beta 1 plus TGF-beta 2. Digoxigenin-labelled riboprobes were used, with alkaline phosphatase and immunogold silver detection systems. TGF-beta 1 mRNA was only detected in carcinomas in which TGF-beta 1 protein was found (9 of 16 cases) and not in those with prominent reactivity for TGF-beta 2. RNA preservation was poor in two other cases in which TGF-beta 1 protein had been detected. In general, those tumours with greater numbers of cells labelled for TGF-beta 1 mRNA had prominent reactivity for TGF-beta 1 protein. The mRNA was localized to cancer cells with no labelling of stromal cells, although in a small number of cases scanty staining for TGF-beta 1 protein had been observed in stromal cells. The incidence of detection of TGF-beta 1 mRNA is lower than the published data from Northern analysis studies of breast carcinomas, suggesting that only higher levels of TGF beta 1 mRNA expression are being detected by in situ hybridization. However, this approach has provided useful information about the cellular sites of expression of TGF-beta 1 in breast carcinomas. PMID- 7490678 TI - Detection of loss of heterozygosity of p53 gene in paraffin-embedded breast cancers by non-isotopic PCR-SSCP. AB - A rapid non-isotopic PCR-SSCP (polymerase chain reaction-single-stranded conformation polymorphism) method was developed in this study to detect polymorphism and loss of heterozygosity (LOH) of p53 in formalin-fixed and paraffin-embedded samples of normal breast tissue and of breast cancer. p53 expression was also examined by immunohistochemistry. In 35 paired samples, heterozygosity in exon 4 of p53 was detected in 17 cases (49 per cent) and LOH of the p53 gene in breast cancer tissues was observed in 7 out of 15 informative cases (47 per cent). The correlation of LOH of p53 with positive p53 immunostaining did not reach statistical significance, but all immunostaining positive tumours among informative cases had LOH of p53. The results support the hypothesis that in most cases the allelic deletion of p53 may uncover the 'recessive mutation' in the remaining allele. However, LOH of p53 was more frequent than positive immunostaining and was significantly associated with poor differentiation of breast cancer (P < 0.05). The results suggest that the allelic deletion of p53 may also contribute to the development and progression of breast cancer by reducing the amount of normal p53 protein. These results show that non isotopic PCR-SSCP is a simple, fast, and effective method for detecting polymorphism and LOH of the p53 gene, which is especially useful for retrospective studies. PMID- 7490679 TI - Expression of bcl-2 oncogene protein is prevalent in small cell lung carcinomas. AB - To investigate the frequency of bcl-2 oncogene protein expression in small cell lung carcinoma (SCLC), immunohistochemical staining with a mouse-anti-human monoclonal antibody, bcl-2/124, was carried out on 60 formalin-fixed, paraffin embedded SCLC samples obtained from surgical biopsy, and autopsy cases. bcl-2 protein was detected in 54 out of the 60 SCLCs. In 47 cases, more than half of the tumour cells stained positively. The staining intensity of the tumour cells was comparable to that of infiltrating lymphocytes in 37 cases, but varied from area to area and even from cell to cell. Negative data in six cases were found to be due to unsuitable fixation or embedding procedures rather than the absence of the antigen. bcl-2 oncogene protein may thus be expressed in most if not all SCLCs. bcl-2 may have potential diagnostic and therapeutic importance in SCLCs and non-SCLCs. Previous cytogenetic and molecular genetic analyses indicate that SCLCs carry a number of chromosomal abnormalities and it would follow from the present results that the abnormal expression of bcl-2 may also play a role in the pathogenesis of SCLC, by increasing tumour mass through inhibition of apoptosis as previously proposed. The diagnostic, prognostic, and therapeutic implications of these findings should be studied in greater detail. PMID- 7490680 TI - Expression of the c-kit gene product in normal and neoplastic mast cells but not in neoplastic basophil/mast cell precursors from chronic myelogenous leukaemia. AB - The expression of the c-kit gene product has been examined in normal mast cells, mast cell neoplasms, and basophil/mast cell precursors obtained from patients with chronic myelogenous leukaemia (CML). Formalin-fixed, paraffin-embedded sections or smears fixed with formalin vapour were studied by immunohistochemical methods, using a polyclonal antibody against the c-kit gene product. Normal and neoplastic mast cells showed a positive immunoreaction for c-kit gene product, but neoplastic basophil/mast cell precursors from CML patients lacked c-kit gene product by immunohistochemical and flow cytometric methods, even in cells having mast cell granules, together with or without basophil granules. Mast cell tryptase was, however, expressed in normal and neoplastic mast cells and basophil/mast cell precursors containing mast cell granules. In addition, cells of monocyte/macrophage lineage lacked c-kit gene product. These findings indicate that the c-kit gene product may play an important role in the development and function of mast cell but not of cell of basophil and monocyte/macrophage lineage. PMID- 7490682 TI - Prostatic intraepithelial neoplasia (PIN). Performance of Bayesian belief network for diagnosis and grading. AB - Prostatic intraepithelial neoplasia (PIN) diagnosis and grading are affected by uncertainties which arise from the fact that almost all knowledge of PIN histopathology is expressed in concepts, descriptive linguistic terms, and words. A Bayesian belief network (BBN) was therefore used to reduce the problem of uncertainty in diagnostic clue assessment, while still considering the dependences between elements in the reasoning sequence. A shallow network was used with an open-tree topology, with eight first-level descendant nodes for the diagnostic clues (evidence nodes), each independently linked by a conditional probability matrix to a root node containing the diagnostic alternatives (decision node). One of the evidence nodes was based on the tissue architecture and the others were based on cell features. The system was designed to be interactive, in that the histopathologist entered evidence into the network in the form of likelihood ratios for outcomes at each evidence node. The efficiency of the network was tested on a series of 110 prostate specimens, subdivided as follows: 22 cases of non-neoplastic prostate or benign prostatic tissue (NP), 22 PINs of low grade (PINlow), 22 PINs of high grade (PINhigh), 22 prostatic adenocarcinomas with cribriform pattern (PACcri), and 22 prostatic adenocarcinomas with large acinar pattern (PAClgac). The results obtained in the benign and malignant categories showed that the belief for the diagnostic alternatives is very high, the values being in general more than 0.8 and often close to 1.0. When considering the PIN lesions, the network classified and graded most of the cases with high certainty. However, there were some cases which showed values less than 0.8 (13 cases out of 44), thus indicating that there are situations in which the feature changes are intermediate between contiguous categories or grades. Discrepancy between morphological grading and the BBN results was observed in four out of 44 PIN cases: one PINlow was classified as PINhigh and three PINhigh were classified as PINlow. In conclusion, the network can grade PIN lesions and differentiate them from other prostate lesions with certainty. In particular, it offers a descriptive classifier which is readily implemented and which allows the use of linguistic, fuzzy variables. PMID- 7490681 TI - The immunohistochemical localization of drug-metabolizing enzymes in prostate cancer. AB - The major groups of enzymes involved in activating and detoxifying therapeutic drugs, not least several anti-cancer drugs, include the cytochromes P450 (P450s), epoxide hydrolase, and glutathione S-transferases (GSTs). The expression of these enzymes in malignant tumours is one possible mechanism of anti-cancer drug resistance. This study has investigated the presence, cellular localization, and distribution of drug-metabolizing enzymes in prostate cancer. The P450 subfamilies CYP1A, CYP2C, and CYP3A were present in 63, 25, and 61 per cent of tumours, respectively. Epoxide hydrolase was identified in 96 per cent of tumours. GST-alpha and GST-mu were expressed in 29 and 41 per cent of tumours, respectively, while there was no immunoreactivity for the pi form of GST. The absence of GST-pi in prostate cancer contrasts with the frequent expression of GST-pi observed in other types of malignant tumour. In non-neoplastic prostatic epithelium, there was expression of CYP1A, CYP2C, epoxide hydrolase, and the different forms of GST, while there was no apparent immunoreactivity for CYP3A. PMID- 7490683 TI - Immunohistochemical localization of ubiquitin cross-reactive protein in human tissues. AB - Ubiquitin cross-reactive protein (UCRP) is an interferon-inducible ubiquitin homologue which is constitutively present in cells and can be conjugated to other proteins. Using a characterized polyclonal antiserum to UCRP, immunohistochemical localization of UCRP was performed on paraffin-processed normal human tissues and in human tissues known to contain ubiquitinated intracellular inclusions. The antibody to UCRP immunostained lymphoid cells, striated and smooth muscle, several epithelia, and neurons. The level of staining varied greatly between tissues but was in a consistent punctate pattern. Localization to neuromuscular junctions and striations is similar to that described for antisera to ubiquitin protein conjugates. Inclusion bodies characterized by immunoreactivity to anti ubiquitin were not detected by the antibody to UCRP. Importantly, because UCRP may also be detected by antisera to conjugated ubiquitin, future studies on the distribution of ubiquitin in tissue sections must now take account of possible cross-reactivity with UCRP. PMID- 7490684 TI - The intracellular polymerase chain reaction for small CMV genomic sequences within heavily infected cellular sections. AB - The indirect intracellular polymerase chain reaction (in situ PCR) combines the potential sensitivity of the polymerase chain reaction (PCR) with the high specificity and morphological preservation of in situ hybridization (ISH). This study describes a method for the amplification of small, specific cytomegalovirus (CMV) genomic sequences [100 base pairs (bp)] within large formalin-fixed, paraffin-embedded tissue sections. A heat-resistant glue surrounds the section, creating a well which contains a relatively large volume of isotonic reaction solution without a covering mineral oil layer; this optimizes morphological preservation, permits the evaluation of large sections, and allows both denaturation steps and up to 40 cycles of in situ PCR to be performed, whilst progressively concentrating the reaction solution by evaporation during thermal cycling. ISH was performed using a non-isotopic DNA probe with specificity for CMV, with or without prior in situ PCR amplification, both for samples on slides (fibroblasts and lung) and in suspension (fibroblasts). Samples on slides were evaluated by both blinded studies and image analysis, comparing the intensity of signal (P < 0.003) and the numbers of positive cells detected (P < 0.007), with or without intracellular amplification. Cells in suspension were analysed by blinded studies on cytospins and by gel electrophoresis of cell lysates. Successful intracellular amplification was achieved in this high copy model. PMID- 7490685 TI - A megakaryocyte analysis of the bone marrow in patients with myelodysplastic syndrome, myeloproliferative disorder and allied disorders. AB - A morphometric analysis was performed on aspirate clots of bone marrow to identify the presence of atypical megakaryocytes after immunohistological staining with a monoclonal antibody against Factor VIII. This study included cases of myelodysplastic syndrome (MDS), myeloproliferative disorder (MPD), aplastic anaemia (AA), idiopathic thrombocytopenic purpura (ITP), chronic myelogenous leukaemia (CML), and control cases free from any haematological disease. Quantitative and qualitative abnormalities of megakaryocytes were assessed using an image analyser and a personal computer to perform a morphometric analysis of the number (per mm2), arrangement (microns), nuclear size (microns2), cell size (microns2), nuclear size/cell size (N/C) ratio, and nuclear contour index (NCI). Micromononuclear megakaryocytes were detected in MDS, while in MPD, large over-mature magakaryocytes were observed to increase in number. In AA, the megakaryocytes decreased dramatically in number without showing any morphological abnormality. In CML, the megakaryocytes also increased without any remarkable morphological abnormality. In ITP, only the NCI increased. The above results show that MDS is characterized by the presence of atypical micromegakaryocytes, while MPD is characterized by atypical large over-mature megakaryocytes. PMID- 7490686 TI - Pathogenesis of segmental glomerular changes at the tubular origin, as in the glomerular tip lesion. AB - Segmental abnormalities at the glomerulo-tubular junction, or tip changes, are a common and early feature in several experimental and human renal disorders. Scanning and transmission electron microscopic study of an experimental model, Lewis rats given anti-glomerular basement membrane antibodies, showed that at 5 days there were monocytes/macrophages in glomerular capillary loops next to the tubular origin, with abnormalities of visceral epithelium, including prolapse of groups of these cells into the tubular origin. At 12 days, there was contact and adhesion between glomerular capillary loops and Bowman's capsule. Transmission electron microscopic study of human renal biopsies showed that the human tip changes resembled the later lesions seen in rats, with intracapillary foam cells confirmed by immunohistological study to be monocytes/macrophages. These findings show that abnormalities of the glomerular tuft precede adhesion to Bowman's capsule and that the earliest changes recognized by light microscopy in human renal biopsies are relatively late in their natural history, meaning at least several days old. Prolapse of visceral epithelial cells into the tubular origin may explain the characteristic site of tip changes. PMID- 7490687 TI - Spatial arrangement of IgA and C3 as a prognostic indicator of IgA nephropathy. AB - The histological picture of primary glomerulonephritis with glomerular IgA deposition (IgA nephropathy and Henoch-Schonlein disease) can vary from minimal mesangial involvement to severe endocapillary and/or extracapillary proliferation. Local activation of the complement cascade by glomerular IgA deposits and release of anaphylactoid factor are considered to be major triggers of inflammation, but clear-cut correlations between the severity of the histological findings and the intensity of glomerular deposition of immunoglobulin and complement fractions are still lacking. The purpose of this study was to investigate the spatial distribution of IgA and complement in mesangial deposits with confocal laser scanning microscopy (CLSM) and to correlate specific patterns of IgA-complement interaction with glomerular damage. Two groups of patients have been studied, one with mild to moderate diffuse mesangial proliferation and the other with diffuse proliferative endocapillary and/or extracapillary patterns. In milder forms of the disease, the majority of the immune deposits are composed of both IgA and C3, coated by an outer layer of IgA alone. Large C3 deposits, or deposits composed of IgA and C3 without an outer coat of IgA, were associated with more severe histological lesions. The results suggest that free access of active complement components to cell and/or mesangial matrix receptors could trigger a cytolytic reaction and that immunoglobulins seem to act as a protective layer on C3 components. PMID- 7490688 TI - Human malignant mesothelial cells: variability of ultrastructural features in established and nude mice transplanted cell lines. AB - The aim of this study was to determine, by transmission electron microscopy, the differentiation features of 21 human malignant mesothelioma cell lines (HMCLs) established from 13 specimens of 12 confirmed human malignant mesotheliomas, and of tumours induced in nude mice injected with 16 HMCLs. Fifty per cent of HMCLs showed typical mesothelial differentiation (long and slender microvilli, desmosomes, perinuclear intermediate filaments); 29 per cent did not show differentiation; and the remainder were poorly differentiated. Three human tumour specimens gave several different HMCLs; the cell lines obtained from a given tumour exhibited variable mesothelial differentiation. Eleven HMCLs were compared with the native tumour. Four were similar to the tumour and seven were less well differentiated, in most cases in relation to their microvilli. With six HMCLs, tumours induced in nude mice were less well differentiated than the corresponding cell lines, whereas with four HMCLs, tumours were equally or better differentiated. However, in most nude mice tumours, typical mesothelial microvilli were present. These results show that cell lines established from malignant mesothelioma may exhibit dedifferentiated features. However, while the variability in ultrastructural differentiation may result from the culture microenvironment, it could also be related to the state of differentiation, of the native tumour sample and to tumour cell heterogeneity. PMID- 7490689 TI - Cardiac rehabilitation: how can it be improved? PMID- 7490690 TI - Vascular risk factors and atherosclerosis as risk factors for cognitive decline and dementia. PMID- 7490691 TI - Organic and psychosocial risk factors for duodenal ulcer. AB - Duodenal ulcer (DU) is a common medical disorder which frequently becomes chronic. The factors involved in the aetiology of DU are poorly understood. Evidence for the contribution of psychosocial factors to the onset of DU is reviewed in the context of evidence of organic factors, genetic and environmental, with special reference to new evidence of infectious agents in the aetiology of DU. Recent progress in understanding of biological forces of duodenal ulcer disease has lead to a substantially revised view of psychosomatic factors. Although these may be of significance, their relative contribution to aetiology is likely to be modest. PMID- 7490692 TI - Cholesterol and psychological well-being. AB - The debate about possible adverse effects associated with low or lowered serum cholesterol has raised important scientific questions concerning the links between lipids and behaviour. One of the most unexpected findings has been an association between cholesterol-lowering treatment and accidental death. A similar association has also emerged among the prospective cohort studies, with higher-than-expected numbers of suicide deaths in the lowest cholesterol groups. These observations have prompted speculation that behavioural or emotional disturbances could be part of the process linking lipids and accidental death. In this paper, the epidemiological literature is reviewed briefly, then the evidence for depression as a mediating condition is discussed. Two conclusions are drawn from this review of the literature. One is that understanding the relationship between the biology of lipids and the psychobiology of mood is demonstrably an important scientific and public health issue. The second is that the introduction of new treatments or preventive programmes should include a careful evaluation of the psychological as well as the physical effects. PMID- 7490693 TI - Psychosocial factors and pregnancy outcome: a review with emphasis on methodological issues. AB - This review focuses on the research concerning the relation between psychosocial factors and pregnancy outcome. The following four outcome measures are dealt with: (1) birth weight, (2) preeclampsia, (3) preterm labour, and (4) intrapartum complications. The most consistent finding concerns the association between maternal exposure to taxing situations and preterm delivery. Three possible pathways are hypothesized: (1) an indirect influence via unhealthy coping and life style behaviour, (2) a direct influence via stress-dependent hormones, and (3) an additional direct influence via psycho-immunological factors. Intervention studies aimed at improving pregnancy outcome show fairly mixed results. It is recommended that studies on the relationship between psychosocial factors and pregnancy outcome should employ a prospective design with due attention to chronic stressors, should include appropriate biochemical assessments, and multivariate techniques are applied. PMID- 7490694 TI - A controlled study of anxiety and morbid cognitions at initial screening for human immunodeficiency virus (HIV) in a cohort of people with haemophilia. AB - AIM: This study examines the relationship between anxiety, psychological state and Human Immunodeficiency Virus (HIV) stages as defined by the Centers for Disease Control at the time of initial screening for HIV in a cohort of people with haemophilia who were at risk of prior exposure to HIV transmission from blood products. METHOD: Psychological scores, immunological measures, and clinical data from case notes for 116 potentially HIV exposed people with haemophilia attending initial screening for HIV infection in 1984-1985, were used to examine the relationship between psychological variables, clinical state and their clinical classification under the Centres for Disease Control categorization. Psychometric test results were obtained for 63 HIV seronegative patients and 53 HIV seropositive patients. Planned comparisons, multiple and logistic regressions, were used to explain observed differences between seronegative and seropositive subjects. The potential confounders of sex, age, severity of haemophilia, haemophilia type and blood product usage were controlled. RESULTS: The major finding of this study was that higher levels of State Anxiety at the time of initial screening for HIV, were observed in those patients who lacked recognized symptoms of HIV infection and were seropositive, compared with seronegative subjects. The State Anxiety scores were predicted by HIV infection or alternatively CD4+ T-cell levels. CONCLUSION: The findings of this study suggest that HIV infection can produce psychological effects prior to any physical symptoms of infection being apparent. PMID- 7490695 TI - Distribution of positive moods in women with the premenstrual syndrome and in normal women. AB - A daily mood record was kept by 44 women presenting with PMS (PMS+ group: 133 menstrual cycles) and by 48 normal women (PMS- group: 100 cycles). Peaks of maximum positive mood were located after fitting either a 5-term or a 3-term Fourier series to the data. In the PMS+ group mood peaks were clustered to a significant degree (p < 0.001) around day 11 in a 28-day menstrual cycle (95% confidence interval: days 10-12); this is the time of the pre-ovulatory oestrogen surge. In the PMS- group clustering was insignificant. Menstrual cycle parameters were similar in the two groups (PMS+ vs. PMS-: mean duration cycle, 27.6 +/- 3.2(SD) vs. 26.7 +/- 2.5 days; incidence ovulatory cycles, 95.5 vs. 93.0%). The evidence suggests either that the ovarian hormone cycle has no effect on mood, or that it has an effect only in women with PMS. PMID- 7490697 TI - The risk of seclusion and the menstrual cycle in female psychiatric patients. AB - The alleged clinically observed association between the use of seclusion and the luteal phase of the menstrual cycle was studied in 113 consecutively admitted female psychiatric patients by means of a monitoring of oestrogen and progesterone levels, the use of seclusion, age, drug use, regularity of menses, and a history of hysterectomy. Twenty-one patients (18.6%) were in the luteal phase. During the study period, 14 patients (12.4%) had to be secluded. No significant difference in the frequency of seclusion between luteal patients and patients in the other phases could be demonstrated. The findings indicate that the luteal phase of the menstrual cycle is not associated with an increased occurrence of seclusion in female psychiatric patients. PMID- 7490696 TI - Causal attribution and adaptation among lung cancer patients. AB - The aim of the present study was to describe lung cancer patients' causal attributions and examine their associations with adaptation. Methods were based on semi-structured interview, content analysis, self-reports, interviewer ratings and standardized questionnaires. 'Smoking cigarettes' and 'toxins in the work place' were the most commonly mentioned possible causes. Patients who made a psychosocial causal attribution suffered greater emotional distress, were more depressed, and less hopeful than other patients. They were also more likely to be rated as showing a maladaptive way of coping with illness. The implications of these findings for psychosocial care are discussed. PMID- 7490698 TI - Does high 'action-proneness' make people more vulnerable to chronic fatigue syndrome? A controlled psychometric study. AB - Degree of premorbid 'action-proneness' was measured, using a self-administered questionnaire, in 35 patients suffering from chronic fatigue syndrome (CFS), all the members of 'ME'-self help groups and all those meeting CDC-criteria of CFS. The results were compared with those of 30 chronic idiopathic musculoskeletal pain patients, 34 patients with a chronic organic condition, and 34 neurotic patients without primary somatic complaints. Statistical analysis showed that CFS patients described themselves as significantly more 'action-prone' than the last two groups, and to a degree which was comparable with the chronic pain group. The results could not be explained by concomitant depression and are in accordance with anecdotal reports of premorbid hyperactive lifestyle in CFS patients. Further investigations seem worthwhile to test the hypothesis that hyperactivity might be a predisposing factor for chronic illness behaviour in CFS patients. PMID- 7490699 TI - Routine stresses in caring for a child with cystic fibrosis. AB - Difficulties experienced by mothers caring for a child with cystic fibrosis were assessed. Difficulties were categorized as: (1) developmental, which included tasks generic to all families (bedtimes, socialization, siblings and communication with others); and (2) illness, which were especially pertinent to the care of the child with cystic fibrosis (mealtimes, physiotherapy and home based medical care). Age differences were found in stresses associated with developmental routines, with mothers of younger children reporting more difficulties. With regard to illness routines, more difficulties were reported for children in better health (higher Shwachman scores). The data suggest that parents experience increasing difficulties in implementing home-based medical care where the child is in better health. Clinic staff need to be more aware of these difficulties and target information to parents appropriately. PMID- 7490700 TI - Prediction of the developmental potential of hamster embryos in vitro by precise timing of the third cell cycle. AB - Time-lapse videomicrography was used to determine the timing of early developmental events in hamster embryos in vitro. The time intervals from pronuclear envelope breakdown to the completion of the first cleavage (Dt2), second cleavage (Dt4 = 2-4 cells), third cleavage (Dt8 = 4-8 cells), blastocyst formation, and zona escape were precisely measured to determine whether the variable 'time' (t) can be used to predict the developmental potential of preimplantation embryos. The range of the developmental time interval (Dt) from the second to the third cleavage divisions (Dt8) provided the best indicator for predicting the probabilities of blastocyst formation and zona escape (P = 0.015 and 0.041, respectively). Dt8 was subdivided into consecutive time cutoff points of < or = 750, < or = 800, < or = 850 and < or = 900 min. Of the embryos that took < or = 750 min to complete the third cleavage division, 92% developed into blastocysts and 69% escaped from their zonae pellucidae. When the completion of Dt8 extended to < or = 900 min, the percentages decreased to 75% and 49% for blastocyst formation and zona escape, respectively. This study identifies a specific developmental time interval and a model whereby time can be used as a noninvasive parameter to predict embryo developmental potential in vitro. PMID- 7490701 TI - Stimulation of motility and respiration of intact fowl spermatozoa by calyculin A, a specific inhibitor of protein phosphatase-1 and -2A, via a Ca2+-dependent mechanism. AB - Intact fowl spermatozoa became almost immotile at 40 degrees C. In contrast, the presence of 10-1000 nmol calyculin A l-1, a specific inhibitor of protein phosphatase-1 (PP1) and -2A (PP2A), permitted activation of sperm motility in a dose-dependent manner. Calyculin A also stimulated the rate of oxygen consumption by spermatozoa, and induced a concomitant decrease in ATP concentrations, suggesting a coupling of ATP hydrolysis to the rate of oxidative phosphorylation. However, the motility and oxygen consumption of spermatozoa loaded with an intracellular Ca2+ chelator, 1,2-bis (2-aminophenoxy)ethane-N,N,N',N'-tetraacetic acid, tetraacetoxymethyl ester (BAPTA/AM), were not stimulated by calyculin A alone, but only after the subsequent addition of 2 mmol CaCl2 l-1. These results suggest that inhibition of the activities of endogenous PP1 and PP2A may stimulate the motility and metabolic activity of fowl spermatozoa at 40 degrees C via a mechanism that requires intracellular free Ca2+. PMID- 7490704 TI - Effects of a second insemination after ovulation on fertilization rate and accessory sperm count in sows. AB - Insemination immediately after ovulation causes low fertilization results owing to a low fertilization rate and possibly also owing to polyspermic fertilization. The present experiment was undertaken to study the effects of a second insemination after ovulation on fertilization rate and embryonic development. In multiparous crossbred sows, transrectal ultrasonography was used at intervals of 4 h to determine ovulation. All sows (n = 91) were artificially inseminated with 3 x 10(9) mixed spermatozoa and, in 31 sows, a second insemination took place at 3 +/- 1 (mean +/- SD) h after ovulation. At 119 +/- 5 h after ovulation, the percentage of normal embryos and the accessory sperm count were determined. In the sows that were inseminated once, the percentage of normal embryos decreased when insemination took place more than 24 h before ovulation, from 88 +/- 20% (16 24 h; n = 15) to 63 +/- 40% (24-32 h; n = 10) (P < 0.05). In the sows that were inseminated again after ovulation, the percentage of normal embryos was high, irrespective of the period between first insemination and ovulation. The difference in percentage of normal embryos between the sows that were inseminated once or twice was significant when the first insemination took place between 24 and 32 h before ovulation; 63 +/- 40% (inseminated once, n = 10) and 97 +/- 5% (inseminated twice, n = 8) (P < 0.05). The accessory sperm count of the normal embryos in a litter was positively related to the percentage of normal embryos in a litter, and this relation was not affected by the interval between first insemination and ovulation or by the number of inseminations.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7490702 TI - Stimulation of endocytosis in mouse blastocysts by insulin: a quantitative morphological analysis. AB - The effects of insulin on the endocytic activity of mouse blastocysts in vitro were investigated using confocal laser scanning microscopy, quantitative image analysis and electron microscopy. Confocal studies showed that fluorescein isothiocyanate-labelled markers, dextran (fluid phase) and albumin (combined membrane and fluid phase), were endocytosed by blastocysts and localized within vesicles (about 2.5 microns in diameter) in the outer trophectoderm cells. No labelling was detected in the inner cell mass cells or the blastocoel cavity. Treatment with 170 nmol insulin l-1 stimulated the endocytosis of fluorescently labelled dextran in freshly collected blastocysts, increasing mean vesicle diameter per embryo by 15% (P < 0.05) after incubation with insulin for 2.5 h and mean vesicle number per embryo by 56% (P < 0.01) after 6 h. Both effects were also evident in blastocysts that had been cultured from the late eight-cell stage. Blastocysts incubated for 6 h with insulin displayed increased convolutions in the trophectoderm apical membrane compared with controls, indicating increased membrane activity and suggesting macropinosome formation. Collectively, these results suggest that insulin enhances endocytosis in the trophectoderm by stimulating uptake at the apical membrane into larger and more numerous endocytic vesicles and with some evidence of vesicle fusion. This mechanism may provide a metabolic basis for the stimulation by insulin of biosynthesis, proliferation and morphological development in early embryos. PMID- 7490703 TI - Stimulation of rat granulosa cell progesterone production but not other differentiated functions by a splenocyte-derived factor. AB - Concanavalin A-stimulated splenocytes secrete a factor that stimulates progesterone production in cultured rat granulosa cells. The actions of this progesterone-stimulating factor (PSF) were characterized further by purifying it by sequential chromatographies on heparin-agarose, copper chelating-Sepharose and Mono S columns. Several of its effects on granulosa cells were then compared with those of FSH. Like FSH, PSF induced accumulation of progesterone and 20 alpha dihydroprogesterone in granulosa cell culture media; however, the time course of progesterone accumulation in response to PSF was much slower than that in response to FSH. In contrast to FSH, PSF induced little accumulation of oestrogen. Induction of other differentiated responses was compared by pretreating cultured granulosa cells with either FSH or PSF, and then challenging with LH, prolactin, epidermal growth factor or the beta-adrenergic agonist, isoproterenol. Pretreatment with FSH induced subsequent responsiveness to all four agents, whereas pretreatment with PSF induced subsequent responsiveness only to isoproterenol. These results indicate that granulosa cell responses to PSF are much more limited than are those to FSH and that these responses are characterized by an increase in progestin production. PMID- 7490705 TI - Adenylyl cyclase activity in human decidua and myometrium during pregnancy and labour. AB - Adenylyl cyclase activity was studied in human decidua and myometrium in early pregnancy and at term before and after the onset of labour. Decidual basal, prostaglandin-, catecholamine- and forskolin-stimulated adenylyl cyclase activities at term before the onset of labour were significantly lower than those in early pregnancy. After the onset of labour at term, decidual basal, prostaglandin-, catecholamine-, NaF- and forskolin-stimulated adenylyl cyclase activities significantly increased compared with those at term before the onset of labour. Myometrial prostaglandin- and catecholamine-stimulated activities did not alter during pregnancy, except for basal and forskolin-stimulated activity. Myometrial basal, prostaglandin-, catecholamine-, NaF- and forskolin-stimulated activities at term showed no change after the onset of labour. At term, before the onset of labour, myometrial basal, prostaglandin, catecholamine-, NaF- and forskolin-stimulated activities were the same as those in the decidua. However, after the onset of labour at term, decidual basal and the stimulated activities were significantly higher than those in the myometrium. These results suggest that decidual prostaglandin- and catecholamine-stimulated adenylyl cyclase may play an important role in the initiation or maintenance of human labour or in both processes. PMID- 7490706 TI - Atretic changes of follicular wall caused by destruction of the germinal disc region of an immature preovulatory follicle in the chicken: an electron microscope study. AB - The germinal disc of the female gamete and its overlying follicular wall (the germinal disc region) is hypothesized to play an important role in the regulation of follicular growth. The role of the germinal disc region in follicular growth in chickens was investigated by destroying the germinal disc region or an area opposite the germinal disc (control) of the second largest follicle 22-23 h before ovulation of the F1 follicle, by localized freezing with solid CO2. Structural changes of the follicular wall (non-frozen region) were observed by electron microscopy 10-20 h after the destruction of the germinal disc region. Development of the inner structure of mitochondria in granulosa cells and accumulation of lipid droplets in thecal cells were observed in follicles 15 h after destruction of the germinal disc region. Twenty hours after destruction of the germinal disc region, follicles showed early signs of atresia (bursting atresia). Degenerative changes in follicles, including hydrolysis by lysosomal enzymes, were present in thecal fibroblast-like cells. Control follicles, in which an area opposite the germinal disc region was frozen 20 h before examination, had no degenerative features. These results provide further evidence that the germinal disc region is required for follicular growth. PMID- 7490707 TI - Menstrual cycle related differences in the proliferative responses of cultured human endometrial stromal cells to retinoic acid. AB - Although ovarian-derived steroid hormones are central to the control of endometrial growth and secretory differentiation, all-trans retinoic acid, a derivative of vitamin A, may also play an important role. Since the retinoids can inhibit the proliferation of both fibroblasts and epithelial cells in experimental systems, we investigated whether the proliferation of endometrial stromal cells was inhibited by all-trans retinoic acid. A sensitive spectrophotometric assay was used to show that the proliferation rates of primary cultures of endometrial stromal fibroblasts varied with the timing of biopsy, and that all-trans retinoic acid inhibited the growth of late secretory phase cells but had no effect on cells sampled at other times. Furthermore, since the expression of mRNA encoding cellular retinoic acid binding protein type II decreases in endometrial stromal cells in the secretory phase, a rise in intracellular all-trans retinoic acid concentrations could be fundamental to the control of endometrial stromal cell proliferation and differentiation in vivo. PMID- 7490708 TI - Endothelial cell death in preovulatory ovine follicles: possible implication in the biomechanics of rupture. AB - The follicular stigma that develops near the time of ovulation in sheep is void of thecal blood vessels. It was hypothesized that programmed apoptotic death of endothelial cells is a causative factor in this phenomenon. Apoptosis is a mode of physiological cell deletion that occurs during tissue regression and remodelling; it is characterized by oligonucleosomal fragmentation and cytoplasmic shrinkage. Direct fluorescence detection of digoxigenin end-labelled genomic DNA in situ was used as a marker of endothelial apoptosis in cross sections of the preovulatory ovine follicular wall. There was an abrupt increase in reactive cells after the gonadotrophin surge that was restricted to the presumptive ovulatory site immediately juxtaposed to the ovarian surface (the basal follicular endothelia remained unaffected). Vascular collapse and extravasation of blood cells preceded follicular rupture. Localized damage to thecal blood vessels and haemorrhagic tissue ischaemia may be contributing determinants of follicular instability and ovulation. PMID- 7490709 TI - Local action of trophoblast interferons in suppression of the development of oxytocin and oestradiol receptors in ovine endometrium. AB - Luteolysis in sheep is associated with uterine secretion of pulses of prostaglandin F2 alpha (PGF2 alpha) due to the action of luteal oxytocin on endometrial oxytocin receptors. For pregnancy to become established inhibition of oxytocin receptors is important as an antiluteolytic mechanism. The maternal recognition of pregnancy in cattle and sheep involves production, by the trophoblast, of a type 1 interferon (IFN-tau) that suppresses uterine development of oxytocin receptors and the generation of luteolytic episodes of PGF2 alpha. The action of IFN-tau in surgically prepared unilaterally pregnant ewes was investigated. Finn-Dorset ewes were anaesthetized on day 6 or 7 of the oestrous cycle and one uterine horn was surgically isolated at the uterine bifurcation from the body of the uterus. Ewes were mated at the subsequent oestrus either by a fertile or by a vasectomized ram and killed on day 13 or 16 after mating. On day 16, in the non-pregnant ewes, there was no measurable uterine IFN-tau but there were high concentrations of oxytocin receptors in both horns. In the pregnant ewes on day 16 after mating, the oxytocin receptor concentration was 45 +/- 11 fmol mg-1 protein in the pregnant horn and 585 +/- 131 fmol mg-1 in the non-pregnant horn. Antiviral activity was 5.8 x 10(7) +/- 5.2 x 10(7) U ml-1 in the pregnant horn and 2.9 x 10(3) +/- 1.2 x 10(3) U ml-1 in the non-pregnant horn. Thus, 16 days after mating, the pregnant horn exhibited high antiviral activity but oxytocin receptors were suppressed, while in the same endocrine environment (characteristic of pregnancy) there were low IFN-tau and high oxytocin receptor concentrations in the isolated horn equivalent to those expected at the onset of luteolysis. In situ hybridization to ovine mRNA encoding the oxytocin receptor and autoradiographic studies using the 125I-labelled oxytocin antagonist d(CH2)5[Tyr(Me)2,Thr4,Tyr-NH2(9)]-vasotocin both showed that the large amount of oxytocin receptor message and binding sites in the endometrium of the isolated horn were localized in the luminal epithelium. Immunocytochemical studies showed that there was a suppression of oestradiol receptors in the pregnant horn but high concentrations equivalent to those at oestrus were present in the isolated horn. The content of progesterone receptors was low in the stromal tissue only in both horns, a pattern of localization similar to that seen in the late luteal phase and in early pregnancy.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 7490710 TI - Detection of bovine viral diarrhoea virus antigen and RNA in oviduct and granulosa cells of persistently infected cattle. AB - Large-scale in vitro bovine embryo production systems commonly use genital tracts obtained from an abattoir as a source of both cumulus-oocyte complexes and co culture feeder cells. Tissues derived from this source may be contaminated with non-cytopathogenic bovine viral diarrhoea virus (BVDV) since, in several countries surveyed, approximately 1% of animals tested are persistently infected with this pathogen. Therefore, the use of such material in in vitro fertilization systems presents a potential risk for the transmission of BVDV to bovine embryos and via embryo transfer. This potential was investigated by obtaining oviduct epithelial cells and granulosa cells, which are commonly used as feeder cells, from cattle persistently infected with BVDV and examining them for the presence of BVD viral antigen (p80 non-structural protein and gp53 envelope glycoprotein) by indirect immunofluorescent histochemistry, and also viral RNA (encoding the p80 region) by in situ hybridization. In addition, titres of virus present in oviduct, ovary and blood were assayed by immunodetection on calf testis cell cultures. Luminal epithelial cells from the oviduct and primary cultures of granulosa cells and oviduct epithelial cells from such cattle were shown to contain both viral antigen and RNA. The susceptibility of both cell types to BVDV infection was further established by inoculating primary cell cultures of cells derived from cattle not infected with BVDV with a cloned isolate of non cytopathogenic BVDV (Pe515). RNA encoding BVDV and the antigen were detected 12 h after inoculation.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7490711 TI - Identification of male germ cells undergoing apoptosis in adult rats. AB - The possible role of apoptosis in spontaneous or induced germ cell death was investigated by treating adult male rats with either a GnRH antagonist (112.5 micrograms kg-1 day-1 for 14 days) or methoxyacetic acid (650 micrograms kg-1; single dose) or sham-treated with either of the vehicles (n = 3 per group). The antagonist virtually abolished gonadotrophin secretion, while methoxyacetic acid reduced serum testosterone concentrations and slightly increased those of FSH (neither significantly). Bands of low molecular mass characteristic of apoptotically degraded DNA were detected by electrophoresis in both treatment groups but not in the controls. Sectioned, Carnoy-fixed testes were screened for degenerating cells with periodic acid-Schiff's base and haemalaun or examined for apoptotic cells using a modified in situ end-labelling procedure. Periodic acid Schiff's-stained dying cells were found in low numbers in control animals with a distribution and frequency that matched that of apoptotic cells. Degenerating germ cells identified by histology were present at certain stages of spermatogenesis after 2 weeks of antagonist treatment. A comparison of their distribution with that of end-labelled cells identified the cell death as apoptotic. Methoxyacetic acid caused a massive depletion of spermatocytes at stages IX-II, which was also found to be apoptotic. It is concluded that spontaneous germ cell death in adult rats is apoptotic and that both gonadotrophin ablation and administration of methoxyacetic acid can cause apoptosis in the germ cells of adult male rats, but via different routes. PMID- 7490712 TI - Production of exogenous gonadotrophin-neutralizing immunoglobulins in cats after repeated eCG-hCG treatment and relevance for assisted reproduction in felids. AB - Combination regimens of equine chorionic gonadotrophin (eCG) and human chorionic gonadotrophin (hCG) are used in ovarian stimulation protocols for assisted reproduction of felids. In the present study, domestic cats exhibited decreased ovarian responsiveness after repeated administration of eCG-hCG and a possible immunological mechanism for this ovarian refractoriness was investigated. An ELISA was used to analyse sera from male, naive female and previously eCG-hCG stimulated (1x, 3x and 4x) female cats for the presence of immunoglobulins binding to eCG, hCG and pig FSH (pFSH). The sera of cats receiving multiple eCG hCG injections, at intervals of 44-50 days, displayed greater eCG and hCG-binding than did the sera of male, naive female or female cats stimulated once, and demonstrated variable affinity for pFSH. In preovulatory and postovulatory ovarian stimulation assays, mice injected with an eCG-antisera mixture had lower ovary masses than did mice injected with eCG-saline and fewer ovulated oocytes compared with mice treated with eCG-naive sera. Treatment of queens that were refractory to eCG-hCG with a pFSH-hCG regimen caused a rebound in development of ovarian follicles but not in oocyte maturity. These studies indicated that repeated treatment of domestic cats with eCG and hCG may cause an immunologically mediated refractoriness to ovarian stimulation. Although alternative gonadotrophin regimens may alleviate this refractoriness, a preferable strategy might be the avoidance of potential immunological complications through the cautious use of eCG and hCG in domestic and endangered nondomestic felids. PMID- 7490713 TI - Increased zona-binding ability after incubation of spermatozoa with proteins extracted from spermatozoa of fertile semen. AB - The involvement of proteins extracted from spermatozoa of fertile semen in sperm zona binding was examined under hemizona assay conditions. One droplet of a suspension of spermatozoa was exposed to sperm proteins and then tested for zona binding, while a parallel semen suspension droplet incubated with culture medium served as a control. The reliability of the test was increased by relating the number of spermatozoa bound to each inseminated hemizona to the surface area of the hemizona and expressed as the binding index. For spermatozoa incubated with extracted proteins, the binding index was greater than (P = 0.001) that of controls (125.2 +/- 45.1 versus 63.6 +/- 29.2, respectively). As a first control, two other protein sources (fetal calf serum and human follicular fluid) were tested in the hemizona assay. No significant differences were found in zona binding for other protein-exposed spermatozoa compared with controls. As a second and reverse control, exposure of one hemizona to sperm proteins before insemination with untreated spermatozoa induced a marked decrease (P = 0.0003) in sperm binding, compared with that of the matched hemizona not exposed to sperm proteins (control) (3.4 +/- 1.4 versus 74.5 +/- 6.8, respectively). Taken together, these findings confirm the involvement of extracted sperm proteins in sperm-zona interactions. Therefore, in the cases in which fertilization in vitro fails because of a lack of sperm-zona binding, incubation of deficient spermatozoa with proteins extracted from spermatozoa of fertile ejaculates should restore their ability to interact with the oocyte and, thus, should enhance the prognosis for in vitro fertilization. PMID- 7490714 TI - Differential sensitivity of ovarian follicles to gonadotrophin stimulation in broiler and layer lines of domestic fowl. AB - Differences in the reproductive potential of female broiler parent and layer fowl are related to differences in the recruitment of follicles to the preovulatory hierarchy. Broiler and laying hens were fed ad libitum or a restricted diet during rearing and ad libitum after sexual maturity. The sensitivity of thecal tissue, from ovarian follicles of different sizes, from layer and broiler fowl to stimulation by pregnant mares' serum gonadotrophin (PMSG) was determined to test the hypothesis that differences in egg production are associated with changes in the sensitivity of ovarian follicles to stimulation by gonadotrophin. The activity of ornithine decarboxylase 0.5-4 h after injection of 200 iu PMSG kg-1 body mass was assessed in thecal tissue from small white follicles 2-3 and 5-6 mm in diameter and in the smallest preovulatory yellow follicle. Basal activity was similar in layers and broiler breeders. PMSG-stimulated ornithine decarboxylase activity in follicles of 5-6 mm diameter was greater in broiler breeders than in layers. There was no difference between the lines in PMSG-stimulated ornithine decarboxylase activity in follicles of 2-3 mm diameter or in small yellow follicles. It was concluded that the low egg production and high prevalence of eggs with defective shells in broiler parents compared with layers may be related to the increased sensitivity of follicles to endogenous gonadotrophins at the critical stage of recruitment to the preovulatory hierarchy. PMID- 7490715 TI - Effect of bovine ampullary and isthmic oviductal fluid on motility, acrosome reaction and fertility of bull spermatozoa. AB - Motility, acrosome reaction and oocyte fertilizing ability were assessed for bull spermatozoa after incubation in regional (isthmic or ampullary), bovine oviductal fluid, pooled by stage of the oestrous cycle. Oviductal fluids collected daily from isthmic and ampullary cannulae implanted in the same oviduct were divided into pools, representing two oestrous cycle stages, based on daily serum progesterone concentrations. Ejaculated bull spermatozoa were incubated for 0-6 h in each type of oviductal fluid. Incubation in isthmic oviductal fluid collected during the nonluteal stage, including oestrus and ovulation, decreased overall sperm motility (from 71.7% motile spermatozoa to 34.0%) and both path (78 microns s-1 versus 86-89 microns s-1) and progressive (74 microns s-1 versus 83-85 microns s-1) velocities of spermatozoa motion. Spermatozoa incubated in isthmic, non-luteal oviductal fluid had a higher rate and extent of sperm acrosome reaction (213% of control versus 136-161% of control by 2 h incubation) compared with spermatozoa incubated in other oviductal fluid types. However, incubation in nonluteal ampullary fluid increased the number of spermatozoa, which were both acrosome reacted and live, and able to fertilize bovine ova (88.7% fertilized versus 75-81%). Glycosaminoglycan concentrations were similar among types of oviductal fluid (0.77-0.88 mg ml-1). These findings indicate that oviductal fluid differentially affects sperm function, depending on the oviduct region and the stage of the oestrous cycle at which the fluid was obtained. PMID- 7490716 TI - A novel uterine protein that associates with the embryonic capsule in equids. AB - An apparently unique protein produced in large quantities by the endometrium of the mare which adheres to, or is incorporated into, the acellular capsule that surrounds the equine conceptus in early pregnancy, has been characterized and partially sequenced. It has a molecular mass of approximately 18 kDa on SDS-PAGE gels and is nonglycosylated as assessed by a sensitive carbohydrate detection kit. Comparison of its first 24 amino-terminal amino acids with all entries in the databases failed to show any significant identity with any other protein sequence. Secretion of the protein appears to be progesterone dependent, as its presence in uterine flushings correlates with peripheral serum progesterone profiles during the oestrous cycle and its secretion can be induced in anoestrous mares by administration of a synthetic progestagen. However, in pregnant mares, the protein disappears from the uterus after about day 20 (term = 320-340 days), despite the persistence of high serum concentrations of progesterone, indicating that additional mechanisms control its synthesis and secretion. The strong association of the protein with the glycoprotein capsule that surrounds the equine blastocyst suggests that it may be incorporated into the capsule as the capsule expands from day 11 after ovulation. Alternatively, or additionally, it may be involved in the transport of nutrients or other substances through the capsule, and may therefore play an important role in the maintenance of pregnancy. PMID- 7490717 TI - Functional effects of intracellular pH alteration in the human uterus: simultaneous measurements of pH and force. AB - Changes in intracellular pH have differing effects on contraction in different types of muscle, and between species. Because of the large number of women requiring a caesarean section due to failure to progress in labour, it is important to know how the human myometrium responds to the changes in intracellular pH that may occur during labour. The pH-sensitive dye, carboxy SNARF, was used to measure intracellular pH in small strips of human myometrium. Intracellular pH and tension were simultaneously recorded in pregnant and nonpregnant tissue. Intracellular pH was altered by the addition of weak acids and bases. Intracellular alkalinization caused an increase in the frequency and amplitude of contractions. Intracellular acidification led to an initial increase in the frequency and/or the amplitude of the contractions, followed by abolition of contractions. Alterations in intracellular pH had profound effects on contraction in human uterine smooth muscle. Possible mechanisms are discussed whereby pH could influence force production, and changes in contraction are related to the speed and extent of the change in intracellular pH. PMID- 7490718 TI - Involvement of N-methyl-D-aspartate receptor using excitatory amino acid neurotransmitters in control of pulsatile secretion of LH during sexual development in Holstein bull calves. AB - The neuroendocrine mechanisms underlying the development of pulsatile release of LH in bull calves are poorly understood. The hypothesis that endogenous excitatory amino acids, working through N-methyl-D-aspartate (NMDA) receptors, are involved in the generation of pulsatile LH release during sexual maturation of bull calves was tested. Holstein bull calves were administered i.v. 0.001, 0.01 and 0.1 mg kg-1 body mass of MK-801, a specific noncompetitive NMDA receptor antagonist, on alternate days at 1, 12 and 24 weeks of age (n = 6 per dose), using a replicated 3 x 3 latin square design. Blood samples were obtained from jugular catheters at intervals of 10 min for 2 h before and 2 h after MK-801 treatment at 1 week and for periods of 4 h before and after MK-801 administration at 12 and 24 weeks of age. Plasma concentrations of LH were measured by a specific radioimmunoassay and pulsatile LH secretion was analysed using the Pulsar algorithm. Basal LH secretion at 1 week of age was low but was increased by 12 weeks (0.38 +/- 0.01 versus 1.38 +/- 0.11 ng ml-1; P < 0.001) with the establishment of frequent high amplitude pulses. Mean LH concentrations and pulse amplitude, but not frequency, were lower at 24 than at 12 weeks of age (0.88 +/- 0.08 versus 1.38 +/- 0.11 ng ml-1: P < 0.001, 1.74 +/- 0.14 versus 2.82 +/- 0.36 ng ml-1: P < 0.01, and 1.70 +/- 0.30 versus 1.70 +/- 0.41 in 4 h: P > 0.05, respectively). Administration of MK-801 did not affect LH secretion in 1- and 12 week-old calves.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7490719 TI - Plasma membrane block to polyspermy in human oocytes and preimplantation embryos. AB - The existence and time course of the human plasma membrane block to polyspermy were investigated by an in vitro fertilization assay using zona pellucida-free unfertilized oocytes, pronuclear oocytes and embryos. In the time course study using a high sperm concentration (10(5) spermatozoa ml-1), the number of penetrating spermatozoa at 30 and 60 min after insemination were 1.3 +/- 0.3 and 2.9 +/- 0.4, respectively. By 2 h after insemination, spermatozoa penetration reached a maximum. A lower maximum number of penetrating spermatozoa was observed at a low sperm concentration (10(4) spermatozoa ml-1), but the number of penetrating spermatozoa still reached a maximum by 2 h after insemination. A reinsemination experiment demonstrated that the number of penetrating spermatozoa was not significantly different between control and reinseminated oocytes, while sperm penetration was not observed in the oocyte beyond the two-cell stage. Furthermore, the number of binding spermatozoa decreased after fertilization and most of the four-cell stage embryos displayed no sperm binding. These results suggest that the plasma membrane block plays an important role in the prevention of polyspermy in the human oocyte, and that the plasma membrane block may involve permanent changes in the binding or fusion ability of spermatozoa in the oolemma after fertilization. PMID- 7490720 TI - Effects of melatonin implants in spring on testicular regression and moulting in adult male raccoon dogs (Nyctereutes procynoides). AB - Six male raccoon dogs were treated with constant-release implants of melatonin on 29 March, when the animals still had mature spermatozoa and a winter coat. Six untreated animals served as controls. High serum concentrations of melatonin were found in the treated animals throughout the study period (182.2 and 38.5 pg ml-1 in April and August, respectively). The high serum concentrations of prolactin in April and May (18.9 and 15.7 ng ml-1, respectively) in the controls were not seen in the treated animals. Testicular regression, judged by width of the testis and stage of spermatogenesis, was slowed after melatonin administration. Testis width from April to June and stage of spermatogenesis in May and June were greater in the treated animals than in the controls; however, from July onwards the differences between the two groups were no longer significant. Serum concentrations of testosterone remained low (below the detection limit in most cases) in both treated and control animals throughout the study period. Melatonin treatment also affected moulting, which was assessed by changes in the number of growing and mature underfur hairs per bundle and the percentage of bundles with a growing guard hair. The treated animals showed a more rapid shedding of mature underfur hairs and growth of new underfur hairs 6 weeks after melatonin implantation compared with the controls.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7490721 TI - Comparison of the effects of oviductal cell co-culture and oviductal cell conditioned medium on the development and metabolic activity of cattle embryos. AB - The objective of this study was to compare the development and metabolic activity of cattle embryos co-cultured with bovine oviductal cells or cultured in serum free medium previously conditioned by bovine oviductal cells. Zygotes were produced by in vitro fertilization of oocytes from bovine ovaries obtained from an abattoir. Development to the four-cell stage occurred by 48 h after fertilization in both culture systems, but co-cultured embryos reached the 16 cell stage by 96 h, whereas those cultured in conditioned medium did not do so until 24 h later. Similarly, the morula and blastocyst stages were reached 24 h earlier in co-culture than in conditioned medium. There were significantly more cells in the blastocysts from co-culture (96.8 +/- 6.1 versus 56.7 +/- 3.3; P < or = 0.0001). The metabolism of glutamine did not differ between embryos cultured in the two systems, but the metabolism of glucose was significantly greater in embryos cultured in conditioned medium. The first significant increase in glucose metabolism occurred between the four-cell and the 16-cell stages in embryos cultured in conditioned medium, but occurred between the 16-cell and morula stages in the co-cultured embryos, such that the glucose metabolism was significantly greater at the 16-cell stage in embryos cultured in conditioned medium compared with co-cultured embryos (6.5 +/- 1.0 versus 1.5 +/- 0.4 pmol per embryo per 3 h, P < or = 0.0001). The concentration of glucose was significantly less, and that of lactate significantly greater, in co-culture medium than in conditioned medium.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7490722 TI - Activation of hamster zona-free oocytes by homologous and heterologous spermatozoa. AB - Spermatozoa of a wide variety of species can fuse with zona-free hamster oocytes. Zona-free hamster oocytes were inseminated with spermatozoa of homologous (hamster) and other (mouse, guinea-pig and human) species, and their responses were closely examined to determine whether such interspecific sperm-oocyte fusion always induces normal oocyte activation. While guinea-pig and human spermatozoa could activate hamster oocytes as efficiently as hamster spermatozoa, mouse spermatozoa could not. Mouse spermatozoa fused readily with hamster oocytes, yet most oocytes remained inactivated at least during the first 1.5-2 h. The amount of M-phase (metaphase) promoting factor was reduced in hamster oocytes fused with one or several mouse spermatozoa; however, repetitive Ca2+ transients failed to occur unless oocytes were inseminated with a concentrated sperm suspension and penetrated by very many spermatozoa. These observations suggest that sperm-oocyte membrane fusion per se is not sufficient to trigger oocyte activation, and that putative sperm-derived oocyte activating factors show some degree of species specificity. PMID- 7490723 TI - New drug developments for opportunistic infections in immunosuppressed patients: Pneumocystis carinii. PMID- 7490724 TI - N-[(1-butyl-4-piperidinyl)methyl]-3,4dihydro-2H-[1,3]oxazino[3,2- a]indole10 carboxamide hydrochloride: the first potent and selective 5-HT4 receptor antagonist amide with oral activity. PMID- 7490725 TI - Synthesis and characterization of a highly potent and effective antagonist of retinoic acid receptors. PMID- 7490726 TI - (R)-(+)-N-[3-[5-[(4-fluorophenyl)methyl]-2-thienyl]-1-methyl- 2-propynyl]-N hydroxyurea (ABT-761), a second-generation 5-lipoxygenase inhibitor. AB - Structure-activity optimization of inhibitory potency and duration of action of N hydroxyurea containing 5-lipoxygenase inhibitors was conducted. The lipophilic heteroaryl template and the link group connecting the template to the N hydroxyurea pharmacophore were modified. Inhibition of 5-lipoxygenase was evaluated in vitro in a human whole blood assay. An in vitro assay using liver microsomes from monkey was used to evaluate congeners for comparative rates of glucuronidation. (3-Heteroaryl-1-methyl-2-propynyl)-N-hydroxyureas were found to be more resistant to in vitro glucuronidation. The promising inhibitor N-[3-[5-(4 fluorophenoxy)-2-furyl]-1-methyl-2-propynyl]-N- hydroxyurea (6) was found to have stereoselective glucuronidation in monkey and man. The R enantiomer 7 provided longer duration of inhibition as evaluated by an ex vivo whole blood assay. Further optimization of the lipophilic template led to the discovery of (R)-(+)-N [3-[5-[(4-fluorophenyl)methyl]-2- thienyl]-1-methyl-2-propynyl]-N-hydroxyurea (11) with more effective and prolonged inhibition of leukotriene biosynthesis than zileuton (1) and 7 in monkey and man. The optimized 5-lipoxygenase inhibitor 11 was selected for development as an investigational drug for leukotriene mediated disorders. PMID- 7490728 TI - Selective and potent monoamine oxidase type B inhibitors: 2-substituted 5 aryltetrazole derivatives. AB - Twenty new 2-(cyanoalkyl)tetrazoles (15 and 16) and twenty new 2 (hydroxyalkyl)tetrazoles (17 and 18) were synthesized and investigated in vitro for their abilities to inhibit selectively rat brain monoamine oxidase (MAO) B over MAO A. Most of them were MAO B inhibitors and those bearing a substituted 4 (arylmethoxy)phenyl group in the position 5 of the tetrazole ring had IC50 values between 8 microM for 18d and 2 nM for 16a (30 nM for lazabemide) with a selectivity toward MAO B of 37,000 for 16a. The reversibility of their inhibitory activity was demonstrated by in vitro dialysis tests. The 5-[4 (phenylmethoxy)phenyl]-2-(2-cyanoethyl)tetrazole (16a) its derivative 16h and the 5-[4-(phenylmethoxy)phenyl]-2-(2-hydroxyethyl)tetrazole (18a) and its derivative 18h were found to be potent, in vitro selective, and competitive MAO B inhibitors. Tetrazole 16a can be considered one of the most active and selective competitive MAO B inhibitors known up to now. This compound was selected for ex vivo experiments and was shown to be a strong and reversible MAO B inhibitor with a short duration of action after oral administration at 5 mg/kg. The structure activity approach gives rise to the great importance of lipophilicity over electronic effects of the compounds in these series. PMID- 7490727 TI - (E)-8-benzylidene derivatives of 2-methyl-5-(3-hydroxyphenyl)morphans: highly selective ligands for the sigma 2 receptor subtype. AB - The determination of the structure and function of the sigma receptor subtypes and their physiological role(s) has been impeded by the unavailability of selective ligands. We have developed a new class of sigma subtype selective receptor ligands that are (E)-8-benzylidene derivatives of the synthetic opioid (+/-)-, (+)-, and (-)-2-methyl-5-(3-hydroxyphenyl)morphan-7-one (1). The derivatives can be prepared by reaction of 1, (+)-1, and (-)-1 with the appropriate benzaldehyde under Claisen-Schmidt conditions. Incorporation of substituted (E)-8-benzylidene moieties onto the 7-keto precursor of (+)-2-methyl 5-(3-hydroxyphenyl)morphan, (+)-1, produces compounds (-)-2 through (-)-7 (5.8 32.0 nM, sigma 1), which have between a 25- and 131-fold increase in affinity for the sigma 1 receptor subtype relative to the keto precursor (+)-1 (Ki = 762 nM, sigma 1). Compound (-)-2 is the most selective of this group (16-fold) for the sigma 1 subtype versus sigma 2. Substitution of an (E)-8-benzylidene moiety onto the 7-keto precursor of (-)-2-methyl-5-(3-hydroxyphenyl)morphan, (-)-1, produces compounds (+)-2-(+)-9 (6.4-52.6 nM, sigma 2), which have at least a 475-3906-fold increase in affinity for the sigma 2 receptor subtype relative to the keto precursor (-)-1 (Ki = 25 x 10(3) nM). This enhancement of sigma 2 receptor affinity is accompanied by substantial selectivity of all of these dextrorotatory products for the sigma 2 relative to the sigma 1 subtype (32-238-fold), and thus, they are among the most sigma 2 selective compounds currently known. Furthermore, the sigma 1 subtype is highly enantioselective for the levorotatory isomers, (-) 2-(-)-7 (41-1034-fold), whereas the sigma 2 subtype is only somewhat enantioselective for the dextrorotatory isomers, (+)-2-(+)-7 (2.6-9.3-fold). All of these derivatives retain substantial affinity for the mu opioid receptor. Despite the high affinity of the dextrorotatory derivatives for the mu opioid receptor, the high affinity and selectivity for sigma 2 over sigma 1 sites will surely prove beneficial as tools for the delineation of the function and physiological role of sigma 2 receptors. PMID- 7490729 TI - Identification of a series of 3-(benzyloxy)-1-azabicyclo[2.2.2]octane human NK1 antagonists. AB - The synthesis and in vitro and in vivo evaluation of a series of 3-(benzyloxy)-1 azabicyclo-[2.2.2]octane NK1 antagonists are described. While a number of 3,5 disubstituted benzyl ethers afford high affinity, the 3,5 bis(trifluoromethyl)benzyl was found to combine high in vitro affinity with good oral activity. Detailed structure-activity relationship studies in conjunction with data from molecular modeling and mutagenesis work have allowed the construction of a model of the pharmacophore. Specific interactions that have been identified include an interaction between His-197 and one of the rings of the benzhydryl, a lipophilic pocket containing His-265 that the benzyl ether occupies, and a possible hydrogen bond between Gln-165 and the oxygen of the benzyl ether. PMID- 7490730 TI - N-3-substituted pyrimidinones as potent, orally active, AT1 selective angiotensin II receptor antagonists. AB - A novel series of nonpeptide angiotensin II (A II) antagonists containing a pyrimidinone ring which carries a C-linked biphenyltetrazole moiety and a carboxyheteroaryl group on the 3-position have been prepared. Their affinity for the AT1 receptor was determined in a binding assay on rat adrenal cortical membranes. The in vivo antihypertensive properties were tested by evaluating the inhibition of the pressor response to A II followed by iv and id administration. Extensive molecular modeling studies, including comparison of molecular electrostatic potential distributions, conformational analysis, and overlays on a computational pharmacophore model of A II, were used to evaluate structural parameters of the new compounds, in comparison to other known A II antagonists (e.g., DUP-753 and SK&F 108566). According to the modeling studies, the introduction of a (carboxyheteroaryl)methyl moiety at the 3-position of the pyrimidinone ring led to derivatives with increased potency. Methyl 2-[[4-butyl-2 methyl-6-oxo-5-[[2'-(1H-tetrazol-5-yl)[1,1'-biphenyl ]- 4-yl]methyl]-1-(6H) pyrimidinyl]methyl]-3-thiophenecarboxylate (3k, LR-B/081), one of the most potent compounds in the series (Ki = 1.4 nM), exhibited a marked antihypertensive activity on oral administration to conscious renal hypertensive rats, with long duration of action. It was selected for clinical evaluation in the treatment of hypertension in man. PMID- 7490731 TI - Synthesis and structure-activity relationships of acetylcholinesterase inhibitors: 1-benzyl-4-[(5,6-dimethoxy-1-oxoindan-2-yl)methyl]piperidine hydrochloride and related compounds. AB - Following the discovery of a new series of anti-acetylcholinesterase (anti-AChE) inhibitors such as 1-benzyl-4-[2-(N-benzoylamino)ethyl]piperidine (1), we reported that its rigid analogue, 1-benzyl-4-(2-isoindolin-2-ylethyl)piperidine (5), had more potent activity. We have extended the structure-activity relationship (SAR) study for the rigid analogue and found that the 2-isoindoline moiety in compound 5 can be replaced with a indanone moiety (8) without a major loss in potency. Among the indanone derivatives, 1-benzyl-4-[(5,6-dimethoxy-1 oxoindan-2-yl)methyl]piperidine (13e) (E2020) (IC50 = 5.7 nM) was found to be one of the most potent anti-AChE inhibitors. Compound 13e showed a selective affinity 1250 times greater for AChE than for butyrylcholinesterase. In vivo studies demonstrated that 13e has a longer duration of action than physostigmine at a dose of 5 mg/kg (po) and produced a marked and significant increase in acetylcholine content in rat cerebral cortex. We report the synthesis, SAR, and a proposed hypothetical binding site of 13e (E2020). PMID- 7490732 TI - Novel non-nucleoside inhibitors of human immunodeficiency virus type 1 (HIV-1) reverse transcriptase. 4. 2-Substituted dipyridodiazepinones as potent inhibitors of both wild-type and cysteine-181 HIV-1 reverse transcriptase enzymes. AB - The major cause of viral resistance to the potent human immunodeficiency virus type 1 reverse transcriptase (RT) inhibitor nevirapine is the mutation substituting cysteine for tyrosine-181 in RT (Y181C RT). An evaluation, against Y181C RT, of previously described analogs of nevirapine revealed that the 2 chlorodipyridodiazepinone 16 is an effective inhibitor of this mutant enzyme. The detailed examination of the structure-activity relationship of 2-substituted dipyridodiazepinones presented below shows that combined activity against the wild-type and Y181C enzymes is achieved with aryl substituents at the 2-position of the tricyclic ring system. In addition, the substitution pattern at C-4, N-5, and N-11 of the dipyridodiazepinone ring system optimum for inhibition of both wild-type and Y181C RT is no longer the 4-methyl-11-cyclopropyl substitution preferred against the wild-type enzyme but rather the 5-methyl-11-ethyl (or 11 cyclopropyl) pattern. The more potent 2-substituted dipyridodiazepinones were evaluated against mutant RT enzymes (L100I RT, K103N RT, P236L RT, and E138K RT) that confer resistance to other non-nucleoside RT inhibitors, and compounds 42, 62, and 67, with pyrrolyl, aminophenyl, and aminopyridyl substituents, respectively, at the 2-position, were found to be effective inhibitors of these mutant enzymes also. PMID- 7490733 TI - Novel non-nucleoside inhibitors of human immunodeficiency virus type 1 reverse transcriptase. 5. 4-Substituted and 2,4-disubstituted analogs of nevirapine. AB - Molecular modeling analysis of the recently published X-ray crystal structure of nevirapine bound to wild type human immunodeficiency virus type 1 reverse transcriptase (WT-RT) indicated the presence of a lipophilic cavity proximal to the 4-position of the inhibitor. A series of 4-substituted derivatives of nevirapine were thus synthesized to assess structure-activity relationships (SARs) and to see if increased binding to this region might translate into greater activity against mutant RTs. The results show that compounds with an appropriately spaced aryl ring appended to the 4-position of the dipyridodiazepinone ring system show good activity against WT-RT. Furthermore certain derivatives appear to inhibit the Y181C mutant RT. Attempts to combine these results with the recent discovery that 2-substituents enhance activity against the Y181C mutant led to a few compounds with moderate activity against both enzymes. The SAR of these two positions, however, could not be combined in a simple fashion. PMID- 7490734 TI - Design and synthesis of conformationally constrained analogues of 4-(3-butoxy-4 methoxybenzyl)imidazolidin-2-one (Ro 20-1724) as potent inhibitors of cAMP specific phosphodiesterase. AB - The synthesis and biological evaluation of cAMP-specific phosphodiesterase (PDE IV) inhibitors is described. The PDE IV inhibitor 4-(3-butoxy-4 methoxybenzyl)imidazolidin-2-one (Ro 20-1724, 2) was used as a template from which to design a set of rigid oxazolidinones, imidazolidinones, and pyrrolizidinones that mimic Ro 20-1724 but differ in the orientation of the carbonyl group. The endo isomer of each of these heterocycles was more potent than the exo isomer in an enzyme inhibition assay and a cellular assay, which measured TNF alpha secretion from activated human peripheral blood monocytes (HPBM). Imidazolidinone 4a inhibited human PDE IV with a Ki of 27 nM and TNF alpha secretion from HPBM with an IC50 of 290 nM. By comparison, Ro 20-1724 is significantly less active in these assays with activities of 1930 and 1800nM, respectively. PMID- 7490735 TI - Myosin from alga Chara: unique structure revealed by electron microscopy. AB - Cytoplasmic streaming is a fundamental process for the transport of molecules and organelles in plant cells. A myosin-like protein responsible for cytoplasmic streaming in fresh water alga, Chara corallina, has been purified. Electron micrographs of this protein revealed that it has two heads with similar size and shape to those of muscle myosin (myosin II) but its tail was shorter than that of muscle myosin with a globular structure at its distal end like that of myosin V. The myosin cosedimented with vesicles composed of phosphatidyl serine like other non-muscle myosin species involved in intracellular transport. PMID- 7490736 TI - Examination of the structure of the transthyretin amyloid fibril by image reconstruction from electron micrographs. AB - Familial amyloidotic polyneuropathies are autosomal-dominant, inherited disorders that are characterised by the aggregation of variant proteins in a fibrillar form and by the extracellular deposition of amyloid fibrils. In familial amyloidotic polyneuropathy type I the protein constituent is a variant transthyretin molecule that has a Val to Met substitution at residue 30. Patients with this form of the disease present with sensory and motor disturbances, widespread autonomic dysfunction and in some cases, vitreous opacities. We have used amyloid material from the vitreous humours of patients homozygous for this mutation and analysed the structure of the fibrils by thin section electron microscopy and image reconstruction. Cross-sectional images of 200 different fibrils were collected and aligned, manually at first and then with an automated process that uses iterative cross-correlation. The averaged cross-section calculated produced a detailed view of the fibril substructure. The diameter of the fibrils is about 130 A. In cross-section they exhibit 4-fold symmetry with four proto-filaments, each measuring 40 to 50 A across, arranged around a central hollow core. PMID- 7490737 TI - Disparate replication properties of integrated and extrachromosomal forms of bovine papilloma virus in ID13 cells. AB - Bovine papillomavirus (BPV) previously has been reported to exist in transformed rodent cell lines as both chromosomally integrated and extrachromosomal forms. In the BPV-transformed mouse cell line ID13, extrachromosomal BPV molecules replicate throughout S phase of the cell cycle in a random choice mode. We report here that these replication properties were altered for chromosomally integrated BPV DNA in five independent ID13 subclones. In all of the subclones, the integrated BPV sequences, which had no detectable deletions or mutations, existed in head-to-tail tandem arrays that replicated once per cell cycle, predominantly late in S phase. In contrast, extrachromosomal BPV molecules present in other subclones of the same cell line replicated in the random choice mode observed previously for non-integrated BPV. Our results indicate that the replication origin of integrated BPV either is inactivated as a consequence of chromosomal insertion, leading to the replication of BPV from origins in the flanking chromosomal DNA, or alternatively is reprogrammed to function in a once-per-cell cycle mode predominantly late in S phase. PMID- 7490738 TI - Mechanism of protein access to specific DNA sequences in chromatin: a dynamic equilibrium model for gene regulation. AB - We present evidence for a mechanism by which regulatory proteins may gain access to their target DNA sequences in chromatin. In this model, nucleosomes are dynamic structures, transiently exposing stretches of their DNA. Regulatory proteins gain access to DNA target sites in the exposed state, and bind with an apparent dissociation constant equal to their dissociation constant for naked DNA divided by a position-dependent equilibrium constant for site exposure within the nucleosome. A sensitive assay, based on the kinetics of restriction digestion of sites within nucleosomes, reveals this dynamic behaviour and quantifies the equilibrium constants for site exposure. Our results have implications for many aspects of chromatin function. They offer new mechanisms for cooperativity (synergy) in regulatory protein binding and for active invasion of nucleosomes. PMID- 7490739 TI - The DNA-binding domain of the hexameric arginine repressor. AB - The arginine repressor of Escherichia coli is a classical feedback regulator, signalling the availability of L-arginine inside the cell. It differs from most other bacterial repressors in functioning as a hexamer, but structural details have been lacking and its shares no clear sequence homologies with other transcriptional regulators. Analysis of the amino acid residue sequence and proteolytic cleavage pattern of the repressor was used to identify a region predicted to house the DNA-binding function. When this protein fragment is overexpressed from a clone of the corresponding gene fragment, it represses ornithine transcarbamylase levels in vivo, and binds to the operator DNA in vitro, both in an arginine-independent manner. Sedimentation equilibrium and gel filtration indicate that the purified protein fragment is a monomer in solution. The results thus define the domain organization of the repressor at low resolution, suggesting that the N and C-terminal portions of the polypeptide chain are separated by a structural and functional border that decouples hexamerization and arginine binding from DNA binding. PMID- 7490740 TI - An interactive framework for RNA secondary structure prediction with a dynamical treatment of constraints. AB - A novel approach aiding in the prediction of RNA secondary structures is presented. Although phylogenetic methods are the most successful at deriving RNA secondary structures, the are not applicable when the number of sequences or the sequence variability is too low. Methods based on energy minimization are therefore of great interest. However, some of the suboptimal RNA secondary structures computed with classic methods are unsaturated structures, i.e. some structures are included into others. Thus, the incorporation of constraints during the process of folding is not possible, while the incorporation of constraints before the process of folding often introduces a bias into the energy function. This paper describes a new procedure which allows for the incorporation of constraints before and during the process of RNA folding. SAPSSARN is an interactive program which offers a framework, both to specify a secondary structure through a set of folding constraints and to compute all the supoptimal saturated RNA secondary structures which satisfy all the folding constraints. At the start, it relies on the computation of the probabilities of pairing of each base with all others according to McCaskill's algorithm. The constraint satisfaction formulation of the problem deals dynamically with a chosen set of folding constraints and, finally, a search algorithm computes all the suboptimal saturated secondary structures which satisfy those folding constraints. Within such a framework, it is possible to test new ideas about RNA folding and secondary structures, including pseudoknots, can be computed. The program is illustrated with RNA sequences on which we obtained results in agreement with known structures by using a protocol which mimics the hierarchical folding of RNA molecules. PMID- 7490741 TI - Mapping of the active site zinc ligands of peptide deformylase. AB - A set of 50 site-directed mutants of the Escherichia coli fms gene was constructed to delineate the residues of the active site of peptide deformylase, including the ligands of the zinc ion. In particular, because zinc is usually coordinate by Asp, Cys, Glu or His residues, all the corresponding codons were individually changed. The functional consequence of the substitutions was assessed by complementation of a fms-null strain with the help of vectors expressing the mutate genes. In addition to the mutations of the Cys90 codon, only those of the three conserved residues of the 132HEXXH136 motif of peptide deformylase prevented the indicator strain growing. Most enzyme variants were purified to homogeneity in a second step. Their characterization in vitro showed that the defects in complementation as observed in vivo corresponded to huge decreases of deformylation efficiency. The change of Glu88 also led to a significant decrease in catalytic rate. Unexpectedly, upon substitutions of Glu79 or of Glu83, the enzymes exhibited a strongly increased catalytic efficiency. The measurement of the content of zinc in each purified variant indicated that Cys90, His132 and His136 bound the metal ion. Zinc-free variants mutated at these positions were obtained and shown to display an increased sensitivity to proteolytic attack. Altogether, the data showed that both the presence of zinc and the conserved residues of the HEXXH motif were crucial for the activity of deformylase. This behaviour identified the enzyme as a member of the zinc metalloproteases superfamily. However, the unexpected participation in the binding of the zinc atom of Cys90, upstream from the HEXXH motif, suggested that peptide deformylase could be representative of a new sub-family, distinct from those of thermolysin and astacin. PMID- 7490743 TI - Crystal structure of diferric hen ovotransferrin at 2.4 A resolution. AB - The three-dimensional structure of diferric hen ovotransferrin has been determined by X-ray crystallography at 2.4 A resolution. The structure was solved by molecular replacement, using the coordinates of diferric human lactoferrin as a search model. Several rounds of simulated annealing and restrained least squares refinement have resulted in a model structure with an R-factor of 0.171 for the data between 11.0 and 2.4 A resolution. The model comprises 5284 protein atoms (residues 5 to 686), 2 Fe3+, 2 CO3(2)- and 132 water molecules. The overall structure of ovotransferrin is similar to those of human lactoferrin and rabbit serum transferrin, being folded into two homologous lobes, each containing two dissimilar domains with one Fe3+ and one CO3(2)- bound at a specific site in each interdomain cleft. However, the relative orientation of the two lobes, which may be related to the class specificity of transferrins to receptors, is different from either human lactoferrin or rabbit serum transferrin. The angle of the relative orientation in ovotransferrin is increased by 6.8 degrees and 15.7 degrees as compared with to those in rabbit serum transferrin and human lactoferrin, respectively. Interdomain Lys209-Lys301 and Gln541-Lys638 interactions are found near the metal binding site of each lobe. The interlobe interactions and their role in the stabilization of iron binding are discussed. PMID- 7490744 TI - Three-dimensional structures of the Fab fragment of murine N1G9 antibody from the primary immune response and of its complex with (4-hydroxy-3-nitrophenyl)acetate. AB - The three-dimensional structures of the Fab fragment, in its unliganded and liganded crystals, of mouse anti-(4-hydroxy-3-nitrophenyl)acetate (NP) antibody N1G9 have been determined by the molecular replacement method. The unliganded and NP-liganded structures were refined at 2.4 A resolution to crystallographic R factors of 0.194 and 0.196, respectively. Antibody N1G9 bears lambda light chains, and is one of the primary immune response antibodies. Fab N1G9 exhibits an elbow angle of 197 degrees in both structures. This large angle is ascribed to the VL-CL interface formed by lambda-chain residues. A hydrophobic pocket surrounded by the complementarity-determining regions except L2 is identified as a hapten-binding site. Between the liganded and unliganded structures, root-mean square (r.m.s.) positional deviations are 0.42 A for the main-chain atoms, and 0.74 A for all the protein atoms. The major structural differences between these structures are localized in the hapten-binding site, and yield an r.m.s. deviation of 1.03 A for the side-chain atoms. The soaked NP ligand is in van der Waals contact with the aromatic side-chains of Tyr32L and Trp91L of the light chain, and Trp33H and Tyr97H of the heavy chain, and is hydrogen-bonded to the side-chains of Trp96L, His35H, Arg50H, Tyr95H, and Ser100aH. The side-chain of Lys58H is salt-bridged to the NP hydroxyl group. The side-chains of Arg50H, Trp33H, and Tyr97H are shifted toward the NP carboxyl group. The side-chain of Trp33H, whose replacement to Leu increases affinity by tenfold, is sandwiched between the Arg50H and Tyr97H side-chains, and is in cramped contact both with the ligand and with these side-chains. Affinity increases in the maturation of the anti-NP antibodies are ascribable to conformational relief of these cramped contacts through the replacement of Trp33H or through suitable structural alterations in the H3 region. PMID- 7490742 TI - Genomic organization and chromosomal location of the human dsRNA adenosine deaminase gene: the enzyme for glutamate-activated ion channel RNA editing. AB - The structure of the human gene encoding the double-stranded RNA (dsRNA) adenosine deaminase (DRADA) was characterized. This nuclear localized enzyme is involved in the RNA editing required for the expression of certain subtypes of glutamate-gated ion channel subunits. The DRADA gene span 30 kb pairs and harbors 15 exons. The transcription of the DRADA gene driven by the putative promoter region, which contains no typical TATA or CCAAT box-like sequences, is initiated at multiple sites, 164 to 216 nucleotides upstream of the translation initiation codon. The three dsRNA binding motifs (DRBM), 70 amino acid residues long, are each encoded by two exons plus an intervening sequence that interrupts the motif at the identical amino acid position. This finding is consistent with the notion that the dsRNA binding domains may be composed of two separate functional subdomains. Fluorescent in situ hybridization localized the DRADA gene on the long arm chromosome 1, region q21. The gene structure and sequence information reported in this study will facilitate the investigation of involvement of DRADA in hereditary diseases that may be the result of malfunction of glutamate-gated ion channels. PMID- 7490745 TI - The refined crystallographic structure of a DD-peptidase penicillin-target enzyme at 1.6 A resolution. AB - The D-alanyl-D-alanine peptidase from Streptomyces sp. R61 is a 37,500 dalton exocellular enzyme that has served as a model for membrane-bound peptidases that are involved in bacterial cell wall biosynthesis. Inhibition of these enzymes by beta-lactam antibiotics ultimately leads to bacterial cell death. The X-ray crystal structure of the R61 D-alanyl-D-alanine peptidase has been solved using multiple isomorphous replacement, simulated annealing and least squares refinement. The space group and unit cell parameters are P2(1)2(1)2(1) with a = 51.1 A, b = 67.3 A and c = 102.4 A. The structure has been refined using 2 sigma data to 1.6 A resolution with a crystallographic R-factor of 0.148. The model contains 347 residues (2938 atoms) and 254 solvent molecules. The overall temperature factor is 9.6 A2, and the estimated coordinate error is 0.14 A. The protein consists of a single polypeptide chain organized into two regions. One region contains a nine-stranded antiparallel beta-sheet with helices on both faces; this region includes both the amino and carboxyl termini. The second region is all helical. Sixty percent of the residues occur in helices or beta sheet. The reactive Ser62 is found between the two regions of the enzyme at the amino end of the protein's longest-helix which begins with one turn of 3(10) helix and continues with four turns of alpha-helix. The active site is an elongated pocket that contains four basic and four aromatic residues. An oxyanion hole is formed by Ser62 NH and Thr301 NH. The pocket also contains the few key residues that are conserved in all penicillin-binding proteins and beta lactamases. Two of these residues, Lys65 and Tyr159, are among the 16 side-chains that take on multiple conformations in the R61 crystal structure. Three of the 12 proline rings adopt two conformations which we believe has not been previously reported. There is no anionic acid equivalent to the catalytic Glu166 found in Class A beta-lactamases. Two ordered water molecules (O507 and O644) are found buried in the active site and hydrogen-bonded to each other (2.6 A). O507 could potentially act as the hydrolytic water molecule for deacylation. PMID- 7490746 TI - Crystal structure of concanavalin B at 1.65 A resolution. An "inactivated" chitinase from seeds of Canavalia ensiformis. AB - Seeds of Canavalia ensiformis (jack bean) contain besides large amounts of canavalin and concanavalin A, a protein with a molecular mass of 33,800 which has been named concanavalin B. Although concanavalin B shares about 40% sequence identity with plant chitinases belonging to glycosyl hydrolase family 18, no chitinase activity could be detected for this protein. To resolve this incongruity concanavalin B was crystallised and its three-dimensional structure determined at 1.65 A (1 A = 0.1 nm) resolution. The structure consists of a single domain with a (beta/alpha)8 topology. A 30 amino acid residue long loop occurs between the second beta-strand of the barrel and the second alpha-helix. This extended loop is unusual for the (beta/alpha)8 topology, but appears in a similar conformation in the structures of the seed protein narbonin and several chitinases as well. Two non-proline cis-peptide bonds are present in the structure of concanavalin B: Ser34-Phe, and Trp265-Asn. This structural feature is rarely observed in proteins, but could also be identified in the three dimensional structures of family 18 chitinases and narbonin in coincident positions. In the chitinases the aromatic residues of the non-proline cis peptides have been proposed to have a function in the binding of the substrate. The region in concanavalin B, where in chitinases the active site is located, shows two significant differences. First, the catalytic glutamic acid is a glutamine in concanavalin B. Second, although part of the substrate binding cleft of the chitinases is present in concanavalin B, it is much shorter. From this we conclude that concanavalin B and family 18 chitinases are closely related, but that concanavalin B has lost its enzymatic function. It still may act as a carbohydrate binding protein, however. PMID- 7490747 TI - Solution structure of the ribosome-binding domain of E. coli translation initiation factor IF3. Homology with the U1A protein of the eukaryotic spliceosome. AB - Initiation of translation in prokaryotes requires the formation of a complex between the messenger RNA, the 30 S ribosomal subunit and the initiator tRNA(fMet). Initiation factor IF3 binds to the 30 S ribosomal subunit and proof reads the initiation complex, thereby ensuring the accuracy of this step. IF3 also plays a pleiotropic role in the regulation of translation, as a result of differential influences exerted on the levels of the initiation of translation of genes or groups of genes. IF3 is composed of two independent domain or roughly identical sizes. We have expressed and purified the C-terminal domain of E. coli IF3 and shown that it retains both the 30 S particle binding and 70 S ribosome dissociating activities of the native protein. We have obtained 1H and 15N NMR resonance assignments and its 3D solution structure was calculated using 551 restraints. It is composed of a mixed beta-sheet backed by two alpha-helices. It shows a striking resemblance to the U1A small nuclear ribonucleoprotein structure, which binds to the U1 snRNA in the eukaryotic spliceosome. This suggests a convergent evolution process for these two proteins that are associated with ribonucleoproteic complexes. PMID- 7490748 TI - The structure of the transition state for folding of chymotrypsin inhibitor 2 analysed by protein engineering methods: evidence for a nucleation-condensation mechanism for protein folding. AB - The 64-residue protein chymotrypsin inhibitor 2 (CI2) is a single module of structure. It folds and unfolds as a single co-operative unit by simple two-state kinetics via a single rate determining transition state. This transition state has been characterized at the level of individual residues by analysis of the rates and equilibria of folding of some 100 mutants strategically distributed at 45 sites throughout the protein. Only one residue, a helical residue (Ala16) buried in the hydrophobic core, has its full native interaction energy in the transition state. The only region of structure which is well developed in the transition state is the alpha-helix (residues 12 to 24). But, the interactions within it are weakened, especially at the C-terminal region. The rest of the protein has varying degrees of weakly formed structure. Thus, secondary and tertiary interactions appear to form concurrently. These data, reinforced by studies on the structures of peptide fragments, fit a "nucleation-condensation" model in which the overall structure condenses around an element of structure, the nucleus, that itself consolidates during the condensation. The high energy transition state is composed of the whole of the molecule making a variety of weak interactions, the nucleus being those residues that make the strongest interactions. The nucleus here is part of the alpha-helix and some distant residues in the sequence with which it makes contacts. The remainder of the protein has to be sufficiently ordered that it provides the necessary interactions to stabilize the nucleus. The nucleus is only weakly formed in the denatured state but develops in the transition state. The onrush of stability as the nucleus consolidates its local and long range interactions is so rapid that it is not yet fully formed in the transition state. The formation of the nucleus is thus coupled with the condensation. These results are consistent with a recent simulation of the folding of a computer model protein on a lattice which is found to proceed by a nucleation-growth mechanism. We suggest that the mechanism of folding of CI2 may be a common theme in protein folding whereby fundamental folding units of larger proteins, which are modelled by the folding of CI2, form by nucleation-condensation events and coalesce, perhaps in a hierarchical manner. PMID- 7490749 TI - Search for nucleation sites in smaller fragments of chymotrypsin inhibitor 2. AB - There is a region of well-ordered structure in the transition state of folding of chymotrypsin inhibitor 2 (CI2) that consists of N-terminal residues in the unique alpha-helix (residues 12 to 24) plus some long range interactions, in particular those of Ala16 with Ile57 and Leu49 in the hydrophobic core. This is proposed to be a nucleation site. A crucial question for understanding the initiation of protein folding is: when is the nucleation site formed? Is the alpha-helix pre formed in the nominally unfolded state, or does it require long-range interactions to be stabilized? To answer this question, we have characterized a series of N-terminal fragments of CI2, each containing an increasing number of subsets of the regular secondary structure. Four small fragments have been examined by circular dichroism and two-dimensional 1H and 15N NMR spectroscopy. The smallest, [1-5], comprises the sequence corresponding to the first beta strand of the intact protein; the second, [1-13], contains also a type III reverse turn, the second beta-strand, and a type II reverse turn; the third [1 25], consists additionally of the sequence corresponding to the alpha-helix (residues 12 to 24); the fourth, [1-28], contains, in addition, the turn following the alpha-helix. All the fragments have disordered, non-compact structure in aqueous solution. In the structure-promoting co-solvent, trifluoroethanol, alpha-helical structure is stabilized in [1-25] and [1-28] in the region corresponding to the alpha-helix in the intact protein; however, the helix is frayed at both ends and is only fractionally populated, being in dynamic equilibrium with extended conformations. These observations indicate that there is little drive for independent formation of local secondary structure in CI2, and this is reflected in the highly concerted nature of the folding reaction of this protein. The nucleation site of folding of CI2 does not accumulate in the starting state for the folding reaction, but remains embryonic until there are sufficient long range interactions to stabilize it. PMID- 7490750 TI - A comparison of the pH, urea, and temperature-denatured states of barnase by heteronuclear NMR: implications for the initiation of protein folding. AB - The denatured states of barnase that are induced by urea, acid, and high temperature and acid have been assigned and characterised by high resolution heteronuclear NMR. The assignment was completed using a combination of triple resonance and magnetisation-transfer methods. The latter was facilitated by selecting a suitable mutant of barnase (Ile-->Val51) which has an appropriate rate of interconversion between native and denatured states in urea. 3J NH-C alpha H coupling constants were determined for pH and urea-denatured barnase and intrinsic "random coil" coupling constants are shown to be different for different residue types. All the denatured states are highly unfolded. But, a consistent series of weak correlations in chemical shift, NOESY and coupling constant data provides evidence that the acid-denatured state has some residual structure in regions that form the first and second helices and the central strands of beta-sheet in the native protein. The acid/temperature-denatured states has less structure in these regions, and the urea-denatured state, less still. These observations may be combined with detailed analyses of the folding pathway of barnase from kinetic studies to illuminate the relevance of residual structure in the denatured states of proteins to the mechanism of protein folding. First, the folding of barnase is known to proceed in its later stages through structures in which the first helix and centre of the beta-sheet are extensively formed. Thus, embryonic initiation sites for these do exist in the denatured states and so could well develop into true nuclei. Second, it has been clearly established that the second helix is unfolded in these later states, and so residual structure in this region of the protein is non-productive. These data fit a model of protein folding in which local nucleation sites are latent in the denatured state and develop only when they make interactions elsewhere in the protein that stabilise them during the folding process. Thus, studies of the structure of denatured states pinpoint where nucleation sites may be, and the kinetic and protein engineering studies show which ones are productive. PMID- 7490751 TI - Comparison between the phi distribution of the amino acids in the protein database and NMR data indicates that amino acids have various phi propensities in the random coil conformation. AB - It has been indicated that amino acids have various intrinsic phi and psi propensities, as demonstrated from the comparison between experimental secondary structure propensities and their relative statistical distribution in the protein database for the appropriate region of the Ramachandran plot. However, this does not eliminate the possibility that these experimental propensities are the result of context effects due to the secondary structure environment of the mutated position. To demonstrate that there are at least real intrinsic phi propensities, independent of context effects, we have used two different nuclear magnetic resonance parameters related to the phi dihedral angle (J3 alpha HN coupling constants and the chemical shift of the C alpha H proton), determined in random coil tetra- and pentapeptides, and/or in proteins. Comparison of the experimentally determined values for these parameters with the theoretical ones determined from the analysis by different empirical and theoretical equations of the phi dihedral angle statistical distribution of the amino acids in the protein database, supports the idea that each amino acid has, at least, different phi intrinsic propensities. Consideration of all conformations, or only coil conformations, in the protein database produces similar results. The reasonable correlation between these experimental and theoretical data and the hydrogen exchange data in random-coil peptides suggests that maximisation of hydrophobic surface-buried and hydrogen-bond formation with the solvent could be responsible for these different random-coil conformational preferences. Analysis of the intrinsic propensities for beta-strand, alpha-helix and polyproline II dihedral angles of the 20 amino acids in coil conformations, indicates that the side-chain of the amino acids is mainly determining the relative preferences for the phi angle. PMID- 7490752 TI - The gene for nucleoside diphosphate kinase functions as a mutator gene in Escherichia coli. AB - Nucleoside diphosphate (NDP) kinase is a key enzyme in the control of cellular concentrations of nucleoside triphosphates, and has been shown to play important roles in various cellular activities such as developmental control, signal transduction and metastasis in eukaryotic systems. In this study, the gene for NDP kinase of Escherichia coli (ndk) was disrupted and surprisingly found to be dispensable without any discernible effects on cell growth or morphology. However, a mutator phenotype was found in ndk-disruption strains; frequencies of spontaneous mutations to rifampicin resistance and nalidixic acid resistant significantly increased. A higher frequency in reversion mutations was observed with use of an amber mutation in the kanamycin-resistance gene in an ndk disruption strain. Imbalance in dNTP pools, in particular a significant increase of the dCTP content was observed, which is likely to result in the higher spontaneous mutation rates. These results suggest that NDP kinase, although not essential, plays an important role in the appropriate balance of intracellular dNTP pools to maintain a high DNA replication fidelity. Strains with ndk- pykA- pykF- as well as ndk- scs- were constructed without any discernible effect on cell growth, indicating that there is yet another enzyme(s) catalyzing nucleoside triphosphate synthesis, in addition to NDP kinase, pyruvate kinases and succinyl CoA synthetase. PMID- 7490753 TI - Functional map of the alpha subunit of Escherichia coli RNA polymerase: amino acid substitution within the amino-terminal assembly domain. AB - The alpha subunit of Escherichia coli RNA polymerase plays a key role in assembly of the core enzyme. In previous studies the amino-terminal domain consisting of 215 amino acid residues between positions 21 and 235 was identified to be involved in this assembly, and the sites for beta and beta' association were suggested to be located within or near the two conserved regions in this amino terminal assembly domain of alpha. For detailed functional mapping, Ala was substituted for 26 highly conserved amino acids around residues 40, 80 and 170 to 210. The alpha-point mutants were analyzed in vitro for their abilities to form dimers and to assemble beta beta' subunits. New types of assembly-deficient mutants were identified: alpha-R45A (having substituted Ala for Arg at residue 45) dimerized but did not assemble beta (and beta') subunits; and alpha-L48A showed a decreased level of alpha 2 beta subassembly formation, indicating that this region (residues 45 to 48) is responsible for beta-binding. Isolation of two mutants, alpha-K86A and alpha-V173A, both forming alpha 2 beta but not alpha 2 beta beta' complex, confirmed our previous conclusion that two separated regions participate in beta'-binding. PMID- 7490754 TI - Tat functions to stimulate the elongation properties of transcription complexes paused by the duplicated TAR RNA element of human immunodeficiency virus 2. AB - In this study we have defined the in vitro requirements for transcriptional regulation of the HIV-2 LTR in response to the HIV-1 and HIV-2 Tat proteins and addressed potential mechanisms of Tat function. HIV-2 contains a duplicated TAR RNA stem-loop structure in contrast to the single stem-loop structure found in HIV-1 TAR RNA. We demonstrated that the HIV-2 proximal TAR RNA stem-loop structure was more important for in vitro transcriptional activation by the HIV-1 and HIV-2 Tat proteins than the distal TAR RNA stem-loop though this downstream TAR element itself was able to confer Tat-responsiveness. The role of the two HIV 2 TAR RNA stem-loop bulge sequences was less critical than the loop sequences for in vitro transcriptional activation by Tat. In addition, we demonstrated that replacing the HIV-2 TATA element with that of HIV-1 markedly reduced the overall level of Tat activation. The role of the Tat-1 and Tat-2 proteins on the synthesis of HIV-1 and HIV-2 promoter proximal and promoter distal transcripts was then investigated. In contrast to the HIV-1 promoter, the HIV-2 promoter generated abundant levels of short transcripts in vitro transcription assays likely due to the structure of its duplicated TAR element. Both Tat-1 and Tat-2 increased the level of transcripts extending to the end of the HIV-1 and HIV-2 TAR elements as well as the level of transcripts extending more than 500 nucleotides from the transcription initiation site. However, the synthesis of transcripts within 30 nucleotides of the HIV-2 LTR transcription initiation site was unchanged in either the presence or absence of Tat while the level of transcripts extending increasing distances from the HIV-2 LTR transcription initiation site were progressively stimulated in the presence of Tat. Though the HIV-1 Tat protein was a stronger inducer of HIV-1 LTR transcription than the HIV 2 Tat protein, we did not detect differences in the binding of these proteins to the HIV-1 and HIV-2 TAR RNAs. This suggested that differences in their transactivation properties may be due to alterations in their association with RNA polymerase II or associated elongation factors. (ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7490755 TI - Electron microscopic visualization of RecT protein and its complexes with DNA. AB - Electron microscopy has been used to examine Escherichia coli RecT protein alone and in the complexes it forms with DNA substrates, with which it catalyzes strand exchange in vitro. Negative staining has revealed that the 33 kDa RecT protein monomers form open C-shaped and closed O-shaped particles. RecT protein monomers assemble into donut-shaped oligomers containing seven or eight protein monomers and rod-like structures. When bound to single-stranded DNA, RecT forms highly twisted nucleoprotein filaments that are 18 nm in diameter and have a helical pitch of 10 nm. When added to linear duplex DNA in the presence of active RecE protein (exonuclease VIII), filamentous nucleoprotein complexes are formed on the DNA ends and the DNA molecules are frequently cyclized through protein-protein interactions. PMID- 7490756 TI - Analysis of deletions induced in the genome of mammalian cells by ionizing radiation. AB - A theory is presented for the distribution in size of deletions induced by ionizing radiation, based on three assumptions: (1) deletions that are observed delete part or all of a gene to make a mutation, but not adjacent DNA sequences essential for survival of the mutant; (2) deletions are distributed at random along the DNA; (3) the probability of formation is proportional to the rate at which the two endpoints, which must meet to form the deletion, collide with each other. Experimental data for radiation-induced deletions in human and hamster hprt genes are in good agreement with calculations that assume the inducing lesion does not break the intracellular chromatin fiber; calculations assuming the inducing lesion is a break are not a good fit to the data. The low frequency of deletions observed in the hamster aprt gene is shown to be a consequence of the small gene size and the presence of a nearby essential DNA sequence, ensuring that most deletions affecting the gene also delete the essential sequence and are thus not observed. PMID- 7490757 TI - Determination of sequence specificity between a plasmid replication initiator protein and the origin of replication. AB - Staphylococcal plasmids of the pT181 family replicate by a rolling circle mechanism, requiring the activities of a plasmid-specified Rep protein. The initiation event involves site-specific phosphodiester bond cleavage by Rep within the replication origin, ori. In vitro the Rep proteins also display type-I topoisomerase activity specific for this plasmid family. Although the single site of bond cleavage, ICR II, is conserved among all members of the pT181 family, the plasmid-specific Rep proteins are able to discriminate between family members in vivo, initiating replication only from the cognate origin. The basis of such specificity is believed to be due to a non-covalent binding interaction between Rep and a DNA sequence adjacent to the site of phosphodiester bond cleavage. Using the RepD protein specified by plasmid pC221, we present data for the physical parameters of RepD:oriD complex formation. Quantification of the relative strengths of the non-covalent interactions for different but related ori target sequences, measured by gel mobility shift experiments, has yielded data that are in accord with the known specificity of the protein in vivo. Oligonucleotide competition experiments demonstrate that this interaction is indeed attributable to the specificity determinant, ICR III. Protein-DNA crosslinking methods show that a carboxyl-terminal proteolytic fragment of RepD makes a specific interaction with the ICR III region of its cognate replication origin. Analysis of topoisomerase rates indicates that the interaction between ICR III and the carboxyl terminus of the protein is required before a productive interaction, namely the phosphodiester bond cleavage at the ICR II, can occur. PMID- 7490758 TI - CDR walking mutagenesis for the affinity maturation of a potent human anti-HIV-1 antibody into the picomolar range. AB - We describe the investigation of methodologies for the creation of very high affinity human antibodies. The high affinity human antibody b4/12 was optimized for its affinity to the human envelope glycoprotein gp120 of human immunodeficiency virus type 1 (HIV-1). Five libraries of b4/12 were constructed by saturation mutagenesis of complementarity-determining regions (CDRs). Libraries of antibody Fab fragments were displayed on the surface of filamentous phage and selected in vitro for binding to immobilized gp120. Sequential and parallel optimization strategies of CDRs were examined. The sequential CDR walking strategy consistently yielded b4/12 variants of improved affinity in each of the four different optimization sequences examined. This resulted in a 96-fold improvement in affinity. Additivity effects in the antibody combining site were explored by combining independently optimized CDRs in the parallel optimization strategy. Six variants containing optimized CDRs were constructed. Improvement of affinity based on additivity effects proved to be unpredictable but did lead to a modest improvement in affinity. Indeed, only one of the six combinations demonstrated additivity. The highest affinity Fab prepared using this strategy was improved 420-fold in affinity. The affinity of this Fab was 15 pM as compared to 6.3 nM for b4/12. Examination of the kinetics of Fab binding to gp120 revealed that improvements in affinity were dominated by a slowing of the off-rate of the Fab. The methodology presented here provides a route for the improvement of the affinities of antibodies typical of tertiary immune responses into the picomolar range. Such improvements may have profound effects on the utility of antibodies as therapeutic and prophylactic agents. PMID- 7490759 TI - Lipid location in deoxycholate-treated purple membrane at 2.6 A. AB - A high resolution projection at 2.6 A of deoxycholate-treated purple membrane using only images has been obtained with a 200 keV FEG microscope operated at liquid helium temperature. Examination of this high quality map has allowed the following conclusions to be made: Comparison between the internal structure of the trimers of the native and the deoxycholate-treated crystal forms shows that almost every detail of the structure at high resolution is identical. The cell dimension change from 62.4 A to 57.9 A is accompanied by a loss of about half the normal lipids and a 2 degrees anticlockwise rotation of the trimer as a rigid body. Three of the lipids per bacteriorhodopsin molecule remain in identical positions relative to the trimer. In addition, from the projection map together with a packing analysis using the atomic model for bacteriorhodopsin, space for three further lipids has been identified making a total of six lipids per bacteriorhodopsin molecule in this crystal form. Finally, the surprisingly small rotation of the trimer between the two crystal forms with completely different Van der Waals contacts suggest that the crystals are held together by strong, long-range electrostatic interactions. PMID- 7490760 TI - The effect of temperature on the structure of vinblastine-induced polymers of purified tubulin: detection of a reversible conformational change. AB - Addition of the antimitotic drug vinblastine to solutions of purified tubulin induces the formation of helical polymers whose structure and type of aggregation is determined by the concentration of magnesium. While paracrystalline arrangements of single coils are observed at low concentrations of the ion, for concentrations higher than 6 mM free double-coiled spirals are obtained, which are indistinguishable from those obtained in the presence of microtubule associated proteins (MAPs). This result is consistent with a similar effect of magnesium and MAPs in neutralizing negative charges on the tubulin molecule and so allowing for lateral contacts between protofilaments. The effects that temperature has on the structure of both types of polymers, free spirals or paracrystals, have been monitored using time-resolved X-ray solution scattering. This study shows that a temperature increase: (1) affects the length and lateral aggregation of the spirals in the paracrystalline sample; (2) induces a reversible increase of the helical pitch in both types of polymers that closely follows the temperature change; (3) produces an irreversible aggregation of some of the protein in both types of polymers; and (4) can induce a reversible transformation from one type of structure to the other when the concentration of Mg2+ is in the boundary between the two ranges. We suggest that the changes in pitch are due to a temperature-induced conformational change of the tubulin molecule. This effect may be related to the structural modifications that result in the temperature-induced assembly of microtubules in vitro under normal conditions of assembly. PMID- 7490761 TI - Refined crystal structures of unligated adenylosuccinate synthetase from Escherichia coli. AB - Crystal structures of unligated adenylosuccinate synthetase from Escherichia coli in space groups P2(1) and P2(1)2(1)2(1) have been refined to R-factors of 0.199 and 0.206 against data to 2.0 and 2.5 A, respectively. Bond lengths and angles deviate from expected values by 0.011 A and 1.7 degrees for the P2(1) crystal form and by 0.015 A and 1.7 degrees for the P2(1)2(1)2(1) crystal form. The fold of the polypeptide chain is dominated by a central beta-sheet, which is composed of nine parallel strands and a tenth antiparallel strand. Extending off from this central beta-sheet are four subdomains. The four subdomains contribute loops of residues that are disordered or have high thermal parameters. At least three of these loops (residues 42 to 52, 120 to 131 and 298 to 304) contribute essential residues to the putative active site of the synthetase. In the absence of ligands, much of the active site of the synthetase exists in an ill-defined conformational state. Two, nearly independent regions contribute residues to the interface between polypeptide chains of the synthetase dimer. A pair of helices (H4 and H5) interact with their symmetry-equivalent mates by way of residues that are not conserved amongst the known sequences of the synthetase. The second interface region involves conserved residues belonging to structural elements that connect strands of the central beta-sheet. Residues putatively involved in the binding of IMP lie at or near the interface between polypeptide chains of the dimer. Of the four sequence elements putatively common to all GTP hydrolases, the synthetase has only the guanine recognition element and a glycine-rich loop (P loop). Although the base recognition element is essentially identical with those of the p21 ras and G alpha proteins, the P-loop of the synthetase is extended in size relative to the P-loops of other GTP hydrolases. The P-loop has two acid residues (Asp13 and Glu14), which are found in the P-loops of only the synthetase family. Glu14 may be involved in the stabilization of the enlarged P-loop of the synthetase, whereas Asp13 may play a role in catalysis and in the coordination of Mg2+. The structural elements of the p21 ras and G alpha proteins responsible for binding Mg2+ are either absent from the synthetase or unavailable for the coordination of metal cations. PMID- 7490762 TI - Bacillus thuringiensis CryIA(a) insecticidal toxin: crystal structure and channel formation. AB - The activated 65 kDa lepidopteran-specific CryIA(a) toxin from the commercially most important strain Bacillus thuringiensis var. kurstaki HD-1 has been investigated by X-ray diffraction and for its ability to form channels in planar lipid bilayers. Its three-dimensional structure has been determined by a multiple isomorphous replacement method and refined at 2.25 A resolution to an R-factor of 0.168 for data with I > 2 delta (I). The toxin is made of three distinct domains. The N-terminal domain is a bundle of eight alpha-helices with the central, relatively hydrophobic helix surrounded by amphipathic helices. The middle and C terminal domains contain mostly beta-sheets. Comparison with the structure of CryIIIA, a coleopteran-specific toxin, shows that although the fold of these two proteins is similar, there are significant structural differences within domain II. This finding supports the conclusions from genetic studies that domain II is involved in recognition and binding to cell surface receptors. The distribution of electrostatic potential on the surface of the molecule is non-uniform and identifies one side of the alpha-helical domain as negatively charged. The predominance of arginine residues as basic residues ensures that the observed positive charge distribution is also maintained in the highly alkaline environment found in the lepidopteran midgut. Structurally important salt bridges that are conserved across Cry sequences were identified and their possible role in toxin action was postulated. In planar lipid bilayers, CryIA(a) forms cation selective channels, whose conductance is significantly smaller than that reported for CryIIIA but similar to those of other Cry toxins. PMID- 7490763 TI - Automated assignment of simulated and experimental NOESY spectra of proteins by feedback filtering and self-correcting distance geometry. AB - A new method for automatically assigning proton-proton NOESY spectra is described and demonstrated for simulated and experimental spectra of the proteins dendrotoxin K, alpha-amylase inhibitor tendamistat and the DNA-binding domain of the 434 repressor protein. The method assigns the NOESY spectrum and calculates three-dimensional protein structures simultaneously, using a list of proton chemical shifts and 3JNH alpha coupling constants. An ensemble of structures is iteratively calculated by self-correcting distance geometry from unambiguous and selected ambiguous NOESY cross peaks. New structure based filters recognize the correct constraints from the ambiguous cross peak list. For the first round of assignment neither a preliminary initial structure nor a sufficient set of unambiguous NOESY cross peaks is needed. The method can also be applied to cross peak lists containing hundreds of noise peaks. For an assumed tolerance of +/- 0.01 ppm in the chemical shifts of the peak positions, only about 10% of the NOESY cross peaks can be unambiguously assigned based on their chemical shifts alone. Our automated method assigned about 80% of all cross peaks with this chemical shift tolerance, and 95 to 99% of the assignments were correct. The average pairwise RMSD for the backbone atoms of the ten best final structures is about 1.5 A in all three proteins and the previously determined NMR solution structures are always embedded in this structure bundle. We regard our method as a highly practical tool for automatic calculation of three-dimensional protein structures from NMR spectra with minimal human interference. PMID- 7490764 TI - Mechanism of protein stabilization by disulfide bridges: calorimetric unfolding studies on disulfide-deficient mutants of the alpha-amylase inhibitor tendamistat. AB - The present differential scanning calorimetry and circular dichroism studies on the mechanism of protein stabilization by disulfide bonds were concerned with two questions: is the increase in unfolding entropy upon removal of disulfide links sufficient for the explantation of the general stability decrease of disulfide deficient mutants? Is it immaterial by which residue cysteine residues are replaced when disulfide bridges are to be opened? To answer these questions we investigated two disulfide bridge mutants of the alpha-amylase inhibitor Tendamistat where the large loop (C45A/C73A) or the small loop (C11A/C27A) had been opened by recombinant DNA techniques, and we compared the stability of the mutated proteins with that of wild-type Tendamistat published previously. To elucidate the significance of the nature of the group that replaces Cys we introduced in position 27 of the small loop four different amino acids instead of Cys: Ala, Leu, Ser and Thr. Surprisingly, opening of the small loop (17 residues) causes larger destabilization than opening of the large loop comprising 29 residues. The thermodynamic parameters at pH 7.0 are: wild-type: t1/2 = 81.6 degrees C, delta Hcal = 296 kJ mol-1, large loop mutant (C45A/C73A): t1/2 = 58.6 degrees C, delta Hcal = 225 kJ mol-1 and small loop mutant (C11A/C27A): t1/2 = 42.7 degrees C, delta Hcal = 135 kJ mol-1. This finding is at variance with the entropy hypothesis. The relative contributions to stability of enthalpic and entropic terms can be varied by a proper choice of substitutions. While the destabilization originating from C45A/C73A exchanges in the large loop turns out to be purely entropic, the stability decreases of the small loop mutants are caused by changes in both enthalpic and entropic terms. Leu or Ser in position 27 leads to an overall enthalpic destabilization. Thr in position 27 increases the transition enthalpy of this mutant to the value of the wild-type protein but increases at the same time the value of the transition entropy with the result of an overall entropic destabilization. Finally, in the C11A/C27A small loop mutant of lowest stability a very large enthalpic destabilization occurs, which is, however, partly counterbalanced by a reduction in the transition entropy. The preferential perturbation of the native state by the mutations is manifest in the increase of the native state heat capacity relative to that of the wild-type protein and the identity of the heat capacity of the unfolded state.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 7490765 TI - Canonical structure repertoire of the antigen-binding site of immunoglobulins suggests strong geometrical restrictions associated to the mechanism of immune recognition. AB - Is the structural repertoire of immunoglobulins free to adopt an almost infinite number of conformations to build the diversity of the immune response or does it take advantage of only a few conformations? In this paper we study this question by applying the canonical structure model to characterize the structural repertoire of immunoglobulins. The results found, indicate that only ten combinations out of the 300 possible different canonical structure classes (combinations of canonical structures), make up 87% of 381 sequences analyzed. This suggests that the structural repertoire of immunoglobulins is restricted to the preferential use of a small number of canonical structure classes. The possible functional significance of these results was studied by analyzing the correspondence between the observed canonical structural repertoire implicit in Ig sequences and the types of antigens recognized. Two different sets of canonical structure classes were distinguished: one with preference for some specific types of antigens like proteins, polysaccharides or haptens, and the other with multi-specific binding capabilities. Analysis of antibodies of known three-dimensional structure shows that for two specific classes, the canonical conformations of H2 and L1 determine the geometrical characteristics of the antigen-binding site, while at least in one multi-specific class, the changes in the general geometry of the antigen-binding site are produced by different conformations of H3. Implications of these results for the molecular recognition process mediated by immunoglobulins are discussed. PMID- 7490766 TI - Preferential heterodimeric parallel coiled-coil formation by synthetic Max and c Myc leucine zippers: a description of putative electrostatic interactions responsible for the specificity of heterodimerization. AB - The oncoprotein c-Myc must heterodimerize with Max to bind DNA and perform its oncogenic activity. The c-Myc-Max heterodimer binds DNA through a basic helix loop-helix leucine zipper (b-HLH-zip) motif and it is proposed that leucine zipper domains could, in concert with the HLH regions, provide the specificity and stability of the b-HLH-zip motif. In this context, we have synthesized the peptides corresponding to the leucine zipper domains of Max and c-Myc with a N terminal Cys-Gly-Gly linker and studied their dimerization behavior using reversed-phase HPLC and CD spectroscopy. The preferential formation of a fully helical parallel c-Myc-Max heterodimeric coiled-coil was observed under air oxidation and redox conditions at neutral pH. We show that the stability and the helicity of the disulfide-linked c-Myc-Max heterostranded coiled-coil is modulated by pH, with a maximum around pH 4.5, supporting the existence of stabilizing and specific interhelical electrostatic interactions. We present a molecular model of the c-Myc-Max heterostranded coiled-coil describing potential electrostatic interactions responsible for the specificity of the interaction, the main feature being putative buried electrostatic interactions between a histidine side-chain (in the Max leucine zipper) and two glutamic acid side chains (in the c-Myc leucine zipper) at the heterodimer interface. This model is supported by the fact that the apparent pKa (as determined by [1H]-NMR spectroscopy) of this histidine side-chain at 25 degrees C is 0.42 (+/- 0.05) pKa units higher in the folded form than in the unfolded form. This indicates that the charged histidine side-chain contributes approximately 0.57 (+/- 0.07) kcal/mol (2.38 (+/- 0.30) kJ/mol) of stabilization free energy to the c-Myc-Max heterostranded coiled-coil through favorable electrostatic interaction. PMID- 7490768 TI - The evolution of protamine P1 genes in dasyurid marsupials. AB - We report the complete DNA sequences of the protamine P1 gene and flanking regions for 13 species of the marsupial family Dasyuridae. The structure of the protamine locus is conserved in dasyurids and consists of two exons (of lengths 142-151 and 47 bp) separated by an intron (208-240 bp). A key feature of the dasyurid intron is a 38-40 bp duplication found in all species examined to date. This duplication apparently predates the radiation of modern dasyurid lineages and may be homologous to a similar feature in the marsupial mole (Notoryctes). Sequences from a species of Planigale demonstrate that this genus is unique among marsupials in possessing cysteine residues in its protamine P1 molecules. Cysteines may provide enhanced chemical stability for condensed sperm nuclei, a physiological feature that would converge on the common eutherian pattern. Phylogenetic analysis of the protamine genes yields a tree that is largely congruent with previous molecular systematic studies in two areas: (1) There are three main dasyurid lineages corresponding to the Sminthopsinae. Dasyurinae, and Phascogalinae; (2) Dasyurinae and Phascogalinae are sister groups. This study is the first estimate of dasyurid relationships based on a nuclear DNA sequence. PMID- 7490767 TI - The evolution of the type I interferon gene family in mammals. AB - A phylogenetic analysis of mammalian type I interferon (IFN) genes showed: (1) that the three main subfamilies of these genes in mammals (IFN-beta, IFN-alpha, and IFN-omega) diverged after the divergence of birds and mammals but before radiation of the eutherian orders and (2) that IFN-beta diverged first. Although apparent cases of interlocus recombination among mouse IFN-alpha genes were identified, the hypothesis that coding regions of IFN-alpha genes have been homogenized within species by interlocus recombination was not supported. Flanking regions as well as coding regions of IFN-alpha were more similar within human and mouse than between these species; and reconstruction of the pattern of nucleotide substitution in IFN-alpha coding regions of four mammalian species by the maximum parsimony method suggested that parallel substitutions have occurred far more frequently between species than within species. Therefore, it seems likely that IFN-alpha genes have duplicated independently within different eutherian orders. In general, type I IFN genes are subject to purifying selection, which in the case of IFN-alpha and IFN-beta is strongest in the putative receptor-binding domains. However, analysis of the pattern of nucleotide substitution among IFN-omega genes suggested that positive Darwinian selection may have acted in some cases to diversify members of this subfamily at the amino acid level. PMID- 7490769 TI - Concerted evolution in protists: recent homogenization of a polyubiquitin gene in Trichomonas vaginalis. AB - Ubiquitin is a 76-amino-acid protein with a remarkably high degree of conservation between all known sequences. Ubiquitin genes are almost always multicopy in eukaryotes, and often are found as polyubiquitin genes--fused tandem repeats which are coexpressed. Seventeen ubiquitin sequences from the amitochondrial protist Trichomonas vaginalis have been examined here, including an 11-repeat fragment of a polyubiquitin gene. These sequences reveal a number of interesting features that are not seen in other eukaryotes. The predicted amino acid sequences lack several universally conserved residues, and individual units do not always encode identical peptides as is usually the case. On the nucleotide level, these repeats are in general highly variable, but one region in the polyubiquitin is extremely homogeneous, with seven repeats absolutely identical. Such extended stretches of homogeneity have never been observed in ubiquitin genes and since substitutions are common in other coding units, it is likely that these repeats are the product of a very recent homogenization or amplification. PMID- 7490770 TI - Fungal origin by horizontal transfer of a plant mitochondrial group I intron in the chimeric CoxI gene of Peperomia. AB - We present phylogenetic evidence that a group I intron in an angiosperm mitochondrial gene arose recently by horizontal transfer from a fungal donor species. A 1,716-bp fragment of the mitochondrial coxI gene from the angiosperm Peperomia polybotrya was amplified via the polymerase chain reaction and sequenced. Comparison to other coxI genes revealed a 966-bp group I intron, which, based on homology with the related yeast coxI intron aI4, potentially encodes a 279-amino-acid site-specific DNA endonuclease. This intron, which is believed to function as a ribozyme during its own splicing, is not present in any of 19 coxI genes examined from other diverse vascular plant species. Phylogenetic analysis of intron origin was carried out using three different tree-generating algorithms, and on a variety of nucleotide and amino acid data sets from the intron and its flanking exon sequences. These analyses show that the Peperomia coxI gene intron and exon sequences are of fundamentally different evolutionary origin. The Peperomia intron is more closely related to several fungal mitochondrial introns, two of which are located at identical positions in coxI, than to identically located coxI introns from the land plant Marchantia and the green alga Prototheca. Conversely, the exon sequence of this gene is, as expected, most closely related to other angiosperm coxI genes. These results, together with evidence suggestive of co-conversion of exonic markers immediately flanking the intron insertion site, lead us to conclude that the Peperomia coxI intron probably arose by horizontal transfer from a fungal donor, using the double-strand-break repair pathway. The donor species may have been one of the symbiotic mycorrhizal fungi that live in close obligate association with most plants. PMID- 7490771 TI - Relative roles of primary sequence and (G + C)% in determining the hierarchy of frequencies of complementary trinucleotide pairs in DNAs of different species. AB - To an approximation Chargaff's rule (%A = %T; %G = %C) applies to single-stranded DNA. In long sequences, not only complementary bases but also complementary oligonucleotides are present in approximately equal frequencies. This applies to all species studied. However, species usually differ in base composition. With the goal of understanding the evolutionary forces involved, I have compared the frequencies of trinucleotides in long sequences and their shuffled counterparts. Among the 32 complementary trinucleotide pairs there is a hierarchy of frequencies which is influenced both by base composition (not affected by shuffling the order of the bases) and by base order (affected by shuffling). The influence of base order is greatest in DNA of 50% G + C and seems to reflect a more fundamental hierarchy of dinucleotide frequencies. Thus if TpA is at low frequency, all eight TpA-containing trinucleotides are at low frequency. Mammals and their viruses share similar hierarchies, with intra- and intergenomic differences being mainly associated with differences in base composition (percentage G + C). E. coli and, to a lesser extent, Drosophila melanogaster hierarchies differ from mammalian hierarchies; this is associated with differences both in base composition and in base order. It is proposed that Chargaff's rule applies to single-stranded DNA because there has been an evolutionary selection pressure favoring mutations that generate complementary oligonucleotides in close proximity, thus creating a potential to form stem loops. These are dispersed throughout genomes and are rate-limiting in recombination. Differences in (G + C)% between species would impair interspecies recombination by interfering with stem-loop interactions. PMID- 7490772 TI - Sense in antisense? AB - A correspondence between open reading frames in sense and antisense strands is expected from the hypothesis that the prototypic triplet code was of general form RNY, where R is a purine base, N is any base, and Y is a pyrimidine. A deficit of stop codons in the antisense strand (and thus long open reading frames) is predicted for organisms with high G + C percentages; however, two bacteria (Azotobacter vinelandii, Rhodobacter capsulatum) have larger average antisense strand open reading frames than predicted from (G + C)%. The similar codon frequencies found in sense and antisense strands can be attributed to the wide distribution of inverted repeats (stem-loop potential) in natural DNA sequences. PMID- 7490773 TI - Molecular evolution of the hepatitis B virus genome. AB - The hepatitis B virus (HBV) has a circular DNA genome of about 3,200 base pairs. Economical use of the genome with overlapping reading frames may have led to severe constraints on nucleotide substitutions along the genome and to highly variable rates of substitution among nucleotide sites. Nucleotide sequences from 13 complete HBV genomes were compared to examine such variability of substitution rates among sites and to examine the phylogenetic relationships among the HBV variants. The maximum likelihood method was employed to fit models of DNA sequence evolution that can account for the complexity of the pattern of nucleotide substitution. Comparison of the models suggests that the rates of substitution are different in different genes and codon positions; for example, the third codon position changes at a rate over ten times higher than the second position. Furthermore, substantial variation of substitution rates was detected even after the effects of genes and codon positions were corrected; that is, rates are different at different sites of the same gene or at the same codon position. Such rates after the correction were also found to be positively correlated at adjacent sites, which indicated the existence of conserved and variable domains in the proteins encoded by the viral genome. A multiparameter model validates the earlier finding that the variation in nucleotide conservation is not random around the HBV genome. The test for the existence of a molecular clock suggests that substitution rates are more or less constant among lineages. The phylogenetic relationships among the viral variants were examined. Although the data do not seem to contain sufficient information to resolve the details of the phylogeny, it appears quite certain that the serotypes of the viral variants do not reflect their genetic relatedness. PMID- 7490774 TI - Neighboring base composition is strongly correlated with base substitution bias in a region of the chloroplast genome. AB - Nucleotide sequence from a region of the chloroplast genome is presented for 12 species spanning four subfamilies of the grass family. The region contains the coding sequence for the rbcL gene and the intergenic spacer between the gene coding the large subunit of ribulose-1,5-bisphosphate carboxylase/oxygenase (rbcL) and the photosystem I gene psaI. This intergenic spacer contains a pseudogene for rpl23 as well as two noncoding segments with different A+T contents. Using the sequence of rbcL a chloroplast phylogeny of this family was constructed by parsimony. Variable sites of the two noncoding segments were traced onto the phylogeny to study the dynamics of base substitution. This was also performed for the fourfold-degenerate sites of the rbcL gene. A wide variation in transversion/transition is observed between the two noncoding segments and between the noncoding DNA and the fourfold-degenerate sites of rbcL. This variation is correlated with regional A+T content. As regional A+T content decreases, the ratio of transversions to transitions also decreases. Substitutions were then scored in relation to neighboring base composition. The composition of the two bases immediately flanking each substitution is highly correlated with the transversion/transition bias. When both the 5' and 3' flanking bases are an A or a T, transversions are observed 2.2 times as frequently as transitions. When either or both neighbors are a C or a G, the opposite trend is found; transitions are observed 1.5 times more frequently than transversions. PMID- 7490776 TI - Structural evolution of the Drosophila 5S ribosomal genes. AB - We compare the 5S gene structure from nine Drosophila species. New sequence data (5S genes of D. melanogaster, D. mauritiana, D. sechellia, D. yakuba, D. erecta, D. orena, and D. takahashii) and already-published data (5S genes of D. melanogaster, D. simulans, and D. teissieri) are used in these comparisons. We show that four regions within the Drosophila 5S genes display distinct rates of evolution: the coding region (120 bp), the 5'-flanking region (54-55 bp), the 3' flanking region (21-22 bp), and the internal spacer (149-206 bp). Intra- and interspecific heterogeneity is due mainly to insertions and deletions of 6-17-bp oligomers. These small rearrangements could be generated by fork slippages during replication and could produce rapid sequence divergence in a limited number of steps. PMID- 7490775 TI - The pvB370 BamHI satellite DNA family of the Drosophila virilis group and its evolutionary relation to mobile dispersed genetic pDv elements. AB - The pvB370 BamHI tandemly repeated satellite DNA family was isolated and analyzed in eight species of the Drosophila virilis group and is probably common to all its members. Unexpectedly, the satellite DNA family showed similar levels of intra- and interspecific sequence variability. An EMBL gene-bank search revealed a high degree of similarity between the members of the pvB370 BamHI satellite DNA sequence family and the direct terminal repeats of the mobile dispersed genetic pDv elements described in D. virilis and other species of the group. However, no similarity was observed to the transcribed and translated part of the pDv elements. It is suggested that the satellite DNA repeats are phylogenetically older than the mobile dispersed genetic pDv element and that the functional pDv elements might have derived from the satellite DNA family through an insertion of a tandemly repeated 36-bp transcription unit. PMID- 7490777 TI - Molecular evolution of the mitochondrial 12S rRNA in Ungulata (mammalia). AB - The complete 12S rRNA gene has been sequenced in 4 Ungulata (hoofed eutherians) and 1 marsupial and compared to 38 available mammalian sequences in order to investigate the molecular evolution of the mitochondrial small-subunit ribosomal RNA molecule. Ungulata were represented by one artiodactyl (the collared peccary, Tayassu tajacu, suborder Suiformes), two perissodactyls (the Grevy's zebra, Equus grevyi, suborder Hippomorpha; the white rhinoceros, Ceratotherium simum, suborder Ceratomorpha), and one hyracoid (the tree hyrax, Dendrohyrax dorsalis). The fifth species was a marsupial, the eastern gray kangaroo (Macropus giganteus). Several transition/transversion biases characterized the pattern of changes between mammalian 12S rRNA molecules. A bias toward transitions was found among 12S rRNA sequences of Ungulata, illustrating the general bias exhibited by ribosomal and protein-encoding genes of the mitochondrial genome. The derivation of a mammalian 12S rRNA secondary structure model from the comparison of 43 eutherian and marsupial sequences evidenced a pronounced bias against transversions in stems. Moreover, transversional compensatory changes were rare events within double stranded regions of the ribosomal RNA. Evolutionary characteristics of the 12S rRNA were compared with those of the nuclear 18S and 28S rRNAs. From a phylogenetic point of view, transitions, transversions and indels in stems as well as transversional and indels events in loops gave congruent results for comparisons within orders. Some compensatory changes in double-stranded regions and some indels in single-stranded regions also constituted diagnostic events. The 12S rRNA molecule confirmed the monophyly of infraorder Pecora and order Cetacea and demonstrated the monophyly of the suborder Ruminantia was not supported and the branching pattern between Cetacea and the artiodacytyl suborders Ruminantia and Suiformes was not established. The monophyly of the order Perissodactyla was evidenced, but the relationships between Artiodactyla, Cetacea, and Perissodactyla remained unresolved. Nevertheless, we found no support for a Perissodactyla + Hyracoidea clade, neither with distance approach, nor with parsimony reconstruction. The 12S rRNA was useful to solve intraordinal relationships among Ungulata, but it seemed to harbor too few informative positions to decipher the bushlike radiation of some Ungulata orders, an event which has most probably occurred in a short span of time between 55 and 70 MYA. PMID- 7490778 TI - Revised dinoflagellate phylogeny inferred from molecular analysis of large subunit ribosomal RNA gene sequences. AB - The nucleotide sequence analysis of the PCR products corresponding to the variable large-subunit rRNA domains D1, D2, D9, and D10 from ten representative dinoflagellate species is reported. Species were selected among the main laboratory-grown dinoflagellate groups: Prorocentrales, Gymnodiniales, and Peridiniales which comprise a variety of morphological and ecological characteristics. The sequence alignments comprising up to 1,000 nucleotides from all ten species were employed to analyze the phylogenetic relationships among these dinoflagellates. Maximum parsimony and neighbor-joining trees were inferred from the data generated and subsequently tested by bootstrapping. Both the D1/D2 and the D9/D10 regions led to coherent trees in which the main class of dinoflagellates. Dinophyceae, is divided in three groups: prorocentroid, gymnodinioid, and peridinioid. An interesting outcome from the molecular phylogeny obtained was the uncertain emergence of Prorocentrum lima. The molecular results reported agreed with morphological classifications within Peridiniales but not with those of Prorocentrales and Gymnodiniales. Additionally, the sequence comparison analysis provided strong evidence to suggest that Alexandrium minutum and Alexandrium lusitanicum were synonymous species given the identical sequence they shared. Moreover, clone Gg1V, which was determined Gymnodinium catenatum based on morphological criteria, would correspond to a new species of the genus Gymnodinium as its sequence clearly differed from that obtained in G. catenatum. The sequence of the amplified fragments was demonstrated to be a valuable tool for phylogenetic and taxonomical analysis among these highly diversified species. PMID- 7490781 TI - Intravenous reproductive and developmental toxicity studies of cimadronate (YM175), a novel bisphosphonate, in rats and rabbits. AB - Cimadronate (YM175) is a novel bisphosphonate with potent inhibitory activity on bone resorption under development for the treatment of tumor-induced hypercalcemia, metastatic bone disease and osteoporosis. We conducted intravenous reproductive toxicity and teratology studies (Segment I, II and III) of this compound in rats and teratology study in rabbits. The test compound was dissolved in physiological saline, which was also given as the vehicle control. Rats were administered at a dosage of 0.06, 0.16 and 0.62 mg/kg/day in the male Segment I study. Dose levels in the other studies in rats including the female Segment I were 0.16, 0.31 and 0.62 mg/kg/day. In the Segment I study, no treatment-related abnormalities were observed in reproductive parameters or fetuses. In the Segment II study, slightly retarded fetal ossification was noted at 0.31 mg/kg/day or more, but the incidence of malformation did not increase. In the Segment III study, death of the dams and abnormal tooth growth of offspring were observed at 0.16 mg/kg/day or more. Further Segment III study showed that the no toxic effect level was 0.003 mg/kg/day. In the rabbit teratology study, dose levels were 0.01, 0.025 or 0.05 mg/kg/day. No toxic effects on pregnant females or their litters were observed at up to 0.05 mg/kg/day. PMID- 7490779 TI - Goldfish cones secrete a two-repeat interphotoreceptor retinoid-binding protein. AB - Vitamin A and fatty acids are critical to photoreceptor structure, function, and development. The transport of these nutrients between the pigment epithelium and neural retina is mediated by interphotoreceptor retinoid-binding protein (IRBP). IRBP, a 133-kDa (human) glycolipoprotein, is the major protein component of the extracellular matrix separating these two cell layers. In amphibians and mammals, IRBP consists of four homologous repeats of about 300 amino acids which form two retinol and four fatty acid-binding sites. Here we show that IRBP in teleosts is a simpler protein composed of only two repeats. Western blot analysis shows that goldfish IRBP is half the size (70 kDa) of IRBP in higher vertebrates. Metabolic labeling studies employing Brefeldin A taken together with in situ hybridization studies and the presence of a signal peptide show that goldfish IRBP is secreted by the cone photoreceptors. The translated amino acid sequence has a calculated molecular weight of 66.7 kDa. The primary structure consists of only two homologous repeats with a similarity score of 52.5%. The last repeats of human and goldfish IRBPs are 69.1% similar with hydrophobic regions being the most similar. These data suggest that two repeats were lost during the evolution of the ray-finned fish (Actinopterygii), or that the IRBP gene duplicated between the emergence of bony fish (Osteichthyes) and amphibians. Acquisition of a multirepeat structure may reflect evolutionary pressure to efficiently transport higher levels of hydrophobic molecules within a finite space. Quadruplication of an ancestral IRBP gene may have been an important event in the evolution of photo receptors in higher vertebrates. PMID- 7490782 TI - [Reproductive toxicity of iodixanol, a new non-ionic, iso-tonic contrast medium in rats and rabbits]. AB - The reproductive toxicity of iodixanol, a non-ionic isotonic contrast medium, was investigated in rats and rabbits. Iodixanol was administered intravenously to male rats at doses of 0.3, 1.0 or 2.0 gI/kg/day for 80 days or more until successful copulation. Female rats were treated with iodixanol at the same dose levels for at least 14 days prior to and in the early stage of pregnancy. No remarkable adverse effects on fertility were noted at any dose level. Increased water consumption and swollen paws/face were observed among male rats receiving > 1.0 gI/kg/day. In addition, decreased food consumption was observed in males of the 2.0 gI/kg/day group. In female rats, no remarkable adverse effects were noted at any dose level. Iodixanol elicited no evidence of teratogenicity when administered during the fetal organogenesis period to pregnant rats and rabbits at doses of 0.3, 1.0 or 2.0 gI/kg/day. No remarkable changes were observed in rat or rabbit fetuses, nor in dams of any dose group. In a perinatal and postnatal dosing study in rats using doses of 0.3, 1.0 or 2.0 gI/kg/day, iodixanol did not adversely affect the reproductive performance of the treated parent animals and pre- and postnatal development of their offspring. PMID- 7490780 TI - Molecular phylogeny of Allomyces macrogynus: congruency between nuclear ribosomal RNA- and mitochondrial protein-based trees. AB - We have sequenced the nuclear and mitochondrial small subunit rRNA genes (rns) and the mitochondrial genes coding for subunits 1 and 3 of the cytochrome oxidase (cox1 and cox3, respectively) of the chytridiomycete Allomyces macrogynus. Phylogenetic trees inferred from the derived COX1 and COX3 proteins and the nuclear rns sequences show with good bootstrap support that A. macrogynus is an early diverging fungus. The trees inferred from mitochondrial rns sequences do not yield a topology that is supported by bootstrap analysis. The similarity and the relative robustness of the nuclear rns and the mitochondrial protein-derived phylogenetic trees suggest that protein sequences are of higher value than rRNA sequences for reconstructing mitochondrial evolution. In addition, our trees support a monophyletic origin of mitochondria for the range of analyzed eukaryotes. PMID- 7490783 TI - [Antigenicity study of iodixanol, a new non-ionic contrast medium]. AB - Antigenic potential of iodixanol, a new non-ionic contrast agent, was evaluated by systemic anaphylaxis (SA) test and homologous passive cutaneous anaphylaxis (Homo-PCA) test using guinea pigs and IgE mediated heterologous PCA (Hetero-PCA) test using rats passively sensitized with mouse sera. SA test was conducted in guinea pigs sensitized with iodixanol at dose levels of 32 or 320 mgI/kg emulsified with Freund's complete adjuvant, followed by challenge with iodixanol (320 mgI/kg). Four- or 24-hr Homo-PCA test was performed with sera from the sensitized animals. Neither positive SA nor Homo-PCA reaction was observed. Hetero-PCA test was performed with the sera from the mice which were sensitized with iodixanol (32 or 320 mgI/kg) with aluminum hydroxide gel. No positive Hetero PCA reaction was detected in the sera, followed by challenge with iodixanol (320 mgI/kg). Thus, it is concluded that iodixanol possesses no antigenic potential under the present experimental conditions. PMID- 7490784 TI - [Mutagenicity studies of iodixanol, a new non-ionic isotonic contrast medium]. AB - A new non-ionic isotonic contrast medium, iodixanol (a diastereomeric mixture of 5,5'-[(2-hydroxytrimethylene) bis (acetylimino)] bis [N, N'-bis (2,3 dihydroxypropyl)-2,4,6-triiodo-1,3-benzene-dicarboxamide]) was studied for mutagenicity by using the Ames method, in vitro cytogenetics and micronucleus test. Iodixanol had no mutagenic effect on S. typhimurium (TA1535, TA1537, TA1538, TA98 and TA100) or E. coli (WP2uvrA) in the reverse mutation assay with or without metabolic activations. In the cytogenetic study, iodixanol had no effect on the Chinese hamster cells with or without metabolic activations. Intravenous injections of iodixanol for two times at a dose of 800, 1,600 or 3,200 mgI/kg in the mouse micronucleus test did not increase the incidence of micronucleated polychromatic erythrocytes. These results show that iodixanol has no mutagenic potential. PMID- 7490785 TI - [General pharmacological study of iodixanol, a new non-ionic isotonic contrast medium]. AB - The general pharmacological study of iodixanol, a non-ionic isotonic contrast medium, was conducted. 1) Iodixanol administered intravenously over a dose range of 320 to 3,200 mgI/kg had little or no effect on the general behavior, spontaneous locomotor activity, hexobarbital sleeping time, pain response, electroshock- or pentylenetetrazol-induced convulsion (mouse), EEG or body temperature (rabbit), gastrointestinal propulsion (mouse) or skeletal muscle contraction (rabbit). Iodixanol had no specific interaction with acetylcholine, histamine, serotonin, nicotin, BaCl2 (ileum), methacholine (trachea), isoprenaline (atrium) or oxytocin (pregnant uterus), nor had any effect on spontaneous contractility (atrium and uterus), or transmural electrostimulation induced contractility (vas deferens) at concentrations of < or = 3.2 x 10(-3) gI/ml in vitro. Iodixanol had no effect on the cardiovascular system of dog, except that it increased femoral blood flow and respiratory rate at doses of > or = 1,000 mgI/kg. Iodixanol at 3,200 mgI/kg i.v. reduced urine output with a decrease in Na+ and Cl- excretion, whereas at 320 mgI/kg i.v., it slightly increased urine output (rat). 2) Injections of iodixanol into the cerebroventricular (0.96, 9.6 mgI/mouse and 3.2, 32 mgI/rat), left ventricular (1,920, 6,400 mgI/dog) or coronary artery (640, 1,920 mgI/dog) had no conspicuous effect on the central nervous system or the cardiovascular system, respectively. There was no marked difference among iodixanol, iohexol and iopamidol in this respect. Vascular pain during injection into the femoral artery (300-320 mgI/guinea pig) appeared to be less intense with iodixanol, compared with the other contrast media iohexol and iopamidol. These results suggest that intravenous injection of iodixanol is relatively free from pharmacological activity, and effects of iodixanol on the central nervous system (intracerebroventricular injection) and cardiovascular system (intra-left ventricular and -coronary injections) are comparable to those of iohexol and iopamidol. Furthermore, intra-femoral injection of iodixanol has less of a tendency to produce vascular pain than those of iohexol and iopamidol. PMID- 7490787 TI - Intravenous single and repeated dose toxicity studies of cimadronate (YM175), a novel bisphosphonate, in beagle dogs. AB - Cimadronate (YM175) is a novel bisphosphonate with potent inhibitory activity on bone resorption under development for the treatment of tumor-induced hypercalcemia, metastatic bone disease and osteoporosis. We conducted intravenous single and repeated dose toxicity studies of cimadronate in beagle dogs. In the single dose study, animals received a single dose of 0.3, 1, 3 or 10 mg/kg of cimadronate and the animals were observed for at least 14 days. At 10 mg/kg, both the male and female dog showed toxic signs such as vomiting, decreased locomotor activities and hypothermia and were killed in extremis within a week after dosing. In the 30-day study, animals received cimadronate at a dosage of 0 (vehicle), 0.03, 0.1, 0.3 or 1 mg/kg/day. At 0.03 mg/kg/day or more, histological findings indicated an increased amount of primary spongiosa in the rib and ilium. At 1 mg/kg/day, degenerative nephropathy, aggregation of spermatozoa and glandular hypoplasia of the prostate gland were observed. On day 16 of dosing one male animal died of acute renal failure. In the 26-week study, animals received cimadronate once weekly at a dosage of 0 (vehicle), 0.31, 0.62, or 1.25 mg/kg. Histopathological examination showed an increased amount of primary spongiosa in the rib at all dosage levels. In addition, similar findings were observed in the lumbar vertebrae at 1.25 mg/kg/week. Histopathological changes in the kidney and male reproductive organs were not observed. PMID- 7490786 TI - Intravenous single and repeated dose toxicity studies of cimadronate (YM175), a novel bisphosphonate, in rats. AB - Cimadronate (YM175) is a novel bisphosphonate with potent inhibitory activity on bone resorption under development for the treatment of tumor-induced hypercalcemia, metastatic bone disease and osteoporosis. We conducted toxicity studies of cimadronate intravenously, single dose, 30-day repeated dose and 26 week weekly dose in F344 rats. In the single dose study, cimadronate was administered to rats and the animals were observed for 14 days. Major toxic symptoms were decreased motility and piloerection and LD50 values were 23 mg/kg for males and 21 mg/kg for females. In the 30-day study, the animals received cimadronate at doses of 0, 0.16, 0.31, 0.62 or 1.25 mg/kg/day. At 0.16 mg/kg/day or more, an increased amount of primary spongiosa was observed in the femur. At 0.62 mg/kg/day or more, renal and testicular/epididymal toxicity were observed. After a 30-day recovery period, the finding in the kidney disappeared, but the findings remained in the bone, testis and epididymis. In the 26-week weekly dose study, animals received cimadronate at doses of 0, 0.31, 0.62 and 1.25 mg/kg/week. At 0.31 mg/kg/week or more, an increased amount of primary spongiosa was seen in the femur. Renal and testicular/epididymal toxicity, however, were not observed. PMID- 7490788 TI - The mechanism of lactitol (NS-4) in inducing adrenomedullary proliferative lesion in rats. AB - We used 13-week repeated oral administration of lactitol as part of a study to clarify the mechanism by which lactitol induces the proliferation of adrenomedullary chromaffin cells. There was a marked increase in urinary calcium (Ca) excretion even though the lactitol administration had no effect on the blood Ca level. A tendency for an increase in adrenal venous blood epinephrine (EPI) and norepinephrine (NE) concentrations was seen. Organ weight measurement of adrenal glands revealed a tendency for an increase in absolute weight and a significant increase in relative weight. Morphometric analysis of adrenomedullary chromaffin cells showed a tendency for an increased total cell volume and a decreased numerical density; but, there was no conspicuous change in the total cell number. Determinations of the anti-bromodeoxyuridine (BrdU) and antiproliferative cell nuclear antigen (PCNA) antibody-positive cell counts showed a tendency for an increased proliferation rate for adrenomedullary chromaffin cells. Electron microscopy showed a slight increase in the number of Golgi apparatuses in these cells. Because the marked increase in urinary Ca excretion was concomitant with morphological changes that suggested the hyperfunction of chromaffin cells in the adrenal medulla and a tendency for an increased cell proliferation rate, we assume that persistent hyperfunction of the adrenomedullary chromaffin cells, which was mediated by enhanced Ca absorption from the intestinal tract, may have induced proliferative lesion. PMID- 7490789 TI - [Intravenous single dose toxicity of iodixanol, a new nonionic iso-osmolar contrast medium, in mice, rats and monkeys]. AB - The single-dose intravenous toxicities of iodixanol, a new nonionic iso-osmolar contrast medium, were investigated in mice, rats and monkeys. The LD50 values were estimated to be 17.9 gI/kg for male mice and 16.2 gI/kg for female mice, 18.8 gI/kg for male rats and 22.0 gI/kg for female rats, and more than 10.0 gI/kg for monkeys. There was no marked sex difference in mice or rats, nor any significant difference observed between these two rodent species. Decrease in spontaneous locomotor activity, ptosis, respiratory depression and abdominal posture were observed in many mice and rats. These signs disappeared mostly by 8 days after dosing in surviving animals. Death occurred between immediately and 4 days after dosing in mice, and between immediately and 14 days after dosing in rats. Transient depression of body weight gain was observed in the surviving mice and rats by 7 days after dosing. Histological examinations revealed congestion or hemorrhage in the renal medulla, vacuolation or necrosis of the renal proximal tubular epithelium in mice and rats that died and vacuolation of the renal proximal tubular epithelium in surviving rats. There were no significant treatment-related changes in the laboratory and pathological examinations in monkeys. PMID- 7490790 TI - [Toxicity of iodixanol, a new nonionic iso-osmolar contrast medium, to rats by repeated intravenous administration for 4 weeks with a 4-week recovery period]. AB - Groups of 10 male and 10 female Crl: CD (SD) BR rats were given iodixanol, a radiographic contrast medium, by intravenous injection at dosage levels of 30, 100 or 300 mgI/kg/day for four weeks to evaluate its toxicity. An extra 5 rats of each sex were given 300 mgI/kg/day for 4 weeks and then retained for 4 weeks without treatment to assess recovery. No clinical signs indicative of a toxic effect of iodixanol were noted. No effects of treatment with iodixanol on bodyweight gains, food consumption, laboratory investigations, ophthalmoscopic examinations, organ weights, bone marrow smears or macroscopic pathology were noted. Treatment related changes were noted microscopically in the kidneys only. Minimal cytoplasmic vacuolation in the proximal tubules was noted in a proportion of rats given 100 mgI/kg/day and all rats given 300 mgI/kg/day at the end of the treatment period. This effect was noted not to be fully reversible during four weeks off dose although recovery did occur in some rats. PMID- 7490791 TI - [Toxicity of iodixanol, a new nonionic iso-osmolar contrast medium, to cynomolgus monkeys by repeated intravenous administration for 4 weeks with a 4-week recovery period]. AB - Groups of 3 male and 3 female wild-caught cynomolgus monkeys were given iodixanol, a radiographic contrast medium, by intravenous injection at dosage levels of 100, 300 or 1,000 mgI/kg/day for four weeks to evaluate its toxicity. An extra 2 animals of each sex were given 1,000 mgI/kg/day for 4 weeks and then retained for 4 weeks without treatment to assess recovery. There were no deaths during the treatment period. Bruising at the injection sites was noted clinically and macroscopically was caused in part by the injection procedure itself and in part by the viscosity of the test formulation and the size of the dose. This finding is therefore, of no toxicological significance. No effects of treatment with iodixanol on body weights, food consumption, laboratory investigations, ophthalmoscopic examinations, organ weights or bone myelograms were noted. Cytoplasmic vacuolation of the kidney proximal tubules was noted with incidence and severity increasing with dosage level among animals given 100, 300 or 1,000 mgI/kg/day. This effect was noted not to be fully reversible at 1,000 mgI/kg/day after 4 weeks off-dose although reduced severity was noted and recovery was apparent in some animals. PMID- 7490792 TI - The susceptibility of five stains mice to infections with Angiostrongylus cantonensis. AB - The susceptibility to the infection of Angiostrongylus cantonensis was studied in five inbred stains of mice including four H-2 haplotypes. Three weeks after infection, C57 BL/10J (H-2b) and C57BL/6J (H-2b) stains of mice showed the lowest, CBA (H-2k) and A/J (H-2a) stains showed intermediate, and the BALB/cJ (H 2d) mice showed the greatest susceptibility to infection. BALB/cJ mice also displayed the most body weight loss, the highest number of worms and showed the least number of days of survival. The s susceptibility of C57BL/6J (H-2b) and C57BL/10J (H-2b) strains of mice sharing same H-2 haplotype was the same despite their different genetic backgrounds. These data suggested that the H-2 gene might influence the susceptibility of mice to A. cantonensis. PMID- 7490793 TI - In vitro production of interleukin-6 by human gingival, normal buccal mucosa, and oral submucous fibrosis fibroblasts treated with betel-nut alkaloids. AB - This study aimed to assess the possibility of a direct effect of betel-nut alkaloids arecoline and arecaidine on cell proliferation and interleukin-6 (IL-6) production by cultured fibroblasts from human normal gingiva, buccal mucosa and oral submucous fibrosis (OSF) buccal mucosa in vitro. Confluent monolayers of fibroblasts were incubated with or without alkaloids in the presence of 10% fetal calf serum for 48 h at 37 degree C in 5% CO2 and air. At the end of the culture period, supernatants were collected and assayed for IL-6 level. The cell proliferation was monitored by determining 5-bromo-2'-deoxy-uridine (BrdU) incorporated into cellular DNA. Except for the fact that arecoline inhibited cell growth at 100 micrograms/ml, arecoline and arecaidine had similar dose-dependent stimulant effects on the proliferation of these three groups fibroblasts. Concentrations of IL-6 in the control culture supernatants were greatest in healthy gingival fibroblasts, followed by normal buccal mucosa and OSF. Also, the presence of fetal calf serum could stimulate IL-6 release. Except for arecoline at the 100 microgram/mg, there were no significant differences in IL-6 levels between treated and control cultures of the same group when the data were expressed with mean +/- S.D.. However, two of six individuals' normal buccal mucosa fibroblasts significantly released less IL-6, and some cases of OSF and healthy gingiva exhibited slightly higher levels of IL-6 when cells were exposed to arecoline or arecaidine in cultures. Such findings suggests that arecoline and arecaidine can enhance cell proliferation and affect fibroblasts to synthesize IL 6. Furthermore, IL-6 may be a contributing molecular factor in the pathological features noted in OSF. PMID- 7490794 TI - Utility of Argyrophilic nucleolar organizer regions (AgNOR) in differentiating malignant from benign of colorectal tumors. AB - BACKGROUND: Argyrophilic nucleolar organizer regions (AgNOR) can reflect the activity of cellular proliferation. The purpose of this study is to evaluate the potential value of AgNOR in differentiating benign from malignant colon epithelial neoplasms, and to determine the correlation between the nucleus AgNOR and the grade of colonic adenocarcinomas. METHODS: In this study, AgNOR technique was applied to 61 paraffin embedded sections of colorectal tissue including normal mucosa (n = 10), adenomatous polyp (n = 16), and adenocarcinoma (n = 35). RESULTS: The mean +/- standard error (SE) numbers of AgNOR dots per nucleus of normal mucosa, adenomatous polyp, and adenocarcinoma were 2.17 +/- 0.07 (n = 10) 3.89 +/- 0.10 (n = 16) and 5.52 +/- 0.10 (n = 35), respectively (p < 0.00001). In addition, the mean numbers of AgNOR dots per nucleus of well differentiated (WD) adenocarcinoma (n = 14), moderately differentiated (MD) adenocarcinoma (n = 11) and poorly differentiated (PD) adenocarcinoma (n = 10) were 5.20 +/- 0.12, 5.81 +/- 0.20, 5.67 +/- 0.15, respectively. MD and PD tumor had significantly higher AgNOR count than that of WD tumor (p < 0.05). However, there was no significant difference between MD and PD colorectal adenocarcinoma. CONCLUSION: AgNOR method is a simple, rapid method in diagnosis of colorectal tumors, and it provides a useful adjunct to histopathology in the diagnosis of colorectal tumors. PMID- 7490796 TI - Postoperative diaphragmatic dysfunction in patients undergoing open-heart surgery. AB - Diaphragmatic elevation after cardiac surgery may result in compromized post operative respiratory function, prolonged ventilator use, longer intensive care unit (ICU) stays, poor quality of life and even mortality. 200 patients receiving open-heart surgery during the two years from October 1990 to October 1992 were studied retrospectively. 26 patients developed postoperative diaphragmatic elevation (Group 2). On the other hand, 174 patients did not (Group 1). The mean age of Group 1 (37.3 +/- 20.0 years) was younger than that of Group 2 (57.4 +/- 9.1 years), P < 0.0001. The mean aortic cross-clamp time was 70.1 +/- 38.1 minutes in Group 1. On the other hand, Group 2 had a longer crossclamp time (84.5 +/- 31.3 Minutes), p < 0.03. Our study revealed that the patients who suffered post operative diaphragmatic dysfunction were older and had a longer aortic crossclamp time than the patients who did not. PMID- 7490797 TI - Yolk sac tumor of unknown origin with intracranial and spinal metastases. AB - Primary intracranial yolk sac tumor (Endodermal sinus tumor, YST) is quite rare, and most usually involves the pineal gland. This report concerns a rare case of unknown origin of yolk sac tumor with intracranial and spinal metastases. The 6 year-old boy initially manifested symptoms of acute urinary retension and paraparesis. Computed tomography (CT) and magnetic resonance imaging (MRI) revealed an intrathecal tumor spreading below the third lumbar segment, and tumors also appeared in the left temporal area and right cerebellar hemisphere. The pineal region was free of neoplastic involvement and the gonads were normal. The patients underwent operation to remove intracranial and spinal tumors, and postoperative radiotherapy was administered. The histological findings showed metastatic yolk sac tumor. Multiple intracranial and spinal seeding were noticed postoperatively. The patient died of recurrent YST and pneumonia three months after operation. The origin of secondary YST can always be found in specific midline sites. This is a rare case since the primary lesion is unknown. PMID- 7490795 TI - Frozen section of diagnosis of breast lesions. AB - Frozen section diagnosis rendered in 549 consecutive breast biopsies performed in 5 years in a single pathology laboratory was correlated with the final pathological diagnosis. There were no false positive reports among the 220 (40.1%) biopsies interpreted as benign lesions in paraffin sections. Among 329 (59.9%) malignant biopsies on paraffin sections, 3 cases were interpreted as benign lesions on frozen sections. Three false negatives included 2 ductal carcinoma in situ and one infiltrating ductal carcinoma associated with papillomatosis. The tumors were small and confined to the breast without any evidence of metastasis. There was a very good correspondence between the frozen section diagnosis and the paraffin section diagnosis (K = 0.98). The sensitivity of frozen section diagnosis was 99.1% and the clinical diagnostic specificity was 100%. Our results suggest that frozen section diagnosis is a highly reliable procedure, but small lesions (less than 1 cm in diameter, or non-palpable) should not be subjected to frozen section examination to avoid unnecessary loss of neoplastic tissue during the frozen section. The careful investigation of paraffin-embedded tissue is recommended for small breast lesions in breast conserving lumpectomy. PMID- 7490798 TI - [Incidence of non-arteritic anterior ischemic optic neuropathy in glaucomatous patients]. AB - Raised intraocualr pressure has been suggested to be a contributing factor in NAION occurrence. Thus, there should be a high NAION incidence rate in glaucoma patients when compared with that of the general population. From June 1990 to December 1991, we retrospectively surveyed 180 glaucoma patients to find the incidence of NAION, but no NAION cases were found. We concluded that the increase in intraocular pressure is not a major factor to cause NAION. PMID- 7490799 TI - [Ulcerative colitis in Taichung Veterans General Hospital: a clinical study]. AB - This report concerns 34 patients of ulcerative colitis admitted to Taichung Veterans General Hospital, from 1983 to 1994. Among them 26 were male and 8 were female. The age at onset were mostly from 50 to 60. The average duration between onset of symptoms and the date of definite diagnosis was 10 months. The most common presenting symptom was bloody diarrhea (64.7%). Most of our patients were in the moderately severe group of disease (67.6%), according to the severity defined by Truelove and Witts. The most frequent endoscopic findings of mucosa was classified as Grade III (38.2). Descending colon (91.2), rectum (85.3%), and sigmoid colon (88.2%) were the most frequently involved areas. The major clinical course were chronic intermittent and chronic continuous type (55.9%). Extraintestinal manifestations were found in 2 cases: one was found in the skin, and the other in the joint, respectively. Treatment of ulcerative colitis in our series was mainly medical (91.2%). However, 3 patients received emergent surgical intervention, and 10 patients finally underwent operation because of major complications or failure to respond to medical treatment. PMID- 7490800 TI - Collagen injection for intrinsic sphincteric deficiency in men. AB - PURPOSE: We evaluated the efficacy of collagen injections in the treatment of male urinary incontinence due to intrinsic sphincteric deficiency. MATERIALS AND METHODS: A total of 88 men (mean age 68 years) with mild to severe intrinsic sphincter deficiency underwent a mean of 3.5 transurethral injections of collagen (mean total volume injected 25 ml.). Patients were subdivided into 2 groups based on use of more than 3 or 3 or fewer pads per day. Patient age, pad use before treatment, duration of leakage, number of injections, volume of collagen used and etiology of incontinence were compared for the 2 groups. RESULTS: Of the patients 42 became nearly completely dry, 19 had substantial improvement but still required 1 to 3 pads per day, 14 consistently used fewer pads but still more than 3 per day and 13 showed no improvement. Most injections were performed with the patient under local anesthesia and no significant morbidity occurred. CONCLUSIONS: In select patients collagen injections appear to be effective, easy to deliver and safe. Pretreatment incontinence severity, concomitant detrusor abnormalities and etiology of intrinsic sphincteric deficiency other than radical retropubic prostatectomy were associated with a worse response to collagen therapy. PMID- 7490801 TI - Early complications and survival following short-term palliative radiotherapy in invasive bladder carcinoma. AB - PURPOSE: We studied the early side effects and survival rates after short-term radiotherapy (7 Gy. 3 times or 5 Gy. 4 times) in patients with muscle invasive bladder carcinoma. MATERIALS AND METHODS: Between 1981 and 1992, 96 patients (median age 80 years) were treated. Followup was complete until 1994 or death. RESULTS: Early side effects caused the hospitalization of 22 patients and may have contributed to the death of 5 elderly patients. Median survival for patients with stages T2M0, T3M0, T4M0 and T2 to 4M+ disease was 27 months, 6.3 months, 5.6 months and 2.9 months, respectively. CONCLUSIONS: Short-term radiotherapy is of doubtful benefit to elderly patients with advanced bladder carcinoma and may even be harmful. PMID- 7490802 TI - Bladder carcinoma--the good, the bad and the ugly. PMID- 7490803 TI - Detection of early bladder cancer by 5-aminolevulinic acid induced porphyrin fluorescence. AB - PURPOSE: We determined whether the sensitivity of detecting dysplasia or early bladder cancer can be improved by 5-aminolevulinic acid induced porphyrin fluorescence. MATERIALS AND METHODS: A 3% 5-aminolevulinic acid solution was instilled intravesically before cystoscopy in 104 patients. The 5-aminolevulinic acid induced porphyrin fluorescence was excited by violet light from a krypton ion laser (wavelength 406.7 nm.). RESULTS: The sensitivity of the fluorescence cystoscopy (96.9%) was significantly (p < 0.0001) greater than that of white light cystoscopy (72.7%). There was no impact on specificity. CONCLUSIONS: Due to the high sensitivity of the procedure fluorescence guided biopsies are recommended instead of random biopsies. PMID- 7490804 TI - Pattern of metastases in relation to characteristics of primary tumor and treatment in patients with disseminated urothelial carcinoma. AB - PURPOSE: The anatomical pattern of recurrence and metastases in patients with urothelial cancer are described, and the relationship between treatment and features of the primary invasive tumor and the subsequent pattern of metastases is analyzed. MATERIALS AND METHODS: Between 1976 and 1991, 240 patients with recurrent or metastatic urothelial cancer were admitted to our department. RESULTS: The majority of the patients had recurrence within 2 years after initial diagnosis. Local recurrences and lung metastases were diagnosed significantly earlier than other metastases. Multiple sites were involved in more than half of the patients. The most common sites of recurrence were local in the bladder in 65% of the cases and bone in 35%, followed by lymph nodes in 26% and lung in 20%. The pattern of metastases was similar in patients with different histological findings, grade and location of the primary tumor. Patients younger than 60 years and those with cancer of the renal pelvis more often had distant metastases compared to older patients with bladder cancer. Local recurrences were less frequent in patients who had undergone cystectomy compared to those treated with radiotherapy only. Moreover, patients with local recurrences were likely to have metastases elsewhere. CONCLUSIONS: Bone was the most frequent site of metastases outside the pelvis and all patients suspected to have recurrence should be examined for bone metastases. The results indicate that the pattern of recurrence and metastases are not dependent on the features of the primary tumor. PMID- 7490805 TI - Postoperative radiation therapy in 26 patients with invasive transitional cell carcinoma of the upper urinary tract: no impact on survival? AB - PURPOSE: To evaluate the role of adjuvant radiation therapy in invasive transitional cell carcinoma of the upper urinary tract, we retrospectively reviewed a series of 26 patients who underwent radical surgery plus postoperative prophylactic irradiation for such a tumor. MATERIALS AND METHODS: Between February 1980 and October 1993, 18 men and 8 women (mean age 65 +/- 9 years, standard deviation) were treated for an invasive transitional cell carcinoma of the upper urinary tract. Tumor location was the renal pelvis in 15 patients (58%). The tumor was pathological stage B in 11 patients (42%) and stage C in 15 (58%). Tumor grade was 2 in 10 patients, 3 in 15 and unknown in 1. One patient had epidermoid metaplasia of urothelial cancer and 9 had node involvement. All patients underwent surgery followed by radiation therapy to a total dose of 45 Gy. to the tumor bed (23) and/or regional nodes (18). RESULTS: After a mean followup of 45 months 13 patients (50%) were alive and 11 were disease-free at analysis. Local tumor relapse, nodal recurrence and metastasis were noted in 1, 4 (15%) and 14 (54%) patients, respectively. All patients with nodal recurrence had metastasis. A secondary location was noted frequently (6 bladder, 1 contralateral renal pelvis and 1 urethral tumors). Overall 5-year survival rate and 5-year survival rate with no evidence of disease were 49% and 30%, respectively. Overall 5-year survival rates were 60% for stage B and 19% for stage C disease (p = 0.07), 49% for node-negative versus 15% for node-positive cancer (p = 0.04), and 90% for grade 2 and 0% for grade 3 tumors (p < 0.01). CONCLUSIONS: In our trial using a radio-surgical approach, local control of disease and survival rates were similar to those reported previously in surgical series. Prophylactic postoperative radiation therapy is not recommended except in prospective randomized studies. PMID- 7490806 TI - Adjuvant methotrexate, vinblastine and cisplatin chemotherapy for invasive transitional cell carcinoma: Taiwan experience. AB - PURPOSE: The feasibility of adjuvant cisplatin, methotrexate and vinblastine chemotherapy was evaluated in Taiwanese patients with invasive transitional cell carcinoma at high risk for recurrence. MATERIALS AND METHODS: We assigned 56 patients with high risk transitional cell carcinoma (vascular or lymphatic invasion in the primary tumor, poorly differentiated stage P2, P3, P4 or N+ and M0) to receive adjuvant chemotherapy after radical urological surgery. The chemotherapy consisted of 40 mg./m.2 methotrexate and 4 mg./m.2 vinblastine on days 1 and 8, and 100 mg./m.2 cisplatin on day 2 given in 6 courses at 21-day intervals. RESULTS: Median followup was 44 months. An average of 4.63 cycles of chemotherapy was administered. The median actual survival was 44 months, and the 1 and 3-year survival probabilities were 92% and 50%, respectively. The median disease-free survival was 15.5 months, and the 1 and 3-year disease-free survival probabilities were 66% and 28%, respectively. Only 5 (9%) and 1 (2%) patients had grades 3 and 4 leukopenia, respectively, and none died of sepsis. CONCLUSIONS: The use of adjuvant cisplatin, methotrexate and vinblastine chemotherapy in patients with invasive transitional cell carcinoma at high risk for recurrence is feasible with tolerable toxicity but randomized controlled trials will be required to assess the benefit. PMID- 7490807 TI - Urothelial malignancy--much promise but little progress. PMID- 7490808 TI - Dorsal free graft urethroplasty. AB - PURPOSE: Dorsal free graft urethroplasty was performed to reduce the incidence of urethrocele. MATERIALS AND METHODS: We treated 12 patients with penile and 13 with bulbous strictures. Of the 13 patients with a bulbous stricture 6 received a dorsally placed tube graft and 7 received a patch graft. RESULTS: Temporary fistulas were seen on postoperative urethrography in 5 cases but they all resolved spontaneously. At a mean followup of 35.8 months clinical and radiological findings were excellent in 23 cases and good in 2. No signs of graft weakening, such as post-void dribbling or diminished ejaculation, were apparent. CONCLUSIONS: The use of free skin grafts for urethral reconstruction is anatomically healthier in the dorsal than in the ventral position. PMID- 7490809 TI - Intravaginal stimulation randomized trial. AB - PURPOSE: The effectiveness of intravaginal electrical stimulation was compared to standard therapy in the treatment of genuine stress urinary incontinence and detrusor instability. MATERIALS AND METHODS: A total of 57 women with urinary incontinence was evaluated with video urodynamics and voiding diaries before and after treatment. Of the women 18 with stress urinary incontinence were randomized to electrical stimulation or Kegel exercise and 38 with detrusor instability were randomized to anticholinergic therapy or electrical stimulation. RESULTS: Of patients using electrical stimulation in the stress urinary incontinence group 66% improved and 72% of the patients with detrusor instability treated with electrical stimulation improved. These rates were not statistically significant when compared to traditional therapy. CONCLUSIONS: Electrical stimulation is safe and at least as effective as properly performed Kegel and anticholinergic therapy in the treatment of stress urinary incontinence and detrusor instability. PMID- 7490810 TI - Split-thickness skin graft urethroplasty and tunica vaginalis flaps for failed hypospadias repairs. AB - PURPOSE: Via a 2-stage procedure, 10 patients with failed hypospadias repairs were treated by a varied combination of split-thickness mesh graft urethroplasty and tunica vaginalis flap. MATERIALS AND METHODS: A bed for the mesh graft in 3 patients was provided by a tunica vaginalis flap. Tunica vaginalis flaps were also used as an intermediate layer during stage 2 of the repair. RESULTS: No strictures or fistulas occurred in 8 patients. Two patients await stage 2 repair after successful stage 1 placement of the mesh graft. CONCLUSIONS: The combination of split-thickness mesh graft urethroplasty and a tunica vaginalis flap appears to achieve success in the difficult patient with complex hypospadias subsequent to multiple failed repairs. PMID- 7490811 TI - Ultrex cylinders: problems with uncontrolled lengthening (the S-shaped deformity). AB - PURPOSE: We investigated whether the AMS Ultrex cylinder, developed to provide cylinder lengthening while controlling girth expansion, acted as a tissue expander and, thus, promoted penile lengthening. MATERIALS AND METHODS: Between December 1990 and January 1992, 59 AMS Ultrex prostheses were implanted. Cylinder length corresponded to intracorporeal measurements. The results of lengthening and occurrence of an S-shaped deformity were statistically analyzed. RESULTS: No penile lengthening occurred. In 19 patients (32%) cylinder replacement was required for the S-shaped deformity. CONCLUSIONS: Ultrex cylinders should be downsized and prolonged inflation should be avoided. They should not be implanted in cases of Peyronie's disease, or to replace malleable or self-contained inflatable implants. PMID- 7490812 TI - Penile scarring with intracavernous injection therapy using prostaglandin E1: a risk factor analysis. AB - PURPOSE: We attempted to identify risk factors for penile scarring in patients using prostaglandin E1 injection therapy. MATERIALS AND METHODS: Records of 92 patients using prostaglandin E1 therapeutically were reviewed. With statistical methods, patients with (15) and without (77) scarring were compared. RESULTS: No significant differences between the 2 groups were found regarding duration of followup, injection frequency, prostaglandin E1 dose per injection, total number of injections or total prostaglandin E1 dose. Patients with initial penile scarring did not have a significantly higher incidence of further scarring with prostaglandin E1. CONCLUSIONS: Penile scarring with prostaglandin E1 injection therapy is sporadic and unpredictable. Patients with initial penile scarring do not have a higher incidence of further scarring. PMID- 7490813 TI - Dihydroxyphenylglycol in pheochromocytoma: its diagnostic use for norepinephrine dominant tumor. AB - PURPOSE: To discern whether the 3,4-dihydroxyphenylglycol produced in adrenal medulla is associated with altered urinary excretion, we compared the tissue and urinary levels of this catecholamine metabolite, epinephrine and norepinephrine in 23 patients with pheochromocytoma and 26 subjects with renal cell carcinoma. MATERIALS AND METHODS: Tissue and urine concentrations of dihydroxyphenylglycol, epinephrine and norepinephrine were determined by catechol-O-methyl-transferase based radioenzymatic method and high performance liquid chromatography with electrochemical detection. RESULTS: Contents of dihydroxyphenylglycol, epinephrine and norepinephrine in pheochromocytomas were higher than those in normal adrenal medullae. Removal of pheochromocytomas lowered urinary excretion of these catecholamines and dihydroxyphenylglycol. Norepinephrine content correlated closely with dihydroxyphenylglycol content in normal adrenal medullae and pheochromocytomas but not with epinephrine content in either tissue. Contents of norepinephrine and dihydroxyphenylglycol correlated significantly with urinary norepinephrine excretion and dihydroxyphenylglycol excretion in patients with pheochromocytoma but not in subjects with renal cell carcinoma. The norepinephrine-to-dihydroxyphenylglycol ratio in a 24-hour preoperative urine collection from patients with pheochromocytoma was higher than that from subjects with renal cell carcinoma, which was almost similar to that of epinephrine dominant pheochromocytomas. CONCLUSIONS: Our study suggests that dihydroxyphenylglycol in urine in subjects with renal cell carcinoma is predominantly of a neuronal origin and urinary dihydroxyphenylglycol in patients with pheochromocytoma is of tumor origin. A high level of norepinephrine-to dihydroxyphenylglycol ratio in urine can be used to determine the diagnosis of the norepinephrine dominant type pheochromocytoma. Measurement of at least a few markers is preferable for precise biochemical diagnosis of pheochromocytoma. PMID- 7490815 TI - A randomized crossover study using yohimbine and isoxsuprine versus pentoxifylline in the management of vasculogenic impotence. AB - PURPOSE: We evaluated the efficacy and safety of yohimbine and isoxsuprine or pentoxifylline in the management of vasculogenic erectile dysfunction. MATERIALS AND METHODS: A total of 20 patients diagnosed with arterial insufficiency and cavernous venous leakage by pharmacological penile duplex scanning underwent nonhormonal oral therapy. A randomized crossover study was performed using 5.4 mg. yohimbine plus 10 mg. isoxsuprine or 400 mg. pentoxifylline 3 times daily. Response to therapy was assessed by a sexual questionnaire and repeat penile duplex ultrasonography. RESULTS: No patient in either phase of the study in either group had a complete response to the oral regimen(s) and there was no improvement in the pre-injection or post-injection cavernous arterial peak systolic flow velocities or resistance indexes. CONCLUSIONS: Our study suggests that these oral agents, although well tolerated, were not effective in the management of these patients with mixed vasculogenic erectile dysfunction. PMID- 7490816 TI - Impotence--defining the role of minimally invasive therapy. PMID- 7490814 TI - Intracorporeal phenylephrine in the treatment of priapism. AB - PURPOSE: The effectiveness and complications of intracorporeal phenylephrine without aspiration or irrigation as a treatment for priapism were assessed. MATERIALS AND METHODS: Nine consecutive patients who presented with priapism were treated with 0.5 mg. phenylephrine diluted in 2 cc normal saline injected directly into the corpus cavernosum. Blood pressure and pulse were measured before and after injection, and monitored every 15 minutes. If no detumescence was noted after 15 minutes the injection was repeated. Neither penile blood aspiration nor irrigation was performed. RESULTS: Of 9 patients 8 responded with detumescence. The number of injections required ranged from 1 to 6. No changes in blood pressure, heart rate or side effects were observed. CONCLUSIONS: We believe that this is a safe and effective method for treatment of priapism. Routine initial aspiration or irrigation of the corpora before intracorporeal injection of alpha-adrenergic agents does not appear to be necessary. PMID- 7490817 TI - Penile fractures: evaluation, therapeutic approaches and long-term results. AB - PURPOSE: Traumatic rupture of the corpus cavernosum of the penis is rare and has been reported infrequently. We demonstrated the moderate frequency and most common etiologies of this condition. MATERIALS AND METHODS: Between February 1990 and June 1993, 68 patients underwent an emergency operation 3 hours to 4 days after penile fracture. RESULTS: Among 32 patients who completed a questionnaire functional results were excellent except for 3 with penile curvature and pain during coitus who underwent surgery more than 48 hours after injury. CONCLUSIONS: Fracture of the penis is a common urological problem. In our series the most common cause was manipulation. To avoid serious complications immediate surgical intervention is recommended. PMID- 7490818 TI - Could standardized cavernosometry be helpful in therapeutic management of veno occlusive dysfunction? AB - PURPOSE: Due to the lack of a gold standard for performing cavernosometry, we selected 30 patients with veno-occlusive dysfunction to evaluate the internal relationships of cavernosometric parameters and their reliability for therapeutic decisions. MATERIALS AND METHODS: Cavernosometry was performed after injection of 20 to 40 micrograms of prostaglandin E1. Maintenance flow rates, intracavernous pressure decay following cessation of flow and intracavernous pressure changes after compression maneuvers were the main parameters considered. RESULTS: Cavernosometric results were standardized as grade 1-7 patients with maintenance flow rate less than 20 ml. per minute (mean 18 +/- 2.5) and intracavernous pressure decay 27.5 +/- 15%, grade 2-11 with maintenance flow rates significantly lower (p < 0.001) than grade 1 (mean 37 +/- 11 ml. per minute) and intracavernous pressure decay 33 +/- 20%, and grade 3-12 with only recorded induction flows greater than 70 ml. per minute and intracavernous pressure less than 50 mm. Hg. A strong statistical correlation (p < 0.01) was noted between maintenance flow rate and percent of intracavernous pressure decays. CONCLUSIONS: These parameters allowed us to choose different therapies, such as complex venous surgery, mixed pharmacotherapy or prosthetic implants. Good sexual function was restored in 85% of the cases. We can conclude that our model of standardized cavernosometry has made easier the therapeutic choices in patients who do not respond to intracavernous drugs. PMID- 7490820 TI - Erections--broken, leaking and salvaged. PMID- 7490819 TI - The prosthesis salvage operation: immediate replacement of the infected penile prosthesis. AB - PURPOSE: We describe our experience with salvage of the infected penile prosthesis at initial presentation in 11 patients. MATERIALS AND METHODS: All patients with prosthesis infection who presented since 1991 were considered for salvage surgery. Contraindications to a salvage operation included necrotic infections, diabetic patients with purulence in the corporeal bodies, rapidly developing infections and erosion of the device cylinders. RESULTS: In 1 patient in this group a salvage attempt was repeated after re-infection, for an overall success rate of 91%. Mean followup for the group was 21 months (range 9 to 42). Staphylococcus epidermidis was the infecting organism in 75% of our patients. CONCLUSIONS: Our experience demonstrates the safety and advantages of the immediate salvage technique. PMID- 7490821 TI - The immunological features of Peyronie's disease. AB - PURPOSE: We investigated the immunological features and possible autoimmune basis of Peyronie's disease. MATERIALS AND METHODS: The sera of 100 patients with Peyronie's disease were tested for circulating autoantibodies, including anti penis antibodies, by indirect immunofluorescence. Antibody deposition and the immunological activity in Peyronie's plaque tissue from patients with early and long-standing disease were also assessed. RESULTS: Circulating anti-penis antibodies were not found in any patient although antinuclear antibodies were present in 24%. Patients with early Peyronie's disease had IgM antibody deposition, marked T lymphocytic and macrocytic infiltration in the sub-tunical space, increased expression of adhesion molecules by endothelial cells and an increased human lymphocyte antigen class 2 expression by the cellular infiltrate, indicating cellular immune activation. CONCLUSIONS: These results show that some of the features of autoimmunity, in particular the cell mediated response, are present in Peyronie's disease. PMID- 7490822 TI - The effect of cocaine and its metabolites on Sertoli cell function. AB - PURPOSE: We determined the in vitro effects of cocaine and its 2 major metabolites, benzoylecgonine and ecgonine, on Sertoli cell function. MATERIALS AND METHODS: Sertoli cells were isolated from 18 to 20-day-old Sprague-Dawley rats. Sertoli cells were incubated with 4 concentrations (6.25 x 10(-5), 2.5 x 10(-4), 1 x 10(-3) and 4 x 10(-3) M./l.) of cocaine, benzoylecgonine and ecgonine for 24, 48 and 72 hours. Transferrin and androgen-binding protein secretion was measured to determine the effect of cocaine and its metabolites on Sertoli cell function. RESULTS: Cocaine, benzoylecgonine and ecgonine had the ability to decrease transferrin secretion. The 2 highest doses of cocaine (1 x 10(-3) and 4 x 10(-3) M./l.) significantly decreased transferrin production at 24, 48 and 72 hours. The effects of benzoylecgonine and ecgonine on transferrin production were less pronounced than those of cocaine. Androgen-binding protein secretion was also decreased by exposure to cocaine and its metabolites although the decrease was less marked compared to that of transferrin production. CONCLUSIONS: We demonstrated in vitro harmful effects of cocaine and its 2 major metabolites on Sertoli cell function, as measured by transferrin and androgen-binding protein production. PMID- 7490823 TI - Presence of sperm in the pre-vasectomy reversal semen analysis: incidence and implications. AB - PURPOSE: We determined the incidence of sperm in the ejaculate before vasectomy reversal, and correlated this result with intraoperative and postoperative findings. MATERIALS AND METHODS: Before vasectomy reversal semen was analyzed and granulomas were palpated in 186 men. The results were correlated with intraoperative vasal fluid and postoperative semen analyses. RESULTS: Of 18 men (9.7%) with sperm present in the pre-reversal analysis 94% had sperm in at least 1 vas intraoperatively. The presence of palpable granulomas at the vasectomy site did not correlate with either pre-reversal or post-reversal semen analyses, or the presence of sperm in the vasal fluid intraoperatively. CONCLUSIONS: Sperm are present in 9.7% of pre-reversal ejaculates and predict sperm in at least 1 vas intraoperatively. These findings also suggest the possibility of late vasectomy failures. PMID- 7490824 TI - The secondary internal ring: applications to surgery of the inguinal canal. AB - PURPOSE: In 1975 the clinical importance of the preperitoneal fascial or secondary internal ring was noted based on procedures performed through the suprainguinal preperitoneal approach. The importance of knowledge of the secondary internal ring in the performance of inguinal hernia repairs, orchiopexies and operations for impalpable testes through the inguinal canal is shown, along with a description of the anatomy of the internal ring and subjacent retroperitoneal connective tissue. MATERIALS AND METHODS: More than 250 consecutive inguinal canal dissections were performed during surgery for inguinal hernia repair, orchiopexy or impalpable testes by a single surgeon using 3.5x magnification. RESULTS: The intraoperative dissections clearly showed the existence of a secondary internal ring, which when opened provided wide access to the retroperitoneal space containing the internal spermatic vessels and vas deferens. CONCLUSIONS: Knowledge of the anatomy of the secondary internal ring is an adjunct to the performance of operations through the inguinal canal. PMID- 7490825 TI - Ultrasonographic characteristics of testicular adenomatoid tumors. AB - PURPOSE: We determined the characteristics of testicular adenomatoid tumors which could distinguish these lesions from testicular malignancies. MATERIALS AND METHODS: Clinical presentation and ultrasonographic findings in 3 men with testicular adenomatoid tumors were compared with those in 10 men treated for testicular malignancies. RESULTS: Clinical presentation was similar for all patients. Of the 3 adenomatoid tumors 2 appeared isoechoic on ultrasound and 1 appeared normal. None of the 10 cancers appeared isoechoic or normal on ultrasound. CONCLUSIONS: Small tumors that appear isoechoic on ultrasound should be biopsied through an inguinal incision with frozen section assessment rather than immediate radical orchiectomy. PMID- 7490826 TI - Decrease in mortality from benign prostatic hyperplasia: a major unheralded health triumph. AB - PURPOSE: A systematic examination of all available mortality data from benign prostatic hyperplasia (BPH) between 1950 and 1990 was done to estimate the changing international mortality pattern of this condition. MATERIALS AND METHODS: Mortality data in which BPH was the underlying cause of death were abstracted from the World Health Organization mortality data base. These data were available from a variety of international countries (in many since 1950). RESULTS: Mortality rates have decreased with considerable magnitude in developed western countries between the early 1950s and late 1980s. This fact could reasonably be attributed to the improved management of the most severe complications of BPH and to improvements in surgery and anesthesia, which have made surgical interventions of the prostate possible in a greater proportion of men, and safer in the immediate and subsequent postoperative period. CONCLUSIONS: The decreases noted in western countries, such as the United Kingdom (where 8,700 fewer men die each year presently than would be expected if the BPH mortality rates from the early 1950s still applied), United States (13,681 fewer deaths) and France (2,884 fewer deaths), indicate a considerable but unheralded achievement for modern medicine. Unfortunately, these decreases have not been observed to the same extent in central and eastern Europe and South America, where the residual high mortality rates could be lowered by education, and the widespread availability of modern surgical and anesthetic equipment. PMID- 7490827 TI - Neodymium:YAG laser coagulation prostatectomy: 3 years of experience with 227 patients. AB - PURPOSE: For 3 years we prospectively followed a cumulative cohort of men who underwent neodymium:YAG laser coagulation prostatectomy to relieve bladder outlet obstruction to determine the safety, efficacy and durability of this procedure. MATERIALS AND METHODS: A total of 227 men with symptomatic bladder outlet obstruction was treated with the Urolase side-firing laser fiber between October 15, 1991 and October 15, 1994. Voiding outcomes, including peak urinary flow rates, residual urine volumes and American Urological Association symptom scores, were measured with time, and immediate and long-term complications were assessed. RESULTS: Median patient followup was 26 months (maximum 42). Significant improvement occurred in all measured voiding parameters, including 133% in peak flow rates and 61% in symptom scores 1 year postoperatively. Overall 87% of patients noticed improved quality of life as a result of surgery. Minimum postoperative followup was 6 months in all cases. Complications included prostatitis in 2.6% of patients, urethral stricture in 1.8%, bladder neck contracture in 4.4% and reoperation for residual prostate tissue in 5.3%. CONCLUSIONS: Neodymium:YAG laser coagulation of the prostate represents an efficacious surgical intervention for symptomatic bladder outlet obstruction with minimal associated morbidity. Voiding outcomes are durable through 3 years. PMID- 7490828 TI - The outcome of renal ultrasound in the assessment of 556 consecutive patients with benign prostatic hyperplasia. AB - PURPOSE: We determined the additional value of renal ultrasonography in the assessment of patients with benign prostatic hyperplasia. MATERIALS AND METHODS: Renal ultrasound was performed in 556 consecutive patients with benign prostatic hyperplasia and the results were correlated with other clinical parameters. RESULTS: Of the patients 14 (2.5%) had dilatation of the renal pelvis, 65 (11.7%) had renal cysts and 1 (0.18%) had renal cell carcinoma. The serum creatinine level appeared to be correlated with dilatation of the renal pelvis. To predict dilatation additional information can be obtained by including the results of the post-void residual measurements. CONCLUSIONS: Renal ultrasound is indicated only in patients with a specific serum creatinine level and/or post-void residual volume. PMID- 7490830 TI - Benign prostatic hyperplasia: practice makes perfect. PMID- 7490829 TI - Interleukin-2 based home therapy of metastatic renal cell carcinoma: risks and benefits in 215 consecutive single institution patients. AB - PURPOSE: In 215 consecutive patients with advanced metastatic renal cell carcinoma seen at a single institution the efficacy and tolerance of different subcutaneous recombinant interleukin-2 based home therapies were assessed. MATERIALS AND METHODS: Treatment consisted of subcutaneous recombinant interleukin-2 alone and subcutaneous recombinant interleukin-2 in combination with recombinant interferon-alpha 2 with or without intravenous 5-fluorouracil. RESULTS: Overall objective response rate in 215 patients was 33% (95% confidence interval 26 to 39%). Among 16 patients receiving recombinant interleukin-2 alone there was 1 partial remission (overall response 6%). In 79 patients receiving recombinant interleukin-2 and interferon-alpha 2 in combination 6 complete and 16 partial remissions occurred (overall response 28%). Of 120 patients receiving a combination of recombinant interleukin-2, recombinant interferon-alpha 2 and 5 fluorouracil 13 achieved a complete and 34 a partial remission (overall response 39%). Of all patients 5% achieved long-lasting remissions and remain disease free. Multivariate analyses identified pretreatment erythrocyte sedimentation rate greater than 70 mm. per hour and lactic dehydrogenase greater than 280 units per l. as independent prognostic factors of major significance (p < or = 0.0001) in metastatic renal cell carcinoma. Additionally, neutrophil count greater than 6,000/microliters., hemoglobin less than 100 gm./l., extrapulmonary metastases and bone lesions were identified as minor (p < or = 0.006) prognostic variables. Patients were assigned to 1 of 3 risk categories according to cumulative risk score defined as the function of the sum of all 6 independent variables. In 116 intermediate risk patients 2-year survival was 65% (median survival not reached after 32 months) with recombinant interleukin-2, recombinant interferon-alpha 2 and 5-fluorouracil, as opposed to 27% 2-year survival (median survival 15 months) with recombinant interleukin-2 and interferon-alpha 2 (p < 0.0001), and 0% (median survival 4.8 months) with single agent recombinant interleukin-2. In the majority of patients systemic toxicity of subcutaneous recombinant interleukin-2 based protocols was limited to grade 1 or 2 constitutional symptoms, that is fever, chills, malaise and anorexia, which allowed for outpatient therapy. CONCLUSIONS: The present outpatient recombinant interleukin-2 triple drug combination protocol was as effective as the most aggressive intravenous recombinant interleukin-2 regimen available. Combination home therapy eliminated the need for inpatient and/or intensive care as required for intravenous cytokine administration and, thereby, it substantially improved the therapeutic index and cost-effectiveness of recombinant interleukin-2 therapy in metastatic renal cell carcinoma stratified for risk. PMID- 7490831 TI - Factors affecting change in quality of life after prostatectomy for benign prostatic hypertrophy: the impact of surgical techniques. AB - PURPOSE: The impact of prostatectomy on quality of life was assessed in patients with benign prostatic hypertrophy (BPH) who were classified according to the expected benefit from surgical intervention. The relative impact of the 2 surgical techniques (open versus closed) on short-term quality of life was compared. MATERIALS AND METHODS: An observational study was done on 545 consecutive patients with BPH undergoing prostatectomy at 3 medical centers in Israel between 1991 and 1992. Repeated structured interviews preoperatively, and at 4 and 12 months postoperatively were performed, including 6 quality of life questionnaires evaluating BPH specific (symptom severity and symptom effect) and generic (activity, independence, mental health and health perception) parameters. In addition, the interviews consisted of socio-demographic data elements. Clinical details regarding severity of prostatic disease and co-morbidity were obtained from the medical charts. RESULTS: We found a correlation between postoperative change in symptom effect and in generic quality of life measures (r 0.11 to 0.20, p < 0.04). The postoperative decrease in the mean symptom effect score was 56% and 52% for severe and moderate preoperative levels, respectively. There was no decrease in the mean symptom effect score for the mild preoperative level (18% of these patients had postoperative deterioration). A secondary operation, and the combination of diabetes mellitus and poor activity level were risk factors for lack of improvement in patients with moderate preoperative symptom effects. We found that the impact of open prostatectomy on quality of life was similar to that of the closed technique after adjustment for patient attributes, except for those with an indwelling urinary catheter in whom an open operation was advantageous. CONCLUSIONS: In patients with BPH and mild symptom effects, and in subgroups of patients with moderate symptom effects surgery should not be recommended. Based on short-term measures of quality of life there is no justification for a preference between open and closed operations. PMID- 7490832 TI - Do reported micturition symptoms on the American Urological Association Questionnaire correlate with 24-hour home uroflowmetry recordings? AB - PURPOSE: Patient subjective descriptions of micturition habits using the American Urological Association (AUA) symptom index were compared with uroflowmetry recordings obtained during a 24-hour period. MATERIALS AND METHODS: In a prospective study, the AUA questionnaire was given twice to 42 men. All micturitions during a typical 24-hour period were continuously recorded by a home uroflowmetry system. Daytime frequency, strength and intermittency of the stream, and nocturia were calculated from the recorded data, which were correlated with the matching questionnaire answers. RESULTS: Except for nocturia, there was no correlation between the answers and recorded findings. CONCLUSIONS: Men are unable to quantify correctly their own clinical status. The study results cast doubt on the validity of the self-administered AUA survey to reflect clinical micturition. PMID- 7490833 TI - Commentary on genetic prognostic markers for transitional cell carcinoma of the bladder: from microscopes to molecules. PMID- 7490834 TI - Correlation between the International Prostatic Symptom Score and a pressure-flow study in the evaluation of symptomatic benign prostatic hyperplasia. AB - PURPOSE: We examined the relationship between the International Prostatic Symptom Score (I-PSS) and the occurrence of bladder outlet obstruction caused by benign prostatic hyperplasia (BPH). MATERIALS AND METHODS: The American Urological Association developed a questionnaire to quantify the severity of symptoms resulting from BPH. A further question relating the impact of BPH to the quality of life was subsequently added. This questionnaire has been adopted by the World Health Organization and is known as the I-PSS. There are 4 questions related to obstructive symptoms and 3 related to irritative symptoms. Scores of 0 to 7, 8 to 19 and 20 to 35 represent mild, moderate and severe symptoms, respectively. During an 18-month interval the I-PSS questionnaire was administered to 258 patients 50 to 81 years old (mean age 63 years) with BPH. A pressure-flow study was used to determine the presence of bladder outlet obstruction in 227 patients. Based on the scores, the patients were divided into 31 with mild, 116 with moderate and 111 with severe obstruction. The pressure-flow study was not conducted on patients with a mild symptom score. RESULTS: Of the patients with a severe symptom score 92 (82.9%) had bladder outlet obstruction, compared to 62 (53.4%) with a moderate symptom score. Statistical analysis (Pearson chi-square test) showed that there was a significant positive correlation between the symptoms and the presence of bladder outlet obstruction. Thus, when the I-PSS was greater than 28, the probability of bladder outlet obstruction was more than 0.91. Stratification of the results according to the obstructive (0 to 20) and irritative (0 to 15) symptoms of the I-PSS yielded a significant positive correlation between obstructive symptoms and the presence of bladder outlet obstruction. Thus, when the obstructive symptom score was greater than 15, the probability of bladder outlet obstruction was greater than 0.91. CONCLUSIONS: When the total I-PSS is greater than 28 or the obstructive symptom score is greater than 15, a pressure-flow study must be avoided. PMID- 7490835 TI - Fine-tuning the indication for prostatectomy. PMID- 7490836 TI - Endoscopic evaluation and treatment of anastomotic strictures after radical retropubic prostatectomy. AB - PURPOSE: We describe endoscopic findings and treatment outcome in 17 men who presented with a symptomatic anastomotic stricture after radical prostatectomy. MATERIALS AND METHODS: Endoscopic evaluation revealed an immature stricture in 6 men, which was treated by initial dilation and subsequent cold-knife urethrotomy. The 11 men with a mature anastomotic stricture were treated by initial cold-knife urethrotomy (10) or dilation followed by urethrotomy (1). RESULTS: There were no long-term complications from treatment, which was successful in 15 of the 17 men (88%). CONCLUSIONS: Initial dilation with subsequent urethrotomy for immature or initial urethrotomy for mature anastomotic strictures is a safe and effective treatment plan. PMID- 7490837 TI - Flutamide hepatotoxicity. AB - PURPOSE: Observed and expected reporting rates were compared in patients who died or were hospitalized due to hepatotoxicity associated with the use of flutamide. MATERIALS AND METHODS: Case series were submitted to the MedWatch Spontaneous Reporting System of the Food and Drug Administration. Reporting rates for serious hepatotoxicity due to flutamide were calculated and compared to rates for hospitalized patients with acute idiopathic hepatitis in the medical literature. RESULTS: After the marketing of flutamide in the United States, between February 1989 and December 1994 the Food and Drug Administration received reports of 20 patients who died and 26 who were hospitalized for hepatotoxicity due to flutamide. The rate of approximately 3 per 10,000 flutamide users exceeds by 10 fold or more the expected rate of hospitalizations for acute noninfectious liver injury of 2.5 per 100,000 men 65 years and older. Autopsies in 6 cases revealed marked to massive hepatic necrosis as the predominant feature. CONCLUSIONS: Flutamide is a potent hepatotoxin in certain patients. Serial blood aminotransferase levels should be monitored during the first few months of flutamide treatment. Before beginning use of this drug patients should be instructed to report immediately to physicians any episodes of nausea, vomiting, fatigue and jaundice so that flutamide can be promptly discontinued to avoid progression of possible liver injury. PMID- 7490838 TI - Biochemical and pathological effects of 8 months of neoadjuvant androgen withdrawal therapy before radical prostatectomy in patients with clinically confined prostate cancer. AB - PURPOSE: A prospective, nonrandomized trial was initiated to determine the duration of neoadjuvant therapy required for prostate specific antigen (PSA) to reach its nadir, evaluate the ability of an ultrasensitive assay to measure decreases in PSA less than 0.2 microgram./l., and characterize the effects of 8 months of neoadjuvant therapy on pathological stage, positive margin rates, proliferation and tumor marker immuno-staining. MATERIALS AND METHODS: We evaluated 50 patients with clinically localized prostate cancer treated by 8 months of reversible androgen ablation before radical prostatectomy. Serum PSA and testosterone levels were measured monthly. RESULTS: Serum PSA decreased by 84% after 1 month and by a further 52% between 3 and 8 months. Using an ultrasensitive assay, serum PSA decreased to undetectable levels (less than 0.1 microgram./l.) or reached its nadir in 22% of the cases after 3 months, 42% after 5 months and 84% after 8 months. Overall, the positive margin rate was 4%. Of the cases 68% were organ-confined and 24% were specimen-confined. The positive margin rate was not increased after reevaluation with cytokeratin, PSA and prostatic acid phosphatase immuno-staining but of 4 cases initially staged as P0 on hematoxylin and eosin evaluation 2 had microscopic foci of cancer with prostatic acid phosphatase staining. Immuno-staining with the proliferation markers proliferation cell nuclear antigen and Ki-67 showed decreased staining in surgical specimens relative to pretreatment needle biopsy specimens, which suggests that outgrowth of androgen independent clones does not develop during prolonged neoadjuvant therapy. CONCLUSIONS: Eight months of neoadjuvant androgen withdrawal therapy results in low positive margin rates and PSA nadir levels. The initial rapid decrease in PSA results from cessation of androgen regulated PSA synthesis and apoptosis, while the ongoing slower decrease reflects decreasing tumor volume. PMID- 7490839 TI - A better prognosis for obese men with prostate cancer. AB - PURPOSE: The possibility was investigated whether obesity is associated with a favorable prognosis in men with nonstage A prostatic cancer independent of other risk factors. MATERIALS AND METHODS: A total of 235 men with nonstage A prostatic cancer diagnosed between 1983 and 1990 at a community hospital was evaluated by tumor grade and stage, degree of obesity, smoking habits, age, year of diagnosis and survival. RESULTS: In comparison to tumors of less obese men those in men who were at least 10% overweight were more often stage B or C than D (odds ratio 2.3, 95% confidence interval 1.4 to 4.9). Smoking abstinence, patient age older than 70 years and low Gleason scores were also independently associated with less advanced tumors. The lower frequency of advanced stage tumors among those of obese men was most prominent for stage D2 disease in nonsmokers, progressively decreasing from 24% of 29 nonsmokers who had been underweight for many years before diagnosis to 8% of 24 who had been greatly overweight for many years (p < 0.05). Tumor specific mortality was also lower in obese men and nonsmokers independent of tumor stage and grade with 5-year tumor specific mortality rates of 10% in obese nonsmokers, 27% in nonobese nonsmokers, 36% in obese smokers and 43% in nonobese smokers. These differences reflected a combination of obesity related differences in stage distribution and a more favorable prognosis for obese men among patients with tumors of similar stages. CONCLUSIONS: These observations suggest inhibition of prostate cancer growth and metastasis by increased endogenous estrogen, decreased endogenous testosterone or other systemic changes characteristic of obesity. PMID- 7490840 TI - Prostate cancer. PMID- 7490841 TI - Strategy for repeat biopsy of patients with prostatic intraepithelial neoplasia detected by prostate needle biopsy. AB - PURPOSE: We evaluated the strategy for repeat biopsy of patients with prostatic intraepithelial neoplasia without concurrent carcinoma detected on prostate needle biopsy. MATERIALS AND METHODS: Of 1,275 consecutive patients undergoing prostate needle biopsy 61 were identified with prostatic intraepithelial neoplasia but without concurrent prostate carcinoma. Of the 61 patients 53 had undergone repeat biopsy. The medical records, transrectal ultrasound, and operative and pathological reports of these patients were reviewed. RESULTS: Repeat biopsy was done in 53 patients with prostatic intraepithelial neoplasia, yielding carcinoma in 15, prostatic intraepithelial neoplasia without carcinoma in 8 and benign tissue in 30. The yield of carcinoma from repeat biopsy of a prostatic intraepithelial neoplasia site was 8.3% (7 of 84 sites). A total of 18 sites of carcinoma was detected by repeat biopsy of a previous random biopsy site (8), a prostatic intraepithelial neoplasia site only (5), a transrectal ultrasound nodule (3), a palpable nodule and prostatic intraepithelial neoplasia site (1), and a transrectal ultrasound nodule and prostatic intraepithelial neoplasia site (1). Carcinoma was as frequently detected by repeat biopsy of a prostatic intraepithelial neoplasia site (6 patients) as by random repeat biopsy (6 patients). CONCLUSIONS: Repeat prostate needle biopsy of patients with prostatic intraepithelial neoplasia should include random repeat biopsy and repeat biopsy of transrectal ultrasound abnormalities as well as previous sites of prostatic intraepithelial neoplasia. PMID- 7490842 TI - Limitations of serum prostate specific antigen in predicting peripheral and transition zone cancer volumes as measured by correlation coefficients. AB - PURPOSE: We reexamined the relationship between preoperative serum prostate specific antigen (PSA) and prostate cancer volume in 290 patients who underwent radical prostatectomy. MATERIALS AND METHODS: Serum samples from 290 consecutive patients were remeasured with the automated monoclonal-monoclonal Tosoh AIA-600 assay. These values were correlated with individual cancer volume by measuring Pearson correlation coefficients (r). RESULTS: Cancer was noted in the transition zone in 31 patients and in the peripheral zone in 259. Of the peripheral zone cancers 133 (51.4%) were organ confined and 126 (48.6%) were nonorgan confined, including 12 (9.5%) with histologically confirmed lymph node metastasis (stage D1). The 259 peripheral zone cancers had a correlation coefficient with PSA (r = 0.499, p < 0.0001). After distributing the 259 cases into cancer volume groups we found a large overlap in mean preoperative serum PSA, including 65 with 50% or greater Gleason grade 4 or 5 disease (r = 0.508). The correlation coefficients of cancer volume with PSA in 133 organ confined cancers, 114 nonorgan confined cancers without lymph node metastases and 12 nonorgan confined cancers with positive lymph nodes were 0.382, 0.438 and 0.363, respectively. The 31 transition zone cancers showed a correlation coefficient with PSA (r = 0.81). After excluding 2 cases with extreme PSA and cancer volume the correlation coefficient decreased (r = 0.077). CONCLUSIONS: Even when remeasured with an automated monoclonal-monoclonal assay serum PSA alone is unable to predict preoperatively cancer volume or distinguish between organ and nonorgan confined cancer in peripheral and transition zone tumors of the prostate. PMID- 7490843 TI - Molecular detection of prostate epithelial cells from the surgical field and peripheral circulation during radical prostatectomy. AB - PURPOSE: Prostate cancer progression despite organ confined pathological assessment has been reported in a variable number of men after radical retropubic prostatectomy. To study this phenomenon, we used the prostate specific antigen (PSA) reverse transcriptase-polymerase chain reaction assay. MATERIALS AND METHODS: We prospectively assayed the peripheral venous blood before, during and after surgical manipulation as well as the intraoperative field blood for PSA reverse transcriptase-polymerase chain reaction-positive cells in 22 men undergoing radical retropubic prostatectomy. RESULTS: PSA reverse transcriptase polymerase chain reaction-positive cells were identified in 20 of the 22 operative field samples (91%) and 4 of 16 (25%) had evidence of intraoperative hematogenous dissemination (p = 0.046). No significant association was identified among Gleason score, pathological stage and the PSA reverse transcriptase polymerase chain reaction result. CONCLUSIONS: Our results suggest that tumor cell spillage and less frequently hematogenous dissemination may be associated with operative manipulation of the prostate during radical retropubic prostatectomy and may potentially represent mechanisms of failure after radical retropubic prostatectomy. PMID- 7490844 TI - Prostate cancer--improvements in detection and diagnosis. PMID- 7490845 TI - Is stage pT4a (D1) reliable in assessing transitional cell carcinoma involvement of the prostate in patients with a concurrent bladder cancer? A necessary distinction for contiguous or noncontiguous involvement. AB - PURPOSE: A series of patients with concurrent transitional cell carcinoma involvement of the prostate and bladder is reviewed to define the impact of prostate involvement pathways and the degree of prostate invasion on survival rate. MATERIALS AND METHODS: A total of 72 patients who underwent radical cystectomy for pathological stage pT4a (D1) cancer was divided into contiguous- stage pT4a, transitional cell carcinoma of the bladder extended into the prostate through the bladder wall and noncontiguous--stage pT4a simultaneous transitional cell carcinoma of the prostate and bladder carcinoma that did not directly infiltrate into the prostate through the bladder wall. In the latter group the degree of prostate invasion was classified as urethral mucosal involvement, ductal/acinar involvement, stromal invasion and extracapsular extension. The survival rate was estimated by the Kaplan-Meier and Cox proportional hazards methods. Comparisons between curves were performed by univariate log rank and multivariate L-ratio tests. RESULTS: The overall 5-year survival rate for stage pT4a was 21.5% (median followup 64 months). Furthermore, 46% and 7% of patients in noncontiguous and contiguous pT4a groups, respectively, were estimated to be alive (p < 0.000). Those with positive nodes experienced a poor outcome in both groups. Of patients with noncontiguous pT4a stage 100% with urethral mucosal involvement, 50% with ductal/acinar involvement and 40% with stromal invasion were estimated to be alive. The major prognostic factors were bladder tumor stage, nodal involvement and degree of prostate invasion. CONCLUSIONS: The invasion pathways of the prostate in patients with transitional cell bladder carcinoma have a statistically significant prognostic role. Contiguous and noncontiguous involvements are 2 distinct clinicopathological features and they should not be included in the same stage. In the noncontiguous stage pT4a group bladder and prostate transitional cell carcinoma should be separately staged, and prostate involvement also should be staged according to invasion degree. PMID- 7490846 TI - Determining the origin of hematuria by immunocytochemical staining of erythrocytes in urine for Tamm-Horsfall protein. AB - PURPOSE: We analyzed the feasibility of immunocytochemical staining of urinary erythrocytes for Tamm-Horsfall protein to differentiate renal from nonrenal hematuria. MATERIALS AND METHODS: Urine samples collected from 74 patients with hematuria were evaluated. Erythrocytes in urine were inspected morphologically and the same samples were immunocytochemically stained with antibody against human Tamm-Horsfall protein. RESULTS: Erythrocytes in urine were clearly stained for Tamm-Horsfall protein in renal hematuria. Tamm-Horsfall protein immunocytochemistry was more consistent with clinical diagnosis than morphology. CONCLUSIONS: Immunocytochemical staining of urine erythrocytes for Tamm-Horsfall protein appears to be a reliable tool to differentiate renal from nonrenal hematuria. PMID- 7490847 TI - Cryotherapy for renal cell carcinoma and angiomyolipoma. AB - PURPOSE: We determined the feasibility of renal cryotherapy in humans. MATERIALS AND METHODS: Two patients with tumors in a solitary kidney were treated with renal cryotherapy. RESULTS: Both patients tolerated renal cryotherapy well with no change in renal function and no postoperative complications. CONCLUSIONS: Cryoablation of select renal lesions can be performed safely with minimal loss of renal function. PMID- 7490848 TI - Combined endoscopic closure of rectourethral fistula. AB - PURPOSE: Rectourethral fistula formation is a rare complication of radical prostatectomy and other pelvic surgical procedures. MATERIALS AND METHODS: In 2 patients endoscopic closure of the fistula was performed by combined transrectal endoscopic excision of the fistula, endoscopic suture, and simultaneous transurethral fulguration and fibrin application. RESULTS: This combined approach resulted in closure of the fistula in both patients as proved by rectoscopy, retrograde urethrography and disappearance of all clinical symptoms. The latest postoperative followup was 18 months. CONCLUSIONS: Any open procedure to correct a rectourethral fistula is considered major surgery. Therefore, the minimally invasive approach described should be attempted first in patients with a small rectourethral fistula. PMID- 7490849 TI - Quality of life after a Marshall-Marchetti-Krantz procedure for stress urinary incontinence. AB - PURPOSE: We assessed the long-term results of the Marshall-Marchetti-Krantz procedure. MATERIALS AND METHODS: A total of 151 case records was reviewed and a questionnaire was mailed to patients to estimate the success rate and patient satisfaction. RESULTS: The response rate was 67.3% (101 replies). The overall success rate was 72.8% (mean followup 51.5 months). The success rates in patients with irritative symptoms and pure stress incontinence were 65% and 82.6%, respectively. The procedure was equally effective for recurrent stress incontinence. CONCLUSIONS: The procedure is characterized by a high success rate 5 years postoperatively. The method of review, based on a mailed questionnaire, appears to be more objective than conventional reviews but further improvements may increase the accuracy. PMID- 7490850 TI - Therapy of locally recurrent renal cell carcinoma after nephrectomy. AB - PURPOSE: We evaluated the outcome of patients with locally recurrent renal cell carcinoma treated at our university. MATERIALS AND METHODS: We retrospectively analyzed 16 cases of locally recurrent renal cell carcinoma in the renal fossa treated with surgical resection alone or in combination with biological therapy. RESULTS: Complete resection was possible in 15 patients of whom 3 had positive surgical margins. Of the 12 patients with negative margins 6 are free of disease while the 3 with positive margins had involvement of a remaining ipsilateral adrenal gland and distant metastasis. Of all 16 patients 12 are alive a median of 23.5 months after the diagnosis of locally recurrent renal cell carcinoma. Of those treated with the combination of biological therapy and surgery 50% have no evidence of disease compared to 25% of those treated with surgery alone. CONCLUSIONS: Long-term survival can be achieved with an aggressive surgical approach. Incomplete resection or positive surgical margins are associated with a high risk of local or distant failure, and combined treatment with immunotherapy and surgery may offer a benefit compared to surgery alone. PMID- 7490851 TI - The role of urodynamics in the evaluation of voiding dysfunction in men after cerebrovascular accident. AB - PURPOSE: The etiology of voiding dysfunction was determined in men after a cerebrovascular accident who were at risk for obstructive uropathy to evaluate whether the cause of voiding dysfunction could be predicted by the type (obstructive or irritative) or onset of symptoms. MATERIALS AND METHODS: We evaluated 38 men with complaints of voiding dysfunction following a cerebrovascular accident. All patients were of the age when bladder outlet obstruction secondary to benign prostatic hyperplasia would otherwise be prevalent. After a comprehensive history and physical examination, all patients underwent multichannel urodynamic studies at a medium fill rate (20 to 50 ml. per minute). Findings were classified by the Abrams-Griffiths nomogram as obstruction, no obstruction or equivocal. RESULTS: Mean patient age was 70 years (range 54 to 87). Patients were grouped according to the presenting voiding complaints (purely irritative in 42%, purely obstructive in 34% or mixed in 24%). In 34 patients (89%) the onset of symptoms paralleled the occurrence of the cerebrovascular accident. Detrusor hyperreflexia was noted in 82% of the patients. There was no statistically significant difference in the occurrence of detrusor hyperreflexia among the 3 symptom groups (Fisher's exact test). Pressure flow analysis clearly showed obstruction in 24 patients (63%), no obstruction in 9 (24%) and equivocal results in 5 (13%) according to the nomogram. There was no statistically significant difference in the incidence of obstruction among the 3 symptom groups (Fisher's exact test). CONCLUSIONS: Presenting symptoms did not predict the urodynamic findings of bladder outlet obstruction or detrusor hyperreflexia. The significant incidence of onset of symptoms after stroke suggests that the cerebrovascular accident induced voiding dysfunction in the face of preexisting bladder outlet obstruction may exacerbate the symptoms of the latter condition or vice versa. PMID- 7490852 TI - Pressure-to-cross-sectional area relationships in the proximal urethra of men with bladder outlet obstruction. AB - PURPOSE: Related values of pressure and cross-sectional area in the proximal urethra were measured in patients with bladder outlet obstruction. Urethral opening pressure and elastance (the inverse of compliance) were estimated. MATERIALS AND METHODS: We studied 15 men with standard urodynamic examinations. The pressure-to-cross-sectional area relationship in the prostatic urethra was determined using a special probe. RESULTS: Elastance varied significantly along the studied portion of the urethra, with higher values found in the sphincter area. The estimated urethral opening pressure appeared high compared to that in unobstructed cases and without variation along the prostatic urethra. CONCLUSIONS: The most important effect of prostatic obstruction appears to be the increased urethral opening pressure. PMID- 7490853 TI - Morphological and functional factors predicting bladder deterioration after spinal cord injury. AB - PURPOSE: We investigated factors predictive of morphological and functional deterioration of the bladder in patients with spinal cord injury. MATERIALS AND METHODS: Urological evaluation, including excretory urography and urodynamic studies, was performed once a year in 66 patients. Bladder compliance was used as an index of bladder function. RESULTS: Mean age of the patients with worsening bladder form and function was significantly greater than that of those with no change. In patients with worsening bladder function intermittent catheterization was performed less frequently and mean catheterization volume was greater compared to those with improvement and no change. Catheterization was less frequent and at long intervals in patients with a high urethral closure pressure. CONCLUSIONS: High storage pressure due to a high urethral closure pressure is believed to cause deterioration of bladder form and function. PMID- 7490854 TI - The evolving importance of neurourology and urodynamics. PMID- 7490856 TI - Correlation of Valsalva leak point pressure with subjective degree of stress urinary incontinence in women. AB - PURPOSE: We correlated symptoms of stress urinary incontinence in women with intrinsic urethral function, as measured by Valsalva leak point pressure. In addition, we assessed the effects of urethral hypermobility, baseline resting abdominal pressure, patient age and menopausal status on Valsalva leak point pressure. MATERIALS AND METHODS: We evaluated 64 consecutive women (mean age 50 years, range 20 to 79) with complaints of stress urinary incontinence. Initially the patient history was obtained and the subjective degree of stress urinary incontinence was graded as 1, 2 or 3 according to the SEAPI-QMN classification. Multichannel video urodynamics were then performed. At a volume of 150 to 200 ml. (or half bladder capacity when functional capacity was less than 150 ml.) filling was stopped. Resting, coughing and progressive Valsalva maneuvers were performed to determine Valsalva leak point pressure and the presence of urethral hypermobility. Valsalva leak point pressure was defined as the minimum total vesical pressure required to cause urinary incontinence in the absence of a detrusor contraction. Urodynamics were completed and detrusor instability or stress induced instability was noted. RESULTS: Of the 64 women 52 (81.3%) had stress urinary incontinence. The exact test for trend demonstrated a statistically significant difference in the number of patients with a Valsalva leak point pressure of 90 cm. water or less (p = 0.0002) and 60 cm. water or less (p = 0.0002) among the 3 symptom groups. There was no correlation between Valsalva leak point pressure and resting vesical (which equals abdominal) pressure, patient age or menopausal status, or urge incontinence or detrusor instability among the 3 groups. CONCLUSIONS: The subjective degree of stress urinary incontinence can predict intrinsic urethral function as measured by Valsalva leak point pressure. Higher grades of stress urinary incontinence have a higher likelihood of a low Valsalva leak point pressure. Many women with grade 2 or 3 stress urinary incontinence have a Valsalva leak point pressure of 90 cm. water or less despite urethral hypermobility and they may have a component of intrinsic urethral deficiency. PMID- 7490855 TI - Reappraisal of endoscopic sphincterotomy for post-traumatic neurogenic bladder: a prospective study. AB - PURPOSE: We assessed the results of endoscopic sphincterotomy in patients with detrusor-sphincter dyssynergia secondary to spinal cord injury. MATERIALS AND METHODS: A prospective study was done of 92 patients with a mean followup of 20.6 months. RESULTS: The morbidity rate was 10.9%. Results demonstrated objective (assessed by the decrease in voiding pressure) and subjective (expressed by the patient) improvement in quality of micturition in 83.7% and 73% of the cases, respectively. Subjective autonomic dysreflexia resolved in 93.2% of the cases. CONCLUSIONS: Sphincterotomy is a useful and effective therapeutic method for neurogenic bladder when all available pharmacological treatments and clean intermittent catheterization have failed. PMID- 7490857 TI - Incontinence--lower urinary tract dysfunction and urodynamic evaluations. PMID- 7490858 TI - Primary squamous cell carcinoma of the ureter and squamous adenocarcinoma of the renal pelvis: 2 case reports. PMID- 7490859 TI - Re: Indwelling ureteral stents as definitive therapy for distal ureteral calculi. PMID- 7490860 TI - Re: Indwelling ureteral stents as definitive therapy for distal ureteral calculi. PMID- 7490861 TI - Re: Extensive urodynamic investigation: interaction among diuresis, detrusor instability, urethral relaxation, incontinence and complaints in women with a history of urge incontinence. PMID- 7490862 TI - Re: Fournier's gangrene as the presenting sign of an undiagnosed human immunodeficiency virus infection. PMID- 7490863 TI - Re: Histopathological and cytopathological correlations of percutaneous testis biopsy and open testis biopsy in infertile men. PMID- 7490864 TI - Re: Automated prostate volume determination with ultrasonographic imaging. PMID- 7490865 TI - Re: Editorial: benign prostatic hyperplasia. PMID- 7490866 TI - Re: Potency-sparing radical retropubic prostatectomy: a simplified anatomical approach. PMID- 7490868 TI - Re: The clinical usefulness of prostate specific antigen: update 1994. PMID- 7490867 TI - Re: The clinical usefulness of prostate specific antigen: update 1994. PMID- 7490870 TI - Re: Nontraumatic elevation of prostate specific antigen following cardiac surgery and extracorporeal cardiopulmonary bypass. PMID- 7490869 TI - Re: The clinical usefulness of prostate specific antigen: update 1994. PMID- 7490871 TI - Re: Eliminating the need for bilateral pelvic lymphadenectomy in select patients with prostate cancer. PMID- 7490872 TI - Re: Long-term survival and mortality in prostate cancer treated with noncurative intent. PMID- 7490873 TI - The effect of finasteride on prostate specific antigen: review of available data. AB - PURPOSE: We reviewed the available data on the effect of finasteride on serum levels of prostate specific antigen (PSA), PSA velocity and PSA density in men with benign prostatic hyperplasia (BPH) and prostate cancer. MATERIALS AND METHODS: To our knowledge all previously published analyses of PSA data from clinical trials of finasteride therapy for BPH and prostate cancer are reviewed. RESULTS: The normal reference range of serum PSA levels in men with BPH and no evidence of prostate cancer who were treated with finasteride for 6 months or longer is half that in untreated men with BPH. The percent by which serum levels of PSA are suppressed after 6 months of treatment for prostate cancer does not tend to be greater than that for BPH. CONCLUSIONS: To interpret serum PSA levels in men with BPH treated with finasteride for 6 months or longer, the serum PSA level should be multiplied by 2 and compared to either age-independent or age specific upper limits of normal for serum PSA in untreated men with BPH. On the basis of limited data the sensitivity and specificity of this approach appear to be similar to those of the corresponding PSA limits in untreated men with BPH. PMID- 7490875 TI - Complete penile disassembly for epispadias repair: the Mitchell technique. AB - PURPOSE: We report a new technique for epispadias repair, which relies on the unique blood supply of the corpus cavernosum and glans. The epispadiac phallus is completely disassembled into 3 discrete components: the urethral plate, and the right and left hemicorporeal glandular bodies. MATERIALS AND METHODS: Ten boys 10 months to 17 years old presented for repair of epispadias between 1990 and 1994 (6 primary and 4 secondary repair). One patient underwent bladder exstrophy closure at the time of epispadias surgery. Two patients underwent prior para exstrophy flap procedures and neither had fistulas. RESULTS: Followup (8 to 57 months) revealed a conical glans in 8 patients, straight shaft upward in 1 and ventral orthotopic meatus in 7. In 2 patients who underwent secondary repair 3 fistulas formed that were repaired on an outpatient basis. All patients are potent. CONCLUSIONS: Complete disassembly allows tubularization and ventralization of the entire distal urethra; makes glans and urethral repair independent; separates the 2 corporeal glandular bodies, permitting easier and more complete release of the rotation contributing to dorsal chordee, and improves exposure for corporotomy or dermal grafts. PMID- 7490876 TI - Bladder wall pedicle wraparound sling for neurogenic urinary incontinence in children. AB - PURPOSE: To improve the results of current bladder neck reconstruction a modification was developed to provide tapering, circumferential compression and suspension of the bladder neck. MATERIALS AND METHODS: Nine female and 15 male patients with spinal dysraphism and incontinence, despite intermittent catheterization and pharmacotherapy, and low outlet resistance underwent surgery. The procedure involves wrapping a pedicle strip of anterior bladder wall around the bladder neck and suspending it to the pubic symphysis. RESULTS: All 9 girls are completely continent, whereas 10 boys are dry and 5 have stress incontinence after 9 to 14 months of followup. Subjective symptoms correlate with changes in leak point pressure. CONCLUSIONS: This technique is an efficacious addition to bladder augmentation in female patients with poor compliance and low outlet resistance. PMID- 7490874 TI - Splenectomy complicating left nephrectomy. AB - PURPOSE: We attempted to clarify the details of incidental splenectomy complicating left nephrectomy. MATERIALS AND METHODS: We reviewed the literature and operations involving splenectomy performed during left nephrectomy between 1984 and 1994 at our university. Factors reviewed included patient characteristics, renal pathology, mechanisms of injury, blood transfusions and postoperative complications. RESULTS: Of the 418 left nephrectomies 18 (4.3%) resulted in splenectomy via a transperitoneal approach. Patients with a large or upper pole renal lesion, malignancy or advanced age are increasingly likely to undergo unanticipated splenectomy. CONCLUSIONS: Our results, combined with recommendations from the Centers for Disease Control and Prevention, suggest that all patients older than 65 years undergoing left transperitoneal nephrectomy or those at increased risk for splenic injury should receive preoperative pneumococcal vaccination. PMID- 7490877 TI - Splenogonadal fusion: report of 2 cases and review of the literature. AB - PURPOSE: We reviewed the presentation and clinical findings in an unusual scrotal mass seen in children. MATERIALS AND METHODS: This review involves a detailed description of 2 cases of splenogonadal fusion, and a complete survey and assessment of the published literature on this subject. RESULTS: A detailed assessment of 137 cases (135 previously reported, 2 added) of splenogonadal fusion was undertaken. Splenogonadal fusion can be classified as either continuous or discontinuous type. The disorder was rarely recognized before surgery. CONCLUSIONS: In 37% of reported patients orchiectomy was performed because of suspicion that the lesion represented a primary neoplasm. However, orchiectomy is not necessary, since the splenic tissue can be dissected safely off of the tunica albuginea. Therefore, an awareness of the possibility that this anomaly may exist can avoid an unnecessary orchiectomy. PMID- 7490878 TI - Incidence of apoptosis, cell proliferation and bcl-2 expression in transitional cell carcinoma of the bladder: association with tumor progression. AB - PURPOSE: Apoptosis is the distinctive form of programmed cell death that complements cell proliferation in maintaining normal tissue homeostasis. The significance of constitutive apoptosis in the development and progression of transitional cell carcinoma of the bladder has yet to be investigated. In the present study, the incidence of baseline apoptosis and the expression of 2 genes regulating this molecular process, bcl-2 and TGF-beta 1, as well as the level of cell proliferation, were examined by an intensive immunohistochemical analysis in normal bladder and bladder cancer specimens. MATERIALS AND METHODS: Apoptosis was detected by in situ end-labeling of fragmented DNA using the terminal deoxynucleotidyl transferase reaction in 45 paraffin-embedded primary transitional cell carcinoma specimens, 9 metastatic lymph nodes and 5 normal bladder specimens. The proliferation status of the tumor cells among the same bladder cancer specimens was evaluated by using a monoclonal antibody that recognizes the proliferation-associated nuclear antigen, Ki-67. RESULTS: The apoptotic index of normal transitional epithelium (0.06%) was significantly lower than that of all grades of transitional bladder carcinoma (p = 0.006). Although the apoptotic index of transitional carcinomas increased with increasing grade, this difference failed to achieve statistical significance, ranging from 0.54 +/- .23% in grade I to 1.24 +/- .77% in grade III. The proliferative index, as determined by Ki-67 positivity, also increased with increasing grades of tumor (12.8 +/- 8.4% in grade I to 22.6 +/- 15.2% in grade III) and was significantly greater than in normal urothelium (0.64 +/- 0.52%, p = 0.003). Bcl-2 expression was significantly lower in the normal transitional epithelium and in the well and moderately differentiated tumors (grades I-II) when compared with poorly differentiated (grade III) tumors (p = .004). The incidence of bcl-2 expression in all bladder specimens analyzed was uniformly low (< 5.3%). Transforming growth factor-beta 1 expression was not detected in any of the normal bladder specimens, primary tumors, or metastatic lymph nodes analyzed. CONCLUSIONS: The present findings revealed that no statistically significant correlation exists between the frequency of apoptosis and the pathological stage of bladder tumors, while they clearly demonstrate a strong direct correlation between an increased rate of cell proliferation and bladder cancer progression. PMID- 7490879 TI - C-erbB-2 in bladder cancer: molecular biology, correlation with epidermal growth factor receptors and prognostic value. AB - PURPOSE: To study the c-erbB-2 oncogene in primary transitional cell bladder cancer. MATERIALS AND METHODS: Ninety-five patients with known clinical follow-up and epidermal growth factor receptor (EGFr) status were studied for expression of c-erbB-2 by immunostaining. Possible mechanisms underlying increased staining for c-erbB-2 protein were investigated by analyzing DNA and RNA encoding c-erbB-2. RESULTS: Strong positive staining for c-erbB-2 was detected in 20 (21%) tumors, with weaker staining in a further 13 (14%). There was no correlation between increased staining for c-erbB-2 and tumor stage, grade, or EGFr status. There was a low rate of amplification of the c-erbB-2 gene (1 of 24) on Southern blotting with a higher rate of elevated c-erbB-2 mRNA (4 of 44) with dot blot hybridization. For pT1 tumors, the rate of recurrence was higher for those tumors which were positive for c-erbB-2. CONCLUSIONS: c-erbB-2 oncoprotein is expressed by a significant proportion of transitional cell tumors of the bladder. In this study, the prognostic significance of c-erbB-2 expression appears limited. PMID- 7490880 TI - Effects of long-term cocaine exposure on spermatogenesis and fertility in peripubertal male rats. AB - PURPOSE: This study was conducted to investigate the effects of long-term administration of cocaine on spermatogenesis and fertility in adult male rats. MATERIALS AND METHODS: Thirty-day-old male Sprague-Dawley rats were given cocaine hydrochloride (15 mg./kg. body weight, corresponding to an average single dose for a heavy cocaine user) either daily or twice weekly (weekend group, cocaine given on Saturday and Sunday) and mated with pregnancy-proven female rats after 100 and 150 days of exposure to the drug. Pregnancy rates and litter birth weights were evaluated. Serum testosterone, follicle stimulating hormone and luteinizing hormone levels were measured in all adult rats. Morphologic analysis of the testis entailed the evaluation of quantitative and qualitative histologic parameters to assess the effect of cocaine on various stages of spermatogenesis. RESULTS: After 100 days of treatment, the rats receiving daily cocaine had a pregnancy rate of only 33% versus 86% for the controls (p < 0.05). In rats exposed to cocaine for 150 days the pregnancy rate was 50% compared with 100% for controls (p < 0.05). The birth weights of offspring from the group receiving daily cocaine was 10% less than that of controls (p < 0.05). The weight of the testis and epididymis was not affected by cocaine exposure. Morphometric analysis showed significant differences between the cocaine-treated groups (both the daily cocaine and twice weekly cocaine groups) and their respective controls. The mean diameter of seminiferous tubules in the daily and twice weekly cocaine groups was reduced when compared with their respective controls. These differences between treated groups and their controls were statistically significant (p < 0.05). Similarly the thickness of the germinal epithelium was less in the cocaine treated groups than in the controls (p < 0.05). Degenerating cells were more numerous in both daily and twice weekly cocaine groups than the controls. Furthermore, the number of step VII spermatids was reduced in both daily and twice weekly cocaine groups, a difference that was statistically significant (p < 0.05). CONCLUSION: Our findings demonstrate that chronic administration of cocaine to peripubertal male rats has a profound effect on their testicular function. Even with twice weekly administration there was a significant adverse effect on spermatogenesis although this was not manifested by diminished fertility in this group. These findings confirm that chronic administration of cocaine to male rats can have a deleterious effect on spermatogenesis and fertility. PMID- 7490881 TI - The effects of K+ channel blockers on the spontaneous electrical and contractile activity in the proximal renal pelvis of the guinea pig. AB - PURPOSE: The role of various K+ channel populations in determining the time course and amplitude of the spontaneous action potentials and contractions in the smooth muscle cells of the guinea pig renal pelvis were investigated by standard electrophysiological and tension recording techniques. MATERIALS AND METHODS: Electrical recordings in the proximal renal pelvis were used to demonstrate the presence of 3 cell populations based on the waveform of their Ca(2+)-dependent action potentials which we have termed "pacemaker," "intermediate" and "driven" action potentials. RESULTS: Blockade of large conductance (BK) or small conductance (SK) Ca(2+)-activated K+ channels with charybdotoxin (30 to 60 nM.) and tetraethylammonium (TEA) (0.5 to 2 mM.) or apamin (200 nM.) increased the duration of the action potentials recorded in "driven" cells. Tetraethylammonium (2 mM.) and 4-aminopyridine (2 mM.) increased the frequency of action potential discharge in both "driven" and "pacemaker" cells, as well as having a positive inotropic and chronotropic effect on the spontaneous contractions of the renal pelvis. In contrast, glibenclamide (0.1 to 1 microM.), Cs2+ (1 mM.) and Ba2+ (100 microM.) had little effect on either the contractile or electrical activity of the renal pelvis. CONCLUSION: It was concluded that the frequency of contraction in the proximal renal pelvis is determined by the frequency of action potential discharge in "pacemaker" cells. On the other hand, increases in the amplitude of pelvic contractions may well be correlated with membrane depolarization and/or increases in the duration of the action potentials recorded in "driven" cells. PMID- 7490882 TI - A potential use of a monoclonal antibody to placental alkaline phosphatase (PLAP) to detect lymph node metastases of seminoma. AB - PURPOSE: To evaluate the ability of monoclonal antibodies against alkaline phosphatase (AL-P) for tumor immunolocalization in nude mice. MATERIALS AND METHODS: Two monoclonal antibodies against placental alkaline phosphatase (PLAP) and liver alkaline phosphatase (LAP) were intravenously administered to nude mice bearing AL-P producing HeLa Hep 2 tumors. Sera and tumors from 65 patients with testicular germ cell tumors were examined for PLAP levels. RESULTS: Levels of PLAP or LAP in the HeLa xenografts were shown to be comparable to levels of PLAP in seminomas, including lymph node metastases, or nonseminomas, respectively. A biodistribution study showed that the percentage of the injected dose of I-125 labelled anti-PLAP in the xenografts was rather constant at around 7% until 7 days after injection, while the percentage of the injected dose of I-125 labelled anti-LAP MAb decreased with time as in other tissues which do not contain significant amounts of PLAP or LAP. On scintiscan, the xenografts in nude mice were distinctly visualized at 7 days after injection of anti-PLAP MAb. CONCLUSIONS: These data may support the potential use of the anti-PLAP MAb for the immunodetection of lymph node metastases of seminoma. PMID- 7490883 TI - Expression of the homeotic gene Hox-d13 in the developing and adult mouse prostate. AB - PURPOSE: To examine expression of the homeotic gene Hox-d13 in the developing mouse prostate. MATERIALS AND METHODS: Expression was assayed by Northern analysis, reverse-transcriptase polymerase chain reaction and in situ hybridization. RESULTS: Expression is present in the urogenital sinus in late gestation and is localized postnatally in the urethral epithelium and prostatic ducts. Expression continues throughout development and persists in the adult prostate. CONCLUSION: A restricted domain of Hox-d13 expression in the developing urogenital tract is consistent with the role of the Hox genes in specifying regional identity during development. Expression in the prostate postnatally suggests additional roles in prostate ductal morphogenesis and/or cell differentiation. PMID- 7490884 TI - Autocrine motility factor receptor as a possible urine marker for transitional cell carcinoma of the bladder. AB - PURPOSE: To determine whether autocrine motility factor receptor (AMFR) is detectable in the urine of patients with transitional cell carcinoma (TCC) of the bladder. MATERIALS AND METHODS: We assayed the urine of 89 patients with bladder pathology and 28 normal controls for AMFR. A monoclonal antibody to AMFR was used. RESULTS: All patients with muscle-invasive TCC tested positive for AMFR. Autocrine motility factor receptor was detectable for 80% of superficial tumors, with a correlation between AMFR and tumor grade. Seventy-five percent of control urines tested negative. CONCLUSIONS: Autocrine motility factor receptor is detectable in the urine of patients with TCC. Long-term follow-up and refinements in the assay should define the marker's utility for detection and prognosis. PMID- 7490885 TI - Acute spermatic cord torsion alters the microcirculation of the contralateral testis. AB - PURPOSE: The purpose of this study was to characterize the hemodynamic changes in the contralateral testis during acute spermatic cord torsion in anesthetized rats. MATERIALS AND METHODS: We used videomicroscopy to examine the microcirculation of the contralateral testis following acute torsion. Specifically, we examined the effect on vasomotion, a rhythmic dilation and constriction of the arterioles that is involved in fluid and nutrient exchange and modulation of local vascular resistance. In a separate set of experiments, blood flow in the contralateral internal spermatic artery was measured with an ultrasonic flow probe during acute torsion. RESULTS: Following 720 degrees torsion, the amplitude of vasomotion in the contralateral testis increased 121% (29.0 +/- 3.9% versus 13.0 +/- 1.7%) compared with controls. Blood flow in the contralateral internal spermatic artery decreased 43% after 2 hours' torsion. CONCLUSIONS: Acute spermatic cord torsion altered the microcirculation by increasing the amplitude of vasomotion and decreased total blood flow to the opposite testis. Because the hydraulic resistance of a blood vessel exhibiting vasomotion is always less than a vessel with the same average but static diameter, the observed microcirculatory changes may be an adaptive response to preserve local flow in the presence of decreased total flow. The long-term consequences of these changes in the microcirculation may affect testicular function and ultimately fertility. PMID- 7490886 TI - Role of spinal nitric oxide in the facilitation of the micturition reflex by bladder irritation. AB - PURPOSE: Nitric oxide (NO) is known to have an important transmitter function at peripheral synapses in the urogenital tract and has also been implicated in the transmission of nociceptive information in the spinal cord. The present study evaluated the role of NO in the central micturition reflex pathway. MATERIALS AND METHODS: We examined the effect of N-nitro-L-arginine methyl ester (L-NAME), an inhibitor of NO synthase, on micturition reflexes induced by continuous infusion of saline or 0.1% acetic acid (a noxious stimulus) into the bladder in urethane anesthetized female rats. Bladder and external urethral sphincter function were monitored with a continuous cystometrogram (CMG) and electromyography (EMG). RESULTS: Intrathecal injection of L-NAME (0.01 to 1 mumol.) did not significantly change the CMG or sphincter EMG during saline infusion. Infusion of acetic acid decreased the intercontraction interval (ICI), indicating a decrease in the volume threshold for inducing micturition. Subsequent intrathecal administration of L-NAME partially reversed the decreased ICI in a dose-dependent manner, but did not change the amplitude of bladder contractions: 0.01, 0.1 and 1 mumol. of L NAME produced increases of 25%, 31% and 56% in the ICI. D-NAME, the inactive stereoisomer had no effect. This effect of L-NAME was reversed by injection of L arginine (2 mumol. intrathecally) which, by itself, did not alter ICI during saline infusion or acetic acid infusion. CONCLUSIONS: These results indicate that: (1) spinal NO containing pathways do not play a role in the normal micturition reflex, (2) NO is involved at the spinal level in the facilitation of the micturition reflex by nociceptive bladder afferents activated by noxious chemical irritation of the bladder. PMID- 7490887 TI - Major histocompatibility complex class I and class II expression in renal cell carcinoma and modulation by interferon gamma. AB - PURPOSE: To determine the expression of MHC class I and II in human renal cancer. MATERIALS AND METHODS: We analyzed tissue sections from 22 primary and 28 metastatic renal cell carcinomas (RCC), as well as 31 established RCC cell lines. Tissue specimens from normal kidney and cell cultures of normal kidney epithelium were also studied. In addition, MHC antigen expression on RCC cell lines was assessed both before and after incubation with human recombinant interferon gamma (IFN-gamma). Antigen expression was determined by mixed hemadsorption, indirect immunofluorescence, fluorescence activated cell sorting (FACS) or immunoperoxidase staining using the monoclonal antibodies (mAbs) W6/32 (anti-MHC class I), mAbs NAMB-1 and BBM.1 (anti-beta-2 microglobulin), and mAbs L243 and 13 17 (anti-MHC class II) antibodies. Soluble beta-2 microglobulin in conditioned medium was measured by ELISA. RESULTS: Normal renal epithelial cells, both in vivo and in vitro, showed low level expression of class I antigens. Immunohistochemical staining for MHC class II was limited to some proximal tubular cells, while cultured renal tubular cells were uniformly class II negative. The tumor cell populations in all 22 primary and in 26 of 28 (93%) metastatic RC specimens consisted predominantly of class I positive cells. Half of the samples from primary and metastatic tumors were class II negative. Incubation of RCC cell lines with IFN-gamma enhanced the expression of MHC class I, beta-2 microglobulin and class II. The upregulation of MHC expression was time and dose dependent and associated with increased release of soluble beta-2 microglobulin. CONCLUSIONS: (i) Like normal kidney, virtually all primary human renal cell carcinomas express MHC class I antigens and retain this phenotype even during tumor progression and metastasis; (ii) class II expression on normal and RCC cells appears more limited but occurs frequently in both primary and metastatic lesions; and (iii) in most continuous RCC cell lines expression of MHC class I and II can effectively be stimulated by IFN-gamma. Since expression of MHC molecules might determine the immunogenicity of human RCC, its constitutive expression and augmentation could play an important role for the immunotherapy and prognosis of human renal cancer. PMID- 7490888 TI - Effect of two new polysaccharides on growth, agglomeration and zeta potential of calcium phosphate crystals. AB - PURPOSE: To study the effect of semisynthetic sulphated polysaccharides in the different calcium phosphate crystallization processes in vitro. MATERIALS AND METHODS: Crystallization of hydroxyapatite (HAP) and brushite (DCPD) in the presence and absence of 2 new semisynthetic sulphated polysaccharides (G871, G872) were defined by a constant composition technique, particle size analysis and zeta potential measurement. RESULTS: These polysaccharides demonstrated strong inhibitory effect on HAP and DCPD crystal growth and agglomeration. The increase of negative zeta potential values after addition of polysaccharides suggests the binding of these polysaccharides to HAP and DCPD crystals. CONCLUSION: We conclude that both G871 and G872 could be of potential use for calcium phosphate urolithiasis prevention in addition to their use for calcium oxalate. PMID- 7490889 TI - Renal cell carcinoma--local recurrence/splenic injury. PMID- 7490890 TI - Detrusor regeneration in the rat using porcine small intestinal submucosal grafts: functional innervation and receptor expression. AB - PURPOSE: To evaluate functional characteristics of regenerated bladder induced by small intestinal submucosa (SIS). MATERIALS AND METHODS: Strips from bladder regenerated from SIS and normal rat bladder were evaluated by in vitro muscle bath contractility studies. RESULTS: The present results indicate that SIS regenerated bladder 1) demonstrates contractile activity; 2) expresses muscarinic, purinergic and beta adrenergic receptors; and 3) exhibits functional cholinergic and purinergic innervation that is similar to the normal rat urinary bladder muscle. CONCLUSIONS: These functional characteristics of SIS-regenerated tissue demonstrated in the present study further support use of SIS material as a bladder augmentation material. PMID- 7490891 TI - Immunity to nerve growth factor prevents afferent plasticity following urinary bladder hypertrophy. AB - PURPOSE: The goal of this investigation was to examine the effect of immunity to nerve growth factor (NGF) on alterations in sensory nerves from the urinary bladder in the dorsal root ganglia (DRG) and their projections to the L6/S1 spinal cord following urethral obstruction in the rat. MATERIALS AND METHODS: Female Wistar rats were immunized to murine 2.5S NGF, then obstructed by partial urethral ligation for 6 weeks. Retrograde axonal tracing with FluoroGold and WGA HRP was used to measure areas of bladder DRG cells and afferent projections in the sacral spinal cord. Multiunit activity on bladder nerves allowed recording of micturition reflexes. Immunohistochemical staining for growth associated protein (GAP)-43 in the sacral parasympathetic nucleus (SPN) was used to assess potential growth or activity of axons in the spinal cord. Voiding frequencies were then measured in awake obstructed and NGF immune-obstructed rats. RESULTS: Immunity to NGF prevented obstruction-induced hypertrophy of DRG neurons, reduced retrograde axonal labeling of sacral afferent projections, eliminated enhancement of a spinal micturition reflex and abolished the increased GAP-43 expression in the SPN. Immunity to NGF prevented the urinary frequency that accompanies obstruction. CONCLUSIONS: Our results demonstrate that obstruction of the bladder elicits structural and functional plasticity in afferents as a result of ongoing neurotrophic interactions. Neurotrophic interactions offer a potential mechanism whereby symptoms and bladder hyperactivity develop with obstruction associated with benign prostatic hyperplasia. PMID- 7490892 TI - Ureteroscopic removal of mid and proximal ureteral calculi. AB - PURPOSE: Extracorporeal shock wave lithotripsy (ESWL) is considered standard therapy for most renal calculi as well as more than 80% of all upper urinary tract stones. The development of flexible and small diameter rigid ureteroscopes, and the increased availability of laser lithotriptors have made endoscopic treatment of ureteral calculi more successful and safe. This report represents our contemporary series of 80 single mid and proximal ureteral calculi treated endoscopically. MATERIALS AND METHODS: A total of 78 patients underwent 80 primary endoscopic procedures with 3 auxiliary ESWL and 2 endoscopic re treatments for 41 mid and 39 proximal ureteral calculi. Lithotripsy was performed for 55 of the 80 stones. RESULTS: Of the 80 stones 75 (94%) successfully cleared 1 month after 1 endoscopic procedure. The success rate was 38 of 39 (97%) for proximal calculi and 37 of 41 (90%) for mid ureteral stones. With a single re treatment overall success increased to 96%. If adjuvant ESWL was added, 100% success was attained. Currently all procedures are performed on an outpatient basis. Morbidity is low and there have been no major complications, such as stricture, renal loss or death. CONCLUSIONS: Our series demonstrates the high success rate that can be achieved with ureteroscopic removal of mid and proximal ureteral calculi. Thus, ureteroscopic treatment should be considered a treatment of choice for upper ureteral calculi. PMID- 7490893 TI - Phosphotyrosine antibodies preferentially react with basal epithelial cells in the dog prostate. AB - PURPOSE: Considering the hypothesis that androgen-independent but growth factor dependent epithelial cell division may be important in the development and progression of prostate cancer and that protein tyrosine kinases and phosphotyrosine protein phosphatases are key enzymes modulating the levels of specific phosphotyrosylated proteins implicated in several growth factor regulated signal transduction pathways, our aim was to study the cellular distribution of phosphotyrosine proteins in normal and hyperplastic dog prostates as well as in those of castrated dogs supplemented with either androgens or estrogens in order to modify the relative proportion of basal versus secretory epithelial cells. MATERIALS AND METHODS: Following the determination of optimal conditions to specifically detect phosphotyrosine proteins by a rabbit polyclonal antibody directed against phosphotyrosine, immunohistochemistry was performed on prostate tissue sections from these experimental animals. In addition to morphological criteria, an antibody to high molecular weight cytokeratins and antisera against arginine esterase were used to selectively identify basal and secretory cells. Since prostatic acid phosphatase may be involved in the local regulation of phosphotyrosine proteins, its distribution was also evaluated with a human prostatic acid phosphatase antiserum. RESULTS: In all prostatic tissues examined, basal epithelial cells were preferentially and specifically stained with antiphosphotyrosine. The staining intensity per basal cell was highest in the estrogen-supplemented dogs. In addition, basal cells were numerically increased and all were highly immunoreactive for high molecular weight cytokeratins. In prostates displaying a well-differentiated glandular epithelium, the number of positive basal cells and their staining intensity varied in the following order: normal < hyperplastic < androgen-supplemented dogs. At all times, the levels of phosphotyrosine proteins in prostatic acid phosphatase and arginine esterase positive cells (secretory) remained low. Fibroblasts and smooth muscle cells were unreactive to antiphosphotyrosine, even though estrogen supplementation increased the prostatic stromal volume. CONCLUSIONS: The preferential localization of phosphotyrosine proteins in basal cells, their increased level per cell and the number of positive cells in the different experimental animals support the concept that basal cells represent the stem cells of the prostate. The sex steroid-mediated up-regulation of protein phosphorylation on tyrosine residues in these cells suggests that their proliferation is likely to involve growth factor regulated signal transduction pathways. In this respect, the lack of maturation of basal cells and the differentiation of secretory cells induced by androgen deprivation, combined with estrogen stimulation, favors the activation of these pathways and cell growth. On the other hand, the activation of glandular cell differentiation and the increase of stromal volume do not alter the threshold level of protein tyrosine kinase and phosphatase activities in secretory cells, fibroblasts and smooth muscle cells. PMID- 7490894 TI - Patient controlled analgesia for shock wave lithotripsy: the effect of self administered alfentanil on pain intensity and drug requirement. AB - PURPOSE: Second generation lithotriptors offer immersion-free treatment and a decrease in shock wave induced pain. Pain sensations caused by advanced lithotriptors vary widely and have a significant impact on clinical management. We tested patient controlled analgesia during extracorporeal shock wave lithotripsy (ESWL) and quantified analgesic requirements by means of patient controlled analgesia during ESWL of renal stones. MATERIALS AND METHODS: Patients with renal stone disease undergoing ESWL were randomized prospectively to receive an alfentanil infusion titrated by 4 different anesthesiologists not otherwise involved in the study (22 controls) or to self-administer alfentanil via a patient controlled infusion pump (22 patients). As a measure of individual pain sensitivity the detection, pain and tolerance thresholds of electrocutaneous sensitivity were determined in all patients. RESULTS: Alfentanil was used more often in the patient controlled analgesia group than in the control patients (12 versus 8 required the narcotic, respectively, p = 0.226). Patients using patient controlled analgesia needed less drug (0.5 versus 2.15 mg., p = 0.005, median values), tolerated higher discharge voltages and pain intensities, needed significantly fewer shock waves to complete stone fragmentation (1,612 versus 2,105, p = 0.014) and had shorter treatment times (36.9 versus 46.2 minutes, p = 0.069). There was a significant correlation between the duration of shock wave exposure tolerated without analgesia, and pain (p = 0.0009) and tolerance (p = 0.0020) thresholds but not with regard to detection thresholds (p = 0.1400). Male patients showed significantly higher tolerance thresholds to electrocutaneous stimulation (10.2 versus 6.9 mA., p = 0.0238), which corresponded to longer analgesia-free treatment times in male versus female patients (31.7 versus 19.4 minutes, p = 0.0510). CONCLUSIONS: Patient controlled analgesia increases pain tolerance, decreases narcotic requirements, simplifies ESWL as an outpatient procedure and can be used to quantify analgesic requirements during lithotripsy. Pain and tolerance thresholds of electrocutaneous sensitivity are sensitive markers of pain tolerance during lithotripsy, which may be more pronounced in male patients. PMID- 7490895 TI - Effects of extracorporeal shock wave lithotripsy on plasma concentrations of endothelin and renin in humans. AB - PURPOSE: There is ongoing controversy regarding blood pressure changes after extracorporeal shock wave lithotripsy (ESWL*). Experimental data suggest a role for renin but only few data are relevant to humans. It has been shown that renin secretion is stimulated by endothelin, a recently discovered peptide with strong vasoconstrictive properties and stimulating effects on renin secretion. Endothelin is relevant in the development of hypertension and acute renal failure. MATERIALS AND METHODS: In a prospective study of 48 normotensive patients undergoing ESWL for renal stones the influence of high energy shock waves on plasma endothelin and active renin was analyzed. These substances are secreted by renal cells in response to hemodynamic alterations, and inflammatory and traumatic processes. Peripheral blood samples were analyzed for active renin and endothelin before, and immediately, 1, 3 and 5 days after ESWL. Blood pressure was measured before, and 1, 3 and 5 days after ESWL. RESULTS: Only a slight and transient increase was noted in active renin, which was in the same range as that found after mental stress. Endothelin and blood pressure were not significantly influenced by ESWL. There was no correlation between endothelin and active renin. Thus, the increase in active renin was not mediated by endothelin. CONCLUSIONS: The transient increase in active renin cannot be attributed to the development of hypertension. The lack of influence of ESWL on endothelin indicates that ESWL, at least in the routine clinical setting, does not cause severe renal trauma. PMID- 7490896 TI - Minimally invasive treatment of renal abscess. AB - PURPOSE: We critically evaluated the most appropriate management of renal abscesses, and identified the set of patients that most benefits from conservative treatment. MATERIALS AND METHODS: We retrospectively reviewed charts regarding discharge diagnoses, radiological studies, pathological specimens, epidemiology factors and outcomes. Statistical analysis was performed using loglinear and covariant analysis. RESULTS: Nine years of experience (1984 to 1993) at 2 affiliated hospitals (1 public and 1 private) were reviewed. A total of 52 patients with renal abscesses was identified with a followup rate of 98%. In immunocompetent patients 100% of small abscesses (less than 3 cm.) managed by antibiotics and observation alone resolved. Of medium abscesses (3 to 5 cm.) treated with percutaneous abscess drainage alone 92% resolved. Large abscesses (greater than 5 cm.) often required more than 1 percutaneous drainage procedure (33%) or adjunct open surgical intervention (37%). Statistical analysis revealed that no single treatment modality yielded a superior resolution rate or shorter hospitalization for abscesses stratified by size, patient age or treatment instituted early (1984 to 1993) or late (1992 and 1993) in the study period. CONCLUSIONS: Our series suggests that percutaneous drainage is as effective as open surgery for large and medium renal abscesses. Small abscesses may be effectively treated with a course of intravenous antibiotic therapy. A treatment algorithm is reported. PMID- 7490898 TI - Proximal ureteral stent migration: an avoidable complication? AB - PURPOSE: The parameters responsible for proximal ureteral stent migration were quantitated. MATERIALS AND METHODS: We compared 16 patients seen between 1990 and 1993 with stent migration to 32 patients whose stents did not migrate. RESULTS: The stent indwelling time and number of stents with a proximal curl of less than 180 degrees were similar between the groups. More patients with migration had stents that were shorter than the ideal length, with a distal curl of less than 180 degrees and a proximal curl in the upper calix versus the renal pelvis. CONCLUSIONS: A shorter than ideal stent, inadequate distal curl and a proximal curl in the upper calix appear to be significant factors in the process of stent migration. PMID- 7490897 TI - Endourology--1996. PMID- 7490899 TI - Long-term incidence and risks for recurrent stones following contemporary management of upper tract calculi in patients with a urinary diversion. AB - PURPOSE: The long-term rate of stone recurrence following contemporary minimally invasive management of calculi was determined in patients with urinary diversion and to identify specific risk factors for these recurrences. MATERIALS AND METHODS: We followed for a minimum of 12 months 25 patients (29 renal units) with upper tract calculi associated with urinary diversion, and treated with extracorporeal shock wave lithotripsy and/or percutaneous nephrostolithotomy. The risk of recurrent stones at 5 years was determined by a Kaplan-Meier estimate, while potential risk factors for recurrence were analyzed using a log-rank comparison. RESULTS: Stones recurred in 8 patients (32%) 18 to 61 months (mean 27.0) after treatment. The risk of new stone formation after 5 years was estimated to be 63.3% and was significantly greater in patients with recurrent bacteriuria after treatment. Neither stone history, type of procedure used, stone composition nor radiographic status at completion of treatment significantly influenced this risk. CONCLUSIONS: Contemporary management of upper tract calculi in patients with a urinary diversion is associated with a high recurrence rate, especially among those with recurrent bacteriuria, regardless of whether the patient is initially rendered stone-free. Continued close surveillance and antibiotic prophylaxis seem to be essential in this high risk patient population. PMID- 7490900 TI - Urinary calculi as a late complication of the Indiana continent urinary diversion: comparison with the Kock pouch procedure. AB - PURPOSE: Although urinary calculi have been frequent late complications of the Kock continent urinary diversion, they have not been regarded as significant problems in patients with the Indiana pouch because of the lack of foreign material present. However, stones developed in a significant percentage of our patients with an Indiana pouch. We investigated the characteristics of stone formation in patients with the Indiana pouch and compared them to those with a Kock pouch. MATERIALS AND METHODS: Detailed clinical courses regarding stone formation were reviewed in 72 patients with a Kock pouch and 54 with an Indiana pouch who had a minimum followup of 12 months. RESULTS: Stones developed in 7 of 54 patients (12.9%) with an Indiana pouch compared to 31 of 72 (43.1%) with a Kock pouch. The incidence gradually increased with longer followup but it was lower in the Indiana than in the Kock pouch group (5-year stone-free rate 84% versus 66%, respectively). Although the stones consisted principally of a mixture of struvite, carbonate apatite and ammonium hydrogen urate, variable amounts of calcium oxalate were identified in 50% of the Indiana pouch calculi. CONCLUSIONS: Not only urinary infections but also metabolic factors were considered to be involved in stone formation within the Indiana pouch. However, the substantially higher rate of stone formation in our Kock and Indiana pouch groups than has been reported in the United States suggested that no or infrequent pouch irrigations in our patients were important risk factors for urinary calculi. PMID- 7490901 TI - Nephrolithiasis in patients with urinary diversion. PMID- 7490902 TI - No evidence of osteopenia 5 to 8 years after ileal orthotopic bladder substitution. AB - PURPOSE: The use of bowel segments as bladder substitutes may result in chronic, impaired vitamin D and calcium metabolism, and ultimately in bone demineralization. MATERIALS AND METHODS: Bone metabolism was examined in 14 patients who lived for 5 to 8 years with an ileal low pressure bladder substitute after radical cystectomy for bladder cancer. Bone mineral density was measured using dual energy x-ray absorptiometry of the total skeleton, lumbar spine, femoral neck, and tibial epiphysis and diaphysis. Laboratory studies included serum levels of 1,25-dihydroxyvitamin D, 25-hydroxyvitamin D, intact parathyroid hormone, plasma alkaline phosphatase, electrolytes, creatinine and blood gas analysis. RESULTS: Bone mineral density was normal in all patients. There was no evidence of deficient vitamin D stores. There was a tendency toward slightly elevated serum creatinine values in patients with preexisting impaired renal function, including 1 who also had slight acidosis. No patient had hyperchloremia. CONCLUSIONS: We found no evidence of osteomalacia, osteoporosis or significant metabolic acidosis in 14 patients with an ileal bladder substitute for 5 to 8 years. However, it is not known whether the absence of osteopenia would also apply to patients with poor renal function, to those not followed meticulously and, thus, at risk for major long-term functional or metabolic disturbances from the ileal bladder substitute or to patients with orthotopic bladder substitutes made from longer or other bowel segments than we used. PMID- 7490903 TI - The ileal neobladder in women: 9 years of experience with 18 patients. AB - PURPOSE: We present our surgical and functional experience with orthotopic bladder replacement in women. MATERIALS AND METHODS: Since 1986, 18 women have undergone lower urinary tract reconstruction with an ileal neobladder. A nerve sparing cystectomy is done, and reservoirs are connected to the proximal urethra or urethrovesical junction. A total of 13 patients was available for complete followup as of March 1995. RESULTS: There were no perioperative deaths and few early complications. The only 2 failures were a neobladder vaginal fistula and these cases, which were converted to a conduit, are excluded from this study. Late complications requiring rehospitalization or reoperation in 2 patients included urethroileal stenosis that had to be dilated without further sequelae and bilateral ureteroileal stenosis that was treated endoscopically. At 3 months postoperatively excellent continence was achieved in 8 patients, while 2 had grade 1 stress incontinence and 3 were hypercontinent. As of March 1995 only 4 patients voided to completion while 9 required intermittent catheterization (continuously in 5 and twice daily for residual urine in 4). We were unable to demonstrate a functional difference of the various resection lines located at the proximal urethra or urethrovesical junction. CONCLUSIONS: Urethral support and nerve sparing cystectomy plus the ileal neobladder as a reservoir guarantee excellent continence in all patients. Despite our efforts, we have been unable to demonstrate any advantage of the nerve and urethral support sparing cystectomy technique as far as micturition is concerned. The development of hypercontinence in 70% of the patients with time demonstrates that our current understanding of the functional and anatomical basics of the voiding process is too limited to allow bladder replacement with a perfect functional result in all female patients. Our long-term experience, which is different from initial reports, justifies creation of an ileal neobladder in select female patients as long as they accept a 70% risk of clean intermittent catheterization in the long term. Overall patient satisfaction, including sexual life, is exceptional. However, disappointment is considerable when clean intermittent catheterization is required after periods of successful voiding per urethram. PMID- 7490905 TI - Bladder detrusor endometriosis: clinical and pathogenetic implications. AB - PURPOSE: We examined the pathogenesis of vesical endometriosis, identified the diagnostic signs and defined a successful management strategy. MATERIALS AND METHODS: The records of 8 patients with bladder detrusor endometriosis were reviewed. RESULTS: Two distinct forms of the condition appear to exist, that is spontaneous and post-cesarean. In the former case the bladder lesion is a manifestation of a generalized pelvic disease, whereas after iatrogenic dissemination growth of ectopic endometrium is usually limited to the bladder wall. The catamenial nature of bladder symptoms (frequency, urgency, dysuria and tenesmus) was pathognomonic. Cystoscopy with biopsy was diagnostic in 3 cases. Ultrasonography revealed an endo-luminal vegetation and ruled out an anterior uterine leiomyoma, whereas magnetic resonance imaging did not add relevant information. Partial cystectomy appears to cure the urinary disturbances. CONCLUSIONS: We suggest a high index of suspicion of vesical endometriosis in all premenopausal women complaining of catamenial bladder symptoms with negative urine cultures. PMID- 7490904 TI - Estimation of bladder volume using portable ultrasound in clam enterocystoplasty patients. AB - PURPOSE: We evaluated portable ultrasound for the measurement of residual volume following clam enterocystoplasty. MATERIALS AND METHODS: Bladder volumes were measured in 25 clam enterocystoplasty patients using a portable ultrasound machine and compared with urethral catheter volumes. A similar study was performed on a control group of 50 patients with normal bladder architecture. RESULTS: There was a close correlation (r = 0.978) between ultrasound and catheter volumes in the control group with a regression line slope of 0.88. There was an equally close correlation (r = 0.960) in the clam enterocystoplasty group but the regression line slope was lower at 0.65, which suggests an underestimation of the true bladder volume. There was a significant difference (p < 0.0001) between the ultrasound-to-catheter volume ratio in the 2 groups. CONCLUSIONS: Portable ultrasound is inaccurate for the measurement of residual bladder volume in clam enterocystoplasty patients. PMID- 7490906 TI - Discrimination between the ulcerous and the nonulcerous forms of interstitial cystitis by noninvasive findings. AB - PURPOSE: We determined whether the classification of interstitial cystitis cases into ulcer and nonulcer categories from cystoscopic findings could be corroborated with epidemiological data relating to demographics, risk factors, symptoms, pain and psychosocial factors. MATERIALS AND METHODS: We surveyed 565 interstitial cystitis patients (111 with and 454 without ulcer), and from univariate analyses we found 10 noninvasive variables to be significantly associated with the presence of Hunner's ulcers. The 3 multivariate statistical methodologies of discriminant analysis, logistic regression and recursive partitioning were used to classify ulcer versus nonulcer cases from these variables. The retrospective and prospective performances from each methodology were also analyzed. RESULTS: Retrospectively, resubstitution error rates of all 3 methodologies were small, with overall misclassification rates of 19.1% with linear discriminant, 14.7% with logistic regression and 8.0% with recursive partitioning. Prospectively, the overall misclassification rates increased slightly to 20.0% with linear discriminant, 15.8% with logistic regression, and 15.9% with recursive partitioning. CONCLUSIONS: The classification of interstitial cystitis into distinct categories of Hunner's ulcer (classic interstitial cystitis) and nonulcer based on cystoscopic findings can be corroborated with epidemiological evidence. The 2 categories may represent different manifestations of the underlying disease pathophysiology. PMID- 7490907 TI - Up-front intravesical chemotherapy for low stage, low grade recurrent bladder cancer. AB - PURPOSE: We evaluated the ablative and prophylactic potential of short schedule, up-front topical chemotherapy on low stage and grade recurrent bladder tumors. MATERIALS AND METHODS: The study design consisted of 4 weekly instillations followed by transurethral resection during week 5. Mitomycin C was administered initially and mitoxantrone was administered following the same schedule if disease recurred during followup. RESULTS: After up-front mitomycin C, 29 of 42 patients (69%) had a complete response with no need for transurethral resection, whereas residual disease was resected in the remaining 13 (31%). Disease recurred during followup in 22 of the 42 patients (52.4%), who were then treated with up front mitoxantrone with a complete response in 14 (63.7%). Residual disease was resected in 8 patients (36.3%) with progression to grade 3 in 2. CONCLUSIONS: Short schedule intravesical chemotherapy can completely ablate small volume recurrent superficial bladder cancer in a relevant number of patients but it is not adequate prophylaxis. PMID- 7490908 TI - Histological grading of carcinoma in situ of the bladder: its clinical significance in patients who underwent intravesical mitomycin C and doxorubicin sequential therapy. AB - PURPOSE: The clinical behavior of carcinoma in situ of the bladder seems rather complicated. Although some have advocated the histological grading of carcinoma in situ, to our knowledge no sufficient clinical information has been reported. Therefore, we evaluated the clinical significance of histological grading of carcinoma in situ of the bladder. MATERIALS AND METHODS: From January 1984 to December 1991, 58 patients with carcinoma in situ of the bladder were treated initially with intravesical mitomycin C and doxorubicin sequential therapy. Of the patients 20 had grade 2 and 38 had grade 3 anaplasia according to the modified World Health Organization grading system. Those who failed the initial therapy received another course of mitomycin C and doxorubicin sequential therapy or intravesical bacillus Calmette-Guerin. RESULTS: Following initial therapy, 13 patients (65%) with grade 2 and 28 (74%) with grade 3 disease achieved a complete response. Subsequent intravesical therapy resulted in complete response in 17 patients (85%) with grade 2 and 31 (82%) with grade 3 cancer. The local recurrence rate was higher in the grade 2 than in the grade 3 cases after a median followup of 48 months (range 10 to 84). The recurrent tumor configuration was significantly different between the 2 groups. Papillary cancer recurred only in grade 2 cases, while only nodular cancer recurred in grade 3 cases. The progression-free and survival curves were slightly higher in grade 2 than in grade 3 cases, although the difference was not significant. CONCLUSIONS: There may be some difference in response to initial intravesical chemotherapy and the local recurrence rate between grades 2 and 3 carcinoma in situ, both of which are detrimental to grade 2 lesions. Moreover, it appears likely that grade 2 carcinoma in situ is a precursor of papillary high grade cancer and grade 3 carcinoma in situ is a precursor of nodular cancer. However, patient prognosis in the 2 groups was not significantly different. PMID- 7490910 TI - Inflammatory pseudotumor of the lacrimal sac. AB - A case of inflammatory pseudotumor of the lacrimal sac is presented. The patient was first consulted for epiphora and swelling of the right lacrimal sac region. A mass was found on computed tomography (CT) and finally it was removed by dacryocystectomy. The diagnosis of inflammatory pseudotumor was confirmed by histopathological sections. PMID- 7490911 TI - delta-Aminolevulinic acid dehydratase deficiency porphyria (ADP) with syndrome of inappropriate secretion of antidiuretic hormone (SIADH) in a 69-year-old woman. AB - delta-Aminolevulinic acid dehydratase deficiency porphyria (ALAD porphyria, ADP) with syndrome of inappropriate secretion of antidiuretic hormone (SIADH) in a 69 year-old woman is reported. The patient was admitted to our hospital complaining of slight cough with low-grade fever, and treated with piperacillin sodium, resulting in complete resolution of the symptoms, following a diagnosis of bronchopneumonia. Thereafter, however, she began to complain of vomiting, abdominal pain, facial numbness and paresis of the extremities with gait disturbance, and became comatose with hyponatremia (serum Na concentration 119 mEq/L) in a few days. Laboratory tests revealed an antidiuretic hormone (ADH) level of 13.5 pg/mL, plasma osmolality 218 mOsm/KgH2O, urinary osmolality 429 mOsm/KgH20, urinary Na concentration > 20 mEq/L, and no abnormalities of thyroid, adrenal or renal function. Neither edema nor dehydration was evident. These data indicated the presence of SIADH. No abnormalities suggestive of malignant or infectious diseases such as lung cancer, pneumonia and Guillain-Barre syndrome were evident from laboratory and roentgenographic findings. As the cause of SIADH, therefore, porphyria was suspected. Metabolites and activities of enzymes in the heme biosynthetic pathway were examined, and very low activity of delta aminolevulinic acid dehydratase (ALA-D) (0.14 mumol PBG/mL RBC/h) was found. The patient was neither an alcoholic nor a heavy smoker, and she had no past history of heavy metal intoxication, photosensitivity or tyrosinemia. On the basis of these data and clinical features, she was diagnosed as having ADP. We consider this to be the first case of ADP reported in Japan. PMID- 7490909 TI - The effect of iron deficiency and mental stimulation on Indonesian children's cognitive performance and development. AB - Three studies conducted in Indonesia will be described. Soewondo (12) investigated the relation of iron deficiency and cognitive function and impact of iron supplementation on verbal intelligence, attention and concept learning among iron deficient children without anemia and iron deficient anemic children. Half of 176 children, aged 3-6 years, received elemental Fe for 8 weeks and the other half received placebo. There were significant changes from pre to post intervention evaluations in ferritin, transferrin saturation, free erythrocyte protoporphyrin, and hemoglobin in the iron deficient anemic children. Pre and post treatment psychological test data showed that iron deficiency anemia produced alterations in cognitive processes related to visual attention and concept acquisition. These alterations can be reversed with iron treatment. Idjradinata (4) assessed the impact of iron supplementation on iron deficient infant's mental and psychomotor development. Hundred twenty six subjects aged 12 to 18 month were randomly assigned to either iron treatment or placebo intervention. After 4 months of iron supplementation, the hemoglobin, ferritin and transferrin saturation changed significantly in the iron deficient infants. A developmental delay was observed in the iron deficient anemic infants before intervention and the conditions were reversed after 4 months of iron treatment. Soemiarti (8) examined the effectiveness of a training course given to mothers of children aged 12 to 24 month on the rearing environment and consequently to the child's development. The subjects were 69 mothers of 20-35 years old. The training lasted for 21 days by giving mothers training using the program "Ibu Maju Anak Bermutu". The rearing environment improved, also the child's mental and psychomotor development. PMID- 7490912 TI - Karl Theodore Dussik--pioneer in ultrasound. PMID- 7490913 TI - Accuracy of frozen section diagnosis in surgical pathology: review of a 1-year experience with 24,880 cases at Mayo Clinic Rochester. AB - OBJECTIVE: To determine the accuracy of frozen section examination for routine diagnostic use in surgical pathology. DESIGN: We retrospectively reviewed the experience with frozen sections at Mayo Clinic Rochester during calendar year 1993. MATERIAL AND METHODS: The results in 24,880 cases (97,914 frozen section slides) processed during 1993 were compared with findings on permanent sections, and types of errors detected were classified into one of three quality assurance categories. RESULTS: The overall rate of frozen section accuracy was 97.8%. Of the 2.2% of surgical pathology reports that needed revision, 1.6% were unavoidable tissue sampling errors, 0.5% were errors that reflected change in degree of abnormality, and only 0.1% were clinically significant errors that may have affected patient management. CONCLUSION: Frozen section diagnosis is accurate for processing a high volume of surgical pathology cases. PMID- 7490914 TI - General surgical laparoscopic procedures for the "nonlaparologist". AB - OBJECTIVE: To assess our initial experience with a variety of laparoscopic procedures that can be performed by general surgeons with the basic skills used for laparoscopic cholecystectomy and without advanced laparoscopic expertise. MATERIAL AND METHODS: We retrospectively reviewed a 36-month experience (1991 through 1994) at our institution with a consecutive series of patients who underwent basic laparoscopic surgical procedures but specifically excluding cholecystectomy, appendectomy, herniorrhaphy, and colectomy. RESULTS: Procedures performed laparoscopically included gastrostomy, jejunostomy, small bowel resection, intra-abdominal and retroperitoneal biopsy, staging of intra-abdominal malignant lesions, and adhesiolysis for relief of small bowel obstruction. During the 3-year study period, 106 patients underwent 107 procedures, 89 of which were successful. Four patients had substantial complications, two of whom underwent surgical repair. CONCLUSION: A spectrum of procedures may be safely performed with the skills learned from laparoscopic cholecystectomy and without the need for advanced laparoscopic skills such as intracorporeal suturing or tying of knots. PMID- 7490915 TI - DNA fluorescent probes for diagnosis of velocardiofacial and related syndromes. AB - OBJECTIVE: To study the usefulness of fluorescent in situ hybridization (FISH) with the DNA probe D22S75 for detecting microdeletions in chromosome 22q11.2 in metaphases from patients with features of "CATCH 22" (cardiac anomalies, abnormal facies, thymic hypoplasia or aplasia, cleft palate, and hypocalcemia). METHODS: High-resolution chromosome analysis and FISH were performed on metaphases from 10 control subjects, 42 patients with features of CATCH 22, and 6 parents of children with CATCH 22. Patients were screened for conotruncal heart defect, palatal abnormality, and facial features. We correlated the phenotype, karyotype, and deletion of a D22S75 locus. RESULTS: Specimens from nine patients with one or more features of CATCH 22 had a single hybridization signal for D22S75, indicating a deletion of chromosome 22q11.2. Four patients had all the major features of the syndrome and a chromosomal deletion. Thirteen patients had two CATCH 22 features, five of whom had a deletion. None of the 25 patients with a single CATCH 22 feature had a deletion. One patient with a deletion detected by FISH also had a deletion noted on high-resolution banding. All six parents who had blood samples studied by FISH had normal hybridization patterns. CONCLUSION: FISH is a useful adjunct to chromosome analysis for assessing patients with features of CATCH 22. Detecting a chromosomal deletion by FISH provides a definitive diagnosis and helps to ensure appropriate medical management and genetic counseling. PMID- 7490916 TI - HLA-DQ associations in type 1 autoimmune hepatitis. AB - OBJECTIVE: To determine whether, in patients with type 1 autoimmune hepatitis and human leukocyte antigen (HLA) DR3 and DR4 positivity, any DQ antigen is disease specific. MATERIAL AND METHODS: HLA class II typing was performed by restriction fragment length polymorphism in 103 patients with type 1 autoimmune hepatitis, 104 patients with chronic viral hepatitis, and 80 normal subjects. A shared association with a disease-specific DQ antigen was sought in patients with HLA DR3, DR4, and DR3-DR4. RESULTS: Patients with HLA-DR3 and DR4 shared positivity for DQ2, DQ4, DQ5, DQ6, and DQ7, but the associations reflected established linkages or were of low frequency. Patients heterozygous for DR3-DR4 or homozygous for either DR3 or DR4 did not have a shared DQ antigen. Only the DR3 DQ2 haplotype distinguished patients with autoimmune hepatitis from normal subjects or those with chronic viral hepatitis. CONCLUSION: The DR3 and DR4 antigens are not associated with a single disease-specific DQ antigen in type 1 autoimmune hepatitis. The DR3-DQ2 haplotype is the principal risk factor for the disease at our referral center. Analyses by restriction fragment length polymorphism do not implicate a single susceptibility gene at the DQ locus. PMID- 7490917 TI - Extramedullary hematopoiesis manifesting as a symptomatic pleural effusion. AB - The occurrence of extramedullary hematopoiesis in the pleura is rare and is usually asymptomatic. It is generally diagnosed on postmortem examination. Herein we describe a 61-year-old woman with agnogenic myeloid metaplasia who sought medical assessment because of progressive dyspnea. Thoracentesis yielded sanguineous fluid with 10% myeloblasts. A 99mTc bone marrow scan demonstrated increased tracer activity throughout both lungs, an outcome consistent with extramedullary hematopoiesis. Fine-needle aspiration and video-assisted thoracoscopy were considered but deferred because of the potential risk of profuse bleeding. Treatment with low-dose external beam irradiation to the left posterior pleura was successful. To date, no hemothorax has recurred. PMID- 7490918 TI - Maximizing vocational outcome after brain injury: integration of medical and vocational hospital-based services. AB - OBJECTIVE: To describe a medical and vocational case-management system that has been implemented at the Mayo Medical Center in Rochester, Minnesota, to decrease the time between onset of brain injury and successful community reintegration. DESIGN: Barriers to employment are identified and targeted as part of a system that integrates medical center- and community-based services. MATERIAL AND METHODS: The success of the project will be determined by several sources, including the Mayo-Portland Adaptability Inventory, a vocational rating scale, and traditional measures of vocational success (for example, level of independent living, job type and setting, rate of pay, and the type and cost of vocational supports being used). Overall study outcomes will be compared against benchmarks derived from the literature on vocational outcome after brain injury. RESULTS: Preliminary data are encouraging that project goals are attainable. The medical case-management system has decreased the amount of time between injury and initiation of vocational and other rehabilitation services. During the first year, 67 persons with brain injury began receiving vocational services through the project, and 34% are in community-based nonsheltered work or training programs. Second-year data will reveal whether the goal of 70% placement within 9 months after admission to the project can be maintained. PMID- 7490919 TI - Current concepts in airway management for cardiopulmonary resuscitation. AB - OBJECTIVE: To describe the methods of maintaining airway patency for oxygenation during cardiopulmonary resuscitation (CPR) that do not require expertise in mask ventilation or endotracheal intubation by direct laryngoscopy. DESIGN: A review of rescue breathing and newer methods of providing airway patency is provided. RESULTS: Airway patency during CPR is often difficult to achieve. Mask ventilation predisposes to hypoventilation and aspiration pneumonitis. Endotracheal intubation by direct laryngoscopy is the preferred method of maintaining airway patency for CPR. Alternative techniques for airway management include endotracheal intubation by lighted stylet, esophageal tracheal Combitube, laryngeal mask airway, and transtracheal ventilation. These methods are recommended by the American Heart Association and the American Society of Anesthesiologists. They have been approved by the Food and Drug Administration for maintenance of airway patency; they are easy to learn, effective, and applicable to CPR. Advantages and disadvantages of each technique may indicate or contraindicate one method over another in specific circumstances. CONCLUSION: When CPR is compromised by airway obstruction that remains unresponsive to traditional techniques, using alternative methods is appropriate. The techniques selected must be based on individual familiarity and expertise. PMID- 7490920 TI - Acute myocardial infarction: enhancing the results of reperfusion therapy. AB - Reperfusion therapy (by either administration of thrombolytic agents or immediate angioplasty) is the mainstay of treatment of acute myocardial infarction, yet it remains underutilized. Additionally, many patients do not experience optimal reperfusion because of suboptimal flow rate in the infarct-related artery, abnormal microvascular flow, and intermittent or complete reocclusion of the infarct-related artery. Strategies to enhance the results of reperfusion therapy include expanding the population of patients considered as candidates, earlier treatment, newer methods to improve infarct-related artery flow rates, and ancillary treatments currently being studied to reduce reperfusion injury, which may be one cause of decreased microvascular flow. Although aspirin, heparin, and warfarin sodium have been the conventionally used agents for inhibiting thrombin and platelet function, newer agents such as hirudin and inhibitors of the platelet glycoprotein IIb-IIIa receptor are becoming available, and their clinical application will increase in the future. PMID- 7490921 TI - 39-year-old man with fever, cough, and chest pain. PMID- 7490922 TI - Genetic testing for Prader-Willi and Angelman syndromes. PMID- 7490923 TI - Rehabilitation of geriatric patients. AB - OBJECTIVE: To describe the role of geriatric rehabilitation, the methods of practice, and the mechanisms underlying integrated care in rehabilitation medicine. DESIGN: We reviewed the rehabilitation-related medical literature and outlined an approach to the evaluation of impairment in older patients and subsequent rehabilitation planning. RESULTS: For optimal rehabilitation of geriatric patients, knowledge of general principles of rehabilitation and key areas affecting the elderly population is important. A rehabilitation program will be most successful if a team approach--including a physician, rehabilitation nurse, physical therapist, occupational therapist, psychologist, medical social worker, and recreational therapist--is used. Attention should be paid to special considerations in older patients (such as the presence of comorbid conditions) and potential barriers to care in order to maximize benefit. CONCLUSION: Rehabilitation of elderly patients can assist in preserving functional independence and improving the quality of life. Assessment and management in an individual patient are best performed by a team approach. PMID- 7490924 TI - Common rheumatologic diseases in elderly patients. AB - OBJECTIVE: To review common rheumatologic disorders that affect elderly patients and emphasize the unique diagnostic and therapeutic challenges inherent in the management of rheumatologic diseases in this age-group. DESIGN: We summarize our approach to treatment and management of specific rheumatologic problems in geriatric patients and discuss pertinent studies from the literature. RESULTS: Among the spectrum of rheumatologic disorders frequently encountered in the elderly population are polymyalgia rheumatica, fibromyalgia, giant cell arteritis, crystalline arthropathies (gout and pseudogout), and degenerative joint disease. The initial manifestations of these rheumatologic diseases in elderly patients may differ from the typical findings in younger patients. Geriatric patients may have nonspecific complaints, a decline in physical function, or even confusion. Because of physiologic changes associated with aging and a decrease in functional reserves, elderly patients are susceptible to adverse effects of pharmacologic therapy (including nonsteroidal anti inflammatory medications, corticosteroids, narcotic analgesics, allopurinol, and colchicine). Clinicians should be alert for such problems as hepatotoxicity and occult gastrointestinal blood loss. Comorbid conditions such as cardiovascular disease and cognitive impairment may complicate management strategies and may limit the goals of both surgical intervention and rehabilitation programs in elderly patients. CONCLUSION: Rheumatologic disorders in geriatric patients pose special challenges to primary-care physicians. In the selection of optimal pharmacologic and nonpharmacologic therapeutic modalities, clinicians should focus on maintaining or improving the patient's quality of life and level of independent function. PMID- 7490925 TI - A tribute to Russell Daniel Carman. PMID- 7490926 TI - Genitourinary tract radiology at the Mayo Clinic: history portrayed through medical literature. PMID- 7490927 TI - Neuroradiology at the Mayo Clinic. PMID- 7490929 TI - Management of alcohol withdrawal. PMID- 7490928 TI - Intraoperative consultations by pathologists at the Mayo Clinic: a unique experience. PMID- 7490930 TI - Management of alcohol withdrawal. PMID- 7490931 TI - Renal lymphoma and human immunodeficiency virus infection. PMID- 7490932 TI - Causes of rhabdomyolysis. PMID- 7490933 TI - [The epidemiology of the acute adverse drug reaction in Barcelona in 1983-1992: a mortality analysis]. AB - BACKGROUND: Mortality caused by acute adverse drug reactions (AADR) increased in Spain over the eighties and has become one of the major causes of death for youth. This paper presents the results of a study in the city of Barcelona during the 1983-92 decade; based in the city drug information system. METHODS: The study includes all deaths caused by AADR autopsied in the Forensic Institute between 1983 and 1992. Mortality of city residents is analyzed by cohort, grouping data in two-years and adjusting Poisson regression to mortality rates by age group, birth cohort and period of study. RESULTS: During this period a sharp increase in AADR mortality is seen, as there were 19 deaths in the city in 1983 and 160 in 1992. The increase concentrates in the years 1987-89. Although previously the increasing mortality of drug users was related by some observers with the progressive deterioration of their health status, linked to their increasing age and to the cumulative effects of years of substance dependence, the mortality analysis points to a clear period effect, resulting in an increased mortality across all age groups, for cohorts born after 1960. CONCLUSIONS: These results point to changes in the illegal drug market in the city or in the patterns of abuse as most likely causes for the increase. The results also suggest that since 1989 there is a stabilization in mortality due to AADR, which deserves further study. PMID- 7490934 TI - [the early diagnosis of the vertical transmission of the human immunodeficiency virus type 1. The evaluation of diagnostic tests]. AB - BACKGROUND: The early diagnosis of vertically transmitted human immunodeficiency virus infection cannot be based on the presence of specific serum antibodies since those of the maternal IgG class pass the placenta and may be detected in children for up to 18 months. Based on this fact, the aim of this study was to evaluate other techniques for early diagnosis of the infection applicable from birth in 306 children of infected mothers. METHODS: The production of in vitro antibodies, virus culture and polymerase chain reaction (PCR) were used. The sensitivity of the techniques was estimated in the 40 children diagnosed with human immunodeficiency virus infection and specificity was determined in the 266 uninfected children. RESULTS: The sensitivity for the production of in vitro antibodies was 62.0% at 3 months and 94.7% at 6 months; 90.4% and 88.2%, respectively, for the viral culture and 92.3% and 94.1%, respectively, for the PCR. The specificity of all the cases was higher than 89.4% although varied in relation to age. CONCLUSIONS: The combination of several diagnostic techniques provides better performance for the early diagnosis of vertical transmission of the human immunodeficiency virus. Given that viral culture takes longer to provide results and is more expensive, it is less recommendable for routine use, although the form of viral replication may be useful to establish the prognosis. PMID- 7490935 TI - [The value of the parathyrin-related protein (PTH-RP) in the diagnosis of cancer associated hypercalcemia]. AB - BACKGROUND: The parathyrine related protein (PTH-RP) is very similar, both in structure and in function, to the PTH and is considered as a mediator in humoral hypercalcemia in cancer. The aim of this study was to know the clinical value of PTH-RP measurement. METHODS: Serum PTH-RP concentrations were studied in 22 healthy subjects, 13 patients with primary hyperparathyroidism, 9 patients with solid neoplasms and normocalcemia, 26 patients with solid neoplasms and hypercalcemia and 4 patients with hematologic neoplasms and hypercalcemia. The PTH-RP was quantified by a competitive radioimmunoassay technique using a specific antibody of the PTH-RP 1-40 fragment. Intact parathyrine (i-PTH) was quantified by an IRMA method using 2 polyclonal antibodies (INCSTAR). RESULTS: Fifteen (68%) of the healthy controls presented undetectable serum PTH-RP concentrations. The serum PTH-RP concentration was normal in all those patients with hyperparathyroidism. Elevated serum PTH-RP values were not found in patients with solid neoplasms and normocalcemia or in those with hematologic neoplasms and hypercalcemia. High values of PTH-RP were observed in 8 out of 9 (88%) of the patients with solid neoplasms and hypercalcemia with bone metastasis and in 7 out of 11 (63%) of the patients with bone involvement. CONCLUSIONS: Serum parathyrine related protein was found to be high in a large proportion of patients with solid neoplasms and hypercalcemia. Serum PTH-RP determination is useful in the clinical investigation of patients with hypercalcemia. Even in patients with bone metastasis, hypercalcemia may have a humoral background. PMID- 7490937 TI - [Aminoglycoside nephrotoxicity and obstructive jaundice]. PMID- 7490936 TI - [The risk factors in death from heroin]. PMID- 7490938 TI - [A cystic lymphoepithelial lesion of the parotid in HIV-1-infected patients]. AB - Cystic lymphoepithelial lesion is an infrequent disorder involving the saliva glands mainly the parotid gland; however, its frequency has increased coinciding with the epidemics of HIV-1 infection. Three cases of parotid lymphoepithelial lesion in patients with HIV-1 infection, 2 of whom were hemophiliacs and one an ex-intravenous drug addict, are presented. In all the cases the lesion was a painless slow growth right submandibular mass, with no history of inflammation. Two of the patients had a contralateral tumor of the same characteristics which was not studied. Fine needle aspiration puncture was performed in the three patients although diagnosis was obtained by pathologic study following superficial parotidectomy. Cystic lymphoepithelial lesion is an entity to be taken into account on differential diagnosis of cervical masses in patients with HIV infection. In individuals with severe immunosuppression treatment should be conservative. PMID- 7490939 TI - [Psychosocial effects in pharmacology: something more than an anecdote?]. PMID- 7490940 TI - [The gastropathy of portal hypertension]. PMID- 7490941 TI - [Syncope due to asystole in glossopharyngeal neuralgia]. PMID- 7490942 TI - [The sexual transmission of zidovudine-resistant HIV-1 strains]. PMID- 7490943 TI - [Apropos the prevention of the vertical transmission of HIV]. PMID- 7490944 TI - [B-lymphocytes in the pleural effusion of chronic lymphocytic leukemia]. PMID- 7490945 TI - [Lhermitte's sign and cancer]. PMID- 7490946 TI - [A physician in Gothenburg wanted to test something new. A private emergency call car improved quality of life]. PMID- 7490947 TI - [The National Board of Health and Welfare approves ovum donation]. PMID- 7490949 TI - [Specialized contra non-specialized care: increased cooperation is necessary for better efficiency]. PMID- 7490948 TI - [A professor from Harvard looks about for synergy instead for conflicts: promote health and human rights--two targets that complement each other]. PMID- 7490950 TI - [Who should perform laparoscopic cholecystectomy?]. PMID- 7490952 TI - [Cost savings increase the pressure on ambulance care]. PMID- 7490953 TI - [Ginseng extracts: on the importance of significance]. PMID- 7490951 TI - [Insulin injections and the factual reality]. PMID- 7490954 TI - [Where are we when it comes to lipids?]. PMID- 7490955 TI - [Does a defective endothelial function influence blood pressure? Complex relation between endothelium and hypertension]. PMID- 7490956 TI - [Difficult to talk about psychosocial problems. A video shows how easily may the physician-patient dialogue be blocked]. PMID- 7490957 TI - [Parvovirus B19 infection in different shapes. But more laboratories have refined the diagnostic methods]. PMID- 7490959 TI - [Splenic rupture in coloscopy]. PMID- 7490958 TI - [Placebo effect or not? Vegetarian food helped the patients with rheumatism]. PMID- 7490961 TI - [The new novel by Jersild may start a Swedeish debate on euthanasia]. PMID- 7490960 TI - [Neuroborreliosis--important differential diagnosis in radicular pain]. PMID- 7490962 TI - [Nutrition is not visible in medical educations]. PMID- 7490964 TI - [The specialists' competence must be worth while the bother! Today it's a paper tiger without guts]. PMID- 7490965 TI - [Soften punishment in malpractive cases! Prevent occurrence of a culture of dissatisfaction and revenge!]. PMID- 7490963 TI - [Johan Giesecke on infection protection in Sweden: "the victory over the infectious diseases must be achieved again and again!"]. PMID- 7490966 TI - [Current statistics by the SBU are not up to date]. PMID- 7490968 TI - [Alarming lack of knowledge of pulse oximetry]. PMID- 7490967 TI - [A world report on IVF: best results in Sweden]. PMID- 7490969 TI - [Increased interest of chronic leg ulcers. Recommendations from the Medical Product Agency]. PMID- 7490970 TI - [Oral contraceptives in perspective--benefit-risk evaluation]. PMID- 7490971 TI - [Oral contraceptives and thrombosis--current risk assessment. Recommendations from the Medical Product Agency]. PMID- 7490972 TI - [A case report. Sepsis in connection with levonorgestrel IUD]. PMID- 7490973 TI - [A test for APC resistance prior to prescription of oral contraceptives? Discussion about screening should be based on facts]. PMID- 7490974 TI - [Hip dislocation and urinary tract. Two trends in pediatric radiology]. PMID- 7490975 TI - [Abuse of alcohol and central nervous system stimulants. A growing problem in developing countries]. PMID- 7490976 TI - [Correct thinking but incorrect handlings. Confusing reality of emergency care physicians doesn't fit in the responsibility pattern]. PMID- 7490977 TI - [American health services surveyed in databases. A comprehensive picture improves quality of health care]. PMID- 7490978 TI - [A British conference: health care strengthens its scientific basis]. PMID- 7490979 TI - [Social rehabilitation of patients with coronary disease. A new type of cooperation improves the results]. PMID- 7490980 TI - [Acute disseminated encephalomyelitis--ADEM. A rare, but serious disease]. PMID- 7490982 TI - [Writing colleagues. A psychiatrician from Lisbon as a Nobel Prize candidate]. PMID- 7490981 TI - [Giving hope and being sincere at the same time. Difficult to inform about uncurable disease]. PMID- 7490983 TI - Health effects of sanctions on Iraq. PMID- 7490984 TI - Victory over guineaworm disease: partial or pyrrhic? PMID- 7490986 TI - Which gene for which restenosis? PMID- 7490985 TI - Evidence-based management of dyslipidaemia. PMID- 7490987 TI - Premenstrual syndrome no progesterone; premenstrual dysphoric disorder no serotonin deficiency. PMID- 7490988 TI - Upper limb disease in women treated for breast cancer. PMID- 7490990 TI - Case-control study of antenatal and intrapartum risk factors for cerebral palsy in very preterm singleton babies. AB - The increase in survival of very preterm babies during the 1980s was accompanied by a sharp increase in the rate of cerebral palsy in this group. The relation between antenatal and intrapartum factors and cerebral palsy in such babies has not been well defined. To identify adverse and protective antenatal and intrapartum factors we undertook a case-control study of 59 very preterm babies who developed cerebral palsy, identified from a population-based register, and 234 randomly selected controls. The frequency of cerebral palsy decreased with increasing gestational age and birthweight. Antenatal complications occurred in 215 (73%) of the women with preterm deliveries. Factors associated with an increased risk of cerebral palsy after adjustment for gestational age were chorioamnionitis (odds ratio 4.2 [95% CI 1.4-12.0]) prolonged rupture of membranes (2.3 [1.2-4.2]), and maternal infection (2.3 [1.2-4..5]). Pre-eclampsia was associated with a reduced risk of cerebral palsy (0.4 [0.2-0.9]), as was delivery without labour (0.3 [0.2-0.7]). There was no increased risk of cerebral palsy with intrauterine growth retardation (1.0 [0.9-1.1]). The effect of rigorous management of adverse antenatal factors on the frequency of cerebral palsy in very preterm babies should be tested in randomised controlled trials. PMID- 7490989 TI - Outcomes in 248 patients who had diagnostic evaluations for epilepsy surgery. AB - Surgery for intractable epilepsy is a widely used treatment that is not readily assessed by randomised trials. We evaluated the impact of epilepsy surgery on seizures, medication use, employment, and the quality of life in 248 adults and adolescents consecutively referred to one medical centre between 1974 and 1990. Outcomes were determined through self-administered questionnaire and medical record review for 202 surgery and 46 non-surgery patients whose treatment was usually determined by the presence or absence of an epileptogenic focus. Surgery and non-surgery patients differed at baseline only in median monthly seizure frequency (surgery lower than non-surgery). After adjustment for baseline covariates, surgery patients at follow-up had greater decline in average monthly seizure frequency (-11.9 vs - 1.5; difference -10.4, 95% CI -20.5, -0.3) and took fewer antiepileptic medications (average number 1.4 vs 2.0; difference -0.67, 95% CI -0.94, -0.40). Although quality-of-life scores were higher (p < 0.05) with surgery on 5 of 11 scales that were administered only at follow-up, there were no significant differences in employment status or prospectively assessed quality of life. Relative to a non-surgery group, patients treated surgically had better seizure control with less antiepileptic medication. The impact of epilepsy surgery on quality of life and employment needs to be assessed in larger prospective studies. PMID- 7490991 TI - Impact of a short course in pharmacotherapy for undergraduate medical students: an international randomised controlled study. AB - Irrational prescribing is a habit which is difficult to cure. However, prevention is possible and for this reason the WHO Action Programme on Essential Drugs aims to improve the teaching of pharmacotherapy to medical students. The impact of a short problem-based training course in pharmacotherapy, using a WHO manual on the principles of rational prescribing, was measured in an international multi-centre randomised controlled study of 219 undergraduate medical students in Groningen (Netherlands), Kathmandu (Nepal), Lagos (Nigeria), Newcastle (Australia), New Delhi (India), San Francisco (USA), and Yogyakarta (Japan). The manual and the course presented the students, who were about to enter the clinical phase of their studies, with a normative model for pharmacotherapeutic reasoning in which they were taught to generate a "standard" pharmacotherapeutic approach to common disorders, resulting in a set of first-choice drugs called P(ersonal)-drugs. The students were then taught how to apply this set of P-drugs to specific patient problems on the symptomatic treatment of pain, using a six-step problem-solving routine. The impact of the course was measured by tests before training, immediately after, and six months later. After the course, students from the study group performed significantly better than controls in all patient problems presented (p < 0.05). The students not only remembered how to solve old problems, but they could also apply their skills to new problems. Both retention and transfer effect were maintained at least six months after the training session in all seven medical schools. In view of the impossibility of teaching students all basic knowledge on the thousands of drugs available, this approach seems to be an efficient way of teaching rational prescribing. However, the method should be accompanied by a change in teaching methods away from the habit of transferring knowledge about the drugs towards problem-based teaching of therapeutic reasoning. PMID- 7490992 TI - Neurodegeneration and diabetes: UK nationwide study of Wolfram (DIDMOAD) syndrome. AB - Wolfram syndrome is the association of diabetes mellitus and optic atrophy, and is sometimes called DIDMOAD (diabetes insipidus, diabetes mellitus, optic atrophy, and deafness). Incomplete characterisation of this autosomal recessive syndrome has relied on case-reports, and there is confusion with mitochondrial genome disorders. We therefore undertook a UK nationwide cross-sectional case finding study to describe the natural history, complications, prevalence, and inheritance of the syndrome. We identified 45 patients with Wolfram syndrome--a prevalence of one per 770,000. Non-autoimmune, insulin-deficient diabetes mellitus presented at a median age of 6 years, followed by optic atrophy (11 years). Cranial diabetes insipidus occurred in 33 patients (73%) with sensorineural deafness (28, 62%) in the second decade; renal-tract abnormalities (26, 58%) presented in the third decade followed by neurological complications (cerebellar ataxia, myoclonus [28, 62%]) in the fourth decade. Other abnormalities included gastrointestinal dysmotility in 11 (24%), and primary gonadal atrophy in seven of ten males investigated. Median age at death (commonly central respiratory failure with brain-stem atrophy) was 30 years (range 25-49). The natural history of Wolfram syndrome suggests that most patients will eventually develop most complications of this progressive, neurodegenerative disorder. Family studies indicate autosomal recessive inheritance with a carrier frequency of one in 354, an absence of a maternal history of diabetes or deafness, and an absence of the mitochondrial tRNA Leu (3243) mutation. Juvenile onset diabetes mellitus and optic atrophy are the best available diagnostic criteria for Wolfram syndrome, the differential diagnosis of which includes other causes of neurodegeneration. PMID- 7490993 TI - Invasiveness of endometriotic cells in vitro. AB - The pathogenesis of endometriosis is not known. The currently favoured theory is that viable endometrial cells, shed from the endometrium into the pelvic cavity by retrograde menstruation, reattach and invade other tissues. We used a collagen gel invasion assay to assess invasive potential of endometriotic cells. The invasion indices of cells from peritoneal endometriotic lesions and a metastatic bladder carcinoma cell line (EJ28) were similar (2.2-15.6 vs 8.4-11.6) whereas cells from normal endometrium and non-metastatic carcinoma cells (RT112) were non invasive (indices < 1). Invasiveness of endometriotic cells might contribute to the pathogenesis of endometriosis. PMID- 7490994 TI - Transcutaneous oxygen levels in retinopathy of prematurity. AB - Retinopathy of prematurity (ROP) is a potentially blinding disease of preterm infants. 21 days of computer-recorded transcutaneous oxygen (TcPO2) data were compared in 31 infants with stage 3 or greater ROP and 38 infants with no ROP or stage 1 or 2. In a multiple logistic regression adjusted for significant perinatal factors (birthweight, gestation, and intraventricular haemorrhage), babies with stage 3 or higher ROP showed an increased variability of TcPO2 in week 1 (p < 0.01) and 2 (p = 0.012) but not week 3. Variability of TcPO2 in the first 2 weeks of life is a significant predictor of severe ROP. PMID- 7490995 TI - An HIV-1-positive child with fever and a scalp nodule. PMID- 7490997 TI - History and importance of scrofula. PMID- 7490996 TI - Sheffield risk and treatment table for cholesterol lowering for primary prevention of coronary heart disease. AB - When used for the secondary prevention of coronary heart disease, treatment with an inhibitor of hydroxymethylglutaryl-coenzyme-A reductase results in worthwhile benefit that clearly exceeds any risk in patients whose risk of coronary death is 1.5% or more per year. This evidence can be extrapolated logically to primary prevention of coronary disease provided that treatment is targeted at those with similar or higher risk. We present a table that refines previously proposed methods of risk prediction. The table identifies subjects who have the specified degree of coronary risk; shows the serum cholesterol concentration that confers that degree or risk in the individual; and identifies subjects who will not have this degree of risk, irrespective of their cholesterol concentration. It is simple enough for use in ordinary practice. The table highlights the predominant effect of age on coronary risk; a person who is free of vascular disease and younger than 52 years is unlikely to have the specified degree of risk. Even in older people (60-70 years) several risk factors are generally required to attain this degree of risk. Some people are candidates for lipid- lowering drug treatment with serum cholesterol as low as 5.5 mmol/L, whereas others with cholesterol as high as 9.0 mmol/L are not. Although cholesterol lowering is a powerful method for preventing coronary events in people at high risk, cholesterol measurement by itself is not a good way to identify those with high coronary risk. The method can be adapted readily to target a different level of coronary risk as new evidence on the benefit and risk of treatment becomes available. PMID- 7490999 TI - When the government shuts down. PMID- 7490998 TI - Little arrows. PMID- 7491000 TI - Drug prevents SIV infection. PMID- 7491001 TI - Piercing of dura during spinal anaesthesia. PMID- 7491003 TI - Age versus ageing as a cause of dementia. PMID- 7491002 TI - Health of Baghdad's children. PMID- 7491004 TI - Age versus ageing as a cause of dementia. PMID- 7491005 TI - Low-molecular-weight heparin for venous thrombosis in a neonate. PMID- 7491006 TI - Human ehrlichiosis in the UK. PMID- 7491007 TI - Impact of treatment and prophylaxis on presentation and survival of patients with AIDS in Milan. PMID- 7491008 TI - HIV prevalence among pregnant women in Kigali, Rwanda. PMID- 7491009 TI - Genetic diversity and HIV detection by PCR. PMID- 7491010 TI - Laboratory-acquired Helicobacter pylori infection. PMID- 7491011 TI - Good clinical observation is essential before molecular studies. PMID- 7491012 TI - Day-time naps and melatonin in blind people. PMID- 7491014 TI - Assessing suffering. PMID- 7491013 TI - Improvement of sleep quality by melatonin. PMID- 7491015 TI - Issues for the FDA. PMID- 7491016 TI - Issues for the FDA. PMID- 7491017 TI - Midwifery in obstetrics. PMID- 7491018 TI - Midwifery in obstetrics. PMID- 7491019 TI - Midwifery in obstetrics. PMID- 7491020 TI - Low birthweight at Kasaji Hospital, Zaire. PMID- 7491021 TI - Predicted homozygous mis-sense mutation in Gilbert's syndrome. PMID- 7491022 TI - Diabetes insipidus with Behcet's disease. PMID- 7491023 TI - Behcet's disease and coeliac disease. PMID- 7491024 TI - Enzyme defect as a risk factor for spina bifida. PMID- 7491025 TI - Radiological features of fibrosing colonopathy. PMID- 7491026 TI - Is intravenous albumin a placebo? PMID- 7491027 TI - Immunosuppression induced by hepatitis C virus infection reduces acute renal transplant rejection. PMID- 7491029 TI - Heliox treatment for pneumatosis cystoides coli. PMID- 7491028 TI - Hypertension with moclobemide. PMID- 7491030 TI - Nodular gastritis associated with Helicobacter helmannii infection. PMID- 7491031 TI - Mathematical bottom-line of intraoperative haemodilution. PMID- 7491032 TI - Neural networks. PMID- 7491033 TI - Neural networks. PMID- 7491034 TI - Neural networks. PMID- 7491035 TI - Is mortality rate what it is all about in congestive heart failure? PMID- 7491036 TI - Hiking sticks in mountaineering. PMID- 7491037 TI - Tickborne encephalitis despite specific immunoglobulin prophylaxis. PMID- 7491038 TI - SIDS theory: from hype to reality. PMID- 7491039 TI - MAST-I: agreeing to disagree. Multicentre Acute Stroke Trial--Italy Group. PMID- 7491041 TI - Research in emergency care without consent: new proposed FDA rules. PMID- 7491040 TI - Thrombolytic therapy for acute ischaemic stroke: promising, perilous, or unproven? PMID- 7491042 TI - Does surgery disseminate or accelerate cancer? PMID- 7491043 TI - Preventing HIV transmission in prison: a tale of medical disobedience and Swiss pragmatism. PMID- 7491044 TI - Randomised controlled trial of streptokinase, aspirin, and combination of both in treatment of acute ischaemic stroke. Multicentre Acute Stroke Trial--Italy (MAST I) Group. AB - In ischaemic stroke, thrombolytic drugs speed the recanalisation of intracerebral arteries. The effects of aspirin are not known. A trial was conducted to determine whether, separately or together, streptokinase and aspirin have clinical benefits in acute ischaemic stroke similar to those in acute myocardial infarction. 622 patients with acute ischaemic stroke within 6 hours of symptom onset were randomised with a 2 x 2 factorial design to (i) a 1-hour intravenous infusion of 1.5 MU streptokinase, (ii) 300 mg/day buffered aspirin for 10 days, (iii) both active treatments, or (iv) neither. Early results raised a question whether the trial should be continued. Streptokinase (alone or with aspirin) was associated with an excess of 10-day case fatality (odds ratio 2.7; 95% confidence interval 1.7-4.3; 2p < 0.00001). Of the four groups randomised, only patients allocated to streptokinase plus aspirin had a significantly higher risk of early death than those given neither streptokinase nor aspirin (odds ratio 3.5; 95% CI 1.9-6.5; 2p < 0.00001). Streptokinase (alone or with aspirin) and aspirin (alone or with streptokinase) reduced, albeit not significantly, the incidence of combined six-month case fatality and severe disability: odds ratio for streptokinase 0.9 (95% CI 0.7-1.3) and odds ratio for aspirin 0.9 (95% CI 0.6 1.3). The risk of early death with thrombolytic treatments should be weighed against the potential benefit of a marginal reduction of severe disability after the first six months. PMID- 7491045 TI - Dissent: an alternative interpretation of MAST-I. Multicentre Acute Stroke Trial- Italy Group. PMID- 7491047 TI - Mortality after prostatectomy: selection and surgical approach. AB - Observational studies have found higher long-term mortality after transurethral prostatectomy (TURP) than after open prostatectomy (OP) and that this difference remained after statistical adjustment for comorbidity. This higher mortality has been attributed to the transurethral procedure itself. This association is reassessed here. Time-sequenced hospital and death records were analysed for 13,815 men undergoing prostatectomy (not for cancer) during the years 1963-85. TURP had a lower 30-day case-fatality rate than had OP, a similar 90-day rate, but TURP had a higher one-year rate. By inference, any excess mortality after TURP must begin shortly after the first postoperative month. However, plotting mortality expressed as SMRs, for the three years after operation showed no increase in long-term mortality after TURP, nor was there any concomitant increase in one-year death rates after prostatectomy as TURP replaced OP. Long term mortality after TURP was close to that expected from background population rates: after SMR for TURP, for the second and third postoperative years, was 100 (95% CI 93-107). In contrast, long-term mortality after OP was lower than expected from population rates with a corresponding SMR of 79 (95% CI 71-88). The apparent excess in long-term mortality after TURP is unlikely to be caused by the operation itself. It is more likely to reflect relatively low long-term mortality in OP patients as a consequence of OP patients having been relatively fitter than those having TURP. PMID- 7491046 TI - Toxic gas generation from plastic mattresses and sudden infant death syndrome. AB - Microbial generation of toxic gases from antimony, arsenic, or phosphorus in compounds used as fire retardants in cot mattresses has been proposed as a cause of sudden infant death. To test this hypothesis, 23 polyvinyl chloride mattress samples from cot death cases were incubated on malt agar plates until good microbial growth was obtained. Silver nitrate and mercuric chloride test papers were then inserted and the colour reactions recorded. The predominant organism, recovered from all mattresses tested, was not, as claimed in earlier work, the fungus Scopulariopsis brevicaulis, but a mix of common environmental Bacillus spp. Test paper colour changes occurred whenever bacterial growth was present, but these reactions also occurred in control tests in which no mattress material was present on the plates. Chemical and instrumental analyses of exposed test papers showed that the colour reactions were not due to deposits of antimony, arsenic, or phosphorus. Our findings do not support the hypothesis that toxic gases derived from antimony, arsenic, or phosphorus are a cause of sudden infant death. More sulphur was found in test papers exposed in plates containing bacterial growth than in those without such growth. This result suggests that the test paper reactions were due to the generation of sulphur-containing compounds during bacterial growth on the agar medium. PMID- 7491048 TI - Image analysis of beta-amyloid load in Alzheimer's disease and relation to dementia severity. AB - The protein beta-amyloid is said to be central to the disease process of Alzheimer's disease (AD). Several groups have developed transgenic models that overexpress the amyloid precursor protein or beta-amyloid and then develop AD like neuropathology. Another report suggests that beta-amyloid accumulation in old dogs correlates with cognitive impairment. However, many other researchers argue that beta-amyloid deposition in senile plaques is a secondary event because plaque numbers in man do not correlate well with cognition. We set out to analyse this conumdrum in man. We selected 16 mild to severely demented AD cases on the basis of mini-mental state exam scores (MMSE; n = 16). We also included 4 controls who represented the upper range of cognitive ability. We used a computer based image analysis of cross-sectional area of the brain occupied by beta amyloid immunopositive deposition. We used this technique in preference to conventional methods of manual plaque counts and found a strong relation between beta-amyloid load in entorhinal cortex and cognition measured on various scales (r = -0.93 versus the Blessed IMC). Our study suggests that the size of cortical area affected by beta-amyloid deposition is an important factor in the clinical manifestation of dementia, and lends support to the possibility that beta-amyloid is central to the aetiology of AD. PMID- 7491049 TI - Haematogenous dissemination of prostatic epithelial cells during radical prostatectomy. AB - Radical prostatectomy is one treatment for organ-confined prostatic adenocarcinoma. Dissemination of malignant prostatic cells after radical prostatectomy could be partly due to prostate manipulation during dissection. We confirmed by assay of prostate-specific membrane antigen by reverse-transcription nested PCR that prostate manipulation seeded prostatic epithelial cells in the general circulation in 12 of 14 consecutive patients operated on for organ confined prostate adenocarcinoma. Our results suggest that surgeons should approach radical prostatectomy with care to avoid seeding from the prostate gland. Antiandrogen therapy might reduce the haematogenous spread of prostatic cells during radical prostatectomy. PMID- 7491050 TI - Survival after early treatment for carbamyl phosphate synthetase (CPS) I deficiency associated with increase of intramitochondrial CPS I. AB - The basis for the benefit of early treatment in urea-cycle defects might be an increase in intramitochondrial mutant enzyme in hepatocytes in the postnatal period. In two siblings with carbamyl phosphate synthetase I (CPS I) deficiency, immunoreactive CPS I was greatly reduced in the liver and no residual enzyme activity was detectable. The elder child died at age 4 days, before the diagnosis of CPS I deficiency was established, but in the younger child, age 9 months, treatment was initiated on the 2nd day of life when ammonia concentration was moderately increased, and she has survived. Intramitochondrial CPS I was substantially higher in this sibling than in the elder sister. The different outcome in the younger patient was probably attributable to prompt treatment after early diagnosis. PMID- 7491051 TI - A young woman with lymphoma and endocarditis. PMID- 7491054 TI - Health bucks the squeeze on cash. PMID- 7491053 TI - Overdiagnosis of schizophrenia: role of alcohol and drug misuse. PMID- 7491055 TI - AIDS education dollars cause upset in Texas. PMID- 7491052 TI - Acute renal failure. PMID- 7491056 TI - Factors that suppress HIV uncovered. PMID- 7491057 TI - Familial herpes encephalitis. PMID- 7491058 TI - Malaria vaccine in children under 12 months of age. PMID- 7491059 TI - Malaria vaccine in children under 12 months of age. PMID- 7491060 TI - Chloroquine-resistant falciparum malaria in Mauritania. PMID- 7491061 TI - Myasthenia gravis and levonorgestrel implant. PMID- 7491062 TI - Is there a synergistic effect between previous and current vaccination against influenza? PMID- 7491063 TI - Do microbiota cause emission of stibine from cot mattresses? PMID- 7491066 TI - Epidermal mast cells in atopic dermatitis. PMID- 7491065 TI - KS-associated herpesvirus-like DNA sequences after allogeneic bone-marrow transplantation. PMID- 7491064 TI - Spurious rises of cardiac troponin T. PMID- 7491067 TI - Are assault victims prone to the surgeon's lancet (and wounds that surely heal)? PMID- 7491068 TI - Turning to primary care. PMID- 7491069 TI - Turning to primary care. PMID- 7491071 TI - Turning to primary care. PMID- 7491070 TI - Turning to primary care. PMID- 7491072 TI - National drug policies. PMID- 7491073 TI - Travellers' health and the economy of developing nations. PMID- 7491074 TI - Pure alcoholic fatty liver and progression to cirrhosis or fibrosis. PMID- 7491075 TI - Increase in thyrotoxicosis associated with iodine supplements in Zimbabwe. PMID- 7491076 TI - Effect of goitre on phospholipid fatty-acid metabolism in children. PMID- 7491077 TI - Hepatitis B immunisation in chronically transfused recipients. PMID- 7491078 TI - Hepatitis B immunisation in chronically transfused recipients. PMID- 7491079 TI - Sex chromosomal abnormalities and intracytoplasmic sperm injection. PMID- 7491080 TI - Accelerated decline in cervical cancer mortality in England and Wales. PMID- 7491081 TI - Inhaled steroids and psoriasis. PMID- 7491082 TI - Protein-losing enteropathy from cap polyposis. PMID- 7491084 TI - English as she is wrote. PMID- 7491083 TI - English as she is wrote. PMID- 7491085 TI - English as she is wrote. PMID- 7491086 TI - New imidazo[1,2-b]pyridazine ligands for peripheral-type benzodiazepine receptors on mitochondria and monocytes. AB - Several 6-chloro-2,3-disubstituted imidazo[1,2-b]pyridazines, selected from a number of synthetic imidazo[1,2-b]pyridazines which lacked significant binding activity at central benzodiazepine receptors, potently inhibit [3H]diazepam, [3H]Ro5-4864 and [3H]PK11195 binding to rat kidney mitochondrial membranes. In membrane preparations from cultures of THP-1 cells, a human monocytic leukaemia cell line, the isoquinoline carboxamide PK11195 is strongly bound but the benzodiazepine ligands, diazepam and Ro5-4864, are much more weakly bound. The imidazopyridazine compounds which bind strongly to mitochondrial benzodiazepine receptors are very potent displacers of [3H]PK11195 bound to the THP-1 membranes. It appears that the binding properties of these new imidazopyridazine ligands at 'peripheral-type' benzodiazepine receptors resemble those of the isoquinoline carboxamides more than those of the benzodiazepines. PMID- 7491087 TI - Effects of alfuzosin on urethral and blood pressures in conscious male rats. AB - This study was undertaken to determine simultaneously the effects of alfuzosin on urethral and blood pressures in the same conscious male rat. Alfuzosin (i.v., 3, 10 and 30 micrograms/kg) dose-dependently decreased urethral pressure without affecting mean arterial blood pressure. At the higher dose, blood pressure was only slightly and transiently decreased while a marked decrease (-40%) in urethral pressure was observed. Therefore, this experimental model is suitable to assess uroselectivity. PMID- 7491088 TI - Vascular alpha-1 adrenergic receptor subtypes in the regulation of arterial pressure. AB - Alpha 1 (alpha 1)-adrenoceptors can be found at numerous end organs in the autonomic nervous system, especially vascular smooth muscle. The tonic sympathetic activation of vascular alpha 1-adrenoceptors maintains vascular resistance and is vital to the regulation of arterial pressure. Recent evidence clearly demonstrates that alpha 1-adrenoceptors are a heterogenous class of receptors and that each subtype may subserve specific cardiovascular functions. Elucidation of the physiological role of each subtype in the regulation of vascular resistance and arterial pressure will enhance our understanding of the cardiovascular system and may facilitate the development of therapeutics with improved efficacy and tolerability. PMID- 7491090 TI - Adenylate cyclase activity during exocrine pancreatic proliferation in the rat. AB - Adenylate cyclase activity in pancreatic acinar cell membranes was determined in rats that had undergone a treatment with pentagastrin (250 micrograms/kg, intraperitoneal three times daily) for 1 week or that had undergone small bowel resection (90%) and were sacrified at 2 weeks, 1 month and 6 months after intervention. Both treatments are potent stimulators of pancreatic acinar cell proliferation. Adenylate cyclase activity was similar under basal conditions and after the diterpene forskolin stimulation in pancreatic acinar membranes from all groups studied. The ability of low concentrations of the stable GTP analogue, 5' guanylylimidodiphosphate (Gpp[NH]p) to inhibit forskolin-stimulated adenylate cyclase activity was decreased in pancreatic acinar membranes from enterectomized rats at 2 weeks and 1 month after the operation and returned to control values at 6 months after enterectomy. Stimulation of adenylate cyclase by high concentration of Gpp[NH]p or by secretin (10(-8) M) was higher in both pancreatic hyperplasia conditions as compared with control animals. These findings suggest that the coupling efficiency of the Gs protein to adenylate cyclase from pancreatic acinar membranes is enhanced without any alterations in the catalytic activity of the enzyme during pancreatic proliferation. In addition, it is possible that the highly regulated pancreatic acinar adenylate cyclase activity may be necessary to regulate pancreatic acinar cell proliferation. PMID- 7491089 TI - Role of nitric oxide during carrageenan-sensitized endotoxin shock in mice. AB - We have previously clarified that sensitization with a sulfated polygalactose, carrageenan (CAR), enhances endotoxin-induced tumor necrosis factor (TNF) production and lethality in mice. The present study was performed to examine the role of nitric oxide (NO) in CAR-sensitized septic mice with two different types of NO synthase (NOS) inhibitors, a non-selective inhibitor to NOS subtypes, N omega-nitro-L-arginine methyl ester (L-NAME), and a selective inhibitor to inducible NOS, aminoguanidine. Seven or eight-week-old male ddY mice were given 5 mg of CAR intraperitoneally as a primer. Then, 5 micrograms of lipopolysaccharide (LPS) was injected into the tail vein 16 hours later the pretreatment. Marked synthesis of NO was induced in CAR-sensitized mice, as indicated by the high plasma levels of the stable endproducts, NO2-/NO3- peaking at 12 hr after the LPS challenge. The peak values at 12 hr after the LPS challenge were dependent on the dose of CAR with 1 to 5 mg, although the injection with 10 mg of CAR was adversely inhibited NO production compared with 5 mg of CAR. The LPS challenge was followed by either L-NAME (0.25, 0.5 or 1 mg) or aminoguanidine (1, 2 or 4 mg) in the septic mice sensitized with 5 mg of CAR. L-NAME reduced the plasma NO2 /NO3- level in a dose-dependent fashion, although it augmented liver injury, as measured by plasma levels of ornithine carbamyltransferase (OCT) and the LPS induced lethality in a dose-dependent fashion. In contrast, aminoguanidine did not significantly deteriorate either liver injury or lethality in spite of the decrease of NO endproducts in a similar fashion to L-NAME. These findings suggest that the inhibition of constitutive NOS is detrimental and augments LPS-induced liver injury and subsequent lethality. PMID- 7491092 TI - Cerebral uptake of morphine in the pig calculated from arterio-venous plasma concentration gradients: an alternative to tissue microdialysis. AB - The aim of this study was to characterize the reversible cerebral uptake of morphine in the pig by measuring the changing arterio-venous plasma concentration gradient over the brain. Seven pigs were anaesthetized by continuous infusions of ketamine and pancuronium and ventilated with oxygen in nitrous oxide. During and after 5-min intravenous infusions of morphine hydrochloride, blood samples were drawn from a central artery and from the internal jugular vein. Concomitantly, cerebral blood flow (CBF) was repeatedly measured as clearance of 133Xe from the brain after intracarotid injection. Plasma concentrations of morphine and, in samples from two animals, morphine glucuronides were assayed by high-performance liquid chromatography. Drug flux (Jnet) from arterial blood to brain was calculated from the arterio-venous plasma concentration gradients, the blood:plasma concentration ratio and CBF. Uptake of morphine from arterial blood to brain was very rapid, with a maximal Jnet typically at 3 min after the beginning of the infusion. The initial cerebral extraction of morphine was close to 50%. When the arterial and jugular venous concentration curves crossed, 1-5 min after the end of the infusion, the initially rapid uptake of morphine changed into a slow and steady release. The cerebral extraction of morphine glucuronides was comparable to that of morphine, however, Jnet was lower due to lower plasma concentrations at time of maximal extraction. The findings demonstrate how the cerebral uptake and release of morphine and its metabolites can be studied with a method that is entirely non-invasive to the brain and permits very flexible sampling. Uptake and release of drug is observed directly and need not be inferred from cerebral concentration curves. PMID- 7491091 TI - Pancuronium masks the prejunctional muscarinic autoreceptor in guinea pig tracheal smooth muscle. AB - The effect of pancuronium pretreatment on the function of the prejunctional muscarinic receptor in guinea-pig trachea was studied by using electrical field stimulation (EFS). The effects of cumulative doses of the muscarinic M2 receptor antagonist gallamine were investigated in tracheal smooth muscle strips from guinea-pigs after addition of pancuronium in vitro and in strips from guinea-pigs which had been pretreated with doses of pancuronium that caused 100% neuromuscular blockade. The results of both types of experiments were compared to those of control groups of the same size. In all strips a dose response curve with cumulative doses of methacholine was made before EFS was switched on. No differences were found between the mean pD2 value and slope of the concentration response curves of untreated guinea-pigs and animals treated with anaesthetics and pancuronium. The animals showed variable responses to pancuronium. The bath concentration of pancuronium which decreased the EFS-induced contraction to half the original value varied between 14-61 microM. The intravenous dose necessary to paralyze the muscles, varied among the different guinea-pigs from 0.017-0.085 mg.kg-1. The EFS-induced contraction for the concentration range of gallamine 0.32 microM-0.32 mM was found to differ significantly between the strips treated with pancuronium in the organ bath and their control group. For the guinea-pigs anaesthetized and pretreated with pancuronium a significant difference with control was observed at gallamine concentrations ranging from 0.032-0.32 mM. These results show that pancuronium, added to the organ bath as well as administered intravenously to the guinea-pig, masked the inhibitory muscarinic receptor. PMID- 7491093 TI - Antagonist of N-methyl-D-aspartate receptors partially prevent the development of cocaine sensitization. AB - Behavioral sensitization to cocaine was tested for in rats pretreated with MK 801, a noncompetitive N-methyl-D-aspartate (NMDA) receptor antagonist, or D-3-(2 carboxypiperazin-4-yl)-1-propenyl-1-phosphonic acid (D-CPPene), a competitive NMDA antagonist. A 5-day regimen of once-daily cocaine (15 mg/kg) injections yielded sensitization to cocaine (15 mg/kg)-induced behavioral activation. Cocaine sensitization was partially prevented by MK-801 (0.25 mg/kg) or D-CPPene (20 mg/kg) pretreatment. These results differ from previous reports that NMDA receptor antagonists completely prevented the development of stimulant sensitization. While raising questions about methodological differences among laboratories studying this issue, our findings suggest that sensitization may involve mechanisms dependent on NMDA-receptor function as well as NMDA receptor independent mechanisms. PMID- 7491094 TI - PK 11195 reduces the brain availability of lindane in rats and the convulsions induced by this neurotoxic agent. AB - The effect of pretreatment with PK 11195, a ligand of the 'peripheral-type' benzodiazepine receptor (PBR), on convulsions induced by lindane (gamma hexachlorocyclohexane, gamma-HCH) in rats was examined, to determine whether the mechanism of this convulsant activity may be mediated through the PBR. PK 11195 elicited a protective effect against the convulsant activity of orally administered lindane. It reduced the frequency of animals exhibiting convulsions and delayed the time to onset of these seizures. The concentration of lindane in the brain was found to be significantly lower in PK 11195 pretreated rats and a high correlation between blood and brain lindane concentrations was obtained. When similar experiments were repeated with alpha-HCH, a non-convulsant isomer of HCH, brain and blood concentrations were again found to be significantly reduced in PK 11195 pretreated animals. We conclude that the 'anticonvulsant' action of PK 11195 was not due to an interaction of PK 11195 and lindane on common CNS target sites, but by an action of PK 11195 on the gastrointestinal tract of the animal, delaying the absorption of lindane into the bloodstream. PMID- 7491095 TI - Muscarinic agonist-induced positive inotropic response in chick atria. AB - Carbachol (10(-6)-3X10(-4)M) induces a positive inotropic response in paced, pertussis toxin-treated fibers which is atropine-sensitive and independent of endogenous catecholamines. At the same concentrations in atria from saline treated chicks, carbachol's negative inotropic effect on the steady state contractions (SSC) is attenuated and the rested state contraction (RSC) is increased. The RSC and SSC in pertussis toxin-treated fibers are increased by carbachol (EC50 = 30 microM) indicating that repetitive electrical depolarization is not essential for the inotropic response. The inotropic response of the SSC is frequency-independent from 0.10-1.0 Hz; however it is decreased (approximately 50%) at a high frequency (3.0 Hz). In control untreated atrial muscle, carbachol (10(-4)M) selectively increases the early component of the RSC. The late component of the RSC, representing activation of transmembrane Ca2+ inward current, is not changed. Carbachol's positive inotropic effect is perhaps exerted by enhancing Ca2+ release and/or Ca2+ content of the sarcoplasmic reticulum. The ability of various muscarinic agonists to induce a positive inotropic response was: carbachol > acetylcholine > oxotremorine. This order correlates with the ability of these agents to induce a tetrodotoxin-resistant Na+ inward current that increases intracellular Na+ and to promote phosphatidylinositol hydrolysis. These data are consistent with the hypothesis that the carbachol-induced positive inotropic response may result from greater intracellular Ca2+ availability secondary to enhanced Na-Ca exchange. The greater Ca2+ availability, together with increased production of the Ca-mobilizing messenger, inositol 1,4,5 trisphosphate (InsP3), can exert a synergistic effect to regulate force generation. PMID- 7491096 TI - Selective potentiation of IGE-dependent histamine release from rat peritoneal mast cells by stem cell factor. AB - Effect of stem cell factor on histamine release from rat peritoneal mast cells was studied. Although stem cell factor did not evoke histamine release by itself, it clearly potentiated histamine release from sensitized mast cells caused by antigen, anti-IgE and concanavalin A. However, stem cell factor did not affect histamine release caused by compound 48/80, calcium ionophore A23187 and substance P. Although maximum potentiation of antigen-induced histamine release by stem cell factor was accomplished after 1-10 minute-preincubation, potentiation was decline after a longer incubation period. Potentiation of histamine release by phosphatidylserine and non-mast cells in the rat peritoneal cavity was incubation time-dependent. Potentiation by stem cell factor was additive to that by phosphatidylserine or non-mast cells. These results indicate that stem cell factor selectively potentiates IgE-dependent histamine release from rat peritoneal mast cells, and suggest that the mechanism involved is distinct from that of phosphatidylserine or non-mast cells in the rat peritoneal cavity. PMID- 7491098 TI - A strategic framework for infant mortality reduction: implications for "Healthy Start". AB - The high infant mortality rate in the United States, especially in urban areas, remains a major federal concern. Four strategies for reducing infant mortality in cities participating in the federal ?Healthy Start? are reducing high-risk pregnancies; reducing the incidence of low birthweight and preterm births; improving birthweight-specific survival; and reducing specific causes of post neonatal mortality. Estimates of the impact of known interventions indicate that the reduction in infant mortality would be large for only one strategy: improving birthweight-specific survival. Most interventions yield a 2 percent reduction, or less, in mortality and when combined, would amount to about 30 percent. This strategic model provides a realistic framework to assess the impact of the Healthy Start demonstration and is useful in highlighting the interventions most likely to reduce infant mortality in a population. PMID- 7491097 TI - The institutional review board and beyond: future challenges to the ethics of human experimentation. AB - Over the past two decades, institutional review boards (IRBs) have transformed the conduct of clinical research, in the process protecting human subjects and setting an admirable standard for monitoring the ethics of science. Nevertheless, the very proliferation of these committees, in addition to changing the character and sponsorship of new research, suggests that a ?one size fits all? approach to the governance of human experimentation may have outlived its usefulness. It may be time to remove the ?I? from the IRB and create a system with greater national oversight. Whether such a change can be accomplished within the current political climate is debatable. But the need for such a shift is becoming increasingly apparent. PMID- 7491099 TI - Local responses to expanded Medicaid coverage for pregnant women. AB - Concern about high infant mortality and morbidity in the United States, combined with the erosion of private insurance coverage, sparked major expansions in the Medicaid program in the 1980s. This study examines how the Medicaid expansions for pregnant women affected access to prenatal care for low-income women through case studies conducted in four states early in 1991. Despite the significantly greater share of births covered by Medicaid in the period 1986 to 1991, the timely initiation of prenatal care improved in only one state. Although prenatal services increased in some areas, significant problems persisted in others. The growth in capacity of the prenatal care system was greatest when state and local policies designed to increase supply were also instituted. While the Medicaid expansions eliminated significant barriers to prenatal care for low-income women, other policies that have been designed to reduce the remaining barriers may be necessary in order significantly to expand access to prenatal care and to improve birth outcomes. PMID- 7491100 TI - Rethinking the organization of children's programs: lessons from the elderly. AB - The U.S. system of care for children is a collection of activities and funding mechanisms that create a complex, fragmented patchwork of services and programs. In contrast, the elderly enjoy universal entitlement to national health insurance through Medicare, a uniform level of income security, and an organized system of community-based services. The provisions of the Older Americans Act (OAA) are compared with the current program of maternal and child health services. Basic features of the national policy--and the core program and administrative infrastructure of the OAA--may serve as a model to meet the special challenges of child and family services. The tools for building the coordinated, comprehensive service system that was devised for the elderly are also available to improve the lives of children. Federal legislation is needed to create the special protections that are basic to the organization and delivery of services and to the assurance of a permanent national policy focus for children and families. PMID- 7491101 TI - The cancer burden of southern-born African Americans: analysis of social geographic legacy. AB - Southern-born black Americans, especially those who migrated to the Northeast and Midwest, had much higher cancer mortality rates during the period 1979 to 1991 than their counterparts who were born and died outside the South. Elevated rates were apparent for the 35- to 44-year-old age group, and were highest among the elderly. The largest and most consistent differences between Southern-born and Northeastern, Midwestern, and Western-born African Americans were for cancer of the breast (female), esophagus, larynx, and lung (male), pancreas, prostate, and stomach. The combined effects of nutritional imbalances, cigarette smoking, high risk jobs, limited access to medical screening and care, and other factors associated with poverty are suggested as etiologic factors common to the high risk, Southern-born black population. It is also possible that the Southern-born excess of cancer deaths is at least partly an artifact of the data and the ecological level of the analysis. PMID- 7491102 TI - Cancer prevention strategies for the future: risk identification and preventive intervention. AB - The knowledge base may be advanced by concentrating on primary prevention of cancer, which in turn will inform the development of effective and efficient intervention strategies for prevention that may substantially reduce cancer incidence. Several priority areas have been identified, and with new multidisciplinary approaches spanning the biological, epidemiological, social, behavioral, and economic sciences, new initiatives in primary cancer prevention may lead the way toward reducing cancer mortality through primary prevention. PMID- 7491104 TI - Repression of the ovalbumin gene involves multiple negative elements including a ubiquitous transcriptional silencer. AB - Most eukaryotic genes are controlled by a complex array of cis-acting regulatory elements that modulate transcriptional activity. Two major regulatory elements reside in the chicken ovalbumin gene, a steroid-dependent regulatory element (SDRE, -892 to -780) and a negative regulatory element (NRE, -308 to -88). The SDRE is required for responsiveness to estrogen and glucocorticoid. The NRE appears to have the dual role of repressing transcription in the absence of steroids and of cooperating with the SDRE to activate transcription in the presence of steroids. The experiments described herein were designed to investigate the role of the NRE in repressing gene expression. Transfection of OvCAT fusion genes containing deletions in the NRE into primary oviduct cell cultures identified three elements (-308 to -256, -239 to -220, and -174 to -88) that repress transcription. Oligomers corresponding to portions of these elements also independently repress the viral thymidine kinase promoter. Interestingly, the element from -239 to -220 functions mechanistically as a silencer and shares sequence identity with silencers in other genes (TCTCTCCNA). Mobility shift studies indicated that all of the negative elements bind specific protein complexes from oviduct, none of which is appreciably affected by treatment with steroid hormones. However, oviduct-specific proteins bind to the regions from 280 to -252 and from -134 to -88, providing the first identification of potential tissue-specific elements in the ovalbumin gene. These results demonstrate that the region of DNA originally called the NRE is a multifunctional regulatory element that may be involved in several diverse regulatory activities. PMID- 7491103 TI - The general hospital and mental health care: a British perspective. AB - For over a century, the specialist mental hospital was the basis of psychiatric care in all industrialized countries. Britain was among the first to establish national requirements for asylum care, although these institutions were financed and managed locally. Psychiatric treatment in general hospitals began in the later eighteenth century, but, except for some outpatient clinics, this practice ended prematurely in the mid-nineteenth century, not to resume until after World War II. The National Health Service (NHS) provided the framework within which general hospital psychiatry was re-created, forming the nucleus for district services. Although the intent of NHS reform in the 1980s was to establish a reduced market hospital system that would continue to offer long-term care, this service seems likely to continue for only a few more years. ?Reforms? in the NHS and social services have placed the whole of British mental health care in a situation of uncertainty. PMID- 7491106 TI - Negative regulation of chicken GATA-1 promoter activity mediated by a hormone response element. AB - GATA-1 is a DNA-binding protein that regulates transcription of erythroid specific genes and is required for the formation of mature erythroid cells. We show here that the GATA-1 hormone response-like element (GHRE) within the first intron of the gene functions as an inhibitory element in chicken erythroid precursor cells, as revealed by expression studies with mutants of the minimal GATA-1 promoter. We identify in these precursor cells the relevant proteins that interact with GHRE as a heterodimer of the thyroid hormone receptor alpha and the chicken ovalbumin upstream promoter transcription factor. Our results indicate that this novel complex can negatively regulate the GATA-1 promoter and suggest that GATA-1 can overcome this inhibitory action. We provide evidence that the viral gene product, v-erb A, can also reduce GATA-1 promoter activity through the GHRE site. PMID- 7491105 TI - The reduction of insulin gene transcription in HIT-T15 beta cells chronically exposed to high glucose concentration is associated with the loss of RIPE3b1 and STF-1 transcription factor expression. AB - The most important regulator of insulin gene expression in pancreatic beta- cells is glucose, which affects gene transcription, mRNA translation, and secretion. Insulin gene transcription is both positively and negatively regulated by glucose. Recently, we have shown that the inhibition of insulin gene transcription caused by passaging HIT T-15 beta-cells, in the presence of high glucose, was due, in part, to reduced expression of a key regulator of insulin enhancer-mediated expression, somatostatin transcription factor-1 (STF-1). In this study, we have examined whether the activity of the other essential transcription regulators of insulin gene expression, the RIPE3b1 and insulin control element (ICE) activators, were also influenced in these HIT T-15 cells. The results show that the binding and trans-activation functions of the RIPE3b1 activator are reduced in parallel with the loss in STF-1 and insulin gene expression. In contrast, the regulatory properties of the ICE activator are unaffected. Our studies indicate that insulin gene transcription is inhibited by glucose through a mechanism involving reduced expression of both the RIPE3b1 and STF-1 activators in HIT T-15 cells but is independent of the ICE activator. PMID- 7491107 TI - Regulation of insulin-like growth factor I receptor gene expression by Sp1: physical and functional interactions of Sp1 at GC boxes and at a CT element. AB - The insulin-like growth factor I (IGF-I) receptor mediates signal transduction by the IGFs and plays a critical role in growth and development. The proximal promoter region of the rat IGF-I receptor gene contains multiple Sp1 consensus binding sites (GC boxes). Various promoter fragments fused to a luciferase reporter gene were transiently cotransfected together with an Sp1 expression vector into Drosophila Schneider cells, which lack endogenous Sp1. A proximal promoter fragment containing 476 nucleotides of 5'-flanking region and 640 nucleotides of 5'-untranslated region was strongly activated by Sp1 (an average of 116-fold), and progressive 5'-deletions of the promoter that sequentially removed GC boxes reduced Sp1 activation to 15-fold over basal promoter activity. DNase I footprinting studies with purified Sp1 protein revealed four GC boxes in the 5'-flanking region of the promoter and one homopurine/homopyrimidine motif (CT element) in the 5'-untranslated region that bound Sp1. Mutation of the CT element reduced Sp1 activation by 70%. Taken together, these results demonstrate that Sp1 can regulate expression of the IGF-I receptor promoter by acting both on GC boxes in the 5'-flanking region of the promoter and on a CT element in the 5' untranslated region. PMID- 7491108 TI - Insulin-like growth factor-I and epidermal growth factor interact to regulate growth and gene expression in IEC-6 intestinal epithelial cells. AB - Epidermal growth factor (EGF) and insulin-like growth factor-I (IGF-I) exert trophic effects on bowel mucosa. Each growth factor uses a distinct tyrosine kinase receptor but the receptors share some common signal transduction pathways. In other systems, regulation of cell growth involves interactions among multiple growth factors. We used IEC-6 cells, an epithelial cell line established from rat small intestine, to test whether EGF and IGF-I interact to regulate intestinal epithelial cell growth. EGF and IGF-I alone each stimulated DNA synthesis in IEC 6 cells. EGF was more potent than IGF-I, and effects of the two growth factors in combination were synergistic. Characterization of the IGF system [IGF-I, IGF-II, type 1 IGF receptor, and six IGF binding proteins (IGFBPs) 1-6] revealed that IEC 6 cells express high levels of type 1 IGF receptor mRNA, low or undetectable levels of IGF-I and IGF-II mRNAs, and mRNA for only one of the six IGFBPs, IGFBP2. IGF-I decreases expression of type 1 IGF receptor mRNA in IEC-6 cells and EGF attenuates this effect. EGF and IGF-I both reduce IGFBP2 mRNA expression, and inhibitory effects of EGF and IGF-I in combination are additive. EGF reduces IGFBP2 accumulated in conditioned medium relative to levels observed with IGF-I alone. These effects of EGF on type 1 IGF receptor expression and on levels of IGFBP2 mRNA and IGFBP2 in medium may contribute to synergistic mitogenic effects with IGF-I by promoting IGF-I responsiveness.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7491109 TI - A highly conserved region in the hormone-binding domain of the human vitamin D receptor contains residues vital for heterodimerization with retinoid X receptor and for transcriptional activation. AB - Residues located between amino acids 244 and 263 in the human vitamin D receptor (hVDR) show extensive homology with other members of the steroid/thyroid/retinoid hormone receptor superfamily. The corresponding region of the glucocorticoid receptor has been shown to interact with the 90-kilodalton heat shock protein (hsp90), yet hVDR does not appear to bind to hsp90. Herein we report a study of hVDR in which the functional role of five conserved residues was tested by replacing Phe-244, Lys-246, Leu-254, Gln-259, and Leu-262 with glycines by site directed mutagenesis. Initial screening of these mutants indicated that all were significantly impaired in their ability to activate transcription from a vitamin D-responsive reporter construct when expressed in transfected VDR-deficient COS-7 cells. Further characterization revealed two classes of mutants: the predominant class binds the 1,25-dihydroxyvitamin D3 ligand normally but is defective in its ability to form a heterodimeric complex with the retinoid X receptor (RXR) on a vitamin D responsive element (VDRE). A second unique class, represented by a single mutant at Lys-246, is normal both with respect to ligand binding and complex formation but still very impaired in transactivation ability. The distinction between these two classes was confirmed by the demonstration that a member of the first class, with a mutation at Gln-259, could be restored to near wild type transactivation ability by supplying excess RXR, while the Lys-246 mutant could not be so rescued. We therefore conclude that the primary function of this conserved domain in hVDR is the mediation of heterodimerization with RXR, leading to VDRE binding and transactivation. The possibility also exists that the Lys-246 mutant may be impaired in a step of transactivation that is distal to complex formation with RXR on the VDRE, perhaps in interactions with the transcriptional machinery itself. PMID- 7491110 TI - Role of the TATA box in transcription of the mouse mammary tumor virus long terminal repeat. AB - An in vitro transcription system from mammary cells was established to study transcription of the long terminal repeat (LTR) of the mouse mammary tumor virus (MMTV). Experiments with progressive 5'-deletion constructs of the MMTV LTR revealed that a 19-base pair (bp) region from -41 to -23 bp, encompassing the TATA box and flanking DNA sequence, was as transcriptionally active as larger promoter constructs, both in nuclear extracts from human mammary cell lines (T47D and MCF7) and a nonmammary cell line (HeLa). The cell-free system was capable of supporting transcriptional induction by factors binding upstream of the TATA box, however, since purified glucocorticoid receptor-induced transcription in larger promoter constructs encompassing the MMTV hormone-responsive elements. Transcription from two other promoters, the adenovirus major late promoter and the human immunodeficiency virus LTR, also revealed a significant transcriptional contribution of upstream elements. The 19-bp TATA region from the MMTV LTR was shown to have considerably more activity in this transcription system than comparable TATA regions from other promoters. Sequences critical to the MMTV TATA region were evaluated by single base pair mutagenesis and found to comprise a consensus TATA box sequence, TATAAAA, as well as a single A just upstream of the TATAAAA sequence. Thus, the high level of basal transcription observed with the TATA region from MMTV is due to a perfect consensus TATA box sequence and a single base immediately 5' adjacent. It is likely that the high basal rate of transcription observed with this TATA box region on histone-free templates represents an inappropriate level of basal expression and that a complete evaluation of transactivation mechanisms in this system will require the recapitulation in vitro of the chromatin-mediated repressive state that exists in vivo. PMID- 7491111 TI - Glucocorticoid receptor structure and function in an adrenocorticotropin secreting small cell lung cancer. AB - ACTH secretion by tumors of nonpituitary origin is characteristically resistant to negative feedback regulation by glucocorticoids. One possible mechanism for the phenomenon could be a structural defect in the intracellular glucocorticoid receptor (GR). We studied the GR in DMS-79 cells derived from a human ACTH secreting small cell lung cancer. Compared with control cells, DMS-79 cells were found to have greatly diminished GR ligand-binding activity and immunoreactive 94 kilodalton (kDa) GR content. Northern blot analysis revealed expression of GR transcripts that appeared to be slightly larger than those in control cells. A DMS-79 cell GR cDNA was cloned by reverse transcription/polymerase chain reaction amplification of mRNA using primers specific for full-length normal GR. The derived sequence of this full-length GR differed from the reported sequence by a single altered codon (G to A; Asn to Ser at codon 363) outside the steroid binding domain. This N363S DMS-79 GR functioned normally to activate a target gene [mouse mammary tumor virus-chloramphenicol acetyl transferase (MMTV-CAT)] in transient transfection experiments in COS cells. Evidence for expression of a second type of GR mRNA was obtained by screening a DMS-79 cell cDNA library. This GR cDNA contained normal GR sequence up to nucleotide 2155, corresponding exactly to the end of exon 7 in the normal GR gene. The sequence appended to the GR sequences was not matched by any known sequence in DNA databases and included an in-frame termination codon after only 6 bases. The predicted truncated GR protein product (GR delta) has a mol wt of 73,740 and lacks most of the ligand-binding domain. Transient transfection of the GR delta form into COS cells did not reveal any dominant negative effect on the function of a cotransfected normal GR.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7491113 TI - The mouse 3 beta-hydroxysteroid dehydrogenase multigene family includes two functionally distinct groups of proteins. AB - The enzyme 3 beta-hydroxysteroid dehydrogenase (3 beta HSD) plays an essential role in the biosynthesis of all steroid hormones. We previously reported the isolation, characterization, and tissue-specific expression of four distinct but highly homologous 3 beta HSD cDNAs (forms I, II, III, and IV). Enzymatic characterization of three of these isoforms demonstrated that mouse 3 beta HSD I and III function as dehydrogenase/isomerases, but 3 beta HSD IV functions exclusively as a 3-ketosteroid reductase. We now report the isolation and characterization of an additional distinct mouse 3 beta HSD cDNA, 3 beta HSD V, which is expressed in the liver of male mice beginning in late puberty. Similar to 3 beta HSD IV, 3 beta HSD V functions exclusively as a 3-ketosteroid reductase converting an active androgen, dihydrotestosterone (DHT), into an inactive androgen, 5 alpha-androstane-3 beta,17 beta-diol. Expressed 3 beta HSD V, however, exhibits a considerably lower apparent Michaelis-Menten constant (Km) value for DHT than 3 beta HSD IV (0.47 microM vs. 2.2 microM, respectively). The complete predicted amino acid sequence of 3 beta HSD II is also reported. The predicted amino acid sequence of mouse 3 beta HSD V reveals that this new form is more closely related to the 3-ketosteroid reductases, mouse 3 beta HSD IV and rat III (93 and 84% identity, respectively), than to the other rodent isoforms that share less than 75% identity.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7491112 TI - N-terminal variants of thyroid hormone receptor beta: differential function and potential contribution to syndrome of resistance to thyroid hormone. AB - The human syndrome of resistance to thyroid hormone (RTH) is associated with dominant mutations in the thyroid hormone receptor beta (TR beta) gene that generate mutant receptors with impaired binding for T3. Although the TR beta gene differentially expresses two N-terminal variant receptors, TR beta 1 and TR beta 2, functional analyses of RTH mutants have focused exclusively on TR beta 1. Since TR beta 2 is expressed in tissues that are malfunctional in RTH, the role of mutations in the context of TR beta 2 was examined. We compared the functional properties of corresponding RTH mutations in the common C-terminal domain of both TR beta 1 and TR beta 2. Wild type TR beta 1 and TR beta 2 bound similarly as homodimers and as heterodimers with retinoid X receptors to T3-responsive elements consisting of a direct repeat with 4-base pair spacing or an everted repeat. Homodimers, but not monomers or heterodimers, of both receptor subtypes were dissociated by the addition of T3. However, TR beta 2 formed at least 10 fold more stable homodimers than TR beta 1 on a palindromic repeat element, indicating that the N termini of TR beta 1 and TR beta 2 differentially influence dimerization on DNA. The RTH-like mutants of both TR beta 1 and TR beta 2 were equally insensitive to T3. They were defective in T3 binding but still bound DNA like their wild type counterparts except that the T3-dependent dissociation of homodimers from DNA was severely reduced. Wild type TR beta 1 and TR beta 2 mediated T3-inducible transactivation in cotransfection assays; this, however, was abolished in both mutants. TR beta 1 mediated more sensitive T3-dependent transcriptional suppression than TR beta 2 through the negative T3 response region of the TSH beta gene. Again, the mutation abolished T3-dependent suppression by both mutants. Furthermore, both mutants inhibited T3-inducible transcriptional activation by different wild type TR alpha and beta variants. These results indicate that both mutants have the potential to contribute to the pathogenesis of RTH and suggest that a reassessment of previous models of RTH is required to take into account the inhibitory activity of both TR beta 2 and TR beta 1 mutants. PMID- 7491114 TI - Developmentally and hormonally regulated CCAAT/enhancer-binding protein isoforms influence beta-casein gene expression. AB - A highly conserved CCAAT/enhancer-binding protein (C/EBP)-binding site centered around -134 relative to the transcription start site in the rat beta-casein gene promoter is capable of interacting specifically with recombinant and mammary gland C/EBP proteins. Western blot analysis indicates that C/EBP levels change dramatically throughout mammary gland development. C/EBP alpha expression is barely detectable in mammary glands from virgin and pregnant animals but is expressed at high levels during lactation and at lower levels during involution. The expression of three C/EBP beta isoforms [the liver-enriched activating proteins (LAPs); and the liver-enriched inhibiting protein (LIP)] is elevated throughout pregnancy, with LIP expression increasing more than 100-fold. Thus, during pregnancy, a low LAP/LIP ratio (< 5) is maintained. C/EBP beta expression decreases at parturition, with LIP diminishing to levels observed in the virgin gland. Therefore, during lactation a more than 100-fold increase in the LAP/LIP ratio is observed. Treatment of the HC11 mammary epithelial cell line with hydrocortisone results in a 10- to 20-fold inhibition of LIP expression, with only minor changes in LAP levels. Therefore, glucocorticoids may impinge upon beta-casein gene expression by altering the ratio of the inhibitory to the activating isoforms of C/EBP beta. Several previously defined casein gene promoter regions capable of conferring hormone and extracellular matrix inducibility to reporter genes in mammary cells are suggested to be composite response elements, containing putative binding sites for the same set of hormonally and developmentally regulated factors: C/EBP, MGF/Stat5, and the glucocorticoid receptor. PMID- 7491115 TI - Steroidogenic factor 1 is the essential transcript of the mouse Ftz-F1 gene. AB - Targeted disruption of the mouse Ftz-F1 gene, which encodes the orphan nuclear receptors steroidogenic factor 1 (SF-1) and embryonal long terminal repeat binding protein (ELP), established that this gene is essential for development of the primary steroidogenic tissues and for male sexual differentiation. Associated with these dramatic developmental abnormalities, all Ftz-F1-disrupted mice died in the immediate postnatal period and had very low glucocorticoid levels. In this report, we show that treatment with corticosteroids markedly prolonged survival of the Ftz-F1-disrupted mice, proving that steroid hormone deficiency causes their death. We also generated SF-1-specific knockout mice with a targeting construct that specifically disrupted the SF-1 coding sequence without impairing the ELP protein. The phenotype of the SF-1-specific knockout mice was indistinguishable from that observed in Ftz-F1-disrupted mice that lack both SF-1 and ELP. Taken together, these results indicate that SF-1 is the Ftz-F1-encoded protein that is required for multiple aspects of endocrine development and for postnatal survival. PMID- 7491116 TI - Mutational analysis of the cytoplasmic tail of the G protein-coupled receptor for parathyroid hormone (PTH) and PTH-related protein: effects on receptor expression and signaling. AB - The present studies were undertaken to examine the role of the cytoplasmic tail of the G protein-coupled receptor for PTH and PTH-related protein (PTHrP) on receptor signaling and expression. The wild type (WT) receptor (585 amino acids) and five truncated receptors whose cytoplasmic tails terminated at residues 507, 494, 474, 466, and 458 were expressed in COS-7 cells. Based on [125I]PTHrP binding, mutants T507, T494, and T466 displayed progressively decreased levels of expression, compared with WT. The tailless mutant T458 was not expressed in a functional form, whereas T474 was expressed at a level similar to WT. Comparable results were obtained when expression levels of WT and mutated PTH/PTHrP receptors were evaluated by Western blotting. Binding affinities were similar for all mutated receptors (IC50 = 1-2 nM). Immunocytochemistry showed that WT and mutated receptors were diffusely distributed, presumably at the cell surface, except for the tailless mutant T458, which displayed striking perinuclear localization. T458 did not display an adenylyl cyclase response to PTH, while the other mutants were similar to WT both with respect to their maximal adenylyl cyclase responses to PTH and to their EC50 values. Cai2+ signaling properties of these mutants were assessed as PTH-stimulated 45Ca efflux from Xenopus oocytes that had been injected with in vitro transcribed PTH/PTHrP receptor cRNAs. The WT and mutated receptors (except for T458) responded to PTH with significant (6- to 27-fold) increases in 45Ca efflux.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7491118 TI - Relaxing and unwinding on Holliday: DNA helicase-mediated branch migration. PMID- 7491119 TI - Induction and repair of (6-4) photoproducts in normal human and xeroderma pigmentosum variant cells during the cell cycle. AB - The reduced rate of (6-4) photoproduct repair observed in some cell lines may represent a more severe repair deficiency in some cohort of the cell cycle, such as S-phase. Radioimmunoassay was used to determine the kinetics of (6-4) photoproduct repair in normal human fibroblasts and xeroderma pigmentosum variant cells fractionated into different phases of the cell cycle by counterflow centrifugal elutriation. Ultraviolet fluence response curves indicated that the same amount of (6-4) photoproduct damage was induced at all phases of the cell cycle. The extent of (6-4) photoproduct repair in asynchronous XP variant cells was significantly reduced compared to normal human cells. However, the rate and extent of (6-4) photoproduct repair was constant throughout the cell cycle in both normal and XP variant cells. Hence, the UV hypersensitive and hypermutable phenotypes observed in XP variant cells are not attributable to cell cycle dependent deficiencies in excision repair nor the yield of photodamage through the cell cycle. PMID- 7491117 TI - Alternatively spliced human type 1 angiotensin II receptor mRNAs are translated at different efficiencies and encode two receptor isoforms. AB - The peptide hormone angiotensin II (AngII) plays a principal role in regulating blood pressure and fluid homeostasis. Most of its known effects are mediated by a guanine nucleotide-regulatory protein (G protein)-coupled receptor pharmacologically defined as the type-1 AngII receptor or AT1. Characterization of cDNA and genomic clones shows that the human AT1 gene contains five exons and encodes two receptor isoforms as a result of alternative splicing. Exon 5 contains the previously characterized open reading frame for AT1, and exons 1 to 3 are alternatively spliced upstream of it to generate several mRNA species, while transcripts containing exon 4 are of minor abundance. In an in vitro translation system, the presence of exon 1 was found to be extremely inhibitory to translation, probably because it can form a stable secondary structure at the RNA level. The alternatively spliced second exon also had a strong inhibitory effect on translation, presumably because it contains a minicistron commencing with an ATG in an optimal context for translation initiation. Exon 2 was similarly inhibitory to protein production in transfected cells, but exon 1 was found to enhance protein synthesis in this system. Transcripts containing exon 3 and 5, which comprise up to one-third of AT1 mRNAs in all tissues examined, encode a receptor with an amino-terminal extension of 32-35 amino acids. These transcripts were translated into a larger receptor isoform in vitro and produced a functional receptor with normal ligand binding and signaling properties in transfected cells. PMID- 7491120 TI - The yeast RAD2, but not RAD1, gene is involved in the transcription-coupled repair of thymine glycols. AB - Transcription-coupled repair of H2O2-induced thymine glycols and UV-induced pyrimidine dimers has been shown to occur in the yeast Saccharomyces cerevisiae. In order to determine whether the preferential repair of thymine glycols is carried out by the same nucleotide excision repair complex that removes pyrimidine dimers, we examined the repair of thymine glycols in two yeast mutants, rad1 delta and rad2 delta, in which the nucleotide excision repair pathway was disrupted. We find that in both wild-type and a rad1 delta mutant, repair of thymine glycols occurs faster on the transcribed strand of the GAL7 gene than on the nontranscribed strand. This indicates that transcription-coupled repair can occur in the absence of nucleotide excision repair and that repair of oxidative DNA damage initiated by an N-glycosylase can be coupled to transcription. In contrast, the initial rate of repair of thymine glycols on the transcribed strand of the GAL7 gene in a rad2 delta mutant is significantly slower than that for the wild-type cells, with kinetics similar to that of the nontranscribed strand. However, by 60 min post-treatment, the amount of repair on the transcribed strand in the rad2 delta eventually reaches that of the wild-type cells. We conclude that repair of oxidative DNA damage, such as thymine glycols, can be coupled to transcription and that RAD2 facilitates transcription-coupled repair of oxidative DNA damage in yeast. PMID- 7491121 TI - Enhanced host cell reactivation capacity and expression of DNA repair genes in human breast cancer cells resistant to bi-functional alkylating agents. AB - Human breast carcinoma (MCF7-MLNr) cells resistant to the bifunctional drugs L phenylalanine mustard (L-PAM, 5-fold resistance), mechlorethamine (9-fold), cisplatin (3-fold), and BCNU (3-fold) were used to investigate the role of DNA repair in the development of resistance to alkylating agents. We have previously shown that neither L-PAM transport and metabolism nor glutathione-associated enzymes were altered in MCF7-MLNr cells, compared to the sensitive cells MCF7-WT. This study shows that treatment of pRSV-CAT plasmid with L-PAM at concentrations up to 1 microM proportionally inhibit the expression of chloramphenicol acetyl transferase (CAT) activity, while higher concentrations abolished CAT activity. pRSV-CAT reactivation was significantly increased when plasmid was transfected into MCF7-MLNr cells, compared to MCF7-WT cells. This indicates that resistant cells have more efficient capacity to recognize and repair L-PAM induced DNA damage. The mRNA expression of DNA nucleotide excision repair genes ERCC1, XPD (ERCC2), XPB (ERCC3), and polymerase beta was found to be similar in both the MCF7-WT and MCF7-MLNr cells. Western blot analysis also reveals no difference in the expression of ERCC1, AP endonuclease, poly (ADP-ribose) polymerase, and alkyl N-purine-DNA glycosylase proteins. The lack of correlation between enhanced host cell reactivation capacity in resistant cells, and the expression of these specific DNA repair genes suggests that proteins encoded by these genes are not rate limiting steps for resistance to bi-functional alkylating drugs in human breast cancer cells. PMID- 7491122 TI - Expression in Escherichia coli of a rat cDNA encoding an apurinic/apyrimidinic endonuclease. AB - A rat cDNA (rAPEN) with 85% DNA identity to the major human apurinic/apyrimidinic (AP) endonuclease gene was used to construct a fusion between it and glutathione S-transferase (GST). The GST-rAPEN fusion was subsequently overexpressed in Escherichia coli, purified on glutathione-agarose affinity columns, and the purified protein tested for AP endonuclease activity. DNA nicks were found to be specifically introduced into AP DNA in a reaction that was dependent upon the time of incubation and the amount of GST-rAPEN added. The DNA scissions produced by GST-rAPEN were determined to be adjacent and 5' to an AP site. The purified fusion protein was also able to efficiently remove 3'-(4 hydroxy-5-phospho-2 pentenal) residues, and to a lesser extent 3'-phosphoglycolate residues. The GST rAPEN activity failed to exhibit any 3'-5' exonuclease activity, a characteristic shared by the major AP endonuclease in bovine and human. PMID- 7491123 TI - Mutagenicity, carcinogenicity and teratogenicity of lithium compounds. AB - This paper reviews the information available concerning the mutagenic, teratogenic and carcinogenic effects of lithium. Such effects would be highly unlikely in an occupational setting but might be a risk to the considerable percentage of the population treated for manic-depressive disorders. It is concluded that lithium compounds have no significant clastogenic and, based on studies on microorganisms, only a doubtful mutagenic activity. Information on teratogenic effects is contradictory. While some observations in man and a few animal studies suggest that lithium in concentrations in the order of those given to patients may cause malformations, other observations do not support this claim and the risk with a carefully controlled therapy is probably small. Until more information becomes available from ongoing lithium data registries, it is probably prudent to exercise caution in treating pregnant women with lithium during the period of cardiac organogenesis. No information is available on cancer caused by treatment with lithium, and it is highly unlikely that lithium is carcinogenic. PMID- 7491124 TI - Genotoxic potential of beta-carbolines: a review. AB - The mutagenic and co-mutagenic properties of harman, norharman and of some of their pharmacologically important derivatives are reviewed. These compounds do not behave as true mutagens, but rather interact, directly or indirectly with DNA, leading to various consequences. This unusual behaviour is most probably related to the particular structure of the chemical nucleus common to all beta carbolines which confers to the different derivatives the property to interact with various macromolecules and enzymatic systems. These interactions are compiled and discussed in this review. The alterations, by beta-carbolines, of some important enzymatic systems, e.g. cytochrome P-450, have been clearly demonstrated, yet many discrepancies and contradictions exist so that an interpretation of the results and the definition of some common mechanism appears premature. Since beta-carbolines are widely distributed in tissues and since they may modify and increase genotoxic and toxic consequences of other compounds, these interactions need to be clarified. PMID- 7491126 TI - Quantification of low-level radiation exposure by conventional chromosome aberration analysis. AB - Chromosome dosimetry, in its conventional form largely based on scoring of dicentrics and ring chromosomes in human blood lymphocytes, is the most widely distributed and reliable biological technique in radiological protection to estimate individual whole-body doses of about 100 mGy of low-LET radiation. Attempts to detect and quantify effects even of lower acute doses or protracted and chronic exposures have been repeatedly performed and the results revealed inherent limitations of this approach. Most relevant items, such as extrapolating from high-dose to low-dose effects, the influence of background frequency of dicentrics on the dose estimates, dose accumulation and concomitant temporal decline of the yields of unstable dicentrics or the statistical analyses of the data and their implications for quantifying low-level radiation exposure will be discussed in this report. PMID- 7491125 TI - Important biological variables that can influence the degree of chemical-induced aneuploidy in mammalian oocyte and zygotes. AB - The ability of certain chemicals to increase the frequency of aneuploidy in mammalian oocytes elicits concern about human health and well-being. This concernment exists because aneuploidy is the most prevalent class of human genetic disorders, and very little information exists about the etiology of aneuploidy. Although there are experimental models for studying aneuploidy in female germ cells and zygotes, these models are still being validated because insufficient information exists about the biological variables that can influence the degree of chemical-induced aneuploidy. In this regard, variables such as dose, solvent, use of gonadotrophins, mode and preovulatory time of chemical administration, time of cell harvest relative to the possibility of chemical induced meiotic delay, criteria for cytogenetic analysis and data reporting, and an introduction to differences between cell types and sexes are presented. Besides these variables, additional information is needed about the various molecular mechanisms associated with oocyte meiotic maturation and the genesis of aneuploidy. Also, differences between the results from selected chromosome analysis and DNA-hybridization studies are presented. Based upon the various biologic endpoints measured and the differences in cellular physiology and biochemical pathways, agreement among the results from different aneuploidy assays cannot necessarily be expected. To gain further insight into the etiology of aneuploidy in female germ cells, information is needed about the chemical interactions between endogenous and exogenous compounds and those involved with oocyte meiotic maturation. PMID- 7491128 TI - Comparison of autoradiography, liquid scintillation counting and immunoenzymatic staining of 5-bromo-2'-deoxyuridine for measurement of unscheduled DNA synthesis and replicative DNA synthesis in rat liver. AB - Different methods for evaluating unscheduled DNA synthesis (UDS) and replicative DNA synthesis (RDS) were studied in hepatocytes of F344 rats exposed in vivo to dimethylnitrosamine (DMN) or CCl4. Hepatocytes were isolated and incubated in Williams' medium E supplemented with either [3H]thymidine for autoradiography or 5-bromo-2'-deoxyuridine for immunoenzymatic staining. In the method of liquid scintillation counting, the cells were incubated with [3H]thymidine with or without hydroxyurea. The nuclear fraction was isolated and the incorporation of [3H]thymidine into nuclear DNA was determined by a liquid scintillation counter. DMN at doses of 0.625-5 mg/kg body weight induced UDS of 1.6-37.9 (0 dose; -6.9) net grains/nucleus measured by autoradiography and 337-1377 (0 dose; 177) dpm/microgram DNA in the presence of hydroxyurea measured by a liquid scintillation counter. CCl4 at doses of 50-400 mg/kg body weight induced RDS in 1.5-12.1% (0 dose; 0.12%) and 1.8-14.6% (0 dose; 0.16%) of cells with the methods of autoradiography and immunoenzymatic staining, respectively, and of 2991-24256 (0 dose; 324) dpm/microgram DNA in the absence of hydroxyurea with the method of liquid scintillation counting. Similar dose-dependent induction of UDS and RDS was observed with these methods. These results suggest that the methods of liquid scintillation counting and immunoenzymatic staining have almost the same sensitivity for measuring UDS and RDS as that of autoradiography. PMID- 7491127 TI - In-vivo genotoxicity of the alkaloid drug pilocarpine nitrate in bone marrow cells and male germ cells of mice. AB - Pilocarpine nitrate, an alkaloid drug of plant origin induces spindle disfunction in bone marrow cells of mice. Further studies were carried out to investigate its mutagenic effects in somatic and germ cells of mice by assessing chromosome aberrations at mitotic metaphase and as micronuclei in bone marrow cells and sperm-shape abnormality in cauda epididymides. The dose and time yield effects of the drug were investigated. The statistically significant results that were obtained for both chromosomal aberrations and micronucleus test but not for the sperm-shape abnormality test, indicated the genotoxicity of this compound in somatic cells but not in germ cells. PMID- 7491129 TI - DNA adducts induced by lipids and lipid peroxidation products: possible relationships to I-compounds. AB - A methanol-extractable lipid fraction of oats has been found previously to induce three specific I-compounds (age-dependent covalent DNA modifications) in female rat liver DNA, as detected by the 32P-postlabeling assay. The current report used an in vitro system to explore the possible mechanisms involved in the formation of these DNA derivatives. Ground oats or commercial oatmeal were extracted with methanol, and the extracts were incubated with rat lung DNA in vitro. DNA was recovered and analyzed by the nuclease P1-enhanced version of the 32P postlabeling assay. A number of adducts were induced by the in vitro reaction but none of them was identical by chromatographic analysis to oats-specific I compounds detected in vivo. Addition of rat liver microsomes and cofactors (NADPH or cumene hydroperoxide) to the in vitro reaction also failed to induce any of the oats-specific I-compounds. Pretreatment of oat lipids with soybean lipoxidase and oxygen enhanced formation of most adducts formed in vitro in a dose- and time dependent manner. Several of these adducts were related to peroxide derivatives of linoleic acid. Chromatographic evidence suggests that one of the major adducts is derived from 4-hydroxynonenal, a reactive intermediate lipid peroxidation product. This adduct was detectable in liver and kidney DNA of untreated rats and its level increased with age. These results were in line with previous in vivo results, suggesting that the oats-specific I-compounds are presumably formed via an indirect mechanism rather than by direct binding of oats components to DNA. PMID- 7491130 TI - Temporary variations in chromosomal aberrations in a group of agricultural workers exposed to pesticides. AB - The induction of chromosomal aberrations (CA) was studied in the peripheral lymphocytes of 29 male agricultural workers occupationally exposed to several pesticides. To investigate possible exposure-related changes in the frequency of CA, a longitudinal study has been conducted. Two blood samples were taken from each individual: one in a period of high exposure (spring-summer) and the other in a period of lower exposure (autumn-winter). Simultaneously, two matched control groups constituted by 29 and 24 healthy men, without indication of exposure to pesticides, were analysed. During the period of major exposure, the group of agricultural workers showed a significant increase in the frequency of CA, mainly of chromatid-type, when compared to the unexposed control group; nevertheless, this increase in the expression of CA was not found in the period of minor exposure. This finding could indicate that the frequency of CA is related to the intensity of the pesticide exposure, and that CA have a relatively short-life, recovering the control value a few months later. In addition to the cytogenetic analysis, biochemical and haemotological blood parameters were also analysed and no significant variations were detected. PMID- 7491132 TI - Growth rates of hprt and tk mutant CHO cell lines. AB - The growth rates of 31 X-ray-induced hypoxanthine phosphoribosyl transferase (hprt) deficient mutants of CHO-K1 cells were measured. Mutants had been classified as (1) full-deletion, (2) partial deletion or rearrangement, or (3) unchanged by Southern blot analyses. No relationship between growth rate and deletion type was observed. Even where all hprt-specific bands were missing, proliferation rates in culture were normal. Additionally, in CHO-AT3-2 cells, which are heterozygous at the tk locus, no difference in growth rates between a spontaneous hprt mutant and its parent was observed, although double hprt-tk-/- mutants grew more slowly. PMID- 7491131 TI - A new anthracycline with potent antileukemic activity exhibits reduced mutagenicity. AB - The mutagenicity of a new anthracycline (moflomycin) with potent antileukemic activity was studied by the Ames test in four strains of Salmonella typhimurium (TA97a, TA98, TA100 and TA102), and compared to the mutagenicity of doxorubicin, widely used as antineoplastic agent. Unlike doxorubicin, moflomycin displayed no mutagenic activity in strains TA98 and TA100. Low mutagenicity was only observed in TA102 strain and was not enhanced after metabolic activation. This result indicates that moflomycin induce mutagenicity by reverting base-pair substitution. The structural changes in the sugar moiety may be involved in the reduced mutagenicity of moflomycin. The low mutagenicity of moflomycin shown in this study enhances the potential advantage of this new derivative which displays a high antileukemic activity. PMID- 7491133 TI - The genotoxic potential of nicotine and its major metabolites. AB - Nicotine is a naturally occurring alkaloid found primarily in members of the solanaceous plant family, which includes tobacco. Nicotine is rapidly absorbed by humans and then metabolized, primarily by cytochrome P450's. Studies on the genotoxic potential of these metabolites are limited. Nicotine and four of its major metabolites: cotinine, nicotine-N'-oxide, cotinine-N-oxide, and trans-3' hydroxycotinine were evaluated for genotoxic potential in the Salmonella mutagenicity assay (strains TA98, TA100, TA1535, TA1537, and TA1538) at concentrations ranging from 0 to 1000 micrograms/plate and in the Chinese hamster ovary sister-chromatid exchange (SCE) assay at concentrations ranging from 0 to 1000 micrograms/ml. All assays were conducted with and without S9 metabolic activation. None of the five compounds increased the frequency of mutations or the frequency of SCEs. These results indicate that nicotine and its major metabolites are not genotoxic in the assays conducted. PMID- 7491134 TI - Estimated risk of transmission of the human immunodeficiency virus by screened blood in the United States. AB - BACKGROUND: In the United States, transmission of the human immunodeficiency virus (HIV) by blood transfusion occurs almost exclusively when a recently infected blood donor is infectious but before antibodies to HIV become detectable (during the "window period"). We estimated the risk of HIV transmission caused by transfusion on the basis of the window period associated with the use of current, sensitive enzyme immunosorbent assays and recent data on HIV incidence among blood donors. METHODS: We analyzed demographic and laboratory data on more than 4.1 million blood donations obtained in 1992 and 1993 in 19 regions served by the American National Red Cross, as well as the results of HIV-antibody tests of 4.9 million donations obtained in an additional 23 regions. RESULTS: We estimated that, in the 19 study regions, 1 donation in every 360,000 (95 percent confidence interval, 210,000 to 1,140,000) was made during the window period. In addition, it is estimated that 1 in 2,600,000 donations was HIV-seropositive but was not identified as such because of an error in the laboratory. We estimated that 15 to 42 percent of window-period donations were discarded because they were seropositive on laboratory tests other than the HIV-antibody test. When these results were extrapolated to include the additional 23 Red Cross service regions, there was a risk of one case of HIV transmission for every 450,000 to 660,000 donations of screened blood. If the Red Cross centers are assumed to be representative of all U.S. blood centers, among the 12 million donations collected nationally each year an estimated 18 to 27 infectious donations are available for transfusion. CONCLUSIONS: The estimated risk of transmitting HIV by the transfusion of screened blood is very small and nearly half that estimated previously, primarily because the sensitivity of enzyme immunosorbent assays has been improved. PMID- 7491135 TI - Brain glucose uptake and unawareness of hypoglycemia in patients with insulin dependent diabetes mellitus. AB - BACKGROUND: In patients with insulin-dependent diabetes mellitus (IDDM) whose treatment results in nearly normal mean plasma glucose concentrations, an unawareness of hypoglycemia can develop, and such patients are at increased risk for seizures and coma. We tested the hypothesis that during hypoglycemia, these patients would have normal glucose uptake in the brain and that consequently no sympathoadrenal activation would begin, resulting in an unawareness of hypoglycemia. METHODS: We measured glucose uptake in the brain at plasma glucose concentrations of 105 and 54 mg per deciliter (5.8 and 3.0 mmol per liter) in 24 patients with IDDM, stratified into three groups according to their glycosylated hemoglobin values (mean [+/- SD] values, 7.2 +/- 0.5, 8.5 +/- 0.4, and 10.2 +/- 1.3 percent) and compared the values for brain glucose uptake with those measured in 15 normal subjects at plasma glucose concentrations of 85 and 55 mg per deciliter (4.2 and 3.1 mmol per liter). We also recorded the subjects' hypoglycemic-symptom scores and measured their plasma concentrations of counterregulatory hormones. RESULTS: There was no significant change in the uptake of glucose in the brain (calculated as the uptake during hypoglycemia minus the uptake during normoglycemia) among the patients with IDDM who had the lowest glycosylated hemoglobin values (+0.6 +/- 2.0 mg [3.3 +/- 11.1 mumol] per 100 g of brain tissue per minute, P = 0.39). Conversely, glucose uptake in the brain fell in both the group with intermediate values (a decrease of 1.3 +/- 1.0 mg [7.2 +/- 5.6 mumol] per 100 g per minute, P = 0.009) and the group with the highest values (a decrease of 1.8 +/- 1.6 mg [10.0 +/- 9.0 mumol] per 100 g per minute, P = 0.01), as it did in the normal subjects (a decrease of 1.6 +/- 1.8 mg [9.0 +/- 10.1 mumol] per 100 g per minute, P = 0.003). The responses of plasma epinephrine and pancreatic polypeptide and the frequency of symptoms of hypoglycemia were lowest in the group with the lowest glycosylated hemoglobin values. CONCLUSIONS: During hypoglycemia, patients with IDDM who have nearly normal glycosylated hemoglobin values have normal glucose uptake in the brain, which preserves cerebral metabolism, reduces the responses of counterregulatory hormones, and causes an unawareness of hypoglycemia. PMID- 7491136 TI - Reduced incidence of preterm delivery with metronidazole and erythromycin in women with bacterial vaginosis. AB - BACKGROUND: Pregnant women with bacterial vaginosis may be at increased risk for preterm delivery. We investigated whether treatment with metronidazole and erythromycin during the second trimester would lower the incidence of delivery before 37 weeks' gestation. METHODS: In 624 pregnant women at risk for delivering prematurely, vaginal and cervical cultures and other laboratory tests for bacterial vaginosis were performed at a mean of 22.9 weeks' gestation. We then performed a 2:1 double-blind randomization to treatment with metronidazole and erythromycin (433 women) or placebo (191 women). After treatment, the vaginal and cervical tests were repeated and a second course of treatment was given to women who had bacterial vaginosis at that time (a mean of 27.6 weeks' gestation). RESULTS: A total of 178 women (29 percent) delivered infants at less than 37 weeks' gestation. Eight women were lost to follow-up. In the remaining population, 110 of the 426 women assigned to metronidazole and erythromycin (26 percent) delivered prematurely, as compared with 68 of the 190 assigned to placebo (36 percent, P = 0.01). However, the association between the study treatment and lower rates of prematurity was observed only among the 258 women who had bacterial vaginosis (rate of preterm delivery, 31 percent with treatment vs. 49 percent with placebo; P = 0.006). Of the 358 women who did not have bacterial vaginosis when initially examined, 22 percent of those assigned to metronidazole and erythromycin and 25 percent of those assigned to placebo delivered prematurely (P = 0.55). The lower rate of preterm delivery among the women with bacterial vaginosis who were assigned to the study treatment was observed both in women at risk because of previous preterm delivery (preterm delivery in the treatment group, 39 percent; and in the placebo group, 57 percent; P = 0.02) and in women who weighed less than 50 kg before pregnancy (preterm delivery in the treatment group, 14 percent; and in the placebo group, 33 percent; P = 0.04). CONCLUSIONS: Treatment with metronidazole and erythromycin reduced rates of premature delivery in women with bacterial vaginosis and an increased risk for preterm delivery. PMID- 7491137 TI - Association between bacterial vaginosis and preterm delivery of a low-birth weight infant. The Vaginal Infections and Prematurity Study Group. AB - BACKGROUND: Bacterial vaginosis is believed to be a risk factor for preterm delivery. We undertook a study of the association between bacterial vaginosis and the preterm delivery of infants with low birth weight after accounting for other known risk factors. METHODS: In this cohort study, we enrolled 10,397 pregnant women from seven medical centers who had no known medical risk factors for preterm delivery. At 23 to 26 weeks' gestation, bacterial vaginosis was determined to be present or absent on the basis of the vaginal pH and the results of Gram's staining. The principal outcome variable was the delivery at less than 37 weeks' gestation of an infant with a birth weight below 2500 g. RESULTS: Bacterial vaginosis was detected in 16 percent of the 10,397 women. The women with bacterial vaginosis were more likely to be unmarried, to be black, to have low incomes, and to have previously delivered low-birth-weight infants. In a multivariate analysis, the presence of bacterial vaginosis was related to preterm delivery of a low-birth-weight infant (odds ratio, 1.4; 95 percent confidence interval, 1.1 to 1.8). Other risk factors that were significantly associated with such a delivery in this population were the previous delivery of a low-birth weight infant (odds ratio, 6.2; 95 percent confidence interval, 4.6 to 8.4), the loss of an earlier pregnancy (odds ratio, 1.7; 1.3 to 2.2), primigravidity (odds ratio, 1.6; 1.1 to 1.9), smoking (odds ratio, 1.4; 1.1 to 1.7); and black race (odds ratio, 1.4; 1.1 to 1.7). Among women with bacterial vaginosis, the highest risk of preterm delivery of a low-birth-weight infant was found among those with both vaginal bacteroides and Mycoplasma hominis (odds ratio, 2.1; 95 percent confidence interval, 1.5 to 3.0). CONCLUSIONS: Bacterial vaginosis was associated with the preterm delivery of low-birth-weight infants independently of other recognized risk factors. PMID- 7491138 TI - Images in clinical medicine. Herpes simplex virus corneal dendrite. PMID- 7491139 TI - Familial aggregation of low birth weight among whites and blacks in the United States. AB - BACKGROUND: Studies have shown that the birth weight of infants is correlated with the birth weights of their siblings and their mothers. We investigated whether the birth weights of mothers and index children were jointly associated with the risk of low birth weight in the siblings of the index children. METHODS: We used data on the live-birth cohort of the 1988 National Maternal and Infant Health Survey. The analysis included 1691 white and 1461 black mothers, each of whom had two or more live-born, singleton children. Multiple logistic regression with generalized-estimation equations was used to assess the risk of low birth weight among an index child's siblings. Four groups were studied: that in which neither the mother nor the index child had low birth weight (group 1), that in which only the mother had low birth weight (group 2), that in which only the index child had low birth weight (group 3), and that in which both the mother and the index child had low birth weight (group 4). There was adjustment for other maternal and infant covariates. RESULTS: Among white siblings in groups 1, 2, 3, and 4, 3.6, 8.3, 21.2, and 38.9 percent, respectively, had low birth weight, as compared with 8.0, 19.0, 31.1, and 57.1 percent of black siblings. When group 1 was used as the reference group, the adjusted odds ratios (and 95 percent confidence intervals) for low birth weight in groups 2, 3, and 4 were 2.5 (1.4 to 4.3), 6.8 (4.7 to 9.8), and 15.4 (9.2 to 25.5), respectively, among white siblings and 2.6 (1.8 to 3.8), 4.7 (3.5 to 6.4), and 13.9 (9.2 to 20.9) among black siblings. These associations were consistently found for birth weights below 1500 g and those ranging from 1500 to 2499 g in both races and after stratification for the mother's age, parity, education, cigarette-smoking status, and weight and height before pregnancy and the infant's sex. CONCLUSIONS: Although selection and recall biases cannot be excluded with certainty, our data suggest a strong familial aggregation of low birth weight among both whites and blacks in the United States. PMID- 7491140 TI - Preoperative assessment of patients with known or suspected coronary disease. PMID- 7491141 TI - Seminars in Medicine of the Beth Israel Hospital, Boston. Clinical applications of research on angiogenesis. PMID- 7491142 TI - Case records of the Massachusetts General Hospital. Weekly clinicopathological exercises. Case 40-1995. A 71-year-old woman with right-lower quadrant pain and a cecal mass. PMID- 7491143 TI - Unawareness of hypoglycemia. PMID- 7491146 TI - Assessing the proposed Medicare reforms. PMID- 7491145 TI - The score on Medicare reform--minus the hype and hyperbole. PMID- 7491144 TI - Preterm delivery and low birth weight--a dire legacy. PMID- 7491147 TI - The oversupply of specialists and graduates of foreign medical schools. PMID- 7491148 TI - The oversupply of specialists and graduates of foreign medical schools. PMID- 7491149 TI - The oversupply of specialists and graduates of foreign medical schools. PMID- 7491150 TI - The oversupply of specialists and graduates of foreign medical schools. PMID- 7491152 TI - The oversupply of specialists and graduates of foreign medical schools. PMID- 7491151 TI - The oversupply of specialists and graduates of foreign medical schools. PMID- 7491153 TI - Early treatment of HIV infection. PMID- 7491154 TI - Early treatment of HIV infection. PMID- 7491155 TI - Early treatment of HIV infection. PMID- 7491156 TI - The long-QT syndrome. PMID- 7491157 TI - Sudden death during sports activities. PMID- 7491158 TI - Sudden death during sports activities. PMID- 7491159 TI - Sudden death during sports activities. PMID- 7491161 TI - Diverticulitis. PMID- 7491162 TI - Diverticulitis. PMID- 7491160 TI - Diverticulitis. PMID- 7491163 TI - Headphone neuralgia. PMID- 7491165 TI - Maternal eating patterns and birth weight of Mexican American infants. AB - Eating patterns of 549 Mexican American mothers were identified using dietary data from the United States Hispanic Health and Nutrition Examination Survey. These eating patterns were then used to investigate the relationship between maternal diet and infant birth weight. Principle components factor analysis was used to determine the structure of the maternal eating patterns. Seven distinct eating patterns were identified: nutrient dense, traditional, transitional, nutrient dilute, protein rich, high fat dairy, and mixed dishes. Stepwise multiple regression analysis was used to identify those eating patterns associated with birth weight. In addition to eating patterns, regression variables included body mass index, hemoglobin, gestational age at delivery, maternal age, infant gender, acculturation, marital status, income, education, and smoking during pregnancy. Regression results indicated that the nutrient dense (fruits, vegetables, low fat dairy, etc.) and protein rich (low fat meats, processed meats, and dairy desserts, etc.) eating patterns were associated with increased birth weight and that the transitional eating pattern (fats and oils, breads and cereals, high fat meats, sugar, etc.) was associated with decreased birth weight. Study findings suggest that the eating pattern methodology may be an appropriate tool for analyzing food frequency data in the investigation of diet and health relationships and for targeting dietary interventions. PMID- 7491164 TI - The adverse effects of tobacco smoking on reproduction and health: a review from the literature. AB - This paper compromises a short literary review of the adverse consequences of tobacco smoking on reproduction, as well as on infant and adult health. Furthermore, attention is drawn to the bad effects of smoking on the nutritional status. PMID- 7491167 TI - Taking the cooking out of food: nutrition & the national curriculum. AB - This is a discussion paper which generates some important research questions for educationalists. The teaching of food skills are at danger of being lost from the school cirriculum by changes imposed by the National Curriculum. This sinister (i.e., unexplained/undefined) development generates potential problems for the teaching of cookery and for the food and eating traditions of British society. The central concern for nutrition educationalists is that cooking is becoming more and more de-domesticated and consequently will become more systemised, more mass produced; in which case, young people need not be educated in basic cooking skills. PMID- 7491168 TI - Contribution of street foods to nutrient intakes by Nigerian adolescents. AB - The food and nutrient intake and the proportion of these supplied by street foods was studied in a group of 142 adolescent Nigerians, using structured questionnaire, 24-hour recall and weighted intake protocols. The results show that all the major groups of foods are represented in the menu and street food providing, overall, the major sources of dairy, meat, fish and egg. Mean intake of energy was 10.85 MJ for the cohort with street foods contributing almost 25%. For male subjects, 21% of the energy came from street foods while for females, the proportion was 29%. Also out of the mean intake of 62g total protein, street foods supplied over 50% for both male and female. The proportion was even higher for calcium (64%) and vitamin A (almost 60%). For all the other minerals and vitamins examined, street foods supplied over 50% of the intake. PMID- 7491166 TI - Old, poor & malnourished. AB - OBJECTIVE: To monitor the effects of low, fixed incomes on the dietaries and nutritional well-being of elderly people. METHODS: A questionnaire gathered data from a 'high income' sample (HIS) and a 'low income' sample (LIS) on income, eating and shopping patterns and nutritional awareness. A dietary diary provided the data required for a nutritional analysis of the foodstuffs consumed by the sample and inadequacies were identified by comparing nutrient intake to the Dept. of Health's (1991) Dietary Recommended Values (DRVs). RESULTS: The dietaries of the low income sample showed substantial shortfalls from those officially recommended and these are presented as the result, primarily, of income poverty and not the consequence of ignorance or mental decline. The older poor know what healthy eating entails; in most cases, they simply cannot afford and indeed, the dietaries are often inferior to the 'general poor', because of the idiosyncracies of their nutritional requirements. CONCLUSIONS: The inability of the older poor to purchase a healthy dietary cannot be solved merely by health education or budgeting skills. Primarily, they need more money. PMID- 7491170 TI - Note: vegetable oils. PMID- 7491169 TI - Note: schizophrenia--a mycotoxic element? PMID- 7491171 TI - Sugar has become the opium of the people. PMID- 7491173 TI - Nuclear medicine training in Europe. PMID- 7491172 TI - The future of nuclear medicine. PMID- 7491174 TI - Superiority of SPET to planar imaging in the detection of colorectal carcinomas with 111In monoclonal antibodies. AB - The aim of this study was to assess the contribution of single photon emission tomography (SPET) to planar imaging of colorectal carcinoma in patients being evaluated with 111In-labelled monoclonal antibodies CYT-103 (OncoScint CR/OV) or IVP ZCE 025. Planar and SPET scans from 110 colorectal carcinoma patients were scored individually as follows: 1=negative, 2=equivocal, 3=positive. The planar and SPET images identified 67 and 93 of 113 documented lesions, respectively. The planar and SPET findings were concordant in 55 patients. SPET converted planar findings from 1 to 3 in 11 patients and from 2 to 3 in 21 patients. SPET provided a better definition of the extent of the tumour in 21 patients. Both imaging tests were true-negative in five patients, and failed to detect tumours in six patients. We strongly recommended SPET in all patients undergoing immunoscintigraphy, since it identified tumours missed on planar scans in 35% of patients and provided additional information regarding tumour burden in 23% of patients. PMID- 7491175 TI - Thallium-201 myocardial SPET in strictly defined syndrome X. AB - We reviewed the exercise thallium-201 (201Tl) myocardial single photon emission tomography (SPET) results and clinical data of 28 patients, adhering to a strict definition of syndrome X. All of the patients had a normal resting left ventricular ejection fraction (LVEF) and no cardiac abnormalities. The results of the exercise electrocardiography (ECG) studies were evaluated and compared with the results of 201Tl SPET. Of the 28 patients, only 3 (11%) had a normal 201Tl SPET scan and 25 (89%) had an abnormal 201Tl SPET scan. We found that a normal 201Tl SPET scan was rare when adhering to a strict definition of syndrome X, and the results of exercise ECG did not correlate well with the perfusion defects observed on the 201Tl SPET scan. PMID- 7491176 TI - Comparison of myocardial ischaemia in post-infarction patients with and without angina: assessment by dipyridamole 201T1 imaging. AB - Ninety patients with old myocardial infarction were studied to evaluate the efficacy of intravenous dipyridamole 201Tl imaging for the detection of myocardial ischaemia in post-infarction patients, and to compare the prevalence of ischaemia in 63 patients with post-infarction angina (group I) and 27 patients without angina (group II). Thirty-four of the patients in group I and 15 of the patients in group II received coronary arteriography (CAG) for comparison; these were labelled groups IA and IIA, respectively. On 201Tl imaging, the incidence of scar with ischaemia in the infarct zone and scar with ischaemia at a distance were 72% and 42% in all patients, 89% and 52% in group I vs 33% and 19% in group II (P < 0.001 and P < 0.01, respectively). On CAG testing, the rates of infarct related recanalization vessel and multi-vessel disease were 76% and 68% in group IA vs 40% and 40% in group IIA (P < 0.05 and P > 0.05, respectively). Thus, dipyridamole 201Tl imaging is a useful modality and post-infarction angina a proper indicator in the detection of myocardial ischaemia in post-infarction patients PMID- 7491177 TI - Lung uptake of 201Tl in myocardial stress imaging: correlation with echocardiographic and scintigraphic variables of myocardial ischaemia. AB - The lung uptake of thallium-201 (201Tl) in planar imaging was evaluated in 384 patients enrolled in the SIRT study (Italian Multicentre Study on Thallium Reinjection). All patients underwent treadmill exercise, and at peak exercise 74 MBq of 201TI were injected. The patients were then evaluated after reinjection of 201Tl under baseline conditions. The left ventricle was subdivided into 16 echocardiographic and 15 scintigraphic segments. The best correlates of visually scored 201Tl lung uptake, of the lung heart ratio and lung wash-out were checked. The independent variables entered in the stepwise multiple-regression analysis were: the echocardiographic wall motion index, the scintigraphic perfusion score index, and the number of segments that improved by at least 1 and 2 grades after redistribution and after reinjection. We concluded that the lung:heart ratio and, in particular, lung wash-out do not correlate more closely than the visual score with the variables taken into consideration. This suggests that visually scored lung uptake of 201Tl is still of value in nuclear cardiology practice. The scintigraphic variables associated with reinjection do not seem to offer any additional information with respect to redistribution variables. We also analysed the impact on 201Tl lung uptake of several other index variables. Student's t test revealed that no single variable significantly alters lung uptake after stress testing. PMID- 7491179 TI - Comparative study of three automatic programs of left ventricular ejection fraction evaluation. AB - The aim of this study was to compare three automatic programs (P1, P2, P3) for evaluating radionuclide left ventricular ejection fraction (LVEF) and to emphasize the clinical consequences. Gated radionuclide ventriculography was performed in 73 subjects, 15 of whom were healthy and 58 of whom had experienced heart failure. All scintigraphic data were processed with the three programs. Good inter-observer, intra-observer and automatic-manual reproducibility were observed using each of the three programs. On the other hand, in the normal subjects, the three mean normal LVEF values were significantly different from each other (P1 = 77 +/- 5%, P2 = 63 +/- 7%, P3 = 68 +/- 8%; P < 0.0001) In the pathological patients, the values obtained with P2 were significantly different from those obtained using P1 and P3 (P1= 32 +/- 15%, P2 = 26 +/- 13%; P < 0.0001), Moreover, the linear regression studies between the three automatic programs were always significantly different from the identity line equation (y = x). This study shows that LVEF criteria for normality depend on the program used, and inter-program measurement of LVEF is poorly reproducible. Caution is recommended when comparing data obtained from different centres (or different computers), either in the follow-up of a given patient or in gathering results from patient groups. PMID- 7491178 TI - Whole-body 201Tl scintigraphy during one-leg exercise and at rest in normal subjects: estimation of regional blood flow changes. AB - To determine the effect of exercise intensity on regional blood flow in exercising and resting leg muscles and organs, we assessed relative changes in the regional distribution of 201Tl during one-leg ergometer exercise and at rest by whole-body scintigraphy as an estimate of regional blood flow changes in 10 healthy males. The regional 201Tl activity was expressed as a percentage of the whole-body activity, and relative changes in each region were compared to those at the anaerobic threshold (AT) and at peak exercise. In the legs, the distribution of 201Tl in the exercising thigh and calf increased significantly at the AT to 390.7 and 219.2% respectively of resting levels (both P < 0.01), but remained unchanged the resting thigh from rest to peak exercise. Conversely its distribution in the resting calf decreased significantly at the AT to 60.0% of the resting level (P < 0.01). However, there were no significant differences between AT and peak exercise. In the organs, the distribution of 201Tl in the lung, liver, spleen and kidneys decreased significantly at the AT to 88.4 +/- 10.4%, 67.0 +/- 11.5%, 62.4 +/- 11.8% and 67.0 +/- 8.6% respectively of resting levels (all P < 0.01). Redistribution of blood flow to the exercising leg occurs predominantly during mild to moderate exercise. Therefore, blood flow in the leg during strenuous exercise depends primarily upon an increase in cardiac output. The splanchnic organs are the major sources of redistribution. PMID- 7491180 TI - Validation of a dual-isotope technique using 123I-MIBG and 201Tl in the assessment of sympathetic reinnervation following heart transplantation: phantom and patient studies. AB - Dual-isotope studies using 123I and 201Tl allow the assessment of sympathetic reinnervation in patients following heart transplantation. 123I-meta-iodobenzyl guanidine (123I-MIBG) serves as a tracer of the integrity of the sympathetic nervous system. 201Tl is used for landmarking to allow better delineation of the myocardium due to faint 123I-MIBG accumulation in heart transplants. The aim of this study was to evaluate the influence of parameters such as crossover, attenuation and the 123I/201TI activity ratio on the assessment of the myocardial 123I-MIBG uptake ratio using phantom and patient studies. Crossover was calculated using the ratio: 201TI counts in the 123I energy window/201Tl counts in the 201Tl window. Phantom studies were performed using a rectangular phantom (RP) and a cardiac phantom (CP). The mean crossover from the 201Tl to the 123I energy window was 11.48% (RP) and 11.13% (CP). Depending on attenuation in water (depth of water 0-5 cm), crossover increased from 10.92 to 15.85% (RP) and from 11.05 to 15.79% (CP). In order to confirm that the experimental crossover results were equivalent to those obtained in patient studies, 15 patients underwent myocardial scintigraphy. After injection of 201Tl, a simultaneous dual-window acquisition was performed to assess crossover from the 201Tl to the 123I window. The mean crossover was 15.35%, as high as the crossover assessed in phantom studies, taking into account attenuation and scatter caused by the chest wall. In order to reduce 201Tl crossover, the 123I activities were six times as high as the 201Tl activities.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7491181 TI - Interpretation of lymphoscintigrams in suspected lymphoedema: contribution of delayed images. AB - Interpretation of lymphoscintigrams in patients with extremity oedema may be adversely affected by any variation in the acquisition protocol. The purpose of this study was to determine whether delayed (2-24 h post-injection) views provide information incremental to the first 60 min of acquisition. In a 5-year period, 31 patients (10 males, 21 females, mean age 46 years) with extremity oedema or recurrent cellulitis underwent lymphoscintigraphy using 99Tcm-antimony trisulphide colloid (99Tcm-Sb2S3). All subjects underwent an initial dynamic acquisition of the pelvis and legs for 60 min, with further images obtained after 2-24 h. Films and reports were retrospectively reviewed by an experienced observer to determine whether delayed views provided information incremental to early images. Using standard qualitative criteria, lymphoscintigrams were classified as normal (n = 12) or consistent with lymphoedema (n = 19). However, 6 of 19 (32%) patients with lymphoedema displayed normal transit of radiopharmaceutical and a normal appearance of regional lymph nodes on views acquired up to 1 h after injection; localized dermal backflow (n = 5) or lymphocoele (n = 1) were seen only on delayed images (P < 0.02). These data indicate that delayed lymphoscintigraphy images are important for the diagnosis of lymphoedema and should be obtained even when lymphatic transit of 99Tcm-Sb2S3 and lymph node appearance at 1 h are normal. PMID- 7491182 TI - Evaluation of thyroid nodules for malignancy using 99Tcm-sestamibi. AB - 99Tcm-sestamibi (99Tcm-MIBI) is used for myocardial perfusion imaging but has also been reported to localize in tumours. The usual thyroid scanning radionuclide is 99Tcm-pertechnetate. Altogether, 161 patients with clinically solitary thyroid nodules had both 99Tcm-MIBI and 99Tcm-pertechnetate thyroid scans, with the nodules being reported as cold, warm or hot. Fine-needle aspiration cytology (FNAC) and surgery were performed in those patients who consented to these procedures. Of 131 patients who had FNAC, only 58 proceeded to surgery. In the surgically treated group, 14 of 58 (24%) were confirmed to have thyroid cancer, whereas 44 of 58 (76%) had benign lesions. The 14 cancerous nodules were cold on the 99Tcm-pertechnetate scan, whereas with 99Tcm-MIBI 11 were warm and 3 were either cold or hot nodules. Of the 44 benign lesions, 18 were cold, 9 were warm and 17 were hot nodules. In those 131 patients who had FNAC, the cytology was reported as benign in 120 of the nodules and malignant in 11. The three false-negative cytologies were reported as follicular adenomas. The benign lesions noted on FNAC and surgery were thyroiditis, adenomas and haemorrhagic or colloid cysts. The results from the 58 surgically treated patients suggest that the warm nodules would need surgery, whereas the cold and hot nodules are unlikely to be malignant. The overall sensitivity of the 99Tcm MIBI scan was 79% and the specificity 80%, with the warm nodule on 99Tcm-MIBI scan having a positive predictive value of 55% and a negative predictive value of 92%. PMID- 7491183 TI - Positive thyroid cancer scintigraphy using 99Tcm-tetrofosmin (Myoview): a preliminary report. AB - 99Tcm-tetrofosmin imaging was performed in 23 thyroid cancer patients and compared with whole-body 131I scans, neck ultrasonography and plasma thyroglobulin levels. Of 22 patients with differentiated thyroid cancer, 8 showed identical images on 131I scan and tetrofosmin; 1 patient with medullary cancer had identical images with 99Tcm-DMSA and 99Tcm-tetrofosmin, the findings being confirmed by sonography. The potential usefulness of 99Tcm-tetrofosmin imaging in the follow-up of thyroid cancers is discussed. PMID- 7491185 TI - Cobalt-57 SPET in relapsing-progressive multiple sclerosis: a pilot study. AB - Multiple sclerosis (MS) is predominantly a progressive immune-mediated disease of the white matter in the brain. We used single photon emission tomography (SPET) and cobalt-57 (57Co) as a calcium (Ca) analogue to visualize brain tissue damage, based on the fact that Ca influx occurs in both cell death and T-lymphocyte activation in MS. The aim of this study was to determine if 57Co-SPET detects MS lesions and, if so, to compare this with clinical data on the patient. Five MS patients underwent neurological examination including Expanded Disability Status Scale (EDSS) assessment and 57Co-SPET, using a single-headed camera. All available data were compared. The lesions were recognized as areas of increased signal intensity, although the poor count rate did not allow any statistical quantification. A relationship between one 57Co-SPET parameter (cobalt plaque load) and EDSS was demonstrated. In conclusion, this pilot study suggests that 57Co-SPET using a single-headed camera is not an appropriate imaging modality in MS. To obtain a more favourable signal-to-noise ratio, the use of a multi-headed camera, the administration of a higher activity of 57Co and a longer acquisition time are recommended. Validation of this method among a larger group of patients and a comparison with healthy volunteers is needed. PMID- 7491184 TI - Oestrogen and bone density: a comparison of tamoxifen and hypo-oestrogenaemia. AB - The effect of the anti-oestrogen tamoxifen on bone density in post-menopausal women with stage II breast cancer (n = 35) was compared with two untreated hypo oestrogenaemic groups (premature ovarian failure, n = 20; prolactinoma, n = 19) and a group of women with prolactinomas and normal oestrogen levels (n = 12). Bone densities were normal in the euoestrogenaemic groups, but significantly reduced in both hypo-oestrogenaemic groups. Repeat measurements were available for 31 of the 35 tamoxifen-treated subjects after an interval of 33.9 +/- 14.7 months. The anti-oestrogen action of tamoxifen was not associated with accelerated bone loss of the spine or hip, an outcome that was clearly demonstrable in hypo-oestrogenaemic disorders. PMID- 7491187 TI - Preventing renal uptake of 111In from labelled monoclonal antibody fragments. PMID- 7491186 TI - Differential kinetics of parotid and submandibular gland function as demonstrated by scintigraphic means and its possible implications. AB - Is the differential kinetics of secretion of saliva between the parotid and submandibular glands responsible for the relative difference in radiosensitivity? Radiation-induced dysfunction is more often noted in the parotid glands than the submandibular glands. This difference in susceptibility is often attributed to the difference in the cellular composition of these glands. During our studies using 99Tcm-pertechnetate to quantitate salivary gland function in patients treated with radioiodine, we noted frequent unstimulated discharge of saliva from the submandibular glands compared with parotid secretion. We hypothesize that the relative resistance of the submandibular glands could be due to the differential kinetics of the parotid and submandibular glands, as evidenced by using 99Tcm pertechnetate to study their function. PMID- 7491188 TI - MR angiography in pediatric neurological disorders. AB - MR angiography using 3D and 2D time-of-flight techniques were used to evaluate pediatric neurological disorders. MRA (arteriography) and MRV (venography) were abnormal in 63 and 45 cases, respectively. Conventional cerebral angiography was performed in 30 cases. These techniques were compared with MRI and conventional angiography. In addition, the value of MR angiography for surgical planning was subjectively evaluated. Our results showed that intracranial vessels were invariably better shown on MR angiography than on MRI. MRA and MRV were most useful in evaluating vascular distortions related to congenital brain malformations and intracranial tumors. MRA was valuable in detecting arterial narrowing but overestimated the degree of stenosis compared with conventional angiography. MRV was the technique of choice for evaluation of dural sinus and cerebral venous thrombosis and compression. MRA played little to no role in preoperative planning of vascular malformations and aneurysms. It did not appear to be accurate in assessing tumor vascularity or lesions in small arteries and arteritis. PMID- 7491189 TI - Local vascular CO2 reactivity in the infant brain assessed by functional MRI. AB - The local cerebral vascular response to hyperventilation was investigated in five distressed, intubated infants by means of a T2 sensitive gradient-echo MRI technique at 1.5 T. In one preterm infant, the MR signal change during hyperventilation was sparse. In four term infants, the mean MR signal of the brain slice investigated decreased by 1.2-2.6% per kPa change in PCO2 as a reflection of decreased cerebral blood flow during hyperventilation. Pixel-wise analysis revealed absence of vascular response in the basal ganglia, the thalamus or in the occipital region. In two adult controls, who hyperventilated voluntarily, the vascular reactivity was homogeneously distributed predominantly over the grey matter. The experiments demonstrate that local impairment of vascular CO2 reactivity in the distressed infant brain can be detected by T2 sensitive gradient-echo MRI, which is also known as functional MRI. PMID- 7491190 TI - Optimal cost-effective timing of cranial ultrasound screening in low-birth-weight infants. AB - Our aim in this study was to determine whether delaying the initial screening cranial ultrasound on infants of low birth weight until the 2nd week of life affects clinical diagnosis and cost of patient care. Data were reviewed on all premature infants of less than 33 weeks gestation or less than 1500 g birth weight admitted to the Neonatal Intensive Care Unit between January 1989 and August 1992. Babies admitted before August 1991 were screened on day 4 or 5 with a second scan frequently performed on day 14; babies admitted after that date were screened once between days 10-14. Populations were compared for (1) proportion of ultrasound findings considered normal on final diagnosis; (2) incidence of major and minor abnormalities; (3) periventricular leukomalacia (PVL); (4) change in diagnosis from 1st to 2nd week; and (5) number of studies performed per patient. The study group was composed of 499 eligible infants. Demographic features of infants screened in the 1st vs. 2nd week of life were similar, with comparable diagnoses of major and minor intracranial hemorrhage and ventricular dilatation; however, more patients screened in the 1st week had questionable PVL diagnosed (p = 0.04). There was a significant decrease in the number of scans per patient in the group screened in the 2nd week (p < 0.004). Routine screening may be delayed until the 2nd week without compromising patient care. Widespread use of a similar screening protocol would result in significantly fewer studies being performed, with an estimated saving, in the USA, of more than $3 million annually. PMID- 7491191 TI - Atypical and equivocal sonographic features of the spinal cord in neonates. AB - Thanks to the wider use of sonography to examine the spinal content in the neonate, normal anatomy and anomalies may be detected easily. Yet, unusual sonographic patterns are also observed. These must be differentiated from true pathologies. During a prospective study of 103 neurologically asymptomatic neonates, atypical sonographic patterns were found in 16 patients, corresponding to normal variants in 13. Nine of these 13 patients presented with a widening of the distal part of the central echo complex (one had a dilated ventriculus terminalis). Nerve roots of the cauda equina were disposed asymmetrically in three patients; the spinal cord movements were still present. In two of these babies, this distribution was associated with thin arachnoid pseudocysts. One patient presented with transitorily hyperechoic and narrow subdural spaces, probably related to neonatal dehydration. None of the 13 patients showing normal variants required any treatment. The other 3 patients (of 16) presented with equivocal entities of unknown evolution: sonographic tethered cord, fibrolipoma of the filum terminale and epidural varices. Sonography is highly accurate in evaluating the spinal cord content and aids differentiation of normal and normal variants from equivocal or pathological entities. PMID- 7491192 TI - Ependymoblastoma: MR presentation. A case report and review of the literature. AB - Ependymoblastoma is a rare and devastating tumor of childhood considered by most to be a subtype of primitive neuroectodermal tumors. We present the first detailed MRI description of this entity with a review of the limited pathologic and radiologic literature concerning this tumor. PMID- 7491193 TI - Transtentorial herniation of the fourth ventricle. AB - Transtentorial herniation (TTH) of the fourth ventricle is the result of a progressive enlargement of an isolated fourth ventricle which herniates through the tentorial incisure into the middle cranial fossa. The characteristic CT-MR and neurosonographic findings are described. PMID- 7491194 TI - Unilateral cortical dysplasia associated with contralateral hyperplasia of the brainstem. AB - Magnetic resonance imaging of a patient with unilateral frontotemporal cortical dysplasia is described. The brainstem showed ipsilateral hypoplasia, a usual but subjective radiological finding reportedly associated with unilateral cortical dysplasias, which inherently may cause underestimation of contralateral brainstem changes. In this patient contralateral hyperplasia of the brainstem was also present, evidenced especially by apparent hyperplasia of the pyramid of the medulla oblongata, where the corticospinal tract runs. Such hyperplasia of the brain stem may reflect a compensation reaction in order to increase the number of fibers in the corticospinal tract within the normal hemisphere, which are actually deficient (hypoplastic) in the contralateral, diseased hemisphere. PMID- 7491195 TI - A vein of Galen aneurysm with an abnormal drain system: MRI findings. AB - The case of a premature girl with a vein of Galen aneurysm is reported. The lesion was detected by ultrasonography. MRI provided complete anatomical information, obviating the need for initial angiography for preoperative evaluation of the malformation. Venous drainage through a persistent embryonic falcine sinus and associated atresia of the straight sinus could be identified, while venous thrombosis and parenchymal damage could be excluded. PMID- 7491196 TI - Cerebral hemorrhage in alloimmune thrombocytopenia. AB - Intracranial hemorrhages are rare in term infants. We describe such an infant born with multiple foci of intraparenchymal hemorrhage in various stages of maturation, caused by maternal platelet-specific anti-PlA1 antibodies that were delivered transplacentally to the developing fetus. PMID- 7491197 TI - Oculoauriculovertebral spectrum: new manifestations. AB - We present a case of oculoauriculovertebral spectrum (hemifacial microsomia, Goldenhar's syndrome) found shortly after birth with right iliac hypoplasia, absent right pubic and ischial ossification centres and posterior urethral valves, which have not previously been described in this spectrum. PMID- 7491198 TI - Localized pneumothorax with lobar collapse and diffuse obstructive airway disease. AB - Localized pneumothorax adjacent to a collapsed lobe has been reported in children with bronchial obstruction. We present our findings in seven children with a similar phenomenon occurring in association with diffuse obstructive airway disease. The children, aged from 3 weeks to 17 years, were admitted for diffuse obstructive airway disease and, subsequently, developed lobar collapse with adjacent localized pneumothorax. In five of the seven patients there was a paradoxical shift of the mediastinum toward the side of the pneumothorax. In six cases, the pneumothorax resolved spontaneously with lobar reexpansion. A conservative treatment approach to patients with this constellation of radiographic findings appears justified. PMID- 7491199 TI - Radiographic findings after pectus excavatum repair. AB - Postoperative radiographic changes after pectus repair have not previously been described. We reviewed the chest radiographs of 65 children who underwent surgical repair of pectus excavatum between early 1985 and mid-1992. After operation, 64 children showed atelectasis, 38 had pleural effusions, 30 showed a congestion or edema pattern, 12 had pneumothorax, 21 showed fixation strut tilt or rotation, and 4 had pneumomediastinum. Despite the sometimes worrying radiographic appearance, these children uniformly did well clinically. PMID- 7491201 TI - Subclavian artery compression from a chest tube after thoracotomy in a premature infant. AB - A premature baby underwent thoracotomy for repair of esophageal atresia and tracheoesophageal fistula, during which time a right chest tube was placed. In the immediate postoperative period, right subclavian artery occlusion was recognized and relieved by partial withdrawal of the chest tube, which had migrated high into the apex of the right hemithorax. PMID- 7491200 TI - MR of right aortic arch with mirror-image branching and a left ligamentum arteriosum: an unusual cause of a vascular ring. AB - An unusual case is presented of a vascular ring due to a right arch with mirror image branching and a left ligamentum arteriosum from the descending aorta to the left pulmonary artery. There have been only four previous reports of this rare arch anomaly. In addition, the patient had no underlying congenital heart lesions, which is also very unusual in patients with right aortic arch with mirror-image branching. PMID- 7491202 TI - Bladder neck obstruction caused by a large simple ureterocele in a young male. AB - The most common clinical presentation of ureterocele is infection of the urinary tract. Much less common is obstruction of the bladder outlet by prolapse of the ureterocele into the urethra. Even less common is simple occlusion of the bladder outlet by a nonprolapsing ureterocele. We present such a case where a large, simple ureterocele in a young male led to such obstruction. PMID- 7491204 TI - A "flipped" kidney in utero in an infant with a double collecting system and a Gartner's duct cyst with a vaginal ectopic ureter. AB - We report an infant with two unique anatomic abnormalities. A "flipped" kidney in utero is described with the association of a Gartner's duct cyst and a vaginal ectopic ureter with a duplicated collecting system. PMID- 7491203 TI - Nephrocalcinosis in a child with autosomal dominant polycystic kidney disease and a prolapsing ectopic ureterocele. AB - Although autosomal dominant polycystic kidney disease commonly presents in adults, it can occur in children. Usually, renal calcification in patients with autosomal dominant polycystic kidney disease is manifested as calculi or as hemorrhage into a renal cyst. An ectopic ureterocele is a well-known finding in patients with renal duplication. To our knowledge, this is the first case report of a child who had combined findings of autosomal dominant polycystic kidney disease, nephrocalcinosis, and an obstructing ectopic ureterocele. PMID- 7491205 TI - Prenatal diagnosis of glomerulocystic kidney disease in short-rib polydactyly syndrome type II, Majewski type. AB - An 18-year-old gravida 1 para 0 female was presented at 35 weeks' gestation with severe oligohydramnios. Sonography revealed a fetus with hydrops, bilaterally enlarged echogenic kidneys, and short extremities. The infant died at birth and autopsy was compatible with short-rib polydactyly syndrome type II, Majewski type. Renal histology was consistent with glomerulocystic disease. PMID- 7491206 TI - Renal dysplasia in infants: appearance on 99mTc DMSA scintigraphy. AB - Infantile renal dysplasias, including multicystic dysplastic kidneys (MCDK), are reported rarely to accumulate radiopharmaceuticals on renal scintigraphy. 99mTc DMSA is a highly sensitive tracer for detecting functioning renal cortical tissue and may be more suited to studying renal dysplasia than 99mTc DTPA. We reviewed the ultrasound studies and 99mTc DMSA scintigrams of 42 infants (age range 1-12 months) with known or suspected MCDK. Overall, uptake on 99mTc DMSA scintigraphy was evident in 6/41 (15%) dysplastic kidneys. Of the 18 patients who underwent nephrectomy, histopathological examination revealed that uptake correlated closely with the presence of mature renal cortical tissue in the affected kidney. Our study shows that a small, but significant number of MCDK will show low-grade uptake on DMSA scintigraphy. This finding may be relevant given the reliance placed on renal scintigraphy in planning treatment for infants with suspected MCDK, particularly with the increasing trend for the non-operative management of this condition. PMID- 7491207 TI - Ureteropelvic junction obstruction in infants mimicking ureterovesical junction obstruction on sonography. AB - We present two infants with ureteropelvic junction obstruction with very large renal pelves, which extended to the level of the urinary bladder. In both cases, the distal aspect of the renal pelvis simulated a dilated distal ureter, leading to the incorrect sonographic diagnosis of ureterovesical junction obstruction. Awareness of this potential pitfall on sonography is important when imaging infants with prenatal diagnosis of hydronephrosis. PMID- 7491208 TI - Congenital intraluminal tracheal cyst causing an obstructed left lung in a newborn. AB - Intraluminal tracheal or bronchial cysts are rare. We present a congenital cyst of the distal trachea which caused obstruction of the left main-stem bronchus. The chest radiograph obtained at birth showed an expanded, irregularly opaque left lung. Within several hours, the opacity cleared and the lung became markedly hyperexpanded. The cyst ruptured spontaneously with resolution of the symptoms. PMID- 7491209 TI - Sonographic diagnosis of systemic venous gas in a patient with pneumatosis intestinalis. AB - The detection of systemic venous gas due to pneumatosis intestinalis in patients with portal hypertension has not been described. Since sonography is commonly used to investigate conditions associated with pneumatosis intestinalis, it is important to recognize their sonographic appearance and clinical implications. PMID- 7491210 TI - Hypertrophic osteoarthropathy caused by lipoid pneumonia. AB - A case of hypertrophic osteoarthropathy in a 5-year-old Saudi boy is presented. The child developed painful joints, was found to have clubbing of fingers and radiologically hypertrophic osteoarthropathy was demonstrated. The underlying cause of the condition was ingestion of animal fat in infancy, with a history of ghee oil aspiration which caused a chronic lung inflammation, lipoid pneumonia, proven by lung biopsy. PMID- 7491211 TI - Glutaric aciduria: improved MR appearance after aggressive therapy. AB - Advances in understanding the metabolic abnormalities which cause glutaric aciduria allow biochemical diagnosis on the basis of deficient enzyme and the potential for therapy. Brain abnormalities associated with this inborn error of metabolism have been demonstrated with CT and MR. The findings typically described are atrophy of the fronto-temporal regions with large insular cisterns and diffuse white matter hypodensities. We present a patient with glutaric aciduria, confirmed by enzymatic assay, who had these findings on CT and MR examination. Repeat imaging demonstrated significant improvement after dietary therapy and aggressive prevention of catabolism during febrile illness. PMID- 7491213 TI - Bilateral orbital granulocytic sarcoma (chloroma) preceding the blast phase of acute myelogenous leukemia: CT findings. AB - Granulocytic sarcoma (chloroma) is an uncommon malignant neoplasm associated with myelogenous leukemias. Its appearance may precede the clinical manifestations of the acute leukemic phase by months to years. We report the CT findings of a case of bilateral intraorbital granulocytic sarcoma which preceded the blast phase of acute myelogenous leukemia (AML) and led to its diagnosis. An awareness of granulocytic sarcoma and its CT appearance may expedite the diagnosis of AML or prompt close monitoring of those cases of granulocytic sarcoma antedating the appearance of AML. PMID- 7491212 TI - Resolution of a proximal humeral defect in type-1 Gaucher disease by enzyme replacement therapy. AB - Although rarely they are a normal variant in children, significant defects in the medial aspect of the proximal humeral metaphysis occur in patients with Gaucher disease due to cortical infiltration and erosion of the periosteum by Gaucher cells. Such changes may lead to pathological fractures in Gaucher patients. These crescentic erosions resolved in a 19-year-old patient with type-1 Gaucher disease following enzyme replacement therapy. PMID- 7491214 TI - Hyperechoic ependyma in the neonate. PMID- 7491215 TI - Congenital ventricular diverticulum. PMID- 7491216 TI - The impact of fetal screening on indications for cystourethrography in infants. PMID- 7491217 TI - Annual summary of vital statistics-1994. AB - Recent trends in the vital statistics of the United States continued in 1994, including decreases in the number of births, the birth rate, the age-adjusted death rate, and the infant mortality rate. Life expectancy increased slightly to 75.7 years. Only marriages reversed the recent trend with a slight increase in 1994. An estimated 3,979,000 infants were born during 1994, a decline of < 1% from 1993. The birth rate was 15.3 live births per 1000 population, a 1% decline. These decreasing rates reflect a decline in the fertility rate to 67.1 live births per 1000 women aged 15 to 44 years. Final figures for 1993 indicate that fertility rates declined for all racial groups, by 1% for white women (to 65.4) and 3% for black women (to 80.5). The fertility rate for Hispanic women (106.9) was 84% higher than that for non-Hispanic white women and 31% higher than for non Hispanic black women. Between 1991 and 1993, birth rates for teenage mothers remained virtually unchanged, and abortion rates have steadily declined, suggesting that teenage pregnancy rates are levelling off. The number and proportion of births to women over age 30, however, continued to rise. The rate of births to all unmarried women (45.3 per 1000 in 1993) has been stable for 3 years. Prenatal care utilization improved in 1993; 79% of women initiated care in the first trimester and < 5% had delayed care or no care. Improvements occurred among nearly all racial and ethnic groups. Reported smoking during pregnancy declined to 15.8% in 1993 from 16.9% in 1992. The proportion of babies delivered by cesarean section was 21.8% in 1993, a 2% decrease from 1992. Between 1992 and 1993, the rate of low birth weight (LBW) rose slightly to 7.2%, while very low birth weight (VLBW) remained stable at 1.3%. Most of the increase in LBW occurred among white infants and reflected, primarily, an increase in the proportion of multiple births. The black/white ratio in LBW continued to increase to more than two-fold with the largest difference recorded among term and postterm infants. Age-adjusted death rates in 1994 were lower for heart disease, malignant neoplasm, pulmonary diseases, other accidents, and homicides. The age-adjusted death rate for human immunodeficiency virus disease continued to rise to 15.1 in 1994. The infant mortality rate declined 4% in 1994, to 7.9 per 1000, the lowest rate ever recorded in the United States. The decline was primarily in neonatal mortality.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 7491218 TI - Antiphospholipid antibodies in pediatric systemic lupus erythematosus. AB - OBJECTIVE: Antiphospholipid antibodies (aPLs) have been extensively studied in adults with systemic lupus erythematosus (SLE) and have been associated with arterial and venous thrombosis, thrombocytopenia, neurologic disorders, and recurrent fetal loss. In contrast, very little is known about the frequency and clinical significance of aPLs in pediatric SLE. This study was designed to determine the frequency of aPLs in pediatric SLE and the temporally associated clinical manifestations. DESIGN: We studied 29 consecutive patients with onset of SLE in childhood seen in the Pediatric Rheumatology Clinic at the University of Pittsburgh, Children's Hospital, between 1985 and 1992. We defined aPL as the presence of a lupus anticoagulant (LAC), immunoglobulin G or immunoglobulin M anticardiolipin antibodies (aCLs), or a biologic false-positive serologic test for syphilis determined by a VDRL test. Clinical manifestations were temporally correlated to the presence of aPLs if they occurred within 6 months. RESULTS: Overall, 19 (65%) of 29 children with SLE had one of the three laboratory abnormalities defining aPL. LAC was detected in 16 (62%) of 26, aCL in 18 (66%) of 27, and false-positive VDRL test results in 11 (39%) of 28. Twenty-five of the 29 patients had all three tests performed. In 10 patients, all three tests were abnormal. The presence of thrombosis in 7 patients (4 venous, 2 arterial, and 1 both) was associated with a positive aPL, specifically aCL. The presence of an aPL was significantly associated with anti-double-stranded DNA antibodies, but not with neuropsychiatric manifestations or with thrombocytopenia. The presence of an aCL was significantly associated with hemolytic anemia. A prolonged prothrombin time, in the setting of an LAC (all with a prolonged activated partial thromboplastin time), was associated with life-threatening disease in 6 of 15 patients. CONCLUSION: Sixty-five percent of 29 consecutive pediatric patients with SLE had evidence of aPL. The presence of aPL, specifically aCL, was significantly associated with thrombotic events. The presence of a prolonged prothrombin time in the setting of an LAC may be a marker of more serious disease in pediatric SLE. PMID- 7491219 TI - Partial deficiency of surfactant protein B in an infant with chronic lung disease. AB - OBJECTIVE: To evaluate components of pulmonary surfactant and identify mutations in the surfactant protein B gene (SP-B) of a term infant with severe respiratory distress and chronic lung disease. PATIENT AND TESTING: Respiratory distress developed in an infant delivered at term, and he required extracorporeal bypass support for 2 weeks. Until his unexpected death at 9.5 months, he was ventilator and oxygen dependent and required continual dexamethasone therapy. Tracheobronchial lavage samples were analyzed for content of surfactant proteins (SPs), and DNA from blood samples were sequenced and analyzed by polymerase chain reaction restriction analysis for the presence of SP-B gene mutations. Surfactant lipid composition and function, the contents of SPs and their messenger RNAs (mRNAs), and the immunostaining pattern for SPs were determined in postmortem lung tissue. RESULTS: The lavage sample contained SP-A but not SP-B, and DNA restriction analysis indicated that the patient and his mother were heterozygous for the previously described 121ins2 mutation of SP-B. Postmortem lung tissue contained normal levels of SP-A and its mRNA, a low but detectable level of SP-B, and near normal content of SP-B mRNA. SP-C was abundant on staining, and some 6 kd precursor was present in tissue. A surfactant fraction was deficient in phosphatidylglycerol and was not surface active. On DNA sequencing, a point mutation was found in exon 7 of the patient's SP-B gene allele without the 121ins2 mutation, resulting in a cysteine for arginine substitution, and the father was a carrier for the same mutation. CONCLUSIONS: We describe a patient who is a compound heterozygote with a new mutation and only a partial deficiency of SP-B. Some forms of inherited SP-B deficiency may have low expression of immunoreactive and possibly functional SP-B with milder lung disease and longer survival. These infants may benefit from glucocorticoid therapy and may not develop antibodies to SP-B after either lung transplant or gene therapy. PMID- 7491220 TI - Neurologic manifestations in children with Lyme disease. AB - OBJECTIVE: Lyme disease (LD) is a tick-borne spirochetal infection with a wide range of neurologic and non-neurologic manifestations. The clinical diversity of LD and limitations in serologic diagnosis often make it difficult to document the diagnosis of neuroborreliosis with certainty. METHODS: We reviewed clinical manifestations in 97 seropositive children with particular attention to neurologic manifestations. Diagnostic criteria used in other case surveys were applied to determine how often a definitive diagnosis of neuroborreliosis could be made in children. RESULTS: Of 69 children who met criteria for LD, 32% (22) had new neurologic signs, 73% (16) of which were accounted for by facial palsy and aseptic meningitis. Five of those with neurologic findings also had erythema migrans (EM), and one had both EM and arthritis. Among those with neurologic involvement, boys outnumbered girls two to one. Neurologic abnormalities resolved spontaneously in five children before their serologic results were known. CONCLUSION: In our series, only 27% of children with neurologic abnormalities due to LD had a history of EM or arthritis. Seropositivity commonly constituted the primary basis for diagnosis of LD. Despite its nonspecificity, seropositivity for LD in children with neurologic symptoms usually signifies active neuroborreliosis. PMID- 7491221 TI - The prevention of coronary artery aneurysm in Kawasaki disease: a meta-analysis on the efficacy of aspirin and immunoglobulin treatment. AB - OBJECTIVE: Varying observations have been made concerning the use of aspirin (ASA) and/or intravenous immunoglobulin (IVIG) in the prevention of coronary artery aneurysm (CAA) in children with Kawasaki disease. A meta-analysis of published articles on the subject was conducted to evaluate the reported efficacy of these therapies. METHODS: All published studies in all languages from 1967 through 1993 obtained from MEDLINE and EMBASE were considered, and a defined set of inclusion and exclusion criteria selected the studies for analysis. These studies were grouped based on whether the children in the studies received: (1) ASA alone, (2) low IVIG (< or = 1 g/kg) and ASA, (3) high IVIG (> 1 g/kg) and ASA, (4) single IVIG (> 1 g/kg) and ASA, (5) high IVIG and low ASA (< or = 80 mg/kg), or (6) high IVIG and high ASA (> 80 mg/kg). Studies that satisfied the test for homogeneity were subjected to further analysis. The best estimate of the true proportion of CAA as well as the 95% confidence interval for each group were calculated at 30 and 60 days. Hypothesis testing was conducted to determine the statistical significance of the calculated difference in each compared treatment group. RESULTS: The best estimate of true proportion of CAA and the 95% confidence interval in each group at 30 and 60 days were: (1) ASA group, 30 days, 22.8% (20.6%, 25%); 60 days, 17.1% (13.6%, 20.7%); (2) low-IVIG group, 30 days, 17.3% (14.3%, 20.2%); 60 days, 11.1% (8.7%, 13.6%); (3) high-IVIG group, 30 days, 10.3% (8.3%, 12.3%); 60 days, 4.4% (2.8%, 6%); (4) single-IVIG group, 30 days, 2.3% (0.5%, 4.2%); 60 days, 2.4% (0.5%, 4.2%); (5) high-IVIG-low-ASA group, 30 days, 13% (9%, 17%); 60 days, 4.8% (2.3%, 7.4%); and (6) high-IVIG-high-ASA group, 30 days, 9.1% (6.9%, 11.4%); 60 days, 4% (2%, 6.1%). CONCLUSION: The incidence of CAA both at 30 and 60 days was significantly lower in low-IVIG than in ASA and in high-IVIG than in low-IVIG groups. Also, the incidence was lower in the single-IVIG than in the high-IVIG group, but this was noted at 30 days and not at 60 days. There was no statistically significant difference in the incidence of CAA both at 30 and 60 days between the high-IVIG-low-ASA and high IVIG-high-ASA groups. PMID- 7491222 TI - Psychological response to HIV positivity in hemophilia. AB - OBJECTIVE: To compare the psychological and family adaptation of children and adolescents with hemophilia who were seropositive for human immunodeficiency virus type 1 (HIV-1) with an HIV-negative group of comparable age, demographic characteristics, and disease severity in a multi-site study. DESIGN: Cross sectional, controlled study. SETTING: Thirty-three hemophilia treatment centers throughout the United States. SAMPLE: Ninety-one children and adolescents with hemophilia who were seropositive for HIV and 92 children and adolescents with hemophilia who were seronegative and of comparable age, demographic characteristics, and disease severity. RESULTS: HIV-seropositive children and adolescents reported less positive affect [(lower well being) (P < .05)], and more frequent hemophilia-related school absences were identified among HIV infected patients (P < .005). However, the two groups demonstrated surprisingly comparable levels of psychological, social, hemophilia-related adjustment, general family relations, and hemophilia-related family adaptation, as reported by patients and parents. However, mothers of HIV-seropositive children and adolescents reported higher levels of general psychological distress (P < .008) and higher levels of distress related to hemophilia (P < .0002) than parents of HIV-negative children. CONCLUSIONS: Seropositive children and adolescents with hemophilia demonstrate psychological resilience and levels of psychological adjustment that were comparable to seronegative counterparts. However, mothers of seropositive children were more distressed than mothers of HIV-negative children. Practitioners should ensure that stressed mothers obtain necessary psychological support. PMID- 7491223 TI - Neurological correlates of fetal cocaine exposure: transient hypertonia of infancy and early childhood. AB - OBJECTIVE: To assess whether prenatal cocaine exposure has any long-term effects on neurodevelopment. DESIGN: A prospective cohort study with examiners blind to drug exposure and human immunodeficiency virus (HIV) status. SUBJECTS: Of 144 high-risk infants enrolled in a perinatal HIV neurodevelopmental study, 119 (83%) infants with both neurological and urine toxicology measures were followed up to age 24 months. METHODS: Neurological and developmental assessments were analyzed at 6-month intervals grouped according to the presence of cocaine in urine toxicology: 51 infants were cocaine-positive. Adjusted odds ratios (ORs) and 95% confidence interval (CI) were obtained by logistic regression equations that adjusted for perinatal variables, including measures of fetal growth, gestation, HIV status, and infant toxicology results. SETTING: Harlem Hospital Center from 1988 to 1992. RESULTS: At age 6 months, 21 of 51 (41%) cocaine-positive children exhibited hypertonia of any type (hypertonic tetraparesis, hypertonic diparesis, and hypertonic hemiparesis) compared with 17 of 68 (25%) cocaine-negative infants (OR = 2.1, CI = 1.0-4.6). Cocaine-positive infants were four times more likely to show hypertonic tetraparesis (HTP) than cocaine-negative infants (OR = 4.0; CI = 1.5-10.8). The association remained significant in multivariate analyses. Hypertonia, consistent with cerebral palsy, diminished over time in both groups. In 97% of affected infants hypertonia resolved by 24 months. Arm hypertonia abated first; leg hypertonia remained in some children up to age 18 months. No differences in development scores between cocaine-positive and cocaine-negative were noted at any age interval. However, among cocaine-positive infants those with early HTP showed significantly lower mean developmental scores at 6 and 12 month compared to infants without HTP. CONCLUSION: Cocaine positivity urine toxicology at birth is associated with hypertonia during infancy. Such cocaine induced effects are usually symmetrical, transient, and the majority of exposed children outgrow hypertonia by 24 months of life. Among cocaine-positive infants, HTP may be a marker for later developmental impairments. PMID- 7491224 TI - The effects of HIV on cognitive and motor development in children born to HIV seropositive women with no reported drug use: birth to 24 months. AB - OBJECTIVE: This study documents delays in the mental and motor functioning of infants perinatally infected with human immunodeficiency virus (HIV) while controlling for confounding effects of prenatal drug exposure, ethnicity, socioeconomic status, and maternal separation and death. METHODS: The cognitive and motor development of 126 infants born to nondrug-using, HIV-seropositive Haitian women was assessed at 3-month intervals through 24 months of age using the Bayley Scales of Infant Development. By 18 months of age, 28 of the infants were diagnosed as HIV-infected, and the 98 uninfected infants served as a control group. The infected and uninfected infants did not differ with respect to mean gestational age, birth weight, ethnicity, or rates of maternal separation and death. RESULTS: By 3 months of age, the mean mental and motor scores of the infected infants were significantly lower than those of the uninfected controls. Furthermore, the initial differences between the two groups increased over time, as many of the infected infants became increasingly delayed. Although the infected infants tended to perform more poorly than the uninfected infants, nearly one third of the infected infants exhibited relatively normal cognitive development and half demonstrated relatively normal motor development. CONCLUSIONS: Over the first 24 months of life, the mean rate of development of HIV-infected infants is significantly slower than that of noninfected infants born to seropositive mothers. This occurs even when the effects are not confounded with those of prenatal drug exposure. PMID- 7491225 TI - Failure to identify human immunodeficiency virus-seropositive newborns: epidemiology and enrollment patterns in a predominantly white, nonurban setting. AB - OBJECTIVE: To describe the epidemiology of newborn seroprevalence for human immunodeficiency virus (HIV) in a predominantly white, nonurban population, and to determine the factors associated with enrollment at a regional pediatric acquired immunodeficiency syndrome (AIDS) center serving that population. DESIGN: Retrospective case series of children enrolled at a regional pediatric AIDS center during a 6-year period and comparison with universal blind newborn screening data collected by the state of New York during the same time interval. SETTING: The Pediatric AIDS Center at State University of New York-Health Science Center at Syracuse, which serves as the only source of HIV-related pediatric care for children in a 16-country region of upstate New York totaling 1.8 million population. RESULTS: One hundred thirty-nine HIV-seropositive infants were born in the region during the 6-year study period; complete blind screening data were available for 138. Sixty-five (47%) of these infants were white. Thirty-nine (28%) of 138 had been enrolled at the Pediatric AIDS Center within the first 90 days of life. An additional 22 (16%) were enrolled at older than 90 days of life. The remaining 77 (56%) have never been seen at the center and are presumed to be unidentified. County enrollment rates varied from 0% to 100% and correlated with percent nonwhite births (r = .58; 95% confidence interval, 0.04-0.86). Children in outlying counties were at greater risk for nonenrollment than children from Onondaga County (site of the Pediatric AIDS Center) (adjusted relative risk, 1.38; 95% confidence interval, 1.05-1.85). White infants residing outside of Onondaga County were at the greatest risk of nonenrollment; of 50 seropositive white infants residing outside of Onondaga County, only 7 (14%) were enrolled at the center within the first 90 days of life. CONCLUSIONS: Local demographic factors can skew the racial distribution of HIV-seropositive infants dramatically compared with the national experience. White race and residence in counties away from the medical center each constituted risk factors for nonenrollment at the Pediatric AIDS Center. The epidemiology of HIV in this predominantly white, rural population, coupled with physician practices, probably contributed to low identification and enrollment rates. As the AIDS epidemic spreads into similar populations elsewhere, HIV infection in pregnant women or newborn infants is likely to become progressively harder to detect, unless universal screening is adopted. PMID- 7491226 TI - The prevalence of sexually transmitted diseases in children and adolescents evaluated for sexual abuse in Cincinnati: rationale for limited STD testing in prepubertal girls. AB - OBJECTIVE: To determine the prevalence of Chlamydia trachomatis, Neisseria gonorrhoeae, Trichomonas vaginalis, syphilis, and human immunodeficiency virus (HIV) infection in sexually abused children and to develop selective criteria for sexually transmitted disease (STD) testing in these children in our community. DESIGN: Prospective. SETTING: University-affiliated children's hospital in Ohio. PARTICIPANTS: All children evaluated at our hospital for sexual abuse were eligible. Eight hundred fifty-five children were evaluated over a 1-year period. The study included 704 girls and 151 boys. Children ranged in age from 3 weeks to 18 years old. METHODS AND RESULTS: Standard STD testing (American Academy of Pediatrics recommendations) was defined as serum rapid plasma reagin test, examination for Trichomonas, N gonorrhoeae culture of the throat, rectum, and genitalia and C trachomatis culture of the rectum and genitalia. STD testing in this study was recommended in children with 1) a history of genital discharge or contact with the perpetrator's genitalia, 2) examination findings of genital discharge or trauma, and 3) all adolescents. HIV testing was obtained in children with risk factors for HIV infection, those with contact with a perpetrator with HIV risk factors, or if the family was concerned about HIV acquisition. A total of 423 children were tested for N gonorrhoeae, 415 for C trachomatis, 275 for syphilis, 208 for Trichomonas, and 140 for HIV. Twelve children were determined to have N gonorrhoeae infection, 11 had C trachomatis infection, and four had Trichomonas infection. Overall, the prevalence of STDs in prepubertal girls was 3.2% and 14.6% in pubertal girls. The prevalence of N gonorrhoeae in prepubertal girls with vaginal discharge was 11.1% and 0% in prepubertal girls without discharge (P < .001). C trachomatis infection was diagnosed in 0.8% of prepubertal girls compared with 7.0% of pubertal girls (P < .001). None of the children tested positive for syphilis or HIV and no males had a STD. CONCLUSIONS: In our community, N gonorrhoeae testing in prepubertal girls can be limited to those with a vaginal discharge on examination unless other risk factors are present. The prevalence C trachomatis and Trichomonas in prepubertal girls is low and may be omitted from routine evaluations. All pubertal girls evaluated for sexual abuse should be tested for STDs because of the high prevalence of asymptomatic infection in this patient population. PMID- 7491227 TI - Oxandrolone therapy in constitutionally delayed growth and puberty. Bio Technology General Corporation Cooperative Study Group. AB - BACKGROUND: Male adolescents with constitutional delay of growth and puberty may have significant psychosocial difficulties related to their sexual immaturity and short stature. The purpose of this study was to test the hypothesis that 1 year of oxandrolone therapy would increase growth velocity and thereby improve psychosocial functioning in boys with constitutional delay of growth and pubertal development. METHODS: Forty boys (ages 11 to 14.7 years) with delayed pubertal development and short stature were recruited from the pediatric endocrine clinics of 14 medical centers. The boys were randomized using a block design stratified for age to receive either oxandrolone (0.1 mg/kg daily for 1 year) or an identical-appearing placebo tablet, using a double-masked design. RESULTS: Growth velocity in the oxandrolone-treated boys was significantly greater than in the control boys (9.5 vs 6.8 cm/y). Likewise, the mean height SD score increased 0.41 in the oxandrolone group, whereas it decreased 0.03 in the control group. Those in the oxandrolone group gained 2.4 kg more than those in the placebo group. Mean predicted adult heights did not change in either group. The mean rates of pubertal progression were equivalent in both groups. Self-image (Piers-Harris Self Concept Scale) and social competence (Child Behavior Profile) were normal at baseline in both groups and did not change significantly over the course of the study in either group. No complications of oxandrolone therapy were identified. CONCLUSIONS: This randomized, placebo-controlled trial demonstrates that low-dose oxandrolone can increase both height and weight velocity in boys with delayed puberty safely. Under the conditions of this study, however, the increased growth velocity in the oxandrolone-treated boys was not associated with a greater improvement in psychosocial status compared with the control boys. PMID- 7491228 TI - Life events, cardiovascular reactivity, and risk behavior in adolescent boys. AB - BACKGROUND: Risk behavior contributes to injuries, one of the most important sources of morbidity and mortality in adolescents. Although research has shown that environmental stress makes adolescents more likely to engage in risk behavior and to sustain injuries, the magnitude of these associations has been small. Little is known about the role of individual differences in psychobiologic reactivity to stress in moderating the impact of stressful events. In this study, we examined associations among environmental stressors, cardiovascular reactivity to stress, and the level of risk behavior in adolescent boys. METHODS: Twenty four 14- to 16-year-old boys underwent a laboratory protocol designed to measure responses to psychologically and physically stressful tasks. Changes in heart rate and mean arterial blood pressure were measured serially at standard points in the protocol, and levels of positive and negative life events and recent risk behavior were measured using self-report questionnaires. RESULTS: Neither life events nor cardiovascular reactivity were independently associated with risk behavior. Positive life events and mean arterial blood pressure reactivity significantly interacted, however, in predicting risk behavior (R2 increment = .25). Boys with high reactivity who reported numerous positive life events engaged in markedly less risk behavior than their peers. CONCLUSION: We conclude that adolescents with exaggerated cardiovascular responses to laboratory stressors are associated with less risk behavior in a setting of positive life circumstances. This result suggests that reactivity may exert protective, rather than harmful, influences in some environments. PMID- 7491229 TI - Sudden infant death syndrome in relation to weather and optimetrically measured air pollution in Taiwan. AB - OBJECTIVE: To examine the possible role of weather and air pollution in sudden infant death syndrome (SIDS) and suffocation. METHODS: Poisson regression analysis was carried out to measure the association between daily rates of SIDS per 1000 live births and daily average values of visibility and temperature in Taiwan between 1981 and 1991. The optimetrical measure of air pollution was used to represent pollution over a whole area rather than at a point source. RESULTS: Mortality from SIDS per 1000 live births was 3.3 times greater in the lowest category of visibility on the day of death than in the highest category; this rate ratio was 3.4 for the average visibility during the 9 days before death. Adjustment for population size, season, level of urbanization, incidence of deaths from respiratory tract infections, temperature, air pressure, sunshine, rainfall, relative humidity, and windspeed increased these rate ratios to 3.8 and 5.1, respectively. This suggests that the relationship between air pollution and SIDS is not biased by ecological confounders. For temperature the rate ratios were between 3.3 and 4.0. CONCLUSIONS: Our findings confirm the association of climatic temperature and air pollution with SIDS. PMID- 7491230 TI - Voiding after neonatal circumcision. AB - OBJECTIVE: To determine whether it is necessary to delay discharge of newly circumcised male neonates to observe voiding. SUBJECTS AND METHODS: A prospective study was conducted in 1992 and 1993 of 51 healthy male, newly circumcised neonates between 0 and 10 days of age. The neonates were observed for the time of first voiding after circumcision was performed. RESULTS: All neonates voided after circumcision at a mean age of 5.3 +/- 2.5 hours, and there were no complications noted in the study population. CONCLUSION: Healthy male infants who are circumcised without obvious complications can be expected to void, and it is unnecessary to delay hospital discharge to make this observation. PMID- 7491232 TI - Prognosis with preoperative pulmonary function and lung volume assessment in infants with congenital diaphragmatic hernia. AB - OBJECTIVES: To determine whether fatal pulmonary hypoplasia, as assessed by functional residual capacity (FRC), can be distinguished from other reversible causes of respiratory failure in infants with congenital diaphragmatic hernia (CDH). METHODS: In the present study, 25 term neonates having CDH without other anomalies (mean birth weight +/- SD, 3.25 +/- 0.50 kg) were enrolled prospectively into a protocol evaluating pulmonary function. Lung compliance (CL) and FRC were measured before diaphragmatic repair and compared with the highest oxygenation index (OI) and lowest PaCO2, also obtained preoperatively. Pulmonary function assessment was repeated after diaphragm repair on postoperative days 3 and 7. CL was determined by esophageal manometry and pneumotachography, and FRC was determined by helium dilution. RESULTS: Fifteen infants (60%) survived to hospital discharge. Eighteen (72%) required extracorporeal membrane oxygenation (ECMO) for support, and of these, 8 (44%) survived. PaCO2 was similar preoperatively in infants grouped as survivors without ECMO, survivors with ECMO, and nonsurvivors. In nonsurvivors (all of whom received ECMO), the preoperative OI was significantly higher (51 +/- 21), CL was less (0.11 +/- 0.04 mL/cm of water per kg), and FRC was smaller (4.5 +/- 1.0 mL/kg) than in the survivors who required ECMO (26 +/- 18, 0.18 +/- 0.08 mL/cm of water per kg, and 12 +/- 5 mL/kg, respectively), as well as in the survivors without ECMO, (6 +/- 2, 0.32 +/ 0.16 mL/cm of water per kg, and 15.8 +/- 4 mL/kg, respectively). The group surviving with ECMO had a higher OI than the infants surviving without ECMO. All nonsurviving infants had FRCs of less than 9.0 mL/kg preoperatively. In contrast, only 2 of the 15 survivors had preoperative FRCs less than 9 mL/kg. CONCLUSIONS: The results of this study suggest that preoperative assessment of FRC may predict fatal pulmonary hypoplasia in most infants with CDH. PMID- 7491231 TI - Continuing risk for hepatitis B virus transmission among Southeast Asian infants in Louisiana. AB - OBJECTIVE: Implementation and evaluation of a hepatitis B vaccination program for Southeast Asian infants in Louisiana. METHODS: A baseline seroprevalence survey of hepatitis B virus (HBV) infection in US-born Southeast Asian children was conducted in 1991 before the implementation of a vaccination program. Hepatitis B vaccination and postvaccination serologic testing of survey participants 10 years of age and younger was performed. Eighteen months after the hepatitis B vaccine was integrated into infant immunization schedules in July 1993, a vaccination coverage survey was performed. RESULTS: Baseline serologic testing was conducted on 96% of persons from 225 randomly selected households in a Southeast Asian community in Louisiana. Of 676 US-born children, 28 (4.1%) had chronic HBV infection; 61% of children with chronic HBV infection were born to hepatitis B surface antigen (HBsAg)-negative women. Among children born to HBsAg-negative women, the prevalence of chronic HBV infection increased with age, reaching 7.3% for children 13 to 16 years of age. Children born to HBsAg-negative women and living with carriers were 5.4 times more likely to have evidence of HBV infection than were children who did not live with carriers. Before the survey, only one child had received three doses of hepatitis B vaccine. In July 1993, 43% of Southeast Asian infants 9 to 18 months of age born in Louisiana had received three doses of hepatitis B vaccine. Infants who received immunizations from private providers were more likely to be fully vaccinated than were infants who received services from public sector clinics (prevalence ratio, 2.1; 95% confidence interval, 1.4, 3.1). CONCLUSIONS: HBV transmission occurs throughout childhood in US-born Southeast Asian children, and the prevalence of chronic HBV infection approaches that of the country of origin. Few US-born Southeast Asian children have received hepatitis B vaccine. Because of the high rates of early childhood HBV transmission and the high risk of chronic infection in Asian and Pacific Islander communities, prevention efforts should be enhanced to ensure that all Asian and Pacific Islander infants receive hepatitis B vaccine in the first 12 months of life and that older children are vaccinated. PMID- 7491233 TI - Fasting insulin concentration is related to cardiovascular reactivity to exercise in children. AB - OBJECTIVE: One mechanism through which hyperinsulinemia is linked to hypertension is through its stimulation of sympathetic nervous activity. Thus, insulin concentration may be correlated with indices of sympathetic activity before it is associated with resting blood pressure. We tested this hypothesis by determining the relationship of insulin concentration and sympathetically mediated cardiovascular reactivity to exercise in children. DESIGN: Survey. SETTING: General community. PARTICIPANTS: Volunteer sample of 46 black and white boys and girls, 9 to 11 years of age. INTERVENTIONS: None. Fasting insulin concentration was the main independent variable. MAIN OUTCOME MEASURES: Systolic blood pressure and heart rate during a standard submaximal bout of treadmill exercise, and systolic blood pressure at peak effort. RESULTS: The hypothesis was tested by multiple regression analyses controlled for resting values. Insulin contributed significantly to the regression models for submaximal heart rate (P < .001), submaximal systolic blood pressure (P = .001), and peak systolic blood pressure (P = .006). CONCLUSIONS: Fasting insulin concentration is associated with cardiovascular reactivity to exercise in young children. This supports the hypothesis that the relationship between hyperinsulinemia and hypertension is mediated by sympathetic nervous tone and that the process begins in childhood. Because percent body fat was positively associated with both insulin and cardiovascular reactivity to exercise, prevention of childhood obesity may be a valuable prophylactic measure for these health problems. PMID- 7491234 TI - Using characteristic changes in laboratory values to assist in the diagnosis of hemorrhagic shock and encephalopathy syndrome. AB - OBJECTIVES: (1) To determine whether the laboratory abnormalities in hemorrhagic shock encephalopathy syndrome (HSES) follow a characteristic pattern of evolution; and (2) to describe the clinical characteristics and outcome of this group of children. DESIGN: Retrospective review of hospital records. SETTING: Pediatric intensive care unit in an academic children's hospital. PATIENTS: Seventeen children who received the diagnosis of HSES. MEASUREMENTS: (1) To support the diagnosis of HSES, the following laboratory tests were obtained: creatine phosphokinase, alanine aminotransferase, aspartate aminotransferase, blood urea nitrogen, creatinine, anemia, thrombocytopenia, coagulation abnormalities, and a metabolic acidosis; and (2) patient characteristics at the time of admission and during the illness and patient outcome were recorded. RESULTS: The median time for laboratory abnormalities in HSES to reach the most aberrant levels was 1.2 to 1.4 days. After reaching maximal deviation, the laboratory values gradually returned to normal. The severity of laboratory abnormalities was not predictive of mortality. All patients were comatose, had seizures, and required fluid resuscitation. Respiratory failure occurred in 12 (71%) of 17 patients. Vasopressor infusions were used for 8 (47%). Viral cultures were performed in 15 patients and were positive in 8 (53%). Eleven (64%) patients survived. CONCLUSIONS: The laboratory abnormalities that are characteristic of HSES follow a distinctive pattern of evolution during the course of the illness. Although the sensitivity, specificity, and predictive value of this pattern of laboratory abnormalities are not known, it might provide a useful tool in diagnosing HSES and excluding other illnesses. Viral illness is common in HSES. The severity of illness and mortality in this group of patients is similar to other descriptions of HSES. PMID- 7491237 TI - Can pediatric training manage in managed care? PMID- 7491235 TI - Comparison of immunogenicity and efficacy of rhesus rotavirus reassortant vaccines in breastfed and nonbreastfed children. US Rotavirus Vaccine Efficacy Group. AB - OBJECTIVE: To evaluate whether breastfeeding affected the immunogenicity and/or efficacy of candidate rhesus-human rotavirus reassortant vaccines. METHODS: A total of 989 healthy infants between 4 and 26 weeks of age were enrolled into a 23-center, prospective, randomized, double-masked, controlled study of the safety, immunogenicity, and efficacy of three doses (4 x 10(4) plaque-forming units) of monovalent rhesus-human viral protein 7, or G, serotype 1 reassortant vaccine, (RRV-S1) or tetravalent vaccine (RRV-TV) consisting of rhesus-human reassortant G serotypes 1, 2, and 4, and the parent RRV G serotype 3. Vaccine efficacy was compared in the breastfed and nonbreastfed children as well as seroconversion rates and postvaccination geometric mean titers (GMTs) of neutralizing antibodies to human serotypes 1, 2, 3, and 4, RRV, and immunoglobulin A to RRV. GMTs in the two feeding groups were compared with and without adjustment for age at initiation of vaccination, prevaccination antibody titers, and the age and prevaccination titer interaction. RESULTS: The seroconversion rates to both vaccines by one or more assays were similar for the breastfed and the nonbreastfed groups (RRV-S1, 84% and 85%, respectively; RRV-TV, 94% and 93%, respectively). There were no significant differences in postvaccination GMTs to either vaccine, measured by any serologic assay, in the two feeding groups. The efficacy of the RRV-S1 vaccine was not significantly lower among the breastfed children than the nonbreastfed children (28% and 39%, respectively). RRV-TV, which is the vaccine being further evaluated for licensure, was equally protective in breastfed and nonbreastfed infants (50% and 51%, respectively). Logistic regression analysis, taking into account differences in age at vaccination and day 1 titer, revealed no evidence of differential vaccine efficacy in the two feeding groups for either vaccine. CONCLUSIONS: These results indicate that the RRV-TV vaccine, given as three doses of 4 x 10(4) plaque-forming units, induces similar seroresponses and protection in breastfed and nonbreastfed US children. PMID- 7491236 TI - Serologic responses to Bartonella and Afipia antigens in patients with cat scratch disease. AB - OBJECTIVE: To assess the serologic response to Afipia and Bartonella, previously named Rochalimaea, in patients with cat scratch disease (CSD) and a healthy control group. DESIGN: Prospective, controlled trial. SETTING: Referral clinic and hospitalized patients in a university medical center. PARTICIPANTS: Eighty patients with CSD and 57 healthy control subjects of similar age. MAIN OUTCOME MEASURES: The immune responses to Afipia felis and Bartonella henselae were evaluated by a newly developed enzyme-linked immunosorbent assay (ELISA) in patients with CSD and healthy control subjects. Responses to B henselae were also measured by indirect fluorescent antibody (IFA) tests. Antibody levels to Bartonella quintana were measured by ELISA and IFA in a limited number of patients and control subjects. RESULTS: Of the 80 patients with clinical CSD, 56 had positive results of CSD skin tests. ELISA antibody levels to A felis did not differ between patients and control subjects, but immunoglobulin M (IgM) and IgG ELISA antibodies to B henselae and B quintana were significantly higher in patients than in control subjects. IFA responses to B henselae and B quintana were also significantly higher in patients than in control subjects. CONCLUSION: Patients with CSD had significant serologic responses to B henselae and B quintana but not to A felis, suggesting that the causative agent of CSD is antigenically related to the Bartonella genus and not to Afipia. The Bartonella IgM ELISA and IFA assay were both sensitive and specific and may be used to establish the diagnosis of CSD. PMID- 7491238 TI - Managed care, re-engineering and downsizing: will medical education survive change? PMID- 7491239 TI - Inhaled nitric oxide for persistent pulmonary hypertension of the newborn: implications and strategy for future "high-tech" neonatal clinical trials. PMID- 7491240 TI - The preparticipation sports physical: are we dropping the ball? PMID- 7491241 TI - Inhaled nitric oxide for persistent pulmonary hypertension of the newborn: the physiology matters! PMID- 7491243 TI - Alveolar capillary dysplasia--a cause of persistent pulmonary hypertension unresponsive to a second course of extracorporeal membrane oxygenation. PMID- 7491244 TI - A simple maneuver to provoke benign neonatal sleep myoclonus. PMID- 7491242 TI - Assessment of glomerular filtration rate in preterm infants by serum creatinine: comparison with inulin clearance. PMID- 7491245 TI - The progression of human papillomavirus lesions in sexual assault victims. PMID- 7491247 TI - Headband alopecia. PMID- 7491246 TI - Health supervision for children with Turner syndrome. American Academy of Pediatrics. Committee on Genetics. PMID- 7491248 TI - Carnitine deficiency. PMID- 7491249 TI - Carnitine deficiency. PMID- 7491250 TI - Clonidine and sudden death. PMID- 7491251 TI - Correct dosing of vancomycin in infants and children. PMID- 7491252 TI - Localization and regulation by steroids of the alpha, beta and gamma subunits of the amiloride-sensitive Na+ channel in colon, lung and kidney. AB - Polyclonal antibodies have been raised against the alpha, beta and gamma subunits of the amiloride-sensitive Na+ channel. The three subunits were detected by immunohistochemistry at the apical membrane of epithelial cells from the distal colon, the lung and the distal segments of the kidney tubules. No significant labelling was detected in lung alveoli, suggesting that it is not a major site of expression of the Na+ channel. Effects of a low Na+ diet or of dexamethasone treatment were measured at the mRNA level and at the protein level by immunohistochemistry. In the colon, steroids controlled Na+ channel activity via the stimulation of the transcription of beta and gamma subunits. The alpha mRNA was constitutively expressed. However, while neither alpha, beta nor gamma proteins were detected in the colon of control animals, they were all detected in the colon of steroid-treated animals. In the lung, Na+ channel expression was regulated by glucocorticoids the circulating level of which was sufficiently high to induce a maximal expression of the three subunits, even in control animals. Adrenalectomy drastically reduced expression of the three subunits. A surprising finding was the apparent absence of steroid effects on alpha, beta and gamma subunit expression in the kidney. Neither the expression of the mRNAs nor the expression of the proteins were significantly altered by aldosterone or by dexamethasone. These results could be due to mixed gluco- and mineralocorticoid regulations in different segments of the kidney tubule, but their interpretation also requires regulations that are apparently not found in the lung or colon. PMID- 7491253 TI - Different modulation of Ca-activated K channels by the intracellular redox potential in pulmonary and ear arterial smooth muscle cells of the rabbit. AB - We investigated the electrical responses of Ca-activated K (KCa) currents induced by hypoxia and reduction or oxidation of the channel protein in pulmonary (PASMC) and ear (EASMC) arterial smooth muscle cells using the patch-clamp technique. In cell-attached patches, in the presence of a high K solution (containing 0.316 microM Ca2+), the activity of KCa channels from PASMC was decreased (by 49 +/- 7% compared to control, pipette potential = -70 mV) by changing to a hypoxic solution (1 mM Na2S2O4, aeration with 100% N2 gas). EASMC channels did not respond to hypoxia. In order to investigate the possible mechanisms involved, using inside-out patches bathed symmetrically in 150 mM KCl, we applied redox couples to the intracellular side. Reducing agents, such as dithiothreitol (DDT, 5 mM), reduced glutathione, (GSH, 5 mM), and nicotinamide adenine dinucleotide reduced (NADH, 2 mM) decreased PASMC, but not EASMC, KCa channel activity. However, oxidizing agents such as 5,5'-dithio-bis(2-nitrobenzoic acid) (DTNB, 1 mM), oxidized glutathione (GSSG, 5 mM) and NAD (2 mM) increased KCa channel activity in both PASMC and EASMC. The increased activity due to oxidizing agents was restored by applying reducing agents. From these results, we could suggest that the basal redox state of the EASMC KCa channel is more reduced than that of the PASMC channel, since the response of KCa channels of the EASMC to intracellular reducing agents differs from that of the PASMC. This difference may be related to the different responses of PASMC and EASMC KCa channels to hypoxia. PMID- 7491254 TI - Calcium and barium permeation through calcium release-activated calcium (CRAC) channels. AB - A Ca2+ current activated by store depletion has been described recently in several cell types and has been termed ICRAC (for Ca2+ release-activated Ca2+ current). In this paper, the Ca2+ and Ba2+ permeability of CRAC channels is investigated in mast cells, rat basophilic leukaemia cells (RBL) and human T lymphocytes (Jurkat). The selectivity of CRAC channels for Ca2+ over monovalent cations is identical in all three cell types and is at least as high as that of voltage-operated Ca2+ (VOC) channels in the various tissues tested. The amplitude of Ba2+ currents relative to Ca2+ currents (IBa/ICa) through CRAC channels was found to be strongly dependent on the membrane potential and was much smaller in Jurkat cells compared to mast and RBL cells. An anomalous mole-fraction behavior was observed at very negative membrane potentials in all three cell types when using different mixtures of external Ca2+ and Ba2+. In contrast to VOC channels, the anomalous mole-fraction effect was not observed at potentials positive to -20 mV. PMID- 7491256 TI - Ca(2+)- and GTP[gamma S]-induced translocation of the glucose transporter, GLUT 4, to the plasma membrane of permeabilized cardiomyocytes determined using a novel immunoprecipitation method. AB - In cardiomyocytes glucose transport is activated not only by insulin but also by contractile activity that causes translocation of the glucose transporter, GLUT 4, from intracellular vesicles to the plasma membrane. The latter effect may possibly be mediated by intracellular Ca2+, as suggested by previous studies. To investigate the role of Ca2+, we permeabilized neonatal rat myocytes with alpha toxin and incubated them for 1 h either at a pCa (i.e.--log10 [Ca2+]) of 8 (control) or at a pCa of 5 in the presence of adenosine 5'-triphosphate (ATP). Translocation of GLUT-4 was then monitored by a novel immunoprecipitation method using a peptide antibody directed against an exofacial (extracellular) loop of GLUT-4 (residues 58-80). Incorporation of GLUT-4 into the plasmalemma was stimulated 1.8-fold by 10 microM Ca2+ and 1.7-fold by insulin (as in the case of intact cells). The insulin effect was Ca2+ independent, i.e. it was identical in the absence and presence of Ca2+ (10 microM). Guanosine 5'-O-(3-thio triphosphate) (GTP[gamma S]), which was inactive in intact cells, also caused translocation of GLUT-4 in permeabilized cardiomyocytes. Thus, incorporation of GLUT-4 into the plasma membrane was enhanced 2.5-fold by 200 microM GTP[gamma S] in the virtual absence of Ca2+ (pCa 8) and even 3.5-fold at 10 microM free Ca2+. We conclude that an increase in intracellular Ca2+ concentration increases GLUT-4 translocation of (permeabilized) cardiomyocytes to a similar extent as do insulin and GTP[gamma S] in the absence of Ca2+, but that the effects of Ca2+ and GTP[gamma S] may be additive. PMID- 7491255 TI - Regulation of the L-type calcium channel alpha-1 subunit by chronic depolarization in the neuron-like PC12 and aortic smooth muscle A7r5 cell lines. AB - The regulation of L-type voltage-dependent Ca2+ channels by chronic depolarization was studied in the aortic smooth muscle A7r5 and neuron-type PC12 cell lines, by probing the expression and the functional state of their constitutive alpha-1 subunits. PC12 cells showed, after prolonged exposure to a high-K+ depolarizing solution, a 25% reduction of the functional Ca2+ channel density which was accompanied by a decrease of the alpha-1 subunit mRNA expression. In A7r5 cells submitted to a similar protocol of depolarization, 45Ca2+ uptake measurements revealed a fall in the functional activity of L-type Ca2+ channels which was not related to a modulation of their mRNA expression, but arose from a long-term voltage-dependent channel inactivation. Accordingly, the lag time and the mechanisms of recovery were different in the two cell types. In PC12 cells, when restoring physiological culture conditions, de novo synthesis of alpha-1 subunits allowed the recovery of the original density of L-type Ca2+ channels at the membrane surface. As for the A7r5 cells, we showed that after chronic depolarization, the complete restoration of the resting membrane potential and the related Ca2+ channel activity required a 2-day incubation in physiological medium and could probably be related to a normalization of the increased intracellular Ca2+ concentration. In contrast, it is noteworthy that, in PC12 cells, the only transient increase of intracellular Ca2+ content in the first hours of depolarization could account for the long-term down-regulation of L-type Ca2+ channels. PMID- 7491257 TI - On the regulation of the expressed L-type calcium channel by cAMP-dependent phosphorylation. AB - The Ca2+ channel subunits alpha 1C-a and alpha 1C-b were stably expressed in Chinese hamster ovary (CHO) and human embryonic kidney (HEK) 293 cells. The peak Ba2+ current (IBa) of these cells was not affected significantly by internal dialysis with 0.1 mM cAMP-dependent protein kinase inhibitor peptide (mPKI), 25 microM cAMP-dependent protein kinase catalytic subunit (PKA), or a combination of 25 microM PKA and 1 microM okadaic acid. The activity of the alpha 1C-b channel subunit expressed stably in HEK 293 cells was depressed by 1 microM H 89 and was not increased by superfusion with 5 microM forskolin plus 20 microM isobutyl methylxanthine (IBMX). The alpha 1C-a.beta 2.alpha 2/delta complex was transiently expressed in HEK 293 cells; it was inhibited by internal dialysis of the cells with 1 microM H 89, but was not affected by internal dialysis with mPKI, PKA or microcystin. Internal dialysis of cells expressing the alpha 1C a.beta 2.alpha 2/delta channel with 10 microM PKA did not induce facilitation after a 150-ms prepulse to +50 mV. The Ca2+ current (ICa) of cardiac myocytes increased threefold during internal dialysis with 5 microM PKA or 25 microM microcystin and during external superfusion with 0.1 microM isoproterenol or 5 microM forskolin plus 50 microM IBMX. These results indicate that the L-type Ca2+ channel expressed is not modulated by cAMP-dependent phosphorylation to the same extent as in native cardiac myocytes. PMID- 7491258 TI - Barium block of the muscarinic potassium current in guinea-pig atrial cells. AB - Block of the muscarinic K+ current (iK,ACh) by Ba2+ has been studied in guinea pig atrial cells using the whole-cell patch-clamp technique. The dose-response curve for the block of iK,ACh can be fitted assuming that a muscarinic K+ channel is blocked when a single Ba2+ ion binds to it (apparent dissociation constant, Kd = 125 microM at 0 mV). Block was voltage and time dependent. The voltage dependence can be explained by Ba2+ binding to a site within the pore of the channel, 36% across the width of the membrane electric field (from the outside). Raising the bathing K+ concentration reduced Ba2+ block of iK,ACh, which suggests that Ba2+ and K+ compete for a common binding site. When Ba2+ was added during an exposure to ACh (muscarinic K+ channel open), block of iK,ACh developed rapidly, but when Ba2+ was added prior to an exposure to ACh (muscarinic K+ channel closed), little block of iK,ACh was evident when ACh was first applied. This suggests that when the muscarinic K+ channel is closed in the absence of ACh, Ba2+ does not have access to the binding site within the pore of the channel. In conclusion, Ba2+ block of iK,ACh is concentration, voltage, time, K+ and state dependent. PMID- 7491259 TI - Modulation of the ryanodine receptor sarcoplasmic reticular Ca2+ channel in skinned fibers of fast- and slow-twitch skeletal muscles from rabbits. AB - This study was performed to compare skinned fibers from rabbit adductor magnus (AM) and soleus (SL) muscles with regard to the influence of caffeine, Ca2+ and Mg2+ on the depressive effects of ryanodine (RYA) on the caffeine-induced tension transients. Single skinned fibers were immersed in solutions to load Ca2+ into, and release Ca2+ from the SR (a load-release cycle). Three cycles were sequentially performed in each skinned fiber: (1) a control (no RYA), (2) a conditioning period in which activation was carried out in the presence of ryanodine plus various concentrations of the modulators, i.e. caffeine, Ca2+ or Mg2+, and (3) a test (no RYA) which monitored the release activity retained after the conditioning cycle. The depressive effect of RYA was found to be a function of [ryanodine], [caffeine], or [Ca2+], and an inverse function of [Mg2+], where [] denotes concentration. The half-maximal effects of RYA in AM (5 microM RYA) and SL (10 microM RYA), respectively, occurred at a pCa50 of 5.32 versus 5.43 without caffeine, or pCa50 of 7.24 versus 6.88 and pMg50 of 3.29 versus 3.61 with 25 mM caffeine, at a [caffeine] of 4.96 versus 7.29 mM, and at a [ryanodine] of 31.0 versus 101.6 microM. Thus, the RYA depression in skinned muscle fibers is modulated by caffeine, Ca2+, and Mg2+ in both muscle types, and AM is at least two- to fourfold more sensitive than SL. PMID- 7491261 TI - Kinetics of receptor-mediated endocytosis of albumin in cells derived from the proximal tubule of the kidney (opossum kidney cells): influence of Ca2+ and cAMP. AB - In this study we investigated the effects of Ca2+ and cyclic adenosine monophosphate (cAMP) on the kinetic of receptor-mediated (RME) and fluid-phase (FPE) endocytosis in opossum kidney (OK) cells, derived from the proximal tubule of the kidney. We used fluorescein isothiocyanate (FITC)-labelled albumin and FITC-labelled dextran as endocytotic substrates for RME and FPE, respectively. Removal of extracellular Ca2+ led to a dramatic decrease of the apparent affinity of RME, but did not influence the maximum endocytotic uptake rate (Jmax). Reduction of extracellular Ca2+ to 1 mumol/1 had no effect. Apparent affinity of specific binding of albumin to the plasma membrane was increased to 200% of control in the absence of extracellular Ca2+, whereas maximum binding capacity was slightly decreased. FPE was not affected by removal of extracellular Ca2+. Additional removal of cytoplasmic Ca2+, using ionomycin, had no further effect on RME and did not affect FPE. Increases of cytoplasmic (using ionomycin at extracellular Ca2+ concentrations of 1 mumol/l or 1.2 mmol/l) or extracellular Ca2+ did not alter the kinetics of RME or FPE. Dibutyryl-cAMP reduced Jmax but left the apparent affinity of RME unchanged. FPE and albumin binding to the plasma membrane were not changed in the presence of cAMP. Removal of extracellular Ca2+ and addition of cAMP led to an alkalinization of endocytotic vesicles. Yet the alkalinization induced by removal of Ca2+ was significantly greater as compared to the alkalinization in the presence of cAMP. Endosomal alkalinization with bafilomycin A1 had no further effect in the absence of Ca2+, but reduced RME in the presence of cAMP.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7491260 TI - Direct measurements of Ca(2+)-activated K+ currents in inner hair cells of the guinea-pig cochlea using photolabile Ca2+ chelators. AB - Intracellular photorelease of Ca2+ from caged Ca2+ (DM-nitrophen or nitr5) and the patch-clamp technique in the whole-cell configuration were used to investigate Ca(2+)-activated currents in inner hair cells (IHCs) of the mammalian cochlea. Photoliberation of intracellular Ca2+ activated outward currents with a mean amplitude of 260 +/- 110 pA when IHCs were voltage-clamped, near the resting membrane potential, at -50 mV. The photoactivated currents were reversibly blocked by extracellular application of tetraethylammonium (TEA, 10 mM), neomycin (1 mM) and charybdotoxin (1 microM), but not by apamin. The voltage dependence of membrane currents activated by photolysis of DM-nitrophen demonstrated a reversal potential near the K+ equilibrium potential (Ek) and saturation near 0 mV. The presence of Ca(2+)-activated currents was further confirmed by the effects of extracellular adenosine 5'-triphosphate (ATP, 10 microM) and the Ca2+ ionophore ionomycin (10 microM). Both agents raised intracellular Ca2+ and simultaneously activated outward currents when IHCs were voltage-clamped near the resting membrane potential. In experiments where currents were activated by depolarizing voltage steps, nifedipine (50 microM) and Cd2+ (1 mM) reduced significantly (20 50%) the whole-cell outward currents, suggesting the presence of L-type Ca2+ currents activating K+ currents. These results are the first direct evidence for Ca(2+)-activated K+ currents in mammalian IHCs, these currents being potentially important for cell repolarization during sound-induced depolarization and synaptic transmission. PMID- 7491262 TI - Expression of functional GABAA receptors in neuroendocrine gastropancreatic cells. AB - Gastropancreatic neuroendocrine cells synthesize large amounts of gamma aminobutyric acid (GABA). This amino acid neurotransmitter appears to be stored in and released from, vesicles similar to small synaptic vesicles. So far, the function of GABA in gastropancreatic, neuroendocrine cells has not been clarified. Previous work suggested that only pancreatic, glucagon-producing alpha 2 cells contain functional GABAA receptors. Using subunit-specific antibodies in sections of human antral mucosa, a human gastrinoma and rat pancreas, we show that expression of GABAA receptors is abundant in gastropancreatic, neuroendocrine cells. Using the patch-clamp technique in the whole-cell mode we demonstrate that both the rat insulinoma cell line RIN 38 and the amphicrine cell line AR42J express functional GABAA receptors, which are characterized by a relatively low benzodiazepine and Zn2+ sensitivity and by an insensitivity to the inverse benzodiazepine agonist 6,7-alpha-methoxy-4-ethyl-beta-carboline-3 carboxylate (DMCM). In contrast to neurons, activation of GABAA receptors leads to a membrane depolarization. This depolarization presumably activates voltage gated Ca2+ channels, resulting in an increase in cytosolic Ca2+ concentration, [Ca2+]i, as shown with the fluorimetric dye fura-2. The combination of GABA release, GABAA receptor activation and the [Ca2+]i increase could constitute an autocrine mechanism, modulating the release of hormones such as gastrin, insulin and somatostatin. PMID- 7491263 TI - Effects of beta-guanidinopropionic acid-feeding on the patterns of myosin isoforms in rat fast-twitch muscle. AB - Administration of beta-guanidinopropionic acid (beta-GPA) to rats as 1% of their diet for 6 weeks led to an accumulation of beta-GPA and beta-GPA-phosphate and to a depletion of creatine and phosphocreatine in the fast-twitch plantaris muscle. Adenosine triphosphate concentration was also decreased. Electrophoretic analyses were performed to investigate the effects of beta-GPA on the patterns of fast (FM) and slow (SM) isomyosins, myosin heavy chain (HC) isoforms and myosin light chain (LC) isoforms. The relative concentrations of fast isomyosins FM1 and FM2 decreased, whereas slow isomyosin SM increased. The increase in slow isomyosin corresponded to an increase in the relative concentration of the slow myosin HCI. The changes of the myosin light chain pattern consisted of increases in the relative concentrations of the two slow isoforms, LC1sb and LC2s, and decreases in the fast isoforms LC2f and LC3f. These results demonstrate that beta-GPA administration, leading to a depletion in energy-rich phosphates and a reduced phosphorylation potential, has an impact on myosin isoform expression in rat fast twitch skeletal muscle. PMID- 7491264 TI - H+/di-tripeptide transporter (PepT1) expression in the rabbit intestine. AB - In order to examine the intestinal expression of the recently cloned H+/di tripeptide transporter (PepT1), oligonucleotide probes were synthesized and their specificity confirmed by Northern blot analysis of rabbit jejunal RNA. In situ hybridization studies, using these probes, show that PepT1 is expressed all along the small intestine and at a very much reduced level in the colon. In contrast, PepT1 mRNA was not detected in the stomach, sacculus rotundus or caecum. Microscopic examination of tissue sections showed PepT1 expression to be restricted to intestinal epithelium with no detectable expression in the lamina propria, muscularis mucosae, muscularis or serosa. The accumulation of PepT1 mRNA along the crypt-villus axis was also investigated. In all regions of the small intestine (in duodenum, jejunum and ileum), PepT1 mRNA was undetectable in deeper epithelial cells of the crypts. Expression was first detectable at or near the crypt-villus junction, the amount of PepT1 mRNA increasing rapidly in the lower villus to a maximum approximately 100-200 microns from this point. Along the length of the small intestine PepT1 mRNA was most abundant in duodenal and jejunal enterocytes, with lower levels in the ileal epithelium. PepT1 expression is greatly depressed in the follicle-associated epithelium of the Peyer's patch relative to both interfollicular and adjacent "normal" villi. These data are discussed in the context of the known physiological role of PepT1 in the gastrointestinal tract. PMID- 7491265 TI - ADP exerts a protective effect against rundown of the Ca2+ current in bovine chromaffin cells. AB - In isolated chromaffin cells, the high-voltage-activated Ca2+ current, recorded using 5 mM Ca2+ as the divalent charge carrier, exhibits rundown within 10 min, which is delayed for 1 h at least by the addition of 1 mM adenosine 5' triphosphate (ATP) to the pipette medium. The mechanism of this stabilizing action of ATP has been examined. ATP action is dose dependent; the rundown process, which was delayed at concentrations below 0.4 mM, was totally abolished at higher concentrations. The requirement for ATP was shown to be quite strict: 2 mM inosine 5'-triphosphate (ITP) could not replace ATP, whereas guanosine 5' triphosphate (GTP) could, but at higher concentrations. This effect of ATP was shown to require the presence of MgCl2 and the liberation of a phosphate group since the ATP analogue 5'-adenylyl-imidodiphosphate (AMP-PNP) could not act as a substitute for ATP, suggesting an action through either adenosine 5'-diphosphate (ADP) or a phosphorylation step. ADP, in the presence of Mg2+ only, could replace ATP in the same concentration range. This effect was shown to be specific to ADP; it was maintained after blocking the pathways which convert ADP into ATP, and could not be mimicked by guanosine 5'-diphosphate (GDP). Similarly, ATP and ADP effects were abolished at an increased internal Ca2+ concentration (pCa 6 instead of pCa 7.7, where pCa = -log10[Ca2+]). Nevertheless, the presence of 1 mM Mg-ADP in the bathing solution did not prevent the rundown of the Ca2+ channels when going to the inside-out patch recording configuration.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7491267 TI - 3-Isobutyl-1-methylxanthine (IBMX) affects potassium permeability in rat sensory neurones via pathways that are sensitive and insensitive to [Ca2+]in. AB - The effects of externally applied 3-isobutyl-1-methylxanthine (IBMX), in millimolar concentrations, on the membrane currents in dorsal root ganglia (DRG) neurones isolated from newborn rats were investigated using the amphotericin based "perforated" patch-clamp technique. In some experiments, simultaneous measurements of intracellular Ca2+ concentration ([Ca2+]in) were performed using fura-2 microfluorimetry. Applications of IBMX induced elevation of [Ca2+]in resulting from Ca2+ release from caffeine-ryanodine-sensitive internal stores. In addition to Ca2+ release, IBMX produced a biphasic membrane current response comprised of an inward current transiently interrupted by outward current. The onset of the inward current slightly preceded the onset of the [Ca2+]in transient, while the interrupting outward current developed synchronously with the [Ca2+]in rise. The development of IBMX-induced outward current ultimately needed the [Ca2+]in elevation. After the depletion of Ca2+ stores by IBMX or caffeine exposure, the subsequent IBMX challenge failed to produce both the [Ca2+]in transient and outward membrane current, although the inward current remained unchanged. Both components of the IBMX-induced membrane current response had a reversal potential close to the K+ equilibrium potential and the IBMX induced membrane current response disappeared while dialysing the cell interior with K(+)-free, Cs(+)-containing solutions suggesting their association with K+ channel activity. External administration of 10 mM tetraethylammonium chloride (TEA-Cl) evoked an inward current similar to that observed in response to IBMX; in the presence of TEA-Cl, IBMX application was almost unable to induce additional inward current. IBMX (5 mM) effectively (approximately 50%) inhibited K+ currents evoked by step depolarizations of membrane potential.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7491266 TI - ATP and G proteins affect the runup of the Ca2+ current in bovine chromaffin cells. AB - The Ca2+ current recorded by the whole-cell technique in chromaffin cells shows, before the often described rundown, a transient facilitation or runup. Initial current amplitude was 570 +/- 165 pA and then it increased by 49 +/- 23% (n = 19, SD) over 2 +/- 1 min in the absence of adenosine 5'-triphosphate (ATP). In the presence of ATP, this process occurred with the same magnitude but it was slowed in a dose-dependent manner, lasting 17 +/- 2 min with 2 mM ATP (n = 8). Since adenosine 5'-diphosphate (ADP) does not reproduce this ATP effect, a complex series of phosphorylations is likely to intervene and we show that, at least, a cAMP-dependent i.e., cyclic adenosine monophosphate) phosphorylation occurs. Pertussis toxin (PTX) pretreatment yielded an already maximal Ca2+ current (around 1000 pA) at the time of the patch rupture, which only slightly increased thereafter (10%, n = 11). Also, guanosine 5'-diphosphate (GDP) and guanosine 5'-O (2-thiodiphosphate) (GDP[ beta s]), induced a fast runup, which was absent in the presence of GTP. Furthermore, we show that facilitation does not occur in the presence of dihydrophyridine (DHP) antagonists. Globally, our data suggest that an ATP-dependent phosphorylation stabilizes the inhibitory control exerted by a PTX-sensitive G protein and, as a result, slows down the facilitation of L-type Ca2+ channels. The recruitment of L-type channels can also be facilitated by the application of a DHP agonist or a depolarizing prepulse protocol.l We show that these processes are only effective over a period which parallels the runup and are not additive to it.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7491268 TI - Mechanism of the ATP-induced rise in cytosolic Ca2+ in freshly isolated smooth muscle cells from human saphenous vein. AB - Effects of exogenous adenosine 5'-triphosphate (ATP) were studied by measurements of intracellular Ca2+ concentration ([Ca2+]i) and membrane currents in myocytes freshly isolated from the human saphenous vein. At a holding potential of -60 mV, ATP (10 microM) elicited a transient inward current and increased [Ca2+]i. These effects of ATP were inhibited by alpha,beta-methylene adenosine 5'-triphosphate (AMP-CPP, 10 microM). The ATP-gated current corresponded to a non-selective cation conductance allowing Ca2+ entry. The ATP-induced [Ca2+]i rise was abolished in Ca(2+)-free solution and was reduced to 30.1 +/- 5.5% (n = 14) of the control response when ATP was applied immediately after caffeine, and to 23.7 +/- 3.8% (n = 11) in the presence of thapsigargin. The Ca(2+)-induced Ca2+ release blocker tetracaine inhibited the rise in [Ca2+]i induced by both caffeine and ATP, with apparent inhibitory constants of 70 microM and 100 microM, respectively. Of the ATP-induced increase in [Ca2+]i 29.3 +/- 3.9% (n = 8) was tetracaine resistant. It is concluded that the effects of ATP in human saphenous vein myocytes are only mediated by activation of P2x receptor channels. The ATP induced [Ca2+]i rise is due to both Ca2+ entry and Ca2+ release activated by Ca2+ ions that enter the cell through P2x receptor channels. PMID- 7491271 TI - Regulation of opossum kidney (OK) cell Na/Pi cotransport by Pi deprivation involves mRNA stability. AB - Renal proximal tubular Na-dependent phosphate transport (Na/Pi cotransport) has been studied extensively in the opossum kidney (OK) cell line. Recently, we cloned a complementary deoxyribonucleic acid (cDNA) (NaPi-4) from OK cells encoding an apical NaPi cotransport system. OK cells exposed to a low-Pi medium, as compared to high-Pi media, responded with an increase in Na/Pi cotransport, which was followed by an increase in NaPi-4 messenger ribonucleic acid (mRNA) abundance; maximal stimulation of Na/Pi cotransport was reached in 2 h, with no further increase for up to 16 h. NAPi-4 mRNA abundance was unaltered for 2 h, then increased to a maximum after 6-16 h in cells treated with low Pi medium. NaPi-4 mRNA decay rate was lowered by low-Pi media when compared to high-Pi media, with no increase in the NaPi-4 mRNA transcription rate. These data suggest that the upregulation of Na/Pi cotransport in OK cells by low-Pi media involves two regulatory mechanisms: an immediate (early) increase (after 2 h) in the expression of Na/Pi cotransport, independent of mRNA synthesis or stability, and a delayed (late) effect (after 4-6 h), resulting in an increase in NaPi-4 mRNA abundance, due to an increased stability. PMID- 7491270 TI - Evidence for a glycerol pathway through aquaporin 1 (CHIP28) channels. AB - Permeabilities to glycerol and small non-electrolytes of three Aquaporin 1 CHIP (AQP1) water channels were measured in AQP1 cRNA-injected Xenopus laevis oocytes and in human AQP1 channels reconstituted in proteoliposomes. By an "osmotic" swelling assay, significant increases of ethylene glycol, glycerol and 1,3 propanediol apparent permeability coefficients (P'solutes) were found in oocytes expressing human, rat and frog AQP1. p-Chloromercuribenzene sulphonate (pCMBS) and CuSO4 inhibited, by 95% and 58% respectively, apparent glycerol permeability (P'gly) in oocytes expressing human AQP1. pCMBS inhibition was reversed by beta mercaptoethanol and CuSO4 inhibition was partly reversed by the Cu(2+)-binding peptide Gly-Gly-His. Tritiated glycerol uptakes confirmed the augmented P'gly value of AQP1 cRNA-injected oocytes. In contrast, no increases of urea, meso erythritol, D-or L-threitol, xylitol and mannitol uptakes were detected. Stopped flow light scattering experiments performed with human AQP1 proteoliposomes also revealed a much greater increase of P'gly than did those with protein-free liposomes; the initial rate of proteoliposomes also swelling was inhibited by 96.2% with HgCl2 and by 72.5% with CuSO4. In AQP1 cRNA-injected oocytes and in proteoliposomes, the value of the glycerol reflection coefficient was 0.74-0.80, indicating that water and glycerol share the same pathway. All these results provide strong evidence that water and certain small solutes permeate the AQP1 channels expressed at the surface of X. laevis oocytes or reconstituted in proteoliposomes. The urea exclusion suggests that the selectivity of the AQP1 channels not only depends on the size of the solutes but probably also on their flexibility and their ability to form H-bonds. PMID- 7491269 TI - Interaction of the antiepileptic drug lamotrigine with recombinant rat brain type IIA Na+ channels and with native Na+ channels in rat hippocampal neurones. AB - Actions of the new antiepileptic drug lamotrigine (LTG, Lamictal) were characterised using recombinant rat brain type IIA Na+ channels expressed in Chinese hamster ovary (CHO) cells and native Na+ channels in rat hippocampal pyramidal neurones, using whole-cell recording and intracellular recording techniques. In CHO cells, LTG caused a tonic inhibition of Na+ currents in a concentration-dependent and voltage-dependent manner. The half-maximal inhibitory concentration (IC50) of approximately 500 microM was obtained at a holding potential (Vh) of -90 mV compared with an IC50 of 100 microM at a Vh of -60 mV. LTG (50 microM) caused a 10-mV negative shift in the slow, steady-state inactivation curve and delayed considerably the recovery from inactivation, but had no significant effects on the voltage dependence of activation or fast inactivation, suggesting that LTG acts mainly on the slow inactivated state. The affinity for the inactivated channels was estimated at 12 microM. The tonic inhibition was augmented by a use-dependent action in which a further inhibition by the drug developed during rapid repetitive stimulation using a train of 20-ms duration pulses (11 Hz). These results were consistent with the drug action being on firing properties of pyramidal neurones. Only in those epilepti-form bursts which caused cumulative inactivation of Na+ spikes did LTG produce a potent inhibition. Our data suggest that the inactivated channel is a primary target for LTG action at therapeutic concentrations. PMID- 7491272 TI - Antisense oligonucleotides suppress cell-volume-induced activation of chloride channels. AB - Cell volume regulation is an essential feature of most cells. After swelling in hypotonic media, the simultaneous activation of potassium and chloride channels is believed to be the initial, time-determining step in cell volume regulation. The activation of both pathways is functionally linked and enables the cells to lose ions and water, subsequently leading to cell shrinkage and readjustment of the initial volume. NIH 3T3 fibroblasts efficiently regulate their volume after swelling and bear chloride channels that are activated by decreasing extracellular osmolarity. The chloride current elicited in these cells after swelling is reminiscent of the current found in oocytes expressing an outwardly rectifying chloride current termed ICln. Introduction of antisense oligodeoxynucleotides complementary to the first 30 nucleotides of the coding region of the ICln channel into NIH 3T3 fibroblasts suppresses the activation of the swelling-induced chloride current. The experiments directly demonstrate an unambiguous link between a volume-activated chloride current and a cloned protein involved in chloride transport. PMID- 7491273 TI - Effects of cold exposure and hibernation on renal Na,K-ATPase of the jerboa Jaculus orientalis. AB - Changes in activity and abundance of renal Na,K-ATPase were evaluated during cold exposure and hibernation of the jerboa Jaculus orientalis by measuring the hydrolytic activity, the number of units and the transport activity of Na,K ATPase in isolated nephron segments. As compared to controls, jerboas exposed to cold (6 degrees C) for 4-5 weeks displayed mild diuresis, decreased urinary osmolality and increased kaliuresis. In cold-exposed jerboas, Na,K-ATPase hydrolytic activity was reduced in the medullary thick ascending limb and enhanced in the cortical and outer medullary collecting duct, whereas it was not altered in other nephron segments. The number of Na,K-ATPase units and the activity of Na,K-pump, determined by [3H]-ouabain binding and by ouabain sensitive rubidium uptake respectively, changed in parallel with the hydrolytic activity in the medullary thick ascending limb and cortical collecting duct. The maximal rate of activity (Vmax) of Na,K-ATPase was not modified further during hibernation. Thus, cold exposure, but not the onset of hibernation, induces segment-specific changes in the abundance and activity of Na,K-ATPase units which are likely to be related to the entry into hibernation, but not to the maintenance of some renal functions during deep hibernation. PMID- 7491274 TI - Luminal transport system for H+/organic cations in the rat proximal tubule. Kinetics, dependence on pH; specificity as compared with the contraluminal organic cation-transport system. AB - The efflux of radiolabelled organic cations from the tubular lumen into proximal tubular cells was investigated by using the stop-flow microperfusion method. The efflux rate increased in the sequence: N1-methylnicotinamide (NMeN+) < cimetidine < tetraethylammonium (TEA+) < N-methyl-4-phenylpyridinium (MPP+). Preloading the animals by i.v. infusion or pre perfusion of the peritubular capillaries with NMeN+ increased the efflux rate of MPP+. Luminal efflux was also augmented when the tubular solution was made alkaline with HCO3- or phosphate, whereby HCO3- is more effective than phosphate. Replacement of Na+ by Cs+ showed no effect. With i.v. preloading the animals with NMeN+ and with 25 mM HCO3- in the luminal perfusate the 2-s efflux follows kinetics with a Michaelis constant Km = 0.21 mmol/l and maximal flux Jmax = 0.42 pmol.cm-1.s-1 and a permeability term with P = 37.7 microns2.s-1. Comparing the apparent luminal inhibitory constant values for MPP+ (Kil,MPP+) with the apparent contraluminal Kicl,NMeN+ values of substrates of homologous series, it was found that (1) limitation by molecular size occurs at the contraluminal cell side earlier than at the luminal cell side; (2) affinity increases with hydrophobicity of the substrates at the luminal cell side, with a steeper or equal slope than at the contraluminal cell side; (3) affinity increases with basicity (i.e. pKa values) at the luminal cell side with a steeper slope than at the contraluminal cell side. Taken together, substrates with low hydrophobicity and low basicity interact at the luminal cell side more weakly than at the contraluminal cell side. On the other hand large, hydrophobic substrates have, at the luminal cell side, a higher affinity than at the contraluminal cell side. Many substrates, however, have equal affinity at the luminal and contraluminal cell sides. PMID- 7491275 TI - Effect of dexamethasone on sodium channel block and densities in A6 cells. AB - The association (ON) and dissociation (OFF) rates of either positively charged amiloride or its uncharged analogue, CDPC (6-chloro-3, 5-diaminopyrazine-2 carboxamide), with the apical Na+ channel protein of renal A6 cells were analysed during exposure to the synthetic glucocorticoid, dexamethasone, using noise analysis. These rates were further used to reach specific conclusions about single-channel current, channel density and open probability of the channel in the absence of the blocker. Short-term exposure (3 h) to 10(-7) mol/l dexamethasone at the basolateral side increased the short-circuit current, Isc by 85%, without a change in the ON and OFF rates of the interaction between amiloride and the Na+ channel. A longer incubation (24 h) with dexamethasone tripled the current with a notable increase in the ON rate of the interaction between amiloride and the and channel. The OFF rate remained constant. The effects of dexamethasone on the rate constants of the reaction of amiloride with the channel did not match with the expected changes in membrane potential. On the other hand, ON and OFF rates of the interaction between neutral CDPC and the channel were not influenced by a 24-h incubation with dexamethasone. Further calculations disclosed that the gain in macroscopic current after a 24-h incubation with dexamethasone might be explained by an increase in Na+ channel density, and, to a lesser extent, by a rise in single-channel current. This all occurred without a change in the fraction of time spent by the channel in the conducting state in the absence of the blocker. PMID- 7491278 TI - Diffusion of myoglobin in skeletal muscle cells--dependence on fibre type, contraction and temperature. AB - We measured the diffusion coefficient of myoglobin (DMb) inside mammalian skeletal muscle cells with a microinjection technique. A small bolus of horse Mb was injected into a single muscle fibre and the subsequent time-dependent changes of the Mb profiles along the fibre axis were measured with a microscope photometer. For fibres of the rat soleus muscle at 22 degrees C, a DMb of 1.3.10( 7) cm2/s was found, confirming a result obtained previously by us for rat diaphragm muscle with a photo-oxidation technique. In the extensor digitorum longus muscle of the rat, a higher value of 1.9.10(-7) cm2/s was measured. Auxotonic muscle contractions did not change the apparent DMb. For the temperature range between 22 degrees C and 37 degrees C, a temperature coefficient. Q10, of 1.5 was calculated. The implication of this result for the role of Mb in the facilitation of oxygen transport was examined. Model calculations show that with this relatively low DMb value, the intracellular oxygen supply can be improved only slightly. PMID- 7491276 TI - Effects of polyamines on voltage-activated calcium channels in guinea-pig intestinal smooth muscle. AB - Effects of polyamines on the spontaneous mechanical and electrical activity of guinea-pig intestinal smooth muscle were studied. Spermine and spermidine inhibited action potential generation and contractions, while putrescine had no effect. Single smooth muscle cells were isolated from the longitudinal muscle layer of the guinea-pig ileum. Whole-cell voltage-clamp experiments were carried out to investigate the effects of polyamines on current through voltage-activated Ca2+ channels. Spermine and spermidine (0.1-1 mM) reduced the inward current in a concentration-dependent manner. Spermine blocked current activated by the dihydropyridine agonist BAY K 8644 (1 microM), whereas no additional inhibition by spermine was seen after blockage of dihydropyridine-sensitive channels by nifedipine (0.1 microM). Inhibition by spermine or spermidine did not shift the peak of the current voltage relation of the inward current. Steady-state activation and inactivation relationships were not affected and thus the amplitude, but not the voltage dependence, of the window current responsible for Ca2+ inflow during sustained depolarization was affected. Putrescine (1 mM) had no significant effect on the inward current. These results suggest that spermine and spermidine inhibit contraction in spontaneously active intestinal smooth muscle by inhibiting Ca2+ current responsible for generation of action potentials. PMID- 7491277 TI - Optical multisite monitoring of cell excitation phenomena in isolated cardiomyocytes. AB - An especially designed setup which consists of an inverted fluorescence microscope, an argon ion laser and a photodiode array system permits membrane potential monitoring in isolated guinea-pig ventricular cardiomyocytes, stained with the voltage-sensitive dye di-4-ANEPPS, which responds linearly with relative fluorescence changes (delta F/F) approximately -8% per 100 mV. About a dozen measuring spots covering a single cell were simultaneously monitored with a spatial and temporal resolution of 15 microns and about 20 microseconds, respectively. In general, the rising phases of the action potentials within a single cell were highly synchronized (i.e. all upstroke velocities peaked within about 20 microseconds); however, in one cell (out of 25 examined) significant (P < 0.05) time lags exceeding the signal-dependent time resolution were also found. Experiments, simultaneously performed with our optical system and a widely used patch-clamp setup, revealed a slowed and delayed response of the clamp amplifier depending on the cell access resistance. Optical monitoring during whole-cell voltage-clamping demonstrated the influence of graduated series resistance compensation. When field stimulation was used, our results clearly demonstrated the spatially dependent polarization of the cell membrane during the stimulus, as well as a highly synchronized upstroke development. Slight differences in the maximum upstroke velocities within a single cell were also found and were basically in agreement with mathematical models. PMID- 7491280 TI - Neuropeptide Y2-type receptor-mediated activation of large-conductance Ca(2+) sensitive K+ channels in a human neuroblastoma cell line. AB - We have proposed recently that a pertussistoxin-insensitive Ca2+ influx stimulated by Y2-type receptor activation in CHP-234 human neuroblastoma cells underlies increases in intracellular free Ca2+ concentration ([Ca2+]i) induced by neuropeptide Y (NPY), which were strictly dependent on extracellular Ca2+ and independent of internal Ca2+ stores. We describe here the actions of NPY in these same cells, using the activity of Ca(2+)-activated K+ channels as an indicator of [Ca2+]i. The elementary slope conductance of these channels was 110 +/- 3 pS (with an asymmetrical K+ gradient), their activity was greatly increased by application of ionomycin, and they were reversibly blocked by 1 mM tetraethylammonium (TEA) and 100 nM charybdotoxin. Application of 100 nM NPY, in the presence but not in the absence of extracellular Ca2+, increased the channel open probability. ATP applied in the absence of external Ca2+ caused rises both in channel open probability and [Ca2+]i. Inositol trisphosphate production was stimulated by ATP but not by NPY. In outside-out patches, NPY increased channel open probability, indicating that NPY-associated Ca2+ influx does not require all the intracellular machinery present in intact cells. Channel activation by NPY was unaffected by the replacement of guanosine 5'-triphosphate (GTP) by (guanosine 5'-O-(2-thiodiphosphate) (GDP[ beta S]), a non-hydrolysable GDP analogue, in the pipette internal solution, consistent with the lack of involvement of G-proteins in the coupling of Y2-type receptors to Ca2+ influx in CHP-234 cells. PMID- 7491279 TI - Properties of voltage-gated currents of microglia developed using macrophage colony-stimulating factor. AB - Microglia were isolated from a murine neonatal brain cell culture in which their development had been stimulated by supplementation with the macrophage/microglial growth factor macrophage colony-stimulating factor (M-CSF). Using the whole-cell configuration of the patch-clamp technique, voltage-gated membrane currents were recorded from these microglial cells. Hyperpolarization induced inward rectifying K+ currents, as described for microglia from untreated cultures. These currents activated negative to the K+ equilibrium potential and, with a strong hyperpolarization, displayed time-dependent inactivation. The inactivation was abolished when extracellular NaCl was replaced by N-methyl-D-glucamine (NMG), thereby indicating a partial block of this K+ conductance by Na+. Inward rectifying currents were also blocked by extracellularly applied Cs+ or Ba2+. They were slightly diminished following treatment with extracellular tetraethylammonium chloride (TEA) but were not affected by 4-aminopyridine (4 AP). Upon long lasting depolarizing voltage pulses to potentials positive to 0 mV, the cells exhibited a slowly activating H+ current which could be reduced by application of inorganic polyvalent cations (Ba2+, Cd2+, Co2+, La3+, Ni2+, Zn2+) as well as by 4-AP or TEA. Based on their kinetics and pharmacological characteristics, both currents detected on M-CSF-grown microglia are suggested to correspond to the inward rectifier and the H+ current of macrophages. PMID- 7491281 TI - Fast, persistent, Ca(2+)-dependent K+ current controls graded electrical activity in crayfish muscle. AB - The early outward current in opener muscle fibres of crayfish (Procambarus clarkii) was studied using the two-electrode voltage-clamp technique. This current was abolished in Ca(2+)-free and 5 mM Cd2+ solutions, and was blocked by extra- or intracellular tetraethylammonium, indicating that it was a Ca(2+) dependent K+ current [IK(Ca)]. IK(Ca) was voltage dependent, apamin insensitive and sensitive to charybdotoxin (CTX), which, in addition to its tetraethylammonium sensitivity, suggests that the channels mediating IK(Ca) behave in a BK type manner. IK(Ca) activation was extremely fast, reaching a maximum within 5 ms, and the inactivation was incomplete, stabilizing at a persistent steady-state. IK(Ca) was insensitive to intracellular ethylenebis(oxonitrilo)tetraacetate (EGTA), but was abolished by injection of the faster Ca2+ chelator 1,2-bis(2-aminophenoxy)ethane-N,N,N',N'-tetraacetic acid (BAPTA), suggesting that voltage-dependent Ca2+ channels and those mediating IK(Ca) should be clustered closely on the membrane. Under two-electrode current clamp recording mode, low amplitude, graded responses were evoked under control conditions, whereas repetitive all-or-none spikes were elicited by application of CTX or after loading the cells with BAPTA. We conclude that IK(Ca) activates extremely quickly, is persistent and is responsible for the generation and control of the low amplitude, graded, active responses of opener muscle fibres. PMID- 7491282 TI - Ionic basis of neurokinin-A-induced depolarization in single smooth muscle cells isolated from guinea-pig trachea. AB - Neurokinin A (NKA) caused single tracheal smooth muscle cells (TSMCs) to contract. The effects of NKA on the electrical activity of guinea-pig TSMCs were examined using the tight-seal whole-cell patch-clamp technique. Under current clamp conditions at rest, the membrane potential of TSMCs spontaneously oscillated at about -40 mV and NKA rapidly depolarized the membrane potential to nearly 0 mV, which then gradually repolarized to about -20 mV in the presence of NKA. The oscillations in potential disappeared transiently during the rapid phase of depolarization in response to NKA and reappeared during the sustained phase of depolarization. Under voltage-clamp conditions, NKA evoked an inward current which faded quickly. Subsequently, the cell conductance in the presence of NKA at potentials greater than -40 mV decreased gradually. The reversal potential of the NKA-induced inward current was about 0 mV, and shifted with changes in the Cl- equilibrium potential. The Cl- current was not elicited by NKA when using a pipette solution containing 10 mM ethylenebis(oxonitrilo)tetraacetic acid (EGTA). During the sustained phase, K+ currents evoked by depolarizing voltage steps were inhibited by NKA. The present results indicate that NKA causes rapid and sustained depolarization of TSMCs by two distinct mechanisms: (1) initial transient activation of the Ca(2+)-dependent Cl- current, and (2) sustained inhibition of K+ currents. PMID- 7491283 TI - Potassium currents in rat colonic smooth muscle cells and changes during development and aging. AB - In a previous study on freshly isolated single smooth muscle cells from the circular layer of the rat distal colon, we reported that the L-type Ca2+ current density increased during development and gradually declined with further aging [ZI Xiong, N Sperelakis, N Noffsinger, C Fenoglio-Preiser (1993) Am J Physiol 265: C617-C625]. Since K+ current plays a key role in controlling excitability of the cells and hence the motility of the colon, in the present study the voltage gated K+ channel currents, (IK) were investigated using the whole-cell voltage clamp technique in colonic myocytes from rats of different ages. A Ca(2+) sensitive K+ current [IK(Ca)] and two kinds of Ca(2+)-insensitive outward K+ currents were identified and characterized. IK(Ca) was recorded at potentials more positive than -40 mV in Ca(2+)-containing bath solution, and was blocked by Ca2+ channel antagonists and tetraethylammonium ion (TEA+). After removing Ca2+ from the bath solution and using a high ethylenebis(oxonitrilo)tetraacetate (EGTA, 4 mM) concentration in the pipette, two types of Ca(2+)-insensitive IK were recorded. The first and faster component was usually activated at potentials more positive than -50 mV, and was more sensitive to 4-aminopyridine (4-AP). In contrast, the second and slower (delayed) component was activated at potentials more positive than -30 mV, and was more sensitive to TEA. The total density of the Ca(2+)-insensitive IK component decreased dramatically during the neonatal period: from 32.2 +/- 3.2 pA/pF in 3-day-old rats to 17.8 +/- 2.6 pA/pF in 40-day old rats; there was no further decline during aging (up to 480 days).(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7491284 TI - Relaxation in ferret ventricular myocytes: role of the sarcolemmal Ca ATPase. AB - In ferret ventricular myocytes the rate of intracellular Ca concentration [Ca]i decline and relaxation is remarkably fast (compared with rabbit and rat) under conditions where both the sarcoplasmic reticulum Ca uptake and Na/Ca exchange are inhibited. Here we explore the possibility that this rapid [Ca]i decline in ferret cells is attributable to the sarcolemmal Ca ATPase by using carboxyeosin (a potent inhibitor of the sarcolemmal Ca-ATPase). We compare the effects of carboxyeosin with those of elevated extracellular [Ca] ([Ca]o) (a thermodynamic approach to limit Ca transport by the sarcolemmal Ca ATPase). In rabbit cells, carboxyeosin and high [Ca]o slowed [Ca]i decline similarly and both virtually abolished [Ca]i decline when mitochondrial Ca uptake was also inhibited. In ferret cells, carboxyeosin treatment produced these same effects on [Ca]i decline, but high [Ca]o did not mimic them. Moreover, only in carboxyeosin treated ferret cells did additional inhibition of mitochondrial Ca uptake nearly abolish [Ca]i decline. We conclude that, carboxyeosin loading can inhibit the sarcolemmal Ca-ATPase in intact myocytes; that this pump seems likely to be responsible for the much faster relaxation observed in ferret cells after block of SR Ca accumulation and Na/Ca exchange transport and that the sarcolemmal Ca pump apparently has different characteristics in rabbit and ferret ventricular myocytes. PMID- 7491285 TI - A method for recording intracellular [Ca2+] transients in cardiac myocytes using calcium green-2. AB - Calcium green-2 (Ca green) is a non-ratiometric fluorescent Ca2+ indicator with an affinity for Ca2+ (dissociation constant Kd = 3 microM) that is lower than more commonly used indicators such as fura-2 and fluo-3. This low Ca2+ affinity, coupled with a high quantum yield, allows cells to be loaded with low concentrations of Ca green, avoiding problems of cytosolic Ca2+ buffering and a low signal-to-noise ratio. This communication presents a method for monitoring intracellular [Ca2+] changes in isolated rat ventricular myocytes loaded with Ca green and the fluorescent pH indicator carboxy SNARF-1 (SNARF). SNARF provides a Ca(2+)-insensitive signal with which Ca green fluorescence can be corrected for cell motion and dye-loading artifacts. PMID- 7491286 TI - A novel method for incorporation of ion channels into a planar phospholipid bilayer which allows solution changes on a millisecond timescale. AB - We have developed a method of rapidly changing the solutions on one side of a planar phospholipid bilayer. Bilayers can be painted on glass pipettes of tip diameter > or = 50 microns. By modifying an established method for rapid exchange of solutions bathing excised membrane patches, solution changes can be made at the bilayer within 10 ms. After incorporation of channels into the bilayer, the bilayer is moved into one of two parallel streams of solution flowing from a length of double-barrelled glass theta tubing. Activation of a solenoid system rapidly moves the theta tubing so that the bilayer is in the flow of the adjacent solution. For various reasons, the single-channel gating mechanisms of many channels are studied in planar bilayer systems. The conventional bilayer technique only allows for steady-state single-channel gating to be monitored. This novel method now allows the effects of rapid changes in modulators of channels incorporated into planar phospholipid bilayers to be measured. PMID- 7491290 TI - Extracellular protons modulate the Ca2+ block of a Ca(2+)-blockable monovalent cation channel in chick embryo. AB - Some epithelial nonselective cation channels although in an open conformation are impermeable because they are blocked by extracellular Ca2+ ions. Such a channel is present in the apical membrane of the ectodermal cells of the chick gastrula. This single-channel study shows that protonation of a site located inside the pore of this embryonic channel reduces the Ca2+ block thus allowing Na+ to pass. The apparent pK for Ca2+ at -50 mV shifts from 5.7 at pH 7.4 to 3.5 at pH 4.5. Unexpectedly, the change in Ca2+ affinity does not originate from a destabilization of the binding but comes from a large reduction of the association rate. PMID- 7491289 TI - Tetrodotoxin-sensitive fast Na+ current in embryonic chicken osteoclasts. AB - A voltage-dependent, fast, transient inward current was characterized in embryonic chicken osteoclasts using the permeabilized patch configuration of the patch-clamp technique. The current was activated by depolarizations to higher than -28 +/- 4 mV from a holding potential of -80 mV. It peaked within 1-1.5 ms, and inactivated within 3.3-6.9 ms. The 50% inactivation voltage was -59 +/- 6 mV with a steepness factor of 0.11 +/- 0.06. The current disappeared with the removal of extracellular Na+ and was reversibly blocked by tetrodotoxin (K0.5 < 15 nM) but not by verapamil (< or = 100 microM). We conclude that this new current in embryonic chicken osteoclasts is a sodium current known from excitable cells. PMID- 7491287 TI - A Gi1-2-protein is required for alpha 2A-adrenoceptor-induced stimulation of voltage-dependent Ca2+ channels in rat portal vein myocytes. AB - In rat portal vein myocytes, alpha 2A-adrenoceptors activate voltage-dependent Ca2+ channels via a transduction pathway requiring protein kinase C activation mediated by a pertussis-toxin-sensitive G-protein. As revealed by the use of antibodies directed against the different alpha-subunits expressed in portal vein myocytes, we show that the clonidine-induced stimulation of voltage-dependent Ca2+ channels is mainly mediated by a Gi1-2-protein. PMID- 7491288 TI - Coupling of exocytosis to depolarization in rat pancreatic islet beta-cells: effects of Ca2+, Sr2+ and Ba(2+)-containing extracellular solutions. AB - Using rat beta-cells we present evidence that Sr2+ and Ba2+, like Ca2+, support depolarization-induced increases in membrane capacitance which reflect insulin granule exocytosis. Even with identical total charge entry, Sr2+ and Ba2+ are 3-5 and 20-fold less effective than Ca2+ in supporting release. While exocytosis supported by Sr2+ is graded with cation entry and complete within 250ms of depolarization, exocytosis supported by Ba2+ begins abruptly after a threshold of charge entry and continues for many seconds. Ba(2+)-supported release continues in the presence of greatly enhanced cytosolic Ca2+ buffering, arguing against release of Ca2+ from stores as its principal action. These results suggest that Sr2+ and Ba2+ support exocytosis largely by binding to Ca(2+)-dependent release activating sites, though with less affinity than Ca2+. PMID- 7491291 TI - Distribution of intracellular calcium in isolated mouse skeletal muscle fibres. PMID- 7491292 TI - [The 31st Autumn meeting of the Japan Radiological Society. October 23-25, Fukui prefecture, Japan. Abstracts]. PMID- 7491296 TI - Effects of linoleic acid and gamma-linolenic acid on the growth and metastasis of a human breast cancer cell line in nude mice and on its growth and invasive capacity in vitro. AB - It has been reported that gamma-linolenic acid (GLA)-rich diets suppress mammary carcinogenesis and transplanted tumor growth and that GLA inhibits the growth of cultured human cancer cell lines. We compared the effects of dietary GLA and linoleic acid (LA) on the growth of MDA-MB-435 human breast cancer cells and their expression of the metastatic phenotype in vivo and in vitro. Athymic nude mice (30/dietary group) were fed isocaloric diets containing 20% (wt/wt) fat but providing 8% GLA or LA for 7 days, and 10(6) tumor cells were then injected into a thoracic mammary fat pad. The diets were continued for a further 11 weeks. The primary tumor growth rates were similar in mice from the two dietary groups; there was a nonstatistically significant trend for the incidence of macroscopic lung metastases and the total lung metastatic volumes to be higher in the GLA-fed mice (79% and 40.1 +/- 13.9 mm3) than in the LA-fed mice (64% and 15.5 +/- 5.4 mm3). The tumor cell phospholipids from the 8% GLA-fed mice contained significantly lower LA levels but higher arachidonic acid levels (both p < 0.001) than those from 8% LA-fed mice. Also the arachidonate-derived eicosanoids (prostaglandin E, leukotriene B4, and 5-, 12-, and 15-hydroxyeicosatetraenoic acids) were significantly higher in tumors from the 8% GLA group. Zymography showed higher 92-kDa type IV collagenase activity in tumors from 8% GLA-fed mice. In vitro, GLA and LA, at 0.5-2 micrograms/ml, stimulated MDA-MB-435 cell growth; 10 micrograms/ml was mildly inhibitory. Whereas LA stimulated tumor cell invasion and 92-kDa type IV collagenase production in vitro, GLA inhibited invasion and did not induce activity of the proteolytic enzyme. Our results do not support the hypothesis that supplementation with GLA would exert a beneficial effect on the progression of an existing breast cancer, perhaps because it is metabolized in vivo to arachidonate-derived eicosanoids that are known to be involved in the metastatic process. PMID- 7491294 TI - Dietary fat: gene expression and mammary tumorigenesis. AB - Although discovered as an exogenous agent of mammary carcinoma, the mouse mammary tumor virus (MMTV) is also transmitted vertically as endogenous proviral DNA present in the germ line of all inbred mice. In the C3Hf mouse, which receives no exogenous virus, the initial event of mammary tumorigenesis is the transcription of the endogenous MMTV proviral DNA present at the Mtv-1 locus. Transcription occurs as a result of the hormonal effects of pregnancy, and Mtv-1 specific transcripts are seen in second-parity lactating mammary glands of these mice. As a means of studying the effects of diet on mammary carcinoma at the molecular/genetic level, we have studied the transcription of the Mtv-1 locus in C3Hf mice on a high-fat diet containing 46% fat in calories or a low-fat diet containing 10% fat in calories. We have detected an accelerated transcription of the Mtv-1 locus (first- vs. second-parity lactating mammary glands) in > 50% of the C3Hf mice on the high-fat diet. In addition, mice on the high-fat diet developed mammary tumors earlier (11 vs. 17.8 mos) and after fewer litters (2.1 vs. 4.2). Our results indicate that fat in the diet can affect gene expression related to mammary carcinoma. PMID- 7491295 TI - Accumulation of beta-carotene in normal colorectal mucosa and colonic neoplastic lesions in humans. AB - The quantity of beta-carotene (BC) accumulated in colonic polyps and colonic cancerous tissue in humans in situ was determined relative to the quantity accumulated in normal colon and rectal tissue. Serum concentration of BC, retinol, and alpha-tocopherol and tissue BC concentration were determined by high performance liquid chromatography in samples obtained before and after oral supplementation with BC (30 mg/day). The serum BC and retinol concentrations significantly increased in response to supplementation in control, polyp, and cancer patients, but there was no change in serum alpha-tocopherol concentration. The BC concentration in tissue (colon, rectum, and tumor) of cancer patients was significantly less than that in tissue samples from control and polyp patients. Relative to baseline values, BC accumulated to a significant extent in tissues from all patients, including polyp and tumor tissue, during supplementation. The results indicate that BC does accumulate in colonic neoplastic tissue in humans and may potentially be utilized to augment cytotoxicity of chemotherapeutics or to prevent malignant transformation of cells. PMID- 7491293 TI - Urinary lignan and isoflavonoid excretion in men and women consuming vegetable and soy diets. AB - Urinary lignan and isoflavonoid excretion were examined in 11 men and 9 women consuming four nine-day controlled experimental diets: basal (vegetable free), carotenoid vegetable (carrot and spinach), cruciferous vegetable (broccoli and cauliflower), and soy (tofu and textured vegetable protein product). Three-day urine collections (Days 7-9) were analyzed for lignans and isoflavonoids with use of isotope-dilution gas chromatography-mass spectrometry. Urinary excretion of the lignans enterodiol and enterolactone was higher during the carotenoid and cruciferous vegetable diets than during the basal diet (p = 0.0001), suggesting that these vegetables may provide a source of mammalian lignan precursors. Urinary excretion of the isoflavonoids equol, O-desmethylangolensin, daidzein, and genistein was higher when subjects consumed soy diets than when they consumed the other test diets (p < 0.02). Gender differences in lignan excretion were observed. Men excreted more enterolactone (p = 0.006) and less enterodiol (p = 0.013) than women, implying a gender difference in colonic bacterial metabolism of lignans. There was no effect of gender on isoflavonoid excretion. PMID- 7491297 TI - Altered cytokeratin expression in carcinogenesis inhibition by antioxidant nutrients. AB - Epidermoid carcinomas were induced in hamster buccal pouches with use of 7.12 dimethylbenz[a]anthracene (DMBA). In five animals that served as tumor controls (Group 1), right buccal pouches were painted with DMBA (0.5% solution in mineral oil) thrice weekly for 14 weeks. In five animals (Group 2), right buccal pouches were painted with DMBA and reduced glutathione (GSH) was administered systemically by mouth. Five animals (Group 3) received vitamin E instead of glutathione. An additional 20 animals (Groups 4, 5, 6, and 7) were untreated, vehicle, glutathione, and vitamin E controls, respectively. Glutathione and vitamin E were given in doses of 10 mg/kg in 0.5 ml of mineral oil thrice weekly on days alternate to DMBA painting. Treatment by GSH and vitamin E reduced the number and size of tumors that were formed. Histopathologically, there were also fewer sites of dysplasia, carcinoma in situ, and early invasive epidermoid carcinoma than in the tumor control animals. The formalin-fixed and paraffin embedded buccal pouch sections were stained immunohistochemically with use of monoclonal antibodies for cytokeratins. These included high-molecular-weight keratins (50,000-68,000 mol wt) 10, 13, and 8 (k10, k13, and k8, respectively). Oral carcinomas and dysplastic sites exhibited basal and suprabasal (spinous layer) high levels of k10, k13, and k8 staining. Treatment with GSH or vitamin E increased the suprabasal staining for high-molecular-weight keratins and reduced the protein expression for k10, k13, or k8. This pattern of staining was observed in dysplastic as well as in carcinoma sites. These results indicate that cytokeratin protein expression could contribute to a common biomarker analysis for chemoprevention. PMID- 7491299 TI - Influence of starch fermentation on bile acid metabolism by colonic bacteria. AB - Secondary bile acids (BA) may be involved in the pathogenesis of colorectal cancer. In vivo, starch malabsorption has been shown to reduce fecal excretion of secondary BA. The present in vitro study was performed to investigate the effect of starch fermentation on BA metabolism by colonic bacteria. Fecal samples of healthy volunteers were incubated in anaerobic batch cultures for 48 hours with the primary bile acids cholic (0.6 g/l) and chenodeoxycholic acid (0.4 g/l). Media were starch free or enriched with starch (10 g/l). The pH was controlled and held at 6 or 7. In the starch-free incubations, secondary BA were rapidly formed, and degradation of primary to secondary BA was complete within 24 hours. The formation of secondary BA was partially inhibited by the addition of starch to the media. This effect was stronger at pH 6 than at pH 7. Starch was rapidly and completely fermented. In conclusion, this study showed that formation of secondary BA by fecal bacteria is inhibited when starch is simultaneously fermented, an effect that is mainly, but not completely, explained by reduction of pH. PMID- 7491298 TI - Unsaturated fatty acid effects on human breast cancer cell adhesion. AB - Polyunsaturated fatty acids influence several steps involved in metastasis formation in animal tumor models. During the process of metastasis from the primary site, tumor cells adhere to the endothelium and underlying basement membrane before extravasation and secondary growth. The purpose of this study was to determine the effect of unsaturated fatty acids on adhesion of human breast cancer cell lines to components of the basement membrane. Cells were cultured in low-serum medium for five days with or without added unsaturated fatty acids. Adhesion assays were conducted by incubating cells with basement membrane substrates coated on 96-well plates, washing to remove nonadherent cells, and staining adherent cells with crystal violet. Linoleic acid (LA) and eicosapentaenoic acid increased adhesion of the metastatic cell line MDA-MB-231 to Matrigel and type IV collagen, while eicosapentaenoic acid decreased adhesion of the less metastatic cell line SK-BR-3 to these two basement membrane substrates. Oleic acid increased adhesion of MDA-MB-231 cells to Matrigel and fibronectin. Nordihydroguaiaretic acid and high concentrations of indomethacin, each of which inhibits the lipoxygenase pathway of arachidonate metabolism, were effective in reversing the stimulatory effect of LA on MDA-MB-231 cell adhesion. A protein kinase C inhibitor likewise suppressed the increase in adhesion observed when MDA-MB-231 cells were incubated in media with added LA. Unsaturated fatty acids modified the adhesive properties of human breast cancer cell lines in vitro, and LA appeared to increase human breast cancer cell adhesion to extracellular matrix components by activating lipoxygenase and/or protein kinase C pathways. PMID- 7491300 TI - Developing Yoshida sarcoma does not influence the fatty acid composition of adipose tissues in the rat. AB - The potential involvement of individual fatty acids from diet or from adipose tissue in the outcome of cancer emphasizes the need for more insight into the relationship between fatty acids and tumor growth. The main objective of the present study was to examine whether rapid tumor growth would induce changes in the fatty acid composition of adipose tissue, indicating selective use and thus deposition of dietary fatty acids and/or selective removal of stored fatty acids from adipose tissue. We used a rodent model of transplanted solid tumor (Yoshida sarcoma) and measured the fatty acid composition from different adipose sites in the absence and in the presence of tumor but at the same intake of dietary fatty acids. We found that Yoshida sarcoma could grow rapidly without significantly altering the fatty acid composition of adipose tissue, irrespective of its location. These results do not favor the hypothesis of a selective removal of fatty acids from adipose tissue by growing tumors. They moreover indicate that tumors do not modify the storage of individual dietary fatty acids. PMID- 7491301 TI - Reproducibility of food intake in a food frequency questionnaire used in a general population. AB - This study evaluates the reproducibility of food intake reports in a 45-item self administered food frequency questionnaire (FFQ) used for a diet study in a large cohort in New York State. The first FFQ was mailed in 1980, and a followup version containing 75 food items was mailed in 1988 to the eligible original participants. Reproducibility was evaluated by comparing the subjects' 1988 responses with their original 1980 responses. Spearman's correlations ranged from 0.27 (nuts other than peanuts) to 0.56 (summer squash) in men and from 0.25 (tomatoes) to 0.51 (summer squash) in women. Analyses stratified by age, education, marital status, body mass index, cigarette smoking, alcohol drinking, and special diet revealed few substantial differences in correlations. The moderate reproducibility of foods in this questionnaire indicates that brief FFQs may be a useful tool to study food intake and chronic disease relationships in this population. PMID- 7491302 TI - Nonthoracotomy ICD implantation: lessons to be learned from permanent pacemaker implantation. PMID- 7491303 TI - Atrial pacing leads: the clinical contribution of steroid elution. AB - Although the original atrial pacing leads were passive fixation and J shaped for right atrial appendage placement, the subsequent development of the active fixation screw-in lead found favor because of a perceived low incidence of lead dislodgment and a wider selection of atrial pacing sites. A bipolar atrial lead study was undertaken to compare the long-term atrial implant data in 215 patients. Study leads comprised one passive fixation, steroid-eluting lead (Medtronic CapSure SP, 119 patients) and three nonsteroid-eluting leads; two active fixation (Medtronic BISPING model 4058, 30 patients; and Telectronics ACCUFIX model 330-801, 44 patients) and one passive fixation (Telectronics ENCOR model 330-854, 22 patients). Bipolar atrial voltage stimulation thresholds and electrograms were measured using the Telectronics META DDDR immediately postimplantation, and at 1-, 3-, 6-, 12-, and 18-month follow-up. There were 135 males and the mean age 68 years. The incidence of lead dislodgment was 4% for active fixation and 2% for passive fixation. All nonsteroid leads showed a typical rise in stimulation threshold with the highest being the ACCUFIX followed by the BISPING and ENCOR. The steroid-eluting CapSure SP, however, demonstrated a flat response with 98% of leads at 18 months having a value < or = 1.3 volts allowing voltage programming to 2.5 volts (2:1 safety ratio). Telemeted electrograms showed no differences for all leads at all visits. For low voltage atrial pacing with a low incidence of dislodgment and satisfactory atrial sensing, the steroid-eluting passive fixation lead is superior to all nonsteroid eluting leads. PMID- 7491304 TI - Long-term follow-up of patients treated by radiofrequency ablation of the atrioventricular junction. AB - Radiofrequency ablation of the AV conduction tissue (His-bundle ablation) is an accepted treatment for therapy resistant atrial fibrillation/flutter. However, data on the long-term effects of the procedure are limited. We followed 50 patients for a mean of 17 months after AV junction ablation. The indication was treatment resistant atrial fibrillation or flutter. The patients underwent a standardized interview performed by two nurses. Health care was studied via the in-patient register. Subjective improvement was reported by 88% and the number of days in hospital per year was reduced from 17 to 7. The use of antiarrhythmic drugs was reduced by 75%. If the reduction in costs of drugs and days in hospital is compared with the cost of the ablation and the pacemaker implantation, breaking even is achieved after 2.6 years. We could not confirm that patients with paroxysmal atrial fibrillation note less improvement than those with chronic fibrillation. CONCLUSION: Ablation of the AV junction is a cost effective treatment with good long-term results and relatively few complications. RECOMMENDATIONS: Chronic atrial fibrillation: If sinus rhythm cannot be established and in cases in which heart rate regulating drugs have been ineffective, ablation of the AV junction with implantation of a VVIR pacemaker is recommended. PAROXYSMAL ATRIAL FIBRILLATION: If the patient despite treatment with antiarrhythmic drugs continues to have symptomatic episodes of atrial fibrillation, then AV junction ablation with implantation of a permanent pacemaker is recommended. Patients who have self-limiting episodes of atrial fibrillation should be given a DDDR pacemaker with an automatic mode switch. Patients who do not have self-limiting attacks and require DC conversion, should receive a VVIR pacemaker. PMID- 7491305 TI - A fast and reliable algorithm to localize accessory pathways based on the polarity of the QRS complex on the surface ECG during sinus rhythm. AB - BACKGROUND: Many criteria have been published to localize accessory pathways from the 12-lead ECG during sinus rhythm. This study analyzed whether the localization of an accessory pathway could be predicted by using the polarity of the QRS complex during sinus rhythm on the surface ECG, instead of the delta wave polarity as used in many reports. METHODS: The ECGs of 140 patients with an overt and single accessory pathway were evaluated. Eight localizations were taken into account. The precise location was previously known from successful radiofrequency ablation sites. RESULTS: In 128 patients (92%), the new algorithm allowed an accurate diagnosis of the site of implantation of the accessory pathway. CONCLUSION: Analysis of the polarity of the QRS complex on five electrocardiographic leads provides an easy, fast and reliable way to localize accessory pathways during sinus rhythm. PMID- 7491306 TI - Effects of atrial, ventricular, and atrioventricular sequential pacing on coronary flow reserve. AB - Experimental animal data have indicated that altered left ventricular depolarization sequence as a result of right ventricular pacing may diminish coronary blood flow in the distribution of the left anterior descending coronary artery. To further investigate this, we compared the effects of atrial, ventricular, and atrioventricular (AV) sequential pacing on coronary flow reserve. Twenty-seven patients (24 male, mean age 55 +/- 7 years) with normal left anterior descending coronary arteries were studied. Coronary flow reserve was calculated as the ratio of mean flow velocity at maximal coronary vasodilatation to mean flow velocity at baseline. The study consisted of two parts. In the first part, AV sequential pacing was compared to atrial pacing at the same rate; coronary flow reserve did not differ significantly between the two pacing modes (14 patients, 4.85 +/- 1.88 vs 5.47 +/- 1.55, respectively, P > 0.05). In the second part, all three pacing modalities were compared; coronary flow reserve was significantly higher during ventricular compared to AV sequential pacing, but not significantly different compared to atrial pacing (3.69 +/- 1.42 vs 2.90 +/- 0.86 vs 3.11 +/- 0.89, respectively, P < 0.05). This difference was secondary to a significant decrease in mean baseline velocity during ventricular pacing, while mean velocity during hyperemia was comparable between the three pacing modes. It is concluded that AV sequential pacing does not appear to exert a significant effect on coronary flow reserve. Ventricular pacing, however, may lower resting coronary blood velocity in some patients, without affecting maximal coronary blood velocity, resulting in a higher coronary flow reserve. PMID- 7491307 TI - Preliminary evaluation of a dual chamber pacemaker with bradycardia diagnostic functions. AB - Unexplained syncope is the main indication for the implantation of a diagnostic pacemaker. Studies on those implanted have shown that in patients with unexplained syncope, the diagnosis of paroxysmal bradycardia was feasible and reliable. The present study was designed to evaluate a new bradycardia diagnosis algorithm, loaded in a dual chamber pacemaker, in 24 patients considered as candidates for diagnostic pacemakers. During a mean follow-up of 153 days, at least one bradycardia episode was recorded in 13 patients. The mean number of detected bradycardias was 6 and the median was 3. The mean delay between the algorithm activation and the first bradycardia episode was 67 days. The mechanism of bradycardia was atrioventricular block in 6 patients, sinus node dysfunction in 6 patients, and consecutive blocked atrial premature beats in 1 patient, as indicated by the event markers. In 11 patients bradycardia was recorded during the daytime only or day and night. In two patients the episodes were recorded only at night. Overall, the algorithm was well-tolerated; however, some mild symptoms were observed due to the method of bradycardia determination, allowing bradycardia. Three patients were symptomatic as a direct result of the algorithm operation, and four patients had symptoms related to the single chamber operation of the pacemaker while functioning in the diagnosis mode (VDI). These symptoms were relieved with DDD pacing. PMID- 7491308 TI - Effects of electroporation on transmembrane potential induced by defibrillation shocks. AB - This study uses a one-dimensional model of cardiac strand to investigate the effects of electroporation on transmembrane potential (Vm) induced by defibrillation shocks. The strand is stimulated at the ends by extracellular electrodes. Its membrane, when exposed to large Vm, increases its conductance in a manner consistent with reversible electrical breakdown. Numerical simulations indicate that Vm increases proportionally to the shock strength only until the ends of the strand electroporate. Beyond this point, further increases in shock strength result in only a minor change in Vm. This arrest in the growth of Vm is caused by pores that develop in the cells immediately adjacent to the electrodes and that shunt part of the stimulating current directly into intracellular space. Consequently, only a fraction of the delivered current, Icr, gives rise to Vm; the current in excess of Icr divides itself proportionally between intra- and extracellular space and does not contribute to macroscopic Vm. Thus, electroporation has a beneficial effect: the formation of pores prevents the development of an excessively high Vm and limits the damage to the tissue. In contrast, electroporation does not affect the "sawtooth" component of Vm that reflects polarization of individual cells by electric field. These results indicate that electroporation does not impair the ability of the shock to reach the distant myocardium and may actually aid defibrillation by reducing nonuniformity of electrical conditions between regions close to the electrodes and in the bulk of tissue. PMID- 7491309 TI - Defibrillation thresholds are lower with smaller storage capacitors. AB - Present implantable cardioverter defibrillators use a wide range of capacitance values for the storage capacitor. However, the optimal capacitance value is unknown. We hypothesized that a smaller capacitor, by delivering its charge in a time closer to the heart chronaxie, should lower the defibrillation threshold (DFT). We compared the energy required to defibrillate 10 open-chest dogs, after 15 seconds of ventricular fibrillation, with a monophasic, time-truncated waveform delivered from either a 85-microF or a 140-microF capacitor. Shocks were delivered through a pair of 14-cm2 epicardial patch electrodes: The two capacitors were randomly tested twice with each dog using a modified 3-reversal method for each DFT determination. The average stored and delivered DFT energies for the 85-microF capacitor were 6.0 +/- 1.7 joules and 5.2 +/- 1.5 joules, respectively, compared to 6.7 +/- 1.7 joules and 6.0 +/- 1.5 joules for the 140 microF capacitor (P = 0.01 and P = 0.004, respectively). The mean leading edge voltages were higher, the pulse duration shorter, and the mean impedance lower for the 85-microF capacitor. The impedance was inversely related to the pulse duration and the voltage decay suggesting that, at least in part, the mechanism of improved defibrillation could be accounted for by the waveform electrical characteristics. There was an equal number of episodes of postshock bradyarrhythmias and tachyarrhythmias following discharges from each capacitor. Moreover, there was no relationship between the likelihood of these arrhythmias and either the initial voltage or the delivered current nor there was a higher number of episodes of postshock hypotension following the smaller capacitor discharges.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7491310 TI - Computer analysis of monophasic action potentials: manual validation and clinically pertinent applications. AB - Monophasic action potential (MAP) recordings are increasingly being used in a variety of clinical and experimental situations but their manual measurement is cumbersome, especially when hundreds or thousands of beats must be analyzed to monitor the exact time course of action potential duration (APD) changes following heart rate alterations, during surveillance of APD alternans, or during the onset and stabilization of Class III drug effects. To facilitate this task we developed a computer program that automates programmed electrical stimulation, digitizes at 1-kHz sampling frequency MAP recordings up to 8 channels simultaneously, analyzes all APDs at repolarization levels from 10%-90% in 10% decrements (APD10-90), and automatically outputs the analyzed numerical data into spreadsheets for graphical display or statistical analysis. To validate the computer algorithm, two independent observers manually analyzed 585 concurrent MAP recordings at a paper speed of 100 mm/s. Cycle length measurements by the computer were precise to 0.4 +/- 0.5 ms as compared to the computer determined paced cycle length. Computer measurements of APD20, 50, and 90 differed from manual measurements by 2.0 +/- 8.8 ms, 0.7 +/- 7.9 ms, and 0.2 +/- 8.5 ms, respectively, for observer 1; and by 12.2 +/- 8.3 ms, 5.8 +/- 7.5 ms, and 1.4 +/- 10.1 ms, respectively, for observer 2. Inter-observer variability (IOV) was 10.3 +/- 11.1 (APD20), 5.1 +/- 9.0 ms (APD50), and 1.2 +/- 7.8 ms (APD90), which was similar to computer/observer-2 differences and significantly greater (0.001) than computer/observer-1 differences. This indicates that the computer analysis was at least as precise as manual measurements when compared to IOV, and more precise when comparing computer/observer-1 differences to IOV. While providing equal or greater precision, computer-aided analysis of 100 MAP signals took approximately 1 minute while manual analysis of the same data set took between 2.5 and 4 hours. The pacing and analysis software was subsequently applied to experiments that mimic clinically pertinent examples of MAP recordings: (1) automatic generation, analysis, and graphical display of electrical restitution curves at multiple ventricular sites simultaneously; (2) evaluation of myocardial pharmacokinetics by monitoring the progression of Class III antiarrhythmic drug effects by continuous MAP recordings, and displaying differences in drug action between multiple sites; (3) depiction of the adaptation time course of APD to abrupt changes in paced cycle length; and (4) quantitative analysis of APD alternans during myocardial ischemia. The results show that our computerized algorithm greatly facilitates the generation of cardiac electrophysiological, and clinically important, data. PMID- 7491311 TI - Atrial pressure and experimental atrial fibrillation. AB - A possible profibrillatory effect on the atria of an elevated atrial pressure and the site of atrial stimulation was examined. In 15 anesthetized dogs, right or left atrial or biatrial pacing was applied at a high rate (300-600/min) for 5 seconds at double threshold intensity under a wide range of atrial pressures achieved by venous or arterial transfusion or bleeding. Induction of atrial fibrillation in 236 of 1,971 pacing runs was associated with a significantly higher (P < 0.001) atrial pressure (21.6 +/- 12.2 mmHg, mean +/- SD) than maintenance of sinus rhythm (16.8 +/- 11.1 mmHg in 1,735 of 1,971 pacing runs). Stimulation of the right atrium resulted in atrial fibrillation more frequently than left atrial or biatrial stimulation, with biatrial stimulation less frequent than right or left atrial stimulation. The induction of atrial fibrillation was related to the atrial pressure and to the site of stimulation but not to the pacing rate or the prepacing heart rate. The prepacing heart rate, associated with failure to induce sustained atrial fibrillation, was higher than that associated with atrial fibrillation in 12 of 15 experiments (significantly in 6) and not significantly lower in 3 of 15. Atrial fibrillation lasting 1 minute or more was more frequently associated with simultaneous stimulation of both atria than of either atrium alone. Thus, an elevated atrial pressure may facilitate the induction of atrial fibrillation. The site of stimulation also plays an important role for both the induction and maintenance of atrial fibrillation in this model. PMID- 7491312 TI - Basic concepts in cellular cardiac electrophysiology: Part II: Block of ion channels by antiarrhythmic drugs. AB - Antiarrhythmic drugs have relative specificity for blocking each of the major classes of ion channels that control the action potential. The kinetics of block is determined by the state of the channel. Those channel states occupied at depolarized potentials generally have greater affinity for the blocking drugs. The kinetics of the drug-channel interaction is important in determining the blocking profile observed clinically. The increased mortality resulting from drug treatment in CAST and several atrial fibrillation trials has resulted in a shift in antiarrhythmic drug development from the Na+ channel blocking (Class I) drugs to the K+ channel blocking (Class III) drugs. While both Classes of drugs have a proarrhythmic potential, this may be less for the Class III agents. Their lack of negative inotropy also make them more attractive. It is important that the potential advantages of these agents be evaluated in controlled clinical trials. In several laboratories, the techniques of molecular biology and biophysics are being combined to determine the block site of available drugs. This information will aid in the future development of agents with greater specificity, and hopefully greater efficacy and safety than those currently in clinical use. PMID- 7491313 TI - Software for electronic publishing. PMID- 7491314 TI - Redefining "experimental": a way out of the Medicare coverage dilemma. PMID- 7491315 TI - Missing atrial stimulus during DDD pacing. PMID- 7491317 TI - Preexcitation in hypertrophic cardiomyopathy: a case of a fasciculoventricular Mahaim fiber. AB - A patient with hypertrophic cardiomyopathy (HCM) who presented with preexcitation pattern on the surface ECG suggestive of the Wolff-Parkinson-White (WPW) syndrome is described. Intracardiac electrophysiological study revealed a fixed anomalous QRS complex and a short-fixed His-ventricular interval indicating a fasciculoventricular Mahaim fiber. As this specific form of accessory connection does not cause reentrant tachycardias, no treatment was required. It is important to distinguish this entity from atriofascicular or nodoventricular Mahaim fibers or the WPW syndrome in patients with HCM showing a preexcitation pattern in the surface ECG, as these may cause life-threatening arrhythmias in this patient population. PMID- 7491316 TI - Successful radiofrequency catheter ablation for macroreentrant ventricular tachycardias in a patient with tetralogy of Fallot after corrective surgery. AB - Radiofrequency (RF) catheter ablation was applied to two macroreentrant ventricular tachycardias (VTs) documented after corrective operation for tetralogy of Fallot. The activation wavefront of VT with a right bundle branch block pattern was found to revolve in a clockwise manner around a presumed myotomy scar in the right ventricle, and VT with a left bundle branch block pattern revolved around the same anatomical obstacle in a counterclockwise manner. In both VTs, the biggest conduction delay was confirmed at the right ventricular outflow tract. RF applications to the slow conduction area terminated each VT within a few seconds but were insufficient to cure the VTs. RF lesions were then applied to the slow conduction area in a line to intersect the macroreentrant circuit, and both VTs became noninducible. PMID- 7491318 TI - Cardiac involvement in Emery-Dreifuss muscular dystrophy: role of a diagnostic pacemaker. AB - Emery-Dreifuss muscular dystrophy is an X-linked recessive myopathy. Its progression is slow, and it rarely leads to cessation of walking; therefore, it has often been called "benign." On the other hand, cardiac involvement is often severe and sudden death is not uncommon. We describe a family with four affected males, two of whom died suddenly. The case of an affected man with first-degree AV block, incomplete right bundle branch block, and left anterior fascicular block is described in detail. The prophylactic insertion of a diagnostic pacemaker enabled us to follow the progress of conduction disturbances without leaving the patient unprotected. While AV and intraventricular conduction defects were more prominent before pacemaker implantation, sinus node dysfunction became more important during the follow-up. PMID- 7491320 TI - [Neonatal infections caused by group B beta-hemolytic Streptococcus]. AB - The aim of the Asiago Congress is to illustrate the progress in Group B Streptococcal neonatal disease management. It is of primary importance the obstetricians and neonatologists should think alike and should not allow their interest to develop along separate lines. The themes of the Congress were the incidence, the clinical and diagnostic new features, the old and the new therapeutic trends and the obstetrician's prevention. Contributions from twenty four Neonatologist Groups are included; they are summarized in four main articles and all together they form a synopsis of modern clinical practice and recent research in neonatal medicine. PMID- 7491319 TI - STIMAREC report. PMID- 7491321 TI - [Prevalence of group B beta-hemolytic Streptococcus colonization in a sample of 23,312 pregnant women and newborn infants]. AB - We report the prevalence of colonization of Group B Streptococci in a given population referred to a limited area in the north-west of Italy. 23.312 pregnant women were tested. Group B Streptococci have been isolated from genital cultures in 0.18-13.2% (mean 8.18). The prevalence of Group B streptococcal colonization from ear, throat and ocular cultures of newborn infants from colonized mothers was 11.55%. Incidence of infection in neonates has varied from 0 to 2.33% (1.5 per 1000 live births). PMID- 7491322 TI - [Clinical aspects and diagnosis of neonatal infections caused by group B beta hemolytic Streptococcus]. AB - Early-onset infection findings caused by Group B Streptococcus occur within 24 hours of birth (60 per cent of cases) but they may appear anytime during the first 5 days of life. In our experience early-onset infection affects both preterm and term neonates. The Authors report the usual clinical signs described for bacterial infections. Unusual findings are also reported: among 34 infants with early-onset infection, the congenital diaphragmatic hernia was associated with GBS septicemia in two neonates; beads of perspiration were the first only clinical finding in one neonate too. Two cases of late-onset infection are also reported. PMID- 7491323 TI - [Therapy of neonatal infection caused by group B beta-hemolytic Streptococcus]. AB - Expectant therapy for early Group B Streptococcus onset septicemia must provide coverage against other microorganism, such as L. Monocytogenes, H. Influenzae and S. Pneumoniae. It is possible to administer a combination of antimicrobial agents with activity against all or the most likely pathogens. Thus initial expectant therapy includes a broad spectrum semisynthetic penicillin (e.g. ampicillin) and an aminoglycoside (e.g. netilmicin). Vancomicin, teicoplanin and cefotaxime may also be used. Supportive therapy consists on temperature control, i.v. administration of fluids, acid-base balance and electrolytes monitoring, seizures control and ventilation. IV immunoglobulins, granulocyte and serum transfusion are also used. The G-Colony Stimulating Factor (G-CSF, filgastrim) usage is also reported. PMID- 7491324 TI - [Obstetric prevention of neonatal infections caused by group B beta-hemolytic Streptococcus]. AB - While our understanding of Group B Streptococcal infections has progressed with impressive measure, their prevention has not been accompanied by an effective means. Chemoprophylaxis for selected colonized women at rupture of membranes or at onset of labour, enhances benefit and minimizes adverse effects. Two or more maternal risk factors are special circumstances for routine use of chemoprophylaxis in asymptomatic neonates. Immunoprophylaxis by IgG antibodies directed against the type-specific polysaccharide antigen of GBS may be provided by passive or active immunization. Hyperimmune i.v. globulins or vaccination of adult women with low levels of antibodies in their sera have been demonstrate to be protective in vivo. PMID- 7491325 TI - [Genomic imprinting and human pathology. I. General Part]. AB - Genetical as well as experimental embryology methods have permitted to uncover a very important feature of mammalian embryonic development: it has been shown that female and male genomic complements are differentially imprinted in such a way that contribution of both a maternally and a paternally derived genome are absolutely necessary for the embryo to complete its normal development. The paternal and maternal genomes are not equivalent and have a complementary role during development in mammals. The differences in activity of each parental genome result from an epigenetic modification of the genome during gametogenesis: the parental imprinting. The recent discovery of several mouse and human genes which are imprinted should permit to address new data of some diseases. PMID- 7491326 TI - [Clinical-therapeutic aspects and surgical treatment of aortic coarctation associated with hypoplasia of the aortic arch in less than 3-month-old patients]. AB - The surgical treatment of the CoAo plus Hypoplastic Aortic Arch is debated. The point is whether to treat or not the associated tubular hypoplasia performing an extended Resection and "End to End" Anastomosis (ERETE) or a subclavian flap angioplasty (Waldhausen). According to some report in the literature we called an arch hypoplastic if its diameter in the first or second portion was less than the patient body weight plus 1. Between January 1988 and December 1992 in our Institution 38 patients less than 3 months underwent aortic coarctation repair. In 14 cases we were able to consider the aortic arch as hypoplastic. These patients were aged between 3 days and 45 days (median 15 days), the body weight was between 2.6 and 3.9 kgs (median 3.2 kgs). 6 patients had CoAo as isolated lesion, in 3 cases there was Ventricular Septal Defect (VSD) associated. In 5 patients major intracardiac anomalies were associated (1 DORV, 1 TGA, 2 SV, 1 SV+Arterio-Ventricular discordance). In 7 cases in order to treat the arch an ERETE was performed. The other 7 patients had a Waldhausen procedure. In 5 cases a Pulmonary Artery Banding (PAB) was associated, in 1 case a palliative arterial switch operation. The hospital death was 43% (3 patients) in the ERETE group, and 28% (2 patients) in the Waldhausen group. The echocardiography and or angiography performed during the follow-up period showed a nice growth of the arch in both groups. Although from our present data it's impossible to get definitive conclusions, the ERETE doesn't give better result in terms of hospital death and long term aortic arch growth.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7491327 TI - [Hepatitis B virus (HBV) infection in health personnel of a hospital for adolescents. Results of a vaccine campaign]. AB - The aim of the research is to evaluate the professional risk of the health workers of a pediatric hospital, mainly devoted to the care of adolescents. A seroepidemiological survey was therefore performed during the preliminary screening of an anti-hepatitis-B vaccination program, implemented in a suburban 150-beds pediatric hospital, with a predominance of elective surgery patients. The prevalence of past or ongoing HBV infection in a sample of adolescent inpatient (10-19 years old) in the same institution was known from an another study (13.2%). The prevalence of any HBV marker among 360 workers was 16.3%, lower in males (13.5 versus 17.7% in females), and in those with an higher level of instruction; the prevalence resulted significantly higher with increasing age and length of hospital service. Out of 321 candidates to anti-HBV vaccination 245 employees accepted the procedure (76.3%), significantly more females than males (80% versus 68.9%) and more subjects less than 30 years-old (84.3% versus 62% of subjects more than 40 years-old subjects). Only 13 subjects (5.3%) did not reach a protective titer after the third dose of the plasma-derived vaccine a protective titer. No major adverse event occurred. PMID- 7491328 TI - [Neuropsychological development of children born to HIV-positive mothers]. AB - 138 children born to HIV infected mothers (58 HIV infected and 80 seroreverted uninfected) were followed up at the Institute of Infectious Diseases of the University of Pavia. All patients had an evaluation of neurological assessment every three months done by a pediatric neurologist (Amiel-Tison and Towen) and an evaluation of psychological assessment by observation of child-mother interaction. The development quotient was examined using the Griffiths Scales and mental development using the Wisc-R scale. The incidence of Central Nervous System involvement was 36% in children with symptomatic HIV infection, the hump of incidence of severe encephalopathy was in the first 4 or 5 year of life, another hump (smaller) was in preadolescence. Development deficit and psychological problems were discovered both in asymptomatic infected and in non infected children, without difference of percentage between the two groups. PMID- 7491329 TI - [Continuous intraesophageal pH monitoring in the study of gastroesophageal reflux (GER) in children. Influence of age on the considered parameters]. AB - Gastroesophageal reflux is a very common event in children; it is studied using 24-hour intraesophageal pH-monitoring, which has a large sensitivity and specificity, but in the literature is not clear if the age of the children could have influence on the parameters considered in the pH-monitoring. The aim of our study is to assess if the age of our small patients could modify the results of pH-monitoring. During 3-year period from 1991-1993 continuous intraesophageal pH monitoring of 45 children (29 males and 16 females), divided in three groups on the bases of age, homogeneous for number, sex and clinical symptoms, with pathologic reflux time (> 4% of 24 hours) has been evaluated. In our cases the pathologic total reflux time in 24 hours doesn't seem change with age. It has been noted that under two years of age the pH-monitoring measures are homogeneous with a tendency of reflux longer than children over two years. This older group has more after lunch refluxes; that is the number of total refluxes is larger in older children but the number of refluxes with length higher than five minutes is more in younger. These data are discussed in relation to the anatomical and functional gastroesophageal characteristics of the different age of the child. PMID- 7491330 TI - [Evaluation of 3-year ultrasound screening in congenital hip dysplasia. Comparison of clinical examination, risk factors and ultrasonography. Interpretation of Graf type IIA grading and the most suitable age for 1st ultrasonography]. AB - 2237 infants (1210 females, 1027 males) aged two days-six months were screened by ultrasound for congenital dysplasia of the hip (CDH). 21 infants (0.93%) were detected having CDH by ultrasound examination. We have evaluated the concordance between ultrasonography, clinical examination (Ortolani's examination) and risk factors. Of the 21 infants with CDH, 10 (47.6%) had a positivity of Ortolani's examination, 13 (61.9%) had at least one risk factor, 3 (14.2%) had a negative clinical evaluation and no risk factors. Our data indicates the superiority of ultrasonography as compared with physical examination alone, for the diagnosis of CDH. Infants presenting any risk factors for CDH and/or a positive Ortolani's examination should have an early ultrasound evaluation in order to allow initiation of treatment early on, while in infants with negative clinical findings and no risk factors, it should be postponed to avoid the frequent finding of physiological immaturity of the hip. The finding of a high rate of infants that presented a grade IIA in the Graf's grading system during their first month of life and that evolved favourably in the absence of risk factors, and which required further sonograms, indicates the period between the second and the third month as the most desirable for ultrasound screening of CDH. PMID- 7491331 TI - [Metabolic, hormonal and nutritional effects of long-term theophylline therapy in preterm infants: a case-control study]. AB - Theophylline is widely used in preterm newborns for the prevention of idiopathic apnoeas, but few controlled studies have evaluated its effects on the nutritional and hormonal status of the infant. For this reason we have studied the effect of long term theophylline administration on 16 laboratory parameters concerning the metabolism of proteins, glucose, lipids, hormones and the glomerular function (blood: hemoglobin, glucose, albumin, prealbumin, urea nitrogen, creatinine, cholesterol, triglycerides, apolipoproteins A-I and B-100, IGF-I, IGFBP-3; urine: urea nitrogen, creatinine, C-peptide, GH). A case-control study was performed on 18 healthy preterm infants who were receiving oral theophylline for the prevention of idiopathic apnoeas. The mean duration of therapy at the moment of the balance study was 31 days (SD 12, range 12-51), the mean daily dose was 4.2 mg/kg (SD 1.0), the plasma range of theophylline concentration was 5 to 15 mg/l. As controls, 18 healthy preterm infants of comparable post-conceptional age, body weight and calories/protein intake at the moment of the study, were selected if they had been never treated with theophylline. No statistically significant differences were found between the two groups for the growth velocity or any of the parameters studied. The only notable exception was hemoglobin, which was significantly lower in theophylline treated infants (mean values 10.5 vs 12.7 g/dl, p 0.005 at t test). In synthesis, long term theophylline treatment in preterm infants seems to be safe from the point of view of growth, glucose, protein and lipid metabolism, hormones and glomerular function, but further studies are needed on the effects of theophylline on neonatal erythropoiesis. PMID- 7491332 TI - [Dietetic counseling of obese school child. Study in an elementary school]. AB - We controlled, after nine months, 29 obese children, who attended the primary school, and to whom it was advised a good balanced base diet of 1200 kcal. One child wasn't obese anymore, and in 13 cases there was a sensible reduction of overweight percentage. We consider these results positively, because our aim was to obtain a gradual resolution of the overweight, trying to avoid some very dangerous relapses for the young's future. Finally, we underline the necessity to begin some conferences about the young's and family's alimentary education, as soon as possible. These conferences have to be addressed to all the teachers, parents and pupils, from the primary school. PMID- 7491333 TI - [Sotos syndrome: follow-up of a case with precocious puberty]. AB - Sotos syndrome is a rare disorder of statural overgrowth in children. Peculiar facies, pre- and post-natal overgrowth and advanced bone age are the main clinical findings; developmental delay may be present. The Authors report a case of Sotos syndrome followed until adult height and describe the pattern of growth. The girl showed the typical findings of the syndrome together with premature pubarche and premature pubertal development. Since precocious puberty determines an increase of growth during childhood, the Authors suggest that Sotos syndrome should be considered and excluded in girls with premature pubertal development. PMID- 7491336 TI - [Pseudolithiasis caused by ceftriaxone in children: a case report]. AB - Recently, there are in literature some reports concerning the sonographic observation of transient gallbladder precipitates in children and adults during ceftriaxone treatment. The condition is defined as ceftriaxone-reversible "pseudolithiasis" and the gallbladder precipitates resolve spontaneously after cessation of therapy. We present a case of a 1 year old children treated with ceftriaxone who presented asymptomatic pseudolithiasis that resolved partially two weeks after cessation of treatment. The sonographic observation performed four months after cessation of therapy show a normally gallbladder, without precipitates or sludge. PMID- 7491335 TI - [A case of Hallermann-Streiff syndrome with rapidly fatal course]. AB - The Hallermann-Streiff syndrome is a rare affection characterized by beaked nose, dyscephaly, hypotrichosis, cataracts, micrognathia and proportionate short stature. The most severe complication in the syndrome is respiratory embarrassment. Narrow air passage with abnormal glottic closure, prenatal growth deficiency, immunodeficiency and sometimes associated cardiovascular anomalies, can predispose these patients to pulmonary infections. The Authors describe a new born with Hallermann-Streiff syndrome, who, after a brief period of obstructive apnea successfully treated with CPAP, developed a severe pulmonary infection that caused his death at the age of 61 days. PMID- 7491334 TI - [Acetazolamide treatment of a newborn infant with posthemorrhagic cerebral ventricular dilatation]. AB - We report a case of a male newborn delivered by vacuum at 40 weeks of gestation, who presented perinatal asphyxia, neonatal seizures and ventricular posthemorrhagic dilatation. The newborn was treated with Acetazolamide for 45 days until the reduction of the ventricular dilatation. This case shows the efficacy and the absence of collateral effects of Acetazolamide. This therapy suggests that it may be used as an alternative or an adjunct to lumbar punctures. The pharmacol treatment in the posthemorrhagic ventricular dilatation has not been well studied and there are a few related cases in the literature. PMID- 7491337 TI - [Skin lesions caused by oxygen saturation monitor probe: pathogenetic considerations concerning 2 neonatal cases]. AB - Two cases of skin injury by a pulse oximeter probe are reported. The Authors hypothesize the local pressure as pathogenetic mechanism of the skin lesion and emphasize the importance of a careful management of critically ill patients during non-invasive monitoring. PMID- 7491343 TI - [Cytokines (interleukin 1 and 6, tumor necrosis factor) and their possible role in infective endocarditis]. PMID- 7491341 TI - [Pertussis vaccination]. PMID- 7491338 TI - [Hemangioma of the bladder in children: 2 case reports and review of the literature]. AB - Hemangioma of the bladder is a rare benign tumor. Hematuria is the most common symptom of the disease. The Authors report their experience on two isolated cases of bladder hemangioma. The lesions were diagnosed by endoscopy and biopsy. In both cases the hemangiomas were electrocoagulated endoscopically. Follow-up examination after 1 year, has shown complete eradication of the lesion. PMID- 7491340 TI - [Gastroesophageal reflux (GER)]. PMID- 7491339 TI - [Cholelithiasis in children: observations on a further case and the technique of laparoscopic cholecystectomy in a child]. AB - A new case of gallstone in a child, with non apparent predisposing condition, who underwent laparoscopic cholecystectomy is reported. The importance of considering cholelithiasis in children with abdominal pain as well as the value of ultrasonography in diagnosis is stressed. The technique of laparoscopic cholecystectomy modified for pediatric patients and its benefits (decreased pain and ileus after surgery, shortened hospitalization and improved cosmesis) are discussed. PMID- 7491342 TI - [Opinion of the expert]. PMID- 7491344 TI - [The role of "hemochromatosis alleles" in the pathogenesis of polyglobulia after renal transplantation]. AB - 43 kidney recipients were followed up for 12 months after renal transplantation. In 13 patients treated with cyclosporin A for 6 months polyglobulia was found, which subsided after conversion of immunosuppressive therapy. A highly significant correlation was found between the presence of the antigen A3 and the polyglobulia in kidney recipients treated by cyclosporin A. The serum ferritin level was not elevated in kidney recipients who were carriers of the "hemochromatosis alleles". Results obtained in this study seem to justify the following conclusions: 1. Kidney recipients who are carriers of "hemochromatosis alleles" show a threefold higher risk of posttransplant polyglobulia. 2. Presence of "hemochromatosis alleles" is not an absolute predictor of polyglobulia in patients after kidney transplantation. 3. Both etiology and pathogenesis of posttransplant polyglobulia do not seem to be homogeneous. Cyclosporin A, in contrast to azathioprine, seems to have a promoting effect on the development of posttransplant polyglobulia. Therefore treatment of kidney transplant patients with cyclosporin A, who are carriers of "hemochromatosis alleles", needs careful monitoring. PMID- 7491345 TI - [In vitro studies for evaluating the influence of recombinant interleukin-11 on human erythropoiesis. Potential clinical implications]. AB - According to the experimental data that IL-11 costimulates the proliferation of human hematopoietic progenitors in vitro the first clinical trials in vivo have been performed. Surprisingly, the patients treated with recombinant Il-11 display decrease in the hematocrit values. This finding is contradictory to previous experimental studies in vitro, where IL-11 was found to be a potent costimulator of eytrhopoiesis. Therefore in order to obtain our own data the influence of IL 11 on human erythropoiesis in vitro has been reevaluated. In performed studies we did not observed any influence of IL-11 on the cloning efficiency of human erythropoietic progenitors in vitro. Our results demonstrate on one side that clinical use of IL-11 in order to stimulate erythropoiesis is not enough theoretically grounded and on the other that the decrease of hematocrit values observed after in vivo treatment with recombinant IL-11 could not be explained simply by the fact of the possibility of the direct inhibition of erythroid colonies formation by Il-11. PMID- 7491346 TI - ["Prognostic markers" in serum of active and passive intravenous drug users infected with human immunodeficiency virus (HIV)]. AB - The analyse of neopterin (NPT), beta 2-microglobulin (beta 2-M) and immunoglobulin A (IgA) in active and passive intravenous drug users was made to show the influence of intravenous drug taking on the level of so called "prognostic markers" in the serum of HIV infected persons. The intravenous drug users remained in the same, symptom-free stage of infection. The level of NPT in active drug users was higher than in passive drug users (statistically essential difference). In both analysed groups the level of NPT considerably exceeded the norm range (10 nmol/l). The differences in beta 2-M concentrations between analysed groups were not statistically essential, although higher values were observed in active drug users. The concentration of IgA in analysed persons were within the norm range. Current life conditions of the HIV-infected persons should be taken into consideration when estimating prognostic values of analysed indices. PMID- 7491348 TI - [High levels of cholesterol and lipoprotein (A) in serum decreases the inhibitory effect of aspirin on generation of thrombin]. AB - Aspirin (ASA) is widely used in the treatment of cardiovascular diseases. Recently, we have found that aspirin decreases not only platelet aggregation but also thrombin generation. This effect, however, was seen only in certain subjects. Therefore we decided to examine influence of a single dose of aspirin (500 mg) on thrombin generation in healthy volunteers. Thrombin genesis was assessed by serial measurements of fibrinopeptide A concentration in blood emerging from standardised forearm skin incisions. Aspirin reduced thrombin generation in persons with normal serum level of lipids. This effect was lost, however, in subjects with high level of cholesterol and lipoprotein (a)--well known risk factors of ischaemic heart disease. While the mechanism by which aspirin affects thrombin generation remains to be elucidated, our data indicate that hypercholesterolemic subjects might benefit less than others from preventive aspirin treatment. PMID- 7491347 TI - [The effect of indobufen on platelet function and dialyzer regeneration in patients on hemodialysis]. AB - In 16 patients on chronic haemodialysis treatment the platelet activation and function were studied during 2 weeks antiplatelet therapy with indobufen. Followings tests were made: platelet count, platelet aggregation, platelet factor 3 and 4. Indobufen inhibited platelet function, mainly release of platelet factor 4 and improved dialyser regeneration. PMID- 7491349 TI - [Predictors of survival in patients with chronic obstructive pulmonary disease with moderate hypoxemia]. AB - Long-term oxygen therapy (LTOT) is a well established method of treatment in patients with chronic obstructive pulmonary disease (COPD). There are two main qualification criteria for LTOT: severe hypoxaemia (PaO2 < 55 mmHg) and moderate hypoxemia (PaO2 > 55 mmHg) accompanied by signs of cor pulmonale. We were unable to find any controlled investigations on the effect of LTOT on survival in this group of patients. We studied 85 COPD patients with PaO2 between 56-65 mmHg. Fourty four patients were treated at theirs homes with LTOT, 41 formed a control group. Both groups presented with severe airway obstruction--FEV1 < 1L. All patients were followed up for at least 3 years. During this time 27 patients died from the LTOT group and 26 patients from the control group. Survivors were younger and had less severe airway obstruction than nonsurvivors. Cox's proportional hazard regression analysis showed that only age and spirometric parameters--VC, VC%N, FEV1 were good predictors of survival. LTOT did not improve prognosis in this group of patients. Results of our study suggest the need of verification of LTOT qualification criteria. PMID- 7491351 TI - [Pathogenic mechanisms and natural history of pulmonary hypertension in chronic obstructive pulmonary disease]. PMID- 7491352 TI - [Value of prophylactic use of lidocaine in acute myocardial infarction]. PMID- 7491350 TI - [Effects of home oxygen therapy on pulmonary hemodynamics in patients with chronic obstructive pulmonary disease]. AB - The aim of the study was to assess effects of long-term oxygen therapy (LTOT) on pulmonary haemodynamics in patients with chronic obstructive pulmonary disease (COPD). 93 pts (73M, 20F) mean age 58 years, entered the study. Their functional characteristics at entry for LTOT revealed: VC 2.33L, FEV1, 0.85L, FEV1/VC 37%, PaO2 55.5 mmHg, PaO2 46.7 mmHg, Hct 50%. Pulmonary haemodynamics showed: mild pulmonary hypertension (mean pulmonary arterial pressure PPA 25.9 mmHg), normal pulmonary wedge pressure (Pw) and increased driving pressure (PPA-Pw) 17.8 mmHg. Cardiac output (CO) was normal and pulmonary vascular resistance (PVR) averaged 313 dyne.s.cm-5. All enrolled patients were treated with oxygen at home and followed-up for at least 2 years. After 2 years of LTOT PPA and Pw fell by 2.4. and 2.6 mmHg, respectively, and (PPA-Pw) remained stable. An increase in CO by 0.31.min-1 was found while PVR was stable. After 4 years of treatment PPA, (PPA Pw) and CO increased by 1.5, 1.7 mmHg and 0.9 l.min-1 respectively, while Pw and PVR remained stable. In 12 six years survivors PPA decreased after 2 years by 4.4 mmHg and cardiac output increased by 2 l.min-1 both returning to baseline after 6 years of LTOT. No significant changes in PVR were observed. Comparison of lung function tests before and after 6 years of LTOT showed deterioration of airways obstruction and of respiratory failure. LTOT for 14 h/d results in long-term stabilization of the hypoxic pulmonary hypertension despite of the progression of the natural course of COPD. PMID- 7491353 TI - [Congenital methemoglobinemia found in an adult. Case report]. AB - Cyanosis in a 36 year old patient which persisted 20 years after a successful surgical closure of her patent foramen ovale has been finally diagnosed as due to congenital methemoglobinemia: a 28% level of methaemoglobin and no activity od NAD-dependent methaemoglobin reductase were found. Erythrocyte antioxidative system was studied i.e. glutathione peroxidase, reductase, transferase, superoxidase dismutase and glucose-6-phosphate dehydrogenase. Increased activity was disclosed of superoxide dismutase and glucose-6-phosphate dehydrogenase in erythrocytes in comparison to normal individuals as well as increased concentration of lipid peroxidation products coexisting with methaemoglobin reductase deficiency. PMID- 7491355 TI - [The cytoskeleton of muscle cells]. PMID- 7491354 TI - [Certain biochemical aspects of pathogenesis in stunned myocardium]. PMID- 7491356 TI - [MAP-2 protein and its role in several aspects of plasticity and pathologic states in the central nervous system]. PMID- 7491357 TI - [Inherited AMP-deaminase deficiency--molecular basis of the defect]. PMID- 7491358 TI - [Platelet collagen receptors]. PMID- 7491359 TI - [Interactions of proteins with damaged DNA]. PMID- 7491360 TI - [Biological consequences of lipoprotein modification]. PMID- 7491361 TI - [Preparation of highly-purified RNA from yeasts using calcium ions]. AB - The influence of temperature and Ca2+ ions at different concentrations on the effectiveness of RNA precipitation from water-salt extract of baker yeast was studied. Our findings allowed us to propose a method for producing highly purified RNA by precipitation with 180 mM Ca2+ ions from hot yeast extracts (95 degrees C) after removal of the protein fraction that could be also precipitated with 20 mM Ca2+ at 90 degrees C. The purity and yields of RNA samples obtained in such a manner was of 98% and 56% relative to theoretical values, respectively. PMID- 7491362 TI - [Alkaloids from Penicillium species fungi isolated from food products]. AB - Alkaloid- and toxin-forming ability of 18 strains of Penicillium fungi, isolated from foodstuffs, is studied. It is shown that all cultures are producers of alkaloids and mycotoxins that belong to different structural groups. The composition of isolated metabolites, synthesized under submerged and surface cultivation on complex and synthetic media, is established. PMID- 7491363 TI - [Changes in the lipid composition of salmon liver and eye lens in cataracts]. AB - The study of pathogenesis of cataract in salmon yearlings revealed changes in certain fractions of phospholipids and fatty acids in the eye lens and liver of specimens with and without cataract. The level of malonic dialdehyde in these tissues was increased, indicating an increased rate of lipid peroxidation. PMID- 7491365 TI - Noncompetitive NMDA receptor antagonists with fast open-channel blocking kinetics and strong voltage-dependency as potential therapeutic agents for Alzheimer's dementia. AB - There is a growing body of evidence that disturbances of glutamatergic neurotransmission may underlie the pathomechanism and cognitive deficits of Alzheimer's disease. This review describes the potential use of low affinity, noncompetitive NMDA receptor antagonists in the treatment of this disease using memantine as an example. Evidence is presented indicating that this class of compound is neuroprotective in preclinical models of subchronic glutamate toxicity without producing side effects characteristic for other classes of NMDA receptor antagonist. This is attributed to their fast blocking kinetics and strong voltage dependency. Memantine also produces symptomatological improvement of cognition in animal models. The mechanism of action of this effect is still unclear but might be related to an enhancement of AMPA receptor mediated neurotransmission. In patients with dementia syndrome of various aetiologies, memantine produces a rapid onset, clinical improvement in various symptomatological deficits. PMID- 7491364 TI - The Association of Neuropsychopharmacology and Pharmacopsychiatry (AGNP): proposed issues for a residency's training in psychopharmacology (Part 1). PMID- 7491366 TI - Confirmed clinical efficacy of Actovegin in elderly patients with organic brain syndrome. AB - A double-blind randomized clinical trial was performed comparing the therapeutic effects of Actovegin versus placebo in elderly patients with organic brain syndrome. In addition to the necessary basic internal medicine therapy, 40 geriatric patients received dialy intravenous infusions of 250 ml Actovegin 20% p.i., and 20 patients received 250 ml 0.9% saline solution as placebo over a period of four weeks. Of the patient sample, 58% were hospitalized for simple dementia (ICD-9: 290.0) and 42% due to senile dementia with depressive or paranoid symptoms (ICD-9: 290.2). Based on the Syndrome Short Test (SKT) and the Sandoz Clinical Assessment Geriatric Scale (SCAG) score, the patients suffered from mild to moderate dementia. The therapeutic effect on the total SCAG score and the Clinical Global Impression (CGI) were the primary study variables. The scores for the SCAG subscales and the SKT score served as secondary variables. The mean total SCAG score in the drug group decreased from 56.3 at the start of therapy to 36.3 points at the end of therapy, and in the placebo group the total score went from 61.2 to 52.0 (p < 0.01). The CGI showed that with Actovegin, 70% of the patients experienced "distinct improvement" or "improvement" compared to only 35% with such results in the placebo group. The SCAG subscales and the total SKT score also demonstrated the superior effects of Actovegin compared to placebo. Moreover, the therapy group treated with Actovegin showed greater improvements in social behavior and mental performance than did the placebo group.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7491368 TI - Reduced beta-amyloid response in lymphocytes of patients with Alzheimer's disease. AB - The beta-amyloid peptide (beta A4) is a principal constituent of senile plaques and is thought to play a major role in the pathophysiology of Alzheimer's disease (AD). Although the mechanism of beta-A4 neurotoxicity is still a matter of debate, one of its effects might be a destabilization of cellular calcium homeostasis, thus promoting neuronal damage. The influence of the toxic fragment beta A25-35 on the mitogen-induced rise in the intracellular calcium concentration ([Ca2+]i) in lymphocytes of AD (n = 13) and depressive patients (n = 14) as well as in healthy controls was therefore investigated (n = 16). The results showed a significant increase in the mitogen-induced calcium signal with lymphocytes of healthy controls and depressive patients. This beta A25-35-induced amplification was significantly lower in AD patients as compared to healthy controls but not as compared to depressive patients. The results thus confirm a postulated decreased beta-amyloid sensitivity in AD lymphocytes. However, this effect might not be as pronounced or as specific as recently described by Eckert et al., (1993b). PMID- 7491367 TI - Cognitive psychophysiology in nootropic drug research: effects of Ginkgo biloba on event-related potentials (P300) in age-associated memory impairment. AB - Extracts from the leaves of Ginkgo biloba have been suggested to be useful in the treatment of various symptoms of impaired brain functions in advanced age. To elucidate specific mechanisms of the possible clinical benefit, the effects of Ginkgo biloba extract Ginkobene on cognitive information-processing were investigated by means of long-latency auditory event-related potentials. In a double-blind placebo-controlled study, 48 patients (29 women and 19 men) aged between 51 and 79 years with the diagnosis of age-associated memory impairment had 57 days' treatment with a daily dosage of 3 x 40 mg Ginkobene or placebo. To evaluate acute, chronic, and superimposed drug effects, psychophysiological investigations were carried out on day 1 and day 57 before and 3 hours after drug administration. ERP investigations were carried out by means of a two-tone auditory oddball paradigm. In addition to 17 EEG leads, vertical and horizontal EOGs were recorded. After minimizing ocular artifacts and visual artifact rejection, latencies and topographic distributions of N1 and P2 components (non targets) and N2 and P300 components (targets) were calculated by an automatic procedure. When compared to the placebo group, in the Ginkobene group no consistent and unequivocal changes on N1, P2, N2, and P300 amplitudes or on N1, P2, and N2 latencies were observed. P300 latency was shortened by 31 ms, 38 ms, and 32 ms in the Ginkgo biloba group after acute, chronic, and superimposed drug administration. It may therefore be hypothesized that the decrease of P300 latency in the Ginkgo biloba group may reflect shorter stimulus-evaluation time.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7491369 TI - Treatment of chronic depression with sertraline or imipramine: preliminary blinded response rates and high rates of undertreatment in the community. AB - Despite the prevalence of chronic depression and its associated morbidity, there has been little systematic study of pharmacotherapy for this disorder. In this article, we report a preliminary analysis of the first 12-week phase of a multicenter clinical trial that will eventually include approximately 635 patients in acute, continuation, crossover, and maintenance studies of sertraline, a selective serotonin reuptake inhibitor (SSRI), and imipramine, a tricyclic antidepressant, for the treatment of chronic depression. Of the first 212 patients to enter the study, 168 completed all 12 weeks; of these, 61.3 percent were responders, including 58.9 percent of the 73 patients with chronic major depression and 63.2 percent of the 95 patients with double depression. Only 26.8 percent of the 198 patients for whom such data were available had ever had an adequate trial of an antidepressant medication, defined as 150 mg/day of imipramine or its equivalent taken for at least 4 consecutive weeks. In general, demographic and diagnostic characteristics were more similar than different for patients with chronic major and double depression. However, comorbid generalized anxiety disorder was significantly more common in patients with chronic major depression (11.2% threshold for chronic versus 4.9% threshold for double depression, p = .02). The results of this study provide preliminary evidence of the responsiveness of patients with chronic major or double depression to an SSRI or a tricyclic antidepressant. PMID- 7491370 TI - Stability of remission during tricyclic antidepressant continuation therapy for dysthymia. AB - Sixty-four of 73 patients with DSM-III-R dysthymia who responded to short-term treatment with tricyclic antidepressants (TCAs) completed 16 to 20 weeks of continuation therapy on the same medication. Forty-one of 46 full remitters remained in full remission, while 5 became partial remitters during the continuation treatment. Of 18 partial remitters, 8 became fully remitted, 8 remained partial remitters, and 2 relapsed. Thus the stability of remission in dysthymic subjects treated with TCA seems quite favorable during continuation treatment. PMID- 7491371 TI - Comparative neurotransmitter reuptake and anticholinergic potencies of the 8 hydroxy metabolites of clomipramine. AB - We hypothesize that 8-hydroxy-clomipramine (8-OH-CMI), the major hydroxy metabolite of clomipramine (CMI), may have antidepressant properties with less anticholinergic potency than CMI. We compared the potencies of 8-OH-CMI and CMI for inhibition of serotonin and norepinephrine reuptake and the potencies of these compounds for blockade of muscarinic receptors. We also compared the antimuscarinic potencies of desmethylclomipramine (DCMI) and 8-hydroxy desmethylclomipramine (8-OH-DCMI). We found that 8-OH-CMI inhibits the uptake of serotonin and norepinephrine to the same extent as CMI and that 8-OH-CMI has far less antimuscarinic potency than CMI. We also found that 8-OH-DCMI has about one tenth the antimuscarinic potency of DCMI. Since the therapeutic efficacy of CMI may be related to its effect on the reuptake of neurotransmitters, and since the extent of clinical anticholinergic effects of tricyclic antidepressants has been shown to be related to their in vitro antimuscarinic potencies, these results raise the possibility that 8-OH-CMI may be an analogue of CMI with fewer anticholinergic side effects than the parent compound. PMID- 7491372 TI - Patterns of improvement in depressive symptoms with fluoxetine treatment. AB - By definition, patients who respond to antidepressant treatment show significant improvements in most, if not all, of their depressive symptoms. We assessed the patterns of change in depressed outpatients treated openly with fluoxetine 20 mg/day for 8 weeks. We studied 62 depressed outpatients with major depressive disorder who showed full response following fluoxetine treatment. Analysis of the data revealed that the symptoms that tended to improve relatively early during treatment were suicidal ideation, excessive guilt, and lack of appetite. Our tentative conclusion from this pilot study was that symptom improvement tends to occur in both cognitive and neurovegetative areas early in treatment as well as later. PMID- 7491374 TI - Onset of menses in two adult patients with Prader-Willi syndrome treated with fluoxetine. AB - Prader-Willi syndrome (PWS) is characterized by hypotonia at birth, hypogonadism, early childhood obesity, and mental deficiency. Hypogonadotropic hypogonadism is a major characteristic of patients with PWS, and it is speculated to be due to hypothalamic insufficiency. Two adult female patients with PWS and no prior history of menses are presented. Both of these patients were treated with fluoxetine for psychopharmacologic management of obsessive features in the form of food preoccupation and hyperphagia or for compulsive behaviors in the form of severe self-injurious behaviors. The two female patients with PWS who had primary amenorrhea developed vaginal bleeding believed to be menses following at least 6 months of treatment with fluoxetine. Mature hypothalamic function is characterized by pulsatile release of gonadotropin-releasing hormone (GnRH) in a critical range of frequency and amplitude. Central nervous system neurotransmitters may modify GnRH secretion. Fluoxetine specifically inhibits the reuptake of serotonin which may impact the hypothalamic-pituitary-ovarian system in female patients with PWS. PMID- 7491373 TI - The effects of fluoxetine on symptoms of insomnia in depressed patients. AB - Fluoxetine is an efficacious, nonsedative antidepressant, but its selective efficacy on symptoms of insomnia has not been thoroughly explored. In this analysis, the effects of fluoxetine versus placebo on symptoms of insomnia were evaluated in three clinical subgroups: patients with baseline sleep disturbance, melancholia, or reduced rapid eye movement (REM) latency. Eighty-nine outpatients with major depression completed 2 nights of polysomnography (PSG) and were randomized to fluoxetine or placebo. Within each subgroup of patients, fluoxetine was statistically significantly more effective than placebo in improving non sleep Hamilton Rating Scale for Depression (HAM-D) items (HAM-D-17 total minus scores from Items 4, 5, and 6). Numerical improvement in HAM-D sleep total (sum of HAM-D Items 4, 5, and 6) was also seen for fluoxetine versus placebo. Fluoxetine did not exacerbate sleep disturbance either at Week 1 or at endpoint. These findings suggest that fluoxetine is an effective antidepressant in patients with baseline sleep disturbance, melancholia, and reduced REM latency. PMID- 7491376 TI - The use of decision analysis in the pharmacoeconomic evaluation of an antidepressant: a cost-effectiveness study of nefazodone. AB - A decision model can be a useful tool for evaluating the cost-effectiveness of a drug during research and development, before the drug is marketed. Decision analysis provides a structured process for comparing the costs and consequences of a new drug with those of standard drug therapy. This article introduces the use of a decision model and discusses the advantages and disadvantages of such a model. A cost-utility study of nefazodone, a new antidepressant, illustrates the application and interpretation of decision models. Decision analyses using data from clinical trials are a great potential source of information on the economic impact of new drugs. However, the development and use of such models require tolerance of uncertainty, the ability to represent complex relationships accurately, awareness of all factors that might influence the results, and a willingness to validate the model to the extent that validation is possible. The disadvantage of clinical decision models is that the findings are only as reliable as the information on which the model is based. Nevertheless, sensitivity analyses can be used to vary any information in question and assess the impact of that variation on the findings of the analyses. The advantage of clinical decision models is that they encourage the consideration and explicit representation of all pertinent inputs and outcomes. They clearly differentiate knowledge supported by data from assumptions, and compel assessment of the effect of those assumptions on the findings. The cost-effectiveness of nefazodone relative to standard treatment for major depression was evaluated using a decision model that simulated the lifetime direct medical costs and health outcomes associated with nefazodone and two comparators, imipramine and fluoxetine. The model included the possible clinical management pathways and the use of medical resources in treating major depression from early adulthood until death. Sensitivity analyses were performed to determine the impact of variation in model parameters on the results of the model. Using the base case assumptions of the model, which included parity of pricing with fluoxetine, nefazodone was found to be a cost-effective treatment for major depression. PMID- 7491377 TI - Antidepressant-induced tardive dyskinesia: review and case report. AB - We report a case of clomipramine-induced tardive dyskinesia (TD) in the setting of chronic use of dextroamphetamine without prior use of neuroleptics, in which the movements persisted after discontinuation of the clomipramine. Other contributing factors include advanced age, history of alcohol abuse, and concomitant administration of hepatic enzyme inhibitors. Other cases of tricyclic induced TD have occurred primarily in combination with neuroleptics, have involved antidepressants with antidopaminergic actions, or, as in the present case, have resulted from antidepressants with significant anticholinergic effects (n = 17). Predisposing risk factors and potential mechanisms in the precipitation of dyskinesias are discussed. PMID- 7491375 TI - Amplitude/intensity functions of auditory event-related potentials predict responsiveness to bupropion in major depressive disorder. AB - Patients diagnosed with major depressive disorder (MDD) and enrolled in an open label safety surveillance study of a sustained release formulation of bupropion hydrochloride (100 to 300 mg/day) were evaluated with the Hamilton Rating Scale for Depression (HAM-D) immediately before and 6 to 12 weeks after the initiation of drug treatment. Auditory event-related potentials (ERPs) recorded under a stimulus intensity modulation paradigm were also obtained at these times. Patients were classified as responders and nonresponders based on post-treatment HAM-D scores, with responders having HAM-D scores less than 10 and nonresponders having scores greater than 10. Consistent with our previous findings, responders exhibited significantly larger positive slope coefficients for P2 ERP component amplitudes as a function of auditory stimulus intensity obtained at baseline and were not affected by bupropion treatment. Thus, these results further support our previous finding that ERP amplitude/intensity functions measured under a stimulus intensity modulation paradigm provide information about the likelihood of a positive therapeutic response to antidepressant pharmacotherapy in patients with MDD and extends these results to bupropion, a pharmacologically atypical antidepressant agent. PMID- 7491378 TI - Lithium combined with carbamazepine or haloperidol in the treatment of mania. AB - Hospitalized manic patients were withdrawn from psychoactive medications for 2 weeks after which they were randomized to double-blind treatment with carbamazepine plus lithium [CBZ-Li] or haloperidol plus lithium [HAL-Li] with benztropine. Unit dosages of Li 300 mg, CBZ 200 mg and HAL 2 mg were titrated to therapeutic plasma levels and maintained for 8 weeks. No rescue medications were permitted after 3 weeks. Standard ratings of psychopathology and side effects were accomplished weekly. Sixty patients entered the study but only 33 remained for randomization after drug washout. By 8 weeks both groups were improved from baseline without statistically reliable differences between them. However HAL-Li patients had more extrapyramidal side effects that were major reasons for dropout, whereas CBZ-Li patients were more often noncompliant and initially required more rescue medications. We conclude that either combination treatment can be beneficial but CBZ-Li has the advantage because of fewer neurologic side effects. PMID- 7491379 TI - Pattern of placebo response in panic disorder. AB - This study characterizes the pattern of placebo response in depressed panic disorder patients who participated in a randomized clinical trial. We compared placebo-treated subjects whose panic attacks remitted, with those whose attacks did not remit, and with remitting and nonremitting subjects treated with active medication. Seventy-one subjects who met criteria for panic disorder and for a concurrent depressive diagnosis (major depression, dysthymia, or depressive disorder not otherwise specified [NOS]) were treated with either imipramine (n = 36) or alprazolam (n = 35), and 28 patients were treated with placebo. Sixteen placebo-treated and 40 medication-treated patients were panic free at Week 8. Placebo panic remitters were significantly more ill than active-medication remitters on all measures. In contrast, subjects who achieved panic remission on placebo showed no difference on other clinical measures from active medication nonremitters, and little difference from placebo nonremitters. Results support the need for assessment of all core panic disorder symptoms in determining treatment efficacy and in considerations of response and remission. PMID- 7491380 TI - Skin picking as a symptom of body dysmorphic disorder. AB - Skin picking, traditionally considered a type of "neurotic excoriation," has received little investigation. In particular, its association with body dysmorphic disorder (BDD) has not been studied. A total of 123 subjects with DSM IV BDD were assessed with a variety of instruments; the descriptive characteristics of the 33 subjects who picked their skin were assessed and compared with those of the 90 subjects who did not pick. Of the 14 men and 19 women who engaged in picking behavior, 97 percent had preoccupations involving the skin. Nearly all had experienced impairment in social (97%) or occupational (82%) functioning, 33 percent had attempted suicide, and 67 percent had received dermatologic treatment, which was generally ineffective. Of 35 trials with a serotonin reuptake inhibitor (SRI), 17 (49%) resulted in significant improvement, in contrast to only 4 of 40 (10%) trials with other psychotropics. Subjects who picked were more likely than those who did not pick to have skin preoccupations, to have actual physical defects, to excessively groom and camouflage, and to have received dermatologic treatment. These preliminary findings suggest that compulsive skin picking is an under-recognized problem that commonly occurs as a symptom of BDD, is associated with significant morbidity, and may respond to psychiatric rather than dermatologic treatment. PMID- 7491381 TI - International study of expert judgment on therapeutic use of benzodiazepines and other psychotherapeutic medications: III. Clinical features affecting experts' therapeutic recommendations in anxiety disorders. AB - Our objective was to assemble expert clinical experience and judgment in the treatment of anxiety and related disorders in a systematic, quantitative manner. This article reports on some clinical features apart from diagnosis that may affect choice of strategy in the pharmacotherapy of anxiety disorders. A panel of internationally recognized experts in treating anxiety and depression was constituted by multistage peer nomination. Ninety percent (66 of 73) completed an extensive questionnaire. This report focuses on the expert panel's responses to questions on therapeutic options, based on multi-part case vignettes of several anxiety disorders presenting clinical variations within the same diagnosis. In the presence of higher levels of functional impairment, the experts more often recommended formal psychosocial procedures for adjustment disorder; medication for agoraphobia, social phobia, obsessive-compulsive disorder, and adjustment disorder; and polypharmacy for agoraphobia. Their therapeutic recommendations were not materially affected by chronicity in the case of panic disorder. Under the condition of heavy use of alcohol in the case of generalized anxiety disorder, the experts avoided benzodiazepines in favor of various other medications. In the presence of a serious cardiac conduction defect in the case of obsessive-compulsive disorder, they less often recommended medication. Those who did recommend medication changed their preference from tricyclic antidepressants (clomipramine) to selective serotonin reuptake inhibitors. Under the condition of a more severe precipitating event in the case of adjustment disorder, the experts were more likely to recommend both formal psychosocial intervention and medication.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7491382 TI - Preliminary report: double-blind, placebo-controlled study of fluoxetine in depressed alcoholics. AB - The selective serotonergic agonist fluoxetine has demonstrated efficacy in the treatment of depression and has possible efficacy in the treatment of nondepressed and depressed alcoholics. However, no double-blind, placebo controlled trials with any selective serotonergic medication have been reported in patients who have both major depression and alcoholism. In this study, 21 patients with DSM-III-R diagnoses of both major depressive disorder and alcohol dependence were randomized to fluoxetine or placebo in a 12-week, double-blind, parallel group trial. The patients reported a high level of current episode (52.4%), prior episode (61.9%), and lifetime (76.2%) suicidal behavior. Total alcohol consumption during the 12-week treatment course was significantly lower in the fluoxetine group than the placebo group, after controlling for baseline differences in consumption. The fluoxetine group demonstrated a four-fold greater improvement in depressive symptoms, but this difference did not reach statistical significance in this small sample. These preliminary findings suggest that fluoxetine has potential for treating the excessive alcohol ingestion of depressed alcoholics and may have potential for treating the depressive symptoms of these patients as well. PMID- 7491384 TI - Do white-cell count spikes predict agranulocytosis in clozapine recipients? AB - The use of clozapine is limited by the risk of agranulocytosis. The incidence of agranulocytosis after 1 year was .80 percent in 11,555 patients registered in the Clozaril Patient Management System (CPMS) who received clozapine from February 1990 to April 1991. We noticed a tendency for white-cell counts to spike upward before agranulocytosis occurred. We analyzed the CPMS data to test whether a white-cell count spike at least 15 percent above the previous measurement predicted agranulocytosis within 75 days, with a 21-day lag to allow white-cell counts to decline to levels indicative of agranulocytosis. The occurrence of a spike, entered as a time-dependent covariate in proportional hazards regression, significantly predicted development of agranulocytosis (risk ratio, 3.02; 95% confidence interval, 1.38 to 6.57). Spikes were sensitive though nonspecific predictors, occurring frequently in patients who did not develop agranulocytosis. These results, while exploratory, indicate the potential usefulness of these spikes as guidelines to govern the use of clozapine. PMID- 7491386 TI - Clinical and acoustic measures of the negative syndrome. AB - We studied 12 male schizophrenia patients in a 6-week treatment trial comparing two neuroleptics and placebo. Efficacy in regard to negative symptoms was of interest, and Alphs' Negative Symptom Assessment (NSA) was included. The NSA includes behaviorally anchored ratings such as "prolonged time to response" and "restricted speech output." Audio recordings of interviews with the patients were analyzed acoustically. In general, the behavioral anchors did not prevent the ratings from being influenced by global impressions. There was a strong correlation between each item and the total of other items, but a modest correlation with the relevant behavioral measure. Results suggest that, even with specific and behaviorally anchored items, the clinician's assessment is confounded by global impressions. Treatment effects were greater for the acoustic measures than for the clinical measures. PMID- 7491385 TI - A comparison of European and American dosing regimens of schizophrenic patients on clozapine: efficacy and side effects. AB - We present a comparison of the results from two studies of patients on clozapine. The American study (n = 84; Hillside Hospital, Glen Oaks, NY) and the European study (n = 63; Innsbruck University Clinics, Innsbruck, Austria) both examine efficacy and side effects in schizophrenic patients on this atypical neuroleptic. There is a very substantial difference in the dosing regimen used on both continents and this is reflected in the studies reported. A question with major clinical implications is whether the higher doses commonly used in the United States lead to a better outcome or a different profile of side effects. Outcome as a function of serum concentration will be examined. Results confirm a lower dose, lower clozapine blood level, and a lesser degree of side effects in the Austrian cohort when compared to the American sample. Surprisingly, the clinical efficacy of the lower dosing regimen was superior to the higher dose. Reasons for this anomaly are explored. PMID- 7491387 TI - Prospective cytochrome P450 phenotyping for neuroleptic treatment in dementia. AB - Older patients have more adverse experiences when treated with psychotropics than younger patients. Age-associated physiological changes, such as reductions in hepatic mass and blood flow, magnify genetic and acquired variations in drug metabolism. Cytochrome P450 2D6 (debrisoquine hydroxylase), which is responsible for the metabolism of several antidepressants and neuroleptics, is constitutionally deficient in up to 10 percent of the population. In this study of 45 elderly patients suffering from dementia and treated with perphenazine, 5 patients who were prospectively identified as poor P450 2D6 metabolizers had significantly greater side effects than the 40 extensive metabolizers. PMID- 7491388 TI - Neuroleptic treatment of elderly patients. AB - Data from an ongoing longitudinal study of the development of tardive dyskinesia were analyzed to examine the indications for neuroleptic treatment, continuity of treatment and dosage, and the effects of treatment on ratings of psychopathology and cognitive functioning during the first year of followup. Subjects were 266 elderly patients who had just begun neuroleptic treatment. Patients were predominantly (75%) female; their mean (+/- SD) age was 76.9 (+/- 9.2) years. A psychiatric diagnosis was recorded for 44 percent, primarily major depressive disorder; organic mental syndrome was diagnosed in 65 percent of the patients. Symptom ratings indicated 82 percent of the patients had psychosis, with or without agitation. Haloperidol was prescribed for 68 percent of the patients. Most were on neuroleptic medication continuously during the first month of followup; the average starting dose was 80 mg/day in chlorpromazine equivalent units. Symptom ratings at 1 month and 6 months showed significant improvement from baseline; ratings of dementia were unchanged. PMID- 7491389 TI - Lymphocyte glucocorticoid receptor: predictor of sertraline response in adolescent major depressive disorder (MDD). AB - Major depressive disorder (MDD) in adolescents demonstrates resistance to tricyclic antidepressants and absence of hypercortisolemia. The efficacy of serotonin reuptake inhibitors (SRIs) is uncertain, and response predictors are unavailable. Abnormal fast feedback and negative feedback of the hypothalamic pituitary-adrenal axis implicates a dampened limbic-hippocampal glucocorticoid type II receptor (GCII). We hypothesized that lymphocyte GCII is altered in adolescent MDD and could serve as a marker for response to SRIs. In an open-label study, adolescents (n = 20) meeting DSM-III-R criteria for MDD showed baseline lymphocyte GCII sites per cell (sites/cell) values of 793 +/- 106 versus 2,563 +/ 499 (+/- SEM) for matched controls (n = 18) (t = 3.5; df = 36; p < .001). GCII was bimodally distributed, with SRI responders differing from nonresponders (t = 3.9; df = 14; p < .001). GCII accurately classified 90 percent of sertraline responders and 80 percent of nonresponders. Only SRI responders showed GCII sites/cell upregulated after 6 weeks of treatment (t = 2.1, df = 10; p < .05). PMID- 7491390 TI - The comorbidity of attention-deficit hyperactivity disorder in adolescent mania: potential diagnostic and treatment implications. AB - The frequency of attention-deficit hyperactivity disorder (ADHD) was assessed in 20 adolescents with DSM-III-R bipolar disorder hospitalized for the treatment of acute mania. Thirteen (65%) patients met DSM-III-R criteria for ADHD. Patients who met criteria for both disorders were more likely to be Caucasian and male, and to have mixed rather than pure mania. Patients with ADHD also had a higher mean total Young Mania Rating Scale score and lower serum thyroxine concentrations than patients with bipolar disorder alone. These results suggest that prior histories of ADHD or ADHD symptoms may be common in adolescents hospitalized for mania, and that patients with both disorders may have more severe manic symptoms than patients with mania alone. These findings may have important implications, especially in determining the optimal pharmacologic treatment of patients who meet criteria for both disorders, because standard treatments for ADHD and bipolar disorder have been assumed to exert opposing effects. PMID- 7491383 TI - The utility of diazepam loading in the treatment of alcohol withdrawal among psychiatric inpatients. AB - The aim of this study was to assess the usefulness of an objective rating scale (the modified Clinical Institute Withdrawal Assessment for Alcohol [CIWA-A]) and a diazepam loading dose (20 mg) in the management of alcohol withdrawal among hospitalized psychiatric patients. The sample consisted of 152 adult, hospitalized, psychiatric patients with comorbid alcohol dependence. Of the 152 subjects, 115 (75.7%) completed detoxification with supportive therapy only. Among the 37 patients who required pharmacotherapy (24%), 15 (40.5%) required only one 20-mg dose of diazepam, 14 (38%) required 40 mg to 100 mg, and 8 (21.6%) required doses ranging from 120 mg to 220 mg. There were no complications related to this treatment. The results suggest that the use of the objective rating scale and the diazepam loading dose appears to be safe and effective, and is a useful method for the management of alcohol withdrawal among psychiatric patients. PMID- 7491391 TI - Methylphenidate effects on a laboratory aggression measure in children with ADHD. AB - This study investigated the utility of the Point Subtraction Aggression Paradigm (PSAP), a computerized laboratory analog aggression measure. We measured the effects of methylphenidate (MPH) on aggressive responding in children with a DSM III-R diagnosis of attention deficit hyperactivity disorder (ADHD), who had high ratings on the Child Behavior Checklist Aggression Factor (CBCL; Achenbach 1991). Results are reported for 6 subjects, ages 8-11 years. All were tested at baseline in an unmedicated condition, and after double-blind administration of placebo, 0.3 mg/kg of MPH, and 0.6 mg/kg of MPH. A main effect of decreased aggressive responding with MPH was found, with a dose-related change by repeated measures ANOVA (F = 6.59, df = 2.10, p = .014). Post-hoc analysis with the Tukey HSD indicated that only the 0.6 mg/kg was significantly (p < .05) different than placebo. Implications for use of the PSAP in future investigations of medications in aggressive children are discussed. PMID- 7491392 TI - Neuropsychological effects of amphetamine may correlate with personality characteristics. AB - Although the stimulant properties of amphetamine are well established, its effects on cognitive test performance in unfatigued normal adults are poorly documented. Seventeen healthy individuals received a single oral dose of dextroamphetamine (0.25 mg/kg) in a double-blind, placebo-controlled, crossover study. Neurocognitive tests, motor tests, and behavioral observations were performed. Personality information, based on the Tridimensional Personality Questionnaire (TPQ) was also gathered to explore a relationship between personality factors and response to the stimulant. With the exception of two measures of reaction time, there were no overall changes in performance on measures of memory or other cognitive functions. There was decreased reaction time on the continuous performance task (CPT) and increased accuracy of performance under minimal delay conditions in the spatial delay response task while subjects were receiving amphetamine. In addition, the novelty-seeking subscale was found to correlate with a measure of verbal memory. Individuals with higher scores on the novelty-seeking scale deteriorated under amphetamine, while those who had lower scores improved. These results suggest that some cognitive abilities of persons who may have relatively high dopaminergic tone are disrupted by amphetamine, while those with relatively low dopaminergic tone may have their performance enhanced. PMID- 7491393 TI - Haloperidol-related dyskinesias and pre- and perinatal complications in autistic children. AB - It has been hypothesized that an association exists between pre- and perinatal complications, central nervous system (CNS) dysfunction, and the development of tardive (TD) and withdrawal dyskinesias (WD). We assessed the relationship between pre- and perinatal complications and TD/WD in a sample of 118 children with autism who participated in an ongoing long-term prospective study of the efficacy and safety of haloperidol. The mean total Rochester Research Obstetrical Scale (ROS) score was significantly higher for children who developed TD/WD compared to those who did not (p = .007) as was the mean score of the ROS Delivery Scale (p = .002). Anesthesia during delivery was more frequent in children who developed TD/WD (25 of 40, 62.5%) than in those who did not (30 of 78, 38.5%) (p = .019). The ROS Not Vertex Presentation item and TD/WD were associated only in females (p = .019). Six of 7 males with short labor developed TD/WD (p = .007). ROS scores did not differ significantly as a function of gender or socio-economic status (SES). Pre- and perinatal complications appear to be related to the development of TD/WD in this sample of children. PMID- 7491395 TI - The lithium test dose prediction method in aggressive children. AB - Cooper and associates (1973) developed a method of ascertaining the lithium dosage required to attain a therapeutic serum level of 0.6 to 1.2 mEq/L. However, reports about the safety and accuracy of their method in children are limited (Geller & Fetner 1989). This study relates our experience with using this method in children. Subjects were 16 conduct-disordered children (13 males, 3 females), ages 8.97 to 17.14 years (mean, 12.73 +/- 2.12), who were treated with lithium to decrease aggressive behavior. Following a lithium 600-mg loading dose, a 24-hour serum lithium level was drawn from which a "predicted" lithium dosage was established. These dosages ranged from 600 to 1,800 mg/day (mean, 1,312.5 +/- 450) and the corresponding serum lithium levels at steady-state ranged from 0.58 to 1.13 mEq/L (mean, 0.87 +/- 0.15). No severe side effects were encountered. This suggests that the method is safe and useful for predicting lithium dosages in children. PMID- 7491394 TI - A comparison of live and videotape ratings: clomipramine and haloperidol in autism. AB - This study compared live ratings with ratings of videotapes and compared response to clomipramine with response to haloperidol in 8 subjects, mean age 5.62 years, who met criteria for autism. They were consecutive admissions to a pilot study of clomipramine (n = 4) or a double-blind, placebo controlled study of haloperidol (n = 4). Live ratings were performed by two raters at the end of the pre treatment placebo baseline period and at the end of the drug treatment period on the CPRS and the CGI and were videotaped. Employing the same instruments, these videotapes were rated by two raters who did not know the subjects and were blind to study design, treatment, and study phase. Ratings of videotapes significantly differed from live ratings. A treatment effect for haloperidol was detected only on live ratings and not on ratings of videotapes. No treatment effect was detected for clomipramine in either live or videotape ratings. PMID- 7491396 TI - Schizophrenia in hospitalized adolescents: clinical diagnosis, DSM-III-R, DSM-IV, and ICD-10 criteria. AB - The DSM-III-R, DSM-IV, and ICD-10 criteria were tested in hospitalized adolescents diagnosed with schizophrenia and other psychotic disorders employing a retrospective chart review. The charts of 111 patients, ages 11 to 17 years, representing consecutive admissions to Bellevue Hospital Center over a period of 18 months were reviewed. Thirty patients had a clinical diagnosis of schizophrenia or other psychotic disorders on admission and were selected for the present study. The 30 patients were independently rediagnosed using the criteria of DSM-III-R, DSM-IV, and ICD-10. Schizophrenia was diagnosed clinically in 6 patients, 10 met DSM-III-R criteria, 9 met DSM-IV criteria, and 12 met the ICD-10 criteria. Agreement for schizophrenia was high across the diagnostic systems. The clinical diagnosis of psychotic disorder, not otherwise specified (NOS) was overinclusive. The mean number of psychotic symptoms per subject was 2.66 (range 1-6). We found no significant relationship between the frequency or type of symptoms and the age or gender of subjects. PMID- 7491397 TI - Comparative analysis of three PMS assessment instruments--the identification of premenstrual syndrome with core symptoms. AB - Premenstrual syndrome (PMS) is a commonly encountered complaint among women. This study compared the PMS Diary (PMSD), which measures core menstrual symptoms (negative affect, water retention, and pain symptoms), with two commonly used self-rating forms, the Menstrual Distress Questionnaire (MDQ) and the Daily Rating Form (DRF). Thirty-seven premenopausal women with documented PMS completed the forms. A multitrait-multimethod analysis was performed to determine overall agreement and reliability. The three instruments and their component scale scores had strong internal consistency. Correlations between overall scores and between scales measuring similar constructs were strong. Correlations were 0.77 between PMSD and MDQ; 0.67 between PMSD and DRF; and 0.81 between MDQ and DRF. The PMSD performs as effectively as more extensive questionnaires in measuring symptoms in women with PMS. The PMS Diary is a concise yet reliable and valid instrument that can be easily administered in ambulatory care and longitudinal research. PMID- 7491398 TI - Assessment of paroxetine-induced sexual dysfunction using the Changes in Sexual Functioning Questionnaire. AB - The Changes in Sexual Functioning Questionnaire (see Appendix) was developed as a brief clinical and research instrument to measure sexual function changes accompanying illness or the administration of medications. In a pilot study of patients with major depression being treated with paroxetine, the instrument was useful in delineating three groups of patients: patients who reported increased libido without sexual dysfunction, patients who developed anorgasmia with possible spontaneous resolution (medication side effect and development of tolerance), and patients with long-term sexual dysfunction unaffected by the medication. Change scores accurately differentiated global elements, changes associated with specific factors of sexual response, and drug-specific side effects. All patients had a therapeutic response to treatment with experiences of sexual functioning that could be categorized into one of the three groups. PMID- 7491399 TI - The SDDS-PC: a diagnostic aid for multiple mental disorders in primary care. AB - The Symptom Driven Diagnostic System for Primary Care (SDDS-PC) is a new computerized clinical procedure to assist primary care physicians in diagnosing mental disorders during the course of routine practice. It has three components: (1) a 5-minute patient-administered 16-item screening questionnaire, (2) six 5 minute physician-administered diagnostic interview modules based on DSM-III-R criteria, and (3) a longitudinal tracking form. The SDDS-PC covers five disorders (major depression, panic disorder, alcohol abuse or dependence, generalized anxiety disorder, and obsessive compulsive disorder) as well as suicidal ideation. Patients who screen positive for a disorder receive the corresponding diagnostic interview module. Patients who meet mental disorder criteria on the diagnostic interview module are then followed with the longitudinal tracking form. Minor or subsyndromal conditions are also addressed at the physician's discretion. This article describes the development of SDDS-PC and summarizes results from two studies which involved comparisons between the SDDS-PC and independently administered full-length structured diagnostic interviews. PMID- 7491400 TI - Cues to the assessment of affects and moods: speech fluency and pausing. AB - Clinicians' rating of the negative syndrome of schizophrenia are highly correlated with the rate and duration of the patient's pauses. To test the degree to which pausing provides the cue for clinicians' impressions we selected audio recordings of interviews with schizophrenic patients who showed moderate levels of negative signs and, through the use of a computerized digital editor, transformed the recordings by either doubling or halving the duration of all of the patient's pauses. Other aspects of the patients' speech were unchanged. Ratings of flat affect were changed by the editing: reduced pause samples were seen as less flat and increased pause samples as flatter. Ratings of mood were not significantly changed. These results suggest that neuropsychological factors that interfere with fluent speech may play a role in the pathogenesis of the negative syndrome. In addition, the edited tapes provide helpful materials for training clinicians to assess moods and affects. PMID- 7491402 TI - Pitfalls in presenting and interpreting clinical trial data. AB - Information generated by a clinical trial, when conveyed to health professionals and prospective patients, is affected by the original design of the trial and by the manner in which the results are presented. One problem in study design is the management of comparison groups in randomized assignments. When a comparison group is treated with an accepted standard compound, the chosen standard drug may be one that is associated with more side effects and complications than later modifications of the standard. Inadequate dosing of the comparison group can inflate the relative effect size of the experimental compound. Choosing a standard with a verifiable dose reference range can avoid this pitfall. In reporting results, relative score changes on a rating scale are meaningless without reference to an absolute value reflecting a clinically relevant degree of remission. The validity of rating instruments chosen must be judged in the context of the specific population to which it is applied. In the reporting of effects, the emphasis on significance of differences may obscure the critical distinction between statistical significance and clinical relevance, and graphs can appear to overstate a change over time by truncating the ordinate axis. PMID- 7491401 TI - Psychometric characteristics of the Wender Utah Rating Scale (WURS): reliability and factor structure for men and women. AB - The goals of this study were to examine the factor structure of the Wender Utah Rating Scale (WURS), to evaluate potential gender differences in factor composition, and to assess the reliability of the scale. The WURS was completed by 310 fathers and 305 mothers of children referred for evaluation of Attention Deficit Hyperactivity Disorder (ADHD). For males, a five-factor solution (Conduct Problems, Learning Problems, Stress Intolerance, Attention Problems, Poor Social Skills/Awkward) accounted for 72 percent of the variance. There was also a five factor solution for females (Dysphoria, Impulsive/Conduct, Learning Problems, Attention and Organizational Problems, Unpopular) which accounted for 71 percent of the variance. Symptoms of inattention and impulsivity loaded on separate factors for both men and for women. Test-retest reliability was examined with a different sample of 57 adults who completed the WURS on two separate occasions, 1 month apart. The WURS demonstrated satisfactory internal consistency and temporal stability, and it may be a useful tool for the study of ADHD in adults. PMID- 7491403 TI - Venlafaxine: measuring the onset of antidepressant action. AB - Venlafaxine, a new antidepressant, inhibits reuptake of norepinephrine and serotonin without appreciable effects on histaminergic, alpha-adrenergic, or cholinergic systems. Pharmacologically the drug is unique: the half-life is short and it exerts both rapid and prolonged beta-adrenergic desensitization after single doses in a rodent model. Venlafaxine has been thought to possess a rapid onset of clinical antidepressant action. Accordingly, two clinical studies in which moderate amounts of venlafaxine were given aggressively were reviewed to examine aspects of the drug's onset of action. Three statistical methodologies were employed--traditional analysis of depression scale scores, pattern analysis based on timing and persistence of response, and survival analysis of sustained response. All three methods showed venlafaxine to have significant effects early in the course of therapy. In addition, venlafaxine is the first drug to meet criteria for early onset using the pattern analysis methodology. Depressed patients aggressively treated with venlafaxine show significant benefit on or before Day 7 of treatment using traditional methods of analysis as well as survival analysis of sustained response. PMID- 7491404 TI - Down-regulation of PKC alpha by lithium in vitro. AB - In recent years, research aimed at the elucidation of lithium's molecular mechanisms has focused on signal transduction pathways. This research has demonstrated that lithium has multiple effects on the phosphoinositide turnover signaling system. We have previously demonstrated that chronic (but not acute) in vitro exposure of HL60 cells to 1 mM lithium reduces both receptor and phorbol ester-mediated Na+/H+ activity without affecting agonist-induced increases in intracellular Ca2+ or phosphoinositide breakdown, findings which suggest an attenuation of protein kinase C (PKC) function. The present study sought to measure the in vitro effects of lithium on PKC more directly and demonstrated that 5-day in vitro exposure of HL60 cells to either 1 mM or 10 mM lithium chloride dramatically reduces PKC alpha in both cytosolic and membrane fractions. Given the critical role of PKC in regulating neuronal signal transduction, these effects may play a major role in lithium's mood-stabilizing effects. PMID- 7491405 TI - Postpositivist methodology and clinical education. AB - This article presents an alternative research methodology to improve clinical teaching and learning. The author describes the characteristics of the postpositivist paradigm and discusses how adoption of this paradigm and its associated research methods can benefit the clinical practicum in radiography. PMID- 7491408 TI - Monoclonal antibodies: the promise and the reality. AB - Monoclonal antibodies, or "MoAbs," have revolutionized clinical approaches to diagnostic imaging and therapy of many diseases. The use of MoAbs for diagnosing and treating cancer has been especially promising. However, the full potential of these "magic bullets" has yet to be realized. This article examines the current and potential uses of MoAbs, describes problems with the technology and looks at potential solutions. Along with descriptions of how MoAbs are made and prepared for use in the clinic, the article provides examples of the ways in which MoAbs can be used to complement and expand the information obtained from standard diagnostic imaging modalities. Specific examples of the use of monoclonal antibodies for treating cancer and other diseases also are provided. PMID- 7491414 TI - One for the archives. PMID- 7491412 TI - Clinical applications of spiral CTA. PMID- 7491409 TI - Therapeutic uses of radiation. PMID- 7491413 TI - The Internet's role in education. PMID- 7491407 TI - MR imaging of macroadenomas. AB - Pituitary adenomas account for about 15% of all intracranial neoplasms. Patients with symptoms and signs of pituitary axis dysfunction or visual field deficits frequently are referred for diagnostic imaging procedures. Magnetic resonance imaging is the modality of choice for evaluating the sellar and juxta-sellar region. MRI provides multiplanar images with superior tissue contrast differentiation. This article describes how MRI is used to image pituitary macroadenomas. PMID- 7491411 TI - From another place and time. PMID- 7491410 TI - Stereolithography and the Iceman. PMID- 7491415 TI - [Use of molecular biology in epidemiology in rabies]. PMID- 7491406 TI - Critical-thinking abilities of radiologic science students. AB - This article reports the results of a survey conducted to determine whether critical thinking abilities differ among radiologic technology students educated at the baccalaureate, associate degree and certificate levels. The sample population was 357 students enrolled in radiologic technology programs in Louisiana, Missouri and Oklahoma who completed the Watson-Glaser Critical Thinking Appraisal. Results showed a significant difference in critical thinking abilities at the three educational levels. Baccalaureate students scored significantly higher in critical thinking abilities than associate and certificate students, and certificate students scored significantly higher than associate degree students. PMID- 7491416 TI - [Animal sources of rabies infection in Poland in 1985-1994. Epizootiological tendencies and dynamics of the focus before the introduction of mass wildlife oral antirabies vaccination]. AB - The purpose of this paper is to collect (four tables), evaluate and map (two figures) the numbers and dissemination of rabies among animal species in Poland during the last ten years. This information is required for planning, organizing and implementing of antirabies wildlife oral vaccination and for evaluation of their effectiveness. PMID- 7491417 TI - [Vibrio cholera (Vibrio cholerae non-01) isolated in Poland from the Bug river]. AB - Non-01 cholera vibrios for the first time has been isolated from freshwater in Poland. In October 1994 during bacteriological examination of Bug water by a two step enrichment method in alkaline peptone water at pH 8.6 and TCBS. Repeated examination in November, December and January in the same and other locations among the polish-ukrainian borderline revealed persistence of V. cholerae in the river; 22 strains were isolated. All 22 strains were identical to V. cholerae E1 Tor 01 Ogawa reference strain except to antigens 0. The strains isolated from water were no agglutinable by standard 01 serum neither by 0139 serum. Determination of their antigens 0 is under investigations. They might be pathogenic for people as it could be judged by their high resistance to bile, proteolytic and haemolytic activity. No cholera-like cases were notified on the polish side of the river. PMID- 7491418 TI - [The role of ticks in the epidemiology of Lyme borreliosis]. AB - In this paper the current knowledge of the natural circulation of Borrelia burgdorferi, the etiologic agent of Lyme borreliosis, and its relationships both to hard ticks (Ixodidae) serving as vectors, and to certain vertebrates which function as tick hosts and pathogen reservoirs is reviewed. The modes of transmission of borreliae (by salivation, regurgitation, defecation) by ticks to their animal hosts and men as well as prophylactic measures against tick-bites are also discussed. PMID- 7491419 TI - [Seasonal prevalence of antibodies against Borrelia burgdorferi in Bialowiez inhabitants]. AB - The aim of study was evaluation of seasonal prevalence of antibodies against Borrelia burgdorferi in Bialowieza area, that is Lyme borreliosis endemic region. IgM and IgG antibodies against 41-kDa antigen of Borrelia burgdorferi B31 strain were investigated with ELISA in sera of 300 Bialowieza inhabitants between September 1993 and May 1994. Moreover in 40 asymptomatic persons, that were seropositive in September 1993, additional control was performed in May 1994. The highest prevalence of IgM antibodies was observed in September 1993 (43.4%), with decrease during winter and spring (down to 15.7% in April) and accompanying the highest prevalence of IgG antibodies (17.7%) in April 1994). From 33 persons, that revealed IgM antibodies in September 1993, 3 (9%) were still seropositive, 4 (12%) cleared IgM antibodies and became IgG-seropositive, and remained 26 (79%) had no antibodies. Specific IgG antibodies found in 7 persons in September, were still present in 4. Obtained results indicate necessity of taking into consideration seasonal dynamics, when results of seroconversion against Borrelia burgdorferi between different areas and different populations are compared. PMID- 7491420 TI - [A comparison of serologic ELISA tests from BIOMEDICA Ges. M.B.H. and VIRO-Immun Labor Diagnostica GMBH in the diagnosis of Lyme disease (introduction)]. AB - The purpose of this work was to evaluate detection of antibodies IgM and IgG against B. burgdorferi. We examined presence of IgM in 96 serum samples from patients suspected of Lyme disease. We looked for IgG in serum samples of 48 forest workers who submitted frequent tick bites in last two years. Among 96 patients whose sera were examined we had positive results in both tests in 5 cases and in 25 cases positive results were achieved only in test II. In 48 serum samples which we examined for presence of IgG in 2 cases positive results were in both tests, in 1 case positive in test II and negative in test I in 3 cases positive result in test I and negative in test II. According to these results we drew following initial conclusions: 1) obtained results testify better sensitivity and specificity of test II which consists of recombinant antigens of Borrelia burgdorferi, 2) no essential difference was shown in detection IgG among two used tests. PMID- 7491421 TI - [Subacute sclerosing panencephalitis in Poland in the years 1990-1993. Stage V of epidemiologic studies]. AB - The authors summarize the results of the fifth stage of epidemiological investigation on SSPE. The first stage covered the years 1971-1976, the next stages covered the periods: 1977-1983; 1984-1986; 1987-1989. In the period 1990 1993 data on 95 patients with SSPE were obtained. Morbidity rates continued to show a decreasing tendency (from 0.8 to 0.6 per 1,000,000 population) in the years 1990-1993, being lower the median morbidity (amounting to 0.98) in 1985 1989. Morbidity rates continued to shift towards older age groups: while the peak morbidity (25.7%) in the years 1990-1992 was noted in the 14-15 age range, in 1993 the peak (28.6%) was found in the age group of 16 to 19 years. PMID- 7491422 TI - [The role of Haemophilus influenzae in epidemiology of respiratory infection and meningitis]. AB - Epidemiological situation of respiratory infection and meningitidis due to Haemophilus influenzae in Poland and in the world were described. We discussed carrier state of Haemophilus and risk factors for invasive H. influenzae disease. PMID- 7491423 TI - [Properties of S. aureus and H. influenzae strains residing in the throats of children]. AB - Strains of S. aureus and H. influenzae which were isolated from the throats of 41 children suffering from successive acute pharyngitis and tonsillitis were compared. All strains which were isolated at least twice, during acute occurrences, or more from the throat of each child, were traced by: phago-typing, biochemical properties, antibiogram and ability to produce beta-lactamases (S. aureus), or by: biotyping, serotyping, ability to produce beta-lactamases and antibiogram (H. influenzae). Based on these investigations it was concluded that the bacterial strains among the examined children had often changed. It was concluded that only a few children were a carriers of the same strain of S. aureus. PMID- 7491424 TI - [Analysis of the occurrence of intestinal parasites among children in first grade in elementary school in Poland in the school year 1992/3]. AB - In 1992/93 there were reexamined in Poland 10% of the population of children aged 7 years for the presence of intestinal parasites, continuing country-wide operation launched in 1988. The examination included 60,288 children, 38,003 inhabiting towns and 22,285 from the country. Among the examined in each voivodeship the numbers of children from towns and country were proportional to the actual number of children living in the two localities. The methods included examination of direct faecal smear in 0.85% NaCl and in Lugol's iodine, zinc sulphate flotation, decantation and cellophane swabs according to NIH. The parasites encountered in order of decreasing frequency comprised: E. vermicularis, G. intestinalis, Entamoeba coli, A. lumbricoides and T. trichiura. The highest number of infected children was found in the Biala Podlaska district. The number of infected among children inhabiting country was significantly higher than among those from the towns. The obtained results have confirmed hitherto existing views that in Poland the epidemiological situation of infections with intestinal parasites is steadily improving. PMID- 7491425 TI - [The occurrence of viral respiratory tract infections in small children in the season 1993-1994 (with reference to the role of breast feeding)]. AB - In the period October 1993 to June 1994, 231 hospitalized children under 2 years of age were examined by immunofluorescence for respiratory viruses. In 114 (49.9%) cases viral etiology was established. The data were compared with results obtained from the season 1992-93. RS virus was predominant as in the previous season but infections of parainfluenzae type 3 virus were rather rare (p < 0.005). Mixed and adenovirus (p < 0.02%) infections were more frequent than in the past season. A breast-fed children were statistical more rarely infected by respiratory viruses (p < 0.05), (RS virus-p < 0.1), than artificial-fed ones. PMID- 7491426 TI - [Beta 2-microglobulin in serum of patients with infectious mononucleosis]. AB - Serum beta 2-microglobulin concentrations were assayed in twenty patients with infectious mononucleosis. Pairs of sera taken from each patient in the first week of hospitalisation and two weeks later were tested. There was a significant increase of beta 2-microglobulin levels in these patients compared to the control group. A raised serum beta 2-microglobulin concentration persists during the convalescent period. PMID- 7491427 TI - [Characterization of Candida sp strains isolated from different clinical specimens in the years 1989-1993]. AB - 420 Candida sp. strains, which were isolated from the following specimens: vaginal secretions, throat swabs, urine, surgical specimen, nail specimen, sputum, semen, bile, skin, swab, pus were examined. On the basis of morphological and biochemical examinations it was established that in the above mentioned specimens are presented different species of Candida genus. Most often were isolated C. albicans, C. krusei, C. tropicalis. Among 317 C. albicans strains, 306 belonged to serotype A, 11 strains belonged to serotype B. The determination of enzymatic properties made possible the differentiation of strains C. albicans species from other species, and made it possible to define the biotypes. Sensitivity tests for examined strains made the characterization more particular. PMID- 7491428 TI - [Infection with HCV in patients hospitalized for various causes. I. Prevalence of anti-HCV antibodies in selected groups of patients]. AB - Using the third generation ELISA test UBI HCV (Organon Teknika) we studied the prevalence of anti HCV antibodies among 980 patients (hospitalized of various diseases) in 4 hospitals in district Lublin. The results indicated that 22.3% of patients were positive for anti HCV. The presence of anti HCV in the high-risk group was higher, in hemophiliacs-59% and in hemodialysis patients-57%. The prevalence of anti HCV is closely correlated with age (19.2% in children and 24.4% in adults). This difference was statistically significant (p < 0.001). As nosocomial infections was probably 70% of cases. PMID- 7491429 TI - [Concurrence of HBs antigen and anti-HBs antibodies in children infected with HBV]. AB - Two cases of concurrence of HBs antigen and antibodies in boys, who acquired HBV infection within the first two years of life are described. Authors discuss possible explanation of that fact. PMID- 7491430 TI - [Vaccination against varicella]. AB - Varicella is generally a mild infections disease of children age. It may be danger among others for people especially children with biological or iatrogenic immunosuppression for instance with diagnosis of leukemia, other neoplastic diseases and with transplantation of tissues. Live, attenuated vaccine was produced in Biken Institute in 1974. It was found as immunogenic, effective, small reactogenic and stabile. The price of vaccine is very high (one dose--74 DM). Vaccination of children against varicella with immunosuppression in the course of leukemia or other diseases performed in hospitals or clinics where they are treated is advisable actually in Poland. PMID- 7491431 TI - [Acellular pertussis vaccine (Pa)]. AB - Vaccination by whole-cell pertussis vaccine (Pw) since forties years was followed by decreasing of morbidity and especially mortality caused by pertussis. High reactogenicity of this vaccine (Pw) was the reason trials undertaken to search new type of less reactogenic vaccine. Acellular pertussis vaccine (Pa) was found as effective, safe, small reactogenic. It may be especially used as booster doses for older children and even for adults people. Actual pertussis infection among adults are suspected as the source infection for children. Just now in some countries acellular vaccine start to be used for ground vaccination of children. Increasing of morbidity of pertussis was noted in some provinces in Poland in last time. Acellular pertussis vaccine may be introduced as booster dose for 6-7 years old children, if epidemiological situation of pertussis among school-age children would be not satisfied in future. PMID- 7491432 TI - [Prospective epidemiologic study on respiratory diseases in children in Cracow. Pilot study]. AB - After introducing the main purpose of the project on the outdoor and indoor air quality and children's health in Cracow, the paper presents the results of the pilot study carried out in the higher polluted city area (suspended particulate matter: 51.5-74.5 micrograms/m3/year; SO2: 58.4-73.8 micrograms/m33/year) and in the less polluted area (suspended particulate matter: 31.7 micrograms/m3/year; SO2: 36.1 microgram/m3/year). The pilot study covered 171 children. There was an excess of chronic respiratory symptoms in children from the higher polluted area, but only the frequency of shortness of breath in boys and attacks of shortness of breath with wheezing in girls were significantly higher in those living in the polluted area. Spirometric indices were significantly lower in both boys and girls in the residence area with the higher air pollution level. The data on prevalence of respiratory diseases in the pilot study will be used to estimate the size of the sample needed in the other stages of the prospective study. PMID- 7491433 TI - [Incidence of endemic goiter--environmental factors and iodine prophylaxis in Southern Poland]. AB - In Southern Poland a well defined endemic goiter area has been known since 19th century. This region was characterized by low level of iodine in the environment. Iodine prophylaxis in Carpathian endemic area was introduced as early as 1935 at the level of 5 mg/kg of table salt. The goiter prophylaxis programme was interrupted during the Second World War. After the war in the years 1945-1946 epidemiological studies held in Southern Poland paid attention to high percentage of goiter in the population. As the result, potassium iodide salt supplementation started in 1947, but goiter incidence in subsequent years was still noted. As an effect of these observations, since 1956 potassium iodide salt supplementation was increased from 5 to 12 mg/kg of salt--in Carpathian and Subcarpathian regions. During over 30 years of iodide prophylaxis the severe symptoms of iodine deficiency disorders disappeared and endemic goiter in Southern Poland became mild. In 1980 the supplementation of kitchen salt was discontinued due to technical reasons. In 1985 an increase of goiter incidence in adults and children, as well as iodide deficiency disorders in newborns was noted. That is why in 1986 the salt supplementation was reintroduced at the level of 20 mg KJ/kg of kitchen salt. The potassium iodide content assessment in salt for human consumption in Krakow region was carried out in the years 1989-1994. The percentage of salt samples with the proper content of potassium iodide was 50, so it meant that the technological process of salt iodination was done incorrectly. Medical investigations are necessary to establish the optimal preventive dose of KJ.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7491436 TI - [Ventilatory assistance in acute exacerbations of chronic respiratory insufficiency in adults (with the exclusion of neuromuscular pathology and weaning). 13th Consensus Conference in Resuscitation and Emergency Medicine. 2 December 1994, Strasbourg]. PMID- 7491434 TI - [Indications for respiratory function tests (RFT). The role of RFT and blood gases in the monitoring of asthma and chronic obstructive bronchopnumopathies]. PMID- 7491438 TI - [Clinical case of the month. Occupational rehabilitation of the diabetic patient]. PMID- 7491437 TI - [Pharmaclinics--how I treat... Crohn disease]. PMID- 7491435 TI - [Tuberculosis in France: how can the current epidemic be stopped? Conference of Experts on Tuberculosis, 15 December 1994, Paris]. PMID- 7491439 TI - [Current therapies 1990-1995 in infectious diseases]. PMID- 7491441 TI - [Diaper dermatitis]. PMID- 7491440 TI - [Vascular endothelium and vasomotor tonus]. PMID- 7491442 TI - [Sickle cell anemia: its clinical implications in dental care]. PMID- 7491444 TI - [How I examine... Anemia (1)]. PMID- 7491443 TI - [Venous allografts for salvage of lower limbs: hope and reality]. PMID- 7491445 TI - [Pharmaclinics. Drug of the month. Captopril (Capoten) in diabetic nephropathies]. PMID- 7491446 TI - [Cardiovascular pharmacotherapy in old age: How beneficial is it? What does it cost?]. AB - Because of their high morbidity and mortality, elderly people benefit most from drugs with a proven effect on disease complications and premature death. Thiazides, beta adrenergic blocking drugs, oral anticoagulants and aspirin are drugs that satisfy such criteria. These medications are comparatively well tolerated and cheap. They should not be neglected in favour of newer drugs with a less complete performance record. PMID- 7491448 TI - [Cardiovascular diseases, cognitive abilities and quality of life in old age]. AB - Dementia is the most important disorder endangering and impairing quality of life in old age. Cardiovascular diseases as well as cardiovascular risk factors raise the risk for dementia; therefore, a targeted primary and secondary prevention strategy is indicated, also in view of the maintenance of autonomy and quality of life. Consequent primary and secondary prevention comprises adequate therapy of risk factors, including pharmacotherapy, if necessary. Important general measures are regular physical and mental training, complemented by an alimentation with a favorable composition of lipids and rich in micronutrients (in particular antioxidants). PMID- 7491449 TI - [Sports and rehabilitation in old age]. AB - Cardiac rehabilitation of older patients aims at restoring physical aptitude as well as possible and at preserving independence. Prognostic aspects are less important than symptomatic relief. A formal rehabilitation program is indicated in adjunction to revascularization procedures and in cases, where a closer medical supervision is warranted. Exercise guidelines for persons older than 65 years should include prescriptions for endurance training of moderate intensity and an exercise program to enhance flexibility and coordination. Weight training at 40 to 60% of maximal voluntary contraction to support muscle-strength is feasible and useful even in frail persons, whereas sports with high impact of technical skills and speed are not advisable. Such training prescriptions will allow to maintain physiological and psychological function and will thus enhance the quality of life of older patients. PMID- 7491450 TI - [Diabetic nephropathy: significance of microalbuminuria and proteinuria in Type I and Type II diabetes mellitus]. AB - Diabetic nephropathy is a progressive renal disease and represents a serious late complication of diabetes. There are familial clustering and huge ethnic differences in the occurrence of diabetic nephropathy, which point to a genetic predisposition. Diabetic nephropathy is defined by persistent albuminuria (albumin excretion rate [AER] > 300 mg/day), declining glomerular filtration rate and rising blood pressure. Several years of incipient nephropathy, characterized by worsening microalbuminuria (AER 30 to 300 mg/day or 20 to 200 micrograms/min), which is Albustix-negative and detectable by special assays only, are followed by established nephropathy. The natural history of nephropathy differs between insulin-dependent (IDDM) and non-insulin-dependent (NIDDM) diabetes mellitus. In IDDM, nephropathy develops in 30 to 40% of cases. The incidence peaks after 15 to 16 years of diabetes. In NIDDM, estimates of prevalence range from 15 to 20%, and nephropathy often supervenes after a shorter known duration of diabetes than in IDDM. GFR is often increased above normal (hyperfiltration) from the onset of IDDM due to increased renal blood flow, glomerular capillary hypertension and increased filtration surface. The glomeruli are hypertrophied and the kidneys enlarged. In both IDDM and NIDDM, GFR begins to decline irreversibly, when AER has risen to 100 to 300 mg/day at an average rate of 10 ml/min. per year. This is due to progressive reduction of the filtration surface area through mesangial expansion. Serum creatinine levels begin to rise when GFR falls below 50 ml/min, and then end-stage renal failure follows after an average of five years.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7491451 TI - [Heart failure in elderly patients]. AB - The incidence and prevalence of congestive heart failure increase exponentially with advancing age. Congestive heart failure in the elderly is characterized by a multifactorial etiology, a high proportion of accompanying degenerative changes of the cardiovascular system and age-specific problems regarding diagnosis and treatment. The treatment strategy is the same as in younger patients, but the higher incidence of adverse effects and complications demands special awareness. The majority of decompensations leading to hospitalization are precipitated by insufficient compliance in life style change and drug intake. PMID- 7491447 TI - [Hypertension in the elderly: definition, indications for treatment and clarification]. AB - Hypertension is the single most potent common and remediable risk factor for cardiovascular morbidity and mortality in the population over age 65. Data from newer intervention studies show that it is clearly beneficial to treat elderly patients with either both systolic and diastolic hypertension or with isolated systolic hypertension. The indication to start treatment in a given patient depends on blood pressure values, age and eventual concomitant disease. In the evaluation of the elderly hypertensive patient, multiple blood pressure measurements in the standing and lying patient should be taken. In the case of suspected carotid artery stenosis, a Doppler ultrasound is indicated before starting antihypertensive therapy. Finally, if refractory hypertension or kidney failure develop in an elderly patient, renal artery stenosis should be considered. PMID- 7491453 TI - [Extrahospital approach to various emergency situations]. PMID- 7491454 TI - [Suspicion of unstable angina and myocardial infarct. Evaluation and prehospital management by the practitioner]. AB - When confronted with chest pain, one should above all consider unstable infarction or unstable angina pectoris, because these events necessitate immediate hospitalization in view of intravenous thrombolysis. In this article, published works on estimates for the probability of acute myocardial infarction based on history and ECG are summarized. The different decision-making processes practitioners may become confronted with as well as some diagnostic and therapeutic recommendations are introduced. PMID- 7491452 TI - [Treatment of hypertension in elderly patients]. AB - Today indications for treatment of hypertension in older age are no longer contested, even if only systolic pressure is raised. The choice of the antihypertensive drug for elderly patients, however, depends on concomitant diseases. The diversity of the latter demands a judicious, individualized use of the available substances (diuretics, beta-blockers, calcium antagonists and ACE inhibitors). In elderly patients as well, a marked reduction of the cardiovascular risk is achievable by pharmacotherapy. So far, however, such results have only been reported for the use of diuretics and beta-blockers, in purely systolic hypertension by diuretics exclusively; however smaller studies, suggest that comparably favorable results may be obtained with calcium antagonists. PMID- 7491455 TI - [Emergency and hypertensive crisis: definition and treatment]. AB - The opinions on how fast high blood pressure values should be lowered in emergencies have changed over the last years. It seems now more and more reasonable to lower high blood pressure values over several hours rather than several minutes. In some cases, however, a rapid reduction of elevated blood pressure may be necessary. In these cases the risks of a rapid decrease of blood pressure with respect to cerebrovascular circulation have to be considered. PMID- 7491458 TI - [Horton's disease: ophthalmological and neurological complication]. AB - We report the case of a 74-year-old women admitted to the hospital because of rapidly developing diplopia. The admission anamnesis revealed fronto-temporal headaches and pain in the maxilla for three days. A cerebral cT scan was normal, and a corticosteroid therapy was initiated. Diagnosis of Horton's syndrome was confirmed by the good response to treatment and by biopsy of the temporal artery. Different ocular and neurologic complications of Horton's disease, their clinical appearances and therapeutic measures are discussed by the authors. PMID- 7491457 TI - [Emergencies in diabetic patients]. AB - In this article emergencies are presented from the view of the practitioner. At first, hypoglycemias caused by insulin and sulfonylurea are analyzed, emphasizing in particular the necessity for surveillance of hypoglycemias due to sulfonylurea drugs in hospital. Hyperosmolar and ketoacidotic decompensations are reviewed and the limits of the ambulatory care stressed. Finally, some less classical but nevertheless frequent situations like podiatric, ophthalmologic and neurologic urgencies of diabetics are described. PMID- 7491459 TI - [Anaphylactic shock]. AB - Anaphylactic shock is an unexpected, sudden and sometimes deadly event that attacks the patient in 75% of the cases without pre-existent history of allergy. In general, drugs, hymenopteric poisons and nutrients (according to the recent concept) are responsible. Beside the classical IgE-mediated anaphylactic shock there exists another anaphylactic shock, identical in its clinical picture and treatment but not in the mechanism of development. Epinephrine is the only effective drug in case of respiratory (bronchial asthma, laryngeal edema) or cardiovascular (hypotension, arrhythmias, hypovolemic shock) manifestation. It has to be administered as rapidly as possible. PMID- 7491456 TI - [Syncope: "missed death" or triviality?]. AB - Syncope is a common clinical problem defined as a transient loss of consciousness and postural tone with spontaneous recovery. Etiology is extremely diverse. In a great majority of cases the outcome is favorable; however, if a syncope is of cardiac origin, the risk of subsequent sudden death is relatively high. This fact implies an approach to the evaluation of the possible cause of syncope to minimize the risk of an important misdiagnosed problem. The etiology of syncope is generally found by careful history, clinical examination and an ECG. The strategy to use further investigations is discussed and provides a framework for approaching the final diagnosis prior to initiation of specific treatment. PMID- 7491462 TI - [Is beauty a measurable concept in medicine?]. PMID- 7491461 TI - [Nontraumatic coma in extramural practice]. AB - After a short review on pathophysiologic mechanisms of comatose states and their complications, a cohort of 392 comatose patients (Glasgow Coma Scale < or = 8) hospitalized in intensive care is analyzed in order to estimate the relative frequency of the different causes of nontraumatic coma. Depending on pathology, the following practical, sequential procedure is recommended: at first, identification and treatment of disorders of vital functions, objective estimate of the severity of the coma and rapid diagnostic orientation with a targeted neurologic investigation; then, simple therapeutic interventions in order to treat reversible causes of a metabolic encephalopathy as well as immediate measures for neuroprotection (anticonvulsive and antihypotensive therapy, oxygen, etc.). An initial, adequate control of the comatose patient is mandatory in order to limit disabling cerebral complications. PMID- 7491460 TI - [Rhythm emergencies]. AB - Arrhythmias have a high incidence in our population, and since the emergence of new methods in electrophysiology, the mechanism of most tachycardias can be identified. This allows not only to progress in the understanding of their pathophysiology but also in the treatment of arrhythmias, either in an emergency situation or in the chronic phase. The goal of this review is to give some therapeutic advice in case of rhythmic emergencies, keeping in mind that any arrhythmia should be subject to a cardiological work-up to seek specific treatment outside the emergency phase. The proposed therapies in this article are the ones which we are used to apply, but they differ depending on the areas. The final goal is to render service to the patients by trying to be as less damaging as possible, knowing that the understanding of the arrhythmogenic mechanism is not always possible. We hope that the little advice this review contains will be of use for the practitioner and for those who are confronted with patients suffering of acute arrhythmia. PMID- 7491463 TI - [Delirium: occurrence, diagnosis and therapy]. AB - Delirium, i.e. an acute confusional state, is frequently observed in a general hospital. Prevalence of delirium increases with higher age, usually in patients with cognitive deficits (e.g. signs indicative of some dementia) and particularly in those older patients with severe somatic illness. Symptoms of delirium are considerably uniform irrespective of different etiological causes; however, delirium is frequently underrecognized, which may lead to several complications and higher mortality rates. In this article the authors summarize data on prevalence, symptomatology as well as etiology and pathogenesis of delirium. In addition, syndromes which may have symptoms similar to delirium are discussed, and their treatment is recommended. PMID- 7491465 TI - [Cyclic sciatica and extrauterine endometriosis]. AB - Cyclic sciatica caused by ectopic endometriosis is an unusual cause of compression of a lumbar root, of the sciatic nerve or its plexus. A 38-year-old patient is presented with a history of menstruation associated with right-sided sciatica, which gradually developed over a 2 1/2-year period into disabling constant pain and a lumboradicular syndrome with signs of S1-root involvement. Suggested compression of the sciatic nerve by ectopic endometriosis in the sciatic notch region could be confirmed by CT and MR, by prompt response to hormonal suppression of ovarian function and by regression of the radiological findings. If the history of cyclic pain, local tenderness on deep palpation and the onset of referred pain in the buttock had been observed at the beginning, unnecessary and costly medico-technical procedures delaying diagnosis and treatment (including unnecessary physiotherapy) could have been avoided. Cyclic sciatica is discussed in a review of literature, and possibilities of treatment are discussed. PMID- 7491464 TI - [Study on the pretherapeutic differential diagnosis of adnexal tumors]. AB - How well do clinical examination, laboratory testing and imaging techniques predict the malignant or benign characteristics of an adnexal tumor before surgery? 60 patients underwent an extensive preoperative work-up including serum levels of gonadotropins, adrenal and ovarian steroids, oncofetal and placental antigens and CA 125. Sensitivity and specificity of laboratory criteria, including CA 125, compared unfavorably with others and are of no use in early diagnosis of ovarian malignancies. The differential diagnosis of adnexal tumors is primarily a matter of PV examination and ultrasound. PMID- 7491466 TI - [Treatment of arterial hypertension. Re-emergence of diuretics]. AB - Recent epidemiological studies on the management of arterial hypertension confirm the need to reserve special attention to diuretics in our prescription routine. The author analyzes briefly the eight essential points in favour of a first intention treatment of hypertension. The cost-efficiency ratio of diuretics remains extremely favourable, and the success in a reduction of arterial blood pressure is still comparable to that of other classes of antihypertensives. PMID- 7491467 TI - [Frequent airway infections]. AB - A 44-year-old patient experienced increasing shortness of breath and cough with yellow expectoration. Physical findings of the patient were not remarkable, whereas x-ray chest examination revealed cicatricial changes of the lower fields of the right lung. Laboratory findings showed a significant reduction of plasma gamma-globulin levels due to a global deficiency of all immunoglobulins. An infectious origin of the immunoglobulin deficiency was not detected. After exclusion of other acquired etiologic conditions, the diagnosis of a variable immunodeficiency syndrome was established. After antibiotic treatment with gyrase inhibitors, an immunoglobulin-substitution program was initiated. Immediately after the start of an immunoglobulin infusion, the patient developed an allergic reaction. Pretreatment with antihistamine drugs eliminated allergic symptoms. Following immunoglobulin treatment, incidence and severity of infectious diseases were significantly reduced. PMID- 7491469 TI - Complete denture occlusion: magic or planned? PMID- 7491470 TI - Fabrication of a full upper rehabilitation case with natural and implant abutments. PMID- 7491468 TI - [A case from practice (336)]. PMID- 7491471 TI - The North American Free Trade Agreement: will it affect the dental laboratory industry? PMID- 7491472 TI - Controlling workers compensation costs: a case study. PMID- 7491474 TI - Creating overdenture stability with a locking attachment. PMID- 7491473 TI - Anatomical and mechanical retention in maxillofacial prosthetics. PMID- 7491475 TI - Technique for fabricating a combination radiographic and surgical stent. PMID- 7491478 TI - Lyme disease research. PMID- 7491477 TI - A systematic approach to improved anterior esthetics. PMID- 7491476 TI - Don't fall into the overtime and penalty trap: contending with the salary vs. wage issue. PMID- 7491480 TI - Diet and test animals. PMID- 7491479 TI - Diet and test animals. PMID- 7491481 TI - Genetic discrimination: actuarial aspects. PMID- 7491482 TI - Scientific misconduct. Panel urges new approach to inquiries. PMID- 7491483 TI - Japan. Consortia to boost efforts to develop genome industry. PMID- 7491484 TI - Found: a star too small to shine. PMID- 7491485 TI - Why mammal ears went on the move. PMID- 7491487 TI - How does the Texaco case affect photocopying by scientists? The National Conference of Lawyers and Scientists. PMID- 7491486 TI - Brain researchers speak a common language. PMID- 7491489 TI - The interstellar carbon budget and the role of carbon in dust and large molecules. AB - Published data on stellar composition show that carbon in the sun is substantially more abundant than in other stars. A carbon abundance of 225 carbon atoms per 10(6) hydrogen atoms is representative of galactic stars, whereas published values for the sun range from 350 to 470 carbon atoms per 10(6) hydrogen atoms. Other elements are also present in enhanced quantities in the solar system, consistent with suggestions that a supernova event was closely associated with the formation of the solar system. The overabundance of carbon in the solar system has many important implications, including new constraints on nucleosynthesis models for supernovae and substantial modification of the so called "cosmic" composition normally adopted in discussions of galactic and interstellar abundances. A reduction in the galactic carbon budget, as suggested by the stellar composition data, strongly constrains the quantity of carbon that is available for the formation of interstellar dust, and some dust models now appear implausible because they require more carbon than is available. PMID- 7491488 TI - Molecular mimicry in protein synthesis? PMID- 7491490 TI - Venus reconsidered. AB - The Magellan imagery shows that Venus has a crater abundance equivalent to a surface age of 300 million to 500 million years and a crater distribution close to random. Hence, the tectonics of Venus must be quiescent compared to those of Earth in the last few 100 million years. The main debate is whether the decline in tectonic activity on Venus is closer to monotonic or episodic, with enhanced tectonism and volcanism yet to come. The former hypothesis implies that most radioactive heat sources have been differentiated upward; the latter, that they have remained at depth. The low level of activity in the last few 100 million years inferred from imagery favors the monotonic hypothesis; some chemical evidence, particularly the low abundance of radiogenic argon, favors the episodic. A problem for both hypotheses is the rapid decline of thermal and tectonic activity some 300 million to 500 million years ago. The nature of the convective instabilities that caused the decline, and their propagation, are unclear. PMID- 7491492 TI - Infrared spectrum of the cool brown dwarf Gl 229B. AB - Spectroscopic measurements of a cool brown dwarf, Gl 229B, reveal absorption features attributable to methane in the near infrared much like those of Jupiter. These features are not seen in any star. The presence of methane indicates that the surface temperature of Gl 229B is below 1000 kelvin. Features attributed to water vapor also indicate that Gl 229B is much cooler than any known star. PMID- 7491491 TI - Crystal structure of the ternary complex of Phe-tRNAPhe, EF-Tu, and a GTP analog. AB - The structure of the ternary complex consisting of yeast phenylalanyl-transfer RNA (Phe-tRNAPhe), Thermus aquaticus elongation factor Tu (EF-Tu), and the guanosine triphosphate (GTP) analog GDPNP was determined by x-ray crystallography at 2.7 angstrom resolution. The ternary complex participates in placing the amino acids in their correct order when messenger RNA is translated into a protein sequence on the ribosome. The EF-Tu-GDPNP component binds to one side of the acceptor helix of Phe-tRNAPhe involving all three domains of EF-Tu. Binding sites for the phenylalanylated CCA end and the phosphorylated 5' end are located at domain interfaces, whereas the T stem interacts with the surface of the beta barrel domain 3. The binding involves many conserved residues in EF-Tu. The overall shape of the ternary complex is similar to that of the translocation factor, EF-G-GDP, and this suggests a novel mechanism involving "molecular mimicry" in the translational apparatus. PMID- 7491493 TI - Autoencapsulation through intermolecular forces: a synthetic self-assembling spherical complex. AB - The synthesis and characterization of a system for the study of molecular recognition phenomena are described. The system involves a tetraurea molecule that is capable of assembly into various associated states through hydrogen bonding. In organic solvents, the dynamic transition between a low-ordered (aggregate) state and a highly ordered dimeric assembly can be induced by the introduction of smaller molecules of appropriate size and shape. These smaller molecules, such as benzene, adamantanes, and ferrocenes, act as guests that occupy the pseudospherical capsule formed by the dimeric host. Among various guests, those that best fill the cavity and offer chemical complementarity to the host are preferentially encapsulated. PMID- 7491494 TI - Yeast checkpoint genes in DNA damage processing: implications for repair and arrest. AB - Yeast checkpoint control genes were found to affect processing of DNA damage as well as cell cycle arrest. An assay that measures DNA damage processing in vivo showed that the checkpoint genes RAD17, RAD24, and MEC3 activated an exonuclease that degrades DNA. The degradation is probably a direct consequence of checkpoint protein function, because RAD17 encodes a putative 3'-5' DNA exonuclease. Another checkpoint gene, RAD9, had a different role: It inhibited the degradation by RAD17, RAD24, and MEC3. A model of how processing of DNA damage may be linked to both DNA repair and cell cycle arrest is proposed. PMID- 7491495 TI - Activation of the estrogen receptor through phosphorylation by mitogen-activated protein kinase. AB - The phosphorylation of the human estrogen receptor (ER) serine residue at position 118 is required for full activity of the ER activation function 1 (AF 1). This Ser118 is phosphorylated by mitogen-activated protein kinase (MAPK) in vitro and in cells treated with epidermal growth factor (EGF) and insulin-like growth factor (IGF) in vivo. Overexpression of MAPK kinase (MAPKK) or of the guanine nucleotide binding protein Ras, both of which activate MAPK, enhanced estrogen-induced and antiestrogen (tamoxifen)-induced transcriptional activity of wild-type ER, but not that of a mutant ER with an alanine in place of Ser118. Thus, the activity of the amino-terminal AF-1 of the ER is modulated by the phosphorylation of Ser118 through the Ras-MAPK cascade of the growth factor signaling pathways. PMID- 7491496 TI - Solution structure of the activator contact domain of the RNA polymerase alpha subunit. AB - The structure of the carboxyl-terminal domain of the Escherichia coli RNA polymerase alpha subunit (alpha CTD), which is regarded as the contact site for transcription activator proteins and for the promoter UP element, was determined by nuclear magnetic resonance spectroscopy. Its compact structure of four helices and two long arms enclosing its hydrophobic core shows a folding topology distinct from those of other DNA-binding proteins. The UP element binding site was found on the surface comprising helix 1, the amino-terminal end of helix 4, and the preceding loop. Mutation experiments indicated that the contact sites for transcription activator proteins are also on the same surface. PMID- 7491497 TI - A population genetic test of selection at the molecular level. AB - The role of natural selection in molecular evolution has been inferred primarily by rejection of null hypotheses based on neutral theory, rather than by acceptance of specific predictions based on selection. In this report, a population genetic test of a specific prediction for selection on DNA polymorphism is presented. Pyrethroid insecticide use constitutes an experiment for which form of selection and molecular target (voltage-gated sodium channels) are both known. As predicted, differential pyrethroid selection on tobacco budworm populations generated significant geographic heterogeneity in sodium channel marker allele frequencies, compared with arbitrary loci. PMID- 7491498 TI - Definition of two angiogenic pathways by distinct alpha v integrins. AB - Angiogenesis depends on cytokines and vascular cell adhesion events. Two cytokine dependent pathways of angiogenesis were shown to exist and were defined by their dependency on distinct vascular cell integrins. In vivo angiogenesis in corneal or chorioallantoic membrane models induced by basic fibroblast growth factor or by tumor necrosis factor-alpha depended on alpha v beta 3, whereas angiogenesis initiated by vascular endothelial growth factor, transforming growth factor alpha, or phorbol ester depended on alpha v beta 5. Antibody to each integrin selectively blocked one of these pathways, and a cyclic peptide antagonist of both integrins blocked angiogenesis stimulated by each cytokine tested. These pathways are further distinguished by their sensitivity to calphostin C, an inhibitor of protein kinase C that blocked angiogenesis potentiated by alpha v beta 5 but not by alpha v beta 3. PMID- 7491499 TI - Essential Ca(2+)-binding motif for Ca(2+)-sensitive inactivation of L-type Ca2+ channels. AB - Intracellular calcium (Ca2+) inhibits the opening of L-type (alpha 1C) Ca2+ channels, providing physiological control of Ca2+ entry into a wide variety of cells. A structural determinant of this Ca(2+)-sensitive inactivation was revealed by chimeric Ca2+ channels derived from parental alpha 1C and alpha 1E channels, the latter of which is a neuronal channel lacking Ca2+ inactivation. A consensus Ca(2+)-binding motif (an EF hand), located on the alpha 1C subunit, was required for Ca2+ inactivation. Donation of the alpha 1C EF-hand region to the alpha 1E channel conferred the Ca(2+)-inactivating phenotype. These results strongly suggest that Ca2+ binding to the alpha 1C subunit initiates Ca2+ inactivation. PMID- 7491501 TI - The evolution of molecular computation. PMID- 7491500 TI - Coactivator and promoter-selective properties of RNA polymerase I TAFs. AB - Human ribosomal RNA synthesis by RNA polymerase I requires the activator UBF and the promoter selectivity factor SL1, which consists of the TATA binding protein (TBP) and three associated subunits, TAFI110, TAFI63, and TAFI48. Here it is shown that both TAFI110 and TAFI63 contact the promoter, whereas TAFI48 serves as a target for interaction with UBF and is required to form a transcriptionally active SL1 complex responsive to UBF in vitro. TAFI48 also alters the ability of TBP to interact with TATA box elements, and the resulting complex fails to support transcription by RNA polymerase II. Thus, TAFI48 may function both as a target to mediate UBF activation and as a class-specific promoter selectivity factor. PMID- 7491502 TI - Hepatotoxicity of herbal remedies and mushrooms. PMID- 7491503 TI - Assessment of hepatic function with xenobiotics. PMID- 7491505 TI - Prospects of therapy of liver diseases with foreign genes. PMID- 7491504 TI - Targeting of drugs to the liver. PMID- 7491506 TI - Drug administration to liver patients: aspects of liver pathophysiology. PMID- 7491507 TI - Molecular basis of drug-induced immunological liver injury. PMID- 7491508 TI - Epidemiological identification and evaluation of hepatic adverse drug reactions. PMID- 7491509 TI - Mental illness and the continuum of residential stability. PMID- 7491510 TI - Social influences on bipolar affective disorders. AB - The impact of psychosocial adversity on the onset and course of bipolar disorder has been assessed in studies that have relied on methods of eliciting life event histories and evaluating family atmosphere. The results of life event studies have been inconsistent, perhaps because the relationship between bipolar disorder and major stress is only pronounced in first or early episodes. If this is so, this phenomenon itself invites explanation, whether in social or biological terms. The two studies to data of family atmosphere suggest an association between high expressed emotion and relapse. The relationship between psychosocial stress and bipolar disorder requires further and more detailed research. PMID- 7491511 TI - Increasing age is a risk factor for psychosis in the elderly. AB - We examined the association between ageing and the administrative incidence rate of late onset (after age 59) non-organic, non-affective psychosis in two samples of patients aged 60 years or older who were first admitted to hospital in (1) The Netherlands between 1978 and 1992 (n = 8010) and (2) nine regional health authorities in England and Wales (n = 1777) between 1976 and 1978. There was a linear trend in the association between increasing age and first admission rates for non-organic, non-affective psychosis in the elderly, after adjustment for the possible confounding effects of time trend and gender, corresponding to an 11% increase in the incidence with each 5-year increase in age. These observations support a connection between degenerative brain processes and onset of non affective psychosis in the elderly. PMID- 7491512 TI - Psychopathology and mortality in the general population. AB - Community studies of the relationship between mortality and mental disorders remain equivocal with mixed and unclear results. Longitudinal prospective data from the Epidemiologic Catchment Area Program are examined for the relationship between psychopathology and mortality. Analyses were performed with mortality at 1-year follow-up as the dependent variable using logistic regression. Specifically, the odds of dying from psychiatric disorders were considered while statistically adjusting for sociodemographic characteristics and treatment in primary or specialty medical care sectors. Significant predictors of dying were being male, older, and from low income households. Subjects with drug abuse or dependence were 12.4 times (CI = 2.9, 53.0) as likely to die as those without drug abuse/dependence. The odds of dying for those with major depressive disorder were over 2.6 times (CI = 1.1, 6.0) those without major depression. Separate analyses for all five communities and for only those over the age of 55 years showed similar results. PMID- 7491514 TI - Depression among Puerto Ricans in New York City: the Hispanic Health and Nutrition Examination Survey. AB - This study was conducted to analyze determinants of depression among Puerto Ricans by replicating and expanding earlier studies of depression among Cuban Americans and Mexican Americans. Data from the Hispanic Health and Nutrition Examination Survey, 1982-1984, were employed to examine depression and associated characteristics among Puerto Ricans. We utilized descriptive and multivariate statistics to examine the Center for Epidemiologic Studies Depression Scale (CES D)-assessed depressive symptomatology and the DSM-III/DIS specification of major depression. The findings indicated that CES-D-assessed depressive symptomatology among Puerto Ricans was associated with female gender, disrupted marital status, poor health, and lower socioeconomic status as indicated by low education, low household income, age, and unemployment. For both 6-month and 1-month DIS major depression, age, disrupted marital status, and income of less than $5,000 were significant risk factors. For 6-month DIS major depression, never married persons had a higher risk for depression. For 1-month diagnoses, writing Spanish better than English was associated with lower risk. In general, our findings for Puerto Ricans were similar to studies of depression among other Hispanic groups. We remained unable to explain the relatively extreme levels of depression among Puerto Ricans in New York, though several probable explanations are elaborated. We emphasized the general need to expand the range of research designs and current risk models in epidemiology in an effort to capture the complexity of psychosocial and cultural processes relevant to psychological distress. PMID- 7491515 TI - Assessing the need for frozen sections: still a valuable tool in thyroid surgery. AB - BACKGROUND: Since the development of fine-needle aspiration (FNA) there has been a trend away from frozen sections (FS) in the assessment of thyroid neoplasms. The objective of this study was to determine the role of FS in the surgical management of thyroid nodules in the presence of an adequate FNA biopsy finding. METHODS: Charts of patients who presented within a 3-year period for thyroid surgery were reviewed. Inclusion criteria consisted of both an adequate FNA and FS. RESULTS: Eighty-five patients met the inclusion criteria. Three lesions were benign, 71 were suspicious, and 11 were malignant with FNA. There were 66 deferred and 19 malignant diagnoses with FS. The overall accuracy for FNA and FS was 40% and 86%, respectively. When the FNA report was positive for malignancy, it was correct in 91% (10 of 11) of the cases. When the FNA report was suspicious, only 30% (21 of 71) had a malignant lesions. FS confirmed malignancy in 19 patients and deferred more extensive surgery in 66 patients with suspicious lesions. However, 18% of the deferred FS were found to be malignant on final pathology report. CONCLUSIONS: This study showed that there is a role for FS in the surgical management of thyroid nodules. Frozen sections can be useful when the FNA report is suspicious for malignancy; however, FS may be eliminated when the FNA report is positive for malignancy. PMID- 7491516 TI - Increased plasminogen activator and type IV collagenase activity in invasive follicular thyroid carcinoma cells. AB - BACKGROUND: An essential difference between benign and malignant follicular thyroid tumors is the ability to invade and metastasize. Thyrotropin (TSH) stimulates invasion of cultured human follicular thyroid cancer cells (FTC-133) via a protein kinase C (PKC) dependent mechanism. Tumor invasion depends on degradation of extracellular matrix by proteases. METHODS: We analyzed protease activity in FTC-133 and its more invasive clone, FTC-238. Cells were treated with TSH or 12-0-tetradecanoyl-phorbol-13-acetate (TPA), a PKC agonist, for 24 hours. Conditioned medium and cellular extract were subjected to substrate gel zymography with either casein-plasminogen or gelatin (collagen). Western blot and immunohistochemistry confirmed protease identity. RESULTS: We found increased 50 kd urokinase-like plasminogen activator (uPA) and 62 kd gelatinase activity by FTC-238 cells compared with the less invasive FTC-133 cells. There was no effect of TSH on uPA or collagenase activity at concentrations of 0.01 to 10 mU/ml. In both FTC-133 and FTC-238, TPA incubations of 0.1 to 100 ng/ml caused a dose dependent increase in uPA and a 94 kd type IV collagenase. CONCLUSIONS: These findings show that TSH-stimulated invasion may be due to PKC-induced activation of uPA and 94 kd type IV collagenase. uPA and basement membrane type IV collagenase warrant investigation as markers for follicular thyroid cancer. PMID- 7491513 TI - Cost-effectiveness over 10 years. A study of community-based social psychiatric care in the 1980s. AB - More and more, the chronically mentally ill are treated in community-based facilities. A review of international findings indicates that these treatments are on average considerably cheaper than traditional inpatient treatments. The present study adds some further evidence on the cost-effectiveness of a comprehensive programme of community-based treatments provided to a large number of mainly chronically mentally ill patients within a clearly defined catchment area over a 10-year period (1981-1990). The study revealed important needs, as mainly chronically ill, socially handicapped and frequently unoccupied patients, suffering from severe disorders were admitted to the different treatments. These needs were well compensated for by the offer of a differentiated network of crisis-intervention, half-way and community-based treatment facilities. Despite the low staff-patient ratios (0.04-0.5), a coherent pattern of transitions from admission to discharge indicated a good rehabilitative power of this network by diminishing referrals to hospitals and increasing referrals to outpatient and independent services. Performances and costs varied according to the structure of the different services. On average, direct daily costs for community-based social psychiatric care were about half the costs of inpatient treatment over the whole period. Whereas the mean costs for community treatment followed inflation rates, costs for inpatient treatment grew faster. Among the short-term effects, goals set at admission were reached to a great extent, while success rates for autonomy varied according to the different treatment philosophies. In the long-term, success rates for the different treatment facilities studied tended to be confirmed for independent living and work, but professional care was still needed by most of the patients. Despite certain methodological weaknesses of the present study, we suggest that funds for community-based social psychiatric treatment be maintained and we strongly recommend the further allocation of funds for this form of treatment. PMID- 7491517 TI - Molecular and cytogenetic characterization of a t(1;10;21) translocation in the human papillary thyroid cancer cell line TPC-1 expressing the ret/H4 chimeric transcript. AB - BACKGROUND: Activation of the ret proto-oncogene by three different chromosomal rearrangements occurs in up to 25% of papillary thyroid carcinomas. We developed a rapid screening technique to detect ret rearrangements in human interphase and metaphase cells on the basis of multicolor fluorescence in situ hybridization (FISH) of locus-specific DNA probes. METHODS: DNA from individual clones representing the respective ends of a yeast artificial chromosome (YAC) contig spanning the entire ret gene locus were labeled with either digoxigenin (visualized in red) or biotin (green) and hybridized to normal human lymphocytes and the papillary thyroid cancer cell line TPC-1 expressing the ret/H4 chimeric transcript. Further detailed analysis was performed with whole chromosome painting probes and locus-specific probes (YACs, P1s, DNA repeat probes) on tumor metaphase spreads. RESULTS: Hybridization of the YACs to unrearranged ret loci in normal human lymphocyte interphase nuclei showed two yellow domains because of probe overlap. Hybridization to TPC-1 interphase nuclei showed one yellow domain, and 1 red and 1 green domain separated by a large physical distance. Further analysis of metaphase spreads revealed a complex translocation t(1;10;21)(1pter > 1q31::21q22.1 > 21qter; 10q11.2 > 10pter::1q31 > 1qter; 21pter > 21q22.1;;10q21.2 > 10q11.2::10q21.2 > 10qter) and loss of the H4 gene locus on the nontranslocated chromosome 10. CONCLUSIONS: Break point spanning probes can reliably detect ret rearrangements in interphase nuclei. Locus-specific and whole chromosome painting probes can be used to further characterize complex rearrangements by fluorescence in situ hybridization to metaphase spreads. The papillary thyroid cancer cell line TPC-1 carries the paracentric inversion 10q, inv(10)(q11.2q21) and a complex t(1; 10; 21) translocation. Deletion of the H4 gene on the chromosome 10 not involved in the t(1; 10; 21) translocation suggests lack of normal H4 expression in the TPC-1 cell line. Further studies will have to address the role of the H4 gene product in tumor genesis and progression. PMID- 7491519 TI - Early treatment of hereditary medullary thyroid carcinoma after attribution of multiple endocrine neoplasia type 2 gene carrier status by screening for ret gene mutations. AB - BACKGROUND: Germline missense point mutations of the ret proto-oncogene have been shown as causative in multiple endocrine neoplasia type 2 (MEN 2A and 2B) and in familial medullary thyroid carcinoma (FMTC). Most of the mutations are found in exon 10, 11, or 16 of the gene and are easily recognized by restriction analysis. METHODS: Using restriction analysis, we screened 58 subjects from nine kindreds. RESULTS: Family members (n = 16) already known to be affected with the disease carried the germline mutation. Among the 42 subjects apparently unaffected, 37 were not gene carriers and 5 were gene carriers. Basal and pentagastrin stimulated serum calcitonin levels were normal in two patients and abnormal in three. All patients were treated with total thyroidectomy and central node dissection. In all cases multiple foci of MTC were shown at histologic examination. CONCLUSIONS: Our data indicate that genetic screening of MEN2 pedigrees allows the early identification of gene carriers. Because surgery of MTC in the preclinical phase has high probability of curing these patients, we suggest genetic screening soon after birth and total thyroidectomy in gene carriers as early as possible. PMID- 7491518 TI - Laparoscopic detection of hepatic metastases in patients with residual or recurrent medullary thyroid cancer. AB - BACKGROUND: After initial operations for medullary thyroid cancer (MTC), reoperation with removal of metastatic disease confined to the neck may benefit some patients. The identification of distant metastases precludes the possibility of curative reoperation. METHODS: Forty-one patients with hypercalcitoninemia after initial surgical treatment for MTC underwent laparoscopic (n = 36) or open (n = 5) examination and biopsy of the liver. Thirty-seven of these patients underwent imaging by computed tomography (CT), magnetic resonance imaging (MRI) of the liver, or both, and 17 underwent selective venous catheterization (SVC) with measurement of hepatic and peripheral vein stimulated calcitonin levels. RESULTS: Liver metastases were found in eight patients, seven by laparoscopy and one by open examination. Seven of these patients had normal CT or MRI scans of the liver. Laparoscopy or open liver examination revealed metastases in 2 of 11 patients with elevated hepatic vein-peripheral vein stimulated calcitonin ratios (greater than 1.3). Metastases appeared as small (less than 5 mm), bright white nodules on the surface of the liver. CONCLUSIONS: Direct examination and biopsy of the liver by laparoscopy may show small deposits of metastatic MTC in patients with normal CT and MRI scanning. PMID- 7491521 TI - DNA index and ploidy distinguish normal human parathyroids from parathyroid adenomas and primary hyperplastic parathyroids. AB - BACKGROUND: The goal of this study was to identify factors that might aid in diagnosis and intraoperative management of hyperparathyroidism. METHODS: We analyzed biopsy specimens of 242 parathyroids from 159 patients by use of flow cytometry and image cytometry (ICM) for DNA index (DI), defined as the content of nuclear DNA compared with that expected for a DNA diploid standard, for proliferative index (PI), and for ploidy (diploid versus aneuploid or tetraploid). RESULTS: True normal and normal parathyroids from patients with solitary adenomas were uniformly diploid. Abnormal ploidy (aneuploidy or tetraploidy) was identified frequently in adenomas and occasionally in hyperplasias with the exception that multiple endocrine neoplasia (MEN) biopsy specimens were uniformly diploid. DI for adenomas was similar to that for hyperplasias, and DI of both was higher than for normal glands. ICM-DI correlated positively with flow cytometry-DI and patient age and inversely with serum parathyroid hormone. PI was relatively low in all groups but was higher for hyperplasias versus normal parathyroids from patients with solitary adenomas and MEN versus non-MEN. PI correlated inversely with patient age. CONCLUSIONS: DI by ICM differentiates normal from abnormal parathyroids. DI might influence extent of resection in two- and three-gland hyperplasia and selection of the most appropriate gland for autografting and cryopreservation in patients with four gland hyperplasia. PMID- 7491522 TI - Outpatient thyroid and parathyroid surgery: a prospective study of feasibility, safety, and costs. AB - BACKGROUND: The purpose of this study was to determine feasibility, safety, and cost savings of outpatient thyroid and parathyroid surgery. METHODS: Consecutive unselected patients undergoing thyroid and parathyroid operations by two surgeons with a special interest in endocrine surgery were studied prospectively. RESULTS: One-hundred patients underwent operation, 61 as outpatients and 39 as inpatients. Outpatients included those undergoing thyroid lobectomy (39), total thyroidectomy (10), total thyroidectomy with parathyroidectomy (1), total thyroidectomy with modified neck dissection (1), and parathyroidectomy (10). Inpatients included those undergoing thyroid lobectomy (15), total thyroidectomy (8), total thyroidectomy with neck dissection (4), removal of substernal goiter (2), and parathyroidectomy (10). The average age of inpatients was slightly higher than that of outpatients (p < 0.05). Average hospital cost for outpatients was $1991 +/- $279 (range, $1594 to $2783) and for inpatients it was $2875 +/- 615 (range, $2031 to $4216), p < 0.001. Reasons for admission included extent of surgery (6), nausea (5), oversedation (4), urinary retention (2), inadequate home help (6), long travel time (2), patient preference (9), and medical reasons (5). No outpatients subsequently required admission. CONCLUSIONS: Outpatient thyroid and parathyroid surgery can be feasible and safe and resulted in a 30% savings in hospital costs. After extensive operations patients continue to require admission for postanesthetic complications, social reasons, or presence of serious comorbid disease. PMID- 7491520 TI - Cathepsin D in normal and neoplastic thyroid tissues. AB - BACKGROUND: Cathepsin D is a widely distributed lysosomal acidic endopeptidase. It is an estrogen-regulated protein that is a prognostic factor in breast cancer. The aim of this study was to measure cathepsin D concentrations in thyroid tissues and to correlate these concentrations with clinical and pathologic parameters. METHODS: Cathepsin D and thyroglobulin concentrations were measured in the cytosol of normal thyroid tissues (n = 14), benign nodules (n = 6), and thyroid carcinomas (n = 32) with an immunoradiometric assay. Statistical analysis was based on the Kruskal-Wallis and Wilcoxon tests and on the Spearman rank correlation coefficient. RESULTS: The mean level of cathepsin D, expressed as picomoles per milligram protein minus thyroglobulin, was higher in the 32 carcinomas, 29.1 +/- 15.5, than in the 14 normal thyroid tissues, 8.4 +/- 2.5 (p < 0.001) or in the 6 benign nodules, 11.2 +/- 7.3 (p = 0.003). Cathepsin D concentrations correlated with tumor size; Spearman rank correlation coefficient was rs = 0.44 (p = 0.012). No significant difference was found regarding histologic type. Cathepsin D concentrations were inversely correlated with the thyroglobulin level in the tumor; Spearman rank correlation coefficient was rs = 0.60 (p < 0.001). CONCLUSIONS: Cathepsin D concentration is higher in thyroid carcinoma than in normal thyroid tissue. Increased cathepsin D concentrations correlate with thyroid tumor size but not with histologic type. Further studies should be done to confirm the potential prognostic value of cathepsin D in patients with thyroid carcinomas. PMID- 7491523 TI - Expression and function of a CD3-like molecule on normal and abnormal human parathyroid cells. AB - BACKGROUND: Normal and abnormal human parathyroid tissue express the T-lymphocyte protein CD4, and parathyroid and lymphocyte cells show similarities with respect to mechanisms of calcium permeability and regulation of the cytoplasmic calcium concentration. METHODS: Anti-Leu4, a monoclonal antibody recognizing the T lymphocyte glycoprotein complex CD3, is used to immunohistochemically stain normal and abnormal human parathyroid cells and to explore influences on the parathyroid hormone (PTH) secretion of enzymatically dispersed parathyroid cells. RESULTS: Parathyroid glands of patients with different forms of hyperparathyroidism displayed variable expression of the anti-CD3 reactive complex. The stainings correlated both positively and inversely to immunoreactivity for a previously defined calcium sensor, the decreased expression of which may constitute a molecular basis for hyperparathyroidism. Incubation of parathyroid cells with the anti-Leu4 antibody inhibited PTH secretion and reduced its sensitivity to external calcium without influence on parathyroid cytoplasmic calcium concentration. CONCLUSIONS: The results suggest that the human parathyroid cells express a CD3-like molecule with the ability to interact in PTH release. PMID- 7491524 TI - Operative experience of U.S. general surgery residents in thyroid and parathyroid disease. AB - BACKGROUND: We wanted to determine whether the experience of general surgery residents is adequate and effective. METHODS: The Resident Statistic Summaries (Report C) of the Residency Review Committee (Surgery) for 8 academic years from 1986 through 1994 were analyzed. The main outcome measurements were total number of residents and programs, average number of operations performed, maximum number of operations performed, standard deviation, and the most common number of operations performed. RESULTS: For thyroidectomy the average per resident ranged from 10.3 to 12.6. The maximum ranged from 52 to 102. The standard deviations ranged from 6.96 to 8. The most common number of thyroidectomies performed ranged from 7 to 10 per graduating resident. For parathyroidectomy the average ranged from 4.1 to 5.1, the standard deviations were 3.44 to 4, the maximum ranged from 25 to 60, and the most common number performed was 2. CONCLUSIONS: U.S. graduates have highly variable experience in thyroid and parathyroid surgery. Most residents have inadequate experience in parathyroid surgery and marginal experience in thyroid surgery. PMID- 7491525 TI - Endoscopic retroperitoneal adrenalectomy. AB - BACKGROUND: The anterior transabdominal approach for adrenalectomy is associated with a longer postoperative recovery period than a posterior extraperitoneal adrenalectomy. The posterior approach is useful for patients requiring bilateral adrenalectomy or in those undergoing unilateral adrenalectomy for benign adenomas smaller than 5 cm. Recently transabdominal laparoscopic adrenalectomy has been used in patients with adrenal tumors. Endoscopic retroperitoneal adrenalectomy (ERA) is an alternative method that provides excellent exposure and should be associated with less postoperative morbidity. METHODS: Between 1993 and 1994 11 ERAs were performed in eight patients in the Department of Surgery, Istanbul Faculty of Medicine. The patients were placed in the prone semijackknife position on the operating table. After the retroperitoneal space was expanded with a balloon trochar, four 10 mm trochars were placed to perform the procedure. RESULTS: Among the eight female patients 23 to 65 years of age (mean, 42 +/- 12.4 years), three had bilateral adrenal hyperplasia caused by Cushing's disease, three patients had functioning adenoma, one patient had nonfunctioning adenoma (three on right and one on left adrenal), and one patient had right adrenal cyst. The mean operation time was 150 minutes (range, 90 to 300 minutes). No changes in PCO2 values have been found during intraoperative blood gas analyses. No intraoperative or postoperative complications occurred. All patients were discharged on the third postoperative day. CONCLUSIONS: ERA is a new and safe method of adrenalectomy. It is less invasive than the posterior approach. Patients treated by ERA seem to experience less postoperative pain and discomfort and have a shorter postoperative hospitalization and recovery period. PMID- 7491526 TI - Adrenal lesion in multiple endocrine neoplasia type 1. AB - BACKGROUND: Multiple endocrine neoplasia (MEN) type 1 is accompanied by adrenal involvement, but characteristics and clinical handling of this lesion have been insufficiently explored. METHODS: Patients with MEN 1 (n = 43) were monitored (mean, 6.3 years) with annual biochemical and radiologic adrenal evaluation. Adrenal specimens were examined by in situ RNA-RNA hybridization for expression of the MEN1 candidate gene phospholipase C beta 3 (PLC beta 3) and immunostaining for insulin-like growth factor-1 receptor. RESULTS: Altogether 17 patients (40%) displayed adrenal enlargement, which was limited to the adrenal cortex and showed signs of progression, marked atypia, and cancer development in three of them. Only the carcinoma exhibited adrenocortical hormone excess. PLC beta 3 was expressed in the hyperplastic and adenomatous proliferation but not the carcinoma. Pancreatic endocrine tumors with insulin-proinsulin excess were overrepresented in the patients with adrenocortical involvement, but significant insulin-like growth factor-1 receptor immunoreactivity was restricted to the carcinoma. CONCLUSIONS: The prevalent adrenocortical lesion associated with MEN 1 requires regular attention because of malignant potential. It was unrelated to loss of constitution heterozygosity for the MEN1 locus (11q13) and PLC beta 3 expression, except for the cortical carcinoma exhibiting allelic losses involving also the Wiedemann-Beckwith gene at 11p15. Mechanisms for mitogenic relationships between the pancreatic and adrenal lesions of MEN 1 demand further clarification. PMID- 7491527 TI - Surgical strategy for pheochromocytoma: emphasis on the pledge of flank extraperitoneal approach in selected patients. AB - BACKGROUND: Anterior transabdominal exploration has traditionally been advocated as the standard procedure for pheochromocytoma. However, some authors claim that a flank extraperitoneal approach with accurate unilateral localization is justifiable. METHODS: Retrospective analysis was performed on 87 patients with pheochromocytoma to determine the appropriateness of extraperitoneal exploration. RESULTS: There were 45 men and 42 women with a mean age of 44.7 years (range, 16 to 83 years). Fifteen patients (17.5%) had pheochromocytoma as a part of multiple endocrine neoplasia (MEN) type 2A, and four had familial pheochromocytoma, von Hippel-Lindau disease, or von Recklinghausen's disease. All tumors detected by preoperative localization studies were correctly identified and were resected through flank extraperitoneal (45 patients), transabdominal (28), thoracoabdominal (13), and posterior (1) approaches. Tumors were extraadrenal in 14, multiple in 22, bilateral in 12, and unequivocally malignant in 2 patients at the initial operation. Two patients died during the immediate postoperative period, giving an operative mortality of 2.3%. Two patients had persistent disease. During follow-up within a mean period of 5 years (range, 1 month to 13.7 years) 8 patients (9.1%) experienced recurrence or metastasis. Other than the patients with MEN 2, recurrence was not attributable to the operative approach. CONCLUSIONS: If a preoperative localization study is accurate, an extraperitoneal approach is justifiable for many patients with pheochromocytomas. PMID- 7491528 TI - Surgical management, DNA content, and patient survival in adrenal cortical carcinoma. AB - BACKGROUND: Surgical resection is the only potentially curative treatment for adrenal cortical carcinoma, yet the value of extended resection, palliative resection, and tumor DNA analysis remains unclear. METHODS: The records of 23 patients with adrenal cortical carcinoma who underwent primary surgical resection at our institution were retrospectively reviewed. Flow cytometric DNA analysis was performed on primary tumor tissue from 14 patients. RESULTS: Sixteen of 23 patients underwent complete resection. For these 16 patients the median follow-up was 43 months, the actuarial median survival was 46 months, and the actuarial 5 year survival rate was 46%. The seven patients who underwent incomplete resection all died of disease with a median survival of 8.5 months. Isolated local recurrence as the first site of failure occurred in two patients. Only completeness of resection (p = 0.004) and stage at presentation (p = 0.006) were significant prognostic indicators. None of the following predicted a poor prognosis in patients who underwent complete resection: (1) need for extended resection, (2) presence of renal vein or inferior vena cava tumor thrombus, or (3) tumor aneuploidy (14 of 14 tumors were aneuploid). CONCLUSIONS: Long-term survival is possible in patients with adrenal cortical carcinoma if complete, margin-negative tumor resection can be achieved. Isolated local recurrence is uncommon after complete resection. Because adrenal cortical carcinomas are consistently aneuploid, tumor DNA content is not a useful prognostic factor. PMID- 7491529 TI - Presymptomatic DNA screening in families with multiple endocrine neoplasia type 2 and familial medullary thyroid carcinoma. AB - BACKGROUND: Missense mutations of the ret proto-oncogene on chromosome 10q11.2 are the underlying cause of hereditary medullary thyroid carcinoma (MTC), either as familial MTC only (FMTC) or as a part of multiple endocrine neoplasia type 2 syndrome (MEN 2). This study presents our experience with direct presymptomatic DNA screening in MEN 2 and FMTC kindreds. METHODS: Twenty one families with MEN 2 or FMTC were considered in the study. One hundred three individuals had been analyzed; 56 were at risk. The ret mutations were detected by DNA analysis of exons 10, 11, and 16 by using nonradioactive labeling method based on digoxigenin DNA sequencing technique. Serum calcitonin evaluation was carried out in all individuals at risk. Thyroidectomy was performed in those who had to undergo surgery. RESULTS: The ret mutations were identified in all 21 families. In MEN 2A and FMTC families mutations occurred in exons 10 and 11. MEN 2B families had mutations in exon 16. The most frequent mutation in MEN 2A and FMTC affected codon 634. Twenty one gene carriers were identified in unaffected individuals at risk. Ten of 21 gene carriers had elevated calcitonin levels, and 11 had normal levels. MTC or C-cell hyperplasia was found in six gene carriers with pathologic calcitonin values who underwent operation. In a 5-year-old gene carrier with normal calcitonin values C-cell hyperplasia was evident. CONCLUSIONS: Direct predictive DNA analysis allows us to identify MEN 2 or FMTC gene carriers and offer them prophylactic treatment. PMID- 7491530 TI - Indium-111-pentetreotide scanning versus conventional imaging techniques for the localization of gastrinoma. AB - BACKGROUND: The present study evaluates 111In-pentetreotide scanning as a method for detection of gastrinomas. Operative findings serve as the benchmark for comparison of the efficacy of 111In-pentetreotide versus conventional imaging studies. METHODS: Twelve patients (seven female and five male; age, 37 to 80 years) with histologic confirmation of gastrinoma underwent thin section dynamic computed tomography (CT) scanning and 111In-pentetreotide scanning. The sensitivity, specificity, positive predictive value, negative predictive value, and accuracy of 111In-pentetreotide and CT scanning are compared on the basis of tumor size and location. RESULTS: Thirty discrete foci of intrahepatic and extrahepatic tumors were detected at operation. CT scanning detected three of nine pancreaticoduodenal lesions, whereas eight of these nine extrahepatic primary tumors were imaged by 111In-pentetreotide scanning. No false-positive 111In-pentetreotide scans were noted. The sensitivity of CT scanning for detection of metastatic disease was 56% versus 94% for the 111In-pentetreotide scan. Successful CT imaging was highly dependent on tumor size. No tumor smaller than 1 cm was imaged by CT, whereas four of seven lesions greater than 1 cm were imaged by 111In-pentetreotide scintigraphy. The smallest gastrinoma imaged by 111In-pentetreotide scanning was a 4 mm duodenal tumor. CONCLUSIONS: 111In pentetreotide scanning was superior to CT scanning for localizing gastrinomas. Further studies are required to determine whether 111In-pentetreotide scans will complement or replace traditional imaging methods. PMID- 7491531 TI - Persistent or recurrent hyperparathyroidism in patients with double adenomas. AB - BACKGROUND: Are double parathyroid adenomas a discrete clinical entity or are all hyperplastic parathyroid glands of varying size? This investigation was done to determine whether patients with persistent or recurrent hyperparathyroidism and double adenomas (DA) differ in clinical profile or in response to treatment from patients with hyperplasia. METHODS: From 1982 to 1993, 37 unselected patients with persistent (11) or recurrent hyperparathyroidism (26) were treated. Twenty one had DA and 16 had hyperplasia. DAs were defined as two abnormal and two normal parathyroid glands. RESULTS: Patients with persistent or recurrent hyperparathyroidism caused by DA were older and had more muscle weakness, neuropsychiatric disorders, constipation, and weight loss (p < 0.001) than patients with persistent or recurrent hyperparathyroidism caused by hyperplasia. Nephrolithiasis was more common in patients with recurrent hyperparathyroidism caused by hyperplasia than in patients with recurrent hyperparathyroidism caused by DA (p < 0.001). Serum Ca2+ levels before operation were similar in these groups with variable parathyroid hormone levels. No recurrences occurred in either group (DA [mean, 6 yr]; hyperplasia [mean, 5 yr]). Postoperative levels of serum Ca2+ and parathyroid hormone were normal in both groups. CONCLUSIONS: Patients with persistent or recurrent hyperparathyroidism caused by DA are older, have different clinical manifestations, and are cured by resection of the second adenoma. These findings support the concept that DA and hyperplasia are distinct entities. PMID- 7491533 TI - Prognostic factors and risk group analysis in follicular carcinoma of the thyroid. AB - BACKGROUND: The understanding of prognostic factors has facilitated stratification of risk groups in differentiated carcinoma of the thyroid. The prognostic factors have clearly identified the risk groups as low, intermediate, and high risk. Risk group categorization has facilitated a selective surgical approach for thyroid carcinoma. METHODS: A retrospective review of 228 patients with follicular carcinoma of the thyroid was undertaken. Various prognostic factors and risk groups were analyzed. Univariate and multivariate analyses were performed, and the survival curves were plotted by the Kaplan-Meier method. Fifty nine (26%) patients presented with Hurthle cell histology. The risk groups revealed 62 patients in the low, 84 in the intermediate, and 82 in the high risk groups. RESULTS: The 10-year survival for low, intermediate, and high risk groups was 98%, 88%, and 56%, respectively, and the 20-year survival for the same groups was 97%, 87%, and 49%, respectively. Adverse prognostic factors included age older than 45 years (p < 0.001), Hurthle cell variety (p = 0.05), extrathyroidal extension, tumor size exceeding 4 cm, and the presence or absence of distant metastasis (p < 0.001). Gender, focality, and presence of lymph node metastasis had no significant impact on prognosis. CONCLUSIONS: Patients in the low risk group have excellent survival, whereas the high risk group behaves poorly. Appropriate selection of treatment for the primary disease and adjuvant therapy should be considered on the basis of the prognostic factors and risk group analysis. PMID- 7491532 TI - Role of surgery in management of adrenocorticotropic hormone-producing islet cell tumors of the pancreas. AB - BACKGROUND: Ectopic adrenocorticotropic hormone-producing islet cell tumors of the pancreas (ACTH-ICT) are a rare cause of Cushing's syndrome with a severe and rapidly progressive clinical course. METHODS: Charts were reviewed on all patients evaluated and treated for proven Cushing's syndrome caused by ACTH-ICT (n = 12), specifically for the role of surgery in the management of this disease. RESULTS: Ten (83%) of twelve patients with ACTH-ICT had liver metastases at the time of diagnosis (eight of eight with Zollinger-Ellison syndrome, two of four without Zollinger-Ellison syndrome). Surgical management of the primary tumor included three patients who underwent distal pancreatectomy combined with hepatic resection and one patient who underwent laparoscopic enucleation of a tumor from the pancreatic tail. Eight of twelve patients underwent bilateral adrenalectomy to control symptoms of Cushing's syndrome, including three patients who underwent concurrent distal pancreatectomy and hepatic resection. Six of twelve patients died of the disease within 2 1/2 years of diagnosis, four are alive with progressive hepatic metastases, and one has biochemical evidence of disease. CONCLUSIONS: ACTH-ICT of the pancreas is an aggressive tumor, particularly when there is coproduction of gastrin. The benefit of aggressive surgical resection of primary or metastatic ACTH-ICT has not been established. However, palliative bilateral adrenalectomy is justified, because no patients had biochemical cures after aggressive surgical resection in this series. PMID- 7491536 TI - Correct preoperative localization: does it permit a change in operative strategy for primary hyperparathyroidism? AB - BACKGROUND: A meticulous bilateral neck exploration by an experienced endocrine surgeon offers a high cure rate with low morbidity for patients with primary hyperparathyroidism. The advent of localizing studies raises the possibility of unilateral neck exploration. The cost-effectiveness of preoperative localizing studies and unilateral neck exploration in primary hyperparathyroidism are controversial issues. This study was designed to determine the risks of missing a contralateral pathologic parathyroid gland in patients with preoperative localization that was confirmed at neck exploration. METHODS: Preoperative studies (ultrasonography, nuclear radioactive imaging scan, or both) were performed in 79 patients with primary hyperparathyroidism. In 58 patients a definite localization of an enlarged parathyroid gland was confirmed at operation. All patients underwent a meticulous bilateral neck exploration. RESULTS: Unilateral neck exploration was feasible only in 73.4% of the patients, according to our localizing modalities, and an additional enlarged parathyroid gland on the contralateral side, not detected before operation, was revealed in five patients (8.6%). False-positive rates were 1.7% for ultrasonography and 13% for scan. CONCLUSIONS: These results indicate an unacceptably high surgical failure rate for unilateral neck exploration guided by preoperative localizing studies compared with a bilateral neck exploration by an experienced endocrine surgeon, questioning the cost-effectiveness of preoperative localizing studies. PMID- 7491534 TI - Normocalcemic hyperparathyroidism. PMID- 7491535 TI - Hyperparathyroidism of multiple endocrine neoplasia type 1: candidate gene and parathyroid calcium sensing protein expression. AB - BACKGROUND: Hyperparathyroidism affects most patients with multiple endocrine neoplasia type 1 (MEN 1). This study investigates expression of the candidate MEN1 gene phospholipase C beta 3 (PLC beta 3) and expression and function of a putative calcium sensing protein (CAS) in hyperparathyroidism of MEN 1. METHODS: In 31 parathyroid glands from 17 patients with MEN 1, CAS distribution was studied immunohistochemically and parallel sections were explored for PLC beta 3 mRNA expression by in situ hybridization. Enzymatically dispersed parathyroid cells were analyzed for cytoplasmic calcium concentrations [Ca2+]i and parathyroid hormone (PTH) release. RESULTS: All glands exhibited a heterogeneously reduced CAS immunoreactivity, especially meager in nodularly assembled parathyroid cells. Calcium regulated [Ca2+]i and PTH release tended to be more deranged in the glands possessing the lowest immunostaining. Parathyroid PLC beta 3 invariably was homogeneously expressed, and this included even MEN 1 patients with reduced PLC beta 3 expression in endocrine pancreatic tumors. CONCLUSIONS: The findings support variable calcium insensitivity of [Ca2+]i and PTH release in hyperparathyroidism of MEN 1, apparently coupled to heterogeneously reduced CAS expression. For clarification of the role of PLC beta 3 in MEN 1 parathyroid tumorigenesis further study of this protein is required. PMID- 7491537 TI - Multiple endocrine neoplasia type 2B: more than an endocrine disorder. AB - BACKGROUND: Multiple endocrine neoplasia type 2B (MEN 2B) is a rare disorder differentiated from MEN 2A primarily by its extraendocrine features. This report describes the clinical spectrum and outcome of MEN 2B. METHODS: Twenty-one patients underwent operation for manifestations of MEN 2B between 1970 and 1993. Median follow-up was 16.9 years. Diagnosis was made through family screening in nine, the development of medullary thyroid carcinoma (MTC) in seven, phenotypic features in four, and constipation in one. Median age at presentation of colonic dysfunction, MTC, and pheochromocytoma was 0.1, 16, and 28 years, respectively. RESULTS: Every patient had MTC. Fifteen (94%) of 16 patients undergoing primary thyroidectomies had multicentric disease, and seven (44%) had nodal metastases. Seven patients (33%) had pheochromocytoma, six bilateral and one malignant. Adrenalectomy was curative in every patient. Nineteen patients (90%) had colonic disturbances, typically chronic constipation from birth. Megacolon developed in 14 patients, and eight required colonic surgery. Every patient had the characteristic phenotype. Dominant features included neuromas of the tongue, buccal mucosa, lips, conjunctivae, and eyelids and a marfanoid habitus. Other features included high arched palate, corneal nerve thickening, and dental and skeletal abnormalities. Four patients died, two of metastatic MTC, one after operation for metastatic MTC, and one as a consequence of colonic perforation. Of 17 survivors, three have hepatic metastases from MTC, eight have nodal metastases, and six are well with normal or mildly elevated calcitonin levels. CONCLUSIONS: MEN 2B is characterized by a relatively aggressive form of MTC, bilateral pheochromocytoma, severe colonic dysfunction, and a multitude of other extraendocrine abnormalities. Early recognition of MEN 2B and early prophylactic thyroidectomy are essential. Colonic dysfunction has previously received little attention, and further investigation of the pathogenesis and treatment of this disorder is warranted. PMID- 7491538 TI - Evaluation of total/near-total thyroidectomy in a short-stay hospitalization: safe and cost-effective. AB - BACKGROUND: Once the decision of perform total/near-total thyroidectomy has been made, common perioperative management strategies include frequent postoperative laboratory determinations, bedside airway adjuncts, and hospital stays of about 3 days. We propose a regimen for safe, cost-effective, short-stay total/near-total thyroidectomy. METHODS: One hundred fifty total/near-total thyroidectomies performed between 1991 and 1994 were studied to test our short-stay thyroidectomy regimen. Patients were admitted the day of operation and observed overnight. Serum calcium values were obtained at 8, 14, and 20 hours after operation. Twenty three-hour discharge criteria included no wound or airway problems, stable vital signs, tolerance of normal diet and activity, and an upsloping serum calcium curve. RESULTS: Of 150 patients undergoing total/near-total thyroidectomy, 145 (97%) met 23-hour discharge criteria. No deaths (0%) occurred. Overall morbidity (six patients [4%)]) included one (0.7%) patient with postoperative hemorrhage, one (0.7%) patient with recurrent laryngeal nerve injury, three (2%) patients with transient hypocalcemia, and one (0.7%) patient with permanent hypocalcemia. Average length of stay was 1.06 days. CONCLUSIONS: Significant airway and wound problems rarely develop beyond the first 12 to 18 hours after total/near-total thyroidectomy. Serial serum calcium determinations used to construct a three point calcium curve at 20 hours after operation can reliably and safely identify patients at risk to have clinically significant hypocalcemia. Total/near-total thyroidectomy can be performed safely in a short-stay, 23-hour hospitalization setting with substantial cost savings. PMID- 7491539 TI - Undescended superior parathyroid gland: a potential cause of failed cervical exploration for hyperparathyroidism. AB - BACKGROUND: The undescended inferior parathyroid gland is a well-established although infrequent embryologic abnormality resulting in an ectopically located gland usually associated with thymic tissue and is found most commonly within the carotid sheath at the level of the carotid bifurcation. Embryologically undescended superior glands (either normal or enlarged), clearly above the level of the upper pole of the thyroid gland and found within or in approximation to the lateral pharyngeal wall, have not been previously described. METHODS: The locations of all parathyroid glands at the time of cervical exploration were carefully examined as to embryologic origin in a combined experience of more than 3000 patients with primary or secondary hyperparathyroidism. RESULTS: Five patients had enlarged abnormal glands located within or in close approximation to the pharyngeal wall. All were above the upper pole of the thyroid gland. Two were identified at reoperation on the basis of localization studies. One gland was found after resection of a massive concomitant cervical goiter. The other two were found after an extensive search for a superior gland. CONCLUSIONS: Undescended superior glands are extremely rare (0.08%). They are located within or in approximation to the pharynx above the level of the thyroid gland with an unusual blood supply, which supports an embryologic rather than acquired cause for their ectopic location. PMID- 7491540 TI - An immunohistochemical survey for neuroendocrine cells in regional pancreatic lymph nodes: a plausible explanation for primary nodal gastrinomas? Mayo Clinic Pancreatic Surgery Group. AB - BACKGROUND: The existence of primary gastrinomas in lymph nodes continues to be controversial, because distinction from metastases from occult or regressed primary tumors may be difficult to exclude with certainty. If primary nodal gastrinomas do indeed occur, precursor neuroendocrine cells should be identifiable within lymph nodes. In a effort to detect these cells we undertook an immunohistochemical survey of regional pancreatic lymph nodes by using antibody against chromogranin, a highly specific marker for mature neuroendocrine cells. METHODS: We retrospectively identified consecutive cases from five surgeons in which Whipple resections had been performed for nonendocrine pathologic conditions. All formalin-fixed, paraffin-embedded lymph nodes were identified from these 106 cases, excluding lymph nodes with metastases. Immunoperoxidase staining with monoclonal antibody against chromogranin A was performed on single tissue sections. All slides were reviewed by a single pathologist; chromogranin-positive cells were identified and, when possible, further evaluated immunohistochemically with additional neuroendocrine markers, including sequential synaptophysin staining. RESULTS: A total of 1026 lymph nodes were available for review. Although well-formed intranodal chromogranin-positive nests were not identified, six nodes from three patients contained 13 strongly chromogranin-positive cells representing putative neuroendocrine cells. Mast cells showed weak nonspecific staining with chromogranin but were morphologically distinct. Perinodal adipose tissue contained a single paraganglion and several small neuroendocrine clusters. CONCLUSIONS: Putative neuroendocrine cells are rarely (less than 1%) found in regional pancreatic lymph nodes. Although these may be embryonic rests and might represent precursors of primary nodal gastrinomas, specific hormones have yet to be identified in these cells. Although these data provide support for the concept of primary nodal gastrinomas, this remains a diagnosis of exclusion. PMID- 7491541 TI - Clinical islet transplantation experience of the University of California Islet Transplant Consortium. AB - BACKGROUND: The University of California Islet Transplant Consortium was formed to evaluate the feasibility of performing clinical islet transplantation at different transplant centers by using a single centralized islet isolation laboratory. METHODS: From July 1992 through February 1995 seven adult islet transplantations were performed, six allografts and one autograft. Once procured, human pancreata were brought to the UCLA-VA Islet Core Laboratory for islet isolation and purification, which were then transported to different centers for transplantation. Patients 1 through 3 received their transplants in Los Angeles, patient 4 received her islet transplant in Torrance, and patients 5 through 7 received their transplants in San Francisco. RESULTS: Although none of these patients achieved insulin independence, four of seven had functioning grafts longer than 6 months as indicated by circulating C-peptide level greater than 0.7 ng/ml. Furthermore, improved glucose control as shown by a decreased insulin requirement was seen in 57% (four of seven patients) of these patients. The ability to isolate islets at a single laboratory and transport them long distances to different centers was shown in patients 4 through 7. CONCLUSIONS: Islet transplantation can be performed with improvements in blood glucose control, and islets can be isolated at a centralized location and successfully transported to different centers for transplantation. PMID- 7491542 TI - Prospective study of surgical resection of duodenal and pancreatic gastrinomas in multiple endocrine neoplasia type 1. AB - BACKGROUND: The role of surgical resection of gastrinoma in multiple endocrine neoplasia type 1 (MEN 1) is controversial because of low biochemical cure rates, but with adequate duodenal exploration higher cure rates may be possible. METHODS: We have prospectively evaluated this proposal in ten consecutive patients with MEN 1 and Zollinger-Ellison syndrome who underwent surgical exploration for gastrinoma resection including a detailed evaluation of the duodenum by palpation, intraoperative endoscopy with transillumination, and duodenotomy. RESULTS: Duodenal tumors were present in seven patients. Six of seven patients had metastatic deposits in lymph nodes, and two of seven had synchronous pancreatic tumors. Three patients had a single duodenal tumor, one patient had two tumors, and three patients had more than 20 duodenal tumors. Positive gastrin staining by use of immunohistochemistry was seen in all duodenal tumors. None of these seven patients were biochemically cured. Of three patients with negative duodenal explorations, two had single pancreatic tumors removed and one had only lymph node gastrinoma. No patients were biochemically cured. CONCLUSIONS: Not all patients with MEN 1 and Zollinger-Ellison syndrome have duodenal gastrinomas. In the 70% of patients with duodenal tumors, even extensive duodenal exploration with removal of positive lymph nodes does not result in cures because 86% of tumors had metastasized to lymph nodes and 43% of patients had large numbers of tumors. PMID- 7491543 TI - Role of preoperative iodocholesterol scintiscanning in patients undergoing adrenalectomy for Cushing's syndrome. AB - BACKGROUND: Iodocholesterol scintiscanning (IS) is a noninvasive, functional diagnostic test. We report our experience with IS as an adjunct for adrenal surgery for Cushing's syndrome. METHODS: Between April 1983 and October 1994, 23 patients with Cushing's syndrome from benign primary adrenal disease underwent IS and computed tomography (CT) and/or magnetic resonance imaging (MRI). Twelve patients had unilateral adrenal involvement with a solitary adenoma (n = 11) or unilateral multinodular disease (n = 1), and 11 patients had bilateral adrenal disease. RESULTS: In patients with Cushing's syndrome caused by unilateral adrenal disease, IS was 100% specific and 100% sensitive, whereas in two cases CT/MRI incorrectly showed bilateral disease. In patients with Cushing's syndrome with bilateral adrenal involvement, IS had one false-negative result with nonvisualization. CT/MRI showed unilateral disease in four cases and no abnormalities in two. All patients in this series were cured of Cushing's syndrome after unilateral adrenalectomy in 11 cases and bilateral adrenalectomy in 12 cases. CONCLUSIONS: IS is a highly sensitive and specific imaging modality and is an essential adjunct to biochemical testing in planning adrenal resections for Cushing's syndrome caused by primary adrenal disease. PMID- 7491548 TI - Cystic fibrosis: transition from paediatric to adult physician's care. PMID- 7491544 TI - Adrenocorticotropic hormone-secreting pheochromocytomas: the exception to the rule. AB - BACKGROUND: Operative management of pheochromocytomas dictates resection of the involved adrenal and exploration-resection of the contralateral gland if enlarged. We describe an exception to this rule. METHODS: We report the largest series of patients with adrenocorticotropic hormone (ACTH)-secreting pheochromocytomas and review the world literature. RESULTS: Four patients presented with findings of adrenocorticoid and catecholamine excess, as well as elevated levels of plasma ACTH, urinary metanephrines, and urinary free cortisol. Abdominal computed tomography scans revealed bilateral adrenal hyperplasia, and magnetic resonance imaging scans showed a unilateral adrenal mass with a bright T2 signal suggesting a pheochromocytoma. Two patients underwent adrenal venous sampling localizing ACTH secretion to the pheochromocytoma. All underwent unilateral adrenalectomy for a benign tumor without morbidity or death, leaving the contralateral hyperplastic adrenal in situ. After operation all patients experienced normalization of their levels of plasma ACTH, urinary metanephrines, and urinary free cortisol with resolution of symptoms. Combining our series with previously reported cases of ACTH-secreting pheochromocytomas, almost all are benign (20 of 21), in contrast to most ACTH-secreting tumors. CONCLUSIONS: ACTH secreting pheochromocytomas are the exception to the rule; unilateral adrenalectomy is curative and the contralateral hyperplastic adrenal can be preserved. This approach results in resolution of both syndromes of hormone excess and preserves long-term adrenal function. PMID- 7491545 TI - Large cystic/solid thyroid nodules: a potential false-negative fine-needle aspiration. AB - BACKGROUND: False-negative fine-needle aspiration (FNA) biopsy results of thyroid nodules are of particular concern because they imply missed malignant lesions. The purpose of this study was to identify characteristics of thyroid nodules that may lead to false-negative cytologic interpretation after FNA. METHODS: We reviewed 90 consecutive patients who underwent preoperative FNA of thyroid nodules followed by thyroid surgery during a period of 27 months, including their clinical data, type and size of thyroid nodule, FNA cytology results, and final pathology report of the surgical specimen. RESULTS: Thyroid nodules that had the highest probability of malignancy were those that were large (3 cm or larger), cystic/solid, or large and cystic/solid. The overall false-negative rate for preoperative FNA was 11%. Large, cystic/solid, and thyroid nodules with both characteristics had false-negative rates of 17%, 25%, and 30%, respectively, compared with 0%, 9%, and 17% for small (less than 3 cm), solid, and solid nodules 3 cm or larger, respectively. CONCLUSIONS: Because of the high prevalence of malignancy in thyroid nodules that are large (3 cm or larger), cystic/solid, or large and cystic/solid and the high false-negative rate of FNA in diagnosing these lesions, thyroid lobectomy for diagnosis should be strongly considered in these patients even when FNA cytologic finding is interpreted as benign. PMID- 7491546 TI - Inflammatory predictors for the development of the adult respiratory distress syndrome. PMID- 7491547 TI - Childhood asthma: what is it and where is it going? PMID- 7491549 TI - Cytokine expression in normal, atopic, and asthmatic subjects using the combination of sputum induction and the polymerase chain reaction. AB - BACKGROUND: The importance of cytokines in the asthmatic inflammatory response is becoming apparent. The aim of this study was to determine whether the non invasive method of induced sputum combined with the polymerase chain reaction would allow the detection of messenger RNA (mRNA) encoding a range of cytokines on a qualitative basis. METHODS: Four groups were studied comprising 10 normal subjects, six atopic, 10 mild and five moderately severe asthmatic subjects. Sputum was induced by the inhalation of nebulised 3.5% saline and total RNA extracted from the expectorated cells. Expression of cytokine message within induced sputum was examined by reverse transcription and the polymerase chain reaction (PCR) using primers specific for a range of cytokines (IL-1 alpha, IL-2, IL-3, IL-4, IL-5, IL-6, IL-8, RANTES, TNF alpha, IFN alpha, IFN gamma). Presence or absence of the signal was determined at 35 and 70 cycles of PCR by electrophoretic size fractionation on ethidium bromide stained agarose gels. RESULTS: Cytokine message was detectable in sputum by this method. All samples showed a positive result for actin control. Analysis of signal for the cytokines in all subjects showed that, at 70 cycles, IL-1, IL-5, IL-8, and TNF alpha were detected in more subjects than would be expected by chance. IL-5 mRNA was detected in more of the asthmatic patients (moderate 80%, mild 40%) than in the atopic subjects (33%), who in turn showed expression of this cytokine in more individuals than nonatopic subjects (10%). CONCLUSIONS: The combination of sputum induction and PCR appears to be a useful, non-invasive tool to explore the chronic inflammation of asthma and possibly other lung disorders. It should enable differences between normal and asthmatic subjects to be identified for future confirmation by quantitative techniques. PMID- 7491550 TI - The tar reduction study: randomised trial of the effect of cigarette tar yield reduction on compensatory smoking. AB - BACKGROUND: Observational and short term intervention studies have reported that smokers of low tar cigarettes inhale more deeply (that is, compensate) than those who smoke high tar cigarettes. To quantify this effect a long term randomised trial was conducted on the effects of switching to low tar cigarettes. METHODS: The trial was carried out between April 1985 and March 1988 among cigarette smokers in the British Civil Service, measuring blood carboxyhaemoglobin (COHb) levels and serum cotinine levels as markers of tobacco smoke intake. Volunteers first switched to a cigarette brand yielding around 10% less tar than their usual brand to identify smokers able to change brand. The 434 subjects who successfully switched were then randomly allocated to one of three groups: (a) "fast reduction" group which changed to a brand of cigarettes with a tar yield of about half that of their usual brand; (b) "slow reduction" group which reduced to the same level in steps over several months; and (c) a control group which continued smoking cigarettes with a tar yield 10% lower than their usual brand. RESULTS: Over the course of the trial cigarette consumption declined slightly in all three groups. In both the "fast reduction" and the "slow reduction" groups, intake of COHb and cotinine was reduced, though not to the same extent as the yield reduction. Comparison of the results before randomisation with those at the end of the trial showed that a reduction in carbon monoxide yield of 45% was associated with a decrease in carbon monoxide intake of 19% (95% confidence interval 14% to 24%) and that a reduction in nicotine yield of 40% was associated with an 11% (6% to 16%) reduction in nicotine intake, reflecting relative intakes of about 1.5 for both carbon monoxide and nicotine in the "fast reduction" group. Results were similar in the "slow reduction" group with a 42% reduction in carbon monoxide yield, a 16% (11% to 22%) reduction in carbon monoxide intake, a 37% reduction in nicotine yield, and a 6% (0% to 13%) reduction in nicotine intake. Estimates of compensation derived from these results were 65% for carbon monoxide, 79% for nicotine, and 62% for tar. CONCLUSIONS: Compensation, demonstrated when switching from a high tar cigarette to a low tar one, was incomplete. Advising people who have failed to give up smoking to switch to low tar cigarettes will reduce the intake of smoke constituents to a small extent. This would be expected to decrease their risk of smoking-related diseases, although by a smaller amount than would be achieved by giving up smoking altogether. PMID- 7491551 TI - Comparison of fluticasone propionate and beclomethasone dipropionate on direct and indirect measurements of bronchial hyperresponsiveness in patients with stable asthma. AB - BACKGROUND: Fluticasone propionate is a new inhaled corticosteroid with a 2:1 efficacy ratio compared with beclomethasone dipropionate with regard to lung function and symptom scores, without increased systemic activity. The aim of this study was to investigate whether this was also the case for bronchial hyperresponsiveness, assessed by both a direct (histamine) and an indirect (ultrasonically nebulised distilled water (UNDW)) provocation test. METHODS: Fluticasone propionate, 750 micrograms/day, and beclomethasone dipropionate, 1500 micrograms/day, were compared in a randomised, double blind, crossover study consisting of two six week treatment periods, each preceded by a three week single blind placebo period. Twenty one non-smoking asthmatics (mean forced expiratory volume in one second (FEV1) 74.7% predicted, mean PC20histamine 0.36 mg/ml) completed the study. RESULTS: Fluticasone propionate and beclomethasone dipropionate improved FEV1, peak flow rates, asthma symptoms, and bronchial hyperresponsiveness to the same extent. Both fluticasone propionate and beclomethasone dipropionate caused an increase in PC20histamine (mean 2.29 [95% confidence interval 1.45 to 3.13] and 1.95 [1.07 to 2.84] doubling doses, respectively) and in PD20UNDW (1.12 [0.55 to 1.70] and 1.28 [0.88 to 1.70] doubling doses, respectively). Neither treatment changed morning serum cortisol levels, but fluticasone propionate decreased the number of peripheral blood eosinophils less than beclomethasone dipropionate, indicating smaller systemic effects of fluticasone propionate. CONCLUSIONS: These findings show that fluticasone propionate is as effective as twice the dose of beclomethasone dipropionate on bronchial hyperresponsiveness, assessed by provocation with both histamine and UNDW, without increased systemic activity. PMID- 7491552 TI - Short-term association between air pollution and emergency room visits for asthma in Barcelona. AB - BACKGROUND: Several studies have assessed the association between urban air pollutants and hospital admissions or emergency room visits for asthma with inconsistent results. The objective of this study was to assess the relation between levels of black smoke, sulphur dioxide, nitrogen dioxide, and ozone and adult emergency room visits for asthma in Barcelona, Spain during the five year period 1985-9. METHODS: The daily number of emergency room visits for asthma was obtained from a register of respiratory emergencies designed to study the asthma outbreaks occurring in Barcelona. The association between asthma visits and levels of pollutants was assessed separately for summers and winters with Poisson regression models controlling for meteorological and time related variables. RESULTS: Black smoke was associated with asthma visits in summer but not in winter. The relative risk (RR) of asthma visits for a 25 micrograms/m3 increase of current day concentrations of black smoke was 1.082 (95% CI 1.011 to 1.157). The mean current and previous three day levels of black smoke led to a stronger association (RR = 1.114 (95% CI 1.010 to 1.160). In addition, nitrogen dioxide was associated with asthma visits in both summer (RR = 1.045, 95% CI 1.009 to 1.081) and winter (RR = 1.056, 95% CI 1.011 to 1.104). These associations were slightly higher for the previous day's level of nitrogen dioxide. No associations were found for sulphur dioxide or for ozone. CONCLUSIONS: This study provides further evidence of the effect of particulate pollution on asthma, and it suggests that nitrogen dioxide may have a role in the exacerbation of bronchial asthma in adults. PMID- 7491553 TI - Exacerbations of asthma without sputum eosinophilia. AB - BACKGROUND: Sputum analysis provides a non-invasive method of examining the airway secretions of subjects with asthma in order to better understand the inflammatory process. Increased proportions of eosinophils are generally seen in the sputum of subjects with asthma, especially when there is an exacerbation. An unexpected observation in the sputum of subjects with mild exacerbations of asthma is reported. METHODS: Thirty four consecutive subjects with symptoms consistent with a mild exacerbation of asthma were recruited for a treatment study. Inclusion criteria required persistent symptoms of chest tightness, dyspnoea, or wheezing for two weeks (without spontaneous improvement or alteration in dose of inhaled corticosteroid) and a forced expiratory volume in one second (FEV1) that was reversible to more than 75% predicted or known best to ensure the exacerbation was mild. Sputum (spontaneous or induced with hypertonic saline) from all subjects was examined for differential cell counts. Eosinophilic sputum was defined as > or = 4% eosinophils on two occasions or > 10% eosinophils once. Clinical characteristics, sputum differential counts, and measurements of airways obstruction were compared between the subjects with and without sputum eosinophilia. RESULTS: Almost half of the subjects (16 of 34) considered to have mildly uncontrolled asthma had no sputum eosinophilia. In comparison with the subjects who had sputum eosinophilia the non-eosinophilic group had less airways obstruction (FEV1% predicted 88% v 70%) and less severe airways hyperresponsiveness (PC20 methacholine 0.45 mg/ml v 0.13 mg/ml). There was no difference between the groups in the type or prevalence of symptoms, history of recent infections, smoking, relevant allergen exposure, or use of inhaled corticosteroid. CONCLUSIONS: Symptoms of mildly uncontrolled asthma are not always associated with eosinophilic airways inflammation as measured by sputum analysis. The causes and treatment of the non-eosinophilic condition require further investigation. PMID- 7491554 TI - Abnormal myocardial repolarisation in response to hypoxaemia and fenoterol. AB - BACKGROUND: Prolongation of the QTc interval has been associated with cardiac dysrhythmias and sudden death. QTc dispersion (interlead variability in QTc interval) has recently been proposed as being a more sensitive marker of repolarisation abnormalities and shown to be a more specific index of arrhythmia risk. Although hypoxaemia and fenoterol have previously been shown to prolong the QTc interval, this does not reflect regional myocardial repolarisation abnormalities. METHODS: Electrophysiological effects were measured at baseline and after 30 minutes steady state hypoxaemia at an arterial oxygen saturation (SaO2) of 75-80% (study 1) and at baseline then 30 minutes after inhaled fenoterol 2.4 mg (study 2). From the ECG, lead II corrected QT interval (QTc) and overall corrected QT dispersion were measured using a computer linked digitising tablet according to standard criteria. RESULTS: QTc dispersion was increased during hypoxia compared with baseline values (mean (SE) 69 (6) ms v 50 (5) ms) and after fenoterol compared with baseline (79 (13) v 46 (4) ms), respectively. There was also an increase in QTc interval and heart rate after fenoterol (493 (23) v 420 (6) ms and 98 (3) v 71 (6) bpm, respectively). The heart rate was increased during hypoxaemia compared with baseline (78 (3) v 64 (2) bpm), but no change occurred in the QTc interval. CONCLUSIONS: Both hypoxaemia and fenoterol cause myocardial repolarisation abnormalities in man in terms of increased QTc dispersion, but only fenoterol increased the QTc interval. This may be relevant in the aetiology of arrhythmias in patients with acute severe asthma where beta agonist therapy and hypoxaemia coexist. PMID- 7491555 TI - Association of non-wheezing lower respiratory tract illnesses in early life with persistently diminished serum IgE levels. Group Health Medical Associates. AB - BACKGROUND: The role of lower respiratory tract illnesses (LRIs) in the development of allergies is not well understood. The relation of wheezing and non wheezing LRIs to serum IgE levels and atopy was studied in 888 children. METHODS: Total serum IgE levels were measured at birth, nine months and six years of age; and interferon gamma production by blood mononuclear cells was measured at birth and nine months. Atopy was determined by skin prick tests at age six. Wheezing and non-wheezing LRIs up to age three were diagnosed by a physician. RESULTS: Cord serum IgE levels were similar between all LRI groups and the no LRI group. Children who had wheezing LRIs until the age of three had IgE levels at nine months and at six years within normal ranges for age. In contrast, children who had a non-wheezing LRI before the nine month IgE sample had lower IgE levels at nine months and six years (geometric mean 1.8 IU/ml and 9.9 IU/ml, respectively) compared with children who had no LRIs (3.9 IU/ml and 38.3 IU/ml, respectively). Children who had non-wheezing LRIs after the nine month IgE sample had normal nine month IgE levels (3.2 IU/ml) but decreased IgE levels at six years of age (15.7 IU/ml). Children with more than one non-wheezing LRI before the age of three were less likely to be atopic than those with no LRI (odds radio 0.2). Interferon gamma production was higher in the non-wheezing LRI group at nine months than in the no LRI or wheezing LRI groups. CONCLUSIONS: Non-wheezing LRIs are associated with subsequent depression of IgE levels and reduced skin test reactivity. PMID- 7491556 TI - Role of elevated plasma soluble ICAM-1 and bronchial lavage fluid IL-8 levels as markers of chronic lung disease in premature infants. AB - BACKGROUND: Pulmonary neutrophilia characterises both the relatively transient inflammation associated with infant respiratory distress syndrome (IRDS) and the persistent inflammation of chronic lung disease. The possibility that persistently raised markers of inflammation indicate the development of chronic lung disease in low birth weight (< 1730 g) preterm (< 31 weeks) infants was therefore investigated. METHODS: Soluble ICAM-1 (sICAM-1) levels in plasma, and interleukin (IL)-8 and myeloperoxidase (MPO) levels in bronchial lavage fluid (BLF) obtained from 17 infants on days 1, 5, and 14 following birth were measured and correlations with the number of neutrophils in BLF sought. Peripheral neutrophils were isolated on Polymorphoprep and chemotactic responsiveness to IL 8 was assessed using micro Boyden chambers. RESULTS: Sixteen infants developed IRDS and, of these, 10 infants subsequently developed chronic lung disease. Levels of IL-8 in BLF at 14 days of age correlated with the long term requirement for intermittent positive pressure ventilation (IPPV). Interleukin 8 levels in BLF correlated with neutrophil numbers and MPO concentration, suggesting both recruitment and activation in response to this cytokine. Antibody depletion studies showed that approximately 50% of total neutrophil chemotactic activity in BLF was due to IL-8. No difference in peripheral neutrophil chemotactic responsiveness at any age was observed for infants with IRDS or chronic lung disease. Plasma soluble intercellular adhesion molecule (sICAM-1) was higher at 14 days of age in infants who developed chronic lung disease than in those with resolving IRDS, and correlated with severity of disease, as indicated by duration of IPPV. CONCLUSIONS: The results indicate that high levels of plasma sICAM-1 and IL-8 in BLF at day 14 correlate with the development of chronic lung disease and indicate the severity of disease. PMID- 7491557 TI - Comparative effects of plasma exchange and pyridostigmine on respiratory muscle strength and breathing pattern in patients with myasthenia gravis. AB - BACKGROUND: Pyridostigmine, an acetylcholinesterase antagonist, is useful in improving respiratory function in patients with myasthenia gravis. More recently, plasma exchange has been employed in myasthenia gravis because it acts presumably by removal of circulating antibodies against acetylcholine receptors. Surprisingly, comparative data on the effects of pyridostigmine and plasma exchange on lung volumes, respiratory muscle strength, and ventilatory control system in patients with myasthenia gravis are lacking. METHODS: Nine consecutive patients with grade IIb myasthenia gravis were studied under control conditions and after a therapeutic dose of pyridostigmine. In a second study the patients were re-evaluated a few days after a cycle of plasma exchange, before taking pyridostigmine. In each subject pulmonary volumes, inspiratory (MIP) and expiratory (MEP) muscle force, and respiratory muscle strength, calculated as average MIP and MEP as percentages of their predicted values, were measured. The ventilatory control system was evaluated in terms of volume (tidal volume, VT) and time (inspiratory time, TI, and total time, TTOT) components of the respiratory cycle. Mean inspiratory flow (VT/TI)--that is, the "driving"--and TI/TTOT: that is, the "timing"--components of ventilation were also measured. RESULTS: In each patient treatment relieved weakness and tiredness, and dyspnoea grade was reduced with plasma exchange. Following treatment, vital capacity (VC) increased on average by 9.7% with pyridostigmine and by 14% with plasma exchange, and MIP increased by 18% and 26%, respectively. In addition, with plasma exchange but not with pyridostigmine forced expiratory volume in one second (FEV1) increased by 16% and MEP increased by 24.5%, while functional residual capacity (FRC) decreased a little (6.8%). The change in respiratory muscle strength was related to change in VC (r2 = 0.48). With plasma exchange, VT increased by 18.6% and VT/TI increased by 13.5%, while neither TI nor TI/TTOT changed. CONCLUSIONS: Plasma exchange can be used in patients with myasthenia gravis when symptoms are not adequately controlled by anticholinesterase agents. Plasma exchange increases respiratory muscle force and tidal volume due to changes in "driving" but not "timing" of the respiratory cycle. PMID- 7491558 TI - Relationship between cells obtained by bronchoalveolar lavage and survival in idiopathic pulmonary fibrosis. AB - BACKGROUND: The relationship between cell types in bronchoalveolar lavage (BAL) fluid and the clinical course of patients with idiopathic pulmonary fibrosis (IPF) has been the subject of several studies. However, the results of these studies are not conclusive. The aim of this study was to investigate the relationship between the absolute and relative cell numbers in BAL fluid from patients with IPF and their survival. METHODS: Results obtained from the initial BAL fluid analyses of all histologically proven cases of IPF (n = 49) were selected retrospectively. Cox's proportional hazards survival analysis was used for estimating the relationship between absolute and relative cell numbers and survival. RESULTS: A negative relationship was found between both the absolute numbers and percentages of eosinophils in BAL fluid samples and survival. No such relationship was demonstrated for the absolute numbers or the percentages of any other cell type. CONCLUSIONS: Although this study has restrictions, these findings suggest a negative relationship between the absolute numbers and percentages of eosinophils in BAL fluid samples and survival in patients with IPF. PMID- 7491559 TI - Streptococcus milleri pulmonary disease: a review and clinical description of 25 patients. AB - BACKGROUND: Streptococcus milleri is increasingly being recognised as an important pulmonary pathogen which may lead to the development of empyema or lung abscess. Although several small series have been reported, the clinical and laboratory features have yet to be fully characterised. METHODS: Twenty five cases were identified and the clinical and laboratory data from case records were analysed. RESULTS: There were 16 empyemas, five lung abscesses, and four with both lung abscess and empyema. The mean age of the patients was 61 years (range 36-89) and 84% were men. The most common symptoms at presentation were shortness of breath, chest pain, cough, and weight loss; only 36% had a fever. Four of the nine patients with lung abscess required a diagnostic lobectomy because of suspected malignancy. Predisposing factors were present in 80% of patients and included the following: pneumonia, periodontal disease, excess alcohol intake, previous thoracic surgical procedures, and malignancy. Laboratory features of S milleri infection were leucocytosis, neutrophilia, anaemia, abnormal liver function tests, and hypoalbuminaemia. In the group with empyema five patients had a pneumothorax on initial presentation and pleural loculation occurred in 10 of these patients. The median stay in hospital was 34 days (range 11-88). Six patients died, five of whom had significant underlying illnesses. CONCLUSIONS: Pulmonary infection with S milleri may result in considerable morbidity and mortality, and is characterised by a strong male predominance, non-specific symptoms (often without toxicity), the presence of predisposing factors, pleural loculation, pneumothorax, and a protracted stay in hospital. PMID- 7491560 TI - Cardiopulmonary exercise testing following bilateral thoracoscopic sympathicolysis in patients with essential hyperhidrosis. AB - BACKGROUND: Essential hyperhidrosis is characterised by an overactivity of the sympathetic fibres passing through the upper dorsal sympathetic ganglia D2-D3. Anatomical interruption at the D2-D3 level is a highly effective treatment for essential hyperhidrosis but also causes (partial) cardiac denervation and, after surgical sympathicolysis, important impairment of cardiopulmonary exercise function has been observed. The purpose of this study was to compare the results of cardiopulmonary exercise testing between patients with essential hyperhidrosis and a normal control population, and to examine the effects of thoracoscopic D2 D3 sympathicolysis on cardiopulmonary exercise capacity in patients with essential hyperhidrosis. METHODS: maximal, symptom limited incremental exercise tests were performed in 26 patients with severe essential hyperhidrosis one week before and one month after D2-D3 thoracoscopic sympathicolysis, and in 14 age and sex matched healthy volunteers. D2-D3 thoracoscopic sympathicolysis was performed using a simplified one stage bilateral procedure. RESULTS: Palmar hyperhidrosis was relieved in every patient, confirming the D2-D3 denervation. A higher peak heart rate (7%) was seen in the patient group than in the normal subjects, but ll other cardiovascular, metabolic, and respiratory parameters were similar. After D2-D3 thoracoscopic sympathicolysis, heart rate at rest (13%) and at peak exercise (7%) were reduced, together with an increase in oxygen pulse. All other parameters remained unchanged. CONCLUSIONS: Sympathetic overactivity relevant to cardiovascular function in essential hyperhidrosis is evident only during sympathetic stimulation. D2-D3 thoracoscopic sympathicolysis causes a small and asymptomatic reduction in maximal and resting heart rate and is not associated with a decrease in exercise capacity, in contrast with the detrimental effects on exercise capacity of open surgical sympathectomy. PMID- 7491561 TI - Repeaters count: a sentinel method for asthma outbreaks. Barcelona Soybean-Asthma Group. AB - BACKGROUND: Asthma outbreaks have not been detected in Barcelona, Spain since the epidemic of 1981-7. On 29 October 1994 several subjects attended a Barcelona hospital because of acute attacks of asthma and were identified as former soybean epidemic asthma patients (repeaters), raising the possibility of an asthma soybean-related episode. The usefulness of counting repeaters to evaluate small increases of acute asthma in a non-epidemic period is illustrated. METHODS: A retrospective identification of asthma admissions was performed, pollution filters collected before and after the index day were analysed for contents of low molecular weight soybean allergen, and soybean unloading activities were investigated. RESULTS: There was no epidemic increase of asthma patients during this day in the whole of Barcelona, but an unusually high number were repeaters (seven observed v 0.483 expected). A relationship with increased levels of low molecular weight soybean allergen (U/m3) was detected in the available pollution filters collected the day before and after the index day. Two ships were unloading soybean in the city harbour on the index day. CONCLUSION: Soybean unloading at the harbour of Barcelona released soybean dust and produced a small cluster of emergency room visits for asthma in patients formerly affected by soybean asthma epidemics. It is suggested that counting repeaters could provide the basis for a systematic epidemiological surveillance of sensitised populations. PMID- 7491562 TI - Review of the prescription of domiciliary long term oxygen therapy in Scotland. AB - BACKGROUND: Since 1989 long term oxygen therapy (LTOT) in Scotland has been prescribable only by respiratory physicians, whereas in England and Wales general practitioners can also prescribe this treatment. The effect of this policy has been audited. METHOD: Six hundred and thirty patients were prescribed LTOT in Scotland between 1 October 1989 and 30 September 1991, of which 519 case notes were reviewed. RESULTS: In 79% of patients the diagnosis was chronic obstructive pulmonary disease (COPD), with a near equal male to female ratio. The mean (SD) age was 65 (13) years (range 0.2-90). Sixty percent of patients died during the study period. Compliance with treatment was 14.9 (6.0) hours daily (range 1-24) and in 44% was less than 15 hours daily. Sixty one percent were clinically unstable when assessed and 14% were still smoking. Only 14% of those with COPD fulfilled all of the relative criteria for the prescription of LTOT. Only 56% had a repeated arterial blood gas measurement within 12 months of prescription and 51% of these were taken during a period of clinical instability. Expenditure on oxygen cylinders was six times greater than on oxygen concentrators over this period. CONCLUSIONS: Compliance with LTOT in Scotland, where prescription is the responsibility of respiratory physicians, is similar to other studies and ranges widely. The greatest problem concerning adherence to the guidelines is assessment during clinical instability. The number of deaths suggest that prescription occurs late in the course of the disease. This study highlights the areas where the prescription of LTOT needs to be improved. PMID- 7491563 TI - Sleep-related breathing disorders. 5. Nasal continuous positive airway pressure treatment for obstructive sleep apnoea. AB - CPAP should be considered the first line of treatment in patients with moderate to severe obstructive sleep apnoea. In our centre in Sydney this generally means patients with more than 20 apnoea/hypopnoeas per hour with repeated dips in oxyhaemoglobin saturation and usually some symptomatology. Despite this first line role of nasal CPAP, recent objective studies question whether earlier enthusiastic reports on adherence to CPAP are correct. The role of technical innovations in new CPAP machines in improving usage remains to be tested. The "drop out" rate from physician selection for a CPAP trial to highly compliant user is certainly more than 50% of patients. What happens to these patients? Data from some studies suggest that surgical treatments are used, at least in the USA, but in all probability many of these patients remain untreated. The challenge in the next decade is either to improve CPAP devices to increase usage in this group or to develop other treatment options. The role of intensive inhospital "acclimatisation" to CPAP also has yet to be objectively tested. It is unclear whether "intelligent" CPAP will make huge inroads in increasing the number of patients who accept CPAP trials, prescriptions, or compliance. It will have minimal impact on patients with mask problems or claustrophobia or those who feel that CPAP is inconvenient. There is a high likelihood that it will reduce technologist workload during CPAP titration studies. "Intelligent" CPAP may help to reduce total overnight mouth leakage and therefore reduce nasal side effects. The current expense of developing such devices will mean that they are unlikely to supersede much cheaper standard "one pressure" CPAP machines in the next few years. PMID- 7491564 TI - Fallibility of transthoracic needle biopsy of anterior mediastinal masses. AB - Percutaneous transthoracic core needle biopsy has been advocated as a highly accurate technique for the diagnosis of anterior mediastinal masses. A patient is described with a large anterior mediastinal mass in whom the diagnosis of mediastinal carcinoid tumour was made by transthoracic core needle biopsy. At definitive surgical resection the tumour proved to be a B cell lymphoma. This case illustrates one of the important limitations of needle biopsy with its potential for sampling error. PMID- 7491566 TI - Management of residual thymic cysts in patients treated for mediastinal Hodgkin's disease. AB - The pathogenesis of residual thymic cysts after treatment for mediastinal Hodgkin's disease is uncertain. Their presence after adequate treatment often presents the oncologist and the thoracic surgeon with a therapeutic dilemma. Two patients with residual thymic cysts after curative treatment for mediastinal Hodgkin's disease are described and the management discussed. PMID- 7491565 TI - Recall lung pneumonitis due to carmustine after radiotherapy. AB - A patient who developed pneumonitis immediately after the administration of carmustine (BCNU), within exactly the same field as previous irradiation, is presented. The patient responded partially to corticosteroids. This case suggests that irradiation causes subclinical sensitisation of the lung and can therefore have an additive effect in precipitating lung damage when another pulmonary toxin is encountered at a later date. PMID- 7491570 TI - Guidelines for care during bronchoscopy. PMID- 7491568 TI - Commentary: lymphoma involving the mediastinum--challenges in diagnosis and management. PMID- 7491569 TI - Apnoea in Duchenne muscular dystrophy. PMID- 7491567 TI - Lethal tracheal dissolution during treatment for thyroid lymphoma. PMID- 7491571 TI - Chest physiotherapy in cystic fibrosis. PMID- 7491572 TI - [Tuberculosis is moving]. PMID- 7491574 TI - [Intervention cardiology. Atherectomy or stenting?]. PMID- 7491573 TI - [Perfume allergy--an allergy of prosperity?]. PMID- 7491575 TI - [Atherectomy. A new catheterization method for coronary revascularization]. AB - Intimal atheroma con be excised and removed by coronary atherectomy. We describe our experience from the use of this method in the first 42 patients. The procedure was successful in 40 patients, in 16 cases after adjunct percutaneous transluminal coronary angioplasty. Two patients required bypass surgery because of catheter-related injury to the coronary vessel. One of these suffered a myocardial infarction, the only infarction in the material. In-hospital complications were minor, and the average stay in hospital after the procedure was 1.6 days. Angiography in 20 patients after a median follow-up period of 116 days showed restenosis in 30%, all of whom were treated successfully with PTCA. Symptomatic improvement was reported by 74% of the patients after a median observation time of 16 months. One patient died, probably from infarction, three months after the initial treatment, one patient suffered a non-fatal infarction, and one patient underwent bypass surgery. We conclude that atherectomy is an effective and safe treatment of coronary heart disease in selected cases, mainly eccentric stenoses and ostial lesions. PMID- 7491577 TI - [Extrapulmonary tuberculosis. An important differential diagnosis in immigrants with suspected malignancy]. AB - The incidence of tuberculous disease is increasing all over the world, mostly in the poor, developing countries, but also in some industrialized countries. In Norway, extrapulmonary tuberculosis is a rare phenomenon. It is found mostly among older Norwegians and in younger immigrants from the third world. Since the disease is rare, it may be overlooked or confused with malignant disease. We describe two patients with unusual forms of extrapulmonary tuberculosis, both mimicking neoplastic disease. The first patient was a 27-year-old woman from South-East Asia, who was operated on for suspected intraductal comedo-type carcinoma of the breast, but histological examination showed tuberculous mastitis. The second patient was a 26-year-old man from East Africa with a medical history indicating intra-abdominal lymphoma. The final diagnosis, however, was mesenteric tuberculous lymphadenitis. Both patients were treated successfully with isoniazid, rifampicin and pyrazinamide. PMID- 7491578 TI - [Contact allergy to essential oils]. AB - Ethereal oils are widely used, in food, toothpaste, as flavouring agents in perfumes and cosmetics, and in dermatological treatment, as antiseptics, adstringenta, antipruritic tinctures, lotions and pomades. The use of ethereal oils seems to have increased in recent years, probably due to a growing interest in alternative medicine, and especially aromatherapy. By describing the cases of three patients with allergic contact dermatitis, we wish to point out possible unwanted side effects of using ethereal oils. PMID- 7491576 TI - [Tuberculosis among AIDS patients in Ullev]l hospital]. AB - Records of patients with concomitant HIV infection and human tuberculosis were analysed. Nine out of 232 AIDS patients (4%) developed human tuberculosis with a preponderance (6/9) of immigrants from sub-Saharan Africa. In three patients the diagnosis was delayed because of atypical manifestations of the disease and lack of typical chest X-ray findings. Tuberculin skin tests were positive in only three patients, and became negative in one patient who developed two episodes of tuberculosis. All the patients who complied with the conventional triple anti tuberculosis regimen responded well, and no multidrug-resistant tuberculosis was observed. PMID- 7491581 TI - [Unstable angina pectoris. Pathogenesis, treatment and prognosis]. AB - Unstable angina pectoris is a term used for various clinical conditions of angina pectoris, such as new onset of angina, accelerated angina, resting angina and post-infarction angina. The natural history varies according to clinical presentation, and the prognosis differs considerably for the different types of unstable angina. The first step in the pathophysiological mechanisms leading to unstable angina is thought to be rupture of an atherosclerotic plaque. Exposure of the content of the plaque leads to varying degrees of coronary obstruction, due to coronary thrombosis and vasoconstriction. A key element is dysfunction and activation of the endothelium. Patients with unstable angina should be hospitalized and treated with anti-anginal drugs, and with aspirin and heparin to reduce risk of myocardial infarction. Coronary angiography should be considered to identify patients who might benefit from angioplasty or coronary bypass surgery. Reduction of risk factors is important for long-term outcome. PMID- 7491580 TI - [Adult patients with atrial septal defect]. AB - Our material included patients older than 30 years who were diagnosed as having atrial septal defect during the period 1980-1993. A questionnaire was used during follow-up. 39 of a total of 42 patients were operated on. The mean follow-up time was 5.5 years. There was no operative mortality. Five patients had died by the time of follow-up. The three patients who had not been operated on had all died. The available literature gives no conclusive answer to the question of whether all patients with an atrial septal defect should be offered surgery or not. PMID- 7491579 TI - [Chemotherapy-induced hemolytic uremic syndrome. A complication to mitomycin treatment of breast carcinoma]. AB - Mitomycin is a cytotoxic antibiotic that was first introduced into clinical use in 1958. Not until twenty years later was it recognised that haemolytic uremic syndrome could develop after treatment with mitomycin. It can be asked whether this condition was undiagnosed in previous years, since its frequency is now reported to be 4-15%. The disease appears to be dose-related, since it rarely occurs in patients who have received mitomycin < 30 mg/m2. No effective therapy has been established. We describe two patients with breast cancer in remission after treatment with mitomycin in combination with 5-fluorouracil. Both developed haemolytic uremic syndrome with fatal outcome. PMID- 7491582 TI - [The end of antibiotics? Increasing bacterial resistance in global and Norwegian perspective]. AB - During the last 20 years there has emerged a growing world-wide problem with regard to multidrug-resistant microbes. The most serious examples so far are vancomycin-resistant strains of Enterococcus faecium, totally resistant isolates of Mycobacterium tuberculosis and multiple-resistant Staphylococcus aureus and Streptococcus pneumoniae. With the exception of some few strains of methicillin resistant S. aureus and vancomycin-resistant enterococci, such bacteria have not been found in Norway. In this article we discuss possible ways of preventing further selection and spread of multiple-resistant microbes. We stress the importance of infection control programmes and restrictive use of antibiotics. PMID- 7491583 TI - [Revolution in diagnostic microbiology. Needs, driving forces and consequences]. AB - Introducing molecular biological technology into medical microbiology promotes deeper insight into the epidemiology, pathogenesis, diagnostics and treatment of infectious diseases. New technology has helped to highlight the complicated biological interaction between host and microbes and has created a need for more advanced technology both in microbiological diagnostics and in microbiological science. Extra-professional factors like the high prestige of new techniques have also been of considerable significance. Medical microbiologists will become an important link between the inventors of new technology and the physicians who diagnose and treat the patients. Through these contacts and their diagnostic laboratory work, they will become important generators and communicators of knowledge. In this connection it will be important to represent reason in introducing technology in medical microbiological diagnostics. Since patients are expected to play a more active part in health care, communicating knowledge to patients and the population in general will become an obligation. PMID- 7491584 TI - [Tuberculosis trends in Norway--has the decline stopped?]. AB - The aim of the study was to present the trend as regards tuberculosis in Norway since 1950, on the basis of data on incidence and mortality in the total population and infection rate among pupils leaving primary school. The decline in incidence continued until the late 1980s. While the decline was 11% per year during the period 1950-74, it has been slower in recent years. In 1990-92 the incidence level was somewhat higher than in 1989, but figures for 1993-94 indicate a new decline. Mortality declined more slowly after the mid-1970s; a similar trend is found for prevalence of infection among pupils. The incidence is among the lowest in Europe. The proportion of cases among immigrants increased gradually from 4% in 1977 to 40% in 1993. Among the Norwegian-born population the incidence rate of infectious pulmonary tuberculosis has shown a steady decline, also during the last decades. Norwegian-born cases are mainly elderly persons infected many decades ago. New infection is probably rare, and the proportion of infected persons is gradually declining. PMID- 7491585 TI - [Health care personnel in Norway]. AB - In 1990, the Nordic Council of Ministers initiated the joint Nordic project on "Work and health among health care personnel". The main aims were: To review and evaluate research data concerning the health and work of health care personnel in the Nordic countries, initiate joint Nordic projects and promote collaboration between Nordic researchers. Altogether more than one million people in the Nordic countries are employed in the health care sector, or almost 10% of the labour force. In all the Nordic countries the labour force in the health sector is dominated by women; the proportion of women being between 84 and 87%. In Norway and Sweden a large share of the labour force works part time. When controlled for sex and level of education, sick leave is the same among health personnel as among the general working population. As in the whole population, sick leave is higher among women, and among persons with a lower level of education. In general, workers in the health care sector in the Nordic countries run no greater risk of developing occupationally related injuries than other workers do. In a register-based study of Swedish workers it was found that the risk of being absent from work because of violence or threats is higher among health personnel than in the general working population. PMID- 7491586 TI - [Cultural variations of view on differences between the psyche and the soma]. PMID- 7491587 TI - [Frustration and confidence failure]. PMID- 7491588 TI - [Repetition and private examination--a way to medical studies]. PMID- 7491589 TI - [Chronic fatigue syndrome]. PMID- 7491590 TI - [Incomprehensible about science and communication]. PMID- 7491591 TI - [Home monitoring does not prevent crib death]. PMID- 7491592 TI - [Should moderate hypertension be medically treated?]. PMID- 7491593 TI - [When a patient does not hear]. PMID- 7491594 TI - [Radiographic diagnosis of pulmonary embolism]. PMID- 7491595 TI - [Hearing loss in children--failing diagnosis]. PMID- 7491596 TI - [Prone position and crib death]. PMID- 7491597 TI - [Crib death and infant mortality in the Nordic countries 1988-93]. AB - During the years 1970-89 there was a marked increase in cot deaths in the Nordic countries Denmark, Finland, Norway and Sweden. Since 1990 cot death has decreased dramatically, especially in Norway and Denmark. Mortality rates have decreased by 74% between 1989 and 1993 in Norway and by 69% between 1991 and 1993 in Denmark. Investigations in Norway during the same period have shown a change in the sleeping positions of infants, from prone positions to supine/side positions. Infant mortality rates have decreased in all the Nordic countries and in 1993 varied from 5.45 per thousand live births in Denmark to 4.40 in Finland. PMID- 7491598 TI - [Hearing of small children. Satisfactory testing at health centers?]. AB - Of 803 four-year-old children tested, audiometric screening detected ten with secretory otitis media necessitating operation and one child with sensorineural loss of hearing. Control showed audiometer calibrations and noise levels during screening to be within acceptable limits. The anamnestic information ought to be improved by asking direct questions about hearing disabilities at all age levels as part of the screening procedure. The one child with sensorineural hearing loss should have been detected earlier. Perhaps we place too much trust in the methods used to test young childrens' hearing. It may be wise to look more closely into the methods and routines used to screen for hearing defects at one year of age. PMID- 7491599 TI - [A rapid analysis of the lipid profile. A comparative study of different analytical methods for determination of blood lipids]. AB - A simple and rapid analytical assay for determination of total cholesterol, HDL cholesterol and triglycerides in capillary blood samples has been compared with established laboratory methods in a group of 41 test subjects. The new method produces results that are fully acceptable for determination of these three lipid parameters, and agree well with the results obtained in serum by established laboratory methods. The Cholestech LDX analyser may be useful for providing rapid and accurate measurements of the lipid profile, including total cholesterol, triglycerides and HDL-cholesterol. Its small size, the speed of analysis and the fact that it is easy to use make the Cholestech LDX a suitable instrument for use in out-patient clinics and primary care medicine. PMID- 7491600 TI - [Intellectual and psychosocial functions in children treated with radiation for brain tumor]. AB - Modern treatment has increased the survival rate for children with brain tumour, but treatment with radiation may lead to late adverse neuropsychologic effects. Also, peer problems may follow long periods away from school, and painful treatment and anxiety about the disease may mobilize emotional stress reactions. 16 brain tumour children with a mean age of 11 years (range 6-16 years), all of them treated with radiation, were tested with WISC-R. The mean time between diagnosis and study was five years (range 2-11 years). Parents and teachers completed the Achenbach check lists CBCL and TRF. Children below age four years at diagnosis achieved the lowest WISC-R IQs, the highest problem scores and the lowest social adaptive scores on CBCL and TRF. More problems were reported among the youth group aged 12-16 years than among the child group aged 6-11 years. Hypoactivity and low psychomotor speed seemed to have an adverse effect on both intellectual and psychosocial functioning. An almost normal communicative and reasoning ability was a resource for most of the children. PMID- 7491601 TI - [Nitrogen oxide in the treatment of oxygenation failure in neonates]. AB - Oxygenation problems are common in very sick infants, and involve a high risk of mortality or residual morbidity. We describe two infants with severe oxygenation difficulties. In one baby these were caused by meconium aspiration, while in the second, premature (1,500 g birth weight) infant Escherichia coli septicemia had precipitated persistent pulmonary hypertension. Inhaled nitric oxide led to immediate improvement in oxygenation, and the one infant was weaned off mechanical ventilatory support after four days, the other after ten days. There were no discernible side effects in either baby. Nitric oxide treatment should be considered for infants with life-threatening oxygenation problems who do not respond adequately to conventional management. PMID- 7491602 TI - [Fibrodysplasia (myositis) ossificans progressiva]. AB - A seven-year-old boy with mental retardation and congenital skeletal malformations in the thumbs and big toes developed recurrent lumps in the shoulder and interscapular region. The lumps subsided slowly but left severe stiffness in the affected areas. The first biopsy revealed oedema and a chronic inflammatory response with fibromyxoid proliferation of the soft tissue. A second biopsy revealed ossification of muscle and fascia. The patient is a typical case of fibrodysplasia ossificans progressiva, an autosomal dominant inherited disorder characterized by congenital skeletal malformations in toes and fingers and progressive ectopic ossification, often combined with alopecia, deafness and in rare cases mental retardation. The disorder leads to severe physical disability in young age and respiratory and nutritional problems. No effective treatment is known, but it is important to avoid exacerbating factors such as biopsy, operations and intramuscular injections. PMID- 7491603 TI - [Diagnosis and treatment of familial hypercholesterolemia in children and adolescents]. AB - The identification and management of hyperlipidemias in children are currently limited to persons regarded as being at very high risk, such as those with familial hypercholesterolemia. Universal screening of children is not recommended, but it is important to screen the subset of children and adolescents at highest risk, i.e. those with a family history of premature cardiovascular disease or familial hypercholesterolemia. Treatment of children and adolescents with hypercholesterolemia requires a multidisciplinary approach. Cholesterol lowering medication may be required in addition to dietary therapy in children with a history of premature coronary heart disease among close relatives. PMID- 7491604 TI - [Proper use of antibiotics--what is that?]. AB - Antibiotics are one of the cornerstones of modern medicine. During the last 20 years there has been an alarming world-wide spread of multiple-resistant bacteria. One of the main reasons is the overuse of all types of antibiotics, especially broad-spectrum drugs. This paper gives general advice on antibiotic therapy in Norway. We adovcate the use of drugs with little ecological impact, such as the penicillins. Whenever possible, empiric treatment with macrolides, tetracyclines cefalosporines, imipenem and fluoroquinolones should be avoided. PMID- 7491605 TI - [Infections in intensive care units. A challenge for the specialist of infectious diseases?]. AB - There is an obvious need for teamwork between different specialists on the diagnosis and treatment of patients in an intensive care unit. The infectious disease specialist must contribute by establishing definite and quick diagnosis of infections and by employing an adequate and sensible antibiotic policy for successful treatment. The local microbiological flora should be kept at an acceptable level of resistance to antibiotics, and preventive measures should be chosen carefully. To achieve this goal, extensive knowledge on antibiotics and microbial epidemiology is essential. PMID- 7491606 TI - [Spiral compter tomography in acute pulmonary embolism]. AB - 13 patients with clinically suspected acute pulmonary embolism were assessed by means of contrast-enhanced spiral volumetric computed tomography (CT). Our experience is already so positive that we wish to focus on this method. Spiral CT gives an exact diagnosis in many cases where perfusion scintigraphy is uncertain. In our opinion spiral CT can be reliably used in patients with clinically suspected pulmonary embolism, and appears to offer an alternative to scintigraphy and pulmonary angiography. PMID- 7491607 TI - [Familial hypercholesterolemia in children. A management program. Procedures for diagnosis and treatment in our pediatric departments]. AB - A Norwegian programme for treatment and selective screening of familial hypercholesterolaemia has been developed which takes into account family history and levels of hypercholesterolaemia. The programme includes recommendations on when and whom to screen for familial hypercholesterolaemia. With regard to treatment, special emphasis is placed on diet. The working group suggests that small lipid clinics with a dietitian should be established in some paediatric departments. PMID- 7491608 TI - [Change in infants' sleeping position and incidence of crib death in Northern Norway]. AB - Several investigations have reported a prone sleeping position to be a risk factor for sudden infant death syndrome (SIDS). We investigated whether changes in sleeping position among infants in Northern Norway were associated with changes in SIDS rates. The prevalence of prone sleeping position was 54% among infants born in the years 1976-1989 and 5% among infants surveyed in May 1994. During the same period the SIDS rate declined from 2.75/1000 to 1.57/1000. These findings are consistent with other research in the field, and indicate that a prone sleeping position is a risk factor for SIDS. PMID- 7491609 TI - [Is drug use in maternity wards baby-friendly enough?]. AB - Quality assurance in the care of breast-feeding women and their nursing infants also applies to drugs administered during delivery and puerperium. Large variations among hospitals may indicate that drug use is irrational. A survey comparing the extent of drug sales from the hospital pharmacy to maternity wards in eight Norwegian hospitals was performed in 1992 and the results were compared with data from 1988. The purpose was to examine whether the drug use was "baby friendly" with regard to the following criteria; proven efficacy for the indication; no effect on milk ejection, milk production and interaction with infant; minimal transfer of drug to milk. Large variations were found among hospitals in the case of some oxytocic drugs. High use of oxytocin as nasal spray and metylergometrine as tablets may indicate unnecessary use of drugs. A large decrease (89%) in the use of hypnotics was found from 1988 to 1992, which may indicate previous irrational use of these drugs. Pethidine as pain relief during delivery remained stable during this period, and was received by 40-60% of women giving birth. None of the drugs given to the mothers was assessed to represent a risk to the breast-fed infant. In general, drug use in maternity wards had decreased during the last four years and, with some exceptions, appeared to be more baby-friendly. PMID- 7491610 TI - [Disability pensions because of musculoskeletal diseases among women of different occupations]. AB - Altogether 9,516 women, 16-67 years of age, were granted a disability pension in Norway in 1993. Musculoskeletal disease was registered as the main diagnosis for 3,730. Employment status is known for 3,125 of these 3,730 women. Analyses, based on age standardized incidence rates--SIR's, pointed to shop assistants, practical nurses and charladies as the three major occupational groups with high risk of being disability pensioned because of musculoskeletal disease. These three occupations accounted for more than one third of all women in paid employment who were granted a disability pension for musculoskeletal disease 1993. Compensation will amount to about NOK 600 million before these women reach the normal retirement age of 67 years. Prevention of musculoskeletal diseases in the labour force would appear to be potentially cost-effective. So far, prevention of work related musculoskeletal diseases has focused mainly on the technical ergonomic aspects of the working conditions. It is recommended that more attention be given to organizational aspects, such as daily hours of work, the timing and duration of breaks, work pace and potential to influence one's own work situation. PMID- 7491611 TI - [Productivity in a Norwegian department of surgery]. PMID- 7491612 TI - [The risk of false positive Chlamydia tests should be empirically determinated]. PMID- 7491613 TI - [Instruction of medical students]. PMID- 7491615 TI - [Lipid diarrhea in young pigs]. PMID- 7491616 TI - [Toxicity of a flea pesticide]. PMID- 7491614 TI - [Cobalt supply to ruminants, a bottleneck?]. AB - On some dairy farms the supply of cobalt was expected to be adequate given the soil data and the composition of the ration. However cows on these farms produced less milk than expected and their condition was poor considering their ration and milk yield. The milk lactose content was less than 4.5%. The cows produced more milk after they were given extra cobalt (5.5 mg/day). The animals, condition improved, the milk lactose content rose, and the animals ate more dry matter than previously. A marked increase in the potassium content of the grass silage possibly depressed the availability of cobalt and caused vitamin B12 deficiency. This needs further investigation. PMID- 7491617 TI - [Poorly dilated cervix in ruminants]. PMID- 7491618 TI - [Annual congress 1995, its theme 'Animal and Art'. North Holland hold a mirror up to the veterinarian]. PMID- 7491619 TI - [142nd general meeting KNMvD (Royal Dutch Society for Veterinary Medicine). Executive Board: 'Quality care has to be!']. PMID- 7491621 TI - Ultrastructural study of osmolality effect on spermatozoa of three marine teleosts. AB - With particular emphasis on mitochondria that may provide endogenous energy for spermatozoan motility, the morphological changes of the spermatozoa of three marine teleosts, black porgy (Acanthopagrus schlegelli), black grouper (Epinephelus malabaricus), and Atlantic croaker (Micropogonias undulatus), were compared either after activation in artificial sea water or when immersed in various osmotic pressure media. The midpieces of these three teleosts spermatozoa are composed of mitochondria surrounding the flagellum. Each mitochondrion is enclosed by distinct outer and inner membranes. The inner membrane separates the organelle's volume into two phases: the matrix and the intermembrane space. The inner membrane displays numerous infolding cristae that vary in number and shape and extend into the matrix. Following activation with artificial sea water, spermatozoa became motile and both the size and number of mitochondria decrease and then totally disappear. The present study strongly suggests that an energy source(s), responsible for motility, is located within the mitochondria in the midpiece of these three marine teleost spermatozoa. PMID- 7491620 TI - Peroxisomes and sterol carrier protein-2 in luteal cell steroidogenesis: a possible role in cholesterol transport from lipid droplets to mitochondria. AB - In the present investigation, we have studied peroxisomes and sterol carrier protein-2 (SCP2) in control and luteinizing hormone stimulated rat luteal cells. Superovulated immature rats in mid-luteal phase (8 days after ovulation) were divided into two groups (n = 4/group) and treated with vehicle (0.2 ml saline), or luteinizing hormone (LH, 20 micrograms/rat). In this animal model, LH acutely stimulates steroidogenesis. Thirty minutes later, corpora lutea were fixed by whole body perfusion and processed for (1) electron microscopic immunocytochemistry to localize SCP2 via the protein A gold immunolabeling technique, and for (2) electron microscopic histochemistry to stain peroxisomal catalase via the alkaline 3,3'-diaminobenzidine tetrahydrochloride method. In the steroidogenic, mid-phase luteal cells of vehicle treated rats (controls), SCP2 was highly concentrated in peroxisomes and sparsely scattered on mitochondria, but no labeling was observed in lipid droplets. In the luteal cells of rats acutely stimulated with LH, peroxisomes immunolabeled for SCP2 were observed within the luteal cell lipid droplets and mitochondria, and in union with lipid droplets and mitochondria. Moreover, in contrast to control luteal cells, significant immunolabeling for SCP2 was detected within the lipid droplets and mitochondria in luteal cells of LH-treated rats. As SCP2 binds cholesterol to 1:1 molar ratio and is known to be involved in the intracellular movement of cholesterol, these findings suggest that peroxisomes and SCP2 may possibly be involved in delivering cholesterol from lipid droplets to the mitochondria when luteal cell steroidogenesis is acutely stimulated by LH. PMID- 7491622 TI - Expression of c-fos oncogenes in adrenal cortex of the rat with renal hypertension. AB - This study was conducted using immunohistochemistry to detect the expression of the prooncogen c-fos in the corticoadrenal of the rat with renal hypertension. A model of renal hypertension was established through narrowing a renal artery of the rat. An antiserum against FOS oncoprotein was used to immunohistochemically detect the activated cells of the corticoadrenal. In the experimental group, C FOS-immunoreactive (ir) nuclei proved to be present in the zona glomerulosa. Few labeled nuclei were seen in the lateral part of the zona fasciculata. An increasing number of C-FOS-ir nuclei was demonstrated in the intermediate part of the zona fasciculata. The highest concentration of the labeled nuclei was found in the inner part of the zona fasciculata. Scattered FOS-ir nuclei existed in the zona reticularis. The labeled nuclei were often round or oval in shape, and different in size. Only were a few labeled nuclei seen in the sham-operated group and no FOS-ir nuclei in normal control group. The result suggested that all three zones of the corticoadreal were modified subsequent to induction of renal hypertension. PMID- 7491623 TI - Subpopulations of mouse resident peritoneal macrophages fractionated on Percoll gradients show differences in cell size, lectin binding and antigen expression suggestive of different stages of maturation. AB - Aiming the identification of macrophage heterogeneity, mouse resident peritoneal cells were fractionated on discontinuous Percoll gradients into six discrete fractions (0, 1, 2, 3, 4 and 5 in order of increasing density). All six fractions and the total population were characterized by light and electron microscopy, and flow cytometry. The least dense fraction (0) had a low viability (44%); fractions 4 and 5 had a low percentage of macrophages. Light microscopy and flow cytometry of macrophage-enriched fractions 1, 2 and 3 showed an inverse correlation between diameter and cell density, as well as suitable differences in lectin binding to the macrophages of each fraction. The surface of macrophages from fraction 1 had more sialyl residues (higher binding of the lectin LFA), less galactosyl residues (lower binding of the lectin PNA) and expressed more receptors for the antibodies M1/70 (Mac-1) and F4/80; fraction 3 had an opposite binding behavior for the lectins and expressed fewer receptors for both antibodies; fraction 2 had an intermediate behavior for both parameters. Binding of the lectins Con A and HPA showed slight differences, whereas UEA I did not present a detectable difference among the fractions analyzed. These findings suggest that the macrophage heterogeneity achieved by the gradient separation of resident peritoneal cells could be explained by different stages of macrophage maturation. PMID- 7491625 TI - Freeze-fracture and deep-etched view of the cuticle of Caenorhabditis elegans. AB - At ultrastructural level, the Caenorhabditis elegans (C. elegans) cuticle shows the presence of well-defined layers, one of them is a membrane-like structure designated as epicuticle, always present on the outermost surface of nematodes. Freeze-fracture replicas revealed the existance of two faces of the epicuticle: a inner face containing numerous particles, and a almost smooth outer face. Deep etching replicas confirmed the existance of these two faces of the epicuticle showing in some replicas two particle populations on the outer face of L4 and adult forms of C. elegans. Also a previously unrecognized structure was noted in the cuticle of C. elegans, a matrix composed by network of globular and filamentous structures, leaving in between them spaces, which probably are occupied by water in the living adult and L4 larvae specimen. This network demonstrates either a compact nature or loose nature according to their cuticle location. Deep etching replicas of the adults nematode revealed large spaces between the cortical and basal layers which are regularly interrupted by struts connecting each other by fibers in a particular arrangement. PMID- 7491626 TI - Osteoblasts from the toothless (osteopetrotic) mutation in the rat are unable to direct bone resorption by normal osteoclasts in response to 1,25-dihydroxyvitamin D. AB - Osteopetrosis describes a diversified group of metabolic bone disorders characterized by a generalized, skeletal sclerosis resulting from reduced osteoclast-mediated bone resorption. The toothless (tl) osteopetrotic mutation in the rat is characterized by few osteoclasts and the inability to be cured by transplants of hemopoietic stem cells. This implies that the defect(s) responsible for reduced osteoclast activity in tl rats is within the skeletal microenvironment (cells or matrices). Osteoblasts and their products are known to play a role in regulating bone resorption and abnormalities in the osteoblast population in tl rats have been reported. The purpose of this study was to determine whether osteoblasts isolated from tl mutant rats, when cultured with normal osteoclasts, could increase bone resorption (pit formation) in response to stimulation by 1,25 dihydroxyvitamin D (1,25(OH)2D). The addition of 1,25(OH)2D produced a highly significant response in normal osteoblast cocultures but no response in mutant cultures. A dose response study with 1,25(OH)2D (10(-6) to 10( 9)M) revealed that mutant osteoblasts are unable to increase osteoclast activity. These data indicate that the vitamin D receptor-signal transduction pathway in tl rats needs to be examined. PMID- 7491624 TI - Type II cytokeratin expression in adult vertebrate spinal cord. AB - The phylogenetic evolution of the expression of type II cytokeratins (CKs) in the spinal cord of different adult vertebrates has been studied using an anti-CK immunohistochemical technique. Type II CK expression was stronger in lower vertebrates, specially anuran amphibians, than in higher vertebrates. No CK expression was found either in reptiles or birds, but a weak expression was demonstrated in mammals. The main neuroectodermal cell implicated in CK expression was the ependymocyte; some CK-positive radial astrocytes were also found in amphibians and fish, but neither CK-positive astrocytes nor neurons were observed in any vertebrate group. The functional significance of CK expression in the vertebrate spinal cord is not known. CKs do not have a consistent pattern of expression amongst vertebrates; however, the most common site is the ependyma. PMID- 7491627 TI - Effect of long-term insulin exposure on insulin binding in Tetrahymena pyriformis. AB - Previous studies have indicated that Tetrahymena pyriformis can bind the vertebrate hormone insulin. Conventional microscopic studies were conducted to determine the effect of acute and long-term (LTE) insulin exposure on insulin binding. Stock cultures included cells never exposed to insulin and cultures grown in medium containing 6 mg/ml insulin. Logarithmic cultures were exposed to porcine insulin concentrations of 0 and 6 mg/ml for 1 h (insulin treated, (IT)) after 0, 48 h, 1, 3, or 6 months of LTE insulin exposure. 24 h after the 1 h insulin treatment, the cells were fixed, exposed to porcine insulin (antigen), processed immunocytochemically using a primary antibody to porcine insulin and a secondary antibody immunocytochemistry kit, and examined for staining intensity by video image analysis. Morphological observations confirm that T. pyriformis does bind insulin whether or not the cells have had prior exposure to insulin. IT increases insulin binding (up-regulation) in previously unexposed cells (control, P < 0.01) and produces a further amplification in cells having prior acute exposure (48 h) to insulin (P < 0.01). However, LTE exposure to insulin (1, 3 and 6 months) caused a decrease in insulin binding (down-regulation) after IT (P < 0.01) such that LTE-IT cells were not different from control cells following 1, 3 or 6 months of chronic insulin exposure to insulin. Staining intensity was not different between IT cells and cells cultured with insulin throughout the six month study. Results suggest that insulin binding sites of T. pyriformis are subject to regulatory processes similar to those of metazoans. PMID- 7491628 TI - Gap junctional communication in primary culture of cells derived from human aortic intima. AB - Intercellular communication via gap junctions plays an important role in the regulation and homeostasis. The presence of gap junctions and the efficiency of their function directly correlates with the degree of cell differentiation in a tissue. In the present study, gap junctional communication has been investigated in a primary culture of highly differentiated mesenchymal cells (subendothelial smooth muscle cells isolated from grossly normal and atherosclerotic areas of human aorta) and in poorly differentiated cells of mesenchymal origin (adult human skin fibroblasts as well as skin fibroblasts and aortic smooth muscle cells, derived from human fetus). The fluorescent dye transfer technique was used in this study. In cell cultures isolated from grossly normal and atherosclerotic aorta, the number of cells coupled via gap junctions increased with cell density and reached a plateau at a cell density of 50 to 70 cells/mm2. In cultures of normal aortic cells the number of coupled cells was 23.0 +/- 4.1 per injected cell and was significantly higher than in cultures of atherosclerotic cells (16.4 +/- 2.1, p < 0.05). Gap junctional communication between cells loaded with lipid inclusions was 2.4-fold lower than between cells free of excess intracellular lipids. In cultures of human skin fibroblasts the rate of intercellular communication was lower than in cultures of normal aortic cells and was comparable to that in cultures of atherosclerotic cells. There was practically no cell-to-cell communication in cultures of fetal cells. It is hypothesized that the reduced gap junctional communication in atherosclerotic human aorta is associated with the alterations in the degree of smooth muscle cell differentiation and impairs the function of the intima in atherosclerosis. PMID- 7491629 TI - Gasoline as an oncological problem. PMID- 7491631 TI - Toxicity and carcinogenicity of t-butyl alcohol in rats and mice following chronic exposure in drinking water. AB - t-Butyl alcohol (TBA) was administered in drinking water to F344/N rats and B6C3F1 mice for two years using 60 animals/dose/sex/species. Male rats received doses of 0, 1.25, 2.5, or 5 mg/ml and females received 0, 2.5, 5, or 10 mg/ml, resulting in average daily doses of approximately 85, 195, or 420 mg TBA/kg body weight for males and 175, 330, or 650 mg/kg for females. Ten rats per group were evaluated after 15 months. Male and female mice received doses of 0, 5, 10, or 20 mg/ml, resulting in average daily doses of approximately 535, 1,035, or 2,065 mg TBA/kg body weight for males and 510, 1,015, or 2,105 mg/kg for females. Survival was significantly reduced in male rats receiving 5 mg/ml, female rats receiving 10 mg/ml, and male mice receiving 20 mg/ml. Long-term exposure to TBA produced increased incidences of renal tubule adenoma and carcinoma in male rats; transitional epithelial hyperplasia of the kidney in male and female rats; follicular cell adenoma of the thyroid in female mice; and follicular cell hyperplasia of the thyroid and inflammation and hyperplasia of the urinary bladder in male and female mice. In addition, a slight increase in follicular cell adenoma or carcinoma of the thyroid (combined) in male mice may have been related to the administration of TBA. PMID- 7491630 TI - Methyl-tertiary-butyl ether (MTBE)--a gasoline additive--causes testicular and lymphohaematopoietic cancers in rats. AB - In the framework of a series of experiments conducted to evaluate the carcinogenic effects of oxygenated gasoline additives, MTBE was analyzed in an oral lifetime carcinogenicity study using 8-week-old male and female Sprague Dawley rats. These experiments were part of a large research project on gasoline carcinogenicity performed at the Bentivoglio (BT) Castle Cancer Research Center of the Ramazzini Foundation and of the Bologna Institute of Oncology, MTBE, dissolved in oil, was administered by stomach tube at the doses of 1000, 250, or 0 mg/kg b.w., once daily, four days weekly, for 104 weeks. The animals were maintained until natural death. The last animal died 166 weeks after the start of the experiment, i.e., at 174 weeks of age. Under the tested experimental conditions, MTBE was shown to cause an increase in Leydig interstitial cell tumors of the testes and a dose-related increase in lymphomas and leukemias in female rats. PMID- 7491632 TI - Methyl tertiary butyl ether (MTBE)--evaluation of MTBE carcinogenicity studies. PMID- 7491633 TI - Developmental toxicity study of piperonyl butoxide in CD rats. AB - Piperonyl butoxide was administered to pregnant rats by gavage at a level of 0 (control), 630, 1065, and 1800 mg/kg bw on days 11-12 of gestation. The animals were killed on day 20 of gestation. Average maternal body weight gain (gestational days 11-20) was significantly reduced in the 1065 and 1800 mg/kg bw groups. Total resorption rate was significantly increased in the 1800 mg/kg bw group and those effects were significantly dose-related. The average fetal body weight of each sex was significantly reduced in the 1065 and 1800 mg/kg bw groups. External limb deformity (oligodactyly, syndactyly, and polydactyly) was significantly increased in the 1065 and 1800 mg/kg bw groups in a dose-related manner. The dose levels of piperonyl butoxide in the present study produced limb deformities in rats. PMID- 7491634 TI - Hydrogen sulfide and reduced-sulfur gases adversely affect neurophysiological functions. AB - Hydrogen sulfide (H2S) above 50 parts per million (ppm) causes unconsciousness and death. Lower doses of H2S and related gases have been regarded as innocuous, but the effects of prolonged exposure have not been studied. This study was designed to determine whether people exposed to sulfide gases as a result of working at or living downwind from the processing of "sour" crude oil demonstrate persistent neurobehavioral dysfunction. Thirteen former workers and 22 neighbors of a refinery complained of headaches, nausea, vomiting, depression, personality changes, nosebleeds, and breathing difficulties. Their neurobehavioral functions and a profile of mood states (POMS) were compared to 32 controls, matched for age and educational level. The exposed subjects' mean values were statistically significantly abnormal compared to controls for two-choice reaction time, balance (as speed of sway), color discrimination, digit symbol, trail-making A and B, and immediate recall of a story. Their POMS scores were much higher than those of controls. Visual recall was significantly impaired in neighbors, but not in exworkers. It was concluded that neurophysiological abnormalities were associated with exposure to reduced sulfur gases, including H2S from crude oil desulfurization. PMID- 7491635 TI - General toxicity and reproductive screen of liquid propellant XM46 administered in the drinking water of Sprague-Dawley rats. AB - Liquid propellant XM46 is being considered as a replacement for solid propellants, both as part of a regenerative injection gun system and as a working fluid in an electrothermal gun system. The XM46 formulation contains hydroxylammonium nitrate, triethanolammonium nitrate, and water. Male and female Sprague-Dawley rats received XM46 in drinking water containing 2.0, 1.0, 0.2, or 0.0 g XM46/liter throughout a 90-day study. Mating occurred following 14 days of treatment. One-half the male rats per group were necropsied after 28 days of treatment; the remaining males and all dams were necropsied following 90 days of treatment. No mortality occurred in any of the parental animals during the study. The study did not demonstrate any adverse effects on reproduction or litter parameters. Hemolytic anemia and methemoglobinemia were common in both sexes of rats. Splenomegaly was found in both sexes; in male rats as early as 28 days. Exposures via drinking water containing XM46 for 90 days did not result in any decrease in reproductive performance in male or female rats, but it did result in clinical signs of hemolytic anemia at doses as low as 17 mg/kg/day. PMID- 7491636 TI - Environmental personal injury litigation as one source of response effects: findings from the National Exposure Registry. AB - The potential for error in survey responses obtained from people involved with environmental personal injury litigation was examined in a registry of persons exposed to the chemical trichloroethylene. Two subgroups were selected and compared: environmental personal injury plaintiffs and nonlitigants residing in the same community. Self-reported information on demographic characteristics revealed no statistically significant differences. Although plaintiffs reported higher rates of symptoms and health problems, only 2 of the 20 comparisons on health were statistically significant. The overall similarity between the two groups suggests that environmental personal injury plaintiffs may be no more likely than nonlitigants to provide inaccurate information in health surveys. PMID- 7491637 TI - The National Exposure Registry: procedures for establishing a registry of persons environmentally exposed to hazardous substances. AB - The Agency for Toxic Substances and Disease Registry has, as mandated in Superfund legislation, established the National Exposure Registry (NER). The purpose of the NER is to assess and evaluate the potential relationship between adverse health effects and environmental exposure for an exposed population, particularly the relationship between chronic health effects and long-term, low level chemical exposures. The NER's primary goal is to facilitate epidemiology research by establishing multiple data bases (subregistries) that contain demographic, environmental, and health information on large populations exposed to selected chemicals. The Registry data mainly serve the purpose of being hypothesis-generating rather than hypothesis-testing. The NER is currently composed of subregistries of: (1) persons exposed to volatile organic compounds (VOCs)--a subset of registrants in whom trichloroethylene (TCE) is the primary VOC exposure, but others are present (N = 4,832), a subset in whom benzene is the primary VOC exposure (N = 1,142), and a subset in whom trichloroethane (TCA) and TCE are the highest VOC exposures (N = 3,666); and (2) persons with dioxin exposure (N = 250). Chromium and radioactive substances subregistries are planned. PMID- 7491638 TI - Acute stroke treatment trials in the United States. Rethinking strategies for success. PMID- 7491639 TI - Treatment of acute ischemic stroke. Challenging the concept of a rigid and universal time window. PMID- 7491640 TI - Stroke in the People's Republic of China. I. Geographic variations in incidence and risk factors. AB - BACKGROUND AND PURPOSE: Stroke is a major cause of death in the People's Republic of China (PRC), and the geographic distribution of stroke death varies substantially. We conducted an ecological analysis to investigate the relationship of prevalence of hypertension and other risk factors with stroke incidence and mortality in the PRC. METHODS: The relationship between prevalence of risk factors and stroke incidence and mortality in the PRC was assessed in 29 provinces by examining risk factor data from PRC national surveys and incidence and mortality from the 1986 PRC National Stroke Study. RESULTS: A highly statistically significant correlation between prevalence of hypertension and stroke incidence (r = .838, P < .001) and mortality (r = .841, P < .001) was observed. Prevalence of hypertension explained over 70% of the geographic variability in stroke incidence (partial R2 = .703) and mortality (partial R2 = .707) in the PRC. There was a north-south gradient, with a higher prevalence of hypertension and stroke incidence and mortality in the north compared with the south of the country. In multiple Poisson regression analysis, a 10% increase in the prevalence of hypertension was associated with a 2.80-fold higher incidence and 2.68-fold higher mortality from stroke, whereas a 10% increase in the prevalence of alcohol consumption was associated with a 29% higher incidence and a 16% higher mortality from stroke. A 10% increase in the prevalence of cigarette smoking was associated with a 19% higher mortality from stroke. CONCLUSIONS: In the PRC, the prevalence of hypertension is strongly associated with the risk of stroke, and the geographic variation in stroke incidence and mortality is due mainly to differences in the prevalence of hypertension. PMID- 7491641 TI - Stroke in the People's Republic of China. II. Meta-analysis of hypertension and risk of stroke. AB - BACKGROUND AND PURPOSE: In the People's Republic of China (PRC), stroke accounts for over 1 million deaths per year, more than three times the number of deaths from ischemic heart disease. To quantify the effect of hypertension on stroke in the PRC, we conducted a meta-analysis and estimated the population attributable risk of stroke due to hypertension. METHODS: We searched the Chinese Medical Literature Index and MEDLINE databases to identify studies on the relationship between hypertension and stroke conducted in the PRC. The overall relative risk of stroke in hypertensive persons and the population attributable risk of stroke due to hypertension were calculated by pooling the risk estimates from individual studies. RESULTS: A total of 12 epidemiological studies with 2379 stroke cases was identified. The overall relative risk of stroke associated with hypertension was 5.43 (95% confidence interval [CI], 4.62 to 6.39). The relative risk was similar for hemorrhagic (5.44; 95% CI, 3.99 to 7.40) and ischemic (5.25; 95% CI, 3.95 to 6.98) stroke. On the basis of data from the 1990 PRC population census, vital statistics, and the 1991 PRC National Hypertension Survey, the excess annual mortality from stroke due to hypertension was estimated to be at least 545,000 deaths, with a population attributable risk of 49.5%. CONCLUSIONS: Our data suggest that hypertension is an important risk factor for stroke in the PRC and that hypertension may account for about half of all deaths from stroke in that country. Treatment and prevention of hypertension should be an important societal goal in the PRC. PMID- 7491642 TI - Delay in neurological attention and stroke outcome. Cerebrovascular Diseases Study Group of the Spanish Society of Neurology. AB - BACKGROUND AND PURPOSE: Despite efforts to reduce the delay between stroke onset and new interventional treatments, no studies have analyzed the repercussions of early neurological attention on the clinical outcome of stroke patients. METHODS: Data were obtained from 721 patients admitted consecutively for a transient ischemic attack or stroke to the neurology departments of 18 Spanish hospitals that followed the same diagnostic and therapeutic guidelines in the acute phase. Factors assessed were age, sex, Canadian Stroke Scale score on admission, previous Barthel Index, and delay before attention by the first physician, by emergency services, by a neurologist, and before hospitalization. Patients' outcomes were classified as good (Barthel Index > 60) or poor (Barthel Index < or = 60 or in-hospital death) depending on patient's functional capacity on discharge. The individual contribution of each of these variables on clinical outcome was estimated with logistic regression analysis. RESULTS: Patients in worse neurological condition on admission presented earlier to the first physician, emergency department, and neurologist. The mortality rate was not significantly modified by early or late presentation at the different medical stages. Logistic regression analysis revealed that the relative risk of poor outcome in patients seen by the neurologist after the first 6 hours from symptom onset was 5.6 (95% confidence interval, 3.4 to 9.2) (P < .0001). Multiple linear regression analysis showed that the delay before the patient received neurological attention correlated positively with the duration of hospitalization (P < .0001). The delays before the patient was seen by the first physician or the emergency department and before hospitalization were not independently related to clinical outcome or length of hospitalization. CONCLUSIONS: Early neurological attention in acute stroke is related to better functional outcome and shorter hospitalization. PMID- 7491643 TI - Emergency physicians. Accuracy in the diagnosis of stroke. AB - BACKGROUND AND PURPOSE: The role of emergency physicians in trials of acute stroke therapy is expanding. We investigated the ability of emergency physicians to accurately identify patients with stroke. METHODS: We reviewed all 446 patients who were evaluated in the emergency department and had an inpatient admitting or discharge diagnosis of ischemic or hemorrhagic stroke from May 1, 1992, to June 30, 1993. The final admitting diagnosis made by the emergency physician was compared with the final hospital discharge diagnosis, 95% of which were made by neurologists or neurosurgeons. RESULTS: All 76 patients with a final discharge diagnosis of intracerebral or subarachnoid hemorrhage were correctly diagnosed by the emergency physicians (sensitivity, 100%; positive predictive value, 100%). Of the 351 patients with a final discharge diagnosis of ischemic stroke or transient ischemic attack, 346 were correctly identified by the emergency physicians (sensitivity, 98.6%; positive predictive value, 94.8%). Nineteen patients were diagnosed with stroke or transient ischemic attack by the emergency physician but had a final discharge diagnosis other than stroke. Discharge diagnoses included paresthesia or numbness of unknown causes (3), seizure (2), complicated migraine (2), peripheral neuropathy (2), cranial nerve neuropathy (2), psychogenic paralysis (1), and other (7). CONCLUSIONS: Emergency physicians at a large urban teaching hospital with a comprehensive stroke intervention program can accurately identify patients with stroke, particularly hemorrhagic stroke. If similar accuracy can be documented in other types of hospitals, emergency physicians may become key providers of urgent stroke intervention. PMID- 7491644 TI - Symptom control, communication with health professionals, and hospital care of stroke patients in the last year of life as reported by surviving family, friends, and officials. AB - BACKGROUND AND PURPOSE: The needs of and appropriate service provision for patients dying from stroke have received little attention. The quality of care in the last year of life received by a population-based sample of stroke patients is described here, focusing on symptom control, communication with health professionals, and hospital care. METHODS: Secondary analysis was made of data from the Regional Study of Care for the Dying, a retrospective interview survey in 20 nationally representative English health districts. Subjects were 237 persons who died from stroke in 1990. Of informants, 20% were spouses, 48% relatives, 11% friends or neighbors, and 20% officials. RESULTS: More than half the patients were reported to have experienced pain (65%), mental confusion (51%), low mood (57%), and urinary incontinence (56%) in the last year of life. Pain control was inadequate: 51% of those treated for pain by hospital doctors and 45% of those treated by general practitioners were reported to have received treatment that relieved pain partially if at all. One third of respondents thought that hospital doctors had been too rushed (37%), and 25% thought that the patient had had insufficient choice about treatment. Two fifths had been unable to get all the information they had wanted about the patient's condition. CONCLUSIONS: Improvements in symptom control and psychosocial support for patients who die from stroke are needed, as is better communication between health professionals and patients and their families. Education of doctors and nurses working with stroke patients in the principles of palliative care may help ensure that all dying stroke patients receive high-quality care. PMID- 7491645 TI - Neurological and functional outcome in patients with supratentorial hemorrhages. A prospective study. AB - BACKGROUND AND PURPOSE: A prospective study was performed to evaluate neurological and functional outcome after spontaneous supratentorial bleeding. The aim of the study was to determine whether clinical or neuroradiological parameters could predict the outcome of these patients during the first hours of hospitalization. METHODS: Two hundred seventy-nine patients--52 with thalamic, 87 with putaminal, and 140 with lobar hemorrhages--were followed prospectively and examined on admission and at 2 weeks, 3 months, and 6 months after onset. The patients underwent clinical (according to the Glasgow Coma Scale) and neuroradiological examinations on admission and were scored clinically and functionally (according to Stroke Severity score and Barthel Index) on the follow up periods. Risk factors and the correlation between findings on admission and the latest clinical and functional results were calculated with the chi 2 test, Pearson correlation test, and Student's t test. Multivariate analysis was calculated with the stepwise regression test. RESULTS: In all of the bleeding locations, lethal outcome was significantly correlated with size of the hematoma (P < .001) and Glasgow Coma Scale score on admission (P < .001). Intraventricular blood expansion was found to have a better prognosis in thalamic bleeding (P < .007) and a worse prognosis in lobar hemorrhage (P < .01). The functional outcome after 6 months was directly correlated with the size of the bleeding area in lobar and putaminal hemorrhages. No correlation was found in thalamic bleeding. A worse functional outcome was found in putaminocapsular bleeding (P = .004) and in patients with ischemic heart disease. A limited better recovery prognosis was found in patients with lobar hematoma in the temporal lobe (P = .052). CONCLUSIONS: The probability of lethal outcome can be calculated on admission in all patients with supratentorial bleeding and in correlation with the location and size of the bleeding area and level of consciousness. Intraventricular expansion of blood is a better prognostic factor in thalamic bleeding and a worse one in lobar hematoma. Functional outcome is correlated with size of the bleeding area and level of consciousness on admission in putaminal and lobar hemorrhages but has no correlation to thalamic hemorrhage. PMID- 7491646 TI - Amphetamine paired with physical therapy accelerates motor recovery after stroke. Further evidence. AB - BACKGROUND AND PURPOSE: In animal models of brain injury, administration of numerous pharmaceuticals is reported to facilitate functional recovery. However, only drugs that increase the release of norepinephrine have been shown to promote recovery when administered late (days to weeks) after injury. To determine whether these findings were applicable to humans, we administered the norepinephrine stimulant dextroamphetamine, paired with physical therapy, to hemiplegic stroke patients. METHODS: Ten hemiplegic patients who suffered an acute ischemic infarction were entered between days 16 and 30 after onset and randomly assigned to receive either 10 mg of dextroamphetamine or a placebo orally every fourth day for 10 sessions paired with physical therapy. The Fugl Meyer Motor Scale was used at baseline, within each session, and for 12 months after onset as the dependent measure. Confounding medications such as alpha adrenergic antagonists or agonists were excluded in all subjects. RESULTS: Although there were no differences between the groups at baseline (P = .599), there was a significant (P = .047) difference between the groups when the drug had been discontinued for 1 week and at the 12-month follow-up visit (P = .047). CONCLUSIONS: Administration of dextroamphetamine paired with physical therapy increased the rate and extent of motor recovery in a small group of hemiplegic stroke patients. These data support and extend previous findings of the facilitatory aspects of certain types of drugs on recovery from brain injury. The use of neuromodulation may allow the nervous system to adapt previously unused or alternative pathways to relevant external input. PMID- 7491647 TI - Safety of hypertensive hypervolemic therapy with phenylephrine in the treatment of delayed ischemic deficits after subarachnoid hemorrhage. AB - BACKGROUND AND PURPOSE: Hypertensive hypervolemic therapy has been shown to reverse delayed ischemic deficits after aneurysmal subarachnoid hemorrhage. Concern has been raised about systemic complications of therapy, including pulmonary edema and myocardial ischemia, especially when high doses of vasopressors are used. Patients in whom delayed ischemic deficits were treated with hypervolemia and phenylephrine were prospectively evaluated for signs of systemic toxicity. METHODS: Twenty-four consecutive patients treated with hypertensive hypervolemic therapy after aneurysmal subarachnoid hemorrhage were studied. Sixty-seven percent had underlying cardiac disease, vascular disease, or hypertension. No patient was excluded because of age or preexisting cardiac disease. Patients were closely monitored for signs of congestive heart failure (physical examination, chest x-ray films, arterial blood gases, cardiac index, pulmonary artery wedge pressure, and oxygen requirement). Indicators of cardiac ischemia and other extracerebral toxicity that were monitored included cardiac enzymes, electrocardiograms, serum creatinine, electrolyte and lactic acid levels, gastrointestinal motility, and urine output. RESULTS: Volume expansion and phenylephrine infusion produced an increase in several hemodynamic parameters including pulmonary artery wedge pressure, which rose 28% (13 +/- 3.6 to 16 +/- 1.9 mm Hg), mean arterial blood pressure, which rose 25% (99 +/- 12.5 to 123 +/- 11.4 mm Hg), and systemic vascular resistance, which rose 46% (1234 +/- 294 to 1739 +/- 315 dyne.s-1.cm-5); however, there was no change in cardiac index (3.9 +/- 0.9 to 4.0 +/- 0.6 L.min-1.m-2). There were no clinically significant episodes of pulmonary edema requiring a change in vasopressor therapy and no myocardial infarctions. Phenylephrine was stopped in only one patient (incidence, 4%; 95% confidence interval, 0% to 12%), who developed an exacerbation of his preexisting bradycardia. There was no evidence of noncardiac organ system toxicity. Eighty-eight percent of the patients exhibited neurological improvement. CONCLUSIONS: Hypertensive hypervolemic therapy with the use of high dose phenylephrine can be administered with acceptable systemic toxicity, even in patients with previous cardiac disease, provided that close monitoring is performed. To minimize risk, aggressive treatment should probably be reserved for patients with signs of delayed ischemia rather than administered prophylactically. PMID- 7491648 TI - Central benzodiazepine receptor distribution after subcortical hemorrhage evaluated by means of [123I]iomazenil and SPECT. AB - BACKGROUND AND PURPOSE: [123I]Iomazenil is a single-photon emission computed tomography (SPECT) tracer that selectively binds to central benzodiazepine receptor in the neuron membrane. With this ligand, we studied the central benzodiazepine receptor distribution in the cortex remote from subcortical hematoma in intracerebral hemorrhage patients. METHODS: Four patients with unilateral putaminal hemorrhage and one patient with right thalamic hemorrhage were studied (mean +/- 1 SD age, 50.0 +/- 8.8 years). The initial volume of hematoma ranged from 4.3 to 31.0 mL (mean +/- 1 SD, 17.5 +/- 12.3 mL). SPECT images obtained 3 hours after intravenous administration of [123I]iomazenil (167 MBq/750 ng) were analyzed. In three patients, perfusion was evaluated with [123I]IMP. On SPECT images, the radioactivity ratio of the ipsilateral to the contralateral cerebral cortex (I/C ratio) or of the contralateral to the ipsilateral cerebellar hemisphere (C/I ratio) was measured. RESULTS: The I/C ratio for iomazenil was significantly decreased compared with unity in the temporal lobe (0.84 +/- 0.08, P < .01) and the parietal lobe (0.87 +/- 0.10, P < .05), but the C/I ratio in the cerebellum (1.00 +/- 0.03) was not. The C/I ratio for perfusion in the cerebellar cortex (0.83 +/- 0.04, P < .01) was significantly decreased compared with that in normal subjects. CONCLUSIONS: Central benzodiazepine receptor-[123I]iomazenil binding was decreased in the ipsilateral cerebral cortex remote from the subcortical hematoma. This preliminary result may facilitate further study of the potential damage of the cortical neurons remote from subcortical hematoma. PMID- 7491649 TI - Measurement of infarct size using MRI predicts prognosis in middle cerebral artery infarction. AB - BACKGROUND AND PURPOSE: An accurate measure of the severity of ischemic insult and the resulting prognosis is needed to assess the effectiveness of new treatments for acute stroke. We studied the reproducibility and accuracy of measurements of infarct volume with MRI and correlated the measurements with outcome. METHODS: Infarct volume was measured on T2-weighted images with the Analyze image analysis software. This technique was found to be highly accurate and reproducible. RESULTS: Measurements of infarct volume were found to be highly accurate and reproducible. Twenty-one patients (mean age, 66.5 years; range, 28 to 90 years) with cortical middle cerebral artery territory infarcts in whom adequate data could be obtained were studied within 72 hours from onset (mean delay to MRI, 27.5 hours; range, 5 to 72 hours). The Scandinavian Stroke Scale was used to calculate a prognostic score, and clinical outcome was assessed at 3 months. Infarct volume was found to significantly predict outcome. Mean infarct volume in the independent patients was 35.7 +/- 29.7 cm3 compared with 88.3 +/- 71.3 cm3 in dependent patients and 166.5 +/- 65.9 cm3 in dead patients (F = 10.52, P < .001). Patients with an initial infarct volume less than 80 cm3 were found to have a better outcome than those with larger infarct volumes. Secondary hemorrhage visible on MRI also predicted a poor outcome. In contrast, the Scandinavian Stroke Scale did not significantly predict outcome. CONCLUSIONS: The results demonstrate that measurement of the size of middle cerebral artery infarction with MRI is a useful tool in assessing prognosis and will have a valuable role in assessing new therapeutic agents. PMID- 7491650 TI - Prediction of reflex sympathetic dystrophy in hemiplegic patients by electromyographic study. AB - BACKGROUND AND PURPOSE: This study was designed to investigate the correlation between reflex sympathetic dystrophy syndrome (RSDS) in hemiplegic patients and spontaneous electromyographic (EMG) activity, as well as to determine the predictive value of spontaneous EMG activity in early diagnosis of RSDS. METHODS: An EMG and nerve conduction velocity study of the weak upper limb was conducted on 70 hemiplegic patients at 3 to 4 weeks after cerebrovascular disease (either cerebral hemorrhage or infarction). Clinical assessment for development of the RSDS was done during the following 6 months. The correlation of RSDS development with the presence of spontaneous EMG activity and certain clinical parameters (including sex, age, side affected, cause of stroke, sensory impairment, spasticity, and shoulder subluxation) was analyzed statistically. RESULTS: Of the 46 patients who exhibited spontaneous activity, 30 (65%) developed clinical RSDS in their hemiplegic upper extremity, whereas only 1 (4%) of the other 24 patients with no spontaneous EMG activity developed clinical RSDS within 6 months after the onset of hemiplegia (P < .001). The correlation of RSDS development with the presence of shoulder subluxation and sensory impairment in the hemiplegic side was statistically significant. Neither age, sex, severity of spasticity, nor etiology of stroke had a significant correlation with the development of clinical RSDS. CONCLUSIONS: There is significant correlation between the presence of spontaneous EMG activity and the development of clinical RSDS in the hemiplegic upper extremity after stroke. It is concluded that spontaneous EMG activity in the hemiplegic hands of stroke patients might be a good predictor of the future development of clinical RSDS. PMID- 7491651 TI - Interpretation of embolic phenomena during carotid endarterectomy. AB - BACKGROUND AND PURPOSE: Air and particulate emboli are a major source of morbidity during carotid endarterectomy (CEA); however, amplitude overload and poor time resolution have restricted the ability of transcranial Doppler ultrasound to differentiate between the two. METHODS: We have now overcome these two limitations by (1) rerouting embolic signals away from the audio frequency amplifier to avoid amplitude overload and (2) substituting the Wigner distribution function for the fast Fourier transform to improve time and frequency resolution. Thus, we can now accurately determine embolic duration and embolic velocity, the product of which is the sample volume length (SVL). This measurement represents the physical distance over which an embolic signal can be detected. The underlying hypothesis was that air reflected more ultrasound and would therefore be detected over a greater SVL. RESULTS: The median SVL (interquartile range) for 75 in vitro air emboli was 1.97 cm (range, 1.70 to 2.35) compared with 0.27 cm (range, 0.16 to 0.43) for 185 particulate emboli detected during the dissection phase of CEA. Off-line analysis on an additional 560 embolic signals detected during different phases of CEA suggested that 46 of 143 (32%) of emboli immediately after shunt insertion were particulate, as were 19 of 33 (58%) occurring during shunting, 28 of 78 (36%) after restoration of flow in the external carotid artery, 23 of 251 (9%) after restoration of flow in the internal carotid artery, and 55 of 55 (100%) of those emboli detected during the early recovery phase. CONCLUSIONS: This development provides objective physical criteria upon which embolus characterization (particulate/air) can be based. This could have major implications for future patient monitoring with respect to modification of surgical technique and pharmacological intervention. PMID- 7491652 TI - Can cerebrovascular reactivity be measured with near-infrared spectroscopy? AB - BACKGROUND AND PURPOSE: We used near-infrared spectroscopy (NIRS) to monitor the cerebral oxygenation changes during CO2 reactivity tests. METHODS: Fifty healthy volunteers were examined (age range, 19 to 68 years). The monitored parameters were as follows: transcranial Doppler (TCD) time-averaged middle cerebral artery flow velocity end-tidal CO2 (EtCO2); change in concentration of cerebral oxyhemoglobin (HbO2), deoxyhemoglobin (Hb), and total hemoglobin; mean arterial blood pressure; peripheral arterial oxygen saturation (SaO2); and extracranial tissue perfusion with the use of cutaneous laser-Doppler flowmetry. The examination protocol included both hypercapnia and hypocapnia. The cerebrovascular reactivity indexes were calculated as follows: TCD, relative change in flow velocity per 1 kPa increase in EtCO2; NIRS, absolute change in HbO2, Hb, and total hemoglobin concentration (micromoles per liter) per 1 kPa increase in EtCO2. RESULTS: Mean middle cerebral artery flow velocity was found to be 58 cm/s at a mean baseline EtCO2 of 4.7 kPa. Mean cerebrovascular reactivities were as follows: TCD, 24%/kPa (SEM, 1.1); HbO2, 2.06 mumol/L per kilopascal (SEM, 0.08); Hb, -0.63 mumol/L per kilopascal (SEM, 0.09); and total hemoglobin concentration, 1.44 mumol/L per kilopascal (SEM, 0.1). Statistical analysis revealed significant correlation between reactivities calculated with the use of NIRS and TCD (P < .001). Although some fluctuations were observed in SaO2 and laser-Doppler flux, they were not correlated with either EtCO2 or NIRS. CONCLUSIONS: NIRS signal changes in HbO2, Hb, and total hemoglobin concentration are very sensitive to alterations in EtCO2, which are largely independent of extracranial tissue perfusion. NIRS may be developed as an alternative method for testing cerebrovascular reactivity and may be of particular clinical importance when the ultrasound window is poor. PMID- 7491653 TI - Application of transcranial Doppler sonography to evaluate cerebral hemodynamics in carotid artery disease. Comparative analysis of different hemodynamic variables. AB - BACKGROUND AND PURPOSE: Transcranial Doppler sonography in combination with manipulation of cerebral resistance vessels is widely used to screen patients with suspected intracranial hemodynamic disturbances. Maximal flow velocity (Vmax), mean flow velocity (Vmean), cerebral pulsatility index (CPi), and cerebral resistance index (CRi) have all been used to describe cerebral hemodynamics. The present study examined CO2 reactivity of the above hemodynamic variables with respect to its variability between different age groups and its capability to discriminate between normal and abnormal findings. METHODS: Absolute and relative CO2 reactivity of Vmax, Vmean, CRi, and CPi were determined in both hemispheres in 30 young and 37 elderly control subjects and in 245 consecutive patients with strictly unilateral symptomatic (n = 101) or asymptomatic (n = 144) carotid artery disease (> 80% stenosis or occlusion). RESULTS: Hemispheric reactivities of Vmean, CRi, and CPi were significantly age dependent. Hemispheric Vmax reactivity and interhemispheric differences of individual reactivities (except absolute CPi reactivity) did not vary with age and could therefore be used to define normal values. Patient classification according to these values revealed different frequencies of subjects with pathological findings (3% for hemispheric Vmax reactivity, 5% to 7% for interhemispheric differences of Vmax or Vmean reactivity, 39% and 45% for interhemispheric differences of relative CRi and CPi reactivity, respectively). CONCLUSIONS: Hemispheric reactivities are less suitable to evaluate cerebral hemodynamics than interhemispheric differences, since most of the latter do not vary with age. However, interhemispheric differences vary with respect to their discriminatory power. Power is low for interhemispheric differences of Vmax and Vmean reactivity, since the corresponding frequencies of abnormal findings do not differ from the 5% frequency expected in the reference population (reference range defined as mean +/- 2 SD). With respect to the discriminatory power, interhemispheric differences of relative CRi and CPi reactivity may be superior to other parameters. PMID- 7491654 TI - Transcranial color-coded duplex sonography in cerebral arteriovenous malformations. AB - BACKGROUND AND PURPOSE: It is well known that significant changes in cerebral hemodynamics may occur during the treatment of cerebral arteriovenous malformations with the complication of intracerebral hemorrhage and parenchymal edema. We used transcranial color-coded duplex sonography to study alterations in blood flow velocities during staged embolization. METHODS: Forty-one patients aged 40 +/- 13 years (mean +/- SD) with angiographically proven cerebral arteriovenous malformations were studied. The blood flow velocities of the anterior, middle, and posterior cerebral arteries were measured in 16 patients with supratentorial arteriovenous malformations, both before the first and then after each successive embolization (three to seven treatments). RESULTS: In 29 of 41 patients (71%), transcranial color-coded duplex sonography satisfactorily revealed the malformations and their main feeders. After the final embolization, we found a reduction in the peak flow velocity in treated feeders of 23 +/- 28% compared with the values before the first embolization. The untreated feeders showed an increase in peak flow velocities of 12 +/- 23% as an expression of increased collateral flow. After the treatment of the supplying feeders, we observed a reduction in flow velocity of 25 +/- 13% in seven patients, with cross filling of the arteriovenous malformation through the contralateral anterior cerebral artery and the anterior communicating artery. CONCLUSIONS: The technical advantage of transcranial color-coded duplex sonography compared with transcranial Doppler sonography is that it allows the exact identification of different feeding arteries in arteriovenous malformations. Repeated measurements during stepwise embolization with corrected insonation angle are easily achieved, and noninvasive quantification of hemodynamic changes is possible. The method may be helpful in the planning of the different steps of embolization. PMID- 7491655 TI - Cerebral vasoreactivity assessed with transcranial Doppler and regional cerebral blood flow measurements. Dose, serum concentration, and time course of the response to acetazolamide. AB - BACKGROUND AND PURPOSE: To improve the assessment of cerebral vasoreactivity using acetazolamide (ACZ), we studied the time course of the response and the relationship between dose, response, and serum concentration. METHODS: Blood flow velocities were measured with the use of transcranial Doppler ultrasonography in one of the middle cerebral arteries of 48 healthy subjects after the intravenous administration of 1 to 1.6 g ACZ. In 34 subjects (group 1), velocities were measured every second minute to detect the maximum middle cerebral artery velocity increase. We also measured regional cerebral blood flow using single photon emission computed tomography in 27 of the subjects in group 1 before and approximately 15 to 20 minutes after the ACZ injection. The serum concentration of ACZ was measured in 15 subjects. In the remaining 14 subjects (group 2), middle cerebral artery velocity measurements were made 10, 25, 30, and 45 minutes after ACZ administration to obtain information regarding the late time course of the response. RESULTS: In group 1 the plateau phase of the velocity response was reached 8 to 15 minutes after ACZ administration. A large range of velocity increase was observed, and a significant correlation was found between the maximum velocity increase and the dose and serum concentration of ACZ. In group 2 subjects, maximum velocities were maintained 30 minutes after the injection, but after 45 minutes velocities had decreased to 68% of their highest level. No significant relationship was found between dose or serum concentration of ACZ and the regional cerebral blood flow increase. The velocity increase after ACZ was similar in both older and younger subjects. CONCLUSIONS: This study shows that cerebral vasoreactivity is best assessed 10 to 30 minutes after ACZ administration and that the dose should probably exceed 15 mg/kg if a maximum vasodilatory response in the cerebral circulation is to be obtained. PMID- 7491656 TI - Hyperbaric oxygen after global cerebral ischemia in rabbits reduces brain vascular permeability and blood flow. AB - BACKGROUND AND PURPOSE: Hyperbaric oxygen (HBO) has been advocated as a therapy to improve neurological recovery after ischemia, since HBO may improve tissue oxygen delivery. We examined the effect of HBO treatment after global cerebral ischemia on early brain injury. METHODS: Rabbits were subjected to 10 minutes of global cerebral ischemia by cerebrospinal fluid compression. After 30 minutes of reperfusion, rabbits either were subjected to HBO for 125 minutes and then breathed 100% O2 at ambient pressure for 90 minutes or breathed 100% O2 for 215 minutes. At the end of reperfusion and 90 minutes after exposure, brain vascular permeability and cerebral blood flow were measured. Somatosensory evoked potentials were monitored throughout the experiment. RESULTS: HBO treatment reduced (P < .05) brain vascular permeability by 16% in gray matter and by 20% in white matter. Cerebral blood flow was lower (P < .05) in the HBO group (40.9 +/- 1.9 mL/min per 100 g, mean +/- SEM) compared with controls (50.8 +/- 2.0 mL/min per 100 g). Somatosensory evoked potential recovery was similar in the two groups (P > .05). CONCLUSIONS: HBO administered after global cerebral ischemia promoted blood-brain barrier integrity. HBO treatment also reduced cerebral blood flow; this effect was not associated with a reduction in evoked potential recovery. Since neurological outcome after global cerebral ischemia is generally poor and treatment options are limited, HBO should be further investigated as a potential therapy. PMID- 7491658 TI - Activity of smooth muscle phosphatases 1 and 2A in rabbit basilar artery in vasospasm. AB - BACKGROUND AND PURPOSE: Subarachnoid hemorrhage frequently leads to a long-term cerebral artery narrowing called vasospasm. Recently, the involvement of myosin light chain kinase has been found in experimental vasospasm in our laboratory. We therefore measured the activity of serine/threonine protein phosphatases 1 and 2A in the rabbit basilar artery in vasospasm and in vasocontraction to study their role, particularly in regard to vasospasm compared with vasocontraction. METHODS: Vasospasm was produced in the rabbit basilar artery by a two-hemorrhage method. Vasocontraction was induced by local application of KCl or serotonin to the rabbit basilar artery after a transclival exposure. The control animals were treated with saline instead of fresh blood. Serine/threonine protein phosphatase activity in the basilar artery was assayed with the use of [32P]phosphorylase-a as a substrate; protein phosphatase 1 activity was evaluated as protein phosphatase activity in the presence of 1 nmol/L okadaic acid, whereas protein phosphatase 2A activity was assessed as protein phosphatase activity inhibited by 1 nmol/L okadaic acid. RESULTS: Values of mean activity of protein phosphatase 1 in myofibrillar extract were 3.58 +/- 0.26 nmol/min per milligram in the control group, 3.22 +/- 0.12 nmol/min per milligram in the spastic group on day 2, and 3.01 +/- 0.16 nmol/min per milligram in the spastic group on day 4 (a significant decrease in protein phosphatase 1 activity in the spastic group on days 2 and 4). In contrast, these values did not show any significant changes in the KCl and serotonin groups. Values of mean activity of protein phosphatase 2A in cytosolic extract were 0.90 +/- 0.07 nmol/min per milligram in the control group, 0.75 +/- 0.10 nmol/min per milligram in the spastic group on day 2, and 0.62 +/- 0.17 nmol/min per milligram in the spastic group on day 4 (a significant reduction in protein phosphatase 2A in the spastic group on days 2 and 4). There was no evidence of significant changes of protein phosphatase 2A in cytosolic extract in the KCl and serotonin groups. CONCLUSIONS: Protein phosphatase 1 in myofibrillar extract is reported to catalyze the dephosphorylation of myosin light chain and calponin, whereas protein phosphatase 2A in cytosolic extract catalyzes the dephosphorylation of calponin and caldesmon. In addition, the phosphorylation of calponin and caldesmon results in the loss of their ability to inhibit smooth muscle contraction. Therefore, the significant decrease in activity of protein phosphatases 1 and 2A in vasospasm may result in uninterrupted vascular smooth muscle contraction by the preservation of phosphorylation of not only myosin light chain but also calponin and caldesmon. PMID- 7491657 TI - HU-211, a novel noncompetitive N-methyl-D-aspartate antagonist, improves neurological deficit and reduces infarct volume after reversible focal cerebral ischemia in the rat. AB - BACKGROUND AND PURPOSE: HU-211 is a nonpsychotropic cannabinoid analogue that has been shown to act as a functional N-methyl-D-aspartate receptor blocker. We investigated the neuroprotective efficacy of HU-211 in a model of reversible middle cerebral artery occlusion (MCAo) in rats. METHODS: Male Wistar rats were anesthetized with halothane and subjected to 90 minutes of temporary MCAo by retrograde insertion of an intraluminal nylon suture, coated with poly-L-lysine, through the external carotid artery into the internal carotid artery and MCA. The drug (HU-211 in cosolvent, 4 mg/kg IV) or vehicle was administered in a blinded fashion 70 minutes after onset of MCAo. Behavioral tests were evaluated during occlusion (60 minutes) and for a 3-day period after MCAo. Three days after MCAo, brains were perfusion-fixed, and infarct volumes were determined. RESULTS: HU-211 significantly improved the neurological score compared with vehicle during the 3 days after MCAo. Treatment with HU-211 also significantly reduced both infarct volume (mean +/- SEM, 66.6 +/- 12.5 versus 149.8 +/- 36.3 mm3) and brain swelling (2.61 +/- 1.33% versus 6.66 +/- 1.24%) compared with vehicle-treated rats (n = 17 in each group). CONCLUSIONS: These results demonstrate the neuroprotective ability of HU-211 in focal cerebral ischemia as judged by neurological score, infarct size, and brain swelling. Reversible MCAo with the use of a poly-L-lysine coated intraluminal suture proved to be a reliable and effective modification of this technique, yielding consistent results. PMID- 7491659 TI - Immunohistochemical detection of Ki-67 in replicative smooth muscle cells of rabbit carotid arteries after balloon denudation. AB - BACKGROUND AND PURPOSE: Ki-67 immunohistochemistry has been shown to be useful for paraffin sections from human species after wet heating. We applied Ki-67 immunohistochemistry to rabbit arteries after balloon denudation and compared the results with proliferating cell nuclear antigen (PCNA) immunohistochemistry. METHODS: Monoclonal antibodies MIB-1 for Ki-67 and PC-10 for PCNA were used to detect replicative smooth muscle cells in rabbit carotid arteries during a period of 4 weeks after balloon endothelial denudation. RESULTS: We demonstrated clear immunoreactivities for Ki-67 in paraffin sections of rabbit arteries after hydrated autoclaving. Ki-67-positive smooth muscle cells appeared throughout the thickened intima 1 week after endothelial denudation. At 2 weeks, Ki-67-positive cells were confined to the surface layer of the intima. PCNA-positive cells appeared in almost the same location by 2 weeks after endothelial denudation but were significantly greater in number than Ki-67-positive cells. PCNA-positive cells remained in the surface layer of the intima 4 weeks after endothelial denudation, while Ki-67-positive cells had almost disappeared from the intima. CONCLUSIONS: Our results indicate that Ki-67 immunohistochemistry using MIB-1 monoclonal antibody provides a powerful tool, even in rabbit species, for the detection of replicative smooth muscle cells during the repair of arterial injury and that it detects replicative cells more accurately than PCNA immunohistochemistry. PMID- 7491661 TI - Nonocclusive common carotid artery thrombosis in the rat results in reversible sensorimotor and cognitive behavioral deficits. AB - BACKGROUND AND PURPOSE: Microemboli released during transient ischemic attack, stroke, and cardiac surgery are thought to cause a variety of functional deficits in humans. The purpose of this study was to characterize the type and extent of neurobehavioral deficits present after photochemically induced common carotid artery thrombosis (CCAT), a thromboembolic model of stroke in the rat that results in a platelet emboli shower. METHODS: Thirty-two male Wistar rats were assigned to four groups. Groups 1 (n = 8) and 3 (n = 8) were long-term (6-week survival) and short-term (2-week survival) experimental groups subjected to right CCAT with the use of the photochemical technique. Groups 2 (n = 8) and 4 (n = 8) served as sham-operated controls for each experimental group. A battery of behavioral tests was applied daily beginning 24 hours after thrombosis; this consisted of elicited forelimb placing, postural reflex, beam balance, beam walking, and open field activity. Cognitive testing with a water maze task was performed on post-CCAT days 30 to 33 for groups 1 and 2 and on post-CCAT day 2 for groups 3 and 4. Ten-micrometer coronal brain sections were stained with hematoxylin and eosin, and infarct location and frequency were determined. RESULTS: Significant sensorimotor deficits were observed, which recovered within 2 weeks after CCAT. The data that follow are derived by combining the two experimental groups and comparing these with the two sham groups. The following tests showed significant effects after CCAT: contralateral elicited forelimb placing, ipsilateral elicited forelimb placing, beam balance, and beam walking score. Cognitive dysfunction was seen acutely (group 3 animals) at 2 days after CCAT; Morris water maze length and latency to target were significantly greater in the experimental group. No deficits were seen in postural reflex, open field activity, or delayed cognitive testing. Histopathological assessment revealed small infarcts in 11 of 16 thrombosed rats. However, a strong relationship between neurobehavioral deficits and infarct location was not consistently demonstrated. CONCLUSIONS: CCAT produces consistent sensorimotor and cognitive behavioral deficits that recover within 2 weeks of injury. Behavioral outcome was not necessarily associated with overt histopathological damage, suggesting that reversible injury mechanisms, both vascular and neuronal, may be partly responsible for the temporary loss of function. These data strengthen the role of CCAT as a clinically relevant model of thromboembolic stroke. PMID- 7491660 TI - Differential changes in alpha- and beta-adrenoceptors in the cerebral cortex and hippocampus of the Mongolian gerbil after unilateral brain ischemia. AB - BACKGROUND AND PURPOSE: Changes in adrenoceptors in the cerebral cortex and hippocampus of Mongolian gerbils after brain ischemia were investigated. METHODS: Twenty-four hours after unilateral occlusion of the common carotid artery, alpha 1-, alpha 2-, and beta-receptors of the membrane fraction of the cerebral cortex or the hippocampus were analyzed by binding assay with the use of [3H]prazosin, [3H]p-aminoclonidine, and [125I]cyanopindolol as radioligands, respectively. RESULTS: In the cerebral cortex, the number of binding sites (Bmax) and the dissociation constant (Kd) of [3H]prazosin were not altered, whereas the Bmax value of [3H]p-aminoclonidine binding was decreased by 30% and that of [125I]cyanopindolol binding by 16% without a change in Kd values for the ligands. In the hippocampus, the Bmax values of [3H]prazosin, [3H]p-aminoclonidine, and [125I]cyanopindolol bindings were decreased by 21%, 53%, and 19%, respectively, but there was no change in the Kd values for the ligands. The bindings of [3H]prazosin and [3H]p-aminoclonidine of the contralateral side of the cerebral cortex and the hippocampus were not altered by ischemia, but that of [125I]cyanopindolol was decreased when compared with normal tissues. CONCLUSIONS: These results show that ischemia results in a decrease in brain alpha 1-, alpha 2 , and beta-adrenoceptors to various degrees, depending on the brain area and the types of receptors, and suggest that vulnerability of the brain to ischemia is different depending on brain areas and that the regulatory mechanisms of alpha 1 , alpha 2-, and beta-receptors are different. PMID- 7491662 TI - Characterization of an anterior circulation rat subarachnoid hemorrhage model. AB - BACKGROUND AND PURPOSE: Our aim was to demonstrate the feasibility of an angiographically controlled rat model for the study of macrocirculatory and microcirculatory changes of the anterior intracranial circulation after subarachnoid hemorrhage. METHODS: Subarachnoid hemorrhage was induced by transorbital injection of 0.3 mL of nonheparinized autologous arterial blood into the chiasmatic cistern. Changes in regional cerebral blood flow were continuously recorded with the use of laser-Doppler flowmetry over the parietal cortex. Angiographic verification of middle cerebral artery diameter was performed by carotid catheterization at baseline and 2 days after injection of blood or artificial cerebrospinal fluid. We monitored intracranial and systemic blood pressure during and after injections. RESULTS: Injection of artificial cerebrospinal fluid in the control group did not change the diameter of the middle cerebral artery. Injection of blood caused a significant arterial narrowing of 17.5%, from 0.37 +/- 0.04 mm to 0.31 +/- 0.04 mm after 2 days (P = .0001). In the control group regional cerebral blood flow decreased to 75.9 +/- 16.8% of preinjection control but quickly recovered to 99.7 +/- 19.4%. Intracranial pressure increased for 5 minutes after the injection to a maximum of 27.3 +/- 8.9 mm Hg, accompanied by a 10% decrease in mean arterial pressure. A fall in cerebral blood flow to 53.1 +/- 26.3% in blood-injected animals that recovered to only 80.7 +/- 16.9% of baseline values during the observation period of 30 minutes was noted. A peak intracranial pressure of 45.7 +/- 11.5 mm Hg occurred 2 minutes after injection with a decrease in mean arterial pressure of 13%, resulting in a markedly lower cerebral perfusion pressure than in the control group. CONCLUSIONS: An angiographically controlled model of subarachnoid hemorrhage primarily involving the anterior circulation is feasible in the rat. The resulting narrowing of the middle cerebral artery reflects moderate vasospasm and will allow further microcirculatory studies with cranial windows. PMID- 7491663 TI - Perfusion imaging of the brain by B-mode ultrasonography. An experimental study in rabbits. AB - BACKGROUND AND PURPOSE: The purpose of this study was to assess the ultrasonic imaging of normal rabbit brain by using echocontrast enhancer to identify the brain tissue perfusion. METHODS: A solution of SH/TA-508 (containing microbubbles of mean size 2 to 4 microns; n = 4) was used for contrast enhancement of rabbit brain tissue and was administered through the left internal carotid artery. Contrast enhancement in the ipsilateral parietal area was examined by placing a 7.5-MHz transducer on the dura after a craniectomy. Throughout the experiment, we measured parameters including arterial blood pressure, arterial blood gases, and serum glucose and hematocrit levels. We investigated whether the administration of an echocontrast enhancer would be safe for the brain tissue (n = 4). RESULTS: After injection of 100 mg/mL SH/TA-508, contrast enhancement was clearly observed within approximately 1 second in the left hemisphere (especially in the left cerebral cortex) in all animals and disappeared within 3 seconds after the end of injection. All physiological parameters remained within the normal range throughout the experiment. No focal abnormalities were observed on hematoxylin eosin staining of the brain tissue. CONCLUSIONS: Contrast enhancement was observed in the ipsilateral hemisphere with the administration of SH/TA-508. Contrast enhancement on B-mode brain imaging would be useful for real-time identification of brain tissue perfusion and should become a promising method for bedside clinical assessment of brain tissue perfusion. PMID- 7491664 TI - Effect of acetazolamide on regional cerebral oxygen saturation and regional cerebral blood flow. AB - BACKGROUND AND PURPOSE: To verify whether the monitoring of regional cerebral oxygen saturation (rSO2) with transcranial near-infrared spectroscopy would successfully reflect changes in intracranial hemodynamics but not changes in extracranial compartment, we measured rSO2 and regional cerebral blood flow (rCBF) simultaneously in seven patients with cerebral ischemia and five normal volunteers before and after acetazolamide administration. SUMMARY OF REPORT: The baseline values of rSO2 and rCBF were 64.2 +/- 5.6% and 53.9 +/- 11.1 mL/100 g per minute, respectively. rCBF increased by 44.4 +/- 23.3% and rSO2 significantly increased to 69.6 +/- 5.6% after acetazolamide administration. Bilateral simultaneous measurement of rSO2 indicated a tendency that the larger the delta rSO2, the greater the delta%rCBF. The relationship between rSO2 level and rCBF value fit significantly on the theoretical curve calculated from Fick's equation. CONCLUSIONS: It is suggested that monitoring of rSO2 with INVOS-3100 could be a useful indicator in the evaluation of intracranial hemodynamic changes. PMID- 7491665 TI - Transient ischemic attacks and minor stroke during sleep. Relationship to obstructive sleep apnea syndrome. AB - BACKGROUND: It is reported that 13% to 44% of all cerebrovascular accidents (CVAs) occur during sleep. In addition to other well-known risk factors, snoring, sleep apnea, obesity, and daytime sleepiness have been shown to significantly increase the risk of stroke. We describe two cases that support the statistical relationship between snoring, sleep apnea, and CVA during sleep. CASE DESCRIPTIONS: In the first case, motor aphasia was noted in a 64-year-old, 5-ft, 1-in, 218-lb woman when she awakened from sleep at approximately 4 AM. This completely resolved within 3 hours. During her subsequent hospitalization she was found to have severe obstructive sleep apnea that responded well to treatment with nasal continuous positive airway pressure. There has been no recurrence of symptoms in this patient. The second patient was a 59-year-old, 5-ft, 6-in, 260 lb woman who presented to the Sleep Disorders Center with signs and symptoms of severe sleep apnea. In addition, she had awakened from sleep approximately 6 months earlier with numbness and weakness on her right side. Although these symptoms had greatly improved, she continued to complain about residual weakness that was worse on awakening from sleep. Sleep studies confirmed severe obstructive sleep apnea that responded very well to treatment with nasal continuous positive airway pressure. CONCLUSIONS: Snoring and obstructive sleep apnea not only increase the statistical risk of CVA but could be the proximal trigger that precipitates these events during sleep. These two cases provide clinical support for this relationship. Successful diagnosis and treatment of obstructive sleep apnea in the patient with transient ischemic attacks and minor stroke may be an important tool for preventing recurrence. PMID- 7491666 TI - Leptomeningeal dissemination of malignant glioma simulating cerebral vasculitis. Case report with angiographic and pathological studies. AB - BACKGROUND: The complex clinical and radiological picture of leptomeningeal spread of tumor is well recognized as a problem of systemic cancer but is less frequent in primary cerebral glioma, particularly as a presenting picture. While brain ischemia and infarction may occur in patients with subarachnoid tumor, the mechanism for these complications remains unclear. Angiographic and pathological demonstrations of direct vascular involvement by disseminated glioma are particularly sparse. We report a patient presenting with multiple infarctlike lesions with postmortem evidence of direct vascular involvement by glioma. CASE DESCRIPTION: A 54-year-old woman presenting with seizures, headache, and changes in mental status was found to have vascular narrowing in cerebral blood vessels and ischemic lesions on neuroimaging studies of the brain, interpreted as cerebral vasculitis. A brain biopsy showed leptomeningeal glioma. Postmortem examination demonstrated a glioblastoma arising around the right sylvian fissure with extensive subarachnoid dissemination of tumor. The leptomeningeal tumor caused vascular narrowing by encasement, direct vascular wall invasion, and thrombosis and was associated with underlying infarctlike foci of parenchymal necrosis. CONCLUSIONS: This case demonstrates an unusual presentation of glioblastoma clinically and radiographically mimicking cerebral vasculitis, and it illustrates a variety of mechanisms for tumor-produced vascular compromise. PMID- 7491667 TI - Steal and cerebral arteriovenous malformations. PMID- 7491668 TI - Comparison of static and dynamic cerebral autoregulation measurements. PMID- 7491669 TI - Carotid diameter, wall thickness, and parietal stress in essential hypertension. PMID- 7491670 TI - Mitral annular calcification versus mitral valve calcification. PMID- 7491671 TI - Plasma homocysteine in ischemic stroke. PMID- 7491672 TI - Apolipoprotein E variants in ischemic stroke. PMID- 7491673 TI - Liposomal cyclosporine. Comparison of drug and lipid carrier pharmacokinetics and biodistribution. AB - In a preceding paper (Ouyang et al., 1995, this issue), we have characterized cyclosporine incorporation into well-defined liposomal systems, large unilamellar vesicles. This study demonstrated that only modest drug levels could be accommodated within the membrane, particularly for cholesterol-containing liposomes, and that rapid drug exchange could occur between vesicles. This raised the possibility that following intravenous administration, drug migration to other blood components might negate the potential benefits arising from liposomal delivery. We have, therefore, examined the pharmacokinetics and biodistribution of both cyclosporine and its liposomal carrier. We show that whereas liposomes, as expected, are only slowly cleared from the blood, redistribution of cyclosporine occurs much more rapidly. Further we have shown that liposomal loss of cyclosporine in blood results from drug migration to the lipoproteins and, to a lesser extent, the erythrocytes. As a result, while liposomes accumulate preferentially in organs of the reticuloendothelial system after intravenous administration, tissue cyclosporine levels, in general, do not reflect the distribution profile obtained for the liposomal carrier. PMID- 7491674 TI - Increased granzyme B mRNA after alloincompatible myoblast transplantation. AB - Normal C57BL/10SnJ myoblasts were transplanted into the tibialis anterior of C57BL/10SnJ, C57BL/ScSn mdx, or BALB/c mice. These transplantations allowed us to investigate the immune response not only against MHC but also against dystrophin introduced in the dystrophic muscles by such transplantations. Recently, our group reported following myoblast transplantations cellular infiltration of the host muscle by class II MHC cells, macrophages, and lymphocytes expressing CD4 or CD8 and IL-2 receptors. In the present study, activation of these infiltrating lymphocytes was investigated by measuring the expression of granzyme B mRNA. We used reverse-transcriptase polymerase chain reaction to detect granzyme B mRNA at various intervals after myoblast transplantations. To standardize the results, the mRNA were reverse transcribed using an oligo (dt) so that beta-actin mRNA could also be amplified from the same cDNA preparation. Granzyme B mRNA was increased for at least 3 weeks after MHC alloincompatible grafts. The absence of increased granzyme B expression after allocompatible transplantation in mdx mice suggests that dystrophin is not sufficiently immunogenic to induce short term acute rejection. These results indicate that lymphocytes infiltrated in muscles injected with histoincompatible myoblasts are activated and sustain the requirement for an adequate immunosuppression after such transplantations. PMID- 7491675 TI - IgG donor-specific crossmatches are not associated with graft rejection or poor graft survival after liver transplantation. An assessment by cytotoxicity and flow cytometry. AB - Early studies in liver transplantation suggested that there was no association between graft outcome or rejection and the presence of alloantibodies before transplantation. More recent reports have suggested lower graft survival rates and a higher incidence of chronic rejection in patients with IgG warm-T crossmatches. In the present study, panel reactive antibody, direct crossmatch testing, and flow cytometry were used to detect preformed antibodies in sera from 158 consecutive adult recipients of first hepatic grafts. The relationship between preformed antidonor antibodies and liver allograft survival and rejection was determined. Twenty-six (17%) patients were panel reactive antibody (PRA) positive before transplantation, 22 (15%) had positive donor-specific crossmatches, and 14 (9%) were positive by IgG-specific flow cytometry. Cumulative survival distribution and multivariate analysis failed to reveal any significant associations between overall graft survival and antibody status. Graft survival in patients with PRA-positive sera was 81% compared with 77% for those with PRA-negative sera, 68% for those with positive donor-specific crossmatches compared with 80% for those who were donor-specific crossmatch negative, and 79% for those who were antibody positive by flow cytometric analysis compared with 78% for those who were antibody negative. Subgroup analysis also failed to reveal any significant associations. In addition, Cox proportional hazards regression analysis failed to reveal a relationship between acute or chronic graft rejection with the presence or absence of preformed antibodies, irrespective of immunoglobulin class, cell type (T or non-T), specificity, or technique used for antibody detection. In conclusion, there appears to be no association between either donor-specific or "third-party" alloreactive IgG or IgM antibodies and liver transplant survival or rejection. These data do not indicate a need for prospective crossmatching of liver transplant recipients. PMID- 7491676 TI - Direct recognition of SLA- and HLA-like class II antigens on porcine endothelium by human T cells results in T cell activation and release of interleukin-2. AB - To investigate whether human T cells can directly recognize pig xenoantigens, highly purified human CD4+ and CD8+ T cells were incubated with pig aortic endothelial cells (PAEC). The response was measured by [3H]thymidine uptake and release of bioactive interleukin-2. A detailed examination of MHC expression by cultured PAEC and tissue sections of porcine aorta and heart showed porcine endothelial cells (EC) to be constitutively positive for SLA class II and antigens that crossreact with HLA class II molecules. Low level expression of B7 receptors was detected by binding of both human and mouse CTLA-4-Ig to untreated PAEC, which was enhanced significantly by treatment with recombinant porcine interferon-gamma. Human T cells, purified by positive selection and residual DR+ cells removed by lymphocytolysis, were shown to be functionally free of monocytes. Untreated PAEC elicited strong proliferation by human CD4+ T cells: CD8+ T cells also proliferated, but more weakly. This response was inhibited by CTLA-4-Ig. Blocking studies were performed with mAbs that bind to PAEC and not human EC (MSA3, TH16B), an mAb that binds to human and porcine EC (DA6.231), and L243, which binds to human and not porcine EC. The proliferative response of CD4+ T cells to PAEC was inhibited significantly by mAbs against swine and human determinants. In contrast, the response of CD4+ T cells to human EC was inhibited only by mAbs against human determinants. Experiments that directly compared the CD4+ and CD8+ T cell responses to PAEC and the human EC line EAhy.926, both with and without prior treatment with species-specific interferon gamma, demonstrated greater proliferation and 5-10 times more interleukin-2 in response to pig EC than to human EC. PMID- 7491677 TI - HLA-DR matching in corneal transplantation. Systematic review of published evidence. Corneal Transplant Follow-up Study Collaborators. PMID- 7491678 TI - Methodology for the assessment of lung protection. Human pulmonary artery endothelial cell preservation using haemaccel. AB - This investigation was designed to show an original methodology for the assessment of lung preservation and to analyze the efficacy of a low potassium polygelin solution (haemaccel [HM]) on isolated human pulmonary artery endothelial cells. The effects of HM were compared with those of low potassium dextran (LPD), Belzer (University of Wisconsin [UWS]), and Euro-Collins solutions. The viability of the endothelial cultures was assessed by means of both total protein content and recovery of metabolic cellular function expressed as the protein synthesis rate after 6 hr and 16 hr of incubation at 10 degrees C. Our results failed to show any significant difference in the total protein content for HM, LPD, and UWS, both after 6 and 16 hr of incubation; however, the Euro-Collins-preserved sample revealed a significant drop in this parameter as early as 6 hr after the start. This finding was regarded as a clear indication of cellular cytotoxicity. In contrast, the metabolism recovery capacity of the cells varied significantly between HM and UWS at 6 hr and among HM, LPD, and UWS at 16 hr; at 6 hr, however, no significant difference was observed between HM and LPD. In conclusion, HM appears to exert a more significant effect on human pulmonary artery endothelial cell metabolism recovery than do the other fluids, thus suggesting its suitability as a long-term pulmonary perfusate. PMID- 7491680 TI - Laparoscopic live donor nephrectomy. AB - A laparoscopic live-donor nephrectomy was performed on a 40-year-old man. The kidney was removed intact via a 9-cm infraumbilical midline incision. Warm ischemia was limited to less than 5 min. Immediately upon revascularization, the allograft produced urine. By the second postoperative day, the recipient's serum creatinine had decreased to 0.7 mg/dl. The donor's postoperative course was uneventful. He experienced minimal discomfort and was discharged home on the first postoperative day. We conclude that laparoscopic donor nephrectomy is feasible. It can be performed without apparent deleterious effects to either the donor or the recipient. The limited discomfort and rapid convalescence enjoyed by our patient indicate that this technique may prove to be advantageous. PMID- 7491679 TI - Cryopreservation of purified bovine islets of Langerhans. AB - In this study, bovine islets were isolated by collagenase digestion and density gradient purification, equilibrated stepwise with 3 M dimethylsulfoxide at 24 degrees C, nucleated at -150 degrees C, slow cooled at 0.25 degrees C/min down to -40 degrees C, and finally stored at -150 degrees C. After variable periods of time, the islets were quickly thawed at 37 degrees C, and dimethylsulfoxide was removed by 0.75 M sucrose. Postthawing recovery was 86 +/- 6% islet equivalents. Histology confirmed the identity and morphological integrity of the islets. Insulin release from the frozen-thawed islets was 0.13 +/- 0.03 microU/is/min at 3.3 mmol/L glucose and increased significantly (0.27 +/- 0.04 microU/is/min, P < 0.05) at 25 mmol/L glucose. Encapsulated, cryopreserved islets reversed hyperglycemia in diabetic mice after 6-8 days following implantation. Therefore, the method described in this paper permitted successful cryopreservation of bovine islets of proven in vitro and in vivo viability. PMID- 7491681 TI - Hepatitis C-associated cryoglobulinemia after liver transplantation. AB - Mixed cryoglobulinemia is well known to be associated with hepatitis C virus (HCV) infection. We report two cases in which cryoglobulinemia appeared or became grossly exacerbated after orthotopic liver transplantation. In both cases, there was co-appearance of cryoglobulinemia with the reinfection of the grafted liver with HCV. It is postulated that the cryoglobulinemia might be related to secondary HCV infection in these patients. PMID- 7491682 TI - Fulminant hepatic failure following varicella-zoster infection in a child. A case report of successful treatment with liver transplantation and perioperative acyclovir. PMID- 7491683 TI - Importance of surveillance mycobacterial cultures after liver transplantation. AB - The routine use of isoniazid prophylaxis after liver transplantation is a controversial issue because the benefits must be weighed against the risk of hepatotoxicity. We decided not to institute isoniazid prophylaxis but to study the efficacy of a surveillance mycobacterial program. One hundred patients were included in the protocol. Sputum and urine samples were processed before transplantation and on days 15, 30, 60, 90, 12, 150, and 180 for acid-fast stain and culture. One case of tuberculosis was promptly identified and successfully treated. Cases of tuberculosis with negative surveillance cultures were not identified. Our approach indicates that surveillance mycobacterial cultures can permit rapid identification of tuberculosis after liver transplantation and it is an alternative for groups who questioned isoniazid prophylaxis. PMID- 7491684 TI - Varicella vaccine in children requiring renal or hepatic transplantation. PMID- 7491685 TI - Metabolic complications after liver transplantation. Diabetes, hypercholesterolemia, hypertension, and obesity. AB - We retrospectively studied the incidence of diabetes, hypercholesterolemia, hypertension, and obesity in 123 consecutive adult liver transplant recipients (61 men and 62 women) who were alive at least 1 year after transplantation. We also studied the change in these metabolic complications in 61 patients who subsequently were able to be tapered to 5 mg prednisone per day. One year after transplantation--a point at which almost all patients were on maintenance immunosuppression and had stable graft function--the incidence of diabetes was 13% and hypertension was 69.1%. The overall incidence of hypercholesterolemia (serum cholesterol > 240 ng/ml) was 31% and was more frequent in women than in men (38.7% vs. 23.0%, P < 0.06). The incidence of obesity at 1 year was 41.9% in women and 39.3% in men. With tapering of prednisone from 10 mg to 5 mg per day in 61 patients, the mean serum cholesterol decreased from 224.6 +/- 65.2 mg/dl to 203.3 +/- 65.5 mg/dl, P < 0.005. With steroid tapering, 8 patients were able to discontinue antihypertensive medications and 4 were able to discontinue insulin treatment for diabetes. Five patients became obese during the steroid-tapering period. No patient developed irreversible rejection with steroid tapering and no immunologic graft losses occurred more than a year after transplantation. Nine patients who lived a year subsequently died. Of these, 7 patients were diabetic and 2 died of cardiac disease. We conclude that metabolic complications such as diabetes, hypertension, and hypercholesterolemia are common later after liver transplantation and that these may contribute to patient morbidity and mortality. In addition, we conclude steroid tapering to 5 mg/day does not lead to graft loss and may decrease the incidence and severity of late metabolic complications. PMID- 7491686 TI - Intrathymic injection of alloantigen may lead to hyperacute rejection and prolonged graft survival of heart allografts in the rat. PMID- 7491687 TI - Requesting organ donation from brain-dead victims versus requesting tissue donation from patients after cardiac arrest. PMID- 7491689 TI - Long-term effects of small bowel transplantation on intestinal motility. AB - We previously found that the contractile motility of the jejunum was increased 4 weeks after transplantation, and that the dominant intrinsic neural component was changed from cholinergic to nonadrenergic, noncholinergic (NANC). The present study investigated the long-term effects of transplantation on jejunal motility using rats that survived for 2 years after surgery. Jejunal strips were harvested from various groups of rats, and intestinal motility was assessed by electrical transmural stimulation. Stimulation produced a similar increase of contraction at 4 weeks and 2 years after grafting. Pretreatment with atropine showed that the cholinergic component of contraction was 45%, 24%, 32%, and 24% in controls, rats 4 weeks after transplantation, 2-year-old controls, and rats 2 years after transplantation, respectively. The NANC component (obtained with atropine and guanethidine) in each group was, respectively, 56%, 73%, 60%, and 69%. The actual value of the tetrodotoxin-insensitive myogenic component was significantly increased at 2 years after transplantation. A substance P antagonist ([Arg6, D Trp7,9, Mephe8], substance P 6-11), inhibited most of the NANC contraction after transplantation. These results suggested that substance P has a key role in the motility of transplanted small bowel throughout the life of the grafts. PMID- 7491688 TI - In vivo evaluation of timing, degree, and distribution of bacterial translocation following experimental small bowel transplantation. AB - These studies were designed to evaluate the correlation between morphologic and functional changes after heterotopic auxiliary small bowel isograft with systemic venous drainage and two ostomies in 20 Lewis rats. Morphologic damage of the graft was scored by full-thickness biopsies before surgery and 1, 3, 5, and 7 days after transplant. Functional evaluation of the graft was done, at the same time points, by urinary excretion of lactulose and mannitol injected in the proximal ostomy. The intestinal permeability was also studied by injecting Escherichia coli labeled with indium-111 oxine in the proximal ostomy. Translocation of radiolabeled bacteria was quantitated in extraintestinal tissues by radionuclide counts and number of viable organisms and in vivo by scintigraphic imaging. One day after transplant, significant graft damage (score 17.2 +/- 4.2) was observed when compared with the pretransplant value (7.3 +/- 2.6). The degree of tissue injury was similar on days 3 (15.8 +/- 3.5) and 5 (16.1 +/- 3.9) after transplant and remained high on day 7 (11.8 +/- 2.8). The lactulose to mannitol ratio showed a significantly increased permeability on day 1 (17.5) versus pretransplant values (2.6), remained high on day 3 (8.6), and returned to normal values on day 5 (2.8). Translocation of bacteria to distant organs, as measured by both radionuclide counts and number of viable organisms, was strikingly enhanced on day 1 after transplantation, compared with control animals, but returned to the pretransplant value on day 3. A good qualitative and quantitative correlation was observed between radionuclide counts in the extraintestinal organs and in vivo images obtained by scintigraphic scanning. In conclusion, in this model, timing and degree of bacterial translocation do not seem to correlate well, with the exception of the acute post-transplantation phase, with morphologic and permeability changes of the graft. Evaluation of translocation by scintigraphic imaging appears a suitable approach to study in vivo the kinetics and distribution of this process. PMID- 7491690 TI - Investigation of the potential use of immunosuppressive agent gliotoxin in organ transplantation. AB - Gliotoxin is an immunosuppressive secondary metabolite produced by several pathogenic fungi. It has previously been shown to prevent graft-versus-host disease in transplantation of allogeneic mouse bone marrow and to reduce the immunogenicity of human fetal pancreas. We here report on the effect of gliotoxin on the prevention of rejection of allografts in two distinct models. Bathing mouse thyroid tissue in gliotoxin solution for 16 hr prolonged graft survival following transplantation into allogeneic recipients. In contrast the perfusion of rat kidneys with gliotoxin followed by 1 hr of incubation before orthotopic transplantation had little success with preventing allograft rejection. This disparity is most likely due to the incubation in the renal model not allowing sufficient time for the elimination of antigen presenting cells in the donor organ. However, the success with the thyroid grafts demonstrates the potential of gliotoxin as an immunomodulating agent in organ transplantation and warrants further investigation in other systems. PMID- 7491691 TI - Use of intravenous immunoglobulin to delay xenogeneic hyperacute rejection. An in vivo and in vitro evaluation. AB - The delaying action of intravenous immunoglobulin (IVIG) from human origin on hyperacute xenogeneic rejection was assessed in the guinea pig-to-rat combination. IVIG (1500 mg/kg) injected i.v. into Lewis rats 1 hr before grafting significantly prolonged the mean guinea pig heart survival time (167 min, P < 0.005) compared with control injections using NaCl (12 min) or the IVIG glycine vehicle (11 min). The effect of IVIG was also assessed in vitro in the pig-to human combination. A dose-dependent inhibition of the complement-mediated direct cytotoxicity of human serum on pig RBC was shown using IVIG. The weak direct cytotoxicity of IVIG to pig RBC, which was abolished after preincubating IVIG with pig RBC, was attributed to the anti-pig xenoreactive natural antibodies (XNA) contained in the IVIG preparation. In vitro, XNA-depleted IVIG exerted a significantly stronger inhibitory effect than non-XNA-depleted IVIG, suggesting the use XNA-depleted IVIG in the pig-to-human combination. Although the mechanism of the inhibitory effect of IVIG remains to be clarified, IVIG may represent a new and simple therapeutic modality against xenogeneic hyperacute rejection. PMID- 7491692 TI - Impact of renal transplantation on uremic cardiomyopathy. AB - In chronic uremia, cardiomyopathy manifests itself as systolic dysfunction, concentric left ventricular (LV) hypertrophy, or LV dilatation. To determine the impact of renal transplantation on uremic cardiomyopathy, all dialysis patients participating in a long-term cohort study who received a successful renal transplant were followed with echocardiography. The transplanted group comprised 102 of 433 (24%) endstage renal disease (ESRD) patients. They were significantly younger and, on starting ESRD therapy, had significantly less ischemic heart disease and cardiac failure than the overall ESRD cohort. During followup, ischemic heart disease developed in only 1 patient and none experienced cardiac failure. In the 12% (n = 12) of patients with systolic dysfunction before renal transplant, fractional shortening normalized in all patients, increasing from 21.5 +/- 4.6% to 33.5 +/- 5.6% after transplantation. In the 41% (n = 41) with concentric LV hypertrophy before transplantation, the LV mass index improved from 158 +/- 39 g/m2 to 132 +/- 39 g/m2. LV dilatation was present in 32% (n = 32) of patients before transplantation. After transplantation, LV volume fell from 116 +/- 3.1 ml/m2 to 89 +/- 21 ml/m2, and LV mass index in this group fell from 166 +/- 55 g/m2 to 135 +/- 37 g/m2. It was not possible to associate risk factors characteristic of the uremic state with the improvement in cardiac structure and function, although the fall in LV mass was significantly associated with fall in blood pressure. We conclude that correction of the uremic state by renal transplantation leads to normalization of LV contractility in systolic dysfunction, regression of hypertrophy in concentric LV hypertrophy, and improvement of cavity volume in LV dilatation. The degree of improvement suggests that dialysis patients with uremic cardiomyopathy would benefit from renal transplantation. PMID- 7491693 TI - Medication compliance after renal transplantation. AB - Noncompliance is known to be an important cause of late graft failure after renal transplantation. We investigated prospectively the degree of compliance with immunosuppressive and antihypertensive drugs during the first year after renal transplantation by monthly pill counts. In addition, we examined whether noncompliance was related to a number of demographic and clinical variables or to the occurrence of rejections. The study population consisted of 127 patients who were involved in a randomized trial comparing cyclosporine monotherapy with azathioprine-prednisone treatment. Average compliance rates approximated 100%, although considerable variability within and between subjects was observed. Using an arbitrary limit to classify patients as compliers or noncompliers, the following frequencies of noncompliance were observed during the study year: cyclosporine, 23%; azathioprine, 13%; prednisone, 23%; atenolol, 36%; and nifedipine, 32%. Average compliance scores for all immunosuppressive drugs were superior to those of antihypertensive medication (P < 0.001). Except for a better compliance for prednisone in men as compared with women, we found no consistent relationship between compliance on the one hand and several demographic variables, graft function, or quality of life on the other hand. Patients who developed one or more acute rejection episodes showed a higher degree of undercompliance, especially for prednisone, than patients without rejections (P < 0.01). Following the occurrence of a rejection episode, compliance scores improved significantly. Keeping in mind the limitations of the pill count method, we conclude that noncompliance with immunosuppressive drugs is not a huge problem during the first year after renal transplantation. However, it is likely that noncompliance contributes to a certain number of acute rejection episodes. PMID- 7491694 TI - Pretransplant status and patient survival following liver transplantation. AB - The current liver allocation system has been criticized, since available organs go to those who are the most critically ill. These recipients have the poorest overall survival. Identification of pretransplant risk factors for mortality would allow better allocation of donor livers. This study was a retrospective analysis of pretransplant clinical and laboratory parameters and subsequent postoperative liver transplant mortality to identify high-risk subgroups. Of 347 consecutive consecutive primary liver transplant recipients, 59 (17%) met United Network for Organ Sharing (UNOS) criteria for status 4. Pretransplant factors included liver function, coagulation, albumin and ammonia levels, renal function, the presence of ascites, and etiology of liver disease. Overall 1-year patient survival was significantly worse for the status 4 recipients (89.0% vs. 67.7%; P = 0.01). In a univariate analysis of pretransplant risk factors for all recipients, elevated creatinine (P = 0.008) and ammonia (P = 0.009), and UNOS status 4 (P = 0.01) significantly affected postoperative survival. In multivariate analysis of pretransplant risk factors for all recipients, elevated creatinine (P = 0.003) was the only factor to significantly affect postoperative survival. In UNOS status 4 patients, univariate analysis of pretransplant risk factors and their influence on patient survival demonstrated that prolonged coagulation partial thromboplastin time (P = 0.04) and a higher grade of encephalopathy (P = 0.02) significantly affected postoperative survival. Advanced encephalopathy (P = 0.009) and prolonged partial thromboplastin time (P = 0.01) were the only significant risk factors by multivariate analysis in status 4 patients. In status 4 and non-status 4 patients, we identified risk factors that adversely affected patient survival, but their predictive power was insufficient to deny transplantation. Despite the higher mortality in status 4 recipients, their long-term survival is only slightly worse than that of non-status 4 patients. Until better predictors of survival are ascertained, our data do not support limiting the use of donor livers in UNOS status 4 recipients. PMID- 7491695 TI - Hepatic granulomas following liver transplantation. Clinicopathologic features in 42 patients. AB - Liver granulomas have long been known to pose diagnostic problems for pathologists; however, their prevalence and associated etiologic factors have not been studied in liver transplant patients. We reviewed 3632 liver biopsy specimens from 563 patients at two institutions and identified 42 patients with posttransplant granulomas. A possible or probable etiologic factor was identified in 30 (71%) cases. Most were epithelioid granulomas and microgranulomas located in the parenchyma associated with hepatocyte necrosis (21 cases, 50%). Portal based granulomas were associated with recurrent primary biliary cirrhosis (5 cases, 12%), acute cellular rejection (2 cases, 4.8%), and a foreign body-type reaction (1 case, 2.4%). One case was associated with tuberculosis (2.4%), 4 cases occurred in a fatty liver (9.5%), and 8 patients had liver granulomas but no other significant abnormality. The granulomas were most frequent in the first 7 months after transplantation when the patients were biopsied more often and underwent episodes of rejection or acute hepatitis. Portal-based granulomas in this period were usually associated with acute cellular rejection. After 7 months, the frequency of granulomas as well as the number of biopsies decreased and portal-based granulomas associated with recurrent primary biliary cirrhosis were most common (5 cases, 12%). Rare, late-appearing parenchymal granulomas were also seen (3 cases) and consisted of 1 lipogranuloma and 2 cases of epithelioid granuloma. The latter were thought, in 1 patient, to be associated with parenchymal hepatocyte necrosis; the others were of unknown etiology. PMID- 7491696 TI - Liver transplantation in familial amyloidotic polyneuropathy. Follow-up of the first 20 Swedish patients. AB - Familial amyloidotic polyneuropathy (FAP) is an inherited fatal form of amyloidosis caused by mutant transthyretin. The disease is characterized by progressive peripheral and autonomic neuropathy. Most of the transthyretin is produced by the liver, and we have shown previously that the metabolic deficiency can be corrected by liver transplantation. In the present study, the clinical results from the first 20 patients who underwent liver transplantation for FAP in Sweden are evaluated. Three of the patients suffered from renal failure and underwent a simultaneous kidney transplantation. Fourteen of the 20 patients (70%) are alive 10-52 months after transplantation. The patients' nutritional status at the time of transplantation had a significant impact on mortality and morbidity (P < 0.007). Long-standing disease was another negative prognostic factor (P < 0.02). One year after transplantation, the nutritional status had improved (P < 0.02). Improvements were also noted in walking capacity and for gastrointestinal and urogenital symptoms. The results show that liver transplantation offers an effective means to treat patients with FAP. The procedure should preferably be performed before the nutritional status is poor and advanced organ dysfunction has developed. PMID- 7491698 TI - Elevated inflammatory cytokine levels in bone marrow graft rejection. AB - Graft rejection and graft failure represent major obstacles in allogeneic bone marrow transplantation (BMT). Cytokines possibly play a central role in the inflammatory and allospecific components of allograft rejection. Therefore, we evaluated inflammatory cytokine levels following BMT in 12 consecutive patients with graft rejection (GR). Seven of the patients underwent BMT from siblings (6 matched and 1 mismatched), 4 patients received bone marrow from other family members (3 mismatched and 1 matched), and 1 patient underwent HLA-matched unrelated BMT. Nine of 12 had a sex-mismatched BMT and 5/12 had an ABO-mismatched BMT. Nine of 12 underwent T cell-depleted (Campath anti-CDw52 moAb) BMT. Rejection was defined as marrow hypoplasia with a peripheral white blood cell count < 0.5 x 10(9)/L 21 days after BMT, in conjunction with the absence of donor cells by polymerase chain reaction analysis using a sex-mismatched probe and/or a tumor-specific probe (BCR/ABL). Twenty-five patients who underwent uneventful BMT with no GR served as controls. The levels of tumor necrosis factor (TNF), interleukin-6 (IL-6), and interleukin-1 (IL-1) were evaluated by a high sensitive RIA or an enzyme immunoassay. The levels of TNF and IL-6 were found to be higher in 10/12 and 7/7 evaluated GR patients, respectively, as compared with controls (P < 0.05). The level of IL-1 was high only in 2/12 patients. TNF elevation occurred in all patients immediately after GR. TNF and IL-6 levels were significantly higher for patients with early rejection (< 35 days after BMT) as compared with patients with late rejection (> 35 days after BMT) (P < 0.049 and P < 0.006, respectively). Eight patients engrafted after the second transplant (2 only transient). All 6 patients with stable engraftment are alive (4 with basic disease), while the 4 patients who did not engraft and the 2 patients with only transient engraftment died. In the 6 patients with no engraftment or only transient engraftment, the elevated TNF levels remained high; in the 6 patients who had stable engraftment after retransplant, TNF levels, but not IL-6 levels, decreased. In conclusion, a majority of the patients with GR displayed high levels of inflammatory cytokines (TNF and IL-6). Dysregulation of inflammatory cytokines may be involved in the pathogenesis of GR. PMID- 7491697 TI - Treatment of acute graft-versus-host disease with methylprednisolone and cyclosporine with or without an anti-interleukin-2 receptor monoclonal antibody. A multicenter phase III study. AB - A double-blind, placebo-controlled trial of BT563, including 13 European centers, was initiated in October 1989 to compare the efficacy of the combination of in vivo anti-CD25 mAb (BT 563), cyclosporine, and steroids versus placebo and CSA steroids in the treatment of grade II and III acute graft-versus-host disease (GVHD). Sixty-nine patients participated in the study, which excluded non genotypically identical allogeneic bone marrow transplant recipients. No statistically significant differences were observed, clinically or biologically, between the 2 groups before the onset of the treatment. Treatment responses were scored during and after the 3-week treatment period (mAb or placebo). Efficacy was evaluated on days 4, 10, 20, 30, and 60 or on any day the patient's condition was found to be deteriorating. Preceding and systemically untreated GVHD of grade I was observed in 59% of the cases. No statistically clinically significant differences between the 2 groups were observed during or upon completion of treatment in GVHD grade. Nine patients in the placebo group and 6 in the active group were withdrawn of the study. Thirteen of these 15 patients were withdrawn because of failure of GVHD therapy (9 in the placebo group and 4 in the BT563 group). At day 20 after onset of the treatment, the response rate was 63% and 70% for the placebo and BT563 groups, respectively (NS). Probability of survival at 1 year was 59% and 66% (NS) for the placebo and active groups, respectively. In conclusion, despite preliminary promising results in the treatment of steroid resistant acute GVHD, the role of first-line treatment with an in vivo anti interleukin-2 receptor mAb remains to be determined. PMID- 7491699 TI - Microangiopathy following allogeneic marrow transplantation. Association with cyclosporine and methylprednisolone for graft-versus-host disease prophylaxis. AB - A microangiopathic syndrome was observed in 3 of 14 (21%) patients receiving cyclosporine and methylprednisolone (CSA-MP) for graft-versus-host disease (GVHD) prophylaxis between January 1991 and June 1992 at our center. The syndrome consisted of neurological abnormalities, arterial hypertension, intravascular hemolysis with red cell fragmentation, and a drop in platelet counts after allogeneic bone marrow transplantation (BMT) for hematological malignancy, and it occurred around day 50 after BMT. Treatment with plasma exchanges against fresh frozen plasma resulted in a decrease of serum lactate dehydrogenase and an improvement of neurological symptoms. We compared CSA-MP patients retrospectively with patients who had received cyclosporine and methotrexate (CSA-MTX) for GVHD prophylaxis (n = 70) at our institution. All patients in both groups engrafted. Day 100 survival (80% vs. 79%) and transplant-related mortality (16% vs. 14%) were identical in the two groups. CSA-MP patients had significantly more acute GVHD II-IV (57% vs. 17%, P < 0.01). Arterial hypertension (P < 0.01) and neurological symptoms (P < 0.01) were significantly more frequent in the CSA-MP group. The 11 asymptomatic CSA-MP patients had significantly higher lactate dehydrogenase levels (P < 0.01) and lower platelet counts (P < 0.01) at 40, 60, and 100 days after BMT, which suggests the presence of a subclinical form of microangiopathy. Significantly higher plasma levels of von Willebrand factor antigen in CSA-MP patients on day 50 after BMT (P < 0.05) and absence of large von Willebrand factor multimers on gel electrophoresis in 4 of 13 (31%) CSA-MP patients compared with 0 of 14 (0%) CSA-MTX patients (P < 0.01) further suggest profound endothelial damage in patients receiving CSA-MP for GVHD prophylaxis. PMID- 7491700 TI - Mycobacterium haemophilum infections in bone marrow transplant recipients. AB - The purpose of this study was to describe the clinical presentation, treatment, and outcome of Mycobacterium haemophilum infection in patients undergoing bone marrow transplantation at a cancer center. Bone marrow transplant recipients with M haemophilum infection were identified upon culture of the organism by implementing the organism's unique requirements for growth. This report of the patients' clinical and immunologic course is based on a retrospective chart review. Two distinctly different presentations of M haemophilum infection were observed. Three patients presented with cutaneous lesions, typical of those seen in previous reports of the infection. Two others developed pulmonary disease only. All patients received directed therapy against M haemophilum, but respiratory failure developed in the patients with pneumonia and they died. The remaining 3 patients survived and are free of infection. These are the only reported cases of M haemophilum infection in bone marrow transplant recipients. Early diagnosis obtained through biopsy and special request for culture conditions conducive to the growth of the organism may decrease morbidity and mortality, particularly in patients with pulmonary disease. PMID- 7491701 TI - Cytomegalovirus DNA in peripheral blood leukocytes and plasma from bone marrow transplant recipients. AB - Granulocytes, monocytes, and T- and B-lymphocytes were separated from 28 blood samples collected from 5 bone marrow transplant (BMT) recipients. About 40% of granulocyte, monocyte, and B-lymphocyte samples were CMV DNA-positive by polymerase chain reaction in recipients with cytomegalovirus (CMV) infection. CMV DNA was rarely detected in separated T-lymphocytes. Within each of the simultaneously separated paired samples, there were several with single positive cell subtypes. Monocytes, granulocytes, and B-lymphocytes were the single positive samples in some instances. Thus, it is important to have all of the different cell subtypes present in samples for detection of CMV DNA in peripheral blood. We also studied the appearance of CMV DNA in plasma and peripheral blood leukocytes (PBLs) from 351 blood samples collected from 30 BMT recipients during a follow-up period of at least 3 months after BMT. All cell subtypes were represented in the PBL samples. In the 13 recipients who developed symptoms possibly associated with CMV infection or CMV disease, a correlation with the detection of CMV DNA in < or = 2 x 10(5) PBLs was found. In PBLs from 11 of the 13 BMT recipients, CMV DNA was detected before the onset of symptoms. CMV DNA was not detected in < or = 2 x 10(5) PBLs from recipients without CMV infection. The virus load in PBLs decreased during ganciclovir treatment. Nine of the 13 recipients displayed PCR-positive plasma samples, and CMV DNA was detected frequently after the onset of symptoms. PMID- 7491702 TI - Global energetic failure in brain-dead patients. AB - The objective of this prospective, clinical study of consecutive patients was to test the hypothesis of a global energetic failure in brain-dead patients by analyzing indices of peripheral oxygenation during brain-dead resuscitation. Subjects comprised 24 subjects with brain death criteria from a multidisciplinary intensive care unit. The causes of brain death were multiple: severe traumatic head injury, cerebrovascular event, cerebral anoxia, primary brain tumor, and gunshot wound to the head. Interventions used were radial and pulmonary artery catheterization. Hemodynamic and gasometric parameters and blood lactate levels were measured immediately after the diagnosis of brain death (T0) and 4 hr later (T4), while patients were receiving a therapeutic protocol (fluids, vasopressive drugs) adjusted to reach a mean arterial pressure of 75 mmHg. In 18 of our 24 patients, a blood lactate level > or = 2 mmol/L (mean +/- SD: 4 +/- 2 mmol/L) associated with an increased mean lactate to pyruvate ratio (14.4 +/- 3.2) was observed at T0, while oxygen delivery (DO2) was high (533 +/- 208 ml/min/m2) and mean arterial pressure was 76 +/- 21 mmHg. Patients were subdivided into two groups according to changes in DO2 from T0 to T4: group D comprised 14 patients (10 with hyperlactatemia and 4 with normal lactate) in whom DO2 and oxygen consumption (VO2) simultaneously decreased from T0 to T4 without significant change in lactate level; group I comprised 10 patients (8 with hyperlactatemia and 2 with normal lactate) in whom DO2 and VO2 simultaneously increased, while the blood lactate level decreased significantly from 3.5 +/- 2.5 mmol/L at T0 to 2.1 +/- 1.0 mmol/L at T4 (P < 0.05). Our results indicate that the brain-dead state was frequently associated with a global energetic failure probably due to a cellular oxygen deficit, despite blood pressure within the normal range. This energetic failure, because it is associated with high levels of DO2, could result from a defect in peripheral oxygen extraction. Aggressive therapy, achieved by producing a further increase in DO2, may reduce this global tissue oxygen deficit. PMID- 7491703 TI - Mixed allogeneic chimerism in the rat. Donor-specific transplantation tolerance without chronic rejection for primarily vascularized cardiac allografts. AB - Graft loss secondary to chronic rejection remains a major source of morbidity and mortality in solid organ transplantation. Mixed chimerism has been suggested as one potential approach to overcome this limitation. Until now, whether long-term tolerance for primarily vascularized allografts can be achieved with mixed chimerism has not been adequately assessed due to technical limitations in the mouse and the inability to establish a reliable model of mixed chimerism in the rat. We now report that stable multilineage mixed hematopoietic chimerism can be achieved following the transplantation of a mixture of T cell-depleted syngeneic and allogeneic bone marrow cells into myeloablated rat recipients using a number of MHC plus minor antigen-disparate donor and recipient strain combinations (F344+WF-->F344, F344+ACI-->F344, WF+F344-->WF, and WF+ACI-->WF). Ninety-one percent of animals engrafted with a level of lymphoid chimerism ranging between 12% and 93% (73.3 +/- 4.8%). Peripheral blood lymphocyte chimerism remained stable for up to 13 months after reconstitution. Multilineage chimerism for lymphoid (T and B cells) and myeloid (granulocyte and macrophage) lineages was present, which suggests that engraftment of the pluripotent rat stem cell had occurred. There was no clinical or histologic evidence of graft-versus-host disease. Donor-specific skin (mean survival time [MST] > or = 177 days) and primarily vascularized cardiac (MST > or = 213 days) grafts were accepted without evidence for acute or chronic rejection. In contrast, MHC-disparate third-party skin (MST = 14 days) and cardiac grafts (MST = 13 days) were rapidly rejected. The tolerance was systemic, since donor-specific tolerance was present in vitro as assessed by the mixed lymphocyte proliferation assay. These data suggest that mixed chimerism prevents graft loss secondary to chronic rejection in skin as well as primarily vascularized grafts. Furthermore, a rat model for mixed allogeneic chimerism may provide insight into the mechanisms involved in tolerance induction for a variety of allografts (lungs, small bowel, limb, etc.) not readily transplantable in mouse recipients. PMID- 7491704 TI - Gamma delta T cells play no major role in human heart allograft rejection. AB - To investigate the role of gamma delta T cells in human heart transplantation, we searched for this T cell population in endomyocardial biopsies as well as in T cell lines and clones derived from graft-infiltrating lymphocytes. The number of gamma delta T cells in endomyocardial biopsies from transplanted patients (n = 55) was mostly low and did not differ significantly from nontransplanted patients (n = 21). Moreover, there was no association of gamma delta T cell distribution with rejection status or with time posttransplantation. Graft-derived T cell lines were established in the presence of autologous feeder cells and recombinant interleukin-2 to favor the growth of in vivo-activated T cells. Twenty T cell lines analyzed by flow cytometry showed low percentages of gamma delta T cells, and we were unable to obtain gamma delta T cell clones for functional studies. These results show that gamma delta T cells are poorly expressed on human heart allograft infiltrates and indicate that, when present, they are not activated in the graft. Our data suggest that gamma delta T cells do not have a major role in human heart rejection. PMID- 7491705 TI - CD44 expression in rejecting rat small bowel allografts. AB - CD44 is a widely expressed cell surface protein that recognizes multiple ligands and is involved in extra- and intercellular adhesion. The precise role of CD44 in immune interactions is currently unknown, but it is believed to be a homing receptor involved in lymphocyte trafficking and inflammatory responses. This study investigated CD44 expression in intestinal tissue after heterotopic rat small bowel transplantation and assessed the effect of transplantation on intestinal epithelial cell proliferation using an antibody to the nuclear activation Ag Ki67. Lamina propria and intestinal epithelial cell expression of CD44 was graded blindly by five observers, and villus epithelial cells were noted as being positive or negative for Ki67 staining. CD44 expression was high in the lamina propria of both allografted and isografted animals; however, there was no significant difference between the two groups. In contrast, the expression of CD44 on the villus epithelium was greater in allografted animals and progressed toward the villus tips as rejection developed, declining thereafter because of loss of villus integrity. Ki67 positivity was also greater in allografted animals but did not progress toward the villus tip. This is the first reported observation of CD44 expression on intestinal epithelium that is not restricted to the crypts. The findings indicate the involvement of CD44 in the rejection process and demonstrate changes in the proliferative profile of rejecting small intestinal epithelium. Further studies into adhesion molecules, such as CD44, may help to improve understanding of graft failure and promote the development of new therapeutic approaches for controlling and preventing graft rejection. PMID- 7491706 TI - Differential expression of adhesion molecules during rat small bowel allograft rejection. AB - The adhesion molecules intercellular adhesion molecule 1 (ICAM-1), lymphocyte function-associated antigen-1 (LFA-1) (alpha and beta chains), and very late activation antigen-4 (VLA-4) have an essential role in cell-cell interactions and the initiation of immune responses. This study used an indirect immunoperoxidase technique to investigate the expression of these molecules in the lamina propria of allografts and isografts after heterotopic rat small bowel transplantation. Normal untransplanted small bowel served as additional controls. Overall, ICAM-1 and LFA-1 alpha expression was significantly higher in allografts, although there was variable expression of these molecules in isografted animals. There were temporal differences in the expression of ICAM-1 and LFA-1 alpha in that increased ICAM-1 expression was more pronounced in the the early posttransplant period, whereas there was a progressive increase in LFA-1 alpha as rejection developed. In contrast, there was no difference between allograft and isograft expression of LFA-1 beta and VLA-4. This study has demonstrated a preferential increase in adhesion molecule expression with developing rat small bowel allograft rejection and suggests that adhesion molecules are involved in the development and progression of allograft rejection. Although the observed differences in antigen expression are not as marked as those previously reported in other organ transplants, appropriate adhesion molecules may present suitable targets for immunotherapeutic protocols after small bowel transplantation. PMID- 7491707 TI - Inhibition of intimal hyperplasia in rat aortic allografts with cyclosporine. AB - The rat aortic transplant model of chronic rejection was used to study the effect of cyclosporine (CsA) on the development of intimal hyperplasia. ACI and Lewis rat strains were used to create isograft and allograft CsA nontreated and treated groups. After orthotopic abdominal aortic transplantation, recipients received either no treatment, CsA 2.5 mg/kg/day, CsA 5 mg/kg/day, or CsA 10 mg/kg/day by gavage. Treated grafts were harvested at 3 and 6 months after transplantation, and computer image digital analysis was used to measure intimal and medial areas of graft cross-sections. At 3 months, the reduction in percent intima was 62% (P = 0.005), 74% (P = 0.002), and 97% (P < 0.0001) for the 2.5-, 5-, and 10-mg/kg allograft groups, respectively. There was a 93% (P < 0.0001) reduction in percent intima at 6 months in the 10-mg/kg allograft group. CsA treatment also preserved the aortic media. In comparison to nontreated isografts, medial area in nontreated allografts was decreased by 37% at 3 months after transplantation. In contrast, medial area was not significantly changed in CsA-treated recipients (10 mg/kg/day) in comparison to nontreated isografts. More importantly, medial nuclear density was preserved in the CsA-treated recipients in comparison to nontreated allografts and was similar to treated or nontreated isografts. In conclusion, daily high dose CsA treatment was found to markedly inhibit intimal hyperplasia in rat aortic allografts up to 6 months after transplantation, which suggests that CsA at an adequate dosage can suppress chronic rejection. PMID- 7491708 TI - Liposomal cyclosporine. Characterization of drug incorporation and interbilayer exchange. AB - A number of previous studies have examined the application of liposomes as carriers for the immunosuppressive agent cyclosporine. These studies, however, have generated equivocal results, particularly with regard to the therapeutic properties of such systems. In the present work, we have characterized cyclosporine incorporation into well defined liposomes, large unilamellar vesicles, and have examined the stability of drug association. Contrary to some earlier reports, we show that only modest levels of cyclosporine can be accommodated in the liposomal membrane and that the extent of drug incorporation is greatly reduced as the bilayer cholesterol content is increased. Furthermore, we demonstrate that cyclosporine, despite its hydrophobic character, can rapidly exchange between vesicles. This raises the possibility that, after i.v. administration, drug migration to other blood components might negate the potential benefits arising from liposomal delivery. In a companion paper, therefore (Choice et al., Transplantation, 1995, this issue), we have followed the pharmacokinetics and biodistribution of liposomal cyclosporine in a study that examined the behavior of both the drug and the liposomal carrier. PMID- 7491709 TI - Diagnostic analysis, clinical importance and molecular basis of dihydropyrimidine dehydrogenase deficiency. PMID- 7491710 TI - Analysis of competitive agonist-antagonist interactions by nonlinear regression. AB - The rigorous estimation of a dissociation constant (Kb) for antagonists in functional assays has been sought by pharmacologists using a variety of techniques ever since the regression method of Arunlakshana and Schild in 1959. Here, Michael Lew and James Angus describe a simplified global regression method with improved accuracy compared to Schild analysis. The method is suitable for personal computers with standard graphing and statistical software. The accuracy of the predicted pKb values and confidence intervals has been tested by comparing examples of published data, and by mathematical (bootstrap) simulations. PMID- 7491711 TI - Functional consequences of retrograde axonal transport of receptor-bound neurotensin. AB - Transport mechanisms or trafficking systems in axons allow neuronal constituents to reach nerve terminals. Accordingly, presynaptic receptors undergo axonal transport through a process that is fast and bidirectional. However, axonal transport of receptors also appears to be involved in the 'flow' of information from membrane receptors to genome. The most compelling evidence that retrograde axonal transport of receptor-bound neuropeptide represents a long-distance signalling system in gene expression, discussed here by Pierre Laduron, comes from the recent observation that neurotensin injected in the striatum undergoes retrograde transport in dopaminergic neurones to the substantia nigra, where it induces in the cell bodies an increase of tyrosine hydroxylase mRNA. Although the molecular mechanisms of this remain to be elucidated, the present data suggest the existence of retrograde signal molecules, or 'third messengers', which have to be associated with synaptic or endocytotic vesicles in order to gain entry into the cell bodies, where they may modulate gene expression. PMID- 7491712 TI - Mapping of opioid receptors using antisense oligodeoxynucleotides: correlating their molecular biology and pharmacology. AB - Understanding the pharmacology of opioid receptors took a major step forward with the cloning of genes that encode four members of the opioid receptor family. Gavril Pasternak and Kelly Standifer show how strategies that use antisense oligodeoxynucleotides can provide a selective approach to correlate the properties of the cloned receptors with their in vivo pharmacological effects, and have confirmed the association of the delta-, mu- and kappa 1-opioid receptor clones with analgesia mediated through delta-, mu- and kappa 1-opioid receptors, respectively. Approaches that use antisense oligodeoxynucleotides provide an opportunity to characterize the pharmacology of short cDNA fragments without necessitating the cloning of the entire cDNA and can assess the role of the products of specific exons in a receptor, providing an approach for the study of alternative splicing as demonstrated with the mu- and kappa 3-opioid receptor clones. PMID- 7491713 TI - Delayed protection of the heart against ischaemia. AB - Ischaemic preconditioning is the protective adaptive mechanism produced by short periods of ischaemic stress that results in a marked resistance of the myocardium to prolonged periods of the same stress; however, this protection is transient. There is now evidence that protection resulting from preconditioning returns several hours later, and here James Parratt and Laszlo Szekeres highlight the possible importance of this concept, which may lead to novel approaches to the long-term protection of the heart against ischaemic injury. PMID- 7491715 TI - [Treatment of patients and safety--significance of the Cochrane databases]. PMID- 7491714 TI - NMDA receptor-dependent excitotoxicity: the role of intracellular Ca2+ release. AB - Massive activation of glutamate receptors can result in excessive rises in cytoplasmic Ca2+ that are thought to underlie the fundamental processes ultimately leading to neuronal death. Preventing such cellular Ca2+ rises in the brain may reduce considerably the neuronal damage produced by stroke, head trauma, or epilepsy. Activation of NMDA receptors is instrumental in this type of neurotoxicity. Recent findings, discussed here by Istvan Mody and John MacDonald, indicate that a large proportion of the neurotoxic Ca2+ that enters nerve cells following NMDA receptor activation originates from an intracellular Ca2+ pool. The release of Ca2+ from this pool is sensitive to the skeletal muscle relaxant dantrolene, and this may constitute a novel and alternative therapeutic approach against NMDA receptor-mediated excitotoxicity. PMID- 7491716 TI - [Postmyelographic headache]. AB - Prolonged cerebrospinal fluid leakage from the dural puncture site is considered the most important factor in the development of post-myelography headache. Incidences of post-myelography headache and post dural puncture headache vary a lot--possible reasons are discussed. The statement that patients with a low substance P level in cerebrospinal fluid are prone to developing a post dural puncture headache seems especially interesting. Articles considering clinical as well as experimental studies are reviewed. PMID- 7491717 TI - [Cognitive symptoms in persons with snoring and sleep apnea. An epidemiologic study of 1.504 women and men aged 30-60 years. The Dan-MONICA II study]. AB - The objective of the study was to evaluate the relation between every-night (habitual) snoring, sleep apnoea and cognitive complaints (concentration and memory complaints) in an adult population-based sample. In the DAN-MONICA (MONItoring trends in CArdiovascular iseases) 1504 males and females aged 30, 40, 50 and 60 years were classified according to their snoring habits. Nocturnal respiration was measured in 748 participants. The following measures were regarded as potential confounders: age, gender, unintended sleepiness, insomnia, depression, hypnotic use, alcohol consumption by questionnaire, body mass index (BMI) and blood pressure. Concentration and memory problems were both related to depression, insomnia and unintended sleepiness. Snoring and sleep apnoea (defined as a respiratory distress index-RDI > or = 5) were associated with concentration problems and unintended sleepiness. The odds ratios (95% confidence intervals) between snoring, concentration and memory problems, calculated by logistic regression analysis after adjustments for the above confounders, were 1.90 (1.23 2.91, p < 0.01) and 1.38 (0.97-1.99, NS). For those with sleep apnoea, the odds ratios were 3.53 (1.42-8.73. p < 0.001) and 1.51 (0.81-2.14, NS) for concentration and memory problems, respectively. The main conclusion drawn from this study is that cognitive complaints show a high correlation to mood, insomnia and hypersomnia. Habitual snoring and sleep apnoea show a correlation to concentration problems, but not to memory problems. This suggests that part of the association between snoring, sleep apnoea and cognitive dysfunction is related to the presence of sleep disturbances and daytime sleepiness. PMID- 7491718 TI - [Intravascular ultrasound and percutaneous transluminal angioplasty. Preliminary results]. AB - Intravascular ultrasound is a new imaging modality which enables cross-sectional images of blood vessels. These preliminary results of intravascular ultrasound used before and after percutaneous transluminal angioplasty (PTA) in 19 patients, confirm earlier reports of the ability to distinguish between the three different layers of the vessel wall. Atherosclerotic plaques may also be assessed as well as a quantification of the stenosis. After PTA changes in the plaques such as intimal fractures, flaps and dissections could be recognized, and the effect of the intervention could be quantified by measuring the reduction of stenosis and increment of the luminal area. The increased luminal area following PTA was partly due to stretching of the artery and partly due to compression of the atheroma. We found a correlation between the amount of calcium in the plaque and the size of the dissection after PTA. Comparing IVUS with angiography revealed an acceptable agreement in quantifying stenoses before the intervention but not after. We conclude that IVUS is a feasible method to evaluate stenosis in blood vessels before and after PTA. By supplying additional information to angiography, it may improve the selection and outcome after PTA. PMID- 7491719 TI - [Community psychiatry in Copenhagen. Services and resource utilization]. AB - The purpose was to estimate the number and nature of contacts in a newly established community psychiatry centre and their distribution according to diagnostic category, registered on computer. The number of contacts and day patient days were about 6000 each per year. Two professionals participated in 20% of the contacts and 20% were in the patient's home (psychotic patients). Fifty percent of the contacts had a psychotherapeutic content, 10% were about medicine and 30% had a mixed content. The chronic patients made 20-25 contacts/year and other patients 6-10 per treatment episode. Forty-five percent of the resources were used on out-patients/hometreatment, 30% on activities, 10% on intake, 10% on secretarial work and 5% on administration. Seventy percent of resources were used on chronic patients and 20% on patients with minor disorders. Fifty-five percent of time spent was used for direct patient contact (40% for psychiatrists). Time for contacts was 45-60 minutes/patient/2 weeks (6 minutes for psychiatrists). In accordance with the politically defined goal the main part of the resources were used for psychotic patients. The patients valued the contacts as seen by a low rate of non attendance. The amount of time for contact seems reasonable although it was low for psychiatrists. PMID- 7491721 TI - [Should fasting prior to ultrasound examination of the gallbladder, the biliary ducts and the pancreas have to include not-drinking?]. AB - From May to September 1991, 49 patients were examined by ultrasound (Picker CS 9000, 3.5 MHz curved array transducer) before and 30 minutes after drinking 150 ml of water. The volume of the gallbladder decreased significantly from a mean of 23.4 ml to 20.1 ml (p = 0.04). No significant change was found either between the visualisation of the gallbladder and bileducts (0.20 > p > 0.10) or the visualisation of the pancreas (0.40 > p > 0.30). However in 10 patients with cholelithiasis, the calculi were only seen in eight patients after drinking water. Thus, it must still be recommended that patients neither eat nor drink for four hours prior to an ultrasound examination of the gallbladder. PMID- 7491720 TI - [Shore-term stimulation with myeloid growth factors expands bone marrow hematopoiesis. A magnetic resonance spectroscopic study]. AB - The present study is intended to investigate the expansion of haematopoiesis by localised volume selective proton magnetic resonance spectroscopy (MRS) during treatment with myeloid growth factors. Six consecutive patients were treated with daily subcutaneous injections of recombinant human granulocyte colony-stimulating factor (rhG-CSF, n = 2) or granulocyte-macrophage colony-stimulating factor (rhGM CSF, n = 4) for five days before marrow harvest. MRS investigations were performed prior to treatment (day 0), day 5 and day 12. The patients responded with a rise in blood absolute neutrophil count from median 3.3 x 10(9)/l (range 1.3-7.3 x 10(9)/l) before to 15.6 x 10(9)l (range 6.8-22.0 x 10(9)/l) after treatment. Concomitantly an increase in bone marrow cellularity and myeloid:erythroid ratios documented the stimulation of myelopoiesis. During priming, the light-density cell proliferation rate in marrow samples increased from median 21.9 (range 4.5-31) x 10(3) cpm to 54.7 (range 13.9-94) x 10(3) cpm and the total number of myeloid progenitors enumerated as day 7/14 GM-CFUs per volume aspirated marrow increased from median 11/8 x 10(3) (range 4.0-87.5/2.2 103.0) to 64/76. x 10(3) (range 28.4-1180.6/23.2-2850.0). MRS detected a significant increase in bone marrow "relative water content" day 12, one week after myeloid growth factor treatment was stopped, from median 30.5% (range 16 45) to 79% (range 56-93) (p < 0.05). Haematopoiesis was concommittantly detected in new areas of femur.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7491722 TI - [Respiratory insufficiency caused by a vocal cord polyp. An accidental finding during respiratory function tests]. AB - We present a case in which a 58-year-old woman was admitted for spirometry due to progressive dyspnoea. She had a history of chronic obstructive pulmonary disease treated for more than 20 years. The flow-volume loop indicated an upper airway obstruction. An otolaryngologic examination revealed a vocal cord polyp. It was excised and the patient's dyspnoea was relieved. The following spirometry indicated that the upper airway obstruction had been removed. The aim of this case report is to demonstrate that the flow-volume loop is a useful method in diagnosing and quantifying upper airway obstructions. PMID- 7491723 TI - [Chronic lymphocytic and myeloid leukemia in the same patient]. AB - A patient with chronic lymphocytic leukemia (CLL) treated with chlorambucil and prednisone developed chronic myelocytic leukemia (CML) five years later. An association between the two chronic leukemias has only rarely been reported. In the majority of cases the CLL preceded CML after which the malignancies coexisted morphologically in bone marrow and peripheral blood. In this case we report a suppression of CLL related symptoms and blood findings after the emergence of CML. However, immunophenotypic studies demonstrated a minor remaining population of CLL cells in the peripheral blood. PMID- 7491724 TI - [Helicobacter pylori status--how, when and in whom?]. PMID- 7491726 TI - [Serotonin uptake inhibitors and adverse effects]. PMID- 7491727 TI - [CT and MR scanning as a part of the "wait and scan" policy in acoustic neurinoma]. PMID- 7491725 TI - [How are the bladder tumors registered actually?]. PMID- 7491728 TI - [Size of acoustic neurinomas assessed by CT and magnetic resonance scanning]. AB - Some acoustic neuromas do not grow or grow very slowly. It is therefore of value to compare the results of measuring the size of acoustic neuromas by either CT or Magnetic Resonance Imaging (MRI) scanning with a view to being able to observe the growth rate of these tumours. Fifteen patients with acoustic neuromas had both CT- and MRI-scans performed. Two trained radiologists evaluated the size of the tumours in a blinded fashion. There was a significant difference between one observer's calculations from the CT- and the MRI-scans, and also a significant difference between the two observers' calculations from the CT-scans. No difference in calculating tumour size was found between the two observers' calculations from the MRI-scans. It is concluded that the size of acoustic neuromas i evaluated more equally by two observers using MRI-scans than when using CT-scans. By the use of repeated MRI-scanning it is possible to keep patients with acoustic nouromas under observation and avoid operation in those patients where the tumour does not grow. PMID- 7491729 TI - [Tuberculosis in low prevalence countries. Back to the future?]. AB - The incidence of tuberculosis has declined in all European countries in this century but in several countries the decline has come to a stop. Microscopy a.m. Ziehl-Neelsen and culture are still the mainstays in the diagnosis of tuberculosis but new DNA-technics such as PCR and RFLP are increasingly used. The tuberculin test is hitherto the only practically feasible test for identification of subjects infected with M. tuberculosis. BCG is the most used vaccine in the world but the protective mechanisms are not fully understood and the efficacy controversial. Immunotherapy with M. vaccae may be used as an adjuvant to chemotherapy. Few cases of tuberculosis are observed in AIDS-patients in Denmark. Modern standard treatment of tuberculosis comprises a six-month regimen with isoniazid, rifampicin, pyrazinamide and possibly ethambutol. Multidrug resistance in tuberculosis is rare in Western Europe. Chemoprophylaxis with isoniazid is debatable. Patients with smear-positive sputum may be infectious whereas patients with extrapulmonary tuberculosis normally pose no risk. PMID- 7491730 TI - [Pharmacological routes of administration in circulatory collapse]. AB - A review of different ways of injecting drugs during cardiopulmonary resuscitation (CPR) and during states of shock is presented. On the basis of this review, central or peripheral intravenous injection is recommended as first choice. If a peripheral vein is used, the drug injection should be followed by infusion of a large volume of normal saline to facilitate the entry of the drug into the central circulation. In case of endotracheal intubation, several drugs can be injected into the tracheal tube. Atropine, lidocaine and naloxone are shown to be effective when given by this route. Adrenalin is probably not effective when injected endotracheally. Intraosseous injection of drugs, crystalloids and colloids is an alternative in all states except for hypovolaemic shock due to lower infusion rates. Intracardiac injection of drugs during CPR is recommended as a last resort. PMID- 7491731 TI - [Late symptoms among patients surgically treated for breast cancer. A questionnaire in the county of Southern Jutland]. AB - The 1989 protocol of the Danish Breast Cancer Cooperative Group implied a more extensive axillary dissection and reduced frequency of regional lymphnode irradiation. A questionnaire concerning late symptoms was sent to all women operated from 1989 to 1993. The rate of response among women treated with modified radical mastectomy was 82%. Of 163 patients, 79% had one or more late symptoms. Most troublesome was pain in the neck, shoulder and arm or scar-related pain. Other symptoms were lymphoedema, paraesthesia of the arm and impaired shoulder function; each symptom occurring among 29-47% of the women. Most symptoms began three to four weeks postoperatively. The frequency of late symptoms corresponded with number of lymphnodes removed, number of tumourpositive nodes and irradiation of the scar. Thirty-five women were treated with tumourectomy and they experienced similar late symptoms with a similar degree of distress. It is concluded that late symptoms among women treated for breast cancer still occur frequently. PMID- 7491732 TI - [Acute pulmonary infections at autopsy. A study of clinical and macroscopic diagnoses at autopsy compared with microscopic autopsy findings]. AB - The clinical diagnoses and gross diagnoses of acute pulmonary infections were investigated for 100 consecutively performed hospital autopsies, and the diagnoses were compared with the histological findings. We found 34 cases of pneumonia and one case of tuberculosis. Of these, 29 infections represented principal diagnoses, i.e. as causative of or contributive towards death. The predictive values, the sensitivities and specificities were estimated. The predictive value for gross diagnostics was low with a value of 42.9% against 58.8% for clinical diagnostics. The sensitivities for both clinical diagnostics and gross diagnostics were about 30% and the specificities about 85%. The study shows that sampling for histology from all pulmonary lobes is essential for correct autopsy diagnoses, either from areas that appear to be infected on gross examination or from the peripheral parts. Furthermore autopsy performance is still of great value for clinical diagnostics and for medical statistics. PMID- 7491733 TI - [Body proportions of Danish children. Curves for sitting height ratio, subischial length and arm span]. AB - No previous Danish growth charts have been published for sitting height, subischial leg length, armspan or the ratio of sitting height to stature. These growth charts are useful in the evaluation of dysproportional growth retardation. Foreign standards have previously been used. However, due to population-specific differences and "the secular trend", the construction of Danish growth charts was found to be appropriate. We examined 1977 children (927 boys and 1050 girls) from the Copenhagen area, aged six to 20 years. Height, sitting height and armspan were measured. Growth charts were constructed using local linear regression and smoothing of residuals. We found sitting height values in Danish children to be comparable to English and Norwegian children. Sitting height values were lower than values of Dutch children, but higher than values of Swiss and Swedish children. The extremities of Danish children were 3-4 cm longer than the extremities of English children, but Dutch children had 2-3 cm longer armspan than the Danish boys and girls. Growth charts for sitting height, sitting height ratio, subischial leg length and armspan are presented. These charts should be useful when evaluating dysproportionality in growth retarded children and when assessing the effect of growth modulating therapy in children. PMID- 7491734 TI - [Adnexial torsion caused by ovarian hyperstimulation syndrome]. AB - A case of adnexal torsion of an enlarged ovary during early pregnancy after gonadotropin therapy is presented. Adnexal torsion is a rare but serious complication to ovarian hyperstimulation. The diagnosis is difficult, but the combination of ovarian enlargement, abdominal pain, nausea, progressive leukocytosis and anaemia may indicate torsion of the adnex. Prompt diagnosis is important. Simple unwinding of the twisted adnex by a laparoscopic technique has been successful in most cases. PMID- 7491736 TI - [Effectiveness of vaccination against influenza in the elderly]. PMID- 7491735 TI - [Post-traumatic pseudoaneurysm of the posterior tibial artery]. AB - A case of post-traumatic pseudoaneurysm of the posterior tibial artery is presented. The diagnosis was made by ultrasound one week after the trauma, and a vascular reconstruction was performed. The purpose of this case report is to bring attention to this complication to vascular trauma, which may cause delayed symptoms long time after the injury. We suggest that these patients should be treated with vascular reconstruction rather than ligation, thereby preserving the function of this artery. PMID- 7491737 TI - [Does the National Board of Health adn Welfare support worthless research on alternative therapy?]. PMID- 7491738 TI - [Status of the psychotherapy in general practice?]. PMID- 7491739 TI - Ultrasonic approach to obtaining partial thermodynamic characteristics of solutions. AB - We describe a method for evaluating the thermodynamic characteristics both of pure liquids and of solutes in solutions using data derived from ultrasonic velocity measurements. The principal possibility of using ultrasound velocity lies in the fact that the velocity of ultrasound is a simple function of the adiabatic compressibility. The problem is formulated as an initial value problem for the parabolic type differential equations in partial derivatives. The validity of the method is demonstrated by calculation of the thermodynamic parameters for water, glycine and alanine in aqueous solutions at infinite dilution. PMID- 7491740 TI - Tissue characterization of arteries with 4 MHz ultrasound. AB - Reflections of 4 MHz ultrasound pulses from 60 sites on 15 human aorta specimens were analysed using the plane-wave Born approximation (PWBA) inverse scattering method. The recovered high-resolution acoustic impedance profile for each site was correlated with an analysis (by microscope) of sections taken from that site. The shape of the impedance profile was found to be sensitive to the pathology of the tissue at each site. The severity of the various conditions could be gauged from the recovered impedance profiles. Some of the insonifications were done on aorta specimens that were shielded from the ultrasound transducer by a human tissue specimen about 1.25 cm thick in order to study the effects of intervening tissue. We found that it was possible to compensate for the distortion due to the frequency dependent attenuation of the intervening tissue. We conclude that ultrasound imaging, with the reflections analysed by means of PWBA inverse scattering provides a promising non-invasive tissue characterization modality. PMID- 7491741 TI - Ultrasonic tissue characterization of end-stage dilated cardiomyopathy. AB - The clinical use of angiotensin-converting enzyme (ACE) inhibitors now represents the standard of care for the treatment of chronic congestive heart failure. We utilized ultrasonic tissue characterization to define potentially salutary effects of treatment with ACE inhibitors on the material properties of the heart and its potential influence on cardiac remodeling at the cellular level. Ten 1 month-old cardiomyopathic (CM) Syrian hamsters were treated with captopril (2 g/L water ad libitum), and 10 CM and 5 normal hamsters were maintained untreated for 9 months. Hearts were excised, and backscattered radio-frequency data were acquired from 1200 independent sites from each specimen with a high-resolution 50 MHz acoustic microscope for calculation of integrated backscatter. Treatment with captopril elicited relative decreases in left ventricular (LV) wet weight, LV calcium concentration and integrated backscatter (IB) in treated as compared with untreated cardiomyopathic hearts without affecting LV collagen concentration. The IB from hearts of treated cardiomyopathic hamsters was significantly less in both grossly normal regions of myocardium (P < 0.02) and scar tissue regions (P = 0.0005) as compared with IB from hearts of untreated hamsters. The reduced integrated backscatter from hearts of treated cardiomyopathic hamsters indicates direct alterations in the material properties of cardiomyopathic hearts after captopril therapy. The lower IB from scar tissue in the treated animals was associated with decreased scar tissue calcification, which represents a novel therapeutic effect of captopril. This is the first report that delineates direct effects of ACE inhibitors on the material properties of both scar tissue and grossly normal myocardium at the cellular level in experimental dilated cardiomyopathy.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7491742 TI - Assessment of the fetal PO2 changes by cerebral and umbilical Doppler on lamb fetuses during acute hypoxia. AB - The objective of the present study was to validate one or a combination of fetal Doppler parameters in order to assess acute fetal hypoxia in an ovine model. Acute hypoxia was induced by reducing umbilical, or maternal aortic flow (approx. 70%). A CW Doppler probe was fixed on the fetal cervical skin, facing the internal carotid artery and the fetal abdominal skin adjacent to the umbilical arteries. (The angle between Doppler beam and flow vector remained constant.) A "Doptek 3000" spectrum analyser was used to measure the maximal and mean Doppler frequencies. Heart rate (HR), umbilical blood flow (UBF), carotid blood flow (CBF), umbilical RI (URI), cerebral RI (CRI) and cerebroplacental ratio (CPR = CRI/URI) were calculated in real time. A catheter was inserted into the fetal femoral artery, for blood gas (PO2, PCO2 pH) and blood pressure (BP) measurements. After 1 min of aorta compression (70% aortic flow reduction), the URI increased by 10% (P < 0.05), and the UBF decreased by 10% (P < 0.05), but the CRI decreased by 20% (P < 0.02), and the CBF did not change significantly. Fetal PO2 and CPR fell down after 1 min (59% and 38%, respectively; P < 0.001), although strong fetal heart rate decelerations were observed. The blood pressure, PCO2 and pH did not change significantly during this test. Throughout the 12 min of cord compression (70% umbilical flow reduction) the URI increased (70% to 80% P < 0.001), and the UBF decreased (approx. 60%; P < 0.001), but the CRI decreased (approx. 25%; P < 0.01), and the CBF remained constant (+/- 5%; ns). Fetal PO2 and CPR all decreased during the compression (30% to 44% and 40% to 60%, respectively; P < 0.001). HR, pH and PCO2 did not change significantly. During cord compression the blood pressure did not change significantly. In both cases, the CPR decreased significantly (P < 0.001) with the PO2 in the same direction and with a comparable amplitude (-30% to -50%). Nevertheless, the drop in CPR was greater during cord compression than during aorta compression, probably because the compression of the cord induced a central hypovolemia in addition to the hypoxia. The CPR was found to be the hemodynamic parameter that followed most closely the PO2 acute changes. The amplitude of the variations of this parameter (-30% to -50%) were quite similar to those of the PO2 during the period of acute hypoxia.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 7491744 TI - Experimental velocity profiles and volumetric flow via two-dimensional speckle tracking. AB - The performance of a two-dimensional speckle tracking system in measuring in vitro laminar flow is evaluated. The system uses a pattern matching algorithm to track subresolution-sized speckle regions between successive ultrasonic 2D pulse echo acquisitions in order to determine both the axial and the lateral components of velocity. In this study, multiple 2D vector velocity maps were acquired in real time using a calibrated laminar flow phantom, and then statistically analyzed off-line. At a 90 degrees transducer angle, volumetric flow rates computed from measured velocity profiles exhibited excellent linearity (R2 > 0.99), with a mean error of -6.1%, over the range 5-30 mL/s. At 105 degrees and 120 degrees, experimental volume flow rates also agreed well with actual rates, although measured velocity profiles appeared more irregular with decreasing Doppler angles. Velocity profiles estimated using sampled radio-frequency data rather than envelope-detected data were inconsistent due to an insufficient sampling rate and the quantization of the velocity grid. Results indicate that excellent flow velocity and volume rate estimates can be obtained from vector velocity measurements along a single line of sight, without a priori knowledge of the flow direction, at transducer angles near 90 degrees where Doppler instruments are prone to large errors. PMID- 7491743 TI - Elasticity imaging for early detection of renal pathology. AB - Early detection of renal pathology may be possible with elasticity imaging. This hypothesis was experimentally tested by quantitatively imaging internal mechanical strain due to surface deformations in an in vitro animal model of nephritis. Preliminary data support the hypothesis that kidney elasticity changes with renal damage and concomitant scarring before problems are detectable by traditional diagnostic techniques such as laboratory measurements of renal function. PMID- 7491745 TI - A model based on dimensional analysis for noninvasive quantification of valvular regurgitation under confined and impinging conditions: in vitro pulsatile flow validation. AB - A technique is proposed for the noninvasive quantification of regurgitant flows under confined and impinging conditions. Its use requires only the knowledge of the jet orifice velocity, receiving chamber diameter, orifice-to-end wall distance and any downstream jet centerline velocity at a known distance from the orifice. The technique is based on dimensional analysis and provides a prediction of peak regurgitant flow rates. To validate the technique, known physiologic pulsatile flows were pumped through 2- and 4-mm circular orifices at 70 to 150 beats/min, into two different receiving chambers of 51 and 88 mm in diameter. At each heart rate, the peak orifice velocity was varied from 2 to 5 m/s, and the orifice-to-end wall distance was varied from 30 to 93 mm. Centerline velocities were recorded by pulsed Doppler ultrasound and averaged over multiple beats. A dimensional analysis of the parameters of the study provided an equation relating normalized centerline velocity to orifice-to-end wall distance, chamber diameter and downstream location. Statistical modeling of the experimental data was performed to compute the constants involved in this equation. The estimated (i.e., predicted by the technique) peak regurgitant flow rates were found to fall within 10% of the actual values, when centerline velocities were measured over a range of centerline distances from six orifice diameters to 85% of the chamber length. Therefore, the proposed technique provides, for the first time, a quantitative method for calculating valvular regurgitant flow rates under confined and impinging conditions. PMID- 7491746 TI - Intravascular ultrasound image subtraction: a contrast enhancing technique to facilitate automatic three-dimensional visualization of the arterial lumen. AB - At 30 MHz, the intravascular ultrasound backscatter of blood confounds the discrimination of the lumen from the arterial wall. This study validates a subtraction method which creates a still-frame image with a sharp demarcation of the lumen. The method involves subtraction of consecutive images and 2D ensemble averaging of the absolute pixel values. Subtraction exploits the dynamic properties of flowing red blood cells. Three phantom arteries were used, with erythrocytes in their lumens and wall. For this reason, it was not possible, in one single original image, to discriminate the blood in the lumen from the phantom wall. Based on 26 consecutive original images, in the mean subtraction image contrast between lumen and phantom wall grey values increased eightfold from 10.9 (5.3-19.2) (mean and range) in the original image to 87.7 (73.6-107.0) (P < 0.001). A sufficiently large contrast increase to allow automatic segmentation was obtained by using five original images (0.3-s acquisition time) for any single mean subtraction image. Low blood flow velocities (down to 0.5 cm/s) did not alter this result. Automatic segmentation of the lumen allowed fast 3D reconstruction of the lumen in all three phantom arteries. In phantom arteries, the intravascular ultrasound image subtraction technique improved contrast between lumen and wall which enabled automated lumen segmentation and fast 3D visualization of both the lumen and defects in the wall. PMID- 7491747 TI - Statistical bias and variance in blood flow estimation by spectral analysis of Doppler signals. AB - The stochastic nature of the Doppler signal is discussed as a source of variability and bias in estimation of mean blood velocity and flow performed using the Discrete or Fast-Fourier Transform. The estimators studied are those appropriate when the ultrasound beam is assumed to be wide enough to insonate the cross-section of the vessel uniformly, and assumed to be narrow enough to insonate only a diameter. Approximate expressions are derived theoretically for the biases and variances of these statistics when the Doppler power spectrum is uniform. For more complex spectra numerical evaluation is carried out by computer. Results for the double-sided spectra obtained from mixed flow are inferred from the single-sided cases. Typically, an estimate of instantaneous mean velocity has very little bias added (< 1%) but might have a standard error of approximately 10% of its mean value, and an estimate of flow in one cardiac cycle a standard error of approximately 1% of its mean value. PMID- 7491748 TI - Velocity matched spectrum analysis: a new method for suppressing velocity ambiguity in pulsed-wave Doppler. AB - A new approach to spectrum analysis, which is capable of suppressing velocity ambiguity in pulsed-wave ultrasonic Doppler, is presented. By simultaneous processing of several data samples from a range in depth, the movement of the scatterers along the ultrasonic beam can be tracked from pulse to pulse for each velocity component in the spectrum. In this way the correlation length of the signal component arising from a specific velocity increases when that velocity matches the expected velocity. The resulting velocity/time spectral display shows a more clearly defined spectral envelope of the maximum velocity than with conventional methods based on the discrete Fourier transform of the Doppler signal. This makes it possible to delineate velocity waveforms with peak velocity up to several times the Nyquist limit. Experimental data from human subclavian and aortic arteries are presented, where the new method is compared to conventional spectrum analysis. PMID- 7491749 TI - The effect of echo suppression on the mean velocity estimation range of the RF cross-correlation model estimator. AB - Ultrasound systems are widely used to visualize in real-time internal structures and blood flow velocity distributions. The latter are estimated from the received ultrasound radio frequency (RF) signals using a mean frequency estimator, capable of estimating the temporal and spatial mean frequency within a given depth and time window (estimation window). Since ultrasound RF-signals are composed of three major signal components (reflections, scattering and noise) it is necessary to suppress the power of the reflections or reverberations (stationary echo removal) to estimate the temporal mean frequency of the signal component induced by scattering originating from red blood cells. However, the stationary echo removal filter in front of the mean frequency estimator will restrict the temporal mean frequency estimation range because the power of the signal component induced by the slowly moving blood is suppressed as well. This article describes a stationary echo removal filter, acting in the RF-domain and adaptive to the temporal mean frequency of the signal component induced by stationary or slowly moving reflections and reverberations. Furthermore, a comparison in performance of this adaptive stationary echo removal filter and several static stationary echo removal filters is presented. For direct comparison of these filter algorithms as far as the effect on the temporal mean frequency estimation range is concerned, the same signal conditions in combination with the same RF domain mean frequency estimator were used in this comparison. It could be demonstrated that the adaptive echo removal filter, in combination with the RF domain mean frequency estimator used, exhibits the best performance. PMID- 7491750 TI - Physical factors involved in stress-wave-induced cell injury: the effect of stress gradient. AB - We have studied the biological effects of ablation-induced stress waves in vitro. Mouse breast sarcoma cells (EMT-6) were exposed to stress waves that differed only in rise time. Two assays were used to determine cell injury: incorporation of tritiated thymidine (viability assay), and transmission electron microscopy (morphology assay). We present evidence that the rise time of stress waves can significantly modify cell viability and that cell injury correlates better with the stress gradient than peak stress. PMID- 7491751 TI - Histologic effects of high intensity pulsed ultrasound exposure with subharmonic emission in rabbit brain in vivo. AB - In this study, the threshold for subharmonic emission during in vivo sonication of rabbit brain was investigated. In addition, the histologic effects of pulsed sonication above this threshold were studied. Two spherically curved focused ultrasound transducers with a diameter of 80 mm and a radius of curvature of 70 mm were used in the sonications. The operating frequencies of the transducers were 0.936 and 1.72 MHz. The sonication duration was varied between 0.001 and 1 s and the repetition frequency between 0.1 and 5 Hz. The threshold for subharmonic emission at the frequency of 0.936 MHz was found to be approximately 2000 W cm-2 and 3600 W cm-2 for pulse durations of 1 s and 0.001 s, respectively. The threshold was approximately 1.5-fold as high at a frequency of 1.72 MHz. However, there was considerable variation from experiment to experiment. The multiple pulse experiments at a frequency of 1.72 MHz and an intensity of 7000 W cm-2 showed that the histologic effects ranged from no observable damage of the tissue, to blood-brain barrier breakage, to local haemorrhagia, to local destruction of the tissue, to gross hemorrhage resulting in the death of the animal. The severity of the tissue damage increased as the pulse duration, number of pulses and their repetition frequency increased. The results indicate that the end point of the tissue damage may be controlled by selecting the sonication parameters. Such control over tissue effects can have several different applications when brain disorders are treated. PMID- 7491752 TI - Microscopic lung alterations and reduction of respiration rate in insonated anesthetized swine. PMID- 7491753 TI - Transcriptional regulation: lessons from the human neurotropic polyomavirus, JCV. PMID- 7491754 TI - Myristylation of the hepatitis B virus large surface protein is essential for viral infectivity. AB - The hepatitis B virus (HBV) envelope contains equimolar amounts of three viral proteins: the major (S), middle, and large (L) polypeptides. Their roles in the adsorption and penetration of the virus have not yet been elucidated. We have used a highly efficient in vitro model that permits reproducible HBV infection to investigate whether N-myristylation, a posttranslational modification of the L protein, was essential for viral infectivity. A point mutation abolishing myristylation was introduced into the HBV genome. Mutant virions were produced by transfecting viral DNA into hepatoma cells and their infectivity was evaluated on primary human hepatocyte cultures. No difference between mutant and wild-type viral RNA production could be observed. Furthermore, intermediate DNA replicative forms were observed in transfected cells demonstrating replication competence of mutant viral genomes. In addition, complete virions were produced in the cell supernatant. However, we found that mutant viral particles contained viral DNA with a reduced mean size, probably corresponding to a larger single-stranded region in the relaxed circular DNA form. We have evidenced the presence of pre S1, pre-S2, and S epitopes at the outer surface of these virions by using immunoprecipitation with specific monoclonal antibodies. This result confirmed that mutant viruses were normally assembled. By contrast, myristylation-defective mutants completely lost their infectivity for human hepatocytes in primary cultures as shown by the absence of HBs antigen production and viral intermediate replicative forms in hepatocytes. In conclusion, the myristylation of the L protein is not required for the production of Dane-like particles but it is absolutely necessary for HBV infectivity. PMID- 7491755 TI - Gene 2 of the sigma rhabdovirus genome encodes the P protein, and gene 3 encodes a protein related to the reverse transcriptase of retroelements. AB - The nucleotide sequence of the genes 2 and 3 of the Drosophila rhabdovirus sigma was determined from cDNAs to viral genome and poly(A)+ mRNAs. Gene 2 comprises 1032 nucleotides and contains a long ORF encoding a molecular weight 35,208 polypeptide present in infected cells and in virions which migrates in SDS-PAGE as a doublet of M(r) about 60 kDa. The distribution of acidic charges as well as the electrophoretic properties of the protein are characteristic of the rhabdovirus P proteins. Gene 3 comprises 923 nucleotides and contains a long ORF capable of coding a polypeptide of 298 amino acids of MW 33,790. The putative protein (PP3) is similar in size to a minor component of the virions. Computer analysis shows that the sequence of PP3 contains three motifs related to the conserved motifs of reverse transcriptases. PMID- 7491758 TI - The replication in vitro of the gammaherpesvirus bovine herpesvirus 4 is restricted by its DNA synthesis dependence on the S phase of the cell cycle. AB - Because several observations have suggested that replication of the gammaherpesvirus bovine herpesvirus 4 (BHV-4) is influenced by the physiological state of the host cell, a study was carried out to determine the relationship between BHV-4 infection and the cell cycle. The temporal expression of BHV-4 late (L) proteins in unsynchronized cell cultures was first investigated by flow cytometry. Interestingly, L protein expression occurred in a limited number of cells infected with a high multiplicity of infection, and a reciprocal correlation between the percentage of positive cells and the cell density at the time of infection was demonstrated. Moreover, the finding that a BHV-4 early-late protein was expressed in nearly all the cells suggested that a blockage in the viral replication cycle occurred in some infected cells at the stage of viral DNA synthesis or L protein expression. Because this blockage could be the consequence of the dependence of one or both of these events on the cell cycle, they were investigated after infection of synchronized cell cultures. The following findings were made. (i) Cell transition through the S phase quantitatively increased the rate of BHV-4 DNA replication. (ii) BHV-4 DNA synthesis could not be detected in cells arrested in G0. (iii) Synchronization of MDBK cells with Lovastatin before infection increased the percentage of cells expressing L proteins. (iv) In contrast, infection of cells arrested in G0 led to few positive cells. Taken together these results showed that BHV-4 DNA replication and consequently the expression of L proteins are dependent on the S phase of the cell cycle. This dependence could be of importance for several biological properties of BHV-4 infection in vitro and might have implications for the biology of the virus in vivo. PMID- 7491757 TI - Sequences required for the nuclear targeting and accumulation of human papillomavirus type 6B L2 protein. AB - The L2 protein of the human papillomaviruses is a minor structural protein and is not necessary for the major L1 protein to assemble into capsids. However L2 protein binds DNA and enhances the efficiency of HPV capsid assembly, suggesting an important role in the production of infectious papillomaviruses. L2 accumulates rapidly in the nucleus after synthesis in the cytoplasm. To identify L2 sequences responsible for nuclear targeting and accumulation, full-length HPV6bL2 protein and L2 proteins containing sequence deletions were expressed in CV-1 cells, using recombinant vaccinia viruses. beta gal protein fused to C terminal and N-terminal L2 sequences were localized in the nucleus and demonstrated that both the C-terminal putative nuclear localization signal (NLS) and N-terminal DNA binding sequences of the L2 protein, which are rich in arginine and lysine, are functional in targeting proteins into the nucleus. L2 mutants lacking amino acids (aa 286-306) do not accumulate in nucleus even when the C-terminal NLS and N-terminal DNA binding sequences are intact. Accumulation in the nucleus of L2 protein lacking aa 286-306 could be also achieved by increasing the L2 protein size via insertion of a portion of L1 protein. Amino acid sequence alignment of L2 proteins from sequenced papillomaviruses showed that sequences in this region are relatively conserved. Our results indicate that in the L2 protein, the nuclear targeting and accumulation are controlled by two different sequences and the rapid nuclear translocation and accumulation may play an important role in papillomavirus life cycle. PMID- 7491756 TI - Immunological adenovirus variant strains of subgenus D: comparison of the hexon and fiber sequences. AB - The fiber and hexon genes of the immunological adenovirus (Ad) variant strains Ad9/Hx and Ad15/Hx, belonging to subgenus D, were sequenced and compared to the corresponding sequences of the prototypes Ad9 and Ad15. It was found that the variants possessed a novel and identical hemagglutinin as they could not be distinguished by hemagglutination-inhibition tests. These serological data were now confirmed on DNA and protein level. The analyzed hexon regions of Ad15 and Ad15/Hx, and Ad9 and Ad9/Hx, respectively, were 100% identical on the amino acid level. The comparison between the variant fibers revealed that they possessed an identical fiber, which was distinct from the Ad9 and Ad15 fiber polypeptides. It seems likely that the donor of the novel fiber protein was a so far unidentified subgroup D adenovirus. The fiber polypeptides of Ad9 and the variants revealed the highest homology (73.7%), whereas the variants and the Ad15 fiber had only 64.2% of the amino acids in common. Most of the differences between the adenovirus serotypes were detected in the fiber knob. Seven conserved sequences found for subgenus D fibers could be confirmed for the fibers of the variants. Furthermore, the data presented complement the knowledge on the organization of subgenus D fiber genes. PMID- 7491759 TI - Sequence of the M28 dsRNA: preprotoxin is processed to an alpha/beta heterodimeric protein toxin. AB - The killer and immunity phenotypes of K28 killer strains of Saccharomyces cerevisiae are determined by the 1.75-kb M28 dsRNA virus. In the plus strand, M28p, the K28 preprotoxin gene, comprises bases 13-1047 and is followed, after an additional 85 bases, by a 63-bp poly(A) sequence and a 553-base 3'-sequence. This 3'-sequence contains two potential stem-loop structures predicted to bind the L-A encoded cap-pol protein, initiating encapsidation; high-level expression results in curing of M1 dsRNA. Expression of M28p confers the complete K28 killer and immunity phenotype on a cell lacking M28 dsRNA. K28 toxin is a disulfide-bonded heterodimer of alpha (10.5 kDa) and beta (11 kDa) components whose N-termini correspond to M28p residues 50-61 and 246-257, respectively. alpha is preceded by a potentially redundant pair of secretion signal peptides; deletion of the first reduces toxin secretion by 75%. While M28p bears no sequence similarity to M1p, the K1 preprotoxin, the predicted patterns of processing by glycosylation and cleavage are remarkably similar. The beta N- and C-termini are probably processed by Kex2p and Kex1p, respectively; the mechanism of cleavage at the less typical sites bounding the alpha component is under investigation. While a kex2 delta mutation prevents toxin secretion, secreted toxin retains 20% activity in a kex1 delta mutant. Neither mutation affects immunity. PMID- 7491760 TI - Mutations in conserved domain I of the Sendai virus L polymerase protein uncouple transcription and replication. AB - To begin to map functional domains of the Sendai P-L RNA polymerase complex we wanted to characterize the P binding site on the Sendai L protein. Analysis of in vitro and in vivo P-L polymerase complex formation with carboxyl-truncations of the L protein showed that the N-terminal half of the protein was required. Site directed mutagenesis of the Sendai virus L gene was employed to change amino acids within a highly conserved region of the N-terminal domain I from amino acids (aa) 348-379 singly or in pairs from the Sendai to the corresponding measles L sequence or to alanine. The mutant L proteins coexpressed with the viral P and NP proteins in mammalian cells were assayed for their ability to form the P-L complex and to synthesize RNA in vitro and showed a variety of defective phenotypes. While most of the mutant L proteins still formed the P-L polymerase complex, a change from serine to arginine at aa 368 and a three-amino-acid insertion at aa 379 virtually abolished both complex formation and RNA synthesis. Changes of aas 370 and 376-377 in the L protein gave only small decreases in viral RNA synthesis. Substitutions at either aas 349-350 or aas 354-355 and a three-amino-acid insertion at aa 348 in the L protein yielded enzymes that catalyzed significant transcription, but were defective in DI RNA replication, thus differentially affecting the two processes. Since DI leader RNA, but not genome RNA, was still synthesized by this class of mutants, the defect in replication appears to be in the ability of the mutant enzyme to package newly synthesized nascent RNA. Single changes at aas 362, 363, and 366 in the L protein gave enzymes with severely decreased overall RNA synthesis, although some leader RNA was synthesized, suggesting that they cannot transcribe or replicate past the leader gene. These studies have identified a region in conserved domain I critical for multiple functions of the Sendai virus L protein. PMID- 7491762 TI - Two strong 5' splice sites and competing, suboptimal 3' splice sites involved in alternative splicing of human immunodeficiency virus type 1 RNA. AB - The human immunodeficiency virus type 1 (HIV-1) genome contains 20 exons that are alternatively spliced from 16 splice sites to generate more than 40 different mRNAs, including incompletely spliced and unspliced mRNAs. In contrast to avian retroviral RNA, which has a cis-acting element in gag that negatively regulates splicing (NRS), HIV-1 RNA did not have any NRS sequences in the gag or pol genes detectable by a splicing inhibition assay. However, this assay demonstrated that the HIV-1 first 5' splice site competed with a cellular 5' splice site, suggesting that HIV-1 may have some strong splice sites. To extend this observation, we used a splice site swapping strategy to determine the efficiency of 14 HIV-1 splice sites in human beta globin chimeras tested in transient transfection experiments. While the 1st HIV-1 5' splice site used in all spliced transcripts and the 4th 5' splice site used in most of the 2-kb transcripts were efficient, the other splice sites, including all the 3' splice sites, were less efficient, ranging in use from 25 to 60%. We propose that this range of splice site efficiencies contributes to the regulation of alternative splicing of HIV-1 mRNAs. PMID- 7491761 TI - Equine arteritis virus subgenomic RNA transcription: UV inactivation and translation inhibition studies. AB - The expression of the genetic information of equine arteritis virus (EAV), an arterivirus, involves the synthesis of six subgenomic (sg) mRNAs. These are 5' and 3' coterminal since they are composed of a leader and a body sequence, which are identical to the 5' and 3' ends of the genome, respectively. Previously, it has been suggested that cis-splicing of a genome-length precursor RNA is involved in their synthesis. This was reevaluated in a comparative analysis of the sg RNA synthesis of EAV, the coronavirus mouse hepatitis virus (MHV), and the alphavirus Sindbis virus. UV transcription mapping showed that the majority of the EAV sg RNAs made at later stages of infection is not derived from a genome-length precursor. However, complete independence of sg RNA synthesis from that of genomic RNA was never observed during the course of infection. The possibility that this resulted from UV irradiation-induced effects on the synthesis of the viral replicase was investigated by inhibiting translation using cycloheximide. For EAV, ongoing protein synthesis was found to be more important for the synthesis of sg RNA than for that of genomic RNA. In general, MHV transcription was extremely sensitive to translation inhibition, whereas EAV genomic RNA synthesis became independent of de novo protein synthesis late in infection. PMID- 7491763 TI - Characterization of a CD4-expressing macaque cell line that can detect virus after a single replication cycle and can be infected by diverse simian immunodeficiency virus isolates. AB - Primate lentiviruses such as human immunodeficiency virus (HIV) and simian immunodeficiency virus (SIV) are phenotypically diverse, and virus isolates vary in cytopathicity, replication rate, and cell tropism. While all virus isolates infect primary peripheral blood lymphocytes, only a subset of strains infect established CD4-expressing T-cell lines. Here, we describe the development and characterization of a macaque cell line that can be infected by all of the strains of SIV that we have tested, including macrophage- and T-cell-tropic strains, primary and cell-line adapted strains, and SIVmac, SIVMne, and SIVsm isolates. The cells can be infected by strains of HIV type 2 (HIV-2) to varying degrees, but not by either cloned or primary isolates of HIV type 1 (HIV-1). This cell line is a derivative of a rhesus macaque mammary tumor cell line (CMMT) engineered to express human CD4. For these studies, a CMMT-CD4 clone expressing an integrated copy of a truncated HIV-1 long terminal repeat fused to the beta galactosidase gene (LTR-beta-gal) was established to allow detection of infectious SIV after a single round of replication. Here, we demonstrate the ability of the CMMT-CD4-LTR-beta-gal cell line to rapidly and quantitatively detect infectious SIV. Using these cells to assay virus, we could readily measure neutralizing antibody activity in animals infected with different SIV isolates. Neutralizing activity was detected against the homologous virus and lower, but detectable, activity was measured against heterologous virus. Thus, this system, which is highly sensitive and can detect infection by all of the SIV isolates we tested, is a rapid method for detecting infectious virus and quantitating neutralizing antibody activity. PMID- 7491764 TI - HERV-H endogenous retroviruses: presence in the New World branch but amplification in the Old World primate lineage. AB - The evolutionary origin and age of the HERV-H family of human endogenous retrovirus-like sequences was investigated in this study. HERV-H elements exist in approximately 900 partially deleted copies and 50-100 more intact forms in humans and Old World monkeys. However, their possible presence in more divergent species is unknown. We have isolated a 1.6-kb genomic DNA segment from the New World monkey marmoset that had been PCR amplified using human HERV-H primers. DNA and protein comparisons and database searches indicate that this marmoset clone is more closely related to human HERV-H elements than to any other sequence, indicating that HERV-H-related sequences do exist in New World monkeys. In contrast to the high copy numbers of deleted elements in Old World primates. Southern blot analysis shows that such elements are present in less than 50 copies in two different species of New World monkey. To estimate evolutionary ages of the common deleted form of the element, a selected DNA segment from the pol region was compared from multiple human HERV-H elements. This comparison suggests that many HERV-H elements of the abundant deleted subfamily integrated approximately 30-35 million years ago. Very similar percentage divergence values between 5' and 3' long terminal repeats of individual elements of the deleted subfamily also suggest that these elements are close in age. These results indicate that HERV-H elements first appeared in the germline prior to the New World/Old World divergence over 40 million years ago. Interestingly, they remained in low numbers in the New World branch while a subfamily underwent a major amplification in Old World primates before the time of divergence of hominoids from Old World monkeys. PMID- 7491765 TI - Phosphorylation of Ser232 directly regulates the transcriptional activity of the P protein of human respiratory syncytial virus: phosphorylation of Ser237 may play an accessory role. AB - The phosphoprotein P of human respiratory syncytial virus (RSV) was expressed in eukaryotic cells in phosphorylated form. Site-directed mutagenesis of the recombinant protein established Ser232 as the major site of phosphorylation in vivo. Phosphorylation of bacterially made P protein in vitro by purified casein kinase II (CKII) resulted in the phosphorylation of Ser237, whereas mainly Ser232 was phosphorylated by a crude cell extract. The P kinase activity in the cell extract exhibited properties characteristic of CKII. While the Ser232,237 to Ala double mutant was nearly completely defective for phosphorylation and transcription, phosphorylation at Ser232, through the use of appropriate P mutant or kinase, activated P protein. Phosphorylation of Ser237 restored activity only to the extent it facilitated phosphorylation of Ser232. Phosphate groups of P protein in RSV-infected cells were highly stable; inhibitors of protein serine phosphatases had no effect on the intracellular turnover of the phosphates. Highly purified viral polymerase L was transcriptionally active but devoid of P protein kinase activity. Thus, CKII-mediated phosphorylation of Ser232 appears to be the primary regulator of P protein activity while phosphorylation of Ser237 may be involved in a modulatory role under certain conditions. PMID- 7491766 TI - HIV-1 Tat directly interacts with the interferon-induced, double-stranded RNA dependent kinase, PKR. AB - We present evidence that the HIV-1 Tat protein and the RNA-dependent cellular protein kinase, PKR, interact with each other both in vitro and in vivo. Using GST fusion chromatography, we demonstrate that PKR, interacts directly with the HIV-1 Tat protein. The region in Tat sufficient for binding PKR maps within amino acids 20 to 72. In in vitro assays, the two-exon form of Tat (Tat 86) was phosphorylated by PKR, while the one exon form of Tat (Tat 72) inhibited PKR autophosphorylation and substrate phosphorylation. The ability of Tat to interact with PKR was demonstrated in both yeast and mammalian cells. Expression of PKR in yeast results in a growth suppressor phenotype which was reversed by coexpression of a one exon form of Tat. Expression of Tat 72 in HeLa cells resulted in direct interaction with PKR as detected by coimmunprecipitation with a Tat antibody. Tat and PKR also form a coimmunoprecipitable complex in cell-free extracts prepared from productively infected T lymphocytes. The interaction of Tat with PKR provides a potential mechanism by which HIV could suppress the interferon system. PMID- 7491767 TI - Tomato bushy stunt virus spread is regulated by two nested genes that function in cell-to-cell movement and host-dependent systemic invasion. AB - We have investigated the importance of two small nested genes (p19 and p22) located near the 3' end of the genome of tomato bushy stunt virus (TBSV) for infectivity in several hosts. Our results show that both genes are dispensable for replication and transcription and that the p19 gene encodes a soluble protein, whereas the p22 gene specifies a membrane-associated protein. Assays using TBSV derivatives that have the beta-glucuronidase gene substituted for the capsid protein gene demonstrate that p22 is required for cell-to-cell movement in all plants tested. Mutations inactivating p19 ameliorate the severe necrotic systemic symptoms elicited by wild-type TBSV in Nicotiana benthamiana and Nicotiana clevelandii, but p19 does not obviously affect movement in these hosts. However, in some local lesion hosts p19 influences the lesion diameter, which suggests that it has an auxiliary host-dependent role in movement. This notion is supported by the observation that p19 is required for long-distance spread of TBSV in spinach and for systemic infection of pepper plants. Thus, movement of TBSV is regulated by two nested genes; p22 governs cell-to-cell movement and p19 has a host-specific role in systemic invasion. PMID- 7491769 TI - HIV-1 mediates rapid apoptosis of lymphocytes from human CD4 transgenic but not normal rabbits. AB - Normal rabbit lymphocytes can be infected with HIV-1 although infection is much less efficient than in human lymphocytes. When peripheral blood mononuclear cells (PBMC) from rabbits transgenic for human CD4 (HuCD4) were exposed to HIV-1, enhanced infection and a rapid depletion of lymphocytes were observed. Cell death in the infected cultures occurred via apoptosis, but no similar effect was seen in nontransgenic rabbit PBMC cultures. Induction of apoptosis in HuCD4-expressing cells required virus replication; heat-inactivated virus or recombinant viral proteins had no effect on cell viability. Expression of the Fas antigen was increased in HIV-1-infected CD4+ rabbit lymphocytes. Characterization of the infected PBMC cultures revealed that apoptosis occurs both in HuCD4+ and HuCD4- cells, indicating that bystander cells are killed. These data define a requirement for HuCD4 in initiation, but not the spread, of HIV-1-induced apoptosis in rabbit PBMC and provide a model to probe mechanisms leading to lymphocyte depletion in HIV-1 infection. PMID- 7491768 TI - Analysis of trafficking of Rev and transdominant Rev proteins in living cells using green fluorescent protein fusions: transdominant Rev blocks the export of Rev from the nucleus to the cytoplasm. AB - Expression of gag/pol and env genes of human immunodeficiency virus requires the viral Rev protein. Mutant Rev proteins, displaying a transdominant phenotype (TDRev), were shown to inhibit Rev function. To investigate the underlying mechanism of this inhibition, the green fluorescent protein (GFP) of Aequorea victoria was fused to Rev and TDRev, which allowed the study of their trafficking and interactions in living human cells. Both Rev-GFP and TDRev-GFP were shown to retain appropriate nucleolar localization and function. Upon actinomycin D treatment, Rev-GFP was transported to the cytoplasm within 1.5 hr, while TDRev, although partially dissociated from the nucleolus, was retained in the nucleus. Coexpression of Rev-GFP and TDRev in the same cell demonstrated that TDRev inhibited the transport of Rev-GFP from the nucleus to the cytoplasm. This inhibition was specific for Rev, since the export of the functionally analogous Rex protein of the human T-cell leukemia virus type I was not inhibited by TDRev. These results indicate that Rev and TDRev form heteromultimers in the nucleolus and that this interaction prevents Rev's export from the nucleus to the cytoplasm. In addition to providing a model for the function of TDRev, these results also demonstrate the successful application of protein fusions to GFP to study localization and trafficking of proteins in living mammalian cells. PMID- 7491770 TI - Differential subcellular localization of hepatitis C virus core gene products. AB - The expression of the core gene of two different hepatitis C virus (HCV) isolates was analyzed. In the presence of its downstream E1 envelope protein sequence, two major core protein products with molecular masses of 21 kDa (P21) and 19 kDa (P19) and a minor protein product with molecular mass of 16 kDa (P16) were detected. In the absence of its downstream E1 envelope protein sequence, P21 and P19 remained the major protein products expressed from the core gene of the HCV RH isolate, whereas P16 became the major protein product of the core gene of the HCV-1 isolate. Analysis of the amino-terminal sequences of P21 and P16 expressed in Escherichia coli revealed that P21 and P16 were co-amino terminal. Deletion mapping analysis indicated that P16 lacked the carboxy-terminal sequence of P21. Immunofluorescence analysis of the subcellular localization of different HCV core proteins indicated that P21 and P19 displayed a reticular and punctate staining pattern typical of endoplasmic reticulum-associated proteins, while P16 was localized to the nucleus. The distinct subcellular localization of P16 raises the possibility that P16 may have a biological function very different from those of P21 and P19. PMID- 7491771 TI - Kinetic and functional mapping of viral epitopes using biosensor technology. AB - Some monoclonal antibodies (Mabs) that react with the extremity of the tobacco mosaic virus (TMV) particle containing the 5' end of the RNA are able to block the disassembly of TMV by ribosomes while others are totally devoid of such activity. No correlation could be established between the binding kinetics and affinity of the Mabs and their inhibitory capacity. An epitope map of the Mab binding sites was constructed on the basis of kinetic two-site binding assays with the viral monomeric protein (TMVP) performed using biosensor technology (BlAcore). Mabs possessing inhibitory activity were found to bind to the part of the TMVP surface closest to the central axis in the polymerized particle. As this part of the subunit is known to interact with the viral RNA, it seems that inhibitory Mabs act by sterically preventing the interaction between virus and ribosomes. This study illustrates the advantages of the biosensor technology for locating conformational epitopes in viral proteins. PMID- 7491772 TI - Expression of tobacco ringspot virus capsid protein and satellite RNA in insect cells and three-dimensional structure of tobacco ringspot virus-like particles. AB - The capsid protein gene of tobacco ringspot virus (TobRV), which had been modified to contain an amino-terminal methionine codon, was ligated into a baculovirus transfer vector downstream from the polyhedrin promoter. The resulting plasmid was cotransfected with linearized baculovirus DNA into insect cells. Recombinant baculovirus expressed high levels of the TobRV capsid protein that assembled to form virus-like particles that were similar in size and shape to authentic TobRV capsids. These virus-like particles did not encapsidate any RNA, including the capsid protein mRNA. The capsid protein mRNA is a truncated RNA 2, which may lack a putative encapsidation signal. To determine whether an intact packaging substrate could be encapsidated by the TobRV capsid protein, another recombinant baculovirus, concomitantly expressing both capsid protein and TobRV satellite RNA, was constructed. Surprisingly, the vast majority of the satellite RNA molecules expressed from this recombinant baculovirus were ligated in the insect cells to form circular RNA molecules. Like circular forms of satellite RNA generated in planta, these circular satellite molecules remained unencapsidated by the TobRV capsid protein. Computer-generated three-dimensional reconstruction using electron cryomicrographs of the empty virus-like particles allowed the first structural analyses of any nepovirus capsid. This 22-A resolution reconstruction resembled capsids of other members of the picornavirus superfamily. These data support the hypothesis that the nepovirus capsid is structurally analogous to those of the como- and picornaviruses. PMID- 7491773 TI - Activation of astrocytes in the spinal cord of mice chronically infected with a neurotropic coronavirus. AB - Mice infected with the neurotropic JHM strain of mouse hepatitis virus (MHV-JHM) develop a demyelinating encephalomyelitis several weeks after infection. Astrogliosis and infiltration of inflammatory cells are prominent findings in the brains and spinal cords of infected mice. In this report, astrocytes in infected spinal cords were analyzed for expression of three pleiotropic cytokines, TNF alpha, IL-1 beta, and IL-6; Type 2 nitric oxide synthase (iNOS); and MHC class I and II antigen. The data show that all three cytokines and iNOS are expressed by astrocytes in chronically infected spinal cords. These activated astrocytes are localized to areas of virus infection and demyelination, although most of the astrocytes expressing these proteins are not MHV-infected. MHC class I and II antigen can be detected in these spinal cords as well, but not in cells with the typical morphology of astrocytes. TNF-alpha, IL-6, and iNOS are also evident in the brains of mice with MHV-induced acute encephalitis, but in marked contrast to the results obtained with the chronically infected mice, most of the cells expressing these cytokines or iNOS had the morphology of macrophages or other mononuclear cells and very few appeared to be astrocytes. Additionally, astrocytes and, most likely, oligodendrocytes are infected in the spinal cords of mice with chronic demyelination. These results are consistent with a role for both viral infection of glial cells and high localized levels of proinflammatory cytokines and nitric oxide in the demyelinating process in mice infected with MHV JHM. They also show that analogously to the human demyelinating disease, multiple sclerosis, astrocytes are a major cellular source for these cytokines in mice with chronic, but not acute disease. PMID- 7491774 TI - The adenovirus protease is required for virus entry into host cells. AB - We have analyzed the mechanisms used by adenovirus to gain entry into the host cell. Using both virus infection and the ability of adenovirus particles to enhance polylysine/DNA uptake as a measure of virus entry, we have demonstrated that the adenovirus-encoded 23K protease is required for two functions in the infection process. A proteolytic processing of the capsid is required to generate a virus capsid that can increase membrane interactions at pH 5. We have found that forms of adenovirus capsid that have not undergone the processing reactions (immature capsids) are deficient in their ability to disrupt membranes at pH 5 and are unable to enhance the entry of polylysine/DNA complexes. A second role of the protease was revealed by experiments using inhibitors of the protease. Mature virus capsids lose their ability to enhance gene delivery and become noninfectious after exposure to inhibitors of the protease (1 microM N ethylmaleimide, 100-300 microM copper chloride, 1 microM MDL28170, or anti protease antiserum), suggesting that the viral protease activity is required during the cellular entry process. PMID- 7491775 TI - Induction of antibodies protecting against transmissible gastroenteritis coronavirus (TGEV) by recombinant adenovirus expressing TGEV spike protein. AB - Ten recombinant adenoviruses expressing either fragments of 1135, 1587, or 3329 nt or the full-length spike gene of transmissible gastroenteritis coronavirus (TGEV) have been constructed. These recombinants produce S polypeptides with apparent molecular masses of 68, 86, 135, and 200 kDa, respectively. Expression of the recombinant antigen driven by Ad5 promoters was inhibited by the insertion of an exogenous SV-40 promoter. Most of the recombinant antigens remain intracytoplasmic in infected cells. All the recombinant-directed expression products contain functional antigenic sites C and B (Gebauer et al., 1991, Virology 183, 225-238). The recombinant antigen of 135 kDa and that of 200 kDa, which represents the whole spike protein, also contain antigenic sites D and A, which have previously been shown to be the major inducers of TGEV-neutralizing antibodies. Interestingly, here we show that recombinant S protein fragments expressing only sites C and B also induced TGEV-neutralizing antibodies. The chimeric Ad5-TGEV recombinants elicited lactogenic immunity in hamsters, including the production of TGEV-neutralizing antibodies. The antisera induced in swine by the Ad5 recombinants expressing the amino-terminal 26% of the spike protein (containing sites C and B) or the full-length spike protein, when mixed with a lethal dose of virus prior to administration to susceptible piglets, delayed or completely prevented the induction of symptoms of disease, respectively. PMID- 7491776 TI - Expression, purification, and identification of a novel self-cleavage site of the Nla C-terminal 27-kDa protease of turnip mosaic potyvirus C5. AB - The gene encoding the C-terminal protease domain (27 kDa) of the nuclear inclusion protein a of turnip mosaic potyvirus C5 was cloned and expressed as a fusion protein with glutathione S-transferase in Escherichia coli XL1-blue. Two forms of the protease (27 and 25 kDa) were purified from the fusion protein by glutathione affinity chromatography and Mono S chromatography and exhibited the specific proteolytic activity when a synthetic undecapeptide, Glu-Pro-Thr-Val-Tyr His-Gln-Thr-Leu-Asn-Glu, or an in vitro translation product of the polyprotein containing the cleavage site between the nuclear inclusion protein b and the capsid protein, was used as a substrate. The purified proteases showed a Km of 1.15 +/- 0.16 mM and a Vmax of 0.74 +/- 0.091 mumol/mg/min with the synthetic peptide substrate. The 25-kDa protein was found to be generated by the cleavage between Ser223 and Gly224 near the C-terminus of the 27-kDa protease and to retain the specific proteolytic activity. The point mutation of Asp81 or Cys151, two putative active site residues in the 27-kDa protease, to Asn or Ser, respectively, prevented the generation of the 25-kDa protein and diminished the proteolytic activity of the protease drastically, suggesting that the 27-kDa protease cleaves itself between Ser223 and Gly224 to generate the 25-kDa protein. PMID- 7491777 TI - Transformed mouse cell lines that consist predominantly of cells maintaining bovine papilloma virus at high copy number. AB - Rare cells that contain large amounts of bovine papilloma virus (BPV) DNA have been observed in populations of BPV-transformed mouse ID13 cells. The viral DNA molecules in these "jackpot cells" have been thought to have switched from the controlled replication typical of latent BPV infection to the uncontrolled "runaway" prelytic replication characteristic of terminal stage infection of bovine epidermal cells. By sequential subcloning of high-BPV derivatives of ID13, we isolated stable cell lines enriched more than 1000-fold for cells showing large amounts of BPV by fluorescence in situ hybridization analysis. High-BPV subclones contained a variant plasmid as well as wild-type BPV DNA; analysis of the BPV variants in two independently isolated subclones that showed the high-BPV phenotype in 50 to 80% of cells in the population indicated that both variants had undergone tandem duplication of the BPV long control region, which contains the viral origin of replication and transcription enhancer sequences. Transfer of the high-copy-number phenotype by transfection of DNA from high-BPV cells was accompanied by transfer of the variant plasmid. Density gradient analysis of BPV plasmid replication in high-BPV subclones showed the random-choice mode of replication observed in the parental ID13 population, rather than the rapid BPV replication found in epidermal cells destined for lysis and death. Our results indicate that high-BPV cells in actively dividing ID13 populations are not produced by uncontrolled replication of viral DNA and suggest that they may result instead from abnormal plasmid partitioning. PMID- 7491778 TI - Limited viral spread and rapid immune response in lymph nodes of macaques inoculated with attenuated simian immunodeficiency virus. AB - A comparative study was undertaken to characterize the very early events that distinguish attenuated and pathogenic simian immunodeficiency virus (SIV) infections. Three rhesus macaques were inoculated with the attenuated SIVmac 251 delta nef virus, and three others with a virus of intermediate phenotype, SIVmac 239 nef stop. They were compared to four macaques inoculated with the pathogenic SIVmac 251 isolate. Lymph nodes (LN) taken between 7 days and 2 months postinoculation were analyzed for SIV expression by in situ hybridization. During acute infection, SIV 21 delta nef infected 1 to 1.5 log10 fewer cells in LN tissue than the pathogenic SIV 251 isolate. The reduction was more marked in the blood, as SIV 251 delta nef infected 2 to 3 log10 fewer PBMC than the isolate and did not yield detectable antigenemia. Morphometric measurements showed that the development of germinal centers (GC) was more rapid in the delta nef infection, which led to a more efficient trapping of viral particles, and could account for antigenemia clearance. The SIV 239 nef stop clone reverted to a nef+ genotype at Week 2, but induced a lower viral burden than a directly pathogenic virus. The kinetics of GC development was rapid, indicating that SIV 239 nef stop induced an immune response similar to that seen in attenuated infection. This study provides evidence that attenuated SIV elicits a more rapid immune response than pathogenic SIV and suggests that an early immunosuppressive episode may facilitate the dissemination of pathogenic SIV. PMID- 7491779 TI - Picornavirus 2A proteinase-mediated stimulation of internal initiation of translation is dependent on enzymatic activity and the cleavage products of cellular proteins. AB - Poliovirus and human rhinovirus 2A proteinases are known to stimulate translation initiation on the cognate viral Internal Ribosome Entry Segments (IRESes). The molecular mechanism of this translational transactivation was investigated in vitro using dicistronic mRNAs containing picornaviral IRESes as the intercistronic spacer and purified human rhinovirus type 2 and coxsackievirus B4 2A proteinases. The stimulation achieved on the HRV2 IRES in the presence of the cognate 2A proteinase at 1 microgram/ml was twofold; the maximum stimulation at 100 micrograms/ml was fivefold. The IRESes and proteinases from rhino- and enteroviruses were interchangeable; however, stimulation of translation initiation on a cardiovirus IRES by these proteinases was minimal. Studies using an inhibitor or a mutant 2A proteinase demonstrated that translation stimulation requires 2A-mediated enzymatic conversion of some cellular component(s). The HRV2 2A proteinase also stimulated translation initiation on full-length viral RNA, suggesting that 2A proteinase-mediated stimulation of IRES-driven translation has a physiological role. PMID- 7491780 TI - Molecular basis of antigenic variation between the glycoproteins C of respiratory bovine herpesvirus 1 (BHV-1) and neurovirulent BHV-5. AB - Herpesvirus glycoprotein C (gC) functions as a major virus attachment protein. The gC sequence of the neurovirulent bovine herpesvirus type 5 (BHV-5) virus was determined and compared with the gC sequence of the nonneurovirulent BHV-1. Alignment of the predicted amino acid sequences of BHV-1 and BHV-5 gC ORFs showed that the amino-terminal third of the protein differed between the two viruses. Whole or subgenomic fragments of gC coding regions from both viruses were expressed as trpE-gC fusion proteins in Escherichia coli to map linear epitopes defined by type-specific murine monoclonal antibodies (MAbs). Based on the reactivity of BHV-1-specific MAbs with the recombinant proteins, two epitopes were mapped between BHV-1 gC residues 22 and 172. Undirectional deletion of these residues at the carboxy end mapped one within residues 22-69 and the other within residues 103-122. Two BHV-5-specific MAbs identified an epitope coding region within BHV-5 gC residues 31-78. Bovine antisera against BHV-1 and BHV-5 showed specificity to BHV-1 gC residues 22-69 and to BHV-5 gC residues 31-78, respectively, in a type-specific manner. PMID- 7491781 TI - Experimental evidence of recombination in coronavirus infectious bronchitis virus. AB - Embryonated eggs were coinfected with two strains of the coronavirus avian infectious bronchitis virus (IBV), IBV-Beaudette and IBV-M41, to investigate whether recombination between the two strains would occur. Virions were isolated from the allantoic fluid of the coinfected eggs and putative hybrid RNAs were detected by polymerase chain reaction (PCR), using strain-specific oligonucleotides. PCR products, of the expected sizes, were obtained as predicted from potential recombination events between the nucleoprotein (N) gene and the 3' untranslated region of the two IBV genomes. Sequencing confirmed that they corresponded to hybrid RNAs. Virus produced as a result of the mixed infection was treated with an M41-specific neutralizing monoclonal antibody and passaged in Vero cells, in which IBV-Beaudette, but not IBV-M41, replicated. Hybrid RNA was still detectable after three serial passages. Since no IBV-M41 was detectable this confirmed that infectious recombinant genomes had been produced in the embryonated eggs. These findings not only support the circumstantial evidence, from sequencing studies of IBV field strains, that recombination occurs during replication of IBV and contributes to the diversity of IBV, but also show that coronavirus RNA recombination is not limited to mouse hepatitis virus. PMID- 7491782 TI - Identification of the active-site residues of the 3C proteinase of foot-and-mouth disease virus. AB - To identify the active-site residues of the 3C proteinase of foot-and-mouth disease virus (FMDV), we introduced mutations into the 3C coding region and examined the activity of mutant enzymes on various substrates. Based on alignment of FMDV 3C with other picornavirus 3C proteinases and with the trypsin family of serine proteinases, mutations were introduced at residues presumed to be part of the catalytic triad, involved in substrate binding, or present in nonconserved regions. Wild-type and mutant 3C proteins were expressed in Escherichia coli and tested for their ability to cleave synthetic substrates corresponding to different portions of the viral genome. Substitutions at His-46 (catalytic triad), Asp-84 (catalytic triad), or His-181 (substrate binding) produced enzymes unable to process P1, P2, or P3 substrates in trans, whereas a change in the conserved Asp-98 had no effect on enzyme activity. Substitution of Ser for Cys 163 (catalytic triad) yielded an enzyme that retained activity on some substrates, while a substitution of Gly at this position resulted in a completely inactive enzyme. The kinetics of trans processing of translation products from a transcript encoding the P1 and P2 coding regions and the 2C/3A cleavage site with wild-type 3C or a transcript encoding P1 with 3C mutants revealed that the order of cleavage was VP3-VP1, VP0-VP3, VP1-2A, 2C-3A, and 2B-2C. Mutations in 3C that resulted in a partially active enzyme were individually introduced into full length FMDV cDNA and RNA transcripts were translated in a cell-free system and used to transfect cells. In all cases the virus that was rescued had reverted to the wild-type 3C codon. PMID- 7491783 TI - Isolation and characterization of Marek's disease virus (MDV) cDNAs mapping to the BamHI-I2, BamHI-Q2, and BamHI-L fragments of the MDV genome from lymphoblastoid cells transformed and persistently infected with MDV. AB - We have isolated and sequenced two cDNAs of sizes 2674 and 677 bp from a cDNA library derived from MKT-1, a lymphoblastoid cell line transformed and latently infected with Marek's disease virus (MDV) using probes corresponding to the right hand end of the BamHI-I2 fragment of the MDV genome. The larger cDNA clone represents an abundant transcript, which extends from the right-hand end of BamHI I2 to the adjacent BamHI-Q2 and BamHI-L fragments of the MDV genome and contains the Meq (MDV Eco-Q) open reading frame. The smaller cDNA clone represents a spliced transcript containing the putative DNA binding domain of Meq as well as sequences in the BamHI-L region. We prepared a polyclonal antibody against part of the protein sequence of Meq and detected a 44-kDa protein in MKT-1 cells and in cells lytically infected with MDV. In addition, riboprobes corresponding to sequences specific to each cDNA as well as shared sequences between cDNAs detected a number of transcripts in cells either lytically or latently infected with MDV. Our results indicate that the Meq transcriptional unit extends to the BamHI-L fragment and that the transcripts mapping to the right-hand end of the BamHI-I2 and adjacent BamHI-Q2 and BamHI-L fragments are not preferentially expressed during latency. PMID- 7491784 TI - Lymphocyte-tropic simian immunodeficiency virus causes persistent infection in the brains of rhesus monkeys. AB - Molecularly cloned SIVmac239 is the prototypical SIVmac lymphocyte-tropic virus that replicates productively in lymphocytes but poorly in macrophages. In macaques, the virus causes activation and productive infection of T lymphocytes which invade the central nervous system (CNS) early after infection in the animal. However, infected animals develop immunosuppression and AIDS but rarely overt neurological disease. In this study, we examined multiple regions of the brain and spinal cord for the presence of SIV env sequences and histological lesions in five macaques that had been infected with SIVmac239 for 1.7 to 2.25 years. Histopathological examination of the brain revealed no lesions consistent with encephalitis; however, viral DNA was found in all five brains. In one animal the virus caused infection in a widely disseminated pattern from the frontal cortex to the distal end of the spinal cord, whereas in the other four animals infection in the CNS occurred in a nonspecific, focal pattern. Sequence analyses were performed on gp120 sequences isolated from selected regions of the CNS and compared to gp120 sequences isolated from corresponding lymph nodes, a tissue known to support productive replication of SIVmac239. Examination of the viral sequences from the CNS tissue from two animals (macaques 10F and 14F) revealed a low mutation rate when compared to the sequences isolated from the lymph node tissues. The percentage change in the amino acid sequence was approximately 1% for CNS clones versus > or = 3% for clones isolated from the lymph node. The majority of the CNS viral sequences of macaques 10F and 14F had none of the genetic markers shown in a previous study to be associated with macrophage-tropic variants and indeed retained a nucleotide sequence of similar to the original lymphocyte-tropic virus used for inoculation despite almost 2 years of persistent infection in the animals. Construction of chimeric viruses with V1-V5 regions of selected macaque 10F and macaque 14F CNS-gp120 clones confirmed the predicted lymphocyte-tropic nature of these env genes. In contrast, the gp120 sequences isolated from the CNS tissue of one of the other three animals (macaque 13F) had a mutation rate comparable to that observed for the lymph node clones. The CNS clones from this animal had amino acid substitutions that were previously shown to be associated with macrophage tropism. Compared to the chimeric viruses constructed with V1-V5 sequences from macaques 10F and 14F, viruses constructed with the V1-V5 sequences of several macaque 13F brain clones did not yield infectious virus.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 7491785 TI - Intracellular localisation of herpes simplex virus type 1 ribonucleotide reductase subunits during infection of cultured cells. AB - We have analysed the intracellular localisation of herpes simplex virus type 1 ribonucleotide reductase during infection of cultured cells by indirect immunofluorescence using polyclonal and monoclonal antibodies specific for the R1 and R2 subunits. Three different viruses were used to infect cells, wild-type strain 17+ and two temperature-sensitive mutants, ts 1222, which produces R1 only, and ts 1207, which expresses a normal R2 and an altered R1 that fails to interact with R2 at the nonpermissive temperature because of an amino acid substitution in R1. R1 was detected 2 hr postinfection with all three viruses and remained evenly distributed throughout the cytoplasm. R2 was not observed until 4 hr postinfection and, in contrast to the even distribution of R1, was localised in discrete cytoplasmic foci close to the nucleus. In double-labelling experiments both R1 and R2 were found in these foci where they presumably associate to form the active enzyme. As expected R2 was not detectable in cells infected with ts 1222. In ts 1207-infected cells it formed wild-type-like foci, indicating that interaction with R1 is not required for R2 focus formation. R1 was present in a twofold excess over R2 in wild-type-infected cells. We suggest that the uncomplexed R1 could perform a role associated with the protein kinase present in the N-terminal domain. PMID- 7491786 TI - Induction by herpes simplex virus of free and heteromeric forms of E2F transcription factor. AB - We have determined that HSV causes rapid and large increases in cell-cycle regulated free E2F and S-phase p107/E2F DNA binding activities in asynchronous cultures of C33A cells. Induction occurred by 4 hr postinfection and coincided with the appearance of viral encoded immediate-early and delayed-early proteins, i.e., when viral DNA replication normally commences. No increase in E2F activities occurred when cells were infected with viruses expressing mutant regulatory proteins ICP4 or ICP27, or mutant replication proteins ICP8, pol or helicase, or when cells were infected with wild-type virus in the presence of inhibitors of DNA synthesis. In contrast, ICP8 mutant-infected cells contained elevated amounts of NF kappa B activity equivalent to WT virus, no induction of Sp1 relative to WT virus, and reduced ATF/CREB activity relative to WT virus. Results of transient expression assays with E2F-responsive reporters indicated that the net effect of induction of both active (free E2F) and repressive (p107/E2F) complexes was a decrease in AdE2 promoter activity and an increase in c-myc promoter activity. Taken together these results suggest that HSV can cause unscheduled changes in the amount and functional status of a cell-cycle-regulated transcription factor. These results are discussed in light of possible roles for viral-induced alterations in E2F, especially as related to imposing or overriding cell-cycle checkpoints. PMID- 7491787 TI - The human immunodeficiency virus type 1 Vpu protein: a potential regulator of proteolysis and protein transport in the mammalian secretory pathway. AB - HIV-1 Vpu is a small transmembrane phosphoprotein of 16 kDa which performs critical roles in CD4 proteolysis and virus release. Previous studies have demonstrated that Vpu-induced degradation of CD4 occurs in the endoplasmic reticulum (ER), and that the proteolytic process is sequence specific requiring both the transmembrane and cytoplasmic domains of CD4. In the present study, we investigated the effects of Vpu expression on the intracellular membrane trafficking pathway of mammalian cells. In singly transfected cells, the HIV envelope glycoproteins and vesicular stomatitis virus glycoprotein (VSV G) were properly transported to the cell surface undergoing oligosaccharide modifications characteristic of their movement through the Golgi complex. In contrast, the cell surface delivery of glycoproteins was severely impeded in cells expressing Vpu. Biochemical analyses revealed that Vpu expression blocked the transfer of proteins from the ER-Golgi complex to the plasma membrane in a dose- and protein dependent manner. Soluble gp120 exhibited extreme transport defects in the presence of Vpu, whereas transmembrane proteins (e.g., gp160, VSV) responded only moderately to wild-type Vpu. To gain insight into Vpu-mediated transport inhibition, we performed mutational analysis of the CK-2 phosphorylation sites (serines at 52 and 56) in the Vpu protein. CK-2 phosphorylation of Vpu has been shown to regulate the activity of the protein in reactions that involve the proteolysis of CD4 in the ER. We demonstrate here that the phosphorylation mutant is defective in both sequence-specific degradation of VRE-containing substrates and the transport inhibition of gp120 and VSV-G in the secretory pathway. Thus, these experiments have revealed that Vpu-mediated proteolysis and transport inhibition are mechanistically coupled requiring the same structural elements of the Vpu protein in both processes. We propose that the primary effect of Vpu expression is to impede the secretion process and then access glycoproteins bearing the VRE for Vpu-mediated proteolysis in the ER of mammalian cells. PMID- 7491789 TI - Replication of molluscum contagiosum virus. AB - Molluscum contagiosum virus (MCV) infects preadolescent children and sexually active adults, frequently causing a disfiguring cutaneous disease in immunosuppressed HIV-infected individuals. The development of an efficacious treatment regime has been hampered by the failure to replicate the virus in the laboratory. Here we report the first demonstration of MCV replication in an experimental system. In human foreskin grafts to athymic mice, MCV induced morphological changes which were indistinguishable from patient biopsies and included the development and migration of molluscum bodies containing mature virions to the epidermal surface. PMID- 7491788 TI - Interaction between RNA-dependent RNA polymerase of alfalfa mosaic virus and its template: oxidation of vicinal hydroxyl groups blocks in vitro RNA synthesis. AB - In the life cycle of a (+)-strand RNA plant virus the processes of template RNA recognition and initiation of the synthesis of a complementary strand by the viral RNA-dependent RNA polymerase (RdRp) are crucial early steps. Using a template-dependent in vitro RNA synthesizing system of alfalfa mosaic virus (AIMV) we were able to study the effect of small chemical modifications of the 3' end of the template RNAs on product formation. After oxidation of the 3'-terminal nucleoside of the template no products could be detected. Presumably, RNA synthesis was blocked at the stage of initiation, since the promoter of the RdRp is internal (A. C. Van der Kuyl et al., Virology 176, 346-354, 1990). Blocking was probably due to an irreversible binding of the enzyme to the 3' end of the modified RNA. Using this system it was shown that in template competition experiments the RdRp of AIMV displays a high specificity for its cognate template, either before or after the oxidation of the 3'-terminal nucleoside. From this it was concluded that periodate modification of the 3'-terminal nucleoside has little or no effect on template recognition. Furthermore, we showed that the viral coat protein, which forms a part of the viral polymerase (R. Quadt et al., Virology 182, 309-315, 1991), was not the main target involved in the inhibition of RNA synthesis. PMID- 7491790 TI - Evidence that N-linked glycosylation is necessary for hepatitis B virus secretion. AB - Human hepatitis B virus (HBV) envelopes contain three distinct glycoproteins called L, M, and S HBsAg. Each is posttranslationally modified to contain N linked oligosaccharides. N-linked oligosaccharides, after attachment to a polypeptide backbone, are processed by enzymes within the endoplasmic reticulum (ER). There is uncertainty about what role, if any, these N glycans and their modification in the ER play in the function of the HBV envelope proteins. By treating hepatoblastoma cultures which secrete HBV (HepG 2.2.15 cells) with inhibitors of different steps of the glycosylation and glycan modifying pathway, we provide evidence that glycosylation and the first step in the processing pathway are necessary for virion, but not subviral particle, secretion. That is, using a highly sensitive immunoprecipitation/polymerase chain reaction system, enveloped HBV could not be detected in the medium of HepG2.2.15 cells incubated with tunicamycin. However, HBV subviral particle secretion was not prevented by tunicamycin. Moreover, inhibitors of alpha-glucosidase I (the first step in the glycan processing pathway) also prevented virion secretion. Inhibitors of mannose trimming (a later step) and glycolipid synthesis, did not prevent virion secretion, defining the limits of the glycosylation requirements in secretion. These results demonstrate a requirement for N-glycosylation and glucosidase processing in the secretion of virions and further distinguish between the requirements for virion and subviral particle secretion. PMID- 7491791 TI - Phenotypic determinants of adenovirus E1A gene autoregulation: variable region between conserved coding domains 2 and 3. AB - Different serotypes and evolutionary variants of human adenoviruses exhibit distinctive patterns of positive and negative autoregulation of the viral E1A gene. An autoregulatory E1A promoter mutation of the adenovirus type 3 (Ad3) E1A gene renders Ad3hr15 incapable of growth in normally permissive cells. The promoter mutation is complemented in trans by E1A products of the heterologous helper adenovirus type 5 (Ad5). Second-site revertants of Ad3hr15 restore viability with high levels of E1A gene expression. The revertant E1A genotypes retain the mutant E1A promoter and have small in-frame deletions in the nonconserved region between the repression- and activation-associated conserved domains CR2 and CR3. Plasmid expression vectors were constructed as 12S and 13S cDNA forms of revertant E1A genes. These were used in cotransfection experiments with a reporter gene plasmid under transcriptional control of the mutant Ad3hr15 E1A promoter. The repression of the Ad3hr15 E1A promoter by helper Ad5 or revertant 12S E1A cDNA was consistently greater than that effected wild-type Ad3 12S cDNAs expression. Significantly greater levels of positive transactivation were observed in cotransfections with 13S cDNAs of Ad5 or with the 13S E1A cDNA of Ad3hr15 revertants, compared to the transactivation observed with the mutant encoded wild-type Ad3 13S E1A cDNA. The Ad5 helper and dI-revertant phenotype of Ad3hr15 appear to be related to transactivation activities of coexpressed E1A genes. The nonconserved region which separates the conserved coding regions CR2 and CR3 of the type 3 E1A gene acts to attenuate E1A-mediated repression and transactivation of transcription. PMID- 7491792 TI - The proposed gene for VP1 of HAV encodes for a larger protein than that observed in HAV-infected cells and virions. AB - The termini of hepatitis A virus (HAV) mature proteins have been assigned mainly by their homology to other picornaviruses and their apparent electrophoretic mobility; the proposed coding sequence for VP1 is supposed to encompass 900 nucleotides from position 2208 to 3107 of the HAV genome. In order to further characterize this protein, we analyzed the in vitro-and in vivo-synthesized translation products of the putative VP1 gene. cDNA coding for full-length VP1 was cloned under the control of a T7 promoter in pTF7-5; the resulting plasmid (pTF7-5/VP1) was used for both synthesis of RNA to program rabbit reticulocyte lysates and construction of a recombinant vaccinia virus (rvv/T7-VP1). Immunoblot analysis and immunoprecipitation using antisera raised against a synthetic peptide corresponding to amino acids 13 of 33 of VP1 (13-33/VP1) led to identification of a 37-kDa protein in lysates of in vitro translated VP1 and rvv/T7-VP1-infected HFS cells, whereas a 33-kDa protein was detected with purified virions and in lysates of HAV-infected HFS cells. Because the antiserum used was directed against an amino-terminal part of VP1 and the amino terminus of VP1 is identified by sequence analysis, these results show that VP1 present in the HAV virions and infected cells is shorter than previously proposed and suggest that the real carboxy terminus of VP1 is approximately 40 amino acids upstream. In order to limit the possible carboxy-terminal sites in the predicted region, we investigated in vitro synthesized translation products of a set of constructs with C-termini ending at potential cleavage sites for viral proteases 3C. The construct containing the nucleotides from position 2208 to 3026 codes for a protein (1-273/VP1) which exhibits the same electrophoretic mobility as VP1 synthesized by HAV in vivo. PMID- 7491793 TI - Inhibition of viral aphid transmission by the N-terminus of the maize dwarf mosaic virus coat protein. AB - Since removal of the exposed N-terminus of the coat protein of some potyviruses abolishes aphid transmission, the role of this coat protein region of maize dwarf mosaic potyvirus (MDMV) in aphid transmission was investigated. The viral cDNA encoding this region was cloned and expressed as a fusion protein in bacteria. The resulting purified N-terminus of the coat protein was used in controlled aphid transmission experiments in competition with MDMV. The results show that this region inhibits aphid transmission of MDMV, indicating a direct involvement of the N-terminal region of the coat protein in aphid transmission. PMID- 7491794 TI - Analysis of human immunodeficiency virus type 1 integrase mutants. PMID- 7491795 TI - [Thermo-elastic stress analysis of human bones]. AB - The Thermoelastic Stress Analysis (THESA) is a widely used procedure in motorcar- and airplane engineering. This study investigated the reliability of THESA for stress analysis of human bone. A human femur was cyclic stressed and the resulting stress pattern was scanned from the surface of the bone by means of the thermoelastic stress measuring instrument SPATE 9000. To proof whether the scan of SPATE 9000 is equivalent to the stress distribution of human femur surface, strain gauges are used to control the results at two different regions of the femur diaphysis under equal but static conditions. It could be shown, that both measuring methods lead to corresponding results of stress pattern on human femur surface. PMID- 7491796 TI - [Implant fixation strength and osseointegration following systemic administration of recombinant factor XIII and factor XIII concentrate. Animal experiment with implant fixation strength and osseointegration of porous surface implants]. AB - 30 cylindrical commercially pure titanium fiber porous coated Ti6A14V implants were inserted press-fit into the proximal humeral portion of 30 sheep humeri to determine the systemic effect of recombinant factor XIII and placenta-derived factor XIII concentrate on bone ingrowth and on strength of fixation. For both the recombinant factor XIII and the factor XIII concentrate group the volume fraction of bone ingrowth and the strength of fixation was higher when compared with the control specimens. However the difference was only significant for the factor XIII concentrate group. PMID- 7491797 TI - [Treatment results of modified Glorion-Rideau release in Duchenne muscular dystrophy]. AB - Although the cause of Duchenne muscular dystrophy has been recently found, there is no causal treatment to alter the natural course of this disease. Based on the recommendations by Glorion and Rideau with early treatment of contractures of the hips and the lower limbs we performed a modified release of the spina muscles, resection of tensor fasciae latae and a lengthening of the tendo calcaneus in 32 patients. The mean ae of DMD patients at time of operation was 6.1 yrs. The mean follow-up was 3.4 yrs. All children underwent mobilization the day after surgery. Complete correction of all contractures was immediately achieved after operation and kept in all but two cases up to the follow-up examination. No loss of ambulation was observed. Our results demonstrate that early selective surgery in DMD patients before or just at the onset of contractures without performing an additional aponeurectomy of the iliotibial band and percutaneous tenotomy of the hamstrings according to the original Glorion-Rideau-technique safely prevents severe contractures and should prolong ambulation. PMID- 7491799 TI - [Value of sonographically guided biopsy in the histological diagnosis of benign and malignant soft-tissue and bone tumors]. AB - In 41 patients, an ultrasound guided needle biopsy for histological diagnosis of benign and malignant soft tissue and bone tumors was carried out. In 32 cases following ultrasound guided needle biopsy of the soft tissue component of tumor a final diagnosis could be histopathologically determined; in 5 additional cases, due to the small amount of biopsy, a tumor grouping was able to be ascertained, a further tumor differentiation however could not be made. In 4 cases with cystoid tumors it was not possible to make a diagnosis. The deciding advantage of the new method of ultrasound guided needle biopsy presented here, in comparison to the so called blind tumor biopsy, is that by way of a picture control on the ultrasound screen exact positioning of the biopsy needle is possible. The ultrasound monitor enables one to observe the biopsy procedure, various biopsies from different sites of the soft tissue components of the tumor are possible through one biopsy canal. The new method of the ultrasound guided needle biopsy of benign and malignant soft tissue and bone tumors should however only be carried out in tumor centers. PMID- 7491798 TI - [A new punch technique of cartilage-bone tissue for morphological studies in orthopedics]. AB - A new technique is presented for the analysis of tissues which allows an exact, detailed comparison of light and electron microscopy. For this purpose, the previous techniques required the cutting of tissue pieces from the original tissue compound and the preparation of drawings as a rough localisation of the tissue pieces removed for analysis. Due to the processes of cutting and drawing, only an approximate localisation of the tissue investigated by electron microscopy was practicable. Specifically, the exact analysis of fetal cartilage bone-tissue was mostly not possible due to the complicate arrangement of both tissue components. Therefore, the assessment of electron microscopic analyses was severely limited. The new technique, presented in this study, is characterised by a removal of punches 1 mm in diameter from the original tissue for electron microscopy. The remaining tissue with its exact defined punch-punctures is prepared for standard histology. Thus, the specific advantage of this new method is the precise attribution of the punched tissue to the original tissue group. Human fetal cartilage-bone-tissue has been used to demonstrate this punch technique and the specific light- and electron microscopic preparations. PMID- 7491800 TI - [Incidence of recurrence of aneurysmal bone cysts following surgical treatment and adjuvant therapy with phenol]. AB - Between 1966 and 1991, at the Department of Orthopaedics 34 patients with a mean age of 15.7 years (range 7-46 years) underwent surgery because of aneurysmatic bone cysts. Mean follow-up was 11 years (range 1-26.3 years). We observed a total rate of recurrence of 11.8%. In a group of 9 patients who received local phenol therapy after curettage and bone transplantation no recurrence was observed. The different surgical and conservative methods of treatment are discussed also with respect to rates of recurrence stated in the literature. PMID- 7491801 TI - [Is physiotherapy useful following surgery for carpal tunnel syndrome?]. AB - A prospective follow-up-study of 97 patients operated because of a carpal tunnel syndrome was carried out to investigate postoperative use of physiotherapy. Postoperative 32 patients were treated at least 12 times and 65 patients were not treated physiotherapeuticly. Physiotherapists postoperatively treated by activating joints of fingers and hand. We examined the patients one day preoperative and on an average of 9 months postoperative. Complaints of the patients, local findings, vigorimetrically measured strength and electromyography were registered. The vigorimetrically measured strength of the operated hand improved significantly less in patients treated physiotherapeutically compared to not treated patients. All other complaints and findings improved equally. We interpreted this difference as a result of spare caused by awareness of a sick hand. PMID- 7491802 TI - [Arthroscopic resection of the shoulder joint (Mumford operation)]. AB - Based on results of anatomical studies, we established an arthroscopic procedure to resect all degenerative changes within the joint without endangering the stabilizing structures. Since November 1989 we operated on 26 patients with this technique. There were no major complications. The mean score significantly increased from 64.9 (+/- 12.8) to 86.8 (+/- 11.5). Analysis of postoperative x rays revealed an average length of resection of 21.3 mm at the inferior border of the clavicle, of 15.4 mm in the middle, and of 10.2 mm at the superior border of the clavicle. 21 out of 26 patients were completely satisfied and would undergo the same procedure again. In two of the five unsatisfied patients the resected length of the clavicle was insufficient and these patients required open surgery for resection of the remaining bone. Our first experiences are encouraging. The surgical technique is standardized and reproducible. The short time results score not worse compared to the open conventional technique. With the 2 to 4 year long term follow up we have to prove, whether maintaining the passive stabilizers of the joint improves the long term results better than the open technique. PMID- 7491803 TI - [Stabilization of interligamentous valgus procedure in tibial head osteotomy using staples]. AB - In patients with painful post-traumatic or idiopathic varus gonarthrosis we perform interligamentous valgization osteotomy of the tibial head if arthroscopy shows the lateral joint surfaces to be in sufficiently good condition. Between 1989 and 1993, a total of 75 osteotomies were performed in symptomatic varus gonarthrosis, including 52 cases of idiopathic and 22 cases of post-traumatic gonarthrosis, and one case following infection of the knee joint. The corrections were stabilized exclusively with osteotomy staples (e.g. Richards). The stability thus achieved permits mobilization without plaster with partial loading up to 15 kg from the first postoperative day on. If the indication is adhered to, the operation correctly performed and the complication rate low, we regard the described procedure as appropriate for treating monocondylar varus gonarthrosis. PMID- 7491804 TI - [Ultrasonographic measurement of leg length and leg length differences]. AB - A new diagnostic method for measuring the leg length and the leg length difference by means of ultrasound is presented. A special device for holding and moving the ultrasound transducer was constructed. The measuring points on the hip, knee and upper ankle can be visualised by means of a 5- or 7.5-MHz linear scanner. The measuring device gives the distance of the points in cm so that the difference correspond to the length of the leg, femur and tibia. Tests conducted on corpes and clinical examples show that ultrasound in combination with our special device is an ideal method for determining the exact lengths of the leg. The ultrasound measuring of the leg length offers a reliable, non invasive and easily performed method. Because ultrasound is not limited by radiation hazards, our technique can be used for clinical screening. PMID- 7491805 TI - [Intraoperative assessment of leg length in alloplastic hip joint replacement]. AB - A new measuring device has been developed at the Orthopedic Clinic of the University of Tubingen. Its task is to ensure that the right length of leg is selected intraoperatively. The method of measurement can be used in all approaches for alloplastic hip replacement when the patient is in the supine position. Measurement is quick, non-invasive, and can be carried out in a direct comparison to the contralateral leg, using measuring points at the iliac crest and the upper edge of the patella. In a prospective randomized study we were able to demonstrate on 53 patients that the use of this newly-developed measuring device improves the accuracy of the length of the operated leg as compared to the contralateral leg, so that the difference is only +/- 0.5 cm (post-operative difference in length of leg without use of the measuring device 1.1 cm on average, with the measuring device 0.5 cm; p < 0.05). PMID- 7491806 TI - [Hip pain as only symptom of extensive phlebothrombosis]. AB - We report a case of a 17 year old girl that had 10 years ago a Perthes disease involving her left hip, but complained now of pain and limitation of movement in her left hip. She was febrile and had a blood sedimentation rate of 102/138. Aspiration of her hip was negative. She was a non smoker and was not on oral contraceptive. The left hip showed a painful flexion contracture of 40 degrees. Phlebography revealed thrombosis of her left femoral vein, the common iliac veins and the inferior vena cava. Because of the serious life threatening nature of the condition we feel that it should enter into the differential diagnosis of all orthopaedic surgeons encountering cases with similar presentation in the future. PMID- 7491807 TI - [Histological and morphometric studies of femurs with stable hip joint replacement. An autopsy study with special reference to factors leading to late loosening]. AB - To gain insight into the tissue reactions leading to non-infectious loosening 25 autopsy specimens of femurs with well-fixed cemented femoral components of hip prostheses were analysed histomorphologically and morphometrically. The implant duration ranged from one month to 15 years. With the exception of some focal bone cement contacts bone and cement were separated by a fibrohistiocytic soft tissue membrane. This membrane contained wear particles predominantly of the bone cement mantle and--less abundant--of the polyethylene cups. The amount of wear particles as well as histiocytes and necroses within the membrane statistically significantly increased with advancing time in situ. The cancellous bone adjacent to the soft tissue membrane often revealed a mainly histiocytic infiltration too, accompanied by bone remodeling in this area. The histiocytic inflammatory reaction at the bone cement interface, which is mainly caused by the accumulation of wear particles is considered to be of major importance for the non infectious late loosening of cemented prostheses. PMID- 7491808 TI - [Osteoradionecrosis of the hip joint]. AB - The radiation induced osteonecrosis of the hip is a rare complication after radiation therapy. The time of radiological manifestation is dramatically longer (2 to 17 years) than reported from other bones. There is an inverse correlation between age at the time of radiotherapy and the latency time. Sclerosis, focal lyses, fractures and migration of the femoral head are seen in the radiographs. Differential diagnosis to neoplastic changes should be achieved by bone scan and MRI. We have seen five patients with this condition and had operated on four of them. Treated with THR the fractured acetabulae healed within 9 months and the sclerosis diminished. We did not see any progression of the damage induced by irradiation. PMID- 7491809 TI - [Neuroradiographic diagnosis of degenerative disk diseases--the Dusseldorf Diagnostic Disk program. III. Reproduction of pain and instability]. AB - The goal of this study was to examine the relationship between pain reproduction test and instability in order to point out the clinical significance of radiological instability. The pain reproduction test was correlated with segmental instability of 103 patients and with traction spurs of 172 patients. There was no statistical correlation with the mentioned parameter. Therefore instability is considered only as a morphological parameter but it has no clinical importance. PMID- 7491810 TI - Human schistosomiasis: potential consequences of vaccination. AB - The potential outcomes of a Phase II schistosomiasis vaccine trial are explored using mathematical models which assume some natural immunity and a vaccine providing partial protection for a limited time. Analyses suggest that vaccination may have only a limited impact on life-long cumulative worm burdens and may lead to increased susceptibility and higher worm burdens at some ages, a "rebound" effect which results reduced opportunity to acquire natural immunity. There is a trade-off between the degree and duration of vaccine-induced protection; long-lived vaccines may be preferable to vaccines giving greater protection but for a shorter time. PMID- 7491811 TI - Plant virus expressing hybrid coat protein with added murine epitope elicits autoantibody response. AB - A modified tobacco mosaic virus (TMV) was used to direct the synthesis of TMV coat protein hybrids containing a 13 amino acid sequence of the murine zona pellucida ZP3 protein. This hybrid protein was found to be robust and accumulate to high concentrations in inoculated plants. Virus-like particles containing the hybrid coat protein could be readily purified from infected plant tissue. Parenteral immunization with these virus-like particles as antigen resulted in serum antibody recognizing ZP3 epitope in C57BL/6J and BALB/cBy mice. The antibodies recognized synthetic ZP3 peptide and the authentic murine ZP3 glycoprotein. Antibodies were recruited in vivo to the zone pellucida in female mice. PMID- 7491812 TI - Thermostabilization of live virus vaccines by heavy water (D2O). AB - Eradication of poliomyelitis is based on the mass administration of oral poliovirus vaccine (OPV). Delivery of effective vaccines in the developing world, especially in tropical areas, is compromised when refrigeration cannot be assured. The OPV, prepared with three live attenuated polioviruses (Sabin strains, serotypes 1, 2 and 3), is considered to be the most thermolabile of vaccines in the World Health Organization's Expanded programme on Immunization. To be effective, the initial concentration (potency of each of the three component serotypes, measured in tissue culture infective doses, should not decrease by more than 0.5 log10 before vaccine delivery. High concentration (1 M) of MgCl2 is currently used as stabilizer for OPV. The stabilizing effect of D2O was tested here on OPV strains. By diluting the viral suspension with D2O-based salt and buffer solutions, in a manner similar to that involved in OPV production an 87% concentration of D2O in the final viral preparation was achieved. In severe conditions of testing (incubation for 3 days at 45 degrees C), the Sabin 3 virus lost an average of 2.7 log10 potency in the presence of 87% D2 as compared to 3.0 log10 in H2O-based 1 M MgCl2, and to 5.7 log10 in the H2O-based control solutions. When tested in a combined 87% D2O and 1 M MgCl2 treatment, the Sabin 3 virus lost only 1.3 log10 potency after 3 days at 45 degrees C. Similar thermostabilizing effects were obtained for Sabin 1 and Sabin 2 strains, but the level of stabilization was slightly lower. Tested in standard conditions at 37 degrees C for 7 days, the infectivity of the three D2O MgCl2-treated OPV strains remained in the limit of requirements ( < or = 0.5 log10). The stabilizing effect of D2O was also demonstrated on yellow fever 17D vaccine virus strain. PMID- 7491813 TI - Quantification of whole virus particles (146S) of foot-and-mouth disease virus in the presence of virus subunits (12S), using monoclonal antibodies in a sandwich ELISA. AB - This paper describes a method for the specific quantification of whole virions of foot-and-mouth disease (146S) in the presence of virus subunits (12S). The method involves the use of virus neutralising monoclonal antibodies directed against a linear epitope of the VP1 loop region of a type O virus. The monoclonal antibodies were used as both capture and detecting reagents (labelled with horse radish peroxidase) in a sandwich ELISA. Such monoclonal antibodies also have the advantage that they do not detect viruses containing proteolytically cleaved VP1, thus the assay system is ideal for estimation of whole particles in vaccine manufacture where the immunogenicity of the vaccine depends on virus integrity (whole virions being present) and uncleaved capsid protein. VP1. Other combinations of different anti-type O FMD virus monoclonal antibodies used as capture and detecting reagents were also examined. The system could be adapted to on-line continuous testing of virus being produced during a manufacturing run allowing maximisation of virus yield and quality control. PMID- 7491814 TI - Demonstration of Borna disease virus RNA in peripheral blood mononuclear cells from healthy horses in Japan. AB - Borna disease (BD) is a progressive poliomeningoencephalomyelitis which occurs naturally in horses and sheep. Here, peripheral blood mononuclear cells (PBMC) derived from 57 healthy horses in Japan were examined by a nested reverse transcription-polymerase chain reaction to determine the prevalence of BD virus (BDV) infection. Seventeen (29.8%) of the samples were positive by this examination and the specificity of the amplified product was confirmed by hybridization with authentic oligomer probes. About 60% of the BDV RNA-positive individuals also showed seropositivity by Western blotting. This report is the first for the demonstration of BDV RNA in PBMC of healthy horses, as well as the first on the BDV infection in horses in Japan. Thus, BDV may be more widespread in healthy horses over the world as well as in Japan and the detection of BDV RNA in PBMC at a high rate indicates that the disease might develop in a part of the carriers only after long-incubation period. PMID- 7491815 TI - Preclinical evaluation of an ALVAC (canarypox)--human cytomegalovirus glycoprotein B vaccine candidate. AB - Successful vaccination against the human cytomegalovirus (HCMV) requires induction of both neutralizing antibody and cytotoxic T lymphocyte (CTL) responses. The HCMV glycoprotein B (gB, UL55) would be one of the most important immunogens to induce neutralizing antibodies. We tested the immunogenicity of an ALVAC (canarypox)-HCMV-gB (ALVAC-gB) recombinant in mice and guinea pigs in order to provide preclinical data for a phase I clinical trial of a HCMV vaccine candidate. ALVAC is an attenuated vaccine strain of canarypox virus which replicates productively in avian species but abortively in mammalian cells. The ALVAC-gB recombinant inoculated subcutaneously in mice and intramuscularly in guinea pigs induced HCMV-specific neutralizing antibodies and gB-specific CTL responses. Ultraviolet irradiation of the ALVAC-gB recombinant before immunization diminished CTL responses, indicating that intracellular expression and processing of gB-protein were necessary for CTL induction. Prior immunity to vaccinia virus did not decrease immunogenicity of the ALVAC-gB recombinant in mice. Thus, despite its host range restriction, ALVAC-gB is potentially capable of inducing both humoral and cell-mediated immune responses to HCMV in both vaccinia-immune and non-immune individuals. PMID- 7491816 TI - Incomplete protection of hamsters vaccinated with unlipidated OspA from Borrelia burgdorferi infection is associated with low levels of antibody to an epitope defined by mAb LA-2. AB - Efforts to develop a recombinant vaccine for Lyme disease have focused on using the outer surface protein A (OspA) of Borrelia burgdorferi as an immunogen. We evaluated the effectiveness of an unlipidated recombinant OspA as a vaccine in hamsters. This molecule is soluble and can be produced in high yield in Escherichia coli, characteristics that permit simple and relatively low cost production. Vaccination with unlipidated OspA protected a substantial portion of animals--59-79%, depending on the challenge strain and route--against moderate doses of spirochetes delivered either by injection or by bite of infected nymphal ticks (Ixodes scapularis). The instances of vaccine failure were associated with development of low levels of antibody to a particular OspA epitope, one defined by mAb LA-2. At least 50 ng ml-1 of LA-2 equivalent antibody was necessary for protection of hamsters. Lower LA-2 equivalent antibody concentrations occurred in unprotected animals in the presence of high-titered polyclonal antibody to native OspA. A competitive binding assay to quantitate this serum fraction is described that should be of use in monitoring the quality of the antibody response to OspA in vaccine trials. Concentrations of LA-2 equivalent antibody parallel the ability of the serum specimens to inhibit the growth of B. burgdorferi in culture. PMID- 7491818 TI - Addition of immunotherapy with Mycobacterium w vaccine to multi-drug therapy benefits multibacillary leprosy patients. AB - Immunotherapy with a vaccine consisting of autoclaved Mycobacterium w, was given in addition to standard chemotherapy (multidrug therapy (MDT)) to 93 multibacillary (MB) leprosy patients. One hundred and seven patients with similar types of disease served as controls and received MDT + placebo injections. The study was a double-blind randomised trial. On opening the codes, results obtained were in concordance with those in a single-blind trial which has been extensively reported. Bacteriological clearances were significantly more rapid in vaccinated patients (p < 0.03). Thirty-five LL or BL patients with a high bacterial index (BI) of 6 were completely cleared of acid-fast bacilli (AFB) after eight doses of vaccine. Only 8 patients in the control group became bacteriologically negative in the same time period. They all had BIs < 4. Associated with decreasing BI was accelerated clinical regression of lesions after vaccination and lepromin conversion rates of 100% for BB, 71% for BL and 70% for LL. A significant number of immunised patients showed histological improvement (p < 0.004). Thirty-six showed a complete disappearance of dermal granulomas and a picture of non specific infiltration. The vaccine did not precipitate neuritis or deformities; episodes were noted in vaccinated patients as were incidences of Type 2 reaction. The overall improvement was reflected by a shorter duration of treatment and faster release of vaccinated patients. PMID- 7491817 TI - Safety and immunogenicity of the PFP vaccine against respiratory syncytial virus (RSV): the western blot assay aids in distinguishing immune responses of the PFP vaccine from RSV infection. AB - PFP-1 vaccine was evaluated in a randomized, controlled study in 47 RSV seropositive children. Trivalent inactivated influenza virus (TIV) vaccine was the control. Vaccine reactions were monitored, and bloods were obtained before vaccination, 4 weeks after vaccination, and at the end of the RSV season. Respiratory illnesses were evaluated during the outbreak. Neutralizing antibody (Nt Ab) assay to RSV, IgG ELISA to RSV proteins and a Western blot assay were performed. Acute reactions with the PFP vaccine were mild. An early RSV outbreak resulted in infection of 44.4% of the TIV recipients shortly after vaccination. In the PFP vaccine groups, the Nt Ab and ELISA assays did not distinguish between Ab rises due to natural infection versus vaccine; however, the Western blot assay characterized the post-vaccine rises. Two major Western blot profiles were produced: an infection profile (antibodies that recognized the F and G surface glycoproteins and internal proteins) and a vaccine profile (antibodies that recognized only the surface glycoproteins). The PFP vaccinees who were not infected with RSV developed ELISA and Nt Ab responses to the surface glycoproteins that were similar to the TIV vaccines with natural RSV infection. None of the children developed vaccine-enhanced disease. Thus, the PFP-1 vaccine was safe and immunogenic in RSV seropositive children even when vaccine was administered during a RSV outbreak, and the Western blot assay was useful in distinguishing Ab rises caused by RSV infection versus PFP vaccine. PMID- 7491820 TI - Immunological responses and protection against Mycobacterium bovis in calves vaccinated with a low dose of BCG. AB - Groups of calves (20 per group) were vaccinated subcutaneously with a single dose of BCG Pasteur (6 x 10(4) or 6 x 10(6) colony forming units) and two months later, 15 calves from each group were challenged intratracheally with virulent Mycobacterium bovis. Vaccination with either dose of BCG induced significant protection against the development of tuberculous lesions compared to non vaccinated controls. Seven months after BCG vaccination, many of the vaccinated animals which had no lesions and were M. bovis culture-negative at necropsy showed positive reactions for M. bovis in three assays for measuring cellular immune responses (comparative intradermal test, interferon-gamma assay and lymphocyte proliferation assay). This effect was most noticeable in the BCG vaccinated calves which had been challenged with M. bovis rather than in the non challenged animals. Antibody responses to M. bovis were very low or absent in the calves during the study. The kinetics of the interferon-gamma response of peripheral blood lymphocytes cultured in vitro with bovine PPD showed that BCG vaccination induced a rapid rise in the response followed by a sharp decline, while infection with virulent M. bovis resulted in an increase in the interferon gamma response by four weeks after challenge and this response remained high through the study. PMID- 7491819 TI - Antibody and cytotoxic T-lymphocyte responses to a single liposome-associated peptide antigen. AB - The development of peptide-based vaccines that elicit antibody (Ab) and cellular immune responses has been hampered by the lack of highly immunogenic formulations. In this study, we compared the induction of Ab and cytotoxic T lymphocyte (CTL) responses to a peptide derived from the V3 loop of HIV-1 gp120 (P18 and its cysteine-glycine derivative (CG-P18)) when incorporated into liposomes with lipid A (LA) or mixed with aluminum hydroxide. P18-specific CTL were only observed with liposomes with LA. P18-specific Ab responses were found with liposomes containing CG-P18 but not P18. Increased surface expression of the former, resulted in enhancement of the Ab response without loss of CTL induction. Thus, the manner in which a peptide is localized can influence the outcome of the response induced by highly immunogenic liposome formulations. PMID- 7491821 TI - Humoral and cellular responses induced by intradermally administered cytokine and conventional adjuvants. AB - Serum antibody responses to the model protein antigen avidin were monitored in sheep following intradermal injection of avidin formulated with a range of commercially available and experimental adjuvants, including muramyl dipeptide (MDP), aluminium hydroxide gel (alum), recombinant ovine interleukin1 beta (rovIL 1 beta), rovIL-1 beta + alum, Quil A + alum or Emulsigen Plus. The highest antibody responses were recorded for animals immunised with avidin in rovIL-1 beta + alum, Quil A + alum or Emulsigen Plus, with moderate responses resulting from use of rovIL-1 beta or alum alone as adjuvants. Lower antibody responses to avidin were recorded when avidin was administered alone or with MDP. Delayed-type hypersensitivity (DTH) responses to avidin indicated that the most pronounced cellular response occurred in animals immunised with rovIL-1 beta + alum. Local cellular changes induced after primary and secondary intradermal injections indicated that distinct patterns of cellular recruitment were induced by the different adjuvants. Avidin with MDP resulted in an elevation of CD4+ T cells in the upper dermis while Emulsigen Plus induced an infiltration of large numbers of neutrophils throughout the dermis and reticular layers. CD4+, CD8+ and gamma delta + T cells increased in number and were found evenly distributed throughout these regions. Alum-based adjuvants resulted in the development of distinct cellular accumulations comprising primarily CD4+ T cells and CD45R + B cells arranged in distinct foci in the reticular layer. These cells were strongly class II positive as were the majority of macrophage like cells surrounding the foci. Staining for factor VIII related antigen indicated the presence of endothelial venules in the T and B cell foci and surrounding tissues. PMID- 7491822 TI - Population growth, resources and health: challenges for public health professionals. AB - Current global developments as regards demographic growth and resource utilization represent one issue most difficult to tackle, in terms of (general and health) policy as well as in human terms. In the 1960s-1970s, the debate was active and led, among other things, to an expansion of technical cooperation programs aiming at fertility reduction. The 1974 World Population Conference in Bucharest saw controversy about the respective importance of family planning services, on the one hand, and overall development on the other. The debate was then less visible but has recently been given new impetus, in particular at the September 1994 Cairo International Conference on Population and Development. We have a serious, and in some ways seemingly unmanageable problem, marked by the political difficulties and social constraints which are bound to result from effective actions leading to reduction of resource consumption, or to fertility decrease. Public health professionals may make a significant contribution in promoting better information and dialogue and the view that health has to be an ecosustainable state. PMID- 7491823 TI - [Sleep disorders in professional man and women and their relationship to the workplace]. AB - In Switzerland as well as in many other industrialized countries, sleep disturbances are among the most prevalent health problems. Reliable knowledge about the social and personal conditions of sleep disturbances and especially about their association with conditions is however limited. Using data from the Bern workplace Health Project ("Harz-AsProjekt", n = 774) a logistic regression model was applied to study the association between an indicator of sleep disturbances composed both of problems falling asleep and problems waking-up during the night and several work-related factors. Sleep disturbances were found to be directly associated with perceived work load, time pressure and physical work demands and inversely associated with social acceptance at workplace, work satisfaction and the ability not to become immersed into work. Under comparable working conditions these associations did not differ for men and women. It was concluded that psychosocial demands and resources of the workplace do affect the quality of sleep. PMID- 7491824 TI - [Partnership and sex behavior of consumers of illegal drugs: a survey of 654 persons in the "open" drug scene in Zurich]. AB - In the spring of 1991, a survey was carried out in the "open" drug scene in Zurich's Platzspitz area with the objective of studying drug users' behaviour with regard to relationships and sexuality, a random sample of 654 users (both male and female) of illegal drugs being interviewed and the data drawn from these interviews being compared with corresponding data on average young adults aged 17 to 30. The latter data have been continuously collected since 1986 to evaluate Switzerland's strategy to prevent AIDS. About fifty per cent of drug users are in steady relationships, which is a smaller proportion than among average young adults. On the other hand, occasional sexual contacts (whether or not a steady relationship exists) are three times as frequent among drug users. The use of condoms is slightly more frequent here than among the average population, but is not as yet-and especially among those who are HIV-positive-consistent enough by any means. The influence of drug abuse on sexuality is usually felt to be negative (decline in libidinal energy, impotence), and often brings about a decrease in sexual contacts. This is equally true of both opiates and stimulants (cocaine, amphetamine). Promiscuous behaviour is rare as such and does not occur with any greater frequency than among the average population. However, a large proportion of drug users is at some stage exposed to prostitution. In summary, one can say that the preventive strategy continues to be deficient where the sexual activities of drug users are concerned. A greater effort should be made, in this context, to target the following groups: persons already frequenting advice centers, persons who are HIV-positive, and male prostitutes. PMID- 7491825 TI - [AIDS prevention in European prisons]. AB - Informations on HIV/AIDS in prisons which include 32 prison-systems in 17 European countries have been collected. The principle of equivalence of preventive measures and health care between prisons and the outside community, prison authorities should strive to prevent HIV transmission in prisons. There have been major developments in effective prevention programmes in the community, there has been less progress within the prison-systems. Information on AIDS is provided to prisoners in almost all prison-systems. While condom distribution is practiced in a growing number of systems, no prison systems have adopted syringes/needle distribution or exchange during incarceration. A few system have accepted distribution of a disinfectant (diluted bleach) with instruction on cleaning injection materials. Despite clear recommendations made by international bodies in 1987/88, it must be concluded that discrimination and segregation (restriction from access to workshops, specially work in kitchens) remain widespread in 1992. PMID- 7491826 TI - [Confounding of occupational cancer risk in epidemiological studies due to ignorance of smoking data as exemplified by bladder and lung cancer in painters]. AB - Studies on occupational risk factors for cancer are frequently carried out without information on smoking habits. However, the proportion of smokers may vary considerably across occupational groups. Thus it is unclear, whether an observed increased cancer risk in an occupational group is at least partly the result of a higher proportion of smokers in that group. The paper demonstrates the possible extent of such confounding by using hypothetical and empirical examples of studies concerning the lung cancer and bladder cancer risk in painters. A formula for the indirect control of such confounding is applied. Relative risks greater than 1.2 for bladder cancer and 1.3 for lung cancer should thus not be considered solely the result of confounding. The degree of confounding is less than generally assumed. However, the evaluation of this degree remains questionable in case of an interaction between smoking and occupational exposure. PMID- 7491828 TI - [Drug production: quality status in Greece]. PMID- 7491827 TI - [Need for control in the workplace and motivation for alcohol consumption: a written survey in 3 public civil service agencies]. AB - The links between the need for control (inability to withdraw from work obligations and work commitment) at the workplace and the motivation to consume alcohol is investigated on the basis of 427 male employees in three public administrations or spatially and organizationally homogenous parts of these administrations. By analogy with the work of McClelland and with reference to the concept of the need for control it is conjectured that a link exists between the need for control and the motivation to consume alcohol as a means of coping with stress. Using a linear-additive regression model it becomes evident that the inability to withdraw from work obligations though not the work commitment have an effect on the consumption motive. This effect is not eliminated by the tendency for social desirability and the salary grouping, which are included in the regression model as predictors. In summary, the data proves a link between the inability to withdraw from work obligations on the one side and the consumption of alcohol as a means of coping with stress on the other. PMID- 7491829 TI - Social paediatrics: an integral part of the training for general practitioner in The Netherlands. PMID- 7491830 TI - [Pilot study NFP 1A: how many study participants can be found again after 12 years?]. PMID- 7491831 TI - [Transdermal and oral hormone substitution with estrogens: a comparison]. AB - Application of oral estrone/estrone sulfate and estradiol leads in the circulation to a similar estradiol/estrone-ratio which is below one using oral estrogen while using transdermal estradiol the estradiol/estrone-ratio is greater than one. Overall, transdermal application leads to higher bioavailability of estradiol compared with the oral use of estrone/estrone sulfate or estradiol. Oral estrogens increase hepatic protein synthesis (i. e. SHBG) which is not the case using transdermal application. In addition, by this the bioavailability will be increased. The effect on clinical symptoms is similar and the same is true for the effect on bone. Regarding the lipid metabolism elevation of HDL and thereby improvement of the HDL/LDL-ratio by oral estrogens is obvious. However, triglycerides will be elevated. Overall, both forms of application of estrogens have a positive effect on the cardiovascular system. Different effects on IGF-1 and growth hormone raises the question of the effect on oncogenesis. PMID- 7491832 TI - [Clinical effectiveness of a new matrix plaster: an overview]. PMID- 7491833 TI - [Hormone replacement therapy and the cardiovascular system: on the role of hemostasis]. AB - Postmenopausal women using hormone replacement therapy are likely to experience the first signs of cardiovascular disease at a higher age than women with a comparable risk-profile who choose not to use hormone replacement therapy. Epidemiologic as well as clinical evidence suggest that this benefit prevails even in risk groups such as hypercholsterinemic, hypertensive or diabetic women. Thus, hormone replacement therapy exerts its clinical benefit via steroidal effects on various metabolic pathways. We have reviewed these metabolic effects with particular reference to the differential impact of oral versus transdermal therapy. We have focused on the endothelial function as the common final pathway of all pathomechanisms associated with cardiovascular disease. Endothelial dysfunction may be assessed by analysis of the hemostatic system. Analysis of the available data on hemostatic effects reveals only minor differences between oral and transdermal therapy and gives no support to the notion that only oral hormone replacement therapy provides benefits with regard to cardiovascular disease. PMID- 7491835 TI - [Comparison of transdermal with oral hormone substitution: a multicenter study with a new matrix patch]. AB - 166 postmenopausal patients, aged 45-65 years at baseline, with moderate to severe climacteric symptoms were randomly allocated to an open-label, multi center study which compared the efficacy, safety and acceptance of a transdermal 17 beta-estradiol matrix patch with an oral form of estrogen replacement therapy. In a cyclic sequential regimen, the transdermal system delivered 0.05 mg of estradiol/day. Oral dosages of conjugated equine estrogens were 0.625 mg/day. An oral progestin was also given for 11 days in each cycle for each group. A statistically significant reduction compared to baseline in the primary efficacy parameter, the mean number of hot flashes, occurred with a decrease in each group from 6 per day at baseline to 1 per day at 12 weeks; there was no statistically significant difference between the two groups. The incidence and severity of other postmenopausal symptoms, particularly sweating, difficulty in concentration and palpitations were reduced to a greater extent in the patch group without revealing a significant inter-group difference in the total symptom score. There were also no statistically significant differences in the mean serum estradiol and FSH concentrations between the two treatment groups after 12 weeks of therapy. A similar number of adverse events was observed in both groups. The most frequent adverse events were breast pain and under oral estrogen therapy, gastrointestinal complaints and weight increase. Skin irritation or dermatitis occurred infrequently in the patch group. In summary, the matrix patch represents at least as effective a therapy for postmenopausal symptoms as a standard oral estrogen. PMID- 7491834 TI - [Value of transdermal hormone substitution for postmenopausal osteoporosis]. AB - The osteoporosis is a major public health problem occurring primarily among the postmenopausal women. The relationship to the loss of ovarian function is undoubtful. The consequence of this knowledge is to substitute the hormone deficiency. Hormone replacement therapy (HRT) is effective with oral, subcutaneous, parenteral, and transdermal application. The transdermal hormone replacement therapy for prevention and therapy (early phase) of postmenopausal osteoporosis is effective and sure. The osteoprotective effect was demonstrated in multiple studies. The effectiveness of the HRT can be demonstrated by osteodensitometry and measuring markers of bone resorption. PMID- 7491836 TI - [Kinetics of a new patch for transdermal administration of 17 beta-estradiol]. AB - A novel patch containing 17 beta-Estradiol exhibits improved kinetic profiles compared to the currently available leading transdermal product. The blood concentrations produced by the newly developed matrix patch are stable over 3 to 4 days, thus avoiding the occurrence of 17 beta-Estradiol peaks in the blood. In an additional clinical study an almost linear relationship could be identified between the patch size (Test patch: 7.25, 14.5 and 29.0 cm2) and the obtained 17 estradiol bioavailability (judged on AUC, cmax, c(ave), Cmin). These results are corroborated by the additional in vitro experiments. An almost constant drug delivery rate of 48 micrograms +/- 15 micrograms/day of 17 beta-Estradiol per 13.85 cm2 patch over 4 days can be detected through excised human skin. No statistically significantly different transdermal flux rates of 17 beta-Estradiol were detected in 3 different batches of the transdermal drug delivery system in vitro. Statistical evaluations were performed with the 3-Way-Anova test on the 0.05 significance level. This newly developed product presents a kinetically optimized transdermal 17 beta-estradiol patch for hormone substitution therapy. PMID- 7491838 TI - [Long-term hormone replacement and risk of breast carcinoma: current status. On behalf of the German Menopause Society and the Professional Society of Gynecologic Endocrinology and Reproductive Medicine of the German Society of Gynecology and Obstetrics]. PMID- 7491837 TI - [Estrogen replacement in postmenopause, blood coagulation and fibrinolysis: comparison of a new kind of transdermal estradiol treatment with oral therapy with conjugated estrogens]. AB - Estrogen replacement therapy (ERT) appears to markedly reduce the risk of cardiovascular disease in postmenopausal women. There is evidence that estrogen effects on blood coagulation and fibrinolysis are important mediators of this beneficial effect. It is the acute phase reactants such as factor VII (F VII), von Willebrand factor (vWF) and fibrinogen (Fbg) as well as the main inhibitor of the fibrinolytic system, the plasminogen activator inhibitor (PAI 1), which have been shown to be associated with a particular predisposition or poor prognosis of cardiovascular disease. Additionally, the analysis of stabile reaction products of the coagulation cascade allows for an assessment of the loss of endothelial anticoagulant properties, i.e. endothelial injury. We compared the effects of oral versus transdermal ERT on these key parameters of the hemostatic system. 42 postmenopausal women were randomly assigned to receive either a novel transdermal system releasing 50 micrograms 17-beta-estradiol/24 hours or oral therapy with 0.6 mg conjugated estrogens combined with cyclic medrogestone 5 mg on day 11-21 for three treatment cycles. The study was performed according to the criteria of good clinical practise. We observed no adverse effects on the hemostatic system. Particularly, no increase of coagulatory reaction products, i.e. activity was found. Differences between groups were seen with regard to the extent of favourable effects: While the continuous transdermal ERT significantly reduced factor VII activity, oral ERT had no effect. However, oral ERT significantly reduced PAI 1 concentration by 40% suggesting an improved fibrinolytic capacity.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7491839 TI - Contribution of molecular oncology in the detection of colorectal carcinomas. AB - The development of effective screening tests for colorectal tumors is essential given the high frequency of these cancers in the general population, and more especially in various groups at risk. Sporadic and hereditary colorectal cancers result from the accumulation of mutations in oncogenes, such as ras, myc, neu/HER2, and in tumor suppressor genes such as apc, dcc, p53. The detection of ras or p53 mutations in DNA extracted from stool has been shown to be feasible and might be useful for the development of new screening tests. Many mutations in these genes can also be used as new prognostic factors. Identification of mutation in the apc gene responsible for familial polyposis, or its indirect detection through the study of polymorphism in such families, is completely changing the previously recommended medical attitude for the screening of this disease, and therefore may decrease or even avoid major medical follow-up. These changes are also true for the nonpolyposis hereditary colorectal tumors, also called Lynch syndrome, since the responsible hMSH2, hMLH1, hPMS1 and hPMS2 genes have recently been cloned. Mutations in these genes do not seem to be limited to families with Lynch syndrome, and could account for a predisposition of some patients to develop colorectal or other tumors. PMID- 7491840 TI - Growth factors in colorectal tumorigenesis. AB - Colorectal tumorigenesis involves complex events including multiple genetic alterations in a well-known multistep mechanism called the adenoma-carcinoma sequence. Furthermore, growth and differentiation of normal and transformed cells are influenced by several growth factors, cytokines and hormones. Most of them, like Transforming Growth Factor alpha, Epidermal Growth Factor, Fibroplast Growth Factors, Insulin-like Growth Factors, and gastrin for instance have mitogenic effects. Transforming Growth Factor beta has however a key role in modulating these factors and has preferentially inhibitory growth effect. It is thought to play a strategic role in colorectal tumorigenesis although its mechanisms of action are complex and only partly understood. The aim of the present study is to review the effects of several growth factors, focusing on the role of TGF-beta in colorectal tumorigenesis. PMID- 7491841 TI - Radiology in screening for colorectal cancer. AB - This paper is in two sections. The first reports our own experience at McMaster University with screening for colorectal cancer (CRC), and the second discusses some general aspects of screening, and problems with the methods that are available. PMID- 7491842 TI - Hepatotoxicity of antibiotics. AB - Several antibiotics can cause severe hepatic injury. It is the purpose of this paper to review the main antibiotics that can cause hepatic injury and discuss the presentation, pattern, and outcome of hepatic injury. In the case of the penicillins, the combination amoxycillin-clavulanate and the penicillinase resistant penicillins oxacillin, (di-)cloxacillin, and flucloxacillin can cause (mainly cholestatic) hepatitis. Cephalosporins have little hepatotoxicity; ceftriaxone can cause drug-induced gallstones. The potential of erythromycin and several other macrolides to cause (usually cholestatic) hepatitis is well established. Tetracyclines can cause a syndrome mimicking acute fatty liver of pregnancy, but this complication has virtually disappeared. Quinolones seem to be able to cause cholestasis. Sulfamethoxazole/trimethoprim can cause severe hepatotoxicity, especially in patients with acquired immunodeficiency syndrome (AIDS). Finally, nitrofurantoin can cause acute cholestatic and hepatocellular reactions as well as chronic hepatitis mimicking chronic auto-immune hepatitis. PMID- 7491843 TI - Spontaneous bacterial peritonitis. AB - In this article, authors reviewed etiology, diagnosis, prognosis and treatment of spontaneous bacterial peritonitis. The role of primary prophylaxis is discussed. PMID- 7491844 TI - Clostridium difficile colitis: recent therapeutical and immunological considerations. AB - Clostridium difficile is the main etiological agent of antibiotic associated diarrhoea and pseudomembranous colitis (PMC). It is considered as the most frequent agent of infectious diarrhoea occurring in hospitalized patients, in whom it is responsible for a high morbidity and occasional mortality even when the diagnosis and the treatment are pursued aggressively (1). The pathology is due to the production of at least two toxins: toxin A is an enterotoxin which induces intestinal tissue damage and a fluid response and toxin B is a cytotoxin which lacks any enterotoxic activity but is believed to exert an additive effect in vivo (2). PMID- 7491845 TI - Meckle's diverticulum in association with Crohn's disease. AB - Although it is not uncommon to encounter a Mecklel's diverticulum in a patient with Crohn's disease, the involvement of this diverticulum by Crohn's disease is less common. A well performed radiological small intestinal study strongly suggests the diagnosis, which is definite after histologic examination of the reported. PMID- 7491846 TI - Gastrointestinal leiomyoma and von Recklinghausen's disease: mesenteric angiography demonstrative of a rare association. Report of a case. AB - We report a 51-year-old woman with known neurofibromatosis of von Recklinghausen, who presented twice with melena, caused by a jejunal leiomyoma. The diagnosis was made by arteriography and confirmed by surgery. In order to exclude an intestinal smooth muscle tumour we believe that mesenteric angiography should be the initial procedure of choice in patients with neurofibromatosis presenting with a gastrointestinal bleeding. Genetic analysis suggests that the association of neurofibromatosis and intestinal leiomyoma is more than fortuitous. PMID- 7491847 TI - Paracetamol intoxication and N-acetyl-cysteine treatment. AB - N-acetylcysteine is a good and reasonable specific antidotum in case of paracetamol intoxication. It is very active when administered within eight hours after intoxication. It can be used as well as in intravenous as peroral administration. There are few side effects. PMID- 7491848 TI - Acupuncture in angina pectoris: do psycho-social and neurophysiological factors relate to the effect? AB - We studied the effect of acupuncture in 49 patients with angina pectoris with focus on its relationship to psycho-social factors and changes in skin temperature, pain thresholds, and pain tolerance thresholds. No significant influence from patient expectation, social stress (strain) or profiles of the Minnesota Multiphasic Personality Inventory (MMPI) was found (all p > 0.1). Acupuncture slightly increased exercise tolerance (median 7%), the difference in Systolic Blood Pressure-Heart Rate Product between rest and maximal exercise (delta PRP) (median 3%), and the time to onset of pain (median 10%); decreased nitroglycerin consumption (median 58%) and anginal attack rate (median 38%). Improvement in exercise tolerance was significantly correlated to an improvement in delta PRP (correlation coefficient = 0.7; p < 0.0001) but not to time of myocardial ischemia (correlation coefficient = 0.1; p = 0.1). Compared with 28 patients with a less pronounced anti-anginal effect, the 21 patients with a pronounced effect had a significant increase in local skin temperature, but had no significant change in distant skin temperature and pain thresholds. It is concluded that acupuncture, due to hemodynamic alterations, might have a specific effect on angina pectoris in addition to drug treatments. PMID- 7491849 TI - Clinical and experimental evaluation of cranial TENS in the U.S.: a review. AB - A review of studies evaluating therapeutic applications of cranial TENS in clinical populations reveals frequent use of unreliable self-report outcome measures, insufficient description of treatment protocol, invalid double-blind and placebo conditions and often a lack of adequate description of the electrical parameters used. A review of the experimental literature indicates that cranial TENS is capable of modulating central nervous system activity. It is suggested that the inconclusiveness of the clinical literature may be due to incomplete research design rather than to the questionable nature of the treatment, and that cranial TENS merits continued research. PMID- 7491850 TI - Radiation injury & mercury deposits in internal organs as a result of thallium 201 chloride intravenous injection for SPECT imaging; additional biochemical information obtained in the images of organs from SPECT or PET scans; & potential injury due to radiation exposure during long distance flights. AB - In order to study functional as well as anatomical aspects of various internal organs, SPECT (Single Photon Emission Computerized Tomography) has been used extensively for evaluation of these organs. For SPECT study, intravenous injection of radioactive substances such as technetium-99m (20 millicuries) & thallium-201 chloride (3 millicuries) is commonly used. Although the physical half-life of thallium-201 chloride is 73 hours, its biological half-life is often more than 3.5 times that. Following intravenous injection of thallium-201 chloride it is concentrated in the heart, liver, kidneys, pancreas, thyroid gland, testes or ovaries, and then eventually decays to mercury. Because of its relatively long physical & biological half-lives, thallium-201 chloride may produce mild radiation injury while it remains radioactive. Similar injuries may be induced by technetium-99m (often used for brain SPECT), which radiates Gamma rays (140 KeV), but since its physical half-life is only 6 hours, the side effects are not as significant as those of thallium-201 chloride. Since the main component of thallium-201 chloride radiation is X-ray (68-82 KeV), which consists of photons with a very short wavelength and a high penetrating power, prolonged exposure can induce electromagnetic field-induced injury. As a previous study of the principal author on electromagnetic field exposure indicated, electromagnetic field-induced injury causes the change of L-amino acids to D-amino acids. 2 days after SPECT study of the heart with intravenous injection of thallium-201 chloride, the principal author experienced shortness of breath, loss of appetite, dizziness, fever, and general malaise within the week, and found a progressively significant increase in D-glutamic acid and decrease in L-glutamic acid peaking 2 weeks after the initial injection but lasting for many weeks after in organs such as the heart, liver, kidneys, pancreas, thyroid gland & testes, where radioactive substances had accumulated and radiation was at an average of about 400 counts/min. Even 2 months after the initial injection, the abnormal ratio of D amino acids and L-amino acids had not returned to normal (in the radiation exposed heart, L-amino acids: 6 mg/dl with D-amino acids: 5 mg/dl; normal tissue, L-amino acids: 10 mg/dl with D-amino acids < 1 mg/dl). The principal author tried to find a safe method of reducing possible radiation injury and accelerating the elimination of the already deposited mercury.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 7491851 TI - Role of opioid peptides of rat's nucleus reticularis paragigantocellularis lateralis (RPGL) in acupuncture analgesia. AB - The present study was to investigate the role of the RPGL and its endogenous opioid peptides in acupuncture analgesia (AA) using techniques of brain stimulation and lesion, microinjection, push-pull perfusion and radioimmunoassay (RIA). The results showed that electrical stimulation of the RPGL could increase the pain threshold and enhance the effect of AA, whereas lesion of the RPGL reduce the effect of AA. Microinjection of naloxone (5 micrograms/0.5ul/2min) into the RPGL could partially reverse the effect of AA and the reversal effect of naloxone was dose-dependent. The release of leu-enkephalin (LEK) and B-endorphin (B-EP) from the RPGL in the electrical acupuncture (EA) group was significantly higher than that in the control group (P < 0.05). There were positive correlations between the changes of release of LEK, B-EP and the increase of pain threshold. The results mentioned above imply that the RPGL plays an important role in AA. The activation of the RPGL and the endogenous opioid peptides within it produce an advantageous effect on AA. PMID- 7491852 TI - 'U' technique and standard technique of Norplant removal. AB - OBJECTIVES: To compare the side effects and time required for Norplant removal between the 'U' technique (using the no-scalpel vas deferens holding forceps) and the standard technique. MATERIALS AND METHODS: A randomized controlled clinical trial was carried out of acceptors in family planning clinics in Dr Kariadi Hospital, Semarang, who wanted to have Norplant removal. The Norplant was removed by experienced doctors who had been trained for both the 'U' technique and the standard technique. Removal time, capsule condition and complications were examined. RESULTS: From 41 clients in the 'U' technique group and 41 clients in the standard technique group, the removal times were 2.75 +/- 1.28 and 6.57 +/- 2.93 minutes, respectively (p < 0.01). One week after Norplant removal, complications were found in 5 clients, 1 from the 'U' technique group (2.44%) and 4 from the standard technique group (9.76%). Two weeks after Norplant removal no complications were found. CONCLUSION: The 'U' technique was quicker and less complicated compared with the standard technique. PMID- 7491853 TI - A randomized comparative study of the TCu380A and Cu-Safe 300 IUDs. AB - The Cu-Safe 300 is one of the latest additions to the array of available IUDs. It was specifically designed to decrease unwanted side-effects (e.g. bleeding, pain and expulsion) while providing simplicity of insertion, ease of removal and fair contraceptive protection. For comparison, the TCu380A was chosen. Both types were inserted at random in 600 recipients by a group of independent physicians with a long standing experience in intrauterine contraception. Life table analysis was used for comparison over a period of three years. The Cu-Safe 300 produced a slightly but not statistically significant higher pregnancy rate and more expulsions. Removals for bleeding and pain, however, were significantly less frequent. All inserting physicians agreed that insertion and removal of this new device was remarkably easy. PMID- 7491854 TI - The frameless GyneFix intrauterine implant: a major improvement in efficacy, expulsion and tolerance. AB - The CuFix (GyneFix), conceived in 1985, was developed to minimize three major problems frequently associated with discontinuation of IUD use: expulsion, bleeding and pain. Since the initial clinical investigations, over 10,000 woman years of experience and up to 8 years of follow-up in international multicenter, non-comparative and comparative clinical trials, including a large proportion of nulligravid/nulliparous women, have been collected. Based on new clinical information about the GyneFix from a long-term multicenter clinical trial, conducted in young nulligravid/nulliparous and parous women, the importance of this new contraceptive is discussed. The following conclusions were reached: The unique design characteristics of the GyneFix (frameless, flexible and fixed to the fundus of the uterus) have resulted in optimal tolerance and almost complete absence of expulsion. The result is enhanced effectiveness (comparable to OCs and male/female sterilization) and a high rate of continued use. The GyneFix reduces the IUD failure rate to a minimum and is, therefore, a welcome reversible alternative to OCs and female surgical contraception. Framelessness and flexibility explain the absence of side-effects and adverse events caused by dimensional incompatibility between the frame of conventional IUDs and the uterine cavity and may also explain the absence of PID and ectopic pregnancies in any of the clinical studies. The GyneFix is a promising new, highly effective and safe, contraceptive option for parous women and an equally effective and well accepted method for nulliparous women. PMID- 7491856 TI - Follow-up of women seeking sterilization reversal: a Brazilian experience. AB - OBJECTIVE: To analyze the follow-up of a cohort of women who requested reversal of surgical sterilization. METHODS: This retrospective study was carried out at the State University of Campinas, Brazil. A cohort of 394 women who requested sterilization reversal was followed up for one year after reversal surgery. The analysis includes the percentage of women who completed the steps between the initial consultation up to one year after reversal surgery. RESULTS: After an initial consultation, 55.1% of the patients decided not to continue with the procedures. Of the patients who did undergo laparoscopy (n = 177), 48.6% had no possibility of a successful surgical reversal because the Fallopian tubes were in very poor condition. Only 17.5% of the initial patients requesting reversal ultimately underwent surgery and 3.3% became pregnant. CONCLUSIONS: The analysis of regret about surgical sterilization should include not only women who undergo surgery but also women requesting surgery. Better patient selection and counseling prior to sterilization are necessary in order to prevent increasing rates of sterilization regret in Brazil. PMID- 7491858 TI - Sexual behavior of natural family planning users in Germany and its changes over time. AB - For 10 years, a prospective study has been taking place in Germany to examine the use of natural family planning (NFP). As natural methods are behavioral methods, use-effectiveness, acceptability and continuation rates are very much influenced by patterns of sexual behavior. Therefore we performed an analysis of the sexual behavior of NFP users. Out of the data base of 1211 clients and 12,591 cycles we could identify a group of 300 women, all NFP beginners, with 5900 contraceptive cycles, who contributed at least 12 cycles with reliable recording of their sexual activity. Different groups were analyzed with methods of analysis of variance and regression models to find out significant differences in their sexual behavior with respect to sociodemographic structure and time of use. Nearly half of all the women systematically combine the fertility awareness part of NFP with other family planning methods. They use barriers in more than 60% of their cycles. The other half never or only in about 7% of their cycles use additional barrier methods. The latter show a clear decrease in barrier use in the course of time, whereas the frequent barrier users constantly combine the advantages of two family planning methods. Regarding the frequency of intercourse they are the sexually more active ones and show distinct sociodemographic characteristics. We could confirm the existence of three groups of NFP users, which differ significantly in their use of NFP as a family planning method. Despite these differences the low pregnancy rates indicate the conscious and risk related sexual behavior of the group members. PMID- 7491855 TI - Effects of a progestogen-only pill (Exluton) and an intrauterine device (Multiload Cu250) on breastfeeding. AB - The objective was to compare the effects of Exluton with those of Multiload Cu250, in an open-label, bi-center, randomized study in breastfeeding women. Subjects were randomized to one of the treatment groups 6 weeks after delivery. Women and their infants returned to the study site monthly and follow-up data regarding qualitative and quantitative characteristics of the breast milk, infant growth and infant and maternal health were obtained. There were no statistically significant differences between groups with regard to: (1) amount of milk drunk, amount of milk production, and number of daily breastfeedings; (2) mean milk concentrations of nutritional constituents (except higher milk levels of calcium and magnesium with Exluton during the first month); (3) infant anthropometric parameters. There were no between-group differences with regard to systolic or diastolic blood pressure. During the study, the mean maternal body weight in the Exluton group was significantly lower than in the Multiload Cu250 group. There were no differences between groups with regard to child or maternal health, except a lower incidence of child illness after 6 months in the Exluton group. This study confirms that both Exluton and Multiload Cu250 are acceptable and safe contraceptive methods in breastfeeding women. PMID- 7491857 TI - Natural family planning effectiveness in Belgium. AB - The aim of the study was to check, in a European environment, the practical efficiency and the acceptability of a recent natural family planning (NFP) method. This method includes a double check to detect the beginning as well as the end of the fertile phase of the menstrual cycle. Up to now, 71 women of reproductive age have been registered. They agreed to provide their symptothermal charts and related information. Average age is 32 years. Average fertility is 2 children/woman. Fifty-nine percent of the participants have a professional occupation. The same percentages are found in those accepting a contraceptive method (oral contraceptives or intrauterine devices). Until today 1240 cycles (103 women-years) of experience have been gathered. The average participation was 17.5 cycles/woman. No method failure has been reported. One unintended pregnancy due to unprotected sexual intercourse during the fertile phase occurred (user failure). Overall Pearl Index (PI) was 0.96. In 84% of the cycles, no protected intercourse was reported (NFP only). According to PI, practical efficiency of the NFP method analyzed was 1.31. Sexual abstinence during the fertile phase was found in 71.4% of the cycles. PMID- 7491859 TI - Ovarian function after seven years' use of a levonorgestrel IUD. AB - Fifteen women with regular menstrual periods and seven amenorrheic women who had been using a levonorgestrel-releasing (LNg) IUD for more than seven years were studied. For controls, eight women using TCu380Ag IUDs for more than seven years were studied during two complete menstrual cycles. Ovarian function was assessed with hormonal determination and ultrasound examinations. The regularly menstruating women were studied for two complete menstrual cycles and the amenorrheic women for eight weeks. In the regularly menstruating LNg-IUD users, according to progesterone levels, 93% of the cycles were ovulatory but just 58% of these 'ovulatory' cycles showed normal follicular growth and rupture. Follicular cysts and luteinization of regressing follicles were observed in 42% of the 26 'ovulatory' cycles studied. SHBG capacity was decreased in the LNg-IUD users compared with the TCu380Ag users. Progesterone levels were lower in the LNg IUD users compared with the TCu380Ag users but this difference was not statistically significant. Preovulatory estradiol and LH levels were lower in the LNg-IUD users than in the TCu380Ag users. These differences were not statistically significant. For the amenorrheic women, five had follicular cysts that disappeared spontaneously within 45 days. Two women showed follicular development and rupture. The presence of good cervical mucus was observed in 69% of the ovulatory cycles studied in the LNg-IUD users. This indicates that effects on cervical mucus cannot be the main mechanism of action of the LNg-IUDs. It is concluded that LNg-IUDs may exert a contraceptive effect in many different ways, such as inhibition of ovulation, endometrial changes preventing implantation, alteration of physical and chemical properties of cervical mucus affecting sperm transport and subtle disturbances in hypothalamic pituitary ovarian function, resulting in alterations of follicular development and rupture. PMID- 7491861 TI - Complex study of the physiological role of cadmium. III. Cadmium loading trials on broiler chickens. AB - Cadmium (Cd) loading trials were conducted on a total of 110 (3 x 10 and 4 x 20) broiler chickens prereared for 21 days. The control chickens received no cadmium, while chickens in the six treatment groups were given different doses of Cd as an aqueous solution of CdSO4 administered either into the crop or mixed in the feed. The chickens were kept in a climatized animal house and treated usually for 3-5 weeks (maximum 68 days), with the exception of group Cd-75 chickens which were treated up to 239 days of age. The chickens' health status, body mass and feed consumption were monitored throughout the trial. On days 14-20 and on day 42 of the trial 2 chickens per group, then at the end of trial a total of 25 chickens were killed in anaesthesia. These birds, together with chickens that died or were killed during the trial, were subjected to detailed gross pathological examination. From 11 organs (kidney, liver, spleen, testicle, brain, myocardium, skeletal muscle, lungs, digestive tract, pancreas, tubular bones) of these chickens samples were taken for assay for a total of 16 elements, as well as for light and electron microscopic examination. With the exception of groups Cd-30 and Cd-600, no abnormal clinical signs were observed in the first two weeks of the trial. Chickens of group Cd-30 died before day 8-12 of the trial among signs of complete anorexia, rapid emaciation, huddling and diarrhoea, while chickens of group Cd-600 died before day 28, showing similar clinical signs. The body mass of chickens fed a Cd-supplemented diet either remained constant or decreased substantially, in a degree proportional to the Cd load. The only exception was group Cd-2.5, in which the average body mass of birds at the end of week 8 slightly exceeded that of the controls. Four out of the 10 cockerel chicks fed a diet containing 75 ppm Cd up to 239 days of age died of intercurrent diseases; the remaining six grew well and reached a body mass of 3.8-4.3 kg. Feed conversion efficiency was satisfactory in the control group and in group Cd-2.5 (2.1 and 2.4 kg, respectively) and could not be evaluated in a realistic manner in the other groups. At necropsy, the cockerel chicks of groups Cd-30 and Cd-600 showed severe emaciation, liver and kidney degeneration, myocardial hypertrophy and cardiac dilatation.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 7491860 TI - A cohort study of Norplant implant: side-effects and acceptance. AB - OBJECTIVES: Menstrual irregularity is the main cause of termination of Norplant use; a study of its safety and level of acceptance is needed. STUDY METHODS: A cohort study using medical records of Dr Kariadi Hospital, Semarang, Indonesia and interviews between Norplant and IUD users who had in 1993 been followed-up for 5 years. Expulsion rate, removal rate, level of acceptance and satisfaction were examined. RESULTS: From 170 Norplant and 168 IUD acceptors, the net cumulative pregnancy rates by month 36 of the Norplant and the IUD groups were 0.0% and 1.19%, respectively (p > 0.05). The net cumulative expulsion rates were 0.61% and 1.32% (p > 0.05). The net cumulative rates of removal for medical reasons were 2.37% and 1.97% (p > 0.05). The net cumulative removal rates for non medical reasons were 1.80% and 10.38% (p < 0.001); the net cumulative continuation rates were 95.29% and 85.60% (p < 0.002). The net cumulative acceptance rates were 97.04% and 93.83% (p > 0.05). The Norplant users experienced less menstrual and inter-menstrual bleeding and more amenorrhea than the IUD group. The psychological satisfaction scores among the Norplant and IUD users were high (p > 0.05). The Norplant contraceptive is well accepted. CONCLUSIONS: The findings could be very useful for further steps in promoting Norplant as a contraceptive that is long-acting, with good efficacy, acceptance, and safety. PMID- 7491862 TI - Effect of avoparcin on rumen fermentation and duodenal nutrient flow in sheep. AB - In a digestive-physiological experiment series, the effect exerted by avoparcin on rumen fermentation and on the ruminal digestion of nutrients was studied in wethers provided with rumen and duodenal cannulas, as a function of the composition of feed as substrate. Three control (I, II, III) groups containing different amounts of rumen degradable protein (RDP) and nonstructural carbohydrate (NSC) were formed (composition of diet: group I, 74% RDP and 38% NSC; group II, 57% RDP and 32% NSC; group III, 48% RDP and 23% NSC). The feeding of control diets was followed by the administration of experimental diets containing avoparcin (groups I+A, II+A, and III+A). The dose of avoparcin was 0.75 mg/kg body weight. Irrespective of the RDP and NSC content of the feed, avoparcin reduced the molar ratio of acetic acid and increased that of propionic acid, decreased the acetic acid/propionic acid ratio, and increased the molar ratio of isobutyric acid. Ammonia concentration of the ruminal fluid was significantly lower in group I+A than in the corresponding control group (I). Avoparcin supplementation of diet III improved the apparent digestibility of organic matter from 52.9% to 56.4%. When added to a ration of high NSC and RDP content (I), avoparcin decreased the true digestibility of organic matter from 77.0 to 72.5%. Compared to diet III as well as to diets II and III, avoparcin significantly increased the ruminal degradation of cellulose and hemicellulose, respectively. Avoparcin supplementation of the diet significantly decreased the microbial N content of the duodenal chymus irrespective of the NSC and RDP content of the diet. In group I+A, the amount of dietary N passed from the rumen into the duodenum in 24 h was significantly higher (7.1 g/day vs. 2.7 g/day). In wethers fed the diet of the lowest NSC and RDP content (III), avoparcin supplementation (III+A) increased the apparent digestibility of N in the rumen. In contrast, in wethers fed diets of higher RDP and NSC content (I and II) the true ruminal digestibility of N decreased. Irrespective of the RDP and NSC content of the diet, avoparcin supplementation significantly reduced the efficiency of microbial protein synthesis. The enhanced propionic fermentation induced by the administration of avoparcin allows more efficient utilization of the dietary energy. The higher ratio of undergraded, bypass protein reaching the duodenum provides the animal with a protein source degraded and utilized directly in the small intestine. The results support the observation that avoparcin increases the body weight gain of animals during fattening. PMID- 7491863 TI - Seasonal changes in sperm parameters of British milk rams. AB - Seasonal changes in the sperm parameters of British Milk rams were studied at weekly intervals over a period of 12 months. By comparing the seasonal averages of the test parameters, significant differences were determined in several cases. Substantial differences were found in the averages of quantitative parameters between autumn and the other seasons. As regards the qualitative parameters, the results obtained in summer and autumn were significantly different from those measured in winter and spring. The ratio of deformed spermatozoa was outstandingly high (22.72%) in August. A close negative correlation was established between sperm motility and the incidence of acrosomal defects (r = 0.52; P < 0.1%). The length of natural daylight and daily average temperature were found to exert an influence on the sperm parameters. In autumn, the combined effect of the two weather parameters shows a significant correlation with the concentration of spermatozoa (r = -0.62; P < 0.1%). As British Milk rams show well-balanced sperm production throughout the year with only minor fluctuations, they can be used for reproduction all the year round. PMID- 7491864 TI - Occurrence of Fusarium species and zearalenone in dairy cattle feeds in Vojvodina. AB - The occurrence of Fusarium spp. and zearalenone (F-2 toxin) in dairy cattle feeds was studied during a period of three years (in all seasons) in Vojvodina. Fusarium species were found to be very common in feeds. They were classified into 11 different species (F. chlamydosporum, F. equiseti, F. flocciferum, F. graminearum, F. lateritium, F. moniliforme, F. oxysporum, F. semitectum, F. solani, F. tabacinum and F. tricinctum). Some of these species are known to produce mycotoxins. The highest distribution of Fusarium spp. was observed in the autumn and spring of the second research year, when 53 and 56% of the feed samples, respectively, were contaminated with them. F. oxysporum and F. moniliforme were the most prevalent species throughout the three-year study. Zearalenone was found in various feed samples at concentrations between 140.00 and 960.00 micrograms/kg in all seasons, except in the summer of the first year, in the winter of the second year and in the autumn of the third research year. Pelleted sugar beet pulp and pelleted malt spent grains were the feeds most contaminated with this toxin. Thirty-five out of the 275 feed samples contained zearalenone and 11 of them were contaminated with zearalenone-producing moulds and zearalenone at the same time. PMID- 7491865 TI - Redescription of Goussia neglecta n. comb. (Noller, 1920) (Apicomplexa; Coccidia) and notes on its occurrence in the gut of tadpoles. AB - Coccidian infection in Rana ridibunda and R. esculenta tadpoles was recorded in Hungarian fish farms. Oocysts enclosed in yellow bodies were found in the faeces and in the intestinal epithelium. Developmental stages also had intraepithelial sites. The species was identified with Eimeria neglecta Noller, 1920; however, on the basis of its oocyst morphology it was transferred to the genus Goussia. Besides stages of Goussia neglecta n. comb., intranuclear trophozoites of another coccidian species were also recorded. PMID- 7491866 TI - Effect of exposure to malachite green solution on common carp fry with Dactylogyrus vastator (monogenea) infection. AB - In five experiments, common carp (Cyprinus carpio) fry with Dactylogyrus vastator (Monogenea) infection of different severity, as well as parasite-free fry, were placed into aquaria containing malachite green solution of 0.5-10 mg/l concentration. Most of the fish died within 6 hours. The correlation between the time of death and helminth infection was studied. The experiments demonstrated that in malachite green solution the carp with the heaviest helminth infection died first while the less severely infected specimens survived for the longest time. PMID- 7491867 TI - Prevalence of botfly larvae and lice in studs of North Caucasus (Stawropol County, Russia). AB - From 14 to 17 April 1990 and from 12 to 21 May 1993 a total of 136 Caucasian (Donish, Karabakh, Kabardin, local halfbred) and 26 thoroughbred stud-horses were examined for ectoparasites in studs of Stawropol County, Russia. A total of 4054 botfly larvae were identified from 9700 faecal samples collected during a period of 3-4 days after oral treatment with Eqvalan (MSD), Strongid Plus (Pfizer) and Vermitan Plus (Sanofi-Chinoin). The species distribution was: Gasterophilus intestinalis 84.53%, G. nasalis 10.41%, G. haemorrhoidalis 3.62%, G. inermis 1.41%. Gasterophilus nasalis and G. inermis had not been recorded in the North Caucasian region previously. One hundred and fourteen (of the 136) halfbred and 26 thoroughbred horses were examined for ectoparasites, and all were found to be infested with biting lice (Werneckiella equi equi) and sucking lice (Haematopinus asini). PMID- 7491868 TI - Betamethasone and adult rat lung surfactant lipids. AB - Betamethasone (2 mg/kg/day) was administered to adult male rats (n = 15) for one week by daily subcutaneous injections. Thereafter the lungs were lavaged and lamellar bodies (intracellular surfactant storage granules) were isolated from the tissue. Lipid analyses showed two- to fourfold higher amounts of total phospholipids and disaturated phosphatidylcholine compared with the control. These changes were not found in the kidney liver and were not present in plasma membrane, mitochondrial or microsomal fractions from lungs. It was concluded that glucocorticoids plays a significant effect in increasing the lung surfactant lipid pools of adult rat lungs by changing the phospholipid content of lamellar bodies. PMID- 7491870 TI - Prenatal starvation, betamethasone and lung development in newborn rats. AB - This study examined the potential benefit of simultaneous transplacental betamethasone, which accelerates fetal lung maturation. Pregnant rats were placed in one of 4 groups: Control (C), fed ad libitum until term and given daily physiological saline injections from day 15 of gestation until term; Betamethasone (B), fed as group C but given daily physiological saline injections of 2.0 mg betamethasone/kg body weight from day 15 until term; Starved (S), given 50% rations from day 15 until term and injected as group C; Starved+Betamethasone (SB), fed as group S and injected as group B. Controls and group B did not differ in body or lung weight, protein or DNA, but group B lungs contained more lavageable and tissue surfactant. The S neonates weighed about 40% less than controls, with a proportional reduction in lung weight, DNA, protein or lavage and tissue phospholipids. Betamethasone may alleviate the impact of starvation on the developing lung by accelerating the process of alveolarization which was solved by caloric deprivation. PMID- 7491869 TI - The effect of betamethasone on phosphatidylcholine species composition in fetal rat lungs. AB - After betamethasone treatment, a significant diminution of the monoenoic phosphatidylcholine 32 carbon species (palmitoyl-palmitoleyl phosphatidylcholine and palmitoleyl-palmitoylphosphatidylcholine) and the phosphatidylcholine 34 carbon species (primarily palmitoyl-oleoyl-phosphatidylcholine) could be demonstrated both in absolute and relative terms, while the palmitic acid portion in the phosphatidylcholine fatty acids was nearly unchanged. This is consistent with a significant reduction of the palmitoleic and oleic acid portions of the total phosphatidylcholine fatty acids in the lungs of rat fetuses. PMID- 7491871 TI - Protective effect of verapamil on regional myocardial ischaemic injury in dog. AB - Ischaemic injury was produced in the dog heart by occluding the left anterior descending coronary artery just below the second diagonal branch for 3 h. Verapamil 5.0 mg/kg was injected intravenously 10 min before the occlusion. There was some decrease in ATP and creatine phosphate levels in the hearts of "no drug" animals as compared to sham-operated dogs. Verapamil significantly increase the level of these metabolites. The ATP/ADP ratio decreased from 8.5 +/- 0.60 in hearts from sham-operated dogs to 3.5 +/- 0.30 in the "no drug" group. Verapamil pretreatment resulted in values of the ATP/ADP ratio which were lower than, but not significantly different from, those observed in the control group. In ischaemic mitochondria the calcium content (Ca++) increased significantly as compared to the control and the Verapamil group. A significant increase in free fatty acid content in myocardial membrane preparations was also observed as compared to the sham-operated and the Verapamil group. PMID- 7491872 TI - Toxicological studies on potentiated ionophores in chickens. I. Tolerance study. AB - The tolerance of chickens to monensin (12.5 mg/kg of feed) and maduramicin (3.0 mg/kg of feed) fed at a reduced dose in the presence of the antioxidant duokvin was studied in two experiments including 2 x 200 Tetra-82 broiler chickens. Tolerance was assessed by the appearance of clinical signs indicative of a toxic effect, the number of deaths, the groups' body weight gain, feed and drinking water intake, the aspartate aminotransferase (AST) and lactate dehydrogenase (LDH) activities, calcium ion, inorganic phosphate and total protein content of the blood plasma, the haematocrit value, and haemoglobin concentration. When applied at a dose that had proved to be optimum in the efficacy studies, neither the monensin-duokvin combination (12.5 mg monensin per kg of feed + 120 mg duokvin per kg of feed) nor the maduramicin-duokvin combination (3.0 mg maduramicin per kg of feed + 120 mg duokvin per kg of feed) exerted a statistically significant influence on the parameters tested. PMID- 7491873 TI - Toxicological studies on potentiated ionophores in chickens. II. Compatibility study. AB - Two trials were carried out on a total of 2 x 360 Tetra-82 broiler chickens to study how the presence of the antioxidant duokvin as potentiating agent influenced the compatibility of reduced doses of monensin (12.5 mg/kg of feed) or maduramicin (3.0 mg/kg of feed) with other chemotherapeutic agents (tiamulin, erythromycin, sulfaquinoxaline, sulfachlorpyrazine, flumequine, tylosin, kitasamycin) widely used in broiler rearing. Compatibility was assessed on the basis of the appearance of clinical signs suggestive of toxic interaction, the mortality rate, body mass gain, feed consumption and drinking water intake, and changes in AST and LDH activities of the blood plasma. The monensin-duokvin combination (12.5 mg monensin/kg of feed + 120 mg duokvin/kg of feed) was found to be compatible with erythromycin, sulfaquinoxaline, sulfachlorpyrazine, flumequine, tylosin and kitasamycin. For tiamulin, a slight incompatibility was observed; however, this was much less severe than that found for monensin administered at a dose of 100 mg/kg of feed. The maduramicin-duokvin combination (3.0 mg maduramicin/kg of feed + 120 mg duokvin/kg of feed) was compatible with all the compounds tested; thus, it can be safely applied also in combination with tiamulin. PMID- 7491874 TI - Toxicological studies on potentiated ionophores in chickens. III. Electrotoxicological investigations. AB - The effect exerted by the simultaneous administration of tiamulin (40 mg/kg body mass) and a lowered dose (12 mg/kg of feed) of monensin combined with the antioxidant duokvin on the conductivity of peripheral nerve and the electrophysiological function of the heart was studied in two trials comprising 3 x 6 broiler chickens. For the study of peripheral nerve function, chickens were anaesthetized with pentobarbital sodium, the sciatic nerve (n. ischiadicus) was exposed in the sciatic arc, and bipolar stimulating and efferent electrodes were inserted directly beside the nerve, at an average distance of 20 mm from each other. For the recordings, the nerve was stimulated with 16 supramaximal impulses of 0.02 ms duration, applied at 15-s intervals. The responses given to the stimulus were recorded with the help of a polygraph and evaluated with a computer in online mode. For the study of cardiac function, the chickens were anaesthetized with methomidate. The electrocardiograms were taken in Einthoven's lead II with the help of needle electrodes, and then evaluated with computer in online mode. In control chickens, maximum conduction velocity was 30.70 +/- 0.52 m/s on the average. That value did not change after a single treatment with monensin-duokvin + tiamulin, while it underwent a slight but statistically significant decrease after two treatments. Studies on the electrical function of the heart revealed no signs of dysfunction in either of the treated groups as compared to the control.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7491875 TI - Effects of repeated oral doses of Dikamin D (2,4-D-amine Na) on rats. AB - The toxic effects of repeated, increasing oral doses of Dikamin D (72% 2,4-D amine Na), a broad leave herbicide product used world-wide, were evaluated on rats by the method of Lim et al. (1961). A comparison of the determined acute oral LD50 and the calculated subchronic oral LD50 values revealed a definite tolerance of the experimental animals to the test compound. This finding indicates that repeated oral treatment is capable of increasing the test animal's metabolizing capacity, which accounts for the development of tolerance. PMID- 7491876 TI - Experimental furazolidone toxicosis in broiler chicks: effect of dosage, duration and age upon clinical signs and some blood parameters. AB - Groups of broiler chicks of one day and three weeks of age were given feeds containing 0, 400, 800 and 1000 mg furazolidone (Fz)/kg for three weeks. The age of the birds and the dose and duration of Fz treatment significantly (P < or = 0.05) influenced the parameters studied. Clinical signs of Fz toxicosis included ascites, leg weakness and nervous derangement like convulsions and torticollis. The body weight also decreased. Ascites and nervous derangement was not observed in birds fed 400 mg Fz from one day of age. Fz-fed birds developed anaemia and had lower plasma total protein and albumin levels than those fed no Fz. The decrease in body weight was related to reduced feed intake. Decreases found in the haematological parameters and plasma proteins showed no correlation with the chicks' age but were related to the dose and duration of Fz treatment. PMID- 7491877 TI - Efficacies of dexfenfluramine and fluoxetine in preventing weight gain after smoking cessation. AB - We tested whether 14 wk of dexfenfluramine (30 mg) or fluoxetine (40 mg) treatment would prevent weight gain after subjects quit smoking. Normal-weight women (n = 144) were randomly assigned to drug or placebo on a double-blind basis for 2 wk before quitting smoking and 12 wk thereafter. The fluoxetine group had more dropouts (28/49, 57.1%) than the dexfenfluramine group (17/47, 36.2%), with an intermediate number of dropouts from the placebo group (21/48, 43.8%). All groups gained weight during treatment, but their amount and pattern of weight gain differed. In the first month after quitting smoking, the placebo group gained more weight than either the dexfenfluramine or fluoxetine group (P < 0.05). By 2 mo postcessation, dexfenfluramine still suppressed weight gain in comparison with placebo (P < 0.05); weight gain with fluoxetine was not differentiable from either dexfenfluramine or placebo. By 3 mo postcessation, the dexfenfluramine group had gained 1.0 +/- 0.7 kg, significantly less than either the placebo (3.5 +/- 0.7 kg) or fluoxetine (2.7 +/- 0.5 kg) groups. Three months after drug discontinuation, formerly medicated, but not placebo patients, showed additional weight gain, eliminating differences between groups. Results indicate that weight gain, an adverse accompaniment of smoking cessation, can be minimized to some degree by serotoninergic drugs, although only for the duration of drug treatment. PMID- 7491878 TI - Effect of calcium supplements and stage of lactation on the calcium absorption efficiency of lactating women accustomed to low calcium intakes. AB - The effect of calcium intake on the calcium absorption efficiency from 100 mL cow milk was measured in lactating Gambian mothers habituated to a low-calcium diet [mean intake 7.08 mmol (283 mg)/d], and compared with UK lactating mothers consuming high-calcium diets [mean intake 29.2 mmol (1168 mg)/d] by using a double stable-isotope technique (oral 44Ca and intravenous 42Ca). In a double blind trial starting 9 d postpartum, Gambian mothers were given a calcium supplement [17.85 mmol (714 mg)/d] or placebo for 12 mo. At 3 and 12 mo postpartum, mean (+/- SEM) calcium absorption from isotopically enriched milk was 52.3 +/- 3.1% (n = 25) and 47.2 +/- 4.8% (n = 24) in the unsupplemented Gambian mothers and 48.8 +/- 2.8% (n = 28) and 42.9 +/- 3.7% (n = 24) in the supplemented mothers, respectively. There was no effect of supplementation or stage of lactation on the efficiency of calcium absorption. At 3 mo postpartum the UK mothers absorbed 32.2 +/- 3.8% of the isotopically enriched calcium added to milk, which was significantly less than that of the Gambian mothers (P < 0.01). PMID- 7491879 TI - Differential incorporation of fish-oil eicosapentaenoate and docosahexaenoate into lipids of lipoprotein fractions as related to their glyceryl esterification: a short-term (postprandial) and long-term study in healthy humans. AB - We investigated how the distribution of eicosapentaenoate (EPA, 20:5n-3) and docosahexaenoate (DHA, 22:6n-3) in the sn-2 and sn-1(3) positions of fish-oil triacylglycerols influenced their respective incorporation into triacylglycerol, cholesterol esters, and phospholipids of two lipoprotein fractions: low- and very low-density lipoprotein (VL/LDL) and high-density lipoprotein (HDL). Nine healthy volunteers were studied over both a short-term (0-8 h) and a long-term (30 d) postprandial period of daily supplementation with 2 g EPA and 1.3 g DHA given as 11 g fish-oil triacylglycerol in which DHA was predominantly situated in the sn-2 position. Our results strongly suggest that the higher triacylglycerol incorporation of DHA and the higher metabolic availability of EPA compared with DHA for phospholipid accumulation (particularly in the short-term study) depend on their respective preferential sn-2/sn-1(3) positions in fish-oil triacylglycerol, emphasizing the important role of the triacylglycerol structure and its potential manipulation for modulating availability of either or both fatty acids. PMID- 7491880 TI - Effect of variations in oral fat and carbohydrate load on postprandial lipemia. AB - In this study we assessed the acute effects of the consumption of varying amounts of fat and fructose on the magnitude of postprandial lipemia. Subjects were studied after an overnight fast on four separate mornings, ingesting in random order 5, 40, or 80 g fat, or 5 g fat plus 50 g fructose. Vitamin A (36 mg, or 120,000 U retinol) was also given and blood was drawn at frequent intervals over the next 10 h for measurement of triacylglycerol and retinyl palmitate (RP) concentrations in plasma and the Sf > 400 and Sf 20-400 lipoprotein fractions. (Sf denotes flotation units.) In general, the postprandial triacylglycerol response increased in plasma and in both lipoprotein fractions as a function of both the baseline fasting triacylglycerol concentration and the amount of fat ingested. However, no matter how high the fasting plasma triacylglycerol concentration, there was no increase in the postprandial triacylglycerol concentration in plasma or either lipoprotein fraction after the 5-g oral fat load. The results of the measurements of RP concentration were somewhat similar in that there was a dose-dependent increase in the plasma and the Sf > 400 lipoprotein fraction in response to the higher fat loads. However, just the opposite was true in the Sf 20-400 lipoprotein fraction, for which the increase in RP concentration was inversely related to the size of the fat load.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7491881 TI - Erythrocyte L-aspartyl-L-phenylalanine hydrolase activity and plasma phenylalanine and aspartate concentrations in children consuming diets high in aspartame. AB - A deficit of alpha-aspartyl-phenylalanine (alpha-Asp-Phe) hydrolase activity has been suggested as a cause of possible adverse effects of aspartame ingestion. Twenty-five normal preschool children and 23 school-age children described by their parents as sensitive to sugar were fed diets high in sucrose, aspartame, or saccharin for three successive 3-wk periods. Blood samples were obtained at baseline (fasting) and within the last 3 d of each dietary period (postprandial). alpha-Asp-Phe concentrations were below detection limits (0.5 mumol/L) in all plasma samples and Phe and Asp concentrations remained within normal limits, alpha-Asp-Phe hydrolase activities in baseline hemolysate samples did not differ between groups. One subject had a plasma alpha-Asp-Phe hydrolase activity > 2 SD below the mean. Despite this low activity, this subject did not show consistent cognitive or behavioral anomalies that could be linked to low hydrolase activity. PMID- 7491882 TI - Honey may have a laxative effect on normal subjects because of incomplete fructose absorption. AB - Honey contains fructose in excess of glucose, which may lead to incomplete fructose absorption associated with abdominal symptoms and/or diarrhea. This hypothesis was investigated in 20 healthy volunteers (13 males, 7 females) with a mean (+/- SD) age of 35.9 +/- 12.1 y. Each subject drank the following aqueous solutions in random order: 20 g lactulose, 100 g honey, 50 g honey, and 35 g each of a glucose and fructose mixture. The breath-hydrogen concentration was measured every 15 min for 6 h. Semiquantitative estimates of carbohydrate malabsorption were assessed with lactose as a nonabsorbable standard. Breath-hydrogen concentrations increased by 52 +/- 6, 30 +/- 4, 20 +/- 3, and 4 +/- 1 ppm (mean +/- SEM) after each of the four test solutions, respectively. The estimated carbohydrate malabsorption was 10.3 +/- 1.8, 5.9 +/- 1.2, and 0.5 +/- 0.2 g after 100 g honey, 50 g honey, and the glucose-fructose mixture, respectively (F[2,57] = 16.05, P < 0.001). Within 10 h after the ingestion of 100 g honey, 50 g honey, and the glucose-fructose mixture, six, three and none of the volunteers, respectively, reported loose stools (chi 2 = 7.1, df = 2, P < 0.03). The results of this study suggest that carbohydrate malabsorption after ordinary doses of honey is frequent in healthy adults and may be associated with abdominal complaints. Honey may have a laxative effect in certain otherwise healthy individuals, probably because of incomplete fructose absorption. PMID- 7491884 TI - Antioxidant status in long-term adherents to a strict uncooked vegan diet. AB - Antioxidant status was investigated in 20 Finnish middle-aged female vegans and in one male vegan who were following a strict, uncooked vegan diet ("living food diet"), by means of a dietary survey and biochemical measurements (blood concentrations of vitamins C and E and beta-carotene, and the activities of the zinc/copper-dependent superoxide dismutase and selenium-dependent glutathione peroxidase). Values were compared with those of omnivores matched for sex, age, social status, and residence. Antioxidant supplementation was used by 4 of 20 female vegans and by 11 of 20 control subjects. Based on dietary records, the vegans had significantly higher intakes of beta-carotene, vitamin E, vitamin C, and copper, and a significantly lower intake of selenium than the omnivorous control subjects. The calculated dietary antioxidant intakes by the vegans, expressed as percentages of the US recommended dietary allowances, were as follows: 305% of vitamin C, 247% of vitamin A, 313% of vitamin E, 92% of zinc, 120% of copper, and 49% of selenium. Compared with the omnivores, the vegans had significantly higher blood concentrations of beta-carotene, vitamin C, and vitamin E, as well as higher erythrocyte superoxide dismutase activity. These differences were also seen in pairs who were using no antioxidant supplements. The present data indicate that the "living food diet" provides significantly more dietary antioxidants than does the cooked, omnivorous diet, and that the long term adherents to this diet have a better antioxidant status than do omnivorous control subjects. PMID- 7491883 TI - Effect of different vitamin A intakes on very-low-birth-weight infants. AB - Formula-fed infants with birth weights < or = 1500 g (n = 61) were stratified by 250-g birth-weight ranges and randomly assigned to receive one of three preterm infant formulas (vitamin A contents of 820 IU, 1640 IU, or 2900 IU/MJ; 1 RE = 3.3 IU vitamin A activity) when subjects tolerated 0.314 MJ.kg-1.d-1. Experimental formula feedings were continued until infants weighed approximately 2 kg or until hospital discharge. Vitamin A status as indicated by serum retinol and retinol binding protein (RBP) concentrations significantly decreased during experimental formula feeding at the lowest vitamin A intake. All subjects fed the formula providing the lowest vitamin A intake had hyporetinolemia (< 0.70 mumol/L, or < 20 micrograms/dL), which occurred less frequently (P < 0.05) with the intermediate (6 of 20) and the high (6 of 21) vitamin A intakes. Other outcome measures, including increases in weight, length, and head circumference, and ventilatory support and oxygen therapy, were not different among groups. After the end of the experimental formula-feeding period, all infants were fed standard infant formulas with a vitamin A content of 715 IU/MJ. In a subset of 19 of these infants, subsequent vitamin A status was monitored at ages 6-12 mo and was found to be comparable with that of older children and adults, regardless of the vitamin A content of the formula fed during hospitalization.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7491885 TI - Influence of combined antioxidant nutrient intakes on their plasma concentrations in an elderly population. AB - Data from a cross-sectional survey of 746 non-institutionalized, Boston-area elderly individuals (aged > or = 60 y) were analyzed to assess the relation between antioxidant nutrient intake and plasma antioxidant status. Intakes of vitamin C and carotenoids and supplemental vitamin E were estimated by using 3-d diet records. Mean plasma concentrations of these nutrients were calculated within categories of intake, and polynomial contrasts were used to test for linear trends of the plasma nutrient concentrations across these categories. Adjustments for the corresponding intake of the plasma nutrient under consideration, as well as age, sex, and smoking status were made to minimize potential confounding. Plasma alpha-tocopherol concentrations were 18% greater in individuals consuming > or = 220 mg vitamin C/d compared with those with intakes < 120 mg/d (P for trend < 0.001). Plasma carotenoid concentrations were 13% higher across increasing categories of vitamin C intake (P for trend = 0.002). An increasing intake of carotenoids was moderately associated with higher plasma alpha-tocopherol (P for trend = 0.008) and unrelated to ascorbic acid status. An increasing intake of supplemental vitamin E was weakly correlated with plasma ascorbic acid (P for trend = 0.05) and unrelated to carotenoid status. These results provide epidemiologic evidence that increasing intake of either vitamin C, vitamin E, or carotenoids is associated with greater plasma concentrations of one or both of the other antioxidant vitamins and not associated with any impairment in antioxidant status. PMID- 7491886 TI - Effect of milk and fermented milk on iron absorption in ileostomy subjects. AB - This study was undertaken to determine whether milk with its high calcium content adversely affects the absorption of nonheme iron from the diet as greatly as single-meal studies indicate. Nine ileostomy subjects ate a low-fiber, low phytate diet for 8 consecutive weeks. During the first and eight weeks they drank a 250-mL soft drink with three main meals and an evening snack each day (0.16 g Ca/d). During the two intervening 3-wk periods, they drank the same amount of low fat milk or fermented low-fat milk (Verum; Halsofil, Norrmejerier, Umea, Sweden) according to a formally randomized crossover design (1.4 g Ca/d). During the last 2 d in each of the four periods, apparent iron absorption (balance) from a test diet together with that period's beverage was measured and the plasma ferritin concentration was determined. There was no decrease in apparent iron absorption during the milk diet periods. PMID- 7491887 TI - Calcium bioavailability from a calcium- and sulfate-rich mineral water, compared with milk, in young adult women. AB - Some mineral waters have a high calcium content and may contribute a significant part of the human daily requirements. Calcium bioavailability from a calcium- and sulfate-rich mineral water (CS-W) containing 11.2 mmol Ca/L (467 mg/L) was compared with that from milk in nine healthy young women. Calcium absorption was measured in the fasting state with a dual-label stable-isotope technique. Fractional absorption rates from milk and CS-W were 25.0 +/- 6.7% and 23.8 +/- 4.8% (means +/- SD), respectively, and did not differ significantly (P = 0.05). Urine was collected for 36 h after the administration of the oral stable isotope while the subjects consumed a controlled diet and a quantity of milk or CS-W providing 25 mmol (1000 mg) Ca. No significant difference was found in the excretion of calcium, nor in the excretion of the two stable isotopes. Mean urinary sulfate excretion was significantly increased by 35% when the CS-W was consumed. No significant correlation was found between 36-h urinary excretion of the intravenous calcium tracer and sulfate, sodium, or urine volume. Therefore, calcium from the CS-W was as well absorbed and retained as that from milk, and no calciuric effect of sulfate was found, showing that such mineral waters can be valuable dietary sources of calcium. PMID- 7491888 TI - Oat beta-glucan increases bile acid excretion and a fiber-rich barley fraction increases cholesterol excretion in ileostomy subjects. AB - The purpose of this study was to investigate whether oat beta-glucan is responsible for the increased bile acid excretion previously observed with oat fiber diets. The excretion patterns in ileostomy subjects given diets containing oat-bran bread with and without added beta-glucanase, a beta-glucan-degrading enzyme, were compared. The effect of a beta-glucan-rich barley fraction on sterol excretion was also investigated. Nine ileostomy subjects were served four diets in random order, each diet for 2 consecutive days. Four different kinds of bread, mainly made from oat bran (OB diet, 12.5 g beta-glucan/d), oat bran with beta glucanase (OBE diet, 3.8 g beta-glucan/d), barley (B diet, 13.0 g beta-glucan/d), or wheat flour (W diet, 1.2 g beta-glucan/d) were added to a basal diet. The 24-h excretion of bile acids was 53% higher in the OB diet period than in the OBE diet period (P < 0.05) and also was significantly higher than in the B and W diet periods (P < 0.05). Median (range) bile acid excretion was 851 (232-1550), 463 (123-1414), 755 (133-1187), and 606 (101-980) mg/d in the OB, OBE, B, and W diet periods, respectively. The excretion of cholesterol was significantly higher in the B diet period than in the OBE and W diet periods (P < 0.05), but the mechanism behind this effect of barley fiber is unknown. In oat bran, however, beta-glucan mediates an increase in bile acid excretion, which most probably explains the effect of oat fiber in lowering serum lipids. PMID- 7491890 TI - Effect of green lentils on colonic function, nitrogen balance, and serum lipids in healthy human subjects. AB - Green lentils are an increasingly popular food, but their effects on human colonic function and serum lipids have been studied little. Nine healthy males aged 19-38 y consumed for 3-wk periods a controlled Western diet and the same diet supplemented with 130 g dry lentils/d [which contained 11.8 g non-starch polysaccharide (NSP)] incorporated into loaves, cakes, and soups. Protein was equilibrated with soy protein isolate and carbohydrate with soft drinks. Radioopaque markers were used to calculate mean transit time (MTT) and to correct fecal weight for infrequency of bowel movements. Feces were collected throughout the study and blood was taken on 2 d at the end of each period. Lentils increased fecal weight from 131 +/- 12 g/d (means +/- SEM) to 189 +/- 17.4 g/d (44.9%) (P < 0.005). MTT was unchanged: 46 +/- 6 h for the control diet and 43 +/- 4 h for the lentils (NS). Fecal nitrogen was increased to 2.49 +/- 0.08 g/d for lentils compared with 1.74 +/- 0.09 g/d for the control diet (P < 0.001) and urine nitrogen decreased to 15.31 +/- 0.52 g/d with the lentils compared with 15.90 +/- 0.51 g/d for the control diet (P < 0.05); nitrogen balance was unaffected. Serum lipids were unchanged by addition of lentils to the diet. Green lentils were effective in increasing fecal weight and can therefore make a valuable contribution to a healthy diet. PMID- 7491891 TI - Fatty acid pool size in plasma lipoprotein fractions of cystic fibrosis patients. AB - The objective of this study was to quantitatively measure the essential fatty acid pools in lipid fractions of plasma lipoproteins and the alteration of these pool sizes after consumption of a physiologic intake of fish oil. Twenty-three cystic fibrosis (CF) patients and 21 normal subjects were supplemented with fish oil, providing 35 mg n-3 fatty acids/kg body wt for 4 wk. Blood was collected pre and postsupplementation and was quantitatively analyzed to determine the fatty acid pool size in each lipid class of low-density lipoproteins (LDLs), high density lipoproteins (HDLs), and very-low-density lipoproteins. Two 7-d food records were collected to determine total fat and fatty acid intakes. Intakes of protein, carbohydrate, and fat as a percentage of energy and the ratio of polyunsaturated to saturated fats was similar for the CF and control groups. Energy intake was greater for CF subjects. Smaller lipoprotein essential fatty acid pools were observed in CF patients than in control subjects. Healthy subjects had larger essential fatty acid pool sizes in cholesterol ester fractions of HDLs and LDLs than CF subjects. Cholesterol ester and phosphatidylcholine pools transported the majority of essential fatty acids in both CF and control subjects. PMID- 7491889 TI - Alteration of the fiber and lipid components of a defined-formula diet: effects on stool characteristics, nutrient digestibility, mineral balance, and energy metabolism in humans. AB - Eighteen healthy males with a body weight of 70.0 +/- 3.1 kg consumed three defined-formula diets that varied only in their fiber and/or lipid components: 1) 6.4% fiber (100% soy polysaccharides) and 13.1% lipid [50% medium-chain triacylglycerols (MCTs), 40% corn oil, and 10% soy oil]; 2) 3.4% fiber (75% oat fiber, 17.5% gum arabic, and 7.5% carboxymethylcellulose) and 15.6% lipid (20% MCTs, 50% canola oil, and 30% high oleic acid safflower oil); and 3) 4.4% fiber (same as diet 2) and 14.5% lipid (same as diet 1). Consumption of diet 2 resulted in slightly firmer stools and provided the greatest amount of fecal output per unit fiber intake. Total dietary fiber (TDF) digestibility was lowest for men fed diets 2 and 3, but nitrogen and lipid digestibilities and energy metabolism criteria were not different among diets. Although mineral excretion patterns differed among treatments, fiber and lipid components of the diets appeared not to be responsible for these differences. Results indicate that fecal output can be maintained with a lower intake of a blend of oat fiber, gum arabic, and carboxymethylcellulose compared with soy polysaccharides. Except for TDF digestibility, alteration of amounts and/or sources of fiber and lipid components of defined-formula diets used in this experiment did not alter nutrient digestibility, energy metabolism, or mineral retention. PMID- 7491892 TI - Absorption of dietary quercetin glycosides and quercetin in healthy ileostomy volunteers. AB - Quercetin is a dietary antioxidant that prevents oxidation of low-density lipoproteins in vitro. Intake of quercetin was inversely associated with coronary heart disease mortality in elderly Dutch men. However, the extent of absorption of quercetin in humans is unclear. The aim of this study was to quantify absorption of various forms of quercetin. Nine healthy ileostomy subjects were studied, to avoid losses caused by colonic bacteria. They followed a quercetin free diet for 12 d; on days 4, 8, and 12 they received a supplement of fried onions at breakfast (rich in quercetin glucosides) equivalent to 89 mg aglycone, pure quercetin rutinoside (the major quercetin compound in tea) equivalent to 100 mg aglycone, or 100 mg pure quercetin aglycone, in random order. Subsequently, participants collected ileostomy effluent and urine for 13 h. In vitro incubations of quercetin or its glycosides with gastrointestinal fluids showed minimal degradation. Absorption of quercetin, defined as oral intake minus ileostomy excretion and corrected for 14% degradation within the ileostomy bag, was 52 +/- 15% for quercetin glucosides from onions, 17 +/- 15% for quercetin rutinoside, and 24 +/- 9% for quercetin aglycone. Mean excretion of quercetin or its conjugates in urine was 0.5% of the amount absorbed; quercetin excretion in urine was negatively correlated with excretion in ileostomy effluent (r = -0.78, n = 27). We conclude that humans absorb appreciable amounts of quercetin and that absorption is enhanced by conjugation with glucose. PMID- 7491894 TI - Clinical nutrition for MDs: reappraisal and identity. PMID- 7491893 TI - Long-chain polyunsaturated fatty acids in children with severe protein-energy malnutrition with and without human immunodeficiency virus-1 infection. AB - Fatty acids in plasma phospholipids were studied in 35 severely malnourished young children with a median age of 29 mo (range: 9-43 mo), who were either seronegative for human immunodeficiency virus-1 (HIV) (n = 16) or suffered from asymptomatic (stage P-1; n = 12) or symptomatic (stage P-2; n = 7) HIV disease. The malnourished children had significantly lower percentages (% by wt) of phospholipid arachidonic (20:4n-6, AA) and docosahexaenoic (22:6n-3, DHA) acids than 25 age-matched healthy control subjects (AA: 7.05% and 8.70% by wt; DHA: 0.92 and 2.61% by wt, P < 0.001). Body weights of malnourished children did not correlate with linoleic (18:2n-6) and alpha-linolenic (18:3n-3) acid values but were significantly and positively correlated with AA and DHA values (r = 0.40, P = 0.02 and r = 0.63, P < 0.0001, respectively). Plasma concentrations (mg/L) of total phospholipid fatty acids did not differ among seronegative, stage P-1, or stage P-2 patients. Percentage contributions of AA and eicosapentaenoic acid (20:5n-3, EPA) did not differ among those seronegative or in stages P-1 and P-2. In contrast, values of dihomo-gamma-linolenic acid (20:3n-6) were significantly (P < 0.05) lower in stage P-2 (2.38 mg/L) than in either seronegative (3.47 mg/L) or stage P-1 (3.66 mg/L) patients. We conclude that the severely malnourished children developed a depletion of both AA and DHA proportional to the degree of malnutrition.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7491895 TI - Ca2+ excitability of the ER membrane: an explanation for IP3-induced Ca2+ oscillations. AB - Recent research dealing with experiments and theoretical models of Ca2+ excitability of the endoplasmic reticulum (ER) membrane induced by inositol 1,4,5 trisphosphate (IP3) is reviewed. Ca2+ excitability refers to the ability of a small increment of cytoplasmic Ca2+ concentration ([Ca2+]i) to trigger a large [Ca2+]i pulse or oscillations. Such nonlinear regenerative behavior is conferred by the existence of IP3 channels and Ca(2+)-ATPase transporters on the ER membrane, which extends throughout the cytoplasm. Ca2+ excitability resembles the plasma membrane electrical excitability of neurons and other cells: it is driven by the ionic concentration gradient across the ER membrane (higher Ca2+ concentration inside the ER); each [Ca2+]i spike partially consumes the prestored energy that is reestablished through ATP-dependent active transport; and [Ca2+]i, the excitation variable, controls the nonlinear dynamic release rate of ER Ca2+. This review focuses on the kinetic models based on these features and on experiments dealing with the kinetic properties of [Ca2+]i-dependent gating of the IP3 receptor channel. We summarize evidence in favor of two roles for [Ca2+]i in gating the channel's opening: activation at a rapid time scale and inactivation on a slower time scale. Exploiting an analogy to the well-known Hodgkin-Huxley model for neuronal electrical excitability, we show how Ca2+ excitability of the ER membrane can be explained by these gating properties combined with the ER Ca2+ pump activity. The theory's ability to predict is illustrated by comparing calculated with experimental [Ca2+]i responses for pituitary gonadotrophs under various stimulus conditions. PMID- 7491896 TI - Magnetic fields applied to collagen-coated ferric oxide beads induce stretch activated Ca2+ flux in fibroblasts. AB - The ability to apply controlled forces to the cell membrane may enable elucidation of the mechanisms and pathways involved in signal transduction in response to applied physical stimuli. We have developed a magnetic particle electromagnet model that allows the application of controlled forces to the plasma membrane of substrate-attached fibroblasts. The system allows applied forces to be controlled by the magnitude of the magnetic field and by the surface area of cell membrane covered with collagen-coated ferric beads. Analysis by single-cell ratio fluorimetry of fura 2-loaded cells demonstrated large calcium transients (50-300 nM) in response to the magnetic force applications. Experiments using either the stretch-activated channel blocker gadolinium chloride or ethylene glycol-bis(beta-aminoethyl ether)-N,N,N',N'-tetraacetic acid to eliminate external calcium ions, or addition of extracellular manganese ions, indicated that there was a calcium influx through putative stretch-activated channels. The probability of a calcium influx in single cells was increased by higher surface bead loading and the degree of cell spreading. Depolymerization of actin filaments by cytochalasin D increased the amplitude of calcium response twofold. The regulation of calcium flux by filamentous actin content and by cell spreading indicates a possible modulatory role for the cytoskeleton in channel sensitivity. Magnetic force application to beads on single cells provides a controlled model to study mechanisms and heterogeneity in physical force stimulation of cation-permeable channels. PMID- 7491897 TI - Deoxygenation-induced alterations in sickle cell membrane cholesterol exchange. AB - Changes in a membrane sterol exchange of sickle red blood cells (SS RBC) induced by deoxygenation were studied using the fluorescent cholesterol analogue dehydroergosterol (DHE). DHE uptake by SS RBC membrane was measured by the incubation of SS RBC with small unilamellar vesicles (SUV) containing DHE. Deoxygenation of SS RBC, but not normal RBC, increased the rate of DHE uptake. DHE membrane content after 5 h of incubation with SUV in the cell-to-SUV ratio of 1:1 (mol lipid) was 16.25 +/- 0.94 and 12.22 +/- 0.85% of total sterol for deoxygenated and oxygenated cells, respectively. Membrane spicules isolated from these deoxygenated SS RBC had three-fold higher DHE content, suggesting that the increased sterol exchange was localized to spicules. When isolated spicules were incubated with DHE-SUV directly, 91 +/- 3% of membrane sterol was rapidly exchanged, in contrast to intact RBC, in which a maximum of 33% of sterol could be exchanged. The results suggest that spicule formation in SS RBC alters membrane cholesterol structure, such that a domain of cholesterol that is normally nonexchangeable becomes readily exchangeable with exogenous sterol. PMID- 7491899 TI - Ca(2+)-permeable large-conductance nonselective cation channels in rat basophilic leukemia cells. AB - Spreading of Ca2+ signals in rat basophilic leukemia (RBL) cells occurs by release of ATP. Therefore we studied the effect of ATP on membrane currents. ATP (1-10 microM) activated large-conductance channels. Single channel events were resolved in the whole cell mode. Similar channel activity was observed in RBL cells transfected with the muscarinic M1 receptor after stimulation with carbachol as well as after intracellular infusion of aluminum fluoride. Activation was independent of internal Ca2+ (0-10 microM). The channels had a conductance of 250 pS in 135 mM Na+ and 70 pS in 100 mM Ca2+. The permeability (P) ratio was PCa/PNa/PCs/PMg = 16:1:0.6:0.6. These channels may contribute to secretory responses by allowing Ca2+ entry, leading to high Ca2+ concentrations in the vicinity of the channel pore. PMID- 7491898 TI - Pharmacology of ATP-sensitive K+ currents in smooth muscle cells from rabbit mesenteric artery. AB - The inference that ATP-sensitive K+ (KATP) channels are involved in arterial responses to the synthetic K+ channel openers, hypoxia, adenosine, and calcitonin gene-related peptide, has relied on the sensitivity of these responses to the sulfonylureas glibenclamide and tolbutamide and to tetraethylammonium (TEA+). The inhibition of KATP currents by glibenclamide, tolbutamide, and TEA+ was investigated in single smooth muscle cells from rabbit mesenteric artery by use of the whole cell patch-clamp technique. The synthetic K+ channel openers pinacidil (half-activation 0.6 microM), cromakalim (half-activation 1.9 microM), and diazoxide (half-activation 37.1 microM) activated K(+)-selective currents that were blocked by glibenclamide. Elevation of pipette (intracellular) ATP concentration decreased K+ currents induced by pinacidil. Half-inhibition of KATP currents by glibenclamide and tolbutamide occurred at 101 nM and 351 microM, respectively. KATP currents were also inhibited by external TEA+, with half inhibition at 6.2 mM. The results indicate that glibenclamide is an effective inhibitor of KATP channels in arterial smooth muscle and that tolbutamide and TEA+ are much less effective. Furthermore, these results support numerous functional studies that have demonstrated that the vasorelaxations to K+ channel openers are inhibited by < 10 microM glibenclamide but not by < 1 mM TEA+. PMID- 7491900 TI - Smooth muscle contractility is modulated by myosin tail-S2-LMM hinge region interaction. AB - The functional significance of two major smooth muscle myosin isoforms, which differ in the nonenzymic COOH-terminal tail region, is not known. We report here that a 13-amino acid peptide, which mimics a region of the tail unique to the SM1 myosin isoform, inhibits contraction velocity in permeabilized smooth muscle. This peptide is shown to bind to the S2-light meromyosin (LMM) hinge region of myosin using sodium dodecyl sulfate-polyacrylamide gel electrophoresis, photoaffinity labeling, and immunoelectron microscopy. Our results suggest that novel intermolecular contacts between the tail and S2-LMM hinge regions of adjacent myosin molecules in the thick filament may modulate contractility and provide a basis for distinct isoform function. PMID- 7491902 TI - Warming macrophages to febrile range destabilizes tumor necrosis factor-alpha mRNA without inducing heat shock. AB - We have previously reported that sustained tumor necrosis factor (TNF)-alpha expression is suppressed by temperatures in the febrile range in human macrophages. In this study, we examined the mechanisms of high-temperature induced macrophage TNF suppression in the RAW 264.7 macrophage cell line. Incubating lipopolysaccharide (LPS)-stimulated RAW 264.7 cells at 40 degrees C reduced TNF secretion by 92% and peak TNF mRNA levels by 43% compared with cells incubated at 37 degrees C (P < 0.05) but did not affect levels of glyceraldehyde 3-phosphate dehydrogenase, beta-actin, or interleukin-6 mRNA. TNF mRNA half-life, measured after transcriptional arrest with actinomycin D, was reduced from 21.8 +/- 3.6 min in LPS-stimulated RAW 264.7 cells at 37 degrees C to 16.0 +/- 1.8 min at 40 degrees C (P < 0.03), but these cells at 40 degrees C did not alter transcription rate or TNF mRNA polysome association. TNF mRNA destabilization occurred at temperatures below the threshold (43 degrees C) for the generalized heat shock response in these cells. We conclude that heating macrophages to febrile-range temperatures attenuates sustained TNF expression by modulating posttranscriptional processing, including acceleration of TNF mRNA decay. PMID- 7491903 TI - Inhibition and acceleration of Na+/Ca2+/K+ exchange fluxes by Ag+ in bovine retinal rod outer segments. AB - The effect of Ag+ on Ca2+ fluxes mediated by the retinal rod Na+/Ca2+/K+ exchanger was investigated in intact bovine rod outer segments (ROS). Intracellular Na+ concentration ([Na+]in)-dependent Ca2+ influx and extracellular Na+ concentration ([Na+]out)-dependent Ca2+ efflux were monitored by changes in cytosolic free Ca2+ measured with the fluorescent Ca(2+)-indicating dye fluo 3. Ag+ was the most effective inhibitor of Na+/Ca2+/K+ exchange fluxes described to date, with half-maximal inhibition observed at 2-8 microM Ag+. Inhibition by Ag+ could be reversed by addition of beta-mercaptoethanol but not by addition of cysteine. Reversal by beta-mercaptoethanol resulted in a marked acceleration of [Na+]out-dependent lowering of cytosolic free Ca2+ but not of [Na+]in-dependent Ca2+ influx. We suggest that Ag+ inhibits and accelerates Na+/Ca2+/K+ exchange fluxes by binding to cysteine residues on the cytosolic surface of the exchanger protein. PMID- 7491901 TI - The mitogenic effect of platelet-derived growth factor in human hepatic stellate cells requires calcium influx. AB - Platelet-derived growth factor (PDGF) is a key mitogen for hepatic stellate cells (HSC) and has been shown to be implicated in liver tissue repair and fibrogenesis. In this study the relationship between PDGF-induced intracellular Ca2+ concentration ([Ca2+]i) increase and mitogenesis in cultured human HSC was evaluated. In high-density cell cultures (80-90% subconfluence), PDGF induced a significant increase in [Ca2+]i, characterized by a short-lasting peak phase, which was followed by a long-lasting plateau phase. The plateau phase was abolished in the absence of extracellular Ca2+. However, in low-density cell cultures (30-40% subconfluence), the plateau phase was absent or markedly less pronounced. In parallel sets of experiments, PDGF was significantly less effective in inducing mitogenesis in low-density cell cultures than in high density cell cultures and was totally ineffective in the absence of extracellular Ca2+. These results suggest that 1) spatial and time dynamics of PDGF-induced [Ca2+]i increase are dependent on cell density and 2) PDGF-induced mitogenesis requires extracellular Ca2+ influx. PMID- 7491904 TI - Ca2+ influx into bovine retinal rod outer segments mediated by Na+/Ca2+/K+ exchange. AB - Ca(2+)-depleted rod outer segments (ROS) were purified from bovine retinal rod photoreceptors, and factors influencing Ca2+ influx into ROS via the plasma membrane Na+/Ca2+/K+ exchanger were analyzed. Intracellular alkali cation concentrations were manipulated by 1) previous loading via the ionophore monensin followed by removal of monensin and 2) addition of the channel ionophore gramicidin during Ca(2+)-influx measurements. Ca2+ influx was measured as a rise in cytosolic free Ca2+ with the Ca(2+)-indicating dye fluo 3. An absolute requirement for intracellular Na+ was observed with a Na+ dissociation constant of 30-40 mM, whereas intracellular K+ was a potent inhibitor of Ca2+ influx, apparently by competing with Na+ for a common site on the Na+/Ca2+/K+ exchanger. Half-maximal Ca2+ influx was observed at an external free Ca2+ concentration of 0.9 microM when no external Na+ was present and 3.5 microM when 10 mM external Na+ was present. Our observations are discussed in the context of 1) a three-site model for the Na+/Ca2+/K+ exchanger and 2) earlier experiments on light adaptation in rods, which depended on minimizing Ca2+ fluxes across the ROS plasma membrane. PMID- 7491905 TI - Thiophosphorylation independently activates each head of smooth muscle myosin in vitro. AB - To determine whether thiophosphorylation of the 20-kDa myosin light chain activates each head of smooth muscle myosin independently of the head with which it is paired, chicken gizzard smooth muscle myosin was randomly thiophosphorylated, producing a mixture of unphosphorylated and singly and doubly thiophosphorylated myosin. Thiophosphorylation levels were measured by glycerol urea gels, and the activity of this myosin was determined by actin-activated adenosinetriphosphatase measurements and in an in vitro motility assay, where the velocity of actin filaments moving over a myosin-coated surface is measured. Activity at each thiophosphorylation level was similar to that previously observed for mixtures of unphosphorylated and doubly thiophosphorylated myosin (D. E. Harris, S. S. Work, R. K. Wright, N. R. Alpert, and D. M. Warshaw. J. Muscle Res. Cell Motil. 15: 11-19, 1994). All doubly thiophosphorylated myosin was then formed into filaments and removed from randomly thiophosphorylated myosin by centrifugation. The remaining myosin (mixture of unphosphorylated and singly phosphorylated myosin), which could not polymerize because of their conformation, retained approximately 70% activity compared with mixtures of unphosphorylated and doubly thiophosphorylated myosin. Thus a thiophosphorylated smooth muscle myosin head can produce substantial biochemical and mechanical activity, even when it is paired with an unphosphorylated partner. PMID- 7491906 TI - Temperature-induced functional deocclusion of thiols inhibitory for sheep erythrocyte K-Cl cotransport. AB - In low-K sheep erythrocytes, K-Cl cotransport is activated by treatment with low concentrations of thiol reagents and by other interventions such as lowering of cellular free cytosolic Mg, hyposmotic cell swelling, the kinase inhibitor staurosporine, and hydroxylamine. High concentrations of N-ethylmaleimide or methylmethane thiolsulfonate reverse the activation through thiol groups and, as shown here, also the stimulation by non-thiol manipulations. The overriding inhibitory sites functionally associated with and different from those of the activating thiols (SHa) were further distinguished by temperature. Treatment with N-ethylmaleimide and its subsequent removal by dithiothreitol, both at 0 degrees C, prevented the inhibitory effect at 37 degrees C and thus the chemical modification of inhibitory thiols (SHi). Whereas stimulation through SHa closely followed the loss of glutathione, inhibition through SHi occurred only in glutathione-depleted cells. The reversal of K-Cl cotransport stimulation by all hitherto known interventions, which is strongest in metabolically depleted cells, suggests that the low temperature-protected SHi constitute crucial sites that, close to the transporter itself or at the cytoskeletal level, become functionally deoccluded upon temperature elevation. PMID- 7491907 TI - Endothall thioanhydride inhibits protein phosphatases-1 and -2A in vivo. AB - The objective of this study was to relate the toxicity of several cantharidin derivative pesticides with their abilities to inhibit protein phosphatases-1 (PP1) and -2A (PP2A). Cantharidin (CA), endothall, and endothall thioanhydride (ETA) inhibited the activity of PP1 and PP2A, and the potency sequence was CA > endothall > ETA in vitro. We determined the inhibitory potency of these pesticides on hepatic protein phosphatases by administration of the toxins into the portal vein of rats. The potency sequence of ETA > CA > endothall was established for the inhibition of PP1 and PP2A in vivo and shows close correlation with the sequence of relative toxicity. ETA predominantly targets PP1 for inhibition in liver, as revealed by assays specific for PP1 or PP2A. Studies using 3T3 fibroblasts showed that only ETA, but not CA or endothall, induced marked morphological changes. These effects included cell rounding and detachment as well as extensive reorganization of actin filaments and are characteristic for the cell-permeable phosphatase-inhibitory toxins. It is suggested that the in vivo effectiveness is related to enhanced uptake of ETA, because this is permeable across the plasmalemma. PMID- 7491908 TI - Suppression of a nonselective cation conductance by substance P in cochlear outer hair cells. AB - Substance P (SP) hyperpolarizes outer hair cells (OHCs) of guinea pig cochlea. The cellular mechanisms of the SP response were investigated with the whole cell patch-clamp technique. SP induced outward currents in a dose-dependent manner at a holding potential of -60 mV in a concentration range between 3 x 10(-6) and 10( 4) M. SP decreased slope conductance between -60 and +20 mV. Ion substitutions in the external medium revealed that SP suppresses nonselective cation conductance with high permeability for Ca2+. The relative ion permeability of the channel modulated by SP was as follows: Ca2+ > Li+ approximately Cs+ approximately Na+ > Tris+. The potency of the agonist action was as follows: SP >> neurokinin A > neurokinin B. Peptide antagonists induced currents similar to those of SP. CP 96345, a selective nonpeptide antagonist for the neurokinin type 1 receptor, did not inhibit the SP-induced current. Intracellular dialysis of guanosine 5'-O-(2 thiodiphosphate) and pertussis toxin (PTX) suggests that a PTX-insensitive G protein is involved in the SP response. Neither the Ca2+ chelator 1,2-bis(2 aminophenoxy)ethane-N,N,N',N'-tetraacetic acid nor staurosporine (10(-6) M) affects the SP response. Local application of SP by a puffer pipette indicates that the SP receptors are distributed along the side of the OHC. These results suggest the possibility that the action of SP on the OHCs may not be mediated by the tachykinin receptors but rather by a tachykinin receptor-independent pathway. It is proposed that SP suppresses the nonselective cation conductance in the lateral wall of OHCs via a PTX-insensitive G protein. PMID- 7491909 TI - Effects on Ca(2+)-activated tension due to a synthetic NH2-terminal actin peptide in single skeletal muscle fibers. AB - Insight into the mechanism of force development in striated muscle will be provided by elucidating the specific regions of the actin molecule that interact with myosin and regulatory subunits of the thin filament during Ca(2+)-activated contraction. There is growing evidence that the acidic NH2-terminal domain of actin 1) may represent an important binding site for myosin and 2) may interact with the inhibitory region of troponin I. The purpose of this study was to determine the effects of a synthetic peptide corresponding to a specific sequence of the NH2-terminal domain of skeletal muscle actin on Ca(2+)-activated tension in chemically skinned single psoas skeletal muscle fibers. This study focused on the highly conserved Lys18-Arg28 amino acid sequence of actin, a region of native actin that is believed to interact with troponin I and myosin. The effects of synthetic actin peptide Lys18-Arg28 on tension development varied, depending on 1) the concentration of Ca2+ in the activating solutions and 2) the peptide concentration. At submaximal concentrations of Ca2+, isometric tension was reversibly potentiated in the presence of 100-500 microM synthetic actin peptide Lys18-Arg28. Importantly, scrambling the sequence of Lys18-Arg28 fully abolished the increase in Ca2+ sensitivity, providing evidence that the observed effects were specific to the sequence of peptide Lys18-Arg28. In contrast, maximum Ca(2+) activated tension was inhibited by millimolar concentrations of Lys18-Arg28 and the scrambled peptide, indicating that this effect was nonspecific. The effect of peptide Lys18-Arg28 to increase the Ca2+ sensitivity of tension is not known but may be due to an effect of the actin peptide to alter thin filament activation, a possibility consistent with proposed interactions between this domain of actin and the inhibitory region of troponin I. PMID- 7491910 TI - Activation of a selective pathway of lysosomal proteolysis in rat liver by prolonged starvation. AB - Lysosomal uptake and degradation of polypeptides such as glyceraldehyde-3 phosphate dehydrogenase (GAPDH), ribonuclease A (RNase A), and RNase S-peptide (residues 1-20 of RNase A) are progressively activated in rat liver by starvation before isolation of lysosomes. This pathway of proteolysis is selective, since it is stimulated by the heat shock cognate protein of 73 kDa (HSC73) and ATP-MgCl2, and lysosomal uptake of RNase A could be competed by GAPDH but not by ovalbumin. A portion of intracellular HSC73 is associated with certain lysosomes, and the amount of lysosomal HSC73 increases by 5- to 10-fold during prolonged starvation. The lysosome-associated HSC73 is primarily within the lysosomal lumen. Double immunogold labeling of lysosomes incubated in vitro with RNase A detects this protein substrate as well as HSC73 within lysosomes. More than two-thirds of the labeled lysosomes contain both RNase A and HSC73. The possible physiological significance of the activation of this selective pathway of lysosomal proteolysis in long-term starvation is discussed. PMID- 7491911 TI - Micro- and macrovascular endothelial cells in beta-adrenergic regulation of transendothelial permeability. AB - Barrier function of endothelial cells (EC) was modulated using beta-adrenergic agonists, e.g., isoproterenol (ISO) and formoterol (FOR). To get a direct comparison between EC from different vascular sources, we isolated EC from aorta (BAEC) and retina (BREC) of the same calf. For permeability studies, EC were cultured on polycarbonate filters. At confluency, transendothelial exchange of the diffusion marker fluorescein isothiocyanate-dextran was determined. Microvascular retinal EC monolayers are half as permeable as monolayers from macrovascular BAEC. When EC are stimulated with beta-adrenergic receptor (AR) agonists, monolayer permeability decreases, and the amount of intracellular adenosine 3',5'-cyclic monophosphate (cAMP) increases in both cell types. Comparison of the half-maximum concentrations causing change in permeability (pEC50) shows direct coupling between beta-AR and adenylate cyclase. The beta 2 selective agonist FOR stimulates cAMP synthesis in BAEC with a pEC50 value of 9.37 and decreases permeability with a pEC50 value of 9.72. In BREC, the pEC50 values of ISO concerning stimulation of cAMP synthesis and the decrease of permeability are also very similar, 5.32 and 5.34, respectively. BREC are not as sensitive to beta 2-AR agonists as BAEC. The pEC50 value of FOR influence on BREC permeability is 8.77 in comparison with 9.72 for BAEC. These results could be interpreted with different affinities of the beta 2-selective AR agonist in BREC and BAEC. PMID- 7491912 TI - Intracellular Ca2+ stores in chick cerebellum Purkinje neurons: ontogenetic and functional studies. AB - The molecular composition of intracellular Ca2+ stores in developing chicken cerebellum Purkinje neurons from embryonic day 11 (E11) to posthatching day 2 (P2) was studied by immunocytochemistry using specific antibodies for three molecular constituents, the receptor (R) and/or channel sensitive to inositol 1,4,5-trisphosphate (IP3), Ca(2+)-adenosinetriphosphatase (ATPase), and calsequestrin (CS). CS, IP3R, and Ca(2+)-ATPase were first detected by light microscopic immunofluorescence in migrating Purkinje cells at E11-E12 and throughout late phases of embryonic development. Ontogenesis of CS, IP3R, and Ca(2+)-ATPase accompanied well-defined stages of cerebellum histogenesis and cytogenesis and was accomplished before hatching. High-resolution immunogold electronmicroscopy revealed that, at E18-P1, CS was still largely distributed to the endoplasmic reticulum (ER) lumen and began to be segregated to ER subcompartments (calciosomes) only by P2, whereas the IP3R was concentrated into ER cisternal stacks as early as E18. Both ionotropic and metabotropic plasma membrane receptors were present in dissociated single chicken Purkinje cells from E16 onward, as indicated by measurements of membrane currents (whole cell recording mode) and of cytoplasmic Ca2+ transients monitored with the cell trappable fluorescent indicator fura 2-acetoxymethyl ester, respectively. Cytoplasmic Ca2+ transients were detected after either activation of glutamate metabotropic receptors, i.e., evidence of IP3-sensitive Ca2+ channels, or application of caffeine, i.e., evidence of ryanodine-sensitive Ca2+ channels. Intracellular Ca2+ stores appear to be functional during embryonic development. PMID- 7491913 TI - Stimulation of GLUT-1 glucose transporter expression in response to exposure to calcium ionophore A-23187. AB - We tested the hypothesis that an increase in cytosolic calcium concentration stimulates glucose transporter isoform (GLUT-1) gene expression. Exposure of a rat liver cell line (Clone 9) to 3 microM A-23187 for 12 h resulted in 3-, 5-, and 10-fold increases in cytochalasin B-inhibitable 3-O-methyl-D-glucose transport, GLUT-1 protein, and GLUT-1 mRNA content, respectively. The induction of GLUT-1 mRNA in response to A-23187 is not preceded by a significant decrease in cell ATP content. This induction is prevented by 1,2-bis(2-aminophenoxy)ethane N,N,N',N'-tetraacetic acid in conjunction with ethylene glycol-bis(beta aminoethyl ether)-N,N, N',N'-tetraacetic acid. To investigate the mechanism of GLUT-1 mRNA induction, we found that exposure to A-23187 stabilized GLUT-1 mRNA: with the employment of actinomycin D, GLUT-1 mRNA had a half-life of 1.5 and 5.5 h in control and A-23187-treated cells, respectively. In nuclear run-on assays, the rate of GLUT-1 gene transcription was stimulated 1.5- to 1.7-fold in nuclei isolated from cells exposed to A-23187 for either 30 min or 2 h. These results demonstrate that exposure to A-23187 stimulates GLUT-1 gene expression and that the increase in GLUT-1 mRNA content is mediated in part by enhanced GLUT-1 gene transcription as well as decreased GLUT-1 mRNA degradation. The increase in GLUT 1 mRNA content, in turn, is associated with increased cell GLUT-1 content and enhanced glucose transport. PMID- 7491914 TI - Ion channel classes in purified olfactory cilia membranes: planar lipid bilayer studies. AB - Ciliary membrane fragment fusion to planar lipid bilayers resulted in the insertion of four ion channel types. cAMP-activated, cation-selective channels could be detected only in the absence of Ca2+ and had a conductance of 23 pS. They exhibited an apparent dissociation constant (Kd) for the cyclic nucleotide of approximately 30 microM and an estimated permeability ratio (PNa/PK) of 2.4. The cAMP cation-selective channel coinserted with a K(+)-selective channel refractory to cAMP, Ca2+, and D-myo-inositol 1,4,5-trisphosphate. This K+ channel was voltage independent and exhibited open-conductance substates of 60 and 112 pS. cAMP was also found to modulate a novel K+ channel with a Kd = 140 microM. It displayed three nearly equally spaced open substates with conductances of 34, 80, and 130 pS. In the absence and in the presence of cAMP the probability of occurrence of the open substates was binomially distributed. A fourth channel type was a Ca(2+)-activated K+ channel with a conductance of 240 pS. It was blocked by charybdotoxin at nanomolar concentrations (Kd = 3 nM). These results add support to the idea that, besides cAMP-activated cation-selective channels, vertebrate chemosensory olfactory membranes possess an arrangement of ion channels. PMID- 7491915 TI - Anemia in copper-deficient rats: role of alterations in erythrocyte membrane fluidity and oxidative damage. AB - This study was designed to make precise the nature and the mechanism of the anemia induced by dietary copper (Cu) deficiency. Male Wistar rats were pair fed from weanling for 6 wk either a Cu-deficient or a control diet. The reduced red blood cell (RBC) 51Cr survival indicates an increased destruction of RBC during Cu deficiency. 1,6-Diphenyl-1,3,5-hexatriene fluorescence polarization studies revealed an increase in the fluidity of erythrocyte membranes from deficient rats. The reduced cholesterol-to-phospholipid ratio was consistent with the increased fluidity. Other results indicate an increased vulnerability of RBC to hemolysis in dilute hydrogen peroxide and an increased formation of lipid peroxidation products. Before exposure to free radical stress, electron spin resonance studies in intact RBC revealed decreased correlation time of 16-doxyl stearic acid, confirming a more fluid membrane in RBC from Cu-deficient rats. After in vitro peroxidation, RBC from Cu-deficient rats showed a more ordered state of membrane lipids compared with controls. Together, these studies demonstrate the hemolytic nature of the anemia. The shortened survival of erythrocytes apparently results from changes in membrane fluidity and enhanced susceptibility to peroxidation. PMID- 7491916 TI - Dynamics of pancreatic cell growth and differentiation during diabetes reversion in STZ-treated newborn rats. AB - Cellular processes underlying ontogenesis and regression of streptozotocin (STZ) induced diabetes in newborn rats were investigated at the most severe stage of diabetes at day 3 and after recovery of normoglycemia at day 8 by immunocytochemistry and quantitative analysis. A previously unknown endocrine cell type subpopulation (PEPS) was identified. It was characterized by granule polymorphism, coexpression of insulin and glucagon immunoreactivity, and a proliferative capacity transiently higher than in B cells. In STZ-treated rats at day 3, B cell mass decreased 14-fold, whereas PEPS cells were unaffected. The islet mass was restored to 55.7% by day 8, with a concomitant appearance of numerous small islets contiguous to small ducts. B cell mass increased by 6.9 fold compared with 1.8-fold in control rats, although proliferative capacities remained similar. Proliferation dropped considerably by day 8, preventing complete B cell mass recovery in STZ-treated rats. STZ-induced neonatal diabetes thus stimulates neogenesis of islets close to ducts and proliferation of PEPS cells. Those partially differentiated islet cells appear to be on the differentiation pathway of stem cells to fully differentiated B cells. PMID- 7491917 TI - Nuclear responses to depletion of mitochondrial DNA in human cells. AB - The derivation of human cell lines devoid of mitochondrial (mt) DNA (rho 0) provides an opportunity to study nuclear responses to a chronic impairment of mitochondrial oxidative phosphorylation. Expression of several nuclear genes is induced in human rho 0 cells, including those encoding integral proteins of the mitochondrial inner membrane, intermediate filaments, and ribosomes. In contrast to conditions in which mitochondrial respiration is altered acutely, expression of heat shock proteins and immediate early genes is not induced. Mitochondria from rho 0 cells maintain a transmembrane electrochemical potential and are distributed within the cytoplasm of these cells in a manner indistinguishable from that of wild-type cells. We conclude that a chronic deficiency of mitochondrial oxidative phosphorylation produced by elimination of mtDNA is associated with a different pattern of gene induction than that provoked by other acute or subacute conditions that impair mitochondrial respiration or create energy demands in excess of mitochondrial respiratory capacity. PMID- 7491918 TI - Effects of fluoride and cholera and pertussis toxins on sensory transduction in the carotid body. AB - The regulation of the chemoreceptor cell function by G proteins has been studied by measuring the release of 3H-labeled catecholamines ([3H]CA) in carotid bodies (CBs) treated with fluoride, cholera toxin (CTX), and pertussis toxin (PTX). Fluoride augmented the basal release of [3H]CA in a dose- (5-20 mM) and Ca(2+) dependent manner. Nisoldipine (1 microM) and ethylisopropyl amiloride (EIPA; 10 microM) inhibited this effect by approximately 60%, and both drugs combined inhibited it in full. BAY K 8644 (1 microM) doubled the effect of fluoride. The effects of fluoride on the stimulus-evoked release of [3H]CA varied with the type of stimulus and the duration of the treatment. Simultaneous application of fluoride with the stimulus increased by five times the release evoked by hypoxia and by two times that by K+ and dinitrophenol (DNP). Preincubation with fluoride for 1 h caused an inhibition (approximately 70%) of the release evoked by high K+ and veratridine, whereas that evoked by DNP and low PO2 was still augmented (approximately 2 times). Preincubation (4 h) of the CBs with CTX (3 micrograms/ml) reduced by 54% the release of [3H]CA evoked by 35 mM K+ but did not affect that evoked by low PO2 or DNP. A similar treatment with PTX (1 microgram/ml) affected only the release of [3H]CA evoked by DNP, reducing it by 65%. The data show that fluoride, CTX, and PTX have different effects on the release of [3H]CA evoked by high external K+, DNP, and low PO2, indicating that the stimulus-secretion coupling process for each stimulus is differently regulated by G proteins. PMID- 7491919 TI - Volume-associated osmolyte fluxes in cell lines with or without the anion exchanger. AB - To investigate the involvement of a red cell-type anion exchanger in the volume sensitive amino acid release, the hyposmolarity-evoked release of D-[3H]aspartate and [3H]taurine was examined in three cell lines: 1) wild-type Chinese hamster ovary (CHO-K1) cells, expressing an anion exchanger activity (Cl-/SO4(2-)) functionally similar to the erythroid band 3; 2) a mutant CHO cell type (CHO 605) lacking this anion exchanger activity; and 3) 293 cells in which the Cl-/HCO3(-) anion exchanger is absent. All cell types accumulated D-[3H]aspartate and [3H]taurine under isosmotic conditions, and, similarly, in the three cell lines, cell swelling evoked by hyposmolarity induced a rapid and transient increase in the amino acid efflux. Blockers of the anion exchanger and/or Cl- channels [niflumic acid, dipyridamole, diphenylamine-2-carboxylate,5-nitro-2-(3 phenylpropylamino)-benzoi c acid, and 4,4'-diisothiocyanostilbene-2,2'-disulfonic acid] were potent inhibitors of amino acid efflux in the three cell lines. 125I- efflux, used as a marker for Cl- fluxes, was also markedly increased in response to cell swelling in all cell lines, and this efflux was inhibited by the anion exchanger/Cl- channel blockers. These results do not support a role for an anion exchanger in the hyposmolarity-induced amino acid efflux and suggest that amino acids and Cl- may be transported by the same or a similar mechanism, presumably an anion channel-like structure. PMID- 7491920 TI - Leucine activates system A amino acid transport in L6 rat skeletal muscle cells. AB - In this study, we present evidence showing that leucine is involved in the upregulation of system A amino acid transport activity in the L6 rat skeletal muscle cell line. At leucine concentrations of > or = 0.05 mM, the uptake of N methylamino-alpha-isobutyric acid (MeAIB), a paradigm system A substrate, was stimulated by up to 50%. Kinetic analysis revealed that this stimulation was a result of an increase in the maximal transport rate of MeAIB uptake, from 327 +/- 26 to 450 +/- 8 pmol.min-1.mg protein-1 after incubation of cells with leucine. No significant change in the concentration at which MeAIB transport was half maximal was observed. System A activation was biphasic, reaching an initial plateau after 3 h, with a second phase of activation being observed after 5 h. The initial activation of system A transport occurred by a mechanism distinct from that activated by insulin-like growth factor-I (IGF-I) (3 nM), since the effects of leucine and IGF-I were additive. This activation was not due to transstimulation, since 2-amino-2-norbornane-carboxylic acid, a specific system L substrate, did not stimulate system A. Leucine's keto acid, ketoisocaproic acid, prevented the activation of system A transport, whereas aminooxyacetate, a transaminase inhibitor, augmented the increase in system A activity by leucine. Both cycloheximide and actinomycin D inhibited the leucine-induced increase in MeAIB uptake. The present results indicate that leucine, or some cellular component regulated by it, is capable of stimulating system A transport through control of DNA transcription, possibly of a gene encoding either a repressor or enhancer molecule of system A or perhaps of the gene encoding system A itself. PMID- 7491921 TI - Relationship between severity, necrosis, and apoptosis in five models of experimental acute pancreatitis. AB - In an effort to elucidate factors that determine the severity of an attack of acute pancreatitis, we have quantitated the extent of necrosis and of apoptosis in five different models of experimental acute pancreatitis. Severe pancreatitis was induced by obstructing the opossum common bile-pancreatic duct, by administering to mice 12 hourly injections of a supramaximally stimulating dose of caerulein, and by feeding young female mice a choline-deficient, ethionine supplemented diet. In each of these models of severe pancreatitis, marked necrosis but very little apoptosis was found. Mild pancreatitis was induced by obstructing the rat common bile-pancreatic duct and by infusing rats with a supramaximally stimulating dose of caerulein. In contrast to our findings in severe pancreatitis, mild pancreatitis was characterized by very little necrosis but a high degree of apoptosis. Our finding that the severity of acute pancreatitis is inversely related to the degree of apoptosis suggests that apoptosis may be a teleologically beneficial response to acinar cell injury in general and especially in acute pancreatitis. PMID- 7491923 TI - Induction of hypertrophic responsiveness to isoproterenol by TGF-beta in adult rat cardiomyocytes. AB - In a previous publication we reported that hypertrophic responsiveness to beta adrenoceptor stimulation can be induced in isolated cardiomyocytes when these are cultured for 6 days in presence of fetal calf serum (FCS; Pinson et al., J. Mol. Cell. Cardiol.. 25: 477-490, 1993). The role of transforming growth factor-beta (TGF-beta) in this induction process has now been investigated. Isolated cardiomyocytes from adult rats were cultured for 6 days in presence of 20% FCS. It was found that induction of hypertrophic responsiveness to beta-adrenoceptor stimulation was abolished when a neutralizing anti-TGF-beta 1 antibody was added to FCS-containing culture medium. In culture media with FCS contents (5%) too low to induce hypertrophic responsiveness to beta-adrenoceptor stimulation, addition of 1 ng/ml TGF-beta 1,2 induces this responsiveness. It was demonstrated that cardiomyocytes already release TGF-beta into culture media on day 1 of culture and that they continue to do so in presence of FCS supplements of > 5%. The results demonstrate that hypertrophic responsiveness to beta-adrenoceptor stimulation is induced in cardiomyocytes by an autocrine mechanism involving TGF beta 1 as mediator. PMID- 7491922 TI - Dexamethasone enhances expression of mitochondrial oxidative phosphorylation genes in rat distal colon. AB - Dexamethasone and aldosterone are major activators of Na+ reabsorption in tight epithelia. The genes whose expression mediates the steroid actions are mostly unknown. To identify such genes, we performed differential screening of a rat colon cDNA library with total 32P-labeled cDNA probes reverse transcribed from steroid-stimulated and steroid-depleted poly(A)+ RNA. Several cDNAs whose corresponding mRNA is enhanced two- to threefold after dexamethasone injection were identified. Partial sequencing indicated that four of them code for subunits of cytochrome-c oxidase and 16S mitochondrial mRNA. The dexamethasone-induced increase in mitochondrial RNA abundance could not be mimicked by a low-salt diet, found to increase plasma aldosterone from 1.0 +/- 0.1 to 12.8 +/- 1.4 nM. Induction of mitochondrial genes by adrenal steroids may serve to prevent limitation of transport by the ATP supply to the Na(+)-K+ pump under conditions of maximal stimulation of Na+ transport. PMID- 7491924 TI - Integrative effects of EGF on metabolism and proliferation in renal proximal tubular cells. AB - This study examined the relationship between alterations in cellular metabolism and induction of proliferation in renal proximal tubular cells (RPTC) after epidermal growth factor (EGF) exposure. EGF treatment (10 ng/ml) of confluent RPTC cultures for 6 consecutive days increased monolayer DNA content 3.3-fold. EGF-stimulated proliferation of RPTC was preceded by a rapid (within 4 h) induction of glycolysis and a decrease in basal and ouabain-sensitive oxygen consumption (20 and 30%, respectively). EGF stimulated the pentose cycle by 58% and decreased gluconeogenesis by 48%. Supplementation of the culture medium with ribose-5-phosphate or ribose abolished the stimulation of glycolysis and the pentose cycle by EGF but had no effect on proliferation. These results show that EGF rapidly stimulates the pentose cycle, shifts glucose metabolism from gluconeogenesis to glycolysis, and decreases oxygen consumption before any increase in proliferation. The lack of an EGF effect on the pentose cycle and glycolysis in the presence of exogenous precursors for DNA synthesis suggests that the stimulation of these pathways before proliferation is due to increased demands for ribose for subsequent nucleic acid synthesis. PMID- 7491925 TI - Prolactin inhibits corticoid-induced differentiation of active Na+ transport across cultured frog tadpole skin. AB - Active Na+ transport differentiates in larval bullfrog skin cultured with corticoids. After 2 wk in culture, the epidermis became positive against human blood group antigen A, the marker for the adult-type cells of the epidermis, but was negative to the antibody against the acetylcholine receptor, the marker for the larval-type epidermis. Amiloride (10(-5) M) did not inhibit the differentiation of active Na+ transport. On the other hand, in skin cultured with prolactin (2 micrograms/ml), the epidermis remained negative against antigen A and positive against acetylcholine receptor, and the differentiation of active Na+ transport was inhibited. Thyroid hormone did not antagonize the inhibitory action of prolactin on this transport differentiation. Prolactin affected the basal cells of the larval epidermis and inhibited development of corticoid induced adult features in the epidermis. PMID- 7491926 TI - Ins(1,4,5)P3 activates Drosophila cation channel Trpl in recombinant baculovirus infected Sf9 insect cells. AB - The trp-like (trpl) gene product (Trpl) is thought to form a nonselective cation channel important for signal transduction in Drosophila photoreceptor cells. This channel may be the insect homologue of mammalian channels involved in Ca2+ signal transduction. To determine the mechanism of receptor-mediated activation of Trpl, whole cell membrane currents were examined in Sf9 insect cells after infection with recombinant baculovirus. Stimulation by bradykinin increased whole cell Trpl currents three- to fivefold. Similar activation of Trpl was observed by inclusion of D-myo-inositol 1,4,5-trisphosphate [Ins(1,4,5)P3] in the pipette solution during whole cell recordings. These currents were 1) not seen in noninfected cells or in cells expressing only the B2 receptor, 2) mimicked by D-myo-inositol 2,4,5-trisphosphate, and 3-deoxy-3-fluoro-D-myo-inositol 1,4,5-trisphosphate, 3) not seen with D-myo-inositol 1,4-bisphosphate or D-myo-inositol 1,3,4,5 tetrakisphosphate, and 4) blocked by heparin, but not by de-N-sulfated heparin. In contrast, Trpl currents were unaffected by thapsigargin. These results demonstrate that the Trpl cation channel is activated by Ins(1,4,5)P3 in a heparin-sensitive fashion. Regulation of channel activity by Ins(1,4,5)P3 may occur by a number of mechanisms, including direct binding of Ins(1,4,5)P3 to the Trpl channel or direct physical interaction between the Ins(1,4,5)P3 receptor/Ca(2+)-release channel of the endoplasmic reticulum and the Trpl protein. PMID- 7491927 TI - Antisense inhibition of Na+/Ca2+ exchange in primary cultured arterial myocytes. AB - The effects of chimeric phosphorothioated antisense oligodeoxynucleotides (AS oligos) targeted against the Na+/Ca2+ exchanger (NCX) were tested in primary cultured rat mesenteric artery myocytes. In parallel cultures, myocytes proliferated and were morphologically normal in the presence of scrambled nonsense (NS-) or AS-oligos or no oligos (controls). NCX function was examined with digital imaging, using fura 2 to estimate the cytosolic free Ca2+ concentration ([Ca2+]cyt). Resting [Ca2+]cyt was higher (145 +/- 4 nM; P < 0.05) in AS-oligo-treated cells than in controls (125 +/- 5 nM) or NS-oligo-treated cells (131 +/- 4 nM). Lowering external Na+, to promote Ca2+ entry via NCX, increased [Ca2+]cyt transiently in controls and NS-oligo-treated cells but not in AS-oligo-treated cells. Raising the cytosolic free Na+ concentration with ouabain augmented the low-Na(+)-induced rise in [Ca2+]cyt in controls and NS-oligo treated cells, but AS-oligo-treated cells still did not respond. Nevertheless, serotonin (5-HT) increased [Ca2+]cyt in all three groups. Thus AS-oligos selectively blocked NCX activity but not the 5-HT response. To determine the effect of NCX knockdown on the modulation of stored Ca2+, the 5-HT response was tested immediately after removal of external Ca2+. Ouabain augmented the 5-HT induced rise in [Ca2+]cyt in control and NS-oligo-treated cells but not AS-oligo treated cells. This indicates that the NCX can modulate intracellular Ca2+ stores. We conclude that AS-oligos are useful for investigating the physiological role of NCX in vascular smooth muscle. PMID- 7491928 TI - Transgenic animals in the study of blood pressure regulation and hypertension. AB - It is generally accepted that the etiology of essential hypertension is due to a complex interplay of genetic and environmental factors. A great deal of research effort over the past ten years has been focused on the identification of genes the variants of which predispose individuals to high blood pressure. Consequently, transgenic and knockout animals have become important research tools, providing experimental systems in which defined genetic manipulations can be introduced on uniform genetic backgrounds while minimizing environmental variation. These animal models have provided the means by which candidate genes thought to be involved in blood pressure regulation have been studied. Furthermore, these models can be used to test the significance of genes and gene variants identified via genome-wide searches as potential causes of hypertension. The purpose of this review is to provide a brief discussion of transgenic and knockout methodology and its application to study the genetic basis of hypertension. PMID- 7491929 TI - Expression of IGF-II and IGF binding proteins in differentiating human intestinal Caco-2 cells. AB - The mitogenic and metabolic effects of insulin-like growth factor-II (IGF-II) can be modulated by six distinct IGF binding proteins (IGFBPs). As a first step toward understanding the role of IGFs and their binding proteins in intestinal epithelial cell differentiation, the expression of IGF-II and IGFBPs was characterized in the human colon adenocarcinoma Caco-2 cell line. Northern blot analysis revealed two IGF-II transcripts of 5.4 and 4.5 kb, and ribonuclease protection assays indicated that IGF-II mRNA levels are regulated during Caco-2 differentiation. A specific radioimmunoassay detected IGF-II in serum-free conditioned medium, the level of which was three- to fivefold higher in proliferating cells than in differentiated cells. Immunoprecipitation and ligand blot analyses of conditioned medium demonstrated that IGFBP-2, IGFBP-3, IGFBP-4, and IGFBP-6 are synthesized by Caco-2 cells, with IGFBP-2 and IGFBP-4 being the major IGFBPs secreted, and that the levels of IGFBP-2 and IGFBP-6 decreased as differentiation proceeded. These results indicate that the expression of IGF-II, IGFBP-2, and IGFBP-6 is regulated in a differentiation-dependent manner in Caco-2 cells. PMID- 7491930 TI - Individual lactotropes release prolactin in a temporally divergent and sexually dimorphic pattern. AB - Recent studies reveal that individual lactotropes vary in the amount of prolactin (PRL) released from day to day and that the magnitude of these variations is gender specific. In an attempt to further elucidate the dynamics of hormone secretion by single cells, we have investigated short-term variations in PRL release from male and lactating female rat lactotropes. To this end, hormone release from individual lactotropes was monitored at 12-min intervals for up to 2 h, using a modified version of the reverse hemolytic plaque assay. We found that pituitary cells from both genders could release PRL in either an intermittent or continuous mode, but that the former pattern of secretion predominated for male lactotropes. Interestingly, the mode of hormone secretion was tightly coupled (P < 0.001) to the amount of hormone released (continuous >> intermittent), regardless of the gender of the pituitary donor. The existence of gender-specific differences in secretory modalities indicates that this fluctuating pattern of hormone secretion by individual cells is not random but regulated, perhaps through entrainment in vivo by gonadal steroids. PMID- 7491931 TI - Testosterone administration to elderly men increases skeletal muscle strength and protein synthesis. AB - Aging men develop a significant loss of muscle strength that occurs in conjunction with a decline in serum testosterone concentrations. We investigated the effects of testosterone administration to six healthy men [67 +/- 2 (SE) yr] on skeletal muscle protein synthesis, strength, and the intramuscular insulin like growth factor I (IGF-I) system. Elderly men with serum testosterone concentrations of 480 ng/dl or less were given testosterone injections for 4 wk to produce serum concentrations equal to those of younger men. During testosterone administration muscle strength (isokinetic dynamometer) increased in both right and left hamstring and quadricep muscles as did the fractional synthetic rate of muscle protein (stable-isotope infusion). Ribonuclease protection assays done on total RNA from muscle showed that testosterone administration increased mRNA concentrations of IGF-I and decreased mRNA concentrations of insulin-like growth factor binding protein-4. We conclude that increasing testosterone concentrations in elderly men increases skeletal muscle protein synthesis and strength. This increase may be mediated by stimulation of the intramuscular IGF-I system. PMID- 7491932 TI - Route of nutrient delivery affects insulin sensitivity and liver glucose transporter expression in rat. AB - To optimize artificial nutrition (AN) techniques for patients suffering from malnutrition or reduced intestinal absorption, utilization of energy fuels, especially glucose, requires better understanding. Because the liver plays a key role in glucose homeostasis, the aim of this study was to assess the effects of continuous intragastric and intravenous nutrition on insulin secretion and several markers of liver glucose metabolism, especially glucose transporter GLUT 2. Wistar male rats underwent catheterization of either stomach (intragastric) or vena cava (intravenous) and received 24 h/day the same all-in-one formula over 7 to 14 days. The metabolic parameters from intragastrically fed rats did not differ significantly from those from orally fed control rats. Intravenous nutrition induced insulin resistance (marked hyperinsulinemia and/or mild hyperglycemia) and reduced liver GLUT-2 protein and mRNA levels. The decrease in liver GLUT-2 gene expression might be mediated either by an inhibitory role of hyperinsulinemia or by the decrease in gut or portal factors. These results suggest that the route of nutrient delivery influences their utilization by the liver. PMID- 7491933 TI - Contribution of fructose and lactate produced in placenta to calculation of fetal glucose oxidation rate. AB - We examined the rate of production of [14C]fructose and [14C]lactate from [U 14C]glucose by the placenta and the contribution of 14CO2 from fetal oxidation of these metabolic products to the calculation of glucose oxidation rate in fetal sheep. During fetal tracer infusions (n = 16), oxidation of fructose contributed 16 +/- 3% of total fetal CO2 production; oxidation of lactate accounted for 3.3 +/- 0.1%. Thus 80% of total fetal CO2 production resulted from direct oxidation of carbon atoms in glucose; the "direct" glucose oxidation fraction was 0.46 +/- 0.04. During maternal tracer infusion (n = 15), CO2 production from fructose was 21 +/- 3, 20 +/- 3, and 30 +/- 4% and from lactate was 16 +/- 3, 13 +/- 3, and 11 +/- 4% in hypo-, normo-, and hyperglycemic animals, respectively; the direct glucose oxidation fraction was 0.40 +/- 0.04, not different from the fraction obtained with the fetal tracer infusion. Fetal oxidation of substrates derived from glucose metabolism in the placenta contributes significantly to fetal CO2 production. Fetal oxidation of placental products of a metabolic substrate tracer should be considered in studies of fetal oxidative metabolism. PMID- 7491934 TI - Effect of hyperthyroidism on circulating levels of free and total IGF-I and IGFBPs in rats. AB - Thyroid hormones are suggested to have a permissive role in growth hormone (GH) and insulin-like growth factor I (IGF-I) action and a specific effect on plasma levels of some of the GH-independent IGF binding proteins (IGFBPs). We have investigated the effect of thyroxine (T4) administration on circulating levels of free and total (extractable) IGF-I and IGFBPs in 8-wk-old rats treated with 0, 200, 400, and 600 micrograms/kg T4, respectively. Serum free IGF-I was determined by an ultrafiltration method, serum total IGF-I after acid-ethanol extraction, and serum IGFBPs using Western ligand blotting, which yielded four distinct molecular bands: two single bands at 24 and 30 kDa and a double band at 38 and 42 kDa (38-42 kDa). After 13 days of hyperthyroidism, serum total IGF-I and the high molecular 38-42 kDa IGFBP were unchanged, whereas the 24-kDa IGFBP and 30-kDa IGFBP increased significantly (P < 0.05). Serum free IGF-I was significantly (P < 0.05) decreased in animals treated with 400 and 600 micrograms/kg T4. In addition, free IGF-I correlated inversely (P < 0.005) with the 24-kDa IGFBP, 30 kDa IGFBP, and serum total triiodothyronine. We conclude that hyperthyroidism in rats increases the circulating low-molecular IGFBPs and induces a reduction in free IGF-I. This may provide an important regulation of IGF bioactivity by thyroid hormones. PMID- 7491935 TI - Attenuation of glycogenolytic action of activin A in intact rat liver. AB - Activin A stimulates glucose production by causing glycogenolysis in isolated hepatocytes. To determine the physiological significance of this effect, we examined the effect of activin A on glucose production in the perfused liver. Unlike the effect in isolated cells, activin A did not enhance glucose production nor did it cause radiocalcium efflux in the perfused liver. There was no effect of activin A in the liver perfused in the opposite direction. Although activin A did not promote glucose production, it was recovered from the hepatic vein in a bioactive form. When liver perfusion was performed in partially hepatectomized rats, activin A increased radiocalcium efflux. In isolated hepatocytes, activin A increased inositol phosphates, and the effect of activin A was attenuated by the plasma membrane fraction of hepatocytes. The inhibitory effect of the plasma membrane was abolished by digestion of the membrane with trypsin. These results indicate that the effect of activin A on glucose production is attenuated in the intact liver and that a protein factor(s) in plasma membrane may be involved in the inhibition. PMID- 7491936 TI - Interleukin-1 beta-induced anorexia and pyrexia in rat: relationship to hypothalamic neuropeptide Y. AB - We investigated the effect of recombinant human interleukin-1 beta (rhIL-1 beta) induced anorexia and pyrexia on the hypothalamic neuropeptide Y (NPY)-ergic system, which stimulates feeding and reduces thermogenesis. In meal-fed rats, food intake decreased by 83%, 90 min after IL-1 beta treatment (1.3 micrograms/100 g ip; n - 8) vs. controls. NPY concentrations were significantly higher in the medial preoptic area (MPO), paraventricular (PVN), ventromedial (VMN), and dorsomedial (DMN) nuclei but unchanged in the arcuate nucleus (ARC) in both IL-1 beta-treated and pair-fed groups. Indomethacin (0.25 mg/100 g ip) reduced IL-1 beta-induced anorexia and tended to normalize NPY concentrations. In study 2, IL-1 beta increased core temperature by 1.1 degrees C above preinjection values (P < 0.001) and significantly raised NPY concentrations in the MPO, PVN, VMN, and DMN compared with controls, 60 min postinjection. Indomethacin prevented the pyrexia and normalized hypothalamic NPY levels. As NPY concentrations were not increased in the ARC (the hypothalamic site of synthesis), we suggest that the increased NPY levels may result from blocked release, which would be in accord with the known experimental effects of NPY. PMID- 7491937 TI - Reversion of insulin resistance in the rat during late pregnancy by 72-h glucose infusion. AB - To determine whether sustained exaggerated hyperinsulinemia in normoglycemic rats modifies insulin responsiveness during pregnancy, 17-day-pregnant and virgin rats were studied after receiving a continuous intravenous infusion (35 ml/day) of either 50% glucose or bidistilled water (controls) for 72 h. Plasma glucose was unchanged, whereas insulin was highly increased, and the effect was more marked in pregnant than in virgin rats. Insulin responsiveness, estimated under the hyperinsulinemic euglycemic clamp with 0.8 IU insulin.h-1.kg-1, was lower in control pregnant than in virgin rats but higher in pregnant than in virgin rats in those that had received the glucose infusion. The tissue glucose utilization metabolic index (GUI) was estimated with 2-deoxy-D-[1-3H]glucose in the clamped rats. The GUI was lower in heart, white- and red-fiber skeletal muscle, and adipose tissue in control pregnant rats than in control virgin rats, and, although the glucose infusion decreased that index in both red-fiber muscle and adipose tissue in virgin rats, glucose increased the index in red-fiber muscle in pregnant rats to the level found in virgin controls. Results therefore show that, when unaccompanied by hypoglycemia, sustained exaggerated hyperinsulinemia decreases insulin responsiveness in virgin rats but reverts insulin resistance in late-pregnant rats. PMID- 7491938 TI - Regulation of Ca(2+)-conducting currents in parathyroid cells by extracellular Ca(2+) and channel blockers. AB - High extracellular Ca2+ concentrations ([Ca2+]o) produce sustained intracellular Ca2+ responses in parathyroid cells that correlate with suppression of parathyroid hormone release. Using whole cell patch clamping, we identified two types of Ca(2+)-conducting currents in these cells. Type 1 currents were enhanced by raising [Ca2+]o and blocked by Cd2+ and nifedipine, whereas type 2 currents were resistant to blockade by these agents. Both types of membrane currents were cation nonselective, voltage independent over a broad range of membrane potentials, and blocked by the trivalent ions La3+ and Gd3+ (> 98%). Cd2+, La3+, and Gd3+ had biphasic effects on membrane conductance (Gm). At submicromolar concentrations, these ions increased Gm, whereas at higher concentrations they reduced Gm. In contrast to ionic channel blockers, nifedipine had only an inhibitory effect on the Ca(2+)-conducting currents that were sensitive to changes in [Ca2+]o (dose inhibiting 50% of maximal response = approximately 3-10 x 10(-8) M). Microfluorimetric ratio-imaging analysis of single parathyroid cells loaded with fura 2 showed that Gd3+ inhibited sustained intracellular Ca2+ responses to high [Ca2+]o. These findings suggest that the Ca(2+)-conducting currents identified in these studies may play a role in regulating intracellular Ca2+ responses in this system. PMID- 7491939 TI - Effect of meal timing on diurnal rhythm of human cholesterol synthesis. AB - To test whether the diurnal rhythm of cholesterol synthesis in humans is entrained to meal timing, the effect of a 6.5-h delay of mealtimes was investigated in four normal lipidemic male subjects. Cholesterol fractional synthetic rate was measured by deuterium incorporation from body water using blood sampling every 2 h. The baseline was a 24-h control period in which three Western-style meals were consumed at 0700, 1150, and 1640, followed by 3 days in which meals were delayed by 6.5 h, i.e., meals consumed at 1330, 1820, and 2310 without changing the sleep-wake and light-dark cycles. Cholesterol synthesis was maximal at 2200 +/- 0200 and minimal at 1130 +/- 0050 on the baseline day. On day 1 of the shifted meals, the maximum was delayed 6.0 +/- 0.5 h and the nadir was not changed. On day 3, the maximum was delayed 8.6 +/- 3.7 h and the minimum was delayed 6.5 +/- 2.4 h from baseline. The mean amplitude of the cholesterol rhythm was significantly greater on day 3,233 +/- 35%, compared with baseline which was 109 +/- 15%. A strong negative correlation (r = -0.66 +/- 0.10) was found between the rhythms of cholesterol synthesis and cortisol during the baseline day, but there was a phase delay in the rhythm of cholesterol synthesis relative to cortisol on day 1 and day 3. Findings indicate that the 24-h variation in cholesterol synthesis is strongly dependent on meal timing. PMID- 7491940 TI - Urea cycle intermediate kinetics and nitrate excretion at normal and "therapeutic" intakes of arginine in humans. AB - We investigated the effects of a high dietary supplement of arginine on plasma arginine, ornithine, and leucine kinetics and on urea production and excretion in five healthy young adult men. Subjects received either 56 or 561 mg arginine.kg 1.day-1 for 6 days via a complete L-amino acid diet, and on day 7 a tracer protocol (first 3 h fasted; next 5 h fed) was conducted, involving primed constant intragastric infusions of L-[15N2-guanidino,5,5-2H2]arginine, L-[5 13C]ornithine, L-[5,5,5-2H3]leucine, and [15N2]urea, with a prime of H13CO3. Plasma arginine and ornithine fluxes increased significantly (P < 0.05) with arginine supplementation, as did the rate of conversion of plasma labeled arginine to ornithine (P < 0.05) and rate of ornithine oxidation (P < 0.001). However, absolute changes in ornithine kinetics were less than those for arginine or those based on changes expected from the change in arginine intake, implying a complex compartmentation in both whole body arginine and ornithine metabolism. The plasma NO3 concentration, daily output of total NO3, and conversion of [15N]arginine to NO3 did not differ between the diets. Urea production and excretion were reduced significantly with arginine supplementation, suggesting an anabolic effect on the whole body nitrogen economy, possibly via the raised plasma insulin levels (P = 0.013) during the prandial phase. PMID- 7491941 TI - Human GLUT-2 overexpression does not affect glucose-stimulated insulin secretion in MIN6 cells. AB - Accumulated evidence suggests that GLUT-2, in addition to its role in glucose transport, may also have other functions in glucose-stimulated insulin secretion. As a first step in addressing this possibility, we have engineered MIN6 cells overexpressing human GLUT-2 by transfection with human GLUT-2 cDNA. Stable transformants harboring human GLUT-2 cDNA exhibited an approximately twofold increase in 3-O-methyl-D-glucose uptake at 0.5 and 15 mM. Glucokinase activity or glucose utilization measured by conversion of [5-3H]glucose to [3H]H2O was not, however, altered in the MIN6 cells overexpressing human GLUT-2. Furthermore, glucose-stimulated insulin secretion was not affected by over-expression of human GLUT-2. An abundance of GLUT-2, therefore, does not correlate with the glucose responsiveness of cells in which glycolysis is regulated at the glucose phosphorylating step. These data suggest that GLUT-2 by itself does not have significant functions other than its role in glucose transport in glucose sensing by MIN6 cells. PMID- 7491942 TI - PACAP stimulates catecholamine release from adrenal medulla: a novel noncholinergic secretagogue. AB - The secretory response of the adrenal medulla to pituitary adenylate cyclase activating polypeptide (PACAP), a novel polypeptide with 68% structural homology to vasoactive intestinal polypeptide (VIP), was investigated in anesthetized rats by in vivo microdialysis. The injection of PACAP (1.5 nmol) caused a greater amount of increase in catecholamine concentration than carbachol (30 nmol) or VIP (30 nmol) in the dialysate for a period of 60 min. The ratios of norepinephrine to epinephrine in the dialysate after stimulation with these compounds showed no change from resting conditions. The simultaneous application of both nicotinic and muscarinic antagonists (10 mM mecamylamine and 1 mM atropine) eliminated the increase in catecholamine secretion induced by carbachol. In contrast, PACAP induced catecholamine secretion was not inhibited by these cholinergic antagonists. These data led to the following conclusions: 1) PACAP was more potent than either carbachol or VIP in enhancing the secretion of catecholamine from the adrenal medulla, and 2) PACAP-induced catecholamine secretion was due to the direct action of PACAP on the adrenal medulla rather than an indirect action mediated by acetylcholine release. These results strongly suggest that PACAP is a noncholinergic secretagogue of the adrenal medulla in rats. PMID- 7491944 TI - Osteogenic protein-1 enhances phenotypic expression in ROS 17/2.8 cells. AB - Osteogenic protein-1 (OP-1) stimulates bone morphogenesis in vivo and modulates osteoblast growth and differentiation in vitro. Treatment of ROS 17/2.8 cells with OP-1 resulted in a time- and concentration-dependent inhibition of [3H]thymidine incorporation. In contrast, OP-1 treatment stimulated phenotypic differentiation in ROS 17/2.8 cells, as indicated by enhanced 1) alkaline phosphatase activity (4-fold); 2) alkaline phosphatase mRNA (5-fold); 3) parathyroid hormone receptor mRNA (2-fold), and 4) parathyroid hormone-stimulated adenosine 3',5'-cyclic monophosphate accumulation (2-fold). OP-1-induced changes in cell growth and gene expression were sensitive to cycloheximide and actinomycin D. Measurement of [3H]thymidine incorporation and alkaline phosphatase activity in situ revealed heterogeneity in the cellular responses to OP-1. Proliferating cells exhibited less alkaline phosphatase activity than nonproliferating cells, whereas cells expressing high levels of alkaline phosphatase incorporated little [3H]thymidine. Our data delineating the responses of mature differentiated osteoblasts to OP-1 suggest that potentiation of osteoblast differentiated function is an important component of bone morphogenesis in vivo. PMID- 7491943 TI - Effects of protein, methionine, or chloride on acid-base balance and on cysteine catabolism. AB - Metabolism of cysteine to sulfate results in production of H+, whereas metabolism of cysteine to taurine does not. Rats were fed a basal low-protein diet or a diet with excess protein, methionine, or chloride for 2-3 wk, and effects of these treatments on acid-base homeostasis and on cysteine metabolism were determined. Hepatocytes from rats fed diets with excess methionine, but not from rats fed diets with excess protein or chloride, catabolized a high proportion of cysteine to taurine (32% vs. 4-7% for other groups), and intact rats fed excess methionine excreted more sulfur as taurine (51% of total sulfur vs. 1-6% for other groups). The formation of taurine vs. sulfate as the end product of cysteine catabolism provides a metabolic compensation that minimizes the acid load in rats fed excess sulfur amino acids. However, increased production of taurine vs. sulfate is not a general adaptive response to acidogenic diets. PMID- 7491945 TI - Glucocorticoid deprivation alters in vivo glucose uptake by muscle and adipose tissues of GTG-obese mice. AB - The effect of 1 wk of glucocorticoid deprivation by surgical adrenalectomy (ADX) on tissue 2-deoxy(-)[U-14C]glucose (2-DG) uptake and hepatic glucose production (HGP) was assessed in conscious, catheterized mice 5 wk after the induction of obesity with gold thioglucose (GTG). Despite the prevailing hyperglycemia and hyperinsulinemia, glucose uptake by heart, quadriceps muscle, and interscapular brown adipose tissue (BAT) of GTG-obese mice was unchanged compared with controls, suggesting that the hyperglycemia of GTG-obese mice is able to compensate for the insulin resistance of these tissues. In contrast, epididymal white adipose tissue (WAT) of GTG-obese mice showed increased glucose uptake with hyperglycemia and hyperinsulinemia. ADX decreased the hyperglycemia and lowered the elevated glycogen content of the liver of GTG-obese mice. ADX reduced glucose uptake by heart and WAT of control and GTG-obese mice, consistent with the concomitant decrease in insulinemia. Glucose uptake by muscle of control and GTG obese mice was not significantly decreased after ADX despite the decrease in insulin, and ADX increased glucose uptake by BAT of GTG-obese mice, suggesting increased sympathetically mediated thermogenesis in this tissue. HGP was increased in GTG-obese mice compared with controls, and ADX significantly reduced HGP in both GTG-obese and control mice. These results suggest that the improved glucose tolerance of ADX GTG-obese mice and ADX control mice is due to a decrease in HGP rather than an increase in peripheral glucose uptake. PMID- 7491946 TI - Suppression of circulating calcitriol and duodenal active Ca transport by ketoconazole in pregnant rats. AB - Active Ca transport in the duodenum and the circulating level of calcitriol are elevated during pregnancy and lactation in the rat. Because calcitriol stimulates Ca transport in nonmated rats, we investigated its contribution to the increased transport during pregnancy and lactation. Rapid suppression of calcitriol from 28 +/- 3 to 8 +/- 0.4 pg/ml with the steroid hydroxylase inhibitor ketoconazole resulted in a 34% suppression of Ca transport in nonmated rats. At the end of pregnancy, when calcitriol concentration was suppressed from 64 +/- 7 to 12 +/- 2 pg/ml, the transport ratio decreased by 44%. Ca transport did not correlate with calcitriol levels between 40 and 80 pg/ml, suggesting a threshold level for maximal Ca transport stimulation. During lactation at even higher calcitriol levels, ketoconazole treatment again resulted in marked reduction in calcitriol from 124 +/- 1 to 71 +/- 12 pg/ml, but without any concurrent reduction in Ca transport in the duodenum. We conclude that in the vitamin D-replete rat the pregnancy-mediated, and probably also the lactation-mediated, increase in active Ca transport capacity is dependent on an increase in circulating calcitriol up to a certain threshold level. PMID- 7491948 TI - Use of R-beta-[1-11C]hydroxybutyrate in PET studies of regional cerebral uptake of ketone bodies in humans. AB - A method for determining regional cerebral utilization of ketone bodies in humans is described. After a bolus injection of R-beta-[1-11C]hydroxybutyrate, the time course of the tracer in the brain was measured with positron emission tomography in five healthy volunteers. The regional cerebral blood flow was measured separately. The tracer uptake in the brain could be well described by a single rate constant, indicating that the concentration of unmetabolized ketone bodies in the brain is very low and that transport across the blood-brain barrier is the rate-limiting step. At an average plasma concentration of beta-hydroxybutyrate of 0.043 mumol/ml, the utilization rate was estimated to be 0.48 nmol.ml-1.min-1. In accordance with previous animal studies, the utilization rate was found to increase almost linearly with increasing plasma concentration of beta hydroxybutyrate. Furthermore, the utilization was higher in gray than in white matter. Finally, the ratio between the utilization in the basal ganglia and the brain as a whole was lower for ketone bodies than for glucose. PMID- 7491947 TI - Phospholipase A2 activity is increased in guinea pig uterine cervix in late pregnancy and at parturition. AB - Release of arachidonic acid from phospholipids by phospholipases A2 (PLA2) is the rate-limiting step in prostaglandin (PG) synthesis. PGE2 and PGF2 alpha are essential intermediates in interstitial collagenase-mediated degradation of type I collagen, a key step in cervical dilatation at parturition. We demonstrate that PLA2 is present in cytosolic fractions of guinea pig cervix and PLA2 activity is increased during cervical dilatation at parturition. In the cervix of nonpregnant animals, PLA2 activity against phosphatidylcholine (PC) and phosphatidylethanolamine (PE) was 13 +/- 6 and 49 +/- 27 pmol.min-1.mg protein-1, respectively. Levels were similar at day 25 of pregnancy. At day 50, PLA2-PC increased to 190 +/- 54 pmol.min-1.mg-1, and PLA2-PE rose to 592 +/- 127 pmol.min 1.mg-1. At parturition (68 +/- 2 days) there were further increases of 49-67%. PLA2 activity declined toward basal levels 2 days postpartum. Almost 50% of the enhanced cytosolic PLA2 (cPLA2) activity at day 50 or at parturition was of high molecular mass and was identified as the "85-kDa cPLA2." Increases in cPLA2 activity at these time points were not, however, associated with increases in cPLA2 protein or phosphorylation of cPLA2, compared with nonpregnant animals. This study suggests that multiple PLA2 enzymes, including cPLA2, are involved in cervical dilatation at parturition. PMID- 7491949 TI - Constant-pressure perfusion of rat hindlimb shows alpha- and beta-adrenergic stimulation of oxygen consumption. AB - Isolated rat hindlimbs were perfused at 37 degrees C and constant physiological pressure (80 +/- 0.5 mmHg) while the flow rate that was allowed to freely self adjust was monitored. Under these conditions, evidence was obtained for both alpha- and beta-adrenergic stimulation of oxygen consumption (VO2) in contrast to constant-flow perfusion, which has only convincingly shown alpha-adrenergic stimulation of VO2 in response to adrenergic agents. Addition of norepinephrine (NE; 1-33 nM) led to an increase in VO2 with a maximum of 29% above the basal value at 3.3 nM, even though the flow rate decreased. Phenylephrine (3.3-33 nM) and vasopressin (10-100 pM) also showed similar, but lesser in magnitude, vasoconstriction-associated stimulatory effects on VO2. Prazosin (an alpha 1 antagonist) completely reversed the NE-mediated decrease in flow rate and significantly blocked the increased VO2. In contrast, isoproterenol (10-1,000 nM) increased both flow rate (30%) and VO2 (32%). The isoproterenol-stimulated VO2 was not blocked by the beta 1-, beta 2-antagonist propranolol (10 microM), although the increased flow was reversed. In the presence of propranolol (1 or 10 microM), BRL-35135A (a beta 3-agonist) also stimulated VO2 (18%) without significant change in flow rate. These results lend further support to the role of the alpha 1-adrenoceptor in muscle VO2. In addition there is evidence for the presence of a functional beta 3-adrenoceptor as an additional subtype responsible for NE-mediated thermogenesis in the rat hindlimb. PMID- 7491950 TI - Recording of a large-conductance chloride channel in normal rat lactotrophs. AB - Membrane current fluctuations resembling channel openings and closings were observed in the whole cell configuration of the patch-clamp technique in normal rat lactotrophs in primary culture. Using high-gain head stage in whole cell configuration, we characterized the nature and pharmacological properties of the ionic channel underlying these fluctuations. This channel, found in small numbers (< 10 per cell), was specific for Cl- because its reversal potential varied with Cl- gradients, according to the Nernst equation, and its unitary amplitude was linearly related to membrane potential from -100 to 0 mV. Slope conductance was close to 100 pS. Analyzing open times, we demonstrated its Ca2+ and potential dependence. Four sublevels were observed. We suggest that this channel, belonging to the background Cl- channel group, takes part in the regulation of intracellular Cl- concentration of normal rat lactotrophs. PMID- 7491951 TI - Insulin-like growth factor I accelerates protein synthesis in skeletal muscle during sepsis. AB - Sepsis causes an inhibition of protein synthesis in gastrocnemius that is resistant to the anabolic effects of insulin. The purpose of the present studies was to investigate the effect of recombinant human insulin-like growth factor I (IGF-I) on protein synthesis during a 30-min perfusion of the isolated rat hindlimb from septic rats. Inclusion of IGF-I (1 or 10 nM) in the perfusate stimulated protein synthesis in gastrocnemius of septic rats 2.5-fold and restored rates of protein synthesis to those observed in control rats. The stimulation of protein synthesis did not result from an increase in the RNA content but was correlated with a 2.5-fold increase in the translational efficiency. The enhanced translational efficiency was accompanied by a 33 and 55% decrease in the abundance of free 40S and 60S ribosomal subunits, respectively, indicating that IGF-I accelerated peptide-chain initiation relative to elongation/termination. These studies provide evidence that IGF-I can accelerate protein synthesis in gastrocnemius during chronic sepsis by reversing the sepsis induced inhibition of peptide-chain initiation. PMID- 7491952 TI - Mucin gene structure and expression: protection vs. adhesion. AB - The cloning of mucin cDNAs brought about by the application of molecular biology and molecular analyses constitutes a major step in understanding mucin structure and function. Here two classes of mucins are described: epithelium-associated and endothelium/leukocyte-associated mucins, which have thus far been described separately in the literature. The epithelial mucins are generally believed to play a role in cytoprotection. The endothelial and leukocyte class of mucins are adhesion molecules involved in lymphocyte homing and lymphocyte activation or are part of the adhesion cascade that plays a role in the initiation of inflammation. Mucins in general contain many threonine and serine residues, which are extensively O-glycosylated. Due to this profound glycosylation, mucins have a filamentous conformation. By virtue of their extended filamentous, and often negatively charged, structure, mucins can act as a barrier protecting the cell. However, when an opposing cell has specific receptors for mucins, adhesion can override the barrier function. Therefore, mucins may be powerful two-edged swords: they are both protective and adhesive. PMID- 7491953 TI - Cholecystokinin receptors. AB - The cholecystokinin (CCK) and gastrin families of peptides act as hormones and neuropeptides on central and peripheral CCK receptors to mediate secretion and motility in the gastrointestinal (GI) tract in the physiological response to a normal meal. CCK and its receptors are also widely distributed in the central nervous system (CNS) and contribute to the regulation of satiety, anxiety, analgesia, and dopamine-mediated behavior. Although the wide distribution, myriad number of functions, and reported pharmacological heterogeneity of CCK receptors would suggest a large number of receptor subtypes, the application of modern molecular biological techniques has identified two CCK receptors, CCK-A receptor (CCK-AR) and CCK-B receptor (CCK-BR), that mediate the actions of CCK and gastrin; gastrin receptors have been found to be identical to CCK-BR. CCK-AR, found predominantly in the GI system and select areas of the CNS, have high affinity for CCK and the nonpeptide antagonist L-364,718, whereas CCK-BR, found predominantly in the CNS and select areas of the GI system, have high affinity for CCK and gastrin and the nonpeptide antagonist L-365,260. Both CCK-AR and CCK BR are highly conserved between species, although there is some tissue-specific variation in expression. Recombinant receptor expression faithfully reproduces the native receptor pharmacology and signal transduction pathways, allowing direct comparisons of receptor function between species as well as serving as a convenient source of receptor. Our present knowledge of the chromosomal localization, receptor gene structure, and primary sequence will allow further studies in disease association, receptor regulation, and structure-function analysis. PMID- 7491954 TI - Ca(2+)-dependent amylase secretion from SLO-permeabilized rat pancreatic acini requires diffusible cytosolic proteins. AB - Streptolysin O (SLO)-permeabilized pancreatic acini are now frequently used to study regulated exocytosis in the exocrine pancreas. In this paper we introduce alpha-toxin as a possible alternative permeabilization agent to SLO. Both alpha toxin and SLO are bacterial cytolysins, but the membrane pores generated by SLO are approximately 5-10 times larger than those formed by alpha-toxin. The Ca2+ requirements for amylase secretion from both types of permeabilized acini were identical, maximal amylase secretion being obtained at 30 microM Ca2+ with an effective concentration of approximately 3-4 microM Ca2+ producing 50% of the maximal response. However, Ca(2+)-stimulated amylase secretion from the SLO permeabilized acini stopped after 10-15 min, unlike secretion from the alpha toxin-permeabilized cells, which continued for at least 50 min. The rapid cessation of secretion from the SLO-treated acini reflects the rapid decline in the responsiveness of the cells observed after permeabilization. This decline in Ca(2+)-dependent secretion appears to be due to the loss of cytosol, since addition of purified rat brain cytosol to nonresponsive SLO-permeabilized acini reconstituted regulated secretion. Because alpha-toxin-permeabilized acini maintained their responsiveness, the cytosolic factors lost from the SLO permeabilized cells must be retained within the toxin-treated cells. The reconstitutive activity of the brain cytosol was nondialyzable but heat and trypsin sensitive, suggesting that the factors responsible are proteins. Of the cytosols screened (brain, liver, spleen, muscle, and lacrimal) only those prepared from brain or lacrimal gland reconstituted Ca(2+)-dependent amylase secretion.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7491955 TI - Influence of gravity and body position on normal oropharyngeal swallowing. AB - The influence of gravity, if any, on pharyngeal bolus transport is unknown. The aim of the present study was to evaluate the effect of gravity and body position on the radiological and manometric events during the normal oropharyngeal swallow. In eight healthy male volunteers, we performed simultaneous videoradiography and manometry, using a manometry catheter incorporating a perfused sleeve measuring upper esophageal sphincter pressure and three solid state transducers recording pharyngeal pressures. Swallows of 5 and 10 ml of barium were performed in three different body positions: upright, horizontal, and 30 degrees head down (inverted). Hypopharyngeal intrabolus pressure increased significantly in the horizontal and inverted positions compared with upright, P = 0.0001. This resulted in increased maximal sphincter diameters during bolus flow (P = 0.0001) and shorter duration of sphincter opening (P = 0.03). As a result, transsphincteric flow increased in the horizontal and inverted positions, P = 0.04. Total swallowing duration, oral and pharyngeal transit time, pharyngeal peristaltic amplitude and duration, the length of the bolus in the pharynx, and excursions of the hyoid and larynx were unaffected by body position. We conclude that intrabolus pressure is an important determinant of upper esophageal sphincter opening in the healthy oropharynx and that gravity does not influence pharyngeal bolus transport. The healthy upper esophageal sphincter has residual opening capacity that can be demonstrated by altering body posture. PMID- 7491956 TI - Direct G protein activation reverses impaired CCK signaling in human gallbladders with cholesterol stones. AB - Human gallbladders were used to investigate the mechanisms of the impaired contraction induced by cholecystokinin (CCK) associated with cholesterol stones. Single muscle cells were isolated enzymatically with collagenase. Inositol 1,4,5 trisphosphate was measured by high-performance liquid chromatography. Diacylglycerol was assayed by thin-layer chromatography. CCK stimulation showed decreased muscle contraction and production of inositol 1,4,5-trisphosphate and diacylglycerol in gallbladders with cholesterol stones compared with those with pigment stones. Exogenous calmodulin induced maximal contraction of 22.4 +/- 0.5 and 21.0 +/- 0.6% in gallbladders with cholesterol and pigment stones, respectively. Similar findings were observed with a synthetic diacylglycerol analogue. Two G protein activators, aluminum fluoride and guanosine 5'-O-(3 thiotriphosphate), evoked similar responses in these two types of gallbladders, with maximal contractions of 21.3 +/- 0.4 and 23.3 +/- 0.5%, respectively, in those with cholesterol stones and 20.9 +/- 0.8 and 22.6 +/- 0.4%, respectively, in those with pigment stones. These results suggest that receptor-dependent ligands like CCK cannot fully activate the intracellular pathways, which, however, can be fully stimulated by circumventing receptors with G protein activators or second messengers. After G protein activation, the pathways appear to be functionally intact. The defect might then reside in the receptor or in the interaction between receptors and G proteins. PMID- 7491957 TI - Na(+)-K(+)-ATPase alpha 1- and beta 1-mRNA and protein levels in rat small intestine in experimental ileitis. AB - Na(+)-K(+)-adenosinetriphosphatase (ATPase) plays a key role in the absorption of electrolytes, water, and nutrients from the small intestine. The expression of Na(+)-K(+)-ATPase was examined in isolated enterocytes during the course of the ileal inflammatory response elicited by intraluminal administration of 2,4,6 trinitrobenzenesulfonic acid. The ileal inflammatory response was characterized by a marked cellular infiltrate, villous atrophy, and crypt hyperplasia along with fibrosis and smooth muscle hypertrophy. Peak levels of myeloperoxidase were observed at day 7, and ileal mucosal injury was paralleled by increases in ileal mucosal permeability. Ileal enterocytes were harvested from days 3 to 30 after the induction of ileitis. Decreases in Na(+)-K(+)-ATPase functional activity were observed from days 3 to 21 and were accompanied by corresponding decreases in Na(+)-K(+)-ATPase pump abundance, alpha 1- and beta 1-protein expression, and mRNA abundance, whereas Na(+)-K(+)-ATPase turnover, Michaelis-Menten constant values, and inhibition constant values for Na+ and ouabain, respectively, were unaltered. Alterations in transcriptional and posttranscriptional events may determine the changes in Na(+)-K(+)-ATPase activity in this particular model. Additionally observed increases in thymidine kinase and ornithine decarboxylase activities appear to signify alterations in the state of differentiation of the ileal epithelium and may determine the phenotypic expression of enterocyte transporters and permeability in the setting of inflammation. PMID- 7491958 TI - EGF inhibits secretagogue-induced cAMP production and amylase secretion by Gi proteins in pancreatic acini. AB - In pancreatic acinar cells, the epidermal growth factor (EGF) receptor interacts with both cholera toxin- and pertussis toxin (PTX)-sensitive G proteins. In the present study, isolated rat pancreatic acini were used to investigate the effect of EGF on basal and secretagogue-induced adenosine 3',5'-cyclic monophosphate (cAMP) production and amylase release. EGF increased cAMP production and amylase release in pancreatic acini. However, cAMP accumulation and amylase release elicited by either vasoactive intestinal peptide (VIP) or forskolin were inhibited by EGF (17 nM). EGF inhibited the VIP-induced cAMP production and amylase release with a half-maximal effective concentration of 3 and 2 nM, respectively. EGF had no effect on the N6,2'-O-dibutyryladenosine-3',5' monophosphate-stimulated amylase release, suggesting that the inhibitory effect of EGF on the VIP- and forskolin-induced cAMP production is due to inhibition of adenylyl cyclase. PTX pretreatment of the acini led to an increase of the basal, EGF-, and VIP-stimulated cAMP accumulation and amylase release, indicating that PTX-sensitive G proteins exert tonic inhibition of adenylyl cyclase even in the absence of agonist. In PTX-pretreated acini, the inhibitory effect of EGF on the VIP-induced cAMP production and amylase release was abolished. In conclusion, these results suggest that EGF inhibits secretagogue-induced cAMP production via activation of PTX-sensitive G proteins in rat pancreatic acini, whereas EGF induced cAMP production and amylase release occurs via a PTX-insensitive pathway. PMID- 7491959 TI - Activation of MAP kinase and translocation with HSP27 in bombesin-induced contraction of rectosigmoid smooth muscle. AB - We have investigated whether mitogen-activated protein (MAP) kinase cascade is essential for sustained contraction of smooth muscle cells of the rabbit rectosigmoid. We have identified MAP kinase as one of the enzymes activated by bombesin, performed immunologic studies blocking the activation of MAP kinase, and conducted confocal localization of MAP kinase in relation to heat-shock protein (HSP27), postulated to be involved in the sustained contraction of smooth muscle. Immunoblotting revealed two forms of MAP kinase (42 and 44 kDa). Activation of MAP kinase by bombesin was rapid, reaching a maximum in 30 s and subsequently declining. [D-Phe6,Leu13,psi(CH2NH),Phe14]BN-(6-14), a potent bombesin antagonist, and protein kinase C (PKC) inhibitors 1-(5 isoquinolinylsulfonyl)-2-methylpiperazine, calphostin C, and chelerythrine inhibited the increase in MAP kinase induced by bombesin. Immunofluorescent dual labeling and confocal microscopy indicate that these two proteins are closely distributed in resting cells and that during bombesin-induced contraction MAP kinase translocates accompanied by HSP27. In conclusion, a series of events involving PKC activation, MAP kinase activation, and MAP kinase-HSP27 translocation could be the signaling pathway involved in bombesin-induced sustained contraction. PMID- 7491960 TI - Intermittence of blood flow in liver sinusoids, studied by high-resolution in vivo microscopy. AB - Kupffer cell migration and leukocyte-vessel wall interactions cause temporary slowing and/or stoppage of blood flow through individual liver sinusoids. Such temporal heterogeneity of flow was quantified in anesthetized mice and rats. Video recordings of red blood cell flow in 44 networks containing 8-16 sinusoids each were analyzed for 5- to 10-min periods. Flow was graded "fast," "slow," "stopped," or "reversed" based on red blood cell velocity. The mean numbers of flow changes (between grades) per minute in zone 1 vs. zone 3 were 1.39 vs. 0.78 (mouse) and 1.25 vs. 0.09 (rat). The mean percentage of time for each flow grade differed significantly between zones 1 and 3 and between species. For example, fast flow was present in zone 1 sinusoids for 51% of the time in mice and for 74% in rats; in zone 3 the corresponding numbers were 76 and 95%. Flow stasis was present in zone 1 sinusoids for 19% of the time in mice and for 7% in rats; in zone 3 the corresponding numbers were 2 and 0%. Thus considerable intermittence of perfusion exists, and the flow conditions create very different microenvironments for hepatocytes in zone 1 vs. zone 3. PMID- 7491961 TI - Gastrin receptor antagonist YM022 prevents hypersecretion after long-term acid suppression. AB - Female rats were treated orally for 13 wk with YM022 (300 mumol.kg-1.day-1) and with omeprazole (400 mumol.kg-1.day-1) or famotidine (900 mumol.kg-1.day-1) with or without YM022. At 2 h after the last dose, YM022 and omeprazole markedly inhibited basal and pentagastrin-induced acid secretion. Famotidine was less potent than YM022 and omeprazole against both secretions. The degree of increase in plasma gastrin level in the three groups was parallel to the antisecretory potencies of the drugs. At 14 days after the cessation of omeprazole treatment, the secretory response to pentagastrin increased above that of the control. This hyperresponse lasted for > or = 56 days. In the famotidine-treated group, a small increase in secretory response to pentagastrin was observed but was not statistically significant. The increase in secretory response to pentagastrin was paralleled by an increase in mucosal cell mass. In contrast, YM022 not only exhibited a long-lasting inhibition of pentagastrin-induced acid secretion but also prevented the hyperresponse to pentagastrin caused by omeprazole. These results indicate that the hypergastrinemia caused by long-term administration of antisecretory drugs increases mucosal secretory response to pentagastrin through a gastrin/cholecystokinin B receptor-mediated pathway in rats. PMID- 7491962 TI - Haustral septations increase axial and radial distribution of luminal contents in glass models of the colon. AB - We used two glass models of the colon to test the hypothesis that luminal septations increase efficiency of flow. Each was a straight glass tube 4 cm in diameter and filled with water. One had four septations narrowing the lumen to 2 cm; the other had no septations. For each run, liquid dye and one solid test particle were placed at one end of the model. A single pressure pulse was applied to the dye and particle. The distance each traveled was compared using analysis of variance. The nonseptated model had poor mixing of dye, and the particle moved only short distances. In the septated model, there was thorough mixing of dye and the particle moved rapidly along what appeared as a central core of high-velocity liquid flow from one septal opening to another. Compared with the nonseptated model, this produced statistically significant increases in the distances traveled by means of 32 (dye) and 90% (particle). Our studies suggest that septations in a tubular organ facilitate intraluminal mixing and flow of liquids and solids. PMID- 7491963 TI - Signal-transduction pathways causing slow synaptic excitation in guinea pig myenteric AH neurons. AB - Intracellular recordings were obtained from myenteric AH neurons of guinea pig ileum in vitro. Slow excitatory synaptic responses associated with decreased potassium conductance (gK), inhibition of the spike afterhyperpolarization current (AHC), and increased chloride conductance (gCl) were mimicked by senktide, a neurokinin3 receptor agonist. Intracellular guanosine 5'-O-(3 thiotriphosphate) (GTP gamma S) decreased gK and increased gCl irreversibly after nerve stimulation or senktide application. Myenteric neurons in pertussis toxin (PTX)-treated tissues responded normally to senktide and nerve stimulation. Forskolin and phorbol 12,13-dibutyrate (PDBu) inhibited gK and the AHC but did not activate gCl. The AHC was not reduced by subthreshold concentrations of forskolin (10 nM) or PDBu (3 nM) alone but was inhibited by forskolin and PDBu applied together. Inhibitors of phospholipase C (D-609) or protein kinases (staurosporine) reduced slow synaptic and senktide responses. The protein phosphatase inhibitor, calyculin A, caused an inward current, a decrease in gK, and AHC inhibition but did not activate gCl. We conclude that slow excitatory synaptic responses are mediated by PTX-insensitive G proteins and activation of phospholipase C and protein kinases. Forskolin and PDBu activate pathways that inhibit gK. The mechanisms for activation of gCl are unknown. PMID- 7491964 TI - Tonic suppression of gastric acid secretion by endogenous peptides in neonatal rats. AB - Stimulation of gastric acid secretion by secretagogues was measured in developing rats by in vivo and in vitro techniques. Basal acid outputs in vivo were very low in 8- and 14-day-old rats compared with those in 20- and 30-day-old rats. In 20 day-old rats, all secretagogues increased acid output in vivo, whereas only carbachol, pentagastrin, and sulfated cholecystokinin octapeptide (CCK-8S) were active in 14-day-old rats. In contrast, basal acid output in vitro and stimulation by secretagogues did not differ significantly with age. CCK-8S stimulated acid output in vitro in 14-day-old rats was blocked by L-365,260, L 364,718, tetrodotoxin, and atropine, but not by hexamethonium, whereas gastrin stimulated acid output was blocked only by L-365,260. Furthermore, acid output in vivo was elevated three- to fourfold by subcutaneous naloxone-methiodide or L 364,718, but not by L-365,260, in 14-day-old rats; none of these antagonists produced an effect in 20-day-old rats. These studies show that low basal gastric acid output in neonatal rats is caused by tonic inhibitory regulation by endogenous regulatory peptides. PMID- 7491965 TI - Expression of somatostatin receptor genes and their role in inhibiting Cl- secretion in HT-29cl.19A colonocytes. AB - Recent studies in a cultured model of the intestinal epithelium (HT-29cl.19A) have shown that somatostatin-14 (SS-14) inhibits the Cl- secretory process by acting at multiple G protein-dependent sites. These actions may underlie the antidiarrheal properties of SS peptides. This study has investigated the expression of specific SS receptor subtypes (SSTR) in HT-29cl.19A and examined their role in mediating SS antisecretory actions. Two predominant SSTR, SSTR1 and SSTR2, were identified by reverse transcriptase-polymerase chain reaction (RT PCR) of mRNA from polarized HT-29cl.19A monolayers. Receptor binding studies showed evidence of two distinct populations of binding sites consistent with the known properties of SSTR1 and SSTR2. The role of SSTR in inhibition of secretion was investigated by comparing the effectiveness of native and synthetic SS peptides on adenosine 3',5'-cyclic monophosphate (cAMP)-dependent Cl- secretion. Secretion stimulated by the receptor-mediated agonist prostaglandin E2 (PGE2) was inhibited > 70% by SS-14 with a 50% effective concentration (EC50) of 32 nM. In contrast, SMS-201-995 (SMS) and RC-160 exhibited little or no antisecretory activity (maximum inhibition of 15 +/- 1.9 and 2.8 +/- 1.9%, respectively, at 100 microM; EC50 > 1.5 microM). Similar effects on PGE2-stimulated cAMP accumulation were also observed. SS-14, but not SMS, also inhibited secretion stimulated by dibutyryl cAMP, which acts independently of changes in cellular cAMP. Pretreatment with pertussis toxin reversed the antisecretory effects of SS peptides.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7491966 TI - Role of gastric blood flow in impaired defense and repair of aged rat stomachs. AB - To study impaired gastric mucosal tolerance against noxious agents in aged rats, possible factors underlying this observation were compared in anesthetized Fisher 344 young and aged rats. The gastric mucosa was damaged by in situ exposure to 80% ethanol for 30-45 s and by 1 M NaCl for 10 min followed by saline (pH = 1.0) for 60 min in chambered stomachs. The lesion area was significantly larger and epithelial restitution was significantly slower in aged than in young rats after both types of injury. Changes in gastric blood flow were monitored by laser Doppler velocimetry. Young, but not aged, rats showed a marked increase in gastric blood flow in response to 1 M NaCl, acid challenge, and 640 microM capsaicin for 60 min. Young rats showed a higher density of calcitonin gene related peptide (CGRP)-staining nerve fibers around submucosal blood vessels and higher mucosal release of prostaglandin E2 and leukotriene C4 than did aged rats. These data suggest that impaired mucosal defense and reduced restitution in aged rats is related to lack of hyperemic response caused by mucosal injury and H+ back-diffusion, which is probably due to decreased density of CGRP-staining nerve fibers and prostaglandin biosynthetic capacity in the mucosa. PMID- 7491967 TI - Change in hepatic handling of hepatocyte growth factor during liver regeneration in rats. AB - The change in the hepatic uptake clearance of hepatocyte growth factor (HGF) during liver regeneration in rats was examined. The hepatic uptake clearance per gram liver of 125I-labeled HGF in vivo decreased to a minimum level only 15 min after partial hepatectomy (PH). In the perfused liver, in which the effect of endogenous substances can be ignored, the hepatic uptake clearance of 125I-HGF did not change until at least 3 h after PH, suggesting that the decrease in the clearance in vivo is caused by some endogenous inhibitors. Such a decrease in the clearance cannot be explained only by the elevated endogenous HGF concentration in plasma. In addition, at 48 h after PH the decrease in hepatic uptake clearance was comparable for in vivo and perfused liver (approximately 50 and 60% reduction, respectively), suggesting that the decreased hepatic clearance in vivo at that time was caused mainly by reduction in the binding and/or uptake by the liver itself. In addition, the hepatic uptake clearance of 125I-HGF at that time in the presence of an excess (135 pM) of unlabeled HGF was comparable with that of the control, indicating that only the saturable portion of hepatic uptake clearance is reduced. The internalization rate constant representing the probability of cell surface-bound 125I-HGF being internalized per unit time increased to three times that of the control at 15 min after PH, demonstrating that the internalization of HGF is enhanced immediately after the start of liver regeneration. PMID- 7491968 TI - Transport and steady-state accumulation of putrescine in brush-border membrane vesicles of rabbit small intestine. AB - Absorption of polyamines from the lumen is essential for cell proliferation in small intestine but also in other rapidly growing body tissues and tumors. Intestinal uptake of polyamines is thought to involve one or more transport systems, but the characteristics of these systems have not yet been clearly elucidated. Because high levels of putrescine have been identified in intestinal lumen, we explored kinetic, physiochemical, and structural features of uptake of this diamine across rabbit intestinal brush-border membrane vesicles (IBBMV) prepared by CaCl2 or MgCl2 precipitation procedure. Initial rates of putrescine influx were measured during 5-min incubations at 25 or 37 degrees C (optimal temperature) for concentrations of 0.45-145 microM. At both temperatures, kinetics of putrescine transport fitted a model with a single Michaelis-Menten uptake component plus a nonsaturable uptake component. At 37 degrees C, the kinetic parameters for the saturable component of putrescine uptake, Km,app and Vmax,app, were 16.8 +/- 4.7 microM and 19.9 +/- 2.8 pmol.mg protein-1.min-1, respectively. The value of the constant for the nonsaturable component of putrescine uptake (P = 0.45 +/- 0.06 x 10(-8) l.mg protein-1.s-1) suggested this component represented essentially nonspecific binding of putrescine to IBBMV. Cadaverine, spermidine, and spermine were competitive inhibitors of putrescine transport, with inhibition constants equal to 47, 117, and 219 microM, respectively. When effects of a variety of alkyldiamines and structural analogues of polyamines (1 mM) on influx of 5.6 microM putrescine were compared, cadaverine, methylglyoxal bis(guanylhydrazone) (MGBG), and cyclic derivatives of MGBG were found to exhibit the highest inhibitory potencies.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7491970 TI - La3+ and pH sensitivity of Ca2+ entry and intracellular store filling in gastric parietal cells. AB - The fluorescent Ca2+ indicator fura 2 was used to measure cytosolic free [Ca2+] ([Ca2+]i) in order to obtain information about relative rates of Ca2+ influx into parietal cells during treatment with carbachol (a cholinergic agonist) or thapsigargin (TG, a Ca(2+)-mobilizing agent) or during reloading of the internal Ca2+ stores. In Ca(2+)-containing solutions, carbachol-, TG-, and reloading stimulated Ca2+ entry exhibited nearly identical sensitivity to La3+ [inhibition constant (Ki) approximately 10 microM] or low pH (pKi approximately 7.0). In experiments in which carbachol and TG were used, there was no additional increase in [Ca2+]i when TG was added to carbachol-treated cells or when carbachol was added to cells previously treated with TG. Thus it is likely that a single Ca2+ entry pathway serves a signaling function as well as a role in refilling the Ca2+ store during reloading. Because the Ca2+ pathway is exquisitely sensitive to pH and serosal pH increases during stimulant-induced H+ secretion (which is activated by increases in [Ca2+]i), this mechanism will exert positive feedback on parietal cells in the intact stomach. When parietal cells were pretreated with carbachol in Ca(2+)-free solutions, reloading was independent of pH and La3+, suggesting that Ca(2+)-containing solutions should be used to determine the properties of the influx pathway. PMID- 7491969 TI - Gastric antisecretory and anti-ulcer actions of IL-1 in rat involve different IL 1 receptor types. AB - Limited knowledge exists concerning the interleukin-1 (IL-1) receptor type (IL 1RT) mediating the potent antisecretory and gastro-protective actions of IL-1. In the present study, the gastric actions of IL-1 beta and two related mutant proteins, yIL-1 beta delta 4, an analogue that preferentially binds to IL-1-RTII, and mutant yIL-1 beta N7/Q, an analogue that has equal affinity as IL-1 beta for IL-1RTI and IL-1RTII, have been compared. Modulation of IL-1 gastric actions were also investigated using monoclonal antibody (MAb) preparations raised against IL 1RTI or IL-1RTII. In the pylorus-ligated rat, yIL-1 beta delta 4, yIL-1 beta N7/Q, and IL-1 beta (all at 1 microgram/kg ip) reduced gastric acid secretion (50, 79, and 78%, respectively), indicating the importance of IL-1RTII binding for antisecretory activity. This was further substantiated in experiments using the MAb preparations, which showed that IL-1 beta (1 microgram/kg ip) antisecretory activity was reversed by MAb IL-1RTII (10-50 micrograms/kg sc) but not by MAb IL-1RTI (50 micrograms/kg sc). In contrast, at dosages 10-fold higher (10 micrograms/kg ip) than that used in the study to inhibit acid secretion, IL-1 beta and yIL-1N7/Q equally reduced (approximately 80%) indomethacin-induced gastric damage, but yIL-1 beta delta 4 was ineffective. The results using yIL-1 beta delta 4 indicated that impairment of IL-1RTI binding capacity appeared to be paralleled by a decreased gastroprotective effect.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7491971 TI - Halothane and octanol block Ca2+ oscillations in pancreatic acini by multiple mechanisms. AB - This study has investigated halothane and octanol, reported inhibitors of gap junction permeability, for their effects on acinar cell intracellular Ca2+ concentration ([Ca2+]i) signaling. Halothane and octanol alone at maximal concentrations induced a sustained rise in [Ca2+]i of 23 +/- 4 and 29 +/- 5 nM, respectively. Cholecystokinin (CCK, 20 pM) induced [Ca2+]i oscillations in single acinar cells within the acinus to a peak of 275 +/- 17 nM, rising from a basal level of 55 +/- 3 nM. These oscillations were completely abolished by superfusion with both halothane (4 mM) and octanol (1 mM), concentrations that blocked the spread of Lucifer yellow from cell to cell within an acinus. Lower concentrations of octanol markedly reduced the oscillation frequency (0.2 and 0.5 mM octanol: reduction in oscillation frequency of 69 +/- 6 and 43 +/- 6%, respectively). These agents however, over the same concentration range, also exhibited similar inhibitory effects on [Ca2+]i oscillations in single cells dispersed from the acinus (reduction in oscillation frequency of 75 +/- 10 and 32 +/- 12% for 0.2 and 0.5 mM octanol, respectively), suggesting additional effects other than on gap junctions. Halothane inhibited inositol 1,4,5-trisphosphate [Ins(1,4,5)P3] production in response to both 1 and 10 nM CCK (31 and 40% inhibition, respectively), possibly explaining its effects on [Ca2+]i oscillations, whereas octanol showed no significant inhibition. Octanol, unlike halothane, blocked Ins(1,4,5)P3-induced Ca2+ release from permeabilized acini, an effect that was most pronounced at a more physiological Ins(1,4,5)P3 concentration. Octanol did not affect Ins(1,4,5)P3 binding to Ins(1,4,5)P3 receptor preparation. In conclusion, although halothane and octanol block gap junction permeability in pancreatic acinar cells, these agents also affect Ins(1,4,5)P3 production and Ca2+ mobilization in response to agonist stimulation. PMID- 7491972 TI - NO modulates the apicolateral cytoskeleton of isolated hepatocytes by a PKC dependent, cGMP-independent mechanism. AB - Nitric oxide (NO) induces smooth muscle relaxation. We examined whether NO or its mediator of this action, guanosine 3',5'-cyclic monophosphate (cGMP), similarly induces relaxation of the apicolateral cytoskeleton in hepatocytes. Apical (canalicular) contractions were observed in isolated rat hepatocyte couplets by videomicroscopy, tight junction permeability was determined in the couplets by paracellular penetration of Texas red-dextran, and cytosolic Ca2+ (Ca2+i) was measured in isolated hepatocytes by fluorescence imaging. Unexpectedly, the NO donor sodium nitroprusside potentiated rather than inhibited apical contraction, in a cGMP-independent manner. This action of nitroprusside was blocked by hemoglobin or by inhibition of protein kinase C (PKC). Nitroprusside and 3 morpholinosydnonimine, another NO donor, each increased the permeability of hepatocyte tight junctions, a known effect of PKC in this cell type, and induced translocation of that kinase to the plasma membrane, as determined by immunocytochemistry. Neither nitroprusside nor dibutyryl cGMP changed the amplitude or frequency of Ca2+i signals in hepatocytes. Exogenous NO thus modulates the apicolateral cytoskeleton of hepatocytes via PKC activation rather than via cGMP or Ca2+i. These observations suggest a new role for NO: to activate PKC. PMID- 7491973 TI - Role of CFTR in chloride secretion across human tracheal epithelium. AB - We have tested two hypotheses: 1) the cystic fibrosis transmembrane conductance regulator (CFTR) represents the predominant Cl conductance in the apical membrane of human tracheal epithelium, and 2) CFTR in this tissue is close to maximally activated under baseline conditions. In support of the first hypothesis, we found 1) when the level of differentiation of cultures was varied by varying the culture conditions, there was a significant positive correlation between the levels of CFTR and the magnitude of mediator-induced Cl secretion. 2) Amiloride insensitive baseline short-circuit current (Isc) and mediator-induced increases in Isc were inhibited by diphenylamine-2-carboxylic acid (DPAC) but not by 4,4' diisothiocyanostilbene-2,2'-disulfonic acid (DIDS), a pharmacology consistent with passage of apical membrane Cl current through CFTR; Ca-activated Cl channels are inhibited by DIDS but not by DPAC. 3) Raising temperature from 22 degrees to 37 degrees C increased 125I efflux, and this increase was inhibited by DPAC and blockers of protein kinase A, but not by DIDS or 1,2-bis(2-aminophenoxy)ethane N,N,N',N'-tetraacetic acid-acetoxymethyl ester. In support of the second hypothesis, we have earlier shown [M. Yamaya, W.E. Finkbeiner, S.Y. Chun, and J.H. Widdicombe. Am. J. Physiol. 262 (Lung Cell. Mol. Physiol. 6): L713-L724, 1992] that adenosine 3',5'-cyclic monophosphate (cAMP)-elevating agents are essentially without effect on Isc across primary cultures of human tracheal epithelium. Here, we further show that these agents are also usually without effect on 125I efflux; the mean increase in efflux in response to elevating cAMP was approximately 20% that of raising temperature from 22 degrees to 37 degrees C. PMID- 7491974 TI - Phosphatidylcholine molecular species of calf lung surfactant. AB - This paper reports the detailed composition of molecular species of the phosphatidylcholines (PCs) in pulmonary surfactant from calves. PC isolated by thin-layer chromatography (TLC) was converted to benzoylated diradyl glyceride derivatives, which were separated by TLC according to linkage group. Quantification of linkage groups by analysis of total fatty acid content demonstrated that surfactant PC contained 97.2% diacyl, 2.4% alkyl-acyl, and 0.4% alkenyl-acyl compounds. The diacyl and alkyl-acyl diglyceride derivatives were separated into individual molecular species by high-performance liquid chromatography. Four major species constituted 87% of the diacyl compounds. Dipalmitoyl phosphatidylcholine (DPPC) was the most abundant constituent, contributing 41% of the total PC. A second disaturated species, palmitoyl myristoyl phosphatidylcholine (PMPC), also contributed an additional 12% of total PC. At least 65% of PMPC occurred as the 1-palmitoyl-2-myristoyl/isomer, which has a lower melting point than the 1-myristoyl-2-palmitoyl compound. These results show that most of pulmonary surfactant PC is a relatively simple mixture, that numerous minor compounds are present in small but possibly important amounts, and that in surfactant from calves, the widely reported estimate that DPPC constitutes 60% of surfactant PC is too large by 50%. PMID- 7491975 TI - Hypoxia inhibits L-arginine uptake by pulmonary artery endothelial cells. AB - Under physiological conditions, L-arginine transport by porcine pulmonary artery endothelial cells (PAEC) is mediated by system y+, a sodium-independent transport system that accounts for 60 +/- 5% of L-arginine transport, and system Bo,+, a sodium-dependent system that accounts for 40 +/- 5% of transport. Because NO production is dependent on intracellular L-arginine content and intracellular L arginine content depends on transport of extracellular L-arginine, we examined the effect of hypoxia on L-arginine transport and intracellular L-arginine content in PAEC. Exposure of passage 3-7 PAEC in monolayer culture to 0% O2 for 4 h decreased L-arginine transport via system y+ from 120 +/- 10 to 81 +/- 23 (in pmol.mg protein-1.30 s-1) (P < 0.001), whereas 20-h exposures decreased transport from 122 +/- 17 to 84 +/- 18 (P < 0.001) in system y+ and from 104 +/- 19 to 90 +/- 26 (P < 0.05) in system Bo,+. Exposure to 5% O2 for 3-5 wk decreased L arginine transport via system y+ from 128 +/- 15 to 73 +/- 13 (P < 0.001) and via system Bo,+, from 105 +/- 25 to 65 +/- 13 (P < 0.001). Kinetic studies revealed that hypoxia decreased the maximal transport velocity but not the apparent Michaelis constant for both system y+ and system Bo,+, and the decreases in transport were not reversible after return to normoxia for up to 24 h. Long-term exposure, i.e., 3-5 wk, to 5% O2 also resulted in decreases in intracellular L arginine content (0.75 +/- 0.10 vs. 0.49 +/- 0.09 nmol/10(6) cells, P < 0.05) which did not reverse after return to normoxia for 24 h.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7491976 TI - Hypoxia inhibits L-arginine synthesis from L-citrulline in porcine pulmonary artery endothelial cells. AB - Both non-arginine-depleted and arginine-depleted pulmonary artery endothelial cells (PAEC) actively convert citrulline into arginine. Exposure to hypoxia for 4 24 h inhibited arginine synthesis from citrulline in intact cells and in cell homogenates. The conversion of L-citrulline to L-argininosuccinate by argininosuccinate synthetase (AS) was inhibited by exposure to hypoxia for 4, 12, or 24 h. The conversion of argininosuccinate to arginine by argininosuccinate lyase was inhibited by exposure to hypoxia for 24 h but not for 4-12 h. The decrease of L-arginine biosynthesis during hypoxia coincided with the increase of intracellular glutamine content and was abrogated by preventing an increase in intracellular glutamine. In addition, AS activity was inversely related to glutamine content in the medium. These results indicate that hypoxia inhibited the L-arginine biosynthetic pathway via decreased activity of AS. The latter is related to increased glutamine content. Hypoxic inhibition of arginine synthesis from citrulline did not result in a decrease of arginine content, suggesting that PAEC are able to maintain intracellular arginine for up to 24 h despite reduction in the L-arginine biosynthetic pathway. PMID- 7491977 TI - Activation of NF-kappa B and elevation of MnSOD gene expression by thiol reducing agents in lung adenocarcinoma (A549) cells. AB - The effect of reducing agents, including N-acetylcysteine (NAC), dithiothreitol (DTT), and 2-mercaptoethanol (2-ME) on nuclear transcription factor-kappa B (NF kappa B) activation and manganese superoxide dismutase (MnSOD) expression was investigated in a pulmonary adenocarcinoma (A549) cell line. NAC, DTT, and 2-ME each activated the transcription factor NF-kappa B and increased steady-state levels of MnSOD mRNA and enzyme activity in these cells. In addition, NAC, DTT, and 2-ME increased chloramphenicol acetyltransferase (CAT) activity in cells transfected with a construct containing the CAT gene under the control of the rat MnSOD promoter. SOD and catalase (500 U/ml) plus ethanol (1 mM) did not inhibit activation of NF-kappa B or elevation of steady-state MnSOD mRNA levels by NAC, DTT, or 2-ME. Controls in which comparable amounts of O2-. to those produced by thiols were generated by hypoxanthine and xanthine oxidase, or in which H2O2 was added directly, had neither activated NF-kappa B nor elevated MnSOD mRNA. This shows that reactive oxygen intermediates, which may be formed during autooxidation, may not contribute to activation of NF-kappa B. Because the MnSOD promoter also contains potential binding sites for other transcription factors, such as promoter-selective transcription factor-1 (SP-1), activator protein-1 (AP 1), AP-2, adenosine 3',5'-cyclic monophosphate-regulator element binding factor (CREB), and transcription factor IID complex (TFIID), the effect of thiols on their activation also were evaluated. In contrast to findings with NF-kappa B, there was only minor activation of AP-1 by thiols, and none of the other transcription factors were activated by thiols. AP-1 activation was inhibited by catalase (500 U/ml) plus SOD plus ethanol (1 mM). Addition of 700 microM H2O2 also activated AP-1, and catalase at 500 U/ml prevented this activation. This indicates that H2O2 produced as a result of autooxidation of thiols can activate AP-1 but not NF-kappa B. Thus a close association between exposure to reducing agents, activation of NF-kappa B, and elevation of MnSOD gene expression is demonstrated. PMID- 7491978 TI - Sequence of rat surfactant protein A gene and functional mapping of its upstream region. AB - Surfactant protein A (SP-A) is the major pulmonary surfactant protein. We have isolated a rat SP-A genomic clone and determined the sequence from 2.9 kilobases upstream of the transcriptional start through the termination of translation. The exon-intron structure of the rat gene has been determined and compared with the mouse, rabbit, and human genes. We have localized the major transcriptional start site in adult rat lung to nucleotide 30 downstream from the start of the TATA box. Functional mapping indicates that a DNA fragment containing 163 nucleotides upstream of the transcriptional start (-163) can function as a promoter of transcription of a reporter gene in both lung and nonlung derived cell lines. However, the function of this element is weaker in cells of nonlung origin. DNA elements located between -2902 and -163 silence the promoter activity in both lung and nonlung cells. Because the SP-A gene promoter region exhibits limited tissue specificity, the results suggest the existence of other DNA elements which overcome the silencer and confer further lung specificity. PMID- 7491979 TI - Antioxidant-surfactant liposomes mitigate hyperoxic lung injury in premature rabbits. AB - Surfactant liposomes, encapsulating CuZn-superoxide dismutase (SOD) and catalase, increase alveolar type II cell antioxidant activity and protect cells against oxidant stress. We examined whether intratracheal instillation of antioxidant surfactant liposomes increases lung antioxidant activity in premature rabbits. Pregnant New Zealand White rabbits were delivered by cesarean section on day 28 or 29 of gestation or allowed to deliver spontaneously. After premature birth or at 2 days of age in the term rabbits, the pups from each litter were divided into four groups. One group received 0.1 ml/15 g birth wt of antioxidant-surfactant liposomes by intratracheal injection and was then exposed to hyperoxia (> 95% oxygen) for 24 h and killed. The second group received an equal amount of surfactant liposomes without antioxidant enzymes and was exposed to hyperoxia for 24 h. The third group received air placebo and was exposed to hyperoxia for 24 h, and the fourth group was killed after birth if premature or at 2 days of age if term. After the pups were killed, lung homogenates were investigated for total SOD and catalase activity and DNA content. Each treatment group consisted of 12 15 rabbit pups. Lung antioxidant enzyme activity increased with advancing maturity. Among the premature rabbits, total lung SOD and catalase activity were lowest in the pups killed before hyperoxia and the air placebo controls exposed to hyperoxia, intermediate in the pups treated with liposomes without antioxidant enzymes and hyperoxia, and highest in the pups that received antioxidant surfactant liposomes and hyperoxia.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7491980 TI - In vivo lipopolysaccharide pretreatment inhibits cGMP release from the isolated perfused rat lung. AB - Administration of bacterial lipopolysaccharide (LPS) to rats stimulates synthesis of nitric oxide (NO), a free radical molecule that activates soluble guanylate cyclase, thereby increasing intracellular guanosine 3',5'-cyclic monophosphate (cGMP) concentration and inducing systemic vasodilatation. To investigate the effect of endotoxemia on the pulmonary NO/cGMP signal transduction system, we measured the release of cGMP by isolated-perfused lungs of rats that received an intraperitoneal injection of LPS (1 mg/kg) or saline 2 days earlier. Over 90 min, 1.4 +/- 0.78 and 0.079 +/- 0.016 nmol cGMP accumulated in pulmonary perfusates of saline- and LPS-treated rats, respectively (P < 0.05). Despite addition to the perfusate of Zaprinast, superoxide dismutase, or A23187, markedly less cGMP was released from the lungs of rats exposed to LPS than from the lungs of control rats. In contrast, after ventilation with 100 parts per million NO gas, cGMP accumulating in the perfusate of the lungs of both groups of rats was markedly increased, and the quantity of cGMP released from the lungs of LPS-treated rats was similar to that released by control rat lungs (2.8 +/- 0.57 vs. 3.3 +/- 0.88 nmol, P = NS). With the use of immunoblot techniques, equal concentrations of constitutive endothelial NO synthase were detected in the lungs of rats treated with saline or LPS. These results demonstrate that the NO/cGMP signal transduction system is abnormal in the lungs of rats exposed to LPS, at least in part, at the level of endothelial NO synthase activation. PMID- 7491981 TI - Dietary changes improve survival of CFTR S489X homozygous mutant mouse. AB - Over 90% of untreated CFTR S489X homozygous (CF) mutant mice reportedly die of intestinal obstruction by 40 days of age, significantly limiting their usefulness as a model for the human disease. Because the period of highest mortality is during the week after weaning, we hypothesized that providing a low-residue liquid diet would improve survival and growth. When 99 CF mice that survived to 10 days of age were fed Peptamen (Clintec Nutrition), an elemental liquid diet, and housed on corn-cob bedding, 88% of them survived to maturity (50 days). The diet causes only minor histologic and ion transport changes in the intestines of normal mice and does not reduce growth rate or size. CF mice raised on Peptamen continue to display severe pathological changes in the intestine and completely lack a adenosine 3',5'-cyclic monophosphate-inducible chloride current in the cecum. This combination of dietary and bedding changes provides a reliable method for keeping CF mice alive well into adulthood and will be useful for the evaluation of the effect and duration of potential therapies for CF. PMID- 7491982 TI - Tachykinins induce gelatinase production by guinea pig alveolar macrophages: involvement of NK2 receptors. AB - To determine whether tachykinins induce gelatinase production by guinea pig alveolar macrophages (AM), and to characterize the mechanism involved, we incubated AM with substance P (SP), neurokinin A (NKA), or the NH2-terminal fragment of SP, SP(1-7). The effects of increasing concentrations of selective NK1 and NK2 agonists on tachykinin-induced gelatinase production were also evaluated, as were the effects of a selective NK2 antagonist. Gelatinase activity in conditioned culture media (CCM) was assessed by zymography and quantified by image analysis. SP increased 92-kDa gelatinase activity in CCM of AM in a concentration-dependent manner, with a maximum increase at 10(-4) M. NKA, the NH2 terminal fragment of SP, and an NK1-selective agonist had no effect. In contrast, a selective NK2 agonist induced a concentration-dependent increase in gelatinase activity. The increase in this activity induced by SP and the selective NK2 agonist was inhibited by a selective NK2 antagonist. We conclude that SP induces gelatinase production by AM through NK2 receptor activation. The release of gelatinase may constitute one mechanism through which SP contributes to the epithelial lesions observed in bronchial hyperreactivity and asthma. PMID- 7491983 TI - Potential role of NADH oxidoreductase-derived reactive O2 species in calf pulmonary arterial PO2-elicited responses. AB - Our laboratory has previously reported evidence that tone responses of isolated endothelium-removed calf pulmonary arteries elicited by changes in PO2 appear to be mediated via changes in H2O2 and guanosine 3',5'-cyclic monophosphate, and that the PO2 sensor mechanism is hypothesized to involve changes in superoxide anion (O2-.) production by a microsomal NADH-oxidoreductase, which is the major source of O2-. detected by lucigenin-elicited chemiluminescence (CL) in this tissue. In this study we examined if the flavoprotein-directed inhibitor of O2-. producing NAD(P)H oxidoreductases, diphenyliodonium (DPI), could be employed as an inhibitor of O2-. production by NADH oxidoreductase, which functions as a selective probe for PO2-elicited tone responses in calf pulmonary arterial smooth muscle. It was found that 1 microM DPI inhibited NADH-dependent production of CL in the arterial smooth muscle homogenate by 49% (n = 10). DPI reduced basal CL from endothelium-removed pulmonary arteries by 41% (n = 15). In endothelium removed pulmonary arteries precontracted with U-46619, the hypoxic contraction of 2.3 +/- 0.5 g was reduced to 0.1 +/- 0.4 g (n = 7) by DPI, and the reoxygenation relaxation of 32.7 +/- 7.5% was decreased to 4.4 +/- 1.4% (n = 7). DPI did not have any significant effect on U-46619- or K(+)-elicited tone generation. DPI also did not alter the relaxation to H2O2 (1 microM-0.1 mM, n = 6), nitric oxide (0.42 nM-420 nM, n = 12), or isoproterenol (1 nM-1 microM, n = 6).(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7491984 TI - Mechanism of impaired beta-adrenoceptor responsiveness in atopic sensitized airway smooth muscle. AB - Decreased airway relaxation to beta-adrenoceptor stimulation has been hypothesized as a potential mechanism leading to enhanced bronchoconstrictor responsiveness in asthma. In addressing potential mechanisms underlying this phenomenon, the relative contributions of beta-adrenoceptor-coupled transmembrane signaling mechanisms were examined in isolated rabbit tracheal smooth muscle (TSM) passively sensitized with serum from atopic asthmatic patients and in TSM comparably exposed to non-atopic (control) human serum. During half-maximal isometric contraction of the tissues with acetylcholine, relative to control TSM, the sensitized tissues exhibited significant attenuation of both their maximal relaxation (Rmax) and sensitivity (i.e., -log 50% Rmax) to cumulative administration of isoproterenol (P < 0.001) or prostaglandin (PG)E2 (P < 0.001). In contrast, the relaxation responses to forskolin, a diterpene that directly activates adenylate cyclase, were similar in both tissue groups. Extended studies demonstrated that the attenuated relaxation to isoproterenol and PGE2 in sensitized TSM was 1) ablated by pretreatment with the muscarinic M2-receptor antagonists methoctramine (10(-6) M) or gallamine (10(-4) M); 2) also inhibited by pretreatment with pertussis toxin (100 ng/ml), which ADP ribosylates the inhibitory G protein (G(i)) negatively coupled to adenylate cyclase activation; and 3) associated with diminished adenosine 3',5'-cyclic monophosphate accumulation in response to isoproterenol administration. Moreover, based on Western immunoblot analysis, we found that G(i) protein expression was increased in membrane fractions from sensitized TSM, related to enhanced expression of the G(i) alpha 3 subunit. Collectively, these observations provide new evidence that the impaired beta-adrenoceptor-mediated relaxation in atopic sensitized airways is associated with increased muscarinic M2 receptor/G(i) protein-coupled expression and function. PMID- 7491985 TI - Intratracheal injection of endotoxin and cytokines. IX. Contribution of CD11a/ICAM-1 to neutrophil emigration. AB - Intratracheal injection of endotoxin [lipopolysaccharide (LPS)] in rats causes acute inflammation characterized by the emigration of neutrophils (PMNs) into the bronchoalveolar airspace. Antibody to PMN adhesion molecule CD11a inhibited LPS initiated PMN accumulation in bronchoalveolar lavage (BAL) fluid by 32% (P < 0.001). Antibody to the endothelial CD11a counterreceptor intercellular adhesion molecule-1 (ICAM-1) inhibited LPS-initiated PMN accumulation in BAL fluid by 66% (P < 0.0001). Combined antibody blockade of ICAM-1 and the C-X-C chemokine cytokine-induced neutrophil chemoattractant (CINC) inhibited LPS-initiated PMN emigration by 80%, significantly more than antibody against either ICAM-1 or CINC alone. To study the relative contribution of alveolar macrophages and PMNs to intra-alveolar tumor necrosis factor (TNF), the LPS-induced TNF in BAL fluid was measured after depletion of circulating PMNs with a cytolytic antibody to CD18. Although the anti-CD18 antibody completely abrogated LPS-initiated PMN emigration into BAL fluid, TNF levels in BAL fluid were unaffected, suggesting that alveolar macrophages are the predominant cellular source of LPS-induced TNF production. In conclusion, 1) CD11a, ICAM-1, and CINC play major roles in the LPS-initiated emigration of PMNs into the bronchoalveolar space, and 2) the TNF that drives ICAM-1 and CINC expression is derived largely from alveolar macrophages rather than PMNs. PMID- 7491986 TI - Enhanced growth of fetal and neonatal pulmonary artery adventitial fibroblasts is dependent on protein kinase C. AB - The earliest and most striking proliferative changes in the neonatal pulmonary arterial wall occur in the adventitia where the fibroblast resides. The protein kinase C (PKC) pathway is developmentally regulated and important in vascular cell growth. We tested the hypothesis that developmental differences in growth of pulmonary artery adventitial fibroblasts would be detectable in vitro and dependent on PKC. Fibroblasts were isolated from bovine fetal, neonatal, and adult pulmonary arteries. Growth was measured by [3H]thymidine incorporation and cell counts. Under serum-stimulated conditions, fetal and neonatal pulmonary artery fibroblasts grew faster and reached higher plateau densities than adult cells. Increased growth of fetal cells in vitro was dependent on time of harvest during fetal life (early > late). Under quiescent conditions, fetal and neonatal fibroblasts had increased DNA synthesis compared with adult cells in response to the PKC agonist phorbol 12-myristate 13-acetate. To test whether the developmental differences in fibroblast growth were dependent on PKC, three different inhibitor strategies were used (dihydrosphingosine, phorbol-ester induced downregulation, and heparin). Fetal and neonatal fibroblasts were more susceptible than adult cells to each antagonist strategy. Finally, we measured whole cellular PKC catalytic activity and found it correlated with growth and susceptibility to PKC inhibition (i.e., fetal PKC activity > neonatal > adult). We conclude that PKC-dependent developmental differences in growth of pulmonary artery fibroblasts are detectable in vitro and that the enhanced growth capacity of fetal and neonatal cells may contribute to the dramatic adventitial thickening seen in vivo after hypoxic exposure in the neonatal calf. PMID- 7491988 TI - Na+/H+ exchange in pulmonary artery smooth muscle from spontaneously hypertensive and Wistar-Kyoto rats. AB - Na+/H+ exchanger regulation of intracellular pH (pHi) may play a key permissive role in the mitogen-induced vascular smooth muscle cell growth that occurs in systemic and pulmonary vascular remodeling. Spontaneously hypertensive rats (SHR) have increased Na+/H+ exchange in systemic vessels as well as greater systemic vascular remodeling compared with normotensive Wistar-Kyoto rats (WKY). In contrast to WKY, SHR demonstrate only mild pulmonary hypertension and no increased remodeling to hypoxia compared with WKY. We therefore wondered whether Na+/H+ exchange in pulmonary artery smooth muscle (PASM) of SHR might not be elevated compared with WKY. Baseline pHi, Vmax, pK0.5, and Hill coefficient were compared in 12- to 14-wk-old SHR and WKY PASM and aortic smooth muscle (AoSM) segments by ratio fluorescence spectroscopy. The Vmax, pK0.5, and Hill coefficient were significantly increased in SHR AoSM segments compared with WKY AoSM segments (53, 0.55, and 53%, respectively; P < 0.05). There were no differences in these values between SHR and WKY PASM segments, unlike the AoSM segments. We conclude that the Na+/H+ exchanger activity in PASM in the SHR is the same as in the WKY, which is in contrast to systemic arteries where Na+/H+ exchange is greater in the SHR. PMID- 7491987 TI - Effects of separate and combined ETA and ETB blockade on ET-1-induced constriction in perfused rat lungs. AB - To evaluate the role of endothelin (ET) receptors in ET-1-induced pulmonary vasoreactivity, we studied the effects of ET-receptor agonists and antagonists in isolated perfused rat lungs. ET-1 (1-10 nM) caused concentration-dependent pulmonary vasoconstriction and gross pulmonary edema at a concentration of 10 nM. The combination of the selective ETA antagonist BQ-123 and the selective ETB antagonist BQ-788 inhibited ET-1-induced pulmonary vasoconstriction more effectively than BQ-123 alone, whereas BQ-788 alone enhanced the constriction. ET 1-induced hydrostatic pulmonary edema was prevented by the combination of BQ-123 and BQ-788 but not by either BQ-123 or BQ-788 alone. After the addition of 125 ng of exogenous ET-1, the perfusate levels of ET-1 were significantly higher in BQ 788-treated lungs than in either the vehicle control or BQ-123-treated lungs. The selective ETB agonist IRL-1620 also caused pulmonary vasoconstriction and edema, both of which were completely inhibited by BQ-788. ET-1-induced transient vasodilation was abolished by BQ-788 but was unaffected by BQ-123. These results suggest that in the isolated perfused rat lung, ET-1-induced vasoconstriction is mediated by both ETA and ETB receptors, whereas ET-1-induced transient vasodilation is mediated exclusively by the ETB receptor. Blockade of ETB receptors may result in enhanced ET-1 activity (via the ETA receptor) through inhibition of the ETB-mediated clearance of ET-1. Thus combined ETA and ETB blockade is required for the complete inhibition of ET-1-induced vasoconstriction in the rat pulmonary circulation. PMID- 7491989 TI - Putrescine uptake in hamster lung slices and primary cultures of type II pneumocytes. AB - Putrescine is accumulated in the lungs of various species by an active uptake system that also mediates the uptake of cystamine and paraquat. We have characterized this uptake in both lung slices and type II pneumocytes isolated from hamsters by trypsin digestion, differential adherence on plastic, and centrifugation on a discontinuous Percoll gradient. The accumulation of [14C]putrescine in lung slices was shown to be temperature and energy dependent, and to obey saturation kinetics, with mean calculated values of apparent Michaelis constant (Km) 29.4 microM and maximum rate of uptake (Vmax) 637 nmol.g 1.h-1. In the presence of cystamine or paraquat, the putrescine uptake was reduced in a manner compatible with competitive inhibition. The calculated inhibitor constants (Ki) were 16 and 1,017-1,328 microM for the inhibition by cystamine and paraquat, respectively. The cellular localization of [3H]putrescine in lung slices after incubation with 2.5 microM putrescine was determined by light-microscopic autoradiography. Labeling was present in type II and possibly also in type I pneumocytes of the alveolar epithelium but not in macrophages, endothelium, or cells of the interstitium. Two days after their isolation, cultured type II pneumocytes exhibited an uptake of putrescine that had kinetic characteristics similar to those of slices (Km of 23 microM and Vmax of 3.06 mumol.g protein-1.h-1) and was also competitively inhibited by paraquat (Ki of 222-350 microM paraquat). Our data demonstrate the presence of an active uptake system for putrescine in both lung slices and cultured type II pneumocytes.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7491991 TI - Mechanisms for the generation of gas-exchange surface area in rat lung. AB - During development, rat lung seems to make use of the mechanisms of 1) expansion, 2) replication, and 3) subdivision to increase its gas-exchange surface area. To quantitate the contribution of each one of these mechanisms from birth to adulthood, four models of lung development have been mathematically evaluated: 1) existing saccules at birth septate (subdivide) during the first 2 postnatal weeks, creating smaller alveoli that undergo a continuous expansion up to adulthood; 2) same as the previous model except that new alveoli are also formed by means other than septation after the second postnatal week; 3) same as the previous model except that the alveoli formed by means other than septation start forming immediately after birth; and 4) saccules continue to be formed up to adulthood and subsequently septate, forming alveoli. In an evaluation of these models mathematically, it was found that expansion followed by replication is the most important mechanism for the generation of gas-exchange surface area in all four models in the maturing lung. Although the contribution of septation is important during early postnatal life, its importance decreases with age. PMID- 7491990 TI - ETA-receptor antagonist prevents and reverses chronic hypoxia-induced pulmonary hypertension in rat. AB - The selective endothelin-A (ETA)-receptor antagonist BQ-123 has been shown to prevent chronic hypoxia-induced pulmonary hypertension in the rat. Therefore in the current study we utilized BQ-123 to test the hypothesis that blockade of the ETA receptor can reverse as well as prevent the increase in mean pulmonary artery pressure, right ventricle-to-left ventricle plus septum ratio, and percent wall thickness in small (50-100 microns) pulmonary arteries observed in male Sprague Dawley rats exposed to normobaric hypoxia (10% O2, 2 wk). Infusion of BQ-123 (0.4 mg.0.5 microliter-1.h-1 for 2 wk in 10% O2) begun after 2 wk of hypoxia significantly reversed the established pulmonary hypertension and prevented further progression of right ventricular hypertrophy during the third and fourth week of hypoxia. BQ-123 infusion instituted before exposure to hypoxia completely prevented the hypoxia-induced pulmonary hypertension, right ventricular hypertrophy, and pulmonary vascular remodeling. These findings suggest that, in the lung, hypoxia induced an increase synthesis of endothelin-1, which acts locally on ETA receptors to cause pulmonary hypertension, right heart hypertrophy, and pulmonary vascular remodeling, while ETA-receptor blockade can both prevent and reverse these processes. PMID- 7491992 TI - Human lung cell beta 2-adrenergic receptors desensitize in response to in vivo administered beta-agonist. AB - Few studies have addressed whether target tissue adrenergic receptors in humans undergo desensitization in response to agonist administration. To determine whether lung cell beta 2-adrenergic receptors (beta 2-AR) undergo such desensitization, we harvested bronchial epithelial cells and alveolar macrophages via bronchoscopy from eight normal subjects before and after inhalation of six doses of the beta-agonist metaproterenol given over 24 h. After metaproterenol inhalation, beta 2-AR expression as determined by 125I-labeled cyanopindolol binding decreased approximately 70% on bronchial epithelial cells, from 6.3 +/- 0.7 to 2.0 +/- 0.2 fmol/mg (P < 0.001) and to a similar extent on macrophages from 13.3 +/- 0.4 to 3.9 +/- 0.6 fmol/mg (P < 0.001). Agonist inhalation also resulted in impairment of beta 2-AR function in both cell types. With bronchial epithelial cells, maximal isoproterenol-stimulated adenosine 3',5'-cyclic monophosphate (cAMP) accumulation decreased from 9.5 +/- 1.8 to 4.9 +/- 1.2 pmol/10(6) cells (P = 0.003), which amounts to a 48 +/- 6% desensitization. Isoproterenol-stimulated cAMP accumulation in alveolar macrophages decreased from 39.5 +/- 9.0 to 2.9 +/- 0.3 pmol/10(6) cells (P = 0.007), equivalent to 86 +/- 5% desensitization. The cAMP response to forskolin in both cell types was unaffected by metaproterenol inhalation. Thus administration of inhaled beta-agonists results in substantial downregulation and functional desensitization of lung cell beta 2-AR. This supports the concept of a dynamically regulated beta 2-AR in humans, the function of which can be attenuated in relevant target tissues by administration of standard doses of beta-agonist. PMID- 7491993 TI - cAMP generation inhibits inositol 1,4,5-trisphosphate binding in rabbit tracheal smooth muscle. AB - Agonist-induced airway contraction involves the generation and subsequent binding of the phosphoinositide-derived second messenger, inositol 1,4,5-trisphosphate [Ins(1,4,5)P3], to its Ca(2+)-mobilizing intracellular receptor. To the extent that regulatory cross-talk is known to exist between different signal transduction pathways, the present study examined whether activation of the adenosine 3',5'-cyclic monophosphate (cAMP)/protein kinase A (PKA) pathway induces altered binding of Ins(1,4,5)P3 to its receptor in membrane homogenates of rabbit tracheal smooth muscle (TSM). In control TSM, monophasic binding curves provided mean +/- SE values for Ins(1,4,5)P3 receptor density (Bmax) and binding affinity (Kd) amounting to 940 +/- 43 fmol/mg protein and 10.7 +/- 1.2 nM, respectively. Relative to control, binding of [3H]Ins(1,4,5)P3 was significantly reduced in paired TSM separately treated with isoproterenol, forskolin, or dibutyryl-cAMP. Ins(1,4,5)P3 binding was inhibited to a level averaging 60% of control binding by maximal concentrations of each agonist, an effect attributed to a reduction in Ins(1,4,5)P3 binding sites rather than altered ligand affinity. Collectively, these findings demonstrate that activation of the cAMP-dependent signaling pathway is associated with inhibition of Ins(1,4,5)P3 receptor binding and implicate a novel mechanism of action of beta-adrenergic agents in preventing and/or reversing airway contraction. PMID- 7491994 TI - General surgeons in the world of gatekeepers. PMID- 7491995 TI - Edgar J. Poth Memorial Lecture. Surgical aspects of the short-bowel syndrome. AB - BACKGROUND: Surgeons are frequently confronted with patients with the short-bowel syndrome. Important surgical issues are maintaining intestinal continuity, treating complications, and performing procedures to improve intestinal function. METHODS: A comprehensive review of the English language literature and the author's own experience were employed to make recommendations about surgical management of the short-bowel syndrome. CONCLUSION: At the time of initial resection, ostomy formation is often prudent. The decision to restore continuity at a later time should balance anticipated functional outcome against potential complications. Several surgical strategies can be employed at reoperation in these patients to minimize further loss of intestine. Prophylactic cholecystectomy should be considered because of the increased risk of cholelithiasis. Gastric hypersecretion rarely requires operative therapy. Surgical therapy for the short-bowel syndrome includes procedures to slow intestinal transit, optimize intestinal function, and increase intestinal surface area. The choice of operation is influenced by intestinal remnant length and caliber and its function. Only a small proportion of patients are candidates for nontransplant procedures, of which intestinal lengthening is most efficacious. Intestinal transplantation, either alone or combined with the liver, is emerging as the most promising therapy in short-bowel syndrome. PMID- 7491996 TI - Jack A. Barney Resident Research Award. Cardiopulmonary hazards of perihepatic packing for major liver injuries. AB - BACKGROUND: Perihepatic packing has been shown to result in pathologic intra abdominal hypertension. Although now recognized as impairing abdominal organ perfusion, the extent to which perihepatic packing affects cardiopulmonary function has not been elucidated. METHODS: We analyzed a 3-year experience with 11 patients who sustained major hepatic injuries requiring perihepatic packing to control hemorrhage. Pertinent hemodynamic indices consisting of pulmonary capillary wedge pressure (PCWP), cardiac index (CI), oxygen delivery index (DO2), and systemic vascular resistance (SVR), and pulmonary indices consisting of peak airway pressure (PAP), mean airway pressure (MAP), static compliance (CST), and PaO2/FiO2 were measured in the surgical intensive care unit immediately before and after packs were removed. RESULTS: Unpacking resulted in a significant increase in CI (3.1 +/- 0.4 to 4.2 +/- 0.6 L/min/m2), DO2 (539 +/- 41 to 689 +/- 43 mL min/m2), CST (26 +/- 6 to 36 +/- 4 mL/cm H2O), and PaO2/FiO2 (162 +/- 44 to 237 +/- 53 cm H2O), as well as a significant decrease in PAP (47 +/- 9 to 29 +/- 6 cm H2O), MAP (34 +/- 4 to 27 +/- 3 cm H2O), PCWP (21 +/- 4 to 13 +/- 3 mm Hg), and SVR (1,239 +/- 162 to 887 +/- 130 dyne/cm5). CONCLUSIONS: Abdominal compartment syndrome following temporary perihepatic packing can result in significant cardiopulmonary compromise. While perihepatic packing can be an early life-saving procedure, timely alleviation of the secondary syndrome may be critical to the ultimate salvage of patients with marginal cardiopulmonary reserve. PMID- 7491999 TI - Reflux esophagitis in humans is mediated by oxygen-derived free radicals. AB - BACKGROUND: Oxidative stress in reflux esophagitis was investigated before and after antireflux surgery. PATIENTS AND METHODS: Oxidative stress was studied in the distal and proximal esophagus of control patients (without esophagitis, but with other gastrointestinal disorders), of patients with various grades of esophagitis (including Barrett's esophagus), and in patients who had a Nissen fundoplication. Oxidative stress was assessed by chemiluminescence, lipid peroxidation (LP), and by measuring superoxide dismutase (SOD). RESULTS: Chemiluminescence and LP increased with the degree of esophagitis and was highest in patients with Barrett's esophagus; SOD decreased with damage, except in cases of Barrett's esophagus associated with mild esophagitis. Chemiluminescence and LP in reflux patients were higher in the distal than in the proximal esophagus, and SOD was lower, whereas no such difference was found in controls. Findings after Nissen fundoplication were similar to those of controls. CONCLUSIONS: Reflux esophagitis is mediated by free radicals depleting SOD. Barrett's esophagus is a severe form of oxidative damage; in some patients, high SOD levels may prevent severe esophagitis. Antireflux surgery prevents oxidative damage. PMID- 7491997 TI - The challenge of therapy for pancreatitis-related common bile duct stricture. AB - BACKGROUND: Opinions regarding the appropriate clinical management of pancreatitis-related common bile duct (CBD) stricture vary considerably. PATIENTS AND METHODS: Nineteen patients with chronic pancreatitis and proven stricture of their CBD were included in this study. Their mean duct diameter was 16 mm, bilirubin was 8.4 mg/dL, and alkaline phosphatase was 784 mIU/mL. RESULTS: Five patients initially treated with endoscopic biliary stent placement are doing well at a mean follow-up of 7 months with only 1 patient requiring a biliary-enteric bypass. Four patients underwent a pancreaticoduodenectomy and the other 10 patients underwent a biliary-enteric bypass. Mean bilirubin and alkaline phosphatase at 13 months after therapy were 0.9 mg/dL and 144 mIU/mL. CONCLUSION: An endoscopically placed biliary stent will relieve obstruction due to the stricture for several months and allow the inflammatory process to follow its natural course. In patients with long-standing permanent biliary stricture, pancreatitis limited to the head of the pancreas, duodenal obstruction, or suspected pancreatic head carcinoma, pancreaticoduodenectomy is the operation of choice. Biliary-enteric bypass in association with gastric or pancreatic bypasses should be reserved for patients with severe inflammatory changes preventing a safe pancreaticoduodenectomy. PMID- 7491998 TI - Complications of needle catheter jejunostomy in 2,022 consecutive applications. AB - BACKGROUND: We commonly use needle catheter jejunostomy (NCJ) for early enteral feeding in selected patients. Review of our approach was prompted by the suggestion that enteral feeding represents a "stress test" for the bowel and may be associated with a high complication rate. MATERIALS AND METHODS: We reviewed patients with NCJ inserted over the past 16 years by prospective database, chart review, and conference minutes, with emphasis on complications. RESULTS: During the conduct of 28,121 laparotomies, 2,022 NCJs inserted in 1,938 patients (7.2%) resulted in 34 NCJ-related complications in 29 patients (1.5%) The most common complication was premature loss of the catheter from occlusion or dislodgment (n = 15; 0.74%), and the most serious was bowel necrosis (n = 3; 0.15%). CONCLUSIONS: Needle catheter jejunostomy may be inserted and used with a low complication rate. Most complications were preventable through greater attention to detail and better monitoring of physical examination of patients with marginal gut function. PMID- 7492000 TI - Thrombotic thrombocytopenic purpura: indications for and results of splenectomy. AB - BACKGROUND: Thrombotic thrombocytopenic purpura (TTP) is a rare, life-threatening disorder of unknown pathophysiology. The role of splenectomy in the multimodality therapy of TTP is controversial. MATERIALS AND METHODS: All charts of patients with TTP at the University of Utah between 1984 and 1994 were reviewed to evaluate various treatment regimens, and specifically, the impact of splenectomy on morbidity and survival. RESULTS: Of the 15 patients identified, 14 underwent initial treatment with plasmapheresis and steroids. Nine patients were treated with medical therapy only, 6 of whom completely recovered, while 3 patients died. Six patients failed plasmapheresis and underwent splenectomy. There were no operative complications or postoperative deaths. All surgical patients had no active disease at last follow-up. CONCLUSION: Plasmapheresis and steroid administration remain the first-line therapy for TTP. This series documents that splenectomy offers excellent results with minimal morbidity and mortality in patients who do not respond to or who relapse after plasmapheresis. PMID- 7492001 TI - Aspiration pneumonia following surgically placed feeding tubes. AB - BACKGROUND: The enteral route is preferred in surgical patients requiring nutritional support; however, controversy surrounds the choice of location of feeding tube placement. Although jejunostomy has been commonly accepted as superior to gastrostomy for long-term nutritional support because of an assumed lower risk of aspiration pneumonia, recent studies suggest that reevaluation of common practices of surgical tube placement is warranted. PATIENTS AND METHODS: We conducted a retrospective chart review of gastrostomy and jejunostomy procedures from 1986 to 1993. Demographic information and complications related to the procedure were reviewed. Aspiration pneumonia was defined as respiratory symptoms, leukocytosis, and infiltrate on chest radiograph. RESULTS: Sixty-nine gastrostomies and 86 jejunostomies were performed during the study period. Six patients were diagnosed with aspiration pneumonia; 2 cases of which occurred with jejunostomy and 4 cases occurred with gastrostomy (P = not significant). CONCLUSIONS: There was no difference in rates of pulmonary aspiration or other complications between gastrostomy and jejunostomy. We suggest that when a surgically placed feeding tube is required, the determination of appropriate procedure be based on clinical factors such as the technical difficulty of the operation or long-term feeding goals. PMID- 7492002 TI - Predictors of amputation for popliteal artery injuries. AB - BACKGROUND: Popliteal artery injuries continue to result in limb loss. This study identifies risk factors that predict amputation. METHODS: Over a 5-year period, a retrospective chart review was conducted of 80 consecutive patients with 81 popliteal artery injuries. RESULTS: The overall amputation rate was 16.5%. Blunt trauma carried a higher rate of amputation (47%) than penetrating injuries (6.2%); P < 0.0001). Associated fractures had a higher amputation rate, regardless of mechanism (odds ratio +2.7, 95% confidence limits 1.2 to 6.2). Fasciotomy at the time of operation was associated with reduced amputation rate. CONCLUSIONS: Blunt injuries and associated fractures carry an increased risk for amputation. Compartmental pressures should be appropriately monitored postoperatively. Fasciotomy at the time of vascular repair may be considered even without evidence of compartment syndrome. PMID- 7492003 TI - Abdominal wall hernias in patients with abdominal aortic aneurysmal versus aortoiliac occlusive disease. AB - BACKGROUND: This study was undertaken to determine the incidence of ventral incisional hernias (VIHs) and inguinal hernias (IHs) in patients with abdominal aortic aneurysmal (AAA) versus those with aortoiliac occlusive disease (AIOD). PATIENTS AND METHODS: The medical records of 193 patients (128 with AAA and 65 with AIOD) who had undergone elective aortic reconstruction were reviewed to determine the number and location of abdominal wall hernias (AWHs). RESULTS: Forty-one AWHs (28 IHs and 13 VIHs) were detected in patients with AAA compared to 13 (11 IHs and 2 VIHs) in patients with AIOD. There was a significantly greater incidence of VIHs in patients with AAA versus patients with AIOD (10% versus 3%, P < 0.05) and recurrent AWHs (28% versus 19%, P < 0.01), but not of IHs (22% versus 17%). CONCLUSION: Patients with AAA have a higher incidence of VIHs and recurrent AWHs--without a corresponding increase in patient-related risk factors--than patients without aneurysm, suggesting that as yet unidentified etiologic factors may contribute to the development of AWHs in these patients. PMID- 7492004 TI - Improved outcome by identification of high-risk nonocclusive mesenteric ischemia, aggressive reexploration, and delayed anastomosis. AB - BACKGROUND: The factors associated with outcome of patients with nonocclusive mesenteric ischemia are poorly defined. METHODS: Over a 7-year period, 34 consecutive patients with nonocclusive mesenteric ischemia were identified. RESULTS: The mean age of the study patients was 63 years (range 31 to 94); 21 of 34 (62%) were men. The mean delay in diagnosis was 31 hours (range 7 hours to 6 days). Seven of 34 (21%) underwent preoperative visceral arteriography. Two of these 7 required surgery, and both died as a result of intestinal infarction. The remaining 27 had the diagnosis made at celiotomy. Among the 29 who were explored, 16 of 29 (55%) had intestinal infarction. Twenty-one of 29 (72%) had segmental bowel injury whereas 8 of 29 (28%) had massive injury. Among those with segmental infarction, primary anastomosis was performed in 12 of 21 patients (57%); 5 of the 12 (42%) died. Nine of 21 patients (43%) underwent delayed anastomosis; 2 of the 9 (22%) died. No patient with massive injury underwent primary anastomosis. Second-look laparotomy was performed on 22 of 29 (76%). Eleven of those 22 (50%) had a further bowel resection. Overall, 16 of 29 (55%) who underwent surgery for nonocclusive mesenteric ischemia are alive. CONCLUSIONS: Improved survival from nonocclusive mesenteric ischemia is dependent upon the identification of high risk groups, aggressive reexploration, and delayed intestinal anastomosis. PMID- 7492006 TI - Decreasing unplanned extubations in the surgical intensive care unit. AB - BACKGROUND: Unplanned extubations are common, but can be life-threatening. METHODS: We conducted a prospective evaluation of all intubated patients in our surgical intensive care unit to examine the effects of three parameters on the likelihood of accidental extubation. The parameters were the method of endotracheal tube fixation, the use of sedation/paralysis, and the use of hand restraints. During the baseline period, tubes were secured with cloth or velcro ties, sedation was used conservatively, and hand restraints were used routinely. A change in one study parameter was made prior to each period. Thus, in period II, tubes were secured using waterproof tape; in period III, tubes were secured with waterproof tape and sedation/paralysis was used liberally; and in period IV, tubes were secured with waterproof tape and limited use was made of hand restraints. RESULTS: Accidental extubations were significantly less frequent when tubes were secured with waterproof tape (P < 0.0001). No difference was seen when sedation was instituted liberally. Restricted use of hand restraints was associated with significantly increased accidental extubations (P < 0.001). CONCLUSIONS: Our data support the use of water resistant tape to secure endotracheal tubes and the routine use of hand restraints. PMID- 7492005 TI - Is octreotide beneficial following pancreatic injury? AB - BACKGROUND: Pancreatic injury is often associated with multiple complications related to uncontrolled pancreatic exocrine secretion, including pancreatic fistula, pseudocyst, and intra-abdominal abscesses. Somatostatin analogues such as octreotide have been shown to decrease pancreas-related morbidity following major pancreatic resection in patients with pancreatic neoplasms and acute severe pancreatitis. This study was conducted to determine whether or not the administration of octreotide influences the incidence and severity of abdominal complications following pancreatic injury. PATIENTS AND METHODS: Patients with intraoperative diagnosis of pancreatic injury over a 6-year period were studied retrospectively. Specific complications assessed include abdominal abscesses, pseudocyst, pancreatitis, and pancreatic fistula. Statistical analysis of qualitative variables was by chi-square analysis, and analysis of quantitative variables by Student's t-test (P < 0.05). RESULTS: Injury to the pancreas was identified in 96 patients. Sixteen early deaths (< 48 hours) and one late death occurred, for a mortality of 18%, leaving 80 patients as the study population; 21 patients received octreotide and 55 patients did not. Pancreatic fistula occurred in 32 patients (40%). When stratified by pancreatic injury severity, there was no significant difference in complication rates, although patients treated with octreotide had a higher rate of fistula formation (48% versus 40%), longer duration of fistula drainage, and longer hospital stay compared with untreated patients. CONCLUSION: Although adverse patient selection may be a factor in this retrospective survey, the magnitude of observed differences raises concerns regarding the empiric administration of octreotide to such patients pending prospective study. PMID- 7492007 TI - Are corticosteroids salvage therapy for refractory acute respiratory distress syndrome? AB - BACKGROUND: Late acute respiratory distress syndrome (ARDS), characterized by progressive pulmonary interstitial fibroproliferation, is associated with mortality > 80%. Although previous large prospective trials failed to show a benefit of steroids in early ARDS, recent small reports describe improved survival in patients with late ARDS. Recognizing the pathogenetic differences between early and late ARDS, we employed steroid therapy in patients with refractory late ARDS. PATIENTS AND METHODS: Over a 5-year period, we treated 6 patients who were dying of isolated refractory ARDS with methylprednisolone sodium succinate (1 to 2 mg/kg every 6 hours). Ventilatory parameters and lung injury scores were serially recorded, and steroids were weaned based on clinical response. RESULTS: Steroids were instituted after 16 days of advanced mechanical ventilatory support. By day 7 of steroid therapy, there was clinically significant improvement in PaO2/FiO2 ratios (84 to 172) and lung injury scores (3.6 to 2.9); 5 patients (83%) survived. CONCLUSIONS: Steroid therapy appears to be effective in patients with refractory late ARDS. Prospective trials are needed to define the indications, timing of intervention, dose and duration, and precautions of steroid therapy. PMID- 7492008 TI - Carcinoembryonic antigen measurements in the management of esophageal cancer: an indicator of subclinical recurrence. AB - BACKGROUND: Detection of subclinical recurrence after surgical resection of esophageal cancer would allow earlier treatment of recurrent disease and potentially offer a better outcome for rescue therapy. METHODS: The utility of serum carcinoembryonic antigen (CEA) assay was evaluated in the management of patients with esophageal cancer. RESULTS: Serum carcinoembryonic antigen was measured preoperatively in 74 patients. Elevation of the CEA level (> 5 ng/mL) was present in 14 patients (19%). There was no relationship between preoperative CEA elevation and the stage of the tumor or the patients' survival. Eighty-three patients had CEA assay at regular follow-up intervals after resection. Objective evidence of recurrent disease was determined at similar intervals by chest radiography and abdominal and thoracic computed tomography scans. During follow up, 53 of 83 patients developed recurrence. Postoperative elevation of CEA levels occurred in 32 patients, resulting in a sensitivity of 55% for detecting recurrent disease. Twenty-nine of the 32 patients who developed CEA elevation had objective evidence of metastatic disease. In 13 patients, the rise in CEA levels predated objective evidence of recurrence by a median of 4 months (range 3 to 35), and in 16 patients, it occurred concomitantly. The specificity with which an elevated postoperative CEA level indicated recurrence was high, 90%, with a positive predictive value of 91%. CONCLUSIONS: Postoperative CEA elevation is highly predictive of recurrent disease. In 16% of patients, elevation of CEA was the earliest objective sign of recurrence; such elevation should prompt consideration of adjuvant therapy. PMID- 7492010 TI - A half century of experience with carcinoid tumors in children. AB - PURPOSE: To investigate the frequency, presentation, clinical management, and prognosis of appendiceal carcinoid tumors in children. METHOD: A review of our institution's experience over 50 years. RESULTS: Twenty-two patients below the age of 20 presented with appendiceal carcinoid tumor. The mean age at presentation was 14.6 years. Twelve patients presented with symptoms of appendicitis. No tumor was > 2.0 cm in size. Only 2 patients underwent resection beyond appendectomy. No patient had recurrent or metastatic carcinoid tumor, and all but 1 patient (who died of ovarian choriocarcinoma) are alive without evidence of carcinoid tumors 1.5 to 30 years after diagnosis. CONCLUSIONS: Appendiceal carcinoid tumors in children are rarely life-threatening and the incidence of large tumors (> 2.0 cm) is very low. The role of right hemicolectomy in large (> 2.0 cm) tumors is questionable in this age group. PMID- 7492009 TI - A selective therapeutic approach to gastric cancer in a large public hospital. AB - BACKGROUND: Gastric cancer is a common malignancy with a poor prognosis. The improved survival reported from Japan may be due to earlier detection or to more radical surgery, or both. The relevance of their methods to gastric cancer seen in Western countries is uncertain. PATIENTS AND METHODS: The study involved 204 patients with gastric carcinoma. Preoperative staging by computed tomography scan and endoscopic ultrasound showed that 120 patients (59%) had stage IV disease. RESULTS: Curative resection was performed in 66 patients, palliative resection in 32, bypass/intubation in 39, chemotherapy alone in 41, and supportive treatment in 26. Neoadjuvant chemotherapy was given to 40 of 66 patients treated with curative resection. The mortality of gastrectomy was 3%. Survival was significantly improved after curative resection compared with palliative resection, which in turn was improved over non-resectional or nonsurgical therapy. Postoperative morbidity included four intra-abdominal abscesses, all associated with splenectomy. CONCLUSIONS: Curative surgery for gastric cancer is worthwhile, but the advanced stage of the disease in a public hospital should encourage the establishment of a screening program in high risk populations. PMID- 7492011 TI - Preoperative chemoradiation downstages locally advanced ultrasound-staged rectal cancer. AB - BACKGROUND: This prospective study assessed the effect of preoperative radiation and chemotherapy on the pathologic staging of advanced rectal cancer. METHODS: Twenty patients with rectal cancer were treated with combined chemoradiation prior to operation, after pretreatment staging of all lesions with transrectal ultrasound (TRUS). Perirectal fat invasion served as minimal criteria for preoperative neoadjuvant therapy. The pretreatment stage of these rectal lesions as defined by TRUS was then compared with the pathological stage of the surgical specimen following resection. Cancers were treated with high-dose radiation (45 to 54 Gy) in 19 of 20 patients. One patient received in excess of 60 Gy because of tumor characteristics. Chemotherapy consisted of 5-fluorouracil delivered as a continuous infusion or bolus therapy. Four to 8 weeks after neoadjuvant therapy, 13 abdominal perineal resections, 5 low anterior resections, and 2 completion proctectomies were performed. RESULTS: Following resection, rectal cancer was downstaged in 14 of 20 patients. No tumor was present in the rectal wall in 8 of 20 patients. Complete pathological response was present in 7 of 20 patients. Local recurrence occurred in 2 of 20 patients. Disease-free survival in the remaining 17 of 20 patients ranges from 9 to 51 months (average 26). CONCLUSIONS: Preoperative chemoradiation in the surgical management of advanced rectal cancer results in demonstrable tumor downstaging. PMID- 7492012 TI - Minimally invasive surgery for gastroesophageal reflux disease. AB - BACKGROUND: The goal of this study was to determine if the outcome of antireflux surgery can be improved by: (1) conducting a careful preoperative workup to characterize gastroesophageal reflux disease (GERD) in the individual patient; and (2) tailoring the operation to the results of the preoperative function tests. PATIENTS AND METHODS: Sixty-eight patients had operations for GERD by minimally invasive techniques. RESULTS: A Rossetti fundoplication was performed in 22 patients. Sixty-eight percent became asymptomatic. Twenty-seven percent developed dysphagia or gas bloat. Thirty-five patients had a Nissen fundoplication. Ninety-one percent are asymptomatic. Eleven patients with severe abnormalities of esophageal peristalsis underwent a Guarner fundoplication with relief of symptoms in 82% of patients. No patients in the Nissen or Guarner group developed postoperative persistent dysphagia or gas bloat. A pyloromyotomy was performed in 3 patients because of severe delayed gastric emptying. CONCLUSIONS: Minimally invasive surgery for GERD gives good-to-excellent results even in patients with abnormal esophageal body function, provided that the operation is tailored to the individual patient based on the results of the preoperative function tests. PMID- 7492013 TI - Managing choledocholithiasis in the laparoscopic era. AB - BACKGROUND: Management options for common bile duct stones have explained in the era of laparoscopic cholecystectomy (LC), and selecting the most appropriate method for each patient can be problematic due to the difficulty of predicting accurately which patients have choledocholithiasis (CDL). In order to improve selection of appropriate treatment for CDL, treatment options were analyzed for outcome retrospectively during a 25-month period beginning June 1, 1992. PATIENTS AND METHODS: Four hundred four patients underwent LC; 48 (12%) had CDL identified at preoperative endoscopic retrograde cholangiopancreatography (ERCP) or intraoperative cholangiogram (IOCG). Forty-seven patients were referred for preoperative ERCP for suspected CDL, 23 (49%) of whom had proven duct stones and underwent endoscopic sphincterotomy and stone retrieval with an 87% success rate. RESULTS: Of 357 patients scheduled for LC without preoperative ERCP, 236 had IOCG, of which 25 (11%) demonstrated CDL. Seven patients had open common bile duct exploration (CBDE). Sixteen patients had postoperative ERCP after positive IOCG, 7 (44%) of which were positive for CDL and whose stones were removed with 100% success. Two patients were observed, anticipating spontaneous passage of a small stone. CONCLUSIONS: Preoperative ERCP should be applied selectively. For the large majority of patients without preoperative evidence of CDL, we recommend routine IOCG; if CDL is demonstrated, an intraoperative decision can be made to proceed to postoperative ERCP in the usual case or to open CBDE for very large or multiple stones. Observation in anticipation of spontaneous passage may be appropriate for small, solitary common duct stones. Continuing advances in laparoscopic CBDE are likely to reduce further the need to rely on ERCP in managing CDL. PMID- 7492014 TI - Laparoscopic splenectomy offers advantages in selected patients with immune thrombocytopenic purpura. AB - BACKGROUND: Splenectomy plays a crucial role in the management of selected patients with immune thrombocytopenic purpura (ITP). Laparoscopic splenectomy has been suggested as an alternative to open techniques. MATERIALS AND METHODS: A retrospective chart review was made for all patients undergoing splenectomy for ITP at our institution from June 1987 to August 1994. RESULTS: Twenty-one patients underwent splenectomy for ITP. Fourteen open procedures (group 1) and 7 laparoscopic procedures (group 2) were performed. Eleven of 14 patients in group 1 and all 7 patients in group 2 had appropriate increases in platelet count following surgery. There were 3 complications in group 1 and no major complications in group 2. Operative time was longer in group 2 (154 versus 68 minutes, P < 0.05), but decreased with experience (210 to 135 minutes). Compared with patients in group 1, patients in group 2 tolerated liquids sooner (0.7 versus 2.6 days, P < 0.05), required less parenteral narcotic (14 versus 65 morphine equivalents, P < 0.05), and were discharged sooner (2.1 versus 5 days, P < 0.05). All patients in group 2 remain well and require no medication after a mean follow-up of 11 months. CONCLUSIONS: Laparoscopic splenectomy is safe and effective in the management of select patients with ITP and offers advantages over open techniques. PMID- 7492015 TI - Evaluation of video-assisted thoracoscopic surgery in the diagnosis of diaphragmatic injuries. AB - BACKGROUND: Injury to the diaphragm from penetrating or blunt thoracoabdominal trauma is notoriously difficult to diagnose. Chest radiography, computed tomography scan, contrast studies, diagnostic peritoneal lavage, and laparoscopy are inadequate; thus, celiotomy is commonly performed in patients with suspected diaphragmatic injury. We compared the diagnostic accuracy of video-assisted thoracoscopic surgery (VATS) with that of exploratory celiotomy in the evaluation of diaphragmatic and thoracoabdominal injury. PATIENTS AND METHODS: Hemodynamically stable patients admitted to a level I trauma center with blunt or penetrating injury to the lower chest or abdomen underwent VATS and subsequent celiotomy under the same general anesthetic. Intraoperative thoracoscopic findings were blinded to the abdominal surgeons. RESULTS: Twenty-six patients were enrolled in the study over a 12-month period. Diaphragmatic injuries were identified in 8 patients (31%). Videothoracoscopy identified all eight injuries in these patients. Six of the 8 patients (75%) with diaphragmatic injuries sustained associated injury to intrathoracic or intra-abdominal organs. There was no mortality and no procedure-related morbidity. There were no missed injuries in patients who underwent VATS. CONCLUSIONS: Video-assisted thoracoscopy is a safe, expeditious, and accurate method of evaluating the diaphragm in injured patients, and is comparable in diagnostic accuracy to exploratory celiotomy. PMID- 7492017 TI - Credibility and clinical utility of thyroid fine-needle aspiration biopsy in a teaching hospital. AB - BACKGROUND: Fine-needle aspiration (FNA) biopsy allows cost-effective thyroid nodule management at referral centers. The authors examined its use at a teaching hospital for safety, accuracy, and clinical utility. METHODS: Patients having thyroid FNA biopsy in 1991 were identified. Pathology results through 1993 were retrieved and selected charts through 1993 were reviewed. House staff, with faculty supervision, performed patient evaluations, FNA biopsy, and therapy. RESULTS: There were 91 uncomplicated FNAs. Aspirates were diagnostically adequate in 78 (86%) cases. In 15 operations for neoplastic FNA biopsy, 14 (93%) tumors were confirmed. Twelve operations for clinical indications despite benign FNA biopsy yielded 2 (17%) adenomas. Fine-needle aspiration biopsy and frozen section were similarly accurate. No observed patients have had thyroidectomies. Patient management, operation in 15 cases and observation in 48 cases, was directed by FNA biopsy in 63 of 91 patients (69%). CONCLUSION: Thyroid FNA biopsy by house staff with faculty supervision is safe, accurate, allows efficient patient management, and can reduce frozen section use. PMID- 7492016 TI - Therapeutic laparoscopy in trauma. AB - PURPOSE: To assess the therapeutic potential of emergent laparoscopy in the trauma setting, a retrospective review was performed in a busy urban trauma center. PATIENTS AND METHODS: Between December 1991 and October 1993, 133 hemodynamically stable patients with suspected abdominal injury were evaluated laparoscopically. All laparoscopic procedures were performed in the operating room under general anesthesia. Mechanism of injury was stab wound (58), gunshot wound (57), and blunt trauma (18). No significant injuries were found in 72 patients (54%), and these patients received no further treatment. On the basis of laparoscopic findings, 52 patients underwent formal exploratory laparotomy. Surgical exploration confirmed the presence of significant injuries in 44 of the 52 patients (85%). Therapeutic laparoscopy was performed in 6 patients (5%) for diaphragm repair (4), gastrotomy repair (1), and splenorrhaphy (1). Additionally, 10 patients underwent laparoscopy-guided blood salvage for autotransfusion during laparoscopic evaluation of blunt trauma. Three small-bowel enterotomies were repaired during minilaparotomy. RESULTS: No significant injuries were missed as a result of our use of laparoscopy in trauma assessment. Complications--trocar enterotomy, trocar laceration of the inferior epigastric artery, and transient hypotension--occurred in 3 patients secondary to the use of laparoscopy. CONCLUSIONS: Trauma laparoscopy is a safe method for the evaluation of selected patients with abdominal trauma and can reduce the number of negative and nontherapeutic trauma laparotomies performed. Limited therapeutic intervention is possible in a small number of patients. PMID- 7492018 TI - Surgical margins after needle-localization breast biopsy. AB - BACKGROUND: The use of needle-localization breast biopsy (NLBB) for the early diagnosis of breast cancer is common. The therapeutic adequacy of tumor-free margins following NLBB is unknown. We hypothesized that the presence of residual tumor after reexcision (mastectomy, tylectomy, or quadrantectomy) does not depend on the margin status following NLBB. PATIENTS AND METHODS: Retrospective cohort analysis was performed on 890 consecutive NLBBs executed between January 1990 and June 1994. Patients with invasive breast neoplasia were divided into two groups based on the tumor margins after NLBB. Group 1 were the women with positive margins, and group 2 had negative margins. Breast specimens after reexcision were reviewed for evidence of residual invasive carcinoma. RESULTS: Invasive neoplasia was present in 107 patients (12%). Surgical margins and definitive records of care were available for 96 of them (90%). All 45 patients in group 1 and 38 (75%) of 51 patients in group 2 underwent reexcision of the initial biopsy site (P = 0.36). Residual invasive carcinoma was present in 10 patients (22%) in group 1 and 3 (8%) in group 2 (P = 0.13). CONCLUSION: Invasive breast neoplasia diagnosed by NLBB requires reexcision regardless of tumor margins to achieve complete local surgical eradication of tumor. PMID- 7492019 TI - Modified ilioinguinal node dissection for metastatic melanoma. AB - BACKGROUND: Standard ilioinguinal node dissection for melanoma has substantial cost and morbidity. Beginning in 1988, we modified the procedure in hopes of reducing side effects without compromising survival. PATIENTS AND METHODS: Dissection was standard except for preservation of saphenous vein and femoral sheath and omission of sartorius muscle transfer. To date, 19 patients with recurrent melanoma in the groin have had the procedure, 6 for N1 disease and 13 for N2, M1 metastases. RESULTS: Average hospital stay was 4.5 days (range 3 to 7). Postoperative edema occurred in 1 (5%) patient. Disease-free survival at 40 months was 66% for N1 disease and 26% for N2, M1 metastases. CONCLUSION: Modified ilioinguinal node dissection appears to reduce cost and morbidity of treating recurrent melanoma in the groin without compromising survival. PMID- 7492021 TI - Problem orientation is a new approach to surgical education. AB - BACKGROUND: There is increasing interest in educational methods that are loosely aggregated under the title of problem-based learning (PBL), but it remains unclear whether PBL is as successful as its conventional predecessor in transmitting factual information. MATERIALS AND METHODS: The authors designed and implemented a PBL curriculum for a third-year surgical clerkship, then prospectively compared that technique with the conventional format. Each student's subject-related knowledge was assessed with a specifically tailored 195 question written exam and correlated with National Board of Medical Examiners shelf exams. Student and faculty responses to the technique were also sought and tabulated. RESULTS: Student and faculty responses to PBL were uniformly positive. We were unable, however, to demonstrate effects on our evaluation instruments. Neither individual student performance nor grouped scores differed based on the mode of presentation. CONCLUSION: A PBL curriculum generates both student and faculty enthusiasm. Unfortunately, this does not translate into more efficient transmission of knowledge. PMID- 7492020 TI - Is academic managed care an oxymoron? AB - BACKGROUND: A review of 1993 data on length of stay (LOS) and charges for diagnosis-related group (DRG) 195 (complicated cholecystectomies) showed that Maricopa Medical Center charged more and had longer LOS than all other area hospitals. METHODS: Twenty DRG 195 charts were analyzed for the causes of the inefficiencies. The remaining cholecystectomy DRGs were similarly analyzed. RESULTS: Analysis of the charts for DRG 195 showed that 55% of the patients had laparoscopic conversions. Charges and LOS varied significantly because of the conversions, increased preoperative hospital days and increased operative times. Moreover, 30% of patients were more than 70 years old. Comparisons of other cholecystectomy DRGs showed similar inefficiencies, indicating a hospital system's problem. CONCLUSIONS: Diagnosis-related group delta analysis is a powerful performance improvement tool. Once problem areas are identified and corrected, monitoring prospective data produces rapid analysis of quality of care and cost improvements. The models can serve as a means for teaching hospitals to become more competitive and satisfy the Joint Commission on Accreditation of Healthcare Organizations requirements for patient-care improvements. PMID- 7492022 TI - Risks, costs, and the expected complication of re-injury. AB - BACKGROUND: The purpose of this study was to determine the characteristics of young victims of violence and the risk of rehospitalization due to intentional injury (recidivism) and to estimate the potential cost of these injuries. MATERIALS AND METHODS: Trauma admissions from January 1, 1991 to December 31, 1993, at San Francisco General Hospital of youths < 25 years old who were victims of gunshot wounds (GSWs), assault, and stab wounds were screened. Five hundred and fifty-two charts were reviewed after sampling every other chart. The cost of hospitalization was estimated from the Medicare charge-to-cost ratio. RESULTS: There were 87 (16%) persons who had a prior injury, of whom 82 (94%) had suffered their injury within the previous 5 years. The predominate mechanism of injury was GSW (242, 44%). There were 38 deaths; 35 (92%) were by firearms. The estimated cost of hospitalization for 552 youths for 3 years was $3,843,545.58. CONCLUSIONS: Intentional injury is a major risk factor and potential predictor for re-injury. Firearms are a major mechanism of intentional injury among youths and a major determinate of death. With the estimated cost of $2,562,363.72 per year for all youths at our hospital, intentional injury is a major health care issue for youths and hospitals. PMID- 7492023 TI - Comparison of diagnostic specimens and methods to evaluate infected venous access ports. AB - BACKGROUND: Implanted venous access port infection can be difficult to diagnose and treat. If device removal is necessary, confirming port infection is problematic. MATERIALS AND METHODS: Culture specimens from three sites, catheter tip (Tip), port pocket, and the material within the reservoir (Inside), were sent from ports removed for potential infection. The results of these cultures were compared to preremoval peripheral and central blood cultures. RESULTS: Forty-five ports were removed for suspected infection. Confirmed port infection was defined as positive culture(s) from one or more experimental specimen(s). In 29 evaluable cases, the Inside specimens were completely predictive. Tip specimens were less accurate, even with a lower diagnostic threshold. In 7 of 19 confirmed infections, only the Inside culture was diagnostic. CONCLUSION: The most predictive culture specimen in a potentially infected port is the thrombotic material inside the reservoir. PMID- 7492024 TI - Orthotopic liver transplantation with selective use of venovenous bypass. AB - BACKGROUND: To determine the utility of selective use of venovenous bypass (VVB), an algorithm based upon hemodynamic criteria was instituted at Stanford University Medical Center: the bypass was used if the systolic blood pressure decreased below 100 mm Hg with a trial of caval and portal clamping. PATIENTS AND METHODS: Eleven consecutive patients underwent orthotopic liver transplantation (OLT) with use of VVB on a selective basis; using the hemodynamic exclusion criteria, none required VVB. A group of 20 patients undergoing OLT with VVB served as historical controls. RESULTS: Overall patient and graft survival were identical in both groups (75%). Avoidance of VVB decreased operative and warm ischemia time and decreased peak transaminase and total bilirubin values, but increased rates of intraoperative blood loss. However, the absolute numbers of blood products administered were not different between groups. CONCLUSION: Selective use of VVB for OLT does not incur increased morbidity or mortality. Potential advantages include cost savings with decreased operative and anesthetic time. PMID- 7492025 TI - The safety of orotracheal intubation in patients with unstable cervical spine fracture or high spinal cord injury. AB - BACKGROUND: The potential merits and dangers of orotracheal and nasotracheal intubation in patients with injury to the cervical spine or spinal cord continue to be debated. To address this issue, a prospective study was conducted at a level 1 trauma center in patients with respiratory embarrassment and either or both of these injuries. MATERIALS AND METHODS: Over a 7-year period, all such patients underwent neurologic examination by a trauma surgeon on arrival at the trauma center, immediately after endotracheal intubation, and at frequent intervals throughout hospitalization. Cervical immobilization was maintained manually during endotracheal intubation. When necessary, patients were sedated or paralyzed with short-acting pharmacologic agents. RESULTS: During the study period, there were 81 patients with 98 cervical vertebral body fractures, but without evidence of spinal cord injury on initial examination. Sixty-seven patients (83%) were legally intoxicated, and 12 patients had closed head injury. Endotracheal intubation was performed in 26 patients with unstable fractures, and 22 patients were intubated via the oral route. No patient manifested a subsequent neurologic deficit. Sixty-nine additional patients presented with high spinal cord injury; 16 had no cervical spine fracture, and 53 patients had 61 fractures of the cervical vertebrae. Sixty patients (87%) were intoxicated, and 8 patients had closed head injury. Endotracheal intubation was performed in 29 of these patients, and 26 patients were intubated via the oral route. No patient experienced further neurologic deficit following endotracheal intubation. CONCLUSION: In trauma victims with or at high risk of cervical spinal cord injury, orotracheal intubation is a rapid, safe means of achieving airway control. PMID- 7492026 TI - Evaluation of the thoracic and lumbar spine after blunt trauma. AB - BACKGROUND: Routine performance of thoraco-lumbar (TL) spinal radiology in patients with blunt trauma is controversial. PATIENTS AND METHODS: To establish indications for radiologic screening of the TL spine, a retrospective review of 344 patients who had radiologic evaluation of the spine was performed. RESULTS: Forty-seven patients had abnormalities detected on radiologic evaluation. One hundred eighty-six patients had at least one clinical finding suggestive of injury. Thirty-two had abnormal radiographs. Thirteen of these abnormalities represented old or minor fractures and were not treated. Nineteen patients had injuries requiring treatment. Two patients were treated with activity restriction, 12 with a back brace, 4 with operative fixation, and 1 patient died prior to operation. Of the 129 patients who were awake, alert, and without clinical evidence of injury, 10 had abnormal radiologic studies. Three patients had old fractures, 4 had transverse process fractures, and 3 had spondylolisthesis. None of these patients required treatment. Twenty-nine patients had equivocal clinical examinations primarily due to altered levels of consciousness. Five patients had abnormal radiologic studies, 3 of whom required treatment. Three factors associated with the occurrence of TL spine injury were identified: an Injury Severity Score > or = 15, a positive clinical examination, and a fall of > or = 10 feet. CONCLUSIONS: These data suggest that patients who are awake, alert, and with no clinical evidence of injury do not require radiologic study of the TL spine. Patients with equivocal or positive clinical findings or with altered levels of consciousness should have complete TL spine evaluation. PMID- 7492027 TI - Changing trends in the management of splenic injury. AB - BACKGROUND: A gradual change in the management of splenic injuries has occurred at our institution. This study was therefore undertaken to determine whether changes in management of splenic injury influenced outcomes during the past 30 years. PATIENTS AND METHODS: A retrospective study of patients admitted with splenic trauma between 1965 and 1994 was performed. Two hundred seven patients were identified and demographic and outcome data were recorded. Patients were then grouped based upon the period in which they received treatment (ie, Period I [1965 to 1974], Period II [1975 to 1984], and Period III [1985 to 1994]), and the type of treatment received (ie, splenectomy, splenorrhaphy, or observation). RESULTS: More patients were treated in Period III than in the other two periods, and Period III patients had shorter hospital stays. Splenectomy was solely used during Period I; splenorrhaphy and observation were occasionally performed during Period II; and splenectomy, splenorrhaphy, and observation were performed in near equal numbers during Period III. Mortality was similar for each period, though Injury Severity Scores (ISS) were higher during later years. When compared by treatment modality, patients receiving splenectomy had higher ISS and splenic injury classifications. CONCLUSION: Patients treated by splenorrhaphy and observation for splenic injury have markedly increased over the past 30 years without adverse outcome. PMID- 7492031 TI - Unilateral breast masses in men over 40: a diagnostic dilemma. PMID- 7492030 TI - Does level of ligation influence results in a murine biliary obstruction model? PMID- 7492028 TI - Severe acidosis alone does not predict mortality in the trauma patient. AB - BACKGROUND: Because severe acidosis is an indicator of poor prognosis in trauma patients, medical records of these patients were analyzed to determine whether aggressive resuscitation was appropriate. PATIENTS AND METHODS: Data from a level 1 trauma center registry were reviewed retrospectively to identify patients with a pH < or = 7.0. Thirty-seven patients were identified. Severely acidotic patients were compared to average trauma patients in terms of demographics, resuscitation, injury, and outcome. Surviving acidotic patients were also compared to nonsurviving acidotic patients. RESULTS: Half of the severely acidotic group survived initial resuscitation with approximately one third surviving to leave the hospital. There were no chronically disabled survivors. Nonsurviving acidotic patients were more unstable, more neurologically depressed, and more severely injured. Resuscitation efforts did not consume excessive hospital resources. CONCLUSION: Severe acidosis alone is not a sufficiently powerful predictor of outcome to justify withholding resuscitation; however, when combined with coma and shock, this condition had no survivors in this small series. PMID- 7492029 TI - Survival in major burn injuries is predicted by early response to Swan-Ganz guided resuscitation. AB - BACKGROUND: Two years ago the authors began to use Swan-Ganz catheters to generate more complex hemodynamic data as a better guide to resuscitation of burns. This study uses the information to identify differences between survivors and nonsurvivors. PATIENTS AND METHODS: Fifty-three consecutive patients with major burns were treated using Swan-Ganz-generated data. Additional information was collected, including demographics, intake and output, medications, and arterial blood gas (including calculated oxygen consumption). Statistical analysis incorporated adjustments for autocorrelation. RESULTS: The patients included 38 men and 15 women, averaged 43.7 years of age, and had a mean burn size of 40%. Thirteen patients suffered severe inhalation injury. Data collected hourly over 3 days were collapsed across 6-hour observation periods. Mean values, when plotted across time, discriminated the 37 survivors from the 16 nonsurvivors. Poor response to resuscitation as evidenced by increased use of colloid fluid and cardiotonic drugs plus failure to maximize oxygen consumption were associated with nonsurvival. CONCLUSIONS: Unsustained or inadequate response to hyperdynamic resuscitation of burns was associated with nonsurvival. PMID- 7492033 TI - Validation of a modified spectrophotometric method for the determination of nitrate in dry milk using 2-sec-butylphenol. AB - A rapid, simple and direct visible spectrophotometric method, based on the quantitative nitration of 2-sec-butylphenol in concentrated sulfuric acid, is described for the determination of nitrate in milk. The nitration product was extracted into toluene and the yellow nitrophenoxide compound formed in alkaline aqueous solution was determined at 418 nm. Carrez reagents were used for protein precipitation. Interference from lactose, chloride and nitrite ions and the effect of reagent concentration was controlled. The calibration graph was linear over the range 0.5-5 micrograms ml-1 NO3-; y = -0.0019 + 0.0681 x; r = 0.9997. The limits of detection and quantification were found to be 0.18 and 0.5 microgram ml-1 NO3-, respectively. The accuracy of the method did not depend on nitrate content in spiked milk. The mean recovery of the spiked milk was found to be 99.81% with the confidence limits of 0.67%, a relative standard deviation (RSDr) of 1.79% for repeatability and an RSDR value of 1.90% for reproducibility. Adequate agreement was found between results obtained by the nitration method and those of the French official reduction spectrophotometric reference method (AFNor). For the Indonesian dry (powdered) milk sample studied the nitrate levels were in the range 10.7-29.5 mg kg-1 NO3-. PMID- 7492034 TI - Chest pain associated with cocaine: an assessment of prevalence in suburban and urban emergency departments. AB - STUDY OBJECTIVE: Chest pain and myocardial infarction following the use of cocaine have been well documented. We assessed the prevalence of cocaine use in patients who presented to the emergency department with chest pain of possibly ischemic origin. DESIGN: During times of research assistant availability, consecutive adults with the chief complaint of chest pain unexplained by trauma or radiographic abnormality were questioned about cocaine use in the preceding week. Urine was tested for the presence of cocaine or cocaine metabolites with a highly accurate bedside urine test kit (specificity, 100%; sensitivity 98%). Anonymous unlinked data-collection methods were used. Therefore we could not determine whether the patients who used cocaine had sustained myocardial infarctions. SETTING: One suburban and three urban EDs. RESULTS: We enrolled 359 patients with a mean age of 51 years, 8% of whom sustained myocardial infarctions. Sixty patients (17%) had cocaine or cocaine metabolites in urine. The likelihood of testing positive for cocaine varied by age group: 18 to 30 years, 29%; 31 to 40 years, 48%; 41 to 50 years, 18%; 51 to 60 years, 3%; 61 years or older, 0% (P < .0001). Of the 60 patients who tested positive for cocaine, only 43 (72%) admitted recent use. CONCLUSION: Many ED patients with chest pain have recently used cocaine. Because the recent use of cocaine is not uncommon in patients with chest pain up to 60 years old, such patients should be questioned about cocaine use. When treatment or disposition may be altered, consideration should be given to objective assessment of cocaine use because patient self-report does not appear reliable. PMID- 7492032 TI - Splenic preservation based on preserved histology and phagocytic function using upper short gastric vessels. PMID- 7492035 TI - Randomized double-blind controlled trial comparing room-temperature and heated lidocaine for digital nerve block. AB - STUDY OBJECTIVE: To determine whether warming of lidocaine decreases the pain of its injection during digital nerve block. DESIGN: Prospective, randomized, double blind, controlled trial. INTERVENTIONS: Twenty healthy volunteers received bilateral digital nerve blocks of their middle finger. They were first randomly assigned to receive either room-temperature (21 degrees C) or heated (42 degrees C) 2% lidocaine in their first block. They were then randomly assigned to receive the first block in either the right or left hand. The blocks were performed in a standardized fashion by a single physician, who was blinded to which solution was being used. The volunteers rated the pain of each digital block on a 100-mm visual analog scale (VAS). Efficacy of each digital block was tested at 5 minutes. RESULTS: Heating of the lidocaine was associated with a significantly lower median VAS pain score (31.5 versus 25.0; P < .05). There was no difference in pain score between the two solutions in relation to which hand was used (P = .29) or whether the injection was the first or the second (P = .37). When all factors (temperature, order, and hand) were considered in the ANOVA with respect to VAS pain score, the only significant relation found was that between the temperature of the solution and the VAS pain score (P = .028). CONCLUSION: Heating of lidocaine decreases the pain of injection during digital nerve block. PMID- 7492038 TI - Effect of adenosine on the management of supraventricular tachycardia by urban paramedics. AB - STUDY OBJECTIVE: To determine the effect of the addition of adenosine, as a standing-order medication, on the prehospital management of supraventricular tachycardia (SVT) in a large urban emergency medical services (EMS) system. DESIGN: Prospective observational case series with historical controls. SETTING: Large urban municipal EMS system staffed by paramedics and emergency medical technicians trained to operate automatic or semiautomatic defibrillators (EMT Ds). PARTICIPANTS: We observed a consecutive sample of prehospital patients who presented with an initial ECG rhythm of SVT, as interpreted by the treating paramedics, between July 1 and December 31, 1993. We used patients from the same 6-month period in 1992 as our control group. Indications for treatment were chest pain, evidence of myocardial ischemia, or shock. Adenosine had been introduced as a first-line medication to be used under standing orders in cases of unstable SVT before a physician was contacted for medical control options. RESULTS: We studied 239 cases and 228 controls. Acceptable call reports with pretreatment and posttreatment ECGs were available for 140 (59%) of the study cases and 104 (46%) of the controls. The two groups were similar in terms of age, sex, and initial vital signs. In the control group, 75 patients had indications for treatment, and 16 were treated (21%). In the study group, 127 had indications for treatment and 103 (81.1%) were treated (odds ratio, 15.83; 95% confidence interval, 7.38-34.4). CONCLUSION: The introduction of adenosine as a standing-order medication into an urban EMS system increased the proportion of patients who received advanced life support treatment. Paramedics were able to accurately diagnose and begin treatment of SVT with adenosine without direct medical supervision. PMID- 7492037 TI - Using the "GI cocktail": a descriptive study. AB - STUDY OBJECTIVE: To determine practice patterns regarding administration of the "GI cocktail" (a mixture of liquid antacid, viscous lidocaine, and an anticholinergic) in the emergency department in a single hospital and the responses and final dispositions of patients who received the cocktails. DESIGN: A retrospective review of ED charts. SETTING: Urban university hospital ED with an annual census of 50,000 visits. PARTICIPANTS: Ninety-seven consecutive patients who received a GI cocktail in the ED. RESULTS: Forty-nine patients (50%) received a GI cocktail for a chief complaint of abdominal pain and 40 (41%) for a chief complaint of chest pain. The reason for administration of a GI cocktail was documented on only one chart. Sixty-six patients (68%) received at least one other drug, at a median time of 9 minutes before administration of the GI cocktail. The most commonly coadministered drug was a narcotic (56 patients), followed by nitroglycerin (22 patients), antiemetics (13 patients), H2-blockers (13 patients), and aspirin (10 patients). Thirty-three patients (34%) had symptomatic relief with the cocktail alone, 35 (36%) had symptomatic relief with the cocktail plus other drugs, 7 (7%) had no response to the GI cocktail alone, and 5 (5%) had no response to the cocktail with other drugs. In 17 patients (18%) the response was not documented. Chest pain patients and abdominal pain patients had a similar frequency of response. There was also similarity of response between admitted and discharged patients. CONCLUSION: Although symptomatic relief after administration of a GI cocktail is often noted, it is difficult to differentiate the effects of the cocktail from those of other coadministered medications such as morphine or nitroglycerin. We urge ED physicians to use the GI cocktail in a rational manner. PMID- 7492036 TI - Randomized double-blind comparison of the analgesic efficacy of intramuscular ketorolac and oral indomethacin in the treatment of acute gouty arthritis. AB - STUDY OBJECTIVE: To compare the analgesic effect of IM ketorolac tromethamine with that of oral indomethacin in the treatment of acute gouty arthritis. DESIGN: Prospective, randomized, double-blind, controlled, parallel group clinical trial. SETTING: Two urban emergency departments. PARTICIPANTS: Twenty consecutive patients who presented to the ED with acute gout. INTERVENTIONS: Each patient was randomly assigned to receive in the ED (1) 60 mg of IM ketorolac and oral placebo or (2) 50 mg of oral indomethacin and IM placebo. The patients rated the intensity of their pain on a Wong-Baker pain scale (which runs from 0 to 5) before treatment and 30, 60, 90, and 120 minutes after treatment. All the patients were discharged with instructions to take oral indomethacin and to complete pain score cards at home at 6, 12, and 24 hours. RESULTS: The 10 patients in each group were similar with regard to age, sex, race, and initial mean pain score. After 2 hours, the mean pain scores (+/- SD) for the ketorolac group had decreased from 4.5 +/- .71 to 1.4 +/- 1.43 (P < .05), and the mean score for the indomethacin group had decreased from 4.4 +/- .70 to 1.5 +/- 1.18 (P < .05). The difference between the two groups was not significant. At 6 hours, there was some pain rebound in the ketorolac group. CONCLUSION: IM ketorolac and oral indomethacin are similar in the relief of the pain of acute gouty arthritis in the ED. PMID- 7492039 TI - Measuring the call-receipt-to-defibrillation interval: evaluation of prehospital methods. AB - STUDY OBJECTIVE: Successful resuscitation of cardiac arrest depends partly on the time of first defibrillation. An accurate, practical method of measuring this time has not been devised. We attempted to determine the interval from receipt of a call by emergency medical services personnel to first defibrillation (total defibrillation interval) with synchronized clocks between computer-aided dispatch operations and an event-recording defibrillator. DESIGN: A 7-month prospective study measuring the total defibrillation interval. An automated code summary was to be submitted for each participant. SETTING: An urban, all-advanced life support, public utility model system. PARTICIPANTS: All primary ventricular fibrillation patients seen during the study period. RESULTS: Ninety-two patients met study criteria. Data are presented as median (interquartile range). The total defibrillation interval was 9.8 minutes (7.9 to 11.8 minutes). The call-receipt to-vehicle-at-scene interval was 5.98 minutes (4.4 to 7.3 minutes). The vehicle at-scene-to-defibrillation interval was 3.6 minutes (2.5 to 4.6 minutes). CONCLUSION: The use of synchronized clocks in automated event-recording systems may provide a method of accurately measuring the time elapsed before defibrillation. PMID- 7492040 TI - Clinical safety of lidocaine in patients with cocaine-associated myocardial infarction. AB - STUDY OBJECTIVE: To evaluate the safety of lidocaine in the setting of cocaine induced myocardial infarction (MI). DESIGN: A retrospective, multicenter study. SETTING: Twenty-nine university, university-affiliated, or community hospitals during a 6-year period (total of 117 cumulative hospital-years). PARTICIPANTS: Patients with cocaine-associated MI who received lidocaine in the emergency department. RESULTS: Of 29 patients who received lidocaine in the setting of cocaine-associated MI, no patient died; exhibited bradydysrhythmias, ventricular tachycardia, or ventricular fibrillation; or experienced seizures after administration of lidocaine (95% confidence interval, 0% to 11%). CONCLUSION: Despite theoretical concerns that lidocaine may enhance cocaine toxicity, the use of lidocaine in patients with cocaine-associated MI was not associated with significant cardiovascular or central nervous system toxicity. PMID- 7492042 TI - Fellowship programs in medical toxicology. PMID- 7492041 TI - Portable hyperbaric oxygen therapy in the emergency department with the modified Gamow bag. AB - STUDY OBJECTIVE: To compare oxygen administration by means of an inflatable portable hyperbaric chamber with that through a nonrebreather mask for the elimination of carboxyhemoglobin (COHb). DESIGN: Double-crossover prospective analysis. SETTING: University emergency department, Level I trauma center. PARTICIPANTS: Twelve healthy paid adult volunteers, all smokers. INTERVENTIONS: Each subject smoked five cigarettes within 60 minutes. COHb levels were measured before and after smoking by means of cooximetry. Subjects then breathed hyperbaric and normobaric oxygen in separate trials for 40 minutes. Normobaric oxygen was administered through a nonrebreather face mask at 15 L/minute outside the Gamow bag. Hyperbaric oxygen was delivered inside the Gamow bag with a demand valve regulator mask at a pressure of 1.58 atmospheres absolute pressure (8.5 psi). Venous blood (.5 mL) was sampled every 5 minutes. The specimens were iced and assayed for COHb in triplicate. RESULTS: A significant increase in the elimination of COHb was observed for each subject in the Gamow bag (P < .05, repeated-measures ANOVA). The average half-life for COHb elimination was 27.5 +/- 1.08 minutes (mean +/- SE) (n = 10). IV access failure occurred in two patients, with incomplete data as a result. CONCLUSION: The modified Gamow bag eliminated COHb more quickly than did nonrebreather mask oxygen and proved simple to operate and maintain. No complications were noted for any of the subjects. One subject experienced claustrophobia, but it abated after the bag was inflated. PMID- 7492043 TI - American College of Emergency Physicians: Research fellowship program evaluation. PMID- 7492045 TI - Medical devices made into weapons by prisoners: an unrecognized risk. AB - STUDY OBJECTIVE: The alteration of a knee immobilizer into a sharp weapon by a prisoner prompted us to survey neighboring penal institutions to determine the frequency of such events. DESIGN: We mailed a nine-item survey to all detention facilities in Tennessee, Arkansas, and Mississippi. A second survey was sent to nonresponding institutions 6 weeks after the initial mailing. SETTING: The Regional Medical Center at Memphis, the designated facility for evaluation and treatment of prisoners from the county jail and state penitentiary. PARTICIPANTS: Survey respondents included 25 state penitentiaries, 31 county jails, 1 state minimum-security facility, 1 state maximum-security facility, 1 work-release center, 1 county detention center for drunken-driving offenders, and 1 federal penitentiary. RESULTS: Of the 81 institutions surveyed, 77% responded to one of the two mailings. Forty percent responded in the affirmative when asked whether stolen or unauthorized medical equipment from outside their institutions had been discovered among inmates. When respondents were questioned as to whether medical equipment, prescribed or not, had been used or altered in a criminal manner, 34% responded "yes." Medications and medical appliances were listed in the responses. CONCLUSION: A survey of 81 local and neighboring penal institutions in a three state area revealed that the illicit use of medicine and medical devices by prisoners is a legitimate safety concern of prison personnel and health care workers when medical care for inmates must be sought outside the security of their institutions. The modification of medical equipment into weapons by incarcerated patients, although clearly recognized as a security and safety problem by police authorities, appears to be unappreciated by health care workers providing episodic care to inmates. PMID- 7492044 TI - Use of Chinese therapies among Chinese patients seeking emergency department care. AB - We observed that many Chinese-Americans presenting to our emergency department use traditional Chinese therapies. We therefore conducted a survey using a convenience sample of first- and second-generation adult Chinese immigrants to characterize their use of traditional therapies. Seventy-six patients completed the survey; 43% had used Chinese therapy within 1 week of the ED visit. Use of Chinese therapies was common among first- and second-generation immigrants (44% and 42%, respectively). Patients claiming affiliation with a practitioner of Chinese medicine used Chinese therapies most frequently (60%). However, those lacking a traditional therapist and those associated with a Western physician also used traditional therapies frequently (38% and 41%, respectively). Many Chinese patients visit both a Western and a Chinese practitioner (17%). Emergency physicians treating Chinese-Americans should consider the possibility of concomitant use of traditional therapies. Physicians should be aware of the medical and social implications of alternative medical therapies. PMID- 7492046 TI - Emergency medicine in the Philippines. PMID- 7492047 TI - Seizures caused by ingestion of composition C-4. AB - Composition C-4 (C-4) is a plastic explosive widely used in both military and civilian settings. Ingestion of the active ingredient, RDX (cyclonite), can cause generalized seizures. Accidental and intentional C-4 (RDX) intoxications have occurred during manufacture or in wartime. In the literature the intentional ingestion of C-4 has been reported but not verified. We present a case of intentional C-4 abuse. PMID- 7492051 TI - Corneal abrasion during eyelid retraction. PMID- 7492050 TI - Anaphylaxis and leather protectors. PMID- 7492049 TI - Subgaleal hematoma with delayed proptosis and corneal ulceration. AB - We report the delayed occurrence in a child of proptosis caused by a subgaleal hematoma. A corneal ulcer resulted from the proptosis, which developed 9 days after mild head trauma. Evaluation revealed a heterozygous factor VII deficiency. The coagulopathy and, more likely, the use of the nonsteroidal antiinflammatory drug ibuprofen may have contributed to the extension of the hematoma into the orbital space. Careful follow-up of patients with subgaleal hematomas is necessary because the development of proptosis may be delayed. PMID- 7492048 TI - Dystonic reaction following recommended use of a cold syrup. AB - A healthy 3-year-old boy presented to our emergency department with sudden onset of abnormal movement. This patient had been treated for 2 days with conventional doses of an antihistamine/decongestant cold preparation for upper respiratory infection symptoms. The child was confused and restless, with posturing of limbs. Cranial nerve function was intact. Gastric lavage yielded an aspirate the same color as the cold syrup the child had ingested. A diagnosis of dystonic reaction was made, and the child was treated with benztropine, which effected resolution of his symptoms. This is the first reported case in the literature of dystonia induced by an antihistamine and treated with benztropine. PMID- 7492052 TI - Admitting mistakes. PMID- 7492053 TI - Removing constricting bands on the penis. PMID- 7492054 TI - Can we learn from history and unite as emergency physicians? PMID- 7492056 TI - Cruise ship medicine. PMID- 7492055 TI - Agitated delirium versus positional asphyxia. PMID- 7492058 TI - Guidelines on the role of nurse practitioners in emergency departments. American College of Emergency Physicians. PMID- 7492057 TI - Is "alcohol-free" beer a placebo? PMID- 7492060 TI - Agreements restricting the practice of emergency medicine. American College of Emergency Physicians. PMID- 7492059 TI - Guidelines on the role of physician assistants in emergency departments. American College of Emergency Physicians. PMID- 7492061 TI - Resurfacing of a large laryngeal wound with mucosa grafting: a combined technique using endoscopic suture and laser soldering. AB - Resurfacing of a large laryngeal wound has traditionally been accomplished via an open approach--transplanting free mucosa either onto the wound directly or upon a stent. This study proposes a new approach for wound resurfacing that combines endoscopic suturing with a laser welding technique. By means of a carbon dioxide (CO2) laser, a large (20 x 25 mm) defect in laryngeal mucosa involving the glottic and subglottic area was created in five dogs. Two of the animals served as controls and received no further treatment. In the remaining three animals the wound was resurfaced with buccal mucosal grafts fixed in place by standard suture placement and a technique of CO2 laser-assisted tissue soldering. The accessible edges of the graft were soldered endoscopically with human albumin-based tissue adhesive "activated" by low power (0.8 to 1.0 W) of a CO2 laser. All animals were examined endoscopically 1 week after surgery and sacrificed painlessly 4 weeks after surgery. The treated animals showed a viable mucosal graft with improved wound healing and voice function compared to that of the control animals. This new approach combines suture fixation of the inferior aspect of the mucosal graft, an area not well exposed endoscopically, and endoscopic laser soldering techniques for the repair of large laryngeal wounds. PMID- 7492062 TI - Innominate artery compression of the trachea: diagnosis and treatment by anterior suspension. A 25-year experience. AB - Suspension of the innominate artery to the sternum has been a widely accepted therapy for the relief of tracheal compression. Recently, reimplantation of the innominate artery has been advocated as a superior operative procedure. While generally successful, arterial transfer carries the risk of early bleeding and stroke, and the potential for late stenosis at the anastomotic site. Between 1969 and 1994, 25 infants and children at our institution received diagnoses of innominate artery compression and were treated by anterior suspension. All presented with stridor and one third had a history of suspected or proven apnea. Twenty-four children had excellent results, while 1 required resuspension after stridor returned. There were no major complications. Our series strongly supports the belief that anterior suspension of the innominate artery is a successful and reliable operation with minimal morbidity and mortality. More complex procedures are rarely indicated. PMID- 7492063 TI - Abnormalities in long latency responses to superior laryngeal nerve stimulation in adductor spasmodic dysphonia. AB - Sensorimotor responses to repeated electrical stimulation of the superior laryngeal nerve were compared in 8 patients with adductor spasmodic dysphonia (ADSD) and 11 normal controls to determine if adductor response disinhibition occurred in ADSD. Pairs of electrical pulses were presented at interstimulus intervals varying from 100 to 5,000 milliseconds (ms). Three responses were measured in thyroarytenoid muscles: ipsilateral R1 responses at 17 ms and ipsilateral and contralateral R2 responses between 60 and 75 ms. Conditioned response characteristics, the percent occurrence and percentage amplitude of initial responses, were measures of response inhibition. As a group, the patients had reduced response inhibition: their conditioned ipsilateral R1 response amplitudes were increased, as was the frequency of their conditioned contralateral muscle responses (p < or = .002) compared to normal. However, the patients' initial responses were normal in latency and frequency characteristics, demonstrating that the brain stem mechanisms for these responses were intact. These results suggest a central disinhibition of laryngeal responses to sensory input in ADSD. PMID- 7492066 TI - Second primary malignancies in the head and neck cancer patient. AB - With improvements in locoregional control, the problem of second primary malignancies is rapidly being pushed to the forefront of head and neck oncology. While the constant long-term risk for the head and neck cancer patient has been well established, efforts at prevention, early detection, and treatment have been somewhat futile. Recent advances in genetic predisposition, biomarkers, photodiagnostic imaging, and differentiation therapy may soon offer improved outcomes. The literature is reviewed and current research is presented regarding the history, epidemiology, etiology, diagnosis, and treatment of second primary malignancies. PMID- 7492064 TI - Salvage laryngectomy for unsuccessful larynx preservation therapy. AB - From 1983 to 1991, 31 patients underwent salvage laryngectomy for persistent or recurrent squamous carcinoma of the larynx (14), hypopharynx (15), or oropharynx (2) as part of a larynx preservation protocol. Laryngectomy was performed as a consequence of poor response to induction chemotherapy in 13 and for recurrent disease after completion of chemotherapy and irradiation in 18. Postoperative pharyngocutaneous fistula occurred in 39%, resulting in prolonged hospitalization. Local control was achieved in 68%, more often in patients with laryngeal as opposed to nonlaryngeal primaries (86% versus 53%; p = .05). The overall actuarial survival and disease-specific survival at 2 years were 32% and 38%, respectively. Disease-specific survival at 2 years was better in patients with laryngeal as compared to nonlaryngeal primaries (56% versus 24%; p = .02). There were no long-term survivors among the nonlaryngeal primary patients. In selected patients in whom larynx preservation failed, salvage laryngectomy was associated with acceptable local control and survival. Palliation was obtained in patients who were not cured by their laryngectomy. Future investigation will focus on identification of factors predicting complications and strategies to reduce the incidence and severity. PMID- 7492067 TI - Pneumomediastinum after rigid bronchoscopy. AB - The accumulation of extrapulmonary air is a well-known complication of airway endoscopic procedures. However, pulmonic disease alone can predispose toward pneumomediastinum and pneumothorax, without iatrogenic manipulation. In this case, a portable chest radiograph diagnosed the cause of the sudden accumulation of extrapulmonary air after rigid bronchoscopy as alveolar rupture, rather than iatrogenic airway perforation. The pathophysiology of pneumothorax and pneumomediastinum and the interpretation of chest radiographs in these situations is reviewed. PMID- 7492065 TI - Reappraisal of the monothermal warm caloric screening test. AB - The usefulness of a monothermal warm caloric screening test (MWST) for predicting alternate binaural bithermal test abnormalities has been reexamined. Previously, researchers have reported false-negative rates for the MWST ranging from 0% to over 35%. Alternate binaural bithermal test data from 504 consecutive patients were evaluated with MWST failure criteria of 24.5% and 29%, with and without the addition of failure criteria from subtests of the electronystagmography examination. Sensitivity of the MWST alone was high, and improved with additional failure criteria. The specificity levels were correspondingly high. Results support the use of the MWST with adjunctive failure criteria when patients have otherwise normal electronystagmography findings. PMID- 7492068 TI - Pendred's syndrome: a study of patients and relatives. AB - Four families, 29 members, with Pendred's syndrome were studied to clarify hearing loss and hormonal status. The ages ranged from 3 to 50 years. Complete Pendred's syndrome was found in 9 patients. They had bilateral profound hearing loss with residual hearing at low frequencies. Goiter was diagnosed at the age of 1 to 14 years with a positive perchlorate discharge test. Twelve of the patients' relatives showed partial Pendred's syndrome. Mild sensorineural hearing losses occurred in the low- and medium-range frequencies with normal perchlorate discharge test results in 6 cases. The other 6 had a slight drop in the perchlorate discharge test results with normal hearing. Five subjects were normal and 3 had normal hormonal and normal perchlorate discharge test results, but were not tested audiologically. This paper shows that patients with Pendred's syndrome may have goiter at birth or develop it between 8 and 14 years, that their deafness is bilateral and profound, and that their perchlorate discharge tests are positive. Relatives of Pendred's syndrome patients showed mild low-frequency sensorineural hearing loss without goiter and normal perchlorate discharge test results in half the cases, and a slight drop in the perchlorate discharge test results with normal hearing and without goiter in the other half. A correlation between these findings and genetic studies needs further investigation. PMID- 7492069 TI - Subglottic ductal cysts in the preterm infant: association with laryngeal intubation trauma. AB - Acquired subglottic ductal cysts following prolonged intubation in preterm infants have been reported with increasing frequency during the past two decades. This paper reviews the subglottic ductal cysts diagnosed in 15 pediatric patients between 1989 and 1993--the largest such review. Eighty percent were observed in preterm low-birth weight infants following prolonged intubation, and all but 3 of the patients had major intubation trauma of the larynx--an association not previously reported. The cysts were usually multiple and arose in the posterolateral subglottis. Most were small and did not cause significant airway obstruction, but associated intubation changes were sufficiently severe to necessitate tracheotomy in 10 patients. The cysts observed in our study did not enlarge and in 4 patients were observed to spontaneously resolve or shrink in size--trends not previously reported. Subglottic ductal cysts noted in preterm infants after prolonged intubation most commonly occur in association with laryngeal intubation trauma and are likely a component of intubation trauma. Although these cysts may resolve without therapy, careful follow-up and treatment of potentially obstructing subglottic ductal cysts is recommended. PMID- 7492070 TI - Ability of histamine to increase nasal mucosal permeability to macromolecules in guinea pigs. AB - The effect of histamine on nasal mucosal permeability against an antigen was investigated by using modified passive cutaneous anaphylaxis (PCA) reactions in normal and actively sensitized guinea pigs. The administration of a dinitrophenyl coupled Ascaris (DNP-Ascaris) solution as an antigen into the nasal cavity caused PCA reactions in the dorsal skin of normal guinea pigs. The administration of histamine into the nasal cavity before the antigen treatment significantly enhanced the anaphylactic responses. The PCA reactions did not occur when ovalbumin (OA) was administered intranasally in normal guinea pigs. In guinea pigs sensitized against DNP-Ascaris, however, PCA reactions to anti-OA antiserum were elicited by the intranasal administration of OA. The intranasal administration of histamine before the antigen treatment also enhanced anaphylactic responses in sensitized guinea pigs. These results indicate that histamine increases nasal mucosal permeability and that this may be one of the causes of nasal hypersensitivity in nasal allergy. PMID- 7492071 TI - Pneumolabyrinth as a computed tomographic finding in poststapedectomy vertigo. PMID- 7492072 TI - Oral squamous cell carcinoma and the mandible. AB - Of the various oral carcinoma subsites, the carcinomas related to the mandibular region (alveolar ridge, floor of mouth, lower retromolar area, lower buccal sulcus) are among those with the highest rate of recurrences. Neoplastic invasion of the mandible is one of the factors promoting the recurrences. Determining the presence and extent of bony involvement is therefore critical in the management of patients. The surgeon has several methods to aid in the determination of margins of invasion: radiology, gross clinical inspection, and intraoperative frozen section. Each has its deficiences and in the end, it is a combination of the methods, tempered by clinical judgment, that produces the best results. PMID- 7492073 TI - Hereditary Hearing Impairment Resource Registry. PMID- 7492074 TI - Antifungal agents in neonatal systemic candidiasis. PMID- 7492075 TI - Monoclonal antibodies to Cryptococcus neoformans glucuronoxylomannan enhance fluconazole efficacy. AB - Monoclonal antibody (MAb) 2H1, which binds to the capsular glucuronoxylomannan (GXM) of the fungus Cryptococcus neoformans, prolonged survival and decreased fungal burden in an experimental murine infection. Fluconazole (FLU) is a triazole antibiotic which is effective against C. neoformans. The efficacy of MAb 2H1 in combination with FLU was studied in vitro with the murine macrophage-like cell line J7741.16 and in vivo in mice infected intravenously. In vitro, the combination of MAb 2H1 and FLU was more effective than either agent alone in reducing the number of CFU of C. neoformans cocultured with J774.16 cells. In combination with FLU, GXM-binding MAbs of the immunoglobulin M (IgM), IgG1, IgG2a, IgG2b, IgG3, and IgA isotypes were effective in reducing the numbers of CFU in C. neoformans-J774.16 cocultures. For the in vivo experiments, A/JCr mice were infected intravenously with 5 x 10(5) organisms treated with MAb and FLU. The therapeutic effect of MAb 2H1 was primarily to reduce the number of CFU in the lung and the serum GXM level, whereas FLU was most effective in reducing the number of CFU in the brain. Mice receiving combination therapy had lower numbers of CFU in the lung and serum GXM levels than mice treated with FLU alone. Administration of MAb 2H1 with or without FLU had little or no effect on the number of CFU in the brain. The results provide support for combined therapy. PMID- 7492076 TI - SDZ PRI 053, an orally bioavailable human immunodeficiency virus type 1 proteinase inhibitor containing the 2-aminobenzylstatine moiety. AB - A series of inhibitors of human immunodeficiency virus type 1 (HIV-1) proteinase containing the 2-aralkyl-amino-substituted statine moiety as a novel transition state analog was synthesized, with the aim to obtain compounds which combine anti HIV potency with oral bioavailability. The reduced-size 2-aminobenzylstatine derivative SDZ PRI 053, which contains 2-(S)-amino-3-(R)-hydroxyindane in place of an amino acid amide, is a potent and orally bioavailable inhibitor of HIV-1 replication. The antiviral activity of SDZ PRI 053 was demonstrated in various cell lines, in primary lymphocytes, and in primary monocytes, against laboratory strains as well as clinical HIV-1 isolates (50% effective dose = 0.028 to 0.15 microM). Cell proliferation was impaired only at 100- to 300-fold-higher concentrations. The mechanism of antiviral action of the proteinase inhibitor SDZ PRI 0.53 was demonstrated to be inhibition of gag precursor protein processing. The finding that the inhibitory potency of SDZ PRI 053 in chronic virus infection, determined by p24 release, was considerably lower than that in de novo infection may be explained by the fact that the virus particles produced in the presence of SDZ PRI 053 are about 50-fold less infectious than those from untreated cultures. Upon intravenous administration, half-lives in blood of 100 and 32 min in mice and rats, respectively, were measured. Oral bioavailability of SDZ PRI 053 in rodents was 20 to 60%, depending on the dose. In mice, rats, and dogs, the inhibitor levels after oral administration remained far above the concentrations needed to efficiently block HIV replication in vitro for a prolonged period. This compound is thus a promising candidate for clinical use in HIV disease. PMID- 7492077 TI - Incidence of various gyrA mutants in 451 Staphylococcus aureus strains isolated in Japan and their susceptibilities to 10 fluoroquinolones. AB - Point mutations in the gyrA genes of 451 clinical strains of Staphylococcus aureus isolated in Japan were detected by a combination of nonradioisotopic single-strand conformation polymorphism analysis and restriction fragment length polymorphism analysis and by direct sequencing. Six types of gyrA mutations were observed in 149 of the 451 strains (33%), and ofloxacin MICs were greater than 6.25 micrograms/ml for 147 of the 149 strains (98.7%). These mutations were localized between codons 84 and 88, and they were associated with fluoroquinolone resistance. Two types of silent mutations were also found. Among these eight types of mutations, three types are novel, i.e., the serine at position 84 (Ser 84)-->Val (TCA-->GTA), Ser-84-->Leu (TCA-->TTA) plus Ile-86 (ATT-->ATC, silent), and Phe-110 (TTT-->TTC, silent). Among GyrA mutants, strains with a Ser-84-->Leu alteration and strains with a Glu-88-->Lys alteration were dominant. In contrast, few strains had Ser-84-->Ala and Glu-88-->Gly alterations. All fluoroquinolones tested showed greater than a fourfold decrease in their activities in terms of their MICs that inhibited 50% of strains tested for each GyrA mutant, in comparison with their MICs that inhibited 50% of strains tested for susceptible strains. Most of the currently available fluoroquinolones, such as norfloxacin, enoxacin, ofloxacin, ciprofloxacin, tosufloxacin, lomefloxacin, sparfloxacin, and fleroxacin, were ineffective against each mutant. Mutants containing a Ser-84- >Leu or Val alteration showed high-level resistance to fluoroquinolones, and one containing a Ser-84-->Ala alteration showed relatively low-level resistance. Double mutations were associated with a higher level of resistance than single mutations. PMID- 7492078 TI - Treatment of experimental endocarditis due to erythromycin-susceptible or resistant methicillin-resistant Staphylococcus aureus with RP 59500. AB - RP 59500 is a new injectable streptogramin composed of two synergistic components (quinupristin and dalfopristin) which are active against erythromycin-susceptible and -resistant gram-positive pathogens. The present experiments compared the therapeutic efficacy of RP 59500 with that of vancomycin against experimental endocarditis due to either of two erythromycin-susceptible or two constitutively erythromycin-resistant isolates of methicillin-resistant Staphylococcus aureus. RP 59500 had low MICs for the four test organisms as well as for 24 additional isolates (the MIC at which 90% of the isolates were inhibited was < 1 mg/liter) which were mostly inducibly (47%) or constitutively (39%) erythromycin resistant. Aortic endocarditis in rats was produced with catheter-induced vegetations. Three day therapy was initiated 12 h after infection, and the drugs were delivered via a computerized pump, which permitted the mimicking of the drug kinetics produced in human serum by twice-daily intravenous injections of 7 mg of RP 59500 per kg of body weight or 1 g of vancomycin. Both antibiotics reduced vegetation bacterial titers to below detection levels in ca. 70% of animals infected with the erythromycin-susceptible isolates (P < 0.05 compared with titers in controls). Vancomycin was also effective against the constitutively resistant strains, but RP 59500 failed against these isolates. Further experiments proved that RP 59500 failures were related to the very short life span of dalfopristin in serum (< or = 2 h, compared with > or = 6 h for quinupristin), since successful treatment was restored by artificially prolonging the dalfopristin levels for 6 h. Thus, RP 59500 is a promising alternative to vancomycin against methicillin-resistant S. aureus infections, provided that pharmacokinetic parameters are adjusted to afford prolonged levels of both of its constituents in serum. This observation is also relevant to humans, in whom the life span of dalfopristin in serum is also shorter than that of quinupristin. PMID- 7492079 TI - RP 59500 prophylaxis of experimental endocarditis due to erythromycin-susceptible and -resistant isogenic pairs of viridans group streptococci. AB - RP 59500 is a new injectable streptogramin composed of two synergistic components (quinupristin and dalfopristin) which are active against a number of erythromycin susceptible and -resistant gram-positive bacteria. The following experiments investigate the ability of RP 59500 to prevent experimental endocarditis due to either of two erythromycin-susceptible streptococcal isolates or their constitutively erythromycin-resistant Tn916 delta E transconjugants. RP 59500 had low MICs (0.125 to 0.5 mg/liter) for all four test organisms and was substantially bactericidal in vitro. Rats with catheter-induced aortic vegetations were given single-dose antibiotic prophylaxis 30 to 60 min before bacterial inoculation through a computerized pump system which permitted the simulation of drug kinetics for humans produced by either 7 mg of RP 59500 per kg of body weight or 1 g of vancomycin. Single-dose RP 59500 prophylaxis successfully prevented endocarditis due to both the erythromycin-susceptible parent strains and their erythromycin-resistant derivatives in rats challenged with the minimal inoculum infecting 90% of controls. In addition, RP 59500 also prevented infection in animals challenged with fivefold-larger inocula of the erythromycin-susceptible parent strains. Vancomycin successfully prevented endocarditis due to any of the four test organisms. These results underline the in vivo efficacy of RP 59500 against both erythromycin-susceptible and -resistant streptococci. Such good results against the resistant strains would not be expected with erythromycin or clindamycin, which are the standard macrolidelincosamide-streptogramin antibiotics used for endocarditis prophylaxis in humans. An oral form of RP 59500 which might advantageously replace some of the older prophylactic regimens is currently being developed. PMID- 7492080 TI - Mechanism of inhibition of duck hepatitis B virus polymerase by (-)-beta-L-2',3' dideoxy-3'-thiacytidine. AB - We have used the endogenous reverse transcriptase reaction of viral core particles from duck liver to elucidate the mechanism of inhibition of duck hepatitis B virus (DHBV) replication by the nucleoside analog (-)-beta-L-2',3' dideoxy-3'-thiacytidine (3TC). As is the case in human immunodeficiency virus replication, 3TC-5'-triphosphate (3TC-TP) acts as a chain terminator for the DNA polymerase activities. The results of several different experiments support this conclusion, which explains the potent activity of 3TC against the hepadnaviruses. In isolated DHBV core particles, 3TC-TP inhibited the reverse transcriptase in a manner that resembled competitive inhibition with respect to dCTP. However, the kinetics of inhibition was not linear on a double-reciprocal plot for the highest concentrations of 3TC-TP and the lowest concentration of dCTP. This anomaly would be expected if binding to the nucleotide site was followed by DNA chain termination. Calculations that used only the linear part of the curve yielded a Ki of 0.78 +/- 0.10 microM 3TC-TP. The inhibition of core particles incubated in vitro with 3TC-TP was not reversed by removal of the free inhibitor. 3TC-TP inactivated the reverse transcriptase activity in a concentration-dependent manner. The Km of the chain termination reaction was calculated at 0.71 +/- 0.05 microM. Similar competitive kinetics and irreversible inhibition were also obtained on the endogenous DNA polymerase from viral particles from serum, suggesting that 3TC-TP also acts as a chain terminator of the DNA-directed DNA polymerase of DHBV replication.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7492081 TI - Antimicrobial characterization and interrelationships of dirithromycin and epidirithromycin. AB - Dirithromycin is the 9-N,11-O-oxazine adduct formed from 9(S)-erythromycylamine and 2-(2-methoxyethoxy)acetaldehyde in which the methoxyethoxymethyl substituent on the oxazine ring possesses the R configuration. Epidirithromycin is its isomer in which the methoxyethoxymethyl substituent has the opposite (S) configuration. Both compounds readily epimerize in solution, reaching an equilibrium ratio of 85:15 in favor of dirithromycin, given sufficient time. The rate of interconversion is dependent upon pH, temperature, and solvent. An enriched sample of epidirithromycin (95% purity) was synthesized by condensing erythromycylamine and 2-(2-methoxyethoxy)acetaldehyde in diethyl ether as the reaction solvent, and the product was fully characterized by nuclear magnetic resonance spectroscopy and high-pressure liquid chromatographic (HPLC) analysis. Both oxazine derivatives readily hydrolyze to erythromycylamine, so all three compounds exhibit the same antibiotic activity in vitro. In order to determine whether dirithromycin itself possesses significant antimicrobial activity without initial hydrolysis to erythromycylmine, inhibition of cell-free ribosomal protein synthesis was measured under conditions which were adapted to minimize hydrolysis, as measured by analytical HPLC in parallel experiments. Under these particular conditions, inhibition of ribosomal protein synthesis by dirithromycin was < 10% of the value measured for erythromycylamine. PMID- 7492082 TI - Response of rat model of Pneumocystis carinii pneumonia to continuous infusion of deferoxamine. AB - The iron-chelating drug deferoxamine mesylate (DFO) is active against Pneumocystis carinii in vitro and in rat and mouse models of P. carinii pneumonia. Because DFO has a short half-life, daily divided or continuous dosage was expected to improve the dose response, as is the case with DFO treatment of malaria. Therefore, results of single daily intraperitoneal injections were compared with results of an evenly divided four-times-daily dosage and the efficacy of delivery with implanted infusion pumps. The highest bolus dosage (1,000 mg kg-1 of body weight day-1) was as effective as the standard combination of trimethoprim with sulfamethoxazole. Unexpectedly, very little improvement was observed with the divided or continuous dosage, and several mechanisms that could account for this are discussed. PMID- 7492083 TI - Disposition of S-1108, a new oral cephem antibiotic, and metabolic fate of pivalic acid liberated from [pivaloyl-14C]S-1108 in rats and dogs. AB - [pivaloyl-14C]S-1108, which is 14C labeled at the pivalic acid moiety of the pivaloyloxymethyl side chain of S-1108, was administered orally to rats and dogs, and the disposition of pivalic acid cleft from S-1108 was examined. Besides pivaloylcarnitine and pivaloylglucuronide, pivaloylglycine was identified in dog urine as a metabolite of pivalic acid by thin-layer chromatography and high performance liquid chromatography analysis. The concentrations in the plasma of rats to which doses of 6.65, 26.6, and 532 mg/kg of body weight were administered showed dose-proportionate levels. The radioactivity was eliminated rapidly, with a half-life of approximately 3 h until 24 h at both the 6.65- and 26.6-mg/kg doses. Free pivalic acid in plasma accounted for more than 80% of the concentration of radioactivity. Radioactivity was distributed throughout the body and was eliminated quickly at a rate similar to that of radioactivity from plasma. Most of the absorbed radioactivity was excreted in the urine, and it was completed within 24 h after administration. In dogs, the half-life of radioactivity in plasma was longer than that in the rats. The ratio of free pivalic acid in plasma was 60 to 70% of the radioactivity in plasma. The concentration of radioactivity in the liver, cortex of the kidney, and skeletal muscle 144 h after oral dosing was more than 10 times higher than the concentration in plasma for all doses. Urinary excretion in dogs was slower than that in rats. The differences in the disposition of pivalic acid between dogs and rats may account for differences in the degree of skeletal muscle disorders. The safety in humans of S-1108 given at 200 mg three times a day is discussed in relation to the metabolic formation of the carnitine conjugate of pivalic acid and the reduction of the carnitine concentration in plasma. PMID- 7492085 TI - Purification and characterization of a new beta-lactamase from Bacteroides uniformis. AB - A beta-lactam-resistant Bacteroides uniformis strain was isolated from a clinical specimen. The strain produced large amounts of beta-lactamase and was resistant to penicillins and cephalosporins. The specific activity of the unpurified beta lactamase was 4.8 U/mg of protein with nitrocefin as the substrate. The enzyme was purified 188-fold by Q-Sepharose, Sephacryl S-300, and Mono Q column passages. Kinetic parameters of the enzyme were determined by a micromethod performed in microtiter plates. beta-Lactamase was inhibited by cefoxitin and imipenem and hydrolyzed cephalosporins more rapidly than penicillins. The molecular weight was determined by sodium dodecyl sulfate-gradient gel electrophoresis to be 32,500, and the isoelectric point was 4.5. PMID- 7492084 TI - Susceptibilities of nontuberculosis mycobacterial species to amoxicillin clavulanic acid alone and in combination with antimycobacterial agents. AB - Neither amoxicillin nor clavulanic acid used alone was active at the highest level tested, i.e., 256.0 micrograms/ml, in vitro against 24 isolates of Mycobacterium fortuitum, Mycobacterium kansasii, and Mycobacterium marinum. However, the MIC of an amoxicillin-clavulanic acid combination of 2:1 was < or = 8.0/4.0 micrograms/ml for 50 percent of the isolates tested, with all isolates being inhibited in the range of 4.0/2.0 to 32.0/16.0 micrograms/ml, respectively. Titration of amoxicillin-clavulanic acid with a fixed 2-micrograms/ml concentration of ethambutol resulted in synergistic activity against 3 of 9 isolates of M. fortuitum, 10 of 10 isolates of M. kansasii, and 5 of 5 isolates of M. marinum. This observation was confirmed in a checkerboard analysis in which fractional inhibitory concentrations were < or = 0.5 for 20 of the 24 isolates. Synergistic activity was observed against the other four isolates in one of two trials. On the other hand, titration of amoxicillin-clavulanic acid in the presence of either one or two fixed concentrations of isoniazid, rifampin, cycloserine, tetracycline, or amikacin failed to result in synergism. PMID- 7492086 TI - Therapeutic efficacy of a polymyxin B-dextran 70 conjugate in experimental model of endotoxemia. AB - Numerous studies have suggested that lipopolysaccharide (LPS), a major component of the cell wall of gram-negative bacteria, is responsible for the initiation of gram-negative septic shock. Previously, others have designed therapeutic regimens to target the biologically active lipid A region of LPS by either neutralization of the biological properties of LPS or enhancement of clearance of this molecule. One such compound capable of neutralizing lipid A is the antibiotic polymyxin B. However, the clinical utility of polymyxin B is limited by its toxicity. We therefore covalently conjugated this antibiotic to the high-molecular-weight polysaccharide dextran 70, resulting in reduced toxicity of polymyxin B but retention of its endotoxin-neutralizing ability. The studies described in this report were designed to test the in vivo efficacy of this compound in an experimental animal model of gram-negative septic shock. Mice were administered graded doses of Escherichia coli or Pseudomonas aeruginosa along with D galactosamine and the antibiotic imipenem. We had previously determined that antibiotic chemotherapy provides significant protection against E. coli-mediated lethality with smaller doses of bacteria; however, the antibiotic does not provide protection against larger doses of bacteria, but it is effective at killing the bacterial inoculum in vivo. Administration of the polymyxin B-dextran 70 conjugate provided significant protection when given with an antibiotic but was not effective by itself. A requirement for a pretreatment period prior to E. coli challenge was shown to depend upon the bacterial challenge dose. In other studies using this D-galactosamine sensitization model, we demonstrated that the lipid A-specific conjugate had no effect on lethality caused by staphylococcus aureus or tumor necrosis factor alpha. The results of these studies indicate that this compound is effective in preventing lethal gram-negative septic shock in mice and may be useful as a potential therapeutic agent in humans as well. PMID- 7492088 TI - Antibacterial activity of WY-49605 compared with those of six other oral agents and selection of disk content for disk diffusion susceptibility testing. AB - The in vitro antimicrobial activity of an oral penem, WY-49605, was compared with those of six other oral antimicrobial agents against 598 bacterial isolates representing 51 different species. WY-49605 exhibited good activity against most gram-positive bacteria and members of the family Enterobacteriaceae. It had little activity against nonfermenting gram-negative bacilli, Enterobacter spp., Serratia spp., enterococci, Staphylococcus haemolyticus, and methicillin resistant Staphylococcus aureus. Its activity was unaffected by the beta lactamases of Neisseria gonorrhoeae, Haemophilus influenzae, and staphylococci. Disk diffusion susceptibility tests were performed with 5-, 10-, 15-, and 30 micrograms WY-49605 disks. The 5-micrograms disk is recommended, with tentative breakpoints of > or = 16 mm for susceptibility (MIC, < or = 2.0 microgram/ml) and < or = 12 mm for resistance (MIC, > or = 8.0 micrograms/ml). PMID- 7492087 TI - Contribution of the C-8 substituent of DU-6859a, a new potent fluoroquinolone, to its activity against DNA gyrase mutants of Pseudomonas aeruginosa. AB - Inhibitory effects of five quinolones against DNA gyrases purified from four quinolone-resistant clinical isolates of Pseudomonas aeruginosa and the quinolone susceptible strain PAO1 were examined. All of the quinolone-resistant strains tested were found to be DNA gyrase mutants. The 50% inhibitory concentrations (IC50s) of the quinolones for these DNA gyrases roughly correlated with their MICs. Interestingly, gyrase inhibition by DU-6859a was found to be significantly less affected by these mutations that inhibition by other currently available quinolones. To assess the enhanced activity shown by DU-6859a, the effects of quinolones with altered substituents at the N-1, C-7, and C-8 positions of the quinolone ring of DU-6859a were tested. Measurement of MICs for four DNA gyrase mutants and IC50s for their purified DNA gyrases showed that removal of the C-8 chlorine of DU-6859a significantly increased MICs and IC50s for DNA gyrase mutants. However, no deleterious effects were observed when either the fluorine on the cyclopropyl substituent at the N-1 position or the cyclopropyl ring at the C-7 substituent was removed. Moreover, removal of the C-8 chlorine also increased the MIC for 19 of 20 quinolone-resistant clinical isolates. Our results led to the conclusion that DU-6859a is much more active against quinolone-resistant clinical isolates of P. aeruginosa than other currently available quinolones, probably because of its strong inhibitory effects against mutant quinolone resistant DNA gyrases, and that the C-8 chlorine is necessary for these potent effects. PMID- 7492089 TI - Inducible and constitutive expression of vanC-1-encoded resistance to vancomycin in Enterococcus gallinarum. AB - Clinical isolates Enterococcus gallinarum AIB39 and E. gallinarum GS1 were studied to establish whether the expression of vanC-1-mediated resistance may be inducible or constitutive. By growth curve analysis, strain AIB39 exhibited the same lag period (i.e., 1 to 1.5 h) whether it was subcultured to unsupplemented brain heart infusion broth or broth containing 6 micrograms of vancomycin per ml, a growth pattern typical of constitutively expressed resistance. Use of high performance liquid chromatography (HPLC) to separate peptidoglycan precursor extracts substantiated this finding because the serine-terminating pentapeptide precursor UDP-MurNAc-L-Ala-D-Glu-L-Lys-D-Ala-D-Ser was produced in the presence and absence of vancomycin, whereas no UDP-MurNAc-L-Ala-D-Glu-L-Lys-D-Ala-D-Ala was detected. In contrast, results with strain GS1 were consistent with inducible expression. GS1 demonstrated a lag time that was 3 to 4 h longer when it was subcultured to vancomycin-containing broth than when it was subcultured in unsupplemented broth. HPLC analysis showed that in the absence of vancomycin only UDP-MurNAc-L-Ala-D-Glu-L-Lys-D-Ala-D-Ala was detected, but in the presence of drug only UDP-MurNAc-L-Ala-D-Glu-L-Lys-D-Ala-D-Ala was found. Inducible expression of vanC-1-mediated resistance in E. gallinarum is consistent with recent findings suggesting the presence of at least two ligases in this species. Although vanC-1 may be intrinsic to E. gallinarum, our findings raise doubt regarding the natural mechanism of this gene's expression. PMID- 7492090 TI - Evaluation of retinal toxicity and efficacy of the anticytomegalovirus compound 2 amino-7-[(1,3-dihydroxy-2-propoxy)methyl]purine. AB - Compound 2242, also known as 2-amino-7-[(1,3-dihydroxy-2-propoxy)methyl]purine, is the first known antivirally active nucleoside analog with the side chain substituted at the N-7 position of the purine ring system. Our purpose was to evaluate its retinal toxicity and assess the efficacy of its highest nontoxic concentration in a rabbit model of herpes simplex retinitis. Concentrations of the drug from 0.5 to 2,000 microM were injected intravitreally in twelve New Zealand White rabbits. Fundoscopic, histologic, and electrophysiologic data revealed no evidence of toxicity even at the highest dose of the compound. Dutch pigmented rabbits (n = 34) had their left eyes injected with herpes simplex virus type 1 3 days after, concurrently, or 3 days before intravitreal injection of either 2,000 microM compound 2242 or 480 microM ganciclovir (final concentration in the eye). Both compound 2242 and ganciclovir were equally effective compared with saline when administered simultaneously with the virus (P < 0.0001). In the 3-day pretreatment paradigm, compound 2242 was superior to ganciclovir (P < 0.04), but there was no clear difference between the two with regard to their effects on an established infection. The pharmacokinetics of compound 2242 in 10 rabbits injected intravitreally with 30 microM showed an intravitreal half-life of 8 h. This compound, which may be orally active in its pro form, has a very high therapeutic index in the eye and is more efficient than ganciclovir in this animal model of herpes retinitis. PMID- 7492091 TI - Mechanism of action of antimycobacterial activity of the new benzoxazinorifamycin KRM-1648. AB - The mechanism of antimicrobial activity of KRM-1648 (KRM), a new rifamycin derivative with potent antimycobacterial activity, was studied. Both KRM and rifampin (RMP) inhibited RNA polymerases from Escherichia coli and Mycobacterium avium at low concentrations: the 50% inhibitory concentrations (IC50s) of KRM and RMP for E. coli RNA polymerase were 0.13 and 0.10 micrograms/ml, respectively, while the IC50s for M. avium RNA polymerase were 0.20 and 0.07 microgram/ml. Both KRM and RMP exerted weak inhibitory activity against Mycobacterium fortuitum RNA polymerase, rabbit thymus RNA polymerases, E. coli DNA polymerase I, and two types of reverse transcriptases. Uptake of 14C-KRM by M. avium reached 18,000 dpm/mg (dry weight) 1.5 h after incubation, while uptake by E. coli cells was slight. KRM was much more effective in inhibiting uptake of 14C-uracil than was RMP (IC50 of KRM, 0.04 microgram/ml; IC50 of RMP, 0.12 microgram/ml). These findings suggest, first, that the potent antimycobacterial activity of KRM is due to inhibition of bacterial RNA polymerase and, second, that the activity of KRM against target organisms depends on target cell wall permeability. PMID- 7492092 TI - Pharmacokinetics of ofloxacin in serum and vitreous humor of albino and pigmented rabbits. AB - The purpose of this study was to evaluate the pharmacokinetics of ofloxacin in serum and vitreous humor samples from albino and pigmented rabbits by using a recently described animal model which permits robust estimation of parameter values. The drug was administered to rabbits intravenously, multiple vitreous humor and serum samples were taken from each rabbit, and the vitreous humor and serum samples were assayed by high-pressure liquid chromatography. The pharmacokinetic parameters were determined with RSTRIP, an iterative, nonlinear, weighted, least-squares regression program. Eight New Zealand White rabbits and eight Dutch Belted rabbits (split into single-dose and multiple-dose groups) were investigated in this study. The value of penetration into the vitreous humor of albino rabbits (n = 6) was 32.6% +/- 2.12%, with terminal-elimination half-life values of 3.21 and 2.39 h, respectively, for vitreous humor and serum. In pigmented rabbits after a single dose (n = 3) and with a steady-state concentration of drug in serum (n = 4), penetration values were similar, at 30.4% +/- 2.98% and 30.0% +/- 4.12%, respectively (P > 0.10). Following a single dose of ofloxacin, pigmented animals had elimination half-life values from serum and vitreous humor of 2.64 and 4.32 h, respectively. After steady state was achieved, half-life values for serum and vitreous humor were 3.12 and 6.05 h, respectively.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7492093 TI - Activities of oral and parenteral agents against penicillin-susceptible and resistant pneumococci. AB - This study examined bacteriostatic and bactericidal activities of oral and parenteral antibiotics for penicillin-susceptible and intermediately and fully penicillin-resistant pneumococci. beta-Lactamase inhibitors did not affect beta lactam results. The activities of ampicillin, amoxicillin +/- clavulanate, WY 49605, cefuroxime, cefpodoxime, cefdinir, cefixime, and cefaclor against two penicillin-susceptible, two intermediately penicillin-resistant, and two fully penicillin-resistant pneumococcal strains were tested. For all three groups, bacteriostatic values of amoxicillin and WY-49605 were lower than were those of other beta-lactams tested. Of the cephalosporins, cefdinir, cefuroxime, and cefpodoxime yielded the lowest bacteriostatic values. All beta-lactams were bactericidal (reduced original counts by > or = 3 log10 CFU/ml) at 1 dilution above bacteriostatic values, except for cefpodoxime (bactericidal at 2 dilutions above bacteriostatic values for one susceptible strain and one intermediately resistant strain), cefuroxime (bactericidal at 2 dilutions above bacteriostatic values for one intermediately resistant strain), and ampicillin (bactericidal at 2 dilutions above bacteriostatic values for one intermediately resistant strain). The activities of piperacillin, piperacillin-tazobactam, ticarcillin, ticarcillin clavulanate, ampicillin, ampicillin-sulbactam, ceftriaxone, ceftazidime, and ciprofloxacin against four penicillin-susceptible, two intermediately penicillin resistant, and four fully penicillin-resistant pneumococcal strains were evaluated. Bacteriostatic values of piperacillin, ampicillin, and ceftriaxone for all groups were lower than were those of ticarcillin and ceftazidime. Bacteriostatic values of ciprofloxacin were unaffected by penicillin susceptibility. All beta-lactams were bactericidal at 1 dilution above the bacteriostatic value, except for piperacillin (bactericidal at 2 dilutions above the bacteriostatic value for one intermediately resistant strain), ticarcillin (bactericidal at 2 dilutions above the bacteriostatic value for one susceptible strain and one resistant strain), ampicillin (bactericidal at 2 dilutions above the bacteriostatic value for two resistant strains), ceftriaxone (bactericidal at 2 dilutions above the bacteriostatic value for one resistant strain), and ceftazidime (bactericidal at 2 dilutions above the bacteriostatic value for one susceptible strain). PMID- 7492094 TI - Pharmacodynamics of RP 59500 alone and in combination with vancomycin against Staphylococcus aureus in an in vitro-infected fibrin clot model. AB - The bactericidal activity and emergence of resistance to RP 59500 (quinupristin/dalfopristin) when it was administered alone and in combination with vancomycin against fibrin clots that have been infected with methicillin susceptible Staphylococcus aureus ATCC 25923 or methicillin-resistant S. aureus (MRSA) 67 were evaluated in an in vitro pharmacodynamic infected fibrin clot model. Fibrin clots were infected with S. aureus to achieve an inoculum of approximately 10(9) CFU/g. Antibiotics were administered to simulate pharmacokinetics in humans: RP 59500 (7.5 mg/kg of body weight) every 8 h and vancomycin (15 mg/kg) every 12 h over 72 h. Preliminary test tube time-kill experiments with an inoculum of approximately 10(5) CFU/ml suggested that RP 59500 was more rapid in achieving a 99.9% reduction in the number of CFU per milliliter than vancomycin against ATCC 25923 (6.94 versus 24 h; P = 0.0003) and MRSA 67 (6.77 versus 17.03 h; P = 0.004). At a higher inoculum (approximately 10(8) CFU/ml), 99.9% kill was achieved only with the combination regimen against ATCC 25923 and MRSA 67 (10.9 and 10.5 h, respectively), with total reductions of 6.35 and 6.33 log10 CFU/ml over 24 h, respectively. In the fibrin clot model, RP 59500 was more effective than vancomycin in reducing organism titers over 72 h. In the fibrin clot model, the most optimal therapy was the combination regimen. PMID- 7492095 TI - Patterns of in vitro activity of itraconazole and imidazole antifungal agents against Candida albicans with decreased susceptibility to fluconazole from Spain. AB - Two groups of recent clinical isolates of Candida albicans consisting of 101 isolates for which fluconazole MICs were < or = 0.5 microgram/ml (n = 50) and > or = 4.0 micrograms/ml (n = 51), respectively, were compared for their susceptibilities to fluconazole, clotrimazole, miconazole, ketoconazole, and itraconazole. Susceptibility tests were performed by a photometer-read broth microdilution method with an improved RPMI 1640 medium supplemented with 18 g of glucose per liter (RPMI-2% glucose; J. L. Rodriguez-Tudela and J. V. Martinez Suarez, Antimicrob. Agents Chemother. 38:45-48, 1994). Preparation of drugs, basal medium, and inocula was done by the recommendations of the National Committee for Clinical Laboratory Standards. The MIC endpoint was calculated objectively from the turbidimetric data read at 24 h as the lowest drug concentration at which growth was just equal to or less than 20% of that in the positive control well (MIC 80%). In vitro susceptibility testing separated azole susceptible strains from the strains with decreased susceptibilities to azoles if wide ranges of concentrations (20 doubling dilutions) were used for ketoconazole, miconazole, and clotrimazole. By comparison with isolates for which fluconazole MICs were < or = 0.5 microgram/ml, those isolates for which fluconazole MICs were > or = 4.0 micrograms/ml were in general less susceptible to other azole drugs, but different patterns of decreased susceptibility were found, including uniform increases in the MICs of all azole derivatives, higher MICs of several azoles but not others, and elevated MICs of fluconazole only. On the other hand, decreased susceptibility to any other azole drug was never found among strains for which MICs of fluconazole were lower. PMID- 7492096 TI - In vitro and in vivo antifungal activities of DU-6859a, a fluoroquinolone, in combination with amphotericin B and fluconazole against pathogenic fungi. AB - DU-6859a is an investigational fluoroquinolone agent with potent bactericidal activity, but by itself it has no antifungal activity. When combined with amphotericin B (AmB), however, DU-6859a clearly enhanced the in vitro antifungal activity of AmB against Candida albicans, Candida tropicalis, Candida krusei, Candida glabrata, and Cryptococcus neoformans in microdilution checkerboard studies. Positive interactions of DU-6859a with AmB against Aspergillus fumigatus were dependent on the medium used; yeast nitrogen base supplemented with amino acids, ammonium sulfate, and 1% glucose was better for demonstrating synergism, while in RPMI 1640 medium, unexpected antagonism between the drugs occurred against three of the strains tested. In combination with fluconazole (Flu), DU 6859a increased the activity of Flu against C. albicans both in synthetic amino acid medium fungal and in supplemented yeast nitrogen base. An in vitro time-kill study revealed that DU-6859a combined with AmB significantly suppressed the regrowth of C. albicans compared with the suppression brought about by AmB used alone in a concentration-dependent fashion. Furthermore, in a model of C. albicans infection in mice, the fungal load in infected kidneys was significantly less in mice given the combination treatment of DU-6859a plus either AmB or Flu, and thus, the combination treatment resulted in prolonged survival of infected mice compared with treatment with either antifungal alone. The prolonged survival in mice given the combined treatment was also observed in mice with A. fumigatus infection, indicating that DU-6859a potentiated the actions of the antifungal agents in vivo as well as in vitro. PMID- 7492097 TI - Improved bactericidal activity of Q-35 against quinolone-resistant staphylococci. AB - The bactericidal effects of Q-35, sparfloxacin, tosufloxacin, and ofloxacin on 18 strains of methicillin-resistant Staphylococcus aureus (MRSA) and 3 strains of Staphylococcus epidermidis were studied by a viable-count method. Staphylococci as used in this study were clearly divided into two groups with respect to their susceptibilities to sparfloxacin. MICs of Q-35 and tosufloxacin were 0.05 to 0.78 microgram/ml for sparfloxacin-susceptible strains (MICs, 0.05 to 0.2 microgram/ml) and 1.56 to 12.5 micrograms/ml for sparfloxacin-resistant strains (6.25 to 25 micrograms/ml). All the sparfloxacin-resistant strains of MRSA tested contained the gyrA mutation at codon 84. Time-kill studies showed that Q-35 decreased the viable counts from approximately 10(7) CFU/ml to 10(3) to 10(5) CFU/ml within 3 h at concentrations greater than the MICs against both sparfloxacin-susceptible and -resistant strains. In contrast, sparfloxacin, tosufloxacin, and ofloxacin produced bacteriostatic effects at 3 h after exposure against sparfloxacin-resistant strains at concentrations which were greater than the respective MICs, whereas these quinolones were bactericidal against sparfloxacin-susceptible strains. The rapid bactericidal activities of Q-35 against sparfloxacin-resistant MRSA were reduced when the methoxy group of Q-35 at the 8 position was substituted with fluorine or hydrogen. Thus, our data suggest that the introduction of a methoxy group into the 8 position of quinolones contributes to the bactericidal activities of fluoroquinolones against quinolone-resistant staphylococci. PMID- 7492098 TI - Cross-resistance to polyene and azole drugs in Cryptococcus neoformans. AB - Fluconazole was observed to inhibit sterol 14 alpha-demethylase in the human pathogen Cryptococcus neoformans, and accumulation of a ketosteroid product was associated with growth arrest. A novel mechanism(s) of azole and amphotericin B cross-resistance was identified, unrelated to changes in sterol biosynthesis, as previously identified in Saccharomyces cerevisiae. Reduced cellular content of drug could account for the resistance phenotype, indicating the possible involvement of a mechanism similar to multidrug resistance observed in higher eukaryotes. PMID- 7492099 TI - In vitro assays elucidate peculiar kinetics of clindamycin action against Toxoplasma gondii. AB - In order to characterize the delayed effect of clindamycin and macrolide antibiotics against Toxoplasma gondii tachyzoites (E. R. Pfefferkorn and S. E. Borotz, Antimicrob. Agents Chemother. 38:31-37, 1994), we have carefully examined the replication of parasites as a function of time after drug addition. Intracellular tachyzoites treated with up to 20 microM clindamycin (> 1,000 times the 50% inhibitory concentration) exhibit doubling times indistinguishable from those of controls (approximately 7 h). Drug-treated parasites emerge from infected cells and establish parasitophorous vacuoles inside new host cells as efficiently as untreated controls, but replication within the second vacuole is dramatically slowed. Growth inhibition in the second vacuole does not require continued presence of drug, but it is dependent solely on the concentration and duration of drug treatment in the first (previous) vacuole. The susceptibility of intracellular parasites to nanomolar concentrations of clindamycin contrasts with that of extracellular tachyzoites, which are completely resistant to treatment, even through several cycles of subsequent intracellular replication. This peculiar phenotype, in which drug effects are observed only in the second infectious cycle, also characterizes azithromycin and chloramphenicol treatment, but not treatment with cycloheximide, tetracycline, or anisomycin. These findings provide new insights into the mode of clindamycin and macrolide action against T. gondii, although the relevant target for their action remains unknown. PMID- 7492100 TI - Inhibition of sterol 4-demethylation in Candida albicans by 6-amino-2-n pentylthiobenzothiazole, a novel mechanism of action for an antifungal agent. AB - The effects of 6-amino-2-n-pentylthiobenzothiazole (APB), a new antifungal agent, on ergosterol biosynthesis in Candida albicans and Saccharomyces cerevisiae were studied, using [14C]acetate incorporation. In C. albicans, the inhibition of growth was accompanied by a marked inhibition of acetate incorporation in 4 desmethylsterols, with a significant portion of the radiolabel being incorporated in 4,4-dimethylsterols, lanosterol, and 4,4-dimethylzymosterol and minor amounts being incorporated in 4-methylsterols and squalene. The data are interpreted as evidence of a block of the ergosterol biosynthesis pathway at the level of 4 demethylation of 4,4-dimethylzymosterol, with partial inhibition of lanosterol 14 dimethylation and squalene epoxidation also being possible. In 6-amino-2-n pentylthiobenzothiazole-treated S. cerevisiae, a significant amount of the radiolabel was incorporated also in 4-methylsterols, 4-methylzymosterol, and 4 methylfecosterol, indicating that in this microorganism there are different sensitivities of the two 4-demethylations and that the pathway is blocked at the level of 4-demethylation of 4-methylsterols. PMID- 7492101 TI - Synergistic activities of clarithromycin and antituberculous drugs against multidrug-resistant Mycobacterium tuberculosis. AB - The rise of multidrug-resistant Mycobacterium tuberculosis has complicated therapy for tuberculosis and led us to search for a potentially active combination of drugs against these strains. The susceptibilities of 12 strains of multidrug-resistant M. tuberculosis to standard antituberculous drugs (isoniazid, rifampin, ethambutol, and pyrazinamide), clarithromycin, and its metabolite, 14 hydroxyclarithromycin, were determined by use of the BACTEC radiometric method. All strains were resistant to at least two of the antituberculous drugs. Clarithromycin and 14-hydroxyclarithromycin MICs were in the range indicating resistance at > or = 8.0 micrograms/ml for all strains. Combination testing by the BACTEC method was performed with various concentrations of isoniazid, rifampin, and ethambutol, and with clarithromycin/14-hydroxyclarithromycin at fixed concentrations of 2.0/0.5 micrograms/ml, respectively. Addition of clarithromycin/14-hydroxyclarithromycin to these antituberculous drug mixtures resulted in a 4- to 32-fold reduction in MICs of isoniazid, rifampin, and ethambutol and made resistant strains susceptible. Fractional inhibitory concentrations ranged from 0.23 to 0.50 for all strains, suggesting a synergistic interaction between standard antituberculous drugs and clarithromycin/14 hydroxyclarithromycin. The ability of clarithromycin/14-hydroxyclarithromycin to enhance the activities of isoniazid, ethambutol, and rifampin in vitro suggests that this combination may be efficacious in the treatment of multidrug-resistant M. tuberculosis infections. PMID- 7492102 TI - Valaciclovir compared with acyclovir for improved therapy for herpes zoster in immunocompetent adults. AB - Acyclovir treatment of acute herpes zoster speeds rash healing and decreases pain and ocular complications. The limited oral bioavailability of acyclovir necessitates frequent dosing. Valaciclovir, the l-valyl ester of acyclovir, is rapidly and almost completely converted to acyclovir in vivo and gives three- to fivefold increases in acyclovir bioavailability. In a randomized, double-blind, multicenter study, the safety and efficacy of oral valaciclovir given at a dosage of 1,000 mg three times daily for 7 or 14 days and oral acyclovir given at a dosage of 800 mg five times daily for 7 days were compared in immunocompetent adults aged > or = 50 years with herpes zoster. Patients were evaluated for 6 months. The intent-to-treat analysis (1,141 patients) showed that valaciclovir for 7 or 14 days significantly accelerated the resolution of herpes zoster associated pain (P = 0.001 and P = 0.03, respectively) compared with acyclovir; median pain durations were 38 and 44 days, respectively, versus 51 days for acyclovir. Treatment with valaciclovir also significantly reduced the duration of postherpetic neuralgia and decreased the proportion of patients with pain persisting for 6 months (19.3 versus 25.7%). However, there were no differences between treatments in pain intensity or quality-of-life measures. Cutaneous manifestations resolved at similar rates in all groups. Adverse events were similar in nature and prevalence among groups, and no clinically important changes occurred in hematology or clinical chemistry parameters. Thus, in the management of immunocompetent patients > or = 50 years of age with localized herpes zoster, valaciclovir given at 1,000 mg three times daily for 7 days accelerates the resolution of pain and offers simpler dosing, while it maintains the favorable safety profile of acyclovir. PMID- 7492103 TI - Analysis of gyrA and grlA mutations in stepwise-selected ciprofloxacin-resistant mutants of Staphylococcus aureus. AB - Fluoroquinolone-resistant mutants were obtained in vitro from Staphylococcus aureus RN4220 by stepwise selection on increasing concentrations of ciprofloxacin. Results from sequence analysis of the quinolone resistance determining region of GyrA and of the corresponding region of GrlA, the DNA topoisomerase IV subunit, showed an alteration of Ser-80 to Tyr (corresponding to Ser-83 of Escherichia coli GyrA) or Glu-84 to Lys in GrlA of both low- and high level quinolone-resistant mutants. Second-step mutants were found to have, in addition to a mutation in grlA, reduced accumulation of norfloxacin or an alteration in GyrA at Ser-84 to Leu or Glu-88 to Lys. Third-step mutants derived from second-step mutants with reduced accumulation were found to have a mutation in gyrA. The results from this study demonstrated that mutations in gyrA or mutations leading to reduced drug accumulation occur after alteration of GrlA, supporting the previous findings (L. Ferrero, B. Cameron, B. Manse, D. Lagneaux, J. Crouzet, A. Famechon, and F. Blanche, Mol. Microbiol. 13:641-653, 1994) that DNA topoisomerase IV is a primary target of fluoroquinolones in S. aureus. PMID- 7492104 TI - Safety, pharmacokinetics, and antiviral activity of A77003, a C2 symmetry-based human immunodeficiency virus protease inhibitor. AB - A77003, an inhibitor of the human immunodeficiency virus type 1 (HIV-1) protease, was administered to asymptomatic HIV-1-infected patients in a phase I trial. The drug was given by continuous intravenous infusion at dosages of 0.035, 0.07, 0.14, and 0.28 mg/kg of body weight per h. The drug was given first for 24 h and then for up to an additional 4 weeks in a second infusion period following at least a 6-day washout. Apart from reversible increases in hepatic transaminase levels in some patients, no systemic toxicities occurred during extended infusion of the drug. Dose-related local vein irritation, despite dilution of the infusate, however, caused severe infusion site phlebitis precluding dosage escalation beyond 0.28 mg/kg/h. Pharmacokinetic analysis demonstrated dose linear increases in mean steady-state concentrations. However, clearance of the drug from plasma was unexpectedly high, averaging 62 liters/h across all groups. The concentrations of A77003 in plasma achieved the in vitro 50% inhibitory concentration (0.16 microgram/ml) only in the 0.28-mg/kg/h dosage group, but it did not attain the 90% inhibitory concentration (0.48 micrograms/ml). No statistically significant effect on CD4 cell numbers occurred in any of the groups, and there was no evidence of antiviral activity, as determined by HIV-1 p24 antigen level, quantitative plasma and cell culture, and quantitation of viral RNA in plasma. In conclusion, A77003, as formulated in the present study, causes severe phlebitis, which prevents administration of the infusates necessary to achieve high concentrations of the drug in plasma. In conclusion, A77003, as formulated in the present study, causes severe phlebitis, which prevents administration of the infusates necessary to achieve high concentrations of the drug in plasma. The lack of antiviral activity observed in the study may be a consequence of the low concentrations in plasma in all groups. PMID- 7492105 TI - In vitro and in vivo effects of penicillin and clindamycin on expression of group A beta-hemolytic streptococcal capsule. AB - Encapsulation of group A beta-hemolytic streptococci (GABHS) is an important virulence factor. The changes that occur in the frequency of encapsulation of GABHS during pharyngotonsillitis, in 20 patients treated with penicillin and 20 treated with clindamycin, were investigated. The effects of subinhibitory concentrations of these agents were also evaluated in vitro. At day 4, 8 of 10 (80%) GABHS isolates recovered from children treated with penicillin were encapsulated, compared with 1 of 5 (20%) of those from children treated with clindamycin (P < 0.05). Two days following 10 days of therapy, GABHS was eliminated from 13 of the 20 (65%) children treated with penicillin and from all treated with clindamycin (P < 0.05). At that time, six of the seven GABHS isolates recovered in patients treated with penicillin were encapsulated. GABHS were not detected after 4 days of therapy in those treated with clindamycin. Incubation of GABHS isolates with one-half of the MIC of clindamycin reduced the frequency of encapsulation, compared with that after incubation with one-half of the MIC of penicillin (12.5 versus 67.5%). These data illustrate the superiority of clindamycin over penicillin in reducing the expression of a capsule by GABHS. PMID- 7492106 TI - Product of fosC, a gene from Pseudomonas syringae, mediates fosfomycin resistance by using ATP as cosubstrate. AB - Pseudomonas syringe PB-5123, a producer of fosfomycin, is resistant to high concentrations of the antibiotic. Two possible mechanisms of resistance have been detected: (i) impermeability to exogenous fosfomycin, even in the presence of sugar phosphate uptake inducers, and (ii) antibiotic phosphorylation. The gene responsible for this last activity, fosC, encodes a ca. 19,000-Da protein and is immediately followed by a second open reading frame, which shows sequence similarities to glutathione S-transferases. FosC uses ATP as a cosubstrate in an inactivation reaction that can be reversed with alkaline phosphatase. Other nucleotide triphosphates cannot be substituted for ATP in this reaction. No relationship between fosC and the previously described genes of fosfomycin resistance was found. PMID- 7492107 TI - Influence of pentoxifylline and its derivatives on antibiotic uptake and superoxide generation by human phagocytic cells. AB - Pentoxifylline modulates multiple activities of stimulated polymorphonuclear neutrophils (PMNs), including the respiratory burst response and membrane transport of certain substances (e.g., nucleosides). We found that several weakly basic antibiotics are highly concentrated by human PMNs and that these drugs also inhibit the respiratory burst response (by a mechanism different from that of pentoxifylline). Since both pentoxifylline and antibiotics will be administered to some patients with serious infections, we have evaluated several types of interactions between these drugs and human PMNs and have attempted to identify the mechanisms that produce alterations in cellular function. Roxithromycin, dirithromycin, and clindamycin were avidly concentrated by PMNs. Pentoxifylline and two derivatives (HWA-448 [torbafylline] and HWA-138 [albifylline]) increased the uptake of these antibiotics by PMNs, both in the resting state and during phagocytosis. Pentoxifylline, HWA-448, HWA-138, and the highly concentrated antibiotics each exerted an inhibitory effect on the stimulated respiratory burst response in PMNs. The combination of both pentoxifylline and a modulatory antibiotic (roxithromycin or clindamycin) inhibited superoxide production to a greater extent than either agent alone. This additive effect might be expected, since pentoxifylline and the modulatory antibiotics influence the respiratory burst activation pathway at different sites. The ability of pentoxifylline to augment the entry of antibiotics into neutrophils has important therapeutic implications. The consequences of this phenomenon might include improved intracellular bactericidal activity as well as efficient antibiotic delivery and release at sites of infection. PMID- 7492109 TI - In vitro activity of the semisynthetic glycopeptide amide MDL 63,246. AB - The in vitro activity of the semisynthetic glycopeptide amide MDL 63,246 against 293 U.S. clinical isolates of gram-positive cocci was determined by the broth microdilution method. When compared with teicoplanin, MDL 63,246 had improved activity against Staphylococcus epidermidis (MICs that inhibited 90% strains tested [MIC90s], 0.25 versus 8 micrograms/ml, respectively). Staphylococcus haemolyticus (MIC90s, 1 versus 32 micrograms/ml, respectively), and VanA Enterococcus faecium (MIC90s, 32 versus > or = 1,024 micrograms/ml, respectively). PMID- 7492108 TI - Antimicrobial activity of MDL 63,246, a new semisynthetic glycopeptide antibiotic. AB - MDL 63,246 is a semisynthetic derivative of the naturally occurring glycopeptide antibiotic MDL 62,476 (A40926). It was more active in vitro against Staphylococcus aureus and coagulase-negative staphylococci than MDL 62,476, teicoplanin, and vancomycin and was more active than mideplanin (MDL 62,873) against some isolates. MDL 63,246 had excellent activity against streptococci and teicoplanin-susceptible enterococci, and it also had in vitro activity against some VanA enterococcal isolates. It was more active than teicoplanin and vancomycin against acute staphylococcal, streptococcal, and enterococcal septicemia in immunocompetent and neutropenic mice. It was highly efficacious in reducing the bacterial load in the hearts of rats in staphylococcal endocarditis experiments and the bacterial load of Staphylococcus epidermis in a high infection model in neutropenic mice. The excellent in vivo activity of MDL 63,246 appears to correlate both with its in vitro antibacterial activity and with its long half-life in rodents. PMID- 7492110 TI - Comparative antimicrobial activities of the penem WY-49605 (SUN5555) against recent clinical isolates from five U.S. medical centers. AB - The in vitro activity of WY-49605 (SUN5555) (WY) was compared with those of cefaclor, cefixime, and amoxicillin-clavulanic acid against 2,958 consecutive clinical isolates from five medical centers and 402 respiratory pathogens from 18 other facilities. Most members of the family Enterobacteriaceae were inhibited by WY (MIC at which 50% of the isolates are inhibited [MIC50], < or = 2.0 micrograms/ml). MIC90s of > or = 8.0 micrograms/ml were observed for Enterobacter cloacae, Serratia spp., and Proteus mirabilis. WY was the most active drug against methicillin-susceptible Staphylococcus aureus (MIC90, 0.12 microgram/ml) and other coagulase-negative staphylococci (MIC90, 4.0 micrograms/ml). The four drugs were not active against nonenteric gram-negative bacilli, methicillin resistant Staphylococcus aureus, and Staphylococcus haemolyticus. At 2.0 micrograms/ml, WY inhibited 82% of Enterococcus faecalis strains and was equal to or superior to the other drugs against streptococci, Haemophilus influenzae, and Moraxella catarrhalis. PMID- 7492111 TI - Susceptibility of Acanthamoeba castellanii to contact lens disinfecting solutions. AB - A corneal isolate of Acanthamoeba castellanii was exposed to commercial contact lens disinfecting solutions containing hydrogen peroxide, benzalkonium chloride, polyaminopropyl biguanide, polyquaternium 1, and chlorhexidine-thimerosal. The minimum trophozoite amebicidal concentration and exposure times required to kill trophozoites and cysts were determined. Solutions containing hydrogen peroxide or chlorhexidine-thimerosal were active against both trophozoites and cysts. The benzalkonium chloride-based solution was effective only against trophozoites. Solutions containing polyaminopropyl biguanide or polyquaternium 1 were completely ineffective. The need for adequate exposure times must be stressed. PMID- 7492112 TI - Activity of penciclovir against Epstein-Barr virus. AB - Penciclovir inhibited the productive replication cycle of Epstein-Barr virus (EBV) in assays measuring infectious virus production, viral antigen expression, and viral DNA synthesis. In the test measuring inhibition of EBV DNA synthesis, 50% effective concentrations of penciclovir and acyclovir were 2.3 +/- 0.8 and 2.2 +/- 0.6 micrograms/ml, respectively. The 50% cell growth inhibitory concentration of penciclovir was > 100 micrograms/ml for both P3HR-1 and Raji cells. Penciclovir is a selective inhibitor of EBV in cell culture. PMID- 7492113 TI - Quinine plus clindamycin improves chemotherapy of severe malaria in children. AB - In a randomized trial, a 4-day quinine-clindamycin regimen was compared with the standard 7-day quinine regimen for 100 Gabonese children (50 children in each group) with severe Plasmodium falciparum malaria. In each group, only one patient died. Parasite clearance and fever clearance times were significantly shorter in the quinine-clindamycin group (P = 0.03 and P = 0.01, respectively) than in the quinine group, and significantly more recurring fever episodes occurred in the quinine group than in the quinine-clindamycin group shortly after initial fever clearance and parasite clearance (P < 0.001). PMID- 7492114 TI - Activities of three investigational fluoroquinolones (BAY y 3118, DU-6859a, and clinafloxacin) against Neisseria gonorrhoeae isolates with diminished susceptibilities to ciprofloxacin and ofloxacin. AB - Between 1 January 1992 and 31 December 1993, our laboratory, as part of the Gonococcal Isolate Surveillance Project, found that 39 of 673 isolates of Neisseria gonorrhoeae from one local sexually transmitted diseases clinic demonstrated decreased susceptibilities to both ciprofloxacin and ofloxacin. The MICs of BAY y 3118, DU-6859a, and clinafloxacin at which 90% of the gonococci with decreased susceptibility to ciprofloxacin and ofloxacin were inhibited were all 0.016 microgram/ml, which was eightfold higher than those for ciprofloxacin susceptible gonococci. Our report substantiates prior observations that diminished susceptibility to one quinolone is often associated with diminished susceptibility to other quinolones. PMID- 7492115 TI - Efficacy of the herbicide trifluralin against four P-glycoprotein-expressing strains of Leishmania. AB - Drug resistance has emerged as a major obstacle to chemotherapy for many infectious diseases. Trifluralin, an antimicrotubule herbicide, is a new experimental drug for treatment of leishmaniasis. Here, we found that it was effective against two strains of Leishmania that express the multidrug-resistant genes ldmdr1 and lmpgpA and two strains that express proteins that are immunologically cross-reactive with mammalian P glycoproteins. These results suggest that trifluralin is not subject to counteractions of these multidrug resistance mechanisms of Leishmania species. PMID- 7492116 TI - A novel integron-like element carrying the metallo-beta-lactamase gene blaIMP. AB - A plasmid-mediated metallo-beta-lactamase gene was cloned from a carbapenem resistant Serratia marcescens strain, AK9373. The metallo-beta-lactamase gene was identical to the blaIMP, and it was located in the space between an integrase like gene and an aac(6')-Ib-like gene. The deduced amino acid sequence for the putative integrase gene showed considerable identity (60.9%) to that of the Escherichia coli integrase reported. Sequences similar to the GTTRRRY and an atypical 59-base element containing a 67-bp inverted repeat sequence, which were peculiar to the integrase-dependent recombination, were also conserved in the flanking regions of the blaIMP gene. These findings imply that the metallo-beta lactamase gene in S. marcescens AK9373 is carried by a novel integron-like element that is mediated by a transferable large plasmid. PMID- 7492117 TI - Expression of Streptomyces peucetius genes for doxorubicin resistance and aklavinone 11-hydroxylase in Streptomyces galilaeus ATCC 31133 and production of a hybrid aclacinomycin. AB - The aklavinone 11-hydroxylase gene and two doxorubicin resistance genes cloned from Streptomyces peucetius subsp. caesius ATCC 27952 were introduced into doxorubicin-sensitive Streptomyces galilaeus ATCC 31133, an aclacinomycin producer. The doxorubicin resistance genes drrA and drrB endowed S. galilaeus with high-level resistance to doxorubicin, indicating that the resistance mechanism for doxorubicin might be different from that for aclacinomycin A. Transformation of S. galilaeus ATCC 31133 with plasmid pMC213 containing the aklavinone 11-hydroxylase gene (dnrF) resulted in the production of many red pigments. A new metabolite was purified, and the position of the newly introduced hydroxyl group was determined. This result indicated that the aklavinone 11 hydroxylase gene was stably expressed in S. galilaeus ATCC 31133 and that it gave rise to a hybrid aclacinomycin A which showed highly specific in vitro cytotoxicity against leukemia and melanoma cell lines. PMID- 7492118 TI - Salmonella typhimurium gyrA mutations associated with fluoroquinolone resistance. AB - Spontaneous quinolone-resistant mutants obtained from Salmonella typhimurium Su694 were screened for mutations by direct DNA sequencing of an amplified PCR gyrA fragment. Substitutions Ser-83-->Phe (Ser83Phe), Ser83Tyr, Asp87Tyr, and Asp87Asn and double mutation Ala67Pro-Gly81Ser, which resulted in decreased sensitivities to ciprofloxacin, enoxacin, pefloxacin, norfloxacin, ofloxacin, and nalidixic acid, were found. The levels of resistance to quinolones for each mutant were determined. PMID- 7492119 TI - Analysis of mutations at position 184 in reverse transcriptase of human immunodeficiency virus type 1. AB - We have analyzed recombinant human immunodeficiency virus type 1 reverse transcriptases that contain mutations at position 184. These variants retain high levels of RNA-dependent DNA polymerase activity and show resistance to ddITP. However, the mutants varied in their ability to polymerize processively. The variants Met184Ile and Met184Val showed slight reductions in processivity relative to that of the wild-type enzyme; the variants Met184Ala and Met184Leu showed considerable reductions in their processivity. PMID- 7492120 TI - The renal membrane dipeptidase (dehydropeptidase I) inhibitor, cilastatin, inhibits the bacterial metallo-beta-lactamase enzyme CphA. AB - The Aeromonas hydrophila AE036 chromosome contains a cphA gene encoding a metallo beta-lactamase which is highly active against carbapenem antibiotics such as imipenem. Here we show that the cphA gene product shares inhibitory similarities with a mammalian zinc peptidase, membrane dipeptidase (MDP; dehydropeptidase I). Both enzymes are able to hydrolyze imipenem and are inhibited by cilastatin. The active site similarities of these enzymes are not reflected in any significant primary sequence similarity. PMID- 7492121 TI - Susceptibilities of several drug-resistant herpes simplex virus type 1 strains to alternative antiviral compounds. AB - Resistant herpes simplex virus type 1 strains were obtained under the selective pressure of acyclovir, ganciclovir, bromovinyldeoxyuridine, foscarnet, 2 phosphonylmethoxyehtyl (PME) derivatives of adenine and 2,6-diaminopurine, 3 hydroxy-2-phosphonylmethoxypropyl derivatives of adenine and cytosine, and 2 amino-7-(1,3-dihydroxy-2-propoxymethyl)purine (S2242). The drug susceptibility profiles of resistant strains point to differences in the modes of action of PME and 3-hydroxy-2-phosphonylmethoxypropyl derivatives and common mechanisms of action of foscarnet, S2242, and PME derivatives against herpes simplex virus type 1 replication. PMID- 7492122 TI - In vitro evaluation of DV-7751a, a new fluoroquinolone with an enhanced spectrum of activity against gram-positive aerobic organisms and anaerobes. AB - DV-7751a is an investigational fluoroquinolone with improved spectrum and potency against gram-positive bacteria. We studied the in vitro activity of this compound against 771 recent clinical isolates by the reference agar and broth microdilution methods. Its activity was compared with those of ciprofloxacin, levofloxacin, and ofloxacin and with those of ceftazidime, oxacillin, and gentamicin where relevant. DV-7751a was four- to eightfold more active than the comparison fluoroquinolones against Enterococcus spp. (including vancomycin resistant strains), Staphylococcus spp. (including oxacillin-resistant Staphylococcus aureus), Streptococcus spp., other gram-positive pathogens, and some anaerobes. The DV-7751a activity against most gram-negative species was similar to that of ofloxacin and ciprofloxacin. DV-7751a appears to be a candidate for the therapy of some mixed-flora infections and the treatment of bacteria resistant to the current fluoroquinolones. PMID- 7492123 TI - Prospective randomized trial of efficacy of ganciclovir versus that of anti cytomegalovirus (CMV) immunoglobulin to prevent CMV disease in CMV-seropositive heart transplant recipients treated with OKT3. AB - We compared the efficacy of ganciclovir versus that of cytomegalovirus (CMV) immunoglobulin for the prevention of CMV disease in 31 CMV-seropositive heart transplant recipients who had received early immunoprophylaxis with OKT3 monoclonal antibodies. The incidence of CMV disease and visceral involvement was much higher in the CMV immunoglobulin group than in the ganciclovir group (40 versus 6%, respectively; P = 0.03). No adverse effects were found in the CMV immunoglobulin group, but 19% of the patients in the ganciclovir group developed mild leukopenia or a mild increase in their serum creatinine levels. PMID- 7492124 TI - Improvement in androgenetic alopecia (stage V) using topical minoxidil in a retinoid vehicle and oral finasteride. PMID- 7492125 TI - Dermatomyositis without muscle weakness. Long-term follow-up of 12 patients without systemic corticosteroids. AB - BACKGROUND AND DESIGN: We retrospectively studied 12 patients with dermatomyositis (DM) with at least one laboratory criterion of muscle involvement but without muscle weakness. We reported their outcome without systemic corticosteroid treatment and searched for distinctive baseline data and associated neoplasia among a group of 29 patients with classic DM. RESULTS: After a mean follow-up of 5.3 years, complete or partial skin improvement occurred in nine patients with DM without muscle weakness, and muscle enzyme levels were normalized in all. When compared with the classic DM group, no statistical differences were found for associated neoplasia or for any baseline data studied except, as expected, for muscle parameters. CONCLUSIONS: The lack of distinctive baseline data between DM without muscle weakness and classic DM suggests that there is a continuum from amyopathic to classic DM. However, DM without muscle weakness is a clinical entity with respect to favorable long-term prognosis. Unless frank muscle weakness develops, treatment with systemic corticosteroids can be avoided. PMID- 7492127 TI - Positron emission tomography and ultrasonography. A comparative retrospective study assessing the diagnostic validity in lymph node metastases of malignant melanoma. AB - BACKGROUND AND DESIGN: A retrospective study involving 20 patients with melanoma with clinically suspicious lymph nodes was conducted to compare the diagnostic validity of fludeoxyglucose F 18 positron emission tomography (PET) and real-time ultrasonography in lymph node metastases of malignant melanoma. RESULTS: A total of 83 lymph nodes were assessed with ultrasonography and PET. Imaging results were confirmed by histologic studies or close follow-up ultrasonographic examinations. Positron emission tomography revealed a sensitivity of 74% and a specificity of 93%. Both investigative methods show comparative sensitivity and specificity. CONCLUSIONS: Ultrasonography is much easier to perform, less time consuming, and less expensive than PET and it is nonhazardous; therefore, it is ideal for follow-up procedures. Since in routine staging procedures, only sites of expected lymph node involvement are examined, there is a risk of metastases being missed in cases of atypical drainage patterns. Fludeoxyglucose F 18 PET can image proliferating tumors in multiple organ systems and lymph node sites in one session, making it suitable for screening in primary staging procedures and for monitoring response to therapy. Since it is based on metabolic changes, there is good differentiation between scar and tumor tissue. Major disadvantages are restricted access to investigation centers, high imaging costs, and limited anatomical location of metastatic lesions. We conclude that PET does not offer significant advantages in the diagnosis of lymph node metastases compared with ultrasonography. PMID- 7492126 TI - Membrane attack complex deposits in cutaneous lesions of dermatomyositis. AB - BACKGROUND AND DESIGN: The presence of membrane attack complex of complement (MAC) deposits in the intramuscular vasculature of biopsy specimens taken from patients with dermatomyositis (DM) has been implicated in the pathogenesis of this myopathy. The purpose of this study was to investigate the presence of MAC deposition in the skin lesions of patients with DM. Using immunohistochemical methods, we examined 22 biopsy specimens from lesional skin, six biopsy specimens from uninvolved skin, and 12 muscle biopsy specimens from patients with DM for the presence of MAC and vitronectin and CD59, two regulatory proteins of complement. RESULTS: The deposition of MAC was demonstrated in a large percentage of biopsy specimens obtained from the lesional skin of patients with DM. Deposits were found along the dermoepidermal junction in 19 (86%) of 22 biopsy specimens. Deposits on the vessel walls of the dermis were found in 17 (77%) of 22 biopsy specimens; but only in six of these biopsy specimens (27%) were deposits present in more than 10% of blood vessels. In contrast, deposits along the dermoepidermal junction and the vessel walls of the dermis were absent in specimens from uninvolved skin. In 12 muscle biopsy specimens obtained simultaneously from these patients, MAC deposits were found on the vessel walls in nine (75%), but only in six (50%) were deposits found in more than 10% of the intramuscular vessels. The pattern of vitronectin immunoreactivity in skin and muscle biopsy specimens obtained from patients with DM was similar to MAC deposits. The expression of CD59 was normal in all skin and muscle biopsy specimens. CONCLUSIONS: The deposition of MAC was found in a high percentage of biopsy specimens from the lesional skin of patients with DM; it was absent in uninvolved skin. These findings suggest that the complement system may be involved in the pathogenesis of the skin lesions of DM. PMID- 7492128 TI - Colchicine in the treatment of cutaneous leukocytoclastic vasculitis. Results of a prospective, randomized controlled trial. AB - BACKGROUND AND DESIGN: Cutaneous leukocytoclastic vasculitis is an inflammatory vascular disease with a variable course. There is no defined therapy for this entity. Contradictory data on the effect of colchicine have been reported. To determine the efficacy of colchicine in cutaneous leukocytoclastic vasculitis, 41 patients were randomly selected to receive oral colchicine, 0.5 mg twice daily, or topical emollients. Response to treatment was judged according to the reduction in the number of lesions. After 1 month, in those patients in whom a complete or no response was achieved, therapy was withdrawn; in those with a partial response, treatment was maintained for the following 2 months. At the end of 3 months, treatment was continued only in those patients in whom a relapse occurred. RESULTS: Twenty patients in each group completed 1 month of treatment. One patient taking colchicine dropped out because of diarrhea. At the end of the first month of the study, five patients in the control group and four in the colchicine group achieved a complete response. Nine patients who had a partial response (four in the colchicine group and five in the control group) continued to receive treatment for the following 2 months. Three patients in the colchicine group suffered a relapse after discontinuing therapy but experienced remission with reinstitution of therapy. At the end of the 3 month period, 12 patients in the colchicine group and 10 patients in the control group showed no significant response. Complete response was achieved in five patients in the colchicine group and in seven in the control group. At the 1-year follow-up, 10 patients in each group had no clinical evidence of cutaneous vasculitis. CONCLUSIONS: Colchicine had no significant therapeutic effect in this controlled study. However, the finding that relapse occurred on cessation of colchicine therapy in three complete responders suggests that colchicine can be effective in some patients, despite our negative results. PMID- 7492129 TI - The antipruritic effect of 5% doxepin cream in patients with eczematous dermatitis. Doxepin Study Group. AB - BACKGROUND AND DESIGN: Eczematous dermatitis is commonly characterized by intense pruritus. Current treatment modalities for this condition, regardless of its cause, are primarily directed at blunting the cutaneous inflammatory response and thereby providing relief of pruritus. To expand on our previous findings in atopic dermatitis, the present multicenter double-blind trial was conducted to evaluate the safety and antipruritic efficacy of 5% doxepin hydrochloride cream in patients with lichen simplex chronicus (n = 136), nummular eczema (n = 87), or contact dermatitis (n = 86). A total of 309 patients with moderate to severe pruritus were randomly assigned to apply either doxepin cream (n = 154) or vehicle cream (n = 155) to eczematous areas four times per day for a period of 7 days. Efficacy was assessed using a pruritus severity rating scale, a Physician's Global Evaluation for pruritus relief, and a Visual Analogue Scale for pruritus relief. RESULTS: Twenty-four hours after initiation of treatment, and continuing throughout the remainder of the study, patients treated with doxepin cream experienced significantly greater pruritus relief than did vehicle-treated patients as determined by all efficacy parameters (P < .002). Sixty percent of doxepin-treated patients experienced pruritus relief within 24 hours. The response rate increased to 84% by conclusion of the study. As judged by significant changes (P < or = .05) occurring in at least one assessment of efficacy, doxepin cream provided pruritus relief in all forms of eczematous dermatitis that were examined. The study medication was well tolerated. The two most common adverse effects, stinging at the site of application and drowsiness, were usually transient and mild to moderate in severity. CONCLUSION: Topical application of doxepin provides significant antipruritic activity with a favorable safety profile, suggesting a role for doxepin cream in the symptomatic treatment of pruritus associated with eczematous dermatitis. PMID- 7492130 TI - Localized chrysiasis induced by laser therapy. AB - BACKGROUND: Chrysiasis is a rare blue-gray skin discoloration that occurs in sun exposed sites of some patients who receive gold salts. A unique case of localized chrysiasis developed immediately after Q-switched ruby laser (694 nm) irradiation for postinflammatory hyperpigmentation secondary to granuloma faciale in a patient who was receiving long-term gold sodium thiomalate therapy for psoriatic arthritis. Skin biopsy specimens showed striking changes in the ultrastructural characteristics of cutaneous gold deposits following laser treatment. OBSERVATIONS: A blue-gray skin discoloration developed immediately after laser exposure and persisted unchanged after 1 year. Transmission electron microscopy of skin biopsy specimens showed electron-dense gold deposits. Before laser irradiation, these deposits were 106 +/- 35 (mean +/- SD) nm in diameter and faceted, consistent with a crystalline structure. Posttreatment deposits were round, smaller, measured 16 +/- 4 nm, and resembled colloidal gold. Identical findings were observed in an area of sun-protected skin treated with the Q switched ruby laser; irradiation with a pulsed dye laser at 585 nm had no effect. CONCLUSIONS: Localized chrysiasis was induced in a patient receiving parenteral gold therapy who underwent treatment with a Q-switched ruby laser. This form of chrysiasis resulted from a structural alteration in dermal gold deposits. A similar physiochemical modification in gold deposits induced by UV light may explain the localization of chrysiasis to sun-exposed skin in affected patients. PMID- 7492132 TI - Persistent subcutaneous nodules in patients hyposensitized with aluminum containing allergen extracts. AB - BACKGROUND: The development of persistent nodules that cause pain and itching at a vaccination or hyposensitization injection site is a rare event. These lesions have been mainly attributed to a hypersensitivity reaction to aluminum hydroxide, which is used as an absorbing agent in many vaccines and hyposensitization preparations. Patch tests with standard antigens and aluminum compounds and histopathologic and ultrastructural studies were performed on 10 patients with persistent subcutaneous nodules on the upper part of their arms after injection of aluminum-adsorbed dust and/or pollen extracts. OBSERVATIONS: The nodules appeared 1 month to 6.5 years after injections. The results of patch tests with 2% aluminum chloride were positive in five patients. Histopathologic examination revealed two different patterns: some biopsy specimens (from lesions of less than 9 months' duration) showed a pure foreign body histiocytic reaction characterized by extracellular amorphous dermal basophilic deposits with a histiocytic macrophagic reaction; others showed a delayed hypersensitivity granulomatous reaction in association with an histiocytic foreign body response. The lesions were characterized by a unifocal or multifocal unencapsulated granulomatous reaction in the deep dermis and/or subcutaneous tissue. Eosinophilic necrotic areas surrounded by dense fibrous bands and a massive inflammatory infiltrate (lymphoid follicles, large histiocytic cells, abundant eosinophils, and some plasma cells) were observed. A granular basophilic material in extracellular spaces and within the cytoplasm of some histiocytes was also noted. Electron microscopic studies revealed intracytoplasmic and extracellular deposits of a fibrillar electron-dense material. CONCLUSIONS: Persistent subcutaneous nodules that develop after the administration of aluminum-containing preparations may show two characteristic histopathologic patterns. A pure histiocytic foreign body reaction was observed in early lesions, and a delayed hypersensitivity granulomatous reaction was seen in older lesions. No relationship between histopathologic pattern and patch test results was observed. Aluminum-free preparations should be used in patients in whom these nodules develop. PMID- 7492131 TI - Treatment of cafe au lait macules with lasers. A clinicopathologic correlation. AB - BACKGROUND: Cafe au lait macules (CALMs) respond variably to treatment with different lasers. This study was done to determine whether the type of laser and the individual histologic features of the CALMs could predict clinical response to treatment. Nine CALMs were treated with both the frequency-doubled Q-switched neodymium: YAG laser (wavelength, 532 nm; beam diameter, 2.0 mm) and the Q switched ruby laser (wavelength, 694 nm; beam diameter, 5.0 mm). Both lasers were used at a fluence of 6.0 J/cm2. Biopsy specimens of the CALMs were obtained before and after treatment. Clinical follow-up was done at 1-, 3-, and 6-month intervals. OBSERVATIONS: Both lasers yielded variable responses to treatment. Two histologic subtypes of CALMs were identified, but these different subtypes did not predict clinical outcome after laser treatment. CONCLUSIONS: Cafe au lait macules respond variably to treatment with both the Q-switched ruby laser and the frequency-doubled Q-switched neodymium:YAG laser. Further research might address the effect of using multiple treatments. In view of these results, clinicians using lasers to treat CALMs should inform their patients of the potential for recurrence or darkening of CALMs. PMID- 7492133 TI - 'Membranous aplasia cutis' with hair collars. Congenital absence of skin or neuroectodermal defect? AB - BACKGROUND: The skin and the nervous system are both derived from ectoderm. Separation of neural ectoderm from epithelial ectoderm occurs concurrently with the closure of the neural tube. This chronologic association may explain the cutaneous abnormalities often found overlying neural tube defects. A ring of dark long hair encircling a congenital scalp lesion (the hair collar sign) is one such marker and is often associated with encephaloceles, meningoceles, and heterotopic brain tissue. OBSERVATIONS: We describe six children with aplasia cutis who displayed the hair collar sign. Aplasia cutis is a relatively heterogeneous disorder; however, these lesions had a unique and strikingly similar appearance. This subtype of aplasia cutis, which we have termed membranous aplasia cutis, shares several clinical and histologic features with cranial neural tube defects. CONCLUSIONS: We propose that membranous aplasia cutis is a form fruste of a neural tube defect and may be derived from a similar embryological defect. Recent advances in the understanding of cranial neural tube closure may provide support for this hypothesis. PMID- 7492136 TI - A bullous eruption in an elderly woman. Vancomycin-associated linear IgA dermatosis (LAD). PMID- 7492134 TI - Linear IgA bullous dermatosis. Characterization of a subset of patients with concurrent IgA and IgG anti-basement membrane autoantibodies. AB - BACKGROUND AND DESIGN: The term linear IgA bullous dermatosis defines an immune mediated blistering skin disease characterized by pruritic blisters, subepidermal separation with neutrophilic infiltration, and linear IgA antibody deposition at the basement membrane zone (BMZ). However, some patients with linear IgA bullous dermatosis demonstrate both IgA and IgG anti-BMZ autoantibodies on immunofluorescence. We describe four such patients and attempt to define this group of patients by studying their clinical, histopathologic, immunopathologic, immunoultrastructural, and immunochemical characteristics. RESULTS: Clinically, all four patients had a generalized pruritic blistering skin disease consistent with linear IgA bullous dermatosis. Histopathologically, all four cases demonstrated subepidermal blister formation with a predominantly neutrophilic dermal infiltrate. Immunopathologically, tissue-bound IgA (in four of four patients) and IgG (in three of four patients) antibodies were detected at the BMZ. Circulating IgA (in four of four patients) and IgG (in four of four patients) labeled the epidermal BMZ of normal human skin fractured at the lamina lucida and did not label the dermal BMZ. By immunoblot analyses, IgA (in three of three patients) and IgG (in one of three patients) circulating antibodies labeled the 97-kd linear IgA bullous dermatosis antigen. By direct immunoelectron microscopy, IgA and IgG antibodies were localized (in two of two patients) exclusively within the upper lamina lucida of the BMZ. CONCLUSIONS: Four cases of an immune-mediated blistering skin disease typical for linear IgA bullous dermatosis demonstrated both IgA and IgG anti-BMZ autoantibodies against the linear IgA bullous dermatosis antigen within the upper lamina lucida. We conclude that linear IgA bullous dermatosis should include a subgroup of patients with both IgA and IgG anti-BMZ autoantibodies. PMID- 7492135 TI - A child with antibodies targeting both linear IgA bullous dermatosis and bullous pemphigoid antigens. AB - BACKGROUND: Some patients with subepidermal blistering diseases show clinical, histologic, and immunopathologic features of both linear IgA bullous dermatosis and bullous pemphigoid. Such patients can be further characterized by defining the target of their circulating autoantibodies. We present the first case report of a child with linear deposits of IgA and IgG with circulating autoantibodies characteristic of both linear IgA bullous dermatosis and bullous pemphigoid. OBSERVATIONS: Widely distributed subepidermal vesicles showing neutrophils in the dermal papillae developed in a 3-year-old boy. Direct immunofluorescence microscopy of perilesional skin revealed linear deposits of IgA, IgG, and C3 in the epidermal basement membrane. The patient responded to therapy with dapsone, and after 6 months, it was possible to discontinue treatment. Circulating IgA antibodies from this child bound the epidermal side of 1-mol/L saline-split skin and immunoblotted the 97-kd linear IgA bullous dermatosis antigen. Circulating IgG antibodies bound the epidermal and, at low titer, dermal sides of split skin. These IgG antibodies immunoblotted and immunoprecipitated bullous pemphigoid antigens 1 and 2. CONCLUSIONS: Linear deposits of IgA and IgG in the epidermal basement membrane of patients with subepidermal bullous lesions may signify the coexistence of circulating autoantibodies directed against linear IgA bullous dermatosis and bullous pemphigoid antigens. PMID- 7492137 TI - Hyperpigmented patches on the dorsa of the feet. Minocycline pigmentation. PMID- 7492138 TI - Bilateral jawline nodules in a child with a brain-stem glioma. Poststeroid panniculitis. PMID- 7492139 TI - A penile ulceration in a patient with the acquired immunodeficiency syndrome. Foscarnet-induced penile ulceration. PMID- 7492140 TI - Melasma. Etiologic and therapeutic considerations. AB - BACKGROUND: Melasma is a common acquired symmetric hypermelanosis characterized by irregular light- to gray-brown macules and patches involving sun-exposed areas of skin. Etiologic factors in the pathogenesis of melasma include genetic influences, exposure to UV radiation, pregnancy, hormonal therapies, cosmetics, phototoxic drugs, and antiseizure medications. OBSERVATIONS: Melasma is often a therapeutically challenging disease, and current treatments include hypopigmenting agents, chemical peels, and lasers. Hypopigmenting agents include phenolic and nonphenolic derivatives. Phenolic agents include hydroquinone and hydroquinone combination preparations. Despite controversies regarding the issue of hydroquinone-induced ochronosis, hydroquinone remains the most effective topically applied bleaching agent approved by the Food and Drug Administration for the treatment of melasma. Nonphenolic bleaching agents include tretinoin and azelaic acid. Superficial, medium, and deep chemical peels are more often used in lighter-complexioned patients. Such peels should be used with caution in blacks. Although lasers have demonstrated significant efficacy in the treatment of a variety of hyperpigmentary disorders, their precise efficacy and place in the therapy of melasma have yet to be established. CONCLUSIONS: In the hierarchy of therapies for melasma, the treating physician must consider the devastating psychosocial impact of pigmentary imperfections within the realm of the benefits and risks associated with each treatment. PMID- 7492141 TI - Amyopathic dermatomyositis. PMID- 7492142 TI - Tuberous sclerosis. The persistent challenge of clinical diagnosis. PMID- 7492143 TI - Langerhan's cells in skin tumors. PMID- 7492144 TI - How high the SPF? PMID- 7492145 TI - Trichogenic tumors. PMID- 7492146 TI - Unsolicited mailings to dermatologists. PMID- 7492147 TI - More on the frequency of segmental neurofibromatosis. PMID- 7492148 TI - The significance of a single periungual fibroma: report of seven cases. PMID- 7492149 TI - Budesonide patch testing. PMID- 7492150 TI - Toxic reaction from topically applied doxepin in a child with eczema. PMID- 7492151 TI - Melanoma in situ of the sole that extended into a skin graft: reflections on the biology of melanoma. PMID- 7492152 TI - Molecular biology of coeliac disease. PMID- 7492153 TI - Is community screening for amblyopia possible, or appropriate? AB - Photoscreeners are becoming increasingly available and are being widely used to screen for visual abnormalities in young children. However, consideration of accepted criteria for screening programs indicates there is still much further research that needs to be carried out before amblyopia screening could be recommended as a routine component of a community health surveillance program--an adequate description of the potential consequences of an individual developing amblyopia has yet to be provided and the natural history of the condition and factors that determine the effectiveness of treatment have yet to be fully described. While there is the promise of technology that satisfies specific test requirements, this still needs to be trialed in community settings and community trials are required before it will be possible to determine whether the costs that will be incurred in carrying out routine screening and in providing the resources for treatment are warranted. In conclusion, the development of new and possibly more effective technology for screening is only part of the answer to the amblyopia question. While superficially this technology makes screening for amblyopia a possibility, we do not know at this stage whether or not it is appropriate. PMID- 7492154 TI - Long-term survival and state of health after paediatric intensive care. AB - Survival and state of health were assessed one year after discharge in 468 children admitted to an intensive care unit (ICU). ICU mortality was 7.5%, cumulative hospital mortality 8.3%, and one year mortality 10.5%. An established six domain health status classification was used, comprising sensation, mobility, emotion, cognition, self care, and pain to determine the presence, type, and severity of chronic health impairment. This classification has not been validated for infants, who were therefore excluded. After one year, of the 254 patients eligible for analysis, 80 (31.5%) had no overall health impairment (no affected domains) preceding admission; of these, 11 had died in ICU and 69 were long-term survivors, among whom 45 recovered to perfect health. There was overall health impairment (> or = 1 affected domain) preceding admission in 174 of 254 patients (68.5%). However, after one year, overall state of health was improved or equal to the preadmission state in 164 of 226 survivors (72.6%). In domain-specific health, the proportion improving or remaining unchanged varied from 77.9% (emotional functions) to 89.4% (mobility and pain). Consequently, despite the large number with health impairment before admission, cumulative one year survival was favourable and health status in three quarters of the population was preserved. PMID- 7492155 TI - Controlled trial of a few foods diet in severe atopic dermatitis. AB - Eighty five children (median age 2.3 years, range 0.3 to 13.3 years) with refractory atopic dermatitis affecting more than 12% of the body surface area, were randomly allocated to receive a few foods diet (eliminating all but five to eight foods) supplemented with either a whey hydrolysate (n = 27) or a casein hydrolysate formula (n = 32), or to remain on their usual diet and act as controls (n = 26), for a six week period. Thirty five patients who received the diet and four controls had to be withdrawn because of non-compliance with the diet or intercurrent illness. The change in dermatitis severity was evaluated by a blinded observer who estimated the extent and severity of the dermatitis, using a skin severity score. After six weeks, there was a significant reduction in all three groups in the percentage of surface area involved (controls, median reduction (MR) = 4.9% (95% confidence interval 1.5%, 11.9%); whey hydrolysate group, MR = 17.8% (8.3%, 23.0%); casein hydrolysate group, MR = 5% (1.6%, 21.2%), and skin severity score (controls, MR = 15.9 (5.0, 22.5); whey hydrolysate group, MR = 21.8 (12.8, 30.2); casein hydrolysate group, MR = 13.5 (3.4, 38.0). Sixteen (73%) of the 22 controls and 15 (58%) of the 24 who received the diet showed a greater than 20% improvement in the skin severity score. This study failed to show benefit from a few foods diet. PMID- 7492156 TI - Nutritional content of few foods diet in atopic dermatitis. AB - The nutritional content of a few foods diet, supplemented with a casein hydrolysate formula (n = 24) or a whey hydrolysate formula (n = 21), was studied in 45 children with atopic dermatitis. The six day weighed food inventory record method was used to estimate the mean daily intake of energy, protein, calcium, iron, zinc, folate, and vitamin C on normal diet and on the few foods diet. The diet was associated with a significant reduction in protein and calcium intake in both groups, and in energy intake in the casein hydrolysate group. The median daily volume of hydrolysate milk taken was 10.5 ml/day (range 0-840 ml/day) for the casein hydrolysate group and 267 ml/day (range 0-1300 ml/day) for the whey hydrolysate group. Whey hydrolysate appears to be more palatable than casein hydrolysate, which is a potential advantage in the maintenance of an adequate intake in children on a few foods diet. PMID- 7492157 TI - Comparison of an elemental with a hydrolysed whey formula in intolerance to cows' milk. AB - In a double blind study, 40 infants with cows' milk intolerance of various causes were randomised to receive a nutritionally complete formula in which nitrogen was supplied either as whey hydrolysate or amino acids. The median age of infants was 10 weeks (range 36 weeks' gestation to 108 weeks' postnatal age). After a median follow up period of 25 weeks there was no significant difference in dietary intake between the formulas. Twenty four weeks after entry, weight and weight for length improved equally on both formulas. Plasma albumin improved significantly on the hydrolysed whey formula but not in the amino acid group. Both milks were palatable and normal intakes of formula were maintained. Biochemical and haematological indices remained within normal limits. There was no difference in stool frequency and vomiting between the two formulas. Two infants developed a probable allergic colitis while receiving hydrolysed whey. Amino acid formula may have a role in the management of atopic infants with severe cows' milk intolerance who have already reacted to whey or casein hydrolysate formula. PMID- 7492158 TI - The effect of iron in formula milk after 6 months of age. AB - Ninety two normal birthweight infants aged 6 months entered a double blind controlled trial which compared a follow on formula milk with no added iron against the same formula milk containing 1.2 mg of iron per 100 ml. There was no significant difference in the social class or demographic characteristics of the two treatment groups or in the proportion of each group completing the trial. There was no difference between the two groups in the quantity of milk taken but the amounts taken lessened between 6 and 18 months of age. There was no difference between the two groups with respect to mean haemoglobin and median serum ferritin at 6, 9, 12, 15, and 18 months of age. Very few infants developed iron deficiency anaemia in either group but there was a tendency for serum ferritin levels to fall between 6 and 18 months of age in both groups. The results suggest that iron added to follow on milk was not an important source of dietary iron in the infants studied. PMID- 7492159 TI - Persistent increase in plasma and urinary leukotrienes after acute asthma. AB - Leukotrienes may mediate bronchoconstriction in asthma. Cysteinyl leukotriene production rises in vivo after allergen challenge, but few reports describe leukotriene concentrations in clinical asthma or in children. Using high performance liquid chromatography/radioimmunoassay, plasma and urinary leukotrienes in asthmatic children (aged 5-10 years) were measured during an acute exacerbation (peak expiratory flow (PEF) < 65%, n = 10) and one month later (PEF 74-169%, n = 9), and in non-atopic normal children (aged 1.3-13.2 years). In the asthmatics, geometric mean (95% confidence interval) plasma leukotriene B4 (LTB4) was 746 pg/ml (398 to 1403) acutely and 1026 pg/ml (662 to 1593) in remission, compared with 369 pg/ml (167 to 728) in the normal children (n = 14). Plasma cysteinyl leukotrienes were low or undetectable, but urinary leukotriene E4 (LTE4) was higher in the asthmatics during an acute episode (210 pmol/mmol creatinine, 101 to 454) and at follow up (179 pmol/mmol, 110 to 293), compared with the normal children (98 pmol/mmol, 81 to 118, n = 41). This persistent increase in plasma LTB4 and urinary LTE4 concentrations one month after a severe asthmatic episode suggests leukotriene production is related to chronic inflammation rather than to acute bronchoconstriction. PMID- 7492160 TI - A non-electrostatic spacer for aerosol delivery. AB - A pear shaped non-electrostatic spacer, composed of steel with a volume of 250 ml and equipped with a facemask containing integrated inlet and outlet valves for inspiration and expiration, was compared with three plastic spacers. The plastic spacers were primed with repeated puffs from a budesonide pressurised metered dose inhaler (p-MDI) to minimise the electrostatic charge on the plastic. The procedure prolonged the half life (t1/2) of the aerosol in the Nebuhaler from nine to 32 seconds. A normal cleaning procedure reduced the aerosol t1/2 back to baseline. The t1/2 of the aerosol in the metal spacer was 27 seconds and independent of the use of p-MDI. In vitro the maximum dose of budesonide from a p MDI, expressed as a percentage of the nominal dose, was 56% from the non electrostatic spacer, 61% from the Nebuhaler, 45% from the Babyhaler, and 30% from the AeroChamber. In 124 children, age 6 months to 6 years, suspected to have asthma the non-electrostatic spacer delivered a mean total dose of budesonide aerosol of 39% of the nominal dose, which was significantly higher than the Babyhaler (28%), the Nebuhaler (21%), and the AeroChamber (19%). These differences were most pronounced in children younger than 4 years. The improved dose delivery from the small volume non-electrostatic spacer is probably related to the non-electrostatic spacer material and the valves which assured unidirectional airflow from the spacer without adding any dead space in the inspiratory channel. The non-electro-static spacer should improve the cost effectiveness of aerosol treatment and, as the counteracting effects of proming and recharging of the plastic from cleaning are avoided, should deliver a more reliable dose. PMID- 7492161 TI - Birth weight at term and lung function in adolescence: no evidence for a programmed effect. AB - It has been suggested that factors which influence low birth weight at term may be associated with reduced lung function in later life. This hypothesis was investigated in a comparative (retrospective) cohort study of 164 matched pairs of subjects where the observers responsible for tracing and studying the subjects were unaware of their case or control status. The subjects, born in Cardiff between 1975 and 1977, were of mean age 15.7 years. Cases (low birth weight (< 2500 g) at term) were matched with controls (normal birth weight (3000-3800 g) at term) for sex, parity, place of birth, date of birth, and gestation. Lung function was measured using a portable spirometer. The corrected mean differences (95% confidence interval) in forced vital capacity (FVC) and flow when 50% or 25% of the FVC remains in the lungs between the cases and controls were respectively 41 ml (-140 to 58), -82 ml/sec (-286 to 122), and -83 ml/sec (-250 to 83). None of these differences were statistically significant. These results are inconsistent with the hypothesis that low birth weight at term is associated with reduced lung function in adolescence. PMID- 7492163 TI - Prevalence and correlates of obesity in insulin dependent diabetic patients. AB - The prevalence of obesity, according to sex and pubertal stage, and the correlations between obesity and metabolic data were investigated in 286 diabetic patients (164 boys, 122 girls) with mean (SD) age 15.3 (3.2) years and mean (SD) duration of diabetes 7.5 (4.1) years. Prevalence of obesity according to the body mass index (BMI) criteria was 6.3%. Girls were more often obese than boys but the prevalence approached statistical significance only for the BMI criteria, at 9.8% v 3.7% (chi 2 = 3.5; p = 0.06); obesity was independent of pubertal stage. Distribution of BMI values of diabetic girls was skewed towards the high centiles of the INSERM tables: < 25th centile, 8.6%; 25th-50th centile, 17.3%; 50th-75th centile, 25.9%; > 75th centile, 48.2% (chi 2 = 19.17, p < 0.0005). BMI values of diabetic boys were homogeneously distributed. Age, duration of diabetes, insulin requirement, daily number of insulin injections, and metabolic control (HbA1c) were comparable in obese and non-obese diabetic patients. Moreover metabolic control and insulin requirements were comparable between diabetic patients with BMI above and below the 50th centile of the INSERM tables after matching for sex. In conclusion the prevalence of obesity in diabetic children and adolescents is quite similar to the prevalence reported in their non-diabetic peers. Obesity and BMI excess correlate with female gender but are independent of insulin requirement and metabolic control. PMID- 7492164 TI - Randomised comparison of ondansetron and metoclopramide plus dexamethasone for chemotherapy induced emesis. AB - The serotonin (5HT3) antagonist ondansetron was compared in a randomised study with metoclopramide and dexamethasone for the prevention of chemotherapy induced emesis. Thirty children aged 1-15 years with acute lymphoblastic leukaemia received 'intensification modules' according to the MRC United Kingdom acute lymphoblastic leukaemia regimen UKALL XI. This contains the moderately emetogenic drugs daunorubicin, etoposide, and cytarabine. Fifteen children received an intravenous loading dose of ondansetron followed by intravenous or oral doses 12 hourly for five days. Fifteen children received intravenous metoclopramide every six hours for three days with a loading dose of dexamethasone, repeated every eight hours for three days intravenously or orally. Efficacy was assessed by a diary card documenting the incidence of nausea, retching, or vomiting. In the 24 hour period after starting chemotherapy, ondansetron was more effective, with a complete or major response rate of 93%, compared with 33% using metoclopramide/dexamethasone. PMID- 7492162 TI - Effect of general anaesthesia on prolonged intraoesophageal pH monitoring. AB - Investigation of gastro-oesophageal reflux often includes endoscopy, usually under general anaesthesia, and pH monitoring. In most cases, the pH probe is passed when the child is awake and is poorly tolerated. The effect of general anaesthesia on pH monitoring is unknown. The aim of the study was to determine if placing the probe in the anaesthetised child gives a representative pH study. Twenty children aged 4 months to 13 years underwent oesophago-gastroduodenoscopy under general anaesthesia. A pH electrode was placed under direct vision in the distal oesophagus. pH monitoring was begun after completion of anaesthesia and continued for 18-24 hours. The study was repeated within 14 days without anaesthetic. The reproducibility of values of percent pH < 4, number of reflux episodes/hour, reflux episodes lasting > 5 min, and longest reflux episode was 85%, 90%, 75%, and 75% respectively. These results are comparable with those in adults and children in whom pH studies were performed on consecutive days (without anaesthetic) keeping all variables constant. Therefore pH data collected in a child within 24 hours of endoscopy under general anaesthesia are representative. PMID- 7492166 TI - Trichothiodystrophy with sideroblastic anaemia and developmental delay. AB - A patient with sideroblastic anaemia, development delay, and trichothiodystrophy is presented. Trichothiodystrophy is a feature of several autosomal recessive diseases. Photosensitivity, failure to thrive, and developmental delay are commonly observed in affected cases. X linked inheritance accounts for the bulk of cases with sideroblastic anaemia. This case highlights the importance of routine hair microscopy in cases of atypical ectodermal dysplasia. PMID- 7492165 TI - Skin collagen defects in a patient with juvenile hyaline fibromatosis. AB - Juvenile hyaline fibromatosis is a rare disorder characterised by multiple subcutaneous tumours, gum hypertrophy, muscle weakness, and flexion contractures of the large joints. Histology shows an abundance of a homogenous, amorphous, acidophilic extracellular matrix in which spindle shaped cells are embedded forming minute streaks. It has been previously suggested that collagen abnormalities may be involved. A 14 month old girl with this syndrome is described in whom postmortem western blot studies were performed. These studies revealed an absent pro-alpha 2(I) chain and an absent collagen type III chain in skin but not in the other organs examined. PMID- 7492167 TI - Growth hormone deficiency and empty sella in DIDMOAD syndrome: an endocrine study. AB - Two girls with DIDMOAD syndrome are presented. One also had severe megaloblastic sideroblastic anaemia and the other several neurological manifestations. Both were short with defective growth hormone secretion. Computed tomography revealed empty sella in both girls; one had widespread atrophic cortical and cerebellar changes. High doses of thiamine improved the anaemia in the first case, increased C peptide secretion in both, but had no effect on the neurological abnormalities. PMID- 7492169 TI - Osteopenia in Crohn's disease. AB - A 16 year old boy presented soon after diagnosis of Crohn's disease with diffuse osteopenia of the lumbar spine with collapse of three vertebral bodies and grossly reduced bone mineral density noted on dual energy x ray absorptiometry scan. Treatment of the underlying Crohn's disease and induction of puberty resulted in marked improvement in bone mineralisation. PMID- 7492168 TI - Management of infantile polyposis syndrome. AB - A boy with generalised infantile polyposis syndrome is reported to highlight the difficulties in management. Despite attempts to reduce polyp mass by regular endoscopic polypectomy, daily transfusions of blood products, and a trial of the non-steroidal anti-inflammatory agent sulindac, his condition gradually deteriorated and he died of septicaemia. PMID- 7492170 TI - Dopa responsive dystonia. AB - There may be insufficient awareness of dopa responsive dystonia (DRD), which has a characteristic diurnal variation of symptoms. Two children are reported in whom the diagnosis of DRD was missed. The first was thought to have hysteria and the second hereditary spastic paraparesis. A full history is vital for the diagnosis of this important treatable syndrome. PMID- 7492171 TI - Co-trimoxazole induced aseptic meningitis. AB - A 15 year old boy presented with two episodes of aseptic meningitis-like reactions after ingestion of co-trimoxazole. The diagnosis of co-trimoxazole induced aseptic meningitis was made. This syndrome should be considered in the differential diagnosis of aseptic meningitis. PMID- 7492172 TI - Cardiac involvement in mucopolysaccharidoses: effects of allogeneic bone marrow transplantation. AB - Echocardiography was performed in 16 children undergoing bone marrow transplantation (BMT) for mucopolysaccharidoses. Cardiac involvement before BMT was detected in seven (44%). One year after BMT (11 patients/five with cardiac involvement), left ventricular restriction resolved in 2/3 patients and hypertrophy in one. In the remainder, at mean follow up of 2.5 years, no progression of preexisting or development of new cardiac involvement was noted. It is concluded that in a significant proportion of patients with mucopolysaccharidoses, cardiac involvement improved after BMT. PMID- 7492174 TI - Core knowledge, skills, and attitudes in child health for undergraduates. PMID- 7492173 TI - Child health education for the year 2000. PMID- 7492176 TI - Core knowledge, skills, and attitudes for undergraduates: kindling curiosity. PMID- 7492175 TI - Teaching undergraduates child health: what, how, where, and when? PMID- 7492177 TI - Statistics from the inside. 16. Multiple regression (2). PMID- 7492179 TI - Evidence for superantigen mediated process in Kawasaki disease. PMID- 7492178 TI - Vitamin A supplementation in developing countries. PMID- 7492180 TI - Birth weight in phenylketonuria. PMID- 7492181 TI - Burkholderia cepacia and delta F508 homozygosity in cystic fibrosis. PMID- 7492182 TI - Diagnosis of mycobacterial lymphadenopathy. PMID- 7492183 TI - Compliance with growth hormone treatment--are they getting it? PMID- 7492184 TI - Morbidity from excessive intake of high energy fluids: the 'squash drinking syndrome'. PMID- 7492185 TI - Overnight oscillations of rectal temperature. PMID- 7492186 TI - The art of communication with children. PMID- 7492187 TI - Cardiac effects of growth hormone in short normal children. PMID- 7492188 TI - Acute respiratory infection: a global challenge. PMID- 7492189 TI - Otoacoustic emissions. PMID- 7492191 TI - Blood pressure and smoking: observations on a national cohort. AB - The reasons why adult smokers have lower blood pressure than non-smokers have not been determined. It is possible that low blood pressure might precede the onset of smoking. This study investigates this hypothesis in a national cohort study in Britain. Blood pressures and pulse rates taken on a sample of 5019 members of the British Birth Cohort Study (BCS 70) at the age of 10 years were analysed in relation to self reported smoking behaviour at age 16+ years. Prospectively, those children who had lower diastolic blood pressure or pulse rate at age 10 were more likely to have smoked by age 16+ years. Using analysis of variance, pulse rate was significantly related to smoking in young men (p < 0.001). Seventy per cent of those with lower pulse (below the 10th centile), 58% with medium pulse, and 52% with the higher pulse (above the 90th centile) had ever smoked by age 16+ years. In young women, pulse rate (p = 0.003), diastolic pressure (p = 0.024), and systolic pressure (p = 0.032) at age 10 were all significantly related to smoking at age 16. This longitudinal study found that lower blood pressure and slower pulse rate were related to the onset of smoking in children. More research is needed on this new observation. PMID- 7492190 TI - Size at birth and plasma insulin-like growth factor-1 concentrations. AB - OBJECTIVE: To test the hypothesis that reduced fetal growth leads to altered plasma insulin-like growth factor-1 (IGF-1) concentrations in childhood. DESIGN: A follow up study of 4 year old children whose birth weights were recorded, and of 7 year old children whose weight, length, head circumference, and placental weight were measured at birth. SETTING: Pune, India, and Salisbury, England. SUBJECTS: 200 children born during October 1987 to April 1989 in the King Edward Memorial Hospital, Pune, weighing over 2.0 kg at birth and not requiring special care, and 244 children born during July 1984 to February 1985 in the Salisbury Health District and still living there. MAIN OUTCOME MEASURE: Plasma IGF-1 concentrations. RESULTS: In both groups of children, and consistent with findings in other studies, plasma IGF-1 concentrations were higher in taller and heavier children, and higher in girls than boys. Allowing for sex and current size, concentrations were inversely related to birth weight (Pune p = 0.002; Salisbury p = 0.003). Thus at any level of weight or height, children of lower birth weight had higher IGF-1 concentrations. The highest concentrations were in children who were below average birth weight and above average weight or height when studied. Systolic blood pressures were higher in children with higher IGF-1 concentrations (Pune p = 0.01; Salisbury p = 0.04). CONCLUSIONS: Children of lower birth weight develop higher circulating concentrations of IGF-1 than expected for their height and weight. This is consistent with the hypothesis that under-nutrition in utero leads to reprogramming of the IGF-1 axis. The increase of plasma IGF-1 concentrations in low birthweight children may also be linked to postnatal catch up growth. High IGF-1 concentrations may be one of the mechanisms linking reduced fetal growth and high blood pressure in later life. PMID- 7492192 TI - Asthma, enuresis, and chronic illness: long term impact on height. AB - OBJECTIVE: To determine whether common conditions in early childhood, such as asthma and psychosocial illness (mainly enuresis), affect height during later childhood and in adult life. DESIGN: Longitudinal follow up of subjects in the 1958 British Birth Cohort Study. Data from the birth survey and ages 7, 11, 16, and 23 were used. SUBJECTS: 12,537 subjects remaining in the study at age 23, representing 76% of the target population, cohort members still alive and resident in Britain. RESULTS: Heights of children with allergic, acute or psychosomatic illness, or asthma/wheezy bronchitis did not differ by age 7 from those of children without such illnesses. When asthma was graded by severity, there was a trend (not significant) for the severe group to be shorter at ages 16 and 23. Although children with a chronic illness by age 7 were on average almost 0.5 cm shorter than children without such illnesses, this difference was reduced by half and was not significant after adjusting for maternal height, birth weight, parity, and social class at birth. However, a marked and long lasting effect was found for children with psychosocial illness who at age 7 were significantly shorter, by a mean of 0.77 cm. Within this group, enuretic children with a problem at age 11 were more than 1 cm shorter in adulthood, even allowing for other height related factors. CONCLUSIONS: Common childhood illnesses do not appear to affect height, either in the short or in the long term, although exceptions include chronic illness and enuresis. The value of height as an indicator of child health status in an industrialised country such as Britain requires further reassessment. PMID- 7492193 TI - Determination of body composition from skinfold thickness: a validation study. AB - Measurement of body composition is proving increasingly important in clinical nutrition and research. Skinfold thickness is a simple means of estimating body composition which is widely used in children, but there is little information on its validity. There has been a proliferation of equations for estimation of body composition from skinfolds, but some doubt as to their general applicability. The aim of the present study was to validate five currently used equations for this purpose in a sample of 98 healthy prepubertal children (64 boys, 34 girls), mean (SD) age 9.1 (1.7) years by comparison of estimates from each equation with measurements of fatness derived from hydrodensitometry. Differences between methods were determined by calculation of biases and limits of agreement. Limits of agreement between predicted and measured fatness were wide, particularly in the girls, and some distinct biases were apparent. Choice of prediction equation therefore has a substantial influence on the estimate of fatness obtained when using skinfolds in children. The existing published equations are associated with large random errors or significant systematic errors. For the time being skinfolds might best be regarded as indices (rather than measures) of body fatness in individuals, or means of estimating body fatness of groups. Estimating the total body fatness of individual prepubertal children using skinfolds, on the basis of this evidence, is not advisable at present. PMID- 7492194 TI - Long term sequelae of sex steroid treatment in the management of constitutionally tall stature. AB - AIM: To evaluate possible long term side effects of high doses of sex steroids in the management of constitutionally tall stature, with special attention to hypothalamic-gonadal function. METHODS: Sixty four tall adult men and 180 tall adult women, who received supraphysiological doses of sex hormones during puberty, were interviewed in a standardised way at a mean follow up period of 10 years after cessation of treatment. Sixty one untreated tall adult men and 94 untreated tall adult women served as controls. RESULTS: The majority of the subjects were satisfied with their decision regarding hormone treatment. Seventy seven per cent of the women and 78% of the men reported one or more side effects during treatment. Most side effects were mild. In women, only 3% stopped treatment because of an adverse event; in men, the reported side effects never stopped treatment. The frequency of reported side effects in women was higher during treatment with high doses of oestrogens than during oral contraceptive use, indicating a dose dependent relationship. Amenorrhoea of longer than six months after cessation of therapy was found in 5%. Menstrual cycle characteristics of previously treated women were comparable with controls. Malignancy was not reported. Information about a total of 127 pregnancies was obtained and revealed no distinct differences in details and outcome between previously treated women and men, and controls. CONCLUSIONS: At a mean follow up of 10 years there is no evidence that pharmacological doses of sex hormones have a long term effect on reproductive function. However, this period is still too short to draw definite conclusions. PMID- 7492195 TI - Factors associated with lowered intelligence in homozygous sickle cell disease. AB - The intelligence quotient (IQ) of 60 patients with homozygous sickle cell (SS) disease and 60 age and sex matched controls with a normal haemoglobin (AA) genotype aged 15-18 years, followed up in a cohort study from birth, was assessed by the Wechsler intelligence scales for children or for adults. IQ appeared to be normally distributed in both genotypes but mean values in SS disease were 5.6 points (95% confidence interval (CI) 1.0 to 10.2) lower than in AA controls (p = 0.016). The difference occurred in both verbal (5.5 points, p = 0.017) and performance (5.0 points, p = 0.044) subscales of the IQ score and the IQ defect in SS disease was associated with a significantly lower attention factor score (p = 0.005) but not with other factor scores. The genotype difference in IQ was not accounted for by differences in parental occupational level, school absenteeism, or school drop out, or reported activity level. In SS disease, IQ was not related to mean steady state haemoglobin, fetal haemoglobin, or mean cell haemoglobin concentration, or clinical severity as judged by the frequency of painful crises, hospital admission, or sick visits. IQ, at age 15-18 years, correlated with the patients' height at all ages from 1 to 10 years (partial correlations increasing from 0.14 (p = 0.15) at age 1 to 0.27 (p = 0.004) at age 10). Adjusting for height reduced the mean genotype difference in IQ to 5.5 (95% CI 0.6 to 10.3) points at age 1 and to 2.6 points (95% CI to -2.3, 7.5) at age 10. Prepubertal height therefore accounted for much of the genotype difference in IQ. It is speculated that early factors, possible nutritional, contribute to both impaired growth and mental development in sickle cell disease. PMID- 7492196 TI - Asthma knowledge, attitudes, and quality of life in adolescents. AB - Adolescents with asthma, their peers, and their teachers were studied in order to establish the level of knowledge concerning asthma and its management, their attitudes towards asthma, and the degree quality of life impairment due to asthma. A community survey was conducted among year 8 high school students (n = 4161) and their teachers (n = 1104). There was a good response rate to the questionnaires from students (93%) and teachers (61%). Twenty three per cent of students had asthma and this caused mild to moderate quality of life impairment, particularly with strenuous exercise. Asthma was provoked by passive smoke exposure in 30% of asthmatic students and up to 51% of students avoided situations because of asthma triggers. Asthma knowledge was low in teachers (mean score 14.90 out of a possible 31), students without asthma (11.25) and students with asthma (14.50). Specific knowledge on the prevention and treatment of exercise induced asthma was poor. There was a moderate degree of tolerance towards asthma among all three groups. Most considered internal locus of control as important, although students without asthma also considered chance to be a determinant of outcomes for people with asthma. Asthma is a common cause of quality of life impairment among year 8 high school students. Although specific knowledge on asthma is low, students and teachers hold favourable attitudes towards asthma. There are opportunities to intervene and improve asthma management among adolescents. PMID- 7492200 TI - Problems of differentiation between epilepsy and non-epileptic paroxysmal events in the first year of life. AB - Twenty two babies under 1 year old were referred for evaluation of suspected epileptic seizures. Nine were found to have epilepsy. In the other 13--all developing normally, aged up to 10 months--the spells were non-epileptic paroxysmal events (NEPE). They consisted of five patterns of movement: (1) eye blinking; (2) 'no' movements; (3) body posturing with head and arm jerks; (4) masturbation-like movements; and (5) myoclonic head flexion. The NEPE were present for a period of two weeks to seven months. Although some NEPE cannot be clinically differentiated from true epilepsy, in these infants at least four interictal EEGs were normal, the spells completely resolved after a relatively short period without antiepileptic treatment, and the infants continued to develop normally with no evidence of epilepsy during a follow up period of 28 to 38 months. This sample indicates that the frequency of NEPE in the first year of life may be high. Cautious clinical consideration, repeat EEGs and, when appropriate, a few weeks' observation are recommended. Awareness of these benign behavioural spells in this young age group is important, and parents can be reassured. Nevertheless, the spells may illustrate a 'foggy frontier' between NEPE and epilepsy. The lack of evidence for any other disease process in affected infants, as well as the disappearance of the NEPE without any intervention, indicates that a maturational process may be involved. PMID- 7492197 TI - The effects of inhaled beclomethasone dipropionate on lung function and histamine responsiveness in recurrently wheezy infants. AB - Inhaled steroids improve pulmonary function and bronchial responsiveness in older asthmatics. Data from studies using subjective outcome measures to determine the effectiveness of inhaled steroids in infants with recurrent wheezing are equivocal. Therefore, this study tested the hypothesis that beclomethasone dipropionate improves pulmonary function, including bronchial responsiveness to histamine, in recurrently wheezy infants. The study was double blind, placebo controlled lasting nine weeks. After the first baseline week, pulmonary function was measured using the rapid thoracoabdominal compression technique and bronchial responsiveness assessed with a histamine challenge test. Infants were then randomly allocated to receive doses of placebo or beclomethasone dipropionate (100 micrograms/puff) from metered aerosols. Two puffs of test aerosol were administered twice daily for eight weeks via a large volume spacer fitted with a facemask. Symptoms were recorded daily and pulmonary function and bronchial responsiveness assessed at the end of the treatment period; 50 infants, median age 12 months (range 5 to 18 months), were recruited. Twenty three in the beclomethasone dipropionate group and 15 in the placebo group completed the study and had pairs of pulmonary function measurements. Three were probable treatment failures (one beclomethasone dipropionate, two placebo), three were possible treatment failures (placebo), and others were non-compliant with study protocol. Baseline variables were not significantly different between those infants who completed the study and those who did not. Beclomethasone dipropionate and placebo groups were similar in all respects at baseline. Lung function and symptoms improved for both groups of infants during the study. Bronchial responsiveness increased significantly in the placebo group but there were not statistically significant differences between groups for any of the other outcome measures. It is concluded that beclomethasone dipropionate (400 microgram daily) via a large volume spacer does not significantly improve lung function or symptoms in recurrently wheezy infants but might hav a beneficial effect on bronchial responsiveness. PMID- 7492199 TI - Cerebrospinal fluid zinc concentrations in febrile convulsions. AB - Zinc modulates the activity of glutamic acid decarboxylase, the rate limiting enzyme in the synthesis of gamma-aminobutyric acid (GABA), which is a major inhibitory neurotransmitter. Low cerebrospinal fluid GABA values have been reported in association with several seizure disorders, including febrile convulsions. It is also known that fever and/or infections may cause a reduction in serum zinc concentrations. In this study the hypothesis that febrile convulsions are related to low cerebrospinal fluid zinc was tested. Cerebrospinal fluid zinc concentrations were measured in 66 febrile children: 32 with febrile convulsions, 18 with fever but without convulsions, and 16 with aseptic (viral) meningitis. There was no statistically significant difference in the cerebrospinal fluid zinc between the three groups of children, and the mean concentration was 26.2 micrograms/l. No significant relationship was found between either age, gender, maximal temperature, type of infection, or time of performance of the lumbar puncture and cerebrospinal fluid zinc concentration. These results do not support the hypothesis that febrile convulsions are related to reduced cerebrospinal fluid zinc concentrations. PMID- 7492198 TI - Blood eosinophil counts and arterial oxygen tension in acute asthma. AB - OBJECTIVE: To investigate whether during acute asthma episodes a decrease in blood eosinophil count could correlate with the severity of the disease. DESIGN: Prospective study on paediatric asthmatic patients admitted for acute asthma exacerbation between January 1992 and August 1993. All patients were regularly followed up in an outpatient clinic and had had a complete clinical evaluation < 1 month before admission. SETTING: Pulmonary division of the G Gaslini paediatric research institute, Genoa, Italy. SUBJECTS: 21 asthmatic patients, 59 (SEM 9) months of age, admitted for acute asthma exacerbation. On the basis of clinical evaluation and the results of blood and microbiological tests performed during acute asthma exacerbations, patients were divided into two subgroups: infected (n = 13) and non-infected (n = 8). RESULTS: All but one of the patients showed a marked decrease in blood eosinophil count during the acute asthma episode, in comparison with recent count (< 1 month before admission) obtained in clinically stable conditions: 662 (116) v 210 (54) eosinophils/mm3, p < 0.0003. The decrease in the eosinophil count was more pronounced in the infected patients than in the non-infected patients, but the difference was not statistically significant (p > 0.05). Similarly, transcutaneous arterial oxygen pressure (PaO2) values measured during acute asthma exacerbations tended to be lower in infected patients, without, however, reaching statistical significance: 8.6 (0.7) v 10.1 (0.9) kPa, p > 0.05). The correlation between the decrease in blood eosinophil count and PaO2 during the acute asthma exacerbations was significant in all the patients (r2 = 0.235, p = 0.022) and in the non-infected patients (r2 = 0.653, p = 0.015), but not in infected patients. In this latter subgroup, a significant negative correlation was found between blood neutrophil counts during acute asthma exacerbations and PaO2 (r2 = 349, p = 0.026). CONCLUSIONS: During acute asthma exacerbations in atopic patients without clinical evidence of infection, the decrease in blood eosinophil count correlates significantly with the decrease in PaO2, further supporting the role of eosinophils in allergic asthma. PMID- 7492201 TI - Intracranial tumours during the first two years of life: presenting features. AB - Between 1979 and 1994, 21 children (nine females, 12 males) with intracranial tumours diagnosed before the age of 2 years (range 2-23 months) were treated at the University Hospital of Wales. The commonest presenting symptoms were vomiting (n = 9) and unsteadiness (n = 8); the commonest presenting sign was enlarged occipitofrontal circumference (> 97th centile in 16 and > 90th centile in a further two). In five cases with signs of raised intracranial pressure, meningitis was the clinical diagnosis, and a lumbar puncture was performed. For cases with long delays in diagnosis, multiple other disorders had been considered and the significance of head enlargement had not been recognised. In very early childhood, intracranial tumours are uncommon and can mimic other disorders, especially meningitis. Early neuroimaging is advised when a child presents with recent onset of neurological symptoms and a disproportionately large head. PMID- 7492202 TI - Nutritional care and candidates for small bowel transplantation. AB - Twenty two children were evaluated for small bowel transplantation between 1989 and 1994. Eight were unfit for transplantation and died within three months; a raised plasma bilirubin concentration (> 200 mumol/l) predicted death from liver failure within six months (p = 0.0001). The 11 children who were not managed by a multidisciplinary nutritional care team were less well nourished at referral, had more complications with intravenous feeding catheters, and greater early mortality while awaiting transplantation (p < 0.05). It is recommended that children with chronic intestinal failure be referred for assessment early, before liver dysfunction is established. PMID- 7492204 TI - Indications for investigation of chronic gastrointestinal symptoms. AB - Simple routine blood tests (full blood count, platelet count, erythrocyte sedimentation rate, C reactive protein, and serum albumin) may select children with chronic gastrointestinal symptoms who require endoscopic assessment and are thus an important aid in the prediction of endoscopic status. When all of the results are normal then chronic inflammatory bowel disease is an unlikely diagnosis. PMID- 7492203 TI - Filter paper cortisol profiles in secondary adrenocortical insufficiency. AB - The use of filter paper four point 24 hour cortisol profiles in the diagnosis and management of secondary adrenocortical deficiency was examined. Eighteen children with familial short stature and isolated growth hormone deficiency had normal 24 hour and morning plasma cortisol concentrations measured. Eight of 11 children with multiple pituitary hormone deficiencies had evidence of hypocorticalism despite previously normal baseline cortisols or responses to insulin hypoglycaemia or tetracosactrin. Nine of 11 children with hypopituitarism on replacement treatment (4.9-12.5 mg/m2/day) had satisfactory concentrations of cortisol, though morning cortisols were often low. Filter paper profiles are a simple, inexpensive, and relatively non-invasive way of managing children with suspected hypocorticalism and of monitoring their treatment. PMID- 7492205 TI - Family meals: a thing of the past. PMID- 7492206 TI - Adult learning theory, problem based learning, and paediatrics. PMID- 7492207 TI - Data collection and management for research studies. PMID- 7492208 TI - Maintenance treatment and shared care in lymphoblastic leukaemia. PMID- 7492209 TI - Survival data. PMID- 7492210 TI - Maternal smoking and blood pressure in 7.5 to 8 year old offspring. PMID- 7492211 TI - Plasma interleukin-3 and interleukin-4 concentrations in Turkish asthmatic children. PMID- 7492212 TI - A judicial comment on temporary brittle bone disease. PMID- 7492213 TI - Treatment response in Paget's disease. PMID- 7492214 TI - What you need to know when you go with the flow: pitfalls in the use of flow cytometry. PMID- 7492215 TI - Rheumatological manifestations of haematological diseases. PMID- 7492216 TI - Subcutaneous nodules in a patient with polyarthritis. PMID- 7492217 TI - Risk of vertebral fracture and relationship to bone mineral density in steroid treated rheumatoid arthritis. AB - OBJECTIVES: To determine the prevalence of vertebral fracture in postmenopausal women with steroid treated rheumatoid arthritis (RA), and whether the risk of vertebral fracture could be predicted from measurements of bone mineral density (BMD). METHODS: Vertebral deformities were defined from spine radiographs in 76 postmenopausal women with steroid treated RA (aged 50-79 years) and 347 age matched women from a population based group, using a morphometric technique. Lumbar spine (LS) BMD was measured by dual energy x ray absorptiometry. RESULTS: The odds ratio for vertebral fracture in the women with RA was 6.2 (95% confidence interval 3.2 to 12.3). The decrease in LS-BMD was less than expected for the observed prevalence of vertebral fracture and, among the women with RA, LS-BMD was not lower in those with vertebral fractures. CONCLUSIONS: We conclude that patients with steroid treated RA may have abnormal bone quality, and that LS BMD cannot be used to predict the risk of vertebral fracture in these patients. PMID- 7492218 TI - Cut off values of bone mineral density defining postmenopausal women with bone fractures. AB - OBJECTIVES: To define cut off values of bone mineral density (BMD), measured at different forearm sites by single photon absorptiometry, that discriminated postmenopausal women who have had fractures from those who have not, in order to provide an index of fracture risk for screening purposes. METHODS: BMD values were measured at distal third radius (DTR) and ultradistal radius (UDR) sites in two groups of postmenopausal women. Group 1 (n = 398) had not suffered fractures, and group 2 (n = 354) had appendicular fractures. Results were evaluated using receiver operating characteristic (ROC) curves. RESULTS: BMD values were significantly greater (p < 0.001) at both sites in group 1. Weight was lower in group 2 than in group 1 (p < 0.001). The BMD cut off value was 0.596 g/cm2 at DTR (sensitivity 63%, specificity 72%), and 0.310 g/cm2 at UDR (sensitivity 80%, specificity 80%). Values of area under ROC curves estimated for UDR were greater than for those DTR (p < 0.001). CONCLUSION: For screening purposes the BMD cut off value of 0.310 g/cm2 at the UDR may be useful in identifying women at high risk of fracture in an unselected postmenopausal population. BMD measured at UDR appeared to have a better predictive value than that at DTR. PMID- 7492219 TI - Rheumatoid factor, chronic arthritis and mortality. AB - OBJECTIVE: To investigate chronic arthritis and rheumatoid factor (RF) for their prediction of premature total and cardiovascular mortality. METHODS: In 1978-80, a representative population sample of 8000 Finns aged 30 or more was invited to participate in a comprehensive health examination; 90% complied. Arthritis was diagnosed on the basis of medical history, symptoms, and physical examination. Serum RF was determined by the sensitised sheep cell agglutination test. RESULTS: By the end of 1992 1597 of the subjects had died from all causes, including 876 deaths from cardiovascular diseases. When adjusted for age, gender and smoking, the relative risk of persons with RF positive arthritis dying from any cause was 1.61 (95% confidence interval (CI) 1.03 to 2.51); RF negative non-erosive arthritis was not associated with mortality (relative risk 1.03; 95% CI 0.72 to 1.49). In the absence of arthritis, 'false positive' RF titres > or = 128 predicted cardiovascular deaths with a relative risk of 1.74 (95% CI 1.06 to 2.86). CONCLUSION: Both RF positive arthritis and false positive RF reactions predict mortality, but through different disease patterns. PMID- 7492220 TI - Use of high resolution computed tomography of the lungs in patients with rheumatoid arthritis. AB - OBJECTIVE: To assess the usefulness of high resolution computed tomography (HRCT) of the lungs in patients with rheumatoid arthritis (RA) with and without respiratory symptoms. PATIENTS AND METHODS: Eighty eight RA patients with a mean duration of disease 12 (SD 8) years were evaluated. Eleven patients were excluded because of previous exposure to silica. The 77 remaining patients formed two groups according to the absence (group I, n = 38) or the presence (group II, n = 39) of chronic respiratory symptoms. A control group consisted of 51 non-smoking, healthy patients. RESULTS: The most frequent abnormalities observed in the 77 RA patients were bronchiectasis or bronchiolectasis (n = 23, 30%), pulmonary nodules (n = 17, 22%), subpleural micronodules or pseudoplaques (n = 13, 17%), ground glass opacities (n = 11, 14%), and honeycombing (n = 8, 10%). Bronchiectasis or bronchiolectasis (p = 0.012), rounded opacities (p = 0.016), ground glass attenuation (p = 0.004), and honeycombing (p = 0.002) were found more often in RA group II (with respiratory symptoms) than in group I (no respiratory symptoms). Non-linear septal opacities were more frequent in group I than in the control group, but other HRCT findings did not differ statistically significantly between group I and the control group. CONCLUSION: Bronchiectasis may be a characteristic lung change in RA patients. Abnormalities on HRCT are less frequently observed in the absence of respiratory symptoms than in the presence of such symptoms (29% versus 69%). PMID- 7492221 TI - Urinary nitrate excretion is increased in patients with rheumatoid arthritis and reduced by prednisolone. AB - OBJECTIVES: To determine daily production of nitric oxide (NO) measured as urinary nitrate excretion, and the effect of prednisolone in patients with rheumatoid arthritis (RA). METHODS: Twenty four hour urinary nitrate was measured by gas chromatography in 10 patients with RA, before and two to four weeks after commencement of prednisolone 0.5 mg/kg body weight, and in 18 healthy controls. RESULTS: Before the start of prednisolone treatment the urinary nitrate excretion in patients with RA was 2.7-fold greater (p < 0.001) than that in healthy volunteers. After prednisolone it decreased significantly, by 28%, at which time inflammatory activity (as indicated by C reactive protein, erythrocyte sedimentation rate, joint count, and early morning stiffness) was also reduced considerably. Despite this decrease, the urinary nitrate excretion in patients with RA remained twice that in the control group (p < 0.05). CONCLUSIONS: Our data suggest that the endogenous production of NO is enhanced in patients with RA. Furthermore, the results indicate that, in parallel with suppression of inflammation, this increased NO synthesis could be reduced by prednisolone treatment. PMID- 7492222 TI - Evidence for a causal relationship between the structure, size, and load of calcium pyrophosphate dihydrate crystals, and attacks of pseudogout. AB - OBJECTIVE: To investigate any relationship between the nature, size, and numbers of synovial fluid (SF) calcium pyrophosphate dihydrate (CPPD) crystals, and attacks of pseudogout. METHODS: Knee SF was aspirated from nine selected patients, first during an attack of pseudogout (acute sample) and again later when the attack had subsided (interval sample). CPPD crystals were extracted, weighed, examined by high resolution transmission electron microscopy (HRTEM), and characterised by size and crystal habit (monoclinic or triclinic). Structural analysis was carried out by x ray powder diffraction (XRD) and the proportions of monoclinic to triclinic CPPD were estimated from densitometric measurements of selected key reflections. RESULTS: The mean crystal size, by HRTEM, indicated that the crystals in the acute sample were larger than those in the interval sample. The ratio of monoclinic to triclinic CPPD, whether estimated from their morphological appearance by HRTEM, or from XRD, was greater in the acute than in the interval sample in all nine patients. The total amount of extracted mineral varied, but in every patient the concentration of CPPD per ml of fluid, and the total mineral per joint, were greater in the acute sample than in the interval sample. CONCLUSION: In this highly selected group of patients, the large numbers of CPPD crystals associated with attacks of pseudogout included a greater proportion of monoclinic crystals, and larger crystals, than those present when inflammation had subsided. A special, phlogistic population of crystals may exist, originating in different joint tissues, or cleared in a different manner, than the more common populations of smaller crystals with a greater proportion of triclinic CPPD, seen in chronic disease. PMID- 7492223 TI - Proteoglycan depletion and size reduction in lesions of early grade chondromalacia of the patella. AB - OBJECTIVE: To determine the content and molecular size of proteoglycans (PGs) in patellar chondromalacia (CM) and control cartilages as a first step in investigating the role of matrix alterations in the pathogenesis of this disease. METHODS: Chondromalacia tissue from 10 patients was removed with a surgical knife. Using identical techniques, apparently healthy cartilage of the same site was obtained from 10 age matched cadavers (mean age 31 years in both groups). Additional pathological cartilage was collected from 67 patients with grades II IV CM (classified according to Outerbridge) using a motorised shaver under arthroscopic control. The shaved cartilage chips were collected with a dense net from the irrigation fluid of the shaver. The content of tissue PGs was determined by Safranin O precipitation or uronic acid content, and the molecular size by mobility on agarose gel electrophoresis. RESULTS: The mean PG content of the CM tissue samples with a knife was dramatically reduced, being only 15% of that in controls. The cartilage chips collected from shaving operations of grades II, III, and IV CM showed a decreasing PG content: 9%, 5%, and 1% of controls, respectively. Electrophoretic analysis of PGs extracted with guanidium chloride from the shaved tissue samples suggested a significantly reduced size of aggrecans in the mild (grade II) lesions. CONCLUSION: These data show that there is already a dramatic and progressive depletion of PGs in CM grade II lesions. This explains the softening of cartilage, a typical finding in the arthroscopic examination of CM. The PG size reduction observed in grade II implicates proteolytic attack as a factor in the pathogenesis of CM. PMID- 7492224 TI - Proinflammatory cytokine production and cartilage damage due to rheumatoid synovial T helper-1 activation is inhibited by interleukin-4. AB - OBJECTIVES: To investigate the role of T helper-1 cell (Th1) activation in the induction of proinflammatory cytokine production and cartilage damage by rheumatoid arthritis (RA) synovial fluid mononuclear cells (SFMNC) and the subsequent possible beneficial role of the T helper-2 cell (Th2) cytokine interleukin-4 (IL-4) in the inhibition of this process. METHODS: SFMNC were stimulated with bacterial antigen (hsp60) to activate Th1 cells. Th1 and Th2 specific cytokine profiles (interferon gamma (IFN gamma) and IL-4) and proinflammatory cytokines interleukin-1 (IL-1) and tumour necrosis factor alpha (TNF alpha) in the conditioned media were analysed. In addition, the conditioned media were tested for their ability to induce cartilage damage. The same parameters were measured in the presence of IL-4. RESULTS: Stimulation of SFMNC with bacterial antigen resulted in an increase in IFN gamma, IL-1, and TNF alpha production which was accompanied by the induction of cartilage damage. Th1 activation could be inhibited by IL-4 as shown by a reduction of IFN gamma. This was accompanied by a decrease in IL-1 and TNF alpha production and inhibition of cartilage damage. CONCLUSIONS: Th1 activation is a possible mechanism by which inflammation in RA joints is enhanced. The Th2 cytokine IL-4 inhibits this Th1 activity and may diminish inflammation and induction of cartilage damage in RA joints. PMID- 7492225 TI - Differential distribution of annexins-I, -II, -IV, and -VI in synovium. AB - OBJECTIVES: To examine the distribution of four annexins in non-inflamed rheumatoid arthritic and osteoarthritic synovial tissue. METHODS: Frozen sections were stained with monoclonal antibodies (MAb) specific for annexins-I, -II, -IV, and -VI, and for cell lineage related markers including CD68 and CD14 (macrophages), prolyl hydroxylase (fibroblasts), and CD3 (T cells). RESULTS: Each of the annexins was present in synovial tissues in significant amounts in the three groups studied. Annexin-I was predominantly found within the synovial lining layer and double labelling showed it to be present predominantly in cells of the macrophage lineage. In rheumatoid specimens there was increased staining within the lining layer, perivascularly and on macrophages within the tissue stroma. Annexin-II was present in a distribution similar to that of annexin-I, but with more prominent perivascular staining. Annexins-IV and -VI were seen chiefly in association with areas of lymphocyte infiltration in rheumatoid tissue, whereas annexins-I and -II were absent from these areas. Endothelial cells stained weakly positive for annexins-I and -II, and more strongly for -IV and -VI. CONCLUSIONS: This study demonstrates that annexins (particularly annexin I, a putative mediator of the anti-inflammatory activities of glucocorticoids) are abundant in rheumatoid and non-rheumatoid synovial tissue, annexins-IV and VI having a distribution distinct from that of -I and -II. PMID- 7492226 TI - Treatment of refractory juvenile chronic arthritis by monoclonal CD4 antibodies: a pilot study in two children. AB - OBJECTIVE--To study the effect of anti-CD4 treatment in patients suffering from refractory systemic onset juvenile chronic arthritis (JCA). METHODS--Two children were treated with the mouse CD4 antibody MAX.16H5. The effects on numbers of circulating CD4 T cells, clinical symptoms and C reactive protein (CRP) level were studied and the appearance of human antimouse immunoglobulin antibodies investigated. RESULTS--In patient one, disappearance of fever and malaise and a reduction of arthritic activity were observed together with a reduction in CRP. When disease activity returned eight weeks later, a second successful course of treatment was administered. It was possible to reduce the corticosteroid dosage permanently. In the other child, a first treatment cycle did not alter disease activity. A marked reduction in clinical and laboratory disease activity markers was observed after the second course. Only transient and mild side effects were observed. One patient exhibited a short lasting febrile reaction with chills, the other an urticarial rash. In both patients, human antibodies to mouse immunoglobulin became detectable. The decrease in the number of CD4 T cells in the peripheral blood was only short lasting and numbers returned to normal values within one to eight weeks, even after the second course of antibody treatment and under concomitant immunosuppressive treatment. No sustained clinical remissions could be achieved. CONCLUSIONS--These preliminary observations support the evidence of positive effects of CD4 antibody treatment in refractory systemic onset JCA. Long term efficacy, however, remains to be established. PMID- 7492227 TI - Comparison of clinical and self reported diagnosis for rheumatology outpatients. AB - OBJECTIVE: To examine the sensitivity of patient self reported diagnoses compared with physician diagnoses in a rheumatology outpatient population. METHODS: A mailed survey to 472 rheumatology outpatients (81% response rate) asked about joint symptoms, disabilities, and underlying rheumatic conditions. The self reported diagnoses were linked with physician diagnoses in the rheumatology clinic computer based diagnostic registry. RESULT: Overall there was an 87% sensitivity for self reported compared with physician diagnoses when the matching criteria included compatible yet different diagnoses such as rheumatoid arthritis (RA) and osteoarthritis (OA). The sensitivity for exact match was 65%, and it varied with the underlying clinical diagnosis, and was greatest for RA (90%) and ankylosing spondylitis (AS) (100%), and intermediate for OA (52%) and psoriatic arthritis (50%). The sensitivity of self report was primarily related to the type of diagnosis (RA or AS v other rheumatic conditions; odds ratio = 16.3, 95% confidence interval (CI) 9.0 to 29.5), and also to difficulty in activities of daily living (odds ratio = 2.3, 95% CI 1.1 to 4.6) but not age, gender, duration of disease, or clinic attendance, as shown by multivariate analysis. CONCLUSIONS: This study in a rheumatology outpatient population indicated that most patients report a diagnosis which is compatible with the clinical diagnosis. These findings give an upper limit to the sensitivity of self reported diagnoses, though further research is needed to assess the extent to which our results may be generalised to other settings. PMID- 7492228 TI - Increased prevalence of symptomatic macrovascular disease in systemic sclerosis. AB - OBJECTIVES: To determine the prevalence of symptomatic macrovascular disease, as defined by the World Health Organisation questionnaire for intermittent claudication, in patients with systemic sclerosis (SSc), and to compare the prevalence in this population with a cross sectional population study group reported in the Edinburgh Artery Study (EAS). METHODS: A group of 53 patients with a diagnosis of limited or diffuse systemic sclerosis were identified from our register. They were asked to complete the WHO questionnaire to establish the presence or absence of intermittent claudication. In addition, each patient's case notes were reviewed to establish the existence of definite peripheral vascular disease and to determine their risk factor profile. RESULTS: Forty six patients responded to the questionnaire, giving a response rate of 87%. Intermittent claudication was found in 10 SSc patients (21.7%) according to the questionnaire (two with diffuse and eight with limited SSc), compared with a prevalence rate of 4.6% for claudication in the EAS. Three SSc patients experienced clinical events attributable to occlusion of a major artery proven on angiography. Four patients had hypertension, there were three current and four ex smokers, and two had increased total cholesterol. None of these patients had diabetes. CONCLUSIONS: This study demonstrated a greater prevalence of macrovascular disease in patients with SSc than had been found in a neighbouring population. PMID- 7492230 TI - Different concepts of musculoskeletal pain. PMID- 7492229 TI - Myocardial gallium-67 citrate scintigraphy in patients with systemic sclerosis. AB - OBJECTIVE: To detect myocardial involvement in 16 patients with systemic sclerosis using radiogallium scintigraphy METHODS: Sixteen patients with systemic sclerosis were investigated by myocardial gallium-67 citrate scintigraphy. Echocardiographic studies and (in the positive cases) magnetic resonance imaging were also performed. RESULTS: Increased myocardial gallium uptake was found in five of the 16 patients. None of these positive cases exhibited any 'classical' clinical, electrocardiographic or laboratory signs characteristic of myocarditis. The mean age of the positive cases was 60.8 (SD 5.0) years compared with 44.6 (10.7) years for the negative cases; no difference was detected in the mean disease duration between these two groups. Two of the affected cases belonged to the limited scleroderma subgroup. CONCLUSIONS: In spite of the low number of patients investigated, it is tempting to speculate that elderly patients with a disease onset around 60 years tend to be the candidates in whom a positive gallium-67 scintigraphy test may best indicate a silent myocardial interstitial inflammation. PMID- 7492231 TI - Activated protein C resistance caused by factor V Arg 506-->Gln mutation has no role in thrombotic manifestations of Behcet's disease. PMID- 7492232 TI - Exercise in primary prevention of osteoporosis in women. PMID- 7492233 TI - Economic evaluation in rheumatology: a necessity for clinical studies. PMID- 7492234 TI - Bone mineral density and bone turnover in spinal osteoarthrosis. AB - OBJECTIVES: To determine whether there was a generalised increase in bone mineral density (BMD) in spinal osteoarthrosis (OA), and to determine the mechanism of this possible protection against osteoporosis as assessed by biochemical markers of bone turnover. METHODS: We studied 375 women (ages 50 to 85) from a population based group. Spinal OA was defined from radiographs as the presence of degenerative changes affecting intervertebral or facet joints. BMD of the lumbar spine (LS), femoral neck (FN) and total body (TB) was measured by dual energy x ray absorptiometry (Lunar DPX). Bone turnover rates were estimated from measurement of biochemical markers of bone formation and resorption (urine deoxypyridinoline (Dpyr) and serum bone specific alkaline phosphatase (BAP)). RESULTS: BMD at each site was greater in the women with spinal OA (mean increase in LS-BMD 7.9%, 95% confidence interval (CI) 1.0 to 15.1; TB-BMD 8.4%, 95% CI 1.9 to 9.7; FN-BMD 6.4%, 95% CI 0.3 to 12.6). Twenty four hour urinary excretion of Dpyr, corrected for TB bone mineral content, and serum BAP were 19% lower in the women with spinal OA (95% CI for Dpyr 4.3 to 31.9%; for BAP 6.3 to 32.0%). CONCLUSIONS: Spinal OA is associated with a generalised increase in BMD and a decreased rate of bone turnover. This suggests that the protective effect of spinal OA against osteoporosis may be mediated by decreased bone turnover. PMID- 7492235 TI - Accuracy and precision of joint space width measurements in standard and macroradiographs of osteoarthritic knees. AB - OBJECTIVES: To improve the reproducibility and accuracy of joint space width (JSW) measurement as an assessment of cartilage loss in patients with osteoarthritis (OA) of the knee by determining how precision and accuracy of JSW measurement were altered by a computerised method of measurement, correction for radiographic magnification, radiography of the knee in the standing semiflexed view, and high definition macroradiography of the knee in the semiflexed view- taking JSW measurements from standard radiographs of OA knees in the extended view as the standard for comparison. METHODS: Twenty five OA and 10 nonarthritic knees were radiographed in the extended view and minimum JSW was measured manually. Conventional and x5 macroradiographs were taken in the semiflexed view. All radiographs were taken twice on the same day and repeated two weeks later. Automated computerised measurement of minimum JSW was obtained from digitally stored images of all radiographs. RESULTS: For medial compartment JSW measurements, computerised was more accurate than manual, correction for radiographic magnification improved precision and accuracy, measurements in the semiflexed view were more precise and accurate, and macroradiography increased measurement precision. For the lateral compartment JSW measurements, correction for radiographic magnification improved precision and accuracy, and the semiflexed view improved precision only. CONCLUSIONS: Protocols defining radiographic and mensural procedures are essential for quality control of knee radiography in the semiflexed view to permit accurate and reproducible measurement of JSW. Macroradiography provides greater precision of JSW measurement. PMID- 7492236 TI - Survival analysis of disease modifying antirheumatic drugs in Spanish rheumatoid arthritis patients. AB - OBJECTIVES: To evaluate the duration of treatment and the reasons for discontinuing therapy with disease modifying antirheumatic drugs in Spanish rheumatoid arthritis patients. METHODS: An observational study was made of 629 patients with rheumatoid arthritis treated with disease modifying antirheumatic drugs between 1979 and 1991. The outcomes (treatment termination because of toxicity and lack of response) of 991 treatment starts with intramuscular gold salts, D-penicillamine, azathioprine, and methotrexate were subjected to survival analysis. Cumulative probability of continuation of each drug (drug survival) was calculated by the Kaplan-Meier method and comparison between the survival curve of each was made by log rank testing. RESULTS: Median drug survival (95% confidence interval) was 51 (25-76.9) months for methotrexate, 39.9 (19.9-48.2) months for azathioprine, 34.9 (29.4-41.4) months for gold salts, and 16.4 (13.9 21) months for D-penicillamine. The highest cumulative probability of drug survival at five years was for methotrexate (45%); that at 10 years was for gold salts (15%). Up to 60% of the patients discontinued D-penicillamine in the first two years. Lack of response was the major limiting factor for all drugs except D penicillamine, for which it was toxicity. D-Penicillamine was associated with a greater rate of discontinuations because of toxicity in women and patients older than 65. Previous disease modifying antirheumatic drug administration did not influence current drug survival. CONCLUSION: Overall, gold salts remain useful for the treatment of rheumatoid arthritis over long periods of time in the population studied. Because of the high rate of continuation of treatment (survival) and the optimal efficacy and toxicity profiles observed with methotrexate after five years of treatment, it should be the drug of first choice for second line treatment of these RA patients. PMID- 7492237 TI - Application of markers of collagen metabolism in serum and synovial fluid for assessment of disease process in patients with rheumatoid arthritis. AB - OBJECTIVE: To assess the potential of markers of collagen metabolism to reflect disease processes in rheumatoid arthritis (RA). METHODS: Serum (S) and synovial fluid (SF) from 59 patients with RA, and a knee joint effusion and serum from 90 control subjects were studied with radioimmunoassays for the aminoterminal propeptides of type I and type III procollagens (PINP and PIIINP, respectively). The breakdown of type I collagen was quantified with a radioimmunoassay for the cross linked carboxyterminal telopeptide of type I collagen (ICTP). RESULTS: About 50% of the patients had increased S-ICTP and S-PIIINP values, whereas S PINP was increased in only 20% of the patients. The mean SF:S ratios of these markers varied between 4 (for ICTP) and 340 (for PIIINP), indicating that markers of collagen metabolism are formed locally and then released into the circulation. SF-PINP and SF-PIIINP correlated with each other (rs = 0.86, p < 0.001) and with SF-ICTP (rs = 0.69, p < 0.001, and rs = 0.65, p < 0.001, respectively). SF-ICTP was clearly related to radiographic findings in the corresponding knee joint, patients with gross bone deformation having the greatest SF-ICTP concentrations. S-ICTP and S-PIIINP also correlated with conventional markers of disease activity, such as C reactive protein and joint swelling score. CONCLUSION: Markers of collagen metabolism both in serum and synovial fluid can be measured to provide an assessment of disease process in patients with RA. ICTP and PIIINP are the most informative. PMID- 7492238 TI - Distribution of cases of systemic lupus erythematosus at time of first symptom in an urban area. AB - OBJECTIVES: To determine the geographical distribution of cases of systemic lupus erythematosus (SLE) in a defined geographical area in the East Midlands, UK, and, in particular, to search for spatial variation in cases that may implicate the role of environmental factors in SLE aetiology. METHODS: Six methods of case ascertainment were used. The postcode of the patient's domicile at time of first definite symptom of SLE was used for analysis which included case mapping, probability mapping by electoral ward, and variogram analysis. RESULTS: The study area population of 613,700 contained 200 SLE patients, 188 of whom experienced their first symptom whilst residing in the area. Case mapping revealed 12 SLE patients residing within an area of one square mile, including four men and six patients with RNP antibodies. The use of probability mapping showed five wards in close proximity to each other to have a greater number of SLE cases than would be expected by chance (p < 0.1). The 'cluster' of patients seen on the case map fell into two wards which showed a significant excess of cases only when combined (p = 0.006). The variogram of the incidence rates for each ward did not confirm any structure or pattern to the distribution of cases for the whole area. CONCLUSIONS: Some areas have a greater than expected prevalence of SLE. The normal result from variogram analysis suggests that the cause(s) for these excess number of cases does not have an effect across the whole study area. PMID- 7492240 TI - Enhanced membrane expression of the 52 kDa Ro(SS-A) and La(SS-B) antigens by human keratinocytes induced by TNF alpha. AB - OBJECTIVE: To investigate the membrane expression of the 52 kDa Ro(SS-A) and La(SS-B) antigens in human keratinocytes under the influence of an important mediator of inflammation, TNF alpha. METHODS: Keratinocytes, isolated from human skins obtained at circumcision and identified using monoclonal antibodies, were treated with tumour necrosis factor alpha (TNF alpha) and incubated with antibodies to 52 kDa Ro(SS-A) isolated and purified from patients with systemic lupus erythematosus or Sjogren's syndrome, with mouse monoclonal antibody to La(SS-B), and (as controls) with sera from normal healthy blood donors and a mouse monoclonal antibody to U1RNP 68 kDa. Membrane expression of the 52 kDa Ro(SS-A) and La(SS-B) antigens was detected using cyto enzyme linked immunosorbent assays (ELISAs), laser scanning microscopy, and indirect immunofluorescence. RESULTS: After the incubation with TNF alpha, cyto ELISA revealed a significantly increased membrane binding of 52 kDa Ro(SS-A) antibodies, with a maximum after two hours, followed by enhanced 52 kDa Ro(SS-A) expression during the subsequent 24 hours. The La(SS-B) antigen was expressed rapidly after TNF alpha treatment (within one hour), with a fast decrease to the preincubation value within three hours. Indirect immunofluorescence with fixed normal human keratinocytes confirmed increased 52 kDa Ro(SS-A) and La(SS-B) antigen expression after the incubation with TNF alpha. CONCLUSIONS: TNF alpha mediates 52 kDa Ro(SS-A) and La(SS-B) autoantigen surface expression on human keratinocytes, and may be an important factor both in antibody induction and in the initiation of immunopathogenic processes which occur after antibody binding in autoimmune dermatitis. PMID- 7492239 TI - Mast cells, cytokines, and metalloproteinases at the rheumatoid lesion: dual immunolocalisation studies. AB - OBJECTIVES: To examine the distribution and activation of mast cells (MCs) in the rheumatoid lesion (cartilage-pannus junctions demonstrating cartilage erosion), and to determine whether or not their tissue distribution is related to that for tumour necrosis factor alpha (TNF alpha), interleukin-1 (IL-1), stromelysin-1, and collagenase. METHODS: Immunolocalisation of MC-tryptase was used to identify MCs and their states of degranulation in 35 specimens of cartilage-pannus junctions. Dual immunolocalisation techniques using alkaline phosphatase and peroxidase conjugated antibodies were used to compare the distributions of MCs with the proinflammatory cytokines TNF alpha and IL-1, and the cartilage or matrix degrading enzymes stromelysin-1 and collagenase. RESULTS: Stromelysin-1, TNF alpha, and IL-1 beta were especially prominent at the cartilage-pannus junctions, albeit with patchy distributions. Extracellular MC tryptase, indicative of MC activation/degranulation, was commonly observed at sites of cartilage erosion, and was often associated with the microenvironmental expression of TNF alpha, IL-1 beta, stromelysin-1, and collagenase. Such observations were often associated with localised sites of tissue oedema and stromal disruption. CONCLUSION: MC activation was frequently associated with proinflammatory cytokine and metalloproteinase expression by neighbouring cells, thereby suggesting an important contributory role for the MC in mediating matrix degradation and oedematous changes within microfoci of the rheumatoid lesion. PMID- 7492241 TI - Human blood and synovial fluid neutrophils cultured in vitro undergo programmed cell death which is promoted by the addition of synovial fluid. AB - OBJECTIVE: To assess the influence of inflammatory synovial fluid (SF) on apoptosis of joint and blood neutrophils with particular reference to levels of colony stimulating factors (CSF) contained therein. METHODS: Neutrophils were separated from fresh synovial fluid and from peripheral blood by density gradient centrifugation. Apoptosis was assayed by light microscope morphology and DNA degradation. CSFs were assayed using bone marrow bioassay and enzyme linked immunosorbent assays for granulocyte (G-) and granulocyte macrophage (GM-) CSF. Separated neutrophils were cultured in vitro and exposed to: varying concentrations of SF in which CSF levels were measured, recombinant G-CSF and GM CSF, and hyaluronic acid control solutions. Numbers of apoptotic neutrophils and CSF levels were also measured in fresh SF samples. RESULTS: The addition of autologous or heterologous inflammatory SF to blood or joint cavity neutrophils cultured in vitro caused a significant dose dependent increase in the percentage of cells becoming apoptotic with time as measured morphologically and confirmed by DNA degradation. The effect bore no relationship to levels of CSF in joint fluid, despite our finding that GM-CSF produced inhibition of neutrophil apoptosis in vitro. CONCLUSION: These data suggest that SF contains a factor or factors capable of directly or indirectly promoting neutrophil apoptosis and normally powerful enough to overcome the apoptosis inhibiting effects of cytokines such as GM-CSF at concentrations usually found in inflammatory synovial fluids. PMID- 7492243 TI - The semeiology of arthritis: discriminating between patients on the basis of their symptoms. AB - OBJECTIVES: To examine the intended meaning of words used by patients to describe arthritic symptoms, and to distinguish between different patient groups on the basis of these words. METHODS: A Joint Symptom Questionnaire, developed to resemble the McGill Pain Questionnaire, was given to health professionals (n = 50) and patients with rheumatoid arthritis (RA) (n = 100), fibromyalgia (FM) (n = 50), ankylosing spondylitis (AS) (n = 50), and osteoarthritis (OA) (n = 50). Respondents were invited to define each word by selecting an appropriate heading. Comparison of patient groups was based on the selection of words they chose to describe their joint symptoms. RESULTS: Between health professionals and patients there were no semantic differences in the words given. Patients with FM chose more words to describe their symptoms than the other patient groups (RA median nine words; AS nine words; OA 10 words; FM 12 words). Using receiver operating characteristic curves, a clear distinction between patients with RA, FM, and AS was found, but patients with RA were not readily separated from patients with OA. CONCLUSIONS: There appears to be no semeiological confusion between health professionals and patients regarding arthritic symptoms. However, the spectrum of words chosen by patients to describe the feelings in their joints permits a separation between patients with RA, AS, and FM. Using this questionnaire, patients with RA and OA are symptomatically similar. PMID- 7492242 TI - Differences in oxidative response of subpopulations of neutrophils from healthy subjects and patients with rheumatoid arthritis. AB - OBJECTIVES: To determine whether blood neutrophils from healthy individuals and blood and synovial fluid neutrophils from patients with rheumatoid arthritis (RA) responded differently to priming agonists and stimuli of the oxidative burst and, if so, whether this was a property of a subpopulation of neutrophils. METHODS: Continuous flow electrophoresis was used to separate neutrophils into subpopulations based upon quantitative differences in net negative surface charge. The generation of superoxide anion (O2-) was used as a measure of oxidative activity using 10(-7) mol/l N-formyl-methionylleucyl-phenylalanine (FMLP) as the stimulating agonist and 10(-8) mol/l platelet activating factor (PAF) as the priming agent. RESULTS: The production of O2- by blood and synovial fluid neutrophils from RA patients in response to FMLP was greater than that observed with control blood neutrophils (p < 0.001). Priming of normal blood neutrophils with PAF increased their FMLP induced oxidative burst (p < 0.001), but PAF treatment had no effect on rheumatoid neutrophils. Neutrophils from synovial fluid of RA patients were less electronegative than paired blood samples and exposure of blood neutrophils to FMLP but not PAF reduced their surface charge. Continuous flow electrophoresis isolated three neutrophil subpopulations: cells of least surface electronegativity were ascribed to pool P1 and cells of greatest surface electro-negativity to P3. Normal blood neutrophils from P3, but not P1, showed increased oxidative activity after PAF priming (twofold increase; p < 0.01), whereas the responsiveness of rheumatoid blood and synovial fluid neutrophils from P1 and P3 was not modified by PAF treatment under the same conditions. CONCLUSION: It is suggested that most of the circulating neutrophils in RA are already in a state of readiness to generate O2- upon activation by an inflammatory stimulus. This is in contrast to normal blood neutrophils, which have both responsive and non-responsive subpopulations with respect to priming agonists. PMID- 7492244 TI - Isolated nail fold vasculitis in rheumatoid arthritis. AB - OBJECTIVES: To establish the clinical significance of isolated nail fold vasculitis in patients with rheumatoid arthritis (RA). METHODS: Patients attending hospital with established RA were assessed by a single observer using unaided vision. Isolated nail fold vasculitis (NFV) was diagnosed as small nail edge or nail fold lesions without any evidence of other extra-articular or systemic vasculitis. Patients were followed prospectively. RESULTS: Thirty patients with isolated NFV were identified and followed for a median interval of 22 months. Three patients died within three months of diagnosis and six developed complications possibly indicative of extra-articular disease: xerostomia (three), pericarditis (one), pleural effusion and empyaema (one), pulmonary fibrosis (one). No patient developed systemic vasculitis. CONCLUSION: Isolated NFV has a favourable prognosis compared with systemic vasculitis. There is a low risk of developing systemic or extra-articular disease. PMID- 7492246 TI - Brown's syndrome: an important cause of diplopia in systemic lupus erythematosus. PMID- 7492245 TI - Interleukin-1 expression by neutrophils in rheumatoid arthritis. AB - OBJECTIVE: To determine if neutrophils from blood and synovial fluid of patients with rheumatoid arthritis and other joint arthropathies express interleukin-1 beta mRNA. METHODS: RNA was isolated from neutrophils from patient and control blood, and synovial fluid of patients, probed in northern blots, and quantified by densitometry. It was also isolated and analysed from control blood neutrophils after incubation in vitro with granulocyte macrophage colony stimulating factor (GM-CSF). RESULTS: Neutrophils from the synovial fluid of patients with rheumatoid arthritis contained low levels of mRNA for interleukin-1 beta--between 0.1 and 2% of those observed during stimulation of control neutrophils with GM CSF for one hour. Higher levels (4-40% of the maximal GM-CSF values) were observed in blood neutrophils from patients with rheumatoid arthritis. CONCLUSIONS: Neutrophils contribute to the cytokine network in rheumatoid arthritis. In some circumstances, activation of transcription may occur within the circulation of these patients. PMID- 7492247 TI - Mechanism of haematuria in lupus nephritis. PMID- 7492248 TI - Successful application of high dose intravenous immunoglobulins in Sjogren's syndrome associated arthritis. PMID- 7492249 TI - Asymmetrical nodular osteoarthritis in a patient with a hemiparesis. PMID- 7492250 TI - Sweet's syndrome associated with undifferentiated connective tissue syndrome. PMID- 7492251 TI - The NMDA receptor as a site for psychopathology. Primary or secondary role? PMID- 7492252 TI - Constraints on concepts of pathogenesis. Language and the speciation process as the key to the etiology of schizophrenia. PMID- 7492253 TI - Altered glutamatergic and GABAergic mechanisms in the cingulate cortex of the schizophrenic brain. PMID- 7492254 TI - The reliability and validity of two structured diagnostic interviews for personality disorders. AB - BACKGROUND: The reliability and validity of Axis II diagnoses were investigated in a sample of 108 patients with nonpsychotic Axis I disorders. METHODS: Patients were assessed for personality disorders (PDs) with either the Personality Disorder Examination (PDE) or the Structured Interview for DSM-III-R Personality (SIDP-R). Validity was examined by comparing interview diagnoses with "best estimate" consensus diagnoses assigned by a panel of judges. RESULTS: Interrater reliabilities were excellent when using continuous data (eg, total or cluster scores; intra-class correlation coefficients, .82 to .92); they were lower with categorical diagnoses (eg, any PD vs no PD; kappa = 0.55 and 0.58 with the two interviews). Validity coefficients (ie, kappa values reflecting agreement between the interviews and the consensus diagnosis) for the decision of any PD vs no PD were 0.18 (56% agreement) with the PDE and 0.37 (75% agreement) with the SIDP-R; validity coefficients for identifying cases of "marked" PD were 0.21 (62% agreement) with the PDE and 0.24 (60% agreement) with the SIDP-R. CONCLUSIONS: There have been important advances in the development of structured interviews for Axis II diagnoses, but the findings suggest a continued need to be thoughtful about their strengths and weaknesses before accepting their results as definitive diagnostic tests. The findings also demonstrated some of the advantages of continuous vs categorical data. PMID- 7492255 TI - Perceived coercion in mental hospital admission. Pressures and process. AB - BACKGROUND: Patients' perceptions of coercion in admission may affect their attitude toward subsequent treatment, including their inclination to adhere to treatment plans. This study looks at the determinants of patients' perceptions of coercion. METHODS: A sample of 157 patients admitted to a rural Virginia state hospital and a Pennsylvania community hospital were interviewed within 48 hours of admission about their experience of coming to the hospital. All subjects were 17 years or older. Diagnoses were diverse, and 42% were involuntarily committed. The interview gathered an open-ended description of the admission experience followed by a structured interview that included several measures. RESULTS: Perceptions of being respectfully included in a fair decision-making process ("procedural justice") and legal status were most closely associated with perceived coercion, and a significant relationship was found with perceived negative pressures, ie, force and threats. However, only procedural justice was related to the perception of coercion at both sites and with both voluntary and involuntary patients. CONCLUSIONS: Patients' feelings of being coerced concerning admission appears to be closely related to their sense of procedural justice. It may be that clinicians can minimize the experience of coercion even among those legally committed by attending more closely to procedural justice issues. PMID- 7492256 TI - Signs of defensive control locate conflicted topics in discourse. AB - BACKGROUND: Psychodynamic clinicians cite defensive actions observed in evaluation and treatment as a source of important information. Empirical support for such assertions has seldom been based on objective study of recorded psychotherapy. A quantitative study of the association of signs of defensive control with disclosure of conflicted beliefs was undertaken. METHODS: Brief dynamic psychotherapies of two patients with pathological grief reactions were recorded and reviewed by independent judges. Periods of heightened defensive control of verbal and nonverbal communication were quantified using operational definitions. Reliably scored defensive episodes were algorithmically selected by computer programs. The people and topics discussed by patients were independently scored. RESULTS: Highly defensive periods contained disproportionately more frequent instances of patient discourse about people involved in conflict and unresolved topics. CONCLUSIONS: The findings support the psychodynamic hypothesis that observation of defensive behaviors may locate times of communication about conflicted themes. PMID- 7492257 TI - Posttraumatic stress disorder in the National Comorbidity Survey. AB - BACKGROUND: Data were obtained on the general population epidemiology of DSM-III R posttraumatic stress disorder (PTSD), including information on estimated life time prevalence, the kinds of traumas most often associated with PTSD, sociodemographic correlates, the comorbidity of PTSD with other lifetime psychiatric disorders, and the duration of an index episode. METHODS: Modified versions of the DSM-III-R PTSD module from the Diagnostic Interview Schedule and of the Composite International Diagnostic Interview were administered to a representative national sample of 5877 persons aged 15 to 54 years in the part II subsample of the National Comorbidity Survey. RESULTS: The estimated lifetime prevalence of PTSD is 7.8%. Prevalence is elevated among women and the previously married. The traumas most commonly associated with PTSD are combat exposure and witnessing among men and rape and sexual molestation among women. Posttraumatic stress disorder is strongly comorbid with other lifetime DSM-III-R disorders. Survival analysis shows that more than one third of people with an index episode of PTSD fail to recover even after many years. CONCLUSIONS: Posttraumatic stress disorder is more prevalent than previously believed, and is often persistent. Progress in estimating age-at-onset distributions, cohort effects, and the conditional probabilities of PTSD from different types of trauma will require future epidemiologic studies to assess PTSD for all lifetime traumas rather than for only a small number of retrospectively reported "most serious" traumas. PMID- 7492258 TI - Frontal and temporal lobe brain volumes in schizophrenia. Relationship to symptoms and clinical subtype. AB - BACKGROUND: Quantitative magnetic resonance imaging (MRI) studies demonstrate reduced brain volumes in schizophrenics, but specific structural abnormalities have not been clearly delineated. The structural abnormalities of this disorder are likely to be heterogeneous, consistent with its diverse clinical presentation. To investigate the relationship between structural abnormality and clinical symptoms, we examined regional brain and cerebral spinal fluid (CSF) volumes in a large sample of schizophrenic patients and controls, with patients aggregated into clinical subtypes. METHODS: Right and left hemisphere frontal and temporal lobe brain and CSF volumes were quantified from 5-mm axial spin-echo MRIs for 71 schizophrenic patients and 77 age- and sex-matched controls. The following four standardized rating scales were used to assess symptom severity: Negative Symptoms, Disorganization, Schneiderian Delusions and Hallucinations, and Suspicion-Hostility. Patients were also subtyped as either deficit or nondeficit on the basis of enduring negative symptoms. RESULTS: Schizophrenic patients overall exhibited abnormal brain asymmetry, with selective decrease in brain volume in the left temporal and right frontal regions. Left temporal lobe parenchymal volume reduction and CSF volume increase were correlated with the severity of negative symptoms. Consistent with this, the subtype analysis revealed abnormal temporal lobe asymmetry for the deficit subgroup only. Right frontal lobe volume reduction correlated with the duration of illness, independent of symptom severity or schizophrenic subtype. CONCLUSIONS: Abnormal lateral asymmetry suggests selective structural deficits in schizophrenia, rather than diffusely undifferentiated CNS abnormalities. The pattern of regional abnormalities is related to clinical symptoms, with negative symptoms being associated with left temporal lobe rather than frontal lobe abnormality. This is consistent with suggestions of a temporolimbic prefrontal network abnormality in schizophrenia. Further longitudinal studies are warranted, using higher resolution MRI technology and gray matter-white matter segmentation to confirm and extend these findings. PMID- 7492259 TI - Conversions from clozapine to other antipsychotic drugs. PMID- 7492260 TI - Glutamate receptor dysfunction and schizophrenia. AB - In this article, we advance a unified hypothesis pertaining to combined dysfunction of dopamine and N-methyl-D-aspartate glutamate receptors that highlights N-methyl-D-aspartate receptor hypofunction as a key mechanism that can help explain major clinical and pathophysiological aspects of schizophrenia. The following fundamental features of schizophrenia are accommodated by this hypothesis: (1) the occurrence of structural brain changes during early development that have the potential for producing subsequent clinical manifestations of schizophrenia, (2) a quiescent period in infancy and adolescence before clinical manifestations are expressed, (3) onset in early adulthood of psychotic symptoms, (4) involvement of dopamine (D2) receptors in some cases but not others that would explain why some but not all patients are responsive to typical neuroleptic therapy, and (5) ongoing neurodegenerative changes and cognitive deterioration in some patients. We propose that since N methyl-D-aspartate receptor hypofunction can cause psychosis in humans and corticolimbic neurodegenerative changes in the rat brain, and since these changes are prevented by certain antipsychotic drugs, including atypical neuroleptic agents (clozapine, olanzapine, fluperlapine), a better understanding of the N methyl-D-aspartate receptor hypofunction mechanism and ways of preventing its neurodegenerative consequences in the rat brain may lead to improved pharmacotherapy in schizophrenia. PMID- 7492261 TI - 4,5-Dihydro-3-(2-pyrazinyl)naphtho[1,2-c]pyrazole: a potent and selective inhibitor of steroid-17 alpha-hydroxylase-C17,20-lyase (P450 17). PMID- 7492262 TI - Construction of a detailed serotoninergic 5-HT2a receptor model. AB - We have been able to show that the most important 5-HT2a antagonists and agonists, belonging to chemically diverse classes can be fitted accurately into a common pharmacophoric pattern. In this paper we make use of the developed pharmacophore models to construct a fragmental amino acid model reflecting binding properties and affinity data of the structures under study. The proposed fragment model is then superimposed onto the transmembraneous part of the 5-HT2a receptor. For this purpose the helix coordinates from the known structure of bacteriorhodopsin serve as a matrix. New structures with high affinity for the 5 HT2a sites were designed and their biological activities were predicted on the basis of interaction energies calculated with the fragment model. The predicted data show excellent agreement with experimental binding affinities. PMID- 7492263 TI - Preparation and in vitro pharmacology of 5-HT4 receptor ligands. Partial agonism and antagonism of metoclopramide analogous benzoic esters. AB - Alicyclic ester analogues of the gastroprokinetic benzamide metoclopramide (1) and its ester congener SDZ 205557 (2), a 5-HT4 receptor antagonist, were prepared by O-alkylation of 4-amino-5-chloro-2-methoxybenzoate with N-(2-chloroethyl) substituted alicyclic amines. The bromo and iodo analogue of compound 13b (2-(1 piperidinyl)ethyl 4-amino-5-chloro-2-methoxybenzoate) were obtained by halogenation of dechloro-13b with N-halogenated succinimides. The series was evaluated in functional in vitro assays with regard to affinity for serotoninergic 5-HT4, 5-HT3 and muscarinic M3 receptors. The affinities for 5-HT3 and M3 receptors were below 6.0 (pKB or pA2). On 5-HT4 receptors in guinea-pig ileal longitudinal muscle and rat oesophagus, the majority of compounds revealed partial 5-HT4 receptor agonism susceptible to blockade by SDZ 205557, a reference 5-HT4 receptor antagonist (pKB = 7.25-7.73 (guinea-pig ileum) and 7.09-7.43 (rat oesophagus)). The relative agonist potency was in the range of 5-303% (5-HT: 100%). Compound 13b and its bromo analogue 17 were the most potent esters of the series. The enantiomers of 13g ((R)- and (S)-2-(2-methyl-1-piperidinyl)ethyl 4 amino-5-chloro-2-methoxybenzoate) interacted stereoselectively with 5-HT4 receptors and displayed enantiomeric potency ratios (R)/(S) of 4.3-8.7. There was an excellent correlation between (a) antagonist affinity on guinea-pig ileum and rat oesophagus, (b) relative agonist potency on guinea-pig ileum and rat oesophagus, and (c) between antagonist affinity and relative agonist potency within each assay (r2 > 0.91). The new compounds may serve as academic tools in evaluating the functional role of 5-HT4 receptors. The selective partial 5-HT4 receptor agonists presented in this paper may be useful to restore physiological motility and secretion in the gut with reduced or absent propensity to elicit tachycardia and desensitization of the intestinal target receptor. PMID- 7492264 TI - Third generation antitumor platinum(II) complexes of the [1 (fluoro/difluorophenyl)-2-phenylethylenediamine]platinum(II) type. AB - The diastereomeric 1-(fluoro/difluorophenyl)-2-phenylethylene-diamines (4-fluoro: erythro-1/threo-1; 2,4-difluoro: erythro-2/-threo-2; 2,6-difluoro: erythro 3/threo-3) and the diastereomeric 1-(4-fluorophenyl)-2-(3 hydroxyphenyl)ethylenediamines (erythro-4/-threo-4) were synthesized from appropriately substituted stilbenes by reaction with IN3 and subsequent LiAlH4 reduction. Coordination of the 1,2-diphenylethylenediamines to platinum was carried out by use of K2PtI4. The water-soluble aquasulfato-platinum(II) complexes (erythro/threo-1-PtSO4-erythro/threo-4-PtSO4) were obtained from the diiodoplatinum(II) complexes by reaction with Ag2SO4. Additionally erythro/threo 1-PtSO4 and erythro/threo-4-PtSO4 were transformed into the dichloroplatinum(II) complexes (erythro/threo-1-PtCl2, erythro/threo-4-PtCl2) by treatment with KCl. In contrast to the less effective erythro-configurated sulfatoplatinum(II) complexes the threo-analogues showed comparable or even superior activities to cisplatin on the human MDA-MB-231 breast cancer cell line. On the MXT-M-3.2 breast cancer of the mouse only erythro- and threo-4-PtSO4 caused similar effects like cisplatin. The strong inhibitory effect of the diastereomeric sulfatoplatinum(II) complexes on the P-388 leukemia of the mouse was equal to that of cisplatin. On the latter tumor threo-4-PtCl2 was the most active among the less toxic dichloroplatinum(II) derivatives. PMID- 7492266 TI - Synthesis, pharmacological investigation and computational studies on a tricyclic ergoline analog with selective dopamine autoreceptor activity. AB - The novel aminobenzindolone 8 was prepared and evaluated as a potential antipsychotic agent. The target compound was synthesized in eight steps starting from the tetrahydrobenzindolone 9. The key step of the synthesis was an electrophilic amination of the aromatic ketone 11 followed by reductive degradation when the diethoxymethyl group was employed for protection of the lactam nitrogen and also for the benzylic position 2a. Dopamine and serotonin receptor binding studies revealed 8 to be a potent and selective ligand at the D 2 autoreceptor (ki = 4.0 nM). Further in vivo studies including the GBL-test and locomotor activity measurements indicated agonistic activity of 8 at the prejunctional binding sites. Comparison of ab initio based molecular electrostatic isopotential maps corroborates our hypothesis that the dopamine structure 6, containing an intramolecular hydrogen bond donating effect of the meta-HO-group, represents the conformation which is active at the dopamine D-2 autoreceptor. PMID- 7492265 TI - Structure-activity relationship studies of CNS agents, Part 23: N-(3 phenylpropyl)- and N-[(E)-cinnamyl]-1,2,3,4-tetrahydroisoquinoline mimic 1 phenylpiperazine at 5-HT1A receptors. AB - The 5-HT1A receptor affinities and ionization constants of a set of 1 arylpiperazine (4) 1,2,3,4-tetrahydroisoquinoline (6), and -quinoline (7) containing N-(omega-arylalkyl) or N-(E)-cinnamyl substituents as well as two morpholine derivatives (8a, 8b) were determined. It was shown that some tetrahydroisoquinoline (6c, 6d) and morpholine (8a) derivatives were 5-HT1A ligands equipotentto 1-phenylpiperazine (4a) and 1,2,3,4,4a,5-hexahydropyrazino [1,2-a]indole (5). On the basis of molecular modelling studies it was also demonstrated that 6c, 6d and 8a mimicked very well the reference structures of 4a and its rigid analog 5. Another, more complex 1,2,3,4-tetrahydroisoquinoline derivative 3, which served as a model compound to confirm the previously reported 5-HT1A binding mode of derivatives 1a-d and 2, had the highest 5-HT1A affinity (Ki = 6.7 +/- 0.5 nM) of all the investigated compounds. PMID- 7492267 TI - Synthesis, toxicological, pharmacological, and bronchodilating activity in vitro of some xanthineacetic acid derivatives. AB - The possibility of the preparation of some ester derivatives of dimethylxanthines from 1-theobromine- and 7-theophylline acetic acids and 7-(2-hydroxyethyl) theophylline by DCC/DMAP-mediated esterification under mild conditions was studied. The structures of the compounds synthesized and by products isolated were demonstrated by microanalyses, UV-, IR-, and 1H NMR data. Acute toxicity assessment of the compounds on mice showed that compounds 4, 5, 6, and 7 are less toxic than aminophylline. A pharmacological study of the in vitro broncholytic effect (IC50 and pD2 values) of the derivatives and aminophylline showed that the new compound 4 (1,2,3,6-tetrahydro-1,3-dimethyl-2,6-dioxo-7H-purine-7-acetic acid 2-(1,2,3,6-tetrahydro-1,3-dimethyl-2,6-dioxo-7H-purin-7-yl)ethyl ester) has a strong bronchodilating effect on serotonine- and acetylcholine-induced spasm in guinea pig trachea. The same compound does not influence barbiturate-induced hypnosis and locomotor activity, unlike to the effect of the aminophylline, used as a reference substance. PMID- 7492269 TI - Structure-activity relationship studies of CNS agents, Part 22. A search for new trazodone-like antidepressants: synthesis and preliminary receptor binding studies. AB - New 1-phenyl- and 1-(3-chlorophenyl)piperazines containing a 4-[3 (heterocyclic)propyl] fragment were synthesized. It was found that of all the investigated compounds 11b (Ki = 13 +/- 2 nM) and 8b (Ki = 38 +/- 2 nM) were the most active 5-HT1A and 5-HT2A ligands, respectively. Several derivatives (3a, 4a, 8b, 11b, 12b, 13a, and 13b) were selected as good candidates for new, potential antidepressants on the basis of their 5-HT1A/5-HT2A receptor binding profiles. PMID- 7492268 TI - Resolution, EPC-syntheses, absolute stereochemistry, and pharmacology of the (S) (+)- and (R)-(-)-isomers of the MAO-A inhibitor tetrindole hydrochloride. AB - Resolution of (RS)-tetrindole (3) and enantioselective reductions of the imine 7 yielded (S)-(+)-(4) and (R)-(-)-tetrindole (5). The absolute stereochemistry of 4 was established by X-ray analysis of the corresponding Mosher amide 6. From in vitro as well as in vivo data (MAO-inhibition, levels of monoamines and their respective metabolites in rat brain), 4 was identified as the eutomer. PMID- 7492270 TI - Synthesis and dopamine receptor binding studies of homochiral 8-aminopyrido[1,2 a]indoles. AB - Starting from L-aspartic acid the preparation of 8-aminopyrido[1,2-a]indole derivatives as benzo-fused analogs of the dopamine autoreceptor agonist 1 is reported. The key step of the synthesis is the Tf2O induced cyclization of the 1,2-amino alcohol 6. Receptor binding studies indicated selective affinity for the D-2 autoreceptor. Among the tested compounds, the dipropylamino derivative 2 showed the highest affinity for the D-2 receptor labelled with the selective autoreceptor agonist pramipexole (IC50 value: 450 nM). Thus, 2 is 15 times less potent than the aminoindolizine 1. PMID- 7492271 TI - Surgery in China. PMID- 7492273 TI - Liposomes modulate Kupffer cell endotoxin response. AB - OBJECTIVES: To test the hypothesis that pretreatment with liposomes enriched with the omega 3 fatty acid docosahexaenoic acid (22:6 omega 3) will alter the Kupffer's cell and systemic cytokine (tumor necrosis factor and interleukin-6) response to endotoxin challenge, and to demonstrate alterations in Kupffer's cell phospholipid fatty acid composition after in vivo liposome treatment. DESIGN: Nonrandomized controlled laboratory investigation in Wistar rats. INTERVENTIONS: Animals were assigned to three pretreatment groups: no liposomes; liposomes, 100 mg/kg; or liposomes, 400 mg/kg given by bolus intravenous injection with the animals under inhalation anesthesia. Eighteen hours after liposome treatment, each group was challenged with Escherichia coli lipopolysaccharide (3 mg/kg intraperitoneally in 10 mL of lactated Ringer's solution) or lactated Ringer's solution only. In a separate set of experiments, Kupffer's cells were obtained from animals pretreated with liposome, 400 mg/kg, or controls and challenged with lipopolysaccharide (1, 100, or 10(4) ng/mL) in vitro. OUTCOME MEASURES: Serum and Kupffer's cell supernatant tumor necrosis factor and interleukin-6 bioactivity, Kupffer's cell phospholipid fatty acid composition, survival, and liver histologic findings. RESULTS: In vivo liposome pretreatment (400 mg/kg) resulted in significant increases in serum tumor necrosis factor and interleukin 6 levels 90 minutes after intraperitoneal lipopolysaccharide challenge (P < .05 vs no liposomes). Kupffer's cells isolated from liposome-treated animals (400 mg/kg) compared with untreated controls release significantly more tumor necrosis factor and interleukin-6 after lipopolysaccharide stimulation in vitro in a dose dependent response (P < .05). Liposome treatment increased total polyunsaturated fatty acid, total omega 3, and docosahexaenoic acid 22:6 omega 3 content in Kupffer's cell phospholipids compared with untreated controls. Survival 24 hours after lipopolysaccharide challenge was reduced by liposome (400 mg/kg) pretreatment (P < .05 by chi 2 test). Livers from each treatment group demonstrated focal areas of hepatocyte necrosis and inflammatory cells. CONCLUSION: Liposome pretreatment increases the circulating and Kupffer's cell cytokine response to endotoxemia, increases Kupffer's cell polyunsaturated fatty acid content, and is associated with reduced survival. PMID- 7492274 TI - Antimicrobial effects of granulocyte-macrophage colony-stimulating factor in protein-energy malnutrition. AB - OBJECTIVES: To evaluate, in a murine model of protein-energy malnutrition, whether granulocyte-macrophage colony-stimulating factor (GM-CSF) improves the host response to a septic challenge and to determine the potential mechanisms involved. DESIGN: Nonblinded study of GM-CSF in mice with protein-energy malnutrition. SETTING: A university-based surgical laboratory and animal facility. INTERVENTION: In study 1, malnourished mice were randomized to receive either GM-CSF (120 micrograms/kg subcutaneously to receive either GM-CSF (120 micrograms/kg subcutaneously from day 4 to 7 of the protein-free diet) or saline vehicle as a control. On day 7, all mice were given Candida albicans (5 x 10(5) organisms intravenously). In study 2, malnourished mice received the same dose of GM-CSF or saline vehicle for 7 days of the protein-free diet. MAIN OUTCOME MEASURES: In study 1 mice were followed up for survival. In study 2, after 7 days of diets, splenic macrophages were harvested and were assayed for interleukin-6, superoxide anion, and nitric oxide production. Splenocytes were stimulated with concanavalin A (5 micrograms/mL) for interleukin-4, interleukin-10, and interferon-gamma production. RESULTS: Treatment with GM-CSF significantly enhanced survival in malnourished mice infected with C albicans. Treatment with GM-CSF was associated with increased production from splenic macrophages of interleukin-6, superoxide anion, and nitric oxide as well as decreased interleukin-4 production from splenocytes. CONCLUSIONS: This study suggests a beneficial role for GM-CSF in the malnourished host predisposed to infection. The antimicrobial properties of GM-CSF may function through enhanced production of nitric oxide and superoxide anion. PMID- 7492272 TI - Induction of heat shock protein 72 prevents neutrophil-mediated human endothelial cell necrosis. AB - OBJECTIVE: To examine the hypothesis that induction of heat shock proteins in human endothelial cells (ECs) by either heat shock or sodium arsenite could prevent subsequent EC necrosis induced by activated human polymorphonuclear neutrophil leukocytes (PMNs). DESIGN: Cultures of ECs were exposed to heat shock (42 degrees C, 30 to 60 minutes) or sodium arsenite (40 to 320 mumol/L) for 6 hours to induce the expression of a heat shock protein of 72-kd molecular weight (HSP-72). Activated PMNs were subsequently added to these ECs for 24 hours to evaluate the ability of HSP-72 to prevent activated PMN-mediated EC necrosis. RESULTS: Neither EC necrosis nor apoptosis was induced by heat shock. Sodium arsenite (40 to 80 mumol/L) did not induce EC necrosis, although 320-mumol/L sodium arsenite caused a significant increase in EC necrosis. Sodium arsenite (80 to 320 mumol/L) also induced dose-dependent EC apoptosis. Endothelial cells exposed to heat shock and sodium arsenite (40 and 80 mumol/L) significantly attenuated subsequent EC necrosis induced by activated PMNs. However, sodium arsenite at 320 mumol/L aggravated activated PMN-mediated EC necrosis. Expression of HSP-72 was detected after ECs were treated both with heat shock and sodium arsenite (40 to 320 mumol/L) for 6 hours. CONCLUSION: Induction of HSP-72 in ECs by a thermal or nonthermal mechanism could prevent activated PMN-mediated EC necrosis, which may favor increased vascular permeability during systemic inflammatory response syndrome. PMID- 7492275 TI - Both prostaglandin E2 and nitric oxide sequentially mediate the tumor necrosis factor alpha-induced inhibition of surfactant synthesis by human type II pneumocytes. AB - BACKGROUND: Tumor necrosis factor alpha (TNF-alpha)-induced inhibition of surfactant synthesis seems to participate in the pathogenesis of the adult respiratory distress syndrome. OBJECTIVES: To examine the ability of human type II pneumocytes to produce nitric oxide (NO) in the presence of TNF-alpha and, in addition, to explore the role of this radical in the transduction of the cytokine signal. DESIGN: Multiple organ donors were the source of lung tissue specimens. Type II pneumocytes were isolated by enzymatic digestion, adherence separation of macrophages, and gradient purification. After 24-hour preculture, cells were cultured for 24 hours in the presence or absence of TNF-alpha (100 ng/mL), sodium nitroprusside (100 mumol/L), N omega-nitro-L-arginine methyl ester (NAME) (1 mmol/L), methylene blue (10 mumol/L),8-bromo-3',5'-cyclic guanosine monophosphate (8-Br-cGMP) (1 mmol/L), prostaglandin E2 (PGE2) (0.1 mumol/L), indomethacin (30 mumol/L), and combinations. The NO release to the medium and cGMP and PGE2 contents of the cells were measured. RESULTS: The incorporation of 14C-labeled glucose (D-[U-14C]glucose) into phosphatidylcholine and phosphatidylglycerol was selectively inhibited either by 8-Br-cGMP or in the presence of TNF-alpha, PGE2, or nitroprusside, all of which caused an increase in the intracellular levels of cGMP. The inhibitory effect of TNF-alpha was partially reverted by indomethacin, NAME, N-monomethyl arginine, or methylene blue. The inhibitory effect of PGE2 was partially reverted by NAME, while that of nitroprusside was reverted by methylene blue, but not by indomethacin. Tumor necrosis factor alpha induced an increase in PGE2 (4.31 +/- 0.27 vs 1.65 +/- 0.17-pg/microgram protein, n = 10, P < .01) and cGMP (0.238 +/- 0.012 vs 0.109 +/- 0.014-pmol/microgram protein, n = 10, P < .01) cell content and in the NO release to the medium (3.10 +/- 0.14 vs 1.19 +/- 0.11 nmol/microgram protein, n = 10, P < .01). The basal NO release to the medium was also increased in the presence of PGE2. The NAME, which blocked NO generation and cGMP increase, did not affect PGE2 production in response to TNF-alpha. However, indomethacin, which blocked PGE2 production, also blunted NO generation and cGMP increase. CONCLUSIONS: The NO generation, secondary to PGE2 production, seems responsible for the TNF-alpha-induced inhibition of phosphatidylcholine synthesis by human type II pneumocytes. Nitric oxide seems to exert this effect through activation of guanylyl cyclase. PMID- 7492277 TI - Polymyxin B prevents increased sympathetic activity and alveolar macrophage tumor necrosis factor release in parenterally fed rats. AB - OBJECTIVE: To determine the effects of polymyxin B sulfate in rats fed by total parenteral nutrition on norepinephrine excretion, macrophage tumor necrosis factor production, and bacterial translocation. DESIGN: Randomized animal study. SETTING: A university teaching hospital in Seattle, Wash. MATERIALS AND METHODS: Three groups of rats were studied: chow plus intravenous saline, total parenteral nutrition, or total parenteral nutrition supplemented with polymyxin B sulfate. After 5 days, urinary excretion of norepinephrine and epinephrine was calculated, peritoneal and alveolar macrophages were cultured, and their spontaneous and lipopolysaccharide-stimulated tumor necrosis factor production was measured. Mesenteric lymph nodes were cultured for bacteria. RESULTS: Rats fed by total parenteral nutrition had increased urine norepinephrine excretion (33%) and alveolar macrophage tumor necrosis factor production (80%) and trends for increased epinephrine excretion and bacterial translocation compared with rats fed chow. Alveolar but not peritoneal macrophage tumor necrosis factor production was significantly related to norepinephrine excretion (r = .5, P < .01). The addition of polymyxin B to total parenteral nutrition decreased weight gain (P < .05), urinary norepinephrine excretion (P < .01), and alveolar macrophage tumor necrosis factor production (P < .05) compared with rats fed by total parenteral nutrition. Polymyxin B also tended to decrease the magnitude of bacterial translocation. CONCLUSIONS: Alveolar macrophage tumor necrosis factor production appears to be influenced by sympathetic nervous activity. Total parenteral nutrition-induced endotoxemia may indirectly alter macrophage function by stimulating sympathetic nervous activity. PMID- 7492276 TI - Evidence for a cyclic guanosine monophosphate-dependent, carbon monoxide mediated, signaling system in the regulation of TNF-alpha production by human pulmonary macrophages. AB - BACKGROUND AND OBJECTIVES: The second messenger cyclic guanosine 3',5' monophosphate (cGMP) seems to be implicated in the release of tumor necrosis factor alpha (TNF-alpha) by activated macrophages. There is controversy regarding the potential of human macrophages to produce nitric oxide (NO). Since guanylate cyclase can be activated also by carbon monoxide (CO) and this gas may be formed endogenously, we examined the ability of human pulmonary macrophages to produce CO in the presence of lipopolysaccharide (LPS) or LPS+interferon gamma (IFN gamma). In addition, the source and the relative contribution of this molecule to the LPS-induced increase in cell cGMP content and TNF-alpha release were explored. DESIGN: Interstitial macrophages were obtained from multiple organ donor lungs by enzymatic digestion. After 24-hour preculture, purified macrophages were cultured for 24 hours in the presence or absence of LPS, LPS+IFN gamma, CO (250 and 500 mumol/L), sodium nitroprusside, 8-Br-cGMP, hemoglobin, methylene blue, zinc-protoporphyrin IX, hemin, S-adenosylmethionine, deferoxamine mesylate, or combinations. The cGMP content of the cells and TNF-alpha, CO, and NO release to the medium were determined. RESULTS: In the presence of LPS, TNF alpha production was not accompanied by any detectable increase in the NO release to the medium. However, an increase in medium CO concentration (mean +/- SEM) (5.81 +/- 0.20 vs 3.74 +/- 0.08 pmol/microgram protein; n = 11; P < .01) and cell cGMP content (0.273 +/- 0.021 vs 0.138 +/- 0.019 pmol/microgram protein; n = 10; P < .01) was observed. These changes were more pronounced in the presence of LPS+IFN-gamma. Release of TNF-alpha also was induced by both sodium nitroprusside and 8-Br-cGMP. In contrast, methylene blue, a guanylate cyclase inhibitor, inhibited LPS-, LPS+IFN-gamma-, and sodium nitroprusside-induced TNF-alpha production and cGMP increase; hemoglobin, which traps CO, had a similar effect. CONCLUSION: Intracellular cGMP increase, secondary to an endogenous production of CO, participates in the release of TNF-alpha by activated human pulmonary macrophages. PMID- 7492279 TI - Primary vs secondary iliopsoas abscess. Presentation, microbiology, and treatment. AB - OBJECTIVE: To review the characteristics of patient presentation, microbiology, and treatment of primary iliopsoas abscess. DESIGN: A case series of patients with iliopsoas abscess diagnosed on computed tomographic scans from 1987 to 1994. SETTING: Tertiary care inner-city university hospital. PATIENTS: Eleven patients with secondary iliopsoas abscess, defined as being secondary to gastrointestinal or genitourinary causes or trauma, and seven patients with primary abscess, defined as the absence of the above causes. MAIN OUTCOME MEASURES: Patient characteristics, presenting symptoms and signs, microbiologic characteristics, treatment, and clinical course of patients with primary iliopsoas abscesses compared with those in patients with secondary abscesses. RESULTS: In the primary group, six patients (86%) were intravenous drug users and four (57%) were positive for human immunodeficiency virus. Staphylococcus aureus grew from cultures from five of seven patients with primary abscesses, whereas secondary abscesses had enteric flora. The typical patient presentation included fever, with complaints of pain in the flank, hip, or abdomen. Comparison of abscess drainage options showed shorter hospitalizations for surgical drainage than for percutaneous drainage (15.9 vs 28.5 days; P < or = .01). CONCLUSIONS: A patient who presents with pain in the flank, hip, or abdomen may have a primary iliopsoas abscess. Computed tomography is the standard method of diagnosis. Antibiotic regimens for patients with primary iliopsoas abscess should include coverage for S aureus, and patients with secondary abscesses should have antibiotic regimens tailored for enteric bacteria. Drainage of abscess is essential for appropriate treatment, and surgical drainage is superior to percutaneous drainage in achieving prompt recovery. PMID- 7492278 TI - Differential expression of intestinal and splenic cytokines after parenteral nutrition. AB - OBJECTIVE: To determine the effect of parenteral nutrition (PN) on the expression of message for inflammatory cytokines in the spleen and different segments of the intestine. DESIGN: Randomized controlled trial. PARTICIPANTS: Eleven adult male Sprague-Dawley rats weighing 250 to 300 g. INTERVENTIONS: All rats underwent central venous cannulation and were randomized to two groups. Group 1 (n = 6) received saline solution infusion and chow ad libitum; group 2 (n = 5) received lipid-free PN with no oral feeding. After 7 days, the animals were killed and the spleens and segments of small and large intestine were removed. MAIN OUTCOME MEASURES: The expression of message for tumor necrosis factor alpha (TNF-alpha), interleukin-6 (IL-6), and IL-1 in the spleen and intestine was determined using a semiquantitative reverse transcription polymerase reaction. Splenic macrophages were isolated and cultured for 24 hours with and without lipopolysaccharide. Production of TNF-alpha and IL-6 was determined by bioassay followed by enzyme linked immunosorbent assay. RESULTS: After 7 days of infusion, messenger RNA (mRNA) expression for TNF-alpha, IL-1, and IL-6 was increased in the jejunum (P < .05), and TNF-alpha mRNA and IL-6 mRNA expression was decreased in the spleen (P < .01) of PN-fed animals when compared with saline/chow controls. In addition, TNF-alpha mRNA expression was increased in the cecum (P < .05), IL-1 mRNA expression was increased in the ileum (P < .05), and IL-6 mRNA expression was increased in the cecum (P < .05) and Peyer's patches (P < .007) in the PN-fed animals. Production of TNF-alpha and IL-6 by splenic macrophages was decreased following PN infusion in both lipopolysaccharide-treated and untreated cultures (P < .05). CONCLUSIONS: Infusion of lipid-free PN induces a differential mRNA expression for inflammatory cytokines in the spleen and intestine with an overall up-regulation of the expression of inflammatory cytokines in the intestine and a down-regulation in the spleen. These data provide evidence that the regulatory mechanisms for cytokine production are different in the intestine and the spleen. Further study is needed to elaborate the mechanism of this differential expression following lipid-free PN infusion. PMID- 7492280 TI - Inflammatory response in peritoneal exudate and plasma of patients undergoing planned relaparotomy for severe secondary peritonitis. AB - OBJECTIVE: To study the pattern of intraperitoneal cytokine release in secondary peritonitis and its correlation with plasma levels and prognosis. DESIGN: Noncomparative descriptive case series. SETTING: Department of surgery in a university hospital. PATIENTS: Seventeen consecutive patients undergoing planned relaparotomy for severe intra-abdominal infection (Acute Physiological and Chronic Health Evaluation [APACHE II] score > 10; mean score, 17.5). INTERVENTIONS: The following were measured at the first and last serial operations in the peritoneal exudate and plasma: endotoxin, tumor necrosis factor alpha (TNF-alpha), interleukin-1 (IL-1), interleukin-6 (IL-6), elastase, and neopterin. MAIN OUTCOME MEASURES: Survival and death. RESULTS: Six patients died. Peritoneal endotoxin levels were significantly higher than in the plasma and were significantly higher in the nonsurvivors. Plasma TNF-alpha, IL-6, elastase, and neopterin levels remained elevated in the nonsurvivors prior to death. Levels of TNF-alpha, IL-6, elastase, and endotoxin were 19, 993, 239, and 7 times higher, respectively, in the peritoneal exudate than in plasma, all significant differences. Elastase and TNF-alpha levels decreased in survivors during the operative treatment but remained elevated in the nonsurvivors. CONCLUSIONS: Secondary peritonitis is associated with a significant cytokine-mediated inflammatory response that is compartmentalized in the peritoneal cavity and indicates an adverse prognosis. Levels of cytokines in the exudate of peritonitis may be used to better stratify the severity of peritonitis and, in future, to guide local therapy. PMID- 7492281 TI - Antibiotic pharmacokinetics following fluid resuscitation from traumatic shock. AB - OBJECTIVE: To describe the pharmacokinetic profile of aztreonam and vancomycin hydrochloride in a clinically relevant experimental model of hemorrhagic shock and trauma. METHODS: Ten mongrel pigs (mean +/- SD weight, 26.7 +/- 6.4 kg) were anesthetized with fentanyl citrate and ventilated, and an indwelling catheter was placed in the jugular vein. On day 3, all pigs were subjected to fentanyl administration, ventilation, soft-tissue injury, and an arterial hemorrhage (mean +/- SD, 40% +/- 8%). After a 1-hour shock period, baseline hemodynamics were restored by reinfusing shed blood plus twice the shed volume as lactated Ringer's solution. Aztreonam and vancomycin were infused on day 1, after resuscitation on day 3, and on days 4 and 8. Serial plasma samples were collected for 6 hours after treatment, and differences were compared with analysis of variance. RESULTS: Aztreonam clearance initially decreased with trauma, but subsequently increased by 48% (P < .02) by day 8. Aztreonam steady-state volume decreased by 34% (P = .05, baseline value vs that on day 8). Vancomycin clearance was increased between 25% and 52% (P < .001) on days 3, 4, and 8 compared with the baseline value. Vancomycin steady-state volume initially increased with trauma (P = .009), but it subsequently decreased by 29% (P < .001) on day 8. These data cannot be explained by changes in plasma volume per se because levels of plasma sodium, potassium, chloride, and calcium were within normal reference ranges at all time points. Neither liver nor renal functions were severely impaired because levels of serum urea nitrogen, bilirubin, liver enzymes, creatinine, and plasma proteins were within normal reference ranges. Furthermore, our previous work demonstrated that systemic and splanchnic organ oxygen delivery and demand were near normal immediately after fluid resuscitation and for at least 3 days thereafter; thus, there were probably no major perfusion abnormalities in the liver or kidney. CONCLUSIONS: For at least 5 days after trauma, clearance and steady-state volume of aztreonam and vancomycin are altered. These changes suggest that the interval and magnitude of dosing should be adjusted, relative to the standard recommended dosages of each antibiotic, to maximize their efficacy. Similar studies should be done for other antibiotics. PMID- 7492282 TI - Interleukin-13 effectively down-regulates the monocyte inflammatory potential during traumatic stress. AB - OBJECTIVES: To determine the potential of interleukin-13 (IL-13) to modify in vitro lipopolysaccharide-induced monocyte-macrophage (MO) activity in human cells from individuals who had sustained either major mechanical or burn injury and to investigate whether the effect of IL-13 is different on MOs that have been preactivated under traumatic stress than on monocytic cells from healthy volunteers. DESIGN: Peripheral MOs from 20 controls and 16 patients after major burn or mechanical trauma were separated on days 1, 3, 5, and 7 after injury and incubated with lipopolysaccharide (1 microgram/mL) in the presence or absence of IL-13 (10 ng/mL) for 4 hours and for 20 hours. Thereafter, the following measures were determined from the culture supernatants: neopterin, nitric oxide, tumor necrosis factor alpha, IL-1 beta, IL-6, and IL-8. RESULTS: Ex vivo lipopolysaccharide-activated MOs, compared with control cells, displayed considerably enhanced inflammatory activity during the immediate posttraumatic course, with a substantial and consistent elevation of levels of tumor necrosis factor alpha and IL-6. The addition of human recombinant IL-13 to the MO cultures resulted in an effective down-regulation of the synthesis of tumor necrosis factor alpha, IL-1 beta, and IL-6 as well as IL-8, showing an average reduction of mediator production to two thirds of the value found in corresponding sole lipopolysaccharide-stimulated cultures. The impact of human recombinant IL-13 on control MOs was almost identical for IL-6 and IL-1 beta, slightly lower for IL-8, and nonexistent for tumor necrosis factor alpha. CONCLUSION: From this study and preexisting findings, we conclude that, based on its biologic properties, IL-13 should be tested as a biologic response modifier for acute states of trauma induced host defense deficiency. PMID- 7492283 TI - Pentoxifylline attenuates pulmonary macromolecular leakage after intestinal ischemia-reperfusion. AB - OBJECTIVE: To assess the effects of pentoxifylline posttreatment on hemodynamic variables and acute pulmonary injury in the rat intestinal ischemia-reperfusion (I-R) model, using a recently developed method of fluorescent intravital pulmonary videomicroscopy. DESIGN: Anesthetized male Sprague-Dawley rats were cannulated for measurement of mean arterial pressure, heart rate, cardiac output, arterial blood gas values, and hematocrit. Rats underwent isolation of the superior mesenteric artery for intestinal I-R (45 minutes of ischemia, 120 minutes of reperfusion) and right lateral thoracotomy for pulmonary videomicroscopy. Epi-illumination fluorescent videomicroscopy was used to quantitate leakage of intravascular fluorescently labeled albumin into alveoli, while hemodynamic variables were simultaneously recorded. In the treatment groups, pentoxifylline was administered after 30 minutes of intestinal ischemia. Data (mean +/- SEM) were recorded before and during intestinal ischemia and after reperfusion at 30-minute intervals. MAIN OUTCOME MEASURE: The appearance of fluorescently labeled albumin into alveolar airspaces was quantitated off-line by computer and reported as the alveolar leak index. RESULTS: Intestinal I-R caused alveolar macromolecular leakage, marked by a 300% +/- 48% increase from baseline (P < .05) in the alveolar leak index. Intestinal I-R also produced systemic hemodynamic instability demonstrated by a decrease in the mean arterial blood pressure (-36% +/- 5% vs baseline, P < .05) and cardiac output (-42% +/- 6% vs baseline, P < .05), metabolic acidosis (final arterial pH of 7.17, P < .05 vs initial pH), and a 2.3-fold increase in the intravenous fluid requirement when compared with that in sham animals (P < .05). Treatment with pentoxifylline 30 minutes after intestinal ischemia attenuated pulmonary macromolecular leakage (P < .05 vs nontreated I-R) and reduced the decrease in cardiac output (-15% +/- 7% vs baseline, not statistically significant). Pentoxifylline treatment had no effect on the mean arterial blood pressure, heart rate, metabolic acidosis, or intravenous fluid requirement. CONCLUSIONS: Pentoxifylline reduces alveolar capillary membrane injury and subsequent protein leakage and improves cardiac output when administered after 30 minutes of intestinal ischemia. These data suggest that pentoxifylline may be a possible candidate as a future therapy for acute pulmonary dysfunction. Further studies in human patients are necessary. PMID- 7492284 TI - Immunosuppression augments growth of graft-adherent Staphylococcus epidermidis. AB - OBJECTIVE: To determine if systemic suppression of host defenses during graft implantation alters the initial adherence and subsequent growth of Staphylococcus epidermidis on vascular prostheses. DESIGN: Dacron grafts 1 cm2 were implanted in the back subcutaneous tissue of Swiss-Webster mice (n = 247), followed by topical inoculation with 2 x 10(7), 2 x 10(5), 2 x 10(3), or 2 x 10(1) colony-forming units of S epidermidis. Half of the mice were immunosuppressed with cyclophosphamide (150 mg/kg intraperitoneally), to achieve a consistent, significant decrease in the white blood cell count and major histocompatibility complex class II (Ia) expression. Control mice received an equal volume of saline solution. Graft bacterial biofilm concentrations were determined at 1 day for adherence and within 2 weeks for bacterial growth, by using sonication and quantitative agar culture. RESULTS: Immunosuppression did not significantly alter the initial adherence of bacteria to vascular grafts. Immunosuppressed animals that were inoculated with 2 x 10(7) and 2 x 10(5) colony-forming units of S epidermidis had significantly higher bacterial biofilm concentrations as compared with those in control animals. Graft infection persisted at 14 days in all animals, with and without immunosuppression. CONCLUSIONS: Suppression of immune function during graft implantation augmented growth of adherent bacteria. The effect of short-term perioperative immunosuppression on late-appearing S epidermidis graft infection needs further study. PMID- 7492285 TI - Migraine during infancy. PMID- 7492286 TI - Morphologic cerebral asymmetries and handedness. PMID- 7492287 TI - Eye movement abnormalities in systemic lupus erythematosus. AB - OBJECTIVE: To describe eye movement abnormalities in patients with systemic lupus erythematosus (SLE). DESIGN: Between January 1, 1970, and June 30, 1995, 113 patients with a clinical diagnosis of SLE were examined. Of these, 33 had ocular motor abnormalities and a diagnosis of SLE as defined by the revised criteria of the American College of Rheumatology. SETTING: Patients hospitalized on the wards of the Los Angeles County-University of Southern California Medical Center, Los Angeles. SUBJECTS: Thirty-three patients, 82% women, ranging in age from 19 to 58 years (mean, 33.5 years). The average duration of SLE was 3.5 years, and 10 patients had been symptomatic for 5 years or longer. RESULTS: Of 55 ocular motor signs, 33 involved limitation of eye movements or abnormal eye position at rest; abnormal spontaneous eye movements occurred 12 times; ptosis was seen in eight patients and psychogenic signs in two. Sixteen patients (48%) had brain-stem infarcts; 11 had other known causes, including three with meningitis, two with hysteria, and one each with ocular myositis, pseudotumor cerebri, both ocular myositis and pseudotumor cerebri, Guillain-Barre syndrome, hyperosmolality, and hypoxic encephalopathy. In four patients with isolated sixth nerve palsies and two with isolated ptosis, the cause remained unknown. CONCLUSIONS: Ocular motor signs in SLE are uncommon and often transitory. When present, they help to ascertain the location, and often the cause, of neurologic involvement. PMID- 7492288 TI - Regional distribution of neurofibrillary tangles and senile plaques in the cerebral cortex of very old patients. AB - OBJECTIVES: To examine the correlations between senile lesion densities and development of dementia symptoms in very old people. To perform a quantitative neuropathologic evaluation of several cortical and subcortical areas in a series of 29 nonagenarians and centenarians. PATIENTS: Ten patients with no cognitive impairment and 19 patients with clinically overt Alzheimer's disease. DESIGN: Neuropathologic case series. Severity of Alzheimer's disease was assessed with the Mini-Mental State examination and by postmortem chart review using the extended Clinical Dementia Rating Scale. Comparisons between neurofibrillary tangle and senile plaque densities in demented and nondemented individuals were performed by analysis of covariance controlling for age at the time of death. SETTING: Studies were conducted at the Psychiatric and Geriatric hospitals of the University of Geneva School of Medicine in Geneva, Switzerland. MAIN OUTCOME MEASURE: Correlations between clinical diagnosis and severity of Alzheimer's disease and neuropathologic change densities. RESULTS: Statistically significant differences were found in neurofibrillary tangle densities in the superior parietal, superior temporal, anterior and posterior cingulate cortex, and nucleus basalis of Meynert between nondemented and Alzheimer's disease cases. The superior parietal and posterior cingulate cortex contained significantly higher senile plaque counts in demented compared with nondemented cases. In contrast to younger demented cases, the number of senile plaques in the neocortex was correlated with the severity of dementia in centenarians. CONCLUSIONS: These results indicate that the neuronal degeneration in very old demented patients involves cortical areas usually preserved at the early stages of the dementing process. Senile plaque formation in certain neocortical areas may be a pathologic hallmark of the severity of dementia in this particular age group. PMID- 7492290 TI - Neuropsychological impairment in Parkinson's disease with and without depression. AB - OBJECTIVES: To compare quantitative and qualitative aspects of neuropsychological test performance in patients with Parkinson's disease who currently had depression (PDD) and those without depression (PDN) so as to evaluate the influence of depression on cognition in Parkinson's disease. DESIGN: Cross sectional comparisons among PDN, PDD, and normal control (NC) groups. The setting was a neurodegenerative disease research center in a teaching hospital. Groups consisted of 44 patients with PDN and 44 patients with PDD matched for age, education, gender, age at onset of disease, disease duration, and disease severity; a group of 44 NC subjects matched for age, education, and gender; and a second set of comparisons between 15 patients with PDN and 15 patients with PDD also matched for overall severity of cognitive impairment. MEASURES: The neuropsychological measures used were the Mattis Dementia Rating Scale, Beck Depression Inventory, Wisconsin Card Sorting Test, Controlled Oral Word Association Test, Logical Memory subtest of the Wechsler Memory Scale-Revised, Digit Span subtest of the Wechsler Adult Intelligence Scale-Revised, and the Boston Diagnostic Aphasia Examination's Animal Naming test and Boston Naming Test. RESULTS: Relative to the NC group, both PDN and PDD groups demonstrated impairments in immediate and delayed verbal recall, semantic fluency, and problem solving. When PDN and PDD groups were matched for demographic and disease variables, only the PDD group evidenced impairment relative to NC in visual confrontation naming, and in lexical and semantic fluency. In addition, impairments on immediate recall and semantic fluency in the PDD group were more pronounced than those in the PDN group. However, when PDN and PDD groups were also matched for overall severity of cognitive impairment, no significant differences emerged among the two groups' neuropsychological test performances. CONCLUSIONS: Depression exacerbates some memory and language impairments associated with PD, even when the PDN and PDD groups are matched for demographic and disease variables. However, the extent and pattern of cognitive impairment is similar in PDN and PDD when the groups are also matched also for overall severity of cognitive impairment. Depression influences the quantity rather than the quality of cognitive impairment associated with Parkinson's disease. PMID- 7492289 TI - Absence of mutations in superoxide dismutase and catalase genes in patients with Parkinson's disease. AB - BACKGROUND: Parkinson's disease (PD) is an adult-onset, neurodegenerative disorder characterized by a selective loss of the dopaminergic cells of the substantia nigra and by progressive motor decline. Studies have shown aberrant oxidative stress metabolism within the substantia nigra and other dopaminergic regions of the brain in patients with PD. OBJECTIVE: To screen the genes of three free radical detoxifying enzymes--copper/zinc superoxide dismutase, manganese superoxide dismutase, and catalase--for mutations in patients with PD. PATIENTS AND METHODS: A total of 107 unrelated patients with PD from two PD populations (familial and sporadic) were screened for mutations in the genes of copper/zinc superoxide dismutase, manganese superoxide dismutase, and catalase by single strand conformation analysis. The diagnosis of PD was based on the clinical observations of resting tremor, rigidity, and bradykinesia. RESULTS: No mutations were identified. However, we did identify an amino acid substitution (glycine to aspartic acid) in exon 9 of the catalase gene in one patient; decreased red blood cell catalase activity was observed in this patient. CONCLUSION: Parkinson's disease is not caused by mutations in the genes of these three detoxifying enzymes. The exon 9 variant in the catalase gene in the one family with PD is most likely a silent mutation and not the genetic cause of PD in this family. PMID- 7492292 TI - Risk of human immunodeficiency virus type 1-related neurologic disease in a cohort of intravenous drug users. AB - BACKGROUND: Although the proportion of cases of acquired immunodeficiency syndrome related to intravenous drug use has increased dramatically over the past decade, there has been no longitudinal examination of primary neurologic disease in this group. OBJECTIVE: To study the development of neurologic disease in human immunodeficiency virus (HIV)-negative and HIV-positive men and women who were intravenous drug users over a 3.5-year period. DESIGN: Prospective observational cohort study. SETTING: Subjects were recruited from an infectious disease clinic at a New York City Hospital or from a methadone maintenance program. PARTICIPANTS: Ninety-nine HIV-negative (62 men and 37 women) and 124 HIV-positive (85 men and 39 women) intravenous drug users volunteered. MAIN OUTCOME MEASURE: The development of clinically significant manifestations in six neurologic domains. RESULTS: With multivariate adjustment for current and past substance abuse, age, education, and head injury, we examined the odds of developing HIV related neurologic disease. Extrapyramidal signs and reduced motor ability became increasingly apparent over time in HIV-infected men as their CD4 cell count declined and as the subjects developed the acquired immunodeficiency syndrome. Fewer neurologic signs were seen in the women. CONCLUSIONS: The impact of HIV infection among intravenous drug users parallels that in homosexual men and is independent of alcohol and other drug use. PMID- 7492291 TI - A comparison of cerebral SPECT abnormalities in HIV-positive homosexual men with and without cognitive impairment. AB - OBJECTIVE: To determine whether technetium Tc 99m exametazime (HMPAO) single photon emission computed tomography (SPECT) can distinguish between human immunodeficiency virus (HIV)-positive homosexual men with normal neuropsychologic test results and HIV-positive homosexual men with abnormal neuropsychologic test results. DESIGN: Neurologic, neuropsychologic, magnetic resonance imaging, and Tc 99m HMPAO SPECT examinations were performed on 10 HIV-positive homosexual men without cognitive impairment and five HIV-positive homosexual men with cognitive impairment. PATIENTS: Human immunodeficiency virus-positive homosexual men from New York City were recruited for the study. MAIN OUTCOME MEASURES: Findings on SPECT scans were evaluated qualitatively for focal defects, heterogeneity of the cortical margin, white matter hypoperfusion, and decreased global cortical uptake. All SPECT focal defects were coregistered with magnetic resonance images; SPECT heterogeneity and global cortical uptake were also measured quantitatively. RESULTS: Coregistration with magnetic resonance imaging revealed that 63% of the focal SPECT defects corresponded to brain gyri and 37% corresponded to sulci. There was no significant difference in the frequency of qualitative or quantitative SPECT abnormalities between HIV-positive homosexual men ith and without cognitive impairment. However, after examining individual neuropsychologic test factors, impaired motor speed performance was associated with decreased quantitative global cerebral uptake. CONCLUSIONS: Qualitative SPECT abnormalities are not increased in frequency in HIV-positive homosexual men with global cognitive impairment compared with those in HIV-positive homosexual men without cognitive impairment. Impaired motor speed performance may be associated with decreased quantitative global cerebral uptake. PMID- 7492293 TI - Complementary positron emission tomographic studies of the striatal dopaminergic system in Parkinson's disease. AB - OBJECTIVE: To assess the relationship between striatal dopa decarboxylase capacity, D2 dopamine receptor binding, and energy metabolism in Parkinson's disease (PD). DESIGN: Positron emission tomographic (PET) studies of glucose and dopa metabolism and D2 dopamine receptor binding in the caudate nucleus and putamen of patients with PD at different Hoehn and Yahr (HY) stages using PET and the tracers 18F-fluorodeoxyglucose (FDG), 6-18F-fluoro-L-dopa (FDOPA), and 11C raclopride (RACLO). SETTING: Positron emission tomography research program at the Paul Scherrer Institute. SUBJECTS: Twenty patients with PD at different stages of the disease (HY stages I through IV; five patients for each stage) compared with separate groups of age-matched healthy subjects. MAIN OUTCOME MEASURES: Influx constant (Ki) for specific FDOPA uptake; uptake index ratio for RACLO binding to D2 dopamine receptors; normalized to global FDG metabolic rate for glucose consumption; and semiquantitative score for assessment of tremor, rigidity, and bradykinesia in PD. RESULTS: Patients with PD at HY stages I to II (hereafter HY I-II PD) revealed reduced FDOPA metabolism, particularly in the putamen. The FDOPA uptake in the putamen and caudate nucleus declined with increasing HY staging and scoring for bradykinesia and rigidity. Putamen RACLO binding to D2 dopamine receptors was up-regulated in patients with HY-I-II PD but declined toward control values, with increasing disease severity. Putamen side-to-side asymmetries of FDOPA metabolism and RACLO binding revealed a significant correlation. Putamen FDG metabolism showed a relative increase in all patients with PD. CONCLUSIONS: Our results show that FDOPA, RACLO, and FDG PET measurements provide complementary information to characterize metabolic and receptor changes in the striatum of PD with different degrees of motor disability. The FDOPA uptake reflects the best motor-related pathologic features, as indicated by the significant correlation between Ki values and clinical scores. The significant association between RACLO and FDOPA in the putamen suggests that D2 dopamine receptor changes are related to the reduction of presynaptic dopaminergic nerve terminals. Putamen FDG increase is probably the result of more complex feedback mechanisms that are primarily induced by striatal dopamine deficiency. PMID- 7492294 TI - Risk factors for impaired outcome after spontaneous intracerebral hemorrhage. AB - OBJECTIVE: To search out independent prognostic factors, including preictal variables for outcome of spontaneous intracerebral hemorrhage. DESIGN: Prospective follow-up study. PATIENTS: One hundred fifty-six consecutive patients (96 men and 60 women) aged 16 to 60 years admitted as emergencies after bleeding. MAIN OUTCOME MEASURES: Potential risk factors (baseline characteristics, health habits, and clinical variables) for death and impaired outcome were studied prospectively up to 1 year after hemorrhage. RESULTS: One year after hemorrhage, 64 patients (41%) were independent and 34 patients (22%) were dependent in the activities of daily living; 58 patients (37%) had died. Risk of death was predicted, after adjustment for sex, age, hypertension, and body mass index, by clinical condition at admission according to the Glasgow Coma Scale (P < .001) and the occurrence of subcortical hematoma (odds ratio [OR], 0.18; 95% confidence interval [CI], 0.04 to 0.91; P = .04). Risk of poor outcome (dependent state or death) was predicted, after adjustment for sex, hypertension, body mass index, cigarette smoking, presence of intraventricular hemorrhage, and surgery, significantly by the Glasgow Coma Scale (P < .001); presence of subcortical hematoma (OR, 0.04; 95% CI, 0.01 to 0.27; P < .001); volume of hematoma (P = .03); age (P = .004); amount of alcohol consumed within 1 week before hemorrhage (P = .03); and presence of cerebellar hematoma (OR, 0.13; 95% CI, 0.02 to 0.95; P = .04). Significant independent predictors of impaired outcome (assessed with the Glasgow Outcome Scale) were the Glasgow Coma Scale (P < .001); presence of subcortical hematoma (OR, 0.26; 95% CI, 0.10 to 0.67; P = .006); alcohol intake within 1 week (P = .002); and presence of cerebellar (OR, 0.16; 95% CI, 0.04 to 0.60; P = .008), intraventricular (OR, 2.74; 95% CI, 1.19 to 6.28; P = .02), or caudate hemorrhage (OR, 0.13; 95% CI, 0.02 to 0.77; P = .03). The mean erythrocyte corpuscular volume was directly associated with an impaired outcome (P < .05). CONCLUSION: In addition to severity and location of the hemorrhage, the age of the patient and the amount of alcohol consumed within 1 week seem to be independent determinants of outcome after intracerebral hemorrhage. PMID- 7492295 TI - Differences in the prevalence of essential tremor among elderly African Americans, whites, and Hispanics in northern Manhattan, NY. AB - BACKGROUND: Until now there has been only one community-based study to examine interethnic differences in the prevalence of essential tremor (ET). The study suggested a higher prevalence among whites than African Americans. The present study is the first to examine differences in the prevalence of ET among Hispanics, African Americans, and whites. OBJECTIVE: To estimate the prevalence of essential tremor (ET) in a cohort of community-dwelling elderly of mixed ethnic background. METHODS: A random sample of 2117 Medicare recipients residing in Washington Heights-Inwood in northern Manhattan, NY, were interviewed. A standardized neurological assessment was performed on those who had neurological complaints and on a random sample of those who did not. Essential tremor was defined as a postural or kinetic tremor of the head or limbs. Diagnoses were independently confirmed by two neurologists based on videotaped examination. RESULTS: After age adjustment to the 1990 Washington Heights-Inwood census, the prevalence of ET was 40.2 per 1000 (95% confidence interval, 31.8 to 48.6). Among 46 cases with ET, ET was significantly more prevalent in men than in women (chi 2 = 5.0, P = .03). Prevalence increased significantly with age. The prevalence was higher in whites than African Americans. The prevalence in Hispanics was intermediate. CONCLUSION: The prevalence of ET increases with age and may be higher among men and whites. Prospective studies are needed to further examine these associations. PMID- 7492296 TI - Myosin heavy-chain fragments and cardiac troponins in the serum in rhabdomyolysis. Diagnostic specificity of new biochemical markers. AB - BACKGROUND: Myosin is the major structural protein in muscle. Antibodies to beta type heavy meromyosin react with cardiac and slow-twitch skeletal muscle. Cardiac TnT and TnI were developed as tissue-specific indicators. OBJECTIVES: To study myosin heavy-chain fragments as a delayed marker of previous rhabdomyolysis. To examine the cardiac specificity of cardiac troponin T (TnT) and cardiac troponin I (TnI) in patients with severe skeletal muscle damage. DESIGN AND METHODS: Serum myosin heavy-chain fragments, TnT, and TnI were studied up to 12 days after diagnosis in relationship to the serum creatine kinase level in 20 patients with rhabdomyolysis. The mean peak serum creatine kinase activity was 91,300 U/L. Myosin heavy-chain fragments were measured by an immunoradiometric assay, TnT by a one-step immunoenzymometric assay, and TnI by an immunoenzymometric assay. RESULTS: Values for serum myosin heavy-chain fragments were greater than the upper limit of normal in all patients. The peak value (70 times the upper normal limit, on average) was usually achieved 4 to 7 days after the diagnosis of rhabdomyolysis, and it was increased up to 12 days. The peak level of TnT was increased in 95% of the patients, and it correlated strongly with the peak activity of serum creatine kinase. The highest TnI value was above the detection limit of myocardial infarction in 30% of the patients. Half of these patients were the only patients with ischemic changes observed on an electrocardiogram performed on admission to the hospital. CONCLUSIONS: The measurement of myosin heavy-chain fragments was useful in the diagnosis of previous rhabdomyolysis up to 12 days. The role of TnT was negligible as an indicator of cardiac muscle damage in patients with severe rhabdomyolysis. Cardiac TnI is a more tissue specific marker for myocardial damage even with concurrent rhabdomyolysis. PMID- 7492297 TI - Airline policy relating to passengers with epilepsy. AB - OBJECTIVE: To identify the policy of international airlines for the carriage of passengers with epilepsy. DESIGN: Postal questionnaire asking about policy for epileptic passengers, training of cabin crew, onboard drugs suitable for the treatment of seizures, and details of any problems reported by crews as a result of in-flight passenger seizures. PARTICIPANTS: The questionnaire was addressed to the medical advisors of 42 international airlines. RESULTS: Thirty (71%) of 42 airlines responded. Eleven (37%) of the 30 airlines that responded had a stated policy or restriction on the carriage of passengers with epilepsy. Five of these airlines advised increasing the dose of anticonvulsant drugs before travel. One airline insisted that epileptic passengers travel with a companion. Twenty-five (83%) of 30 airlines dealt specifically with epilepsy in the training program of their cabin crews. Seventeen (57%) of 30 airlines carried diazepam onboard, mostly in injectable form. Most airlines reported no, or very few, incidents of in-flight passenger seizures annually. CONCLUSIONS: The advice offered to intending epileptic passengers differs greatly between airlines. Some airlines instruct intending passengers to increase their regular medication, probably without justification, since in-flight seizures are very infrequent. Training of cabin crew and the medical equipment carried onboard also vary and in some cases are inadequate. PMID- 7492300 TI - 2',3'-Dideoxycytidine metabolism in a new drug-resistant cell line. AB - 2',3'-Dideoxycytidine (ddC) is a nucleoside analogue that inhibits human immunodeficiency virus type 1 (HIV-1) replication in vitro and is currently used in the therapy of acquired immune deficiency syndrome (AIDS). This compound exerts a delayed cytotoxicity due to inhibition of mitochondrial DNA (mDNA) synthesis. Long-term exposure of U937 human monoblastoid cells to ddC resulted in a time- and concentration-dependent decrease in mDNA content and Rhodamine 123 fluorescence. However, after 2 months on 0.1 microM ddC, a drug-resistant cell line (U937-R) with 66% of the normal amount of mDNA was isolated. ddC transport in U937 and U937-R cell lines was similar. In contrast, U937-R accumulated ddC phosphorylated derivatives at a much lower rate and to a reduced concentration into acid-soluble material. The rate of 2',3'-dideoxycytidine 5'-triphosphate (ddCTP) formation in U937-R cells was almost one-third of that measured in normal cells, although the rate of ddCTP catabolism was similar in both cell lines. Dideoxyliponucleotide (ddCDP-choline and ddCDP-ethanolamine) formation was also much slower (between one-half and one-third as fast) in U937-R than in control cells, although catabolism occurred at similar rates. ddC was phosphorylated by a cytoplasmic deoxycytidine kinase in both cell lines. This enzyme showed Km values for ddC of 80 +/- 7 and 140 +/- 9 microM in U937 and U937-R cells respectively. Furthermore, Vmax was 12 +/- 1.1 and 7.8 +/- 0.5 pmol/min per mg of protein in U937 and U937-R. Thus resistance to ddC toxicity may be due to cells' decreased ability to accumulate intracellular ddC anabolites, which may depend on cytoplasmic deoxycytidine kinase. PMID- 7492299 TI - Glycosylated and unglycosylated human lactoferrins both bind iron and show identical affinities towards human lysozyme and bacterial lipopolysaccharide, but differ in their susceptibilities towards tryptic proteolysis. AB - We studied the role of N-glycosylation of human lactoferrin (hLF) with respect to properties that are relevant to its antibacterial and anti-inflammatory activities. A human kidney-derived 293(S) cell line that constitutively expresses recombinant hLF (rhLF) was produced. The reactivity towards various antibodies of rhLF that had been expressed in the absence or presence of tunicamycin (which blocks N-linked glycosylation) did not differ from that of natural (human milk derived) hLF. Cation-exchange chromatography and N-terminal protein sequencing showed identical cationic properties and an intact N-terminal sequence for rhLF and natural hLF. SDS/PAGE of rhLF expressed in the presence of tunicamycin revealed a protein with the same M(r) as that of enzymically deglycosylated natural hLF. Both glycosylated and unglycosylated rhLF appeared to be completely saturated with iron. The affinity of natural hLF, glycosylated and non glycosylated rhLF for both human lysozyme (Kd 4.5 x 10(-8) M) and bacterial lipopolysaccharide did not differ. SDS/PAGE of hLF species subjected to trypsin indicated that unglycosylated rhLF was much more susceptible to degradation. Furthermore, this analysis suggests that N-glycosylation heterogeneity in natural hLF and rhLF resides in the C-lobe. Thus our results provide no argument for differential antibacterial and/or anti-inflammatory activity of natural and (glycosylated) rhLF and suggest that a major function of glycosylation in hLF is to protect it against proteolysis. PMID- 7492298 TI - Capacitative calcium entry. PMID- 7492301 TI - Regulated and unregulated pathways for MUC2 mucin secretion in human colonic LS180 adenocarcinoma cells are distinct. AB - We have shown previously [McCool, Forstner and Forstner (1994) Biochem. J. 302, 111-118] using pulse-chase labelling of mucin with [3H]threonine that LS180 colonic tumour cells synthesize and secrete MUC2 without the addition of secretagogues. Treatment of the LS180 cells with monensin to disrupt Golgi function was also found to inhibit baseline secretion almost completely. In this paper we show that addition of nocodazole to inhibit microtubule assembly reduced baseline secretion by 53% over a 6 h chase period. In contrast, cytochalasin D did not affect the rate of unstimulated mucin synthesis or secretion, suggesting that baseline secretion is not influenced by disruption of actin microfilaments. In addition, regulated mucin secretion by LS180 cells was studied in response to carbachol, phorbol 12-myristate 13-acetate and A23187. Mucin released in response to secretagogues behaved identically on SDS/PAGE to that secreted under baseline conditions. T84 cells and the B6 subclone of the HT29 cell line responded in a similar manner to LS180 cells and secreted high-molecular-mass mucin which included MUC2 and behaved like LS180 mucin on SDS/PAGE. Neither monensin nor nocodazole significantly affected secretagogue-stimulated mucin secretion. Since these compounds inhibited secretion of labelled mucin under baseline conditions, mucin released by secretagogues must have come from a separate, unlabelled mucin pool in stored granules. Cytochalasin D, on the other hand, caused the release of small amounts of stored mucin, suggesting that actin microfilaments participate in regulated exocytosis. Thus two kinds of mucin secretion occur in LS180 cells. Unregulated secretion depends upon continuous transport of mucin granules from Golgi vesicles to the cell surface and does not utilize stored mucin, whereas regulated secretion involves the release of mucin from storage granules and is not affected by microtubule or Golgi disruption. PMID- 7492302 TI - Cloning and expression of two ornithine decarboxylase forms from HMOA cells. AB - In HMOA cells [Mamont, Duchesne, Grove and Tardif (1978) Exp. Cell Res. 115, 387 393] the half-life of ornithine decarboxylase (ODC) is 8-14 h instead of 15 min as in the Hepatoma Tissue Culture parental cells, due to a single amino acid substitution [Miyazaki, Matsufuji, Murakami and Hayashi (1993) Eur. J. Biochem. 214, 837-844]. We demonstrate for the first time that HMOA cells possess two forms of ODC mRNA that are translated into two proteins differing greatly in turnover rates. We have cloned and transfected the cDNAs for the two ODC forms into COS-1 cells for a direct measurement of their turnover rate. The variant ODC form was much more stable than the wild-type protein, with a half-life of 14 h as compared with 2.5 h. PMID- 7492303 TI - Hyperosmolarity stimulates prostaglandin synthesis and cyclooxygenase-2 expression in activated rat liver macrophages. AB - The effect of aniso-osmotic exposure on the level of inducible cyclooxygenase (Cox-2) and on prostanoid synthesis was studied in cultured rat liver macrophages (Kupffer cells). In lipopolysaccharide (LPS)- or phorbol 12-myristate 13-acetate stimulated Kupffer cells, hyperosmotic (355 mosmol/l) exposure, due to addition of NaCl or impermeant sugars, markedly increased prostaglandin (PG) E2, D2 and thromboxane B2 synthesis in a time- and osmolarity-dependent manner. Increased prostanoid production was observed about 8 h after exposure to LPS in hyperosmotic medium compared to Kupffer cells treated with LPS under normotonic (305 mosmol/l) conditions. A similar stimulatory effect of hyperosmolarity on PGE2 production was also seen when arachidonate was added exogenously. Hyperosmotic stimulation of PGE2 production was accompanied by a strong induction of Cox-2 mRNA levels and an increase in immunoreactive Cox-2, whereas the levels of immunoreactive phospholipase A2 and cyclooxygenase-1 did not change significantly. Dexamethasone, indomethacin and the selective Cox-2 inhibitor, NS 398, abolished the hypertonicity-induced stimulation of PGE2 formation; dexamethasone also prevented the increase in Cox-2 mRNA and protein. The increase of immunoreactive Cox-2 lasted for about 24 h and was also blocked by actinomycin D or cycloheximide, but not by brefeldin A. Tunicamycin or treatment with endoglucosidase H reduced the molecular mass of hypertonicity-induced Cox-2 by 5 kDa. Tunicamycin treatment also suppressed the hypertonicity-induced stimulation of PGE2 production. The hyperosmolarity/LPS-induced stimulation of prostaglandin formation was partly sensitive to protein kinase C inhibition but was not accompanied by an increase in the cytosolic free Ca2+ concentration. The data suggest that osmolarity may be a critical factor in the regulation of Cox-2 expression and prostanoid production in activated rat liver macrophages. PMID- 7492304 TI - Biochemical and pharmacological studies with KT7692 and LY294002 on the role of phosphatidylinositol 3-kinase in Fc epsilon RI-mediated signal transduction. AB - Wortmannin inhibited phosphatidylinositol 3-kinase (P13-kinase) and Fc epsilon RI mediated histamine secretion in RBL-2H3 cells to a similar degree, with IC50 values of 3 and 2 nM, respectively. Although P13-kinase is an acknowledged regulator of intracellular trafficking and secretion, wortmannin has proved to be a difficult drug to use in assessing the role of P13-kinase because it inhibits another important enzyme, myosin light-chain kinase (MLCK; IC50 = 200 nM). In the present study we synthesized a unique derivative of wortmannin, O-acetyl-delta 16 wortmannin-17-ol (KT7692), that has an inhibitory potency against PI3-kinase one hundredth that of wortmannin, but retains a similar potency to wortmannin against MLCK. Histamine secretion was influenced 100-fold more by wortmannin than by KT7692.2-(4-Morpholinyl)-8-phenyl-4H-1-benzopyran-4-one (LY294002), a structurally different PI3-kinase inhibitor from wortmannin, inhibited PI3-kinase with an IC50 of 2 microM but had little effect on MLCK activity in this concentration range. LY294002 also inhibited histamine secretion in RBL-2H3 cells with an IC50 of 5 microM. These results provide further evidence that PI3-kinase is involved in the signal transduction pathway responsible for histamine secretion after stimulation of Fc epsilon RI. Furthermore KT7692 in combination with wortmannin and LY294002 would be a powerful tool for clarifying the involvement of PI3-kinase as distinct from that of MLCK in signal transduction systems of various cellular responses. PMID- 7492305 TI - Ca2+ signalling in K562 human erythroleukaemia cells: effect of dimethyl sulphoxide and role of G-proteins in thrombin- and thromboxane A2-activated pathways. AB - The human leukaemic cell line K562 is a pluripotent stem cell with the potential to mature along a megakaryocytic or erythroid line. In these cells, thrombin and U46619 (9,11-dideoxy-9 alpha, 11 alpha-methanoepoxy prostaglandin F2 alpha), a thromboxane A2 analogue, increased intracellular Ca2+ in a rapid and concentration-dependent manner. The peak transient observed with both thrombin and U46619 was preserved upon stimulation in the absence of extracellular calcium and blunted with phorbol myristate acetate, suggestive of activation of phospholipase C. Short-term treatment with leupeptin abolished the calcium response to thrombin, but did not alter that to U46619. Both pertussis toxin (PT) and DMSO pretreatment inhibited thrombin- but not U46619-stimulated intracellular calcium elevation, indicating that these agonists signal through different G proteins. Western blot analysis of crude membranes from K562 cells revealed the presence of G12 alpha and G13 alpha; the other known PT-substrates, Gi1 alpha and G0 alpha, were not detected. Consistent with this observation, ADP-ribosylation experiments revealed the presence of two PT substrates which co-migrated with human erythrocyte G12 alpha and G13 alpha. An antibody raised against Gq/11 alpha, a subfamily of G-protein alpha subunits unmodified by PT, specifically recognized 42 kDa protein(s) in K562 cells. PCR amplification of reverse transcribed K562 RNA followed by DNA sequencing showed that these cells express messages for both Gq alpha and G11 alpha. Treatment of K562 cells with DMSO reduced the levels of thrombin receptor mRNA, without simultaneous changes in the expression of G12 alpha and G13 alpha. We have thus identified Ca(2+)-mobilizing agonists and related G-proteins in K562 cells, together with changes induced by DMSO in this signalling pathway. PMID- 7492306 TI - Role of phosphatidylinositol 3-OH-kinase activity in the inhibition of apoptosis in haemopoietic cells: phosphatidylinositol 3-OH-kinase inhibitors reveal a difference in signalling between interleukin-3 and granulocyte-macrophage colony stimulating factor. AB - Apoptosis of haemopoietic cells can be inhibited by various cytokines, but the specific signalling pathways involved are not well defined. Interleukin (IL)-4 has unique actions since it is unable to activate p21ras or mitogen-activated protein kinase, but can activate PtdIns 3-OH-kinase (PtdIns 3-kinase), the latter effect being shared with most other cytokines. In many cases, IL-4 is able to maintain cell viability by inhibiting apoptosis, but is unable to stimulate continuous proliferation of cells. This led us to investigate the role of PtdIns 3-kinase in inhibition of apoptosis. Two potent inhibitors of PtdIns 3-kinase, wortmannin and LY294002, rapidly induced apoptosis in cells incubated in the presence of IL-4, at concentrations consistent with their ability to inhibit PtdIns 3-kinase activity in whole cells. Interestingly, the same effect was seen in cells in the presence of IL-3 and Steel factor, while cells incubated in the presence of granulocyte-macrophage colony stimulating factor, and to a lesser extent, IL-5 could bypass the effect of wortmannin or LY294002. Therefore, this study suggests that PtdIns 3-kinase activity provides an important, although not a unique signal, required to inhibit apoptosis in haemopoietic cells. PMID- 7492307 TI - A hierarchy of ATP-consuming processes in mammalian cells. AB - The rates of different ATP-consuming reactions were measured in concanavalin A stimulated thymocytes, a model system in which more than 80% of the ATP consumption can be accounted for. There was a clear hierarchy of the responses of different energy-consuming reactions to changes in energy supply: pathways of macromolecule biosynthesis (protein synthesis and RNA/DNA synthesis) were most sensitive to energy supply, followed by sodium cycling and then calcium cycling across the plasma membrane. Mitochondrial proton leak was the least sensitive to energy supply. Control analysis was used to quantify the relative control over ATP production exerted by the individual groups of ATP-consuming reactions. Control was widely shared; no block of reactions had more than one-third of the control. A fuller control analysis showed that there appeared to be a hierarchy of control over the flux through ATP: protein synthesis > RNA/DNA synthesis and substrate oxidation > Na+ cycling and Ca2+ cycling > other ATP consumers and mitochondrial proton leak. Control analysis also indicated that there was significant control over the rates of individual ATP consumers by energy supply. Each ATP consumer had strong control over its own rate but very little control over the rates of the other ATP consumers. PMID- 7492308 TI - Inhibition of cytochrome c oxidase in turnover by nitric oxide: mechanism and implications for control of respiration. AB - Binding of nitric oxide (NO) to isolated cytochrome c oxidase in turnover was investigated by static and kinetic spectroscopic methods. These studies indicate that cytochrome c oxidase rapidly binds NO when the enzyme enters turnover. Our results show that NO binds to ferrocytochrome a3, competing with oxygen for this binding site. However, the main features of the binding process, in particular the rapid onset of inhibition, cannot be fully explained on this basis. We suggest, therefore, that there is a second binding site for NO, which has lower affinity but nevertheless plays an important role in the inhibitory process. A likely possibility is that CuB+ constitutes this second binding site. The fast onset of inhibition observed in the presence of NO, along with the dependence on the oxygen concentration, suggests that under physiological conditions, where the oxygen concentration is low, nanomolar concentrations of NO can effectively act as a regulator of the mitochondrial respiratory chain. PMID- 7492309 TI - Expression cloning of a zinc-finger cyclic AMP-response-element-binding protein. AB - In response to specific extracellular signals, intracellular cyclic AMP levels increase, leading to a variety of responses including the alteration of transcription of many eukaryotic genes. This transcriptional effect is frequently mediated through the cyclic AMP-response element (CRE) motif T(T/G)ACGTCA. Using an expression screening approach we have cloned a yeast gene, MSN2, that encodes a 78 kDa protein that recognizes this consensus CRE motif. Phosphorylation of the MSN2 protein by the catalytic subunit of protein kinase A stimulates DNA binding in vitro. Two putative Cys2His2-type zinc fingers present in the C-terminal 79 amino acids of the MSN2 protein are sufficient to confer CRE-binding specificity. Therefore, MSN2 represents a novel CRE-binding protein distinct from the multiple previously characterized basic region-leucine zipper repeat CRE-binding proteins. PMID- 7492310 TI - Ca(2+)-dependent binding of annexin IV to surfactant protein A and lamellar bodies in alveolar type II cells. AB - Surfactant protein A (SP-A), a lung-specific glycoprotein in pulmonary surfactant, is synthesized and secreted from the alveolar type II cells. It has been shown that SP-A is a Ca(2+)-binding protein with several binding sites and that the high-affinity site(s) is located in the C-terminal region of SP-A. In the present study we isolated the proteins from bovine lung soluble fraction that bind to SP-A in a Ca(2+)-dependent manner using DEAE-Sephacel and SP-A-conjugated Sepharose 4B. At least three different protein bands with molecular masses of 24.5, 32, and 33 kDa were observed on SDS/PAGE. The main protein, with molecular mass of 32 kDa, was identified as annexin IV by the partial-amino-acid-sequence analyses and an immunoblot analysis with anti-(annexin IV) antiserum. We also found from the immunoblot analysis that the cytosolic fraction of isolated rat alveolar type II cells contains annexin IV. In addition, when rat lung cytosol was loaded on to the lung lamellar body-conjugated Sepharose 4B in the presence of Ca2+, two proteins, with molecular masses of 32 and 60 kDa on SDS/PAGE respectively, were eluted with EGTA. The 32 kDa protein was shown to be annexin IV by an immunoblot analysis with the antiserum against annexin IV. The lung annexin IV augmented the Ca(2+)-induced aggregation of the lung lamellar bodies from rats. However, the augmentation of aggregation of the lung lamellar bodies by annexin IV was attenuated when the lamellar bodies were preincubated with polyclonal anti-SP-A antibodies. SP-A bound to annexin IV under conditions where contaminated lipid was removed. These results suggest that SP-A bound to annexin IV based on protein-protein interaction, though both proteins are phospholipid binding proteins. All these findings suggest that the interaction between SP-A and annexin IV may have some role in alveolar type II cells. PMID- 7492312 TI - Design of a new protease inhibitor by the manipulation of the bait region of alpha 2-macroglobulin: inhibition of the tobacco etch virus protease by mutant alpha 2-macroglobulin. AB - Human alpha 2-macroglobulin (alpha 2M) inhibits a broad spectrum of proteases by changing its conformation and physically confining the enzyme. The inhibitory spectrum of alpha 2M is defined by a stretch of 39 amino acids, the bait region, located near the middle of the alpha 2M monomers. To investigate whether a new inhibitory specificity can be introduced by the manipulation of the bait region, recombinant alpha 2M (r alpha 2M) was produced in which the primary cleavage site was replaced by a heptapeptide containing the cleavage specificity of tobacco etch virus (TEV) protease. This protease is not inhibited by wild-type alpha 2M. The r alpha 2M, produced in an episomal expression system, was fully functional and able to inhibit the tobacco etch virus protease according to its normal 'trap' mechanism. The manipulation of the bait region of alpha 2M thus allows the design of new, specific protease inhibitors. PMID- 7492313 TI - The activity of calmodulin is altered by phosphorylation: modulation of calmodulin function by the site of phosphate incorporation. AB - Calmodulin transduces Ca2+ signals by binding to and activating essential regulatory enzymes. The large number of intracellular targets for calmodulin raises the possibility that mechanisms in addition to Ca2+ may modulate calmodulin activity. Phosphocalmodulin is found in cells and tissues, and calmodulin phosphorylation is enhanced by several mitogens. Phosphorylation of calmodulin on serine/threonine residues by casein kinase II decreased its ability to activate Ca2+/calmodulin-dependent protein kinase II (CaM-kinase II). The major effect was a 2.5-fold increase in the concentration at which half-maximal velocity (K0.5) was attained, with no apparent alteration in the Vmax, or the K0.5 for Ca2+. In contrast, calmodulin phosphorylated on tyrosine residues by the insulin receptor kinase produced an increase in the Vmax, with no alteration in the affinity for CaM-kinase II or the K0.5 for Ca2+. Direct determination by surface plasmon resonance of the dissociation constants with a synthetic peptide corresponding to the calmodulin-binding domain of CaM-kinase II revealed that phosphorylation on serine/threonine residues of calmodulin significantly decreased its affinity for the peptide, while tyrosine phosphorylation had no effect on binding. In contrast to CaM-kinase II, neither serine/threonine nor tyrosine phosphorylation of calmodulin altered its ability to activate calcineurin. These data indicate that phosphorylation of calmodulin differentially modifies its interaction with individual target enzymes. Moreover, the amino acid residues phosphorylated provide an additional level of control. These results demonstrate that phosphorylation is an in vitro regulatory mechanism in the targeting of calmodulin responses and, coupled with the stoichiometric phosphorylation of calmodulin in rat hepatocytes, suggest that it may be relevant in intact cells. PMID- 7492311 TI - Synthesis of a Cys949Tyr alpha 2-macroglobulin thiol ester mutant: co transfection with wild-type alpha 2-macroglobulin in an episomal expression system. AB - A full-length alpha 2-macroglobulin (alpha 2M) cDNA was cloned into the episomal expression vectors pREP7 and pMEP4. Electroporation of the cell lines WI-L2 729HF2, U-937, K-562 and an Epstein-Barr virus-transformed cell line resulted in stable transfectants only with K-562 cells. Stable expression was obtained exclusively with pMEP4-alpha 2M and was driven from the inducible human metallothionein IIA promoter. Expression of the wild-type alpha 2M cDNA resulted in a recombinant protein (r alpha 2M) that could not be distinguished from plasma alpha 2M (p-alpha 2M): the transfected K-562 cells secreted tetrameric alpha 2M with intact internal thiol esters, a functional bait domain and a latent receptor binding domain. r alpha 2M inhibited trypsin and elastase from cleaving a high molecular-mass substrate. When the Cys-949 involved in the formation of the internal thiol ester was mutated to tyrosine (C949Y-r alpha 2M), a tetrameric alpha 2M was secreted, with the electrophoretic mobility of methylamine-treated p alpha 2M (p-alpha 2M/MA) and with a functional receptor-binding domain. The C949Y r alpha 2M did not possess proteinase-inhibiting capacity. Heterozygosity was mimicked by co-transfecting the K-562 cells with wild-type and mutant expression vectors. In this case, r alpha 2M was secreted with zero, one, two, three or four internal thiol esters. A comparison of the interaction of interleukin 1 beta and basic fibroblast growth factor with native p-alpha 2M, p-alpha 2M/MA and the mutant C949Y-r alpha 2M revealed that when assayed under nondenaturing conditions, no binding occurred to 'slow' p-alpha 2M whereas quantitatively similar binding was observed to 'fast' p-alpha 2M/MA and C949Y-r alpha 2M. Covalent binding, however, was essentially limited to p-alpha 2M/MA, suggesting the involvement of Cys-949 in the process. Covalent binding of insulin, on the contrary, was only observed when it was present during hydrolysis of the internal thiol esters of p-alpha 2M by trypsin treatment, and thus involves the activated Glx residue. PMID- 7492314 TI - Translocon-associated protein TRAP delta and a novel TRAP-like protein are coordinately expressed with pro-opiomelanocortin in Xenopus intermediate pituitary. AB - In the intermediate pituitary gland of Xenopus laevis, the expression levels of the prohormone pro-opiomelanocortin (POMC) can be readily manipulated. When the animal is placed on a black background, the gene for POMC is actively transcribed, whereas on a white background the gene is virtually inactive. In this study, we characterized two genes whose transcript levels in the intermediate pituitary are regulated in coordination with that for POMC. One of these codes for a protein homologous to translocon-associated protein TRAP delta, a subunit of a transmembrane protein complex located at the site where nascent secretory proteins enter the endoplasmic reticulum (ER). Both Xenopus and mice were found to express an alternatively spliced transcript that gives rise to a previously unknown version of the TRAP delta protein. The product of the second gene is a novel and highly conserved protein with structural similarity to glycoprotein gp25L, a constituent of another translocon-associated protein complex. A database search revealed the existence of a novel family of gp25L related proteins whose members occur throughout the animal kingdom. Together, our data imply that (i) the group of ER proteins surrounding translocating polypeptide chains may be far more complex than previously expected, and (ii) a number of the accessory components of the translocon participate in early steps of prohormone biosynthesis. PMID- 7492315 TI - Tilorone-induced lysosomal storage of glycosaminoglycans in cultured corneal fibroblasts: biochemical and physicochemical investigations. AB - Tilorone (2,7-bis[2-(diethylamino)ethoxy]-fluoren-9-one) and several other bis basic compounds are known to induce lysosomal glycosaminoglycan (GAG) storage. The responsible pathomechanism has not been elucidated yet. The assumption of an unspecific disturbance of lysosomal proenzyme targeting due to elevation of endosomal pH is opposed by the hypothesis of formation of a complex between tilorone and GAGs within the lysosomes, which renders GAGs indigestible to glycosidases. In cultures of bovine corneal fibroblasts the amounts of intracellular GAGs [dermatan sulphate (DS), heparan sulphate (HS) and chondroitin sulphate (CS)] were quantified. The fibroblasts were exposed to tilorone (5 microM), which was found to be readily taken up by the cells and to be accumulated within acidic compartments to finally achieve millimolar concentrations. Under these conditions the GAG storage is predominantly due to the accumulation of DS; however, the DS secretion into the culture medium was not affected. The HS accumulation was much less pronounced, accounting only for 3% of total GAG storage. Ammonium chloride (10 mM), which is known to diminish lysosomal enzyme activity by interfering with the mannose 6-phosphate receptor mediated transport, prevents both HS and DS breakdown. By means of NMR spectroscopy it was shown that tilorone itself tends to display a concentration dependent aggregation which was enhanced in the presence of GAGs. The diethylamino groups of tilorone interact physicochemically with DS, and to a smaller extent with HS, but not with chondroitin 4-sulphate. Thus, the strength of the interaction between tilorone and the different GAGs in vitro correlates with the potency of tilorone to inhibit the breakdown of the individual GAGs in cultured bovine fibroblasts. The results support the hypothesis of a specific interaction between tilorone and particular GAGs, rendering these resistant to enzymic degradation. PMID- 7492316 TI - The integrin alpha IIb beta 3 contains distinct and interacting binding sites for snake-venom RGD (Arg-Gly-Asp) proteins. Evidence that the receptor-binding characteristics of snake-venom RGD proteins are related to the amino acid environment flanking the sequence RGD. AB - We have previously demonstrated [Lu, Williams, Deadman, Salmon, Kakkar, Wilkinson, Baruch, Authi and Rahman (1994) Biochem. J. 304, 929-936] the preferential antagonism of the interactions of the integrin alpha IIb beta 3 on activated platelets with three immobilized glycoprotein ligands (fibrinogen, fibronectin and von Willebrand factor) by a selected panel of snake-venom RGD (Arg-Gly-Asp)-containing proteins including the disintegrins kistrin and elegantin, and the neurotoxin variant dendroaspin. Kistrin and dendroaspin, although structurally unrelated, contain similar amino acids flanking the tripeptide RGD and behaved as identical antagonists preferentially inhibiting platelet adhesion to immobilized fibrinogen as opposed to fibronectin. In contrast, elegantin, which shares extensive sequence similarity with kistrin but has different amino acids around the tripeptide RGD, preferentially inhibited platelet adhesion to immobilized fibronectin as opposed to fibrinogen. To develop further insights into the mechanisms underlying the preferential antagonism shown by the venom proteins in the adhesion studies, we, in the present study, sought to determine the binding properties of kistrin, elegantin and dendroaspin to the alpha IIb beta 3 complex by radioligand kinetic and competition studies. In direct binding experiments, both kistrin and dendroaspin were observed to bind to a single class of binding site on ADP-activated platelets with apparent equilibrium dissociation constant (Kdapp) values of 42 +/- 2 nM and 21 +/- 6 nM respectively. In competition studies, dendroaspin blocked the binding of 125I labelled kistrin to ADP-activated platelets in a simple competitive manner, with an apparent equilibrium inhibition constant (Kiapp) of 143 +/- 14 nM, from which an indirect Kdapp = 22 nM for dendroaspin was determined. This result suggests that kistrin and dendroaspin bind to the same site on the integrin alpha IIb beta 3 consistent with their similar inhibitory properties. In contrast, elegantin recognized two classes of binding sites on the alpha IIb beta 3 complex with Kdapp values of 10.5 +/- 0.8 nM and 175 +/- 10 nM, and, unlike dendroaspin, did not inhibit the binding of 125I-labelled kistrin to ADP-activated platelets. However, in reciprocal experiments both kistrin and dendroaspin inhibited the binding of 125I-elegantin to ADP-activated platelets in a non-competitive manner, with Kiapp values of 34 +/- 3 nM and 21 +/- 2 nM respectively. Thus elegantin appears to interact with distinct but interacting sites on the alpha IIb beta 3 complex from the binding site of kistrin and dendroaspin, consistent with its distinctive inhibitory preferences as shown in platelet adhesion studies.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 7492318 TI - Evaluation of the extent of the binding site in human tissue kallikrein by synthetic substrates with sequences of human kininogen fragments. AB - We have synthesized internally quenched peptides spanning the Met379-Lys380 or Arg389-Ser390 bonds of human kininogen (hkng) that flank lysyl-bradykinin and have studied the kinetics of their hydrolysis by human tissue kallikrein. The kinetic data for the hydrolysis of the Met-Lys bond in substrates with an N terminal extension showed that interactions up to position residue P10 contribute to the efficiency of cleavage. In contrast, there were no significant variations in the kinetic data for the hydrolysis of substrates with C-terminal extensions at sites P'4 to P'11. A similar pattern was observed for the cleavage of substrates containing an Arg-Ser bond because substrates extended up to residue P6 were hydrolysed with the highest kcat/Km values in the series, whereas those extended to P'11 on the C-terminal side had a lower susceptibility to hydrolysis. Time-course studies of hydrolysis by human and porcine tissue kallikreins of a Leu373 to Ile393 human kininogen fragment containing omicron-aminobenzoic acid (Abz) at the N-terminus and an amidated C-terminal carboxyl group Abz-Leu-Gly-Met Ile-Ser-Leu-Met-Lys-Arg- Pro-Pro-Gly-Phe-Ser-Pro-Phe-Arg-Ser-Ser-Arg-Ile-NH2 (Abz [Leu373-Ile393]-hkng-NH2) indicated that the cleavage of Met-Lys and Arg-Ser bonds in the same molecule occurs via the formation of independent enzyme substrate complexes. The hydrolysis of Abz-F-R-S-S-R-Q-EDDnp [where EDDnp is N (2,4-dinitrophenyl)ethylenediamine] and Abz-M-I-S-L-M-K-R-P-Q-EDDnp by human tissue kallikrein had maximal kcat/Km values at pH 9-9.5 for both substrates. The pH-dependent variations in this kinetic parameter were almost exclusively due to variations in kcat. A significant decrease in kcat/Km values was observed for the hydrolysis of Arg-Ser and Met-Lys bonds in the presence of 0.1 M NaCl. Because this effect was closely related to an increase in Km, it is likely that sodium competes with the positive charges of the substrate side chains for the same enzyme subsites. PMID- 7492317 TI - Characterization of N-ethylmaleimide-sensitive thiol groups required for the GTP dependent fusion of endoplasmic reticulum membranes. AB - The GTP-dependent fusion activity of endoplasmic reticulum membranes is thought to be required for the structural maintenance and post-mitotic regeneration of the endoplasmic reticulum. This fusion is sensitive to the thiol-alkylating agent N-ethylmaleimide. In many intracellular fusion events N-ethylmaleimide sensitivity is associated with a homotrimeric ATPase called N-ethylmaleimide sensitive fusion protein or NSF. The addition of cytosol containing NSF is known to restore fusion activity to N-ethylmaleimide-treated membranes. We found that the inhibition of fusion of rat liver endoplasmic reticulum membranes (microsomes) by N-ethylmaleimide was not reversed by the addition of untreated cytosol. Fusion was also unaffected by treatment with a buffer known to remove NSF from membranes. Accordingly, no membrane-associated NSF was detected by immunoblot analysis. These data suggest that microsome fusion requires an N ethylmaleimide-sensitive component distinct from NSF. This component was tightly associated with the membranes, so we used a number of chemical probes to characterize it in situ. Its thiol groups did not appear to be part of a GTP binding site. They showed relatively low reactivity with sodium periodate, which induces the formation of disulphide bonds between proximate thiol groups. The thiols were not protected against N-ethylmaleimide by Zn2+, a potent inhibitor of fusion which is known to efficiently co-ordinate thiol groups. To characterize the topology of the fusion-related thiol groups we used bulky thiol-specific reagents prepared by conjugating BSA or 10 kDa aminodextran to the bifunctional reagent N-succinimidyl 3-(2-pyridyldithio)propionate. The inhibition of fusion by these reagents indicated that these thiols are highly exposed on the membranes. This exposure might be important for the function of these groups during GTP triggered fusion. PMID- 7492319 TI - Analysis of transcripts encoding novel members of the mammalian metalloprotease like, disintegrin-like, cysteine-rich (MDC) protein family and their expression in reproductive and non-reproductive monkey tissues. AB - A number of sequence-related, cysteine-rich proteins containing metalloprotease like and disintegrin-like domains (the MDC protein family), at least one of which has been shown to play a role in egg recognition during fertilization, are abundantly expressed in the mammalian male reproductive tract. In this paper we report the cloning and sequence analysis of three closely related isoforms of a novel member of this family which are expressed not only in the testis, but also in the liver, albeit at a lower level. Using a PCR-based approach we also demonstrate the presence of transcripts encoding additional, novel, disintegrin containing proteins, in the liver and epididymis. We conclude that while some members of the MDC family are specific to the reproductive tract, suggesting functions peculiar to those tissues, others have a broader tissue distribution and may therefore play a more general role in integrin-mediated cell-cell recognition, adhesion or signalling. PMID- 7492320 TI - Effects of aging on the synthesis of antithrombin-binding sites on heparin chains and heparan sulphate chains in the rat. AB - [35S]Heparin proteoglycans were isolated from the skins and peritoneal mast cells of male rats aged 2 to 22 months. Their [35S]heparin chains were separated on antithrombin-agarose into fractions with high and low affinities for antithrombin. In skin, the proportion of 35S-labelled high-affinity heparin chains declined from 23% at 2 months to 8% at 12 months and did not change significantly between 12 and 22 months. In peritoneal mast cells, the proportion of 35S-labelled high-affinity heparin chains increased from 14% at 2 months to 21% at 4 months and then did not vary significantly until 15 months of age. By 21 months a consistent and significant decline to 8% occurred. The structures of high-affinity heparin proteoglycans did not change with age. Their decreased proportions, without change in their structure, may indicate that they are produced by a unique subset of mast cells, the proportion of which declines with age. [35S]heparan sulphate chains were isolated from skins and brains of rats in the same age range and fractionated on antithrombin-agarose. There were no significant variations in the proportions of 35S-labelled high-affinity heparan sulphate chains in skin (10%) or brain (24%) between 4 and 22 months of age. PMID- 7492323 TI - Effects of ionic strength on the catalysis and stability of prolyl oligopeptidase. AB - Prolyl oligopeptidase is the prototype of a new serine protease family, unrelated to trypsin and subtilisin. In contrast with these proteases, prolyl oligopeptidase is remarkably sensitive to ionic strength, being more active in the presence of high concentrations of salt. The enzyme has two catalytic forms, which interconvert with changing pH. To reveal the structural bases of these phenomena, the effects of 0.5 M NaCl on the stability of the enzyme were investigated by studying its denaturation as a function of pH, temperature, and urea concentration. The three independent methods have unequivocally demonstrated that denaturation of the enzyme is promoted in the presence of NaCl. Furthermore, destabilization of the low-pH form by urea is more significant than that of the high-pH form. Examination of the fluorescence emission spectra of various denatured forms indicates that the enzyme is not fully unfolded in 8 M urea, nor at acidic pH. The tryptophan residues in the acid-denatured state are mainly buried. The results are interpreted in terms of the decay of the protective water shell at the higher ionic strength. The higher enthalpy and entropy of activation for heat denaturation provide further evidence that a more ordered water structure stabilizes the protein in the absence of salt. The biphasic kinetics obtained with denaturation by heat and urea suggest that the enzyme has two domains of different stabilities. PMID- 7492322 TI - The interaction of methanol dehydrogenase and its cytochrome electron acceptor. AB - A fluorescence method is described for direct measurement of the interaction between methanol dehydrogenase (MDH) and its electron acceptor cytochrome cL. This has permitted a distinction to be made between factors affecting electron transfer and those affecting the initial binding or docking process. It was confirmed that the initial interaction is electrostatic, but previous conclusions with respect to the mechanism of EDTA inhibition have been modified. It is proposed that the initial 'docking' of MDH and cytochrome cL is by way of ionic interactions between lysyl residues on its surface and carboxylate groups on the surface of cytochrome cL. This interaction is not inhibited by EDTA, which we suggest acts by binding to nearby lysyl residues, thus preventing movement of the 'docked' cytochrome to its optimal position for electron transfer, which probably involves interaction with the hydrophobic funnel in the surface of MDH. PMID- 7492321 TI - Modulation of CD4 lateral mobility in intact cells by an intracellularly applied antibody. AB - This study shows that the lateral mobility of CD4, an important plasma-membrane immune receptor, can be modulated by intracellular application of an anti-CD4 antibody. For this purpose, (i) full-length CD4 and a truncated CD4 mutant, lacking a 32-residue-long C-terminal intracellularly exposed domain, were expressed in Spodoptera frugiperda (Sf9) insect cells, (ii) a monoclonal antibody, C6, with specificity for the C-terminal domain was generated, and (iii) a versatile apparatus for fluorescence microphotolysis (FM) studies was constructed. By these means it was found that the commercial anti-CD4 antibody Leu3a-PE, in contrast with several other anti-CD4 antibodies, could be used as a fluorescent label of CD4 without interfering greatly with CD4 mobility. Labelled by Leu3a-PE, full-length CD4 had a lateral diffusion coefficient of D = (4.7 +/- 1.9) x 10(-10) cm2/s and a mobile fraction of fm = 80 +/- 16% (room temperature). Within experimental accuracy the truncated CD4 had the same mobility as full length CD4. Introduction of the C6 antibody into Sf9 cells by microinjection or by fusion with C6-loaded liposomes decreased the mobility of full-length CD4 (fm = 40%) but not of truncated CD4 (fm = 80%). Treatment of Sf9 cells with phorbol ester also reduced the mobility of full-length CD4 (fm = 50%) but not truncated CD4 (fm = 90%). A calmodulin inhibitor but not a protein kinase C (PKC) inhibitor abolished the phorbol ester effect. PMID- 7492324 TI - The effects of the site-directed removal of N-glycosylation sites from beta-1,4-N acetylgalactosaminyltransferase on its function. AB - The amino acid sequence deduced from the cloned human cDNA of beta-1,4-N acetylgalactosaminyltransferase (GalNAc-T; EC 2.4.1.92) gene predicted three potential sites for N-linked glycosylation. Although many glycosyltransferases isolated contain from 2 to 6 N-glycosylation sites, their significance has not been adequately demonstrated. To clarify the roles of N-glycosylation in GalNAc-T function, we generated a series of mutant cDNAs, in which some or all of the glycosylation recognition sites were eliminated by polymerase chain reaction (PCR)-mediated site-directed mutagenesis. Using transcription/translation in vitro, we confirmed that all potential N-glycosylation sites could be used. Although cell lines transfected with mutant cDNAs showed equivalent levels of GalNAc beta 1-->4(NeuAc alpha 2-->3)Gal beta 1-->4Glc-Cer (GM2) to that of the wild-type, the extracts from mutant cDNA transfectants demonstrated lower enzyme activity than in the wild-type. The decrease in enzyme activity was more evident as the number of deglycosylated sites increased, with about 90% decrease in a totally deglycosylated mutant. The enzyme kinetics analysis revealed no significant change of Km among wild-type and mutant cDNA products. The intracellular localization of GalNAc-T expressed in transfectants with wild-type or mutant cDNAs also showed a similar perinuclear pattern (Golgi pattern). These results suggest that N-linked carbohydrates on GalNAc-T are required for regulating the stability of the enzyme structure. PMID- 7492325 TI - Catalysis by the large subunit of the second beta-galactosidase of Escherichia coli in the absence of the small subunit. AB - Plasmids containing the ebgAo and ebgAa genes of Escherichia coli under the control of the lac repressor and promoter have been constructed and inserted into Salmonella typhimurium CH3. This system expresses the large subunit of the ebgo and ebga beta-galactosidase in high yield (20-60% of total protein). The large subunits have been purified to homogeneity. As isolated they are tetramers of significant catalytic activity; the N-terminal amino acid residue is Met, but it is not formylated. The kcat. values for a series of aryl galactosides were 6-200 fold reduced from the corresponding values for the holoenzymes. kcat/Km Values for glycosides of acidic aglycones, though, were unchanged, whilst kcat./Km values for galactosides of less acidic aglycones showed a modest (up to 10-fold) decrease. The kcat. values for glycosides of acidic aglycones hydrolysed by ebgo and ebga large subunits were essentially invariant with aglycone pK, suggesting that hydrolysis of the galactosyl-enzyme intermediate had become rate-determining for these substrates. Rate-determining hydrolysis of the glycosyl-enzyme intermediate was confirmed by pre-steady-state measurements and nucleophilic competition with methanol. Absence of the small subunit was thus estimated to cause a 200-fold decrease in degalactosylation rate for ebgo and a 20-fold one for ebga. beta 1g(V/K) values of -0.57 +/- 0.08 for ebgo and -0.54 +/- 0.08 for ebga isolated subunits were significantly more negative than for holoenzymes. It is suggested that the small subunit is associated with the optimal positioning of the electrophilic Mg2+ ions in these enzymes. Use of PCR in the construction of the plasmid also inadvertently led to the production of psi ebgo large subunit in which there was a PCR-introduced Leu9-->His change. Values of kcat. for aryl galactosides, calculated on the assumption that the psi ebgo large subunit, like the ebgo and ebga large subunits, was 100% active as isolated, were about an order of magnitude lower than for true ebgo large subunit, whilst Km values were similar. The very significant kinetic effect of this inadvertant site-undirected mutagenesis indicates that quite large kinetic effects of amino-acid replacements in enzymes may have no obvious mechanistic significance. PMID- 7492326 TI - Glycerol 3-phosphate-induced ATP production in intact mitochondria from pancreatic B-cells. AB - A bioluminescent method is presented that allows monitoring of ATP production from mitochondria corresponding to one islet of Langerhans per sample. In mitochondria from ob/ob mice Ca2+ stimulates the ATP production in the presence of L-glycerol 3-phosphate (GP) by reducing the Km for GP by one order of magnitude to about 3 mM. Maximal ATP production in the presence of Ca2+ (200 nM) is obtained at 10 mM GP. The free calcium concentration required to reach half maximal stimulation (K0.5Ca2+) depends on the GP concentration, thus half-maximal effects are observed at about 80 nM at low GP (1 mM) and 10 nM at high GP (10 mM). Sodium can replace Ca2+ as a stimulator of GP-induced ATP production. It activates ADP phosphorylation by B-cell mitochondria in a sigmoidal concentration dependent manner in the absence of Cs2+ (Hill coefficient 2.3 +/- 0.2) but does not change K0.5ca2+ nor the maximal mitochondrial activity. Ca2+ concentrations higher than 300 nM are inhibitory at all tested substrate concentrations. Mitochondria from ob/ob mice showed no functional defect when compared with normal controls. It is concluded that activation of the glycerol phosphate shuttle may not be the main coupling site for glucose-induced insulin release at maximal cytoplasmic Ca2+ levels. PMID- 7492327 TI - Amino acid sequence and the cellular location of the Na(+)-dependent D-glucose symporters (SGLT1) in the ovine enterocyte and the parotid acinar cell. AB - The Na(+)-dependent D-glucose symporter has been shown to be located on the basolateral domain of the plasma membrane of ovine parotid acinar cells. This is in contrast to the apical location of this transporter in the ovine enterocyte. The amino acid sequences of these two proteins have been determined. They are identical. The results indicated that the signals responsible for the differential targeting of these two proteins to the apical and the basal domains of the plasma membrane are not contained within the primary amino acid sequence. PMID- 7492328 TI - Acyl-CoA binding and acylation of UDP-glucuronosyltransferase isoforms of rat liver: their effect on enzyme activity. AB - When [14C]arachidonoyl-CoA was incubated with crude extracts of rat liver microsomes, [14C]arachidonic acid was incorporated into many proteins, suggesting that modification of these proteins with fatty acid, i.e. acylation, occurred. Using a [14C]arachidonyl-CoA labelling assay, 50 and 53 kDa proteins were purified from rat liver microsomes to near homogeneity by sequential chromatography on Red-Toyopearl, hydroxyapatite, heparin-Toyopearl, Blue Toyopearl and UDP-hexanolamine-agarose. Acylation of the 50 and 53 kDa proteins occurred in the absence of any other protein, suggesting that these molecules catalyse autoacylation. The acylation was dependent on the length of the incubation period and the concentration of [14C]arachidonoyl-CoA. The 50 and 53 kDa proteins also had acyl-CoA-binding activity; initial rates of acyl-CoA binding and acylation were 0.25 and 0.004 min-1 respectively. The proteins also had weak but distinct acyl-CoA-hydrolysing activity (0.006 min-1). These results suggest that the proteins catalysed the sequential reactions of binding to acyl CoA, autoacylation, and hydrolysis of fatty acid. N-terminal amino acid sequencing analysis showed these proteins to be UDP-glucuronosyltransferase (UDPGT) isoforms. UDPGT activity was inhibited by arachidonoyl-CoA. These results suggest that binding of acyl-CoA and acylation of UDPGT isoforms regulate the enzyme activities, implying a possible novel function for fatty acyl-CoA in glucuronidation, which is involved in the metabolism of drugs, steroids and bilirubin. PMID- 7492329 TI - Re-evaluation of the distributions of dystrophin and utrophin in sciatic nerve. AB - Differential expression of proteins belonging to the dystrophin family was analysed in peripheral nerves. In agreement with previous reports, no full-size dystrophin was detectable, only Dp116, one of the short dystrophin products of the Duchenne muscular dystrophy (DMD) gene. We used specific monoclonal antibodies to fully investigate the presence of utrophin, a dystrophin homologue encoded by a gene located on chromosome 6q24. Evidence is presented here of the presence of two potential isoforms of full-length utrophin in different nerve structures, which may differ by alternative splicing of the 3'-terminal part of the utrophin gene according to the specificities of the monoclonal antiobodies used. One full-length utrophin was co-localized with Dp116 in the sheath around each separate Schwann cell-axon unit, but the other utrophin isoform was found to be perineurium-specific. We also highlighted a potential 80 kDa utrophin-related protein. The utrophin distribution in peripheral nerves was re-evaluated and utrophin isoforms were detected at the protein level. This preliminary indication will require more concrete molecular evidence to confirm the presence of these two utrophin isoforms as well as the potential 80 kDa utrophin isoform, but the results strongly suggest that each isoform must have a specialized role and function within each specific nervous structure. PMID- 7492331 TI - Degradation of purines: only ureidoglycollate lyase out of four allantoin degrading enzymes is present in mammals. AB - It is generally accepted that all of the allantoin-degrading enzymes (allantoinase, allantoicase, ureidoglycollate lyase and urease), used in purine degradation, were lost during mammalian evolution. However, surprisingly, ureidoglycollate lyase has been found in a mammalian tissue. Ureidoglycollate lyase was purified to homogeneity and characterized from rat-liver mitochondria. The apparent Km (17 mM) of the rat enzyme for ureidoglycollate was much higher than that (0.33 mM) of fish-liver ureidoglycollate lyase. The rat-liver enzyme differed from the fish-liver enzyme in enzymic, physical and immunological properties. PMID- 7492330 TI - Pseudopregnancy induces the expression of hepatocyte nuclear factor-1 beta and its target gene aminopeptidase N in rabbit endometrium via the epithelial promoter. AB - The rabbit endometrium is an excellent model system allowing experimental manipulation of aminopeptidase N (APN) mRNA expression in vivo. By RNase mapping and sequencing of cloned PCR-amplified primer-extended RNA, it was demonstrated that endometrial APN expression is directed by the epithelial APN promoter and is increased in human-choriogonadotropin-induced pseudopregnancy. Cloning and sequencing of the rabbit APN epithelial promoter revealed conservation of the upstream footprint (UF), hepatocyte nuclear factor-1 (HNF1) and Sp1 elements known to be present in the pig and human promoters as well. The pseudopregnancy induced APN expression was found to be accompanied by a parallel increase in the level of the transcription factor HNF1 beta, whereas a much smaller increase in Sp1 and UF-binding proteins was observed. This indicates that HNF1 beta acts as a switch triggering the pregnancy-induced APN expression. The sequence of the UF element suggests members of the nuclear hormone-receptor superfamily as possible UF-binding proteins, and competition experiments suggest that the chicken ovalbumin upstream promoter transcription factor functions as such in the rabbit endometrium. PMID- 7492332 TI - The accumulation and compartmentalization of isometamidium chloride in Trypanosoma congolense, monitored by its intrinsic fluorescence. AB - Interaction of the trypanocide isometamidium chloride with components of Trypanosoma congolense results in characteristic shifts in the intrinsic fluorescence of the drug. The specificity of this interaction was investigated by analysing the effects of various physicochemical manipulations on its fluorescence properties. The characteristic shifts involved a preferential increase in the intensity of one emission peak over the other, resulting in a systematic increase in the ratio of fluorescence intensities. These effects were apparently due to constraints on fluorophore free rotation in the solution (that is, viscosity). Purified DNA produced similar effects in a saturable manner displaying high affinity for the drug, indicating that the constraint involves binding of the drug to high-affinity binding sites within the DNA. Such binding sites were demonstrated in lysates derived from trypanosomal cells. The binding sites were associated with macromolecular species (M(r) > 12000), and were partly disrupted by thermal denaturation and proteolysis. Treatment with DNase 1 produced high levels of disruption of the binding sites (> 85%), indicating an involvement of DNA in the binding. BSA demonstrated weak non-specific binding of the drug. Entry of drug into live trypanosomal cells (monitored by 14C-labelled drug uptake) was paralleled by fluorescence shifts observed under comparable conditions of drug concentration and buffer conditions. Both systems (fluorescence shifts and accumulation of labelled drug) indicated the presence of a saturable membrane transporter with high affinity for the drug. We conclude that monitoring the fluorescence shifts of isometamidium constitutes a sensitive and highly specific probe for entry of the drug into trypanosomal cells, thereby enabling resolution of the transport events involved. PMID- 7492333 TI - Novel cellulose-binding domains, NodB homologues and conserved modular architecture in xylanases from the aerobic soil bacteria Pseudomonas fluorescens subsp. cellulosa and Cellvibrio mixtus. AB - To test the hypothesis that selective pressure has led to the retention of cellulose-binding domains (CBDs) by hemicellulase enzymes from aerobic bacteria, four new xylanase (xyn) genes from two cellulolytic soil bacteria, Pseudomonas fluorescens subsp. cellulosa and Cellvibrio mixtus, have been isolated and sequenced. Pseudomonas genes xynE and xynF encoded modular xylanases (XYLE and XYLF) with predicted M(r) values of 68,600 and 65000 respectively. XYLE contained a glycosyl hydrolase family 11 catalytic domain at its N-terminus, followed by three other domains; the second of these exhibited sequence identity with NodB from rhizobia. The C-terminal domain (40 residues) exhibited significant sequence identity with a non-catalytic domain of previously unknown function, conserved in all the cellulases and one of the hemicellulases previously characterized from the pseudomonad, and was shown to function as a CBD when fused to the reporter protein glutathione-S-transferase. XYLF contained a C-terminal glycosyl hydrolase family 10 catalytic domain and a novel CBD at its N-terminus. C. mixtus genes xynA and xynB exhibited substantial sequence identity with xynE and xynF respectively, and encoded modular xylanases with the same molecular architecture and, by inference, the same functional properties. In the absence of extensive cross-hybridization between other multiple cel (cellulase) and xyn genes from P. fluorescens subsp. cellulosa and genomic DNA from C. mixtus, similarity between the two pairs of xylanases may indicate a recent transfer of genes between the two bacteria. PMID- 7492335 TI - Effects of hydration state on the synthesis and secretion of triacylglycerol by isolated rat hepatocytes. Implications for the actions of insulin and glucagon on hepatic secretion. AB - The effects of hepatocyte volume on the secretion of triacylglycerol were studied in order to test the suggestion that increases in the portal concentrations of osmolyte amino acids and metal ions during the prandial/early-absorptive phase may be involved in mediating the acute changes in glycerolipid metabolism observed in vivo [Zammit (1995) Biochem Soc. Trans. 23, 506-511]. Incubation of isolated rat hepatocytes with hypo-osmotic medium or in the presence of glutamine (in the presence or absence of leucine), conditions which gave an increase in cell water content of between 8 and 27%, resulted in a decrease in the rate of [14C]triacylglycerol (TAG) secretion when [14C]palmitate was used as substrate. The inhibition was proportional to the increase in cell water content. At low exogenous palmitate concentration (0.05 mM), the inhibition of [14C]TAG secretion was accompanied by a marked shift in the incorporation of label from TAG to phospholipid. In the presence of 0.5 mM palmitate this effect was attenuated, and in the presence of 1 mM palmitate it was abolished. Increased cell volume associated with incubation of hepatocytes with glutamine (in the presence or absence of leucine) also resulted in a decrease in the fraction of newly labelled TAG that was secreted into the medium. Decreased cell volume, achieved by incubation of hepatocytes with hyperosmotic medium (sufficient to decrease cell water content by approx. 9%) decreased overall [14C]TAG secretion, but did not affect the amount of label that was incorporated into phospholipid as a fraction of that incorporated into total glycerolipids. Cell shrinkage, however, diminished the fraction of newly labelled [14C]TAG that was secreted. When intracellular TAG was prelabelled with [3H]glycerol, it was found that cell shrinkage markedly inhibited (preformed) [3H]TAG secretion, whereas cell swelling did not affect this route of TAG secretion. The data are discussed in terms of the possible action of changes in cell hydration at the different loci at which hepatocyte TAG secretion is controlled, with reference to previous observations that both insulin and glucagon are able to inhibit TAG secretion in cultured rat hepatocytes and HepG2 cells. PMID- 7492336 TI - Changes in the components of a nuclear inositide cycle during differentiation in murine erythroleukaemia cells. AB - Differentiation of murine erythroleukaemia cells with the chemical agent DMSO leads to a cessation of proliferation and the production of a number of erythrocyte markers such as haemoglobin. We have previously demonstrated that activation of proliferation leads to an increase in the production of nuclear diacylglycerol (DAG). Here we demonstrate that differentiation leads to a decrease in the levels of nuclear DAG and the activity of the nuclear-associated phosphoinositidase C (PIC). The change in activity appears to be due to a decrease in the mass levels of the beta 1 isoform, as demonstrated by the use of isoform-specific antibodies. Moreover, the changes correlate with the cessation of proliferation and an increase in the number of cells in G1 phase of the cell cycle, rather than with the number of cells which have differentiated. Indeed, although treatment of the cells with phorbol 12-myristate 13-acetate (PMA) inhibits the differentiation programme as assessed by haemoglobin staining, it does not inhibit the number of cells blocking in G1 of the cell cycle or the changes in nuclear DAG or PIC activity. The possible involvement of this nuclear inositide cycle during progression through the cell cycle is discussed. PMID- 7492334 TI - Characterization of des-(741-1668)-factor VIII, a single-chain factor VIII variant with a fusion site susceptible to proteolysis by thrombin and factor Xa. AB - A factor VIII variant has been characterized in which the heavy chain is directly fused to the light chain. Des-(741-1668)-factor VIII lacks the processing site at Arg1648, as Arg740 of the heavy chain is fused to Ser1669 of the light chain. The sequence of the fusion site is similar to that of other cleavage sites in factor VIII. The fusion site of des-(741-1668)-factor VIII was readily cleaved by both thrombin and factor Xa, and the same result was obtained for heavy chain cleavage. In contrast, des-(741-1668)-factor VIII cleavage by thrombin at position Arg1689 proceeded at a lower rate than the analogous cleavage by factor Xa, which presumably takes place at position Arg1721. The rate of cleavage at position Arg1689 by thrombin was also lower than that at the other processing sites. When des-(741-1668)-factor VIII was activated by thrombin, initial rates of factor Xa formation were similar to the rates obtained when plasma-derived factor VIII was activated by thrombin or factor Xa. Remarkably, activation of des (741-1668)-factor VIII proceeded at a higher rate by factor Xa than by thrombin. These results indicate that factor VIII activation is strongly associated with cleavage at position Arg1689 or Arg1721. For the interaction between des-(741 1668)-factor VIII and von Willebrand factor, a Kd value of (0.8 +/- 0.3) x 10( 10) M was determined, which is similar to that of heterodimeric factor VIII. The affinity of single-chain des-(741-1668)-factor VIII for factor IXa was found to be 27 +/- 6 nM. The in vivo recovery and half-life of des-(741-1668)-factor VIII were assessed in guinea pigs. Upon infusion of des-(741-1668)-factor VIII at a dosage of 50 units/kg body weight, a rise of 1.0 +/- 0.3 unit/ml in factor VIII activity was obtained. The same recovery was determined for wild-type factor VIII. The half-life of des-(741-1668)-factor VIII was found to be 3 +/- 1 h, compared with 4 +/- 2 h for heterodimeric recombinant factor VIII. In conclusion, des-(741-1668)-factor VIII displays normal activity, is readily cleaved by thrombin and factor Xa at its fusion site, binds with high affinity to von Willebrand factor and factor IXa, and behaves like heterodimeric recombinant factor VIII in guinea pigs. By virtue of these properties, des-(741-1668)-factor VIII may prove useful for the treatment of bleeding episodes in patients with haemophilia A. PMID- 7492337 TI - Molecular evidence for the direct involvement of a protein kinase C in developmental and behavioural susceptibility to tumour-promoting phorbol esters in Caenorhabditis elegans. AB - The nematode Caenorhabditis elegans displays developmental and behavioural sensitivity to tumour-promoting phorbol esters. This sensitivity involves the gene tpa-1, which encodes two protein kinase C isoforms, TPA-1A and TPA-1B. Here we report the molecular nature of the sensitivity in this animal. Characterization of transposon Tc1-induced phorbol ester-resistant mutants has revealed that Tc1 was inserted in a region encoding the kinase domain, resulting in the loss of tpa-1 products. Introduction of a genomic DNA containing the entire wild-type tpa-1 locus into a Tc1-inserted mutant restored the sensitivity to tumour promoters, and tpa-1 products were also produced. These results suggest that the function of wild-type TPA-1 is necessary and sufficient for tumour promoters to cause developmental and behavioural sensitivity in C. elegans. PMID- 7492338 TI - Metabolism of [3-13C]pyruvate and [3-13C]propionate in normal and ischaemic rat heart in vivo: 1H- and 13C-NMR studies. AB - The oxidation of [3-13C]pyruvate and [3-13C]propionate was studied in vivo in infused rats. The infused [3-13C]pyruvate was quickly converted to [3-13C]lactate in the blood, and the [3-13C]lactate formed was well metabolized in both normoxic and ischaemic hearts. Large differences (200-600%) in the 13C enrichment of alanine (C-3) and acetyl-CoA (C-2) compared with lactate (C-3) were found in both normoxic and ischaemic hearts, suggesting that the extracellular [3-13C]lactate preferentially entered a region of the cytoplasm which specifically transfers the labelled pyruvate (formed from [3-13C]lactate) to the mitochondria. The highly enriched mitochondrial pyruvate gave high enrichment in alanine and acetyl-CoA, which was detected by 1H- and 13C-NMR spectroscopy. Ischaemia increased 13C incorporation into the main cytoplasmic lactate pool and decreased 13C incorporation into citric acid cycle intermediates, mainly decreasing the pyruvate anaplerosis. Isoprenaline-induced ischaemia of the heart caused only a slight decrease in pyruvate oxidation. In contrast to the decreased anaplerosis of pyruvate, the anaplerosis of propionate (and propionyl-carnitine) increased significantly in ischaemic hearts, which may contribute to the protective effect of propionyl-carnitine seen in ischaemia. In addition, we found that [3 13C]propionate preferentially labelled aspartate C-3 in rat heart, suggesting incomplete randomization of label in the succinyl-CoA-malate span of the citric acid cycle. These data show that proton observed 13C edited spectroscopic methods, i.e. heteronuclear spin-echo and the one-dimensional heteronuclear multiple quantum coherence sequence, can be successfully used to study heart metabolism in vivo. PMID- 7492340 TI - Amino acid sequencing, molecular cloning and modelling of the chick liver class theta glutathione S-transferase CL1. AB - Glutathione S-transferase CL1-2 heterodimers purified from 1-day-old chick livers were digested with Achromobacter proteinase I. The resulting fragments were separated for amino acid sequence analysis. Oligonucleotide probes were constructed based on sequence similarity to class-Theta glutathione S transferases for PCR using a chicken liver cDNA library as template. A full length clone (1725 bp) encoding a polypeptide comprising 261 amino acids was isolated. Including conservative substitutions, this protein has 70-73% sequence similarity with other mammalian class-Theta glutathione S-transferases. Based on known X-ray crystal structures of class-Alpha, -Mu and -Pi glutathione S transferases, a model is constructed for the N-terminal 232 residues of CL1. PMID- 7492339 TI - A new model for disruption of the ornithine decarboxylase gene, SPE1, in Saccharomyces cerevisiae exhibits growth arrest and genetic instability at the MAT locus. AB - Ornithine decarboxylase (ODC) is a rate-determining enzyme of the polyamine biosynthetic pathway. We sought to produce cells with impaired ODC function in order to study the biological functions of polyamines. Saccharomyces cerevisiae strains were obtained by one-step gene replacement of a 900 bp fragment of the yeast ODC gene (SPE1) with the yeast URA3 gene. Spores derived from SPE1/spe1 cells germinated at reduced efficiency relative to SPE1/SPE1. Sustained growth of spe1 haploid mutants in polyamine-free medium led to intracellular polyamine depletion, reduction in budding index, G1 arrest and cessation of growth, and cells that were large and misshapen. All of these effects were completely reversed by adding polyamines to the medium, even after 5 days of polyamine starvation. A diploid yeast strain bearing two copies of disrupted spe1 lost heterozygosity at the mating-type locus more often when grown in the absence of polyamines than when grown in their presence, indicating that polyamine deficiency leads to either chromosome loss or to mitotic recombination. PMID- 7492342 TI - European Respiratory Society annual congress. Barcelona, Spain, September 16-20, 1995. Abstracts. PMID- 7492343 TI - Insulin-dependent diabetes mellitus as a beta-cell targeted disease of immunoregulation. PMID- 7492341 TI - The nature of topogenic sequences determines the transport competence of topological mutants of neutral endopeptidase-24.11. AB - Type II integral membrane proteins are anchored by a signal-peptide/membrane anchor domain (SA domain) located near their N-terminus, whereas type I membrane proteins are anchored by stop-transfer sequences usually located near the C terminus. In this study we have attempted to transform neutral endopeptidase 24.11 (EC 3.4.24.11; NEP), a type II membrane protein, into a type I membrane protein. Three type I mutant proteins were constructed by fusion of topogenic sequences to the C-terminus of SecNEP, a soluble form of NEP. The first two type I mutants, SecNEP-TMC and SecNEP-TMIC, were constructed by fusing in frame the cytosolic and SA domains of NEP to the C-terminus of SecNEP. These two fusion proteins differ only in the orientation of the cytosolic tail. The third type I mutant, SecNEP-ACE, was constructed by fusing in frame the stop-transfer and cytosolic domains of angiotensin I-converting enzyme (EC 3.4.15.1; ACE) to the C terminus of SecNEP. Our results suggest that: (1) the NEP ectodomain can be anchored with a type I topology in the endoplasmic reticulum (ER) membrane by both NEP and ACE topogenic sequences; (2) SecNEP-TMC and SecNEP-TMIC were transport-incompetent and needed proteolytic cleavage in the C-terminal region to leave the ER, whereas SecNEP-ACE was transported out of the ER as a type I membrane protein. Therefore we concluded that the nature of topogenic sequences determines the transport-competence of topological mutants of neutral endopeptidase-24.11. PMID- 7492344 TI - Pancreatic expression of antigens for islet cell antibodies in non-obese diabetic mice. AB - Diabetes-prone NOD mice of both sexes and at different ages were compared to control mice with regard to the level of pancreatic expression of certain autoantigens: antigens for islet cell antibodies (ICA antigens) and glutamic acid decarboxylase (GAD) 67 kDa. ICA antigens were compared by immunofluorescence using serial dilutions of ICA positive human sera so that differences of fluorescence intensity were due only to differences in amounts of antigen. Pancreatic GAD67 mRNAs were compared by polymerase chain reaction followed by Southern hybridization with 32P-probes and densitometry of autoradiographic bands. GAD67 product and gamma-aminobutyric acid (GABA) were compared by immunoperoxidase staining. As compared to BALB/c, C57BL6, Swiss, or F1 mice, NOD mice displayed higher ICA antigen levels (P < 0.01) both before and after insulitis onset (at 7 days, 15 days, 1 month, 2 months). ICA antigens were scarcely detectable by the first day of life, and increased with age from 7 days to 2 months (P < 0.01; n = 10 for each strain and at each age). Both before and after insulitis onset (4 days, 7 days, 15 days, 1 month, 2 months), amounts of GAD67 mRNAs were higher (P < 0.01) in NOD mice than in BALB/c mice (n = 8 for each age in each strain). This was already noted in foetuses on Day 18 of gestation (n = 8). After birth, amounts of GAD67 mRNAs increased up to 1 month (P < 0.04) and then decreased in older mice. The staining intensity of pancreatic sections with antisera against either GAD67 or GABA was higher (P < 0.04) in islets from NOD mice than in those from control mice. Whatever the age, no significant difference was noted between female and male NOD mice with regard to ICA antigens or GAD67. The expression of ICA antigens and GAD67 was intermediate in NOD x BALB/c F1 mice when compared to parental strains. We conclude that whatever the age, NOD mice strongly express ICA antigens and GAD67. This peculiarity was detectable very early, in embryos for GAD67 but after birth for ICA antigens. The timing of antigen expression may underlie the development of diabetes. The antigen overexpression might affect early completion of self tolerance and, during later life, might also contribute to amplification of the anti-beta cell autoimmune response due to the existence of more targets for effector mechanisms. PMID- 7492347 TI - Preferential TCR V beta 1 gene usage by autoreactive T cells in spontaneous autoimmune thyroiditis of the obese strain of chickens. AB - We studied T cell receptor variable beta (TCR V beta) gene usage by autoreactive T cells in spontaneous autoimmune thyroiditis (SAT) of obese strain (OS) chickens. Chicken alpha beta T cells may express either V beta 1 or V beta 2 genes, the products of which can be recognized by TCR2 and TCR3 monoclonal antibodies, respectively. Selective depletion of V beta 1+ or V beta 2+ T cells in OS chickens was accomplished by repeated injections of TCR2 or TCR3 antibodies into embryonic and 1-3-week-old chickens. The birds were killed at 20 days of age and their spleens and thyroid glands evaluated by immunohistochemistry. We found that V beta 1+ T cells preferentially infiltrated OS chicken thyroid glands. Antibody treatments resulted in a 41% reduction in frequency of V beta 1+, and a 87% reduction of the frequency of V beta 2+ cells in the circulation, and in a profound decrease of the respective T cells in spleens and thyroid glands. Selective suppression of V beta 1+ T cells partially inhibited SAT development in that thyroid-infiltrating cells and destruction of thyroid follicles were reduced by more than 50%. Thyroglobulin autoantibody serum levels were also reduced in V beta 1+ T cell-depleted OS chickens, whereas selective depletion of V beta 2+ T cells did not inhibit SAT development. These findings indicate preferential TCR V beta 1 gene usage by autoreactive T cells in SAT of OS chickens. PMID- 7492345 TI - The role of infiltrating macrophages in islet destruction and regrowth in a transgenic model. AB - The expression of interferon-gamma (IFN-gamma) in pancreatic beta cells leads to a complex pathology that represents the processes of both islet destruction and islet regeneration. Inflammatory cells and the factors elicited from them participate in the development of pathology in this transgenic model. To dissect the role of infiltrating macrophages in these events, the monoclonal directed against the type 3 complement receptor (5C6) was utilized to inhibit the extravasation of macrophages. This was approached by treating transgenic mice with 5C6 for 3 or 4 months, starting from 5-7 days of age. The data presented in this report demonstrate that infiltrating macrophages are important in the observed induction of diabetes in our transgenic model. We also found that infiltrating macrophages did not play a major role in the observed proliferation and islet regeneration, but some interesting subtleties regarding the regulation of this proliferative process emerged. PMID- 7492346 TI - Thyroiditis in the BB rat is associated with lymphopenia but occurs independently of diabetes. AB - The spontaneously diabetic BB rat is an excellent and well studied model for human insulin-dependent diabetes (IDDM), sharing many important features with the human disease. Similarities include an equal frequency of IDDM in males and females, production of antibodies against pancreatic cell antigens, and an MHC disease association. In addition, the BB rat shares with human IDDM patients an increased frequency of autoantibodies against the parietal cells of the stomach and colloid cells of the thyroid gland. Here we investigate the genetic basis of thyroiditis in the BB rat. Based on crosses between BB, Lewis and Fischer rats, we show that two susceptibility factors for diabetes--the lymphopenia trait present in diabetes prone BB rats and the MHC--also appear to be risk factors for thyroiditis. However, the nature of the susceptibility was different for the two autoimmune diseases, with lymphopenia being absolutely required for diabetes although it only conferred increased risk for thyroiditis. Also, in contrast to IDDM, the MHC conferred dominant susceptibility to thyroiditis. Despite these shared risk factors, diabetes per se did not show significant correlation with thyroiditis. PMID- 7492348 TI - Expression and regulation of anti-thyroid autoimmunity directed against cultivated rat thyrocytes. AB - Lymph node cells from DA rats that had been exposed in utero to 131I in doses sufficient to interrupt thyroid development, attacked monolayers of normal syngeneic thyrocytes in vitro. Lymph node cells from normal DA rats did not damage syngeneic thyrocyte monolayers. Thyrocytes could be protected from damage provided they had been incubated with lymph node cells from normal syngeneic rats before the introduction of lymph node cells from 131I exposed rats. Spleen cells from both 131I exposed and normal rats attacked syngeneic thyrocytes. It is concluded that normal rats possess cells capable of downregulating anti-thyroid autoimmunity. PMID- 7492349 TI - Suppression of anti-thyrocyte autoreactivity by the lymphocytes of normal fetal lambs. AB - We have devised an experimental strategy to determine whether the developing immune system of normal fetal animals can spontaneously acquire the capacity to inhibit autoimmune responses by its cells as it matures. Whilst the existence of cells with the capacity to exert negative regulation and to curtail autoimmune responses has been demonstrated previously in response to the experimental induction of these responses, the relevance of such regulatory processes to the prevention of overt autoimmunity in normal animals has not been established. We have produced pairs of identical twin fetal lambs by splitting blastocysts and have subsequently deprived one of each pair of exposure to thyroid-specific antigens by surgical thyroidectomy before development of immunological self recognition. Thyroidectomized fetuses developed T lymphocytes autoreactive against self thyrocytes. However, their normal, identical co-twins were found to acquire a class of T lymphocytes with the capacity to block anti-thyrocyte autoreactive cells from the thyroidectomized fetal co-twin. Blocking of anti thyroid autoreactivity required preliminary contact between these normal T lymphocytes and the target thyrocytes. Substitution of an allograft of fetal thyroid tissue for a fetal lamb's own thyroid gland failed to prevent the development of autoreactivity against autologous thyrocytes by the recipient's lymphocytes. However, the reactivity of those lymphocytes against thyrocytes from the specific allogeneic thyroid donor was markedly curtailed. PMID- 7492350 TI - Lack of proliferative response by gluten-specific T cells in the blood and gut of patients with dermatitis herpetiformis. AB - The majority of patients with Dermatitis Herpetiformis (DH) have a gluten sensitive enteropathy which may be triggered by a T cell-mediated immune response to gluten. Using a proliferative assay, the responses to gluten fraction III, recall antigens and mitogens of peripheral blood mononuclear cells (PBMC) and gut T cell lines (TCL) isolated from patients with Dermatitis Herpetiformis (DH) and normal controls were studied. In most cases, neither PBMC nor gut T cell lines (which were predominantly CD3+, CD4+, TCR alpha beta +) from either controls or patients proliferated in response to gluten fraction III alone. However, the addition of 10 U/ml IL-2 to PBMC cultures containing gluten fraction III resulted in a marked increase in proliferation in 9/19 DH patients and 7/11 controls compared to IL-2 alone. Furthermore, gluten-induced upregulation of IL-2 receptor (CD25) expression was demonstrated on PBMC from 4/4 patients with DH and 2/3 controls after 7 days' culture with antigen. A similar effect by exogenous IL-2, or the same concentration of IL-4, was observed in 8/11 (P = 0.02) and 5/6 respectively DH, and 3/4 normal gut T cell lines. No difference was observed in the response of DH and control PBMC to Tetanus toxin, Candida albicans and PPD; both normal and DH gut T cell lines were unresponsive to these antigens. However, the addition of IL-2 increased the response to Candida albicans by DH gut T cell lines. Moreover, the response of DH gut T cell lines to PHA (P < 0.001), Concanavalin A and anti-CD3 were markedly reduced compared to PBMC from the same patients. These findings suggest that gluten-specific T cells present in the blood and gut of normal and DH individuals are activated by but do not proliferate in response to specific antigen. PMID- 7492351 TI - Autoimmunity to the cell cycle-dependent centromere protein p330d/CENP-F in disorders associated with cell proliferation. AB - p330d/CENP-F is a novel proliferation-associated and cell cycle-dependent centromere autoantigen which appears to play a very important role in mitotic progression. As an initial step in exploring the clinical and biological significance of autoantibodies to this protein, we evaluated the clinical histories of 26 patients producing these antibodies. The antibodies were detected by both indirect immunofluorescence microscopy (IIF) and Western blotting. All the sera contained anti-p330d/CENP-F IgG antibodies, with an average titer by IIF of 1:6,917 (range 1:160 to 1:20,480). Most of the patients had disorders associated with abnormal or increased cell proliferation at the time the anti p330d/CENP-F antibodies were detected. These included cancers of various types (14), chronic liver disease (3), chronic rejection of renal allografts (2), and Crohn's disease (1). The average IIF titer of the anti-p330d/CENP-F antibodies in the patients with cancer, 1:10,103, was significantly higher than the average titer in non-cancer patients, 1:3,200 (P = 0.008). Autoimmunity to p330d/CENP-F appeared not to be associated with rheumatic diseases, in particular scleroderma, since only three of the 26 patients had rheumatic disease and the antibodies were not detected by IIF in a group of 351 patients with scleroderma and related disorders. Our findings, although retrospective and limited to a relatively small number of patients, point to the hypothesis that autoimmunity to p330d/CENP-F could be related to events involving increased or abnormal cell proliferation. PMID- 7492352 TI - Upregulated expression of IL-4 receptors and increased levels of IL-4 in rheumatoid arthritis patients. AB - The level of IL-4R expression on peripheral lymphocyte subsets from rheumatoid arthritis (RA) patients and controls was studied by flow cytometric analysis of the binding of phycoerythrin-labelled IL-4 (PE-IL-4). In normal lymphocytes, IL 4R is mainly expressed on CD19+ cells, although it was also seen, at lower levels, on CD3+, CD4+ and CD8+ cells. In RA patients, a significantly increased spontaneous expression of IL-4R was observed, compared with controls, in the CD3+, CD4+ and CD19+ cell subsets. No significant differences in IL-4R expression were found between patients receiving steroids and those who were not, suggesting that steroids are not involved in upregulating IL-4R levels in vivo. Because IL-4 is a potent upregulator of IL-4R, we considered the possibility that incremented levels of circulating IL-4 in RA accounted for the high surface expression of IL 4R. By ELISA, we found abnormally high levels of immunoreactive IL-4 in 35.13% of patient serum samples, while it was undetectable in control sera. In addition, we examined IL-4 mRNA expression by polymerase chain reaction (PCR) in the PBMC of patients and controls. IL-4 PCR products were observed in four out of 10 patients studied but in none of the controls. No correlation was observed between the seric concentrations of IL-4 and IL-4R, indicating that activator factors other than IL-4 contribute to the upregulation of IL-4R expression in RA. Since the patients' sedimentation rate and CRP values did not correlate with the concentration of circulating IL-4, we conclude that this lymphokine does not contribute to the deleterious effect of the disease. Rather, due to its antiinflammatory properties, the overproduction of IL-4 in RA may be a compensatory mechanism neutralizing the harmful effect of activated macrophages. PMID- 7492355 TI - [New theoretical study on the biological origins of Chinese medicinal herbs]. AB - Following the theses on "A theoretical study of the biological origins of Chinese medicinal herbs" in 1991, the authors continued the study and established eleven new views, namely; "The theory of differentiation tendency of Chinese medicinal herbs for regional habitually used drugs"; "The theory of regularity of occurrence of confusing species and pseudo materia"; "Discussion on rich resources of new drugs from old sorts of traditional Chinese medicine"; "The theory of knowledge of substitutes for Chinese medicinal herbs", etc. PMID- 7492353 TI - Antigenic specificity of lymphocytes isolated from valvular specimens of rheumatic fever patients. AB - T cell lines were established from both valvular specimens and peripheral blood lymphocytes from seven patients with well documented rheumatic heart disease. These cell lines were stimulated with either PHA or streptococcal antigens. Proliferation assays revealed that both valvular and peripheral blood T cell lines reacted to cell wall (CW) and cell membrane (CM) antigens obtained from rheumatic fever associated group A streptococci and not to nephritogenic strains. None of the cell lines reacted to M protein, myosin or other mammalian cytoskeletal proteins. The unique reactivity of rheumatic fever T cell lines only to cellular structures obtained from rheumatogenic strains suggests that these lines react to epitopes specific for antigens obtained from these strains. PMID- 7492354 TI - Activation and clonal expansion of human myelin basic protein-reactive T cells by bacterial superantigens. AB - Autoreactive T cells specific for myelin basic protein (MBP) are part of the normal T cell repertoire and are present both in patients with multiple sclerosis (MS) and healthy individuals. There is evidence suggesting in vivo activation and persistent clonal expansion of MBP-reactive T cells in MS. This study was undertaken to investigate the potential role of bacterial superantigens (SA) in the activation of MBP-reactive T cells. Twenty-seven MBP-reactive T cell clones generated from 10 MS patients and one normal individual were examined for reactivity to SA, in association with their T cell receptor V beta gene usage. The majority of the clones responded to at least one of the SA tested, staphylococcal enterotoxins (SEA and SEB) and toxic shock syndrome toxin-1 (TSST 1). The clones reactive to SEA and SEB expressed various V beta genes while T cell reactivity to TSST-1 correlated with the V beta 2 expression. Furthermore, circulating MBP-reactive T cells could be expanded from lymphocyte cultures primarily exposed to respective SA in more than 50% of MS patients and normal individuals tested. However, activation and expansion of circulating MBP-reactive T cells by SA was not directly associated with the disease. This study lends support to the potential role of SA in the activation of MBP-reactive T cells and suggests that an altered regulatory mechanism may account for further expansion and persistence of MBP-reactive T cells in MS. PMID- 7492356 TI - [Effect of processing on specific toxicity of radix Achyranthis bidentatae]. AB - To activate Epstein-barr virus in Raji cells in vitro the lowest activation concentrations of ether extracts from Radix Achyranthis Bidentatae unprepared (I), stir-baked with wine (II) and stir-baked with salt (III) required were 0.8.20 and 0.16 micrograms/ml respectively. 5 microliters (equal to 250 mg herb weight)/ear of these extracts had inflammatory effect on mouse ear skin in 4 hours. Ethanol extract of I at concentration 0.04-125 mg/ml did not increase the chromosomal aberration rates in CHL cells. The decoctions of I, II, and III given orally at 15 g/kg x 3d showed no effect on micronucleus rates in mouse bone marrow. Ethanol extracts of I, II, and III at 10 g/kg x 10d did not show any evident antifertility action. PMID- 7492357 TI - [Effect of unprepared and calcined magnetitum of arsenic solubility in boiling water extracts]. PMID- 7492358 TI - [Comparative study on baicalin contents in 4 traditional Chinese medicinal preparations of ying huang]. AB - The contents of baicalin in four different traditional Chinese medicinal preparations of Ying Huang were determined by HPLC. The contents of baicalin in Ying Huang tablets, Ying Huang oral liquid, Ying Huang injection and Ying Huang granules are respectively: 124.18 mg/g, 21.12 mg/ml, 12.39 mg/ml and 6.79 mg/g. The HPLC method suggested in this paper can be used for quality control in the production of Ying Huang preparations. PMID- 7492359 TI - [Analysis of poisonous trace elements in huangwu gongmitai]. AB - A comparative study was made on the dissolved amount of poisonous trace elements mercury and arsenic contained in hydrargyri subchloridum and realgar of gynecological suppository in artificially simulated vaginal acid-base liquid with that contained in hydrargyri subchloridum and realgar in artificial gastric juice stipulated in Pharmacopeia of China. The result shows that the dissolved amount of mercury and arsenic contained in the suppository is much smaller than that stipulated in Pharmacopeia for oral administration. PMID- 7492360 TI - [Chemical components of Seseli yunnanense Franch]. PMID- 7492361 TI - [Chemical constituents in the leaves of Mangifera persiciformis C.Y. Wu et Y.L. Ming]. AB - Eleven crystalline constituents have been isolated from the leaves of Mangifera persiciformis, of which five were identified as taraxerol, friedelin, beta sitosterol, mangiferin and quercetin by comparing their physicochemical and spectroscopic data. They were isolated from this plant for the first time. PMID- 7492363 TI - [Chemical constituents of stems and leaves of Fritillaria ussuriensis Maxim]. AB - Three compounds were isolated from stems and leaves of Fritillaria ussuriensis. They were identified as adenosine, galacitiol and succinic acid. PMID- 7492362 TI - [Chemical constituents of Astragalus chinensis L]. AB - Three compounds have been isolated from the seeds of Astragalus chinensis. Their structures were identified on the basis of spectral data (UV, 13CNMR, 1HNMR, IR and MS) as octacosane, daucosterol and kaempferol. PMID- 7492364 TI - [Liver-protecting and alanine aminotransferase lowering actions of hongning gantai]. AB - Hongning Gantai has a remarkable counteraction on the elevation of ALT activity of mice and rats induced by chemical damages with CCl4 and D-Gal. It can decrease the BSP-retention. The result of pathologic examination indicates that Hongning Gantai can alleviate liver damages of mice and rats and thus has liver-protecting and ALT-lowering actions. PMID- 7492365 TI - [Preliminary studies on the desmutagenic mechanism of white silk fowl's melanin]. PMID- 7492367 TI - [Pharmacological research on antiasthmatic liquid for oral administration]. AB - The Oral Antiasthmatic Liquid, when given into the mice stomach, can obviously reduce the times of cough induced by sulfur dioxide aerosol, and increase the amount of phenolsulfonphthalein excretion of tracheas. It also shows antagonistic action to bistamin phosphate and acetylcholine that can contract the isolated tracheas of guinea-pigs, and reduce the amounts of isolated lung and bronchial perfusion of guinea-pigs. PMID- 7492366 TI - [Protective action of Lycium barbarum L. (LbL) and betaine on lipid peroxidation of erythrocyte membrane induced by H2O2]. AB - H2O2 was used to induce lipid peroxidation of RBC membrane in rats to observe the protective action of different ingredients of LbL and betaine on the membrane. The result shows these ingredients cna inhibit the lipid peroxidation of RBC membrane in the following order of antioxidizing power: Fructus Lycii.LbL.dry(FL/LbL.dry) > Polysaccharide FL/LbL > Residue FL/LbL > betaine. PMID- 7492368 TI - [Advances in the research of liuwei dihuang decoction]. PMID- 7492369 TI - Steroidal saponins from Allium chinense and their inhibitory activities on cyclic AMP phosphodiesterase and Na+/K+ ATPase. AB - The saponin fraction prepared from the methanolic extract of Allium chinense bulbs exhibited inhibitory activities on cyclic AMP phosphodiesterase (cAMP PDE) (43.5%) and Na+/K+ ATPase (59.3%) at a sample concentration of 100 micrograms ml 1, respectively. Attempted purification of the active fraction through column chromatography on silica gel and ODS silica gel resulted in the isolation of six steroidal saponins, one of which appeared to be a new compound and one to be the first isolation from a natural source. (25R,S)-5 alpha-Spirostan-3 beta-ol tetrasaccharide showed inhibitory activities on both cAMP PDE and Na+/K+ ATPase, while (25R)-3 beta-hydroxy-5 alpha-spirostan-6-one di- and tri-saccharides inhibited only cAMP PDE. PMID- 7492370 TI - Cyanogenic and non-cyanogenic glycosides from Manihot esculenta. AB - In addition to lotaustralin and linamarin, a novel cyanogenic glycoside, 2-((6-O (beta-D-apiofuranosyl)-beta-D-glucopyranosyl)oxy)-2-met hylbutanenitrile , two novel non-cyanogenic glycosides, (2S)-((6-O-(beta-D-apiofuranosyl)-beta-D- glucopyranosyl)oxy)butane and 2-((6-O-(beta-D-apiofuranosyl)-beta-D glucopyranosyl)oxy)propane, and a simple non-cyanogenic glycoside, ethyl beta-D glucopyranoside, were isolated from an ethanolic extract of the fresh root cortex of Manihot esculenta. From a methanolic extract of the fresh leaves of this species lotaustralin and linamarin, and two flavonoid glycosides, kaempferol-3-O rutinoside and quercetin-3-O-rutinoside were isolated. PMID- 7492371 TI - Oleanene glycosides from seeds of Trifolium alexandrinum. AB - From the seeds of Trifolium alexandrinum, two new oleanene-type triterpene glycosides were isolated as their methyl esters, together with five known saponins. The structures of the isolated compounds were determined by NMR and mass spectral analyses. PMID- 7492372 TI - Antifungal steroidal lactones from Withania coagulance. AB - Two new withanolides, 14,15 beta-epoxywithanolide I [(20S,22R) 17 beta, 20 beta dihydroxy-14 beta,15 beta-epoxy-1-oxo-witha-3,5,24-trienolide] and 17 beta hydroxywithanolide K (20S,22R) 14 alpha,17 beta,20 beta-trihydroxy-1-oxo-witha 2,5,24-trienolide] have been isolated from the whole plant of Withania coagulance and their structures were elucidated by spectroscopic techniques. The latter compound was found to be active against a number of potentially pathogenic fungi. PMID- 7492373 TI - Acylated cyanidin glycosides in the violet-blue flowers of Ipomoea purpurea. AB - Six acylated cyanidin glycosides were isolated from violet-blue flowers of Ipomoea purpurea. These anthocyanins were all based on cyanidin 3-sophoroside-5 glucoside, acylated with caffeic acid and/or p-coumaric acid. Three anthocyanin structures were elucidated to be cyanidin 3-O(-)[2-O-(6-O-(trans-3-O-(beta-D glucopyranosyl)caffeyl)-beta -D-glucopyranosyl)-6-O-(trans-4-O-(6-O-(trans caffeyl)-beta-D-glucopyran osyl) caffeyl)-beta-D-glucopyranoside]-5-O(-)[beta-D glucopyranoside], cyanidin 3-O(-)[2-O-(trans-3-O-(beta-D-glucopyranosyl)caffeyl) beta-D-glucopyrano syl)- 6-O-(trans-caffeyl)-beta-D-glucopyranoside]-5-O(-)[beta D-glucopyranosid e d and cyanidin 3-O(-)[2-O-(6-O-(trans-caffeyl)-beta-D glucopyranosyl)-6-O- (trans-caffeyl)-beta-D-glucopyranoside]-5-O(-)[beta-D glucopyranoside]. These three anthocyanins were present in all 12 violet-blue flower strains as major pigments. The colours of these acylated antocyanins were stabler in neutral solution than their deacyl analogues. PMID- 7492374 TI - Lycorine alkaloids from Hymenocallis littoralis. AB - From Hymenocallis littoralis, one new alkaloid, named littoraline, together with 13 known lycorine alkaloids and one lignan, were isolated. The structure and NMR assignments of this new alkaloid were determined by 1D and 2D NMR techniques. Littoraline showed inhibitory activity of HIV reverse transcriptase, and lycorine and haemanthamine showed potent in vitro cytotoxicity. PMID- 7492376 TI - Cancer survival in Sweden during three decades, 1961-1991. AB - Cancer survival in Sweden in 1961-1991 is presented as a comprehensive report from the Swedish Cancer Registry. The report shows both successes and failures, confirms some earlier published results and presents some new findings worth further analysis. Survival has increased for female breast cancer, malignant melanoma, cancers of the testis and thyroid gland, acute leukemia, and Hodgkin's disease. No improvements are found for multiple myeloma or cancers of the liver, gall bladder, and pancreas. Small increases are shown for colorectal cancer and cancers of the stomach, oesophagus, and kidney. Increases in postoperative survival are shown for sites dominated by histologically benign tumors, i.e., intracranial neurinoma, meningioma, and cancers of the endocrine glands such as parathyroid tumors. From 1970-1972 to 1980-1982 the 10-year relative survival rate (RSR) increased from 30% to 38% for males and from 44% to 51% for females. Hence, cancer survival for all cases combined has approached the survival of the general population somewhat. Most of the increases took place in the 1970's. Changes in the distribution of incidence towards cancer sites with better prognoses account for some 10-20% of the observed increases in RSR, whereas the aging of the cancer population reduces the upward trend in RSR for all cases combined by some 1-2%. Cancer patients have poorer survival than the population long after 5 years of follow-up. They reach the survival of the population after about 8-12 years for colorectal cancer, 10 years for cervical cancer, 7-10 years for malignant melanoma, 13-18 years for kidney cancer, and more than 19 years for female breast and prostate cancer. For patients diagnosed in 1970-1972 this occurred 16 years after diagnosis at 29% for males and 43% for females when all cancer cases were combined. The extended time until 'statistical cure' for most cancer forms clearly indicates the need to augment the commonly used 5-year RSR with other outcome measures. If cancers on average are discovered earlier today, the 5-year RSR gives an exaggerated impression of the improvement over time. In this case the change in the 10-year RSR is a less biased criterion. PMID- 7492375 TI - A 4-methyl-7-hydroxyphthalide glycoside and other constituents from Quillaja saponaria molina. AB - A so far unknown 4-methyl-7-hydroxyphthalideglycoside has been isolated from the methanol extract of the bark of Quillaja saponaria molina. Its structure has been established from NMR experiments as 7-O(-)[beta-glucopyranosyl-(1-->6)-beta arabinopyranosyl]-7-hydrox y-4-methy l -1[3H]-isobenzofuranone. Two known compounds, 3,4,5-trimethoxyphenyl-beta-D-glucopyranoside and lyoniresinol-3 alpha O-beta-D-glycopyranoside were also identified. PMID- 7492377 TI - Use of non-proven therapies. Differences in attitudes between Norwegian patients with non-malignant disease and patients suffering from cancer. AB - A comparative study was conducted between a group of patients with non-malignant diseases in general practice and a group of cancer patients seen in the Department of Oncology at the University Hospital of Tromso. The aim of the study was to investigate the prevalent use of 'alternative medicine', here called non proven therapies (NPTs), among cancer patients and general practice patients, and to investigate whether there are any differences in opinion between the two groups regarding the beneficial effects of NPTs as treatment modalities for cancer. A total of 305 general practice patients and 252 cancer patients were included in the final analysis. In both groups close on 20% had been or were present users of NPTs. Among cancer patients the most preferred NPTs methods were healing by laying on of hands and faith healing. The patients with non-malignant disease expressed a more positive view on the possible benefits of NPTs in the fight against cancer than that expressed by the cancer patients. A total of 63.4% of patients from general practice stated that NPTs ought to be an option for cancer patients within Norwegian hospitals. PMID- 7492378 TI - Surgery for cure followed by chemotherapy in small cell carcinoma of the lung. For the ISC-Lung Cancer Study Group. AB - The Lung Cancer Study Group of the International Society of Chemotherapy (ISC LCSG) organized multinational, cooperative, prospective and randomized trials for the cure of patients with small cell lung cancer at early stages (T1,2N0M0). Surgery for cure was used first, followed by postoperative chemotherapy, and thereafter by prophylactic cranial irradiation. Eight cycles of standard chemotherapy (CAV-cyclophosphamide-doxorubicin-vincristine) or 6 intermittent cycles of alternating chemotherapy, using 3 different drug combinations, were administered 1-2 weeks postoperatively for 6 months after randomization. A total of 183 evaluable patients received surgery for cure at 23 cooperating hospitals. The preliminary evaluation of ISC-studies I and II per May 1993 resulted in the 30-month total survival of 63% from 68 patients after complete resection at TN0M0R0-stages and 37% from 27 patients after such resections at TN2M0R0-stages. Their incidence of local recurrence as first relapse was quite similar (11/47:8/39). The related 4-year recurrence-free survival (57%, 37%), indicating a plateau-like curve of long-term survivors. These promising results have to be confirmed by larger studies. We conclude that the indication for surgery as the first treatment step for SCLC should be the same as for the other non-small subtypes of lung cancers. PMID- 7492379 TI - Stress influence on development of hepatocellular tumors in transgenic mice overexpressing TGF alpha. AB - We investigated whether stress increases tumorigenesis in male transgenic mice that overexpress the gene encoding human transforming growth factor alpha (TGF alpha). At the age of 10-15 months, these mice begin to develop spontaneous hepatocellular carcinomas at high incidence. The male TGF alpha mice were housed with their siblings (non-stressful environment), housed in social isolation, or housed with aggressive non-siblings (stressful environment). Some animals in each group were exposed once a week to a second stressor (swim stress), beginning at the age of 7 months. Housing with aggressive non-siblings increased neoplastic growth in the male TGF alpha mice: the incidence and multiplicity of liver tumors, and tumor burden were higher in these animals than in the sibling-housed mice. Among the isolated TGF alpha mice, only the tumor burden was increased, when compared with the sibling-housed TGF alpha mice. Swim stress significantly increased the incidence of liver tumors and tumor burden in the sibling-housed TGF alpha mice. Plasma levels of 17 beta-estradiol (E2) that are elevated in the TGF alpha mice, were modestly but significantly higher in the non-sibling housed transgenic mice than in the sibling-housed. Natural killer (NK) cell activity, reduced in these mice, was not affected by housing environment. These data suggest that stress promotes the growth of hepatocellular tumors in the male TGF alpha mice. Whether estrogens are involved in mediating this association remains to be determined. PMID- 7492380 TI - Modification of radiation-induced chromosome damage and micronucleus induction in mouse bone marrow by misonidazole and hyperthermia. AB - The effect of misonidazole (MISO), local hyperthermia (HT) and their combination on radiation-induced chromosome damage and micronucleus (MN) induction was studied in mouse bone marrow cells. It was found that MISO treatment did not enhance the clastogenic effect of radiation, which indicates a lack of radiosensitization of bone marrow chromosomes. But post-irradiation HT increased the frequency of aberrant cells and MN. A combination of MISO and HT produced a significant increase in the frequency of radiation-induced aberrant cells and MN at all the radiation doses as compared to radiation alone. The percentage of aberrant cells as well as the percentage of MN showed a linear quadratic increase with radiation dose in all the treatment groups. At higher radiation doses, cells with > 1 MN increased quadratically with a pronounced increase in cells bearing > 2 MN and severely damaged cells (SDCs) at radiation doses above 3.0 Gy in the HT and MISO+HT treated groups. Our results indicate that though MISO itself may not have a radiosensitizing effect on mouse chromosomes, a combination of MISO with HT can enhance the radiation damage in normal bone marrow. PMID- 7492381 TI - Effect of granulocyte-macrophage colony-stimulating factor (GM-CSF) on hematologic toxicity induced by high-dose chemotherapy in patients with metastatic breast cancer. AB - Twenty patients with recurrent metastatic breast cancer treated with high-dose myelosuppressive antineoplastic drugs (cyclophosphamide 2.5 g/m2 or epirubicin 130 mg/m2, both every 3 weeks) as first or second line chemotherapy were randomized in a prospective study to GM-CSF 5 micrograms/kg per day (n = 11) or control (n = 9). Significant reduction in granulocyte nadir duration (2 days with GM-CSF vs. 7 days) and severity (0.4 x 10(9)/l with GM-CSF vs. 0.2 x 10(9)/l) was found. No difference in frequency of neutropenic fever or antibiotic use could be observed. Even though the patients treated with GM-CSF at random were more heavily pretreated with chemotherapy, there was a surprisingly higher response rate in these patients as compared to the control-arm, namely 64% vs. 28.5%. However, this difference was not statistically significant. No severe side effects were seen, but presumably due to GM-CSF one patient developed an allergic type 1 reaction and one patient a possible pericardial exudation. Both were fully reversible after cessation of the cytokine treatment. PMID- 7492382 TI - Serum lactate dehydrogenase isoenzyme 1. An early indicator of relapse in patients with testicular germ cell tumors. AB - The present study aimed to evaluate serum lactate dehydrogenase isoenzyme 1 (S-LD 1) as an indicator of relapse for patients with testicular germ cell tumors. Twenty-seven patients were investigated with repeated determinations of S-LD-1 from diagnosis to relapse; 9 had seminoma and 18 nonseminomatous tumors. Eleven of 21 had raised S-LD-1 at relapse (4 with seminoma). Seven of the 27 patients had a raised S-LD-1 for median 2 months (1.4-4.5 months) before relapse was detected. Thus, S-LD-1 is of use, complementary to clinical examinations, determinations of serum alpha fetoprotein and human chorionic gonadotropin, and CT scans, in monitoring patients with testicular germ cell tumors for relapse. PMID- 7492383 TI - Immunoelectrophoretic differentiation of alpha-fetoprotein in disorders with elevated serum alpha-fetoprotein levels or during pregnancy. AB - Our study aims to make differential diagnosis by immunoelectrophoresis for some common conditions with elevated levels of serum alpha-fetoprotein (AFP). One hundred and nine cases with elevated AFP levels were included in this study: yolk sac tumor (n = 8), hepatocellular carcinoma (n = 26), gastric cancer (n = 12), chronic hepatitis (n = 27) and normal pregnancy (n = 36). Lectin agarose gel electrophoresis, antibody-affinity blotting, and immunoreaction were used to identify the specific patterns of AFP in the respective conditions. The results showed that there were three possible bands: L1, L2 and L3. Yolk sac tumor produced a prominent L2 band and a light L3 band. Hepatocellular carcinoma produced a prominent L1 band and a light L3 band. Gastric cancer produced only an L1 band. Chronic hepatitis had a light L1 band and a pronounced L3 band. In pregnancy, the AFP pattern is similar to that of hepatocellular carcinoma. Immunoelectrophoresis is a useful method facilitating the differentiation of AFP origins. PMID- 7492384 TI - Mesna/ifosfamide, mitoxantrone, etoposide, bleomycin, vincristine, prednisone (MINE-BOP) combination chemotherapy in the treatment of refractory and relapsed non-Hodgkin's lymphoma. AB - Twenty-one consecutive patients with refractory or relapsed non-Hodgkin's lymphomas were treated with a novel combination chemotherapy (MINE-BOP), comprising myelosuppressive (ifosfamide, mitoxantrone, etoposide) and non myelosuppressive (bleomycin, vincristine and prednisone) drugs. Median age of the patients was 42 years and all had intermediate or high-grade lymphoma. Fifteen patients had refractory disease. All patients had previously been treated with one or two regimens, containing anthracyclines. In all cases the duration between the last chemotherapy and the MINE-BOP regimen was shorter than 12 months. Response rate was 57% with 33% complete remission (CR). Median disease-free and overall survivals were 7 and 10 months respectively. The serum LDH level was the only significant prognostic factor in this study. The toxicity of this regimen was moderate with 24% of febrile neutropenia and 9% of microscopic hematuria. Toxic death due to febrile neutropenia was observed in one patient who had bone marrow involvement. To conclude, the addition of non-myelosuppressive drugs to the chemotherapy regimen and shortening the interval between the application of cytotoxic drugs as used in the present study did not show any improvement of response and survival in this group of patients. PMID- 7492385 TI - Chemotherapy and radiotherapy in locally advanced cervical cancer. AB - Radiotherapy has been standard therapy for locally advanced squamous cell cervical cancer. Neoadjuvant chemotherapy is being studied to improve responses and survival. We report a phase II study in locally advanced squamous cell cervical cancer (FIGO stages III and IVA) using chemotherapy with bleomycin, methotrexate and cisplatin (BMP) followed by radical radiotherapy. Of the 35 patients, 31 in stage III and 4 in stage IVA, 3 complete responses (CR) and 22 partial responses (PR) were achieved after chemotherapy treatment. Thirty-one patients completed radiotherapy; 19 achieved CR and 4 PR. Five-year actuarial survival for the entire group was 45% (95% confidence interval, 37-53%) with a median survival of 56 months. Patients with CR had a significantly better survival: the 5-year actuarial survival was 74% (95% CI, 59-89%). Recurrence developed in 4 of 19 patients. The most frequent side-effects were nausea and vomiting. Myelosuppression and impaired renal function also occurred. There was no evidence of radiotherapy toxicity enhancement. The stage and Karnofsky index were significant prognostic factors. It is concluded that BMP chemotherapy in advanced cervical cancer is effective and, followed by radiotherapy, allows a good control of this tumor. The group of patients with complete response have a low rate of recurrences and a long survival chance. PMID- 7492386 TI - Transrectal ultrasonically-guided core biopsies in the assessment of local cure of prostatic cancer after radical external beam radiotherapy. AB - Fifty-five patients were included in an extended follow-up after radical radiation therapy (RRT) for localized prostatic cancer (T1-3, Nx, M0). Local cure was assessed by a combination of digital rectal examination (DRE), transrectal ultrasound (TRUS) and systematic 'mapping' with TRUS-guided core biopsies (TGCB). After a mean follow-up of 6.8 years, 33% (18/55) of the patients were locally free of tumour, while in 67% (37/55) of cases residual cancer was demonstrated in the biopsies. Endocrine treatment did not influence the local cure rate, nor did the T stage of tumour grade at diagnosis or the cumulative radiation effect (CRE) values within the range of the present study. The sensitivity of DRE and TRUS was low; 37% and 20% respectively, while the specificity of the DRE and TRUS methods was 83% and 94% respectively. The conclusion of the study is that residual tumour was found in the high proportion of biopsied patients nearly 7 years after RRT and that multiple, TRUS-guided core biopsies are mandatory in the assessment of local cure in patients irradiated for prostatic cancer; both DRE and TRUS on their own are less reliable. PMID- 7492387 TI - External beam radiotherapy of localized prostatic adenocarcinoma. Evaluation of conformal therapy, field number and target margins. AB - The purpose of the present study was to identify factors of importance in the planning of external beam radiotherapy of prostatic adenocarcinoma. Seven patients with urogenital cancers were planned for external radiotherapy of the prostate. Four different techniques were used, viz. a 4-field box technique and four-, five- or six-field conformal therapy set-ups combined with three different margins (1-3 cm). The evaluations were based on the doses delivered to the rectum and the urinary bladder. A normal tissue complication probability (NTCP) was calculated for each plan using Lyman's dose volume reduction method. The most important factors that resulted in a decrease of the dose delivered to the rectum and the bladder were the use of conformal therapy and smaller margins. Conformal therapy seemed more important for the dose distribution in the urinary bladder. Five- and six-field set-ups were not significantly better than those with four fields. NTCP calculations were in accordance with the evaluation of the dose volume histograms. To conclude, four-field conformal therapy utilizing reduced margins improves the dose distribution to the rectum and the urinary bladder in the radiotherapy of prostatic adenocarcinoma. PMID- 7492388 TI - Field displacement during external radiotherapy in prostatic adenocarcinoma treated with radioactive 198Au implants and external irradiation. AB - The purpose of this work was to study displacement error and internal movements of the prostate during external beam radiotherapy. Verification films in the frontal (n = 194) and lateral (n = 64) portals were investigated in 14 patients treated with radioactive 198Au implants. Displacement errors of two implants were investigated. In seven patients, filling of the rectum and the bladder with contrast medium or isotonic saline was performed during CT investigation for planning purposes to detect movements of the prostate. Most (95%) of the displacement errors were less than 10 mm in the frontal portal and less than 15 mm in the lateral portals. No correlation to the patient's weight was found. The displacement errors were randomly distributed. The spatial relations between the implants were not altered during the treatments. Small movements of the prostate were observed. To conclude, the positioning system employed at present (laser) can be sufficient for the margins used (2 cm). In lateral portals, however, the system did not have the ability to detect a possible systematic displacement error from simulator to accelerator. The intention is to decrease the margins to 1 cm, which will necessitate a better positioning system. PMID- 7492389 TI - The impact of lymph node metastases on the results of treatment by primary radiotherapy and secondary surgery in oropharyngeal cancer. AB - The prognostic significance of lymph node metastases was evaluated in an unselected material of 427 oropharyngeal cancer patients treated primarily with radiotherapy. At first referral, palpable lymph node metastases were present in 60% of the patients. After irradiation, 60% of all palpable nodes had disappeared. As many as 50% of the nodes initially palpable in N3 patients vanished after irradiation. Recurrences were significantly increased in patients primarily with lymph node metastases (N1-3) compared with N0 patients; recurrences in the primary tumour site were (56% vs. 43%) in neck lymph nodes (40% vs. 9%) and in distant metastases (14% vs. 5%). Surgery was performed in 98 of 256 patients (38%) with recurrent or metastatic disease. The 5-year disease free survival rate after radiotherapy for N0 patients was an improvement (44%) on that of N1-3 patients (23%). The N-stage is an important prognostic factor for oropharyngeal cancer patients. PMID- 7492390 TI - Breast angiosarcoma after radiation therapy. PMID- 7492391 TI - Acute renal insufficiency due to viral infection caused by cytomegalovirus (CMV) during treatment with methotrexate. PMID- 7492392 TI - Aggressive hepatic leiomyosarcoma--an immunohistochemical evaluation of malignant potential using monoclonal antibody MIB-1. PMID- 7492393 TI - 5-Fluorouracil/levamisole induced intrahepatic fat infiltration imitating liver metastasis. PMID- 7492394 TI - Primary fibrosarcoma of the right ventricle--a case report. PMID- 7492396 TI - Tumor cell DNA content and radiation response in head and neck squamous cell carcinoma. PMID- 7492395 TI - Combined interferon and vinblastine treatment in advanced renal cell cancer. PMID- 7492397 TI - Systemic mitomycin C as second-line treatment for metastatic urothelial cancer. PMID- 7492398 TI - [Experience of the exclusive use of arteries in myocardial revascularization]. AB - OBJECTIVES: To present the initial experience of a surgical team from Santa Marta Hospital using exclusive arterial grafts on coronary bypass surgery. SETTING: Cardiology and Cardiothoracic Departments. POPULATION AND METHODS: From November 1987 to May 1994, 176 consecutive patients were submitted to surgical myocardial revascularization with the exclusive use of arterial grafts. The patients were operated by one cardiothoracic surgical team. A revision of clinical and angiographic characteristics of the patients was done along with the surgical procedure and intrahospital morbidity. After hospital discharge, follow-up was done by direct evaluation and/or by a questionnaire made using phone or letter on a periodical basis. RESULTS: The medium number of coronary bypasses performed were 2.07 +/- 0.8. The left internal mammary artery was the vessel more frequently used, specially to the myocardial area supplied by the left coronary descending artery. The right internal mammary artery was the second arterial graft more currently used. Gastroepiploic artery was utilized preferentially to coronary arteries supplying the inferior wall of the heart. Hospital mortality was 3.4%-6 patients. One hundred twenty patients have a normal life, without physical restrictions, 25% of the patients have symptoms of angina that is severe in 9. Mean follow-up time was 34 +/- 10 months. IN CONCLUSION: Coronary artery bypass surgery with exclusive use of arterial grafts can be executed with an acceptable mortality and low morbidity rates, even in urgent cases, with relieve of cardiac symptoms and good quality of life. PMID- 7492399 TI - [The pocket infection-erosion of permanent pacemakers: the results of a conservative approach without substitution of the components]. AB - OBJECTIVE: To assess the value of a morphologic classification of pocket lesions in eroded-infected permanent pulse generators and its influence in the efficacy or surgical management without removal and with system reutilization. DESIGN: Retrospective evaluation of patients, presenting with eroded-infected pacemaker, submitted to surgery. SETTING: Pacing Center and Cardiology Department of Faro District Hospital. PATIENTS: 43 consecutive patients treated by the same surgeon between 1985 and 1994. INTERVENTIONS: We considered three groups, based on morphologic characterization of the pocket lesions: G1-partial extrusion from the pulse generator with focal, adherent, discoloured and eroded skin; G2-identical to the previous group but with inclusion of lead segments or only lead extrusion; G3-tender pacing sites with fluctuation and, usually, fistula formation without the skin lesions of the other groups. In every patients a new ipsilateral generator pocket has been created just aside from the previous one with reimplantation of the same pacemaker. Failure was defined as the need for explantation of the system, with a new pacemaker implantation on the contralateral site, after the first attempt in G3 and a second in the other two groups. MEASUREMENTS AND RESULTS: The erosion-infection rate was 6.9%. Recurrences appeared in, 8.3%, 36.3% and 75% for, respectively, G1, G2 and G3. The initial and final success rates of G1 were significantly different from G3 success rates (p, respectively, < 0.07 and < 0.001). At the end of our study, reutilization surgery was successful in 91.4% of G1 and G2 assembled patients compared with 25% of the G3 patients (p < 0.003). Mean hospital stay for G3 patients was 7.4 +/- 1.0 days while G1 and G2 were managed as outpatients. The mean follow-up of patients without recurrences of pocket erosion-infection was 32.8 months. CONCLUSIONS: Our data suggest that a morphologic classification of lesions is very useful for treatment choice. It is possible, then, to select patients that may successfully managed by ipsilateral reimplantation without removal of pacing systems or hospital stay. The differences in the appearance of lesions and management results must reflect different etiopathogenic mechanisms. PMID- 7492400 TI - [The catheter ablation of atrioventricular nodal reentry tachycardia: the results of a mixed electrophysiological/anatomical technic]. AB - OBJECTIVE: To describe the results of radiofrequency catheter ablation of atrioventricular nodal reentrant tachycardia (AVNRT) by using a mixed electrophysiologic/anatomic mapping technique. DESIGN: Consecutive patients with AVNRT, submitted to AV node modification between November 1992 and March 1995. SETTING: Cardiology Department at an University Hospital. INTERVENTIONS: Twenty consecutive patients with AVNRT were submitted to AV node modification. The ablation technique included two sequential steps. Initially, a detailed electrophysiologic mapping was performed with the ablation catheter, positioned near the coronary sinus ostium, looking for a specific recording: fractionated atrial electrogram, suggestive "slow pathway" potential and a ratio of atrial: ventricular electrogram amplitude > 1. In case of failure, after 5 applications of radiofrequency energy, an anatomic technique was used. The fluoroscopic image, in left anterior oblique projection, was used to guide catheter progression, and the radiofrequency energy applied sequentially in the posterior (P), followed by medium (M) and anterior (A) septal areas if needed. Radiofrequency energy was applied a power of 16-36 watts for 30-60 sec. If a His bundle deflection > 0.0025 mV was recorded, energy was not applied. MEASUREMENTS AND RESULTS: Suppression of a AVNRT was initially obtained in 19 patients (95%). A mean of 8.3 +/- 6.1 energy application were required. Mean during of the entire procedure was 142 +/- 45 min and the fluoroscopy duration was 22 +/- 12 min. There were no complications. The location of successful ablation areas was: P in 15 patients, M in three and A in one. After a mean follow up of 10 +/- 6 months, two patients had recurrence of AVNRT. A second procedure was successful in the initially failed patient and in these two recurrences. CONCLUSIONS: A mixed electrophysiologic/anatomic mapping technique to perform radiofrequency catheter ablation of AVNRT was associated with high efficacy and no complications. PMID- 7492401 TI - [ST segment depression during exertion as a marker of left ventricular dysfunction in patients with aortic insufficiency]. PMID- 7492403 TI - [Intradialysis arterial hypotension in chronic hemodialysis and its relation to autonomic dysfunction]. PMID- 7492402 TI - [Homocysteinemia as a risk factor for myocardial infarct--the importance of age and of the homocysteinemia levels]. PMID- 7492404 TI - [The correlation between the severity of chronic mitral regurgitation evaluated by color-coded echo-Doppler and by angiocardiography with the "v" wave index in pulmonary capillary pressure/left ventricular systolic pressure]. PMID- 7492405 TI - [An echocardiographic comparison of left ventricular function in HIV-virus carriers and infected patients in different stages of immunosuppression]. PMID- 7492406 TI - [Insulinemia and coronary disease: the predictive value compared with "classical" cardiovascular risk factors]. PMID- 7492407 TI - [An analysis of the prevalence of the Arg403Gln, Gly584Arg and Leu908Val mutations in the beta-myosin heavy chain in Portuguese patients who are carriers of familial hypertrophic cardiomyopathy]. PMID- 7492409 TI - [Chronic heart failure: diastolic dysfunction versus systolic dysfunction]. PMID- 7492411 TI - [The evolution of the transmitral and transaortic gradient in aortic coarctation]. PMID- 7492410 TI - [Infectious endocarditis. The current diagnostic problems]. PMID- 7492408 TI - [Haptoglobin phenotypes and the pressure reaction to exertion in normotensive subjects]. PMID- 7492412 TI - [Dilated myocardiopathy in children: the doubts persist in 1994]. PMID- 7492413 TI - [Hypertensive heart disease: the definition of subgroups using left ventricular mass, the shortening fraction and the E/A ratio as parameters]. PMID- 7492414 TI - [The distribution of the E/A ratio in a population with essential arterial hypertension]. PMID- 7492415 TI - [Essential arterial hypertension: the clinical and echocardiographic parameters in relation to Na-Li countertransport]. PMID- 7492416 TI - [The effect of systolic and diastolic pressure on the cardiac repercussion of arterial hypertension. A contribution to its classification]. PMID- 7492417 TI - [Left ventricular hypertrophy: its prevention with enalapril]. PMID- 7492418 TI - Research in general practice: who is calling the tune? PMID- 7492419 TI - Investigation in general practice of patients with suspected heart failure: how should the essential echocardiographic service be delivered? PMID- 7492420 TI - Factors that influence the patient centredness of a consultation. AB - BACKGROUND: Research suggests that patients are more satisfied with patient centred consultations than with doctor-centred consultations and that some health measures are positively influenced by this type of consultation. Research on the factors that influence the patient centredness of the consultation is sparse but suggests that women doctors are more patient centred than men doctors. AIM: A study was designed, first, to confirm or reject the suggestion that women general practitioners are more patient centred than men general practitioners and, secondly, to determine some of the factors that might influence the patient centredness of a general practitioner. METHOD: The audiotaped consultations of 41 general practitioners were analysed using an instrument designed to measure patient centredness. The general practitioners were selected from a group of volunteers to represent both sexes as well as a wide range of age and experience. RESULTS: The results of the analysis showed that women general practitioners, particularly trainers, had higher patient-centredness scores than men general practitioners. This behaviour is characterized by an increased frequency of open questions and greater attention to patient offers (anything of potential significance that a patient brings to the general practitioner consultation). Additionally, the combination of sexes in a consultation seemed to have an effect on the interaction, with woman general practitioner/female patient dyads (pairings) having the highest median patient-centredness score, woman general practitioner/male patient and man general practitioner/male patient dyads scoring the same as each other and man general practitioner/female patient dyads having the lowest median scores. CONCLUSION: The results suggest that women general practitioners, in this sample, were more patient centred than men general practitioners. The results also suggest that inherent inequalities exist, with female patients receiving a more patient-centred service from general practitioners of their own sex than from general practitioners of the opposite sex. Male patients, on the other hand, seemed to fare equally well, irrespective of the sex of their general practitioner. Training is required to address these inequalities and to improve the level of patient centredness in the consultation. PMID- 7492421 TI - Development of a questionnaire to measure patients' satisfaction with general practitioners' services. AB - BACKGROUND: It is now a requirement that patients' satisfaction with the services obtained from their general practitioner should be surveyed. AIM: The aim of the study was to produce a reliable and valid multidimensional patient satisfaction questionnaire that could be used in general practice. METHOD: Items were originally derived from patients' responses to open-ended questions. The resulting 148-item Likert-scale questionnaire was completed by 1193 patients. General satisfaction items were removed from the set, and responses to remaining items underwent factor analysis. Subscales were produced from items representing each factor. Reliability and validity of each subscale were examined. RESULTS: Five subscales with a total of 40 items resulted from the factor analysis: doctors, access, nurses, appointments and facilities. Each subscale was internally reliable (Cronbach's alpha coefficient between 0.73 and 0.95), and initial tests of validity suggested that all subscales were valid. CONCLUSION: The study has resulted in a 40-item scale that has been found to be reliable and valid after initial tests. Further work to test the reliability and validity of the final version of the patient satisfaction questionnaire is described. PMID- 7492422 TI - Annual night visiting rates in 129 general practices in one family health services authority: association with patient and general practice characteristics. AB - BACKGROUND: Rates of night visiting by general practitioners have increased steadily over the last 30 years and vary widely between general practices. AIM: An ecological study was carried out to examine night visiting rates by general practices in one family health services authority, and to determine the extent to which differences in night visiting rates between practices could be explained by patient and practice characteristics. METHOD: The study examined the variation in annual night visiting rates, based on night visit fees claimed between April 1993 and March 1994, among 129 general practices in Merton, Sutton and Wandsworth Family Health Services Authority, London. RESULTS: Practices' annual night visiting rates varied from three per 1000 to 75 per 1000 patients. The percentages of the practice population aged under five years and aged five to 14 years were both positively correlated with night visiting rates (r = 0.38 and r = 0.35, respectively), as were variables associated with social deprivation such as the estimated percentage of the practice population living in one-parent households (r = 0.24) and in households where the head of household was classified as unskilled (r = 0.20). The percentage of the practice population reporting chronic illness was also positively associated with night visiting rates (r = 0.26). The percentages of the practice population aged 35 to 44 years and 45 to 54 years were both negatively associated with night visiting rates (r = -0.34 and r = -0.31, respectively) as was the estimated list inflation for a practice (r = -0.31). There was no significant correlation between night visiting rates and the distance of the main practice surgery from the nearest hospital accident and emergency department. There was also no association between night visiting rates and permission to use a deputizing service. In a stepwise multiple regression model, the multiple correlation coefficient was 0.56 with four factors (percentage of the practice population aged under five years, percentage aged 35 44 years, percentage who were chronically ill and estimated list inflation) explaining 32% of the variation in night visiting rates. CONCLUSION: Only about one third of the variation in night visiting rates between practices could be explained by patient and practice variables derived from routine data. Population based research using data collected on individual patients and practices is required to improve current understanding of the patient and practice characteristics that influence the demand for night visits and of why night visiting rates vary so widely between practices. PMID- 7492423 TI - Assessment of the completeness and accuracy of computer medical records in four practices committed to recording data on computer. AB - BACKGROUND: General practice computer databases are being increasingly seen as a source of data for public health monitoring and commissioning. Such ambitions depend on routine clinical data being recorded with acceptable completeness and accuracy. AIM: The aim of this study was to assess the completeness and accuracy of the computer medical records in four high-recording general practices. METHOD: Four general practices in the Trent Region that use the EMIS computer system, and were known to be high recorders of clinical data on their computer databases, were selected. A retrospective analysis of the computer records, a prospective comparison of a sample of computer records with manual records, and a prospective comparison between videorecorded consultations and their manual and computer records were undertaken. RESULTS: Checks for completeness in computer recording of diabetes mellitus and glaucoma showed high levels of accurate recording, 97% and 92% respectively. Prevalence rates between practices were reasonably comparable. No practice consistently, across 10 diagnoses, recorded prevalences higher or lower than the other practices; those diagnoses with recognized objective diagnostic criteria were recorded with a more consistent prevalence than those without. Lifestyle data recording was low; overall, smoking habits and alcohol consumption were recorded for 52% and 38% of patients aged over 16 years, respectively. Comparison of the manual records with the computer records showed that the computer records were sufficiently complete with regard to diagnoses (82% of all items recorded), prescriptions (100%) and referrals (67%), but missed most of the remaining data that a manual record captured. The videorecorded validation study showed that there were no important lapses in the recording of diagnoses, prescriptions or referrals when the computer recording was compared to the actual process of the consultations. CONCLUSION: In these four high-recording practices the data in computer records were of sufficient completeness and accuracy to allow meaningful data aggregation for some diagnoses, prescriptions and referrals. Standardized protocols for defining which patients are included and excluded from major disease groups are required. PMID- 7492424 TI - Ebeltoft project: baseline data from a five-year randomized, controlled, prospective health promotion study in a Danish population. AB - BACKGROUND: There is increasing political pressure on the medical profession to approach welfare diseases, such as coronary heart disease and diabetes, through prevention. General practitioners are required to offer regular health checks to healthy people, in spite of the lack of scientific evidence for the universal need, usefulness and side effects of such an intervention. Randomized controlled trials are needed. AIM: A study was carried out to investigate people's interest in participating in health checks and in discussions about health with their own general practitioner, participants' health status, the proportion who received health advice following health checks, and the lifestyle goals they set following discussion with their general practitioner. This study reports the baseline data from a five-year randomized, controlled, prospective, population-based study in general practices in Ebeltoft, Denmark. METHOD: All general practitioners from the four practices in Ebeltoft and a random sample of 2000 people aged between 30 and 50 years were invited to participate. Participants were randomly divided into three groups--one control group and two intervention groups. One intervention group were given a health check which included being screened for cardiovascular risk factors, lung and liver function, fitness, sight and hearing and an optional test for the human immunodeficiency virus (HIV); this group received written feedback from the general practitioner. The other intervention group were also given a health check and written feedback; in addition, they were given the opportunity to attend their general practitioner to discuss preventive health. RESULTS: A total of 1370 people participated in the study (69% response rate). Health advice was given to 76% of 905 participants following health checks. Almost all of the 456 participants (96%) who were offered the opportunity of discussing their health with their general practitioner took up the offer; 64% of the 456 participants reported that they had decided to undertake lifestyle changes. Eleven of those who discussed their health with the doctor were referred to a specialist (2%). CONCLUSION: There was considerable interest in participating in health promotion. Three out of four of those having a health check were given health advice. Two out of three of those offered a health talk with the general practitioner appeared willing to make relevant lifestyle changes. Long-term follow up is needed to determine effects and side effects of health checks and health talks. PMID- 7492425 TI - Implementation of quality assurance and medical audit: general practitioners' perceived obstacles and requirements. AB - BACKGROUND: The introduction of quality assurance and medical audit has been an important development in general practice. However, the introduction of such programmes does not necessarily mean they are implemented by general practitioners. AIM: A study was undertaken to describe the problems and requirements perceived by general practitioners in relation to the implementation of quality assurance and medical audit in general practice. METHOD: Interviews were carried out with a stratified sample of 120 Dutch general practitioners. Knowledge, acceptance and application of quality assurance and medical audit activities were investigated, and perceived problems and requirements in implementing quality assurance and audit activities were explored. RESULTS: General practitioners in the Netherlands were generally positive towards quality assurance activities, but had little experience of carrying out such activities. The most frequently mentioned obstacles to implementing quality assurance activities concerned lack of time, colleagues' negative attitudes and fear of assessment and criticism by colleagues. Requirements for implementing quality assurance included having regular meetings with colleagues about quality assurance, having information on the aims and methods of quality assurance, having data from other practices with which to compare performance, having support in data collection, in audit in the practice and in setting up local peer review, and having financial support. The most important factor predicting the actual application of quality assurance activities was found to be knowledge of specific quality assurance activities. CONCLUSION: Well-designed programmes for the implementation of quality assurance and medical audit, using a variety of different interventions, have to be developed. Such programmes should include the training of professionals in the concepts and methods of quality assurance as well as the provision of financial support for quality assurance activities. PMID- 7492426 TI - Increasing the number of drugs available over the counter: arguments for and against. AB - Many drugs previously restricted to prescription only status are being reclassified as pharmacy only status and hence are becoming available over the counter to patients. A general practitioner should make enquiries about a patient's self-medication practices before deciding on treatment for the patient. Over-the-counter medicines are considered safe and their increased use indicates that patients are taking greater responsibility for their own health and possibly taking some of the financial burden of drug treatment from the National Health Service. The retention of their restriction to pharmacy only sale provides some additional protection for patients and promotes the role of pharmacists in the care of patients. However, having more drugs available for self-treatment may encourage patients to believe that there is a drug treatment for every ailment. Increasing the range of drugs available over the counter increases the risks of interactions and adverse reactions and of self-treatment being undertaken when medical aid should have been sought. For general practitioners to recommend positively use of over-the-counter preparations may involve some medicolegal risks, and the potential savings to the NHS may prove illusory. Education for patients and better communication between general practitioners and community pharmacists are required to allow easier availability of modern medicines to patients in order to bring the benefits anticipated. PMID- 7492427 TI - Clinical practice and medical research: bridging the divide between the two cultures. AB - The failure of the results of many research studies to be integrated into everyday clinical practice is both well documented and much decried. In the writings on why medical research and clinical practice have remained separate cultures, two issues have not been sufficiently debated. First, are medical researchers addressing the problems that cause clinicians the most concern in their consultations with patients, and secondly, are the results of research studies being presented in a manner that clinicians can both understand and use? This discussion paper highlights primary care clinicians' urgent need for information on the predictive value of the symptoms and signs seen in everyday clinical practice. Medical research has still to provide this information, often leaving general practitioners with inadequate predictive information on which to make early diagnoses, for example, on whether a patient with chest pain has a pulmonary embolus, or a child with pyrexia and rash has meningococcal septicaemia. The format in which research information is commonly presented is discussed; it has been shown that epidemiological terms used in studies are impenetrable to most clinicians. Additional ways of framing research information need to be devised that present such research information in a narrative format and numerical format, emphasizing the effects of management decisions as well as diagnostic categories, and for use in individual consultations as well as describing populations. Only then will clinicians be able to integrate into their everyday clinical practice the potentially valuable information provided by medical research. PMID- 7492428 TI - Headache: an ophthalmological problem. PMID- 7492429 TI - Hormone replacement therapy. PMID- 7492430 TI - Haemophilus influenzae vaccine: maximizing uptake. PMID- 7492431 TI - Warfarin in stroke prevention. PMID- 7492433 TI - Fourth national morbidity study. PMID- 7492432 TI - Aspirin and acute myocardial infarction. PMID- 7492434 TI - Acutely infected Langerhans cells are more efficient than T cells in disseminating HIV type 1 to activated T cells following a short cell-cell contact. AB - Most human immunodeficiency virus type 1 (HIV-1) infections involve sexual contact and virus passage across mucosal surfaces. While Langerhans cells (LCs) and dendritic cells (DCs) have been implicated in mucosal infection, their role is undefined. Here we demonstrate that acutely HIV-1-infected LCs and DCs effectively transmit virus to uninfected, activated T cells. Cocultivation of these cells results in massive virus production that requires a short cell-cell contact; as little as 30 min contact time is sufficient for HIV-1-pulsed DCs to infect their target T cells. Furthermore, surface-bound virus inactivation by trypsin does not significantly decrease the efficiency of virus transmission by LC/DCs, suggesting rapid internalization of virus. This effective virus transfer by infected LCs and blood-derived DCs requires prior activation of T cells. Surprisingly, cocultivation of acutely infected T cells with uninfected, activated target T cells results only in low virus production, even with T cell tropic virus. We conclude that LCs and DCs are not only important targets of HIV 1 infection, but may also play a key role in the early dissemination of virus to T cells they encounter in skin or lymphoid tissue. PMID- 7492435 TI - Costimulation of CD4+ T cells via CD28 modulates human immunodeficiency virus type 1 infection and replication in vitro. AB - Stimulation of human immunodeficiency virus type 1 (HIV-1)-infected donor peripheral blood mononuclear cells (PBMCs) via the TCR-CD3 complex induces HIV-1 production in vitro (Zarling JM, et al.: Nature [London] 1990;347:92; Haffar OK, et al.: J Virol 1992;66:4279; Moran PM, et al.: AIDS Res Hum Retroviruses 1993;9:455). However, in addition to the primary stimulatory signal delivered through the TCR-CD3 complex, optimal T cell activation requires secondary or costimulatory signals delivered via various T cell accessory proteins (Alton A, et al.: Adv Immunol 1990;48:227). In this article we explore the role of costimulation of T cells via CD28 in HIV-1 replication. Ligation of CD28 with either a CD28-specific MAb or by coculture of PBMCs with Chinese hamster ovary (CHO) cell lines stably expressing either of the CD28 counterreceptors, B7-1 (CD80) or B7-2 (CD86), concomitant with stimulation via CD3, results in increased virus replication compared to stimulation via CD3 alone. CD28 ligation also augments de novo infection of CD3-stimulated seronegative donor PBMCs with cell free virus. Increased virus replication following CD28 ligation is not solely attributed to increased levels of endogenous IL-2, because addition of an anti-IL 2-neutralizing antibody only partially inhibits the response. In contrast, interfering with the interaction between CD28 and its counterreceptors on antigen presenting cells (APCs) using CTLA4Ig effectively inhibits virus replication. At high concentrations CTLA4Ig also reduces cell proliferation. These in vitro results suggest that CD28 plays a central role in HIV-1 replication and that interfering with the CD28 costimulatory pathway may modify the course of HIV-1 infection. PMID- 7492436 TI - Transmembrane P-glycoprotein (P-gp/P-170) in HIV infection: analysis of lymphocyte surface expression and drug-unrelated function. AB - P-glycoprotein (P-gp/P-170), a transmembrane efflux pump known to be one of the mechanisms responsible for multidrug resistance in cancer therapy, is constitutively expressed in several solid human tissues as well as in normal peripheral blood lymphocytes and bone marrow cells. In particular, this molecule has been associated with the transport of perforin and other cytolysins in natural killer (NK) and T cytotoxic lymphocytes. In the present study, we analyzed peripheral blood lymphocytes (PBLs) from controls and HIV+ patients for phenotypic expression and function of the P-gp/P-170 molecule. We found that 90% of all PBL subsets (i.e., CD4+, CD8+, CD56+, and CD19+ cells) expressed surface P gp/P-170 both in controls and HIV+ patients. However, a significant decrease in CD4+/P-170+ and CD19+/P-170+ cells was observed in HIV+ individuals with respect to controls. PHA and IL-2 stimulation of PBLs was unable to increase the expression of P-gp/P-170 both in controls and HIV+ patients, despite the increased detection of the CD25 molecule. On the other hand, stimulation with anti-CD3 determined a significant increase in lymphocyte P-gp/P-170. The function of P-gp/P-170, assessed by a flow cytometric assay for rhodamine-123 (Rh123) efflux, was significantly reduced in CD16+ NK cells and CD19+ B cells from HIV+ patients. The Rh123 efflux by NK cells correlated (p < 0.01) with the NK cytotoxicity against the 51Cr-labeled K562 cell line. Last, the effect of the antiretroviral drugs AZT, ddI, and ddC on P-gp expression and function was evaluated. The dideoxynucleoside compounds did not inhibit P-gp/P-170 function of normal mononuclear cells in vitro, and did not increase P-gp/P-170 expression in vivo, in patients undergoing antiretroviral therapy with AZT. These findings provide further evidence of a possible involvement of the P-gp/P-170 system in specific immunological lymphocyte functions, and especially in cytotoxic-type functions. In addition, it is possible to suggest, on the basis of our experimental data, that the dideoxynucleoside class of antiretroviral agents does not contribute to the phenotypic and functional alterations related to P glycoprotein during HIV infection. PMID- 7492437 TI - Gag-specific cytotoxic responses to HIV type 1 are associated with a decreased risk of progression to AIDS-related complex or AIDS. AB - The duration of human immunodeficiency virus (HIV-1) infection prior to the development of AIDS is variable, and for most patients the exact time of infection is not known. A group of 38 HIV-1-infected subjects was tested while asymptomatic for comparative cytotoxic lymphocyte responses to the Gag and envelope antigens of HIV-1. Twenty of the 38 patients had no detectable primary cytotoxic T lymphocyte (CTL) response to Gag, and this was associated with a relative risk of 1.89 for progression to ARC or AIDS during the subsequent 3 to 40 months of observation when compared with patients who had Gag-specific CTL activity at the beginning of the observation period. In contrast, no significant association was observed between envelope-specific cytotoxic activity and disease progression. Other patient characteristics, including CD4+ T lymphocyte counts and antibody levels to the p24gag protein, measured at the start of observation, did not correlate with disease progression during the observation period. This suggests that the anti-Gag CTL response may be protective during HIV-1 infection. PMID- 7492439 TI - Cytokine-containing liposomes as adjuvants for HIV subunit vaccines. AB - Dehydration-rehydration liposome vesicles (DRVs) containing various cytokines were evaluated for their ability to induce delayed-type hypersensitivity (DTH) and humoral immunity to the recombinant envelope protein rgp120 of the MN strain of human immunodeficiency virus type 1 (HIV-1). The DRVs trapped approximately 25% of the radiolabeled cytokines and approximately 17% of the radiolabeled rgp120 that were added. The level of trapping was greater than the aqueous volume of the DRVs, indicating association of the proteins with the lipid bilayer. Flow cytometric analysis using antibody to rgp120 or the V3 loop of rgp120 showed the diameter of the DRVs to be 2-7.5 microns. Transmission electron microscopy confirmed the heterogeneity in size of the DRVs and revealed morphological heterogeneity. Transmission electron microscopy with immunogold labeling also revealed the presence of rgp120 on the surface of the DRVs. In vitro bioassays demonstrated slow leakage of biologically active cytokines from DRVs soaked in tissue culture medium containing serum. Mice injected subcutaneously three times at 14-day intervals with DRVs containing 15 micrograms of rgp120 plus interleukin 6 (IL-6) or interferon gamma (IFN-gamma) produced significantly greater DTH responses than mice injected with DRVs containing rgp120 alone. Soluble rgp120 plus soluble IFN-gamma produced DTH in some experiments, but of lower magnitude than the comparable DRVs. Interleukin 6, but not IFN-gamma, increased the antibody titer to rgp120 when included in the DRVs. The mice did not develop antibodies to IFN-gamma or IL-6. Induction of DTH by vaccines may increase protection from viral pathogens such as HIV. Cytokine-containing liposomes may be an effective adjuvant for the induction of a DTH response to envelope-antigen subunit vaccines. PMID- 7492438 TI - Highly attenuated HIV type 2 recombinant poxviruses, but not HIV-2 recombinant Salmonella vaccines, induce long-lasting protection in rhesus macaques. AB - Immunization schemes employing priming with vector-based vaccine candidates followed by subunit booster administrations have been explored and shown to have merit in the human immunodeficiency virus type 1 (HIV-1) and simian immunodeficiency virus systems. In this study, we have assessed the priming capacity of highly attenuated poxvirus vector (NYVAC and ALVAC)-based HIV-2 recombinants, as well as Salmonella typhimurium HIV-2 recombinants in rhesus macaques. ALVAC- and NYVAC-based vaccine candidates expressing the HIV-2 gag, pol, and env genes or NYVAC-based recombinants expressing either gp160 or gp120 were used to immunize rhesus macaques in combination protocols with alum adjuvanted HIV-2 rgp160. Following intravenous challenge exposure with 100 infectious doses of the HIV-2SBL6669 parental virus genotype mixture, seven of eight animals were protected from infection. The seven protected animals were rechallenged 6 months postprimary challenge, without additional booster inoculations, with the same dose of the HIV-2SBL6669 parental virus. Five of the seven animals remained protected against HIV-2 infection at 6 months following the second challenge. In contrast, oral immunization with recombinant Salmonella expressing the HIV-2 gag and the gp120 portion of the envelope either alone or in combination with alum-adjuvanted rgp160 failed to confer protection. These results suggest that the NYVAC- and ALVAC-based recombinants may confer long lasting protection and that these two highly attenuated poxvirus vaccine vectors may represent promising candidates for developing an acquired immunodeficiency syndrome vaccine. PMID- 7492440 TI - Induction of potent humoral and cell-mediated immune responses following direct injection of DNA encoding the HIV type 1 env and rev gene products. AB - DNA vaccines have the potential of giving rise to a potent cell-mediated immune response by inducing intracellular synthesis and subsequent antigenic presentation of encoded antigens. We have tested a DNA vaccine specific for human immunodeficiency virus type 1 (HIV-1) by the injection of animals with expression plasmids encoding the HIV-1 envelope protein and the Rev regulatory protein. Injection of both plasmids into mice, rabbits, or macaques was found to induce high levels of specific antibodies capable of efficiently inhibiting both HIV-1 infection and envelope-mediated cell fusion. A readily detectable delayed-type hypersensitivity (DTH) response was demonstrable in injected mice and lymphocytes derived from these proliferated in response to an HIV-1 envelope V3 loop-specific peptide. Interestingly, the injected mice or macaques also developed a strong cytotoxic T lymphocyte (CTL) response against target cells pulsed with the V3 peptide. Taken together, these data demonstrate that injection of HIV-1 gene expression plasmids can induce potent humoral and cell-mediated immune responses and suggest that DNA vaccines may prove to be significantly beneficial as a means of immunizing against HIV-1. PMID- 7492441 TI - Expression and purification of the HIV type 1 Rev protein produced in Escherichia coli and its use in the generation of monoclonal antibodies. AB - We have developed a simple and rapid procedure for the purification of large amounts of Rev protein overexpressed in E. coli. The purification method, which does not require denaturation of the protein, takes advantage of the positively charged nature of Rev and the ability of Rev to interact with nucleic acids. The purified protein was used to develop three novel murine monoclonal antibodies against Rev. Using fusion proteins between glutathione S-transferase (GST) and various fragments of the Rev protein, we mapped the specificity of these antibodies to different regions of the Rev protein. One antibody, 3H6, is directed against the nucleolar localization/RRE-binding domain of Rev between amino acids 38 and 44. Another antibody, 3G4, recognizes an epitope between amino acids 90 and 116 of Rev. A third antibody, 2G2, does not recognize any of the fusion proteins, and may be directed against a conformational epitope. All three antibodies are able to detect Rev on Western blots and to immunoprecipitate Rev under native conditions. However, only 3H6 and 3G4 immunoprecipitate Rev under denaturing conditions and are able to detect Rev expressed in transfected cells by indirect immunofluorescence. These antibodies should prove useful in further studies of Rev function. PMID- 7492443 TI - Studies of complement-activating antibodies in the SIV/macaque model of acute primary infection and vaccine protection. AB - Questions regarding the potential impact of complement-activating antibodies on lentivirus pathogenesis and vaccine development were addressed in the SIV/macaque model by evaluating sera for activity related to complement-mediated, antibody dependent enhancement (C'-ADE) of SIV infection in vitro. C'-ADE activity in sera obtained during acute primary infection in macaques inoculated with SIVmac251 appeared before neutralizing antibodies and coincided with the initial peak and decline of plasma antigenemia. The power of C'-ADE activity (i.e., virus production measured by p24 immunoassay) decreased as titers of neutralizing antibodies increased in these animals, suggesting a balance in the net effect between C'-ADE and neutralizing activities in vitro. Antibodies with C'-ADE activity were also induced in macaques immunized with live-attenuated SIVmac239/nef-deletion or primed with recombinant SIVmne gp120 vaccinia virus and boosted with SIVmne rgp160. The titer (i.e., last serum dilution to show enhancement), peak (i.e., serum dilution producing the greatest enhancement as measured by p24 production), and power (i.e., magnitude of p24 production at the peak titer) of C'-ADE activity in sera obtained from vaccinated macaques on the day of challenge were comparable to those of sera from infected macaques and showed no correlation with vaccine outcome, where some protected animals had C' ADE profiles that resembled those of unprotected animals. The results of these studies suggest that antibodies having C'-ADE activity in vitro could contribute to virus replication or, alternatively, to virus clearance during the acute stage of SIV infection in macaques.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7492442 TI - Simian immunodeficiency virus infection of macaque primary placental cells. AB - We have characterized the ability of a simian immunodeficiency virus, SIVmne strain E11S, to infect macaque placental trophoblast and Hofbauer cells. These primary placental cells were permissive to SIVmne infection, regardless of gestational age. Virus production by the infected cells was determined as time dependent viral core antigen p27 production, followed by verification of the proviral gag/LTR DNA sequences in the infected cells using a polymerase chain reaction assay. Of more than six placentas tested, SIVmne infection of placental cells at an early gestational age (i.e., days 55 or 78) produced more than 10 fold the amount of virus core antigen p27 than did placental cells infected at a late gestational age (i.e., days 135 or 165). In addition, SIVmne infection of trophoblast cells was inhibited by SIVmac neutralizing macaque serum but not by normal serum, indicating the specificity of virus infection. Furthermore, the amount of SIV core antigen p27 produced by the virus-infected trophoblast and Hofbauer cells was shown to be dependent on the multiplicity of virus infection. Collectively, our results indicate that macaque trophoblast and Hofbauer cells can be infected by SIV and that both gestational age and viral dose may play a role in the extent of viral infection. PMID- 7492444 TI - HIV glycoprotein 41 and complement factor H interact with each other and share functional as well as antigenic homology. AB - We have shown that complement factor H (CFH) interacts with HIV-1 at the level of the sequence Env 105-119, contained in the C1 domain of gp120. CFH interaction with HIV was evident only after dissociation of the Env complex induced by exposure to sCD4. We hypothesized that CFH could act as a gp41 analog in the interaction with Env 105-119. A panel of partially overlapping, synthetic peptides reproducing the extracellular portion of gp41 was therefore used to compete the binding of CFH to Env 105-119. Three sets of peptides that competed this interaction were identified. These peptides defined a region of functional homology between the gp41 molecule and CFH (Env 580-600), and two regions of interaction (Env 620-640 and Env 650-670). In addition to this, a monoclonal antibody directed against peptide Env 580-600 and a polyclonal mouse antiserum raised against recombinant gp41 were shown to recognize CFH in Western blots and ELISA, respectively, also defining a region of antigenic homology between gp41 and CFH. These data provide evidence for interaction and molecular mimicry between an HIV structural protein and a negative regulator of the complement pathway. We show here that CFH can interact with both HIV Env proteins, suggesting a possible and efficient mechanism of downregulation of the complement cascade at the surface of infected cells. PMID- 7492446 TI - Increased risk for a second retroviral infection (SIV or STLV type I) for wild African green monkeys already infected by one retrovirus in Senegal (West Africa). PMID- 7492445 TI - Molecular characterization of an HIV type 1 isolate from Burundi. PMID- 7492447 TI - A new HTLV type II subtype A isolate in an HIV type 1-infected prostitute from Cameroon, Central Africa. AB - Among 332 female sex workers in Douala, Cameroon, 113 were HIV-1 seropositive, 3 were HTLV-I seropositive, and only 1 had specific anti-HTLV-II antibodies. By cocultivation with BJAB cells, an HTLV-II was isolated from the peripheral blood mononuclear cells of this 32-year-old woman coinfected by HIV-1. This new African HTLV-II isolate (PH230PCAM) belongs to the molecular subtype A, exhibiting, however, a nucleotide variability of 2.4% and 0.8%, vis-a-vis the MO prototype, in the LTR and in the gp21 env gene, respectively. These data, as well as the previous findings of another HTLV-II subtype A in a Ghanean prostitute, suggest that this viral subtype had been imported into Africa, while the HTLV-II subtype B, described in remote areas of Zaire, Gabon, and Cameroon, could be a genuine African HTLV-II, present in this continent for a long period of time. PMID- 7492448 TI - Yet another subtype of HIV type 1? PMID- 7492449 TI - HIV type 1 sequence subtype G transmission from mother to infant: failure of variant sequence species to amplify in the Roche Amplicor Test. PMID- 7492452 TI - Moving managers into new leadership roles. PMID- 7492450 TI - 8th European Cancer Conference. Paris, France, October 29-November 2, 1995. Abstracts. PMID- 7492451 TI - Nursing leaders of the next century. PMID- 7492453 TI - Action learning and developing competent self-directed work teams. PMID- 7492454 TI - Sexual activity and heart disease. PMID- 7492455 TI - AIDS-related behaviour in adolescents: intervention programme with international application. AB - Adolescents and young adults compromise one of the fastest-growing categories of AIDS cases. The results of an opinion survey taken of 3242 adolescents raise serious doubts about the prevalent view that high-risk behaviour is attributable to insufficient information and education on AIDS: female adolescents are found to be at risk despite having the proper knowledge. Five chief factors affected the knowledge and behaviour of adolescents: age, gender, origin, family profession, and parental education. Each of the subpopulations (females, younger adolescents, children in rural areas) has its own characteristics and needs. We describe a permanent and long-term intervention programme, which might be applied in any country. PMID- 7492456 TI - Follow-up of patients with suspected venous thrombosis but negative radionuclide venography. AB - A retrospective review of the outcome of patients presenting to the district general hospital with suspected deep vein thrombosis (DVT) but normal radionuclide venogram. Of 387 scans, 141 (36%) were reported as positive, 81 (21%) as equivocal, and 165 (43%) as negative. The casenotes of 150 (91%) of those with negative scans were reviewed of which 124 were performed as part of the investigation of suspected DVT. During the next 3 months there were 13 DVTs and 7 with pulmonary emboli in this group, 4 of the latter proving fatal (confirmed at postmortem in 3). From a review of the initial clerking, the pre venogram likelihood of DVT was graded as low in 54 (43%), medium in 23 (18%), and high in 45 (36%). DVT or PE occurred in none of those with a low probability, 9% of those with medium, and 38% of those with high probability. The results suggest that radionuclide venography is insufficient to exclude DVT when suspected clinically. PMID- 7492457 TI - Imaging and staging of small-cell lung cancer: is there a future role for octreotide scintigraphy? AB - In small-cell lung cancer (SCLC), CT scan remains the most accurate imaging modality for evaluating local extension and specific sites of metastatic disease. The role of nuclear medicine in the work-up of SCLC is still limited to the detection of bone metastases. Recently, a new potential diagnostic tool has been introduced based on the presence of somatostatin receptors in SCLC. With the use of radiolabelled somatostatin analogues it is hoped that an equally effective but simpler staging system has been found that gives a better separation of prognostic subgroups. This article reviews the role of nuclear medicine in general and somatostatin receptor scintigraphy in particular in the imaging and staging of SCLC. Clinical value in terms of sensitivity and specificity is discussed in relation with other imaging and staging modalities. PMID- 7492458 TI - Effect of alcohol on body core temperature during cold-water immersion. AB - This study investigates the influence of alcohol on body core temperature during cold-water immersion in human volunteers. In this randomised, double-blind, placebo-controlled crossover trial, 8 healthy male volunteers were randomised to drink 1 litre of beverage containing 50 g alcohol or placebo before 20 degrees C cold-water immersion for 1 hour. Body core temperature was measured before cooling and after 1 hour of immersion with an infrared ear thermometer. After a 1 week interval subjects crossed to the other study branch. In the placebo test, mean temperature decreased after immersion by 0.66 degrees C; after the alcohol mean temperature decreased by 1.0 degrees C (mean difference 0.34, 95% CI 0.14 0.53, P = 0.002). PMID- 7492459 TI - Routine clinical biochemistry tests collected from patients dying in hospital. PMID- 7492460 TI - Auditory rehabilitation: needs and realities on long-stay wards for elderly people. AB - A survey was made of the problem of age-related hearing impairment within a sample of National Health Service long-stay wards for elderly people. Detailed assessment of hearing was completed for 188 residents derived from 15 long-stay wards, located within nine Scottish hospitals. The sample residents were more than four times as likely to need a hearing aid as their contemporaries in the general population. Of the 131 people considered to be potential hearing aid candidates, 103 (78%) demonstrated a positive attitude towards hearing aids ownership. Medical policies on the study wards were not considered sensitive to the needs of the hearing-impaired; screening was haphazard and specialist referral problematic. PMID- 7492461 TI - Hepatitis and HIV: interrelationship and interactions. AB - Hepatitis in the context of HIV infection is commonplace, but the causative agent is not always readily identifiable. This review highlights the differential diagnosis of hepatitis in that context, examining the interrelationships and interactions between the four main hepatitis viruses and HIV. PMID- 7492462 TI - Deep vein thrombosis. AB - Deep vein thrombosis (DVT) is a common condition. Most cases arise as complications during the perioperative period. This can largely be prevented by adequate prophylaxis, principally using low-dose subcutaneous heparin. Only a minority of DVTs produce serious complications, but it is not currently possible to predict the clinical behaviour of any DVT, once formed. For this reason, any identified DVT should be vigorously treated. The mainstay of treatment remains systemic anticoagulation with heparin and then warfarin. Warfarin should be continued for 1 month in postoperative cases and 3 months in spontaneous cases, provided there is no ongoing predisposing factor. Recurrent spontaneous DVT formation is an indication for lifelong anticoagulation. Recent evidence suggests that the subcutaneous route of administration of heparin has advantage over traditional intravenous infusion. Some large DVTs require thrombolysis, and it is now possible to treat the underlying anatomical defects with angioplasty and endovascular stenting, although the long-term outcome of these procedures has not yet been established. For patients with contraindications to the use of anticoagulants, a variety of (temporary and permanent) percutaneously inserted vena caval filters are now available. The principal complications of DVT are pulmonary embolism, which may be fatal, and the development of a postphlebitic leg. The avoidance of these depends on adequate prophylaxis and vigorous treatment of the primary DVT. PMID- 7492464 TI - Drugs affecting the leukotriene pathway in asthma. AB - Drugs affecting the leukotriene pathway are emerging as a new class of asthma treatment--the first for more than 20 years. Development of these leukotriene antagonists should lead to greater understanding of the underlying processes in asthma and may result in a safe, oral, anti-asthmatic drug to benefit particular groups of asthmatic patients. PMID- 7492463 TI - Digoxin in clinical practice: sorting out the facts. AB - The aim of this review is to sort out from the huge amount of facts related to digoxin in the literature those pertinent to current clinical practice. Seven aspects of digoxin are discussed: its mechanism of action, therapeutic controversy, clinical efficacy, avoidance of toxicity, manifestations of toxicity, indications, and its use in coronary artery disease. PMID- 7492465 TI - Viral hepatitis: an overview. PMID- 7492466 TI - Compound opioid/paracetamol analgesics: misuse and dependence. AB - Ten cases of individuals with a history of chronic compound opioid/paracetamol misuse are described. Potentially useful indicators of toxicity are suggested. PMID- 7492467 TI - C. albicans septic arthritis in a patient with AIDS. PMID- 7492468 TI - Munchausen's syndrome (HIV infection): a trap for the unwary. PMID- 7492469 TI - Paget's disease: ocular complications of disodium pamidronate treatment. AB - Three cases of uveitis associated with disodium pamidronate have been notified to the Committee for Safety of Medicines. Two new cases of bilateral anterior uveitis and episcleritis after administration of intravenous disodium pamidronate are described. PMID- 7492470 TI - Salmonellosis complicated by myocardial abscess and cerebral mycotic aneurysms. PMID- 7492471 TI - Massive endometrial polyp after tamoxifen therapy. PMID- 7492472 TI - Persistent mullerian duct syndrome. AB - Persistent mullerian duct syndrome is a rare form of male pseudohermaphroditism. It is usually diagnosed in the early years of life. A case is reported of normal male appearance with bilateral undescended testicles and a right inguinal hernia. On exploration a uterus, two fallopian tubes and a testis were found in the hernial sac. As these patients have an increased risk of gonadal tumours they should be kept under long-term review. PMID- 7492473 TI - Poststroke psychosis. AB - A patient with poststroke psychosis is reported. This is rare but is readily amenable to treatment. Heightened awareness is necessary for early detection and intervention. PMID- 7492474 TI - Collapse of child from undiagnosed diaphragmatic hernia after normal barium investigation. AB - A 16-month-old child who died suddenly from an undiagnosed congenital diaphragmatic hernia is described. She had previously been investigated for persistent vomiting and had undergone a barium study. PMID- 7492475 TI - Postprandial heaviness of the tongue in a diabetic patient revealing myasthenia gravis. PMID- 7492476 TI - Scrotal suppuration after appendicitis. PMID- 7492477 TI - Gastro-esophageal reflux in cerebral palsy. PMID- 7492478 TI - Proceedings of the 4th NAPRTCS Workshop. Los Angeles, 7-9 April, 1994. PMID- 7492480 TI - Chronic dialysis in the infant less than 1 year of age. AB - Dialysis in the infant carries a mortality rate of 16%. Institution of dialysis may be the result of adequate nutritional intake, but avoidance of nutritional intake should never be seen as a way to prevent dialysis. Increased caloric intake, usually via enteral feeding tubes, is needed for optimal growth in the infant with end-stage renal disease (ESRD) in order to attain adequate nutrition with resulting good growth. "Renal" formulae may be constituted as dilute (as in the polyuric infant) or concentrated (as in the anuric infant) to fit the infants needs. Peritoneal dialysis (PD) is the usual mode of renal replacement therapy (97%), with access via a surgically placed cuffed catheter with attention to the placement of the exit site in order to avoid fecal or urinary contamination. PD volumes of 30-40 ml/kg per pass or 800-1,200 ml/m2 per pass usually result in dialysis adequacy. Additional dietary sodium (3-5 mEq/kg per day) and protein (3 4 g/kg per day) are needed, due to sodium and protein losses in the dialysate. Protein losses are associated with significant infectious morbidity and nonresponsiveness to routine immunizations. Hemodialysis (HD) can be performed either as single- or dual-needle access that have minimal dead space (less then 2 ml) and recirculation rate (less then 5%). Attention to extracorporeal blood volume (< 10% of intravascular volume), blood flow rates (3-5 ml/kg per min), heparinization (activated clotting times), ultrafiltration (ultrafiltration monitor), and temperature control is imperative during each treatment.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7492479 TI - Infectious and catheter-related complications in pediatric patients treated with peritoneal dialysis at a single institution. AB - Continuous ambulatory peritoneal dialysis (CAPD) and continuous cycling peritoneal dialysis (CCPD) are the predominant dialytic modalities for the majority of children while awaiting transplantation. Wide acceptability of peritoneal dialysis is hindered by infectious complications. A retrospective review of 367 pediatric patients treated with CAPD/CCPD for at least 3 months from September 1980 through December 1994 revealed that the peritonitis incidence ranged from 1.7 to 0.78 episodes per patient-year. No differences in peritonitis rates were observed between patients treated with CAPD or CCPD. Gram-positive organisms were responsible for the majority of peritonitis episodes. Age, sex, race, primary renal disease, presence of nephrotic syndrome, and serum albumin level were not associated risk factors. Longer time on treatment and diminished serum IgG level were associated with increased peritonitis incidence. Treatment was successfully completed at home in most cases. Almost half of the catheter losses were caused by Staphylococcus, Pseudomonas, and fungal peritonitis and tunnel/exit-site infections. Infectious complications are still the major causes of morbidity and treatment failure in patients treated with CAPD/CCPD. Thus, controlled studies are needed to assess methods for prevention or improvement of peritonitis rates in this patient population. PMID- 7492481 TI - On-line monitoring of the delivery of the hemodialysis prescription. AB - The BioStat 1000 is a new device which employs dialysate-based urea kinetics to calculate the dose of dialysis (Kt/V) based on a two-pool model and protein catabolic rate (PCR). Previous methods relying on blood sampling techniques were subject to error and difficult to implement. This paper describes the features of the Biostat and the results of the first clinical validation study with an early prototype. The BioStat was found to compare favorably with the reference method of direct dialysate quantification (mDDQ) which had been modified to obtain a "two-pool" Kt/V. In 31 patients no significant difference was found between mean Kt/V from the mDDQ and the mean Kt/V from the BioStat (1.35 +/- 0.33 versus 1.38 +/- 0.36, respectively). The PCR was also not significantly different (53.4 +/- 18.5 g/day versus 51.8 +/- 16 g/day, respectively). The BioStat was demonstrated to be a convenient method producing reliable results. PMID- 7492482 TI - Hepatitis C infection in cadaver organ donors: strategies to reduce transmission of infection and prevent organ waste. AB - The transmission of hepatitis C virus (HCV) by organ transplantation has been unequivocally demonstrated. Among recipients of organs from anti-HCV-positive donors, 57%-96% test positive for HCV RNA. Consequently, several organ procurement organizations have adopted a policy restricting the use of anti-HCV positive donors to life-saving transplants (heart, liver, or lung). The differences in the rate of transmission of HCV infection by anti-HCV-positive donors could be related to the prevalence of HCV RNA among these donors. In a national collaborative study of 3,078 cadaver organ donors from eight organ procurement organizations in the United States, the prevalence of anti-HCV antibodies and HCV RNA were 4.2% and 2.4%, respectively. The sensitivity and negative predictive value of anti-HCV antibodies for HCV RNA were 100%. However, despite a specificity of 98.1%, the positive predictive value was only 55.1%. Discarding organs from enzyme-linked immunosorbent assay 2-positive donors would eliminate transmission, but organs from 1.88% of donors would be wasted. Clinical and laboratory characteristics did not distinguish anti-HCV-positive donors with and without HCV RNA. Hence, to reduce waste it is necessary to develop confirmatory tests with a higher specificity for HCV RNA than those that are currently available. Even if anti-HCV-positive but HCV RNA-negative donors could be identified and utilized, 2.4% of cadaver organ donors that test positive for serum HCV RNA would remain unsuitable for transplantation of non-life-saving organs. Hence, several authors have suggested the use of kidneys from anti-HCV positive donors in recipients with pre-existing HCV infection.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7492483 TI - Transmission of hepatitis C virus by kidney transplantation: impact of perfusion techniques and course of viremia post transplant. AB - Hepatitis C virus (HCV) infection is the leading cause of post-transplant non-A, non-B hepatitis. Although many end-stage renal disease patients present for transplantation already infected with HCV, some recipients acquire the infection by transmission from the donor organ. We have detected serological evidence for HCV infection in 6.8% of our organ donors using second-generation anti-HCV assays. Approximately one-third of the patients who received an organ from a HCV carrier donor developed chronic transaminasemia and 8 of 14 (56%) patients converted from HCV RNA negative to positive in the posttransplant period. Demonstration of the course of viremia and transaminasemia is presented for 2 patients in whom transmission of HCV occurred. Using pulsatile machine perfusion, we were able to demonstrate that a standard perfusion of 20 h reduced the viral load in the kidney by 75%, additional flushes and a subsequent perfusion reduced the total viral titer by more than 99%. Thus, although transmission of HCV does occur with solid-organ transplantation, differences in the incidence of transmission between centers may be related to techniques of organ preservation. PMID- 7492484 TI - Correlation of donor antigen-specific hyporeactivity with allogeneic microchimerism in kidney and lung recipients. AB - Our previous studies indicate donor antigen-specific hyporeactivity is a useful marker for identifying solid organ transplant recipients at low risk for immunological complications; the hyporeactive subgroup experiences a lower incidence of chronic rejection. One purpose of the current study was to determine whether hyporeactivity could be identified in pediatric kidney recipients and whether it correlated with improved graft outcome. Of 18 pediatric kidney recipients tested, 6 (33%) had developed donor antigen-specific hyporeactivity. All 18 experienced good graft outcome. Second, we determined whether donor antigen-specific hyporeactivity correlates with peripheral blood microchimerism and outcome in adult kidney recipients. Our previous studies of lung recipients demonstrated development of obliterative bronchiolitis in recipients with microchimerism who remain responsive, but not in recipients who had become hyporesponsive to donor antigen. Preliminary results in our current study of 23 adult kidney recipients indicate microchimerism for 6 (26%): 4 hyporesponsive and 2 responsive to donor antigen. Microchimerism was not detected for 17 recipients: 6 hyporesponsive and 11 responsive to donor antigen. One hyporesponsive/chimeric patient and 4 recipients negative for both parameters have been diagnosed with biopsy-proven chronic rejection. In summary, both hyporeactivity and chimerism are found at a higher frequency in lung than kidney recipients. Unlike lung recipients, not all hyporesponsive kidney recipients had peripheral blood chimerism. Additional numbers are needed to determine if microchimerism correlates with donor antigen-specific hyporeactivity or graft outcome. PMID- 7492485 TI - Living-related donor transplants should be performed with caution in patients with focal segmental glomerulosclerosis. AB - The success rates of living-related donor (LRD) transplants are clearly superior to those obtained with cadaver donors. However, caution should be exercised when considering LRD transplantation for a condition which has an increased chance of recurring after transplantation and causing ultimate graft failure. The recurrence rate of focal segmental glomerulosclerosis (FSGS) in the allograft is 20%-40%, with graft failure resulting in 40%-50% of these cases. However, these figures may be an underestimation of the true rate of recurrence of FSGS. Once a first transplant fails due to recurrent disease, the risk of recurrence in the second transplant approaches 80%. Subgroups of patients at high risk for recurrence have been identified. In patients not at high risk for recurrent FSGS, the use of a LRD should be considered, provided that the donor and recipient and their families have been informed that the disease may recur and lead to graft failure. In patients at high risk for recurrence, a LRD transplant should be avoided. Hopefully, future development of a simple and reliable test to predict the likelihood of recurrence will enable us to counsel and advise our patients with FSGS about the wisdom or dangers of proceeding with a LRD transplant. PMID- 7492486 TI - FK506 in pediatric kidney transplantation--primary and rescue experience. AB - Between 14 December 1989 and 17 December 1993, 43 patients undergoing kidney transplantation alone at the Children's Hospital of Pittsburgh received FK506 as the primary immunosuppressive agent. The mean recipient age was 10.2 +/- 4.8 years (range 0.7-17.4 years), with 7 (16%) children under 5 years of age and 2 (5%) under 2 years of age. Fifteen (35%) children underwent retransplantation, and 5 (12%) had a panel-reactive antibody level greater than 40%. Twenty-two (51%) transplants were with cadaveric donors and 21 (49%) were with living donors. The mean follow-up was 25 +/- 14 months; there were no deaths; 1- and 3 year actuarial graft survival was 98% and 85%. The mean serum creatinine and blood urea nitrogen were 1.2 +/- 0.6 mg/dl and 26 +/- 11 mg/dl; the calculated creatinine clearance was 75 +/- 23 ml/min per 1.73 m2. Twenty-four (62%) patients have been successfully withdrawn from steroids and 24 (62%) require no anti hypertensive medication. Improved growth was seen, particularly in pre-adolescent children off steroids. Between 28 July 1990 and 2 December 1993, 24 children were referred for rescue therapy with FK506, 14.6 +/- 16.4 months (range 1.1-53.2 months) after transplantation. Nineteen (79%) were referred because of resistant rejection; 4 (17%) were referred because of proteinuria; 1 (4%) was switched because of steroid-related obesity. There were no deaths; 1- and 2-year graft survival was 75% and 68%; 17 (71%) patients were successfully rescued, including 1 of 2 patients who arrived on dialysis; 4 (24%) of the successfully rescued patients were weaned off steroids.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7492487 TI - A clinical trial of cyclosporine G in cadaveric renal transplantation. AB - Cyclosporine G (OG 37-324) reportedly is an efficacious immunosuppressant with less nephrotoxicity than cyclosporine A. This is a prospective randomized double blinded trial comparing cyclosporine G and cyclosporine A in cadaveric renal transplantation. Patient and graft survival, as well as major infectious complications, were not different between the two groups. Objective parameters of renal function, including serum creatinine, creatinine clearance, and inulin clearance, were routinely performed. These generally demonstrated less nephrotoxicity in those patients treated with cyclosporine G compared with cyclosporine A. Minor elevations of alanine aminotransferase were noted in the cyclosporine G-treated patients but this was not associated with acute morbidity. Overall, cyclosporine G appears to be equally as effective as cyclosporine A, but demonstrated notably less nephrotoxicity. PMID- 7492488 TI - Brequinar sodium. AB - Brequinar sodium is one of a new group of immunosuppressive drugs currently being developed for the prevention of organ graft rejection. The preclinical and clinical characterization of brequinar sodium has clearly demonstrated that this drug is a compound with highly effective immunosuppressive activity that is the result of its interference with pyrimidine biosynthesis and the disruption in cell proliferation. The antiproliferative activity of the drug influences both T and B cell immune function, thereby providing the potential for the use of this compound in combination with other immunosuppressive agents for more specific and effective prevention of allograft and xenograft rejection. Experimental studies with brequinar and cyclosporin or FK 506 have shown that the use of these agents in combination provides a synergistic effect for the prevention of graft rejection, and studies are now in progress to evaluate the application of this drug in clinical transplantation. PMID- 7492489 TI - Renal transplant fine needle aspiration and cytokine gene expression. AB - Fine needle aspiration of renal transplants is a safe effective means of obtaining intragraft material for diagnostic evaluation of transplant dysfunction in the first 3-6 months post engraftment. Aspiration can be performed daily as an outpatient to assess causes of renal insufficiency and response to therapeutic intervention. Aspiration also provides a unique tool for the investigation of pathogenetic mechanisms in acute rejection; samples can be subjected to reverse transcriptase-polymerase chain reaction with quantitation of cytokine gene expression. We have utilized this technique to demonstrate the strong, likely causal, relationship between gamma-interferon and acute rejection. The ability to perform aspirates frequently affords an opportunity to map the cytokine cascade prior to and during episodes of acute rejection. This may identify new areas for therapeutic intervention during the process of rejection. PMID- 7492491 TI - Do children exhibit catch-up growth post transplant: North American Pediatric Renal Transplant Cooperative Study special study. AB - Changes in height deficit and standard deviation score (SDS) were evaluated in 524 recipients who were growth retarded at transplantation (SDS > -2.00) and were followed for at least 2 years post transplant. At 2 years the delta SDS was 0.32 +/- 0.04 and the delta height deficit was 0.75 +/- 0.23. Therefore despite improvement in the SDS at 2 years post transplant, the change in height deficit was < 1 cm. Change in height deficit may be a better indication of "catch-up" growth following transplantation. PMID- 7492490 TI - An analytical review of growth hormone studies in children after renal transplantation. AB - With steady improvement in 1- and 5-year patient and graft survivals in the last decade, rehabilitation of the child is the major focus of the transplant physician. The notion that the elimination of the uremic milieu should enable children to grow has not been born out over time, and growth retardation continues to be a serious morbidity in many children despite a well-functioning renal allograft. In children with chronic renal failure prior to renal transplantation, recombinant human growth hormone (rhGH) has been recently shown in controlled trials to improve growth. The use of rhGH in children after renal transplantation is controversial, since uncontrolled studies have questioned its safety. Acute rejection and graft loss have been reported in children after the initiation of rhGH. This study analyzes the data regarding the safety and efficacy of rhGH in children after renal transplantation as presented in seven current published reports. The demographic characteristics of the patients, the dose and duration of rhGH therapy, the "growth" rates of the patients before and after rhGH therapy, renal function before and after rhGH therapy, and other possible complications are reviewed. Based on this analysis, suggestions for future studies are made. PMID- 7492492 TI - On-line dialyser clearance using conductivity. AB - A method allowing non-invasive measurement of dialyser clearance in vivo is described. Electrolyte concentration difference is measured by two conductivity sensors positioned up- and downstream of the dialyser in the dialysate path. From the conductivity differences measured at two different dialysate input concentrations and dialysate flow, electrolyte dialysance is calculated. The method takes recirculation effects into account and measures effective clearance rather than dialyser clearance. Test results comparing on-line electrolyte clearance with blood side clearance corrected for recirculation are consistent with this calculation. From clearance measured several times during dialysis and time on dialysis, K.t can be calculated with an estimated accuracy of +/- 10%. This method can be used to control the efficacy of each individual treatment. The K.t goal can be derived from monthly two-pool kinetic modelling. PMID- 7492493 TI - New way of working has 'vastly improved' the quality of care delivered to patients. PMID- 7492494 TI - Fighting to win equality. PMID- 7492496 TI - A shared strategy. PMID- 7492495 TI - Increasing independence. PMID- 7492497 TI - Occupational asthma: assessing the risk. AB - Asthma caused by factors encountered at work can be overlooked. The author describes some of the risk factors and occupational groups involved and how nurses can help to detect cases of asthma caused by materials found at work. PMID- 7492498 TI - Bionursing: assessment of stress incontinence. AB - The fifth article in our series on bionursing uses a case study based on fact, with personal details altered to maintain confidentiality, to illustrate the importance of biological theories underpinning nursing care. The authors describe how perceptions of the ageing process can cloud nurses' judgement. They emphasise the need for staff to include physiological factors in their assessment of patients. PMID- 7492499 TI - Health secretary Stephen Dorrell has pledged to reduce NHS bureaucracy. PMID- 7492500 TI - Perceptions of patients' pain: a study assessing nurses' attitudes. AB - This study of 35 orthopaedic nurses assessed attitudes to pain and its relief. Using an anonymous questionnaire, nurses gave their views on a range of issues from what patients' expectations of post-operative pain should be, to the use and effectiveness of pain assessment tools. The findings suggest that nurses require re-education in various aspects of pain and analgesia provision to ensure that patients do not feel pain unnecessarily and receive appropriate pain relief promptly. The study recommends that pain assessment tools are used by orthopaedic nurses and that further training is required in the pharmacology of analgesic agents. PMID- 7492501 TI - Allowed to die. PMID- 7492502 TI - Love your managers. PMID- 7492503 TI - A shared formulary is good for us all. PMID- 7492504 TI - Change and a growing crisis. PMID- 7492505 TI - Nursing must look after its young. PMID- 7492506 TI - Second class care. PMID- 7492507 TI - How to do well at exams. PMID- 7492508 TI - No rules in the Home Office lottery stakes. PMID- 7492509 TI - Overseas students sent home after training. PMID- 7492512 TI - It's good to talk. PMID- 7492511 TI - 'Stay nursing, stay motivated'. PMID- 7492510 TI - Elsie's story. PMID- 7492513 TI - Euthanasia and the nurse. PMID- 7492514 TI - Cervical screening: a positive view. PMID- 7492515 TI - Exposure of macrophages to PLA2-modified lipoproteins leads to cellular lipid accumulations. AB - After a 24-h exposure of mouse peritoneal macrophages to LDL, of which the phospholipids were reduced to 30% by phospholipase A2 (PLA2)-treatment, the cellular level of free and esterified cholesterol was elevated 1.9- and 5.0-fold, respectively. Furthermore, the activity of the cytosolic acyl-CoA cholesterol acyl-transferase (ACAT), which was calculated from the rate of 14C-oleate incorporation into the cellular cholesteryl esters, increased with the degree of LDL hydrolysis. The incubation of macrophages with native HDL led to decreased labeling of cellular cholesteryl esters. However, after PLA2-treatment of HDL the esterification rate increased with the degree of hydrolysis analogous to PLA2 modified LDL. The formation of numerous intracellular lipid droplets was observed by light microscopy after staining with Sudan-III. These data suggest that phospholipases A2 may play a role in the transformation of macrophages into foam cells, a hallmark of early atherosclerotic lesions. PMID- 7492516 TI - Interest of argyrophilic proteins nucleolar organizer regions (AgNOR) to estimate the reactivity of T cell clones against autologous malignant B-NHL cells. AB - The silver-stained acidic proteins of interphase nucleolar organizer regions (AgNOR) were studied to assess the reactivity of 11 T cell clones (Tcc) against autologous B-NHL cells. Tcc, derived from tumour-infiltrating T lymphocytes of seven patients, were cultured in the presence of irradiated autologous B-NHL cells with recombinant IL-2. Then the percentage of activated T cells expressing the CD25 antigen and their proliferating rate (measured by [3H]thymidine incorporation) were estimated. Simultaneously, at the end of this culture period, B-NHL cells were eliminated by using a cell-sorter, and the resulting purified T cells were studied for AgNOR expression. Tcc cultured without B-NHL cells served as controls. In eight out of the 11 Tcc, the increase in [3H]thymidine incorporation and CD25 expression paralleled the increase of AgNOR area and number. By contrast, in the three remaining Tcc, we observed a decrease (one Tcc) or an increase (two Tcc) of AgNOR parameters, whereas CD25 expression and/or [3H]thymidine incorporation remained unchanged in comparison to control cultures. We concluded that quantification of AgNOR should be a more sensitive technique than thymidine incorporation and CD25 expression for detecting the activation in vitro of T cells induced by autologous B-NHL cells. PMID- 7492517 TI - Immunohistochemical detection of alpha 1-antitrypsin, alpha 1-antichymotrypsin, transferrin and ferritin in ameloblastoma. AB - An immunohistochemical study was performed to investigate the presence of alpha-1 antitrypsin (alpha 1-AT), alpha-1-antichymotrypsin (alpha 1-ACT), transferrin and ferritin in 32 ameloblastomas and to evaluate the co-expression of these antibodies. Histologically, we recognized the following 6 patterns in the series of 32 ameloblastomas and at least 2 patterns in variable proportions were present in each of our cases: follicular pattern (21 cases, 66%), plexiform pattern (17 cases, 53%), cystic pattern (21 cases, 66%), acanthomatous pattern (10 cases, 31%), granular cell type (2 cases, 6%), and hyalinized stromal pattern (20 cases, 63%). Neoplastic epithelia of cystic pattern were divided into superficial cell, basal cell and whole layer to compare the immunohistochemical localization. The results made on the various patterns of ameloblastomas were as follows: (1) alpha 1-AT positivity in plexiform, cystic and hyalinized stromal patterns was significantly higher than that of alpha 1-ACT (P < 0.05). (2) The incidence of transferrin in follicular and plexiform patterns was markedly higher than that of ferritin in the same patterns (P < 0.025 and P < 0.01). Transferrin strongly stained metaplastic squamous cells of acanthomatous pattern and basal cells of cystic epithelium. (3) Granular cells reacted with transferrin and ferritin. (4) In follicular and acanthomatous patterns, coexpression of alpha 1-AT and alpha 1 ACT, alpha 1-AT and transferrin, or alpha 1-ACT and transferrin was higher than that of another combination. On the other hand, co-expression of alpha 1-AT and transferrin in plexiform and cystic patterns was higher than that of other antibodies. These results of co-expression of 4 antibodies used in the present study, suggest that the histogenesis of follicular and acanthomatous patterns is different from that of plexiform and cystic patterns. PMID- 7492519 TI - Combined malignancy associated change and contextual analysis for computerized classification of cervical cell monolayers. AB - Previous efforts to automate cervical cell analysis have focused on algorithms that use measurements of high resolution images of individual cells for 'rare event' detection of abnormal cells, sometimes in combination with cluster analysis of low resolution images. A new methodology was developed in which intermediate cell markers (indicating malignancy associated change) and contextual analysis of slide architecture were performed on the same digital images at 0.33 mu pixel resolution. A total of one hundred and forty-six cases that were either negative or at least high grade squamous intraepithelial lesions (HGSIL) were prepared as monolayers, and about 20 fields per slide were analyzed. Features most important for intermediate cell marker analysis were those that measured the variation in nuclear texture measurements across the slide as well as densitometric features. The most discriminatory contextual features were those that measured the variation of the arrangement of cells within a cluster across the slide. Linear discriminants were calculated using the most important features from both types of analysis; they provided smear classification accuracies of 71 86%. PMID- 7492520 TI - Aneuploidy occurrence in human tumours: a logical-automaton approach. AB - The search for new, reliable factors of prognosis in cancerology is sadly deficient at the present moment. Of these factors, the measurement of ploidy gives rise to considerable hope. Nevertheless, despite the impressive number of papers currently published, no general law seems to be emerging that associates the ploidy rate of a tumour with its clinical evolution in a patient. The purpose of the present work is firstly to use a logical automation to describe the cell cycle in terms of binary variables (the validation of the methodology), and secondly to demonstrate that a certain 'cancer logic' can be distilled, at least with respect to the genesis of DNA histograms among tumours. PMID- 7492521 TI - Segmenting textures using cells with adaptive receptive fields. AB - Textural segmentation plays an important role in the figure-ground discrimination process. Evidence from neuroscience and psychophysics suggests that the segregation of texture patterns composed of oriented line segments is strongly influenced by the orientation contrast between the patterns (Nothdurft, Vision Res. 31, 1073-1078, 1991). In contrast to models available in the literature, this paper presents a neural network architecture for textural segmentation that can adaptively delimit the boundaries of uniformly textured regions. Cells with adaptive receptive fields encode uniformly textured regions by diffusively interpolating the estimates of feature orientations across the image. Orientation contrast boundaries are detected at gradients across the interpolated regions to produce the final segmentation. Consistent with neurophysiological data from simple and complex cells sensitive to static and moving textural patterns (Hammond and MacKay, Exp. Brain Res. 30, 275-296, 1977), the present model suggests that preattentive textural segregation can be performed by early visual processes localized to areas V1, V2, and perhaps V3 or V5. The computational results further support the idea that textural segmentation should not be thought of as a 'static' process, but as a system that employs cells with context dependent response characteristics (Gilbert and Wiesel, Vision Res. 30, 1689 1701, 1990), here modeled as adaptive receptive fields. PMID- 7492518 TI - Immunohistochemical correlation of steroid receptors and disease-free interval in 206 consecutive cases of breast cancer: validation of telequantification based on global scene segmentation. AB - Objective algorithms to evaluate immunocytochemistry in cytology and in histological sections are still under investigation. Clinical value, simplicity, robustness and adaptation to new technologies such as telemedicine and telequantification have to be taken into consideration. The present paper summarizes the experience of the immunoquantification of 206 cases diagnosed from 1990 to 1991, 36 clinical stage I cases, 110 stage II cases and 60 stage III cases. Initial clinical stage IV cases were excluded from the series. Quantification was done on frozen sections incubated with monoclonal estrogen and progesterone receptors (ER, PgR) by the peroxidase antiperoxidase (PAP) technique revealed with DAB. Automatic computerized measurements with a 550-nm filter were based on global scene segmentation, obtaining the parameters of percentage of positive area (pA) and immunoscore (ISC = sum i x pi), being the staining intensity (i) automatically evaluated in 5 grades multiple of the standard deviation (SD) of the control specimen (i x SD) stained with progressive haemathoxyline. Cut point of positive and negative specimens regarding ISC were obtained by semilogarithmic plot with biochemical receptor values (r = 0.73). The whole series showed a significant increase of negative specimens according to clinical stage and a higher number of PgR negative cases (44%) than ER negative cases (32%). The clinical relevance of the quantified parameters were evaluated by free interval disease survival curves being significant in the positive cutting point of ISC = 100 (ER P < 0.06; PgR P < 0.02) and the 40% negative area for the pA (ER P < 0.001; PgR P < 0.004). This quantification technique, which does not require individual cell segmentation, was reproducible at a distance (telequantification through high resolution videophones at 9600 bauds transmission rate; 55 cases). Coefficient of variation (CV) of the parameters in the original and transmitted cases were in the range of 0-5% in specimens with positive areas over 70%; less positive specimens require lower segmentation thresholds to get acceptable coefficients of variation. According to our results, immunohistochemical steroid receptor contents in breast cancer pathology, quantified by the technique of pixel integration, demonstrated the added value of the percentage of positive area in patient follow-up. Those having more than 40% of negative areas have recurrences mainly within the first year. PMID- 7492522 TI - Separating colour and luminance information in the visual system. AB - In our visual world we can distinguish with ease between chromatic and luminance contrasts. However, in our retinae most neurones are responsive to both chromatic and luminance changes and therefore send ambiguous or 'multiplexed' information to the higher visual centres. Psychophysical evidence suggests that some cortical process must subsequently separate out this information into its chromatic and luminance components. The purpose of this communication is to review and critically evaluate the different existing schemes for doing this. To assist in this evaluation a linear systems analysis is employed in which model cortical neurones are imputed with the property of providing information about either colour or luminance. It is concluded that there is currently no unified scheme available to explain a separation of colour and luminance information in the visual system. Some theoretical considerations and most promising approaches to solving the problem are noted, but it is suggested that there may be definite limits to the ability of the visual system to achieve complete separation of colour and luminance from the retinal signal. PMID- 7492524 TI - Deriving channel gains from large-area sine-wave contrast sensitivity data. AB - A wealth of detection data can be accounted for by a spatial-vision model including a finite number of space-variant, spatial-frequency and orientation selective channels of varying gains coupled with a detection rule involving probability summation over space and among channels. This paper shows that the detection of large-area, foveally fixated sine-wave gratings can be understood as if it occurred merely as a result of the activity of the subset of channels whose orientation matches that of the gratings, and operating under a peak-detection rule. This simplification makes it possible to show the theoretical relationship between the large-area sine-wave contrast sensitivity function, the channel gain function, and the channel modulation-transfer functions. It is also shown that the human visual system must have many more channels than are normally assumed in spatial-vision models, for otherwise the contrast sensitivity function would show significant bumps. An unlimited-channel model with a given mathematical form for the channels' modulation transfer functions is used to derive a mathematical expression for the (foveal) channel gain function from the large-area psychophysical sine-wave contrast sensitivity function. PMID- 7492523 TI - Grating detection and identification dissociated by pattern adaptation. AB - It two gratings differ in a higher harmonic only, grating identification is expected to depend on that harmonic detection. Yet the comparison of data from several studies suggests that pattern adaptation might differently affect detection and identification. In order to verify this dissociation, a study was made of the effects of adaptation to complex gratings and their harmonics on the detection of the harmonics and on the identification of the complex gratings. Square-wave, first-plus-third-harmonic and missing-fundamental complex gratings were used. The threshold was measured either by the two-interval forced-choice method or by the one-interval method of constant stimuli. For all types of stimuli and procedures, the detection threshold was mostly elevated after adaptation to the corresponding harmonic and the identification threshold was mostly elevated after adaptation to the complex stimulus. This dissociation could only partly be related to the presence of higher harmonics or to the psychophysical procedure. Analysis of threshold changes by adaptation and of phenomenal stimulus appearance suggests that both local features and more global features might contribute to grating identification. PMID- 7492525 TI - Motion detectors and motion segregation. AB - The response of motion detectors necessarily confound image velocity with image structure. In particular, even a rigidly moving image (with a uniform velocity field) will give rise to non-uniform detector responses. A mathematical framework has been proposed on how to intrinsically compare motion detectors' responses so that their differences will reflect the true differences in image velocity (Zhang and Wu, Proc. Natl. Acad. Sci. USA 87, 7819-7823, 1990). Here, this notion of 'intrinsic differentiation' was implemented by introducing a gamma-matrix determined by the image spatial gradients. The perceptual phenomenon of random dot motion segregation was successfully simulated. PMID- 7492526 TI - Eye movements and their role in visual and cognitive processes. PMID- 7492527 TI - The role of visual and cognitive processes in the control of eye movement. PMID- 7492528 TI - The role of eye movement in the detection of contrast and spatial detail. PMID- 7492529 TI - Binocular eye movements and the perception of depth. PMID- 7492531 TI - The role of eye movements in the perception of motion and shape. PMID- 7492530 TI - Eye movement and visual localization of objects in space. PMID- 7492532 TI - Comparison of perception in the moving and stationary eye. PMID- 7492533 TI - Eye movements in visual search: cognitive, perceptual and motor control aspects. PMID- 7492534 TI - Eye movements and reading. PMID- 7492535 TI - Eye-movement models for arithmetic and reading performance. AB - Three stochastic eye-movement models for arithmetic and reading performance have been proposed, one for arithmetic and two for reading. Each model characterizes a real-time stochastic process in terms of fixation durations and saccadic movement, but only direction and length of saccades are considered, not acceleration or velocity. Aspects of the models that are emphasized, partly because of their general neglect in the literature, are the probability distribution of fixation durations and the random walk of saccade directions. The distributions of fixation duration are approximately exponential, but systematic deviations can be accounted for in the models, even though the fit to data is not perfect. In the case of the arithmetic algorithms of addition and subtraction, the random walk of the normative model has only two possible moves. Data are also presented on backtracking, skipping and wandering eye movements, each of which has a significant relative frequency. The first reading model is called a minimal control model, because it does not take account of the effects of many local variables, e.g., word length, that have been extensively studied. The axioms on fixation duration for the minimal control model are the same as for the arithmetic model. Abstracting from the different arrangement of stimuli in arithmetic algorithms and in linear text, the axioms on saccadic motion for the two models are also essentially identical. The stochastic nature of both models is strongly supported by data on the independence of fixation durations from previous fixation durations. Additional detailed evidence is presented for the arithmetic model. To better account for a great variety of experimental results concerning significant effects on eye movements in reading, a text-dependent probabilistic model of reading is introduced. Significant local effects fall into three classes, identified as line, word and grammatical variables. The revised axioms embody five features of text known to be significant: (i) fixation duration depends on the number of letters in a word; (ii) a saccade is longer when a longer word is to the right; (iii) a saccade is longer when the current fixation is on a longer word; (iv) high-frequency fixation words have the highest probability of being skipped; (v) ambiguous or difficult grammatical structures increase backtracking. PMID- 7492536 TI - Predictive control of eye movement. PMID- 7492537 TI - Effects of geometric isomerism and ligand substitution in bifunctional dinuclear platinum complexes on binding properties and conformational changes in DNA. AB - The DNA binding profile of a series of dinuclear platinum complexes [{trans-PtCl (L)2}2H2N(CH2)nNH2]2+ (L = NH3 or py; 1,1/t,t/NH3 and 1,1/t,t/py, respectively) and [{cis-PtCl-(NH3)2H2N(CH2)nNH2]2+ (1,1/c,c/NH3) was examined to compare the effects of geometrical isomerism and the presence of ligands other than NH3 in the coordination sphere. Steric effects, because of the geometry of the leaving groups cis to the diamine bridge or the presence of planar pyridine ligands, result in diminished binding to calf thymus DNA for these isomers. In contrast, the pyridine derivative shows a distinct binding preference for poly(dG dC).poly(dG-dC) in comparison to both NH3 isomers. Both NH3 complexes induced the B-->Z transition in poly(dG-dC).poly(dG-dC), but the presence of a pyridine ligand stabilized the B conformation. The bifunctional binding of the NH3 isomers results in unwinding of supercoiled pUC19 plasmid DNA equivalent to cis-DDP, while the unwinding of the pyridine derivative is approximately twice that of the mononuclear trans-[PtCl2(py)2]. DNA-DNA interstrand cross-linking is very efficient for all three agents, but sequencing studies indicated that only the 1,1/t,t/NH3 derivative is capable of forming a (Pt,Pt) intrastrand cross-link to the adjacent guanines of a d(GpG) sequence. The effects on DNA caused by bifunctional binding of dinuclear complexes are compared with those from the mononuclear [PtCl2(NH3)2] isomers. The results are discussed with respect to the antitumor activity of the dinuclear series. PMID- 7492540 TI - The conserved seven-transmembrane sequence NP(X)2,3Y of the G-protein-coupled receptor superfamily regulates multiple properties of the beta 2-adrenergic receptor. AB - The beta 2-adrenergic receptor (beta 2AR) is a member of a large superfamily of seven transmembrane domain, G-protein-coupled receptors. Within the putative seventh transmembrane domain of the beta 2AR is a sequence of amino acids, NPLIY, which is conserved with minor variations in all members of the superfamily. Previously it was demonstrated that mutation of tyrosine residue 326 to an alanine abolished agonist promoted sequestration of this mutant without affecting its ability to maximally stimulate adenylyl cyclase in membranes [Barak, L.S., Tiberi, M., Freedman, N.J., Kwatra, M.M., Lefkowitz, R.J., & Caron M.J. (1994) J Biol. Chem. 269, 2790-2795]. In the present study we characterized the NPLIY amino acid sequence in an attempt to determine how it can affect the agonist mediated sequestration of the beta 2AR and to test whether it is a functional motif. We find that point mutations of the most conserved amino acids, N, P, and Y, in this sequence affect several other receptor properties in addition to sequestration. Mutation of asparagine 322 to an alanine resulted in complete uncoupling of the receptor, loss of high-affinity agonist binding, and abolition of receptor sequestration, down-regulation, and phosphorylation. In contrast, a conservative mutation of this residue to an aspartic acid (as found in the thrombin receptor) resulted in an improvement of G-protein coupling without adversely affecting other receptor properties. Substitution of proline residue 323 with an alanine residue resulted in a receptor with mild deficits in sequestration and coupling, a reduced agonist-mediated phosphorylation, and no change in down-regulation.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7492539 TI - Net sugar transport is a multistep process. Evidence for cytosolic sugar binding sites in erythrocytes. AB - Human erythrocyte net sugar transport is hypothesized to be rate-limited by reduced cytosolic diffusion of sugars and/or by reversible sugar association with intracellular macromolecules [Naftalin, R.J., Smith, P.M., & Roselaar, S.E. (1985) Biochim. Biophys. Acta 820, 235-249]. The present study examines these hypotheses. Protein-mediated 3-O-methylglucose uptake at 4 degrees C by human erythrocytes and by resealed, hypotonically lysed erythrocytes (ghosts) is inhibited by increasing solvent viscosity. Protein-mediated transport and transbilayer diffusion of the slowly transported substrate 6-NBD glucosamine are unaffected by increasing solvent viscosity. These findings suggest that protein mediated 3-O-methylglucose transport is diffusion-limited in erythrocytes. More detailed analyses of red cell 3-O-methylglucose uptake (at 4 degrees C and at limiting extracellular sugar levels) reveal that net influx is a biexponential process characterized by rapid filling of a small compartment (C1 = 29 +/- 6% total cell volume; k1 = 7.4 +/- 1.7 min-1) and slow filling of a larger compartment (C2 = 71 +/- 6% total cell volume k2 = 0.56 +/- 0.11 min-1). Erythrocyte D-glucose net uptake at 4 degrees C is also a biphasic process. Transmembrane sugar leakage is a monoexponential process indicating that multicomponent, protein-mediated uptake does not result from sugar uptake by two cell populations of differing cellular volume. Sugar exit at limiting 3-O methylglucose concentrations is described by single exponential kinetics. This demonstrates that multicomponent sugar uptake does not result from influx into two populations of cells with widely different sugar transporter content. We conclude that biexponential sugar uptake results from slow (relative to transport) exchange of sugars between serial, intracellular sugar compartments. Biexponential sugar uptake is observed under equilibrium exchange conditions (intracellular sugar concentration = extracellular sugar concentration) but only at 3-O-methylglucose concentrations of less than 1 mM. Above this sugar concentration, exchange uptake is a monoexponential process. Because diffusion rates are independent of diffusant concentration, this suggests that multicomponent uptake results from high-affinity sugar binding within the cell. The concentration of cytosolic binding sites (30 microM, Kd(app) = 400 microM) was estimated from the equilibrium cellular 3-O-methylglucose space. Biexponential net 3-O-methylglucose uptake is also observed in human erythrocyte ghosts, in control human K562 cells, and in K562 cells induced to synthesize hemoglobin by prolonged exposure to hemin. This demonstrates that neither membrane-bound nor free cytosolic hemoglobin forms the sugar-binding complex.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 7492538 TI - Conformational changes and calcium binding by calretinin and its recombinant fragments containing different sets of EF hand motifs. AB - Four recombinant fragments, representing different sets of EF-hand motifs of rat calretinin (CR) (I-II, I-III, III-VI, IV-VI), were prepared, and their Ca2+ induced conformational changes were compared with those of full-length recombinant CR. All fragments were able to bind calcium ions as shown by 45Ca2+ overlay method on nitrocellulose and fluorescence measurements. The intrinsic tryptophan fluorescence intensity (FI) of apo-CR reversibly increased about 3 fold upon addition of calcium, indicating a change of conformation. The FI of fragments I-II (Trp 25) and I-III (Trp 25 and 116) increased about 1.4-fold on calcium binding, but that of fragment III-VI (Trp 116) increased 3.5-fold. Calcium titration of CR monitored by Trp fluorescence intensity showed that recombinant CR and some fragments bound Ca2+ with high affinity (Kd below 0.4 microM) and with high cooperativity. An apparent Hill coefficient for Ca2+ induced fluorescence changes in CR was about 3.7. CR bound to organomercurial agarose Cys 101 and 266 did not form cystine. The fluorescence intensities of cysteine-linked fluorescent probes 5-iodoacetamidofluorescein and N-(1 pyreneiodoacetamide) were increased approximately 1.3-fold upon calcium binding by CR. These data indicate that CR binds Ca2+ with high affinity and cooperativity and that this binding induces a change of conformation that involves the interaction of different parts of the molecule. Taken together, our results suggest that CR works as an on/off switch within a narrow range of free Ca2+ by interacting with as yet unidentified targets. PMID- 7492541 TI - Prodomain mutations at the subtilisin interface: correlation of binding energy and the rate of catalyzed folding. AB - The in vivo folding of subtilisin is dependent on a 77 amino acid prosequence, which is eventually cleaved from the N-terminus of subtilisin to create the 275 amino acid mature form of the enzyme. The recent determination of the structure of a complex of the prodomain and a calcium-free subtilisin mutant has suggested how the prodomain may catalyze subtilisin folding [Bryan, P., Wang, L., Hoskins, J., Ruvinov, S., Strausberg, S., Alexander, P., Almog, O., Gilliland, G., & Gallagher, T. (1995) Biochemistry 34, 10310-10318]. In the complex, the prodomain packs against the two parallel surface helices of subtilisin (residues 104-116 and residues 133-144) and supplies caps to the N-termini of the two helices. The binding site is contained almost entirely in the linear sequence 100-144 of subtilisin. The C-terminus of the prodomain (residues 72-77) extends out from its central part to bind like a substrate in subtilisin's active site cleft. The simplest model of catalyzed folding is one in which the observed binding interaction in the complex accelerates folding by stabilizing an intermediate which includes the 45 amino acid alpha beta alpha substructure in subtilisin. According to our hypothesis, amino acids 100-144 would have a native-like fold in the intermediate which the prodomain stabilizes. Guided by the structure of the bimolecular complex of subtilisin and its prodomain, we have constructed mutations in the C-terminal region of the prodomain. Analysis of five mutants reveals a general correlation between the ability of the prodomain to bind to native subtilisin and its ability to accelerate subtilisin folding.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7492542 TI - Binding to protein targets of peptidic leads discovered by phage display: crystal structures of streptavidin-bound linear and cyclic peptide ligands containing the HPQ sequence. AB - The streptavidin-bound crystal structures of two disulfide-bridge cyclic peptides (cyclo-Ac-[CHPQGPPC]-NH2 and cyclo-Ac-[CHPQFC]-NH2) and of a linear peptide (FSHPQNT) were determined, as well as the structure of apostreptavidin (streptavidin-sulfate). Both the linear and disulfide-bridged cyclic peptides studied share a common HPQ conformation and make common interactions with streptavidin, although significant differences in structures and interactions occur for flanking residues among the complexes. The conformation of the linear peptide in the crystal structure of streptavidin-FSHPQNT was found to differ from that in the same complex published [Weber, P. C., Pantoliano, M. W., & Thompson, L. D. (1992) Biochemistry 31, 9350-9354]. In the present investigation, the HPQNT portion of the ligand is well-defined with some density defining the Phe, whereas in the investigation of Weber et al. only the HPQ segment of the bound peptide could be interpreted. Both bound cyclic peptides adopt a beta-turn involving an H bond between the His main chain carbonyl and the main chain amide NH of the i+3 residue. In the streptavidin-bound cyclo-Ac-[CHPQFC]-NH2 structure, there is an additional H-bond, indicative of alpha-helix, between the main chain His carbonyl and the main chain C-terminal Cys amide NH group. Binding interactions for both cyclic and linear peptides include direct H-bonds, H-bonds mediated by tightly bound water molecules, and hydrophobic interactions. The above structures and that of streptavidin-biotin in the literature are compared and discussed in the context of structure-based ligand design. PMID- 7492543 TI - Screening of cyclic peptide phage libraries identifies ligands that bind streptavidin with high affinities. AB - The screening of combinatorial peptide libraries has emerged as an important tool in the discovery of novel substrates or ligands for enzyme and receptor targets. For example, screening linear peptide libraries using streptavidin as a model receptor system has previously identified many low-affinity peptide ligands, all of which contain the common motif His-Pro-Gln (HPQ). We reasoned that constraining the conformational freedom of linear peptides by cyclization in a library would yield peptide ligands of increased affinity. Three different cyclic peptide libraries were constructed in an M13 phage display system as N-terminal pIII protein fusions. The random peptide sequences were flanked by two cysteine residues, which allows efficient disulfide bond formation and cyclization during phage assembly. These cyclic peptide libraries were screened with streptavidin as the model receptor system. Many sequences, all of which contained the motif His Pro-Gln (HPQ), were discovered, and in the preceding paper, the structures of complexes of streptavidin-bound cyclic and linear peptides are described (Katz, 1995). Analysis of binding kinetics and affinities demonstrated that the conformationally constrained cyclic peptides bound streptavidin with affinities up to 3 orders of magnitude higher than linear peptides identified in previous library screens. These results demonstrate the potential of screening conformationally constrained peptide libraries for high-affinity novel receptor ligands or enzyme substrates. PMID- 7492544 TI - Isolation and characterization of rat cholangiocyte vesicles enriched in apical or basolateral plasma membrane domains. AB - Cholangiocytes, the epithelial cells that line intrahepatic bile ducts, are composed of plasma membranes with discrete apical (lumenal) and basolateral domains. While these domains are thought to contain different transporters, exchangers, channels, and receptors, no methodology currently exists for the isolation of these different membrane compartments. Thus, our aim was to develop a technique to isolated plasma membranes from cholangiocytes enriched in apical or basolateral domains. We isolate a cholangiocyte-enriched population of cells from rats 3 weeks after bile duct ligation (BDL), a maneuver which stimulates selective cholangiocyte proliferation. Using isopycnic centrifugation on linear sucrose gradients, we prepared a mixed cholangiocyte plasma membrane (MCPM) fraction from which we further generated separate apical and basolateral cholangiocyte plasma membrane (ACPM and BCPM, respectively). We characterized these fractions by specific marker enzyme assays, transmission electron microscopy (TEM), lipid analysis, anisotropy measurements, one- and two dimensional gel electrophoresis, and quantitative immunoblots of the cystic fibrosis transmembrane conductance regulator (CFTR). Marker enzyme assays and TEM revealed that the MCPM fraction was essentially devoid of other organelles but was enriched approximately 70-fold in phosphodiesterase I, a general plasma membrane marker; the ACPM and BCPM were appropriately enriched in the respective apical and basolateral markers. TEM of ACPM and BCPM revealed homogeneous preparations of vesiculated membranes without contamination by other organelles. Lipid analysis, one- and two-dimensional gel electrophoresis, CFTR immunoblots, and anisotropy measurements showed unequivocal differences in lipid and protein composition and in fluidity between the ACPM and BCPM domains.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7492545 TI - Allosteric modulation of acetylcholinesterase activity by peripheral ligands involves a conformational transition of the anionic subsite. AB - Replacement of residues Asp74, Trp286, and Tyr72, which are constituents of the peripheral anionic site (PAS) of human acetylcholinesterase (HuAChE), affected similarly both the binding and the inhibition constants of the PAS-specific ligand propidium, demonstrating that changes in the inhibitory activity are a direct consequence of altered binding to the PAS. In contrast, the active center HuAChE mutants W86A and Y133A show respective 350- and 25-fold increased resistance to inhibition by propidium but no change in binding affinities, demonstrating that the allosteric mechanism of PAS-mediated inhibition involves a conformational change of these Trp86 and Tyr133 residues rather than physical obstruction of substrate access by the inhibitor itself. These findings support the recent proposal that the allosteric mechanism operates via transition between active and nonactive conformations of the anionic subsite Trp86 and that replacement of Tyr133 by alanine may stabilize a nonactive Trp86 conformation that occludes the active center [Ordentlich et al. (1995) J. Biol. Chem. 270, 2082]. In further support of this mechanism and the role of Tyr133, we find that (a) the dissociation constants (Kd) for the noncovalent complexes of the irreversible inhibitors diisopropyl phosphorofluoridate or paraoxon with Y133A HuAChE are increased 20-500-fold, relative to either wild-type enzyme or its Y133F or W86A mutants; and (b) access of substrates such as 3,3-dimethylbutyl thioacetate is restored by removal of Trp86 from the Y133A enzyme (i.e., the W86A/Y133A mutant). We suggest that the conformational transition of Trp86 is coupled to the motions of the cysteine loop (Cys69-Cys96) of HuAChE and is inherent to the dynamics of the native enzyme. PMID- 7492548 TI - Comparison of secondary structures in water of chondroitin-4-sulfate and dermatan sulfate: implications in the formation of tertiary structures. AB - Chondroitin-4-sulfate (CS4) and dermatan sulfate (DS) were examined in D2O solution, with or without NaCl, by NMR at 300 MHz, to investigate the physicochemical consequences of epimerization of glucuronate (GlcUA in CS4) to iduronate (IdoUA in DS). Nuclear Overhauser effects (NOEs) and spin-lattice relaxation times following selective and nonselective inversion were measured at up to 70 degrees C. (1) NOEs confirmed 4C1 conformations of sugar rings in N acetylgalactosamine and GlcUA, and 1C4 or 2S0 in IdoUA. Conflict between NMR data and periodate oxidation kinetics over IdoUA conformations is resolvable by postulating conversion of monodentate periodate-1C4 complexes to conformations in which periodate oxidation can procede. (2) Pairs of glycosidic protons in CS4 and DS showed strong NOEs, implying that stretches of 2-fold helix were present, with carboxylate and acetamido groups close to each other on the same side of tapelike molecules, extending previous work in dimethyl sulfoxide solution. CS4 and DS have large hydrophobic patches in this configuration, similar to those in keratan sulfate and hyaluronan. (3) Selective and nonselective inversion-recoveries implied similar segmental and backbone mobilities and hence flexibilities in CS4 and DS. This is discussed in terms of intrinsic flexibility of glycosidic conformations, modified by hydrogen-bonded arrays. (4) We postulate that hydrophobic and hydrogen bonding drives DS self-aggregation. Stronger self aggregation of DS compared with CS4 is attributed to increased intermolecular hydrogen-bonding in DS, secondary to decreased intramolecular hydrogen-binding. This is partly because the axial OH groups in 1C4 IdoUA cannot hydrogen-bond to neighboring sugars as can the equatorial OH groups in GlcUA of CS4. PMID- 7492547 TI - alpha-Fluoro acid and alpha-fluoro amide analogs of acetyl-CoA as inhibitors of citrate synthase: effect of pKa matching on binding affinity and hydrogen bond length. AB - An alpha-fluoro acid analog and an alpha-fluoro amide analog of acetyl-CoA have been synthesized. The ternary complexes of these inhibitors with oxaloacetate and citrate synthase have been crystallized and their structures analyzed at 1.7 A resolution. The structures are similar to those reported for the corresponding non-fluorinated analogs (Usher et al., 1994), with all forming unusually short hydrogen bonds to Asp 375. The alpha-fluoro amide analog binds with an affinity 1.5-fold lower than that of a previously described amide analog lacking the alpha fluoro group. The alpha-fluoro acid analog binds with a 50-fold decreased affinity relative to the corresponding unfluorinated analog. The binding affinities are consistent with increased strengths of hydrogen bonds to Asp 375 with closer matching of pKa values between hydrogen bond donors and acceptors. The results do not support any direct correlation between hydrogen bond strength and hydrogen bond length in enzyme-inhibitor complexes. PMID- 7492549 TI - Anchoring of tryptophan and tyrosine analogs at the hydrocarbon-polar boundary in model membrane vesicles: parallax analysis of fluorescence quenching induced by nitroxide-labeled phospholipids. AB - The role of Trp and Tyr residues in determining membrane protein structure is particularly interesting because indole and phenol structures combine hydrophobic and polar groups, and it is hard to predict the exact region of the membrane at which their energy would be at a minimum. To determine the depths intrinsically favored by these residues, the locations of membrane-associating Trp and Tyr analogs have been determined using a fluorescence quenching technique able to measure depth at high resolution. They are found to locate at the same depths as Trp and Tyr in membrane proteins, 14-15 A from the bilayer center, which implies an important role for these residues in aligning membrane proteins in precise relationship to the lipid bilayer. PMID- 7492546 TI - An active site phenylalanine of 3-oxo-delta 5-steroid isomerase is catalytically important for proton transfer. AB - 3-oxo-delta 5-steroid isomerase (KSI) from Pseudomonas testosteroni catalyzes the isomerization of a variety of 3-oxo-delta 5-steroids to their conjugated delta 4 isomers through the intermediate formation of a dienolate ion. This dienolate is formed by proton transfer from C-4 of the substrate to Asp-38, which then protonates the dienolate at C-6. Catalysis is enhanced by electrophilic assistance (hydrogen bonding) to the 3-oxygen by Tyr-14. We have investigated the effect of modifying phenylalanine-101 (F101), a hydrophobic residue that is located in the binding pocket of KSI. Two mutant enzymes (F101L and F101A) of KSI were prepared, and their kinetic properties were examined with 5-androstene-3, 17 dione (1) as the substrate. Both of the mutants show reduced values of kcat compared to the wild type (WT), by about 30-fold (F101L) and by 270-fold (F101A), with only a small difference in Km values. There is little change in the Ki's ( < or = 4-fold) for the product 4-androstene-3,17-dione (3), although both enzymes bind the intermediate analog d-equilenin (4) about 25-fold less tightly than does the WT. Fluorescence spectra of 4 bound to each of these enzymes suggest that 4 is ionized at the active site of WT, un-ionized at the active site of F101A and a mixture of these ionization states at the active site of F101L. Free energy profiles are constructed for each of the mutant enzymes, and these are compared to the free energy profile for the WT.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7492550 TI - Crystal and molecular structure of a new Z-DNA crystal form: d[CGT(2-NH2-A)CG] and its platinated derivative. AB - The three-dimensional structure of d[CGTA'CG], where A' = [2-NH2-A], was determined to atomic (1.35 A) resolution by single isomorphous replacement. The d[CGTA'CG] hexamer crystallizes in space group P3221, and is not isomorphous with other DNA hexanucleotides. Despite completely different crystal packing, the essential characteristics of the Z-DNA conformation are maintained. The structure was determined by single isomorphous replacement using a triammine platinum fragment. Thus, this study also demonstrates, for the first time, the feasibility of the use of this reagent for the direct phasing of DNA crystal structures. PMID- 7492551 TI - Proton NMR studies of cytochrome c peroxidase mutant N82A: hyperfine resonance assignments, identification of two interconverting enzyme ofecies, quantitating the rate of interconversion, and determination of equilibrium constants. AB - The cyanide-ligated form of the baker's yeast cytochrome c peroxidase mutant bearing the mutation Asn82-->Ala82 ([N82A]CcPCN) has been studied by proton NMR spectroscopy. This mutation alters an amino acid that forms a hydrogen bond to His52, the distal histidine residue that interacts in the heme pocket with heme bound ligands. His52 is a residue critical to cytochrome c peroxidase's normal function. Proton hyperfine resonance assignments have been made for the cyanide ligated form of the mutant by comparison with 1-D and NOESY spectra of the wild type native enzyme. For [N82A]CcPCN, proton NMR spectra reveal two significant phenomena. First, similar to results published for the related mutant [N82D]CcPCN [Satterlee, J. D., et al. (1994) Eur. J. Biochem. 244, 81-87], for Ala82 mutation disrupts the hydrogen bond between His52 and the heme-ligated CN. Second, four of the 24 resolved hyperfine-shifted resonances are doubled in the mutant enzyme's proton spectrum, leading to the concept that the heme active site environment is dynamically microheterogeneous on a very localized scale. Two magnetically inequivalent enzyme forms are detected in a pure enzyme preparation. Varying temperature causes the two enzyme forms to interconvert. Magnetization transfer experiments further document this interconversion between enzyme forms and have been used to determine that the rate of interconversion is 250 (+/- 53) s-1. The equilibrium constant at 20 degrees C is 1.5. Equilibrium constants have been calculated at various temperatures between 5 and 29 degrees C leading to the following values: delta H = 60 kJ mol-1; delta S = 0.20 kJ K-1 mol-1. PMID- 7492552 TI - Reaction of cytochrome bo3 with oxygen: extra redox center(s) are present in the protein. AB - The reaction of oxygen with cytochrome bo3, a quinol oxidase from Escherichia coli, has been studied by resonance Raman scattering after initiation of the reaction by CO photolysis in a continuous flow apparatus and by directly mixing the enzyme with oxygen. The high-frequency region of the spectrum was monitored to determine the time evolution of the spin, oxidation, and coordination states of heme O and the oxidation state of heme B by using newly established marker lines for each heme. Three phases of the reaction were detected. In phase I, complete in 75 microseconds, O2 reacted with heme O and formed a low-spin ferric or ferryl adduct without significant oxidation of heme B. In phase II, between 75 and 120 microseconds, a small fraction of heme B was oxidized. In phase III, at approximately 1 s, the majority of heme B was oxidized and heme O reverted to a high-spin ferric state. The high rate of oxygen reduction at heme O to the three- or four-electron reduced level, despite a very low rate of heme B oxidation, indicates that there are electron donors active in the enzyme other than the metal centers. Assays of our enzyme preparations rule out a quinol in the tight binding (QH) site as a possible donor but instead suggest electron donation from the protein matrix, such as from tryptophans or tyrosine.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7492553 TI - Universality of energy and electron transfer processes in photosystem I. AB - Femtosecond transient absorption spectroscopy has been used to investigate the photoinduced energy and electron transfer processes in photosystem I (PS I) particles from cyanobacteria, green algae, and higher plants. At room temperature, the kinetics observed in all three species are very similar: Following 590 nm excitation, an equilibration process(es) with a 3.7-7.5 ps lifetime was observed, followed by a 19-24 ps process that is associated with trapping. In all three species long-wavelength pigments (pigments that absorb at longer wavelengths than the primary electron donor) were observed. The difference spectrum associated with reduction of the primary electron acceptor [Ao(-)-Ao) difference spectrum] was obtained for all three species. The (Ao(-)-Ao) difference spectra obtained from measurements using detergent-isolated PS I particles from spinach and Chlamydomonas reinhardtii are similar but clearly membrane fragments. In all three species the reduced primary electron acceptor (Ao(-)) is reoxidized extremely rapidly, in about 20 ps. The difference spectrum associated with Ao reduction appears to contain contributions from more than a single chlorophyll pigment. PMID- 7492554 TI - Kinetic analysis of glucose transporter trafficking in fibroblasts and adipocytes. AB - Insulin regulates hexose uptake by the redistribution of glucose transport proteins from intracellular compartments to the cell surface. We have submitted the trafficking of GLUT1, GLUT4, and GLUT1/GLUT4 chimeras to a mathematical analysis in the context of different models. Our data suggest that a model with one intracellular and one cell surface compartment can describe the glucose transporter-trafficking kinetics in fibroblasts. Moreover, the difference in cellular distribution between GLUT1 and GLUT4 overexpressed in fibroblasts is best explained by a slower rate of movement of GLUT4 to the plasma membrane. In 3T3-L1 adipocytes, glucose transporter-trafficking kinetics is adequately described by a three-pool model which includes flow of transporters from the endosomal compartment to cell surface. The kinetic roles of previously identified motifs in GLUT4 trafficking were defined in proposed fibroblast and adipocyte glucose transporter-trafficking models. The C-terminus is important in reducing the exocytosis rate from the endosomal compartment to the cell surface in both fibroblasts and adipocytes, and the N-terminus behaves similarly in adipocytes. The C-terminus has an additional signal(s) that allows GLUT4 to be sequestered more efficiently into the insulin responsive vesicle compartment. Mutation of the dileucine motif in the C-terminus significantly reduces the endocytosis of GLUT4 in both fibroblasts and adipocytes, but these amino acids appear not to be primarily responsible for the different kinetics of wild-type GLUT1 and GLUT4. PMID- 7492555 TI - Identification of a species specific regulatory site in human pancreatic cholesterol esterase. AB - All mammalian pancreatic cholesterol esterases (CEase) bind to membrane associated heparin at a single site on the intestinal brush border membrane with a dissociation constant of 100 nM. While the enzyme is bound to the membrane, the activity of the human and bovine enzymes is enhanced 2-fold when compared to the activity of the enzyme in solution. On the other hand, soluble heparin potently inhibits the human CEase-catalyzed hydrolysis of cholesterol oleate with an IC50 of 2 x 10(-4) mg/mL, a value that is about 10(4) times more potent than that found with the bovine enzyme. The C-terminal portion of the human enzyme contains 16 proline-rich repeats of 11 amino acids each, while that from other species contains only a few of these repeat units. To determine if the unique human C terminus is responsible for this inhibition, two chimeras containing either the human N-terminus (residues 1-445) and the bovine C-terminus (residues 446-557), HB, or the bovine N-terminus (residues 1-445) and the human C-terminus (residues 446-722), BH, were prepared. The cholesterol oleate hydrolytic activity of these chimeras was similar to that for the recombinant human and bovine enzymes. Importantly, each chimera was inhibited by heparin with IC50 values of 0.03 and 0.1 mg/mL for HB and BH, respectively. These intermediate IC50 values indicate that human CEase has two structural regions that contribute to is unique inhibition by this sulfated glycosaminoglycan, and these could regulate cholesterol uptake in humans. PMID- 7492556 TI - Mechanism-based inactivation of tRNA-guanine transglycosylase from Escherichia coli by 2-amino-5-(fluoromethyl)pyrrolo[2,3-d]pyrimidin-4 (3H)-one. AB - In Escherichia coli, tRNA-guanine transglycosylase (TGT) catalyzes the incorporation of the queuine precursor preQ1 [2-amino-5-(aminomethyl)pyrrolo[2,3 d]pyrimidin-4(3H)-one] into tRNA. This precursor is further elaborated to queuine by two subsequent enzymic reactions [Slany, R. K., & Kersten, H. (1994) Biochimie 76, 1178-1182]. Our previous studies [Hoops, G. C., Townsend, L. B., & Garcia, G. A., (1995) Biochemistry (in press)] on a series of synthetic 5- and 6-substituted 2-aminopyrrolo[2,3-d]pyrimidin-4(3H) -ones have revealed that the E. coli TGT tolerates a wide diversity of substituents (isosteric, or nearly so, to the aminomethyl group of preQ1) at the 5 position. We report here that 2-amino-5 (fluoromethyl)pyrrolo[2,3-d]pyrimidin-4 (3H)-one (FMPP) inactivates TGT in a time and concentration-dependent manner with k(inact) = 0.074 min-1 and KI = 136 microM. A competitive inhibitor (7-methyl-preQ1), with respect to preQ1, of TGT [Hoops, G.C., Townsend, L.B., & Garcia, G.A. (1995) Biochemistry (in press)] protects the enzyme from inactivation by FMPP. FMPP also acts as a competitive inhibitor (KI = 114 microM) of TGT under initial velocity conditions. The rate of fluoride release from FMPP is slightly faster (0.064 min-1) than the k(inact) (0.053 min-1) at 300 microM FMPP, consistent with fluoride release preceding inactivation. FMPP appears to partition between "normal" turnover (kcat = 0.461 min-1 and Km = 152 microM), inactivation, and an alternative processing to an unidentified, fluoride-released product.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7492557 TI - Effects of zinc finger mutations on the nucleic acid binding activities of Xenopus transcription factor IIIA. AB - Transcription factor IIIA (TFIIIA) is required for the activation of 5S RNA gene transcription as well as the storage of 5s RNA as a 7S ribonucleoprotein particle. Interaction with both nucleic acids is mediated through nine C2H2 zinc fingers. In order to determine amino acid regions necessary for nucleic acid interaction, a series of substitution mutants Xenopus laevis TFIIIA have been constructed and expressed as recombinant proteins in Escherichia coli. The mutant proteins were purified to homogeneity and analyzed for 5S RNA gene and 5S RNA binding activities using a nitrocellulose filter binding assay. All of the mutant TFIIIA proteins retained full 5S RNA binding activity. Substitution of fingers 2, 3, and 4-6 of TFIIIA with zinc finger sequences from other proteins significantly reduced the interaction of the protein with the 5S RNA gene. In contrast, substitution of finger 1 or finger 7 had little effect on the interaction of TFIIIA with the 5S RNA gene. The results of scanning substitution mutagenesis within the first three zinc fingers of TFIIIA suggested that DNA contacts made by the alpha-helical regions of finger 2 and particularly of finger 3 provide the majority of the free energy of the TFIIIA-DNA interaction. Basic amino acids found at the same position within the alpha-helices of fingers 2 and 3 of TFIIIA are required for high-affinity DNA binding activity. The identification of amino acid residues critical for the formation of a TFIIIA-DNA complex contributes to our understanding of zinc finger protein-nucleic acid interactions. PMID- 7492558 TI - X-ray crystallographic studies on hexameric insulins in the presence of helix stabilizing agents, thiocyanate, methylparaben, and phenol. AB - Three X-ray crystallographic studies have been carried out on pig insulin in the presence of three ligands, thiocyanate, methylparaben (methyl p-hydroxybenzoate), and phenol. In each case, rhombohedral crystals were obtained, which diffracted to 1.8, 1.9, and 2.3 A, respectively. Each crystal structure was very similar to that of 4-zinc pig insulin, which was used as a starting model for PROLSQ refinement (Collaborative Computational Project, Number 4, 1994). The R factors for the refined structures of thiocyanate insulin, methylparaben insulin, and phenol insulin were 19.6, 18.4, and 19.1, respectively. Each crystal structure consists of T3R3f insulin hexamers with two zinc ions per hexamer. In the R3f trimer of the thiocyanate insulin hexamer, one thiocyanate ion is coordinated to the zinc on the hexamer 3-fold axis, but there is no evidence of zinc ion binding in the off-axis zinc ion sites seen in the 4-zinc pig insulin structure. In the methylparaben insulin and phenol insulin hexamers, the phenolic ligands are bound at the dimer-dimer interfaces in the R3f trimers in a manner similar to that of phenol in R6 phenol insulin. The binding of methylparaben appears to make the hexamer more compact by drawing the A and the B chains closer together in the binding site. In all three structures presented herein, the conformations of the first three residues of the B chain in the R3f trimer are extended rather than alpha-helical, as is seen in R6 phenol insulin. The energetics of ligand binding in the insulin hexamer are discussed. PMID- 7492559 TI - Nuclear magnetic resonance studies demonstrate differences in the interaction of retinoic acid with two highly homologous cellular retinoic acid binding proteins. AB - Cellular retinoic acid binding protein-I (CRABP-I) and cellular retinoic acid binding protein-II (CRABP-II) are highly homologous, 15 kDa proteins which bind all-trans-retinoic acid. In the adult, CRABP-II is expressed predominately in the epidermis, while CRAPB-I is expressed in a variety of tissues. To obtain structural information which could aid the design of more selective ligands, isotope-directed NMR methods were employed to observe the CRABP-bound conformation of 13C-labeled retinoic acid and to identify its contact points with neighboring amino acids. Analysis of HMQC, HMQC-TOCSY, and 13C-TOCSY-REVINEPT on CRABP-bound (2,3,6,7,8,9,10,11,19-13C)- and (1,4,5,8,9,16, 17,18,19-13C)-all trans-retinoic acid allowed the unambiguous assignment of all labeled protons and their attached 13C resonances. The volumes of 16 olefinic proton-methyl NOE cross peaks measured from 30-ms 13C-(omega 2)-filtered 1H NOESY experiments were used to determine the conformations about the 6-, 8-, and 10-single bonds of the retinoic acid polyene chain. These spectra show qualitatively distinct NOE patterns for the two CRABPs. Measured cross-peak volumes for CRABP-II bound retinoic acid were well predicted by a single, static conformational having a 6-s torsion angle of -60 degrees skewed from a cis conformation. In contrast, for CRABP-I no single, static conformation was able to match the pattern of cross peaks, suggesting motion about the 6-s bond. The measured cross-peaks were best described by 8-s and 10-s torsion angles of 180 degrees +/- 30 degrees, a trans configuration, for both proteins. The pattern of intermolecular NOESY cross-peaks between 13C-labeled protons in the ring portion of retinoic acid and protein protons were different between CRABP-I and CRABP-II. These differences coincide well with nearby amino acid substitutions in the recently reported X-ray structures of crystalline CRABP-I and CRABP-II and may assist rational design of selective ligands. PMID- 7492560 TI - Large activation energy barriers to chaperone-peptide complex formation and dissociation. AB - To probe the mechanism of chaperone substrate selection, we have investigated the kinetics of complex formation and dissociation between the molecular chaperone DnaK and a short peptide (Cro, representing amino acids 1-12 of the cro repressor protein). The Cro protein was N-terminally labeled with the environmentally sensitive fluorophore dansyl chloride (Cro*), and steady-state and stopped-flow fluorescence spectroscopies and fluorescence-detected high-performance size exclusion chromatography (HPSEC) were used to monitor complex formation and dissociation over a range of temperatures in the absence of ATP. The results are summarized as follows: (i) Cro* binds to DnaK with a second-order rate constant, k(on), which varies from 8 to 200 M-1 s-1 between 15 and 37 degrees C. The slow on-rate is a consequence of a large activation energy barrier. The activation enthalpy (delta H*) and the prefactor [omega exp delta S*/R)] are 26 kcal mol-1 and 7 x 10(20) M-1 s-1, respectively. (ii) Once formed, DnaK-Cro* complexes are long-lived, especially at low temperatures (T < 15 degrees C). The off-rate is unusually temperature-sensitive, for example, there is a 478-fold increase in k(off) from 2.3 x 10(6) to 1.1 x 10(-3) s-1 over a range of only 30 degrees C (5 35 degrees C). The steep temperature-dependence of the off-rate is a consequence of a very large activation energy barrier to DnaK-Cro* complex dissociation [delta H* = 34.6 kcal mol-1 and omega exp (delta S*/R) = 2 x 10(21) s-1]. The relatively low affinity of the Cro* peptide for DnaK is due to a large kinetic barrier to binding. We discuss possible causes for these large kinetic barriers. PMID- 7492561 TI - Crystal structure of Ustilago sphaerogena ribonuclease U2 at 1.8 A resolution. AB - The crystal structure of purine-specific ribonuclease (RNase) U2 from Ustilago sphaerogena has been solved by the molecular replacement methods using RNase T1 as a search model. The structure, with 114 amino acid residues, 141 water molecules, and a sulfate ion, is refined to an R factor of 0.143 at 1.8 A resolution. As evidenced by the electron densities, residues 49 and 50 are revised to Glu 49 and Asp 50, respectively, and also Asp 45 is identified as a beta-isomerized form to L-isoaspartate with a beta-peptide linkage. RNase U2 consists of a beta-hairpin at residues from 7 to 14, a 4.4-turn alpha-helix from 16 to 32, a central beta-sheet with five strands, and a protruding beta-turn from 74 to 77. As for the catalytic site residues, His 41, Glu 62, and Arg 85 are located as constituents of the central beta-sheet, and Tyr 39 and His 101 are situated at either end of the beta-sheet. The side chains of Tyr 39, Glu 62, Arg 85, and His 101 are hydrogen-bonded to the sulfate ion which marks the RNA phosphate position. Though the side chain of His 41 is pointing away from the sulfate, small conformational adjustments of His 41 enable the side chain to interact with either the phosphate or the ribose group of RNA. The loop region from Tyr 44 to Asp 50 is ascribed to the base recognition site where Glu 49 is involved in adenine recognition. beta-Isomerized Asp 45 suggests that this region is conformationally flexible and alterable. PMID- 7492563 TI - Asp 46 can substitute Asp 96 as the Schiff base proton donor in bacteriorhodopsin. AB - Bacteriorhodopsin functions as a light-driven proton pump in the purple membrane of Halobacterium salinarium. A variety of studies have established that a proton is transferred over an approximately 10 A distance from Asp 96 to the retinylidene Schiff base during the M --> N transition of the bR photocycle. In order to further explore the mechanism of this Schiff base reprotonation, we compared the properties of the double mutant Thr 46 --> Asp/Asp 96 --> Asn (T46D/D96N), the single mutants Asp 96 --> Asn (D96N) and Thr 46 --> Asp (T46D), and wild-type bR. In contrast to D96N, which exhibits a very slow M decay, T46D/D96N has an M decay close to that of wild-type bR. FTIR difference spectroscopy detects bands in the carboxyl and carboxylate stretch region of T46D/D96N consistent with the deprotonation of Asp 46 during the M --> N transition. In addition, bands associated with structural changes of Asn 96 in the mutant D96N are absent in T46D/D96N. Resonance Raman spectroscopy provides evidence that both T46D/D96N and T46D have a long-lived N-like species in their photocycles. These data demonstrate that Asp 46 can substitute for Asp 96 as the proton donor group in the reprotonation pathway of the Schiff base during the M - > N transition. However, N decay is delayed in comparison to wild-type bR. This may be due to a partial block in the proton pathway leading from the cytoplasmic medium to Asp 46. PMID- 7492565 TI - Neonatal nursing in the Gaza Strip. PMID- 7492564 TI - Working in international war zones: a personal account. PMID- 7492562 TI - Kinetics of inorganic phosphate release during the interaction of p21ras with the GTPase-activating proteins, p120-GAP and neurofibromin. AB - The rate of GTP hydrolysis on p21ras is accelerated by approximately 10(5) times by the catalytic domains of GTPase-activating proteins (GAPs), p120-GAP (GAP-344) or neurofibromin (NF1-334). The kinetic mechanism of this activation has been investigated by following the release of inorganic phosphate (Pi), using a fluorescent probe that is sensitive to Pi [Brune, M., Hunter, J., Corrie, J. E. T., & Webb, M. R. (1994) Biochemistry 33, 8262-8271]. Measurements were made in real time with a stopped-flow apparatus, in which the p21ras complex with the 2',3'-methanthraniloyl analogue of GTP (mantGTP) was mixed with the GAP in the presence of this Pi probe. The results show that Pi release is fast and that the overall hydrolysis is controlled by the cleavage itself or a conformational change preceding the cleavage. The time courses were single exponentials over a range of [GAP-344] and were modeled to show that a single step controlled Pi release. The maximum rate constant was 15 s-1 (all data at 30 degrees C, pH 7.6, low ionic strength) in experiments in which GAP-344 underwent a single turnover, compared with 5 s-1 for multiple-turnover experiments, and possible causes of this discrepancy were investigated and discussed. With NF1-334 the time courses were more complex, showing a lag prior to rapid release of Pi. The results were consistent with a Kd of 0.04 microM for NF1-344 affinity is some 3 orders of magnitude tighter than that of GAP-344.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7492566 TI - The mobilization of nurses: experiences of a lifetime. Interview by Jane Harris. PMID- 7492567 TI - Signaling by the transforming growth factor-beta receptors. PMID- 7492568 TI - Mammalian nucleotide excision repair. PMID- 7492569 TI - Phosphoinositide-specific phospholipase C and mitogenic signaling. AB - The importance of PLC activation in cell proliferation is evident from the fact that the hydrolysis of PtdIns(4,5)P2 is one of the early events that follow the interaction of many growth factors and mitogens with their respective receptors. However, the importance of PLC activation is not restricted to proliferation; it is one of the most common transmembrane signaling events elicited by receptors that regulate many other cellular processes, including differentiation, metabolism, secretion, contraction, and sensory perception. It is also clear that cell proliferation signaling does not always require PLC, as indicated by the fact that growth factors such as insulin and CSF-1 do not appear to elicit the hydrolysis of PtdIns(4,5)P2, even though the intracellular domains of their receptors carry a PTK domain and the receptors show topologies very similar to those of the PLC-activating growth factors PDGF, EGF, and FGF. The growth factor dependent activation of PLC is initiated by the formation of a complex between the receptor PTK and PLC-gamma; the formation of this complex is mediated by a specific interaction between a tyrosine phosphate residue on the intracellular domain of PTK and the SH2 domain of PLC-gamma. The receptor PTK subsequently phosphorylates PLC-gamma, of which two distinct isozymes, PLC-gamma 1 and PLC gamma 2, have been identified. Proliferation of T cells and B cells in response to the aggregation of their respective cell surface receptors is also accompanied by the activation of PLC-gamma isozymes at an early stage. Unlike growth factor receptors, the T cell and B cell receptors lack intrinsic PTK activity but associate with several non-receptor PTKs of the Src and Syk families. Although the specific kinases are not known, one or more of these enzymes phosphorylate and activate PLC-gamma 1 and PLC-gamma 2. Transduction of growth signals by G protein-coupled receptors such as those for thrombin or bombesin also requires PtdIns(4,5)P2 hydrolysis, which, in this instance, is mediated by PLC-beta isozymes. The PLC-beta subfamily consists of four distinct members: PLC-beta 1, PLC-beta 2, PLC-beta 3, and PLC-beta 4. Agonist interaction with specific G protein-coupled receptors causes the dissociation of Gq proteins into G alpha and G beta gamma subunits and the exchange of GDP bound to G alpha for GTP. The resulting GTP-bound G alpha subunit then activates PLC-beta isoforms by binding to the carboxyl-terminal region of the enzyme.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7492570 TI - Polymorphonuclear leukocyte chemotaxis induced by zinc, copper and nickel in vitro. AB - Metallic dental restorations and prosthetic constructions are susceptible to corrosion in oral environment, resulting in the release of various heavy metal ions. Chloride salts of zinc, copper, nickel, chromium, iron and gold were tested for their ability to promote the migration of polymorphonuclear leukocytes (PMNs). Using a modified Boyden chamber assay for chemotaxis zinc, copper and nickel enhanced the migration of PMN cells in concentration range of 0.5-1.0 mM, whereas no augmentation in migratory activity was noted using chromium or iron. In contrast, an inhibition in migratory activity was observed in cells directed toward gold ions. Exposure of cells to zinc, copper or nickel ions induced an orientation reaction in leukocytes in a similar fashion as the polarization reaction induced by a potent peptide chemoattractant, N formylmethionylleucylphenylalanine (fMLP), in these cells. Exposure of PMN cells to zinc or nickel in chemotactic concentrations stimulated the chemotaxis of these cells to fMLP 2-fold, whereas pretreatment of the cells with zinc prior to assay markedly decreased the subsequent chemotactic migration of the cells to this metal or to fMLP. The enhanced locomotion of PMN cells induced by zinc, copper or nickel ions was found to be in greater extent due to an increase in directed migration (chemotaxis) rather than an augmentation in random movement (chemokinesis) as assessed by Zigmond-Hirsch checkerboard analysis. These results suggest that zinc, copper and nickel ions attract leukocytes by inducing and promoting the chemotactic response in these cells, which may modulate the inflammatory response of host tissue around such metals. PMID- 7492573 TI - Retinal Schiff base chromophore in the surfactant solubilised water pools in CCl4. AB - All-trans-N-retinylidene-n-butylamine Schiff base has been incorporated into AOT/CCl4 reverse micelles of various water pool sizes (omega = 0-20). The nature of interaction between retinal Schiff base and its surroundings in the micellar microenvironment has been investigated by NMR (1H-NMR and T1 studies). The Schiff base is found to undergo hydrogen bond interactions with bound water molecules. The results are discussed in terms of the implication of hydrogen bond interactions in retinal proteins, particularly, bacteriorhodopsin. PMID- 7492571 TI - Characterisation of the copper uptake mechanism and isolation of the ceruloplasmin receptor/copper transporter in human placental vesicles. AB - In this paper we have studied copper (Cu) uptake by microvillar vesicles isolated from human term placenta. We have characterised Cu uptake from CuHis2 complexes and shown that ceruloplasmin (Cp) inhibits uptake. Inhibition is complex and variable; in one series of experiments, the Vmax for uptake drops from 31.3 +/- 1.2 nmol/min per mg vesicle protein without added Cp to 11.3 +/- 1 nmol/min per mg vesicle protein at 91 micrograms/ml Cp. Similarly, the K0.5 increases from 0.35 +/- 0.08 microM to 1.35 +/- 0.25 microM, while the n value (the Hill coefficient) falls from 1.9 +/- 0.23 in the absence of Cp to 1.1 +/- 0.13 In another series, Cp had no effect below concentrations of about 100 micrograms/ml and in a third series only increased K0.5. The variability in effect seems to be related to the specific activity of the ceruloplasmin, which in turn is related to the copper complexes of the protein. The effect is specific for Cp; apotransferrin and a2-macroglobulin have no effect. 67Cu-labelled ceruloplasmin binds specifically to vesicles of term placenta with an affinity of 2.8 microU/mg vesicle protein and a Bmax of 79 microU/mg vesicle protein. CuHis2, but not histidine alone, can block the uptake. The data can be reconciled by proposing that the binding site of the transporter is relatively small and recognises a Cu dihistidine structure common to the low-molecular-weight complex and to the Type I and Type II coppers of ceruloplasmin. We have used these observations to develop an isolation method for the transporter and have identified it as a protein of M(r) 90,000 which is closely associated with alkaline phosphatase. There are also two proteins of M(r) 45,000 and 40,000 which may be breakdown products of the larger complex. Antibodies to the 45,000 protein block Cu binding and uptake from CuHis2 complexes, strongly implicating it as the copper transporter/ceruloplasmin receptor of human term placenta. PMID- 7492572 TI - Compared recognition of di- and trisulfide substrates by glutathione and trypanothione reductases. AB - Trypanothione trisulfide was synthesized according to two strategies. It was found to be recognized and reduced by trypanothione reductase as the natural disulfide substrate. At the difference with the mechanism observed for the reduction of glutathione trisulfide by glutathione reductase, the intermediate trypanothione persulfide was rapidly reduced. The enzymatic reduction of another trisulfide derived from an alternative substrate of trypanothione reductase was also studied. The structure of the trisulfide bridge of the substrate (intra- or intermolecular) appeared to be a determining factor in the enzymatic reduction pattern. Moreover, in the case of the alternative substrate of trypanothione reductase, differences of kinetics appeared for the first time between a di- and a trisulfide species. All kinetic parameters are given. PMID- 7492574 TI - Modulation by hypergravity of extracellular matrix macromolecules in in vitro human dermal fibroblasts. AB - In vitro human dermal fibroblasts were submitted to normal gravity (1 g) or to chronic hypergravity (20 g) over a period of 8 days. Changes in organization of extracellular matrix molecules were seen by indirect immunofluorescence. In the fibronectin layer, bundles of fibrils were gathered together leading to a disorganisation of the normal parallel pattern of fibers seen in control cultures. Type I collagen fibrils appeared with wooly outlines in controls whereas thick fibers were closely packed in 20-g cultures. A moderate increase of type III collagen fibril density was observed. No elastic fibers were seen in control or in 20-g cultures. In the culture medium, the release of soluble elastin (ELISA) and type I and III collagens (RIA) was undisturbed. Assays of enzymes involved in the remodeling of extracellular matrix showed an increase of cellular elastase activity (10%) and a decrease of the spontaneously active collagenase. Nevertheless, the total collagenase activity, (activated by trypsin), was increased by up to 30%. These data show a significant rise of the latent collagenase activity and suggest that release of the tissue inhibitor of metalloproteinase (TIMP1) was enhanced by hypergravity. PMID- 7492575 TI - The role of nitric oxide in hemodynamic and metabolic alterations induced by prostaglandin F2 alpha in the perfused rat liver. AB - In the liver prostaglandins have been shown to be potent regulators of portal blood flow, carbohydrate metabolism and bile secretion. It is not known whether these effects represent a direct action of prostaglandins, and it has been suggested that nitric oxide (NO) might be a critical mediator for prostaglandin induced hepatic events. We have studied whether nitric oxide formation or inhibition alters the action of prostaglandin F2 alpha (PG F2 alpha) in a single pass liver perfusion model. The liver of untreated rats (constitutive NO synthase) or after pretreatment with endotoxin (inducible form of NO-synthase) was perfused at a constant pressure via the portal vein. Effluate were collected in 1-min intervals and bile in 5-min intervals. In both groups the addition of PG F2 alpha (10 microM) to the perfusate for 5 min resulted in a significant increase of glucose and lactate production, and in a significant decrease in portal blood flow (-0.56 +/- 0.04 ml/g per min), in bile flow (-60.7%) and in bile acid release (-60.6%). Inhibition of NO synthase by adding NG-monomethyl-L arginine (L-NMMA, 100 microM) to the perfusate did not affect any of the alterations induced by PG F2 alpha. Substitution of the endogenous substrate for the NO synthase L-arginine (500 microM) in the perfusate completely prevented the hemodynamic alterations induced by PG F2 alpha in endotoxin pretreated livers and limited the flow reduction (0.15 +/- 0.04 ml/g per min) in the untreated group. The substitution of L-arginine in the perfusate of endotoxin pretreated livers raised nitrite (from 1.5 +/- 0.3 to 3.6 +/- 0.7 nmol/g per min) and urea release (from 65 +/- 25 to 294 +/- 68 nmol/g per min), but had no effect on any of the other metabolic parameters and bile secretion. We conclude that PG F2 alpha increases glucose and lactate production in the perfused rat liver and decreases portal flow bile secretion. The metabolic effects induced by PG F2 alpha appear to be independent of NO mediation and hemodynamic alterations. Portal flow alone can be influenced by endogenous NO formation. PMID- 7492576 TI - Family A cellulases: two essential tryptophan residues in endoglucanase III from Trichoderma reesei. AB - Three tryptophan residues are readily oxidised by N-bromosuccinimide in endoglucanase III from Trichoderma reesei. Evidence was obtained that the residue first modified is situated in the cellulose-binding domain and the second in the enzyme's catalytic site. The latter influences the binding and hydrolysis of soluble substrates. The modification of a third residue does not further affect the catalytic properties. The present results complement published data concerning other identified catalytic residues, and help to clarify the active site structure of family A cellulases. PMID- 7492577 TI - A novel 550-kDa protein in skeletal muscle of chick embryo: purification and localization. AB - We have found a novel protein with a molecular mass of 550 kDa on SDS polyacrylamide gels, which is abundant in skeletal muscle tissues at an early stage of chick embryonic development. The 550-kDa protein decreased with the progress of development, and only a slight amount of the protein was present in adult chicken skeletal muscle. The 550-kDa protein was purified from the cytoplasm of 18 day embryos by a procedure including ultracentrifugation and gel filtration. The purified 550-kDa protein was essentially free of contaminants as judged by SDS-PAGE. By immunofluorescence and immunoelectron microscopy using the antibody raised against the 550-kDa protein, this protein was shown to be localized in the peripheries of adult muscle fibers and at the Z-disks of isolated myofibrils. These findings have led us to conclude that the 550-kDa protein is a novel myofibrillar protein in chicken skeletal muscle. PMID- 7492578 TI - Characterization of a novel 550-kDa protein in skeletal muscle of chick embryo. AB - Some characteristics of a novel 550-kDa protein which is abundant in skeletal muscle tissues at an early stage of the chick embryo, and localized in the peripheries of adult muscle fibers and at the Z-disks of isolated myofibrils, was investigated. A cosedimentation experiment and solid phase immunoabsorbent assay showed that the 550-kDa protein binds directly to F-actin. Therefore, it is concluded that the 550-kDa protein is a novel actin-binding protein. The 550-kDa protein was also interacted with alpha-actinin, laminin, fibronectin and Type IV collagen. Reactions with several kinds of lectin revealed that the 550-kDa protein is a glycoprotein containing oligosaccharides. Electron microscopic observation of negatively stained 550-kDa protein showed that native 550-kDa protein molecules are particles with an average diameter of 26.5 nm, but those particles treated with ethanol/ether are filamentous structures. These results suggest that the 550-kDa protein in the cytoplasma of unorganized skeletal muscle tissues exists as lipid-protein complex. Consequently, the 550-kDa protein may play an important role in the binding of myofibrils to the basal lamina by interaction with F-actin, alpha-actinin, laminin, fibronectin or Type IV collagen. PMID- 7492579 TI - Polyamine metabolism in the rat liver after orthotopic liver transplantation. AB - We examined the polyamine metabolism in liver transplanted after cold ischemia and effects of putrescine administration on liver injury, liver regeneration, and survival rate after orthotopic liver transplantation in the rat. Male Wistar rats were used as donors and recipients. Grafts were stored in Euro-Collins solution for 6 h at 4 degrees C. Orthotopic liver transplantation was performed by the three cuff technique. The activities of aspartate aminotransferase, alanine aminotransferase and lactate dehydrogenase elevated and peaked 4 h after liver transplantation. Hepatic ornithine decarboxylase and spermidine/spermine N1 acetyltransferase activities were also elevated and peaked 8 h after the operation. In agreement with the increases in ornithine decarboxylase and spermidine/spermine N1-acetyltransferase activities, the putrescine content increased and spermidine content decreased in the transplanted liver. Putrescine administrated intraperitoneally improved the survival rate, decreased serum transaminase level and increased the [3H]thymidine incorporation into the liver DNA. These findings suggest that both biosynthetic and biodegradative pathways are stimulated in liver transplantation, resulting in the increase in the formation of putrescine from ornithine and from spermidine, and that putrescine administration improve the survival rate by protecting the damaged graft after cold ischemia and reperfusion and by stimulating liver regeneration. PMID- 7492580 TI - Enrichment of Penicillium chrysogenum microbodies by isopycnic centrifugation in nycodenz as visualized with immuno-electron microscopy. AB - A procedure to enrich microbodies from Penicillium chrysogenum and a method to evaluate the purity and integrity of the microbodies are described. As a P. chrysogenum microbody marker acyltransferase (AT) was used. The P. chrysogenum hyphae were converted into protoplasts with Novozym 234. In Percoll-sucrose buffer the protoplasts were separated from mycelial debris after 10,000 x g centrifugation. Purified protoplasts were lysed, and the cell homogenate was centrifuged to form a 14,000 x g pellet. After 2 h, 45,000 x g isopycnic centrifugation of the 14,000 x g pellet on a continuous 20-60% nycodenz gradient, ten fractions were collected. The fractions were analyzed for AT containing microbodies by immuno-blotting and immuno-electron microscopy. The results showed that AT-microbodies are enriched in the 38% nycodenz fraction. The microbodies had a diameter of 400 to 500 nm, revealed an intact single membrane and confined AT. The estimated equilibrium density of the P. chyrsogenum microbodies was 1.20 g ml-1 as deduced from the 38% (w/v) nycodenz concentration. PMID- 7492581 TI - Direct evidence for the elevated synthesis and secretion of procathepsin L in the distal caput epididymis of boar. AB - The proteins which are secreted from the restricted part of the epididymis are suggested to sustain sperm maturation. In porcine species, as the potential abilities of sperm for movement and fertilization greatly increase in the corpus epididymis, the secretions in both the caput and corpus epididymis seem to be very important for the sperm maturation. In this study, we have directed our attention to the 40 kDa protein which is detected in the fluid of the distal caput epididymis of boar. It was purified from the porcine cauda epididymal fluid and its cDNA was cloned from the cDNA library of the distal caput epididymis. According to the deduced amino acid sequence, the 40 kDa protein has been identified as procathepsin L. Northern blot analysis showed that the procathepsin L mRNA was most abundant in the distal caput epididymis among the tissues as examined. Consistent with the distribution of the procathepsin L mRNA in the epididymis, the activity of procathepsin L was absent in the fluid of the proximal and mid caput epididymis and first appeared in the distal caput epididymal fluid, whose contents gradually decreased with the passage through the epididymis. These results first appeared in the first distal caput epididymis expresses very high levels of procathepsin L and unusually secretes it into the luminal fluid instead of targeting it to lysosomes. It has been also found that the mRNA of PDGF, which is known to enhance cathepsin L expression in the culture cells, is very high in the mid caput epididymis, which just precedes the site of procathepsin L secretion. This result indicates that PDGF directly regulates the locally restricted expression and secretion of procathepsin L in the epididymis, which is one of the possible mechanisms involved in the functional differentiation in the epididymis. PMID- 7492582 TI - Conformation of the propeptide domain of factor IX. AB - The propeptide domain in the precursor forms of blood clotting proteins contains the recognition sequences for gamma-carboxylase. In hemophilia B, several point mutations in this propeptide domain are responsible for the inherited disease. A peptide containing the propeptide sequence of factor IX was synthesized by solid phase methods. Two dimensional 1H-NMR and CD studies indicate that this peptide motif adopts an alpha-helical structure in a 40% trifluoroethanol-containing aqueous solution. The results suggest that the amphipathic alpha-helix within the propeptide domain of factor IX could create a recognition surface for gamma carboxylase. The influences of mutations and their relationship with the alpha helical structure are discussed. PMID- 7492583 TI - Isolation, characterization and mode of neutralization of a potent antihemorrhagic factor from the serum of the snake Bothrops asper. AB - A potent antihemorrhagic factor (BaSAH1) was isolated from the serum of the snake Bothrops asper by ammonium sulfate precipitation at 40-60%, Sephacryl S-200 and Sephadex G-50 gel filtration, DEAE-Sepharose, and hydrophobic Phenyl-Sepharose chromatography. The purified protein showed one band with an isoelectric point of 5.2 and a molecular weight of 66 kDa. 4 micrograms of the purified factor BaSAH were needed to neutralize the hemorrhagic dose of B. asper whole venom compared to 60 micrograms of the clinically used horse polyvalent immunoglobulins. Moreover, 0.35 microgram of BaSAH were sufficient to achieve complete neutralization of the main hemorrhagic toxin (BaH1), with a molar ratio of 2:1. The antihemorrhagic activity was stable between pH 1.5-9 and up to 60 degrees C but lost activity completely after 30 min of heating at 70 degrees C. BaSAH did not digest the hemorrhagic toxin BaH1 or formed a precipitin line with it, nor with the whole venom. Both ELISA experiments and chromatography of BaSAH after incubation with the 125I-labeled hemorrhagic toxin BaH1 demonstrated that the mechanism of the neutralization involves a formation of an inactive soluble complex between the natural antihemorrhagin and the main hemorrhagin of B. asper venom. PMID- 7492584 TI - Partial purification and characterization of histidine acetyltransferase in brain of Nile tilapia (Oreochromis niloticus). AB - High activity of histidine acetyltransferase (HISAT) was found in the brain and the lens of Nile tilapia Oreochromis niloticus. HISAT was semi-purified 4166-fold from the brain of Nile tilapia. The affinity chromatography using a Blue Sepharose 6 FF column was very effective for purification of this enzyme. The enzyme had a broad pH optimum from pH 7.0 to pH 9.5, and did not require any divalent metal ion. The semi-purified HISAT showed a strict substrate specificity for L-histidine (and its methyl derivatives) and acetylcoenzyme A (CoASAc). The reaction velocity fits normal Michaelis-Menten kinetics with respect to both L histidine (Km, 0.45 mM) and CoASAc (Km, 0.027 mM). Gel filtration on Superdex 200 HR indicated the molecular weight of 39,000. It was presumed that the 38.5 kDa protein, which was intensely visualized in sodium dodecyl sulfate-polyacrylamide gel electrophoresis, was a single subunit derived from HISAT. PMID- 7492585 TI - Molecular activation-deactivation of xanthine oxidase in human milk. AB - Enzymic activity and protein levels of xanthine oxidase were measured in serial samples of breast milk donated by each of 14 mothers, starting, in all but two cases, within 7 days following parturition. Enzyme activity varied widely, usually reaching peak values during the first 15 days and falling thereafter, by as much as 98%, to basal levels that were subsequently largely maintained. Corresponding changes in xanthine oxidase protein levels were not observed and, consequently, the specific activity of xanthine oxidase followed the above pattern. The capacity of human xanthine oxidase to undergo activation deactivation cycles at the molecular level has important implications, not only for its role in breast milk, but also for its potential as a source of reactive oxygen species in other human tissues. PMID- 7492586 TI - Iron- and peroxide-dependent conjugation of dopamine with cysteine: oxidative routes to the novel brain metabolite 5-S-cysteinyldopamine. AB - The mechanism of formation of 5-S-cysteinyldopamine (5-S-CDA), a putative index of oxidative stress in dopaminergic regions of the brain, was investigated by comparing the ability of a number of neurochemically relevant oxidising systems to promote the conjugation of dopamine with cysteine in vitro. Autoxidation of the catecholamine proceeds at relatively slow rate in the physiological pH range, and is little affected by 1 mM Fe(2+)-EDTA complex. In the presence of cysteine, however, the Fe(2+)-induced autoxidation is hastened, affording little amounts of 5-S-CDA. Formation of the adduct is completely suppressed by ascorbic acid. Hydrogen peroxide, in the presence of Fe(2+)-EDTA (Fenton-type oxidation) or peroxidase, promotes a relatively efficient conversion of dopamine to 5-S-CDA and the minor isomer 2-S-CDA. Noteworthy, 15-hydroperoxyeicosatetraenoic acid (arachidonic acid hydroperoxide, HPETE), in the presence of Fe(2+)-EDTA complex, can also mediate 5-S-CDA formation, whilst superoxide radicals are little effective. Overall, these results suggest that ferrous ions, hydrogen peroxide and lipoperoxides may play an important role in 5-S-CDA generation. PMID- 7492587 TI - In vivo 19F-NMR spectroscopic study of halothane uptake in rabbit brain. AB - Uptake of a fluorinated anesthetic, halothane, in rabbit brain and blood was studied using 19F-NMR spectroscopic techniques. Localized one-dimensional chemical shift imaging and non-localized one-pulse sequence were used to measure brain uptake kinetics in vivo. Halothane signal was found predominantly in the cerebral cortex. Uptake in the brain followed a first-order biexponential kinetics. The average half-lives were 4 min and 70 min, respectively, for the 'fast' and 'slow' phases of the uptake. Uptake in the arterial blood was also biexponential. However, equilibration of halothane in the brain considerably lagged behind that in arterial blood. This delay was ascribed to a 'restricted diffusion' of the anesthetic molecule into brain tissue. PMID- 7492589 TI - Resonance Raman spectroscopy reveals novel ligation properties of the porcine myoglobin double mutant H64V/V68H. AB - A resonance Raman spectroscopic study of the porcine myoglobin double mutant H64V/V68H has confirmed that the ferric form is bis-histidine ligated, has revealed that the bis-histidine ligation is retained on reduction to the ferrous form, and has demonstrated that CO can displace the ligated distal histidine to produce a ferrous CO form which has a low steady-state photolability, indicating that the replacement histidine blocks the CO escape route from the binding site. PMID- 7492590 TI - Specific dinucleoside polyphosphate cleaving enzymes from chromaffin cells: a fluorimetric study. AB - This article presents a fluorimetric study of the main properties of the enzymes dinucleoside tetraphosphate (asymmetrical) hydrolase or dinucleoside tetraphosphatase (Ap4Aase, EC 3.6.1.17) and dinucleoside triphosphate hydrolase or dinucleoside triphosphatase (Ap3Aase, EC 3.6.1.29), both present in adrenal medulla cytosolic extracts. Diethenoadenosine polyphosphates, epsilon-(ApnA), are used as artificial fluorogenic substrates. Ap4Aase exhibits a molecular mass around 20 kDa and neutral optimum pH (7.0-7.5). It requires Mg2+ and preferentially hydrolyzes substrates with four phosphate groups. Km for epsilon (Ap4A) is 1.3 microM and Ki for Ap4A and Gp4G are 1 and 0.2 microM respectively. Km for Ap4A determined by HPLC is 1.6 microM. epsilon-(Ap5A) and epsilon-(Ap6A) are hydrolyzed at reduced rates. This enzyme is inhibited by Zn2+, F- and very strongly by Ap4 and epsilon-Ap4. Ca2+ cannot replace Mg2+, but behaves as inhibitor in its presence. The substrate analogs dinucleoside triphosphates Ap3A, G;3G, m7Gp3G and m7Gp3A and the periodate-oxidized nucleotides o-(Ap4A), o epsilon-(Ap4A), o-Ap4 and o epsilon-Ap4 behave as inhibitors. Ap3Aase exhibits a molecular mass around 30 kDa and neutral optimum pH (7.0-7.5). It requires Mg2+ or Ca2+, but retains a low measurable activity around 10% in the absence of these divalent cations. It only hydrolyzes substrates with three phosphate groups. Km for epsilon-(Ap3A) is 11 microM and Ki for Ap3A and Gp3G are 20 and 22 microM, respectively. Km for Ap3A determined by HPLC is 16 microM. m7Gp3G and m7Gp3A are also good substrates for triphosphatase. PMID- 7492591 TI - Stereochemical course of glucan hydrolysis by barley (1-->3)- and (1-->3, 1-->4) beta-glucanases. AB - The stereochemical course of hydrolysis of Laminaria digitata laminarin and barley (1-->3, 1-->4)-beta-glucan by barley (1-->3)-beta-glucanase (E.C. 3.2.1.39) isoenzyme GII and (1-->3, 1-->4)-beta-glucanase (EC 3.2.1.73) isoenzyme EII, respectively, has been determined by 1H-NMR. Both enzymes catalyse hydrolysis with retention of anomeric configuration (e-->e) and may therefore operate via a double displacement mechanism. We predict that all other members of Family 17 of beta-glycosyl hydrolases also follow this stereochemical course of hydrolysis. PMID- 7492588 TI - Hampered expression of isoaspartyl protein carboxyl methyltransferase gene in the human cataractous lens. AB - Isoaspartyl protein carboxyl methyltransferase (PIMT) is implicated in the repair of age-damaged proteins by converting altered aspartic acid residues to normal L aspartic acid residues. Northern blot and reverse transcription (RT)-PCR analyses have revealed that PIMT gene expression in the human lens is detected exclusively in epithelial cells, and that the mRNA levels in cataractous lens epithelia are significantly lower than those in normal age-matched lens tissue. These results suggest that PIMT may play a vital role in maintaining the clarity of the lens and preventing cataract formation. PMID- 7492592 TI - Mechanism-based inactivation of porcine kidney diamine oxidase by 1,4-diamino-2 butene. AB - Cis- and trans-1,4-diamino-2-butene are substrates and potent inactivators of porcine kidney diamine oxidase. Evidence from absorption and NMR spectra indicates that both are oxidized to pyrrole. Both substrates are irreversible mechanism-based inactivators of the enzyme, although the trans isomer is more potent and results in complete inactivation in a reaction which follows pseudo first-order kinetics with an apparent Ki of 0.34 mM and a second-order inactivation constant of 500 M-1 s-1. Under the same conditions, 46% of the activity remains when the enzyme is reacted with cis-1,4-diamino-2-butene. Trans 4-amino-2-butenal, the product of oxidation of the trans diamine, has been synthesized and shown to undergo cyclization to pyrrole in a concentration dependent manner, approaching second-order at low concentrations. Trans-4-amino-2 butenal is itself a potent irreversible inhibitor with IC50 of 2.5 microM. We propose that the irreversible inactivation by both cis- and trans-1,4-diamino-2 butene involves attack by a protein-based nucleophilic residue on the unsaturated aminoenal products of the enzymatic reactions, resulting in a covalent adduct. Cyclization of the cis-aminoenal to pyrrole is much more rapid than in the trans case, thus it is less available for inhibitory reaction with the protein. PMID- 7492593 TI - The topology of CuA in relation to the other metal centres in cytochrome-c oxidase of Saccharomyces cerevisiae as determined by analysis of second-site reversions. AB - Second-site revertants were selected from a respiratory-deficient mutant carrying the mutation D369N located in a loop between helices IX and X close to H376 and H378, the proposed ligands of haem a3 and haem a, respectively. A reversion was observed in subunit II, in the vicinity of the CuA ligands. This same reversion compensates the subunit I deficiency mutation, S140L, assumed to be near H62, the second putative histidine ligand to haem a. These data enable us to propose a three-dimensional topology in which CuA in subunit II is located on top of the Positive-side of subunit I and in proximity to all three of its metal centres. PMID- 7492594 TI - Bovine pancreatic ribonuclease A as a model of an enzyme with multiple substrate binding sites. AB - Bovine pancreatic ribonuclease A is an enzyme that catalyses the depolymerization of RNA. This process involves the interaction of the enzyme with the polymeric substrate in the active site and its correct alignment on the surface of the enzyme through multiple binding subsites that essentially recognize the negatively charged phosphate groups of the substrate. The enzyme shows a strong specificity for pyrimidine bases at the 3'-position of the phosphodiester bond that is cleaved and a preference for purine bases at the 5'-position and, probably, for guanine at the next position. On the other hand, the enzyme shows a clear preference for polynucleotide substrates over oligonucleotides. In this review the contributions to the catalytic mechanism of some amino-acid residues that are located at non catalytic binding subsites are analysed. PMID- 7492595 TI - The ATPase activity of purified CDC48p from Saccharomyces cerevisiae shows complex dependence on ATP-, ADP-, and NADH-concentrations and is completely inhibited by NEM. AB - The cell cycle protein CDC48p from Saccharomyces cerevisiae is a member of a protein superfamily (AAA superfamily) characterized by a common region of approximately 200 amino-acid residues including an ATP binding consensus. CDC48p purified to homogeneity showed considerable ATPase activity which could be completely abolished by preincubation with NEM in the absence of ATP. ATP protects the protein from NEM and stabilizes the otherwise labile enzyme. The ATPase activity is reversibly inhibited by NADH and shows cooperativity with its substrate ATP. The application of the in vitro ATPase activity to the identification of physiologically interacting molecules is discussed. By electron microscopy, the enzyme was shown to consist of hexameric ring structures similar to its vertebrate homologue. PMID- 7492596 TI - A precursor form of human kidney gamma-glutamyl transferase in normal and cancerous tissues, and its possible post-translational modification. AB - We have found three molecular forms of human gamma-glutamyl transferase (GGT) in normal and renal cell carcinomatous tissues, and also have reported the marked differences in the sugar chains of GGTs between normal and cancerous tissues by serial lectin affinity chromatographies. In this study, the peptide maps of three purified GGTs (79 kDa, 50 kDa and 25 kDa) obtained by lysylendopeptidase digestion, the subcellular localization of GGTs, and the sugar chains of GGTs were compared between normal and cancerous tissues. According to the results, the total peptide bands of the digested 79 kDa component represented the sum of those of the digested 50 and 25 kDa components on 12.5% SDS-PAGE. In addition, C terminal and N-terminal amino-acid sequences of the 79 kDa protein were the same as the sequences of light and heavy subunits, respectively, suggesting that the 79 kDa component is of the precursor form of the 50 kDa mature heavy and 25 kDa light subunits, respectively. On the other hand, the GGT activity in renal cell carcinomatous tissues was significantly increased in the microsomal fraction and decreased in the soluble fraction compared with that of normal tissues. Meanwhile, the sugar moiety of GGTs in the respective subcellular fractions was obviously different between normal and cancerous tissues. In particular, a reduced multiantennary complex type sugar chain and an elevated high-mannose or hybrid-type sugar chain in the microsomal fraction were observed in the GGT in cancerous tissues. PMID- 7492597 TI - Sucrose reduces the efficiency of protein denaturation by a chaotropic agent. AB - Sugars and polyols are used to stabilize proteins. The degree of stabilization conferred on a model protein by sucrose was calculated in terms of the free energy of folding. Phosphoglycerate kinase (PGK) was denatured by guanidine hydrochloride (GuHCl) in different sucrose concentrations. The linear extrapolation method [1,2] was used to calculate the free energy of folding in the absence of denaturant. Although sucrose increased the concentration of GuHCl required to unfold the protein, the free energy of folding in water was unchanged. In order to probe the nature of the stabilizing effect of sucrose, an FT-Raman spectroscopic study of denaturant-polyol systems was undertaken. Investigations of interactions between GuHCl, urea or formamide and polyhydric compounds, revealed no evidence for hydrogen bonding or dipole-dipole associations. Polyhydric compounds caused minor changes in denaturant spectra although the converse was not observed. The structure of deuterated water changed on addition of denaturants. For non-ionic denaturants, addition of polyhydric solutes countered this change in water structure. Thus polyhydric compounds oppose the effect of denaturants on water structure. The observed increase in GuHCl concentration required to unfold PGK in the presence of sucrose may be attributed to this property of sucrose. PMID- 7492598 TI - Inactivation and conformation changes of the glycated and non-glycated D glyceraldehyde-3-phosphate dehydrogenase during guanidine-HCl denaturation. AB - The glycated D-glyceraldehyde-3-phosphate dehydrogenases have been isolated from rabbit muscle and erythrocytes (He et al. (1995) Biochem. J. 309, 133-139). The circular dichroism spectrum in the near-ultraviolet of gGAPDH was different from that of GAPDH. Changes in intrinsic protein fluorescence and in the 410 nm fluorescence of the NAD derivatives introduced at the active sites of both the glycated and non-glycated GAPDH from rabbit were compared on inactivation during denaturation in GuHCl. Complete inactivation for the non-glycated enzyme occurred in 0.5 M GuHCl solution, however, that for the glycated enzyme occurred in the 0.35 M solution. The kinetic inactivation of gGAPDH was a biphasic process (the fast and slow phases). The fast phase for gGAPDH was faster than that of GAPDH. The kinetic exposure of the fluorescent NAD derivatives at the active sites of both enzymes was also biphasic with fast phase rates which approach those of the inactivation. It appears that glycation of the enzyme may disturb the spatial geometry of the functional groups responsible for the catalytic mechanism and affect the activity. PMID- 7492599 TI - Hydroxyquinones are competitive non-peptide inhibitors of HIV-1 proteinase. AB - Quinones with one, two and three aromatic rings are a new class of micromolar non peptidic inhibitors of HIV-1 proteinase, an enzyme essential for replication of Human Immunodeficiency Virus and an important drug target for AIDS. Substituted anthraquinones bearing hydroxyl substituents on one of their three rings were the most potent of these inhibitors. Comparisons with other small non-peptidic inhibitors that are now emerging, together with enzyme kinetic data indicating that alizarin is a competitive inhibitor, suggest that anthraquinones bind in the active-site groove of HIV-1 proteinase. PMID- 7492600 TI - Antigenicity of topochemically related peptides. AB - Antibodies raised in rabbits against multimeric all-L peptides (MAP's) were first made monospecific by affinity chromatography on immobilized antigen columns and then tested for their ability to cross-react with topologically related variants of the parent antigen, where the chirality of each amino-acid residue (inverso derivatives), or the peptide sequence orientation (retro derivatives), was inverted, or where both modifications were simultaneously introduced (retro inverso derivatives). Retro, inverso, and retro-inverso forms of the parent peptide were prepared, both in the linear as well as in the BSA-conjugated form, and found to cross-react to a significant extent with affinity purified polyclonal antibodies raised against the parent peptide. Peptide variants displayed similar dose-dependent inhibitory effects on the interaction between immobilized parent antigen and affinity purified antibodies. Analysis of molecular models of the peptide variants in the trans-configuration suggested that the topological equivalence of alternating side chains in the series of related peptides may be responsible for the observed cross-recognition, leading to the formation of similar recognition surfaces which could mimic the parent peptide antigenic structure. PMID- 7492601 TI - Spin-labeling probe on conformational change at the active sites of creatine kinase during denaturation by guanidine hydrochloride. AB - The conformational change at the active sites of creatine kinase and its protection by substrates during guanidine denaturation were investigated by monitoring the ESR spectra of the nitroxide radical covalently bound to the reactive thiols of the enzyme. For the enzyme undenatured (pH 9.0) and in the presence of low concentrations of guanidine, i.e. less than 1 M, there are two kinds of enzyme molecule, one of which is bearing a compact structure at the active site and the other is of a looser structure. The content of the latter increases with increasing denaturant concentration. At concentrations of guanidine hydrochloride higher than 1 M, the structure of the enzyme molecule is monomorphic and becomes looser and looser with an increase of guanidine hydrochloride concentration. The existence of a nucleotide substrate complex protects the structure at the active sites of the enzyme from being changed, up to a concentration of denaturant of 0.2 M, while creatine has no protective effect. PMID- 7492602 TI - Fourier-transform infrared spectroscopic investigation of protein stability in the lyophilized form. AB - Upon the removal of water, proteins undergo a major, reversible rearrangement of their secondary structure, as revealed by FTIR spectroscopy. We have found herein that for recombinant human albumin (rHA) the extent of this structural change does not depend significantly either on the composition of the aqueous solution prior to lyophilization (protein concentration, pH, and the presence of excipients such as dextran or NaCl) or on the mode of dehydration (lyophilization, spray drying, or rotary evaporation), even though these factors profoundly affect rHA's solid-state stability against moisture-induced aggregation. In all cases, the alpha-helix content of rHA drops from 58% in solution to 25-35% in the dehydrated state, the beta-sheet content rises from 0 to 10-20%, and unordered structures increase from 40% to 50-60%. We have also investigated another model protein, hen egg-white lysozyme, and confirmed that it too undergoes a significant alteration of the secondary structure upon lyophilization. The extent of this structural reorganization has been found to be insensitive to the pH of the aqueous solution prior to lyophilization from pH 1.9 to 5.1, even though the thermal transition temperature (Tm) in aqueous solution over this range varies by 30 degrees C. PMID- 7492603 TI - Protein modification by the degradation products of ascorbate: formation of a novel pyrrole from the Maillard reaction of L-threose with proteins. AB - Ascorbate (vitamin C) degradation products can undergo non-enzymatic glycation (Maillard reaction) with proteins to form highly crosslinked structures with brown pigmentation and characteristic fluorescence. Proteins in the body, especially the long-lived proteins develop similar changes during aging and diabetes. Several studies have shown excessive degradation of ascorbate in plasma in diabetes, and in ocular lens during aging and cataract formation. Recent studies have suggested that ascorbate degradation products-mediated glycation plays a role in lens pigmentation and cataract formation. However, the precise chemical nature of ascorbate-specific advanced glycation end-products are not known. Here, we report the purification and characterization of a glycation end product derived from one of the major degradation products of ascorbate, L threose. This compound was characterized to be 2-acetamido-6-(3-(1,2 dihydroxyethyl)-2-formyl-4-hydroxymethyl-1- pyrrolyl)hexanoic acid (formyl threosyl pyrrole or FTP) formed by the condensation of epsilon-amino group of lysine with two molecules of threose. Formation of FTP occurred rapidly in the incubation of threose and lysine and reached plateau level within a day. We have developed a sensitive assay for its quantification in proteins based on enzyme digestion followed by HPLC. Ribonuclease A and human lens crystallins incubated with L-threose showed time- and sugar concentration-dependent increases in FTP, reaching 8.2 and 2.48 nmol per mg protein, respectively after one week of incubation. Human plasma proteins showed a peak with identical retention time as that of purified FTP under two different HPLC conditions. FTP may be used as a sensitive marker to assess ascorbate-mediated protein glycation and modifications in aging and diabetes. PMID- 7492604 TI - Kinetics of butyrylcholinesterase in reversed micelles under high pressure. AB - The combined effects of high pressure and reversed micelles have been studied to modulate the catalytic behaviour of butyrylcholinesterase. The purpose of this study was to determine whether the conformational plasticity of the enzyme is altered by entrapment in reversed micelles. The presence of soman, an irreversible inhibitor of cholinesterase was used to bring to the fore a possible modification of the enzyme behaviour in this system under pressure. Results show differences between enzyme in conventional medium and in reversed micelles regarding the mechanism of BuChE catalyzed hydrolysis of acetylthiocholine. In both systems, the enzyme displays a non-Michaelian behaviour with this substrate. In conventional medium the kinetics is multiphasic with an activation phase followed by an inhibition phase at high concentration. In reversed micelles there is inhibition by excess substrate but the activation phase is missing. This behaviour may be the result of a change of the enzyme conformational plasticity when is entrapped in reversed micelles. PMID- 7492605 TI - Processing and activation of recombinant mouse mastocytoma histidine decarboxylase in the particulate fraction of Sf9 cells by porcine pancreatic elastase. AB - Mature 53 kDa histidine decarboxylase (HDC) peptide is produced from a precursor 74 kDa peptide. The mechanism of specific cleavage by processing enzyme is unknown. Using the recombinant mouse 74 kDa HDC, we found that porcine pancreatic elastase specifically converted the inactive 74 kDa HDC to its active form of 53 kDa HDC. PMID- 7492607 TI - Inhibition of lipoxygenase in lentil protoplasts by expression of antisense RNA. AB - A number of plasmids were constructed containing chimeric genes consisting of fragments of antisense-oriented lentil lipoxygenase cDNA. The different constructs were tested for their ability to lower lipoxygenase activity in lentil protoplasts. Plasmids containing a full length lentil lipoxygenase cDNA proved to be the most effective, reducing the activity of the target enzyme to 70% of the control value. On the other hand, the full length lentil lipoxygenase cDNA in the sense orientation yielded a 20% increase of lipoxygenase activity. PMID- 7492606 TI - Effects of mixed solvents on three elementary steps in the reactions of horseradish peroxidase and lactoperoxidase. AB - The effects of methanol, acetone, and ethylene glycol (up to 50% v/v) on elementary steps in the reactions of horseradish peroxidase (HRP) and lactoperoxidase (LPO) were studied by means of the stopped-flow method and the difference spectrum. The rate constant (k3,app) of the oxidation reaction of p cresol with HRP compound II was remarkably reduced in the presence of organic solvents (to 2.3%, 1.8% and 9.4% of the original value in the presence of 50% (v/v) of methanol, acetone and ethylene glycol, respectively), then to a lesser degree were decreased the rate of the oxidation reaction with LPO compound II, and the rate of the oxidation reaction with HRP compound I. These reductions in the reaction rates were not due to competitive inhibition of the solvents, but considered to be related to the degree of exposure of the electron transfer route to the medium. While the rate constant of compound I formation (k1,app) was moderately affected by organic solvents in the case of HRP, the reaction rate with LPO was scarcely affected by organic solvents, being in harmony with the compact heme crevice which probably hampers penetration of solvent molecules. The rate constant (k2,i,app) of the oxidation reaction of an iodide ion by HRP compound I was also hardly affected by the solvents. On the basis of these facts, the mechanism of electron transfer from donors to compound I and compound II is discussed. PMID- 7492609 TI - Molecular mechanisms of diverse actions of prostanoid receptors. AB - This review summarizes recent advances in the molecular characterization of prostanoid receptors. Prostanoids exert versatile actions in diverse tissues and cells through specific cell surface receptors. Molecular biological studies revealed the primary structure of eight types and subtypes of prostanoid receptor from various species. These include the thromboxane A2 receptor, prostacyclin receptor, prostaglandin (PG) F receptor, PGD receptor and four subtypes of PGE receptors. They are coupled to different signal transduction systems. In addition, multiple isoforms of PGE receptor EP3 subtype have been identified in various species. They are produced through alternative RNA splicing from a single gene and differ only in their carboxy-terminal tails. These isoforms differ in the efficiency of G protein activation, in the specificity of coupling to G proteins or in sensitivity to desensitization. This molecular characterization is useful for understanding the diverse physiological roles of prostanoids. PMID- 7492608 TI - Compound D609 inhibits phorbol ester-stimulated phospholipase D activity and phospholipase C-mediated phosphatidylethanolamine hydrolysis. AB - Tricyclodecan-9-yl-xanthogenate (compound D609) has recently been used in various cellular systems to specifically inhibit the activity of phosphatidylcholine (PtdCho)-directed phospholipase C (PLC). Here we show that in intact NIH 3T3 fibroblasts, concentrations of D609 (35 to 50 micrograms/ml) which have been used to inhibit PLC activity also significantly inhibit phorbol ester-induced phospholipase D-mediated hydrolysis of both PtdCho and phosphatidylethanolamine (PtdEtn). In addition, in isolated membranes compound D609 also inhibited PLC mediated PtdEtn hydrolysis. The results indicate that compound D609 cannot be considered as a specific inhibitor of PtdCho-directed PLC activity. PMID- 7492610 TI - Identification and purification of a novel phospholipid/ganglioside-binding protein in rabbit serum. AB - We have isolated a novel phospholipid/ganglioside-binding protein from rabbit sera or platelet-free plasma. Using an affinity chromatography of a commercial gel (Sephacryl S-series gel, Pharmacia) column and a preparative polyacrylamide gel electrophoresis, the protein can be easily purified. The protein agglutinates human red cells irrespective of the ABO blood types, and its hemagglutination reaction is specifically inhibited by some phospholipids (phosphatidylserine and phosphatidylglycerol) and ganglioside (N-acetylneuraminyl-galactosylglucosyl ceramide, GM3). The hemagglutination and its inhibition reactions are independent on any divalent cations (Ca2+, Mg2+, Mn2+, Ni2+). The protein seems to be assembled as multimers of disulfide-bonded molecular of 86 kDa and 59 kDa subunits. PMID- 7492611 TI - Cholesterol precipitation from cholesterol-supersaturated bile models. AB - Bile-model systems containing cholesterol (CH), phosphatidylcholine (PC) and sodium cholate (NaC) at concentrations similar to those found in supersaturated human gall bladder bile ([CH]/[PC] = 0.60 +/- 0.01; CH saturation index, CSI = 1.58 +/- 0.03) were prepared by mixing PC-CH vesicles with NaC micellar solutions. Following mixing, the dispersion became transparent and gave rise to high resolution 1H-NMR spectra typical of mixed micellar systems. Cryo transmission electron micrographs of specimens vitrified at that stage support the conclusion that the vesicles had become completely micellized. Following micellization, the metastable (cholesterol-supersaturated) bile-models spontaneously underwent a series of reorganizational steps: first, cholesterol rich vesicles with a [CH]/[PC] ratio of 1.57 +/- 0.69 were formed, in co existence with a mixed micellar system with [CH]/[PC] = 0.43 +/- 0.01 and CSI = 1.12 +/- 0.03. The resultant cholesterol-rich vesicles subsequently aggregated and cholesterol crystals of varying sizes and shapes appeared within the aggregates: needle-like structures were first observed, followed by clusters of those crystals and of helical crystals. Eventually, typical plate-like cholesterol crystals appeared, at which time some of the PC returned to the non particulate (isotropic) phase. Consequently, the system contained cholesterol crystals coexisting with mixed micelles, whose composition was close to the limit of saturation (CSI = 1.08). These findings confirm the sequence of events preceding the appearance of cholesterol crystals, as previously proposed in our less detailed studies ((1990) Hepatology 12, 149S) and support the relevance of the morphologically similar results of Konikoff et al. ((1992) J. Clin. Invest. 90, 1155) obtained in a very dilute supersaturated bile-model. PMID- 7492613 TI - Cholesterol biosynthesis from lanosterol: regulation and purification of rat hepatic sterol 14-reductase. AB - We have previously characterized the membrane-bound sterol 14-reductase (14 reductase) that catalyzes anaerobically NADPH-dependent reduction of the 14 double bond of delta 8,14-diene or delta 7,14-diene sterols that are sterol intermediates in cholesterol biosynthesis in mammals (Paik et al. (1984) J. Biol. Chem. 259, 13413-13423). To elucidate the regulatory mechanism as well as molecular characteristics of the 14-reductase, we extended our investigation on the consequences of alteration of the enzymic activity under various physiological conditions. The enzymic activity of rat hepatic sterol 14-reductase was induced more than 11-fold by feeding 5% cholestyramine plus 0.1% lovastatin (the CL-diet) for 7 days but was severely suppressed by feeding 5% cholesterol or 0.01% AY-9944 (an inhibitor of 14-reductase) for the same period. The increase or decrease in the 14-reductase activity also parallels the same change in the cholesterol synthetic rate in hepatocytes from rats that had been fed either the CL-diet or 0.01% AY-9944. In vitro inhibition studies revealed that AY-9944 acts as a competitive inhibitor of the 14-reductase (Ki = 0.26 microM). A diurnal variation was observed for the 14-reductase with peak activity near the middle of the dark cycle (10 p.m.), which was abolished by administration of cycloheximide. With induced enzyme conditions 14-reductase has been further purified with chromatographic procedures to near homogeneity. Purified 14-reductase appears to be a M(r) = 70,000 protein that is composed of two equally-sized subunits having a M(r) = 38,000. All properties of the purified 14-reductase suggest that the solubilized enzyme is the principal 14-reductase of microsomes. Taken together, our results provide the first evidence in support of a previously unknown regulatory role for the 14-reductase in the overall cholesterol synthetic pathway. PMID- 7492612 TI - Inhibition of metal-catalyzed oxidation of low-density lipoprotein by free and albumin-bound bilirubin. AB - Both free and albumin-bound bilirubin are known to scavenge peroxyl radicals in vitro. In the present work we showed that free and albumin-bound bilirubin at the physiological concentration of the bile pigment in blood plasma could greatly inhibit the metal-catalyzed oxidation of low density lipoprotein (LDL) as shown by the reduced thiobarbituric acid reactivity, smaller or no shifts in electrophoretic mobility, less apo B fragmentation and a decreased amount of cholesterol oxidation products as detected by gas chromatography. Free bilirubin (BR) was more effective in inhibiting the production of thiobarbituric acid reactive substances in iron-catalyzed LDL peroxidation as compared to the copper catalyzed reaction up to a BR to metal molar ratio of 4:1. Above this ratio the same degree of inhibition was observed for both metal ions. It was found that serum albumin provided full protection against Cu(2+)-dependent oxidative stress only at very high protein to metal molar ratio, i.e., 30:1, that is similar to that in human plasma. Complexation of BR to albumin brought about a marked increase in the capacity of the complex to bind metal ions, particularly iron, as opposed to albumin alone. At a molar ratio of metal ion to albumin-BR of 1:1 the inhibition of lipid peroxidation was about 96% and it was almost complete at a molar ratio of 1:2. The ability of albumin-BR complex to inhibit effectively the transition metals-dependent oxidative stress could be important in the extravascular space where local concentrations of metal ions may exceed the protein binding capacity. In addition, the strong binding of iron to the albumin BR complex may be clinically important, especially in iron loaded sera of hemochromatosis patients, where the transferrin is fully saturated with this ion and the free iron could catalyze lipid peroxidation unless bound by a metal trapping device such as the albumin-BR complex. PMID- 7492614 TI - Genomic and cDNA cloning of a novel mouse lipoxygenase gene. AB - A novel 12- and 15-lipoxygenase related gene was isolated from a mouse strain 129 genomic phage library in a screen with a human 15-lipoxygenase cDNA probe. The complete genomic sequence revealed 14 exons and 13 introns covering 7.3 kb of DNA. The splice junctions were verified from the cDNA sequences of a series of overlapping RT-PCR products. This novel mouse lipoxygenase gene was found to contain an open reading frame encoding 662 amino acids. The cDNA sequence is 68% identical to the mouse leukocyte type 12-lipoxygenase and 65% identical to the mouse platelet type 12-lipoxygenase cDNA. Nucleotide identities to the human 15 lipoxygenase cDNA and the human platelet type 12-lipoxygenase cDNA are 70% and 67%, respectively. The deduced amino acid sequence is 60% identical to the mouse platelet and leukocyte type 12-lipoxygenase sequences and the human 15- and 12 lipoxygenase sequences. This novel mouse lipoxygenase gene was found to be transcribed predominantly in epidermal tissue and named Aloxe. PMID- 7492615 TI - Alkaline sphingomyelinase activity in rat gastrointestinal tract: distribution and characteristics. AB - Previous studies indicated that there was an alkaline sphingomyelinase (SMase) activity in small intestine, but its properties have not been studied in detail. In the present work, we studied the distribution of this enzyme activity in rat gastrointestinal tract and characterized it in intestinal mucosal homogenates. Little alkaline SMase activity was detected in the stomach and the duodenum. The activity in both mucosa and intestinal content increased in the small intestine and reached the maximum at the distal jejunum, then declined in the ileum and slightly increased again in the colon. The activity distribution pattern differed markedly from those of acid SMase and alkaline phosphatase. Little alkaline SMase activity could be found in bile, liver and pancreas before or after treatment with trypsin. The optimum pH of the alkaline SMase was 9. It specifically hydrolyzed sphingomyelin (SM), not phosphatidylcholine, to ceramide and phosphocholine. The alkaline SMase was bile salt dependent and was optionally activated by 3 mM bile salts. Triton X-100 could not mimic the effect of bile salt, rather dose-dependently inhibited the enzyme activity. Ca2+, Mg2+ did not change the alkaline SMase activity in the presence of bile salts, and reduced the activity in the absence of bile salt. Trypsin inactivated acid SMase in pancreas, liver and duodenum but had no influence on intestinal alkaline SMase activity. In conclusion, the intestinal alkaline SMase has a specific distribution pattern and the characters of it differ in several respects from the known acid and neutral SMases. PMID- 7492618 TI - Preparative-scale kinetic resolutions catalysed by microbial lipases immobilised in AOT-stabilised microemulsion-based organogels: cryoenzymology as a tool for improving enantioselectivity. AB - Gelatin-containing microemulsion based organogels have been used as an immobilisation matrix for lipases from a number of different sources. Kinetic resolutions of octan-3-ol, 1-octen-3-ol and 1-octyn-3-ol by esterification with decanoic acid have been performed using Chromobacterium viscosum (CV) lipase. CV lipase is highly enantioselective in favour of the (R)-(-) isomer of octan-3-ol, but the enantioselectivity is both reversed and decreased by the introduction of unsaturation at the 1-position. Marked improvements in enantioselectivity were achieved by carrying out the reaction at -15 degrees C, the enantiomeric excess of the ester product increasing from 47% (E = 3) to 73% (E = 8) in the case of 1 octen-3-ol, and from 17% (E = 1.4) to 38% (E = 2.5) in the case of 1-octyn-3-ol. The enantiomeric excess was approximately 85% (E approximately 15) for octan-3 ol, and there was no marked improvement in enantioselectivity even at -15 degrees C. Apparent activation energies for the esterification using decanoic acid of octan-3-ol, 1-octen-3-ol and 1-octyn-3-ol by CV lipase were 32 kJ mol-1, 31 kJ mol-1 and 41 kJ mol-1, respectively. This compares to an activation energy of 21 kJ mol-1 for the esterification of octan-1-ol with decanoic acid using CV lipase under the same conditions. Lipases from Pseudomonas (Fluka), Pseudomonas (Genzyme) and lipoprotein lipase ex Microbial (Genzyme) also selectively esterified the (R)-(-) isomer of racemic octan-3-ol, the two Pseudomonas preparations yielding product with an enantiomeric excess of 90%. Candida cylindracea lipase did not exhibit activity in gelatin-containing MBGs. Large scale syntheses were performed in a 1 dm3 batch reactor in which 200 cm3 of pelleted MBG (containing 350 mg of CV lipase) was used repeatedly for the kinetic resolution of octan-3-ol. PMID- 7492617 TI - Inhibitory effects of cerulenin on protein palmitoylation and insulin internalization in rat adipocytes. AB - Protein acylation by long-chain fatty acids has been suggested as a necessary step in membrane trafficking. Because several insulin effects are dependent upon membrane trafficking, the cellular effects of the protein acylation inhibitor cerulenin were examined. Cerulenin blocked palmitoylation of selected rat adipocyte proteins including CD36, the dominant marker for palmitoylation in adipocytes. To measure cerulenin's effects on insulin internalization, rat adipocytes were incubated with 125I-insulin at 37 degrees C in the presence or absence of cerulenin. Surface-bound and intracellular insulin were discriminated by the sensitivity of the former to rapid dissociation by a pH 3 buffer at 4 degrees C. Insulin internalization was inhibited 85% by 0.3 mM cerulenin. Inhibition required preincubation with the agent, was irreversible, was not dependent upon protein synthesis, and was not the result of ATP depletion. Cerulenin was also found to inhibit insulin-stimulated glucose uptake and acetyl CoA carboxylase activity. Cerulenin had no effect on basal glucose uptake and utilization or on the uptake and retention of fatty acids. In summary, protein acylation may be an important step in insulin-regulated cellular functions dependent upon membrane trafficking, such as insulin internalization. PMID- 7492616 TI - Phosphonate analogues of triacylglycerols are potent inhibitors of lipase. AB - 1,2-Dioctylcarbamoylglycero-3-O-p-nitrophenyl alkylphosphonates, with alkyl being methyl or octyl, were synthesised and tested as irreversible inhibitors of cutinase from Fusarium solani pisi and Staphylococcus hyicus lipase. Rapid inactivation of these enzymes occurred with a concomitant release of one mole of p-nitrophenol per mole of enzyme. With both lipases a higher reactivity was observed when the alkyl substituent on the phosphonate is a methyl rather than an octyl chain. Both lipases are highly selective for the chirality of these compounds at glycerol and at phosphorus. Rapid inactivation at an inhibitor concentration of 0.1 mol% in 100 mM NaTDOC (t 1/2 < 60 min.) occurred when the glycerol moiety had the (R) configuration, while inhibitors of the (S) configuration react 4-10-fold more slowly. The isomer with the p-nitrophenyl octylphosphonate attached to the secondary hydroxyl group of glycerol hardly inhibited (t 1/2 > 1 day) the lipases. These results reflect the known positional and stereopreference of these enzymes which preferentially release the fatty acid at sn-3 of natural triacylglycerols. The enzymes appeared to be even more selective for the chirality at phosphorus, the differences in reactivity of the faster and slower reacting isomers being as high as about 250-fold for the methylphosphonates and about 60-fold for the octylphosphonates. These phosphonates can be regarded as true active site-directed inhibitors. The inhibited enzymes can be considered as analogues of the tetrahedral intermediate in the acylation step that occurs during triacylglycerol hydrolysis. PMID- 7492619 TI - Studies on the metabolism of [1-14C]5.8.11-eicosatrienoic (Mead) acid in rat hepatocytes. AB - The oxidation, esterification and formation of chain elongated and desaturated products of [1-14C]5,8,11-eicosatrienoic (Mead) acid was studied. Liver cells from essentially fatty acid deficient (EFAD) and control rats were used. The metabolism of [1-14C]20:4, n-6 and [1-14C]20:5, n-3 were studied under the same experimental conditions. More 20:3, n-9 than 20:4, n-6 and 20:5, n-3 was oxidised both in EFAD and control cells. 20:3, n-9 was elongated to [14C]22:3, n-9 in both cell types and significant amounts of [14C]22:4, n-9 were formed in EFAD cells. Less 20:3, n-9 was esterified in phospholipids and more in triacylglycerol than observed with 20:4, n-6 and 20:5, n-3 in both cell types. 20:3, n-9 was mainly esterified in phosphatidylcholine and little was esterified in phosphatidylethanolamine compared to 20:4, n-6 and 20:5, n-3. In comparison, 20:3, n-9 was rather efficiently esterified in phosphatidylinositol as 18:0-20:3. [14C]22:4, n-9 formed from 20:3, n-9 in EFAD hepatocytes was esterified in triacylglycerol, not in phospholipids, unlike [14C]22:5, n-6 and [14C]22:6, n-3 which were mainly esterified in phospholipids. PMID- 7492620 TI - Electron paramagnetic resonance investigation of cytochrome P-450c21 from bovine adrenocortical microsomes: a new enzymatic activity. AB - Cytochrome P-450c21 (CYP21A1) purified from bovine adrenocortical microsomes was investigated by electron paramagnetic resonance (EPR) spectroscopy to clarify the interactions among heme active center, protein surroundings, water molecules and bound substrates or analogues. The low-spin EPR signals of the oxidized enzyme (as purified) consisted of two species; one at gz = 2.39, gy = 2.23, and gx = 1.925 (component A) and the other at gz = 2.42, gy = 2.23, and gx = 1.916 (component B). The component A is probably representing a product-bound form, whereas the component B indicates either occupation of the substrate-binding site with a substrate analogue or absence of steroid at the site. Upon addition of progesterone, the component A signal completely disappeared and the intensity of high-spin signal (g = 8.06, 3.54) increased. Addition of 17 alpha hydroxyprogesterone caused a development of a new low-spin signal at gz = 2.42, gy = 2.21, and gx = 1.966 (component C) and a further increase in intensity of the high-spin signal (g = 8.06, 3.54). Addition of 20 beta-hydroxyprogesterone caused an increase in intensity of the component C signal (and the g = 8 high spin signal) even stronger than did 17 alpha-hydroxyprogesterone. These observations suggest that 20 beta-hydroxyprogesterone binds to the cytochrome P 450c21 active center in a very similar manner as 17 alpha-hydroxyprogesterone does and, therefore, may be a metabolizable substrate. A new enzymatic pathway catalyzed by cytochrome P-450c21 was confirmed with a reconstituted enzymatic system consisting of cytochrome P-450c21, NADPH-cytochrome P-450 reductase and an NADPH-generating system. 20 beta-Hydroxyprogesterone was converted to progesterone via a putative 20 beta-oxidase reaction in a comparable turnover number to that of the 21-hydroxylation of 17 alpha-hydroxyprogesterone. PMID- 7492621 TI - Biochemical properties of cloned lipases from the Pseudomonas family. AB - Three Pseudomonas lipases, representing three subfamilies, were analysed for pH optima, destabilization by EGTA and surfactants, phospholipase and cholesterolesterase side activities. All the Pseudomonas lipases tested showed alkaline pH optima. The Pseudomonas cepacia and the P. pseudoalcaligenes lipases were totally inhibited by EGTA at pH 9, and the latter was also fully inhibited at pH 7. The lipase from P. mendocina was not inhibited by EGTA at any of the pH values tested. These findings indicate that a calcium binding site exists in some of the Pseudomonas lipases. The P. pseudoalcaligenes, P. cepacia and P. mendocina lipases were inhibited by the anionic surfactant SDS at concentrations between 0.01-0.5 mg/ml. The P. pseudoalcaligenes and P. cepacia lipases were not inhibited by the nonionic surfactant Brij35 in concentration up to 1 mg/ml, whereas the lipase from P. mendocina was inhibited at 0.1 mg/ml. The P. pseudoalcaligenes and P. cepacia lipases were found to possess high cholesterol esterase activity. P. pseudoalcaligenes lipase was further found to have high phospholipase activity. Ten Pseudomonas lipase sequences were compared by automatic sequence alignment. On the basis of sequence identity we have classified Pseudomonas lipases into five subfamilies. PMID- 7492622 TI - High dose of fluvastatin sodium (XU62-320), a new inhibitor of 3-hydroxy-3 methylglutaryl coenzyme A reductase, lowers plasma cholesterol levels in homozygous Watanabe-heritable hyperlipidemic rabbits. AB - The effects of fluvastatin sodium (XU62-320), a new type of inhibitor of 3 hydroxy-3-methylglutaryl coenzyme A reductase, on plasma cholesterol and triacylglycerol levels were investigated using homozygous Watanabe-heritable hyperlipidemic (WHHL) rabbit, an LDL-receptor-deficient animal which expresses a hepatic LDL receptor activity less than 5% that of control rabbits. Plasma levels of total, VLDL- and LDL-cholesterol were decreased profoundly after oral administration of fluvastatin at a dose of 50 mg/kg per day for 4 weeks. Plasma triacylglycerol levels were not affected by fluvastatin. Hepatic HMG-CoA reductase activity increased by 3-fold and hepatic LDL receptor activity increased by only 3.7-fold, as calculated by Scatchard plot analysis, with fluvastatin administration for 4 weeks, and the hepatic mRNA level for the rabbit LDL receptor was increased by 3-fold. Combined administration of fluvastatin (50 mg/kg per day) and cholestyramine, a bile acid sequestrant resin, at a level of 2% of the diet for 4 weeks more profoundly decreased plasma total, VLDL- and LDL cholesterol levels with induction of hepatic cholesterol 7 alpha-hydroxylase and no further induction of the hepatic LDL receptor. Plasma triacylglycerol levels were increased by the combination treatment. These results suggest that high dose of fluvastatin sodium is effective in lowering plasma cholesterol levels in homozygous WHHL rabbits through the shared mechanisms involving decrease in production and secretion of cholesterol from the liver and the induction of hepatic LDL receptor. Additional effect of cholestyramine on decrease in plasma cholesterol levels seems to be due to the further decrease in hepatic cholesterol secretion by up-regulation of hepatic cholesterol 7 alpha-hydroxylase. PMID- 7492623 TI - Graft-versus-leukemia and graft-versus-lymphoma effects of allogeneic bone marrow transplantation and of allogeneic donor leukocyte transfusions. PMID- 7492624 TI - High-dose cytosine arabinoside and daunorubicin postremission therapy in adults with de novo acute myeloid leukemia. Long-term follow-up of a prospective multicenter trial. AB - A total of 149 consecutive de novo AML patients aged 50 years or less (median age = 37 years) were enrolled in this prospective multicenter trial initiated in May 1985. All patients received the same induction and early consolidation therapy with daunorubicin (DNR), cytosine arabinoside (Ara-C), and etoposide (DAV). High dose Ara-C/DNR therapy included Ara-C at 3 g/m2, in 12 doses (HD-Ara-C/DNR I) and eight doses (HD-Ara-C/DNR II), followed by DNR 30 mg/m2 for 3 days. A complete remission (CR) was achieved in 104 (70%) patients; 61 complete responders received at least one cycle with HD-Ara-C/DNR. If those patients who were transplanted in first CR (n = 26), were not considered, the median relapse-free survival (MRFS) of the remaining 78 patients was 15 months, with a probability of relapse-free survival (RFS) at 116 months of 30% (95% CI, 20-40%) after a median follow-up of 95 months. The MRFS of the HD-Ara-C/DNR consolidated patients was 25 months, with a probability of RFS at 116 months of 37% (95% CI, 24-50%). If all patients who were transplanted (n = 44) were not considered, the median survival time (MST) was 18 months with a probability of being alive at 118 months of 24% (95% CI, 16-33%). MST of the HD-Ara-C/DNR consolidated patients was 58 months with a survival probability of 46% (95% CI, 31-60%) at 118 months. Prognostic factor analysis did not reveal any significant influence of age, sex, FAB subtype, white blood cell count, hemoglobin level, thrombocyte count, LDH, or response to the first induction course on RFS of the HD-Ara-C/DNR consolidated patients. In summary, HD-Ara-C/DNR consolidation can improve the long-term outcome of a subgroup of de novo AML patients. Further improvement of the outcome seems to depend on the identification of patients with an inferior outcome under that strategy who might benefit from alternative treatment strategies. PMID- 7492625 TI - Autografting with blood progenitor cells: predictive value of preapheresis blood cell counts on progenitor cell harvest and correlation of the reinfused cell dose with hematopoietic reconstitution. AB - One hundred and nine patients suffering from various malignancies underwent 285 apheresis procedures for PBPC collection. A median of two leukaphereses (range: 2 5) resulted in median numbers of 4.6 x 10(8) MNC/kg, 14.1 x 10(4) CFU-GM/kg, and 6.0 x 10(6) CD34+ cells/kg. Preleukapheresis peripheral blood CD34+ cells correlated significantly with collected CD34+ cells/kg (r = 0.94; p < 0.0001) and with CFU-GM/kg (r = 0.52; p < 0.0001). A value > 4 x 10(4) CD34+ cells/ml was highly predictive for a collection yield > 2.5 x 10(6) CD34+ cells/kg harvested by a single leukapheresis. Sixty patients were evaluated for hematologic reconstitution and engrafted in a median time of 10 days for WBC > 1.0 x 10(9)/l (range: 7-21 days), 10 days for ANC > 0.5 x 10(9)/l (7-20) and 11 days for PLT > 20 x 10(9)/l (7-62). Reinfused CD34+ cells/kg correlated significantly with hematologic engraftment (r = 0.44-0.52 and p < 0.006-0.001) as well as CFU-GM/kg (r = 0.36-0.44 and p < 0.007-0.001). A progenitor cell dose > 2.5 x 10(6) CD34+ cells/kg or > 8.0 x 10(4) CFU-GM/kg led to a significantly faster recovery for WBC, ANC, and PLT when compared with patients receiving < 2.5 x 10(6) CD34+ cells/kg or < 8.0 x 10(4) CFU-GM/kg. We conclude that rapid hematopoietic engraftment after high-dose therapy and PBPC reinfusion correlates well with a progenitor cell dose > 2.5 x 10(6) CD34+ cells/kg or > 8.0 x 10(4) CFU-GM/kg, and that above a preleukapheresis threshold of 4 x 10(4) CD34+ cells/ml a PBPC autograft containing > 2.5 x 10(6) CD34+ cells/kg can be collected by a single leukapheresis. We suggest that patients recovering from myelosuppression should be monitored for CD34+ cells in serial blood samples to determine the course of circulating hematopoietic progenitor cells. This issue will help to define the optimal time point to start apheresis and to predict a PBPC autograft harvested by a single leukapheresis, which will lead to rapid and stable hematopoietic reconstitution following transplantation. PMID- 7492626 TI - Human chemokines: enhancement of specific activity and effects in vitro on normal and leukemic progenitors and a factor-dependent cell line and in vivo in mice. AB - The myelosuppressive effects of human chemokines were evaluated in vitro on normal myeloid progenitors obtained from bone marrow and cord blood, on bone marrow progenitors from patients with acute or chronic leukemia, on proliferation of human factor-dependent cell line M07e, and in vivo on myelopoiesis in mice. Preincubation of human MIP-1 alpha, MIP-2 alpha, interleukin (IL)-8, platelet factor (PF) 4, monocyte chemotactic and activating factor (MCAF), and interferon inducible protein-10 (IP-10) in an acetonitrile (ACN) solution significantly enhanced the specific activity of these chemokines for in vitro suppression of granulocyte-macrophage (CFU-GM), erythroid (BFU-E), and multipotential (CFU-GEMM) progenitor cells stimulated to proliferate with a colony stimulating factor plus steel factor (SLF). Combinations of any two of these ACN-treated chemokines synergized to suppress colony formation of CFU-GM, BFU-E, and CFU-GEMM at chemokine concentrations below that at which combinations of non-ACN treated chemokines are active. Cord blood progenitors, as previously reported, were in a slow or noncycling state and nonresponsive to inhibition by chemokines. However, after suspension culture with GM-CSF, IL-3, and SLF, they were placed into rapid cell cycle and were responsive to inhibition by ACN-treated chemokines. Low doses of these ACN-pretreated chemokines were active in vivo in suppressing absolute numbers and cycling status of femoral marrow CFU-GM, BFU-E, and CFU-GEMM in C3H/HeJ mice. Other chemokines, alone and in combination, including MIP-1 beta, MIP-2 beta, GRO-alpha NAP-2, and RANTES, were inactive in vitro and in vivo whether or not they were pretreated with ACN. While heterogeneity in responsiveness of CFU-GM from different patients with leukemia to suppression by ACN-treated chemokines was apparent, if the patients had CFU-GM responsive to one of the active chemokines these cells were responsive to the other active chemokines; if patient CFU-GM were not responsive to one of the chemokines, they were not responsive to the other active chemokines. M07e colony-forming cells were responsive to the growth-inhibiting effects of the active ACN-treated chemokines, alone and in combination, but these effects were rapidly reversible and sustained only by multiple daily additions of chemokines. These results should be of value in considering these chemokines for potential clinical use and for assessment of their mechanisms of action, alone and in combination. PMID- 7492627 TI - Hodgkin's disease developing after spontaneous remission of chronic lymphocytic leukemia. AB - We present a 71-year-old patient with chronic lymphocytic leukemia diagnosed 27 years ago. Initially, the disease was staged as Rai II and the patient suffered from secondary immunoglobulin deficiency. Nevertheless, no treatment was necessary at that time. Because of disease progression a single course of chemotherapy was given in 1984. During the following year there was a constant decline of the WBC, accompanied by normalization of the immunoglobulins; both have remained stable ever since that time. However, there was still residual bone marrow infiltration, indicating persisting CLL. In 1993 cervical lymphadenopathy occurred with acute onset. A diagnostic lymphadenectomy revealed Hodgkin's disease of the nodular-sclerosing subtype. The patient was staged as II-III according to the Ann Arbor Classification and underwent radiation therapy. Cytogenetic examination of the bone marrow revealed a normal karyotype with an inversion of chromosome 9. This case demonstrates the rate coincidence of two lymphoproliferative disorders in the same patient. The clinical course and the immunologic findings of this patient are presented, together with a review of the literature. PMID- 7492628 TI - Respiratory syncytial virus, a rare cause of severe pneumonia following bone marrow transplantation. AB - Mixed alveolar interstitial pneumonia is a much-feared complication in bone marrow transplant recipients because it carries a high mortality. Many cases in which an etiological agent is identified are due to fungi or cytomegalovirus; rarely, other infectious agents such as Pneumocystis carinii or (para)influenzavirus are involved. In this report we describe a patient who received intense chemotherapy followed by autologous bone marrow transplantation for relapse of testis seminoma and who developed a severe alveolar interstitial pneumonia caused by respiratory syncytial virus in the early posttransplant period. The patient fully recovered after treatment with ribavirin. PMID- 7492629 TI - Neutrophilic dermal infiltrates in granulocytopenic patients with acute leukemia. AB - Acute febrile neutrophilic dermatosis (AFND, Sweet's syndrome) is clinically characterized by fever, neutrophilic leukocytosis, and tender dermal plaques. Histological examination typically reveals infiltration of the dermis by neutrophils. In three patients (2 female, 1 male, 54-59 years) with acute leukemia (2 myelogenous, 1 lymphoblastic) dermal plaques developed during febrile episodes in chemotherapy-induced pancytopenia. The clinical appearance was compatible with AFND. The diagnosis was substantiated by skin biopsies which showed dense neutrophilic dermal infiltrates without leukemic cells. Leukocytoclastic vasculitis was considered as differential diagnosis. Plasma levels of soluble adhesion molecules ICAM-1, VCAM-1, and E-selectin regulating leukocyte transendothelial migration were in the normal range. Systemic glucocorticoids were avoided because of the high risk of infection during prolonged bone marrow aplasia. The lesions were treated with topical steroids and resolved without scarring within 1-5 weeks. AFND has been reported in association with acute leukemia at normal or elevated white blood cell counts. Although implausible from a pathophysiological point of view, similar neutrophilic dermal infiltrates were found in three patients during chemotherapy-induced pancytopenia with white blood cell counts distinctly below 1 x 10(9)/l. PMID- 7492631 TI - Organization of the sources of reticulospinal projections in the chicken by means of a confocal laser scanning microscope. AB - To investigate the manner of reticulospinal projections in chickens, two different fluorescent dyes (Fast-Blue and Diamidino-Yellow) were injected into the lumbar and sacral segments. Results showed that the main projections to the spinal segments are clearly different for the pons and medulla. The main source of the medulospinal projections was in the ventral reticular formation, and the projections from the dorsal reticular formation were sporadic. On the other hand, the main pontine projections arose from the dorsolateral tegmentum and they projected to the sacral segments. In contrast, projections to the lumbar segments were somewhat more dominant in the ventromedial tegmentum than in the dorsolateral tegmentum. Double projection neurons were observed in the ventrolateral reticular formation and dorsolateral tegmentum where there were many cells of origin of the spinal descending projections. The reticulospinal projections manner of birds was organized dorsoventrally, and it was comparable with that of mammals. PMID- 7492630 TI - Successful treatment of acute promyelocytic leukemia in pregnancy with all-trans retinoic acid. PMID- 7492632 TI - The effect of indomethacin on liver blood flow and oxygen supply-uptake relationship in the dog. AB - The effects of indomethacin on liver blood flow and oxygen supply-uptake relationship were investigated using a right heart bypass technique. Portal venous blood flow was decreased by the mesenteric vascular effects of indomethacin, which produce intense mesenteric vasoconstriction. Hepatic arterial blood flow was increased and therefore, total liver blood flow was not significantly changed after indomethacin administration. Portal venous oxygen delivery was significantly decreased by reductions in both portal venous blood flow and portal venous oxygen content. Total liver oxygen delivery, however, was not changed after indomethacin administration. This response was caused by a large increase in hepatic arterial oxygen delivery. Liver oxygen uptake and liver oxygen extraction ratio were not changed after indomethacin administration. We conclude, therefore, that total liver blood flow and oxygen delivery were well maintained, even if the mesenteric vascular effects of indomethacin decreased both portal venous blood flow and portal venous oxygen delivery. PMID- 7492633 TI - Erythroleukemia in two cats naturally infected with feline leukemia virus in the same household. AB - Erythroleukemia was observed in two unrelated cats infected with feline leukemia virus (FeLV) from the same household. Case 1, a 1-year-old neutered male cat developed erythroleukemia (M6) after a diagnosis of myelodysplastic syndrome (MDS Er) on the criteria of FAB classification of acute leukemias. Case 2, a 1-year old neutered female cat, which had close contact with Case 1, also developed erythroleukemia (M6Er). In both cases, marked proliferation of erythroid progenitor cells with disproportionally large numbers of immature forms was observed in the bone marrow. In Case 1, neoplastic proliferation of myeloid cells in the bone marrow was also noted at the terminal stage. Combination chemotherapy with daunomycin was partially effective for treatment of these erythroid neoplasias, but did not induce complete remission. Southern blot analysis using exogenous FeLV-specific probes indicated the clonal origin of these hematopoietic tumor cells. Furthermore, the erythroid and myeloid tumor cells in Case 1 were shown to be derived from independent transformed clones. A variant FeLV was shown to be integrated into the tumor cells in Case 1, while a full-length FeLV was found in both cases. Because these erythroid neoplastic diseases occurred in two unrelated cats kept in the same household and these diseases are rare, they may both have been associated with the same FeLV strain. PMID- 7492634 TI - Gas chromatography-mass spectrometric studies of canine urinary metabolism. AB - After the urine was treated with urease, lyophilized, and trimethylsilylated, it was examined for metabolic profiles in Dalmatian dogs and Shetland sheepdogs by gas chromatography-mass spectrometry (GC/MS), which simultaneously analyzes organic acids, amino acids, sugars, sugar alcohols, purine and pyrimidine bases, and nucleosides. The profiles were compared with those from human specimens. As clarified in past studies, Dalmatian dogs showed an extreme decrease in allantoin, which is the final product of purine metabolism in the canine of other species, and a marked detection of uric acid peak. This finding suggests that purine metabolism in Dalmatian dogs is different from that in the other species. Only two Shetland sheepdogs, whose mother had chronic renal failure, showed a marked excretion of uric acid, as in Dalmatian dogs. In addition, some Dalmatian dogs, who were maintained on a protein-restricted diet, showed a little excretion of uric acid. A large amount of uric acid is detected in combination with pentose monosaccharides, hexose-monosaccharides and sugar alcohols in neonatal human urine in comparison with the present dog samples. A marked difference between the canine and the humans is that phenylacetylglycine, which is derived from the aromatic amino acid phenylalanine, is excreted in the canine urine. Phenylacetylglycine is not detected in the human urine, and there have been no reports of its excretion in canine urine. PMID- 7492635 TI - Transmigration of fluid rapidly infused into dogs with renal blood vessels ligation and increased pulmonary capillary permeability. AB - To study the transmigration of fluid rapidly infused intravenously, lactated Ringer's solution was infused at the rate of 90 ml/kg/hr for 30 min into generally anesthetized dogs. Four groups were made; group C with no fluid infusion as the control, group F with rapid fluid infusion only, group LF with renal blood vessels ligation and rapid fluid infusion, and group OF with increased pulmonary capillary permeability experimentally induced by oleic acid and rapid fluid infusion. The extravascular lung water volume (ELWV) was measured by the thermal-sodium double indicator dilution method, and the amounts of ascites and urine, and other clinical parameters were monitored before and after fluid infusion. The results show that in the dogs without increased pulmonary capillary permeability excessive fluid is mainly excreted as urine (group F) or transudates into the abdominal cavity (group LF). But in the dogs with increased pulmonary capillary permeability (group OF) excessive fluid easily and rapidly transudated into the pulmonary interstitium, resulting in pulmonary edema. PMID- 7492636 TI - Stimulation of haptoglobin synthesis by interleukin-6 and tumor necrosis factor, but not by interleukin-1, in bovine primary cultured hepatocytes. AB - The hepatic synthesis of acute phase proteins in ruminants has been suggested to be regulated by some mechanisms different from those in other species such as rodents and human. To explore possible regulatory factors unique to ruminants, we examined effects of interleukin (IL)-6, IL-1 and tumor necrosis factor (TNF), on haptoglobin (Hp) synthesis using a primary culture system of bovine hepatocytes. After bovine primary cultured hepatocytes were incubated in the presence of various concentrations of the cytokines, the synthesis and mRNA level of haptoglobin and albumin were measured by labeling with [35S]-methionine and immunoprecipitation, and by Northern blot analysis, respectively. Hp synthesis was dose-dependently increased by recombinant human (rh) IL-6, and also by rhTNF alpha, but to a less extent, while it was not affected by rhIL-1 beta. The stimulatory effect is mainly pretranslational, because mRNA level of Hp changed in parallel with protein synthesis. In contrast, albumin synthesis was suppressed by these three cytokines similarly. These results are inconsistent with the previously proposed view that TNF and IL-1 overlap in their pathways leading to the transcriptional activation of many acute phase protein genes. In conclusion, there is a species-specific unique signaling system, especially for TNF, in transcriptional activation of bovine Hp gene. PMID- 7492637 TI - Relationship among seminal characteristics, fertility and suitability for semen preservation in draft stallions. AB - Seminal characteristics, fertility and the response to semen preservation (liquid storage and cryopreservation) were evaluated in 4 Draft stallions (Percheron 2, Breton 2). Seminal characteristics (gel-free volume, sperm concentration, sperm morphology, percentage of motile spermatozoa) were assessed in 5 ejaculates from each of the 4 stallions. The fertility of the stallions was calculated retrospectively as the accumulated pregnancy rate over 3 breeding seasons. Five ejaculates from each of the stallions were subjected to liquid storage at 5 degrees C. The percentage of motile spermatozoa (PMS) was assessed at 0, 24, 48 and 72 hr. Cryopreservation was done with a glucose-EDTA-lactose egg yolk extender and a skim milk-based extender containing clarified egg yolk (4 replicates per stallion). PMS and the percentage of spermatozoa with an intact plasma membrane (IPM) were evaluated before and after freezing. Although semen characteristics and fertility were within a physiological range, significant differences among stallions were found for gel-free volume, percentage of morphologically normal spermatozoa, PMS, and fertility but there was no clear relationship between these parameters. The response to liquid storage of diluted semen samples differed among stallions and best results were obtained in a stallion with the highest initial motility. The freezability of semen from the 4 stallions was poor with both of the freezing protocols (PMS range 13.8%-26.3%; IPM range 19.5%-38.0%). Results of liquid storage and cryopreservation were not correlated in individual stallions and were not related to their fertility. Since predictability for the response to semen preservation is low, the suitability of individual stallions for semen preservation has to be examined carefully.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7492638 TI - Characterization of chicken kit tyrosine kinase receptor in Cos cell transfectants and in chicken brain. AB - The Kit tyrosine kinase (Kit) encoded by the c-kit proto-oncogene is a receptor for stem cell factor (SCF). Kit proteins of mice and humans are expressed in various kinds of hematopoietic progenitor cells and are essential for the growth of these cells. Wild-type chKit (chKit+) and a mutant chKit (chKit42) that contained an amino acid change from Asp777 to Asn corresponding to that in Kit of the W42 mutant mice were produced in Cos-1 cells transfected with expression plasmids containing the chicken c-kit cDNA, and characterized using two kinds of anti-chKit antisera. The W42 mutant Kit has previously been shown to be defective for kinase activity. The chKit+ of 145 kilodalton (kDa) and 130 kDa with varying degrees of N-linked glycosylation were detected. Western blot analysis using an anti-phosphotyrosine monoclonal antibody showed that autophosphorylation of chKit+ was greatly enhanced upon chicken SCF induction. The chKit+ did not respond to mouse SCF. The kinase activity of chKit42 was abolished by the amino acid substitution, indicating the Asp777 residue was essential for the activity. In addition, 145 kDa chKit conjugated with sialic acid residue(s) was detected in chicken brain by immunoprecipitation using the antisera. An in vitro kinase assay showed the kinase activity of this protein. These structural and functional similarities of chKit to mammalian Kit proteins shown in this study implicate a possible role of chKit in chicken hematopoietic system. PMID- 7492639 TI - Brain lesions in chickens experimentally infected with a neuroadapted strain of mesogenic Newcastle disease virus. AB - Neuroadapted Newcastle disease virus (Q10) was selected by tenth serial passage, in the chicken brain of a mesogenic strain (Q0) originally isolated from quails. Specific pathogen-free birds were inoculated intranasally with one of these viruses. At daily intervals for 7 days and then at 10, 14, and 21 days post inoculation (PI), two birds from each group were killed and samples of the brain were collected for histopathological and virological examination. Q10 caused severe nonsuppurative encephalitis with nervous signs and high mortality. Lesions characterized by neuronal degeneration and necrosis, perivascular lymphocytic infiltration, and focal or diffuse astrogliosis occurred mainly in the parahippocampal cortex, hippocampus, hyperstriatum, neostriatum, subleptomeningeal and periventricular regions of the cerebrum. Spongy changes with neuronal degeneration and axonal spheroids were also observed in the brain stem of a few cases. The amount of virus in the brain reached a peak on day 4 PI and virus could not be recovered from the brain after 6 days PI. In contrast, Q0 caused nonfatal asymptomatic disease and virus could not be isolated from the brain, sections of which showed only minimal inflammatory changes. This difference in the lesions of the brain might be related to neurovirulence and, neuroadaptation by serial passage may occur by increased efficiency of viral replication in neurons. PMID- 7492641 TI - Morphometrical changes of the arterial walls of main arteries from heart to the abdomino-inguinal mammary glands of rat from virgin through pregnancy, lactation and post-weaning. AB - Morphometrical changes of the arterial walls from the ascending aorta to the abdomino-inguinal mammary glands of rat from virgin through pregnancy, lactation and post-weaning stages were investigated by using a light microscope and an image analyzer. Based on the histological architecture and organization of connective tissue fibers and smooth muscle cells in the media, the arterial segments were clearly identified into three types; elastic, transitional and muscular types. The ascending aorta, the aortic arch, the thoracic aorta and the abdominal aorta were identified as elastic type and the maximum number of elastic lamellae was recorded in the ascending aorta as 10-13. In elastic type of arteries, the tunica media was thicker than other tunics with the exception of the abdominal aorta, in which the tunica externa was a little bit thicker than that of the media. From the external iliac artery to the proximal part of the femoral artery, the arterial segments were transitional in type and the elastic lamellae were ranged from 1-3 in numbers. The deep circumflex iliac, the external pudendal, the caudal superficial epigastric, and the mammary arteries were identified as muscular type, and the media was completely devoid of elastic lamellae. Distinguishable significant changes of diameters and thickness of the arteries were recorded in the deep circumflex iliac, the external pudendal, the caudal superficial epigastric and the mammary arteries, and showed that the diameters and thickness were gradually increased during pregnancy and reached to the maximum values during lactation and again decreased following the post weaning stages.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7492640 TI - Flow cytometric analysis and immunohistochemistry of delayed-type hypersensitivity responses in mice immunized with rat hepatocytes. AB - Delayed-type hypersensitivity (DTH) responses to rat hepatocytes (HCs) in mice were investigated by flow cytometric analysis and immunohistochemistry with monoclonal antibodies directed against murine class II, CD4, and CD8 antigens. Mice were immunized subcutaneously (s.c.) with 10(6) rat HCs (referred to as s.c. immunized mice), and control mice were injected s.c. with sterile Hanks' solution (non-immunized mice). Four days later, 10(5) rat HCs were injected into the footpad of s.c.-immunized mice and non-immunized mice. The DTH response in s.c. immunized mice significantly increased after challenge when compared to that in non-immunized mice. The numbers of class II+, CD4+, and CD8+ cells in the footpad, and CD4+ and CD8+ cells in the inguinal lymph node of s.c.-immunized mice significantly increased during the DTH response. An increase in the number of CD4+ cells in the footpad of s.c.-immunized mice after challenge was more significant than that of non-immunized mice. The number of CD4+ cells increased more markedly in the footpad of s.c.-immunized mice as compared to that of CD8+ cells. Furthermore, immunohistochemical studies of the footpad of s.c.-immunized mice showed more severe infiltration of CD4+ cells rather than CD8+ cells at the injection site of rat HCs. These results suggest that the DTH response in the footpad of mice immunized with rat HCs is associated with severe infiltration of CD4+ cells. PMID- 7492642 TI - Relationship between degree of dehydration and serum electrolytes and acid-base status in cows with various abomasal disorders. AB - In 368 cows with various abomasal disorders--left abomasal displacement (LAD), right abomasal displacement (RAD), abomasal volvulus (AV) and abomasal impaction (vagal indigestion)--the degree of dehydration was compared with electrolytes and acid-base status of the cows. No clinical symptom of dehydration was observed in 62% of cases with LAD and 43% of cases with RAD. The cows were moderately and severely dehydrated in 86% of the cases with AV and 84% with abomasal impaction. Hypochloremic alkalosis was observed in cows with LAD only when the cows were severely dehydrated, but cows with RAD revealed less electrolytes and acid-base abnormalities despite the increase in the dehydration score. In cows with AV, the acid-base status could not be predicted by the degree of dehydration. Profound hypokalemia and hypochloremia were observed in cows with abomasal impaction for all dehydration states. These findings indicated that hypochloremic alkalosis could be predicted in cows with abomasal impaction for all dehydration states and in LAD cases with marked dehydration, but not in RAD cases. PMID- 7492643 TI - Efficacy of florfenicol on experimental Actinobacillus pleuropneumonia in pigs. AB - The efficacy of florfenicol was evaluated on experimental Actinobacillus pleuropneumonia in 5 to 7-week-old pigs. All pigs were intranasally inoculated with bacterial suspension containing 10(8) or 10(9) colony forming units of Actinobacillus pleuropneumoniae, Fukushima (serotype 2), Shope4074 (serotype 1), K17 (serotype 5), 8541 or 8543 (serotype 2; thiamphenicol-resistant strain). Florfenicol and thiamphenicol were given in feed for 12 days, from 5 days before inoculation to 7 days post inoculation (at necropsy). More than half the pigs died in the infected control groups, while no pigs died in florfenicol 50 ppm groups inoculated with each strain. On inoculation with thiamphenicol-sensitive strains, Fukushima, Shope4074 and K17, the average total score of clinical signs and percentage of lung lesion area in florfenicol 50 ppm groups were significantly lower than those in the infected control groups (P < 0.05). On inoculation with thiamphenicol-resistant strains, 8541 and 8543, the average total score of clinical signs and percentage of lung lesion area in florfenicol 50 ppm groups were significantly lower than those in the infected control and thiamphenicol 200 ppm groups (P < 0.05). The development of pleuropneumonia was remarkably prevented by medication with feed containing florfenicol in pigs inoculated with serotype 1, 2, 5 strains and thiamphenicol-resistant strains of A. pleuropneumoniae. PMID- 7492645 TI - Specular microscopy of corneal endothelial cells in rabbits. AB - The corneal endothelia of 19 male and 19 female Japanese White rabbits (age: 6 to 15 months) were examined with a contact-type specular microscope under general anesthesia. The specular microscopy showed uniform-sized, pentagonal or hexagonal endothelia arranged regularly on the innermost layer of the cornea. In addition, large endothelial cells were sporadically observed in some rabbits. These large cells had smooth contour, and were 3 to 15 times as large as the cells of standard size. The large cells were isolated from each other and randomly located in one or both eyes of both sexes in all age groups examined. The presence of large endothelial cells was not related with the wounds on the corneal surface or the inflammation in the anterior chamber. Concerning the cell density (cells/mm2) and the incidence of the large cell, no statistically significant difference was observed between the left and right eyes or between sexes. However, the cell density decreased in rabbits over 12 months old. PMID- 7492644 TI - Application of DNA fingerprinting with digoxigenated oligonucleotide probe (CAC)5 to analysis of the genetic variation within Taenia taeniaeformis. AB - DNA from T. taeniaeformis digested with the restriction endonuclease was hybridized with digoxigenated oligonucleotide probe (CAC)5. Metacestode and adult showed same clear multibanding patterns, which were characteristic of multilocus DNA fingerprinting. The fingerprinting patterns were quite different from those of the rodent hosts. Genetic variations in 4 laboratory-reared isolates of T. taeniaeformis, including 3 isolates which have been reported to be indistinguishable by infectivity, morphology and protein composition of metacestode, were investigated using this technique. Each of the 4 isolates exhibited isolate-specific fingerprinting patterns and were easily distinguished from one another, thus it was considered that (CAC)5 was a highly resolvable and informative probe for cestodes. However, it was also indicated that (CAC)5 was so sensitive that applying fingerprinting with (CAC)5 to taxonomical or phylogenetic analysis was limited where habitat of the host was restricted to the small area. In comparison to fingerprinting with 32P-labeled (CAC)5, fingerprinting with digoxigenated (CAC)5 represented more and sharper bands. It was considered that a digoxigenated probe was more useful for genetic analysis of cestodes. PMID- 7492646 TI - A genetic analysis of mixed population in Theileria sergenti stocks and isolates using allele-specific polymerase chain reaction. AB - In order to differentiate parasite populations bearing two allelic forms of p33/32, an immunodominant piroplasm surface protein of Theileria sergenti, two sets of oligonucleotide primers were designed to amplify either of the two allele by the polymerase chain reaction (PCR). Each set of the primers differentially amplified either of Ikeda- or Chitose-type p33/32 genes. By using this allele specific PCR and restriction enzyme digestion of amplified products, parasite populations within field isolates collected from different geographical regions in Japan were analyzed. Both of the allelic forms were detected in 13 samples out of 20 isolates and stocks by PCR. Either of Ikeda or Chitose type of p33/32 allele was detected in the other 7 samples. These results indicated that the majority of T. sergenti-infected calves (11/15) in Japan harbored mixed parasite populations bearing at least two different alleles of p33/32. PMID- 7492648 TI - The effects of dopamine and dobutamine on liver oxygen supply-uptake relationship in the dog. AB - The effects of dopamine and dobutamine in various doses on liver oxygen supply uptake relationship were studied in 12 mongrel dogs. Dopamine 3 and 7 micrograms/kg/min infusion rates and dobutamine 5 micrograms/kg/min infusion rate did not produce any changes in total liver oxygen delivery. On the contrary, total liver oxygen delivery was increased at the 15 micrograms/kg/min dopamine in fusion rate and dobutamine 10 and 15 micrograms/kg/min infusion rates. The ratio of total liver oxygen delivery to the systemic oxygen delivery was increased at the 15 micrograms/kg/min dopamine infusion rate. Liver oxygen extraction ratio was decreased at the 15 micrograms/kg/min dopamine infusion rate and at the same rate of dobutamine. These decreases were due to the increases in oxygen delivery while both oxygen uptakes were avariant from control levels. The results of this study demonstrated that high dose of dopamine (15 micrograms/kg/min) and medium and high doses of dobutamine (10 and 15 micrograms/kg/min) should be useful to increase the liver oxygen delivery. However, these increases in liver oxygen delivery during dopamine and dobutamine infusion were not associated with an improvement in liver oxygen metabolism, since liver oxygen uptake was not changed. PMID- 7492647 TI - Histological observation of the endometrium in repeat breeder cows. AB - The endometrium plays an important role during attachment and implantation of embryos. Using histological techniques, we evaluated the morphological changes of the endometrium in repeat breeder cows. Endometrial biopsy specimens were obtained from 5 Holstein repeat breeder cows, 5 normally cyclic Holstein cows and 5 normally cyclic Holstein heifers on Days 1 and 8 after estrus. On Day 1, in the repeat breeder cows, the glandular secretions and supranuclear vacuolation were observed, but glandular mitoses were not observed, whereas these secretory characteristics were not observed in the normal cows. However, the appearance of stomal mitoses, stromal edema and pseudodecidual reaction was observed as in the normal cyclic cows. On Day 8, in the repeat breeder cows, the characteristics on the glandular indices were not different from those on Day 1, and were similar to those of the normal cows on Day 8. But, the characteristics on the stromal indices, the stromal mitoses and pseudodecidual reaction, were not observed, although these characteristics were observed in the normally cyclic cows. The presence of advanced morphology in the gland on Day 1 and the stroma on Day 8 indicates that these two events are controlled independently. These endometrial asynchrony might result in cows repeat breeding. PMID- 7492649 TI - The selective effects of dopamine and dobutamine on liver circulation in the dog. AB - The selective effects od dopamine and dobutamine in various doses on liver circulation were studied in 12 mongrel dogs. Dopamine increased portal flow but decreased hepatic arterial flow markedly as infusion rate of dopamine increased. Dopamine 3 micrograms/kg/min infusion rate produced vasodilation in mesenteric vascular bed and the portal flow ratio to cardiac output was significantly increased. Dobutamine increased both portal and hepatic arterial flows at the 5 and 10 micrograms/kg/min dobutamine infusion rates, and decreased hepatic arterial flow at the 15 micrograms/kg/min dobutamine infusion rate. Both dopamine and dobutamine increased total liver flows, however, total liver flow ratio to cardiac output was not increased. Pressure gradient of portal system was not changed during dopamine and dobutamine infusion, since both portal venous pressure and hepatic venous pressure were avariant from control values. These findings suggest that congestive hyperemia was not occurred in intrahepatic portal vascular system when portal flows were increased during dopamine and dobutamine infusion. The results of this study demonstrate that both dopamine and dobutamine did not produce selective increases in total liver blood flow. In addition, both agents should be safe to use to the normal liver patient; total liver blood flow did not decrease and intrahepatic congestive hyperemia was not occurred when portal flow was increased. PMID- 7492651 TI - Effect of administration of serum thymic factor (FTS) in calves and rabbits infected with bovine immunodeficiency-like virus. AB - The effect of serum thymic factor (FTS) administration in bovine immunodeficiency like virus (BIV)-infected calves and rabbits was examined. We previously found that some of the macrophage functions were depressed and humoral immune responses against foreign proteins were delayed in BIV-infected calves compared to uninfected calves. After FTS administration, however, no delay of antibody responses against foreign proteins was observed in BIV-infected calves. Though the chemiluminescence (CL) responses of macrophages in BIV-infected calves were significantly depressed (p < 0.05), FTS administration resulted in the recovery of the CL responses in the BIV-infected calves comparable to those in the control calves. Antibody responses against foreign proteins in BIV-infected rabbits were significantly depressed (p < 0.025) as compared with those in uninfected rabbits, though the depression became no significant after FTS administration. PMID- 7492650 TI - Proliferative response and cytokine production of bovine peripheral blood mononuclear cells induced by the superantigens staphylococcal enterotoxins and toxic shock syndrome toxin-1. AB - The potential of staphylococcal enterotoxin A (SEA), B (SEB), C(SEC) and toxic shock syndrome toxin-1 (TSST-1) to act as superantigens by inducing polyclonal T cell mitogenesis and cytokine production was tested on bovine peripheral blood mononuclear cells (PBMC). These four toxins were capable of inducing strong proliferative response of PBMC from calves over a broad dosage range (1 pg/ml to 1 microgram/ml) in vitro. The toxin-activated blast cells consisted of both CD4+ T-cells and CD8+ T-cells, but the T-cell proliferation depended upon the presence of monocytes. Treatment of monocytes with monoclonal antibody to major histocompatibility complex class II antigens substantially inhibited the toxin induced T-cell proliferative response, but paraformaldehyde-fixation did not abrogate the accessory function. SEA, SEB, SEC and TSST-1, all induced the in vitro release of interleukin-2, interferon gamma and tumor necrosis factor alpha in a dose dependent manner. The results indicate that SEA, SEB, SEC and TSST-1 are capable of acting as superantigens by stimulating bovine T-cells as shown in the human and murine systems. The possible implications of these toxins in the immunopathogenesis of bovine mastitis caused by the infection with Staphylococcus aureus are discussed. PMID- 7492652 TI - Comparative hematological study of two breeds of foals (Andalusian and Arab) subjected to exercise of progressive intensity. AB - Exercise-induced hematological alterations were studied in 20 four-year old foals, 11 Andalusian and 9 Arabian. They were subjected to a test exercise program consisting of 4 levels of gradually increasing intensity (15, 20, 25 and 30 km/hr) with a duration of 5 min each. Blood samples were taken during resting, after each exercise level and at 10 and 30 min of recovery from exercise. The following hematic parameters were analyzed: red blood count, hematocrit value, hemoglobin, volumetric indexes (MCV, MCH and MHCH) and total plasma proteins. The alterations as a result of the physical effort were studied as well as any possible differences between the two breeds. The Arabian foals showed higher total plasma protein concentrations when resting, after each exercise level and at 10 mins of recovery time. The other variables studied remained with similar values in the two breeds without any significant changes. PMID- 7492654 TI - Effect of food on liver circulation in conscious dog. AB - Hepatic hemodynamics and the liver oxygen supply-uptake relationship, in response to eating, were investigated in a chronically catheterized conscious dog method. Portal venous pressure was significantly increased after eating, however was within the normal range reported previously. Hepatic venous pressure correlated well with portal venous pressure throughout the experiment, therefore, the pressure gradient of the portal system was unchanged. Hepatic venous oxygen content, correlated well with liver oxygen extraction, was unchanged after eating. Therefore, it is possible to assume that liver oxygen supply-uptake relationship is well maintained during digestion of food. PMID- 7492653 TI - Luteotropic effect of pregnant mare serum gonadotropin in cattle. AB - Pregnant mare serum gonadotropin (PMSG) is a long acting luteotropin with follitropin activity. Cattle were used to examine whether a smaller dose of PMSG has substantial luteotropic effect without excessive follitropic effects. Eleven Japanese Black heifers were randomly assigned to two groups. Animals were administered 500 IU PMSG on the day of ovulation (Day 0) to promote the formation of corpus luteum (group A; N = 5) or Day 7 to stimulate the luteal function (group B; N = 6). Four of them were given injections of saline on Days 0 and 7 of the preceding estrous cycle for control. All animals were examined by palpation per rectum every other morning and bled every day for steroids analyses. The length of estrous cycle was shortened by the treatment in group A compared with the previous cycle, whereas it was extended in group B (P < 0.05). Progesterone secretion was not enhanced in group A, but it was significantly elevated and sustained on higher levels in group B (P < 0.05) as compared with the control. Although estradiol-17 beta concentrations were significantly increased in both PMSG-treated groups (P < 0.05), no excessive follicular development was observed. It is concluded that 500 IU PMSG administered on Day 7 enhances luteal function without excessive follicular development, whereas the administration on Day 0 has an adverse effect on luteal function. PMID- 7492655 TI - Comparison of agar-gel precipitin responses among strains of fowl adenovirus using antigens prepared from chorioallantoic membranes and chicken kidney cell cultures. AB - Agar-gel precipitin responses obtained for serologically different strains of fowl adenovirus (FAV) in tests using antigens prepared from FAV-infected chorioallantoic membranes (CAM antigen) and chicken kidney cell cultures (CKC antigen) were compared. Findings showed that both types of antigens exhibited less sensitivity to heterologous than to homologous antisera and that quantitative differences in sensitivity were present between serotypes. CAM antigens were more sensitive than CKC antigens to heterologous antisera. Polyvalent CAM antigens containing 2 or 3 antigens increased sensitivity in testing of field serum samples, resulting in a higher rate of detection. PMID- 7492656 TI - Stimulatory effects of insulin on the development of bovine embryos fertilized in vitro. AB - To obtain information about the effects of insulin on the development of bovine embryos, fertilized bovine embryos in vitro were cultured in a chemically defined, protein-free medium: modified synthetic oviduct fluid (mSOF) supplemented with 1 mg/ml polyvinyl alcohol (PVA). The percentage of embryos reaching the morula stage was not affected by the addition of 5 micrograms/ml insulin alone, but insulin showed a beneficial synergistic effect with amino acids (AAs) on the embryonic development to morula. Supplementation of insulin (1 100 micrograms/ml) with mSOF+AAs did not improve the percentage of embryos developing to the blastocyst stage. However, a significant increase in cell numbers of blastocysts was observed with the addition of 1 mM glucose and 1 or 10 micrograms/ml insulin from the morula stage. These results show that insulin has a beneficial effect on the preimplantation development of bovine embryos in the presence of AAs and/or glucose, and suggest that insulin improves embryonic development by stimulating AAs transport and/or glucose uptake. PMID- 7492659 TI - Neurotensin-immunoreactive ganglion cells in the pigeon retina. AB - Neurotensin immunocytochemistry was carried out in the pigeon retina after colchicine had been injected into a ventricle of the brain. Immunoreactivity was observed in the ganglion cells and amacrine cells. The ganglion cells were stained in small and large types. This result indicates the presence of neurotensin in the ganglion cells of the pigeon retina. PMID- 7492660 TI - Bronchial ramification and lobular division of the lung of the Japanese badger (Meles meles anakuma). AB - The right lung of the Japanese badger (Meles meles anakuma) consists of the cranial, middle, caudal and accessory lobes. The left lung consists of the middle and caudal lobes. The Japanese badger has four bronchiole systems, dorsal, lateral, ventral and medial, on each side. The cranial lobe is formed by the first bronchiole of the dorsal bronchiole system. The middle lobe is formed by the first bronchiole of the lateral bronchiole system. The accessory lobe is formed by the first bronchiole of the ventral bronchiole system. The remaining bronchioles of the four bronchiole systems constitute the caudal lobe. PMID- 7492657 TI - A case report of the management of demodicosis in the golden hamster. AB - A 1.6 year-old male golden hamster was presented with alopecia on the back. The animal's physical condition was slightly below normal, and moderate pruritus was observed. Examination of skin scraping revealed mites identified as Demodex sp. Initially, combined therapy using 1% of selenium sulfide shampoo and a topical application of 0.013% amitraz was carried out, although this was not completely successful. Repeated use of 0.017% coumaphos was effective in achieving a complete cure of the alopecia associated with demodicosis. No adverse side effects were recognized during the treatment of this case. PMID- 7492658 TI - A case of multiple osteochondroma in the rat. AB - The occurrence of spontaneous osteochondroma in rats is extremely rare and only a few cases have been reported. Spontaneous multiple osteochondroma was found in a male SD strain rat, at the age of 58 weeks from the control group in a toxicity study. Histological findings of both a walnut-sized mass involving the humerus and scapula of right forelimb and a soybean-sized mass adjacent to the costochondrial junction of the right posterior rib were similar in appearance. The outer layer of the tumors consisted of hyaline cartilage and the core of the tumors consisted of trabecular bone with abundant fatty bone marrow. The periosteum of the surface was continuous with that of the parent bone, and cortical bone and the medullary cavity of the parent bone communicated with those of the tumors. Because of showing progressive enlargement, multiple osteochondromas in the rat were considered to be neoplastic in nature. PMID- 7492661 TI - Immunological responses to polyvalent canine vaccines in dogs. AB - The immunological responses to commercially available polyvalent vaccines in dogs were examined. There was a tendency in decreased lymphocyte counts on day 7 in the puppy and adult dogs. There was a significant increase in the blastogenesis of lymphocytes on day 7 and 21 in puppies, whereas no significant changes were seen in the adult dogs. Delayed type hypersensitivity (DTH) responses to phytohemagglutinin (PHA) and canine parvo-virus (CPV) vaccine monitored 0, 3, 8 weeks after vaccination produced strong reactions, in particular those to CPV vaccine rose significantly after vaccination and maintained the higher responses for at least 2 months. Therefore, it is considered that vaccination is immunomodulative rather than immunosuppressive and that DTH responses to PHA and CPV vaccine are helpful to monitor non-specific and specific immune functions in vivo. PMID- 7492662 TI - Application of the Limulus amoebocyte lysate test as an indicator of microbial contamination in pork carcasses. AB - The Limulus amoebocyte lysate (LAL) test, a simple and rapid method, was applied for the evaluation of bacterial contamination of carcasses in a slaughterhouse. Twenty-five pork carcasses were examined for bacterial contamination, and we evaluated the correlation between the LAL test and the methods usually employed for detecting indicator organisms of total viable cells, coliforms or Staphylococcus aureus. The reciprocal of the highest dilution found to be positive was used as the LAL index. A high correlation was observed between the LAL index and the number of coliforms, but not between the index and the number of other organisms. The LAL test was accomplished within 2 hr after sampling, and thus may serve as a useful tool for the substitute indicator of coliforms among bacterial contamination in carcasses. PMID- 7492663 TI - Rumen ciliates of Tokara native goat in Kagoshima, Japan. AB - The composition of rumen ciliate protozoa was surveyed in 15 Tokara native goats, Capra hircus domesticus, inhibiting Kagoshima, Japan. As a result, of 11 species with 3 forms under 5 genera detected, Entodinium simplex, E. nanellum and E. rectangulatum were found in all the animals examined. The large ophryoscolecid ciliates detected were Polyplastron multivesiculatum and Metadinium affine alone, which have been regarded as the typical members of Type A composition proposed by Eadie. Since Tokara native goats have been bred isolated in a small islet for a long time, native goats that had been kept in south Japan might possess Type A fauna. Average ciliate density was 43.9 x 10(4) per 1 ml of rumen fluid. The number of species per host was 8.3 on average. Diversity index was 1.412 on average. PMID- 7492664 TI - Fine structure of podocytes in the bovine renal corpuscle. AB - Thick primary processes issued from the hemispherical or plate-like cell bodies of podocytes in bovine renal corpuscles. These processes branched into secondary and tertiary ones, but no fine pedicels were observed on the outer surface of the podocytes by scanning electron microscopy, because each primary process closely interdigitated with those of adjacent cells. Many pedicels, however, were observed under the processes by using the freeze-fractograph and a transmission electron microscope. PMID- 7492665 TI - In vitro antibacterial activity of florfenicol against Actinobacillus pleuropneumoniae. AB - A total of 90 strains of Actinobacillus pleuropneumoniae isolated from porcine pneumonic lungs in Japan during 1989 to 1993 were investigated for their in vitro susceptibility to florfenicol (a derivative of thiamphenicol) and other drugs. The minimal inhibitory concentrations of florfenicol against A. pleuropneumoniae ranged from 0.2 to 1.56 micrograms/ml with a peak at 0.39 microgram/ml. Florfenicol showed a high antibacterial activity against A. pleuropneumoniae regardless of the sensitivity of the strains to thiamphenicol. PMID- 7492667 TI - Intrahepatic portal-venous changes in dogs with dirofilariasis: scanning electron microscopy of resin casts of vasculature. AB - Intra hepatic portal-venous changes in dogs with dirofilariasis, in which characteristic cavernous transformation has been histologically observed, were studied by scanning electron microscopy (SEM) of resin casts of vasculature. Changes in the central and sublobular veins were the most significant. The venous channels were extremely dilated, thin-walled and showed cavernous structure. SEM observation showed multi-saccular formation with beads-like, helical and extremely dilated configuration. The conducting venules, i.e. terminal tributaries of the portal vein, showed a rapid taper just before the point of merging into sinusoids. Neither vascular proliferation nor collateral circulation around the portal channels could be detected in the present resin cast study. PMID- 7492666 TI - Responsiveness of equine basilar artery to transmural nerve stimulation differs from that of porcine and bovine basilar arteries in vitro. AB - Transmural nerve stimulation (TNS) induced relaxations in porcine and bovine basilar arteries which were abolished by tetrodotoxin (TTX) and by L-nitro arginine (LNAG). However, TNS induced contractions in equine basilar artery which were abolished by TTX and by guanethidine, but not by LNAG. These results suggest that the TNS-induced contractions of equine basilar arteries may be mediated by norepinephrine release. PMID- 7492668 TI - Suppression of lymphocyte blastogenesis in cows with puerperal metritis and mastitis. AB - Mitogenic responses of peripheral blood lymphocytes in naturally occurring clinical puerperal metritis and mastitis were investigated. Glucose consumption index (GCI) values for phytohaemagglutinin (PHA), concanavalin A (Con A) and pokeweed mitogen (PWM) in the puerperal metritic cows and mastitic cows were significantly lower than those in the healthy cows. Suppression of lymphocyte blastogenesis was correlated to an increased concentration of serum ammonia in the puerperal metritic cows, and of alpha 1-acid glycoprotein (alpha 1AG) in the mastitic cows. Lymphocyte blastogenesis in the mastitic cows was also correlated to the serum concentration of vitamin E. These findings indicate that the puerperal metritic and mastitic cows are associated with impaired lymphocyte blastogenesis. PMID- 7492669 TI - Polymeric N-acetyl-D-glucosamine (chitin) induces prostaglandin E2 in dogs. AB - The level of prostaglandin E2 (PGE2) in the exudate induced by subcutaneous implantation of a complex formed from non-woven fabric of polyester (NWF) and polymeric N-acetyl-D-glucosamine (chitin) (chitin/NWF) or by implantation of NWF in dogs was measured by radioimmunoassay. The amount of PGE2 in the exudate induced by chitin/NWF was about five times as high as that in the exudate induced by NWF (p < 0.05), while the level of PGE2 in the exudate was similar to that in serum. PMID- 7492670 TI - Differences of phenotype and gene frequency by C system in canine species. AB - The canine C blood group was determined by an agglutination reaction with a plant agglutinin extracted from the seeds of Clerodendron trichotomum (CTL). A positive agglutination reaction was classified as type C and a negative reaction as type c. In this study, the C phenotype and the gene frequency of C were examined in 377 dogs (224 purebred dogs of 29 breeds and 153 mongrel dogs) that were bred in Utsunomiya-city, Tochigi Prefecture. Phenotype C was less frequent than phenotype c in every purebred dog. A variation in C gene frequency was observed among the purebred dogs. The highest frequency of C allele was in the Yorkshire Terrier (0.202), followed by Beagle, Shiba (0.091), Maltese (0.085), Shi Tzu (0.051) and Shetland Sheepdog (0.036). The C blood group system as a genetic marker may provide useful information for bleed identification, genetic studies between breeds, and pedigree certification. PMID- 7492672 TI - Effect of TCV-309, a novel platelet activating factor antagonist, on hemodynamics in dogs with endotoxin-induced shock. AB - The therapeutic effects of TCV-309, a novel platelet activating factor antagonist, on hemodynamics in endotoxin-induced shock were evaluated. Ten Beagle dogs were used under general anesthesia and artificial ventilation. After intravenous injection of endotoxin (3 mg/kg), TCV-309 (1 mg/kg) was administered intravenously to the dogs. The results showed that the depression of mean aortic pressure, cardiac output, left ventricular stroke work index and urine volume which occurred in endotoxin shock was significantly improved by administration of TCV-309. These results suggested that TCV-309 was a useful therapeutic for the circulatory disturbance in endotoxin shock. PMID- 7492671 TI - The phenotypes and gene frequencies of genetic markers in the blood of Japanese crossbred cats. AB - The phenotypes and gene frequencies of genetic markers in 8 blood group systems were studied in Japanese crossbred cats. The gene frequency was 0.051 for Ca, 0.949 for ca, 0.484 for TFF, 0.5133 for TFs, 0.279 for GCF, 0.721 for GCs, 0.945 for PGDA, 0.055 for PGDB, 0.544 for ESD1 and 0.456 for ESD2. Genetic polymorphism was not detected in the PGM, ACP and GLO systems. PMID- 7492673 TI - Infertility in a transgenic rat due to impairment of cytoplasmic elimination and sperm release from the Sertoli cells. AB - In line TGR(mRen2)26 transgenic rats (TGR26) bearing a randomly inserted additional renin transgene, the males, but not the females, were found to be infertile. Tissue was obtained from TGR26 males and littermate controls after perfusion fixation, and the morphology of the testes and epididymides was examined. Testis size was normal as was gross morphology, but careful examination revealed that the release of many spermatozoa at stage IX of the spermatogenic cycle was impaired. In addition, the process of cytoplasmic elimination was abnormal, as cytoplasmic fragments of elongate spermatids were present in the epididymis. In TGR26 males, seminiferous tubule lumen size was significantly larger (p < 0.001) than in littermate controls, a difference that was most marked at stages IX-XIV--an effect that could be related to the retention of spermatozoa. In situ hybridization confirmed that expression of renin mRNA could be detected in testes of TGR26 rats but not in normal controls or in a fertile line (TGR27) of rats bearing the same transgene. Immunocytochemistry and in situ and Northern hybridization were used to elucidate the pattern of expression of genes that previous studies have implicated in the process of sperm maturation and/or release. Of the gene products examined (sulphated glycoproteins 1 and 2 [SGP-1, SGP-2], transition proteins 1 and 2 [TP-1, TP-2], urokinase, and cyclic protein 2 [CP-2]), none showed any major change in the pattern of expression compared with that in controls. We postulate that TGR26 transgenic male rats may be infertile because the expression of a gene (or genes) involved in the process of cytoplasmic elimination and/or sperm release has been disrupted by the presence of the transgene close to or within the gene(s). Future planned studies will involve determination of the insertion site(s) and ultrastructural analysis of the final phases of spermiogenesis. PMID- 7492674 TI - Effects of indirect selection for pituitary responsiveness to gonadotropin releasing hormone on the storage and release of luteinizing hormone and follicle stimulating hormone in prepubertal male lambs. AB - Divergent selection based on the LH response to a 5-micrograms dose of GnRH in 10 wk-old Finn Dorset ram lambs has created two lines of sheep that differ significantly in their response to both physiological and pharmacological dosages of GnRH. The aims of this study were to investigate whether there are between line differences in endogenous LH and FSH secretion in ram lambs, as in the adult ewes, and to determine whether these differences are related to differences in storage of gonadotropins within the pituitary glands of the two lines. To address the first aim, endogenous LH and FSH secretion was monitored in 20 ram lambs from each line for a 6-h period at 2, 6, and 10 wk of age. To investigate potential between-line differences in gonadotropin storage, the LH responses to two acute challenges with either 0.5, 5.0, or 50.0 micrograms of GnRH were examined in 10 wk-old ram lambs from each line. The results demonstrated that the between-line difference in the LH response to a 5-micrograms GnRH challenge in ram lambs is accompanied by significant between-line differences in the regulation of endogenous gonadotropin secretion. Specifically, lambs from the High line secreted LH pulses of significantly greater amplitude than did lambs from the low line at 2 and 10 wk of age.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7492676 TI - Effects of relaxin on the endometrial stroma. Studies in mice. AB - The current study was designed to elucidate the effects of relaxin, administered systemically to mice in combination with estrogen and medroxyprogesterone, on the endometrial stroma, with use of structural, ultrastructural, and immunocytochemical methods. Female, sexually mature virgin mice were placed into 4 groups and treated with estrogen, estrogen + progestin, estrogen + relaxin, and estrogen + progestin + relaxin in sequence. The results obtained show that relaxin causes a striking increase in the protein-synthesizing machinery of endometrial stromal cells, leading them to resemble closely the decidualized cells of rodent pregnant uterus. Relaxin also induces in the same cells the expression of laminin, which is regarded as a reliable marker of decidual transformation of endometrial stromal cells in mice, and which is needed for trophoblast to adhere to and invade the endometrial stroma. Moreover, relaxin seems to synergize with the steroids in promoting a growth response of the endometrial stromal cells. The above findings allow for relaxin to be regarded as an agent capable of promoting decidual transformation of endometrial stromal cells and, possibly, to participate in the events that lead to blastocyst implantation. PMID- 7492675 TI - Influence of a cigarette smoke extract on the hormonal regulation of platelet activating factor acetylhydrolase in rats. AB - We evaluated the effects of estrogen, progestin, and cigarette smoke extract (CSE) on plasma platelet-activating factor (PAF) acetylhydrolase (AH; PAF-AH) activity in rats. The effects on rat tissues of an i.v. injection of PAF were studied as part of our investigation of the mechanisms involved in thrombotic episodes. Plasma PAF-AH activity in adult female rats (14 wk of age) treated with 17 alpha-ethynylestradiol (100 micrograms/kg, 5 days) was decreased by 70%. Medroxyprogesterone (50 mg/kg, 5 days) increased PAF-AH activity by 50%. CSE (0.5 cigarette/kg, 5 days) did not alter PAF-AH activity during the treatment. However, a combination of CSE and 17 alpha-ethynylestradiol decreased plasma PAF AH activity by 90%. The decrease in PAF-AH activity was age-dependent. The effect of medroxyprogesterone on plasma PAF-AH activity was not influenced by CSE. When PAF (5-40 nmol/kg) was injected i.v. into untreated adult female rats, 9 of 16 animals died after a 20-nmol/kg dose of PAF. Macroscopic findings included hemorrhage, hyperemia, and congestion in the lungs and heart, and necrosis-like changes in the gastrointestinal tract. Microscopically, thrombi were observed in the lungs and heart. When PAF was administered to adult female rats pretreated with sex steroid hormones, the mortality of rats with low plasma PAF-AH activity caused by 17 alpha-ethynylestradiol was increased but that of rats with high PAF AH activity caused by medroxyprogesterone was decreased. Thus, PAF and PAF-AH may play important roles in the thrombotic episodes known to occur in female smokers who use oral contraceptives. PMID- 7492677 TI - Are immediate early genes involved in gonadotropin-releasing hormone receptor gene regulation? Characterization of changes in GnRH receptor (GnRH-R), c-fos, and c-jun messenger ribonucleic acids during the ovine estrous cycle. AB - GnRH regulates the secretion and synthesis of gonadotropins by binding to specific receptors located in the plasma membrane of the pituitary gonadotroph. Like the concentration of the signaling ligand GnRH, the number of GnRH receptors (GnRH-R) varies dynamically with the changing endocrine milieu during the ovine estrous cycle. With the recent success in cloning of the mammalian GnRH-R gene, it is becoming increasingly evident that some of the changes in GnRH-R numbers may be mediated at least in part via changes in GnRH-R gene transcription. However, the regulatory steps involved in the GnRH-R transcription are unknown. The present studies were conducted to 1) characterize in detail the changes in GnRH-R gene expression during the 16-day ovine estrous cycle, 2) determine whether or not changes in GnRH-R gene expression during the estrous cycle are paralleled by alterations in the expression of c-fos and c-jun mRNAs, and 3) determine whether GnRH can induce expression of c-fos and c-jun mRNAs. Results revealed that concentrations of GnRH-R mRNA were highest on the day before estrus, when circulating LH concentrations were still low. GnRH-R mRNA concentrations declined steadily starting at 5 h postestrus, the time of the preovulatory LH surge, reaching their lowest levels by 24 h after estrus. Changes in c-jun mRNA levels, in general, paralleled changes in GnRH-R mRNA concentrations, being highest on the day before estrus and declining thereafter. c-Fos mRNA followed a different time course than c-jun mRNA, remaining elevated from Day 8 prior to estrus until the onset of estrus.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7492678 TI - Parthenogenic activation of pig oocytes by protein kinase inhibition. AB - Previous studies have shown that protein kinase stimulators can induce the release of the metaphase II arrest in mouse ova. The present report is about the role of protein kinase in parthenogenic activation of pig oocytes, which was studied using a broad-spectrum protein kinase inhibitor. Metaphase II oocytes were obtained via in vitro maturation. Two sources of H7 [1-(5 isoquinolinesulfonyl)-2-methylpiperazine, HCl] were tested--Sigma (H7s) and Calbiochem (H7c). Both were found to be equally effective in promoting release of the metaphase II block. Activation, release of the metaphase II arrest, and progression to the first interphase could be induced at the highest percentage with an exposure to 2.0 mM H7s for 80 min (68.1%). In another experiment, H7c resulted in 69.5% activation, while iso-H7 [1-(5-isoquinolinesulfonyl)-3 methylpiperazine, HCl] at a similar concentration and exposure duration resulted in 25.7% activation. H7s and H7c were more effective than iso-H7 in inducing the appearance of a 22-kDa protein that is associated with normal fertilization in the pig. In contrast, although pronuclei could form and the protein profiles were indicative of activation, cortical granule exocytosis did not occur, and oocytes failed to develop to the blastocyst stage after H7 treatment. In contrast to the H7-treated oocytes, electrostimulation resulted in pronuclear formation, the appearance of the 22-kDa protein, release of cortical granules, and development to the blastocyst stage. These data demonstrate that broad-spectrum inhibitors of protein kinase are unable to induce all the events associated with normal fertilization. PMID- 7492679 TI - Viability assessment of mammalian sperm using SYBR-14 and propidium iodide. AB - The proportion of living sperm in semen from six representative mammals was assessed by means of a dual staining technique using the stains SYBR-14 and propidium iodide (PI). SYBR-14, a newly developed fluorescent nucleic acid stain, maximally absorbs at 488 nm and emits at 518 nm when bound to DNA. Microscopic examination revealed that SYBR-14 stained the nuclei of living sperm bright green as determined by simultaneous examination of fluorescence and motility. Conversely, PI stained only nonmotile sperm that had lost their membrane integrity. Sperm from bulls, boars, rams, rabbits, mice, and men were stained and examined through use of fluorescence microscopy. The proportions of living and dead sperm were determined by first staining with SYBR-14 and PI and then assessing stain uptake by flow cytometry. Similar staining patterns were observed in all six mammalian species tested. Three populations of sperm were identified: living--SYBR-14 stained, dead--PI stained, and moribund--doubly stained. The SYBR 14 staining was replaced by PI staining as sperm progressed from living to moribund. The transition from green (SYBR-14) to red (PI) fluorescence started at the posterior region of the sperm head and proceeded anteriorly. The proportions of living and dead sperm in mammalian semen were readily identified through use of dual staining with SYBR-14 and PI and quantified through use of flow cytometry. PMID- 7492680 TI - Molecular cloning and characterization of a mouse oviduct-specific glycoprotein. AB - In the present study, we have isolated the cDNA for the mouse oviduct-specific glycoprotein (MOGP) by screening the mouse oviduct cDNA library with the bovine oviduct-specific glycoprotein (BOGP)-cDNA probe and by the 5' rapid amplification of the cDNA end (5'RACE). The total length of cDNA was determined to be 2525 base pairs (bp) by sequence analysis. The coding region contained 2163 bp translating to 721 amino acids. Based on comparisons with the N-terminal amino acid sequences of purified-BOGP and of hamster oviduct-specific glycoprotein (oviductin), it was inferred that the derived amino acid sequence contained a signal peptide region of 21 amino acids and a mature MOGP (core protein) region of 700 amino acids (76,515 daltons). It was also inferred that the mature MOGP contained three potential N-linked glycosylation sites and 24 possible O-linked glycosylation sites, and had the unique seven-residue repeat sequence (21 repeats) within the predicted sequence in the C-terminal side. The amino acid sequence of a portion of MOGP was highly homologous to that of BOGP (71% identity), baboon oviduct specific glycoprotein (61% identity), and human oviduct-specific glycoprotein (77% identity). Significant homologies were also observed with two mammalian secretory proteins that were reported as a mammalian member of a chitinase protein family. Northern blot hybridization with a DIG-labeled probe indicated that a single message of 2.8 kb was present in total RNA prepared from oviductal tissue. In situ hybridization using MOGP-cDNA showed that a MOGP message was only detected in the oviductal epithelial cells. These results strongly suggest that a significant degree of homology exists among oviduct-specific glycoproteins of various mammalian species. PMID- 7492683 TI - Mast cell and eosinophil distribution and activation in human endometrium throughout the menstrual cycle. AB - Tryptase and chymase immunolocalization techniques have been used to examine the distribution, activation, and tryptase/chymase phenotype of mast cells (MCs) in 107 endometrial specimens that represented every day of the human menstrual cycle. MCs were identified in the endometrium in all stages of the menstrual cycle; similar MC numbers were observed for the functionalis, basalis, and muscularis. Extensive MC activation/degranulation, as judged by extracellular tryptase, was a common feature of the functionalis in specimens sampled just prior to and during menstruation. MC activation was also prominent in the functionalis at times coincident with recognized stromal edema. MCs of the functionalis did not contain chymase; all stained for tryptase and represent the MCT phenotype. By contrast, the basalis and muscularis showed a proportion of MCs containing both tryptase and chymase, MCTC. One important function for extracellular MC tryptase and chymase is their ability to activate precursor forms of the matrix metalloproteinases, enzymes recognized as instrumental in stromal degradation. Quantitative analysis of MC numbers, expressed relative to stromal cell numbers/mm2, indicated no major changes during the menstrual cycle, although changes in MC morphology, granule content, and activation/degranulation were recognized for specific stages. Eosinophils, detected with monoclonal antibodies EG1 and EG2, were absent from extravascular sites between Days 5 and 26 but showed local accumulations just prior to and during menstruation. Since MCs and eosinophils between them contain a variety of potent mediators, it seems likely that both cell types assume important functional roles in relation to tissue and vascular remodeling associated with endometrial physiology. PMID- 7492681 TI - Proliferating cell nuclear antigen marks the initiation of follicular growth in the rat. AB - Despite expanding knowledge on the kinetic aspects of folliculogenesis, the question of what initiates follicle growth remains unanswered. Efforts to solve this problem have been thwarted by the absence of sensitive markers to identify the onset of follicular growth. In this study, we determined whether increased proliferating cell nuclear antigen (PCNA) correlates with initiation of follicle growth and might therefore be useful for studying early events in this process. Paraffin sections of ovaries from cycling adult rats, prepubertal eCG+hCG-primed rats, and prepubertal control rats were processed for immunocytochemistry by use of a PCNA primary antibody. In primordial follicles, neither granulosa cells nor oocytes stained for PCNA. PCNA immunoreactivity coincided with the earliest sign of follicle growth, appearing in pregranulosa cells of early primary follicles just beginning to grow. In all primary follicles, some granulosa cells were PCNA positive. PCNA immunoreactivity in oocytes first appeared in primary follicles, preceding oocyte enlargement. In preantral and antral follicles, granulosa cell PCNA staining was uniform throughout the granulosa cell layers. Oocytes were positive for PCNA in both preantral and antral follicles. PCNA expression diminished in atretic follicles. In CL, granulosa cell PCNA expression was also decreased. Theca cell PCNA expression was first evident in the transitional follicle (1-2-layer granulosa cells) and was present in all stages thereafter. The pattern of PCNA expression did not differ among adult cycling, prepubertal eCG+hCG-stimulated, and control rat ovaries.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7492682 TI - Tissue inhibitors of metalloproteinases in endometrium of ovariectomized steroid treated ewes and during the estrous cycle and early pregnancy. AB - Tissue inhibitors of metalloproteinases (TIMPs) have an important role in remodeling of tissues and are likely to be implicated in uterine function, including embryo implantation and placentation. Expression of mRNA for TIMP-1 and TIMP-2 was examined by Northern analysis of endometrial RNA derived from steroid treated ovariectomized ewes and from intact ewes during the estrous cycle and early pregnancy. Expression of mRNA for TIMP-1 (transcript size 0.9 kb), high in ovariectomized ewes, was substantially reduced by estrogen and to a lesser extent by progesterone. In cyclic and pregnant animals, abundance remained low until Day 10 and then increased, with high abundance continuing to Day 20 in the pregnant animals. Two transcripts for TIMP-2 were detected in ovine tissues--the 3.5-kb transcript and, in greater abundance, the 1.0-kb transcript. In ovariectomized ewes, endometrial abundance of both transcripts was low, and it decreased following estrogen treatment but was stimulated by progesterone alone or progesterone in the presence of estrogen. Abundance of TIMP-2 mRNA increased from Day 4 to Day 14 of the cycle. During early pregnancy, expression of the 1.0-kb transcript increased from Day 4 to Days 12-14 and was maintained at a high level to Day 20, whereas the 3.5-kb transcript decreased after Day 14 to very low levels by Day 20. In contrast with this pattern of regulated expression of TIMP, mRNA for proMMP-1 and for proMMP-3 was not detectable in any of the same tissues by Northern analysis. TIMP-1 protein was immunolocalized to both epithelium and stroma of intact endometrium, and the intensity of immunostaining was correlated with mRNA levels. TIMP-1 was secreted by both epithelial and stromal cells in primary culture, and its identity was confirmed by Western analysis, while reverse zymography demonstrated TIMP-1 and TIMP-2 along with a putative ovine TIMP-3 in the culture medium from both cell types. The precise role of TIMP in the endometrium remains to be established. PMID- 7492684 TI - Striking changes in the structure and organization of rat fetal membranes precede parturition. AB - Premature rupture of fetal membranes can harm infant and mother. It is unclear whether structural changes predispose these membranes to breaking. We thus assessed rat visceral yolk sac placenta (VYSP) and amnion by light and by transmission electron microscopy on Days 18-21 of gestation. Light microscope sections were stained for connective tissue (extracellular matrix) components: collagen, glycoprotein, and glycosaminoglycans/proteoglycans. Some tissue was incubated with chondroitinase ABC. We observed that fetal membranes became increasingly fragile, rupturing readily on Day 21. On Days 18-20, the two epithelial layers of the capsular VYSP were separated by a well-developed, well vascularized connective tissue layer that stained intensely for all matrix components studied; on Day 21, the connective tissue layer was thinner, moderately stained, and less vascularized. On Days 18-20, the two cellular layers of the amnion were separated by a narrow, compact connective tissue layer that stained modestly for all matrix components; on Day 21, this area was widened and stained faintly. Transmission electron microscopy showed that collagen fibrils of the amnion were abundant, closely packed, and well organized on Days 18-20, whereas on Day 21 they were few in number, widely spaced, and disorganized. Similar changes were present after incubation with chondroitinase ABC. In addition, amniotic epithelial cells were moribund and delaminating, basal laminae were deteriorating or absent, and few cells were at the outer surface of the amnion. All changes preceded parturition. We conclude that the structural integrity of rat fetal membranes is impaired before birth through the loss of connective tissue components and cells, changes that presumably underlie membrane rupture. Lastly, the similarity of structural changes in rat and human fetal membranes point to the potential usefulness of the rat model. PMID- 7492685 TI - 92-kDa gelatinase (matrix metalloproteinase-9) is induced in rat amnion immediately prior to parturition. AB - The fetal membranes undergo striking changes in structure before delivery that involve catabolism of the extracellular matrix. To investigate the role of specific enzymes in this process, we examined gelatinase activities in rat amnion, visceral yolk sac placenta, and placenta and amniotic fluid between Days 18-21 of pregnancy. Matrix metalloproteinase (MMP)-2 was present in amnion on all days, and its activity increased slightly on Day 21. The 92-kDa gelatinase, MMP 9, was not detected on Days 18-20 but appeared by the morning of Day 21. There was a marked increase in MMP-9 mRNA in the amnion on Day 20, preceding the appearance of MMP-9 activity. Western blotting confirmed an increase in MMP-9 protein in amnion on Day 21. MMP-2 and MMP-9 activities were detected in extracts of whole yolk sac placenta, placenta, and amniotic fluid, but there were no striking changes in these gelatinases between Days 18 and 21. However, the capsular regions of the visceral yolk sac placentae, which thin and rupture during labor, did show higher MMP-9 activity on Day 21 than on Days 18 and 20. We suggest that the striking increase in MMP-9 expression in amnion and possibly the capsular region of the visceral yolk sac placenta approximately 12 h prior to delivery is responsible, in part, for the alterations in the structure of these fetal membranes before parturition. PMID- 7492686 TI - Molecular characterization of a hamster oviduct-specific glycoprotein. AB - There is growing evidence that the oviduct is not a passive conduit for gamete and embryo transport but serves a function for the gametes and/or embryos. The oviductal epithelium secretes one or more specific glycoproteins that associate with the egg after ovulation. Several published reports including our preliminary studies have suggested that the egg-associating glycoprotein(s) from the oviduct exists in several mammalian species including golden hamster. However, little or almost no biochemical characterization of the hamster oviduct-specific glycoprotein (HOGP) has been reported. To analyze the molecular structure of the HOGP in detail, we have attempted molecular cloning of cDNA corresponding to HOGP. A cDNA library constructed from the hamster oviduct in the phage vector lambda ZAPII was screened with digoxigenin-labeled, baboon oviduct-specific glycoprotein cDNA as the probe. A single positive clone was isolated, and the nucleotide sequence of the isolated cDNA was determined. Rapid amplification of cDNA end was carried out to obtain a proximal 5' cDNA end of the clone. The cDNA clone consisted of 2387 bp, and the coding region contained 2013 bp translating to 671 amino acids. The amino acid sequence deduced from the cDNA sequence confirmed the chemically determined NH2-terminal sequence of a HOGP and suggested that the derived amino acid sequence contained a signal peptide region (21 amino acids) and 650 amino acids (70,890 daltons) of the mature form of the HOGP region. The amino acid sequence of HOGP appeared to have eight potential N glycosylation sites. Northern blot analysis revealed that a single message of approximately 2.5 kb was present in oviductal RNA but not in the RNA of several other hamster tissues. The HOGP showed high amino acid sequence homology with baboon, bovine, and human oviduct-specific glycoprotein. These results demonstrate that an oviduct-specific glycoprotein homologue gene exists in various mammalian species including rodent. PMID- 7492687 TI - Progestin and antiprogestin differentially regulate the expression of insulin like growth factors (IGF-I and IGF-II) messenger ribonucleic acid in human endometrial stromal cells. AB - Previous studies have shown that insulin-like growth factors (IGF-I and IGF-II) stimulate mitogenic activity in human endometrial stromal cells. In the present study, we have investigated the expression of IGF-I and -II mRNA to ascertain any autocrine growth-promoting effect in this system. Northern blot analysis revealed that endometrial stromal cells express multiple sizes of IGF-I and -II transcripts. The effect of progestin and antiprogestin was studied during decidualization of endometrial stromal cells in long-term culture. Solution hybridization and a ribonuclease protection assay of control cells revealed that the level of IGF-I mRNA was low, whereas IGF-II mRNA was always abundant. Medroxyprogesterone acetate (MPA) stimulated the expression of IGF-I mRNA > 4 fold in predecidualized cells during the first 10 days of culture. IGF-I mRNA decreased to basal level in prolonged culture when cells were decidualized. In contrast, MPA suppressed the IGF-II mRNA level by 60% in predecidualized cells, but IGF-II mRNA was highly expressed after 20 days of incubation with MPA (5-fold increase from Days 5-10 to Day 20 of culture). In progestin-pretreated cells, addition of the antiprogestin RU486 for 1-4 days reduced IGF-I mRNA by 50-90%. RU486 reversed the suppressive effect of MPA and increased IGF-II mRNA. This study indicates that progestin and antiprogestin differentially regulate IGF-I and IGF-II mRNA levels in human endometrial stromal cells. PMID- 7492688 TI - Gonadal responses of the male tau mutant Syrian hamster to short-day-like programmed infusions of melatonin. AB - Changes in the duration of the nocturnal peak of the circadian rhythm of melatonin secretion by the pineal gland mediate pohotoperiodic control of reproductive cycles in mammals. The present study examined whether the tau mutation of the circadian clock of the Syrian hamster altered the ability of the animals to exhibit photoperiodic, gonadal responses to timed infusions of exogenous melatonin. Pinealectomized tau mutant and wild-type hamsters received s.c. infusions of vehicle or melatonin solution (50 ng/h) for either 8 h or 6.67 h (i.e., 83% of 8 h, a reduction by the same proportion as circadian period is shortened by the mutation, 20 h vs. 24 h) once every 20 h for 6 wk. Testicular weight and serum levels of LH and prolactin were measured to assess the effects of the infusions on the reproductive axis. Both genotypes exhibited significant gonadal regression to the 8 h and 6.67 h melatonin signals, versus saline controls, although in taus the magnitude of the response to 6.67 h melatonin was significantly greater and more consistent than in wild-types. Compared to saline infused hamsters, animals that received infusions of melatonin of either duration showed significantly reduced serum levels of LH and prolactin. There was no significant difference between the two genotypes in the observed endocrine responses to melatonin infusions. These preliminary data demonstrate that tau mutant hamsters can respond to programmed systemic infusions of melatonin in a manner comparable to that observed in wild-type hamsters, and therefore, that the tau mutation of the circadian clock has not impaired the ability of the hamster to measure photoperiodic time, as represented by melatonin. PMID- 7492689 TI - Mouse growth hormone-releasing factor secretion is activated by inhibin and inhibited by activin in placenta. AB - We investigated the effects of activin and inhibin on the regulation of mouse growth hormone-releasing factor (mGHRF) secretion by primary placental cells harvested at Day 12 of pregnancy. Activin-A, an activator of FSH secretion, inhibited mGHRF secretion. In contrast, inhibin, an inhibitor of FSH secretion, activated mGHRF secretion. The lowest concentrations of activin-A and inhibin that significantly affected mGHRF secretion were 2 nM. Follistatin, a binding protein of activin, completely eliminated the ability of activin to inhibit mGHRF secretion. The steady-state level of mGHRF mRNA, as assessed by Northern analysis, was reduced by incubation of placental cells with activin-A. All activin and inhibin subunit mRNAs were expressed in mouse placenta, and their expressions increased during gestation. These findings suggest that activin and inhibin have opposite effects on mGHRF secretion as compared with FSH secretion and that they regulate mGHRF secretion in an autocrine or paracrine manner in the mouse placenta in vivo. PMID- 7492690 TI - Posttranscriptional inhibition of mouse placental lactogen-II secretion by transforming growth factor beta 1: synergistic effects with epidermal growth factor and interleukin-6. AB - We studied the effect of transforming growth factor beta 1 (TGF beta 1) on mouse placental lactogen (mPL)-I and mPL-II secretion by primary cultures of placental cells from Days 7, 9, and 12 of pregnancy. We also studied the effects of co incubation of epidermal growth factor (EGF) or interleukin-6 (IL-6) with TGF beta 1 on mPL-I and mPL-II secretion. TGF beta 1 at 10 ng/ml did not affect mPL-I secretion by cells from Days 7 or 9 of pregnancy or mPL-II secretion by cells from Day 7 of pregnancy but significantly inhibited mPL-II secretion by cells from Days 9 or 12 of pregnancy. The lowest concentration of TGF beta 1 that significantly inhibited mPL-II secretion by cells from Days 9 or 12 of pregnancy was 1 ng/ml. Immunocytochemistry for mPL-II indicated that treatment of placental cells from Day 12 of pregnancy with 10 ng/ml TGF beta 1 significantly reduced the number of mPL-II-containing cells. Inhibition of mPL-II secretion by TGF beta 1 was eliminated completely by addition of an anti-TGF beta 1 antibody. Northern analysis showed that steady state levels of mPL-II mRNA were not reduced by incubation of placental cells from Day 12 of pregnancy with 10 ng/ml TGF beta 1 for 5 days. EGF at 10 ng/ml significantly inhibited mPL-II secretion by cells from Day 7 of pregnancy, and addition of 10 ng/ml TGF beta 1, which did not itself inhibit mPL-II secretion by those cells, enhanced the inhibition by EGF of mPL-II secretion.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7492692 TI - Inhibition of deoxyribonucleic acid synthesis during premeiotic stages of spermatogenesis by a factor from testis-associated lymphomyeloid tissue in the dogfish shark (Squalus acanthias). AB - To investigate factors controlling spermatogonial proliferation, we used premeiotic (PrM) spermatocysts (stage-synchronized germ cell/Sertoli cell clones) derived from the testis of the dogfish shark (Squalus acanthias) as an in vitro test system to estimate DNA synthesis by [3H]thymidine incorporation. Coculture of PrM spermatocysts with spermatocysts of the same (PrM) or more advanced stages (M, meiotic; PoM, postmeiotic) revealed the presence of an increasing gradient of inhibitory bioactivity from immature to mature stages (PoM > M > PrM). An even more potent and effective inhibition was detected when PrM spermatocysts were cocultured with equivalent amounts of epigonal organ, a lymphomyeloid tissue encapsulating the testis adjacent to the mature (PoM) region and immediately upstream in the vascular pathway. Inhibitory bioactivity also was present in spent media from cultured epigonal fragments and in cytosolic subfractions of epigonal homogenates but was undetectable in epididymis, muscle, serum, and red blood cells. Lower but significant inhibition was obtained with the white blood cell fraction of peripheral blood in one of two experiments. Effects of the epigonal growth-inhibitory factor (EGIF) were dose- and time-dependent, had a short response latency (3 h), were completely reversible (< 24 h after washout), and counteracted but did not block the stimulatory effects of insulin on [3H]thymidine incorporation by PrM spermatocysts. EGIF was equally inhibitory when tested on each of five PrM substages (stem cells-->preleptotene). Analysis of the entire series of experiments showed that testis-derived inhibitory activity varied seasonally, with maximum effectiveness correlated with periods of spermatogenic inactivity.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7492691 TI - Luteinizing hormone/chorionic gonadotropin bioactivity in the common marmoset (Callithrix jacchus) is due to a chorionic gonadotropin molecule with a structure intermediate between human chorionic gonadotropin and human luteinizing hormone. AB - Chorionic gonadotropin (CG), a pregnancy-specific heterodimeric hormone found in primates, is responsible for CL rescue with pregnancy maintenance. Of the primates, the human and baboon gene sequences are the only structures so far determined. In order to study the structure and function of CG in other primates, we have isolated and sequenced the coding regions for the two subunits of marmoset CG (mCG) by the reverse transcription/polymerase chain reaction method. Study of multiple clones confirmed a high degree of homology with the human sequences (88% and 80% for the alpha and beta nucleotide sequences, respectively). Marmoset CG alpha has an extra four amino acids compared to hCG alpha, whereas the mCG beta sequence has a 3-bp deletion that maintains the reading frame and C-terminal amino acid sequence. Most of the differences between hCG beta and mCG beta peptides occur in the C-terminal region, which includes the loss of two of the O-linked glycosylation consensus sequences and the presence of an N-linked glycosylation consensus sequence. When mCG alpha and beta were co expressed in CHO cells, assembly of biologically active hormone was confirmed by induced steroid secretion by MA10 cells. Partially purified mCG beta was used to raise anti-mCG antibodies. To date, an antibody has been obtained that is capable of detecting recombinant mCG beta, recombinant mCG dimer, and mCG dimer secreted by cultured marmoset trophoblast. Marmoset CG alpha and beta were also detectable at the transcriptional level in cultured trophoblast by in situ hybridization. This suggests that the LH/CG bioactivity reported from marmoset placentae and embryos is due to a molecule with structural features common to hLH (glycosylation pattern) and hCG (CG beta C-terminal structure). PMID- 7492693 TI - Cytokines that use gp130 as a signal transducer stimulate mouse placental lactogen-I (mPL-I) but inhibit mPL-II production in vitro. AB - Interleukin-11 (IL-11), leukemia inhibitory factor (LIF), and oncostatin M (OM), all of which use gp130 as a signal transducer, significantly inhibited mouse placental lactogen-II (mPL-II) secretion by cultured placental cells from Days 7, 9, and 12 of pregnancy. These cytokines significantly stimulated mPL-I secretion by cells from Day 9, but not Day 7, of pregnancy. An antibody to LIF completely blocked the stimulatory and inhibitory effects of LIF on mPL-I and mPL-II secretion, respectively. LIF and OM decreased the abundance of mPL-II mRNA in placental cells. Double immunocytochemistry for mPL-I and mPL-II indicated that LIF, OM, and IL-11 significantly increased the number of giant cells containing only mPL-I or both mPL-I and mPL-II but decreased the number of giant cells containing only mPL-II. IL-6, which also uses gp130 as a signal transducer, inhibits mPL-II secretion only after midpregnancy; however, addition of soluble IL-6 receptor (sIL-6R) together with IL-6 resulted in a significant inhibition of mPL-II secretion before midpregnancy. Treatment of cells from Day 12 of pregnancy with IL-6 during the first 2 days of culture resulted in significant inhibition of mPL-II secretion by the third day of culture.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7492694 TI - A porcine homolog of human integrin alpha 6 is a differentiation antigen of granulosa cells. AB - To search for new molecules relating to the mechanism(s) of follicular differentiation, we raised a new monoclonal antibody (mAb), POG-2, which reacted with the surface of porcine ovarian granulosa cells. An immunohistological study revealed that the antigen appeared initially on the primary follicles. The antigen expression increased during follicular development, reached the highest level in small follicles 1-2 mm in diameter, and then decreased in the preovulatory follicles. The antigen was not detected on luteal cells in the CL. SDS-PAGE profiles of POG-2 antigen purified from isolated porcine granulosa cells by immunoaffinity chromatography indicated that it consisted of two proteins (120 and 130 kDa) and that the antigenic epitope of POG-2 was on the 120-kDa protein. We next purified a large amount of POG-2 antigen from the homogenate of porcine ovaries by immunoaffinity chromatography. Analysis of the partial amino acid sequence of the 120-kDa protein showed that 16 amino acids from the amino terminus were very similar to those of both human and murine mature integrin alpha 6, with 88% homology. SDS-PAGE profiles revealed that the 120-kDa protein was bound to a 28-30-kDa protein by a disulfide bond; this was compatible with the reported characteristics of human integrin alpha 6. We then examined the immunoreactive distribution of integrin alpha 6 using anti-integrin alpha 6 mAb GoH3. The staining profiles obtained with GoH3 were very similar to those obtained with POG-2. These findings showed that the molecule detected by POG-2 was a porcine homolog of human integrin alpha 6 and that integrin alpha 6 was a differentiation antigen of porcine granulosa cells, suggesting an involvement of integrins in follicular differentiation. PMID- 7492695 TI - Pulsatile gonadotropin-releasing hormone (GnRH) increases concentrations of GnRH receptor messenger ribonucleic acid and numbers of GnRH receptors during luteolysis in the ewe. AB - As circulating concentrations of progesterone decrease during the early preovulatory period, concentrations of mRNA encoding ovine GnRH receptor in the anterior pituitary gland increase. The purpose of this study was to determine whether removal of progesterone affects amounts of GnRH receptor mRNA directly or whether withdrawal of progesterone affects GnRH receptor gene expression indirectly by permitting secretion of GnRH to increase. Ovulation was induced in seasonally anestrous ewes, and luteolysis was initiated with prostaglandin F2 alpha (PGF2 alpha) 11 or 12 days later. Anterior pituitary glands were collected 0 h, 4 h, 12 h, or 24 h after treatment with PGF2 alpha, and 24 h after injection of saline (n = 3 or 4 animals/group). Two groups of ewes (n = 3) received infusions of GnRH (250 ng infused over 6 min) hourly for 12 h; luteolysis was induced in one of these groups at the time that treatment with GnRH was initiated, and anterior pituitary glands were collected at the end of the 12-h infusion period. Blood samples were collected at 15-min intervals for 12 h from all ewes treated with GnRH and from animals administered PGF2 alpha and killed 12 h later. No differences in concentrations of GnRH receptor mRNA, numbers of GnRH receptors, or circulating concentrations of progesterone or estradiol were detected between groups of animals at 0 h and 24 h after treatment with saline; therefore, data from these control groups were combined. Concentrations of progesterone in serum decreased in PGF2 alpha-treated ewes and were lower (p < 0.05) than those in controls 24 h after treatment with PGF2 alpha.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7492696 TI - Synchronous sperm penetration of zona-free mouse eggs in vitro. AB - To synchronize sperm penetration of zona-free eggs immediately after insemination, zona-free eggs preloaded with Hoechst-33342 were inseminated under various conditions. Insemination was mostly conducted at 10 sperm/microliter. In preliminary experiments, fatty acid-free BSA (FAF) was more satisfactory for sperm penetration than fraction V BSA, and FAF was used in the following experiments. Only 26% of zona-free eggs were fertilized at 10 min after insemination when the eggs were inseminated immediately after zona removal and preloading. However, egg preincubation significantly improved the penetration rate (1 h preincubation: 63%, 2 h preincubation: 82% penetrated 10 min after insemination). Some eggs preincubated for 2 h were already penetrated at 3 min (7%), and the rate gradually increased in a time-dependent manner (3 min: 7%, 5 min: 30%, 10 min: 80%). The rate further improved as the sperm concentration was increased; the maximal level was obtained at 160 sperm/microliters. At 160 sperm/microliters, 54% of the eggs were penetrated at 3 min and 78% were at 5 min. These results indicate that it is possible to synchronize the sperm entry and that not only sperm but zona-free eggs should be preincubated before insemination. These data are considered valuable for investigating early events in fertilization. PMID- 7492697 TI - Fertilization in vitro of mouse ova from inbred and outbred strains: complete preimplantation embryo development in glucose-supplemented KSOM. AB - A new medium derived from the use of sequential simplex optimization methods (SOM) that overcomes the block to development beyond two cells in vitro in embryos of the CF1-cultured strain of mouse has recently been described. Contrary to previous reports, glucose was shown to have no significant inhibiting effect on embryo development to the blastocyst stage in SOM. A modification of SOM, designated KSOM, with an increased concentration of Na+ (95 mM) and K+ (2.5 mM), which has been described elsewhere, also supports growth beyond the two-cell block. KSOM produces a higher rate of compaction, a larger yield of blastocysts, and an increased rate of cell division of the trophoblast cells. We have reexamined the glucose effect by varying the concentration of glucose (either 0.2 mM or 5.56 mM) in KSOM and determined the ability of these media to support preimplantation development of CF1 female x B6D2F1 male zygotes through the blastocyst stage. Glucose is shown to have no significant inhibiting effect on development to the blastocyst stage. The yield of blastocysts is typically 85% 90%. A modification of KSOM derived from this study, designated modified KSOM, with an increased concentration of glucose (5.56 mM) and supplemented with 4 mg ml-1 BSA is now shown to support high rates of fertilization in vitro of CF1 ova with hybrid B6D2F1/CrlBR sperm and subsequent development of zygotes beyond the two-cell stage to blastocysts in high yield.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7492698 TI - Pulsatile gonadotropin secretion determined by frequent sampling from the intercavernous sinus of the mare: possible modulatory role of progesterone during luteolysis. AB - Twelve horse mares were used in a repeated-measures design consisting of 3 replicates of 4 mares each. On Day 6 following ovulation, luteolysis was initiated with an i.m. injection of prostaglandin F2 alpha (PGF2 alpha; Lutalyse, 10 mg). Either 12 (-12) or 36 (-36) h before PGF2 alpha (PRE), blood samples were collected simultaneously from the intercavernous sinus (ICS) and jugular (JUG) vein at 10-min intervals for an 8-h period. Pituitary capacity to exogenous GnRH (2 micrograms/kg BW, i.v.) was evaluated at the alternate time point within this period. Frequent sampling and GnRH challenge were repeated 36 (+36) or 60 (+60) h after PGF2 alpha (POST). Progesterone reached a nadir 48 h after PGF2 alpha, while the first significant elevation in estradiol (p < 0.05) was not detected until 156 h. PRE LH and FSH pulse frequencies in ICS samples were 0.6 +/- 0.3 and 1.8 +/- 0.8 pulses/8 h, respectively. Three high-amplitude, concurrent ICS pulses of LH and FSH were observed during this period. These were associated with elevated gonadotropin levels in the corresponding JUG samples. In contrast, low amplitude ICS pulses at this time were predominately releases of FSH alone that were not detectable in the JUG. Following luteolysis, POST gonadotropin pulse frequencies in the ICS increased (p < 0.001) to 7.4 +/- 0.4 and 6.8 +/- 0.5 pulses/8 h for LH and FSH, respectively. Concurrent LH and FSH releases of relatively low amplitude characterized 87.7% of the POST ICS pulses, none of which were measurable in corresponding JUG samples.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7492699 TI - Dissociation of relaxin and progesterone secretion from the primate corpus luteum by acute administration of a 3 beta-hydroxysteroid dehydrogenase inhibitor during the menstrual cycle. AB - The factors regulating relaxin production by the primate CL during spontaneous menstrual cycles and in early pregnancy are poorly understood. Since the CL produces steroids, notably progesterone, and expresses progesterone receptors, luteal progesterone may act locally to regulate relaxin production. For the current study, either trilostane (600 mg daily)--a 3 beta-hydroxysteroid dehydrogenase inhibitor--or vehicle was administered to rhesus monkeys during the midluteal phase (Days 6 and 7) of spontaneous menstrual cycles to examine the effects of reduced luteal progesterone synthesis on relaxin secretion. Trilostane treatment reduced serum concentrations of progesterone within 3 h of initial administration and maintained low levels typical of the follicular phase (< 1 ng/ml), causing premature menses without significant alteration in serum bioactive LH levels. Nevertheless, the patterns and levels of circulating relaxin, as measured by homologous macaque ELISA, were not different between trilostane- and vehicle-treated monkeys, with relaxin levels peaking in both groups by Day 13 of the luteal phase. To determine if chorionic gonadotropin (CG) injections simulating early pregnancy could stimulate relaxin production in a progesterone-depleted environment, trilostane or vehicle was administered as described above, followed by injections in increasing dosages of human (h) CG beginning 3 days after initial trilostane administration. Serum progesterone levels in trilostane-treated animals were significantly reduced prior to and during hCG treatment when compared with vehicle-treated animals. However, serum relaxin levels were comparable between these groups; relaxin levels peaked approximately 10-fold above pre-hCG levels in both trilostane- and vehicle treated animals.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7492700 TI - Determination of the prorelaxin nucleotide sequence and expression of prorelaxin messenger ribonucleic acid in the golden hamster. AB - The objectives of this study were to determine the nucleotide and derived amino acid sequences of hamster prorelaxin and to evaluate prorelaxin mRNA expression during gestation. Total RNA was extracted from tissues through use of guanidinium hydrochloride methodology. A 23-bp oligonucleotide pool derived from the N terminal amino acid sequence of hamster prorelaxin and Day-14 placental RNA were used in 3' RACE methodology to generate a prorelaxin-specific cDNA. This cDNA fragment (940 bp) was utilized to screen a Day-16 hamster placental cDNA library, and a clone containing the entire coding region was identified. Nucleotide sequence analysis revealed a 531-bp open reading frame for preprorelaxin. The derived amino acid sequence contained a 22 amino acid signal peptide followed by a 155 amino acid prorelaxin sequence with a calculated molecular weight of 17,500. The derived prorelaxin amino acid sequence had 51.8%, 42.9%, 41.7%, and 38.2% homology with rat, human-1, human-2, and pig prorelaxins, respectively. Expression of prorelaxin mRNA during the latter half of gestation was evaluated by Northern and/or slot-blot analysis using the 940-bp cDNA fragment as a hybridization probe. Prorelaxin mRNA was first detected in the placenta on Day 8. Levels of the 900-bp transcript increased to a plateau on Days 10, 12, and 14 and subsequently increased further on Day 15. Prorelaxin mRNA was not detected in ovary, hippocampus, or neocortex. PMID- 7492702 TI - Infectious hazards associated with blood transfusion. PMID- 7492701 TI - Oral immunization with attenuated Salmonella expressing human sperm antigen induces antibodies in serum and the reproductive tract. AB - Induction of immune responses in the reproductive tract will be crucial for a functional gamete antigen-based antifertility vaccine. Here we describe the construction and development of an avirulent Salmonella as an oral vaccine delivery vector to elicit sperm-specific immune responses in reproductive tract secretions. A cDNA sequence encoding the human sperm antigen SP10 was cloned on an asd+vector and expressed to a high level in an avirulent delta cya, delta crp, and delta asd vaccine strain of Salmonella typhimurium. Oral immunization of female BALB/c mice with this recombinant Salmonella elicited high-titer anti-SP10 IgG antibodies in serum and IgA antibodies in vaginal secretions. Anti-SP10 antibody titers could be increased by secondary and tertiary oral administrations of the recombinant Salmonella. Induction of sperm-specific antibodies in the reproductive tract following oral administration of a recombinant Salmonella could lead to the development of a simple, safe, efficient, and easy-to-use antifertility vaccine. PMID- 7492703 TI - Medication-related adverse reactions and the elderly: a literature review. PMID- 7492704 TI - New bioactivation mode for vascular prostheses made of Dacron polyester. AB - The performance of vascular prostheses, especially those of small calibre, still presents several problems. The use of polyesters such as the well-known poly(ethylene terephthalate) (Dacron) and, more recently, poly(tetrafluoroethylene) and polyurethanes provides a marked improvement. We have developed a process to increase the efficiency of such prostheses, using the ability of the Dacron polyester to be covalently modified by proteins under mild chemical conditions. The biomaterial is obtained from the particular interaction of elastin-solubilized peptides with type I + III collagens under very precise conditions. We have obtained an irreversible covalent bonding on Dacron. These biological conditions and the addition of connective proteins and glycosaminoglycans conferred on the resulting material a composition and a three dimensional structure equivalent to the subendothelium, providing bioactive properties suitable for the development of a vascular prosthesis. PMID- 7492705 TI - Permeability and biocompatibility of a new hydrogel used for encapsulation of hepatocytes. AB - A new high-water-content (83%) and highly permeable anionic polyelectrolyte hydrogel was obtained by phase inversion of a polymer solution containing 6% polyacrylonitrile-sodium methallylsulphonate, 91% dimethylsulphoxide and 3% physiological saline solution. Hydrogel-based hollow fibres (HFs) were fabricated with a co-extrusion apparatus in collaboration with Hospal (France). HFs have an internal diameter of 800 microns and a wall thickness of 100 microns. Experimental results demonstrated that hydrogel-based HFs were permeable to albumin (mol. wt 69,000) and human immunoglobulin G (150,000), but were impermeable to immunoglobulins A (170,000) and M (900,000) after 24 h of diffusion. In vitro, the viability of isolated rat hepatocytes injected into the HFs was 64 +/- 6% after 10 d versus 30 +/- 5% for hepatocytes cultured in Petri dishes (P = 0.0001). Under these conditions, the amount of albumin released by encapsulated hepatocytes was 12 +/- 3 micrograms/24 h/10(6) cells at day 10, whereas at that time no albumin was released by hepatocytes cultured in Petri dishes. In vivo, histological study of hydrogel HFs implanted up to 6 wk in the peritoneum of rats revealed a low inflammatory tissue reaction without giant multinucleate cells in the foreign tissue, which decreased after the third week. The survival rate of encapsulated hepatocytes was over 85% 45 d after transplantation in the peritoneum of syngeneic Lewis rats. Therefore, this hydrogel demonstrates highly favourable properties for encapsulation of hepatocytes with regard to its biocompatibility, permeability and ability to maintain hepatocytes in a functional state for prolonged periods. PMID- 7492706 TI - Covalent immobilization of alpha-amylase onto pHEMA microspheres: preparation and application to fixed bed reactor. AB - Microspheres of poly(2-hydroxyethyl methacrylate) with and without cross-linker were prepared by suspension polymerization. As the amount of cross-linker increased, the equilibrium water content, enzyme loading, immobilization efficiency and recovered activity were all adversely affected. Enzyme alpha amylase was immobilized onto the microspheres after activation with epichlorohydrin. The Km value for the immobilized enzyme (0.90% w/v) was much greater than that of the free enzyme (0.53% w/v). It was found that the inactivation constant (ki) increased from 2.23 x 10(-8) min-1 at 20 degrees C to 1.45 x 10(-4) min-1 at 60 degrees C. Since the enzyme activity increased as the temperature increased, the temperature profile yielded a peak at 50 degrees C. For free enzyme this is at 45 degrees C. The residence time was proportional to the percentage hydrolysis until a residence time of 12 min was reached. Beyond this the activity increase could not match the increase in residence time. The pH profile yielded a broadening upon immobilization in addition to a small shift to higher pH (from 5.5 to 6.0). The continuous run at 30 degrees C, 1.0% w/v starch concentration and flow rate of 40 cm3 h-1 led to only 20% loss in activity after a 120 h operation. PMID- 7492707 TI - Glutaraldehyde cross-linked chitosan microspheres as a long acting biodegradable drug delivery vehicle: studies on the in vitro release of mitoxantrone and in vivo degradation of microspheres in rat muscle. AB - Chitosan microspheres were prepared from 74% deacetylated chitin by the glutaraldehyde cross-linking of an aqueous acetic acid dispersion of chitosan in a mixture of liquid paraffin and petroleum ether stabilized using sorbitan sesquioleate as the surfactant. Cross-linking and hardening of the spherical particles were achieved by the addition of glutaraldehyde-saturated toluene through the organic phase. A relatively novel antineoplastic agent, mitoxantrone, was incorporated into the microspheres and the drug release was studied in vitro into phosphate buffer for over 4 weeks at 27 degrees C. Drug release was found to be effectively controlled by the extent of cross-linking. Only about 25% of the incorporated drug was released over 36 days from microspheres of high cross linking density. Implantation of placebo chitosan microspheres in the skeletal muscle of rats was carried out in order to assess the biocompatibility and biodegradability of the microspheres. Histological analysis showed that the microspheres were well tolerated by the living tissue. However, no significant biodegradation of the material was noticed over a period of 3 months in the skeletal muscle of rats. Data obtained indicate the possibility of using cross linked chitosan microspheres as a drug carrier for sustained drug release for very long periods. PMID- 7492708 TI - Pore strain behaviour of collagen-glycosaminoglycan analogues of extracellular matrix. AB - Mechanical properties of a highly porous collagen-glycosaminoglycan (CG) analogue of extracellular matrix were investigated. The stress-strain behaviour of the matrices displayed the typical concave-up shape characteristic of porous materials and soft tissues. Deformation behaviour of the material was further investigated using novel methodology to determine material strains at the micrometre level. Central to the methodology was the use of scanning electron microscopy (SEM) in conjunction with a digital image analyser. The method for strain determination at the micrometre level was verified by comparison with measurement of surface strain using an optical method. An unexpected finding in these materials is that Poisson's ratio varies with engineering strain. A geometrical/mathematical model was developed to explain this behaviour. These results encourage further investigation of the mechanical properties of these materials, in order to provide important insights into the physical microenvironment in which cells reside. PMID- 7492709 TI - Viscoelastic and thermal properties of collagen/poly(vinyl alcohol) blends. AB - Blends of poly(vinyl alcohol) (PVA) with collagen and gelatin, prepared from aqueous solution by solvent casting, were investigated by differential scanning calorimetry (DSC) and dynamic mechanical thermal analysis (DMTA). After conditioning at 51% relative humidity, collagen and PVA show nearly coincident glass transition temperatures (Tg approximately 35 degrees C), while gelatin has a higher Tg (approximately 70 degrees C). Gelatin/PVA blends show two invariant Tgs, whose temperature and associated specific heat increment clearly indicate the coexistence of two amorphous phases composed of the pure components. Owing to similarity of the Tgs of collagen and PVA after humidity conditioning, DSC offers no indication on miscibility of collagen/PVA blends. In DMTA experiments, where absorbed water freely evaporates from the samples during the thermal scan, PVA shows a glass transition relaxation at about 50 degrees C, while both gelatin and collagen display an intense glass transition in the vicinity of 230 degrees C. The DMTA spectra of collagen/PVA and gelatin/PVA blends show two invariant glass transition relaxations at about 50 and 230 degrees C. Absence of any Tg shift with composition demonstrates that the blend components are immiscible. However, blends of PVA with collagen and gelatin form optically clear films with good mechanical properties over the whole range of compositions. It is found that at T > Tg (PVA) the elastic modulus (E') of the blends strongly increases with increasing content of the biopolymer. In the case of collagen/PVA blends, experimental E' values agree with the predictions of a simple two-phase composite model with phases connected in parallel. It is concluded that, though thermodynamically immiscible with both native and denatured collagen, PVA forms mechanically compatible blends with collagen and gelatin. PMID- 7492710 TI - Mechanical properties and histological evaluation of sintered beta-Ca2P2O7 with Na4P2O7.10H2O addition. AB - The ultimate goal of implantation of biomaterials in the skeleton is to reach full integration of the non-living implant with the living bone. The biomaterial can be used much as a bone graft, resorbing or dissolving as bone growth occurs, and the end result is a new remoulded bone. Calcium pyrophosphate, Ca2P2O7, is one of the intermediate products of bone mineralization. beta-Dicalcium pyrophosphate (beta-DCP) doped with certain amounts of Na4P2O7.10H2O was prepared as the developed material. Na4P2O7.10H2O was used as a liquid-phase additive to improve the sintering process and promote physiological bioresorbability. Compressive strength and four-point bending strength were measured by the Bionix test system 858. The mechanical strength of the sintered beta-DCP increased with the addition of Na4P2O7.10H2O up to 5 wt%, but thereafter decreased. The microstructure and crystal structure were analysed by the techniques of SEM, EPMA, TEM and XRD. The relationship between the mechanical strength of the sintered bioceramics and the Na4P2O7.10H2O dopant was examined in terms of the presence of NaCa(PO3)3, grain growth and abnormal grain coalescence while the dopant increased. Preliminary in vivo evaluation was studied by rabbit femur condyle implantation. There was no inflammation or any toxic sign during the experimental period. The histological section of intraosseous implantation revealed that the new bone deposited directly on the surface of the material in the fourth week after operation. The implant gradually decreased in volume and was replaced by the surrounding regenerated bone in the rabbit condyle in vivo environment. The results led us to conclude that the developed material has great potential as a biodegradable bone substitute. PMID- 7492711 TI - Shielding of augmented tendon--tendon repair. AB - Strength and function of autogenic and xenogenic reconstruction of digital extensor tendons was examined in an ovine model. In this study, tendon-graft junctions were formed by either suture augmented with a woven polyester tube (A), or augmented and shielded from surrounding tissues by chemically-treated bovine pericardium (S). By 12 wk, both A and S sheep had returned to full range of motion. Mechanical strength of both the autograft-host and xenograft-host repair sites was similar, with a pooled strength of 131 +/- 25 N (n = 15). Similarly, the mid-portion xenograft strengths were constant at approximately 366 +/- 97 N (n = 7). In contrast, mid-portion autograft strengths decreased from 380 +/- 110 N (N = 4) to 120 +/- 66 N (n = 4) if shielding was omitted. The loss in autograft strength was attributed to loss of function associated with adhesions. The use of the augmentation device coupled with an adhesion barrier gives higher initial reconstruction strength and improved function during the host repair period up to 12 wk. PMID- 7492712 TI - Chondrocyte behaviour within different types of collagen gel in vitro. AB - In cartilage repair experiments chondrocytes are transplanted into osteochondral defects. Biological substances are used as cell vehicles and are likely to play an important role in the outcome of these studies. Collagen gel is formed by polymerization of type I collagen and is used in plastic surgery and for three dimensional culture systems. To test collagen gel as a potential vehicle for transplantation, we evaluated chondrocyte behaviour in vitro in different collagen gels. Collagen type I was extracted and purified from rat tail tendon and fetal calf skin and compared with commercially available collagen type I. After suspension of bovine chondrocytes, five different collagen gels were cultured for 14 days and evaluated by light and electron microscopy. Cells proliferated within all gels and synthesized proteoglycans as assessed by 35S incorporation; 40-90% of cells maintained a chondrocyte-like morphology after 1 week in culture depending on the type of collagen gel. Synthetic and secretory activity was confirmed by electron microscopy. Based on these results, calf skin collagen is recommended for culturing chondrocytes for implantation. PMID- 7492713 TI - Bone-bonding behaviour of poly(ethylene oxide)-polybutylene terephthalate copolymer coatings and bulk implants: a comparative study. AB - A range of poly(ethylene oxide)-polybutylene terephthalate (PEO-PBT) copolymers (70-30% PEO), both as coating on titanium alloy as well as bulk cylinders, was press-fit implanted in the diaphyseal femur of 16 goats. At early survival times (4 wk), a high degree of cortical bone contact was observed for bulk implants using light microscopy and this was confirmed by backscatter electron microscopy. This was attributed to the swelling behaviour of PEO-PBT copolymers. At this stage, bone contact was also revealed for coated implants, but to a lesser extent. At a later stage (12 wk), bone bonding was demonstrated both morphologically and by X-ray microanalysis, at the interface of 70:30 PEO-PBT bulk as well as 70:30 PEO-PBT-coated implants. Bone bonding increased with time (26 and 52 wk) for this PEO-PBT proportion and was also observed for 60:40 and 55:45 implants, although less frequently. For 40:60 and 30:70 PEO-PBT proportions, bone bonding was not shown. Based on these qualitative data, it was not possible to differentiate between coated and bulk implants with respect to bone bonding. This study demonstrated that the application of PEO-PBT elastomers as coatings does not alter the bone-bonding properties. It was therefore concluded that PEO-PBT coatings are beneficial over the bone-bonding but brittle ceramic coatings, due to their flexibility. In addition, the bone-bonding capacities of these PEO-PBT coatings surpass the non-bonding behaviour of currently available flexible coatings. PMID- 7492714 TI - Bioactive glass coating: physicochemical aspects and biological findings. AB - Glass coating material was investigated before and after spraying to see whether it maintained the chemical and physical properties; in vivo and in vitro studies were done to evaluate the biological results. Following the spraying process, the Biovetro coating on the TiAl6V4 plate--as evidenced from chemical and physical analysis--maintains the properties of the original glass unchanged as far as the amorphous structure and its behaviour in a hydrolytic environment are concerned. In vitro and in vivo studies underline the positive features of the coating obtained by the plasma spray process, confirming that it has the typical properties of bioactive glass patented under the trade mark, Biovetro, i.e. biodegradability and osteoconductivity already confirmed by previous experimental protocols carried out by our group using powdered and fibre Biovetro. PMID- 7492715 TI - Surface modifications of glass-reinforced hydroxyapatite composites. AB - Surface modifications of glass-reinforced hydroxyapatite composites immersed in a simulated physiological solution were studied using X-ray photoelectron spectroscopy, scanning electron microscopy and energy dispersive X-ray spectroscopy. In the first stages of the apatite formation process, calcium and silicon ions were leached out from the surface of the composites. After 12 days of immersion, apatite crystals were detected on the surface indicating bioactive behaviour. Sodium ions attributed to sodium sulphate were also found in the apatite layer. Bound water on the surface also increased with immersion time. PMID- 7492716 TI - Non-decay type fast-setting calcium phosphate cement: composite with sodium alginate. AB - Non-decay type fast-setting calcium phosphate cement (nd-FSCPC) was prepared by introducing sodium alginate (0-2.0 wt%) into the liquid phase of FSCPC. nd-FSCPC was stable even when the cement paste was immersed in distilled water immediately after mixing, whereas conventional FSCPC (c-FSCPC) decayed completely within 1 min upon immersion. The setting time of the cement, approximately 5 min, was not dependent on the presence of sodium alginate. In contrast, the introduction of sodium alginate into conventional CPC, i.e. CPC without neutral phosphate in the liquid phase, resulted in no setting when the amount of sodium alginate introduced was more than 1 wt%. Powder X-ray diffraction analysis revealed no significant difference for the conversion of cement to apatite for any concentrations of sodium alginate studied (0-2.0 wt%). The mechanical strength of the cement increased rapidly with the addition of sodium alginate up to 0.8 wt% when the cement paste was immersed and kept in distilled water at 37 degrees C, whereas further addition of sodium alginate decreased the mechanical strength. The results obtained in this investigation, taken together with sodium alginate's known excellent biocompatibility and absorption behaviour, indicate that the use of sodium alginate composite FSCPC as nd-FSCPC should be of value in orthodontics and oral and maxillofacial surgery where the cement is exposed to blood. PMID- 7492717 TI - Pullout strength of fixation screws from polymethylmethacrylate bone cement. AB - Polymethylmethacrylate bone cement is often used to fill voids and increase the strength of osteoporotic and pathological bone. However, it is unclear as to which method of cement augmentation provides optimal screw fixation. This study was conducted to determine which of the current cement augmentation techniques provides the strongest construct when used in association with orthopaedic fixation screws. Pullout strength was determined for screws placed in sawbones with no cement, soft cement, doughy cement and hard cement after drilling and tapping. All cement-screw constructs were significantly stronger than the no cement group. Screws placed in doughy cement had a significantly higher pullout force than those placed in hard cement. Pullout strength of screws placed in soft cement was intermediate between the other cement techniques but not significantly different from either group. PMID- 7492718 TI - Do porous calcium hydroxyapatite ceramics cause porosis in bone? A bone densitometry and biomechanical study on cortical bones of rabbits. AB - Porous calcium hydroxyapatite (CHA) ceramics are biocompatible and present osteoconductive properties. These ceramics are widely used in orthopaedic surgery; however, it is not yet known whether they have some adverse effects on bone and bone marrow healing. Our previous radiological study revealed possible local porosis at the adjacent sites of the CHA ceramic. Histological findings of the same study revealed bone marrow swelling and depletion at the implantation site. Osteoclasts removed particles of the implant that may be the cause of local porosis. In the present study, possible local osteoporosis was evaluated by bone densitometry analyses, and compression and three-point bending tests. CHA particles were implanted into the left limbs and a sham operation was utilized on the right limbs of 75 white rabbits. The animals were followed up for 23 weeks for bone mineral density and for 6 months for biomechanical analyses. The CHA implanted area and its distal or proximal adjacent areas were evaluated with a Hologic QDR-2000 bone densitometer. Three-point bending and compression tests were performed with an M-30 K material testing device. The results revealed a time-dependent bone density increase at the CHA implantation site and no significant porosis at adjacent areas of the implant. The stiffness of CHA implanted bones in three-point bending is larger than that of the control group. CHA-implanted rabbit bones presented a different fracture pattern from the control group. The stiffness of the control and CHA-implanted bones generally increased with time indicating no adverse effects of porous CHA ceramics in bone and bone marrow healing. The clinical relevance of this work is that porous CHA ceramics do not cause local porosis at adjacent areas when implanted into osseous sites.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7492719 TI - Effects of selected thermal variables on the mechanical properties of trabecular bone. AB - Osteoarticular allografts are commonly used in the treatment of segmental bone loss due to a wide resection of tumour. While the use of such grafts has met with considerable clinical success, fractures are a recognized complication of allograft use. Although trabecular bone can play an important structural role in the function of segmental allografts, few data exist on the effects of common storage and sterilization procedures on the mechanical properties of trabecular bone. To this end, we investigated with these experiments the effects of freezing at -20 degrees C, freezing at -70 degrees C, eight freeze-thaw cycles at -20 degrees C, freeze-drying, boiling and autoclaving on the compressive modulus and strength of bovine trabecular bone. Of these treatments, boiling and autoclaving were the only treatments to alter the properties of bovine trabecular bone, resulting in 26 and 58% reductions in strength, respectively. Autoclaving also significantly reduced the compressive modulus by 59%. From these data, freezing at temperatures between -20 and -70 degrees C does not appear to compromise the structural integrity of trabecular bone. PMID- 7492720 TI - Compaction assay: a rapid and simple in vitro method to assess the responsiveness of a biopolymer matrix to enzymatic modification. AB - A rapid and simple in vitro method is described which measures the extent of unrecoiled solids compression when a complex biopolymer is subjected to a centrifugal force. This method, termed the compaction assay, was used to assess the response of purulent cystic fibrosis (CF) sputum samples to the addition of recombinant human deoxyribonuclease I (rhDNase). Enzyme treatment resulted in a dramatic decrease in DNA size, a redistribution of total DNA content from the pellet to supernatant, a significant decrease in that pellet volume and a decrease in elastic modulus. Sample elasticity, measured by a dynamic cone and plate viscometer, could be related to compaction assay results. These results suggest that the compaction assay may be a useful in vitro method for rapidly assessing the actions of enzymatic disruption of a complex biopolymer, such as that observed for the actions of rhDNase on purulent airway secretions. PMID- 7492721 TI - Swelling/deswelling of anionic copolymer gels. AB - Studies of dynamic and equilibrium swelling of ionic gels are important in understanding the diffusion of physiologically important fluids in materials for site-specific controlled drug delivery applications. The dynamic and equilibrium swelling properties of dry glassy poly(2-hydroxyethyl methacrylate-co-methacrylic acid) and poly(2-hydroxyethyl methacrylate-co-acrylic acid) polymeric networks were studied as a function of pH, ionic strength, nature of the counterion and buffer composition. The mechanism of water diffusion in these gels became more anomalous as the pH of the swelling medium increased and as the ionic strength decreased at a constant pH > or = pKa,gel. The mechanism of water diffusion was Fickian in all unbuffered swelling media at pH 4.0, which is lower than the pKa,gel. The pKa,gel of these gels was between 5.5 and 6. At pH 4.0, the diffusion mechanism was independent of ionic strength. This swelling behaviour is explained in terms of the concept of ion osmotic swelling pressure and ion exchange kinetics. PMID- 7492722 TI - Blends of synthetic and natural polymers as drug delivery systems for growth hormone. AB - In order to overcome the biological deficiencies of synthetic polymers and to enhance the mechanical characteristics of natural polymers, two synthetic polymers, poly(vinyl alcohol) (PVA) and poly(acrylic acid) (PAA) were blended, in different ratios, with two biological polymers, collagen (C) and hyaluronic acid (HA). These blends were used to prepare films, sponges and hydrogels which were loaded with growth hormone (GH) to investigate their potential use as drug delivery systems. The GH release was monitored in vitro using a specific enzyme linked immunosorbent assay. The results show that GH can be released from HA/PAA sponges and from HA/PVA and C/PVA hydrogels. The initial GH concentration used for sample loading affected the total quantity of GH released but not the pattern of release. The rate and quantity of GH released was significantly dependent on the HA or C content of the polymers. PMID- 7492723 TI - In vitro and in vivo studies of the effect of the concentration of plasticizer di(2-ethylhexyl) phthalate on the blood compatibility of plasticized poly(vinyl chloride) drain tubes. AB - Evaluation of the effect of the plasticizer di(2-ethylhexyl) phthalate (DEHP) has been carried out on the properties of blood compatibility of plasticized poly(vinyl chloride) (PVC) through an in vitro registering of the amount of total protein in relation to the amount of DEHP. The influence of DEHP on the properties of blood compatibility has also been evaluated in vivo by tracing through a modified method of Gott (LV Gott and A Feruse, Bull NY Acad Med 1972; 48(2): 482-491) the presence of a thrombus and the patency of tubes made from the material tested, as well as the patency of tubes made from the same material having surfaces coated with paraffin and complexone. For all implanted samples the changes in coagulation time and the amount of haemoglobin after their stay inside a blood vessel in the abdominal cavity of a dog have been registered. PMID- 7492724 TI - Amalgams made from liquid Hg-In. AB - The addition of indium to amalgam appears to reduce the mercury vapour above fillings during setting. In the present work the structure and the physical properties of amalgams with In additions to the mercury up to 30 vol.% have been investigated. No new phase or undissolved in could be detected by X-ray diffraction and scanning electron microscopy studies. Energy dispersive spectroscopy measured 4 at.% In in the gamma 1-phase of an amalgam with 20 vol.% In added to the mercury. At the same time, the Ag/Hg ratio was substantially increased compared with amalgam having no In addition. The compression strength decreased with increasing In content, and for amalgam with as much as 30 vol.% In added to the mercury, it fell below the minimum requirement of ISO 1559 (Alloys for dental amalgam, International Organization for Standardization, Geneva, 1986). An abrupt drop of the load at the onset of failure and cracks running through the gamma 1-phase, as well as the eta-phase, indicates embrittlement of these phases with increasing In content. PMID- 7492725 TI - [Biomechanics of healing of cortical autotransplants after intramedullary nailing and device removal]. AB - The aim of the study was to investigate the biomechanical properties of segmental cortical autografts subjected to extracorporeal treatment. For this purpose, 12 beagle dogs were submitted to tibial osteotomy, when 25 mm-long diaphyseal segments of bone were removed and treated outside the body by autoclaving or gamma radiation (Co, 5 kGy). Subsequently, osteosynthesis involving the use of medullary nailing was performed. After 36 weeks, the dogs were killed painlessly and, after removing the nails, the tibias were tested for the biomechanical properties. In comparison with the contralateral tibia, a mean stiffness of 47% and 32% was measured for tibia treated by irradiation and autoclaving, respectively, while bending showed an increase of up to 590%. These results show the need to allow plenty of time for healing in the case of weightbearing transplants. When bealing is undisturbed, no biomechanical differences are found between proximal and distal osteotomies, although in the X-ray image the osteotomy gap (fracture line) remains visible, and scintigraphic activity persists for a longer period. This means that the radiological aspect of the former does not necessarily reflect actual biomechanical strength. PMID- 7492727 TI - [A new ultrasound method for examination of the breast]. AB - In the present paper, a new sonographic method of examining the breast is described. In the form of a laboratory model, it has been used to investigate 245 preoperative patients and 146 women referred for conventional sonography of the breast. All the patients had also undergone mammography. With the new procedure the examination situation is identical to that used for mammography, with a stainless steel plate (reference reflector) placed beneath the breast, and which appears on the B scan behind healthy tissue as a straight white line. Disordered reflectivity and elevation of this line indicate the presence of a lesion. Measurement of the lesion and the elevation of the reference line permit differentiation between benign and malignant disease. The use of both X-ray and sonographic findings improved the detection of cancers and reduced false-positive benign findings. Further technical improvements to this sonographic procedure could make it as rapid and reliable as mammography. PMID- 7492726 TI - [Cerebral monitoring in anesthesia and intensive care]. AB - To ensure efficient computer-based cerebral monitoring in the ICU and during anaesthesia, sophisticated and sometimes expensive equipment is needed. It is thus all the more important that the clinician should have a good understanding and knowledge of the various methods involved, the origins of the different signals and their relationships, and of the possibilities of artefacts occurring, if the information that this equipment is capable of providing is to be fully and properly utilized. The present paper describes a number of new techniques of cerebral monitoring, and their successful application in the intensive care unit and the operating theatre. PMID- 7492728 TI - [Comparison of the effectiveness of sodium azide and merthiolate as bacteriocidal agents--a NMR study]. AB - The pseudohalogenide sodium azide (NaN3) and the organometallic mercury compound merthiolate (sodium salt of ethyl mercurithiosalicylate) are commonly used as preserving solutions for biomaterials. In the present paper, these preserving solutions were compared for their efficacy in preserving porcine articular cartilage. For this purpose, porcine articular cartilage was incubated with phosphate buffer containing the corresponding amounts of NaN3 or merthiolate. Supernatants were assayed using NMR spectroscopy, and the content of soluble saccharides was determined using the dye Alcian blue. The results obtained clearly indicate that merthiolate is more effective than sodium azide. This makes the former a more suitable preserving agent for the storage of cartilage. A significant decrease in the lactate concentration, and enhancement of the acetate and the N-acetyl resonances indicate degenerative changes in the cartilage. However, when a large excess of sodium azide is used, its efficacy approaches that of merthiolate. PMID- 7492730 TI - Thiol oxidation by 1,2,3-oxadiazolinium ions, presumed carcinogens. AB - 3-Alkyl-1,2,3-oxadiazolinium ions 1 have been proposed as reactive intermediates in the activation of (2-hydroxyethyl)nitrosamines. The reaction of 3-methyl-1,2,3 oxadiazolinium tosylate (1a), 2-ethyl-1-methoxy-2-phenyldiazenium tetrafluoroborate (3), and 3-phenyl-1,2,3-oxadiazolinium triflate (1b) with thiols was investigated to determine the behavior of these compounds toward typical "cellular nucleophiles". Each of these substances oxidizes benzenethiol to diphenyl disulfide. The reaction in aqueous buffer at pH 7.4 is rapid. Reaction of 1b with benzenethiol gives, in addition to the disulfide, benzene, biphenyl, azobenzene, diphenyl sulfide, aniline, and glycolaldehyde. Similar products are obtained from 3. Phenyldiazene is postulated as an intermediate in this process, and its generation from phenyldiazoformate in the presence of benzenethiol gives similar products. Diazenes are presumed to arise by proton abstraction from the CH adjacent to N. The kinetics of reaction of 1a with N acetylcysteine to give the corresponding disulfide show first order dependence on each reactant and base catalysis. The data from these model chemical experiments suggest that 1,2,3-oxadiazolinium ions could react with abundant thiols in cells to lead to either their detoxification or radical processes emanating from diazenes. The occurrence of thiol-oxadiazolinium ion redox transformations could modulate the alkylation chemistry of these substances as well. PMID- 7492733 TI - Reaction of mucochloric acid with adenosine: formation of 8-(N6 adenosinyl)ethenoadenosine derivatives. AB - The bacterial mutagen mucochloric acid was reacted with adenosine in aqueous solutions at 37 degrees C. In the HPLC chromatograms of the reaction mixtures two peaks of unidentified products were observed at longer retention times than the previously characterized "etheno" and "ethenocarbaldehyde" adducts. Following isolation and purification with chromatographic methods, the products were characterized by UV absorbance, 1H and 13C NMR spectroscopy, and thermospray mass spectrometry. The products were found to consist of ethenoadenosine derivatives which bonded an additional adenosine unit to C-8 in the etheno bridge. In one of the products a formyl group and in the other an oxalo group was localized at C-7 in the etheno bridge. The yield of the products was about 0.04 mol% (calculated from the original amount of adenosine) in the reaction mixture held for 4 days at pH 7.4. It was concluded that mucochloric acid acts as an oxidative agent during the course of formation of the products. PMID- 7492729 TI - [In vitro studies of the specifications for methodological and technical equipment requirements of endoscopic infrared diaphanoscopy]. AB - Endoscopic infrared diaphanoscopy is a method of improving orientation in the nasal cavity during endoscopy-controlled endonasal surgery. The transillumination is expected to facilitate the localisation of the natural passageways between the paranasal sinuses and the nasal cavity during endoscopic procedures. The present study specifies the necessary conditions on the basis of in vitro examinations using an animal model (sheep) and an anatomical preparation of half of the human head. The questions as to what paranasal sinus is at all accessible to diaphanoscopy, whether the method actually enables the connecting pathways between an individual sinus and the nasal cavity to be identified, and what technical demands need to be met, are considered and answered. PMID- 7492732 TI - Role of compartmentation in promoting toxicity of leukocyte-generated strong oxidants. AB - A rigorous mathematical model is developed to describe the distribution of respiration-generated oxidants among reactive sites within the phagolysosomes of leukocytic cells. Reaction parameters include the diffusion coefficient of the oxidant, the intrinsic rate constants for its reaction with the phagosomal membrane and the cell envelopes of entrapped bacteria, the overall rate constant for its reaction with solution components of the phagosomal fluid, and the phagosomal dimensions. The model is used to describe the dynamics of randomly generated .OH and HCO3. radicals within the phagosome. These radicals were chosen because the necessary rate parameters either have been measured or could be reasonably estimated. The calculations show that .OH radical cannot be an effective bactericide unless generated in the immediate vicinity of the bacterial surface because its extreme reactivity precludes any significant diffusion. The HCO3. radical, however, is predicted to be a very effective toxin, even when relatively high concentrations of oxidant scavengers are present in the phagosomal fluid. In the absence of complicating features, the reactivity patterns of other less reactive oxidants (e.g., metal oxo or peroxo complex ions) are predicted to be very similar, although quantitative analysis is precluded by the lack of relevant rate data. For these oxidants, the predicted intraphagosomal toxicities differ markedly from toxicities measured in dilute bacterial suspensions because differences in mean diffusion lengths of the oxidants are unimportant in environments with the dimensions of phagosomes, but very important under in vitro conditions. The model is general and can be applied to other reactions occurring in similar microheterogeneous cellular and subcellular environments. PMID- 7492731 TI - Solution structure of an oligodeoxynucleotide containing the human n-ras codon 61 sequence refined from 1H NMR using molecular dynamics restrained by nuclear Overhauser effects. AB - The solution structure of the ras61 oligodeoxynucleotide duplex d(CGGACAAGAAG). d(CTTCTTGTCCG), which consists of codons 60, 61 (underlined), and 62 of the human n-ras protooncogene, was refined from 1H NMR data. The sequence contains a run of purines in the coding strand, with one R-Y step, A4.T19-->C5.G18, and one Y-R step, C5.G18-->A6.T17 (excluding the 5'-terminal base pair). The NMR data were consistent with a B-like helix as judged by characteristic internucleotide NOEs. The NOE intensities between purine H8 and purine anomeric protons were small as compared to the intensities between cytosine H5 and H6 protons, indicative of glycosyl torsion angles in the anti range. Cross-peaks were observed between purine H8 and pyrimidine H5 and CH3 protons on adjacent bases in the direction of purine (5'-->3') pyrimidine, but not in the direction pyrimidine (5'-->3') purine. Watson-Crick hydrogen bonding was intact and enabled the assignment of the exchangeable protons. A total of 226 experimental distance restraints were obtained. A restrained molecular dynamics and simulated annealing approach was utilized in the refinement. The data for 5 emergent molecular dynamics (MD) structures calculated from a B-form starting structure and 5 emergent MD structures calculated from an A-form starting structure refined to an average pairwise root-mean-square (rms) difference of 1.2 A, with maximum pairwise rmsd of 1.7 A. The accuracy of the emergent structures was assessed by complete relaxation matrix back-calculation. The sixth root residual index of 9.4 x 10(-2) was measured between the refined structures and the NOE data, suggesting that the former were in reasonable agreement with the data. The refined structures revealed an increased roll angle of 7 degrees in the codon 61 sequence at base step C5.G18-->A6.T17, which relieved the purine-purine clash in the minor groove, and in turn relieved the purine-purine clash in major groove between A4.T19 and C5.G18. A 3.7 A rise between C5.G18 and A6.T17 was calculated, which assisted in relieving the purine-purine clash. The local variations in the B-like conformation did not confer large structural alterations upon the ras61 sequence, but could be important in modulating the reactivity of the first as compared to the second adenine in codon 61. PMID- 7492734 TI - Three-dimensional quantitative structure-activity relationships of dioxins and dioxin-like compounds: model validation and Ah receptor characterization. AB - In the present study we have utilized comparative molecular field analysis (CoMFA), a three-dimensional quantitative structure-activity relationship paradigm, to explore the physico-chemical requirements for binding to the Ah (dioxin) receptor. Recent developments by Gillner et al. [(1993) Mol. Pharmacol. 44, 336-345] prompted us to review and revise our previous CoMFA/QSAR model [Waller, C. L., and McKinney, J. D. (1992) J. Med. Chem. 36, 3660-3666] to include a structurally-diverse training set of Ah receptor ligands ranging in size from naphthalene to indolo[3,2-b]carbazole nuclei. An exhaustive validation process utilizing external test sets and hierarchical cluster analysis routines was employed during model construction and is discussed herein. The limitations of the approach presented herein are discussed with respect to predictive ability of the CoMFA/QSAR models, which is demonstrated to be dependent on a balance between structural diversity and redundancy in the molecules comprising the training set. The results of our modified CoMFA/QSAR model are consistent with and unify all previously established structure-activity relationships established for less structurally-diverse training sets of Ah receptor ligands. As a result of the more complete nature of the series of molecules under examination in the present study, the CoMFA/QSAR steric and electrostatic field contour plots as well as the essential and excluded volume plots provide for a more detailed characterization of the molecular binding domain of the Ah receptor. The implications of the CoMFA/QSAR model presented herein are explored with respect to quantitative hazard identification of potential toxicants. PMID- 7492735 TI - Reaction between peroxynitrite and hydrogen peroxide: formation of oxygen and slowing of peroxynitrite decomposition. AB - Peroxynitrite, the reaction product of nitric oxide and superoxide, is a potent and versatile oxidant that can attack a wide range of targets. In this work, we studied the oxidation of hydrogen peroxide by peroxynitrite, which led to oxygen evolution. Oxygen yields increased at alkaline pH with an apparent pKa of 7.05 +/ 0.04. The maximum yields were 16% and 32% of added peroxynitrite at pH 5.9 and 7.4, respectively, assuming that two molecules of peroxynitrite are needed to produce one of oxygen. Hydroxyl radical scavengers (dimethyl sulfoxide, mannitol, ethanol, formate, and acetate) inhibited oxygen evolution to a similar extent to that predicted from their rate constants with hydroxyl radical. The apparent rate constant of peroxynitrite decomposition was zero-order in hydrogen peroxide at acidic pH. At neutral and alkaline pH, the rate of peroxynitrite disappearance decreased in the presence of millimolar concentrations of hydrogen peroxide by up to 50%. The apparent activation enthalpy and entropy for peroxynitrite decomposition increased by 1.7 kcal mol-1 and 4.7 cal mol-1 K-1, respectively, in the presence of hydrogen peroxide. We propose that an activated intermediate of peroxynitrous acid is responsible for hydrogen peroxide oxidation at acidic pH, while at more alkaline pH the formation of a stabilizing complex between hydrogen peroxide and transperoxynitrite anion is involved. PMID- 7492736 TI - Large scale synthesis of p-benzoquinone-2'-deoxycytidine and p-benzoquinone-2' deoxyadenosine adducts and their site-specific incorporation into DNA oligonucleotides. AB - Benzene is a carcinogen in rodents and a cause of bone marrow toxicity and leukemia in humans. p-Benzoquinone (p-BQ) is one of the stable metabolites of benzene, as well as of a number of drugs and other chemicals. 2'-Deoxycytidine (dC) and 2'-deoxyadenosine (dA) were allowed to react with p-BQ in aqueous solution at pH 7.4 and 4.5. The yields were considerably higher at pH 4.5 than at pH 7.4, as indicated by HPLC analysis. The desired products were isolated by column chromatography on silica gel or cellulose. Identification was done by FAB MS, 1H NMR, and UV spectroscopy. The reaction of p-BQ with dC and dA at pH 4.5 produced the exocyclic compounds 3-hydroxy-1,N4-benzetheno-2'-deoxycytidine (p-BQ dC), and 9-hydroxy-1,N6-benzetheno-2'-deoxyadenosine (p-BQ-dA), respectively, in a large scale and high yield. These adducts have been previously made in a microgram scale as the 3'-phosphate for 32P-postlabeling studies of their incidence in DNA. The p-BQ-dC and p-BQ-dA adducts have, in addition to the two hydroxyl groups of deoxyribose, one newly formed hydroxyl group at the C-3 or C-9 of the exocyclic base of each product respectively. Incorporation of these adducts into oligonucleotides as the phosphoramidite requires the protection of all three hydroxyl groups in these compounds. The exocyclic hydroxyl on the base has been successfully protected by acylation after protecting the 5'- and the 3' hydroxyl groups of the sugar moiety with a 4,4'-dimethoxytrityl group and a cyanoethyl N,N-diisopropylphosphoramidite group, respectively. For the first time, to our knowledge, the fully protected phosphoramidites of p-BQ-dC and p-BQ dA were prepared and incorporated site-specifically into a series of oligonucleotides. The coupling efficiency was very high (> 98%). However, deprotection of the DNA oligomers with ammonia produced only 50% of the desired oligomers containing the adduct. In contrast, when 10% of 1,8 diazabicyclo[5.4.0]undec-7-ene (DBU) in methanol at room temperature was used, only the desired oligomers were detected by HPLC. Thus, by deprotecting the oligomers with methoxide ions (DBU/methanol) and avoiding the use of ammonia, a high yield of modified DNA was obtained. After purification of these oligomers by HPLC, they were hydrolyzed enzymatically and analyzed by HPLC, which confirmed the base composition and the incorporation of the adducts. The mass spectroscopic analysis of the DNA oligomers was confirmed by electrospray MS. These oligomers are now under investigation for their biochemical properties. PMID- 7492737 TI - Prostaglandin H synthase-catalyzed ring oxygenation of 2-naphthylamine: evidence for two distinct oxidation pathways. AB - Previous studies showed that prostaglandin H synthase (PHS) cooxidizes 2 naphthylamine (2-NA) to ring-oxygenated products. These metabolites are atypical for a peroxidase-mediated reaction and are completely different from the polymeric nonoxygenated metabolites of 2-NA that are generated with the model peroxidase horseradish peroxidase (HRP). In this study, we investigated possible explanations for the PHS-catalyzed formation of ring-oxygenated 2-NA metabolites. We found that introduction of a peroxyl radical-generated cosubstrate into the HRP/2-NA system resulted in the formation of the same ring-oxygenated products observed in the PHS/2-NA system. 18O2 incorporation studies were utilized to further characterize the source of oxygen in the ring-oxygenated 2-NA metabolites. The data show that, in the case of PHS, ring oxygenation can occur both by peroxyl radical-mediated attack on 2-NA and by direct transfer of peroxide oxygen from PHS to 2-NA. PMID- 7492738 TI - The reductive conversion of chromium (VI) by ascorbate gives rise to apurinic/apyrimidinic sites in isolated DNA. AB - The formation of apurinic/apyrimidinic sites (AP-sites) and single strand breaks (SSB) by chromate and ascorbate (AsA) in isolated DNA was investigated using a number of agents that cleave DNA at AP-sites (putrescine, exonuclease III, the tripeptide Lys-Trp-Lys, and an AP-endonuclease containing fraction isolated from human fibroblasts). Relative to the number of SSB caused by chromate and AsA alone, all these agents induced additional nicking, indicating the induction of AP-sites. Chromate/AsA-induced AP-sites contain aldehyde groups, as cleavage by putrescine could be prevented by treatment with borohydride which reduced the aldehyde. The time course for the formation of both DNA lesions was very similar, and there was a 1:1 ratio of the number of SSB to the number of AP-sites. The addition of catalase to incubation mixtures containing chromate/AsA led to an almost complete suppression of AP-sites and SSB. In systems containing lower concentrations of chromate/AsA, the exclusion of oxygen inhibited the formation of both lesions. It is suggested that AP-sites and SSB arise from attack by reactive species deriving from chromate/AsA on one single site at DNA, probably the sugar moiety. In view of the known mutagenicity of AP-sites, these results could aid an understanding of the mechanisms underlying chromium(VI) carcinogenicity. PMID- 7492739 TI - Characterization of novel isocyanate-derived metabolites of the formamide N formylamphetamine with the combined use of electrospray mass spectrometry and stable isotope methodology. AB - Bioactivation of the formamide N-formylamphetamine (NFA) to 1-methyl-2 phenylethyl isocyanate (MPIC) was investigated in rats by screening bile and urine for conjugates subsequent to the phase I event. NFA was administered to rats as a mixture of protio- and pentadeuteriophenyl analogues to gain insight into the carbamoylating activity of MPIC when traced by electrospray liquid chromatography/mass spectrometry (LC/MS). An LC/MS contour generated by recording the summed mass spectrum as a function of chromatographic retention time allowed four biliary metabolites to be identified from four sets of doublets, with the peak of each doublet offset by 5 amu and ca. 0.07 min. Tandem mass spectrometry experiments allowed these metabolites to be attributed structurally to the glutathione, cysteinylglycine, cysteine, and N-acetylcysteine conjugates of the isocyanate MPIC. These assignments were subsequently validated by comparison of the LC/MS properties of the metabolites to synthetic reference compounds. Only the carbamoylated N-acetylcysteine conjugate was detected in urine. The observed excretion in bile of all metabolites of the mercapturate pathway is novel for formamide metabolism. NFA can thus be added to the short list of compounds that are eliminated in this fashion. Factors envisioned as contributory to this metabolic profile in bile include hepatorenal, enterohepatic, and biliary-hepatic cycling, in addition to possible equilibrium exchange of the isocyanate from thiocarbamate conjugates to endogenous free thiols during the course of biliary transit. PMID- 7492740 TI - Design and characterization of a model alpha beta peptide. AB - CD and nmr spectroscopy were used to compare the conformational properties of two related peptides. One of the peptides, Model AB, was designed to adopt a helix turn-extended strand (alpha beta) tertiary structure in water that might be stabilized by hydrophobic interactions between two leucine residues in the amino terminal segment and two methionine residues in the carboxyl terminal segment. The other peptide, AB Helix, has the same amino acid sequence as Model AB except that it lacks the -Pro-Met-Thr-Met-Thr-Gly segment at the carboxyl-terminus. Although the carboxyl-terminal segment of Model AB was found to be unstructured, its presence increases the number of residues in a helical conformation, shifts the pKas of three ionizable side chains by 1 pH unit or more compared to an unstructured peptide, stabilizes the peptide as a monomer in high concentrations of ammonium sulfate, increases the conformational stability of residues at the terminal ends of the helix, and results in many slowly exchanging amide protons throughout the entire backbone of the peptide. These results suggest that interactions between adjacent segments in a small peptide can have significant structure organizing effects. Similar kinds of interactions may be important in determining the structure of early intermediates in protein folding and may be useful in the de novo design of independently folding peptides. PMID- 7492741 TI - Energetics of protein-DNA interactions: an exact calculation for binding of ligands to a lattice of overlapping sites. AB - Exact equations are developed for analyzing the binding of ligands to a linear lattice of overlapping sites in which occupied-unoccupied as well as occupied occupied interactions are included for the analysis of the binding isotherms. We demonstrate that positive cooperativity on the binding of ligands to multiple sites may derive from either occupied-unoccupied or occupied-occupied interactions. When the binding of proteins to linear polynucleotides and DNA has exhibited positive cooperativity protein-protein (occupied-occupied), interactions have heretofore been invoked as the sole energetic source in determining the cooperative effect. Models and equations developed previously for the analysis of these binding isotherms have included only the protein-protein interactions (usually characterized with the symbol omega). The exact equations of this paper are capable of analyzing binding data in a manner to evaluate the relative importance of both occupied-unoccupied and occupied-occupied interactions. Relations derived here are employed to analyze some existing data, and the resulting parameter values are compared to those developed with equations employing only the protein-protein (occupied-occupied) interactions. The resulting parameter values are qualitatively different. Values of the binding constants differ by about three orders of magnitude. When only protein-protein interactions are taken into account, the resulting free energy of interaction is negative, indicating attractive forces between bound protein molecules; when both occupied-unoccupied and occupied-occupied interactions are applied, the resulting free energies of interaction are positive, indicating destabilizing forces acting primarily on the polynucleotide lattice. PMID- 7492742 TI - Multiple interconverting conformers of the cyclic tetrapeptide tentoxin, [cyclo (L-MeAla1-L-Leu2-MePhe[(Z) delta]3-Gly4)], as seen by two-dimensional 1H-nmr spectroscopy. AB - The conformations of the phytotoxic cyclic tetrapeptide tentoxin [cyclo-(L-MeAla1 L-Leu2-MePhe[(Z) delta]3-Gly4)] have been studied in aqueous solution by two dimensional proton nmr at various temperatures. Contrary to what is observed in chloroform, tentoxin exhibits multiple exchanging conformations in water. Aggregation phenomena were also observed. Four conformations with different proportions (51, 37, 8, and 4%) were observed at -5 degrees C. Models were constructed from nmr parameters and restrained molecular dynamics simulations. All the models exhibit cis-trans-cis-trans conformation of the amide bond sequence. The conversion from one form to another is accomplished by a conformational peptide flip consisting of a 180 degree rotation of a nonmethylated peptide bond. PMID- 7492743 TI - Characterization of the binding of YO to [poly(dA-dT)]2 and [poly(dG-dC)]2, and of the fluorescent properties of YO and YOYO complexed with the polynucleotides and double-stranded DNA. AB - The interaction between the fluorescent dye YO (oxazole yellow) and the alternating polynucleotides [poly(dA-dT)]2 [the duplex of alternating poly(dA dT)] and [poly(dG-dC)]2 [the duplex of alternating poly(dG-dC)] has been studied with optical spectroscopic techniques including absorbance, flow linear dichroism, CD, and fluorescence measurements. The principal features of the spectra are very similar for the two polynucleotide solutions, showing that YO binds quite similarly to AT and GC base pairs. From a strongly negative reduced linear dichroism (LDr) in the dye absorption band, an induced negative CD, and transfer of energy from the bases to bound YO, we conclude that at low mixing ratios YO is intercalated in both [poly(dA-dT)]2 and [poly(dG-dC)]2. At higher mixing ratios an external binding mode starts to contribute, evidenced from the appearance of an exciton CD. The conclusion that YO binds in a similar way to AT and GC base pairs should be valid also for the dimer YOYO since its YO units have been found to bind to double-stranded (dsDNA) in the same way as the YO monomer. The fluorescence properties of YO and YOYO complexed with DNA or the polynucleotides have been characterized by studying the dependence of fluorescence intensity on temperature, mixing ratio, and ionic strength. The fluorescence intensity and fluorescence lifetime of YO-DNA decrease strongly with increasing mixing ratio, whereas the fluorescence intensity of YOYO-DNA shows a weaker dependence, indicating that the quantum yield depends on the distance between the YO chromophores on the DNA chain. Further, the fluorescence intensity of YO depends on the base sequence; the quantum yield and fluorescence lifetime for YO complexed with [poly(dG-dC)]2 are about twice as large as for YO complexed with [poly(dA-dT)]2. Measurements of excitation spectra at different mixing ratios and different emission wavelengths indicate that the fluorescence of the externally bound chromophores is negligible compared to the intercalated ones. PMID- 7492744 TI - Conformational states governing the rates of spontaneous transition mutations. AB - We report the results of a theoretical study, combining the results of sequence analysis and integral equation structural methods for nucleic acids in aqueous solutions, on the effects of nearest neighbors on the (T.G) mispair in solution, for 12 nearest neighbor contexts. Attempts have been made to classify the structural and energetic effects of the 5' and 3' neighbors with respect to the observed spontaneous mutation rates in vertebrates. It is found that 5' nearest neighbor is probably the most critical structural factor in facilitating or discouraging mutations. Local conformational states correlate with discrimination of bases to be excised in mispairs. Our study confirms the role of the flexibility of the DNA molecule in governing the rates of spontaneous mutations. PMID- 7492746 TI - Docking of congo red to the surface of crystalline cellulose using molecular mechanics. AB - A method for the docking of ligands to crystalline surfaces using molecular mechanics was developed and applied to study the molecular features of the adsorption of Congo Red to cellulose crystals. The docking procedure was based on a grid search exploring the surface repeat unit of the cellulose crystals using several orientations of Congo Red at each grid point. Results suggest a preferential adsorption of Congo Red onto certain faces of the cellulose crystals and that the lower energy conformers have a similar positioning and orientation with respect to the cellulose chains at the surface repeat unit. PMID- 7492745 TI - Conformational mimicry: synthesis and solution conformation of a cyclic somatostatin hexapeptide containing a tetrazole cis amide bond surrogate. AB - Potent, cyclic hexapeptide analogues of somatostatin are generally believed to adopt some common secondary structural features: a II' beta turn at one end of the cycle, and a type VI turn with a cis amide bond at the other. A proposed cis amide surrogate, the 1,5-disubstituted tetrazole, has been placed into a cyclic hexapeptide analog of somatostatin in order to constrain the putative cis amide bond. The final cyclization was done by either chemical or enzymatic means. The product, cyclo(Ala6-Tyr7-D-Trp8-Lys9-Val10-Phe11-psi[CN4] ), was found to have 83% of the activity of somatostatin. Solution nmr analysis in DMSO/water revealed that the backbone as well as side chain chi1 and chi2 were well ordered. Relaxation matrix methods were used to extract distance restraints from the nuclear Overhauser effect spectroscopy data set, and these were used in a systematic search of torsional space to identify structures consistent with the nmr data. Restrained minimizations of these structures using a number of different force fields produced structures having the expected beta II' turn at D Trp8-Lys9 and a beta VIa turn in the Phe11-psi[CN4]-Ala6 portion of the molecule. The similarity of the minimized structures to those previously reported for cyclic hexapeptide analogues of somatostatin confirms the similarity of the tetrazole geometry to that of the cis amide in solution. PMID- 7492747 TI - Conformation of short DNA fragments by modulated fluorescence polarization anisotropy. AB - The technique of fluorescence polarization anisotropy (FPA) decay of intercalated ethidium has been used to study DNA conformation and dynamics, which are being recognized as primary determinants in transcription control and other cellular processes. Frequency modulated FPA when applied to two DNA molecules, a "straight" 50 base-pairs duplex fragment, and a bent fragment of similar length, has yielded different rotational diffusion coefficients for the two fragments. The data have been processed with an analytical model and with Brownian dynamics simulations, obtaining a good fit and a quantitative agreement between the two models. Both analyses have confirmed that one fragment can be described as a straight cylinder, while the other fragment is bent, with an angle estimated to be 45 degrees +/- 3 degrees. FPA has proved to be very powerful in determining simple conformations of short DNA duplexes and also particularly apt to probe the dynamical features of DNA fragments where conventional methods are either too cumbersome or fail to give quantitative results. In addition, the ligand no longer behaves ideally due to its complex structure and charge distribution. Thus for the protein the slope is no longer related simply to the net ligand charge, and the PB model gives a much larger slope than the LL model.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 7492748 TI - Salt effects on polyelectrolyte-ligand binding: comparison of Poisson-Boltzmann, and limiting law/counterion binding models. AB - The theory for salt dependence of the free energy, entropy, and enthalpy of a polyelectrolyte in the PB (PB) model is extended to treat the nonspecific salt dependence of polyelectrolyte-ligand binding reactions. The salt dependence of the binding constant (K) is given by the difference in osmotic pressure terms between the reactants and products. For simple 1-1 salts it is shown that this treatment is equivalent to the general preferential interaction model for the salt dependence of binding [C. Anderson and M. Record (1993) Journal of Physical Chemistry, Vol. 97, pp. 7116-7126]. The salt dependence, entropy, and enthalpy are compared for the PB model and one specific form of the preferential interaction coefficient model that uses counterion condensation/limiting law (LL) behavior. The PB and LL models are applied to three ligand-polyelectrolyte systems with the same net ligand charge: a model sphere-cylinder binding reaction, a drug-DNA binding reaction, and a protein-DNA binding reaction. For the small ligands both the PB and limiting law models give (In K vs. In[salt]) slopes close in magnitude to the net ligand charge. However, the enthalpy/entropy breakdown of the salt dependence is quite different. In the PB model there are considerable contributions from electrostatic enthalpy and dielectric (water reorientation) entropy, compared to the predominant ion cratic (release) entropy in the limiting law model. The relative contributions of these three terms in the PB model depends on the ligand: For the protein, ion release entropy is the smallest contribution to the salt dependence of binding. The effect of three approximations made in the LL model is examined: These approximations are (1) the ligand behaves ideally, (2) the preferential interaction coefficient of the polyelectrolyte is unchanged upon ligand binding, and (3) the polyelectrolyte preferential interaction coefficient is given by the limiting law/counterion condensation value. Analysis of the PB model shows that assumptions 2 and 3 break down at finite salt concentrations. For the small ligands the effects on the slope cancel, however, giving net slopes that are similar in the PB and LL models, but with a different entropy/enthalpy breakdown. For the protein ligand the errors from assumptions 2 and 3 in the LL models do not cancel. PMID- 7492749 TI - Comparison of three human-murine heteromyeloma cell lines for formation of human hybridomas after electrofusion with human peripheral blood lymphocytes from meningococcal cases and carriers. AB - Peripheral blood lymphocytes from meningitis patients and healthy meningococcal carriers were fused by electrofusion with the three human-murine heteromyeloma cell lines CB-F7, K6H6B5 and H7NS. 934 hybridomas producing human immunoglobulins were obtained in 30 fusions. Heteromyeloma K6H6B5 yielded a significantly higher proportion of hybridomas producing IgG antibodies than did the two other cell lines. CB-F7 and K6H6B5 yielded comparable numbers of hybridomas whose supernatants reacted with homologous bacteria, whereas the cell line H7NS was less efficient. PMID- 7492750 TI - Efficient immortalization of rheumatoid synovial tissue B-lymphocytes. A comparison between the techniques of electric field-induced and PEG fusion. AB - In this study, B-cells isolated from rheumatoid synovial tissue were immortalized, without prior in vitro stimulation, by means of electric-field induced fusion and conventional PEG fusion in order to compare the efficiency of these methods. Two myeloma cell lines were used as fusion partners, the murine myeloma Ag8 and the murine-human heteromyeloma HAB-1. The results of seven fusion experiments performed simultaneously with identical cell populations showed that fusion frequencies obtained by electrofusion were 4 to 35 times higher than by the PEG fusion technique. The morphological and immunohistochemical evaluation of synovial tissues used for fusion showed that only tissues exhibiting a follicular distribution of B-cells with a high percentage of CD 22-positive lymphocytes gave rise to high fusion yields and produced B-cell clones, whereas synovial tissues with the same percentage of plasma cells but lower percentages of CD 22 lymphocytes yielded very low fusion rates. In conclusion, electrofusion is more efficient for immortalizing small amounts of synovial tissue B-lymphocytes than PEG fusion, since high fusion frequencies could be obtained by this technique without the need for prior in vitro stimulation. Synovial tissue exhibiting a follicular distribution of B-lymphocytes with high percentages of CD 22-positive lymphocytes gave rise to high hybridoma yields and therefore an ideal source of human rheumatoid B-cell clones. PMID- 7492751 TI - Dual recognition of lipid A and DNA by human antibodies encoded by the VH4-21 gene: a possible link between infection and lupus. AB - The VH4-21 (V4-34) gene segment, a member of the VH4 family, is expressed early in B-cell maturation and is utilized by approximately 6% of normal adult B lymphocytes. This prevalence indicates an importance of VH4-21 in the B-cell repertoire. The gene also encodes certain autoantibodies being mandatory for pathological IgM anti-red cell antibodies directed against the I/i antigen, and also capable of encoding anti-DNA antibodies. Recognition of I/i antigen or DNA appears to be via two distinct sites on VH, with I/i binding mediated by sequences in the framework region, and DNA binding correlating with the presence of positively charged amino acids in complementarity-determining region 3. However, these positively charged residues appear to suppress the ability of the framework region to interact with I/i, rendering a single sequence monospecific for I/i or DNA. The IgM anti-DNA antibodies also recognize bacterial lipid A, whereas the anti-I/i antibodies do not, indicating that CDR3 may be involved in binding the negatively charged lipid A. Structural similarities between the DNA backbone and lipid A provide a possible explanation for this cross-reactivity. This dual recognition of bacterial antigen and autoantigen provides a potential link between infection and autoimmunity. PMID- 7492753 TI - Clinical evidence that the human monoclonal anti-idiotypic antibody 105AD7, delays tumor growth by stimulating anti-tumor T-cell responses. AB - A human monoclonal anti-idiotypic antibody, 105AD7, which mimics a colorectal tumor associated antigen, 791Tgp72, has been developed. A Phase I trial in advanced colorectal cancer patients showed that 105AD7 therapy was nontoxic and that immunised patients had prolonged survival when compared to a contemporary group of patients treated in the same center. There is accumulating clinical evidence that 105AD7 delays tumor growth by stimulating anti-tumor T-cell responses. Stimulation of helper T-cells was exemplified in the phase I study as 105AD7 immunized patients showed antigen specific T-cell blastogenesis responses and enhanced IL-2 production. Further evidence was obtained from a new clinical study in which colorectal cancer patients were immunized prior to tumor resection. Immune infiltrating cells were analysed by immunohistochemistry and effector cell function was studied in immune cells from peripheral blood and tumor draining lymph nodes. Both activated CD4 and natural killer (NK) cells were observed at the tumor site, which is of interest as NK cells are rarely found in colorectal tumors. Effector studies confirmed that NK activity was enhanced in 3/6 patients. Increased autologous tumor killing was also found in 3/4 patients and accumulation of CD8RO cells following 105AD7 immunization also suggested that CD8 T cells were being stimulated. PMID- 7492752 TI - A human-mouse chimeric Lym-1 monoclonal antibody with specificity for human lymphomas expressed in a baculovirus system. AB - A murine anti-human B-cell monoclonal antibody, Lym-1, has shown considerable promise for the treatment of human malignant lymphomas and has been utilized as a new radioimmunotherapy for refractory lymphoma. In order to enhance its clinical potential, a genetically engineered chimeric Lym-1 (chLym-1) with murine variable regions and human gamma 1 and kappa constant regions was constructed and expressed. The goal of this study was to generate a Lym-1 reagent with decreased immunogenicity and improved effector functions. Murine Lym-1 variable region cDNAs were isolated from the murine Lym-1 hybridoma cell line, fused to gamma 1 and kappa constant region cDNAs, and expressed in an insect cell expression system with the baculovirus transfer vector pAcUW31. The chLym-1 antibody expressed in this system was correctly processed and assembled into the expected immunoglobulin monomer. Chimeric Lym-1 bound to both target antigen-bearing Raji cells and a Lym-1 anti-idiotype antibody and had a similar binding affinity as murine Lym-1. The chimeric and murine versions of Lym-1 were assayed for their ability to mediate antibody-dependent cellular cytotoxicity (ADCC) and to induce complement-mediated cytotoxicity (CMC) against Raji targets. Chimeric Lym-1 mediated a two-fold higher level of ADCC than murine Lym-1 and slightly lower levels of CMC than murine Lym-1. In addition, in Raji lymphoma-bearing nude mice, chLym-1 localized to the tumor with approximately equal uptake at 24 and 72 hours. Chimeric Lym-1, however, cleared from the blood of nontumor-bearing mice approximately 5 times faster than murine Lym-1 (20 h vs. 5 days), as expected for a xenogeneic protein. The improved in vitro and in vivo activities of this genetically engineered monoclonal antibody render it a new potential immunotherapeutic reagent for the treatment of human malignant lymphomas. PMID- 7492754 TI - Comparison of anti-TNF alpha autoantibodies in plasma and from EBV transformed lymphocytes of autoimmune and normal individuals. AB - To examine the ability of normal and autoimmune individuals to produce circulating anti-TNF alpha antibodies, plasma samples from 10 RA patients, 10 SLE patients and 5 normal subjects were assessed for anti-TNF alpha antibody. While every individual tested demonstrated circulating IgM anti-TNF alpha antibody, IgG anti-TNF alpha autoantibody was seen predominantly in autoimmune patients. Only 1 of 5 normal individuals, but 15 of 20 autoimmune individuals had plasma IgG anti TNF alpha antibodies. To examine the ability of normal and autoimmune individuals to produce anti-TNF alpha autoantibody from their circulating lymphocytes, EBV transformation was performed. Oligoclonal immortal cell lines were successfully established from 13 patients and each one secreted detectable IgM anti-TNF alpha autoantibody. Transformed cells from only 1 of 5 normal individuals secreted IgM anti-TNF alpha autoantibody. These results indicate a higher prevalence of anti TNF alpha autoantibody production among autoimmune individuals although normal individuals are also capable of producing these autoantibodies. PMID- 7492755 TI - Generation of a human monoclonal antibody to hepatitis C virus, JRA1 by activation of peripheral blood lymphocytes and hypo-osmolar electrofusion. AB - We have generated a human monoclonal antibody with binding specificity for hepatitis C virus (HCV)-specific peptides using peripheral blood lymphocytes isolated from a HCV antibody positive patient. The B-lymphocytes were stimulated with lipopolysaccharide (LPS) for 72 hours prior to the fusion. A recently described high efficiency hypo-osmolar electrofusion technique was employed, allowing generation of a large number of human hybridomas. The hybridomas were screened for human immunoglobulin and HCV-specific peptide binding by EIA. A single HCV-positive clone, JRA1, was detected and sub-cloned. Isotype analysis showed it to secrete an IgM lambda monoclonal antibody. The antibody was positive on both first and second generation HCV antibody analysis. This study confirms that viable pathogen-specific B-cells may be recovered from the peripheral blood. Although such cells are likely to be relatively uncommon in the circulating B cell pool, they may be successfully immortalized by high efficiency electrofusion techniques. This technique might be valuable for the generation of human monoclonal antibodies with specificity for other human pathogens. PMID- 7492756 TI - Biological and clinical aspects of plasminogen activator inhibitor type 2. PMID- 7492757 TI - Chronic exposure to retroviral vector encoded MGDF (mpl-ligand) induces lineage specific growth and differentiation of megakaryocytes in mice. AB - Megakaryocyte growth and development factor (MGDF) has recently been identified as a ligand for the c-mpl receptor. Using retroviral-mediated gene transfer, MGDF has been overexpressed in mice to evaluate the systematic effects due to chronic exposure to this growth factor. MGDF overexpressing mice had more rapid platelet recovery than control mice after transplantation. Following this recovery, the platelet levels continued increasing to fourfold to eightfold above normal baseline levels and remained elevated (five-fold above control mice) in these animals, which are alive and well at more than 4 months posttransplantation. Increased megakaryocyte numbers were detected in a number of organs in these mice including bone marrow, spleen, liver, and lymph nodes. Prolonged overexpression of MGDF led to decreased marrow hematopoiesis, especially erythropoiesis, with a shift to extramedullary hematopoiesis in the spleen and liver. All the MGDF overexpressing mice analyzed to date developed myelofibrosis and osteosclerosis, possibly induced by megakaryocyte and platelet produced cytokines. No significant effect on other hematopoietic lineages was seen in the MGDF overexpressing mice, showing that the stimulatory effect of MGDF in vivo is restricted to the megakaryocyte lineage. PMID- 7492758 TI - Carrier frequency of the IVS4 + 4 A-->T mutation of the Fanconi anemia gene FAC in the Ashkenazi Jewish population. AB - Fanconi anemia (FA) is a genetically and phenotypically heterogeneous autosomal recessive disorder defined by a cellular hypersensitivity to DNA cross-linking agents. One of the FA genes, FAC, has been cloned and the genomic structure of the coding region has been characterized. We have developed amplification refractory mutation system (ARMS) assays for five known mutations in FAC, and have applied these assays to determine the carrier frequency of the IVS4 + 4 A- >T (IVS4) mutation in an Ashkenazi Jewish population. We tested 3,104 Jewish individuals, primarily of Ashkenazi descent, for the two most common FAC mutations, IVS4 and 322delG. Thirty-five IVS4 carriers were identified, for a carrier frequency of 1 in 89 (1.1%; 95% confidence interval 0.79% to 1.56%); no 322delG carriers were found. To determine if the IVS4 mutation was confined to the Ashkenazi Jewish population, we tested 563 Iraqi Jews for IVS4, and no carriers were found. Because the IVS4 mutation has only been found on chromosomes of Ashkenazi Jewish origin and is the only FAC mutation found on these chromosomes, we suggest that a founder effect is responsible for the high frequency of this mutation. With a carrier frequency greater than 1% and simple testing available, the IVS4 mutation merits inclusion in the battery of tests routinely provided to the Jewish population. PMID- 7492759 TI - IgE-independent interleukin-4 expression and induction of a late phase of leukotriene C4 formation in human blood basophils. AB - T-helper cells can differentiate into at least two subtypes secreting distinct profiles of cytokines, Th1 and Th2, regulating immunoprotection and different immunopathologies. Interleukin-4 (IL-4) is both the product and the inducer of Th2 cells, raising the question whether IL-4 can be produced in response to antigen-independent stimuli. Here we show that human basophils produce IL-4 on stimulation with IL-3 and C5a or C5adesarg in similar amounts as induced by IgE receptor-cross-linking. C5a-induced IL-4 production requires the presence of IL 3, with little effect of the sequence of stimuli addition. No "Th1-cytokines" (interferon-gamma and IL-2) and even no "Th2-cytokines" (IL-3, IL-5, IL-10, and granulocyte-macrophage colony-stimulating factor) are produced by basophils in response to either IgE-dependent or IgE-independent activation. The generation of leukotriene C4 (LTC4) is regulated in a similar manner. However, C5a induces a rapid, transient burst of leukotriene formation only if added after IL-3. Interestingly, upon prolonged culture, a late phase of continuous LTC4 production is observed, which also requires two signals (IL-3 and C5a), but rather depends on their continuous presence than on their sequence of action. These data describe an antigen-independent pathway of very restricted IL-4 expression. Thus, basophils must be considered as central immunoregulatory cells of the innate immune system. Furthermore, the results show that LTC4 can also be generated more continuously for many hours, a phenomenon that may be of particular importance in chornic allergic inflammation, such as asthma. PMID- 7492760 TI - Molecular basis for the recently described hereditary hyperferritinemia-cataract syndrome: a mutation in the iron-responsive element of ferritin L-subunit gene (the "Verona mutation") AB - Recently, we described a new genetic disorder (the "hereditary hyperferritinemia cataract syndrome") clinically characterized by the combination of elevated serum ferritin and congenital bilateral nuclear cataract, both cotransmitted as an autosomal dominant trait. In affected subjects, hyperferritinemia (ranging from 950 to 2,259 micrograms/L) is typically not related to iron overload. Differently from subjects with hereditary hemochromatosis, they have normal to low levels of serum iron and percent of transferrin saturation and absence of iron overload in parenchymal organs. When unnecessary phlebotomies are performed, they rapidly develop iron-deficient anemia, with persistently elevated levels of serum ferritin. By RNA-single-strand conformation polymorphism screening of the L subunit ferritin gene on chromosome 19, we were able to identify in affected subjects a mutation in the 5' untranslated region. This mutation involves the five nucleotides sequence [CAGUG] of the iron-responsive element (IRE), which is critical for the posttranscriptional regulation of ferritin synthesis by means of IRE-binding protein (IRE-BP). Thus, it is very likely to provide the molecular basis for the iron-insensitive upregulation of ferritin synthesis in affected subjects. PMID- 7492761 TI - Regulation of platelet activation in vitro by the c-Mpl ligand, thrombopoietin. AB - Thrombopoietin (TPO) is a recently identified growth factor that regulates megakaryocytopoiesis. Its receptor, c-Mpl, is expressed in megakaryocyte progenitors, mature megakaryocytes, and human blood platelets. We have observed that TPO treatment of human platelets resulted in tyrosine phosphorylation of several cellular proteins, including the c-Mpl receptor and the 85-kD subunit of phosphatidylinositol 3-kinase (PI3-K). TPO stimulated this tyrosine phosphorylation in a time-dependent manner, reaching a maximum in 5 minutes. The tyrosine phosphorylation of PI 3-K was dependent on the concentration of TPO and reached a maximum at concentrations between 50 and 100 ng/mL. This phosphorylation was independent of extracellular fibrinogen and ligation of the alpha IIb beta 3 integrin. In contrast, TPO, in the presence of exogenous fibrinogen, induced concentration-dependent platelet aggregation, which was blocked by the soluble c-Mpl receptor. Increasing TPO concentrations modulated the degree of the primary wave of aggregation and the lag phase, but not the slope or maximum of the secondary wave of aggregation. This secondary aggregation was controlled by the addition of apyrase, suggesting an adenosine diphosphate (ADP)-dependent mechanism. Treatment of platelets with TPO resulted in augmented binding of 125I-fibrinogen to intact platelets, with a 50% effect (EC50) occurring between 5 and 10 ng/mL. TPO-induced binding of fibrinogen to platelets was comparable in degree with that observed by stimulation with 10 mumol/L ADP. In an immobilized collagen-platelet adhesion assay, a significant increase in the attachment of TPO-stimulated platelets was observed. This effect was dependent on the concentration of TPO. At 50 ng/mL of TPO, platelet attachment to collagen increased threefold compared with the buffer control. Furthermore, the presence of fibrinogen did not significantly alter TPO augmentation of the platelet collagen interaction. This interaction was mediated by the Arg-Gly-Asp (RGD) adhesion recognition sequence, as it was completely abolished by 100 mumol/L of the RGDS peptide. A fraction of the TPO-dependent platelet attachment to a collagen-coated surface was insensitive to treatment with prostaglandin E1. Furthermore, antibody to alpha IIb integrin partially inhibited platelet attachment to collagen, suggesting that the integrin alpha IIb beta 3 participates in this association. These data indicate that TPO might function not only as a cytokine in megakaryocyte growth and differentiation, but may also participate in direct platelet activation and modulate platelet-extracellular matrix interactions. PMID- 7492762 TI - Radioimmunotherapy of interleukin-2R alpha-expressing adult T-cell leukemia with Yttrium-90-labeled anti-Tac. AB - Adult T-cell leukemia (ATL) is a malignancy of mature lymphocytes caused by the retrovirus human T-cell lymphotropic virus-I. It is an aggressive leukemia with a median survival time of 9 months; no chemotherapy regimen appears successful in inducing long-term disease-free survival. The scientific basis of the present study is that ATL cells express high-affinity interleukin-2 receptors identified by the anti-Tac monoclonal antibody, whereas normal resting cells do not. To exploit this difference, we administered anti-Tac armed with Yttrium-90 (90Y) to 18 patients with ATL initially (first 9 patients) in a phase I dose-escalation trial and subsequently (second group of 9 patients) in a phase II trial involving a uniform 10-mCi dose of 90Y-labeled anti-Tac. Patients undergoing a remission were permitted to receive up to eight additional doses. At the 5- to 15-mCi doses used, 9 of 16 evaluable patients responded to 90Y anti-Tac with a partial (7 patients) or complete (2 patients) remission. The responses observed represent improved efficacy in terms of length of remission when compared with previous results with unmodified anti-Tac. Clinically meaningful (> or = grade 3) toxicity was largely limited to the hematopoietic system. In conclusion, radioimmunotherapy with 90Y anti-Tac directed toward the IL-2R expressed on ATL cells may provide a useful approach for treatment of this aggressive malignancy. PMID- 7492763 TI - Separation of myeloid and erythroid progenitors based on expression of CD34 and c kit. AB - In this report, a novel approach is described to physically separate erythroid progenitors from monocyte and granulocyte progenitors, based on the expression of CD34 and Kit. Using biotin-labeled human Kit ligand (KL) and flow cytometry, Kit was detectable on 2% to 3% of the nucleated cells in rhesus monkey bone marrow. Combination of biotin-KL with CD34 monoclonal antibodies (MoAb) showed that Kit was expressed on subsets of CD34low and CD34pos cells. Our data clearly demonstrate that CD34pos cells are more heterogeneous with respect to Kit expression than observed in studies using Kit MoAb. A small cluster, approximately 7% of the CD34pos cells, expressed CD34 at submaximal levels and stained brightly with biotinylated KL. This CD34pos/kithi fraction contained predominantly erythroid progenitors (burst-forming units-erythroid; BFU-E). The majority of the granulocytic and monocytic progenitors (colony-forming units granulocyte/macrophage; CFU-GM) were CD34pos/kitmed. Some BFU-E were also detected in the CD34pos/kitmed and CD34low/kitpos fractions at low frequency. In the latter subset, most erythroid colony-forming units (CFU-E) were recovered. Using three-color flow cytometry, we analyzed expression of Kit in relation to that of CD34 and the class II major histocompatibility antigen, RhLA-DR. The most immature bone marrow cells that can be identified in vitro, ie, CD34pos/RhLA DRlow cells, were kitmed. The CD34pos/kithi and CD34pos/kitneg subsets predominantly contained the more mature RhLA-DRbright cells. Our results demonstrate that erythroid precursors express c-kit at much higher levels than monomyeloid precursors and pluripotent progenitors. The difference in expression levels of CD34 and c-kit can be exploited to isolate BFU-E populations that are virtually devoid of nonerythroid cells. PMID- 7492764 TI - Detection of cytomegalovirus DNA in CD34+ cells from blood and bone marrow. AB - Infection of hematopoietic progenitor cells with the human cytomegalovirus (HCMV) has been proposed as an explanation for the cytopenias associated with HCMV related disease. To test this hypothesis, CD34+ cells, which include the hematopoietic progenitors, as well as mature leukocyte populations were purified on a fluorescence-activated cell sorter and analyzed for HCMV DNA by polymerase chain reaction (PCR). A total of 33 samples from 31 immunosuppressed as well as immunocompetent HCMV-seropositive individuals were studied. CD34+ cells were PCR positive in four of seven bone marrow aspirates from allogeneic bone marrow transplant recipients, in three of eight aspirates from patients with acquired immunodeficiency syndrome, and in the first of two bone marrow samples from an immunocompetent patient with primary HCMV disease. CD34+ cells purified from peripheral blood for autologous and allogeneic transplantation were also analyzed, and 4 of 13 samples were HCMV DNA-positive. Interestingly, two of the four HCMV-positive samples were from healthy allogeneic donors. Among the mature leukocyte populations, the monocytes were most frequently found to be HCMV DNA positive. No HCMV DNA was detected in the total bone marrow leukocytes of 13 healthy seropositive bone marrow donors or in the CD34+ cell fraction of three further seropositive donors. In conclusion, the data provide strong evidence that CD34+ hematopoietic progenitor cells can be infected with HCMV in immunosuppressed patients, while this cell population was not identified as a major viral reservoir in healthy HCMV-seropositive individuals. PMID- 7492765 TI - Plasma/serum levels of flt3 ligand are low in normal individuals and highly elevated in patients with Fanconi anemia and acquired aplastic anemia. AB - The flt3 ligand is a growth factor that stimulates the proliferation of hematopoietic progenitor and stem cells. We established a sensitive enzyme-linked immunosorbent assay (ELISA) to measure the concentration of flt3 ligand in plasma or serum from normal individuals, as well as in patients with hematopoietic disorders. Concentrations of flt3 ligand in plasma or serum from normal individuals were quite low: only 12% (7 of 60) of normal individuals had flt3 ligand levels above 100 pg/mL (the limit of detection). In contrast, 86% (19 of 22) of samples from patients with Fanconi anemia and 100% (eight of eight) of samples from patients with acquired aplastic anemia had plasma or serum levels above 100 pg/mL. Mean plasma or serum concentrations (calculated by assigning a value of 0 pg/mL to any sample reading below the level of detection) were as follows: normal volunteers, 14 pg/mL; patients with Fanconi anemia, 1,331 pg/mL; and patients with acquired aplastic anemia, 460 pg/mL. Concentrations of flt3 ligand in blood are, therefore, specifically elevated to a level that may be physiologically relevant in hematopoietic disorders with a suspected stem cell component. The elevated flt3 ligand concentrations in these individuals may be part of a compensatory hematopoietic response to boost the level of progenitor cells. PMID- 7492766 TI - Role of antennary structure of N-linked sugar chains in renal handling of recombinant human erythropoietin. AB - To elucidate the role of the branched structure of sugar chains of human erythropoietin (EPO) in the expression of in vivo activity, the pharmacokinetic profile of a less active recombinant human EPO sample (EPO-bi) enriched with biantennary sugar chains was compared with that of a highly active control EPO sample enriched with tetraantennary sugar chains. After an intravenous injection in rats, 125I-EPO-bi disappeared from the plasma with 3.2 times greater total body clearance (Cltot) than control 125I-EPO. Whole-body autoradiography after 20 minutes of administration indicated that the overall distribution of radioactivity is similar, but 125I-EPO-bi showed a higher level of radioactivity in the kidneys than control 125I-EPO. Quantitative determination of radioactivity in the tissues also indicated that radioactivity of 125I-EPO-bi in the kidneys was two times higher than that of control 125I-EPO. The difference in plasma disappearance between 125I-EPO-bi and control 125I-EPO was not observed in bilaterally nephrectomized rats. The distribution of 125I-EPO-bi to bone marrow and spleen was similarly inhibited by simultaneous injection of excess amounts of either the nonlabeled EPO-bi or control EPO. These results indicate that the low in vivo biologic activity of EPO-bi results from rapid clearance from the systemic circulation by renal handling. Thus, the well-branched structure of the N-linked sugar chain of EPO is suggested to play an important role in maintaining its higher plasma level, which guarantees an effective transfer to target organs and stimulation of erythroid progenitor cells. PMID- 7492767 TI - Effects of human FLT3 ligand on myeloid leukemia cell growth: heterogeneity in response and synergy with other hematopoietic growth factors. AB - A novel hematopoietic growth factor for primitive hematopoietic progenitor cells, the ligand for the flt3/flk2 receptor, (FL), has been recently purified and its gene has been cloned. In the present study, we investigated the effects of FL on the proliferation and differentiation of normal and leukemic myeloid progenitor cells. We demonstrate that FL is a potent stimulator of the in vitro growth of granulocyte-macrophage colony-stimulating factor (GM-CSF), interleukin-3 (IL-3), or G-CSF-dependent granulocyte-macrophage committed precursors from Lin- CD34+ bone marrow cells of normal donors. By contrast, FL does not affect the growth of erythroid-committed progenitors even in the presence of erythropoietin. The effect of FL on the proliferation and on the in vitro growth of clonogenic leukemic precursor cells was studied in 54 acute myeloid leukemia (AML) cases. Fresh leukemia blasts from 36 of 45 patients with AML significantly responded to FL without any relation to the French-American-British (FAB) subtype. FL stimulated the proliferation of leukemic blasts in a dose-dependent fashion. Synergistic activities were seen when FL was combined with G-CSF, GM-CSF, IL-3, or stem cell factor (SCF). FL as a single factor induced or increased significantly colony formation by clonogenic precursor cells from 21 of 24 patients with AML. In the presence of suboptimal and optimal concentrations of G CSF, GM-CSF, IL3, SCF, or a combination of all factors, FL strongly enhanced the number of leukemic colonies (up to 18-fold). We also evaluated the induction of tyrosine phosphorylated protein on FL stimulation in fresh AML cells. We demonstrate that, on FL stimulation, a band of phosphorylated protein(s) of about 90 kD can be detected in FL-responsive, but not in FL-unresponsive cases. This study suggests that FL may be an important factor for the growth of myeloid leukemia cells, either as a direct stimulus or as a synergistic factor with other cytokines. PMID- 7492768 TI - Granulomonocyte-associated lysosomal protein expression during in vitro expansion and differentiation of CD34+ hematopoietic progenitor cells. AB - Using an in vitro expansion and differentiation system for human CD34+ cord blood (CB) progenitor cells, we analyzed the induction and expression kinetics of the granulomonocyte associated lysosomal proteins myeloperoxidase (MPO), lysozyme (LZ), lactoferrin (LF), and macrosialin (CD68). Freshly isolated CD34+ CB cells were negative for LZ and LF, and only small proportions expressed MPO (4% +/- 2%) or CD68 (3% +/- 1%). Culturing of CD34+ cells for 14 days with interleukin (IL) 1, IL-3, IL-6, stem cell factor, granulocyte-macrophage colony-stimulating factor (GM-CSF), and G-CSF resulted in on average a 1,750-fold amplification of cell number, of which 83% +/- 7% were MPO+. Without addition of GM-CSF and G-CSF, lower increases in total cell numbers (mean, 211-fold) and lower proportions of MPO+ cells (54% +/- 11%) were observed. The proportion of MPO+ cells slightly exceeded but clearly correlated with the proportion of cells positive for the granulomonocyte associated surface molecules CD11b (Mac-1), CD15 (LeX), CD64 (Fc gamma RI) CD66, or CD89 (Fc alpha R). At day 14 MPO+ and LZ+ cells were virtually identical. However, at earlier time points during culture (days 4 and 7), single MPO+ or LZ+ cell populations were also observed, which only later acquired LZ and MPO, respectively. Maturation of cells into the neutrophilic pathway was indicated by the acquisition of MPO, followed by LZ. In contrast, maturation of cells into the monocytic pathway was indicated by the acquisition of LZ followed by MPO and CD14. CD68 was found to be expressed at day 4 by the majority of cells and was not restricted to the granulomonocytic cells, as cells with megakariocytic (CD41+) or erythroid (CD71hi) features were CD68+. LF expression was observed only in GM- plus G-CSF-supplemented cultures, in which only 26% +/- 5% of cells expressed LF by day 14. PMID- 7492771 TI - Salicylates inhibit lipopolysaccharide-induced transcriptional activation of the tissue factor gene in human monocytic cells. AB - Binding of plasma Factor VII/VIIa to the tissue factor (TF) receptor initiates the coagulation protease cascades. TF expression by circulating monocytes is associated with thrombotic and inflammatory complications in a variety of diseases. Transcriptional activation of the human TF gene in monocytic cells exposed to bacterial lipopolysaccharide (LPS) is mediated by binding of c-Rel/p65 heterodimers to a kappa B site in the TF promoter. Here, we report that a family of anti-inflammatory agents, known as the salicylates, inhibited LPS induction of TF activity and TF gene transcription in human monocytes and monocytic THP-1 cells at clinically relevant doses. Furthermore, sodium salicylate blocked the LPS-induced proteolytic degradation of I kappa B alpha, which prevented the nuclear translocation of c-Rel/p65 heterodimers. In contrast, two other nonsteroidal anti-inflammatory drugs, ibuprofen and indomethacin, did not inhibit LPS induction of the TF gene. These results indicated that salicylates inhibited LPS induction of TF gene transcription in monocytic cells by preventing nuclear translocation of c-Rel/p65 heterodimers. The clinical benefits of salicylates in the treatment of several diseases, including atherosclerosis and rheumatoid arthritis, may be related to their ability to reduce monocyte gene expression. PMID- 7492772 TI - Procoagulant activity of reversibly acylated human factor Xa. AB - The plasma clotting factors used to treat hemophiliacs who have developed inhibitory antibodies have a shared history of limited clinical safety and utility. To improve on existing bypass factors, we have developed a reversibly acylated form of human plasma factor Xa capable of providing a time-dependent release of procoagulant activity. Factor Xa was treated with p-amidinophenyl p' anisate to generate anisoyl Xa. The chemical modification of the protein involves acylation of the active site serine residue of factor Xa. Anisoyl Xa deacylated in a time, pH, and temperature-dependent manner. Active factor Xa generated on deacylation of anisoyl Xa exhibited amidolytic and prothrombinase complex activities in in vitro assays, the level being comparable to those of untreated factor Xa. When Anisoyl Xa was infused into rabbits, active factor Xa was generated on deacylation of the acylated enzyme, which shortened the activated partial thromboplastin time (APTT) in a dose-dependent manner. The duration of effect on rabbit APTT could be directly correlated to the level of human plasma factor Xa. Because anisoyl Xa bypasses the "tenase" complex that is compromised in hemophilia A and B and is unaffected by inhibitory antibodies, it has the potential to be used as an effective bypass therapy. PMID- 7492770 TI - Monoclonal antibody F1 binds to the kringle domain of factor XII and induces enhanced susceptibility for cleavage by kallikrein. AB - In a previous study we have shown that monoclonal antibody F1 (MoAb F1), directed against an epitope on the heavy chain of factor XII distinct from the binding site for anionic surfaces, is able to activate factor XII in plasma (Nuijens JH, et al: J Biol Chem 264; 12941, 1989). Here, we studied in detail the mechanism underlying the activation of factor XII by MoAb F1 using purified proteins. Formation of factor XIIa was assessed by measuring its amidolytic activity towards the chromogenic substrate H-D-Pro-Phe-Arg-pNA (S-2302) in the presence of soybean trypsin inhibitor and by assessing cleavage on sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE). Upon incubation with MoAb F1 alone, factor XII was auto-activated in a time-dependent fashion, activation being maximal after 30 hours. Factor XII incubated in the absence of MoAb F1 was hardly activated by kallikrein, whereas in the presence of MoAb F1, but not in that of a control MoAb, the rate of factor XII activation by kallikrein was promoted at least 60-fold. Maximal activation of factor XII with kallikrein in the presence of MoAb F1 was reached within 1 hour. This effect of kallikrein on the cleavage of factor XII bound to MoAb F1 was specific because the fibrinolytic enzymes plasmin, urokinase, and tissue-type plasminogen activator could not substitute for kallikrein. Also, trypsin could easily activate factor XII, but in contrast to kallikrein, this activation was independent of MoAb F1. SDS-PAGE analysis showed that the appearance of amidolytic activity correlated well with cleavage of factor XII. MoAb F1-induced activation of factor XII in this purified system was not dependent on the presence of high-molecular-weight kininogen (HK), in contrast to the activation of the contact system in plasma by MoAb F1. Experiments with deletion mutants revealed that the epitopic region for MoAb F1 on factor XII is located on the kringle domain. Thus, this study shows that binding of ligands to the kringle domain, which does not contribute to the proposed binding site for negatively charged surfaces, may induce activation of factor XII. Therefore, these findings point to the existence of multiple mechanisms of activation of factor XII. PMID- 7492769 TI - Dysregulated expression of GATA-1 following retrovirus-mediated gene transfer into murine hematopoietic stem cells increases erythropoiesis. AB - Retrovirus-mediated gene transfer was used to study the effects of dysregulated expression of the zinc-finger transcription factor, GATA-1, which has been shown to be required for erythropoiesis. A retroviral vector (PGK-GATA-1) was constructed with the murine GATA-1 gene linked to the human phosphoglycerate kinase (PGK) promoter. Expression of GATA-1 was demonstrated by super-shift analysis with a monoclonal antibody against murine GATA-1 using extracts of nonerythroid cytotoxic T-lymphocyte line (CTLL) cells transduced with the PGK GATA-1 virus. Mouse bone marrow cells were transduced in vitro and transplanted into recipient animals. Polymerase chain reaction (PCR) analysis performed on DNA extracted from peripheral blood 12 to 40 weeks posttransplantation demonstrated the presence of the PGK-GATA-1 provirus. Proviral integrity and copy number were demonstrated by Southern blot analysis of DNA from spleen, thymus, and bone marrow tissues from the long-term animals. At 16 weeks posttransplant, animals that received cells transduced by the GATA-1 virus maintained a lower white blood cell (WBC) count and absolute neutrophil count (ANC) and a higher red blood cell (RBC) count than control animals that received cells transduced with a virus containing a neor gene. Erythropoiesis was stimulated in GATA-1 and control animals by phlebotomy. GATA-1 animals required more extensive phlebotomy to reach a hematocrit less than 25 and their hematocrit returned to normal levels sooner than control animals. The effect of twice-daily injections of 10 U recombinant erythropoietin (epo) was also examined. The hematocrit of GATA-1 animals showed a more rapid and elevated response to epo than the hematocrit of control animals. These data suggest that dysregulated expression of GATA-1 in primitive hematopoietic cells enlarges the pool of epo-responsive erythroid progenitor cells. PMID- 7492773 TI - On the role of von Willebrand factor in promoting platelet adhesion to fibrin in flowing blood. AB - Platelet adhesion to fibrin at high shear rates depends on both the glycoprotein (GP) IIb:IIIa complex and a secondary interaction between GPIb and von Willebrand factor (vWF). This alternative link between platelets and vWF in promoting platelet adhesion to fibrin has been examined in flowing whole blood with a rectangular perfusion chamber. Optimal adhesion required both platelets and vWF, as shown by the following observations. No binding of vWF could be detected when plasma was perfused over a fibrin surface or when coated fibrinogen was incubated with control plasma in an enzyme-linked immunosorbent assay. However, when platelets were present during perfusion, interactions between vWF and fibrin could be visualized with immunoelectron microscopy. Exposure of fibrin surfaces to normal plasma before perfusion with severe von Willebrand's disease blood did not compensate for the presence of plasma vWF necessary for adhesion. vWF mutants in which the GPIIb:IIIa binding site was mutated or the GPIb binding site was deleted showed that vWF only interacts with GPIb on platelets in supporting adhesion to fibrin and not with GPIIb:IIIa. Complementary results were obtained with specific monoclonal antibodies against vWF. Thus, vWF must first bind to platelets before it can interact with fibrin and promote platelet adhesion. Furthermore, only GPIb, but not GPIIb:IIIa is directly involved in this interaction of vWF with platelets. PMID- 7492774 TI - An essential role for lysophosphatidylcholine in the inhibition of platelet aggregation by secretory phospholipase A2. AB - The release of secretory phospholipase A2 (sPLA2) into the mammalian circulation may contribute to the development of hemorrhagic and inflammatory diseases. sPLA2 has previously been shown to alter the behavior of platelets, leukocytes, and endothelial cells, although the molecular basis for these cellular effects has not been established. Our studies indicate that the inhibition of platelet aggregation by snake, bee venom, and pancreatic sPLA2 is dependent on a plasma cofactor. This cofactor resides within the lipoprotein fraction of plasma, with 54%, 31%, and 11% of the activity present in the high-density lipoprotein (HDL), low-density lipoprotein (LDL), and very low density lipoprotein (VLDL) fractions, respectively. Delipidation of HDL and LDL was associated with the complete loss of platelet-inhibitory activity. Incubation of purified sPLA2 with the HDL fraction of plasma resulted in the time-dependent generation of lysophosphatidylcholine (lysoPC). The formation of lysoPC correlated with the inhibition of platelet aggregation. Purified lysoPC (10 to 100 micrograms/mL) inhibited platelet aggregation and dense granule release induced by thrombin (0.05 U/mL), collagen (1 micrograms/mL), ionophore A23187 (2 mumol/L), ADP (12.5 mumol/L), and adrenaline (3.2 mumol/L). The inhibition of platelet aggregation by lysoPC was dose-dependent and correlated with decreased fibrinogen binding to glycoprotein IIb-IIIa. Our studies indicate that the enzymatic generation of lysoPC from plasma lipoproteins is essential for the sPLA2-mediated inhibition of platelet activation in the presence of albumin. These results raise the possibility that the toxic effects of circulating sPLA2 may be due in part to the generation of the bioactive lysophospholipid, lysoPC. PMID- 7492775 TI - Aggregation of mammalian cells expressing the platelet glycoprotein (GP) Ib-IX complex and the requirement for tyrosine sulfation of GP Ib alpha. AB - The glycoprotein (GP) Ib-IX complex mediates platelet aggregation in response to high shear forces by binding von Willebrand factor (vWF) in the plasma. We investigated the possibility that the complex could mediate a similar phenomenon if expressed in nonhematopoietic cells. When agitated on a tabletop shaker, CHO and L cells expressing the full complex formed large aggregates in the presence of vWF and the modulator ristocetin. When the rate of agitation was increased, aggregation occurred without added ristocetin and appeared to require only the application of a physical force. The aggregation was homophilic and temperature dependent and required a functional ligand-binding subunit of the GP Ib-IX complex, GP Ib alpha. Posttranslational tyrosine sulfation of GP Ib alpha was required for aggregate formation and stability. Thus, aggregation of cells expressing the GP Ib-IX complex is a unique example of a ligand-receptor interaction induced by mechanical forces and demonstrates an important biological role for sulfation of tyrosine residues. PMID- 7492776 TI - Critical involvement of transmembrane tumor necrosis factor-alpha in endothelial programmed cell death mediated by ionizing radiation and bacterial endotoxin. AB - In this report, we show that ionizing radiation (IR) at a clinically relevant dose (4 Gy) causes apoptosis in macrovascular and microvascular human endothelial cells. Treatment of irradiated cells with a low dose of bacterial endotoxin (LPS), similar to the levels observed in serum during endotoxemia, enhanced the rate of apoptosis, although LPS alone was unable to induce programmed cell death. The cytokine and endotoxin antagonist interleukin-10 (IL-10) reduced the rate of LPS + IR-induced apoptosis to levels obtained with irradiation alone. Using neutralizing antibodies against tumor necrosis factor-alpha (TNF), we could show crucial involvement of TNF in the LPS-mediated enhancement of IR-induced apoptosis, but not in the IR-induced apoptosis per se. However, further analysis strongly suggested the transmembrane form of TNF (mTNF), but not soluble TNF, to be accountable for the LPS-mediated cytotoxic effects. Studies with anatagonistic receptor specific antibodies clearly showed that TNF receptor type I (TR60) is essential and sufficient to elicit this effect. These findings are of potential clinical importance because they may disclose a relevant mechanism that leads to endothelial damage after radiotherapy or total body irradiation used for conditioning in bone marrow transplantation and that may thus contribute to transplant related complications, especially in association with endotoxemia or related inflammatory states. PMID- 7492777 TI - CD43 regulates tyrosine phosphorylation of a 93-kD protein in T lymphocytes. AB - The leukocyte sialyloglycoprotein CD43 exhibits features of a signal transducing molecule and is thought to be important for T-cell activation and adhesion. However, cellular biochemical events in which CD43 participates remain poorly understood. Here we provide evidence that CD43 regulates tyrosine phosphorylation of a specific substrate in T cells. A 93-kD tyrosine phosphoprotein was identified specifically in the CD43+ T-cell line CEM, but not in their CD43 deficient counterparts derived by gene targeting. The 93-kD phosphoprotein was detected in the CD43-deficient CEM cells after transfection with CD43 cDNA, and it could be specifically phosphorylated in lysates from the CD43-deficient cells by incubation with a CD43 immunoprecipitate obtained from the CD43+ cells. Expression of CD43 in HeLa cell transfectants was associated with the appearance of novel phosphoproteins including one with a molecular weight of approximately 93 kD, confirming that tyrosine phosphorylation of cellular substrates results specifically from CD43 expression. We conclude that CD43 regulates tyrosine phosphorylation of a 93-kD T-cell substrate. PMID- 7492778 TI - Spontaneous and glucocorticoid-induced apoptosis in human mature T lymphocytes. AB - Glucocorticoid (GC)-induced apoptosis is a well-recognized physiologic regulator of murine T-cell number and function. We have analyzed its mechanisms in human mature T cells, which have been thought to be insensitive until recently. Peripheral blood T cells showed sensitivity to GC-induced apoptosis soon after the proliferative response to a mitogenic stimulation, and were also sensitive to spontaneous (ie, growth factor deprivation-dependent) apoptosis. CD8+ T cells were more sensitive to both forms than CD4+ T cells. Acquisition of sensitivity to GC-induced apoptosis was not associated with any change in number or affinity of GC receptors. Both spontaneous and GC-induced apoptosis were increased by the macromolecular synthesis inhibitors, cycloheximide (CHX) and puromycin. A positive correlation between the degree of protein synthesis inhibition and the extent of apoptosis was observed. Interleukin-2 (IL-2) IL-4, and IL-10 protected (IL-2 > IL-10 > IL-4) T cells from both forms of apoptosis in a dose-dependent manner. Our data suggest that spontaneous and GC-induced apoptosis regulate the human mature T-cell repertoire by acting early after the immune response and differentially affecting T-cell subsets. PMID- 7492779 TI - Differential expression of integrins on human thymocyte subpopulations. AB - Integrins represent a candidate group of cell surface receptors that may control the homing and population of the thymus by T-cell precursors and the subsequent migration of developing thymocytes through the thymic architecture. We have used multiparameter flow cytometric methods to characterize the expression of several members of the integrin family (alpha 3 beta 1, alpha 4 beta 1, alpha 5 beta 1, alpha 6 beta 1, and alpha L beta 2) on thymocyte subpopulations and have correlated integrin expression with other well-defined thymocyte differentiation markers. alpha 4 beta 1 was expressed by all thymocytes, but expression was highest on CD4-CD8- double-negative (DN) cells, high on CD+CD8+ double-positive (DP) cells, and lowest on mature single-positive (SP) cells, alpha 3 beta 1, alpha 5 beta 1, and alpha 6 beta 1 were present on 13%, 63%, and 26% of thymocytes, respectively, with maximal levels of expression on DN and SP cells, and low levels of expression on DP cells. Simultaneous analysis of alpha 4 beta 1, alpha 5 beta 1, and CD3 expression suggested a pathway of T-cell differentiation in the thymus in which the majority of the DN cells were alpha 4 beta 1hi alpha 5 beta 1hi, the DP cells alpha 4 beta 1hi alpha 5 beta 1lo/-, and the most mature SP cells were alpha 4 beta 1int. The stage-specific expression of integrins strongly implies their functional involvement during T-cell maturation in the thymus. PMID- 7492780 TI - The insulin receptor substrate-1-related 4PS substrate but not the interleukin-2R gamma chain is involved in interleukin-13-mediated signal transduction. AB - Interleukin-13 (IL-13) induced a potent mitogenic response in IL-3-dependent TF-1 cells and DNA synthesis to a lesser extent in MO7E and FDC-P1 cells. IL-13 stimulation of these lines, like IL-4 and insulin-like growth factor-1 (IGF-1), resulted in tyrosine phosphorylation of a 170-kD substrate. The tyrosine phosphorylated 170-kD substrate strongly associated with the 85-kD subunit of phosphoinositol-3 (PI-3) kinase and with Grb-2. Anti-4PS serum readily detected the 170-kD substrate in lysates from both TF-1 and FDC-P1 cells stimulated with IL-13 or IL-4. These data provide evidence that IL-13 induces tyrosine phosphorylation of the 4PS substrate, providing an essential interface between the IL-13 receptor and signaling molecules containing SH2 domains. IL-13 and IL-4 stimulation of murine L cell fibroblasts, which endogenously express the IL-4 receptor (IL-4R alpha) and lack expression of the IL-2 receptor gamma subunit (IL 2R gamma), resulted in tyrosine phosphorylation of insulin receptor substrate-1 (IRS-1)/4PS. Enhanced tyrosine phosphorylation of IRS-1/4PS was observed in response to IL-4, but not IL-13 treatment of L cells transfected with the IL-2R gamma chain. These results indicate that IL-13 does not use the IL-2R gamma subunit in its receptor complex and that expression of IL-2R gamma enhances, but is not absolutely required for mediating IL-4-induced tyrosine phosphorylation of IRS-1/4PS. PMID- 7492782 TI - Administration of recombinant human granulocyte-macrophage colony-stimulating factor after chemotherapy regulates the expression and secretion of monocyte tumor necrosis factor (TNF) and TNF receptors p55 and p75. AB - Monocyte expression and secretion of tumor necrosis factor (TNF) and TNF receptors (TNF-R) p55 and p75 was studied in patients receiving granulocyte macrophage colony-stimulating factor (GM-CSF) after intensive chemotherapy. TNF expression and secretion of biologically active TNF was increased at regeneration compared with that of patients who had received chemotherapy alone. This effect persisted for several weeks after cessation of growth factor therapy. GM-CSF restored the responsiveness of monocytes to bacterial lipopolysaccharide (LPS), which appeared to be diminished after chemotherapy alone. Expression and secretion of TNF-R p55 and p75 by monocytes was augmented by GM-CSF therapy in association with the increase in TNF protein. We propose that GM-CSF administration after chemotherapy restores the normal responsiveness of monocytes to a secondary stimulus such as LPS and primes monocytes to respond to LPS with increased expression and secretion of TNF and TNF-R. PMID- 7492781 TI - In vitro and in vivo antileukemic activity of B43-pokeweed antiviral protein against radiation-resistant human B-cell precursor leukemia cells. AB - B-cell precursor (BCP) leukemia is the most common form of childhood cancer and represents one of the most radiation-resistant forms of human malignancy. In this study, we examined the antileukemic efficacy of the B43 (anti-CD19)-pokeweed antiviral protein (B43-PAP) immunotoxin against radiation-resistant BCP leukemia cells. B43-PAP caused apoptosis of radiation-resistant primary BCP leukemia cells, killed greater than 99% of radiation-resistant primary leukemic progenitor cells from BCP leukemia patients, and conferred extended survival to severe combined immunodeficiency (SCID) mice xenografted with radiation-resistant human BCP leukemia. Furthermore, the combination of B43-PAP and total body irradiation (TBI) was more effective than TBI alone in two SCID mouse bone marrow transplantation models of radiation-resistant human BCP leukemia. Thus, B43-PAP may prove useful in the treatment of radiation-resistant BCP leukemia. PMID- 7492783 TI - Characterization of T cells immortalized by Tax1 of human T-cell leukemia virus type 1. AB - Peripheral blood T cells were immortalized in vitro by introduction of the Tax1 gene of human T-cell leukemia virus type 1 (HTLV-1) with a retroviral vector and were characterized for transformation-associated markers. Long-term observation showed that these Tax1-immortalized T cells eventually exhibited very similar features that were characteristic of HTLV-1-immortalized T cells, ie, increased expression of egr-1, c-fos, IL-2R alpha, and Lyn and decreased expression of Lck and cell-surface CD3 antigen. Among these changes, an increase in the expression of Lyn and a decrease in the expression of Lck and cell-surface CD3 antigen were observed only in Tax1-immortalized T cells after long-term culture. The expression level of Tax1 protein did not differ significantly between early and late passage of cells, and the cellular clonality was found to be the same by the analysis of the retroviral vector integration site and the T-cell receptor beta chain gene rearrangement pattern. These changes in the expression of Lyn, Lck, and cell-surface CD3 antigen probably resulted from indirect effects of Tax1 that appeared after extended culture. PMID- 7492784 TI - Poor prognosis in multiple myeloma is associated only with partial or complete deletions of chromosome 13 or abnormalities involving 11q and not with other karyotype abnormalities. AB - Chromosomal abnormalities have major biologic and prognostic implications in leukemias. Cytogenetic information in typically hypoproliferative multiple myeloma (MM) is limited because of difficulties in obtaining analyzable metaphases. In this study, karyotypes and other known prognostic factors were analyzed in 155 newly diagnosed MM patients, entered on an intensive treatment program with two autotransplants. Complete remission (CR), event-free (EFS) and overall survival (OS) were analyzed using standard statistical methods. Abnormal cytogenetics were found in 39% of patients and were associated with a significantly lower CR rate (27% v 48%; P = .008). EFS and OS were inferior in patients with either partial or complete deletion of chromosome 13 or 11q abnormalities ("unfavorable" karyotype) when compared with the remaining patients (P < .001) who, as a group, had a similar prognosis irrespective of cytogenetic findings, ie, inevaluable, normal, or abnormal but without an "unfavorable" karyotype. The patients with abnormalities of both chromosomes 11 and 13 had a dismal prognosis with median EFS and OS of only 11 and 12 months, respectively. Significant associations were noted between an "unfavorable" karyotype and IgA isotype, elevated levels of beta-2 microglobulin (B2M, > or = 3 mg/L) and age > 60 years. On multivariate regression analysis, the absence of an "unfavorable" karyotype was the most significant variable associated with prolonged EFS and OS (P = .0001 and .0002, respectively). Other independent favorable variables were age less than 60 years, C-reactive protein (CRP) < or = 0.4 mg/dL and bone marrow plasmacytosis < or = 50% before treatment. On a multivariate analysis without cytogenetics, these same three standard parameters were identified as the only favorable variables. Patients not having all three standard favorable variables had a significantly lower CR rate (P = .03), EFS (P = .0001), and OS (P = .002) if an unfavorable karyotype was detected. We conclude that, in this program of uniformly treated MM patients, a poor prognosis was associated predominantly with abnormalities of chromosomes 11 and 13. PMID- 7492785 TI - The clonal nature of circulating Sezary cells. AB - To determine if circulating Sezary cells can be classified as reactive or neoplastic based on the ability to detect the presence or absence of clonal T cell receptor beta chain (TCR-beta) gene rearrangements by Southern blot analysis, we evaluated the peripheral blood of 25 patients: 11 patients with Sezary syndrome (SS), 11 with benign inflammatory dermatoses (BID), and three normal controls. Three of 11 patients with SS, with Sezary counts ranging from 14% to 52%, did not demonstrate any clonal TCR-beta gene rearrangements in the peripheral blood, despite a TCR-beta rearrangement by Southern blot analysis in the skin. Ten of 11 BID patients and all normal controls showed no evidence of a TCR-beta gene rearrangement in the peripheral blood. However, one patient with psoriasis demonstrated a TCR-beta gene rearrangement in the peripheral blood. The TCR-beta gene rearrangement detected in this patient, confirmed with polymerase chain reaction (PCR) amplification of the TCR-gamma gene rearrangement, did not correlate with the presence of circulating Sezary cells or the increased risk of neoplasia. Our results indicate that circulating Sezary cells may be monoclonal (neoplastic) or polyclonal (reactive), as defined by TCR gene rearrangement studies. Circulating Sezary cells in SS may be reactive in nature and not accurately reflect the actual tumor burden in the peripheral blood. The presence of circulating Sezary cells or the presence of a clone of cells defined by TCR beta gene rearrangement in the peripheral blood is not limited to neoplastic disease processes. PMID- 7492786 TI - High frequency of t(12;21) in childhood B-lineage acute lymphoblastic leukemia. AB - The recurrent t(12;21)(p12;q22) translocation fuses two genes, TEL and AML1, that have previously been shown to be independently involved in myeloid malignant proliferations. A search for rearrangement of the TEL locus in the region known to be involved in t(12;21) was performed by Southern blotting in a panel of hematopoietic malignancies. The presence of a t(12;21) was confirmed by fluorescence in situ hybridization (FISH) and/or reverse transcriptase (RT) polymerase chain reaction (PCR). We report that fusion of TEL to AML1 is specifically observed in at least 16% of the childhood B-lineage acute lymphoblastic leukemia (ALL) investigated, none of which had been previously identified as harboring t(12;21). PMID- 7492787 TI - Macrophage inflammatory protein-1 alpha receptors are present on cells enriched for CD34 expression from patients with chronic myeloid leukemia. AB - The response of normal and chronic myeloid leukemia (CML), CD34+ cells to human macrophage inflammatory protein-1 alpha (MIP-1 alpha or LD78) was assessed. In tritiated thymidine incorporation assays, stem cell factor plus granulocyte macrophage colony-stimulating factor stimulated thymidine incorporation in normal CD34+ cells was reduced to 72% of control values in the presence of MIP-1 alpha, whereas incorporation by CML CD34+ cells exposed to the same factors was not altered. In clonogenic assays, the presence of MIP-1 alpha gave a level of colony formation that was 71% of control values for normal progenitor cells, whereas for CML CD34+ cells colony formation was enhanced by 25%. These results suggest that, in vitro, CML progenitor cells are relatively refractory to the growth inhibitory effects of MIP-1 alpha. Using flow cytometry, the specific binding of a biotinylated human MIP-1 alpha/avidin fluorescein (FITC) conjugate to normal and CML mononuclear and CD34+ cell populations was quantified. The data indicate that (for both normal and CML CD34+ cells) there was a single population of cells that express cell surface receptors for MIP-1 alpha and this receptor expression was independent of cell cycle status. CML progenitor cells may be refractory to the effects of MIP-1 alpha as a result of events downstream from receptor expression. PMID- 7492788 TI - Growth and biologic properties of karyotypically defined subcategories of adult acute lymphocytic leukemia in mice with severe combined immunodeficiency. AB - Modest progress has been achieved over the past two decades in the treatment of adult acute lymphocytic leukemia (ALL). With modern therapy, response rates are 70% to 80%, but cure rates only average 25% to 30%. Improved in vivo models are needed to investigate the biology of adult ALL and to test new treatment concepts. Fresh leukemia samples from children with ALL have been successfully transplanted into mice with severe combined immunodeficiency (SCID), but no experience exists for adult ALL. We treated SCID mice with 2 mg cyclophosphamide 24 hours before intravenously injecting 20 x 10(6) viable leukemia cells obtained from 13 patients with newly diagnosed adult ALL within five defined phenotype/karyotype subcategories. Ten (76%) of 13 injected leukemia specimens representing all five categories engrafted. The median survival duration of mice was 20 weeks from the time of leukemia cell injection. The rate of engraftment by ALL subset was as follows: two of two T-cell, two of three t(11q23), two of two hyperdiploid, two of three t(9;22), and two of three diploid ALL. The pattern of organ involvement by leukemia in the mice was similar to that of the human disease. Immunohistochemistry and flow cytometry documented the stability of each leukemic phenotype after passage through SCID mice. Cells transplanted from the spleen and bone marrow of mice engrafted with ALL into recipient mice resulted in consistent engraftment. The survival duration in passage groups was similar to that in groups injected with primary cells. The high frequency of engraftment, availability of frozen original specimens, and successful passages in SCID mice provide an in vivo model of adult ALL suitable for further studies of the disease biology and for design of drug studies for the different subtypes of previously untreated adult ALL. PMID- 7492790 TI - The potential of iron chelators of the pyridoxal isonicotinoyl hydrazone class as effective antiproliferative agents. AB - Numerous studies have suggested that iron (Fe) chelators such as desferrioxamine (DFO) may be useful antitumor agents (Blatt and Stitely, Cancer Res 47:1749, 1987; Becton and Bryles, Cancer Res 48:7189, 1988). Recent work with several analogues of the lipophilic Fe chelator, pyridoxal isonicotinoyl hydrazone (PIH), indicate that some of these ligands are considerably more efficient than DFO both in terms of their Fe chelation efficacy and at preventing 3H-thymidine incorporation by neuroblastoma (NB) cells (Richardson and Ponka, J Lab Clin Med 124:660, 1994). Considering this fact, the present study was designed to test the antiproliferative effect of a wide range of PIH analogues to identify the most active compounds. A total of 36 ligands have been examined that were synthesized by condensation of three types of aromatic aldehydes (pyridoxal, salicylaldehyde, and 2-hydroxy-1-naphthyladehyde) with a range of acid hydrazides. The effects of these chelators were assessed using the human NB cell line, SK-N-MC. Although PIH was far more effective than DFO at preventing Fe uptake from transferrin, it was less effective than DFO at preventing cellular proliferation (DFO ID50 = 22 mumol/L; PIH ID50 = 75 mumol/L). In contrast, 14 PIH analogues were far more efficient than DFO at preventing proliferation (ID50 = 1 to 7 mumol/L) and may have potential as antitumor agents. The most effective compounds were those hydrazones derived from 2-hydroxy-1-naphthylaldehyde. Most of the PIH analogues were considerably more effective than DFO at both preventing 59Fe uptake from 59Fe-transferrin and in mobilizing 59Fe from prelabeled NB cells. In addition, a linear relationship between Fe chelation efficacy and antiproliferative activity was found only for hydrazones derived from salicylaldehyde. Apart from gallium (Ga) nitrate having an antiproliferative effect by itself, this metal potentiated the antiproliferative effect of PIH but not that of DFO. Spectrophotometric studies showed that PIH could chelate Ga, and it can be suggested that, like the PIH-Fe complex that donates Fe to reticulocytes (Ponka et al, Biochim Biophys Acta 718:151, 1982), the PIH-Ga complex may efficiently bestow Ga to NB cells. The results suggest that analogues of PIH deserve further vigorous investigation because they may be useful therapeutic agents for the treatment of cancer. PMID- 7492789 TI - Anti-B4-blocked ricin synergizes with doxorubicin and etoposide on multidrug resistant and drug-sensitive tumors. AB - Anti-B4-blocked ricin (anti-B4-bR) is an immunotoxin directed against CD19 positive cells that is currently being tested in several B-cell leukemia/lymphoma clinical trials. To explore the possibility of using anti-B4-bR in combination with chemotherapy protocols, we investigated the in vitro and in vivo cytotoxic effects of combining it with doxorubicin or etoposide using the lymphoma cell line Namalwa and a P-glycoprotein-expressing cell line, Namalwa/mdr-1, obtained by retroviral infection of Namalwa cells with the mdr-1 gene. Namalwa/mdr-1 cells were slightly more sensitive to anti-B4-bR than Namalwa cells; IC37 values were approximately 4 pmol/L and 8 pmol/L, respectively. When anti-B4-bR was combined simultaneously with doxorubicin or etoposide, additive to supra-additive killing of Namalwa and Namalwa/mdr-1 cells was observed. In xenografts of Namalwa/mdr-1 cells in severe combined immunodeficiency (SCID) mice, doxorubicin and etoposide at their maximum tolerated doses (3 mg/kg x 3 or 15 mg/kg x 3) showed no therapeutic effect. However, treatment with 5 daily bolus injections of anti-B4 bR (50 micrograms/kg) followed by treatment with doxorubicin or etoposide significantly increased the life span of the mice by 129% and 115%, respectively. After treatment with anti-B4-bR, the Namalwa/mdr-1 population expressed lower levels of P-glycoprotein, and this decrease may account for the synergistic action of the drug combinations. These results suggest that anti-B4-bR could be used to good effect in combination with current treatment regimens and further hint at a promising role for this immunotoxin in treatment of disease at the minimal residual disease stage, where cells may be resistant to chemotherapy. PMID- 7492791 TI - Increased cation permeability in mutant mouse red blood cells with defective membrane skeletons. AB - Cellular cation homeostasis in mouse erythrocytes with defective membrane skeletons was examined in three mouse mutants, hemolytic anemia (sphha/sphha), spherocytosis (sph/sph), and normoblastosis (nb/nb), and compared with reticulocytes produced by repetitive bleeding of congenic normal mice. To assess reticulocyte maturity, nucleic acid and transferrin receptor contents were measured by fluorescence flow cytometry; mutant cells were somewhat more mature than normal reticulocytes by these criteria. Red blood cell (RBC) sodium contents (Nac+) in homozygous sphha/sphha, sph/sph, and nb/nb animals were 30.1 +/- 0.9, 28.9 +/- 0.3, and 26.9 +/- 1.5 mmol/L cell, respectively, whereas cellular potassium (Kc+) was 102 +/- 2.6, 101 +/- 7.8, and 97.4 +/- 3.0. Nac+ and Kc+ in normal reticulocyte preparations were 11.3 +/- 0.7 and 123 +/- 10, respectively. Net Na+ and K+ fluxes in the presence of ouabain were markedly increased in mutant RBCs. Sodium uptake was 14.8 +/- 1.6, 15.4 +/- 3.3, and 14.7 +/- 3.1 mmol/L cell/h in sphha/sphha, sph/sph, and nb/nb mutants, respectively, whereas K+ loss was 17.0 +/- 4.0, 15.0 +/- 3.8, and 14.1 +/- 2.6. Normal mouse reticulocytes gained Na+ at a rate of 3.9 +/- 1.0 mmol/L cell/h and lost K+ at 6.0 +/- 2.1, rates indistinguishable from those in mature mouse RBCs. Potassium loss from sphha/sphha and nb/nb cells was not dependent on the presence of a Na+ gradient, and net cation movements were insensitive to bumetanide (sphha/sphha and nb/nb RBCs) and to chloride replacement with sulfamate (nb/nb cells). We conclude that mutant mouse RBCs with dysfunctional membrane skeletons have increased passive permeability to monovalent cations. These findings support a role of the membrane skeleton in the maintenance of the membrane permeability barrier and suggest that the abnormal permeability associated with human hereditary spherocytosis and elliptocytosis may be a consequence of the membrane skeleton defects reported in these disorders. PMID- 7492793 TI - Pyruvate kinase deficiency of mice associated with nonspherocytic hemolytic anemia and cure of the anemia by marrow transplantation without host irradiation. AB - Mutant mice with splenomegaly and nonspherocytic hemolytic anemia were found in an inbred colony of the CBA/N (hereafter CBA) strain maintained in the Japan SLC Haruno farm (Shuchi-gun, Shizuoka, Japan). The activity of pyruvate kinase (PK) in red blood cells (RBCs) of the anemic mutants decreased to 16.2% of normal (+/+) CBA mice. Because the mutant CBA mice showed a remarkable reticulocytosis (41.6%) and because the PK activity of reticulocytes is much higher than that of mature RBCs, the PK activity in mature RBCs of the mutant CBA mice was calculated to be 2.8% that of mature RBCs of CBA-(+/+) mice. Because RBC type PK is encoded by the Pk-1 locus of the mouse (chromosome 3), we designated the mutant locus as Pk-1slc. The anemia and PK deficiency of CBA-Pk-1slc/Pk-1slc mice were cured by bone marrow transplantation (BMT) from CBA-(+/+) mice. Prior irradiation was not necessary for the curative BMT. On the other hand, the BMT from CBA-Pk-1slc/Pk 1slc mice to nonirradiated CBA-(+/+) mice did not result in the decrease of RBCs and the reduction of PK activity. The present results indicate that CBA-Pk 1slc/Pk-1slc mice are a potentially useful animal model for studying pathophysiology of PK deficiency and for developing new therapeutic methods to correct PK deficiency. PMID- 7492792 TI - Evolutionarily conserved alternative pre-mRNA splicing regulates structure and function of the spectrin-actin binding domain of erythroid protein 4.1. AB - A developmental alternative splicing switch, involving exon 16 of protein 4.1 pre mRNA, occurs during mammalian erythropoiesis. By controlling expression of a 21 amino acid peptide required for high-affinity interaction of protein 4.1 with spectrin and actin, this switch helps to regulate erythrocyte membrane mechanical stability. Here we show that key aspects of protein 4.1 structure and function are conserved in nucleated erythroid cells of the amphibian Xenopus laevis. Analysis of protein 4.1 cDNA sequences cloned from Xenopus erythrocytes and oocytes showed that tissue-specific alternative splicing of exon 16 also occurs in frogs. Importantly, functional studies with recombinant Xenopus erythroid 4.1 demonstrated specific binding to and mechanical stabilization of 4.1-deficient human erythrocyte membranes. Phylogenetic sequence comparison showed two evolutionarily conserved peptides that represent candidate spectrin-actin binding sites. Finally, in situ hybridization of early embryos showed high expression of 4.1 mRNA in ventral blood islands and in developing brain structures. These results demonstrate that regulated expression of structurally and functionally distinct protein 4.1 isoforms, mediated by tissue-specific alternative splicing, has been highly evolutionarily conserved. Moreover, both nucleated amphibian erythrocytes and their enucleated mammalian counterparts express 4.1 isoforms functionally competent for spectrin-actin binding. PMID- 7492794 TI - Virucidal short wavelength ultraviolet light treatment of plasma and factor VIII concentrate: protection of proteins by antioxidants. AB - The use of solvent/detergent mixtures and various forms of heat treatment to inactivate viruses has become widespread in the preparation of blood derivatives. Because viruses that lack lipid envelopes and/or are heat resistant, eg, hepatitis A virus (HAV) or parvovirus B19 may be present, the use of two methods of virus elimination that operate by different mechanisms has been advocated. We now report on short wavelength ultraviolet light (UVC) irradiation for virus inactivation and enhancement of its compatibility with proteins by quenchers of reactive oxygen species (ROS). Treatment of an antihemophilic factor (AHF) concentrate or whole plasma with 0.1 J/cm2 inactivated 10(5) to > or = 10(6) infectious doses (ID) of encephalomyocarditis virus (EMCV), HAV, bacteriophage M13, vesicular stomatitis virus (VSV), and porcine parvovirus. However, the recovery of factor VIII was 30% or lower on treatment of an AHF concentrate and 60% on treatment of plasma. Factor VIII recovery could be increased with little or no effect on virus kill by addition of rutin, a flavonoid known to quench both type I and type II ROS. On treatment of plasma in the presence of rutin, the recovery of several other coagulation factors was also enhanced by rutin addition and typically exceeded 75%. Electrophoretic analysis of treated AHF concentrate confirmed the advantage of rutin presence; UVC irradiation of plasma did not cause discernible changes in electrophoretic banding patterns, even in the absence of rutin. We conclude that addition of UVC treatment to existing processes used in the manufacture of blood derivatives will provide an added margin of safety, especially for nonenveloped or heat-stable viruses. PMID- 7492795 TI - CD8-depleted donor lymphocyte infusion as treatment for relapsed chronic myelogenous leukemia after allogeneic bone marrow transplantation. AB - Donor lymphocyte infusions can reinduce complete remission in the majority of patients with chronic myelogenous leukemia (CML) who relapse into chronic phase after allogeneic bone marrow transplantation (BMT). Such infusions are associated with a high incidence of graft-versus-host disease (GVHD) and marrow aplasia. BMT using selective depletion of CD8+ lymphocytes from donor cells reduces the incidence of GVHD without an increase in leukemia relapse. We hypothesized that infusion of CD8-depleted donor peripheral blood lymphocytes could also reinduce complete remissions with a lesser potential to produce symptomatic GVHD in patients with CML who relapsed after allogeneic BMT. Ten patients with Ph(+) CML who relapsed a median of 353 days after BMT (range, 82 to 1,096 days) received donor lymphocyte infusions depleted of CD8+ cells. Nine patients received a single infusion and 1 received two infusions. Four patients were treated while in chronic phase with clonal evolution, 2 during accelerated phase, 3 during blast crisis, and 1 in a cytogenetic relapse. A mean of 0.9 +/- 0.3 x 10(8) mononuclear cells/kg were infused, containing 0.6 +/- 0.4 x 10(6) CD3+CD8+ cells/kg. Six patients achieved hematologic and cytogenetic remission at 4, 8, 11, 15, 39, and 54 weeks after lymphocyte infusion. Two patients developed > or = grade II acute GVHD, and 1 patient developed mild chronic GVHD. We conclude that donor lymphocyte infusions depleted of CD8+ cells can induce remissions with a low rate of severe acute GVHD in patients with CML who relapse after allogeneic BMT, supporting the hypothesis that CD8+ lymphocytes are important effectors of GVHD, but may not be essential for the graft-versus-leukemia effect against this disease. Further controlled studies are required to confirm these preliminary observations. PMID- 7492796 TI - Chloroquine treatment affects T-cell priming to minor histocompatibility antigens and graft-versus-host disease. AB - Graft-versus-host disease (GVHD) caused by T-cell recognition of minor histocompatibility (MiHC) antigens is a major complication of bone marrow transplantation. GVHD therapy has focused on removal or suppression of donor T cells, but modulation of MiHC antigen presentation to CD4+ T cells may represent an alternative approach. Chloroquine is known to inhibit major histocompatibility complex (MHC) class II presentation of antigen in vitro by affecting invariant chain dissociation from MHC class II. The goal of this study was to evaluate the role of chloroquine in abrogating T-cell priming to MiHC and GVHD in mice after transplantation of an MiHC incompatible donor. C57BL/6 mice were treated with phosphate-buffered saline or chloroquine at 400 micrograms intraperitoneally every day for 5 days before priming with BALB.B cells (MiHC-incompatible) followed by weekly injections of chloroquine at 400 micrograms for 4 to 8 weeks. Chloroquine treatment decreased the proliferative T-cell response to MiHC by 67% and the cytolytic T-cell activation by greater than 50%. After bone marrow transplantation (LP/J into C57BL/6; MiHC-incompatible), GVHD was significantly decreased in chloroquine-treated mice (17% with GVHD) as compared with that in controls (92% with GVHD). Chloroquine treatment did not have other effects in vivo on the normal T- and B-cell mitogenic responses, T-cell allogeneic responses, and MHC class II and I surface expression. Chloroquine treatment does decrease the ability of C57BL/6 antigen-presenting cells to stimulate C3H.SW T cells reactive with MiHC expressed on C57BL/6 cells, suggesting an effect on MHC class II presentation of MiHC in vivo. Treatment with chloroquine in vivo appears to result in decreased CD4+ T-cell priming to MiHC and GVHD by decreased class II MHC antigen presentation. Thus, chloroquine treatment may represent an alternative approach to control GVHD. PMID- 7492797 TI - In utero transfer of adult bone marrow cells into recipients with severe combined immunodeficiency disorder yields lymphoid progeny with T- and B-cell functional capabilities. AB - To determine if in utero transplantation could restore the immune system of mice with a severe combined immunodeficiency (SCID) disorder, C57BL/6Sz-scid/scid fetuses were injected on day 14/15 of gestation with adult congenic donor bone marrow (BM) cells. Congenic BM engrafted in one of eight (13%) recipients. Reconstitution of both lymphoid and nonlymphoid lineages was observed. In vitro and in vivo T-cell function was documented. Stem cells were shown to have engrafted by secondary transfer studies. When fully allogeneic C57BL/6 (H-2b) or B10.BR (H-2k) adult. BM cells were given to C.B-17-scid/scid (H-2d) fetal recipients, 15 of 54 (28%) recipients had evidence of engraftment, with up to 76% of peripheral blood (PB) being of in utero donor BM origin on day 131 postnatally. In all mice with persistent leukocyte engraftment, T- and B-lymphoid cells were entirely of donor origin. Donor T cells were tolerant to host but not third party alloantigens as measured in vitro. In vivo, T-cell function appeared intact. Although most mice had lower levels of B-cell engraftment than T-cell engraftment, mice with > or = 10% B cells were able to produce normal levels of IgM. Despite transplantation of fully allogeneic BM cells, stem cell engraftment could be demonstrated by secondary transfer of BM cells into lethally irradiated recipients that were congenic to the original in utero donor BM source. These data indicate that adult BM cells, even those fully allogeneic with the fetal recipient, can give rise to progeny with multilineage potential, which leads to restoration of T-cell and B-cell function. PMID- 7492798 TI - Therapy for ongoing graft-versus-host disease induced across the major or minor histocompatibility barrier in mice with anti-CD3F(ab')2-ricin toxin A chain immunotoxin. AB - A new pharmacologic agent, anti-CD3F(ab')2-ricin toxin A chain (RTA), was synthesized for the purpose of targeting T cells and as a means of treating established graft-versus-host disease (GVHD). The Fc region of anti-CD3 monoclonal antibody (MoAb) was removed to prevent its ability to activate T cells. The resulting F(ab')2 fragments were conjugated to deglycosylated RTA (dgRTA), a catalytic and potent phytotoxin. The resulting immunotoxin (IT) was potent (greater than 95% inhibition) and selective in inhibiting T-cell mitogenesis in vitro. In vivo, the IT depleted 80% of T cells in mice receiving bone marrow (BM) transplants. Transplantation in an aggressive acute GVHD model using C57BL/6 donor cells and H-2 disparate B10.BR recipients resulted in an infiltration of CD3-expressing cells and a median survival time (MST) of 20 to 30 days. A 5-day course of anti-CD3F(ab')2-RTA (30 micrograms/d intraperitoneally) beginning 7 days after GVHD induction was beneficial in treating established GVHD in these mice, as evidenced by significantly prolonged survival (MST, greater than 80 days), superior mean weight values, and improved clinical appearance. Neither intact anti-CD3, unconjugated anti-CD3 F(ab')2 fragments, nor a mixture of anti-CD4 and anti-CD8 MoAbs (which are highly effective in prophylactic models) were as effective. F(ab')2 fragments made from anti-Lyt-1 (the murine homologue of human anti-CD5) linked to RTA were also not effective, despite the fact that both anti-CD3F(ab')2-RTA and anti-Lyt-1F(ab')2-RTA had similar half lives of about 9 hours. The IT also increased MST in two aggressive models of GVHD across non-H-2 minor histocompatibility barriers, indicating that the usefulness of anti-CD3F(ab')2-dgRTA is not limited to a single-strain combination. This agent should be further investigated as an alternative to current strategies for treating steroid refractory GVHD. PMID- 7492800 TI - Paradoxical increase of painful crises in sickle cell patients with alpha thalassemia. PMID- 7492799 TI - DLA-identical bone marrow grafts after low-dose total body irradiation: effects of high-dose corticosteroids and cyclosporine on engraftment. AB - Previous studies found that marrow allografts from DLA-identical littermates resulted in survival of 60% of recipient dogs after an otherwise lethal dose of 450 cGy of total body irradiation (TBI), either because of successful allografts or autologous recovery after rejection of the allografts. Forty percent of dogs died with marrow aplasia after allograft rejection. The current study asked whether allogeneic engraftment could be enhanced and survival improved by treating allograft recipients with high doses of corticosteroids or with cyclosporine (CSP), administered either before or after transplantation. Five dogs in group 1 received corticosteroids beginning on day -5 and ending on day 32 after transplant. The starting dose was 12.5 mg of prednisone per kilogram orally twice daily. All five dogs rejected their allografts; three died early with marrow aplasia and two showed endogenous marrow recovery. Nine dogs received CSP from day -6 to day -1 before transplantation at a dose of 20 mg/kg/d intravenously administered in divided doses. All nine dogs rejected the marrow allograft; six died with marrow aplasia and three survived with endogenous marrow recovery. Seven dogs received CSP after transplantation at a dose of 30 mg/kg/d orally from day -1 to day 35. All seven had sustained allografts (two mixed chimeras and five complete donor-type chimeras) and became healthy long-term survivors without graft-versus-host disease. These results extend previous observations and confirm that grafts of marrow from DLA-identical littermates improved survival of dogs exposed to low but otherwise lethal doses of TBI. Additional therapy with high-dose corticosteroids administered peritransplantation and posttransplantation or CSP administered before transplantation neither enhanced the rate of allogeneic engraftment nor improved survival; however, CSP administered after transplantation resulted in successful allografts and event-free survival in all cases. PMID- 7492801 TI - Serum levels of interleukin-10 in B-cell chronic lymphocytic leukemia. PMID- 7492803 TI - Differential modulation of P-glycoprotein, the multidrug resistance mediator, by Cremophor in normal versus leukemic hematopoietic cells. PMID- 7492802 TI - Presence of the t(2;5) in Hodgkin's disease. PMID- 7492804 TI - Self-reported signs and symptoms in breast implant patients with novel antibodies to silicone surface associated antigens [anti-SSAA(x)]. AB - In research animals, the immune adjuvant activity of silicone in eliciting antibodies to associated protein antigens is now well established. In humans, the immune adjuvant activity of silicone remains controversial. Clusters of data from various research groups are beginning to define the boundary conditions for future large epidemiological studies. In that spirit, this manuscript reports pilot clinical data from a serological study conducted 3 years ago and reported previously in this journal. Sera from a self-selected symptomatic population of patients with breast implants were assayed for elevated concentrations of antibodies showing binding avidity to silicone surface associated antigens [anti SSAA(x)]. In that study of 249 patients, two distinct statistically significant subgroups were identified on the basis of the serological assay alone: patients without any elevated anti-SSAA(x) (negative) and patients with elevated anti SSAA(x) (positive) (p < 0.001). In this study, a clinical survey returned by 226 of those patients was correlated with the previously acquired serological findings. The 11 most common clinical complaints reported by the 199 anti-SSAA(x) negative patients and by the 27 anti-SSAA(x) positive were compared and their frequencies analyzed. The 199 anti-SSAA(x) negative patients, as a group, showed a lower frequency of a variety of signs and symptoms compared to the 27 anti SSAA(x) positive patients. Statistically significant differences were seen in three of the symptoms: fever, foot pain, and sleep disturbance (p < 0.05). In addition, the syndrome of fever in the absence of local chest pain was a significantly associated with anti-SSAA(x) positivity (p < 0.001). PMID- 7492805 TI - Intermittent micromotion and polyethylene particles inhibit bone ingrowth into titanium chambers in rabbits. AB - We performed a histomorphological and morphometric analysis of the effects of short daily periods of micromotion and phagocytosable particles of high density polyethylene (PE) on bone ingrowth into a 1 x 1 x 5 mm canal within a titanium chamber in rabbits. The micromotion chamber (MC) was implanted in the tibia of nine mature New Zealand white rabbits. After osseointegration and first harvest of tissue, 40 micromotions (amplitude = 0.5 mm) were applied daily at a rate of 1 Hz for a 3-week period. The tissue within the chamber was then harvested. For the second treatment, PE particles (10(8)/mL) were placed within the canal. The tissue in the chamber was harvested 3 weeks later. The next treatment was a 3 week rest period, in which neither micromotion nor particles were utilized; a harvest followed. The final treatment combined PE particles and micromotion, followed by a harvest 3 weeks later. Sections from control harvests contained extensive trabecular bone arranged longitudinally throughout the canal in a fibrovascular stroma. Micromotion produced longitudinally oriented fibrous tissue within the chamber. PE particles were associated with macrophages, surrounding and engulfing the birefringent particles. The combination of particles and micromotion produced a fibrous stroma laden with macrophages. PE particles and micromotion, alone or together, produced a similar effect in inhibiting bone ingrowth, compared to nonmoved chambers without particles. In this short-term experiment, no additive or potentiating effect of these two stimuli could be demonstrated. PMID- 7492806 TI - A novel drug delivery system for osteomyelitis using porous hydroxyapatite blocks loaded by centrifugation. AB - A novel drug delivery system for osteomyelitis was developed using porous hydroxyapatite blocks (HA-b) loaded with antibiotics by centrifugation. In the study, 10 mm3 HA-b was placed in a container and mixed with antibiotic solution; the antibiotic was then loaded into the pores of the HA-b by centrifuging at 1500 rpm for 15 min. Slow release of HA-b in both moist form and dried form (by heating at 160 degrees C) was tested after loading with the antibiotic arbekacin sulfate (ABK), 1-N-(S)-4 amino-2-hydroxybutyryl dibekacin. To estimate the concentration of antibiotic, both forms of HA-b were placed in 3 mL of phosphate buffered saline (PBS), which was replaced every 48 h. In both groups, which were loaded with 70 mg ABK per one block of HA (concn 0.5 microgram/mL) which is sufficiently high to control most pathogens, was maintained for 21 exchanges of PBS (after 42 days). Minimum inhibitory concentration for methicillin-resistant Staphylococcus aureus (MRSA), 3.13 micrograms/mL, was maintained until nine exchanges took place (after 18 days). The centrifugation method is simple, and dried ABK produced by heating loaded HA-b is particularly useful in clinical applications for osteomyelitis. PMID- 7492807 TI - Effects of precoating surface treatments on fatigue of Ti-6A1-4V. AB - Grit blasting is a common procedure of roughening surfaces to promote physical attachment of porous coatings, but it has been shown to reduce fatigue strength. Shot peening is known to increase fatigue strength by inducing compressive surface stresses; however, it is not known how subsequent grit blasting affects these benefits. This study examines the endurance limits, Se, of ELI grade Ti-6A1 4V specimens under rotating cyclic bending, including polished (control); belted and beaded; belted, beaded, and grit blasted; and belted, beaded, shot peened, and grit blasted. Belting and beading resulted in a slight increase in Se, grit blasting caused a 15% reduction in Se from polished. Fifty percent of this reduction was recovered when shot peening preceded grit blasting, suggesting that residual compressive surface stresses, induced by peening, were not eliminated by the blast process. Roughness averages and RMS values did not correlate with Se trends. SEM results showed classical fatigue fractures, consistent with surface crack initiation. PMID- 7492808 TI - Preparation of fibrin glue: a study of chemical and physical methods. AB - Concentrated fibrinogen was prepared from whole blood by cryoprecipitation or chemical precipitation and combined with thrombin to make fibrin glue (FG). Surgical applications of FG include control of bleeding, adhesion of tissues, and sealing of tissue defects. The purpose of this study was to compare cryoprecipitation (cryo) of fibrinogen to precipitation using ethanol, ammonium sulfate (AS), and poly(ethylene glycol) (PEG). Our results suggest that AS precipitation is as effective as cryo in yielding fibrin glues with high bond strengths and is more effective than ethanol and PEG precipitation. In addition, the volume of FG per milliliter of plasma is greater after AS precipitation than after a single freeze-thaw cycle. It is concluded that AS is an efficient means for preparing FG from autologous blood. PMID- 7492809 TI - Biological materials fixed with an epoxy compound: comparison of the effects with or without ionically bound heparin. AB - Biological materials have been used as prosthetic devices such as heart valves, vascular grafts, and pericardial patches. These biological materials have to be fixed with crosslinking reagents and sterilized subsequently before they can be implanted in humans. Recently, a new crosslinking reagent, epoxy compound, has been used to fix bioprostheses. In this fixation technique, heparin may be ionically bound on the tissue surface. It has been shown that the amount of heparin bound to the tissue surface is proportional to the quantity of protamine impregnated in the biological tissues. However, it is not known if the impregnation of protamine will affect the crosslinking density of the biological tissues. This study was designed to compare the crosslinking densities of the epoxy compound fixed biological tissues with or without heparinization. Fresh porcine aortic valves procured from a slaughter house were first impregnated in various concentrations of protamine sulfate (0, 0.5, 1.0, or 1.5%) for about 30 min. The porcine aortic valves were then crosslinked in a 4% epoxy compound solution (Denacol EX-313). The porcine samples were taken out at various elapsed fixation periods: 18, 25, 48, 72, 96, and 120 h. Finally, the crosslinked porcine aortic valves were heparinized in a 0.5% sodium heparin solution for about 1 h. The crosslinking densities of the porcine leaflet and the aortic wall of each sample were determined by measuring their shrinkage temperatures. It was revealed that the impregnation of various concentrations of protamine did not seem to significantly alter the shrinkage temperatures of the porcine leaflet and the aortic wall throughout the entire fixation process (p > 0.05).(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7492810 TI - Reinforced poly(L-lactic acid) fibres as suture material. AB - In this study, reinforced poly(L-lactic acid) (PLLA) fibers made by a dry spinning/hot-drawing process were evaluated for use as a suture. The initial tensile strength of the PLLA fibers was lower than the initial tensile strength of the commercially available sutures: PDS, Vicryl, silk, and Ethilon. However, after 12 weeks immersion in a phosphate saline buffer at 37 degrees C, PDS sutures have lower tensile strength than PLLA sutures and the tensile strength of Vicryl was unmeasurable because of fragmentation. Initially, PLLA fibers disintegrated into fibrils during degradation triggering an inflammatory response comparable to degradable multifilament sutures. However, the intensity of the inflammatory response against the PLLA fibers decreased and after 80 weeks implantation in the muscle layer of the abdominal wall of rats it was comparable to the one against Ethilon. The inflammatory response against Ethilon, which is considered to be nondegradable, increased in the same period, probably due to the change in shape. In practice, the handling characteristics of PLLA sutures are superior to the monofilament sutures like PDS and Ethilon and comparable with the multifilament sutures like Vicryl and silk. The knot security of PLLA sutures are expected to be better than the knot security of the monofilament sutures, but this remains to be investigated. It is concluded that dry-spun/hot-drawn (reinforced) PLLA fibers have the potential for use as long-term degradable suture material. PMID- 7492811 TI - Effect of corrosion products (neodymium iron boron) on oral fibroblast proliferation. AB - The biological effects of the corrosion products of neodymium iron boron (Nd2Fe14B) magnets are largely unknown. The aim of this study was to identify the types of corrosion product and to evaluate the effect of the corrosion products (CP) of Nd2Fe14B magnets on the proliferation of human oral mucosal fibroblasts. Uncoated Nd2Fe14B magnets were stored in saline at 37 degrees C for 6 months and the corrosion products collected. 100 microL of a cell suspension (human oral mucosal fibroblasts [14 x 10(4) cells/mL]) was aliquoted into 72 wells of a 96 well plate, the remaining plates receiving culture medium only. After 12 h incubation at 37 degrees C, each well then received 100 microL of either (A) culture medium, (B) 100% CP, (C) 50% CP, or (D) 0% CP. The plates were reincubated at 37 degrees C for a further 48, 96, or 144 h. Fibroblast proliferation was assessed using the methylene blue uptake/elution technique. The compounds in the corrosion product were examined using quantitative X-ray analysis. Statistical analysis (ANOVA, Bonferroni's test 0.05, SAS v 6.04), showed that at each time point, the cell numbers in groups B, C, and D were significantly lower than group A. Within groups B, C, and D no significant differences were found, despite the suggestion of a dose response effect. Fibroblast proliferation in the presence of corrosion products was significantly lower than with culture medium. Fibroblast proliferation did occur in the presence of 0, 50, and 100% CP. The actual corrosion products appeared to be salts of iron but 3.2% (+/- 0.6) of neodymium chloride (NdCl3) was found. PMID- 7492812 TI - Technical note: development of a model for study of in vivo bone strains in normal and microgravity environments. PMID- 7492813 TI - Comparison of diametral shrinkage of centrifuged and uncentrifuged Simplex P bone cement. AB - Reducing the porosity of bone cement substantially strengthens the cement in fatigue. However, concern has been raised that reducing the porosity of the cement would increase the shrinkage of the cement and thus compromise the cement bone interface. We measured diametral shrinkage of cement mantles prepared with and without centrifugation of the cement. Four cement preparations (centrifuged and uncentrifuged mixed with monomer at either room temperature or 0 degrees C) were used to form cement mantles of four different thicknesses (2, 4, 6, and 8 mm) around a stainless steel rod. We conclude that porosity reduction dramatically increases the fatigue strength of bone cement without substantially changing the diametral dimensions of the polymerized cement when studied in the shape and volume of a cement mantle that would be used for total hip replacement. PMID- 7492814 TI - Occlusion and stability of synthetic femoral canal plugs used in cemented hip arthroplasty. AB - The occlusion and stability of five synthetic plugs used to restrict the femoral canal prior to cemented arthroplasty was assessed. A model was employed consisting of a hollow wooden dowel to simulate the canal, with adapters fixed to both ends to accommodate cement insertion and pressurization, and to produce a closed distal cavity. Three different canal diameters within the range accommodated by the plug selected were employed to assess insertion force, distal pressure during both insertion and pressurization of acrylic cement, and plug migration and leakage for each device. There was a wide variation among plugs in the magnitudes of the force and distal pressure upon plug insertion. During cement pressurization, cement leakage and/or plug migration was noted in all trials, and either of these events resulted in pressure rises in the distal cavity. Most notably, differences of only 0.5 mm in the diameter of the canal resulted in marked changes in the occlusion and stability achieved. It is concluded that for the range of canal sizes anticipated intraoperatively, these synthetic plugs appear deficient to varying extents in their ability to occlude or remain stable in the intramedullary canal. PMID- 7492815 TI - Bioabsorbable polymers update: degradation mechanisms, safety, and application. PMID- 7492816 TI - [Psychosocial aspects of cancer screening]. AB - Psychosocial reluctance is a major obstacle to the success of mass screening. In France, several opinion polls demonstrated that the fear of cancer as a disease, the fear of a positive result and the fear of the possible therapeutic consequences were the major determinants opposed to screening despite information often known to the public. The psychological obstacles appear less important in the US population, except for minorities. Age seems to be a discriminating factor, indeed after 50 years, reluctance to screening is increased probably due to a generation effect influencing the meaning of the disease and the interest of screening. Site specific constraints also exist, in colorectal cancer, 30% of the people feel as an embarrassment the manipulation of the stools. If false positives are responsible for a supplementary anxiety, it doesn't last long and it doesn't influence the observation of future campaign. These psychosocial obstacles as a whole justify the need of converging information that must take into account cultural problems. The favourable influence of the general practitioner explains his essential role for the success of mass screening. PMID- 7492817 TI - [Positron-emission tomography: role of 18F-fluorodeoxyglucose (18FDG) imaging in oncology]. AB - There is a renewed interest in positron emission tomography (PET) using 18F fluorodeoxyglucose (18FDG) since the development of large field of view cameras and the capability of regional distribution of 18FDG. This method may help solve three types of problems in clinical oncology: tumor diagnosis and extension assessment, prediction of treatment response and follow-up (diagnosis of recurrences and complications). The aim of this paper is to review the literature in this field. This technique is mostly used for brain, lung, rectal and breast tumors as well as for sarcomas. It is possible to diagnose an anaplastic transformation of a low grade glioma since 18FDG uptake correlates with the histological grade. 18FDG plays another important role in the evaluation of the brain tumor response to treatment and of the secondary effects or sequelae of this treatment. This technique is also useful in breast carcinomas: diagnosis in the case of a dense breast, detection of lymph nodes or other metastases which could modify the strategy. One of the most established roles of 18FDG PET is the diagnosis of rectal tumor recurrences. Furthermore, future results will probably confirm its usefulness in lung carcinoma, for the diagnosis and for treatment evaluation. Lastly, it plays an important role in soft tissue sarcomas at all stages of diagnosis and treatment. The results of the literature still have to be completed. However, if the capability of predicting tumor response to treatment is confirmed, this method will play an important role in patient management and will modify treatment strategies. PMID- 7492818 TI - [Topoisomerase I inhibitors. Review of phase II trials with irinotecan (CPT-11) and topotecan]. AB - Topoisomerase I inhibitors are a new therapeutic class whose clinical evaluation began a few years ago; Irinotecan (CPT-11) gave interesting results in colon cancer; side effects were neutropenia, diarrhea, vomiting and a cholinergic syndrome. Topotecan was useful in lung and ovarian cancer; side effects were mostly hematologic. Undergoing studies concern dose optimization, mode of administration and therapeutic associations. PMID- 7492819 TI - [Paclitaxel (Taxol) and docetaxel (Taxotere): results of phase II trials in monochemotherapy]. AB - The taxanes, paclitaxel and docetaxel, are the two presents clinically available representatives of a cytotoxic class with a new mechanism of action: they enhance microtubule assembly and inhibit their depolymerization. Their activity has been demonstrated in ovarian, breast and lung cancers. Paclitaxel and docetaxel are also promising agents in the treatment of head and neck, gastric and pancreatic cancer. Neutropenia is the dose limiting toxicity. Currently, use of premedication allows to circumvent hypersensitivity reactions encountered earlier with paclitaxel. For docetaxel, measures to prevent fluid retention are essential. PMID- 7492822 TI - [Relations between hematological toxicity and total and free plasma levels of etoposide in daily oral administration]. AB - This study was undertaken in order to evaluate the inter- and intra-individual pharmacokinetic variabilities and their impact on the toxicity of oral Vepeside Sandoz administered daily for 21 days. The pharmacokinetic results confirmed the low bioavailability of this formulation (14% +/- 10%) and its large interindividual variability. Moreover, a great intraindividual variability was shown between day 1 and day 21. This fact can explain that the relationship between the relative decrease in neutrophil count and the pharmacokinetic parameters was observed only with either the mean area under the curve of concentrations versus time (AUC) or the mean residual concentrations (Cr). The determination of the fraction of plasma etoposide unbound, which ranged from 4.6 to 24.8%, improved the pharmacokinetic-pharmacodynamy relationship for AUC but not for Cr. This study showed the potential interest of etoposide drug monitoring. However, no dosage adjustment could be performed with this oral etoposide formulation because of its large intraindividual bioavailability. Since this study was performed, the Sandoz company withdrew this formulation, and replaced it by one identical with that available in other countries. PMID- 7492821 TI - [Cerebral germ cell tumors in adults. A retrospective study of 19 cases]. AB - The authors report on the retrospective analysis of 19 patients with primary cerebral germ cell tumors which were treated between 1965 and 1993. Median age is 18 years (extremes: 16-55 years). There were 16 men and three women. The location of the primary tumor was the pineal area in six patients, suprasellar and hypothalamic area in five patients and other areas in eight patients. The histological pattern was non seminoma in six patients, dysgerminoma in eight; however no histological sample was obtained in five patients who did not have any particular characteristics (either cytological abnormalities or elevated tumor marker level). Three patients were treated by surgery only, eight patients received exclusive radiotherapy and eight patients had first line chemotherapy and further cranial irradiation. One was lost to follow-up. Six of eight assessable patients with dysgerminoma are alive with non evolutive disease (NED) after 15 to 176 month of follow-up. One out of five assessable patients with non seminomatous tumor in NED (163 month of follow-up). Finally all five patients who have no histological subtyping are alive with NED at 24 to 138 months. The standard treatment of dysgerminoma is currently first line chemotherapy followed by relatively low-dose and limited irradiation; the standard treatment of non seminomatous cerebral germ cell tumor is chemotherapy, the study of which is warranted with the aim to decrease the toxicity and to increase the efficacy. PMID- 7492824 TI - [Second International Conference on Biology "Prevention and treatment of gastrointestinal malignancies" Koln (Germany), January 9-12, 1995]. PMID- 7492825 TI - [86th meeting of the American Association for Cancer Research "New drugs in cancerology" Toronto, (Canada), March 18-22, 1995]. PMID- 7492820 TI - [Hodgkin disease: should high-risk patients be selected and chemotherapy be intensified?]. AB - Death still occurs in Hodgkin's disease (HD) patients with advanced stages, but also in early stages. Therefore, the identification of the population of HD patients who would best benefit from an intensification of treatment is a prerequisite in all stages. The best known prognostic factors are bulky mediastinum, B symptoms, and visceral involvement in advanced HD and, in early HD, the number of nodal areas. No satisfactory combination of these factors can clearly identify a group of patients requiring high dose- and potentially risky therapy. Therefore only a very small subgroup of patients with advanced disease (less than 5% of them) calls for intensified regimens, which makes randomized trials very difficult. A large data base, accrued through a cooperative effort and the addition of early stages to advanced stages, would help to clarify the prognostic analysis. In addition, the patient population needs to be selected on the basis of the best therapeutic approach to-date. The analysis parameter should take into account only the failures due to the tumor progression as opposed to those due to a combination of tumor progression and early plus late toxicities. Three successive steps are required to achieve the goal of high-risk patient's treatment optimization. The first is to define, on a well-defined tumor-risk scale, the kind of results that are to be considered as "unacceptable" when up-to date conventional strategies are applied. The second is to infer the size of the "high-risk" population, in inverse ratio to the severity of the risk accepted for "normal-risk" patients. The third is to tailor the iatrogenic risk, ie the intensity of the high-dose treatment, to the size of the target population and to the anti-tumor benefit that is anticipated. Obviously, the definition of these parameters belongs to the medical decision-makers. Nevertheless, the availability of a quantified tumor-risk scale would allow to elaborate a reliable strategy in patients at risk for failure, lethal toxicity and unbearable health expenses. A collection of adequate recent trial is proposed through the Third International Symposium on Hodgkin's lymphoma in Koln (21-23 September 1995). This effort will be pursued through the International Data Base for Hodgkin's Disease (IDHD). Newer and more efficient strategies may then be designed. PMID- 7492826 TI - [Should cancer prevention become a form of treatment?]. PMID- 7492823 TI - [Desmoplastic small round cell tumors in adults: a new entity]. AB - Desmoplastic small round cell tumor is a recently recognized clinicopathologic entity with distinctive morphologic and immunohistochemical features. Moreover, specific cytogenetic and molecular characteristics have been described. We report here five new adult cases. The median age was 22 years. The clinical presentation was intra-abdominal in four patients and para-testicular in one patient. In patients with intra-abdominal disease, the initial surgery consisted of suboptimal debulking in three patients and only biopsies in the fourth. The patient with para-testicular tumor underwent an orchiectomy. Chemotherapy including cisplatin, etoposide, cyclophosphamide and doxorubicin or epirubicin (PAVEP/PEVEP regimen) was delivered after initial surgery in patients with intra abdominal presentation or at relapse in the patient with para-testicular tumor. Only one persistent complete response was obtained and no salvage treatment was active in the other patients who died of progressive disease. A review of the literature concerning the clinicopathologic, cytogenetic and molecular features of this new entity is exposed. PMID- 7492827 TI - [Prevention and chemoprevention in cancerology. A quantitative analysis of medical literature]. AB - Prevention is seen as a major concept of medical literature even distorted by two prisms: search through key-words and an unique referring: The Medline data bank. This topic is as important as "cancer": found in more than 8% of all the articles of this data base. Nine years have been studied (1985-1993) and the mean rise for the appearance of the word "prevention" is +25% a year. The topic "chemoprevention" is quantitatively unimportant: in 1993 0.07% of the whole corpus mentioned it, and as few as 0.8% of the prevention-related articles, however this topic has demonstrated a dramatic rise: a mean of +150% a year through the nine years studied (and +250% between 1992 and 1993). This topic of "chemoprevention" appears specially in connection with cancerology which seems to be a preferential field for this approach: more than 80% of the articles selected by "chemoprevention" refers also to "cancer". Finally, in the restrictive frame of "the upstream treatment", search on "hope" topic evokes a shift from early diagnosis to prevention. PMID- 7492828 TI - [Is breast cancer prevention by tamoxifen possible?]. AB - Breast cancer is a frequent disease especially in the Western world with an incidence that increases by 1% each year. Distressingly, even nowadays, 40% of patients with breast cancer will die of their disease. Postoperative adjuvant therapy and detection of non-palpable cancers by mass screening of women have improved patient survival but have not impinged upon the steady increase in incidence. Treatment before radiological detection could improve the chances of survival and even prevent the full-blown expression of the disease. The probability that a woman will develop breast cancer during her life-time is high but not evenly distributed. Knowledge of predisposing factors can help to identify those women subject to greater risk than the general population. The most important known risk factors are age, family history, factors related to ob gyn antecedents, lifestyle, benign hyperplastic breast disease with or without atypia. These various risks, are often associated and cumulative. A woman who has already had breast cancer has a 3 to 5 times greater probability of developing cancer of the contralateral breast than the average woman. The second breast is therefore at risk in the same way as are the women with predisposing factors. Many clinical studies have demonstrated the benefits of postoperative adjuvant tamoxifen (TAM) in improving the disease-free interval and survival of patients. An analysis of the site of relapse has interestingly brought to light a marked decrease in the percentage of cancers of the contralateral breast. In order to establish whether TAM could prevent or delay the onset of high-risk women without detectable signs of illness into groups receiving either TAM or placebo. Is this ethically acceptable? The answer to this question rests heavily on an objective evaluation of the unwanted and beneficial side-effects of TAM in order to weigh up all pros and cons before initiating a trial. PMID- 7492829 TI - [Cancer of the breast: primary prevention of breast cancer by tamoxifen, eligibility criteria, alternatives to English proposals]. AB - During the spring of 1993, UKCCCR prevention trial group proposed to the french medical community to participate into the international breast cancer prevention trial by tamoxifen set up in Great Britain. In January 1994 this proposal was eventually judged unacceptable in France because of lack of reflexion on primary cancer prevention, medico-legal risks, tamoxifen use risks and the eligibility criteria. A randomized trial could be designed and carried out in France to evaluate the place of tamoxifen in breast cancer prevention in the following settings: post menopausal patient with atypical epithelial hyperplasia or lobular in situ carcinoma. PMID- 7492830 TI - [Primary prevention of colorectal cancer]. AB - Results from case-control studies and prospective studies suggest that diet is involved in the causation of large bowel cancer either as initiator, promoter or inhibitor of carcinogenesis. Available data are not sufficient to serve as a basis for firm specific dietary advice. In the present situation it is attractive to investigate available hypotheses within the frame work of intervention trials. The adenoma appears to be one of the most appropriate end point of intervention studies. Several arguments indicate that the adenoma-carcinoma sequence is a multistep process. Colorectal cancer could possibly be prevented by intervening in the development of a small adenoma or in the growth into a large adenoma. Four intervention trials have been published so far. One of them suggest a protective effect of antioxidants vitamins on adenoma recurrence and three of them conclude to the absence of effect of these vitamins. A protective effect of lactulose on adenoma recurrence has also been suggested. Three studies are currently evaluating the effect of calcium supplementation on adenoma recurrence or growth of calcium supplementation, three studies the effect of fibre supplements, two studies the effect of antioxidants (one of them with calcium) and two studies the effect of diet intervention. The results of these studies will be available within three years. PMID- 7492832 TI - [Chemoprevention of cancers]. AB - Cancer chemoprevention needs carefully controlled clinical trials, definition of study populations and well selected chemopreventive agents. Randomized clinical trials are on going. We tried to summarize studies in the head and neck, lung, stomach, oesophagus, skin, bladder and cervix regions. PMID- 7492831 TI - [Chemoprevention of cancer of the prostate]. AB - Prostate cancer is an important problem of health care. Experimental models of chemoprevention, particularly involving retinoids derivates have been described. Clinical trials are going on in the United States to demonstrate the role of finasteride in the prevention of prostate cancer. PMID- 7492833 TI - [Medical adverse effects of chemoprevention: the example of tamoxifen]. AB - Tamoxifen, a synthetic antiestrogen widely used for the treatment of breast cancer, is also being proposed for the prevention of this cancer among women at high risk for the disease. Such an approach requires an objective and accurate evaluation, not only of the expected beneficial effects, but also of the potential iatrogenic side effects which could result from the administration of this drug to a population of healthy women. The present article summarizes our present knowledge which results from studies, all carried out on women with breast cancer: we review the side effects on the female genital apparatus, the ovarian and non ovarian endocrine side effects, the effects on cardio-vascular and thromboembolic risks, on bone metabolism and on ocular and hepatic side effects. The potential carcinogenic role is only briefly mentioned since it has already been the subject of a publication in this journal. PMID- 7492834 TI - [Methods and intervention in populations with high-risk for cancer]. AB - Prevention envisions either a reduction in the incidence of a disease through direct action against its causes, or a reduction of the consequences of that disease (especially mortality), by action against the process leading to clinical expression of the underlying pathology. As in therapeutic medicine, preventive measures must be thoroughly evaluated for efficacy and secondary effects before they become standard recommendations. Two levels of evaluation can be identified: evaluation of feasibility, acceptability, involving quality control of the proposed programme, evaluation of results according to previously defined and quantified objectives. In order to demonstrate that the risk-benefit ratio of a preventive strategy is favorable, results in the intervention group must be compared with those in the control group, just as is done in therapeutic trials. Further more, prevention applies generally to healthy subject ie those not needing medical care and for whom, in the majority of cases, there will be no direct benefit. As a consequence, careful ethical consideration must be given to such programme. In particular, calculating the risk-benefit ratio must take into account the mental and social well being of people whose lives will be "medicalized" as a result of such intervention. Measurement of the consequences of such preventive actions on the life style and mental health of the population subjected to them necessitates the development of specific tools adapted to such situations. Research in methodology in this area is essential. If a population with almost 100% risk for a disease can be identified (genetic risk for example), aggressive preventive intervention can be limited to only that group. It must be emphasized that identification of subjects at risk, no matter how reliable, is not an end in itself. It is only justified, on a medical point of view, if we can offer these individuals a preventive programme with demonstrated benefit. The primacy of collective benefit over individual interest mandates that preventive intervention be based on voluntary informed consent of the target population. An alternative to individual consent could be, in some occasion, a "collective" consent. PMID- 7492835 TI - [Public health problems related to chemoprevention of cancers]. AB - The chemoprevention sets down numbers of problems and requires certain number of conditions. First of all, one has to demonstrate the action's effectiveness by a randomised trial using the appropriate effectiveness indicators and to confirm the results of the trial by a pilot prevention's project. The action shouldn't have neither inconvenient nor major risks. In numerous prevention's action, the inconvenient are superior to the benefits. The action should be aware of respecting the major ethical aspects: respect the person's autonomy, don't to be harmful, respect the equity and be beneficial; it also has to avoid important psychological consequences. At last, the prevention's action has to appeal to cost effectiveness strategy. PMID- 7492836 TI - [Medical chemoprevention of cancers. From the point of view of social sciences]. AB - The orientation of the medical activity toward preventive strategies knows currently an important development. This movement shows various logic, which are: on one hand a rational step, a strategic choice in favour of prevention, and on the other hand a choice by elimination, when the other forms of intervention establish insufficient efficiency. Prevention had to follow the path of validity to acquire pertinence. The gold-standard is randomised blinded trials and it is this kind of procedure that has been proposed in the BCPT (Breast cancer prevention trial). By comparison with a classic therapeutic trial the main differences of prevention trial lay in: criteria of inclusion, analyses of impact larger than the simple efficiency (quality of life), on the focus on the risk (especially genetic) and the legal context. One of the main points is the question of risks induced by an intervention on healthy persons. In this context, even the passage of an uncertainty to a "certainty" concerning induced risks can not always suffice to close the controversy. Physicians cannot in this case, refer to norms and to models since this type of intervention is new. PMID- 7492837 TI - [Chemoprevention of cancers, the point of view of health insurance]. AB - Faced with scientific progress, social security must be concerned by ethics, especially by protection of personal integrity. Financial necessities lead to take into account the efficiency of public health interventions. But in front of potentially toxic drug, are we allowed to compare possible advantages with potentially serious risks for healthy women? For these reasons, social security takes a stand in favour of carefully controlled trials, focused on restricted target populations which are estimated at high risk of cancer. Research of side effects should be included in the follow of the patients and special caution should be taken to avoid premature spreading of this behaviour. About a financial contribution of social security, the National fund for health prevention, education and information (FNPEIS) could perhaps not support such a project, regarding to his experimental nature. CNAMTS-INSERM research contracts could be an alternative. PMID- 7492838 TI - [Methodology and legitimacy of French clinical trials in primary chemoprevention of cancers. The point of view of the National Federation of French cancer centers]. AB - In connection with the breast cancer prevention trial (BCPT) controversy, the "Federation nationale des centres de lutte contre le cancer" (FNCLCC) decided to clarify, with the help of various discipline experts, its position on chemoprevention trials. If in terms of inclusion, two approaches can be opposed: high relative risk versus high attributable risk, these trials share in common a considerable importance of the informed consent according to potentially prosecution. A particular attention should be given to the unambiguous definition of end-points and indicators, and to monitoring of the side effects (medical or not). Non medical actors would have to play significant interventions in the schedule, the follow-up and the evaluation of such protocols. Methodological difficulties have been pointed, for example: recruitment, observance, drop out and confounding factors. French specificity has to be taking into account for the choice of pathologies, kind of interventions and global evaluation non limited to the sole dimension of the medical efficiency. PMID- 7492839 TI - [Guidelines from the National Federation of Cancer Prevention Centers (FNCLOC) on trials of drug prevention of breast cancer by tamoxifen ]. PMID- 7492841 TI - Papers from the European Seminar on ELG and EPG, Edinburgh, July 1994. PMID- 7492840 TI - Laryngograph: speech pattern element tools for therapy, training and assessment. AB - The clinically based, real-time analysis of speech into physically definable elements, which are of direct perceptual and productive importance, has become more readily possible in recent years as the result of microprocessor developments. The combination of the acoustic signal of speech derived from a microphone together with the accompanying Laryngograph signal provides the basis for a highly reliable set of facilities. The paper describes methods and results for the analysis of voice, frication and timbre for both quantitative analysis and teaching and therapy using interactive visual displays. A brief discussion is given of links to work in stroboscopy, electropalatography and the associated use of additional sensors. Finally reference is made to a complete clinical work station combining these different facilities together with the quantitative analytical procedures of the speech pattern audiometer. PMID- 7492842 TI - Speech pattern audiometry for clinical use. AB - A new PC-based speech perception testing system ('speech pattern audiometer') is described which aims to provide a simple and efficient clinical tool to assess patients' ability to make use of the acoustic speech pattern information which is fundamental to speech perception. The speech pattern audiometer is likely to find its main application in audiology and speech and language therapy clinics with those who are deaf or who have developmental or acquired speech-perceptual disorders. This system constitutes a module of a complete speech and hearing assessment and teaching/therapy workstation. PMID- 7492843 TI - Variation in fold vibration during voiced obstruents in Russian. AB - A laryngographic analysis was carried out of vocal fold vibration produced during the supraglottal constriction for fricatives and stops. The subjects were a group of Russian speakers with normal voice production. Differences in vocal fold behaviour within this group of normal speakers were investigated. Changes in vocal fold vibration occurring during voiced fricatives and stops were evident from the Laryngograph waveform. Interpretation of the Lx waveform patterns suggests that partial opening of the vocal folds through the vibratory cycle may occur in some tokens. The evidence for this particular vocal fold behaviour remains inconclusive, however. Variations in vibratory patterns were found both between speakers and between utterances from individual speakers. PMID- 7492844 TI - Quality analysis of laryngography in a busy hospital ENT voice clinic. AB - The Portable Laryngograph provides a non-acoustic plot of wet mucosal contact at the glottis over the vocal fold vibrational cycle called the Lx waveform in Laryngograph terminology. Although well-equipped clinics gain most ENT requirements for investigation from the video endoscope, the nature of stroboscopy prevents the investigation over single glottal periods and many transitory problems may be missed. A high-quality Lx signal waveform will reflect any irregularities in mucosal contact and substantially complements the endoscopy examination. Yield, however, is a problem in the environment of an ENT hospital voice clinic. In this topic paper the criterion of maximum acceptable normalised noise energy (NNE) = -15 dB is used to assess the quality of 61 Laryngograph measurements. By this criterion a pass rate of 78% was achieved. This is considered to be good within an ENT voice clinic population where voice problems are in fact the dominant reason for referral. PMID- 7492845 TI - Electrolaryngography in the assessment and treatment of incomplete mutation (puberphonia) in adults. AB - Puberphonia, or incomplete mutation of the adult voice, is a condition which causes the sufferer considerable distress and which carries a powerful social stigma. It can usually be treated successfully by a few sessions of speech therapy. This paper describes how these sessions may be made more effective by using laryngography to provide immediate visual feedback to the speaker in the form of the Lx waveform. Fundamental frequency distributions, measured from conversational and formally read passages, are also presented. These demonstrate the substantial lowering of pitch and reduction of pitch irregularity achieved after voice therapy. It is demonstrated that the perception of normal male voice quality is not only a matter of pitch but also involves a shift towards a longer closed quotient of the male chest register Lx waveform pattern. PMID- 7492846 TI - Quantitative measures of the palate using enhanced electropalatography. AB - Electropalatography (EPG) is a useful tool for investigating tongue dynamics in experimental phonetic research and speech therapy. However, data provided by EPG are a two-dimensional representation in which all absolute positional information is lost. This paper presents an enhanced EPG (eEPG) system which uses digitised palate shape data to display the tongue-palate contact pattern in three dimensions. The palate shapes are obtained using a colour-encoded structured light three-dimensional digitisation system. The three-dimensional palate shape is displayed on a Silicon Graphics workstation as a surface made up of polygons represented by a quadrilateral mesh. EPG contact patterns are superimposed on to the three-dimensional palate shape by displaying the relevant polygons in a different colour. By using this system, differences in shape between individual palates, apparent on visual inspection of the actual palates, are also apparent in the image on screen. Further, methods have been devised for computing absolute distances along paths lying on the palate surface. Combining this with calibrated palate shape data allows accurate measurements to be made between contact locations on the palate. These have been validated with manual measurements. In addition, vocal tract areas in the oral cavity have been estimated by using the absolute measurements on the palate for a given contact pattern, and assuming a flat tongue profile in the uncontacted area. PMID- 7492847 TI - Categorising [s], [integral of] and intermediate electropalatographic patterns: neural networks and other approaches. AB - We consider two approaches to the interpretation of EPG data, using materials designed for the investigation of [s]-[integral of] assimilations across word boundaries, as in boss Shep. A contact index is based on electrode contact in different row groups, whereas a neural network is used to determine patterns of contact. Both approaches are speaker sensitive, reflecting the wide variation in [s] and [integral of] articulation between speakers. The advantage of the neural net analysis is that the investigator need not make any assumptions about the relative value of specific electrodes or groups of electrodes. We found that the 'hand-selected' index, in general, modelled linguopalatal movement for all degrees of [s]-[integral of] assimilation better than the neural net. However, the neural net very successfully characterised stable /s/ and /integral of/ and has the potential to give an accurate representation of intermediate assimilatory cases, if trained on a more varied input set than the rather limited one used in this experiment. PMID- 7492848 TI - EPG bidimensional data reduction. AB - A new method of electropalatographic (EPG) data reduction is proposed. It uses the so-called discrete cosine transform for representing an EPG pattern as a weighted sum of elementary 'pictures'. It is shown that several of these pictures can be considered as being articulatorily relevant. The respective contributions of these pictures to the spatial arrangement of the linguopalatal contacts are given by transform coefficients which are used here as a set of new EPG indices: the scaled sum of contacts index (SSU), the left-right asymmetry index (LRA), the alveolar-palatal index (APA) and the lateral-median index (LME). It is shown that the information extracted from an EPG pattern by means of these indices is maximised. Because of its robustness, this method could be usefully employed in studies on articulatory disorders. PMID- 7492849 TI - New developments in EPG3 software. AB - New developments in EPG3 software are described including: a three-dimensional display of the electrodes on the palate in the form of a wire frame representation; a facility for annotating EPG3 files; and a new data management system. Recent methods of EPG data reduction are also described, including numerical indices for representing tongue-palate contact profiles. A new clinical version of the Reading EPG system, EPG4, is introduced. PMID- 7492850 TI - Electropalatographic and perceptual analysis of the speech of Cantonese children with cleft palate. AB - This study used electropalatographic and perceptual analysis to investigate the speech of two Cantonese children with repaired cleft palate. Some features of their speech, as identified from the perceptual analysis, have been previously reported as being typical of children with cleft palate. For example, fricatives and affricates were vulnerable to disruption, and obstruent sounds were judged by listeners to have posterior placement. However, some apparently language-specific characteristics were identified in the Cantonese-speaking children. First there was a relatively high incidence of initial consonant deletion, and for one subject /s/ and /f/ targets were produced as bilabial fricatives. EPG error patterns for target lingual obstruents were largely similar to those reported to occur in English- and Japanese-speaking children. In particular, broader and more posterior tongue-palate contact was observed, and intrasubject variability was noted. There was also evidence of simultaneous labial/velar and alveolar/velar constriction for labial and velar targets respectively. The clinical implications of the findings are discussed. PMID- 7492851 TI - Co-articulatory variability and articulatory-acoustic correlations for consonants. AB - This paper is an investigation of articulatory-acoustic correlations differing in degree of articulatory constraint. Data on F2 and on dorsopalatal contact (electropalatographic, EPG) were collected for the sequences /iCi/ and /aC./ with seven Catalan consonants differing in place and manner of articulation (velarised /l/, /n/, /n/, /s/, /f/, /l/, /p/). These consonants are characterised by different degrees of tongue dorsum constraint depending on their production requirements (dorsals > non-dorsals, fricatives > non-fricatives, etc.). The results showed an inverse relationship between vowel-dependent coarticulation and the degree of consonant-dependent articulatory constraint. F2 and dorsopalatal contact size were found to be positively correlated across consonants and speakers, and across consonants for each individual speaker. Correlation values were much lower for each consonant across speakers. These findings are discussed in the light of the acoustic theory of speech production and possible clinical applications are suggested. PMID- 7492852 TI - A study of obstruent sounds in school-age children with speech disorders using electropalatography. AB - In this study EPG was used to investigate the articulatory characteristics of voiceless coronal obstruent targets /s, f, tf, t/ in ten children with functional articulation disorders. Perceptually based judgements of /s, f, tf/ revealed a variety of distortions, including palatal, lateralised and dental articulations. Their productions of /t/ were judged to be acceptable. From the EPG data, it was observed that individual children produced abnormal and often idiosyncratic tongue-palate contact for sounds which were heard as distorted. Some common trends were identified, for example, palatal fricatives and affricates were produced with constriction and a groove configuration in the posterior region of the palate. Lateralised articulations were generally associated with complete contact across the palate, increased tongue-palate contact and, for some children, incomplete lateral seal was observed. Place of articulation (as revealed by EPG) varied for coronal targets heard as lateralised. Some children produced these sounds with contact in the alveolar region, others had palatal contact and one child had contact in the velar region. These differences in place of articulation had not been identified from an auditory-based analysis. One child was heard to produce dental fricatives and affricates, and the EPG patterns were found to have an asymmetrical groove configuration in the anterior region of the palate. The clinical implications of the findings are discussed. PMID- 7492853 TI - An electropalatographic analysis of stutterers' speech. AB - Electropalatography (EPG) was used to investigate tongue-palate contacts from three adult stutterers during fluent and dysfluent speech and the results compared with normative data. Recordings were made of 22 read sentences, a reading passage and a conversational sample. The results showed that the lingual/palatal contacts of some phonemes during periods of stutterers' fluent speech consistently differed from that of the non-stutterers. Furthermore, stutterers produced greater intrasubject articulatory variability. Finally, during successive repetitions, the EPG patterns often became less similar to the target (or 'normal') configuration. PMID- 7492854 TI - Cross-linguistic study of lateral misarticulation using electropalatography. AB - This paper reports on some preliminary aspects of a collaborative cross linguistic study of normal and disordered Japanese and British English speech. The investigation compares lateralised productions of parallel s which are abnormal in the two languages. EPG and acoustic recordings were made of four Japanese and four British subjects. The EPG patterns were classified according to certain criteria, such as the presence or absence of complete constriction between the tongue and the hard palate, and the area and location of this contact. Findings revealed that lateralised articulations varied between individual speakers, but that Japanese and English productions were broadly similar. Acoustically, misarticulations in both languages were characterised by a lower frequency of peak energy than would be expected in normal productions. PMID- 7492855 TI - Using electropalatography to treat severe acquired apraxia of speech. AB - This paper reports on the use of electropalatography (EPG) in the treatment of a patient with severe acquired apraxia of speech. At 2 years post-onset and after a period of conventional speech and language therapy, the patient, AD, presented with severe speech production problems affecting articulation, phonation and resonance. Treatment using EPG is described and both the patient's progress and his continued limitations are outlined. EPG provided a valuable form of visual feedback for the patient and revealed and clarified aspects of oral movements for speech and non-speech activity which had been difficult to capture via auditory perception. PMID- 7492856 TI - EPG treatment of a child with the Worster-Drought syndrome. AB - This report of a case study of an unusual child aims to set the use of electropalatography (EPG) within the wider context of general communication therapy. It concerns a child of 12 years, diagnosed as having congenital suprabulbar paresis (Worster-Drought syndrome) who presents with severe developmental dysarthria; his speech is unintelligible, with hypernasality and glottalised articulation. His baseline EPG assessment patterns show minimal tongue-to-palate contact for all lingual obstruents, although he can demonstrate some tongue movement for non-speech skills, and has a slow but near normal swallow pattern. EPG therapy was used for tongue movements, but was adversely affected by his velopharyngeal insufficiency. PMID- 7492857 TI - The application of electropalatography (EPG) to the remediation of speech disorders in school-aged children and young adults. AB - Electropalatography (EPG) is one of a number of computer-based biofeedback techniques being applied increasingly and apparently successfully to the treatment of speech pathologies. This paper briefly examines the function of such biofeedback systems within the remediation process and looks in particular at the features of the EPG system which render it a useful therapeutic tool. Details are summarised of a project initiated to evaluate the use of EPG in the management of a large group of speech-disordered children and young adults. Successful applications of the technique are presented by examining the different types of motor speech skill which subjects acquired during intervention. These are summarised as the establishment of completely new articulatory patterns, the inhibition of abnormal lingual patterns, and the modification of temporal or spatial aspects of one or several existing patterns. Possible reasons for the success of EPG as a therapeutic technique, and issues concerning subject selection, are discussed. PMID- 7492859 TI - Quality control of DXA instruments in multicenter trials. AB - Dual-energy X-ray absorptiometry (DXA) has become the measurement of choice for multicenter trials with bone density endpoints. When performing DXA measurements with several different systems, it is important to implement a quality assurance program to guarantee that any observed density changes are real and not due to machine and/or operator variability. In this study, we present a series of procedures based on phantom measurements designed to monitor DXA instrument stability. Techniques for longitudinal evaluation of machine performance and cross-calibration of instruments are described. These procedures are then demonstrated using quality assurance data collected from a number of different DXA scanners. Together these methods provide a defined approach to instrument quality control. Though based primarily on the use of spinal phantoms, these procedures can be generalized for use in any multicenter DXA study. PMID- 7492858 TI - Study of the intraosseous vessels of the femoral head in patients with fractures of the femoral neck or osteoarthritis of the hip. AB - Bearing in mind earlier studies which established a link between arteriosclerosis and mineral loss, or fragility of the bones, and also our recent study showing that patients with arterial disorders of the lower limbs also suffered from osteoporosis, we carried out a histological study of the number and appearance of the intraosseous vessels and trabecular bone volume in the femoral heads of patients undergoing surgery for either fracture of the femoral neck or osteoarthritis of the hip. The number of thick-walled vessels, arterioles or arterial capillaries was significantly diminished in the femoral heads of patients with fractures of the femoral neck (p = 0.007). In addition, in the latter patients, arteriosclerotic vascular lesions (rupture of the internal elastic lamina, medial thickening and fibrosis) were more frequent than in patients with osteoarthritis of hip. The possibility that, through chronic ischemia, arteriosclerosis may lead to disturbance of bone remodelling and loss of the mechanical properties of bone has not been contradicted by these findings. PMID- 7492860 TI - Spinal bone mineral loss in estrogen-replete, calcium-replete premenopausal women. AB - After peak bone mass in women is attained, the benefits of increased dietary calcium or supplemental calcium are uncertain. In a longitudinal, 4-year study we have investigated the effect of calcium intake on bone mineral in a group of 41 premenopausal women, aged 38-42 years at entry. Skeletal density was measured four times during the 4-year follow-up; spinal trabecular bone density (STBD) was measured by quantitative computed tomography, and midradius bone mineral density (RBMD) was measured by single photon absorptiometry. At baseline, no differences in bone density were observed among subjects in the highest and lowest quartiles of habitual dietary intake. Overall, STBD declined -0.86 +/- 0.15% per year (p < 0.001), but RBMD did not decline. Total calcium intake (dietary calcium plus supplemental calcium) did not correlate with the rate of STBD loss. Serum estradiol level did not decrease during the study, and bone loss did not correlate with the mean estradiol level. We conclude that premenopausal women in the fifth decade lose about 1% of spinal trabecular mineral yearly, in spite of a normal serum estradiol level and ample calcium intake. PMID- 7492862 TI - Risk factors for hip fracture in a high incidence area: a case-control study from Oslo, Norway. AB - The aim of this population-based matched case-control study was to evaluate the effect of risk factors for hip fracture in Oslo, Norway, which has some of the highest incidence rates ever reported. The study population comprised all non institutionalized persons 50 years or older living in the catchment area of two Oslo hospitals, and cases were 246 patients admitted for hip fracture during a 1 year period. The controls were randomly selected from the study population, matched 1:1 for age and sex. Hip fracture was associated with lean body stature, smoking, low grip strength and decreased levels of physical activity, and inversely with length of education. In addition, hip fracture was inversely related to indicators of total food intake (number of meals per day, frequency of dinners, and slices of bread per day). A relation between hip fracture and low vitamin D intake was also suggested, whereas no association with dietary calcium intake was found. Finally, increased risk of fracture was seen in persons reporting two or more hospital admissions in the previous 2 years, and in those reporting weight reduction due to poor appetite during the previous year. In conclusion, the risk factor pattern for hip fracture was much the same in the elderly population of Oslo as previously described in other populations with a lower incidence of fracture. This study also indicates a relation between hip fracture and low food intake. PMID- 7492861 TI - Relation of height and weight to the regional variations in bone mass among Japanese-American men and women. AB - We examined the magnitude of regional variations in bone mass among elderly, Japanese-American men and women. All subjects had bone measurements at the calcaneus, and at the distal and proximal radius sites. A subset of the women had, in addition, spine bone mass measurements. To provide a common measurement scale, the bone measurements were converted to age- and sex-specific Z-scores. The Z-scores between pairs of bone sites were then subtracted to yield the differences in bone mass between bone sites (expressed in Z-score units). For most individuals the differences were less than 1.0 Z-score; however, 12%-20% of the differences were at least 1 Z-score apart. The most similar sites were the distal and proximal radius: different regions within the same bone. Among the other bone pairs, the calcaneus and spine were the most similar to one another. The magnitudes of the differences in bone mass were associated with height and weight. Heavier subjects, for instance, had greater calcaneus than radius bone mass measurements, and greater spine than radius measurements. The spine and calcaneus are more weight-bearing than the radius sites. Associations were observed up to 0.25 Z-score per 10 kg difference in weight. Height was associated with bone mass differences in an opposite direction to weight. Taller subjects had greater bone mass at the radius sites than expected from their calcaneus or spine bone measurements (0.1 to 0.2 Z-score difference per 5 cm difference in height). Bone width partly explained the associations with height; that is, adjusting the radius widths reduced the associations with height. Overall, our results indicate that small to moderate differences between bone sites were common among our study population, and that the magnitudes of the differences were associated with height and weight. PMID- 7492863 TI - A randomized, double-masked, placebo-controlled trial of chlorthalidone and bone loss in elderly women. AB - Employing a double-masked, prospective design, bone loss at three skeletal sites has been monitored among 113 postmenopausal women participating in a placebo controlled trial of the thiazide-like diuretic chlorthalidone for treatment of systolic hypertension. The mean duration of chlorthalidone use was 2.6 years, at doses of 12.5-25 mg/day. Compared with placebo use, chlorthalidone use was associated with significant reductions in annual bone loss rates. Non-use of chlorthalidone was associated with bone loss at the calcaneus (-0.56% per year) and the proximal radius (-0.91% per year); borderline bone gain was observed at the distal radius (+0.39%). In contrast, chlorthalidone use was associated with bone gain at the calcaneus (+0.44% per year) and the distal radius (+1.51% per year); proximal radius bone loss was significantly reduced to -0.32% per year. The average increment for three appendicular sites was +0.9% per year. These data support a causal relationship between chlorthalidone use and reduced bone loss. PMID- 7492865 TI - Accurate assessment of precision errors: how to measure the reproducibility of bone densitometry techniques. AB - Assessment of precision errors in bone mineral densitometry is important for characterization of a technique's ability to detect longitudinal skeletal changes. Short-term and long-term precision errors should be calculated as root mean-square (RMS) averages of standard deviations of repeated measurements (SD) and standard errors of the estimate of changes in bone density with time (SEE), respectively. Inadequate adjustment for degrees of freedom and use of arithmetic means instead of RMS averages may cause underestimation of true imprecision by up to 41% and 25% (for duplicate measurements), respectively. Calculation of confidence intervals of precision errors based on the number of repeated measurements and the number of subjects assessed serves to characterize limitations of precision error assessments. Provided that precision error are comparable across subjects, examinations with a total of 27 degrees of freedom result in an upper 90% confidence limit of +30% of the mean precision error, a level considered sufficient for characterizing technique imprecision. We recommend three (or four) repeated measurements per individual in a subject group of at least 14 individuals to characterize short-term (or long-term) precision of a technique. PMID- 7492864 TI - Stress distributions within the proximal femur during gait and falls: implications for osteoporotic fracture. AB - The rates of fracture at sites with different relative amounts of cortical and trabecular bone (hip, spine, distal radius) have been used to make inferences about the pathomechanics of bone loss and the existence of type I and type II osteoporosis. However, fracture risk is directly related to the ratio of tissue stress to tissue strength, which in turn is dependent not only on tissue composition but also tissue geometry and the direction and magnitude of loading. These three elements determine how the load is distributed within the tissue. As a result, assumptions on the relative structural importance of cortical and trabecular bone, and how these tissues are affected by bone loss, can be inaccurate if based on regional tissue composition and bone density alone. To investigate the structural significance of cortical and trabecular bone in the proximal femur, and how it is affected by bone loss, we determined the stress distributions in a normal and osteoporotic femur resulting from loadings representing: (1) gait; and (2) a fall to the side with impact onto the greater trochanter. A three-dimensional finite element model was generated based on a representative femur selected from a large database of femoral geometries. Stresses were analyzed throughout the femoral neck and intertrochanteric regions. We found that the percentage of total load supported by cortical and trabecular bone was approximately constant for all load cases but differed depending on location. Cortical bone carried 30% of the load at the subcapital region, 50% at the mid-neck, 96% at the base of the neck and 80% at the intertrochanteric region. These values differ from the widely held assumption that cortical bone carries 75% of the load in the femoral neck and 50% of the load at the intertrochanteric region. During gait, the principal stresses were concentrated within the primary compressive system of trabeculae and in the cortical bone of the intertrochanteric region. In contrast, during a fall, the trabecular stresses were concentrated within the primary tensile system of trabeculae with a peak magnitude 4.3 times that present during gait. While the distribution of stress for the osteoporotic femur was similar to the normal, the magnitude of peak stress was increased by between 33% and 45%. These data call into question several assumptions which serve as the basis for theories on the pathomechanics of osteoporosis. In addition, we expect that the insight provided by this analysis will result in the improved development and interpretation of non invasive techniques for the quantification of in vivo hip fracture risk. PMID- 7492866 TI - Comparison of areal and estimated volumetric bone mineral density values between older men and women. AB - We compared areal bone mineral density (BMD) of the total body (TBMD), antero posterior lumbar spine at L3 (APS), lateral spine at L3 (LS) and femoral neck (FN). In order to understand better the effect of gender-related size differences on BMD, we also compared the estimated volumetric BMD at L3 (VLS) and the femoral neck (VFN). Subjects were asymptomatic women (n = 22) and men (n = 44) with an age range of 58-79 years. BMD at each site was measured by dual-energy X-ray absorptiometry using a Hologic 2000 in array mode. Results of the statistical analyses (ANOVA) showed the men to have significantly greater BMD at all areal sites [APS, LS (p < 0.05); FN (p < 0.01); TBMD (p < 0.001)]. The two estimated volumetric comparisons, however, showed no gender differences. Results demonstrate how measures from areal BMD measures can be misleading when comparing groups of different size. In older men and women planar measures may overestimate gender differences in BMD. PMID- 7492868 TI - Patient dose in morphometric X-ray absorptiometry. PMID- 7492867 TI - Biochemical markers of bone turnover to monitor the bone response to postmenopausal hormone replacement therapy. AB - Hormone replacement therapy (HRT) prevents postmenopausal bone loss, and is therefore increasingly prescribed to prevent the development of postmenopausal osteoporosis. Because of individual differences in the response to HRT as well as problems with compliance, it has been debated how the skeletal response to HRT should be monitored. When estrogen production decreases at the menopause, a number of biochemical markers of bone turnover increase considerably in the order of 50%-100% from baseline. When HRT is instituted, the markers decrease again within the following 3-6 months. In the present prospective study we investigated whether the determination of biochemical markers of bone turnover may be useful for monitoring the skeletal effect of HRT. Seventy-six early postmenopausal women received HRT and 43 received placebo. The treatment period was 24 months and the women were followed with repeated bone mass measurements (every 3 months) which allowed calculation of the bone loss. Serum and urine samples were collected at 3, 6, 12 and 24 months. The placebo group lost a significant amount of bone mineral density in both the forearm and the spine (p < 0.001), whereas the HRT group did not. There was, however, a relatively large overlap of values between the HRT and placebo groups, especially in the spine. After 3 months' treatment the correlation between the changes in the markers and the bone loss was r = 0.59, and this value increased to r = 0.66 at 6 months and r = 0.76 and r = 0.77 at 12 and 24 months, respectively.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7492869 TI - Assessment of age and risk factors on bone density and bone turnover in healthy premenopausal women. PMID- 7492870 TI - Perth International Bone Meeting. Fremantle, Western Australia, 10-13 February 1995. Abstracts. PMID- 7492871 TI - Teenage sex and the health of the public: an octogenarian's view. PMID- 7492872 TI - Preventing adolescent pregnancy: in hot pursuit. PMID- 7492873 TI - Safe sex in the era of AIDS. AB - The term masturbation entered the English language in 1776 in the translation of Tissot's Treatise on the Diseases Produced by Onanism. Tissot linked semen conservation theory from antiquity with degeneracy theory and attributed degeneration and death from the social disease (syphilis and gonorrhea) to semen wastage not only in the social vice of promiscuity and prostitution, but also in the secret vice of masturbation. Nocturnal semen loss became designated as spermatorrhea, a new disease requiring treatment. In the nineteenth century the campaign against masturbation became a medical mania. It reached its apogee under John Harvey Kellogg, M.D., who invented Corn Flakes and other cereal and nut foods as meat substitutes to reduce all carnal desire and, hence, masturbation. The stigma on masturbation remains. It prohibits rational discourse on masturbation, and nourishes the perpetuation of fallacies regarding its effects. The imagery of a masturbation fantasy is also the imagery of the personal lovemap, which may be unorthodox, warped and distorted paraphilically. Masturbation might become societally endorsed as a public health policy to help contain the HIV epidemic of AIDS. Nonetheless, the President of the United States in 1995 dismissed his Surgeon General, Dr. Jocelyn Elders, for advocating so sane a policy. PMID- 7492874 TI - Male erectile disorder: a public health perspective. PMID- 7492875 TI - Alcohol-related sexual assault. PMID- 7492877 TI - Teaching human sexuality to medical students: a unique challenge. PMID- 7492876 TI - Bioethics in the news ... (with commentary) PMID- 7492878 TI - Mastectomy and sexual identity: the reconstruction of self-image. PMID- 7492880 TI - Religious belief systems, psychotherapy and sexual health. PMID- 7492879 TI - Risk management: physician practice acquisition under the anti-kickback law. PMID- 7492881 TI - Learning together as well as separately. PMID- 7492882 TI - There is very little European research on intensive care nursing. PMID- 7492883 TI - Pain assessment in acute myocardial infarction patients. AB - The purpose of this study was to investigate the pain intensity and duration, and to assess the pain control intervention, in patients with acute myocardial infarction (AMI). METHODS: Ninety-seven patients (18 diabetics and 79 non diabetics) admitted to the hospital with chest pain were included in this study. Pain was measured on the numerical rating scale (NRS) 0-10, where 0 means no pain and 10 unbearable pain. All patients were followed for 12 hours, after the last chest pain episode. The data were statistically evaluated with the Student's t test and chi square (chi 2). RESULTS: The pain in AMI patients with diabetes mellitus was lower in intensity (P < 0.002) and shorter in duration (P < 0.000) respectively, compared with the non-diabetic AMI patients. The intensity of pain in patients with an anterior infarction tended to be higher (P < 0.03) than in those with an inferior infarction. Finally, the systolic blood pressure fell significantly (P < 0.000) 90 min after admission. No other significant differences were found. PMID- 7492885 TI - A multidisciplinary audit of manual hyperinflation technique (sigh breath) in a neurosurgical intensive care unit. AB - Manual hyperinflation of the lungs (sigh breath) is a technique which is used to deliver a large volume of gas to the lungs. The purpose of this technique is to either maintain or improve the respiratory status of the patient. However, the volume of gas delivered is rarely measured, and therefore there is little evidence that hyperinflation occurs. Through the process of audit staff were able to examine the technique critically and improve it. With the addition of measurement tools a defined standard was developed against which performance was measured. To explore the effects of a particular technique it is essential that the delivery of that technique is standardised. This audit has enabled staff to achieve a standard technique and create a position from which research into its effects can now take place. PMID- 7492884 TI - Involving families in care within the intensive care environment: a descriptive survey. AB - A descriptive survey design was utilised within a general intensive care environment, to describe the attitudes of nurses and relatives towards the provision of care by relatives to their critically ill loved one. Triangulation was employed utilising a questionnaire comprising a Likert scale, a checklist of participatory care activities, open questions and biographical questions. The total nurse population of the intensive care unit (ICU) was surveyed. 27 questionnaires were returned (a response rate of 75%). Of a possible 45 relatives surveyed, 20 returned questionnaires (a response rate of 44.4%). A high proportion of both the nurse and the relative samples (96.3% and 85% respectively) indicated their agreement with the concept of involving relatives in the physical care of their critically ill loved one. The results highlighted issues of personal choice for individual lay involvement and adequate information for families to become involved. For the nurse sample the major themes emergent were the problems of role adaptation for nurses and families involved, and building relationships. For the relative sample the categories emergent were adapting to the demanding ICU environment and identifying the parameters of their new caring role. The study suggests benefits, to both nurses and relatives, of lay participation in physical care of critically ill patients on the ICU environment. PMID- 7492887 TI - Paracetamol poisoning: a case study. AB - Over a 1-year period, 24 patients have been admitted to one intensive therapy unit (ITU) with fulminant hepatic failure secondary to paracetamol poisoning. This article reviews one such case and the subsequent management which led to successful transplantation, concentrating on the clinical and nursing implications of the patient's care. PMID- 7492888 TI - Pain assessment by patients and nurses, and nurses' notes on it, in early acute myocardial infarction. Part 2. PMID- 7492889 TI - Do intensive care staff suffer more stress than staff in other care environments? A discussion. PMID- 7492886 TI - Nursing management of congestive heart failure. Part I. AB - Congestive heart failure (CHF) is a common clinical problem and in its advanced stage has a poor prognosis. Approximately 400,000 individuals develop heart failure each year (Parmley 1989). CHF can be caused by coronary artery disease such as myocardial infarction, ischemia and hypertension. Should heart failure occur as a result of acute myocardial infarction, a classification tool developed by Killip can be used to assess the degree of CHF. Killip's classification is organised into the following four categories: Class I: no heart failure Class II: mild to moderate failure Class III: acute pulmonary oedema Class IV: cardiogenic shock (Killip, as cited by Benz 1989). This system helps in organising clinical signs and symptoms of heart failure. Due to the fact that CHF is now the most common hospital discharge diagnosis for those over the age of 65, critical care nurses need to be able to recognise complications and intervene rapidly (Parmley 1989) in collaboration with medical staff. The purpose of this paper is first to discuss cardinal signs and symptoms and clinical data associated with CHF. Second, critical care nurses' management of CHF is organised around common nursing diagnoses. PMID- 7492890 TI - Meropenem (meronem, zeneca). AB - Meropenem is the latest in a series of antibiotics noted for their broad spectrum of activity. Is it an alternative to established antibiotic regimen in the treatment of infections in hospitalized patients? PMID- 7492891 TI - To see matters from the patient's view in several different intensive care units. PMID- 7492892 TI - Calcifying odontogenic cyst associated with compound odontoma: a study on undemineralized material. AB - In a minority of cases of calcifying odontogenic cyst (COC) it is possible to observe the formation of dental hard tissues in the cyst wall. The use of undemineralized sections has allowed an evaluation of the mineralized tissues normally lost with the use of demineralizing agents. All the dental hard tissues presented a high degree of morpho- and histodifferentiation. The histochemical staining for calcium salts (von Kossa) showed the presence of areas of low mineralization in the portion of the lesion, where the tissue maturation was not complete. In conclusion the appearance of the dental hard tissues in this case of compound odontoma arising in the cyst wall of a COC is similar to that already described in compound odontoma not associated with COC. PMID- 7492893 TI - [Calcifying odontogenic epithelial tumor--a clinical and morphological study]. AB - The authors describe the clinical and morphological study of a calcifying epithelial odontogenic tumor, localized in the right mandibular angle and in the ascending branch, in a 29-years-old male patient. It is emphasized that this benign and non invasive tumor presents poor clinical manifestations, a typical radiological aspect and a characteristic histomorphology, allowing the diagnosis. Polygonal epithelial cells, amyloidosis and calcified deposits are the distinctive traits of the histomorphology. PMID- 7492894 TI - Comparison of the clinical effectiveness of a single and a triple-headed toothbrushes in a population of mentally retarded patients. AB - Tooth brushing is a very simple and effective method for removing daily dental deposits and for preventing dental and periodontal diseases. However, it can cause considerable manipulative difficulties among some populations, e.g., young children, physically handicapped and mentally retarded patients. In order to test and compare the efficiency of a newly designed toothbrush (Superbrush), we have performed a pilot study on patients staying at the "Reine Fabiola Village No. 1", an institution for patients who are followed for mental retardation at different levels. 30 patients were included in the study, aged between 18 to 40 years. Among these 30, five had to be eliminated for their incapacity to follow the initial training in how to brush their teeth. The comparison was made with a normal single headed tooth brush, in a double blind trial, based on bleeding and plaque indexes, performed on 6 different teeth on day 0, 7 and 21. The results of this study indicate that there is no significant difference between the two types of toothbrushes with respect to the ability of plaque removing and gingivitis prevention, during the period of time of this study. However, the easiness of manipulating this newly designed toothbrush renders it a useful tool for the dental hygiene for this special part of the dental compromised population. PMID- 7492895 TI - Malignant fibrous histiocytoma of the head and neck. Case report. AB - Malignant fibrous histiocytoma (MFH) is a rare tumor included in the group of soft tissue sarcomas. Approximately 1-3% of these tumors affect the head and neck, with about 100 cases reported to date. A case affecting the retromolar trigone and involving the postero-lateral portion of the left hemi-tongue is reported. Etiological and pathological aspects are discussed. Diagnosis, treatment and prognosis of this neoplasm are reviewed. PMID- 7492896 TI - Langerhans cells in odontogenic cysts. A retrospective study based on 142 cases. AB - Dendritic cells with the ultrastructural and immunocytochemical characteristics of Langerhans cells are commonly identified in lymphoid organs and in keratinized and non-keratinized epithelia, such as oral mucosa. These types of cells were previously described in the epithelial wall of periapical odontogenic cysts. S 100 protein positive cells were demonstrated immunocytochemically in all of our 142 cases of odontogenic cysts (32 developmental cysts; 110 inflammatory cysts). Their tropism for squamous epithelia may explain their presence in the epithelial wall of odontogenic cysts. Their number is not always correlated with an inflammatory condition. PMID- 7492897 TI - Non Hodgkin's lymphoma of the maxilla as a first clinical manifestation of HIV infection. Report of a case. AB - Non-Hodgkin's lymphoma (NHL) of the oral cavity is rare. When it occurs, one should be suspicious of infection with the human immunodeficiency virus (HIV). NHL is the second most common malignancy associated with the acquired immunodeficiency syndrome (AIDS). This report describes the occurrence of a maxillary NHL as a primary clinical manifestation of HIV infection. PMID- 7492898 TI - Melatonin: implications at the oral level. AB - The strong relationship between the pineal gland and the immune system continues to establish itself more and more. In situations of a pinealectomy or in those of inhibition of the production of melatonin, a state of immunosuppression is produced which disappears when the patient receives this hormone. Likewise, melatonin counteracts the negative effects that immunosuppressive drug treatment produce over the immunity. It is due to these facts that one postulates the possibility of utilizing melatonin in primary and secondary immunodeficiency states as well as in cancerous immunotherapy. The action of this hormone over the immune system is carried out through the T-helper lymphocytes, lymphokines and also it seems through certain pituitary hormones. On the other hand, lymphokines such as gamma-interferon and interleukin-2, as well as the thymus, can modulate the synthesis of melatonin at the level of the pineal gland. Without doubt, all these actions have important consequences at the time of treatment of our high risk dental patients who have, in one way or another, an altered immunologic system. The altered immune system has direct implications in primary oral pathologies, or in others derived directly from our dental treatment. PMID- 7492899 TI - Antibacterial action of dental cements: an in vitro study. AB - The antibacterial activity of seven commercially available dental cements (Eugespad, Dentical, Dycal, Expaliner, PR. Scell, PR. Base Cement, PR. Lining Cement) against 1) bacterial species implicated in carious lesions or in dental plaque (Actinomyces israelii ATCC 10048, Actinomyces viscosus ATCC 19246, Streptococcus mutans ATCC 25175, Streptococcus sanguis ATCC 10557) and 2) bacterial samples of stimulated saliva was studied, in vitro, using a modification of the method of McComb and Ericson (1987). Dycal and Expaliner did not affect bacteria whereas the other dental cements displayed some antibacterial properties. Eugespad was the most active followed by PR. Base Cement + PR. Scell + Dentical and by PR. Lining Cement. Associated with mechanical and biocompatibility properties, these differences could be taken into account when choosing a dental cement for clinical use. PMID- 7492901 TI - Measurement error in air pollution exposure assessment. AB - The exposure of an individual to an air pollutant can be assessed indirectly, with a "microenvironmental" approach, or directly with a personal sampler. Both methods of assessment are subject to measurement error, which can cause considerable bias in estimates of health effects. If the exposure estimates are unbiased and the measurement error is nondifferential, the bias in a linear model can be corrected when the variance of the measurement error is known. Unless the measurement error is quite large, estimates of health effects based on individual exposures appear to be more accurate than those based on ambient levels. PMID- 7492900 TI - Enamel fluorosis in rat's incisor: S.E.M. and T.E.M. investigation. AB - Findings on the alterations taking place in the enamel have demonstrated that they are generally caused by a daily use of highly fluoridated drinking waters. According to that, the Authors have carried out an ultrastructural study on lower incisors of albino rats after administering for 60 days water with a fluorine concentration five times the normal one. The samples, studied under the S.E.M., showed a general slowing of both the deposition and the maturation phase as well as the presence of some hypomineralized areas even after eruption. All this suggested the possibility that the damages observed were not due to the direct effect of fluorine on the enamel, but to the interaction between fluorine and ameloblasts. The Authors have then carried out an ultrastructural study on the enamel organ using the S.E.M. The results showed the presence of a well-evident endoplasmic reticulum, the lack in dense granules during the secretion phase, the lack in ruffle ended webs during the modulation phase, and the mitochondrial damage in the ameloblast. All this could justify the slowing of the enamel mineralization caused by fluorine effect on the ameloblasts. PMID- 7492902 TI - Respiratory health associated with exposure to automobile exhaust. II. Personal NO2 exposure levels according to distance from the roadside. AB - We have conducted several studies to investigate the effect of automobile exhaust on respiratory symptoms. This study was designed to explore differences in personal exposure levels among residents of zones located varying distances from trunk roads with heavy traffic in Tokyo. Personal nitrogen dioxide (NO2) concentration levels for residents and NO2 concentrations inside and outside the residences of each study participant were measured during ten seasons over three years. Three residential zones were determined as follows: Zone A was 0-20 m from the roadside; Zone B was 20-150 m; and Zone C, a reference zone, was a residential district in a suburban area. Approximately fifty residents were selected as the subjects of NO2 measurements. Study participants were female, between 40 and 60 years of age, and nonsmokers. All participants used gas cooking stoves with electric ignition. Outdoor NO2 concentrations in Zone A were always the greatest among the three zones during the study periods, and those in Zone C were consistently the lowest. Personal exposure levels in Zone A were generally higher than those in the other zones, and concentrations in Zone C were the lowest during seasons when no indoor heating was used. The highest mean values for personal exposure levels in Zones A, B, and C were 63.4, 61.0, and 55.3 ppb, respectively. In analyses in which participants were stratified by heater type, the mean personal exposure levels in Zone A were the highest and the levels in Zone C were the lowest for participants without unvented heaters; differences of NO2 levels between Zones A and C ranged from 10.0 to 23.9 ppb. When there were no indoor NO2 sources except gas cooking stoves, both indoor and personal levels of NO2 were attributable primarily to motor vehicle exhaust. In contrast, the use of unvented heaters during the heating seasons could cause NO2 exposures comparable to those attributable to motor vehicles. PMID- 7492903 TI - Chronic respiratory symptoms associated with estimated long-term ambient concentrations of fine particulates less than 2.5 microns in aerodynamic diameter (PM2.5) and other air pollutants. AB - Seventh-Day Adventists (SDAs), nonsmokers who had resided since 1966 in the vicinity of nine airports throughout California (n = 1,868), completed a standardized respiratory symptoms questionnaire in 1977 and again in 1987. For each participant, cumulative ambient concentrations of fine particulates less than 2.5 microns (microns) in aerodynamic diameter (PM2.5) were estimated from airport visibility data. Long-term ambient concentrations of estimated PM2.5 in excess of 20 micrograms per cubic meter (micrograms/m3) were found to be associated with development of definite symptoms of chronic bronchitis between 1977 and 1987. Estimated mean concentrations of PM2.5 were associated with increasing severity of respiratory symptoms related to general airway obstructive disease, chronic bronchitis, and asthma. It was felt that the observed relationships, with the exception of the relationship between increasing severity of chronic bronchitis symptoms and PM2.5, could be due to surrogate relationships with other ambient pollutants. PMID- 7492904 TI - Estimating fine particulates less than 2.5 microns in aerodynamic diameter (PM2.5) from airport visibility data in California. AB - Methods were developed for estimating fine particulates less than 2.5 microns in aerodynamic diameter (PM2.5) from airport visibility data which detected seasonality and allowed for possible site- and season-specific regressions of PM2.5 on visibility. The methods were applied to nine airports in California in order to produce estimates of PM2.5 for the years 1966-1986 based on 1,767 paired PM2.5/visibility data points where PM2.5 had been measured at a monitoring station near the airport. General F-tests indicated that site- and season specific regression equations resulted in a statistically significant reduction in residual error. The split halves correlation for estimating PM2.5 from visibility over all areas was 0.82. The methods were used to estimate long-term concentrations of ambient PM2.5 for an epidemiological cohort of 1,868 individuals. PMID- 7492905 TI - A meta-analytical approach examining the potential relationship between talc exposure and ovarian cancer. AB - The concern that use of talc or talc-containing substances in the perineal region of women may subject them to an increased risk for ovarian cancer has become an important issue in the study of ovarian cancer. The purpose of this paper is to examine whether this concern, heightened by several epidemiological studies purporting to show an increased risk, is valid. Epidemiological studies examining the possibility of this relationship are reviewed, and meta-analyses of their results are performed. The conclusion reached herein is that the evidence regarding the risk of ovarian cancer associated with talc exposure is equivocal, and further examination of the relationship is required before a sound conclusion can be made. PMID- 7492906 TI - Methodology for selecting substances for the National Exposure Registry. AB - The National Exposure Registry was created in response to the pervasiveness of chemical contamination at the nation's waste sites and the relative lack of information on human health outcomes associated with long-term, low-level exposure to most of these substances. A ranking scheme was developed by the Agency for Toxic Substances and Disease Registry (ATSDR) to select the substances for which substance-specific subregistries of the National Exposure Registry would be developed. This scheme uses a general decision analysis approach that incorporates the most relevant and up-to-date data available on the substances found at sites known to ATSDR. There are currently four general subregistries (volatile organic compounds, dioxins, heavy metals, and radioactive substances) made up of persons exposed to specific primary contaminants, as selected by means of this ranking scheme. PMID- 7492910 TI - Safety and home care nurses: a study. PMID- 7492907 TI - Indices of TCDD exposure and TCDD body burden in veterans of Operation Ranch Hand. AB - Using responses from a questionnaire detailing herbicide exposure during service in Vietnam and information on job classifications, we investigated the relationship between 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD) body burden and self-reported exposure in enlisted members of Operation Ranch Hand, the United States Air Force herbicide spraying mission in Vietnam. We constructed three TCDD exposure indices from the questionnaire data: the number of days of skin exposure (DAYS), the percentage of skin area exposed (PCNT), and a combined index (SRI) which was the product of these and the concentration of TCDD in the herbicide. A fourth index (AFI) based on gallons of herbicide sprayed and the number of men on the job was also studied. The regression model most predictive of TCDD levels (R2 = 0.61) included job classification (divided into four categories), the number of days of skin exposure, percent body fat during the tour of duty, and relative change in body fat. A model with job classification alone had an R2 of 0.60. The four exposure indices were constructed to further explain TCDD exposure in the job classifications with the highest potential for exposure: Ranch Hand flight engineers and ground crew. In these two groups, days of skin exposure was the index most significantly associated with TCDD levels. Overall, the best index of exposure was the number of days of skin exposure to herbicide. PMID- 7492911 TI - Preceptorship: creating clinical and educational partnerships. PMID- 7492912 TI - Seniors serving seniors: volunteers promote healthy aging project. PMID- 7492913 TI - Ronald McDonald house: fulfilling a special need for over 10 years. PMID- 7492914 TI - New insights into the pathogenesis of Warthin's tumour. PMID- 7492915 TI - Development of a new invasion and metastasis model of human oral squamous cell carcinomas. AB - A new model was devised in order to establish an in vivo model for oral carcinoma that exhibits significant local invasion and metastasis. One hundred and fifty two nude mice had tumour cells from one of two established oral squamous cell carcinoma (SCC) cell lines (OSC-19 and OSC-20) implanted into the tongue or the oral floor via an intra-oral route and, as a control, the subcutaneous tissue of the back. The back tumours showed an expansive growth pattern, lacking significant invasion of surrounding tissues. In contrast, the tumours implanted into the tongue or the oral floor exhibited invasive growth and the histological appearance was similar to that of the original tumours. Moreover, regional neck lymph node and pulmonary metastases were observed in this model. Regional neck lymph node metastases were detected in 81.0% of mice implanted with OSC-19 cells and in 13.6% of mice implanted with OSC-20 cells. OSC-19 and OSC-20 cells showed pulmonary metastases in 9.5 and 9.1% of mice, respectively. These results suggest that this intra-oral implantation model is valuable in the study of the mechanism of invasion and metastasis of oral SCC. PMID- 7492916 TI - Feulgen hydrolysis profiles and acid-labile DNA in oral squamous cell carcinoma. AB - The full Feulgen hydrolysis profiles of healthy and malignant oral epithelial cells were compared by measuring the staining density of nuclei using microdensitometry after varying hydrolysis times. Malignant nuclei contained significantly increased levels of acid-labile DNA. The relative amounts of the rapidly hydrolysable fraction were compared after 5 min hydrolysis for exfoliating epithelial cells from healthy oral mucosa, healing chronic ulcers and squamous cell carcinomas. Although the latter exhibited a wide range of values, analysis of variance showed significant differences (P < 0.05) between healthy control and ulcer groups compared to oral carcinoma. The relative proportion of highly acid-labile DNA in malignant nuclei showed a significant positive correlation with mitotic score (P < 0.01) but no significant correlation with nuclear area. It was concluded that since the susceptibility of DNA to acid hydrolysis probably reflects functional differences in nuclear activity between cells, estimation of the highly acid-labile fraction may have diagnostic and/or prognostic value. PMID- 7492917 TI - Case-control study of oral dysplasia and risk habits among patients of a dental hospital. AB - Several studies have investigated risk factors for oral cancer but few have considered precancer. Records accumulated from 1975 to 1993 of dental hospital patients with histologically confirmed oral dysplasia provided the opportunity for a retrospective case-control study of the association between oral precancer and smoking tobacco and drinking alcohol. Seventy sets of case notes were available and each case was matched with records of a control subject, known to be free from dysplasia from another study, for birth date, gender and presumed ethnicity. The relative risk (OR) of having a dysplastic lesion for smokers compared with non-smokers, or ex-smokers for > 10 years, was 7.00. Logistic multiple regression revealed a dose-response relationship for tobacco dependent upon the level of cigarette consumption. Also subjects with moderate or severe dysplasia included a higher proportion of smokers than those with mild dysplasia. No overall increased risk from alcohol was found. However, the proportion of subjects who drank spirits was significantly higher among cases than controls. The study reaffirms the role of dental practitioners in identifying individuals at risk of mucosal disease, the importance of public education about the risk factors, and the necessity for counselling patients with precancerous lesions on avoiding further risk. PMID- 7492919 TI - Patient self-reported and clinician-rated quality of life in head and neck cancer patients: a cross-sectional study. AB - One to six years after treatment for head and neck cancer, 68 patients were asked to fill in a questionnaire at home with a subsequent examination at the out patient clinic of the Ulleval University Hospital. 50 patients (74%) both answered the questionnaire (the EORTC QLQ-C30, a diagnosis specific questionnaire module, the GHQ-20, and two questions from a population survey) and presented for examination where the clinician rated side effects after treatment, and filled in the Karnofsky Performance Status and the Spitzer's Quality of Life index. No patients had relapse or second primary tumours at the time of examination but a variety of post-treatment side effects were recorded. The patients appeared to report lower quality of life including more post-treatment side effects compared with the clinician's assessments. The cost-benefit of the standard clinical follow-ups in terms of increased survival may be questioned. Patients seem to need better support in coping with the physical and psychosocial problems which often appear. PMID- 7492918 TI - A preliminary investigation of an association between dental restorations and carcinoma of the tongue. AB - The potential association of dental restorations and tongue carcinoma was studied. We reviewed the available pretreatment dental records of 133 patients with carcinoma of the tongue seen at the British Columbia Cancer Agency between 1958 and 1992. 75 patients had teeth adjacent to the ipsilateral side of the cancer (involved side) and the contralateral (control) side of the tongue, resulting in 150 tongue/teeth pairings. Overall there was no significant association between the presence of dental fillings and tongue carcinoma, as the prevalence of bilateral dental restorations was high. Dental restorations were found in 6 patients on the involved side only. Only 1 patient developed a cancer adjacent to normal teeth with contralateral restorations. We feel that this observation merits more study. Meanwhile, careful assessment of dentition and aggressive treatment of dysplastic lesions are warranted. PMID- 7492920 TI - Well-being and coping in oral and pharyngeal cancer patients. AB - Patients with oral and pharyngeal cancer often suffer serious functional impairments. To achieve a broader understanding of the patients' situation it is important to include their appraisal of present functioning. There may also be other factors of importance in the evaluation of well-being such as personality and coping. 42 patients with oral and pharyngeal cancer answered the Oral and Pharyngeal Nursing Care Questionnaires 12 months after treatment. The questionnaires contain 25 conditions (grouped as Therapy- or Psychosocial-related Conditions) from three perspectives: perceived severity, received support and received information. Other questionnaires included in the study were the Sense of Coherence Scale, Somatic Anxiety Scale (from the Karolinska Scale of Personality) and Health Index. The patients were classified with regard to the extensiveness of their surgery. Severe disturbances related to therapy were significantly correlated to weaker sense of coherence (r = -0.34), more anxiety (r = 0.31) and to worse feelings about general health (r = -0.37). The corresponding data with regard to severe disturbances related to psychosocial situation were r = -0.44, 0.52 and -0.65, respectively. There was no significant correlation between perceived severity and surgical extensiveness. The support and information concerning Psychosocial-related Conditions were evaluated as less than those concerning Therapy-related Conditions. The level of support and information was not significantly correlated to sense of coherence, anxiety, general health or surgical procedure. The patient's subjective appraisal of the seriousness of the circumstances connected with oral and pharyngeal cancer and its treatment is an important parameter in the care of these patients. PMID- 7492921 TI - The oral health of long-term survivors of acute lymphoblastic leukaemia: a comparison of three treatment modalities. AB - Sixty-eight children who were diagnosed with acute lymphoblastic leukaemia (ALL) prior to age 5 years and treated with chemotherapy alone, chemotherapy plus 1800 cGy cranial irradiation (RT), or chemotherapy plus 2400 cGy RT were assessed clinically for overall dental health. All patients were at least 60 months in continuous remission. Dental caries were assessed by NIDR diagnostic criteria, oral hygiene was assessed by the modified Oral Hygiene Index, and gingival health was assessed by the modified gingival index of Loe and Silness. There was no significant difference in caries experience between the three groups nor with the normal population. Those patients that received 2400 cGy RT had significantly higher plaque and periodontal index scores than patients in the other treatment groups. The results of this study suggest that: (1) children with ALL treated with any of the described modalities are at no greater risk of developing dental caries than the normal population; and (2) patients receiving 2400 cGy prior to age 5 years are at greater risk of developing periodontal disease than patients treated with other central nervous system prophylaxis regimens examined in this study. PMID- 7492922 TI - Dietary intake in head and neck irradiated patients with permanent dry mouth symptoms. AB - Radiotherapy of the head and neck region, which includes the major and minor salivary glands in the radiation field, usually leads to temporary or permanent xerostomia. This may affect eating and increase the risk of inadequate intake of energy and nutrients. The aim of the present study was to investigate the effects of radiotherapy-induced xerostomia on energy and nutrient intake in individuals treated for malignancies in the head and neck region. The dietary intake of 24 patients with a low chewing stimulated whole saliva flow rate (< 0.5 ml/min) and in age and sex matched controls with normal flow rate (> 1.0 ml/min) was recorded for 7 days. The average daily energy intake was nearly 300 kcal lower in the irradiated patients with dry mouth symptoms than in the control group. The mean intake in the former group was 1925 kcal per day whereas the control group had an intake of 2219 kcal per day. Irradiated patients with dry mouth symptoms had significantly lower mean intakes of vitamin A, beta-carotene, vitamin E, vitamin B6, folacine, iron and zinc than those in the control group. There was also a lower intake of vitamin C, but this was not statistically significant. The intake of vitamins A and C exceeded or reached the levels recommended in the Swedish Nutritional recommendations, but the average intakes of fibre, iron, beta carotene, vitamin E, zinc, selenium, and iron did not reach recommended levels, in neither the experimental nor the control group. There was a slight positive correlation between energy intake and saliva secretion rate in the control group, but the energy intake was totally independent of variations in secretion rate in the irradiated patients with low secretion rate. PMID- 7492923 TI - A database for mucositis induced by cancer chemotherapy. AB - Ulcerative mucositis has become an increasingly important toxicity of antineoplastic therapy. In an effort to establish mucositis risk prediction for specific cancer chemotherapy regimens, a 25 field database was developed. This paper describes the rationale and methodology for creation of the database and instructions for access to it via the Internet. PMID- 7492924 TI - Alteration in the frequency, severity and duration of chemotherapy-induced mucositis in hamsters by interleukin-11. AB - Ninety-five young, male Golden Syrian hamsters were randomly divided into five equally sized groups. One group served as a placebo control while the animals in the others received one of four doses of interleukin-eleven (IL-11) twice daily given by subcutaneous injection beginning on the first day of chemotherapy (day 0) and continuing to day 14. Mucositis was induced with 5-fluorouracil using a standard regimen of 60 mg/kg, intraperitoneally on days 0 and 2 followed by superficial mucosal irritation on day 4. Animals were evaluated daily beginning on day 6. Mucositis was assessed using a standardised technique in which randomly numbered daily mucosal photographs were scored by three blinded independent observers at the conclusion of the experiment. IL-11 favourably affected the frequency, severity and duration of mucositis. This phenomenon appeared to be dose dependent. Hamsters receiving 30 and 100 micrograms per day of IL-11 demonstrated significantly (P < 0.05) lower mucositis scores than did either the control or animals receiving 3 or 10 micrograms per day, although the latter had marginal beneficial effects. Additionally, survival was significantly better for hamsters receiving higher doses of IL-11 (85%) compared to the placebo control (46%). IL-11 administration also favourably affected weight loss. While stimulation of platelet production was noted in animals receiving IL-11, a lack of difference in bone marrow cellularity between test and control animals suggests that the mechanism by which IL-11 modifies mucositis is mediated at the epithelial or connective tissue level rather than through the marrow. The kinetics of IL-11 alteration of mucositis induction supports such a hypothesis. Further investigation is currently underway to establish a definitive mechanism by which IL-11 protects the oral mucosa. PMID- 7492925 TI - The effect on cell phenotype of the mutagenic peptide of herpes simplex virus type-1. AB - The transforming region of the genome of herpes simplex virus type-1 (HSV-1) encodes a peptide that raises the mutation frequency of cells. To find the effect of this peptide on cell phenotype, three types of cells were transfected with a shuttle vector plasmid that expressed the peptide. When immortalised rat fibroblasts were transfected they rapidly became anchorage-independent with high efficiency, but were not tumorigenic in nude mice. When monkey kidney cells were transfected, five clonal cell lines were isolated, of which one became anchorage independent but was not tumorigenic in nude mice. When human oral keratinocytes were transfected they did not become immortalised. The peptide therefore induced some of the features of transformation in different cell types, but did not induce a malignant phenotype in any cell. This suggests that interaction with co factors would be necessary for the peptide to contribute to the development of oral cancer. PMID- 7492926 TI - Expression of tenascin in odontogenic tumours. AB - We investigated the expression of tenascin in a series of odontogenic tumours (n = 63) of epithelial and epithelial-ectomesenchymal origin by using immunohistochemical methods. A heterogeneity of expression of tenascin was observed in odontogenic tumours. The heterogeneity was most prominent in odontogenic tumours not forming calcified tissues. In these ameloblastomas and adenomatoid odontogenic tumours, tenascin was mainly localised at the epithelial tumour cell-mesenchymal tissue interface. In the calcifying epithelial odontogenic tumour, ameloblastic fibroma and odontoma, a widespread stromal immunoreactivity was observed which was, however, unreactive in the calcified masses. The stellate reticulum-like cells and granular cells of ameloblastoma also showed a positive immunoreactivity for tenascin. The results of the present study suggest that expression of tenascin in the stromal tissue of odontogenic tumours differs according to the potential of forming calcified masses by the tumour cells irrespective of tumour cell morphology. PMID- 7492928 TI - Cancer therapies for the 21st Century. 9th annual meeting of the International Society for Oral Oncology, 2-4 June 1994, National Institutes of Health, Bethesda, Maryland, U.S.A. PMID- 7492927 TI - Role of intralesional vinblastine administration in treatment of intraoral Kaposi's sarcoma in AIDS. AB - The purpose of this clinical study was to determine the effect of intralesional vinblastine administration on intraoral Kaposi's sarcoma (KS) in AIDS patients. One hundred and forty-four KS lesions in 50 HIV-positive homosexual males (mean CD4 count 64/mm3) were treated periodically with intralesional vinblastine injection (0.1 mg/cm2) until lesion resolution or no further reduction in lesional area. The most common lesion sites were: palate 56% (hard palate 42%, soft palate 14%); gingiva 22% (maxillary 15%, mandibular 7%); and maxillary tuberosity 6%. The mean lesion area was 4.6 cm2 (range = 0.1-35 cm2). Complete resolution occurred in 74%. The mean reduction in lesional area was 93% for all lesions. Lesions with only a partial response (26%) to vinblastine had a mean reduction in the lesional area of 69%. The mean number of treatments was 2.4 (range = 1-6). The recurrence rate was 26% with a mean disease-free period of 12.9 weeks. Recurrence rates were highest for nodular (40%) and purple macular lesions with focal nodularity (36%). The most frequent complications were transient pain (72%), superficial mucosal ulceration (22%) and transient paresthesia (12%). Intralesional vinblastine administration produced complete resolution in a substantial number of intraoral KS lesions and represents a well tolerated treatment regimen for localised control of intraoral KS lesions. Owing to a 25% recurrence rate, re-evaluation is necessary for treatment of recurrent and new Kaposi's sarcoma lesions. PMID- 7492929 TI - Stability of fixation: results of fundus-controlled examination using the scanning laser ophthalmoscope. AB - We evaluated the stability of fixation in 38 healthy eyes (N), 30 eyes with macular degeneration (AMD) and 20 eyes with juvenile macular dystrophy (JMD) during our newly developed automated full-threshold static perimetry using the scanning laser ophthalmoscope (SLO). The point of fixation was recorded for each stimulus presentation. After the examination, the mean fixation point (MFP) as well as the standard deviation from the MFP, the examination time, and the reaction time were evaluated. Healthy eyes did not show a mean SD of more than 0.7 degree from the center of gravity of fixation with one exception. There was only poor correlation between age and deviation from the MFP (r = 0.09 for N; r = 0.18 for AMD; and r = 0.18 for JMD), whereas the correlation between visual acuity (VA) and deviation from the MFP was slightly better (r = 0.22 for N; r = 0.34 for AMD; and r = 0.52; P < 0.02 for JMD). Eyes with functional impairment mostly showed instable fixation (deviation, > 3 degrees), which was observed even in two patients with only slightly diminished VA of 20/25. However, stable fixation was observed in isolated cases even when VA was reduced to 20/100. The deviation of fixation was more pronounced horizontally than vertically in normals (0.40 degree:0.31 degree) and in patients suffering from AMD (0.88 degree:0.68 degree), whereas eyes with macular dystrophy did not show this phenomenon (0.82 degree:0.82 degree). The results demonstrate that there is the possibility of stable fixation even in eyes with a decrease in VA due to morphological changes in the macula.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7492930 TI - Digital imaging of central serous retinopathy using infrared illumination. AB - When used with an infrared laser, scanning laser technology allows imaging of subretinal structures. In the indirect mode (scattered light), drusen, subretinal edema, and other changes in the retinal pigment epithelial complex can easily be visualized as pseudoprominent structures. This study was undertaken to determine the role of infrared imaging in central serous retinopathy. A total of 22 patients affected by acute central serous retinopathy were recruited for the study. All patients underwent fluorescein angiography (488 nm) and infrared imaging (788 nm) using a scanning laser ophthalmoscope (SLO 101; Rodenstock). The confocal mode was used for the fluorescein angiography, but the indirect mode was applied for infrared imaging. In all patients, serous detachment could be visualized by infrared imaging as pseudoprominent, oval-shaped structures. The size was at least one disk diameter and correspondend very well to the clinical appearance. In all patients, late-phase (20 min) fluorescein studies revealed the typical focal leakage with progressive hyperfluorescence. In six patients (27%) the extent of the serous detachment could be seen in a slightly hyperfluorescent rim. Digital image analysis showed that the extent of the serous detachment in the fluorescein angiogram was similar to the size of the thickened structures in the infrared image (r2 = 0.94). This initial study suggests that noninvasive infrared imaging can be a very useful indicator of serous detachment. Further studies regarding the influence of medical or laser treatment must be carried out to prove the clinical relevance. PMID- 7492931 TI - Automatic exposure control in retinal laser therapy. AB - Retinal laser spots are often over- or underexposed due to inhomogeneities of fundus pigmentation or variations in the radiation power impinging on the retina. We developed a method to control the exposure time of each individual spot of retinal laser photocoagulation in real time. The laser light reflected from the spot area is recorded by a detector. The intensity of this light increases during the exposure due to bleaching of the retinal pigment. If the reflected light reaches a predefined level, the laser apparatus is immediately stopped by a computer, which controls the whole process. The procedure was tested in enucleated porcine eyes. After verification of the proper function, a pilot study in a small collective of 19 patients (20 eyes, 25 sessions) with clear optical media was performed. In all cases, the apparatus worked safely. Visible indications for overexposed spots, particularly bleeding in extreme cases, did not occur. However, about 2%-5% of the spots were underexposed, mainly due to eye movements of the patient during the coagulation. Large variations in the automatically controlled exposure time confirm the difficulty in properly adjusting this parameter without such an automatic device. However, to be applicable to patients with opaque optical media, the method must be improved. PMID- 7492933 TI - Automated perimetry with bright and dark stimuli. AB - Conventional perimeters use bright stimuli. In this study, dark stimuli were also used. Dark stimuli are employed for testing due to their lack of effects of local scatter and are utilized with the purpose of preferentially stimulating the off system, but their presentation with optical methods is difficult. This problem is solved by the use of a computer monitor. The objective of this study was to compare directly measurements obtained with bright and dark stimuli. Therefore, the central 30 degrees of the visual field of ten subjects with no ophthalmologic pathology was examined with bright and dark stimuli (size 10- and 30-min-of-arc). Threshold estimation was performed with the 4/2 method. Concerning the 10-min-of arc dark stimulus, no measurement outside the central 15 degrees could be performed because of insufficient dynamic range (sometimes even the darkest stimulus was not seen). Comparing the results obtained using bright and dark stimuli of the same size, we could not find a significant difference in sensitivity for this age group. PMID- 7492932 TI - Macular damage following lightning strikes. AB - Two men with recent history of lightining strike were referred to our hospital. Both patients complained of metamorphosia in one eye and reduced visual acuity. Funduscopy revealed target-like alterations at the fovea. Fluorescein angiography showed window defects of the central retinal pigment epithelium in both patients. One patient developed an anterior subcapsular cataract. If the eye is part of the current-circuit, the melanin granules of the iris, pigment epithelium, and choroid might act as a resistor. The resulting accumulation of heat may lead to damage of the surrounding tissues. PMID- 7492934 TI - High-pass resolution perimetry and light-sense perimetry in open-angle glaucoma. AB - To determine the correlation between the results obtained with light-sense and ring perimetry in glaucoma, we examined 201 eyes of 107 patients with ocular hypertension or different stages of glaucoma using Octopus 1-2-3 and the Ophthimus ring perimeter. Statistical parameters describing the mean visual field defect ["global defect, GD," or "neural capacity, NC" (Ophthimus)] were significantly correlated (P < 0.0001). Statistical parameters describing the local deviations were significantly correlated as well ["loss of variance, LV" (Octopus) and "local defect, LD" (Ophthimus), P < 0.0001]. Correlation between the MD and the LV or LD was somewhat less significant. As compared with the MD or LV (Octopus) the Ophthimus parameters GD, NC, and LD were less often pathological. We conclude from our data that due to the good correlation between light-sense and resolution perimetry results, both perimeters are equally suited for the follow-up of glaucoma patients. Sensitivity to early glaucomatous damage seems to be higher in light-sense perimetry (Octopus 1-2-3). PMID- 7492935 TI - Scotoma perception in white-noise-field campimetry and postchiasmal visual pathway lesions. AB - The authors examined 59 patients (32 women and 27 men) suffering from homonymous hemianopia by white-noise-field campimetry (Tubingen Electronic Campimeter) and by conventional perimetry (Tubingen Automatic Perimeter or Tubingen Manual Perimeter) and by computerized tomography (CT) or magnetic resonance imaging (MR). In all, 56 patients showed neuroradiologically detectable cerebral lesions, which were superimposed in a reconstructed "reference brain". Of these 56 patients, 18 were not capable of perceiving any white-noise-field scotoma, although their conventional perimetric findings showed defects. Lesions of these patients were concentrated along the optic radiation and spared the primary visual cortex. The lesions of the remaining 38 patients with scotoma detectable in both methods were accumulated in area V1, the primary visual cortex. The lesions of 6 patients, who had had persistent scotoma perception for more than 2 years, were situated at the occipital pole. The lesions of 8 patients with vanishing scotoma were concentrated in more rostral areas. PMID- 7492936 TI - Eclamptogenic Gerstmann's syndrome in combination with cortical agnosia and cortical diplopia. AB - Cortical blindness is defined as a loss of vision due to bilateral retrogeniculate lesions (geniculocalcarine blindness). Gerstmann's syndrome is a combination of disorientation for left and right, finger agnosia, and profound agraphia, alexia, and acalculia. It is due to a lesion in the left angular gyrus, situated at the confluence of the temporal, parietal, and occipital lobes. We report on a patient who suffered from severe underdiagnosed eclampsia and who developed bilateral extensive medial temporal, parietal, and calcarine ischemic infarctions during an eclamptic fit. In addition, ischemia destroyed the left angular gyrus. The combination of these lesions led to Gerstmann's syndrome with additional cortical agnosia and cortical diplopia. For the first few months following the ischemic insult, the patient had been cortically blind. Thereafter, the patient slowly regained a visual acuity of 0.1 in both eyes. She then experienced monocular and binocular diplopia. Her ocular motility was normal; there was no phoria or tropia. Monocular and binocular diplopia slowly became less severe over the following year. Now, 2 years after the incident, the patient has a visual acuity of 0.2 in both eyes and no double vision. However, the handicapping symptoms of Gerstmann's syndrome, which make leading a normal life impossible, have persisted--the patient still cannot cope alone, mainly due to the severe disorientation for left and right. The picture of cortical agnosia, cortical diplopia, and Gerstmann's syndrome is a very rare combination. Visual recovery and rehabilitation in cortical blindness are severely affected and made difficult by the symptoms of Gerstmann's syndrome. In our case the reason for such a dramatic clinical picture was eclampsia, whose prodomes had not been diagnosed in time. PMID- 7492937 TI - Human immunodeficiency virus-related microvasculopathy and Kaposi's sarcoma: a case-control study. AB - Ocular microangiopathic syndrome including retinal and conjunctival abnormalities is frequently found in patients with human immunodeficiency virus type 1 (HIV-1) disease. Kaposi's sarcoma (KS) is the most frequent neoplasia found in patients with HIV-1 disease. We have recently reported a significant association between conjunctival microvasculopathy and KS in 117 patients with HIV-1 disease. The objective of the present study was to determine whether this association is existent when matched patients with and without KS are compared. A total of 22 matched pairs were obtained under consideration of the absolute CD4+ lymphocyte count, Walter Reed (WR) classification, gender, and serum levels of beta-2 microglobulin and neopterin. Conjunctival microangiopathy was determined for each eye by a standardized rating scale ranging from 0 to 5, allowing a reliable and valid quantification of conjunctival blood-flow sludging. The mean value obtained for conjunctival sludge was 1.8 (SEM, 0.4) for patients without KS and 3.2 (SEM, 0.3) for patients with KS, demonstrating a clinically and statistically significant difference between the two groups (Student's t = 3.0; P = 0.003). This difference was higher when patients with a CD4+ lymphocyte count exceeding 200/microliters were regarded. Similar factors or mechanisms may contribute to HIV-related conjunctival microvasculopathy and KS. PMID- 7492938 TI - Behcet's disease: epidemiology and eye manifestations in German and Mediterranian patients. AB - Behcet's disease (BD) is a systemic disease of unknown etiology, characterized by multiple organ involvement. Although its incidence is high in the Mediterranean countries and in Japan, BD is not often found in German patients. We therefore compared the epidemiological and clinical involvement of BD in 39 patients of German and of Mediterranean origin. The first manifestation was oral aphthosis in 72% of patients, followed by eye involvement (62%), skin lesions (25%), and arthritic problems (23%). Eventually, all our patients developed oral aphthosis. Whereas Germans had eye involvement in only 55% of cases (Mediterraneans, 71%), they came up with venous thrombosis or thrombophlebitis more often (64%) as compared with Mediterraneans (29%). The first ocular symptom was anterior uveitis in 87.5% of patients; the visual prognosis was poor due to occlusive retinopathy. Among 22 eyes of 12 patients, 16 had a visual acuity of 0.1 or less after 5 years of disease. We found a similar association with human leukocyte antigen B5 (HLA B5) in the German patients (66.7%) and in the Mediterranean group (54.4%). There was no significantly higher HLA-B5 association with the uveitis or arthritis type. These results demonstrate a few differences between the two ethnic groups but illustrate that in Germans the disease also has a poor prognosis. PMID- 7492939 TI - The heavy eye phenomenon: orthoptic and ophthalmic characteristics. AB - A series is presented of nine patients who demonstrate the heavy eye phenomenon. Illustrations of these patients show the salient features of the phenomenon, including hypotropia, exotropia and pseudoproptosis. The condition is summarized and the pathogenesis of the eye and lid positions are discussed. The indications for surgery and some examples with results are shown. The correct diagnosis may prevent unnecessary investigation of pseudoproptosis and pseudo third-nerve palsy. PMID- 7492940 TI - Multiple roles of platelet-activating factor in the nervous system. PMID- 7492941 TI - Platelet-activating factor is a synapse messenger and a modulator of gene expression in the nervous system. PMID- 7492942 TI - NMDA receptor-dependent and -independent immediate early gene expression induced by focal mechanical brain injury. AB - In the present study we analysed, by in situ hybridization, the effects of an extremely localized mechanical brain injury, obtained by the simple needle insertion (30 g) in rat hippocampus or cortex, on the expression of several immediate early genes (c-fos, fosB, c-jun, junB, junD, zif/268). When the needle is deepened into the hippocampus through the cortex, a simultaneous ipsilateral activation of all examined IEGs is observed in both the cerebral cortex and in the dentate gyrus of hippocampus. Maximal effects are detected between 30 and 60 min with the following rank order of induction: zif/268 > c-fos- > junB > fosB > c-jun > junD. On the other hand, when the penetration of the needle is limited to the cerebral cortex the activation of the IEGs (c-fos, fosB, junB and zif/268) spreads throughout the ipsilateral cortex but does not involve the hippocampal region. Systemic administration of ketamine, a non-competitive antagonist of N methyl-D-aspartate (NMDA) receptors, blocks IEG expression induced by brain injury in the cerebral cortex and in the hippocampal dentate gyrus. Pretreatment with the anticonvulsant diazepam, the anaesthetic urethane, or the muscarinic receptor antagonist scopolamine do not affect the injury-induced genomic response. An important regional difference in the sensitivity to the blocking effect of ketamine can be observed analysing the results regarding the zif/268 gene expression in the hippocampus. A clear induction of this gene by needle insertion can be detected both in the dentate gyrus and in the hippocampal layers. However, the dentate gyrus induction is completely blocked by the ketamine pretreatment, while the induction in the hippocampal layers is not affected by this NMDA antagonist. The zif/268 induction in the hippocampal layers is not blocked even if the intracerebroventricular administration of a non-NMDA glutamate receptor antagonist is associated to the systemic pretreatment with ketamine. This result represents the first observation of injury-induced neuronal genomic responses that are not critically dependent on the NMDA receptor activity. PMID- 7492943 TI - Differential expression of protein kinase C isoforms in glial and neuronal cells. Translocation and down-regulation of PKC isoforms in C6 glioma and NG 108-15 hybrid cells: effects of extracellular Ca(2+)-depletion. AB - Protein kinase C (PKC), the major receptor for tumor-promoting phorbol esters, consists of a family of at least 12 distinct lipid-regulated enzymes. We examined the expression and regulation of PKC isoforms in C6-glioma and NG 108-15 hybrid cells. Western blot analysis indicated that both cell lines express four PKC isoforms, PKC alpha, PKC delta, PKC epsilon and PKC zeta. The expression of PKC alpha and PKC delta in C6-glioma cells was more abundant than NG 108-15 cells, however, PKC epsilon in NG 108-15 was more abundant than C6-glioma cells in which PKC epsilon was almost undetectable. Treatment of both cells with TPA for 10 min resulted in the translocation of PKC alpha, PKC delta and PKC epsilon to the membrane fraction. When the intact cells were treated with Ca(2+)-free, EGTA containing physiological saline solution, the membrane bound conventional PKC alpha (cPKC alpha) was greatly reduced and cytosolic cPKC alpha was only slightly increased. However, neither membrane bound nor cytosolic new PKC delta (nPKC delta), nPKC epsilon and atypical PKC zeta (aPKC zeta) was affected by extracellular Ca2+ depletion. In this condition, the translocation of cPKC alpha, nPKC delta and nPKC epsilon induced by TPA still occurred, however, that of cPKC alpha was reduced more than in the normal condition. After long-term treatment (17 h) with TPA, cPKC alpha, nPKC delta and nPKC epsilon were down-regulated both in the cytosol and membrane. The phenomena of cPKC alpha were confirmed by measuring the PKC activity with histone as the substrate. From in vitro endogenous phosphorylation studies, a 31 kDa substrate protein phosphorylation in C6 glioma cell membrane and 31 and 26 kDa proteins in NG 108-15 cell membrane were increased in the translocation but disappeared in the down-regulation of PKC. PMID- 7492944 TI - Immunocytochemical localization of calmodulin in PC12 cells and its possible interaction with histones. AB - The subcellular localization of calmodulin, a multi-functional calcium-binding regulatory protein, was examined immunocytochemically in undifferentiated PC12 rat pheochromocytoma cells and cells differentiated with nerve growth factor (NGF) and dibutyryl cyclic AMP. In undifferentiated PC12 cells, diffuse immunostaining for calmodulin was observed in the cytoplasm, and weak, patch-like staining was found in the nucleus. In differentiated cells, intense immunostaining for calmodulin was observed in the cytoplasm, while nuclear immunostaining was still evident. Immunoreactivity for calmodulin was also observed along newly-formed neuritic processes, with strong staining in varicosity-like structures and growth cones. Using double-label immunochemistry, the relative intensity of immunostaining for calmodulin among the nuclei was found to correlate with the relative intensity of immunostaining for histones in the same nuclei. A comparison of a profile of 125I-calmodulin binding in the nuclear fraction from PC12 cells to that of immunoblotting for histones in the same fraction indicated that some of the histones are calmodulin-binding proteins in PC12 cells. These results show that the level and subcellular distribution of calmodulin are altered during the course of nerve cell differentiation and suggest the possibility that histones may function as major nuclear binding proteins for calmodulin. PMID- 7492945 TI - Lipid peroxidation inhibits acyl-CoA:-1-acyl-sn-glycero-3-phosphocholine O acyltransferase but not CTP: phosphocholine cytidylyltransferase activity in rat brain membranes. AB - In brain tissue in vivo peroxidized according to three model systems, we determined two microsomal enzyme activities involved in phospholipid biosynthesis. The first, short-term model, was based on the i.v. administration to normal rats, twice a day, for a period of 1 week, of a sonicated emulsion of a peroxidized mixture of phospholipids and linoleate (4:1, w/w; 500 mg/day; hydroperoxides: 200-250 nmol/mg lipid). The half-life time of the injected toxic lipid species in the blood circulation was about 1 h. At the end of the week's treatment, brain and liver malondialdehyde, conjugated diene and lipid hydroperoxide levels were significantly higher in treated rats than in the controls. The second model consisted of the acute injection of aqueous Fe2+ solution (50 mM) into lateral ventricles, and the collection of brain tissue 2 h later. The third model was based on two consecutive injections of hydroperoxylinoleate (1 mg each) into lateral ventricles over a period of 18 h, and the collection of brain tissue 2 h after the second administration. In brain microsomal membranes prepared from peroxide- or iron-treated rats, lysophosphatidylcholine acyltransferase activity exhibited a significant inhibition. On the contrary, in microsomal preparations derived from the short term model, CTP:phosphocholine cytidylyltransferase activity was slightly stimulated. Intraventricular injection of linoleate or linoleic acid hydroperoxide left this enzyme activity unchanged. The effect of in vitro membrane peroxidation on both microsomal enzyme activities was investigated. By using an Fe2+ (20 microM)-ascorbate (0.25 mM) peroxidation system, the residual acyltransferase and cytidylyltransferase activities were 80 and 72% of initial activity respectively. Significant dose-dependent inactivation of acyltransferase (maximum loss of 45% of initial activity) was seen when 0.1-10 mumol of photooxidized phospholipids were preincubated with 100 micrograms of microsomal membranes. Unoxidized or photooxidized phospholipids (1 mM) promoted a slight stimulation of cytidylyltransferase activity. Altogether, the results suggest a link between oxygen radical generation and the perturbation of the membrane structure in which the enzymes are located. PMID- 7492946 TI - Intrathecal dynorphin-A infusion in rat spinal cord causes energy depletion, edema and neurologic dysfunction. AB - The opioid dynorphin-A (dynA) is thought to contribute to the secondary injury process following spinal cord trauma although little is known about the biochemical mechanisms involved. In the present study, we have used a combination of magnetic resonance imaging (MRI) and spectroscopy (MRS) and hindlimb motor function tests to examine the effects of intrathecal dynA infusion on rat spinal cord. Infusion of 100 nmol of dynA (1-17) caused pronounced edema development as determined by MRI at 24 h after infusion. Infusion of 100 nmol of the dynA (2-17) fragment, which does not have any activity at opiate receptors, also produced profound edema whereas 100 nmol of the low potency kappa opiate receptor ligand dynA (1-8) or artificial CSF (ACSF) did not produce any edema. Both dynA (1-17) and dynA (2-17) produced significant hindlimb motor deficits at 24 h when compared to dynA (1-8) and ACSF (P < 0.05), but the deficits in the dynA (1-17) group were significantly worse than in the dynA (2-17) treated animals (P < 0.05). Similarly, mortality in the dynA (1-17) treated animals was significantly higher than in the other groups (P = 0.002). Phosphorus MRS demonstrated that the dynA (1-17) and dynA (2-17) treated animals also had a pronounced decline in high energy phosphates in the spinal cord 24 h after infusion. We conclude that dynA contributes to spinal cord cell death by causing metabolic failure and edema development. PMID- 7492947 TI - Melatonin stimulates brain glutathione peroxidase activity. AB - Exogenously administered melatonin causes a 2-fold rise in glutathione peroxidase activity within 30 min in the brain of the rat. Furthermore, brain glutathione peroxidase activity is higher at night than during the day and is correlated with high night-time tissue melatonin levels. Glutathione peroxidase is thought to be the principal enzyme eliminating peroxides in the brain. This antioxidative enzyme reduces the formation of hydroxyl radicals formed via iron-catalyzed Fenton-type reactions from hydrogen peroxide by reducing this oxidant to water. Since the hydroxyl radical is the most noxious oxygen radical known, induction of brain glutathione peroxidase might be an important mechanism by which melatonin exerts its potent neuroprotective effects. PMID- 7492948 TI - Evidence supporting membrane fusion as the mechanism of myelin basic protein induced insulin release from rat pancreatic islets. AB - In order to clarify insulinotropic effects of the myelin basic protein (MBP) we studied mode of association and distribution of MBP in the pancreatic islets and tested the insulin-releasing activity of various MBP peptides. Rat pancreatic islets were first stimulated in a static incubation with 10 microM bovine MBP (bMBP) at a substimulatory (3.5 mM) glucose concentration. The islets exposed to MBP released significantly more insulin and glucagon in a second incubation in the absence of added stimulant and in the presence of 11.5 mM arginine than the incubated, non-stimulated islets and islets initially stimulated with 15 mM glucose. Response to stimulation with 15 mM glucose in the second incubation by islets exposed first to MBP was impaired compared to incubated, non-stimulated islets. Immunoelectron microscopy showed that MBP had entered into the islet cells and associated with membranes of intracellular vacuoles, most of which represented enlarged, often fused insulin granules. MBP was also present at the islet edge and in the intercellular spaces. Of the purified MBP peptides of sizes of 4.8-13.6 kDa, produced from the digestion with brain acid proteinase and with pepsin and covering the entire bMBP sequence, only the large peptides (1-88, 9.8 kDa and 43-169, 13.6 kDa) stimulated insulin secretion significantly. Heterogeneous peptide mixtures, obtained from a time-course digestion of bMBP by myelin calcium-activated neutral protease, consisting of peptides of approximate molecular weights of 8-11 kDa and larger, also stimulated insulin release. The glucagon-releasing activity of MBP peptides was low and followed the same pattern as the insulin-releasing activity. The present results suggest that MBP-induced fusion of the membranes of hormone granules is involved in MBP-induced insulin release. The hormone-releasing activity of the large peptides in addition to that of the intact molecule is explained as being due to the ability of these peptides to associate with membranes. MBP-induced hormone release and related effects could be associated with neuropathological conditions such as stroke and multiple sclerosis. PMID- 7492949 TI - Protection by MK-801 against hypoxia-, excitotoxin-, and depolarization-induced neuronal damage in vitro. AB - Exposure of rat hippocampal slices to 12-min hypoxia produced only mild neuronal damage, as 72% of all slices recovered their CA1-evoked population spike following a 30-min recovery period. However, when this hypoxic insult was administered in the presence of 2.5 microM kainate or AMPA, only 6 and 15% of the slices, respectively, recovered their neuronal function. This enhancement of hypoxic damage by kainate could be attenuated in a dose-dependent fashion by the kainate/AMPA antagonist GYKI 52466 but not by the competitive NMDA antagonist APV. Unexpectedly, the noncompetitive NMDA antagonist MK-801 also attenuated the kainate- and AMPA-enhanced hypoxic neuronal damage and was more efficacious than GYKI 52466. Considering (1) the ability of MK-801 to antagonize hypoxic neuronal damage in the absence or the presence of NMDA, kainate or AMPA; (2) the antihypoxic effect of MK-801 in the presence of APV + 7-chlorokynurenate, a pairing that supposedly blocks MK-801 binding to the NMDA receptor; (3) the ability of MK-801 to protect hippocampal slices against brain damage induced by depolarization + excitotoxin (50 mM KCl + mM glutamate for 60 min); and (4) the ability of diltiazem, an L-type calcium channel blocker, to protect hippocampal slices against hypoxic neuronal damage, we conclude that the mode of action of MK 801 cannot be explained by its NMDA receptor antagonistic properties alone. A possible blockade of Ca2+ channels, most likely of the L-type, by MK-801 should be considered along with other mechanisms. PMID- 7492951 TI - Inhibition of [3H]mebendazole binding to tubulin by structurally diverse microtubule inhibitors which interact at the colchicine binding site. AB - A rapid and convenient radioligand assay was used to characterise the interaction of several structurally diverse microtubule inhibitors with the colchicine binding domain of tubulin. Values determined for the inhibition of [3H]mebendazole binding to tubulin by colchicine, combretastatin A4, NSC 181928, NSC 321567, podophyllotoxin and tubulozole-C provided an independent measure of the relative potency of these compounds. This methodology has several advantages over the inhibition of [3H]colchicine binding as a technique for investigating the molecular mechanisms involved in determining tubulin-ligand interactions. PMID- 7492950 TI - In vitro oxidation of vitamin E, vitamin C, thiols and cholesterol in rat brain mitochondria incubated with free radicals. AB - The kinetics of oxidation of endogenous antioxidants such as vitamins C and E and thiols as well as membrane cholesterol in isolated rat brain mitochondria were studied. Oxidation was induced by incubating the mitochondria at 37 degrees C with the free radical generators 2,2' azobis (2'-amidinopropane) dihydrochloride (ABAPH) and 2,2' azobis (2,4-dimethyl) valeronitrile (ABDVN) which undergo thermal decomposition to yield free radicals. An approximate order for the in vitro ease of oxidation was: ascorbate >> alpha-tocopherol > sulfhydryls >> cholesterol. However, small amounts of ascorbate were present in the mitochondria when alpha-tocopherol and sulfhydryl compounds were getting oxidized. This observation is different from those with more homogeneous biological substrates like blood plasma or serum. The order of oxidation of the various compounds is a function of not only the redox potentials but also the (a) concentrations of the oxidized and reduced species, (b) compartmentation of the compounds and (c) enzymatic and nonenzymatic systems for the repair or regeneration of the individual antioxidants. Even though ascorbate levels are quite low within mitochondria this nutrient may play a major role as a first line of defense against oxidative stress. The lipid-soluble ABDVN was much more potent in oxidizing membrane alpha-tocopherol and thiols than the water-soluble ABAPH. With both free radical generators the rate of oxidation of the antioxidants consisted of two phases. The initial phase, that is more rapid, may represent a pool of antioxidant that is involved in immediate antioxidant protection of the organelle with the slower compartment being responsible for replenishing the faster pool whenever needed.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7492952 TI - Membrane Ca2+ fluxes in rat hepatoma cells exposed to a supraphysiological concentration of asparagine. AB - It has been shown that supraphysiological concentrations of asparagine and hypoosmotic shock stimulate ornithine decarboxylase activity in cultured cancer cells by increasing the synthesis and the half-life of the enzyme protein. Since extracellular Ca2+ is essential for the action of asparagine and is also important for cell volume regulation in certain cell types, aspects of Ca2+ physiology in asparagine-treated H-35 rat hepatoma cells were investigated. The initial rate of influx of 45Ca increased from 0.25 to 1.04 nmol/min/mg protein immediately after exposure to 10 mM asparagine. With a one-minute lag the efflux rate also increased 2.2-fold over a five minute period. Asparagine did not cause a net-gain in cellular Ca2+ as measured by 45Ca equilibration, nor did it have any effect on the cytosolic free Ca2+ as measured by Fura-2 fluorescence spectroscopy and Fluo-3 fluorescence confocal microscopy. PMID- 7492953 TI - Increase of protooncogene c-myc mRNA levels by low density lipoprotein in vascular smooth muscle cells. AB - The effect of low density lipoprotein (LDL) on the intracellular mRNA concentration of the protooncogene c-myc was studied in freshly isolated bovine vascular smooth muscle cells and in the rat aortic smooth muscle cell line A7r5. Northern analysis showed that LDL increased the mRNA levels of c-myc in both cell lines, the stimulation being 2-fold after 2 h incubation at a concentration of 50 micrograms LDL-protein/ml. High density lipoprotein (HDL) had no effect on c-myc mRNA levels in A7r5 cells. These results demonstrate that LDL, but not HDL, increases intracellular concentrations of c-myc in two different aortic smooth muscle cell lines. PMID- 7492954 TI - Evidence that polymorphism of the angiotensin I converting enzyme gene may be related to idiopathic dilated cardiomyopathy in the Chinese population. AB - Angiotensin I-converting enzyme (ACE) is responsible for the production of angiotension II and the breakdown of kinins, leading to increased blood pressure (BP), induction of vascular smooth muscle cell proliferation, and the stimulation of myocardial-cell hypertrophy. A 287 bp insertion/deletion polymorphism in intron 16 of the ACE gene was examined by polymerase chain reaction in a cross sectional study of 35 patients with idiopathic dilated cardiomyopathy (IDC) and 35 patients with normally functioning hearts (NT). Compared with the deletion/deletion (D/D) frequency in the control population, the frequency of the deletion allele was 0.757 in IDC patients and 0.600 in NTs; the difference between observed alleles in all subjects in each group was significant (x2 = 3.96; P < 0.05). The data thus provide evidence in favor of an association between idiopathic dilated cardiomyopathy and a polymorphism at the ACE locus (17q23), thus implicating this locus, and possibly a genetic variant of ACE, itself, in human idiopathic dilated cardiomyopathy. PMID- 7492955 TI - Degradation of cystathionine gamma-lyase in rat liver lysosomes: effect of leupeptin treatment. AB - Highly purified lysosomes from the livers of normal and leupeptin-treated rats were subjected to immunoblot analysis, using antibodies against rat liver cystathionine gamma-lyase. Normal lysosomes showed small amount of single band at about 40 kDa, whereas the leupeptin-treated lysosomes showed large amounts of a major band at about 40 kDa and a minor band at about 35 kDa. These observations suggest that cystathionine gamma-lyase is sequestered into lysosomes at the original subunit molecular weight and is degraded in the lysosome by way of the sequential formation of an intermediate with distinct molecular weight. PMID- 7492956 TI - Polyamines exert regulatory control on mycobacterial transcription: a study using RNA polymerase from Mycobacterium phlei. AB - We have used purified RNA polymerase from Mycobacterium phlei to study the role of polyamines in mycobacterial transcription. Both initiation and elongation phases of the process were affected biphasically by polyamines. Interaction of polyamines with DNA template plays an important role in transcription modulation. Our studies emphasize that polyamines can exert a regulatory control on mycobacterial transcription. PMID- 7492957 TI - Purification and partial characterization of a lectin from Euphorbia neriifolia latex. AB - A lectin was purified from Euphorbia neriifolia latex to homogeneity by affinity chromatography on Sepharose 4B. The protein appears to be a dimer with approximate M(r) of 60,000 on gel filtration and showing a single band at M(r) 32,000 in SDS-PAGE, and contains 12.3% carbohydrate. It agglutinated trypsinized human and rabbit erythrocytes, but not sheep erythrocytes. However, sialidase treated sheep erythrocytes were agglutinated. The galactose and galactose containing sugars inhibited the heamagglutination with increased beta-anomeric specificity. The Euphorbia lectin possesses mitogenic activity with murine spleen lymphocytes but it does not inhibit protein synthesis in rabbit reticulocyte lysate. PMID- 7492958 TI - Sequences responsible for transcription termination of the mouse gastrin gene. AB - To identify the transcription termination elements in the mouse gastrin gene, we examined RNA transcripts after stable transfection of gastrin expression plasmids into the NIH3T3 cell line. The GT-repeat region at the 3'-flanking sequence of the mouse gastrin gene acted as a transcription terminator. When the GT-repeat unit was deleted from its site, the effect of termination disappeared. Further experiment, using serial deletion mutants, revealed that the 56-38 nucleotide upstream region from the GT-repeat unit also participated in transcription termination. We propose that the upstream region of the GT-repeat unit might be recognized as a pause site by the RNA polymerase II, and an abnormal DNA structure, derived from the GT-repeat unit, might function as a blockage of polymerase processivity. PMID- 7492959 TI - Status of membrane lipids and amino acid transport in morphological mutants of Candida albicans. AB - The phospholipid composition of various morphological mutants of Candida albicans revealed a complete absence of phosphatidylinositol (PI) from plasma membranes of those cells which completely lacked mycelial growth. No other phospholipid was found to be specific to morphogenesis. The plasma membrane fractions isolated from mutants were more rigid than its wild type as was evident from their unsaturation index and fluorescence polarization measurements. The enhanced membrane rigidity of mutant cells was noted regardless whether the cells could grow only as mycelia or in their budding forms. Although some amino acids are considered to affect the morphological transition of C. albicans, this was not reflected in the transport activities of L-proline, L-alanine, L-lysine and L glutamic acid. PMID- 7492960 TI - beta-Glucosidase families revealed by computer analysis of protein sequences. AB - This computational study is a summary of how cloned beta-glucosidase subfamilies are organized. Computations were carried out using General Computer Group, Inc. (GCG) package programs. Twenty-two beta-glucosidases belonging to either cellulolytic or non-cellulolytic organisms were identified. The multialignment of a whole beta-glucosidase family is shown. Two sub-families, A and B, were clearly seen to exist. Sub-family A is further subdivided into sub-families A1 and A2. A1 includes vegetal beta-glucosidases and A2 includes prokaryotic enzymes. Sub family B has three new sub-families, B1, B2, and B3. The enzymes in B2 are of yeast and/or fungi. Aspartic (D), glutamic (E) and histidine (H) residues, which are thought to be a part of the mechanism of the enzymatic hydrolysis are conserved. The well conserved amino acid sequences of the sub-family A are ITENGA; QUIEGA; HVD; and NEP. The well conserved amino acid sequences of the sub family B are: SDW; and YN(R,K)(V,L)N. PMID- 7492961 TI - Temperature-induced functional and structural transformations of the photosystem II oxygen-evolving complex in spinach subchloroplast preparations. AB - Heat inactivation of the process of O2 evolution, temperature-induced Mn2+ release and structural transitions revealed by differential scanning calorimetry (DSC) were studied in photosystem II (PSII) enriched subchloroplast fragments, granal thylakoids and the isolated oxygen-evolving pigment-lipoprotein complexes (OEC). It was found that the temperature of semi-inactivation of O2 evolution, which coincided with Mn2+ release, declined from 45 degrees C to 40 degrees C and 34 degrees C in this series of preparations, in accordance with the decreased structural stability of OEC. This was paralleled by a decrease in the content of light harvesting complex (LHC) and by an increase in the accessibility of OEC to hydrophilic electron acceptors. Thermoinactivation processes were accompanied by a two-fold decrease in PSII particle size on the EFs surface of membrane fragments. A "bi-core" oxygen-evolving complex model is proposed to account for these findings. PMID- 7492962 TI - Comparison between thymidylate synthase B of Bacillus subtilis ATCC6633 and 168. AB - Bacillus subtilis 168 possesses two thymidylate synthase genes: thyA and thyB, encoding for a thermolabile and a thermostable enzyme respectively. B. subtilis ATCC6633 also possesses two thy genes, both producing thermostable enzymes. A comparison of the thymidylate synthase B amino acid sequences from the two strains shows, among others, a Pro to Ala mutation which may affect the dUMP binding site. The apparent Km and Vmax values for dUMP and tetrahydrofolate were determined at a permissive temperature for both enzymes. The kinetic data showed a significant difference in the Km, but not in the Vmax, for dUMP between the two enzymes. The Km and Vmax for tetrahydrofolate were very similar. PMID- 7492963 TI - Production of melanin pigments by chemical and enzymatic oxidation of tetrahydroisoquinolines. AB - Tetrahydropapaveroline (THP) oxidation was studied in various experimental conditions by absorbance spectroscopy. THP was found to be easily oxidized by mushroom tyrosinase, giving rise to the formation of a chromophore (THP-chrome) with absorption maxima at 308 and 470 nm. The oxidation further proceeds leading to the formation of a melanin-like pigment. The use of periodate as oxidant at pH 7.4 allows the visualization of the THP-chrome, as well. Other tetrahydroisoquinolines bearing a catechol moiety, such as salsolinol, laudanosoline and apomorphine, have been found to be easily oxidized in the same conditions, giving rise to pigmented derivatives. The products of THP oxidation are able to copolymerize with dopa or opioid peptides in the presence of tyrosinase, generating mixed-type melanins. PMID- 7492964 TI - Molecular genetic characterization and prenatal diagnosis in a family with Hunter disease. AB - Hunter disease, Mucopolysaccharidosis type II, is an X-linked recessive lysosomal storage disorder caused by a deficiency in iduronate sulfatase activity. We studied at molecular level a Neapolitan family with the disease. We report, in patient, the delta 139 mutation on the third exon of the gene, on female family members, the DNA analysis that allowed to assess or exclude their carrier status and on fetal DNA from a pregnancy of patient's mother, a prenatal diagnosis that resulted negative. PMID- 7492965 TI - Binding of 241Am to carp cytosolic fractions in vitro. AB - Recent studies on the competition between Cd and radionuclides have been reported in marine species. The present investigation aims to study the binding of 241Am to freshwater fish cytosolic fraction before and after Cd-treatment. The main organs studied are the liver, kidneys and muscles. The distribution of americium in the liver cytosol of carp cadmium treated shows a preferential fixation on a 10 KDa fraction with high sulfhydril compounds. PMID- 7492966 TI - Cu/Zn-superoxide dismutase and glutathione peroxidase during aging. AB - During oxidative metabolism harmful reactive oxygen species (ROS) are generated. These species are neutralized by antioxidant enzymes. Firstly, superoxide dismutase (Sod) converts superoxide radicals (.O2-) to hydrogen peroxide (H2O2). Thereafter catalase (Cat) and glutathione peroxidase (Gpx) independently convert this to water. An imbalance in the ratio of Sod to Gpx and Cat results in the accumulation of H2O2 which may participate in the Fenton reaction, resulting in the formation of noxious hydroxyl radicals. These ROS are highly reactive and cause damage to macromolecules such as DNA, protein and lipids. We propose that it is the balance in the activity of the Sod to Gpx plus Cat ratio (Sod/(Gpx plus Cat)) that is an important determinant of cellular aging. This is based on our observation that an altered Cu/Zn-superoxide dismutase (Sod1)/(Gpx1 plus Cat) ratio exists in the brain of aging mice and that this correlates with increased lipid damage. Conversely, aging liver and kidney have an unaffected Sod1/(Gpx1 plus Cat) ratio and lipid damage is not increased with aging. We also examine the Sod1 to Gpx1 ratio in Down syndrome tissue and show that all organs have an altered ratio. This may contribute to the premature aging seen in these individuals. We show that binding of a p50/p65 complex to an NF-kappa B consensus sequence is enhanced by H2O2 treatment in NIH3T3 cells. Thus an altered Sod1/(Gpx1 plus Cat) ratio may also affect gene expression by altering the binding and/or availability of transcription factors to DNA. PMID- 7492967 TI - Novel use of limited primer extension in detecting mutations in human iduronate 2 sulfatase gene. AB - Hunter syndrome is characterized by a deficiency of iduronate 2-sulfatase. A large number of mutations in the gene have been reported. We describe here the development of a limited primer method for the identification of mutations. In the reaction mixture designed for the limited primer extension, one or two deoxynucleotides from the four necessary deoxynucleotides are added as "selected nucleotides" and another deoxynucleotide which is radiolabeled is added as the "limited nucleotide". The absence of one or two of the deoxynucleotides limits the length of primer elongation, and the low concentration of the "limited nucleotide" causes an "extension-delay" effect and results in a banding pattern upon electrophoresis of the products thus making it possible to distinguish mutant and normal alleles. We have studied three novel mutations in exon IX, 407delTT (TTT to T), 423insCC (CCC to CCCCC), and W502X (TGG to TAG) of the iduronate 2-sulfatase gene by the limited primer extension method. PMID- 7492968 TI - The molecular chaperone mhsp72 is partially associated with the inner mitochondrial membrane both in normal and heat stressed Spinacia oleracea. AB - The mitochondrial molecular chaperones mhsp70 and hsp60 in yeast have been shown to be essential components of the protein import and assembly machinery. Using the Western blotting technique and heterologous antibodies we have found that in spinach leaf mitochondria, a 72 kDa protein was an analogue of the mhsp70 and a 58 kDa protein, an analogue of the hsp60. Surprisingly, 49% of the spinach mhsp72 was associated with the membrane. The spinach hsp58 was entirely located in the matrix. The mhsp72 was loosely bound to the mitochondrial inner membrane. The heat shock treatment of the intact spinach plants resulted in a 2.4-fold induction of the mhsp72 and additionally facilitated association of this chaperone with the mitochondrial membrane. PMID- 7492969 TI - Evidence for autocrine growth inhibition of rat intestinal epithelial (RIE-1) cells by transforming growth factor type-beta. AB - The proliferation of sparse cultures of RIE-1 rat intestinal epithelial cells was potently inhibited by transforming growth factor type beta 1 (TGF-beta 1), with a half-maximal effect at 10-30 pg/ml. As judged by [3H]thymidine incorporation assays, this growth inhibitory action became apparent 4-6 h after addition of TGF beta 1 to the cells. RIE-1 cells express high-affinity (KD approximately 5.6 pM) receptors for 125I-TGF-beta 1. Affinity cross linking experiments labelled two major species of putative TGF-beta 1 receptors with M(r) values of 235,000 and 65,000. Medium conditioned by confluent cultures of RIE-1 cells contained latent TGF-beta-like activity that was activated at low pH. These findings support a model of autocrine growth inhibition of intestinal crypt cells by TGF-beta. PMID- 7492970 TI - Rat small intestine expresses primarily type VI gamma-glutamyl transpeptidase RNA. AB - To clarify differences between gamma-glutamyl transpeptidase (gamma GT) expression in the rat and the mouse small intestine, we cloned and sequenced 13 rat small intestine gamma GT cDNAs and compared them with those obtained from the mouse (Carter, et al. (1994) J. Biol. Chem. 269, 24581-24585). We found that all 13 were type VI, which in both species accounts for 80-90% of intestinal gamma GT. However, rat type VI gamma GT is substantially longer than mouse type VI RNA and includes at least 67 additional nucleotides at the 5' end that are non homologous with the 5' end of the mouse type VI RNA; further, these show no homology to another mouse exon (VII) that splices to type VI RNA. In addition, a 168 nucleotide stretch is present in the middle of rat RNA that is absent in mouse type VI RNA. PMID- 7492971 TI - Changes in cystathionine gamma-lyase in various regions of rat brain during development. AB - Developmental changes in the activities of cystathionine beta-synthase and cystathionine gamma-lyase were measured in six regions of rat brain. On day-1, no differences were observed in the activities of cystathionine beta-synthase and cystathionine gamma-lyase among these regions, the values being about 40 nmol/h/mg protein, and 3 nmol/h/mg protein, respectively. Cystathionine beta synthase activity increased gradually during development at almost the same rate in each region, reaching the adult level at week-4 (about 4-fold). Cystathionine gamma-lyase activity also increased during development, reaching adult level at week-2. But, the increase of enzyme activity in the cerebellum (about 1.8-fold) was clearly lower than that in the other regions (about 4-fold). Cystathionine gamma-lyase content in the various regions of week-3 rat brain estimated by immunoblotting was consistent with the enzyme activity, and the enzyme level in the cerebellum was lower than that in the other regions. Cystathionine content of cerebellum in week-3 increased rapidly during development, and was about five fold more than that on day-1. However, cystathionine content in the other regions did not change during development. These findings indicated that at least one reason of the high content of cystathionine in the 3 weeks rat cerebellum was due to the low level of cystathionine gamma-lyase. PMID- 7492972 TI - Activation of neuronal nitric oxide synthase by flavin adenine dinucleotide. AB - Bovine adrenals were found to contain a factor that activates neuronal nitric oxide synthase (NOS) and reduces the blood pressure when injected into the lateral cerebral ventricle (LCV). This factor showed chemical and functional characteristics similar to flavin adenine dinucleotide (FAD). Therefore, the effect of FAD on neuronal NOS activity was examined. FAD caused at least 2-fold stimulation of NOS partially purified from rat brain. This effect was not simply due to formation of the holoenzyme. Kinetic analyses showed that NOS exhibited negative cooperativity with L-arginine, its substrate, and FAD counteracted this effect. Furthermore, injection of FAD into the LCV reduced the blood pressure. These results suggest that FAD stimulates neuronal NOS by counteracting its negative cooperativity with L-arginine and also lowers the blood pressure by activating NOS. PMID- 7492973 TI - Potentiation of beta-adrenoceptor agonist mediated-lipolysis by cholesterol derived oxysterols. AB - Cholesterol-derived oxysterols such as cholestanol, cholestanone and coprostanone were able to potentiate epinephrine-induced lipolysis in intact rat adipocytes but not cholesterol. The relative potency of the oxysterols followed the sequence: cholestanone > or = coprostanone > cholestanol. Cholestanone was selected to study its mode of action on epinephrine-induced lipolysis. A sustained increase in the level of cAMP was observed in adipocytes incubated with both cholestanone and epinephrine compared to a transient peaking of cAMP in adipocytes incubated with epinephrine alone. Binding assays using [125I]cyanopindolol (beta-adrenergic receptor antagonist) showed that cholestanone could increase the binding affinity of [125I]-cyanopindolol to beta adrenergic receptors on rat adipocyte ghost membranes without affecting the total number of binding sites. The results suggest that cholestanone exerts its potentiation effect by facilitating the binding of beta-adrenergic agonist to its receptor. PMID- 7492974 TI - Determination of tissue endothelin levels. AB - A methodology for the quantitation of tissue endothelin levels has been developed. About 85% of authentic endothelin-1 added to the tissue extract was recovered. Using this protocol, the levels of endothelin in various rat tissues were determined. In Wistar-Kyoto rats, the kidney was found to have the highest level of endothelin, 1120 pg/gm wet weight, followed by the spleen and liver. The brain contained only half as much of endothelin when compared with the kidney. This method can be utilized to assess the pathological role of endothelin in cardiovascular or renal diseases. PMID- 7492975 TI - Use of nuclear magnetic resonance spectroscopy and selective C-labeling for pharmacokinetic research in man: detection of benzoic acid conversion to hippuric acid. AB - This paper demonstrates that the stable isotope tracer technique using NMR spectroscopy and the selective 13C labeling of protonated carbons can provide a relatively sensitive method to investigate pharmacokinetic problems in man. The urinary excreted [1,3,5-13C3]hippuric acid ([13C]HA) formed from orally administered [1,3,5-13C3] benzoic acid ([13C]BA) as a model substrate was successfully quantitated without any separation procedures by proton-decoupled 13C-NMR spectroscopy of 10-fold concentrated urine in a 10 min accumulation time. In spite of the low dosage (10mg BA), the C3,5 resonances of [13C]HA were detected with favorable signal-to-noise ratios to quantitate [13C]HA concentration. The administered [13C]BA was found to be quantitatively biotransformed to HA and excreted in urine within 4h. The lower limit of detection was estimated to be 50 nmol in an NMR tube, which was improved about one order of magnitude over that of BA labeled in the quaternary carbon (C7). The potential of an inverse detection experiment using heteronuclear multiple quantum coherence was also investigated in order to detect [13C]HA in urine, with a higher sensitivity. The inverse experiment improved the sensitivity by a factor of 2--3 over 13C?1H?-NMR, although the specificity of detection was relatively poor. PMID- 7492976 TI - Partial purification and characterization of an esterase acting on the anticancer pro-drugs, 7-ethylcamptothecin derivatives. AB - A hydrolytic enzyme which catalyzes hydrolysis of the ester-linkage of a series of 17-O-acyl derivatives of 7-ethylcamptothecin-21-(2-dimethylamino)ethylamide [acyl derivatives of 22E] was purified from rat liver and its properties were characterized. It hydrolyzed the ester-linkage of all 22E derivatives tested as well as p-nitrophenyl acetate at pH 8-9 but had no effect on 7-ethyl-10-[4 (piperidino)-1-piperidino] carbonyloxycamptothecin (CPT-11: irinotecan), unlike CPT-11 converting carboxylesterase, which was previously purified from rat serum [Tsuji T. et al., J. Pharmacobio-Dyn., 14, 341 (1991)]. The enzyme had no effect on either acetyl choline or butyrylcholine. It was inhibited by several organophosphorous compounds such as diisopropyl fluorophosphate (DFP), bis-(p nitrophenyl)phosphate and paraoxon, but was insensitive to inhibitors specific for choline esterases. These results indicate that this liver esterase is clearly distinct from choline esterase and serum CPT-11 converting enzyme and is able to convert pro-drugs, O-acyl derivatives of 22E, to an antitumor agent. PMID- 7492977 TI - Possible involvement of dihydrofructosazine in the DNA breaking activity of D glucosamine. AB - Dehydrochlorination of D-glucosamine (2-amino-2-deoxy-D-glucose) hydrochloride with an anion exchange resin made its DNA breaking activity in plasmid pBR322 much higher, especially in the presence of Cu2+. The sample of anion exchanger treated D-glucosamine hydrochloride, i.e., HCL-free D-glucosamine sample, showed an absorption maximum at 274 nm on the UV absorption spectrum in water as seen in the case of fructosazine [2,5-bis(D-arabino-tetrahydroxybutyl)pyrazine] one of the dimers of D-glucosamine. On a positive-ion fast atom bombardment (FAB) mass spectrum, the sample showed an ion peak at m/z 323 as a base peak, corresponding to dihydrofructosazine [2,5-bis(D-arabino-tetrahydroxybutyl) dihydropyrazine], which was a precursor of fructosazine, as well as those of D-glucosamine itself (m/z 180) and fructosazine (m/z 321). The DNA strand breaking activity of HCL free D-glucosamine sample was directly proportional to the peak intensity of m/z 323 ion, while the DNA breaking activity of fructosazine was much weaker than that of HCL-free D-glucosamine sample. 2,5-Dihydro-3,6-dimethylprazine and 2,3 dihydro-5,6-dimethylpyrazine having a dihydropyrazine skeleton the same as dihydrofructosazine showed the same extent of DNA strand breaking activity as did the HCL-free D-glucosamine sample. These results indicated that dihydrofructosazine produced during the dehydrochlorination is closely involved in the DNA breaking activity of HCL-free D-glucosamine sample. PMID- 7492978 TI - Evidence for accumulation of lipid hydroperoxides during the aging of human red blood cells in the circulation. AB - Young and old human red blood cells (RBC) were separated from freshly collected human blood by Percoll density gradient centrifugation. The amounts of lipid peroxidation products in the lipid fractions of these RBC were determined by high performance liquid chromatography-chemiluminescence, thiobarbituric acid and LPO 586 methods. The levels of phosphatidylcholine and phosphatidylethanolamine hydroperoxides in old RBC were higher than in young RBC. The products reflecting the advanced stage of lipid peroxidation, malonaldehyde (free and bound forms), 4 hydroxyalkenals (free and bound forms) and other aldehydes, were also higher in old RBC than in young RBC. The levels of these lipid peroxidation products increased when whole RBC were mildly oxidized with ferric ion. These results indicate that oxidative damage of RBC in the circulation takes place during the aging process, and lipid hydroperoxides and other lipid peroxidation products accumulate in old RBC. PMID- 7492980 TI - Localization and activity of adrenal carbonyl reductase during the estrous cycle. AB - Changes in carbonyl reductase (CR) activity towards seven substrates in adrenal glands of female rats were investigated during estrous cycle and the circadian patterns of adrenal CR activity towards seven substrates in male rats were examined. The localization of the enzyme protein in the adrenal gland was carried out by immunohistochemistry. In female rats, adrenal CR activity was lowest between 1200 and 2000 h on the day of proestrus, the phase in which a large, significant increase in ovarian steroids, gonadotropins and prolactin is observed. Immunoreactivity in the zona reticularis of the adrenal cortex to anti adrenal CR, CR-K2, antibody on the day of proestrus was faint or negative compared with that on other days of the estrous cycle. In male rats, no major changes in adrenal CR activity were observed compared with females, although these were slight changes in all the enzyme activities. These results suggest that adrenal CR may be negatively regulated by the pituitary-adrenal axis in female rats and, in particular, on the day of proestrus. PMID- 7492979 TI - Effect of a high alpha-linolenate and high linoleate diet on membrane-associated enzyme activities in rat brain--modulation of Na+, K+- ATPase activity at suboptimal concentrations of ATP. AB - Semi-purified diets supplemented with either a high alpha-linolenate (n - 3) (perilla) oil or a high linoleate (n - 6) (safflower) oil were fed to rats through two generations. Rats fed safflower oil showed a decrease in docosahexaenoic acid (n - 3) and a compensatory increase in docosapentaenoic acid (n - 6) in all the brain regions and organelles examined, when compared with rats fed perilla oil. As reported previously, the safflower oil-fed rats exhibited inferior learning ability compared with the perilla oil-fed rats (N. Yamamoto et al., J. Lipid Res. 28, 144 (1987)). Using brains of rats in these dietary groups, the activities of several enzymes, Na+ , K+-ATPase, Ca2+-ATPase, 5'-nucleotidase, 2',3'-cyclic nucleotide phosphodiesterase, acetylcholinesterase, and choline acetyltransferase in membranes, were compared. The 5'-nucleotidase activity in cortex and hippocampus, and the Na+, K+-ATPase activity in myelin decreased slightly but significantly in the safflower oil group. None of the other membrane associated enzyme activities in all the brain regions and organelles examined was affected significantly by the dietary fatty acids under optimal assay conditions in vitro. However, in the safflower oil group, the Na+, K+-ATPase activity of synaptosomes at a suboptimal concentration of ATP was 78% that in the perilla oil group. These results suggest that relatively large changes in the proportions of n - 3 and n - 6 polyunsaturated fatty acids in brain membranes caused by dietary manipulation do not provoke significant alterations in most membrane-bound enzyme activities. However, a small but significant change in Na+, K+-ATPase activity at a suboptimal concentration of ATP may be implicated in the altered learning behavior reported earlier. PMID- 7492981 TI - Bioavailability of calcium from calcium carbonate, DL-calcium lactate, L-calcium lactate and powdered oyster shell calcium in vitamin D-deficient or -replete rats. AB - The bioavailability of calcium from various calcium sources in humans and animals has been the subject of investigation for many years and there is considerable controversy as to the relative bioavailability of different calcium salts. Most of the studies have used a calcium balance technique which has numerous problems in terms of performance and interpretation. Using a method for evaluating the efficacy of calcium from calcium salts used for plasma calcium metabolism and bone mineralization, we examined the bioavailability of calcium from four commercially available calcium salts, namely calcium carbonate, DL-calcium lactate, L-calcium lactate and powdered oyster shell-calcium in vitamin D deficient or -replete rats. Among the calcium salts, the differences in bioavailability were small and not statistically significant as tested by analysis of variance in both groups of rats. Thus, we conclude that calcium is utilized to the same extent from calcium carbonate, DL-calcium lactate, L-calcium lactate and powered oyster shell-calcium in both vitamin D-deficient and -replete rats. PMID- 7492982 TI - Antiallergic constituents from oolong tea stem. AB - The antiallergic constituents of oolong tea stem were examined. The stem extracts inhibited the 48 h homologous passive cutaneous anaphylaxis (PCA) reactions or rats in a dose-dependent manner and showed the same extent of inhibitory activity as ketotifen. All antiallergic constituents from the stem were concentrated into chloroform and ethyl acetate fractions, when extracted by various solvents. These fractions were treated with polyvinylpolypyrrolidone (PVPP), which resulted in the elimination of antiallergic activity in the ethyl acetate fraction, suggesting that one of the antiallergic constituents may be tea catechins. Then, six kinds of catechins, (-)-epigallocatechin gallate (EGCG), (-)-epigallocatechin (EGC), (-)-epicatechin gallate (ECG), -epicatechin (EC), (+)-catechin (C) and (-) gallocatechin gallate (GCG), were isolated from the ethyl acetate fraction, and the inhibitory activity of these catechins on histamine release from rat peritoneal mast cells passively sensitized with anti-egg albumin (EA) IgE antibody was investigated. Among these catechins, significant inhibitory activity was observed in all the catechins except for EC. In addition, the inhibitory activity of GCG was greater than that of EGCG, which is well known to be an antiallergic constituent in tea. These results suggest that GCG may be a novel antiallergic constituent among tea catechins, and also the most potent. PMID- 7492983 TI - Role of angiotensin II in the transforming growth factor-beta 1 expression of rat kidney in anti-glomerular basement membrane antiserum-induced glomerulonephritis. AB - Induction of acute nephritis in the rat by injecting anti-glomerular basement membrane (GBM) antiserum is accompanied by a transient increase in angiotensin II generation in blood circulation within the first 24 h and a subsequent elevation of transforming growth factor-beta 1 (TGF-beta 1) mRNA levels in kidney cortex with a peak at days 7-8. Studies were carried out to determine whether the increased generation of angiotensin II plays a role in the elevation of TGF-beta 1 mRNA. Elevation of TGF-beta 1 mRNA levels 7 d after injection of antiserum was significantly inhibited by a successive daily administration of TCV-116, angiotensin II type 1 receptor antagonist, at 1 mg/kg/d from days 0 to 2 or from days 0 to 6, while it was not influenced by a single administration of this dose on day 0. In addition, angiotensin II infusion or 24 h at a rate of 50 ng/min did not alter the level of TGF-beta1 mRNA which was measured 6 d after the infusion. These results suggest that the anti-GBM antiserum-induced increase in TGF-beta 1 expression in the kidney is not responsible for angiotensin II generated in the blood circulation during the early phase of acute nephritis, but iis probably mediated by angiotensin II generated locally in the kidney. PMID- 7492984 TI - Depolarizing neuromuscular blocking action of coryneine derived from aconite root in isolated mouse phrenic nerve-diaphragm muscles. AB - The mode of the neuromuscular blocking action of coryneine (a quaternary ammonium derivative of dopamine) derived from aconite root was investigated in isolated phrenic nerve-diaphragm muscles and denervated diaphragm muscles of mice. Coryneine (20-150 microM) blocked the nerve-evoked twitch response without affecting the contraction evoked by electrical stimulation of the muscle. The blocking effect was reversed by neostigmine, a cholinesterase inhibitor. The electrical charge-response curve on depolarization produced by iontophoretically applied acetylcholine (ACh) at the endplate regions in normal muscles was shifted to the right on decreasing the maximal response by 40 microM coryneine. The double-reciprocal plot revealed that coryneine reduced the apparent affinity of ACh for its receptor on decreasing the maximal response. Coryneine (20 microM 2mM) itself depolarized the endplate membrane and this effect was reversibly suppressed by 1 and 5 microM pancuronium. Coryneine 30 microM-10mM) produced contractions of denervated muscles in a concentration-dependent manner and the effects were reduced by 70nM pancuronium. These results indicate that coryneine is a depolarizing agent and a mixed-type competitive and noncompetitive neuromuscular blocker. PMID- 7492985 TI - 3,4,5-Trimethoxyphenylacetaldehyde, an intermediate metabolite of mescaline, is a substrate for microsomal aldehyde oxygenase in the mouse liver. AB - 3,4,5-Trimethoxyphenylacetaldehyde, an intermediate metabolite of mescaline, was oxidized to 3,4,5-trimethoxyphenylacetic acid by mouse hepatic microsomes. The reaction was NADPH-dependent, and inhibited by SKF 525-A, metyrapone and disulfiram. A P450 isozyme in mouse hepatic microsomes, P450 MUT-2 (CYP2C29), catalyzed the reaction (0.96 nmol/min/nmol P450) in which NADPH and NADPH cytochrome c reductase were essential for the catalytic activity. The reaction was confirmed to be an oxygenation since molecular oxygen was incorporated into the carboxylic acid metabolite formed under oxygen-18 gas by GC-MS analysis. By addition of antibody against CYP2C29 to the microsomes (3.2 mg/mg microsomal protein) the MALDO activity was inhibited by 35% of the control value with preimmune serum, suggesting that CYP2C29 or an immunologically-related isozyme(s) plays a major role in the NADPH-dependent oxidation of 3,4,5 trimethoxyphenylacetaldehyde to 3,4,5-trimethoxyphenylacetic acid by mouse hepatic microsomes. Pharmacological experiments on mescaline and its deaminated metabolites using mice indicated that the metabolites were much less active or were inactive in cataleptogenic effect and pentobarbital-induced sleep prolongation as compared with the parent compound. PMID- 7492987 TI - Effect of cytochrome P450 inducers on liver microsomal metabolism of tetrachlorobiphenyls in rats, guinea pigs and hamsters. AB - Effect of cytochrome P450 (P450) inducers on liver microsomal metabolism of 3,4,3',4'-, 3,5,3',5'- and 2,5,2', 5'-tetrachlorobiphenyl (TCB) was studied using male Wistar rats, male Hartley guinea pigs and male Golden syrian hamsters. In metabolism of 3,4,3'4'- and 3,5,3',5'-TCB, liver microsomes from 3 methylcholanthrene (MC)- or 3,4,5,3',4'-pentachlorobiphenyl (PenCB)-treated hamsters showed hydroxylase activities for both TCB isomers, although the activities were much less than those of rats. In contrast, liver microsomes form untreated and phenobarbital (PB)-, MC- or PenCB-treated guinea pigs showed no hydroxylase activity. In 2,5,2',5'-TCB metabolism, 3-hydroxylase activity was observed in untreated guinea pigs and hamsters, but not in untreated rats. The activity pigs was induced by PB treatment in all three species, at rates of 324, 19 and 20 pmol/min/mg protein in rats, guinea and hamsters, respectively. This activity was not enhanced by treatment with either MC or PenCB. Only in hamsters was 4-hydroxylated metabolite formed in all microsomes used in addition to the 3 hydroxylated one, and the formation was accelerated 2.0-, 2.7- and 4.8-fold by treatment with PB, MC and PenCB, respectively. These results suggest that different P450 isoforms in hamster liver microsomes are involved in 3- and 4 hydroxylation of 2,5,2',5'-TCB. Thus, there are species differences in the basal ability to hydroxylate TCB isomers, and in the extent of effect of P450 inducers on the metabolism of these isomers among the three species. PMID- 7492986 TI - Effects of losartan, an angiotensin II antagonist, on the development of cardiac hypertrophy due to volume overload. AB - To investigate the contribution of a cardiac renin-angiotensin system to cardiac hypertrophy due to volume overload, the effects of losartan, a non-peptide angiotensin (Ang) II type 1 (AT1) receptor antagonist on left ventricular hypertrophy (LVH) was studied. LVH was produced in male Wistar rats by volume overload secondary to aortic insufficiency (AI). Losartan (10 mg/kg/d) was orally administered for 2 weeks after surgery to both AI and sham-operated (control) rats. Two weeks after surgery, aortic pulse pressure and left ventricular (LV) weight were markedly increased in the AI rats as compared with the control group, whereas cardiac angiotensin converting enzyme (ACE) activity remained unchanged. The effects of the chronic administration of losartan an AT1 receptors were verified by the blockade of Ang II pressor response. Losartan treatment produced a significant reduction in LVH in AI rats without affecting the systolic blood pressure. In separate groups of rats, to elucidate the mechanisms of the attenuation of LVH by treatment with losartan, we determined plasma and LV immunoreactive Ang II content and plasma renin activity (PRA). LV Ang II content increased in AI rats, while plasma Ang II content, PRA and II concentration were increased by the treatment. There was a significant positive correlation between LV weight and LV Ang II content. These results suggest that cardiac Ang II, rather than circulating Ang II, plays an important role in the LVH due to volume overload via the AT1 receptor. PMID- 7492988 TI - Direct effect of 12(S)-hydroxyeicosatetraenoic acid on rat crystalline lens is perturbation of lens normalcy. AB - The direct effect of 12(S)-hydroxyeicosatetraenoic acid [12(S)-HETE] on rat crystalline lens was investigated in this study. 12(S)-HETE lowered the glutathione (GSH) level and GSH reductase activity in the lens, while accelerating aggregation and insolubilization of lens proteins and production of thiobarbituric acid-reactive substances. The study also indicated that 12(S)-HETE insolubilized alpha-crystalline and induced opacification of the lens when the lens was incubated with 12(S)-HETE. From the results, we presumed that 12(S)-HETE may be oxidized or peroxidized easily and automatically in the air. The substances derived from 12(S)-HETE by oxidation or peroxidation may give the action disordering lens normalcy and induced cataract formation. Thus, the direct effect of 12(S)-HETE may of no benefit to the crystalline lens. PMID- 7492990 TI - Cytotoxic activity of bryonolic acid isolated from transformed hairy roots of Trichosanthes kirilowii var. japonica. AB - Bryonolic acid was isolated in high yield from transformed hairy root cultures of Trichosanthes kirilowii var. Japonica. Bryonolic acid exhibited cytotoxic activity to various tumor cells in vitro, independent of cell type. Normal cells such as rat hepatocytes are less sensitive to bryonolic acid. The appearance of a DNA ladder was detected in the bryonolic acid-treated HL-60RG cells, indicating that cell death triggered by bryonolic acid is due to apoptosis. PMID- 7492989 TI - Mechanism on insulin-like action of vanadyl sulfate: studies on interaction between rat adipocytes and vanadium compounds. AB - When rats with streptozotocin (STZ)-induced diabetes were given a daily intraperitoneal (i.p.) injection of VOSO4 (+4 oxidation state of vanadium), their serum glucose dropped from hyperglycemic level to normal level within 2d and serum free fatty acid (FFA) level also dropped to normal level. Vanadium was incorporated in most organs as well as in the adipose tissues, as detected by neutron activation analysis (NAA). The mechanism for the insulin-like action vanadium in terms of FFA release from isolated rat adipocytes was investigated: (1) Vanadyl (IV) and vanadic (III) ions normalize the FFA release in the adipocytes treated with epinephrine; (2) vanadate (V) ion treated with ascorbic acid, cysteine or glucose is effective in normalizing the FFA release but vanadate ion alone has no effect on FFA release; (3) vanadyl ion is incorporated into the adipocytes, while vanadate ion is not, as indicated by ESR spectroscopy; and (4) vanadyl ion can act on the glucose transporter, as indicated by experiments using cytochalasin B which is an inhibitor of this transporter. From these results, the normalization of both serum glucose and FFA levels by vanadyl ion was concluded to be due to the incorporation of vanadyl ion into the adipocytes, in which the metal ion acts on the glucose transporter and induces both the promotion of glucose uptake and the decrease of FFA release form peripheral adipocytes. The vanadyl state was suggested to be a possible pharmacologically active form of vanadium allowing the insulin-like action.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7492991 TI - Inhibitory effect of rikkunshi-to, a traditional Chinese herbal prescription, on tumor promotion in two-stage carcinogenesis in mouse skin. AB - We report the inhibitory effect of topical application of extracts of a traditional Chinese herbal prescription on 12-O-tetradecanoylphorbol-13-acetate (TPA)-induced inflammation in mice. Methanol and water extracts obtained from 22 traditional Chinese herbal prescriptions were assayed and their inhibition ratios calculated. In general, the methanol extracts produced more effective inhibition than the water extracts. Of the various traditional Chinese herbal prescriptions, the methanol extract of Rikkunshi-to was more effective than other prescriptions as far as inhibition of TPA-induced inflammation was concerned. Hoelen, Glycyrrhizae Radix, Atractylodis Rhizoma, components of Rikkunshi-to markedly inhibited the inflammatory activity involved by TPA in mice. Furthermore, topical application of the methanol extract of Rikkunshi-to markedly inhibited TPA induced tumor promotion in two-stage carcinogenesis in mouse skin. PMID- 7492993 TI - Syntheses of novel galactosyl ligands for liposomes and the influence of the spacer on accumulation in the rat liver. AB - We modified the surface of liposomes with galactosyl ligands. At first we determined whether or not the galactosyl moiety was exposed on the liposomes. We then investigated the effect of the ligands on the hepatic accumulation of liposomes in rats. We introduced an oligoethylene glycol moiety as a spacer. Among the various ligands tested, those with a tri- or tetraethylene glycol moiety as a spacer caused the greatest accumulation of liposomes in the liver. Liposomes bearing ligands with a tri- or tetraethylene glycol moiety as a spacer, were aggregated by Ricinus communis agglutinin. On the other hand, those modified with ligands with a mono- or diethylene glycol spacer did not clearly agglutinate. These results show the importance of a spacer between the homing device and the ligand anchor. PMID- 7492992 TI - Effects of dose, pH and osmolarity on intranasal absorption of recombinant human erythropoietin in rats. AB - The effects of dose, pH and osmolarity on the intranasal absorption of a recombinant human erythropoietin (rEPO) solution were studied in male Wistar rats. The intranasal administration of rEPO was evaluated by measuring percentage circulating reticulocytes of red blood cells on a stained blood smear (smear method), and also by measuring residual circulating reticulocyte counts using a microcell counter (sysmex method). Both results suggest that rEPO solution was absorbed through the nasal mucosa of rats without enhancers after a single intranasal administration. The pharmacological availabilities of rEPO after intranasal administration compared with intravenous administration were about 7% and 4%, when estimated by smear method and sysmex method, respectively. The pharmacological activity was enhanced in low pH and hypotonic mannitol solution. PMID- 7492994 TI - Influence of anesthetic regimens on the intestinal absorption of 5-fluorouracil in rats. AB - We investigated the influence of anesthetic regimens on the intestinal absorption of 5-fluorouracil (5-FU), which is known to be absorbed by concurrent Na(+) dependent, carrier-mediated transport and passive transport, in single-pass perfusion experiments in rats. Compared with the absorption in unanesthetized rats, the regular dose of urethane (1.13g/kg) reduced the maximum transport rate (Jmax), the Michaelis constant (Km) and the membrane permeability coefficient of passive transport (P m,d); a low dose of urethane (0.7g/kg) reduced Jmax and Kmax, but did not affect Pm,d; pentobarbital sodium (50 mg/kg) increased Jmax without affecting Km, and reduced Pm,d. The reductions in Jmax and Km were comparable for the regular and low doses of urethane. Thus, urethane and pentobarbital, which have been most commonly used in laboratory animal experiments, exerted qualitatively different effects on the carrier-mediated transport of 5-FU, although they similarly inhibited the passive transport. For urethane, the effect on the passive transport was avoided by reducing the dose, but the effect on the carrier-mediated transport was not. This influence of anesthetic regimens on intestinal drug absorption may not be easily scaled for normalizing absorption data. When compiling them for such purposes as establishing in situ-in vivo quantitative correlation, the absorption data in perfusion (in situ) should be categorized on the basis of anesthetic regimens, to avoid ending up with poor outcomes. We also examined the effect of urethane on the exsorption of Na+ in the intestinal loop where Na+-free buffer was introduced, and found a minimal effect.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7492995 TI - Noninvasive measurement of cerebral blood flow and glucose metabolic rate in the rat with high-resolution animal positron emission tomography (PET): a novel in vivo approach for assessing drug action in the brains of small animals. AB - The cerebral metabolic rate of glucose (CMRglc) and cerebral blood flow (CBF) in rats were estimated noninvasively with a high-resolution animal positron emission tomography (PET) system that we recently developed. Qualifications of CMRglc and CBF were attempted with radiotracers, 18F-2-fluoro-2-deoxyglucose and 15O-water, respectively. From the data obtained with PET, the CMRglc and CBF values under pretreatment conditions were respectively calculated as 51 +/- 14 mumol/min/100 g and 61 +/- 7 ml/min/100 g under pentobarbital-anesthesia. These values were in close agreement with published data obtained with the autoradiographic technique. Moreover, an increase of 69% (mean) in CBF was detected after i.v. acetazolamide administration. The results indicate that this high-resolution animal PET scanner holds great potential for the assessment of drug-related metabolic and circulatory effects in intact and experimental animals. PMID- 7492996 TI - Characterization of mitogenic substances in the hot water extracts of bupleuri radix. AB - Bupleuri Radix is a commonly used medicinal plant in Kampo medicine, and its hot water extracts show mitogenic activity to murine lymphocytes. In this paper the mitogenic substances in the hot water extracts of Bupleuri Radix (Bup-HWE) were fractionated and characterized physicochemically and immunologically. Most of these substances were recovered from mol. wt of more than 200 kDA fraction (fr. C 13). Separation of fr. C-13 by phenol-water fractionation method gave water soluble and phenol soluble mitogenic substances. These substances showed the activity even in C3H/HeJ mice, and polymyxin B or lysozyme treatment did not abrogate the activity, suggesting that the active substances are not related to bacterial lipopolysaccharide. Treatment of the mitogenic substances recovered from the phenol layer with NaCLO2, a polyphenol degrading chemical, significantly reduced the activity, but pronase and pectinase treatments were not effective. The mitogenic substances in the water layer were active even after NaCLO2 treatment. These findings suggested that the mitogenic substances of Bup-HWE are large molecular weight polyphenolic compounds and polysaccharide. The mitogenic substances are suggested to be B cell mitogens. PMID- 7492997 TI - Stabilization of Octastatin, a somatostatin analogue. Preparation of freeze-dried products for parenteral injection. AB - Octastatin (RC-160, vapreotide INN) is a somatostatin analogue being developed for use in oncological, enterologic and neuroendocrine applications. The pharmaceutical form is a freeze-dried preparation for parenteral injection use. Three dosage forms containing 0.5, 1.5 and 15 mg of vapreotide base have been investigated. Various freeze-drying conditions, stabilizing agents, membranes for sterile filtration and heating procedures have been examined. The formulation with glutamic acid-sodium glutamate buffer as a stabilizing agent, the type of membrane for filtration and the freeze-drying procedure have been found appropriate for subsequent industrial production. No evident degradation was observed either after manufacturing, or after a three-week accelerated stability study at 50 degrees C and 70% relative humidity. PMID- 7492998 TI - Modulation of the antitumor effect and tissue distribution of highly branched (1- >3)-beta-D-glucan, SSG, by carrageenan. AB - The action of carrageenan (CAR), a representative blocking reagent for phagocytes, on the antitumor effect and tissue distribution of highly branched (1 ->3-beta-D-glucan, SSG, was examined. CAR inhibited the antitumor effect of intraperitoneally administered SSG only when applied before inoculation of the tumor, and had little effect when applied after tumor inoculation. A similar result was observed when SSG was administered intralesionally. In contrast, CAR had considerable effect on tissue distribution of i.p. SSG. The differences with respect to the results in normal mice were: 1) the distribution of SSG from the peritoneal cavity to the rest of the body was inhibited, 2) large numbers of peritoneal exudate cells (PEC) were produced and a relatively high concentration of 3H-SSG was found in the PEC fraction 48h after administration of 3H-SSG, 3) one week after administration, 3H-SSG was distributed to throughout the body but the amount of 3H-SSG distributed was lower than in normal mice, 4) a significant amount of 3H-SSG was recovered from ligaments (containing omental milky spots, peritoneum, mesentery and associated fat) in which negligible amounts were found in normal mice. The results suggest that the inhibition of the antitumor effect of SSG by CAR probably results from the prevention of the natural resistance of mice which is related to phagocytic function, and that the distribution of SSG to throughout the body is significantly modulated by CAR. PMID- 7493000 TI - Effects of 6-(10-hydroxydecyl)-2,3-dimethoxy-5-methyl-1,4-benzoquinone (idebenone) and related benzoquinones on porcine pancreas phospholipase A2 activity. AB - 6-(10-Hydroxydecyl)-2,3-dimethoxy-5-methyl-1,4-benzoquinone (idebenone) has been found to have a membrane-stabilizing activity in studies using lysosomes. Using dimyristoyl L-alpha-phosphatidylcholine (DMPC) as a substrate, the effects of idebenone and related benzoquinones on phospholipid digestion by phospholipase A2 (PLA2) was investigated. Free myristic acid, released from DMPC upon PLA2 treatment, was anthrylmethylated with 9-anthryldiazomethane (ADAM) and determined by reversed-phase HPLC. Idebenone and a related benzoquinone, 2,3-dimethoxy-5 methyl-6-(10-morpholinodecyl)-1, 4-benzquinone hydrochloride (QS-10.Mor), inhibited the hydrolysis of the substrate with PLA2 in a dose-dependent manner. It is suggested that the effect of idebenone on PLA2, in addition to its antioxidant activity lipid peroxidation, can be attributed to membrane stabilizing activity. PMID- 7492999 TI - Manipulation of the lighting schedule can modify the pharmacological effects of theophylline in chick embryos. AB - The effect of the lighting schedule on the pharmacological action of theophylline was studied in chick embryos. Fertile eggs of White Leghorns were incubated and investigated, on two occasions, under constant light conditions or under constant dark conditions. A single injection of theophylline 2.14, 4.29, 8.57 and 17.14 mg/egg into the air sac of fertile eggs was carried out on the 16th day of incubation. Electrocardiograms (ECGs) were recorded 0 to 60 min after drug injection. After drug injection, heart rate increased under constant light conditions, but decreased under dark conditions. In addition, arrhythmia was produced by theophylline, 17.14 mg/egg, under constant dark conditions. These results indicate that the manipulation of the lighting schedule may have a marked influence on the pharmacological effects of theophylline in chick embryos. PMID- 7493002 TI - Hypoglycemic effect of the rhizomes of ophiopogonis tuber in normal and diabetic mice. AB - The hypoglycemic effect of the rhizomes of Ophiopogonis Tuber (Liliaceae) was investigated in normal and streptozotocin-induced diabetic mice. The n-butanol extract of rhizomes of Ophiopogonis Tuber (BM) (100 mg/kg) reduced the blood glucose of normal mice from 201 +/- 13 to 151 +/- 7 mg/100 ml 4h after intraperitoneal administration (p < 0.054), and also significantly lowered the blood glucose of streptozotocin-induced diabetic mice from 590 +/- 28 to 470 +/- 37 mg/100 ml under similar conditions (p < 0.05). BM also tended to suppress epinephrine-induced hyperglycemia in mice. We concluded that the hypoglycemic effect of BM does not alter the insulin concentration. PMID- 7493001 TI - Conformational analysis of 19-oxygenated steroids with a 4-ene or 2,4-diene structure, potential intermediates of aromatase reaction, with semiempirical molecular orbital PM3 calculations. AB - Conformational analysis of potent competitive inhibitors of aromatase, androst-4 enes 5, as well as 2,4-diene steroids 3, 4, and 6 was carried out, using theoretical calculations, to determine the stereochemistry of their aromatase catalyzed oxygenation. In the steroids examined, both the 19-alcohols and the 19 aldehydes favor the above- A ring conformation among the possible three in each. The results suggest that the 3-deoxy steroid 5a as well as the 2,4-diene steroids 4a and 6a would be oxygenated at C-19 by aromatase through the same stereomechanism as that involved in the androstenedione aromatization. PMID- 7493003 TI - Ca2+ dependency of diadenosine 5',5"'-p1,p4-tetraphosphate degradation in dog plasma. AB - Diadenosine 5',5"'-p1,p4-tetraphosphate (Ap4A) degradation in dog plasma was inhibited by the addition of EDTA. The Ap4A degrading activity seemed to be recovered by the addition of metal ions, Ca2+, Co2+, Cu2+, Mg2+, Mn2+ or Zn2+ to plasma in which the activity was lost by the addition of EDTA. Especially, the degradation was accelerated by the addition of Ca2+ to the plasma, and the degradation rate was dependent on the concentration of Ca2+. The results of this study suggested that the enzymatic degradation of Ap4A in dog plasma was modulated by the concentration of Ca2+. PMID- 7493004 TI - Accumulation of lauric acid in skin as an enhancer for the percutaneous absorption of thiamine disulfide. AB - The permeation and accumulation of lauric acid (12:0) in rat skin as an enhancer for the percutaneous absorption of thiamine disulfide (TDS), using propylene glycol as a vehicle, were determined in vitro. 12:0 barely permeated through skin, and accumulated in skin. The accumulated amount increased increasing concentrations in the vehicle (1-20%). The proportion of the accumulation to total dose was highest at a dose of 5%. A positive relationship was found between the accumulated amount of TDS and 12:0. It was suggested that the penetration of 12:0 into skin contributes to the enhanced partition of TDS to skin. PMID- 7493006 TI - Sexual orientation revolution. PMID- 7493007 TI - How do you compute a lod score? PMID- 7493005 TI - Does bacitracin have an absorption-enhancing effect in the intestine? AB - We studied the absorption enhancement effects of three types of protease inhibitors, aprotinin, bacitracin and soybean trypsin inhibitor, in the rat intestine. Of these protease inhibitors, bacitracin enhanced the absorption of FD 4 and phenol red from the rat small and large intestine without mucosal toxicity. Thus, it was suggested that bacitracin has not only a protease-inhibitory but also an absorption-enhancing capability. PMID- 7493008 TI - A DNA helicase in full Bloom. PMID- 7493009 TI - Calcium sensing comes full circle. PMID- 7493010 TI - Language replacement in Scandinavia. PMID- 7493012 TI - Toulouse-Lautrec's diagnosis. PMID- 7493011 TI - Microsatellite 'evolution': directionality or bias? PMID- 7493013 TI - Informed consent and BRCA1 testing. PMID- 7493014 TI - DNA statistics in the Simpson matter. AB - On 3 October 1995, O.J. Simpson was acquitted of two murders in spite of very strong DNA evidence linking his blood to the crime. Although numerical statements describing the strength of this evidence were made, the DNA profiles included so many loci that the need for presenting numbers in this case, and in others using similarly high numbers of loci, is probably unnecessary. If numbers are to be presented, however, they should be given in the form of likelihood ratios. One thing the verdict in the Simpson case makes clear is that it is essential that the integrity of DNA evidence (with regard to collection, potential contamination or tampering) be beyond doubt. PMID- 7493015 TI - The mouse Snell's waltzer deafness gene encodes an unconventional myosin required for structural integrity of inner ear hair cells. AB - The mouse represents an excellent model system for the study of genetic deafness in humans. Many mouse deafness mutants have been identified and the anatomy of the mouse and human ear is similar. Here we report the use of a positional cloning approach to identify the gene encoded by the mouse recessive deafness mutation, Snell's waltzer (sv). We show that sv encodes an unconventional myosin heavy chain, myosin VI, which is expressed within the sensory hair cells of the inner ear, and appears to be required for maintaining their structural integrity. The requirement for myosin VI in hearing makes this gene an excellent candidate for a human deafness disorder. PMID- 7493016 TI - Dilated cardiomyopathy and neonatal lethality in mutant mice lacking manganese superoxide dismutase. AB - The Sod2 gene for Mn-superoxide dismutase (MnSOD), an intramitochondrial free radical scavenging enzyme that is the first line of defense against superoxide produced as a byproduct of oxidative phosphorylation, was inactivated by homologous recombination. Homozygous mutant mice die within the first 10 days of life with a dilated cardiomyopathy, accumulation of lipid in liver and skeletal muscle, and metabolic acidosis. Cytochemical analysis revealed a severe reduction in succinate dehydrogenase (complex II) and aconitase (a TCA cycle enzyme) activities in the heart and, to a lesser extent, in other organs. These findings indicate that MnSOD is required for normal biological function of tissues by maintaining the integrity of mitochondrial enzymes susceptible to direct inactivation by superoxide. PMID- 7493017 TI - Insensitivity to anti-mullerian hormone due to a mutation in the human anti mullerian hormone receptor. AB - Anti-Mullerian hormone (AMH) and its receptor are involved in the regression of Mullerian ducts in male fetuses. We have now cloned and mapped the human AMH receptor gene and provide genetic proof that it is required for AMH signalling, by identifying a mutation in the AMH receptor in a patient with persistent Mullerian duct syndrome. The mutation destroys the invariant dinucleotide at the 5' end of the second intron, generating two abnormal mRNAs, one missing the second exon, required for ligand binding, and the other incorporating the first 12 bases of the second intron. The similar phenotypes observed in AMH-deficient and AMH receptor-deficient individuals indicate that the AMH signalling machinery is remarkably simple, consisting of one ligand and one type II receptor. PMID- 7493018 TI - A mouse model of human familial hypocalciuric hypercalcemia and neonatal severe hyperparathyroidism. AB - Mice lacking the calcium-sensing receptor (Casr) were created to examine the receptor's role in calcium homeostasis and to elucidate the mechanism by which inherited human Casr gene defects cause diseases. Casr+/- mice, analogous to humans with familial hypocalciuric hypercalcemia, had benign and modest elevations of serum calcium, magnesium and parathyroid hormone levels as well as hypocalciuria. In contrast, Casr-/- mice, like humans with neonatal severe hyperparathyroidism, had markedly elevated serum calcium and parathyroid hormone levels, parathyroid hyperplasia, bone abnormalities, retarded growth and premature death. Our findings suggest that Casr mutations cause these human disorders by reducing the number of functional receptor molecules on the cell surface. PMID- 7493019 TI - Peroxisome assembly factor-2, a putative ATPase cloned by functional complementation on a peroxisome-deficient mammalian cell mutant. AB - Rat peroxisome assembly factor-2 (PAF-2) cDNA was isolated by functional complementation of peroxisome deficiency of a mutant CHO cell line, ZP92, using transient transfection assay. This cDNA encodes a 978-amino acid protein with two putative ATP-binding sites. PAF-2 is a member of a putative ATPase family, including two yeast gene products essential for peroxisome assembly. A stable transformant of ZP92 with the cDNA was morphologically and biochemically restored for peroxisome biogenesis. Fibroblasts derived from patients deficient in peroxisome biogenesis (complementation group C) were also complemented with PAF-2 cDNA, indicating that PAF-2 is a strong candidate for the pathogenic gene of group C peroxisome deficiency. PMID- 7493020 TI - The VITESSE algorithm for rapid exact multilocus linkage analysis via genotype set-recoding and fuzzy inheritance. AB - As genetic marker maps have improved, multipoint linkage analysis has become a crucial part of all disease mapping studies. Paradoxically, multipoint lod scores become increasingly difficult to compute, particularly as the numbers of markers, marker alleles and untyped people increase. We have solved this problem by using a novel set-recording scheme to recode each person's genotype and 'fuzzy inheritance' to infer transmission probabilities. Our approach is implemented in a memory-efficient computer program, VITESSE, for extremely rapid computation of exact multipoint likelihoods. VITESSE enables fast and precise multipoint mapping of disease loci with highly polymorphic markers. PMID- 7493021 TI - Transforming growth factor-beta 3 is required for secondary palate fusion. AB - Mice lacking TGF-beta 3 exhibit an incompletely penetrant failure of the palatal shelves to fuse leading to cleft palate. The defect appears to result from impaired adhesion of the apposing medial edge epithelia of the palatal shelves and subsequent elimination of the mid-line epithelial seam. No craniofacial abnormalities were observed. This result demonstrates that TGF-beta 3 affects palatal shelf fusion by an intrinsic, primary mechanism rather than by effects secondary to craniofacial defects. PMID- 7493022 TI - Abnormal lung development and cleft palate in mice lacking TGF-beta 3 indicates defects of epithelial-mesenchymal interaction. AB - A broad spectrum of biological activities has been proposed for transforming growth factor-beta 3 (TGF-beta 3). To study TGF-beta 3 function in development, TGF-beta 3 null mutant mice were generated by gene-targeting. Within 20 hours of birth, homozygous TGF-beta 3-/- mice die with unique and consistent phenotypic features including delayed pulmonary development and defective palatogenesis. Unlike other null mutants with cleft palate, TGF-beta 3-/- mice lack other concomitant craniofacial abnormalities. This study demonstrates an essential function for TGF-beta 3 in the normal morphogenesis of palate and lung, and directly implicates this cytokine in mechanisms of epithelial-mesenchymal interaction. PMID- 7493023 TI - An integrated metric physical map of human chromosome 19. AB - A metric physical map of human chromosome 19 has been generated. The foundation of the map is sets of overlapping cosmids (contigs) generated by automated fingerprinting spanning over 95% of the euchromatin, about 50 megabases (Mb). Distances between selected cosmid clones were estimated using fluorescence in situ hybridization in sperm pronuclei, providing both order and distance between contigs. An average inter-marker separation of 230 kb has been obtained across the non-centromeric portion of the chromosome. Various types of larger insert clones were used to span gaps between contigs. Currently, the map consists of 51 'islands' containing multiple clone types, whose size, order and relative distance are known. Over 450 genes, genetic markers, sequence tagged sites (STSs), anonymous cDNAs, and other markers have been localized. In addition, EcoRI restriction maps have been generated for > 41 Mb (approximately 83%) of the chromosome. PMID- 7493025 TI - Mutations in the cardiac myosin binding protein-C gene on chromosome 11 cause familial hypertrophic cardiomyopathy. AB - Familial hypertrophic cardiomyopathy (FHC) is an autosomal dominant disorder manifesting as cardiac hypertrophy with myocyte disarray and an increased risk of sudden death. Mutations in five different loci cause FHC and 3 disease genes have been identified: beta cardiac myosin heavy chain, alpha tropomyosin and cardiac troponin T. Because these genes encode contractile proteins, other FHC loci are predicted also to encode sarcomere components. Two further FHC loci have been mapped to chromosomes 11p13-q13 (CMH4, ref. 6) and 7q3 (ref. 7). The gene encoding the cardiac isoform of myosin binding protein-C (cardiac MyBP-C) has recently been assigned to chromosome 11p11.2 and proposed as a candidate FHC gene. Cardiac MyBP-C is arrayed transversely in sarcomere A-bands and binds myosin heavy chain in thick filaments and titin in elastic filaments. Phosphorylation of MyBP-C appears to modulate contraction. We report that cardiac MyBP-C is genetically linked to CMH4 and demonstrate a splice donor mutation in one family with FHC and a duplication mutation in a second. Both mutations are predicted to disrupt the high affinity, C-terminal, myosin-binding domain of cardiac MyBP-C. These findings define cardiac MyBP-C mutations as the cause of FHC on chromosome 11p and reaffirm that FHC is a disease of the sarcomere. PMID- 7493024 TI - Germline mutations of the BRCA1 gene in breast and ovarian cancer families provide evidence for a genotype-phenotype correlation. AB - Mutations in the BRCA1 gene, discovered in 1994, are associated with an 80-90% lifetime risk of breast cancer. We have analysed 60 families with a history of breast and/or ovarian cancer for germline mutations in BRCA1. Twenty-two different mutations were detected in 32 families (53%), of which 14 are previously unreported. We observed a significant correlation between the location of the mutation in the gene and the ratio of breast to ovarian cancer incidence within each family. Our data suggest a transition in risk such that mutations in the 3' third of the gene are associated with a lower proportion of ovarian cancer. Haplotype analysis supports previous data which suggest some BRCA1 mutation carriers have common ancestors; however, we have found at least two examples where recurrent mutations appear to have arisen independently. PMID- 7493027 TI - Familial cylindromatosis (turban tumour syndrome) gene localised to chromosome 16q12-q13: evidence for its role as a tumour suppressor gene. AB - The human skin is a complex organ composed of the surface epidermis, the subjacent dermis (in which blood vessels, lymphatics and nerves are located) and the skin appendages. The latter include hair follicles, sebaceous glands (which secrete lipids that may serve as a permeability barrier, emollient or antimicrobial agent), apocrine glands (which secrete scents) and eccrine glands (which produce sweat for temperature control). Hereditary cylindromatosis (MIM 123850) is a rare autosomal dominant disease characterised by the development of multiple neoplasms originating from the skin appendages. These neoplasms have been termed cylindromas due to their characteristic microscopic architecture and are believed to exhibit apocrine or eccrine differentiation. We have carried out a genome search using two families with this disease, which has provided strong evidence for linkage of cylindromatosis to loci on chromosome 16q12-q13. Using markers close to the cylindromatosis gene, consistent loss of the wild-type allele was observed in 19 tumours from four individuals in the two families, indicating that the gene is likely to be a tumour suppressor gene. PMID- 7493026 TI - Cardiac myosin binding protein-C gene splice acceptor site mutation is associated with familial hypertrophic cardiomyopathy. AB - Familial hypertrophic cardiomyopathy (FHC) is an autosomal dominant disease characterized by a ventricular hypertrophy predominantly affecting the interventricular septum and associated with a large extent of myocardial and myofibrillar disarray. It is the most common cause of sudden death in the young. In the four disease loci found, three genes have been identified which code for beta-myosin heavy chain, cardiac troponin T and alpha-tropomyosin. Recently the human cardiac myosin binding protein-C (MyBP-C) gene was mapped to chromosome 11p11.2 (ref. 8), making this gene a good candidate for the fourth locus, CMH4 (ref. 5). Indeed, MyBP-C is a substantial component of the myofibrils that interacts with several proteins of the thick filament of the sarcomere. In two unrelated French families linked to CMH4, we found a mutation in a splice acceptor site of the MyBP-C gene, which causes the skipping of the associated exon and could produce truncated cardiac MyBP-Cs. Mutations in the cardiac MyBP-C gene likely cause chromosome 11-linked hypertrophic cardiomyopathy, further supporting the hypothesis that hypertrophic cardiomyopathy results from mutations in genes encoding contractile proteins. PMID- 7493028 TI - Mutation in the iron responsive element of the L ferritin mRNA in a family with dominant hyperferritinaemia and cataract. AB - The synthesis of ferritin, the iron-storing molecule, is regulated at the translational level by iron through interaction between a cytoplasmic protein, iron regulatory protein (IRP), and a conserved nucleotide motif present in the 5' non-coding region of all ferritin mRNAs--the iron responsive element (IRE). This region forms a stem-loop structure and when the supply of iron to the cells is limited, the IRP is bound to IRE and represses ferritin synthesis. Ferritin is composed of a 24-subunit protein shell surrounding an iron core. The two types of subunit, H and L, are encoded by two genes located on chromosomes 11q13 and 19q13.1, respectively. Both genes are ubiquitously expressed but transcriptional regulation mediates tissue-specific changes in the H/L mRNA ratio and isoferritin profiles. We now report the identification of a single point mutation in the IRE of the L-ferritin mRNA in members from a family affected with dominantly inherited hyperferritinaemia and cataract. This mutation consists of an A to G change in the highly conserved CAGUGU motif that constitutes the IRE loop and mediates the high-affinity interaction with the IRP. We show that this mutation abolishes the binding of IRP in vitro and leads to a high constitutive, poorly regulated L-ferritin synthesis in cultured lymphoblastoid cells established from affected patients. This is, to our knowledge, the first mutation affecting the IRP-IRE interaction and the iron-mediated regulation of ferritin synthesis. We suggest that excess production of ferritin in tissues is responsible for the hyperferritinaemia and that intracellular accumulation of ferritin leads to cataract. PMID- 7493029 TI - M6P/IGF2R gene is mutated in human hepatocellular carcinomas with loss of heterozygosity. AB - The mannose 6-phosphate/insulin-like growth factor-II receptor (M6P/IGF2R) functions in the intracellular trafficking of lysosomal enzymes, the activation of the potent growth inhibitor, transforming growth factor beta 2, and the degradation of IGF2 (ref. 1), a mitogen often overproduced in tumours. We have recently shown that 70% of human hepatocellular tumours have loss of heterozygosity (LOH) at the M6P/IGF2R locus which maps to chromosome 6q26-q27 (ref. 8). Using a coarse screen, we have now identified point mutations in the remaining allele of 25% of human hepatocellular carcinomas (HCCs) with LOH. These mutations give rise to truncated receptor protein and significant amino acid substitutions, and provide evidence that the M6P/IGF2R gene functions as a tumour suppressor in human liver carcinogenesis. PMID- 7493030 TI - A mutation in the mucosal keratin K4 is associated with oral white sponge nevus. AB - White sponge nevus (WSN) is a benign autosomal dominant disorder which affects non-cornifying stratified squamous epithelia (MIM 193900) (ref. 1). Phenotypically it presents as white 'spongy' plaques (oral leukokeratoses), most commonly in the mouth but also reported in the esophagus and anogenital mucosa. Histologically, the plaques show evidence of hyperproliferation, acanthosis and tonofilament aggregation. These types of pathogenic changes are characteristic of many of the epidermal keratin disorders. Keratins are expressed in pairs by epithelial cells in a tissue and cell specific manner. The major differentiation specific keratins of the buccal mucosa, nasal, esophageal and anogenital epithelia are K4 and K13 (ref. 7). The tissue distribution and nature of the lesions in patients affected by WSN suggested that mutations in K4 and/or K13 might be responsible for this disorder. We have now confirmed this hypothesis and report here a three base-pair (bp) deletion in the helix initiation peptide of K4 in affected members from two families with this condition. PMID- 7493031 TI - Keratin 13 point mutation underlies the hereditary mucosal epithelial disorder white sponge nevus. AB - Although pathogenic keratin mutations have been well characterized in inherited epidermal disorders, analogous defects in keratins expressed in non-epidermal epithelia have yet to be described. White sponge nevus (WSN) is a rare autosomal dominant disorder of non-cornifying squamous epithelial differentiation that presents clinically as bilateral white, soft, thick plaques of the oral mucosa. Less frequently the mucous membranes of the nose, esophagus, genitalia and rectum are involved. Histopathological features, including epithelial thickening, parakeratosis, extensive vacuolization of the suprabasal keratinocytes and compact aggregates of keratin intermediate filaments (KIF) in the upper spinous layers, resemble those found in epidermal disorders due to keratin defects. We analysed a multigenerational family with WSN and found cosegregation of the disease with the keratin gene cluster on chromosome 17. We identified a missense mutation in one allele of keratin 13 that leads to proline substitution for a conserved leucine. The mutation occurred within the conserved 1A region of the helical rod domain, which is critical for KIF stability and is the site of most pathogenic keratin mutations. This mutation enlarges the spectrum of keratins with disease-causing defects to include mucosally expressed keratin 13, and extends the known keratin diseases to disorders of non-cornifying stratified squamous epithelia. PMID- 7493033 TI - Opitz syndrome is genetically heterogeneous, with one locus on Xp22, and a second locus on 22q11.2. AB - Opitz syndrome (OS, McKusick 145410) is a well described genetic syndrome affecting multiple organ systems whose cardinal manifestations include widely spaced eyes and hypospadias (Fig. 1). It was first reported as two separate entities, BBB syndrome, and G syndrome. However, subsequent reports of families in which the BBB and G syndrome segregated within a single kindred suggested that they were a single clinical entity. Although the original pedigrees were consistent with X-linked and autosomal dominant inheritance, male-to-male transmission in subsequent reports suggested that OS was inherited as an autosomal dominant trait. Here we report that OS is a heterogeneous disorder, with an X-linked and an autosomal locus. Three families were linked to DXS987 in Xp22, with a lod score of 3.53 at zero recombination. Five families were linked to D22S345 from chromosome 22q11.2, with a lod score of 3.53 at zero recombination. This represents the first classic multiple congenital anomaly syndrome with an X-linked and an autosomal form. PMID- 7493032 TI - Fibrillin-2 (FBN2) mutations result in the Marfan-like disorder, congenital contractural arachnodactyly. AB - Congenital contractural arachnodactyly (CCA) is an autosomal dominant disorder that is phenotypically similar to Marfan syndrome (MFS) and characterized by arachnodactyly, dolichostenomelia, scoliosis, multiple congenital contractures and abnormalities of the external ears. In contrast to MFS, CCA does not affect the aorta or the eyes. Two closely related genes, FBN1 located on chromosome 15q15-21.3 and FBN2 located at 5q23-31, encode large fibrillin proteins found in extracellular matrix structures called microfibrils. The MFS is caused by mutations in FBN1, while CCA has been genetically linked to FBN2 (refs 2, 5, 6). We now describe a pair of FBN2 missense mutations in two CCA patients that cause substitution of distinct cysteine residues in separate epidermal growth-factor like (EGF) repeats. Our study provides final proof of the association between FBN2 mutations and CCA pathology, thus establishing the role of the fibrillin-2 in extracellular matrix physiology and pathology. PMID- 7493034 TI - Fibroblast growth factor receptor 3 (FGFR3) transmembrane mutation in Crouzon syndrome with acanthosis nigricans. AB - Crouzon syndrome, an autosomal dominant condition characterized by craniosynostosis, ocular proptosis and midface hypoplasia, is associated with mutations in fibroblast growth factor receptor 2 (FGFR2) (refs 1-3). For example, we have identified 10 different mutations in the FGFR2 extracellular immunoglobulin III (IgIII) domain in 50% (16/32) of our Crouzon syndrome patients. All mutations described so far for other craniosynostotic syndromes with associated limb anomalies--Jackson-Weiss, Pfeiffer, and Apert--also occur in the extracellular domain of FGFR2, as well as FGFR1 for Pfeiffer syndrome. In contrast, only FGFR3 mutations have been reported in dwarfing conditions- achondroplasia, thanatophoric dysplasia, and hypochondroplasia. For achondroplasia, greater than 99% of mutations occur in the FGFR3 transmembrane domain. We now report the unexpected observation of a FGFR3 transmembrane domain mutation, Ala391Glu, in three unrelated families with Crouzon syndrome and acanthosis nigricans, a specific skin disorder of hyperkeratosis and hyperpigmentation. The association of non-dwarfing and even non-skeletal conditions with FGFR3 mutations reveals the potential for a wide range of FGFR pleiotropic effects as well as locus heterogeneity in Crouzon syndrome. Our study underscores the biologic complexity of the FGFR gene family. PMID- 7493035 TI - Cloning of the sulphamidase gene and identification of mutations in Sanfilippo A syndrome. AB - Sanfilippo A syndrome is one of four recognised Sanfilippo sub-types (A, B, C and D) that result from deficiencies of different enzymes involved in the lysosomal degradation of heparan sulphate; patients suffer from severe neurological disorders. The Sanfilippo syndrome sub-types are also known as mucopolysaccharidosis (MPS) type III (MPS-IIIA, B, C and D), and are part of the large group of lysosomal storage disorders. Each of the MPS-III types is inherited as an autosomal recessive disorder with considerable variation in severity of clinical phenotype. The incidence of Sanfilippo syndrome has been estimated at 1:24,000 in The Netherlands with MPS IIIA (MIM #252900) the most common. MPS-IIIA is the predominant MPS-III in the United Kingdom, and has a similar high incidence to that found in The Netherlands (E. Wraith, personal communication). There is a particularly high incidence of a clinically severe form of MPS-IIIA in the Cayman Islands with a carrier frequency of 0.1 (ref. 4). Due to the mild somatic disease compared to other MPS disorders there is difficulty in diagnosing mild cases of MPS-III, hence Sanfilippo syndrome may be underdiagnosed, especially in patients with mild mental retardation. Here, we report the isolation, sequence and expression of cDNA clones encoding the enzyme sulphamidase (EC 3.10.1.1). In addition, we report the chromosomal localisation of the sulphamidase gene as being 17q25.3. An 11-bp deletion, present in sulphamidase cDNA from two unrelated Sanfilippo A patients, is described. PMID- 7493036 TI - Autosomal recessive retinitis pigmentosa caused by mutations in the alpha subunit of rod cGMP phosphodiesterase. AB - Retinitis pigmentosa (RP) constitutes a group of genetically heterogeneous progressive photoreceptor degenerations leading to blindness and affecting 50,000 100,000 people in the U.S. alone. Over 20 different RP loci have been mapped, of which six have been identified. Three of these encode members of the rod photoreceptor visual transduction cascade: rhodopsin, the rod cGMP-gated cation channel alpha subunit, and the beta subunit of cGMP-phosphodiesterase (PDEB). As null mutations in PDEB cause some cases of RP and since both alpha and beta subunits are required for full phosphodiesterase activity, we examined the gene encoding the alpha subunit of cGMP phosphodiesterase (PDEA) in 340 unrelated patients with RP. We found three point mutations in PDEA in affected members of two pedigrees with recessive RP. Each mutation alters an essential functional domain of the encoded protein and likely disrupts its catalytic function. PDEA is the seventh RP gene identified, highlighting the extensive genetic heterogeneity of the disorder and encouraging further investigation into the role of other members of the phototransduction cascade in RP. PMID- 7493037 TI - Diphenylhexatriene as a fluorescent probe for monitoring low density lipoprotein peroxidation. AB - The use of the fluorescent probe diphenylhexatriene (DPH) for monitoring low density lipoprotein (LDL) peroxidation has been investigated. The DPH incorporation into LDL results in a high fluorescence signal which decreases with time after addition of cupric ions. A strong correlation was found between the decay of the DPH fluorescence signal and the appearance of the thiobarbituric reactive substances (TBARS). HPLC and spectrofluorometric analyses demonstrated that DPH is destroyed during the time course of the copper-induced LDL peroxidation. The decrease in DPH fluorescent signal is prevented by addition of EDTA, vitamin E and drugs which protect LDL against peroxidation such as probucol or calcium antagonists. The high fluorescence of DPH allows the use of very small quantities of LDL (less than 5 micrograms/ml LDL protein). We thus suggest that DPH could be of use for continuous monitoring of LDL autooxidation, especially for the in vitro testing of the protective effect of antioxidant compounds. PMID- 7493039 TI - Dynamics of free radical formation from the reaction of peroxides with haemproteins as studied by stopped-flow chemiluminescence. AB - Metal ions such as iron and copper are implicated in the generation of free radicals in vivo, but the dynamics of free radical production in the decomposition of hydrogen peroxide and hydroperoxides by iron complexes have not been shown unequivocally due to their rapid rates. In the present study, we have devised a stopped-flow chemiluminescence system and succeeded in elucidating the time course of the free radical formation in the decomposition of hydrogen peroxide and organic hydroperoxides by haemproteins. The rate of free radical formation was very much dependent on the metal complexes and peroxides. Methemoglobin and cytochrome c decomposed hydrogen peroxide slower than microperoxidase but generated free radicals continuously. Methyl linoleate hydroperoxide was decomposed more rapidly than hydrogen peroxide. The chemiluminescence intensity was directly proportional to the amount of hydroperoxide, and the chemiluminescence progress curve reflected the time course of free radical flux. These results show that this stopped-flow chemiluminescence system is suitable to follow the radical flux from the decomposition of peroxides by metal ions. PMID- 7493038 TI - A new fluorescence method for the continuous determination of surface lipid oxidation in lipoproteins and plasma. AB - We report on a new method for the determination of lipid oxidation in lipoproteins and plasma. The biological lipid system is preloaded with a fluorescent analog of phosphatidylcholine containing diphenylhexatriene (DPH) propionic acid covalently linked to the sn-2 position. When externally added, the respective phospholipid label (DPHPC) localizes to the surface monolayer of a lipoprotein. Under oxidative conditions (e.g. in the presence of Cu2+ ions) the fluorophore undergoes decomposition, resulting in a continuous decrease of fluorescence intensity which reflects the oxidation of a chemically defined phospholipid molecule with well defined localization. When incorporated into LDL particles, the kinetics of the decrease in DPHPC fluorescence intensity upon exposure to Cu2+ us very similar to that of conjugated diene accumulation. Furthermore, our assay can be applied to follow the oxidation of lipids in diluted serum and may also be developed into a suitable test system for clinical studies of susceptibility of plasma lipids to oxidation. PMID- 7493040 TI - Does manganese protect cultured human skin fibroblasts against oxidative injury by UVA, dithranol and hydrogen peroxide? AB - Reactive oxygen species (ROS) are involved in the mechanism of photoaging and carcinogenesis. Skin is endowed with antioxidant enzymes including superoxide dismutases (SOD): cytosolic copper zinc SOD and mitochondrial manganese SOD. The aim of our study was to estimate the protective effect of manganese against oxidative injury on cultured human skin fibroblasts. Dithranol, hydrogen peroxide and UV-A radiation (375 nm) were employed as oxidative stressors. The supply of manganese chloride produced an increase in cellular content of this element up to 24 fold without concomitant elevation of MnSOD activity. Nevertheless, manganese protects cells against two of the three ROS generating systems assessed, namely hydrogen peroxyde and UV-A. This protective effect depends on the concentration of manganese in the medium, 0.1 mM and 0.2 mM protect against UVA cytotoxicity, only 0.2 mM protects against H2O2 cytotoxicity. PMID- 7493041 TI - Use of nitroxides as NMR contrast enhancing agents for joints. AB - NMR imaging is a well-established technology for obtaining cross-sectional anatomic pictures of organs and tissues. In addition, NMR can provide valuable information about the physiologic state of organs and tissues, especially, as a consequence of cellular injury. With this in mind, NMR in combination with gadolinium-based contrast enhancing agents has been used to assist in the detection of abnormalities to joints as well as to evaluate the status of damage resulting from an injury to this site. We describe the synthesis of a new nitroxide, which is bioresistant to the one-electron reduction mediated by superoxide in the presence of cysteine. This model mimics the reduction of nitroxides by extracellular secretion of superoxide by PMA-stimulated neutrophils. With this nitroxide, we found, in the range from 15 to 17.5 mumoles, enhancement of an NMR image in the knee joint of rabbits. Of interest is the finding that the contrast image remained for at least 90 minutes. These results demonstrate the utility of nitroxides as contrast enhancing agents for NMR imaging of joints. PMID- 7493042 TI - The free radical scavenger, alpha-lipoic acid, protects against cerebral ischemia reperfusion injury in gerbils. AB - alpha-Lipoic acid (thioctic acid) was tested for its neuroprotective activity in a Mongolian gerbil model of forebrain ischemia/reperfusion. Adult gerbils were treated for 7 days with two intraperitoneal injections per day of alpha-lipoic acid (20 mg/kg), vehicle or saline and on the 7th day the animals were subjected to 5 min of forebrain ischemia. Ischemic injury was assessed by monitoring the increases in locomotor activity and from the extent of damage to the CA1 hippocampal pyramidal cell layer after 5 days of recovery. By both criteria, alpha-lipoic acid was neuroprotective against ischemia/reperfusion evoked cerebral injury. PMID- 7493044 TI - Effects of superoxide on nitric oxide-dependent N-nitrosation reactions. AB - Recent studies have demonstrated that nitric oxide (NO) in the presence of superoxide (O2-) may mediate mutagenesis via the N-nitrosation of DNA bases followed by nitrosative deamination to yield their hydroxylated derivatives. We have found that phorbol myristate acetate (PMA)-activated extravasated rat neutrophils (PMNs) will N-nitrosate 2,3-diaminonaphthalene (DAN) to yield its highly fluorescent nitrosation product 2,3-naphthotriazole (triazole) via the L arginine dependent formation of NO. Addition of SOD enhanced triazole formation suggesting that O2- production may inhibit the N-nitrosating activity and thus the mutagenic activity of inflammatory PMNs. The objective of this study was to assess the role of superoxide as a modulator of NO-dependent N-nitrosation reactions using PMA-activated PMNs as well as a chemically defined-system that generates both NO and superoxide. We found that PMA-activation of PMNs reduced found that PMA-activation of PMNs reduced the amount of N-nitrosation of DAN by approximately 64% when compared to non-stimulated cells (450 vs. 1250 nM). Addition of SOD but not inactivated SOD or catalase to PMA-activated PMNs enhanced the formation of triazole by approximately 4-fold (1950 nM). In addition, we found that the NO-releasing spermine/NO adduct (Sp/NO; 50 microM) which produces approximately 1.0 nmol NO/min generated approximately 8000 nM of triazole whereas the combination of Sp/NO and a superoxide generator (hypoxanthine/xanthine oxidase) that produces approximately 1.0 nmol O2-/min reduced triazole formation by 90% (790 nM). Addition of SOD but not catalase restored the N-nitrosating activity. We conclude that equimolar fluxes of superoxide react rapidly with NO to generate products that have only limited ability to N-nitrosate aromatic amino compounds and thus may have limited ability to promote mutagenesis via the nitrosative deamination of DNA bases. PMID- 7493043 TI - Nitric oxide-dependent N-nitrosating activity of rat pleural mesothelial cells. AB - Recent studies have demonstrated that nitric oxide (NO)-derived N-nitrosating agents may promote mutagenesis and carcinogenesis from the nitrosative deamination of DNA bases via the formation of nitrosamine intermediates. The objective of this study was to determine if pleural mesothelial cells (PMC) stimulated with proinflammatory cytokines could promote the N-nitrosation of a primary aromatic amine via the L-arginine-dependent formation of NO-derived N nitrosation agents. N-nitrosating activity was determined by measuring the N nitrosation of a model amine, 2,3-diaminonapthalene, to yield its fluorescent triazole (1-naptho-2,3-triazole) derivative. Results show that specific combinations of TNF, IL-1, interferon gamma, and LPS significantly increased N nitrosating activity. There was a significant positive correlation between nitrite plus nitrate and triazole production. Triazole formation was inhibited by NG-nitro-L-arginine methyl ester, suggesting that triazole was derived from L arginine-dependent formation of NO. These data indicate that PMC have the capacity to promote the N-nitrosation of primary aromatic amines via the formation of NO. PMID- 7493045 TI - The chemical origin of free radicals in coffee and other beverages. AB - Sugars or carbohydrates are identified as the source of free radicals in coffees, ersatz coffees, a number of other food flavouring and colouring agents formed by processes involving heating, and in beers and stouts. The radicals are not derived from phenolic constituents, in contrast to those in wine, and are unlikely to be due solely to the occurrence of Maillard reactions. PMID- 7493046 TI - Cross-linked hemoglobins being well into clinical trials, increasing research efforts are now on a second generation red blood cell substitute based on encapsulated hemoglobin. PMID- 7493047 TI - Role of nitric oxide scavenging in peripheral vasoconstrictor response to beta beta cross-linked hemoglobin. AB - Transfusion with many crosslinked hemoglobin solutions causes an increase in arterial pressure that may be mediated by scavenging of nitric oxide (NO). If so, we postulated that inhibiting synthesis of NO after hemoglobin transfusion would fail to cause vasoconstriction ordinarily seen with such inhibition. In pentobarbital anesthetized cats, we tested whether administration of the NO synthase inhibitor, NG-nitro-L-arginine methyl ester (L-NAME), produced peripheral vasoconstriction after isovolemic exchange transfusion with hemoglobin to the same extent as occurs with L-NAME infusion in time controls and in controls matched for reduced hematocrit (17%) with albumin transfusion. Bovine hemoglobin was treated aerobically with bis-(3,5-dibromosalicyl)fumarate to produce beta beta-81 lysine crosslinks. Hemoglobin exchange transfusion increased mean arterial blood pressure and there was no further increase after L-NAME. In contrast, L-NAME increased pressure in the time controls and albumin controls. Hemoglobin transfusion decreased intestinal and renal blood flow, and there was no further decrease after L-NAME. In contrast, L-NAME decreased intestinal and renal blood flow in time controls and albumin controls. With L-NAME pretreatment in a separate group of cats, there was little further increase in arterial pressure or visceral vasoconstriction after hemoglobin transfusion. We conclude that the increase in arterial blood pressure after isovolemic crosslinked hemoglobin transfusion is best explained by scavenging of NO in intestinal and renal vascular beds. PMID- 7493048 TI - Nitrosyl hemoglobin formation in-vivo after intravenous administration of a hemoglobin-based oxygen carrier in endotoxemic rats. PMID- 7493049 TI - Review of modified hemoglobin research at Letterman: attempts to delineate the toxicity of cell-free tetrameric hemoglobin. AB - In the final two years, June 1991 to June 1993, of the Letterman Army Institute of Research, a variety of cell, tissue, organ, and animal systems were used to explore the toxicities of model hemoglobin (Hb) solutions produced in the sterile Hb production facility. Human mononuclear cells release TNF alpha and Il-8 when exposed to chromatographically purified human Hb (HbA0). Mixed cultures of fetal mouse neurons and glial cells exhibit neuronal death with exposure to HbA0 in a dose and time dependent manner while the glial cells are not injured. Isolated perfused rabbit hearts were used to explore the reversibility of coronary vasoconstriction after Hb and cyanomet-Hb administration, and deferoxamine was shown to partially protect that reversibility. In rabbits HbA0 and human Hb cross linked with bis(3,5-dibromosalicyl) fumarate (alpha alpha Hb) caused hypertension and pulmonary arteritis. In swine, HbA0 and alpha alpha Hb caused systemic and pulmonary hypertension and a doubling of the vascular resistance that was equivalent to that seen with inhibition of nitric oxide synthesis. Elevations of creatine kinase and lactic dehydrogenase activity were observed after Hbs were infused, but not after blockade of nitric oxide synthesis. Acute renal failure seen after administration HbA0, did not appear after alpha alpha Hb. Infusion of cyanomet-alpha alpha Hb did not cause the increased vascular resistance seen after alpha alpha Hb. The infusion of 1-arginine or nitroglycerine with alpha alpha Hb did not prevent the increased vascular resistance and decreased cardiac output or allow the increased oxygen carrying capacity provided by Hb in the plasma from translating into improved oxygen delivery or improved oxygen consumption. PMID- 7493051 TI - Hemoglobin mediated vasoactivity in isolated vascular rings. AB - To define hemoglobin-based oxygen carrier (HBOC) vasoactivity, a preliminary study with isolated vascular ring preparations was performed. Transverse ring segments of pulmonary artery and aorta were obtained from male SD rats, mounted in a tissue bath containing Krebs buffer (pH = 7.4, 37C) and instrumented to isometrically record changes in circumferential tension. Norepinephrine (NE) 0.1 70uM significantly increased vascular tension (N = 8, P < 0.01). Human stroma free Hb (SFH) at 1.8nM to 1.8uM Hb caused a dose dependent increase in isometric tension while methemoglobin 2nM-1.2uM had no significant effect (P > 0.05). Vessels preconstricted with SFH significantly relaxed (P < 0.05) with sodium nitroprusside, glyceryl trinitrate, and isoproteranol. SFH at micromolar concentration causes substantial vasoconstriction in isolated vessel preparations. The mechanism of SFH mediated vasoactivity appears to involve Hb interaction with endothelium derived nitric oxide and was reversible with nitrogenous vasodilators and a beta receptor agonist indicating pharmacologic modulation may be possible. PMID- 7493052 TI - Intramolecular cross-linking of oxy hemoglobin by bis sulfosuccinimidyl suberate and sebacate: generation of cross-linked hemoglobin with reduced oxygen affinity. AB - The sulfosuccinimidyl esters of suberic and sebacic acids readily introduce intramolecular crosslinks into oxy HbA at pH 7.4, the relative efficiency of crosslinking by the suberate ester being slightly higher than that of sebacate. Nearly quantitative intramolecular crosslinking of HbA (0.5 mM) is achieved at pH 7.4 and 4 degrees C by using 5 and 10 fold molar excess of the suberic and sebacic acid, respectively. In contrast to the facile crosslinking reaction seen with the bis sulfosuccinimidyl sebacate, bis sulfosuccinimidyl sebacate and bis (3:5 dibromo salicyl) sebacate did not introduce any crosslinking into HbA despite the fact that the 'crosslinking arm' of the two bifunctional reagents is the same. The discrepant reactivity of the two reagents demonstrates the 'steering' influence of the negative charge of the leaving group of the reagent, namely sulfo succinimidyl moiety to specific domains of HbA rich in positively charged groups. A second advantage is also anticipated in the use of the sulfosuccinimidyl esters of aliphatic dicarboxylic acids. We speculate that the intermediate in the crosslinking reaction mimics the structural aspects of the low oxygen affinity 'psuedo crosslinked Hb'. Conversion of the low oxygen affinity 'psuedocrosslinked Hb' into crosslinked Hb by the formation of the second isopeptide bond may lead to the 'freezing in' of the elements of low oxygen affinity structure. Consistent with this speculation, the suberate crosslinked Hb indeed exhibited low oxygen affinity even though the crosslinking reaction was carried out in the oxy state.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7493050 TI - The autoxidation of alpha alpha cross-linked hemoglobin: a possible role in the oxidative stress to endothelium. AB - The aim of the present study was to investigate the role of hemoglobin autoxidation in the induction of endothelial heme oxygenase (HO), an inducible "stress" protein which is responsible for heme catabolism. Porcine aortic endothelial cells were incubated for six hours in the presence of 60 microM unmodified hemoglobin (HbA0), hemoglobin cross-linked between the alpha chains with bis-(3,5-dibromosalicyl) fumarate (alpha alpha Hb) or cyanomet-alpha alpha hemoglobin (CNmet alpha alpha Hb). Microsomal HO content increased 4.1-fold in the presence of alpha alpha Hb, 2.7-fold with HbA0 and 1.8-fold with CNmet alpha alpha Hb over the control value. The rates of methemoglobin formation exhibited a linear relationship over the time of incubation (r = 0.94) and the apparent rate constant was 1.8-fold higher for alpha alpha Hb (0.023 h-1) than HbA0 (0.013 h 1). In addition, a linear relationship was obtained by plotting the rates of autoxidation of hemoglobins versus the HO activity (r = 0.99). When cells were incubated with 100% methemoglobin, HO activity increased 5.0-fold and 4.7-fold for HbA0 and alpha alpha Hb, respectively. Intracellular heme concentration, measured after 24 hours of incubation, was also significantly greater in the presence of alpha alpha Hb (52.6% over baseline) compared to HbA0 (10.8%) and CNmet alpha alpha Hb (15.3%) groups (p < 0.05). However, lactate dehydrogenase (LDH) release, measured as an index of endothelial cell injury, increased in all the hemoglobins examined: alpha alpha Hb, 33.8 +/- 1.1 U/l; HbA0, 38.5 +/- 3.5 U/l; CNmet alpha alpha Hb, 41.9 +/- 4.0 U/l; (control group, 19.4 +/- 2.8 U/l). We conclude that: 1) the higher rate of oxidation of alpha alpha Hb renders the molecule more susceptible to induce endothelial oxidative stress (HO induction); 2) the accelerated methemoglobin formation is directly correlated to intracellular HO content and endothelial heme uptake; 3) persistent cell injury suggests that other factors besides heme release may contribute to the hemoglobin mediated cytotoxicity. PMID- 7493053 TI - In vitro effect of dextran-benzene-tetra-carboxylate hemoglobin on human blood rheological properties. AB - While conducting pharmacological investigations into oxygen carriers, it is important to study the in vitro and in vivo rheological behavior of blood cells in the presence of such preparations. With regard to the original nature of human hemoglobin bound to benzene tetracarboxylate substituted dextran (Dex-BTC-Hb), it seemed necessary to study its rheological effect in a simulated in vitro hemorrhagic shock compensated by a blood substitute. The viscosity of substitutes was determined as well as several rheological parameters after 0, 3 and 6 hours incubation periods of red blood cells with substitutes: viscosity of blood substitute mixtures at different levels of plasma substitution erythrocyte aggregation of blood-substitute mixtures by determining the velocity of rouleau formation and the cohesion of rouleau network. This work yielded several observations: The viscosity of Dex-BTC-Hb was slightly higher than those of solutions of native Hb, Dex-BTC T10, Dextran 40 (Plasmacair, modified fluid gelatin (Plasmion and hydroxyethyl starch 200 (Elohes). The substitution of a blood volume with Dex-BTC-Hb, corresponding to a compensated 45% hemorrhagic shock, slightly increased the viscosity of hemodiluted blood as compared to other substitutes. In the presence of Dex-BTC-Hb, the aggregation of erythrocytes appears to be increased as compared to standard solutions. Yet, the effect was close to that of Plasmion or Elohes. PMID- 7493054 TI - A comparative study of the accurate measurement of endotoxin in liposome encapsulated hemoglobin. AB - We have examined three different methods of endotoxin determination utilizing the Limulus Amebocyte Lysate (LAL) assay to accurately determine endotoxin levels in Liposome Encapsulated Hemoglobin (LEH), 1) the gel-clot method, 2) chromogenic spectroscopic-based LAL, and 3) the turbidimetric method which determines endotoxin levels in solutions based on the time needed to reach a specific degree of turbidity. Both the chromogenic and turbidimetric methods require significant dilution of the LEH preparation before accurate measurement can be made. We have tested the levels of endotoxin in LEH solutions using these methods and measured LEH, liposome, and hemoglobin samples spiked with known amounts of endotoxin. A comparison of the three methods shows that the absolute value of endotoxin measured in LEH by the three methods can vary significantly. However, within any one assay the spiked amount of endotoxin in the sample can be accurately measured. The accuracy of these methods may also be complicated by the binding of endotoxin to LEH. This was evident by mixing free endotoxin with LEH followed by centrifugation to separate the LEH. Biological activity of endotoxin bound to LEH was measured by exposure to RAW264.7 followed by the expression of tumor necrosis factor. PMID- 7493055 TI - Effect and safety of liposome-encapsulated hemoglobin neo red cells (NRCs) as a perfusate for total cardiopulmonary bypass. AB - We developed a liposome encapsulated hemoglobin named Neo Red Cells (NRC), NRC (1) readily circulates, (2) shows a high oxygen transport efficiency, and (3) has a strong capsule membrane. In this study, we evaluated the effect of NRC as a priming solution for total cardiopulmonary bypass (TCPB) using a dog. The TCPB was started after removal of 57.1-73.3% of autologous blood and continued for 7 hours. During TCPB using NRC, the vascular resistance (VR) decreased to 1/4 of VR when red blood cells (RBC) were used. This change suggests that NRC, the viscosity of which is lower than that of RBC, reduced the load on the circulation system. The oxygen volume delivered by NRC was higher than that delivered by RBC, resulting in a greater oxygen consumption with NRC. During TCPB using NRC, the serum LDH level was lower than that using RBC. So we concluded that NRC compensated for the reduction in the oxygen transport ability, which is a disadvantage of dilution TCPB, and further increased the circulation improving effect and anti-hemolytic effect, which are advantages of the procedure. It, thus, enhanced both the safety and effectiveness of dilution TCPB. PMID- 7493056 TI - Characterization of neo red cells (NRCs), their function and safety in vivo tests. AB - The Neo Red Cells (NRCs), a new type of artificial oxygen carriers have been developed and investigated for oxygen transport capacity, hemodynamics and safety in experimental animals. Stroma free hemoglobin (Hb) prepared from outdated human red blood cells was encapsulated together with inositol hexaphosphate (as an allosteric effector), coenzyme and substrates for reducing metHb back to Hb in liposomes under an aseptic condition. The NRCs were subsequently coated with polyethylene glycol bound to phosphatidylethanolamine as surface modifier to prevent aggregation of NRCs in plasma. The metHb formation was reduced from 1%/hr to 0.37%/hr by the addition of metHb reduction system. The blood pressure increased transiently during injection, and then immediately returned to pre injection levels. The efficiency of the NRCs in tissue oxygenation and recovery from anemia was studied in rabbits which had been made severely anemic by drawing 85% of their blood and replacing it with NRCs. All of the animals infused with NRCs recovered to pre-anemic conditions within 6-8 hr and survived until they were sacrificed, 6 months after the exchange transfusion. Our observations suggest that the NRCs are pathogen free, efficient oxygen carriers without causing serious adverse reactions, with controllable metHb formation in vivo and during storage. PMID- 7493057 TI - Complement activation by liposome-encapsulated hemoglobin in vitro: the role of endotoxin contamination. AB - Incubation of liposome-encapsulated Hb (LEH) with rat serum at 37 degrees C led to accelerated decay of serum hemolytic complement (C) activity (CH50/ml). Empty liposomes (L) caused less decrease of CH50/ml, whereas free Hb had no effect on C activity. The LEH- and L-induced increases in C consumption were unlikely a consequence of endotoxin (LPS) contamination, as spiking of rat serum with LPS caused reduction in C only at levels significantly higher than those detectable in LEH or L. LPS-induced C consumption was not potentiated by free hemoglobin. PMID- 7493058 TI - Perfluorocarbons in the twenty-first century: clinical applications as transfusion alternatives. AB - The risks of allogeneic transfusion are well known to physicians and have prompted a search for alternatives. Perfluorocarbons were introduced into clinical trials in the early 1980's with the hope that these products would develop into acceptable blood substitutes. Unfortunately, the limited potency, short half-life, and potential toxicity of these early formulations coupled with unrealistic expectations for efficacy prevented the perfluorocarbons from playing a significant role in transfusion medicine. Recent changes in formulation to improve efficacy and eliminate toxicity have stimulated renewed interest in perfluorocarbons as alternatives to allogeneic transfusion. Our recent work has focused on the role of perfluoroctylbromide (PFOB), a second generation perfluorocarbon, as an adjunct to autologous transfusion and acute normovolemic hemodilution (ANH), rather than as a total blood substitute. Initial animal experiments have shown the ability of small doses of PFOB to maintain oxygen delivery without the need for blood transfusion in the setting of hemodilution. Our presentation will focus on subsequent clinical work using PFOB as an alternative to both allogeneic and autologous blood transfusion during ANH. We believe that perfluorocarbons such as PFOB will have a significant role in the future as one of several, additive alternatives to blood transfusion. PMID- 7493059 TI - Use of Oxygent, a perfluorochemical-based oxygen carrier, as an alternative to intraoperative blood transfusion. AB - Oxygent is a stable concentrated perfluorochemical (PFC) emulsion being developed for use as a temporary oxygen carrier. In this application, PFC emulsions can be used to augment oxygen delivery during acute blood loss and thereby provide a margin of safety during hemodilution and surgical anemia. PFCs simply dissolve oxygen in direct proportion to its partial pressure. The oxygen transported by a PFC emulsion is present in the plasma compartment and is therefore easily extracted and consumed by the tissues. Preclinical and clinical studies have demonstrated that a relatively low dose (1.35 g PFC/kg) of Oxygent can support oxygen delivery despite ongoing blood loss. Clinical safety studies in 57 healthy, conscious volunteers and in 30 anesthetized surgical patients have been completed. In these studies, there were no hemodynamic changes or vasoconstriction and cardiac output increased normally in response to hemodilution. Two transient side effects were observed, but only in the high dose (1.8 g PFC/kg) group: a 1-1.5 degrees C increase in body temperature (at 4-6 hours), and a moderate decrease in platelet count (mean nadir approximately 130,000/microL by 2-3 days) without any bleeding complications. Oxygent is presently being evaluated as an alternative to allogeneic blood transfusion in patients undergoing medium- to high-blood-loss surgical procedures. PMID- 7493060 TI - An overview on oxygen carriers in cancer therapy. PMID- 7493061 TI - Several methods utilized for the assessment of biocompatibility of perfluorochemicals. AB - An overview is presented on methods used to judge the effect of different perfluorochemicals (PFCs) on functional organ changes. By some of these methods the function of the reticuloendothelial system (RES) after a load with PFC was tested, either globally in form of the clearance of colloidal carbon particles or by a magnetometric method to test the phagocytosing function of macrophages within the liver of rats. An endotoxin test on survival of mice after the combined treatment with lipopolysaccharides of E. coli and PFCs showed the importance of a cumulative effect of multiple factors influencing phagocytosis. Other methods examined the influence of PFCs on lipid peroxidation during reperfusion of the ischemic intestine or on the effects of hyperbaric oxygen on the lung. The half-life (mean dwell time) of PFCs within organs with a high storage rate could optimally be determined by means of 19F magnetic resonance imaging in longitudinal studies within the same animal. An influence of PFCs on hepatocytes was tested by determining the time required for detoxifying pentobarbital as measured by the sleeping time (until reappearance of the righting reflex). By all these tests a strong dose-dependence was found, so that effects observed with a high load may become negligible with smaller dosages. PMID- 7493062 TI - Enhanced protoplast growth at the interface between oxygenated fluorocarbon liquid and aqueous culture medium supplemented with pluronic F-68. AB - Protoplasts isolated from cell suspensions of albino Petunia hybrida were grown for 10 d at the interface between aqueous culture medium and oxygenated perfluorodecalin. Protoplasts synthesised new cell walls and divided normally at the fluorocarbon/culture medium interface, the mean division frequency of protoplasts being increased by 37% (P < 0.05) in this system when the perfluorodecalin was saturated with oxygen prior to use. The mean division frequency of protoplasts was further increased to a maximum of 52% above control (P < 0.01) when the medium overlaying the oxygenated perfluorodecalin was supplemented with 0.01% (w/v) of the co-polymer surfactant, Pluronic F-68. These results demonstrate a beneficial and synergistic effect of supplementing protoplast culture systems with oxygenated perfluorodecalin and Pluronic F-68. PMID- 7493063 TI - Restoration of tumor oxygenation after cytotoxic therapy by a perflubron emulsion/carbogen breathing. AB - Female, Fisher 344 rats bearing 13762 mammary carcinoma implanted subcutaneously in a hind limb were treated with standard therapeutic single doses of antitumor treatments of several types including: 1) antitumor alkylating agents (cisplatin, cyclophosphamide); 2) natural products (adriamycin, taxol and etoposide); 3) antimetabolites (5-flourouracil); 4) hypoxic cell selective agents (mitomycin C, SR-4233) as well as 5) fractionated radiation therapy (3 Gray daily for 5 days). The oxygen levels in the tumors were measured in the absence of treatment and 24 hrs. after treatment using an Eppendorf p02 histograph. Fifty- to sixty-points were measured per tumor and 8-10 tumors comprised each group. The tumors were more hypoxic post treatment with every anticancer drug or radiation. The percent of p02 readings < or = 5 mmHg in the untreated tumors ranged from 85% (x-rays) to 59% (etoposide). Administration of the perflubron emulsion (8 ml/kg) and carbogen breathing (95% O2/5% CO2) increased the oxygenation of the tumors such that the percent of pO2 readings < or = 5 mmHg was 32% in the untreated controls and ranged from 27% (x-rays) to 56% (adria) in the treated tumors. These results indicate that administration of a perflubron emulsion/carbogen can increase the oxygen content of tumors when hypoxia is the result of cytotoxic therapy. PMID- 7493065 TI - The initial response of blood leukocytes to incubation with perfluorocarbon blood substitute emulsions. AB - Due to their enhanced oxygen carrying characteristics, perfluorocarbon emulsions are valuable adjuncts to coronary revascularization procedures. However, the effects of perfluorocarbon emulsions on white blood cell function are unclear. The purpose of this study was to determine the effects of three perfluorocarbon emulsions on the adhesion characteristics of leukocytes. Heparinized blood from donor rats was mixed with either Phosphate Buffered Saline (PBS), PFB-1, PFB-2 (both non-complement activating perflubron emulsions, Alliance Pharmaceutical Corp.) or Fluosol (20% w/v PFC, Alpha Therapeutic) in a ratio of one part emulsion to six parts blood. The blood-emulsion mixtures were incubated for ten minutes at 37 degrees C, then passed down nylon wool leukocyte adhesion columns. Blood samples were counted for: Leukocytes, Granulocytes and Lymphocytes. % Cell Adherence was calculated as: (1-[WBC-out]/[WBC-in]). We found that, compared to PBS control, the granulocyte adherence to the nylon fiber was significantly increased for both Fluosol-20 and PFB-1 (P < 0.05). In contrast, PFB-2 (designed for reduced cell surface activity) caused no significant change in leukocyte adhesion. In fact, a modest decrease in granulocyte adhesion was observed with PFB-2. These findings suggest a direct effect of some, but not all, PFC emulsions on blood leukocyte function. PMID- 7493064 TI - Increased tumor oxygenation and radiation sensitivity in two rat tumors by a hemoglobin-based, oxygen-carrying preparation. AB - The rat 13762 mammary carcinoma and the rat 9L gliosarcoma were grown subcutaneously in a hind limb of female, Fisher 344 rats. The oxygen content of the tumors was determined using an Eppendorf pO2 histograph. Fifty-to-sixty oxygen measurements were made per tumor and there were 8-to-10 animals per group. The percent of pO2 readings < or = 5 mmHg in the mammary carcinoma was 49%, this was decreased to 34% by administration of the hemoglobin preparation (8 ml/kg) and further decreased to 29% when carbogen (95% O2/5% CO2) breathing was added to administration of the hemoglobin preparation. The percent of pO2 readings < or = 5 mmHg in the gliosarcoma was 49%, this was decreased to 24% by administration of the hemoglobin preparation and further decreased to 0% when carbogen breathing was added to administration of the hemoglobin preparation. Therapeutic response was assessed over a single-dose range of radiation therapy (10, 20 and 30 Gray). The dose modifying factor produced by the hemoglobin preparation/air was 1.6 and by the hemoglobin preparation/carbogen was 2.7 in the rat 13762 mammary carcinoma. The dose modifying factor produced by the hemoglobin preparation/air was 1.9 and by the hemoglobin preparation/carbogen was 2.9 in the rat 9L gliosarcoma. Administration of a hemoglobin-based oxygen carrier reduced tumor hypoxia and increased tumor response to radiation therapy. PMID- 7493066 TI - Vasoreactivity of Fluosol perfluorocarbon emulsion vs. Earl's balanced salt/albumin perfusates in the isolated perfused rat lung. AB - We compare the vasoreactive responses of Fluosol perfluorocarbon emulsion versus Earl's balanced salt+albumin solution as perfusates in an isolated perfused rat lung. Comparisons were made with three separate vasoconstrictive stimuli: norepinephrine, and prostaglandin F2 alpha dose-response analysis, and generation of hypoxic pulmonary vasoconstriction. The hypoxia experiments were also performed with N-nitro-arginine added to the perfusates to block production of endogenous nitric oxide prior to the application of hypoxic ventilation challenge. The vasoconstrictive effects of norepinephrine, PGF2 alpha and hypoxia were significantly depressed in the perfluorocarbon perfused versus salt/albumin solution perfused lungs. With the addition of NNA to the perfusates, hypoxic pulmonary vasoconstriction is enhanced in both Earl's/albumin perfused as well as in the fluorocarbon perfused lungs. We can conclude that the pulmonary vasoconstrictive effects of norepinephrine, PGF2 alpha, and hypoxia are depressed in perfluorocarbon emulsion perfused, compared to salt-albumin solution perfused lungs. It is possible that perfluorocarbon emulsions interfere with normal endothelial function. As such, these compounds may be useful in examination of normal and abnormal endothelial function in various research applications. PMID- 7493067 TI - Automated high-performance liquid chromatographic assay for lesopitron, a novel anxiolytic, in human plasma using on-line solid-phase extraction. AB - A totally automated liquid chromatographic assay method based on a Prospekt solid phase extraction unit was developed for the analysis of lesopitron and its metabolite 5-hydroxylesopitron in human plasma. On-line solid-phase extraction of lesopitron, 5-hydroxylesopitron and its internal standard in human plasma was carried out using C2 cartridges. After washing, the test substances were eluted with mobile phase onto an ODS-2 Inertsil column and measured by fluorescence detection. The total time for one analysis was 25 min. The method developed was selective and linear in the concentration range from 1 to 40 ng/ml for both parent drug and metabolite. Recovery of lesopitron and 5-hydroxylesopitron were higher than 80% and the quantification limits were 1 ng/ml for both compounds. Coefficients of variation obtained for precision parameters were all below 14.5% and 13.9% for parent drug and metabolite, respectively. Good values of accuracy were also obtained. PMID- 7493068 TI - Column-switching high-performance liquid chromatographic system with a laser induced fluorimetric detector for direct, automated assay of salivary cortisol. AB - In order to measure human stress, an easy and rapid, fully automated method for the determination of cortisol in saliva has been developed, using column switching high-performance liquid chromatography with laser-induced fluorescence detection, which involves post-column labeling with sulfuric acid. The developed system requires only 0.1 ml of saliva, and a simple pretreatment consisting of dilution and filtration is sufficient. The column-switching system consisted of a Polymer-Coated Mixed-Functional silica (PCMF) column for deproteinization, and a CN column for frontal concentration and separation. An ODS column in place of the CN provided a better separation, but required a post-column make-up of water for safe reaction. Detection limit of cortisol was 8 fmol (signal-to-noise ratio = 3), which is adequate for routine determination of normal levels of cortisol (1 20 pmol/ml). The analysis time was about 40 min and reproducibility was excellent with an R.S.D. of less than 5%. PMID- 7493069 TI - Isolation, identification and determination of sulfadiazine and its hydroxy metabolites and conjugates from man and rhesus monkey by high-performance liquid chromatography. AB - The following metabolites of sulfadiazine (S) were isolated from monkey urine by preparative HPLC: 5-hydroxysulfadiazine (5OH), 4-hydroxysulfadiazine (4OH) and the glucuronide (5OHgluc) and sulfate conjugate of 5OH (5OHsulf). The compounds were identified by NMR, mass and infrared spectrometry and hydrolysis by beta glucuronidase. The analysis of S, the hydroxymetabolites (4OH, 5OH) and conjugates N4-acetylsulfadiazine (N4), 5OHgluc and 5OHsulf in human and monkey plasma and urine samples was performed using reversed-phase gradient HPLC with UV detection. In plasma, S and N4 could be detected in high concentrations, whereas the other metabolites were present in only minute concentrations. In urine, S, the metabolites and conjugates were present. The limit of quantification of the compounds in plasma varies between 0.2 and 0.6 microgram/ml (S 0.31, N4 0.40, 4OH 0.20, 5OH 0.37, 5OHgluc 0.33 and 5OHsulf 0.57 microgram/ml). In urine it varies between 0.6 and 1.1 micrograms/ml (S 0.75, N4 0.80, 4OH 0.60, 5OH 0.80, 5OHgluc 0.80 and 5OHsulf 1.1 micrograms/ml). The method was applied to studies with healthy human subjects and Rhesus monkeys. The metabolites 5OH, 5OHgluc and 5OHsulf were present in Rhesus monkey and not in man. Preliminary results of studies of metabolism and pharmacokinetics in Rhesus monkey and man are presented. PMID- 7493070 TI - Development of a high-performance liquid chromatographic method for the analysis of staurosporine. AB - Staurosporine (Stsp), a protein kinase inhibitor, has been found to have a differential effect on the proliferation of normal and transformed cells in vitro. Hence, Stsp might be used in cancer therapy to arrest normal proliferating cells in G1, while permitting tumor cells to continue proliferation. The patient could then be treated with a therapeutic agent of maximum toxicity for actively proliferating tumor cells. To facilitate investigations of Stsp in vivo, we have developed an HPLC method for measuring the levels of Stsp in blood. Using a rat model, plasma containing Stsp is treated with acetone to precipitate proteins and extract the Stsp. The acetone extract is then subjected to reversed-phase HPLC on a mu Bondapak C18 column. Using a linear elution gradient of acetonitrile containing trifluoroacetic acid, Stsp elutes as a sharp peak at ca. 35 min which can be detected by UV absorption at 292 nm. No blood or reagent components interfere with its quantification. The calibration curve, ranging from 0.1 to 2.0 micrograms Stsp, demonstrated a linear response to Stsp concentration having a correlation coefficient (r2) of 0.990. Precision analysis demonstrated that the method will yield results that are +/- 11.6% from the mean 95% (two standard deviations) of the time. This method was used to measure Stsp levels in plasma after administering an injection of 0.2 mg Stsp into the jugular vein of rats. No Stsp could be detected in the plasma 5 min after injection, even though enough Stsp was administered to be easily detectable if it was simply contained in the plasma. Thus, it is concluded that some compartment other than the plasma must adsorb the Stsp from the plasma and sequester it in vivo. PMID- 7493071 TI - Analysis of cisapride in neonatal plasma using high-performance liquid chromatography with a base-stable column and fluorescence detection. AB - A simple, selective, sensitive, and precise high-performance liquid chromatographic plasma assay for the prokinetic drug cisapride is described. Alkalinised samples of plasma (100 microliters) were extracted with 1.0 ml of 10% (v/v) isopropanol in chloroform, dried, redissolved in mobile phase and injected. Chromatography was performed at 20 degrees C by pumping a mobile phase of acetonitrile (370 ml) in pH 5.2, 0.02 M phosphate buffer (630 ml) at 1.0 ml/min through a C8 Symmetry column. Cisapride and the internal standard were detected by fluorescence monitoring at 295 nm (excitation) and 350 nm (emission), and were eluted 5 min and 8 min, respectively, after injection. Calibration plots in bovine serum albumin (3% w/v) were linear (r > 0.999) from 5 to 250 ng/ml. Intra day and inter-day precision (C.V.) was 9.5%, or less, and the accuracy was within 5.5% of the nominal concentration over the range 8-200 ng/ml. Total assay recovery was above 82%. Endogenous plasma components, the major cisapride metabolite (norcisapride), and other drugs used in neonatal pharmacotherapeutics did not interfere. PMID- 7493072 TI - A column-switching high-performance liquid chromatographic assay for a novel cytotoxic thioxanthone derivative (WIN 33377) in mouse plasma with toxicokinetic results from a mouse LD10 study. AB - WIN 33377 (I) is a member of a novel class of cytotoxic antitumor agents, 4 aminomethyl thioxanthone derivatives. A simple, rapid and reproducible method has been developed for the assay of I in mouse plasma using a high-performance liquid chromatographic method utilizing a column-switching technique. The method involves direct injection of buffered plasma to the extraction column for sample clean-up followed by switching onto an analytical column for analysis with UV detection at 256 nm. The method has demonstrated accuracy and precision over the range 10-2500 ng/ml using a 100-microliters plasma sample with a minimum quantifiable level at 10 ng/ml. Stability of mouse plasma samples was demonstrated after storage for 4 weeks at -15 to -20 degrees C, as was the ability of samples to be accurately quantified after a maximum of three freeze thaw cycles. Recovery was greater than 87% for the compound and the internal standard. The assay was accurate and reproducible with measured values lying within the limits of defined acceptance criteria. The utility of the method was demonstrated by analyzing plasma samples obtained from mice dosed with I as part of a pre-clinical safety study intended to assist in the design of a pharmacokinetically guided dose escalation strategy. PMID- 7493073 TI - High-performance liquid chromatographic determination of prolylcarboxypeptidase activity in monkey kidney. AB - A simple HPLC procedure for the determination of prolylcarboxypeptidase activity in monkey kidney was established with Cbz-Pro-Ala used as substrate. Decrease of the substrate and increase of the product were stoichiometrically related to each other. Heat treatment at 60 degrees C freed the enzyme preparation of contaminating activities. Data on substrate specificity and influence of inhibitors suggested this method was sensitive for the determination of prolylcarboxypeptidase without the use of a radioactive substrate. PMID- 7493074 TI - Heparin-immobilized polyhydroxyethylmethacrylate microbeads for cholesterol removal: a preliminary report. AB - Heparin-attached polyhydroxyethylmethacrylate (PHEMA) microbeads were investigated for specific removal of cholesterol from human and rabbit plasma. PHEMA microbeads were prepared by a suspension polymerization technique and activated by cyanogen bromide (CNBr) in an alkaline medium (pH 11.5). Heparin was then immobilized by covalent binding onto these microbeads. Cholesterol adsorption onto PHEMA microbeads containing two different amounts of immobilized heparin, i.e., 57.3 and 122.7 mg/g, from both hypercholesterolaemic human and rabbit plasma was investigated. The non-specific cholesterol adsorptions on the plain PHEMA microbeads were 0.47 mg/g and 0.30 mg/g from human and rabbit plasmas, respectively. About 35% and 32% of the cholesterol was removed from human and rabbit plasmas, respectively, when the heparin-immobilized PHEMA microbeads were used. PMID- 7493075 TI - Simultaneous determination of amphetamine, methamphetamine, 3,4 methylenedioxyamphetamine and 3,4-methylenedioxymethamphetamine in human hair by gas chromatography-mass spectrometry. AB - A procedure is presented for the simultaneous identification and quantification of amphetamine (AP), methamphetamine (MA), methylenedioxyamphetamine (MDA) and methylenedioxymethamphetamine (MDMA) in human hair. The method involves decontamination of hair with dichloromethane and warm water, heat-alkaline hydrolysis in the presence of deuterated internal standards, liquid-liquid extraction and gas chromatography-mass spectrometry after derivatization with pentafluoropropionic anhydride-pentafluoropropanol. The limit of detection for AP, MA and MDA was 0.05 ng/mg using a 50-mg hair sample; for MDMA it was 0.1 ng/mg. Coefficients of variation ranged from 7 to 18%. This assay has been successfully utilized in the evaluation of the deposition of the drugs in hair obtained from various parts of the anatomy of a stimulant abuser. PMID- 7493076 TI - Determination of muramic acid in organic dust by gas chromatography-mass spectrometry. AB - A method is described for the quantitation of muramic acid, a marker of bacterial peptidoglycan, in organic dust. House dust samples were hydrolysed in hydrochloric acid and then extracted with hexane to remove hydrophobic compounds. The aqueous phase was evaporated, heated in a silylation reagent to form trimethylsilyl derivatives, and analysed by gas chromatography--mass spectrometry. The muramic acid derivative gave two peaks upon injection into the gas chromatograph--mass spectrometer. Injection of 10 pg of the derivative gave a signal-to-noise ratio of 17 for the dominating peak when using selected ion monitoring in the electron impact mode, and a linear calibration curve was achieved upon analysis of samples containing 5-1500 ng of muramic acid. In a house dust sample, 40 ng of muramic acid was found per mg of dust; the coefficient of variation was 8.2% (n = 6, 1.2 mg of dust analysed). The described method is rapid and simple to apply, and should therefore become widely used for measuring peptidoglycan in many types of environmental samples, including organic dust. PMID- 7493077 TI - Sensitive high-performance liquid chromatographic assay method for the determination of guggulsterone in serum. AB - Guggulsterone (I) is a new hypolipidemic agent, being developed at CDRI (Lucknow, India). A sensitive high-performance liquid chromatographic assay in serum has been developed and validated for the determination of guggulsterone in serum for pharmacokinetic studies. This assay method consists of extraction of the drug with hexane from spiked human serum samples. Separation was achieved using C18 reversed-phase column coupled with photodiode array detector, and an acetonitrile water mixture as mobile phase. The method described herein is simple and has limit of quantitation of 10 ng/ml as compared to 200 ng/ml by the previous reported method. The standard curve was linear over the range of 10-1000 ng/ml in mobile phase as well as in normal human serum. Analytical recovery of I added to serum was > 90%. The reproducibility was determined by the inter- and intra-assay variations which were < 10%. PMID- 7493078 TI - High-performance liquid chromatography using electrochemical detection for the determination of prazosin in biological samples. AB - For the quantitation of prazosin a sensitive high-performance liquid chromatographic (HPLC) method was developed. This HPLC analysis method uses an electrochemical detection technique for the identification and quantitation of prazosin. In this assay the serum samples were deproteinized by using a simple acetonitrile precipitation technique that was followed by n-hexane extraction. Prazosin in the deproteinized serum sample was separated by an isocratic elution with an ODS Hypersil HPLC column (150 x 4.6 mm) using a mobile phase consisting of 0.05 M Na2HPO4-acetonitrile (60:40), pH 8.4. Prazosin that was eluted from the column was detected using a Coulochem II electrochemical detector. The precision of this assay method was assessed by performing inter- and intra-assay analyses by spiking prazosin free fetal bovine serum samples with 20 and 40 ng/ml concentrations of prazosin. In the intra-assay the recovery was 95.40 +/- 4.82% and 97.80 +/- 3.40%, respectively, for 20 and 40 ng/ml concentrations of prazosin that were used to spike the serum samples. This electrochemical detection HPLC assay method could be very useful in monitoring plasma levels of prazosin. PMID- 7493079 TI - Determination of sulphonamides in human urine by azo dye precolumn derivatization and micellar liquid chromatography. AB - A high-performance liquid chromatographic method for the determination of sulphonamides in urine is reported. The drugs (sulphadiazine, sulphaguanidine, sulphamethizole, sulphamethoxazole, and sulphathiazole) were diazotized with nitrite and coupled with N-(1-naphthyl)ethylenediamine dihydrochloride in a sodium dodecyl sulphate (SDS) micellar medium. Separation of the sulphonamide azo dyes was performed on a C18 column with a 0.05 M SDS-2.4% pentanol mobile phase, which permitted the direct injection of the urine samples. The limits of detection were in the 0.1-0.3 micrograms/ml range. PMID- 7493080 TI - Improved detection limit for catecholamines using liquid chromatography electrochemistry with a carbon interdigitated array microelectrode. AB - The detection limit of catecholamines can be lowered by using a carbon-based interdigitated array (IDA) microelectrode as a detector for liquid chromatography (LC). The IDA electrode is more sensitive than conventional glassy carbon electrodes due to the high current density caused by radial diffusion at each microband, and redox cycling between two microband arrays. Since the number of redox cycles increases at lower flow-rates, the carbon IDA is particularly useful for microbore LC. In an LC system with a 1-mm microbore column and a carbon IDA electrode, the peak height of dopamine (DA) and DOPAC did not decrease with decreasing flow-rate because of this redox cycling. A low detection limit of 5 fg (32 amol) and 9.6 fg (57 amol) was obtained for DA and DOPAC due to the high current density and low background noise level (0.1 pA) at the carbon IDA electrode. The total charge generated by oxidizing DA at the anodic array was more than the value calculated by assuming that all the DA molecules were oxidized. PMID- 7493081 TI - Determination of intracellular glutathione in human skeletal muscle by reversed phase high-performance liquid chromatography. AB - A chromatographic method for the specific determination of cellular low molecular mass thiols has been applied to human muscle tissue. The method is based on the derivatisation of thiols using monobromobimane, which is a specific reagent for the sulphydryl group. The glutathione and cysteine bimane adducts were separated by reversed-phase HPLC, whilst quantitation of the cysteine and glutathione adducts was achieved by fluorescence spectroscopy. The method was found to yield a quantitative recovery of glutathione (ca. 96%), to be sensitive (down to 20 pmol glutathione/per injection) and reveal a low intra-individual coefficient of variation (C.V. < 5%) of the glutathione concentrations in human skeletal muscle. The concentrations of reduced and total glutathione were 1320 +/- 37 mumol/kg wet weight (mean +/- S.E.M.) and 1525 +/- 66 mumol/kg wet weight, respectively. The method was also applied to tissues from nine healthy volunteers to determine if fluctuations in glutathione level occurred over a 24-h period. No diurnal variation of glutathione level in human skeletal muscle was observed. PMID- 7493082 TI - High-performance liquid chromatographic identification of eight constitutional disaccharides from heparan sulfate isomers digested with heparitinases. AB - Identification with specific heparan sulfate-lyases, heparitinase I and heparinase of the constitutional unsaturated disaccharide (delta Di-SHS) derived from heparan sulfate (HS) isomers and heparin was achieved using high-performance liquid chromatography (HPLC) with a sulfonated styrene-divinylbenzene copolymer. Eight delta Di-SHS products derived from HS isomers were identified. Enzymatic digestion with heparitinase I and heparinase converts heterogeneous sulfated HS isomers and heparin into different delta Di-SHS. The practical application of these enzymes was examined using specific enzymes and HPLC. In a patient with Hurler syndrome, eight individual delta i-SHS were identified in urinary HS isomers. PMID- 7493083 TI - Protein-boronic acid conjugates and their binding to low-molecular-mass cis-diols and glycated hemoglobin. AB - Different methods for covalent linkage of phenylboronic acid (PBA) to structural proteins and enzymes are presented. Protein-PBA conjugates, free in solution or immobilised on magnetizable polymer particles, were tested for their binding of D sorbitol, D-mannose and glycohemoglobin (GHb). Similarly, alkaline phosphatase PBA conjugates were used in an attempted enzyme-linked sorbent assay for the detection of GHb. Affinity chromatography on immobilised D-mannose and gel chromatographic studies of protein-PBA complexes with [14C]sorbitol, clearly illustrated a low affinity of the interaction studied. Glycated hemoglobin could not be detected using the enzyme-linked sorbent assay approach. However, GHb was found to be specifically retained on columns filled with protein-PBA-coated particles as affinity matrix, enabling the glycation level of blood samples to be determined. PMID- 7493084 TI - Determination of the (R)- and (S)-enantiomers of salsolinol and N methylsalsolinol by use of a chiral high-performance liquid chromatographic column. AB - A new method for the quantitative determination of the enantiomers of salsolinol and N-methylsalsolinol, biologically important alkaloids, is reported. The enantiomers were completely separated without derivatization, using a cyclodextrin-modified silica gel column with an HPLC-electrochemical detection system. The HPLC conditions were examined for the best resolution. The method was sensitive enough to detect salsolinol and N-methylsalsolinol at a concentration of less than 0.1 pmol per injection. In the product of the Pictet-Spengler reaction of acetaldehyde with dopamine or epinine, almost equimolar (R)- and (S) enantiomers of salsolinol and N-methylsalsolinol were detected. Preliminary results indicate that the (R)-enantiomer of both isoquinoline derivatives predominate in the human brain. PMID- 7493085 TI - Particle beam liquid chromatography-mass spectrometry of triphenylmethane dyes: application to confirmation of malachite green in incurred catfish tissue. AB - Eight triphenylmethane dyes (malachite green, leucomalachite green, gentian violet, leucogentian violet, brilliant green, pentamethyl gentian violet, N',N' tetramethyl gentian violet and N',N"-tetramethyl gentian violet) have been characterized by particle beam liquid chromatography-mass spectrometry. The electron ionization spectra obtained of these dyes by this technique exhibit similar fragmentation, with the formation of phenyl and substituted phenyl radicals, and loss of alkyl groups from the amines. It was observed that the six cationic dyes are reduced in the mass spectrometer source to form the corresponding leuco compounds. This technique was evaluated for the confirmation of malachite green and leucomalachite green in incurred catfish (Ictalurus punctatus) muscle tissue. PMID- 7493086 TI - Enriched stable isotopes of elements used as tracers: methods of presenting high performance liquid chromatographic-inductively coupled argon plasma mass spectrometric data. AB - High-performance liquid chromatography (HPLC) of biological fluids and tissue cytosolic preparations was employed in conjunction with argon-induced inductively coupled plasma mass spectrometry (ICP-MS) to investigate the distribution of stable isotopes as tracers. The common way of presenting the data from the ICP-MS is by plotting the count rates versus the retention time of HPLC fractions. Additional information can be derived, e.g., the composite peaks can be further resolved, and the level of enrichment in various biological components can be expressed by alternative ways of presenting these data. The two additional approaches described here involve presenting the ratios of enriched tracer with a suitable naturally abundant mass number of the same element, and by expressing the extent of enrichment by the tracer isotope in a given fraction to that of the same mass number in the fraction derived from an untreated source. Each method of presentation has different merits and drawbacks. The data therefore may be best presented in more than one way to emphasize the conclusions from a given experiment. Observations are presented after simultaneously injecting stable isotopes of three essential elements, copper, selenium and zinc, into mice. Plasma and liver cytosolic fractions were analysed and data represented in different ways as indicated above. PMID- 7493088 TI - Simultaneous determination of moexipril and moexiprilat, its active metabolite, in human plasma by gas chromatography-negative-ion chemical ionization mass spectrometry. AB - A sensitive and selective method for the simultaneous determination of moexipril and moexiprilat, its active metabolite, in human plasma is described using quinapril and quinaprilat as internal standards. The analytes are isolated from human plasma by means of Bond Elut C18 cartridges, methylated with diazomethane, purified by acid-base partitioning and converted to the corresponding trifluoroacetamides using trifluoroacetic anhydride. Moexipril and moexiprilat were analysed by gas chromatography-negative-ion chemical ionization mass spectrometry (GC-NICI-MS) at the fragment ions m/z 302 and m/z 288, respectively. The lower limit of quantitation both for moexipril and moexiprilat is 0.5 ng/ml of human plasma. Linear calibration curves are obtained over the concentration range 0.5-300 ng/ml of human plasma. In any case the imprecision and the inaccuracy are < 15%. The present method has been successfully applied to various pharmacokinetic studies in human subjects and patients. PMID- 7493087 TI - Comparison of reversed-phase and ion-pair chromatography for the determination of strychnine in animal tissues. AB - Ion-pair and reversed-phase high-performance liquid chromatography (HPLC) were evaluated for quantification of strychnine in mountain beaver tissues. Retention time shifts hindered strychnine quantification with both HPLC systems. Co extracted free fatty acids released during storage formed ion-pairs with strychnine, resulting in increased retention by reversed-phase HPLC. Competition with co-extracted basic compounds is likely responsible for the decreased retention of strychnine by ion-pair HPLC. Following an acid-base clean-up, optimal results were obtained with reversed-phase HPLC. Ion-pair chromatography was then used for qualitative confirmation of strychnine residues. PMID- 7493089 TI - Quantification of a dual angiotensin I-converting enzyme-neutral endopeptidase inhibitor and the active thiol metabolite in dog plasma by high-performance liquid chromatography with ultraviolet absorbance detection. AB - MDL 100,240 ([4S-[4 alpha,7 alpha(R*), 12b beta]]-7-[[2- (acetylthio)-1-oxo-3 phenylpropyl]amino]-1,2,3,4,6,7,8,12b-octahyd ro-6-oxo- pyrido[2,1 a][2]benzazepine-4-carboxylic acid, I) is the thioacetyl prodrug of the active thiol, MDL 100,173 (II), a dual inhibitor of angiotensin-I converting enzyme (ACE) and neutral endopeptidase (NEP). A drug which simultaneously inhibits both ACE and NEP may provide a unique therapy for hypertension and congestive heart failure. Methods based on high-performance liquid chromatography with UV absorbance detection at 200 nm were developed to support preclinical pharmacokinetic investigations. One method is used to measure unchanged I and free II, while the second method is used to quantify the total level of the thiol II after the plasma is incubated with the disulfide reducing agent, dithiothreitol. By either method, the analytes are quantified over the range of 25-1000 ng/ml with good accuracy and precision. The overall extraction efficiencies of unchanged I and free II in dog plasma were 79% and 86%, respectively, while the extraction efficiency of total II averaged 75%. Described in this report are the results obtained in validating the assay methods for measuring the compounds in plasma. Pharmacokinetic data are presented which were obtained by applying these methods to plasma collected from dogs dosed with I. PMID- 7493090 TI - MPA--rural bonus. PMID- 7493092 TI - Casemix: your views. PMID- 7493091 TI - Hearts, minds & prostates. PMID- 7493094 TI - Chasing the cents wastes the dollars. PMID- 7493093 TI - Aboriginal and Torres Strait Islander health workers: the unsung health providers. PMID- 7493095 TI - Nursing and the law. Failing to treat childhood epilepsy. PMID- 7493096 TI - Haemochromatosis and raw oysters: a dangerous combination. PMID- 7493097 TI - Hon John Howard MP: leader of the opposition. Interview by John Moran. PMID- 7493099 TI - Nursing on the net. PMID- 7493098 TI - Re-union, re-vitalise, re-cruit, re-present. PMID- 7493100 TI - The Hon Paul Keating: Prime Minister of Australia and leader of the Australian Labor Party. Interview by John Moran. PMID- 7493101 TI - Breast screening: a personal view. PMID- 7493102 TI - Nursing and the law. Consent and trust: pre-op warning signals. PMID- 7493103 TI - HIV/AIDS--no room for complacency. PMID- 7493104 TI - Colors of the spectrum. Orthopedic nursing: a "joint" effort. PMID- 7493105 TI - I'll pass this round. PMID- 7493106 TI - Management perspectives. Our hospital is scheduled to undergo work restructuring. PMID- 7493107 TI - Area hospitals teaching adolescents to become safe sitters. PMID- 7493108 TI - JCAHO ... they're back. PMID- 7493109 TI - Wheelchair etiquette. PMID- 7493111 TI - [Kaposi's sarcoma with ocular location in Zaire]. AB - OBJECTIVE: a clinical study of patients with ocular adnexal Kaposi's sarcoma was undertaken to study its clinical characteristics. METHODS: A review of all patients with biopsy confirmed ocular adnexal Kaposi's sarcoma seen between 1962 and 1991. RESULTS: There were eleven patients during this period with the incidence of 1.25 for 10,000 cases. Ages ranged from one to 47 years (average, 33 years). There were 6 males and 5 females. One (9%) patient was seen during the period of 1961 and 1981 and 10 (91%)during the period 1982 and 1991. Of the 10 patients, 9 had ocular Kaposi's sarcoma lesions related to acquires immunodeficiency syndrome. Lesions involved the right and the left eyes in 50% of cases. Localization of lesions was palpebral in 9 (82%) patients and conjunctival in 2 (18%) patients. The clinical appearance was tumoral, modular, polypoid and as subconjunctival hemorrhage. Three patients underwent excision of tumor. The recurrence was of 100% in approximately three weeks of time. All patients infected with HIV died. CONCLUSION: Ocular Kaposi's sarcoma was not common and its presence is related in most of cases to HIV infection and could be the first manifestation of HIV infection. PMID- 7493110 TI - [Tumor of the maxillary sinus with orbital invasion]. AB - The authors report the case of an epidermoid carcinoma, originating from the maxillary sinus, with orbital extension. The first harmless symptoms can mislead the diagnosis. Other signs will appear a few weeks later leading to a tumoral process. The further examinations will confirm the fears. PMID- 7493112 TI - [Primary orbital lymphoma within the context of Sjogren's syndrome secondary to rheumatoid arthritis]. AB - We are describing the case of a seventy-four year old woman presenting a voluminous hard and painless tumor of the right lids, which has appeared half a year ago. Past medical history consists in a Sjogren's syndrome secondary to rheumatoid arthritis. The right eye is not visible and the left eye presents an important xerophthalmia and an opaque and vascularized cornea. CT Scan and biopsy allow to diagnose a diffuse monocytoid B cell non hodgkin-lymphoma, starting from the lacrymal gland with an extension to the lids and the anterior orbit. The appearance of a non hodgkin lymphoma in the course of the evolution of a rheumatoid arthritis and/or Sjogren's syndrome has been described well. The monocytoid feature of this lymphoma with the Sjogren's syndrome has recently been reported. However, the orbital localisation of this lymphoma is rare. PMID- 7493113 TI - W incision for combined cataract and glaucoma surgery. AB - The W incision can be regarded as a self sealing incision with an additional posterior flap. For cataract surgery, the flap can be used to add further stability to the incision whilst for combined glaucoma surgery it can also be used to "cover" a trabeculectomy located underneath. Up to now, 14 eyes underwent a combined cataract and glaucoma surgery using the W incision. In this article the short term postoperative results and advantages of this new technique in combined cataract and glaucoma surgery are discussed. PMID- 7493114 TI - Clinical and genetical manifestations in 34 families with Leber's hereditary optic neuropathy (LHON). AB - The clinical and genetical manifestations of 64 patients of 34 families with LHON, taken up in an ophthalmogenetic register for LHON, are discussed. The disorder appeared to occur sporadically in 41.2% of the patients. Genotypic characteristics of the pedigrees revealed the 11778 mutation in 57.5% of the families, the 3460 mutation in 21.2% and the 15257 mutation in 9.1%. One family carried only a secondary mutation pattern. Fourteen percent of patients with the 11778 mutation showed recovery of their visual acuity. Visual recovery with the 3460 and 15257 mutation was respectively 21% and 33%. Our youngest patient was 4 years old. PMID- 7493115 TI - Ocular anomalies in an unbalanced translocation between chromosomes 7 and 8. AB - A translocation between chromosomes 7 and 8 was found in several members of a family. We describe two family members with an unbalanced form of this translocation with the derivative of chromosome 7:der(7),t(7;8)(q34;q24.1). Both present multiple congenital systemic anomalies and ocular aberrations. Of those the coloboma of the optic disc is the most obvious. We discuss the pathology of the eyes of one of our patients. PMID- 7493116 TI - Septo-optic dysplasia. AB - The authors present three patients with the diagnosis of septo-optic dysplasia or 'de Morsier' syndrome. They stress the important role of the ophthalmologist in the diagnosis. With the advent of MRI, subtle central nervous system abnormalities have been recognized. A close ophthalmological and endocrinological follow-up is necessary. PMID- 7493117 TI - [Congenital malformations of the eyeball and its appendices in Zaire]. AB - OBJECTIVE: to determine the epidemiology of congenital eye malformations. METHODS: the records of 1740 patients (1913 eyes) with congenital eye malformations seen from 1962 through 1992 were reviewed. RESULTS: the prevalence rate of congenital eye malformations was 2.2%. Congenital cataract (38%), atresia of the naso-lacrimal duct (10%), congenital glaucoma (9%), congenital ptosis (8%), microphthalmos (8%), albinism of iris (8%), microcornea (7%) were the most frequent eye malformations. The common types of associated malformations included albinism, microcephaly, cardiopathy and anomalies of ears. CONCLUSION: the findings of this study were similar to those published in Europe and United States. PMID- 7493118 TI - W incision for cataract surgery. AB - We suggest a modification of the frown type incision for the implantation of rigid intraocular lenses: a W shaped incision. It has improved the control of astigmatism, the self sealing characteristics, needs no special requirement for suture material, has a high mechanical stability, and is easily extendible for ECCE or trabeculectomy. The advantages of the incision are substantiated in a comparative study (invers u and W configuration) in 203 eyes with a 3 months follow up. PMID- 7493120 TI - Transsclerally sulcus fixated anterior chamber lenses in eyes without posterior capsule support: three years follow up. AB - Three years ago we started to implant commercially available anterior chamber lenses with S-shaped haptics an multipoint support into the posterior chamber in cases of secundary lensimplantation after intracapsular cataract operation. The lenses were transsclerally fixed by two sutures through the ciliry sulcus. They exhibit more stability than the C-loop posterior chamber lens and induce some scar formation possibly enabling them to stay firm even after degradation of the suture. PMID- 7493119 TI - Primary posterior capsulorhexis in phacoemulsification: short term results. AB - Twenty-eight eyes were followed for at least 6 months after phacoemulsification, posterior capsulorhexis and intracapsular lensimplantation. No case of clinical significant macular edema, retinal tear or retinal detachment was noted during this period. PMID- 7493122 TI - Cataract-operation with topical anesthesia and i.v. sedation. AB - Topical anesthesia with intravenous sedation was used in 100 cases of phacoemulsification. Results show that this technique is a good and safe alternative for retrobulbar and peribulbar anesthesia. PMID- 7493121 TI - Differentiated treatment of secondary cataract following extracapsular cataract operation. AB - 161 eyes with Nd:YAG laser posterior capsulotomy after in the bag IOL implantation 1-3 years before, were actively followed over a period of 24 +/- 6 months. No CME nor retinal detachment nor induced glaucoma was found. Therefore a YAG capsulotomy in eyes with completely in the bag implanted IOL seems to assure the same level of security than capsular polishing in eyes with sulcus implanted lenses. PMID- 7493123 TI - Extracapsular lens extraction with posterior chamber lens implantation and vitrectomy for diabetic proliferative retinopathy in one operation. AB - Vitreous surgery is often necessary to treat proliferative diabetic retinopathy. Sometimes cataract-extraction is indicated. A series of 37 eyes with ECCE, PC-IOL and vitrectomy in one operation is reviewed. PMID- 7493124 TI - Method for detection of conjunctival chlamydial infection and allergic conjunctivitis. AB - In this study 104 adult patients with chronic conjunctival irritation without obvious diagnosis were investigated by conjunctival scrapings. The smears were stained with May-Grunwald-Giemsa or fluorescein-conjugated monoclonal antibodies prepared against Chlamydiae. In 45 cases it was possible to make a correct diagnosis of Chlamydia infection (29%) or allergy (20%). PMID- 7493125 TI - [A case of keratitis due to Mycobacterium chelonae]. AB - The authors presents one case of primitive corneal mycobacterium keratitis after corneal graft. This rare form of keratitis is analysed in the field of mycobacterial infections who's nowadays more developed in our industrialised countries. The description of the case point out the multiresistance of this mycobacterium and describe the different treatments. PMID- 7493126 TI - Probing and silicone intubation of the lacrimal system in adults. AB - We evaluated an atraumatic catheterisation of the lacrimal system in 23 cases under local anaesthesia in adults. By using this method 60% of the eyes with canaliculus stenosis and about 40% of the eyes with inferior lacrimal duct stenosis could be treated successfully. We suggest that lacrimal duct catheterisation under local anaesthesia can be used as a substitute for DCR in selected cases especially in elderly patients giving a good chance of cure without invasive operation. PMID- 7493128 TI - Ocular findings in prematurely born children at the age of 12. AB - A group of 79 prematurely born children was assessed ophthalmologically at the age of 12 and compared with a group of 100 12 years old children born at term. Of the 79 children, results of neonatal eye examinations were available in 26 prematures (52 eyes). In 28 eyes no signs of retinopathy of prematurity (ROP) were detected (non-ROP group). In 24 eyes a stage 1 or 2 retinopathy of prematurity was present (ROB group). 75% of the children with ROP had an ophthalmological problem. Myopia (29%), hypermetropia (21%), astigmatism (37%) and strabismus (33%) were significantly higher. Considering the whole group of prematurely born children, there was more astigmatism (11%) and strabismus (13%). Preterm children, with or without ROP, are more prone to develop ocular problems. PMID- 7493127 TI - Remote diagnosis in ophthalmology. AB - In cooperation with the C.H.U.-Lille (Eye Department) we set up an interregional and european project. We explain the possibilities and difficulties of remote diagnosis. The aim is to build up a network between different ophthalmological centers and so to give access to different centers of expertise to have a faster and more reliable diagnosis for the benefit of the patient. PMID- 7493129 TI - [Treatment of strabismus in ambulatory surgery]. AB - Since april 1993, we have an out-patient surgery department in our university hospital. We report our experience of out-patient surgery in 80 strabismic patients> We have divided the 80 patients in a pediatric group and an adult group. We describe the conditions of admission for out-patient surgery, the method of anesthesia, the development of the procedure. Surgical techniques do not differ from those used in hospitalized patients. We report our statistics concerning age, diagnosis, type and length of surgery of our 80 patients. The lack of complications and the satisfaction of our patients convince us to carry on out-patient surgery for strabismus. PMID- 7493130 TI - Surgical technique for patients with the apractic type of essential blepharospasm: case report. AB - A frontalis suspension was carried out in a patient with an essential type of blepharospasm, characterized by difficulties in initiating the act of lid elevation, often referred to as the apractic form of blepharospasm or, as J. Elston proposed, the pretarsal blepharospasm. The patient tries to open the eyes by using the frontalis muscle or by manual traction. It is known than in this form of blepharospasm, insufficient results are seen after botulinum toxin infection. Proper examination of the skin crease of the upper eyelid and of the eyelid gives an idea of the insertion of the levator aponeurosis and of the levator muscle function. A desinsertion, due to frequent manual traction, may be found. In this case, reinsertion of the aponeurosis may relieve the symptoms. If no desinsertion is present a frontalis suspension, similar to those used in ptosis surgery, may give good results. PMID- 7493132 TI - [Is treatment with corticotherapy or radiotherapy effective for involvement of ocular motility and for reduction of exophthalmos in Basedow's disease?]. AB - It is acknowledged that high-dose corticotherapy, radiotherapy, or surgical decompression of the orbit are often efficient in dysthyroid optic neuropathy. Corticotherapy and radiotherapy are also given to patients with dysfunction of eye-muscle motility and/or proptosis in the absence of visual loss. The latter indication has been retrospectively evaluated in 13 patients. The limited character of our series is partially explainable by strict inclusion criteria (objective evaluation of muscle dysfunction and proptosis). We failed to demonstrate any significant functional improvement after oral corticotherapy and/or radiotherapy. It thus appears that our study sheds doubt on the actual efficacy of these treatments in patients presenting with not very evolutive involvement of eye-muscle motility. A larger study aimed at comparing the benefits and drawbacks of these treatments is consequently to be encouraged. PMID- 7493131 TI - Treatment of traumatic optic neuropathy, a case report. AB - We report a case of post-traumatic optic neuropathy in which visual improvement, documented with visual evoked potentials (VEP), occurred after administration of a corticoid megadose. The presentation of a single case does not allow to extrapolate conclusions about the efficacy of the steroid treatment, but serves as an illustration for a controversial problem as the management of post traumatic optic neuropathy. PMID- 7493133 TI - Molecular pathology, informed consent, and the paraffin block. PMID- 7493134 TI - Molecular diagnosis of fragile X syndrome. PMID- 7493135 TI - The emerging molecular genetics of sarcoma translocations. AB - Many types of sarcomas are characterized by specific chromosomal translocations which are likely to be of etiologic significance. The recent elucidation of the molecular structure of the several of these translocations has revealed some striking similarities. Nearly all appear to result in the production of novel, tumor-specific chimeric transcription factors. Furthermore, six of the translocations, namely the t(11;22), t(21;22), and t(7;22) of Ewing's sarcoma, the t(12;22) of clear cell sarcoma, the t(12;16) of myxoid liposarcoma, and the t(11;22) of desmoplastic small round cell tumor, achieve this following a peculiar pattern, consisting in the fusion of a gene with an RNA-binding domain (EWS or TLS) with a transcription factor gene (FLI1, ERG, ETV1, ATF-1, CHOP, or WT1). The observation that the different translocation partners of the EWS gene are specifically associated with several distinct types of primitive sarcomas suggests a model in which the translocation partner supplying the DNA-binding domain confers the target specificity of the transcriptional activation mediated by these chimeric proteins, whereas the partner supplying the N-terminal domain and promoter region determines their transactivation potential and expression level. Further analysis of the normal functions and expression patterns of these genes should yield insights into the histogenesis of these different tumor types and into normal tissue development and differentiation. Clinically, our new understanding of the molecular structure of these translocations opens new avenues for molecular diagnosis and investigative therapeutics. PMID- 7493136 TI - Primary cutaneous Ewing's sarcoma/peripheral primitive neuroectodermal tumors in childhood. A molecular, cytogenetic, and immunohistochemical study. AB - Childhood cutaneous and subcutaneous malignancies are rare and include metastatic tumors of diverse histogenesis as well as primary lesions, such as sweat gland carcinomas. Some cutaneous malignancies exhibit a small round cell tumor morphology with few definitive differentiating features; they can thus pose a significant diagnostic problem. We describe two primary malignancies of the skin and superficial subcutis, which were originally diagnosed as sweat gland carcinomas on the basis of their morphological features. A cytogenetic analysis performed on one of these lesions showed the t(11;22)(q24;q12) rearrangement, believed to be unique to the Ewing's sarcoma/peripheral primitive neuroectodermal tumor (ES/pPNET) group of neoplasms. In view of this unexpected result, reverse transcriptase-polymerase chain reaction analysis was performed on both lesions and showed that they expressed EWS/FLI-1 fusion gene mRNA transcripts, the molecular equivalent of t(11;22)(q24;q12). The two tumors also had an immunohistochemical profile suggesting ES/pPNET, including strong expression of the MIC2 antigen. Both patients were treated with wide local excision, and one was given a course of chemotherapy. Neither patient showed evidence of tumor elsewhere after follow-up periods of 2 years and 16 years. These findings suggest that these tumors are indeed a form of primary ES/pPNET arising in the skin or superficial subcutis, which may be of low-grade malignancy and curable by local surgery. PMID- 7493138 TI - Molecular detection of a common mutation in coagulation factor V causing thrombosis via hereditary resistance to activated protein C. AB - More than half of all patients with familial or recurring venous thrombosis have hereditary resistance to activated protein C (HRAPC) as the result of specific missense mutation in the gene for coagulation factor V. Because the mutant factor Va (with an Arg to Gln substitution at codon 506) cannot be cleaved and inactivated by activated protein C, carriers of this mutation are at significantly increased risk of venous thrombosis. We have recently introduced a direct polymerase chain reaction (PCR)-based clinical diagnostic test for the factor V codon 506 mutation based on the destruction of an Mnl I restriction site by the causative nucleotide substitution. To assess the accuracy of this PCR based assay, we compared a functional clotting time test for HRAPC with the direct mutation test. Of 47 patients dually tested, only five had discrepant values for the functional test versus the DNA test. Either of these two complementary assays is useful for the accurate diagnosis of HRAPC. The DNA-based test is, however, specifically recommended for evaluation of anticoagulated patients or patients with borderline functional tests and confirmation of genotype in HRAPC families. In an additional analysis of 287 normal individuals, we found an extremely high prevalence of the mutated codon 506 allele-- approximately 4% in each of two different populations. The absence of disease in the majority of heterozygous carriers suggests that symptomatic thrombosis requires the simultaneous presence of both a mutated factor V protein and additional synergistic factors. PMID- 7493137 TI - Analysis of clonality by polymerase chain reaction for phosphoglycerate kinase-1. Heteroduplex generator. AB - Polymerase chain reaction (PCR) amplification has been used to determine the clonal composition of tissues based on analysis of the pattern of X-chromosome inactivation, but its use has been limited by technical difficulties. This report presents an expedited method to use PCR in the analysis of clonality. The method uses gel electrophoresis of heteroduplexes formed with an artificial heteroduplex generator (HG) and PCR products from the phosphoglycerate kinase-1 (PGK-1) gene from the tissue sections. Amplification was successful in 36 of 37 cases originally diagnosed as endometrial adenocarcinoma. HG analysis of 36 cases confirmed heterozygosity in 12 cases (33.3%). PGK-1 PCR amplification product was obtained from both control and lesional tissue in 10 of the 12 heterozygous cases. Of these 10 cases, seven were shown to consist of clonal cell populations by HG analysis. Two of three cases diagnosed as well-differentiated endometrioid adenocarcinoma were found to be comprised of polyclonal populations of cells. One case produced an anomalous pattern with HG analysis and was shown to be aneuploid by fluorescence in situ hybridization (FISH) with a chromosome X alpha-satellite probe. It is concluded that HG is a useful alternative to restriction fragment length polymorphism (RFLP) analysis of X-chromosome inactivation as a marker of tissue clonality in cases in women. PMID- 7493139 TI - Adenoma-carcinoma sequence of colorectum. Prevalence of K-ras gene mutation in adenomas with increasing degree of dysplasia and aneuploidy. AB - One hundred and fifty colorectal adenomas were investigated in order to detect the presence of K-ras gene mutation. The adenomas were classified according to the severity of the histological lesion (mild, moderate, or severe dysplasia and carcinomatous transformation) and to the degree of aneuploidy. K-ras mutation was found in 30.8% of cases, mostly consisting of a point mutation of codon 12. K-ras mutation was more frequently found in adenomas > 1 cm and in the villous type. No correlation was otherwise demonstrable with the ploidy pattern of the lesion. PMID- 7493140 TI - Alterations of the TP53 gene as a potential prognostic marker in breast carcinomas. Advantages of using constant denaturant gel electrophoresis in mutation detection. AB - Reviewing studies of alterations of the TP53 tumor suppressor gene in human breast carcinomas gives cautious optimism about TP53 alterations as a prognostic marker in this disease. For the time being, breast carcinomas should be screened for TP53 alterations at both the protein and gene levels. Improved mutational screening techniques are needed for this purpose. We consider constant denaturant gel electrophoresis, a modification of denaturing gradient gel electrophoresis, to represent such an improvement. With the recent development of the BioRad D GENE system, constant denaturant gel electrophoresis screening for TP53 mutations can easily be performed on large series of breast carcinomas. PMID- 7493141 TI - Detection of myeloperoxidase gene expression in minimally differentiated acute myelogenous leukemia (AML-M0) using in situ hybridization. AB - Acute leukemias containing > 3% myeloperoxidase (MPO)-positive blast cells, as detected cytochemically, are considered to be myelogenous in origin, regardless of the immunophenotypic markers expressed. Conversely, acute leukemias that express only myeloid antigens are also considered to be acute myelogenous leukemia (AML), even in the absence of MPO. These MPO-negative AMLs, designated AML-M0 in the FAB classification, currently require either immunophenotypic or electron microscopic studies for identification. To examine the association of MPO and myeloid antigen expression in AML, particularly at the early stages of myeloid cell differentiation, we have used in situ hybridization (ISH) to evaluate MPO gene expression in myeloid leukemia cell lines and a variety of well characterized acute leukemias, including six cases of AML-M0. Strong positivity for MPO mRNA was detected in the myeloid leukemia cell line HL-60 and in 22 of 27 AMLs (three AML-M0, four AML-M1, eight AML-M2, five AML-M4, two AML-M5a). No MPO gene expression was detected in three AML-M0, one AML-M5a, one AML-M7, 5 acute lymphoblastic leukemia, the lymphoid cell lines Molt-4 and Namalwa, or in the early myeloid cell lines KG-1 and KG-1a. Ultrastructural studies for MPO activity were performed on four AML-M0; one leukemia showed both gene expression and cytochemical activity, whereas two others contained neither MPO transcripts nor enzyme. Weak MPO gene expression was evident in one AML-M0 that was negative for enzymatic activity by electron microscopy. These studies show MPO gene expression can be detected by ISH in about half of AML-M0, supporting their presumed myelocytic derivation.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7493143 TI - The twenty first century uptake and distribution of anesthetic gases. PMID- 7493142 TI - Estrogen receptor functional status in human breast cancer. AB - Estrogen receptors (ER) are detected in 50-85% of all breast tumors, and are clinically important because they tend to identify patients with a higher probability of responding to hormonal or endocrine manipulations. However, approximately 30-40% of all ER+ patients do not respond to hormonal manipulations. The lack of response to hormonal manipulations in ER+ patients could be the result of nonfunctional ER, as determined by its inability to recognize and bind to specific DNA-responsive elements and/or its inability to recruit other transcriptional activation factors. The functional status of ER in 34 human breast tumors was assessed determining the structural integrity of the ER DNA-binding domain using site-directed monoclonal anti-estrogen receptor antibody and sucrose density gradient analysis. Based on the fraction of ER containing an intact DNA-binding domain, the tumors were classified into three groups: group I with > 65% of intact ER, group II with > 30% of intact ER, group III with < 30% of intact ER. Clinical and pathologic data were obtained only for patients who were treated with the anti-estrogen tamoxifen and correlated with ER functional status. In group I, 11 of 13 (84.6%) patients were responsive to hormonal therapy with favorable clinical outcome; two patients had unfavorable clinical outcome. In group II, 13 of 15 patients (86.7%) had favourable clinical outcome, and two patients 13.3% had unfavorable outcome. In group III, three of six patients appeared to be hormone responsive with favorable clinical outcome, and three of the patients in this group had unfavorable response to therapy.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7493144 TI - A clinical audit for postoperative pain control on 1443 surgical patients. AB - BACKGROUND: Moderate to severe pain occurs in 75 to 100% of patients after major operations. Pain control techniques, for examples the patient controlled analgesia (PCA) and epidural analgesia, are effective in relieving postoperative pain. Routine delivery of these pain control techniques to patients undergoing operations can be provided by an anaesthesiologists-based acute pain service (APS). METHODS: This manuscript audited postoperative pain management service to 1369 ethnic Chinese patients and 74 patients of other races, over a 30 mon period in a teaching hospital in Hong Kong. The APS was prioritized to elderly patient (36.1%, aged 65 or higher), patients of poor physical condition (27.0% ASA III or IV) and those undertaken major operations. Most patients (1348, 93.4%) received either one of the four standardized analgesic regimens including PCA or epidural analgesia. RESULTS: The median verbal quantitative scale of pain (VQS) at rest/movement were 2.0/4.3 on operative day and 0/0.6 on 4th postoperative day. The morphine consumption in 665 Chinese patients (PCA intravenous morphine) was 26.1 +/- 14.3 mg/kg/h (mean +/- SD) during the first 20 postoperative hours. This decreased to 18.1 +/- 15.1 at 21th to 44th and 18.5 +/- 14.9 at 45th to 68th hours postoperatively. There was no difference in morphine consumption between Chinese patients and other races. The incidence for nausea and vomiting were 26.5% and 13.3%, respectively. Respiratory derangement occurred in 23 patients (1.9%) and almost always accompanied by somnolence. Most patients (1176, 81.5%) rated their postoperative analgesia on discharge from the care of APS. CONCLUSIONS: Effective and safe postoperative analgesia in Chinese patients can be achieved with currently available pain control techniques such as epidural and PCA. APS is a practicable approach to deliver these techniques to postoperative patients on a routine basis. PMID- 7493145 TI - The effect of intrathecal bupivacaine with combined fentanyl in cesarean section. AB - BACKGROUND: The use of neuraxial opioids has gained popularity over the last few years; they may augment the analgesia produced by the local anesthetic through direct binding with the specific spinal receptors. Morphine, a lipophobic opioid, may not be optimal as an intrathecal drug for intraoperative analgesia because of its slow onset. The lipophilic opioid, fentanyl for instance, if administered intrathecally, its onset is fast and many of its merits by virtue of its lipophilic property may be seen intraoperatively. METHODS: Seventy five healthy pregnant women who sustained cesarean section under spinal anesthesia were assessed in a randomized fashion. The spinal anesthetic used was 0.5% hyperbaric bupivacaine. Patients were divided into 5 groups, 15 in each group. Fentanyl 0 (Group I), 7.5 (Group II), 10 (Group III), 12.5 (Group IV) and 15 (Group V) micrograms was respectively added to normal saline to make a total volume of 0.3 ml, which was then added to bupivacaine and administered to patients in a randomized fashion. The effect of analgesia, vital signs and side effects were observed every 5 min during operation and every 30 min after operation. RESULTS: It was disclosed that all patients in group V and IV had excellent analgesia intraoperatively, while only 33.3% patients in the control group had an analgesia which was qualified as excellent. Complete analgesia (time from injection to first report of pain) also lasted longer in group IV (201.3 +/- 16.4 min, mean +/ SEM) and group V (210.3 +/- 18.6 min) compared with group I (118.9 +/- 10.4 min). The duration of effective analgesia (time from injection to first parental opioid) was increased with the dose of intrathecal fentanyl above 12.5 micrograms (293 +/- 22.4 min). Both complete analgesia and effective analgesia were not significantly different between group IV and group V. Pruritus was the most common side effect. The incidence of shivering decreased significantly in group IV & V as compared with control group. CONCLUSIONS: The combination of bupivacaine with a dose of fentanyl as low as 7.5 micrograms did not produce actual clinical effects. As the dose of fentanyl was increased to 12.5 micrograms or 15 micrograms the quality of surgical analgesia was better and the postoperative analgesia lasted longer. It seemed that the clinical effect might reach its ceiling at the dose of 12.5 micrograms. Pruritus was the most common side effect, but it was mild. PMID- 7493147 TI - Distance from the skin to the cervical epidural space. AB - BACKGROUND: Cervical epidural blockade provides effective regional analgesia in clinical anesthesia and pain clinics. However, the risk for performing cervical epidural block is higher than lumber epidural anesthesia. We studied the distance from the skin to the cervical epidural space to determine whether there is any relationship between patient age, height, body weight, body mass index (BMI) and the distance from the skin to the epidural space. METHODS: The study included 34 patients who underwent cervical epidural analgesic for orthopedic or plastic surgery. Cervical epidural catheterization was performed at C7-T1 intervertebral space by a midline approach with a 18-G Touhy needle. The depth of the epidural space from the skin was measured by a standard ruler. RESULTS: The distance from the skin to the cervical epidural space at C7-T1 interspace was 4.81 +/- 0.81 cm (mean +/- SD). Linear regression analysis revealed significant correlation between body weight (r2 = 0.53, p < 0.0001), BMI (r2 = 0.58, p < 0.0001) and the depth of the cervical epidural space. CONCLUSIONS: These results indicated that body weight and BMI could be a guideline for identification of epidural space during cervical epidural anesthesia. PMID- 7493146 TI - Magnesium sulfate has negligible effect on middle cerebral artery blood flow velocity in response to endotracheal intubation. AB - BACKGROUND: Pretreatment of magnesium sulfate (MgSO4) attenuates pressor response associated with endotracheal intubation. Vasodilating effect of MgSO4 may increase cerebral blood flow, which upsets the known benefit to cardiovascular hemodynamic. In the present study, we evaluated the effect of MgSO4 on the changes of cerebral blood flow in response to endotracheal intubation. METHODS: Twenty four ASA class I or II patients who randomly received either normal saline 5 ml or MgSO4 (60 mg/kg, i.v.) 3 min before induction of anesthesia were included in the study. Mean blood flow velocity (VMCA) of the middle cerebral artery was obtained using a transcranial Doppler sonography (TCD) before and after endotracheal intubation. Anesthesia was induced with fentanyl (4 micrograms/kg) and thiopental (5 mg/kg) and endotracheal intubation was facilitated with succinylcholine (1.5 mg/kg). RESULTS: Before endotracheal intubation, MgSO4 caused a transient decrease in VMCA (98 +/- 6% vs. 76 +/- 3% of baseline) and increase in heart rate (132% vs. 114% of baseline) compared to saline group. However, VMCA and heart rate did not show significant difference in both groups after tracheal intubation. Also, systolic and diastolic blood pressure did not differ during the study. CONCLUSIONS: Our results suggest that MgSO4 appeared to have negligible effects on cerebral blood flow and hemodynamics in response to tracheal intubation. Its attenuating effect on pressor response induced by endotracheal intubation might be over emphasized. PMID- 7493148 TI - [A proper size of endotracheal tube for infant and young child--a retrospective study in Mackay Memorial Hospital]. AB - BACKGROUND: There are more than 2000 pediatric patients receiving surgery in Mackay Memorial Hospital each year. Most of these surgery were performed under general anesthesia with endotracheal tube; therefore choosing an appropriate size of endotracheal tube becomes an important issue in our daily practice. METHODS: Our principle is to choose an uncuffed Mallinckrodt endotracheal tube with a proper internal diameter (ID), ranging from 2.5 mm to 6.5 mm, which could be suitably and gently inserted into the trachea under full muscle relaxation. The tube would then be immediately removed and replaced with a smaller one if facing obvious resistance during intubation. After intubation, a leak test was applied to ascertain that there was no excessive gas leakage. We reviewed all anesthetic records of elective pediatric surgery in the recent 6 years, and the patients whose age above 8.5 years old and body weight (Wt) above 30 kg were excluded from this study. Using age (6476 cases) and Wt (6406 cases) as our parameters, we analyzed these data according to the distribution of each size of uncuffed endotracheal tube (UCETT) in different age and Wt intervals and compared them with the recommended Western reports. RESULTS: Our results revealed that (1) the UCETT size increases as age or Wt increases; (2) considerable spread of UCETT sizes for different age and Wt intervals and basically represent as normal distribution; (3) for the case of even age equal or above 2 years old (up to 8 years old), the ID of the most frequently used UCETT can be memorized as (18 + age in years) divided by 4 or the outer circumference (OC) of the Mallinckrodt UCETT (in French unit, Fr) = 19 + age in years; and (4) Wt as a parameter for tube size selection was as powerful as age (94.76% vs. 94.65%). CONCLUSIONS: From our results, we concluded that "whatever method of predicting tube size is used, tracheal tubes 0.5 mm larger and smaller should be available at the time of intubation so that the proper size can be chosen when the glottis is visualized." PMID- 7493149 TI - [Uptake of anaesthetic gases and vapours]. PMID- 7493150 TI - [Cytochrome P450-dependent monooxygenase system and anesthetics]. AB - The cytochrome P450-dependent monooxygenases constitute the primary enzyme system responsible for the oxidative metabolism of a variety of xenobiotics and endogenous compounds including drugs, carcinogens, fatty acids and hormones. The monooxygenase system consists of multiple forms of P450 enzymes, NADPH-cytochrome reductase and phospholipids. The level sof P450s and associated monooxygenase activities are subject to be regulated by many environmental, physiological, and pathological factors. Inhalation and intravenous anesthetics are all metabolized through these biotransformation enzymes. The pharmacokinetic properties as well as the toxicity of the anesthetics are closely related to the inducing or inhibitory status of the monooxygenase isozymes. To understand the role of cytochrome P450-monooxygenases in drug metabolism is essential for us to handle the drug-to-drug interactions and adverse effects. PMID- 7493151 TI - The anesthetic management of a patient with streptococcal toxic shock-like syndrome--a case report. AB - Streptococcal toxic shock-like syndrome (TSLS), a severe multisystem disorder with rapid progression and fulminant course, is caused by group A beta-hemolytic streptococcal (GAS) infection. The mortality rate is around 30%. It has been reported in the United States and Europe since the 1980s. The recent recognition of this syndrome is thought to stem from the appearance of more virulent strains of streptococci with a greater tendency to produce potent exotoxins than prior ones. Early diagnosis, treatment with penicillin, radical operative debridement and adjuvant therapy with immunoglobulin and plasma exchange may bear a favourable outcome. A case of TSLS who received surgical management in operating room is presented in this paper. PMID- 7493152 TI - The social meanings behind male sex work: implications for sexual interactions. AB - This qualitative study explores the meanings of the commercial sexual encounter between male sex workers and their clients. The study highlights the various social meanings male sex workers attribute to having sex, their typologies of clients, the psychic contexts of male commercial sex, safer sex interactions, and how these issues inform sexual behaviour. The data shows that the meaning attached to the act of having sex is an important aspect of the way in which participants perceive their partners, conduct themselves during sexual encounters, and engage in safe sex practices. Clients are categorized by sex workers according to their perceptions of 'them', which include 'marrieds', 'easy trade', 'undesirables', 'sugar daddies' and 'heaven trade'. Different types of clients pose alternate levels of risk to the safe sex practices of sex workers. The sex worker's definition of commercial sex as work enables him to separate work and personal sex and define work sex as 'not real sex', in which safe sex practices symbolize both the degree of self that is shared and protective work equipment. It was also found that this sample of sex workers do not negotiate safe sex. Rather they use 'modes of interaction' which direct the encounter either towards safe sex, or they refuse to continue with the transaction. The interactive modes identified are 'natural', 'educative', 'challenge', 'other options' and 'walk-out'. These modes of interaction are effective strategies for ensuring safe sex, and can be used by the broader community to gain partner compliance in safe sex practices. PMID- 7493153 TI - The state and the egalitarian, ecclesiastical and liberal regimes of gender relations. AB - In this paper I attempt to construct a comparative analytical framework for the study of the state as a social relation and the regulation of gender relations. The first part of my analytical framework involves an analysis of the women's movement as an agent of political change. In the second part of my analytical framework, three typologies of regimes of gender relations are developed in order to capture the essential features of gender relations in different countries. These typologies are the egalitarian regime of gender relations, the ecclesiastical regime of gender relations and the liberal regime of gender relations. The final part of the comparative analytical framework involves an analysis of the dynamics of transformation of regimes and transition to new regimes. The main contribution of my typologies is that they analyse, on the one hand, how gender relations are regulated by the state in different parts of the economy (social reproduction included) that are relatively autonomous and identify, on the other hand, how social forces influence the nature of the state as a regulator of gender relations. PMID- 7493154 TI - Islam is the solution: Jordanian Islamists and the dilemma of the 'modern woman'. AB - During the past decade, the issue of gender relations and women's conduct and dress has been occupying an increasingly prominent place in the discourse of Islamist movements. This article attempts to situate Arab Islamists' preoccupation with women within the legacy of colonialism and social transformations relating to gender and class. With regard to Jordan, the author links the urgency of the issue with social transformations at the level of gender and class during recent decades, and points out that the key to understanding the prominence of the 'woman question' in Islamist thinking is the fact that the social groups which comprise the traditional constituency of the Islamists are finally experiencing for themselves the socially disruptive implications of new patterns in women's work, education and visibility. In short, the issue of women's modesty and conduct, which was more abstract as recent as one generation ago (when only upper middle and upper class women were visible in the public domain), acquires concreteness and urgency in the rapidly changing social environment. The article also tries to show how the Islamists' framing of the issue in cultural terms has a primal appeal, especially to those social groups most alienated from the insular world of the westernized elite and in search of 'authentic' ways of living in the modern world. PMID- 7493156 TI - Evaluation of functionality of cochlear outer hair cells in patients with retinitis pigmentosa and in their relatives. AB - Hearing functionality was studied in 36 patients with retinitis pigmentosa (R P) and 29 of their relatives. These patients were defined as having bilateral normal hearing on the basis of tonal-threshold audiometric and acoustic-immittance tests. The transiently evoked otoacoustic emissions (T E O A Es) were studied in these normal hearing patients. T E O A Es represent an extremely sensitive method to study the functionality of the outer hair cells of the organ of Corti. When the values of T E O A E amplitude, intensity, and frequency in R P patients and in their relatives were compared with those in control subjects, they were found to be significantly reduced. The T E O A Es were clearly pathological in 52.8% of patients with R P and in 24.1% of their relatives. During embryologic development, there is one transitory axoneme in the outer hair cells of the organ of Corti; this transitory axoneme is important for the organization of the stereocilia. Axonemes are found in mature hair cells, including photoreceptors. The alteration of cochlear outer hair cells in a high percentage of patients with R P and in some of their relatives corroborates the hypothesis that, in some instances, retinitis pigmentosa may be due to a structural anomaly of the ciliated cells. PMID- 7493155 TI - A point mutation in the RDS-peripherin gene in a Spanish family with central areolar choroidal dystrophy. AB - The RDS-peripherin gene encodes a photoreceptor-specific protein that is localized in the outer segment disc membranes of both rods and cones. We screened a Spanish family with central areolar choroidal dystrophy for mutations in candidate genes. A base substitution was identified in the RDS-peripherin gene of one patient and DNA sequencing revealed a C-to-T transition in codon 172, arginine being substituted by tryptophan. The mutation was also detected in two asymptomatic family members who showed irregular pigmentation in the retinal pigment epithelium (RPE). The phenotype is similar to other macular dystrophies caused by mutation in the RDS-peripherin gene. PMID- 7493157 TI - Uveal melanoma and familial atypical mole and melanoma (FAM-M) syndrome. AB - We conducted this study to determine whether occurrence of primary uveal melanoma in the setting of familial atypical mole and melanoma (F A M-M) syndrome (an autosomal dominant cutaneous preneoplastic syndrome) follows a pattern of a hereditary cancer predisposition syndrome. A retrospective review of 4600 consecutive patients with primary uveal melanoma revealed eight patients with biopsy-proven F A M-M syndrome. The clinical profile of these patients was studied and their kindreds analyzed. In patients with F A M-M syndrome, the uveal melanoma occurred at a relatively young age (mean 40 years; range 10-52 years). The diagnosis of F A M-M syndrome preceded or followed the diagnosis of uveal melanoma by as much as 10 years. None of the patients had an associated nonmelanocytic malignancy. Three of the eight patients had a positive family history of melanoma (cutaneous melanoma (2) and uveal melanoma (1). The authors conclude that the occurrence of primary uveal melanoma in the setting of F A M-M syndrome does not follow a clear pattern of a hereditary cancer predisposition syndrome. PMID- 7493159 TI - A complex chromosome translocation resulting in deletion 11p and associated with uveal colobomata. AB - A boy with a complex chromosome rearrangement of chromosomes 3, 10, and 11 resulting in a deletion of the short arm of chromosome II is presented. The occurrence of uveal coloboma as an isolated congenital malformation might suggest a chromosomal site for this ocular anomaly in proximity to the aniridia locus. PMID- 7493158 TI - Multipoint linkage analysis and homogeneity tests in 15 Dutch X-linked retinitis pigmentosa families. AB - Linkage analysis and homogeneity tests were carried out in 15 Dutch families segregating X-linked retinitis pigmentosa (X L R P). The study included segregation data for eight polymorphic DNA markers from the short arm of the human X chromosome. The results of both multipoint linkage analysis in individual families and heterogeneity analysis support the view that there are only two X L R P loci on the short arm of the human X chromosome, with one locus near the OTC gene and one in the vicinity of DXS255. Furthermore, our data confirm the hypothesis that a tapetal reflex in female carriers can be observed more frequently, if not exclusively, in X L R P families of the R P 3 type. PMID- 7493160 TI - Chinese family with retinitis pigmentosa. PMID- 7493161 TI - Use of lethal body burdens to indicate species differences in susceptibility to narcotic toxicants. AB - Lethal body burdens (LBB) for 1,2- and 1,4-dihalogenated benzenes (F, Cl or Br) are determined in rainbow trout of two age-classes. LBBs range from 0.3 to 2.4 mmol/kg. There are no significant differences between the two tested age-classes of rainbow trout. The rainbow trout data are compared to LBBs for 1,2- and 1,4 difluorobenzene in fathead minnow which range from 2.7 to 3.0 mmol/kg, and to LBBs of dichloro- and dibromobenzenes in guppy and fathead minnow [Sijm et al. 1993] which range from 2.7 to 8.0 mmol/kg. Rainbow trout are more susceptible to dihalogenated benzenes than fathead minnow. The LBB can be used as an instrument to examine the intrinsic toxicity of a chemical to a species, and to indicate the susceptibility of a species. Possible reasons for differences in susceptibility among species are discussed. PMID- 7493162 TI - Uniform System for the Evaluation of Substances. V. ESPE, risk assessment for pesticides. AB - This article is the fifth in a series concerning the Uniform System on the Evaluation of Substances (USES). An integral part of USES is the Evaluation System for PEsticides (ESPE). The systems deals with the current practice in The Netherlands concerning the hazard and risk estimation of substances for which a governmental decision is needed before a substances can be marketed or used in The Netherlands, also taking into account EU-legislation. For pesticides a distinction is made between agricultural and non-agricultural pesticides. Both types of substances can be considered in USES. Surface water contamination of pesticides after application on arable land and lixivation of antifouling paints from boats into the water of a harbour show as an example how certain aspects are handled. PMID- 7493163 TI - A modified SOS-Chromotest procedure to test for genotoxicity and cytotoxicity in sediments directly without extraction. AB - A modified SOS-Chromotest bioassay using a chromogenic pad (pad procedure) was developed to test for genotoxicity in sediments directly without extraction. This test is based on the de novo synthesis of beta-galactosidase enzyme by a genetically-engineered E. coli strain PQ37. In the bioassay, an exponential growth phase antibiotic-containing culture of the test bacterium is introduced into a series of tubes with the first tube containing 0.1 gram of sediment. Serial dilutions are then made and the tubes of sediment plus bacterial culture are incubated at 37 degrees C for four hours, followed by placing a drop of each mixture on a chromogenic pad and additional incubation for 20 hours at 37 degrees C. The solid particulates are then washed off with tap water and positive (genotoxic) activity is noted by the presence of a distinctive blue colour on the pad. The SOS-Chromotest pad procedure may be best used as a relative measure of genotoxicity by comparing results to a reference sample. In addition it can also determine sediment cytotoxicity by comparing samples spiked with a genotoxic standard (i.e., 4-nitroquinoline-N-oxide). Preliminary results suggest that this new bioassay is highly sensitive, consistent and discriminating. PMID- 7493164 TI - Antifungal drugs. PMID- 7493165 TI - Genetics of non-medullary thyroid cancer. PMID- 7493167 TI - Loin pain and haematuria syndrome: a somatoform disorder. AB - Fifteen patients with the loin pain and haematuria syndrome (LPH) were compared with 10 patients with complicated renal stone disease referred to the same tertiary centre and matched for age, sex and duration of illness. LPH patients had a history of three times more medically unexplained somatic symptoms other than loin pain (p < 0.01) and a higher proportion took analgesics regularly (p < 0.01). The onset of pain was associated with an adverse psychologically important life-event in eight of the LPH patients but in none of the controls (p < 0.02). LPH patients more frequently recalled serious parental illness and disability in childhood (p < 0.001) than controls, and a higher proportion felt responsible for causing or alleviating parental illness or distress (p < 0.05). LPH subjects scored higher in the 'paternal care' dimension of the Parental Bonding Instrument (p < 0.05). No difference was found between LPH patients and controls in terms of current depression and anxiety but both groups exhibited high rates of lifetime depression. LPH patients expressed lower levels of anger and hostility (p < 0.002) than did controls. Our observations suggest that psychological factors are of major importance in the aetiology of LPH, which may represent a type of somatoform pain disorder. PMID- 7493168 TI - Beeturia and colonic oxalic acid. AB - Beeturia is the excretion of red beetroot pigment (betalaine) in urine and faeces. It occurs in about 14% of humans. Betalaine is a redox indicator whose colour is protected by reducing agents. We investigated pigment-decolourizing systems in the intestinal tracts of beeturic and non-beeturic subjects. Betalaine was decolourized by hydrochloric acid, ferric ions and colonic bacteria preparations, but not by pancreatic or mucosal enzymes. In animals, oral betalaine did not produce beeturia, but injection of betalaine into the peritoneum did. Oral betalaine and 1 g oxalic acid produced beeturia in non beeturic normal humans, but passed into ileostomies without beeturia. Thus, beeturia results from colonic absorption of betalaine. Oxalic acid preserves the red colour to the colon, otherwise it is decolourized in non-beeturic individuals by non-enzymic processes in the stomach and colon. PMID- 7493166 TI - A new apolipoprotein Al variant, Trp50Arg, causes hereditary amyloidosis. AB - A man with hereditary non-neuropathic systemic amyloidosis had amyloid fibril protein subunits consisting of N-terminal fragments (residues 1-86, 1-92 and 1 93) of a previously unknown variant of apolipoprotein Al, Trp50Arg, encoded by a thymine-cytosine transition. This is the third reported amyloidogenic apoAl variant. All involve substitutions of single neutral amino acids by the cationic residue arginine, suggesting a common mechanism of amyloidogenesis. However, the phenotypic expression of these mutations varies both within and between the seven known families with hereditary apoAl amyloidosis. These findings should facilitate analysis of the molecular basis of fibrillogenesis and of factors that modulate amyloid deposition and its consequences in vivo. PMID- 7493169 TI - Octreotide reduces alcohol-induced hypotension and orthostatic symptoms in primary autonomic failure. AB - Symptomatic postural hypotension is a major problem in patients with primary idiopathic autonomic failure, and ingestion of small quantities of alcohol may worsen the degree of postural hypotension. The proposed mechanisms include mesenteric vasodilation and release of vasodilatory gut peptides. We measured systemic, mesenteric, other regional vascular and biochemical responses to alcohol ingestion before and after pre-treatment with the somatostatin analogue Octreotide (which inhibits the release of a wide range of gut peptides normally released in response to food ingestion) in six patients with primary autonomic failure. Octreotide effectively prevented alcohol-induced hypotension and vasodilatation of the mesenteric vascular bed, with improvement of signs and symptoms of hypotension post-alcohol. This suggests that the mechanism of alcohol induced symptomatic hypotension in autonomic failure is at least partly mediated by release of vasodilatatory gut peptides with resultant mesenteric vasodilatation. PMID- 7493170 TI - A pilot study of streptokinase for acute cerebral infarction. AB - We evaluated intravenous streptokinase in the treatment of cerebral infarction. Following neurological assessment and cerebral computed tomography (CT), patients aged 40-80 years with symptoms of anterior circulation acute ischaemic stroke were given 1.5 M units streptokinase or saline placebo in a double-blind randomized study. Twenty patients (10 streptokinase, 10 placebo), 11 males, 9 females, aged 57-79 years, were treated out of 512 consecutive admissions to the acute stroke unit over a 2-year period. Initial CT was normal in 11 (6 placebo, 5 streptokinase) and showed early signs of cerebral infarction in nine (4 placebo, 5 streptokinase). Median times from symptom onset to treatment were 5.2 h (placebo) and 5.8 h (streptokinase). Streptokinase treatment was associated with symptomatic hypotension in one patient. Repeat CT at 72 h demonstrated intracerebral haematoma in two patients and haemorrhagic infarction in one patient in the streptokinase group; the two cases of haematoma formation were associated with neurological deterioration and death. One patient in the placebo group had evidence of haemorrhagic infarction at 72 h. There were three deaths in each treatment group, all within the first 14 days. Patients with acute stroke can be evaluated with CT and treated with streptokinase within 6 h, but the opportunity for treatment is currently limited to few patients. Streptokinase treatment is not without risk, but potential clinical benefit justifies ongoing multicentre randomized trials. PMID- 7493171 TI - Accident experience and notification rates in people with recent seizures, epilepsy or undiagnosed episodes of loss of consciousness. AB - Collaborating neurologists provided a central office with clinical information, driving experience and accident rates in all drivers they counselled after diagnosing seizures or other unexplained episodes of loss of consciousness. Each patient received and was asked to return a notification slip to DVLA. Of 638 patients counselled, 11% had been involved as a driver in a road traffic accident in the previous year. Six patients had been involved in an accident producing serious injury. These estimates do not differ from those expected in a population of non-epileptic drivers with the same age and sex distribution. Of patients counselled, 27.1% returned a reply slip to the DVLA. Notification was more likely if the patient was aged 50 or over at the time of counselling, if the counselling was undertaken by a consultant, if an accident had occurred in the previous year and if the subject had also been advised to start or was already taking anti epileptic drugs. Patients counselled after a single seizure were more likely to notify than those with a diagnosis of epilepsy. PMID- 7493172 TI - An appraisal of Thomas Lewis's effort syndrome. PMID- 7493173 TI - Stories about living in a hospital ward as narrated by schizophrenic patients. AB - Ten schizophrenic patients were interviewed about their own experiences of life in a hospital ward, of their care provider and of the care received. The interview texts were analysed using a phenomenological hermeneutic method. Fatigue, thinking about problems/inner activities, relations and longings for the pre-illness days were themes that occurred in the stories. Seven of the ten interviewees stated that they suffered from fatigue. They were preoccupied with an inner dialogue, trying to solve problems on their own. The patients were alone in their stories. The problems that the patients expressed with regard to relationships arose because they both longed for and feared contact with others, and they felt that they were not 'themselves' when they were with others. For developing care, the importance of learning more about the patients' experiences, especially regarding fatigue, is emphasized. PMID- 7493174 TI - Schizophrenic patients' image of their carers and the carers' image of their patients: an interview study. AB - In order to gain a deeper understanding of how long-term adult schizophrenic patients perceive themselves, their carers and the care they receive in relation to how the carers perceive their patients, themselves and the care they provide, a narrative interview study, analysed using a method inspired by Ricoeur's philosophy, was performed. The main findings of this study were that both patients and carers who saw the patients as a subject stressed the confirming relation between the parties as an essential element of the care. The carers who saw the patient as an object did not share this opinion. They had a more restricted point of view, seeing their goal only as providing a variety of care activities and keeping the schizophrenic patients at the hospital. This study also points out that confirmation is provided in the meeting between patient and carers. PMID- 7493175 TI - Measuring constructs accurately: a prerequisite to theory testing. AB - Accurate identification and measurement of constructs is particularly relevant to mental health nurses, who often have to work with clients that have intangible needs and problems. In this paper, the measurement of constructs, the relevance of construct validity to nursing research and, in particular, to theory testing will be discussed. Constructs within the health belief model are used to illustrate the discussion and procedure of factor analysis. Analysis of data based on a survey of health beliefs of 276 adults with the chronic condition of insulin-dependent diabetes is presented, interpreted and discussed. Whilst the paper focuses upon specific constructs and study population in order to provide a worked example, the issues raised and principles outlined relate to an analytical model that can be applied to other research problems and settings. PMID- 7493176 TI - Research design and methodology for a local mental health needs assessment. AB - The development of a methodology for the assessment of the mental health needs of a local population is described. A framework of corporate consultation was adopted to include methods of accessing the views of key stakeholders. This involved a provider perspective from a survey of clinicians, a user perspective gained through a series of focus groups and a purchaser perspective obtained from semi-structured interview and focus groups. The method adopted for the focus groups was based on an 'ideal research and change practice', which offers potential for development and usage by service planners, including mental health nurses. This article explores issues around qualitative research and the implications for mental health nurses. PMID- 7493177 TI - The phenomenon of clinical supervision in psychiatric health care. AB - The main objective of this study is to describe how nurses experience clinical supervision in psychiatric care. The study design is explorative and descriptive. A clinical supervision process with a theoretical approach to caring includes a learning process, the application of a caring process, and a relationship process. An important aspect of this theoretical approach is the spiritual dimension of caring and its manifestation in the supervision process as well as in the development of the nurse's identity. The results indicate the following phenomena: guilt and a feeling of guilt, reconciliation, forgiveness and loneliness. PMID- 7493179 TI - Measuring patients' perceptions of psychiatric nursing actions. PMID- 7493178 TI - Supportive clinical supervision: a cruicible for personal and professional change. AB - The purpose of this paper is to explore theoretical and practical issues pertinent to the implementation and development of supportive clinical supervision. The paper highlights a facilitative model of supervision that emphasizes the importance of the supervisory relationship within a person-valuing ethos. The authors put a case for sensitive utilization of a supervision concept that leads, they conclude, to self development, self realization, professional enhancement and advancement, and high quality client care. Over time, the ultimate goals of such supervisory practice are twofold: (i) to improve the overall health of the nation and (ii) to promote the progression of the nursing profession. The authors found that a facilitative approach to clinical supervision is therapeutic and self-propelling for both supervisor and supervisee. PMID- 7493180 TI - People, settings and opportunities: an employment feasibility study. PMID- 7493181 TI - The Royal College of Nursing Scottish Board's involvement in the area of mental health. PMID- 7493182 TI - Dementia care mapping. PMID- 7493183 TI - Clinical audit regarding dementia. PMID- 7493184 TI - What to do with nursing models. PMID- 7493187 TI - Development and testing of a measure of depressive cognitions in older adults. AB - Assessment of cognitive processes that predispose or contribute to clinical depression in elders is important for planning interventions that facilitate positive cognitive appraisal and effective coping strategies. Instruments evaluating severity of depression exist; however, none measure specific depressive cognitions. This paper describes the development and pilot testing of the Depressive Cognitions Scale (DCS) for older adults. Derived from Erikson's Psychosocial Theory, the 8 items comprising the scale were selected by clinical experts from a pool of 24 items. Each item reflects a depressive cognition that may arise from less than successful resolution of one of Erikson's developmental phases. Psychometric testing of the DCS was conducted with 60 functionally independent, community dwelling older adults (age 65+). An alpha of .78 indicated acceptable internal consistency. Construct validity was demonstrated by significant correlation with measures of psychosocial development. Further psychometric study of the DCS is recommended. PMID- 7493185 TI - A physiologic method for monitoring premature infants. AB - Instrumentation capable of handling 12 continuous hours of nine-channel real-time physiologic data sampled at 10Hz was needed to test within and between subject variability and preterm infant responses to skin-to-skin contact with the mother. A review of basic electrical components, electrical principles related to physiologic monitoring, and electrophysiology concepts generic to physiologic monitoring is presented. The development, specifications and applications of a new instrument to monitor premature infant cardiorespiratory adaptations are discussed. PMID- 7493186 TI - Using health care records as sources of data for research. AB - Health care records are comprehensive in nature, and provide continuity of health care; therefore, they are vital components in the delivery of services. Health care records also are extremely important for researchers, since they are a rich source of critical information, and the documentation in them is considered to be legally and medically accurate and reliable. This paper examines the advantages and disadvantages of using health care records as data sources for research and discusses the research method issues related to these data sources. The issues addressed are illustrated with examples from three large studies in which health care record reviews represented the only source of data or were a primary information source. PMID- 7493188 TI - Cognitive assessment in the acute care setting: reliability and validity of the Levels of Cognitive Functioning Assessment Scale (LOCFAS). AB - Nurses have responsibility for accurate assessment of cognitive functioning for traumatic brain injured (TBI) patients early in the posttrauma period to enhance rehabilitation with appropriate care planning. To meet this need the Levels of Cognitive Functioning Assessment Scale (LOCFAS) was adapted from the Rancho Los Amigo Levels of Cognitive Functioning instrument. One validity and three reliability studies were conducted using videotapes of five TBI patients at different levels of cognitive functioning. Interrater and intrarater reliability was assessed using coefficient kappa. With raters experienced in assessing cognitive functioning, agreement among cognitive levels was 1.00 with a mean agreement of .997 (SD = .006) for individual items. With inexperienced raters in cognitive assessment, mean agreement for levels was .839 (SD = .120) and for individual items, .830 (SD = .052) Intrarater reliability, after a two-week interval, yielded a mean agreement of .860 (SD = .088) for levels. Criterion related validity of the LOCFAS was demonstrated by correlating it with the Rancho Los Amigos Levels of Cognitive Functioning scale. The resultant Pearson r correlation was .929. Implications for nursing practice and nursing education are discussed. PMID- 7493189 TI - Patient satisfaction with nursing care: concept clarification. AB - Concept clarity is essential for instrument development and validity of research findings. The process of concept clarification can help rectify some of the issues in the measurement of patient satisfaction. This study presents a strategy, for concept clarification, for the purpose of refinement of an existing instrument for measuring patient satisfaction with nursing care in the hospital setting. The results of the exercise yielded a definition of patient satisfaction with nursing care, including a new framework, and provided guidance for rewarding two items, elimination of some items related to knowledge and skill of nurses, as well as generating new items not previously included in the original instrument. PMID- 7493191 TI - Cost effectiveness and AIDS: science or marketing? Proceedings of a workshop. Washington, D.C., January 28, 1995. PMID- 7493190 TI - Reliability and validity of a new preadmission acuity tool for long-term care. AB - The purpose of this project was to develop a new tool designed to assess the acuity of applicants to long-term facilities. Using a known groups method, residents of long-term care facilities were categorized as independent (n = 68), assist (n = 71), and dependent (n = 65) and assessed using the Preadmission Acuity Inquiry Tool (PAI) to generate the data base for examining its psychometric properties. The progression of the mean scores from independent to assist to dependent demonstrates the ability of the tool to differentiate across varying levels of functional independence. Using discriminant analysis, the degree of accuracy in predicting group membership for the independent group was 88%, for the dependent group 75%, and for the assist group 17% with the overall agreement between PAI scores and a priori classifications at 60%. Kappa coefficients for interrater reliability ranged from .54 to 1.0 with a mean of .90. The Cronbach's alpha coefficient was .81. The PAI has demonstrated sufficient reliability and validity to warrant further refinement and testing. This new instrument may enable health care professionals to provide the appropriate level of care in a cost-effective manner. PMID- 7493192 TI - Evaluating the cost-effectiveness of pharmacologic therapy: where we've been and where we're going. PMID- 7493193 TI - Cost-effectiveness: from whose perspective? AB - Evaluating the cost-effectiveness of medical interventions, particularly pharmaceuticals, has been the focus of much debate in this country over the past few years. Although the support for efficient utilization of monies is widespread, the delicate balance between optimizing patient outcome with the costs to do so in high-risk patients is especially difficult. Although research in the field of cost-effectiveness is widespread, at present there are few guidelines in this country. Furthermore, the introduction of bias is a serious concern in the interpretation and validation of studies, often limiting their utility. Guidelines that can be implemented to minimize bias in economic research have been identified to include administrative, methodology-based, and ethical parameters. The standardization and implementation of such guidelines can reduce bias in economic research and optimize study validity. This review focuses on these issues and suggests that initial steps to produce unbiased economic research include total disclosure, appropriate comparator selection, complete release of all relevant data, and the cooperation of all involved parties. PMID- 7493194 TI - Working toward a common currency: is standardization of cost-effectiveness analysis possible? AB - The field of cost-effectiveness has been quietly evolving for several decades. However, concerns and controversies have recently surfaced regarding the lack of standardization with which cost-effectiveness analyses are performed. This attention is due to the increasing use of the technique in rationing health care, setting price and/or reimbursement levels, adopting new technologies, and marketing pharmaceutical products. Is standardization possible while the field is still developing? This article explores possible answers to this question and reviews worldwide efforts aimed at the development of standardized guidelines for cost-effectiveness analysis. PMID- 7493195 TI - Specific considerations for cost-effectiveness studies in AIDS. AB - HIV disease is an epidemic with unique features that make it a challenging topic for pharmacoeconomic analysis. Unlike many other illnesses, the lengthy course of HIV disease, its relatively high incidence among young and productive individuals, and management of drug toxicities have an enormous impact on total costs. Methodologic difficulties discussed include strategies for handling the effect of having a few high-cost cases and for summarizing effectiveness. The advantages and disadvantages of different measures of effectiveness, such as health status, quality of life, and utility in the setting of HIV disease, as well as the scaling of measures and approaches to handling death, are also addressed. PMID- 7493196 TI - Design and assessment of cost-effectiveness studies in AIDS populations. AB - Cost-effectiveness (CE) ratios are indicators of comparative efficiency of competing drugs. Unfortunately, they are subject to variations, depending on factors such as the study population chosen, the comparison treatment selected, and the economic analysis methods employed. CE ratios tend to be unfavorably skewed in the case of chronic illnesses, such as HIV disease, because management of AIDS patients is costly and complex. Routine AIDS therapy requires expensive drugs, treatment is directed at many disease stages and severities, and gain in patient survival is relatively short. Variations in disease stage, co morbidities, and antiviral drug use produce variations in both the risk of opportunistic infections (OIs) and the routine costs of OI prophylaxis and thus affect the CE ratio. Routine therapy for AIDS varies among and even within communities. Overall, these factors translate to a widening gap between trial results (efficacy) and community outcomes (effectiveness). This article discusses these concerns and reviews methodologic issues that must be considered by decision-makers using CE analyses to describe a therapy's value. PMID- 7493197 TI - Understanding the essentials of economic evaluation. AB - Economic evaluation (EE) answers the following simple question: "From which course of action do we get the most value for our money?" We ask this question because resources are always limited, i.e., we never have enough money to do all the things we would like to do. Three types of economic evaluations are used: cost-effectiveness analysis, cost-utility analysis, and cost-benefit analysis. Although all involve a monetary and outcome comparison of two or more courses of action, the methodologies and outcomes of each type vary, making each one particularly suited for specific and different indications. Although the performance of an EE may be complex, its concept is intuitively simple. Understanding the basic elements of economic analysis is more and more important to all health-care providers because health-care policy makers at all levels are increasingly using EE for allocating resources. PMID- 7493198 TI - Workshop on sarcopenia: muscle atrophy in old age. Airlie, Virginia, September 19 21, 1994. Proceedings. PMID- 7493199 TI - The significance of sarcopenia in old age. AB - Our knowledge of the significance of sarcopenia in old age is limited by a lack of epidemiologic data, an incomplete assessment of the pathophysiologic consequences of age-related decrements in muscle mass and quality, and poor understanding of the underlying mechanisms responsible for these decrements. These gaps prevent us from fully appreciating the extent of the public health burden that sarcopenia poses, and present major obstacles to the elucidation of therapies to prevent or reverse sarcopenia in the elderly. The National Institute on Aging convened the Workshop on Sarcopenia to address these issues. The primary questions explored at the workshop included: (a) What research is needed to determine the clinical and functional significance of sarcopenia? (b) What research is needed to understand its etiology? and (c) What opportunities are there for determining the efficacy of current or potential interventions to prevent or retard the development of sarcopenia? The research recommendations from the workshop underscored the need for more dialogue between researchers in different fields (e.g., endocrinology, exercise physiology, bone biology) and for multidisciplinary approaches in order to gain greater insight into sarcopenia. The summary of the research directions which follows has been organized according to the primary questions of the workshop. PMID- 7493200 TI - Effects of aging on energy requirements and the control of food intake in men. AB - Inadequate energy intake can be an important contributor to sarcopenia in older individuals, because negative energy balance induces negative nitrogen balance, independent of nitrogen intake. This review highlights recent studies from our laboratory on possible causes of negative energy balance in older individuals. Studies of energy requirements determined by measurement of total energy expenditure during weight maintenance, using the doubly labeled water technique, and the control of food intake following overeating and undereating indicate (a) that older individuals have higher energy requirements than anticipated, based on current Recommended Dietary Allowances for energy, and (b) that aging is associated with a significant impairment in the control of food intake, which inhibits appropriate compensation for prior changes in energy intake. The combination of unexpectedly high energy needs, and an inability to accurately control energy intake, increases the risk for negative energy balance and associated body fat and protein losses in older individuals. PMID- 7493201 TI - Muscle protein turnover: methodological issues and the effect of aging. AB - Investigation of sarcopenia should include the measurement of muscle protein synthesis and breakdown because reduced muscle mass occurs only when muscle protein content is reduced. Protein content of a tissue is determined by the balance between synthesis and breakdown. Whole body measurements are inappropriate for determining the small changes in muscle protein synthesis and breakdown, since muscle contributes less than 30 percent to the whole body protein turnover. The measurement of mixed muscle protein synthesis in humans supports the animal data, which demonstrates that there is a decline in muscle protein synthesis with age. Resistance exercise stimulates mixed muscle protein synthesis rate in both the young and the old, although this is not demonstrated when myofibrillar protein synthesis is measured. Muscle is composed of several types of proteins, and their turnover may be differentially regulated. It is important, therefore, to measure the turnover of specific proteins such as myosin and actin, which are important contractile proteins. Recently, techniques have been developed to measure the synthesis rate of myosin heavy chain (MHC) in humans. In addition, the application of quantitative polymerase chain reaction (PCR) allows measurement of the steady state mRNA levels of various isoforms of MHC. These measurements, in combination with measurement of the synthesis rate of MHC, will allow us to understand the molecular regulation of specific proteins such as myosin. The important question is to determine whether the decline in muscle protein synthesis is genetically determined or secondary to inactivity, nutritional, hormonal, or other age-related alterations in body functions, and whether muscle wasting is reversible.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7493202 TI - Human aging, muscle mass, and fiber type composition. AB - To assess the age-related loss of muscle mass and to determine the mechanisms behind this aging atrophy, the muscle structure and fiber type composition have been estimated, using invasive and noninvasive techniques. Limb muscles from older men and women are 25-35% smaller and have significantly more fat and connective tissue than limb muscles from younger individuals. Comparisons of muscle biopsies from younger and older individuals reveal that type 2 (fast twitch) fibers are smaller in the old, while the size of type 1 (slow-twitch) fibers is much less affected. Studies of whole muscle cross sections also show a significantly smaller number of muscle fibers, a significantly lower relative type 2 fiber area, and a significant increase in fiber type grouping with increasing age. These results indicate a gradual decrease in size/volume with advancing age, accompanied by a replacement by fat and connective tissue. This aging atrophy seems to be due to a reduction in both number and size of muscle fibers, mainly of type 2, and is to some extent caused by a slowly progressive neurogenic process. PMID- 7493204 TI - Cytokines in aging and muscle homeostasis. AB - Cytokines are best known as mediators of host defense responses to infection and other environmental stresses. However, some of these proteins (interleukin-1 and tumor necrosis factor) may modulate clearance and repair processes in skeletal muscle following injury, and others (fibroblast growth factor and platelet derived growth factor) may be involved with the sustained viability of muscle cells. Muscle repair and vitality also require neuronal contact (influenced by nerve growth factor and ciliary neurotrophic factor) as well as angiogenesis and connective tissue matrix formation (influenced by transforming growth factor beta). Successful muscle aging will depend in part on how well a muscle repairs itself after damage. This includes not only overt injury, but also the daily "wear and tear" that may not be perceived via pain or alterations in function. Age-related loss of muscle mass or function may be the cumulative result of repeated episodes of incomplete repair. Abnormal production or sensitivity to cytokines by aged cells may contribute to these changes in muscle mass and function. PMID- 7493203 TI - Strength and aerobic training attenuate muscle wasting and improve resistance to the development of disability with aging. AB - By the age of 50 yrs old, humans become aware that they are losing muscle strength (mass) and endurance (mitochondria). A frequent symptom of neuromuscular disorders is muscle weakness (Walton, 1988). We define the aging-associated muscle wasting as a progressive neuromuscular syndrome that will lower the quality of life in the elderly by (1) decreasing the ability to lift loads (progressing to difficulty arising from a chair), and (2) decreasing endurance (leading to an inability to perform the activities of daily living, which increases health care costs). Campion (1994) states that the most successful outcome would be for the very elderly to take control of the last stage of their life and make it worth living. To obtain this goal, prevention of muscle wasting is an absolute requirement. Muscle mass and motor unit number, activation, and synchronization are highly related to strength; both decrease with aging (Rodgers and Evans, 1993). Resistance-training is the best way to increase muscle mass, neural coordination, and strength. Mitochondrial concentration is highly related to endurance capacity in young and old (Holloszy and Coyle, 1984). Both muscle contractile and mitochondrial protein decrease with aging in sedentary humans (reviewed by Rodgers and Evans, 1993). Endurance training, which is the best exercise to increase/maintain mitochondrial concentration with aging, has generally resulted in relatively small functional benefits to nursing home patients (Fiatarone et al., 1994).(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7493205 TI - Muscle atrophy and weakness with aging: contraction-induced injury as an underlying mechanism. AB - In old compared with young animals, muscle mass is decreased by 30% to 40%, and maximum force and power are decreased to an even greater extent. The age-related declines in muscle mass and muscle function are similar to those that occur with decreased physical activity. Despite the similarities, we conclude that the losses in muscle mass, force, and power are not due solely to old animals being less active, but rather accrue from intrinsic age-related changes in muscles and in muscle fibers that appear to be immutable and irreversible. The intrinsic changes are associated with denervation of fast fatigable fibers and motor units and motor unit remodeling, which may be initiated by contraction-induced injury. The mechanisms remain unresolved for the weakness, the fatigability, the high susceptibility to contraction-induced injury, and the impaired recovery from injury demonstrated by the skeletal muscles of old animals. PMID- 7493206 TI - The application of blood flow measurements to the study of aging muscle. AB - Blood flow to skeletal muscle is a potentially important factor in the reduction of muscle function associated with aging (sarcopenia). The main influence of reduced blood flow capacity on muscle function is in limiting oxidative metabolism. Direct measures of blood flow include: intravital-microscopy, plethysmography, radioactive microspheres, 133Xenon washout, thermodilution, and Doppler ultrasound. Indirect measurement of blood flow includes arm-to-ankle pressure index and the rate of phosphocreatine recovery after exercise. Several new methodologies have been developed to evaluate muscle blood flow, including color-Doppler imaging, magnetic resonance imaging/angiography (MRI/MRA), and near infrared spectroscopy (NIRS). As adaptations of traditional techniques, these methods promise more precise information under less invasive conditions. MRI is an expensive and technically challenging method able to measure vessel location, blood flow, and wall diameter in blood vessels throughout the cardiac cycle. Color-Doppler provides excellent temporal resolution blood flow throughout the cardiac cycle, along with some anatomical information. NIRS is an inexpensive and portable technology that can measure changes in oxygen saturation and provide information on tissue oxygen delivery in studies of frailer and more difficult-to study subjects. Muscle blood flow is not thought to limit oxidative metabolism under normal conditions in young individuals. However, it is not clear what happens to muscle blood flow in healthy older individuals. Reduced capillary density, less maximal blood flow, and a slower hyperemic flow response have been reported in some, but not all, studies. Further studies with the newer methodologies are needed to re-examine age-related changes in muscle blood flow.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7493207 TI - What insights into age-related changes in skeletal muscle are provided by animal models? AB - This review addresses the following questions: (a) How do the age-related changes in muscle of animal models compare to those found in human muscle?, and (b) How do the changes characteristic of models for reduced or altered muscle use compare to the age-related changes in muscle? Regarding the first question, the vast majority of relevant animal research has focused on the rodents, and the age related changes in rodent muscle are in many respects reminiscent of human aging. A possible difference is the existence of a more profound hypoplasia in human muscle. Research into the age-related changes in muscle in other species is extremely scarce. Regarding the second question, various experimental conditions of reduced or altered muscle use (e.g., denervation, limb immobilization, hindlimb suspension) offer valuable insight not because they exactly mimic normal aging; rather, they demonstrate the multifactorial nature of processes leading to muscle atrophy. PMID- 7493208 TI - Cell culture systems as tools for studying age-related changes in skeletal muscle. AB - Two aspects of muscle cell phenotype that may be affected by aging are amenable to study in vitro: proliferative potential and ability to differentiate. These processes in primary myoblast cultures as well as in established myoblast cell lines will be examined, and the usefulness of these in vitro systems in addressing fundamental molecular mechanisms of muscle aging will be discussed. PMID- 7493209 TI - Effects of exercise on body composition and functional capacity of the elderly. AB - The capacity of older men and women to adapt to regularly performed exercise has been demonstrated by many laboratories. Aerobic exercise results in improvements in functional capacity and reduced risk of developing type II diabetes in the elderly. High-intensity resistance training (above 60% of the 1 repetition maximum) has been demonstrated to cause large increases in strength in the elderly. In addition, resistance training results in significant increases in muscle size in elderly men and women. Resistance training has also been shown to significantly increase energy requirements and insulin action of the elderly. We have recently demonstrated that resistance training has a positive effect on multiple risk factors for osteoporotic fractures in previously sedentary post menopausal women. Since the sedentary lifestyle of a long-term care facility may exacerbate losses of muscle function, we have applied this same training program to frail, institutionalized elderly men and women. In a population of 100 nursing home residents, a randomly assigned high-intensity strength training program resulted in significant gains in strength and functional status. In addition, spontaneous activity, measured by activity monitors, increased significantly in those participating in the exercise program, while there was no change in the sedentary control group. The fact that significant muscle hypertrophy was seen in this population indicates that age does not decrease the capacity to adapt to a progressive resistance training program. Therefore, exercise may minimize or reverse the syndrome of physical frailty which is so prevalent among the oldest old.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7493210 TI - Trophic factor supplementation: effect on the age-associated changes in body composition. AB - Many of the changes in body composition associated with aging are similar to those found in states of growth hormone deficiency. Elderly people have also been found to have significant abnormalities in several trophic factor systems including growth hormone/insulin-like growth factors, and both male and female sex hormones. This has led to intense interest in the clinical effects of trophic factor supplementation in older individuals. This article reviews the age-related alterations in the growth hormone and sex steroid axes, as well as the effects of supplementation. In addition, important questions about the use of trophic factors in the elderly that must be addressed in future research are put forth. PMID- 7493211 TI - Interventions based on the possibility that oxidative stress contributes to sarcopenia. AB - Skeletal muscle is a tissue which accounts for a large part of the body's total oxygen consumption at rest, due to its large mass and the majority of oxygen consumption during vigorous physical activity. Also, skeletal muscle and nervous tissues do not possess the very high repair capacities that occur in more mitotically active tissues. Accordingly, oxidative stress may accumulate with age in these tissues and contribute to the development of sarcopenia; however, this possibility has not been thoroughly investigated. Although long-term intervention studies which test the hypothesis that oxidative stress contributes to sarcopenia have not been conducted, recent findings using the caloric restriction (CR) paradigm for studying retarded aging processes indirectly support this concept. Several research directions appear important to pursue, including the measurement of free radical production in skeletal muscle at diverse ages, antioxidant supplementation as an intervention for retarding the development of sarcopenia, the use of genetically manipulated animal models, and determining the influence of CR on oxidative stress in specific skeletal muscles and individual fibers and neurons therein. PMID- 7493212 TI - Muscle performance and structure in the elderly as studied cross-sectionally and longitudinally. AB - Longitudinal and cross-sectional studies of muscle performance and structure in the elderly are summarized in this article. Muscle strength and muscle mass decrease with age, mainly due to the loss of muscle fibers. In most studies of the vastus lateralis, the mean fiber area decreases only slightly with a larger reduction in the type II fiber area, with the type I fiber area being constant. However, other fiber area patterns have also been recorded, such as the preserved type II fiber area in the biceps brachii muscle, which is larger than the type I fiber area. An increase in fiber areas was recorded in 76- to 80-year-old men who maintained their physical activity level rather well. The muscle enzyme levels of the muscles studied also vary, and show adaptation with training. In particular, oxidative enzymes seem to be well maintained in old age. General conclusions concerning aging effects should not be drawn from measurements in only one muscle, and the impact of the current physical activity level of the subjects should always be taken into account. PMID- 7493213 TI - Measurement of skeletal muscle: laboratory and epidemiological methods. AB - Despite skeletal muscle's central role in the pathogenesis of sarcopenia, measurement methods remain underinvestigated and inadequately validated. Our review indicates that skeletal muscle (SM) measurement methods quantify different components and properties of muscle, ranging from the atomic to whole-body levels of body composition. Laboratory methods tend to measure whole body SM (e.g., total muscle protein, muscle cell mass, and adipose tissue-free SM components) while epidemiological methods tend to measure regional muscle (e.g., anatomic SM of an extremity). Advances in computerized axial tomography and magnetic resonance imaging methods now allow accurate estimates of whole body and regional SM and promise to finally permit comprehensive in vivo studies of SM biology and methodology. These imaging methods may help to resolve many of the confusing issues that surround the investigation of this major body composition component. PMID- 7493214 TI - Muscle biopsy as a tool in the study of aging. AB - The needle biopsy procedure provides a minimally invasive means of obtaining small samples of skeletal muscle from human volunteers. Such samples can be used to examine a variety of structural and functional characteristics of muscle, including fiber type and size, capillarization, enzymatic capacities, energy substrate or protein/mRNA concentrations, metabolic responses, and contractile properties. In conjunction with other methods, biopsy sampling can also be used to estimate total muscle mass and fiber number, and to determine rates of protein synthesis and degradation. Optimal handling and storage conditions vary widely, but in general, most of the above measurements can be made using frozen tissue, so that samples can be stored almost indefinitely. The procedure is also safe and generally well-tolerated, making it possible to perform longitudinal studies of the same person. The biopsy technique is therefore well suited for examining the underlying physiological mechanisms responsible for muscle wasting in the elderly, as well as for assessing the effects of nutritional, hormonal, and/or lifestyle (e.g., exercise) interventions intended to combat this problem. Although sample size limitations have been largely overcome by the development of microtechniques, more information is needed on how to minimize the variability introduced by studying only a small fraction of the whole muscle. Studies are also required to determine whether it is sufficient to biopsy only one muscle (and if so, which is optimal), or whether there are differential effects of aging in various muscle groups that would preclude extrapolating from one muscle to all muscles in the body.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7493215 TI - From muscle properties to human performance, using magnetic resonance. AB - Our goal is to show how muscle properties can be used to understand the exercise performance limitations of the elderly. We show that magnetic resonance (MR) imaging and spectroscopy are useful for noninvasively characterizing the structural and energetic properties of muscle in vivo. Determination of muscle volume and cross-sectional area is easily and rapidly accomplished by applying quantitative morphometric methods to MR images. New MR spectroscopic techniques provide a noninvasive "biopsy" of the oxidative, glycolytic, and contractile capacities of muscle fibers. We show how the structural and energetic properties measured by MR can be used to define the functional capacity of muscle and the contribution of this capacity to the performance of the whole body (e.g., VO2max). Finally, we relate these laboratory measures of muscle properties and performance to activities meaningful to the functioning of the elderly in everyday life, such as sustained walking and stair climbing. PMID- 7493216 TI - Age, gender, and muscular strength. AB - Muscular strength can be measured by cable tensiometry, non-motorized dynamometry (e.g., handgrip dynamometer), motorized dynamometry, or with free weights or exercise machines. Advantages and disadvantages of each are discussed. Cross sectional studies indicate that isometric and concentric strength levels peak between the second and third decade, remain unchanged until the fourth or fifth decade, and start to decline from about the fifth decade at a rate of 12% to 15% per decade until the eighth decade in men. Greater strength losses in both men and women have been suggested from the few longitudinal studies available on this topic. However, concentric strength levels in women tend to peak sooner, start to decline earlier, and decrease at a slightly slower rate than men. Age- and inactivity-induced sarcopenia explains some but not all of the losses in strength with age. There is a need to determine the separate roles of disease, inactivity, and normal aging on these losses. New findings from the Baltimore Longitudinal Study of Aging suggest a preservation of eccentric strength levels with age in women. PMID- 7493217 TI - Techniques of assessing muscle mass and function (sarcopenia) for epidemiological studies of the elderly. AB - For epidemiological studies, reliable and valid measurements or indices of muscle mass are needed to screen for those elderly persons at risk for sarcopenia, and to determine the prevalence of these conditions on a national level. The methods more suitable for field or epidemiological settings are anthropometry, bioelectrical impedance, and grip strength. These field methods, while reliable, have limited demonstrated validity in their application to the elderly and are restricted by the limited functional capabilities of the elderly, which decrease with age. Epidemiological studies of the elderly need to include sufficient numbers of persons of different ethnic backgrounds, and at the older ages. For an epidemiological study of the elderly today, one also needs to select the categories of elderly persons to be included in such studies: healthy, sick, very old, handicapped, level of functional status, etc., because relationships among variables can have different statistical and biological associations. Future work should be directed at improving the design and methodology of epidemiological and serial studies so as to maximize the numbers and categories of elderly persons who can be studied. PMID- 7493218 TI - What is sarcopenia? AB - Advancing adult age is associated with profound changes in body composition, the principal component of which is a decrease in skeletal muscle mass. This age related loss in skeletal muscle has been referred to as sarcopenia. Age-related reduction in muscle is a direct cause of the age-related decrease in muscle strength. Muscle mass (not function) appears to be the major determinant of the age- and sex-related differences in strength. This relationship is independent of muscle location (upper vs lower extremities) and function (extension vs flexion). Reduced muscle strength in the elderly is a major cause for their increased prevalence of disability. With advancing age and extremely low activity levels seen in the very old, muscle strength is a critical component of walking ability. The high prevalence of falls among the institutionalized elderly may be a consequence of their lower muscle strength. Daily energy expenditure declines progressively throughout adult life. In sedentary individuals, the main determinant of energy expenditure is fat-free mass, which declines by about 15% between the third and eighth decade of life, contributing to a lower basal metabolic rate in the elderly. Data indicate that preservation of muscle mass and prevention of sarcopenia can help prevent the decrease in metabolic rate. In addition to its role in energy metabolism, skeletal muscle and its age-related decline may contribute to such age-associated changes as reduction in bone density, insulin sensitivity, and aerobic capacity.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7493219 TI - The relationship of strength to function in the older adult. AB - Reduced lower extremity strength has been associated with reduction in gait speed, balance, stair-climbing ability, and getting up from a seated position. The relationship of lower extremity strength and the ability to accomplish selected functional activities was examined in 16 healthy but frail older adults ranging in age from 75 to 88 years (mean = 80.9 years). The following measures were obtained for each subject: preferred gait speed under laboratory and free walking conditions, 5 timed chair stand-ups, and time to complete an obstacle course. Strength measures of the hip extensors, hip abductors, knee extensors, planter flexors, and dorsiflexor muscle groups were obtained using a hand-held dynamometer. The relationship between the time to complete the functional activities and each of the strength variables was determined using Pearson product moment correlations. In addition, performance was examined in relation to various combinations of strength measures (e.g., hip and knee extension). Weak, nonsignificant hip, knee and ankle strength/functional activity relationships were found for all of the variables examined. When hip extension, knee extension, and ankle plantar flexion strength values were combined and normalized to body weight, a significant strength-to-functional activity relationship was found for 14" chair stand-ups (r = .636, p < .01). When values for quadriceps strength and gait speed for 35 adults ranging in age from 60-72 years were compared to those for 75-88 year olds, marked differences emerged. A more significant relationship between knee extension force and gait speed was observed for the younger adults (r = .528 vs r = .353).(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7493220 TI - Muscle function and mobility biomechanics in the elderly: an overview of some recent research. AB - Many old adults have difficulty in performing activities of daily living, in maintenance of postural balance, and in recovering from impending falls. It is not yet fully clear to what extent these difficulties arise from age, or disease related declines in muscle function. The strength requirements for the performance of many common physical tasks are not often large. When the time available to make an appropriate response is short, maximum joint torque strengths may not be as important a consideration as abilities to develop joint torques rapidly. Even old adults who are fit and healthy, compared to young adults, have substantially diminished abilities to do this. Recent findings suggest that the source of this decline, at least sometimes, lies in muscle physiology more so than in central processing delays. A considerable amount of research to explore the issues relevant to the relations among muscle function and mobility is currently underway, but much remains to be learned. PMID- 7493221 TI - Strength is a major factor in balance, gait, and the occurrence of falls. AB - We studied the effects of lower extremity strength as well as gait and balance on the occurrence of falls in nursing home residents. Nursing home residents with a history of falls had less than half of the knee and ankle strength of nonfalling subjects residing in the same home. The differences were more prominent at the ankle than the knee, and were most pronounced in the ankle dorsiflexors, where they were one-tenth that of controls. Also of note was the fact that this same group of fallers had slowed gait velocity (58% of control) as well as an impaired response to postural perturbation as determined on the Postural Stress Test (55% of control). In a recently completed study we measured strength as balance (EquiTest balance platform) of community-dwelling subjects. The occurrence of loss of balance during the sensory organization test was correlated with diminished lower extremity (Pearson R = -.36, p = .001) as well as ankle dorsi and plantar flexion moments (Pearson R = -.37, p = .001). Using a logistic regression model, we demonstrated an independent effect of strength on the odds ratio of an SOT-LOB; for each newtonmeter per kg increase in strength there was a 20% decrease (95% CI, .74-87) in the odds ratio. The data from both nursing home and community-dwelling subjects indicate a strong relationship of lower extremity strength to balance and gait. The nursing home studies demonstrated an association between these functions and the occurrence of falls.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7493222 TI - Loss of skeletal muscle mass with aging: effect on glucose tolerance. AB - It is generally believed that aging results in glucose intolerance. Since skeletal muscle is the major site of glucose uptake following an oral glucose tolerance test (OGTT), it is reasonable to hypothesize that loss of muscle mass with aging causes glucose intolerance. Evidence against this concept comes from extensive data demonstrating the usefulness of the recommended procedures for performing an OGTT. This involves giving 75 g of glucose to all subjects regardless of body weight, and the criteria for categorizing glucose intolerance are the same for all individuals. That this works well, with no evidence that larger people have better glucose tolerance than smaller people, suggests that muscle mass does not usually play a role. Recent studies on humans and rats indicate that aging per se does not result in glucose intolerance. In most of those older people with decreased glucose tolerance, this problem appears to be due to accumulation of abdominal fat with development of insulin resistance. We conclude that the loss of muscle mass with aging does not usually result in glucose intolerance. PMID- 7493223 TI - Sarcopenia in aging humans: the impact of menopause and disease. AB - We examine the association of the menopause transition, congestive heart failure, and Parkinson's disease on body composition and energy expenditure. We present evidence suggesting that the normal menopausal transition is associated with accelerated loss of fat-free mass, a decline in resting metabolic rate, and increased central body fatness. Second, we show that the cardiac cachexia associated with heart failure is partially due to an elevated level of energy expenditure. Despite having a lower quantity of fat-free mass, congestive heart failure patients have a higher resting metabolic rate (approximately 283 kcal/d) for their metabolic size than healthy elderly. The elevated level of resting energy expenditure probably contributes to their unexplained weight loss. Parkinson's patients experience muscular rigidity and tremor which could contribute to inappropriately high levels of energy expenditure and difficulty in maintaining body weight and composition. We examined resting metabolic rate and body composition in eight Parkinson's patients and 34 healthy age-matched controls. Parkinson's patients showed lower levels of fat-free mass (approximately 6 kg), but similar resting metabolic rates (1601 +/- 250 kcal/d) versus healthy controls (1671 +/- 212 kcal/d), suggesting a hypermetabolic state. A re-examination of daily energy needs and the metabolic factors contributing to periods of energy imbalance during the menopausal transition and in several disease states may be a prerequisite to offsetting accelerated sarcopenia. PMID- 7493224 TI - Functional consequences of sarcopenia: effects on thermoregulation. AB - The loss of skeletal muscle mass with aging (sarcopenia), and related changes in body size and composition, may impact body temperature and thermoregulation in both hot and cold environmental conditions. Sarcopenia alters the thermal properties of the body as a passive system because of differences in water content, and thus specific heat, of muscle and adipose tissue. With respect to active thermoregulation in warm environments, differences in fat-free weight (FFW) can explain more than 80% of the variance in absolute blood volume (BV) among individuals (Allen et al., 1956) and BV, in turn, profoundly influences the cardiovascular responses to exercise and heat stress. For example, a lower BV for a given body weight may explain more than half of the variability in maximal oxygen uptake (VO2max). Thus, as VO2max declines, any absolute task represents a higher relative VO2max (% VO2max) and proportionately greater cardiovascular strain. Because BV is an important determinant of left-ventricular filling pressure, and because older individuals rely more on the Frank-Starling mechanism to increase cardiac output (Q), a lower BV may also be associated with an inability to increase Q appropriately. These effects are particularly important under conditions of heat stress, where a larger increase in Q is necessary to perfuse both skin and active muscle vascular beds. With exposure to cold, age related changes in body composition affect the insulation provided by the peripheral tissues, especially in the limbs. This results in an increased reliance on peripheral vasoconstriction to minimize heat loss, yet this vasoconstriction is attenuated in older individuals.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7493225 TI - Relationship of age-related decreases in muscle mass and strength to skeletal status. AB - In this presentation, age-related bone loss is seen to be the consequence of a small yet persistent insufficiency in bone remodeling, where small deficits in bone mass remain upon completion of each remodeling cycle. Although loss of bone and of muscle strength develop together over time, remodeling imbalance appears to emerge much earlier than does loss of muscle mass and strength. Thus, age related bone loss cannot be ascribed entirely to the loss of muscle. Although muscular action does influence bone mass, its most impressive effects occur between complete immobilization and ambulatory, but sedentary life. Regardless of the minor role sarcopenia may play in bone loss, muscle weakness exerts a powerful influence on hip fracture incidence because of its effect on the risk of falls. PMID- 7493226 TI - Motor units: remodeling in aged animals. AB - The effects of aging on motor units of different types and the spatial arrangement of motor units are discussed. This article is, however, not a comprehensive survey of the literature, but rather a summary of our own results with the addition of relevant reports. The glycogen depletion technique was used to identify the muscle fibers in a motor unit (motor unit fibers). The motor unit fibers were subsequently characterized according to immunocytochemical, enzyme histochemical and morphometrical properties, and the spatial arrangement was determined with the aid of a computer-assisted model. An age-related motor unit transformation was observed in both fast- and slow-twitch rat skeletal muscles as was a rearrangement of motor unit fibers. The spatial redistribution of motor unit fibers, higher innervation ratios, and larger motor unit territories in old age indicate an age-related denervation-reinnervation process due to loss of alpha-motoneurones. PMID- 7493227 TI - Factors influencing the repair and adaptation of muscles in aged individuals: satellite cells and innervation. AB - From the standpoint of structure, the loss of muscle mass could be attributed to the loss of muscle fibers, the reduction of volume of persisting muscle fibers, or both. This review will concentrate upon factors that could contribute to the presence of thin muscle fibers in old muscles. One mechanism that could account for the presence of a population of thin muscle fibers is the loss of innervation due to the death or the remodeling of motor units in aging muscles. If the denervated muscle fibers fail to become reinnervated, or if they are unable to respond to reinnervation by new nerve terminals, they would undergo a progressive atrophy. If muscle fibers are damaged through exercise, direct trauma or other causes, they would degenerate and regenerate. Early regenerating muscle fibers are thinner than normal. If the regenerating muscle fibers are non-innervated, regeneration will not be complete; instead the regenerating muscle fibers will atrophy. All of the above mechanisms could contribute to an overall reduction in muscle mass. This review ends with an enumeration of outstanding questions concerning muscle atrophy and its reversal in old individuals. PMID- 7493228 TI - Physiology of cumulative defect curves; consequences in glaucoma perimetry. AB - Purely diffuse field loss is a controversial sign of early glaucoma. Cumulative defect curves have been proposed to distinguish between diffuse and localized visual field loss. In such curves, pointwise deviations from the normal reference field are ranked. The deviations are then compared with normative limits for each rank. The ranking leads to a complete loss of spatial information and ignoration of pointwise physiological threshold variability. We believed that this might impair the curves' intended ability to distinguish diffuse and localized types of defects. The lower 5% normative limit was estimated for each rank in the cumulative defect curve using Humphrey 30-2 tests of 88 subjects from the Statpac normal database. For each rank we identified those two test points (among the total of 6512 (74 test points in each of 88 subjects)) from which the 5% limit was calculated. The average eccentricity, eccrank, of those two points was then determined at each rank. For lower to intermediate ranks eccrank was spread out from center to periphery. When instead the 20 highest ranks (most depressed points) were considered eccrank was much concentrated peripherally. Our normative limits were subsequently used to evaluate field results in 101 patients with glaucoma. In this glaucoma evaluation group, the largest deviations were often encountered in central or paracentral scotomas, in sharp contrast to the more peripheral position in the normative group. These deviations often failed to reach the cumulative defect curve limits which originated from points in the mid periphery where normal variability is large. At present, cumulative defect curves are not optimized to distinguish diffuse and localized field loss. PMID- 7493229 TI - Peripheral contrast sensitivity for dynamic sinusoidal gratings in early glaucoma. AB - Automated contrast thresholds were established for dynamic, 2 reversals per sec phase-shifted, sinusoidal gratings in a 5 degrees circular test field centred 7.5 degrees from the fixation point in six positions in the visual field in 16 patients (age 56-76 years) with clinically established (optic disc or nerve fibre layer damage) early glaucoma, without visual field defects. For reference, contrast thresholds were also obtained in 16 sex-matched and age-matched controls. For all three spatial frequencies, thresholds were raised in the glaucoma group (0.5 c/deg: p = 0.0034-0.041, 2 positions n.s.; 1.0 c/deg: p = 0.0093-0.049, 1 position n.s.; 2.0 c/deg: p = 0.0011-0.041, 1 position n.s.). Deficiencies did not correlate to glaucoma type (simplex vs capsular), intraocular pressure level at examination, visual acuity, or local therapy (beta blockers vs miotics). In the glaucoma group, there was a general loss of contrast sensitivity in all six positions tested in the visual field, reflected in roughly similar threshold elevations. Losses appeared slightly greater in the lower hemi field. Definite contrast threshold elevations from the early glaucomatous process were established in this group of glaucoma patients without visual field defects. However, the method was considered to be of less value in glaucoma detection for individual subjects, due to the large overlap in threshold values between glaucoma patients and controls. PMID- 7493230 TI - Primary argon laser trabeculoplasty vs pilocarpine. III. Long-term effects on visual fields. AB - In a prospective randomized study on primary argon laser trabeculoplasty vs pilocarpine in 82 newly detected open-angle glaucoma patients, visual field changes were analysed for absolute changes over 2 years on automated threshold perimetry and manual Goldmann perimetry. Regression analysis was also performed using the threshold perimetry examinations taken every second month. Analyses based on automatic perimetry were corrected for eventual bias due to cataract and induced miosis. There was significantly less decay of visual field in the group treated with laser. This was even more pronounced for eyes with capsular glaucoma, especially in the early stage. For the smaller group with simple glaucoma no significant advantage of laser could be shown. Successful intraocular pressure reduction significantly improved the visual field outcome. Laser trabeculoplasty is therefore recommended as primary therapy in capsular glaucoma. PMID- 7493231 TI - Primary argon laser trabeculoplasty vs pilocarpine. IV. Long-term effects on optic nerve head. AB - In a prospective, randomized study on primary argon laser trabeculoplasty vs piloparpine in 82 newly detected open-angle glaucomas the progress of optic nerve head damage is presented. In 70 patients reliable stereo photographs could be analysed. In the laser treated group a progression of glaucoma damage after 24 months was seen in 6/37 eyes (16%) and in the pilocarpine-treated group 16/33 (49%) (p = 0.0048). There was a statistically significant favourable outcome if intraocular pressure was regulated in terms of peak and range of pressure, but not average pressure. This was even more pronounced in capsular glaucoma, but not significant for simple glaucoma. Pilocarpine increased the risk for further damage to the optic nerve head 6.4 times compared to laser trabeculoplasty. Thus, laser is recommended as the primary therapy in high tension glaucoma, especially in capsular glaucoma. PMID- 7493232 TI - Prevalence of glaucoma and distribution of intraocular pressure in a population. The Casteldaccia Eye Study. AB - IOP was measured in 1062 middle-aged and elderly subjects of a small Sicilian town, enrolled in a population based survey. The mean IOP was 15.1 +/- 3.7 mmHg without interocular or sex differences. A small but significant age-dependent increase of IOP was found. Circadian and seasonal influences were recorded: IOP was higher in the morning and in winter. The prevalence of chronic open-angle glaucoma was 1.2%, but it grew to 3.6 if only subjects aged 70 years or more were considered. IOP of 24 mmHg or more was found in 2.7%, while 4.3% of subjects showed IOP of 21 mmHg or more. This survey shows that mean IOP of the Mediterranean population studied is similar to the IOP found in other epidemiologic investigations; the prevalence of glaucoma, however, seems to be rather high. PMID- 7493234 TI - The short-time effect of latanoprost on the intraocular pressure in normal pressure glaucoma. AB - Latanoprost, a prostaglandin analogue, was given topically to 20 patients with normal pressure glaucoma in a double masked randomized study. Either latanoprost 0.006% or placebo (vehicle) was administered twice a day for 14 days. Latanoprost caused a statistically significant (p < 0.001) reduction in intraocular pressure from a diurnal baseline level of 16.8 to 14.3 mmHg, as measured on day 14. Latanoprost was well tolerated. PMID- 7493233 TI - The amount of lens exfoliation and chamber-angle pigmentation in exfoliation syndrome with or without glaucoma. AB - Correlations of the amount of chamber-angle pigmentation and exfoliation material with IOP and with image analysis of the optic discs are reported in the paired eyes of 63 persons with unilateral exfoliation syndrome, and 35 patients with unilateral capsular glaucoma. The effect on IOP of phenylephrine-induced pigmentary dispersion is examined. Chamber-angle pigmentation was more pronounced in capsular glaucoma than in the nonglaucomatous exfoliation eyes (p < 0.01), although the amount of lens exfoliation was the same. In nonglaucomatous exfoliative eyes IOP correlated significantly with the amount of chamber-angle pigmentation (p < 0.01) and exfoliation material (p < 0.05). The nonglaucomatous exfoliative eyes with grade > or = 2 trabecular pigmentation and those with reduction of IOP on phenylephrine provocation had smaller rim areas and rim/disc area ratios than the fellow eyes (p < 0.05). Rather than the amount of exfoliation, the main risk factor for glaucoma seems to be the degree of chamber angle pigmentation. PMID- 7493235 TI - Anatomic features of the optic disc and risk of progression in ocular hypertension. AB - We evaluated characteristics of the optic disc in ocular hypertensive patients which might portend future glaucomatous damage. We included in this study 12 patients with an intraocular pressure > 21 mmHg who on follow-up showed signs of optic nerve head damage. Each of these patients were matched to an ocular hypertensive patient who remained stable for > 5 years. This study found that neither optic disc, neural rim or peripapillary halo areas, vein or artery diameter over non-atrophied or atrophied retinal areas, differed significantly between groups (p > 0.05, Wilcoxon signed rank test). However, peripapillary atrophy was greater in patients who showed signs of glaucomatous disc damage (1.757 +/- 0.36 mm2, p = 0.02, Wilcoxon signed rank test) compared to stable patients (1.064 +/- 0.79 mm2), although this finding was not significant after the Bonferroni correction. Patients with < 0.6 mm2 area of peripapillary atrophy (n = 6), however, did not suffer damage. This study suggests that a relationship between the extent of peripapillary atrophy and the chance of developing optic disc damage in patients with ocular hypertension deserves further study. PMID- 7493237 TI - Effects of antiglaucoma drugs on ocular surface. AB - Topical medications have a very important role in chronic glaucoma treatment. Long-term use of these medications can cause significant changes on ocular surfaces. In this study, the ocular surfaces of 20 control subjects (group I), 20 primary open-angle glaucoma patients (group II) treated (mean 21.20 +/- 1.32 months) with 0.50% timolol maleate, and 20 primary open-angle glaucoma patients (group III) treated (mean 21.70 +/- 1.34 months) with 0.50% timolol maleate + 1% dipivefrin hydrochloride were evaluated. Studied parameters included Schirmer's test, tear break-up time, conjunctiva impression cytology and goblet cell density. These results suggest that long-term applications of topical anti glaucoma medications damage the ocular surface. PMID- 7493236 TI - The morphology of conjunctiva after long-term topical anti-glaucoma treatment. A quantitative analysis. AB - On biopsy material, differences in the degree of conjunctival inflammation and fibrosis between topically treated glaucoma patients and age matched controls, were examined histologically. Eighteen patients with primary glaucoma underwent goniotrephinations, because maximal medical therapy had failed. The patients had received at least two types of topical anti-glaucoma drugs for at least 12 months (mean 46 months). The control group consisted of 18 age-matched control patients without glaucoma, who had received no topical therapy. These patients underwent cataract surgery or squint surgery. Biopsies were taken from the infero-temporal bulbar quadrant with a biopsy forceps. The specimens were fixed while stabilized on a rubber support to exclude any major shrinkage. Specimens were analyzed by light microscopy for the content of inflammatory cells (plasma cells, lymphocytes, polymorphs, macrophages and mast cells), goblet cells and fibroblasts. No significant difference in the histologic parameters between the two groups could be demonstrated. The study suggests that topical treatment for periods up to 4 years with anti-glaucoma drugs does not induce morphological signs of inflammation and fibrosis of the conjunctiva. PMID- 7493238 TI - IgG, albumin and total IgG index in the aqueous humour of eyes with pseudoexfoliation syndrome. AB - IgG and albumin in the aqueous humour and serum were studied using kinetic nephelometry, carried out on 34 eyes with pseudoexfoliation and 31 normal eyes (control group). The results show a rupture in the hemato-aqueous barrier for IgG and albumin in the group of eyes with pseudoexfoliation and glaucoma, and only for albumin in the group of eyes with pseudoexfoliation without glaucoma. The calculation of the total IgG index rules out the existence of a primary or associated immunological mechanism in eyes with this syndrome. PMID- 7493240 TI - Early hypotony after trabeculectomy. AB - The occurrence of early hypotony after trabeculectomy was analysed retrospectively in 60 glaucoma patients. Fifty-two per cent of the eyes had an intraocular pressure < or = 10 mmHg on the first postoperative day. In about one third of the eyes, the intraocular pressure was < or = 5 mmHg at the first postoperative visit and in more than 70% of these eyes the hypotony was almost unchanged one week after operation. The hypotony one week after operation was not correlated to the age of the patients and the intraocular pressure at operation, nor to the glaucoma type, and showed no significant statistical dependence on the depth of anterior chamber and hyphema. The final untreated intraocular pressure and progression of the postoperative cataract were studied in four groups of patients, formed on the basis of the intraocular pressure level one week after operation: patients with marked hypotony, slight hypotony, normal pressure or with hypertension. The final untreated intraocular pressure in the eyes with early marked hypotony was not significantly different from the final untreated intraocular pressures in the slightly hypotonic and normotonic eyes. However, the untreated intraocular pressures in these three eye groups were significantly different from the untreated intraocular pressure in the eye group with hypertension. Fifty-two per cent of the eyes suffered from cataract progression during a mean follow-up period of 24 months. Postoperative cataract progression in the markedly hypotonic eyes was not significantly different from the cataract progression in the other groups of eyes. PMID- 7493242 TI - Reproducibility of the Glaucoma-scope in cadaver eyes. AB - We evaluated the intra- and inter-observer variability of depth measurements of the optic disc in cadaver eyes using the Glaucoma-scope. Intra-observer variation of 10 separately captured images became significantly greater with increasing depth of the optic cup (p < 0.001). The standard deviation of depth measurements were 9.06 mu over the neural rim, 25.00 mu along the cup wall, and 40.94 mu at the bottom of the optic cup. However, intra-observer variation of a single captured image did not increase with greater optic cup depth. Intra-observer variation, but not inter-observer variation, generally was greater when measuring the superior than inferior optic disc (p < 0.05). This study suggests that when capturing separate optic disc images with the Glaucoma-scope, increased variation in depth measurements can be expected with greater optic cup depth and in the superior portion of the optic disc. PMID- 7493239 TI - Traumatic glaucoma. AB - In the management of an ocular injury, traumatic glaucoma is often overlooked. Prospectively reviewing 100 consecutive patients of traumatic glaucoma we found a very high correlation of postconcussional glaucoma with traumatic cataracts, angle recession of more than 180 degrees, significant injuries to the iris and a displacement of the lens. Two of these four clinical features were present in all cases. Penetrating injuries were followed by a secondary glaucoma if an adherent leucoma and/or evidence of lenticular damage or displacement were seen. These clinical features all point to a damage to the lens-iris diaphragm. The trabecular meshwork being directly related to these structures would also be involved in such an injury, leading to significant impairment of function, and the eye would therefore be more prone to developing glaucoma. Our study has highlighted certain clinical findings which could act as 'markers' for the early diagnosis of traumatic glaucoma. PMID- 7493241 TI - Medical and surgical aspects of congenital glaucoma. AB - During the years 1977-1986, 338 patients at the University Eye Clinic were diagnosed as having congenital glaucoma. This paper presents our experience of the management of 88 of these patients (161 eyes) who had a follow-up of at least 3 years. Medical therapy alone reduced the intraocular pressure to less than 21 mmHg in 17 eyes (11.8%) in the short-term and in 14 eyes (9.7%) in the long-term. Surgical intervention in 127 eyes (goniotomy: 9 eyes; trabeculotomy: 3 eyes; peripheral iridencleisis: 4 eyes; Elliott trephine; 23 eyes; trabeculectomy: 88 eyes), resulted in an immediate normalization of intraocular pressure in 98 eyes (77%, reducing to 66% at the final examination). Trabeculectomy normalized the intraocular pressure in 84% of eyes in the short-term and 76% in the long-term. Trabeculectomy is recommended as the surgical management of choice in congenital glaucoma in societies where presentation is late, or where individual surgeons may not be conversant with goniotomy. PMID- 7493244 TI - Indications and efficacy of combined trabeculectomy and extracapsular cataract extraction with intraocular lens implantation in cataract patients with coexisting open angle glaucoma. AB - In an observational study 19 eyes with cataract and coexisting primary open angle glaucoma were scheduled to undergo combined procedure with ECCE-IOL and trabeculectomy. The aim of the present study was to investigate the results and the efficacy of combined surgery, when we extended the indication for surgery. Thus the surgical decision making was not based solely upon IOP readings, but also involved social/psychological factors. Median decrease in IOP (IOPpost IOPpre) was significant through a 12-month observation period, p < 0.001. The number of medications dropped from 1.74 to 0.21, p < 0.01. Complication rate during operation and in the postoperative follow-up period was low. We conclude that it seems justified to extend the indication for combined surgery in cataract patients with coexisting open angle glaucoma in case of uncontrolled IOP, poor compliance, inability of sufficient medical care, or unacceptable medication. We recommend careful monitoring of the early postoperative IOP, even if hypotensive agents are given prophylactically. PMID- 7493243 TI - The effect of flat anterior chamber on the success of trabeculectomy. AB - The authors reviewed 70 eyes of 59 patients who underwent trabeculectomy. We divided the eyes into the flat anterior chamber group (23 eyes) and formed anterior chamber group (47 eyes, control group). The influence of clinical variables on the development of flat anterior chamber, and the effect of flat anterior chamber on the success of trabeculectomy were evaluated. The two groups were similar in terms of many variables, but the preoperative intraocular pressure (27.1 +/- 15.4 vs. 18.1 +/- 8.7 mmHg), incidence of cataract progression (9/18 vs. 10/44) and choroidal detachment (5/23 vs. 2/47) were higher in the flat anterior chamber group (p < 0.05). The overall success rates for the follow-up period of 30 months were similar between the two groups (p > 0.05). However, the short-term (2 to 8 months) postoperative success rates were lower in the flat anterior chamber group (p < 0.05). The development of flat anterior chamber following trabeculectomy can be reduced by lowering the preoperative intraocular pressure, and flat anterior chamber may be harmful to the postoperative course of trabeculectomy. PMID- 7493245 TI - Aqueous shunts: single-plate Molteno vs ACTSEB. AB - A retrospective study was done to compare the results of the single-plate Molteno implant surgery with those of the anterior chamber tube shunt to an encircling band procedure in eyes with refractory glaucoma. Both Molteno implant and anterior chamber tube shunt to an encircling band are made of silicone. There were 19 eyes in the Molteno group, 17 eyes in the anterior chamber tube shunt to an encircling band group, with at least 6 months of follow-up. Preoperative intraocular pressure in the Molteno group and anterior chamber tube shunt to an encircling band group was 47.5 +/- 11.4 and 46.5 +/- 10.3 mmHg, respectively. Intraocular pressure at 6 months postoperatively was 20.4 +/- 15.1 and 15.5 +/- 10.4 mmHg, respectively. The difference in pressure reduction between the two groups was not significant statistically. Serious complication rate for 6 months was 11% and 47%, respectively. PMID- 7493246 TI - Intraocular pressure elevation caused by massive and prolonged hemorrhage after trabeculotomy ab externo. AB - A case with postoperative elevated intraocular pressure levels caused by massive and prolonged hemorrhage after trabeculotomy ab externo is presented. In this case, intraocular pressure was controlled by removal of the hemorrhage, but the transient elevation resulted in progression of visual field defect. Thus, resorption of hyphema and intraocular pressure levels should be monitored closely after trabeculotomy, and surgical removal of the hemorrhage appears to be an effective treatment for this rare complication. PMID- 7493247 TI - Argon laser photocoagulation of cyclodialysis clefts after cataract surgery. AB - Three patients with cyclodialysis clefts, hypotony and hypotonic retinopathy subsequent to cataract surgery were treated with argon laser photocoagulation. The hypotony was reversed in each patient and their visual acuity was normalized. Laser photocoagulation is a noninvasive treatment that can be repeated easily and safely. The complications of the treatment are minor. A hypertensive episode commonly occurs in the early postoperative period. PMID- 7493248 TI - Inadvertent instillation of Minims eye drops. PMID- 7493249 TI - Thermal aspects of vehicle comfort. AB - The combined thermal effects of convection, radiation and conduction in a vehicle compartment need special measuring equipment accounting for spatial and temporal variations in the driver space. The most sophisticated equipment measures local heat fluxes at defined spots or areas of a man-shaped manikin. Manikin segment heat fluxes have been measured in a variety of vehicle climatic conditions (heat, cold, solar radiation etc.) and compared with thermal sensation votes and physiological responses of subjects exposed to the same conditions. High correlation was found for segment fluxes and mean thermal vote (MTV) of subjects for the same body segments. By calibrating the manikin under homogenous, wind still conditions, heat fluxes could be converted (and normalised) to an equivalent homogenous temperature (EHT). Regression of MTV-values on EHT-values was used as basis for the derivation of a comfort profile, specifying acceptable temperature ranges for 19 different body segments. The method has been used for assessment of the thermal climate in trucks and crane cabins in winter and summer conditions. The possibility for spatial resolution of thermal influences (e.g. by solar radiation or convection currents) appeared to be very useful in the analysis of system performance. Ventilation of driver's seats is a technical solution to reducing insulation of thigh, seat and back areas of the body. Constructions, however, may vary in efficiency. In one system seat ventilation allowed for almost 2 degrees C higher ambient conditions for unchanged general thermal sensation, in addition to the pronounced local effect. In a recent study the effects of various technical measures related to cabin design and HVAC systems have been investigated.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7493250 TI - Physical fitness and cardiopulmonary baroreflex to passive head-up tilt. AB - The relationship between physical fitness and cardiopulmonary (CP) baroreflex induced by the decrease in venous return was investigated. Maximal oxygen uptake/lean body mass (VO2max/LBM) was used as a measure of physical fitness. CP baroreflex was examined in 8 subjects [22.0 +/- 2.0 (SD) yr] ranging from low (47.3 ml.kg-1.min-1) to highly (72.6 ml.kg-1.min-1) fit subjects who were exposed to the orthostatic load using passive head-up tilt. Tilting from 0 degrees to 70 degrees in 10 degrees increments was applied. Each tilting was preceded by measurements at the basal position of 0 degrees. Heart rate, systolic and diastolic pressure, stroke volume, and forearm blood flow were measured before and during tilting; Cardiac output, mean arterial pressure, and forearm vascular resistance (FVR) were calculated. FVR was increased with the raising in tilt angle in each subject, but the onset of increase in FVR tended to be earlier in proportion to the higher level of physical fitness. There is a negative and significant correlation between the onset of increase in FVR and VO2max/LBM. The physically fit subject exhibited a more rapid onset of an increase in FVR to orthostatic stress than the unfit subject. Thus the higher the physical fitness, the lower the threshold in CP baroreflex response. PMID- 7493251 TI - Adjustments of cardiac function during prolonged exercise relative to lactate threshold. AB - Alterations in cardiac function during prolonged submaximal exercise relative to lactate threshold (LT) were evaluated on 7 normal healthy males aged 30.3 +/- 5.7 years. Systolic time intervals were analyzed through simultaneous recordings of electrocardiogram, phonocardiogram, and impedance cardiogram at a paper speed of 50 mm/s. Determination of stroke volume (SV) was based upon the method described by Kubicek et al. ANOVA followed by the Scheffe post-hoc comparison revealed that SV and myocardial contractility indices (MSER and PEP/LVET) remained relatively unchanged throughout 40-min cycling exercise, although the changes in heart rate and oxygen uptake were statistically significant. In addition, systolic blood pressure remained almost unchanged during the exercise. These results may be interpreted as evidence of the "contractility reserve", i.e., the ability of maintaining heart muscle contractility during prolonged exercise at LT intensity. PMID- 7493252 TI - Total body fat estimations by near-infrared interactance, A-mode ultrasound, and underwater weighing. AB - We studied 94 (41 men and 53 women) healthy 18 to 65 years old Caucasians for body composition utilizing 3 methods: 1) Underwater weighing (UWW) for body density, 2) A-mode ultrasound subcutaneous fat determination, and 3) Near infrared (NIR) interactance spectral data. We then utilized the Wherry-Doolittle test selection method to predict percent of total body fat (%fat) from either ultrasound or NIR data with anthropometric measurements, optimizing for the fewest number of data points which correlated highly with UWW results. It was found that %fat could be estimated with a correlation of > 0.9 when measured at 4 sites (different between men and women) by ultrasound, age (women) and height (men). We also found a correlation of > 0.8 using 2 sites by NIR, weight, age, and height. It has been suggested that the ultrasound method would produce better estimates of both local and total body fat. PMID- 7493253 TI - Sex differences in the shapes of several parts of the young Japanese face. AB - For the purpose of studying the sex differences of the human face we collected five separate images, which consist of several parts of the face, from frontal view photographs of 48 male and 52 female college students. We traced outlines of their faces with simple lines (traced items), and made reproductions of the photographs of their eyes, mouth and nose by using a copying machine (reproduced items). The test subjects were 16 males and 8 females. They looked at parts of the face shown in each image, and categorized them individually by judging on their sex. Then, we calculated the percentages of correct judgments (percentage correct) for each image. By comparing the percentage correct between male and female we concluded that the sex of the subjects did not affect the results of their judgments. In the traced items the percentage correct for the face as a whole, which contained the outlines of the eyes, mouth, nose and the lower jaw, was 69%, but it decreased to 61% when the outline of the lower jaw was removed. Hence, the outline of the lower jaw apparently has a characteristic shape easily noticed by males. In the reproduced items the percentage correct was 65% for the eyes, 68% for the mouth and 58% for the nose. The mouth, therefore, has more distinguishing characteristics than the eyes or nose, especially with females. On the other hand, there is no correlation between the percentage correct for the eye, mouth and nose items. Hence, we concluded that the sexual specificity for the shape of the young Japanese face appears on their parts independently. PMID- 7493255 TI - Peripheral vascular anatomy, physiology, and pathophysiology. AB - The primary function of the circulatory system is to transport oxygen, nutrients, and waste products. Hemodynamics of the vessels are governed by a variety of physical properties and laws that explain blood flow through the vascular system. Blood flow is affected by pressure differences, radius of the vessel, length, and viscosity (Poiseuille's Law). Flow is directly proportional to the difference in pressure and inversely proportional to resistance. Any disturbance in blood flow in the arterial and venous system disrupts the delivery of oxygen and nutrients and the elimination of waste products. Disturbances of blood flow can be due to compression of the blood vessel (edema, hematoma, compartment syndrome), structural changes within the vessel (atherosclerosis), vasospasm (Raynaud's syndrome), vasodilatation (sepsis, distributive problems), or blood clot (thrombus or embolus). Decrease in perfusion will promote compensatory mechanisms such as vasodilatation, development of collateral vessels, and anaerobic metabolism. If compensatory mechanisms are unable to meet the oxygen demand by the tissues, ischemia develops, and eventually, tissue death occurs. PMID- 7493254 TI - A one-year follow-up study of body composition in a young woman. PMID- 7493256 TI - Hypertension: diagnosis, acute antihypertension therapy, and long-term management. AB - Hypertension remains a major modifiable risk factor for coronary heart disease, cerebrovascular disease, peripheral vascular disease, and renal disease. Although great strides have been made in increasing patient awareness, lifestyle changes, and compliance with treatment protocols, hypertensive emergencies and hypertensive urgencies remain a major concern. Unless treated promptly, irreversible target organ damage will ensue. Therefore, patients with acutely elevated blood pressure, regardless of cause, must be evaluated expeditiously, and appropriate treatment must be initiated. Nurses must be able to assess and monitor patients and their progress and recognize signs and symptoms of complications related to hypertension. Once blood pressure control has been achieved, nurses initiate patient education. However, long-term management must not only include blood pressure monitoring and patient education, but also emphasize lifestyle changes. PMID- 7493257 TI - Diabetes and vascular disease: a common association. AB - Diabetes is a chronic metabolic disorder with a characteristic hyperglycemia. This elevated blood glucose causes the frequent complications of diabetes that often involve the vascular system. Macrovascular involvement includes coronary artery disease, cerebrovascular disease, and peripheral vascular disease. Diabetic nephropathy and retinopathy are serious microvascular disturbances. There is no cure for diabetes, so early detection and intervention are necessary to limit progression of diabetes and its complications. The advanced nurse practitioner has a vital role in directing care for chronic conditions through education and a holistic approach to the patient as they are key to diabetes management. Standards of practice for diabetes have been defined by the American Diabetes Association. Balancing cost containment with maintenance of these standards is a challenge for health care. PMID- 7493258 TI - Revascularization in peripheral vascular disease: stents, atherectomies, lasers, and thrombolytics. AB - Peripheral vascular disease affects a significant number of individuals. Signs and symptoms may develop because of partial or total vessel occlusion due to plaque, dissection, or thrombus. Percutaneous transluminal angioplasty is effective as an independent intervention to open occluded vessels and also may be combined with other nonsurgical therapies such as stents, atherectomy, or laser treatments. Thrombolytics also are used to treat acute or chronic occlusions. The nurse's role in treating and monitoring the patient is key in minimizing complications during and after intervention. PMID- 7493259 TI - Perioperative management of patients with vascular disease. AB - Peripheral artery occlusive disease affects approximately 12% of the population. The evolution of vascular surgery and technologic advances has provided relatively safe and effective treatment regimens for patients with peripheral artery occlusive disease. Vascular surgeries, including carotid endarterectomy, repair of abdominal aortic aneurysm, femoral artery surgeries, thromboembolectomy, and amputation are common surgical interventions performed routinely in many operating rooms. Technologic advances have provided for the success of surgery on occluded arteries. Perioperative nursing care of the patient involves a thorough understanding of the progressive nature of the disease, surgical intervention as palliative, available technology to assist with assessment, and patient and family coping mechanisms. The role of the nurse in caring for patients with peripheral artery occlusive disease spans the health care continuum from acute care to chronic care. Perioperative, postanesthesia, and intensive care nurses focus on acute care associated with surgical intervention. Practitioners in the rehabilitative and home care arenas must find innovative methods to educate patients and families to become independent and active participants in their care. PMID- 7493260 TI - Stroke: brain attack, time to react. AB - New developments in neuroscience challenge nurses to care skillfully for patients with cerebrovascular diseases. Because stroke is the leading cause of brain damage in adults, professionals are urged to approach stroke with the same urgency as other medical emergencies. Understanding the physiologic early sequelae of brain damage is essential to appreciation of the newer therapeutic treatments under study. Cardiac and hemodynamic complications of cerebrovascular disease emphasize the important role of nurses in the assessment and protection of stroke patients. Nursing implications include public education about stroke symptoms and the urgent evaluation and management of stroke. PMID- 7493262 TI - Optimizing the flow of care for prevention and treatment of deep vein thrombosis and pulmonary embolism. AB - Critically ill patients have multiple risk factors for deep vein thrombosis and pulmonary embolism. The majority of patients with pulmonary embolism have a lower extremity deep vein thrombosis as a source of origin. Pulmonary embolism causes a high mortality rate in the hemodynamically compromised individual. Awareness of risk factors relative to the development of deep vein thrombosis and pulmonary embolism is important for the critical care nurse. Understanding the pathophysiology can help guide prophylaxis and treatment plans. The therapies, from invasive to mechanical, all carry risks and benefits, and are weighed for each patient. The advanced practice nurse, whether in the direct or indirect role, has an opportunity to impact the care of the high risk patient. Options range from teaching the nurse who is new to critical care, to teaching patients and families. Development of multidisciplinary protocols and clinical pathways are ways to impact the standard of care. Improved delivery of care methods can optimize the care rendered in an ever changing field of critical care. PMID- 7493261 TI - Case studies: what a difference a nurse makes. AB - The case studies represent three different types of critical care nursing challenges. The objective of each case study is to identify a key nursing problem and focus on nursing research or nursing behavior that made or could have made the greatest difference to the patient outcome. The first case is of a man with Buerger's disease who had unmet needs regarding pain control. The second patient had an out-of-hospital ruptured abdominal aortic aneurysm and survived a rough, 7 week hospitalization. Major complications included coagulopathy and rhabdomyolysis. Critical care nursing expertise in early assessment and intervention was integral to survival for this patient. The third case is of an elderly woman with an extensive medical history who experienced repeated prosthetic aortic graft infections, skin breakdown, and poor wound healing. The highlighted research is on low air loss beds. The critical care nurse made a significant difference in increasing each of these patient's chances for a successful hospital outcome. PMID- 7493263 TI - Aortic dissection: pathophysiology, diagnosis, and acute care management. AB - Acute aortic dissection is a life-threatening illness requiring early diagnosis and treatment. Management of this disease incorporates not only emergency department personnel, but also the medical, surgical, and intensive care specialties. If left untreated, the mortality rate of acute aortic disease exceeds 50% within 48 hours and 80% within 2 weeks, with a 5-year survival rate of 19%. The most common cause of death in untreated acute aortic dissection, regardless of etiology, is aortic rupture. In this article, the author reviews: the incidence and pathology; etiology; clinical manifestations; diagnostic workup; patient care management. PMID- 7493265 TI - Peripheral vascular assessment: finding your way through arteries and veins. AB - Vascular disease is a widespread health problem. Despite the advent of new, noninvasive and invasive testing methods, a thorough health history and physical examination remain the primary means of diagnosis. In this article, the author discusses vascular assessment of acute and critically ill patients, including inspection, palpation, and auscultation. Specific assessments for arterial disease and venous disease are discussed. A thorough vascular assessment aids in the planning and implementation of care and evaluation of response to treatment for these patients. PMID- 7493264 TI - Pain, pills, and possibilities: drug therapy in peripheral vascular disease. AB - Currently, pharmacologic therapy offers no cures for the patient with vascular disease. Drugs are available, however, to prevent or delay the end organ damage of vascular disease, to keep vessels patent as long as possible, and to provide some relief from the pain associated with vascular disease and cell damage. In this article, the author reviews the drugs used to manage hypertension- diuretics, beta blockers, alpha adrenergic blockers, angiotensin-converting enzyme inhibitors, calcium channel blockers, antiadrenergic drugs, and nitroprusside; and drugs used to maintain vessel patency in peripheral vascular disease--peripheral vasodilators and anticoagulants; and reviews one regimen for dealing with the chronic pain of vascular disease--narcotics and tricyclic antidepressants. PMID- 7493266 TI - Unraveling the mystery: clues to systemic vasculitic disorders. AB - In this article, the author reviews some of the most common vasculitic disorders, including: polyarteritis nodosa, Churg-Strauss syndrome, Wegener's granulomatosis, hypersensitivity vasculitis, Henoch-Schonlein purpura, giant cell arteritis, and Takayasu's arteritis. Information about the definition, general considerations, clinical findings, diagnostic tests, classification criteria, and treatment is included. The significance of early recognition of these disorders in the critical care area is discussed. PMID- 7493267 TI - Systemically induced vasculitis in children. AB - Vascular diseases can be quite grave in children. Common diseases in children that cause a generalized vasculitis include Kawasaki disease, Henoch-Schonlein purpura, and Rocky Mountain spotted fever. Rheumatic fever, though preventable, is increasing in incidence and may cause arteritis, and therefore is discussed as a vascular disease. Developmental considerations should be considered when providing adequate care. Play therapy is a crucial part of every day in a child's life, even when ill with a vascular illness. Play helps a child gain control over the stress of a serious illness and hospitalization. PMID- 7493269 TI - [Nursing research in intensive care]. PMID- 7493268 TI - Peripheral vascular disease: educational concerns for patients with a chronic disease in a changing health-care environment. AB - Education about vascular disease requires information to change habits related to diet, activity, and personal health. Uncertainty significantly impacts the ability of individuals with a chronic disease to feel secure in treatment programs. A clear understanding of the disease, mutual development of measurable goals, clear communication, and a commitment to change provide the format for a successful education and prevention program. As the health-care system expects each individual to assume more responsibility for health promotion and maintenance, education will be key. PMID- 7493270 TI - [Nursing records]. AB - The good evolution of intensive care demands an intense and fluent communication. It is important for the attention to patients as well as for a better exploitation of the experience obtained day after day to have a written register of all the data, results, judgements and relevant actions which are being performed. PMID- 7493271 TI - [Intrahospital transportation of critically ill children]. AB - The intrahospital transport of a critically ill child is always a risky procedure. While it is being done, some complications which can worsen their initial situation may appear, so the benefits that this transport can provide must outweigh the possible risks. Preparing the patient and succeeding in performing a safe transport need the use of the proper equipment and human resources. Its degree of complexity and preparation must be proportional to the situation of instability of the patient and to the probability of increasing such instability, which implies performing a careful evaluation of the child and its real and potential needs previously. We also state some general ideas about the way of preparing and performing this kind of intrahospital transport in its different stages. PMID- 7493272 TI - [Drugs in cardiology. I. Digitalis glycosides]. PMID- 7493273 TI - [Description of the intensive care unit at the Hospital San Millan, Logrono (La Rioja)]. PMID- 7493274 TI - [Intracranial hypertension in brain injuries]. AB - We review the physiopathology of the intracraneal hypertension (ICH), the present device for its monitoring and the relevance of the PIC waves from some significant figures of the epidemiology of traffic accidents, which are the first cause of the craneoencephalical traumatisms (CET) which are admitted into an Intensive Care Unit. Following that we examine the valuing criteria of the patients with intracraneal hypertension, their evolution through their symptoms, the medical treatment, and the nursing actions which can alter the PIC. PMID- 7493275 TI - [Consuelo Zazpe Oyarzun]. PMID- 7493276 TI - [The professional nursing career, a future alternative]. PMID- 7493277 TI - [Veno-venous continuous hemodialysis: implications for nurses]. AB - Veno-venous haemodialysis is an extra-renal depuration method of recent application in our Unit which adds the advantage of blood impulsion by bomb, which provides a higher control over the volume of ultrafilterring and decreases the possible haemodynamic alterations. In this work we show our experience about the incorporation of this new process, the important role that the nursing staff acquire in the circuit handling as well as in patients care. To do so, between March and October 1993 we performed a follow-up of this technique in 11 cases collecting data about complications with the circuit and patient improvement, circuit maintainance, reasons for change, volumes and balances obtained. PMID- 7493278 TI - [Classification of written nursing records in the intensive care unit of the Alicante General Hospital]. AB - During the last few years the need to implant written Nursing registers has been transferred from Schools to the different official organisms. The Servicio Valenciano de Salud took it as an aim to achieve by 1992 and we also know the Insalud and other autonomic sanitary organisms have shown their interest for the creation of such registers, and there is also a general consensus at a theoretical level of the requirements they ought to fulfill. With this work we plan to: first, know the percentage of nurses who, in our unit, write intershift reports. Second, know the kind of event or activities which we write down in these intershift reports, and last, classify the contents of such registers. To do this we analyse 113 intershift reports, randomly chosen and without previous notice to the unit staff, of the total number of available intershift reports in different periods of time between January and November 1993. A content analysis was performed with a result of 13 categories. The later classification of the contents of the registers in this category item, by equivalence among judges, indicated the high reliability of the process. The Agreement Degree obtained was 98.08% for all the categories. The results obtained indicate: 1) only medical diagnosis are used, not nursing ones 2) the events written down are of a biomedical type 3) information from graphics is repeated 4) the percentage of written intershift reports was 94.95%. Several questions arisen from the results obtained and the bibliography used for this research are revised. PMID- 7493279 TI - [Design of a program of quality control of nursing in an intensive care unit]. AB - Although in the last few years care quality programs have started to be used in services, these are still few in nursing equipment. Our aim is to show the process we have followed to design a nursing care quality program in ICU. We developed the stages of process, where a list of general activities performed in the service was created to establish a priority later, as it was necessary, due to the high number of aspects or activities to monitor, to choose those we would use to start the quality control program. The ultimate order was settled with the following activities: 1. Surveillance of the infection. 2. Accidental withdrawal of tubes and catheters. 3. Pressure ulcers. PMID- 7493280 TI - [Analysis and handling of the jugular catheter in the determination of cerebral anoxia]. AB - The increase in the incidence of Craneoencephalic Trauma takes us to the search of new techniques such as the calculus of the Fraction of Oxygen Brain Extraction, determined by the channelling of the jugular bulb and later extraction of cerebral venous gasometries simultaneously to peripheric arterial gasometries. The aim of our work is to let Nursing know its handling as well as the diagnostic and therapeutical advantages and their associated complications. We conclude that this technique shows important advantages in the handling of endocraneal hypertension without being a work overload for Nursing. We suggest a performing guide to avoid the most common complications during its maintainance. PMID- 7493281 TI - [Drugs in cardiology. Angiotensin-converting enzyme inhibitors]. PMID- 7493282 TI - [The intensive care unit of the Germans Trias i Pujol Hospital]. PMID- 7493283 TI - [Health in Cuba]. PMID- 7493284 TI - [From the other side of the mirror]. PMID- 7493285 TI - [Analysis of the situation of intensive care units]. AB - Since 1990, the Sociedad Espanola de Enfermeria Intensiva y Unidades Coronarias has carried out initiatives aiming to collect data which allows us to make an analysis of the status of the ICU nurses and the characteristics of the Units where these professionals develop their activity. This project was performed during 1992 and 1993, with a survey in 132 sanitary centres of the public network in the national territory, in which information about the number of ICUs and their number of beds, number of nurses and assistants in the institution and those ones specifically assigned to ICU, number of admissions, average stay, occupation percentage, admission causes, use of systems of valuation of the seriousness index, staff seniority, shift systems, staff stability, etc; all this data and others referring to 1991. From 53 hospitalary centres that answered the questionnaire, we obtained information of 94 ICUs in which there was an average of 10.4 beds, with 550 admissions per unit (average/year) and an index of occupation of 78%. The average number of nurses who work in each ICU was 22.7, with a nurse/bed ratio in the global calculation of staff of 2.18. In the analysis of shifts, the nurse/bed ratio was one nurse to every 2.08 patients (1:2.08) when there is a maximum of staff during the morning shift. This index is lower in the other shifts. The average of assistants is 12.1 with an assistant/bed ratio of 1.17. When studying the shifts systems, the rotatory shift outstands in 53.19% of ICUs and the existence of rotation systems of staff in other units is 8.5%. With reference to the characteristics of the staff, the average seniority of Nurses was 6.7 years, 76.4% have own their post and the percentage of new intake in 1991 was 22.5%. The lack of incentives to work in ICU is notable, the most problematic aspects being the insufficient economic remuneration, lack of motivation, scarce human resources and insufficient training, among others. Finally, according to the teaching and research activity performed in the units, 5.4% of ICUs have a nurse in the unit with exclusive dedication to teaching, in 20.4% there are nurses who keep a contractual relationship with University Nursing Schools, in 42.6% of the units there are training programs for the new staff and 67% perform some sort of research activity although in most of the cases it is only sporadic. PMID- 7493288 TI - [Description of the intensive care unit at the General Hospital Miramar in Palma de Mallorca]. PMID- 7493286 TI - [Intrahospital transportation of critical patients]. AB - Critically ill patients often need to be transferred for a short period of time for diagnostical or therapeutical reasons to other areas outside the intensive care unit which are less safe than their own unit and suppose a potential risk of deterioration in the patient's status. We analyse prospectively the intrahospitalary transfer in 50 patients and study the hemodynamic, ventilatory and neurological variations before and after the transfer. 93.7% of our patients were transferred for diagnostical reasons, basically to the radiodiagnosis service (85.4% for TAC performance), only 6.25% were transferred for therapeutical reasons, all of them to the operating theatre. All the patients included in the study were subjected to: -mechanic ventilation, electrocardiographic monitoring (ECG), invasive arterial monitoring (TA), monitoring of arterial saturation of O2 using pulsioximetry, drugs infusion through volumetric bombs and intracraneal pressure monitoring through intra ventricular catheter (in 18 cases). The intrahospitalary transfer was performed with: -Portable ventilator, ECG monitoring, TA, PIC and pulsioximetry. Before and after the transfer different parameters were registered: -Inspiratory fraction of O2 (FiO2), TA, cardiac frequency, PIC, arterial gasometry (pH, PAO2, PACO2). There were no complications in any of the cases, the gasometric alterations were due to the change of respiratory parameters for the transfer (increase of the FiO2 and prophylactic ventilation in all the cases). We recommend: -Use of the portable ventilator, volumetric bombs, hemodynamic monitoring and uninterrupted pulsioximetry and the presence of qualified staff (doctor and ICU nurse) during the transfer. PMID- 7493287 TI - [Beta blockers]. PMID- 7493289 TI - [Five years as the president of the Spanish Society of Intensive care and Coronary Units]. PMID- 7493290 TI - Approaches to the development of novel inhibitors of hepatitis C virus replication. AB - Chronic infection with the hepatitis C virus is a major health problem. Conventional therapy, with interferon-alpha is effective in only a minority of patients. The failure of current treatments has led to a major initiative to identify new antiviral agents. In the absence of a tissue culture model for hepatitis C infection the pharmaceutical industry has been obliged to investigate the basic biology of hepatitis C viral replication. Such studies have identified novel translational regulatory elements and new proteolytic enzymes which may serve as targets for new antiviral drugs. PMID- 7493291 TI - Precore and X region mutants in hepatitis B virus infections among renal dialysis patients. AB - Hepatitis B virus (HBV) variants containing mutations within the X and the precore regions of the viral genome were demonstrated by polymerase chain reaction (PCR) amplification and DNA sequencing in renal dialysis patients with different serological patterns of HBV infection. Among carriers, X region deletion mutants predominated in patients who lost hepatitis B e antigen (HBeAg), or developed anti-HBe, but not in persistently HBeAg-positive patients. The precore region remained wild type in all carriers whether or not they seroconverted from HBeAg to anti-HBe. The frequency of precore and X region mutants was greatest among non-carrier patients with viral antibodies as the only indication of infection and among patients with non-A, non-B hepatitis (NANBH), suggesting an inverse relationship between the presence of wild type HBV markers and the presence of HBV mutants. Furthermore, the detection of one but not the other mutation in many serum samples suggests that these mutations are independently selected for during infection. Finally, the absence of HBV DNA in 21 'uninfected' dialysis patients with normal transaminases and no viral serology, suggests that replication of these mutants is associated with hepatitis. These results have important implications for HBV screening and treatment, as well as for the pathogenesis of chronic infection. PMID- 7493292 TI - Distribution of three major hepatitis C virus genotypes in Italy. A multicentre study of 495 patients with chronic hepatitis C. AB - Different genotypes of hepatitis C virus (HCV) have been shown to have distinct geographical distribution and to associate with variable clinical features. To evaluate their role in chronic hepatitis in Italian patients, we studied 495 consecutive cases with chronic hepatitis C seen in nine sentinel centres homogeneously distributed over Italy. HCV genotyping was carried out using a dot blot hybridization assay with genotype-specific probes. Four hundred and eleven patients were viraemic and could be evaluated: 57% were found to be infected with HCV-1, 31% with HCV-2, 8% with HCV-3, 1% showed mixed infection and 3% were ascribed to HCV-2b or HCV-4 by direct sequencing. Geographical distribution showed discrete territorial variations. A history of drug addiction was commoner in patients infected with HCV-3. There were no significant differences in activity of liver disease among different HCV genotypes but the response to interferon therapy was reduced in patients infected with HCV-1 compared to HCV-2 or HCV-3. PMID- 7493293 TI - Coinfection with hepatitis B and C or B, C and delta viruses results in severe chronic liver disease and responds poorly to interferon-alpha treatment. AB - Chronic coinfection with the hepatitis B (HBV) and hepatitis delta (HDV) viruses is known to cause severe liver disease, but the importance of coinfection with hepatitis C virus (HCV) and HBV has not been well documented. In the present study, the clinical and pathological severity of liver disease among patients with hepatitis resulting from multiple viruses was examined and an open trial of the efficacy of interferon-alpha 2b (IFN-alpha) treatment was conducted. Nineteen patients with chronic HBV and HCV infection and 17 with HBV, HCV and HDV infection were studied; 12 in each group underwent liver biopsy. For each coinfected patient, two patients infected with HCV alone were selected as controls, and these were matched for age and risk factor and were estimated to have been infected for a similar duration. Coinfection with HBV and HCV or HBV, HCV and HDV was associated with more severe liver disease than HCV alone (P < 0.01); total Scheuer score, portal and lobular inflammation and fibrosis were all worse in coinfected subjects. Eight patients with chronic HBV and HCV were treated with recombinant IFN-alpha 2b [3 million units (MU), thrice weekly for 6 months]. Liver function tests normalized in two patients and one lost hepatitis B surface antigen (HBsAg). Seven patients with hepatitis B, C and delta coinfection were treated with the same regimen and only one normalized serum alanine aminotransferase (ALT) during (and after) treatment. It is concluded that coinfection with multiple hepatitis viruses is associated with histologically more severe liver disease than HCV alone.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7493294 TI - Hepatitis C virus infection in the asymptomatic British blood donor. AB - Blood donor screening for hepatitis C virus (HCV) antibodies is now routine. Most blood transfusion services recommend that seropositive donors are referred for further investigation. Southern European studies suggest that many asymptomatic seropositive donors have clinically significant liver disease. Seropositive donors in areas of high prevalence may not, however, be representative of British donors. We have prospectively examined the prevalence and severity of HCV infection in a British volunteer blood donor population. During a 14 month period, only 0.35% (999/287,332) of all donors in the West Midlands were anti-HCV (screening assay) positive. Only 5% (52/999) of these were confirmed true seropositive. Nearly 80% (41/52) of seropositive donors were referred to the Queen Elizabeth Hospital Liver Unit for further investigation. Most underwent complete investigation, including liver biopsy. Forty of forty-one donors had biochemical, histological, or virological evidence of persistent viral infection. Histological changes were generally mild and none was cirrhotic. Covertly infected patients had less severe disease than those with an overt risk factor for HCV exposure. In the British Midlands, the prevalence of blood donor seropositivity is low. In contrast with seropositive Southern European donors, the British donor is more likely to belong to an at-risk group for parenteral exposure and is less likely to have severe histological changes. This study highlights the importance of developing locally relevant guidelines for the counselling and investigation of anti-HCV-positive blood donors. PMID- 7493295 TI - Comparison of two second-generation anti-hepatitis C virus ELISA on 21431 US blood donor samples. AB - We have compared two different second-generation (2.0) enzyme-linked immunosorbent assays (ELISA) for the presence of antibodies to hepatitis C virus (anti-HCV) in blood from volunteer, unpaid donors. At two separate blood centres, a total of 21,431 donor samples were tested with Abbott Anti-HCV 2.0 ELISA and Ortho Anti-HCV 2.0 ELISA. Samples found to be repeatedly reactive were tested by supplemental/investigational assays. MATRIX HCV (Abbott) and anti-HCV RIBA II (Ortho/Chiron), to 'confirm' the presence of anti-HCV. Discordant ELISA samples were additionally tested by the polymerase chain reaction (PCR) for the presence of HCV RNA. The Abbott anti-HCV assay had a repeatedly reactive rate of 0.59% (127/21,431) and the Ortho anti-HCV assay 0.51% (110/21,431). Overall agreement between assays was 99.76%, 72/127 (56.7%) of Abbott repeatedly reactive samples confirmed on MATRIX and 61/127 (48.0%) on RIBAII; 70/110 (63.6%) of Ortho repeatedly reactivate samples confirmed on MATRIX and 61/110 (55.5%) on RIBA II. Discordant ELISA samples tested by PCR yielded negative results. Hence the two ELISA had equal sensitivity, as defined by detection of true positive samples; the slightly lower specificity of the Abbott Anti-HCV 2.0 ELISA may be owing to culling of donors with a false positive test by Ortho's Anti-HCV 1.0 and 2.0 ELISA tests (the routine tests in place at each blood centre). A sample found to be repeatedly reactive by two different ELISA tests for anti-HCV is likely to be a true positive and may not require further 'confirmatory' testing. PMID- 7493298 TI - Detection and quantitation of hepatitis B virus DNA: comparison of two commercial hybridization assays with polymerase chain reaction. AB - Serum hepatitis B virus (HBV) DNA is now the most important and reliable marker for monitoring hepatitis B viral replication. Quantitative detection of HBV DNA in serum is based on commercial standardized molecular hybridization test systems. We compared two hybridization assays, the Digene Hybrid Capture assay (Digene Diagnostics, Beltsville, MD) and the Abbott HBV DNA assay (Abbott Laboratories, North Chicago, IL, USA) with the polymerase chain reaction (PCR) technique, for detection and quantitative measurement of serum HBV DNA. Forty-two patients with various HBV serological marker profiles were included in this study. The patients were divided into four groups according to their HBV DNA values after HBV DNA determination in the serum by the Abbott assay. For each patient HBV DNA was then determined by the Digene assay and by PCR. In the case of Digene and PCR there was a 97.6% correspondence in the outcome of the two methods, whereas in the Abbott assay and PCR there was only 69% correspondence. The McNemar test of symmetry showed no statistically significant difference between the Digene assay and PCR, whereas there was a significant difference (P < 0.01) in the Abbott assay and PCR. For low positive HBV DNA values between 1.5 and 20 pg ml-1 the Abbott assay yields inconclusive results. Differences observed between the two hybridization assays underline the need for standardization of HBV DNA quantitation. PMID- 7493296 TI - Genetic control of the murine humoral response to distinct epitopes of hepatitis C virus core protein. AB - Recombinant hepatitis C virus (HCV) core protein from aa1-164, designated cp1-10, was used to immunize mice. Antibodies to cp1-10 were produced in all seven strains of congenic mice; none of the strains could be considered low responders relative to the others. The mouse response against individual epitopes of HCV core protein varied from one strain to another: B10.RIII (H-2r) recognized all three peptides aa13-30, aa77-90, aa129-145; B10.D2 (H-2d), B10 (H-2b) and C3H.SW (H-2b) responded to aa13-30, aa77-90; B10.M (H-2f), B10.BR (H-2k) and C3H/Hej (H 2k) reacted with aa13-30 only. Competitive inhibition of binding demonstrated that antibody to the peptide was inhibited by cp1-10 protein and the corresponding peptide only. Recombinant HCV core protein is highly immunogenic and can elicit good antibody response in mice. The aa13-30 is a major epitope of HCV core protein in mice. The humoral response to the distinct epitopes was regulated by the H-2 genes. Further analysis indicated that the I-a locus of H-2 genes determined the antibody response to aa13-30 and 77-90. These results suggest that the variation of antibody responses to HCV in humans may partially contribute to different outcomes of HCV infection. PMID- 7493297 TI - Clinical efficacy of intramuscular human interferon-beta vs interferon-alpha 2b for the treatment of chronic hepatitis C. AB - We have conducted a randomized study to compare the efficacy and tolerance of human interferon (IFN) beta vs recombinant IFN-alpha 2b in patients with chronic active hepatitis C. Forty patients were included: 21 received IFN-alpha (group A) and 19 IFN-beta (group B). IFN was administered intramuscularly at a dose of 6 MU three times a week (tiw) for 2 months (induction phase), followed by 3 MU tiw for 4 months. Clinical, epidemiological and pathological features were similar in the two groups. Normal alanine aminotransferase (ALT) values at the end of treatment was regarded as a response to therapy and the response rate was 57% (12/21) in group A and 5.2% (1/19) in group B (P < 0.01). Both types of IFN induced a significant decrease in mean ALT values by the end of the induction phase (P < 0.01). When the dose was reduced to 3 MU, a marked, but not significant increase in ALT, was seen in group B, whereas no increase was seen in group A. IFN-beta was better tolerated and haematological adverse effects (platelet and leucocyte decrease) were less pronounced with IFN-beta. Hence, human IFN-beta was less effective than IFN-alpha in treating chronic hepatitis C virus (HCV). Doses of IFN-beta of 3 MU intramuscular (IM) tiw were clearly insufficient and it remains to be established whether higher doses of intramuscularly IFN-beta can be useful. PMID- 7493299 TI - Liver transplantation for acute and chronic viral hepatitis. AB - Hepatotrophic viruses are responsible for a substantial proportion of cases of both end-stage chronic liver disease and of acute liver failure which are treated by liver transplantation. We review here current practice in transplantation for viral-induced liver disease addressing, in particular, the selection of patients for transplantation and the increasingly recognized problem of recurrent disease in liver grafts. PMID- 7493300 TI - Long-term titrated recombinant interferon-alpha 2a in chronic hepatitis C: a randomized controlled trial. AB - The efficacy and tolerability of 12-month treatment with titrated doses of recombinant interferon-alpha 2a (IFN-alpha 2a) in chronic hepatitis C were studied in 67 consecutively recruited patients randomly assigned either to a starting dose of IFN-alpha 2a 6 MU, subsequently adjusted to the serum alanine aminotransferase (ALT) response (n = 35), or to no therapy (n = 32; controls). End-of-treatment ALT levels were normal and hepatitis C virus (HCV) RNA was negative by nested polymerase chain reaction (PCR) in 17 (49%) treated patients compared to none of the controls (P < 0.001). During the 12 months after stopping treatment the number of patients who remained in remission was eight (23%) and one respectively (4%) (P = 0.031). Follow-up liver biopsy showed reduced hepatic inflammation in 80% of treated patients and in 29% of controls (P < 0.001). The eight sustained responders and 27 non-responders or relapsers received similar mean total doses of IFN (565 MU vs 545 MU) and had a similar incidence of anti IFN neutralizing antibodys (13% vs 19%). Absence of cirrhosis was the only independent pretreatment parameter that predicted a sustained response. In conclusion, a mean cumulative dose of IFN 549 MU, titrated over 12 months, was well tolerated, and resulted in the long-term clearance of HCV RNA and normal ALT levels in 23% of patients. PMID- 7493301 TI - Hepatitis B e antigen negative chronic active hepatitis: hepatitis B virus core mutations occur predominantly in known antigenic determinants. AB - In chronic hepatitis B virus (HBV) infection seroconversion from hepatitis B e antigen (HBeAg) to hepatitis B e antibody (HBeAb) may be followed either by remission of the disease with low-level viraemia, or by continuing inflammation with high-level viraemia. In both situations the virus may acquire a mutation in the precore sequence which prevents it from encoding HBeAg. We now show that the number of amino acid substitutions in the HBV core is low in viral sequences from patients with HBeAg positive chronic liver disease and HBeAg negative HBeAb positive patients in remission, but the frequency of substitutions is high in HBeAg, negative HBeAb positive patients with active liver disease. Furthermore we show that these substitutions cluster in the promiscuous CD4+ T-helper-cell epitope and in HBV core/e antibody binding determinants, but are not found in regions recognized by major histocompatability complex (MHC) restricted cytotoxic T lymphocytes. Sequential viral sequences from patients before and after HBeAg/HbeAb seroconversion shows that core mutations arise either at the same time or after the precore stop mutation which prevents the virus from encoding HBeAg. These results are consistent with the hypothesis that after clearance of HBeAg, mutations in regions of the virus recognized by CD4+ helper T cells and B cells allow persistence of the HBe negative virus in HBeAb positive patients with viraemia and active hepatitis. PMID- 7493302 TI - Inhibition of hepatitis B viral gene expression by antisense phosphorothioate oligodeoxynucleotides. AB - The inhibitory effect of antisense phosphorothioate oligodeoxyribonucleotides (S oligos) on the production of hepatitis B surface antigen (HBsAg) and hepatitis B e antigen (HBeAg) was studied using Hep G2 cells transfected with hepatitis B virus (HBV). Synthetic antisense oligos (15-mers) directed against the cap site of mRNA transcribed from the SP II promoter and regions of the translational initiation site of the S gene showed a sequence-specific, dose-dependent inhibitory effect on HBV gene expression between concentrations of 1.0 microM-5.0 microM. Oligos directed against the middle of the S gene had little effect on HBsAg and HBeAg expression, as did non-complementary random-sequence control. The cells remained viable throughout the experiments and no morphological abnormalities were observed with antisense S-oligos at concentrations below 20.0 microM. These results suggest a therapeutic potential for antisense oligonucleotides in the treatment of patients who are chronically infected with HBV. PMID- 7493303 TI - Predictors of sustained response, relapse and no response in patients with chronic hepatitis C treated with interferon-alpha. AB - Three main patterns of response are seen when interferon-alpha (IFN-alpha) is used for the treatment of chronic hepatitis C: 1 sustained response with alanine aminotransferase (ALT) normalization that is maintained after cessation of therapy, with or without clearance of serum hepatitis C virus (HCV) RNA; 2 transient response with ALT normalization during therapy followed by relapse after its withdrawal, and 3 no response with no or only partial reduction in ALT levels. In order to define variables that could predict each of these three types of response we studied 321 cases of chronic hepatitis C treated with IFN-alpha in two consecutive trials conducted in our Unit. By univariate analysis, age < 45 years (P < 0.01), known disease duration < 60 months (P < 0.01), normal gamma glutamyl-transpeptidase (gamma GT) levels (P < 0.01) and infection by HCV genotype 2 or HCV genotype 3 (P < 0.01) were found to be statistically associated with sustained response while age > 45 years (P < 0.01), body weight (P = 0.05), cirrhosis (P < 0.01) and elevated gamma GT levels (P < 0.01) were associated with no response. By multivariate analysis sustained response was predicted by HCV genotype 2 (P < 0.01) and HCV genotype 3 (P < 0.01), known disease duration (P < 0.01), patient's age (P < 0.05) and associated with the use of a more aggressive treatment schedule (P < 0.05). Transient response with relapse was predicted by known duration of disease (P < 0.05), HCV genotype 1 (P < 0.05) and female sex (P < 0.05). No response was statistically associated with elevated gamma GT levels (P < 0.01), higher body weight (P < 0.05) and with the less aggressive regimen of 3 MU of natural IFN-alpha given three times weekly for 6 months (P < 0.05). These results indicate that the HCV genotype as well as the schedule of treatment greatly affect the pattern of response to IFN in chronic hepatitis C and allow us to define criteria to predict which type of response is more likely in individual patients. PMID- 7493305 TI - Methods of transmission of hepatitis C. AB - The route of transmission of hepatitis C virus is still controversial. Parenteral exposure via blood or blood products leads to infection in the majority of cases, and the majority of intravenous drug users become infected by repetitive exposure to contaminated injection equipment. The risk of infection from a single needlestick injury is 5-15% and may depend on the size of the innoculum. Other parenteral routes of transmission may include traditional healing practices and the use of contaminated medical equipment. Transmission is less common within a family but the prevalence of hepatitis C viral antibodies is higher in family members and sexual partners of carriers than in the general population. There are some well-documented instances of acute hepatitis C occurring after a defined sexual exposure. Vertical transmission is rare unless the mother has high levels of circulating HCV RNA as may occur in those also infected with HIV. The detection of hepatitis C in saliva and the higher than expected prevalence of infection in dentists may point to the possibility of transmission by salivary contamination. There remain large numbers of hepatitis C carriers in whom no route of infection can be identified. PMID- 7493304 TI - Hepatitis C virus RNA and long-term response to recombinant interferon-alpha 2b in patients with chronic hepatitis C. AB - The effectiveness of recombinant interferon-alpha 2b (rIFN-alpha 2b) in eradicating hepatitis C virus (HCV) RNA from serum has not been completely assessed. We studied 39 patients with compensated chronic hepatitis C diagnosed by liver biopsy and positive HCV RNA measured by polymerase chain reaction (PCR). Group I consisted of 26 patients treated with 3 MU of rIFN-alpha 2b for 6 months; group II, 13 control patients observed for six months; and group III, 12 out of 13 patients from group II who subsequently received 5 MU of rIFN-alpha 2b for 6 months. In group I, 11 out of 23 (47.8%) patients who completed treatment had an immediate response and five (21.7%) had a sustained response to therapy six months after treatment. No response was observed in patients from group II. In group III, 7 out of 12 (58.3%) patients who completed treatment had an immediate response and none had a sustained response. Considering all patients who completed rIFN-alpha 2b treatment, HCV RNA remained positive at the end of therapy in three of five sustained responders (60%), six of 13 patients who relapsed (46.1%), and in all nonresponders (100%). HCV RNA was positive 6 months after therapy in four (80%), 13 (100%), and 17 (100%) patients respectively. All patients with a sustained response had normal aminotransferase levels 18 months after therapy. We conclude that in chronic hepatitis C rIFN-alpha 2b causes a significant immediate response but this is not sustained, only 2.8% of treated patients had a sustained loss of HCV RNA. Normal aminotransferase persist in the long term, despite persistence of HCV RNA. PMID- 7493306 TI - Evaluation of branched DNA signal amplification for the detection of hepatitis C virus RNA. AB - There is an increasing need for a practical assay to measure HCV RNA to assess the viral burden in chronic hepatitis C virus (HCV) infection as viral load relates to transmission and therapeutic response. This study evaluates branched DNA (bDNA) signal amplification, a technique that avoids many of the pitfalls of polymerase chain reaction (PCR). The bDNA assay uses a microtitre well format and a series of capture, target and amplification probes that bind RNA to the well and then successively bind oligonucleotides to the RNA and branched DNA molecules to the oligonucleotides. Enzyme-labelled probes are bound to the arms of the bDNA and light output from a chemiluminescent substrate is directly proportional to the amount of starting HCV RNA. Appropriate standards provide direct quantitation. Whereas PCR amplifies the HCV genome, bDNA amplifies the hybridization signal. In testing a standardized, coded panel, bDNA showed 100% specificity and detected five of six sera proven to transmit hepatitis C to the chimpanzee; PCR detected all six infectious sera. Serial samples were measured in two acute and five chronic cases of transfusion-associated hepatitis and in three commercial seroconversion panels. In acute cases, 10(7)-10(8) molecular equivalents per ml (eq per ml) of HCV RNA were detected prior to peak alanine aminotransferase (ALT) activity and then rapidly declined to non-detectable levels. Similar levels of HCV RNA were observed early in the course of two patients who progressed to chronic hepatitis; the chronic course was characterized by diminished, fluctuating and sometimes non-detectable levels of HCV RNA. In two chronic cases, HCV RNA was not detected, or only transiently detected by bDNA, but was present when assayed by PCR. In one chronic case, the periodicity of HCV RNA levels closely paralleled the fluctuations of ALT suggesting a relationship between viral replication and subsequent hepatocellular injury. In testing 50 blood donors whose anti-HCV reactivity was confirmed by a recombinant immunoblot assay (RIBA), HCV RNA was detected by bDNA in 41 (81%), while PCR was positive in 45 (90%); the overall concordance between bDNA and PCR in 100 anti-HCV enzyme immunoassays (EIA) reactive donor samples was 96%.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 7493307 TI - Post-hepatitis syndrome revisited. AB - To examine the role of acute hepatitis A and B infection in the aetiology of chronic fatigue syndrome and psychiatric morbidity we studied 40 patients with acute viral hepatitis A or B consecutively admitted to an infectious diseases unit and studied at least 6 months after recovery. Liver function tests (LFT) had returned to normal in each case. Forty-seven patients with other infectious diseases, of which 12 were presumed viral, admitted immediately after each hepatitis patient during the same period acted as controls. The main outcome measures were scores on a fatigue and muscle pain questionnaire, general health questionnaire (GHQ-12) and supplementary questions. The hepatitis cases scored significantly higher fatigue scores, GHQ-12 scores and muscle pain scores. Length of time since recovery from illness, age and sex were not confounding factors. Hepatitis cases were also less energetic, had greater weight change, had altered alcohol tolerance, had less exercise tolerance and felt less fit than the control group and compared with their premorbid state. Hence fatigue is more common after recovery in patients hospitalized for hepatitis A and B up to 30 months post infection compared with matched controls hospitalized for other infectious diseases. Hepatitis A and B infection is a risk factor for post-infection fatigue, intermittent fatigue, as well as for psychiatric morbidity. PMID- 7493308 TI - Ethnic differences in the prevalence of anti-hepatitis C antibodies and hepatitis B surface antigen in Israeli blood donors by age, sex, country of birth and origin. AB - The presence of anti-hepatitis C virus (HCV) antibodies frequently indicates both persistent infection and infectivity. Consequently, blood donors found to be anti HCV positive, are excluded from the donor pool. The aim of this study was to compare age, sex and ethnic differences in the prevalence of anti-HCV antibodies with that of hepatitis B surface antigen (HBsAg) among immigrant and Israeli-born blood donors. Anti-HCV antibodies were assayed by a second-generation enzyme immunoassay (EIA) and HBsAg by a standard EIA in a sample of 136,977 blood donors in Israel during 1992. The overall age-adjusted prevalence of anti-HCV antibodies was 0.66% in men and 0.55% in women, and for HBsAg, 0.85% and 0.44%, respectively. There was a significant increase in the prevalence of anti-HCV antibody with age, and significant differences by country of birth, with the highest prevalence found among those born in the former USSR and eastern Europe. This contrasted with the findings for the prevalence of HBsAg, where the highest rates were among those born in northern Africa. Among Israeli-born donors, differences in the prevalence of anti-HCV antibodies by parental country of origin were minimal and much less than for HBsAg. Hence the prevalence of anti HCV antibodies in Israel is strongly associated with age and country of birth but not with country of origin. There is little evidence of substantial vertical or intrafamilial transmission of HCV infection in Israel. PMID- 7493309 TI - Epidemiology of hepatitis A in Valencia, Spain: public health implications. AB - Between July 1988 and July 1989, sera from 1223 persons resident in the Valencia area of Spain were tested for antibodies against the hepatitis A virus. Sixty five per cent of serum samples were positive for anti-HAV (95% confidence interval = 62.4-67.6). The prevalence of anti-HAV increased significantly with age (odds ratio > 50 years = 69.8; 95% confidence interval = 26.5-183.4) and previous history of hepatitis A (odds ratio = 2.1; 95% confidence interval = 1.4 3.2). Prevalence decreased with higher educational level (odds ratio, university studies = 0.2; 95% confidence interval = 0.1-0.5). Overall, there has been a reduction of anti-HAV prevalence reflecting the decreasing exposure of the Spanish population to hepatitis A virus in recent years, particularly in the younger generations. The age of infection has increased, increasing the probability of future epidemics in groups previously protected by immunity acquired in early childhood. This new epidemiological pattern has strong public health implications, and universal childhood vaccination together with measures directed to improve sanitation may be the best public health strategy to protect the population. PMID- 7493310 TI - A study of hepatitis delta virus infection in patients with acute and chronic liver disease from northern India. AB - To assess the prevalence of hepatitis delta virus (H delta V) in northern India, 204 adult patients with acute and chronic liver disease who were positive for hepatitis B virus (HBV) markers were screened for anti-H delta V antibody by enzyme-linked immunosorbent assay (ELISA). Anti-H delta V antibodies were positive in 29 (14.2%) patients. The incidence of H delta V infection was higher (21.4%) in patients with chronic liver disease when compared with those with acute viral hepatitis (10.7%) (P < 0.05). H delta V antibodies were positive in 16.6% of patients with fulminant hepatic failure (FHF) and in 25% of cases with hepatocellular carcinoma. Co-infections were significantly higher in acute hepatitis (80%), while superinfections predominated (66.7%) in chronic liver disease (P < 0.05). Our data show that H delta V is endemic in northern India and should be considered a major health problem. PMID- 7493312 TI - Raised levels of antibodies to human viruses at the clinical onset of autoimmune chronic active hepatitis. AB - Patients with autoimmune chronic active hepatitis (AICAH) often have very high titres of antibodies to rubella and/or measles virus. In the present study a young girl at the clinical onset of AICAH exhibited very high titres of antibodies against influenza viruses A and B, parainfluenza viruses, rubella virus and varicella-zoster virus. The titres normalized over 2 months except for rubella and varicella-zoster antibodies. Strong reactivities were seen against the rubella structural proteins E1, E2 and C in Western blot but IgM antibodies were not demonstrated. Total IgG was increased with normal ratios of subclasses. The IgG1 was the dominant antibody to E1 and E2, while IgG4 dominated the anti-C response. There was no significant shift in subclass reactivities over one year from onset. The polymerase chain reaction (PCR), using a nested primer set, was negative for rubella virus RNA in a liver biopsy obtained at the clinical onset and in peripheral blood mononuclear cells (PBMC) 1 year later. Co-cultivation experiments using PBMC and permissive cell lines were also negative for rubella virus. Hence, in the very early phase of AICAH there may be a transiently enhanced antibody response to various unrelated viruses. PMID- 7493311 TI - Re-treatment of interferon-resistant patients with chronic hepatitis C with interferon-alpha. AB - Non-responders to 6-months treatment with recombinant interferon (rIFN)-alpha, 3 MU thrice weekly (primary non-responders) were treated for 6 further months with the same therapy or with a double dose of rIFN-alpha or with a different type of IFN (L-IFN). 112 primary non-responders were randomly enrolled into four groups of 28 patients each over a period of 4 years and were followed up for 6 months: group A continued the same dose of rIFN-alpha, group B was treated with the same rIFN-alpha but received a double dose (6 MU thrice weekly), group C received L IFN, 3 MU thrice weekly, and group D stopped IFN therapy and did not receive any treatment. Patients were examined at monthly intervals and response was defined as a complete normalization of alanine amino transferase (ALT). The four groups were homogeneous as to age, sex, duration of the disease, probable source of infection, histological diagnosis. ALT and gamma glutamyl transferase (gamma GT) levels. No patient discontinued therapy for side-effects. Further treatment with rIFN-alpha 3 MU thrice weekly (group A) induced normalization of ALT levels in four patients (14%); treatment with double-dosed rIFN-alpha (group B) induced normalization of liver enzymes in six cases (21%); a different type of interferon (L-IFN) (group C) achieved normalization of serum ALT in five patients (18%). None of 28 primary non-responders who did not receive any treatment (group D) showed normalization of ALT levels. None of the patients was anti-HCV negative at the end of the study and no statistically significant difference was noted between responders and non-responders to the second course of IFN therapy as to age, sex, duration of the disease. ALT and gamma GT levels at the end of the trial. Overall at the end of the study the primary non-responders with normal levels of ALT were 15/112 (13%), with a therapeutic advantage of 7%. No statistically significant difference in the response rate was found among patients who continued IFN therapy, but prolongation of rIFN-alpha treatment at double dosage seems to be the best therapeutic regimen. PMID- 7493313 TI - An important RNA tertiary interaction of group I and group II introns is implicated in gram-positive RNase P RNAs. PMID- 7493314 TI - In vivo structural analysis of spliced leader RNAs in Trypanosoma brucei and Leptomonas collosoma: a flexible structure that is independent of cap4 methylations. AB - The formation of the mRNA 5' end in trypanosomatid protozoa is carried out by trans-splicing, which transfers a spliced leader (SL) sequence and its hypermethylated cap (cap4) from the SL RNA to the pre-mRNA. Previous in vitro studies with synthetic uncapped RNAs have shown that the SL sequence of Leptomonas collosoma can assume two alternate conformations, Form 1 and Form 2, with Form 1 being the dominant one. To gain information about the structure of the SL RNA in vivo, in its protein-rich environment, we have used permeable Trypanosoma brucei and L. collosoma cells for chemical modification experiments. We introduce the use in vivo of the water-soluble reagents CMCT and kethoxal. In contrast to the in vitro results, the Form 2 secondary structure predominates. However, there are chemically accessible regions that suggest conformational flexibility in SL RNPs and a chemically inaccessible region suggestive of protection by protein or involvement in tertiary interactions. Using complementary 2'-O-methyl RNA oligonucleotides, we show that T. brucei SL RNA can be induced to switch conformation in vivo. SL RNA stripped of proteins and probed in vitro does not display the same Form 2 bias, indicating that SL RNA structure is determined, at least in part, by its RNP context. Finally, the methyl groups of the cap4 do not seem to affect the secondary structure of T. brucei SL RNA, as shown by chemical modification of undermethylated SL RNA probed in vivo. PMID- 7493315 TI - Analysis of the structure of Tetrahymena nuclear RNAs in vivo: telomerase RNA, the self-splicing rRNA intron, and U2 snRNA. AB - Dimethyl sulfate modification of RNA in living Tetrahymena thermophila allowed assessment of RNA secondary structure and protein association. The self-splicing rRNA intron had the same methylation pattern in vivo as in vitro, indicating that the structures are equivalent and suggesting that this RNA is not stably associated with protein in the nucleolus. Methylation was consistent with the current secondary structure model. Much of telomerase RNA was protected from methylation in vivo, but the A's and C's in the template region were very reactive. Thus, most telomerase is not base paired to telomeres in vivo. Protein free telomerase RNA adopts a structure different from that in vivo, especially in the template and pseudoknot regions. The U2 snRNA showed methylation protection at the Sm protein-binding sequence and the mRNA branch site recognition sequence. For both telomerase RNA and U2 snRNA, the in vivo methylation pattern corresponded much better to the structure determined by comparative sequence analysis than did the in vitro methylation pattern. Thus, as expected, comparative analysis gives the structure of the RNA in vivo. PMID- 7493316 TI - The final stages of spliceosome maturation require Spp2p that can interact with the DEAH box protein Prp2p and promote step 1 of splicing. AB - Pre-mRNA processing occurs by assembly of splicing factors on the substrate to form the spliceosome followed by two consecutive RNA cleavage-ligation reactions. The Prp2 protein hydrolyzes ATP and is required for the first reaction (Yean SL, Lin RJ, 1991, Mol Cell Biol 11:5571-5577; Kim SH, Smith J, Claude A, Lin RJ, 1992, EMBO J 11:2319-2326). The Saccharomyces cerevisiae SPP2 gene was previously identified as a high-copy suppressor of temperature-sensitive prp2 mutants (Last RL, Maddock JR, Woolford JL Jr, 1987, Genetics 117:619-631). We have characterized the function of Spp2p in vivo and in vitro. Spp2p is an essential protein required for the first RNA cleavage reaction in vivo. Depletion of Spp2p from yeast cells results in accumulation of unspliced pre-mRNAs. A temperature sensitive spp2-1 mutant accumulates pre-mRNAs in vivo and is unable to undergo the first splicing reaction in vitro. However, spliceosomal complexes are assembled in extracts prepared from the mutant. We show that Spp2p function is required after spliceosome assembly but prior to the first reaction. Spp2p associates with the spliceosome before the first RNA cleavage reaction and is likely to be released from the spliceosome following ATP hydrolysis by Prp2p. The Prp2 and Spp2 proteins are capable of physically interacting with each other. These results suggest that Spp2p interacts with Prp2p in the spliceosome prior to the first cleavage-ligation reaction. Spp2p is the first protein that has been found to interact with a DEAD/H box splicing factor. PMID- 7493317 TI - Branch-point attack in group II introns is a highly reversible transesterification, providing a potential proofreading mechanism for 5'-splice site selection. AB - By examining the first step of group II intron splicing in the absence of the second step, we have found that there is an interplay of three distinct reactions at the 5'-splice site: branching, reverse branching, and hydrolytic cleavage. This approach has yielded the first kinetic parameters describing eukaryotic branching and establishes that group II intron catalysis can proceed on a rapid timescale. The efficient reversibility of the first step is due to increased conformational organization in the branched intermediate and it has several important mechanistic implications. Reversibility in the first step requires that the second step of splicing serve as a kinetic trap, thus driving splicing to completion and coordinating the first and second step of splicing. Facile reverse branching also provides the intron with a proofreading mechanism to control the fidelity of 5'-splice site selection and it provides a kinetic basis for the apparent mobility of group II introns. PMID- 7493318 TI - Sequential recognition of the pre-mRNA branch point by U2AF65 and a novel spliceosome-associated 28-kDa protein. AB - Splicing of pre-mRNAs occurs via a lariat intermediate in which an intronic adenosine, embedded within a branch point sequence, forms a 2',5'-phosphodiester bond (RNA branch) with the 5' end of the intron. How the branch point is recognized and activated remains largely unknown. Using site-specific photochemical cross-linking, we have identified two proteins that specifically interact with the branch point during the splicing reaction. U2AF65, an essential splicing factor that binds to the adjacent polypyrimidine tract, crosslinks to the branch point at the earliest stage of spliceosome formation in an ATP independent manner. A novel 28-kDa protein, which is a constituent of the mature spliceosome, contacts the branch point after the first catalytic step. Our results indicate that the branch point is sequentially recognized by distinct splicing factors in the course of the splicing reaction. PMID- 7493319 TI - Transfer RNA aminoacylation: identification of a critical ribose 2'-hydroxyl-base interaction. AB - To understand the relationship between tRNA architecture and specific aminoacylation by aminoacyl-tRNA synthetases, we performed kinetic assays of Escherichia coli tRNA(Pro) molecules containing single deoxynucleotide substitutions. We identified an important 2'-hydroxyl group at position U8 (of 22 positions probed). Chemical modification studies showed that this 2'-hydroxyl interacts with either the N1 or the exocyclic amine of G46 in a hydrogen bonding interaction that contributes 1.8 kcal/mol to the free energy of activation for aminoacylation. Molecular modeling of tRNA(Pro) supports the existence of this interaction. This is the first study to identify a specific ribose 2'-hydroxyl base interaction in the core region of a tRNA molecule that makes a thermodynamically significant contribution to aminoacylation. PMID- 7493320 TI - RNA structural patterns and splicing: molecular basis for an RNA-based enhancer. AB - Efficient splicing of the 325-nt yeast (Saccharomyces cerevisiae) rp51b intron requires the presence of two short interacting sequences located 200 nt apart. We used the powerful technique of randomization-selection to probe the overall structure of the intron and to investigate its role in pre-mRNA splicing. We identified a number of alternative RNA-RNA interactions in the intron that promote efficient splicing, and we showed that similar base pairings can also improve splicing efficiency in artificially designed introns. Only a very limited amount of structural information is necessary to create or maintain such a mechanism. Our results suggest that the base pairing contributes transiently to the spliceosome assembly process, most likely by complementing interactions between splicing factors. We propose that splicing enhancement by structure represents a general mechanism operating in large yeast introns that evolutionarily preceded the protein-based splicing enhancers of higher eukaryotes. PMID- 7493321 TI - A dual-specificity pseudouridine synthase: an Escherichia coli synthase purified and cloned on the basis of its specificity for psi 746 in 23S RNA is also specific for psi 32 in tRNA(phe). AB - An Escherichia coli pseudouridine (psi) synthase, which forms both psi 746 in E. coli 23S ribosomal RNA and psi 32 in tRNA(Phe), has been isolated and cloned. The enzyme contains 219 amino acids and has a calculated MW of 24,432 Da. Amino acid sequence comparison with the three other psi synthases that have been cloned to date, two for tRNA and one for 16S RNA, did not reveal any common sequence motifs, despite the catalysis of a common reaction. The gene was cloned behind a (His)6 leader for affinity purification. Upon overexpression, most of the enzyme remained soluble in the cell cytoplasm and could be purified to homogeneity on a Ni(2+)-containing resin. The enzyme reacted with both full-length 23S RNA or a fragment from residues 1-847, forming 1 mol psi/mol RNA at position 746, a normal site for psi. The enzyme has no dependence on Mg2+. The same yield was obtained in 1 mM EDTA as in 10 mM Mg2+, and the rate was faster in EDTA than in Mg2+. Full length 16S RNA or fragments 1-526 or 1-678, as well as tRNA(Val) transcripts, were not modified in either EDTA or Mg2+. tRNA(Phe) transcripts, however, were modified with a yield of 1 mol psi/mol transcript at a rate in EDTA like that of 23S RNA. Sequencing showed all of the psi to be at position 32, a normal site for psi in this tRNA. Both 23S rRNA psi 746 and tRNA psi 32 occur in single-stranded segments of the same sequence, psi UGAAAA, closed by a stem. Therefore, this synthase may require for recognition only a short stretch of primary sequence 3' to the site of pseudouridylation. This is the first example of a dual-specificity modifying enzyme for RNA, that is, one which is specific for a single site in one RNA, and equally site-specific in a second class of RNA. The essentiality of these psi residues can now be assessed by disruption of the synthase gene. PMID- 7493322 TI - Finding splice sites within a wilderness of RNA. PMID- 7493323 TI - From stand-by to decoding site. Adjustment of the mRNA on the 30S ribosomal subunit under the influence of the initiation factors. AB - The hypothesis of an adjustment of the mRNA in its ribosomal channel under the influence of the initiation factors has been tested by site-directed crosslinking experiments. Complexes containing 30S subunits with bound mRNA having 4-thio uracil at specific positions were prepared in the presence or absence of initiation factors and/or fMet-tRNA and subjected to UV irradiation to obtain specific crosslinks of the radioactively labeled mRNA with bases of the 16S rRNA and with ribosomal proteins. The subsequent identification of the specific sites of both mRNA and rRNA and individual ribosomal proteins involved in the crosslinking, obtained under different conditions of complex formation, provide direct evidence for the occurrence of a partial relocation of the mRNA on the 30S ribosomal subunits under the influence of the factors. The nature of this mRNA relocation is compatible with our previous proposal of a shift of the template from an initial ribosomal "stand-by site" to a second site closer to that occupied when the initiation triplet of the mRNA is decoded in the P-site. PMID- 7493324 TI - Escherichia coli protein StpA stimulates self-splicing by promoting RNA assembly in vitro. AB - An Escherichia coli gene, stpA, has been identified and cloned based on its ability to suppress the Td- phenotype of a resident, splicing-defective phage T4 td (thymidylate synthase) gene. The stpA gene, which was localized to 60.24 min on the E. coli chromosome, encodes a 15.3-kDa protein. Overproduction of StpA in vivo led to an increase in td pre-mRNA levels and modest enhancement of td mRNA:pre-mRNA ratios. Consistent with its in vivo effect, purified StpA promoted RNA splicing in vitro, and facilitated RNA annealing and strand exchange with model substrates. These results suggest that StpA promotes splicing of the intron by binding RNA nonspecifically, resolving misfolded precursor molecules and facilitating association of critical base pair elements. Furthermore, proteinase K treatment of StpA-assembled precursors prior to the initiation of the splicing reaction still resulted in splicing enhancement, indicating that StpA is not required for the catalytic step, unlike the Neurospora splicing effector CYT-18, whose presence was necessary for catalysis to proceed. Together these results suggest that StpA has chaperone activity in vitro, with the property of promoting assembly of the precursors into an active conformation, in contrast to splicing effectors that stabilize the catalytically active intron structure. PMID- 7493325 TI - Titration of serine/arginine (SR) splicing factors during adenoviral infection modulates E1A pre-mRNA alternative splicing. AB - Alternative splicing of the adenovirus-2 E1A pre-mRNA involves the use of three 5' splice sites and is modulated during infection because the 13S mRNA and 9S mRNA reactions are predominant during the early and late periods, respectively. We had previously reproduced in vitro the 13S to 9S modulation with nuclear extracts isolated from infected HeLa cells and shown that high molecular weight viral RNAs are involved in this modulation, most likely by sequestering or titrating general splicing factors. To further test this hypothesis, we titrated splicing factors from an uninfected nuclear extract using competitor RNA or by progressive inactivation of splicing factors with monoclonal antibodies. We found that the 13S to 9S modulation occurs when titrating only with certain RNAs (essentially adenoviral RNAs), and also by progressively inactivating the 9G8 SR splicing factor. The demonstration that late nuclear extracts contain levels of active SR splicing factors limiting for the 13S reaction has been made by complementation experiments. We show that late nuclear extracts do not complement SR factor-deficient extracts, whereas late extracts treated with micrococcal nuclease complement them. Furthermore, complementation of late nuclear extracts with each of the three 30-35-kDa SR factors (9G8, SC35, and SF2/ASF) restores an efficient 13S mRNA reaction. Thus, our results provide evidence that the 13S to 9S modulation is triggered through a titration of SR factors required for the 13S mRNA reaction by major late transcripts that accumulate in nuclei late in infection. PMID- 7493326 TI - G.U base pairing motifs in ribosomal RNA. AB - An increasing number of recognition mechanisms in RNA are found to involve G.U base pairs. In order to detect new functional sites of this type, we exhaustively analyzed the sequence alignments and secondary structures of eubacterial and chloroplast 16S and 23S rRNA, seeking positions with high levels of G.U pairs. Approximately 120 such sites were identified and classified according to their secondary structure and sequence environment. Overall biases in the distribution of G.U pairs are consistent with previously proposed structural rules: the side of the wobble pair that is subject to a loss of stacking is preferentially exposed to a secondary structure loop, where stacking is not as essential as in helical regions. However, multiple sites violate these rules and display highly conserved G.U pairs in orientations that could cause severe stacking problems. In addition, three motifs displaying a conserved G.U pair in a specific sequence/structure environment occur at an unusually high frequency. These motifs, of which two had not been reported before, involve sequences 5'UG3' 3'GA5' and 5'UG3' 3'GU5', as well as G.U pairs flanked by a bulge loop 3' of U. The possible structures and functions of these recurrent motifs are discussed. PMID- 7493328 TI - A UV-induced, Mg(2+)-dependent crosslink traps an active form of domain 3 of a self-splicing group II intron. AB - In vitro irradiation of a15 gamma group II intron RNA with low doses of 254 nm UV light induces a single major crosslink. This crosslink was mapped within the domain 3 substructure of this RNA and one of the participating nucleotides was identified. When an RNA containing only the domain 3 substructure is irradiated under the same conditions, an intramolecular crosslink forms between two specific pyrimidines, one of them identical to the nucleotide crosslinked in the full length intron RNA. In both RNAs, the crosslink is magnesium ion-dependent and photoreversible. A trans assay for domain 3 function was developed and used to find that the crosslinked domain 3 RNA remains highly reactive. This suggests that crosslinking has trapped a functional, Mg(2+)-induced folded state of this group II intron substructure and that this folding is probably independent of the other domains of the intron. PMID- 7493327 TI - A short 5' splice site RNA oligo can participate in both steps of splicing in mammalian extracts. AB - A short 5' splice site RNA oligonucleotide (5'SS RNA oligo) undergoes both steps of splicing when a second RNA containing the 3' splice site region (3'SS RNA) is added in trans. This trans-splicing reaction displays the same 5' and 3' splice site sequence requirements as cis-splicing of full-length pre-mRNA. The analysis of RNA-snRNP complexes formed on each of the two splice site RNAs is consistent with the formation of partial complexes, which then associate to form the complete spliceosome. Specifically, U2 snRNP bound to the 3'SS RNA associates with U4/U5/U6 snRNP bound to the 5'SS RNA oligo. Thus, as expected, trans splicing depends on the integrity of U2, U4, and U6 snRNAs. However, unlike cis splicing, trans-splicing is enhanced when the 5' end of U1 snRNA is blocked or removed or when the U1 snRNP is depleted. Thus, the early regulatory requirement for U1 snRNP, which is essential in cis-splicing, is bypassed in this trans splicing system. This simplified trans-splicing reaction offers a unique model system in which to study the mechanistic details of pre-mRNA splicing. PMID- 7493329 TI - Structural and functional accommodation of nucleotide variations at a conserved tRNA tertiary base pair. AB - The U8:A14 tertiary base pair of transfer RNAs (tRNAs) stabilizes the sharp turn from the acceptor stem to the dihydrouridine stem. This tertiary base pair is important for the overall L-shaped tRNA structure. Inspection of tRNA sequences shows that U8:A14 is highly conserved. However, variations of U8:A14 are found in natural sequences. This raises the question of whether all 16 permutations of U8:A14 can be accommodated by a single tRNA sequence framework and by the bacterial translational apparatus. Here we expressed the wild type and 15 variants of U8:A14 of an alanine tRNA amber suppressor in Escherichia coli and tested the ability of each to suppress an amber mutation. We showed that 12 of the 15 variants are functional suppressors (sup+) and 3 are nonfunctional (sup-). Of the 12 functional suppressors, the G8:G14 variant is the most efficient suppressor, whose suppression efficiency is indistinguishable from that of the wild type. Analysis of tRNA structure with chemical probes and the lead-cleavage reaction, however, showed a distinct difference between the G8:G14 variant and the wild type. Thus, two different structures of E. coli tRNAAla/CUA share an identical functional phenotype in protein synthesis. The remaining 11 sup+ variants with reduced suppression efficiencies are likely to have other structural variations. We suggest that the variations of these sup+ mutants are structurally and functionally accommodated by the bacterial translational apparatus. In contrast, the three sup- mutants harbor variations that alter the backbone structure in the corner of the L. These variations are likely to reduce the stability of the tRNA inside the cell or, among others, to interfere with the ability of the tRNA to functionally interact with elongation factor Tu and with the ribosome. PMID- 7493330 TI - New proteins related to the Ser-Arg family of splicing factors. AB - A family of six highly conserved proteins that contain domains rich in alternating serine/arginine residues (SR proteins) function in the regulation of splice site selection and are required for splicing. Using a selective precipitation method, more than 35 proteins were detected in nuclear extracts of HeLa cells that co-fractionate with the defined SR family. Many of these proteins were recognized by three monoclonal antibodies that bind to distinct phosphoepitopes on SR proteins. Two of these SR-related proteins were identified as the nuclear matrix antigens B1C8 and B4A11, which previously have been implicated in splicing. A subset of SR proteins, in their phosphorylated state, are associated with spliceosome complexes through both steps of the splicing reaction, remaining preferentially bound to complexes containing the exon product. In contrast, other SR-related proteins appear to remain specifically associated with the intron-Iariat complex. The results indicate the existence of a potentially large group of SR-related proteins, and also suggest possible additional functions of SR proteins at a post-splicing level. PMID- 7493331 TI - Proceedings of the 5th Conference on Radioimmunodetection and Radioimmunotherapy of Cancer. Princeton, New Jersey, October 6-8, 1994. PMID- 7493332 TI - Labeling of polypeptides with technetium-99m using a dextran spacer. AB - Radioimmunoscintigraphy and other forms of scintigraphy are becoming increasingly important as tools in detection and evaluation of cancer diseases. Several radionuclides can be considered as radiolabel; however, 99mTc may be the most suitable because of easy availability and near ideal physical properties. This study describes how dextran is coupled to a monoclonal anticytokeratin antibody, epidermal growth factor, and somatostatin through stable amine bonds. The dextran moiety could then be labeled with 40-62 MBq of 99mTc. The labeling efficiency was 60-94%, determined by gel filtration or TLC. The specific binding could be considerably preserved after the dextranation, the antibody retained > 90% of its activity, epidermal growth factor bound with > 90% specificity to receptor expressing tumor cells, and somatostatin showed approximately 40% specific binding to rat adrenal homogenates. Because somatostatin only has a transient half-like in vivo, the in vivo plasma half-life of somatostatin-dextran was tested in normal female Sprague-Dawley rats. The result showed a strong enhancement of the plasma half-life that was dependent on two compartments, 4 and 8 h. PMID- 7493333 TI - Identification of metabolites of 111In-diethylenetriaminepentaacetic acid monoclonal antibodies and antibody fragments in vivo. AB - The in vivo fate of various 111In-labeled polypeptides has been the subject of many investigations. Intracellular metabolism has been studied through the use of 111In-labeled glycoproteins that are concentrated in the lysosome by receptor mediated endocytosis. These studies have indicated that the main lysosomal metabolite is 111In-chelate-epsilon-lysine, both in vitro and in vivo (Y. Arano et al., J. Nucl. Med., 35: 890-898, 1994; F. N. Franano et al., Nucl. Med. Biol., 21: 1023-1034, 1994). Since the vast majority of radiolabeled antibodies do not localize within the target tissue, an understanding of the metabolism of 111In labeled antibodies in nontarget tissues is important for the rational design of future radiolabeled antibodies. We investigated the in vivo metabolism of 111In DTPA3-conjugated antibody in female Sprague-Dawley rats using the anticolorectal carcinoma monoclonal antibody (MAb) 1A3 and MAb 1A3-F(ab')2. Livers and kidneys were harvested from rats injected with either intact MAb or MAb fragments and analyzed by gel filtration chromatography. Thirty-five % of the radioactivity from 111In-DTPA-1A3 MAb present in the liver was in the form of a low molecular weight species at 1 through 5 days. In contrast, 111In-DTPA-1A3-F(ab')2 was > 98% degraded to a low molecular weight species in the kidney after 1 day. In each case, the low molecular weight metabolites were collected and further analyzed by silica gel thin-layer chromatography, reversed phase high-performance liquid chromatography, and ion-exchange chromatography and compared to 111In-DTPA and 111In-DTPA-epsilon-lysine standards. In each system, the major metabolite co eluted with 111In-DTPA-epsilon-lysine, similar to the results obtained with 111In labeled glycoproteins that are delivered to lysosomes by receptor-mediated endocytosis. A minor metabolite that was more highly charged than 111In-DTPA was also observed. Analysis of urine and feces demonstrated that the main excretory product of both 111In-labeled intact 1A3 and 1A3-F(ab')2 was 111In-DTPA-epsilon lysine. Based on this data, we propose that 111In-DTPA-antibodies are degraded within lysosomes of nontarget organs such as the liver and kidneys. PMID- 7493334 TI - Site-specific modifications of light chain glycosylated antilymphoma (LL2) and anti-carcinoembryonic antigen (hImmu-14-N) antibody divalent f1agments. AB - Site-specific introduction of metal-chelating groups into F(ab')2 fragments of an antilymphoma antibody (LL2) possessing a natural Asn-linked light chain carbohydrate and an anti-carcinoembryonic antigen antibody (hImmu-14-N) grafted with a light chain carbohydrate site is described. For this purpose, four yttrium (and indium)-chelating agents were used, containing a primary amino group for antibody binding and 1-(4-substituted benzyl)diethylenetriaminepentaacetic acid as the metal-chelator, separated by structurally different additional linkers. Conjugates were prepared by reacting excess chelator with oxidized carbohydrate of F(ab')2 fragments, with or without a subsequent reduction step. The conjugates, with up to an average of 5.5 chelating groups attached to a F(ab')2 fragment, were readily labeled with 90Y and 111In and were found to retain antigen-binding ability in in vitro assays. Tumor targeting was demonstrated using a 88Y-labeled hImmu-14-N F(ab')2 carbohydrate-modified conjugate. 2 Pyridyldithiopropionic hydrazide was conjugated to the carbohydrate region, and the disulfide was selectively deprotected to the thiol group, which is reactive with reduced 99mTc. These initial experiments establish that light chain carbohydrate modification of F(ab')2 is as facile as with the Fc-region carbohydrate of intact IgG, and thereby offer the possibility of designing site specifically substituted F(ab')2 fragments with favorable pharmacokinetic properties. PMID- 7493336 TI - Experimental approaches to increase radiolabeled antibody localization in tumors. AB - Approaches have been developed to improve the localization of radiolabeled monoclonal antibodies (MAbs) in experimental tumors, to reduce their uptake in normal tissues, and, thus, to improve the time-dependent tumor: normal tissue (T:NT) ratios so that higher and more frequent doses of radionuclide could be used for radioimmunotherapy. These approaches involve three general strategies: (a) modifying antibodies or radiolabeling techniques; (b) increasing the clearance of radiolabeled MAbs; and (c) modifying tumor delivery, tumor antigen expression, or increasing tumor vascular permeability or blood flow. The use of animal models permits the assessment of a wide range of MAbs, radiolabeling conditions, and the efficacy of administration methods before their initial use in clinical trials. MAbs with specificity for binding to tumor-associated antigens or growth factor receptors expressed on tumor cells have been utilized in experimental studies of radiolabeled antibody targeting. Tumor-associated targets present on endothelial cells should be highly accessible to systemically administered radiolabeled MAbs. The use of indirect radio-iodination techniques and labile linker-chelates may provide an improvement in tumor retention and T:NT ratios. The addition or deletion of glycosylation to MAbs by alteration of recombinant immunoglobulin genes or by biochemical modification can alter the pharmacokinetics of blood and whole body clearance of radiolabeled MAbs. Genetically engineered chimeric or humanized MAbs have shown equivalent or greater tumor localization compared to murine MAbs. By using MAbs with greater affinity and avidity, an increase in the uptake and retention of radiolabeled MAbs in tumors and an increase in their therapeutic efficacy may be achieved. Several approaches in the administration methods of MAbs have been developed in an attempt to improve tumor localization and therapeutic results and to reduce toxicity. These approaches include: (a) predosing with unlabeled antibody before administering a radiolabeled MAb; (b) using a mixture or "cocktail" of MAbs rather than a single radiolabeled antibody; and (c) administering multiple doses of radiolabeled MAbs. Various approaches have been tested for increasing the blood clearance of radiolabeled MAbs and, thus, for increasing the T:NT ratio. It has been found that compared to intact antibody, the smaller antibody fragments (F(ab')2, Fab, or single-chain Fv) can bind to tumor cells with a more homogeneous distribution. The antibody fragments and domain deletions often have a more rapid catabolism in blood, in tumors, and in normal tissues than an intact antibody does. In general, the use of antibody fragments leads to higher T:NT ratios but a lower percentage of injected dose delivered to the tumor. PMID- 7493335 TI - Prelabeling of chimeric monoclonal antibody L6 with 90yttrium- and 111indium 1,4,7,10-tetraazacyclododecane-N,N',N",N"'-tetraacetic acid (DOTA) chelates for radioimmunodiagnosis and therapy. AB - 90Y and 111In have been attached to chimeric monoclonal antibody L6 with a bifunctional chelating agent (DOTA-peptide isothiocyanate). The bifunctional chelating agent was prelabeled with either radiometal and then conjugated to the antibody. Studies in human patients showed excellent 111In single-photon emission computed tomography images of breast cancer lesions 24 h after injection. PMID- 7493337 TI - On the validity of "functional affinity" determination for antibodies binding to cell surface antigens or other polyvalent antigens. AB - A widely used method of comparing different antibodies (Abs) is to determine their "functional affinities." This value is supposedly a constant that reflects the basic binding interaction between antibody and antigen and, if it is determined under standard conditions, allows the comparison of Abs used by different laboratories. However, I present here both theoretical and experimental evidence that, for Abs binding bivalently, functional affinity determinations seem to be invalid. Experimental data were obtained with erythrocyte targets, to eliminate interference due to the internalization or catabolism of bound Ab, but similar results have been obtained with tumor target cells. Several fundamental discrepancies between the theoretical expectations and the actual values obtained were demonstrated, all of which can be attributed to the effects of bivalent binding. One discrepancy is that the functional affinity determined did not equal the ratio of the rate constants for association and dissociation. A second is that the functional affinity was not a constant but, rather, depended on trivial experimental conditions, such as the volume of incubation. In addition, it has been recognized previously that functional affinity is affected strongly by the structure of a multivalent antigen and, in particular, by any change that makes it more or less likely that an Ab will bind bivalently. The value of other methods of comparing the avidity of different Abs is discussed. In many situations, Ab binding can be considered predominantly irreversible for practical purposes. PMID- 7493338 TI - Preclinical analysis of intraperitoneal administration of 111In-labeled human tumor reactive monoclonal IgM AC6C3-2B12. AB - An IgM lambda human tumor cell-reactive monoclonal antibody was developed that reacts with cells of ovarian cancer, colorectal cancer, breast cancer, and certain other malignancies. The monoclonal antibody AC6C3-2B12, which was obtained from a recent recloning, was purified from tissue culture supernatants and analyzed by high-performance liquid chromatography and sodium dodecyl sulfate PAGE. An animal model was developed in which human tumors grew either as solid peritoneal metastases or as s.c. nodules utilizing the human colorectal carcinoma cell line SW620. The biodistribution of 111In-labeled IgM conjugate was studied after i.v. or i.p. administration in nude mice bearing an s.c. xenograft or peritoneal tumor lumps of a human colorectal carcinoma (SW620). IgM administered i.v. cleared rapidly from blood and was deposited mainly in the liver [50% injected dose/g (ID)/g)], pancreas (20% ID/g), and kidney (10% ID/g) at 24 h. Tumor deposition was low (< or = 1.0% ID/g) in the s.c. tumor xenograft. In contrast, high tumor targeting (29% ID/g) was found in peritoneal tumor lumps after i.p. administration of 111In-labeled IgM. The biological half-life of IgM in the tumor was 100 h. Long peritoneal residence time (t 1/2 = 67 h) and low liver uptake (7% ID/g) were observed after i.p. administration. Blood activity was < 1% of the injected activity. Tumor:normal organ ratios were high (range, 2 290) from 2 to 144 h after i.p. administration. Whole body autoradiograms at 24 h after i.p. 111In-labeled IgM administration confirmed the biodistribution results. In normal beagle dogs, 75% of the i.p.-administered 111In-IgM decayed in the peritoneal cavity. The majority of the remaining radioactivity was taken up by mediastinal lymph nodes. Biological half-life in both locations was approximately 137 h. The i.p. administration of intact, specific radiolabeled IgM provides prolonged retention of radioactivity in tumor, low normal tissue uptake, a long peritoneal residence time, and very limited spillover of IgM into the circulation. This approach offers a promising new method for the diagnosis and treatment of certain patients with peritoneal carcinomatosis. PMID- 7493339 TI - Initial studies of monoclonal antibody PAM4 targeting to xenografted orthotopic pancreatic cancer. AB - To resemble the clinical presentation of pancreatic cancer in an animal model more closely, we developed an orthotopic xenograft of CaPan-1 human pancreatic cancer in athymic nude mice. Within 3 weeks after implantation into the body and head of the pancreas, animals had palpable tumors. By 8 weeks, metastases to the liver and spleen were observed, and at 10-14 weeks, ascites formation, with and without seeding of the diaphragm, and jaundice were evident. Thus, this tumor model exhibited many of the most common features of human pancreatic cancer. Radiolabeled monoclonal antibody PAM4 showed specific localization of the primary orthotopic and metastatic tumors. On day 3, PAM4 accumulation within the primary tumor (0.5 g) was 11.3 +/- 5.1% injected dose/g with a localization index of 11.3 +/- 4.0. The estimated tumor:blood radiation dose ratio for PAM4 was 4:1, whereas a nonspecific antibody (Ag8) would provide only 40% of the blood dose to the tumor. Based on these observations, animals bearing 4-week-old orthotopic tumors (estimated volume, 0.25 cm3) were administered either 131I-labeled PAM4, 350 microCi, or nonspecific Ag8, 350 microCi, and compared with an untreated control group. Radiolabeled PAM4 provided a significant (P < 0.001) increase in survival time with less morbidity compared with the untreated control group, whereas nonspecific Ag8 was not significantly different from the control group. These studies provide a rationale for initiating a Phase I clinical study for detection and therapy of pancreatic cancer with PAM4. PMID- 7493341 TI - Improving the balance between treatment and diagnosis: a role for radioimmunodetection. AB - Despite major advances in diagnostic testing, including the introduction and widespread availability of high-resolution computed tomography (CT) and magnetic resonance imaging, inadequate diagnostic information still interferes with proper management of many patients with cancers. This is particularly true for recurrent colorectal cancer, for example. In the course of this symposium, significant advances have been reported which are likely to improve management of this clinical situation. 111In, oncoscint for colorectal and ovarian cancer imaging, has been approved for single use only, and is a product licensed by the Food and Drug Administration. It has been shown to be significantly more effective than CT for detecting the presence of disease that is confined to the abdomen outside the liver. This agent is very useful in a limited role. A larger opportunity awaits other preparations reported at this conference, especially 99mTc-labeled Immu-4 carcinoembryonic antigen, which is significantly better than CT for determining resectability of recurrent cancer (T. Behr et al., Cancer Res. 55 (Suppl.): 5777S 5785S, 1995). The 99mTc-labeled compound preparations offer the advantages of low immunogenicity, excellent imaging energies of 99mTc, and "same-day" imaging. Even the most effective cancer treatment such as surgical resection, if applied to a patient who basically does not need it, can be a needless expense and a trauma to the patient. To date, our emphasis in oncology research has been heavily weighted toward developing new therapies. The success of radioimmunodetection is one indication of why it is time for a paradigm shift, during which we can move toward a more balanced program that emphasizes both diagnosis and therapy. To achieve this we must urge research institutions such as the National Cancer Institute and American Cancer Society to make major investments in the diagnostic aspects of cancer care. With the knowledge base that we have now, we can make improvements in patient care by emphasizing development of improved diagnostic methods and support for cost-effectiveness studies for developed methods, in order that currently available treatments can be more intelligently applied. PMID- 7493342 TI - Fusion of immunoscintigraphy single photon emission computed tomography (SPECT) with CT of the chest in patients with non-small cell lung cancer. AB - In non-small cell lung cancer (NSCLC), accurate staging is critical in deciding between potentially curative surgery and palliative treatment. Image registration, or fusion, combines the unique functional information provided by SPECT imaging with the excellent anatomic detail offered by computed tomography (CT) or magnetic resonance imaging to better characterize the information provided by each separate modality. In this study, we explored the role of fusion of immunoscintigraphy SPECT with CT in the staging of NSCLC. We fused chest CT with 99mTc-labeled IMMU-4 anti-carcinoembryonic antigen Fab' antibody fragment SPECT in 14 patients with NSCLC using a landmark-based algorithm. The algorithm's accuracy was a measure from the center-to-center distance and the percentage overlap of two regions of interest: one drawn on CT and warped onto SPECT, the other drawn directly on the SPECT. We found that the average center-to-center distance was 1.3 +/- 0.8 pixels. Average overlap was 46 +/- 20%. CT-SPECT fusion helped differentiate tumor from normal blood pool, necrotic areas within viable tumor, tumor recurrence from scar, and malignant lymphadenopathy from hyperplasia. We conclude that fusion of CT and SPECT augments the information provided by each separate modality. Future clinical applications of fusion in NSCLC staging using immunoscintigraphy appear promising. PMID- 7493340 TI - A monoclonal antibody-cobra venom factor conjugate increases the tumor-specific uptake of a 99mTc-labeled anti-carcinoembryonic antigen antibody by a two-step approach. AB - Monoclonal antibodies (mabs) have considerable potential for specific cancer treatment. However, due to the antigen heterogeneity and especially the low uptake in solid tumors, mabs have not been used successfully in most clinical trials to date. This study investigates the effects of a mab-cobra venom factor (CVF) conjugate in vitro and in vivo in an orthotopic pancreatic cancer model using nude rats. CVF, a nontoxic glycoprotein from cobra venom, permanently activates the alternative pathway of complement. Coupled to a mab with tumor binding properties, the complement activation can be targeted to the tumor tissue. We studied the activity of a mab CA19-9-CVF conjugate with the human pancreatic cancer cells PancTu I. PancTu I cells express the complement resistance factors CD46, CD55, and CD59, as we demonstrated by immunostaining, an observation that may explain the lack of cytotoxicity of the CA19-9-CVF conjugate. However, using ELISA, Western blot, and immunostaining, we showed that CA19-9-CVF activates the complement cascade, including the release of the anaphylatoxin C3a, a mediator of an inflammatory reaction. The in vivo studies of CA19-9-CVF-treated nude rats showed an increased tumor infiltration by natural killer cells and macrophages. The tumor uptake of 99Tc-labeled anti carcinoembryonic antigen antibody was increased approximately 2-fold in rats pretreated with 70 micrograms of CA19-9-CVF, compared to animals that received an equimolar mixture of noncoupled mab and CVF. This study indicates the value of mab-CVF conjugates in adjuvant immunotherapy. mab-CVF conjugates might be useful in pretargeting approaches by increasing the uptake of a therapeutic mab. PMID- 7493343 TI - Improved staging of B-cell non-Hodgkin's lymphoma patients with 99mTc-labeled LL2 monoclonal antibody fragment. AB - Radioimmunodetection (RAID) with the 99mTc-labeled Fab' fragment of monoclonal antibody (MoAb) LL2 has been reported to have a high lesion detection rate for malignant lymph nodes as well as for visceral and skeletal tumor masses (20). Our purpose in this study was to evaluate the safety and staging efficacy of the 99mTc-labeled Fab' fragment of MoAb LL2 in patients with various grades and stages of B-cell non-Hodgkin's lymphoma (NHL). Thirty adult patients, 13 male and 17 female, ranging in age from 20 to 80 years, with at least one biopsy-proved malignant node (> or = 0.5 cm) and a Karnofsky performance score of > or = 60% were enrolled in this study. Patients underwent selected planar and single photon emission computed tomographic imaging at 6 and 18 h after receiving an i.v. infusion of 0.25-1 mg of LL2 Fab' fragment labeled with 25-30 mCi of 99mTc. RAID findings were compared with physical examination, chest radiography, computed tomography, magnetic resonance imaging, and bone and 67Ga scan findings. The RAID scan was positive in all but three patients. The sensitivity for known lesions was 90% and for suspected lesions, 89%, with an overall positive predictive value of 96%. Twenty-nine of the 30 patients had either low- or intermediate-grade NHL. Fifteen of 16 (94%) low-grade patients were correctly staged by RAID; three of these patients were correctly upstaged. Twelve of the 13 (92%) intermediate-grade patients were correctly staged by RAID; two of these patients were correctly upstaged. The high-grade NHL patient was staged correctly by RAID. Infused doses of 99mTc-labeled LL2 Fab' of 0.5 and 1.0 mg did not affect lesion sensitivity. The RAID sensitivity decreased as the total tumor burden increased > or = 100 g. On the basis of these initial results, the 99mTc-labeled Fab' fragment of MoAb LL2 (LymphoScan) seems to yield useful clinical information, especially for the staging of B-cell NHL patients who do not have bulky disease (i.e., tumor burdens of < or = 100 g). PMID- 7493344 TI - 67Ga citrate versus 99mTc-labeled LL2-Fab' (anti-CD22) fragments in the staging of B-cell non-Hodgkin's lymphoma. AB - The value of 67Ga citrate scanning as a transferrin receptor agent was compared in this study with a 99mTc-labeled anti-CD22 (B-cell) Fab' fragment (LL2) in patients with low- and high-grade B-cell non-Hodgkin's lymphoma (NHL). Thirteen patients with histologically confirmed NHL were examined prospectively with both radiopharmaceuticals within 8 days. The results of immunoscintigraphy were compared with those of 67Ga scanning and the clinical and radiological workup (computed tomography, ultrasound, and magnetic resonance imaging) of the patients. The overall sensitivity of 67Ga citrate and 99mTc-labeled LL2 fragment was each 80% in a total of 43 lesions. Low-grade lymphoma patients had a higher sensitivity in LL2 imaging (82% versus 71%), and high-grade lymphoma patients in 67Ga citrate scanning (100% versus 75%). The target:background ratio in low-grade NHL for LL2 was 1.43 +/- 0.3:1 versus 1.8 +/- 0.5:1 in 67Ga scans; in high-grade NHL, 1.49 +/- 0.35:1 versus 2.2 +/- 0.8:1, respectively. Single-photon emission computed tomography imaging was necessary in 21.7% of the patients 4 h after injection to localize the lesions. In conclusion, the overall sensitivity of 99mTc-labeled LL2 is comparable to 67Ga citrate scanning in patients with B-cell NHL. 99mTc-labeled LL2 antibodies are rapid to use, are safe, and need a shorter imaging time (24 h versus 72 h). Because of these advantages, 99mTc-labeled LL2 may be superior to 67Ga scanning for the staging of lymphoma patients. PMID- 7493345 TI - Radioimmunoscintigraphy of colorectal carcinoma using technetium-99m-labeled, totally human monoclonal antibody 88BV59H21-2. AB - Radioimmunoscintigraphy (RIS) using human monoclonal antibodies offers the important clinical advantage of repeated imaging over murine monoclonal antibodies by eliminating the cross-species antibody response. This article reports a Phase I-II clinical trial with Tc-99m-labeled, totally human monoclonal antibody 88BV59H21-2 in patients with colorectal carcinoma. The study population consisted of 34 patients with colorectal cancer (20 men and 14 women; age range, 44-81 years). Patients were administered 5-10 mg antibody labeled with 21-41 mCi Tc-99m by the i.v. route and imaged at 3-10 and 16-24 h after infusion using planar and single-photon emission computed tomographic (CT) techniques. Pathological confirmation was obtained in 25 patients who underwent surgery. Human antihuman antibody (HAHA) titers were checked prior to and 1 and 3 months after the infusion. RIS with Tc-99m-labeled 88BV59H21-2 revealed a better detection rate in the abdomen-pelvis region compared with axial CT. The combined use of both modalities increased the sensitivity in both the liver and abdomen pelvis regions. Ten patients developed mild adverse reactions (chills and fever). No HAHA response was detected in this series. Tc-99m-labeled human monoclonal antibody 88BV59H21-2 RIS shows promise as a useful diagnostic modality in patients with colorectal cancer. RIS alone or in combination with CT is more sensitive than CT in detecting tumor within the abdomen and pelvis. Repeated RIS studies may be possible, due to the lack of a HAHA response. PMID- 7493346 TI - Targeting of liver metastases of colorectal cancer with IgG, F(ab')2, and Fab' anti-carcinoembryonic antigen antibodies labeled with 99mTc: the role of metabolism and kinetics. AB - The aim of this study was to investigate targeting of the liver metastases by directly 99mTc-labeled complete (IgG) and fragmented antibodies [F(ab')2 and Fab'] in relation to their kinetics and metabolic fate. A total of 127 patients with metastatic colorectal cancer were examined [IgG1, BW 431/26 (Behringwerke, Marburg, Germany) n = 50; F(ab')2, F023C5 (Sorin Biomedica, Saluggia, Italy) n = 58; Fab', IMMU-4 (Immunomedics, Morris Plains, NJ) n = 19]. Native monoclonal antibodies (MAbs), serum samples from 10 min to 24 h postinjection (p.i.), and urine were analyzed by gel filtration chromatography. Kinetic data were deduced from whole-body and single-photon emission computed tomographic scans, performed 10 min to 24 h p.i. (region-of-interest technique). In BW 431/26, 96% of injected activity was labeled IgG1; in F023C5, 29% was F(ab')2, and 71% was Fab'; and in IMMU-4, 92% was Fab', and 8% was F(ab')2. Serum half-lives were: IgG1, 36 h (liver uptake predominant); F(ab')2, 16 h; and Fab', 4 h (renal uptake predominant). All MAbs were metabolized, fragments more rapidly than IgG, to low molecular-weight products and excreted into the urine (e.g., Tc-cystine). In targeting liver metastases, sensitivities were found to be higher for fragments (44.1, 72.5, and 80% for BW 431/26, F023C5, and IMMU-4, respectively) but at significantly lower tumor:background ratios than with IgG (1.78 +/- 0.29 versus 1.29 +/- 0.11 and 1.43 +/- 0.53; P < 0.01). With IgG, there was a continuous tumor uptake over 24 h, whereas with fragments, the maximal uptake occurred mostly within 1 h, with subsequent clearance being slower for antigen-bound activity than for nonspecific background. Hence, diagnosis was possible mostly after 4 h with fragments but often not before 24 h with IgG. These results show that the higher sensitivity of fragments in liver lesion targeting at earlier p.i. times does not rely on an increased antibody uptake but on a more rapid clearance of nonspecific background activity due to faster metabolism and excretion. Intact MAbs show a slow, continuous uptake, leading to higher tumor:background ratios at later p.i. times, often beyond the imaging possibilities of 99mTc. PMID- 7493347 TI - Diagnostic accuracy and tumor-targeting kinetics of complete versus fragmented 99mTc-labeled anti-carcinoembryonic antigen antibodies: an intraindividual comparison. AB - The goal of this study was to intraindividually compare a complete versus a fragmented, directly 99mTc-labeled, monoclonal anti-carcinoembryonic antigen (CEA) antibody, with respect to their antigen-targeting kinetics, sensitivity, and diagnostic accuracy in patients with CEA-expressing tumors. Twenty-five patients were investigated with the 99mTc-labeled anti-CEA IgG1 BW 431/26 and the F(ab')2/Fab' fragment mixture F023C5 within 7 days. For quantitative analysis, the region of interest technique was applied to planar scans, whole-body scans, and single photon emission computed tomography slices 10 min to 48 h postinjection (PI). Final correlations were performed according to the histopathology after surgery or biopsy. Earliest tumor detection was possible with complete IgG1 4 h PI (52% of finally positive lesions). Twenty-four- or even 48-h scans were necessary in 48% of finally positive lesions; tumor detection with fragments was possible in 17% at 1 h PI and in 94% at 4 h PI. With both monoclonal antibodies, in 35%, single photon emission computed tomography was necessary for tumor detection. Absolute antibody uptake in tumor lesions was higher with complete monoclonal antibodies than with fragments. The sensitivity of fragments was higher in detecting primary tumors, local recurrences, and lymph node metastases. For detection of liver metastases, sensitivity was also higher for fragments than for IgG (87 versus 73%), but in scintigraphically positive lesions, tumor:background ratios were significantly lower with fragments (1.26 +/ 0.12 versus 1.70 +/- 0.32; P < 0.01). Therefore, fragments seem to be more suitable for earlier detection of lesions known for their good vascularization, vascular permeability, and antigen accessibility, such as local recurrences, lymph node metastases, and peritoneal carcinomatoses. In liver metastases (high interstitial pressure, low vascular leakage), sensitivity of fragments is higher, but their rapid serum and whole-body clearance lead to a lower absolute antibody uptake, with the consequence of significantly lower tumor:background ratios than with IgG. PMID- 7493348 TI - Human biodistribution of [111In]diethylenetriaminepentaacetic acid-(DTPA)-D [Phe1]-octreotide and peroperative detection of endocrine tumors. AB - Requisites for preoperative and intraoperative tumor localization with [111In]diethylenetriaminepentaacetic acid-D-[Phe1]-octreotide scanning were explored in 23 patients with endocrine tumors (15 carcinoids, 4 insulinomas, and single cases of gastrinoma, medullary thyroid carcinoma, aldosteronoma, and paraganglioma). The patients were subjected to Octreoscan single photon emission computed tomographic examination prior to surgery and well counter investigation of nuclide uptake in tumors and normal tissues sampled at surgery. Somatostatin receptor-positive tumors demonstrated efficient nuclide accumulation with mean tumor:blood radioactivity ratios of 180-370 (for carcinoids and insulinoma), compared with tissue:blood ratios of 302 for spleen, 42 for liver, and < 10-15 in other normal tissues (pancreas, small intestine, and mesenteric fat). Inefficient preoperative visualization of lesions was related to inconspicuous size, as for primary intestinal carcinoids, tiny liver metastases, and a single small insulinoma. High background activity, pronounced tumor fibrosis, and meager accumulation of tracer also interfered with visualization. Tumor deposits in organs with low background activity (such as carcinoid mesenteric metastases and endocrine pancreatic tumors) were generally most readily detected. Intraoperative investigations with hand-held gamma detector probes were disturbed by obvious high background activity. These investigations revealed two preoperatively unrecognized primary intestinal carcinoids, which, however, were both palpable during surgery. These studies, therefore, had little impact on the surgical strategy. PMID- 7493349 TI - Somatostatin receptor scintigraphy in patients with carcinoid tumors: comparison between radioligand uptake and tumor markers. AB - We have performed 100 scintigraphic investigations using [111In diethylenetriaminepentaacetic acid-D-Phe1]octreotide (111In-octreotide) single photon emission tomography (SPECT) in patients with carcinoid tumors. One or several lesions could be detected in 77 cases, and true negative results were obtained in 11 cases. There were false-negative results in 12 cases compared with results from conventional radiological methods. The ratio between the SPECT signals from the area with the highest uptake and normal lung was used as a tumor:background ratio. An attenuation correction was made in all investigations. We found that lesions in untreated patients had lower tumor:background ratios compared with those in patients treated with somatostatin analogues (medians, 10 and 40, respectively; P < 0.001) or IFN (median, 23; P = 0.03). In untreated patients, there was a correlation between the tumor:background ratio and the levels of urinary 5-hydroxyindoleacetic acid (U-5HIAA) and p-chromogranin A. The data obtained in the present investigation indicate that somatostatin receptor expression might be influenced by the treatment; i.e., a higher tumor:background ratio is found in patients treated with either somatostatin analogues or IFN. Furthermore, it was found that somatostatin receptor expression correlates with the levels of U-5HIAA and p-chromogranin A in untreated patients, and that 111In octreotide SPECT scintigraphy is more likely positive in patients with elevated U 5HIAA values. This indicates that somatostatin receptor expression and elevated U 5HIAA are more likely present in patients with highly differentiated tumors and, thus, could be of prognostic value. PMID- 7493351 TI - Reconciliation of tumor dose response to external beam radiotherapy versus radioimmunotherapy with 131iodine-labeled antibody for a colon cancer model. AB - Reported doses of external beam radiotherapy and radioimmunotherapy (RIT) to produce equivalent therapeutic effects are inconsistent, with many proposed causes. Calculations of effective dose were performed for the case of LS174T human colon cancer xenografts, where a 60Co single fraction exposure (6 Gy) was matched with 131I-labeled 17-1A monoclonal antibody therapy (300 microCi injection, 19 +/- 2 Gy using the Medical Internal Radiation Dose uniform isotropic model). Measured three-dimensional dose-rate distributions were used to form a time-dependent description of the dose-rate nonuniformity. Included in the calculation of RIT effective dose was energy loss, dose nonuniformity, dose-rate dependence, hypoxic fraction, and cell proliferation. The calculations assumed the linear quadratic model for cell survival with alpha = 0.3 Gy-1, alpha/beta = 15 to 25 Gy, and mu = 0.46 h-1. The biologically effective dose for the single fraction 60Co exposure was 7.4 to 8.4 Gy. Estimates of dose efficiency factors consecutively applied to the RIT dose estimate were: (a) energy loss external to the tumor (x0.85); (b) effect of dose nonuniformity on cell survival (x0.65); and (c) effect of correlation of dose nonuniformity with cell proliferation rate (x1.08). The resulting effective dose for RIT was 11.4 Gy for tumor regrowth. This analysis substantially reconciles external beam radiotherapy/RIT dose response results for this tumor model to within experimental uncertainties. PMID- 7493350 TI - Metastatic hormone-refractory prostatic adenocarcinoma expresses somatostatin receptors and is visualized in vivo by [111In]-labeled DTPA-D-[Phe1]-octreotide scintigraphy. AB - Thirty-one patients with metastatic hormone-refractory prostatic adenocarcinoma were investigated scintigraphically with the 111In-labeled somatostatin analogue [DTPA-D-Phe1]-octreotide (OctreoScan) and with 99mTc-labeled HDP. In vitro somatostatin receptor autoradiography was performed on biopsies obtained from eight patients with hormone-refractory prostatic adenocarcinoma. In 30 of 31 patients (94%), at least one metastasis was positive at OctreoScan scintigraphy. Of the 346 lesions detected with 99mTc-labeled HDP bone scintigraphy, 128 were visualized with the OctreoScan technique, thus accounting for a 37% detection rate. Two uptakes on OctreoScan could not be identified on bone scintigraphy and were, thus, assessed as false positive. The biopsies of the eight patients disclosed a low density of receptors, localized on the tumor cells, as demonstrated with receptor autoradiography. Two patients with untreated metastatic prostatic adenocarcinoma were investigated in vivo before the start of endocrine therapy. However, none of the lesions detected by bone scintigraphy in these patients could be visualized with the OctreoScan technique. Positron emission tomography using [11C] methionine showed a decreased uptake in a metastatic index lesion in a patient treated with octreotide. It is concluded that hormone-refractory prostatic adenocarcinoma expresses somatostain receptors both in vitro and in vivo. The results obtained form the basis for the development of a new tool for in vivo characterization and of a new treatment strategy in patients with hormone-refractory prostatic adenocarcinoma. PMID- 7493352 TI - Radiation-absorbed dose estimates to normal organs following intraperitoneal 186Re-labeled monoclonal antibody: methods and results. AB - The radiation-absorbed dose was estimated following i.p. administration of a 186Re-labeled murine antibody, NR-LU-10, in 27 patients with advanced ovarian cancer. Data for the dosimetry estimation were obtained from quantitative gamma camera imaging and gamma counting of serum and i.p. fluid radioactivity. A peritoneal cavity model was used to estimate the dose to normal organs from radioactivity within the peritoneal cavity. Estimates of radiation-absorbed dose to normal organs in rad/mCi administered (mean + SD) were: whole body, 0.7 + 0.3; marrow, 0.4 + 0.1; liver, 1.9 + 0.9; kidneys, 0.2 + 0.2, and intestine, 0.2 + 0.2. The radiation-absorbed dose estimates to the normal peritoneal surface varied depending on the volume of fluid infused and whether the activity was measured by the gamma camera or from the peritoneal fluid samples. Using gamma camera data, the peritoneal surface dose ranged from 7 to 36 rads/mCi; when using the peritoneal fluid sample data, the dose ranged from 2 to 25 rads/mCi. Myelosuppression, observed in several patients, correlated best with marrow dose estimates based on the serum radioactivity, and significant toxicity was observed at marrow doses greater than 100 rads. The noninvasive methods of dose estimation for i.p. administration of radioimmunoconjugates provided reasonable absorbed dose estimates when compared with previously described, more invasive methods. PMID- 7493353 TI - Threshold estimation in single photon emission computed tomography and planar imaging for clinical radioimmunotherapy. AB - Thresholding is the most widely used organ or tumor segmentation technique used in single photon emission computed tomography (SPECT) and planar imaging for monoclonal antibodies. Selecting the optimal threshold requires a priori knowledge (volumes from CT or magnetic resonance) for the size and contrast level of the organ in question. Failure to select an optimal threshold leads to overestimation or underestimation of the volume and, subsequently, the organ absorbed dose value in radio-immunotherapy. To investigate this threshold selection problem, we performed a phantom experiment using six lucite spheres ranging from 1 to 117 ml and filled with a uniform activity of 1 microCi/ml Tc 99m. These spheres were placed at the center and off-center locations of a Jasczsak phantom and scanned with a three-headed gamma camera in SPECT and planar modes. Target-nontarget (T:NT) ratios were changed by adding the appropriate activity to the background. A threshold search algorithm with an interpolative background correction was applied to sphere images. This algorithm selects a threshold that minimizes the difference between the true and measured volumes (SPECT) or areas (planar). It was found that for spheres equal to or larger than 20 ml [diameter (D) > 38 mm] and T:NT ratios higher than 5:1, mean thresholds at 42% for SPECT and 38% for planar imaging yielded minimum image segmentation errors, which is in agreement with current literature. However, for small T:NT ratios (< 5:1), the threshold values as high as 71% for SPECT and 85% for planar imaging were substantially different than those fixed thresholds for large spheres (D > 38 mm). Hence, the use of fixed thresholds in low contrasts and with tumor and organ sizes of clinical interest (25 < or = D < or = 50 mm) may result in limited volume estimation accuracy. Therefore, we have provided the investigator a method to obtain the threshold values in which the proper threshold can be selected based on the organ and tumor size and image contrast. By measuring and calibrating the proper threshold value derived through machine specific phantom measurements, a more accurate volume and activity quantitation can be performed. This, in turn, will provide tumor-absorbed dose optimization and greater accuracy in the measurement of potentially subacute, toxic absorbed doses to normal organs for patients undergoing radioimmunotherapy. PMID- 7493354 TI - Estimates of red marrow dose by sacral scintigraphy in radioimmunotherapy patients having non-Hodgkin's lymphoma and diffuse bone marrow uptake. AB - According to the recommendations of the Dosimetry Task Group of the American Association of Physicists in Medicine, blood-derived estimates of the red marrow (RM) dose from radiolabeled monoclonal antibodies (MAbs) are valid only if the RM is devoid of any specific uptake. There is, therefore, a clear need for an alternative method for estimating the RM dose in patients receiving MAbs that target normal or abnormal (malignant) bone marrow elements. Radiolabeled LL2, an anti-B-cell murine MAb, targets normal B cells and malignant lymphoma cells in the RM. This may result in an increased radiation dose to the RM through neighboring targeted activity. We investigated whether imaging-based estimates of the RM dose, particularly using sacral scintigraphy, correlate with myelotoxicity in non-Hodgkin's lymphoma patients who received 131I-LL2. The sacrum-based RM dose (RMs) was estimated from sacral activity by assuming that 9.9% of the total adult RM is contained in the sacrum. The sacrum was not used if there was focally increased or decreased sacral uptake. Myelotoxicity was assessed based on Radiation Therapy Oncology Group criteria. Twelve of 21 non-Hodgkin's lymphoma patients treated had adequate imaging, dosimetry, and follow-up to evaluate myelotoxicity. Eight of these patients had diffusely increased RM uptake on their MAb scans. The average estimated RMs in the eight patients was 168 +/- 62 cGy (mean +/- SD) with only 50 mCi 131I-LL2. Six of these patients (75%) developed grade 3 or 4 myelotoxicity. In contrast, the average RMs in the four patients who did not have any enhanced uptake on their scans was 71 +/- 30 cGy (P < 0.02). None of these patients developed grade 3 or 4 toxicity. These results suggest that image-based estimates of the RM dose may be predictive of myelotoxicity and should be used in patients with diffuse RM uptake on their scans. PMID- 7493355 TI - Overview of studies on experimental radioimmunotherapy. AB - Experimental radioimmunotherapy (RIT) has significantly contributed to the development of clinical RIT. In this overview, the current status of experimental RIT is reviewed, including general principles of RIT and determinants of RIT effects. Areas of active research are reviewed, and the usefulness of multicell spheroids is compared to animal models. The radiobiology of RIT is discussed, and studies that have compared the relative efficacy of RIT with external beam radiation therapy are summarized. Approaches for increasing the therapeutic index of RIT are reviewed, including improvements in antibodies, labeling/chelation chemistry, selection of radionuclides, delivery, fractionated therapy, clearance of unbound radiolabeled monoclonal antibodies, protection of normal tissues, radiosensitization of tumors, utilization of colony-stimulating factors and bone marrow transplantation, and the use of novel targets for RIT and topoisomerase I inhibitors. RIT is a promising new therapy for a wide variety of malignancies that can best be optimized by continued research in the field of experimental RIT. Important areas of future research are discussed that may ultimately potentiate the efficacy and decrease the toxicity of RIT and help determine how to optimally combine RIT with other therapeutic modalities. PMID- 7493356 TI - Yttrium-90 chimeric L6 therapy of human breast cancer in nude mice and apoptosis related messenger RNA expression. AB - Radioimmunotherapy (RIT) in breast cancer patients using I-131-chimeric L6 (ChL6) and in human breast cancer xenografts in nude mice using Y-90-1,4,7,10 tetraazacylododecant N,N',N",N"'-tetraacetic acid-peptide ChL6 (Y-90-ChL6) has shown promise. Tumor cell response to low-dose rate (5-25 rads/h) irradiation from Y-90-ChL6 RIT, therefore, was correlated with levels of tumor cell mRNA for selected genes linked to programmed cell death (apoptosis). Three groups of 10-16 mice with 1-2 HBT 3477 xenograft tumors were treated with 100, 150, or 250 microCi Y-90-ChL6. Three tumors were taken before and two tumors each were taken 3, 6, and 24 h after injection of 150 microCi Y-90-ChL6. Tumor expression of mRNA was amplified by PCR for p53, PIC1, c-myc, and transforming growth factor-beta 1; quantitated; and standardized to N-ras. Tumors received radiation doses of 2000, 3000, and 5000 rads, respectively, for the groups of mice that received 100, 150, and 250 microCi Y-90-ChL6, and tumor regression occurred in each group, with mean tumor volumes decreased by 10, 50, and 95% at nadir after Y-90-ChL6 injection. At the highest dose level, 30% of mice had complete remissions, and no treatment deaths occurred, although tumors subsequently recurred. Continuous up-regulation of transforming growth factor-beta 1 and c-myc mRNA expression was observed from 3 to 24 h after treatment. Expression of p53 and PIC1 increased at 3 h and subsequently decreased to the untreated control levels. These observations are consistent with previous observations of early responses of p53 and PIC1 to cellular DNA damage and subsequent G1 cell cycle arrest or apoptosis. Apoptosis associated gene expression patterns observed in this tumor model provide evidence that changes are initiated in the first 24 h of RIT associated with radiation doses of 100-700 rads. These preliminary data suggest that insight into the molecular basis of RIT-induced tumor regression may be gained by further studies using different radiation doses. PMID- 7493358 TI - Selection of tumor-specific epitopes on target antigens for radioimmunotherapy of breast cancer. AB - Evidence is presented for two different breast epithelial antigens that some epitopes have greater tumor specificity and are more effective targets for radioimmunotherapy than others. The two antigens, which are major components of the human milk fat globule membrane, are breast mucin and a M(r) 46,000 glycoprotein (BA46). Of five monoclonal antibodies (Mc5, Mc1, BrE-1, BrE-2, and BrE-3) against breast mucin, all recognize overlapping amino acid epitopes on the tandem repeat domain. However, each have unique and different tissue and tumor specificities and unique epitope structures on the fully glycosylated breast mucin. In preclinical studies, radioimmunoconjugates of all five monoclonal antibodies inhibit growth of transplantable breast tumors in immunodeficient mice. In human clinical trials, radioiodinated Mc5 was very poor in localizing breast tumor metastases. On the other hand, 111In-labeled BrE-3 imaged almost 90% of breast tumors and showed promise in radioimmunotherapy when labeled with 90Y. The failure of Mc5 in clinical trials may be partly attributed to the high levels of its epitope on circulating mucin compared to the epitope of BrE-3. The Mc5 binding affinity increased significantly with glycosylation, while the BrE-3 epitope was masked by glycosylation. The BA46 glycoprotein is a breast tumor associated membrane antigen containing an NH2-terminal, epidermal growth factor like domain into which a cell adhesion sequence (RGD) is inserted and a COOH terminal domain with homology to the phospholipid binding C1/C2 domain of coagulation factors V and VIII. It promotes cell attachment in an RGD-dependent manner. Monoclonal antibody Mc8, which binds to the C2-like domain, is only moderately effective in experimental radioimmunotherapy, while Mc3, which binds an epitope in the EGF-like RGD domain, was highly effective in destroying breast tumors in nude mice. With 90Y-labeled Mc3, 6 of 7 mice are cured of the tumors. These results indicate that by selecting appropriate monoclonal antibodies, a normal antigen can be used as a target for radioimmunotherapy. PMID- 7493357 TI - Radioimmunotherapy of breast cancer xenografts with monoclonal antibody ICR12 against c-erbB2 p185: comparison of iodogen and N-succinimidyl 4-methyl-3-(tri-n butylstannyl)benzoate radioiodination methods. AB - C-erbB2 p185 is a proto-oncogene product expressed in 25-30% of human invasive breast cancers that is associated with poor prognosis and resistance to endocrine therapy and chemotherapy. It is minimally expressed in normal adult tissues (M. F. Press et al., Oncogene, 5: 953-962, 1990). For this reason, it is an attractive target for radioimmunotherapy and other antibody-directed therapies. ICR12 is a rat IgG2a monoclonal antibody directed against a protein epitope of the external domain of the c-erbB2 p185. We performed experiments to optimize the direct iodination of ICR12 with 131I using the IodoGen method, and we found impairment of immunoreactive fraction with increasing specific activity. N Succinimidyl 4-methyl-3-(tri-n-butylstannyl)benzoate (MATE) is a tin ester that can be radioiodinated easily and then coupled to the epsilon-amino group of lysine residues. This method has been shown to have improved uptake in tumors compared with antibody labeled by direct iodination (P. K. Garg et al., Nucl. Med. Biol., 20: 379-387, 1993). ICR12 could be labeled up to 16 mCi/mg by this technique without loss of immunoreactive fraction. Whole-body retention of MATE labeled ICR12 was less than IodoGen (P < 0.0001). Radioimmunotherapy experiments in athymic mice bearing established MDA MB 361 human breast cancer xenografts showed growth inhibition for > 24 days at a dose of 600 microCi/mouse (P < 0.0001) when labeled by the IodoGen technique, and 12 days using the MATE method (P < 0.0001). PMID- 7493360 TI - In vitro and in vivo reactivity of an internalizing antibody, RS7, with human breast cancer. AB - RS7, a murine IgG1 antibody raised against human lung carcinoma, possesses pancarcinoma reactivity. The antigen defined by this antibody is present in tumors of the lung, stomach, bladder, breast, ovary, uterus, and prostate. Efficient targeting and therapy by radiolabeled RS7 has been demonstrated previously in animals bearing Calu-3 (an adenocarcinoma of the lung) xenografts. In this study, the efficiency of tumor targeting and the efficacy of therapy of this antibody in nude mice bearing the MDA-MB-468 human breast carcinoma were evaluated. The tumor:nontumor ratios of RS7 were 1.9-2.1 times higher than those for Ag8 (the control antibody) on day 14, except for the heart. These values were similar to that of RS7 in the Calu-3 xenograft model. Radioimmunotherapy using 250 microCi of 131I-labeled RS7 in animals bearing approximately 0.1 cm3 tumors (approximately 10 days old) caused the disappearance of tumors in 6 of 10 animals at 2 weeks postinjection. Tumors eventually disappeared from all animals, and animals remained tumor-free until the termination of the study (11 weeks of duration), except for one animal that developed a transient reappearance of tumor. The tumors in animals that received an equal dose of 131I-labeled Ag8, or unlabeled RS7 or Ag8, either were unchanged or continued to grow. Treatment that used 275 microCi of 131I-labeled RS7 in animals carrying established tumors (1 month old, approximately 0.2-0.3 cm3) showed that this antibody is effective in controlling the growth of this tumor. Tumors in the treatment group began to disappear between the second and third weeks after the injection of the radiolabeled antibody. Seven of 10 animals remained tumor free at 15 weeks after the injection. Tumors in animals that received an equal dose of control antibody persisted but grew at a slower pace compared to the untreated group. No systemic toxicity was observed. PMID- 7493359 TI - Biological activity of two humanized antibodies against two different breast cancer antigens and comparison to their original murine forms. AB - We have humanized two monoclonal antibodies (MoAbs), hu-BrE-3 and hu-Mc3, that are bound to two different antigens of the breast epithelial cell. They bind to the breast epithelial mucin (M(r) 400,000) and the BA46 antigen (M(r) 46,000). They could participate in a joint radioimmunotherapy strategy administering repeated or fractionated dosages, where increased irradiation could be delivered by their simultaneous administration. Both antibodies, hu-BrE-3 and hu-Mc3, had similar reactivity to their antigens and similar binding affinity as those of their original murine forms. However, because humanized MoAbs could have different pharmacokinetic and radioimmunotherapeutic characteristics than their original murine forms, the experimental biodistribution in vivo of both of these two humanized anti-breast tumor MoAbs was compared to their original murine forms. Biodistributions in immunodeficient mice grafted with transplantable human breast tumors, both after radioiodination and 111In labeling via 1-p isothiocyanatobenzyl-methyl-diethylene-triaminepenta-ace tic acid (MXDTPA), demonstrated comparable tumor:normal tissue ratios for the humanized and murine forms. In radioimmunotherapy, the humanized forms for both MoAbs showed also similar tumoricidal activity as that of the original murine MoAbs. These results show that the new humanized forms are amenable to conjugation and radioisotope labeling without loss of biological activity. Furthermore, they demonstrate that these engineered molecules kept intact, both qualitatively and quantitatively, their binding ability, pharmacokinetics, and radioimmunotherapeutic characteristics after the humanization process. PMID- 7493361 TI - Development and characterization of anti-renal cell carcinoma x antichelate bispecific monoclonal antibodies for two-phase targeting of renal cell carcinoma. AB - To test a two-step approach for radioimmunotargeting of renal cell cancer, quadroma cells secreting antichelate x anti-renal cell carcinoma bispecific antibodies were obtained by somatic cell fusion. Five monoclonal antibodies against the chelate 1,4,7-triazaheptane-N,N',N"-pentaacetic acid (DTPA) were produced and characterized. Competitive binding assays indicated that the anti DTPA antibodies reacted with DTPA chelated with indium, yttrium, chromium, iron, or zinc. The affinity constants of the anti-DTPA antibodies for 111In-DTPA ranged from 0.19 to 0.23 nM-1. Using different chelates, a remarkable chelate specificity of the anti-DTPA antibodies was demonstrated. The chelates recognized by the antibodies DTIn1, DTIn2, and DTIn4 share a N(N")-diacetic acid group, whereas the chelates recognized by DTIn3 share a N'-acetic acid group, suggesting the presence of different essential structures within the DTPA molecule that determine the reactivity of the antibodies. Five anti-DTPA antibody-producing hybridomas were used for somatic cell fusion with hybridoma G250 directed against renal cell carcinoma, resulting in three bispecific antibody-producing quadroma cell lines. The bispecific monoclonal antibodies were purified from ascites fluid using protein A affinity chromatography followed by hydroxylapatite chromatography and/or cation exchange chromatography. Of the total IgG amount present in the ascites fluid, 10-15% represented the bispecific antibodies. These bispecific antibodies will allow testing and optimization of a two-step approach for radioimmunotargeting of chelated radionuclides. PMID- 7493362 TI - Use of anticytokeratin monoclonal anti-idiotypic antibodies to improve tumor:nontumor ratio in experimental radioimmunolocalization. AB - A syngeneic, high-affinity, anti-idiotypic monoclonal antibody (MAb; alpha TS1) raised against an anticytokeratin monoclonal antibody (TS1) was evaluated as a second antibody to promote the rapid clearance of radiolabeled TS1 from the blood during experimental radioimmunolocalization. By using a novel biosensor technology (BIAcore), association rate dissociation rate, and affinity constants between the idiotype and the anti-idiotype could be determined. The in vivo results in nude mice carrying HeLa Hep 2 tumors demonstrate the possibility of selectively regulating the amount of the idiotypic 125I-labeled circulating MAb by in vivo injection of this high-affinity, anti-idiotypic antibody. Injection of the anti-idiotype in a molar ratio of 0.75 to the idiotype cleared the blood pool from circulating radiolabeled idiotype within 24 h, with a concomitant rapid excretion of 125I in urine. The total amount of remaining radioactivity in the animals decreased to 15-20% during these 24 h, with the tumors still retaining 60 65% of their initial radioactivity. This approach, using syngeneic primary and secondary MAbs with minimized immunogenicity, significantly improves the tumor:nontumor ratio, thus improving efficiency in experimental radioimmunolocalization and radioimmunotherapy, leaving the endogenous antibody repertoire of the host unaffected. PMID- 7493363 TI - Enhanced radioimmunotargeting of 125I-labeled L6-biotin monoclonal antibody (MAb) by combining preload of cold L6 MAb and subsequent immunoadsorption in rats. AB - The present study investigates whether tumor:normal tissue uptake ratios of radiolabeled monoclonal antibodies can be further improved by a combination of extracorporeal immunoadsorption (ECIA) and preload with unlabeled idiotypic monoclonal antibody. Athymic rats, heterotransplanted with human lung carcinoma under the kidney capsule (SR tumor) and i.m. (IM tumor), were divided into four study groups: controls, ECIA, preload, and combined preload+ECIA. The preload+ECIA procedure reduced the whole-body and plasma activity by 48 and 89%, respectively. After such combined procedure, the uptake of 125I-labeled L6-biotin in SR tumors was unchanged, while the uptake in normal tissues was considerably reduced. Tumor (T):bone marrow ratio was then increased by 17.5 times (after ECIA) and by 4.5 times (24 h after ECIA). Similar enhancements were achieved for T:liver and T:kidney ratios. For the IM tumors, the ratios were not as high as for SR tumors. The effects on T:normal ratios of preload+ECIA in combination were synergistic. The combined procedure resulted both in an increased uptake and prolonged persistence of 125I-labeled L6-biotin in the SR tumors and also in a reduction of corresponding uptake values in organs critical for radiation. PMID- 7493364 TI - Fractionated radioimmunotherapy of human colon carcinoma xenografts with 131I labeled monoclonal antibody CC49. AB - Radiolabeled monoclonal antibodies (MoAbs) have been used for radioimmunotherapy (RIT) of human colon cancer xenografts in an attempt to develop promising clinical approaches to improving therapy success. Several strategies have been investigated to accomplish this task while decreasing toxicity. The CC49 antibody was selected for the present studies because of its relatively high affinity constant (16.2 x 10(9) M-1) for the high molecular weight TAG-72 mucinous antigen secreted by human colon cancer cells. In previous studies, when CC49 was labeled with 131I, it demonstrated a substantial therapeutic advantage over the lower affinity antibody (B72.3) reactive with TAG-72. One of the chief problems in achieving cures with RIT is that hematological toxicity limits the quantity of radionuclide that can be administered. In other studies of dose fractionation using athymic nude mice and 131I-labeled intact MoAbs reactive with human colon cancer xenografts, multiple administrations at approximately 1-week intervals were found to produce more prolonged tumor growth inhibition and less toxicity than single administrations. The purpose of this study was to investigate the therapeutic efficacy and toxicity of 131I-labeled CC49 MoAb administered with short fractionation schedules against human colon cancer xenografts to determine the optimal treatment schedule, with the ultimate aim of applying this approach in clinical trials. The results reported here demonstrate that in an animal colon cancer xenograft model, RIT delivered in a fractionated schedule clearly presents a therapeutic advantage. For example, one administration of 600 microCi 131I labeled CC49 to LS174T tumor-bearing mice was lethal to approximately 25% of mice but produced no tumor regressions. Fractionation of this dose to two administrations of 300 microCi 131I-labeled CC49 at a 3-day interval resulted in tumor regression in approximately 30% of the animals, accompanied by a similar 25% death rate. The administration of 300 microCi 131I-labeled CC49 at a 7-day interval resulted in 15% animal lethality, but no complete tumor regressions. When three administrations of 300 microCi 131I-labeled CC49 were given over a 1 week period on days 0, 3, and 7, tumor regressions occurred in approximately 40% of the animals, accompanied by a 30% death rate. Moreover, three administrations of 300 microCi 131I-labeled CC49 resulted in 20% tumor recurrence, whereas two administrations of 131I-labeled CC49 resulted in 60% tumor recurrence.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 7493365 TI - Review of five consecutive studies of radiolabeled immunoglobulin therapy in Hodgkin's disease. AB - Recurrent Hodgkin's Disease (HD) provides unique opportunities to improve radiolabeled immunoglobulin therapy (RIT). Normal tissue toxicity after RIT is limited to bone marrow damage and is well documented and quantified in HD patients. Anti-antibody formation is rare in patients with HD, allowing for multiple RIT cycles. Overall, 134 patients with recurrent HD were treated on five different studies with i.v. antiferritin, labeled with 131I or with 111In for diagnostic purposes and 90Y for therapeutic purposes. Patients with recurrent, end-stage HD obtain a 60% response rate following 90Y-labeled antiferritin. One half of the therapy responses are complete. Responses are more common in patients with longer disease histories (> 3 years) and smaller tumor volumes (< 30 cm3) and in patients receiving at least 0.4 mCi 90Y-labeled antiferritin/kg body weight. Complete responders survive significantly longer than partial responders (2 years versus 1 year). Partial responders survive longer than patients with progressive disease (1 year versus 4 months). HD in one-third of the patients recurs in new areas. A low protein dose (2-5 mg) and a moderate specific activity (10 mCi/mg) are recommended. Results obtained with 90Y-labeled antiferritin are significantly better than results with 131I-labeled antiferritin. Further translational research in vitro in the radio pharmacy and in vivo with experimental animals is ongoing to improve the therapeutic results of RIT in HD. Obviously, many permutations of RIT cannot be explored in HD patients for ethical, financial, or logistic reasons, and predictive preclinical research is required to achieve further progress. Currently, RIT is a low-toxicity, low-cost outpatient procedure for recurrent HD with a high response rate in a patient population with an unfavorable prognosis. PMID- 7493366 TI - Prognostic factors for radioimmunotherapy in patients with B-lymphocytic malignancies. AB - The Ann Arbor staging classification has proven less useful in nonHodgkin's lymphoma, because this malignancy is inherently a multifocal disorder. Since 1985, 57 adult patients with advanced B-lymphocytic malignancies that progressed despite standard therapy entered into one of three different therapy trials using radiolabeled Lym-1 antibody. Tumor regression in 31 (54%) of these patients fulfilled conventional requirements for an oncological response to the therapy. To define the role of radioimmunotherapy in B-lymphocytic malignancies better and to find opportunities for improving its therapeutic efficacy, the records of these patients were reviewed to assess the significance of various parameters as prognostic indicators. Twenty-one pretherapy characteristics were evaluated, including age at diagnosis, age at study entry, sex, Karnofsky performance status, prior chemotherapy and radiation therapy, interval since diagnosis, histology, constitutional B symptoms, extranodal malignancy (excluding marrow), bone marrow malignancy, tumor bulk, and circulating malignant cells; blood tests included lymphocyte, granulocyte, platelet, hematocrit, serum lactate dehydrogenase (LDH), interleukin 2 receptor, and human antimouse antibody levels. In the multivariate analysis, LDH and Karnofsky performance status were the parameters that best predicted survival, complete and partial remission, and time to progression; interleukin 2 receptor and LDH best predicted complete remission. These prognostic factors for radioimmunotherapy outcome are consistent with the pretherapy characteristics observed to be significant for chemotherapy. PMID- 7493367 TI - Treatment of non-Hodgkin's lymphoma with radiolabeled murine, chimeric, or humanized LL2, an anti-CD22 monoclonal antibody. AB - LL2 is a murine IgG2a anti-CD22 monoclonal antibody found to react with virtually all non-Hodgkin's lymphomas (NHLs). Twenty-one patients with chemotherapy resistant NHL received nonmyeloablative doses of 131I-labeled LL2 IgG and F(ab')2 ranging from 15 to 343 mCi given in cycles of 15-50 mCi, for up to seven treatment cycles. The cumulative protein dose ranged from 1.1 mg IgG to 157 mg F(ab')2. Seventeen patients were assessable for treatment response, and antitumor effects were seen in five (one complete remission, two partial remissions, and two minor or mixed responses). In addition, one complete response was seen in a patient who received only "diagnostic" doses of 131I-LL2 IgG. Thus, a total of six patients had responses according to the defined response criteria. Three additional patients have been treated with potentially myeloablative doses of 131I-LL2 IgG at a starting dose level of 90 mCi/m2 (100 mg). Two patients were evaluable, and both had partial remissions lasting 8 and 3 months, respectively. Chimeric and complementarity-determining region-grafted LL2 have been developed. Initial clinical studies have shown that these agents have targeting properties similar to the murine LL2 and, therefore, may be suitable alternatives to murine LL2 in the treatment of NHL. LL2 is a promising agent for the treatment of lymphoma, particularly when the maximum tolerated dose is given either with or without autologous bone marrow transplantation. PMID- 7493368 TI - Radiolabeled anti-CD33 monoclonal antibody M195 for myeloid leukemias. AB - M195, a mouse monoclonal antibody reactive with the early myeloid antigen CD33, has been shown to target leukemia cells in patients and to reduce large leukemic burdens when labeled with 131I. A complementarity-determining region-grafted, humanized version (HuM195) has demonstrated similar targeting of leukemia cells without immunogenicity. We have studied two applications of therapy with 131I M195. First, to intensify therapy prior to bone marrow transplantation (BMT), we combined 131I-M195 with busulfan and cyclophosphamide. Fifteen patients received first BMT for relapsed or refractory acute myelogenous leukemia or accelerated or blastic chronic myelogenous leukemia; four received second BMT for relapsed chronic or accelerated chronic myelogenous leukemia. Doses of 131I-M195 ranged from 120 to 230 mCi/m2. Few toxicities could be attributed to 131I-M195 therapy, and all patients engrafted. Eighteen patients achieved complete remission. Among those patients receiving first BMT, three have remained in unmaintained remission for 18+ to 29+ months. Six patients relapsed, including one with isolated central nervous system disease 32 months after BMT. Ten patients died in complete remission of transplant-related complications. Second, we studied whether 131I M195 could reduce minimal residual disease and prolong remission and survival durations safely in patients with relapsed acute promyelocytic leukemia after they attained remission with all-trans-retinoic acid. Seven patients were treated with either 50 or 70 mCi/m2 131I-M195. Toxicity was limited to myelosuppression. As a measure of minimal residual disease, we monitored PML/RAR-alpha mRNA by reverse transcription PCR. Six patients had positive reverse transcription PCR assays prior to receiving 131I-M195; two converted transiently to negative. Median disease-free survival and overall survival of the seven patients were 8 (range, 3-14.5) months and 28 (range, 5.5-43+) months, respectively. This regimen compares favorably with others for relapsed acute promyelocytic leukemia. In an effort to avoid nonspecific cytotoxicity associated with 131I in future trials for minimal residual disease, we have conjugated short-range, alpha particle emitting radioisotopes to HuM195 using a bifunctional chelate, 2-(p isothiocyanatobenzyl)-cyclohexyldiethyl-enetriaminep entaacetic acid, with high efficiency and specific activities. 212Bi-HuM195 has demonstrated dose- and specific activity-dependent killing of HL60 cells in vitro. Injection of 213Bi HuM195 into healthy BALB/c mice produced no effects on weight or viability. PMID- 7493370 TI - Radioimmunotherapy for breast cancer using escalating fractionated doses of 131I labeled chimeric L6 antibody with peripheral blood progenitor cell transfusions. AB - Radioimmunotherapy (RAIT) using a humanized murine monoclonal antibody, chimeric L6 (ChL6), has produced objective tumor reduction in 50% of chemotherapy refractory patients with metastatic breast cancer in our prior studies. Because myelosuppression limited dose escalation, we evaluated the ability of granulocyte colony-stimulating factor (G-CSF)-mobilized peripheral blood progenitor cell (PBPC) transfusions to ameliorate this problem. 131I-labeled ChL6 was given at a starting dose of 150 mCi/m2 (2,5 times the maximum tolerated dose without PBPCs) for a planned three treatments. When blood radioactivity declined to less than 1 microCi/ml after treatment, PBPCs were transfused, and G-CSF was administered. Patient 1 had minimal myelosuppression, received two cycles of therapy, and then developed human antimonoclonal antibody (HAMA). Patient 2 had prolonged thrombocytopenia that resolved after additional PBPC transfusion. Progressive disease as well as HAMA prevented further treatment. Patient 3 received all three cycles of 150 mCi/m2 at 8-week intervals. Thrombocytopenia (< 25,000/microliter) occurred but was transient (0-7 days). Because HAMA developed in all prior patients who received G-CSF with ChL6 RAIT, including patients 1 and 2, who received PBPC, patient 3 was given cyclosporin for 14 days. She did not develop HAMA or significant toxicity following 3 cycles of RAIT. Cumulative radiation doses to her lungs and tumor were estimated at 3,100 and 11,200 cGy, respectively. For 9 months, she had a reduction in bone pain, a decline in serum tumor markers, and decreased tumor uptake of F-18-deoxyglucose on a positron emission scan. Her performance status improved, and she had no pulmonary toxicity. We conclude that: (a) PBPC transfusion can modify the myelotoxicity of RAIT and can permit repetitive dosing; (b) cyclosporin is a promising means to abrogate HAMA; and (c) fractionation of intensive-dose RAIT may increase the antitumor effect and reduce normal organ toxicity. PMID- 7493369 TI - Initial tumor targeting, biodistribution, and pharmacokinetic evaluation of the monoclonal antibody PAM4 in patients with pancreatic cancer. AB - This pharmacokinetic study was performed to assess the potential usefulness of the murine monoclonal antibody (MoAb) PAM4-IgG1 as an immunotargeting agent for pancreatic cancer imaging or therapy. This MoAb reacts specifically with mucin purified from human pancreatic cancer. 131I-labeled PAM4-IgG1 was injected i.v. into five patients with suspected pancreatic cancer. Whole-body scans and spot views of the abdominal area were recorded with a computerized gamma camera, and specific regions of interest were drawn over the liver and spleen to define the kinetics of activity in these organs. Blood samples taken from 0.1-144 h after injection served to define the kinetics of plasma distribution and removal of activity from the body. Surgery confirmed pancreatic cancer in four of the five patients, whereas chronic pancreatitis was present in the fifth patient; in all four pancreatic cancer patients, immunostaining with the MoAb PAM4 demonstrated the presence of the specific antigen, with a cytoplasmic and endoluminal/secretory pattern of distribution. Nonspecific radioactivity accumulation in the liver, spleen, and bone marrow was low, linked essentially to the blood pool effect of circulating activity in these organs. The overall quality of scintigraphic maps recorded over the abdomen was quite satisfactory due to the low liver and spleen activity, with good scintigraphic demonstration of the pancreatic cancers (either primary or metastatic); the patient subsequently found to have pancreatitis failed to show PAM4 targeting. Except in one patient with widespread peritoneal metastases (in whom these tumor implants were detected scintigraphically already 24-48 hours after tracer injection), scintigraphic evidence of the tumor lesions was usually late, starting at about 72-96 h after tracer injection. The results obtained in this preliminary study indicate the potential usefulness of MoAb PAM4 for immunoscintigraphy in patients with either primary and/or recurrent pancreatic cancer while also suggesting that the use of the faster-clearing Fab fragments of this MoAb probably would result in improved immunoscintigraphic properties. PMID- 7493371 TI - High-dose 90Y Mx-diethylenetriaminepentaacetic acid (DTPA)-BrE-3 and autologous hematopoietic stem cell support (AHSCS) for the treatment of advanced breast cancer: a phase I trial. AB - This Phase I trial explores the use of high-dose 90Y conjugated to the antibreast cancer monoclonal antibody BrE-3 and autologous hematologic cell support in the treatment of women with stage four breast cancer. Nine women with heavily pretreated disease were enrolled. All of the patients had BrE-3-positive tumors by immunostaining and were treated with increasing doses of 90Y (15 mCi/m2, 3 patients), 20 mCi/M2 (six patients), and a fixed (50 mg) dose of BrE-3. 111In labeled BrE-3 (5 mCi) was given simultaneously for scanning purposes. The only toxicity noted was hematological. Grade 4 platelet toxicity requiring transfusion support occurred in four patients. Grade 4 WBC toxicity was seen in two patients that resolved in 3-9 days. All hematological nadirs occurred approximately 25 days after treatment. Objective partial responses were noted in 4 of 8 (50%) patients with measurable tumors. Dose escalation is ongoing. PMID- 7493372 TI - Enhanced TAG-72 expression and tumor uptake of radiolabeled monoclonal antibody CC49 in metastatic breast cancer patients following alpha-interferon treatment. AB - The IFNs, alpha and gamma, have been shown to enhance the tumor-associated glycoprotein (TAG-72) on adenocarcinoma cells in vitro and in mice with human breast cancer xenografts, resulting in improved targeting of monoclonal antibody CC49. To determine the effect of IFN-alpha on biodistribution and tumor uptake of 131I-labeled CC49, patients with metastatic breast cancer were randomized to either receive or not receive IFN-alpha (3 million units daily for 14 days) by s.c. injection. Three days after beginning IFN-alpha, all patients received 10-20 mCi of 131I-CC49 (specific activity, 16.7 mCi/mg) i.v. Total-body Anger camera scans, along with total-body blood and plasma pharmacokinetics, were performed. Tumor biopsies were taken in all patients before and 48 h after IFN-alpha treatment. There were no significant differences in number of metastases imaged or whole-body, blood and plasma pharmacokinetics between IFN-alpha-treated and untreated patients. Quantitative immunohistochemistry on biopsy specimens from IFN-alpha-treated patients demonstrated a significant increase in mean +/- SEM TAG-72 expression (45.7 +/- 19.4%) compared to patients that were not given IFN alpha (1.3 +/- 0.95%; P < 0.05). Although slight increases in the percent injected dose of 131I-CC49 in tumor occurred after IFN-alpha-treatment, the changes were not significant at the P < 0.05 level. These data suggest that IFN alpha may be useful in enhancing TAG-72 antigen expression in vivo in humans, despite modest improvement in tumor uptake of CC49, possibly because of limited tumor access or other unknown factors. PMID- 7493373 TI - Initial experience evaluating 90yttrium-radiolabeled anti-carcinoembryonic antigen chimeric T84.66 in a phase I radioimmunotherapy trial. AB - Chimeric T84.66 (cT84.66) is a high-affinity (5 x 10(10) M-1) anti-carcino embryonic antigen (CEA) IgG1. In a recently completed pretherapy imaging trial, 111In-labeled cT84.66 demonstrated targeting of CEA-producing metastatic sites and low immunogenicity, with human antichimeric antibody (HACA) response in only 1 of 15 patients after a single administration. The purpose of the present study was to evaluate cT84.66-diethylenetriaminepentaacetic acid labeled with 90Y in a dose-escalation Phase I trial. Patients with metastatic CEA-producing malignancies received imaging doses of 5 mCi 111In-labeled cT84.66 first, followed 1-2 weeks later by 5 mg cT84.66 labeled with the therapeutic dose of 90Y. Immediately following the therapeutic infusion, diethylenetriaminepentaacetic acid was administered by continuous i.v. infusion over 3 days at 250 mg/m2 body surface area/24 h. Biodistribution, tumor targeting, absorbed radiation dose estimates, antibody clearance, and HACA response were evaluated through blood samples, 24-h urine collections, and nuclear images performed at serial time points after infusion. To date, three patients with metastatic colorectal cancer have been evaluated at the first dose level of 5 mCi/m2. No side effects were associated with antibody administration. Localization of the antibody to nonhepatic metastatic sites was observed. Size exclusion high-performance liquid chromatography demonstrated the formation of CEA:antibody complexes in serum in all three patients. A significant variation among patients in the clearance rate of the antibody and complexes from blood to liver was seen, which resulted in a reciprocal relationship between estimated liver dose and red marrow dose. Patients who demonstrated faster clearance to liver demonstrated greater excretion of a low-molecular-weight metabolite through the urine. Two patients developed HACA response, which persisted at 4 months after therapy. At this first dose level, no tumor responses were seen and reversible grade 1 thrombocytopenia was observed in 2 patients. cT84.66 demonstrated effective localization in CEA-producing tumors. Its low immunogenicity after a single administration makes it attractive for further evaluation as a radioimmunotherapeutic agent. However, further evaluation is needed to determine whether its immunogenicity will remain low after multiple administrations. Additionally, in two of the three patients, we identified rapid clearance of the antibody to the liver. This underscores the need to identify, characterize, and understand further those factors that influence the biodistribution and clearance of anti-CEA antibodies to allow for better selection of patients for therapy and rational planning of radioimmunotherapy. PMID- 7493374 TI - Evaluation of a complementarity-determining region-grafted (humanized) anti carcinoembryonic antigen monoclonal antibody in preclinical and clinical studies. AB - A complementarity-determining region-grafted (humanized) version of MN-14 (hMN 14), a high-affinity, anti-carcinoembryonic antigen (CEA) murine monoclonal antibody (mMAb), was selected from several clones that differed slightly in their framework composition. One clone was selected based on its similar binding affinity to CEA as that observed with mMN-14 MAb and its production yields. Targeting studies, using 131I-labeled humanized MN-14 (hMN-14)/125I-labeled mMN 14 IgG in GW-39 tumor-bearing nude mice, showed excellent tumor uptake and tumor: nontumor ratios, similar to the mMN-14. A pilot clinical imaging trial was initiated to determine the targeting, pharmacokinetics, and dosimetry for 131I labeled hMN-14 IgG. Nineteen patients with advanced CEA-producing tumors were given 8 to 30 mCi (0.5 to 20.0 mg). Eleven patients also received 131I-labeled mMN-14 IgG for comparison. The biodistribution, tumor targeting, and pharmacokinetic behavior of the hMN-14 was similar to that seen with the mMN-14. The average time required to clear 50% of the radiolabeled hMN-14 from the blood and total body was 32.9 +/- 25.6 h and 109 +/- 73 h, respectively. Patients with elevated plasma CEA (i.e., > 200 ng/ml) had more than 30% of the labeled antibody complexed within 1 h after injection. In some of these patients, increased complexation resulted in enhanced metabolism of the antibody with more rapid clearance from the blood than that seen in patients with lower plasma CEA. The average radiation absorbed dose measured in 20 tumors (average weight, 204 +/- 205 g) in 14 patients was 7.6 +/- 5.3 cGy/mCi. Tumor: nontumor dose ratios were 2.5 +/- 1.6, 9.5 +/- 5.8, and 2.6 +/- 1.8 for the red marrow, total body, and liver, respectively. One patient, with a highly elevated human anti-mouse antibody response from a prior OncoScint study (murine B72.3 IgG), received 3 injections of the hMN-14 without an adverse experience, and showed no evidence of altered biodistribution characteristic of mMAb-human anti-mouse antibody interactions. An antibody response to hMN-14 (HAhMN14) was not detected in patients who received only the hMN-14 (as many as three injections), but in three patients who received two injections of the mMN-14, a HAhMN14 response was detected. With similar, excellent targeting properties as the mMN-14 and the potential for reduced immunogenicity, hMN-14 is an attractive candidate for further clinical imaging and therapy applications. PMID- 7493375 TI - Targeting and initial radioimmunotherapy of medullary thyroid carcinoma with 131I labeled monoclonal antibodies to carcinoembryonic antigen. AB - The targeting potential of 131I-labeled NP-4 and MN-14 anti-CEA (carcinoembryonic antigen) monoclonal antibodies (MAbs) was assessed in 19 patients with metastatic medullary thyroid cancer (MTC). Seventeen of these patients also entered pilot radioimmunotherapy studies with nonmyeloablative doses of 131I-anti-CEA MAbs. Tumor targeting was possible in all 19 patients, with an overall lesion sensitivity of 91%. Tumor dosimetry with 131I-MN-14 IgG or F(ab)2 was very favorable, with tumor doses of 14.3 +/- 8.3 cGy/mCi and tumor:red marrow dose ratios exceeding 3:1 for most lesions. Limited antitumor effects lasting up to 26+ months, based on physical exam, tumor markers, computed tomography, or a followup MAb scan, were seen in 5 of 11 assessable patients given relatively low doses of 131I-labeled anti-CEA MAbs. We conclude that anti-CEA MAbs are excellent agents for targeting metastatic MTC. The high tumor uptake of the 131I-anti-CEA antibodies and evidence of tumor response in some patients suggest that radioimmunotherapy with radioiodinated anti-CEA MAbs may be an effective treatment for MTC, particularly when the maximum tolerated dose is given alone or in combination with autologous red marrow or peripheral stem cell support. PMID- 7493376 TI - Local treatment of malignant gliomas by direct infusion of specific monoclonal antibodies labeled with 131I: comparison of the results obtained in recurrent and newly diagnosed tumors. AB - Two murine monoclonal antibodies, BC-2 and BC-4, raised against tenascin and labeled with 131I were infused locally in the site of neoplastic disease by means of a removable (16 patients) or indwelling (34 patients) catheter. Fifty patients bearing a malignant glioma were treated. Twenty-six of these were suffering from recurrent disease; their tumors relapsed within 9 months (median) after treatment. The remaining 24 cases had a newly diagnosed tumor, and local radioimmunotherapy (RIT) was given immediately after surgery and radiochemotherapy. All efforts were made to reduce the tumor before the infusion of the radiopharmaceutical. Therefore, 22 cases with relapsing glioma underwent additional debulking surgery, which led to total or subtotal removal of tumor in 9 of the patients. Altogether, 28 patients had intralesional RIT when the disease was minimal or microscopic. Conversely, 22 cases underwent local RIT with a tumor the diameter of which was > 2 cm. In many cases, the infusions were repeated up to six times to achieve complete destruction of the neoplastic tissue. The local treatment did not give rise to systemic or to cerebral adverse effects. The labeled monoclonal antibodies, given directly in the site of the lesion, concentrated in very high amount in the neoplastic tissue and remained fixed in the target for a long period of time. For these reasons, the radiation dose to the tumor was remarkable (on average > 30,000 cGy/cycle) and consequently led to promising results. The median survival was, in total, 20 months (18 in recurrent tumors and 23 in newly diagnosed lesions). Moreover, median survival was 17 months in patients with bulky tumors (both recurrent and newly diagnosed tumors) and 26 months in patients with minimal or microscopic disease. The median time to progression was 3 months in recurrent and 7 months in newly diagnosed gliomas. Finally, RIT produced 3 CRs (all in recurrent tumors), 6 PRs (4 in recurrent and 2 in newly diagnosed), and 11 stabilizations of disease (4 in recurrent and 7 in newly diagnosed). In 19 cases (13 recurrent and 6 newly diagnosed) the progression of tumor was recorded. Eleven patients (2 recurrent and 9 newly diagnosed) who were treated by RIT when their disease was minimal and nondetectable by radiological methods remained disease-free and were classified as NED. The overall response rate (NED plus CR plus PR) was 40% (34.6% recurrent and 45.8% newly diagnosed).(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 7493377 TI - Biological properties of chimeric domain-deleted anticarcinoma immunoglobulins. AB - CC49 is a second-generation monoclonal antibody (MAb) that has high affinity for the tumor-associated pancarcinoma antigen tumor-associated glycoprotein-72. In clinical trials using gamma scanning, radiolabeled CC49 has facilitated the detection of more than 90% of carcinomas. We report here the development of a constant heavy-chain 2 (CH2) domain-deleted chimeric (c) CC49 MAb by transfecting an expression construct consisting of the CC49 murine variable region and a CH2 domain-deleted human IgG1 constant region into cCC49 kappa producing SP2/0 murine myeloma cells. As determined by SDS-PAGE, the intact cCC49 delta CH2 has a molecular weight of 153,000 and, under reducing conditions, molecular weights of 43,000 and 27,000. The plasma clearance and tumor-targeting properties of cCC49 delta CH2 were evaluated and compared with those of mouse/human chimeric forms cCC49 delta CH1 and intact cCC49. Previous studies have shown that the in vitro antigen-binding properties of cCC49 delta CH1 are similar to those of cCC49. Biodistribution studies reported here, using 131I-labeled cCC49 delta CH1 and 125I-labeled cCC49 in athymic mice bearing human colon carcinoma xenografts, demonstrated that both cMAbs localized to the tumor and cleared from the normal tissues similarly. However, in comparison with 125I-labeled cCC49, 131I-labeled cCC49 delta CH2 localized to tumors earlier and had a significantly lower percentage of the injected dose of cMAb/g (%ID/g) in normal tissues than cCC49. Immunoscintigraphy of 131I-labeled cCC49 delta CH2 and 125I-labeled cCC49 in athymic mice bearing human tumor xenografts demonstrated a clear image of the tumor by 24 h after i.v. administration of the delta CH2 cMAb versus the 72 h required for cCC49. Biodistribution studies using 177Lu-conjugated cCC49 delta CH1 and cCC49 showed no significant difference between the radiolocalization indices (% ID/g in tumor divided by % ID/g in normal tissue). 177Lu-conjugated cCC49 delta CH2, however, had lower % ID/g values in tumor xenografts and lower radiolocalization indices than either 177Lu-conjugated cCC49 delta CH1 or 177Lu conjugated cCC49. Pharmacokinetic studies in non-tumor-bearing athymic mice using cCC49 delta CH1 and cCC49 revealed no significant difference between these cMAbs. However, the plasma clearance of cCC49 delta CH2 in non-tumor-bearing mice was significantly faster than that of cCC49. These results were similar when the cMAbs were labeled with either iodine or lutetium. In nonhuman primates, 131I labeled cCC49 delta CH2 cleared significantly faster than 125I-labeled cCC49. The similar plasma clearance and tumor localization of cCC49 and cCC49 delta CH1 suggest that these two cMAbs may be used in similar clinical settings. However, because of the unique pharmacokinetics and tumor targeting of cCC49 delta CH2 versus cCC49 or cCC49 delta CH1, this chimeric immunoglobulin form may be useful in clinical settings that require efficient tumor targeting and rapid serum and whole-body clearance. PMID- 7493378 TI - Bacterial expression of a kemptide fusion protein facilitates 32P labeling of a humanized, anti-carcinoembryonic antigen (hMN-14) antibody fragment. AB - Despite the potential advantages of 32P over other isotopes for radioimmunotherapy, its development as a therapeutic has been hindered by the difficulty of the labeling chemistry. Recently, a heptapeptide [Kemptide (KPT)] has been chemically conjugated to antibodies, and the conjugates have successfully been labeled with 32P enzymatically by using bovine protein kinase. By using genetic engineering, we have produced a chimera (Fab.KPT) consisting of the Fab' moiety of the complementarity-determining region-grafted anti carcinoembryonic antigen-monoclonal antibody, MN14, and a heptapeptide derivative of KPT (Trp-Arg-Arg-Ala-Ser-Leu-Gly). The recombinant protein was expressed in Escherichia coli as a soluble secretory product. The presence of the KPT derivative downstream of the COOH terminus of the hinge region did not impair the binding affinity of the antibody fragment. The Fab.KPT was enzymatically phosphorylated with 32P by bovine protein kinase, without significant effect on the resultant immunoreactivity; 100% of the 32P-labeled Fab.KPT was complexed with liquid carcinoembryonic antigen. The 32P-labeled humanized MN-14 Fab.KPT is expected to have longer blood circulation half-life, allowing for an improved therapeutic efficacy in radioimmunotherapy. PMID- 7493379 TI - Designing human consensus antibodies with minimal positional templates. AB - A humanized antibody retains from the original murine antibody the variable region amino acid residues that are required for antigen binding. These generally include the grafted complementarity determining regions, as well as a few key framework residues. Although the remainder of the framework sequences are imported from a human antibody, they nevertheless differ at a few positions from the human consensus sequences. These atypical residues, which arose by somatic mutation during the affinity maturation of the chosen human antibody, could elicit an immune response in some of the patients receiving the humanized antibody. Thus, ideally one should, instead, choose human consensus frameworks for humanizing murine antibodies. Because there is a different consensus sequence for each of the subclasses of variable light and heavy chains, a method is needed to choose the most appropriate one. We are developing a minimal positional template for such a purpose. A minimal positional template indicates which positions in the variable region frameworks are absolutely required for maintaining the integrity of the binding domains. Therefore, to choose a human framework for humanization, one screens the available human consensus sequences for those that are most similar to the original murine sequence, specifically at the positions indicated by the template. In the subsequent humanization protocol, one then retains all of the murine residues found in the positions indicated in the template while humanizing the residues found at all other positions. A conservative positional template has been applied to the humanizations of the antibreast epithelial mucin antibodies BrE-3 and KC4-G3 without loss of binding affinity. Now we are using progressive cycles of computer modeling and laboratory testing to develop a minimal template. The first of such cycles produced template B, which has been used successfully in the humanization of the antibreast epithelial antigen BA46 antibody Mc3. This prompted us to design template C, which further approaches the desired minimal template. Future constructs will test the validity of this template as well as the validity of this novel humanization approach. PMID- 7493380 TI - Development and evaluation of the specificity of a rat monoclonal anti-idiotype antibody, WN, to an anti-B-cell lymphoma monoclonal antibody, LL2. AB - Anti-idiotype monoclonal antibodies (Mabs) to mLL2, an anti-B-cell lymphoma and CD22-specific murine IgG2a-kappa Mab, were generated by hybridoma technology from splenocytes of Copenhagen rats immunized with mLL2 F(ab')2. Mab WN, an IgG2a kappa, was selected based on its specific binding to mLL2 and not other IgG isotypes or anti-B-cell Mabs. In a radioimmunoassay, WN was found to inhibit the binding of 125I-labeled mLL2 to Raji cells and to have no effect on the binding of other B-cell-reactive antibodies. Using high performance liquid chromatography analysis, WN was shown to complex specifically with both mLL2 and mLL2 Fab'. Meanwhile, we have constructed chimeric (cLL2) and humanized (hLL2) versions of LL2. Both cLL2 and hLL2 were demonstrated to retain the original antigen specificity and affinity of mLL2 [S.O. Leung et al., Proc. Am. Assoc. Cancer Res., 2872 (abstract), 34: 481, 1993]. The specific binding of WN to either radioiodinated or peroxidase-conjugated mLL2 was inhibited in a dose-response manner, and to a similar extent by mLL2, cLL2, and hLL2. Since the mLL2 complementarity-determining regions are the only sequences common to mLL2, cLL2, and hLL2, the result confirms that WN is specific to the antigen-binding complementarity-determining regions. A WN binding assay is currently being evaluated as a substitute for the tedious, and sometimes inconsistent, Raji cell binding assay for the determination of LL2 immunoreactivity. In conclusion, we have developed an anti-idiotype Mab, WN, to mLL2. Its potential use as a surrogate antigen for B-cell lymphoma is under investigation. PMID- 7493381 TI - Targeting c-erbB-2 expressing tumors using single-chain Fv monomers and dimers. AB - Single-chain Fv proteins containing a COOH-terminal cysteine (sFv') were constructed by using an antidigoxin 26.10 sFv and an anti-c-erbB-2 741F8 sFv. The fully active sFv' proteins were prepared by expression in Escherichia coli as insoluble inclusion bodies, followed by in vitro refolding using glutathione redox buffers and purification. The COOH-terminal cysteines of the refolded sFv' proteins were protected by a blocking group presumed to be the glutathionyl peptide, which was easily and selectively removed by gentle reduction. Air oxidation of the reduced sFv' monomers resulted in the efficient formation of disulfide-linked sFv' homodimers, designated (sFv')2, which were stable under oxidizing conditions and relatively slow to be disrupted under reducing conditions. The (26-10-1 sFv')-(741F8-1 sFv') heterodimer was prepared and possessed dual-antigen specificity; the active bispecific (sFv')2 dimerized under native conditions, apparently as a manifestation of self-association by the 741F8 sFv' subunit. Biodistribution and imaging studies that were performed on mice bearing human SK-OV-3 tumor xenografts that express the c-erbB-2 as a cell surface antigen were reviewed. Radioiodinated 741F8-2 (sFv')2 homodimer localized to the tumors with high specificity, as evidenced by excellent tumor:normal tissue ratios. Sagittal section autoradiography of whole animals 24 h after administration of antibody species revealed that 741F8 (sFv')2 produced a stronger tumor image than comparable doses of the 741F8 Fab, monomeric sFv', and the 26-10 (sFv')2 control without the high nonspecific background distribution of the 741F8 IgG. PMID- 7493382 TI - To delegate or not to delegate--is that the question? PMID- 7493383 TI - PCS nurse associates--created to meet need. PMID- 7493384 TI - Case management and hospice nursing--a good fit. PMID- 7493385 TI - Nurses campaign for adolescent health and prevention of HIV infections. PMID- 7493386 TI - Alternative dispute resolution--issues in health care. PMID- 7493387 TI - Leadership through shared governance. PMID- 7493388 TI - Campaign for employee rights. PMID- 7493389 TI - Beginning a job search? Take six steps to ensure success. PMID- 7493390 TI - Who, what, where news on ethics committees. PMID- 7493391 TI - Retirement planning: you can't afford to neglect it. PMID- 7493392 TI - Techniques for successful grant-writing: Colorado State Veterans Nursing Home. PMID- 7493394 TI - Power and professionalism. PMID- 7493395 TI - Membrane-bound/soluble IL-2 receptor (IL-2R) and levels of IL-1 alpha, IL-2, and IL-6 in the serum and in the PBMC culture supernatants from 17 patients with hematological malignancies. AB - The present study investigated the peripheral blood mononuclear cells (PBMC) blastic responses to PHA, PHA plus recombinant IL-2 (rIL-2) and rIL-2 alone; the expression of membrane-bound IL-2R on PHA-stimulated PBMC; and the levels of IL-1 alpha, IL-2, IL-6, and sIL-2R in serum and in culture supernatants from PHA stimulated PBMC in 17 patients with with non-Hodgkin's lymphoma (NHL), 4 with Hodgkin's lymphoma (HL), 5 with Hairy cell leukemia, 1 with chronic myelogenous leukemia, and 1 with chronic lymphocytic leukemia. The patients with HL and NHL with active disease (AD) were separated from those in clinical remission. The patients with AD were studied at diagnosis (obviously before therapy) and the patients in clinical remission were out of therapy since at least 6 mo. The lymphocyte blastogenic response to PHA was significantly lower in patients with HL and NHL with AD than in the control group. The response to rIL-2 alone was in the same range in the control group and in HL and NHL AD patients. By adding rIL 2 to PHA there was an increase of the blastogenic response of the same patients. The percentage of CD25 expressed on PHA-stimulated lymphocytes from patients with HL and NHL AD and from normal subjects is in the same range. Serum levels of IL 2, IL-6, and sIL-2R were significantly higher in HL and NHL AD patients than in controls as well as in all other hematological malignancies. Supernatants derived from PHA-stimulated PBMC were assessed for the presence of cytokines and sIL-2R by ELISA. The levels of IL-2, IL-6, and sIL-2R were significantly lower in HL and NHL AD patients than in controls as well as in all other hematological malignancies. PMID- 7493397 TI - Syngeneic anti-idiotypic antibodies eliminate excess radiolabeled idiotypes at experimental radioimmunolocalization. AB - Significant improvements in tumor/nontumor ratio can be achieved by injections of nonlabeled anti-idiotypic monoclonal antibodies (MAbs) during radioimmunolocalization and radioimmunotherapy using MAbs to target experimental tumors. The in vivo effects of an anti-idiotypic MAb (alpha H7) against a radioiodinated, high affinity, low dissociation rate, monoclonal antiplacental alkaline phosphatase antibody (H7) was investigated. Following in vivo injection of the anti-idiotypic MAb, the radioactivity in experimental tumors was found to decrease only 25% while the reduction of corresponding radioactivity in nontumor tissues amounted to 65-85%, compared to the group receiving no anti-idiotypic MAbs. These results indicate that it is possible to partially clear the circulation and nontumor tissues from excess of radiolabeled idiotypic antibody, without significant decrease in specific tumor localization, increasing the tumor/nontumor ratio three- to fourfold. Circulating nontumor targeting radiolabeled antibodies is one of the major limiting factors in radioimmunotherapy today. Injection of anti-idiotypic MAbs could selectively significantly reduce the radiation dose to radio-sensitive tissues, i.e., bone marrow and intestine, thus improving efficiency in radioimmunoscintigraphy and radioimmunotherapy. PMID- 7493396 TI - Animal models of human-derived cancer vaccines. AB - Preclinical cancer vaccine studies must address vaccine safety, immunogenicity, and efficacy, as well as mechanism of vaccine action. Animal models of vaccines employing human tumor-associated antigen or epitopes (TAA, TAE) differ fundamentally from those employing tumor-specific antigens or epitopes (TSA, TSE). TSA and TSE vaccines will most likely demonstrate similar toxicity, immunogenicity, and efficacy in both tumor-bearing animals and patients. In contrast, TAA/TAE immunizations may have to overcome a host's immunological tolerance to TAA/TAE expressed not only on tumor, but also on normal tissues; immunity to TAA/TAE will potentially target normal tissues and thus may induce autoimmunity. Various experimental models for human-derived TAA/TAE vaccines have been developed. These models include transgenic mice, mice with severe combined immunodeficiency (SCID), and non-human primates. Recently, unique animal models of TAA/TAE cancer vaccines have been developed, taking advantage of the discovery of animal tissue antigens with significant sequence homologies to human TAA/TAE. These models mimic perhaps most closely the situation in cancer patients. PMID- 7493398 TI - Engineering recombinant antibodies for immunotherapy. AB - Recombinant antibody fragments binding with high affinity to their target can be obtained either from hybridomas or directly from antibody libraries on filamentous phage. These fragments are devoid of any activity other than antigen binding, and have to be processed and functionalized in order to be suitable for clinical applications. This article presents the authors' view on the procedures and the features that are important for effective transformation of recombinant antibodies into useful immunotherapeutic agents. The topics presented include phage display methodologies, engineering of high-affinity binding, purification, and functionalization strategies of recombinant antibodies. PMID- 7493393 TI - Registering as an advanced practice nurse. PMID- 7493399 TI - Cerebellar granule cells in vitro recapitulate the in vivo pattern of GABAA receptor subunit expression. AB - GABAA-receptor heterogeneity is based on the combinatorial assembly of a family of 15 subunits (alpha 1-6, beta 1-3, gamma 1-3, delta, rho 1-2) into multiple receptor subtypes. The factors regulating the differential expression of GABAA receptor subtypes in the CNS are largely unknown. In the present study, we have used primary cultures of rat cerebellar granule cells as model system to analyze to which extent the expression, subunit composition, and subcellular localization of GABAA-receptors depend on signals available in the cerebellum in vivo, in particular GABAergic input which is lost in vitro. GABAA-receptor subunits were visualized immunohistochemically with subunit-specific antibodies and their subcellular localization was investigated by confocal laser microscopy with selective markers for dendritic proteins (microtubule-associated protein 2, non phosphorylated neurofilaments) and synaptic vesicles (synaptophysin). Triple immunofluorescence staining experiments revealed that dissociated granule cells taken from 7-day-old rats and maintained in culture for 9 days co-expressed the subunits alpha 1 alpha 6 beta 2,3 gamma 2 delta, a subunit repertoire which is identical to that seen in vivo in the adult cerebellum. GABAA-receptor subunits were localized selectively in granule cell somata and in proximal neurites immunopositive for MAP-2. No staining was detected in distal neurites immunopositive for neurofilaments. GABAA-receptor subunits frequently were aggregated in clusters closely apposed to synaptophysin-immunoreactive varicosities, suggesting a post-synaptic localization. Thus, major functional determinants of GABAA-receptors in granule cells (subunit repertoire, subcellular segregation and clustering in post-synaptic sites) develop in vitro, indicating that they are regulated to a large extent by intrinsic factors, independently of GABAergic input. PMID- 7493400 TI - Insulin-like growth factor I increases myelination and inhibits demyelination in cultured organotypic nerve tissue. AB - The implication of insulin-like growth factor I (IGF-I) in the myelination and the repair of myelin that occur after a demyelinating process was evaluated in organotypic cultures of embryonic nerve tissue. The amount of myelin of mouse spinal cord explants exposed to media supplemented with IGF-I beginning on the first day of explantation was recorded by light-microscopic examination and quantitation of the 2',3'-cyclic nucleotide 3'-phosphohydrolase (CNPase) activity. After 9 days in vitro (DIV), the cultures treated with medium supplemented with 0.1-1 microgram/ml IGF-I showed a greater amount of myelin and an increase over the controls in CNPase activity between 50 and 80% at 16 DIV and 100% at 21 DIV. Total demyelination with a concomitant reduction of about 80% in the CNPase activity resulted when anti-white matter antiserum and complement were added to the nutrient medium of fully myelinated cultures. This effect was partially reverted when IGF-I was included in the demyelinating medium. The higher inhibition, about 50%, was obtained with concentrations of IGF-I between 0.1 and 0.5 micrograms/ml. To study the effect of IGF-I on remyelination, well myelinated cultures were completely demyelinated, maintained in that state for 2 or 15 DIV and after that allowed to remyelinate for 14 days. Cultures exposed to medium supplemented with 0.01-0.1 microgram/ml IGF-I showed a degree of remyelination similar to that of the normal nutrient medium-fed cultures.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7493401 TI - Somatostatin enhances neurofilament expression and neurite outgrowth in cultured rat cerebellar granule cells. AB - Somatostatin and its receptors are transiently expressed at a high level in the cerebellum around birth, before declining to adult levels by 2-3 weeks postnatally. We therefore investigated the neurotrophic effects of somatostatin (SS) on rat cerebellar granule cells in culture by measuring the percentage of cells with processes, the content of mRNA and protein for neurofilament (NF) and mRNA for glutaminase, and the number of viable cells (MTS assay). SS increased the percentage of cells with processes at 8 h after plating. After 1 day in vitro (DIV), SS caused a 2-fold increase in NF mRNA, and a 23% increase in NF protein. The mRNA increase was maximal at DIV1 whereas by DIV7 the NF protein content of control cells reached that of SS-treated cells. SS had no effect on glutaminase mRNA or on the number of viable neurons from either postnatal day 5 or 8 animals. These results demonstrated that SS has a neurotrophic effect on neurite production, including initiation of neurite outgrowth, but no effect on neuronal survival, cell proliferation, or phenotype differentiation (glutaminase expression), and support the possibility that SS plays a role in the differentiation of immature cerebellar granule cells during central nervous system development. PMID- 7493403 TI - Production of hyaluronan-dependent pericellular matrix by embryonic rat glial cells. AB - The extracellular matrix of brain is largely composed of aggregates formed by assembly of many proteoglycan and link protein molecules along a hyaluronan polymer backbone. Some cell types construct large, highly hydrated, pericellular matrices or 'coats' from these hyaluronan-mediated aggregates. We show here that embryonic glial cells produce such hyaluronan-dependent pericellular matrices in response to addition of serum or basic fibroblast growth factor plus transforming growth factor-beta. It is proposed that such a matrix is a significant component of the extracellular milieu of the brain, especially during morphogenesis within the developing brain, and that basic fibroblast growth factor and transforming growth factor-beta regulate its production. PMID- 7493402 TI - The mitochondrial inhibitor malonate enhances NMDA toxicity in the neonatal rat striatum. AB - Intra-striatal injections of the mitochondrial inhibitor malonate elicit age dependent neuronal damage in rat brain; injury is more extensive in older animals than in young adults. We investigated the neurotoxic potential of malonate in the immature rat brain. We found that 7-day-old (P7) rats were highly resistant to malonate neurotoxicity. Yet, although intra-striatal injections of 1 mumol malonate did not elicit overt tissue injury in P7 rats, co-administration of this dose of malonate with a dose of NMDA close to its toxicity threshold (2.5 nmol) doubled the severity of resulting excitotoxic injury. PMID- 7493404 TI - Neural input and the development of adrenergic intracellular signaling: neonatal denervation evokes neither receptor upregulation nor persistent supersensitivity of adenylate cyclase. AB - In the adult, denervation of adrenergic target tissues leads to compensatory upregulation of receptor sites and to supersensitive responses. When 6 hydroxydopamine (6-OHDA) was given to neonatal rats, cardiac beta-receptors failed to show significant upregulation throughout the first five postnatal weeks and alpha 1-receptors were unchanged except at 35 days of age, despite 70-95% depletion of norepinephrine. The failure to upregulate could not be attributed to the high background level of receptor expression commensurate with ontogenetic increases in receptor numbers, since the same deficiency was seen in the liver, a tissue in which beta-receptors decline with development; liver alpha 1-receptors also failed to upregulate after neonatal denervation. Examination of the linkage of beta-receptors to adenylate cyclase indicated major differences from mature regulatory mechanisms, as denervation supersensitivity was completely absent (liver) or emerged only transiently several weeks after 6-OHDA treatment (heart). In the heart, there was evidence for a defect in the G-protein-dependent component of the receptor/cyclase linkage that could contribute to the delayed appearance of supersensitivity. Because the fundamental patterns of receptor ontogeny and of adenylate cyclase responsiveness are still present after neonatal denervation, it is unlikely that neural input provides the major impetus for basal development. However, adult-type regulation of receptors and responses did not emerge even after a prolonged period; thus, neural input during a critical developmental stage may be required for the cell to learn how to adjust receptor expression and the receptor/cyclase link in response to stimulation. PMID- 7493405 TI - Developmental expression of G alpha o and G alpha s isoforms in PC12 cells: relationship to neurite outgrowth. AB - Recent evidence suggests neuronal differentiation is associated with developmentally regulated changes in the expression of certain G-protein subtypes but the relationship of such changes to neuronal differentiation and/or neurite outgrowth is poorly understood. In this study, the appearance of the G-protein alpha o and alpha s subunit isoforms was characterized in NGF-induced differentiating pheochromocytoma (PC)12 cells and correlated with neurite outgrowth, which was assessed with direct morphometric measures at 24-h intervals for a 5-day period and at 24, 48 and 72 h following NGF removal. Significant increases were observed in alpha o1 immunoreactive levels and the ratio of alpha o1/alpha o2 immunoreactivities during NGF exposure but not in the levels of either the long or short isoform of alpha s. The former changes also correlated significantly with neurite length during NGF treatment. Furthermore, no significant changes were observed in the levels of alpha o or alpha s isoforms following NGF removal. These findings suggest that alpha o isoforms are involved in neuritic extension in PC12 cells. PMID- 7493406 TI - MMP-9 (gelatinase B) mRNA is expressed during mouse neurogenesis and may be associated with vascularization. AB - Expression of MMP-9 mRNA, a type IV collagenase gene product, was followed during embryonic development of the mouse brain using in situ hybridization. Murine embryos from 7.5 to 15 days after fertilization were sectioned and evaluated for MMP-9 expression. During early development, from day 7.5 to day 9, no signal was detected in the cells of the neuroepithelium or in cells of the cephalic mesenchyme of the neural tube. At day 11, gene expression was localized to the Rathke's pouch and the germinal zone of the primitive ventricular system. At day 13, but most notably at day 15, high levels of MMP-9 were expressed by progenitor cells in close association with the development of structures, such as the hypophysis, the choroid plexus, the ganglion cell layer of the retina and the uveal tract. High MMP-9 mRNA levels were also associated with dense cellular aggregates destined to form the highly vascular grey matter of the brain. The presence of MMP-9 mRNA was confirmed using a ribonuclease protection assay. A 105 kDa gelatinase, consistent with the expected molecular mass for the murine MMP-9, was detected in embryonic brain extracts by substrate gel electrophoresis. To our knowledge, this is the first report on the localization of MMP-9 in developing neural tissues. Our results suggest that MMP-9 expression may have a previously unsuspected role in neural development. PMID- 7493409 TI - Distribution of 5'-nucleotidase in the developing mouse retina. AB - The distribution of 5'-nucleotidase was investigated in the developing mouse retina and also in the retina of the adult mouse and rat using a monospecific antibody directed against 5'-nucleotidase isolated from bovine brain. Already at the stage of the formation of the eye cup strong immunofluorescence was found at the ventricular surface. Throughout development the enzyme remained associated with the outer surface of the retina that corresponds to the former inner surface of the eye vesicle. During embryonic development immunoreactivity was also associated with the proliferating cellular elements which at that stage cannot yet be attributed to a defined retinal cell type. In the adult stage the surface located retinal immunoreactivity is assigned to Muller cells with strongest fluorescence at the apical and microvilli containing cell compartment. Also the cellular processes in the outer nuclear layer, the outer plexiform layer and to a minor extent the inner nuclear layer revealed immunoreactivity. 5'-Nucleotidase immunoreactivity was found at the ventricular surface also of the adult brain. There it was associated with the apical surface of ependymal cells. Our results suggest that 5'-nucleotidase is of general functional importance for the metabolism of nucleotides at the ventricular surface of the retina as well as the ventricles of the brain, a feature that is maintained throughout development. Muller cells thus share not only functional characteristics of astrocytes and oligodendrocytes, as previously revealed, but also of ependymal cells to which they are closely related. PMID- 7493410 TI - Expression of the amyloid protein precursor of Alzheimer's disease in the developing rat olfactory system. AB - The expression of the amyloid protein precursor (APP) of Alzheimer's disease (AD) was examined in the olfactory system of the developing rat. Two monoclonal antibodies were used to detect APP: Alz-90, which specifically recognizes APP, and 22C11 which recognizes both APP and the structurally related protein APLP-2. Very similar patterns of immunoreactivity were observed with both antibodies. APP immunoreactivity was first detected in a subpopulation of olfactory epithelial cells at embryonic day 16 (E16), at a time when primary sensory olfactory axons are first beginning to pierce the glia limitans of the olfactory bulb. At E16, there were more olfactory receptor neurons which expressed APP than the olfactory marker protein (OMP), indicating that some APP-containing neurons were not fully mature. Between E16 and postnatal day 8 (P8), there was a marked increase in the number of primary sensory olfactory neurons expressing APP. In the olfactory bulb, APP was first detected in the mitral cell layer at E18, at a time when synapses are first beginning to form between the dendrites of these cells and primary sensory axons. The level of APP detected within mitral cell perikarya decreased after birth and could no longer be detected between P3 and P8. This indicated that once synaptic connections had been initiated within olfactory glomeruli, the expression of APP within the mitral cells was down-regulated. High levels of APP were, however, detected within the olfactory nerve fiber layer and glomeruli between P3 and P8. The results demonstrate that APP expression in the olfactory system is coordinately regulated with the major periods of synaptogenesis.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7493408 TI - Control of myelination, axonal growth, and synapse formation in spinal cord explants by ion channels and electrical activity. AB - The involvement of axonal electrical activity and ion channels as mediators of neuron-glial communication during myelin formation has been tested in explant culture. Transverse slices of embryonic mouse spinal cord were maintained under conditions normally leading to extensive myelination. Axonal conduction was measured optically through the use of a voltage-sensitive dye. Glial development was at a very early stage at the time of plating, and oligodendrocyte precursor cells had not yet appeared. Spontaneous electrical activity was blocked either by tetrodotoxin or by elevation of external K+ concentrations. Myelin development was unaffected by tetrodotoxin and was also present, though quantitatively reduced, in elevated K+. Tetraethylammonium ion (TEA+), a blocker of many K+ channels, almost entirely eliminated myelination at a concentration of 1 mM, but axonal growth and conduction were unaffected. Synapse formation was followed both morphologically and functionally, and was altered neither by conduction block nor by 1 mM TEA+. It is concluded that in the spinal cord oligodendrocyte development and myelination can proceed in the absence of axonal action potentials, but ion channels, possibly in glial membranes, play an important role in these events. PMID- 7493407 TI - Differential distributions of HNK-1 and tenascin immunoreactivity during innervation of myotomal muscle in Xenopus. AB - The HNK-1 antibody labels developing neurons and their processes in Xenopus [40,43]. HNK-1 recognizes a carbohydrate bound to subsets of a number of cell and extracellular matrix (ECM) adhesion molecules, including those of the tenascin/cytotactin family. Both HNK-1 and Tenascin (TN) antigens have been implicated in neural development. Here we describe and compare expression patterns of the two antigens during development of the myotomal neuromuscular system. HNK-1 marks peripheral motor axon processes, including their fine branches at neuromuscular junctions (NMJs), in a pattern that persists throughout development. NMJs of myotomal muscle are located on either side of the intermyotomal spaces (IMS). From the beginning, TN-immunoreactivity (TN-IR) appears predominantly at the IMS, but not on peripheral nerves. Expression of both antigens is graded along the anteroposterior axis, with tenascin appearing first. At the ultrastructural level, HNK1-immunoreactivity appears on motor axons and at the neuromuscular junction on pre- and postsynaptic membranes. In contrast, TN-IR appears ubiquitously in the ECM of the intermyotomal junction, but not in association with axons or synaptic components. The respective distributions of the two antigens during myotomal innervation indicate that tenascin does not bear the HNK-1 antigen in Xenopus embryos and larvae, but that both antigens occupy positions consistent with roles in the establishment of NMJs at the myotomes. PMID- 7493412 TI - Faculty certification? Yes! PMID- 7493411 TI - Developmental expression of endothelin receptors in cerebellar neurons differentiating in culture. AB - We describe the identification and expression of endothelin (ET) receptor subtypes in differentiating cultured cerebellar neurons. Using [125I]ET-1 and the subtype-selective ligands BQ-123 and sarafotoxin 6c as selective ligands for the ETA and ETB receptors, respectively, we found that cerebellum from 8-day-old rats displayed only the ETB receptor subtype. We next cultivated cerebellar granule cell neurons to study ET receptor differentiation between 2 and 22 days in vitro. Using the above reagents, we found that while unlabeled ET-1 displayed monophasic competition curves, BQ-123 and sarafotoxin 6c displayed partial displacement curves, indicating the presence of both ETA and ETB receptors on these neurons. The proportion of ETB receptors gradually decreased from day 2 onwards the proportion of ETA receptors gradually increased. On days 2, 3, 4, and 5 of culture, the ETB:ETA receptor ratios were 90:10, 70:30, 60:40, and 40:60, respectively. There was no further change in receptor subtype ratio beyond day 5 and up to day 22. Northern blot analysis showed that ETB receptor message expression was 6.9-fold higher than that of ETA receptor expression on day 2, but steadily decreased with time, whereas ETA receptor message expression was minimal on day 2 and maximal by day 3 and 4. By day 7, receptor message was of equal abundance, which was in good agreement with the binding studies. This novel, developmentally regulated process predicts the existence of endogenous mediators of neuronal ET receptor expression. PMID- 7493413 TI - Fibrin D-dimer: a useful clinical marker of thrombogenesis? PMID- 7493414 TI - t(8;21) breakpoints are clustered between alternatively spliced exons of MTG8. AB - 1. The (8;21) translocation, which is consistently associated with a subgroup of acute myeloid leukaemia, involves two loci: runt on chromosome 21 and MTG8 on chromosome 8. 2. Breakpoints in runt fall within a single intron that is located immediately downstream of a phylogenetically conserved DNA-binding domain (the 'runt box'). 3. We now show that most breakpoints on chromosome 8 fall within a region between two alternative 5' MTG8 exons. Thus, we predict that chimaeric genes on both the derivative(8) and the derivative(21) chromosomes have the potential to be transcriptionally active. PMID- 7493415 TI - Role of alpha-adrenoceptors in the maintenance of core temperature in humans. AB - 1. Although alpha-adrenoceptor antagonists have been shown to induce core hypothermia in animals, it is unclear whether the primary mechanism is increased heat loss or decreased heat production. Furthermore, studies have not been performed in humans to determine the role of alpha-adrenoceptors in the maintenance of core temperature. 2. alpha-Adrenoceptor blockade was achieved with three doses of phentolamine given by random assignment on three different study days in five male and five female healthy subjects. Core temperature, mean skin surface temperature, fingertip capillary blood flow and metabolic heat production were measured. Dose-response curves were plotted for all measured variables, and males and females were compared to identify potential gender differences. 3. Core temperature decreased with all doses of phentolamine. At the completion of the phentolamine infusion, the decrease in core temperature was more significant with high-dose (0.3 +/- 0.1 degrees C, P = 0.03) and with medium-dose (0.2 +/- 0.0 degrees C, P = 0.05) phentolamine than with low-dose phentolamine (0.1 +/- 0.0 degrees C). The maximum core temperature decrease during the study was more significant with high dose (0.6 +/- 1 degrees C) than with medium (0.3 +/- 1 degrees C, P = 0.04) or low (0.3 +/- 1 degrees C, P = 0.005) doses. Mean skin surface temperature was increased with all doses. Fingertip blood flow was increased (approximately 60% above baseline) with the medium and high doses, but was unchanged with the low dose. Total body oxygen consumption was unchanged regardless of dose.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7493416 TI - Vasodilatory responses of isolated arteries of cirrhotic rats. AB - 1. There is currently considerable interest in the role of locally produced vasodilators such as nitric oxide and adenosine in the pathogenesis of the peripheral vasodilatation of cirrhosis. However, the signal transduction pathways involving guanylate cyclase and adenylate cyclase have not been clearly delineated in the isolated blood vessel. 2. We therefore aimed to examine the in vitro vasorelaxant effects of the endothelium-dependent dilator bethanechol, the endothelium-independent dilator sodium nitroprusside and adenosine, as drugs that work via activation of guanylate and adenylate cyclases, in isolated aortic and superior mesenteric arterial rings from cirrhotic and control rats. 3. Cirrhosis was induced by chronic bile duct ligation and section of 24-28 days' duration, while controls underwent sham operation. The vessels were precontracted with the alpha 1-adrenoceptor agonist phenylephrine, then relaxed by incremental doses of the three drugs. 4. Marked attenuation of vasoconstriction induced by phenylephrine in isolated aortic and mesenteric arterial rings from cirrhotic rats compared with the control vessels was observed. 5. There were no significant differences in relaxation between the cirrhotic and control vessels to the three drugs. We conclude that in vitro vasodilatory responses mediated through signal transduction pathways involving guanylate cyclase and adenylate cyclase remain unchanged in a rat model of biliary cirrhosis. PMID- 7493417 TI - Cutaneous vasoconstrictor response induced by inspiratory gasp in relation to sex and age. AB - 1. Vasoconstrictor responses to inspiratory gasp were determined in fingertip skin by laser Doppler flowmetry in relation to age and gender of healthy adults. Variations of the responses were examined together with variations in body mass index and vital capacity. 2. In the absence of any significant difference in the baseline blood flow, the index of vasoconstrictor response (per cent change) induced by inspiratory gasp was higher in young males than in young females (71.4% versus 59.4%; P < 0.05), and in the same direction as the change in vital capacity determined in the same laboratory setting. Similar parallel differences in the indexes of vasoconstrictor response and vital capacity existed between young and middle-aged males, suggesting that the differences observed were not due to gender or age per se. 3. Strong positive correlations between the index of the vasoconstrictor response and vital capacity in the same subject were found when young males and females were pooled together and when young and middle-aged males were pooled together. Furthermore, after graded inspiratory gasp, graded vasoconstrictor response was observed in the same subject, indicating a close relationship between the depth of inspiration (magnitude of stimulus) and the index of the vasoconstrictor response (reactivity). 4. We concluded that the index of the vasoconstrictor response induced by inspiratory gasp depends significantly on the magnitude of the vital capacity. These results provide a new approach for quantifying dynamic changes in skin blood flow and raise caveats about comparing differences in neurovascular functions using single measurements of vasoconstrictor response induced by inspiratory gasp as an index. PMID- 7493418 TI - Contribution of age and intimal lesion morphology to coronary artery wall mechanics in coronary artery disease. AB - 1. Atherosclerotic changes of the coronary vascular wall include localized plaque formation and diffuse hardening, both of which influence tensile strength of the inner vascular wall. Insight into the relation between age, the atherosclerotic process and vessel wall elasticity may contribute to the prevention of secondary vascular wall damage. 2. To evaluate the determinants of coronary artery wall elasticity, we studied a group of 34 patients, aged 54.6 +/- 10.1 years, who underwent balloon angioplasty for coronary artery disease. With the use of 30 MHz intravascular ultrasound, a total of 127 cross-sections of coronary artery segments outside the balloon area were studied. The echographic appearance of the intima was classified as normal intimal thickening, fibrous lesions and calcified lesions. Fibrofatty lesions, which are believed to be rupture prone, were analysed separately. From the lumen area change, measured by semiautomated planimetry from digitized ultrasound images, and simultaneously measured aortic pressure change during the diastolic phase of the cardiac cycle, two elastic parameters were derived, pressure-strain modulus Ep and beta-stiffness. 3. In univariate analysis Ep and beta-stiffness were determined by age, vessel size, intimal composition, severity of lumen obstruction and gender, but not by eccentricity of the lesion or a specific coronary artery. In multivariate linear analysis, increased vessel wall stiffness was independently determined by advanced age, larger vessel size, intimal composition, and to a lesser extent by female gender. Elastic parameters of fibrofatty lesions were not different from other fibrous lesions. 4. Coronary artery wall elasticity is independently determined by age, vessel size and intimal composition. PMID- 7493419 TI - Aortic distensibility in patients with cerebrovascular disease. AB - 1. Non-invasive aortic compliance measurements have been used previously to assess the distensibility of the aorta in several pathological conditions associated with increased cardiovascular risk. We set out to establish whether aortic compliance is abnormal in patients with stroke. 2. Pulse wave velocity measurements of thoraco-abdominal aortic compliance were made in 20 stroke patients and 25 age- and sex-matched hospitalized, non-stroke control subjects putatively free of cardiovascular disease. Since compliance varies with non chronic changes in blood pressure, a blood pressure corrected index of aortic distensibility, Cp, was calculated. 3. Aortic compliance was significantly reduced in patients with stroke compared with non-stroke control subjects (0.46 +/- 0.27 versus 0.86 +/- 0.34%/10 mmHg, P < 0.0002), corresponding with higher values for pulse wave velocity. Stroke patients also had significantly higher systolic and diastolic blood pressures (P < 0.02 and P < 0.002 respectively) and total cholesterol levels (P < 0.004) than the control subjects. Calculation of Cp did not alter the observation of stiffer aortas in the stroke cohort (P < 0.0007). 4. In both stroke patient and control cohorts, as expected, inverse trends were observed between aortic compliance and blood pressure. Also as expected, in the control group Cp values did not show a relationship with blood pressure (r = 0.02, P = 0.092, not significant). However, in the stroke cohort a marked dependence of Cp on blood pressure was observed (r = -0.48, P = 0.03). 5. Transoesophageal echocardiographic studies have recently identified advanced atherosclerosis in the ascending aorta as a possible source of cerebral emboli and an independent risk factor for ischaemic stroke.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7493420 TI - Effects of maternal diabetes on blood pressure and glucose tolerance in offspring of spontaneously hypertensive rats: relation to birth weight. AB - 1. We studied the effects of maternal diabetes on blood pressure and glucose tolerance in the adult female offspring of spontaneously hypertensive rats. 2. Female spontaneously hypertensive rats were rendered diabetic by neonatal streptozotocin treatment, and then were mated with untreated male spontaneously hypertensive rats. Moderately severe hyperglycaemia was maintained during the gestation. 3. The birth weight was significantly lower in the female offspring of the diabetic dams than in the female offspring of the non-diabetic dams. The systolic blood pressure was significantly higher in the offspring from the diabetic dams than that from the control dams at 6 months of age (192 +/- 4 mmHg versus 213 +/- 4 mmHg, P < 0.01). The heart weight was also significantly increased in the offspring of the diabetic dams. Both the blood pressure and heart weight were inversely related to the birth weight. On the other hand, glucose tolerance was unaffected by maternal diabetes. 4. Maternal diabetes aggravated the severity of hypertension in the adult female offspring of spontaneously hypertensive rats. This suggests the importance of the metabolic environment during fetal growth for the development of hypertension. PMID- 7493421 TI - Circulating angiotensin-converting enzyme, von Willebrand factor antigen and thrombomodulin in exertional heat stroke. AB - 1. Military recruits frequently succumb to exertional heat stroke during intensive training. Since widespread endothelial injury is often associated with exertional heat stroke, the relationship between changes in three circulating endothelial cell markers (angiotensin-converting enzyme, von Willebrand factor antigen and thrombomodulin) and exertional heat stroke was studied. 2. Twelve recruits who had succumbed to exertional heat stroke during basic physical training (5000 m running) were included in the study. Another 10 age-matched healthy subjects who had gone through the same physical training regimen were selected as controls. 3. Blood was withdrawn on admission and on discharge for analyses of angiotensin-converting enzyme, von Willebrand factor antigen and thrombomodulin. Other physiological parameters and biochemical analyses reflecting renal and liver functions were also recorded. 4. Our results indicated that these subjects with exertional heat stroke exhibited impaired liver function as revealed by the significant elevation of both serum glutamic oxaloacetic transaminase (P < 0.05) and serum glutamic pyruvic transaminase (P < 0.05) as compared with normal healthy control subjects. Unfortunately, these values remained mostly somewhat elevated on discharge, although serum glutamic oxaloacetic transaminase was reduced dramatically. Indices of kidney functions, including creatinine clearance and uric acid and phosphorus secretion, were not significantly different from those observed in healthy controls. 5. Circulating angiotensin-converting enzyme activities in exertional heat stroke patients on admission were significantly lower than in normal subjects (10.68 +/- 2.15 versus 21.21 +/- 3.18 nmol hippuric acid min-1 ml-1, P < 0.05).(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7493422 TI - Effects of endopeptidase 24.11 inhibition on plasma and tissue concentrations of vasoactive intestinal peptide. AB - 1. In this study, we sought to determine the effect of endopeptidase 24.11 inhibition on the rate of metabolism of vasoactive intestinal peptide. The effect of such inhibition on the concentration of vasoactive intestinal peptide in two tissues was also investigated. 2. Male Sprague-Dawley rats were given the endopeptidase 24.11 blocker UK77,568 (10 mg/kg) or vehicle as a single intravenous injection or as a daily injection for 4 days. Two hours after the final or single injection, the rats were anaesthetized and blood was sampled to determine plasma concentrations of vasoactive intestinal peptide and angiotensin II. The hearts and kidneys were harvested and snap-frozen in liquid nitrogen. The plasma and tissue concentrations of vasoactive intestinal peptide and the plasma concentration of angiotensin II were determined by radioimmunoassay. In a separate group of experiments, male Sprague-Dawley rats were anaesthetized and carotid and jugular catheters were inserted. One hour after intravenous administration of UK77,568 or vehicle, an infusion of vasoactive intestinal peptide (10 pmol min-1 kg-1) was commenced via the jugular catheter. Blood was sampled to determine the vasoactive intestinal peptide concentration 1 h after commencing the vasoactive intestinal peptide infusion to calculate the metabolic clearance rate. 3. Plasma vasoactive intestinal peptide increased after acute (P < 0.05) but not chronic administration of UK77,568, while the concentration of vasoactive intestinal peptide in the heart increased after chronic administration (P < 0.0005). The concentration of vasoactive intestinal peptide in the kidney was unchanged after both acute and chronic endopeptidase 24.11 blockade. Plasma angiotensin II decreased significantly in the chronic group (P<0.05).(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7493423 TI - Effect of hypoxia and beta 2-agonists on the activity of the renin-angiotensin system in normal subjects. AB - 1. We have reported that the renin-angiotensin system is activated in acute asthma, and also by high-dose nebulized beta 2-agonists. The contribution of other possible stimuli such as hypoxia is unknown. The present study examined the effect of hypoxia alone and also combined with beta 2-agonists on the activity of the renin-angiotensin system. 2. In a double-blind crossover study, eight healthy subjects were randomized to inhale a hypoxic (FiO2 = 12%) or normoxic mixture for a period of 30 min, with either nebulized salbutamol (5 mg) or placebo administered into the circuit after 10 min. Plasma renin, angiotensin II and serum angiotensin-converting enzyme were measured at baseline and at intervals up to 2 h. Pulse rate and oxygen saturation were monitored continuously throughout the study. 3. After hypoxia alone, there was no change in the levels of plasma renin or angiotensin II. When salbutamol was added to the hypoxic mixture, there was a significant rise in plasma renin and angiotensin II [mean (SEM) maximal increase in angiotensin II of 5.6 (2.9) pg/ml and renin of 15.5 (6.3) mu-units/ml at 60 min, P < 0.05 compared with normoxia]. When salbutamol was administered in the normoxic mixture, plasma renin and angiotensin II also increased but this effect was similar to the effect of salbutamol in the hypoxic mixture. Serum angiotensin-converting enzyme levels were unaffected by hypoxia or salbutamol. 4. We conclude from these results that there is activation of the renin-angiotensin system in healthy subjects by salbutamol, but not hypoxia. (ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7493424 TI - Effect of oral digoxin, topical ouabain and salbutamol on transepithelial nasal potential difference in patients with cystic fibrosis. AB - 1. Airway epithelium in cystic fibrosis is characterized by a defect in chloride secretion across the apical membrane and an increase in sodium absorption. The increased rate of sodium absorption can be inhibited in vitro by ouabain, a Na(+) K(+)-ATPase inhibitor, and in cystic fibrosis patients the number and activity of nasal epithelial Na(+)-K(+)-ATPase pumps is increased. 2. We have performed a series of studies to determine whether drugs which modify airway epithelial Na(+) K(+)-ATPase activity in vitro can modify nasal potential in cystic fibrosis patients in vivo. As transepithelial nasal potential difference measurements were used to study the effect of drug modulation of airway epithelial ion transport in vivo, the repeatability of the technique was first evaluated. In order to assess the effectiveness of the technique used for measuring nasal potential difference, a pilot study was carried out using topical amiloride, a drug which has previously been shown to inhibit airway epithelium sodium transport in vivo. We then studied the effects of ouabain and digoxin, two inhibitors of Na(+)-K(+) ATPase, on nasal potential difference. 3. In study 1, nasal potential difference measurements were repeated on non-consecutive days in 20 patients with cystic fibrosis and 20 healthy individuals. Healthy subjects had a mean (SEM) potential difference value of -19.5 (0.9) mV with a 95% range for a single estimate of 75 133%. In patients with cystic fibrosis, the mean (SEM) potential difference was 40.4, (2.1) mV, with a 95% range for a single estimate of 74-136%.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7493425 TI - Isogravitational heterogeneity of perfusion after unilateral lung transplantation. AB - 1. Single-photon emission computerized tomography in both an intact canine model and man has demonstrated an aspect of pulmonary perfusion to be independent of gravitational forces. 2. Using technetium 99m-labelled macroaggregated albumin single-photon emission computerized tomographic imaging, we investigated normal human subjects (n = 5), stable unilateral lung transplant recipients (n = 6) and transplant recipients with chronic allograft dysfunction related to obliterative bronchiolitis (n = 5). 3. In coronal isogravitational sections, a 1 x 1 x N pixel strip (medial to lateral) was constructed through the 'core' pixel of maximal radioactive counts. The counts were measured for the 'core' pixel and at two mid points (medial and lateral) between the core pixel and the lung edges. Coefficients of variation were computed for each isogravitational strip and compared between groups. Fractional whole-lung perfusion was determined for left versus right lungs of normal subjects and allograft versus native lungs of transplant recipients. 4. Using these indices, 'isogravitational heterogeneity' (i.e. increased 'core' versus peripheral perfusion) was observed in allografts and native diseased lungs after unilateral transplantation. Despite significantly increased fractional whole-lung perfusion directed to the allografts (84.8% +/- 3.0% and 75.8% + 12.1% for stable unilateral lung transplant recipients and patients with obliterative bronchiolitis respectively) compared with normal lungs (50.2% +/- 1.2% and 49.8% +/- 1.2% for left and right respectively), 'isogravitational flow heterogeneity' (i.e. increased 'core' versus peripheral perfusion) was preserved after transplantation. 5. These findings suggest that 'isogravitational heterogeneity' was maintained despite increased unilateral pulmonary perfusion and the presumed increase in pulmonary capillary recruitment and/or distension. PMID- 7493426 TI - Effect of the transmembrane gradient of magnesium and sodium on the regulation of cytosolic free magnesium concentration in human platelets. AB - 1. To clarify the mechanism by which cytosolic free Mg2+ concentrations ([Mg2+]i) are regulated in human platelets, we investigated platelet membrane permeability to Mg2+ by altering extracellular Mg2+ concentrations, and tested the existence of a Na(+)-Mg2+ exchanger by manipulating the transmembrane Na+ gradient. 2. Platelet [Mg2+]i was 421 +/- 52 mumol/l in healthy men. [Mg2+]i remained constant during a 120 min exposure to nominally zero (low) or 5 mmol/l (high) external Mg2+ concentrations. 3. Preincubation of platelets with 10(-4) mol/l ouabain effectively decreased the transmembrane Na+ gradient. The ouabain-induced increase in [Mg2+]i was statistically significant after 30 min of exposure (14.6 +/- 2.0%) and reached 24.6 +/- 4.5% after 60 min. Similarly, the replacement of extracellular Na+ with N-methyl-D-glucamine significantly increased [Mg2+]i by 48.5 +/- 3.9% and 78.8 +/- 12.5%, respectively. 4. These results suggest that [Mg2+]i is well controlled in the presence of large transmembrane Mg2+ concentration gradients, and a Na(+)-Mg2+ exchanger may be involved in the regulation of [Mg2+]i in human platelets. PMID- 7493427 TI - Effect of water-loading on the performance of polyethylene glycol as a marker of small intestinal permeability. AB - 1. Polyethylene glycol has been used extensively to measure small intestinal permeability in vivo. However, polyethylene glycol seems to traverse the intestinal mucosa in much greater quantities than sugar molecules of equivalent M(r). In addition, the recovery of the lowest M(r) polymers of administered polyethylene glycol has been found to be both low and unreliable. 2. To compare the behaviour of a range of polyethylene glycol polymers with sugar probes in vivo, a combined polyethylene glycol/mannitol/lactulose probe was administered sequentially to healthy individuals in the fasted state and under conditions of water-loading. Timed hourly urine collections were made for 6 h. 3. Mannitol and lactulose recoveries were all within the normal range and were unaffected by co administration of water. The lactulose/mannitol recovery ratios did not vary significantly over the 6 h collection period. In contrast, the recovery of total polyethylene glycol was significantly greater when subjects were water-loaded. Furthermore, proportionally greater quantities of polyethylene glycol M(r) 370 than M(r) 854 were recovered towards the end of the collection period than at the start. 4. Our results show that, in contrast to lactulose and mannitol, excretion of low-medium M(r) polyethylene glycol polymers is highly dependent on co administration of water. Furthermore, the differential rate of excretion of the low compared with the high M(r) polyethylene glycol polymers suggests that the volume of distribution of the individual polymers may vary with M(r), and smaller polyethylene glycol molecules may undergo considerable renal tubular reabsorption. PMID- 7493428 TI - Effect of water restriction during adaptation to a low-sodium diet in man. AB - 1. Plasma sodium concentration may influence renal sodium excretion. We have examined the possibility that the fall in plasma sodium that occurs during salt restriction in man might be an important stimulus for renal sodium conservation. 2. In order to prevent the fall in plasma sodium that usually occurs during dietary salt restriction, we water restricted (200 ml/day) six normal subjects for the 2 days after the transition from 260 (high-sodium diet, day 3) to 20 mmol (low-sodium diet, days 4 and 5) sodium per day. In the control (hydrated) group water intake was held constant at 1800 ml/day. 3. Plasma sodium fell during the low-sodium diet in the hydrated group but remained constant in the dehydrated group (141.3 +/- 0.2 to 140.2 +/- 0.2 mmol/l versus 141.1 +/- 0.3 to 141.3 +/- 0.3 mmol/l). Plasma arginine vasopressin concentration was significantly higher and urine flow lower during the low-sodium diet in the dehydrated group (arginine vasopressin on day 5: hydrated group, 0.72 +/- 0.1 pmol/l; dehydrated group, 2.18 +/- 0.5 pmol/l). Weight fell by a similar amount in both groups (hydrated group, 1.23 +/- 0.17 kg; dehydrated group, 1.45 +/- 0.19 kg). 4. On the low-sodium diet there were no differences between groups in changes in plasma renin activity (hydrated group, 1.6 +/- 0.24 to 4.78 +/- 0.65 nmol angiotensin I h-1 ml-1; dehydrated group 1.57 +/- 0.18 to 5.14 +/- 0.56 nmol angiotensin I h-1 ml-1) or atrial natriuretic peptide (hydrated group, 23 +/- 2.3 to 14.7 +/- 1.6 pg/ml; dehydrated group, 26.8 +/- 3.6 to 12.7 +/- 1.3 pg/ml).(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7493430 TI - Beta-cell function in hypopituitary adults before and during growth hormone treatment. AB - 1. We studied beta-cell function in 40 hypopituitary adults and in 36 matched control subjects. Hypopituitary patients were studied again at 1, 3 and 6 months during a double-blind placebo-controlled trial of growth replacement lasting for 6 months. Biosynthetic human growth hormone was given subcutaneously in a daily dose of 0.02-0.05 i.u./kg at bed time. Fasting insulin, intact proinsulin and 32 33 split proinsulin were measured by two-site immunoradiometric assays. 2. Hypopituitary patients were aged 19-67 years and had a body mass index of 27.7 (18.0-41.1) kg/m2. They were receiving replacement thyroxine, adrenal steroids and sex hormones and they were growth hormone deficient. Control subjects were matched for age, sex and body mass index. Hypopituitary patients with normal glucose tolerance and with impaired glucose tolerance were compared separately with subgroups of control subjects matched for age and body mass index. 3. Twenty six hypopituitary patients had normal glucose tolerance and 14 had impaired glucose tolerance. All control subjects had normal glucose tolerance by World Health Organization criteria. Patients with impaired glucose tolerance were significantly older than those with normal glucose tolerance (P < 0.03). Hypopituitary patients with normal glucose tolerance compared with normal control subjects had a significantly lower fasting plasma glucose concentration (P < 0.01), a lower fasting insulin concentration (P < 0.006), a lower insulin-glucose ratio (P < 0.02) and a lower percentage of insulin to total insulin-like molecules [hypopituitary patients, 90% (81-96%); control subjects, 93% (78-97%); P < 0.02]. Hypopituitary patients with impaired glucose tolerance had similar glucose and insulin concentrations and insulin-glucose ratios as matched control subjects.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7493429 TI - Protein synthesis in isolated enterocytes from septic or endotoxaemic rats: regulation by glutamine. AB - 1. We studied the effect of sepsis and the regulation by glutamine of protein synthesis in enterocytes isolated from the small intestine of rats. 2. Sepsis was induced by caecal ligation and puncture; control rats were sham operated. Enterocytes were isolated from the jejunum and incubated in a medium containing [3H]phenylalanine. 3. Sixteen hours after caecal ligation and puncture, protein synthesis, measured as incorporation of radioactivity into protein, was increased by 65%, 89% and 137% respectively in enterocytes from the tips and mid-portions of the villi and from the crypts. 4. Addition of glutamine to incubated enterocytes stimulated protein synthesis in a dose-dependent manner, and this effect was most pronounced in crypt cells from septic rats. The effect of glutamine on protein synthesis was duplicated by equimolar concentrations of acetoacetate or 3-hydroxybutyrate, both of which may serve as fuel for enterocytes, and was blocked by the glutaminase inhibitor 6-diazo-5-oxo-L norleucine. 5. The results suggest that sepsis stimulates protein synthesis in enterocytes and that glutamine regulates protein synthesis in the same cells, probably by energy provision. PMID- 7493431 TI - Total antioxidant capacity and lipid peroxidation during liver transplantation. AB - 1. We assessed plasma total antioxidant capacity using enhanced chemiluminescence and plasma lipid peroxides in eight patients undergoing orthotopic liver transplantation and 26 healthy control subjects. 2. The antioxidant capacity of plasma decreased significantly on reperfusion of the donor liver (P < 0.05) during liver transplantation, although levels were not lower in patients than in control subjects. 3. Pre-reperfusion antioxidant capacity correlated negatively with the concentration of lipid peroxidation products appearing in the plasma by the end of the transplant (P < 0.02), but did not correlate with vitamin A or E concentrations. 4. This study provides evidence of oxidant stress during liver transplantation, as shown by antioxidant utilization associated with increased lipid peroxidation. PMID- 7493432 TI - Smoking and plasma lipoprotein metabolism. PMID- 7493433 TI - Lack of effect of vitamin E administration on basal nitric oxide function in male smokers and non-smokers. AB - 1. The antioxidant properties of vitamin E (alpha-tocopherol) have been proposed to account for a possible protective effect in coronary artery disease. Coronary risk factors are associated with depressed endothelium-derived nitric oxide responses, and some evidence suggests increased quenching of the vasodilator effect of nitric oxide by oxygen free radicals. This study examined the possibility that vitamin E supplementation would improve basal function of the nitric oxide dilator system in long-term smokers and non-smokers. 2. Nine healthy male life-long non-smoking subjects and eight smokers (36 +/- 6 cigarettes per day, > 7 years) were given 1000 i.u. of vitamin E per day for a 4-week period, while five additional non-smokers received placebo. Forearm blood flow responses to intra-arterial infusion of noradrenaline (60, 120 and 240 rho mol/min) and the competitive inhibitor of nitric oxide synthesis, NG-monomethyl-L-arginine (1, 2 and 4 mumol/min) were measured before and after therapy. 3. No changes were evident in the constrictor responses to noradrenaline or NG-monomethyl-L-arginine in any group. These results indicate that vitamin E supplementation does not influence basal activity of the nitric oxide-dilator system in vivo. PMID- 7493434 TI - Postprandial sympatho-adrenal activity: its relation to metabolic and cardiovascular events and to changes in meal frequency. AB - 1. Sympatho-adrenal activity was measured after the consumption of a 3.15 MJ mixed meal. Whole-body noradrenaline spillover rates, forearm plasma noradrenaline spillover and adrenaline secretion rates were derived using isotope dilution methodology. Heart rate and blood pressure spectral analysis measurements were also made. The relation of sympathoadrenal activity to thermogenic and cardiovascular events was studied. Sympathetic nervous and thermogenic responses were measured for 120 min after the single 3.15 MJ meal and compared with those after three 1.05 MJ meals, given 30 min apart. 2. Whole-body and forearm plasma noradrenaline spillover, and the 0.1 Hz component of systolic pressure power all increased significantly postprandially, while the 0.1 Hz component of heart rate variability, an indirect index of cardiac sympathetic nervous activity, remained unaltered. Adrenaline secretion was unaltered postprandially. Whole-body plasma noradrenaline spillover and thermogenesis during the 120 min postprandial period were 37% and 36% higher after the single meal as compared with the multiple meals, although this was not statistically significant. 3. The sympathetic neural responses were delayed in relation to peak plasma insulin levels and sustained in the face of declining insulin levels. Energy expenditure increased significantly postprandially, but there was no direct quantitative relationship to plasma noradrenaline spillover. Forearm oxygen consumption did not increase postprandially despite significant increases in regional noradrenaline spillover. Thus, no close relation was demonstrated between postprandial sympathetic nervous activation and either insulin secretion or thermogenesis. PMID- 7493436 TI - Effect of the somatostatin analogue, octreotide, on exercise-induced hypotension in human subjects with chronic sympathetic failure. AB - 1. In autonomic failure, supine exercise lowers blood pressure and worsens postural hypotension. The somatostatin analogue, octreotide, reduces post prandial and postural hypotension, but its effects on exercise-induced hypotension and on postural hypotension post-exercise are unknown. 2. Eighteen subjects with chronic sympathetic denervation were studied; 12 had pure autonomic failure and six had additional neurological features of the Shy-Drager syndrome. Haemodynamic, hormonal and biochemical changes were measured before, during and after incremental supine leg exercise on two occasions: on no treatment and after subcutaneous octreotide. Exercise was performed 120 min after octreotide in eight subjects and 60 min after octreotide in ten subjects. 3. Octreotide did not improve exercise-induced hypotension; the blood pressure fall was greater during exercise, but the blood pressure level was no different than without treatment. Heart rate, stroke distance, cardiac index and systemic vascular resistance were similar at rest and changed to the same degree with exercise on and off octreotide. After octreotide, resting levels of serum growth hormone, plasma noradrenaline, adrenaline and renin were unchanged, but glucose was higher and insulin was lower. There was no change in biochemical and hormone levels during exercise either off or on octreotide. 4. After octreotide, although the rate of blood pressure recovery was similar post-exercise, the levels of blood pressure were higher than in the non-treatment phase and postural hypotension was improved before and after exercise. 5. In conclusion, in primary autonomic failure, octreotide did not improve exercise-induced hypotension in the supine position, suggesting that octreotide-sensitive vasodilatory peptides do not contribute to the blood pressure fall.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7493435 TI - Antecedent hypoglycaemia in non-diabetic subjects reduces the adrenaline response for 6 days but does not affect the catecholamine response to other stimuli. AB - 1. Recent hypoglycaemia reduces the counter-regulatory and symptomatic response to the next episode of hypoglycaemia. This study was designed to determine whether antecedent hypoglycaemia (a) affected the sympathoadrenal response to other stimuli (postural change and a mathematical stress test, (b) was associated with adaptation of cognitive function and (c) if avoided for 6 days led to the restoration of the counter-regulatory response to hypoglycaemia. 2. On the morning of day 1, 10 healthy men had their response to the sympathoadrenal stimuli assessed before an episode of hypoglycaemia (2.5 mmol/l). A further hyperinsulinaemia clamp was performed for 2h (blood glucose 3.0 mmol/l or 5.0 mmol/l) in the afternoon and the assessments repeated the next day and 6 days later. Cognitive function using the four-choice reaction time test was assessed before and during the episodes of morning hypoglycaemia. The whole study was repeated 4 weeks later with the other afternoon glucose level. 3. Adrenaline levels and sweat rates were reduced (P < 0.05) on day 2 during hypoglycaemia on both occasions and had not returned to normal after 6 days. Symptom scores were lower (P < 0.05) only on day 2 after the afternoon of hypoglycaemia. The responses to the sympathoadrenal stimuli and cognitive function were unaffected by antecedent hypoglycaemia. 4. The adaptation of the counter-regulatory response to antecedent hypoglycaemia appears to be specific to hypoglycaemia. The response takes longer than 6 days but less than 4 weeks to return to normal. Antecedent hypoglycaemia does not lead to cerebral adaptation. PMID- 7493437 TI - Cholecystokinin is a satiety hormone in humans at physiological post-prandial plasma concentrations. AB - 1. Intravenous infusions of the brain/gut hormone, cholecystokinin, have been shown to reduce food intake in a subsequent test meal. However, in previous studies the doses administered were large and likely to have produced plasma concentrations far in excess of the normal post-prandial range. 2. In this study cholecystokinin-8 was infused intravenously to six healthy subjects in doses that reproduced physiological post-prandial concentrations. Plasma concentrations of cholecystokinin were measured using a novel sensitive and specific radioimmunoassay. The effect of cholecystokinin-8 infusion on subsequent food intake in a standard test meal was compared with the effect of saline infusion in the same subjects. 3. Food intake (mean +/- SEM) was significantly less during cholecystokinin (5092 +/- 665 kJ) than during saline infusion (6418 +/- 723 kJ, P = 0.03). During cholecystokinin infusion, plasma concentrations increased from 0.45 +/- 0.06 pmol/l to 7.28 +/- 2.43 pmol/l immediately before the meal. With saline infusion there was no premeal increase in plasma cholecystokinin concentration. 4. This paper describes a novel radioimmunoassay for measurement of plasma concentrations of cholecystokinin. Using this assay we have demonstrated that cholecystokinin is important in control of satiety in humans. PMID- 7493438 TI - Effect of dietary restriction on protein synthesis and wound healing after surgery in the rat. AB - 1. The healing of an abdominal muscle wound after surgery is associated with a considerable increase in the rate of protein synthesis. We have investigated whether this increase in protein synthesis is affected by chronic undernutrition, and whether this causes a delay in wound healing. 2. A group of rats was fed 58% of the voluntary food intake of a matched control group. After 7 days half the rats in each group underwent abdominal surgery. Forty-eight hours later all the rats were killed and muscle protein synthesis rate was measured by the flooding dose technique. 3. In a second experiment using the same dietary regimen rats were placed in metabolic cages after surgery and killed 7 days later. In addition to measurements of muscle protein synthesis, wound breaking strength was measured with a tensiometer and collagen content was also measured at the wound site. 4. Dietary restriction caused a loss of body weight, a decrease in nitrogen balance and a deficit in muscle protein mass. It also caused a decrease in protein synthesis rate in gastrocnemius muscle and in parts of the abdominal muscle distant from the site of the wound. However, it had no effect on the rate of muscle protein synthesis at the site of the wound either 2 or 7 days after surgery. The tensile strength and the collagen content of the wound were also unaffected by food restriction. 5. It is concluded that the wound healing process is uniquely protected from the effects of moderate undernutrition such as might be experienced by a chronically ill patient. PMID- 7493439 TI - Whole-body protein turnover and hepatic protein synthesis are increased by vaccination in man. AB - 1. The ability of diphtheria-tetanus-poliomyelitis-typhoid vaccination to induce modifications in protein metabolism was investigated in post-absorptive healthy humans. 2. Seven subjects were studied before and 2 days after vaccination. They underwent an intravenous primed constant infusion of L-[1-13C]leucine for 4h. Plasma protein concentrations, whole-body amino acid fluxes and acute-phase protein synthesis were determined. 3. Plasma concentrations of fibrinogen, alpha 1-acid glycoprotein, haptoglobin and alpha 1-antitrypsin were significantly elevated 2 days after vaccination (P < 0.05). Leucine oxidation was unaffected but whole-body protein synthesis and breakdown were both increased (P < 0.05), by 25 and 16% respectively, in subjects who had an elevated body temperature (n = 5). Albumin synthesis was unchanged, but hepatic synthesis of fibrinogen was 56% higher after vaccination. 4. The present investigation indicates that diphtheria tetanus-poliomyelitis-typhoid vaccination could induce a sustained acute-phase reaction. Moreover, protein metabolism appeared to be extremely sensitive to a mild stress since leucine kinetics and fibrinogen synthesis were affected. Therefore, diphtheria-tetanus-poliomyelitis-typhoid vaccination might represent an attractive model for studying the inflammatory process in humans. PMID- 7493441 TI - Influence of blood group and secretor status on carbohydrate structures in human gastric mucins: implications for peptic ulcer. AB - 1. The content and distribution of carbohydrate was examined in mucus glycopolypeptides from human antral mucosae. 2. The mean amount of carbohydrate per 1000 amino acid residues was found to be similar in glycopolypeptides with A, B or H activity. It was slightly, though significantly, less in glycopolypeptides lacking these determinants, because carbohydrate chains were of a shorter average length than in the A-, B- or H-active preparations. This difference was reflected in the sizes of oligosaccharide-alcohols released from representative glycopolypeptides with alkaline borohydride. 3. Differences between A-, B- or H active and non-secretor glycopolypeptides in terms of the mean number of carbohydrate chains per 1000 amino acid residues were found to be small, and without significance. 4. The average number of peripheral monosaccharide units per 1000 amino acid residues was greater in A-active than in H-active, and least in non-secretor, glycopolypeptides. This order was reversed for monosaccharide units incorporated into skeletal (core plus backbone) structures. The difference in each case was statistically significant. 5. These findings suggest that the increased risk of peptic ulcer associated with blood group O and non-secretor status is unlikely to be attributable to an inherent deficiency in the protective mucus layer, linked to differences between mucins that are associated with A, B or H activity. Other hypotheses linked to infection with Helicobacter pylori are examined. PMID- 7493440 TI - Expression and regulation of glycosyltransferases for N-glycosyl oligosaccharides in fresh human surgical and murine tissues and cultured cell lines. AB - 1. Mammalian membrane and serum proteins are glycosylated by the addition of heterogeneous N-linked oligosaccharides. It has been widely speculated that oligosaccharide diversity is achieved by corresponding heterogeneity of expression of the glycosyltransferases that are responsible for oligosaccharide synthesis. 2. We surveyed mRNA levels of three sequentially acting glycosyltransferases, N-acetylglucosaminyl-transferase I, beta 1,4 galactosyltransferase and alpha 2,6-sialyltransferase, in 11 human tissues and confirmed the expected variations. 3. The size heterogeneity of alpha 2,6 sialytransferase transcripts reported in rat tissues was evident neither in the human tissue survey nor in a panel of murine RNAs. Tissue distributions of alternative terminal sialyltransferases, alpha 2,6-sialyltransferase and alpha 2,3-sialyltransferase, were distinct. 4. Relative glycosyltransferase mRNA levels in four transformed human cell lines cultured in vitro did not fully reflect levels in the corresponding human tissues. 5. Expression of alpha 2,6 sialyltransferase mRNA was approximately 2.6-fold greater in adenocarcinomatous than in normal human colon, and beta 1,4-galactosyltransferase expression was approximately 1.8-fold greater in normal than in adenocarcinomatous colon. 6. n Butyrate (0.003-0.005 mol/l), a short-chain fatty acid that is produced by colonic bacterial fermentation, caused approximately 80% inhibition of alpha 2,6 sialyltransferase, approximately 2.5-fold induction of beta 1,4 galactosyltransferase and approximately 6-fold induction of N acetylglucosaminyltransferase mRNAs in T84 (colonic) cells. The effects on alpha 2,6-sialyltransferase and beta 1,4-galactosyltransferase were near maximal by 6h, but induction of N-acetylglucosaminyltransferase was fully apparent only after exposure for 24 h. PMID- 7493442 TI - High-affinity interaction of long-chain fatty acids with serum albumin in nephrotic syndrome. AB - 1. We have examined the effect of hypoalbuminaemia, a hallmark of nephrotic syndrome, on the albumin-fatty acid equilibrium in the plasma of 11 adult patients with nephrosis compared with 12 healthy subjects and six subjects with normoalbuminaemic hyperlipoproteinaemia. 2. We used a dialysis exchange rate method which allows the evaluation in relative terms of the binding affinity of albumin for plasma fatty acid and the fatty acid availability, tentatively equated with the unbound fatty acid fraction. 3. In nephrotic patients, an increase (P < 0.001) in albumin affinity for fatty acid was seen compared with healthy subjects, which was negatively correlated with albuminaemia (r = -0.69, P < 0.02). No change in fatty acid availability was seen for the group as a whole, but individual values showed a wide scatter, with the highest values in four patients with the highest fatty acid-albumin molar ratios. The increase in albumin affinity for fatty acid was specific to nephrotic syndrome since no such effect was seen in subjects with hyperlipoproteinaemia, who only showed a moderate increase (P < 0.01) in fatty acid availability. 4. The increased albumin affinity for fatty acid in nephrotic syndrome supports the hypothesis that an albumin component with lower affinity for fatty acid might filter out through the diseased glomerular membrane and leave the high-affinity albumin in plasma. PMID- 7493443 TI - Effects of weight reduction on the regulation of lipolysis in adipocytes of women with upper-body obesity. AB - 1. The regulation of lipolysis was studied in 14 upper-body obese women aged 26 55 years. Isolated subcutaneous adipocytes from the abdominal region were examined before and after an 8 to 12-week-long weight reduction programme, during which the mean body mass index (kg/m2) of the group was reduced from 36.4 +/- 0.9 to 30.3 +/- 1.0. 2. Fat cell volume was reduced from 891 +/- 39 to 655 +/- 45 pl. The sensitivity to noradrenaline stimulation of lipolysis in vitro increased fivefold after weight reduction. A corresponding increase in sensitivity was found with the beta 2-selective adrenoceptor agonist terbutaline. However, the number of beta 2-adrenoceptor binding sites as assessed by radioligand binding with 125I-labelled cyanopindolol was not changed. No changes were observed when dobutamine (a beta 1-selective adrenoceptor agonist) and clonidine (an alpha 2 adrenoceptor agonist) were used. 3. The basal lipolysis rates decreased by about 50% after weight reduction and the maximum enzyme activity of hormone-sensitive lipase was also reduced by almost 50%. 4. Plasma concentrations of insulin, noradrenaline and total testosterone decreased and sex hormone-binding globulin increased after weight reduction. Calculated apparent free testosterone levels decreased by more than 40% after weight reduction. 5. In conclusion, weight reduction leads to increased efficiency of adipocyte lipolysis with decreased resting lipolysis rate but increased sensitivity to stimulation by catecholamines, which may be attributed to a decreased activity of hormone sensitive lipase and an increased sensitivity of beta 2-adrenoceptors. Changes in circulating levels of catecholamines, insulin and testosterone may play a role in these modifications of adipocyte function.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7493444 TI - Interleukin-6 induces proteolysis by activating intracellular proteases (cathepsins B and L, proteasome) in C2C12 myotubes. AB - 1. A cell culture system of C2C12 myotubes was established as a model of the muscle. With the aid of this model, the half-lives of intracellular proteins as well as the activities and mRNA levels of proteasomes (26S and 20S) and cathepsins (B, L, and H) were examined in the presence of various amounts of cytokines. 2. It was found that 100 units/ml recombinant human interleukin-6 somewhat shortened the half-life of long-lived proteins to 23.79 +/- 1.55 h (control: 25.60 +/- 1.87 h). When 1% fetal bovine serum contained in the culture medium was replaced by 0.5 mg/ml bovine serum albumin, interleukin-6 was more effective since 10 units/ml of interleukin-6 shortened the half-life to 19.09 +/- 2.87 h (control: 22.26 +/- 321 h). Interleukin-6 (100 units/ml) increased the activity of 26S proteasome by 31.5%, of cathepsin B by 53.5% and of cathepsin B+L by 21.3%. These increases occurred in association with an increase in their transcription. 3. On the other hand, 1000 units/ml of recombinant human tumour necrosis factor alpha prolonged the half-life of long-lived proteins while reducing the protease activities of 20S proteasome (-27.1%), cathepsins B ( 64.6%) and B+L (-54.9%). 4. These results suggest that interleukin-6 induces degradation of long-lived intracellular proteins by activating both the non lysosomal (proteasomes) and lysosomal (cathepsins) proteolytic pathways. It is therefore concluded that interleukin-6 is a candidate for a proteolysis-inducing factor in myotubes and may play an important role in the progression of muscle degradation in systemic inflammatory responses induced by sepsis or severe injury. PMID- 7493445 TI - Effect of breathing, pressure and posture on palatoglossal and genioglossal tone. AB - 1. Patency of the upper airway is critical to respiration. Although about half of patients with the sleep apnoea/hypopnoea syndrome obstruct their upper airway at the retropalatal level, the respiratory actions of the palatal muscles have been little studied. We have therefore tested the hypothesis that the nasopharyngeal dilator muscle palatoglossus is activated during inspiration and by negative pressure. 2. Using intramuscular wire electrodes inserted perorally, we have compared the response of palatoglossus and genioglossus to breathing, posture change and airway negative pressure in 10 normal awake subjects before and after topical anaesthesia. The results are expressed as a percentage of maximal electromyogram. Data were analysed by repeated-measures analysis of variance. 3. Inspiratory activity was exhibited by both genioglossus [inspiratory, 10% +/- 2% (SEM); expiratory, 6% +/- 1%, P = 0.001] and palatoglossus (inspiratory, 16% +/- 5%, expiratory, 10% +/- 3%, P = 0.016), but only genioglossus exhibited increased activity on lying (supine 10% +/- 2%, erect 6% +/- 1% maximum, P = 0.01). 4. One hundred milliseconds after negative pressure application, activity increased in both genioglossus (7% +/- 2% and 13% +/- 3% respectively, P = 0.02) and palatoglossus (8% +/- 2% and 23% +/- 6% respectively, P < 0.001). After lignocaine surface anaesthesia to the nose and pharynx both genioglossus and palatoglossus still increased their activity in response to negative upper airway pressure, the extent of the increase being decreased for palatoglossus (P = 0.02) but not for genioglossus. 5. Thus, palatoglossus has respiratory activity and is activated by negative upper airway pressure. PMID- 7493446 TI - Effects of treadmill exercise and high-fat feeding on muscle degeneration in mdx mice at the time of weaning. AB - 1. Dystrophin-deficient hindlimb muscles of mdx mice undergo necrosis at the time of weaning when the motor activity of the mice greatly increases and muscle energy metabolism becomes more dependent on insulin and carbohydrates. 2. We have attempted to determine if the onset of myofibre necrosis in mdx mice at the time of weaning is related to the development of motor activity and/or the change in diet. 3. Fourteen-day-old mdx mice were divided into two groups after weaning. One group was trained to run on a treadmill and the other group was kept on a high-fat diet. Muscle necrosis was assessed histologically in the soleus and extensor digitorum longus muscles of mice in both experiments. 4. Keeping mice on a high-fat milk diet from the time of weaning up to 42 days of age did not influence the occurrence of necrosis in the soleus and extensor digitorum longus muscles of the mdx pups. In contrast, treadmill exercise greatly increased necrosis in both muscles. 5. We conclude that an increase in motor activity exacerbates the degeneration of hindlimb muscles of mdx mice at the time of weaning. PMID- 7493447 TI - Systemic treatment with epidermal growth factor induces anaemia in Goettingen minipigs. AB - 1. From two independent experimental studies conducted in 48 minipigs with oesophageal sclerotherapy and concomitant treatment with epidermal growth factor, blood samples together with bone marrow biopsies were analysed for safety data. 2. Four to five weeks of systemic treatment with epidermal growth factor induced a decline in blood haemoglobin concentration in a time- and dose-dependent and reversible manner but without an effect on leucocyte or platelet counts. 3. The bone marrow expressed decreased amounts of haematopoietic tissue and reduced numbers of erythropoietic cells. 4. Four to five weeks of systemic treatment with epidermal growth factor induced reversible increases in serum concentrations of creatinine and urea, most likely reflecting renal impairment. 5. Groups in which creatinine and urea were not increased also had reduced blood haemoglobin concentrations. 6. These findings suggest that epidermal growth factor selectively impaired the erythropoiesis and stress the importance of risk-benefit analysis concerning the potential therapeutic applications of epidermal growth factor. PMID- 7493449 TI - Vth International Symposium on Sjogren's Syndrome. Noordwijkerhout, The Netherlands, June 15-17, 1995. Proceedings and abstracts. PMID- 7493448 TI - Endothelium-mediated vascular dysfunction in diabetes. PMID- 7493450 TI - Ro/SS-A and La/SS-B: autoantigens in Sjogren's syndrome? AB - Sjogren's syndrome is characterized serologically by the presence of autoantibodies against Ro/SS-A and La/SS-B. The mechanisms by which these autoantibodies arise is not clear at this moment. B cells capable of producing antibodies to Ro/SS-A and La/SS-B seem to be present in every individual; whether or not an individual indeed makes these antibodies is governed by T cells. Recent experiments from us and from others indicate that T cell tolerance towards Ro/SS A and La/SS-B can be broken by immunization of normal mice with recombinant human Ro/SS-A or La/SS-B. The specificity of the T cells directing the anti-Ro/SS-A and anti-La/SS-B autoantibody response in these animals has not yet been elucidated. T cells may either be directed against foreign epitopes present on the human immunogen or they may be truly autoreactive. In patients with Sjogren's syndrome, a comparable immunization route might encompass product of viral origin directing the T cell response via RNA/protein complexes. Putative candidates comprise viruses that make use of RNAse polymerase-III, such as Epstein Barr virus, adenovirus, vesicular stomatitis virus en rabies virus. PMID- 7493451 TI - Vth International Symposium on Sjogren's syndrome. Clinical aspects and therapy. AB - There is increasing awareness that sicca syndrome can lead to ocular, oral and systemic complications. However, a wide variety of pathogenetic processes may lead to the same end result of diminished glandular secretion. Recognition of these different etiologies will lead to improved therapies. On the one hand, an active inflammatory/autoimmune process may respond to immunomodulatory drugs. On the other hands, processes that predominantly involve "neuroendocrine" dysfunction (unrelated to immune activity) of the gland will require a different approach. PMID- 7493452 TI - Classification of disease manifestations in primary Sjogren's syndrome: present status and a new proposal. AB - Establishing a model for classification of the clinical disease manifestations in primary Sjogren's syndrome is a challenge with important implications for handling individual patients and for describing and analyzing patient materials. Based on the pathobiology of primary SS we define three (1-3) "exocrine" and four (4-7) "nonexocrine" subgroups of disease manifestations. Accordingly, 1) "surface exocrine disease" includes the diagnostic features from eyes (keratoconjunctivitis sicca) and mouth (xerostomia), and the manifestations from upper airways (rhinitis sicca, xerotracheitis) and skin (xeroderma). Involvement of the excretory parenchyma of the lungs, hepatobiliary system, pancreas, intestinal tract and kidneys is designated 2) "internal organ exocrine disease". These manifestations are potentially severe, do not lead to subjective dryness, and none of them are diagnostic for the disease. We suggest 3) "monoclonal B lymphocyte disease" (lymphoma) to be an exocrine disease manifestation because it originates mostly from the immunoinflammatory foci of the autoimmune exocrinopathy. The nonexocrine manifestations are subgrouped into: 4) "inflammatory vascular disease" (vasculitis and perivasculitis), 5) "noninflammatory vascular disease" (Raynaud), 6) "mediator-induced disease" (hematologic cytopenia, fever and fatigue) and 7) "autoimmune endocrine disease". Subdividing the seven subgroups leads to a third order of classification in which single and separate manifestations are placed. The descriptive and analytic power of the proposed model for classification of disease manifestations in primary Sjogren's syndrome should be evaluated in clinical studies. PMID- 7493453 TI - Sjogren's syndrome--a house of cards. AB - Sjogren's syndrome is an autoimmune rheumatic disease that is the result of the interplay between a number of environmental and genetic factors. In this short review an analogy is suggested with a game of cards in which each card is marked with a particular feature. A particular set of cards when dealt together could constitute Sjogren's syndrome if the hand includes cards marked for female sex, HLA DRS, anti Ro antibodies, anti La antibodies, exposure to retroviruses etc. PMID- 7493454 TI - The sodium lauryl sulfate model: an overview. AB - Irritant contact dermatitis is a complex entity with several clinical forms (acute, cumulative, etc.) and multiple mechanisms. Sodium lauryl sulfate (SLS), the most widely utilized model for studying acute and cumulative irritation, has proved highly practical and informative for such studies. This article summarizes several decades of investigation, and provides details of dosing, application method (closed versus open), and biologic endpoints (visual grading, transepidermal water loss) that may be utilized in future studies. PMID- 7493455 TI - Structure-activity relationships in allergic contact dermatitis (I). Studies on the influence of side-chain length with derivatives of primin. AB - Primin (2-methoxy-6-pentyl-1,4-benzoquinone) is a naturally-occurring strong sensitizer from Primula obconica (Primulacease). To determine the effect of side chain length on sensitizing potency, 15 analogues with linear side chains from C1 to C15 and 4 C6-analogues with branched side chains were prepared synthetically and devoted to experimental sensitization in guinea pigs. The results showed an increase of the sensitizing capacity with increasing length of the alkyl side chain from C1 to C10, reaching a maximum at C11 and C12. On further elongation, the sensitizing potency decreased beyond C13, reaching values which finally were as low as those of the C1 and C3 derivatives. The results mirror findings which formerly have been obtained with other non-quinonoid compounds like catechols, phenols, hydroquinones and gallates. In the plant kingdom, compounds approximating an "ideal allergen" consisting of a quinonoid ring with a 10 or 11 carbon-membered side chain have been identified only once: 2,3-dimethoxy-geranyl 1,4-benzoquinone, a remarkably strong sensitizer found in Wigandia caracasana (Hydrophyllaceae). PMID- 7493456 TI - Contact allergy to allyl glycidyl ether present as an impurity in 3 glycidyloxypropyltrimethoxysilane, a fixing additive in silicone and polyurethane resins. AB - We present a 43-year-old man who worked in the plastics industry and who suffered from an occupational dermatitis on the hands and forearms. He was found to be allergic to an epoxy silane compound that was contaminated with allyl glycidyl ether, a reactive epoxy diluent. HPLC analysis and patch testing indicated that this impurity was probably the only sensitizer. Moreover, the gloves used provided absolutely no protection, as the inner side also generated strong positive reactions. PMID- 7493457 TI - Nickel/magnesium interactions in nickel-sensitive patients. AB - Experimentally demonstrated interactions between Ni2+ and Mg2+ were examined in human beings. 110 subjects patch-test-positive to 10 microliters aq. NiSO4 0.1 M were subdivided into groups of 30, 50 and 30 people. Each subgroup was tested to 10 microliters NiSO4 0.1 M solution as a control and to mixed solutions containing NiSO4 0.1 M together with, respectively MgSO4 0.1 and 0.3, 0.3 and 0.5 and 0.5 and 1 M. On increasing the applied concentrations of MgSO4, the % of patients with reduced or suppressed nickel reactions, with 1 exception, proportionally increased. The exception concerned testing with 0.5 M, where a paradoxically exacerbating increase in nickel reactions was seen in a majority of nickel-sensitive subjects. MgCl2 aq. at 0.3, 0.5 and 1 M concentrations was not able to reduce the cutaneous patch test positive reactions to NiCl2 0.1 M in 25 sensitive patients. On increasing the applied concentrations of MgCl2, both the number and intensity of patch test reactions to NiCl2 proportionally increased. A supposed role of the sulfate and chloride counterions in the penetration of nickel was examined in 30 NiSO4 5% patch-test-positive patients, testing to 10 microliters of aq. NiSO4 0.1 M, NiCl2 0.1 M, NiSO4 0.1 M + MgCl2 0.3 M, NiCl2 0.1 M + MgSO4 0.3 M, Na2SO4 0.3 M, NaCl 0.3 M, NiSO4 0.1 M + Na2SO4 0.3 M, NiCl2 0.1 M + NaCl 0.3 M. The findings suggest that the addition of sulfate or chloride to nickel could determine the formation of different Ni complexes directed toward different targets, one Mg(2+)-dependent, the other Mg(2+)-independent. PMID- 7493458 TI - An evaluation of the suitability of benzocaine as a positive control skin sensitizer. AB - Although they are subject to some limitations, sensitization tests such as the guinea pig maximization test (GPMT) have for many years provided a valuable basis for the identification of skin sensitization potential. Thus they have been used widely by regulatory authorities, such as those in Europe, as a means to identify significant sensitization hazards associated with new chemicals. However, the standard of performance of guinea pig sensitization assays has been demonstrated to be widely variable. Consequently, the OECD sensitization test guideline (the de facto world standard) has been updated to incorporate recommendations for action whose aim is to achieve a minimum standard of test conduct. The principle is that a test laboratory should be able to demonstrate an acceptable level of response using a moderately sensitizing chemical. A list of 3 such chemicals is provided, hexyl cinnamic aldehyde, mercaptobenzothiazole and benzocaine. It is our experience that whilst good results can readily be obtained with the first 2 of these, benzocaine is much more difficult. Using both the GPMT and the local lymph node assay (LLNA), an OECD-recommended screening test, benzocaine has given highly variable results. A range of from 0% to 60% positive in the GPMT was found and, in most tests, benzocaine would not classify as a skin sensitizer according to EU criteria. In the LLNA, from a series of 12 tests conducted in 2 laboratories, only occasional positive results were obtained. Furthermore, these positive results were not reproducible. Reasons for this variability are discussed. However, the main conclusion must be that benzocaine does not represent a useful moderately sensitizing positive control. PMID- 7493459 TI - Butenylbithiophene, alpha-terthienyl and hydroxytremetone as contact allergens in cultivars of marigold (Tagetes sp.). AB - Ornamental cultivars of Tagetes sp., commonly named marigold, are one of the presently most popular pot and garden plants. Sensitizing experiments in guinea pigs with short ether extracts and isolated compounds revealed the presence of 3 constituents that must be considered as contact allergens. They were identified as 5-(3-buten-1-ynyl)-2,2'-bithiophene, alpha-terthienyl and hydroxytremetone. In sensitized animals, butenylbithiophene showed moderate to strong sensitizing potency, while alpha-terthienyl was less strong and hydroxytremetone weak. The results demonstrate, for the first time, that at least some of the thiophenes abundantly occurring in many species of the Compositae family possess not only phototoxic activity but also sensitizing properties. PMID- 7493460 TI - Comparison of in vitro predictive tests for irritation induced by anionic surfactants. AB - Skin compatibility of anionic surfactants may often but not always be predicted by in vitro tests. For instance, the correlation between in vivo and in vitro data is classically hampered in the presence of magnesium. This ion is known to interfere with in vitro skin irritation predictive tests based on protein denaturation. This study was conducted to compare a recently introduced assay, corneosurfametry, with other in vitro tests including the pH-rise of bovine serum albumin, collagen swelling, and zein solubilization tests. Corneosurfametry entails collection of cyanoacrylate skin surface strippings and short contact time with surfactants, followed by staining samples with toluidine blue and basic fuchsin dyes. Measurements are made by reflectance colorimetry. Data show that irritation potentials predicted by corneosurfametry agree with those obtained by established in vivo and in vitro irritation tests. Moreover, corneosurfametry data are not artificially lowered by addition of magnesium in surfactant solutions. In conclusion, corneosurfametry should be viewed as one of the realistic predictive tests for surfactant irritancy. PMID- 7493461 TI - Skin problems among Danish dental technicians. A cross-sectional study. AB - Our objective was to evaluate changes in the prevalence of skin problems on the hands among dental technicians during the last 10 years by comparison with results of a previous study, to study whether the presence and location of skin problems could be related to work with acrylates, and to study whether the use of gloves had any influence on the prevalence of the skin problems. A cross sectional questionnaire study among 192 dental technicians at work was performed using an expanded version of a questionnaire from a Danish survey 10 years ago. The cumulative prevalence of skin problems on the hands was 53%. The 1-year prevalence of skin problems on the hands was 43%. The point prevalence of skin problems on the hands was 36%. These prevalence figures were profoundly higher than the figures reported for the general population, and the figures did not differ from those obtained in the study 10 years ago. The point prevalence of skin problems on the hands among individuals handling acrylates was 38% higher than among individuals who never worked with acrylates. No difference in the occurrence of skin problems was observed between individuals using gloves and individuals who did not use gloves while handling acrylates. The prevalence difference of skin problems on the fingertips was 7.3% between individuals handling acrylates daily or several times a week and individuals handling acrylates less frequently.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7493462 TI - Fluticasone propionate sensitivity in a patient with contact allergy to multiple corticosteroids. PMID- 7493463 TI - Allergic contact dermatitis from cyproheptadine hydrochloride. PMID- 7493464 TI - Occupational dermatitis from thebaine and codeine. PMID- 7493465 TI - Contact sensitization to topical salicylate. PMID- 7493466 TI - Spa pool dermatitis. PMID- 7493467 TI - Comparison of patch test results in 2 clinics reflecting different populations. PMID- 7493468 TI - Placebo-controlled oral challenge with cobalt in patients with positive patch tests to cobalt. PMID- 7493469 TI - Is tixocortol pivalate a photoallergen? PMID- 7493470 TI - Baboon syndrome due to mercury sensitivity. PMID- 7493471 TI - Allergic contact dermatitis from levobunolol eyedrops. PMID- 7493472 TI - Allergic contact dermatitis from plant extracts in a cosmetic. PMID- 7493473 TI - Human repeated insult patch test of 2-butoxyethanol. PMID- 7493474 TI - Acute irritant contact dermatitis from Agave americana L. PMID- 7493475 TI - Skin ulceration due to povidone-iodine. PMID- 7493476 TI - Occupational contact dermatitis from ceftiofur sodium. PMID- 7493477 TI - Tetrazepam: an allergen with several clinical expressions. PMID- 7493478 TI - Contact dermatitis and asthma from sodium metabisulfite in a photographic technician. PMID- 7493479 TI - Contact dermatitis from levobunolol and befunolol. PMID- 7493480 TI - Contact urticaria from litchi fruit (Litchi chinensis Sonn.). PMID- 7493481 TI - Occupational contact dermatitis from lichens and Frullania. PMID- 7493482 TI - Assessment of continuing sensitivity to MCI/MI (Kathon CG). PMID- 7493483 TI - Pigmented contact dermatitis. PMID- 7493484 TI - Allergic contact dermatitis from cyanoacrylate? PMID- 7493485 TI - Contact dermatitis due to a new fungicide used in the tulip bulb industry. AB - We describe an outbreak of contact dermatitis in a tulip bulb processing company. Shortly after the introduction of a new pesticide, the fungicide fluazinam, employees started to complain of dermatitis of the arms and the face. 8 employees were investigated and showed positive patch tests to fluazinam. The dermatitis disappeared quickly when they stopped work, but returned as soon they restarted. Subsequent investigations showed that the fungicide had not been used according to the manufacturer's recommendations. Fluazinam was shown to be a strong sensitizer under these circumstances. PMID- 7493486 TI - trans-inactivation of homologous sequences in Nicotiana tabacum. PMID- 7493487 TI - Gene silencing in transgenic plants: a heuristic autoregulation model. PMID- 7493488 TI - Paramutation in maize and related allelic interactions. PMID- 7493489 TI - DNA methylation and activity of the maize Spm transposable element. PMID- 7493490 TI - DNA methylation and transgene silencing in Petunia hybrida. PMID- 7493491 TI - Genetic and epigenetic inactivation of repetitive sequences in Neurospora crassa: RIP, DNA methylation, and quelling. PMID- 7493492 TI - MIP: an epigenetic gene silencing process in Ascobolus immersus. PMID- 7493493 TI - Gene silencing in Drosophila. PMID- 7493494 TI - Uptake of foreign DNA by mammalian cells via the gastrointestinal tract in mice: methylation of foreign DNA--a cellular defense mechanism. PMID- 7493495 TI - Transgene inactivation in Arabidopsis thaliana. PMID- 7493496 TI - Developmental regulation of co-suppression in Petunia hybrida. PMID- 7493497 TI - Suppression of flavonoid flower pigmentation genes in Petunia hybrida by the introduction of antisense and sense genes. PMID- 7493498 TI - Sense and antisense inactivation of fruit ripening genes in tomato. PMID- 7493499 TI - Co-suppression of beta-1,3-glucanase genes in Nicotiana tabacum. PMID- 7493500 TI - Proceedings of the 3rd annual scientific meeting of the Dysphagia Research Society, October 14-16, 1994, McLean, Virginia. PMID- 7493502 TI - Esophago/pharyngo/laryngeal interrelationships: airway protection mechanisms. AB - This presentation is part of a parallel seminar entitled "Esophago/Pharyngo/Laryngeal Interrelationships." It reports on some of the work done to elucidate the intricate mechanisms involved in transporting food/liquid through the pharynx while simultaneously protecting the airway from aspiration. Significant species differences occur during swallowing, which influence the interpretation of experimental results. Briefly discussed in this panel presentation are the distinction between the glottic closure reflex and laryngospasm; some variations that occur in normal and disordered adult swallowing; the sequence of events during swallowing; differences in the reciprocal relationship between breathing and swallowing in adults as contrasted with infants; and laryngeal reflexes engendered by esophageal stimulation. PMID- 7493501 TI - Airway protective mechanisms: current concepts. AB - There are at least eight mechanisms identified that result either in volume clearance of the pharynx and esophagus (secondary peristalsis and pharyngeal swallow) or prevent entry of the gastric content into the esophagus and pharynx (LES and UES), accentuate these barriers (esophago-UES and pharyngo-UES contractile reflexes), or induce closure of the vocal cords and introitus to the trachea (esophagoglottal and pharyngoglottal reflexes). The sum effect of various combinations of these mechanisms is suggested to help prevent retrograde aspiration. In other words, airway protective mechanisms against retrograde aspiration are multifactorial and involve delicate interaction between upper GI and upper airway tracts. Although the existence of these mechanisms in normal volunteers has been documented, their function in patients with retrograde aspiration and supraesophageal complications of gastroesophageal reflux disease currently awaits investigation. PMID- 7493503 TI - Pathological pharyngo-esophageal interactions. AB - Because of their normally coordinated function and physical contiguity, the pharynx and esophagus are subject to pathologic interactions in various disease or iatrogenic states. A useful conceptualization of these pathologic interactions is that of abnormal bolus transport. Specific clinical examples are given in which abnormal pharyngo-esophageal interactions appear to result in abnormal bolus transport, with its attendant consequences. PMID- 7493504 TI - Mechanisms of oral sensation. AB - Sensory nerves that supply mechanoreceptors in the mucosal lining of the oral cavity, pharynx, and larynx provide the substrate for a variety of sensations. They are essential for the perception of complex or composite sensory experiences including oral kinesthesia and oral stereognosis. Relevant to the concerns of the oral health care delivery specialist they also contribute to initiation of reflexes and coordination and timing of patterned motor behaviors. The response of oral mechanoreceptors to natural stimuli is determined to a large degree by morphological factors such as the nature of the relationship between nerve ending and certain cellular specializations, their distribution in the mucosa, the diameter of their primary afferent nerve fibers, and the central distribution of these fibers in the brainstem. Because of morphological similarities to certain cutaneous mechanoreceptors, the mucosal lining may be considered as an internal continuation of the large "receptor sheet" for localization and detection of mechanical stimuli. In some regions of the oral, pharyngeal, and laryngeal mucosa, this analogy is appropriate whereas in others, existing data suggest a different role consistent with regionally specific demands (i.e., initiation of protective reflexes). PMID- 7493506 TI - Cricopharyngeal myotomy may be effective treatment for selected patients with neurogenic oropharyngeal dysphagia. AB - The role of cricopharyngeal (CP) myotomy in the management of neurogenic oropharyngeal dysphagia remains controversial. A review of the literature regarding outcomes of CP myotomy for dysphagia in the setting of a variety of neurological disorders indicates a preponderance of favorable results. There are several potential explanations for reported improvement after CP myotomy for neurogenic dysphagia, including the possibility that it is an effective treatment, at least for selected patients. If this is true, appropriate selection criteria for this treatment of neurogenic dysphagia may include (1) intact voluntary initiation of swallowing, (2) adequate propulsive force generated by the tongue and pharyngeal constrictors, (3) videofluorographic demonstration of obstruction to bolus flow at the CP segment (rather than merely retention in the pharyngeal recesses), (4) manometric evidence of relatively elevated CP pressure in relation to the pharynx, and (5) relatively favorable neurological prognosis. The effectiveness and safety of CP myotomy for patients with neurogenic dysphagia are unlikely to be resolved without a prospective, controlled multicenter study enrolling patients who meet such criteria. PMID- 7493505 TI - Oropharyngeal dysphagia due to iatrogenic neurological dysfunction. AB - Oropharyngeal dysphagia due to iatrogenic neurological dysfunction may relate to either medication side effects or surgical complications. There are several general mechanisms by which neurological side effects of medications can cause or aggravate oropharyngeal dysphagia. These include decreased level of arousal, direct suppression of brainstem swallowing regulation, movement disorders (dyskinesias, dystonias, and parkinsonism), neuromuscular junction blockade, myopathy, oropharyngeal sensory impairment, and disturbance of salivation. Postsurgical oropharyngeal dysphagia due to neurological dysfunction has been described in association with carotid endarterectomy, esophageal cancer surgery, anterior cervical fusion, and ventral rhizotomy for spasmodic torticollis. A potential explanation for oropharyngeal dysphagia following these surgical procedures is intraoperative mechanical disruption of the innervation of the pharyngeal constrictor muscles by the pharyngeal plexus. Posterior fossa and skull base surgery can lead to dysphagia as a result of intraoperative damage to brainstem centers and/or cranial nerves involved in swallowing. Perioperative stroke is the most likely explanation for oropharyngeal dysphagia appearing acutely following surgery, especially if the type of surgery predisposes to embolism or hypoperfusion. PMID- 7493507 TI - Efficacy of behavioral treatment for oropharyngeal dysphagia. AB - In this critique of research in behavioral treatment of dysphagia, three types of investigations are briefly reviewed: general management guides, specific behavioral treatments applied to single patients or a few patients, and dysphagia programs applied to large numbers of patients. Efficacy of treatment has been demonstrated in some of the single-case or small-group studies where specific techniques have been utilized. Unfortunately, efficacy has not been demonstrated in any of the studies where large groups of patients have been enrolled in dysphagia programs. Some of the critical factors that have been lacking in many of these studies are discussed and the need to include functional outcome measures in efficacy studies is emphasized. PMID- 7493508 TI - Efficacy in dysphagia. AB - Data demonstrating the efficacy of behavioral methods for treating dysphagia are in short supply. This paper defines efficacy and distinguishes it from efficiency. It highlights the attitudes, measures, decisions, and data essential to the design of clinical trials. The need for additional outcome measures and for establishing appropriate treatment intensities are emphasized. PMID- 7493509 TI - Classification of eating impairments based on eating efficiency in children with cerebral palsy. AB - Eating impairments in children with cerebral palsy (CP) may vary widely from mild to severe. Accurate diagnosis of the severity of eating has been hampered by the lack of a classification system which would permit stratification of this wide range of problems into mild, moderate, and severe. We propose such a classification system based on measures of (1) growth: weight, height, and skinfold thickness and (2) eating skills: eating efficiency and oral-motor skills. One hundred children, 54 boys and 46 girls from 2 to 16 years, who had various degrees of CP and eating impairments, were studied. Classification, treatment effectiveness, as well as the implications for growth monitoring, based on this classification system are discussed. system are discussed. PMID- 7493512 TI - [Arterial vascularization of the sinoatrial and atrioventricular node (preliminary study apropos of 15 anatomic parts]. AB - Arterial supply to the sino-atrial and atrioventricular node is studied after coronary RHODOPAS injection on 15 adult hearts taken off during autopsy. Both coronaries were injected in 7 cases, left ostium in 2 cases and the right on in 6 cases. The sino-atrial node is supplied by the right sino atrionodal artery (S.A.N.A) in 6 cases upon 7 and in 3 cases by the left S.A.N.A. Anastomoses exist in 2 cases between the right S.A.N.A. and the left S.A.N.A. The atrio-ventricular nodal artery arises from the retro-ventricular trunk branching from the right coronary artery mainly. The authors emphasize that care must be taken to protect the vascular supply to the conduction tissue during cardiac surgery. PMID- 7493511 TI - Dysphagia after pharyngolaryngeal cancer surgery. Part II: Implications for reconstructive procedures. AB - In the base of the tongue region, reconstructive procedures have to provide more bulky-tissue coverage (i.e., myocutaneous flaps) in order to avoid cranial release of pressure and to bring about swallowing initiation. Resections of the pharyngoesophageal (PE) segment cause circular defects, always affecting the sphincter and necessarily relaxation, thus reducing the hypopharyngeal suction pump. The resistance to bolus flow, therefore, is generally increased but can be compensated by a stronger tongue driving force. In addition to the functional obstruction, special attention is called to the growing lumen discontinuity between the wide pharynx and the narrow esophagus. Plastic reconstructions, therefore, have to compensate for different lumina distally. Following ablative surgery in the upper esophageal sphincter region, a softer and smoother tissue coverage is warranted in order to facilitate bolus transfer to a passive bolus flow if necessary. For that purpose, a new myofascial pectoralis flap was designed based on morphometric investigations and postmortal selective injection studies. In this flap, the bulky muscle mass is separated from just a vascularized, thin fascia-muscle layer. The donor site is covered with the remaining bulky muscle-skin complex left intact. The fascial flap covers defects where a soft lining is required and replaces the PE segment as a tubed neopharynx. Histologic specimens show a reepithelization with local mucous membrane from the anastomotic site to the fascial surface. The resistance to bolus flow is reduced, thus alleviating the tongue driving force, which is increased for compensation in any case. PMID- 7493510 TI - Dysphagia after pharyngolaryngeal cancer surgery. Part I: Pathophysiology of postsurgical deglutition. AB - Eighty-one patients were examined after laryngopharyngeal cancer surgery with a sequential computer manometry system using 4-channel-pressure probes. The general swallowing coordination is neither a matter of the oropharyngeal pressure thrust nor of the pharyngeal transit time, but mainly depends on swallowing initiation. The points of interest are both the pharyngeal inlet and outlet. The topographic correlates are the base of the tongue and the upper esophageal sphincter (UES). Resections of the base of the tongue lead to a decrease of volume available for pressure generation, thus reducing the tongue driving force. The swallowing reflex is uncoordinated resulting in dyskinesia of the UES. Compensation may be achieved with a stronger oropharyngeal thrust and/or repeated swallows. Distal resections alter the pharyngoesophageal segment so that a functional obstruction results, combined with lower pressure amplitudes in the hypopharynx, reducing the pressure gradient necessary for bolus flow. This increasing resistance can be overcome by higher propulsive forces in the base of the tongue region. In case of additional lingual defects, deglutition is subject to decompensation, highlighting the major role of the tongue as a pressure generator for bolus passage. PMID- 7493513 TI - [Epidemiologic study of the causes of esthetic restorations in pedodontic clinics in Dakar]. AB - The authors expose a retrospective study carried out on records of patients from 18 months to 15 years old consulting for esthetic restorations in 3 dental health care centers of Dakar. The total number sampled is 151 children with 89 boys and 62 girls representing 3.5% of the total number of consultants [3964] during two years [1991 and 1992] in the dental health care centers. The study shows that the main etiological factors of the esthetic restorations in constant progression, are dental traumatisms, and caries infections. It shows also that the request varies with age and sex and that the male are more represented in the sample. Face to the lack of structures, material and human means, information, education and communication for the oral and dental health have to be fully broadcasted in the kindergartens and elementary schools in the limit of an entire prevention program in order to reduce the needs in this precise field. PMID- 7493514 TI - [Genetic and etiologic aspects od spina bifida in Senegal. Apropos of 211 cases collected at the neurosurgical clinic UHC of Fann]. AB - From 1969 to 1988, 211 cases of spina bifida were collected from the Neurosurgical Clinic at the University Teaching Hospital of Dakar. Cases were reviewed for genetic and environmental factors. There were 103 males (48.81%) and 108 females (51.19%). The most important ethnic group involved were toucouleurs (20.11%). Consanguineous marriages in parents of affected infants were lower than in the general population (62.5% vs 71%). Recurrence has been noted in 4.26% of the cases. There were more fifth-born children affected than expected (16.2% vs 9% p < 0.02). A seasonal and geographic variation of the incidence has also been observed. Infants conceived between November and February appear to be of highest risk (36.16%). The incidence is higher in the north-west and lower in the south east part of the country. Lower social-economic classes were the most involved. The authors support the hypothesis of multifactorial etiology in Spina Bifida. PMID- 7493515 TI - [Attitude and behavior of the youth of Senegal toward drugs]. AB - Formerly known as a dreadful calamity for industrialized countries, drug abuse now seems to have spread all over the world. In Senegal, it has been very difficult to gather reliable statistics on its extensiveness. To better define the problem in Senegal, UNESCO has launched an epidemiological study on young people drug abuse and alcoholism in schools and elsewhere. The authors analyse the results of this study and draw the following conclusions: all types of drug use are linked to religious practice, drug use by young people is due to curiosity or their ignorance of drug drawbacks and their lack of education, school, family, religious education and social repression play an important role in the fight against drug use. PMID- 7493516 TI - [Anatomy of coronary arteries]. AB - To study the coronary arteries we collected 200 black african hearts which coronaries have been injected with Rhodopast then corroded. Most of the arteries (74.10%) originate from two separate sinuses one on the left and the other one on the right. The right coronary artery runs in two different modes. In one mode it is short and ends in different branches. In the other mode it is long and runs up to the crux. The left coronary artery arrives on the left border of the heart and gives the circumflex and the left border artery. The bifurcation mode of division was 32.25% for the right coronary artery and 69.35% for the left coronary artery which had a trifurcation in 36.66%. The superior atrial artery was not seen in 3 cases and the infundibular arteries derived from the aorta in 3 cases. We have encountered intercoronary anastomoses in 82.6% of cases. These results are similar to these observed in the literature. The only difference is on the rich intercoronary anastomosis which seems to be an african particularity even though we cannot establish if they are functional or not. PMID- 7493517 TI - [Anatomo-clinical aspects of ganglionic tuberculosis in Senegal: comparative study between the 60's and 80-90's apropos of 139 cases]. AB - The authors report the results of a retrospective and comparative study of anatomo-clinical aspects of ganglionic tuberculosis carried out over two periods of five years each: first period: 1962-1966; second period: 1988-1992. This study shows a higher prevalence of this disease among children during the first period with an odd ratio of 4.36. In the second period, young adults are the most affected particularly the age group between 30 and 39. The possible role of the HIV virus in this phenomenon has been pointed out. The most common histological forms were the subacute ones with a distinct prevalence during the second period, on the opposite of the acute and chronic forms. PMID- 7493518 TI - [Echographic and velocimetric aspects of arteriopathies in the diabetic]. AB - AIMS: Study of Diabetic arteriopathies Arteriopathies in cervico encephalic arteries and lower limbs were detected both with Echography and Doppler flow velocity waveform analysis from 60 treated patients. They were shared into 34 women (57%) and 26 men (43%). Patients age varied between 39 and 77 years. 85% were non insulin dependent versus 15% insulin dependent patients. 36 patients (60%) presented arterial abnormalities, 14 of which (23.33%) were located in the lower limbs, 19 (31.66%) in the carotid arteries, and 3 (7%) in both carotid arteries and the lower limbs. Arteriopathies were found in medium aged patients (40-60 years). Arteriopathies are often noted in medium aged diabetics. The prevalence of lesions were relatively higher in supra-aortic trunks (36%) than in lower limbs arteries (28%). They are distal in lower limbs and constitute a factor of diabetes prognosis. The sensitivity and the less expensive cost of ultrasonor methods give a first choice to these technics in the patients following up and prevention of vascular complications of diabetes in Africa. PMID- 7493519 TI - [Essential atrophy of the iris apropos of 1 case]. AB - The authors report a case of bilateral essential iris atrophy syndrome in a twenty two years old man associated with unilateral cataract and glaucoma. They stress the rarity of the syndrome (first case in their section) and the uncommon bilateral case. The cataract unusually described in the syndrome would be directly linked to the important and durable ocular hypertension. PMID- 7493521 TI - [Amoebic liver abscess: echographic aspects]. AB - Amoebic liver abscess is the most frequent location of the extra-intestine amibiasis with an epidemio-endemic repartition in our areas. We are reporting in this study the main echographic patterns that can be found. 117 documents were collected and studied between 1982 and 1988 in the main hospitals of Dakar (SENEGAL). Most of the patients were young, the range of age being between 25 and 55 years old and 83% of them, were male. The diagnosis of the amoebic liver abscess was evocated on the basis of the following clinical and biological symptoms: 54.38% of painful haetomegaly, 42.10% of pleuro-pulmonary and digestive signs, 3.50% of long lasting isolated fever, non specific biological sign of inflammation, 74.57% of positive hemaglutination test. An echographic test was performed before the anti-parasitic treatment with an echotomograph PHILIPS SDR 1500 in real time using a probe of 3 MHZ. The amoebic abscess of liver was detected by the echography in all cases. The unique abscess (83.10%) was the most frequent form. It was localized in the right liver (64%) and had an heterogeneous echostructure (55.70%). The hypo-echogeneous form (36.50%) was the earlier stage of the collecting abscess. The liquid form (07.80%) was observed in the latter stages of the disease. Some difficulties to determine the amoebic abscess may appear when primitive liver cancer or pyogensus abscess are present. In these cases it is necessary to analyse the liquid of ponction to be affirmative. PMID- 7493520 TI - [HIV infection: a new concern in dental public health]. AB - The authors report the results of a seroepidemiological study, conducted in Senegal on dental personnel. This study allows them to confirm that professional dental practice is a factor of risk for the HIV infection. This study recommends work methods to the dental profession. PMID- 7493522 TI - [Non typhoidic salmonellosis in the African pediatric population]. AB - The study of non-typhoidic salmonellosis in A. royer pediatric Hospital (Dakar) during a five year term (1985-1989) was realised on thirty five medical records. This pathology accounts for 0.4% of admissions and occurs mainly on children under 3 years old (83%) essentially with a pathologic background. The main clinical forms were septicemia (46%) and gastro-enteritis (31%). Purulent meningitidis represented 45% of all the localized forms. Only one case of asymptomatic carriage has been identified. The seventeen serotypes of Salmonella identified belong to eight serogroups. S. enteritidis and S. typhimurium represented 51% of the isolates. Susceptibility to antibiotics of these different serotypes were variable, the third generation cephalosporins and gentamycin having inhibited more than 80% of the strains. The mean duration of antibiotherapy was 21 days with a mortality rising to 17%. The association ampicilline-gentamycin although criticable, remains indication as first treatment because of the availability of these drugs. PMID- 7493523 TI - [Intestinal parasitic diseases and tinea of the scalp in Dakar school population: influence of environmental factors on the infestation level]. AB - We carried out a study during February 1993 in a french and three coranic schools in order to evaluate the incidence of the economical level on the prevalence of intestinal parasites and tinea's scalp. Infection rate by intestinal parasites were 67.4% and 84% respectively in french and coranic schools with significant differences: X2 = 10.8. Infection rate by dermatophytes were 10.6% and 48.8% respectively in francophone and coranic schools with significant differences: X2 = 29.3. Ascaris was predominant through all ages in coranic schools, while amaeba were the main parasites in the upper ages in the french school. Bad environmental features observed in coranic schools as well as in popular areas are risk factors to the infection and contribute to increase the prevalence of intestinal parasitic diseases and tinea's scalp. PMID- 7493524 TI - [Place on information in health services use by mothers in Guediawaye]. AB - The authors report the results of a survey in a suburban healthcare structure. The purpose was to evaluate the knowledge of mothers about the healthcare delivered through a childhood and maternal healthcare program in Senegal. 244 women were selected at random. The consultant mothers came more for their children than for themselves (79% of consultations were for their children, 49% for themselves). 60% to 80% of women are unaware of basic notions currently teached through the "Information-Education-Communication (I.E.C)" program. All that requires to elaborate with these women healthcare strategies, which are in adequacy with their needs in these peripheral healthcare structures, and which are supported by an adequate and regularly evaluated I.E.C. PMID- 7493525 TI - [Ovarian pregnancy apropos of 2 cases]. AB - The authors report 2 cases of ovarian pregnancy at Gynecological Obstetric Clinic of University Teaching Hospital in Dakar out of 210 ectopic pregnancy registered in a 28 month period (March 91 to June 93). These are the fourth and fifth cases operated on in Dakar (the 3 first cases are reported during the last 10 years). In light of the 2 cases using the data of the literature, the authors try to find out the particularities of the scarce affection which etiopathogeny is still unclear even if the symptomatology is often similar to a tubal pregnancy. The diagnosis is often evoked during laparatomies but its confirmation is based exclusively on the anatomopathological test. The frequent existence of intra ovarian hematoma and zones of necrosis make people practise annexectomy which should be avoid as much as possible. PMID- 7493527 TI - [CHolelithiasis in Burkina Faso (apropos of 44 cases operated on at NHC YO]. AB - The authors relate 44 cases of cholelithiasis, operated on between 1986 and 1990 at the General and digestive Surgery Unit of the National Hospital Center YALGADO OUEDRAOGO of Ouagadougou. They established a progressive increase in the frequency of this affection thanks to the echographic diagnosis. The patients average age is 35 years old and paradoxically, the sex-ratio is for men. It is about lithiasis with some predominant pigment constituent, located in the vesicule area in 98% of the cases, associated in 18% with a localization in the common bile duct. The clinical symptomology is atypic. But the complicated forms with acute cholelithiasis, or with a big cholostasis liver and dilatation of the common bile duct, are not rare. The preoperative cholangiography is not used. All this resulting in frequent surgical procedures on the common bile duct. Anyway, the therapeutic results are satisfactory: the surgery death rate is 2.27% and there are few complications. PMID- 7493526 TI - [Phospholipid levels in exploration of cerebral atherosclerosis in African Senegalese population]. AB - In the investigation of atherosclerosis, cholesterol and its fractions, triglycerides and glucose are more tested than phospholipids. We were interested by this category of lipids in Senegalese suffering from cerebrovascular disease. We did not find differences between healthy and non healthy subjects for the total phospholipids; but the high density lipoprotein phospholipids were significantly lowered in non healthy subjects. This decrease might explain in part, the atherosclerosis strike. Practicians should pay more attention in this lipidic parameter, using the reference values established in Senegalese. PMID- 7493528 TI - [Congenital glaucoma (apropos of 58 cases)]. AB - In a retrospective study on 10 years, the authors analyse epidemiological clinical and therapeutic aspects of congenital glaucoma. From 1979 to 1989, 58 cases of congenital glaucoma were found in children between 0 and 12 years, of which 53 bilateral and 5 unilateral cases (111 eyes). The frequency of 0.10% is considered high. According to studies carried out by Europeans, it is estimated between 0.01 and 0.024%. The authors stressed the important rate of consanguinity (46.5%) and masculine prevalence (74.1%). Disappointing therapeutic results and bad prognostic depend on the advanced cases of clinical forms found during the first consultation. The authors advocate preventive measures by genetic counsel and an early surgical treatment for best functional results. PMID- 7493529 TI - [Growth hormone, a potential way to improve livestock productivity in Africa]. AB - This paper reviews experimental data on exogenous pST administration effects on growth and carcass performances in pigs to show the importance pST could have for pigs breeding in Africa. In Europe and USA pST breeding conditions induced growth rate increase (+18.7%) and a decrease of the feed intake (-14.6%) and the feed/gain ratio (-26.5%). The carcass muscle percentage was increased (+17.8%) and the fat rib was decreased (-25.6%) in treated animals compared to controls. pST induced a generalised thoracic and abdominal organs hypertrophy which can partly explain the slight decrease of dressing percentage. On the meat quality pST didn't have any pronounced detrimental effect. According to these interesting results, pST effects must be tested in Africa breeding conditions. PMID- 7493530 TI - [Caudal epidural anesthesia in 70 children]. AB - Caudal loco regional anesthesia is a quite simple and secure procedure. That is a prospective study made on seventy children operated in orthopaedics, urology and general surgery below the ombilicus. The caudal epidural anesthesia followed a general anesthesia in all seventy cases and brought relief in terms of analgesia, even for those painful surgical procedures. The good post operative status with only some minor complications leads us to use widespreadly caudal epidural. PMID- 7493531 TI - [Toxicity of cholestatic bile acids on intrahepatic biliary cells of the rat]. AB - The aim of this study is: 1. to isolate intrahepatic biliary epithelial cells and, 2. to study the toxicity of cholestatic biliary acids on these cells. Cells were isolated from rats with proliferated bile duct-cells, that were induced by a 21 days bile duct ligation. They were characterized by glutamyltranspeptidase and cytokeratins 7 and 19 using histochemical or immuno cytochemical methods. Isolated cells were incubated with different concentrations (250, 500, 1000 and 2000 microM) of cholestatic biliary acids, lithocholate sulfate (LCS) and taurolithocholate sulfate (TLCS. Cells viability is estimated by three methods: Trypan blue, LDH release and MTT test. We obtained purified fractions of non parenchymal liver cells enriched in biliary epithelial cells (> 95%). On these cells, we showed toxicity of LCS and TLCS and determined CI 10 and CI 50 of these acids which were respectively 800 microM and 2 mM for LCS; 1.4 and 2 mM for TLCS. These results indicate that cholestatic biliary acids (LCS and TLCS) are toxic for biliary cells. This cytotoxicity can be probably a possible mechanism of cholestasis. PMID- 7493532 TI - Long-term results of low vision rehabilitation in age-related macular degeneration. AB - One to five years after successful low vision rehabilitation, 199 patients with age-related macular degeneration were asked by means of a questionnaire whether they still used their low vision aids and whether they were satisfied with the low vision counseling. We received feedback from 66% of the patients. 20 patients had already passed away. 74% of the 112 answering patients still use a low vision aid today. Three-quarters of them still use the originally prescribed low vision aid. Interestingly enough, the daily reading time was rather short, only 16% of the patients read more than 1 hour per day, 42% read somewhat more and 13% somewhat less than 10 minutes per day. Retrospectively, 81% of all responding patients found their low vision counseling at least a little, more of them quite or very helpful. These numbers suggest, that the initial labor of patiently examining and training older, visually handicapped patients is worthwhile. PMID- 7493534 TI - The optics of the eye-lens and lenticular senescence. A review. AB - Although the lens of the eye is structurally a biological tissue, it functions as an optical element providing one third of the refracting power of the human eye, and a variable focus in younger years. Throughout a life-time the optical properties of the eye-lens alter, resulting in changes in function: there is a gradual depletion of the focussing amplitude from infancy to middle age, and a loss of transmittance in the later decades of life. The optical properties of the lens depend on its power, which in turn is determined by its physical dimensions (curvatures and thickness) and its refractive index as well as transmissivity and the organization of its internal components. The power of the functional lens is, however, modifiable by virtue of the lens being attached via the zonule to the ciliary muscle. The contraction and relaxation of the latter respectively increases and decreases lens power in accordance with innervations determined by the physical distance of external objects to be imaged on the retina. This review will consider many of these features and how alterations in any of them may lead to changes in lenticular function. However, as we have recently devoted a detailed study to presbyopia [1] its mechanism will not be considered here. PMID- 7493533 TI - Retinal arteriovenous communication in retinitis pigmentosa with Refsum's disease like findings. AB - In this report we describe, herewith, a patient with primary pigmentary dystrophy of the retina (retinitis pigmentosa) associated with unilateral retinal arteriovenous communication and exudative retinal detachment. The patient had complete resolution of the retinal detachment following laser photocoagulation treatment. Such association has not been previously reported. PMID- 7493535 TI - Vascular tumors of the orbit. AB - Eighty-five vascular lesions of the orbit examined and treated between 1963-1993 were reviewed retrospectively to reveal the types of vascular tumors, age and sex distribution, clinical characteristics, treatment options and prognosis. Capillary hemangioma was the most frequent orbital vascular tumor accounting for 37 of 85 cases making up 43.5% of the entire orbital masses. Cavernous hemangioma accounted for 35 cases (41.2%), varices for 3 cases (3.5%), anteriovenous malformations for 3 cases (3.5%), angiosarcomas for 3 cases (3.5%), lymphangiomas for 3 cases (3.5%) and hemangiopericytoma for one case (1.2%). Of the 85 vascular tumor cases 43 were encountered in children aged 18 years or less. Capillary hemangioma was the most frequent pediatric orbital vascular lesion, making up 86% of the vascular tumors in this age group. Capillary hemangiomas have been treated with observation and intralesional corticosteroid injections. Cavernous hemangiomas, vascular malformations, lymphangiomas and two cases of angiosarcomas received orbitotomy. One case each of angiosarcoma and hemangiopericytoma underwent exenteration because of the advanced stage of the lesion. There was one tumor-related death who was a 9-year-old boy diagnosed with angiosarcoma. Vascular tumors with the exception of angiosarcomas and hemangiopericytoma behaved as benign lesions. PMID- 7493536 TI - Granulomatous anterior uveitis presenting with acute posterior multifocal placoid pigment epitheliopathy. AB - Acute posterior multifocal placoid pigment epitheliopathy (APMPPE) has been described as a posterior segment inflammatory disorder of young adults of unknown etiology. Granulomatous anterior uveitis in association with APMPPE has only rarely been reported in the literature. We report a patient who presented with clinical and angiographic findings consistent with APMPPE in addition to granulomatous anterior uveitis with mutton-fat keratic precipitates and Koeppe nodules. This latter finding has not been emphasized as a possible feature of APMPPE. PMID- 7493537 TI - Evaluation of the range of areas of the fluorescein staining patterns of the tarsal conjunctiva in man. AB - Fluorescein dye is commonly used to highlight the tarsal conjunctiva in man to facilitate diagnosis of papillary conjunctivitis. A quantitative analysis of the fluorescein-highlighted features could be useful in both assignment of severity grades as well as for objective comparisons but no data is available on the distributions of sizes (areas) of these features. An exploratory, range-finding study was undertaken to provide data on feature areas that might be encountered. After application of fluorescein, 35 mm photographs were taken at a fixed distance with zone 1/zone 2 (Allansmith) of the tarsal plate aligned perpendicular to the camera. The photographs were projected, an overlay of 150 to 180 contiguous features outlined by fluorescein made and planimetry used to assess the areas and area distributions of the features on tarsal plates clinically graded with stages 1 to 4 papillary hypertrophy. For zone 1 of the tarsal plate in asymptomatic subjects, a fluorescein solution highlights the limits of features that enclose an area averaging 30,000 microns2 (range 8000 to 120,000; median area of close to 35,000 microns2). The areas of these features were generally normally distributed and there was little variation in feature areas across the zone. At different stages of clinical papillary conjunctivitis, the areas of the negative staining patterns are larger. Averages of 60,000 to 90,000 microns2, median values of close to 70,000 microns2 and individual feature areas up to 350,000 microns2 were seen in mildly symptomatic patients. These distributions were generally heterogeneous and showed a skewed distribution. Areas averaging up to 0.755 mm2 (range 0.6 to 3.2 mm2) were measured in moderate to severe papillary hypertrophy. PMID- 7493538 TI - Asymmetry of motion VEP in infantile strabismus and in central vestibular nystagmus. AB - Norcia et al. [1] found a nasal-temporal asymmetry of visually evoked potentials (VEP) elicited by motion stimuli in patients with infantile strabismus. Patients with infantile strabismus typically present with an asymmetry of the monocular optokinetic nystagmus (OKN). We here address the question whether the asymmetry of the motion VEP indicates a sensory defect in the afferent visual pathway that could explain the OKN asymmetry. We recorded the VEP to a horizontally oscillating vertical sinusoidal grating in 20 patients with infantile strabismus (esotropia, asymmetry of the monocular optokinetic nystagmus, latent nystagmus) and in 10 normal controls. No asymmetry occurred in the 10 controls. Eight of the 20 patients with infantile strabismus showed a clear difference between the VEPs evoked by back and forth movements with a mirror-like asymmetry between the two eyes (phase shift 180 +/- 20 degrees). However, there was no significant correlation between the degree of VEP and OKN asymmetries. Therefore, we assume that the VEP asymmetry does not reflect the primary cause of the OKN asymmetry. Rather, the OKN asymmetry may be due to a sensory-motor defect in the efferent subcortical pathway, and the VEP asymmetry could be an epiphenomenon. Some of the VEP asymmetry may be a consequence of the latent nystagmus typically released under monocular stimulation, leading to adaptation of the afferent retino cortical pathway. This suggestion is supported by a marked VEP asymmetry that we found in two patients with an acquired central vestibular nystagmus, an abnormality most likely not combined with a primary defect of the retino-cortical pathway. PMID- 7493539 TI - HPR perimetry and Humphrey perimetry in glaucomatous children. AB - Fifteen children aged less than 11 years, affected by congenital glaucoma, underwent visual field testing using two different methods: conventional computerized perimetry (24-2 Humphrey program) and High Pass Resolution perimetry (Ring test). The aim of the study was to discover which of the two perimetric techniques was more suitable for younger patient. Results showed that HPR perimetry is more suitable to children. Reasons include the short duration of the test (5 min), and the appearance like a pleasant game to 80% of the subjects tested. In addition it proved to be better than Humphrey test because of the minimal possibilities of memorisation with a reproducibility factor of 68% in our sample and because of the reliable results and satisfactory evaluation in 76% of the eyes examined. PMID- 7493540 TI - Genetic features of retinitis pigmentosa in Turkey. AB - Sixty-two cases with retinitis pigmentosa from 42 index families were investigated to reveal the genetic features of the disease in Turkey. There were 42 propositi of whom 5 had a systemic syndrome associated with retinitis pigmentosa. Of the remaining 37 cases the condition was autosomal recessive in 21 (56.8%), sporadic in 12 (32.4%), autosomal dominant in 3 (8.1%) and X-linked recessive in one (2.7%). Sporadic cases may be more frequent as many hereditary cases are not brought to medical attention in rural families. Male preponderance among sporadic cases may indicate that there may be more X-linked cases. Nine out of 21 cases initially classified as sporadic displayed parental consanguinity and they were included as having autosomal recessive trait. Large families with autosomal recessive inheritance may prove valuable in linkage analysis and in defining future gene abnormalities. PMID- 7493541 TI - Glaucoma triple procedure comprising trabeculotomy with a fornix-based scleral flap. AB - A new triple surgery procedure was applied to 20 primary open-angle glaucomatous (POAG) eyes. This technique comprises trabeculotomy with narrow (2-3 mm) fornix based scleral flap, combined with extracapsular cataract extraction (ECCE) and posterior chamber intraocular lens (PC-IOL) implantation. As to surgical results, at two years postsurgery 53% (9/17) of the eyes were controlled under 21 mm Hg without medication, 35% (6/17) with topical medication alone. Eighteen (90%) eyes showed improved visual acuity of two or more Snellen lines. No serious postoperative complications occurred. There was no significant difference in postoperative astigmatism as compared with ordinary ECCE and PC-IOL surgery. Trabeculotomy with fornix-based scleral flap offers the advantage of simple and safe surgery when combined with ECCE and PC-IOL implantation. PMID- 7493542 TI - Effect of liver intoxication by carbon tetrachloride on hepatic local disposition of oxacillin using moment characteristics as index. AB - The effect of liver intoxication by carbon tetrachloride (CCl4) on the hepatic local disposition using oxacillin as a test drug and bovine serum albumin (BSA) as a reference substance was evaluated by single-pass bolus input method in the isolated perfused rat liver. Oxacillin and BSA were introduced into the liver from the portal vein, and the outflow concentration-time profiles of oxacillin and BSA from the rat liver into the hepatic vein were kinetically assessed by moment analysis. Liver damage was monitored by plasma SGOT, plasma SGPT, and bile flow rate. Hepatic recovery ratio FH of oxacillin increased from 50% to 80%, with an increase in liver intoxication by CCl4; whereas FH of BSA was naturally 100%. Mean transit time tH of oxacillin increased from 6 to 12 sec, with an increase in liver intoxication; whereas tH of BSA increased from 7 to 10 sec. The relative variance sigma 2/tH2 of oxacillin increased from 0.2 to 0.7, with an increase in liver intoxication; whereas sigma 2/tH2 of BSA took the value of approximately 0.4, independent of liver damage. In the relationship between the dispersion model and moments, it was shown that the blood space VB increased from 15 to 20%, the index for distribution (1 + k') of oxacillin from 1.0 to 1.5, the efficiency number RN of oxacillin decreased from 0.7 to 0.3 with the increase in liver damage, and the extent of eddy mixing was predicted to be unaffected by liver damage. The intensive increases in tH and sigma 2/tH2 of oxacillin with the liver intoxication by CCl4 were explained by increases in the extent of distribution and nonequilibrium distribution, respectively.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7493543 TI - Mechanism-based inactivation of rat liver cytochrome P4502B1 by phencyclidine and its oxidative product, the iminium ion. AB - Cytochrome P4502B1, the major phenobarbital-inducible isozyme in the rat liver, is inactivated by phencyclidine (PCP). Incubation of PCP with purified P4502B1 in the reconstituted enzyme system with NADPH-cytochrome P450 reductase and phospholipid resulted in a marked loss of activity as measured using a secondary incubation mixture for 7-ethoxycoumarin O-deethylase activity. The loss of activity required NADPH and PCP, and the activity decreased in a time-dependent, pseudo-first-order process indicative of mechanism-based inactivation. The rate constants for inactivation were dependent on the PCP concentrations and displayed saturation kinetics. A KI = 3.8 microM and kinact = 0.12 min-1 were determined for the inactivation by PCP. The partition ratio calculated from a plot of the percentage activity remaining after 45 min vs. the concentration ratios of PCP to P450 was 45. Although 90% of the catalytic activity was lost after a 45-min incubation, little loss was seen in the optical spectrum at 418 nm or in the ability of the reduced enzyme to bind CO. The inactivation was not inhibited by the addition of cyanide, whereas substrates such as 7-ethoxycoumarin protected against the inactivation. The iminium ion of PCP, an oxidative metabolite, inactivated P4502B1 in the same fashion as PCP. These results demonstrate that PCP is an efficient mechanism-based inactivator of rat liver P4502B1 and does not inactivate by modification of the heme moiety. PMID- 7493544 TI - Interaction between propranolol and amino acids in the single-pass isolated, perfused rat liver. AB - Propranolol (PL) bioavailability has been shown to increase substantially when it is administered with a protein-rich meal. A change in metabolic capacity or tissue uptake, induced by amino acids (AAs) released as a result of digestion of dietary protein, is a possible contributing mechanism to the food effect. This hypothesis was tested in isolated, perfused rat livers in the single-pass mode. Rac-PL (20 micrograms/ml) was infused to steady-state at 3 ml/min/g liver for 150 min. A balanced mixture of I-AA was coinfused from 70 to 110 min. The AA reversibly increased the steady-state concentration of PL by 18% and reduced steady-state concentrations of 4-hydroxypropranolol, N-deisopropylpranolol, PL glycol, naphthoxylactic acid, and naphthoxyacetic acid by an average of 41% and propanolol conjugates by almost 100%, indicating metabolic inhibition. In a second experiment, PL was coinfused with AAs from the beginning of the experiment, and tissue binding was compared with control livers. There was no significant effect of AAs on PL tissue binding. In a third study, the effect of four different concentrations of AAs coinfused from 70 to 110 min was assessed. The percentage change in PL and phase I metabolite levels was linearly correlated to the influent AA concentration. The large magnitude, reversibility, lack of pathway specificity, and concentration dependence of the AA interaction in the perfused liver are also features of food interaction in humans. These similarities constitute evidence that metabolic inhibition by AAs originating from dietary protein could contribute to the PL-food interaction. PMID- 7493545 TI - Characterization of the metabolites of the peptidomimetic human immunodeficiency virus type 1 protease inhibitor SK&F 107461 in rats using liquid chromatography/mass spectrometry. AB - The metabolic fate of SK&F 107461 [Cbz-Ala-Ala-Phe psi [CHOHCH2] Gly-Val-Val OMe], a potent and specific inhibitor of the protease encoded by human immunodeficiency virus type 1, in male Sprague-Dawley rats is described. SK&F 107461 is a hexapeptide analog containing a hydroxyethylene linkage in place of one of the peptide bonds, and in which the amino terminus is blocked with a carbobenzyloxy group and the carboxy terminus is modified to a methyl ester. The major metabolites of SK&F 107461 found in bile and urine after intravenous administration of 3H-labeled compound were characterized by LC/MS using either thermospray or continuous flow/FAB models of ionization. Approximately 80% of the administered radioactivity was recovered in the bile of bile duct-exteriorized rats following an intravenous dose. Radiochromatographic profiling indicated that SK&F 107461 was subject to extensive biotransformation. Structures were determined for three major biliary and five major urinary metabolites. Two of the major circulating plasma metabolites observed after intravenous bolus administration had similar retention times to metabolites that were observed in both bile and urine. A pathway for the biotransformation of SK&F 107461 in the rat is proposed. The parent molecule underwent two primary modes of metabolism. Hydrolysis of the carboxy-terminal ester or hydrolysis of the Ala-Ala peptide bond near the amino terminus were the primary metabolic events. All of the other metabolites characterized can be accounted for by exopeptidase activity subsequent to one or both of these primary events. There were no major metabolites observed resulting from anything other than hydrolysis of the ester or peptide bonds in the parent molecule. PMID- 7493546 TI - Human metabolism of dolasetron mesylate, a 5-HT3 receptor antagonist. AB - Dolasetron mesylate is a selective and potent 5-HT3 receptor antagonist. This drug is currently in development for the treatment of nausea and vomiting in chemotherapy. The metabolism of dolasetron mesylate was studied in six healthy male volunteers who were given a single 300 mg oral dose of [14C]dolasetron mesylate. An average of 59% of the total radioactivity was recovered in the urine and 25% in the feces. Metabolites were quantitated in urine samples taken up to 36 hr postdose. Reduced dolasetron (RD) accounted for 17-54% of the dose in urine. Hydroxylated metabolites of RD made up no more than 9% of the dose in urine. Most of the remaining urinary radioactivity consisted of conjugated metabolites of RD and hydroxy RD. Hydrolysis of selected urine samples showed that the glucuronide of RD was the most abundant conjugate in urine. A small percentage of the dose (< 1%) in urine was identified as the N-oxide of RD. Analysis of urine samples by chiral HPLC indicated that the R(+):S(-) ratio of RD was approximately 9:1. PMID- 7493547 TI - Structural determination of a glucuronide conjugate of flucytosine in humans. AB - The structure of a glucuronide metabolite of flucytosine (FC; 5-fluorocytosine), found in the urine of all patients treated with this antifungal drug, was determined. This compound is the O2-beta-glucuronide of FC. Its structure was established after isolation from urine and by comparing its spectroscopic characteristics with those of three FC glucuronides previously synthesized. This study is the first report of the identification of a glucuronide of a fluoropyrimidine drug in humans. PMID- 7493548 TI - 3-Methylcholanthrene and pyridine effects on CYP1A1 and CYP1A2 expression in rat renal tissue. AB - The effects of 3-methylcholanthrene (3-MC) and pyridine on rat renal cytochrome P450 (CYP) 1A1 and 1A2 mRNA expression have been examined by Northern-blot analysis and reverse transcriptase-polymerase chain reaction (RT-PCR), followed by Southern-blot analysis of the PCR products. Northern-blot hybridization and RT PCR analyses of kidney poly(A)+ RNA revealed that 3-MC treatment produced a time dependent increase in the renal CYP1A1 and 1A2 mRNA levels, with CYP1A1 and 1A2 mRNA levels maximally increased at 24 and 18 hr, respectively, after treatment. These data were confirmed via RT-PCR analysis using a subsaturating level of cDNA template and by Southern-blot analysis of the PCR products. This approach served as the foundation for examining the effects of pyridine on CYP1A1 and 1A2 expression in renal tissue. RT-PCR analysis of renal CYP1A1 and 1A2 poly(A)+ RNA levels after treatment with pyridine (200 mg/kg/day for 3 consecutive days) revealed that CYP1A1 mRNA levels were maximally elevated approximately 10-fold after pyridine treatment for 2 consecutive days, whereas CYP1A2 mRNA levels were maximally elevated approximately 3-fold at 24 hr after treatment. The mRNA levels of glyceraldehyde 3-phosphate dehydrogenase, which served as an internal control, remained constant after 3-MC or pyridine treatment. These results show that expression of CYP1A1 and 1A2 mRNAs is enhanced in renal tissue after exposure to 3-MC or pyridine, and that constitutive expression of CYP1A1 seems to be greater than that of CYP1A2 in renal tissue. PMID- 7493549 TI - Effect of cannabidiol pretreatment on the kinetics of tetrahydrocannabinol metabolites in mouse brain. AB - Tetrahydrocannabinol (THC), a major constituent of marijuana, and several of its metabolites are psychoactive in humans. Cannabidiol (CBD), a nonpsychoactive cannabinoid, inhibits hepatic microsomal THC metabolism and also modulates subjective psychological responses to THC in humans. Treatment of mice with CBD markedly decreased the hepatic microsomal in vitro formation of the major THC metabolites, 6 alpha-OH-THC and 7-OH-THC and increased formation of the minor metabolite 6 beta-OH-THC. THC blood levels were modestly elevated after CBD pretreatment and THC administration, compared with untreated controls, and area under the curve (AUC) of THC increased 50% as a function of decreased clearance. CBD pretreatment modestly increased the Cmax, AUC, or t1/2 of the major THC metabolites in the blood, whereas those kinetic parameters for 6 beta-OH-THC were dramatically increased. Changes in brain concentrations and kinetic parameters of the major THC metabolites did not reflect the relatively modest changes found in blood levels after CBD pretreatment, but exhibited large increases in AUC (7- to 15-fold) and t1/2 (2- to 4-fold), as well as in tmax. Changes in brain concentrations and kinetic parameters for 6 beta-OH-THC reflected the marked changes observed in blood levels after CBD pretreatment. Thus, CBD pretreatment resulted in large increases in AUC and t1/2 of all THC metabolites in brain, with a modest increase in AUC of THC. These changes in THC metabolite brain pharmacokinetics may contribute to the modulation of psychological responses to THC observed after CBD treatment. PMID- 7493550 TI - Disposition of inhaled 1,1,1,2-tetrafluoroethane (HFA134A) in healthy subjects and in patients with chronic airflow limitation. Measurement by 18F-labeling and whole-body gamma-counting. AB - HFA134a (1,1,1,2-tetrafluoroethane) is a nonozone-depleting candidate to replace the chlorofluorocarbons used as propellants in metered-dose inhalers (MDIs) for pharmaceuticals that are widely used in the treatment of respiratory tract disease. As a means for ensuring the safety of such a compound for human use, it is necessary to establish that there is no excessive or unexpected accumulation in the body and in selected regions. A sensitive whole-body gamma-counting technique has been used with 18F-labeled HFA134a to measure the whole-body and regional absorption, distribution, and retention of HFA134a after administration in humans by single-breath inhalation. In seven healthy subjects, labeled HFA134a was rapidly eliminated by ventilation during the first few minutes, with an average of 9.6% of the radioactivity retained in the body at 5 min. This radioactivity cleared with an apparent terminal half-life of 1.5-4.2 hr to leave, on average, < 1% of the administered dose (< 750 micrograms, approximately 0.2 microCi) retained in the body at 5.8 hr. Disposition of radioactivity was independent of the position of label. Thus, there was no evidence of any significant degradative metabolism. On average, only 0.0056% of the administered dose appeared in the urine within the first 2 hr. Later samples contained no significant radioactivity. Inhaled HFA134a first distributed to all regions of the body and then cleared without evident accumulation in any specific region.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7493551 TI - In vivo modification of 3'-phosphoadenosine 5'-phosphosulfate and sulfate by infusion of sodium sulfate, cysteine, and methionine. AB - The importance of tissue sulfate concentrations in regulating 3'-phosphoadenosine 5'-phosphosulfate (PAPS) synthesis is not known. Therefore, this study was conducted to determine the influence of increased availability of inorganic sulfate on steady-state PAPS concentrations in various tissues. To increase tissue sulfate concentrations, 2-16 mmol/kg of sodium sulfate and sulfur containing amino acids (cysteine or methionine) were infused intravenously for 2 hr into pentobarbital-anesthetized rats. Serial blood samples were taken during the infusion and analyzed for sulfate concentrations. After 2 hr of infusion, liver, kidney, and brain were removed for quantification of tissue PAPS and sulfate concentrations. Infusion of sodium sulfate, cysteine, and methionine increased serum and tissue sulfate concentrations in a dose- and time-dependent manner. Serum sulfate concentrations increased from 0.8 to 14 mM during the infusion of sodium sulfate, whereas infusions of cysteine and methionine increased serum sulfate concentrations to 4.8 and 2.0 mM, respectively. Tissue sulfate concentrations also increased during sulfate infusion. Liver sulfate concentration increased from 0.8 to 4.8 mM, kidney concentration increased from 0.6 to 31 mM, and brain concentration increased from 0.1 to 0.6 mM. Similar to the serum sulfate concentrations, sulfate infusion was the most effective in increasing tissue sulfate concentrations, cysteine was intermediate, and methionine the least effective. Although sulfate concentrations in liver, kidney, and brain increased 6-, 50-, and 6-fold by infusing sulfate, respectively; tissue PAPS levels were not altered markedly. Hepatic PAPS concentrations increased significantly (30-35%) only when infused with the higher doses (8 or 16 mmol/kg/2 hr) of sodium sulfate.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7493552 TI - Glutathione-dependent metabolism of trichloroethylene in isolated liver and kidney cells of rats and its role in mitochondrial and cellular toxicity. AB - Metabolism of trichloroethylene (Tri) by the glutathione (GSH) conjugation pathway was studied in hepatocytes, renal cortical cells, and liver subcellular fractions from male Fischer 344 rats. Toxicity of Tri and some of its GSH- and cytochrome P450-dependent metabolites was then studied in isolated hepatocytes, kidney cells, and liver and kidney mitochondria to assess the relative role of these metabolites in toxicity. Tri metabolism to S-(1,2-dichlorovinyl)glutathione (DCVG), a nephrotoxic metabolite of Tri, was demonstrated in both isolated hepatocytes and kidney cells. This suggests that the first Tri bioactivation step that leads to nephrotoxicity in vivo occurs in both liver and kidney. Rates of DCVG formation in liver microsomes and cytosol were similar, although total activity was greater in cytosol. Only S-(1,2-dichlorovinyl)-L-cysteine (DCVC), but not Tri, DCVG, or the other oxidative metabolites examined (trichloroacetate, dichloroacetate, chloral hydrate, trichloroethanol, or oxalic acid) caused acute hepatotoxicity, whereas only DCVC and DCVG produced acute nephrotoxicity in isolated renal cortical cells. Tri and the metabolites examined, except trichloroacetate and DCVG, caused a loss of function of liver and kidney mitochondria. In liver mitochondria, DCVC produced the largest decrease in mitochondrial respiration, whereas Tri, trichloroethanol, and dichloroacetate were somewhat less toxic and chloral hydrate was the least toxic. In kidney mitochondria, in contrast, Tri, trichloroethanol, dichloroacetate, chloral hydrate, and DCVC caused similar decreases in mitochondrial respiration. These results suggest that, whereas both GSH conjugation and cytochrome P450-dependent metabolism of Tri generate mitochondrial toxicants, only the GSH-derived metabolites were cytotoxic. PMID- 7493553 TI - Toxicity and metabolism in mice of 2,6-dithiopurine, a potential chemopreventive agent. AB - 2,6-Dithiopurine (DTP) has been proposed as a possible chemopreventive agent because of its facile reaction with the electrophilic ultimate carcinogen, benzo[a]pyrene diol epoxide, and other reactive electrophiles. Previous studies in mouse skin indicated almost complete inhibition of benzo[a]pyrene diol epoxide induced tumorigenesis by DTP, suggesting the possible utility of this compound as a chemopreventive agent. However, little is known of the metabolism of DTP or of its possible long-term toxicity. Mice were fed diets containing up to 4% DTP in AIN-76A for a period of 7 weeks, and possible toxicity was monitored by weight gain and histopathological examination of all major tissues. No toxicity was observed at any dose of DTP. DTP was found to be a good substrate in vitro for two enzymes known to metabolize 6-mercapto-purine: xanthine oxidase and thiopurine methyltransferase. The in vitro metabolites were 2,6-dithiouric acid and an apparent monomethylated derivative, respectively. In vivo, the major urinary metabolite was 2,6-dithiouric acid, which attained levels as high as 34 mM in the urine of mice receiving the 4% DTP diet. DTP was also excreted unchanged in the feces and urine. DTP, 2,6-dithiouric acid, and an unidentified, relatively nonpolar metabolite were also detected in the serum of experimental animals. Although large interindividual variation in the serum DTP concentration was found, there was a dose-dependent increase in serum DTP as the dietary level of DTP was increased. These results suggest that neither toxicity nor metabolism will severely limit the utility of DTP as a chemopreventive agent. PMID- 7493555 TI - An improved HPLC assay for the assessment of liver slice metabolic viability using 7-ethoxycoumarin. AB - The use of precision-cut liver slices constitutes a new in vitro metabolism technique for the study of coupled phase I and phase II biotransformations. This technique has the advantage of being easily amenable to studies with human tissue. As a means of characterizing the metabolic viability of diverse liver samples, a standard substrate capable of undergoing oxidative and conjugative pathways of metabolism would be desirable. An assay based on 7-ethoxycoumarin was developed whereby the metabolites--7-hydroxycoumarin, 7-hydroxycoumarin sulfate, and 7-hydroxycoumarin glucuronide--could be quantitated using a single HPLC analytical method. This required the synthesis of metabolite standards. 7 Hydroxycoumarin glucuronide was prepared by coupling 7-hydroxycoumarin with methyl 2,3,4-tri-O-acetyl-1-bromo-1-alpha-D- glucopyranuronate, under phase transfer conditions, to give the protected conjugate that was then hydrolyzed to give the glucuronide as the sodium salt. Assignment of configuration at the anomeric center was based on analysis of the simulated 1H and gated 13C NMR spectra, in addition to enzymatic hydrolysis. The sulfate conjugate was prepared by treatment of 7-hydroxycoumarin with Bu4N+ HSO4 /dicyclohexylcarbodiimide/pyridine. 7-Ethoxycoumarin, 7-hydroxycoumarin, and the glucuronide and sulfate conjugates were resolved by HPLC on a C8 Hypersil BDS column using ion-pairing conditions. Incubation of 7-ethoxycoumarin with rat or human liver slices in Krebs-Henseleit buffer resulted in the formation of these metabolites that were readily quantitated in the media with UV detection at 320 nm, using external standard curves. Although the sulfate was seen as the major metabolite in rats, the glucuronide predominated in human tissue. Two different human livers were examined.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7493554 TI - The role of cytochrome P4503A1/2 in the sex-specific sulfoxidation of the hexachlorobutadiene metabolite, N-acetyl-S-(pentachlorobutadienyl)-L-cysteine in rats. AB - Hexachlorobuta-1,3-diene (HCBD) is a selective nephrotoxin and a potent nephrocarcinogen in rodents. Its toxicity and carcinogenicity is based on a multistep bioactivation pathway. Glutathione conjugation seems to be the only bioactivation pathway for HCBD leading to reactive intermediates, which are thought to be responsible for the observed nephrotoxic effects. Recent in vivo studies revealed a novel urinary metabolite in male, but not female, rats after administration of [14C]HCBD. This metabolite was identified as (E)-N-acetyl-S (1,2,3,4,4-pentachlorobutadienyl)-L-cysteine-sulfoxide (N-Ac-PCBC-SO). The objective of this study was to elucidate the enzyme(s) involved in this particular oxidation and to find an explanation for the sex differences in the formation of N-Ac-PCBC-SO. Both cytochrome P450 and flavin-containing monooxygenases (FMO) may catalyze the oxidation of N-Ac-PCBC-SO. The use of various competitive and allosteric inhibitors of cytochrome P450 and FMO (i.e. metyrapone, N-benzylimidazole, thiobenzamide, CO, n-octylamine, and heat inactivation studies) showed that the sulfoxidation of (E)-N-acetyl-S-(1,2,3,4,4 pentachlorobutadienyl)- L-cysteine is catalyzed by cytochrome P450 enzymes. In microsomes from male rats pretreated with pyridine, phenobarbital, and dexamethasone, an increase in the rates of sulfoxide formation was only seen in microsomes from dexamethasone- and phenobarbital-induced animals. Moreover, troleandomycin, a selective chemical inhibitor for enzymes of the cytochrome P4503A family, inhibited sulfoxide formation by > 80%. Correlation of sulfoxide formation with testosterone 6 beta-hydroxylation, a marker of cytochrome P4503A1/2 in the rat, underlined the finding that cytochrome P4503A is the predominant cytochrome P450 responsible for this particular oxidation.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7493557 TI - Placental transfer and fetal disposition of 2',3'-dideoxycytidine and 2',3' dideoxyinosine in the rhesus monkey. PMID- 7493556 TI - Gas-uptake pharmacokinetics and metabolism of 2-chloro-1,1,1,2-tetrafluoroethane (HCFC-124) in the rat, mouse, and hamster. AB - Gas-uptake pharmacokinetics and metabolism of the chlorofluorocarbon replacement 2-chloro-1,1,1,2-tetrafluoroethane (HCFC-124) were investigated in rats, mice, and hamsters. Species differences in the rate of uptake of HCFC-124 and urinary excretion of trifluoroacetic acid were observed. In rats and mice, the uptake of HCFC-124 was described by both saturable and first-order components, whereas in the hamster only first-order uptake was observed. The in vivo metabolic rate constants obtained from computer simulation of the gas-uptake data were: for rats KM = 1.2 mg liter-1 (8.79 mmol liter-1, Vmaxc = 0.35 +/- 0.01 mg kg-1 hr-1 (2.56 +/- 0.01 mmol kg-1 hr-1), and kfc = 1.25 +/- 0.01 hr-1 kg231; for mice-KM = 1.2 mg liter-1 (8.79 mmol liter-1), Vmaxc = 1.78 +/- 0.01 mg kg-1 hr-1 (13.0 +/- 0.007 mmol kg-1 hr-1), and kfc = 4.08 +/- 0.01 hr-1 kg-1; and for hamsters-kfc = 1.47 +/- 0.02 hr-1 kg-1. The production and excretion of trifluoroacetic acid, the major urinary metabolite of HCFC-124, were also simulated in rats and mice, but not in hamsters, by the physiologically based pharmacokinetic model when the in vivo metabolic rate constants obtained in the gas-uptake simulation studies were used. The blood:air partition coefficient of HCFC-124 in the hamster was lower than in the rat or mouse. A low blood:air partition coefficient may limit the pulmonary uptake of volatile chemicals.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7493559 TI - The practical management of thyroid disease in pregnancy. PMID- 7493558 TI - Losartan--a new antihypertensive. PMID- 7493560 TI - Giving erythromycin by mouth. PMID- 7493561 TI - [Preliminary clinical results with intracoronary ultrasound angioplasty]. AB - BASIS AND AIM OF STUDY: Low-frequency, high-intensity ultrasound has been shown, both in vivo and in vitro, selectively to remove arteriosclerotic plaques and thrombi. This study was undertaken to investigate the safety and effectiveness of intracoronary ultrasound angioplasty. PATIENTS AND METHODS: Ultrasound coronary angioplasty (UCA) with highly flexible ultrasound catheters (1.2 mm or 1.7 mm probe tip) was performed in 50 patients (36 men, 14 women; mean age 64.7 [33-79] years) with coronary heart disease involving one (n = 26 or several (n = 24) vessels. Indications for treatment were exercise-induced (n = 35) or unstable (n = 14) angina or acute myocardial infarction (n = 11). Treated lesions were in the anterior interventricular branch (n = 25), circumflex branch of the left coronary artery (n = 3) and right coronary artery (n = 22). 19 vessels were occluded, 24 lesions were partially thrombosed, 19 were calcified. 22 stenoses were longer than 20 mm. According to AHA/ACC criteria, 10 type A, 17 type B1, 6 type B2 and 17 type C lesions were treated. RESULTS: Total ultrasound time was 336 +/- 308 s; mean passing time through the stenosis was 233 +/- 289 (10-556) s. 17 of 19 occlusions were recanalised after 285 +/- 224 s. Percutaneous transluminal angioplasty (PTCA) was subsequently performed in 49 patients. The mean stenosis grade was reduced by UCA from initially 82 +/- 3 to 64 +/- 2% and by subsequent PTCA to 38 +/- 1%. Average flow grade rose from 1.5 to 1.9 after UCA and to 2.8 after PTCA. No vasospasm, atrioventricular block or perforation was caused by UCA. Angiography demonstrated dissection after PTCA in seven patients, treated in two with a stent. Both UCA and PTCA failed to achieve recanalization in one patient with a thrombotic occluded coronary artery after acute myocardial infarction. In all other patients there were no complications. CONCLUSION: The results show UCA to be a safe method for removing high-degree coronary artery stenosis or recanalize thrombotic occlusions. PMID- 7493562 TI - [Factors influencing the prescribing of nootropic drugs. Results of a representative inquiry in Lower Saxony]. AB - AIM OF INVESTIGATION: To discover (1) to what extent patients' wishes and the extent of any abnormality of brain performance influence the frequency with which "nootropic" drugs (those thought to affect brain activity, e.g. piracetam, pyritinol, or improve cerebral circulation, e.g. xanthine derivatives, Ginkgo biloba, secale alkaloids, calcium antagonists) are prescribed; (2) the medical practitioner's expectations of the effectiveness of such medications. METHOD: In a personal interview, 145 family doctors and 14 neurologists in private practice in the Gottingen area of Germany (participation rate: 83.2% of those asked to participate) were questioned about fictitious cases (case 1: mild memory problem with or without expressed wish for medication; case 2: moderate dementia, of Alzheimer or multi-infarct type). The previously arranged interviews, which took place in the doctors' practice rooms, consisted of standardized open questions to the written case reports. RESULTS: Regardless of the wish of the patient and the extent and type of the abnormal brain function about 70% of all participating doctors would prescribe those drugs, even though about 56% had doubts about their effectiveness. About 28% expected a positive effect on brain performance. A nearly equal proportion of doctors would continue an existing drug regimen as would prescribe one. CONCLUSION: The prescription of the named group of drugs is influenced less by medical criteria than by factors which concern doctor-patient relationship. PMID- 7493563 TI - [Diagnosis of pyruvate kinase deficiency in the presence of an elevated reticulocyte count]. AB - HISTORY AND CLINICAL FINDINGS: Three members of one family (mother, now aged 44 years; daughter, now aged 23 years, and son, now aged 19 years) have had chronic haemolytic anaemia since their infancy. All three have had several blood transfusions a year because of haemoglobin (Hb) concentrations as low as 5 g/dl due to intermittent increases in haemolysis. Despite an unclear diagnosis all three had splenectomies: their spleens had been enlarged to about 5-7 cm below the rib margin. After splenectomy their haemoglobin concentration rose to 9-12 g/dl and the previously raised reticulocyte count of 150-250/1000 rose further to 400-700/1000. None of the patients had symptoms other than a slight decrease in exercise tolerance. They were admitted to hospital for further evaluation. Physical examination was unremarkable except for slight jaundice and well healed splenectomy scar. TESTS: All three patients had normochromic haemolytic anaemia (Hb 10-12 g/dl), bilirubin 2-3 mg/dl, lactate dehydrogenase activity 400-600 U/l). The reticulocyte count was raised to 420-690/1000. In all of them pyruvate kinase (PK) activity was normal or slightly increased if related to Hb concentration. Reticulocytes from healthy persons were isolated by density gradient centrifugation so that samples could be prepared with different numbers of reticulocytes: PK activity was found to be dependent on the sample's reticulocyte count. In the three patients PK deficiency could, therefore, only be diagnosed by taking into account the raised reticulocyte count. CONCLUSION: As reticulocytes have a higher PK activity than erythrocytes the reticulocyte count must be taken into account when assessing the significance of a given value of this enzyme's activity. PMID- 7493564 TI - [Value of chemotherapy in multi-modal treatment of non-small-cell bronchial carcinoma]. PMID- 7493565 TI - [Hereditary non-polyposis colon carcinoma. Syndrome of the cancer family--a neglected disease]. PMID- 7493566 TI - ["Privatization" of a radiologic hospital department. Decision of the German Federal Social Court of March 15, 1995]. PMID- 7493567 TI - [Discrepancy between peripheral thyroid hormone values and TSH level]. PMID- 7493568 TI - [Vocal register changes during use of oral contraceptives or thyroid hormones]. PMID- 7493569 TI - [Percutaneous extracorporal circulation in cardiogenic shock caused by a mixed poisoning with metildigoxin, nifedipine and indapamide]. PMID- 7493570 TI - [Povidone-iodine eyedrops for the prevention of newborn conjunctivitis]. PMID- 7493571 TI - [Endoscopic drainage of pancreatic pseudocysts]. AB - AIM OF STUDY: To compare retrospectively the endoscopic treatment of pancreatic pseudocysts by retrograde pancreatic drainage (ERPD), cystogastrostomy (ECG) or cystoduodenostomy (ECD). PATIENTS AND METHODS: Between 1987 and 1995 pancreatic pseudocysts were successfully treated endoscopically in 30 patients (13 women, 17 men; mean age 49 [18-89] years): ERPD was accomplished through papillary insertion of 5-7 F prostheses into the cysts or pancreatic duct in 9 patients; ECG achieved drainage of cysts in the body or tail of the pancreas in 15 patients, and ECD for cysts in the head of the pancreas in three. Combined ERPD and ECG was necessary in three patients. RESULTS: Significant pain relief was achieved in all patients, accompanied by increased appetite and weight. Occlusion or dislocation of the prosthesis required renewed endoscopic drainage in three patients. The prostheses were removed, 2-12 months after implantation, when pain relief had occurred and clinical as well as radiological findings had become normal. Recurrence 10-22 months after removal of the drain was observed in four patients, of whom two had a successful second drainage procedure. Surgical intervention became necessary in three patients because of inadequate drainage or (one case) bleeding. CONCLUSION: These data indicate that endoscopic drainage of pancreatic pseudocysts gives good results with a low complication rate and no deaths. The procedure is thus an alternative to the at present prevailing operative treatment. PMID- 7493572 TI - [Effect of lowered cholesterol on the course of coronary heart disease. An analysis of the results of controlled, angiographically documented intervention studies]. AB - AIM: To find out from published reports whether the tendency of coronary heart disease (CHD) to progress can be retarded by lowering total and LDL cholesterol concentrations. METHODS: After a data-base search a meta-analysis was undertaken of all those randomized, controlled and angiographically documented studies which contained informations about the effect of cholesterol reduction on the course of CHD over a period of at least 2 years. A total of 12 studies covering 3781 patients met the stated criteria. RESULTS: The different lipid-lowering measures (usually drugs) achieved a statistically significant reduction of the number of patients with progression of the coronary angiographic findings and a significant increase in the number of those with actual regression. The number of coronary incidents, such as myocardial infarction, unstable angina, sudden cardiac death, necessary aortocoronary bypass operations or percutaneous transluminal coronary angioplasty was lower by 34% in the groups with measures to influence cholesterol metabolism than in the untreated groups. In the studies in which coronary arteriograms were evaluated quantitatively there was an annual increase in the mean degree of stenosis of 1.01% in the untreated and 0.37% in the treated groups. CONCLUSIONS: The difference in the increase of the mean degree of stenosis is probably not only important because of the resulting haemodynamic changes in myocardial perfusion. The quantitatively only slightly increased trend towards progression in the untreated groups can be interpreted as a marker for an active process of atherogenesis with a tendency of plaques to break off and of plaque thromboses. PMID- 7493573 TI - [Successful valvuloplasty of a calcified, high-grade pulmonary valve stenosis with removal of a right-left shunt at the atrial level]. AB - HISTORY AND CLINICAL FINDINGS: A 68-year-old woman, with a cardiac murmur known since childhood and suffering from increasing dyspnoea for several years, was admitted to hospital after echocardiography had suggested pulmonary valve stenosis and an atrial septal defect. While her general condition was satisfactory, she had marked central cyanosis with clubbed fingers and moderate bilateral oedema of the lower legs. A rough, diamond-shaped systolic murmur was heard, loudest over the left 2nd intercostal space. In addition to a cardiac defect with right to left shunt, primary pulmonary disease with cor pulmonale was considered in the differential diagnosis. EXAMINATIONS: Haemoglobin content was 25.1 g/dl, haematocrit 72.4%, red cell count 7.44 x 10(6). Arterial oxygen partial pressure was 40 mm Hg, arterial oxygen saturation 81.8%, Echocardiography further revealed right ventricular enlargement, marked tricuspid regurgitation and a stenosed calcified pulmonary valve. At right heart catheterization a right to left interatrial shunt was calculated at 47% of systemic flow and a systolic pressure gradient between right ventricle and pulmonary artery of 131 mm Hg was measured. TREATMENT AND COURSE: Haemoglobin content was lowered to 19.4 g/dl after bloodletting. Balloon pulmonary valvuloplasty reduced the transvalvular systolic gradient to 31 mm Hg. The further course was without complication: 4 months later the patient had only grade II (NYHA) dyspnoea, the oxygen saturation was 91.3%. CONCLUSION: Severe pulmonary valve stenosis with right to left interatrial shunt can be successfully treated by balloon dilatation even in elderly patients. PMID- 7493574 TI - [The significance of so-called "high-intensity transient signals" (HITS) during transcranial Doppler sonography]. PMID- 7493575 TI - [Methods in medicine: dynamic magnetic resonance tomography]. PMID- 7493576 TI - [Kidney transplantation in patients with diseases of the gastrointestinal tract and the liver]. PMID- 7493577 TI - [Humidifiers]. PMID- 7493578 TI - [Diagnosis of immune defects in Candida esophagitis]. PMID- 7493579 TI - [Spontaneous course of inguinal hernias]. PMID- 7493581 TI - [Differential diagnosis of mononucleosis]. PMID- 7493580 TI - [Diagnostic value of cytokine determination in serum and plasma]. PMID- 7493582 TI - [Epidemiology of echinococcosis in Bavaria]. PMID- 7493583 TI - [Long-term survival with HIV infection. Virological-immunological studies of pathogenesis]. PMID- 7493584 TI - Neonatal cerebral arterial flow velocity waveforms in term infants with and without metabolic acidosis at delivery. AB - To define the effects of acid base status at delivery on neonatal cerebral artery flow velocity waveform patterns obtained using Doppler ultrasound during the first week of life, a longitudinal comparative study of neonates born at term with and without evidence of metabolic acidosis in the umbilical artery was undertaken. Eighty-two appropriate for gestational age infants delivered after uncomplicated pregnancies with non-acidotic umbilical artery blood gases and in whom no neonatal complications were noted were studied to establish reference values of neonatal cerebral arterial vascular resistance index (RI) in normal term infants during the first week of life. A further 189 infants were grouped according to the presence and severity of metabolic acidosis at delivery, and also the presence of high risk features in the antenatal period. In the normal non-acidotic infants, over the first 24 h of life, there was a significant fall in the cerebral arterial resistance index (RI) in all the vessels examined, after which a steady state value was attained with no significant changes in vascular resistance index being noted during the remainder of the study period. The fall in RI between 12 and 24 h of age was consistent in all study groups. Infants with metabolic acidosis at delivery had blood flow patterns compatible with decreased resistance to flow in both anterior and middle cerebral arteries which persisted throughout the first week of life. This reduction in cerebral vascular resistance was most marked in those infants with severe metabolic acidosis. The majority of severely acidotic infants had a benign clinical outcome in the first week of life and all infants had normal cerebral ultrasound scans during the neonatal period. These findings suggest that metabolic acidosis at birth is associated with changes in neonatal cerebral arterial vascular resistance during the first week of life, and in the presence of benign clinical course the significance of this observation with regard to neurodevelopmental outcome requires evaluation. PMID- 7493585 TI - Perinatal thermogenesis. AB - The rapid initiation of thermogenesis is crucial for the survival of newborn infants. At birth the fetus must adapt to cooling, increased oxygenation and separation from the placenta. An experimental approach in the chronically instrumental fetal sheep of 'simulated birth in utero' allowed the evaluation of each of these stimuli sequentially. Cooling stimulated shivering, cardiovascular and endocrine responses but not nonshivering thermogenesis (NST). Ventilation of the cooled fetus with oxygen caused only modest NST which was not altered by an infusion of triiodothyronine. Occluding the umbilical cord was followed by a rapid substantial rise in NST which was maintained until the placental circulation was re-established. Thus the placenta is secreting factors into the fetal circulation which inhibit the ability of the brown adipose tissue to respond to either hormonal or neural stimuli. Placental prostaglandin E2 and probably adenosine are tonic inhibitors of thermogenesis in utero. Effective thermogenesis after birth requires the combination of separation from the placental inhibitors of lipolysis, increased oxygenation from breathing and the stimulation of cutaneous cold receptors. PMID- 7493586 TI - Reduction of masseter muscle activity in bottle-fed babies. AB - Our previous studies suggested that there are significant differences in the growth of the jaw and in muscle activity between breast- and bottle-fed infants. To confirm these differences quantitatively, myoelectric activities of the masseter muscles of bottle-fed babies were studied. Twelve bottle-fed babies, as well as 12 breast-fed babies as a control group, were examined electromyographically during bottle or breast feeds. The duration time of sucking bursts, interval time, cycle time, 0-to-peak amplitude, integrated amplitude of bursts, and integrated amplitude/duration time were measured and the number of bursts over 30 microV was counted. All differences of means were significant by t test. The masseter muscle activity in bottle-fed babies is significantly reduced. Our results are contrary to previous papers in which almost the same sucking actions in both breast and bottle feeding were reported. The reason why previous researchers thought that the sucking patterns in breast- and bottle-fed babies are essentially the same is considered, and the implications of the differences for dental health are discussed. PMID- 7493587 TI - Skin blood flow oscillations respond more effectively to rhythmic thermal stimulation than to continuous or periodic breathing in newborn infants. AB - Effect of continuous (n = 30) and periodic (n = 5) breathing on forehead skin blood flow (SBF) oscillations was studied in healthy 3-day-old preterm and term infants. Femoral SBF was measured in 10 of the term infants during continuous breathing. Respiratory and SBF signals were analysed by power spectrum during the control period. The signal analysis was repeated during rhythmic thermal stimulation which was applied to the skin of each infant in order to synchronise spontaneous SBF oscillations. During the control period, the forehead and femoral SBF of all the studied infants oscillated at frequencies corresponding to the frequency of regular continuous breathing. External thermal stimulation decreased these high-frequency oscillations of forehead SBF in both preterm and term infants. Oscillations in femoral SBF of term infants remained unchanged. During periodic breathing, the spontaneous forehead SBF oscillation was synchronised to the frequency of respiratory amplitude variation. Despite periodic breathing, thermal stimulation was able to resynchronise the SBF oscillation to the stimulation frequency in 4/5 infants. Forehead SBF oscillates at the frequency of both continuous and periodic breathing in preterm as well as in term infants on postnatal day 3. Rhythmic thermal stimulation synchronises SBF and affects SBF more effectively than continuous or periodic breathing. These results suggest functional neural control of cutaneous vasculature at birth. PMID- 7493588 TI - Prevention of intraventricular hemorrhage in preterm infants. AB - Prematurely born infants with intraventricular hemorrhage (IVH) suffer significant morbidity and mortality, particularly those infants with high grade hemorrhage. The more premature infants have a higher incidence, experiencing more severe IVH. Early onset IVH is also likely to be severe and to progress to a higher grade. The etiology of intraventricular hemorrhages is clearly multifactorial, with differing sets of risk factors for early onset and later occurring hemorrhage. Prevention requires multilayered strategies, both prenatal and postnatal. These strategies are discussed in detail, highlighting unresolved controversies. Certain recommendations for prevention can be made. These include efforts to prevent preterm delivery, transfer of high risk mothers to tertiary care centers and antenatal maternal steroid use. Postnatally, the importance of optimal resuscitation and neonatal care practices is stressed, particularly those which minimize cerebral blood flow fluctuation. Postnatal indomethacin use should be considered in most infants. Further investigation of other strategies is necessary, including multicenter randomized trials to further evaluate antenatal pharmacologic agents, as well as the relative efficacy of different modes of delivery. The different risk factors for early onset versus later onset IVH must be more clearly delineated. Most importantly, any strategy must include sustained neurodevelopmental followup. PMID- 7493589 TI - Microbial characteristics and severity of coagulase-negative staphylococcal septicaemia in premature neonates. AB - A pilot study was undertaken to assess the predictive value of characteristics possessed by coagulase-negative staphylococci in determining clinically important infection in premature neonates. No significant association was demonstrated between organism properties and clinical indices of infection. The routine use of simple measures of microbial characteristics in the assessment of neonatal septicaemia is not warranted. PMID- 7493590 TI - Comparison between the essential fatty acid status of preterm and full-term infants, measured in umbilical vessel walls. AB - The essential fatty acid (EFA) composition of umbilical vessel walls is increasingly being studied as a longer-term reflection of the fetal EFA status. We evaluated the EFA content of umbilical artery and vein vessel walls in 43 preterm infants and compared it with that of 43 full-term cord vessels. In addition, relations among cord vessel wall fatty acid composition, gestational age (GA) at birth, and anthropometric parameters at birth (weight, head circumference, and length) were explored in the preterm infants. Generally, n-6 and n-3 EFA levels were lower, while levels of EFA deficiency markers were higher in preterm than in term cords, both in the walls of the draining arteries and the supplying vein. In preterm cords, significant correlations were observed between GA at birth and levels of n-6 and n-3 EFAs (positive) and EFA deficiency markers (negative). Birth weight showed significant (P < or = 0.01), positive correlations with n-6 and n-3 EFA levels in the cord artery walls of preterm infants, all after correction for GA at birth. In conclusion, substantial differences between the EFA profiles of preterm and full-term cord vessel walls indicate a lower biochemical EFA status of the preterm than of the term fetus. This lower preterm EFA status might be a reflection of a physiologically lower EFA demand for growth and development of the preterm fetus than of the term fetus. PMID- 7493591 TI - A powerful bacterial world. AB - Bacteria (prokaryotes) were the sole form of life on earth for some two billion years--roughly half its history. During this time they evolved into a giant, global superorganism and developed a remarkable mechanism for the creation and exchange of genetic material. Apart from its intrinsic interest, this has practical significance, for example in the development of multiple resistance to antibiotics of pathogenic bacteria such as those of tuberculosis. Eukaryotes, with nucleated cells, may have developed from a permanent symbiosis of three or more prokaryotes. PMID- 7493592 TI - Biomechanics of snoring. AB - A large proportion of the population either snores or suffers the snoring of others. Recent advances with the use of fibre-optic endoscopes have enabled surgeons to observe the inside of the pharynx while a patient is asleep and snoring. In this article we look at the underlying structure of the upper airway and explain, with the use of simple mechanical models, the aerodynamic events occurring inside the upper airway during snoring. PMID- 7493593 TI - Inter- and intra-individual differences in semantic priming among young and older adults. AB - The influences of inter- and intra-individual differences on context effects for young and older adults were examined. Results of a semantic priming task revealed two subsets of older adults who were markedly different in terms of response latency, consistency of responding, and magnitude of priming. Examination of individual response distributions indicated that (a) age differences in priming were localized to trials representing subjects' slowest responses and were eliminated for their fastest responses and (b) inconsistently responding older individuals contributed disproportionately to age differences in priming. The results are interpreted in terms of an interactive-compensatory model in which the ability to sustain attention is related to both priming and word recognition. It is concluded that the failure to examine individual differences may be contributing to the discrepant results in the literature. PMID- 7493594 TI - Working memory resource allocation by young, middle-aged, and old adults. AB - Three hypotheses regarding the locus of age differences in working memory were examined in an experiment in which young, middle-aged, and old adults remembered the names of persons standing in line while also performing mental addition. Instructions as to the relative importance of these two tasks were manipulated. The results are highly compatible with the hypothesis of an age-related deficit in a single pool of resources that can be allocated to different tasks. They do not support hypotheses of deficits in one or more pools used separately for processing and storage functions. Middle-aged adults performed as well as young adults and better than old adults. PMID- 7493595 TI - Free recall and recognition of slowly and rapidly presented words in very old age: a community-based study. AB - This study addressed the effects of study time on episodic recall and recognition of words in a community-based sample of healthy older adults ranging from 75 to 96 years of age (N = 221). Results indicated a slight but reliable age-related deterioration of free-recall performance that was attributable to age deficits in secondary memory. The size of the age-related impairment in recognition was reduced relative to that in recall. As well, for all age groups, recall and recognition were higher when items were slowly as opposed to rapidly presented, indicating proficient utilization of study time in very old age. Finally, multiple regression analyses indicated that, although a variety of demographic (i.e., age and education), psychometric (i.e., Mini-Mental State Exam [MMSE] and Block Design scores), and biological (i.e., thyroid-stimulating hormone) variables were related to free-recall performance, only Block Design and MMSE scores made independent contributions to recognition performance. PMID- 7493597 TI - Personality change in dementia of the Alzheimer's type: relations to caregiver personality and burden. AB - Using the NEO Personality Inventory (NEO-PI), we evaluated caregivers' perceptions of personality prior to symptom onset and current personality in 36 individuals with a clinical diagnosis of dementia of the Alzheimer's type (DAT). Caregivers also completed the self form of the NEO-PI and an index of objective and subjective burden. Personality change in DAT was consistent with previous reports of increased neuroticism, decreased extraversion, and decreased conscientiousness, with smaller decreases in openness and agreeableness. Significant relationships were found among perceived present patient personality, caregiver personality, and caregiver burden. Regression analyses indicated that present patient conscientiousness and caregiver neuroticism were the best predictors of both objective and subjective burden, and these variables were found to contribute independently to caregivers' reported level of burden. To the extent that caregivers perceive specific behaviors and interferences negatively, cognitive and supportive interventions are likely to prove extremely beneficial. PMID- 7493596 TI - Life events as correlates of burden in spouse caregivers of persons with Alzheimer's disease. AB - The first goal of this study was to examine the occurrence and appraised distress of 20 stressful life events in 175 spouse caregivers (mean age = 68.6) of persons with Alzheimer's disease and 92 age- and gender-matched controls (mean age = 68.5). The second goal was to determine whether, in the caregivers, life stressors were related to burden after controlling for caregiver age, gender, satisfaction with social supports, and care-recipient cognitive and activities of daily living functioning. Caregivers and controls had similar occurrences and appraisals for all but 3 events, which occurred more frequently in caregivers (i.e., "changes in your spouse's health," "changes in your relationship with your spouse," and "changes in the way you friends and family treat you (social relations)"). Only "changes in finances" was appraised as more distressing by caregivers. Six events explained burden in the regressions: "victim of crime," "age discrimination," "serious family arguments (not including spouse)," "change in caregiver's health," "trouble with Medicaid/Medicare or Social Security," and "move to retirement home (associated with less burden)." Age discrimination was related to burden only for the caregivers of the most impaired care recipients. Also, caregivers who experienced health problems and who had been victims of crimes were the most burdened. Caregivers are not immune to problems experienced by older adults, and these problems may exacerbate caregiver burden. Clinicians should consider the overall context of a caregiver's life when evaluating burden. PMID- 7493598 TI - Colonic microvascular integrity in acute endotoxaemia: interactions between constitutive nitric oxide and 5-lipoxygenase products. AB - Administration of the nitric oxide synthase inhibitor, NG-nitro-L-arginine methyl ester (5 mg/kg s.c.) provoked acute microvascular injury (assessed by the leakage of radiolabelled human serum albumin) in the rat colon within 1 h, when administered concurrently with endotoxin (Escherichia coli lipopolysaccharide, 3 mg/kg i.v.). Pretreatment with the selective inhibitor of 5-lipoxygenase, BW A137C (N-[4-benzyloxybenzyl] acetohydroxamic acid; 1-20 mg/kg s.c., 15 min before endotoxin) attenuated such damage in a dose-dependent manner. These findings suggest a balance between protective constitutive nitric oxide and the detrimental actions of 5-lipoxygenase products in the maintenance of vascular integrity in the early stage of sepsis. PMID- 7493599 TI - The suppression by lipopolysaccharide of cytochrome P450-dependent renal vasodilation in the rat is mediated by nitric oxide. AB - The isolated perfused kidney of the rat was used to examine the hypothesis that lipopolysaccharide-induced nitric oxide (NO) production inhibits cytochrome P450 dependent vasodilation. The vasodilator responses to arachidonic acid and bradykinin were examined as the response to arachidonic acid is wholly dependent, and that to bradykinin partly dependent on cytochrome P450 metabolism. In endotoxin-treated rats, the vasodilator response to arachidonic acid was inhibited, and those to bradykinin and acetylcholine were enhanced. Following treatment with phenobarbitone, the inducer of certain isoforms of cytochrome P450 enzymes, the vasodilator effects of all three agonists, especially that of arachidonic acid, were amplified. Lipopolysaccharide inhibited the effect of phenobarbitone on the vasodilator effect of arachidonic acid and bradykinin but enhanced that of acetylcholine. The effect of lipopolysaccharide was antagonized by haemoglobin, a NO antagonist, and N omega-nitro-L-arginine, an inhibitor of NO synthase, suggesting that the inhibitory effect of lipopolysaccharide on arachidonic acid- and bradykinin-induced vasodilation was mediated by NO/NO synthase. N omega-Nitro-L-arginine enhanced vasodilation induced by arachidonic acid while that induced by bradykinin or acetylcholine was reduced, implying that endogenous NO inhibits vasodilator cytochrome P450 metabolites in the rat kidney. Pretreatment with dexamethasone, an inhibitor of inducible NO synthase, resulted in inhibition of the lipopolysaccharide modulation of arachidonic acid-induced vasodilation, suggesting that the inducible NO synthase is the target of the inhibitory effect of lipopolysaccharide. The inhibitory effect of lipopolysaccharide was mimicked by nitroprusside, the L-arginine-independent NO donor, and by L-arginine, the biosynthetic precursor of NO. The effect of L arginine, but not of nitroprusside, was antagonized by N omega-nitro-L-arginine, suggesting a specific role for NO synthase in the inhibitory effect of lipopolysaccharide in the inhibition of cytochrome P450-dependent vasodilation in the rat kidney. PMID- 7493600 TI - Effect of lidocaine on alpha 1-adrenoceptors in cultured neonatal rat cardiocytes. AB - The effect of lidocaine on alpha 1-adrenoceptors in cultured neonatal rat ventricular cardiocytes was studied by binding assay. When the cells were cultured in the presence of lidocaine, the binding of (+/-)-beta-([125I]iodo-4 hydroxyphenyl)-ethyl-aminomethyl-tetralone to the cells increased in a concentration- and time-dependent manner. The effect was due to an increase in maximum binding but not due to a change in the dissociation constant for the ligand. The increased number of alpha 1-adrenoceptors returned to the control level after culturing the cells without lidocaine. The stimulating effect on the number of alpha 1-adrenoceptors of lidocaine was also observed in the presence of norepinephrine. Other class I antiarrhythmic drugs such as procainamide. disopyramide, mexiletine and flecainide increased the number of alpha 1 adrenoceptors in the cells. These results suggest that cardiac responsiveness mediated by alpha 1-adrenoceptors is increased by class I antiarrhythmic drugs. PMID- 7493601 TI - Behavioural response to SKF 38393 and quinpirole following chronic antidepressant treatment. AB - The effects of chronic administration of antidepressant drugs (21-22 days s.c. via osmotic mini-pumps) on the behavioural responses of male Sprague-Dawley rats to (-)-quinpirole hydrochloride (0.05 mg kg-1 s.c., 5 min) and (+/-)-SKF 38393 hydrochloride (10 mg kg-1 s.c., 5 min) were investigated. Desipramine hydrochloride (10 mg kg-1 per day), phenelzine sulphate (10 mg kg-1 per day) and clorgyline hydrochloride (1 mg kg-1 per day) attenuated the suppression of locomotor activity induced by quinpirole, a dopamine D2-like receptor agonist, while clomipramine hydrochloride (10 mg kg-1 per day) was without effect. Yawning elicited by quinpirole was absent in phenelzine- and clorgyline-treated rats, but unaffected in rats treated chronically with desipramine and clomipramine. SKF 38393, a dopamine D1-like receptor agonist, significantly increased locomotor activity and time spent grooming in control animals. There were no significant effects of antidepressants on the behavioural responses to SKF 38393. PMID- 7493602 TI - Somatotropin and somatostatin effects on vitamin K-dependent plasma coagulation factors. AB - The effects of somatotropin (0.2 mg/kg body mass) and somatostatin (0.1 mg/kg body mass) on plasma coagulation factors II, VII, IX, X and some general indexes of hemocoagulation were examined. Hormones were injected subcutaneously in male Wistar rats on 3 consecutive days. Boehringer Mannheim tests and Schnitger and Gross coagulometer were used for clotting factor determination. Somatotropin caused significantly decreased activity of factors II, VII and X (P < 0.001) and IX (P < 0.05). Somatostatin alone, as well as somatotropin after somatostatin pretreatment considerably increased the activity of factors II, VII and X (P < 0.001), while factor IX was non-significantly suppressed. It is concluded that somatotropin and somatostatin are possible regulators of biosynthesis of vitamin K-dependent plasma coagulation factors. Somatotropin depresses the activity of factors II, VII, IX and X and causes hypocoagulability, while somatostatin not only prevents the inhibiting effect on factors II, VII and X, but also increases their activity and causes hypercoagulability. PMID- 7493603 TI - Motor actions of 7-OH-DPAT in normal and reserpine-treated mice suggest involvement of both dopamine D2 and D3 receptors. AB - In non-habituated mice, 7-hydroxy-N,N-di-n-propylaminotetralin (7-OH-DPAT, 0.04 10 mg/kg s.c.) potently and rapidly suppressed species-typical behaviours and induced frozen postures, with only occasional evidence of weak behavioural stimulation occurring at 5-10 mg/kg. This inhibitory effect was reversed by the dopamine D1 receptor agonist 2,3,4,5-tetrahydro-7,8-di-hydroxy-1-phenyl-1H-3 benzazepine hydrochloride (SKF 38393, 10 mg/kg i.p.). 7-OH-DPAT (3-10 mg/kg) did not reinstate locomotion in 4 h habituated mice, either when administered alone or in conjunction with a threshold dose of SKF 38393 (3 mg/kg). By contrast, 7-OH DPAT (0.2-10 mg/kg) dose-dependently reversed the akinesia of 24 h reserpine treated mice. This response was blocked by the dopamine D2 receptor antagonist raclopride (10 mg/kg i.p.), but not by the dopamine D1 receptor antagonist (R) (+)-8-chloro-2,3,4,5-tetrahydro-3-methyl-5-phenyl-1H-3- benzazepine-7-ol hemimaleate (SCH 23390, 0.05 mg/kg i.p.), and was potentiated synergistically by coinjection of SKF 38393 (3 mg/kg). These and earlier data suggest the motor inhibitory effects of 7-OH-DPAT (low doses) in normal animals are mediated by dopamine autoreceptors (D2 and/or D3), whilst its motor stimulant actions in normal (high doses) and in dopamine-depleted, supersensitive animals, are mediated by dopamine D2 receptors. PMID- 7493604 TI - Pharmacological studies of allergic cough in the guinea pig. AB - The pharmacological mechanisms of allergic cough in the guinea pig were studied. Actively sensitized guinea pigs were exposed to aerosols of antigen to elicit coughing. In separate experiments, naive guinea pigs were exposed to aerosols of capsaicin to elicit coughing. Both allergic and capsaicin-induced cough were inhibited by loratadine (0.3-10 mg kg-1 p.o.) and chlorpheniramine (0.1-3.0 mg kg 1 p.o.). Neither cimetidine (10 mg kg-1 s.c.), nor thioperamide (3-10 mg kg-1 s.c.), inhibited allergic or capsaicin-induced cough. Codeine (3-30 mg kg-1 p.o.), salbutamol (0.003-3.0 mg kg-1 s.c.) and ipratropium (0.03-1.0 mg kg-1 s.c.) inhibited both allergic and capsaicin-induced cough. Hexamethonium (10 and 30 mg kg-1 s.c.) inhibited allergic, but not capsaicin-induced cough. Allergic and capsaicin-induced cough were unaffected by phenidone (5.0 and 10.0 mg kg-1 s.c.). Indomethacin (5.0 and 10.0 mg kg-1 s.c.) had no effect on allergic cough but slightly inhibited capsaicin-induced cough. We conclude that allergic and capsaicin-induced cough are modulated by histamine H1 receptor and cholinergic mechanisms. Histamine H2 or histamine H3 receptor mechanisms, and lipoxygenase and cyclooxygenase products of arachidonic acid metabolism do not influence allergic and capsaicin-induced cough. Ganglionic mechanisms play a minor role in the production of allergic cough and no role in capsaicin-induced cough. PMID- 7493605 TI - The activation gate of cardiac Na+ channel modulates voltage- and pH-dependent unbinding of disopyramide. AB - To assess the drug unbinding process from receptor sites in cardiac Na+ channels, we examined the recovery kinetics of disopyramide-blocked Na+ current (INa) in isolated guinea-pig ventricular myocytes using the whole-cell variation of the patch-clamp technique. In the presence of disopyramide (20 microM), the time course of INa recovery from use-dependent block (unbinding) was described by a double exponential function. Although the time constant for the fast phase (tau f) of recovery was unchanged at different membrane voltages, the slow phase (tau s) increased with hyperpolarizing membrane potential: 4.4 +/- 0.2 s at a holding potential of -90 mV and 6.4 +/- 0.3 s at -140 mV (n = 10, P < 0.01). The slow time constant of INa recovery was also increased by acidification. These findings suggest that disopyramide molecules can escape from the receptor site through the hydrophobic pathway after deprotonation, because slowing of recovery from use dependent block by acidification is caused by a decreased deprotonation rate of receptor-bound drug molecules. In addition to the hydrophobic escape, the roles of the fast inactivation gate and activation gate (m-gate) were evaluated during the recovery process. After inhibition of the fast inactivation process of INa by pretreatment with chloramine-T (2 mM), the fast phase of recovery from use dependent block by disopyramide was abolished.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7493606 TI - Caffeine and carbachol act on common Ca2+ stores to release Ca2+ in guinea-pig ileal smooth muscle. AB - To characterize intracellular Ca2+ stores, the Ca(2+)-releasing effects of caffeine, carbachol and inositol 1,4,5-trisphosphate (IP3) were compared by measuring the drug-induced tension development in beta-escin-skinned longitudinal smooth muscle of guinea-pig ileum. Caffeine (20 mM), carbachol (10 or 100 microM) or IP3 (40 microM), applied after loading Ca2+ within intracellular stores, produced a transient rise in tension in a Ca(2+)-free solution. This change in tension occurred in response to release of Ca2+ from the stores. The effect of either caffeine or carbachol was markedly reduced or abolished after preceding application of the other drug. IP3 was without effect when applied subsequently to caffeine. The effects of carbachol and IP3 were abolished after combined treatment with ryanodine (30 microM) and caffeine (20 mM) which causes functional removal of caffeine-releasable Ca2+ stores, but not after combined treatment with ryanodine (30 microM) and carbachol (10 microM). The results suggest that caffeine, carbachol and IP3 all act on common Ca2+ stores to release Ca2+. PMID- 7493607 TI - The 5-HT2 receptor antagonist, pelanserin, inhibits alpha 1-adrenoceptor-mediated vasoconstriction in vitro. AB - The antagonism by pelanserin (2,4(1H,3H)-quinazolinedione,3-[3-(4-phenyl-1- piperazinyl)-propyl]-HCl), a potent 5-HT2 receptor antagonist, of alpha 1 adrenoceptor-mediated contractions of endothelium-denuded carotid, aorta, mesenteric and caudal arteries of both normotensive Wistar-Kyoto (WKY) and spontaneously hypertensive (SHR) rats was investigated. The selective alpha 1 adrenoceptor agonist methoxamine elicited concentration-dependent contractions in all four arterial rings, an effect which was competitively antagonized by pelanserin. pA2 values for pelanserin were in the 7.67-8.11 range when evaluated against the alpha 1-adrenoceptor agonist in arteries from normotensive or hypertensive rats. These data support the conclusion that pelanserin displays alpha 1-adrenoceptor blocking properties. The ability of the 5-HT2 receptor antagonist pelanserin to additionally block alpha 1-adrenoceptor-mediated constriction in different vessels of WKY and SHR may potentially contribute to its blood pressure lowering effects. PMID- 7493608 TI - Role of oxygen-derived metabolites in the rat gastric mucosal injury induced by nitric oxide donors. AB - Local intra-arterial infusion of high doses of the nitric oxide (NO) donor, nitroprusside (10-40 micrograms kg-1 min-1 for 15 min) induced dose-dependent haemorrhagic injury to the rat gastric mucosa and reduced systemic arterial blood pressure, whereas intragastric nitroprusside (10-50 mg ml-1), which caused similar falls in blood pressure, failed to induce such injury. The mucosal damage induced by nitroprusside was reduced by local concurrent infusion of superoxide dismutase (500-4000 i.u. kg-1). Local superoxide dismutase also abolished the mucosal injury induced by local infusion of the NO donor, S-nitroso-N-acetyl penicillamine (40 micrograms kg-1 min-1), but not that induced by local infusion of endothelin-1 (5 pmol kg-1 min-1) indicating specific actions. Intravenous infusion of the iron chelator and peroxyl scavenger, desferrioxamine (0.25-1 mg kg-1 min-1) or the hydroxyl radical scavenger, dimethylthiourea (20 mg kg-1 min 1) also reduced the mucosal damage induced by the local administration of the NO donors, but not that induced by endothelin-1. These findings implicate the involvement of superoxide and possibly other oxygen-derived free radicals in the injurious actions of high levels of nitric oxide generated from NO donors, and may reflect a role of the cytotoxic peroxynitrite moiety. PMID- 7493610 TI - Dopamine receptor agonist potencies for inhibition of cell firing correlate with dopamine D3 receptor binding affinities. AB - The potencies for in vivo inhibition of substantia nigra pars compacta dopamine single cell firing were determined for apomorphine, BHT 920, N-0923, (+/-)-7 hydroxy-dipropylaminotetralin (7-OH-DPAT), (+)-3-(3-hydroxyphenyl)-N propylpiperidine (3-PPP), pramipexole, quinelorane, quinpirole, RU 24926, U 86170, and U-91356. Significant correlation was obtained between the potencies of these 11 highly efficacious dopamine receptor agonists and the in vitro binding affinities at dopamine D3 receptors, but not at dopamine D2L receptors. These results support a functional role for the dopamine D3 receptor subtype in the autoreceptor-mediated regulation of dopamine cell activity, while a role for dopamine D2 receptors awaits further analysis. In addition, the results demonstrate the limitations of using currently available dopamine receptor agonists to delineate relative in vivo roles for the dopamine D2 and D3 receptor subtypes. PMID- 7493611 TI - Receptor binding and functional evidence suggest that postsynaptic alpha 2 adrenoceptors in rat brain are of the alpha 2D subtype. AB - This study has determined the subtype(s) of postsynaptic alpha 2-adrenoceptors in rat brain. This question has been addressed by using two separate approaches, i.e. ligand displacement of [3H]2-(2-methoxy)-1,4-benzodioxan-2-yl)-2-imidazoline ([3H]RX 821002) from membranes prepared from rat cortex after noradrenergic denervation and, secondly, by antagonism of clonidine-induced mydriasis. After rats had been lesioned using N-(2-chloroethyl)-N-ethyl-2-bromobenzylamine (DSP-4; 100 mg/kg i.p., 30 min after zimeldine 10 mg/kg i.p.), noradrenaline was undetectable in the cortex 3 days later. Displacement of [3H]RX 821002 with a range of agonists and antagonists which distinguish between the known alpha 2 adrenoceptor subtypes (alpha 2A-2D) yielded pKi values which correlated very well with reported values for the alpha 2D-adrenoceptor (r = 0.929; P < 0.001), but not the alpha 2A (r = 0.450; P = 0.192), alpha 2B (r = 0.280, P = 0.434) or alpha 2C (r = 0.283; P = 0.460) subtypes. Similarly, the potencies of various alpha 2 adrenoceptor antagonists to inhibit clonidine (0.03 mg/kg i.p.)-induced mydriasis in conscious rats correlated strongly with their pKi values for alpha 2D adrenoceptors (r = 0.899; P = 0.015) but not alpha 2A-(r = 0.369; P = 0.472), alpha 2B-(r = -0.224; P = 0.670) or alpha 2C-adrenoceptors (r = 0.253; P = 0.584). These data are, therefore, consistent and argue strongly that postsynaptic alpha 2-adrenoceptors in the rat cortex and Edinger-Westphal nucleus are of the alpha 2D subtype. PMID- 7493609 TI - NMDA receptors are involved at the ventrolateral nucleus tractus solitarii for termination of inspiration. AB - The purpose of the present study was to determine whether blockade of excitatory amino acid receptors at the ventrolateral nucleus of the tractus solitarius would influence respiratory activity. This was done by microinjecting excitatory amino acid receptor antagonists into the ventrolateral nucleus of the tractus solitarius of alpha-chloralose-anesthetized animals while monitoring respiratory activity using a Fleisch pneumotachograph and arterial blood pressure and heart rate. Bilateral microinjection of the NMDA receptor antagonist, 3-[(R) carboxypiperazin-4-yl]-propyl-1- phosphomic acid (CPP), 5.62 nmol per side, produced an increase in inspiratory duration (+4 +/- 1.6 s, n = 8) which progressed to an apneustic pattern of breathing. Similar results were obtained with CPP microinjected into the ventrolateral nucleus of the tractus solitarius of three vagotomized animals. Bilateral microinjection of a second NMDA receptor antagonist, 2-amino-7-phosphono-heptanoic acid (AP7), 562 nmol per side, produced qualitatively similar effects on respiration as seen with CPP. In contrast, blockade of non-NMDA receptors with 6-cyano-7-nitroquinoxaline-2,3-dione (CNXQ), 0.125 nmol per side, had very little effect on respiration. Activation of NMDA receptors at the ventrolateral nucleus of the tractus solitarius with bilateral microinjection of NMDA, 39 pmol, produced a large increase in expiratory duration (+11 +/- 3 s, n = 8), and apnea during the expiratory phase of the respiratory cycle in half of the animals studied. Similar results were obtained with D,L alpha-amino-3-hydroxy-5-methyl-4-isoxazol-proprionate (AMPA). These results indicate that an endogenous excitatory amino acid released at the ventrolateral nucleus of the tractus solitarius and acting at the NMDA receptor, plays a significant role in respiratory timing. PMID- 7493613 TI - Bicuculline enhances the late GABAB receptor-mediated paired-pulse inhibition observed in rat hippocampal slices. AB - The inhibition of CA1 pyramidal neurones in rat hippocampal slices was studied using extracellular recordings of population spike potential responses to paired orthodromic stimulation. Variation of the interpulse interval allowed the separation of an early phase of inhibition (interpulse interval 5-20 ms), blocked by the GABAA receptor antagonist bicuculline (1 microM; n = 11), and a late phase (interpulse interval 200-400 ms) blocked by the GABAB receptor antagonist phaclofen (1 mM; n = 5) but enhanced by bicuculline (n = 11). Similar enhancement was not observed when conditioning response amplitudes were increased by increasing the stimulus strength, rather than bicuculline. Orthodromic stimulation leads to synaptic excitation of both pyramidal neurones and inhibitory interneurones, and may also lead to activation of inhibitory inputs onto interneurones. Bicuculline could prevent inhibition of the interneurones, and hence enhance the late, GABAB receptor-mediated inhibition. Conversely, the therapeutic administration of benzodiazepines would be postulated to enhance the inhibition of inhibitory interneurones, leading to an iatrogenic decrease in GABAB receptor-mediated inhibition. PMID- 7493612 TI - The selective angiotensin AT1 receptor antagonist LR-B/081 potently inhibits drinking induced by central injection of angiotensin II in rats. AB - LR-B/081, methyl-2-[[4-butyl-2-methyl-6-oxo-5-[[2'-(1H-tetrazol-5-yl) [1,1' biphenyl]-4-yl] methyl]-1(6H)-pyrimidinyl] methyl]-3-thiophenecarboxylate, is a recently developed nonpeptide antagonist selective for angiotensin AT1 receptors. The drug has been reported to be an insurmountable angiotensin AT1 receptor antagonist endowed with long-lasting antihypertensive activity. A large body of evidence indicates that angiotensin AT1 receptors mediate the dipsogenic action of angiotensin II in the central nervous system. The present study evaluated the ability of LR-B/081, in comparison with losartan and with its active metabolite EXP3174, to inhibit drinking induced by central injection of angiotensin II in water-sated rats. LR-B/081, in the dose range of 10-1000 pmol/rat, dose dependently inhibited the drinking response to angiotensin II, 10 pmol/rat. The ID50 of LR-B/081 was 25.9 pmol/rat, while that of losartan and EXP3174 was 357 and 3.9 pmol/rat, respectively. Therefore LR-B/081 was about 7 times less potent than EXP3174, but about 14 times more potent than the parent molecule losartan. LR-B/081 altered neither carbachol-induced water intake, nor 15% fat milk intake in rats, suggesting that its effect on angiotensin II-induced drinking is a behaviourally selective effect. These findings show that LR-B/081 potently inhibits central angiotensin AT1 receptors involved in behaviourally selective effect. These findings show that LR-B/081 potently inhibits central angiotensin AT1 receptors involved in the control of body fluid homeostasis and suggest that this drug might be an interesting pharmacological tool to further investigate the role of the central renin-angiotensin system in physiological or pathological conditions. PMID- 7493614 TI - Short- and long-term changes in striatal and extrastriatal dopamine uptake sites in the MPTP-treated common marmoset. AB - The 'short-term' (15-30 days) and 'long-term' (18-42 months) effects of the systemic administration of 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP) on [3H]mazindol binding to dopamine uptake sites was investigated in the common marmoset. In the 'short-term' MPTP-treated group, [3H]mazindol binding was reduced in the caudate-putamen (by -82 to -98% with respect to controls), substantia nigra pars compacta (-71 to -84%), ventral tegmental area (-72%) and nucleus accumbens (-54%). [3H]Mazindol binding in the globus pallidus, frontal cortex and substantia nigra pars reticulata was much lower and was unaffected by MPTP treatment. In the 'long-term' MPTP-treated group [3H]mazindol binding was still greatly reduced in the substantia nigra pars compacta (by -76 to -89%), ventral tegmental area (-71%) and most of the caudate-putamen (-69 to -98%), although the reduction in [3H]mazindol binding in the nucleus accumbens (-27%) and rostroventral caudate nucleus (-69%) was less than in the 'short-term' MPTP treated group. The motor deficits induced by MPTP treatment in the common marmoset are largely reversible with increasing survival times (Ueki et al., 1989, Neuropharmacology 28, 1089). In the present study, the apparent 'recovery' in [3H]mazindol binding in the rostroventral caudate nucleus and nucleus accumbens may indicate regeneration of dopamine neurone terminals in these regions and this may contribute to the behavioural recovery seen in this primate model of Parkinson's disease. PMID- 7493615 TI - Modulation of acetylcholine, capsaicin and substance P effects by histamine H3 receptors in isolated perfused rabbit lungs. AB - The modulatory role of histamine H3 receptors in pulmonary oedema induced by acetylcholine, capsaicin and by exogenous substance P was investigated in isolated, ventilated rabbit lungs. Endothelial permeability was evaluated by measuring the capillary filtration coefficient (Kf,c). Acetylcholine (10(-8) to 10(-4) M), substance P (10(-10) to 10(-6) M), capsaicin (10(-4) M) and 5 hydroxytryptamine (5-HT) (10(-4) M) induced an increase in the Kf,c. Carboperamide, a novel histamine H3 receptor antagonist, induced a significant leftward shift of the concentration-response curve to acetylcholine and also enhanced the effect of capsaicin on the Kf,c, while it had no significant effect on the response to substance P and 5-HT. Imetit, a new histamine H3 receptor agonist, strongly inhibited the effects of acetylcholine and capsaicin. Imetit also strongly protected the lung against substance P effects but did not prevent the 5-HT-induced increase in the Kf,c. Carboperamide completely blocked the inhibitory effect of Imetit on the acetylcholine response. (R)-alpha Methylhistamine, an other histamine H3 receptor agonist, had the same protective effect against acetylcholine response as Imetit. We conclude that histamine H3 receptors could protect the lung against acetylcholine- and capsaicin-induced oedema via a prejunctional modulatory effect on the C-fibres. However, since the response to exogenous substance P was also inhibited by histamine H3 receptor stimulation, the presence of such receptors at a postsynaptic level, probably on mast cells, was also suggested. PMID- 7493616 TI - Spinal delta 2-, but not delta 1-, mu-, or kappa-opioid receptors are involved in the tail-flick inhibition induced by beta-endorphin from nucleus raphe obscurus in the pentobarbital-anesthetized rat. AB - The antinociception induced by beta-endorphin given supraspinally has been previously demonstrated to be mediated by the release of [Met5]enkephalin acting on delta-opioid receptors in the spinal cord. The present study was designed to determine what type of opioid receptors in the spinal cord is involved in beta endorphin-induced antinociception in the rat. Antinociception was induced by beta endorphin (0.6 nmol) given into nucleus raphe obscurus and was assessed by the tail-flick test in pentobarbital-anesthesized rats. Naltriben (0.6-6.0 nmol), a selective delta 2-opioid receptor antagonist, given intrathecally dose dependently attenuated beta-endorphin-induced inhibition of the tail-flick response. On the other hand, 7-benzylidene naltrexone (2.1-64.3 nmol), CTOP (D Phe-Cys-Tyr-D-Trp-Orn-Thr-Pen-Thr-NH2, 0.09-2.8 nmol), or nor-binaltorphimine (1.4-40.8 nmol), selective delta 1-, mu-, and kappa-opioid receptor antagonists, respectively, did not block beta-endorphin-induced antinociception. The results of present study in rats are consistent with previous experiments in mice indicating that spinal delta 2-, but not delta 1-, mu- or kappa-opioid receptors are involved in beta-endorphin-induced inhibition of the tail-flick response. PMID- 7493618 TI - N-nitro-L-arginine methyl ester blocks the decompensatory phase of acute hypovolaemia in conscious rabbits by a brainstem mechanism. AB - Graded caval occlusion in conscious rabbits caused a biphasic response. Phase I was characterized by a fall in conductance so that arterial pressure was maintained. When cardiac output had fallen to 71 +/- 4% of its baseline level, phase II supervened. During phase II, conductance rose abruptly and arterial pressure fell to a life threatening level (< 40 mm Hg). When administered into the fourth ventricle, the nitric oxide synthase inhibitor N-nitro-L-arginine methyl ester prevented the onset of phase II. The mean threshold dose for this effect was 4 mumol (range: 0.4-11). When administered intravenously, a dose of 275 mumol N-nitro-L-arginine methyl ester prevented the onset of phase II in only one out of six rabbits. It is concluded that a central brainstem nitrergic mechanism is involved in the onset of the decompensatory phase II of the haemodynamic response to hypovolaemia. PMID- 7493617 TI - The in vitro pharmacological characterization of naloxone benzoylhydrazone. AB - On the basis of its in vivo activity and binding affinity, naloxone benzoylhydrazone has been characterized as a kappa 3-opioid receptor agonist and a mu-opioid receptor antagonist. This paper continues its pharmacological characterization with the help of isolated tissue preparations. Naloxone benzoylhydrazone was found to have partial agonist activity in the guinea pig ileum longitudinal muscle/myenteric plexus preparation. As an antagonist, naloxone benzoylhydrazone is similar to naloxone, with pA2 values of 8.8, 7.8, and 7.8 for mu-, delta-, and kappa 1-opioid receptors, respectively. Its agonist activity in the guinea pig ileum preparation was not influenced by beta funaltrexamine treatment but was reversed by the selective kappa-opioid receptor antagonist nor-binaltorphimine and by the irreversible kappa 1-opioid receptor blocker UPHIT (1S,2S)-trans-2-isothiocyanato-4,5-dichloro-N-methyl-N-[2-(1- pyrrolidinyl)-cyclohexyl] benzeneacetamide. The presence of kappa 3-opioid receptors could not be demonstrated by [3H]naloxone benzoylhydrazone binding in the guinea pig ileum longitudinal muscle/myenteric plexus preparation. From these studies it is concluded that the partial agonist activity of naloxone benzoylhydrazone in this bioassay is probably due to the activation of the kappa 1-opioid receptors. PMID- 7493619 TI - The KATP blocker sodium 5-hydroxydecanoate does not abolish preconditioning in isolated rat hearts. AB - Blockers of ATP-sensitive K+ channels (KATP) abolish preconditioning in several species. Glyburide does not abolish preconditioning in rat hearts, but this may be due to a loss of its activity during ischemia. We determined the effect of a KATP blocker, which is more active during ischemia (sodium 5-hydroxydecanoate, 5 HD), on preconditioning in isolated rat hearts. Rat hearts were subjected to 4 periods of 5 min global ischemia followed by 30 min of global ischemia and reperfusion. Preconditioning significantly enhanced post-ischemic recovery of function and reduced lactate dehydrogenase (LDH) release vs. sham. 5-HD (100 microM) did not abolish preconditioning. Cromakalim (20 microM) was protective in this ischemic model and this was abolished by 5-HD. This is further evidence that KATP opening is not the mechanism of preconditioning in rats. PMID- 7493620 TI - Antinociceptive effect of a kappa-opioid receptor agonist that minimally crosses the blood-brain barrier (ICI 204448) in a rat model of mononeuropathy. AB - The antinociceptive effect of intraplantar (i.pl.) ICI 204448 ((R,S)-N-[2-(N methyl-3,4-dichloro-phenylacetamido)-2-(3-carbox yph enyl)- ethyl]pyrrolidine hydrochloride)) (20, 30, 40 and 50 micrograms), a kappa-opioid receptor agonist which has limited access to the central nervous system, was studied in a well established rat model of peripheral mononeuropathy produced by moderate constriction of the sciatic nerve. Vocalization thresholds to paw pressure were used as a nociceptive test. On the injected nerve-injured paw, ICI 204448 at 20 and 30 micrograms had no significant effect, but higher doses (40 micrograms) produced a significant antinociceptive effect, which plateaued at 50 micrograms. By contrast, no antinociceptive effect was observed on the contralateral paw. The effect of ICI 204448 (40 micrograms) was significantly antagonised by the specific kappa-opioid receptor antagonist nor-binaltorphimine (20 and 30 micrograms), when co-injected in the nerve-injured paw. PMID- 7493623 TI - Alternative splicing of fibronectin--many different proteins but few different functions. PMID- 7493621 TI - Evidence for the involvement of nitric oxide in the antinociceptive effect of ketorolac. AB - The involvement of nitric oxide in the antinociception produced by ketorolac was assessed using the pain-induced functional impairment model in the rat: 800 micrograms of NG-nitro-L-arginine methyl ester, an inhibitor of nitric oxide synthesis, or saline was injected intra-articularly in a hind limb joint previously injured with uric acid. Animals then received ketorolac, dipyrone or no drug. Ketorolac and dipyrone produced a significant antinociceptive effect which was reduced by pretreatment with NG-nitro-L-arginine methyl ester, but not with saline. It is concluded that the antinociceptive effect of both drugs involves the local participation of nitric oxide. PMID- 7493622 TI - BW373U86, a delta-opioid receptor agonist, reverses bradykinin-induced thermal allodynia in rhesus monkeys. AB - The synthetic delta-opioid receptor agonist BW373U86 (0.18-0.56 mg/kg s.c.) was studied in rhesus monkeys with a warm-water, tail-withdrawal assay, designed to detect bradykinin (0.1 microgram) and prostaglandin E2 (5-15.8 micrograms) induced thermal allodynia. BW373U86 dose-dependently reversed bradykinin allodynia, but was ineffective against prostaglandin E2 allodynia. The BW373U86 dose-effect curve was shifted to the right by the delta-opioid receptor-selective antagonist naltrindole (1.0 mg/kg) but not by the mu-opioid receptor-selective antagonist quadazocine (0.1 mg/kg). The present findings add to the conditions in which delta-opioid receptor-mediated behavioral effects have been detected in primates, and suggest that delta-opioid agonists may be of therapeutic interest in the treatment of some types of hyperalgesic conditions. PMID- 7493624 TI - Intracellular routing and release of caseins and growth hormone produced into milk from transgenic mice. AB - Growth hormone (GH) secretion, in mammary tissue from transgenic mice, containing a chimeric gene composed of the regulatory region of whey acidic protein gene and the structural region of GH gene, was compared to casein secretion. GH was expressed in milk and for a small percentage (1:1000) in blood as revealed by SDS polyacrylamide gel electrophoresis and radioimmunoassay. As attested by immunofluorescence and immunogold electron microscopy, caseins and GH followed the same secretory pathway. However, contrary to caseins, which are essentially in micellar form, GH was detected in a nonaggregated form in secretory vesicles and in the lumen of the acini. Newly synthesized caseins and GH were carried simultaneously, mainly to the lumen of the acini, but also to the base of the cell. Secretion of newly synthesized proteins was increased by prolactin (PRL). As shown by immunoblotting, the proportion of GH versus other proteins, secreted in the presence of PRL was not modified, suggesting that GH secretion is subjected to the same hormonal regulation by PRL as other milk proteins. These results show that, in lactating mammary epithelial cells from transgenic mice, a recombinant GH and the caseins are carried simultaneously to the lumen and suggest that secretion of both proteins is increased by PRL during the same time course. Transport of these newly synthesized proteins occurs also to the base of the cell. PMID- 7493625 TI - Changes of mitochondrial mass in the hemopoietic stem cell line FDCP-mix after treatment with etoposide: a correlative study by multiparameter flow cytometry and confocal and electron microscopy. AB - FDCP-Mix, a pluripotent murine hemopoietic stem cell line which undergoes typical internucleosomal cleavage of DNA when induced to apoptosis by either drugs or withdrawal of growth factor (interleukin-3) was studied after treatment with the topoisomerase II inhibitor etoposide (0.5-4 microM). An increase in autolytic activity was the major early morphological change within the cytoplasm, with mitochondria as the main target for autolytic digestion. Despite this macroautophagy, thin sections showed a high number of mitochondria, suggesting mitochondrial proliferation as a result of drug treatment. This observation of an increase in the number of mitochondria was confirmed by flow cytometric studies of mitochondrial overall mass. Multiparameter flow cytometry of cells double stained with propidium iodide and nonyl-acridine orange gave an accurate assay for mitochondrial mass in relation to cell cycle stages. The increase in mitochondrial mass was found in all cell cycle stages. The results suggest a drug induced proliferation of mitochondria separate from the processes involved in the doubling of mitochondrial mass during the cell cycle and a decline of mitochondria in the later stages of apoptosis. PMID- 7493626 TI - Involvement of nitric oxide in permeability alteration and F-actin redistribution induced by phorbol myristate acetate in endothelial cells. AB - We have previously reported that phorbol myristate acetate (PMA) caused a decrease in endothelial permeability during the first 1 to 1.5 h of exposure and thereafter an increase for up to 6 h. This permeability alteration was correlated with a time-dependent redistribution of F-actin, i.e., an increase in dense peripheral bands was observed during the first hour of PMA incubation and a disruption of the bands after 6 h. In the present study, we found that this PMA induced alteration of permeability is L-arginine dependent, since the low permeability prevailed for up to 6 h when extracellular L-arginine was available. Moreover, we noted that administration of N-nitro-L-arginine methylester (L-NAME) to PMA-treated cells caused a direct increase in permeability. The redistribution of F-actin induced by PMA was also L-arginine dependent, since the number of dense peripheral bands continued to increase for up to 6 h when extracellular L arginine was available, and these bands were directly disrupted when L-NAME was added. These results suggest that the tight contact between PMA-treated endothelial cells is maintained by a redistribution of F-actin elicited by the endogenous production of nitric oxide. PMID- 7493628 TI - Analysis by confocal microscopy of the behavior of heat shock protein 70 within the nucleus and of a nuclear matrix polypeptide during prolonged heat shock response in HeLa cells. AB - By means of confocal laser scanning microscopy and indirect fluorescence experiments we have examined the behavior of heat-shock protein 70 (HSP70) within the nucleus as well as of a nuclear matrix protein (M(r) = 125 kDa) during a prolonged heat-shock response (up to 24 h at 42 degrees C) in HeLa cells. In control cells HSP70 was mainly located in the cytoplasm. The protein translocated within the nucleus upon cell exposure to hyperthermia. The fluorescent pattern revealed by monoclonal antibody to HSP70 exhibited several changes during the 24 h-long incubation. The nuclear matrix protein showed changes in its location that were evident as early as 1 h after initiation of heat shock. After 7 h of treatment, the protein regained its original distribution. However, in the late stages of the hyperthermic treatment (17-24 h) the fluorescent pattern due to 125 kDa protein changed again and its original distribution was never observed again. These results show that HSP70 changes its localization within the nucleus conceivably because it is involved in solubilizing aggregated polypeptides present in different nuclear regions. Our data also strengthen the contention that proteins of the insoluble nucleoskeleton are involved in nuclear structure changes that occur during heat-shock response. PMID- 7493629 TI - Motility of vinculin-deficient F9 embryonic carcinoma cells analyzed by video, laser confocal, and reflection interference contrast microscopy. AB - We have studied the motility of wild-type F9 and vinculin-deficient (5.51) mouse embryonal carcinoma cells. F9 cells extended filopodia at a rate of 61 ( +/- 18) nm/s over a distance of 3.18 (+/- 0.29) microns. In contrast, 5.51 cells exhibited filopodia which extended at a similar speed of 57 (+/- 17) nm/s but over a longer distance of 5.10 (+/- 2.14) microns. Cell-substratum contact areas of both cell types were examined by reflection interference contrast microscopy. Wild-type F9 cells had distinct close contacts (dark gray areas) at the cell periphery, whereas 5.51 cells had only a few light gray pinpoint contacts with the substrate. Confocal microscopy showed alpha-actinin to be localized along actin stress fibers in wild-type cells, and in 5.51 cells stress fibers were absent and alpha-actinin was associated with F-actin in the filopodia. beta 1 integrin, talin, and paxillin were concentrated in focal contacts in wild-type cells, but in 5.51 cells beta 1-integrin and talin were in patches under the plasma membrane and paxillin was diffusely distributed in the cytoplasm. We conclude that changes in cell shape and motility of 5.51 compared to wild-type F9 cells are due to the absence of vinculin even though there may be functions of other focal adhesion complex proteins, e.g., talin, linking the actin cytoskeleton to the plasma membrane. PMID- 7493627 TI - Identification of a transposon-related RNA down-regulated by retinoic acid in embryonal carcinoma and embryonic stem cells. AB - The differential display polymerase chain reaction was employed to identify changes in mRNA expression during retinoic acid-induced differentiation in embryonal carcinoma PCC4.aza1R cells. In this study, we report on one cDNA, EC1, that was identified by this method. EC-1 encodes a 0.6-kb mRNA that is present in PCC4.aza1R cells and down-regulated by retinoic acid. Sequence analysis revealed that EC-1 exhibits a 47% identity with the early transposon RNA ETn and does not contain a long open reading frame. EC-1 mRNA expression was reduced by 50% after 24 h of treatment with 10 nM retinoic acid and was undetectable after 48 h. Down regulation of EC-1 mRNA was observed at retinoic acid concentrations as low as 0.1 nM. EC-1 was found to be expressed in several other embryonal carcinoma cell lines as well as in embryonic stem cells but was undetectable in differentiated cell types obtained after RA treatment. Northern blot analysis using RNA from multiple mouse tissues demonstrated that the expression of EC-1 is restricted to the testis. Treatment of PCC4.aza1R cells with an RAR-selective agonist also repressed the expression of EC-1 mRNA while treatment with an RXR-selective agonist reduced EC-1 expression slightly. The RAR alpha-specific antagonist Ro 41 5253 had little effect on the down-regulation of EC-1 by retinoic acid. Our observations indicate that the repression of EC-1 is associated with the induction of differentiation in embryonal carcinoma and embryonic stem cells and involves an RAR-activated signaling pathway. PMID- 7493630 TI - 50-kb chromatin fragmentation in the absence of apoptosis. AB - Treatment with ionic detergents of nuclei isolated from various continuously growing cell lines generally yields chromatin samples of high viscosity. Extensive treatment with nuclease-free proteinase K or pronase solubilized the viscous lysates with > 90% of the DNA migrating at approximately 50 kb. Freshly prepared human peripheral blood T cells also yield a substantial fraction of their DNA in an approximately 50- to 100-kb band. The cleavage sites may coincide with a class of DNase I-hypersensitive regions, since digestion of chromatin by DNase I at approximately 10 U/ml, without protease, also yields fragments of preferentially approximately 50-kb size. Occasionally, the oligonucleosomal ladder was also detected together with high molecular weight degradation products. Remarkably, all of these fragmentation patterns were seen in healthy, resting or proliferating cells, i.e., in the absence of apoptosis. Tritiated thymidine incorporation could be readily detected in the approximately 50-kb DNA fragments. The effect of an apoptotic intracellular milieu on the integrity of isolated chromatin is apparently imitated by the extensive protease treatment used in our DNA isolation protocol. PMID- 7493631 TI - Detection of nucleotide excision repair incisions in human fibroblasts by immunostaining for PCNA. AB - During nucleotide excision repair, damaged DNA is incised on both sides of a lesion and an oligomer containing the damage is excised and replaced by repair DNA synthesis. The latter step is accomplished in vitro by proteins that include the DNA polymerase accessory factor PCNA, which binds to DNA ends to initiate repair synthesis. An increased association of PCNA with nuclei occurs after UV irradiation of nonreplicating DNA in normal human fibroblasts, probably following incision of damaged DNA. This property was used to detect the catalysis of nucleotide excision repair incisions in damaged DNA in vivo, by immunostaining of quiescent human fibroblasts with the widely available PC10 antibody. We summarize here a comprehensive survey of PCNA immunostaining in repair-defective xeroderma pigmentosum (XP) cells in comparison to normal cells. XP-A and XP-G cells were completely defective in staining for PCNA 30 min after UV irradiation. This strongly suggests that XPA and XPG proteins are absolutely required in cells before any incisions can be formed in damaged DNA. XP-B, XP-C, XP-D, and XP-F cells showed an intermediate level of staining for PCNA after UV irradiation, indicative of partial incision capacity in those cells. UV-irradiated XP-E and XP V cells showed normal PCNA immunostaining levels, consistent with evidence that the corresponding factors are not essential for the incision step of repair. The results provide further evidence for the involvement of PCNA in the repair process in vivo and give an alternative to traditional approaches for measurement of nucleotide excision repair capability. PMID- 7493632 TI - Distribution of caldesmon and of the acidic isoform of calponin in cultured cerebellar neurons and in different regions of the rat brain: an immunofluorescence and confocal microscopy study. AB - Caldesmon and calponin are two F-actin-binding and calcium-calmodulin-dependent proteins. In smooth muscle and nonmuscle cells both proteins are localized on actin filaments. Using one- or two-dimensional gel electrophoresis followed by the Western blot technique, and by immunofluorescence studies, we have given evidence that calponin is also present in rat and pig brain. In the present study, for the first time, we demonstrate caldesmon- and calponin-specific immunoreactivities in cerebellar cultured neurons. In the rat central nervous system these antibodies mainly stain neuronal cell bodies and dendrites. By confocal analysis we observed that calponin and caldesmon are located in the actomyosin domain although the total actin and myosin were not saturated. In many cases it is clear that these two proteins are adjacent rather than superimposed in the same domain of the cell. These results are compatible with the functional role of caldesmon and calponin in the regulation of the actomyosin activity as described by others and suggest that they are part of the contractile apparatus of neural cells. PMID- 7493633 TI - Cell type- and differentiation stage-dependent expression of PML domains in rat, detected by monoclonal antibody HIS55. AB - Using mouse monoclonal antibody (mAb) HIS55, we identified a nuclear antigen (ag) that exhibited a staining pattern of discrete foci. Such foci could be detected in cells of many mammalian species. These nuclear foci were not associated with nuclear membrane, nucleoli, or mitotic chromosomes. In isolated rat liver nuclei, HeLa cells, and normal human and rat lymph nodes, staining of HIS55 colocalized with that of 5E10. 5E10 recognizes PML nuclear domains, multimolecular complexes of unknown function containing the product of PML gene and at least two other components. HIS55 foci were expressed widely in many tissues but the expression level varied in a cell type-specific manner, with the number of HIS55 nuclear foci ranging from 0 (as in neurons) to over 100 (as in megakaryocytes) and the size ranging from fine (as in cortical thymocytes) to very large (as in urethra epithelium). HIS55 ag expression level also varied among cells of the same lineage, as observed in embryonic development of rat and in the hemopoietic system of adult rat. The expression level of HIS55 foci roughly correlated with the overall rate of protein synthesis of cells, supporting a role of PML domains as transcription regulatory units. The expression of HIS55 foci, however, did not correlate with the growth index of cell populations. Our observations on normal tissues agreed with the hypothesis that PML domain expression is regulated by external, possibly site-dependent factors. We further supported this by demonstrating that PML domains in rat ventral prostate epithelia were upregulated upon castration. PMID- 7493634 TI - Attachment of Pneumocystis carinii to primary cultures of rat alveolar epithelial cells. AB - Pneumocystis carinii (PC) is an exclusively extracellular pathogen which causes pneumonia in immunocompromised individuals. Histologic studies have demonstrated that PC organisms attach preferentially to type I alveolar epithelial cells and rarely bind to type II cells. Previous reports have demonstrated that cultured type II cells develop a type I cell-like phenotype and express type I cell surface antigens. The current study examines the attachment of PC organisms to isolated rat type II alveolar epithelial cells as a function of time in culture. PC attachment to isolated type II cells increased as the type II cells differentiated in culture from 2.3 +/- 1.2% on Day 2 to 18.4 +/- 2.7% by Day 8. Previous studies have indicated a role for fibronectin (Fn) and Fn receptors as mediators of PC attachment. Addition of anti-Fn antibodies decreased attachment of PC to Day 8 type II cells from 19.4 +/- 2.5% to 9.4 +/- 1.9% (P < 0.01). Addition of antibodies to the alpha v and alpha 5 integrin subunits resulted in significant decreases in PC attachment to Day 8 type II cells. Examination of expression of alpha v and alpha 5 integrins on Day 2 and Day 8 type II cells demonstrated increased expression of both alpha v and alpha 5 integrin subunits on Day 8 type II cells. Overall these data indicate that attachment of PC to isolated rat type II cells increases as the cells differentiate into a type I cell-like phenotype in vitro and correlates with increased expression of Fn binding integrins on the cell surface of the cultured type II cells. PMID- 7493635 TI - Localization of the Bcl-2 protein to the outer mitochondrial membrane by electron microscopy. AB - Incubation with the Bcl-2 antibody before fixation of tissues allowed good localization of the antigenic determinant. We showed that the Bcl-2 gene product in centroblastic-centrocytic lymphoma is mainly localized on the outer mitochondrial membrane and, to a lesser degree, on the nuclear envelope. No significant staining was found in other cytoplasmic domains. Careful examination also revealed that gold particles did not recognize an integral membrane epitope, but an antigenic determinant localized at a short distance from the cytoplasmic side of the membrane itself. This observation suggests that, by interacting with other cytoplasmic proteins, Bcl-2 plays some role in the cytoplasmic machinery involved in the regulation of programmed cell death. PMID- 7493636 TI - Regulation of osteoblast gene expression in intratypic osteosarcoma hybrid cells. AB - Intratypic osteosarcoma hybrids were constructed by fusing the human osteoblast like osteosarcoma SaOS-2 with the rat osteoblast-like osteosarcoma UMR-106. Both of these osteosarcomas express liver/bone/kidney alkaline phosphatase (ALPL), but only the UMR-106 cell line expresses osteopontin (OPN), a gene expressed during later stages of osteoblast differentiation. Analysis of osteoblast gene expression in these hybrids demonstrated that ALPL continued to be expressed; however, OPN steady-state mRNA levels were dramatically reduced in four hybrids. Quantitative measurements indicated that OPN steady-state mRNA levels were extinguished by a factor of 20- to 1000-fold. Since SaOS-2 chromosomes are preferentially lost from these hybrids, subclones of extinguished hybrids were isolated that reexpressed OPN mRNA at levels similar to the UMR-106 parental line. These data indicate that trans-acting negative regulatory factors, expressed from the SaOS-2 genome, are responsible for OPN extinction. This report provides the first demonstration of the negative regulation of OPN gene expression and also provides additional evidence that extinction plays a role in the regulation of osteoblast gene expression. PMID- 7493637 TI - Retinoic acid specifically increases nuclear PKC alpha and stimulates AP-1 transcriptional activity in B16 mouse melanoma cells. AB - B16 melanoma cells differentiate upon treatment with retinoic acid (RA). This differentiation process is accompanied by an increase of protein kinase C alpha (PKC alpha) mRNA and protein levels. Overexpression of PKC alpha in these cells results in a more differentiated phenotype, suggesting the importance of this protein in the control of differentiation by RA. The purpose of the study reported here was to determine the subcellular distribution of the RA-induced PKC alpha, whether the RA-induced increase in PKC alpha protein levels was accompanied by an increase in in situ enzyme activity, and whether RA altered AP 1 transcriptional activity. We found that RA treatment increased PKC alpha protein levels in all subcellular compartments examined, but it also induced a selective enrichment in nuclear-associated PKC alpha levels. Treating cells with an active phorbol ester induced translocation of PKC alpha to membrane fractions, but had no effect on nuclear PKC alpha levels. RA also increased PKC enzymatic activity in intact cells as determined by phosphorylation of the PKC-specific endogenous substrate MARCKS. However, while RA induced a five- to eightfold increase in total cellular PKC alpha protein levels, it only increased MARCKS phosphorylation by twofold. In light of the increase in in situ PKC enzyme activity and the enrichment of nuclear PKC alpha, we determined whether AP-1 activity might be increased in RA-treated cells. Use of luciferase reporter gene constructs with or without AP-1 elements transfected into B16 cells indicated that RA induced a four- to fivefold increase in AP-1 transcriptional activity. These results suggest a hypothesis whereby RA-induced nuclear PKC alpha might lead to increased AP-1 activity and show that RA-induced growth inhibition and differentiation are not always accompanied by an inhibition of AP-1 activity as has been proposed by other investigators. PMID- 7493638 TI - Differential modulation of integrin receptors and extracellular matrix laminin by transforming growth factor-beta 1 in rat alveolar epithelial cells. AB - The transforming growth factors-beta (TGFs-beta) family of genes plays important roles in cell growth and differentiation in many cell types. TGF beta modulates the synthesis and accumulation of extracellular matrix (ECM) components and the expression of cell surface receptors for ECM components. TGF beta is increased in alveolar lining fluid during inflammatory reactions of the lung and has been identified in alveolar epithelial cells of developing lungs and hyperplastic type II cells during repair. However, little is known about how TGF beta may regulate expression of extracellular matrix proteins and ECM receptors in lung alveolar epithelial cells. Laminin, a major glycoprotein component of epithelial basement membrane, is synthesized and secreted by alveolar epithelial cells. To study the effects of TGF beta on modulation of laminin and its integrin receptors alpha 6 beta 1 and alpha 3 beta 1 in lung alveolar epithelial cells, a rat alveolar type II cell-derived cell line, LM5, was incubated with TGF beta 1 (0-100 pg/ml) in serum-free medium for 0-16 h. We examined the expression of integrin subunits and laminin beta 2 chain (s-laminin) mRNAs and protein expression. By Northern blot analysis, TGF beta 1 induced dose-dependent increases in alpha 6 and beta 1 mRNA levels. TGF beta 1 also increased the expression of laminin beta 2 chain mRNA at 12-16 h poststimulation. In contrast, TGF beta 1 decreased alpha 3 mRNA expression. Immunoprecipitation studies of TGF beta 1-treated cells showed increased surface expression of both alpha 6 and beta 1 protein while surface expression of the alpha 3 integrin subunit was decreased. The same treatment resulted in increased laminin protein expression. These data suggest that TGF beta 1 may regulate alveolar epithelial cell differentiation in part through its modulation of integrins and laminin chains. PMID- 7493639 TI - T cell apoptosis induced by interleukin-2 deprivation or transforming growth factor-beta 2: modulation by the phosphatase inhibitors okadaic acid and calyculin A. AB - The phosphatase inhibitors okadaic acid and calyculin A were used to examine the role of phosphorylation processes in T cell apoptosis induced by interleukin-2 (IL-2) deprivation or transforming growth factor-beta 2 (TGF-beta 2). Okadaic acid and calyculin A inhibited IL-2-driven T cell proliferation and induced apoptosis at concentrations known to inhibit protein phosphatase 1. High concentrations of both agents caused toxic changes of prominent cellular swelling and dilatation of rough endoplasmic reticular profiles. When the T cells were induced to undergo apoptosis by IL-2 deprivation, okadaic acid and calyculin A delayed loss of membrane integrity, nucleosomal size DNA fragmentation, and loss of bcl-2 mRNA. However, T cells deprived of IL-2 in the presence of okadaic acid or calyculin A revealed DNA breaks by in situ DNA end labeling and apoptotic morphology by electron microscopy and failed to show enhanced survival after reexposure to IL-2. Although TGF-beta-mediated signaling is thought to involve the dephosphorylation of specific substrates, okadaic acid and calyculin A not only failed to inhibit, but actually augmented, TGF-beta 2-induced inhibition of T cell proliferation and induction of apoptosis. Exposure to either TGF-beta 2 or the phosphatase inhibitors prevented phosphorylation of the retinoblastoma protein RB. In summary, okadaic acid and calyculin A: (i) induce T cell apoptosis in the presence of IL-2, (ii) allow us to distinguish essential from epiphenomenal features of T cell apoptosis after IL-2 deprivation, and (iii) cooperate with TGF-beta 2 in inducing growth arrest and apoptosis of murine T cells via intracellular cascades that converge in the prevention of RB phosphorylation. PMID- 7493640 TI - Multiple proteases are involved in thymocyte apoptosis. AB - To investigate the involvement of proteases in apoptosis, rat thymocytes were treated with the glucocorticoid hormone methylprednisolone or the topoisomerase II inhibitor etoposide in the presence of selective substrate inhibitors of either interleukin-1 beta-converting enzyme (ICE), (Z-Val-Ala-Asp chloromethylketone, VADcmk) or Ca(2+)-regulated serine protease (Suc-Ala-Ala-Pro Phe-chloromethylketone, AAPFcmk). VADcmk protected from lamin proteolysis, chromatin fragmentation, cell shrinkage, and formation of apoptotic nuclei in both methylprednisolone- and etoposide-treated thymocytes when present during the initiation of the apoptotic process. AAPFcmk prevented lamin breakdown, chromatin fragmentation, and apoptotic morphological changes in thymocytes treated with methylprednisolone, but not with etoposide. Both MPS- and etoposide-treated thymocytes exhibited enhanced ICE-like protease activity which was maximal 1 h after treatment. This increase in proteolytic activity was blocked by VADcmk, but not AAPFcmk. Our findings suggest that ICE-like protease activity is critically involved in the early phase of both methylprednisolone- and etoposide-induced apoptosis in thymocytes, whereas the Ca(2+)-regulated serine protease is an obligatory component of the proteolytic cascade in methylprednisolone-induced apoptosis. PMID- 7493641 TI - Mouse R-cadherin: expression during the organogenesis of pancreas and gastrointestinal tract. AB - Members of the cadherin protein family of cell adhesion molecules are thought to regulate many morphogenetic events during the development of multicellular organisms. We are interested in which cell-cell interactions cadherins regulate during pancreas morphogenesis. Expression and function of E-cadherin in the pancreas have previously been characterized. Recently, R-cadherin cDNA was isolated and the expression patterns of R- and N-cadherin cDNA was isolated and the expression patterns of R- and N-cadherin mRNAs in the pancreas have been described. In the present study, we have investigated the expression of R cadherin in the pancreas and gastrointestinal tract using specific anti-R cadherin antibodies. During early stages of pancreas development (E10.5) R cadherin is expressed in most hormone-producing cells in addition to in primitive duct-like cells. Later, at E18.5, when islets start to form, R-cadherin is not expressed in intraislet endocrine cells, but is instead found in intraductule endocrine and exocrine cells. In adult islets of Langerhans R-cadherin is absent from endocrine cells, but is present in ductule-like cells. Furthermore, the molecule is expressed in exocrine cells: apically and basolaterally on epithelial duct and ductule cells and apically on acinar cells. Interestingly, there appears to be a correlation between the downregulation of R-cadherin expression in intraductule endocrine cells and the migration out of the duct epithelium to form islets. In the gastrointestinal tract R-cadherin is expressed in the pit region of gastric glands and in absorptive cells of the small intestine. The subcellular localization in these tissues is on both the apical and basolateral surfaces. The potential function of R-cadherin, in particular its role in pancreas development, is discussed. PMID- 7493642 TI - Relationship between ornithine decarboxylase and cytoskeletal organization in cultured human keratinocytes: cellular responses to phorbol esters, cytochalasins, and alpha-difluoromethylornithine. AB - Changes in cell shape occur during the cell cycle and influence cell proliferation and differentiation. In order to study how altered cell proliferation and cell shape are interrelated, we have studied ornithine decarboxylase (ODC) regulation in cultured normal human epidermal keratinocytes (NHEK). Cytoskeletal disruptors have been reported to modulate regulation of ODC; the products of ODC, the polyamines, influence actin polymerization rates in vitro, and polyamine auxotrophs have profoundly disrupted cytoskeletons. Therefore, altered ODC levels could be involved in signaling changes in cell shape and an intact cytoskeleton could transduce signals to regulate ODC levels. We had previously observed that the phorbol ester 12-O-tetradecanoylphorbol-13 acetate (TPA), which profoundly alters cell shape, markedly suppresses ODC biosynthesis in NHEK solely at posttranscriptional/protein synthesis levels. TPA treatment caused NHEK to rapidly assume a rounded morphology that was accompanied by a change in actin organization, as determined by rhodamine-phalloidin labeling. Immunolocalization of ODC showed a perinuclear/nuclear distribution in untreated NHEK and a more diffuse pattern after TPA treatment that was apparent within 15-30 min. Changes in ODC enzyme activity are not significant until 60 min after TPA treatment. NHEK treated with cytochalasin B or D to inhibit actin polymerization exhibited a diffuse ODC localization that could be reversed by removal of the cytochalasin; inhibition of ODC by alpha-difluoromethylornithine caused a diffuse ODC localization. All treatments resulted in cytoskeletal remodeling. These data are the first evidence for a distinct subcellular localization for ODC and suggest that changes in ODC localization may be an initial step in regulation of ODC activity. Furthermore, changes in ODC activity cause an altered cytoskeleton, suggesting one means by which growth regulatory signals can be transduced to the cytoskeleton from various signaling pathways. PMID- 7493644 TI - Uptake and metabolism of long-chain 1,2-diacylglycerols by rat adipocytes and H4IIE hepatoma cells. AB - The physiological long-chain diacylglycerols 1,2-dioleoylglycerol and 1,2 dimyristoylglycerol were taken up and metabolized, both by adipocytes and by hepatoma cells, in amounts sufficient to elicit biological responses, without use of emulsifier or any special cell treatment. In adipocytes the only metabolic lipid product of either diacylglycerol was triacylglycerol. In hepatoma cells both diacylglycerols were converted to a number of identified products, mainly phosphatidylcholine and triacylglycerol. This has implications for the interpretation of reports in which long-chain diacylglycerols have been found ineffective in stimulation of cells. PMID- 7493643 TI - Translocation of insulin-regulated glucose transporter is stimulated by long chain 1,2-diacylglycerol in rat adipocytes. AB - The long-chain diacylglycerol 1,2-dimyristoylglycerol emulsified with taurodeoxycholate has been shown to potently stimulate glucose transport in isolated rat adipocytes (Strafors, Nature 335, 554-556 (1988)). We now report that this 1,2-diacylglycerol in the presence of taurodeoxycholate, similarly to insulin, induced translocation of the insulin-regulated glucose transporter (GLUT 4) from a microsomal membrane compartment to the plasma membrane. H4IIE hepatoma cells expressed mRNA for GLUT-1, but not for GLUT-4. In these, otherwise insulin responsive, cells diacylglycerol or insulin had only a marginal effect on glucose transport. PMID- 7493645 TI - Sequestration of PML and Sp100 proteins in an intranuclear viral structure during herpes simplex virus type 1 infection. AB - We investigated the intranuclear distribution of PML and Sp100 in HeLa cells at the ultrastructural level and examined their relocalization in response to herpes simplex virus type 1 (HSV-1) infection. In the absence of infection, we observed that both are components, not only of nuclear bodies, but also of interchromatin granule-associated zones, which suggests a potential role for PML and Sp100 in splicing events. Prolonged HSV-1 infection induced dramatic changes in nuclear organization which consisted of the morphological disappearance of some nuclear structures (nuclear bodies, interchromatin granule-associated zones, coiled bodies) and of the development of a centrally located electron-translucent viral region which pushed the cellular clusters of interchromatin granules to the nuclear border. Concomitantly, dense bodies, concentric arrays of reduplicated inner nuclear membrane, and translucent patches containing a few viral capsids occurred at the nuclear border. PML and Sp100 were exclusively detected over the finely granular material of the viral translucent patches which also contains small amounts of p80-coilin and U1 and U2 snRNAs. An antiserum raised against capsid proteins intensely labeled the viral translucent patches at the level of their finely granular material and enclosed viral capsids. Our data, therefore, suggest that these viral structures, in addition to being the site of accumulation of viral capsid proteins and, possibly, a capsidworks, are also a site of sequestration of cell factors including PML and Sp100. Viral capsid proteins could interfere with and inactivate PML and Sp100 and be implicated in the shutoff of host cell metabolism induced by HSV-1 infection. PMID- 7493646 TI - Different basal NAD levels determine opposite effects of poly(ADP ribosyl)polymerase inhibitors on H2O2-induced apoptosis. AB - We have recently described that poly(ADP-ribosyl)-polymerase (PARP) inhibitors rescue U937 cells from apoptosis induced by 1 mM H2O2 oxidative stress; PARP activation leads to a reversible drop in NAD level, which could be blocked by PARP inhibitors (Nos-seri et al., 1994, Exp. Cell Res. 212, 367-373). A phenotypic variant of U937 is characterized by a lower basal NAD level (low NAD, LN U937, as opposed to the original high NAD, HN U937). In LN cells treatment with 1 mM H2O2, although activating PARP, does not lower NAD concentration; puzzlingly, PARP inhibitors increase (instead of decreasing, as occurs in HN cells) the extent of stress-induced apoptosis, leading to a reduced cell survival. NAD concentration could be increased in LN cells by adding nicotinamide (5-and 25-fold increase) to the culture medium. These cells (LN+) behaved as HN U937: oxidative stress induced a NAD drop, the extent of which is dependent on the cells' basal NAD level; moreover, PARP inhibitors could rescue LN+ cells from peroxide-induced apoptosis. H2O2-induced apoptosis is not triggered by NAD depletion, but instead it takes place only when NAD levels have been preserved or have recovered: on HN U937, peroxide doses (5 and 10 mM) which lead to necrosis induce an irreversible NAD drop, whereas apoptosis occurs only at lower doses, where NAD depletion is reversible; on LN cells NAD levels do not drop even upon 10 mM H2O2 treatment, and these cells die only by apoptosis; moreover, in HN cells apoptosis is not detectable until 8 h posttreatment, when NAD levels recover, whereas in LN cells, where NAD is always present, apoptosis begins to take place as early as 3 h after stress. PMID- 7493647 TI - The increase in H2O2-induced apoptosis by ADP-ribosylation inhibitors is related to cell blebbing. AB - HN and LN are two phenotypic variants of the U937 monocytic cell line which differ in their basal NAD content; they respond in an opposite way to oxidative stress in the presence of the poly(ADP-ribosyl)polymerase (PARP) inhibitors 3 aminobenzamide (3ABA) and nicotinamide (NA): the inhibitors protect HN cells from stress-induced apoptosis, while they enhance it on LN cells (Coppola et al., 1995, Exp. Cell Res. 221, 462-469). These opposite effects are due to two overlapping and contrasting phenomena occurring in LN cells, as shown by the bi modal response of stressed LN cells to increasing 3ABA doses. Indeed H2O2-induced apoptosis is enhanced only at high 3ABA concentrations (i.e., sufficient to inhibit also mono-ADP-ribosylations); lower 3ABA concentrations, which specifically inhibit PARP, also protect LN U937 from stress-induced apoptosis. Unlike HN U937, H2O2-induced apoptosis in LN cells is accompanied by cell blebbing. High 3ABA doses strongly enhance blebbing, leading to cellular fragmentation. Blebbing could be blocked by interfering with actin polymerization with cytochalasin B and D: this eliminated the increase in apoptosis due to 3ABA, suggesting that it is indeed the consequence of excess blebbing. This is supported by the unusual finding that in U937 LN stressed in the presence of 3ABA or NA, blebbing, usually a late event in apoptosis, may even precede its onset. PMID- 7493648 TI - Establishment of a human hepatocyte line derived from primary culture in a collagen gel sandwich culture system. AB - A human hepatocyte line (HHY41) was established from normal human liver tissue. This cell line was derived from a primary culture of human hepatocytes maintained between two layers of collagen gel for 4 weeks. It differs from other human hepatocyte lines in that transfection with the simian virus 40 gene was not used for cellular transformation and nonhepatocellular coculture cells were not present. HHY41 cells have proliferated freely in serum and hormone-supplemented medium after more than 1 year in continuous culture, exhibiting typical morphological characteristics of hepatocytes. HHY41 cells retain glucose-6 phosphatase activity. They also retain the ability to secrete liver-specific proteins such as albumin, transferrin, and alpha-fetoprotein. Northern blot analysis confirmed the presence of albumin mRNA. Cytochromes P450 induced by polycyclic aromatic hydrocarbons are maintained in these cells. Detection of cell surface antigens revealed that HHY41 cells express alpha 1 beta 1-integrin, which is expressed by normal hepatocytes and not by bile duct epithelial cells. High molecular-weight cytokeratin, a marker for bile duct cells, is also absent in HHY41. Cytogenetic analysis showed hyperdiploid karyotype with a consistent deletion in the short arm of chromosome 1. HHY41 can be considered a new human hepatocyte line which retains liver-specific functions of differentiated hepatocytes. Derived from normal liver tissue, not a hepatocellular carcinoma, it provides a new model system for studying the regulation of cell growth and differentiated functions in human hepatocytes. PMID- 7493649 TI - Expression of the human gene encoding urokinase plasminogen activator receptor is activated by disruption of the cytoskeleton. AB - The transition of a stationary cell to a motile cell is associated with the breakdown of the cytoskeleton and expression of enzymes responsible for the degradation of the extracellular matrix. Reverse transcription PCR was used to address the question of whether disruption of the microfilament and microtubule systems is a signal for the increased expression of the urokinase plasminogen activator receptor (uPAR), a key molecule involved in extracellular matrix degradation. Disruption of the cytoskeleton of human fibroblasts with cytochalasin B (1 x 10(-5) M or 1 x 10(-7) M), colchicine (1 x 10(-5) M or 5 x 10(-7) M), or nocodazole (1 x 10(-6) M) resulted in major increases in the levels of uPAR mRNA with smaller increases in uPAR protein levels. Both cycloheximide and alpha-amanitin inhibited the increase in uPAR mRNA levels, demonstrating the requirement for the de novo synthesis of a short-lived protein for transcriptional activation. In contrast to normal fibroblasts, treatment of the metastatic cell line HT1080 with either cytochalasin B or colchicine produced little change in the high levels of uPAR mRNA expressed in these cells. These data suggest that cytoskeletal disruption may contribute to cell migration by signaling an increase in uPAR synthesis. PMID- 7493650 TI - Chondrocyte and chondrosarcoma cell integrins with affinity for collagen type II and their response to mechanical stress. AB - Mechanical stress is an important regulator of chondrocyte functions but the mechanisms by which chondrocytes sense mechanical signals are unknown. Receptors for matrix molecules are likely involved in the mechanical signaling. In the first part of this study we identified integrins with affinity for the cartilage specific collagen type II. We report that the collagen-binding integrins alpha 1 beta 1 and alpha 2 beta 1 isolated from bovine chondrocytes or human chondrosarcoma cells bound collagen type II as judged from affinity chromatography. The integrins alpha 3 beta 1 or alpha 9 beta 1 did not bind collagen type II-Sepharose. In the second part of the study we investigated the effect of mechanical stress on expression of matrix molecules and integrin subunits. Chondrocytes and chondrosarcoma cells, cultured on uncoated flexible silicone membranes in the presence of serum, were exposed to mechanical stress by the Flexercell system. Dynamic stimulation of chondrocytes for 3 h increased the mRNA expression of collagen type II and aggrecan as judged by Northern blotting, while the beta 1-integrin subunit was not changed. When chondrosarcoma cells were exposed to mechanical stimulation under the same conditions, mRNA expression of alpha 5 was found to increase while beta 1, alpha 2, and alpha v did not increase to significant levels. In another study the effect of mechanical stress on integrins was investigated when the cells were cultured on collagen type II coated flex-dishes. Three hours of dynamic stress increased the mRNA expression of alpha 2-integrin subunit while the level of mRNA for integrin subunits beta 1, alpha 1, alpha 5, and alpha v showed no or small changes, indicating that matrix components may modulate the expression of integrins during mechanical stress. PMID- 7493651 TI - Ultrastructural and immunocytochemical characterization of autophagic vacuoles in isolated hepatocytes: effects of vinblastine and asparagine on vacuole distributions. AB - The interactions between the autophagic and the endocytic degradation pathways were investigated by means of immunogold labeling of autophagic vacuoles (AVs) in ultrathin frozen sections from isolated rat hepatocytes. AVs were identified by their autophagocytosed contents of the degradation-resistant cytosolic enzyme CuZn-superoxide dismutase (SOD). Another cytosolic enzyme, carbonic anhydrase (CAIII), was rapidly degraded in the lysosomes, making the vacuolar CAIII/SOD ratio useful as a rough indicator of the progress of autophagic-lysosomal degradation. Lysosomes could be recognized by the presence of the lysosomal membrane glycoprotein lgp120, which was absent from hepatocytic endosomes. Endocytic inputs into the AVs were detected by the presence of gold-conjugated bovine serum albumin (BSA-gold), taken up by fluid-phase endocytosis. All vacuoles recognized morphologically as AVs were SOD-positive, as were essentially all of the lysosomes (96%). The majority (72%) of the lysosomes also labeled positively for BSA within 2 h of endocytosis. The data are thus compatible with the notion that all lysosomes can engage in both autophagic and endocytic degradation. Lgp120 appeared to distinguish well between lysosomes and nonlysosomal AVs: the lgp120-negative AVs (nonlysosomes) had a CAIII/SOD ratio identical to that of the cytosol, indicating that no degradation had occurred. In the lgp120-positive AVs (lysosomes), the ratio was only 43% of the cytosolic value, consistent with substantial CAIII degradation. Among the nonlysosomal AVs (about one-third of all AVs), one-half were BSA-positive, suggesting that early AVs (autophagosomes) and suggesting that early AVs (autophagosomes) and intermediary AVs (amphisomes) that had fused with endosomes were equally abundant. These morphological data thus support previous biochemical evidence for a prelysosomal meeting of the autophagic and endocytic pathways. The microtubule inhibitor vinblastine inhibited the autophagic influx to the lysosomes, causing an accumulation of autophagosomes and a reduction in average lysosomal size. Vinblastine also inhibited the endocytic flux, thereby precluding the formation of amphisomes and of BSA-positive lysosomes. High concentrations (20 mM) of asparagine induced swelling of amphisomes and of BSA-positive lysosomes, probably reflecting an acidotropic effect of ammonia generated by asparagine deamination. Asparagine also caused an accumulation of autophagosomes, amphisomes, and BSA negative lysosomes, presumably as a result of impaired fusion with the swollen BSA-positive lysosomes. The two agents thus appear to perturb the autophagic endocytic-lysosomal vacuole dynamics by different mechanisms, making them useful in the further study of these complex organelle interactions. PMID- 7493652 TI - A new embryonic stem cell line from DBA/1lacJ mice allows genetic modification in a murine model of human inflammation. AB - The development of embryonic stem (ES) cells and their capacity to generate mice with mutations at specific loci has provided a powerful resource for functional analysis of genes in pathological processes. However, the ability to combine this technology with the large number of existing murine models of human genetic disease has been complicated by the inability to routinely generate ES cell lines from strains other than 129. Here, we report the production of a novel ES cell line derived from an inbred mouse, DBA/1lacJ. This new ES cell line undergoes homologous recombination and efficient colonization of the germline of male chimeric offspring with ES cell microinjection into C57B1/6 embryos. The DBA/1lacJ mouse is a murine model of human inflammation, therefore genetic modifications in the DBA ES cells will allow evaluation of the target gene's role in the inflammatory process. PMID- 7493653 TI - Are growth factors chemotactic agents? AB - We report the first direct observation of chemotaxis in slowly moving malignant cells. Two sarcoma cell lines of different metastatic potential were used. In a direct-viewing chemotaxis chamber with two concentric wells, the pooled trajectories of highly malignant rat T15 cells were strongly biased toward the outer well which contained platelet-derived and insulin-like growth factors. In individual experiments, however, the trajectories of the T15 cells showed a significant directional bias which, depending on the cell distribution, sometimes deviated by as much as 170 degrees from the gradient direction. Cells of a less malignant rat line, K2, did not respond to the gradient but a strong K2 response appeared if T15 cells were placed in the outer well along with the growth factors. We conclude that stimulated T15 cells release a chemoattractant, for both T15 and K2 cells, which overrides any chemoattractive effect of the growth factors. These results call into question whether growth factors are ever directly chemotactic in this system and demonstrate the need for direct observation in determining whether any substance is a direct chemoattractant. PMID- 7493654 TI - Dissociation of the stimuli for cell hypertrophy and cell division in the dog thyrocyte: insulin promotes protein accumulation while TSH triggers DNA synthesis. AB - Thyrotropin through the cyclic AMP cascade, and in the presence of insulin or IGF 1, activates the proliferation of dog thyroid cells. In this work it is shown that insulin, presumably acting through the IGF-1 receptors, stimulates the accumulation of protein and increases cell mass but does not induce DNA synthesis. TSH, on the contrary, induces DNA synthesis, in the presence of insulin, but not protein accumulation. It is proposed that insulin induces the increase of cell mass that is a prerequisite for the mitogenic effect of TSH. PMID- 7493655 TI - Cyclin I: a new cyclin encoded by a gene isolated from human brain. AB - A new member of the cyclin family has been isolated from an equalized cDNA library derived from human forebrain cortex. This putative cyclin, designated cyclin I, contains a typical cyclin box near the N-terminus and a PEST sequence near the C-terminus. Cyclin I shows the highest sequence similarity in the cyclin box to cyclins G and E, while the similarity between cyclins I and G also extends toward the C-terminus from the cyclin box. Cyclin I mRNA was expressed at high levels in postmitotic tissues, including skeletal muscle, heart, and brain, and was expressed constantly during cell cycle progression. The expression of cyclin I mRNA does not correlate directly to the cell cycle, and therefore cyclin I may be a novel cyclin member that functions independently of the cell cycle control. PMID- 7493656 TI - Trends of smoking habits in northern Italy (1986-1990). The WHO MONICA Project in Area Brianza, Italy. MONICA Area Brianza Research Group. AB - The trends of age-gender specific prevalence of self-reported smoking habits are presented, observed in two population surveys, performed in 1986-87 and 1989-90 in Area Biranza, a northern Italian industrialized district where a WHO MONICA Centre is located. Methods were internationally standardized to obtain comparable data on two independent random samples, each composed of 1,600 subjects, age-sex stratified and extracted from the 25-64 year old residents. A closed question interview was administered to identify smoking condition (smoker, past-smoker, occasional smoker, never smoker), number of cigarettes consumed per day and attained educational level, categorized in compulsory school and post-compulsory school. Serum thiocyanate was measured as a validation index, using a cutpoint of > 100 mumol l-1 to detect false negatives. Trends in smoking prevalence are analyzed taking into account influences of education in the presence of an anti smoking policy that was started in Italy toward the end of the eighties. Self reported data, confirmed by serum thiocyanate, show a consistent decline of smokers among males (from 48 to 41%), more evident in younger age groups. In females, smoking prevalence is stable (23-24%), although thiocyanate levels in the whole samples indicate a slight but significant tendency to decrease. Education demonstrates positive influences against smoking, particularly in younger male classes. In females a crossover effect is observable: in the second survey youngest group, education results protective against smoking; the contrary is true in the older groups. Our data, detailing what was initially recognized in previous Italian surveys, may be useful to specify the directions of future preventive actions. PMID- 7493657 TI - Twenty-five year mortality from coronary heart disease and its prediction in two Croatian cohorts of middle-aged men. AB - Two cohorts of men aged 40-59 in geographically defined areas of Croatia, former Yugoslavia, were enrolled in 1958 and cardiovascular risk factors were measured. The cohort of Dalmatia, on the Adriatic coast, was made up of 671 men (participation rate 98%), that of Slavonia, in the inland plain, of 696 men (participation rate 91%). Similar examinations were repeated 5 and 10 years after the entry one. A complete follow-up for vital status and causes of death was run for 25 years. Death rates in 25 years from coronary heart disease (CHD) were 90 per 1000 in Dalmatia and 148 per 1000 in Slavonia where also all other major causes of death and all-cause mortality rates were higher (642 vs 465 per 1000 in 25 years). Univariate and multivariate analysis relating ten risk factors to CHD mortality, showed that age and systolic blood pressure were significant predictors in Dalmatia; age, subscapular skinfold and body mass index (inverse) in Slavonia; and age, systolic blood pressure and subscapular skinfold in a multivariate model with lumped cohorts, where dummy variables identifying cohorts indicated a large unexplained extra-risk in Slavonia. In the pooled analysis cigarette smoking and serum cholesterol carried non significant coefficients. Blood pressure change in the first 10 years of follow-up added predictivity to a model exploring the delayed CHD morality occurring between year 10 and 25 of follow-up. Among the base-line measurements only serum cholesterol was significantly different between the two groups (188 mg/dl in Dalmatia and 199 mg/dl in Slavonia).(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7493658 TI - Frequency of consumption of selected indicator foods and serum cholesterol. GISSI EFRIM investigators. Gruppo Italiano per lo Studio della Sopravvivenza nell'Infarto--Epidemiologia dei Fattori di Rischio dell'Infarto Miocardico. AB - The relationship between frequency of consumption of eleven indicator foods (milk, meat, liver, carrots, green vegetables, fruit, eggs, ham, fish, cheese and alcohol) and serum cholesterol was investigated in the comparison group of a case control study of acute myocardial infarction conducted in Italy. Data were collected on 792 subjects from various Italian regions, admitted to hospital for acute conditions unrelated to any known or potential risk factor for myocardial infarction or to long-term modifications in diet. No statistically or epidemiologically meaningful relationship emerged between serum cholesterol level and frequency of consumption of any of these foods. Cholesterol levels rose according to increasing consumption tertiles for most of the indicator foods considered. Higher values for the higher tertile of consumption were observed for meat, ham and eggs, but also for fruit, carrots and green vegetables. However, correlation coefficients between frequency of consumption of various food items and serum cholesterol level were uniformly low for all food items considered, ranging between -0.09 (for milk) and 0.19 (for ham). Although a more comprehensive diet history may lead to different indications, the present data are not suggestive of any major influence of long-term frequency of consumption of a few selected indicator foods on serum cholesterol levels. PMID- 7493660 TI - Prenatal detection of internal urinary system's anomalies. A registry-based study. AB - Prenatal diagnosis performed by ultrasound scan is now a routine part of antenatal care in our region. How many fetal anomalies are actually detected by this procedure? We have used our registry of congenital malformations to answer this question regarding to Internal Urinary System (IUS) anomalies. The mean time of detection of IUS anomalies was 29 weeks of pregnancy (12-39). The sensitivity of prenatal diagnosis of IUS anomalies by ultrasonographic examination was 59.3% for isolated malformations (fetuses with only one anomaly) and 46.0% for multiple malformed children with IUS anomalies. For all cases of IUS anomalies the percentage of prenatal detection was 56.0. Termination of pregnancy was performed in 9.4% of the cases. PMID- 7493659 TI - The influence of dairy products on plasma uric acid in women. AB - Elevated levels of plasma uric acid have been linked to increased risk of cardiovascular diseases and their complications. As dairy proteins have been found to decrease plasma uric acid without increasing glomerular filtration rate, a sample of postmenopausal women living in Montreal was studied to investigate the nature of this relationship. Participants (158 Roman Catholic nuns) were randomly assigned to one of two test diets for a period of four weeks: the dairy foods group (n = 81) consumed approximately 30 grams of dairy protein daily and the dairy-free diet group (n = 77) ate no dairy foods at all. Subjects completed two one-day food records, a core questionnaire and a dairy foods diet history; blood specimens were obtained, and blood pressure, height and weight were measured. Average nutrient intakes differed as a consequence of the test diets, with significantly greater intakes of protein, fat, saturated fat, monounsaturated fat, potassium and calcium (p < 0.01) in the dairy group after the study period, and lower dietary levels of protein, cholesterol, calcium and retinol (p < 0.01) in the dairy-free group. Plasma uric acid was unchanged after the dietary intervention in the dairy group, but increased by 7.8 mumol/l (p = 0.03) in subjects on the dairy-free diet; however, diastolic blood pressure decreased in response to calcium (beta = -22.9, SE = 10.0, p = 0.02) among those whose diet included dairy foods.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7493661 TI - Risk factors of infection and digestive tract colonization by Candida spp. in a neonatal intensive care unit. AB - In this study we determined the risk factors for infection and colonization by Candida spp. in our Neonatal Intensive Care Unit (NICU). We designed a cohort study in the NICU of the La Paz University Hospital. Over a one year period, 153 neonates admitted to the NICU were studied. In the bivariable analysis, hospitalization period, central catheterization, parenteral feeding, parenteral lipid feeding, respiratory support and premature rupture of the membranes (PRM) were statistically associated with infection and colonization; age was only associated with infection. Logistic regression was used to control the confusing factors. The hospitalization period was a risk factor for infection and colonization. PRMs were also colonization risk factors. We developed a statistical equation that predicts the probability of infection or colonization by Candida spp. that are related to a neonate's specific characteristics. The equation helps us to develop preventive procedures. PMID- 7493662 TI - A population survey of childhood accidents in Andalusia (Spain). AB - The aim of this study was to quantify the incidence of childhood accidents and to describe the morbidity profile of accidents in the autonomous region of Andalusia, Spain, for a three-month period (January to March) in 1990. For that purpose, a population survey based on a stratified multiphase sample design was carried out. We collected information about the most serious accident each child had suffered during the study period according to the opinion of the person interviewed (the mother in most of the cases). The cumulative incidence of childhood accidents in Andalusia for a three-month period was 19.4%, higher in urban areas than in rural ones, and lower as the age of the age group increased. As had occurred in previous studies, a higher number of accidents was observed among the male population, however, this result was not statistically significant. During the same period of time, the incidence of traffic accidents for population 0 to 14 years was 0.8%. Accidents among younger children (0 to 4 years of age) occurred mainly at home, while for children of a higher age (5 to 9 and 10 to 14) they took place, in a larger number of cases, either outdoors or at school. When the accident happened, those from the first group were mainly in the presence of their parents or other adults; in the other two groups, there were more friends or schoolmates present. Fractures and injuries, of minor severity, mainly on the head and the extremities and with no sequelae, were the most frequent types of accident.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7493663 TI - The incidence of rubella virus infections in Switzerland after the introduction of the MMR mass vaccination programme. AB - We have collected data on the incidence of rubella in Switzerland from 1987 to 1992 to help evaluating the impact of the measles, mumps and rubella (MMR) mass vaccination programme which started in 1985 in this country. We used detailed informations on samples submitted for diagnostic testing in conjunction with anonymous laboratory notifications to the Swiss Federal Office for Public Health, and data from the Swiss sentinel network of general practitioners to find trends in the incidence of rubella after the introduction of mass vaccination. We observed an unabated seasonal oscillation without decreasing trend during the observation period and were unable to detect a shift in the age distribution of cases. An important proportion of laboratory-confirmed rubella occurred in women of childbearing age. Immigrants from regions with low endemicity of rubella were at increased risk of contracting rubella and transmitting it to their offspring. We conclude, that MMR mass vaccination has not interrupted the circulation of rubella virus in Switzerland, and that improvements in the implementation and surveillance of the MMR vaccination campaign are necessary in order to avoid untoward effects of it. PMID- 7493664 TI - A twelve-year study of leptospirosis on Barbados. AB - Between November 1979 and December 1991, 398 cases of severe leptospirosis were confirmed on Barbados (range for 1980-1991 23-56; mean 32.7; incidence 13.3/100,000/year). For the six-year periods 1980-1985 and 1986-1991 there was no significant change in incidence with time. Incidence is unlikely to change significantly in the next decade. Monthly average case numbers ranged from 1.4 (July) to 4.3 (November). The average (2.8) for June to December (the 7 wetter months) was not significantly higher than that (2.5) for January to May (the 5 drier months). The age range was 7-86. There were three times as many male cases (302) as female (96), and nearly 10 times as many in those < 35. Although the highest number of cases (69) was in males aged 15-24, the highest incidence was in the older age groups, particularly the male 65-74 year-olds, and the female 55 64 year-olds. Leptospirosis was the proven cause of death in 55 (13.8%) hospital patients (annual range 0-13, mean 4.5). Some of a further 39 fatalities might have been cases. Death from leptospirosis was nearly twice as common among the women as among the men. Only one patient under 20 years of age died. Leptospira were isolated and identified from 117 (29.4%) of the 398 sick patients. The infecting organisms were bim (serogroup Autumnalis--75), copenhageni (Icterohaemorrhagiae-26), arborea (Ballum-14) and bajan (Australis-2). These infecting serovars could not be distinguished clinically, but infection was milder in children than in adults. Despite its predominance in surveyed children, serogroup Panama was virtually absent in this study. Rainfall is the major factor affecting the distribution of cases; not surprisingly, sanitation workers and agricultural workers appear to be the groups at highest risk. The general lack of clear-cut risk factors reflects the ubiquity of leptospires in the environment and the fact that the disease is not entirely occupational. PMID- 7493665 TI - Seroepidemiological survey for Borrelia burgdorferi (Lyme disease) in dogs from northwestern of Spain. AB - A random epidemiological study was undertaken to estimate the prevalence of canine borreliosis (Lyme disease) in Castilla y Leon, the largest region in Spain. The presence of antibodies was determined by indirect immunofluorescence assay (IFA), using the Borrelia burgdorferi B31 strain as antigen. Sera from 308 dogs from 7 provinces in the region were tested. Of all the animals sampled, 37 (21%) were seropositive (titres of 1/64 or above). Almost all the provinces had seroprevalence of 20% or below, except for an important focus of borreliosis in the Zamora province (42.8%). Potential risk factors (age, sex, use, habitat, season, and presence of ticks on the animals) relating to the presence of antibodies were evaluated. Those dogs which had at some time had ticks were more often seropositive, at 24.2%, than those which had never had them, at only 6.2% (p < 10-n). No significant differences were discovered for the remaining factors studied. This work indicates that dogs in Castilla y Leon, even if in most cases they do not develop the disease, are exposed to the Lyme disease agent. PMID- 7493666 TI - Molecular marker analysis of Salmonella typhimurium from surface waters, humans, and animals. AB - Salmonella contamination of North Sea water was detected for the first time in 1988 in Germany during routine examinations of bathing areas. Since then, subsequent isolations along the coast have been reported regularly. To define the source of contamination, strains isolated from seawater and rivers were studied by molecular marker methods. Their properties were compared with those of strains originating from possible sources of contamination such as humans, cattle, and sewage treatment plant water. Plasmid profile analysis of whole bacterial populations and the determination of antibiotic resistance patterns demonstrated, that contamination through the surrounding cattle industry could be excluded. Cattle isolates belonged to a widespread clone of phage type 204c which was multiresistant and exhibited an unique plasmid pattern which was never found in sea water isolates. Outer membrane protein and lipopolysaccharide analysis failed to demonstrate differences among the Salmonella populations and proved in this case insufficient for molecular marker discrimination. PMID- 7493669 TI - Geographic spread of Aedes albopictus in Europe and the concern among public health authorities. Report and recommendations of a workshop, held in Rome, December 1994. PMID- 7493668 TI - Epidemiological characteristics of Pseudomonas aeruginosa strains causing infection in an Italian general hospital. A one-year surveillance. AB - During the 1989 calendar year, P. aeruginosa caused clinical infections in 0.46% of patients admitted to Ospedali Riuniti (a general hospital), Bergamo, Italy. Strains (n = 267) of P. aeruginosa were collected during this period, and epidemiological characteristics were studied. The mean prevalence of P. aeruginosa infection in inpatients was 1.1% (range 0.06-7.3), whereas outpatients showed a significantly lower prevalence of infection (0.05%). Strains were recovered from inpatients of surgical wards (n = 126; 47.2%), and outpatients (n = 15; 5.6%). Males were more often affected than females (2.7:1). Infection of the urinary tract was the most common (34.1%). Pseudomonas aeruginosa was also involved in lower respiratory tract infections (18.7%) and septicaemia (17.6%). Four typing methods were performed, i.e. serotyping, antibiotyping, pyocin typing, and restriction endonuclease analysis (REA). Serotypes O:11 and O:6 were endemic in the hospital. Some serotypes correlated with specific clinical wards. Pyocin typing was an unreliable epidemiological tool. However, antibiotyping showed the presence of some epidemic clusters, probably related to the antibiotic consumption of the patients. REA suggested the circulation of edemic P. aeruginosa strains in both the obstetrics and neurosurgery wards. PMID- 7493667 TI - Prevalence of anti-Toxoplasma gondii antibodies in the population of the area of Parma (Italy). AB - Between 1987 and 1991, the prevalence of IgG antibodies to Toxoplasma gondii was determined by ELISA in 28,247 serum samples belonging to 19,432 subjects of the area of Parma (Italy). The overall prevalence was 48.5%. The correlation of infection with age, performed on 420 sera, showed a significant increase of positivity until 30-40 approximately years. In consecutive sera obtained from 172 subjects, the IgG and IgM production was analyzed for about 8 months, and four different patterns were delineated which were comprehensive of the wide range of immunological responses to Toxoplasma gondii exposure observed. Among pregnant females the prevalence of anti-Toxoplasma gondii antibodies was 48.7%, and 5 cases of seroconversion during the pregnancy were observed (0.27%) from which two cases of congenital toxoplasmosis originated. PMID- 7493670 TI - Compliance to compulsory vaccination: strategies and results. AB - This report is devoted to analyze the effect that compulsory vaccination has on the compliance of the population, compared with the results obtained by massive campaigns for optional vaccinations. The implementation of a specific software for the management of individual schedules helps to reach a substantial complete coverage of the individuals for the first ones, while improving but incomplete results regard the vaccines against pertussis and measles-mumps-rubella, optional in Italy. The optimization of data management at the local health unit level improves the quality and the satisfaction of the work performed by the personnel, but has a limited effect on the already nearly complete coverage for the compulsory immunizations. The mounting percentage of children immunized with optional vaccines can be explained both by the massive campaigns of information conducted in recent years and by the better tracking of individual immunization schedules. PMID- 7493671 TI - Coronary heart disease incidence in diabetic patients, and its relationship to general risk factors. AB - The five-year incidence of coronary heart disease (CHD) was examined in middle aged diabetics (54 insulin-dependent--IDDM, 152 non-insulin-dependent--NIDDM) and 105 controls. In comparison to the controls, CHD incidence was higher only in IDDM patients (11.5 vs 4.0%), but the difference was not a significant one. CHD incidence cases in both IDDM and NIDDM groups were significantly related with longer duration of the disease and, when IDDM patients are concerned, with higher diastolic blood pressure. PMID- 7493672 TI - Comparison of parameters to study seasonality in reproductive outcome. AB - This report compares validity, precision and convenience of three frequently used parameters used to describe seasonality in reproductive outcome: prevalence, index and ratio observed versus expected. Each has its advantages and disadvantages; the choice of the parameter depends on the aim of the study. It is advisable to report the number of cases and referents per month, because all three parameters can be derived from this information. PMID- 7493673 TI - Antibodies anti HTLV-I/II in Sicilian residents, in drug addicts, and in African immigrants. PMID- 7493674 TI - Effects of Erwinia-asparaginase on the coagulation system. AB - L-Asparaginase treatment during induction therapy in acute lymphoblastic leukaemia (ALL) is known to be frequently complicated by thromboembolic events. It was recently suggested that L-asparaginase derived from Erwinia chrysanthemi alters the coagulation system less severely than does Escherichia coli asparaginase. In a series of 11 adult patients with ALL, we investigated some parameters of the coagulation system during treatment with Erwinia asparaginase. The doses employed were rather high; all patients below the age of 60 years received 15,000 U/m2 daily over 14 days. In accordance with what is known from treatment with E. coli asparaginase, we observed significant lowering of antithrombin as well as of fibrinogen. However, as to fibrinogen indeed a significant decrease had occurred prior to the institution of Erwinia asparaginase treatment. The most striking observation in the present study was that the levels of prothrombin complex, reflecting the function of K-vitamin dependent coagulation factors II, VII and X, remained within normal ranges during treatment. This indicates that these coagulation factors were not affected by Erwinia asparaginase, an observation at variance with several reports where E. coli asparaginase was investigated. This latter observation was the only finding which could lend support to the view that Erwinia asparaginase affects the coagulation system less than E. coli asparaginase. Finally, one of our patients developed a sinus thrombosis, a severe thrombotic complication. PMID- 7493675 TI - Platelet ultra-structure study in Budd-Chiari syndrome. AB - Morphologic characteristics of circulating platelets were studied in 20 patients with primary Budd-Chiari syndrome (BCS) without any known etiology, using transmission electron microscopy (TEM). Significant platelet ultrastructural changes were observed in all the patients (in 10-->90% platelets) as compared to 20 normal healthy controls (in up to 4% of their platelets). The prominent changes in the platelets were paucity or absence of alpha granules, hypertrophy of the open canalicular systems (OCS) and clumping and fusion of the granules and other organelles in the centre of platelets. Some other changes observed in platelets were dilated channels of OCS, pseudopodial protrusion of cytoplasm and presence of prominent masses of glycogen particles. Platelets from 20 normal controls processed along with the patients' platelets showed only a few such abnormalities. Most of these changes observed in patients' platelets were akin to the changes observed in platelets undergoing activation. Assay of plasma beta thromboglobulin showed significantly higher levels in all the patients (p < 0.001) further confirming on-going in vivo platelet activation with morphologic changes most likely reflecting the thrombotic process present in BCS patients. PMID- 7493676 TI - Retroviral-mediated gene transfer into human bone marrow stromal cells: studies of efficiency and in vivo survival in SCID mice. AB - Retroviral-mediated gene transfer into bone marrow cells is extensively used in gene therapy protocols. Cytokines are needed for stimulation, to achieve a high rate of gene transfer. However, the stromal cell compartment of bone marrow is characterized by rapid proliferation even without cytokines. In this study, human bone marrow stromal cells were isolated and subsequently infected with recombinant retrovirus in a cell-free supernatant. The LN retroviral vector used in this study carries the bacterial neomycin phosphotransferase gene (neoR). Transduction efficiency was significantly enhanced by repeated cycles of infection, with a maximum of 91% transduced cells by four rounds of infection. Presence of the neoR-gene was detected by PCR from all stromal cells selected by G418. After culture in vitro for 3 months, cells were still positive for PCR amplification of the neoR-gene. Transduced stromal cells were also injected into SCID mice to study their homing and survival ability in vivo. PMID- 7493677 TI - Antibodies against platelet GPIb/IX, GPIIb/IIIa, and other platelet antigens in chronic idiopathic thrombocytopenic purpura. AB - Antibodies involved in the pathogenesis of chronic idiopathic thrombocytopenic purpura (ITP) react most frequently with platelet glycoprotein (GP) Ib/IX and GPIIb/IIIa. However, uncertainty as to the specificity, frequency, and clinical significance of such antibodies still remains. By using a modified antigen capture ELISA (MACE), an immunoprecipitation assay, and an immunoblot assay, sera from 60 patients with chronic ITP were analyzed. GP-specific antibodies were found in 50% (30/60) of the patients, with 14 patients having antibodies directed solely to GPIIb/IIIa, 8 holding antibodies specific only for GPIb/IX, and 8 possessing antibodies against both antigens. Serum antibodies were more frequently (p < 0.01) detected in either active and/or non-splenectomized ITP patients. Moreover, in patients displaying antibodies against GPIb/IX, significantly (p < 0.05) lower platelet counts were observed. Using the immunoblot assay, antibodies specific for a 30 kD platelet antigen were detected in 12 of 60 patients. This antigen could not be immunoprecipitated from surface labelled platelet membranes, indicating an intracellular location. We conclude that in chronic ITP, (1) the frequency of anti-GPIIb/IIIa antibodies is close to that of anti-GPIb/IX antibodies, (2) anti-GP antibodies are more likely to be detected in patients with an active disease status and, (3) a 30 kD internal platelet protein is another frequent antigen. PMID- 7493678 TI - Identification of latent myeloproliferative disease in patients with Budd-Chiari syndrome using X-chromosome inactivation patterns and in vitro erythroid colony formation. AB - Some patients with an early or latent myeloproliferative disorder (MPD) present with Budd-Chiari syndrome (BCS, hepatic vein thrombosis). Cell culture analysis of erythroid progenitors (BFU-E) can be used to discriminate primary from secondary MPD and examination of X-chromosome inactivation (in females) can be used to demonstrate clonality in neoplastic tissues. The present study used these techniques to examine whether a group of 7 female patients who presented with BCS had evidence to support a diagnosis of MPD. Unilateral X-inactivation and therefore clonality can be studied in females heterozygous for X-linked restriction fragment length polymorphisms (RFLP) by differences in methylation between active and inactive chromosomes. Probes for two polymorphic loci, phosphoglycerate kinase (PGK, at Xq13.3 [BstX1 RFLP]) and M27 beta (an anonymous locus DXS255 at Xp11.22 [Pst1 RFLP]) were used to study methylation patterns. All 7 patients were heterozygous using M27 beta and 2/7 were also heterozygous using the PGK probe. Polyclonal patterns of X-inactivation in granulocytes were demonstrated in 3/7, a skewed/monoclonal pattern in 1/7 and aberrant patterns in 3/7 using M27 beta. Two patients who had aberrant patterns of X inactivation with M27 beta demonstrated a skewed/monoclonal pattern with PGK. The results of BFU-E growth patterns and clonality were entirely concordant in 5/6 patients. Thus X chromosome inactivation patterns, in conjunction with erythroid colony studies, can be used to assist in the diagnosis of an underlying MPD in BCS. PMID- 7493679 TI - Globin chain turnover in reticulocytes from patients with beta (0) thalassaemia/Hb E disease. AB - beta (0)-Thalassaemia/Hb E disease is the most frequent beta-thalassaemia in Thailand. However, patients have a varying degree of anaemia. The difference in severity is attributed to a differential accumulation of unpaired alpha-globin chains, which is the net result of biosynthesis and catabolism. Turnover of newly synthesized globin chains in reticulocytes from beta(0) -thalassaemia/Hb E patients was determined. Proteolysis was ATP-independent and degraded only 10-15% of the radiolabelled globin during a 4-h incubation period at 37 degrees C and there was no difference in globin turnover in reticulocytes from subjects with mild and severe forms of beta(0)-thalassaemia/Hb E. These results indicate that excess alpha-globin chains do not act as substrates for the reticulocyte proteolytic system and that the difference in severity of anaemia in beta(0) thalassaemia/Hb E disease cannot be explained by a difference in selective post translational catabolism. PMID- 7493680 TI - Increased susceptibility of microcytic red blood cells to in vitro oxidative stress. AB - Oxidative damage to erythrocytes in thalassaemia has been related to generation of free radicals by an excess of denaturated alpha- or beta-globin chains, intracellular iron overload and low concentration of normal haemoglobin (HGB). Two good indicators of such oxidative damage are the high red blood cell (RBC) malonyldialdehyde (MDA) production detected following exogenous oxidant stress and the decrease of pyrimidine 5'-nucleotidase (P5N), the most sensitive enzyme to SH-group damage in vivo. Conflicting data, however, have so far accumulated in the literature concerning differences in oxidative damage between the different forms of thalassaemia and iron deficiency anaemia (IDA). In the present study, oxidative susceptibility, as defined by the production of MDA in vitro and antioxidant capacity, as measured by the activity of RBC glutathione peroxidase (GPx), superoxide dismutase (SOD) and by reduced glutathione (GSH), have been studied in microcytic RBCs from patients with beta-thalassaemia trait, Spanish (delta beta) zero-thalassaemia heterozygotes (delta beta-thalassaemia trait) and iron deficiency anaemia (IDA). The results are consistent with the existence of significant differences in the severity and pattern of oxidative stress susceptibility between beta-thalassaemia trait (increased MDA production and higher SOD and GPx activities) and the other two forms of microcytosis (delta beta thalassaemia trait and IDA). Furthermore, the finding of normal P5' N activity in delta beta thalassaemia trait, gives further support to the less intense peroxidative environment of RBCs in this form of thalassaemia when compared to beta-thalassaemia trait, characterized by acquired RBC P5' N deficiency due to oxidative damage. PMID- 7493681 TI - Multiple myeloma associated with Epstein-Barr virus in an AIDS patient: a case report. PMID- 7493682 TI - Spontaneous adrenal gland haematoma in a patient with antiphospholipid antibodies. PMID- 7493683 TI - Interleukin-1 beta and interleukin-6 in clonal versus reactive thrombocytosis. PMID- 7493684 TI - Association of a myelodysplastic syndrome with hairy cell leukaemia. PMID- 7493685 TI - Clinical usefulness of bronchoalveolar lavage in haematologic patients with suspected pulmonary infection. PMID- 7493686 TI - Effects of granulocyte-macrophage colony stimulating factor produced in Chinese hamster ovary cells (regramostim), Escherichia coli (molgramostim) and yeast (sargramostim) on priming peripheral blood progenitor cells for use with autologous bone marrow after high-dose chemotherapy. AB - Peripheral blood progenitor cells (PBPCs) were collected without prior association with chemotherapy but after the administration of granulocyte macrophage colony-stimulating factor (GM-CSF) produced in Chinese hamster ovary cells (CHO-GM, regramostim), Escherichia coli (E. coli-GM, molgramostim), or yeast (Yeast-GM, sargramostim) and used in conjunction with autologous bone marrow after high-dose chemotherapy in 69 patients with breast cancer or melanoma. The mean peripheral white blood cell (WBC) counts increased by 2.2 to 2.7-fold after regramostim, 4.5 to 7.3-fold after molgramostim and 4.3-fold after sargramostim. All patients underwent three leukaphereses. The mean (+/- standard error) total nucleated pheresed cells per kg x 10(8) were 4.15 +/- 0.56, 15.10 +/ 1.77 and 7.24 +/- 1.00 for patients receiving regramostim, molgramostim or sargramostim respectively. The mean (+/- standard error) granulocyte-macrophage colony-forming units per kg x 10(4) mobilized into the PB were 8.75 +/- 3.63, 71.03 +/- 17.85, and 65.11 +/- 18.74 for patients receiving regramostim, molgramostim, or sargramostim respectively. The total mean (+/- standard error) CD34+ cells per kg x 10(7) collected by three leukaphereses were 3.28 +/- 1.62, 1.34 +/- 0.51 and 2.57 +/- 1.93, for patients receiving regramostim, molgramostim or sargramostim respectively. The use of either molgramostim- or sargramostim primed PBPCs led to complete elimination of absolute leukopenia with a WBC count under 100/mm3 in 64% and 77% of patients treated, respectively. Patients receiving molgramostim-primed PBPCs required fewer red blood cells transfusions than patients receiving regramostim-primed PBPCs (p = 0.0062). Our data indicate that PBPCs collected without prior association with chemotherapy but after either molgramostim or sargramostim with autologous bone marrow support and GM-CSF shorten the hematopoietic recovery after myeloablative chemotherapy in patients with breast cancer or melanoma. PMID- 7493687 TI - Thriving on change: strategies for success. PMID- 7493688 TI - Conflicting Doppler data on the assessment of pelvic tumors vascularity. PMID- 7493689 TI - Review of the endometrial safety during intravaginal treatment with estriol. AB - To gain more insight into whether intravaginal treatment of local urogenital complaints with the mild-acting oestrogen estriol is capable of inducing proliferation of the endometrium, the results of the clinical studies that have been published over the years have been pooled. Of a total of 19 studies that initially had been selected, four were excluded from the analysis because no baseline biopsies were available, two because endometriae had been evaluated using methods other than with histology, and one study because a sustained release preparation was used. Pooling of 12 studies (214 subjects) revealed a reasonable amount of long-term data on intravaginal estriol treatment with 61 evaluable biopsies after 6 months and 58 after 12 months. In addition, 13 biopsies were available after 2 years. It appeared that intravaginal estriol treatment using the recommended dosages did not result in endometrial proliferation. All 337 post-baseline biopsies that have been reported in the literature were classified as atrophic. It can be concluded that single daily treatment with intravaginal estriol in the recommended doses in postmenopausal women is safe and without an increased risk of endometrial proliferation or hyperplasia. Consequently, there is no need to add sequential progestogens with these preparations and no withdrawal bleedings will be induced. PMID- 7493690 TI - The transport of vitamin C in the isolated human near-term placenta. AB - We investigated the specific uptake (reference: [H3]-L-glucose) as a measure of membrane transfer of [C14]-labeled L-ascorbic acid (AA), L-dehydroascorbic acid (DHA) and diketogulonic acid using the single injection, double tracer dilution method in the artificially perfused lobe of the near-term human placenta. The uptake of DHA (40-60%) on both the fetal and the maternal side was 3-6 times higher than the uptake of AA, whereas an uptake of diketogulonic acid was not detected. AA transport was slightly higher on the maternal side. Uptake of DHA was suppressed by phloretin and cytocholasin B, whereas AA transfer was not affected. Low sodium concentrations inhibited the uptake of DHA. D-glucose (> 20 mmol/l) inhibited the DHA uptake, and DHA inhibited D-glucose uptake but not L alanine uptake. The Km value (self inhibition) for DHA was 6-14 mmol/l. Vitamin C enters the trophoblast predominantly as L-dehydroascorbic acid. Its transfer through the microvillous and basal membrane might use the glucose transporter, however, a specific sodium-dependent pathway is not ruled out. Our transfer data suggest an intracellular pool of vitamin C which fills up with increasing plasma DHA-concentrations. PMID- 7493691 TI - Energy economy of human uterine muscle strips under different in vitro conditions and its dependence on tissue redox potential. AB - OBJECTIVE: To study the energy economy of human myometrium in vitro. STUDY DESIGN: Myometric biopsies were kept for 10 min or 4 h in a buffer solution at +4 degrees C before preparation into 'early' and 'late' strips, respectively. The strips were perifused at +37 degrees C for different periods of time. Adenylates, energy charge (EC), adenosine and NAD+ were determined in individual strips. RESULTS: Mean EC and ATP values were low, the 'late' strips displaying 5% and 20% lower values than the 'early' strips, respectively. During perifusion the EC increased slightly, ADP decreased significantly and the strips began to contract. A correlation fitting a multiplicative function was evident between EC and NAD+ (r = 0.89, P < 0.001). CONCLUSION: Human myometrium, being operative at an unusually low EC level, maintained its energy economizing and contractile functions in vitro for 4 h, displaying a tight relationship between EC and NAD+, suggestive of a compromised energy balance. PMID- 7493692 TI - Adnexal torsion: the contribution of color Doppler sonography to diagnosis and post-operative follow-up. AB - Ovarian torsion is often difficult to diagnose due to non-specific clinical and clinical and ultrasonographic findings. We demonstrated the use of color Doppler for the early diagnosis of ovarian torsion and for post-operative follow-up until the normalization of the preserved ovary was confirmed by subsequent viable pregnancy. Color Doppler flow assessment can be useful as both a diagnostic tool and for evaluating the recovery of the affected ovary after conservative surgical treatment. PMID- 7493693 TI - Pyogenic sacro-iliitis, a rare cause of peripartum pelvic pain. AB - A patient was presented suffering from severe pelvic pain several weeks after delivery. Symptoms pointed in the direction of a peripartum pelvic pain syndrome. One week after admission, the clinical course deteriorated. An infective endocarditis complicated by pyogenic sacro-iliitis was diagnosed. PMID- 7493694 TI - Acute HELLP postpartum with renal failure. PMID- 7493695 TI - Combined simple and complex cord knots. AB - Both double true knots and complex knots of the umbilical cord are rare events. This report presents a case in which the two occurrences were combined. Apart from some early decelerations in the first stage of labour, there was no evidence of fetal compromise. A mid-cavity forceps delivery performed for poor maternal effort resulted in the birth of a healthy female infant. The mechanism and consequences of umbilical cord knotting are reviewed. PMID- 7493696 TI - Two cases of placenta accreta managed conservatively. AB - Placenta accreta is a rare complication of pregnancy. Traditionally treatment has been operative, commonly total abdominal hysterectomy, in order to prevent serious haemorrhage or infection. Reproductive function can, however, be preserved by conservative management which is possible in carefully selected cases without risking maternal welfare. We report two cases of placenta accreta managed conservatively--both women delivered after this treatment. PMID- 7493697 TI - Late complication of ureteric damage following hysterectomy. PMID- 7493698 TI - Wilson's disease in pregnancy. AB - Patients with Wilson's disease contemplating pregnancy should have their hepatic function and copper status assessed. We report a case of a pregnant woman with Wilson's disease with compromised hepatic function. The medical problems and controversy of prescribing treatment are discussed. PMID- 7493700 TI - Postgraduate training in obstetrics and gynaecology. PMID- 7493699 TI - Biochemical and Doppler predictors of poor perinatal outcome in a fetus with four umbilical vessels. AB - Unexpected elevation of maternal serum alpha-fetoprotein (MSAFP) in women with anti-phospholipid syndrome has already been described as a predictor of fetal death. In this report we present a case of a pregnant woman with elevated second trimester MSAFP, in which Doppler ultrasound at 28 weeks suggested very poor fetal prognosis. A cesarean section was performed, but 2 days later the infant died due to distress. The only remarkable feature at post-mortem study was the finding of four vessels in the umbilical cord. Conventional investigation of the mother led us to the diagnosis of a primary anti-phospholipid syndrome. The finding of such an association should alert clinicians to the increased risk of fetal death. Precocious Doppler ultrasound examination may be the elective non invasive technique to monitor such high risk fetuses. PMID- 7493701 TI - Shoulder dystocia--is it predictable? AB - An unmatched comparative study is described to determine if routine clinical indicators are useful predictors for shoulder dystocia. Parity, maternal weight gain during pregnancy, and a history of a previous large baby and increased operative vaginal delivery rate were more often associated with shoulder dystocia. No other significant associations were found. However, shoulder dystocia can not be predicted accurately antepartum using routinely available clinical factors. PMID- 7493702 TI - Breech presentation in Spain, 1992: a collaborative study. AB - OBJECTIVES: The goals of the present work were to study the incidence of breech deliveries in Spain, the causes for breech presentation and its management and the incidence of cesarean sections and its correlation with the perinatal mortality rates among different centers. STUDY DESIGN: Data from 102,038 deliveries, 3829 of which were singleton breech deliveries attended in 1992, were obtained from 40 hospital centers located in different autonomic communities of Spain and analyzed using a computer statistical package (R-sigma, Horus Hardware). RESULTS: The incidence of breech deliveries was 3.8%, although this proportion varied between different centers and regions. Up to 18% of the cases occurred in preterm. The most frequent causes were prematurity and uterine malformations. In its management, an increase in the tendency to perform elective cesarean section was noticed. The incidence of cesarean sections reached 14.9% of the total and 67.9% in breech deliveries. Perinatal mortality rate in breech deliveries was more than four times that of all deliveries, according to national standards, and more than three times according to international standards (47.8 versus 11.8 x 1000, and 24.7 versus 8 x 1000, respectively). The intrapartum and neonatal mortality in breech deliveries was 13 x 1000. No correlation between incidence of cesarean sections and perinatal mortality rates was found. Primiparity, deflexed head, macrosomia and prematurity were the most decisive parameters to indicate a cesarean section. Neither the external cephalic version nor the total breech extraction were common procedures. CONCLUSIONS: The incidence of breech deliveries is variable between different centers (1.7%-5.1%) and Spanish regions (3.3%-4.3%), with a mean of 3.8% in overall and 11.4% in preterm deliveries. There is a tendency to perform cesarean sections, especially in primiparae. There is no correlation between the percentage of cesarean sections in each center and the perinatal mortality rate. External cephalic version and total breech extraction were uncommon procedures. PMID- 7493704 TI - Cesarean section rates and perinatal outcomes in resident and midwife attended low risk deliveries. AB - Cesarean section rates and perinatal outcomes of low risk pregnancies were evaluated in two different Turkish state hospitals. In this retrospective study, we compared cesarean section rates of 1668 low risk and non-private pregnancies managed by residents (group 1) and midwives (group 2), 53.3% and 46.6%, respectively. All patients belonged to a low socioeconomic class and none of them paid hospital costs themselves. Patient characteristics, mode of delivery and perinatal outcome information were studied by blind reviewers. Although patient characteristics and perinatal outcomes among the two groups were not statistically significant (P > 0.05), the cesarean section rate was three times higher in the resident group (9.88% vs. 3.08%) and this difference was statistically significant (P < 0.05). These findings suggest that reducing the expanded use of cesarean section for low risk pregnancies in teaching hospitals would not have an adverse effect on perinatal outcome. PMID- 7493703 TI - Intra-uterine infection in women with preterm premature rupture of membranes: maternal and neonatal characteristics. AB - OBJECTIVE: Our purpose was to determine the prevalence of intra-uterine infection in patients with preterm premature rupture of membranes and to evaluate the clinical characteristics of women and neonates according to the presence or absence of intrauterine infection. STUDY DESIGN: Trans-abdominal amniocentesis was performed in 90 consecutive patients admitted with preterm premature rupture of membranes. Maternal clinical parameters evaluated included maternal age, origin, gravidity, parity, habitual abortion, previous perinatal death, previous preterm birth, urinary tract infection, fetal distress, abruptio placentae, gestational diabetes, admission-delivery interval and cesarean rate. Neonatal clinical parameters evaluated were gestational age at delivery, birth weight, Apgar score, neonatal gender, perinatal death and neonatal sepsis. RESULTS: The prevalence of intra-uterine infection was 66.6% (60/90). No differences between patients with intra-uterine infection and those without intra-uterine infection were observed in maternal age, origin, gravidity, parity, habitual abortion, previous perinatal death, fetal distress, abruptio placentae, gestational diabetes, admission-delivery interval, cesarean section, neonate's gender, Apgar score at 5 min and neonatal sepsis. On the other hand, gestational age at delivery, birth weight and Apgar score at 1 min were significantly different between the groups. CONCLUSION: An intra-uterine infection was found in two thirds of women presenting with preterm premature rupture of membranes. Women presenting with preterm premature rupture of membranes and intra-uterine infection had a lower gestational age at delivery and consequently had a lower neonatal birth weight than those without intra-uterine infection. PMID- 7493705 TI - Colposcopy and directed biopsy reliability during pregnancy: a cohort study. AB - The reliability of cytology, colposcopy, and directed biopsy were assessed in pregnancy and compared with that observed in a matched non-pregnant control group in order to evaluate the influence of pregnancy. One hundred and seventeen gravidas in which the final pathology could be ascertained and 234 control patients were examined by the same colposcopist. Concordance between cytology and final diagnosis was complete or within one degree of severity in 55.6% and 77.8% of patients, respectively. Unsatisfactory colposcopy was significantly less frequent in pregnant patients than in controls (12.8% vs. 23.1%, P = 0.023). In pregnant patients, colposcopy provided concordance, overestimation and underestimation of the final diagnosis in 72.6%, 17.6% and 9.8%, respectively. In the same group, the concordance between the histologic findings of directed biopsy and final diagnosis was complete or within one degree of severity in 83.7% and 95.9%, respectively. The reliability of cytology, colposcopy and directed biopsy was not related to pregnancy. These data show that the physiological changes which occur in pregnancy do not significantly alter the reliability of colposcopy and directed biopsy, if the colposcopist is aware of the peculiar difficulties and does not overreact to the accentuated patterns that may occur during pregnancy. PMID- 7493706 TI - Trans-vaginal ultrasound examination of ovarian masses in premenopausal women. AB - OBJECTIVE: Two scoring systems based on trans-vaginal sonographic findings of ovarian tumors were evaluated with respect to the prediction of ovarian malignancy. STUDY DESIGN: In this retrospective study, 64 premenopausal non pregnant patients with an ovarian tumor underwent trans-vaginal sonography from 1991 to 1993. In each women a numerical score was calculated using two methods. Method A included four variables: inner wall structure, wall thickness, presence of septa and echogenicity. The point scale ranged from 4 to 15 points. Method B included three morphological characteristics: volume, wall structure and septal structure, the point scale ranging from 0 to 12 points. The threshold values for predictability of malignancy were 9 and 5 points for methods A and B, respectively. RESULTS: The mean age of the patients was 32.2 years (range 22-44). The mean gravidity was 2.6 (range 0-5) in 45 women, 19 women being nulliparous. Fifty-five masses were surgically proven to be benign and 9 women had primary malignant tumors. Sensitivity and specificity for prediction of malignancy were 89% and 73%, respectively, for method A, and 89% and 70%, respectively, for method B. Positive predictive value and negative predictive value for malignancy were 35% and 97% for method A and 29% and 64% for method B. The mean morphology scores on benign and malignant masses, calculated by method A, were 6.94 +/- 2.36 S.D. and 12.0 +/- 2.4 S.D. (P < 0.01). The same scores, calculated by method B, were 4.16 +/- 1.16 S.D. and 9.44 +/- 2.96 S.D. (P < 0.01). We found a considerable overlap in the scores of different types of ovarian tumors. CONCLUSIONS: Both methods are easy to apply and provided explicit data. Method A was shown to be more effective. The number of false positive results was relatively high. The most important single sonographic characteristic of the malignant masses are the wall structure abnormalities. PMID- 7493707 TI - Abdomino pelvic irradiation after second-look laparotomy for stage III ovarian carcinoma. AB - OBJECTIVE: The purpose of this retrospective analysis of 34 patients with stage III ovarian carcinoma was to review results and morbidity of whole abdominal irradiation after surgery and chemotherapy. METHODS AND MATERIALS: All of the 34 patients had reached a complete clinical remission after first cytoreductive surgery and chemotherapy. After second-look laparotomy each patient underwent whole abdominal irradiation. Except for two patients with chronic myelosuppression, the dose administered was of 22.5 Gy to the abdominal cavity with a boost of 22.5 Gy added to the pelvis. RESULTS: Three and 5-year overall survival rates were 62% and 43%, respectively. Three and 5-year disease-free survival rates were 53% and 38%. Twenty-three patients (68%) developed local relapse or local disease progression. Metastasis occurred in five cases and were always associated with an abdominal cavity recurrence. Residual disease after first cytoreductive surgery appeared as a prognostic factor in univariate analysis. Patients with unresected residuum had a 5-year survival probability of 35% versus 83% for patients without residual disease. We observed 12% grade-3 intestinal toxicities and one fatal case of radiation enteritis. CONCLUSION: Despite its curative potential, the long term benefit of whole abdominal irradiation in the multimodality treatment of advanced ovarian carcinoma must be evaluated in well designed controlled trials. PMID- 7493709 TI - Cervical polyp in the menopause and the need for fractional dilatation and curettage. AB - OBJECTIVE: To investigate the need for fractional dilatation and curettage following excision of symptomatic versus asymptomatic cervical polyps. STUDY DESIGN: A prospective study was carried out on 467 women who were referred for treatment of symptomatic (accompanied by vaginal bleeding or discharge) or asymptomatic cervical polyps, from January 1, 1990 to December 31, 1992. Of these, 204 were premenopausal and 263 postmenopausal. Every excision of a cervical polyp was followed by a dilatation and curettage. The histological data were evaluated statistically using the chi 2-test. RESULTS: Postmenopausal women had more asymptomatic than symptomatic cervical polyps (P = 0.004). Cervical polyps were associated with more endometrial polyps in the postmenopausal than in the premenopausal women (P = 0.0009). Postmenopausal women with symptomatic cervical polyps had more endometrial abnormalities on histological examination than those with asymptomatic ones (P < 0.0001); this difference was not significant in the premenopausal group (P = 0.49). CONCLUSIONS: While neither symptomatic nor asymptomatic cervical polyps are an indication for dilatation and curettage (following excision) in women in their reproductive years, and do not affect their management or prognosis, this is not the case in postmenopausal women. Symptomatic cervical polyps after the menopause must be excised and followed by mandatory fractional dilatation and curettage, because there is a marked incidence of associated severe pathological conditions in this age group. PMID- 7493708 TI - Management of low-grade CIN: follow-up or treatment? AB - OBJECTIVE: To examine whether low-grade cervical dysplasia carries a higher risk of progression when associated with the cancer-related human papillomavirus types 16, 18, 31 or 33. STUDY DESIGN: Retrospective, with PCR-based HPV diagnosis on the original cervical biopsies from 71 patients with CIN I and II. CIN III developed in 34 lesions, and 37 showed complete regression during non-invasive follow-up. RESULTS: Progression occurred in 15/41 CIN I and in 19/30 CIN II lesions (P = 0.03). HPV DNA was detected in 43 specimens. CIN III developed in 25% of HPV-negative lesions, in 48% of HPV-positive CIN I lesions, and in 77% of HPV-positive CIN II lesions. CONCLUSION: Low-grade lesions are at higher risk of progression when associated with HPV types 16, 18, 31 or 33 (P = 0.002). HPV diagnosis can be useful in the triage of patients with low-grade CIN. PMID- 7493710 TI - Polycystic ovaries in association with mullerian anomalies. AB - The polycystic ovary syndrome, whose etiopathogenesis is not clearly understood, has a wide spectrum of clinical presentations, and may co-exist with other pathologic conditions. In this study, we evaluated the prevalence of ultrasound defined polycystic ovaries (PCO) in patients with mullerian anomalies (n = 167), and those without mullerian anomalies (n = 3165) from 1990 to 1994, in a population markedly composed of infertility patients. PCO were found in 50 (29.9%) patients in the study group, compared to 637 (20.1%) patients in controls (P < 0.01). Mullerian anomalies were further grouped according to the American Fertility Society (AFS) classification and it was found that patients with the septate uteri and bicornuate uteri malformations had a higher prevalence of PCO than the controls (P < 0.001, P < 0.05, respectively). Although a difference existed in the percentage of PCO in patients with unicornuate uteri and didelphic uteri compared to controls, this difference did not reach statistical significance (P > 0.05). We conclude that, as PCO are more prevalent in certain mullerian anomalies, an embryogenetic defect may also be involved in the etiopathogenesis of PCO. PMID- 7493711 TI - Ectopic pregnancy and laparoscopy: review of 1197 patients treated by salpingectomy or salpingotomy. AB - The past few decades have witnessed such a rapid rise in the incidence of ectopic pregnancy that it verges on the point of an 'epidemic disease'. Its early detection, with the aid of serum beta-hCG, high resolution ultrasound and the more liberal use of laparoscopy, has dramatically altered the clinical presentation of this disease and permits the use of more conservative methods of management directed towards preserving fertility and reducing morbidity. In this review of 1197 patients, compiled from the English literature, various conservative or tubectomy operative laparoscopic procedures have been employed, with 93% and 98% respectively, being able to avoid further surgery. Among the group treated by the conservative approach, a 6% post-operative complication rate was reported, of which 4% were persistent ectopic, 48% intra-uterine, and 18% repeated ectopic pregnancies. Among the radically treated patients, 2 intra- and 1 post-operative complications necessitated laparotomies. The fertility work-up and performance outcome are less obvious among this group. The benefits, safety and efficacy of each of the laparoscopic options, with appropriate recommendations for their use, are discussed. However, despite the aforementioned dramatic progress, women with previous ectopic pregnancies still have reduced fertility potential. Preventive measures aimed at reducing its overall occurrence therefore seem to be the major factor towards preserving a patient's future fertility potential. PMID- 7493712 TI - No evidence of the hook effect in peritoneal fluid CA 125 measurement using immunoenzymatic second generation assays: comparison with immunoradiometric assays. AB - OBJECTIVE: To monitor the occurrence of the hook effect in measurements of peritoneal fluid CA 125 levels using two different immunoenzymatic second generation assays (ETI-II and EIA-II), and to compare these results with those obtained using the respective immunoradiometric versions of the assays (IRMA-II and ELSA-II). STUDY DESIGN: CA 125 levels were determined in peritoneal fluid and serum samples obtained from 45 women with gynecological diseases. The assays were carried out using IRMA-II and ETI-II (Sorin Biomedica) and ELSA-II and EIA-II (CIS Bio International) assays. Occurrence of the hook effect and linearity of the assays were evaluated. Statistical analyses were performed by Wilcoxon's test and linear regression analysis. RESULTS: Undiluted peritoneal fluids, assayed for their CA 125 content, showed falsely underestimated values of the antigen when IRMA-II and ELSA-II assays were performed. The phenomenon disappeared only at high dilutions of the sample (> 50). Conversely, immunoenzymatic ETI-II and EIA II assays performed on undiluted peritoneal fluids did not show underestimated CA 125 values. CA 125 values obtained by immunoenzymatic assay were lower than those obtained using their respective immunoradiometric versions at a dilution of 1:100 (P < 0.001). A good correlation was observed between ELSA-II and EIA-II (r = 0.929) CA 125 values. CONCLUSION: The EIA-II immunoenzymatic assay appears to be more suitable for CA 125 measurement in peritoneal fluid in that it is not subject to the hook effect and its results correlated well with those obtained via its immunoradiometric version. PMID- 7493713 TI - The management of pregnancy and delivery in HIV-infected women in Europe. AB - In view of the increasing heterosexual spread of HIV and reports of interventions which reduce the rate of mother to child transmission, a postal questionnaire survey was performed to determine the strategies for the management of pregnancy and delivery in HIV-infected women in 56 obstetric centres in 21 European countries. A response rate of 96% was achieved. Antenatal testing for HIV infection was routinely offered in 96% of centres. Pre- and post-test counselling was available in most centres and was mainly provided by the obstetrician; only 12 centres had trained counsellors. Compared to uninfected women, infected women made more antenatal visits in 36% of centres and had more screening tests in 30% of centres. Some delivery procedures (e.g. use of scalp electrodes) were less common in infected than in uninfected women. Nearly half the centres prescribe Zidovudine to infected women; most prescribed it specifically to reduce HIV vertical transmission. Disinfection of the birth canal and elective caesarean sections for infected women were routinely performed in 22% and 18% of centres, respectively. The survey revealed different approaches to the management of infected pregnant women both between and within countries, which may partly be due to limited knowledge of what constitutes optimal management, especially in relation to interventions to reduce vertical transmission. Randomised control trials are required to establish the most effective intervention. PMID- 7493714 TI - Hysteroscopic endomyometrial resection for the treatment of menorrhagia--follow up of 86 cases. AB - OBJECTIVE: The aim of the study was to determine the effectiveness of hysteroscopic endomyometrial resection in the treatment of menorrhagia. STUDY DESIGN: Eighty-six women with menorrhagia underwent hysteroscopic endomyometrial resection between November 1990 and November 1993. The surgery was carried out under spinal anesthesia in 54 (64%) of the patients. A standard resectoscope was used and the uterine cavity distended with 2.2% glycine. The mean follow-up period was 22 months (range 4-39). RESULTS: No major complications occurred and no fluid overload was reported. One episode of uterine perforation, one of pelvic infection, five episodes of postspinal headache and four of hematometra were recorded. Hysterectomy was performed in five cases. Follow-up information was available on 79 (92%) patients. Amenorrhea was achieved in 20 (25%) patients and hypomenorrhea in 44 (56%) patients. Only three (4%) patients showed no improvement in menstrual flow. CONCLUSIONS: Hysteroscopic endomyometrial resection is a safe and effective mode of treatment for menorrhagia in selected patients. PMID- 7493715 TI - Experience and opinions of climacterium by Finnish women. AB - OBJECTIVE: Literature on women's experiences with climacterium is mostly based on patients or on North American women. In this study, the experiences and opinions of Finnish women were investigated. STUDY DESIGN: In 1989, a questionnaire was sent to a representative sample of 45-64-year-old Finnish women (n = 2000); 1713 (86%) responded, of which 1308 were postmenopausal. RESULTS: Most women (87%) reported their health to be good or rather good. Symptoms were commonly experienced, but each symptom usually by a minority of women. Most subjective health problems were not related to climacterium as such, but were problems also encountered otherwise or related to aging. Some symptoms decreased with increased age, others increased or stayed the same. Comparing the symptoms reported in the past two weeks and women's own judgements suggests that hot flashes and irritability were specific to climacterium. Most women had a neutral or positive opinion of climacterium in general. CONCLUSION: Our study challenges the view that climacterium is a time of big subjective health problems. PMID- 7493716 TI - Effect of a new cyclical sequential postmenopausal HRT on lipoprotein, apoprotein and thrombophilia profile. AB - We examined the changes in lipoprotein, apoprotein, and thrombophilia profile in postmenopausal women using a new cyclical sequential combined HRT regimen. The study medication consisted of two tablets of Hormonin (oestriol 0.27 mg, oestrone 1.4 mg and oestradiol 0.6 mg), daily and norethisterone (1 mg) BP (Shire Developments) for the last 12 days of every 28 day cycle. Serial fasting blood samples were collected at the beginning of the study and, thereafter, at 3 monthly intervals for 1 year, each patient acting as her own control. Thirty-five healthy postmenopausal women completed 1 year of follow-up and had a complete set of fasting blood samples. The lipid profile; total cholesterol, triglycerides, HDL, LDL, Apo AI, Apo AII, Apo B and Lp(a), as well as the coagulation parameters; antithrombin III, factor VII, fibrinogen, protein C and protein S, were measured at each occasion. There was a statistically significant drop in total cholesterol and LDL levels. Lp(a) level dropped after commencing treatment and remained below baseline for the rest of the study. The initial increase in Apo AII was not maintained for the duration of the treatment. The changes in Lp(a) and Apo AII were not statistically significant. The level of protein S dropped significantly throughout the study. The changes in other coagulation factors were not statistically significant. The effect of this hormonal combination on the lipid parameters is favourable, and although the change in protein S is striking, its clinical significance remains unclear. PMID- 7493718 TI - Time, costs and meeting needs. PMID- 7493717 TI - Correlations between periovulatory serum and follicular fluid CA-125 and granulosa cell hormones after controlled ovarian hyperstimulation. AB - OBJECTIVE: To determine the presence and concentration of CA-125 in periovulatory follicular fluid (FF) and serum after controlled ovarian hyperstimulation and to determine if the CA-125 in these two compartments could be related to granulosa cell markers such as inhibin or estradiol. STUDY DESIGN: Fifteen women undergoing controlled ovarian hyperstimulation for in-vitro fertilization-embryo transfer were studied. A transvaginal, ultrasound-guided follicular aspiration was performed. CA-125, inhibin, estradiol and FSH were measured in FF and serum. Pearson and Spearman's Rank Correlation tests were performed. RESULTS: CA-125 was measurable in 59% of follicles. Values ranged from undetectable to 3630 U/ml. Serum CA-125 ranged from undetectable to 126 U/ml. CA-125 and inhibin correlated negatively in FF and positively in serum. CONCLUSION: CA-125 was present in significant but variable concentrations in 59% of periovulatory follicles. A negative correlation was noted between CA-125 and inhibin or estradiol in the FF and a positive correlation with serum inhibin. No correlations were noted to oocyte retrieval or fertilization. PMID- 7493719 TI - SPECT of the brain in childhood autism: evidence for a lack of normal hemispheric asymmetry. AB - Autism is thought to be associated with abnormal hemispheric specialization and left-hemispheric dysfunction. Brain functional imaging using 133Xe-SPECT (single photon emission computed tomography) was used to measure left/right asymmetry and absolute values of regional cerebral blood flow (rCBF) in 18 children with autism aged from four to 17 years and 10 age-matched controls. All controls but only 10 children with autism were right-handed. The left-to-right indices, both hemispheric and regional, were positive in controls, indicating higher left than right rCBF values, but were negative in patients with autism. This inversion was statically significant for total hemispheres, sensorimotor and language-related cortex and was explained by a significant decrease of the left absolute rCBF values in these regions in the patients with autism. The inversion was independent of handedness, sex and age. These results confirm the existence of left-hemispheric dysfunction in childhood autism, especially in the cortical areas devoted to language and handedness, leading to anomalous hemispheric specialization. PMID- 7493720 TI - Effect of early lead exposure on children's postural balance. AB - This study investigated the effect of chronic exposure to lead on children's ability to maintain upright postural balance as a biological marker of lead induced modifications of the neuromotor system. For this study, 162 six-year-old children, with a five-year geometric mean lead concentration in blood of 11.9 micrograms/dL (range 4.0-28.0 micrograms/dL), were tested for postural balance with a microprocessor-based force platform system. An increase in blood lead was significantly associated with an increase in the variable postural sway--implying poorer postural balance. This association was not influenced by socio-economic, racial or environmental factors. This simple, objective and quick technique may be useful for assessing gross motor functions in children who are at or below the United States Centers for Disease Control's class III category and/or for monitoring the effectiveness of medical interventions aimed at reversing lead associated impairment of upright postural balance. PMID- 7493721 TI - International Headache Society classification and diagnostic criteria in children: a proposal for revision. AB - The authors have proposed specific revisions to current International Headache Society (IHS) criteria for the diagnosis of headache. They include: (1) the use of the terms 'clinically definite' and 'clinically probable' to preface the diagnosis of a particular headache disorder to distinguish between situations in which (a) all criteria for the disorder are fulfilled and (b) one or two criteria are not fulfilled; (2) a slight revision to the criteria for the diagnosis of migraine with aura; (3) acceptance by the IHS of a category 'co-existing migraine and tension-type headache'; (4) consistency in the use of terms and elimination of ambiguous phrases; and (5) the omission of rule 7. The authors trust that these suggestions will help make the recommendations of the IHS more applicable to paediatric practice. PMID- 7493722 TI - Locomotion patterns in cerebral palsy syndromes. AB - Locomotion patterns were studied in 160 children with cerebral palsy. Ten patterns were distinguished, which were typical of the type of impairment: (1) crawling--mildly impaired children who will achieve independent walking; (2) creeping and crawling--diplegic children with moderate motor impairment; (3) creeping, never leading to independent walking--children with severe diplegia or tetraplegia; (4) bottom shuffling--children with hemiplegia and ataxia and minimal or mild motor impairment; (5) bunny-hopping--dyskinetic children with marked motor impairment but generally not mentally retarded; (6) rolling- severely diplegic and dyskinetic children; (7) other forms of locomotion, such as bridging or grub-type creeping, rare and typical of children with ataxic elements; (8) just walk--children with hemiplegia, diplegia, ataxic diplegia or ataxia, generally in cases of mild motor impairment but mental retardation; (9) just walk with aids--children with severe diplegia; and (10) no mobility. The locomotion pattern, age at onset and even manner of execution all influenced prognosis for walking. Severe deformity affected the choice of locomotion pattern. Though physiotherapy probably would not greatly influence the adoption of a particular locomotion pattern, early intervention might help prevent deformities. PMID- 7493723 TI - Erythrocyte antioxidant enzyme activities in children with myelomeningocele. AB - Erythrocyte free radical scavenging enzyme activities and their cofactor trace elements in plasma were assessed in 26 selected patients with myelomeningocele, both parents from 10 selected families, and 14 healthy adult controls. All index children except one were deficient in erythrocyte glutathione peroxidase (GSH Px). Nine of 10 parent pairs had at least one parent with deficient GSH-Px activity. Children with myelomeningocele had significantly lower GSH-Px activities than their parents; the group of 10 parent pairs had significantly lower GSH-Px activities than the control group; and glutathione reductase activities were significantly lower in parents and children with myelomeningocele compared with controls. A deficiency in one or more antioxidant enzymes may increase the risk for neural tube defects. PMID- 7493724 TI - Neuropsychological function and MRI abnormalities in neurofibromatosis type 1. AB - This study investigated the relationship between MRI abnormalities and cognitive function in neurofibromatosis type 1.40 children aged eight to 16 years underwent comprehensive neuropsychological, medical and neuroradiological assessments. MRI scans revealed a characteristic pattern of T2-weighted signals ('UBOs') located primarily in the basal ganglia, brainstem and cerebellum in 25 of the children. Reductions in global IQ, attention, and visuopatial and executive functions were shown to occur in association with the presence of UBOs. These findings establish a link between changes in neuropsychological functions and MRI abnormalities in NF-1, and further support neuropathological findings which suggest that UBOs may be a manifestation of delayed or disordered myelination. PMID- 7493725 TI - Neurobehavioral and psychosocial functioning of children with opsoclonus myoclonus syndrome. AB - Childhood opsoclonus-myoclonus syndrome (OMS) is a movement disorder which typically strikes children in the early preschool years, seriously affecting intellectual, social-emotional and general adaptive development. This series of 13 cases with well-documented neurological histories, aged 1.7 to 16.3 years, provides an initial systematic evaluation of these children's neuropsychological, psychosocial and adaptive status. As expected, children with OMS had significantly reduced intelligence and severe speech and motor output problems; however, most of them also demonstrated a range of preserved neurocognitive abilities and impressive goal-directedness and communicative effort. Psychosocial problems included mild behavioral impairment on the Achenbach Child Behavior Checklist, and severe adaptive limitations on the Vineland Adaptive Behavior Scales despite relatively strong social skills. Developmental factors and likelihood of subcortical localization are discussed, and practical guidelines are provided for behavioral and educational management of these children. PMID- 7493726 TI - Bilateral painful epileptic seizures of the hands. AB - Secondary sensory seizures (SSS) are very rare epileptic seizures. A 14-year-old male patient with habitual painful seizures of the backs of both hands since the age of four is reported. The painful seizures were rarely followed by impairment of consciousness. Ictal EEG showed 4 to 6Hz rhythmic waves over the right anterior and mid-temporal area following complaints of pain. The absence of tumorous lesions on brain CT or MRI, childhood onset and the absence of combined seizures were characteristic features of this case compared with other patients reported to have SSS. PMID- 7493727 TI - [Familial poisoning with Karwinskia humboldtiana]. AB - The ingestion of ripe fruit of the Karwinskia humboldtiana, a shrub commonly known as tullidora or coyotillo, produces an intoxication described in the literature as a symmetric flaccid paralysis of the hind limbs, progressive and ascendent, that in severe cases may cause bulbar paralysis and death. The cause of an acute accidental intoxication of an entire family is presented here, wherein ten out of thirteen members ingested the ripe fruit of the tullidora. Three died, the father and two daughters. For the first time the toxins determination in blood by thin layer chromatography method is described. This method supports the diagnosis with other polyradiculoneuritis such as poliomyelitis and the Guillain Barre's syndrome. PMID- 7493728 TI - [Diagnosis: plasmocytoma]. PMID- 7493729 TI - [Advances of molecular genetics in the study of diabetes]. PMID- 7493730 TI - [New approaches in the treatment of arterial hypertension]. PMID- 7493731 TI - [Poliomyelitis and acute flaccid paralysis. Perspectives for its epidemiologic surveillance]. PMID- 7493732 TI - [The free-trade treaty and health services]. PMID- 7493733 TI - [The physician, ethics, and the university]. PMID- 7493734 TI - [Diagnostic strategies]. PMID- 7493735 TI - [Phosphatidylcholine induces an increase in the production of interleukin-6 and improves survival of rats with neonatal sepsis caused by Klebsiella pneumoniae]. AB - Infections by gram-negative bacteria are one of the major causes of death in newborns. Bacterial clearance is deficient in septic neonates, which seems to increase their susceptibility to infections. In this study, we observed a significant improvement in clearance of Klebsiella pneumoniae in newborn wistar rats inoculated by intraperitoneal via with 800 mg k soybean phosphatidylcholine (PC), compared to the control group injected with PBS (p 0.05). The overall survival rate was improved (p 0.05) and the white blood cell counts showed a greater leukocytosis and neutrophilia during the peak of bacteremia in the PC treated animals. Circulating levels of interleukin-6 were greater in the PC group, which developed an intense splenic hematopoiesis of the granulocyte (p 0.05) and megakariocyte series (p 0.01). No significant changes were observed in bone marrow granulocyte deposits in both study groups. The improvement in survival rate, the changes in leukocyte counts and the splenic hematopoiesis may be associated with the increased production of IL-6. These results suggest that IL-6 plays a role in the protection mechanism induced by PC in this experimental model of newborn septicemia. PC seems to be an immunomodulator of the acute response to gram-negative bacterial infection. PMID- 7493736 TI - [Characterization of surface antigens of the nematode parasite Trichinella spiralis: study of its role in protection mechanisms and their usefulness in the diagnosis of trichinosis]. AB - Among the most important aspects in the study of trichinosis are the development of specific and sensitive diagnostic methods for the detection of the infection by the parasite as well as the characterization of antigens from Trichinella spiralis that induce protection in the host. In the context, the characterization of surface stichosome and excretory secretory antigens of T. spiralis muscle larvae has been an important issue due to the high immunogenicity of such components in most hosts so far studied. In this work, we have been able to identify and characterize molecules from both compartments of muscle larvae. These components have been used in assays for specific detection of T. spiralis infections particularly in pigs, as well as in assays to evaluate their role in the induction of protection in mice. PMID- 7493737 TI - [The "Fantasy Island" syndrome. Identification of a new osteochondrodysplasia probably of autosomal dominant type]. AB - Clinically, two unrelated patients, an adult male and a female child, coming from non-consanguineous parents, presented dwarfism, peculiar facies, with blepharophimosis, mongoloid slanted eyes, abundant eyebrows and eyelashes, harsh voice and short hands and feet. Radiologically, they presented brachymetacarpalia, brachymetatarsalia, and brachyphalangia of all fingers and toes, shortened and broadened long bones with normal morphology, hypoplastic pelvis and shape anomalies of the vertebral bodies. The clinical and radiological concordance and the differential diagnosis, mainly with acromicric dysplasia, allow the characterization of a distinctive osteochondrodysplasia probably due to an autosomal dominant mutation. PMID- 7493738 TI - [Mexican phenotype and genotype Vibrio cholerae 01]. AB - This paper presents the phenotypical and genotypical characterization of 26922 Vibrio cholerae 01 strains isolated in Mexico from 1991 to 1993. All strains isolated were El Tor biovar. Strains were sensitive to antibiotics excluding furazolidone, streptomycin and sulfisoxasole to which we found resistance in 97% and we are using this characteristic as epidemiological markers. We detected a marked change in frequency of Inaba serotype from 1991, when it was dominant, with 99.5%, until 1992 when Ogawa serotype turned to be dominant with 95% of isolates. All Vibrio cholerae 01 strains, except one Ogawa strain, were to igenic, and V. choleraeno 01 were not toxigenic by ELISA, PCR and cell culture tests. Dominant ribotype was 5, but we found some strains with 6a pattern and two with ribotype 12. We are searching for ribotype 2 among hemolytic strains in order to learn if there is any relation to Gulf Coast strains prevalent in the USA, but until now we have not found any V. cholerae ribotype 2 in our isolates. Even if rapid tests are recommended for immediate diagnosis of cholera, it is necessary to continue bacterial isolation in order to have strains for phenotyping and genotyping studies that may support epidemiological analysis. PMID- 7493739 TI - [Dermatology topics]. PMID- 7493740 TI - [The good use of experimental animals]. PMID- 7493741 TI - [Alzheimer's disease. Considerations for its clinical diagnosis]. AB - This paper present features for identifying, on a clinical basis, the syndromatic complex "primary degenerative dementia of Alzheimer's disease type". Data are included which aid in the identification among the broad picture of dementia disorders. Information is presented about the most recent histopathological findings, the study proceedings and therapeutic trends. PMID- 7493744 TI - [Reflections on Mexican medicine]. PMID- 7493742 TI - [Evolution and current status of certification of specialists in Mexico]. PMID- 7493743 TI - [Clinical engineering. Rationale for implementing the technology in hospitals]. AB - A new clinical engineering service was started in the Hospital General de Mexico, almost 5 years ago, with the technical assistance of Universidad Autonoma Metropolitana and financial support of Patronato del Hospital General de Mexico, A.C. The purpose of such service is to improve all aspects of medical device management in the hospital, i.e., selection, acquisition, installation, quality control, use and maintenance. The project also includes education, research and development related to health care technology. Interesting results have been obtained in each of the areas of clinical engineering, even during the early stages. They are herein presented to the medical community. PMID- 7493745 TI - Depression in long-term care facilities. AB - Depression occurs in 15 to 25% of elderly patients in long-term care settings. It is frequently misdiagnosed and inadequately treated. However, given the significant morbidity and mortality associated with late-life depression, the clinician needs to be aware of it and treat it aggressively with appropriate pharmacologic and psychotherapeutic interventions. PMID- 7493746 TI - The primary care setting: managing medical comorbidity in the elderly depressed patient. PMID- 7493747 TI - Principles to optimize drug treatment in the depressed elderly: practical pharmacokinetics and drug interactions. PMID- 7493748 TI - Depression in the elderly. PMID- 7493749 TI - Rational antidepressant selection in the elderly. AB - Proper selection of antidepressant therapy is of growing importance as new and unique agents are added to the therapeutic armamentarium. Physicians must expend some effort in becoming familiar with the current treatment choices to facilitate selecting agents tailored to the needs of individual patients. The criteria that should be considered in antidepressant selection, assuming equal efficacy at appropriate therapeutic dosages, include propensity of a drug to produce side effects, patient and family history of response to previous antidepressant therapy, pharmacokinetics, ease of administration, drug interaction potential, and emerging data that suggest differential efficacy in specific depressive syndromes. PMID- 7493750 TI - Recognition and differentiation of elderly depression in the clinical setting. AB - Primary care physicians must realize that depression is a prevalent source of excess disability among their elderly patients. However, when depression strikes the elderly patient, it often goes unrecognized and untreated. By maintaining a high index of suspicion, clinicians will be prepared to conduct a focused clinical interview that can uncover depression otherwise masked by prominent somatic and anxiety-related complaints. Initiation of antidepressant treatment in elderly depressed patients is an important life-enhancing and life-saving measure. PMID- 7493751 TI - Induction of lytic pathways in T cell clones derived from wild-type or protein tyrosine kinase Fyn mutant mice. AB - The OVA-reactive CD4+ Th1 clones and alloreactive CD8+ clones derived from wild type or fyn-/- mice serve as model systems which have allowed us to investigate several aspects of the molecular events associated with T cell-mediated cytotoxicity, including 1) the differential utilization of two distinct cytolytic pathways by CD4+ Th1 clones and CD8+ CTL, 2) a comparison of the pathways of lysis induced by stimulation of the TCR or by alternative stimuli, 3) the requirement of Fyn for derivation of antigen-specific T-cell clones having properties of CD4+ Th1 and CD8+ CTL cells 4) the differential requirement of Fyn in the induction of responses by TCR and the alternative stimuli. Stimulation through the TCR, either by APC bearing relevant antigen or by immobilized anti CD3 mAb, resulted in comparable levels of target cell lysis by clones from both wild-type and fyn-/- mice. These clones also utilize the Fas pathway to lyse target cells. Thus, Fyn does not appear to be required for expression of the Fas pathway when triggered through the TCR. In contrast, lysis of target cells by T cell clones lacking Fyn was deficient when stimulated through Thy-1 or Ly-6C (using mAb) or with Con A or phorbol ester as compared to clones derived from wild-type mice. The basis for the defect in response to stimulation through the GPI-linked molecules appears to be a signaling defect which affects all of the functional responses we measured, while the defect in response to Con A stimulation appears to affect lysis but not lymphokine production. Thus, Fyn expression is selectively required for efficient activation of the Fas pathway of lysis through Thy-1, Ly-6C, and by Con A or phorbol ester in these T-cell clones. CD8+ clones derived from fyn-/- mutant mice, like clones derived from wild-type mice, display antigen-specific lysis, and appear to express perforin message and perforin protein. A Ca(++)-dependent (presumably perforin/exocytosis) component and Fas component of lysis was detected in CD8+ clones derived from fyn-/- mutant mice. Thus, Fyn is not required for expression of these components of antigen specific lysis by CD8+ alloreactive CTL clones. It appears that CD8+ clones that use multiple lytic mechanisms may selectively employ the perforin or Fas-based pathway depending on properties of the target cell or stimulus.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 7493753 TI - Molecular interactions in the activation of effector and precursor cytotoxic T lymphocytes. AB - Cell-cell interactions are influenced by parameters that cannot readily be studied using either intact cells or soluble molecules. Replacing one of the pair of interacting cells with an artificial cell surface construct allows novel insights to be gained into some of these parameters. Application of this approach to the study of CTL has helped to clarify the contrasting roles of some of the various receptors that are involved in recognition, adhesion and activation. In addition, it has revealed features of these receptor ligand interactions that help to explain how CTL are able to carry out effective immune surveillance and elimination of virus-infected or tumor cells. Although not discussed in this review, artificial cell surface constructs have also been effectively employed to study the interaction of TH cells with class II bearing surfaces. Class I protein and peptide antigen can be sufficient to mediate adhesion and activate CTL effector function through the TCR and CD8. In addition, interactions of other co receptors with their ligands can act along with TCR and CD8 in a cascade of activated adhesion and co-stimulatory signal generation to allow adhesion and response when antigen and/or class I surface densities are too low to be sufficient by themselves to initiate response. The relative contributions of the various receptor/ligand interactions to a given CTL/target encounter will depend upon the affinity of the TCR for antigen and on the densities and types of ligands, including antigen, displayed on the target cell surface. It appears that the CTL has the ability to accomplish its task in a variety of ways, providing it with considerable flexibility in recognizing and eliminating antigen-bearing target cells. Thus, downregulation of any one particular ligand on a virus infected or tumor cell does not allow escape from CTL surveillance provided that at least a low level of class I antigen remains present. The CTL is able to employ several co-receptors specific for ligands common to many cell types without being diverted from effective immune surveillance, since these receptors only become activated to mediate high-avidity adhesion when antigen is detected by the TCR. Cloned effector CTL are most amenable to studies of the kind reviewed here, since large numbers of homogenous cells can be obtained, antigen-specific adhesion can be readily measured and response is rapid and easily quantitated.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 7493752 TI - Perforin and lymphocyte-mediated cytolysis. AB - We have discussed in the previous sections the recent progress made toward elucidating the regulatory mechanism of perforin gene transcription and the domain structure of the perforin molecule. It appears that the expression of perforin is, at least partially, controlled at the transcription level through the interaction between killer cell-specific cis- and trans- acting factors. One of such cognate pairs, NF-P motif (an EBS-homologous motif) and NF-P2 (a killer cell-specific DNA-binding protein), has been described. The regulatory mechanism of gene transcription, however, is likely to involve multiple factors which act in a coordinated fashion to bring about the most efficient expression of perforin limited strictly to activated killer lymphocytes. Through studies using synthetic peptides and recombinant perforins, it has been suggested that the N-terminal region of the perforin molecule is an important, though not the only, domain responsible for the lytic activity. Further studies are warranted to elucidate the role(s) of other potential amphiphilic structures located in the central portion of the perforin molecule in the overall pore-forming activity. The molecular basis underlying the resistance of killer lymphocytes to perforin mediated lysis still remains an open question. Preliminary results, however, suggest that the surface protein(s) restricted to killer cells may account for their self-protection against perforin. Based on recent studies using perforin deficient mice, the involvement of perforin in lymphocyte-mediated cytolysis both in vivo and in vitro has been confirmed. Two functional roles, a direct (lytic) and an indirect (endocytosis enhancer; conduit), both of which may contribute critically to the cell-killing event can be attributed to perforin. The fact that lymphocytes may also employ perforin-independent killing mechanism(s), e.g. Fas dependent pathway, is beyond the scope of this review. There is, nevertheless, no doubt that these alternative cytolytic mechanisms may also play important roles in immune effector and/or regulatory responses associated with killer lymphocytes. Obviously, we are still a long way from concluding on the functional relevance of each individual cytolytic mechanism seen in different physiopathological situations. Suffice it to say, however, that a wealth of information on lymphocyte-mediated killing has already emerged through the multidisciplinary efforts conducted in our and other laboratories that promise to further dissect this complicated event in the years to come. PMID- 7493754 TI - PELs and the perforin and granzyme independent mechanism of CTL-mediated lysis. AB - The central role of CTLs in immunopathology accounts for the increasing interest in deciphering the mechanism whereby they kill at the molecular level. Recent studies show that CTLs have two molecularly distinct lytic mechanisms at their disposal. The first involves the direct effect(s) of the pore-forming protein perforin, possibly in conjunction with granzymes. In recent years, experiments conducted in our laboratory led to an alternative pathway, of receptor-mediated mechanism for CTL killing, involving neither the secretion nor the lytic action of the pore-forming protein perforin or of granzymes. By this mechanism, engagement of a CTL membrane ligand and an apoptosis-inducing target cell surface receptor triggers the disintegration of the CTL-bound target cell. Cross-linking of apoptosis-inducing target cell surface molecules (e.g. Fas), induced upon binding of CTL ligands (e.g. Fas-L), may be required and sufficient to trigger target cell apoptosis. Intracellular lethal signals emanating from the cross linked intracellular death domain of Fas are postulated. PMID- 7493755 TI - Granzyme B plays a critical role in cytotoxic lymphocyte-induced apoptosis. PMID- 7493756 TI - Fas-mediated cytotoxicity--a new immunoregulatory and pathogenic function of Th1 CD4+ T cells. PMID- 7493757 TI - Nuclear changes in the cytotoxic T lymphocyte-induced model of apoptosis. PMID- 7493758 TI - Molecular pathways of CTL-mediated cytotoxicity. PMID- 7493759 TI - Fas bridging cell death and cytotoxicity: the reaper connection. PMID- 7493760 TI - Role of extracellular ATP and P1 and P2 classes of purinergic receptors in T-cell development and cytotoxic T lymphocyte effector functions. PMID- 7493761 TI - Mechanism and biological significance of CD4-mediated cytotoxicity. AB - It is now well established that CD4+ T cells can express cytotoxic activity. This type of cell-mediated cytotoxicity is associated with the Th1-, but not with the Th2-phenotype. While the activation of CD4+ CTL is MHC class II-restricted, the effector phase, i.e. the target cell killing is unrestricted and antigen non specific. In analogy to CD8+ CTL, CD4-mediated target cell death is by DNA fragmentation. However, the molecular mechanism of killing differs from CD8 mediated lysis. Thus, CD4+ CTL preferentially lyse their targets via Fas-Fas ligand interaction, whereas the major cytotoxic effect of CD8+ CTL is by granule exocytosis, i.e. perforin and granzymes. Although CD8+ CTL can also express the FasL, their lytic activity through interaction with Fas is of less importance. Likewise, some CD4+ CTL may also kill by perforin/granzymes activity, but this pathway is of minor significance. The aims of CD8- or CD4-mediated lysis are also different. Thus, the major task of CD8+ CTL which recognize and kill their targets in the context of MHC class I molecules, is the lysis of virally infected cells and battling against tumor cells. CD4+ CTL, on the other hand, have an immunomodulatory role. Thus, they preferentially eliminate activated MHC class II positive cells, i.e. APC, be they monocytes/macrophages, B cells or T cells. They may lyse these cells in order to prevent an overreaction of the ongoing immune response or in order to remove potentially hazardous cells upon completion of the immune response. The Fas-FasL pathway is particularly suitable for this task as myeloid or lymphoid cells express Fas only if activated, while FasL is preferentially expressed on activated CD4+ Th1 cells. Moreover, activated T cells eliminate themselves by the Fas-mediated pathway. Whether this happens by fratricide only, or also by suicide or both is open. Moreover, CD4+ CTL are particularly suitable for killing tumor cells as well, as they are efficient effectors in bystander lysis in contrast to CD8+ CTL. On the other hand, the non specific killing via Fas-FasL interaction, which is an important reason for the bystander lysis, may have unwanted effects in that cells which should not be eliminated could be killed. Such reactions affecting various organs and cells, e.g. the liver, thyroid or islet cells of the pancreas could be an explanation for certain autoimmune diseases. PMID- 7493762 TI - Expression of CD11a and CD45R isoforms defines distinct subsets of CD8+ TCR alpha beta and TCR gamma delta CTL in vivo. AB - Intraperitoneal immunization of rats with a syngeneic lymphoma and allogeneic leukocytes induced enrichment of antigen-selective TCR alpha beta+ and TCR gamma delta+ CTL. The peritoneal cavity seems to be a suitable site for enrichment of antigen-selective CTL, since the induced effector cells executed strong cytotoxicity without any requirement for in vitro reactivation. Tumor-selective CTL expressed high cell surface levels of CD45RC, allogeneic CTL expressed a variable level of CD45RC, while SAg-reactive CTL had low CD45RC expression. CTL with tumor and allogeneic selectivity as well as SAg-induced CTL all expressed the LFA-1high phenotype, suggesting that upregulation of LFA-1 is a hallmark for in vivo activated CTL. RT-PCR analyses showed that all CD8+TCR alpha beta+ CTL lost expression of CD45R exon 4 mRNA, which is compatible with the view that effector/memory T cells are CD45RA-. In contrast, TCR gamma delta+ CTL retained the CD45RA phenotype but showed a down-regulation of CD45R exon 8 mRNA. Since isoforms of the CD45 tyrosine phosphatase have been reported to differentially affect T-cell activation, the unique CD45R splice pattern observed in TCR alpha beta+ and TCR gamma delta+ CTL implies that CD45R may influence the TCR signal transduction distinctly in various effector CTL subsets. PMID- 7493763 TI - Lymphocyte-mediated cytotoxicity in vitro and in vivo: mechanisms and significance. PMID- 7493764 TI - Tissue expression, structure and function of the murine Ly-6 family of molecules. AB - Murine Ly-6 molecules are a family of cell surface glycoproteins which have interesting patterns of tissue expression during haematopoiesis from multipotential stem cells to lineage committed precursor cells, and on specific leucocyte subpopulations in the peripheral lymphoid tissues. These interesting patterns of tissue expression suggest an intimate association between the regulation of Ly-6 expression and the development and homeostasis of the immune system. Ly-6 molecules are low molecular weight phosphatidyl inositol anchored glycoproteins with remarkable amino acid homology throughout a distinctive cysteine rich protein domain that is associated predominantly with O-linked carbohydrate. These molecules are encoded by multiple tightly linked genes located on Chr. 15 which have conserved geneomic organization. The in vivo functions of Ly-6 molecules are not known although in vitro studies suggest a role in cellular activation. This review will summarize our understanding of Ly-6 with regard to tissue expression, molecular structure, gene organization and function. PMID- 7493765 TI - T cell receptor usage in autoimmune disease. AB - Activated T-cells are believed to play a critical role in the pathogenesis of autoimmune disease. In experimental allergic encephalomyelitis (EAE), an animal model resembling human multiple sclerosis (MS), there is evidence that T cells reactive to myelin basic protein mediate an inflammatory response within the central nervous system leading to demyelination. Furthermore, encephalitogenic T cells express TCR with highly restricted V gene usage and consequently specific forms of immunotherapy directed against V gene products have been successful in preventing and treating EAE. These findings prompted studies into the analysis of TCR repertoire expression in human autoimmune diseases in an attempt to identify the TCR usage of autoreactive and potentially pathogenic T cells. However, this has proved difficult as the autoantigens that drive the T cell response in most human autoimmune disorders are unknown. This review examines the data that have accumulated over the past few years on TCR usage in human autoimmune diseases and is focused largely on rheumatoid arthritis and MS. PMID- 7493767 TI - Cell mediated immunity in Antarctic wintering personnel; 1984-1992. AB - Cell-mediated immune responses were studied in 12 Antarctic and sub-Antarctic wintering groups at quarterly intervals over the period 1984-1992, using the cutaneous CMI Multitest. These populations are among the most isolated on earth. While the sub-Antarctic population at Macquarie Island had levels of responsiveness and hypoergy (9%) comparable to healthy populations in temperate zones, the Antarctic Continental group showed a level of hypoergy of 36%. There was no seasonal variation in the pattern of responses. It is concluded that the extreme and isolated environment and stress factors are responsible for the decreased immunological responsiveness but the mechanisms are presently unclear. On review, one factor appears to be perceived anxiety. The high rate of hypoergy in Antarctica, where medical care is limited, may have health implications. These groups provide an excellent analogue for immunological investigations in longer term space flight. PMID- 7493766 TI - MHC proteins and heparan sulphate proteoglycans regulate murine cytomegalovirus infection. AB - Factors influencing MCMV infection mediated by MHC class 1 molecules were analysed further as previous studies showed that the effects of the MHC genotype on sensitivity to this virus are important in vivo. Here we show that H-2d, H-2b, H-2r and H-2v macrophages are highly sensitive to MCMV. Moreover, transfection of H-2k L-cells with Kb or Dd conferred sensitivity to MCMV. This was not affected by amino acid substitutions in Kb alpha 1 or alpha 2, although previous studies demonstrated that exchange of the alpha 1 domain of Dd with Ld alpha 1 compromised sensitivity. Here replacement of Kb alpha 3 with Ld alpha 3 reduced susceptibility to low doses of MCMV. In addition, extracellular beta 2 microglobulin (beta 2m) promoted infection of beta 2m-negative RIE/TL8X.1 cells transfected with Db with or without a beta 2m gene. Hence MCMV infection can involve beta 2m and the alpha 1 and alpha 3 domains of MHC heavy chains. MCMV infection of L-cells expressing Dd or Kb was also inhibited by heparin, but infection of the parental L-cell line was not reproducibly affected. A role for heparan sulphate proteoglycan in MHC-mediated MCMV infection was confirmed using cells pre-treated with heparinase I or III, or propagated in chlorate to inhibit the sulphation of the glycosaminoglycan chains. PMID- 7493769 TI - Assessment of the therapeutic potential of cytokines, cytotoxic drugs and effector cell populations for the treatment of multiple myeloma using the 5T33 murine myeloma model. AB - The therapeutic potential of six cytokines, eight cytotoxic drugs and two effector cell populations for the treatment of multiple myeloma was assessed in vitro using the 5T33 murine myeloma model. The efficacy of combination IFN-alpha and melphalan therapy was also evaluated in vitro and in vivo. Of the cytokines tested in vitro using the MTT assay, only IFN-alpha demonstrated significant inhibition of myeloma cell growth at non-toxic concentrations (ED50 = 1508.3 +/- 181.3 U/mL and 2617.9 +/- 334.0 U/mL for murine IFN-alpha [mIFN-alpha] and human IFN-alpha hybrid B/D [hIFN-alpha B/D], respectively). The ED50 for the eight cytotoxic drugs tested ranged from 2.3 x 10(-9) to 4.3 x 10(-13) mol/L and all were within the therapeutic range for humans. Combination hIFN-alpha B/D and melphalan were found to be additive in their inhibitory effects on myeloma cell growth in vitro and this finding was confirmed in vivo in C57BL/KaLwRij mice bearing disseminated 5T33 myeloma. Control animals demonstrated a median survival duration of 25.3 days whereas hIFN-alpha B/D or melphalan treatment alone increased survival to 30.5 and 33.3 days, respectively (P < 0.001). Combination IFN-alpha/melphalan therapy increased median survival duration to 38.5 days (P < 0.001) which was also significantly greater than that obtained with single agent therapy (P < 0.01). The murine myeloma cells were found to be resistant to NK cell lysis but susceptible to lysis by LAK cells (49.3 +/- 6.3% lysis at an effector to target ratio of 100:1).(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7493770 TI - Establishment of a natural suppressor cell line producing soluble suppressor factor other than transforming growth factor-beta. AB - Natural suppressor (NS) cell line (Clone 59) was established from the bone marrow of adult C3H/Hej mice in the presence of WEHI-3 conditioned media. Clone 59 cells suppressed the generation of cytotoxic T lymphocytes from normal mouse spleen. This suppression was seen at a responder-to-suppressor cell ratio of 1000:1 and lacked antigen specificity or MHC restriction. Clone 59 cells expressed the 'null' surface phenotype (Thy1.2-, CD3-, Lyt-2-, L3T4-, surface Ig-, MAC-1-) by immunofluorescent staining. Clone 59 cells exhibited no cytolytic activity against NK cell-sensitive YAC-1 and natural cytotoxic L929 target cell lines. Non specific suppression, with a cell-free supernatant from the Clone 59-NS cells, also was observed. The supernatant did not inhibit [3H]-thymidine uptake by CTLL 2 cells which were proliferating in response to IL-2. Anti-transforming growth factor-beta (TGF-beta) monoclonal antibody had no effect on suppression, suggesting that the non-specific suppression is mediated by some soluble factors other than TGF-beta. Clone 59 cells may be useful in identifying non-specific suppressor cells in adult bone marrow and studying their functional role in the regulation of tolerance and self-reactivity. PMID- 7493771 TI - Co-operative effect between insulin-like growth factor-1 and interleukin-2 on DNA synthesis and interleukin-2 receptor-alpha chain expression in human lymphocytes. AB - Recently we demonstrated that IGF-1 has the same effect on DNA synthesis and G1 phase length during human lymphocyte proliferation as IL-2. In order to determine the link between IGF-1 and IL-2 on lymphocyte proliferation, we tested the ability of these cytokines, alone or in combination, to induce DNA synthesis and increase the number of cells expressing IL-2-alpha chain receptors. Our results showed that the increase in DNA synthesis produced by the addition of IGF-1 to cells cultured with saturable concentrations of IL-2, correlated well with an increase in the number of cells expressing IL-2-alpha chain receptor. The effect on both DNA synthesis and IL-2-alpha chain receptor expression was greater at lower concentrations of PHA. This study suggests that the proliferative effect of IGF-1 might be due to separate stimulation of different cell populations and/or by activation of a tyrosine-kinase signal transduction pathway complementary to the IL-2 activation signal. PMID- 7493768 TI - Immunohistological staining of lymphoid tissue in four Australian marsupial species using species cross-reactive antibodies. AB - Antibodies raised against evolutionarily conserved intracytoplasmic peptide sequences of human T and B lymphocyte antigens and an antibody raised specifically against koala serum IgG were assessed for their capacity for immunoperoxidase staining of formalin fixed lymphoid tissues of koalas (Phascolarctos cinereus), common brushtail possums (Trichosurus vulpecula), common ringtail possums (Pseudocheirus peregrinus) and tammar wallabies (Macropus eugenii). Utilizing microwave pretreatment and a streptavidin biotin-horseradish peroxidase method for histochemical staining, polyclonal anti-human CD3 (for T cells), monoclonal anti-human CD5 (for T cells), monoclonal anti-human CD79b cy (for B cells) and polyclonal anti-koala IgG (for plasma cells) consistently stained lymphoid elements in a wide range of tissues from all four marsupial species. Monoclonal anti-human CD79a cy (for B cells) stained lymphoid elements in only brushtail possums. This is the first report of immunohistological staining of lymphocytes in Australian marsupials, thereby vastly increasing the scope for immunological studies in these species. PMID- 7493773 TI - Deficiency in T cell responses of human fetal lymph node cells: a lack of accessory cells. AB - Lymph node and spleen cells of human foetuses from the 18th to the 24th week of gestation were analysed with regard to their phenotypes and their functional capacities. Fetal mesenteric lymph nodes contain high percentages of CD45RA+ T cells and few B cells and monocytes, whereas the fetal spleen is comprised of equal numbers of T and B cells as well as monocytes/macrophages. Functional analysis of lymph node T cells revealed a lack of proliferative response to PHA or CD3 specific mAb, despite induction of expression of the activation marker CD69. Proliferation of LN cells and thymocytes was observed upon addition of exogenous IL-2. An allogeneic EBV transformed tumour cell line, known to be an effective antigen presenting cell, could induce proliferation of LN cells without exogenous IL-2 and fetal spleen cells could proliferate in response to all stimuli tested without additional IL-2. Splenic non-T cells could restore the proliferation of lymph node cells as efficiently as IL-2 or the EBV transformed B cell line. Separated B cells were more effective than plastic adherent cells on a per cell basis. Naivity of the fetal immune system is therefore not only reflected by the expression of markers representative for naive lymphocytes but can also be due to the absence of potential accessory cells in the different lymphoid organs. PMID- 7493774 TI - Cytokine mRNA expressed in tuberculin skin test biopsies from BCG-vaccinated and Mycobacterium bovis inoculated cattle. AB - To obtain a better understanding of the delayed-type hypersensitivity reaction to Mycobacterium bovis, we measured the expression of cytokine mRNA from tuberculin skin test biopsies of cattle. Non-vaccinated and BCG-vaccinated cattle were inoculated intratracheally with a low dose of virulent M. bovis or sham inoculated and 20 weeks later were skin tested with tuberculin. At necropsy 1-2 weeks later, tuberculous lesions were found in six of the nine non-vaccinated and three of the nine BCG-vaccinated animals. All of the lesioned and the majority of the non-lesioned M. bovis inoculated cattle showed a distinct skin swelling response to tuberculin, irrespective of vaccination. However, cattle with tuberculous lesions displayed larger skin swelling responses than non-lesioned cattle. Tuberculin-induced expression of IFN-gamma, IL2, IL4, IL10 and TNF-alpha mRNA occurred in the skin biopsies of all of the lesioned, M. bovis inoculated animals except for an absence of tuberculin-induced TNF-alpha mRNA expression in two animals. A lower proportion of the non-lesioned M. bovis inoculated cattle displayed tuberculin-induced expression of the five cytokine mRNA. There was no evidence of a unique pattern of cytokine expression which could be used to distinguish between diseased and protected animals. By 28 weeks after vaccination, the three BCG-vaccinated, sham-inoculated cattle displayed minimal skin swelling response to tuberculin, but tuberculin-induced expression of IFN gamma, IL2, IL4, IL10 and TNF-alpha mRNA was observed in skin biopsies of all of these animals.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7493772 TI - Immune response to hepatitis C virus core protein in mice. AB - To analyse the immune response to the hepatitis C virus (HCV) core protein, we immunized mice with the protein. BALB/c (H-2d) and C3H/He (H-2k) mice were high responders, while C57BL/6 (H-2b) mice were low responders in terms of Th cell proliferative responses. All the strains showed comparable levels of antibody responses to the HCV core protein. The Th cell lines recognized residues 61-90 of the HCV core protein in the context of I-Ad (BALB/c) and residues 11-30 in the context of I-Ek (C3H/He), respectively. The Th cell lines were restricted by I-Ab in C57BL/6 mice but recognized no synthetic peptide that spanned the region, although derivative clones from the line recognized residues 1-20 and 91-110 of the HCV core protein, respectively. The Th cell lines were Th 1 subset in all three strains based on the profile of lymphokine secretion. The major B cell epitope of the protein was found to be within residues 21-40 of the HCV core protein in all three strains. These observations should be useful for better understanding of the immune response to the HCV core protein in vivo. PMID- 7493775 TI - Search for linkage disequilibrium between alleles in the T cell receptor alpha and beta chain loci and susceptibility to rheumatic fever. AB - A population-based linkage disequilibrium study was conducted to search for associations between alleles in T cell receptor alpha and beta chain polymorphic loci and susceptibility to rheumatic fever. The allele frequencies of four T cell receptor locus restriction fragment length polymorphisms were measured in 47 European and 51 Maori subjects with a history of rheumatic fever. These allele frequencies were compared to the allele frequencies in three or four independently recruited, race-matched control groups totalling 125 Europeans and 117 Maoris with no history of rheumatic fever. The polymorphisms studied were (locus/enzyme/probe) C alpha/Taq1/Y14, V alpha/Taq1/Y14, V beta 7/BAMHI/V beta 7.4 and V beta 8/BAMHI/V beta 8.1. There was no evidence for linkage disequilibrium between rheumatic fever and these Tcr alleles in either the Maori or European subjects. PMID- 7493776 TI - The 65 kDa protein of Mycobacterium habana and its putative role in immunity against experimental tuberculosis. AB - Mycobacteria including Mycobacterium tuberculosis and Mycobacterium leprae possess multiple antigens some of which inhibit other anti-mycobacterial immune responses. Whole cell vaccines are not free from these suppressive molecules and may adversely affect the immunogenic response(s). Purified protein components having only immunogenic properties should prove to be superior vaccine(s). Mycobacterium habana, a candidate vaccine for mycobacterial infections has been dissected for analysing its antigenic myriad. A 65 kDa protein of this mycobacterium has been isolated and characterized for its protective and cell mediated immune responses. The protein was isolated in pure form using an isotachophoresis (SDS-PAGE filtration) technique and identified with low molecular weight markers along with mAb using the immunoblot technique. Mab IIH9 has identified a 65 kDa protein in M. habana. This protein has been found to be immunoprotective in mice against M. tuberculosis H37Rv infection. It generates high levels of DTH responses in mice against M. tuberculosis and M. leprae antigens and inhibits migration of sensitized cells under the antigenic influence of homologous and heterologous origin. Possibilities of developing this protein as a subunit vaccine are discussed in this report. PMID- 7493777 TI - The immune system and integrated homeostasis. AB - The immune system is usually viewed as an effector mechanism reacting to environmental antigenic challenge with defensive responses designed to eliminate 'foreign' substances and return to standby or surveillance mode. However, there is now substantial evidence to suggest that immunity is not effector-biased but is also a sensory organ and forms part of an integrated homeostatic network. Evidence that communication pathways exist between the immune and central nervous systems which support bi-directional information flow, and that the output of the immune system influences other physiological adjustments to environmental change, is discussed in this paper. The proposition that an endocrine-immune gradient determines the homeostatic response to combined microbial and other ambient stressors is established and the potential for management of this gradient is explored. PMID- 7493778 TI - Audit--guidelines, policies, best practice or 'cookbook medicine'? PMID- 7493779 TI - Cardiac troponin T release in multiply injured patients. AB - Cardiac troponin T (cTnT), a new marker of myocardial tissue damage, was investigated in 32 consecutive multiply injured patients. cTnT, creatine kinase (CK) and CK isoenzyme MB (CK-MB) mass concentrations were measured immediately after admission, 12 and 24 h later and daily thereafter for 4 days. We found a moderate increase in cTnT in 22 patients (72 per cent; peaks: 0.6-5.1 micrograms/l). In only four of these 22 patients did the CK-MB mass/CK index indicate myocardial injury. ST-T alterations and arrhythmias did not occur significantly more frequently in patients with increased cTnT plasma concentrations or positive CK-MB mass/CK index. We found a moderate increase in cTnT in 72 per cent of all patients with multiple injuries, but we found no association between an increase in cTnT and the occurrence of electrocardiographic changes and arrhythmias. PMID- 7493780 TI - Finite element analysis of the stress distributions in the proximal end of the femur after stabilization of a pertrochanteric model fracture: a comparison of two implants. AB - The study investigated the influence of two different implants--the dynamic hip screw (AOR) 135 degrees with 2-hole-plate and the 135 degrees gamma-nail (HowmedicaR)--on the resulting stress and strain distributions in the proximal femur after stabilization of an idealized trochanteric fracture under typical physiological loading conditions. Data recorded for the uninjured femur were used for comparison. Two-dimensional plane stress finite element models of the femur and the implants were used for the calculations. The three-dimensional nature of this problem was approximated by an appropriate distribution of element thickness according to the real geometry in combination with an overlay technique, superimposing special finite element layers for each constituent of the bone implant system (i.e. steel, cortical and cancellous bone). Three variations of the idealized fracture were introduced, i.e. free, with callous and bony bridging, the screw being modelled in a movable ('dynamic') as well as in a locked state. As far as the resulting stress distributions are concerned, neither of the two implants proved to be significantly superior to the other for stabilization of the idealized fracture selected. PMID- 7493781 TI - Some basic biomechanical characteristics of medullary pressure generation during reaming of the femur. AB - We measured femoral intramedullary pressures and applied axial thrust force generated in vitro during reaming with the AO and Zimmer systems. Six pairs of cadaver femora were instrumented with pressure taps midshaft and in the distal diaphysis, a load cell distally to measure force, and a displacement transducer to monitor reamer position. Following initial hand reaming, intramedullary power reaming was conducted utilizing a 9-mm reamer initially, with subsequent increases in steps of 0.5 mm. All femora were maintained at 37 degrees C and albumin was used to maintain a fluid-filled canal. The highest pressures consistently occurred during initial power reaming, with peak pressures ranging from 270 to 1500 mmHg amongst femora with the AO system. No significant differences were found in the peak pressures generated for the two systems (P = 0.10). The pressure measurements at the two locations in the femur were consistently similar, indicating that pressures are continuous throughout this aspect of the femur. The pressures were not correlated with instantaneous applied axial thrust (R2 = 0.191), and this could be attributed chiefly to the additional friction force of cutting. While pressure generation in the medullary canal upon reaming is likely governed by the rate of clearance of canal content, this is a highly variable response produced by characteristics of the femur which are still not fully understood. PMID- 7493782 TI - Improvements in trauma survival in Leeds. AB - One hundred and eighty-six cases of major trauma (Injury Severity Score > 15) admitted to the two Leeds hospitals in 1988/89 were compared with 198 cases admitted in 1992/93 to ascertain if a system of trauma care which had evolved in the hospitals during this time had affected outcome of these patients. Injury severity score, age, and sex distribution were similar for the two groups. There were significantly fewer deaths in 1992/93, particularly in those patients with a score less than 41. It would appear that the emphasis, since 1988, on timely and definitive management of patients with major trauma has led to improved survival of the seriously injured in Leeds. PMID- 7493783 TI - Reduction of post-traumatic osteoporosis after external fixation of tibial fractures. AB - Disuse osteoporosis following tibial fractures is associated with pain and prolonged rehabilitation. In a prospective study, 22 patients with stable tibial fractures treated in plaster of Paris were compared with 15 patients in whom unstable tibial fractures were treated by external fixation. All patients were allowed to bear full weight from the day after the injury. The patients treated with plaster of Paris had the less severe fractures but still had a greater number of the higher grades of osteoporosis. We found a significantly lower degree of osteoporosis in the external fixator group. A comparison of the median percentage bearing weight in the two groups of patients showed greater percentage bearing weight in the external fixator group at four and eight weeks after the injury. We suggest that the reduction of post-traumatic osteoporosis is due to improved early function and weight bearing in patients treated by external fixation. PMID- 7493784 TI - Factors related to the occurrence of postoperative complications following penetrating gastric injuries. AB - The aim of this study was to determine the relationship between diaphragmatic injury and gross contamination of the peritoneal cavity caused by gastric injuries and the occurrence of postoperative complications, especially those related to the pleural cavity. Charts of 73 patients sustaining gastric injuries due to penetrating trauma were retrospectively reviewed. There were 66 males and mean age was 28 years. Stab wounds were the most frequent mechanism of injury, occurring in 46 cases. Most of the injuries were treated using simple suture and minor debridement. Postoperative morbidity rate was 30 per cent and thoracic complications occurred in 11 patients. Twenty-six patients had diaphragmatic injuries; 54 per cent of them developed postoperative complications. Of the remaining 47 patients without diaphragmatic injuries, only eight developed complications. Of the 26 patients with diaphragmatic injuries, seven developed pleuropulmonary complications compared with 4 of 47 without diaphragmatic injury. Of sixteen patients who had gross contamination secondary to gastric injury, characterized by the presence of food or great amounts of gastric contents in the peritoneal cavity, 10 developed postoperative complications compared with 12 of 57 without gross contamination. Overall mortality rate was 11 per cent mostly due to sepsis. In conclusion, the presence of a diaphragmatic injury as well as gross contamination of the abdominal cavity are important factors related to the development of postoperative infections particularly in the pleural space. PMID- 7493786 TI - Assessment of physician competence in prehospital trauma care. AB - In an attempt to develop a model to measure the competence of physicians providing emergency care under difficult field conditions, 75 Israeli army medical corps physicians were evaluated through the use of four instruments: a debriefing interview, peer assessment, self-assessment and written examination. The special on-site assessment model was designed to examine actual events, enabling an assessment of performance in real situations rather than simulated cases. Significant positive correlations were found between the results of the written examination and the peer evaluation on two of four measures (r = 0.36, P = 0.001; r = 0.23, P = 0.05) as well as on the two measures regarding self evaluation and peer evaluation (r = 0.54, P = 0.001; r = 0.38, P = 0.05). It was found that those physicians who were trained in the army's medical officer course scored significantly higher on the written examination (P = 0.001) and were rated more highly by their senior peers (P = 0.048) than those who did not receive such training. It was concluded that it is advantageous to use a combination of knowledge (written examination) and performance (peer assessment or self assessment) measures in order to arrive at a more comprehensive assessment of competence. In addition, the written examination format should be expanded and developed to include more clinical vignettes requiring treatment decisions, making this instrument a more clinically oriented measure of physician competence in trauma care. PMID- 7493785 TI - Craniofacial injuries in unhelmeted riders of motorbikes. AB - We studied 1160 consecutive craniofacial injuries sustained by unhelmeted motorbike riders in Taipei, Taiwan, between 1990 and 1993, in order to investigate the distribution, type and severity of these injuries. The average age of the victims was 31 years (SD 13.2), with 84 per cent of them being between ages 16 and 45. The facial and cranial areas were defined as being separated by the line between eyebrows and ears. The incidence of facial injuries was the same as that of cranial injuries (both 68 per cent). While facial injuries occurred most often in the cheek and chin, most cranial injuries occurred in the forehead and parietal region. Although the majority of facial injuries resulted in mild brain injuries, they may also cause serious cosmetic problems, and some were associated with serious brain damage. Motorbike riders need good face protection. Since most motorbikes in Taipei travel relatively slowly, these results may also apply to bicyclists; in other words, cyclists may also need good face protection. PMID- 7493787 TI - Treatment of traumatic effusion in the elbow joint: a prospective, randomized study of 62 consecutive patients. AB - Sixty-two patients with post-traumatic radiologically visualized effusion in the elbow joint, apparently without bony damage, were randomized to either 1 week of immobilization in a plaster or immediately instructed in active exercises without any immobilization. They were reexamined by an orthopaedic surgeon weekly until recovery. New radiographs were taken after 1 week. Reevaluation of the radiographs by a radiologist revealed seven 'false-positive' effusions, i.e. neither effusion nor fracture, and 21 missed fractures. All but five missed fractures, continued in the study. Thirty patients started immediate active exercises instructed by the surgeon and 27 were immobilized. The 'active exercise' group had a significantly shorter recovery time (one week vs two weeks, P < 0.05). The presence of missed fractures did not influence the result and all patients recovered fully. Also four of the excluded patients with missed fractures recovered fully. One patient with missed fracture dropped out from follow-up. We recommend that an apparently isolated post-traumatic effusion in the elbow joint is treated with immediate active exercises followed by a clinical reexamination after one week supplemented with new radiographs if there is unsatisfactory clinical progress. PMID- 7493788 TI - The suspected scaphoid fracture and isotope bone imaging. AB - Carpal trauma often results in scaphoid injury. A small group of patients have clinical evidence of scaphoid fracture but radiographs fail to demonstrate bony damage. Persistent pain warrants isotope bone scanning. This report reviews the results of 36 consecutive bone scans performed for the suspected scaphoid injury and reviews radiographs taken on presentation. In all, 18 diagnosed fractures: there were three of the scaphoid, six of the distal radius and nine of other carpal bones. It was possible, with hindsight, to see eight fractures on the original radiographs. In patients with continued pain over the radial side of the carpus, attention should be given to the other carpal bones and the distal radius and not just the scaphoid. PMID- 7493789 TI - Stability of reamed and unreamed intramedullary tibial nails: a biomechanical study. AB - We examined the correlation of bone mineral density and bone strength in the cadaver tibia, and looked in vitro at the relative stability of tibial fractures fixed with either reamed or unreamed tibial nails. Bone-mineral density correlated well with bone strength (r = 0.946), but paired tibias did not correlate closely. The unreamed nail-bone construct was less stable than the reamed construct in each pair tested (P < 0.05), and a comparison of all bones showed it to be less stable at all levels of our testing regimen, including failure (P < 0.01). Bending and breakage was seen in four of the smaller unreamed interlocking screws (4 mm). PMID- 7493790 TI - Supracondylar fractures of the distal humerus in children. AB - An audit of 32 displaced supracondylar fractures of the humerus in children treated at the Glasgow Royal Infirmary between June 1990 and September 1992 was carried out. Six fractures were classified as Grade 2 (one cortex intact) and 26 were Grade 3 (no cortical contact). All grade 2 fractures were treated non operatively with good results. Relatively poor results were obtained when displaced fractures were treated non-operatively with manipulation and plaster immobilization. Seven patients underwent manipulation and percutaneous pinning but two developed a cubitus varus deformity. Open reduction and internal fixation with two K-wires gave the best results with no deformity in ten patients. We therefore conclude that this is the optimal method of treatment in a hospital which deals with relatively few completely displaced fractures, with the cosmetic appearance of the scar being minimized by a medial approach. PMID- 7493791 TI - Displaced femoral neck stress fracture in a marathon runner. PMID- 7493792 TI - Early rupture of the arterial anastomoses with free flap survival. PMID- 7493793 TI - Dorsal dislocation of the triquetrum: a rare complication of perilunate dislocation. PMID- 7493794 TI - Groin lymphocele following blunt trauma. PMID- 7493795 TI - Radial head dislocation with radial shaft fracture. PMID- 7493796 TI - 1995 Distinguished Scientific Achievement Award. Presented to Keith Eckerman. PMID- 7493797 TI - 1995 Distinguished Scientific Achievement Award. Presented to Robert G. Thomas. PMID- 7493798 TI - 1995 Distinguished Scientific Achievement Award Memorialization. Presented in memory of Ralph G. Fairchild. PMID- 7493799 TI - 1995 Elda E. Anderson Award. Presented to Kathryn A. Higley. PMID- 7493800 TI - 1995 William McAdams Outstanding Service Award. Presented to William R. Casey. PMID- 7493801 TI - 1995 National Registry of Radiation Protection Technologists Arthur F. Humm, Jr., Memorial Award. Presented to Don W. Marshall. PMID- 7493802 TI - 1995 National Registry of Radiation Protection Technologists Arthur F. Humm, Jr., Memorial Award. Presented to Francis J. Haughey. PMID- 7493803 TI - A limited bibliography of the Federal Government-funded human radiation experiments. AB - From the early 1940's thousands of U.S. citizens have been the subjects of federally supported scientific experiments that involved the administration of ionizing radiation or radioactive substances. Recently, many questions have been raised regarding the nature, scientific value, and ethics of these experiments. Although the results of many of the early human experiments involving radiation have been crucial to the establishment of nuclear medicine, radiation therapy, and radiological protection standards, the underlying ethical basis for a small number of these studies is being questioned. A thorough analysis of these studies and their ethical basis is beyond the scope of this article. Rather, in order to quickly provide the health physics community with some of the available resources in the open literature, a list of bibliographic citations of the 47 studies primarily funded by the Atomic Energy Commission (AEC) and other predecessors of the Department of Energy is presented and briefly summarized. A classification scheme for the human radiation experiments is also developed. PMID- 7493804 TI - A method for computing the decision level for samples containing radioactivity in the presence of background. AB - It is often necessary to make a decision whether or not a sample contains radioactivity in excess of background. The most common procedure is a deterministic approach that compares the result from each sample against a critical level, Lc. Originally Lc was derived from the case where each sample is paired with a blank or control that is known not to contain excess radioactivity. However, most analytic laboratories do not process a blank with each and every sample. Another approach computes Lc using a collection of measurements to form a "well defined" background. This paper presents a method for determining a decision level, L delta, that includes the uncertainties in both the mean and variance of background and how these combine to form uncertainties in estimating the tails of the background distribution. The values of L delta are greater than Lc for a given Type I error. It can be applied whenever the true distribution of background is either normal or log normal. The process is not restricted to counting statistics and is valid for identifying excess contamination of any type providing that unknown samples are processed identically to background samples. PMID- 7493805 TI - Probabilistic risk assessment of nephrotoxic effect of uranium in drinking water. AB - Assessments of the health effects of pollutants generally use models in which exposure variables and model parameters are point values, often chosen as conservative estimates. A more realistic approach is to characterize the uncertainty of each variable and parameter explicitly as a probability distribution. This paper presents a probabilistic version of an established model for metabolism of ingested uranium and characterizes the parameters of the model as distributions. It then characterizes the uncertainty of the threshold level for nephrotoxic risk of uranium, and uses distributions of the heterogeneity in drinking water intake of uranium in the U.S. to assess nephrotoxic risk. It further evaluates the implications of inadequate data on the heterogeneity of dietary intake. PMID- 7493807 TI - Estimated maximum thyroid doses from 129I releases from the Hanford site for the years 1944-1995. AB - 129I, a very long lived fission product, persisted in the environment after discharge from the chemical reprocessing plants at Hanford. The significance to human health is evaluated by considering the maximum possible impact it might have. Allowing the total production of 129I to be discharged to the air and considering the location where the maximally exposed individuals residue, the lifetime cumulative thyroid dose is calculated as a function of birth year. The maximally exposed individual at the maximum exposure location was born in 1954 and accumulates a thyroid dose between 1954 and 1995 of slightly over 9,000 microSv (900 mrem) at an average rate of 220 microSv (22 mrem) y-1. Over the same period of time, the soft tissue dose from background radiation due to both external and internal radiation is about 38,000 microSv (3,800 mRem). PMID- 7493806 TI - A generic model for estimating the risk of deterministic effects of partial organ irradiation by hot particles. AB - A generic (Weibull-type) model is presented for evaluating the risk of a specified deterministic effect of hot-particle (i.e., highly radioactive particle) exposure of the skin, eye, ear, respiratory tract, or gastrointestinal tract. The generic model is a composite of the two-parameter Weibull function used for evaluating the risk of deterministic effects of total-organ irradiation and an isoeffect equation for partial-volume (i.e., fractional-volume) irradiation. Isoeffect refers to a constant level of risk rather than a constant level of severity. The generic model provides a way to evaluate the uniform exposure, isoeffect equivalent dose, H'T,V, to a partial volume V that incurs the same risk as is associated with a nonuniform exposure of volumes within V. When V = 1, the volume is called a reference volume. The reference volume can be associated with a specific organ or mass of tissue. The reference-volume specific, uniform-exposure, isoeffect equivalent dose, H'T,1, can be used to limit the risk of deterministic effects of hot-particle exposure by requiring that H'T,1 remain below the equivalent dose limit for any reference volume of tissue at risk. Local doses H'T,V, can be converted to isoeffect doses H'T,1 by multiplying H'T,V by a volume weighting factor Wv, where Wv equals Vn. For deterministic effects, n < 1. For stochastic effects, n = 1. For stochastic effects, H'T,1 can be converted to an effective dose by multiplying H'T,1 by an appropriate tissue-weighting factor WT. Results presented apply to circumstances where, at most, only a few hot particles are involved. In such circumstances, it is highly unlikely that local tissue will be irradiated by more than one hot particle. Although the focus of this paper is on hot-particle effects, results presented also apply to partial-organ or partial-tissue irradiation by any radiation source including external beams (e.g., electrons, neutrons, protons, heavy ions, and X-ray and gamma-ray photons). PMID- 7493808 TI - Comprehensive appraisal of 239 + 240Pu in soils around Rocky Flats, Colorado. AB - Plutonium contamination of soils around Rocky Flats Environmental & Technology Site, near Golden, Colorado, resulted from past outdoor storage practices and subsequent remobilization due to inadequate cleanup practices. Until now human health risk assessment has not been performed because of a lack of sufficient information regarding the spatial extent of 239 + 240Pu in soils. The purpose of this work was to elucidate the extent of plutonium contamination in surface soils, and to assess the uncertainty associated with the spatial distribution of 239 + 240Pu around Rocky Flats Environmental & Technology Site. Four data sets were collected or compiled for this investigation: (1) samples collected from 240 plots of 1.01- or 4.05-hectare by compositing 25 evenly-spaced samples from the upper 0.64 cm in each plot; (2) samples collected from the upper 5 cm of soil in 167 of the same 240 plots by compositing 10 samples from the center of each plot; (3) historical data compiled from samples collected between 1969 and 1973, considered to be the most indicative of the original release; and (4) the exhaustive data set that contains the samples from 1, 2, and 3 and other published data sets collected between 1974 and 1994. These latter samples varied in depth and method of sampling. Plutonium activity reported in the exhaustive data set ranged from 0.03 Bq kg-1 to 407,000 Bq kg-1 with a mean of 1,443 Bq kg 1, median of 6.6 Bq kg-1, standard deviation of 18,463 Bq kg-1, and a coefficient of variation of 12.6. The technique of nonparametric indicator kriging was used to model four conditional cumulative distribution functions of 239 + 240Pu in soils around Rocky Flats Environmental & Technology Site. Each of the conditional cumulative distribution functions was used to generate an E-type (mean of the conditional cumulative distribution functions) surface. The resulted surfaces were consistent with the hypothesis that the westerly winds were the dominant mechanism of plutonium dispersal. Other processes, such as downstream transport of sediment along local southeast-trending drainages, may have additionally moved small amounts of plutonium. The conditional cumulative distribution functions were also used to construct probability of exceedance maps of 239 + 240Pu in soils. For the purpose of this report two threshold values for the probability maps were selected: (1) the mean and the highest measured global fallout of plutonium (1.48 and 2.96 Bq kg-1, respectively); and (2) the programmatic preliminary remediation goal for residential occupancy scenario (126 Bq kg 1).(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7493809 TI - Measurements of 220Rn emanation from rocks. AB - Alpha-particle and gamma-ray measurements are employed to determine the individual emanation and production rates associated with the thorium radioactive series for thirty-seven rock specimens of approximately 200 cm3 individual volume representing igneous, sedimentary, and metamorphic types. These results are combined to establish 220Rn emanating powers for each of these specimens, and a wide range of values is observed. The generally large 220Rn emanation observed for these specimens strongly indicates that non-tortuous diffusion paths are commonly present in the associated structures, which gives more evidence for the existence of well-connected pore networks at the nanometer level in many of the specimens studied. While the results are qualitatively explainable, they are not predictable in terms of the current macroscopic observables. Since 220Rn and 222Rn emanation rates from rock specimens have previously been found to be comparable in magnitude, the relatively fast determination of 220Rn emanation rates described herein (measurements involving 1-h duration) is in reasonable probability indicative of a comparable 222Rn emanation rate: The employment of 220Rn as a convenient screening tool for potentially high 222Rn emanation sources is indicated. PMID- 7493810 TI - A study of amplifying the response of an LR115 solid state track detector by combining it with electret. AB - Radon progeny are positively charged immediately after formation. A negatively charged electret collects radon progeny atoms which are produced in the diffusion chamber. The detector sensitivity may be increased by using an electret in front of solid state nuclear track detector. Dependence of detection sensitivity on distance between electret and detector LR115 II is studied theoretically and experimentally in this paper. A relatively small fraction of 218Po atoms that formed in the diffusion chamber are collected by the electret. We estimated that the attracted fraction of 218Po was 17% while the attracted fraction of 214Bi 214Po is considerably larger and amounted to approximately 60%. These results confirm previous finding that 218Po atoms discharge quickly after their formation. The comparative radon measurements using diffusion chambers with and without electrets were performed. The amplification of detector sensitivity due to the electret amounted to approximately 80%. PMID- 7493811 TI - Radiocesium distribution in undisturbed soil: measurements and diffusion advection model. AB - The profile of 137Cs, due to Chernobyl accident, in undisturbed soil was measured experimentally over the years 1987 to 1994 and was found to remain practically fixed in the upper 30 cm of soil since 1987. Total deposition of 137Cs at the site was 20 kBq m-2, and approximately 80% of that is in the upper 10 cm. The profile has two slopes in semilogarythmic scale, i.e., it appears as the sum of two exponentials. The contribution of weapons fallout is found to be negligible. Pure diffusion and diffusion-advection models of cesium migration are investigated. It is found that the pure diffusion model cannot reproduce the double slope, while the diffusion advection model can. PMID- 7493812 TI - High-energy gamma rays in Hiroshima and Nagasaki: implications for risk and WR. AB - Based on the DS86 dosimetry system, nearly all of the dose to survivors of the atomic bombings of Hiroshima and Nagasaki was due to unusually high-energy gamma rays, predominantly in the 2- to 5-MeV range. These high energies resulted in part from neutron capture gamma rays as the bomb neutrons penetrated large distances of air. Because of the inverse relationship between energy and biological effectiveness, these high-energy gamma rays are expected to be substantially less effective in producing biological damage than the radiations commonly used in radiobiology and risk assessment. This observation has implications for radiation protection and risk assessment. PMID- 7493813 TI - Retention and excretion of 95Zr-95Nb in humans. AB - This note describes the retention and excretion of 95Zr-95Nb in humans based on a recent CANDU experience and a literature survey of reported cases. Two data bases, QUEST and INIS were used for the survey. Three reported cases were discovered: two for occupational exposures and one for public exposure from nuclear weapons fallout. Human lung retention from these three cases, plus whole body retention and some limited fecal excretion data from a recently occurred exposure at a CANDU station, were reviewed and tested against predictions based on ICRP Publication 30 model. Based on the fits of this model to the reported data it seems that the three occupational exposures exhibit class Y behavior while the public exposure exhibits class W behavior. For only one case is the chemical compound known with certainty: ZrO2. Zirconium oxides are currently classified as class W in ICRP Publications 30 and 54. This work confirms a suggestion that oxides of zirconium be classified as class Y and should be taken into account by the ICRP in its future publications. PMID- 7493814 TI - Study of the occurrence of 222Rn and 226Ra in drinking water in Spain. AB - With the aim of determining the contribution of water to the background of natural radiation in Spain, the 222Rn concentration levels were measured in domestic drinking water from a large part of the country. The water analyzed, both surface and groundwater, was collected in two areas of very distinct geological characteristics, pre-selected for the likelihood of their containing high levels of 222Rn. This water was measured using a coaxial detector of high purity Ge. The overall results demonstrate a log-normal distribution of 222Rn levels, with a geometric and arithmetic means of 10 and 381 Bq L-1, respectively, and a range between detection limit of 2 Bq L-1 and a maximum of 31,000 Bq L-1. The 222Rn concentrations measured in surface water are generally below the detection limit. Significant differences have been found depending on the geological characteristics of the area of reference, although the very distinct concentration levels of 222Rn found in samples of similar lithology imply the influence of more complex factors in the solution of 222Rn in water. Due to the relationship between radon and 226Ra, the concentration of the latter was measured in the same water using radiochemical separation and a scintillation ZnS(Ag) detector. No significant correlation between 226Ra and 222Rn concentrations has been observed unless measurements carried out in a high background radiation region are considered separately. PMID- 7493815 TI - Practical considerations for TLD-400/700-based gamma ray dosimetry for BNCT applications in a high thermal neutron fluence. AB - Operating experience with thermoluminescent dosimeters used in a boron neutron capture therapy research project is reported. In particular, certain facets of the use of thermoluminescent dosimeters for gamma ray dose measurements in the presence of a high thermal neutron fluence are discussed, including a comparison of TLD-400 and TLD-700 for gamma ray dosimetry, annealing procedures, and the effects of neutrons (56Mn activation) on TLD-400. The TLD-400 were observed to have a thermal neutron sensitivity (due to 56Mn beta decay) of 1.5 x 10(-13) Gy per n cm-2. An algorithm was developed to correct for the 56Mn beta decay thermal neutron-induced effects on TLD-400 by using a two-stage thermoluminescent readout for the thermoluminescent dosimeter chips. PMID- 7493816 TI - Workload and use factor of medical linear accelerators in radiotherapy. AB - An important factor in the design of primary protective barriers is the use factor. The present study was aimed at obtaining historical data on the use factor of two dual modality linear accelerators in a radiotherapy department. Gantry angle, field size, and beam modifiers were recorded for all radiation qualities in use at two medical linear accelerators with 6 MV and 18 MV x-rays and multiple electron energies ranging from 4 MeV to 20 MeV. The data for one year of clinical use was extracted from a record and verifying system and an estimate of the physics workload on the machines was obtained by going through the quality assurance records and machine log books. Of the total dose of approximately 37,000 Gy delivered in one year at isocenter on each unit 80% was given as 6 MV x-rays. As can be expected, most x-ray beams were directed at the four cardinal gantry angles with the angular distribution for 6 MV and 18 MV x rays being very similar. Electron fields were broadly distributed around the gantry pointing down position. Less than 25% of all clinical x-ray treatment fields extended beyond a field size of 200 cm2. PMID- 7493817 TI - Radiation protection design philosophy for a door interlock system for shared room remote afterloading brachytherapy. AB - A single remote afterloading system can sometimes be used for the radiation treatment of two or more patients in separate rooms simultaneously. This configuration poses certain radiation protection problems, especially in a busy clinic where some of the treatment rooms have to be used for other non-radiation related patients even though not all radiation treatments have been completed. In this report we describe a door interlock system that has been designed to allow for radiation protection purposes during radiation treatment but is disabled when the radiation treatment is completed--with enough safeguard built in to prevent accidental bypass of the interlock. In addition, the quality control procedures of the radiation monitor devices for these treatment rooms are described. These radiation protection procedures could be generalized to other remote afterloading systems. PMID- 7493818 TI - The national dose registry for radiation workers in Switzerland. AB - The national dose registry began operation in Switzerland in 1990. It is a database maintained by the Federal Office of Public Health. It contains personal, employment, and dosimetric data for all the radiation workers in the country. The data for about 60,000 currently employed radiation workers are transferred from ten external dosimetry services to the registry every month or every three months. The registry was initiated to assist national authorities in controlling and safekeeping of the occupational doses and to allow statistical evaluations. In autumn 1994 the new Radiation Protection Regulations came into effect. Based on the ICRP recommendations, the Regulations introduced lower limits for the annual dose, for the average dose over five years, and for pregnant women and defined the internal dose as an additive component to the total dose. All these items require a high quality registration of occupational doses. This can hardly be achieved without a central registry. This paper describes the design of the Swiss national dose registry in the frame of the particular organization of the personal dosimetry in this country. The database structure and the information content are presented. Special solutions for the data transfer have been found, as the registry is running completely decoupled from the dosimetry services. Applications of the registry for regulatory control, particularly in connection with the new Regulations are described. Some results of statistical analyses are presented. The possibilities and the limitations of using the registry for radioepidemiological studies are discussed. PMID- 7493819 TI - Further response to Watson. PMID- 7493820 TI - Response to Kramer, Hoshi, and Okajima. PMID- 7493821 TI - Ocular surface area as an informative index of visual ergonomics. AB - A large ocular surface area (OSA) is thought to be one of the causes of eye irritation and eye fatigue. Because ocular surface is very sensitive to various irritants such as dust, heat, dryness, air flow, etc., a large OSA increases the possibility of eye surface exposure to such irritants. Thus, OSA is one of the most important indices of visual ergonomics. This paper aims at making OSA an exact and practical index by first describing an accurate method of measuring it, and then clarifying the relationship between OSA, the width of the palpebral fissure, and vertical gaze direction, all of which are thought to be strongly correlated with each other. We derived the following equations: 1) y = 0.039x1 + 3.36, r = 0.99, 2) y = 3.05x2 - 0.39, r = 0.97, 3) x1 = 72.7x2 - 91.4, r = 0.97, where x1 = vertical gaze direction (degrees), x2 = width or the palpebral fissure (cm), y = OSA (cm2). Finally, this paper also introduces the practical applications of OSA measurement, and notes the differences between OSA when VDT work (word processing using a keyboard and drawing a picture using a mouse) is performed and when traditional office work without a VDT (reading, drawing, and writing) is performed. PMID- 7493822 TI - Circadian rhythm of catecholamine excretion in rats after phase shift of light dark cycle. AB - To clarify the time course of circadian rhythm adaptation to a phase shift of the light-dark (LD) cycle, urinary excretion of catecholamines was measured in rats before and after a 12-hour or 6-hour phase delay of a 12-hour light and 12-hour dark schedule. In rats under a basal condition, distinct circadian rhythms in catecholamine excretion were observed, especially in adrenaline excretion. During the 1st and 2nd days after a 12-hour phase delay, the acrophase and amplitude of adrenaline rhythm remained almost unchanged, but thereafter the acrophase was retarded and the amplitude was reduced. The acrophase once again became constant after 5 or 6 days, but the ratio of amplitude to mesor in the circadian rhythm of adrenaline excretion and the ratio of light-period to 24-hour noradrenaline excretion were readjusted to the new LD schedule on 11th or 12th day. IN the 6 hour phase delay of the LD cycle, similar findings were observed, and the results suggested adaptation on the 5-6th day. It is considered that the circadian rhythms of the sympathetic adrenomedullary function are restored, at the latest, 12 days after a 12-hour delay of the LD cycle, and 6 days after a 6-hour delay, suggesting that rats need approximately 1 day to adapt to a 1-hour phase shift. PMID- 7493823 TI - A prospective study on mortality among Japanese coal miners. AB - We conducted a prospective study to clarify mortality patterns among Japanese coal miners in a former coal mining area. Subjects included 1,796 coal miners and 4,022 non-coal-miners, who were identified by a mail survey between 1987 and 1989, and then followed up from the date of the survey to April 30th, 1994. We applied Cox's proportional hazards model to compare the mortalities between coal miners and non-coal-miners. Among the coal miners, significantly high risk ratios were observed in all causes of death (risk ratio = 1.4, p < 0.05) and all malignant neoplasms (risk ratio = 1.5, p < 0.05). Risk ratios for all causes of death and all malignant neoplasms also rose with the length of experience in coal mining. Analysis of the results for sites of cancer showed that coal miners had high risk ratios for stomach cancer (risk ratio = 1.6), liver cancer (risk ratio = 1.4) and lung cancer (risk ratio = 1.6), though these ratios were not statistically significant. When the risk ratio for lung cancer was analyzed according to the length of experience in coal mining, coal miners with at least 15 years' experience had a significantly high risk ratio (risk ratio = 2.4, p < 0.05), though coal miners with less than 15 years' experience had almost the same risk as non-coal-miners. PMID- 7493824 TI - Effects of marital status and position on personality traits in engineers of a computer manufacturing plant. AB - To investigate the effects of marital status and position on DSM-III-R personality traits, we administered the self-rating questionnaires of the Structured Clinical Interview for DSM-III-R Personality Disorders to 757 male computer engineers, aged 20 to 49 years, at a computer manufacturing plant. The scores for six of eleven personality traits were significantly higher in single computer engineers than in married ones. Also, among the engineers aged 40 to 49, the scores for six personality traits of section-heads were significantly higher than those among persons in the other positions. These findings suggest that "maladaptive" personality traits, defined by the DSM-III-R, differ with marital status and position in a company. PMID- 7493825 TI - Efficacy of liposome encapsulated triethylenetetraamine hexaacetic acid (TTHA) against cadmium intoxication: role of lipid composition. AB - Efficacy of Triethylenetetraamine hexaacetic acid (TTHA) encapsulated in liposomes having different lipid compositions was examined in animals pre-exposed to cadmium. Mice were injected with cadmium as cadmium (II) chloride 0.5 mg/kg b. wt. intraperitoneally daily for five days. Four weeks after the last injection of cadmium they were administered three injections of TTHA encapsulated in liposomes composed of either phosphatidyl choline:cholesterol (PC:Chol) or sphingomyelin:cholesterol (SM:Chol) in 1:1 molar ratio at a gap of 48 h. Urinary and fecal elimination of cadmium and its distribution in liver, kidneys and spleen were examined. Treatment with TTHA encapsulated in liposomes mobilized higher amount of cadmium from liver and spleen. The overall efficiency for cadmium mobilization was better in TTHA encapsulated in SM:Chol liposome treated group which also led to enhanced excretion of cadmium through urine and feces. The results indicate that TTHA encapsulated in SM:Chol liposomes exhibited highest efficacy in mobilizing cadmium from the body of pre-exposed mice followed by PC:Chol liposomes and the free drug. PMID- 7493826 TI - The treatment of stage III nonsmall cell lung cancer using high dose conformal radiotherapy. AB - PURPOSE: To review our experience using conformal treatment planning and high dose radiotherapy for Stage IIIa and IIIb nonsmall cell lung cancer (NSCLC), and to identify a subset of patients best suited for this approach by analyzing multiple pretreatment patient and tumor characteristics. METHODS AND MATERIALS: Between December 1987 and June 1992, 37 patients with Stage III NSCLC treated with high-dose radiotherapy using conformal radiotherapy were reviewed. The patient characteristics were as follows: Stage IIIa (18 patients), IIIb [19]; T1 2 [13], T3-4 [24]; N0-1 [8], N2-3 [29]; and median age 63. All patients were treated with 1.8-2.0 Gy fractions to a median dose of 66 Gy (range 60-70 Gy). Outcome was analyzed by multiple pretreatment variables including age, sex, Karnofsky performance score, pretreatment symptoms, stage group, T and N stage, tumor volume (calculated from computed tomography (CT) contours), presence of atelectasis, and tumor histology. Outcome was also analyzed by total radiotherapy dose. RESULTS: The median, 1-year and 2-year survival rates for the entire group were 19.5 months, 75 and 37%, respectively. The median, 1-year, and 2-year local progression-free survival rates are 15.6 months, 62 and 23%. There was no difference in survival by stage group (IIIa vs. IIIb) or by T or N stage. Tumor volumes ranged from 47-511 cc in the patients without atelectasis and were not a significant prognostic factor. Histology was found to be a significant prognostic factor, with squamous cell carcinoma having a better overall survival and local progression-free survival than other histologies. No other patient characteristic was found to be significant by either univariate or multivariate analysis. When outcome was analyzed by radiotherapy dose, no dose response was evident in the narrow dose range studied (60-70 Gy). Toxicity included two cases of pneumonitis, which resolved with conservative therapy. CONCLUSION: High-dose conformal radiotherapy, in our experience, results in overall survival rates that compare favorably with trials of chemoradiotherapy or conventional radiotherapy with a low treatment-associated morbidity. However, local progression remains a significant problem despite median radiotherapy doses of 66 Gy. Future trials using escalating radiotherapy doses with conformal radiotherapy are therefore, indicated. PMID- 7493827 TI - The potential for normal tissue dose reduction with neoadjuvant hormonal therapy in conformal treatment planning for stage C prostate cancer. AB - PURPOSE: Preirradiation hormonal cytoreduction of prostate cancer has been proven to reduce exposure of normal structures by decreasing the size of the target volume. Dose-volume histogram (DVH) analysis, however, does not always appear to demonstrate a strong positive benefit with the use of neoadjuvant hormone therapy. This study analyzes various other factors influencing dose to normal organs, which may determine the success or failure of neoadjuvant hormonal therapy in achieving its goals. METHODS AND MATERIALS: Patients with bulky clinical Stage C adenocarcinoma of the prostate were given 3 months of hormone treatment consisting of oral Flutamide and monthly Zoladex injections prior to irradiation. Computerized tomography (CT) scans of the pelvis were obtained both prior to and following hormonal treatment. Treatment plans were generated by three-dimensional (3D) conformal treatment planning. The change in the volume of the prostate was assessed along with the percentage of prescribed dose delivered to the rectum and bladder. Various factors such as prostate size, bladder/rectum size, and organ shape were studied. Both dose-volume histograms (DVH) and dose surface area histograms (DSH) were used for analysis. RESULTS: Six of seven patients had reduction in the size of their prostates. The mean volumes of the prostate before and after hormonal manipulation were 129.1 +/- 32.9 standard deviation (SD) cm3 and 73.0 +/- 29.5 SD cm3, respectively (p = 0.0059). The volume of rectum receiving 80% of the prescribed dose was reduced in five of seven patients from a mean of 83.2 to 59.9 cm3 (p = 0.045). The volume of bladder receiving 80% of the prescribed dose was also reduced in five out of seven patients from a mean of 74.5 to 40.2 cm3 (p = 0.098). Correlation between the size of the prostate and volume of rectum and bladder treated was not always consistent: greater reduction in prostate size did not necessarily result in large decreases in dose to bladder or rectum. The total size of the bladder and rectum were found to be important factors in normal tissue radiation exposure; the benefits of hormone therapy may be lost if the bladder and rectum are allowed to decrease in size. Also, the bladder may be prone to sagging into the pelvis of some patients following hormone therapy, resulting in a less optimal therapeutic ratio. CONCLUSION: Reduction in prostate size by neoadjuvant hormonal manipulation does decrease the amount of normal tissue irradiated in most patients. However, the correlation between the reduction in prostate size and amount of rectum or bladder treated is not linear if other variables are not controlled. Factors such as the shape of the organs, as well as the distensible nature of the bladder and rectum, play major roles in dose to normal tissues. These facts may mask the benefits of cytoreduction and could be obstacles in realizing consistent benefits from preirradiation hormonal treatment in the clinical setting if they are ignored. PMID- 7493828 TI - Conformal prostate brachytherapy: initial experience of a phase I/II dose escalating trial. AB - PURPOSE: To improve treatment results on prostatic adenocarcinoma, conformal radiation therapy (CRT) has been used. Two major drawbacks of external CRT are: (a) internal organ motion/daily set-up variations, and (b) exclusion of several patients for CRT based on poor geometrical relationships as identified by three dimensional (3D) treatment planning. To overcome the above problems, we began the first prospective Phase I/II dose-escalating clinical trial of conformal brachytherapy (CB) and concurrent external beam irradiation. METHODS AND MATERIALS: Fifty-nine patients with T2b-T3c prostatic adenocarcinoma received 176 transperineal ultrasound-guided conformal high-dose rate (HDR) boost implants. All patients received concomitant external beam pelvic irradiation. Dose escalation of the three HDR-CB fractions proceeded as follows: 5.5 Gy (30 patients), 6 Gy (20 patients), and 6.5 Gy (9 patients). The CB dose was prescribed to the prostate contour as outlined using an online biplanar transrectal ultrasound probe. The urethra, anterior rectal wall, and prostate boundaries were identified individually and outlined at 5 mm intervals from the base to the apex of the gland. The CB using real-time ultrasound guidance with interactive online isodose distributions was performed on an outpatient basis. As needles were placed into the prostate, corrections for prostate displacement were recorded and the isodose distributions were recalculated to represent the new relationship between the needles, prostate, and normal structures. No computerized tomography (CT) planning or implant preplanning was required. RESULTS: No patient was rejected based on poor geometrical relation of pelvic structures. In every implant performed, prostate displacement was noted. Craniocaudal motion of the gland ranged from 0.5-2.0 cm (mean = 1.0 cm), whereas lateral displacement was 0.1-0.4 cm. With the interactive online planning system, organ motion was immediately detected, accounted for, and corrected prior to each HDR treatment. The rectal dose has ranged from 45 to 87%, and the urethral dose from 97 to 112% of the prostate dose. It is significant to note that operator dependence has been completely removed because the interactive online planning system uniformly guides the physicians. CONCLUSIONS: With ultrasound guidance and the interactive online dosimetry system, organ motion (as compared to external beam) is insignificant because it can be corrected during the procedure without increasing target volume margins. Common pitfalls of brachytherapy, including operator dependence and difficulty with reproducibility, have been eliminated with the intraoperative online planning system. PMID- 7493829 TI - The current state of the art in three dimensional oncologic imaging: an overview. AB - PURPOSE: To provide an overview of the methods and clinical applications of three dimensional (3D) medical imaging in the oncologic patient. METHODS AND MATERIALS: We briefly outline the techniques currently used to create 3D medical images with an emphasis on their strengths and shortcomings as they relate to oncologic imaging and radiation therapy planning. We then discuss some of the most important and promising oncologic applications of 3D imaging and suggest likely future directions in this rapidly developing field. RESULTS: Since the first application of 3D techniques to medical data over a decade ago, 3D medical images have evolved from relatively crude representations of musculoskeletal abnormalities to detailed and accurate representations of a variety of soft tissue, vascular, and oncologic pathology. The rapid development of both computer hardware and software coupled with the application of 3D techniques to a variety of imaging modalities have expanded the clinical applications of this technology dramatically. CONCLUSIONS: 3D medical images are clinically practical tools for oncologic evaluation and effective radiation therapy planning. PMID- 7493830 TI - Tumor and target delineation: current research and future challenges. AB - In the past decade, significant progress has been made in the imaging of tumors, three dimensional (3D) treatment planning, and radiation treatment delivery. At this time one of the greatest challenges for conformal radiation therapy is the accurate delineation of tumor and target volumes. The physician encounters many uncertainties in the process of defining both tumor and target. The sources of these uncertainties are discussed, as well as the issues requiring study to reduce these uncertainties. PMID- 7493831 TI - An image correlation procedure for digitally reconstructed radiographs and electronic portal images. AB - PURPOSE: To study a procedure that uses megavoltage digitally reconstructed radiographs (DRRs) calculated from patient's three-dimensional (3D) computed tomography (CT) data as a reference image for correlation with on-line electronic portal images (EPIs) to detect patient setup errors. METHODS AND MATERIALS: Megavoltage DRRs were generated by ray tracing through a modified volumetric CT data set in which CT numbers were converted into linear attenuation coefficients for the therapeutic beam energy. The DRR transmission image was transformed to the grayscale window of the EPI by a histogram-matching technique. An alternative approach was to calibrate the transmission DRR using a measured response curve of the electronic portal imaging device (EPID). This forces the calculated transmission fluence values to be distributed in the same range as that of the EPID image. A cross-correlation technique was used to determine the degree of alignment of the patient anatomy found in the EPID image relative to the reference DRR. RESULTS: Phantom studies demonstrated that the correlation procedure had a standard deviation of 0.5 mm and 0.5 degrees in aligning translational shifts and in-plane rotations. Systematic errors were found between a reference DRR and a reference EPID image. The automated grayscale image correlation process was completed within 3 s on a workstation computer or 12 s on a PC. CONCLUSION: The alignment procedure allows the direct comparison of a patient's treatment portal designed with a 3D planning computer with a patient's on-line portal image acquired at the treatment unit. The image registration process is automated to the extent that it requires minimal user intervention, and it is fast and accurate enough for on-line clinical applications. PMID- 7493832 TI - Advanced interactive planning techniques for conformal therapy: high level beam descriptions and volumetric mapping techniques. AB - PURPOSE: To aid in design of conformal radiation therapy treatment plans involving many conformally shaped fields, this work investigates the use of two methodologies to enhance the ease of interactive treatment planning: high-level beam constructs and beam's-eye view volumetric mapping. METHODS AND MATERIALS: High-performance computer graphics running on various workstations using a graphical visualization system (AVS) have been used in this work. Software specific to this application has been written in standard FORTRAN and C languages. A new methodology is introduced by defining radiation therapy "fields" to be composed of multiple beam "segments." Fields can then be defined as higher level entities such as arcs, cones, and other shapes. A "segmental cone" field, for example, is defined by a symmetry axis and a cone angle, and can be used to rapidly place a series of beam segments that converge at the target volume, while reducing the degree of overlap elsewhere. A new beam's-eye view (BEV) volumetric mapping technique is presented to aid in selecting the placement of conformal radiation fields. With this technique, the relative average dose within an organ of interest is calculated for a sampling of isocentric, conformally shaped beams and displayed either as a "globe," which can be combined with the display of anatomical surfaces, or as a two-dimensionally mapped projection. The dose maps from multiple organs can be generated, stacked, or composited with relative weightings to aid in the placement of fields that minimize overlap with critical structures. RESULTS: The use of these new methodologies is demonstrated for prostate and lung treatment sites and compared to conventional planning techniques. DISCUSSION: The use of many beams for conformal treatment delivery is difficult with current interactive planning. The use of high-level beam constructs provides a means to quickly specify, place, and configure multiple beam arrangements. The BEV volumetrics aids in the placing of fields, which minimize involvement with critical normal tissues. CONCLUSIONS: Early experience with the new methodologies suggest that the new methods help to enhance (or at least speed up) the ability of a treatment planner to create optimal radiation treatment field arrangements. PMID- 7493833 TI - Three dimensional planning target volumes: a model and a software tool. AB - PURPOSE: Three dimensional (3D) target volumes are an essential component of conformal therapy because the goal is to shape the treatment volume to the target volume. The planning target volume (PTV) is defined by ICRU 50 as the clinical target volume (CTV) plus a margin to ensure that the CTV receives the prescribed dose. The margin must include all interfractional and intrafractional treatment variations. This paper describes a software tool that automatically generates 3D PTVs from CTVs for lung cancers and immobile head and neck cancers. METHODS AND MATERIALS: Values for the interfractional and intrafractional treatment variations were determined by a literature review and by targeted interviews with physicians. The software tool is written in Common LISP and conforms to the specifications for shareable software of the Radiotherapy Treatment Planning Tools Collaborative Working Group. RESULTS: The tool is a rule-based expert system in which the inputs are the CTV contours, critical structure contours, and qualitative information about the specific patient. The output is PTV contours, which are a cylindrical expansion of the CTV. A model for creating PTVs from CTVs is embedded in the tool. The interfractional variation of setup uncertainty and the intrafractional variations of movement of the CTV (e.g., respiration) and patient motion are included in the model. Measured data for the component variations is consistent with modeling the components as independent samples from 3D Gaussian distributions. The components are combined using multivariate normal statistics to yield the cylindrical expansion factors. Rules are used to represent the values of the components for certain patient conditions (e.g., setup uncertainty for a head and neck patient immobilized in a mask). The tool uses a rule interpreter to combine qualitative information about a specific patient with rules representing the value of the components and to enter the appropriate component values for that patient into the cylindrical expansion formula. CONCLUSION: The portable software tool allows the rapid, consistent, and automatic generation of 3D PTVs from CTVs. PMID- 7493834 TI - Three dimensional conformal treatment planning with multileaf collimators. AB - PURPOSE: Three dimensional conformal radiation treatments are complex, often involving large numbers of blocked or multileaf collimated fields that shape regions of high dose to conform to the treatment volume. As manual definition and digitization of aperture shapes and their corresponding multileaf configurations can be impractically time consuming, it was necessary to integrate the planning of multileaf fields into an existing three dimensional treatment planning system and improve the efficiency of treatment delivery to make these treatments feasible on a routine basis. METHODS AND MATERIALS: A subfunction of the Beam's Eye View (BEV) component can be used to automatically generate a continuous aperture shape with a margin around the tumor to account for beam penumbra, and excluding any normal structures to be spared (each with its own margin). To convert a continuous aperture shape into one defined by the multileaf collimator (MLC), a leaf coverage mode is chosen to determine how leaves are fitted to aperture shapes. The conversion process also considers parameters of the specific MLC system, e.g., leaf thickness and the number of leaves. If normal structures to be shielded split the target into multiple regions, more than one multileaf aperture can result. An interactive leaf adjustment routine is also provided to allow for modification of individual leaf positions. Dose calculation programs then take into account multileaf apertures for computation of dose distributions using a pencil beam convolution model. Finally, prescription files specifying leaf and jaw configurations are prepared in treatment machine specific formats and downloaded to the computers driving the multileaf collimators and other components of the treatment machines. RESULTS AND CONCLUSION: An example is presented of a prostate treatment plan, with MLC configurations, dose distributions, and treatment delivery description, along with discussion of clinical implementation at Memorial Hospital. PMID- 7493835 TI - Comparison of simulated annealing algorithms for conformal therapy treatment planning. AB - PURPOSE: The efficiency of four fast simulated annealing algorithms for optimizing conformal radiation therapy treatment plans was studied and the resulting plans were compared with each other and to optimized conventional plans. METHODS AND MATERIALS: Four algorithms were selected on the basis of their reported successes in solving other minimization problems: fast simulated annealing with a Cauchy generating function, fast simulated annealing with a Lorentzian generating function, variable step size generalized simulated annealing (VSGSA), and very fast simulated reannealing (VFSR). They were tested on six clinical cases using a multiple beam coplanar conformal treatment technique. Relative beam weights were computed that maximized the minimum tumor dose subject to dose-volume constraints on normal organ doses. Following some initial tuning of the annealing parameters, each algorithm was applied identically to each test case. Optimization tests were run using different random number sequences and different numbers of iterations. RESULTS: The VSGSA algorithm consistently produced the best results. Using long run times, it generated plans with the highest minimum tumor dose in five of the six cases. For the short run times, the VSGSA solutions averaged larger minimum tumor doses than those of the other algorithms for all six patients, with increases ranging from 0.4 to 5.9 Gy. For three of the patients, the conformal plan gave a clinically significant increase in the minimum tumor dose over the conventional plan, ranging from 8.2 to 13.0 Gy. In two other cases, there was little difference between the two treatment approaches. For one case, the optimized conventional plan was much better than the conformal plan because the conventional beam arrangement included wedges, which offset the multiple beam advantage of the conformal plans. CONCLUSIONS: For equal computing times of both long and short duration, the VSGSA algorithm consistently produced conformal plans that were superior to those produced by the other algorithms. The simple conformal technique used in this study showed a significant potential advantage in the treatment of abdominal tumors. In three of the cases, the conformal plans showed clinically important increases in tumor dose over optimized conventional plans. PMID- 7493836 TI - Collision detection and avoidance during treatment planning. AB - PURPOSE: To develop computer software that assists the planner avoid potential gantry collisions with the patient or patient support assembly during the treatment planning process. METHODS AND MATERIALS: The approach uses a simulation of the therapy room with a scale model of the treatment machine. Because the dimensions of the machine and patient are known, one can calculate a priori whether any desired therapy field is possible or will result in a collision. To assist the planner, we have developed a graphical interface enabling the accurate visualization of each treatment field configuration within a "room's eye view" treatment planning window. This enables the planner to be aware of, and alleviate any potential collision hazards. To circumvent blind spots in the graphic representation, an analytical software module precomputes whether each update of the gantry or turntable position is safe. RESULTS: If a collision is detected, the module alerts the planner and suggests collision evasive actions such as either an extended distance treatment or the gantry angle of closest approach. CONCLUSIONS: The model enables the planner to experiment with unconventional noncoplanar treatment fields, and immediately test their feasibility. PMID- 7493837 TI - Variation in volumes, dose-volume histograms, and estimated normal tissue complication probabilities of rectum and bladder during conformal radiotherapy of T3 prostate cancer. AB - PURPOSE: To determine the pattern of changes of rectum and bladder structures during conformal therapy of T3 prostate cancer and the impact of these changes on the accuracy of the dose-volume histograms (DVHs) and normal tissue complication probabilities (NTCPs) of these organs, based on the planning computed tomography (CT) scan only. METHODS AND MATERIALS: For 11 T3 prostate cancer patients treated with conformal therapy, three repeat CT scans were made in Weeks 2, 4, and 6 of the treatment. The bony anatomy was aligned with the planning CT scan, using three dimensional (3D) chamfer matching. The internal and external surfaces of rectum and bladder were contoured in each scan. Three volumes were calculated for each organ: solid organ (including filling), filling, and wall volume. DVHs and NTCPs were calculated for all structures. RESULTS: The solid organ and filling volumes varied considerably between patients and within a patient and they decreased with increasing treatment time. The largest patient variation was seen for patients with large initial filling volumes. The variations of rectum and bladder wall volumes during treatment were 9 and 17% (1 standard deviation (SD)), respectively, with no time trend. The changes of the high dose (> 80 and 90% of the prescribed dose) volumes of the rectum in response to rectum filling differences were proportional to the whole rectum volume changes. The variation of the high-dose rectum wall volume was relatively small (14%, 1 SD). As a result, the NTCPs of rectum and rectum wall were the same overall and the variation of the NTCPs during treatment was about 14% (1 SD) and not correlated with rectum filling. The variation of the high-dose bladder volumes (about 14%, 1 SD) was smaller than the variation of the whole bladder volumes (30%, 1 SD). The high-dose bladder wall volume decreased significantly due to wall distention as the bladder filling increased. As a result of this complex pattern, the variation of NTCPs of bladder (85%, 1 SD) and bladder wall (88%, 1 SD) during treatment was large and significantly correlated with bladder filling. CONCLUSIONS: The planning CT scan overestimates rectum and bladder filling during treatment. Furthermore, the variation of filling is so large that only the wall structures have relatively constant volumes during treatment. For the rectum wall, the DVHs and NTCPs, as estimated from the initial scan, are representative for the whole treatment, because no correlation was seen between these parameters and organ filling. For the bladder wall, however, such a correlation was present and consequently, the initial bladder wall DVHs and NTCPs can only be representative for the whole treatment, if the bladder filling can be kept reasonably constant during treatment. PMID- 7493839 TI - Computer-aided design and fabrication of an electron bolus for treatment of the paraspinal muscles. AB - PURPOSE: Demonstrate the technology for the design, fabrication, and verification of an electron bolus used in the preoperative irradiation of a mesenchymal chondrosarcoma in the paraspinal muscle region (T8-T12), in which the target volume overlay a portion of the spinal cord, both lungs, and the right kidney. METHODS AND MATERIALS: An electron-bolus design algorithm implemented on a three dimensional (3D) radiotherapy treatment planning system designed the bolus to yield a dose distribution that met physician-specified clinical criteria. Electron doses were calculated using a 3D electron pencil-beam dose algorithm. A computer-driven milling machine fabricated the bolus from modeling wax, machining both the patient surface and the beam surface of the bolus. Verification of the bolus fabrication was achieved by repeating the patient's computed tomography (CT) scan with the fabricated bolus in place (directly on the posterior surface of the prone patient) and then recalculating the patient's dose distribution using the 3D radiotherapy treatment planning system. RESULTS: A treatment plan using a 17-MeV posterior electron field with a bolus delivered a superior dose distribution to the patient than did the same plan without a bolus. The bolus plan delivered a slightly increased dose to the target volume as a result of a slightly broader range of doses. There were significant reductions in dose to critical structures (cord, lungs, and kidney) in the bolus plan, as evidenced by dose-volume histograms (DVHs). The patient dose distribution, calculated using CT scan data with the fabricated bolus, showed no significant differences from the planned dose distribution. CONCLUSIONS: A bolus can provide considerable sparing of normal tissues when using a posterior electron beam to irradiate the paraspinal muscles. Bolus design and fabrication using the tools described in this paper are adequate for patient treatment. CT imaging of the patient with the bolus in place followed by calculation of the patient's dose distribution demonstrated a useful method for verification of the bolus design and fabrication process. PMID- 7493838 TI - A method of analyzing rectal surface area irradiated and rectal complications in prostate conformal radiotherapy. AB - PURPOSE: To develop a method of analyzing rectal surface area irradiated and rectal complications in prostate conformal radiotherapy. METHODS AND MATERIALS: Dose-surface histograms of the rectum, which state the rectal surface area irradiated to any given dose, were calculated for a group of 27 patients treated with a four-field box technique to a total (tumor minimum) dose ranging from 68 to 70 Gy. Occurrences of rectal toxicities as defined by the Radiation Therapy Oncology Group (RTOG) were recorded and examined in terms of dose and rectal surface area irradiated. For a specified end point of rectal complication, the complication probability was analyzed as a function of dose irradiated to a fixed rectal area, and as a function of area receiving a fixed dose. Lyman's model of normal tissue complication probability (NTCP) was used to fit the data. RESULTS: The observed occurrences of rectal complications appear to depend on the rectal surface area irradiated to a given dose level. The patient distribution of each toxicity grade exhibits a maximum as a function of percentage surface area irradiated, and the maximum moves to higher values of percentage surface area as the toxicity grade increases. The dependence of the NTCP for the specified end point on dose and percentage surface area irradiated was fitted to Lyman's NTCP model with a set of parameters. The curvature of the NTCP as a function of the surface area suggests that the rectum is a parallel structured organ. CONCLUSIONS: The described method of analyzing rectal surface area irradiated yields interesting insight into understanding rectal complications in prostate conformal radiotherapy. Application of the method to a larger patient data set has the potential to facilitate the construction of a full dose-surface complication relationship, which would be most useful in guiding clinical practice. PMID- 7493840 TI - A computer-controlled conformal radiotherapy system. I: Overview. AB - PURPOSE: Equipment developed for use with computer-controlled conformal radiotherapy (CCRT) treatment techniques, including multileaf collimators and/or computer-control systems for treatment machines, are now available. The purpose of this work is to develop a system that will allow the safe, efficient, and accurate delivery of CCRT treatments as routine clinical treatments, and permit modifications of the system so that the delivery process can be optimized. METHODS AND MATERIALS: The needs and requirements for a system that can fully support modern computer-controlled treatment machines equipped with multileaf collimators and segmental or dynamic conformal therapy capabilities have been analyzed and evaluated. This analysis has been used to design and then implement a complete approach to the delivery of CCRT treatments. RESULTS: The computer controlled conformal radiotherapy system (CCRS) described here consists of a process for the delivery of CCRT treatments, and a complex software system that implements the treatment process. The CCRS system described here includes systems for plan transfer, treatment delivery planning, sequencing of the actual treatment delivery process, graphical simulation and verification tools, as well as an electronic chart that is an integral part of the system. The CCRS system has been implemented for use with a number of different treatment machines. The system has been used clinically for more than 2 years to perform CCRT treatments for more than 200 patients. CONCLUSIONS: A comprehensive system for the implementation and delivery of computer-controlled conformal radiation therapy (CCRT) plans has been designed and implemented for routine clinical use with multisegment, computer-controlled, multileaf-collimated conformal therapy. The CCRS system has been successfully implemented to perform these complex treatments, and is considered quite important to the clinical use of modern computer-controlled treatment techniques. PMID- 7493841 TI - A computer-controlled conformal radiotherapy system. II: Sequence processor. AB - PURPOSE: A sequence processor (SP) is described as part of a larger computer controlled conformal radiotherapy system (CCRS). The SP provides the means to accept and then translate highly sophisticated radiation therapy treatment plans into vendor specific instructions to control treatment delivery on a computer controlled treatment machine. METHODS AND MATERIALS: The sequence processor (SP) is a small workstation computer that interfaces to the control computer of computer-controlled treatment machines, and to other parts of the larger CCRS system. The system reported here has been interfaced to a computer-controlled racetrack microtron with two treatment gantries, and also to other linear accelerator treatment machines equipped with multileaf collimators. An extensive design process has been used in defining the role of the SP within the context of the larger CCRS project. Flexibility and integration with various components of the project, including databases, treatment planning system, graphical simulator, were key factors in the development. In conjunction with the planned set of treatment fields, a procedural scripting language is used to define the sequence of treatment events that are performed, including operator interactions, communications to other systems such as dosimetry and portal imaging devices, and database management. RESULTS: A flexible system has been developed to allow investigation into procedural steps required for simulating and delivering complex radiation treatments. The system has been used to automate portions of the acceptance testing for the control system of the microtron, and is used for routine daily quality assurance testing. The sequence processor system described here has been used to deliver all clinical treatments performed on the microtron system in 2 years of clinical treatment (more than 200 patients treated to a variety of treatment sites). CONCLUSIONS: The sequence processor system has enabled the delivery of complex treatment using computer-controlled treatment machines. The flexibility of the system allows integration with secondary devices and modification of procedural steps, making it possible to develop effective techniques for insuring safe and efficient computer-controlled conformal radiation therapy treatments. PMID- 7493842 TI - A computer-controlled conformal radiotherapy system. III: Graphical simulation and monitoring of treatment delivery. AB - PURPOSE: Safe and efficient delivery of radiotherapy using computer-controlled machines requires new procedures to design and verify the actual delivery of these treatments. Graphical simulation and monitoring techniques for treatment delivery have been developed for this purpose. METHODS AND MATERIALS: A graphics based simulator of the treatment machine and a set of procedures for creating and manipulating treatment delivery scripts are used to simulate machine motions, detect collisions, and monitor machine positions during treatment. The treatment delivery simulator is composed of four components: a three-dimensional dynamic model of the treatment machine; a motion simulation and collision detection algorithm, user-interface widgets that mimic the treatment machine's control and readout devices; and an icon-based interface for creating and manipulating treatment delivery scripts. These components are used in a stand-alone fashion for interactive treatment delivery planning and integrated with a machine control system for treatment implementation and monitoring. RESULTS: A graphics-based treatment delivery simulator and a set of procedures for planning and monitoring computer-controlled treatment delivery have been developed and implemented as part of a comprehensive computer-controlled conformal radiotherapy system. To date, these techniques have been used to design and help monitor computer controlled treatments on a radiotherapy machine for more than 200 patients. Examples using these techniques for treatment delivery planning and on-line monitoring of machine motions during therapy are described. CONCLUSION: A system that provides interactive graphics-based tools for defining the sequence of machine motions, simulating treatment delivery including collision detection, and presenting the therapists with continual visual feedback from the treatment machine has been successfully implemented for routine clinical use as part of an overall system for computer-controlled conformal radiotherapy treatment, and is considered a necessary part of the routine treatment methodology. PMID- 7493843 TI - A computer-controlled conformal radiotherapy system. IV: Electronic chart. AB - PURPOSE: The design and implementation of a system for electronically tracking relevant plan, prescription, and treatment data for computer-controlled conformal radiation therapy is described. METHODS AND MATERIALS: The electronic charting system is implemented on a computer cluster coupled by high-speed networks to computer-controlled therapy machines. A methodical approach to the specification and design of an integrated solution has been used in developing the system. The electronic chart system is designed to allow identification and access of patient specific data including treatment-planning data, treatment prescription information, and charting of doses. An in-house developed database system is used to provide an integrated approach to the database requirements of the design. A hierarchy of databases is used for both centralization and distribution of the treatment data for specific treatment machines. RESULTS: The basic electronic database system has been implemented and has been in use since July 1993. The system has been used to download and manage treatment data on all patients treated on our first fully computer-controlled treatment machine. To date, electronic dose charting functions have not been fully implemented clinically, requiring the continued use of paper charting for dose tracking. CONCLUSIONS: The routine clinical application of complex computer-controlled conformal treatment procedures requires the management of large quantities of information for describing and tracking treatments. An integrated and comprehensive approach to this problem has led to a full electronic chart for conformal radiation therapy treatments. PMID- 7493844 TI - Clinical implementation of a commercial multileaf collimator: dosimetry, networking, simulation, and quality assurance. AB - PURPOSE: Clinical implementation of multileaf collimation (MLC) includes commissioning (including leaf calibration), dosimetric measurements (penumbra, transmission, calculation parameters), shaping methods, networking for file transfer, verification simulation, and development of a quality assurance (QA) program. Differences of MLC and alloy shaping in terms of penumbra and stair-step effects must be analyzed. METHODS AND MATERIALS: Leaf positions are calibrated to light field. The resultant decrement line, penumbras, leaf transmission data, and isodoses in various planes were measured with film. Penumbra was measured for straight edges and corners, in various media. Ion chambers were used to measure effects of MLC on output, scatter, and depth dose. We maintain midleaf intersection criteria. MLC fields are set 7 mm beyond planning target volumes. After shaping by vendor software or by our three-dimensional planning system, files are transferred to the MLC workstation by means of sharing software, interface cards, and cabling. A MLC emulator was constructed for simulation. Our QA program includes file checks, monthly checks (leaf position accuracy and interlock tests), and annual review. RESULTS: We found the MLC leaf position (light field) corresponds to decrement lines ranging from 50 to 59%. Transmission through MLC (1.5-2.5%) is less than alloy (3.5%). Multileaf penumbra is slightly wider than for alloy. Relative penumbra did not increase in the lung, and composite field dosimetry exhibited negligible differences compared with alloy. Verification simulations provide diagnostic image quality hard copies of the MLC fields. Monitor unit parameters used for alloy held for MLC. DISCUSSION: Clinical implementation for MLC as a block replacement was conducted on a site-by-site basis. Time studies indicate significant (25%) in-room time reductions. Through imaging and dosimetric analysis, the accuracy of field delivery has increased with MLC. The most significant impact of MLC is the ability to increase the number of daily treatment fields, thereby reducing normal tissue dosing, which is vital for dose escalation. PMID- 7493846 TI - Comparison of static conformal field with multiple noncoplanar arc techniques for stereotactic radiosurgery or stereotactic radiotherapy. AB - PURPOSE: Compare the use of static conformal fields with the use of multiple noncoplanar arcs for stereotactic radiosurgery or stereotactic radiotherapy treatment of intracranial lesions. Evaluate the efficacy of these treatment techniques to deliver dose distributions comparable to those considered acceptable in current radiotherapy practice. METHODS AND MATERIALS: A previously treated radiosurgery case of a patient presenting with an irregularly shaped intracranial lesion was selected. Using a three-dimensional (3D) treatment planning system, treatment plans using a single isocenter multiple noncoplanar arc technique and multiple noncoplanar conformal static fields were generated. Isodose distributions and dose volume histograms (DVHs) were computed for each treatment plan. We required that the 80% (of maximum dose) isodose surface enclose the target volume for all treatment plans. The prescription isodose was set equal to the minimum target isodose. The DVHs were analyzed to evaluate and compare the different treatment plans. RESULTS: The dose distribution in the target volume becomes more uniform as the number of conformal fields increases. The volume of normal tissue receiving low doses (> 10% of prescription isodose) increases as the number of static fields increases. The single isocenter multiple arc plan treats the greatest volume of normal tissue to low doses, approximately 1.6 times more volume than that treated by four static fields. The volume of normal tissue receiving high (> 90% of prescription isodose) and intermediate (> 50% of prescription isodose) doses decreases by 29 and 22%, respectively, as the number of static fields is increased from four to eight. Increasing the number of static fields to 12 only further reduces the high and intermediate dose volumes by 10 and 6%, respectively. The volume receiving the prescription dose is more than 3.5 times larger than the target volume for all treatment plans. CONCLUSIONS: Use of a multiple noncoplanar conformal static field treatment technique can significantly reduce the volume of normal tissue receiving high and intermediate doses compared with a single isocenter multiple arc treatment technique, while providing a more uniform dose in the target volume. Close conformation of the prescription isodose to the target volume is not possible using static uniform conformal fields for target shapes lacking an axis of rotational symmetry or plane of mirror symmetry. PMID- 7493845 TI - Conformal radiation therapy with fixed shaped coplanar or noncoplanar radiation beam bouquets: a possible alternative to radiosurgery. AB - PURPOSE: Three-dimensional (3D) geometric conformation of the therapeutic dose volume to the shape of a target tissue volume is the motivation for both conformal radiotherapy and radiosurgery. Although noncoplanar arcs have a clear physical and geometric advantage over fixed fields for small spherical targets, those advantages are reduced for large or irregularly shaped targets where static fields can be individually shaped. We have developed a system that allows efficient and flexible design and reliable delivery of customized "bouquets" of fixed nonopposed coplanar or noncoplanar shaped fields, resulting in highly uniform dose distributions. This report describes our initial experience using beam bouquets to treat intracranial lesions. METHODS AND MATERIALS: Patients with primary (11) or metastatic (4) intracranial lesions with a maximum diameter less than approximately 6 cm, most of whom candidates for single-fraction radiosurgery, were treated with beam bouquets of four to eight nonopposed coplanar or noncoplanar beams. Doses ranged from 16-20 Gy in four fractions for recurrent lesions (8) to 45 to 68 Gy in 25 to 34 fractions for primary lesions (7). The patients were immobilized with custom foam head supports and face masks attached to a fixed base plate. Planning computed tomography scans were acquired, from which the physician developed the custom beam bouquet using 3D treatment planning tools. The bouquet was designed based primarily on geometric concerns. The bouquet was subsequently modified to add wedge filters chosen by vector analysis of dose gradients to achieve uniform dose over the volume of beam crossfire. At the time of treatment, the isocenter was placed using the instructions provided by the treatment-planning system and pretreatment orthogonal port films were compared to digitally reconstructed radiographs (DRR) to assure proper isocenter placement. For several situations, the 3D dose distributions resulting from alternative coplanar and noncoplanar plans were compared. RESULTS: Each patient was treated without incident. Daily pretreatment port films showed excellent reproducibility of isocenter placement in 87% of setups. With short follow-up (0-12 months), two patients with recurrent glioblastoma experienced clinical deterioration 2 to 4 weeks following treatment. One had increased edema on scans and responded to steroids. Six patients clinically improved following radiation therapy. Review of alternative treatment plans reveals that the relative utility of coplanar vs. noncoplanar beams is likely dependent on the location of the lesion. Noncoplanar beam bouquets are likely preferable to coplanar beams when the target is located in the central regions of the head. Coplanar beams are likely adequate, and possibly preferable, for peripherally located targets. CONCLUSION: The biological advantages of fractionation and the physical advantages of radiosurgery are exploited with this approach. The use of multiple nonopposed coplanar or noncoplanar conformal wedged fields provides a uniform dose to the target and acceptable dose gradient at the target edge. This technique may prove to be an alternative to arc-based radiosurgery in some settings and has the potential advantages that fractionation should improve the therapeutic ratio, and each beam can be individually shaped to conform to irregularly shaped targets. Additional studies are underway to improve this system and better define its utility. PMID- 7493847 TI - Dosimetric evaluation of the conformation of the multileaf collimator to irregularly shaped fields. AB - PURPOSE: The goal of this study was to evaluate the dosimetric characteristics of geometric MLC prescription strategies and compare them to those of conventional shielding block. METHODS AND MATERIALS: Circular fields, square fields, and 12 irregular fields for patients with cancer of the head and neck, lung, and pelvis were included in this study. All fields were shaped using the MLC and conventional blocks. A geometric criterion was defined as the amount of area discrepancy between the MLC and the prescription outline. The "least area discrepancy" (LAD) of the MLC conformation was searched by selecting the collimator angle, meanwhile keeping a preselected position along the width of the leaf into the prescribed field. Five LAD conventions were studied. These included the LAD-0, LAD-1/3, LAD-1/2, and LAD-2/3 that inserted the leaves at the 0, 1/3, 1/2, and 2/3 of the leaf end into the prescription field, respectively. In addition, the LAD optimization was applied to the transecting (TRN) approach for leaf conformation that prescribed an equal area of overblocking and underblocking under each leaf. Film dosimetry was performed in a 20 cm polystyrene phantom at 10 cm depth 100 cm from source to axis distance (SAD) for both 6 and 18 MV photons with each of the above MLC conformations and conventional blocks. The field penumbra width, defined as the mean of the separation between the 20% and 80% isodose lines along the normal of the prescription field edge, was calculated using both the MLC and conventional block film dosimetry and compared. In a similar way, the d20 is defined as the mean separation between the 20% isodose line and the prescription field edge, and the d80 is defined as the mean separation between the 80% isodose line and the prescription field edge. RESULTS: The field penumbra width for all MLC conventions was approximately 2 mm larger than that of the conventional block. However, there was a larger variation of the separation distribution in the penumbra region of the irregular fields for the MLC, which had a standard deviation of 1 mm (a factor of 5 larger than the conventional block). The dosimetry for the circular fields showed that the LAD TRN, LAD-1/2, and LAD-1/3 approximated the conventional blocking well in terms of d20 and d80; however, no single convention produced the best conformation for both measures. The dosimetric result of the patient treatment fields was similar for all sites. The LAD-1/3, LAD-1/2, and LAD-TRN strategies conformed to within 1 to 1.5 mm of the d80 of the conventional block for both 6 MV and 18 MV, respectively. The LAD-1/2 and LAD-TRN conformations were virtually identical, although it is proven analytically that the LAD-1/2 convention has the least overall area discrepancy of all conventions. CONCLUSIONS: The five MLC conformation conventions resulted in similar dosimetric penumbrae for all field shapes studied. The LAD-1/3, LAD-TRN, and LAD-1/2 produced the more favorable approximation to conventional block. The field penumbra width, although useful for evaluating irregular field shapes, could not describe the large local variations in the penumbra along the field edge for the MLC. These local variations could be of clinical concern when they appear near vital organs. However, the variation in a local region can potentially be reduced by minimizing the jaggedness of the leaf steps in that local region. The dosimetric results were useful as guidelines for the clinicians in the evaluation and adjustment of MLC leaf positions. PMID- 7493848 TI - A precision cranial immobilization system for conformal stereotactic fractionated radiation therapy. AB - PURPOSE: Conformal radiotherapy has been shown to benefit from precision alignment of patient target to therapy beam (1, 6, 13). This work describes an optimized immobilization system for the fractionated treatment of intracranial targets. A study of patient motion demonstrates the high degree of immobilization which is available. METHODS AND MATERIALS: A system using dental fixation and a thermoplastic mask that relocates on a rigid frame is described. The design permits scanning studies using computed tomography (CT) and magnetic resonance imaging (MR), conventional photon radiotherapy, and high precision stereotactic proton radiotherapy to be performed with minimal repositioning variation. Studies of both intratreatment motion and daily setup reliability are performed on patients under treatment for paranasal sinus carcinoma. Multiple radiographs taken during single treatments provide the basis for a three-dimensional (3D) motion analysis. Additionally, studies of orthogonal radiographs used to setup for proton treatments and verification port films from photon treatments are used to establish day to day patient position variation in routine use. RESULTS: Net 3D patient motion during any treatment is measured to be 0.9 +/- 0.4 mm [mean +/- standard deviation (SD)] and rotation about any body axis is 0.14 +/- 0.67 degrees (mean +/- SD). Day-to-day setup accuracy to laser marks is limited to 2.3 mm (mean) systematic error and 1.6 mm (mean) random error. CONCLUSION: We conclude that the most stringent immobilization requirements of 3D conformal radiotherapy adjacent to critical normal structures can be met with a high precision system such as the one described here. Without the use of pretreatment verification, additional developments in machine and couch design are needed to assure that patient repositioning accuracy is comparable to the best level of patient immobility achievable. PMID- 7493849 TI - Effects of treatment setup variation on beam's eye view dosimetry for radiation therapy using the multileaf collimator vs. the cerrobend block. AB - PURPOSE: The purpose of this study is to quantify and compare retrospectively the effects of treatment setup variation on beam's eye view (BEV) dosimetry for radiation therapy using a multileaf collimator (MLC) vs. cerrobend block. METHODS AND MATERIALS: A study was performed on a group of 18 patients with cancer of the head and neck, lung, and pelvis who were treated with irregularly shaped fields. The BEV dosimetry of the fields shaped with cerrobend blocks and the MLC was measured with films at the depth of dose prescription in a solid water phantom. A "one-half-leaf" insertion convention was used to shape the MLC. In addition, an average of 15 sequential daily port films was taken per patient during the course of radiotherapy. The port films were aligned with the prescription film for each patient. Systematic error and random error of treatment setup for each patient were calculated. The effects of setup variation were incorporated by convolving the patient portal imaging data with the corresponding BEV film dosimetry. Two parameters were used to quantify the BEV dosimetry. First, the field penumbra width was calculated, which represented the average of the normal separations between 20 and 80% isodose lines along the prescription outline. Second, the ratio of areas covered by the 90 and 20% isodose lines, A90/20, was determined. The BEV dosimetry was then characterized with and without the effects of treatment setup variation. In addition, the difference in BEV dosimetry between the cerrobend block and the MLC was used to estimate the corresponding changes in tumor control probability (TCP). These changes were also compared to the changes in TCP for the treatment with or without the effects of random setup variation. RESULTS: With or without daily setup variation, the use of cerrobend block was more favorable than the MLC in terms of the field penumbra width and A90/20 for all treatment sites. In the absence of daily variation, the MLC field penumbra width was on average 1.3 mm larger than that of the cerrobend block, and 0.9 mm larger in the presence of daily setup variation. Similarly, the ratio A90/20 of the cerrobend block was on average 0.03 larger than that of MLC without daily setup variation, and 0.02 with daily setup variation. The difference in field penumbra width and A90/20 between the MLC and the cerrobend block was slightly reduced due to the effects of daily setup variation. For both the cerrobend block and the MLC, daily setup variation produced a significant increase in the field penumbra width, 2.3 mm for the cerrobend block and 1.9 mm for the MLC, and a decrease in the A90/20, 0.06 for the former and 0.05 for the latter. The change due to the daily setup variation was about a factor of 2 larger than the changes due to replacing the cerrobend block with the MLC. Using the TCP model, the change in TCP due to the daily setup variation was more than a factor of 3 larger than the change in TCP due to replacing the cerrobend block with the MLC. It was noted that the average changes in the penumbra, the A90/20 and the TCP calculated for the patient population did not adequately describe the changes for the individual patient. CONCLUSION: Our results do not show significant dosimetric differences between the MLC and the cerrobend block in conventional radiation treatment, whether or not daily setup variation was taken into consideration. The effects of daily setup variation alone produced a larger dosimetric change. The same results were obtained when the data were applied to calculate changes in TCP. For optimal radiation therapy, efforts should be concentrated on reducing daily setup variation. Our results also demonstrate the importance of frequent evaluation of MLC treatment using electronic portal imaging devices. PMID- 7493850 TI - Intraoperative optimization of needle placement and dwell times for conformal prostate brachytherapy. AB - PURPOSE: Traditionally, transperineal prostate brachytherapy has been heavily operator dependent. To overcome this limitation, a treatment planning method was developed for intraoperative planning, guidance, and evaluation. In this setting, reliability, speed, and ease of understanding are primary considerations. This planning method has been implemented for ultrasound guided implants of the prostate, but can be extended for use in other body sites. METHODS AND MATERIALS: The length and cross-section of the target (prostate) and location of urethra and rectum are determined intraoperatively from live ultrasound imaging. The planning program then automatically generates a "reference plan" containing needle locations, dwell times, and the resulting isodose distribution. As needles are placed, this information is corrected to account for any deviation of needle placement or movement of the prostate. Once all needles are in place, the normalization is adjusted to reconcile remaining hot-spots with coverage of the target volume. Optimization is performed in three separate stages. Each stage works to enhance only a subset of the implant parameters. (a) Pattern Optimization attempts to find the most appropriate placement for the needles or catheters. It is based on the transverse contour of the target volume. Needles are placed uniformly around the perimeter, and interior needle positions are determined from the cross-sectional area and shape. Critical structures such as the urethra are explicitly avoided. This step provides the overall framework for the implant, and is not generally repeated. (b) Relative Dwell Time Optimization selects relative dwell times that will give the best uniformity of dose. It works by setting the relative dwell time in each source position inversely proportional to the dose delivered to that point by the other source positions. It is used in the reference plan, and is repeated as each needle is inserted. This provides dosimetric feedback to the physician, who can judge the effect of deviations from the reference plan. (c) Relative Volume Optimization is an interactive method for fine tuning the normalization based upon volume analysis. The volume analysis is presented in tabular and graphical form, both being updated rapidly as the normalization is adjusted. The information is formatted to help the operator judge coverage and uniformity. Special functions are provided that allow the operator to "jump" to special normalization values based on several indices of uniformity or uniformity/coverage. RESULTS: This system overcomes some conventional brachytherapy limitations. Rather than depend on the operator's intuitive judgement of where the needles should be placed, a global plan is generated and validated with full dose calculations. Immediate feedback is provided concerning the adequacy of placement and avoidance of critical structures. This information is provided in terms of actual tissue doses to the target volume and critical structures using point doses, isodose distributions, and volume analysis. Since the new method was introduced in January 1994, 33 implants have been performed. The needle placement method has been reliable in the clinic, with different doctors producing similar results on subsequent fractions for the same patient. CONCLUSION: The method of decomposing the optimization problem into several simple steps is capable of rapidly, consistently, and reliably designing conformal treatment plans of high uniformity. Operator dependence has been significantly reduced. We are adapting the method for other anatomic sites. PMID- 7493851 TI - Dosimetric characteristics of a liquid-filled electronic portal imaging device. AB - PURPOSE: To determine the characteristics of a commercial electronic portal imaging device (EPID), based on a two-dimensional matrix of liquid-filled ionization chambers, for transmission dose measurements during patient treatment. METHODS AND MATERIALS: Electronic portal imaging device measurements were performed in a cobalt-60 beam and two accelerator x-ray beams, and compared with measurements performed with a Farmer-type ionization chamber in air in a miniphantom and in an extended water phantom. RESULTS: The warming up time of the EPID is about 1 h. The long-term stability of the detector is better than 1% under reference conditions for a period of about 3 months. The signal of the ionization chambers follows approximately the square root of the dose rate, although the relation becomes more linear for larger (> 1 Gy/min) dose rates. The signal can be transformed to dose rate with an accuracy of 0.6% (1 SD). The short term influence of integrated dose on the sensitivity of the ionization chambers is small. The sensitivity increases about 0.5% for all ionization chambers after an absorbed dose of 8 Gy and returns to its original value in less than 5 min after stopping the irradiation. This small increase in sensitivity can be ascribed to the electrode distance of the ionization chambers in commercial EPIDs, which is 0.8 +/- 0.1 mm. The sensitivity increase depends on the electrode distance and is 4% for a 1.4 mm electrode distance. The scattering properties of the EPID ionization chambers were between those of an ionization chamber in a miniphantom and in a water phantom. CONCLUSION: The matrix ionization chamber EPID has characteristics that make it very suitable for dose rate measurements. It is therefore a very promising device for in vivo dosimetry purposes. PMID- 7493852 TI - Dual-beam imaging for online verification of radiotherapy field placement. AB - PURPOSE: Due to the poor quality of megavoltage (MV) radiographs, detection and assessment of discrepancies in radiation field placement are difficult. Furthermore, the high imaging dose required to produce the megavoltage radiograph prohibits frequent image acquisition, particularly for those fields that require the use of an "open-field" exposure. For these small, or conformal, radiation fields, an alternate method of verifying field placement is required if the out of-field dose is to be minimized. An open-field image acquired with a kilovoltage (kV) source would (a) deliver a very low patient dose, (b) increase the visibility of bony landmarks, and (c) simplify intercomparison of portal and prescription images. This article describes the development of a dual-beam imaging system that produces diagnostic quality "double-exposure" portal images for verifying radiation field placement. METHODS AND MATERIALS: The dual-beam system consists of a kV x-ray tube mounted on the gantry of a medical linear accelerator. The kV beam shares the same isocenter (+/- 1 mm) as the treatment beam but is at 45 degrees to the central axis. Both the kilovoltage and megavoltage images are collected with a fluoroscopic imaging system that uses a low-noise CCD camera to accumulate the light emitted from a phosphor screen. Two 45 degrees mirrors are used to remove the CCD camera from the x-ray beam. The light integration on the CCD array is controlled by a mechanical shutter, allowing easy synchronization with the radiation exposures. The camera is shielded by a lead housing to reduce the number of x-rays reaching the CCD array. A conventional thickness phosphor screen is used for both the kV and MV exposures. In the dual-beam imaging procedure, an open-field kV radiograph is acquired with the patient in treatment position. Immediately following, a MV image is acquired with the beam-defining blocks in position. Summation of the two images produces an online double-exposure image. The anatomical information in either the kV or MV image can be emphasized by weighting the images appropriately. This system was used to acquire MV and kV images of both a contrast-detail phantom and a Rando head phantom. Dual-beam images were also acquired for a pituitary treatment, demonstrating the feasibility and usefulness of the dual-beam technique. RESULTS: Analysis of the contrast-detail images produced with the MV and kV beams shows the expected advantage of using the kV x ray beam. Images of a Rando head phantom confirm these results. A clinical demonstration of the dual-beam system for verifying the delivery of a pituitary field is shown. The quality of the dual-beam image is similar to the prescription (simulation) image, contains a larger anatomical region, and delivers a lower integral dose to the patient. In addition, the kV beam also enhances the visibility of small markers implanted in the prostate. CONCLUSIONS: A dual-beam imaging system has been developed for the radiographic verification of small, conformal fields. This development demonstrates the advantages and feasibility of using a kV x-ray beam in combination with the treatment beam to improve the accuracy of detecting patient setup errors. PMID- 7493853 TI - Automated localization of the prostate at the time of treatment using implanted radiopaque markers: technical feasibility. AB - PURPOSE: Prostate movement is a major consideration in the formation of target volumes for conformal radiation therapy of prostate cancer. The goal of this study was to determine the technical feasibility of using implanted radiopaque markers and digital imaging to localize the prostate at the time of treatment, thus allowing for reduction of the margin required for uncertainty in target position. METHODS AND MATERIALS: Radiopaque markers implanted around the prostate prior to treatment are visible on electronic radiographs generated with a portal imager or diagnostic imaging device. The locations of the images of these markers on the digital radiographs were automatically determined by a template-matching algorithm. The coordinates of the markers were found by projecting rays through the marker locations on orthogonal radiographs using a three-dimensional (3D) point-matching algorithm. Prostate and/or patient movement was inferred from the marker displacements. Images generated from known movements of a phantom with implanted markers were tested with this algorithm. Locations of markers from daily images of patients with implanted markers were determined by both manual and automatic techniques to determine the efficacy of automated localization on typical clinical images. RESULTS: Prostate movements can be automatically detected in a phantom using low-energy photons within 30 s after image acquisition and with a precision of better than 1 mm in translation and 1 degree in rotation (indistinguishable from the uncertainty in measuring precision). CONCLUSION: The studies show that on-line repositioning of the patient based on localization of the markers at the time of treatment is feasible, and may reduce the uncertainty in prostate location when combined with practical on-line repositioning techniques. PMID- 7493854 TI - Core-based portal image registration for automatic radiotherapy treatment verification. AB - PURPOSE: Portal imaging is the most important quality assurance procedure for monitoring the reproducibility of setup geometry in radiation therapy. The role of portal imaging has become even more critical in recent years due to the migration of three-dimensional (3D) treatment planning technology, including high precision conformal therapy, from the research setting to routine clinical practice. Unfortunately, traditional methods for acquiring and interpreting portal images suffer from a number of deficiencies that contribute to the well documented observation that many setup errors go undetected, and some persist for a clinically significant portion of the prescribed dose. Significant improvements in both accuracy and efficiency of detecting setup errors can, in principle, be achieved by using automatic image registration for on-line screening of images obtained from electronic portal imaging devices (EPIDs). METHODS AND MATERIALS: This article presents recent developments in a method called core-based image analysis that shows great promise for achieving the desired improvements in error detection. Core-based image analysis is a fundamental computer vision method that is capable of exploiting the full power of EPIDs by providing for on-line detection of setup errors via automatic registration of user-selected anatomical structures. We describe a robust method for automatic portal image registration based on core analysis and demonstrate an approach for assessing both accuracy and precision of registration methods using realistic, digitally reconstructed portal radiographs (DRPRs) where truth is known. RESULTS: Automatic core-based analysis of a set of 20 DRPRs containing known, random field positioning errors was performed for a patient undergoing treatment for prostate cancer. In all cases, the reported translation was within 1 mm of the actual translation with mean absolute errors of 0.3 mm and standard deviations of 0.3 mm. In all cases, the reported rotation was within 0.6 degree of the actual rotation with a mean absolute error of 0.18 degree and a standard deviation of 0.23 degree. CONCLUSION: Our results, using digitally reconstructed portal radiographs that closely resemble clinical portal images, suggest that automatic core-based registration is suitable as an on-line screening tool for detecting and quantifying patient setup errors. PMID- 7493855 TI - The cumulative verification image analysis tool for offline evaluation of portal images. AB - PURPOSE: Daily portal images acquired using electronic portal imaging devices contain important information about the setup variation of the individual patient. The data can be used to evaluate the treatment and to derive correction for the individual patient. The large volume of images also require software tools for efficient analysis. This article describes the approach of cumulative verification image analysis (CVIA) specifically designed as an offline tool to extract quantitative information from daily portal images. METHODS AND MATERIALS: The user interface, image and graphics display, and algorithms of the CVIA tool have been implemented in ANSCI C using the X Window graphics standards. The tool consists of three major components: (a) definition of treatment geometry and anatomical information; (b) registration of portal images with a reference image to determine setup variation; and (c) quantitative analysis of all setup variation measurements. The CVIA tool is not automated. User interaction is required and preferred. Successful alignment of anatomies on portal images at present remains mostly dependent on clinical judgment. Predefined templates of block shapes and anatomies are used for image registration to enhance efficiency, taking advantage of the fact that much of the tool's operation is repeated in the analysis of daily portal images. RESULTS: The CVIA tool is portable and has been implemented on workstations with different operating systems. Analysis of 20 sequential daily portal images can be completed in less than 1 h. The temporal information is used to characterize setup variation in terms of its systematic, random and time-dependent components. The cumulative information is used to derive block overlap isofrequency distributions (BOIDs), which quantify the effective coverage of the prescribed treatment area throughout the course of treatment. Finally, a set of software utilities is available to facilitate feedback of the information for treatment plan recalculation and to test various decision strategies for treatment adjustment. CONCLUSIONS: The CVIA tool provides comprehensive analysis of daily images acquired with electronic portal imaging devices. Its offline approach allows characterization of the nature of setup variation for the individual patient that would have been difficult to deduce using only a few daily or weekly portal images. Distribution of the tool will help establish an important database of setup variation from many clinics. The information derived from CVIA can also serve as the foundation to integrate treatment verification, treatment planning, and treatment delivery. PMID- 7493856 TI - Quantification of organ motion during conformal radiotherapy of the prostate by three dimensional image registration. AB - PURPOSE: Knowledge about the mobility of organs relative to the bony anatomy is of great importance when preparing and verifying conformal radiotherapy. The conventional technique for measuring the motion of an organ is to locate landmarks on the organ and the bony anatomy and to compare the distance between these landmarks on subsequent computerized tomography (CT) scans. The first purpose of this study is to investigate the use of a three dimensional (3D) image registration method based on chamfer matching for measurement of the location and orientation of the whole organ relative to the bony anatomy. The second purpose is to quantify organ motion during conformal therapy of the prostate. METHODS AND MATERIALS: Four CT scans were made during the course of conformal treatment of 11 patients with prostate cancer. With the use of a 3D treatment planning system, the prostate and seminal vesicles were contoured interactively. In addition, bladder and rectum were contoured and the volume computed. Next, the bony anatomy of subsequent scans was segmented and matched automatically on the first scan. The femora and the pelvic bone were matched separately to quantify motion of the legs. Prostate (and seminal vesicle) contours from the subsequent scans were matched on the corresponding contours of the first scan, resulting in the 3D rotations and translations that describe the motion of the prostate and seminal vesicles relative to the pelvic bone. RESULTS: Bone matching of two scans with about 50 slices of 256 x 256 pixels takes about 2 min on a workstation and achieves subpixel registration accuracy. Matching of the organ contours takes about 30 s. The accuracy in determining the relative movement of the prostate is 0.5 to 0.9 mm for translations (depending on the axis) and 1 degree for rotations (standard deviations). Because all organ contours are used for matching, small differences in delineation of the prostate, missing slices, or differences in slice distance have only a limited influence on the accuracy. Rotations of the femora and the pelvic bone are quantified with about 0.4 degree accuracy. A strong correlation was found between rectal volume and anterior-posterior translation and rotation around the left-right axis of the prostate. Consequently, these parameters had the largest standard deviations of 2.7 mm and 4.0 degrees. Bladder filling had much less influence. Less significant correlations were found between various leg rotations and pelvic and prostate motion. Standard deviations of the rotation angles of the pelvic bone were less than 1 degree in all directions. CONCLUSIONS: Using 3D image registration, the motion of organs relative to bony anatomy has been quantified accurately. Uncertainties in contouring and visual interpretation of the scans have a much smaller influence on the measurement of organ displacement with our new method than with conventional methods. We have quantified correlations between rectal filling, leg motions, and prostate motion. PMID- 7493857 TI - Evaluation of changes in the size and location of the prostate, seminal vesicles, bladder, and rectum during a course of external beam radiation therapy. AB - PURPOSE: To document the size and location of the prostate, seminal vesicles, bladder, and rectum throughout the course of external beam radiotherapy. The frequency and range of motion of these organs are quantified. METHODS AND MATERIALS: Ten patients with localized carcinoma of the prostate had conventional simulation followed immediately by a treatment planning computed tomography scan (TPCT0). Once treatment was initiated, each patient had a weekly CT (TPCT1-N) before or after his daily treatment. Anatomical structures from CT were delineated on a computer workstation for analysis. The serial CT sets were spatially registered to the initial scan using image correlation software that brings into congruence the bony pelvis of the different scans. The location of the prostate, seminal vesicles, bladder, and rectum on subsequent scans were compared to TPCT0, as well as to each other. RESULTS: Prostate volumes were observed to vary by an average of +/- 10% during the course of radiation therapy, while the seminal vesicle volumes varied by as much as 100%. Bladder and rectal volumes varied by +/- 30%. Compared to TPCT0, movement of the prostate was demonstrated in all patients. Quantitation of the center-of-mass (CM) showed motion of less than 1 mm in the left-right direction, while motion ranging from 0 to +/- 1 cm was observed in the anterior-posterior and superior-inferior directions. The individual standard deviations of these motions varied from approximately 1-5 mm. These variations were correlated to changes in the dimensions of the bladder and rectum. CONCLUSIONS: Changes in the location of the prostate, seminal vesicles, and normal tissue volumes during the course of radiation therapy occur and have dosimetric consequences that may impact tumor control and normal tissue complication probabilities. Conformal therapy for prostate cancer will require the incorporation of knowledge of the anatomic relationships of these structures as a function of time. Therefore, these uncertainties must be taken into account when designing treatment plans and in considering dose escalation trials. PMID- 7493858 TI - The effects of out-of-plane rotations on two dimensional portal image registration in conformal radiotherapy of the prostate. AB - PURPOSE: Rotations of the patient out of the image plane can significantly degrade the accuracy of two-dimensional (2D) image registration. This study determines the magnitude of the geometric errors introduced by 2D image registration as a result of out-of-plane rotations, and analyzes the dosimetric effects of these errors. METHODS AND MATERIALS: The magnitude of the errors introduced by 2D registration were determined by comparing orthogonal view portal images of a rotated phantom to simulator reference images of the same phantom without rotation. Dosimetric effects were calculated for three-dimensional (3D) conformal prostate treatments by applying the registration errors to patient treatment plans. The calculations were performed using a modified version of the dose calculation software used in our Cancer Center for 3D treatment planning based on computed tomography (CT). A method to detect out-of-plane rotations, specific to pelvic treatments, is introduced that uses the relative displacement of the centers of gravity of the acetabula in lateral images. RESULTS: The inherent uncertainty in the registration algorithm was 0.6 +/- 0.5 mm in translation and 0.7 +/- 0.8 degree in rotation within the image plane. For a 2 degrees out-of-plane rotation, the errors increase to 2.3 +/- 1.0 mm and 1.2 +/- 1.1 degrees. In some clinically realizable treatment scenarios it was observed that the errors introduced by the registration procedure could result in an overdosing of the rectal wall. The method to detect out-of-plane rotations was found to have an accuracy of better than 1 degree for rotations of less than 10 degrees. CONCLUSIONS: The errors introduced to the patient position by 2D image registration have dosimetrically significant consequences for out-of-plane rotations of 2 degrees or more. However, when used in conjunction with the method to detect out-of-plane rotations, 2D registration software was found to cause insignificant dose errors and, thus, become a more reliable and accurate clinical tool. PMID- 7493859 TI - Implementation of three dimensional conformal radiation therapy: prospects, opportunities, and challenges. AB - PURPOSE: To briefly review scientific rationale of 3D conformal radiation therapy (3DCRT) and discuss the prospects, opportunities, and challenges in the implementation of 3DCRT. Some of these ideas were discussed during a workshop on "Implementation of Three-Dimensional Conformal Radiation Therapy" in April 1994 at Bethesda, MD, and others have been discussed elsewhere in the literature. METHODS AND MATERIALS: Local-regional control of cancer is an important component in the overall treatment strategy in any patient with cancer. It has been shown that failure to achieve local-regional control can lead to (a) an increase in chances of distant metastases, and (b) a decrease in the survival. In many disease sites, the doses delivered currently are inadequate to achieve satisfactory local tumor control rates; this is because in many sites, only limited doses of radiotherapy can be delivered due to the proximity of cancer to radiosensitive normal tissues. By conforming the radiotherapy beams to the tumor, doses to the tumors can be enhanced and doses to the normal tissues can be reduced. With the advances in 3DCRT, such conformation is possible now and is the rationale for using 3DCRT. However, a number of questions do remain that are not limited to the following: (a) What are the implications in terms of target volume definitions when implementing 3DCRT? (b) Are there some sites where research efforts can be focused to document the efficacy and cost effectiveness of 3DCRT? (c) How do we implement day-to-day 3DCRT treatment efficiently? (d) How do we transfer the technology from the university centers to the community without compromising quality? (e) What are all the quality assurance/quality improvement questions that need to be addressed and how do we ascertain quality assurance of 3DCRT? (f) Have we looked at cost-benefit ratios and quality of life (QOL) issues closely? RESULTS: There is a need for defining multiple target volumes: gross tumor volume, clinical target volume(s), and planning target volume(s). Such definitions should make implementation of 3DCRT more complex, yet will make high dose delivery a possibility. There are many sites in which single and multiinstitutional studies are ongoing that include prostate, lung, head and neck, and brain. In other areas, cooperative group trials are required because of the inability of single institutions to accrue enough patients to answer clinically relevant questions with statistical validity. Although implementation of 3DCRT will require multiple steps, these multiple steps can be brought into clinical practice gradually and one does not have to wait until all steps required for implementation of 3DCRT are available. In this respect, "3DCRT" should be used in a very broad sense, from beam's eye view blocking, use of multibeam dose distribution, use of dose-volume histograms in choosing alternative plans, noncoplanar beam arrangements, intensity modulation, inverse planning, to totally automated implementation of 3DCRT. To transfer the 3DCRT capabilities to the community from the University Centers, there is a necessity to develop quality assurance programs. RTOG and the Three-Dimensional Oncology Group are spearheading these efforts. Three-dimensional conformal radiation therapy has potential not only to improve local control and decrease toxicity, but also to improve the cost benefit ratio in the use of radiotherapy as well as in improving quality of life in patients with cancer. CONCLUSIONS: Achieving many potential benefits of 3DCRT (improvement in local control, decreasing toxicity, organs-function preservation, improvement in cost effectiveness) will require further physics-related and clinical research in carefully conceived and successfully completed future clinical trials. PMID- 7493860 TI - Three dimensional conformal radiation therapy in pediatric parameningeal rhabdomyosarcomas. AB - PURPOSE: We evaluated the utility of three dimensional (3D) treatment planning in the management of children with parameningeal head and neck rhabdomyosarcomas. METHODS AND MATERIALS: Five children with parameningeal rhabdomyosarcoma were referred for treatment at our radiation oncology center from May 1990 through January 1993. Each patient was evaluated, staged, and treated according to the Intergroup Rhabdomyosarcoma Study. Patients were immobilized and underwent a computed tomography scan with contrast in the treatment position. Tumor and normal tissues were identified with assistance from a diagnostic radiologist and defined in each slice. The patients were then planned and treated with the assistance of a 3D treatment planning system. A second plan was then devised by another physician without the benefit of the 3D volumetric display. The target volumes designed with the 3D system and the two-dimensional (2D) method were then compared. The dosimetric coverage to tumor, tumor plus margin, and normal tissues was also compared with the two methods of treatment planning. RESULTS: The apparent size of the gross tumor volume was underestimated with the conventional 2D planning method relative to the 3D method. When margin was added around the gross tumor to account for microscopic extension of disease in the 2D method, the expected area of coverage improved relative to the 3D method. In each circumstance, the minimum dose that covered the gross tumor was substantially less with the 2D method than with the 3D method. The inadequate dosimetric coverage was especially pronounced when the necessary margin to account for subclinical disease was added. In each case, the 2D plans would have delivered substantial dose to adjacent normal tissues and organs, resulting in a higher incidence of significant complications. CONCLUSIONS: 3D conformal radiation therapy has a demonstrated advantage in the treatment of sarcomas of the head and neck. The improved dosimetric coverage of the tumor and its margin for subclinical extensions may result in improvement in local control of these tumors. In addition, lowering of radiation dose to adjacent critical structures may help lower the incidence of adverse late effects in children. PMID- 7493862 TI - Trust offers advice on emergency planning for your practice. PMID- 7493861 TI - Preliminary results of a prospective trial using three dimensional radiotherapy for lung cancer. AB - PURPOSE: To evaluate the preliminary results of a prospective trial using three dimensional (3D) treatment for lung cancer. METHODS AND MATERIALS: Seventy patients with inoperable Stage I through IIIB lung cancer were treated with three dimensional thoracic irradiation with or without chemotherapy (35% received chemotherapy). Total prescribed dose to the tumor ranged from 60-74 Gy (uncorrected for lung density). All patients were evaluated for local control, survival, and development of pneumonitis. These parameters were evaluated in respect to and compared with three-dimensional parameters used in their treatment planning. RESULTS: With a minimum follow-up of 6 to 30 months, the 2-year cause specific survival rate for Stages I and II was 90% and 53% for Stage III (no difference between Stages IIIA and IIIB). Patients with local tumor control had a better 2-year overall survival rate (47%) than those with local failure (31%). Volumetrically heterogeneously calculated doses were important to the accurate delineation of dose-volume coverage as there was a wide range of discrepancies between a homogeneously prescribed point dose calculation and the heterogeneously calculated volume coverage of that prescription. High-grade pneumonitis was correlated with the location of the tumor with lower lobe tumors having a much higher risk than those with upper lobe tumors. A critical volume effect and threshold dose were apparent in the development of high-grade pneumonitis. CONCLUSIONS: Three-dimensional therapy for lung cancer has been practically implemented at the Mallinckrodt Institute of Radiology and shows promising results in our preliminary analysis. The incidence of high-grade pneumonitis, however, warrants careful selection of patients for future dose escalation. Future dose escalation trials in lung cancer should be directed to volumes that limit the amount of elective nodal irradiation. However, the volume of necessary elective nodal irradiation remains unknown and should be studied prospectively. Dose escalation trials are indicated and may be facilitated by smaller target volumes. PMID- 7493863 TI - What's next after E coli O 157:H7? PMID- 7493864 TI - Equipping Pakistan for the future. PMID- 7493865 TI - Onion powder in baby food may induce anemia in cats. PMID- 7493866 TI - What is your diagnosis? Gastric foreign body and small intestinal ileus in a dog. PMID- 7493867 TI - The hyperfiltration theory: progression of chronic renal failure and the effects of diet in dogs. PMID- 7493868 TI - Racetracks and the veterinarian. PMID- 7493869 TI - Radioiodine treatment of 524 cats with hyperthyroidism. AB - OBJECTIVE: To evaluate a protocol for subcutaneous radioiodine treatment of cats with hyperthyroidism in which the dose was determined on the basis of severity of the cat's clinical signs, thyroid tumor size, and magnitude of the serum thyroxine (T4) concentration. DESIGN: Prospective case series. ANIMALS: 524 cats with hyperthyroidism. PROCEDURE: A scoring system based on 3 factors (severity of clinical signs, size of the thyroid gland, and magnitude of the serum T4 concentration) was used to select the dose of radioiodine to be administered subcutaneously. RESULTS: On the basis of the scoring system, 310 (59%) cats were treated with a low dose of radioiodine (< 3.5 mCi; median, 3.0 mCi), 158 (30%) were treated with a moderate dose (3.5 to 4.4 mCi; median, 4.0 mCi), and 56 (11%) were treated with a high dose (> or = 4.5 mCi; median, 5.0 mCi). At time of discharge from the hospital, serum T4 concentration was still high in 80 (15.3%) cats, but by 6 months after administration of radioiodine, the serum T4 concentration had decreased to within or below reference range in all but 8 (1.5%) cats with persistent hyperthyroidism. Many cats had low serum T4 concentrations at some time after radioiodine treatment, but only 11 (2.1%) cats developed clinical and clinicopathologic features of hypothyroidism and required supplementation with L-thyroxine. Thirteen (2.5%) cats had a relapse of hyperthyroidism 1.1 to 6.5 years after initial radioiodine treatment. Overall, the response to treatment was considered good in 94.2% of the cats. Median survival time in the cats was 2.0 years; the percentage of cats alive after 1, 2, and 3 years of treatment was 89, 72, and 52%, respectively. CLINICAL IMPLICATIONS: Results of the study suggest that this method of dose estimation works well and that subcutaneous administration of radioiodine provides a safe and effective means of treating hyperthyroidism in cats. PMID- 7493870 TI - Evaluation of factors associated with development of calcium oxalate urolithiasis in cats. AB - OBJECTIVE: To evaluate dietary and environmental factors as potential risk factors for calcium oxalate urolithiasis in cats. DESIGN: Case-control study. ANIMALS: 84 cats with uroliths composed of at least 90% calcium oxalate and 258 age- and gender-matched control cats. PROCEDURE: Owners of cats with calcium oxalate urolithiasis and control cats were surveyed between November 1990 and August 1992. Owners completed a standard questionnaire administered during a single telephone interview. Data collected included information regarding signalment, environment, urination and defecation, diet, and medical history. RESULTS: Calcium oxalate uroliths tended to develop in middle- to older-aged, domestic shorthair cats of typical weight. A gender predilection was not detected. Factors associated with an increase in the risk of calcium oxalate urolithiasis in cats were feeding urine-acidifying diets, feeding a single brand of cat food without providing additional foods or table scraps, maintaining cats in an indoor-only environment, and being of the Persian breed. CLINICAL IMPLICATIONS: Control of diet and environment may help prevent calcium oxalate urolithiasis. PMID- 7493871 TI - Serologic responses of dogs naturally exposed to or vaccinated against Borrelia burgdorferi infection. AB - OBJECTIVE: To characterize the serologic responses of dogs naturally exposed to or vaccinated against Borrelia burgdorferi and to assess responses at intervals after antibiotic treatment. DESIGN: Prospective, controlled clinical trial. ANIMALS: 19 dogs of various breeds and ages with narrowly defined clinical criteria of limb/joint borreliosis and 10 control dogs of equivalent age were used to determine serologic responses following natural exposure to the organism. Eight seronegative dogs were used to determine serologic responses following vaccination. PROCEDURE: Serologic responses to B burgdorferi and recombinant outer surface protein (Osp)A, flagellin, and P39 were assessed by means of ELISA and western immunoblot. Passive protective activity was assessed by use of a mouse protection assay. RESULTS: Naturally exposed dogs were seropositive, but had variable ELISA titers and immunoblot profiles. Immunoblot analysis did reveal consistent reactions to flagellin, P39, and a 22 kd protein, but not to OspA. Antibody responses did not change appreciably up to 13 weeks after antibiotic treatment. Vaccinated dogs had strong reactions to OspA and OspB, but not to P39. CLINICAL IMPLICATIONS: Dogs with clinical borreliosis are seropositive and remain seropositive after antibiotic treatment, emphasizing that serologic testing is not a useful means of measuring clinical response. Serologic responses of infected dogs can be discriminated from those of vaccinated dogs by means of immunoblot analysis, and recombinant P39 is a potentially useful antigen for that purpose. PMID- 7493872 TI - Intraocular silicone prosthesis implantation in eyes of dogs and a cat with intraocular neoplasia: nine cases (1983-1994). AB - OBJECTIVE: To determine the outcome of implantation of an intraocular silicone prosthesis (ISP) in the eyes of dogs and cats with intraocular neoplasia. DESIGN: Retrospective case series. ANIMALS: Eight dogs and 1 cat with histologically confirmed intraocular neoplasia. PROCEDURE: Signalment, clinical signs before and after surgery, results of microscopic examination of eviscerated intraocular tissues, follow-up information, and necropsy findings, if available, were recorded for each animal. RESULTS: Five dogs and 1 cat had primary intraocular neoplasms. Two of the dogs developed regrowth of the neoplasm around the ISP 6 to 24 months after implantation, and the eyes were enucleated. The cat developed signs compatible with systemic metastasis 4 years after surgery and was euthanatized. Three dogs had multicentric or metastatic neoplasia involving the eye. Two of these dogs were euthanatized because of their systemic disease 1.5 to 7 months after ISP implantation. The third dog is alive without evidence of regrowth 3 years after surgery. CLINICAL IMPLICATION: Intraocular neoplasia is not an absolute contraindication to ISP implantation. PMID- 7493873 TI - Accuracy of clinicians in predicting site and type of lesion as well as outcome in horses with colic. AB - OBJECTIVE: To assess the ability of clinicians to predict the site and type of lesion as well as outcome in horses with colic. DESIGN: Prospective case study. SAMPLE POPULATION: 139 horses admitted for evaluation of signs of colic. PROCEDURE: Six interns and residents examined horses with colic and predicted the segment of intestine that was affected, the type of lesion, and whether the horse would survive to discharge. Accuracy of prediction of site and type of lesion and survival prediction was compared between the first and second halves of the year, using chi 2 analysis and 95% confidence intervals on sensitivity and specificity. chi 2 Analysis was used to assess accuracy between predicted site and type of lesion and intraoperative or necropsy findings and to assess accuracy between predicted survival and actual outcome. RESULTS: Significant association existed between predicted segment of affected intestine or type of lesion and intraoperative findings (P < 0.05). There was a significant association between predicted survival and outcome (P < 0.001). Accuracy of survival prediction improved significantly (P = 0.002) during the year. CLINICAL IMPLICATIONS: Clinicians can accurately predict horses with colic that will survive surgery on the basis of clinical impressions. The ability to predict those horses that will survive improves with training. PMID- 7493874 TI - Evidence for a high rate of false-positive results with the indirect fluorescent antibody test for Ehrlichia risticii antibody in horses. AB - OBJECTIVE: The original objective was to determine seroprevalence of Ehrlichia risticii antibody among horses in California. On the basis of the unexpected results of the survey, an investigation into the accuracy and reproducibility of results of the indirect fluorescent antibody (IFA) test for E risticii was carried out. DESIGN: Prospective, seroprevalence study. ANIMALS: Healthy horses (n = 655) and horses with clinical signs of equine monocytic ehrlichiosis (EME; n = 514) from various regions of California. PROCEDURE: The IFA test was performed. Results were compared with results of an ELISA and with results of western immunoblot analysis. RESULTS: Overall, 104 of 655 (15.9%) healthy horses had evidence of an antibody response. However, 84 of 514 (16.3%) horses with clinical signs of EME also had positive test results, and of the 8 seropositive diseased horses for which paired (acute and convalescent) samples had been submitted, only 1 had a rise in antibody titers between the acute and convalescent samples. Comparison of results for the IFA test, ELISA, and western immunoblot analysis revealed a high rate of false-positive results for the IFA test. Subsequent studies suggested that routine vaccination of horses with non-E risticii vaccines may have contributed to the false-positive reactions. CLINICAL IMPLICATIONS: The data failed to provide conclusive evidence of E risticii infection among California horses. Owing to the high percentage of false-positive test results, caution is advised when using the IFA test to diagnose EME in horses or to determine the necessity for E risticii vaccination. PMID- 7493875 TI - Use of antibiotic-impregnated polymethyl methacrylate for treatment of an open radial fracture in a horse. AB - An open, contaminated, comminuted fracture of the radius of a 500-kg adult horse was diagnosed. Antibiotic-impregnated polymethyl methacrylate implants were placed into the fracture site to help eliminate infection from the limb. Later in the course of treatment, lag screws were placed across the fracture to generate compression and healing. The fracture healed, and the horse was released to its owners. The implants were not removed, and only a mild lameness existed 90 days later. PMID- 7493876 TI - Double-plate fixation of comminuted fractures of the second phalanx in horses: 10 cases (1985-1993). AB - OBJECTIVE: To determine the technique used, and the outcome for, double-plate fixation of comminuted fractures of the second phalanx of horses. DESIGN: Retrospective analysis of medical records. ANIMALS: 10 horses with comminuted fractures of the second phalanx that were treated by use of double-plate fixation. PROCEDURE: Two 4- to 6-hole dynamic compression plates were abaxially placed to achieve arthrodesis of the proximal interphalangeal joint and fixation of the fracture of the second phalanx. RESULTS: Arthrodesis and fracture healing were detected in all horses. CLINICAL IMPLICATIONS: Horses with comminuted fractures of the second phalanx that are treated with double-plate fixation have an excellent prognosis for survival and use as broodstock, and have an increased potential for return to useful function. PMID- 7493877 TI - Evaluation of the benefits of the timing of pregnancy testing by transrectal palpation in dairy cattle. AB - OBJECTIVE: To determine the benefits that were associated with pregnancy testing by use of transrectal palpation during the period 30 to 65 days after unsuccessful insemination of dairy cows. DESIGN: Nonconcurrent, cohort study. ANIMALS: Cows in 15 dairy herds in the United States and Canada. PROCEDURE: Reproductive records of cows (n = 713) that did not calve within 294 days of first-service insemination and that had been evaluated for pregnancy 30 to 65 days after first-service insemination were examined. Records were analyzed to determine the day of parturition or date of culling and to determine if the probability of a cow being culled or the interval to parturition was related to the number of days after insemination that pregnancy testing was performed. RESULTS: For cows that calved more than 294 days after first-service insemination, the interval from first-service insemination until parturition was associated significantly with herd, season, and treatment on the day of pregnancy testing with prostaglandin F2 alpha or one of its analogues. Cows treated with prostaglandin F2 alpha on the day of pregnancy testing were less likely to be culled than nontreated cows. For cows pregnancy tested 30 to 65 days after insemination, each additional day after day 30 before pregnancy testing was performed resulted in an increase of 1.09 days in the interval until parturition. CLINICAL IMPLICATIONS: Pregnancy testing by means of transrectal palpation as soon as possible after day 30 after insemination can result in shorter calving intervals. PMID- 7493879 TI - Survival of Serpulina hyodysenteriae in an effluent lagoon. AB - OBJECTIVE: To determine the survival of Serpulina hyodysenteriae in an infected lagoon that received effluent from a confinement building housing swine dysentery infected swine. DESIGN: Prospective controlled trial. ANIMALS: 48 shedder swine inoculated with S hyodysenteriae and housed in the building drained by the lagoon; 18 clinically normal detector swine confined in a separate building. PROCEDURE: Shedder swine were inoculated with S hyodysenteriae by oral administration of 20 g of diced colon from swine infected with swine dysentery. The lagoon that received effluent from the building housing the shedder swine was assayed for S hyodysenteriae by providing lagoon effluent twice daily for 2 or 4 days to detector swine as their sole source of drinking water and by subsequently examining these swine for signs of swine dysentery. Smears from rectal swab specimens and sometimes fecal specimens were stained for detection of large spirochetes. Fecal and rectal swab specimens and colonic scraping specimens were examined for S hyodysenteriae by anaerobic microbial culture on blood agar containing 400 micrograms of spectinomycin/ml. All shedder swine were necropsied after removal from the confinement building, as were detector swine after developing diarrhea or after 42 days of monitoring. RESULTS: For the first 5 to 6 days after removal of swine dysentery-infected shedder swine from the confinement building, lagoon effluent from the building remained infective. Detector swine, given lagoon effluent as their drinking water for a 2-day period, developed clinical swine dysentery, and S hyodysenteriae was cultured from specimens from these swine. Swine dysentery did not develop in each group of 2 detector pigs given lagoon effluent as their sole source of drinking water on days 7 and 8, 9 and 10, 11 through 14, or 15 through 18 after removal of the infected shedder swine. Large spirochetes were not observed on microscopy of stained colonic scraping specimens, and S hyodysenteriae and Salmonella spp were not cultured from specimens from these detector swine after being monitored for 42 days. Serpulina hyodysenteriae or Salmonella spp were not cultured from samples of the lagoon effluent. CLINICAL IMPLICATIONS: Although many factors could influence the survivability of S hyodysenteriae in a lagoon, results suggested that a facility with an open gutter-flush system that housed swine dysentery-infected swine should remain idle for more than 5 to 6 days before repopulating with unexposed swine. PMID- 7493881 TI - Acting against anthrax. US military veterinarian helps Haitians and their animals. PMID- 7493878 TI - Evaluation of liver function tests in neonatal calves. AB - OBJECTIVE: To evaluate the efficacy of tests for hepatic disease in young calves. DESIGN: Prospective case-control study. ANIMALS: 28 clinically normal calves and 47 calves with histologically proven hepatic disease. PROCEDURE: Liver function tests and serum activity of liver-derived enzymes were determined on 28 clinically normal calves at birth and at 2 weeks of age. These values were compared with the results from 47 calves with hepatic disease verified by histologic examination. Upper limit of confidence interval was determined for the results on the clinically normal calves, and Student's t-test was used to identify significant differences in the data from calves of various age-groups. RESULTS: None of the results of the common tests for liver damage or function (measurement of bilirubin, gamma-glutamyltransferase, glutamate dehydrogenase [GMD], alkaline phosphatase, L-lactate dehydrogenase, aspartate transaminase, or alanine transaminase) were clinically useful when used alone for detection of hepatic disease in calves less than 6 weeks old. Sensitivity of gamma glutamyltransferase, GMD, aspartate transaminase, and alkaline phosphatase as indicators of hepatic disease in this population of calves was 0, 59, 80, and 9%, respectively. Direct bilirubin (sensitivity, 87%) was more sensitive than total bilirubin (sensitivity, 66%). Serum enzyme activity of most enzymes (P < 0.01), total bilirubin concentration (P < 0.001), and sulfobromophthalein sodium clearance half-life were significantly higher (P < 0.001) in newborn calves than in 2-week-old calves. CLINICAL IMPLICATIONS: Clinical findings that indicate hepatic disease in calves that are less than 6 weeks old could be confirmed by measurement of serum activity of GMD or concentrations of total serum bile acids or direct bilirubin. Percutaneous liver biopsy may still be needed and may provide the most information. PMID- 7493880 TI - Parasitisms of catfish and other farm-raised food fish. PMID- 7493882 TI - The battle against E coli; veterinarians play critical role. PMID- 7493883 TI - Legislative and regulatory front. A question of acclimation. PMID- 7493884 TI - Vietnamese potbellied pigs: myths and misconceptions. PMID- 7493885 TI - Proponent of training in aquatic animal health. PMID- 7493886 TI - Opposed to fee splitting. PMID- 7493887 TI - Thoughts on animal rights and animal liberation. PMID- 7493888 TI - Thoughts on animal rights and animal liberation. PMID- 7493889 TI - Clarification on firearms and ammunition. PMID- 7493891 TI - Opportunities for food animal practitioners to promote human and animal health. PMID- 7493892 TI - What is your diagnosis? Small intestinal obstruction in a cat. PMID- 7493890 TI - Description and evaluation of a course in veterinary ethics. PMID- 7493894 TI - Rabies surveillance in the United States during 1994. AB - In 1994, 48 states, the District of Columbia, and Puerto Rico reported 8,224 cases of rabies in nonhuman animals and 6 cases in human beings to the Centers for Disease Control and Prevention. Nearly 93% (7,632 cases) were wild animals, whereas 7% (592 cases) were domestic species. The total number of reported cases decreased 13.4% from that of 1993 (9,498 cases), with most of the decline resulting from 19.2% fewer cases of rabies in raccoons. Two previously described epizootics of rabies involving the raccoon variant of the rabies virus have converged in North Carolina, and the resulting region is now continuous from Alabama and Florida in the South to Maine in the North. Epizootics of rabies in foxes in west central Texas and in dogs and coyotes in southern Texas continue to expand, with this state reporting 144 rabid foxes, 53 rabid dogs, and 77 of the 85 cases in coyotes during 1994. Maine and New Hampshire reported cases of rabies in foxes (6 and 9, respectively) for the first time in 10 years. Nationally, reported cases of rabies in dogs (153) increased by 17.7%, whereas cases in cattle (111) and cats (267) decreased by 14.6 and 8.3%, respectively. Cats continued to be the domestic animal most frequently reported rabid. Twenty-eight states and the District of Columbia reported decreases in rabies in animals in 1994, compared with 22 states, the District of Columbia, and Puerto Rico in 1993. Hawaii and Nebraska were the only states that did not report cases of rabies in 1994. PMID- 7493893 TI - Employment of 1995 graduates of US veterinary medical colleges, analysis by gender. PMID- 7493895 TI - Clinical efficacy of increased dosages of milbemycin oxime for treatment of generalized demodicosis in adult dogs. AB - OBJECTIVE: To determine the efficacy of increased dosages of milbemycin oxime in the treatment of generalized demodicosis. DESIGN: Prospective clinical trial. ANIMALS: 26 adult dogs with chronic generalized demodicosis. PROCEDURE: In phase 1, milbemycin was administered daily to 13 dogs at an approximate mean dosage of 1 mg/kg of body weight (PO) until 30 days after skin scrapings failed to detect mites. If the mite count had not decreased by 25% from the prior month's examination, the drug dosage was increased to approximately 2 mg/kg. Treatment was considered a failure if the mite count had not changed on 2 successive examinations. In phase 2 involving 13 other dogs, an approximate mean dosage of 2 mg/kg was used. If mite counts had not been reduced to 0 by 180 days, treatment was considered to have failed. RESULTS: In phase 1, when milbemycin was administered at the initial low dosage, 6 dogs were considered to have been cleared of mites. One of these relapsed 2 months after discontinuation of treatment. For the 7 dogs not cleared of mites, the dosage was doubled. Two of these were never cleared of mites. In phase 2, 12 of 13 dogs were cleared of mites after 60 to 180 days of treatment. CLINICAL IMPLICATIONS: High-dose milbemycin was effective in the treatment of generalized demodicosis. PMID- 7493897 TI - Nasopharyngeal epidermal cyst in a dog. AB - Plain film radiography, magnetic resonance imaging, and histologic evaluation were used to diagnose nasopharyngeal epidermal cyst in a 12-year-old male Miniature Poodle with inspiratory stridor. The cyst was lined with stratified squamous epithelium, with variable keratinization. Most of the epithelium was well-differentiated and supported by a fibrovascular tissue separating it from underlying woven bone. One margin of the mass had a transition from stratified squamous to columnar respiratory epithelium. Other reported epidermal cysts in dogs have been confined to the middle ear or intracranial area. This case was unique, because the mass was located in the nasopharyngeal area, causing airway obstruction. This lesion was most likely acquired rather than congenital, because the dog had a history of middle ear inflammation, as well as bite wound trauma to the throat region. The mass was successfully removed by use of a transpalatal approach, and signs attributable to respiratory obstruction were alleviated. The dog had no abnormal clinical signs several months after the procedure. PMID- 7493896 TI - Inheritance of cystinuria and renal defect in Newfoundlands. AB - OBJECTIVE: To describe clinical features, characterize metabolic renal abnormalities, and evaluate mode of inheritance of cystinuria in Newfoundlands. DESIGN: Prospective study. ANIMALS: Two families of Newfoundlands including 11 dogs with dysuria, stranguria, or obstruction attributable to cystine calculi. PROCEDURE: Urinalysis and nitroprusside spot tests were performed to evaluate cystinuria in the affected dogs. All calculi were analyzed by crystallography. Amino acid concentrations in urine and plasma of affected dogs were compared with those in clinically normal related dogs. Renal fractional excretion and reabsorption of amino acids were determined in 5 affected Newfoundlands. RESULTS: Nine dogs had pure cystine calculi in the bladder, and 4 of these had renal cystine calculi. Affected dogs persistently excreted excessive amounts of cystine (> 500 nmol/mg of creatinine; reference = 54 +/- 38 nmol/mg of creatinine) and had typical cystine crystals in acidic urine. Urinary excretion of ornithine, lysine, and arginine was also high. Dogs with cystinuria had complete lack of reabsorption and active secretion of cystine, and reabsorption of lysine, ornithine, and arginine was moderately impaired. Although clinical signs of urinary obstruction were observed only in males, cystinuric male and female offspring were produced from noncystinuric parents, consistent with an autosomal recessive mode of inheritance. Obligate heterozygotes did not have clinical signs, and had normal urinary cystine content and renal amino acid reabsorption. CLINICAL IMPLICATIONS: Because detection of carriers by routine urinalysis is currently not possible, Newfoundlands with cystinuria and their parents and offspring should be excluded from breeding. PMID- 7493898 TI - Alimentary lymphoma in cats: 28 cases (1988-1993). AB - OBJECTIVE: To evaluate response to chemotherapy in cats with alimentary lymphoma and to determine factors associated with survival time. DESIGN: Retrospective case series. ANIMALS: 28 cats with alimentary lymphoma that underwent chemotherapy. RESULTS: In all cats, the diagnosis had been established by means of cytologic or histologic examination of ultrasound-guided aspirates and biopsy specimens (18 cats), histologic examination of surgically obtained biopsy specimens (7 cats), or examination of specimens obtained endoscopically (3 cats). Clinical signs included anorexia, weight loss, vomiting, and diarrhea. Twenty seven cats were treated with vincristine sulfate, cyclophosphamide, and prednisone; 1 was treated with chlorambucil and prednisone. Survival time ranged from 2 to 2,120 days (median, 50 days). Nine cats achieved complete remission (remission time ranged from 30 to 1,700 days; median, 213 days), 2 achieved partial remission, and 17 failed to respond to chemotherapy. Sex, FeLV status, hematocrit, serum total protein concentration, site and extent of gastrointestinal involvement, and clinical stage were not found to be associated with survival time. CLINICAL IMPLICATIONS: Cats with alimentary lymphoma are poorly responsive to treatment with vincristine, cyclophosphamide, and prednisone; however, a small subset of cats may have long survival times. PMID- 7493899 TI - Laparoscopic inguinal herniorrhaphy in two stallions. AB - Two mature stallions that were used for breeding purposes were admitted for evaluation of inguinal hernias. In 1 horse, the hernia was reduced per rectum by gentle traction applied to the intestine. In the other horse, the hernia was reduced by placing the horse in dorsal recumbency and applying external pressure over the scrotum. Both horses were observed for recurrence of the hernia. Seven days later, an elective laparoscopic technique for inguinal herniorrhaphy was performed on each horse, using polypropylene mesh. Complications did not develop, and both horses have successfully completed 2 full breeding seasons. Laparoscopic inguinal herniorrhaphy allows preservation of the testis on the affected side and precludes many complications associated with open surgical techniques used on inguinal rings. PMID- 7493901 TI - Evaluation of the pathogenesis of vagus indigestion in cows with traumatic reticuloperitonitis. AB - OBJECTIVE--To evaluate the functional importance of impaired reticular contractions attributable to inflammatory adhesions in the pathogenesis of vagus indigestion in cows with traumatic reticuloperitonitis (TRP). DESIGN--Case control study. ANIMALS--42 dairy cows with TRP were compared with 10 healthy control cows. Cows with TRP were assigned to 3 groups: cows with uncomplicated TRP (n = 15), cows with TRP and a disturbance of digesta passage through the reticular-omasal orifice (TRP-AD; n = 17), and cows with TRP and a disturbance of digesta passage through the pylorus (TRP-PD; n = 10). PROCEDURE--Cows were evaluated, using routine clinical methods, biochemical analysis of blood and ruminal fluid samples, exploratory rumenotomy, and postmortem examination. RESULTS--Damage of thoracic and abdominal parts of the main vagal branches were not evident in cows with TRP. Parareticular inflammatory adhesions were more extensive in cows with TRP-AD and in cows with TRP-PD than in cows with uncomplicated TRP. Reticular motility was decreased in cows with TRP-AD and TRP PD, compared with cows with uncomplicated TRP and control cows. In contrast to cows with uncomplicated TRP and cows with TRP-AD, the abomasum was impacted with large amounts of long fibrous material in most of the cows with TRP-PD. CLINICAL IMPLICATIONS--We believe that disturbances of digesta passage in cows with TRP represent distinct stages of a common pathogenetic course. The primary disease mechanism was a disturbance in particle-separating processes in the reticulum/rumen attributable to mechanical inhibition of reticular motility that was the result of extensive inflammatory parareticular adhesions. PMID- 7493902 TI - Evaluation of the use of intrauterine infusions of oxytetracycline, subcutaneous injections of fenprostalene, or a combination of both, for the treatment of retained fetal membranes in dairy cows. AB - Dairy cows (n = 207) that had retained fetal membranes for more than 8 hours after parturition were randomly assigned to 1 of 4 treatment groups (group 1, daily intrauterine infusion of oxytetracycline; group 2, daily intrauterine infusion of oxytetracycline and a single SC injection of fenprostalene; group 3, a single SC injection of fenprostalene; and group 4, untreated controls). Cows were monitored daily. Subsequent disease conditions were recorded, and appropriate treatments were initiated. Interval from parturition to expulsion of fetal membranes was unaffected by treatment regimen. The frequency of cows with high rectal temperatures (> or = 39.5 C) was significantly (P < 0.05) less in cows treated with oxytetracycline infusions; however, the frequency of displaced abomasum, ketosis, and mastitis was unaffected by treatment method. Treatment method appeared to influence the development of postpartum uterine pathologic conditions. Treatment with oxytetracycline and fenprostalene (group 2) increased (P < 0.05) the frequency of pyometra. Treatment with fenprostalene (groups 2 and 3) decreased the number of cows with a palpable uterine lumen detectable during per rectal palpation at 28 to 42 days after parturition. Reproductive efficiency, as determined on the basis of the interval from parturition to conception, was unaffected by treatment protocol. PMID- 7493903 TI - Rostral mandibulectomy for treatment of an ameloblastic fibro-odontoma in a cow. AB - A 1-year-old polled Hereford heifer was admitted with a mass of the rostral mandibular area that had been observed 17 days earlier. Cytologic examination of fluid aspirated from the mass revealed normal blood. Radiography revealed an expansile, osteolytic mass involving the rostral portion of the mandible. Mineralization of the soft-tissue component was evident. Rostral mandibulectomy was performed to remove the mass in its entirety. The histologic diagnosis was ameloblastic fibro-odontoma. One year after surgery, cosmetic appearance of the mandible was acceptable, and there was no evidence of tumor regrowth. Radiation therapy, cryotherapy, and thermocautery have been recommended in conjunction with surgical excision to prevent tumor regrowth in the oral cavity. Tumor regrowth was not a factor in long-term prognosis in this cow. PMID- 7493900 TI - Use of imipramine hydrochloride for treatment of urospermia in a stallion with a dysfunctional bladder. AB - An 8-year-old stallion was evaluated because of recurrent urinary tract infections and chronic intermittent urospermia. After extensive diagnostic testing, it was hypothesized that the stallion had a reflex dyssynergia of the bladder and urethral sphincter. Initial attempts to manage the urospermia included semen fractionation, semen collection after voluntary urination, and use of semen extenders. None of these efforts reliably yielded a quality ejaculate. Administration of imipramine hydrochloride (1.2 mg/kg of body weight, PO, 4 hours prior to semen collection) was initiated in an attempt to enhance bladder neck closure during ejaculation. This treatment, combined with voluntary urination prior to ejaculation, resulted in ejaculates containing little or no urine. Using this protocol, 19 of 20 mares bred during the subsequent 2 years became pregnant. By the third year, the bladder dysfunction had progressed, and the urospermia was no longer manageable. Bladder catheterization, followed by manual expression of the bladder per rectum, were necessary prior to each semen collection to obtain a urine-free ejaculate. Three-and-a-half years after initial examination, transitional cell carcinoma of the bladder with metastasis was identified, and the stallion was euthanatized. It is not known whether the transitional cell carcinoma was related to the dysfunctional bladder. Imipramine hydrochloride did not eliminate, but did reduce, the frequency and degree of urospermia in the affected stallion for approximately 2 years. PMID- 7493905 TI - Vision in dogs. AB - Compared with the visual system in human beings, the canine visual system could be considered inferior in such aspects as degree of binocular overlap, color perception, accommodative range, and visual acuity. However, in other aspects of vision, such as ability to function in dim light, rapidity with which the retina can respond to another image (flicker fusion), field of view, ability to differentiate shades of gray, and perhaps, ability to detect motion, the canine visual system probably surpasses the human visual system. This has made the dog a more efficient predator in certain environmental situations and permits it to exploit an ecological niche inaccessible to humans. PMID- 7493904 TI - Epizootic of Mycobacterium bovis in a zoologic park. AB - An epizootic of Mycobacterium bovis in a zoologic park resulted in the death of 4 southern white rhinoceroses and 2 colobus monkeys. Zoo personnel were detected that had positive intradermal tuberculin skin test results after exposure to mycobacterial-infected animals. On the basis of DNA fingerprinting, all 3 mycobacterial isolates (from 1 rhinoceros and 2 monkeys) were determined to be genetically similar and probably originated from the same source. The 3 animals (1 rhinoceros and 2 colobus monkeys) that had confirmed infections lived in separate, but adjacent, areas. Aerosolization of bacteria during routine cleaning was believed to have contributed to the unusual distance between infected animals. Tuberculosis has reemerged as a major disease problem in human and veterinary medicine. PMID- 7493906 TI - Restoration of the cellular senescence program and repression of telomerase by human chromosome 3. AB - Telomeres, at the end of chromosomes, shorten with each cell division, resulting in cellular senescence. Tumor cells, unlike normal somatic cells, express a telomerase that maintains the telomere length. Deletion of a gene(s) on chromosome 3 is common in human renal cell carcinoma (RCC) and reintroduction of a normal chromosome 3 into an RCC immortal cell line restored the program of cellular senescence. The loss of indefinite growth potential was associated with the loss of telomerase activity and shortening of telomeres in the RCC cells with a normal chromosome 3. However, microcell hybrids that escaped from senescence and microcell hybrids with an introduced chromosome 7 or 11 maintained telomere lengths and telomerase activity similar to those of the parental RCC23. Thus, restoration of the cellular senescence program by chromosome 3 is associated with repression of telomerase function in RCC cells. PMID- 7493908 TI - Effects of menstrual and reproductive factors on the risk of breast cancer: meta analysis of the case-control studies in Japan. AB - To elucidate the magnitude of the effect of menstrual and reproductive factors on breast cancer occurrence among Japanese women, we reviewed eight case-control studies previously conducted in Japan and used a quantitative method (meta analysis) to summarize the data. While individual studies have different methods and populations, the estimated odds ratios (ORs) in the studies were statistically homogeneous for all study variables. It was confirmed that early age at menarche, late age at first birth, and premenopausal status are significantly associated with risk of breast cancer; an estimated combined OR of 0.68 (95% confidence interval (CI): 0.59-0.77) was obtained for women with onset of menstruation after age 16 compared to those before age 14. Nulliparous women had higher risk than women with first birth before age 25 (OR = 1.56 95%, CI: 1.27-1.91). The OR for women with first birth after age 35 was 2.26 (95% CI: 1.85 2.77) compared to women at first birth before age 25. Premenopausal women had a higher risk than women with menopause before age 50 (OR = 2.21, 95% CI: 1.53 3.20). We also found a significant protective effect of high parity after controlling for age at first birth and the other menstrual factors. The OR estimate for 3 or more births compared to nulliparity was 0.68 (95% CI: 0.54 0.86). The meta-analysis provided quantitative estimates of breast cancer risk among Japanese women with improved precision. PMID- 7493907 TI - Cloning of the rat homologue of the von Hippel-Lindau tumor suppressor gene and its non-somatic mutation in rat renal cell carcinomas. AB - Recently, von Hippel-Lindau (VHL) gene mutations were detected in non-inherited, sporadic human renal cell carcinomas (RCs) at a high frequency. In order to determine whether or not the VHL gene is also a critical gene in rat RCs, we cloned and sequenced the rat homologue of human VHL gene and searched for mutations of the VHL gene in rat RCs. Mutations in the VHL gene were not detected in spontaneous RCs of the Eker rat model or in ferric nitrilotriacetate-induced rat RCs using the polymerase chain reaction-single strand conformation polymorphism (PCR-SSCP) method. These data indicate that mutation of the VHL tumor suppressor gene is not an event in rat renal carcinogenesis, at least in our present systems. PMID- 7493909 TI - Gastric cancer among the Japanese in Hawaii. AB - The incidence rate of gastric cancer among men of Japanese ancestry living in Hawaii is about one-third as high as that of their counterparts living in Japan. Because of this difference, a prospective study was conducted to identify factors related to the development of gastric cancer in Hawaii. Eight thousand and six (8,006) men born from 1900-1919 were examined from 1965 to 1968 and followed for over 25 years. During this time, 250 incident cases of gastric cancer were identified. The study has found the following: 1) prior infection with Helicobacter pylori bacteria increased the risk for stomach cancer; 2) cigarette smoking was positively associated with gastric cancer with age at which smoking started being an important risk factor; 3) after taking cigarette smoking into account, alcohol intake was not related to stomach cancer risk; 4) a low pepsinogen I level identified subjects at increased risk for the intestinal histologic type of gastric cancer; 5) a low serum ferritin level was a marker for increased risk of stomach cancer; 6) there was a weak indication that the intake of vegetables and fruits was inversely related to gastric cancer; 7) there was no association of stomach cancer with levels of serum cholesterol, serum uric acid, serum micronutrients (retinol, beta-carotene or alpha-tocopherol) or blood hematocrit; 8) there was also no association of gastric cancer with body mass index or physical activity. PMID- 7493910 TI - Progesterone enhancement of stomach tumor development in SD rats treated with N methyl-N'-nitro-N-nitrosoguanidine. AB - The effects of chronic progesterone treatment on gastric tumorigenesis were examined in 6-week-old male SD rats. The rats were castrated, progesterone or testosterone pellets were implanted, and, starting one week after the operation, 100 mg/liter of N-methyl-N'-nitro- N-nitrosoguanidine (MNNG) was administered in the drinking water for 16 weeks. Every 2 months the pellets were changed. Group 1 animals received castration plus MNNG while Groups 2 and 3 also received progesterone and testosterone, respectively. In the Group 4 case, progesterone and testosterone were administered alternately for 2-month periods and in Group 5 MNNG was given to intact animals. All survivors were killed one year after the start of MNNG treatment. In Group 1 the incidence of gastric tumors was significantly decreased as compared with the Group 5 value. The Group 2 incidence, in contrast, was similar to that in Group 5, and the size of the observed gastric tumors was massively increased. The area of the pyloric gland mucosa was also greater than in other groups. Testosterone treatment was associated with a less pronounced increase in tumor size and a recovery in incidence. The results indicate that progesterone may exert a promoting influence on gastric tumor development. PMID- 7493911 TI - Low incidence of a nucleotide sequence alteration of the neurofibromatosis 2 gene in human breast cancers. AB - We investigated aberrations of the neurofibromatosis type 2 (NF2) gene in breast tumors of 60 patients by single-strand conformation polymorphism analysis of polymerase chain reaction products followed by nucleotide sequencing. We detected a tumor-specific single-base substitution in codon 398 in exon 12 of the gene, resulting in an alteration of a single amino acid, in DNA from a breast cancer sample. The results indicated possible but infrequent involvement of mutations of the tumor suppressor NF2 gene in human breast cancers. PMID- 7493913 TI - Transformation of BALB/c 3T3 cells in vitro by the fungicides captan, captafol and folpet. AB - Cytotoxic and cell-transforming activities of the three fungicides, captan, captafol and folpet, have been studied in an experimental in vitro model by exposing BALB/c 3T3 cells to the chemicals with or without S-9 mix-induced bioactivation. Cytotoxicity of the three compounds was reduced in the presence of the metabolizing system. Each assayed pesticide displayed cell-transforming ability in the presence of the metabolizing system. The relative efficiency was: captafol > captan > folpet. Cell transformation was considered to be due to carcinogenesis-promoting activity. These data, obtained in a medium-term (6-8 weeks) experimental model, contribute to a better understanding of the action of the three pesticides in the multistep carcinogenesis process and provide more information concerning the oncogenic risk of these xenobiotic compounds for humans. PMID- 7493912 TI - Expression of integrin alpha 3 in gastric and colorectal cancers: its relation to wall contraction and mode of invasion. AB - We macroscopically classified 25 gastric and 23 colorectal advanced cancers into "contracted" and "uncontracted" types, and found immunohistochemically that integrin subunit alpha 3 was more frequently expressed in the extracellular matrix (ECM) in the former than in the latter (75%:9/12 vs. 38%: 5/13 in gastric and 86%:6/7 vs. 25%:4/16 in colorectal cancers, respectively). Integrin subunit alpha 3 was also expressed more frequently in cancers producing transforming growth factor-beta (TGF-beta), which is related to ECM deposition, integrin expression and cell mobility, than in those which did not produce TGF-beta (67%:10/15 vs. 40%:4/10 in gastric and 57%:4/7 vs. 38%:6/16 in colorectal cancers, respectively). In addition, integrin subunit alpha 3 was not expressed in 2 benign gastric ulcers combined with gastric cancer elsewhere in the stomach. On the other hand, a retrospective analysis of 107 cases of rectal cancer which recurred after a curative operation revealed that local recurrence was more frequent in "contracted" than "uncontracted" types (44%:11/25 vs. 26%:21/82). These results may suggest that the abundant interstitial fibrosis which leads to remarkable gastric or colorectal wall contraction is a result of the interaction between cancer cells and ECM, along with the expression of integrin and/or the production of TGF-beta. This fibrosis may also be closely related to the mode of gastric and colorectal cancer invasion. PMID- 7493914 TI - Induction of erythroid differentiation by 5-fluorouracil in K562 leukemia cells. AB - K562 cell line, established from a patient in the blast crisis of chronic myeloid leukemia, expresses high levels of c-myc and bcr/abl gene products. Exposure of K562 cells to 5-fluorouracil (5-FU) resulted in a marked benzidine-positive erythroid differentiation with concomitant reduction in cell proliferation. No change in c-myc mRNA or protein levels occurred during 96 h of drug treatment. In contrast, a biphasic change of p210bcr/abl and the abl-associated kinase activities was observed upon treatment with 5-FU. The change in p210bcr/abl expression may be mediated at the translational level, since the steady-state level and the enzymatic activity of p210bcr/abl are reduced, whereas bcr/abl mRNA levels are unaltered. The results are consistent with the existence of pleiotropic differentiation pathways in K562 cells. PMID- 7493916 TI - Analysis of naturally processed human histocompatibility leukocyte antigen class I-bound peptides from hepatocellular carcinoma tissues in vivo. AB - Naturally processed self-peptides bound to human histocompatibility leukocyte antigens (HLA) class I molecules of human hepatocellular carcinoma tissues (HLA A2.1, -B44, -B13) in vivo were isolated for sequence analysis. Acid-eluted peptides were subjected to reversed-phase high-performance liquid chromatographic separation and single-fraction sequencing was performed by Edman degradation. The peptides were found to be octamers or nonamers and they were derived from the processing of intracellular proteins. Three independent sequences were obtained from HLA-A2.1 molecules. One of the peptides showed sequence homology to the hepatitis B virus (HBV) pre-S protein, one to aldehyde dehydrogenase, and the other to no known protein. Two independent sequences were obtained from HLA-B44, B13 molecules: one showed sequence homology to the human c-abl protein, the other showed no homology to any known protein. A synthetic biotinylated peptide based on the HBV pre-S peptide sequence was confirmed to bind to HLA-A2.1 gene transfected L cells. These data suggested that peptides potentially recognized by cytotoxic T cells can bind to HLA class I molecules on tumor cells in vivo. PMID- 7493915 TI - Microsatellite instability and other molecular abnormalities in human prostate cancer. AB - Microsatellites are highly polymorphic, short-tandem repeat sequences dispersed throughout the genome. Instability of these repeat sequences at multiple genetic loci may result from mismatch repair errors, and occurs in hereditary nonpolyposis colorectal carcinoma and certain sporadic cancers. To examine microsatellite instability during the pathogenesis of human prostate cancer, we screened 48 prostate cancer cases (20 stage B, 10 stage C and 18 endocrine therapy-resistant cancer-death cases) for replication error at 17 microsatellite marker loci on 9 chromosomes. Microsatellite instabilities were found in 7 of 48 cases (14.6%), and all 7 cases showing the instability were poorly differentiated adenocarcinomas. Moreover, microsatellite instabilities were more frequently observed in cancer-death cases (6/18, 33%) than in stage B + C cases (1/30, 3.3%). These data suggest that microsatellite instability is an important genetic change related to the progression of a subset of human prostate cancer cases. It is suggested to be associated with extensive, concurrent molecular changes including androgen receptor gene mutations, as well as frequent loss of heterozygosity at chromosomal regions 8p, 10q, and 16q. PMID- 7493917 TI - Multidrug resistance-associated protein-mediated multidrug resistance modulated by cyclosporin A in a human bladder cancer cell line. AB - A doxorubicin-resistant subline (5637/DR5.5) from human bladder cancer cells (5637) was induced by stepwise increase in the doxorubicin concentration. 5637/DR5.5 cells were cross-resistant to vinblastine and etoposide but not to mitomycin C and cisplatin. We analyzed the mdr1, MRP (multidrug resistance associated protein), and DNA topoisomerase II gene expression using the reverse transcription polymerase chain reaction assay (RT-PCR) and investigated possible differences in the accumulation and efflux of radiolabeled daunorubicin. 5637/DR5.5 cells do not express the mdr1 gene, but the expression levels of MRP are markedly higher than in drug-sensitive 5637 cells. The intracellular accumulation of radiolabeled daunorubicin was markedly decreased in the 5637/DR5.5 cells in comparison with the parent cells. This reduced drug accumulation was associated with an enhanced drug efflux, but was reversed when cells were incubated with cyclosporin A. Cyclosporin A at the concentration of 5 microM caused 3.4-fold enhancement of daunorubicin-sensitivity in the 5637/DR5.5 cells. On the other hand, there was no difference in DNA-topoisomerase II activity between the parent and resistant cells. The resistance of the 5637/DR5.5 cells is therefore associated with an enhanced drug efflux mediated by the MRP gene overexpression, as distinct from P-glycoprotein, and is modulated by cyclosporin A. PMID- 7493918 TI - Protective effects of kampo medicines and baicalin against intestinal toxicity of a new anticancer camptothecin derivative, irinotecan hydrochloride (CPT-11), in rats. AB - In clinical use, irinotecan hydrochloride (CPT-11; 7-ethyl-10-[4-(piperidino)-1 piperidino]carbonyloxycamptothecin), a novel antitumor agent, causes a relatively high incidence of severe forms of diarrhea. We investigated whether baicalin, an inhibitor of beta-glucuronidase, which deconjugates the glucuronide of the active metabolite of CPT-11, SN-38 (7-ethyl-10-hydorxycamptothecin), and Japanese herbal medicines (Kampo medicines) which contain baicalin can ameliorate CPT-11-induced intestinal toxicity in rats. CPT-11 (60 mg/kg i.v. once daily for 4 consecutive days) induced intestinal toxicity characterized by diarrhea, loss of body weight, anorexia and disruption of intestinal epithelium. Treatment with baicalin (25 mg/kg p.o. twice daily) or Kampo medicines (TJ-14 and TJ-114; 1 g/kg p.o. twice daily) from the day before to 4 or 10 days after the start of CPT-11 administration resulted in significantly decreased weight loss, improved anorexia and delayed onset of diarrheal symptoms. Histological examination revealed that Kampo medicine-treated animals had less damage to the intestinal epithelium and that damage was repaired more rapidly than in control rats. These results suggest that the prophylactic use of Kampo medicines (TJ-14 and TJ-114) may be of value against CPT-11-induced intestinal toxicity. PMID- 7493920 TI - The cysteine residue responsible for the release of fibroblast growth factor-1 residues in a domain independent of the domain for phosphatidylserine binding. AB - Fibroblast growth factor (FGF)-1 lacks a classical signal sequence to direct its secretion yet utilizes high affinity cell surface receptors to signal its heparin dependent angiogenic and neurotrophic activities. We have previously reported that FGF-1 is released in response to temperature stress as a latent homodimer through a pathway that is potentiated by the Golgi inhibitor, brefeldin A (Jackson, A., Tarantini, F., Gamble, S., Friedman, S., and Maciag, T. (1995) J. Biol. Chem. 270, 33-36). In an attempt to further characterize this unconventional secretion mechanism, we sought to define the Cys residue(s) critical for FGF-1 dimer formation and release and to determine whether FGF-1 can associate with known phospholipid components of organelle or plasma membranes, which may be disturbed by brefeldin A. Utilizing FGF-1 Cys mutants, we were able to demonstrate that residue Cys30 is critical for FGF-1 release in response to heat shock. In addition, using solid phase phospholipid binding assays we demonstrate that FGF-1 is able to specifically associate with phosphatidylserine (PS). Heparin inhibits the association between FGF-1 and PS, and synthetic peptide competition assays suggest that the PS-binding domain of FGF-1 lies between residues 114 and 137. These observations indicate that FGF-1 may be able to associate with the PS component of organelle and/or plasma membranes and that the domains responsible for FGF-1 homodimer formation and PS binding are structurally distinct. PMID- 7493919 TI - Inhibition of UDP-glucuronosyltransferase by aglycons of natural glucuronides in kampo medicines using SN-38 as a substrate. AB - 7-Ethyl-10-[4-(piperidino)-1-piperidino]carbonyloxycamptothecin (CPT-11), a potent anticancer agent for lung and gynecological cancers, is metabolized in vivo to the active compound, 7-ethyl-10-hydroxycamptothecin (SN-38), which is subsequently conjugated to SN-38-glucuronide by UDP-glucuronosyltransferase (UDP GT). Three purified aglycons of natural glucuronides, baicalein, luteolin and glycyrrhetic acid, inhibited UDP-GT activity towards SN-38 as a substrate. The inhibitory potencies of these aglycons toward UDP-GT were similar to that of 1 naphthol. Based on these results, together with our previous finding that the corresponding glucuronides used in the present study strongly inhibited beta glucuronidase in gut flora, we propose that materials in Kampo (Japanese herbal) medicines containing these aglycons of natural glucuronides could be used in vivo to decrease the enterohepatic circulation of SN-38 and other drugs. PMID- 7493921 TI - Human mitogen-activated protein kinase CSBP1, but not CSBP2, complements a hog1 deletion in yeast. AB - CSBP1 and CSBP2 are human homologues of the Saccharomyces cerevisiae Hog1 mitogen activated protein kinase which is required for growth in high osmolarity media. Expression of CSBP1, but not CSBP2, complemented a hog1 delta phenotype. A CSBP2 mutant (A34V) that complements hog1 delta was isolated and found to have approximately 3-fold lower kinase activity than the wild-type CSBP2. Further analysis revealed that both the kinase activity and tyrosine phosphorylation of CSBP1 and CSBP2 (A34V) is regulated by salt. In contrast, wild-type CSBP2 is constitutively active but dependent on the upstream kinase, Pbs2. Mutagenesis studies showed that reduction or elimination of CSBP2 kinase activity restores salt responsiveness as measured by tyrosine phosphorylation suggesting that too high a level of kinase activity can result in desensitization of the host cell and inability to grow in high salt. PMID- 7493922 TI - A peptide model of basement membrane collagen alpha 1 (IV) 531-543 binds the alpha 3 beta 1 integrin. AB - Tumor cell adhesion to the triple-helical domain of basement membrane (type IV) collagen occurs at several different regions. Cellular recognition of the sequence spanning alpha 1(IV)531-543 has been proposed to be independent of triple-helical conformation (Miles, A. J., Skubitz, A. P. N., Furcht, L. T., and Fields, G. B. (1994) J. Biol. Chem. 269, 30939-30945). In the present study, integrin interactions with a peptide analog of the alpha 1(IV)-531-543 sequence have been analyzed. Tumor cell adhesion (melanoma, ovarian carcinoma) to the alpha 1(IV)531-543 chemically synthesized peptide was inhibited by a monoclonal antibody against the alpha 3 integrin subunit, and to a lesser extent by monoclonal antibodies against the beta 1 and alpha 2 integrin subunits. An anti alpha 5 monoclonal antibody and normal mouse IgG were ineffective as inhibitors of tumor cell adhesion to the peptide. Two cell surface proteins of 120 and 150 kDa bound to an alpha 1(IV)531-543 peptide affinity column and were eluted with 20 mM EDTA. When the eluted proteins were incubated with monoclonal antibodies against either the alpha 3 or beta 1 integrin subunit, proteins corresponding in molecular weight to alpha 3 and beta 1 integrin subunits were precipitated. No proteins were immunoprecipated with monoclonal antibodies against the alpha 2 or alpha 5 integrin subunits. Thus, the alpha 3 beta 1 integrin from two tumor cell types has been shown to bind directly to the alpha 1 (IV)531-543 peptide. The alpha 1(IV)531-543 peptide is the first collagen-like sequence that has been shown to bind the alpha 3 beta 1 integrin. PMID- 7493923 TI - A novel serine/threonine kinase binding the Ras-related RhoA GTPase which translocates the kinase to peripheral membranes. AB - We previously reported the cloning of a serine/threonine kinase, PAK (for p21 (Cdc42/Rac)-activated kinase), which binds to the Ras-related GTPases Cdc42Hs and Rac1 (Manser, E., Leung, T., Salihuddin, H., Zhao, Z-s., and Lim, L. (1994) Nature 367, 40-46). These p21 proteins together with RhoA comprise the Rho subfamily of proteins that are involved in morphological events. We now report the isolation of a rat cDNA encoding a 150-kDa protein, which specifically binds RhoA in its GTP form and contains an N-terminal serine/threonine kinase domain highly related to the human myotonic dystrophy kinase and a cysteine-rich domain toward the C terminus. The RhoA binding domain is unrelated to other p21 binding domains. Antibody raised against the kinase domain of the predicted protein, termed ROK alpha (for ROK alpha, RhoA-binding kinase), recognized a ubiquitous 150-kDa protein. The brain p150 purified by affinity chromatography with RhoA exhibited serine/threonine kinase activity. In cultured cells, immunoreactive p150 was recruited to membranes upon transfection with dominant positive RhoAV14 mutant and was localized with actin microfilaments at the cell periphery. These results are consistent with a role for the kinase ROK alpha as an effector for RhoA. PMID- 7493924 TI - Evidence that aspartic acid 301 is a critical substrate-contact residue in the active site of cytochrome P450 2D6. AB - Model building studies have intimated a role for aspartic acid 301 in the substrate binding of cytochrome P450 2D6 (CYP2D6). We have tested this hypothesis by generating a range of CYP2D6 mutants substituting a variety of amino acids at this site. The mutant proteins, which included substitution with a negatively charged glutamic acid residue or neutral asparagine, alanine, or glycine residues, were expressed in Saccharomyces cerevisiae. In addition, a mutant where aspartic acid 301 was deleted was also tested. All the mutants expressed approximately equivalent amounts of recombinant apoprotein and, apart from the alanine 301 and the aspartic acid 301 deletion mutants, gave carbon monoxide difference spectra of similar magnitude to the wild type. In the cases of the alanine and deletion mutants, the amount of holoprotein was significantly reduced or absent relative to the amount of apoprotein, indicating restricted heme incorporation. The glutamic acid mutant was shown to have similar catalytic properties to the wild type enzyme toward the substrates debrisoquine and metoprolol; however, some differences in regioselectivity and ligand binding were observed. The mutants containing neutral amino acids at position 301 exhibited marked reductions in catalytic activity. At low substrate concentrations little, if any, activity toward debrisoquine and metoprolol was measured. However, at a higher substrate concentration (2 mM) some activity was observed (about 10-20% of wild type levels). Consistent with the above findings, the debrisoquine-induced spin changes in the mutant proteins were markedly reduced. These data collectively demonstrate that aspartic acid 301 plays an important role in determining the substrate specificity and activity of CYP2D6 and provide experimental evidence supporting the role of this amino acid in forming an electrostatic interaction between the basic nitrogen atom in CYP2D6 substrates and the carboxylate group of aspartic acid 301. PMID- 7493925 TI - G beta gamma binds directly to the G protein-gated K+ channel, IKACh. AB - The cardiac G protein-gated K+ channel, IKACh, is activated by application of purified and recombinant beta and gamma subunits (G beta gamma) of heterotrimeric G proteins to excised inside-out patches from atrial membranes (Logothetis, D. E., Kurachi, Y., Galper, J., Neer, E., and Clapham, D. E. (1987) Nature 325, 321 326; Wickman, K., Iniguez-Lluhi, J., Davenport, P., Taussig, R. A., Krapivinsky, G. B., Linder, M. E., Gilman, A., and Clapham, D.E. (1994) Nature 368, 255-257). Cardiac IKACh is composed of two inward rectifier K+ channel subunits, GIRK1 and CIR (Krapivinsky, G., Gordon, E., Wickman, K., Velimirovic, B., Krapivinsky, L., and Clapham, D. E. (1995) Nature 374, 135-141). We show that G beta gamma directly binds to immunoprecipitated cardiac IKACh as well as to recombinant CIR and GIRK1 subunits, with dissociation constants (Kd) of 55, 50, and 125 nM, respectively. In each case, binding appeared specific as judged by competition of unlabeled G beta gamma with radiolabeled G beta gamma and inhibition of binding by antigenic peptide or G alpha-GDP, but not G alpha-GTP gamma S (guanosine 5'-3 O-(thio)triphosphate). In contrast, G alpha (GTP gamma S- or GDP-bound) did not bind to the native channel. We conclude that G beta gamma binds directly and specifically to IKACh via interactions with both CIR and GIRK1 subunits to gate the channel. PMID- 7493926 TI - Molecular cloning and characterization of AqpZ, a water channel from Escherichia coli. AB - The aquaporin family of molecular water channels is widely expressed throughout the plant and animal kingdoms. No bacterial aquaporins are known; however, sequence-related bacterial genes have been identified that encode glycerol facilitators (glpF). By homology cloning, a novel aquaporin-related DNA (aqpZ) was identified that contained no surface N-glycosylation consensus. The aqpZ RNA was not identified in mammalian mRNA by Northern analysis and exhibited bacterial codon usage preferences. Southern analysis failed to demonstrate aqpZ in mammalian genomic DNA, whereas a strongly reactive DNA was present in chromosomal DNA from Escherichia coli and other bacterial species and did not correspond to glpF. The aqpZ DNA isolated from E. coli contained a 693-base pair open reading frame encoding a polypeptide 28-38% identical to known aquaporins. When compared with other aquaporins, aqpZ encodes a 10-residue insert preceding exofacial loop C, truncated NH2 and COOH termini, and no cysteines at known mercury-sensitive sites. Expression of aqpZ cRNA conferred Xenopus oocytes with a 15-fold increase in osmotic water permeability, which was maximal after 5 days of expression, was not inhibited with HgCl2, exhibited a low activation energy (Ea = 3.8 kcal/mol), and failed to transport nonionic solutes such as urea and glycerol. In contrast, oocytes expressing glpF transported glycerol but exhibited limited osmotic water permeability. Phylogenetic comparison of aquaporins and homologs revealed a large separation between aqpZ and glpF, consistent with an ancient gene divergence. PMID- 7493927 TI - A 13-Mer peptide of a brain injury-derived protein supports neuronal survival and rescues neurons from injury caused by glutamate. AB - Neuronal survival is mediated by several kinds of proteins. Among these, neurotrophic factors play important roles in the nervous system by supporting neuronal activity and survival. It has been suggested recently that certain factors promote neuronal survival in the case of brain injury. To examine this possibility, we purified a novel neurotrophic factor from Gelfoam that was implanted at the site of injury caused in neonatal rats. During amino acid sequence analysis, we found that a fragmental peptide of this neurotrophic protein consisting of 13 amino acids showed neurotrophic activity. This 13-mer peptide promoted survival of septal cholinergic and mesencephalic dopaminergic neurons in culture and rescued hippocampal neurons from injury caused by glutamate in culture. This peptide rescued neurons from cell death caused by glutamate, even when added 4.5 h after glutamate exposure. PMID- 7493928 TI - A conserved binding motif defines numerous candidate target proteins for both Cdc42 and Rac GTPases. AB - Rho, Rac, and Cdc42 are small GTPases that regulate the formation of a variety of actin structures and the assembly of associated integrin complexes, but little is known about the target proteins that mediate their effects. Here we have used a motif-based search method to identify putative effector proteins for Rac and Cdc42. A search of the GenBankTM data base for similarity with the minimum Cdc42/Rac interactive binding (CRIB) region of a potential effector protein p65PAK has identified over 25 proteins containing a similar motif from a range of different species. These candidate Cdc42/Rac-binding proteins include family members of the mixed lineage kinases (MLK), a novel tyrosine kinase from Drosophila melanogaster (DPR2), a human protein MSE55, and several novel yeast and Caenorhabditis elegans proteins. Two murine p65PAK isoforms and a candidate protein from C. elegans, F09F7.5, interact strongly with the GTP form of both Cdc42 and Rac, but not Rho in a filter binding assay. Three additional candidate proteins, DPR2, MSE55, and MLK3 showed binding to the GTP form of Cdc42 and weaker binding with Rac, and again no interaction with Rho. These results indicate that proteins containing the CRIB motif bind to Cdc42 and/or Rac in a GTP-dependent manner, and they may, therefore, participate in downstream signaling. PMID- 7493929 TI - A novel phosphatidylserine-binding peptide motif defined by an anti-idiotypic monoclonal antibody. Localization of phosphatidylserine-specific binding sites on protein kinase C and phosphatidylserine decarboxylase. AB - A monoclonal anti-idiotypic antibody, Id8F7, previously shown to bind to a phosphatidylserine (PS)-specific binding site on protein kinase C (PKC) has been used to identify a 12-amino acid consensus sequence shared by PKC and phosphatidylserine decarboxylase (PSD). The 14-amino acid synthetic peptide derived from the corresponding region of PSD (amino acids 351-364 of the enzyme from Chinese hamster ovary cells) bound effectively and specifically to PS, and that derived from rat PKC gamma (amino acids 227-240) bound weakly but specifically to PS. Analysis of binding of Id8F7 to various synthetic peptides revealed that the consensus sequence motif, FXFXLKXXXKXR, is responsible for the interaction with both Id8F7 and PS. The results suggest that the conserved amino acid residues represent a basic structural motif for the specific interaction with PS, and the corresponding regions of PKC and PSD form the PS-specific binding sites of these enzymes. PMID- 7493930 TI - "Peptide walking" is a novel method for mapping functional domains in proteins. Its application to the Rac1-dependent activation of NADPH oxidase. AB - Activation of the superoxide generating NADPH oxidase of phagocytes involves the assembly of a multimolecular complex and is dependent on the participation of the small molecular weight GTP-binding protein Rac (1 or 2). This model system was used for mapping functional domains in the primary sequence of Rac1, based on assessing the inhibitory effect of 90 individual overlapping pentadecapeptides, spanning the entire length of Rac1, on NADPH oxidase activation in two types of cell-free assay. Five functional domains were identified, each consisting of a cluster of contiguous residues shared by members of five groups of overlapping inhibitory peptides. Four of the five domains are exposed on the molecular surface of Rac1 and were not identified previously by mutational analysis; the fifth corresponds to a polybasic motif near the carboxyl terminus, confirming earlier reports. Screening the entire linear sequence of a protein with a battery of overlapping peptides for interference with its ability to interact with upstream or downstream molecules should be of wide applicability as a reliable, fast, and economical method for mapping of functionally relevant domains. PMID- 7493931 TI - Hypoxia and mitochondrial inhibitors regulate expression of glucose transporter-1 via distinct Cis-acting sequences. AB - Studies of gene regulation by oxygen have recently defined the existence of a widely operative system that responds to hypoxia but not mitochondrial inhibitors and involves the induction of a DNA-binding complex termed hypoxia-inducible factor 1. This system has been implicated in the regulation of erythropoietin, certain angiogenic growth factors, and particular glycolytic isoenzymes. The glucose transporter Glut-1 is induced by both hypoxia and mitochondrial inhibitors, implying the operation of a different mechanism of oxygen sensing. To explore that possibility, we analyzed the cisacting sequences that convey these responses. An enhancer lying 5' to the mouse Glut-1 gene was found to convey responses both to hypoxia and to the mitochondrial inhibitors, azide and rotenone. However, detailed analysis of this enhancer demonstrated that distinct elements responded to hypoxia and the mitochondrial inhibitors. The response to hypoxia was mediated by sequences that contained a functionally critical, although atypical, hypoxia-inducible factor 1 binding site, whereas sequences lying approximately 100 nucleotides 5' to this site, which contained a critical serum response element, conveyed responses to the mitochondrial inhibitors. Thus, rather than reflecting an entirely different mechanism of oxygen sensing, regulation of Glut-1 gene expression by hypoxia and mitochondrial inhibitors arises from the function of two different sensing systems. One of these responds to hypoxia alone and resembles that involved in erythropoietin regulation, while the other responds to mitochondrial inhibitors and involves activation of a serum response element. PMID- 7493933 TI - Pleiotropic regulation of central carbohydrate metabolism in Escherichia coli via the gene csrA. AB - The carbon storage regulator gene csrA has been shown previously to dramatically affect the biosynthesis of intracellular glycogen in Escherichia coli through its negative control of the expression of two glycogen biosynthetic operons and the gluconeogenic gene pckA (Romeo, T., Gong, M., Liu, M. Y., and Brun-Zinkernagel, A. M. (1993) J. Bacteriol. 175, 4744-4755). Examination of the effects of csrA on several enzymes, genes, and metabolites of central carbohydrate metabolism now establishes a more extensive role for csrA in directing intracellular carbon flux. Phosphoglucomutase and the gluconeogenic enzymes fructose-1,6 bisphosphatase and phosphoenolpyruvate synthetase were found to be under the negative control of csrA, and these enzyme activities were maximal during the early stationary phase of growth. The enzymes glucose-6-phosphate isomerase, triose-phosphate isomerase, and enolase were positively regulated by csrA. Thus, csrA exerts reciprocal effects on glycolysis versus gluconeogenesis and glycogen biosynthesis. The glycolytic isozymes pyruvate kinase F and A (encoded by pykF and pykA, respectively) and phosphofructokinase I and II (pfkA and pfkB, respectively) exhibited differential regulation via csrA. Since the individual members of these isozyme pairs are allosterically regulated by different cellular metabolites, csrA is also capable of fine-tuning the allosteric regulation of glycolysis. In contrast, the expression of genes of the pentose phosphate pathway was weakly or negligibly affected by csrA. PMID- 7493932 TI - Expression of a novel myosin light chain kinase in embryonic tissues and cultured cells. AB - A novel, 208-kDa myosin light chain kinase (MLCK) distinct from smooth muscle and non-muscle MLCK has been identified by cross-reaction to two antibodies raised against smooth muscle MLCK. Additional antibodies directed against the amino and carboxyl termini of the smooth muscle MLCK do not react with the 208-kDa MLCK, suggesting these regions are distinct. 208-kDa MLCK phosphorylates 20-kDa myosin light chains in a Ca2+/calmodulin-dependent manner, consistent with it being a member of the MLCK family. Expression of 208-kDa MLCK and smooth muscle MLCK appears to be inversely regulated, with 208-kDa MLCK being most abundant during early development and declining at birth. In contrast, expression of smooth muscle MLCK is relatively low early during development and increases to become the predominant MLCK detected in all adult smooth and non-muscle tissues. The developmental expression pattern of the 208-kDa MLCK suggests this form be named, embryonic MLCK. PMID- 7493935 TI - Identification of an active site arginine in rat choline acetyltransferase by alanine scanning mutagenesis. AB - Kinetic as well as chemical modification studies have implicated the presence of an active site arginine in choline acetyltransferase, whose function is to stabilize coenzyme binding by interacting with the 3'-phosphate of the coenzyme A substrate. In order to identify this residue seven conserved arginines in rat choline acetyltransferase were converted to alanine by site-directed mutagenesis, and the properties of these mutants were compared with the wild type enzyme. Substitution of arginine 452 with alanine resulted in a 7-12-fold increase in the Km for both CoA and acetylcholine as well as kcat, with little change in the Km for dephospho-CoA. Product inhibition studies showed choline to be a competitive inhibitor with respect to acetylcholine, indicating R452A follows the same Theorell-Chance kinetic mechanism as the wild type enzyme. Similar results were obtained with R452Q and R452E, with the latter showing the largest changes in kinetic parameters. These findings are consistent with Arg-452 mutations increasing the rate constant, k5, for dissociation of the coenzyme from the enzyme. Direct evidence that arginine 452 is involved in coenzyme A binding was obtained by showing a 5-10-fold decrease in affinity of the R452A mutant for coenzyme A as determined by the ability to protect against phenylglyoxal inactivation as well as thermal inactivation. PMID- 7493934 TI - A rice HAL2-like gene encodes a Ca(2+)-sensitive 3'(2'),5'-diphosphonucleoside 3'(2')-phosphohydrolase and complements yeast met22 and Escherichia coli cysQ mutations. AB - A plant homolog of yeast HAL2 gene (RHL) was cloned from rice (Orizya sativa L.). The RHL cDNA complemented an Escherichia coli cysteine auxotrophic mutant, cysQ, and the yeast HAL2 mutant, met22. The latter is a methionine auxotroph and cannot use sulfate, sulfite, or sulfide as sulfur sources but exhibits wild-type activities of the enzymes necessary to assimilate sulfate and has normal sulfur uptake system. These results demonstrated that HAL2, cysQ, and RHL genes encode proteins with similar function in sulfur assimilatory pathway. The RHL cDNA expressed a 40-kDa protein that was shown to catalyze the conversion of adenosine 3'-phosphate 5'-phosphosulfate (PAPS) to adenosine 5'-phosphosulfate (APS) and 3'(2')-phosphoadenosine 5'-phosphate (PAP) to AMP. The enzyme activity is Mg(2+) dependent, sensitive to Ca2+, Li+, and Na+ and activated by K+. The inhibition by Ca2+ depends on the Mg2+/Ca2+ ratio and is reversible by high Mg2+ concentration. The substrate specificity and kinetics of RHL enzyme are very similar to the Chlorella 3'(2'),5'-diphosphonucleoside 3'(2')-phosphohydrolase (DPNPase). Our evidence suggests that this enzyme regulates the flux of sulfur in the sulfur activation pathway by converting PAPS to APS. Several residues that are essential for the activity of this enzyme were identified by site-directed mutagenesis, and the possible role of DPNPase in salt tolerance is discussed. PMID- 7493936 TI - The calmodulin-nitric oxide synthase interaction. Critical role of the calmodulin latch domain in enzyme activation. AB - The neuronal isoform of nitric oxide synthase (nNOS) requires calmodulin for nitric oxide producing activity. Calmodulin functions as a molecular switch, allowing electron transport from the carboxyl-terminal reductase domain of nitric oxide synthase to its heme-containing amino-terminal domain. Available evidence suggests that calmodulin binds to a site between the two domains of nNOS, but it is not known how calmodulin then executes its switch function. To study the calmodulin-nNOS interaction, we created a series of chimeras between calmodulin and cardiac troponin C (cTnC, a homologue of calmodulin that does not activate nNOS). Although a few chimeras showed good ability to activate nNOS, most failed to activate. A subset of the inactive chimeras retained the ability to bind to nNOS and therefore functioned as potent competitive inhibitors of nNOS activation by calmodulin (CaM). The observed inhibition was additive with the arginine antagonists NG-monomethyl-L-arginine and 7-nitroindazole, indicating a distinct and independent mechanism of nNOS inhibition. To localize the calmodulin residues that account for impaired activation in the inhibitory CaM-cTnC chimeras, we conducted a detailed mutagenesis study, replacing CaM subdomains and individual amino acid residues with the corresponding residues from cTnC. This revealed that mutations in CaM helices 2 and 6 (its latch domain) have a disproportionate negative effect on nNOS activation. Thus, our evidence suggests that the CaM latch domain plays a critical role in its molecular switch function. PMID- 7493937 TI - Cloning of a human receptor of the NPY receptor family with high affinity for pancreatic polypeptide and peptide YY. AB - Neuropeptide Y (NPY), peptide YY (PYY), and pancreatic polypeptide (PP) are structurally related peptides found in all higher vertebrates. NPY is expressed exclusively in neurons, whereas PYY and PP are produced primarily in gut endocrine cells. Several receptor subtypes have been identified pharmacologically, but only the NPY/PYY receptor of subtype Y1 has been cloned. This is a heptahelix receptor that couples to G proteins. We utilized Y1 sequence information from several species to clone a novel human receptor with 43% amino acid sequence identity to human Y1 and 53% identity in the transmembrane regions. The novel receptor displays a pharmacological profile that distinguishes it from all previously described NPY family receptors. It binds PP with an affinity (Ki) of 13.8 pM, PYY with 1.44 nM, and NPY with 9.9 nM. Because these data may identify the receptor as primarily a PP receptor, we have named it PP1. In stably transfected Chinese hamster ovary cells the PP1 receptor inhibits forskolin stimulated cAMP synthesis. Northern hybridization detected mRNA in colon, small intestine, pancreas, and prostate. As all three peptides are present in the gut through either endocrine release or innervation, all three peptides may be physiological ligands to the novel NPY family receptor PP1. PMID- 7493938 TI - Fourier transform infrared and hydrogen/deuterium exchange reveal an exchange resistant core of alpha-helical peptide hydrogens in the nicotinic acetylcholine receptor. AB - The structure of the nicotinic acetylcholine receptor (nAChR) has been studied using a novel combination of hydrogen/deuterium exchange and attenuated total reflectance Fourier transform infrared spectroscopy. Fourier transform infrared spectra show marked changes in both the amide I and amide II bands upon exposure of the nAChR to 2H2O. The substantial decrease in intensity of the amide II band reflects the exchange of roughly 30% of the peptide hydrogens within seconds of exposure to 2H2O, 50% after 30 min, 60% after 12 h, and 75% after prolonged exposure for several days at room temperature or lower temperatures. The 30% of peptide hydrogens that exchange within seconds is highly exposed to solvent and likely involved in random and turn conformations, whereas the 25% of exchange resistant peptide hydrogens is relatively inaccessible to solvent and likely located in the transmembrane domains of the nAChR. Marked changes occur in the amide I contour within seconds of exposure of the nAChR to 2H2O as a result of relatively large downshifts in the frequencies of amide I component bands assigned to turns and random structures. In contrast, only subtle change occur in the amide I contour between 3 min and 12 h after exposure to 2H2O as a result of slight downshifts in the frequencies of alpha-helix and beta-sheet vibrations. It is demonstrated that the time courses and relative magnitudes of the amide I component band shifts can be used both as an aid in the assignment of component bands to specific secondary structures and as a probe of the exchange rates of different types of secondary structures in the nAChR. Significantly, the intensities of the band shifts reflecting the exchange of alpha-helical secondary structures are relatively weak indicating that a large proportion of the 25% exchange resistant peptides adopt an alpha-helical conformation. Conversely, no evidence is found for the existence of a large number of exchange-resistant beta strands. The exchange kinetics suggest a predominantly alpha-helical secondary structure for the transmembrane domains of the nAChR. PMID- 7493939 TI - The contrahelicase activities of the replication terminator proteins of Escherichia coli and Bacillus subtilis are helicase-specific and impede both helicase translocation and authentic DNA unwinding. AB - Replication forks are arrested at sequence-specific replication termini primarily, perhaps exclusively, by polar arrest of helicase-catalyzed DNA unwinding by the terminator protein. The mechanism of this arrest is of considerable interest. This paper presents experimental evidence in support of four major points pertaining to termination of DNA replication. First, the replication terminator proteins of both Escherichia coli and Bacillus subtilis are helicase-specific contrahelicases, i.e. the proteins specifically impede the activities of helicases that are involved in symmetric DNA replication but not of those involved in conjugative DNA transfer and rolling circle replication. Second, the terminator protein (Ter) of E. coli blocks not only helicase translocation but also authentic DNA unwinding. Third, the replication terminator protein of Gram-positive B. subtilis is a polar contrahelicase of the primosomal helicase PriA of Gram-negative E. coli. Finally, the blockage of PriA-catalyzed DNA unwinding was abrogated by the passage of an RNA transcript through the replication terminator protein-terminus complex. These results are significant because of their relevance to the mechanistic aspects of replication termination. PMID- 7493940 TI - CRKL links p210BCR/ABL with paxillin in chronic myelogenous leukemia cells. AB - The Philadelphia chromosome translocation generates a chimeric oncogene, BCR/ABL, which causes chronic myelogenous leukemia (CML). In primary neutrophils from patients with CML, the major novel tyrosine-phosphorylated protein is CRKL, an SH2-SH3-SH3 linker protein which has an overall homology of 60% to CRK, the human homologue of the v-crk oncogene product. Anti-CRKL immunoprecipitates from CML cells, but not normal cells, were found to contain p210BCR/ABL and c-ABL. Several other phosphoproteins were also detected in anti-CRKL immunoprecipitates, one of which has been identified as paxillin, a 68-kDa focal adhesion protein which we have previously shown to be phosphorylated by p210BCR/ABL. Using GST-CRKL fusion proteins, the SH3 domains of CRKL were found to bind c-ABL and p210BCR/ABL, while the SH2 domain of CRKL bound to paxillin, suggesting that CRKL could physically link p210BCR/ABL to paxillin. Paxillin contains three tyrosines in Tyr-X-X-Pro (Y X-X-P) motifs consistent with amino acid sequences predicted to be optimal for binding to the CRKL-SH2 domain (at positions Tyr-31, Tyr-118, and Tyr-181). Each of these tyrosine residues was mutated to a phenylalanine residue, and in vitro binding assays indicated that paxillin tyrosines at positions 31 and 118, but not 181, are likely to be involved in CRKL-SH2 binding. These results suggest that the p210BCR/ABL oncogene may be physically linked to the focal adhesion associated protein paxillin in hematopoietic cells by CRKL. This interaction could contribute to the known adhesive defects of CML cells. PMID- 7493941 TI - Determining the requirements for cooperative DNA binding by Swi5p and Pho2p (Grf10p/Bas2p) at the HO promoter. AB - SW15 encodes a zinc finger DNA binding protein required for the transcription of the Saccharomyces cerevisiae HO gene, and PHO2 encodes a homeodomain DNA binding protein. In vitro biochemical studies using purified Swi5p and Pho2p proteins have demonstrated that Swi5p and Pho2p bind cooperatively to the HO promoter. In this report we investigate the regions of the Swi5p and Pho2p proteins required for cooperative DNA binding. The analysis of each protein gives a similar result: the zinc finger or homeodomain DNA binding domains are each sufficient for in vitro DNA binding, but additional regions of each protein are required for cooperative DNA binding. In vitro and in vivo experiments were conducted with promoters with altered spacing between the Pho2p and Swi5p binding sites. Mutations that increased the distance between the two binding sites had minimal effects on either in vitro cooperative DNA binding or in vivo upstream activating sequence activity. These observations suggest that the interaction domains of Swi5p and Pho2p are flexible and can tolerate an increase in distance between the two binding sites. The mechanism of the cooperative DNA binding by Swi5p and Pho2p is discussed. PMID- 7493942 TI - Mutational analysis of the substrate binding pocket of murine leukemia virus protease and comparison with human immunodeficiency virus proteases. AB - The differences in substrate specificity between Moloney murine leukemia virus protease (MuLV PR) and human immunodeficiency virus (HIV) PR were investigated by site-directed mutagenesis. Various amino acids, which are predicted to form the substrate binding site of MuLV PR, were replaced by the equivalent ones in HIV-1 and HIV-2 PRs. The expressed mutants were assayed with the substrate Val-Ser-Gln Asn-Tyr decreases Pro-Ile-Val-Gln-NH2 (decreases indicates the cleavage site) and a series of analogs containing single amino acid substitutions in positions P4(Ser) to P3'(Val). Mutations at the predicted S2/S2' subsites of MuLV PR have a strong influence on the substrate specificity of this enzyme, as observed with mutants H37D, V39I, V54I, A57I, and L92I. On the other hand, substitutions at the flap region of MuLV PR often rendered enzymes with low activity (e.g. W53I/Q55G). Three amino acids (His-37, Val-39, and Ala-57) were identified as the major determinants of the differences in substrate specificity between MuLV and HIV PRs. PMID- 7493943 TI - Differential regulation of Ca2+ release-activated Ca2+ influx by heterotrimeric G proteins. AB - The least understood aspect of the agonist-induced Ca2+ signal is the activation and regulation of the Ca2+ release-activated Ca2+ influx (CRAC) across the plasma membrane. To explore the possible role of heterotrimeric G proteins in the various regulatory mechanisms of CRAC, continuous renal epithelial cell lines stably expressing alpha 13 and the constitutively active alpha qQ209L were isolated and used to measure CRAC activity by the Mn2+ quench technique. Release of intracellular Ca2+ by agonist stimulation or thapsigargin was required for activation of CRAC in all cells. Although the size of the internal stores was similar in all cells, CRAC was 2-3-fold higher in alpha 13- and alpha qQ209L expressing cells. However, the channel was differentially regulated in the two cell types. Incubation at low [Ca2+]i, inhibition of the NOS pathway, or inhibition of tyrosine kinase inhibited CRAC activity in alpha 13 but not alpha qQ209L cells. Treatment with okadaic acid prevented inhibition of the channel by low [Ca2+]i and the protein kinase inhibitors in alpha 13 cells. These results suggest that expression of alpha qQ209L dominantly activates CRAC by stabilizing a phosphorylated state, whereas expression of alpha 13 makes CRAC activation completely dependent on phosphorylation by several kinases. G proteins may also modulate CRAC activity independently of the phosphorylation/dephosphorylation state of the pathway to increase maximal CRAC activity. Furthermore, our results suggest a general mechanism for regulation of CRAC that depends on coupling of receptors to specific G proteins. PMID- 7493944 TI - The role of the tyrosine kinase domain of the insulin-like growth factor-I receptor in intracellular signaling, cellular proliferation, and tumorigenesis. AB - Insulin and insulin-like growth factor (IGF-I) receptors are heterotetrameric proteins consisting of two alpha-and two beta-subunits and members of the transmembrane tyrosine kinase receptors. Specific ligand binding to the receptor triggers a cascade of intracellular events, which begins with autophosphorylation of several tyrosine residues of the beta-subunit of the receptor. The triple cluster in the tyrosine kinase domain of the beta-subunit is the earliest and major autophosphorylation site. Previous studies have shown that substitutions of these three tyrosines by phenylalanines of both insulin and IGF-I receptors practically abolish any activation of cellular signaling pathways. We have studied the effect of double tyrosine mutations on IGF-I induced receptor autophosphorylation, activation of Shc and IRS-1 pathways, and cell proliferation and tumorigenicity. Substitution of tyrosines 1131/1135 blocks any detectable autophosphorylation, whereas substitution of tyrosines 1131/1136 or 1135/1136 only reduces autophosphorylation levels in some clones by approximately 50%. Nevertheless, all the cells expressing IGF-I receptors with double tyrosine substitutions demonstrated markedly reduced signaling through Shc and IRS-1 pathways. In addition, they were unable to respond to IGF-I-stimulated cell growth in culture, and tumor formation in nude mice was abrogated. These data suggest that the presence of tyrosine 1131 or 1135 essential for receptor autophosphorylation, whereas the presence of each of these tyrosines is necessary for a fully functional receptor. PMID- 7493945 TI - Each domain of the N-ethylmaleimide-sensitive fusion protein contributes to its transport activity. AB - N-Ethylmaleimide-sensitive fusion protein (NSF) has been shown to be involved in numerous intracellular transport events. In an effort to understand the basic mechanism of NSF in vesicle-target membrane fusion events, we have examined the role that each of its three domains play in how NSF interacts with the SNAP.SNARE complex. Mutagenesis of the first ATP-binding domain (D1, amino acids 206-477) demonstrates that nucleotide binding by this domain is required for 20 S particle assembly. A second mutation, which permits ATP binding but not hydrolysis, yields a protein that can form 20 S particle but fails to mediate its disassembly. Similar mutations of the second ATP-binding domain (D2, amino acids 478-744) result in trimeric molecules that behave like wild type NSF. Domain rearrangement mutants were used to further probe the functional role of each domain. The amino terminal domain (N, amino acids 1-205) is absolutely required for binding of NSF to the SNAP.SNARE complex, because the truncated mutant, D1D2, is unable to form 20 S particle. When tested as an isolated recombinant protein, the N domain is not sufficient for binding to the SNAP.SNARE complex, but when adjacent to the D1 domain or in a trimeric molecule, the N domain does mediate binding to the SNAP.SNARE complex. Monomeric N-D1 and trimeric N-D2 could both participate in particle formation. Only the N-D1 mutant was able to facilitate MgATP-dependent release from the SNAP.SNARE complex. These data demonstrate that NSF binding to the SNAP.SNARE complex is mediated by the N domain and that both ATP binding and hydrolysis by the D1 domain are essential for 20 S particle dynamics. The intramolecular interactions outlined suggest a mechanism by which NSF may use ATP hydrolysis to facilitate the vesicle fusion process. PMID- 7493946 TI - Adapter function of protein-tyrosine phosphatase 1D in insulin receptor/insulin receptor substrate-1 interaction. AB - Insulin signal transduction involves the multisite docking protein insulin receptor substrate-1 (IRS-1) and a number of Src homology-2 (SH2) domain factors, including p85/p110 phosphatidylinositol 3-kinase, p110 GTPase-activating protein, and the phosphotyrosine-specific phosphatase PTP1D. In transfected baby hamster kidney cells, Rat1 fibroblasts, and normal IM9 lymphoblasts, PTP1D directly binds activated insulin receptor. This interaction is mediated by catalytic domain proximal SH2 determinants of the phosphatase and phosphotyrosine 1146 of the activated insulin receptor. While the receptor and the phosphatase do not serve as substrates for each other, their interaction promotes IRS-1 binding to the receptor, indicating that PTP1D functions as an adapter for insulin receptor and IRS-1. The formation of a multiprotein signaling complex involving the insulin receptor, PTP1D, and IRS-1 enhances cellular glucose uptake, a critical process in the physiological action of insulin. PMID- 7493947 TI - Interaction between cystic fibrosis transmembrane conductance regulator and outwardly rectified chloride channels. AB - We have previously described a protocol for the simultaneous isolation and reconstitution of a protein kinase A (PKA)-sensitive outwardly rectified chloride channel (ORCC) and the cystic fibrosis transmembrane conductance regulator (CFTR) from bovine tracheal epithelium. Immunoprecipitation of CFTR from this preparation prevented PKA activation of the ORCC, suggesting that CFTR regulated the ORCC and that this regulatory relationship was preserved throughout the purification procedure. We now report the purification of CFTR from bovine tracheal epithelia and the purification of a CFTR conduction mutant (G551D CFTR) from retrovirally transduced mouse L cells using a combination of alkali stripping, Triton-X extraction, and immunoaffinity chromatography. Immunopurified CFTR proteins were reconstituted in the absence and presence of ORCC. To test the hypothesis that only functional CFTR can support activation of ORCC by PKA and ATP, we used an inhibitory anti-CFTR505-511 peptide antibody or G551D CFTR. When anti-CFTR505-511 peptide antibodies were present prior to the addition of PKA and ATP, activation of both the ORCC and CFTR was prevented. If the antibody was added after activation of the ORCC and CFTR Cl- channels by PKA and ATP, only the CFTR Cl- channel was inhibited. When ORCC and G551D CFTR were co-incorporated into planar bilayers, only the ORCC was recorded and this channel could not be further activated by the addition of PKA and ATP. Thus, functional CFTR is required for activation of the ORCC by PKA and ATP. We also tested the hypothesis that PKA activation of ORCC was dependent on the extracellular presence of ATP. We added ATP on the presumed extracellular side of the lipid bilayer under conditions where it was not possible to activate the ORCC, i.e. in the presence of inhibitory anti-CFTR505-511 antibody or G551D CFTR. In both cases the ORCC regained PKA sensitivity. Moreover, the addition of hexokinase + glucose to the extracellular side prevented activation of the ORCCs by PKA and ATP in the presence of CFTR. These experiments confirm that both the presence of CFTR as well as the presence of ATP on the extracellular side is required for activation of the ORCC by PKA and ATP. PMID- 7493948 TI - Role of a distal enhancer containing a functional NF-kappa B-binding site in lipopolysaccharide-induced expression of a novel alpha 1-antitrypsin gene. AB - alpha 1-Antitrypsin (alpha 1-AT) is one of the major proteinase inhibitors in serum. Its primary physiological function is to inhibit neutrophil elastase activity in lung, but it also inhibits other serine proteases including trypsin, chymotrypsin, thrombin, and cathepsin. We have previously reported a novel alpha 1-AT, S-2 isoform, from rabbit that is induced up to 100-fold in the liver during acute inflammatory condition (Ray, B. K., Gao, X., and Ray, A. (1994) J. Biol. Chem. 269, 22080-22086). Here, we present evidence that the expression of this alpha 1-AT S-2 gene is also induced in lipopolysaccharide (LPS)-treated peripheral blood monocytes. From the cloned genomic DNA, we have identified a distal LPS-responsive enhancer located between -2438 and -1990 base pairs upstream of the transcription start site. In vitro DNA-binding studies demonstrated an interaction of an LPS-inducible NF-kappa B-like nuclear factor with a kappa B-element present in this enhancer region. Antibodies against p65 and p50 subunits of NF-kappa B supershifted the DNA-protein complex. A mutation of the NF-kappa B-binding element virtually abolished the LPS-responsive induction of the chimeric promoter in monocytic cells. Furthermore, overexpression of NF-kappa B induced the wild-type promoter activity. Taken together, these results demonstrated that during LPS-mediated inflammation, NF kappa B/Rel family of transcription factors play a crucial role in the transcriptional induction of the inflammation responsive alpha 1-AT gene. PMID- 7493949 TI - Plasma membrane Na+/H+ exchanger isoforms (NHE-1, -2, and -3) are differentially responsive to second messenger agonists of the protein kinase A and C pathways. AB - Na+/H+ exchanger (NHE) activity is regulated by several types of receptors directly coupled to distinct classes (i.e. Gs, Gi, Gq, and G12) of heterotrimeric (alpha beta gamma) GTP-binding proteins (G proteins), which, upon activation, modulate production of various second messengers (e.g. cAMP, cGMP, diacylglycerol, inositol trisphosphate, and Ca2+). Recently, four isoforms of the rat Na+/H+ exchanger were identified by molecular cloning. To examine their intrinsic responsiveness to G protein and second messenger stimulation, three of these isoforms, NHE-1, -2, and -3, were stably expressed in mutant Chinese hamster ovary cells devoid of endogenous NHE activity (AP-1 cells). Incubation of cells with either AIF4-, a general agonist of G proteins, or cholera toxin, a selective activator of G alpha s that stimulates adenylate cyclase, accelerated the rates of amiloride-inhibitable 22Na+ influx mediated by NHE-1 and -2, whereas they inhibited that by NHE-3. Similarly, short term treatment with phorbol 12 myristate 13-acetate, which mimics diacylglycerol activation of protein kinase C (PKC), or with agents (i.e. forskolin, 8-(4-chlorophenylthio)-cAMP, and isobutylmethylxanthine) that lead to activation of cAMP-dependent protein kinase (PKA) also stimulated transport by NHE-1 and NHE-2 but depressed that by NHE-3. The effects of phorbol 12-myristate 13-acetate were blocked by depleting cells of PKC or by inhibiting PKC using chelerythrine chloride, confirming a role for PKC in modulating NHE isoform activities. Likewise, the PKA antagonist, H-89, attenuated the effects of elevated cAMPi on NHE-1, -2, and -3, further demonstrating the regulation by PKA. Unlike cAMPi, elevation of cGMPi by treatment with dibutyryl-cGMP or 8-bromo-cGMP had no influence on NHE isoform activities, thereby excluding the possibility of a role for cGMP-dependent protein kinase in these cells. These data support the concept that the NHE isoforms are differentially responsive to agonists of the PKA and PKC pathways. PMID- 7493950 TI - Biosynthesis of Azorhizobium caulinodans Nod factors. Study of the activity of the NodABCS proteins by expression of the genes in Escherichia coli. AB - By in vitro and in vivo studies with Escherichia coli expressing different combinations of the nodABCS genes of Azorhizobium caulinodans, Nod factor intermediates were identified and their structures determined using mass spectrometry. Substrate-product relationships were studied by time course experiments, and the Nod factor biosynthetic pathway was partially resolved. E. coli strains, harboring nodA and/or nodB, did not produce Nod metabolites, whereas the strain expressing nodC produced chitooligosaccharides. Thus, the first committed step was the production of the carbohydrate backbone. Bacitracin and tunicamycin did not affect this step, suggesting that undecaprenyl pyrophosphate-linked intermediates were not involved. The second step was the deacetylation of chitooligosaccharides by NodB since the E. coli strain expressing nodBC produced chitooligosaccharides, deacetylated at the non-reducing end and since the NodC products were precursors of the NodBC products. A strain expressing nodBCS produced N-methylated oligosaccharides, whereas a strain expressing nodCS produced unmethylated oligosaccharides. Time course experiments showed that methylation occurred after deacetylation. Thus, NodS acted after NodB. The NodBCS metabolites were partially converted to lipo chitooligosaccharides when the nodABCS genes were expressed, showing that NodA was involved in the acylation and acted after NodS. PMID- 7493951 TI - In vivo coupling of insulin-like growth factor II/mannose 6-phosphate receptor to heteromeric G proteins. Distinct roles of cytoplasmic domains and signal sequestration by the receptor. AB - We examined the signaling function of the IGF-II/mannose 6-phosphate receptor (IGF-IIR) by transfecting IGF-IIR cDNAs into COS cells, where adenylyl cyclase (AC) was inhibited by transfection of constitutively activated G alpha i cDNA (G alpha i2Q205L). In cells transfected with IGF-IIR cDNA, IGF-II decreased cAMP accumulation promoted by cholera toxin or forskolin. This effect of IGF-II was not observed in untransfected cells or in cells transfected with IGF-IIRs lacking Arg2410-Lys2423. Thus, IGF-IIR, through its cytoplasmic domain, mediates the Gi linked action of IGF-II in living cells. We also found that IGF-IIR truncated with C-terminal 28 residues after Ser2424 caused G beta gamma-dominant response of AC in response to IGF-II by activating Gi. Comparison with the G alpha i dominant response of AC by intact IGF-IIR suggests that the C-terminal 28-residue region inactivates G beta gamma. This study not only provides further evidence that IGF-IIR has IGF-II-dependent signaling function to interact with heteromeric G proteins with distinct roles by different cytoplasmic domains, it also suggests that IGF-IIR can separate and sequestrate the G alpha and G beta gamma signals following Gi activation. PMID- 7493952 TI - From xenobiotic to antibiotic, formation of protoanemonin from 4-chlorocatechol by enzymes of the 3-oxoadipate pathway. AB - Chloroaromatics, a major class of industrial pollutants, may be oxidatively metabolized to chlorocatechols by soil and water microorganisms that have evolved catabolic activities toward these xenobiotics. We show here that 4-chlorocatechol can be further transformed by enzymes of the ubiquitous 3-oxoadipate pathway. However, whereas chloromuconate cycloisomerases catalyze the dechlorination of 3 chloro-cis,cis-muconate to form cis-dienelactone, muconate cycloisomerases catalyze a novel reaction, i.e. the dechlorination and concomitant decarboxylation to form 4-methylenebut-2-en-4-olide (protoanemonin), an ordinarily plant-derived antibiotic that is toxic to microorganisms. PMID- 7493953 TI - Epitope-labeled soluble human interleukin-5 (IL-5) receptors. Affinity cross-link labeling, IL-5 binding, and biological activity. AB - The human receptor for the potent eosinophilopoietic cytokine interleukin-5 (IL 5) consists of two components: a 60-kDa ligand-binding alpha chain (IL-5 alpha R) and a 130-kDa beta chain (IL-5 beta R). Three ectodomain constructs of the alpha chain (alpha RED) bearing C-terminal epitope tags were engineered and expressed in baculovirus-infected Sf9 cells. Each recombinant alpha chain was secreted into the medium, maximum expression occurring 72 h post-infection. The various soluble alpha chains were shown by affinity cross-link labeling and competition with unlabeled IL-5 to bind recombinant human (rh) 125I-IL-5 specifically with an ED50 of 2-5 nM. The epitope tag provided a simple purification of the receptor from conditioned medium using immunoaffinity chromatography. The purified material had an apparent molecular mass of 43 kDa and was heterogeneously glycosylated. Sedimentation analysis revealed a 1:1 association of the purified epitope-tagged soluble receptor with its ligand, resulting in the formation of a 70-74-kDa complex. Circular dichroism analysis revealed that the soluble alpha chain existed with a significantly ordered structure consisting of 42% beta-sheet and 6% alpha-helix. Such analyses combined with fluorescence spectrometry suggested that ligand-receptor complex formation in solution resulted in minimal conformational changes, consistent with the suggestion that the membrane associated form of the alpha chain itself has minimal signal transduction capability. Surface plasmon resonance studies of the interaction of the purified alpha RED with immobilized rhIL-5 revealed a specific, competable interaction with a dissociation constant of 9 nM. Preincubation of an IL-5-dependent cell line with the epitope-tagged alpha RED also dose-dependently neutralized rhIL-5 induced proliferation. These data demonstrate that biologically active epitope tagged recombinant soluble IL-5 receptors are facile to produce in large quantities and may have therapeutic utility in the modulation of IL-5-dependent eosinophilia in man. PMID- 7493954 TI - S-nitrosation of serum albumin by dinitrosyl-iron complex. AB - The objective of this study was to identify a potential mechanism for S nitrosation of proteins. Therefore, we assessed S-nitrosation of bovine serum albumin by dinitrosyl-iron-di-L-cysteine complex [(NO)2Fe(L-cysteine)2], a compound similar to naturally occurring iron-nitrosyls. Within 5-10 min, (NO)2Fe(L-cysteine)2 generated paramagnetic albumin-bound dinitrosyl-iron complex and S-nitrosoalbumin in a ratio of 4:1. Although S-nitroso-L-cysteine was concomitantly formed in low amounts, its concentration was not sufficient to account for formation of S-nitrosoalbumin via a trans-S-nitrosation reaction. Low oxygen tension did not affect S-nitrosation by the dinitrosyl-iron complex thus excluding the involvement of oxygenated NOx-species in the nitrosation reaction. Blockade of albumin histidine residues by pyrocarbonate, which prevented formation of dinitrosyl-iron-albumin complex, did not inhibit S-nitrosation of albumin. Thus, S-nitrosation of albumin by (NO)2Fe(L-cysteine)2 can proceed by direct attack of a nitrosyl moiety on the protein thiolate, without previous binding of the iron. We conclude that protein-bound dinitrosyl-iron complexes detected in high concentrations in certain tissues provide a reservoir of S nitrosating species, e.g. low molecular dinitrosyl iron complexes. PMID- 7493956 TI - Crystallographic studies of the interaction of cyclodextrin glycosyltransferase from Bacillus circulans strain 251 with natural substrates and products. AB - Asp-229, Glu-257, and Asp-328 constitute the catalytic residues in cyclodextrin glycosyl transferase from Bacillus circulans strain 251. Via site-directed mutagenesis constructed D229N, E257Q, and D328N mutant proteins showed a 4,000 60,000-fold reduction of cyclization activity. A D229N/E257Q double mutant showed a 700,000-fold reduction and was crystallized for use in soaking experiments with alpha-cyclodextrin. Crystal structures were determined of wild type CGTase soaked at elevated pH with alpha-cyclodextrin (resolution, 2.1 A) and maltoheptaose (2.4 A). In addition, structures at cryogenic temperature were solved of the unliganded enzyme (2.2 A) and of the D229N/E257Q mutant after soaking with alpha cyclodextrin (2.6 A). In the crystals soaked in alpha-cyclodextrin and maltoheptaose, a maltotetraose molecule is observed to bind in the active site. Residue 229 is at hydrogen bonding distance from the C-6 hydroxyl group of the sugar, which after cleavage will contain the new reducing end. In the D229N/E257Q double mutant structure, two alpha-cyclodextrins are observed to replace two maltoses at the E-domain, thus providing structural information on product inhibition via binding to the enzyme's raw starch binding domain. PMID- 7493955 TI - Reaction versus subsite stereospecificity of peptidylglycine alpha-monooxygenase and peptidylamidoglycolate lyase, the two enzymes involved in peptide amidation. AB - Carboxyl-terminal amidation, a required post-translational modification for the bioactivation of many neuropeptides, entails sequential enzymatic action by peptidylglycine alpha-monooxygenase (PAM, EC 1.14.17.3) and peptidylamidoglycolate lyase (PGL, EC 4.3.2.5). The monooxygenase, PAM, first catalyzes conversion of a glycine-extended pro-peptide to the corresponding alpha hydroxyglycine derivative, and the lyase, PGL, then catalyzes breakdown of this alpha-hydroxyglycine derivative to the amidated peptide plus glyoxylate. We have previously established that PAM and PGL exhibit tandem reaction stereospecificities, with PAM producing, and PGL being reactive toward, only alpha-hydroxyglycine derivatives of absolute configuration (S). We now demonstrate that PAM and PGL exhibit dramatically different subsite stereospecificities toward the residue at the penultimate position (the P2 residue) in both substrates and inhibitors. Incubation of Ac-L-Phe-Gly, Ac-L-Phe L-Phe-Gly, or (S)-O-Ac-mandelyl-Gly with PAM results in complete conversion of these substrates to the corresponding alpha-hydroxylated products, whereas the corresponding X-D-Phe-Gly compounds undergo conversions of < 1%. The KI of Ac-D Phe-Gly is at least 700-fold higher than that of Ac-L-Phe-Gly, and the same pattern holds for other substrate stereoisomers. This S2 subsite stereospecificity of PAM also holds for competitive inhibitors; thus, the KI of 45 microM for Ac-L-Phe-OCH2CO2H increases to 2,247 microM for the -D-Phe- enantiomer. In contrast, incubation of PGL with Ac-L-Phe-alpha-hydroxy-Gly, Ac-D Phe-alpha-hydroxy-Gly, (S)-O-Ac-mandelyl-alpha-hydroxy-Gly, or (R)-O-Ac-mandelyl alpha-hydroxy-Gly results in facile enzymatic conversion of each of these compounds to their corresponding amide products. The simultaneous expression of high reaction stereospecificity and low S2 subsite stereospecificity in the course of PGL catalysis was illustrated by a series of experiments in which enzymatic conversion of the diastereomers of Ac-L-Phe-alpha-hydroxy-Gly and Ac-D Phe-alpha-hydroxy-Gly was monitored directly by HPLC. Kinetic parameters were determined for both substrates and potent competitive inhibitors of PGL, and the results confirm that, in sharp contrast to PAM, the configuration of the chiral moiety at the P2 position has virtually no effect on binding or catalysis. These results illustrate a case where catalytic domains, which must function sequentially (and with tandem reaction stereochemistry) in a given metabolic process, nevertheless exhibit sharply contrasting subsite stereospecificities toward the binding of substrates and inhibitors. PMID- 7493958 TI - Identification of functional domains of the aryl hydrocarbon receptor. AB - Functional domains of the mouse aryl hydrocarbon receptor (Ahr) were investigated by deletion analysis. Ligand binding was localized to a region encompassing the PAS B repeat. The ligand-mediated dissociation of Ahr from the 90-kDa heat shock protein (HSP90) does not require the aryl hydrocarbon receptor nuclear translocator (Arnt), but it is slightly enhanced by this protein. One HSP90 molecule appears to bind within the PAS region. The other molecule of HSP90 appears to require interaction at two sites: one over the basic helix-loop-helix region, and the other located within the PAS region. Each mutant was analyzed for dimerization with full-length mouse Arnt and subsequent binding of the dimer to the xenobiotic responsive element (XRE). In order to minimize any artificial steric hindrances to dimerization and XRE binding, each Ahr mutant was also tested with an equivalently deleted Arnt mutant. The basic region of Ahr is required for XRE binding but not for dimerization. Both the first and second helices of the basic helix-loop-helix motif and the PAS region are required for dimerization. These last results are analogous to those previously obtained for Arnt (Reisz-Porszasz, S., Probst, M.R., Fukunaga, B. N., and Hankinson, O. (1994) Mol. Cell. Biol. 14, 6075-6086) compatible with the notion that equivalent regions of Ahr and Arnt associate with each other. Deletion of the carboxyl terminal half of Ahr does not affect dimerization or XRE binding but, in contrast to an equivalent deletion of Arnt, eliminates biological activity as assessed by an in vivo transcriptional activation assay, suggesting that this region of Ahr plays a more prominent role in transcriptional activation of the cyp1a1 gene than the corresponding region of Arnt. PMID- 7493957 TI - The spin trap alpha-(4-pyridyl-1-oxide)-N-tert-butylnitrone stimulates peroxidase mediated oxidation of deferoxamine. Implications for pharmacological use of spin trapping agents. AB - The iron chelator deferoxamine (Desferal; DSFL) reacts with peroxidases and H2O2 to form the DSFL radical (DSFL.), which can be detected by EPR spectroscopy. We have found that DSFL. formation resulting from exposure to H2O2 and any of a number of different peroxidases is greatly enhanced in the presence of the nitrone spin trap alpha-(4-pyridyl-1-oxide)-N-tert-butylnitrone (4-POBN). This enhancement was seen at 4-POBN concentrations as low as 200 microM. We observed a modest enhancement of DSFL. formation with 2-methyl-2-nitrosopropane. However, no enhancement was seen with 5,5-dimethyl-1-pyrroline 1-oxide (DMPO) or phenyl-tert butylnitrone. A modest enhancement was also seen with the nitrone compound pyridine N-oxide. 2-Methyl-2-nitrosopropane and pyridine N-oxide were additionally capable of increasing enzymatic peroxidase activity as measured by o dianisidine and/or tetramethylbenzidine oxidation. Furthermore, at high concentrations of 4-POBN (50 mM) in the absence of DSFL, we detected a peroxidase/H2O2-dependent 12-line EPR spectrum that likely represents a 4-POBN/.4 POBN nitrogen-centered spin adduct. In the presence of both 4-POBN (10 mM) and DMPO (100 mM), an 18-line EPR spectrum was observed consistent with formation of a DMPO/.4-POBN nitrogen-centered spin adduct. Thus, the nitrone spin trap 4-POBN can enhance the peroxidase-mediated formation of DSFL., possibly via the formation of a transient 4-POBN radical species. These data suggest the importance of assessing the potential for nitrone spin traps to both inhibit and enhance biological oxidation prior to their use as potential pharmacological agents. PMID- 7493959 TI - Inhibition of influenza-induced membrane fusion by lysophosphatidylcholine. AB - Lysolipids have been reported to inhibit various membrane fusion events, and it was suggested that inhibition was due to their "inverted cone" shape, which hinders the formation of intermediate lipid structures required for fusion (Chernomordik, L. V., Vogel, S. S., Sokoloff, A., Onaran, H. O., Leikina, E. A., and Zimmerberg, J. (1993) FEBS Lett. 318, 71-76). Here, the effect of lysophosphatidylcholine (LPC) on fusion mediated by the hemagglutinin (HA) of influenza virus was investigated. Virus-liposome fusion was inhibited by LPC if the lysolipid was added to the membranes from an aqueous stock solution but not if LPC was symmetrically distributed over both leaflets of the liposomal bilayer. These findings would be consistent with an effect of LPC on lipid intermediate formation, but inhibition increased with increasing acyl chain length and thus a less pronounced inverted cone shape of the lysolipids suggesting that the mechanism of inhibition might be different. At low pH, due to the exposure of the fusion peptide of HA, followed by its insertion into the liposomal membrane, virus acquires the ability to bind to zwitterionic liposomes lacking receptors for HA. This type of binding was inhibited by LPC. Moreover, leakage of calcein from receptor-containing liposomes, induced by purified HA at low pH, was inhibited by LPC. Therefore, the inhibition of influenza-induced fusion by LPC was caused by the binding of LPC to fusion peptides, thereby preventing their interaction with the target membrane rather than an effect on intermediate lipid structures. PMID- 7493961 TI - The deletion of petG in Chlamydomonas reinhardtii disrupts the cytochrome bf complex. AB - The 4-kDa protein encoded by chloroplast petG copurifies with the cytochrome bf complex of spinach and is found in a number of other photosynthetic organisms, including the eukaryotic alga Chlamydomonas reinhardtii. To determine whether petG is involved in the function or assembly of the cytochrome bf complex, the gene was cloned from C. reinhardtii, excised from the DNA fragment, and replaced with a spectinomycin resistance cassette. A petG deletion strain of C. reinhardtii was then obtained by biolistic transformation. The resulting homoplasmic petG deletion strains are unable to grow photosynthetically, and immunoblot analysis shows markedly decreased levels of cytochrome b6, cytochrome f, the Rieske iron-sulfur protein, and subunit IV. To verify that this phenotype was due to the removal of petG, we also constructed a strain with a deletion in the open reading frame (ORF56), which is found 25 base pairs downstream of petG. The ORF56 deletion strain grew photosynthetically and had wild-type levels of the four major cytochrome bf subunits. We conclude that the absence of the PetG protein affects either the assembly or stability of the cytochrome bf complex in C. reinhardtii. PMID- 7493960 TI - Activation and nuclear translocation of mitogen-activated protein kinases by polyomavirus middle-T or serum depend on phosphatidylinositol 3-kinase. AB - Several cellular signal transduction pathways activated by middle-T in polyomavirus-transformed cells are required for viral oncogenicity. Here we focus on the role of phosphatidylinositol 3-kinase (PI 3-kinase) and Ras and address the question how these signaling molecules cooperate during cell cycle activation. Ras activation is mediated through association with SHC.GRB2.SOS and leads to increased activity of several members of the mitogen-activated protein (MAP) kinase family, while activation of PI 3-kinase results in the generation of D3-phosphorylated phosphatidylinositides whose downstream targets remain elusive. PI 3-kinase activation might also ensue as a direct consequence of Ras activation. Oncogenicity of middle-T requires stimulation of both Ras- and PI 3 kinase-dependent pathways. Mutants of middle-T incapable to bind either SHC.GRB2.SOS or PI 3-kinase are not oncogenic. Sustained activation and nuclear localization of one of the MAP kinases, ERK1, was observed in wild type but not in mutant middle-T-expressing cells. Wortmannin, an inhibitor of PI 3-kinase, prevented MAP kinase activation and nuclear localization in middle-T-transformed cells. PI 3-kinase activity was also required for activation of the MAP kinase pathway in normal serum-stimulated cells, generalizing the concept that signaling through MAP kinases requires not only Ras-but also PI 3-kinase-mediated signals. PMID- 7493962 TI - Multimerization determinants reside in both the catalytic core and C terminus of avian sarcoma virus integrase. AB - We have shown previously that the active form of avian sarcoma virus integrase (ASV IN) is a multimer. In this report we investigate IN multimerization properties by a variety of methods that include size exclusion chromatography, chemical cross-linking, and protein overlay assays. We show that removal of the nonconserved C-terminal region of IN results in a reduced capacity for multimerization, whereas deletion of the first 38 amino acids has little effect on the oligomeric state. Binding of a full-length IN fusion protein to various IN fragments indicates that sequences in both the catalytic core (residues 50-207) and a C-terminal region (residues 201-240) contribute to IN self-association. We also observe that the isolated C-terminal fragment (residues 201-286) is capable of self-association. Finally, a single amino acid substitution in the core domain (S85G) produces a severe defect in multimerization. We conclude from these analyses that both the catalytic core and a region in the nonconserved C terminus are involved in ASV integrase multimerization. These results enhance our understanding of intergrase self-association determinants and define a major role of the C-terminal region of ASV integrase in this process. PMID- 7493963 TI - Identification of a novel Giardia lamblia cyst wall protein with leucine-rich repeats. Implications for secretory granule formation and protein assembly into the cyst wall. AB - Giardia lamblia trophozoites, like most intestinal parasitic protozoa, undergo fundamental biological changes to survive outside the intestine of their mammalian host by differentiating into infective cysts. This complex process entails the coordinated production, processing, and transport of cyst wall constituents for assembly into a protective cyst wall. Yet, little is known about this process and the identity of cyst wall constituents. We previously identified a 26-kDa cyst wall protein, CWP1. In the present work, using monoclonal antibodies to cyst wall antigens, we cloned the gene that encodes a novel 39-kDa cyst wall protein, CWP2. Expression of CWP1 and CWP2 was induced during encystation with identical kinetics. Soon after synthesis, these two proteins combine to form a stable complex, which is concentrated within the encystation specific secretory granules before incorporation into the cyst wall. Both proteins contain five tandem copies of a 24-residue leucine-rich repeat, a motif implicated in protein-protein interactions. Unlike CWP1, CWP2 has an extremely basic 121-residue COOH-terminal extension that might be involved in the sorting of these proteins to the secretory granules. PMID- 7493964 TI - The conserved noncatalytic 40-residue sequence in cellulases and hemicellulases from anaerobic fungi functions as a protein docking domain. AB - Two cDNAs, designated xynA and manA, encoding xylanase A (XYLA) and mannanase A (MANA), respectively, were isolated from a cDNA library derived from mRNA extracted from the anaerobic fungus, Piromyces. XYLA and MANA displayed properties typical of endo-beta 1,4-xylanases and mannanases, respectively. Neither enzyme hydrolyzed cellulosic substrates. The nucleotide sequences of xynA and manA revealed open reading frames of 1875 and 1818 base pairs, respectively, coding for proteins of M(r) 68,049 (XYLA) and 68,055 (MANA). The deduced primary structure of MANA revealed a 458-amino acid sequence that exhibited identity with Bacillus and Pseudomonas fluorescens subsp. cellulosa mannanases belonging to glycosyl hydrolase Family 26. A 40-residue reiterated sequence, which was homologous to duplicated noncatalytic domains previously observed in Neocallimastix patriciarum xylanase A and endoglucanase B, was located at the C terminus of MANA. XYLA contained two regions that exhibited sequence identity with the catalytic domains of glycosyl hydrolase Family 11 xylanases and were separated by a duplicated 40-residue sequence that exhibited strong homology to the C terminus of MANA. Analysis of truncated derivatives of MANA confirmed that the N-terminal 458-residue sequence constituted the catalytic domain, while the C terminal domain was not essential for the retention of catalytic activity. Similar deletion analysis of XYLA showed that the C-terminal catalytic domain homologue exhibited catalytic activity, but the corresponding putative N-terminal catalytic domain did not function as a xylanase. Fusion of the reiterated noncatalytic 40-residue sequence conserved in XYLA and MANA to glutathione S transferase, generated a hybrid protein that did not associate with cellulose, but bound to 97- and 116-kDa polypeptides that are components of the multienzyme cellulase-hemicellulase complexes of Piromyces and Neocallimastix patriciarum, respectively. The role of this domain in the assembly of the enzyme complex is discussed. PMID- 7493966 TI - Cloning and functional expression of a soluble form of kynurenine/alpha aminoadipate aminotransferase from rat kidney. AB - Several aminotransferases with kynurenine aminotransferase (KAT) activity are able to convert L-kynurenine into kynurenic acid, a putative endogenous modulator of glutamatergic neurotransmission. In the rat, one of the described KAT isoforms has been found to correspond to glutamine transaminase K. In addition, rat kidney alpha-aminoadipate aminotransferase (AadAT) also shows KAT activity. In this report, we describe the isolation of a cDNA clone encoding the soluble form of this aminotransferase isoenzyme from rat (KAT/AadAT). Degenerate oligonucleotides were designed from the amino acid sequences of rat kidney KAT/AadAT tryptic peptides for use as primers for reverse transcription-polymerase chain reaction of rat kidney RNA. The resulting polymerase chain reaction fragment was used to screen a rat kidney cDNA library and to isolate a cDNA clone encoding KAT/AadAT. Analysis of the combined DNA sequences indicated the presence of a single 1275 base pair open reading frame coding for a soluble protein of 425 amino acid residues. KAT/AadAT appears to be structurally homologous to aspartate aminotransferase in the pyridoxal 5'-phosphate binding domain. RNA blot analysis of rat tissues, including brain, revealed a single species of KAT/AadAT mRNA of approximately 2.1 kilobases. HEK-293 cells transfected with the KAT/AadAT cDNA exhibited both KAT and AadAT activities with enzymatic properties similar to those reported for the rat native protein. PMID- 7493967 TI - Molecular cloning, DNA sequence analysis, and biochemical characterization of a novel 65-kDa FK506-binding protein (FKBP65). AB - We have identified a mouse gene encoding a 65-kDa protein (FKBP65) that shares homology with members of the FK506-binding protein (FKBP) class of immunophilins. Predicted amino acid sequence shows that this protein shares significant homology with FKBP12 (46%), FKBP13 (43%), FKBP25 (35%), and FKBP52 (26%). FKBP65 contains four predicted peptidylprolyl cistrans-isomerase (PPIase) signature domains, and, although similar in size, is distinct from FKBP52 (also identified as FKBP59, hsp56, or HBI), which contains three FKBP12-like PPIase domains. With N-succinyl Ala-Ala-Pro-Phe-p-nitroanilide as the substrate, recombinant FKBP65 is shown to accelerate the isomerization of the prolyl peptide bond with a catalytic efficiency similar to other family members. This isomerization activity is inhibited by FK506 and rapamycin, but is not sensitive to Cyclosporin A. Based on Northern blot analysis, FKBP65 mRNA transcripts are present in lung, spleen, heart, brain, and testis. A polyclonal antibody, raised against a COOH-terminal peptide (amino acid residues 566-581), was used to immunoprecipitate FKBP65 from NIH3T3 cells and demonstrate that FKBP65 is a glycoprotein. In addition, [32P]orthophosphate labeling experiments show that FKBP65 is also a phosphoprotein. These results suggest that FKBP65 is a new FKBP family member. PMID- 7493970 TI - Structure-function relationships of the mitochondrial bc1 complex in temperature sensitive mutants of the cytochrome b gene, impaired in the catalytic center N. AB - Seven new structures of cytochrome b have been recently identified by isolating and sequencing revertants from cytochrome b respiratory deficient mutants (Coppee, J. Y., Brasseur, G., Brivet-Chevillotte, P., and Colson, A. M. (1994) J. Biol. Chem. 269, 4221-4226). These mutations are located in the center N domain (QN). All the revertants exhibited a modified heme b562 maximum, confirming that part of the NH2-terminal region is in the vicinity of the extramembranous loop between helices IV-V and heme b562. Based on measurements performed on the maximal activities occurring in each segment of the respiratory chain, the decrease observed in the NADH oxidase activities of several revertants was correlated with some bc1 complex activity impairments; this may also explain why a moderate decrease in bc1 complex activity does not limit the succinate oxidase activity. The decrease in the rate of reduction of cytochrome b via the center N pathway is responsible for the impairment of the bc1 complex activity of these revertants. The three double-mutated revertants (S206L/N208K or -Y; S206L/W30C) are temperature-sensitive in vivo, and their mitochondria like that of the original mutant S206L are thermosensitive in vitro. Isolating the W30C mutation does not yield a thermosensitive phenotype: the replacement of serine 206 by leucine is therefore responsible for the thermoinstability of these strains; this temperature sensitivity is reinforced by additional mutations N208K or N208Y, and not by W30C. These data suggest that serine 206 and asparagine 208 are involved in the thermostability of the protein. When bc1 complex activity is lost after incubating mitochondria at a nonpermissive temperature (37 degrees C), heme b is still present, but can no longer be reduced by physiological substrate. The progressive loss of bc1 complex activity seems to be initially linked to a change in the tertiary structure of cytochrome b, which occurs drastically at center N and much more slowly at center P, as shown by kinetic study on the two cytochrome b redox pathways. PMID- 7493968 TI - Purification and characterization of the cytochrome b6 f complex from Chlamydomonas reinhardtii. AB - A protocol has been developed for the purification of the cytochrome b6 f complex from the unicellular alga Chlamydomonas reinhardtii. It is based on the use of the neutral detergent Hecameg (6-O-(N-heptylcarbamoyl)-methyl-alpha-D glycopyranoside) and comprises only three steps: selective solubilization from thylakoid membranes, sucrose gradient sedimentation, and hydroxylapatite chromatography. The purified complex contains two b hemes (alpha bands, 564 nm; Em,8 = -84 and -158 mV) and one chlorophyll alpha (lambda max = 667-668 nm) per cytochrome f (alpha band, 554 nm; Em,8 = +330 mV). It is highly active in transferring electrons from decylplastoquinol to oxidized plastocyanin (turnover number 250-300 s-1). The purified complex contains seven subunits, whose identity has been established by N-terminal sequencing and/or peptide-specific immunolabeling, namely four high molecular weight subunits (cytochrome f, Rieske iron-sulfur protein, cytochrome b6, and subunit IV) and three approximately 4-kDa miniproteins (PetG, PetL, and PetX). Stoichiometry measurements are consistent with every subunit being present as two copies per b6 f dimer. PMID- 7493969 TI - Kinetic analysis of the fate of nitric oxide synthesized by macrophages in vitro. AB - To investigate the fate of nitric oxide (NO) synthesized by activated macrophages, the concentrations of NO and its principal reaction products, nitrite (NO2-) and nitrate (NO3-), were measured as a function of time in suspension cultures of RAW264.7 macrophages attached to microcarrier beads. Synthesis of NO became evident 2-5 h after stimulation of the cells, and steady concentrations of NO were achieved after about 9 h. The appearance of NO in the extracellular fluid coincided with the appearance of NO2- and NO3-, which were formed thereafter at approximately equal and constant rates. Using a kinetic model based on rate constants measured previously in cell-free systems, only half of the NO2- formed could be accounted for by the reaction of NO with O2. It is known that NO reacts with superoxide (O2.) to give peroxynitrite and that NO also reacts with peroxynitrite to yield NO2-, so that the latter reaction may explain the "excess" NO2- formation. Adding superoxide dismutase to the medium markedly reduced the ratio of NO3- to NO2-, consistent with the hypothesis that NO3- in the medium results primarily from the extracellular reaction of NO with O2-.. The addition of morpholine, a model amine, resulted in formation of N nitrosomorpholine, concurrent with the other products. Measured rates of nitrosomorpholine formation were 6-fold lower than predictions based on kinetics in simple solutions, suggesting that in the cell culture system there were additional reactions that lowered the concentration of nitrous anhydride, the principal nitrosating agent formed from NO and O2. PMID- 7493965 TI - Essential amino acids regulate fatty acid synthase expression through an uncharged transfer RNA-dependent mechanism. AB - To better understand the regulation of gene expression by amino acids, we studied the effects of these macronutrients on fatty acid synthase (FAS), an enzyme crucial for energy storage. When HepG2 cells were fed serum-free media selectively deficient in each amino acid, the omission of any single classic essential amino acid as well as Arg or His (essential in some rapidly growing cells) resulted in FAS mRNA levels that were about half of those in complete medium. Control message levels were unaffected and omission of nonessential amino acids did not alter FAS expression. FAS mRNA levels peaked 12-16 h after feeding complete and Ser (nonessential)-deficient media but did not increase in cells fed Lys (essential)-deficient medium. With Lys, FAS mRNA increased over the physiologic concentration range of 15-150 microM, and low concentrations of lysine decreased FAS but not apoB protein mass. Transcription inhibitors mimicked treatment with Lys-deficient media, and nuclear run-off assays showed that Lys deficient media abolished FAS but not apoB transcription. After treatment with Lys-deficient media, the intracellular Lys pool was rapidly depleted in association with an increase of uncharged (deacylated) tRNA Lys from < 1 to 64% of available tRNA Lys. Even in the presence of the essential amino acid His, increasing the level of uncharged tRNA His with histidinol, a competitive inhibitor of the histidinyl-tRNA synthetase, blocked FAS expression. Tyrosinol treatment did not alter FAS mRNA levels. These results suggest that essential amino acids regulate FAS expression by altering uncharged tRNA levels, a novel mechanism for nutrient control of gene expression in mammalian cells. PMID- 7493972 TI - Arginine 120 of prostaglandin G/H synthase-1 is required for the inhibition by nonsteroidal anti-inflammatory drugs containing a carboxylic acid moiety. AB - The therapeutic action of nonsteroidal anti-inflammatory drugs (NSAIDs) is exerted through the inhibition of prostaglandin G/H synthase (PGHS), which is expressed as two isoenzymes, termed PGHS-1 and PGHS-2. From the crystal structure of sheep PGHS-1, it has been proposed that the carboxylic acid group of flurbiprofen is located in a favorable position for interacting with the arginine 120 residue of PGHS-1 (Picot, D., Loll, P. J., and Garavito, R. M. (1994) Nature 367, 243-249). Mutation of this Arg120 residue to Glu was performed and expressed in COS-7 cells using a vaccinia virus expression system. Comparison of microsomal enzyme preparations show that the mutation results in a 20-fold reduction in the specific activity of PGHS-1 and in a 100-fold increase in the apparent Km for arachidonic acid. Indomethacin, flurbiprofen, and ketoprofen, inhibitors of PGHS activity containing a free carboxylic acid group, do not exhibit any inhibitory effects against the activity of PGHS-1(Arg120-->Glu). Diclofenac and meclofenamic acid, other NSAIDs containing a free carboxylic acid group, were 50-100-fold less potent inhibitors of the activity of the mutant as compared with the wild type PGHS. In contrast, the nonacid PGHS inhibitors, 5-bromo-2-(4-fluorophenyl)-3-(4 methylsulfonyl)thiophene (DuP697) and a desbromo-sulfonamide analogue of DuP697 (L-746,483), were both more potent inhibitors of PGHS-1(Arg120-->Glu) than of the wild tyupe PGHS-1. Inhibition of PGHS-1(Arg120-->Glu) was time-dependent for diclofenac and time-independent for DuP697, as observed for the wild type enzyme, indicating that the mutation does not alter the basic mechanism of inhibition. Aspirin is an acid NSAID that inhibits PGHS-1 through a unique covalent acetylation of the enzyme and also showed a reduced rate of inactivation of the mutated enzyme. These data provide biochemical evidence of the importance of the Arg120 residue in PGHS-1 for interaction with arachidonic acid and NSAIDs containing a free carboxylic acid moiety. PMID- 7493971 TI - Molecular characteristics of Na(+)-coupled glucose transporters in adult and embryonic rat kidney. AB - Two distinct Na(+)-coupled glucose transporters (SGLTs) with either a high or a low affinity for glucose were shown to provide reabsorption of filtered glucose in the kidney. We have previously reported the characteristics of the high affinity Na+/glucose cotransporter SGLT1 from rabbit, rat, and human kidney and the low affinity Na+/glucose cotransporter SGLT2 from human kidney. Because the molecular identity of SGLT2 as the kidney cortical low affinity Na+/glucose cotransporter has been recently challenged based on studies of the porcine low affinity Na+/glucoe cotransporter SAAT-pSGLT2 (Mackenzie, B., Panayotova Heiermann, M., Loo, D. D. F., Lever, J.E., and Wright, E. M. (1994) J. Biol. Chem. 269, 22488-22491), we have reevaluated the properties of SGLT2 in greater detail. We furthermore report new data on the regulation of SGLT1 and SGLT2 during kidney development. To analyze and compare SGLT1 and SGLT2 in adult and embryonic kidney, we have cloned and characterized SGLT2 from rat kidney and determined its tissue distribution based on Northern analysis and in situ hybridization. When expressed in Xenopus oocytes, rat SGLT2 stimulated transport of alpha-methyl-D-glucopyranoside (2 mM) in oocytes up to 4.5-fold over controls with an apparent Km of 3.0 mM. The transport properties (i.e. a Na+ to glucose coupling of 1:1 and lack of galactose transport) generally matched those of the kidney cortical low affinity system. We show that expression of rat SGLT2 mRNA is kidney specific and that it is strongly and exclusively expressed in proximal tubule S1 segments. Hybrid-depletion studies were performed to conclusively determine whether SGLT2 corresponds to the kidney cortical low affinity system. Injection of rat kidney superficial cortex mRNA into oocytes stimulated the uptake of alpha-methyl-D-glucopyranoside (2 mM) 2-3-fold. We show that hybrid depletion of this kidney RNA using an SGLT2 antisense oligonucleotide completely suppresses the uptake. These data strongly indicate that SGLT2 is the major kidney cortical low affinity glucose transporter. We therefore propose that SAAT pSGLT2 be renamed SGLT3. Experiments addressing the expression of SGLT1 and SGLT2 mRNAs in embryonic rat kidneys reveal that the two Na+/glucose cotransporters are developmentally regulated and that there may be a different splice variant for SGLT2 in embryonic kidney compared to the adult. PMID- 7493973 TI - Expression of human thyrotropin in cell lines with different glycosylation patterns combined with mutagenesis of specific glycosylation sites. Characterization of a novel role for the oligosaccharides in the in vitro and in vivo bioactivity. AB - We used a novel approach to study the role of the Asn-linked oligosaccharides for human thyrotropin (hTSH) activity. Mutagenesis of Asn (N) within individual glycosylation recognition sequences to Gln (Q) was combined with expression of wild type and mutant hTSH in cell lines with different glycosylation patterns. The in vitro activity of hTSH lacking the Asn alpha 52 oligosaccharide (alpha Q52/TSH beta) expressed in CHO-K1 cells (sialylated oligosaccharides) was increased 6-fold compared with wild type, whereas the activities of alpha Q78/TSH beta and alpha/TSH beta Q23 were increased 2-3-fold. Deletion of the Asn alpha 52 oligosaccharide also increased the thyrotropic activity of human chorionic gonadotropin, in contrast to previous findings at its native receptor. The in vitro activity of wild type hTSH expressed in CHO-LEC2 cells (sialic acid deficient oligosaccharides), CHO-LEC1 cells (Man5GlcNAc2 intermediates), and 293 cells (sulfated oligosaccharides) was 5-8-fold higher than of wild type from CHO K1 cells. In contrast to CHO-K1 cells, there was no difference in the activity between wild type and selectively deglycosylated mutants expressed in these cell lines. Thus, in hTSH, the oligosaccharide at Asn alpha 52 and, specifically, its terminal sialic acid residues attenuate in vitro activity, in contrast to the previously reported stimulatory role of this chain for human chorionic gonadotropin and human follitropin activity. The increased thyrotropic activity of alpha Q52/CG beta suggests that receptor-related mechanisms may be responsible for these differences among the glycoprotein hormones. Despite their increased in vitro activity, alpha Q52/TSH beta, and alpha Q78/TSH beta from CHO-K1 cells had a faster serum disappearance rate and decreased effect on T4 production in mice. These findings highlight the importance of individual oligosaccharides in maintaining circulatory half-life and hence in vivo activity of hTSH. PMID- 7493974 TI - A p53-independent pathway for activation of WAF1/CIP1 expression following oxidative stress. AB - Incubating human cells in diethylmaleate (DEM) depletes the intracellular pool of reduced glutathione (GSH) and increases the concentration of oxidative free radicals. We found that DEM-induced oxidative stress reduced the ability of p53 to bind its consensus recognition sequence and to activate transcription of a p53 specific reporter gene. Nevertheless, DEM treatment induced expression of WAF1/CIP1 but not GADD45 mRNA. The fact that N-acetylcysteine, a precursor of GSH that blocks oxidative stress, prevented WAF1/CIP1 induction by DEM suggests that WAF1/CIP1 induction probably was a consequence of the ability of DEM to reduce intracellular GSH levels. DEM induced WAF1/CIP1 expression in Saos-2 and T98G cells, both of which lack functional p53 protein. DEM treatment did not produce an increase in membrane-associated protein kinase C, but ERK2, a mitogen activated protein kinase, was phosphorylated in a manner consistent with ERK2 activation. DEM treatment also produced a dose-dependent delay in cell cycle progression, which at low concentrations (0.25 mM) consisted of a G2/M arrest and at higher concentrations (1 mM) also involved G1 and S phase delays. Our results indicate that oxidative stress induces WAF1/CIP1 expression and arrests cell cycle progression through a mechanism that is independent of p53. This mechanism may provide for cell cycle checkpoint control under conditions that inactivate p53. PMID- 7493975 TI - An abnormal fibrinogen Fukuoka II (Gly-B beta 15-->Cys) characterized by defective fibrin lateral association and mixed disulfide formation. AB - A dysfibrinogenemia was attributable to a single amino acid substitution from glycine to cysteine at residue 15 of the B beta chain in a fibrinogen molecule designated as fibrinogen Fukuoka II. The fibrinogen Fukuoka II showed prolonged thrombin and reptilase times and impaired fibrinopeptide B release by thrombin, resulting in abolition of fibrin monomer repolymerization under physiological conditions. Repolymerization of the des-(B beta 1-42)-fibrin monomers, however, was not distinguished from the normal pattern of des-(B beta 1-42)-fibrin monomers, suggesting that no other abnormality existed in fibrinogen Fukuoka II. Although an additional cysteine was substituted at residue 15 of the B beta chain, fibrinogen Fukuoka II had no free sulfhydryl group within the molecule. Instead, fibrinogen Fukuoka II formed a disulfide bond with cysteine, albumin, another mutated B beta chain within the same molecule, or intermolecular dimeric fibrinogen Fukuoka II. The mutation in fibrinogen Fukuoka II was the same as that in fibrinogen Ise published previously (Yoshida, N., Wada, H., Morita, K., Hirata, H., Matsuda, M., Yamazumi, K., Asakura, S., and Shirakawa, S. (1991) Blood 77, 1958-1963). Fibrinogen Ise, however, has been described as having prolonged thrombin time but normal reptilase time. Reasons for the discrepancy were not clear. Analysis of the B beta 1-42 fragment showed that fibrinogen was heterogeneous at position 31 of the B beta chain with respect to proline or hydroxyproline. PMID- 7493976 TI - Effect of transcription inhibitors on the iron-dependent degradation of transferrin receptor mRNA. AB - Transferrin receptor (TfR) mRNA expression is tightly linked to intracellular iron levels. Upon iron deprivation, the iron regulatory protein (IRP) stabilizes TfR mRNA by binding to stem-loop structures in its 3'-untranslated region, whereas increased iron levels result in inactivation of the mRNA-binding protein and rapid degradation of TfR mRNA. Although IRP and the regulation of its RNA binding activity have been studied intensively, little is known about the mechanism of TfR mRNA degradation. In order to get more information about factors involved in this process we investigated the in vivo IRP-RNA interaction and the effect of transcription inhibitors on the iron-dependent decay of TfR mRNA. Here we demonstrate that part of the active IRP co-localizes with TfR mRNA to the rough endoplasmic reticulum. High intracellular iron levels led to a drastic reduction of this active RNA-bound IRP in vivo, indicating that IRP dissociates prior to TfR mRNA decay. Furthermore, the transcription inhibitor actinomycin D and translation inhibitor cycloheximide suppressed TfR mRNA degradation but did not interfere with the IRP dissociation step. Other inhibitors of RNA polymerase II had no effect on iron-dependent degradation of TfR mRNA. However, high concentrations of alpha-amanitin known to block transcription by RNA polymerase III interfered with mRNA decay suggesting the involvement of polymerase III transcripts in the degradation pathway. PMID- 7493977 TI - The essential arginine residue at position 210 in the alpha subunit of the Escherichia coli ATP synthase can be transferred to position 252 with partial retention of activity. AB - The substitution of arginine at position 210 in the alpha subunit of Escherichia coli F0F1-ATPase by either lysine or alanine causes dominance in complementation tests with a chromosomal c subunit mutation. Reversal of dominance was achieved for the alpha R210K mutation but not for the alpha R210A mutation by the presence of an aspartic acid residue at position 50 or at position 252 in the alpha subunit. It was concluded that position 210 in putative helix 4 of a previously proposed model of the alpha subunit is close to position 252 in putative helix 5 and to position 50 in putative helix 1. The juxtaposition of residues 252 and 210 was also indicated by the observation that the double mutant alpha R210Q/Q252R was partially functional. A revertant of the partially functional double mutant, isolated on succinate medium, was found to contain a third mutation resulting in Pro-204 in the alpha subunit being replaced by threonine. That the revertant phenotype was due to the alpha P204T change was confirmed by site-directed mutagenesis. ATP synthesis in the revertant strain was at near normal levels as judged by growth yield experiments, but the revertant strain was unable to pump protons in response to ATP hydrolysis. PMID- 7493978 TI - Colocalization of the homophilic binding site and the neuritogenic activity of the cell adhesion molecule L1 to its second Ig-like domain. AB - The cell adhesion molecule L1 has been implicated in mediating cell-cell adhesion and in promoting neurite outgrowth. The extracellular region of L1 contains six immunoglobulin (Ig)-like domains in the amino-terminal region, followed by five fibronectin type III-like repeats. L1 is capable of undergoing homophilic binding as well as heterophilic interactions. To map the homophilic binding domain in L1, three glutathione S-transferase (GST) fusion proteins (GST-Ig1-2-3, GST-Ig4-5-6, and GST-Fn) were prepared and coupled to Covaspheres and their homophilic binding activity was determined using the Covasphere-to-substratum binding assay. Only GST-Ig1-2-3 was capable of homophilic binding. Next, His-tagged recombinant Ig domain proteins (His-Ig1-2, His-Ig1, and His-Ig2) were expressed and subjected to similar assays. Only His-Ig1-2 and His-Ig2 were capable of homophilic interactions. Binding of His-Ig2-conjugated Covaspheres to substrate-coated His Ig2 was inhibited by anti-Ig1-2-3 Fab and soluble His-Ig2. These results indicate that the L1 homophilic binding site resides within Ig2. To examine effects of these L1 recombinant proteins on neurite outgrowth, neural retinal cells were cultured on different substrate-coated fusion proteins. Both GST-Ig1-2-3 and His Ig2 were potent inducers of neurite extension. These results thus indicate that the L1 Ig-like domain 2 alone is sufficient to mediate L1-L1 interaction and promote neurite outgrowth from retinal cells. PMID- 7493979 TI - Hairpin orientation of sterol regulatory element-binding protein-2 in cell membranes as determined by protease protection. AB - Sterol regulatory element-binding proteins (SREBP-1 and SREBP-2) are proteins of approximately 1150 amino acids each that are attached to membranes of the endoplasmic reticulum (ER). In sterol-depleted cells, a protease releases an NH2 terminal fragment of approximately 500 amino acids that contains a basic helix loop-helix leucine zipper motif. This fragment enters the nucleus and stimulates transcription of genes encoding the low density lipoprotein receptor and enzymes of cholesterol biosynthesis. Prior evidence indicates that the SREBPs are attached to membranes by virtue of an 80-residue segment located approximately 80 amino acids to the COOH-terminal side of the leucine zipper. This segment contains two long hydrophobic sequences separated by a short hydrophilic sequence of approximately 30 amino acids. We have proposed a hairpin model in which the two hydrophobic sequences span the membrane, separated by the short hydrophilic sequence which projects into the lumen of the ER (the "lumenal loop"). The model predicts that the NH2- and COOH-terminal segments face the cytosol. To test this model, we constructed a cDNA encoding human SREBP-2 with epitope tags at the NH2 terminus and in the lumenal loop. The COOH-terminal region was visualized with a newly developed monoclonal antibody against this region. Sealed membrane vesicles were isolated from cells expressing the epitope-tagged version of SREBP-2. Trypsin treatment of these vesicles destroyed the NH2- and COOH-terminal segments and reduced the lumenal epitope to a size consistent with protection of the lumenal sequence plus the two membrane-spanning segments. The lumenal epitope tag contained two potential sites for N-linked glycosylation. The size of the trypsin protected fragment was reduced by treatment with N-Glycanase and endoglycosidase H, indicating that this segment was located in the lumen of the ER where it was glycosylated. These data provide strong support for the hairpin model. PMID- 7493980 TI - The beta subunit Rif-cluster I is only angstroms away from the active center of Escherichia coli RNA polymerase. AB - Ribonucleotide analogs bound in the initiating site of Escherichia coli RNA polymerase-promoter complex were cross-linked to the beta subunit. Using limited proteolysis and chemical degradation, the cross-link was mapped to a segment of beta between amino acids Val516 and Arg540. This region (Rif-cluster I) is known to harbor many rifampicin-resistant (RifR) mutations. The results demonstrate that Rif-culster I is part of the "5'-face" of the active center and provide structural basis for the long-known effects of RifR mutations on transcription initiation, elongation, and termination. PMID- 7493981 TI - Hsp90 mutants disrupt glucocorticoid receptor ligand binding and destabilize aporeceptor complexes. AB - In order to attain competence to respond to hormone, certain steroid hormone receptors must be assembled into hetero-oligomeric aporeceptor complexes, containing Hsp90 and other proteins. Members of the Hsp90 gene family are highly conserved, strongly expressed, and required for viability in eukaryotic organisms. To elucidate the role of Hsp90 in the activity of steroid hormone receptors in vivo, four Hsp90 mutatns, which cause defects in glucocorticoid receptor (GR) signaling, but support the viability of Saccharomyces cerevisiae, were previously isolated (Bohen, S. P., and Yamamoto, K. R. (1993) Proc. Natl. Acad. Sci. U.S.A. 90, 11424-11428). In this study, I characterize the effects of the Hsp90 mutants on GR ligand response, ligand binding activity, and aporeceptor complex stability. The mutants fall into two classes. Three of the Hsp90 mutants cause defects in GR ligand binding in vivo and form aporeceptor complexes that are unstable in vitro, relative to those containing wild-type Hsp90. The other mutant affects GR signaling, but aporeceptor complexes with this mutant are not defective for ligand binding or stability. These findings indicate that the binding of Hsp90 to GR in the aporeceptor complex is insufficient to induce a high ligand affinity conformation, rather the high ligand affinity to GR requires a specific interaction with Hsp90, which is altered by certain Hsp90 mutants. PMID- 7493982 TI - Molecular cloning, expression, and characterization of podocalyxin-like protein 1 from rabbit as a transmembrane protein of glomerular podocytes and vascular endothelium. AB - Podocytes are responsible in part for maintaining the size and charge filtration characteristics of the glomerular filter. The major sialoprotein of the podocyte foot process glycocalyx is a 140-kDa sialoprotein named podocalyxin. Monoclonal antibodies raised against isolated rabbit glomeruli that recognized a podocalyxin like protein base upon size, Alcian blue staining, wheat germ agglutinin binding, and distribution in renal cortex were used to expression clone cDNAs from a rabbit glomerular library. On Northern blot the cDNAs hybridized to a 5.5 kilobase pair transcript predominantly present in glomerulus. The overlapping cDNAs spanned 5,313 base pairs that contained an open reading frame of 1,653 base pairs and were not homologous with a previously described sequence. The deduced 551-amino acid protein contained a putative 21-residue N-terminal signal peptide and a 26-amino acid transmembrane region. The mature protein has a calculated molecular mass of 55 kDa, an extracellular domain that contains putative sites for N- and O-linked glycosylation, and a potential glycosaminoglycan attachment sites. The intracellular domain contains potential sites for phosphorylation. Processing of the full-length coding region in COS-7 cells resulted in a 140-kDa band, suggesting that the 55-kDa core protein undergoes extensive post translational modification. The relationship between the cloned molecule and the monoclonal antibodies used for cloning was confirmed by making a fusion protein that inhibited binding of the monoclonal antibodies to renal cortical tissue sections and then raising polyclonal antibodies against the PCLP1 fusion protein that also recognized glomerular podocytes and endothelial cells in tissue sections in a similar distribution to the monoclonal antibodies. We conclude that we have cloned and sequenced a novel transmembrane core glycoprotein from rabbit glomerulus, which has many of the characteristics of podocalyxin. We have named this protein podocalyxin-like protein 1. PMID- 7493983 TI - Degradation process of ligand-stimulated platelet-derived growth factor beta receptor involves ubiquitin-proteasome proteolytic pathway. AB - The platelet-derived growth factor beta-receptor undergoes polyubiquitination as a consequence of ligand binding. We have previously reported that ligand-induced ubiquitination of the receptor plays a negative regulatory role in its mitogenic signaling possibly by promoting the efficient degradation of the ligand-activated receptor (Mori, S., Heldin, C.-H., and Claesson-Welsh, L. (1993) J. Biol. Chem. 268, 577-583). In the present study, we have examined effects of different kinds of cell-penetrating proteasome inhibitors, including substrate-related peptidyl aldehydes, Cbz-Ile-Glu(O-t-Bu)-Ala-leucinal (where Bu is butyl and Cbz is benzyloxycarbonyl) (PSI) and Cbz-Leu-Leu-norvalinal (MG115), and a Streptomyces metabolite lactacystin, on degradation of the receptor in intact cells with the aim of evaluating the role of the receptor ubiquitination in the proteasome dependent proteolytic process. These proteasome inhibitors were found to considerably inhibit ligand-stimulated degradation of the wild-type beta receptor; however, their inhibitory effect was not observed when the cells expressing the ubiquitination-deficient mutant beta-receptor were analyzed. These data suggest that the degradation process of the ligand-stimulated beta-receptor involves the ubiquitin-proteasome proteolytic pathway. PMID- 7493984 TI - The isolation and characterization of cDNA encoding the mouse bifunctional ATP sulfurylase-adenosine 5'-phosphosulfate kinase. AB - Biosynthesis of the activated sulfate donor, adenosine 3'-phosphate 5' phosphosulfate, involves the sequential action of two enzyme activities: ATP sulfurylase, which catalyzes the formation of adenosine 5'-phosphosulfate (APS) from ATP and free sulfate, and APS kinase, which subsequently phosphorylates APS to produce adenosine 3'-phosphate 5'-phosphosulfate. Oligonucleotide primers were derived from a human infant brain-expressed sequence tag putatively encoding a portion of APS kinase. Using these primers, reverse transcriptase-polymerase chain reaction was performed on mRNA from neonatal normal mice resulting in amplification of a 127-bp DNA fragment. This fragment was subsequently used to screen a mouse brain lambda gt11 cDNA library, yielding a 2.2-kb clone. Primers were designed from the 5'-end of the 2.2-kb clone, and 5'-rapid amplification of cDNA ends was used to obtain the translation start site. Sequence from the overlapping clones was assembled into a 2475-bp composite sequence, which contains a single open reading frame that translates into a 624-deduced amino acid sequence. Northern blots of total RNA from neonatal mice yielded a single message species at approximately 3.3 kb. Southern blot of genomic DNA digested with several restriction enzymes suggested the gene is present as a single copy. Comparison against sequence data bases suggested the composite sequence was a fused sulfurylase-kinase product, since the deduced amino acid sequence showed extensive homology to known separate sequences of both ATP sulfurylase and APS kinase from several sources. The first 199 amino acids corresponded to APS kinase sequence, followed by 37 distinct amino acids, which did not match any known sequence, followed by 388 amino acids that are highly homologous to known ATP sulfurylase sequences. Finally, recombinant enzyme expressed in COS-1 cells exhibited both ATP sulfurylase and APS kinase activity. PMID- 7493986 TI - Primary structure of a gamma subunit of G protein, gamma 12, and its phosphorylation by protein kinase C. AB - We have determined the primary structure of a novel gamma subunit (gamma 12, previously designated gamma S1) of G protein purified from bovine spleen. The mature gamma 12 protein composed of 68 amino acids had acetylated serine at the N terminus and geranylgeranylated/carboxylmethylated cysteine at the C terminus. This was consistent with the C-terminal prenylation signal in the amino acid sequence, which was predicted from gamma 12 cDNA isolated from a bovine spleen cDNA library. Western blots with the specific antibody against gamma 12 showed that gamma 12 is present in all tissues examined. Among various gamma subunits (gamma 1, gamma 2, gamma 3, gamma 7, and gamma 12), gamma 12 has a unique property to be phosphorylated by protein kinase C. The phosphorylated amino acid residue was Ser1 (or Ser2). The phosphorylated beta gamma 12 associated with Go alpha more tightly than the unphosphorylated form. Exposure of Swiss 3T3 and aortic smooth muscle cells to phorbol 12-myristate 13-acetate and NaF induced phosphorylation of gamma 12. Stimulation of aortic smooth muscle cells with natural vasoactive agents such as angiotensin II and vasopressin also induced phosphorylation of gamma 12. The extent of phosphorylation of beta gamma 12 in vitro was suppressed by a complex formation with Go alpha, which was relieved by the addition of guanosine 5'-O-(3-thiotriphosphate) or aluminum fluoride. These results strongly suggest that gamma 12 is phosphorylated by protein kinase C during activation of receptor(s) and G protein(s) in living cells. PMID- 7493985 TI - Characterization of the promoter region of the human transforming growth factor beta type II receptor gene. AB - Diminished cellular responsiveness to transforming growth factor-beta (TGF-beta) is frequently correlated with decreased transcription of the type II receptor for TGF-beta (TGF-beta RII). We have cloned and characterized the human TGF-beta RII promoter and, using S1 nuclease mapping and 5' rapid amplification of cDNA ends polymerase chain reaction, have identified five alternative transcription start sites within the region -33 to +57. DNA transfection experiments and electrophoretic mobility shift assays have revealed the existence of five distinct regulatory regions including two positive regulatory elements and two negative regulatory elements in addition to the core promoter region. The first positive regulatory element (-219 to -172) interacts with two distinct nuclear protein complexes, at least one of which appears to be a previously unidentified transcription factor. The second positive regulatory element (+1 to +35) also interacts with two separate protein complexes, both of which appear to be novel transcription factors. Deletion of either positive regulatory element markedly decreased expression of the target gene, suggesting that both positive regulatory elements are necessary for basal expression levels of TGF-beta RII. PMID- 7493987 TI - Transactivation by Rtg1p, a basic helix-loop-helix protein that functions in communication between mitochondria and the nucleus in yeast. AB - Rtg1p is a basic helix-loop-helix transcription factor in the yeast Saccharomyces cerevisiae that is required for basal and regulated expression of CIT2, the gene encoding a peroxisomal isoform of citrate synthase. In respiratory incompetent rho degree petite cells, CIT2 transcription is elevated as much as 30-fold compared with respiratory competent rho + cells. Here we provide evidence that Rtg1p interacts directly with a CIT2 upstream activation site (UASr) and that the rho degree/rho + regulation is not due to a change in the levels of Rtg1p. A fusion protein consisting of the DNA binding domain of Gal4p fused to the NH2 terminus of the full-length wild-type Rtg1p was able to transactivate an integrated LacZ reporter under control of the Gal4p-responsive GAL1 UASG in a rho degree/rho(+)-dependent manner. Other Gal4p fusions to deletions or mutations of Rtg1p indicate that the helix-loop-helix domain is essential for transactivation. Regulated expression of CIT2 also requires the RTG2 gene product. The Gal4-Rtg1p fusion was unable to transactivate the LacZ reporter gene in a strain deleted for RTG2, suggesting that the RTG2 product does not act independently of Rtg1p in the rho degree/rho + transcriptional response. PMID- 7493989 TI - Active site and oligosaccharide recognition residues of peptide-N4-(N-acetyl-beta D-glucosaminyl)asparagine amidase F. AB - Crystallographic analysis and site-directed mutagenesis have been used to identify the catalytic and oligosaccharide recognition residues of peptide-N4-(N acetyl-beta-D-glucosaminyl)asparagine amidase F (PNGase F), an amidohydrolase that removes intact asparagine-linked oligosaccharide chains from glycoproteins and glycopeptides. Mutagenesis has shown that three acidic residues, Asp-60, Glu 206, and Glu-118, that are located in a cleft at the interface between the two domains of the protein are essential for activity. The D60N mutant has no detectable activity, while E206Q and E118Q have less than 0.01 and 0.1% of the wild-type activity, respectively. Crystallographic analysis, at 2.0-A resolution, of the complex of the wild-type enzyme with the product, N,N'-diacetylchitobiose, shows that Asp-60 is in direct contact with the substrate at the cleavage site, while Glu-206 makes contact through a bridging water molecule. This indicates that Asp-60 is the primary catalytic residue, while Glu-206 probably is important for stabilization of reaction intermediates. Glu-118 forms a hydrogen bond with O6 of the second N-acetylglucosamine residue of the substrate and the low activity of the E118Q mutant results from its reduced ability to bind the oligosaccharide. This analysis also suggests that the mechanism of action of PNGase F differs from those of L-asparaginase and glycosylasparaginase, which involve a threonine residue as the nucleophile. PMID- 7493988 TI - Targeted disruption of the beta 3-adrenergic receptor gene. AB - beta 3-Adrenergic receptors (beta 3-ARs) are expressed predominantly in white and brown adipose tissue, and beta 3-selective agonists are potential anti-obesity drugs. However, the role of beta 3-ARs in normal physiology is unknown. To address this issue, homologous recombination was used to generate mice that lack beta 3-ARs. This was accomplished by direct injection of a DNA-targeting construct into mouse zygotes. Twenty-three transgenic mice were generated, of which two had targeted disruption of the beta 3-AR gene. Mice that were homozygous for the disrupted allele had undetectable levels of intact beta 3-AR mRNA, as assessed by RNase protection assay and Northern blotting, and lacked functional beta 3-ARs, as demonstrated by complete loss of beta 3-agonist (CL 316,243)-induced stimulation of adenylate cyclase activity and lipolysis. beta 3 AR-deficient mice had modestly increased fat stores (females more than males), indicating that beta 3-ARs play a role in regulating energy balance. Importantly, beta 1 but not beta 2-AR mRNA levels up-regulated in white and brown adipose tissue of beta 3-AR-deficient mice (brown more than white), strongly implying that beta 3-ARs mediate physiologically relevant signaling under normal conditions and that "cross-talk" exists between beta 3-ARs and beta 1-AR gene expression. Finally, acute treatment of normal mice with CL 316,243 increased serum levels of free fatty acids (FFAs) (3.2-fold) and insulin (140-fold), increased energy expenditure (2-fold), and reduced food intake (by 45%). These effects were completely absent in beta 3-AR-deficient mice, proving that the actions of CL are mediated exclusively by beta 3-ARs. beta 3-AR-deficient mice should be useful as a means to a better understanding of the physiology and pharmacology of beta 3-ARs. PMID- 7493990 TI - Structural and functional consequences of mutations in 6-pyruvoyltetrahydropterin synthase causing hyperphenylalaninemia in humans. Phosphorylation is a requirement for in vivo activity. AB - Four naturally occurring mutants with single amino acid alterations in human 6 pyruvoyltetrahydropterin synthase (PTPS) were overexpressed and characterized in vitro. The corresponding DNA mutations were found in patients with hyperphenylalaninemia and monoamine neurotransmitter insufficiency due to lack of the tetrahydrobiopterin biosynthetic enzyme PTPS. To predict the structure of the mutant enzymes, computer modeling was performed based on the solved three dimensional structure of the homohexameric rat enzyme. One mutant (delta V57) is incorrectly folded and thus unstable in vitro and in vivo, while a second mutant (P87L) has substantial activity but enhanced sensitivity to local unfolding. Two other mutants, R16C and R25Q, form stable homomultimers and exhibit significant activity in vitro but no activity in COS-1 cells. In vivo 32P labeling showed that wild-type PTPS, P87L, and R25Q are phosphorylated, while R16C is not modified. This strongly suggests that the serine 19 within the consensus sequence for various kinases, RXXS, is the site of modification. Our results demonstrate that PTPS undergoes protein phosphorylation and requires additional, not yet identified post-translational modification(s) for its in vivo function. PMID- 7493993 TI - Crystallographic studies on the binding modes of P2-P3 butanediamide renin inhibitors. AB - The binding modes of three peptidomimetic P2-P3 butanediamide renin inhibitors have been determined by x-ray crystallography. The inhibitors are bound with their backbones in an extended conformation, and their side chains occupying the S5 to S1' pockets. A (2-amino-4-thiazolyl)methyl side chain at the P2 position shows stronger hydrogen-bonding and van der Waals interactions with renin than the His side chain, which is present in the natural substrate. The ACHPA-gamma lactam transition state analog has similar interactions with renin as the dihydroxyethylene transition state analog. PMID- 7493991 TI - Organization and analysis of the complete rat calmodulin-dependent protein kinase IV gene. AB - A 42-kilobase pair region of rat DNA containing the Ca2+/calmodulin-dependent protein kinase IV (CaM kinase IV) gene has been cloned and characterized. The gene consists of 12 exons and 11 introns and is predicted to encode both beta and alpha forms of CaM kinase IV as well as the testis-specific calmodulin-binding protein calspermin. The promoter utilized to generate the alpha-kinase isoform is located in intron 1, whereas the promoter utilized to produce the calspermin transcript is contained in intron 10. The calspermin promoter region which extends from -200 to +321 relative to the calspermin transcription initiation site that contains two cyclic AMP response elements (CRE) at -70 and -50 and has been shown previously to be inactive in NIH3T3 cells (Sun, Z., Sassone-Corsi, P., and Means, A. R. (1995) Mol. Cell. Biol. 15, 561-571) was ligated to the lacZ reporter gene and used to generate transgenic mice. The promoter was expressed exclusively in postmeiotic testis where beta-galactosidase was found predominantly in elongating spermatids. The cell and developmental specificity of transgene expression was very similar to the pattern shown by the endogenous gene. Although the transgene promoter was silent in somatic tissues, beta galactosidase expression could be restored in primary cultures of skin fibroblasts by introduction of vectors encoding CREM tau and CaM kinase IV. PMID- 7493992 TI - Remodeling of mouse milk glycoconjugates by transgenic expression of a human glycosyltransferase. AB - The mammary gland is a unique biosynthetic tissue that produces a variety of species-specific glycoconjugates, but the factors regulating the production of specific glycoconjugates are not well understood. To explore the underlying regulation, a fusion gene containing a cDNA encoding the human alpha 1,2 fucosyltransferase (alpha 1,2FT), which generates the H-blood group antigen, flanked by the murine whey acidic protein promoter and a polyadenylation signal, was introduced into mice. Milk samples from transgenic animals contained soluble forms of the alpha 1,2FT, as revealed by Western blots of milk samples using an anti-alpha 1,2FT antiserum and by the demonstration of alpha 1,2FT enzyme activity. Milk from transgenic animals also contained large quantities of 2' fucosyllactose (Fuc alpha 1-2Gal beta 1-4Glc) and modified glycoproteins containing the H-antigen, whereas milk from control animals lacked these glycoconjugates. Expression levels of 2'-fucosyllactose were high in most animals and represented 1/3 to nearly 1/2 of the total milk oligosaccharides. These results demonstrate that heterologous transgenic expression of a glycosyltransferase can result in the expression of both the transgene and its secondary gene products and that the structures of milk oligosaccharides can be remodeled depending on expression of the appropriate enzyme. Furthermore, these results suggest that the lactating mammary gland may be a unique biosynthetic reactor for the production of biologically active oligosaccharides and glycoconjugates. PMID- 7493994 TI - Phosphorylation of the platelet p47 phosphoprotein is mediated by the lipid products of phosphoinositide 3-kinase. AB - Platelet stimulation by thrombin or the thrombin receptor activating peptide (TRAP) results in the activation of phosphoinositide 3-kinase and the production of the novel polyphosphoinositides phosphatidylinositol 3,4-bisphosphate (PtdIns 3,4-P2) and phosphatidylinositol 3,4,5-trisphosphate (PtdIns-3,4,5-P3). We have shown previously that these lipids activate calcium-independent protein kinase C (PKC) isoforms in vitro (Toker, A., Meyer, M., Reddy, K. K., Falck, J. R., Aneja, R., Aneja, S., Parra, A., Burns, D. J., Ballas, L. M. and Cantley, L. C. (1994) J. Biol. Chem. 269, 32358-32367). Activation of platelet PKC in response to TRAP is detected by the phosphorylation of the major PKC substrate in platelets, the p47 phosphoprotein, also known as pleckstrin. Here we provide evidence for two phases of pleckstrin phosphorylation in response to TRAP. A rapid phase of pleckstrin phosphorylation (< 1 min) precedes the peak of PtdIns-3,4-P2 production and is unaffected by concentrations of wortmannin (10-100 nM) that block production of this lipid. However prolonged phosphorylation of pleckstrin (> 2 min) is inhibited by wortmannin concentrations that block PtdIns-3,4-P2 production. Phorbol ester-mediated pleckstrin phosphorylation was not affected by wortmannin and wortmannin had no effect on purified platelet PKC activity. Phosphorylation of pleckstrin could be induced using permeabilized platelets supplied with exogenous gamma-32P[ATP] and synthetic dipalmitoyl PtdIns-3,4,5-P3 and dipalmitoyl PtdIns-3,4-P2 micelles, but not with dipalmitoyl phosphatidylinositol 3-phosphate or phosphatidylinositol 4,5-bisphosphate. These results suggest two modes of stimulating pleckstrin phosphorylation: a rapid activation of PKC (via diacylglycerol and calcium) followed by a slower activation of calcium-independent PKCs via PtdIns-3,4-P2. PMID- 7493995 TI - Regulation and immunolocalization of acyl-coenzyme A: cholesterol acyltransferase in mammalian cells as studied with specific antibodies. AB - Acyl-coenzyme A:cholesterol acyltransferase (ACAT) catalyzes the formation of intracellular cholesterol esters in various tissues. We recently reported the cloning and expression of human macrophage ACAT cDNA. In the current study, we report the production of specific polyclonal antibodies against ACAT by immunizing rabbits with the recombinant fusion protein composed of glutathione S transferase and the first 131 amino acids of ACAT protein. Immunoblot analysis showed that the antibodies cross-reacted with a 50-kDa protein band from a variety of human cell lines. These antibodies immunodepleted more than 90% of detergent-solubilized ACAT activities from six different human cell types, demonstrating that the 50-kDa protein is the major ACAT catalytic component in these cells. In multiple human tissues examined, the antibodies recognized protein bands with various molecular weights. These antibodies also cross-reacted with the ACAT protein in Chinese hamster ovary cells. Immunoblot analysis showed that the ACAT protein contents in human fibroblast cells, HepG2 cells, or Chinese hamster ovary cells were not affected by sterol in the medium, demonstrating that the main mechanism for sterol-dependent regulation of ACAT activity in these cells is not change in ACAT protein content. As revealed by indirect immunofluorescent microscopy, the ACAT protein in tissue culture cells was located in the endoplasmic reticulum. This finding, along with earlier studies, suggests that cholesterol concentration in the endoplasmic reticulum may be the major determinant for regulating ACAT activity in the intact cells. PMID- 7493996 TI - The cytokine responsive vascular smooth muscle cell enhancer of inducible nitric oxide synthase. Activation by nuclear factor-kappa B. AB - The production of inducible nitric oxide synthase (iNOS) within vascular smooth muscle (VSM) cells following exposure to proinflammatory cytokines is a major cause of the vasorelaxation and hypotension of septic shock. We have defined the cytokine-responsive element of the murine iNOS promoter, transfected into a VSM cell line, and the role of the NF-kappa B/Rel family of proteins in iNOS gene activation in these cells. The combination of interleukin-1, interferon-gamma, and tumor necrosis factor-alpha stimulates promoter activity by a factor of 8.1 fold; single cytokines show little activity, while pairs of cytokines produce an intermediate effect. Using a series of promoter deletion mutants, we have defined the cytokine-responsive element from position -890 to -1002; this region contains an NF-kappa B-binding site as well as a number of interferon response elements. Nuclear proteins from cytokine-stimulated VSM cells which bind to an oligonucleotide containing this kappa B site are composed of p65 together with an unidentified protein of 50 kDa, which is not a known Rel family member. A promoter mutant with a 2-base pair change within this kappa B site, which abolishes NF-kappa B binding, has an activity of only approximately 34% (S.E. +/- 1.5) of the wild-type promoter. In addition, protein binding to this site is abolished by a specific inhibitor of NF-kappa B activation, which also abrogates iNOS activity. Residual inducibility in such mutant promoters is attributable to the presence of an independently functioning downstream kappa B site (-85 to 75). The mechanism by which NF-kappa B activates the iNOS promoter in VSM cells in response to cytokines appears to be markedly different to that operative in macrophages in response to lipopolysaccharide. PMID- 7493997 TI - Translocation of loops regulates transport activity of mitochondrial ADP/ATP carrier deduced from formation of a specific intermolecular disulfide bridge catalyzed by copper-o-phenanthroline. AB - The cross-linking reagent copper-o-phenanthroline complex (Cu(OP)2) specifically caused a decrease in the amount of the 30-kDa ADP/ATP carrier in bovine submitochondrial particles associated predominantly with formation of a 60-kDa protein consisting of a cross-linked dimer of the carrier. However, Cu(OP)2 had no effect on mitochondria. The transport of ADP via the carrier through submitochondrial particle membranes was found to be inhibited in parallel with the progress of intermolecular cross-linking. Analysis of the cross-linked site showed that a disulfide bridge was formed only between two Cys56 residues in a pair of the first loops facing the matrix space. The transport inhibitor bongkrekic acid, which locks the m-state conformation of the carrier, had no effect on disulfide bridge formation catalyzed by Cu(OP)2, but carboxyatractyloside, which locks the c-state conformation by acting from the cytosolic side, completely inhibited the cross-linking. These results show that the ADP/ATP carrier functions as a dimer form, and a pair of the first loops protrudes into the matrix space in the m-state, but possibly intrudes into the membrane in the c-state. Thus, it is suggested that a pair of the first loops acts as a gate and that its opening and closing are regulated by their translocation. PMID- 7493998 TI - DnaX complex of Escherichia coli DNA polymerase III holoenzyme. Central role of tau in initiation complex assembly and in determining the functional asymmetry of holoenzyme. AB - The alternative forms of the DnaX protein found in Escherichia coli DNA polymerase III holoenzyme, tau and gamma, were purified from extracts of strains carrying overexpressing plasmids mutated in their frameshifting sequences such that they produced only one subunit or the other. The purified subunits were used to reconstitute the tau and gamma complexes which were characterized by functional assays. The gamma complex-reconstituted holoenzyme required a stoichiometric excess of DNA polymerase III core, beyond physiological levels, for activity. The tau subunit stimulated the gamma complex 2-fold, but could not be used to reconstitute a holoenzyme with gamma complex and stoichiometric quantities of core. In the presence of adenosine 5'-O-(3'-thiotriphospate) (ATP gamma S), the DNA polymerase III holoenzyme behaves as an asymmetric dimer; it can form only initiation complexes with primed DNA in one-half of the enzyme (Johanson, K. O., and McHenry, C. S. (1984) J. Biol. Chem. 259, 4589-4595). An asymmetric distribution of two products of the dnaX gene, gamma and tau, has been postulated to underlie the asymmetry of holoenzyme. To provide a direct test for this hypothesis, we reconstituted holoenzyme containing only the gamma or tau DnaX proteins. We observed that, although gamma could function in the presence of ATP and high concentrations of DNA polymerase III core, it was nearly inert in the presence of ATP gamma S. In contrast, tau-containing holoenzyme behaved exactly like native holoenzyme in the presence of ATP gamma S. These results implicate tau as a key component required to reconstitute holoenzyme with native behavior and show that tau plays a key role in initiation complex formation. These results also show that gamma is not a necessary component, since all of the known properties of native holoenzyme can be reproduced with a 9-subunit tau holoenzyme. PMID- 7493999 TI - DnaX complex of Escherichia coli DNA polymerase III holoenzyme. Physical characterization of the DnaX subunits and complexes. AB - A physical characterization of the tau and gamma subunits of the Escherichia coli DNA polymerase III holoenzyme and their complexes with the delta, delta', chi, and psi subunits is presented. The native molecular mass of the tau and gamma subunits was determined to be 255,000 and 189,000 Da, respectively, by sedimentation equilibrium analytical ultracentrifugation. Both values indicate a tetrameric quaternary structure. The tau and gamma complexes were reconstituted and purified using two different methods. Both complexes assembled readily and were reconstituted at subunit concentrations approaching physiological levels. The stoichiometries of the tau and gamma complexes, as determined by quantitative densitometry of SDS-polyacrylamide gels, were found to be tau 4 delta 1 delta' 1 chi 1 psi 1 and gamma 4 delta 1 delta' 1 chi 1 psi 1. BIAcore analysis demonstrated that the formation of large multiprotein complexes of holoenzyme subunits depends on the presence of the tau subunit; gamma could not substitute. We present a model for a gamma-less form of DNA polymerase III holoenzyme that has asymmetrical structural features that may be responsible for the functional asymmetry observed in holoenzyme. The stoichiometry of the reconstituted DNA polymerase III* component of holoenzyme in this model is (alpha epsilon theta)2DnaX4 delta 1 delta' 1 chi 1 psi 1. PMID- 7494000 TI - DnaX complex of Escherichia coli DNA polymerase III holoenzyme. The chi psi complex functions by increasing the affinity of tau and gamma for delta.delta' to a physiologically relevant range. AB - An artificial operon that contains tandem holC-holD genes was used to overproduce a complex of the chi and psi subunits of the DNA polymerase III holoenzyme. Normally insoluble by itself, psi forms a tight soluble complex with chi. A purification procedure that yields pure, active chi psi complex in 100-mg quantities suitable for biophysical studies is reported. Sedimentation equilibrium studies demonstrate that chi psi is a 1:1 heterodimer. The presence of chi psi dramatically lowers the level of delta.delta' required to reconstitute holoenzyme to levels expected in vivo. That chi psi accomplishes this by binding to gamma or tau and increasing their affinity for delta.delta' was demonstrated by surface plasmon resonance using a Pharmacia BIA-core instrument. In the absence of delta.delta', chi psi binds to either the gamma or tau DnaX protein with Kd = 2 nM. PMID- 7494001 TI - Heparin-like molecules on the cell surface potentiate binding of diphtheria toxin to the diphtheria toxin receptor/membrane-anchored heparin-binding epidermal growth factor-like growth factor. AB - Diphtheria toxin receptor (DTR), which is identical to the membrane-anchored form of heparin-binding EGF-like growth factor (proHB-EGF), has a high affinity for heparin. We studied the effect of heparin-like molecules on the binding of diphtheria toxin (DT) to DTR/proHB-EGF. Mutant Chinese hamster ovary (CHO) cells deficient in heparan sulfate (HS) proteoglycans were about 15 times less sensitive to DT than wild type CHO-K1 cells. When free heparan sulfate or heparin was added to the culture medium, DT sensitivity of the mutant cells was fully restored. Studies of binding of 125I-labeled DT to HS-deficient CHO cells transfected with human DTR/proHB-EGF cDNA indicated that the increased sensitivity to DT after addition of heparin is due to increased binding of DT to cells. Vero cells display a relatively large amount of heparan sulfate residues compared to CHO-K1 cells or L cells. Enhancement of DT binding by the addition of heparin was also observed with CHO-K1 cells and L cells that had been transfected with human DTR/proHB-EGF cDNA, but the degree of enhancement was less than that observed with the HS-deficient CHO cells. Addition of heparin did not affect DT binding or DT sensitivity of Vero cells. Heparin-dependent binding was observed when intact Vero cells were treated with heparitinase or when the cell membrane was solubilized with a neutral detergent. Scatchard plot analysis for the binding of DT to a recombinant HB-EGF in vitro and to L cells expressing human DTR/proHB EGF revealed that heparin increases the affinity of DTR/proHB-EGF for DT but does not change the number of binding sites. Although DRAP27/CD9 is known to enhance DT binding to DTR/proHB-EGF, the results indicate that heparin and DRAP27/CD9 increase DT binding by independent mechanisms. Thus, heparin-like molecules, probably in the form of heparan sulfate proteoglycan on the cell surface, are a third factor required for maximal DT binding activity of cells. PMID- 7494002 TI - Characterization of heparan sulfate oligosaccharides that bind to hepatocyte growth factor. AB - Proteoglycans from rat liver had the ability to bind hepatocyte growth factor (HGF). Digestion of the proteoglycans with heparitinase resulted in the complete loss of the activity, while the digestion with chondroitinase ABC had no effect. Heparan sulfate (HS)-conjugated gel also bound HGF, and the binding was competitively inhibited by heparin and bovine liver HS, but not by Engelbreth Holm-Swarm sarcoma HS, pig aorta HS, or other glycosaminoglycans, suggesting the specific structural domain in HS for the binding of HGF. Among limited digests with heparitinase I of bovine liver HS, octasaccharide is the minimal size to bind HGF. Comparison of the disaccharide unit compositions revealed a marked difference in IdoA(2SO4)-GlcNSO3(6SO4) unit between the bound and unbound octasaccharides. The contents of this disaccharide unit were calculated to be 2 mol/mol for the bound octasaccharide but 1 mol/mol for the unbound one. Considering both the substrate specificity and properties of heparitinase I, the above results suggest that the bound octasaccharide should contain two units of IdoA(2SO4)-GlcNSO3(6SO4) contiguously or alternately in the vicinity of the reducing end. The bound decasaccharide was more than 20 times as active as the unbound one with regard to the ability to release HGF bound to rat liver HS proteoglycan. The ability was comparable to the one-fourth of that of heparin. PMID- 7494003 TI - The plasmacytoma growth inhibitor restrictin-P is an antagonist of interleukin 6 and interleukin 11. Identification as a stroma-derived activin A. AB - A stromal protein, designated restrictin-P, that specifically kills plasma-like cells was purified to homogeneity and shown to be identical with activin A. The specificity to plasma-like cells stemmed from the ability of restrictin-P/activin A to competitively antagonize the proliferation-inducing effects of interleukin (IL) 6 and IL-11. Restrictin-P further interfered with the IL-6-induced secretion of acute phase proteins by HepG2 human hepatoma cells and with the IL-6-mediated differentiation of M1 myeloblasts. A competition binding assay indicated that restrictin-P did not interfere with the binding of IL-6 to its receptor on plasma like cells, suggesting that it may act by intervening in the signal transduction pathway of the growth factor. Indeed, concomitant addition of restrictin-P and IL 6 to cytokine-deprived B9 hybridoma cells was followed by sustained overexpression of junB gene until cell death occurred, while IL-6 alone caused a transient increase only. This altered response to IL-6 stimulation was accompanied by a moderate increase in STAT protein activation. Thus, in this study, we identified the plasmacytoma growth inhibitor, restrictin-P, as being activin A of stromal origin. It is shown that activin A is an antagonist of IL-6 induced functions and that it modifies the IL-6 signaling pattern. PMID- 7494004 TI - Interaction of streptokinase and plasminogen. Studied with truncated streptokinase peptides. AB - The interaction of streptokinase (SK) with human plasminogen (HPlg) was investigated using truncated SK peptides prepared by gene cloning techniques. SK(16-414) and SK(16-378) could activate HPlg as efficiently as the authentic SK. SK(60-414), which had been preincubated with SK(1-59), could also activate HPlg. SK(91-414), SK(127-414), and SK(158-414), at a concentration of one-tenth of HPlg, all failed to activate HPlg. However, the truncated SK peptides in complexes with equimolar HPlg could form amidolytically active virgin enzymes that slowly converted to human plasmin (HPlm) after a lag period of 15 min. SK(16 316) could not activate HPlg. No virgin enzyme was detected when SK(16-316) was incubated with equimolar HPlg, but the HPlg in the complex was modified to HPlm after reaction for 20 min. SK(220-414) and SK(16-251) had no ability to transform HPlg to virgin enzyme or to HPlm in equimolar complex with HPlg, although they could bind to HPlg. The functions of five regions in the SK molecule (a, Ile1 Lys59; b, Ser60-Asn90; c, Val158-Arg219; d, Tyr252-Ala316; e, Ser317-Ala378) in interaction with HPlg are deduced. Region a is important in stabilizing the conformation of the SK molecule, and region b is essential for HPlg activation. Region c is required for induction of the conformational changes of HPlg to virgin enzyme. Regions c and d are required for the conversion of HPlg to HPlm in the HPlg.SK equimolar complex. Coordination of regions c, d, and e of SK is essential for a virgin enzyme formation, and coordination of regions b, c, d and e is required for an effective SK-type HPlg activator. PMID- 7494005 TI - Agonist-dependent phosphorylation and desensitization of the rat A3 adenosine receptor. Evidence for a G-protein-coupled receptor kinase-mediated mechanism. AB - A3 adenosine receptor (A3AR) activation contributes to both the cardioprotective and antihypertensive effects of adenosine. To date, no studies have examined the mechanisms by which this receptor undergoes rapid homologous desensitization. Therefore, a functional hemagglutinin epitope-tagged A3AR has been stably expressed in Chinese hamster ovary cells, and its regulation by the AR agonist 5' N-ethylcarboxamidoadenosine (NECA) has been studied. Cellular exposure to NECA induces rapid (t1/2 = approximately 1 min) A3AR phosphorylation on serine and threonine residues. This is associated with a functional desensitization and a 30 40% reduction in the number of high affinity agonist binding sites as determined by radioligand binding assays. Activation of second messenger-regulated kinases could not mimic the effect of NECA, suggesting a role for G-protein-coupled receptor kinases (GRKs). In vitro phosphorylation assays demonstrate that phosphorylation of agonist-occupied A3ARs is enhanced by GRK2 and that cellular pretreatment with NECA dramatically inhibits subsequent GRK2-mediated phosphorylation in vitro. Therefore, the A3AR is phosphorylated in situ by a kinase similar or identical to GRK2, and this may be involved in rapid functional desensitization of the A3AR. PMID- 7494006 TI - A novel heat shock response in prolactin-dependent Nb2 node lymphoma cells. AB - Virtually all cells respond to heat stress by increased expression or induction of one or more of the highly conserved cellular stress response proteins, heat shock proteins (HSPs). Here, we report the unusual property of rat Nb2-11 cells, a prolactin-dependent pre-T-cell line, to display reduced HSP expression following exposure to elevated temperature. After heat stress (41 degrees C, 1 h), there was no evidence of inducible members of the 70 kDa HSP family, a response common to other cell culture and tissue systems. Moreover, expression of constitutive members of the HSP70 and HSP90 families decreased during the heat stress, apparently reflecting a decrease in mRNA stability. Gel shift assays revealed that heat shock factor (HSF) was activated in spite of the lack of expression of inducible HSP70 transcripts, although its DNA binding rapidly deteriorated. Immunoblotting, using an antibody specific to HSF1, indicated that proteolysis of HSF1 may be responsible for this rapid termination of heat shock element binding. CCAAT binding, a component of constitutive HSP70 expression, was also reduced by heat stress in Nb2-11 cells and may account for the decline in constitutive HSP70 expression. Prolactin pretreatment prevented the fragmentation of HSF1, protected heat shock element and CCAAT binding, prevented the decline in constitutive HSP70 and HSP90 expression, and restored a modest expression of inducible HSP70 following heat treatment. Results of this study describe a unique regulatory defect in HSP expression in Nb2-11 cells, possibly a common characteristic of other hormone-dependent tumors. PMID- 7494007 TI - Interpreting chromosomal DNA restriction patterns produced by pulsed-field gel electrophoresis: criteria for bacterial strain typing. PMID- 7494008 TI - Comparison of in-house and commercially prepared haemophilus test media for disk diffusion testing of ampicillin against Haemophilus species. AB - The performance characteristics of two lots of Haemophilus Test Medium (HTM) prepared in-house at Mayo Clinic, one lot obtained from BBL (Cockeysville, Md.) and one lot obtained from Remel (Lenexa, Kans.), for disk diffusion susceptibility testing of ampicillin against 81 Haemophilus strains were assessed. Insufficient growth occurred in 2.5 to 13.6% of strains, depending on the HTM used, and was most frequent for beta-lactamase-negative, ampicillin resistant strains when they were tested with one lot of HTM prepared in-house. Results for all 14 beta-lactamase-positive Haemophilus strains were in complete agreement for all lots of HTM when either 1990 or 1993 National Committee for Clinical Laboratory Standards (NCCLS) interpretive guidelines were followed. However, these beta-lactamase-positive Haemophilus strains could be as precisely, but more rapidly, identified by the cefinase disk method. Results for 56 beta lactamase-negative, ampicillin-susceptible strains and 11 beta-lactamase negative, ampicillin-resistant strains varied significantly among all HTM lots tested and as to whether 1990 or 1993 NCCLS interpretive guidelines were followed. We conclude that in our hands disk diffusion testing with HTM prepared in-house or obtained from two commercial sources and following either 1990 or 1993 NCCLS guidelines is an unacceptable method for determining the susceptibilities of Haemophilus strains to ampicillin. PMID- 7494010 TI - Molecular method for typing Mycobacterium ulcerans. AB - A cluster of Mycobacterium ulcerans infections has recently occurred on Phillip Island, Victoria, Australia. Previous cases of infection have generally been located around Bairnsdale in southeast Gippsland. The aim of this study was to determine the epidemiological relationship between these strains and other strains originating in Australia and Africa. The previously described plasmid pTBN12 was used as a probe with restriction enzyme-digested chromosomal DNA to differentiate the strains of M. ulcerans. The probe was able to distinguish 11 restriction fragment length polymorphisms (RFLPs). Forty-three strains originating in Victoria were divided into three types, i.e., V1, V2, and V3. The majority of strains (40) yielded a type V1 pattern, including strains from southeast Gippsland. Fourteen strains from Queensland yielded three additional RFLP types, i.e., Q1, Q2, and Q3. Five strains from Benin and seven strains from Zaire yielded five additional RFLP types. It is envisaged that molecular typing of M. ulcerans strains from around the world may have a great impact on understanding of the epidemiology of infection with this organism. PMID- 7494009 TI - Corynebacterium seminale sp. nov., a new species associated with genital infections in male patients. AB - We studied 12 coryneform isolates having similar biochemical profiles which did not permit their assignment to any recognized taxa. Human semen was the source for seven of these strains, whereas the other strains were isolated from urethra, urine, and blood specimens of adult male patients. These bacteria were found in significant quantities (10(4) to 10(5) CFU/ml) in semen specimens from infertile male patients with the diagnosis of prostatitis. These strains had characteristics of the genus Corynebacterium, such as 60 mol% G + C in the DNA and corynemycolic acids, meso-diaminopimelic acid, arabinose, and galactose in the cell wall. Quantitative DNA-DNA hybridizations (S1 nuclease procedure) and phylogenies based on comparisons of almost-complete small-subunit ribosomal DNA sequences confirmed that these strains constitute a single new species within the genus Corynebacterium. All 12 strains showed similar phenotypic features, i.e., good growth on sheep blood agar in contrast with poor growth on the same medium supplemented with 1% Tween 80, a positive CAMP test in the presence of Staphylococcus aureus, glucose and sucrose fermentation, and the presence of beta glucuronidase. Some strains reduced nitrate and hydrolyzed urea or esculin. These features allowed us to distinguish these strains from members of any other coryneform taxon, and the proposed name is Corynebacterium seminale with strain IBS B12915 (CIP 104297) as the type strain. The description and delineation of these strains as a new species should be useful for further studies, including evaluations of their prevalence among the normal flora and their clinical implications. PMID- 7494011 TI - Evaluation of mupirocin E-test for determination of isolate susceptibility: comparison with standard agar dilution techniques. AB - Mupirocin E-test strips have been evaluated for their ease of use and accuracy in determining the susceptibilities of 171 strains of Staphylococcus spp., Streptococcus spp., Haemophilus influenzae, and Moraxella catarrhalis. The susceptibility of each strain was determined on two occasions, using parallel E test and agar dilution methodologies each time. To ensure similar precisions for statistical analyses, E-test MICs were rounded up to a standard twofold agar dilution scale. Clear, elliptical zones were obtained against Staphylococcus spp. M. catarrhalis also gave clear zones, but the scale intercept was often difficult to interpret because of the irregular shape of the inhibition zone. Poor growth sometimes resulted in less-distinct zones of inhibition against Streptococcus spp. and H. influenzae. Excellent correlation was observed between the the E-test and agar dilution against Staphylococcus spp. and H. influenzae, with > 95% of the E-test values falling within one log2 dilution of the corresponding agar MIC. The correlation was lower for Streptococcus spp. and M. catarrhalis, with 86 and 83%, respectively, of E-test results falling within one log2 dilution of the agar MIC. When E-test MICs did not agree exactly with the corresponding agar MIC against Staphylococcus spp. or Streptococcus spp., there was a tendency for the E test to give a lower MIC. This bias has little effect upon individual MICs in staphylococci or in the generation of susceptibility interpretation errors ( < 1.5% overall), but it could reduce population geometric mean MICs by factors of 0.78 to 0.83. This effect was more marked for Streptococcus spp., reducing the population mean by a factor of 0.73 and resulting in 0.7% major and 8% very major errors. In contrast, the E-test tended to give higher MICs against M. catarrhalis, resulting in 7.3% major errors and increasing the population geometric mean MIC by a factor of 1.60. PMID- 7494012 TI - Antibodies against whole sonicated Borrelia burgdorferi spirochetes, 41 kilodalton flagellin, and P39 protein in patients with PCR- or culture-proven late Lyme borreliosis. AB - The sensitivities and specificities of three enzyme-linked immunosorbent assays (ELISAs) for Borrelia burgdorferi antibodies were compared for 41 patients presenting with symptoms compatible with late Lyme borreliosis (LB) and 37 healthy controls. All subjects were living in southwestern Finland, where LB is endemic. Only patients with culture- or PCR-proven disease were enrolled in the study. The antigens of the ELISAs consisted of sonicated spirochetes, 41-kDa flagellin, and recombinant P39 protein of B. burgdorferi. Fifteen patients had strongly or moderately positive results in the serological assay(s), 19 patients had only weakly positive or borderline antibody levels, and the remaining 7 patients were seronegative by ELISA. The sensitivities of the ELISAs were 78.0% with sonicate antigen, 41.5% with 41-kDa flagellin, and 14.6% with P39 protein. The specificities of the tests were 89.2, 86.5, and 94.6%, respectively. The sonicate antigen ELISA seems to be an effective screening method. These results show that antibodies to B. burgdorferi may be present in low levels or even absent in patients with culture- or PCR-proven late LB. Therefore, in addition to serological testing, the use of PCR and cultivation is recommended in the diagnosis of LB. PMID- 7494013 TI - Controlled evaluation of BacT/alert standard anaerobic and FAN anaerobic blood culture bottles for the detection of bacteremia and fungemia. AB - FAN medium was formulated to improve microbial recovery, particularly for fastidious microorganisms and for microorganisms causing sepsis in patients receiving antimicrobial therapy. In a controlled clinical evaluation performed at four university-affiliated hospitals, FAN anaerobic bottles were compared with standard anaerobic bottles for yield, speed of detection of microbial growth, and detection of septic episodes. A total of 10,431 blood culture sets were received; both anaerobic bottles of 7,694 blood culture sets were adequately filled with blood. Altogether, 925 isolates were recovered: 557 that were the cause of sepsis, 99 that were indeterminate as the cause of sepsis, and 269 contaminants. More Staphylococcus aureus (P < 0.001), coagulase-negative staphylococci (P < 0.001), Escherichia coli (P < 0.02), and all microorganisms combined (P < 0.005) were recovered from FAN bottles; more nonfermentative gram-negative bacilli (P < 0.05), Torulopsis glabrata (P < 0.001), and other yeasts (P < 0.01) were recovered from standard bottles. Growth of S. aureus (P < 0.001), coagulase negative staphylococci (P < 0.001), Enterococcus faecalis (P < 0.025), streptococci other than Streptococcus pneumoniae (P < 0.01), and all microorganisms combined (P < 0.001) was detected earlier in standard bottles; growth of more isolates of E. coli (P < 0.05) and anaerobic bacteria (P < 0.01) was detected earlier in FAN bottles. The mean times to detection were 14.2 and 16.1 h for standard and FAN bottles, respectively. More septic episodes caused by S. aureus (P < 0.001), coagulase-negative staphylococci (P < 0.005), members of the family Enterobacteriaceae (P < 0.02), and all microorganisms combined (P < 0.02) were detected in FAN bottles; more septic episodes caused by nonfermentative gram-negative bacilli (P < 0.025) and yeasts (P < 0.005) were detected in standard bottles. In summary, more isolates (except for strict aerobes) were recovered from FAN bottles than from standard anaerobic bottles. Similarly, significant more septic episodes (except for those caused by strict aerobes) were detected with FAN bottles than with standard anaerobic bottles. With the exception of E. coli and anaerobic bacteria, growth of more isolates was detected earlier in standard anaerobic bottles. PMID- 7494014 TI - Comparison of Mycoplasma arthritidis strains by enzyme-linked immunosorbent assay, immunoblotting, and DNA restriction analysis. AB - Twenty Mycoplasma arthritidis strains or isolates were compared by a combination of enzyme-linked immunosorbent assay by an antiserum adsorption technique, Western immunoblotting, and restriction analysis of chromosomal DNA. Antigenic markers that defined strains related to strains 158p10p9, PG6, and H606 were identified. In addition, restriction analysis allowed all 20 strains to be divided into six groups. Results of restriction analysis corresponded generally with antigenic similarities, although the former did not allow grouping with as fine a precision as the latter. However, intrastrain antigenic variability, which is common among many Mycoplasma species, including M. arthritidis, introduced a complicating factor into our attempts at antigenic analysis. While serologic and antigenic analyses remain useful, we recommend that they be used with caution and in combination with other techniques for identifying and characterizing new isolates and newly acquired strains. Combinations of these techniques have proven to be useful in our laboratory for quality control and for uncovering interesting relationships among strains subjected to animal passage and their less virulent antecedents and among strains originally classified as the same but obtained from different sources and maintained, sometimes for decades, in different laboratories. PMID- 7494016 TI - Demonstration of homology between IS6110 of Mycobacterium tuberculosis and DNAs of other Mycobacterium spp.? AB - The insertion sequence IS6110 has an important role in diagnostic PCR and typing of Mycobacterium tuberculosis. We have evaluated a one-tube nested PCR which detects IS6110. Positive results were obtained with DNAs from four of four M. tuberculosis isolates, seven of eight M. fortuitum isolates, seven of seven M. avium-M. intracellulare complex isolates, four of five M. kansasii isolates, four of five M. xenopi isolates, two of four M. malmoense isolates, and one of two M. chelonei isolates. These results were confirmed by hybridization of genomic DNA from bp 505 to 685 of the IS6110 from M. tuberculosis H37Rv. Dot blot hybridization of genomic DNAs from these isolates with the same probe cinfirmed the presence of a homologous sequence in these mycobacterial species. These data suggest that false-positive results may be obtained for clinical samples when some methods based on IS6110 are used [corrected]. PMID- 7494015 TI - Characterization of feline Helicobacter pylori strains and associated gastritis in a colony of domestic cats. AB - Twenty-four young adult domestic cats from a commercial vendor were found to be infected with Helicobacter pylori. Histopathologic analyses, selected electron microscopy, and urease mapping were performed on mucosal samples collected from the cardias and fundi, bodies, and antra of these cats' stomachs. H. pylori organisms were abundant in all areas of the stomach on the basis of histologic evaluation and urease mapping. H. pylori infection was associated with a moderate to severe lymphofollicular gastritis in 21 of 24 cats (88%). The gastritis was most pronounced in the antral region and consisted mainly of multifocal lymphoplasmacytic follicular infiltrates in the deep mucosa. The severity of gastritis in the antrum corresponded to high numbers of H. pylori there on the basis of the use of the urease assay as an indicator of H. pylori colonization. Ten of 24 cats (42%) also had small to moderate numbers of eosinophils in the gastric mucosa. All 24 cats had gastric lymphoid follicles, with follicles being most prevalent in the antrum. Electron microscopy of gastric tissue revealed numerous H. pylori organisms, some of which were closely adhered to the mucosal epithelium. Human H. pylori gene-specific primers to ureA and ureB amplified products of similar sizes from H. pylori cat isolates. Digestion of the products with restriction enzymes resulted in fragments characteristic of the restriction fragment length polymorphism patterns of H. pylori isolates from humans. In the domestic cat, H. pylori infection is associated with a lymphofollicular gastritis, consisting of lymphocytic and plasmacytic infiltration into the lamina propria, and the organism appears to provide chronic antigenic stimulation resulting in the formation of gastric lymphoid follicles. PMID- 7494017 TI - Detection of microsporidia (Enterocytozoon bieneusi) in intestinal biopsy specimens from human immunodeficiency virus-infected patients by PCR. AB - Intestinal microsporidiosis has been implicated as a major cause of chronic diarrhea in human immunodeficiency virus (HIV)-infected patients. So far diagnosis depends on direct visualization of the parasites by light and transmission electron microscopy. We evaluated the diagnostic value of microsporidian DNA amplification by PCR on duodenal biopsy specimens obtained from patients with and without intestinal microsporidiosis caused by Enterocytozoon bieneusi. Thirteen HIV-infected patients (all CDC stage C3) were studied. Eight patients had intestinal microsporidiosis caused by E. bieneusi (n = 6), Septata intestinalis (n = 1), and Encephalitozoon cuniculi (n = 1); microsporidioses were diagnosed by light microscopy of stool samples and confirmed by light and electron microscopy of intestinal biopsy specimens. Five patients had no microsporidia in their stool samples or in their intestinal biopsy specimens, as examined by light and electron microscopy. Additionally, DNA prepared from Toxoplasma gondii derived from mouse ascites was used as a further control. A 353-bp DNA fragment of the small-subunit rRNA gene could be amplified from all six biopsy specimens infected with E. bieneusi, and the nature of the PCR products was confirmed by Southern blot hybridization. No amplification of DNA fragments was seen by using DNA extracted from biopsy specimens with S. intestinalis or E. cuniculi infection or without microsporidian infection and with template DNA extracted from T. gondii. The results suggest that PCR testing of intestinal biopsy specimens may be a useful approach to diagnosing microsporidiosis in HIV-infected patients. PMID- 7494018 TI - Simplified procedure for preparation of sensitized latex particles to detect capsular polysaccharides: application to typing and diagnosis of Actinobacillus pleuropneumoniae. AB - A novel, inexpensive method for obtaining immunoglobulin G (IgG) specific for capsular antigen is described for use in latex agglutination tests. Hyperimmune rabbit serum against encapsulated Actinobacillus pleuropneumoniae was thoroughly adsorbed with a nonencapsulated mutant. The capsule titer of the absorbed serum was unaffected, whereas reactivity to nonencapsulated cells was reduced to background levels, as determined by enzyme immunoassay. The IgG component of the adsorbed serum was recovered by protein A chromatography and was covalently coupled through a water-soluble carbodiimide to carboxylate latex beads. The sensitized latex particles (SLP) were agglutinated by 10 ng of homologous capsule or more per ml, were not agglutinated by heterologous capsules at concentrations of < 10 micrograms/ml, and were stable for over 1 year at 4 degrees C without loss of sensitivity. There was no difference in the sensitivity or specificity of latex particles coupled with IgG purified by capsule affinity chromatography. The SLP were agglutinated by all strains of bacteria of the homologous serotype but not by heterologous serotypes or strains of Pasteurella multocida, Actinobacillus suis, or Haemophilus parasuis tested at a density equivalent to a 0.5 McFarland standard. The SLP detected homologous capsule in lung tissue, nasal swabs, and concentrated urine samples from all pigs culture positive for A. pleuropneumoniae but one. Precoating of carboxylate latex particles with avidin followed by conjugation of biotin-hydrazide-labelled IgG to capsule increased the sensitivity of the assay approximately 10-fold. Adsorption of serum with nonencapsulated mutants may be used to prepare SLP with optimum sensitivity and specificity without the need to purify capsule or couple capsule to affinity columns. PMID- 7494020 TI - Direct antimicrobial susceptibility testing for acute urinary tract infections in women. AB - Despite its theoretical advantages, direct antimicrobial susceptibility testing (DST) of urine specimens remains controversial largely because of concerns regarding its accuracy, particularly with mixed cultures. To evaluate the performance of DST in the setting of acute urinary tract infection (UTI), we performed DST using 25 traditional and contemporary antimicrobial agents on urine specimens from 162 women with suspected acute uncomplicated UTI, and compared these results with the results of standardized disk diffusion susceptibility tests done on the same specimens. Direct tests were interpretable for 129 specimens, i.e., 80% of all specimens and 85% of the 152 specimens that met the culture criteria for UTI. Of the 2,983 individual comparisons between the direct and standard tests, 0.8% represented very major errors, 0.6% represented major errors, 3.1% represented minor errors, and 95.5% were in agreement. Errors were more common in association with older antimicrobial agents and agents with a high prevalence of antimicrobial resistance, non-Escherichia coli strains, low urine bacterial concentrations, sparse or mixed growth in the direct test, and the presence of multiple significant organisms in urine. The urine leukocyte concentration was > or = 15/mm3 in all subjects and did not differentiate between specimens that gave an interpretable direct test and those that did not. Calculation of the sensitivity of DST in identifying antimicrobial resistance supplemented conventional error rate analysis. We conclude that when used selectively and interpreted carefully, DST of urine specimens offers an efficient, rapid, and accurate method for antimicrobial susceptibility determination for acute UTI, particularly when the urine bacterial concentration is > 10(5) CFU/ml. PMID- 7494021 TI - Colorimetric method for determining MICs of antimicrobial agents for Mycobacterium tuberculosis. AB - A colorimetric method for quantitative measurement of the susceptibility of Mycobacterium tuberculosis to antimicrobial agents is described. The method utilizes an oxidation-reduction dye, Alamar blue, as an indicator of growth. By this method, MICs of isoniazid, rifampin, streptomycin, and ethambutol were determined for 50 strains of M. tuberculosis. Colorimetric MIC results were available on the 7th, 10th, or 14th day of incubation for 29 (58%), 14 (28%), and 7 (14%) of the 50 strains, respectively. When MIC susceptibility results were compared with results obtained by the agar proportion method, increased levels of resistance detected by agar proportion were associated with higher MICs obtained by the colorimetric method. Tentative interpretive criteria for colorimetric MIC results which showed good agreement with results obtained by the agar proportion method were established. Interpretive agreement between the two methods was 98% for isoniazid, rifampin, and ethambutol and 94% for streptomycin. Overall, there was agreement between the two methods for 194 of 200 test results (97%). The colorimetric method is a rapid, quantitative, nonradiometric method for determining the antimicrobial susceptibility of M. tuberculosis. PMID- 7494019 TI - Genotypic and phenotypic characterization of Borrelia burgdorferi isolated from ticks and small animals in Illinois. AB - We have characterized 33 isolates of Borrelia burgdorferi from northern Illinois (32 isolates) and Wisconsin (1 isolate) representing the largest series of midwestern isolates investigated to date. The techniques used for molecular analysis of strains included (i) genospecies typing with species-specific PCR primers, (ii) plasmid profiling by pulsed-field gel electrophoresis of total genomic DNA, (iii) large-restriction-fragment pattern (LRFP) analysis by pulsed field gel electrophoresis of MluI-digested genomic DNA (J. Belfaiza, D. Postic, E. Bellenger, G. Baranton, and I. Saint Girons, J. Clin. Microbiol. 31:2873-2877, 1993), (iv) sodium dodecyl sulfate-polyacrylamide gel electrophoresis of total proteins, (v) microsequencing of high-performance liquid chromatography-purified peptides derived from proteins showing high levels of expression, (vi) amino acid composition analysis of proteins, and (vii) immunological analysis of proteins with a polyclonal antiserum of human origin. Five reference strains as well as two atypical tick isolates from California (DN127) and New York (25015) were included for comparison. All of the Illinois and Wisconsin isolates were typed as B. burgdorferi sensu stricto with genospecies-specific PCR primers. The isolates were found to be heterogeneous with regard to their plasmid and protein profiles. One isolate from Illinois possessed two large-molecular-size plasmids instead of the usual 49-kb plasmid. Fragment patterns resulting from MluI digestion of genomic DNA from the 33 isolates and strains DN127 and 25015 were separable into six distinct LRFPs, five of which have not previously been described. Strain 25015 and an isolate from Illinois (CT39) shared an unusual LRFP that is not typical of other B. burgdorferi sensu stricto strains, suggesting that they may represent a fifth species of B. burgdorferi sensu lato. Five of the 33 isolates and strains DN127 and 25015 showed high-level expression of proteins with molecular masses of approximately 22 kDa. Investigation of these proteins by microsequencing of individual peptides and total amino acid composition analysis indicated that the 22-kDa proteins expressed by the seven strains were polymorphic OspC proteins. By using a polyclonal serum of human origin, expression of OspC could be detected in all 33 Illinois and Wisconsin isolates. PMID- 7494022 TI - Comparison of in vitro culture, immunohistochemical staining, and PCR for detection of Borrelia burgdorferi in tissue from experimentally infected animals. AB - An avidin-biotin-amplified immunophosphatase staining method with a purified polyclonal rabbit anti-Borrelia burgdorferi hyperimmune serum was developed for identification of B. burgdorferi in tissue specimens. The diagnostic efficacy was compared with those of in vitro culture and PCR with fresh and fixed, paraffin embedded tissues. A nested PCR assay was developed for identification of a 276-bp fragment of the B. burgdorferi flagellin gene. The diagnostic sensitivities of the different techniques were evaluated with spleen, renal, and urinary bladder tissues from eight experimentally infected gerbils. A systemic infection was verified by positivity of 23 of 24 (96%) organ cultures. B. burgdorferi was visualized immunohistochemically in 9 of 23 (39%) of the specimens. Among these nine specimens, an average of 33% of the 15 sections examined were positive. The spirochetes accumulated in discrete clusters and were associated with focal lymphocytic infiltration. The diagnostic sensitivity obtained by PCR with fixed, paraffin-embedded tissue was 21%, considerably lower than that with fresh tissue (71%). Thus, the reliable demonstration of B. burgdorferi by immunohistochemical staining is possible but extremely laborious, and considering the fact that the density of B. burgdorferi in human tissue is even lower than that in experimentally infected animals, the method is not useful in a clinical setting. It may, however, still be valuable in pathogenetic research. Detection of B. burgdorferi DNA by PCR should be performed with fresh tissue specimens and not with fixed, paraffin-embedded specimens. PMID- 7494023 TI - Evaluation of accuracy and reproducibility of E test for susceptibility testing of Streptococcus pneumoniae to penicillin, cefotaxime, and ceftriaxone. AB - We evaluated the reproducibility with which technologists perform and interpret the E test (AB Biodisk, North America, Inc., Piscataway, N.J.) for determining the susceptibility of Streptococcus pneumoniae to penicillin, cefotaxime, and ceftriaxone. Four technologists prepared E test assays to test 124 isolates of S. pneumoniae. Each technologist then interpreted the results of the E test blinded to the interpretation of the other technologists. In addition, E test results were compared with the reference method of broth microdilution. Intraobserver and interobserver agreement were assessed by use of the kappa statistic. Interpretation of the E test and broth microdilution results showed substantial to excellent agreement, with kappa values ranging from 0.878 to 0.987. Compared with broth microdilution, no very major errors and only four major errors were made with the E test. Most minor errors with penicillin and ceftriaxone occurred for isolates with intermediate or high-level resistance, whereas for cefotaxime the minor errors were more evenly distributed between susceptible and intermediate resistance and between intermediate and high-level resistance. These results indicate that there is good agreement between technologists for the interpretation of the E test when testing the susceptibility of S. pneumoniae to penicillin, cefotaxime, and ceftriaxone and that the results of the E test agree with those of broth microdilution. PMID- 7494024 TI - Detection of human immunodeficiency virus (HIV) type 1 antibodies by new automated microparticle enzyme immunoassay for HIV types 1 and 2. AB - We compared an automated microparticle double-antigen sandwich enzyme immunoassay (EIA) for the IMx test system recently developed by Abbott with two established assays (the automated indirect Vidas IgG EIA and the double-antigen sandwich EIA from Murex/Wellcome) devised for the detection of human immunodeficiency virus type 1 (HIV-1) and HIV-2 antibodies. A total of 1,078 consecutive serum samples were tested prospectively with the three assays. In addition, we used retrospectively selected panels of serum samples with discrepant results in two different screening tests and with indeterminate or positive Western immunoblot (WB) results, as well as five commercially available HIV-1 seroconversion panels. The new assay showed excellent discriminatory characteristics for the separation of samples from HIV-1-positive and HIV-1-negative persons according to Centers for Disease Control and Prevention WB criteria. The sensitivities were 98.1, 92.9, and 96.1% for the new test and the two other assays, respectively, and the specificities were 99.7, 97.9, and 98.1%, respectively. With the seroconversion panels this new test was positive several days earlier than the two other assays; i.e., seroconversion was evident at the peak of p24 antigenemia and often several weeks before WB became positive by the most stringent criteria. PMID- 7494025 TI - Species-specific PCR detection of malaria parasites by microtiter plate hybridization: clinical study with malaria patients. AB - A simple and convenient PCR method that amplifies the 18S rRNA genes has been developed for the purpose of detecting and differentiating four species causing malaria in humans. The advantage of the assay is that the biotinylated PCR product is visualized following hybridization with specific probes which are immobilized on plate wells (microtiter plate hybridization). This method has been previously evaluated in a field study and was found to be sensitive and specific for the detection of Plasmodium falciparum and Plasmodium vivax. In the current study, the microtiter plate hybridization PCR method was evaluated by using blood specimens from malaria patients. All of 36 cases of falciparum malaria, 26 of 27 cases of vivax malaria, all of 11 cases of ovale malaria, and 2 cases of malariae malaria were diagnosed species specifically by the PCR method. There were four smear-negative, PCR-positive cases that seemed to correspond to the convalescent stage of malaria. In contrast, 30 cases for which the diagnosis of malaria has been excluded on the basis of microscopy and clinical courses showed negative PCR results. By comparing parasite densities and PCR results following antimalarial treatment of some patients, it was revealed that the PCR results largely paralleled the parasite densities and that PCR could detect as few as 10 parasites per microliter of blood. We conclude that this PCR method is highly sensitive and specific for the detection of all four parasite species and can serve as a useful supplement to microscopy for the clinical management of malaria. PMID- 7494026 TI - Incidence of the highly conserved fib gene and expression of the fibrinogen binding (Fib) protein among clinical isolates of Staphylococcus aureus. AB - We have recently described a 19-kDa fibrinogen-binding protein, termed Fib, which is secreted into the extracellular medium by Staphylococcus aureus. In this study, the presence of the Fib protein and the fib gene among clinical isolates of S. aureus and among other staphylococcal species known to interact with fibrinogen was investigated. This task was pursued at the DNA, mRNA, and protein levels. It was found that the fib gene was unique to S. aureus and highly conserved at the nucleotide level. The Fib protein was produced by all S. aureus strains investigated but was not detected in all bovine mastitis strains, because of proteolytic degradation by simultaneously produced staphylococcal proteases. It was concluded that the fib gene was unique to S. aureus and that it could be used in the identification of S. aureus. PMID- 7494027 TI - Analysis of strains of Burkholderia (Pseudomonas) cepacia isolated in a nosocomial outbreak by biochemical and genomic typing. AB - We analyzed strains of Burkholderia (formerly Pseudomonas) cepacia isolated in a nosocomial outbreak by biochemical and genomic typing methods. One hundred isolates of B. cepacia were obtained from patients at several wards in a single hospital from March 1983 to February 1984. These isolates were classified into 12 groups by a new biochemical typing scheme on the basis of the production of six enzymes and the production of hemolytic substance and yellow pigment. Among them, 33 strains collected from the 12 groups were further characterized by DNA fingerprinting by pulse-field gel electrophoresis, ribotyping, and plasmid profile analysis. Forty-seven strains of B. cepacia of independent origins and 25 isolates from the same hospital obtained 10 years later for a follow-up study were also subjected to analysis. Both DNA fingerprinting and ribotyping clearly discriminated the isolates from different hospitals. Of interest, although the biochemical typing and plasmid profiles of the isolates obtained during 1983 to 1984 in a single hospital were variable, genomic typing identified the majority of the isolates (32 of 33 [97%]) as derivatives of a single strain. Furthermore, a follow-up study suggested the persistence of such derivatives among the isolates after a decade. These results clearly indicated that the outbreak of B. cepacia infection in the hospital was nosocomial in origin. Thus, the usefulness of genomic typing for epidemiological studies of B. cepacia infection was further demonstrated. The biochemical typing revealed the marked variability of phenotypes of B. cepacia. PMID- 7494028 TI - Characterization of a 17-kilodalton antigen of Bartonella henselae reactive with sera from patients with cat scratch disease. AB - A library of Bartonella (Rochalimaea) henselae DNA was constructed in the cloning vector lambda ZAPII and screened for expression of antigenic proteins by using a pool of sera from patients who had been diagnosed with cat scratch disease (CSD) and had antibodies to Bartonella spp., as determined by indirect fluorescent antibody (IFA) assay. Ten immunoreactive phages were subcloned as recombinant plasmids by in vivo excision. All 10 recombinants expressed a protein of approximately 17 kDa when they were examined by immunoblot with the pool of human sera. Restriction endonuclease digestion of each recombinant plasmid indicated seven profiles, suggesting that cloning bias was not the reason for repeated isolation of clones expressing the 17-kDa antigen. The gene coding for the 17-kDa antigen was sequenced and shown to code for an open reading frame of 148 amino acids with a predicted molecular mass of 16,893 Da. The amino terminus of the deduced amino acid sequence was hydrophobic in nature and similar in size and composition to signal peptides found in gram-negative bacteria. The remainder of the deduced amino acid sequence was more hydrophilic and may represent surface exposed epitopes. Further subcloning of the 17-kDa antigen as a biotinylated fusion protein in the expression vector PinPoint Xa-2 resulted in a 30-kDa protein that was highly reactive on immunoblots with individual serum samples from patients with CSD. The agreement between reactivity with the 30-kDa fusion protein on immunoblot analysis and the results obtained by IFA assay was 92% for IFA-positive sera and 88% for IFA-negative sera. The recombinant-expressed 17-kDa protein should be of value as an antigen for serologic diagnosis of CSD and Bartonella infections and warrants further study in attempts to develop a subunit vaccine to prevent long-term Bartonella infection in cats and the potential for further spread of these organisms to humans. PMID- 7494029 TI - Use of random amplified polymorphic DNA as a typing method for Candida albicans in epidemiological surveillance of a burn unit. AB - Burn patients are particularly exposed to deep-seated nosocomial infections caused by Candida species. Superficial carriage of C. albicans is a potential source of infection and dissemination, and typing methods could be useful to trace the different isolates. We report the use of random amplified polymorphic DNA to type isolates of C. albicans in the Hopital Cochin burn unit. This molecular typing method, which is based on PCR with arbitrary short primers, was evaluated on a panel of 32 C. albicans strains isolated from various anatomical sites of unrelated patients, and the strains showed 22 different patterns. Random amplified polymorphic DNA was then used in the epidemiological surveillance of the patients in the burn unit over a 9-month period. Seven patterns were identified among 84 isolates from 18 patients. One pattern (pattern A) corresponding to isolates from 7 of the 18 patients (68% of isolates) predominated throughout the 9-month study, while some strains with other profiles were isolated only once. Some profiles appeared to show a particular geographic pattern within the unit, suggesting transmission from room to room. These results underline the importance of fungal surveillance in such patients and the need to inform nursing staff of measures to prevent the spread of Candida spp. from patient to patient. PMID- 7494030 TI - Characterization of Hafnia alvei by biochemical tests, random amplified polymorphic DNA PCR, and partial sequencing of 16S rRNA gene. AB - Hafnia alvei strains which possess the attachment-effacement gene (eaeA) may have clinical importance as new diarrhea-causing pathogens and should therefore be differentiated from other H. alvei strains. We characterized diarrheal H. alvei strains, which were positive in the PCR test for the eaeA gene, using biochemical tests not routinely used for identification of members of the family Enterobacteriaceae, and compared them with eaeA-negative strains isolated from different clinical and nonclinical sources to find characteristics useful for identification. Random amplified polymorphic DNA (RAPD)-PCR and partial sequencing of the 16S rRNA gene were utilized to study the genetic diversity of the isolates. The eaeA-positive strains were found to have many characteristic biochemical properties. Negative reactions in the 2-ketogluconate and histidine assimilation tests and a positive reaction in the 3-hydroxybenzoate assimilation test may be useful in routine diagnostics. Nearly identical RAPD-PCR profiles and identical 353-bp fragments of the 16S rRNA genes indicated little genetic diversity among the eaeA-positive strains. The low level of homology (92%) in the partial 16S rRNA genes of eaeA-positive and -negative H. alvei strains raises questions about the taxonomic positioning of eaeA-positive H. alvei. PMID- 7494032 TI - Clinical and microbiological features of necrotizing fasciitis. AB - The microbiological and clinical characteristics of 83 patients with necrotizing fasciitis (NF) treated over a period of 17 years are presented. Bacterial growth was noted in 81 of 83 (98%) of specimens from patients with NF. Aerobic or facultative bacteria only were recovered in 8 (10%) specimens, anaerobic bacteria only were recovered in 18 (22%) specimens, and mixed-aerobic-anaerobic floras were recovered in 55 (68%) specimens. In total, there were 375 isolates, 105 aerobic or facultative bacteria and 270 anaerobic bacteria, for an average of 4.6 isolates per specimen. The recovery of certain bacteria from different anatomical locations correlated with their distribution in the normal flora adjacent to the infected site. Anaerobic bacteria outnumbered aerobic bacteria at all body sites, but the highest recovery rate of anaerobes was in the buttocks, trunk, neck, external genitalia, and inguinal areas. The predominant aerobes were Staphylococcus aureus (n = 14 isolates), Escherichia coli (n = 12), and group A streptococci (n = 8). The predominant anaerobes were Peptostreptococcus spp. (n = 101), Prevotella and Porphyromonas spp. (n = 40), Bacteroides fragilis group (n = 36), and Clostridium spp. (n = 23). Certain clinical findings correlated with some bacteria: edema with B. fragilis group, Clostridium spp., S. aureus, Prevotella spp. and group A streptococci; gas and crepitation in tissues with members of the family Enterobacteriaceae and Clostridium spp.; and foul odor with Bacteroides spp. Certain predisposing conditions correlated with some organisms: trauma with Clostridium spp.; diabetes with Bacteroides spp., members of the family Enterobacteriaceae, and S. aureus; and immunosuppression and malignancy with Pseudomonas spp. and members of the family Enterobacteriaceae. These data highlight the polymicrobial nature of NF. PMID- 7494031 TI - Intergenic 16S rRNA gene (rDNA)-23S rDNA sequence length polymorphisms in members of the family Legionellaceae. AB - A method based on PCR amplification of the 16S rRNA gene (rDNA)-23S rDNA intergenic regions was developed for the identification of species within the family Legionellaceae. The sizes of the PCR products varied from 1,353 to 350 bp. Strains of Legionella pneumophila were characterized as having products of approximately 900 and 530 bp, and L. birminghamensis had products of 1,390, 960, and 380 bp. Of the 38 species of legionellae examined, only 7 were indistinguishable (L. erythra from L. rubrilucens, L. anisa or L. cherrii from L. tucsonensis, and L. quateirensis from L. shakespearei). Two environmental isolates were identified as L. pneumophila. Strain LLAP-3, which was a symbiont of amoebae, could not be associated with any Legionella sp. studied. PMID- 7494033 TI - Evaluation of AnaeroGen system for growth of anaerobic bacteria. AB - The Oxoid AnaeroGen system was compared with the BBL GasPak for the production of an anaerobic atmosphere and was evaluated for its ability to support the growth of 135 clinically significant anaerobic bacteria. An anaerobe chamber was used as the "gold standard" for supporting the growth of anaerobes. The AnaeroGen requires no catalyst, produces no hydrogen, requires no water, and reduces preparation time to a minimum. The water-activated BBL GasPak generates hydrogen. For 132 of the 135 strains tested, better initial growth at 48 h was noted for the jar methods than for the anaerobe chamber. At 72 h, 113 of the 135 strains showed equal growth, and at 7 days, only marginal differences in growth patterns were noted. The AnaeroGen never failed to reduce the anaerobic indicator, while the BBL GasPak occasionally failed to do so. The AnaeroGen performed at least as well as, and sometimes better than, the established methods. The AnaeroGen is a good alternative for use in anaerobic jars. PMID- 7494034 TI - Evaluation of Cobas Core Rubella IgG EIA recomb, a new enzyme immunoassay based on recombinant rubella-like particles. AB - Cobas Core Rubella IgG EIA recomb (Roche), a new, commercially available rubella enzyme immunoassay using recombinant rubella-like particles, was compared with a hemagglutination inhibition assay (Rubenosticon; Organon Teknika) and two whole virus enzyme immunoassays (IMx [Abbott] and Platelia [Sanofi Diagnostics Pasteur]). Compared with those of these reference assays, the relative sensitivities of Cobas Core Rubella IgG recomb were 100, 94, and 95.9%, with specificities amounting to 80.8, 98, and 98.2%, respectively. PMID- 7494035 TI - Evaluation of Rapid ATB Staph for 5-hour antimicrobial susceptibility testing of Staphylococcus aureus. Groupement pour le Depistage, L'Etude et la Prevention des Infections Hospitalieres-Groep ter Opsporing, Studie en Preventie van Infecties in de Ziekenhuizen. AB - The accuracy of Rapid ATB Staph (bioMerieux, La Balme-Les Grottes, France) for detection of oxacillin resistance and for detection susceptibility to 11 other antimicrobial agents in 553 and 519 Staphylococcus aureus isolates, respectively, was evaluated by comparing results with those produced by oxacillin agar screen and agar dilution methods, respectively. Further characterization of isolates with discrepant results for oxacillin testing was done by PCR detection of the nuc and mecA genes. By oxacillin agar screening, there were 307 oxacillin resistant and 246 oxacillin-susceptible isolates. Rapid ATB results were obtained in 5 h for 515 (93.2%) of the isolates tested. Rapid ATB showed 97.0% sensitivity for detection of oxacillin resistance, confirmed by the presence of the mecA gene. After repeat testing of isolates flagged by the ATB software as possible errors, sensitivity increased to 99% for oxacillin-resistant isolates. Essential agreement with agar dilution testing for susceptibility to amoxicillin-clavulanic acid, gentamicin, erythromycin, clindamycin, and ciprofloxacin, as estimated by Youden's J statistic, was > 0.90. Subpopulations of isolates with significantly increased MICs of amikacin, rifampin, and minocycline, indicating borderline susceptibility, were detected by Rapid ATB and categorized as resistant. Rapid ATB Staph showed adequate accuracy for detection within 5 h of the oxacillin- and multiple-drug-resistant S. aureus isolates currently prevalent in Belgium. PMID- 7494036 TI - Geographic spread of epidemic multiresistant Staphylococcus aureus clone in Brazil. AB - Staphylococcus aureus isolates from five large teaching hospitals and one medium size community hospital located in geographically distant parts of Brazil, in the south and southeast (Rio de Janeiro, Niteroi, Sao Paulo, Porto Alegre) and in the north (Manaus), were tested for their antibiotic resistance patterns and genetic backgrounds. Eighty-five of the 152 isolates were identified as methicillin resistant S. aureus (MRSA) by using a combination of an agar dilution screen and a mecA gene-specific DNA probe. All MRSA isolates were resistant to penicillin, erythromycin, gentamicin, oxacillin, and cephalothin, and the majority of isolates (74%) were also resistant to chloramphenicol, sulfamethoxazole trimethoprim, ciprofloxacin, and clindamycin as well and were susceptible only to vancomycin. Isolates obtained from hospitals in Sao Paulo, Rio de Janeiro, Niteroi, and Porto Alegre (1,600 km from one another) and Manaus (3,700 km from Rio de Janeiro) were examined by a variety of molecular fingerprinting techniques: the nature of the mecA polymorph and Tn554 attachment sites and restriction fragment length polymorphism of genomic DNAs after SmaI restriction and separation of the digested DNA by pulsed-field gel electrophoresis. The overwhelming majority of the isolates shared a common pulsed-field gel electrophoresis pattern and carried mecA polymorph III in combination with Tn554 pattern B, indicating the presence of a single, epidemic MRSA clone spread over large geographic distances of Brazil. PMID- 7494038 TI - In vitro susceptibility of human vascular wall cells to infection with Chlamydia pneumoniae. AB - Chlamydia pneumoniae is a common respiratory pathogen. Recent studies have demonstrated the presence of C. pneumoniae in coronary and aortic atherosclerotic lesions. To study the role of C. pneumoniae in atherosclerosis, we investigated the susceptibilities of three different cells of the human vascular wall to infection with C. pneumoniae AR-39. These cell types were endothelial cells, smooth muscle cells, and macrophages derived from peripheral blood monocytes. Infection was assessed by using a direct fluorescent antibody to assess inclusion counts. Duplicate cell samples were harvested 3 days postinfection and were passed in HL cells, a susceptible human epithelial cell line, to determine if infectious organisms were produced. Endothelial cells, smooth muscle cells, and macrophages were capable of supporting C. pneumoniae growth in vitro. These results showed that three different cell types known to be important in atherogenesis are susceptible to infection with C. pneumoniae. PMID- 7494039 TI - Outer surface protein C gene sequence analysis of Borrelia burgdorferi sensu lato isolates from Japan. AB - The nucleotide sequences of the outer surface protein C gene (ospC) from Borrelia burgdorferi sensu lato isolates representing six different restriction fragment length polymorphism (RFLP) ribotype groups were determined, and the deduced amino acid sequences were aligned in comparison with the previously published OspC protein sequences. The sequence similarity analysis revealed the high sequence variability of OspC protein, and the degree of amino acid similarity ranged from 53.8 to 100% among 25 isolates. It has been reported that the representatives belonging to the three species of B. burgdorferi sensu lato showed a species specific amino acid sequence motif at positions 23 to 35 (B. Wilske, S. Jauris Heipke, R. Lobentanzer, I. Pradel, V. Preac-Mursic, D. Rossler, E. Soutschek, and R.C. Johnson, J. Clin. Microbiol. 33:103-109, 1995). Alignment with the OspC sequences of RFLP ribotype group IV, V, and VI isolates revealed that a sequence motif of all the isolates was quite similar to that of Borrelia garinii. A phylogenetic analysis based on OspC protein sequences also showed that most of the Japanese isolates were closely related to the species B. garinii. THe RFLP ribotype group IV species is predominant among clinical isolates of Lyme disease patients, reservoir rodents, and adult ticks in Japan. Although the isolates differed from type strains of the three delineated genospecies in genetic and immunological characteristics, it is likely that the spirochetes diverged within the species level. Therefore, the representatives of ribotype groups IV, V, and VI appear to have evolved within B. garinii and to have adapted to an Asiatic habitat, and there appeared to be a sufficient ecological pressure to allow bacterial species level development. PMID- 7494037 TI - Use of a specific immunogenic region on the Cowdria ruminantium MAP1 protein in a serological assay. AB - Currently available serological tests for cowdriosis (Cowdria ruminantium infection) in domestic ruminants are hampered by their low specificities because of cross-reactivity with Ehrlichia spp. The use of recombinant major antigenic protein (MAP1) of C. ruminantium for serodiagnosis was investigated. Overlapping fragments of the MAP1 protein were expressed in Escherichia coli and were reacted with sera from sheep infected with either C. ruminantium or Ehrlichia ovina. Two immunogenic regions on the MAP1 protein, designated MAP1-A and MAP1-B, were identified. MAP1-A was reactive with C. ruminantium antisera, E. ovina antisera, and three MAP1-specific monoclonal antibodies, whereas MAP1-B reacted only with C. ruminantium antisera. An indirect enzyme-linked immunosorbent assay (ELISA) based on MAP1-B was further developed and validated with sera from animals experimentally infected with C. ruminantium or several Ehrlichia spp. Antibodies raised in sheep, cattle, and goats against nine isolates of C. ruminantium reacted with MAP1-B. Cross-reactivity with MAP1-B was limited to Ehrlichia canis and Ehrlichia chaffeensis, two rickettsias which do not infect ruminants. Antibodies to Ehrlichia spp. which do infect ruminants (E. bovis, E. ovina, and E. phagocytophila) did not react with MAP1-B. Antibody titers to C. ruminantium in sera from experimentally infected cattle, goats, and sheep were detectable for 50 to 200 days postinfection. Further validation of the recombinant MAP1-B-based ELISA was done with sera obtained from sheep raised in heartwater-free areas in Zimbabwe and from several Caribbean islands. A total of 159 of 169 samples which were considered to be false positive by immunoblotting or indirect ELISA did not react with MAP1-B. In conclusion, recombinant MAP1-B may be a suitable antigen for a sensitive serological test for cowdriosis, with dramatically improved specificity. PMID- 7494040 TI - Detection of Toxoplasma gondii by PCR and tissue culture in cerebrospinal fluid and blood of human immunodeficiency virus-seropositive patients. AB - To investigate whether both tissue culture and PCR on a sequence from the repetitive rDNA could contribute to the diagnosis of toxoplasmosis, blood samples and, if they were available, cerebrospinal fluid (CSF) and aqueous humor samples from 72 human immunodeficiency virus-seropositive patients with suspected toxoplasmosis were prospectively tested. For 10 patients with fever of unknown origin but without confirmed toxoplasmosis, no Toxoplasma gondii was detected. For two patients with confirmed toxoplasmic uveitis, only PCR of aqueous humor samples was positive. Of 60 patients (48 with CSF samples) with neurological signs, 25 (from 13 of whom CSF samples were available) had confirmed cerebral toxoplasmosis and 10 had a positive PCR of CSF and/or blood samples, while for 1 patient culture of the CSF sample was also positive. Unlike tissue culture, PCR of rDNA is of value for the detection of cerebral toxoplasmosis in human immunodeficiency virus-seropositive patients, provided that both CSF and blood samples are available (sensitivity, 76.9%; specificity, 100%). PMID- 7494041 TI - Identification of novel insertion elements, restriction fragment length polymorphism patterns, and discontinuous 23S rRNA in Lyme disease spirochetes: phylogenetic analyses of rRNA genes and their intergenic spacers in Borrelia japonica sp. nov. and genomic group 21038 (Borrelia andersonii sp. nov.) isolates. AB - Borrelia spp. associated with Lyme disease possess an rRNA gene organization consisting of a single 16S rRNA gene followed by a spacer of several kilobases and a tandem repeat of a 23S (rrl)-5S (rrf) rRNA gene cluster. The restriction fragment length polymorphism (RFLP) patterns for these genes have been widely used to classify Lyme disease spirochete isolates. We analyzed the rRNA gene organization and sequences for two Ixodes ovatus isolates from Japan (IKA2 and HO14) and two group 21038 isolates associated with Ixodes dentatus ticks or rabbits from North America (isolates 21038 and 19857). This analysis revealed unique polymorphisms not previously described in other Lyme disease spirochete isolates. The molecular basis of these polymorphisms was determined by Southern blotting and PCR analyses. Only one continuous copy of the rrl-rrf gene cluster was identified in isolates IKA2, 19857, and 21038. The second rrl-rrf gene cluster is entirely absent from the IKA2 genome. In isolates 19857 and 21038, an intervening sequence is present, resulting in a fragment rrlB gene. The insertion site of this intervening sequence element differed in each isolate. While isolates 19857 and 21038 were found to carry a fragmented rrlB gene, they lacked rrfB. To determine if these rRNA polymorphisms were indicative of an underlying phylogenetic divergence, sequence analysis of the 16S rRNA (rrs) genes was conducted. The phylogenies inferred from rrs sequence analysis suggest that the polymorphisms resulted from recent mutational events. In addition, the phylogenetic analyses also support the proposed species status of Borrelia japonica sp. nov. and indicate that isolates of genomic group 21038 belong to a previously undescribed species for which we propose the nomenclature Borrelia andersonii sp. nov. PMID- 7494042 TI - Comparison of MICs of ceftiofur and other antimicrobial agents against bacterial pathogens of swine from the United States, Canada, and Denmark. AB - The MICs of ceftiofur and other antimicrobial agents, tested for comparison, for 515 bacterial isolates of pigs from the United States, Canada, and Denmark with various diseases were compared. The organisms tested included Actinobacillus pleuropneumoniae, Escherichia coli, Pasteurella multocida, Salmonella choleraesuis, Salmonella typhimurium, Streptococcus suis, Streptococcus dysgalactiae subsp. equisimilis, Streptococcus equi subsp. equi, and Streptococcus equi subsp. zooepidemicus. In addition to ceftiofur, the following antimicrobial agents or combinations were tested: enrofloxacin, ampicillin, sulfamethazine, trimethoprim-sulfadiazine (1:19), erythromycin, lincomycin, spectinomycin, lincomycin-spectinomycin (1:8), tilmicosin, and tetracycline. Tilmicosin was only tested against the U.S. isolates. Overall, ceftiofur and enrofloxacin were the most active antimicrobial agents tested against all isolates, with MICs inhibiting 90% of isolates tested (MIC90s) of < or = 2.0 and < or = 1.0 microgram/ml, respectively. Erythromycin, sulfamethazine, spectinomycin, and lincomycin demonstrated limited activity against all of the organisms tested, with MIC90s of > or = 8.0, > or = 256.0, > or = 32.0, and > or = 16.0 micrograms/ml, respectively. Trimethoprim-sulfadiazine was active against isolates of A. pleuropneumoniae, S. choleraesuis, S. typhimurium, P. multocida, S. equi, and S. suis (MIC90s, < or = 0.5 microgram/ml) but was less active against the E. coli strains tested (MIC90, > 16.0 micrograms/ml). Ampicillin was active against the P. multocida, S. suis, and S. equi isolates tested (MIC90s, 0.5, 0.06, and 0.06 micrograms/ml, respectively) and was moderately active against S. typhimurium (MIC90s, 2.0 micrograms/ml). However, this antimicrobial agent was much less active when it was tested against A. pleuropneumoniae, S. cholerae-suis, and E. coli (MIC90s, 16.0, > 32.0, and 32.0 micrograms/ml, respectively). Against the U.S. isolates of A. pleuropneumoniae and P. multocida, tilmicosin was moderately active (MIC90s, 4.0 and 8.0 micrograms/ml, respectively). However, this compound was not active against the remaining U.S. isolates (MIC90s, > 64.0 micrograms/ml). Differences in the MICs from one country to another were not detected with enrofloxacin, ceftiofur, or lincomycin for the strains tested, but variations in the MICs of the remaining antimicrobial agents were observed. PMID- 7494044 TI - Evaluation of BACTEC 9240 blood culture system by using high-volume aerobic resin media. AB - The BACTEC 9240 blood culture system (Becton Dickinson Diagnostic Instrument Systems, Sparks, Md.) is one of three automated, continuous-monitoring systems that is widely used in clinical laboratories. The BACTEC 9240 was compared with the BACTEC NR 660 for the detection of organisms and bacteremic episodes; time to detection of positive cultures; number of false-positive and false-negative cultures; and time needed to load, process, and perform quality control functions by using high-volume aerobic media. Blood specimens (5,282) were inoculated in equal volumes (5 to 10 ml per bottle) into BACTEC Plus Aerobic/F (9240 system) and BACTEC Plus NR26 (660 system) bottles. Clinically significant isolates were detected in 6.6% of cultures, representing 348 microorganisms and 216 bacteremic episodes. Two hundred forty-eight microorganisms were detected by both systems, 48 by the 9240 only and 52 by the 660 only (P = not significant). Of the bacteremic episodes, 158 were detected by both systems, 27 by the 9240 only and 31 by the 660 only (P = not significant). Analysis of data by month revealed equivalent recovery rates for both systems, with the exception of a 30-day period at one study site during which the 660 system detected significantly more microorganisms. Following a proprietary hardware design retrofit of the 9240 instrument, detection rates were again equivalent for the remaining three months at this study site. Positive cultures detected by both systems were detected an average of 4.3 h faster by the 9240 system (21 versus 25.3 h). The numbers of false-positive cultures for the 9240 and 660 systems were 40 (1.0%) and 9 ( < 1.0%), respectively. The numbers of false-negative cultures were five for the 9240 system and three for the 660 system. The 9240 system required 23 s less technologist time per bottle to operate during the 5-day protocol. In conclusion, the BACTEC 9240 used with high-volume Aerobic/F medium is equivalent to the BACTEC 660 used with high volume NR26 medium for the detection of microorganisms and bacteremic episodes. In addition, the 9240 system detects positive cultures more rapidly than the 660 system but requires further evaluation to ensure reliability of instrument components. PMID- 7494043 TI - Bartonella henselae prevalence in domestic cats in California: risk factors and association between bacteremia and antibody titers. AB - The isolation of Bartonella henselae, the agent of cat scratch disease, from the blood of naturally infected domestic cats and the demonstration that cats remain bacteremic for several months suggest that cats play a major role as a reservoir for this bacterium. A convenience sample of 205 cats from northern California was selected between 1992 and 1994 to evaluate the B. henselae antibody and bacteremia prevalences and to determine the risk factors and associations between bacteremia and antibody titers. B. henselae was isolated from the blood of 81 cats (39.5%). Forty-two (52%) of these bacteremic cats were found to be infected with > or = 1,000 CFU/ml of blood. Impounded or former stray cats were 2.86 (95% confidence interval [CI] = 1.94, 4.22) times more likely to be bacteremic than the pet cats. Young cats ( < 1 year old) were more likely than adult cats to be bacteremic (relative risk = 1.64; (95% CI = 1.19, 2.28). Bacteremic cats were more likely than nonbacteremic cats to be infested with fleas (relative risk = 1.64; 95% CI = 1.38, 1.96). No association between B. henselae infection and feline immunodeficiency virus antibody prevalence was observed. Eighty-one percent of the cats (166 of 205) tested positive for B. henselae antibodies, and titers were higher in bacteremic than in nonbacteremic cats. Multiple logistic regression analysis indicated that younger age and seropositivity for B. henselae antibodies were associated with bacteremia. Serological screening for Bartonella antibodies may not be useful for the identification of bacteremic cats (positive predictive value = 46.4%), but the lack of antibodies to B. henselae was highly predictive of the absence of bacteremia (negative predictive value = 89.7%). Seronegative cats may be more appropriate pets for immunocompromised individuals who are at increased risk for developing severe B. henselae disease. PMID- 7494045 TI - Evaluation of an enterovirus group-specific anti-VP1 monoclonal antibody, 5-D8/1, in comparison with neutralization and PCR for rapid identification of enteroviruses in cell culture. AB - We evaluated the usefulness of a commercially available monoclonal antibody (MAb) directed against a group-specific epitope of the capsid protein VP1 of enteroviruses for the rapid identification of these viruses in cell culture. The MAb was assayed in an indirect immunofluorescence test with cultured cells infected by various serotypes of enterovirus; all 39 serotypes tested, including echoviruses 22 and 23, which are considered atypical enteroviruses, were reactive. The MAb was also tested with 61 strains recovered from clinical specimens inoculated into cell cultures in comparison with seroneutralization with intersecting pools of hyperimmune sera and PCR with primers from the 5' untranslated region of enteroviruses. There was total agreement between the results obtained with the MAb and those obtained by PCR, even for those strains of enteroviruses which were found to be untypeable with polyclonal antisera. These data demonstrate the usefulness of the MAb for rapid identification of enteroviruses in cell culture. PMID- 7494046 TI - Phenotypic characteristics of 31 strains of Corynebacterium striatum isolated from clinical samples. AB - During a 34-month period (January 1991 to October 1993), 31 Corynebacterium striatum stains recovered from clinical samples from 24 patients were characterized. Twenty (64%) strains were isolated from wound exudates, 5 (16%) were isolated from bronchial aspirates, 2 (7%) were isolated from urine, 2 (7%) were isolated from endotracheal tubes, 1 (3%) was isolated from a catheter, and 1 (3%) was isolated from empyema. The organisms were identified by conventional culture and phenotypic characterization, the API CORYNE system, and cellular fatty acid composition analyses. The colonies of C. striatum could be confused with those of coagulase-negative staphylococci upon primary isolation from clinical material. A consistent phenotypic pattern was observed: all strains reduced nitrate, hydrolyzed tyrosine, and produced acid from glucose, fructose, and sucrose but not from maltose. API CORYNE profile numbers were 3100105 (28 strains) and 3000105 (3 strains). Susceptibility testing of C. striatum was performed by disk diffusion. All strains were susceptible to both imipenem and vancomycin and resistant to fosfomycin; most strains were susceptible to ampicillin and cephalosporins and resistant to clindamycin, erythromycin, and tetracycline. Performing a Gram stain of fosfomycin-resistant "Staphylococcus like" colonies was critical in order to identify C. striatum. PMID- 7494047 TI - Phenotypic and genotypic characterization of unusual vaginal isolates of Candida albicans from Africa. AB - As expected by its global prevalence, the most frequently isolated species of yeast from vaginal swabs obtained from patients in Africa was Candida albicans, which accounted for 53 of 85 (62.4%) of the isolates from women in Madagascar and 35 of 54 (64.8%) of the culture-positive women in Angola. However, 40% of the Madagascan and 23% of the isolates from Angola, as well as two isolates obtained from one German patient, were not able to utilize the amino sugars glucosamine and N-acetylglucosamine as the sole carbon source. These isolates were able to form germ tubes but did not form chlamydospores. The correct identification as C. albicans was made possible only by using a PCR-based method of DNA fingerprinting. Only minor phenotypic and genotypic variation was observed among these strains. Whether they represent a distinct clone that is found mainly in Africa is not clear. The relevance of the amino sugar catabolic pathway in C. albicans is discussed in view of these results. PMID- 7494048 TI - Development and evaluation of capture immunoglobulin G and M hemadherence assays by using human type O erythrocytes and recombinant parvovirus B19 antigen. AB - The capacity of human parvovirus B19 to agglutinate human type O erythrocytes was used to develop immunoglobulin G and M antibody capture hemadherence assays. When results of these assays were compared with those of corresponding antibody capture enzyme immunoassays using a well-characterized panel of 125 serum specimens, a 96.8% overall agreement was obtained between the two methods. PMID- 7494049 TI - Testing of susceptibility of Mycobacterium tuberculosis to pyrazinamide: comparison of Bactec method with pyrazinamidase assay. AB - The susceptibility of 428 clinical isolates of Mycobacterium tuberculosis to pyrazinamide was assessed by the Bactec method and the Wayne pyrazinamidase assay. The correlation between the two tests was 98.2 and 100% for susceptible and resistant strains, respectively. False resistance was seen in four (0.8%) strains with the Bactec test, and false-susceptible results occurred in two (0.5%) pyrazinamidase assays. The Bactec test is rapid and reliable, and the Bactec results correlate well with the pyrazinamidase test results, although some strains did not grow well in the test medium. PMID- 7494050 TI - Peritonitis involving a Capnocytophaga sp. in a patient undergoing continuous ambulatory peritoneal dialysis. AB - Capnocytophaga is a genus of gram-negative rods involved mainly in bacteremia in immunocompromised hosts. Here we report a case of Capnocytophaga peritonitis in a patient undergoing continuous ambulatory peritoneal dialysis. The disease appeared as a superinfection during antimicrobial therapy of a previous episode of peritonitis. PMID- 7494051 TI - A simple and sensitive DNA hybridization assay used for the routine diagnosis of human parvovirus B19 infection. AB - A dot blot hybridization assay for parvovirus B19 diagnosis was developed by using a PCR-generated probe, digoxigenin labelling, and chemiluminescence detection. Different labelling techniques and hybridization solutions were evaluated. From this analysis a protocol was devised for routine diagnostic use. The protocol enabled 1 pg of B19 DNA to be detected. The results of applying this method to 8,369 diagnostic samples collected during 1994 and 1995 are given. PMID- 7494053 TI - Relative value of selective group A streptococcal agar incubated under different atmospheres. AB - A commercially available selective group A streptococcal agar (ssA) was evaluated for the recovery of group A streptococci (GAS) in comparison with recovery from simultaneous cultures on conventional sheep blood agar (SBA). Both sets of plates were incubated in air, 5% CO2, and anaerobically for 48 h, with a first reading taken at 24 h. A total of 402 (67.0%) GAS were isolated from the 600 specimens that were submitted. Recovery of GAS was significantly greater after 48 h of incubation than after 24 h of incubation for each medium-atmosphere combination. After 48 h of incubation, the sensitivities of GAS detection obtained by each culture technique were as follows: ssA-anaerobic atmosphere, 98.5%; SBA-anaerobic atmosphere, 89.5%; ssA-CO2 atmosphere, 88.0%; SBA-air, 86.5%; SBA-CO2 atmosphere, 82.0%; and ssA-air, 74.6%. There were no cultures positive in air or CO2 which were not positive anaerobically on either medium. The increased sensitivity of detecting positive GAS cultures when incubation was done in an ssA-anaerobic atmosphere for 48 h uncovered patients truly infected with the organisms. PMID- 7494052 TI - Identification of Candida species by PCR and restriction fragment length polymorphism analysis of intergenic spacer regions of ribosomal DNA. AB - The PCR was used to amplify a targeted region of the ribosomal DNA from 84 Candida isolates. Unique product sizes were obtained for Candida guilliermondii, Candida (Torulopsis) glabrata, and Candida pseudotropicalis. Isolates of Candida albicans, Candida tropicalis, Candida stellatoidea, Candida parapsilosis, and Candida krusei could be identified following restriction digestion of the PCR products. PMID- 7494054 TI - Screening urine samples by leukocyte esterase test and ligase chain reaction for chlamydial infections among asymptomatic men. AB - Urine samples from 358 asymptomatic males were screened for urethral inflammation by the leukocyte esterase (LE) test and for Chlamydia trachomatis by the ligase chain reaction (LCR). LE and LCR positivity rates were 7.5% (27 of 358 samples) and 2.8% (10 of 358 samples), respectively. Eight of the 10 LCR-positive samples were detected by the LE screening test. The urine LE prescreening test in combination with the LCR assay may be a reasonable approach for genitourinary chlamydial disease control. PMID- 7494055 TI - Detection of measles virus RNA in urine specimens from vaccine recipients. AB - Analysis of urine specimens by using reverse transcriptase-PCR was evaluated as a rapid assay to identify individuals infected with measles virus. For the study, daily urine samples were obtained from either 15-month-old children or young adults following measles immunization. Overall, measles virus RNA was detected in 10 of 12 children during the 2-week sampling period. In some cases, measles virus RNA was detected as early as 1 day or as late as 14 days after vaccination. Measles virus RNA was also detected in the urine samples from all four of the young adults between 1 and 13 days after vaccination. This assay will enable continued studies of the shedding and transmission of measles virus and, it is hoped, will provide a rapid means to identify measles infection, especially in mild or asymptomatic cases. PMID- 7494056 TI - Prior enrichment of human immunodeficiency virus DNA with probe DNA particles for efficient PCR diagnosis. AB - We report here a novel method by which one can recover human immunodeficiency virus (HIV) DNA in a small volume (approximately 50 microliters) of solution from 8 ml of crude cell lysate which contains as few as several HIV DNA copies. The method uses the specific binding of HIV DNA to HIV probe DNA particles. The HIV DNA thus concentrated on the particles can be subjected to the PCR assay. The method also enables the examination of 100 individual blood specimens in a combined form. PMID- 7494057 TI - Streptococcus pneumoniae types 19A and 19F and Streptococcus suis capsular type 8 share common capsular epitopes. AB - Two monoclonal antibodies (MAbs) to Streptococcus pneumoniae types 19A and 19F were tested with the 35 reference strains and 334 field strains of Streptococcus suis by dot blotting. Both MAbs reacted with the capsular type 8 reference strain, and one reacted with 69% and one reacted with 100% of 81 S. suis capsular type 8 field strains tested. Epitopes recognized by both MAbs are capsular in origin. PMID- 7494058 TI - Characterization of chromosomal DNA profiles from Helicobacter pylori strains isolated from sequential gastric biopsy specimens. AB - The restriction endonuclease profiles of DNAs from Helicobacter pylori strains isolated from 20 patients in two or more consecutive biopsy specimens over a period of up to 2 years were analyzed by pulsed-field gel electrophoresis with NotI and NruI. H. pylori strains possess a high degree of genomic diversity which was not observed to occur in vivo, and attempts to observe it in vitro were not successful. PMID- 7494059 TI - Immunological detection of penicillin-binding protein 2' of methicillin-resistant staphylococci by using monoclonal antibodies prepared from synthetic peptides. AB - Two synthetic peptides 31 and 32 amino acids in length were prepared as deduced from a known amino acid sequence of penicillin-binding protein 2' (PBP2') of methicillin-resistant Staphylococcus aureus. Two monoclonal antibodies were generated from fused cells of myeloma cells and splenic cells of mice immunized with the synthetic peptides. Western blot (immunoblot) analysis demonstrated specific binding of the antibodies to PBP2' of a methicillin-resistant S. aureus strain. An immunoradiometric assay was developed by using these antibodies for simple detection of PBP2'. PMID- 7494060 TI - Detection of Cowdria ruminantium in blood and bone marrow samples from clinically normal, free-ranging Zimbabwean wild ungulates. AB - Cowdria ruminantium causes severe, often fatal disease in domestic ruminants, whereas wildlife species usually are not affected. Blood and bone marrow samples from healthy, free-ranging Zimbabwean ungulates were taken during translocation from areas harboring Amblyomma ticks and tested for the presence of C. ruminantium, using a PCR assay based on the C. ruminantium map1 gene. Positive reactions were obtained in tsessebe (Damaliscus lunatus), waterbuck (Kobus ellipsiprymnus), and impala (Aepyceros melampus). Wildlife species may therefore be a reservoir for C. ruminantium thus contributing to the spread of cowdriosis. PMID- 7494061 TI - Survival of anaerobic bacteria in various thioglycolate and chopped meat broth formulations. AB - Three commercially available formulations of thioglycolate broth and of chopped meat broth were evaluated for their abilities to maintain the viabilities of 32 strains of anaerobic bacteria during a period of 8 weeks. While thioglycolate broth supported the initial (48-h) growth of all strains tested, approximately half of the strains died off within 4 weeks. Chopped meat broths maintained the viabilities of almost all cultures during the test period. PMID- 7494062 TI - Catheter-related bacteremia associated with coagulase-positive Staphylococcus intermedius. AB - We report a case of catheter-related bacteremia in a 63-year-old patient caused by Staphylococcus intermedius. Clinical resolution of the infection was obtained after removal of the intravenous device and antibiotic treatment. This observation emphasizes the risk of confusion between S. intermedius and Staphylococcus aureus if only a coagulase test is done. PMID- 7494063 TI - Monoclonal antibodies for detection of Norwalk virus antigen in stools. AB - Monoclonal antibodies against the prototype 8FIIa strain of Norwalk virus were prepared and applied to an enzyme immunoassay (EIA) for detecting Norwalk virus in stool specimens. The monoclonal antibodies immunoprecipitated a 58-kDa protein which had been produced by in vitro transcription-translation of Norwalk virus cloned cDNA, and they reacted by EIA with recombinant Norwalk virus capsid protein at a sensitivity level of 1 ng/ml. The EIA detected virus in all tested samples from 15 different Norwalk virus-infected volunteers. No cross-reactions were seen in stools containing other caliciviruses or in stools containing rotaviruses, astroviruses, or enteric adenoviruses. PMID- 7494064 TI - Species-specific Mycobacterium genavense DNA in intestinal tissues of individuals not infected with human immunodeficiency virus. AB - Mycobacterium genavense species-specific DNA was detected in intestinal tissues from two of nine individuals not infected with human immunodeficiency virus. This newly described microorganism is well documented as a causative agent of disseminated infections in AIDS patients. Our results suggest that it may colonize the guts of individuals not infected with human immunodeficiency virus. PMID- 7494065 TI - Comparison of recovery rates for mycobacteria from BACTEC 12B vials, Middlebrook 7H11-selective 7H11 biplates, and Lowenstein Jensen slants in a public health mycobacteriology laboratory. AB - Recovery rates from Middlebrook 7H11-selective 7H11 biplates and Lowenstein Jensen slants (LJ) used with BACTEC 12B vials were compared for 5,399 specimens. For 578 specimens that were inoculated onto three media, 580 mycobacteria were isolated, including 277 (48%) Mycobacterium avium complex isolates, 230 (40%) Mycobacterium tuberculosis isolates, and 73 (12%) other mycobacteria. For BACTEC 12B vials, 506 (87%) cultures were positive, 45 (8%) were negative, and 29 (5%) were lost to contamination; for 7H11-7H11-selective biplates, 469 (81%) cultures were positive, 95 (16%) were negative, and 16 (3%) were lost to contamination; for LJ, 230 (40%) cultures were positive, 111 (19%) were negative, and 239 (41%) were lost to contamination. For routine cultures, use of plate media is superior to use of LJ. PMID- 7494066 TI - Production and characterization of monoclonal antibodies against Serpulina hyodysenteriae and S. innocens and their use in serotyping. AB - Murine monoclonal antibodies (MAbs) directed against serotypes 1, 2, 8, and 9 of Serpulina hyodysenteriae and strain B256 of Serpulina innocens were produced and characterized. A serological classification of 96 field strains of S. hyodysenteriae and 28 field strains of S. innocens isolated from pigs showing clinical signs of swine dysentery was performed by rapid dot enzyme-linked immunosorbent assay (ELISA) with the MAbs. The results indicated that the majority of the field strains of S. hyodysenteriae (69%) belonged to serotypes 8, 1, and 9, whereas only 31% of the S. innocens strains were recognized by MAb 9H7, indicating the presence of antigenic heterogeneity among S. innocens isolates. Rapid dot ELISA with type-specific MAbs was found to be specific, sensitive, and easy to perform and thus to be suitable for routine serotyping of S. hyodysenteriae and S. innocens isolates. This is the first report of MAbs being used for serotyping clinical isolates of S. hyodysenteriae and S. innocens. PMID- 7494067 TI - Detection of hepatitis C virus RNA by methods using phenol-chloroform or silica particles. PMID- 7494069 TI - Pilot testing an independent study series on research. AB - The Army Nurse Corps developed The Critical Reading of Research Publications (CRRP) program, an independent study series for small groups. The goal of the program is to improve patient care by providing a means for nurses to learn basic research principles in a self-paced format, while reading and discussing nursing research articles of clinical relevance. Continuing education (CE) approval of the program required pilot testing to evaluate its ease of use, effectiveness, and allocation of CE contact hours. The purpose of pilot testing was to improve both the participants' experience and the administrative management of the CE program. The constructive feedback received by potential consumers resulted in incremental improvement of an innovative educational program in nursing research. PMID- 7494068 TI - Competence: an evolving concept. AB - Formal and informal definitions of competence are the basis for building a useful understanding of the concept. However, further dissection and analysis are necessary before the concept of competence can be rebuilt into a new and more meaningful whole useful in theory development and hypothesis generation around staff development and continuing education in nursing. Reflection, analysis, thought, perception, visualization, and contextualization of the concept of competence are the foci of this article. PMID- 7494070 TI - Is research in the practice setting feasible? AB - Staff development educators are in key positions to assist staff nurses with practice-oriented research, a necessary part of clinical practice. The first step in developing an environment conducive to practice-oriented research is the establishment of a "working committee" within existing quality assurance or standards committees. As committee chairpersons, staff development educators can help staff nurses recognize the research potential inherent in practice by relating research terminology to the nursing process. Examples of client care problems illustrate how nurses can exert control over their practice and effect positive change. PMID- 7494071 TI - Documenting patient education: a literature review. AB - Lack of time is cited as the most common reason why staff nurses neglect to document patient education. Consumer rights, standards of practice, and other laws regulating nursing practice require nurses to teach patients. Administration and staff development can enhance nurses' efforts to document more effectively. Forms and flowsheets detailing content and learner outcomes and tracking patients' progress can affect the quality of documentation. Nurses are encouraged to continue developing new forms and flowsheets and to share their products with others. PMID- 7494072 TI - Development of a nursing care manual to improve the knowledge of nurses caring for hospitalized patients with diabetes. AB - Nurses have a responsibility to ensure that they have adequate knowledge in order to provide competent care. A wide literature search failed to identify adequate documentation of the specific nursing care of people with diabetes, although the medical aspects are covered in detail. In addition, several papers document a deficiency of knowledge among nurses about diabetes and the appropriate care of patients with diabetes. This project sought to address the apparent lack of documentation. The aim was to formulate a Manual of Nursing Care of People with Diabetes Mellitus (the manual), to be used as a self-directed learning resource in the clinical setting. This article reports the development and evaluation of the manual. PMID- 7494073 TI - The time has come--a study of geriatric continuing education in Ohio Visiting Nurse Associations. AB - A majority of nurses working in the community lack formal educational preparation to care for older adults. This study examined the perceived learning needs of Visiting Nurse Association (VNA) executives and agency staff nurses in Ohio related to continuing education in gerontology. Twenty-six executives and 478 staff nurses were asked to complete a gerontologic education survey; 17 executives and 344 staff nurses returned surveys. Findings suggest that few Ohio VNAs have focused on continuing education in gerontology. Indications exist that executives and staff nurses are not always in agreement regarding the importance of content needed for updating gerontological knowledge and skills. Educational and organizational strategies are offered to provide minimal standards for gerontologic professional practice in home care. PMID- 7494074 TI - Developing a teaching video on mechanical ventilation weaning to facilitate effective nursing research. AB - This article describes the process of developing a teaching video associated with conducting a nursing research study and directed at educating critical care staff nurses about the process of successfully weaning patients from mechanical ventilation. Techniques on developing the script and filming and editing the video are presented. The results of this process included staff empowerment and a teaching video used by 10 hospitals in a multi-site research project. PMID- 7494075 TI - Development and implementation of a leadership skills course for the charge nurse. AB - Formal education in leadership behaviors and skills is paramount to the development of staff nurses as effective leaders. The need for effective leadership during the present transformation of healthcare, and the Joint Commission for Accreditation of Health Care Organization's mandate to educate and train staff as appropriate to their job responsibilities, resulted in the development and implementation of a leadership skills course for the staff charge nurse. A literature review, input from experienced charge nurses, and clinical nurse managers' perspectives of what is effective leadership for a charge nurse played key roles in development and implementation of the course. PMID- 7494076 TI - Storytelling: a method to increase discussion, facilitate rapport with residents and share knowledge among long-term care staff. PMID- 7494077 TI - Ten steps to improving CE: a summary for learners. PMID- 7494078 TI - Practice parameters for the diagnosis and treatment of asthma. Joint Task Force on Practice Parameters, representing the American Academy of Allergy Asthma and Immunology, the American College of Allergy, Asthma and Immunology, and the Joint Council of Allergy, Asthma and Immunology. PMID- 7494079 TI - The development and significance of optometric clinical practice guidelines. PMID- 7494080 TI - A comparison of TPA/DPA certification and access to care in North Carolina, Missouri, and Illinois. AB - BACKGROUND: The changing scope of optometric practice has had a large impact on the profession. Optometry has gone from a drugless profession to a profession that can use diagnostic pharmaceutical agents (DPAs) in every state and therapeutic pharmaceutical agents (TPAs) in 46 states. The intent of this study was to determine the influence that these legal variables exert on the scope of practice for optometrists in different states. METHODS: Optometrists in three states with varying statutes regulating the scope of practice were surveyed to determine if there were practice differences related to these state laws. RESULTS: In states with TPA usage, more optometrists accepted Medicare assignment and had hospital and nursing home privileges. Optometrists with TPA certification had, as a group, graduated more recently. Travel times for patients with conditions requiring treatment with therapeutic drugs were shorter in the states with TPA legislation. CONCLUSIONS: Access to care is enhanced for citizens of a state with optometric usage of therapeutic pharmaceutical agents. PMID- 7494081 TI - Automaticity in fusional vergence therapy. AB - BACKGROUND: Fusional vergence therapy is typically performed under conditions that permit the patient to direct considerable attention and effort to the visual task. Unless visual function is sufficiently automatic as to require minimal attention and effort, attention directed toward visual function may detract from that available for cognitive processing. This report describes a method to develop greater automaticity of fusional vengence through the use of concurrent demands. METHODS: A retrospective pilot study of patient records was performed to assess the use of a concurrent motor demand (bouncing on a rebounder) to develop automaticity of fusional vergence. RESULTS: In most cases, patients who achieved adequate fusional vergence without the concurrent motor demand demonstrated a decrease in fusional vergence ranges when the motor demand was added, and then improved with practice so that vergence ranges while bouncing on the rebounder approached those achieved while seated. This pattern was obtained in 75 percent of the base-out and 57.1 percent of the base-in protocols. Some patients who showed a decrement in performance with introduction of the concurrent demand failed to improve with practice; this pattern was observed in 28.6 percent of the base-in protocols, but in none of the base-out protocols. Some patients showed no decrease in performance when the concurrent distracting task was introduced (observed in 25 percent of the base-out protocols and 14.3 percent of the base-in protocols). CONCLUSIONS: In most cases, maximum automaticity is not achieved when fusional vergence is trained under conditions that permit the patient to devote full attention to the vergence task. Introducing concurrent distractors may be desirable to automate performance and enable the patient to maintain fusional vergence even while attention is directed to the concurrent task. Heightened automaticity may reduce the effort and attention that must be directed toward binocular function, and hence maximize attentional capacity available for information-processing, comprehension, and performance on the task at hand. PMID- 7494082 TI - Is there a typical VDT patient? A demographic analysis. AB - BACKGROUND: The use of video display terminals (VDT) in the workplace is growing rapidly. Visual problems associated with computer use are increasing apace. Recognizing and addressing these needs are vital tasks for the optometric practitioner. This paper examines the demographics, prevalence of symptoms, and other work- and vision-related factors among VDT workers. Understanding the characteristics of a typical patient can aid in developing effective treatment protocols. METHODS: Twenty-two optometrists collected data from 324 patients who work on VDT screens for a minimum of 2 hours per day. A questionnaire was used to collect the data. Along with reporting the patients' dry manifest refraction results, the survey instrument also included a self-report of experienced symptoms. RESULTS: The prototypical patient, based on the total subject population, is a 38-year old, mildly myopic female who uses the computer screen about 5 hours per day. Average total VDT work experience is about 5 years. Most of the patients use full-color screens and sit between 21 and 22 inches away from the screen. A variety of visual symptoms is reported, with eyestrain (65 percent) and headaches (42 percent) being the most common complaints. CONCLUSIONS: Between 75 percent (results of this study) and 88 percent (reported by NIOSH) of patients who use a VDT regularly are likely to be symptomatic. This study has helped to identify the "typical" VDT user and the symptoms this user is most likely to report. The next step is to develop a diagnostic and treatment protocol by which optometrists can prescribe for and potentially prevent these computer-related vision problems. PMID- 7494083 TI - A retrospective study of eye disease among first grade children in Los Angeles. AB - BACKGROUND: The prevalence of ocular disease among children in one school district in Los Angeles, California was studied to better understand the types of eye disorders among this population as well as to develop appropriate preventive programs. METHODS: A computer-assisted retrospective analysis of date was performed from charts of 2,204 first grade children examined in the UCLA Mobile Eye Clinic. RESULTS: One or more ocular disorders were observed in 22.3 percent of the subjects. Uncorrected best monocular visual acuity was 20/40 or worse in 3.4 percent of the children. Refractive errors were diagnosed in 15.7 percent of the subjects, astigmatism in 7.6 percent, hyperopia in 6.2 percent, and myopia in 6.0 percent. Color vision deficiencies (red-green) were found in 2.6 percent of boys. The prevalences of heterophorias and heterotropias were 1.2 percent and 1.3 percent, respectively. CONCLUSIONS: The variety of ocular disorders diagnosed in this demographic setting attests to the importance of performing early and effective screening eye examinations for children. PMID- 7494084 TI - Vision impairment and corrective considerations of civil airmen. AB - BACKGROUND: Civil aviation is a major commercial and technological industry in the United States. The Federal Aviation Administration (FAA) is responsible for the regulation and promotion of aviation safety in the National Airspace System. To guide FAA policy changes and educational programs for aviation personnel about vision impairment and the use of corrective ophthalmic devices, the demographics of the civil airman population were reviewed. METHODS: Demographic data from 1971 1991 were extracted from FAA publications and databases. RESULTS: Approximately 48 percent of the civil airman population is equal to or older than 40 years of age (average age = 39.8 years). Many of these aviators are becoming presbyopic and will need corrective devices for near and intermediate vision. In fact, there has been approximately a 12 percent increase in the number of aviators with near vision restrictions during the past decade. Ophthalmic considerations for prescribing and dispensing eyewear for civil aviators are discussed. CONCLUSIONS: The correction of near and intermediate vision conditions for older pilots will be a major challenge for eye care practitioners in the next decade. Knowledge of the unique vision and environmental requirements of the civilian airman can assist clinicians in suggesting alternative vision corrective devices better suited for a particular aviation activity. PMID- 7494085 TI - Thalasselis syndrome and genetic theories on keratoconus. AB - BACKGROUND: Previous theories of the pathogenesis of keratoconus have only described partial aspects of the disease. METHODS: Studies are reviewed that have demonstrated a statistical and physiological relation between keratoconus, magnesium deficiency, type A behavior, and allergy, which constitute the Thalasselis Syndrome. RESULTS: The Thalasselis syndrome integrates old and new theories on keratoconus, mitral valve prolapse, and tetany/menopause. Keratoconus is also associated with genetic diseases like Down syndrome, Marfan syndrome, and other conditions related to collagen disturbance and biochemical abnormalities in protein synthesis. CONCLUSIONS: This syndrome suggests that the genetic disturbance that causes these alterations could be influenced by metabolic factors in which magnesium deficiency could be involved. PMID- 7494086 TI - Clinicolegal aspects of practice guidelines for pediatric eye and vision examination. AB - BACKGROUND: The American Optometric Association (AOA) recently joined a growing trend in the health care professions by adopting clinical practice guidelines. The goals of practice guidelines are to improve quality of care and to provide a more objective measure of standard of care for the legal system and the health insurance industry. Practice guidelines may eventually reduce the number and cost of malpractice actions. Historically, the standard of care for pediatric eye and vision examination has been the subject of controversy. Issues of disagreement include the amount of ocular health testing that should be performed and the determination of appropriate emphasis on functional vision. This paper summarizes the precedents that have established a legal standard of care for pediatric eye and vision examination. METHODS: A review of malpractice claims relevant to pediatric optometry was conducted. Public health policy, legal, and malpractice literature was also consulted for current views on practice guidelines. The recent AOA Optometric Clinical Practice Guidelines for Pediatric Eye and Vision Examination were compared to this review. RESULTS: Standards of care for pediatric eye examination should minimally include assessment of visual acuity, refractive error, binocular vision, functional vision, and eye health assessment including dilated funds examination with a binocular indirect ophthalmoscope. These standards are commensurate with the AOA Guidelines. CONCLUSIONS: The AOA Optometric Clinical Practice Guidelines for Pediatric Eye and Vision Examination is compared to the current standard of care for pediatric optometry. It is concluded that the Guidelines are appropriate. Optometrists should integrate these practices into patient care, and the optometric profession should seek to obtain wide spread acceptance of the Guidelines. PMID- 7494087 TI - Lid lesions suspected of malignancy. AB - BACKGROUND: This review describes four examples of lid lesions and their clinical or surgical management. METHODS: Close examination and careful observation of the lids and surrounding tissue is necessary for early detection and diagnosis of skin lesions. Although skin carcinomas around the lids are usually not life threatening, delayed diagnosis can lead to extensive surgical intervention and cosmetic reconstruction. This review describes the epidemiology and clinicopathologic basis for the development of lid carcinomas. RESULTS: The differential diagnosis of lid lesions is reviewed and current accepted methods of therapy are discussed. CONCLUSIONS: Lid lesions are typically classified into malignant and benign categories. Beyond this general classification, knowledge of initial appearance, progression and potential pathologic change is necessary. Clinical presentations and management of both malignant and benign lid lesions are discussed. PMID- 7494088 TI - Evaluation of non-radioactive labelling and detection of deoxyribonucleic acids. Part Two: Colorigenic methods and comparison with chemiluminescent methods. AB - The diagnosis of genetic infections and cancerous diseases is carried out more and more often at a molecular level using Southern's technique which is based on the use of 32P-labelled DNA. In order to circumvent the risks and rapid decrease in radioactivity associated with these latter techniques, new colorigenic methods have been developed. In this work, we describe the use of dTTP analogues (digoxigenin-dUTP and biotin-dUTP) for the labelling of probes and detection of target DNA. Using digoxigenin-11-dUTP, 0.1 aM of a 561 bp target DNA was detected by using a modified Southern procedure. The reliability and the high sensitivity of such methods make them a good tool for DNA investigation in research as well as in testing laboratories. PMID- 7494089 TI - Osmotic pressure method to measure salt induced folding/unfolding of bovine serum albumin. AB - A new approach has been developed to monitor protein folding by utilizing osmotic pressure and a range of salt concentrations in a well characterized protein, bovine serum albumin (BSA). It is hypothesized that both the 'effective' osmotic molecular weight, Ae, and the solute/solvent interaction parameter, I, in the empirical relation Msolvent/Msolute = (RT rho/Ae)1/pi + I [1] can be used as measures of protein folding. I is a measure of solvent perturbed by the solute and is thought to depend directly upon the solvent accessible surface area (ASA). It is reasoned that larger solvent accessible surface area of an unfolded or denatured protein should perturb more water and produce larger I-values. Thus I values allow calculation of a unfolded protein fraction, fua, due to changes in relative solvent accessible surface area. It has been observed that Ac decreases for filamentous, denatured proteins due to segmental motion of the molecule [2]. This allows calculation of unfolded protein fraction from the effective molecular weight, fum. Colloid osmotic pressure of BSA was measured in a range of salt concentrations at 25 degrees C, and pH = 7 (above the isoelectric point of BSA at pH = 5.4). Both S and I were used to monitor protein folding as the salt concentration was varied. In general, larger and variable I-values and smaller Ae were observed at salt concentrations less than 50 mmolal NaCl (Imax = 8.9), while constant I = 4.1 and Ae = 66,500 were observed above 50 mmolal NaCl. The two expressions for fractional unfolding (fua and fum) are in general agreement. Small differences in the parameters below 50 mmolal salt concentration are explained with well known shifts in the relative amounts of alpha-helix, beta sheet and random coil in denatured BSA. The relative amounts of these shifts agree with predictions in the literature attributed to continuous BSA expansion rather than an 'all-or-none' conversion. PMID- 7494090 TI - A rapid and convenient filter-binding assay for ras p21 processing enzyme farnesyltransferase. AB - Because it is the target for the development of anti-cancer agents, the mammalian cytosolic enzyme farnesyltransferase (FTase) has received significant attention in recent years. FTase catalyzes the transfer of a farnesyl group from farnesylpyrophosphate (FPP) to cysteine 185/186 at the carboxyl terminal end of ras proteins (ras p21), a reaction essential for the localization of ras p21 to the plasma membrane for their cellular functions including cell transformation in case of oncogenic ras p21. Here, we report the development of a rapid and convenient assay procedure for FTase using phosphocellulose paper which has a binding affinity for proteins. The FTase is assayed as the transfer of [3H]farnesyl group from [3H]FPP to the ras p21 at pH 7.4 and 37 degrees C in the presence of rat brain cytosol followed by the binding of radioactive farnesylated ras p21 to the phosphocellulose paper. The radioactivity associated with ras p21 bound to the phosphocellulose paper was determined by scintillation counting after soaking the paper in trichloroacetic acid and washing with distilled water. Utilizing [3H]FPP and recombinant Ha-ras p21 as substrates in the reaction, the FTase followed Michaelis-Menten kinetics with Km values of 1.0 and 7.69 microM for respectively [3H]FPP and recombinant Ha-ras p21. The method reported here has the advantages over the other published assay procedures of being rapid, convenient and economical, and can be successfully used for the basic assaying of FTase in different organs and distinct species and for the screening of novel inhibitors of FTase. PMID- 7494091 TI - High-performance liquid chromatography of amino acids after derivatization with 9 isothiocyanatoacridine. AB - This study describes a reproducible method for the routine determination of amino acids in protein hydrolysates using 9-isothiocyanatoacridine as an HPLC precolumn derivatization reagent. Derivatization is carried out by adding the isothiocyanate dissolved in dry acetonitrile to a buffered amino acid solution without the following extraction of the excess reagent. The resulting N-(9 acridinylthiocarbamoyl)amino acids were separated by one single isocratic HPLC analysis with the UV detection at 280 nm. PMID- 7494092 TI - A simple and inexpensive sample-handling method for the semi-preparative RP-HPLC of polypeptides and non-polar peptide derivatives: pre-adsorption of samples. AB - A simple method is described for the application onto HPLC columns of very crude or alternatively poorly soluble polypeptide samples prior to their chromatographic purification. The procedure involves the batch pre-adsorption of the crude polypeptide mixture from a dilute solution onto an appropriate preparative-grade chromatographic adsorbent, removal of the solvent by rotary evaporation or lyophilisation and then dry-packing the pre-adsorbed chromatographic material into guard column cartridges of suitable dimensions. The polypeptide products can then be eluted either by isocratic or gradient elution methods through the cartridge coupled in tandem with prepacked semi-preparative HPLC columns. This method has been successfully utilised for the routine RP-HPLC purification of polar and hydrophobic polypeptides prepared by solid phase peptide synthesis (SPPS) methods as well as peptide derivatives and intermediates used as part of SPPS procedures. PMID- 7494093 TI - Base-pair exchange kinetics of the imino and amino protons of the 3'-phenazinium tethered DNA-RNA duplex, r(5'GAUUGAA3'):d(5'TCAATC3'-Pzn), and their comparison with those of B-DNA duplex. AB - The dynamics of the opening-closing of the constituent base-pairs as well as of the exchange kinetics of the base-paired imino and amino protons with water in a DNA-RNA hybrid, [5'r(G1A2U3U4G5A6A7)3']:5'p[d(T8C9A10A11T12C13)]3'-Pzn ] duplex (I), are reported here in details for the first time. The exchange kinetics of amino and imino protons in the DNA-RNA hybrid (duplex I) have been compared with identical studies on the following B-DNA duplexes: d(C1G2T3A4C5G6)2 (II), d[p(5'T1G2T3T4T5G6G7C8)3']:d[p(5'C9C10A11A12A13C14A15)3'] (III), d(C5G6C7G8A9A10T11T12C13G14C15G16)2 (IV) and d(C1G2C3G4C5G6C7G8A9A10T11T12C13G14C15G16C17G18C19G20)2 (V). This comparative study shows that the life-times tau o of various base-pairs in the DNA-RNA hybrid (I) varies in the range of approximately 1 ms, and they are quite comparable to those of the shorter B-DNA duplexes (II) and (III), but very different from the tau o of the larger duplexes (IV) and (V): the tau o for the base pair of T11 and T12 residues in the 20-mer (duplex V) are 2.9 +/- 2.3 ms and 23.2 +/- 8.9 ms, respectively, while the corresponding tau o in the 12-mer (duplex IV) are 2.8 +/- 2.2 ms and 17.4 +/- 5.4 ms. It has also been shown that the total energy of activation (Ea) assessed from the exchange rates of both imino and amino protons, representing energetic contributions from both base-pair and helix opening closing as well as from the exchange process of the imino protons from the open state with the bound water, is close to the Ea of the short B-DNA duplex (Ea approximately 28-47 kcal/mol). PMID- 7494094 TI - Enzymatic determination of carbon (14C)-labeled glycerol in biological samples. AB - A method for determination of glycerol-specific-radioactivity in biological samples is presented. It is based on the following steps: (a) enzymatic conversion of glycerol to dihydroxyacetone-phosphate, (b) quantitative trapping of dihydroxyacetone-phosphate in SPE amino (NH2) columns, (c) eluation with HCl 0.5 N of dihydroxyacetone-phosphate followed by radioactivity counting and (d) estimation of the radioactivity thus trapped compared with that of enzymatically untreated aliquots of the same samples. No interferences from other 14C-labeled materials tested such as D-glucose, L-alanine, L-glutamine and D-beta hydroxybutyrate were observed. This inexpensive and high-speed method can be applied in routine multiple estimations of glycerol-specific-radioactivity in biological samples in tracer metabolic studies. PMID- 7494095 TI - A method for in vivo [32P]phosphate labelling of testis proteins. AB - The direct intratesticular injection of [32P]phosphate resulted in 4-9 times more labelling of rat testis proteins compared to the conventional method of in vitro incubation. Moreover this is a simple technique requiring minimum (7-10 times less) radioactive phosphate and is less hazardous. PMID- 7494096 TI - Labeled adenosine(5')tetraphospho(5')adenosine (Ap4A) and adenosine(5')tetraphospho(5')nucleoside (Ap4N). Synthesis with firefly luciferase. AB - Labeled dinucleoside polyphosphates are not commercially available, in spite of being important molecules in metabolic regulation. Firefly luciferase (EC 1.13.12.7) is a useful enzyme for the synthesis of adenosine(5')tetraphospho(5')adenosine (Ap4A). As luciferase behaves as a nucleotidase at low ATP concentration, adequate concentrations (higher than 0.1 mM ATP) should be used to obtain a good yield of labeled Ap4A. [32P]Ap4A has also been synthesized from ATP and [32P]PPi. In a first step, [beta, gamma-32P]ATP is generated in a ATP-[32P]PPi exchange reaction catalyzed by luciferase. In a second step, the reaction is supplemented with pyrophosphatase and 32P labeled Ap4A is obtained. Radioactive adenosine(5')tetraphospho(5')nucleoside (Ap4N) can also be synthesized from ATP gamma S and labeled NTP or from low concentrations of labeled ATP and high concentrations of cold NTP. The syntheses of radioactive ApnA and pnA (n > 4) can also be approached with luciferase. PMID- 7494097 TI - Quantification of protein in dilute and complex samples: modification of the bicinchoninic acid assay. AB - The colorimetric assay using bicinchoninic acid (BCA) as test reagent is useful for quantitative protein determinations due to its high sensitivity, ease, and tolerance to various contaminations present in biological samples or added during purification. For removal of interfering substances, protein precipitations have been described. Yet, obstructions became apparent with diluted and complex samples. Therefore we tested different solvents for removal of such interfering contaminants from the protein precipitate, and 1 M HCl was identified as the useful washing agent. The protocol described allows simple and accurate microdetermination in microtiter plates of proteins from complex samples. PMID- 7494098 TI - Determination of triasulfuron in soil: affinity chromatography as a soil extract cleanup procedure. AB - Triasulfuron forms one of the group of sulfonylurea herbicides. These are used widely for controlling weeds as they are effective at very low application rates. This effectiveness is responsible for the crop losses due to persistence of trace amounts of the herbicides (< or = 100 pg/g) in the soil. The numerous immunoassays described have been constrained by the fact that the soil extract contains co extractants which interfere in the assays, so much so, that these assays are useless at low levels of herbicide. We describe here the preparation and application of an immunoaffinity column which binds specifically to triasulfuron, thus cleaning up the soil extract. The experiment design is such that this also leads to concentration of the triasulfuron, making it easier to assay reliably using ELISA. Six different soil types were used to validate this procedure. In most cases, the herbicide content could be detected at 100 pg/g (critical phytotoxic herbicide level in soil) with a variation of +/- 20% in the readings. PMID- 7494099 TI - Evaluation of non-radioactive labelling and detection of deoxyribonucleic acids. Part One: Chemiluminescent methods. AB - The growth of analytical methods for the detection of nucleic acid from various biological samples reflects recent advances in biotechnology development especially in the areas of genetic, infections and cancer diagnosis. The target DNA is detected by hybridization techniques derived from Southern's blotting. However such assays, based on the use of 32P labelled DNA probes, bring with them the associated problems of handling radioactive materials. In order to overcome these difficulties, a number of chemiluminescent detection methods have recently been developed. These new, alternative probe labelling procedures and chemiluminescent detection methods are easy to use in routine assays performed in research laboratories as well as for medical applications, and can reach the level of sensitivity found in classical radiolabelling techniques. The techniques investigated include peroxydase, biotin 16-dUTP or digoxigenin 11-dUTP probe labelling. The target DNAs are transferred onto nitrocellulose or nylon membranes and further fixed by heat or UV crosslinking. Specific hybridization on the target DNA is finally revealed by the use of chemiluminescent substrates. For all these techniques the detection limit is 10 aM (attomol) of a 561 bp target DNA. However for the probes labelled with peroxydase and with digoxigenin the detection limit drops to 1.0 aM of the target DNA. In the present paper we shall compare several of these DNA labelling and detection procedures and show that the detection threshold can vary by as much as a factor of 20 from method to method. This is the first time that various chemiluminescent methods for label and detection of DNA are compared and evaluated in order to determine the best protocol. PMID- 7494100 TI - The team approach to the design of computer-assisted learning packages in medicine. AB - Some of the didactic teaching of clinical medicine is now being replaced by self directed learning, sometimes in the form of computer-assisted learning (CAL). This represents a significant change in the way medical students are taught and has implications for all those involved in medical education. This article reviews the approach of one medical school to the introduction of CAL into the teaching of some aspects of clinical medicine. It emphasizes the importance of the centralization of resources and of a multidisciplinary team approach in the design of CAL packages. PMID- 7494101 TI - Digital video for networked CAL delivery. AB - Digital moving images (AVI format), were integrated into a specific computer assisted learning (CAL) application, providing a highly visual and interactive self-learning tool in an area of the medical curriculum which is difficult to teach by traditional methods. This paper outlines the process used to obtain the digital video using relatively inexpensive equipment and the subsequent delivery over a university network without resorting to major upgrading of the network specifications. PMID- 7494102 TI - An introduction to scriptwriting for video and multimedia. AB - The elements of audiovisual productions are explained and illustrated, including words, moving images, still images, graphics, narration, music, landscape sounds, pacing and tilting and font styles. Three different production styles are analysed, and examples of those styles are discussed. Rules for writing spoken words, composing blocks of information, and explaining technical information to a lay audience are also provided. Storyboard and scripting forms and examples are included. PMID- 7494103 TI - Teaching of human anatomy: a role for computer animation. AB - Computer-assisted learning fulfils an important need for pictorial representation of the functions of organs and systems. The various computer techniques of animation and morphing provide promising horizons for medical educational technology. Image acquisition is one of the most resource-intensive components of animation sequence development. Images can be drawn as originals or can be copied/scanned from various sources. By standardizing the initial (starting) image to the particular/basic need of the teacher and projecting the end-point image by using a vector animation package, 'films' can be created to demonstrate any form of movement. In the Anatomy Department, Sultan Qaboos University in Muscat, computer-animated tutorials are being introduced to illustrate normal and abnormal functional anatomy. The heart and its valve mechanisms have been selected as a pilot study. The student response is very positive and the technique has great potential. Embryology animations showing the formation and growth of organs such as the brain and spinal cord are also being developed. PMID- 7494104 TI - Medical education by satellite: the EuroTransMed experience. AB - Continuing medical education is a key feature of ensuring quality delivery of health care by doctors. In a rapidly changing technological environment it is important that medical education is undertaken in an effective and efficient manner. EuroTransMed, set up as an experiment to investigate the use of satellite delivered continuing medical education, now supplies regular programming to receive sites in 20 European countries. Programmes are broadcast live and are interactive, enabling doctors to have access to experts and opinion leaders and to question and challenge them. This review paper highlights the development and technical problems which have taken place during the initial establishment of EuroTransMed. The current status and future prospects are identified. PMID- 7494105 TI - Cell-mediated cytotoxicity in perforin-less mice. AB - We have used a perforin-less (PO) mouse to explore alternate CTL-mediated lytic pathways. PO mice are unable to overcome an infection with LCMV in vivo. Nevertheless, splenocytes from infected mice show vigorous, antigen-specific cytotoxicity that requires the presence of the Fas antigen on target cells. The Fas lytic pathway is virtually indistinguishable, in terms of kinetics and magnitude of cytotoxicity, from perforin/granzyme-mediated lysis. It is rapidly induced in CTL upon occupation of the TcR, and requires protein synthesis for full expression. Upon removal of the activating signal, the capacity for fas mediated lysis rapidly disappears. PO mice infected with LCMV also undergo what appears to be a CD8-mediated immunopathology, and rarely live beyond one month. The precise basis of this pathology is unknown at present. Given the widespread distribution of Fas in mice, particularly on inflamed tissues, the complete failure to clear virus from any tissue or organ is surprising. PMID- 7494106 TI - The function of the CD4 coreceptor in the development of T cells. AB - T cells with helper activity can be found in mice that lack expression of the CD4 glycoprotein. The CD4 promoter is active in these cells; they respond to antigens presented by MHC class II molecules; they do not express CD8 and they do not depend on MHC class I for their development. By such criteria, these CD8- T cells resemble normal CD4+ helper T cells. The development of the helper lineage in CD4 null mice can be potentiated by expression of transgenes that encode either wild type CD4, or a deletion mutant of CD4 that lacks the cytoplasmic tail and therefore cannot interact with the tyrosine kinase p56lck. These observations suggest that CD4 is not absolutely required for the specification of the helper cell lineage. The role of the CD4 molecule in the development of T cells and possible mechanisms by which it achieves its functions are discussed. PMID- 7494107 TI - T cell development in CD3-zeta mutant mice. AB - Increasing evidence points to multiple pathways of T lymphocyte development. The well characterized thymus-dependent pathway gives rise to T cells bearing TCR alpha beta heterodimers and either CD4 or CD8 alpha beta co-receptors. T cells of this lineage populate peripheral lymphoid compartments including lymph nodes, spleen, skin, and Peyer's patches. By comparison, factors which govern extrathymic T cell development are poorly understood. A variety of experiments have shown that intestinal intraepithelial lymphocytes (IELs) develop outside of the thymic environment, e.g., in the gut of nude, SCID, and beta 2m-/- mutant mice, and after transplanting bone marrow or fetal liver cells into irradiated thymectomized adult mice. This review focuses on the role of the CD3-zeta subunit in the development of both thymically and extrathymically derived T cells as determined by gene-targeting experiments in mice. Data from these and other T cell-related mutations continue to define crucial stages in thymocyte differentiation. Most interestingly, CD3-zeta mutant mice contain a unique population of intestinal IELs that develops independently of thymic selective processes and expresses a novel TCR/CD3 complex. PMID- 7494108 TI - Lymphocyte development and function in T-cell receptor and RAG-1 mutant mice. AB - We have used gene targeting in embryonic stem cells to generate mice with mutations in T-cell receptor (TCR) gene-alpha, TCR-beta, TCR-delta or the Recombination Activating Gene-1 (RAG-1). TCR-alpha or TCR-beta mutant mice are deficient in alpha beta T cells, but still contain gamma delta T cells and B cells. TCR-delta mutant mice are deficient in gamma delta T cells, but still contain alpha beta T cells and B cells. Mice doubly mutant for TCR-beta and TCR delta do not have any mature T cells, but still have B cells. RAG-1 mutant mice are totally deficient in both mature T cells and B cells. Here, I describe recent studies of thymocyte development in the mutant mice, and I review experiments addressing the function of the immune system of the mutant mice. PMID- 7494109 TI - Human antibodies from transgenic mice. AB - We have used homologous recombination in ES cells to engineer B cell-deficient mice that are incapable of expressing endogenous immunoglobulin heavy and kappa light chain genes. We find that B cell development in these mutant mice can be rescued by the introduction of human germline-configuration heavy- and kappa light-chain minilocus transgenes. The transgenes rearrange during B cell differentiation, and subsequently undergo class switching and somatic mutation in response to antigen stimulation; thus recapitulating both stages of the humoral immune response using human, rather than mouse, sequences. The mice can be immunized; and human sequence, antigen specific, monoclonal antibodies can be obtained using conventional rodent hybridoma technology. These animals are also of interest for studying the normal processes of immunoglobulin gene expression. We discuss the example of heavy chain class switching, which has not been previously observed within an autonomous transgene. PMID- 7494110 TI - Nonsurgical management of surgical otitis media with effusion. AB - The objective of this paper was to determine the effectiveness of combined steroid-antimicrobial therapy for otitis media with effusion (OME) of sufficient duration to justify tympanostomy tube insertion. A consecutive sample of 122 children with bilateral OME of at least three months duration, or unilateral OME of at least six months duration, despite treatment with one or more beta lactamase stable antibiotics was studied. The treatment group received prednisolone plus a beta-lactamase stable antibiotic for 10 days, with responders receiving an additional six weeks of chemoprophylaxis. The control group received no medication. The child's caregiver decided which group the child should be in. Resolution of effusion in all affected ears occurred in 32 per cent of steroid treated children and in 2 per cent of controls (p < 0.001) at three to four weeks post-therapy. Relapse of effusion occurred in over 40 per cent of initial responders within six months, reducing the final resolution rate to 25 per cent (95 per cent CI: 15-36 per cent). It was concluded that treatment with oral steroids should be considered in selected children with chronic OME prior to surgical intervention. One in every four children whose caregiver consents to this therapy may avoid or postpone surgery for at least six months. PMID- 7494111 TI - Management of retraction pockets of the pars tensa in children by excision and ventilation tube insertion. AB - Retraction pockets of the pars tensa in children can result in erosion of the ossicles leading to hearing loss and eventually cholesteatoma formation. Several different types of treatment for the more severe grades of retraction pocket have been described. The aim of this pilot study was to assess the outcome following simple excision and ventilation tube insertion of grade II, III and IV retraction pockets of the pars tensa. The eardrums were graded according to Sade's classification (1979). There were seven grade II and four grade III retractions. Ten eardrums healed completely in a mean time of 3.6 months (mean follow-up 16 months) and there was one residual perforation. Two retractions recurred and both of these were only grade I. Clinically, an improvement in hearing was reported in seven children (eight ears) and the average air conduction threshold gain for these patients was 16 dB. A larger prospective study is currently underway. PMID- 7494112 TI - A new incision for placement of cochlear implants. AB - A straight, vertical post-aural incision for the 'Cochlear' multichannel cochlear implant has been evaluated in 52 patients (20 adults and 32 children). Nineteen of the children were under three years of age and five of these were under two years of age. The 7 cm long incision is placed approximately 3 mm behind the post auricular crease and runs from the tip of the mastoid to a point 3 cm above the superior attachment of the pinna. The incision heals within several days. Because the incision is straight interruption of the blood supply to the flaps raised is the least possible. This also minimizes the possibility of scalp necrosis and implant extrusion. The likelihood of infection is reduced by the small size of the incision, minimal soft tissue dissection and small amount of dead space. Rapid healing has occurred in all cases despite infection in one. PMID- 7494113 TI - An approach to the audit of tinnitus management. AB - This study examines the advantages and internal consistency of an open-ended questionnaire (Benefit-problem questionnaire), over the Tinnitus questionnaire used by Jakes et al. (1985) in the auditing of a stress management programme. The Tinnitus questionnaire showed very little difference pre- and post-treatment, while the Benefit/problem questionnaire showed good improvement over a number of variables. This included some of the more traditional orthogonal values of tinnitus complaint. The clinical assessment made blindly and over a different time scale to the Benefit/problem questionnaire matched these positive results quite closely. PMID- 7494114 TI - Variation of expression of the gene for type 2 neurofibromatosis: absence of a gender effect on vestibular schwannomas, but confirmation of a preponderance of meningiomas in females. AB - Type 2 neurofibromatosis is a dominantly inherited disorder in which the great majority of sufferers develop bilateral vestibular schwannomas. In a UK study of 183 individuals from 112 families we have previously shown a fairly similar disease course within families, but quite marked inter-familial variation. We have confirmed an increase in severity when the gene is inherited from an affected mother, but evidence that women are more severely affected than men is lacking. Age at onset of symptoms, of deafness and at diagnosis are identical for the entire dataset and for a comparison of 10 male/female sibling pairs. Only three out of 42 pregnancies in symptomatic women were accompanied by a reversible worsening in symptoms due to vestibular schwannomas. Of 328 consecutive cases of unilateral vestibular schwannoma, there was no significant difference in the sex ratio or size. There now appears to be little evidence for a female hormonal effect on vestibular schwannomas. However, females with type 2 neurofibromatosis have significantly more meningiomas. PMID- 7494115 TI - Objective measurement of the benefit of walking sticks in peripheral vestibular balance disorders, using the Sway Weigh balance platform. AB - Following a lesion in the vestibular system visual, proprioceptive and residual vestibular information is integrated by the brain, to enable a patient to attain equilibrium. The basis of vestibular rehabilitation is to encourage these adaptive and compensatory mechanisms. Another form of rehabilitation is to provide some form of mechanical aid, and walking sticks are often used for this purpose in patients with balance disorders. There are no reported studies objectively assessing the use of walking sticks in patients with balance disorders. In this study we used the Sway Weigh balance platform (Raymar) to determine the efficacy of a walking stick in 25 patients with peripheral vestibular balance disorders. Patients were tested with their eyes opened and eyes closed whilst they were standing on a flat surface and on an air-filled bed (to alter limb proprioception) on the Sway Weigh balance platform. All the tests were carried out with, and without, a walking stick. The results demonstrate that a walking stick significantly reduces lateral body sway in patients with peripheral vestibular balance disorders. PMID- 7494116 TI - Topical active H1-antihistamines and their effect on nasal airway resistance. AB - The introduction of a topically active H1-antihistamine nasal spray Azelastine, has given an extra dimension in the management of allergic rhinitis. The drug acts rapidly and avoids the systemic adverse effects of antihistimines. An objective prospective study was performed to detect the effect of Azelastine nasal spray on nasal airway resistance. Twelve healthy adult volunteers with no rhinological problems were included in the study. Nasal cavities were sprayed with 280 micrograms (two puffs) of Azelastine nasal spray and the nasal airway resistance was measured with anterior rhinomanometry at intervals of 30 minutes for up to two hours. Our study has shown a statistically significant increase in the total nasal airway resistance following the use of Azelastine nasal spray in the absence of a subjective change in nasal airway resistance. There are substances when inhaled which can cause subjective improvement in nasal airway patency without changing the measured nasal airway resistance. However this medication gives no subjective change in nasal airway patency in spite of increasing nasal airway resistance. PMID- 7494117 TI - The value of radiological investigations in pre-endoscopic assessment of children with stridor. AB - The differential diagnosis of stridor in an infant depends on a careful history and examination, followed by radiological and endoscopic investigations. Currently a chest, lateral neck and antero-posterior, high kilovolt (Cincinnatti) view radiographs in association with a diagnostic barium swallow are performed prior to the definitive diagnostic procedure of microlaryngobronchoscopy. Our impression was that some of the routinely ordered radiological investigations were of limited value in the differential diagnosis. We undertook a retrospective audit study to determine the value of radiological investigations in the pre endoscopic assessment of infants with stridor. The radiological and endoscopic information of 100 infants presenting over the three-year period 1991-1993 at Great Ormond Street Hospital, London (a tertiary referral centre) was collected. It was found that only five out of 65 barium swallow investigations performed had consistent positive findings at diagnostic microlaryngobronchoscopy. The lateral neck and Cincinnatti views identified many of the more gross pathologies of the larynx and trachea e.g. the space occupying lesions, which occur infrequently. The more common diseases e.g. laryngomalacia or subglottic stenosis are rarely identified radiologically. Our results confirmed that radiology had a limited screening role and that in a child presenting with stridor the initial radiological assessment should be a chest radiograph with further imaging and a barium swallow only if an abnormality is found at microlaryngobronchoscopy. PMID- 7494118 TI - Punctate diathermy of the soft palate: a new approach in the surgical management of snoring. AB - We present our experiences in the surgical management of 21 snorers, utilizing a new approach designed to stiffen the palatal musculature, without compromising its function. This was achieved by using a punctate pattern of intrapalatal diathermy to achieve fibrosis within the palate and hence the requisite stiffening. The initial results of this procedure are encouraging showing a subjective improvement in snoring in 85.7 per cent of patients and an increase in sleeping in the same room as their partner every night, from 14.3 per cent pre operatively to 57.1 per cent post-operatively. PMID- 7494119 TI - Fine-needle aspiration cytology of submandibular gland lesions. AB - The usefulness of fine-needle aspiration cytology (FNAC) in the diagnosis and treatment of submandibular gland lesions is not well known. The 210 FNACs taken from submandibular gland lesions at Turku University Central Hospital between 1984 and 1991 were reviewed. Of these FNACs, 78 samples from primary lesions were confirmed histologically. Within this subset 10 FNACs were taken from benign neoplasms, all of which were correctly classified (sensitivity 100 per cent; specificity 88 per cent). Only four of the 14 FNACs from malignant lesions were cytologically considered malignant (sensitivity 29 per cent). On the other hand, four FNACs raised a false suspicion of malignancy (specificity 6 per cent). Out of 54 FNACs from non-neoplastic lesions 43 were correct (sensitivity 80 per cent; specificity 63 per cent). There were 104 patients (123 FNACs), who had not been operated on: the follow-up of these patients shows that in this subset of FNACs there were no false malignant but probably one false benign finding (1 per cent). We conclude that FNAC can offer valuable information about the type of the submandibular gland lesion, but the decision of operative and other treatment should not be based solely on the result of FNAC. PMID- 7494120 TI - Broncho-alveolar lavage in chronic upper respiratory tract infections. AB - The relationship between upper and lower respiratory tract infections has been demonstrated previously, although the effect of chronic infection of one tract on the other has not been well studied. This work analyses the broncho-alveolar lavage fluid of patients with chronic purulent rhino-sinusitis and reveals an increase in total and neutrophil cell counts as well as an increase in immuno globulin A levels. The associated increase in the neutrophil nitro-blue tetrazolium dye reduction test positivity provides evidence for increased phagocytosis to compensate for the increased contamination of the lower respiratory tract. PMID- 7494121 TI - The modified Valsalva manoeuvre to improve visualization of the hypopharynx during flexible nasopharyngoscopy. AB - A simple, safe and effective procedure for improving the diagnostic accuracy of nasopharyngoscopy is described. It is most useful for the exposure of the hypopharynx, especially in the elderly patient. PMID- 7494122 TI - Intrapetrous carotid artery aneurysm presenting as epistaxis and otalgia. AB - Aneurysm of the intrapetrous carotid artery is an extremely rare and potentially serious occurrence that presents diagnostic and therapeutic difficulties. Such aneurysms may follow trauma, atherosclerosis, mastoid surgery or most commonly can represent a developmental abnormality. We present the case of an 18-year-old female with a short history of recurrent left-sided otalgia and epistaxis who underwent successful endovascular balloon entrapment of a left intrapetrous carotid aneurysm. PMID- 7494123 TI - Enophthalmos secondary to maxillary sinus disease: single-stage operative management. AB - Maxillary sinus disease can lead to dehiscence of the orbital floor. Using endoscopic sinus surgery it is possible to simultaneously reconstruct the orbital floor and to assess and treat the underlying sinus disease. PMID- 7494124 TI - Adult floppy epiglottis: a simple surgical remedy. AB - Epiglottic prolapse during inspiration is an unusual cause of upper airway obstruction. We present a case of a previously fit, 73-year-old, male with upper airway obstruction secondary to a floppy epiglottis. We describe a simple method of treatment without the need for partial or total excision of the epiglottis or tracheostomy. PMID- 7494125 TI - Diet-modified sex hormone metabolism: is this the way forward in recurrent respiratory papillomatosis and squamous carcinoma prophylaxis? AB - We report two cases of recurrent laryngeal papillomatosis in women who subsequently developed squamous cell carcinoma as they approached the climacteric. The role of human papilloma virus (HPV) and oestrogen metabolism is outlined. The use of indoles from cruciferous vegetables is also discussed. PMID- 7494126 TI - Endoscopic balloon dilatation of subglottic stenosis. AB - Children with subglottic stenosis present a challenging problem to otolaryngologists. In many cases, a tracheostomy is necessary to safeguard the airway, but morbidity and mortality in the tracheostomized child may be significant. Therefore attempts to improve the airway by endoscopic means are often made; unfortunately, these have a variable success rate. Recent encouraging results in the use of balloon dilatation for subglottic stenosis led us to the successful use of this technique in a child whose stenosis had not responded to conventional endoscopic techniques. The reasons for the success are discussed. PMID- 7494127 TI - Primary tonsillar tuberculosis: a case report. AB - A 67-year-old man with an enlarged left tonsil underwent a tonsillectomy for the suspicion of malignancy. He had had a dental extraction six months earlier. Clinical and histopathological investigations established the diagnosis of primary tonsillar tuberculosis. PMID- 7494128 TI - Schwannoma of the posterior pharyngeal wall. AB - Schwannoma of the posterior pharyngeal wall, in a 36-year-old male patient is presented demonstrating the asymptomatic nature of the tumour (which is peculiar to schwannomas). The tumour gradually increased in size, destroyed the body of the third cervical vertebra but remained asymptomatic. The patient only presented to hospital when the tumour obstructed the airway, because of its large size. PMID- 7494129 TI - Recurrent cervical abscess due to piriform sinus fistula. AB - A case of recurrent abscess on the left anterior neck due to a left piriform sinus fistula is presented. It is known that a piriform sinus fistula may cause acute suppurative thyroiditis, and also that it induces the recurrent anterior cervical abscess described in this case. Here, we suggest that barium swallow is an important aid to diagnosis cases of recurrent cervical abscess. PMID- 7494130 TI - Pseudoaneurysm of the carotid artery with haemorrhage into the hypopharynx. AB - Pseudoaneurysms of the extracranial carotid arteries are rarely seen following irradiation for cancers of the head and neck. We present a patient with a pseudoaneurysm of the common carotid artery following a radical neck dissection and irradiation for thyroid carcinoma 20 years earlier. Following oesophagoscopical examination, a pseudoaneurysm of the right common carotid artery ruptured into the piriform sinus. The common carotid artery was embolized with multiple coils and the bleeding was halted. The relationship between the carotid artery aneurysm and irradiation, and the treatment of carotid artery aneurysm, is discussed. PMID- 7494131 TI - The management of benign thyroid goitre causing tracheo-oesophageal embarrassment. AB - The case of a massive benign thyroid goitre extending from the arch of aorta to the nasopharynx presenting with acute aerodigestive obstruction is reported. The management of the case is discussed and also a review of the relevant literature. PMID- 7494132 TI - Plasma cell granuloma of the supraglottic larynx. AB - The occurrence of plasma cell granuloma of the larynx appears to be unusual. Review of the literature revealed only two previously reported cases. We present an additional case of plasma cell granuloma of the larynx. The diagnosis was made by histological and immunohistochemical examinations. The tumour was successfully treated by radiation therapy. PMID- 7494134 TI - 186th Meeting of the Society for Endocrinology with the Endocrine Section of the Royal Society of Medicine. Abstracts. PMID- 7494133 TI - Unusual inflamed thyroglossal cyst. AB - The authors report a case of an unusual inflammatory reaction in a thyroglossal cyst. It consisted of broad papillary intraluminal projections covered by histiocytes and occasional multinucleate giant cells. This benign process should not be confused with a true papillary neoplasm, a rare complication of a thyroglossal cyst. PMID- 7494136 TI - Block by acetylcholine of mouse muscle nicotinic receptors, stably expressed in fibroblasts. AB - We have measured the concentration and voltage dependence of block by acetylcholine (ACh) of fetal- and adult-type mouse muscle nicotinic receptors, expressed in a fibroblast cell line. Data, obtained at a transmembrane potential of -60 mV and with ACh concentrations of 1 mM and above, are broadly consistent with the occlusion of an open channel with a single ACh+ ion (simple open channel block). The rate of recovery from block is approximately 40,000s-1 and has only a weak voltage dependence. This is in contrast to the strong voltage dependence observed for the degree of block. Deviations from the predictions of the simple model are seen in data collected at positive transmembrane potentials and at negative potentials for ACh concentrations < 1 mM. Less concentration dependence is observed than expected. Of a number of models tested, we demonstrate that two models incorporating both a high and a low affinity blocking site can predict our data. PMID- 7494137 TI - Regulation of the hyperpolarization-activated K+ channel in the lateral membrane of the cortical collecting duct. AB - An intermediate-conductance K+ channel (I.K.), the activity of which is increased by hyperpolarization, was previously identified in the lateral membrane of the cortical collecting duct (CCD) of the rat kidney (Wang, W. H., C. M. McNicholas, A. S. Segal, and G. Giebisch. 1994. American Journal of Physiology. 266:F813 F822). The biophysical properties and regulatory mechanisms of this K+ channel have been further investigated with patch clamp techniques in the present study. The slope conductance of the channel in inside-out patches was 50 pS with 140 mM KCl in the pipette and 5 mM KCl, 140 mM NaCl (NaCl Ringer's solution) in the bath. Replacement of the bath solution with symmetrical 140 mM KCl solution changed the slope conductance of the channel to 85 pS and shifted the reversal potential by 55 mV, indicating that the selectivity ratio of K+/Na+ was at least 10:1. Channel open probability (Po) in inside-out patches was 0.12 at 0 mV and was increased by hyperpolarization. The voltage-dependent Po was fitted with the Boltzmann's equation: Po = 1/[1 + exp(V-V1/2)zF/RT], with z = 1.2 and V1/2 = -40 mV. Addition of 2 mM tetraethylammonium or 500 mM quinidine to the bath blocked the activity of the K+ channel in inside-out patches. In addition, decrease in the bath pH from 7.40 to 6.70 reduced Po by 30%. Addition of the catalytic subunit of protein kinase A (PKAc; 20 U/ml) and 100 microM [corrected] MgATP to the bath increased Po from 0.12 to 0.49 at 0 mV and shifted the voltage dependence curve of channel activity toward more positive potentials by 40 mV. Two exponentials were required to fit both the open-time and the closed-time histograms. Addition of PKAc increased the long open-time constant and shortened the long closed-time constant. In conclusion, PKA-mediated phosphorylation plays an important role in the regulation of the voltage dependence of the hyperpolarization-activated K+ channel in the basolateral membrane of CCD. PMID- 7494135 TI - Intrinsic gating properties of a cloned G protein-activated inward rectifier K+ channel. AB - The voltage-, time-, and K(+)-dependent properties of a G protein-activated inwardly rectifying K+ channel (GIRK1/KGA/Kir3.1) cloned from rat atrium were studied in Xenopus oocytes under two-electrode voltage clamp. During maintained G protein activation and in the presence of high external K+ (VK = 0 mV), voltage jumps from VK to negative membrane potentials activated inward GIRK1 K+ currents with three distinct time-resolved current components. GIRK1 current activation consisted of an instantaneous component that was followed by two components with time constants tau f approximately 50 ms and tau s approximately 400 ms. These activation time constants were weakly voltage dependent, increasing approximately twofold with maximal hyperpolarization from VK. Voltage-dependent GIRK1 availability, revealed by tail currents at -80 mV after long prepulses, was greatest at potentials negative to VK and declined to a plateau of approximately half the maximal level at positive voltages. Voltage-dependent GIRK1 availability shifted with VK and was half maximal at VK -20 mV; the equivalent gating charge was approximately 1.6 e-. The voltage-dependent gating parameters of GIRK1 did not significantly differ for G protein activation by three heterologously expressed signaling pathways: m2 muscarinic receptors, serotonin 1A receptors, or G protein beta 1 gamma 2 subunits. Voltage dependence was also unaffected by agonist concentration. These results indicate that the voltage-dependent gating properties of GIRK1 are not due to extrinsic factors such as agonist-receptor interactions and G protein-channel coupling, but instead are analogous to the intrinsic gating behaviors of other inwardly rectifying K+ channels. PMID- 7494138 TI - A cellular mechanism for nitric oxide-mediated cholinergic control of mammalian heart rate. AB - The biochemical signaling pathways involved in nitric oxide (NO)-mediated cholinergic inhibition of L-type Ca2+ current (ICa[L]) were investigated in isolated primary pacemaker cells from the rabbit sinoatrial node (SAN) using the nystatin-perforated whole-cell voltage clamp technique. Carbamylcholine (CCh; 1 microM), a stable analogue of acetylcholine, significantly inhibited ICa(L) after it had been augmented by isoproterenol (ISO; 1 microM). CCh also activated an outward K+ current, IK(ACh). Both of these effects of CCh were blocked completely by atropine. Preincubation of the SAN cells with L-nitro-arginine methyl ester (L NAME; 0.2-1 mM), which inhibits NO synthase (NOS), abolished the CCh-induced attenuation of ICa(L) but had no effect on IK(ACh). Coincubation of cells with both L-NAME and the endogenous substrate of NOS, L-arginine (1 nM), restored the CCh-induced attenuation of ICa(L), indicating that L-NAME did not directly interfere with the muscarinic action of CCh on ICa(L). In the presence of ISO the CCh-induced inhibition of ICa(L) could be mimicked by the NO donor 3-morpholino sydnonimine (SIN-1; 0.1 mM). SIN-1 had no effect on its own or after a maximal effect of CCh had developed, indicating that it does not inhibit ICa(L) directly. SIN-1 failed to activate IK(ACh), demonstrating that it did not activate muscarinic receptors. Both CCh and NO are known to activate guanylyl cyclase and elevate intracellular cGMP. External application of methylene blue (10 microM), which interferes with the ability of NO to activate guanylyl cyclase, blocked the CCh-induced attenuation of ICa(L). However, it also blocked the activation of IK(ACh), suggesting an additional effect on muscarinic receptors or G proteins. To address this, a separate series of experiments was performed using conventional whole-cell recordings with methylene blue in the pipette. Under these conditions, the CCh-induced attenuation of ICa(L) was blocked, but the activation of IK(ACh) was still observed. Methylene blue also blocked the SIN-1 induced decrease in ICa(L). 6-anilino-5,8-quinolinedione (LY83583; 30 microM), an agent known to decrease both basal and CCh-stimulated cGMP levels, prevented the inhibitory effects of both CCh and SIN-1 on ICa(L), but had no effect on the activation of IK(ACh) by CCh. In combination, these results show that CCh- and NO induced inhibition of ICa(L) is mediated by cGMP.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 7494140 TI - The mammalian Na+/H+ antiporters NHE-1, NHE-2, and NHE-3 are electroneutral and voltage independent, but can couple to an H+ conductance. AB - Na+/H+ exchange in vertebrates is thought to be electroneutral and insensitive to the membrane voltage. This basic concept has been challenged by recent reports of antiport-associated currents in the turtle colon epithelium (Post and Dawson, 1992, 1994). To determine the electrogenicity of mammalian antiporters, we used the whole-cell patch clamp technique combined with microfluorimetric measurements of intracellular pH (pHi). In murine macrophages, which were found by RT-PCR to express the NHE-1 isoform of the antiporter, reverse (intracellular Na(+)-driven) Na+/H+ exchange caused a cytosolic acidification and activated an outward current, whereas forward (extracellular Na(+)-driven) exchange produced a cytosolic alkalinization and reduced a basal outward current. The currents mirrored the changes in pHi, were strictly dependent on the presence of a Na+ gradient and were reversibly blocked by amiloride. However, the currents were seemingly not carried by the Na+/H+ exchanger itself, but were instead due to a shift in the voltage dependence of a preexisting H+ conductance. This was supported by measurements of the reversal potential (Erev) of tail currents, which identified H+ (equivalents) as the charge carrier. During Na+/H+ exchange, Erev changed along with the measured changes in pHi (by 60-69 mV/pH). Moreover, the current and Na+/H+ exchange could be dissociated. Zn2+, which inhibits the H+ conductance, reversibly blocked the currents without altering Na+/H+ exchange. In Chinese hamster ovary (CHO) cells, which lack the H+ conductance, Na+/H+ exchange produced pHi changes that were not accompanied by transmembrane currents. Similar results were obtained in CHO cells transfected with either the NHE-1, NHE-2, or NHE-3 isoforms of the antiporter, indicating that exchange through these isoforms is electroneutral. In all the isoforms tested, the amplitude and time-course of the antiport-induced pHi changes were independent of the holding voltage. We conclude that mammalian NHE-1, NHE-2, and NHE-3 are electroneutral and voltage independent. In cells endowed with a pH-sensitive H+ conductance, such as macrophages, activation of Na(+)-H+ exchange can modulate a transmembrane H+ current. The currents reported in turtle colon might be due to a similar "cross talk" between the antiporter and a H+ conductance. PMID- 7494141 TI - The search for a taxol-producing microorganism among the endophytic fungi of the Pacific yew, Taxus brevifolia. AB - Endophytic microbes associated with the Pacific yew tree, Taxus brevifolia, were examined as potential sources of the anticancer drug taxol [1], a secondary metabolite of the host organism. The first promising organism found was the novel fungus, Taxomyces andreanae, which was isolated from the inner bark of a yew tree growing in northwestern Montana. It appears to produce taxol and other taxanes in de novo fashion when grown in semi-synthetic liquid media. The presence of 1 in the fungal extract was confirmed by mass spectrometry, comparative chromatographic behavior with "yew" taxol, reactivity with taxol-specific monoclonal antibodies, and 9KB cytotoxicity studies. Both acetate-1-14C and phenylalanine UL-14C served as precursors of taxol-14C in fungal culture labeling studies, confirming the de novo synthesis of 1 by the fungus. Immunoassay techniques are currently being used to screen extracts of Taxomyces andreanae for new taxanes, and to determine if other endophytic fungi are taxol producers. PMID- 7494139 TI - The relationship between membrane fluidity and permeabilities to water, solutes, ammonia, and protons. AB - Several barrier epithelia such as renal collecting duct, urinary bladder, and gastric mucosa maintain high osmotic pH and solute gradients between body compartments and the blood by means of apical membranes of exceptionally low permeabilities. Although the mechanisms underlying these low permeabilities have been only poorly defined, low fluidity of the apical membrane has been postulated. The solubility diffusion model predicts that lower membrane fluidity will reduce permeability by reducing the ability of permeant molecules to diffuse through the lipid bilayer. However, little data compare membrane fluidity with permeability properties, and it is unclear whether fluidity determines permeability to all, or only some substances. We therefore studied the permeabilities of a series of artificial large unilamellar vesicles (LUV) of eight different compositions, exhibiting a range of fluidities encountered in biological membranes. Cholesterol and sphingomyelin content and acyl chain saturation were varied to create a range of fluidities. LUV anisotropy was measured as steady state fluorescence polarization of the lipophilic probe DPH. LUV permeabilities were determined by monitoring concentration-dependent or pH sensitive quenching of entrapped carboxyfluorescein on a stopped-flow fluorimeter. The relation between DPH anisotropy and permeability to water, urea, acetamide, and NH3 was well fit in each instance by single exponential functions (r > 0.96), with lower fluidity corresponding to lower permeability. By contrast, proton permeability correlated only weakly with fluidity. We conclude that membrane fluidity determines permeability to most nonionic substances and that transmembrane proton flux occurs in a manner distinct from flux of other substances. PMID- 7494142 TI - Natural product drug discovery and development: new perspectives on international collaboration. AB - Until recently, the prevailing attitude in developed nations regarded the world's genetic resources, which are mainly concentrated in the developing world, as a common resource of humankind, to be exploited freely irrespective of national origin. With the devastation being wreaked in the tropical rainforests and the resurgence in interest in recent years in the discovery of novel drugs from natural sources, particularly plants and marine organisms, the international scientific community has realized that the conservation of these global genetic resources and the indigenous knowledge associated with their use are of primary importance if their potential is to be fully explored. With this realization has come a recognition that these goals must be achieved through collaboration with, and fair and equitable compensation of, the scientists and communities of the genetically rich source countries. The signing of the United Nations Convention on Biological Diversity by nearly all of the World's nations has emphasized the need for the implementation of such policies. In this review, the articles of the Convention of relevance to the activities and practices of the natural products scientific community are briefly discussed. This discussion is followed by a summary of policies for international collaboration and compensation being implemented by several developed country organizations, and the perspectives on the current developments given by representatives of some of the source countries located in the regions of greatest biodiversity. PMID- 7494143 TI - Effect of anthraquinone derivatives on lipid peroxidation in rat heart mitochondria: structure-activity relationship. AB - Lipid peroxidation was induced in rat heart mitochondria with FeSO4 and the inhibitory effects of various anthraquinone derivatives were compared. Oxygen consumption and malondialdehyde formation were used to quantitate the amount of lipid peroxidation. Emodin [2], alizarin [13], and alizarin complexone [14] significantly inhibited lipid peroxidation; their potency as inhibitors of lipid peroxidation was higher than that of alpha-tocopherol. Structure-activity analysis showed that two hydroxyl groups arranged in either the ortho- or meta position in the C ring of the anthraquinone nucleus are required for such derivatives to inhibit lipid peroxidation. The diphenyl-p-picrylhydrazyl test showed that alizarin [13] and alizarin complexone [14] are free-radical scavengers while emodin [2] is not. The mechanism for emodin [2] to inhibit lipid peroxidation is most likely due to inhibition on the propagation of lipid peroxyl radicals in the mitochondrial membrane. PMID- 7494144 TI - Anti-AIDS agents, 21. Triterpenoid saponins as anti-HIV principles from fruits of Gleditsia japonica and Gymnocladus chinensis, and a structure-activity correlation. AB - Gleditsia saponin C [1] and gymnocladus saponin G [2] were isolated from Gleditsia japonica and Gymnocladus chinensis, respectively, as anti-HIV principles. Compounds 1 and 2 demonstrated inhibitory effects against HIV replication in H-9 cells with EC50 values of 1.1 and 2.7 microM, respectively. Evaluation of the anti-HIV activities of the prosapogenins of 1 and 2 suggested that the unusual monoterpenyl moieties in 1 and 2 are essential for their anti HIV activity. Derivatives of echinocystic acid [8], the aglycone of compound 1, were also prepared and evaluated for inhibitory activity against HIV replication. 3,16-Di-O-acetylechinocystic acid [12] was shown to be an anti-HIV agent with an EC50 value of 2.3 microM. PMID- 7494146 TI - Identification of an N-acetyl keto derivative of fumonisin B1 in corn cultures of Fusarium proliferatum. AB - A method is presented for the separation and identification of a new N-acetyl keto derivative of fumonisin B1 (FB1) produced in solid corn culture. Cultures of Fusarium proliferatum (M-1597) were purified using preparative hplc, and the new fumonisin was detected by negative-ion esms. Structures were confirmed by 1H- and 13C-nmr spectroscopy. The new fumonisin differs from FB1 in that the tricarballylic acid functionality at the C-15 position of the eicosane backbone is replaced by a ketone and the amino group is acetylated. Direct analysis of the culture material by negative-ion electrospray lc/ms confirmed that the new fumonisin is produced naturally by the fungus. PMID- 7494145 TI - Enzyme inhibitors: new and known polybrominated phenols and diphenyl ethers from four Indo-Pacific Dysidea sponges. AB - Extracts and pure compounds isolated from four samples of Dysidea sp. sponges collected from two geographically distinct regions of the Indo-Pacific (Chuuk Atoll and Fiji) were assayed against five different enzyme assays, four of which are relevant to anticancer drug discovery and one of which (15-lipoxygenase) may detect compounds significant in modulating the development of atherosclerotic plaque. The pure compounds that inhibited various enzymes were polybrominated phenols and polybrominated phenoxyphenols. Fourteen of these phenols were isolated, six of which were new compounds. A variety of the phenols inhibited inosine monophosphate dehydrogenase (IMPDH), guanosine monophosphate synthetase, and 15-lipoxygenase. No activity was observed with protein tyrosine kinase pp60v src or matrix metalloprotease. PMID- 7494147 TI - Longicin and goniothalamicinone: novel bioactive monotetrahydrofuran acetogenins from Asimina longifolia. AB - Longicin [1] and (2,4-cis and trans)-goniothalamicinone [2], two new monotetrahydrofuran Annonaceous acetogenins, have been isolated from the leaves and twigs of Asimina longifolia (the long leaf paw paw) by the use of the brine shrimp lethality test for bioactivity-directed fractionation. The structures were elucidated based on spectroscopic and chemical methods. Compound 1 was converted to its ketolactone isomer, (2,4-cis and trans)-longicinone [3], to aid the stereochemical elucidation of 1. Compounds 1-3 showed selective and potent cytotoxicities to certain human tumor cell lines, with the potency of 1 against pancreatic carcinoma (PaCa-2) over one million times that of adriamycin. Nine known cytotoxic acetogenins, annonacin, xylomaticin, isoannonacin, gigantetrocins A and B, muricatetrocins A and B, gigantetrocin-A-one and goniothalamicin, were also isolated for the first time from this species. PMID- 7494148 TI - Amoenolide K and amoenolide K 19-acetate, two grindelane peroxides from Amphiachyris amoena. Isolation, structure determination, and preparation of amoenolide K from amoenolide A by photochemical oxygenation. AB - Two new 9,13-diepoxy labdane diterpenes, amoenolide K [1] and its 19-acetate [2], were isolated from the aerial parts of the composite Amphiachyris amoena and their structures were established by spectral methods, especially high-field nmr spectroscopy. Amoenolide K [1] was prepared from amoenolide A [3] by singlet oxygen addition via the ene reaction. A study was made of the ene reaction products with amoenolide K triacetate [5] which showed them to arise from stereospecific oxygen addition to the 8,9-double bond, with the exception of the sterically hindered beta-side at C-9, for which no products were isolated. PMID- 7494149 TI - New antifungal bithienylacetylenes from Blumea obliqua. AB - Two new bioactive bithienylacetylenes isolated from Blumea obliqua have been characterized as 5'-methyl-5-[4-(3-methyl-1-oxobutoxy)-1-butynyl]-2,2'- bithiophene [1] and 5'-hydroxymethyl-5-[butyl-3-en-1-yn]-2,2'-bithiophene isovaleroxy ester [2], respectively. Compounds 1 and 2 showed antifungal activity against Epidermophyton floccosum and Pleurotus ostreatus. PMID- 7494150 TI - Additional bioactive acetogenins, annomutacin and (2,4-trans and cis)-10R annonacin-A-ones, from the leaves of Annona muricata. AB - In a continuation of our research on bioactive components from the leaves of Annona muricata, three novel monotetrahydrofuran Annonaceous acetogenins, namely, annomutacin [1], (2,4-trans)-10R-annonacin-A-one [2], and (2,4-cis)-10R- annonacin-A-one [3], have been identified. Their structures were deduced by ms, nmr, ir, and uv spectral and chemical methods, and the absolute configurations were determined by Mosher ester methodology. A known bioactive amide, N-p coumaroyl tyramine, was also found. Compound 1 and the mixture of compounds 2 and 3 showed selective cytotoxicities against the human A-549 lung tumor cell line. PMID- 7494151 TI - New azacyclopropene derivatives from Dysidea fragilis collected in Pohnpei. AB - The sponge Dysidea fragilis from Pohnpei contained four azacyclopropene derivatives, (4E)-S-dysidazirine [2], which is the optical enantiomer of the known compound dysidazirine [1], (4Z)-dysidazirine [3], (4E)-S-antazirine [4], and (4Z)-antazirine [5]. The structures of the new compounds were elucidated by interpretation of spectral data. PMID- 7494152 TI - Anguillosporal, a new antibacterial and antifungal metabolite from the freshwater fungus Anguillospora longissima. AB - Anguillosporal [1], a new antifungal and antibacterial metabolite, has been isolated from the freshwater fungus Anguillospora longissima (CS-869-1A). Its structure was determined as 2,4-dihydroxy-3-ethyl-6-(1'-methylpentyl) benzaldehyde on the basis of ms, 1H-nmr, and 13C-nmr data. PMID- 7494153 TI - Phase transfer and point-spread function of the human eye determined by a new asymmetric double-pass method. AB - A recent study has shown that the double-pass method provides a good estimate of the ocular modulation transfer function (MTF) but that it does not yield the phase transfer function (PTF) [J. Opt. Soc. Am. A 12, 195 (1995)]. Therefore, one cannot recover the true retinal point-spread function (PSF). We present a modification of the double-pass method to overcome this problem. The key is to break the symmetry between the two passes. By using an unexpanded Gaussian input beam, we produce a diffraction-limited PSF for the first passes. Then, by using a large exit pupil, we get an aberrated PSF for the second pass. The double-pass aerial image is the cross correlation of both PSF's, so that the Fourier transform of such an aerial image directly provides the true retinal PTF, up to the cutoff frequency of the effective (small), diffraction-limited entrance pupil. The resulting double-pass aerial image is a blurred version of the true retinal PSF. Thus it shows the effect not only of even symmetric aberrations but also of odd and irregular aberrations such as coma. We have explored two different ways to retrieve the true retinal PSF: (a) deblurring of the aerial image and (b) PSF reconstruction combining PTF data with conventional double-pass MTF. We present promising initial results with both artificial and real eyes. PMID- 7494154 TI - Boundary conditions for diffusion of light. AB - In connection with recent work on remote imaging of random media by light, a straightforward generalization of the proper diffusion boundary conditions is presented that takes into account Fresnel reflection. The Milne problem at exterior boundaries is solved for various values of index of refraction, absorption, and scattering anisotropy parameters to yield extrapolated end points and extrapolation distances. A generalized interface condition is derived to replace the usual condition of continuity of intensity. Benchmark-quality numerical results are given for the extrapolation distance and for the new index dependent parameter in the interface conditions. Difficulties in using the extrapolated end point when the index is sufficiently large are discussed, and a new image procedure suitable for this case is presented. PMID- 7494155 TI - Posterior chamber intraocular lenses: long-term results in pediatric cataract patients. AB - Major problems in pediatric cataract patients include noncompliance with contact lenses resulting in amblyopia. Advances in intraocular cataract surgery have provided a better environment to perform intraocular lens (IOL) implantation in children. We prospectively analyzed the results of 34 consecutive pediatric patients who underwent cataract removal and insertion of an IOL. Operative technique performed was posterior scleral beveled or frown incision and IOL endocapsular fixation. Subgroups included 10 eyes with traumatic cataract, and 24 eyes with developmental cataracts. Six patients had bilateral IOLs. Data presented in each subgroup included initial and final visual acuity, age, sex, type of cataract, A and B scan biometry, early and late postoperative complications, time of YAG capsulotomy, postoperative refractive correction, and state of binocular vision. Preoperative visual acuity ranged from 20/70 to light perception. The success rate for postoperative visual acuity of 20/40 or better occurred in 29 of 34 eyes, or 85.3%. In the traumatic cataracts, 8 of 10 eyes (80%) obtained 20/40 vision or better. In developmental cataracts, 21 of 24 eyes (88%) achieved visual acuity of 20/40 or better. Amblyopia (three patients) or macular scar (two patients) accounted for reduced visual acuity in the five eyes with vision less than 20/40. Early complications included posterior synechiae and lens deposits. The primary late complication was opacification of the posterior capsule in 18 of 34 eyes. The average time for YAG capsulotomy post-cataract removal was 17 months. None of the 28 patients (34 eyes) developed glaucoma, IOL dislocation, or other significant postoperative problems related to IOL insertion.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7494156 TI - Haptic breakage following neonatal IOL implantation in a nonhuman primate model. AB - To evaluate the long-term safety of implanting intraocular lenses (IOLs) in neonatal eyes using a nonhuman primate model, a one-piece all polymethylmethacrylate (PMMA) IOL was implanted into the posterior chamber of 21 neonatal Rhesus monkey eyes following a lensectomy and anterior vitrectomy. These monkeys then were monitored for 26 to 40 months for complications. The eyes from seven monkeys were studied histopathologically. One or both haptics broke on 7 of the 21 (33%) IOLs in a mean of 21 months (range 6 to 31 months) after their implantation. Only one of the haptics from these seven eyes was found to be in the capsular bag when these eyes were studied histopathologically. Nine haptics had eroded into the iris leaflets, two into the ciliary body, and one into the anterior chamber. One additional haptic was in the sulcus. The haptics of one piece all-PMMA IOLs may break if implanted in neonatal eyes. A number of factors likely contributed to haptic breakage in these eyes, including an increased number of reoperations, malpositioning of IOL haptics, rapid circumferential and axial growth, and the formation of large Soemmerring's rings. PMID- 7494157 TI - Alignment success following medical rectus recessions in normal and delayed children. AB - Others have suggested that developmentally delayed children, who are frequently esotropic, have a poor prognosis following esotropia surgery. To date, no comparison of success rates in normal and delayed children following similar surgery has been made. We compared our long-term results following graded bilateral medial rectus recessions in consecutive normal (n = 62) and delayed (n = 29) children. We defined success as alignment maintained within 10 delta of orthophoria. Survival curves were compared using the Mantel-Haenszel statistic. Delayed children had a significantly poorer outcome (p = .05) throughout follow up, which ranged from 12 to 120 months (mean = 24 months). Early in the period of study, we observed a large proportion of overcorrections among the delayed children. Subsequent modification in the amount of surgery performed appeared to improve the initial and long-term success in these children. We conclude that delayed children do have a poorer prognosis than normal children following medial rectus recession. This prognosis may be improved by more conservative amounts of surgery. PMID- 7494158 TI - Dissociated horizontal deviation and accommodative esotropia: treatment options when an eso- and an exodeviation co-exist. AB - Dissociated strabismus includes movements in the vertical, horizontal, and torsional axes. Dissociated vertical deviation (DVD) is an oversimplified label because it implies a purely vertical deviation. When the abducting component of the dissociated strabismus complex is prominent, the term dissociated horizontal deviation (DHD) is used. We present six patients with accommodative esotropia who developed prominent DHD. Only two patients have a prior history of congenital esotropia. Latent nystagmus was observed in five of the patients. With the combination of DHD and esotropia, an esodeviation during active fixation may become an exodeviation when the patient is visually inattentive. Five patients required surgery for DHD. Four options are used and discussed. DHD can be treated alone with lateral rectus muscle recession. DHD can be treated as above, but combined with medial rectus muscle recessions or posterior fixation sutures to simultaneously treat the esotropia. The esotropia can be treated alone, possibly with a reduced surgical dosage. If the patient varies from an esodeviation to an exodeviation, but is straight much of the day, observation without surgical therapy may be advisable. Just as a hypotropia can have an overlying DVD, an esotropia can have an overlying DHD. This combination may masquerade as variable angle esotropia or may present as an esodeviation when the patient is visually attentive and an exodeviation when the patient is visually inattentive. PMID- 7494159 TI - Random dot stereoacuity following surgical correction of infantile esotropia. AB - Surgical correction of infantile esotropia prior to age 2 is associated with a higher prevalence of fusion and stereopsis than surgical correction after 2 years of age. The advantages and disadvantages of surgical intervention at the early or late end of this window have been debated in the literature. In the present study, random dot (RD) stereoacuity outcomes were evaluated in order to determine whether a binocular sensory benefit is associated with early or late surgery. Participants were 73 healthy children enrolled in a prospective study of visual development in infantile esotropia. All children had initial surgical correction at 5 to 16 months of age. RD stereoacuity was evaluated at approximately 5 years of age (59.7 +/- 14.9 months). Overall, 41.1% of children demonstrated RD stereopsis. The percentage of children demonstrating RD stereopsis was not significantly different among groups that were surgically corrected at 5 to 8 months (43.8%), 9 to 12 months (47.4%), and 13 to 16 months (31.8%). However, among those children who achieved RD stereopsis, the prevalence of foveal (< 60 sec) or macular (61 to 200 sec) stereoacuity was significantly higher among those who had surgery at 5 to 8 months (42.9%; Z = 2.06, p < 0.02) or 9 to 12 months (55.6%; Z = 2.38, p < 0.009) than among those who had surgery at 13 to 16 months (0.0%). Although surgical correction of infantile esotropia during the first year of life is not associated with a higher prevalence of RD stereopsis, it is associated with better RD stereoacuity among those children who achieve stereopsis following surgery. PMID- 7494160 TI - Development of compensating exotropia with anomalous retinal correspondence after early infancy in congenital homonymous hemianopia. PMID- 7494161 TI - Hydroxypropylmethylcellulose and sodium hyaluronate in adjustable strabismus surgery. AB - The efficacy of hydroxypropylmethylcellulose and sodium hyaluronate as adjuncts during adjustable strabismus surgery was evaluated experimentally. Fifteen randomized NZA rabbits underwent a bilateral 4-mm recession of the superior rectus followed by a 0.05 mL injection of submuscular hydroxypropylmethylcellulose 2% (HPMC) in one eye and sodium hyaluronate 1% (Healon) in the other eye. Five additional rabbits (10 eyes), used as controls, underwent the same procedure, but no viscoelastic was injected after surgery. The force needed to move the muscle 4 mm toward its original insertion was measured immediately and 24 hours after the operation, using a force gauge. The force needed to adjust the muscles increased 326.0%, 98%, and 124.7%, respectively in the control, Healon and HPMC groups (p < 0.001). No significant difference was found between the two viscoelastic substances. Histologic evaluation of the operation sites disclosed similar inflammation in both controls and viscoelastic groups (p = .5960). PMID- 7494162 TI - Brown's syndrome with cyclic characteristic: case report and review of physiopathologic mechanism. AB - This article reports on an acquired Brown's syndrome case presenting with cyclic characteristics. Restriction of elevation in adduction was observed in the right eye of a 15-year-old boy who had been referred with a complaint of diplopia that occurred on waking in the morning and continuing until noon. The same situation was noted following afternoon naps. The patient was able to improve his complaint by performing a forced elevation in adduction while massaging his right trochlear region and the restriction did not recur until the next morning. PMID- 7494163 TI - Glaucoma and findings simulating glaucoma in the Rubinstein-Taybi syndrome. AB - Information is reviewed on the ophthalmologic findings in 614 individuals with Rubinstein-Taybi syndrome (RTS). The data were collected from the world literature, from communication with colleagues and with families of individuals with RTS, and from personal observations. Particular emphasis is given in this article to the association of RTS with glaucoma and five other findings that may be confused with glaucoma (corneal lesions, megalocornea, colobomatous or cystic optic nerve, excavation of papilla, and large cup-to-disc ratio). A case report is presented including autopsy results on a 5-year-old black female with RTS, corneal lesions, colobomas of the optic nerves, and normal intraocular pressure. PMID- 7494164 TI - Retinal detachment in a child as the first sign of leukemic relapse: histopathology, MRI findings, treatment, and tumor-free follow up. AB - Improved leukemia therapies in children have brought about prolonged remissions with extramedullary relapses being reported in sites other than the most common (bone marrow, testes, brain, and spinal cord). A 3-1/2 year-old boy with a history of acute lymphocytic leukemia presented with total retinal detachment in one eye. Painful glaucoma unresponsive to medical therapy necessitated enucleation. Histopathologic examination documented the presence of a dense leukemic cellular infiltrate replacing a totally detached, necrotic retina. Tumor cells also were present in the optic nerve. The child had remained free of leukemia for 3 years after systemic and intrathecal chemotherapy, supplemented by craniospinal radiation. This represents the first case of relapse of acute lymphocytic leukemia presenting solely as a retinal detachment. Our case also underscores the point that the treatment of leukemia after an isolated ocular relapse can be associated with a favorable outcome. PMID- 7494165 TI - Modified periodontal probe for simplified forced duction measurements. AB - A periodontal probe has been modified for use at surgery as a quantitative length tension measuring device. It is simple and rapid to use, as well as inexpensive. In addition to zero length and tension, readings can be made at 25 g and 30 g of tension. Results of the test are described in four patients with restrictive or paralytic strabismus. PMID- 7494166 TI - Pilomatrixoma of the eyelid. PMID- 7494167 TI - Atypical presentation of orbital rhabdomyosarcoma: a report of two cases. PMID- 7494168 TI - Coloboma associated with Rubinstein-Taybi syndrome. PMID- 7494169 TI - Nasolacrimal duct obstruction study. PMID- 7494170 TI - Nasolacrimal duct obstruction study. PMID- 7494172 TI - Migration and proliferation of progenitor cells in the connective tissue of rat gingival papilla. AB - The aim of this investigation was to determine whether gingival fibroblast progenitor cells undergo division and whether there is evidence of migration of their progeny from the paravascular sites within the interdental gingival septum (IGS). 30 male hooded Lister rats aged 6 weeks, were killed in groups of ten, 3, 75 and 171 h after a single injection of tritiated thymidine. Autoradiographs were examined of transverse Historesin sections of the papilla between second and third mandibular molars in 29 specimens, taken from equidistant intervals between the col and alveolar bone crest. Median grain counts in the whole papilla were greatest within 10 microns of blood vessels at 3 h (p < 0.02) and decreased throughout the papilla with time (p < 0.0001). The median grain counts at level 5, nearest the periodontal ligament, were greatest at all times (p < 0.01). The distance of labelled nuclei from blood vessel walls increased with time, most notably at level 5 (p < 0.001). The mean Clustering Index increased (p < 0.01) from 4.6% at 3 h to 12.3% and 11.6% at 75 h and 171 h respectively. At 75 h, the median grain count of clustered nuclei was smaller (p < 0.01) than that of non clustered nuclei. These data are consistent with the occurrence of clonal division and migration within the IGS and the presence of a slowly cycling cell population near to the periodontal ligament. PMID- 7494171 TI - Response of human gingival fibroblasts to prostaglandins. AB - The effects of various prostaglandins (PGs) on the functions of human gingival fibroblasts (Gin-1 cells; ATCC CRL 1292) were examined by phase-contrast microscopy, cell-counting and radioautographic experiments. Tested PGs were PGA1, PGA2, PGB1, PGB2, PGD2, PGE1, PGE2, PGF1 alpha, PGF2 alpha, PGI2, 6-keto-PGF1 alpha, 9 alpha-11 alpha-methanoepoxy-PGF2 alpha, and thromboxane (TX) B2. PGA1 and PGD2 at 30 microM caused morphological deformation of Gin-1 cells. All the PGs tested at 30 microM suppressed the proliferation of Gin-1 cells in the logarithmic growth phase. Furthermore, all the PGs tested at 10 microM suppressed DNA synthesis, collagen synthesis, and noncollagenous protein synthesis in confluent Gin-1 cells, while exerting no effect on GAG synthesis. The concentrations of PGs used are beyond those found in healthy gingiva. However, in periodontitis the local concentrations of some PGs within the gingiva are expected to be extremely elevated beyond the physiological level. These results suggest that PGs may play an important role as a negative regulator in metabolism and some pathologic gingival conditions by suppressing the functions of gingival fibroblasts. PMID- 7494173 TI - Residual effect of chlorhexidine gluconate in 4-day plaque regrowth crossover trials, and its implications for study design. AB - The efficacy of oral hygiene agents in preventing the accumulation of plaque from a zero baseline is conveniently evaluated in the 4-day plaque regrowth model. In many studies a panel of volunteers has been used to compare several agents according to a highly efficient Latin square based crossover design. Often chlorhexidine gluconate is included as a positive control; there is concern that the effect of this potent agent might persist for longer than the 3-day washout period often employed. To test this hypothesis of residual effect, we put together the evidence relating to possible carryover from 12 studies. The residual effect of chlorhexidine differs from that of an inert rinse such as saline or water. Consequently, a longer washout period, such as 10 days, is preferable. Studies omitting chlorhexidine are worth considering. Designs should be balanced for residual effects, at least from the preceding treatment period; several useful designs with this property are presented. PMID- 7494174 TI - Microflora in adult periodontitis. AB - The subgingival microflora of adult periodontitis was studied in 8 adults (36-47 years) and compared with that of 10 periodontally healthy individuals (36-43 years). A total of 64 periodontal lesions were examined, and classified according to the attachment level in three categories: attachment loss > 6 mm, attachment loss 4-6 mm and attachment loss < 4 mm. Also for comparative purposes 20 gingival sulci were evaluated. Samples were taken using three standardized paper points and were incubated anaerobically in selective and non-selective media. The results showed a statistically significant association of Capnocytophaga gingivalis and Capnocytophaga sputigena with moderate periodontal lesions, while Haemophilus segnis has been correlated to severe periodontal lesions. We concluded that C. gingivalis, C. sputigena and H. segnis might be potentially conductive to periodontal deterioration in adult periodontitis. PMID- 7494175 TI - Distribution of Porphyromonas gingivalis and Treponema denticola in human subgingival plaque at different periodontal pocket depths examined by immunohistochemical methods. AB - Localization of Porphyromonas gingivalis and Treponema denticola in different areas of subgingival plaque from advanced adult periodontitis patients was studied immunohistochemically using sensitive immunogold-silver staining and immunoelectron microscopy. Fourteen periodontally diseased teeth were extracted without damaging the subgingival plaque, fixed, and embedded. The subgingival plaque samples were sectioned according to four different pocket depths (0-2 mm, 2-4 mm, 4-6 mm and > or = 6 mm). Serial thin sections were stained using specific antisera to P. gingivalis or T. denticola and then with secondary antibody labelled with colloidal-gold. Cells of both P. gingivalis and T. denticola were predominantly found in subgingival plaque located at depths of more than 4 mm in periodontal pockets. T. denticola cells were found in the surface layers of subgingival plaque, and P. gingivalis were predominant beneath them. However, in the deeper subgingival plaque, the coexistence of P. gingivalis and T. denticola was observed. The present findings suggest that P. gingivalis and T. denticola play important roles in the pathogenicity of periodontal disease and provide the useful information for elucidating the pattern of colonization of microorganisms in the periodontal pocket. PMID- 7494176 TI - A feasibility study on the use of direct light silver stain compared with dark field microscopy for differential counting of subgingival plaque. AB - The purpose of the present study was to compare direct light microscopy using a silver stain, with dark field microscopy for differential counting of subgingival plaque samples from patients with periodontitis. The feasibility of using the staining method was assessed on subgingival plaque samples which were collected at 34 sites from patients with untreated adult periodontitis. Differential counts of plaque morphotypes assessed by both methods showed close agreement. The proportions of spirochaetes assessed by both methods were significantly associated with probing depth. The silver stain proved a simple, rapid and inexpensive method for differential counting of plaque composition. This method is suitable for possible use in general dental practice where smears could be made at the chair-side and allowed to dry ready for staining and counting at the dentist's convenience using a simple light microscope. A permanent record is also produced which could allow for comparison of samples from before and after treatment and could be used as an aid to patient motivation in suitable cases. PMID- 7494177 TI - Laminin and collagen IV distribution and ultrastructure of the basement membrane of the gingiva of the rat incisor. AB - The continuous growth of the rat incisor is associated with renovation of the junctional epithelium and resorption of the periodontal ligament. The circumdental papilla separates the connective tissue suffering resorption from the rest of the gingiva. Laminin and collagen IV were detected by the immunoperoxidase technique on the basement membrane of all regions of the gingival epithelium of the rat incisor, except the internal basal lamina and the internal surface of the circumdental papilla. The internal basal lamina is formed by a granular electron-dense material, without the organization of a typical basal lamina. Areas of the internal surface of the circumdental papilla, negative for laminin and collagen IV, lack the basal lamina. These data suggest that these molecules are not components of the dento-epithelial junction of the distal surface of the rat incisor. In addition, the basal lamina is absent or fragmented on the internal surface of the circumdental papilla, adjacent to the areas of the connective tissue undergoing resorption. PMID- 7494178 TI - Immunohistochemical study of linear gingival erythema from HIV-positive patients. AB - Severe forms of periodontal disease are frequent in patients with acquired immunodeficiency syndrome (AIDS). Linear gingival erythema (LGE) is a progressive disease described in HIV-positive patients and is considered to be an early stage of necrotizing periodontitis. Although clinical and microbiological differences are reported in LGE and non-specific gingivitis (NSG), a comparative immunopathological approach of both has not been performed yet. The purpose of this study was to compare relative populations of T-lymphocytes, B-lymphocytes, neutrophils, macrophages and IgG bearing plasma cells in gingival biopsies from sites exhibiting LGE and from sites exhibiting NSG. A biotin-streptavidin amplified system was used for identification of the following antigens: CD3 (T lymphocytes), CD20 (B-lymphocytes), elastase (neutrophils), CD68 (macrophages) and IgG (plasma cell's secretors of IgG). The results have demonstrated decrease proportions of T-lymphocytes, macrophages and high percentage of neutrophils and IgG bearing plasma cells in LGE. In contrast with NSG, many neutrophils cells in LGE were found inside oral gingival epithelium. Our results highlight the idea that progressive periodontal disease is not only characterized by increased tissue inflammation, but, in addition, by significant changes in the proportion of specific inflammatory cells. The high number of neutrophils along the gingival epithelium is probably associated with the severe gingival necrosis reported in AIDS patients. PMID- 7494179 TI - Specific cementum attachment protein enhances selectively the attachment and migration of periodontal cells to root surfaces. AB - A specific cementum attachment protein (CAP) was identified in human cementum and found to bind with high affinity to non-demineralized root surfaces, hydroxyapatite and fibronectin. Attempting to elucidate the biological function of this protein and its possible role in cementogenesis the capacity of CAP to promote selective cell migration towards and attachment of various periodontal derived cell populations to root surfaces in vitro was assessed. Human gingival fibroblasts (HGF), periodontal ligament cells (HPC), and alveolar bone cells (HABC) were labeled with [3H]Thymidine during their exponential growth phase. Root slices, 300 microns thick, were incubated with increasing concentrations of CAP. Untreated and fibronectin (FN) treated root slices served as negative and positive controls, respectively. Migration was assessed by placing root slices on confluent layers of labeled cells maintained in serum free medium and determining the number of cells migrated onto the root surface 3 days thereafter. Attachment was assessed by incubating root slices with labeled cell suspensions for 2 h and determining the number of attached cells. CAP promoted both cell migration and attachment dose dependently. HABC responded better than HPC and HGF to CAP treated root slices, and HPC response was higher than that of HGF. Cell attachment was dose dependently inhibited by synthetic RGD peptides. FN did not affect the migration of HGF, barely enhanced that of HABC, and was less potent than CAP at enhancing the migration of HPC. FN was more effective than CAP in promoting the attachment of HGF to root slices, but it was as potent as CAP in supporting the attachment of HPC and HABC.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7494180 TI - Influence of periodontal bacteria and disease status on V beta expression in T cells. AB - Some bacterial antigens such as S. aureus enterotoxins can selectively stimulate T cells that express specific V beta genes of the T cell antigen receptor (TCR). The purpose of this study was to investigate whether or not periodontal bacteria could similarly alter the expression of V beta families within the TCR complex. Peripheral blood mononuclear cells (PBMNCs) were isolated from 12 patients with early onset periodontitis and 11 periodontally-healthy controls. PBMNCs were incubated in media alone, or co-cultured for 48 h with heat-inactivated A. actinomycetemcomitans, P. gingivalis and P. intermedia. Expression of five V beta families (V alpha beta 2, V beta 5, V beta 6, V beta 8, and V beta 12) was determined by use of monoclonal antibodies. Mean unstimulated expression of V alpha beta 2 and V beta 8 was significantly higher (p < 0.05) in patients than healthy controls. Co-culture with the three bacteria resulted in significant changes (increases or decreases) in V beta expression in 27% of the trials. There were no significant differences in the number or direction of changes in samples from patients and controls. When compared to unstimulated controls, 18 significant increases but no decreases in the percentage of cell expressing V alpha beta 2, V beta 5 or V beta 6 were noted following co-culture with P. intermedia. Overall, co-culture with P. intermedia significantly (p < 0.05) up regulated expression of the five V beta families studied. These data suggest that periodontal bacteria may alter V beta expression within the T cell receptor complex. PMID- 7494181 TI - Towards gene therapy for cystic fibrosis. PMID- 7494182 TI - An investigation into the effects of preparation conditions and storage on the rate of drug release from pharmaceutical glyceride bases. AB - The effects of preparation conditions on the release of theophylline from Gelucires 50/13 and 55/18 have been investigated. Samples were prepared by melting physical mixes under controlled conditions, followed by either slow or fast (ambient) cooling to the solid state. A rapid cooling rate was shown to result in a slower release rate for 2% w/w dispersions in Gelucire 55/18, with the slow- and fast-cooled systems resulting in drug release via a diffusion and a mixed diffusion/erosion mechanism, respectively. At higher drug loading (30% w/w), the cooling rate did not effect the release characteristics. The erosion of Gelucire 50/13 was found to be more rapid for slowly cooled samples. Viscosity measurements were used as a means of assessing the chemical stability of the Gelucires, with evidence being found for degradation of Gelucire 55/18 on storage following heat treatment, while Gelucire 50/13 appeared to be stable. The effects of storage on the dissolution profiles of ambiently cooled systems were studied, with drug release from both bases increasing on ageing for up to 180 days. PMID- 7494183 TI - In-vitro and in-vivo studies of the diclofenac sodium controlled-release matrix tablets. AB - Controlled release matrix tablets for diclofenac sodium were developed in this study. Five matrix-tablet formulations were prepared by granulating two viscosity grades of HPMC (hydroxylpropylmethylcellulose) in varying ratios with water in the planetary mixer. The in-vitro dissolution tests indicate that all five matrix formulations prolong the release of diclofenac sodium. The main factors controlling drug release were the HPMC viscosity grade and the amount of HPMC used. The larger the amount of high viscosity grade HPMC used, the slower the resultant release rate of diclofenac sodium. There was no significant degradation of diclofenac sodium or change in drug release rate in any of the five formulations during a three-month period of stability testing. The sustained release ability of four formulations was further demonstrated in an in-vivo study in six healthy subjects. There were in-vitro/in-vivo correlations between Cmax, AUC0-14, and the time for 50 or 80% drug to be released. PMID- 7494184 TI - Studies on the in-vitro percutaneous penetration of indomethacin from gel systems in hairless mice. AB - The influence of co-solvents on the in-vitro percutaneous penetration of indomethacin from gel systems was studied using a simplex lattice experimental design. Gel formulations were prepared by gelling the vehicle mixture of water, either alcohol or isopropanol and either propylene glycol or PEG 400 with 1% w/w Carbomer 940. Hairless mouse skin was employed as the barrier in a Franz-type diffusion cell. The penetration rates at steady state for seven formulations were fitted to a polynomial equation based on this simple lattice method and a three dimensional plot was constructed. The formulation having the maximal penetration rate was determined to be the vehicle with a solvent ratio of water: alcohol: propylene glycol equal to 15:33:52, and which possessed a solubility parameter of 15 and a drug solubility of around 10 mg mL-1. When the solubility parameter of the vehicle was > 15, the drug solubility increased. However, the penetration rate decreased with an increasing solubility parameter. For those vehicles with a solubility parameter < 15, both the drug solubility and the penetration rate decreased with a decrease in the solubility parameter. There was shown to be an approximately 20-fold increase in the relative enhancement factor when using both alcohol and isopropanol, but only a threefold increase for both propylene glycol and PEG 400, when compared with water. PMID- 7494185 TI - Effects of ageing on the oral absorption of D-xylose in rats. AB - The effects of ageing on the oral absorption of D-xylose were investigated in rats. The pharmacokinetic analysis of D-xylose concentration in plasma after oral administration showed that the fraction absorbed was increased to 0.998 +/- 0.002 and 0.950 +/- 0.049, respectively, in old (52 weeks) and very old (102 weeks) rats, compared with 0.768 +/- 0.052 in young (9 weeks) rats, while the absorption rate constant was not significantly changed: 0.944 +/- 0.233, 0.844 +/- 0.143 and 0.725 +/- 0.004 h-1, respectively, in young, old and very old rats. The absorbed fractions estimated from faecal and urinary excretion were in agreement with those by the pharmacokinetic analysis. Thus, the present study demonstrated an increase in the extent of the oral absorption of D-xylose with ageing. The increase in the extent of absorption might be caused by a delay in the intestinal transit, because the absorption rate constant was unchanged. These results suggest potential increases with ageing in the fractions absorbed of hydrophilic drugs such as D-xylose where oral absorption is incomplete. PMID- 7494186 TI - Direct drug transport from the rat nasal cavity to the cerebrospinal fluid: the relation to the molecular weight of drugs. AB - To clarify the relationship between the direct transport from the rat nasal cavity to the cerebrospinal fluid (CSF) and the molecular weight of the drug, the transport of fluorescein isothiocyanate-labelled dextran (FD) with various molecular weights was investigated. FDs (average molecular weights 4,400 (FD4); 9,400 (FD10); 18,900 (FD20); 40,500 Da (FD40)) were administered nasally or intravenously to rats, and the concentrations in the plasma and the CSF were measured and compared. None of the FDs were detected in the CSF after intravenous administration. However, FD4, FD10 and FD20 were observed to appear in the CSF after nasal administration, whereas the concentration in the plasma was much lower than that after intravenous administration. FD40 was not detected even after nasal administration. In addition, the concentration of these FDs in the CSF decreased with the increase in the molecular weight of FDs. These findings show that drugs with a molecular weight up to at least 20,000 Da can be directly transported from the nasal cavity to the CSF and that the transport of FDs to the CSF is dependent on their molecular weights. PMID- 7494187 TI - PEG-coated poly(lactic acid) nanoparticles for the delivery of hexadecafluoro zinc phthalocyanine to EMT-6 mouse mammary tumours. AB - Hexadecafluoro zinc phthalocyanine (ZnPcF16), a second generation sensitizer for the photodynamic therapy of cancer, was incorporated in three vehicles: poly(D,L lactic acid) (PLA) nanoparticles, polyethylene glycol (PEG)-coated nanoparticles and a Cremophor EL (CRM) oil-water emulsion. Nanoparticles were prepared by the salting-out procedure. Biodistribution of the dye was assessed by fluorescence in EMT-6 mammary tumour bearing mice after intravenous injection of 1 mumol kg-1 ZnPcF16. Plain nanoparticles were rapidly retained by the reticuloendothelial system (RES) as reflected by the low area under the blood concentration-time curve (AUC0-168, 57 micrograms h g-1). Little tumour uptake of the dye was observed with this formulation. In contrast, PEG-coated nanoparticles displayed a reduced RES uptake, leading to significantly higher blood levels over an extended period (t1/2 30 h; AUC 0-168 227 micrograms h g-1) and enhanced tumour uptake. At 48 h post injection, tumour to skin and tumour to muscle concentration ratios reached 3.5 and 10.8, respectively. Blood levels of ZnPcF16 after administration as a CRM emulsion decreased faster than with PEG-coated nanoparticles (t1/2 12 h), but since no early liver uptake was observed, the AUC0-168 and the tumour uptake were only slightly lower. However, with the CRM formulation, a late liver uptake was observed, reaching 51% of the injected dose after 7 days. PMID- 7494188 TI - Contributions of beta-adrenoceptor subtypes to responses to isoprenaline in rat isolated distal colon. AB - The effects of the non-selective beta-adrenoceptor antagonist propranolol and the beta 1- and beta 2-adrenoceptor-selective antagonists, respectively CGP 20712A (((+/-)-[2-(3-carbamoyl-4-hydroxyphenoxy)-ethylamino]-3-[4-(1-meth yl-4- trifluoromethyl-2-imidazolyl)-phenoxy]-2-propanol hydrochloride)) and ICI 118,551 ((erythro(+/-)-1-(7-methylindan-4-yloxy)-3-isopropylamino butan-2-ol- hydrochloride)), on isoprenaline-induced inhibition of methacholine contractions in rat distal colon were investigated to determine the contributions of beta adrenoceptor subtypes to relaxation of smooth muscle. Longitudinal segments of rat distal colon were suspended in Krebs solution at 37 degrees C for isometric recording. The Krebs solution contained EDTA (30 microM), ascorbic acid (30 microM) and prazosin (0.1 microM) and was gassed with 95% O2-5% CO2. Isoprenaline produced a concentration-dependent inhibition of methacholine-induced contractions. Propranolol produced a small (4-6-fold) significant shift of the isoprenaline concentration-response curve at 0.003-0.01 microM. Larger shifts were produced by 0.3 microM (13-fold) and 1 microM (20-fold). CGP 20712A produced a small (3-5-fold) significant shift at 0.03-1 microM. ICI 118,551 produced small non-significant shifts (2-3-fold) at 0.03-1 microM. A combination of ICI 118,551 (0.3 microM) and CGP 20712A (0.1 microM) produced a 13-fold shift, a significantly greater shift than expected from the individual shifts. The shift produced by the combination of antagonists was slightly less than that produced by 1 microM propranolol (20-fold).(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7494189 TI - Acute moderate hypoxia in conscious rabbits: effect on hepatic cytochrome P450 and on reactive oxygen species. AB - This study aimed to document whether acute moderate hypoxia modifies the amount and activity of hepatic cytochrome P450 and in addition, induces changes in the production or the ability to neutralize oxygen reactive species (ORS). Rabbits were exposed to a low partial pressure of oxygen (12%) for 8 or 24 h, killed, and the amount and activity of cytochrome P450, lipid peroxidation, microsomal chemiluminescence and enzymatic scavenger activity were assessed in the liver. After 8 h of hypoxia, total amount but not the activity of cytochrome P450 was decreased, although after 24 h of hypoxia, both the amount and the activity of cytochrome P450 were decreased. Hypoxia for 8 h increased the activity of glutathione peroxidase. However, after 24 h of hypoxia, lipid peroxidation, microsomal chemiluminescence and superoxide dismutase activity were increased, while hepatic glutathione and glutathione peroxidase activity were reduced, modifications that suggest an enhanced presence of ORS. In in-vitro studies, an ORS generating system reduced the activity of cytochrome P450 and enhanced lipid peroxidation of hepatic microsomal membranes, supporting the view that ORS can impair cytochrome P450. The results of the present study show that hypoxia induces changes in the amount and activity of cytochrome P450, as well as in the production or the ability to neutralize ORS, and that these changes are time dependent. PMID- 7494191 TI - The effects of Danshen (Salvia miltiorrhiza) on warfarin pharmacodynamics and pharmacokinetics of warfarin enantiomers in rats. AB - The effects of Danshen (Salvia miltiorrhiza), a popular traditional Chinese medicinal herb on the pharmacokinetics and pharmacodynamics of R- and S-warfarin stereoisomers were studied in rats. After a single oral dose of racemic warfarin (2 mg kg-1), treatment with oral Danshen extract (5 g kg-1, twice daily) for 3 days significantly altered the overall pharmacokinetics of both R- and S-warfarin and increased the plasma concentrations of both enantiomers over a period of 24 h and the prothrombin time over 2 days. At steady-state levels of racemic warfarin (0.2 mg kg-1 day-1 for 5 days) the 3-day treatment of Danshen extract (5 g kg-1, twice daily) not only prolonged the prothrombin time but also increased the steady-state plasma concentrations of R- and S-warfarin. The results indicate that Danshen extracts can increase the absorption rate constant, area under plasma concentration-time curves, maximum concentrations and elimination half lives, but decreases the clearances and apparent volume of distribution of both R and S-warfarin. The pharmacokinetic and pharmacodynamic interactions of warfarin during co-treatment with Danshen extract observed in this study indicate an explanation for the clinically observed incidents of exaggerated warfarin adverse effects when traditional Chinese medicinal herbs or herbal products such as Danshen and Danggui (observed in a previous study) were co-administered. PMID- 7494190 TI - Iloprost: intracellular Ca(2+)-dependent contractile effect on isolated smooth muscle cells from guinea-pig ileum. AB - Prostaglandin E2 (PGE2) and iloprost induced a concentration-dependent contraction of smooth muscle cells isolated from the circular layer of guinea-pig ileum. PGE2- and iloprost-induced contractions were inhibited by the selective EP1-receptor antagonist, SC19220 (1-acetyl-2-(8-chloro-10, 11-dihydrodibenz (b,f) (1,4) oxazepine-10-carbonyl)-hydrazine), indicating the involvement of the EP1 subtype of the PGE2 receptor. When cells were incubated for 10 min in the presence of strontium (4 mM L-1), an inhibitor of the release of Ca2+ from intracellular store, the contractile effect of PGE2 and iloprost was inhibited. In contrast, incubation of cells in Ca/2+)-free medium, Ca(2+)-free medium plus EGTA, or in the presence of nifedipine, an organic Ca(2+)-channel blocker, did not alter the PGE2- and iloprost-induced contraction. These observations suggest that the myogenic effect of PGE2 and iloprost on intestinal smooth muscle is dependent on the release of intracellular calcium. PMID- 7494192 TI - Angiostasis and vascular regression in chronic granulomatous inflammation induced by diclofenac in combination with hyaluronan in mice. AB - Angiostasis and vascular regression in chronic granulomatous inflammation was assessed in mice induced with diclofenac in combination with hyaluronan. The local injection of 0.1 mL HYAL EX-0001 (0.18% diclofenac in 2.5% hyaluronan) reduced granulomatous development after six days treatment from 150.4 +/- 13.8 (0.18 saline) to 117.1 +/- 17.8 mg (dry weight, n = 10), but not significantly when compared with 0.1 mL 2.5% hyaluronan alone or diclofenac in 0.1 mL saline. Diclofenac administered in saline had no significant effect when compared with saline control. The vascular density, expressed as carmine content per mg dry weight tissue, in those animals treated with HYAL EX-0001 was also significantly reduced to 5.27 +/- 0.55 micrograms mg-1 (P < 0.1, n = 10) when compared with saline control (7.99 +/- 1.0), hyaluronan alone (7.20 +/- 1.0), and diclofenac in saline. (7.36 +/- 1.28). A similar profile of activity was seen on topical application except that all treatments did not affect granulomatous tissue development. On therapeutic dosing of mice daily with HYAL EX-0001 from day 7 after induction of the granulomatous tissue, the granulomatous tissue development was dramatically reduced from 111.67 +/- 4.40 mg (n = 14 on day 7) to 60.23 +/- 7.22 (P < 0.001, n = 8 on day 14) and 54.98 +/- 7.88 (P < 0.001, n = 8 on day 21). HYAL EX-0001 after 14 days of application significantly reduced granulomatous tissue mass when compared with the hyaluronan-dosed control on day 21 (89.58 +/- 7.49, P = 0.01, n = 8).(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7494193 TI - Brain distribution characteristics of xanthine derivatives and relation to their locomotor activity in mice. AB - The relationship between the brain distribution and motor activity in mice of the xanthines, theophylline, enprofylline, 1-methyl-3-propylxanthine (MPX) and oxpentifylline was investigated. Their plasma protein binding and hydrophobicity were also examined. When these xanthines were administered orally, enprofylline and oxpentifylline had no effect on motor activity. While theophylline increased motor activity over 10 mg kg-1, MPX caused a decrease in such activity over 10 mg kg-1. The protein-binding behaviour varied among these xanthines and was closely related to their hydrophobicity, which is represented as a logarithmic partition coefficient (log PC). MPX had the highest hydrophobicity, while oxpentifylline had the lowest. Brain distribution characteristics varied among these xanthines, with the rank order of their brain penetration ratio, calculated as the ratio of brain to unbound plasma concentrations, being theophylline > oxpentifylline > MPX > enprofylline. The inhibition constants (Ki) for adenosine A1 receptors and cyclic 3',5'-adenosine monophosphate (cAMP)-phosphodiesterase (PDE) of these xanthines were 44.6 and 134, > 1000 and 112, 26.4 and 49, and > 1000 and 111 microM for theophylline, enprofylline, MPX, and oxpentifylline, respectively. These findings suggest that the lack of effects of enprofylline and oxpentifylline on motor activity is probably due to their low brain penetration ratio or low adenosine A1 affinity in comparison with theophylline. The decrease in the motor activity by MPX may be, in part, mediated by cAMP or adenosine. PMID- 7494194 TI - Semicarbazide-sensitive amine oxidase activity in white adipose tissue of the insulin-deficient rat. AB - We have investigated whether the effects in white adipose tissue due to insulin deficiency might also be related to an alteration of histamine levels which are regulated by semicarbazide-sensitive amine oxidase. The lack of circulating insulin induced by streptozotocin produced, in rat white adipose tissue, a loss of affinity of the semicarbazide-sensitive amine oxidase for histamine oxidation. In parallel, a decrease of cell transformation, measured by glycerol-3-phosphate dehydrogenase activity and an augmented sensitivity to histamine lipolysis were observed. These findings could contribute to the understanding of histamine metabolism and function in diabetic rats and to the knowledge concerning amine oxidases in this animal pathology. PMID- 7494195 TI - Dexamethasone reduces the behavioural effects induced by baclofen in mice. AB - The present study examines the influence of dexamethasone on the behavioural effects induced by baclofen in mice. The behaviour elements considered were locomotor activity, motor co-ordination, catalepsy, stereotyped behaviour and antinociception. Baclofen (1.0-4.0-6.0 mg kg-1, i.p.) induced a significant reduction of all behavioural elements studied and an antinociceptive effect was recorded. Dexamethasone alone (0.1-0.5-1.0 mg kg-1, i.p.) did not induce significant changes in the behaviour elements considered. On the other hand, when the steroid was injected immediately before baclofen a significant reduction of baclofen's behavioural effects was found. Our results suggest a possible link between glucocorticoid and the GABA-ergic system. PMID- 7494196 TI - Acid-base properties of terbutaline in terms of protonation macro- and microconstants. AB - The acid/base chemistry of terbutaline was characterized at the molecular level in terms of protonation macroconstants and microconstants. The macroconstants were measured by potentiometry and calculated by standard evaluation methods. The stepwise macroconstant values were log K1 = 11.01, log K2 = 9.89, and log K3 = 8.57 at 25.0 degrees C and 0.2 M ionic strength. The microconstants were deduced using the relationships between macro- and microconstants and an appropriate data set of model compounds (resorcinol and phenylephrine). The molecule of terbutaline contains three ionizable functional groups. In the unprotonated form of the molecule, the two identical phenolate groups are slightly more basic than the secondary amino group, whereas the amino basicity significantly exceeds that of the phenolate site, when the other phenol is protonated. This is due to the large phenolate-phenolate intramolecular interaction. The phenolate-phenolate and the phenolate-amino interactivity parameters were found to be -1.21 and -0.41 log E units, respectively. PMID- 7494197 TI - Spectroscopic analysis of the binding of amylobarbitone, secbutobarbitone, pentobarbitone, phenobarbitone and quinalbarbitone to human serum albumin. AB - The binding of five barbiturates: amylobarbitone, secbutobarbitone, pentobarbitone, phenobarbitone and quinalbarbitone to human serum albumin (HSA) was studied by difference spectroscopy and spectrofluorimetric titrations. There were no changes in the HSA spectral properties. Our result suggest that there are two classes of binding sites on HSA for these barbiturates. A detailed investigation of the effect of their binding to HSA by deconvoluted spectra, suggests that the interaction of barbiturate-HSA takes place principally on the subdomain IIIA of HSA. PMID- 7494199 TI - Where are we now and where are we headed? PMID- 7494198 TI - Acid-base equilibria of 1,4-benzodiazepine 4-oxides by spectrophotometry. AB - Chlordiazepoxide (a 1,4-benzodiazepine 4-oxide) is an anxiolytic/hypnotic drug in clinical use. It was reported to be predominantly protonated at the N-oxide oxygen in acidic aqueous solutions at pH << 4.6 (pKa). We have studied the acid base equilibria of three 1,4-benzodiazepine 4-oxides (chlordiazepoxide, diazepam 4-oxide, and nordiazepam 4-oxide) by ultraviolet-visible spectrophotometry. The results indicate that chlordiazepoxide is not protonated at the N-oxide oxygen, but rather at the nitrogen of an imine bond between C2 carbon and its nitrogen substituent in acidic media. PMID- 7494201 TI - Licensed practical nursing education transformation in progress. PMID- 7494200 TI - The monster has a name. PMID- 7494202 TI - LPNs and delegating. PMID- 7494203 TI - Legal implications of dual licensure: a study of North Dakota. PMID- 7494204 TI - Building a nursing career. PMID- 7494205 TI - Are unlicensed assistive personnel a threat to staff and patients? PMID- 7494206 TI - Pleasant dreams at last. PMID- 7494207 TI - 5th International Evoked Potentials Symposium. Milan, Italy, 4-7 September 1994. Proceedings. PMID- 7494209 TI - Non-invasive multielectrode array for high resolution sampling of scalp-recorded potential fields. AB - A new non-invasive electrode array has been devised to record electrical signals on the scalp. The electrodes are arranged every 10 mm in a 4 x 8 grid, thus totalling 32. Electrical coupling is assured by a column of conductor gel with a diameter of 2 mm. The electrode array has been used to record multichannel scalp potentials evoked by stimulation of the median nerve at the wrist. It was positioned on the parietal region with the midline corresponding to the external acoustic meatus. The high spatial resolution of this device allows a detailed spatial sampling of the scalp potential field where the electrical activity arising from the parietal and premotor cortex is predominant. Scalp potentials with the system are compared with those with the 10/20 system. The improved spatial resolution allows more precise localization of minimum and maximum of the potential field, though the overall distortion introduced by aliasing in the 10/20 system is not striking. Instead, the amplitudes of the fields are considerably affected by insufficient spatial sampling. PMID- 7494208 TI - Use of DC recording in the demonstration of functional specialization. AB - This paper describes a rationale and methodology for the use of DC recording techniques to reveal patterns of cerebral functional specialization. Results from a number of recent experiments are described. In each of these studies 15 channels of DC potential were recorded from a widely spaced array of scalp mounted Ag/AgCl electrodes. The duration of single trial recording was always 40 s although the paradigms involved continuous performance of a complex cognitive task. In this series of experiments we used the following tasks: both visually and aurally presented word target detection with classification on a semantic basis, recognition of 'famous' faces and discrimination of animal versus non animal sounds. In each of these tasks the rate of presentation has been systematically varied to evaluate quantitative aspects of task demand. In general, the results from these studies demonstrate that DC recording does provide a method for assessing the degree of activation of different cortical regions at the same time. However, it appears that additional transformation of the data may be required to separate out a number of overlapping slow potentials to increase the sensitivity to more subtle aspects of the cognitive demand. Two different methods to perform the separation are described and applied. PMID- 7494210 TI - Measurement of the activating function of magnetic stimulation using combined electrical and magnetic stimuli. AB - The technique of combined magnetic and electrical stimulation of a peripheral nerve was used to determine the effectiveness of the combined stimulus and its dependence on the relative positioning of electrodes and stimulator coil along the axis of the nerve. The results were used to determine the magnetic stimulation-activating function of a long, straight nerve in the arm, and are shown to be consistent with published theoretical models constructed under conditions of simplified tissue geometry. With appropriate positioning of the two stimulators and similar tissue current waveforms, both enhancement and inhibition of an electrical stimulus were demonstrated and the maximum amplitude of the combined stimulus approached the arithmetic sum of that produced by each stimulator individually. PMID- 7494211 TI - A new procedure for the registration of the visual-evoked cortical potential by multichannel recording of the gradient distribution. AB - A new bipolar multichannel visual-evoked cortical potential (VECP) procedure is presented, which can supply objective information on visual field losses. The electrical potential is recorded from 11 electrodes applied to the back of the head in the form of an equidistant rectangular grid. Each neighbouring pair of electrodes feeds one of 14 bipolar channels. The adjacent horizontal and vertical channels are used to calculate an approximation to the direction and amplitude of the electrical field gradients. The gradient distribution is represented by a map of arrows (gradient map) for every instant of the sweep, so that the whole sweep can be plotted as a time series of gradient maps. The maps are easy to scale and are well suited for visual evaluation. Twenty normal subjects were stimulated using checkerboard reversal, partial field patterns to simulate visual field defects. The stimulated area varied between full-, half- and quarter-field, and the particular area stimulated could be clearly seen in the resulting gradient maps. Additionally, we developed a computerized classification procedure that detected 86% of the disturbed visual fields from the gradient recordings. PMID- 7494212 TI - Event-related potential map differences depend on the prestimulus microstates. AB - The dependency of the landscapes of visually evoked, 47-channel, event-related potential (ERPs) on the functional microstates (momentary map landscape) just before stimulus arrival was investigated, in 12 volunteers. The prestimulus microstates were determined using the map at the last peak of Global Field Power before the stimulus. The landscapes of these maps were described by the electrode locations of the positive and negative extreme potentials, and assigned to basic classes. The two most frequently occurring map-classes were used (left anterior right posterior, and right anterior-left posterior). ERP map series were averaged for each subject and each prestimulus microstate class. The Randomization-Monte Carlo MANOVA test was used to test the significance of the difference between the ERP map landscapes at each sample point (n = 128, 500 ms) associated with the two prestimulus microstates. At 16 samples the difference was significant at p < 0.05. The longest uninterrupted sequence (n = 9) of significant differences occurred between 164 and 195 ms, i.e. during the conventional component P200. The results demonstrate that the brain electric microstate at stimulus arrival crucially influences the active neuronal populations that contribute to the ERP. This suggests that the processing of information will differ as a function of the momentary brain microstate at information arrival. PMID- 7494213 TI - Temporal segmentation and multiple-source analysis of short-latency median nerve SEPs. AB - Short-latency (10-50 ms) median nerve somatosensory evoked potentials (SEPs) from four normal subjects were analysed by means of temporal segmentation techniques and source derivation methods. In each case the responses were recorded using 32 electrodes. Dipolar optimization was carried out with a time-varying technique, using three different approaches: regional source estimation, spherical source estimation (one radial and one tangential component), and multiple dipolar approach. This was to assess the relative influence on the dipolar solution of the different optimization techniques. The effect of the different number of channels in the estimation procedures has been also investigated. The methods of optimization are crucial, particularly for the orientation of P22. In all cases the source location estimated with the 32-electrode montage was shifted towards the centre of the spheres. PMID- 7494214 TI - Monitoring evoked potentials during surgery to assess the level of anaesthesia. AB - The hypnotic and analgesic components of anaesthesia can be assessed using middle latency auditory evoked potentials (MLAEPs) and somatosensory evoked potentials (SEPs). To monitor these potentials reliably during clinical anaesthesia, we have developed an evoked potential (EP) system based around a portable personal computer, a DSP board and an isolated pre-amplifier unit. Unlike many currently available systems, this amplifier is largely immune to diathermy interference due to excellent isolation via a digital fibre optical link, small size and RF screening and filtering. The pre-amplifier unit has integral auditory and somatosensory stimulators, and automatic calibration and impedance checking. Stimulus intensity and profile are under software control and SEP stimulus level is constantly monitored. The unit is powered by two AA cells and battery status continuously monitored by the PC software. Up to eight channels of EEG may be recorded and displayed in a smoothly scrolling window and as moving average MLAEPs and SEPs. PMID- 7494215 TI - Automatic perimetry and visual P300: differences between upper and lower visual fields stimulation in healthy subjects. AB - Human visual-evoked potentials (VEPs) from upper and lower hemifield stimulation are thought to reflect the anatomical and functional differences between the hemiretinas and corresponding visual pathways. Conflicting results have, however, been reported in topographic studies on the putative cortical generators. We have estimated by automatic perimetry (Octopus 2000R, Program 32) and compared the sensitivity thresholds of lower and upper hemifields of the retina in 12 healthy subjects with no history or evidence of visual or neurological diseases. A visual P3 that is linked to cognitive function was recorded in an odd-ball paradigm with presentation of high-contrast checkerboards at two different spatial frequencies at 20 degrees eccentricity in each hemifield. VEP and P3 were recorded at O1 and O2 and at Cz according to the 10/20 international system. Lower sensitivity thresholds were found, and higher VEP and event-related potential (ERP) amplitude values were obtained when stimulating the lower, compared with the upper, visual hemifield. The results are consistent with previous findings and anatomical and physiological evidence in animals and man. Interactions between perceptive process in the visual system and higher cognitive functions are a possible explanation for this finding. PMID- 7494216 TI - Validation of a detailed computer model for the electric fields in the brain. AB - A computer model has been designed for the calculation of the electrical fields in the head, based on the finite difference method. This method has not previously been applied for head modelling. The model was validated by using three concentric spheres and comparing it with an analytic model. Three levels of accuracy were tested. The forward solutions show that the finite difference algorithm works correctly and, by selecting the size of the volume elements properly, accurate results are obtained. The model will be applied to accurate and realistic geometries of the human head obtained from magnetic resonance images. PMID- 7494217 TI - A multichannel, model-free method for estimation of event-related potential amplitudes and its comparison with dipole source localization. AB - We present a multichannel, model-free method for estimation of event-related potential (ERP) amplitude ratios and amplitudes using singular value decomposition (SVD), and compare with the Dipole Components Model (DCM). When the ERPs are generated by a single or multiple dipoles with equal amplitude ratios, the SVD method is superior to DCM in terms of reliable estimation of amplitude and is comparable with DCM for reliable and unbiased estimation of amplitude ratios. We show that dipole model misspecification leads to unbiased amplitude ratios and biased amplitudes when the ERP data sets are (1) generated and fit with a single dipole, or (2) generated by N dipoles with equal amplitude ratios and fit with M < or = N dipoles, because the effect of model misspecification 'cancels' for a ratio. Similarly proof that DCM estimates amplitude ratios more reliably than amplitudes for these cases is given. PMID- 7494218 TI - Geographic patterns of breast cancer among American women. PMID- 7494219 TI - Mutant Ras proteins and peptides: bad news for tumors. PMID- 7494221 TI - Use of organ-sparing treatments varies internationally. PMID- 7494220 TI - Additional evidence implicating moist snuff as a potent carcinogen. PMID- 7494223 TI - NSABP halts B-14 trial: no benefit seen beyond 5 years of tamoxifen use. PMID- 7494222 TI - Survey shows M.D.s' ambivalence about maximum androgen blockade. PMID- 7494224 TI - New screening tool maps genetic patterns of solid cancers. PMID- 7494225 TI - EU cancer code tuned-up, targets GPs and the public. PMID- 7494226 TI - Survivors' congress promotes cancer research. PMID- 7494228 TI - Geographic variation in mortality from breast cancer among white women in the United States. AB - BACKGROUND: For several decades, mortality from breast cancer has been higher in the northeastern part of the United States than in other regions, particularly the South. Rates have also been somewhat higher in the Midwest and West than in the South, especially among older women. The reasons for these geographic variations are not well understood. PURPOSE: The objective of this study was to evaluate geographic differences in U.S. breast cancer mortality rates in 1987, after taking into account regional differences in the distribution of recognized breast cancer risk factors (e.g., late age at first live birth) and certain prognostic factors (e.g., mammography use). METHODS: The 1987 breast cancer mortality rates for four regions of the country were obtained from the National Center for Health Statistics. Regional data on the distribution of breast cancer risk factors were obtained from 1987 National Health Interview Cancer Epidemiology Supplement interviews with 9778 white women aged 20-79 years. Regional data on the distribution of mammography use were obtained from 1987 National Health Interview Cancer Control Supplement interviews with 3795 white women aged 50-79 years. RESULTS: Age-adjusted mortality ratios (MRs) among women 50 years and older were 1.15, 1.18, and 1.30 in the West, Midwest, and Northeast, respectively, compared with the South. Corresponding MRs among women 20-49 years old were 1.01, 1.08, and 1.07 in the West, Midwest, and Northeast, respectively, compared with the South. After adjustment for recognized risk factors and certain prognostic factors, MRs among older women were 1.13 (95% confidence interval [CI] = 1.04-1.23), 1.08 (95% CI = 1.01-1.16), and 1.13 (95% CI = 1.04-1.23) in the West, Midwest, and Northeast, respectively, compared with the South. Corresponding MRs among younger women were 0.94 (95% CI = 0.76-1.16), 1.05 (95% CI = 0.92-1.18), and 0.99 (95% CI = 0.86-1.14), respectively. CONCLUSION: Before adjustment for regional differences in recognized risk factors and prognostic factors, mortality excesses among younger women in the Northeast, Midwest, and West were less than 10% compared with the South. After adjustment, MRs were near unity for all regions. Among older women, the excess mortality was more substantial before adjustment for relevant factors, ranging from 15% in the West to 30% in the Northeast. Approximately 50% of the excesses in the Northeast and Midwest and 10% of the excess in the West could be explained on the basis of regional differences in the prevalence of recognized breast cancer risk factors and prognostic factors. After adjustment for these factors, the magnitude of excess in breast cancer mortality in the Northeast (13%) was comparable to that in the West (13%) but still slightly higher than that in the Midwest (8%). PMID- 7494227 TI - Meeting highlights: a reappraisal of research results for the local treatment of early stage breast cancer. PMID- 7494229 TI - Induction of T-cell immunity against Ras oncoproteins by soluble protein or Ras expressing Escherichia coli. AB - BACKGROUND: Point mutations in the ras proto-oncogene that activate its oncogenic potential occur in approximately 30% of human cancers. Previous studies have demonstrated that T-cell immunity against some forms of mutant Ras proteins could be elicited, and some effectiveness against tumors expressing activated Ras has been reported. PURPOSE: The goal of this study was to determine if immunization of mice with two forms of mutant Ras protein can induce high levels of Ras mutation-specific T-cell immunity in vitro and tumor regression in vivo. METHODS: Mice (BALB/c or C3H/HeJ) were immunized subcutaneously at 2-week intervals with purified Ras oncoproteins mixed with the immunologic adjuvants Antigen Formulation or QS-21, both of which have been shown to enhance the induction of T cell-mediated immunity when included as components of soluble protein vaccines. In some experiments, mice were immunized directly with heat-killed Escherichia coli that had been induced to express one of the mutant Ras proteins. Spleen cells plus lymph node cells from Ras-immunized mice were tested in vitro for lysis of syngeneic Ras-expressing tumor cells and proliferation in response to mutant Ras peptides. For some of the cytolytic activity experiments, the spleen cells were grown under TH1 conditions (growth in presence of interleukin 2, interferon gamma, and an antibody directed against interleukin 4 to stimulate a cell-mediated immune response) or TH2 conditions (growth in presence of interleukins 2 and 4 to stimulate a humoral immune response). The specificity of immunity was examined in vivo by challenge of Ras-immunized mice with syngeneic tumor cells expressing mutant Ras oncoproteins (HaBalb, i.e., BALB/c mouse cells expressing Ras with arginine substituted at amino acid position 12 [Arg 12 Ras]; C3HL61, i.e., C3H/HeJ mouse cells expressing Ras with leucine substituted at position 61 [Leu 61 Ras]). Ten mice per group were used in each experiment. RESULTS: Proliferative and cytolytic T-cell responses directed against the Arg 12 Ras protein were generated in BALB/c mice, resulting in protection against challenge with cells expressing Arg 12 Ras and therapeutic benefit in mice bearing established tumors expressing this protein. In C3H/HeJ mice, high levels of cytolytic and proliferative responses were induced against Leu 61 Ras. Immunization with heat-killed E. coli genetically engineered to express Leu 61 Ras also led to the induction of anti-Ras T-cell immunity. T cells grown under TH1 conditions were cytolytic against Ras-transformed tumor cells, whereas those grown under TH2 conditions were not. CONCLUSIONS: Immunization as described here leads to Ras mutation-specific antitumor immunity in vitro and in vivo, with therapeutic efficacy in an established tumor model. PMID- 7494230 TI - Five leading U.S. commercial brands of moist snuff in 1994: assessment of carcinogenic N-nitrosamines. AB - BACKGROUND: Moist snuff is the only tobacco product in the United States with increasing sales (an increase of 38.4% between 1981 and 1993) and with increased consumption, primarily by male adolescents aged 12-18 years old and young adults aged 19 years old or older. It is known from previous studies that levels of nicotine and the proportion of unprotonated (free) nicotine, as well as the pH, which affects nicotine delivery, vary considerably among the leading snuff brands. Whether concentrations of major carcinogens, such as the nicotine-derived tobacco-specific N-nitrosamines (TSNAs), like N'-nitrosonornicotine (NNN) and 4 (methylnitrosamino)-1-(3-pyridyl)-1-butanone (NNK), also vary among these brands has not been determined previously. PURPOSE: Our purpose was threefold: 1) to determine the concentrations of major carcinogenic nicotine-derived N nitrosamines in each of the five most popular moist snuff brands; 2) to analyze the quantitative differences in the various snuff components (e.g., NNN) between two major brand categories: a category comprising the brands known to have high levels of unprotonated nicotine (Copenhagen, Skoal fine cut, and Kodiak) versus a category comprising the brands known to have low levels (Hawken and Skoal Bandits); and 3) to compare the differences in the concentrations of nicotine (previously determined), NNN, NNK, and total TSNAs between these two major brand categories. METHODS: Three boxes of each of the five leading U.S. moist snuff brands were bought in July 1994 from retailers in six areas and transferred immediately to the analytical laboratory. After extraction, N-nitrosamino acids and TSNAs were determined on a gas chromatograph interfaced with a thermal energy analyzer (GC-TEA) and integrator. Each 5-g sample of ground, freeze-dried tobacco was extracted twice, and each extract was analyzed twice by GC-TEA. All P values reported are two sided. RESULTS: Copenhagen, Skoal fine cut, and Kodiak as a group had statistically significant higher levels of nicotine (P = .0017), NNN (P < .0001), NNK (P = .0119), and total TSNAs (P < .0001) than the Hawken and Skoal Bandits group. Concentrations (means +/- SD) of nicotine, NNN, NNK, and total TSNAs comparing the two major brand categories are as follows: nicotine--11.6 +/- 1.01 mg/g versus 6.96 +/- 3.62 mg/g (P = .0017), NNN--7.74 +/- 1.70 micrograms/g versus 4.17 +/- 1.35 micrograms/g (P < .0001), NNK--1.23 +/- 0.68 micrograms/g versus 0.61 +/- 0.41 micrograms/g (P = .0119), and total TSNAs--14.3 +/- 3.82 micrograms/g versus 6.3 +/- 2.56 micrograms/g (P < .0001). CONCLUSIONS: The three leading U.S. snuff brands (Copenhagen, Skoal fine cut, and Kodiak; making up 92% of the U.S. market) showed not only high levels of pH, nicotine, and unprotonated (free) nicotine, but also high concentrations of the strongly carcinogenic TSNAs in comparison with the fourth and fifth best selling moist snuff brands, Hawken and Skoal Bandits (3% of the U.S. market). PMID- 7494231 TI - Immunohistochemical analysis of the p16INK4 cyclin-dependent kinase inhibitor in malignant mesothelioma. AB - BACKGROUND: The identification in 1994 of the CDKN2 gene as a target for mutations in a wide range of human cancers, including malignant mesothelioma, has been controversial because subsequent studies have detected a lower frequency of CDKN2 gene mutations in primary tumors than in cultured cell lines. These reports raised the hypothesis that another gene, distinct from CDKN2, might be the target of the chromosome 9p21 deletions frequently observed in these tumors. PURPOSE: To address whether inactivation of CDKN2 function is an essential event in the etiology of malignant mesothelioma, we examined p16INK4 protein expression in primary thoracic mesotheliomas, in nonmalignant pleural tissues, and in independent mesothelioma cell lines. We also studied the growth rate of tumor cell lines following stable transfection of CDKN2 gene. METHODS: Retinoblastoma (Rb) and p16INK4 protein expression was determined by immunohistochemical analysis from archival paraffin specimens of 12 primary thoracic mesotheliomas and a nonmalignant pleural biopsy specimen. In addition, protein immunoblot analysis for Rb and p16INK4 expression was conducted on 15 independent mesothelioma cell lines, and the ability of a transfected CDKN2 gene to suppress the growth of the mesothelioma cell lines H2373 and H2461 in vitro was examined. RESULTS: We demonstrated abnormal p16INK4 expression in 12 of 12 primary mesothelioma specimens and in 15 of 15 mesothelioma cell lines. All tumor specimens and the tumor cell lines showed expression of wild-type Rb protein. In addition, we have confirmed the ability of a transfected CDKN2 gene to suppress growth of two independent mesothelioma cell lines. CONCLUSIONS: Immunohistochemical analysis of the p16INK4 gene product is feasible in archival biopsy samples. With this analysis, CDKN2 gene inactivation can be determined in tumors that are contaminated with nonmalignant cells. Furthermore, since loss of p16INK4 protein expression can result from both genetic (gene mutations) and epigenetic (abnormal DNA hypermethylation) mechanisms, as we and others have shown recently, examination of protein expression is a highly sensitive method for analyzing the CDKN2 status in large numbers of tumor samples. IMPLICATIONS: This study suggests that inactivation of the CDKN2 gene is an essential step in the etiology of malignant mesotheliomas. Defining the role of the p16INK4:Rb tumor suppressor pathway and its immediate downstream substrates will be an important goal in designing future therapeutic strategies. PMID- 7494232 TI - Irinotecan (CPT-11) and characteristic mucosal changes in the mouse ileum and cecum. AB - BACKGROUND: Irinotecan--or CPT-11; 7-ethyl-10-[4-(1-piperidino)-1-piperidino] carbonyloxy-camptotheci n--is an inhibitor of DNA topoisomerase I and is clinically effective against several cancers. A major toxic effect of CPT-11 is severe diarrhea; however, the exact mechanism by which the drug induces diarrhea has not been established. Cisplatin (CDDP; cis-diamminedichloroplatinum) and CPT 11 exhibit synergistic antitumor activity and have been used in combination chemotherapy regimens. Single-agent chemotherapy with conventional doses of CDDP does not cause clinically relevant diarrhea. PURPOSE: To elucidate the mechanisms of induction of diarrhea by high-dose CPT-11 and to compare them with those of diarrhea induced by high-dose CDDP, we used histopathologic and immunohistochemical methods to examine the intestines of mice treated with either CPT-11, CDDP, or saline (control). METHODS: Male ICR mice were administered intraperitoneally either 100 mg/kg CPT-11 daily for 4 days, 10 mg/kg CDDP daily for 3 days, or phosphate-buffered saline (control) daily for 4 days (10 mice per group). Preliminary experiments indicated that diarrhea was induced in mice approximately 6 days after administration of CPT-11 or CDDP; therefore, in the experiments described, animals were killed 6 days after the first dose. Serial paraffin-embedded sections of the intestine were stained with hematoxylin-eosin, Grimelius (to identify endocrine cells), or high-iron diamine-alcian blue (stains sialomucin blue and sulfomucin brown-black). Immunohistochemical analyses were performed with the use of anti-proliferating cell nuclear antigen (anti-PCNA; to assay proliferation), anti-Le(y) (BM-1; indirect measure of apoptosis), and anti synaptophysin antibodies (to identify the enteric nervous system and enterochromaffin cells). A terminal deoxynucleotidyl transferase-mediated deoxyuridine triphosphate-biotin nick end-labeling (TUNEL) method was used to detect DNA fragmentation in situ (i.e., apoptosis). The concentrations of two intestinally active secretogogues, plasma serotonin and vasoactive intestinal polypeptide, were also measured. RESULTS: The levels of plasma intestinal hormones were similar in control, CPT-11, and CDDP groups. No active necrotic changes were observed in the intestines of CPT-11- and CDDP-treated mice, even though marked thinning of the intestinal walls was observed in both cases. The intestines of CPT-11-treated mice, but not those of control or CDDP-treated mice, were characterized by epithelial vacuolation of the ileum (associated with increased apoptosis as measured by BM-1 and TUNEL) and goblet-cell hyperplasia with excessive amount of sulfomucin in the cecum (suggesting induction of differentiation). By contrast, CDDP treatment of mice reduced the number of villi in the jejunum and destroyed crypt cells containing large Paneth (secretory) granules in the ileum. CONCLUSIONS: CPT-11 may produce characteristic mucosal changes in the intestine by inducing apoptosis and cell differentiation. The observed changes are likely to cause malabsorption of water and electrolytes and hypersecretion of mucin. These structural and functional effects are probably the main causes of CPT-11-induced diarrhea. CDDP appears to cause diarrhea in mice by causing diffuse mucosal damage in the intestines. PMID- 7494233 TI - Evaluating glioma therapies: modeling treatments and predicting outcomes. AB - BACKGROUND: Intra-arterial chemotherapy with carmustine (BCNU) and interstitial radiation therapy with the use of stereotactically placed 125I sources are aggressive local therapies for malignant glioma. These therapies emerged in the 1980s and both appeared promising in phase II studies but yielded disappointing results in subsequent randomized controlled trials by the Brain Tumor Cooperative Group (BTCG). Florell and colleagues had prepared us for the possibility that brachytherapy would have less impact on survival than anticipated from the phase II experience by demonstrating that patients who were judged eligible for interstitial radiation, but treated conventionally, lived significantly longer than those who were ineligible and had better than average outcomes. PURPOSE: To further examine the impact of patient selection on outcome, we used the database of Florell et al. to assess the survival of patients with malignant glioma who were eligible or ineligible for chemotherapy by three intra-arterial methods, one of which was similar to that employed by the BTCG in its randomized, controlled trial evaluating intra-arterial BCNU. METHODS: The medical records and computed tomography (CT) scans of 102 consecutive patients with malignant glioma receiving standard treatment (i.e., maximum feasible surgical resection, external-beam radiotherapy, and often adjuvant systemic chemotherapy) at a single cancer center in Canada during the calendar years 1988 and 1989 were used for this analysis. Based on CT imaging and blind to outcome, an interventional neuroradiologist decided which patients were eligible or ineligible for intra-arterial chemotherapy via injection of two major arteries, via injection of one major artery, or via selective middle-cerebral artery injection. A Karnofsky performance score of greater than or equal to 60 was required. The percent of eligible patients, the median survival time, and the distribution of prognostic factors were analyzed for each group of eligible and ineligible patients. Median survival times were compared with the use of the generalized Wilcoxon (Breslow) test. All P values were based on two-tailed tests. RESULTS: For two-vessel treatment, 72.5% of the patients (74 of 102) were eligible; the eligible patients on average lived longer than the ineligible patients (14.8 versus 3.5 months; P < .00001). For one-vessel treatment, 48% of the patients (49 of 102) were eligible; again, the eligible patients lived longer than the ineligible patients (18.4 versus 5.1 months; P < .00001). For middle-cerebral artery treatment, 30% of the patients (31 of 102) were eligible, and these eligible patients did live somewhat longer than the ineligible patients, but this result did not reach statistical significance (13.6 versus 9.9 months; P = .1304). Trends were similar for patients with glioblastoma multiforme and anaplastic glioma. The median duration of survival was 11.4 months for all patients. CONCLUSIONS: Patients who were eligible for intra-arterial chemotherapy lived significantly longer or somewhat longer (depending on the selection criteria used) than patients who were ineligible and had better than expected outcomes. Patients who were judged eligible for intra-arterial chemotherapy by the two-vessel method and the control group in the BTCG phase III trial of intra-arterial chemotherapy had similar median survival times (14.8 versus 14.0 months). IMPLICATIONS: Modeling treatments with the use of a comprehensive clinical and imaging database of unselected, conventionally treated patients may help investigators decide if new therapies warrant definitive evaluation in randomized trials by measuring the degree to which patient selection may have enhanced phase II study outcomes. PMID- 7494234 TI - Bacteriology of maxillary sinuses in rabbits. AB - To study whether normal sinus flora exist in normal sinus, we used 40 sinuses from 20 New Zealand white rabbits as the animal model. The histological examination was performed on sinus mucosa from each sinus and bacteria from these sinuses were cultured. Surprisingly, 70% of them (28 sinuses) were found to have sinusitis. These sinuses not only contained bacteria in culture but also showed various degree of histological inflammatory reaction in mucosa examination. Of the other 12 normal or near normal sinuses, only two showed positive bacterial culture, and ten of them showed negative bacterial culture either from discharge or mucosa studies. Accordingly, these data suggest that these two studies (bacterial culture and histological mucosa examination) are consistent. Thus, from this study it was concluded that there is no bacteria or few bacteria in normal sinus. PMID- 7494235 TI - An investigation of junior high students' attitudes toward amphetamine use and related forms of behavior. AB - The purpose of the present study was to investigate the attitudes toward amphetamine use and some forms of behavior related to it. Two different kinds of junior high schools from southern Taiwan participated in this study. Five hundred and sixty-nine students supplied urine samples and filled in questionnaires. The results indicated that the prevalence rate of amphetamine use was 4.0% and the subjects had a fair level of cognition and very negative beliefs toward amphetamine use. Discriminant analyses indicated that access to amphetamine and the unlikelihood of being caught when using amphetamine were the two most important and meaningful variables in the distinction of previous using experience. School, grades, opinions about amphetamine use, the ease with which amphetamine could be obtained, and the severity of misbehavior were the most important variables in the prediction of the probable intention to use it, if amphetamine were available. However, because of the limited number of amphetamine users found in the study, any conclusions based on the results should be used cautiously. Strategies of prevention are suggested. PMID- 7494236 TI - Prevalence and size of simple hepatic cysts in Taiwan: community- and hospital based sonographic surveys. AB - Simple hepatic cysts are now being diagnosed more frequently with the widespread availability of sonographic imaging. Among the general population, the prevalence was reported to be between 0.1 and 2.5%, more so in women, and more often in the right lobe. We conducted this large scale community-based sonographic screening on simple hepatic cysts to explore the age- and sex-specific prevalence in Taiwan. We also conducted another hospital-based study to record the size of simple hepatic cysts. A total of 3,600 subjects in 8 communities were recruited and 156 simple hepatic cysts in 132 study subjects were detected. The overall prevalence is 3.60%. An increasing prevalence with age is demonstrated, ranging from 0.83% from below the age of 40 up to 7.81% of patients over 60 years old. The sizes of 219 simple hepatic cysts of 167 patients out of the hospital-based 5,893 patients were recorded in detail; 53% of cysts were of a diameter of between 1 and 3 cm, and only 7% were larger than 5 cm. PMID- 7494238 TI - Application of ABO genotyping using polymerase chain reaction method to assess engraftment in bone marrow transplantation--a case report. AB - We described a case of acute promyelocytic leukemia with O blood type who received bone marrow transplantation during a second remission from his HLA identical, B blood type sibling. Using PCR genotyping of the ABO group and analysis of the variable number of tandem repeats of the D1S80 locus, we could quickly document the successful early engraftment in this patient. We emphasize that the use of PCR ABO genotyping is a quick and valuable method which could be applied to assess the engraftment and its follow up in those transplant cases with donors of mismatched blood groups. PMID- 7494237 TI - Superior vena cava syndrome in lung cancer: an analysis of 54 cases. AB - Traditionally, superior vena cava syndrome (SVCS) has been recognized as an oncologic emergency, and with clinical suspicion of the syndrome, tissue diagnosis was often delayed due to possible complications in diagnostic procedures and immediately threatening of life. Previously, local radiotherapy was regarded as the best immediate strategy for management of the condition. We have analyzed 54 lung cancer patients with SVCS in the past 6 years. Our results show that dyspnea (34 cases, 63%) and facial swelling (29 cases, 54%) are the two most common symptoms. The most frequent physical finding was venous distension of the neck (35 cases, 65%). The chest X-ray findings also showed a large ratio of superior mediastinal widening (26 cases, 48%). Fine needle aspiration of palpable lymph node (20 cases, 37%) and trans-thoracic needle aspiration guided by ultrasound (US) (8 cases, 14%) made diagnosis of at least half of the cases possible (28 cases, 51%). Both of these procedures are safer and easier than other invasive methods of examination. Of the 54 patients, small cell carcinoma constituted the majority of the cases (23 cases, 43%) and, with combination chemotherapy, there was a good response rate and a longer survival time (7.4 months) as compared to that of non-small cell carcinoma (3.7 months) treated by radiotherapy. We conclude that lung cancer with SVCS could be quickly and safely diagnosed by needle aspiration of the palpable lymph node or trans-thoracic needle aspiration guided by US, and that with combination chemotherapy the SVCS in small cell carcinoma can be effectively relieved. PMID- 7494239 TI - An unusual testicular metastasis from transitional cell carcinoma of the ureter: a case report. AB - Testicular tumor metastasis from transitional cell carcinoma (TCC) of the ureter is very rare. Herein, we report a case who presented with a palpable mass of left testis, after he underwent radical nephroureterectomy and bladder cuff excision and adjuvant radiotherapy and chemotherapy for the TCC of left ureter tumor for 5 months. We performed left radical orchiectomy and histology revealed a TCC identical to the original ureteral tumor. In a review of the literature, we found this case was the second case of metastatic testicular tumor from the ureteral TCC. In patients presenting the symptom of a palpable testicular mass, if they are known to have a ureteral TCC, metastatic neoplasm should be considered in the differential diagnosis, even if in the absence of other systemic metastasis. PMID- 7494240 TI - [A statistical analysis of oral squamous cell carcinoma]. AB - Oral squamous cell carcinoma is the most common malignant tumor of the oral cavity, and it accounts for about 90% of malignant oral tumors. We have retrospectively reviewed 211 patients with oral squamous cell carcinoma which were confirmed by histopathological examination at Kaohsiung Medical College Hospital in the period between 1987 and 1991. There were 194 males and 17 females, with a male to female ratio of 11.4 to 1. The age of the patients ranged from 23 to 95 years, mean 51.96 years, with the greatest incidence of disease in the ages between 40 and 59 years (63.3%). Squamous cell carcinoma occurred most often on the buccal mucosa. There were 40.8% of cases which occurred on the buccal mucosa, 25.6% on the tongue, 17.1% on the gingiva of mandible, 9.5% on the maxilla, 5.2% on the lower lip and 1.9% on the floor of mouth. 195 cases with a recorded history of habits in the patient's medical records were evaluated for the relationships between habits, patient's age and the location of the tumor involved. We have used a general factorial ANOVA to examine the relationship of habits and patients' age. A Hierarchical Loglinear Analysis was used to determine whether there was an association between habits and the location of tumor involved. Of the 195 cases, 144 (73.85%) chewed betel nut regularly. The mean age of the patients with betel nut chewing habits (49.04 years) was significantly younger than that of the patients without betel nut chewing habits (59.27 years) (p = 0.006).(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7494241 TI - [The changes of central and peripheral nerve conduction and the effect of thyroxine replacement in thyroidectomized rats]. AB - The aim of this study is to make serial BAEPs and PNCS in hypothyroid rats in order to provide objective and quantitative methods of detecting dysfunction of the central and peripheral nervous system in hypothyroid status and elucidating the relationship between the recovery potential and the duration of hypothyroid status. Thyroidectomy was performed in eighteen naive 5-month-old Sprague-Dawley rats in order to induce hypothyroid status, which was confirmed by RIA study 1-2 months after the surgery. Initial BAEP and PNCS were performed in two groups of the hypothyroid animals 1 and 3 months after thyroidectomy. Following the initial examination, thyroxin replacement therapy was given to each hypothyroid group, and then followed by BAEPs and PNCS at two month intervals, up to twice normal studies or six months after the initiation of therapy. In the BAEP study, the prolonged I-V inter-peak latency was the most consistent abnormal finding in all hypothyroid rats. Delayed peak latencies as well as prolonged I-III and III-V inter-peak intervals occurred when the hypothyroid status was longer than one month. We noted that the longer the hypothyroid status, the more severe the central conduction dysfunction. Nonetheless, these abnormalities always returned to normal after the replacement therapy if the hypothyroid state was shorter than 3 months in duration. For the PNCS study, all groups of thyroidectomized rats showed the normal results. We think the peripheral nervous system of rats may be more resistant to damage by hypothyroidism than the central nervous system. PMID- 7494242 TI - Human immunodeficiency virus type 1 infection of SK-N-MC cells: domains of gp120 involved in entry into a CD4-negative, galactosyl ceramide/3' sulfo-galactosyl ceramide-positive cell line. AB - The primary receptor for human immunodeficiency virus (HIV) is the CD4 molecule; however, in vitro evidence suggests that a neutral glycolipid, galactosyl ceramide (GalCer) or a derivative molecule, 3' sulfogalactosyl ceramide (GalS), may serve as an alternative receptor for HIV type 1 (HIV-1) in cells of neural and colonic origin. Biochemical studies have demonstrated that recombinant gp120 envelope protein binds to GalCer/GalS in both solid-phase enzyme-linked immunosorbent assay and high-performance thin-layer chromatography overlays. We have used the SK-N-MC cell line, a CD4-negative, GalCer/GalS-positive cell line previously characterized as susceptible to HIV-1 infection, to identify virus isolates with either a positive infection phenotype, HIVHxB2, or a negative infection phenotype, HIV-1(89.6). Using a solid-phase virus binding assay, we determined the level of restriction in HIV-1(89.6) infection to be at the level of virus-glycolipid binding. Furthermore, using HIV-1HxB2-HIV-1(89.6) chimeras, we have identified a 193-amino-acid fragment from the envelope region of HIV 1HxB2 containing the V3, V4, and V5 regions which confers a positive infection phenotype on the HIV-1(89.6) background. Recombinant viruses which separate this 193-amino-acid fragment into two distinct chimeras are each able to confer a positive infection phenotype on the background of HIV89.6, suggesting that a stable GalCer/GalS-envelope interaction is dependent on the conformation of the envelope protein in the context of the viral membrane. Alternatively, the GalCer/GalS-gp120 bond may involve multiple sites on the oligomeric envelope protein. PMID- 7494243 TI - Genesis of Sindbis virus by in vivo recombination of nonreplicative RNA precursors. AB - Genetically engineered RNA transcripts coding for various Sindbis virus (SIN) genes were used to study structure and sequence requirements of RNA recombination in BHK cells. Three different groups of RNA transcripts were made: (i) RNAs which retain the ability to replicate and which carry sequences coding for either viral polymerase or viral structural proteins; (ii) RNAs which lack the complete 3' end of the SIN genome and thus are incapable of replicating; and (iii) RNAs which lack the complete 5' end of the SIN genome and also are incapable of replicating. BHK cells were transfected with specific combinations of these precursor RNAs, and virus production and RNA synthetic abilities of the released virus were determined. We demonstrate in vivo generation of infectious SIN by fusion of (i) replicative RNAs to nonreplicative RNAs and (ii) two nonreplicative RNA precursors. Both homologous and nonhomologous types of recombinations were observed. In the homologous type of recombination, a 694-nucleotide overlap at the crossover region of the first pair of precursors resulted in the addition of an A residue converting the UAG stop codon of nonstructural protein P4 to a UAA stop codon. In the nonhomologous type of recombination, the crossover sites contained deletion of up to 76 nucleotides from one of the precursors and complete preservation of junction sequence from the other precursor. This is also the first report that a cytoplasmic RNA virus can be generated from biologically nonreplicative RNA precursors. These results have implications for initiation of viral RNA synthesis and recombination between RNA viral genomes in general. We favor template switching as a mechanism for the fusion events described here and suggest inclusion of polymerase scanning of diverse nonreplicative RNAs as an inherent feature of the copy choice model of RNA recombination. Very importantly, the facile nature of RNA recombination occurring between nonreplicative RNA precursors should speed up the production and analysis of targeted mutants of SIN and possibly other RNA viruses. PMID- 7494245 TI - Receptor-induced conformational changes in the subgroup A avian leukosis and sarcoma virus envelope glycoprotein. AB - We recently reported that Tva, the host cell receptor for subgroup A avian leukosis and sarcoma viruses, binds specifically to the subgroup A envelope glycoprotein (Env-A) (J.M. Gilbert, P. Bates, H. E. Varmus, and J. M. White, J. Virol. 68:5623-5628, 1994). Here we have tested the hypothesis that binding of Tva causes conformational changes in Env-A that correlate with its conversion from a fusion-inactive to a fusion-active state. Conformational changes were examined by both a proteolysis and an immunoprecipitation assay. A temperature dependent conformational change, demonstrated by the generation of a specific thermolysin digestion product of the surface (SU) subunit, occurred when a soluble form of Tva (sTva) was incubated with Env-A. sTva did not induce this conformational change in Env-C or in a noninfectious precursor form of Env-A, Env A CL. However sTva did induce the conformational change in Env-A CL that had been pretreated in vitro to produce the SU and transmembrane (TM) subunits. Moreover, interaction of Tva with Env-A at 25 degrees C, but not at 4 degrees C, appeared to reveal a previously buried segment of the putative fusion peptide of Env-A. Our results suggest that binding of Tva to Env-A results in specific conformational changes in the Env-A glycoprotein that are relevant to the activation of its fusion function. PMID- 7494244 TI - Adenovirus E1A243 disrupts the ATF/CREB-YY1 complex at the mouse c-fos promoter. AB - The adenovirus E1A243 protein can activate transcription of the mouse c-fos gene in a manner that depends on treatment of cells with inducers or analogs of cyclic AMP (cAMP). Activation requires conserved region 1 and the N-terminal domain of E1A243 and is mediated by a 22-bp E1A response element containing a cAMP response element (CRE) at -67 and a binding site for transcription factor YY1 at -54. In the absence of E1A243, YY1 represses CRE-dependent transcription of c-fos by physically interacting with ATF/CREB proteins bound to the -67 CRE. Here we present evidence that expression of E1A243 leads to relief of YY1-mediated repression by a disruption of the ATF/CREB-YY1 complex. Addition of E1A243 to in vitro binding assays prevented binding of ATF-2 to glutathione S-transferase-YY1. Similarly, expression of E1A243 in HeLa cells prevented the association of a YY1 VP16 fusion protein with endogenous ATF/CREB proteins bound to the -67 CRE of a transfected c-fosCAT reporter plasmid. In each case, the N-terminal domain of E1A243, which mediates a direct interaction with YY1, was responsible for disruption of the ATF/CREB-YY1 complex. On the basis of these and previously published results, we present a model for the synergistic transcriptional activation of the c-fos gene by E1A243 and cAMP. PMID- 7494247 TI - Discriminated selection among viral peptides with the appropriate anchor residues: implications for the size of the cytotoxic T-lymphocyte repertoire and control of viral infection. AB - Structural characterization of peptides restricted by major histocompatibility complex (MHC) class I molecules has identified residues critical for MHC class I binding and for T-cell receptor recognition. For example, optimal peptides fitting into the murine MHC class I Db groove are 9 to 11 amino acids long and require as MHC anchor residues an Asn (N) at position 5 and also either a hydrophobic residue, a Met (M) or a Cys (C), at the carboxy terminus. The three known Db-restricted peptides of lymphocytic choriomeningitis virus (LCMV) are glycoproteins GP1 (amino acids [aa] 33 KAVYNFATC), GP2 (aa 276 SGVENPGGYCL), and nucleoprotein NP (aa 396 FQPQNGQFI). In addition to these two GP and one NP peptides, computer search revealed 11 other GP peptide sequences and 20 additional NP sequences that contained the Db binding motif. By Db competitive binding analysis, only two of these 11 GP peptides and 1 of these 20 NP peptides bound to the MHC Db molecule with an affinity equivalent to the measured affinities for the three known GP1, GP2, and NP cytotoxic T-lymphocyte (CTL) epitopes. No CTL specific for these three peptides were generated when H-2b mice were inoculated with viral variants in which either the two known GP epitopes (GP1 and GP2; termed GPV) or the GPV and NP epitopes (termed GPV + NPV) were mutated. However, a novel CD8+ anti-LCMV CTL response ordinarily not seen in H-2b mice inoculated with wild-type virus was noted when such mice were inoculated with the GPV + NPV-mutated variant. This result indicates that (i) despite large numbers of peptides containing the appropriate anchor residues within a viral protein, only a restricted number induce CTL, thereby maintaining a limited CTL repertoire, (ii) despite the limited repertoire, the immune system retains the flexibility to generate an immune response(s) to a previously silent protein(s), suggesting a hierarchial control mechanism, and (iii) identification of a primary amino acid sequence is not sufficient, per se, to predict CTL epitopes, and peptide conformations are likely more complex than indicated by simple linear sequence comparisons. PMID- 7494246 TI - Immunodominance of major histocompatibility complex class I-restricted influenza virus epitopes can be influenced by the T-cell receptor repertoire. AB - We have used T-cell receptor beta-chain transgenic mice to determine the effects of a limited T-cell receptor repertoire on major histocompatibility complex class I-restricted epitope selection during the course of an influenza virus infection. Analysis of T-cell hybridomas generated from wild-type and transgenic mice demonstrated that the viral epitope recognized depended on the available T-cell receptor repertoire. Wild-type T-cell hybridomas recognized epitopes derived from the nucleoprotein and basic polymerase molecules, whereas hybridomas generated from transgenic mice recognized epitopes derived from the nonstructural protein and the matrix protein. There was no overlap in specificity between the two panels of hybridomas. This reciprocal pattern of specificity was also apparent in cytoxicity assays with brochoalveolar lavage cells isolated from the lungs of influenza virus-infected mice. T-cell receptor usage in the transgenic hybridomas was very restricted, with only one V alpha element used for ech of the two viral epitopes recognized. In the case of the hybridomas reactive to the nonstructural protein, sequence analysis showed that they all expressed V alpha 4J alpha 32 chains associated with the same junctional amino acids (Leu-Leu) that were encoded by five different nucleotide sequences, indicating a strong selection for T-cell receptor usage. Taken together, these data demonstrate that the available T-cell receptor repertoire can have a profound effect on the immunodominance of class I-restricted epitopes during a viral infection. PMID- 7494248 TI - High-titer packaging cells producing recombinant retroviruses resistant to human serum. AB - Novel retroviral protein expression constructs were designed to retain minimal retroviral sequences and to express dominant selectable markers by reinitiation of translation after expression of the viral genes. HT1080 cells were selected as producer cells for their ability to release high-titer viruses that are resistant to inactivation by human serum. Two HT1080-based packaging cell lines which produce Moloney murine leukemia virus cores with envelope glycoproteins of either amphotropic murine leukemia virus (FLYA13 line) or cat endogenous virus RD114 (FLYRD18 line) are described. Direct comparison with previous retroviral packaging systems indicated that 100-fold-higher titers of helper-free recombinant viruses were released by the FLYA13 and FLYRD18 lines. PMID- 7494249 TI - A Tat-induced auto-up-regulatory loop for superactivation of the human immunodeficiency virus type 1 promoter. AB - The virus-encoded Tat protein strongly activates transcription of human immunodeficiency virus (HIV). A well-recognized mechanism involves interaction of Tat with the nascent RNA transcript of the viral tar gene; mutation of tar greatly decreases activation by Tat. However, Tat still provides a low level of activation, demonstrating that it also has a tar-independent mode of action. We propose that this tar-independent mode of Tat action is through activation of gene transcription to produce tumor necrosis factor alpha. This cytokine and other compounds that activate NF-kappa B up-regulate the HIV promoter at a low level, similarly to the second Tat action. Through this mechanism, they also activate promoters of tumor necrosis factor alpha and other cytokines and thereby establish an auto-up-regulatory loop. Activated NF-kappa B motifs in the HIV promoter synergize with Tat/tar. Mutations of these motifs decrease activation by Tat to a few percent of the wild-type value. In cooperation, the two modes of activation by Tat (tar dependent and cytokine based) set up positive up regulatory loops which greatly superactivate transcription of HIV. Agents that block these synergistic pathways at three different steps and are more inhibitory in combination than is any one alone have been found. Thereby, multidrug modalities for transcription of HIV are proposed for virus suppression. PMID- 7494250 TI - Analysis of picornavirus 2A(pro) proteins: separation of proteinase from translation and replication functions. AB - The poliovirus (PV) genome was manipulated by replacing its 2A-encoding sequence with the corresponding sequence of coxsackie B4 virus (CBV4) or human rhinovirus type 2 (HRV2). In vitro translation of the resulting chimeric PV genomes revealed a normal cis-cleavage activity for both heterologous 2A(pro) proteinases in the chimeric PV polyproteins. However, only the genome containing the 2A-encoding sequence of CBV4 (PV/CBV4-2A) yielded viable virus in transfected cells, producing a mixture of large and small plaques on HeLa cell monolayers. The large plaque variants were found to contain single-amino-acid mutations at a specific site near the C terminus of the CBV4 2A(pro) protein. When the same single-amino acid mutations were directly introduced into the parental PV/CBV4-2A genome, chimeric viruses with a large-plaque phenotype and a wild-type PV-like growth pattern were obtained upon transfection, an observation demonstrating that these point mutations alone had a drastic effect on the growth of the PV/CBV4 chimeric virus. On the other hand, the chimeric genome containing the 2A-encoding sequence of HRV2 (PV/HRV2-2A) produced a null phenotype in transfected HeLa cells, although low-level replication of this chimeric genome was evident. We conclude that only 2A(pro) of the more closely related enterovirus CBV4 is able to functionally substitute for that of PV in vivo, and a subtle genetic modification of the CBV4 2A(pro) protein results in a drastic improvement in the growth of the chimeric PV/CBV4-2A virus. In addition, this chimeric cDNA approach enabled us to dissect multiple biological functions encoded by the 2A(pro) proteins. PMID- 7494251 TI - Monensin and nigericin prevent the inhibition of host translation by poliovirus, without affecting p220 cleavage. AB - Addition of monensin or nigericin after poliovirus entry into HeLa cells prevents the inhibition of host protein synthesis by poliovirus. The infected cells continue to synthesize cellular proteins at control levels for at least 8 h after infection in the presence of the ionophore. Cleavage of p220 (gamma subunit of eukaryotic initiation factor 4 [eIF-4 gamma]), a component of the translation initiation factor eIF-4F, occurs to the same extent in poliovirus-infected cells whether or not they are treated with monensin. Two hours after infection there is no detectable intact p220, but the cells continue to translate cellular mRNAs for several hours at levels similar to those in uninfected cells. Nigericin or monensin prevented the arrest of host translation at all the multiplicities of poliovirus infection tested. At high multiplicities of infection, an unprecedented situation was found: cells synthesized poliovirus and cellular proteins simultaneously. Superinfection of vesicular stomatitis virus-infected HeLa cells with poliovirus led to a profound inhibition of vesicular stomatitis virus protein synthesis, while nigericin partially prevented this blockade. Drastic inhibition of translation also took place in influenza virus-infected Vero cells treated with nigericin and infected with poliovirus. These findings suggest that the translation of newly synthesized mRNAs is dependent on the integrity of p220, while ongoing cellular protein synthesis does not require an intact p220. The target of ionophore action during the poliovirus life cycle was also investigated. Addition of nigericin at any time postinfection profoundly blocked the synthesis of virus RNA, whereas viral protein synthesis was not affected if nigericin was added at 4 h postinfection. These results agree well with previous findings indicating that inhibitors of phospholipid synthesis or vesicular traffic interfere with poliovirus genome replication. Therefore, the action of nigericin on the vesicular system may affect poliovirus RNA synthesis. In conclusion, monensin and nigericin are potent inhibitors of poliovirus genome replication that prevent the shutoff of host translation by poliovirus while still permitting cleavage of p220. PMID- 7494252 TI - The NS2 protein of hepatitis C virus is a transmembrane polypeptide. AB - The NS2 protein of hepatitis C virus (HCV) is released from its polyprotein precursor by two proteolytic cleavages. The N terminus of this protein is separated from the E2/p7 polypeptide by a cleavage thought to be mediated by signal peptidase, whereas the NS2-3 junction located at the C terminus is processed by a viral protease. To characterize the biogenesis of NS2 encoded by the BK strain of HCV, we have defined the minimal region of the polyprotein required for efficient cleavage at the NS2-3 site and analyzed the interaction of the mature polypeptide with the membrane of the endoplasmic reticulum (ER). We have observed that although cleavage can occur in vitro in the absence of microsomal membranes, synthesis of the polyprotein precursor in the presence of membranes greatly increases processing at this site. Furthermore, we show that the membrane dependency for efficient in vitro processing varies among different HCV strains and that host proteins located on the ER membrane, and in particular the signal recognition particle receptor, are required to sustain efficient proteolysis. By means of sedimentation analysis, protease protection assay, and site-directed mutagenesis, we also demonstrate that the NS2 protein derived from processing at the NS2-3 site is a transmembrane polypeptide, with the C terminus translocated in the lumen of the ER and the N terminus located in the cytosol. PMID- 7494253 TI - Redistribution of microtubules and Golgi apparatus in herpes simplex virus infected cells and their role in viral exocytosis. AB - Earlier studies have shown that the Golgi apparatus was fragmented and dispersed in herpes simplex virus 1-infected Vero and HEp-2 cells but not in human 143TK- cells, that the fragmentation and dispersal required viral functions expressed concurrently with or after the onset of DNA synthesis (G. Campadelli-Fiume, R. Brandimarti, C. Di Lazzaro, P. L. Ward, B. Roizman, and M. R. Torrisi, Proc. Natl. Acad. Sci. USA 90:2798-2802, 1993), and that in 143TK- cells, but not Vero or HEp-2 cells, infected with viral mutants lacking the UL20 gene virions were glycosylated and transported to extracellular space (J. D. Baines, P. L. Ward, G. Campadelli-Fiume, and B. Roizman, J. Virol. 65:6414-6424, 1991; E. Avitabile, P. L. Ward, C. Di Lazzaro, M. R. Torrisi, B. Roizman, and G. Campadelli-Fiume, J. Virol. 68:7397-7405, 1994). Experiments designed to elucidate the role of the microtubules and of intact or fragmented Golgi apparatus in the exocytosis of virions showed the following. (i) In all cell lines tested (Vero, 143TK-, BHK, and Hep-2) microtubules underwent fragmentation particularly evident at the cell periphery and then reorganized into bundles which circumvent the nucleus. This event was not affected by inhibitors of viral DNA synthesis. We conclude that redistribution of microtubules may be required but is not sufficient for the fragmentation and dispersal of the Golgi apparatus. (ii) In all infected cell lines tested, nocodazole caused fragmentation and dispersal of the Golgi and a far more extensive depolymerization of the microtubules than was seen in untreated, infected Vero or HEp-2 cells. Taxol precluded the depolymerization of the microtubules and fragmentation of the Golgi in both infected cell lines. Neither nocodazole nor taxol affected the exocytosis of infectious virus from Vero, HEp-2, or 143TK- cells infected with wild-type virus. We conclude that the effects of nocodazole or of taxol are dominant over the effects of viral infection in the cell lines tested and that viral exocytosis is independent of the organization of microtubules or of the integrity of the Golgi apparatus. Lastly, the data suggest that herpes simplex viruses have evolved an exocytic pathway for which the UL20 protein is a component required in some cells but not others and in which this protein does not merely compensate for the fragmentation and dispersal of the Golgi apparatus. PMID- 7494254 TI - Assembly and catalytic properties of retrovirus integrase-DNA complexes capable of efficiently performing concerted integration. AB - The in vitro assembly process for forming nucleoprotein complexes containing linear retrovirus-like DNA and integrase (IN) was investigated. Solution conditions that allowed avian myeloblastosis virus IN to efficiently pair two separate linear DNA fragments (each 487 bp in length) containing 3' OH recessed long terminal repeat termini were established. Pairing of the viral termini by IN during preincubation on ice permitted these nucleoprotein complexes to catalyze the concerted insertion of the two termini into a circular DNA target (full-site reaction), mimicking the in vivo reaction. The three major solution determinants were high concentrations of NaCl (0.33 M), 1,4-dioxane, and polyethylene glycol. The aprotic solvent dioxane (15%) was significantly better (sixfold) than 15% dimethyl sulfoxide for forming complexes capable of full-site rather than half site integration events. Half-site reactions by IN involved the insertion of a single donor terminus into circular pGEM. Although NaCl was essential for the efficient promotion of the concerted integration reaction, dioxane was necessary to prevent half-site reactions from occurring at high NaCl concentrations. Under optimal solution conditions, the concerted integration reaction was directly proportional to a sixfold range of IN. The complexes appeared not to turn over, and few half-site donor-donor molecules were produced. In the presence of 0.15 or 0.35 M NaCl, dioxane prevented efficient 3' OH trimming of a blunt-ended donor by IN, suggesting that the complexes formed by IN with blunt-ended donors were different from those formed with donors containing 3' OH recessed termini for strand transfer. The results suggest that IN alone was capable of protein-protein and protein-DNA interactions that efficiently promote the in vitro concerted integration reaction. PMID- 7494255 TI - The mismatched nucleotides in the 5'-terminal hairpin of minute virus of mice are required for efficient viral DNA replication. AB - The 5'-terminal sequence in the DNA of the parvovirus minute virus of mice (MVM) is a palindrome. It can form a hairpin, the stem of which is entirely base-paired except for three consecutive unpaired nucleotides which form a bubble. Since this structure is well conserved among different parvoviruses, we examined its importance for viral replication by generating MVM mutants with alterations in this region. A clone of MVMp DNA which contained the entire 3' end and more than half of the 5' palindrome was made. Although it lacked the sequence information to form a wild-type bubble, this DNA was infectious. On transfection into A9 fibroblasts, it gave rise to a virus (MVMs) which had a bubble in its 5' palindrome. The bubble consisted of four mismatched nucleotides in the same location as the unpaired nucleotides of the wild-type palindrome. Apparently, neighboring plasmid sequences were incorporated into the viral DNA, enabling formation of the mismatch. This observation suggested that a bubble is critical for growth of MVM but that its sequence is not. To find out whether MVM lacking a bubble in the 5' palindrome is viable, we made a second clone in which the plasmid sequences incorporated in MVMs were removed. Transfection of this DNA gave rise to a virus (MVMx) in which the nucleotides unpaired in the wild-type hairpin are now fully base-paired. Although MVMx can be propagated, it is defective in comparison with wild-type MVMp; it exhibited about a 50-fold-lower ratio of plaque-forming units to DNA content. In mixed infections, MVMp consistently outgrew the bubbleless MVMx. The rate of accumulation of DNA replication intermediates was lower for MVMx than for the wild-type virus. Quantitative analysis of the 5' termini of replicative form DNA suggested that the ability of MVMx to convert hairpin 5' termini to extended termini is impaired. In contrast, the virus with the altered bubble, MVMs, behaved like the wild-type MVMp in all the assays. We conclude that MVM lacking a bubble in its 5' terminal DNA hairpin is less infectious than and has a selective disadvantage compared with wild-type MVM. The nucleotide sequence of the bubble is not critical. We provide evidence that the presence of a bubble is necessary for efficient viral DNA replication. PMID- 7494256 TI - Human CD4+ T-cell repertoire of responses to influenza A virus hemagglutinin after recent natural infection. AB - The human CD4+ T-cell repertoire of responses to hemagglutinin (HA) of influenza virus A/Beijing/32/92 was examined 3 to 6 months after natural infection by using a panel of 16-mer peptides overlapping by 11 residues. Short-term CD4+ T-cell lines were derived by using full-length HAs of virus A/Beijing/32/92 from 12 unrelated, major histocompatibility complex (MHC) class I and II haplotyped adults with a history of influenza in November and December 1993 and from 6 adults with no history of influenza during the preceding 4 years but who responded to HA. In contrast to recent murine studies, the human CD4+ T-cell repertoire of responses was dominated by the recognition of highly conserved epitopes. The HA2 subunit, widely regarded as nonimmunogenic, induced strong responses in every donor. This resulted in functional cross-reactivity among influenza A viruses. Our study included one pair of unrelated donors expressing identical HLA DRB1 and DQB1 alleles and two pairs of donors sharing low resolution MHC class II types. These pairs responded to identical peptides; furthermore, clearly identifiable patterns of response were seen in donors sharing single class II haplotypes, irrespective of the presence of other alleles and exposure history. Two conserved regions which induced responses in 17 of 18 donors were identified (residues 295 to 328 and 407 to 442). Possible implications for cross-reactive T-cell vaccines are discussed. PMID- 7494257 TI - Naturally occurring accessory gene mutations lead to persistent human immunodeficiency virus type 1 infection of CD4-positive T cells. AB - Proviral DNA from cells surviving severe but transient cytopathic effects, mediated by infection with recombinant human immunodeficiency virus type 1 (HIV 1) carrying a single gene mutation at vif, vpr, or vpu, was characterized by use of HIV-1-specific primer pairs in a two-step PCR. Deletion mutations were detected in a region that spanned the vif and vpr open reading frames. Cloning and sequencing of the amplified DNA from this region revealed frequent large deletions in a limited number of nucleotide positions. Analyses of the deletions suggested that (i) genetic recombination, (ii) template-primer slippage, and (iii) misalignment of the growing point during reverse transcription of the HIV-1 genome might be the mechanisms that generated the mutations. Apart from the large deletions, smaller deletions that gave frameshift mutations in vif and/or vpr prevailed. In addition, cells infected with a triple mutant defective in vif, vpr, and vpu did not show any cytopathic effect. Thus, mutations generating multiple accessory gene defects during HIV-1 replication correlate with viral persistence and loss of cytopathogenicity. PMID- 7494258 TI - Complex formation between the NS3 serine-type proteinase of the hepatitis C virus and NS4A and its importance for polyprotein maturation. AB - Processing of the hepatitis C virus polyprotein is mediated by host cell signalases and at least two virally encoded proteinases. Of these, the serine type proteinase encompassing the amino-terminal one-third of NS3 is responsible for cleavage at the four sites carboxy terminal of NS3. The activity of this proteinase is modulated by NS4A, a 54-amino-acid polyprotein cleavage product essential for processing at the NS3/4A, NS4A/4B, and NS4B/5A sites and enhancing cleavage efficiency between NS5A and NS5B. Using the vaccinia virus-T7 hybrid system to express hepatitis C virus polypeptides in BHK-21 cells, we studied the role of NS4A in proteinase activation. We found that the NS3 proteinase and NS4A form a stable complex when expressed as a single polyprotein or as separate molecules. Results from deletion mapping show that the minimal NS4A domain required for proteinase activation is located in the center of NS4A between amino acids 1675 and 1686 of the polyprotein. Amino acid substitutions within this domain destabilizing the NS3-NS4A complex also impair trans cleavage at the NS4A dependent sites. Similarly, deletion of amino-terminal NS3 sequences impairs complex formation as well as cleavage at the NS4B/5A site but not at the NS4A independent NS5A/5B site. These results suggest that a stable NS3-NS4A interaction is important for cleavage at the NS4A-dependent sites and that amino terminal NS3 sequences and the central NS4A domain are directly involved in complex formation. PMID- 7494259 TI - Function of a 5'-end genomic RNA mutation that evolves during persistent mouse hepatitis virus infection in vitro. AB - Persistently infected cultures of DBT cells were established with mouse hepatitis virus strain A59 (MHV-A59), and the evolution of the MHV leader RNA and 5' end of the genome was studied through 119 days postinfection. Sequence analysis of independent clones demonstrated an overall mutation frequency approaching 1.2 x 10(-3) to 6.7 x 10(-3). The rate of fixation of mutations was about 1.2 x 10(-5) to 7.6 x 10(-5) per nucleotide (nt) per day. In contrast to finding in bovine coronavirus, the MHV leader RNA sequences were extremely stable and did not evolve significantly during persistent infection. Rather, a 5' untranslated region (UTR) A-to-G mutation at nt 77 in the genomic RNA emerged by day 56 and accumulated until 50 to 80% of the genome-length molecules retained the mutation by 119 days postinfection. Although other 5'-end mutations were noted, only the nt 77 mutation was significantly associated with viral persistence in vitro. Mutations were also found in the 5' end of the p28 coding region, but no specific alterations accumulated in genome-length molecules through 119 days postinfection. The 5' UTR nt 77 mutation resulted in an 18-amino-acid open reading frame (ORF) upstream of the ORF 1a AUG start site. By in vitro translation assays, the small ORF was not translated into detectable product but the mutation significantly enhanced translation of the downstream p28 ORF about 2.5-fold. Variant viruses, containing either the nt 77 A-to-G mutation (V16-ATG+) or wild-type sequences at this locus (V1-ATG-), were isolated at 119 days postinfection. The variant viruses replicated more efficiently than wild-type virus and were extremely cytolytic in DBT cells, suggesting that the A-to-G mutation did not encode a nonlytic or attenuated phenotype. Consistent with the in vitro translation results, a significant increase (approximately 3.5-fold) in p28 expression was also observed with the mutant virus (V16-ATG+) in DBT cells compared with that in wild-type controls. These data indicate that MHV persistence was significantly associated with mutation and evolution in the 5' end UTR which enhanced the translation of the ORF 1a and potentially ORF 1b polyproteins which function in virus transcription and replication. PMID- 7494260 TI - Novel retroviral vectors for efficient expression of the multidrug resistance (mdr-1) gene in early hematopoietic cells. AB - We present data that retroviral gene expression in early hematopoietic cells is subjected to transcriptional controls similar to those previously described for embryonic stem cells. Transient transfection experiments revealed that both the viral enhancer region in the U3 region of the long terminal repeat as well as a repressor element coincident with the primer binding site of Moloney leukemia viruses are limiting for expression in hematopoietic cells in a differentiation dependent manner. Within the group of Moloney leukemia virus-related viruses, only the myeloproliferative sarcoma virus showed high enhancer activity in myeloid (including erythroid) cells. In contrast, enhancer regions related to the Friend mink cell focus-forming viruses mediate much higher gene expression levels in both multipotent and lineage-committed myeloid cells. In addition, transcriptional repression related to sequences in the primer binding site of Moloney leukemia virus-derived vectors is also found in early hematopoietic cells and can be overcome by using the corresponding sequences of the murine embryonic stem cell virus. On the basis of these results, two types of novel retroviral hybrid vectors were developed; they combine the U3 regions of either the Friend mink cell focus-forming virus family or the myeloproliferative sarcoma virus with the primer binding site of the murine embryonic stem cell virus. When used to express the human multiple drug resistance gene, these vectors substantially improve protection to cytostatic drugs in transduced hematopoietic cell lines FDC Pmix, TF-1, and K-562 in comparison with Moloney leukemia virus-derived vectors presently used for the stem cell protection approach in somatic gene therapy. PMID- 7494261 TI - Convergent evolution within the V3 loop domain of human immunodeficiency virus type 1 in association with disease progression. AB - Phylogenetic analysis was used to study in vivo genetic variation of the V3 region of human immunodeficiency virus type 1 in relation to disease progression in six infants with vertically acquired human immunodeficiency virus type 1 infection. Nucleotide sequences from each infant formed a monophyletic group with similar average branch lengths separating the sets of sequences. In contrast to the star-shaped phylogeny characteristic of interinfant viral evolution, the shape of the phylogeny formed by sequences from the infants who developed AIDS tended to be linear. A computer program, DISTRATE, was written to analyze changes in DNA distance values over time. For the six infants, the rate of divergence from the initial variant was inversely correlated with CD4 cell counts averaged over the first 11 to 15 months of life (r = -0.87, P = 0.024). To uncover evolutionary relationships that might be dictated by protein structure and function, tree-building methods were applied to inferred amino acid sequences. Trees constructed from the full-length protein fragment (92 amino acids) showed that viruses from each infant formed a monophyletic group. Unexpectedly, V3 loop protein sequences (35 amino acids) that were found at later time points from the two infants who developed AIDS clustered together. Furthermore, these sequences uniquely shared amino acids that have been shown to confer a T-cell line tropic phenotype. The evolutionary pattern suggests that viruses from these infants with AIDS acquired similar and possibly more virulent phenotypes. PMID- 7494262 TI - High-resolution mapping of the human T-cell leukemia virus type 1 Rex-binding element by in vitro selection. AB - Interactions between the Rex protein of HTLV-1 and the genomic Rex-binding element (XBE) mediate the cytoplasmic transport of viral mRNAs. However, it is uncertain which RNA sequences and structures contribute to Rex recognition. A portion of the viral genome that spanned the XBE was partially randomized, and functional Rex-binding variants were selected. Alignment of selected Rex-binding sequences revealed positions that were functionally conserved between different molecules. A model is presented in which a subset of the selected residues are in direct contact with Rex. Positions that covaried with one another were also found. These covariations support a secondary-structural model in which a central paired stem is symmetrically flanked by two bulge loops. On the basis of this model, site-directed mutations of the XBE were constructed and each half molecule was found to bind independently to Rex. The functional residues and secondary structures in the XBE half molecules bear a remarkable resemblance to the transactivation response region element of HIV-1. Since the transactivation response region element is known to interact specifically with arginine residues in the Tat protein, these results suggest that the XBE binds to the arginine-rich RNA-binding domain of Rex in a similar manner. This model is supported by the selection data. PMID- 7494265 TI - Accumulation of proteinase K-resistant prion protein (PrP) is restricted by the expression level of normal PrP in mice inoculated with a mouse-adapted strain of the Creutzfeldt-Jakob disease agent. AB - Creutzfeldt-Jakob disease (CJD) is a transmissible neurodegenerative disease of humans caused by an unidentified infectious agent, the prion. To determine whether there was an involvement of the host-encoded prion protein (PrPc) in CJD development and prion propagation, mice heterozygous (PrP+/-) or homozygous (PrP /-) for a disrupted PrP gene were established and inoculated with the mouse adapted CJD agent. In keeping with findings of previous studies using other lines of PrP-less mice inoculated with scrapie agents, no PrP-/- mice showed any sign of the disease for 460 days after inoculation, while all of the PrP+/- and control PrP+/+ mice developed CJD-like symptoms and died. The incubation period for PrP+/- mice, 259 +/- 27 days, was much longer than that for PrP+/+ mice, 138 +/- 12 days. Propagation of the prion was barely detectable in the brains of PrP /- mice and was estimated to be at a level at least 4 orders of magnitude lower than that in PrP+/+ mice. These findings indicate that PrPc is necessary for both the development of the disease and propagation of the prion in the inoculated mice. The proteinase-resistant PrP (PrPres) was undetectable in the brain tissues of the inoculated PrP-/- mice, while it accumulated in the affected brains of PrP+/+ and PrP+/- mice. Interestingly, the maximum level of PrPres in the brains of PrP+/- mice was about half of the level in the similarly affected brains of PrP+/+ mice, indicating that PrPres accumulation is restricted by the level of PrPc. PMID- 7494263 TI - Sequences flanking the pentanucleotide T-antigen binding sites in the polyomavirus core origin help determine selectivity of DNA replication. AB - Replication of the genomes of the polyomaviruses requires two virus-specified elements, the cis-acting origin of DNA replication, with its auxiliary DNA elements, and the trans-acting viral large tumor antigen (T antigen). Appropriate interactions between them initiate the assembly of a replication complex which, together with cellular proteins, is responsible for primer synthesis and DNA chain elongation. The organization of cis-acting elements within the origins of the polyomaviruses which replicate in mammalian cells is conserved; however, these origins are sufficiently distinct that the T antigen of one virus may function inefficiently or not at all to initiate replication at the origin of another virus. We have studied the basis for such replication selectivity between the murine polyomavirus T antigen and the primate lymphotropic polyomavirus origin. The murine polyomavirus T antigen is capable of carrying out the early steps of the assembly of an initiation complex at the lymphotropic papovavirus origin, including binding to and deformation of origin sequences in vitro. However, the T antigen inefficiently unwinds the origin, and unwinding is influenced by sequences flanking the T antigen pentanucleotide binding sites on the late side of the viral core origin. These same sequences contribute to the replication selectivity observed in vivo and in vitro, suggesting that the inefficient unwinding is the cause of the replication defect. These observations suggest a mechanism by which origins of DNA replication can evolve replication selectivity and by which the function of diverse cellular origins might be temporally activated during the S phase of the eukaryotic cell cycle. PMID- 7494264 TI - Murine polyomavirus and simian virus 40 large T antigens produce different structural alterations in viral origin DNA. AB - Murine polyomavirus (Py) and simian virus (SV40) encode homologous large T antigens (T Ags) and also have comparable sequence motifs in their core replication origins. While the ability of SV40 T Ag to produce specific distortions within the SV40 core replication origin (ori) in a nucleotide dependent fashion has been well documented, little is known about related effects of Py T Ag on Py ori DNA. Therefore, we have examined viral origin DNA binding in the presence of nucleotide and the resulting structural changes induced by Py and SV40 T Ags by DNase I footprinting and KMnO4 modification assays. The structural changes in the Py ori induced by Py T Ag included sites within both the A/T and early side of the core origin region, consistent with what has been shown for SV40. Interestingly, however, Py T Ag also produced sites of distortion within the center of the origin palindrome and at several sites within both the early and late regions that flank the core ori. Thus, Py T Ag produces a more extensive and substantially different pattern of KMnO4 modification sites than does SV40 T Ag. We also observed that both T Ags incompletely protected and distorted the reciprocal ori region. Therefore, significant differences in the interactions of Py and SV40 T Ags with ori DNA may account for the failure of each T Ag to support replication of the reciprocal ori DNA in permissive cell extracts. PMID- 7494266 TI - Hepatitis D virus RNA editing: specific modification of adenosine in the antigenomic RNA. AB - RNA editing plays a central role in the life cycle of hepatitis D virus (HDV), a subviral human pathogen. Previous studies (J.L. Casey, K.F. Bergmann, T.L. Brown, and J.L. Gerin, Proc. Natl. Acad. Sci USA 89:7149-7153, 1992; H. Zheng, T.-B. Fu, D. Lazinski, and J. Taylor, J. Virol. 66:4693-4697, 1992) had concluded that the genomic RNA of HDV was the target for RNA editing and that the editing reaction was a conversion of U to C. However, we show here that the antigenomic RNA of HDV is in fact the target for HDV RNA editing, which is therefore a conversion of A to G. This result is verified by using an assay specific for editing on the antigenomic RNA and by analyzing the editing of site-directed mutant RNAs in transfected cells and in cell extracts. Because editing occurs in the absence of viral antigens and the specificity for the HDV editing target site is present even in extracts from Drosophila cells, it is likely that HDV RNA is edited by one or more cellular factors that are conserved among higher eukaryotes. These results raise the likelihood that double-stranded RNA adenosine deaminase specifically edits HDV antigenomic RNA. PMID- 7494268 TI - Both the changes of six amino acids and the C-terminal truncation caused by a one base insertion in the defective env gene of Friend spleen focus-forming virus significantly affect the pathogenic activity of the encoded leukemogenic membrane glycoprotein (gp55). AB - Friend spleen focus-forming virus (F-SFFV) causes acute erythroleukemia in mice and encodes in its defective env gene an Env-like membrane glycoprotein (gp55). The F-SFFV env gene has three characteristic structures compared with that of ecotropic murine leukemia viruses (MuLVs): substitution by the polytropic MuLV env sequence, a 585-bp deletion, and a 1-bp insertion. All of these characteristic structures are essential for the leukemogenic potential of gp55 of polycythemia-inducing isolates of F-SFFV (F-SFFVp). The 1-bp insertion causes changes of six amino acids and truncation by 34 amino acids at the C terminus. In this study, we constructed 12 mutant F-SFFV genomes starting from the wild-type F SFFVp and examined the effect of the C-terminal truncation and the six altered amino acids on the pathogenic activity of gp55. The results indicated that at least 18 to 24 amino acids must be deleted from the C terminus for the env product to be pathogenically active. We also found that the six altered amino acids contributed significantly to the pathogenic activity of gp55. Analyses of the cellular processing of these mutant gp55s supported a correlation between the pathogenic activity of gp55 and its efficiency in overall cellular processing. PMID- 7494267 TI - Role of mutations G-480 and C-6203 in the attenuation phenotype of Sabin type 1 poliovirus. AB - Of the 55 point mutations which distinguish the type 1 poliovirus vaccine strain (Sabin 1) from its neurovirulent progenitor (P1/Mahoney), two have been strongly implicated by previous studies as determinants of the attenuation phenotype. A change of an A to a G at position 480, located within the 5' noncoding region, has been suggested to be the major attenuating mutation, analogous to the mutations at positions 481 and 472 in poliovirus types 2 and 3, respectively. In addition, the change of a U to a C at position 6203, resulting in an amino acid change in the polymerase protein 3D, has also been implicated as a determinant of attenuation, albeit to a lesser extent. To assess the contributions of these mutations to attenuation and temperature sensitivity, reciprocal changes were generated at these positions in infectious cDNA clones of Sabin 1 and P1/Mahoney. Assays in tissue culture and primates indicated that the two mutations make some contribution to the temperature sensitivity of the Sabin 1 strain but that neither is a strong determinant of attenuation. PMID- 7494269 TI - Binding of cellular repressor protein or the IE2 protein to a cis-acting negative regulatory element upstream of a human cytomegalovirus early promoter. AB - We have previously shown that the human cytomegalovirus early UL4 promoter has upstream negative and positive cis-acting regulatory elements. In the absence of the upstream negative regulatory region, the positive element confers strong transcriptional activity. The positive element contains a CCAAT box dyad symmetry and binds the cellular transcription factor NF-Y. The effect of the negative regulatory element is negated by the viral IE2 protein (L. Huang, C.L. Malone, and M.F. Stinski, J. Virol. 68:2108, 1994). We investigated the binding of cellular or viral IE2 protein to the negative regulatory region. The major cis acting negative regulatory element was located between -168 and -134 bp relative to the transcription start site. This element could be transferred to a heterologous promoter, and it functioned in either orientation. Mutational analysis demonstrated that a core DNA sequence in the cis-acting negative regulatory element, 5'-GTTTGGAATCGTT-3', was required for the binding of either a cellular repressor protein(s) or the viral IE2 protein. The cellular DNA binding activity was present in both nonpermissive HeLa and permissive human fibroblast cells but more abundant in HeLa cells. Binding of the cellular repressor protein to the upstream cis-acting negative regulatory element correlates with repression of transcription from the early UL4 promoter. Binding of the viral IE2 protein correlates with negation of the repressive effect. PMID- 7494270 TI - Altered cytokine expression in T lymphocytes from human immunodeficiency virus Tat transgenic mice. AB - Examination of the interaction between human immunodeficiency virus (HIV) regulatory gene products and the host immune system is fundamental to understanding the pathogenesis of HIV and could reveal possible targets for therapeutic intervention in the treatment of AIDS. The HIV Tat gene is a potential candidate for this type of strategy. Transgenic mice can be used to investigate the in vivo effects of Tat on the developing and dynamic immune system and on cellular gene expression. Thus, we have generated transgenic mice that harbor the HIV type 1 Tat gene under the transcriptional control of the human CD2 gene regulatory elements. This expression cassette results in high level, tissue-specific transcription of the transgene within the T-cell compartment. In this report, we demonstrate the effects of Tat on the in vivo immune system. CD2-Tat transgenic mice show no signs of aberrant thymic development and have normal levels of T-cell subsets in the thymus and peripheral lymphoid organs. However, activated T cells from transgenic mice contain increased levels of tumor necrosis factor beta mRNA as well as biologically active tumor necrosis factor protein and express elevated levels of transforming growth factor beta and interleukin-4 receptor mRNA. These increased cytokine levels do not appear to alter mitogen- or antigen-stimulated responses or induce the formation of dermal lesions in ageing mice. Such investigations should provide insight into the combination of host immune factors mediating pathogenesis in HIV infection. PMID- 7494271 TI - The Vif protein of human and simian immunodeficiency viruses is packaged into virions and associates with viral core structures. AB - The vif gene of human and simian immunodeficiency viruses (HIV and SIV) encodes a late gene product that is essential for viral infectivity in natural target cells. Virions produced in the absence of Vif are abnormal in their ultrastructural morphology and are severely impaired in the ability to complete proviral DNA synthesis upon entry into new target cells. Because previous studies failed to detect Vif protein in virus particles, Vif is believed to influence virus infectivity indirectly, by affecting virion assembly, release, and/or maturation. In this report, we reexamined the possibility that Vif is a virion associated protein. Utilizing high-titer Vif-specific antibodies, a sensitive immunoblot technique, and highly concentrated virus preparations, we detected a 23-kDa Vif-reactive protein in wild-type HIV type 1 (HIV-1) and a 27-kDa Vif reactive protein in wild-type SIVSM virions. Neither protein was present in virions derived from vif-deficient HIV-1 and SIVSM proviral constructs. Vif protein content was similar among different strains of HIV-1 and was independent of the cell type (permissive or nonpermissive) used to produce the virus. To determine the subvirion localization of Vif, HIV-1 virions were treated with proteinase K or Triton X-100 to remove virion surface proteins and the viral membrane, respectively, purified through sucrose, and analyzed by immunoblot analysis. Vif protein content was not affected by the removal of external surface proteins or by the removal of the viral membrane and submembrane p17Gag matrix protein. Instead, Vif colocalized with viral core structures which sedimented at a density of 1.25 g/ml on linear sucrose gradients (enveloped HIV-1 particles sediment at a density of 1.17 g/ml). Finally, the amount of Vif protein packaged into virions was estimated to be on the order of 1 molecule of Vif for every 20 to 30 molecules of p24Gag, or between 60 and 100 molecules of Vif per particle. These results indicate that Vif represents an integral component of HIV and SIV particles and raise the possibility that it plays a direct role in early replication events. PMID- 7494272 TI - Oncogenicity of human papillomavirus- or adenovirus-transformed cells correlates with resistance to lysis by natural killer cells. AB - The reasons for the dissimilar oncogenicities of human adenoviruses and human papillomaviruses (HPV) in humans are unknown but may relate to differences in the capacities of the E1A and E7 proteins to target cells for rejection by the host natural killer (NK) cell response. As one test of this hypothesis, we compared the abilities of E1A- and E7-expressing human fibroblastic or keratinocyte derived human cells to be selectively killed by either unstimulated or interferon (IFN)-activated NK cells. Cells expressing the E1A oncoprotein were selectively killed by unstimulated NK cells, while the same parental cells but expressing the HPV type 16 (HPV-16) or HPV-18 E7 oncoprotein were resistant to NK cell lysis. The ability of IFN-activated NK cells to selectively kill virally transformed cells depends on IFN's ability to induce resistance to NK cell lysis in normal (i.e., non-viral oncogene-expressing) but not virally transformed cells. E1A blocked IFN's induction of cytolytic resistance, resulting in the selective lysis of adenovirus-transformed cells by IFN-activated NK cells. The extent of IFN induced NK cell killing of E1A-expressing cells was proportional to the level of E1A expression and correlated with the ability of E1A to block IFN-stimulated gene expression in target cells. In contrast, E7 blocked neither IFN-stimulated gene expression nor IFN's induction of cytolytic resistance, thereby precluding the selective lysis of HPV-transformed cells by IFN-activated NK cells. In conclusion, E1A expression marks cells for destruction by the host NK cell response, whereas the E7 oncoprotein lacks this activity. PMID- 7494274 TI - The phenotype in vitro and in infected cells of herpes simplex virus 1 alpha trans-inducing factor (VP16) carrying temperature-sensitive mutations introduced by substitution of cysteines. AB - alpha trans-inducing factor (alpha TIF, VP16, Vmw65) is an essential structural protein of herpes simplex virus, being required for virion assembly. The protein also forms complexes with host proteins and a response element and transactivates the alpha genes which carry this element. The protein contains an acidic carboxyl terminus required for transactivation and a much larger amino-terminal domain required for promoter recognition. We report the first set of temperature sensitive (ts) mutations deliberately introduced into the protein by substitution of the cysteine codons with those specifying glycine at positions 78, 102, and 176, either singly or in combinations. We report the following results. (i) All mutated proteins synthesized in vitro formed complexes with the DNA response element at room temperature. However, the mutant with the triple substitution and two mutants with substitutions in two of the three cysteines exhibited a ts phenotype at 33 and 37 degrees C, and one exhibited a ts phenotype only at 37 degrees C. (ii) Replacement of wild-type alpha TIF with genes carrying substitutions in any two cysteines conferred a ts phenotype for replication at 39.5 degrees C. Shift-down experiments indicated that the 10(4)- to 10(5)-fold reduction in virus yield at the nonpermissive temperature was due to the disfunction of alpha TIF late in infection, presumably in virion maturation. (iii) The alpha TIF expressed in cells infected with mutant viruses exhibited the same ts phenotype in protein-DNA complex formation as those expressed in vitro from mutated plasmids. Although the virus carrying the alpha TIF substitutions at Cys-102 and Cys-176 failed to induce a reporter gene linked to the alpha 4 promoter at 39.5 degrees C, it replicated as well as the parent virus in cells maintained for the first 10 h of infection at 39.5 degrees C. We conclude the following. (i) Formation of DNA-protein complexes containing alpha TIF is a poor prognosticator of alpha TIF function. (ii) The data presented here and in the literature strongly support the hypothesis that the secondary structure of the alpha TIF is very sensitive to deletions or insertions which probably affect the interaction of alpha TIF with both viral proteins in the virion and cellular proteins during infection. As a consequence, deletion-insertion mutagenesis may not shed useful information on the role of transactivating function of alpha TIF in infection. (iii) Since cysteines may play a role in stabilizing the secondary structure of proteins, substitutions of cysteines may be a powerful technique for site-specific construction of ts mutants in essential viral proteins. PMID- 7494273 TI - Identification and characterization of ZIIBC, a complex formed by cellular factors and the ZII site of the Epstein-Barr virus BZLF1 promoter. AB - The transition from latency to lytic Epstein-Barr virus replication is dependent on the Epstein-Barr virus BZLF1 gene product. Genetic and biochemical attempts to link cellular second-messenger signaling pathways that trigger this transition with the subsequent viral gene cascade have identified functional elements within the BZLF1 promoter (Zp) that appear to bind undefined cellular transcription factors. One of these previously identified sites, ZII, has homology to consensus AP-1 and CREB binding sites, implying a role for these factors in the inductive process. We have identified and characterized ZIIBC, a ZII site binding complex that is distinct from the factors previously proposed to bind this site. Active ZIIBC was found to be present in both uninduced and chemically induced cell extracts at approximately equivalent concentrations. Analysis of the DNA sequence requirements for the binding of ZIIBC to the ZII site shows that sequences homologous to AP-1 and CREB consensus sites are necessary but not sufficient for complex formation. Although the components of ZIIBC that directly contact DNA were found to be of the same molecular masses (26 and 36 kDa) in both uninduced and chemically induced cell extracts, a slight mobility difference between DNA protein complexes formed by these two types of extracts is observable and indicates that ZIIBC is directly affected by chemical induction. The effects of ZIIBC binding to the ZII site on expression from Zp were evaluated, and they suggest that ZIIBC plays a critical role in the regulation of Zp expression. PMID- 7494275 TI - A mimotope from a solid-phase peptide library induces a measles virus neutralizing and protective antibody response. AB - A solid-phase 8-mer random combinatorial peptide library was used to generate a panel of mimotopes of an epitope recognized by a monoclonal antibody to the F protein of measles virus (MV). An inhibition immunoassay was used to show that these peptides were bound by the monoclonal antibody with different affinities. BALB/c mice were coimmunized with the individual mimotopes and a T-helper epitope peptide (from MV fusion protein), and the reactivity of the induced anti-mimotope antibodies with the corresponding peptides and with MV was determined. The affinities of the antibodies with the homologous peptides ranged from 8.9 x 10(5) to 4.4 x 10(7) liters/mol. However, only one of the anti-mimotope antibodies cross-reacted with MV in an enzyme-linked immunosorbent assay and inhibited MV plaque formation. Coimmunization of mice with this mimotope and the T-helper epitope peptide induced an antibody response which conferred protection against fatal encephalitis induced following challenge with MV and with the structurally related canine distemper virus. These results indicate that peptide libraries can be used to identify mimotopes of conformational epitopes and that appropriate immunization with these mimotopes can induce protective antibody responses. PMID- 7494276 TI - Mouse adenovirus type 1 causes a fatal hemorrhagic encephalomyelitis in adult C57BL/6 but not BALB/c mice. AB - Mouse adenovirus type 1 (MAV-1) produces a lethal disease in newborn or suckling mice characterized by infectious virus and viral lesions in multiple organs. Previous reports of MAV-1 infection of adult mice generally described serologic evidence of infection without morbidity or mortality. However, our current results demonstrate that MAV-1 causes a fatal illness in adult C57BL/6(B6) mice (50% lethal dose, [LD50], 10(3.0) PFU) but not in adult BALB/c mice at all of the doses tested (LD50, > or = 10(5.0) PFU). Adult (BALB/c x B6)F1 mice were intermediately susceptible (LD50, 10(4.5) PFU). Clinically, the sensitive B6 mice showed symptoms of acute central nervous system (CNS) disease, including tremors, seizures, ataxia, and paralysis. Light microscopic examination of CNS tissue from the B6 animals revealed petechial hemorrhages, edema, neovascularization, and mild inflammation in the brain and spinal cord. Analysis by electron microscopy showed evidence of inflammation, such as activated microglia, as well as swollen astrocytic endfeet and perivascular lipid deposition indicative of blood-brain barrier dysfunction. Outside of the CNS, the only significant pathological findings were foci of cytolysis in the splenic white pulp. Assessment of viral replication from multiple tissues was performed by using RNase protection assays with an antisense MAV-1 early region 1a probe. The greatest amounts of viral mRNA in MAV-1-infected B6 animals were located in the brain and spinal cord. Less viral message was detected in the spleen, lungs, and heart. No viral mRNA was detected in BALB/c mouse tissue, with the exception of low levels in the heart. Viral titers of organ tissues were also determined and were concordant with RNase protection findings on the brain and spinal cord but failed to demonstrate significant infectious virus in additional organs. Our experiments demonstrate that MAV-1 has a striking tropism for the CNS that is strain dependent, and this provides an informative in vivo model for the study of adenoviral pathogenesis. PMID- 7494277 TI - Replication efficiency of bovine papillomavirus type 1 DNA depends on cis-acting sequences distinct from the replication origin. AB - The viral elements required for the initiation of replication of bovine papillomavirus type 1 DNA include the origin region and two trans-acting factors, the E1 and E2 proteins. We now report that the replication efficiency of a DNA molecule which contains these three elements is modulated by other viral sequences. By measuring the extent of replication of deleted viral genomes in transfected mouse cells, we identified sequences required for maximal efficiency. Addition of these sequences to a construct carrying only the minimal origin region increased its replication. Among these cis-active elements, we identified a 69-bp fragment (nucleotides 4921 to 4990) which contains at least two binding sites for cellular proteins. One of them is the murine protein termed CDEBP, which recognizes the octameric motif ATCACGTG, identical to the yeast CDEI element. Either deletions affecting this CDEI box or a point mutation which impairs binding of CDEBP markedly decreased the extent of viral DNA replication. They had no detectable effect on viral transcription. PMID- 7494279 TI - Augmentation of virus secretion by the human immunodeficiency virus type 1 Vpu protein is cell type independent and occurs in cultured human primary macrophages and lymphocytes. AB - The human immunodeficiency virus type 1-specific Vpu protein is a small integral membrane phosphoprotein that induces degradation of the virus receptor CD4 in the endoplasmic reticulum and, independently, increases the release of progeny virions from infected cells. To address the importance of Vpu for virus replication in primary human cells such as peripheral blood mononuclear cells (PBMC) and monocyte-derived macrophages (MDM), we used three different sets of monocyte-tropic molecular clones of human immunodeficiency virus type 1: a primary isolate, AD8+, and two chimeric variants of the T-cell-tropic isolate NL4 3 carrying the env determinants of either AD8+ or SF162 monocyte-tropic primary isolates. Isogenic variants of these chimeric viruses were constructed to express either wild-type Vpu or various mutants of Vpu. The effects of these mutations in the vpu gene on virus particle secretion from infected MDM or PBMC were assessed by determination of the release of virion-associated reverse transcriptase into culture supernatants, Western blot (immunoblot) analysis of pelleted virions, and steady-state or pulse-chase metabolic labeling. Wild-type Vpu increased virus release four- to sixfold in MDM and two- to threefold in PBMC, while nonphosphorylated Vpu and a C-terminal truncation mutant of Vpu were partially active on virus release in primary cells. These results demonstrate that Vpu regulates virus release in primary lymphocyte and macrophage cultures in a similar manner and to a similar extent to those previously observed in HeLa cells or CD4+ T-cell lines. Thus, our findings provide evidence that Vpu functions in a variety of human cells, both primary cells and continuous cell lines, and mutations in Vpu affect its biological activity independent of the cell type and virus isolate used. PMID- 7494278 TI - A rabbitpox virus serpin gene controls host range by inhibiting apoptosis in restrictive cells. AB - Poxviruses are unique among viruses in encoding members of the serine proteinase inhibitor (serpin) superfamily. Orthopoxviruses contain three serpins, designated SPI-1, SPI-2, and SPI-3. SPI-1 encodes a 40-kDa protein that is required for the replication of rabbitpox virus (RPV) in PK-15 or A549 cells in culture (A. N. Ali, P. C. Turner, M. A. Brooks, and R. W. Moyer, Virology 202:305-314, 1994). Examination of nonpermissive human A549 cells infected with an RPV mutant disrupted in the SPI-1 gene (RPV delta SPI-1) suggests there are no gross defects in protein or DNA synthesis. The proteolytic processing of late viral structural proteins, a feature of orthopoxvirus infections associated with the maturation of virus particles, also appears relatively normal. However, very few mature virus particles of any kind are produced compared with the level found in infections with wild-type RPV. Morphological examination of RPV delta SPI-1-infected A549 cells, together with an observed fragmentation of cellular DNA, suggests that the host range defect is associated with the onset of apoptosis. Apoptosis is seen only in RPV delta SPI-1 infection of nonpermissive (A549 or PK-15) cells and is absent in all wild-type RPV infections and RPV delta SPI-2 mutant infections examined to date. Although the SPI-1 gene is expressed early, before DNA replication, the triggering apoptotic event occurs late in the infection, as RPV delta SPI-1-infected A549 cells do not undergo apoptosis when infections are carried out in the presence of cytosine arabinoside. While the SPI-2 (crmA) gene, when transfected into cells, has been shown to inhibit apoptosis, our experiments provide the first indication that a poxvirus serpin protein can inhibit apoptosis during a poxvirus infection. PMID- 7494280 TI - Antibody-forming cell response to virus challenge in mice immunized with DNA encoding the influenza virus hemagglutinin. AB - Immunization of mice with DNA encoding the influenza virus hemagglutinin (HA) affords complete protection against lethal influenza virus infection and the means to investigate the mechanisms of B-cell responsiveness to virus challenge. Using a single-cell enzyme-linked immunospot assay, we sought to determine the localization of HA-specific antibody-forming cells (AFCs) during the development of humoral immunity in mice given HA DNA vaccine by gene gun. At 33 days postvaccination, populations of AFCs were maintained in the spleen and bone marrow. In response to lethal challenge with influenza virus, the AFCs became localized at the site of antigenic challenge, i.e., within the draining lymph nodes of the lung compartment. Immunoglobulin G (IgG)- and IgA-producing AFCs were detected in lymph nodes of the upper and lower respiratory tracts, underscoring their importance in clearing virus from the lungs. Response to challenge required competent CD4+ T cells, without which no AFCs were generated, even those producing IgM. By contrast, in mice vaccinated with an HA-containing subunit vaccine, fewer AFCs were generated in response to challenge, and these animals were less capable of resisting infection. Our findings demonstrate the comparable localization of AFCs in response to challenge in mice vaccinated with either HA DNA or live virus. Moreover, the former strategy generates both IgG- and IgA-producing plasma cells. PMID- 7494281 TI - Cooperative binding of multimeric phosphoprotein (P) of vesicular stomatitis virus to polymerase (L) and template: pathways of assembly. AB - It was previously shown that the phosphoprotein (P) of vesicular stomatitis virus must undergo phosphorylation-dependent multimerization to become transcriptionally active. Phosphorylation at S-60 and/or T-62 by casein kinase II or substitution of these residues by D is required for multimer formation. We now find that substitution of either one of these residues by A prevents phosphorylation by casein kinase II and multimer formation. The binding of multimeric P to the other two transcriptional components of vesicular stomatitis virus (L protein and the N-RNA template) has been characterized by using P immobilized on beads through its poly(His) tag to facilitate recovery of bound complexes. Multimerization of P was absolutely required for binding to both L and template. Multimeric P combined with the polymerase enzyme (L) in a stoichiometric 1:1 complex, which bound to the N-RNA template much more strongly than multimeric P alone. Substitution of S-227 and S-233 by A residues had no effect on multimerization or binding of L to P but prevented binding of both P and L to template and abolished transcriptional activity. In contrast, substitution of these residues with D residues had no effect on template binding or activity. However, substitution at these sites by either D or A largely abolished phosphorylation by L-associated kinases, thus identifying S-227 and S 233 as the major sites targeted by these kinases and confirming that phosphorylation of P protein by L-associated kinases is without transcriptional effect. PMID- 7494283 TI - Expression of the polyomavirus minor capsid proteins VP2 and VP3 in Escherichia coli: in vitro interactions with recombinant VP1 capsomeres. AB - The polyomavirus VP2 and VP3 capsid proteins were expressed in Escherichia coli. The majority of the expressed proteins were in an insoluble fraction, and they were extracted and initially purified in 8 M urea before renaturation. Soluble VP2 and VP3 were mixed with purified recombinant VP1 capsomeres, and their interactions were assayed by immunoprecipitation and ion-exchange chromatography. Coimmunoprecipitation could be demonstrated with antibodies to either VP1 or VP2/VP3. Mixing recombinant VP1 with VP2 and VP3 modified the recognition of VP1 by domain-specific antipeptide antibodies and altered the chromatographic behavior of the individual proteins. Similar results were observed when a truncated VP1 protein, delta NCOVP1, with 62 amino acids deleted from the carboxy terminus was mixed with VP2/VP3. After the mixing, equilibrium dissociation constants for their binding to either VP1 or delta NCOVP1 were determined to be 0.37 +/- 0.23 microM for VP2 and 0.18 +/- 0.21 microM for VP3. These studies demonstrate that the recombinant VP2 and VP3 proteins interact with VP1 to affect the biochemical properties of VP1 capsomeres and to change the epitope accessibility of VP1 pentamers. These changes may reflect conformational alterations in VP1 capsomeres which are necessary for viral genome encapsidation. PMID- 7494282 TI - Branchpoint and polypyrimidine tract mutations mediating the loss and partial recovery of the Moloney murine sarcoma virus MuSVts110 thermosensitive splicing phenotype. AB - Balanced splicing of retroviral RNAs is mediated by weak signals at the 3' splice site (ss) acting in concert with other cis elements. Moloney murine sarcoma virus MuSVts110 shows a similar balance between unspliced and spliced RNAs, differing only in that the splicing of its RNA is, in addition, growth temperature sensitive. We have generated N-nitroso-N-methylurea (NMU)-treated MuSVts110 revertants in which splicing was virtually complete at all temperatures and have investigated the molecular basis of this reversion on the assumption that the findings would reveal cis-acting elements controlling MuSVts110 splicing thermosensitivity. In a representative revertant (NMU-20), we found that complete splicing was conferred by a G-to-A substitution generating a consensus branchpoint (BP) signal (-CCCUGGC- to -CCCUGAC- [termed G(-25)A]) at -25 relative to the 3' ss. Weakening this BP to -CCCGAC- [G(-25)A,U(-27)C] moderately reduced splicing at the permissive temperature and sharply inhibited splicing at the originally nonpermissive temperature, arguing that MuSVts110 splicing thermosensitivity depends on a suboptimal BP-U2 small nuclear RNA interaction. This conclusion was supported by results indicating that lengthening the short MuSVts110 polypyrimidine tract and altering its uridine content doubled splicing efficiency at permissive temperatures and nearly abrogated splicing thermosensitivity. In vitro splicing experiments showed that MuSVts110 G(-25)A RNA intermediates were far more efficiently ligated than RNAs carrying the wild type BP, the G(-25)A,U (-27)C BP, or the extended polypyrimidine tract. The efficiency of ligation in vitro roughly paralleled splicing efficiency in vivo [G(-25)A BP > extended polypyrimidine tract > G(-25)A,U(-27)C BP > wild-type BP]. These results suggest that MuSVts110 RNA splicing is balanced by cis elements similar to those operating in other retroviruses and, in addition, that its splicing thermosensitivity is a response to the presence of multiple suboptimal splicing signals. PMID- 7494284 TI - Human papillomavirus type 16 variant lineages in United States populations characterized by nucleotide sequence analysis of the E6, L2, and L1 coding segments. AB - Human papillomavirus type 16 (HPV16) nucleotide sequence variations in the E6 (nucleotide positions [nt] 104 to 559), L2 (nt 4272 to 5657), and L1 (nt 5665 to 7148) open reading frames (ORFs), and the long control region (nt 7479 to 7842), were examined in 29 selected United States isolates. Of 3,690 nucleotide positions, 129 (3.5%) varied. The maximum pairwise distance was 66 nucleotide differences, or 1.8%. Nucleotide variations within different genome segments were phylogenetically compatible, and nucleotide changes within E6, L2, and L1 contained phylogenetic information beyond that provided in the long control region. Most isolates were classified as members of HPV16 lineages that have been described previously. However, two novel phylogenetic branches were identified. The L2 ORF was the most variable coding segment. L2 synonymous and nonsynonymous nucleotide changes were distributed asymmetrically. The amino-terminal half of the L2 protein was remarkably conserved among all isolates, suggesting that the region is under evolutionary constraint. The amino-terminal region of the E6 ORF was relatively varied, especially at E6 amino acid positions 10 and 14. Several amino acid difference in the L1 ORF were observed between lineages. Forty-nine amino acid variations across all sequenced coding regions were observed. These amino acid differences may be relevant to differences in the generation of humoral or cell-mediated immune responses to HPV16 variants. Our data form a basis for considering HPV16 sequence variation in the rational design of vaccine strategies and as an epidemiologic correlate of cervical cancer risk. PMID- 7494285 TI - A single hamster PrP amino acid blocks conversion to protease-resistant PrP in scrapie-infected mouse neuroblastoma cells. AB - Neurodegeneration caused by the transmissible spongiform encephalopathies is associated with the conversion of a normal host protein, PrP-sen, into an abnormal aggregated protease-resistant form, PrP-res. In scrapie-infected mouse neuroblastoma cells, mouse PrP-sen is converted into PrP-res but recombinant hamster PrP-sen expressed in these cells is not. In the present studies, recombinant hamster/mouse PrP-sen molecules were expressed in these scrapie infected cells to define specific PrP amino acid residues critical for the conversion to PrP-res. The results showed that homology to the region of mouse PrP-sen from amino acid residues 112 to 138 was required for conversion of recombinant PrP-sen to PrP-res in scrapie-infected mouse cells. Furthermore, a single hamster-specific PrP amino acid at residue 138 could inhibit the conversion of the recombinant PrP-sen into PrP-res. The data are consistent with studies in humans which show that specific amino acid residue changes within PrP can influence disease pathogenesis and transmission of transmissible spongiform encephalopathies across species barriers. PMID- 7494287 TI - Evolution of the Sindbis virus subgenomic mRNA promoter in cultured cells. AB - Transcription of the subgenomic mRNA of alphaviruses initiates at an internal site, called the promoter, which is highly conserved. To determine the functional significance of this conservation, we used an approach that randomizes positions 13 to -9 of the promoter to generate a library containing all possible sequences within this region, including the wild-type sequence. Viruses in the mixed population with more-efficient promoters were selected for during passaging in mammalian (BHK-21) cells. Results from early passage populations indicate that a large number of different promoters are functionally active. Analysis of eight individual viruses found that although each contained a promoter with different degrees of sequence identity to the wild-type sequence, all eight viruses produced progeny. This suggests that the mechanism for transcription allows for a diversity of sequences to serve as promoters. Further passaging of the viral library led to a population consensus sequence that increasingly resembled the wild-type sequence, despite the fact that these promoters are not constrained by the need to encode the carboxyl terminus of the nsP4 protein. Thus, conservation of the region of the promoter from -13 to -9 is in large part due to selection for promoter function, and the wild-type sequence and sequences closely similar to it seem to be optimal for promoter function in BHK-21 cells. PMID- 7494286 TI - Interaction of the 72-kilodalton human cytomegalovirus IE1 gene product with E2F1 coincides with E2F-dependent activation of dihydrofolate reductase transcription. AB - Three polypeptides are produced from the major immediate-early (IE) region of human cytomegalovirus by alternative splicing. The IE gene products regulate subsequent viral and cellular gene expression. We previously reported that cotransfection of a genomic clone of the major IE region stimulated transient expression of chloramphenicol acetyltransferase driven by the dihydrofolate reductase (DHFR) promoter and that an intact E2F site was required for the trans activation (M. Wade, T. F. Kowalik, M. Mudryj, E.-S. Huang, and J. C. Azizkhan, Mol. Cell. Biol. 12:4364-4374, 1992). With the availability of cDNA clones for the individual major IE proteins, we sought to determine which of these proteins exerted this effect and whether the IE protein(s) interacted with E2F. In this study, we use cotransfection to demonstrate that the 55- and 86-kDa major IE proteins from the IE2 region can each moderately trans activate the DHFR promoter and that the 72-kDa IE1 protein stimulates DHFR transcription to a much higher level. Furthermore, trans activation through the 72-kDa IE1 protein is in part E2F dependent, while activation by the 55- and 86-kDa IE proteins is E2F independent. We also demonstrate by in vitro pull-down assays that the 72-kDa IE1 protein can specifically interact with the DNA binding domain of E2F1 (amino acids 88 to 191) in the presence of nuclear extract. Moreover, antibodies to either E2F1 or IE72 will immunoprecipitate both E2F and IE72 from cells that stably express IE72, and antibody to E2F1 will immunoprecipitate IE72 from normal human fibroblast cells infected with human cytomegalovirus. PMID- 7494288 TI - Host-dependent evolution of the Sindbis virus promoter for subgenomic mRNA synthesis. AB - Alphaviruses are alternately transmitted between arthropod and vertebrate hosts. In each host, the virus transcribes a subgenomic mRNA that encodes the viral structural proteins which encapsidate the genome to form progeny virions. Transcription initiates at an internal site called the promoter. To determine if promoter utilization varies in mammalian versus mosquito cells, we used these cells as hosts to select for active promoters among a library of different mutant promoters. Compared with that in BHK-21 cells, selection was more rapid in mosquito (C7-10) cells, with much less diversity of promoters remaining after fewer passages. Thus, promoter selection is host dependent. With further passaging, both BHK-21 and C7-10 cells selected for similar sequences that closely resemble the wild-type promoter sequence. The difference in the rates of selection is not because BHK-21-derived promoters cannot function in mosquito cells. Instead, part of the host dependence is probably due to posttranscriptional differences between BHK-21 and C7-10 cells that may require more active promoters in mosquito cells. Part of the host dependence may also be attributed to the decreased rate of transcription versus that of replication in mosquito cells. This change in regulation of subgenomic to genomic RNA synthesis appears to correlate with the extent of cleavage or pausing of the genomic RNA synthesis at or close to the promoter. PMID- 7494289 TI - Genetic analysis of a hydrophobic domain of coxsackie B3 virus protein 2B: a moderate degree of hydrophobicity is required for a cis-acting function in viral RNA synthesis. AB - Coxsackie B virus protein 2B contains near its C terminus a hydrophobic domain with an amino acid composition that is characteristic for transmembrane regions. A molecular genetic approach was followed to define the role of this domain in virus reproduction and to study the structural and hydrophobic requirements of the domain. Nine substitution mutations were introduced in an infectious cDNA clone of coxsackie B3 virus. The effects of the mutations were studied in vivo by transfection of Buffalo green monkey cells with copy RNA transcripts. The results reported here suggest that a critical degree of hydrophobicity of the domain is essential for virus growth. The mutations S77M, C75M, I64S, and V66S, which caused either a small increase or decrease in mean hydrophobicity, yielded viable viruses. The double mutations S77M/C75M and I64S/V6-6S, which caused a more pronounced increase or decrease in hydrophobicity, were nonviable. Negatively charged residues (mutations A71E, I73E, and A71E/I73E) abolished virus growth. The mutations had no effect on the synthesis and processing of the viral polyprotein. Replication and complementation were studied by using a subgenomic coxsackievirus replicon containing the luciferase gene in place of the capsid coding region. Analysis of luciferase accumulation demonstrated that the mutations cause primary defects in viral RNA synthesis that cannot be complemented by wild-type protein 2B provided in trans. The hydrophobic domain is predicted by computer analysis to form a multimeric transmembrane helix. The proposed interaction with the membrane and the implications of the mutations on this interaction are discussed. PMID- 7494290 TI - Suppression of cellular proliferation by the papillomavirus E2 protein. AB - Carcinogenic progression of a human papillomavirus (HPV)-infected cell is often associated with integration of the viral genome in a manner which results in the loss of expression of the viral regulatory protein E2. One function of E2 is the regulation of expression of the viral oncogenes, E6 and E7. Introduction of the bovine papillomavirus type 1 (BPV-1) E2 transactivator (E2-TA) in HeLa cells, an HPV type 18 (HPV-18)-positive cervical carcinoma cell line results in growth arrest. In this study, we have found that the HPV-16 and HPV-18 E2 proteins share with BPV-1 E2-TA the ability to suppress HeLa cell growth. This property was not observed for the BPV-1 E2 transcriptional repressor (E2-TR). Analysis of various mutant E2 proteins for growth suppression revealed a requirement for the intact transactivation and DNA binding domains. A HeLa cell line (HeLa-tsE2) which expressed a conditional mutant E2 protein that was functional only at the permissive temperature (32 degrees C) was established, permitting an analysis of the molecular and cellular consequences of E2 expression. Our data indicate that one mechanism by which E2 suppresses cellular growth is through repression of E6 and E7 expression, thereby enabling the cellular targets of E6 and E7 to resume regulation of the cell cycle. PMID- 7494291 TI - Role of B cells and cytotoxic T lymphocytes in clearance of and immunity to rotavirus infection in mice. AB - The immune mechanisms involved in clearance of and immunity to rotavirus infection are poorly understood. Although mice with severe combined immunodeficiency (SCID mice) become chronically infected, nude mice have been reported to clear rotavirus infection similarly to immunocompetent controls. To better characterize the role of cytotoxic T lymphocytes (CTLs) in clearance of and immunity to rotavirus infection, we infected naive or previously infected beta 2-microglobulin (beta 2m) knockout mice with murine rotavirus. Naive beta 2m knockout mice shed rotavirus antigen 2 days longer than did normal control mice but completely resolved primary infection. beta 2m knockout naive mice treated with depleting doses of an anti-CD8 monoclonal antibody before infection shed viral antigen for an additional day. Upon rechallenge, beta 2m knockout mice, either treated with the anti-CD8 antibody or not treated, were completely resistant to reinfection. Clearance of rotavirus infection in naive beta 2m knockout mice correlated with the development of intestinal rotavirus-specific immunoglobulin A. Before rechallenge, beta 2m knockout mice had high levels of intestinal rotavirus-specific immunoglobulin A. These findings suggest that CTLs mediate rotavirus clearance but are not required for this function and that CTLs are not necessary for development of immunity to rotavirus reinfection. To further characterize the effector mechanisms involved in clearance and prevention of rotavirus infection, similar studies were performed with B-cell-deficient JHD knockout mice. After primary infection, most naive JHD mice had similar virus shedding clearance curves as did control mice and completely resolved primary infection. However, 2 of 29 became chronically infected. All JHD mice treated with anti-CD8 antibody became chronically infected with murine rotavirus. Upon rechallenge, JHD mice which had cleared primary infection were all susceptible to reinfection. These findings suggest that B cells also play a role in clearance of primary infection but are absolutely necessary for development of immunity against rotavirus reinfection. PMID- 7494292 TI - In vivo and in vitro association of hsc70 with polyomavirus capsid proteins. AB - Members of the 70-kDa family of cellular stress proteins assit in protein folding by preventing inappropriate intra- and intermolecular interactions during normal protein synthesis and transport and when cells are exposed to a variety of environmental stresses. During infection of A31 mouse fibroblasts with polyomavirus, the constitutive form of hsp70, hsc70, coimmunoprecipitated with all three viral capsid proteins (VP1, VP2, and VP3). In addition, the subcellular location of hsc70 changed from cytoplasmic to nuclear late in polyomavirus infection, coincident with the nuclear localization of the viral capsid proteins. VP1 and VP2 expressed in Sf9 insect cells with recombinant baculovirus vectors also coimmunoprecipitated with an hsp70-like protein, and VP1 expressed in Escherichia coli coimmunoprecipitated with the hsp70 homolog DnaK. Capsid proteins expressed by in vitro translation coimmunoprecipitated with the hsc70 protein present in the reticulocyte translation extract. Therefore, the polyomavirus capsid proteins associate with hsc70 during virus infection as well as in recombinant protein expression systems. This association may play a role in preventing the premature assembly of capsids in the cytosol and/or in facilitating the nuclear transport of capsid protein complexes. PMID- 7494293 TI - Downregulation of Lck-mediated signal transduction by tip of herpesvirus saimiri. AB - A protein, called tip, of herpesvirus saimiri associates with Lck in transformed T cells. To investigate the effects of complex formation on cellular signal transduction, we constructed human Jurkat-T-cell lines expressing tip. The expression of tip in Jurkat-T cells dramatically suppressed cellular tyrosine phosphorylation and surface expression of lymphocyte antigens. The expression of tip also blocked the induction of tyrosine phosphorylation by anti-CD3 stimulation. The expression of tip in fibroblast cells suppressed the transforming activity of oncogenic F505 Lck. Binding assays showed that the SH3 domain of Lck is sufficient to form a stable complex with tip in vitro. These results demonstrate that tip acts at an early stage of the T-cell signal transduction cascade by associating with Lck and downregulating Lck-mediated activation. Inhibition of Lck-mediated signal transduction by tip in T cells appears to be analogous to the inhibition of Lyn/Syk-mediated signal transduction in B cells by LMP2A of the B-cell-tropic Epstein-Barr virus. PMID- 7494294 TI - The dual-specificity phosphatase encoded by vaccinia virus, VH1, is essential for viral transcription in vivo and in vitro. AB - The genetic complexity of vaccinia virus is such that as well as encoding its own transcription and replication machinery, it encodes two protein kinases and a protein phosphatase. The latter enzyme, designated VH1, is a prototype for the dual-specificity class of phosphatases. Here we report that the H1 phosphatase is encapsidated within vaccinia virions and describe the construction of a viral recombinant in which expression of the H1 gene is regulated by the presence or absence of isopropylthiogalactopyranoside (IPTG) in the culture medium. When expression of H1 is repressed, the number of viral particles produced is not compromised but the fraction of these particles which is infectious is significantly reduced. The lack of infectivity of the H1-deficient particles is specifically correlated with their inability to direct the transcription of early genes either in vitro or in vivo. A proximal role for the viral phosphatase in regulating the onset of viral gene expression is implied. Prominent among the encapsidated proteins found to be hyperphosphorylated in H1-deficient virions is the 11-kDa product of the F18 gene; this protein is the major DNA-binding component of the viral nucleoprotein complex. The ability of recombinant H1 phosphatase to reverse this hyperphosphorylation in permeabilized virions strengthens the conclusion that the F18 protein is a bona fide substrate for the H1 phosphatase. PMID- 7494295 TI - The 3' untranslated region of picornavirus RNA: features required for efficient genome replication. AB - The role of the 3' untranslated region (3'UTR) in the replication of enteroviruses has been studied with a series of mutants derived from either poliovirus type 3 (PV3) or a PV3 replicon containing the reporter gene chloramphenicol acetyltransferase. Replication was observed when the PV3 3'UTR was replaced with that of either coxsackie B4 virus, human rhinovirus 14 (HRV14), bovine enterovirus, or hepatitis A virus, despite the lack of sequence and secondary structure homology of the 3'UTRs of these viruses. The levels of replication observed for recombinants containing the 3'UTRs of hepatitis A virus and bovine enterovirus were lower than those for PV3 and the other recombinants. Extensive site-directed mutagenesis of the single stem-loop structure formed by the HRV14 3'UTR indicated the importance of (i) the loop sequence, (ii) the stability of the stem, and (iii) the location of the stem immediately upstream of the poly(A) tail. The role of a 4-bp motif at the base of the HRV14 stem, highly conserved among rhinoviruses, was examined by site-directed mutagenesis of individual base pairs. This analysis did not pinpoint a particular base pair as crucial for function. The requirement for immediate adjacent positioning of the open reading frame and the 3'UTR was examined by insertion of a 1.1-kb heterologous sequence. A replicon containing this insert replicated to about 30% of the level observed for the wild type. However, the corresponding virus consistently deleted most of the inserted fragment, suggesting that its presence was incompatible with a full replication cycle. PMID- 7494296 TI - Plus-strand strong-stop DNA synthesis in retrotransposon Ty1. AB - Reverse transcription in the yeast retrotransposon Ty1 follows the general "rules" of retroviral replication overall. However, some details of the retroviral and Ty1 reverse transcription processes are different. We have identified and determined the structure of plus-strand strong-stop DNA and examined the effect of polypurine tract deletion mutations on its synthesis. Furthermore, we have defined the stop signal for plus-strand strong-stop DNA synthesis as an unusual 2'-O-ribosylated nucleotide in the primer tRNA. Full length plus-strand strong-stop DNA, following strand transfer, would have a terminal 2-base mismatch with minus-strand DNA. These findings indicate that the mechanism of plus-strand strong-stop DNA transfer in Ty1 differs from that of the retroviral transfer and suggest that full-length plus-strand strong-stop DNA is not a direct intermediate in Ty1 retrotransposition. PMID- 7494297 TI - Regulation of coronavirus mRNA transcription. AB - Coronaviruses synthesize a nested set of six to eight subgenomic (sg) mRNAs in infected cells. These mRNAs are produced in different, but constant, molar ratios. It is unclear which factors control the different levels of sg mRNAs. To determine whether the intergenic sequence (IS) involved in sg mRNA synthesis could affect the transcription efficiencies of other ISs and in this way regulate transcription levels, we inserted multiple ISs at different positions into a mouse hepatitis virus defective interfering RNA. Quantitation of the sg RNAs produced by identical ISs in different sequence contexts led to the following conclusions: (i) transcription efficiency depends on the location of the IS in the defective interfering virus genome, (ii) downstream ISs have a negative effect on transcription levels from upstream ISs, and (iii) upstream ISs have little or no effect on downstream ISs. The observation that a downstream IS downregulates the amounts of sg RNA produced by an upstream IS explains why the smaller sg RNAs are, in general, produced in larger quantities than the larger sg RNAs. Our data are consistent with coronavirus transcription models in which ISs attenuate transcription. In these models, larger sg RNAs are synthesized in smaller amounts because they encounter more attenuating ISs during their synthesis. PMID- 7494298 TI - Cellular factors required for papillomavirus DNA replication. AB - In vitro replication of papillomavirus DNA has been carried out with a combination of purified proteins and partially purified extracts made from human cells. DNA synthesis requires the viral E1 protein and the papillomavirus origin of replication. The E2 protein stimulates DNA synthesis in a binding site independent manner. Papillomavirus DNA replication is also dependent on the cellular factors replication protein A, replication factor C, and proliferating cell nuclear antigen as well as a phosphocellulose column fraction (IIA). Fraction IIA contains DNA polymerase alpha-primase and DNA polymerase delta. Both of these polymerases are essential for papillomavirus DNA replication in vitro. However, unlike the case with T-antigen-dependent replication from the simian virus 40 origin, purified DNA polymerase alpha-primase and delta cannot efficiently replace fraction IIA in the replication reaction. Hence, additional cellular factors seem to be required for papillomavirus DNA replication. Interestingly, replication factor C and proliferating-cell nuclear antigen are more stringently required for DNA synthesis in the papillomavirus system than in the simian virus 40 in vitro system. These distinctions indicate that there must be mechanistic differences between the DNA replication systems of papillomavirus and simian virus 40. PMID- 7494299 TI - A redundant nuclear protein binding site contributes to negative regulation of the mouse mammary tumor virus long terminal repeat. AB - The tissue specificity of mouse mammary tumor virus (MMTV) expression is controlled by regulatory elements in the MMTV long terminal repeat (LTR). These regulatory elements include the hormone response element, located approximately between -200 and -75, as well as binding sites for NF-1, Oct-1 (OTF-1), and mammary gland enhancer factors. Naturally occurring MMTV deletion variants isolated from T-cell and kidney tumors, transgenic-mouse experiments with MMTV LTR deletions, and transient transfection assays with LTR constructs indicate that there are additional transcription regulatory elements, including a negative regulatory element (NRE), located upstream of the hormone response element. To further define this regulatory region, we have constructed a series of BAL 31 deletion mutants in the MMTV LTR for use in transient transfection assays. These assays indicated that deletion of two regions (referred to as promoter-distal and -proximal NREs) between -637 and -201 elevated basal MMTV promoter activity in the absence of glucocorticoids. The region between -637 and -264 was surveyed for the presence of nuclear protein binding sites by gel retardation assays. Only one type of protein complex (referred to as NRE-binding protein or NBP) bound exclusively to sites that mapped to the promoter-distal and -proximal NREs identified by BAL 31 mutations. The promoter-proximal binding site was mapped further by linker substitution mutations and transfection assays. Mutations that mapped to a region containing an inverted repeat beginning at -287 relative to the start of transcription elevated basal expression of a reporter gene driven by the MMTV LTR. A 59-bp DNA fragment from the distal NRE also bound the NBP complex. Gel retardation assays showed that mutations within both inverted repeats of the proximal NRE eliminated NBP binding and mutations within single repeats altered NBP binding. Intriguingly, the NBP complex was detected in extracts from T cells and lung cells but was absent from mammary gland cells. These results suggest that a factor contributing to high-level expression of MMTV in the mammary gland is the lack of negative regulation by NBP. PMID- 7494300 TI - Distribution of baboon endogenous virus among species of African monkeys suggests multiple ancient cross-species transmissions in shared habitats. AB - PCR amplification of baboon endogenous virus (BaEV) long terminal repeat, reverse transcriptase gene, and env fragments from 24 different species of African monkeys indicates that BaEV is less widespread than was formerly thought. Instead of being present in every species of African primates, BaEV can be found only in baboons, geladas, and mangabeys (all belonging to the Papionini tribe) and in African green monkey (Cercopithecus aethiops)subspecies. BaEV, which can be activated from baboon and gelada tissues, was most likely introduced in the germ line only recently (less than a few million years ago) and has not been inherited from a common ancestor of all extant African monkeys. Neighbor-joining and maximum-likelihood analyses of the sequences obtained showed that two distinct virus clusters can be distinguished: the first containing baboon, gelada, and African green monkey BaEV sequences and the second consisting of mandrill and mangabey BaEV sequences. This viral evolutionary tree does not follow host phylogeny, indicating the cross-species transmissions and multiple germ line fixations of the virus must have occurred in the past. BaEV sequences are found in monkeys inhabiting savannas (baboons, geladas, and African green monkeys) as well as forests (managabeys and mandrills) and cluster according to the habitats of their hosts, providing evidence for cross-species transmission in shared habitats. PMID- 7494301 TI - Retention of oncogenicity by a Marek's disease virus mutant lacking six unique short region genes. AB - We previously reported the construction of Marek's disease virus (MDV) strains having mutations in various genes that map to the unique short (US) region of the viral genome (J.L. Cantello, A.S. Anderson, A. Francesconi, and R.W. Morgan, J. Virol. 65:1584-1588, 1991; M.S. Parcells, A.S. Anderson, and R.W. Morgan, Virus Genes 9:5-13, 1994; M.S. Parcells, A.S. Anderson, and R.W. Morgan, J. Virol. 68:8239-8253, 1994). These strains were constructed by using a high-passage-level serotype 1 MDV strain which grew well in chicken embryo fibroblasts. Despite the growth of the parent and mutant viruses in cell culture, in vivo studies were limited by poor growth of these strains in chickens. One of the mutants studied lacked 4.5 kbp of US region DNA and contained the lacZ gene of Escherichia coli inserted at the site of the deletion. The deletion removed MDV homologs to the US1, US2, and US10 genes of herpes simplex virus type 1 as well as three MDV specific open reading frames. We now report the construction of a mutant MDV containing a similar deletion in the US region of the highly oncogenic RB1B strain. This mutant, RB1B delta 4.5lac, had a growth impairment in established chicken embryo fibroblasts similar to that described previously for MDVs lacking a functional US1 gene. In chickens, RB1B delta 4.5lac showed decreased early cytolytic infection, mortality, tumor incidence, and horizontal transmission. Several lymphoblastoid cell lines were established from RB1B delta 4.5lac-induced tumors, and virus reactivated from these cell lines was LacZ+. These results indicate that the deleted genes are nonessential for the transformation of chicken T cells or for the establishment and maintenance of latency. On the basis of the growth impairment observed for RB1B delta 4.5lac in cell culture and in vivo, we conclude that deletion of these genes affects the lytic replication of MDV. This is the first MDV mutant constructed in the RB1B oncogenic strain, and the methodology described herein provides for the direct examination of MDV encoded determinants of oncogenicity. PMID- 7494302 TI - Two herpes simplex virus type 1 latency-active promoters differ in their contributions to latency-associated transcript expression during lytic and latent infections. AB - Herpes simplex virus type 1 (HSV-1) establishes latency in human sensory ganglia, during which time the viral genome is transcriptionally silent with the exception of the latency-associated transcripts (LATs). The most abundant LAT is a 2-kb RNA whose biosynthesis is poorly characterized. The 2-kb LAT may be a primary transcript, or its synthesis may involve splicing and/or other forms of processing. Two potential RNA polymerase II promoters (LAP1 and LAP2) upstream of the 2-kb LAT 5' end have been identified. To investigate the role played by LAP1 and LAP2 in the synthesis of the 2-kb LAT under lytic and latent conditions, we analyzed HSV-1 mutants which contain deletions of one or both of these promoters. During lytic infection in cell culture, the cis elements critical for the normal accumulation of the 2-kb LAT were mapped to LAP2, while LAP1 sequences were largely dispensable. The 5' ends of the major 2-kb LATs produced by the wild-type and LAP deletion viruses were examined by primer extension analysis and were all found to be identical (+/- 2 bp). The accumulation of the 2-kb LAT during latent infections of murine trigeminal ganglia was examined by Northern (RNA) blot and by reverse transcription-PCR. In contrast to the results found in lytic infections, the critical cis elements needed for 2-kb LAT accumulation during latency were mapped to LAP1. Deletion of LAP1 resulted in a 500-fold reduction in 2-kb LAT accumulation, whereas deletion of LAP2 resulted in only a 2- to 3-fold reduction. Deletion of both LAP1 and LAP2 resulted in undetectable levels of the 2-kb LAT. Our results indicate that both LAP1 and LAP2 are critical for 2-kb LAT expression but under different conditions. LAP1 is essential for LAT expression during latency, while LAP2 is primarily responsible for LAT expression in lytic infections in cell culture. LAP1 and LAP2 may prove to be functionally independent promoter elements that control 2-kb LAT expression during different stages of HSV-1 infections. PMID- 7494303 TI - Mutational analysis of cell cycle arrest, nuclear localization and virion packaging of human immunodeficiency virus type 1 Vpr. AB - Human immunodeficiency virus type 1 Vpr is a virion-associated, regulatory protein that is required for efficient viral replication in monocytes/macrophages. The protein is believed to act in conjunction with the Gag matrix protein to allow import of the viral preintegration complex in nondividing cells. In cells, Vpr localizes to the nucleus. Recently, we showed that Vpr prevents the activation of p34cdc2-cyclin B. This results in arrest of Vpr expressing cells in the G2/M phase of the cell cycle. Here, we use a panel of expression vectors encoding Vpr molecules mutated in the amino-terminal alpha helical region, the central hydrophobic region, or the carboxy-terminal basic region to define the functional domains of the protein. The results showed cell cycle arrest was largely controlled by the carboxy-terminal basic domain of the protein. In contrast, the amino-terminal alpha-helical region of Vpr was required for nuclear localization and packaging into virions. The carboxy terminus appeared to be unnecessary for nuclear localization. In the alpha-helical region, mutation of Ala-30 to Pro resulted in a protein that localized to the cytoplasm. Surprisingly, fusion of Vpr to luciferase resulted in a molecule that failed to localize to the nucleus. In addition, we show that simian immunodeficiency virus Vpr, but not Vpx, induces G2 arrest. We speculate that Vpr has two sites for interaction with cellular factors: one in the alpha-helical region that specifies nuclear localization and one in the carboxy-terminal domain that is required for Cdc2 inhibition. PMID- 7494304 TI - E1A promotes association between p300 and pRB in multimeric complexes required for normal biological activity. AB - The oncogenes of the small DNA tumor viruses encode transforming proteins with multiple domains that influence the cell cycle and aspects of the transformed phenotype. Like other gene products of this type, the adenovirus E1A proteins influence the cell by binding to specific cell growth control proteins. These include members of the retinoblastoma gene product (pRB) family, which are bound by the E1A region 2-specific site, and p300, which is bound at the E1A amino terminus. Binding at these two sites is largely independent, and discrete transcription-regulating functions remain intact in E1A products when only one or the other binding site is functional. In this report, immunoprecipitation with p300 antibodies reveals the presence of the pRB family proteins in p300 complexes when E1A is expressed in host cells, indicating that E1A can mediate physical contact between p300 and the pRB-related proteins. The ability of E1A to induce proliferation efficiently in quiescent primary cells correlates closely with the ability to bind p300 and individual members of the pRB family simultaneously in multimeric complexes, even though the E1A active sites can bind their target proteins efficiently when separated on different molecules. Conservation of a spacer region between the two binding sites that is required for simultaneous binding and efficient induction of proliferation supports the concept that the E1A protein structure has evolved to facilitate simultaneous binding. These results indicate that the E1A proteins are designed not merely to sequester these cellular products, but also to bring them into proximal association with each other in biologically significant complexes. PMID- 7494305 TI - Genetic and structural analysis of a virulence determinant in polyomavirus VP1. AB - The LID strain of polyomavirus differs from other laboratory strains in causing a rapidly lethal infection of newborn C3H/Bi mice. This virulent behavior of LID was attenuated by dilution, yet at sublethal doses LID was able to induce tumors at a high frequency, like its parent virus PTA. By constructing and assaying LID PTA recombinant viruses and by DNA sequencing, the determinant of virulence in LID was mapped to the major viral capsid protein, VP1. The VP1s of LID and PTA differed at two positions: at 185, LID has phenylalanine and PTA has tyrosine, and at 296, LID has alanine and PTA has valine. Results obtained with viruses constructed by site-directed mutagenesis showed that alanine at position 296 is sufficient to confer a fully virulent phenotype regardless of which amino acid is at position 185. However, with valine at position 296, an effect of phenylalanine at position 185 is apparent, as this virus possesses an intermediate level of virulence. A crystal structure of polyomavirus complexed with 3'-sialyl lactose previously indicated van der Waals contacts between the side chain of valine 296 and the sialic acid ring (T. Stehle, Y. Yan, T. L. Benjamin, and S. C. Harrison, Nature [London] 369:160-163, 1994). When this interaction was modeled with alanine, these contacts were greatly reduced. Direct confirmation that the substitutions in VP1 affected receptor binding was obtained by studying virus hemagglutination behavior. The ensemble of results are discussed in terms of the idea that a lower affinity of the virus for its receptor can result in more rapid spread and increased pathogenicity. PMID- 7494306 TI - VP16 interacts via its activation domain with VP22, a tegument protein of herpes simplex virus, and is relocated to a novel macromolecular assembly in coexpressing cells. AB - In addition to its function as a powerful transactivator of viral immediate-early transcription, VP16 is an essential component of the herpes simplex virus (HSV) virion. As such, VP16 is introduced into cells, to effect its function in transactivation, as part of the virus tegument. Here we examine the potential for VP16 protein-protein interactions specific to virus-infected cells and show that VP16 copurifies in a highly enriched fraction with a single major polypeptide which we identify as the virus-encoded structural protein VP22. We further show that in vitro-translated VP22 binds specifically to purified VP16. The activation domain of VP16 was required and largely sufficient for this binding. Mutations within this domain, which disrupt its transactivation function, also affected VP22 binding. Furthermore, we show that while VP16 and VP22 showed distinct patterns of compartmentalization in vivo, coexpression of both proteins resulted in a profound reorganization from their normal locations to a novel macromolecular assembly. The colocalization was also dependent on the activation domain of VP16 but required additional determinants within the N terminus. These results are discussed in the context of VP16 regulation of transcription both early in infection during delivery of tegument proteins and at late times during virus assembly. PMID- 7494307 TI - Neurons differentially control expression of a herpes simplex virus type 1 immediate-early promoter in transgenic mice. AB - The immediate-early proteins of herpes simplex virus control the cascade of viral gene expression during lytic infection. It is not known which viral or host proteins control the reactivation of the viral genome in latently infected neurons. To determine whether neuronal proteins can regulate a herpes simplex virus immediate-early promoter in vivo, transgenic mice containing the promoter regulatory region of the herpes simplex virus type 1 immediate-early gene (ICP4) fused to the bacterial beta-galactosidase gene were generated. Two lines of mice, in the absence of viral proteins, displayed ICP4 promoter activity in neurons in specific locations in the central nervous system. The anatomic locations of these neurons were the hippocampus, cerebellar cortex, superior colliculus, indusium griseum, mammillary nucleus, cerebral cortex, and the dorsal laminae of the dorsal horns of the spinal cord. Additional subsets of neurons expressed the ICP4 promoter at lower levels; these included trigeminal ganglia and retinas. In a third line of mice, lower levels of expression were present in many of the above described neurons. Many types of neurons, nearly all nonneuronal cells in the central nervous system, and some non-nervous system tissues were negative. Viral proteins including VP16 are not necessary to induce transcription from the ICP4 promoter in many neurons and some other cell types but may be required in most cells in vivo. An approximately 100-fold-greater number of neurons in the trigeminal ganglia expressed ICP4 promoter activity in newborn mice compared with adults. These data provide direct evidence that host proteins are sufficient to activate a herpes simplex virus immediate-early promoter in neurons in vivo and that a differential expression pattern for this promoter exists within different neuronal phenotypes and between the same neurons in different ages of mice. PMID- 7494308 TI - Envelopment of varicella-zoster virus: targeting of viral glycoproteins to the trans-Golgi network. AB - Previous studies suggested that varicella-zoster virus derives its final envelope from the trans-Golgi network (TGN) and that envelope glycoproteins (gps) are transported to the TGN independently of nucleocapsids. We tested the hypothesis that gpI is targeted to the TGN as a result of a signal sequence or patch encoded in its cytosolic domain. cDNAs encoding gpI wild type (wt) and a truncated mutant gpI(trc) lacking transmembrane and cytosolic domains were cloned by using the PCR. Cells transfected with cDNA encoding gpI(wt) or gpI(trc) synthesized and N glycosylated the proteins. gpI(wt) accumulated in the TGN, some reached the plasmalemma, but none was secreted. In contrast, gpI(trc) was retained and probably degraded in the endoplasmic reticulum; none was found on cell surfaces, but some was secreted. The distribution of gpI(trc) was not affected by deletion of potential glycosylation sites. To locate a potential gpI-targeting sequence, cells were transfected with cDNA encoding chimeric proteins in which the ectodomain of a plasmalemmal marker, the interleukin-2 receptor (tac), was fused to different domains of gpI. A chimeric protein in which tac was fused with the transmembrane and cytoplasmic domains of gpI was targeted to the TGN. In contrast, a chimeric protein in which tac was fused only with the gpI transmembrane domain passed through the TGN and concentrated in endosomes. We conclude that gpI is targeted to the TGN as a result of a targeting sequence or patch in its cytosolic domain. PMID- 7494309 TI - Human cytomegalovirus IE1 and IE2 proteins block apoptosis. AB - Human cytomegalovirus-infected fibroblasts are resistant to the induction of apoptosis by superinfection with a mutant adenovirus unable to produce the viral E1B 19-kDa protein that normally causes an E1A protein-mediated apoptotic response. Two cytomegalovirus gene products that block apoptosis were identified. The IE1 and IE2 proteins each inhibit the induction of apoptosis by tumor necrosis factor alpha or by the E1B 19-kDa-protein-deficient adenovirus but not by irradiation with UV light. Our results suggest a new physiological role for the IE1 and IE2 proteins in the human cytomegalovirus replication cycle. PMID- 7494311 TI - Genotypic selection of simian immunodeficiency virus in macaque infants infected transplacentally. AB - To understand viral and host factors that contribute to transplacental transmission of human immunodeficiency virus, we developed an animal model using pregnant female macaques infected with simian immunodeficiency virus (SIV). Pregnant females were inoculated intravenously during midgestation with either a well-characterized primary isolate of SIV (SIV/DeltaB670) or a combination of SIV/DeltaB670 and the macrophage-tropic molecular clone SIV/17E-Fr. The viral genetic diversity in five infected female macaques and their in utero-infected infants was analyzed. All of the mothers harbored a genetically diverse virus population at parturition, whereas a single genotype from the maternal quasispecies was identified in the infants at birth. Only one of two variants was found in the infants: SIV/17E-Fr (two cases) or a genotype contained within the SIV/DeltaB670 quasispecies (three cases). The macrophage-tropic properties of both transmitted genotypes were suggested by productive replication in primary rhesus macrophage cultures in vitro and the clonal presence in central nervous system tissue of infected monkeys with encephalitis. These observations provide compelling evidence for both genotypic and phenotypic selection in transplacental transmission of SIV and suggest a critical role for macrophages in fetal infection in utero. PMID- 7494310 TI - Heteroduplex mobility assay and phylogenetic analysis of V3 region sequences of human immunodeficiency virus type 1 isolates from Gulu, northern Uganda. The Italian-Ugandan Cooperation AIDS Program. AB - We analyzed peripheral blood mononuclear cells from 19 asymptomatic seropositive pregnant women from the district of Gulu in northern Uganda. A 700-bp fragment of the human immunodeficiency virus type 1 (HIV-1) env gene, including the V3-V5 region, was successfully amplified by PCR from 10 samples (52.6%) and was subsequently subjected to both a heteroduplex mobility assay for genetic screening and subtyping and DNA sequence analysis (approximately 300 bp) for nucleotide comparison and phylogenetic studies. The results show the presence of HIV-1 A and D subtypes (or clades) in this rural area, with the prevalence of the A subtype (8 of 10) being greater than that of the D (2 of 10) subtype, which is unlike what was previously reported for Uganda. By pairwise comparison analysis, the percentage of sequence divergence among samples within each subtype is low (the average intrasubtype divergence is 15.8%), but it is significantly higher between the two subtypes (the average intersubtype divergence is 23%). At the amino acid level, the two HIV-1 subtypes show distinct tetramers at the apex of the V3 loop and, in particular, GPGQ in clade A and GPGR in clade D. In addition, 10 of the 19 viral samples (52.6%) have been isolated in vitro. Nine of the samples have been classified as rapid/high, which reflects a high in vitro replication capacity for the HIV-1 field isolates from this country, even for those obtained from seropositive asymptomatic individuals. These observations, despite being made on the basis of a limited sample size, show a modest degree of genetic divergence among samples isolated in the last 4 years in this country by comparison with those based on the 1990 data on HIV-1 isolates from Kampala. The results reported here are, therefore, extremely relevant for Uganda, which is one of the selected World Health Organization field sites for future HIV-1 vaccine evaluation programs. PMID- 7494312 TI - Genetic rearrangements occurring during a single cycle of murine leukemia virus vector replication: characterization and implications. AB - Retroviruses evolve at rapid rates, which is presumably advantageous for responding to selective pressures. Understanding the basic mutational processes involved during retroviral replication is important for comprehending the ability of retroviruses to escape immunosurveillance and antiviral drug treatment. Moreover, since retroviral vectors are important vehicles for somatic cell gene therapy, knowledge of the mechanism of retroviral variation is critical for anticipating untoward mutational events occurring during retrovirus-medicated gene transfer. The focus of this report is to examine the spectrum of genomic rearrangements arising during a single cycle of Moloney murine leukemia virus (MoMLV) vector virus replication. An MoMLV vector containing the herpes simplex virus thymidine kinase (tk) gene was constructed. MoMLV vector virus was produced in packaging lines, and target cells were infected. From a total of 224 mutant proviruses analyzed, 114 had gross rearrangements readily detectable by Southern blotting. The remaining proviruses were of parental size. PCR and DNA sequence analysis of 73 of the grossly rearranged mutant proviruses indicated they resulted from deletions, combined with insertions, duplications, and complex mutations that were a result of multiple genomic alterations in the same provirus. Complex hypermutations distinct from those previously described for spleen necrosis virus and human immunodeficiency virus were detected. There was a correlation between the mutation breakpoints and single-stranded regions in the predicted viral RNA secondary structure. The results also confirmed that the tk gene is inactivated at an average rate of about 8.8% per cycle of retroviral replication, which corresponds to a rate of mutation of 3%/kbp. PMID- 7494313 TI - Inducible expression of the P, V, and NP genes of the paramyxovirus simian virus 5 in cell lines and an examination of NP-P and NP-V interactions. AB - The P, V, and NP genes of the paramyxovirus simian virus 5 (SV5) were cloned such that their expression was regulated by the tetracycline-controlled transactivator (M. Gossen and H. Bujard, Proc. Natl. Acad. Sci. USA 89:5547-5551, 1992), and mammalian cell lines that inducibly expressed individually the P, V, or NP protein or coexpressed the P plus NP or V plus NP proteins were isolated. A plasmid that expresses the tetracycline-controlled transactivator linked, via the foot-and-mouth disease virus 2A cleavage peptide sequence, to the neomycin aminoglycoside phosphotransferase gene was constructed. Cells were cotransfected with this plasmid, and the appropriate responder plasmids and clonies were selected on the basis of their resistance to Geneticin (via the neomycin aminoglycoside phosphotransferase gene). The properties of these cell lines, in terms of the induction of the P, V, and NP genes, are described in detail. Both the P and V proteins were phosphorylated when expressed alone. In immunoprecipitation studies using a monoclonal antibody that recognizes both the P and V proteins, a nonphosphorylated host cell protein with an estimated molecular weight of 150,000 was coprecipitated with V but not P. Immunofluorescence data demonstrated that when expressed separately, the P protein had a diffuse cytoplasmic distribution, but the related V protein had both a nuclear and cytoplasmic distribution. The NP protein had a granular cytoplasmic distribution, giving rise to punctate and granular fluorescence. Coexpression of the NP and P proteins resulted in the accumulation of large cytoplasmic inclusion aggregates, similar to those visualized at late times in SV5-infected cells. Coexpression of V with NP led to a partial redistribution of the NP protein in that the NP protein had both a diffuse cytoplasmic and nuclear distribution in the presence of V, but no NP-V aggregates or inclusion bodies were visualized. Direct binding studies also revealed that NP bound to both P and V. For SV5, these studies suggest that V may have a role in keeping NP soluble prior to encapsidation. PMID- 7494314 TI - 5' Coding and regulatory region sequence divergence with conserved function of the Epstein-Barr virus LMP2A homolog in herpesvirus papio. AB - B-lymphotropic herpesviruses naturally infecting Old World primates share biologic, epidemiologic, pathogenic, and molecular features with the human pathogen Epstein-Barr virus (EBV). These related gammaherpesviruses have colinear genomes with considerable nucleotide homology. The replicative cycle genes share a high degree of homology across species, whereas the transformation-associated EBV latent genes appear to be much more divergent. For example, the EBV BamHI Nhet fragment, which encodes all or part of the EBV latent infection membrane proteins, cross-hybridizes poorly to DNA from nonhuman primate B-lymphotropic herpesviruses. A viral DNA fragment corresponding to this region of the EBV genome was isolated from the baboon B-lymphotropic herpesvirus, herpesvirus papio, and used to clone a herpesvirus papio cDNA corresponding to EBV LMP2A. At least three tyrosine kinase interaction motifs are conserved despite significant amino acid divergence of the herpesvirus papio LMP2A first exon from the EBV homolog. Functionally, the herpesvirus papio LMP2A is tyrosine phosphorylated and induces tyrosine phosphorylation of cell proteins similar to EBV LMP2A. The 12 hydrophobic LMP2 transmembrane domains are well conserved. Two CBP (Jk) binding sites important for EBNA-2-induced transactivation of the LMP2A promoter are also present in the herpesvirus papio LMP2A promoter, and the simian LMP2A promoter is also responsive to EBV EBNA-2-induced transactivation in human B cells. Thus, transcriptional regulation, splicing, kinase interaction sites, and tyrosine phosphorylation of the LMP2A homologs have been conserved despite significant sequences heterogeneity in the preterminal repeat regions of these human and nonhuman primate EBVs. The conservation of the LMP2 gene, despite its apparent nonessential role for in vitro EBV infection, suggests an important role for LMP2A in vivo. The similarities between these human and simian B-lymphotropic herpesviruses, and the LMP2 genes in particular, suggest that the function of LMP2 in vivo could be addressed by using recombinant LMP2A-mutant simian viruses and experimental infection of Old World primates. PMID- 7494315 TI - Human immunodeficiency virus type 1 envelope protein does not stimulate either prostaglandin formation or the expression of prostaglandin H synthase in THP-1 human monocytes/macrophages. AB - Prostaglandin E2 is observed at elevated levels during human immunodeficiency virus (HIV) infection and thus may contribute to the HIV-dependent immunosuppression. The mechanisms responsible for this increase are not understood. Evidence indicates that the viral envelope proteins perturb membrane signaling mediated by the CD4 receptor, suggesting that the free envelope protein and/or the intact virus may be responsible for the increase in prostaglandin E2 levels. In this study, we have used THP-1 human monocytes and THP-1 cells differentiated by 12-O-tetradecanoylphorbol-13-acetate treatment into macrophages to determine if the HIV envelope protein, gp120, or an anti-CD4 receptor antibody stimulates prostaglandin formation by interacting with the CD4 receptor. Incubation of THP-1 cells with OKT4A antibody greatly stimulated the CD4-p56lck receptor complex as estimated by enhanced p56lck autophosphorylation, while the gp120 gave small but significant responses. Monocytic THP-1 cells poorly metabolized arachidonic acid to prostaglandin E2 and thromboxane B2 as measured by high-pressure liquid chromatography analysis. Western blot (immunoblot) and Northern (RNA) blot analyses revealed that unstimulated monocytes expressed little prostaglandin H synthase 1 and 2 (PGHS-1 and -2). Incubation of the monocytes with lipopolysaccharide, OKT4A, or gp120 did not increase the formation of prostaglandins. The expression of PGHS-1 or PGHS-2 was also not increased. Differentiation of the monocytes to macrophages by 12-O-tetradecanoylphorbol-13 acetate treatment resulted in increased expression of PGHS-1 and increased formation of prostaglandins compared with that for the monocytes. Lipopolysaccharide stimulation of the macrophages increased the formation of prostaglandins and increased the expression of PGHS-2 in the macrophages. However, OKT4A or gp120 preparation, at concentrations that stimulated p56lck autophosphorylation, did not enhance the formation of prostaglandins or the expression of PGHS-1 or PGHS-2. OKT4A and gp120 also did not stimulate the release of arachidonic acid, indicating that phospholipase A2 was not activated by the CD4 receptor in either the THP-1 monocytes or macrophages. These results indicate that activation of the CD4-p56lck receptor signal transduction pathway by the HIV envelope protein does not increase prostaglandin formation. PMID- 7494316 TI - In vitro selection of RNA ligands for the ribosomal L22 protein associated with Epstein-Barr virus-expressed RNA by using randomized and cDNA-derived RNA libraries. AB - The Epstein-Barr virus (EBV)-expressed RNA 1 (EBER1) associates tightly with the ribosomal protein L22. We determined the general requirements for an RNA to bind L22 in a SELEX experiment, selecting RNA ligands for L22 from a randomized pool of RNA sequences by using an L22-glutathione S-transferase fusion protein. The selected sequences all contained a stem-loop motif similar to that of the region of EBER1 previously shown to interact with L22. The nucleotides were highly conserved at three positions within the stem-loop and identical to the corresponding nucleotides in EBER1. Two independent binding sites for L22 could be identified in EBER1, and mobility shift assays indicated that two L22 molecules can interact with EBER1 simultaneously. To search for a cellular L22 ligand, we constructed a SELEX library from cDNA fragments derived from RNA that was coimmunoprecipitated with L22 from an EBV-negative whole-cell lysate. After four rounds of selection and amplification, most of the clones that were obtained overlapped a sequence corresponding to the stem-loop between nucleotides 302 and 317 in human 28S ribosomal RNA. This stem-loop fulfills the criteria for optimal binding to L22 that were defined by SELEX, suggesting that human 28S ribosomal RNA is likely to be a cellular L22 ligand. Additional L22 binding sites were found in 28S ribosomal RNA, as well as within 18S ribosomal RNA and in RNA segments not present in sequence databases. The methodology described for the conversion of a preselected cellular RNA pool into a SELEX library might be generally applicable to other proteins for the identification of cellular RNA ligands. PMID- 7494318 TI - Identification of human telomeric repeat motifs at the genome termini of human herpesvirus 7: structural analysis and heterogeneity. AB - Human herpesvirus 6 (HHV-6) and HHV-7 are closely related T-lymphotropic betaherpesviruses which share a common genomic organization and are composed of a single unique component (U) that is bounded by direct repeats (DRL and DRR). In HHV-6, a sequences have been identified at each end of the DR motifs, resulting in the arrangement aDRLa-U-aDRRa. In order to determine whether determine whether HHV-7 contains similar a sequences, we have sequenced the DRL-U and U-DRR junctions of HHV-7 strain JI, together with the DRR.DRL junction from the head-to tail concatamer that is generated during productive virus infection. In addition, we have sequenced the genomic termini of an independent isolate of HHV-7. As in HHV-6, a (GGGTTA)n motif identical to the human telomeric repeat sequence (TRS) was identified adjacent to, but not at, the genome termini of HHV-7. The left genome terminus and the U-DRR junction contained a homolog of the consensus herpesvirus packaging signal, pac-1, followed by short tandem arrays of TRSs separated by single copies of a second 6-bp repeat. This organization is similar to the arrangement found at U-DRR in HHV-6 but differs from it in that the TRS arrays are considerably shorter in HHV-7. The right genome terminus and the DRL-U junction contained a homolog of the consensus herpesvirus packaging signal, pac 2, followed by longer tandem arrays of TRSs separated by single copies of either a 6-bp or a 14-bp repeat. This arrangement is considerably more complex than the simple tandem array of TRSs that is present at the corresponding genomic location in HHV-6 and corresponds to a site of both inter- and intrastrain heterogeneity in HHV-7. The presence of TRSs in lymphotropic herpesviruses from humans (HHV-6 and HHV-7), horse (equine herpesvirus 2), and birds (Marek's disease virus) is striking and suggests that these sequences may have functional or structural significance. PMID- 7494317 TI - The coxsackievirus A9 RGD motif is not essential for virus viability. AB - An RGD (arginine-glycine-aspartic acid) motif in coxsackievirus A9 has been implicated in internalization through an interaction with the integrin alpha v beta 3. We have produced a number of virus mutants, lacking the motif, which have a small-plaque phenotype in LLC-Mk2 and A-Vero cells and are phenotypically normal in RD cells. Substitution of flanking amino acids also affected plaque size. The results suggest that interaction between the RGD motif and alpha v beta 3 is not critical for virus viability in the cell lines tested and therefore that alternative regions of the CAV-9 capsid are involved in internalization. PMID- 7494319 TI - Preferential usage of the T-cell receptor by influenza virus hemagglutinin specific human CD4+ T lymphocytes: in vitro life span of clonotypic T cells. AB - Human helper T-cell (Th) responses to influenza A virus were studied by analyzing T-cell receptor V beta gene diversity in hemagglutinin-specific Th lymphocytes. The T-lymphocyte population from peripheral blood became quickly oligoclonal when stimulated in vitro with the HA306-329 peptide, and T-cell receptor V beta 3 genes were mainly expanded. Moreover, specific junctional region oligonucleotide probes corresponding to hemagglutinin-specific clones were used to estimate temporal diversity during antigenic stimulations. PMID- 7494320 TI - Effect of human papillomavirus type 16 oncogenes on MAP kinase activity. AB - The mitogen-activated protein (MAP) kinase signal transduction pathway is an intracellular signaling cascade which mediates cellular responses to growth and differentiation factors. The MAP kinase pathway can be activated by a wide range of stimuli dependent on the cell types, and this is normally a transient response. Oncogenes such as ras, src, raf, and mos have been proposed to transform cells in part by prolonging the activated stage of components within this signaling pathway. The human papillomavirus (HPV) oncogenes E6 and E7 play an essential role in the in vitro transformation of primary human keratinocytes and rodent cells. The HPV type 16 E5 gene has also been shown to have weak transforming activity and may enhance the epidermal growth factor (EGF)-mediated signal transduction to the nucleus. In the present study, we have investigated the effects of the oncogenic HPV type 16 E5, E6, and E7 genes on the induction of the MAP kinase signaling pathway. The E5 gene induced an increase in the MAP kinase activity both in the absence and in the presence of EGF. In comparison, the E6 and E7 oncoproteins do not alter the MAP kinase activity or prolong the MAP kinase activity induced with EGF. These findings suggest that E5 may function, at least in part, to enhance the cell response through the MAP kinase pathway. However, the transforming activity of E6 and E7 is not associated with alterations in the MAP kinase pathway. These findings are consistent with E5 enhancing the response to growth factor stimulation. PMID- 7494321 TI - Binding motifs predict major histocompatibility complex class II-restricted epitopes in the Sendai virus M protein. AB - Major histocompatibility complex (MHC) class I ligand motifs have been defined for a number of class I molecules and have been successfully used to identify class I-restricted cytotoxic T-cell epitopes. In contrast, the relative degeneracy of sequence motifs in naturally processed MHC class II ligands has suggested that they may be of more limited use. Here, we use a predicted I-Ab ligand motif to identify antigenic peptides in the Sendai virus Enders strain matrix (M) protein. The entire coding sequence of the M protein was derived, and seven peptide sequences that contained the predicted I-Ab motif were identified. Analysis of I-Ab-restricted M-specific T-cell hybridomas for reactivity to these synthetic peptides identified two distinct epitopes. These data demonstrate that MHC class II motifs can be valuable in predicting T-cell epitopes. PMID- 7494322 TI - Efficient nuclear localization and immortalizing ability, two functions dependent on the adenovirus type 5 (Ad5) E1A second exon, are necessary for cotransformation with Ad5 E1B but not with T24ras. AB - Expression of adenovirus type 5 E1A 12S is sufficient to immortalize primary baby rat kidney cells, but another viral or cellular oncogene, such as E1B or T24ras, is necessary for complete transformation. The regions of 12S sufficient for T24ras cotransformation have been well characterized and are located in the first exon. The second exon is dispensable for ras cotransformation, although it contains a region which appears to modulate the transforming phenotype. The same 12S first exon regions important in ras transformation are also necessary for E1B transformation. Analysis of an extensive series of second exon deletion and amino acid point mutations demonstrated that mutations affecting either the efficient nuclear localization and/or the immortalizing ability of the 12S protein also prevented cooperation with E1B. In general, the entire C-terminal half of 12S, including the nuclear localization signal, was necessary for efficient cotransformation with E1B. In addition to the differences between T24ras and E1B regarding 12S regions necessary for cotransformation, the characteristics of E1B cotransformed foci differed from those of T24ras. The E1B foci took longer to appear and had a much slower growth rate. No hypertransformed foci were produced with E1B cotransfections, and established E1A-E1B lines exhibited minimal growth in soft agar compared with that of E1A-T24ras lines. PMID- 7494323 TI - A novel non-mouse mammary tumor virus activation of the Int-3 gene in a spontaneous mouse mammary tumor. AB - In a mouse mammary tumor model system in which carcinogenic progression can be investigated, we have found a unique mutation of Int-3 associated with progression from premalignant lobular hyperplasia to tumor. Sequence analysis of the rearranged fragment revealed an insertion of an intracisternal type A particle (IAP) within the Int-3 gene. Int-3 is mutated frequently in mouse mammary tumor virus (MMTV)-induced mammary tumors by insertion of MMTV proviral DNA into this intragenic region. In these mutations, the insertion produces a chimeric Int-3 transcript encoding the cytoplasmic portion of the Int-3 protein driven by the MMTV long terminal repeat promoter. In this case, the IAP DNA was inserted in the opposite transcriptional orientation relative to Int-3; nevertheless, a similar chimeric RNA transcript driven by a cryptic promoter in the oppositely oriented 5' IAP long terminal repeat was generated. This is the first demonstration that an insertional mutation unrelated to MMTV activates an Int gene commonly associated with mammary tumorigenesis. PMID- 7494324 TI - Characterization of a neurologic disease induced by a polytropic murine retrovirus: evidence for differential targeting of ecotropic and polytropic viruses in the brain. AB - A variety of ecotropic murine leukemia viruses cause neurodegenerative disease. We describe here the clinical and histopathological features of a neurologic disease induced by a polytropic murine leukemia virus, FMCF98. Clinical disease was dominated by hyperexcitability and ataxia, and the histopathology was characterized primarily by astrocytosis and astrocytic degeneration. The viral envelope gene harbored the determinants of neurovirulence, since the chimeric virus Fr98E, which contained the envelope gene of FMCF98 on a background of the nonneurovirulent virus FB29, caused a similar disease. The disease caused by Fr98E differed from that induced by the coisogenic neurovirulent ecotropic virus FrCasE in clinical presentation, histopathology, and distribution of virus in the central nervous system. Since Fr98E contains a polytropic envelope gene and FrCasE contains an ecotropic envelope gene, these phenotypic differences appeared to be determined by envelope sequences and may reflect differences in virus receptor usage in the central nervous system. PMID- 7494325 TI - The leader peptide of Theiler's murine encephalomyelitis virus is a zinc-binding protein. AB - The leader (L) peptide is located in the amino-terminal part of the polyprotein of members of the Cardiovirus (which includes Theiler's murine encephalomyelitis virus) and Aphthovirus genera of picornaviruses. Although the function of L is unknown, strain DA of Theiler's murine encephalomyelitis virus with a mutation of L produces a cell-specific restricted infection. We now report that the DA L peptide is a metalloprotein and that zinc binds to a Cys-His motif that is conserved among cardioviruses. PMID- 7494326 TI - Lack of detection of negative-strand hepatitis C virus RNA in peripheral blood mononuclear cells and other extrahepatic tissues by the highly strand-specific rTth reverse transcriptase PCR. AB - To further explore the controversial potential for extrahepatic replication of hepatitis C virus (HCV), the highly strand-specific rTth method of reverse transcriptase PCR was used to examine sera, liver, peripheral blood mononuclear cells, and other extrahepatic tissues from HCV-infected chimpanzees and humans. Positive-strand HCV RNA was present in the liver at approximately 10-fold-higher levels than negative-strand HCV RNA. No negative-strand RNA was detected in peripheral blood mononuclear cells or other extrahepatic tissues despite the presence of abundant positive-strand RNA. These data demonstrate that within the limits of sensitivity of this highly strand-specific reverse transcriptase PCR method, no extrahepatic replication of HCV was detected. PMID- 7494327 TI - Susceptibility and signs associated with mouse adenovirus type 1 infection of adult outbred Swiss mice. AB - Adult Swiss outbred mice from two sources had a nearly 6,000-fold difference in susceptibility to mouse adenovirus type 1-induced disease. This difference was not attributable to differential organ tropism. Signs associated with mouse adenovirus type 1 infection that have not been previously reported are described at the clinical, gross pathological, and histological levels. PMID- 7494328 TI - Expression of ecotropic murine leukemia virus in the brains of C58/M, DBA2/J, and in utero-infected CE/J mice. AB - In C58 and AKR mice, endogenous N-tropic, ecotropic murine leukemia virus (MuLV) proviruses become activated in rare cells during embryogenesis. Resultant replication-competent progeny viruses then actively infect a large number of cells throughout the fetus, including cells in the developing central nervous system. By in situ hybridization analyses, we have assessed the presence of ecotropic MuLV RNA in the brains of C58 mice as a function of age. Only a few ecotropic MuLV-positive cells were observed in weanling mice, but the number of positive cells in the brain increased progressively with increasing age of the mice. Throughout the lives of the mice, the ecotropic MuLV RNA-positive cells were primarily located in well-defined white-matter tracts of the brain (commissura anterior, corpus callosum, fimbria hippocampi, optical tract, and striatum) and of the spinal cord. Cells of the subventricular zone also expressed ecotropic MuLV RNA, and in older mice a small number of positive cells were present in the grey matter. Infection of endogenous ecotropic MuLV provirus-less CE/J mice in utero with ecotropic MuLV clone AKR-623 resulted in the extensive infection of brain cells. The regional distribution of ecotropic MuLV RNA containing cells was the same as observed in the brains of C58 mice, in which cells became infected by endogenously activated virus, but the number of positive cells was higher. PMID- 7494329 TI - Transcriptional transactivation of parvovirus B19 promoters in nonpermissive human cells by adenovirus type 2. AB - The pathogenic human parvovirus B19 contains a promoter at map unit 6 (B19p6) of the viral genome, expression from which is largely restricted to human cells in the erythroid lineage, whereas a putative promoter at map unit 44 (B19p44) is inactive during a natural viral infection. Although nonerythroid human cells, such as HeLa and KB, allow expression from the B19p6 promoter but not from the B19p44 promoter following DNA-mediated transfection, little expression from the B19p6 promoter occurs following recombinant virus infection (S. Ponnazhagan, X. S. Wang, M.J. Woody, F. Luo, L.Y. Kang, M.L. Nallari, N.C. Munshi, S.Z. Zhou, and A. Srivastava, submitted for publication). However, significant expression from the B19p6 promoter as well as the B19p44 promoter could be detected in a human 293 cells line that expresses the adenovirus early gene products, suggesting that coinfection with adenovirus might mediate transcriptional transactivation of the B19 promoters in nonpermissive cells. Expression of the firefly luciferase reporter gene from the B19 promoters was evaluated either following plasmid transfection or following infection with the recombinant adeno-associated virus type 2 vectors. Both B19p6 and B19p44 promoters could be transactivated by coinfection with adenovirus in nonpermissive human cells, although the extent of transactivation of the B19p44 promoter was significantly lower than that of the B19p6 promoter. Expression of the adenovirus E1A proteins was necessary and sufficient for the observed transactivation of the B19 promoters. These studies further illustrate that the underlying molecular mechanisms of transactivation of parvovirus promoters in general by the adenovirus early proteins have similarities with those of the well-documented transactivation of the adeno associated virus type 2 promoters. PMID- 7494330 TI - Duck hepatitis B virus integrations in LMH chicken hepatoma cells: identification and characterization of new episomally derived integrations. AB - While the cytoplasmic phase of the hepadnavirus replication cycle is well understood, very little is known about the nuclear phase. In contrast to retroviruses, proviral integration is not required for hepadnavirus replication; however, some of the viral DNAs in the nucleus are diverted into an integration pathway. Under certain conditions these integrations function as carcinogenic agents. In order to study the integration process, we have utilized LMH-D2 cells, which replicate wild-type duck hepatitis B virus (DHBV), to develop the first protocol to detect and characterize integrations of DHBV originating from episomal viral DNAs. Contrary to expectations, our results showed that stable new integrations are readily detectable in subclones of LMH-D2 cells. Complete characterization of one integration revealed a single-genome-length integrant with the structure of double-stranded linear (DSL) DHBV DNAs which are produced by in situ priming during viral replication. The integration contained a terminal redundancy of 6 bp from the r region of the virus DNA minus strand as well as a direct repeat of 70 bp of cellular DNA. On the basis of the structure of the integrant and the cellular DNA target site, we propose a molecular model for the integration mechanism that has some similarities to that of retroviruses. Identification of DSL hepadnavirus DNA integration suggests the possibility that modified DSL viral DNAs may be the precursors to a class of simple, unrearranged hepadnavirus integrations. PMID- 7494331 TI - Immunization with recombinant varicella-zoster virus expressing herpes simplex virus type 2 glycoprotein D reduces the severity of genital herpes in guinea pigs. AB - Varicella-zoster virus (VZV) is an attractive candidate for a live-virus vector for the delivery of foreign antigens. The Oka vaccine strain of VZV is safe and effective in humans, and recombinant Oka VZV (ROka) can be generated by transfecting cells with a set of overlapping cosmid DNAs. By this method, the herpes simplex virus type 2 (HSV-2) glycoprotein D (gD2) gene was inserted into an intergenic site in the unique short region of the Oka VZV genome. Expression of gD2 in cells infected with the recombinant Oka strain VZV (ROka-gD2) was confirmed by antibody staining of fixed cells and by immunoblot analysis. Immune electron microscopy demonstrated the presence of gD2 in the envelope of ROka-gD2 virions. The ability of ROka-gD2 to protect guinea pigs against HSV-2 challenge was assessed by inoculating animals with three doses of uninfected human fibroblasts, fibroblasts infected with ROka VZV, or fibroblasts infected with ROka-gD2. Neutralizing antibodies specific for HSV-2 developed in animals immunized with ROka-gD2. Forty days after the third inoculation, animals were challenged intravaginally with HSV-2. Inoculation of guinea pigs with ROka-gD2 significantly reduced the severity of primary HSV-2 infection (P < 0.001). These experiments demonstrate that the Oka strain of VZV can be used as a live virus vector to protect animals from disease with a heterologous virus. PMID- 7494332 TI - Immortalization of human T cells expressing T-cell receptor gamma delta by herpesvirus saimiri. AB - Herpesvirus saimiri (HVS) has recently been shown to immortalize human CD4+ and CD8+ T cells expressing T-cell receptor alpha beta (TCR-alpha beta) with the maintenance of their original phenotypes and functional properties. However, the immortalization of human T cells expressing TCR-gamma delta by HVS has not been successful. Here we report that HVS can also infect and immortalize human T cells expressing TCR-gamma delta. Two human TCR-gamma delta+ T-cell clones, which continuously proliferated in interleukin-2-containing culture medium without any exogenous stimulation or addition of feeder cells for more than 8 months, were established by HVS infection. Morphologically, the HVS-transformed TCR-gamma delta+ T-cell clones were granular lymphocytes which exhibited wide-range HLA unrestricted cytotoxicity as untransformed TCR-gamma delta+ T cells. Their phenotypes and cytotoxic activities were not altered during long-term culture. The immortalization of human TCR-gamma delta+ T cells by HVS infection would be useful for functional analysis of this lymphocyte population, which is believed to play an important role in protection against various infectious diseases. PMID- 7494334 TI - Posttranslational signal peptidase cleavage at the flavivirus C-prM junction in vitro. AB - We have investigated the cleavages at the flavivirus capsid-prM protein junction in vitro. When expressed in the absence of the flavivirus proteinase, capsid and prM, which are separated by an internal signal sequence, exist as a membrane spanning precursor protein. Here we show the induction of posttranslational signal peptidase cleavage of prM by trypsin cleavage of a cytoplasmic region of this precursor protein. PMID- 7494333 TI - Diversity in virus populations from genital secretions and peripheral blood from women recently infected with human immunodeficiency virus type 1. AB - In order to develop a human immunodeficiency virus type 1 vaccine with global efficacy, it is important to evaluate the virus populations that are transmitted to individuals living in high-incidence areas. To determine the nature of the human immunodeficiency virus type 1 population transmitted to women during heterosexual contact, we examined the diversity of the proviral envelope gene in infected cells in both genital secretions and peripheral blood from six recently seroconverted Kenyan women. Heterogeneous virus populations were present in cervical secretions and/or peripheral blood shortly after seroconversion for five of six infected individuals, and tissue-specific variants were identified in several cases. PMID- 7494335 TI - CD8+ T-cell epitopes within the surface glycoprotein of a neurotropic coronavirus and correlation with pathogenicity. AB - CD8+ T cells with cytotoxic activity against the surface glycoprotein (S) of mouse hepatitis virus, strain JHM, have been identified in the central nervous system (CNS) of both acutely and chronically infected C57BL/6 mice. In this report, two specific epitopes recognized by these CNS-derived cells were identified, using a panel of peptides chosen because they conformed to the allele specific binding motif for MHC class I H-2Kb and H-2Db. The active peptides encompassed residues 510 to 518 (CSLWNGPHL, H-2Db) and 598 to 605 (RCQIFANI, H 2Kb). Both epitopes are located within the region of the S protein previously shown to be prone to deletion after passage in animals. These deleted strains are generally less neurovirulent than the wild-type virus but still are able to cause demyelination. Since C57BL/6 mice become persistently infected more commonly than many other strains of mice, these data are consistent with a role for CD8+ T-cell escape mutants in the pathogenesis of the demyelinating disease. This is the first report of CD8+ T-cell epitope localization within the S protein, the protein most strongly implicated thus far in pathogenesis in the host. PMID- 7494336 TI - Complete genetic characterization and analysis of isolation of Sin Nombre virus. AB - This study reports completion of the genetic characterization of the entire genome of Sin Nombre (SN) virus (NMH10) detected in autopsy tissues from a patient who died of hantavirus pulmonary syndrome (HPS). The large (L) genome segment was found to be 6,562 nucleotides in length and encoded a putative L polymerase that was 2,153 amino acids in length. No evidence of segment reassortment with other well-characterized hantaviruses was obtained. The sequence of the entire S, M, and L genome segments of SN virus (strain NMR11) isolated from a mouse (trapped in the residence of the patient infected with SN virus [NMH10]) by passage two times in Peromyscus maniculatus and then by five passages in E6 Vero cells was determined and compared with that of the virus detected in autopsy tissues. Only 16 nucleotide differences were detected between the virus genomes, and none of these resulted in virus protein amino acid substitutions. Determination of the exact 5'- and 3'-terminal sequences of all genome segments of SN virus and representatives of other serologic groups in the Hantavirus genus, family Bunyaviridae, showed the existence of conserved nucleotide domains that may be involved in important regulatory mechanisms, such as RNA encapsidation, polymerase binding, and control of transcription and replication. PMID- 7494337 TI - Molecular linkage of hantavirus pulmonary syndrome to the white-footed mouse, Peromyscus leucopus: genetic characterization of the M genome of New York virus. AB - The complete M segment sequences of hantaviruses amplified from tissues of a patient with hantavirus pulmonary syndrome in the northeastern United States and from white-footed mice, Peromyscus leucopus, from New York were 99% identical and differed from those of Four Corners virus by 23%. The serum of this patient failed to recognize a conserved, immunodominant epitope of the Four Corners virus G1 glycoprotein. Collectively, these findings indicate that P. leucopus harbors a genetically and antigenically distinct hantavirus that causes hantavirus pulmonary syndrome. PMID- 7494338 TI - Biological characterization of nef in long-term survivors of human immunodeficiency virus type 1 infection. AB - We have previously shown that there were no gross deletions or obvious sequence abnormalities within nef of human immunodeficiency virus type 1 (HIV-1) in the 10 long-term survivors studied (Y. Huang, L. Zhang, and D. D. Ho, J. Virol. 69:93 100, 1995). Here we extend our study to examine these nef alleles in a functional context. Using a new technique, termed site-directed gene replacement, we have precisely replaced the nef of an infectious molecular clone, HIV-1HXB2, with nef alleles derived from 10 long-term survivors as well as from a patient with AIDS. The replication properties of these chimeric viruses demonstrated that the nef alleles derived from long-term survivors neither significantly increased nor decreased viral replication, compared with the nef allele of Nef+ HIV-1HXB2 and that derived from a patient with AIDS. However, Nef+ viruses always replicated faster than virus lacking nef. Moreover, single-cell infection analysis by the MAGI assay showed that these chimeric viruses, as well as Nef+ HIV-1HXB2, were more infectious than Nef- HIV-1HXB2 was. Therefore, we conclude that the genotypic and phenotypic features of nef are not likely to account for the nonprogression of HIV-1 infection in the 10 cases studied, unless the function of the nef gene in vivo is not accurately reflected by the in vitro assays we used. PMID- 7494339 TI - The in vivo antiviral activity of interleukin-12 is mediated by gamma interferon. AB - The injection of 20 ng of mouse interleukin-12 (IL-12) protects mice from a lethal infection with encephalomyocarditis virus. In vitro, an anti-gamma interferon (anti-IFN-gamma) monoclonal antibody but not an anti-IL-12 monoclonal antibody neutralizes the antiviral activity present in the supernatants of splenocytes stimulated with IL-12. Finally, IL-12 fails to protect 129 Sv/Ev IFN gamma R0/0 mice against encephalomyocarditis virus infection. These results demonstrate that IL-12 exerts its antiviral activity through the induction of endogenous IFN-gamma. PMID- 7494340 TI - Configuration of latent varicella-zoster virus DNA. AB - The configuration of latent varicella-zoster virus (VZV) DNA was analyzed by PCR. Template DNA for both internal and terminal VZV primers was present in a 1:1 ratio in ganglionic DNA, compared with a 15:1 ratio in DNA extracted from VZV virions, indicating that the VZV genomic termini are adjacent in latently infected human ganglia. PMID- 7494341 TI - Epstein-Barr virus induction of recombinase-activating genes RAG1 and RAG2. AB - In experimental B-cell infections, Epstein-Barr virus induced sustained expression of V(D)J recombinase-activating genes RAG1 and RAG2, whose aberrant activity has been implicated in chromosomal translocations in B-cell neoplasms. In cell lines in which RAG1 and RAG2 were detected, virus integrated into cellular DNA rather than assumed the configuration of extrachromosomal episomes. Expression of the Epstein-Barr virus nuclear antigen 1 in transient transfection assays was sufficient to induce both recombinase-activating genes. PMID- 7494342 TI - Molecular and functional characterization of turkey interferon. AB - The turkey interferon (TkIFN) gene encodes a signal peptide and a mature protein of 30 and 162 amino acids, respectively. TkIFN mRNA expression was induced by reoviral double-stranded RNA in fibroblasts. The recombinant TkIFN protein possessed species-specific antiviral activity and in synergy with lipopolysaccharide (LPS) induced bone marrow macrophages to produce nitric oxide (NO). LPS or TkIFN alone did not induce bone marrow macrophages to produce significant amounts of NO, which showed that TkIFN provided one of the two signals necessary to induce NO production in turkey macrophages. Unlike the anti inflammatory nature of mammalian alpha/beta IFNs, TkIFN augmented the LPS-induced expression of interleukin-8, a proinflammatory cytokine. This finding suggests a role for TkIFN in inflammatory conditions. PMID- 7494343 TI - Role of CD4 endocytosis in human immunodeficiency virus infection. AB - We have analyzed the role of CD4 endocytosis in human immunodeficiency virus (HIV) entry by measuring the infection of HeLa cells expressing various CD4 constructs with endocytosis rates of between 0.2 and 30%/min in a quantitative infectious focus assay. For a number of laboratory-adapted HIV-1 and HIV-2 strains, the highest levels of infection were found on cells with very limited CD4 endocytosis, while cells with efficient CD4 uptake were only poorly infectable, suggesting that CD4 internalization is not required for HIV entry. This was confirmed in a modified assay involving prebinding of HIV-1LAI to HeLa CD4 cells at 4 degrees C, synchronized virus entry during warming to 37 degrees C, and neutralization of virions remaining at the cell surface with anti-V3 loop antibodies. Warming cells in hypertonic medium inhibited CD4 endocytosis but did not affect the rate or the extent of infection. These studies confirm that HIV infection does not require endocytosis and that laboratory-adapted virus strains can enter HeLa-CD4 cells by fusion at the plasma membrane. PMID- 7494344 TI - The Epstein-Barr virus nuclear antigen leader protein associates with hsp72/hsc73. AB - Epstein-Barr virus nuclear antigen leader protein (EBNA-LP) is important for primary B-lymphocyte growth transformation. We now demonstrate that the W repeat encoded domain of EBNA-LP significantly associates with proteins of the heat shock protein 70 family (hsp72/hsc73). hsp72/hsc73 may mediate the previously observed interaction between EBNA-LP and the retinoblastoma protein or p53. PMID- 7494346 TI - Roles of different T-cell subsets in control of herpes simplex virus infection determined by using T-cell-deficient mouse-models. AB - Herpes simplex virus infection of the scarified dermis results in infection of the nervous system and, subsequently, a cutaneous lesion in the innervated dermatome. We compared the pathogenesis of such zosteriform lesions in mice lacking or severely depleted of CD4+ or CD8+ T cells because of targeted gene disruption. Mice without CD4+ cells showed markedly increased susceptibility, whereas beta 2 microglobulin knockout mice lacking CD8+ T cells were as resistant to challenge as were immunocompetent mice with the same genetic background. Our results demonstrate that CD4+ T cells are of primary importance in the control of herpes simplex virus infections of the skin and nervous system. PMID- 7494345 TI - An aspartic acid at amino acid 108 is required to rescue infectious virus after transfection of a poliovirus cDNA containing a CGDD but not SGDD amino acid motif in 3Dpol. AB - The poliovirus RNA-dependent RNA polymerase (3Dpol) contains a region of homology centered around the amino acid motif YGDD (amino acids 326 to 329), which has been postulated to be involved in the catalytic activity of the enzyme. Previous studies from this laboratory have used oligonucleotide site-directed mutagenesis to substitute the tyrosine amino acid at this motif with other amino acids (S. A. Jablonski and C. D. Morrow, J. Virol. 67:373-381, 1993). The viruses recovered with 3Dpol genes with a methionine mutation also contained a second mutation at amino acid 108 resulting in a glutamic acid-to-aspartic acid change (3D-E-108 to 3D-D-108) in the poliovirus RNA polymerase. On the basis of these results, we suggested that the amino acid at position 108 might interact with the YGDD region of the poliovirus polymerase. To further investigate this possibility, we have constructed a series of constructs in which the poliovirus RNA polymerases contained a mutation at amino acid 108 (3D-E-108 to 3D-D-108) as well as a mutation in which the tyrosine amino acid (3D-Y-326) was substituted with cysteine (3D-C-326) or serine (3D-S-326). The mutant 3Dpol polymerases were expressed in Escherichia coli, and in vitro enzyme activity was analyzed. Enzymes containing the 3D-D-108 mutation with the wild-type amino acid (3D-Y-326) demonstrated in vitro enzyme activity similar to that of the wild-type enzyme containing 3D-E-108. In contrast, enzymes with the 3D-C-326 or 3D-S-326 mutation had less in vitro activity than the wild type. The inclusion of the second mutation at amino acid 3D-D-108 did not significantly affect the in vitro activity of the polymerases containing 3D-C-326 or 3D-S-326 mutation. Transfections of poliovirus cDNAs containing the substitution at amino acid 326 with or without the second mutation at amino acid 108 were performed. Consistent with previous findings, we found that transfection of poliovirus cDNAs containing the 3D-C-326 or 3D-S-326 mutation in 3Dpol did not result in the production of virus. Surprisingly, transfection of the poliovirus cDNAs containing the 3D-D 108/C-326 double mutation, but not the 3D-D-108/S-326 mutation, resulted in the production of virus. The virus obtained from transfection of polio-virus cDNAs containing 3D-D-108/C-326 mutation replicated with kinetics similar to that of the wild-type virus. RNA sequence analysis of the region of the 3Dpol containing the 3D-C-326 mutation revealed that the codon for cysteine (UGC) reverted to the codon for tyrosine (UAC). The results of these studies establish that under the appropriate conditions, poliovirus has the capacity to revert mutations within the YGDD amino acid motif of the poliovirus 3Dpol gene and further strengthen the idea that interaction between amino acid 108 and the YGDD region of 3Dpol is required for viral replication. PMID- 7494347 TI - Association of reovirus outer capsid proteins sigma 3 and mu 1 causes a conformational change that renders sigma 3 protease sensitive. AB - Association of the reovirus proteins sigma 3 and mu 1 influences viral entry, initiation of outer capsid assembly, and modulation of the effect of sigma 3 on cellular translation. In this study, we have addressed whether structural changes occur in sigma 3 as a result of its interaction with mu 1. Using differences in protease sensitivity to detect conformationally distinct forms of sigma 3, we showed that association of sigma 3 with mu 1 caused a conformational change in sigma 3 that converted it from a protease-resistant to a protease-sensitive structure and occurred posttranslationally. The effect of mu 1 on the structure of sigma 3 was stoichiometric. Our results are consistent with a model in which sigma 3's association with mu 1 shifts its function from translational control to assembly of an outer capsid in which sigma 3 is folded into the protease sensitive conformation that is required for its cleavage during the next round of infection. PMID- 7494348 TI - Clinical research: primary objective of the American Venous Forum. PMID- 7494349 TI - Limited range of motion is a significant factor in venous ulceration. AB - PURPOSE: Calf muscle pump dysfunction is a recognized factor in chronic venous insufficiency (CVI). We investigated the hypothesis that limbs with CVI have a reduced ankle range of motion (ROM) that may be responsible for the poor calf pump function associated with venous ulceration. METHODS: Ankle ROM and calf pump function were assessed in 32 limbs of 26 adult men. Limbs were selected on the basis of clinical presentation: normal (n = 6 limbs), class 1 or 2 CVI with no history of ulceration (n = 9 limbs), class 3 CVI with healed ulceration (n = 9 limbs), and class 3 CVI with active ulceration (n = 8 limbs). ROM was determined by goniometry during maximal plantar flexion and dorsiflexion of the ankle. Calf pump function was determined by air plethysmographic measurement of ejection fraction (EF) and residual volume fraction (RVF). RESULTS: Ankle ROM was significantly (p < 0.05) reduced in each CVI group compared with age-matched control subjects, because of decreases in both plantar flexion and dorsiflexion. Calf pump function was significantly impaired (decreased EF and increased RVF) in ulcerated limbs. ROM was significantly correlated to EF and RVF. Impairment of ROM and calf pump function was associated with deterioration in the clinical classification of venous disease. CONCLUSIONS: Limbs with CVI have a limited ankle ROM that decreases with increasing severity of clinical symptoms. This decreased ROM is associated with, and may contribute to, poor calf pump function. PMID- 7494350 TI - Venous valvular reflux in veins not involved at the time of acute deep vein thrombosis. AB - PURPOSE: The aim of this study was to determine whether, in lower extremities with documented episodes of acute deep venous thrombosis (DVT), incompetence develops in veins that were not the site of thrombosis. METHODS: Patients were monitored with serial duplex ultrasonography at 1 day, 1 week, 1, 3, 6, 9, and 12 months, and then annually after detection of acute DVT. The following venous segments were analyzed: common femoral, greater saphenous, proximal superficial femoral, deep femoral, popliteal, and posterior tibial. The incidence of reflux development in both thrombosed and uninvolved segments was determined. Reflux was categorized as either transient or permanent. RESULTS: A total of 227 limbs in 188 patients were serially studied. Mean follow-up was 19.9 months (range 1 to 88 months). Overall, 403 of the 1423 segments (28.3% +/- 2.3%) developed reflux during the study, of which 118 (29.3% +/- 4.4%) had no prior or concurrent history of thrombosis. Considering only the segments that developed incompetence, the percent without prior thrombosis at each level was as follows: common femoral vein (40.0%), greater saphenous vein (53.1%), deep femoral vein (20.6%), proximal superficial femoral vein (23.9%), popliteal vein (8.9%), and posterior tibial vein (31.9%). Valvular insufficiency developing in segments uninvolved with thrombus was more likely to be transient (40.2%) than was the reflux in thrombosed segments (22.6%). This difference was statistically significant (p < 0.05). CONCLUSIONS: Permanent venous valvular damage can occur in the absence of thrombosis after DVT. Reflux in uninvolved venous segments has a different anatomic distribution and is more likely to be transient than the incompetence associated with thrombosis. PMID- 7494351 TI - A noninvasive screening test to detect "critical" deep venous reflux. AB - PURPOSE: The venous filling index (VFI) was introduced as a noninvasive measure of venous valvular reflux. Because it is quantitative, identifying increasingly severe reflux should be possible. The purpose of this study is to evaluate the sensitivity and predictive value of the VFI as a predictor of phlebographically demonstrated "critical" venous reflux. METHODS: Thirty-one limbs with suspected venous insufficiency underwent both descending phlebography and air plethysmography. Nine limbs had deep venous reflux ending above the knee on descending phlebography (group 1), and 22 limbs had "critical" deep reflux to the below-knee level (group 2). The VFI, ejection fraction, and residual volume fraction were calculated, and a tourniquet was used to distinguish superficial from deep venous reflux. RESULTS: Three of six group 2 limbs with a VFI less than 7 had obliteration of their iliac veins. Tourniquet application improved the VFI in 13 limbs, all of which showed either phlebographic or venous duplex evidence of greater saphenous incompetence. CONCLUSIONS: A VFI greater than 7 showed a 73% sensitivity and 100% positive predictive value of identifying "critical venous reflux." The VFI may underestimate the degree of reflux in patients with proximal venous obstruction. Improvement of venous hemodynamics with tourniquet application suggests that venous reflux might be improved in some patients by correcting superficial venous insufficiency. PMID- 7494352 TI - Gender bias in use of venous ultrasonography for diagnosis of deep venous thrombosis. AB - PURPOSE: We observed that ultrasound examinations for deep venous thrombosis (DVT) were more frequently requested for women than for men in our vascular laboratory serving a general outpatient population and referral 774-bed hospital. Because existing literature presents conflicting information about sex differences in occurrence of DVT, we investigated correlation in our population with positive ultrasound study results and risk factors for DVT. METHODS: In 13 months, 2055 ultrasound examinations for DVT were requested. Of these, 300 patients (15%) were categorized in four subgroups: 75 ultrasonography-negative men, 75 ultrasonography-negative women, 75 ultrasonography (DVT)-positive men, and 75 ultrasonography (DVT)-positive women for risk factor analysis. RESULTS: Women comprised 64% (1311 of 2055) and men 36% (744 of 2055) of ultrasound examinations requested, but men had significantly higher incidence of DVT positive ultrasonography results (101 of 744 [14%]) compared with women (118 of 1311 [9%]) (p = 0.002 by chi-square testing). There were no significant sex differences in conventional DVT risk factors and no difference in aggregate number of risk factors. The anatomic distribution of DVT was the same in men as in women. Among those having negative ultrasonography results, significantly more outpatient examinations were performed in women (p = 0.018 by t testing). CONCLUSIONS: Gender bias exists in use of ultrasonography for diagnosis of DVT. The greater incidence of women undergoing venous ultrasonography is not explained by higher prevalence of DVT risk factors or of higher occurrence of positive ultrasound examination results. Further investigation is needed to determine whether these differences indicate underuse of ultrasonography in men or overuse in women. PMID- 7494353 TI - Contralateral duplex scanning for deep venous thrombosis is unnecessary in patients with symptoms. AB - PURPOSE: Bilateral lower extremity venous duplex scanning for acute deep venous thrombosis (DVT) has been advocated because of the high incidence of occult contralateral leg involvement. We investigated the clinical necessity of such a policy. METHODS: The results from 2996 venous duplex studies performed during the past 2 years were retrospectively reviewed. A total of 1694 of these scans were performed on patients with symptoms, of whom 248 (15%) were found to have an acute DVT. Symptoms were limited to one side in 198 patients, whereas bilateral complaints were noted in 50 patients. RESULTS: Among the patients with symptoms of acute DVT, 72 (29%) had bilateral involvement. Bilaterality was more likely in patients with bilateral symptoms than in those with only unilateral symptoms (56% vs 22%; p < 0.005). Of the patients with unilateral symptoms and bilateral DVT, all of them had either acute (80%) or acute and chronic (20%) thrombosis in the symptomatic leg. The contralateral asymptomatic limb had fewer acute and more chronic DVT (41% and 55%, respectively). No patient from the entire group admitted with symptoms had an acute DVT in the asymptomatic limb without a concomitant acute DVT in the symptomatic leg. Unilateral scanning would decrease the examination time by 21% and potentially increase total reimbursement for symptomatic venous scans by 9% compared with routine bilateral duplex scanning. CONCLUSIONS: Although bilateral involvement is frequent in patients with symptoms of acute DVT, treatment in these patients is not altered by this finding. We conclude that contralateral venous scanning in patients with unilateral symptoms is not clinically indicated and that unilateral scanning would result in improved cost-efficiency for vascular laboratories. PMID- 7494354 TI - Vascular laboratory personnel on-call: effect on patient management. AB - PURPOSE: A prospective study was undertaken of all vascular laboratory tests performed by the "on-call" technologist during a 3-year time period. METHODS: Technologists take call on a rotating basis. All patients evaluated had symptoms. History and risk factor data were collected at the time of patient encounter. RESULTS: Studies were initiated from the emergency department 56% of the time, from inpatient examinations 33% of the time, and from outpatient examinations 11% of the time. Ninety percent (n = 440) of the studies obtained were venous duplex scans (VDS). Fifty-eight percent (257 of 440) of the studies were performed on weekends. Thirty-two percent of the studies resulted in admission of the patient. Of the 440 VDS obtained, 51% (224 of 440) identified some type of disease. Acute deep venous thrombosis (DVT) was diagnosed in 15% (67 of 440), acute superficial venous thrombosis (SVT) in 4% (17 of 440), acute DVT and SVT in 7% (31 of 440), chronic DVT in 4% (17 of 440), chronic SVT in 4% (16 of 440), and chronic DVT and SVT in 1% (3 of 440); 2% (9 of 440) of the DVTs were of indeterminate age. Other diseases were identified in 22% (95 of 440), including popliteal cysts, inguinal lymphadenopathy, and soft tissue edema. CONCLUSIONS: Having an "on-call" vascular technologist has allowed patients with acute venous thrombosis to be triaged and admitted, if necessary, in a timely fashion. Those patients who have symptoms and a negative VDS result are able to avoid unnecessary hospitalization and treatment. The expense of this service is easily justified by the savings of unnecessary hospital days. PMID- 7494355 TI - Limited B-mode venous imaging versus complete color-flow duplex venous scanning for detection of proximal deep venous thrombosis. AB - PURPOSE: The purpose of this study was to compare the accuracy of a limited B mode compression technique (BMCT) with a complete color-flow duplex venous examination (CDVE) for the detection of proximal deep vein thrombosis (DVT). METHODS: We prospectively studied 72 patients (20 men and 52 women) for DVT. Two technologists blinded to each other performed either BMCT or CDVE independently. The BMCT is an abbreviated technique compressing two sites per limb. One site was the saphenofemoral junction including the superficial femoral and deep femoral vein confluence; the other was the saphenopopliteal junction including tibial vein confluence. Total limbs studied were 144. CDVE was considered the gold standard for purposes of sensitivity, specificity, and accuracy. RESULTS: The technical failure rate of BMCT was three of 144. In all technically satisfactory examinations, the BMCT result was positive in 15 of 14 limbs, and the CDVE result was positive in 13. Sensitivity of BMCT was 100%, specificity was 98%, and overall accuracy was 99%. There were two false-positive results with BMCT; both were cases of popliteal veins deep to the artery leading to difficulty in compression. The BMCT was able to detect chronic thrombus, floating thrombus, and small thrombus behind femoral vein valve cusps. CONCLUSION: These data suggest that BMCT is a rapid, acceptable, alternative technique for detecting proximal DVT. In cases of equivocal or positive findings, the spectral and color-flow Doppler examination should be used to confirm the results. PMID- 7494356 TI - Propagation, rethrombosis and new thrombus formation after acute deep venous thrombosis. AB - PURPOSE: The purpose of this study was to determine the incidence, timing, and outcome of further thrombotic events after an initial episode of acute deep venous thrombosis. METHODS: Venous thrombi in 204 lower extremities (177 patients) were monitored with duplex ultrasonography at intervals of 1 day, 7 days, 1 month, every 3 months for 1 year, and yearly thereafter. RESULTS: Among initially involved extremities, propagation to new segments occurred in 61 (30%) and rethrombosis occurred in 63 (31%). Both propagation and rethrombosis, in different segments, occurred in 27 (13%) extremities. New thrombi were also noted in nine (6%) initially uninvolved extremities. These events were not associated with identifiable clinical risk factors, although extremities with rethrombosis were more extensively involved at presentation. Propagation in initially involved extremities was an early event, occurring within a median of 40 days in all segments. New thrombotic events in initially uninvolved extremities and rethrombosis occurred as later events. The development of reflux was significantly more common among all initially uninvolved segments to which thrombus extended and among mid and distal superficial femoral and popliteal artery segments with rethrombosis. CONCLUSIONS: Recurrent thrombotic events are common after acute deep venous thrombosis and adversely affect the ultimate development of valvular incompetence. Their occurrence is unrelated to recognized clinical risk factors and can occur despite standard anticoagulation measures. PMID- 7494357 TI - Modifications of plasma levels of tissue factor pathway inhibitor and endothelin 1 induced by a reverse Trendelenburg position: influence of elastic compression- preliminary results. AB - PURPOSE: The purpose of this study was to assess the effects of the passive 45 degree reverse Trendelenburg position and graduated compression stockings (GCS) on plasma tissue factor pathway inhibitor (TFPI) and endothelin-1 levels in a group of volunteers. METHODS: Ten healthy subjects lay on an examining table for 30 minutes while baseline measurements were made. The table was then tilted to a 45-degree upright position for 60 minutes, and measurements were repeated. On a different day, subjects were tilted again, but on this occasion they wore thigh length GCS. Blood was drawn before and 60 minutes after tilting during both sessions, and plasma TFPI and endothelin-1 were obtained. Cross-sectional areas of the calf medial gastrocnemius vein, before and after tilting, were measured by a duplex scanner. RESULTS: Upright tilting induced a significant dilation of the medial gastrocnemius veins that was partially corrected by the use of elastic stockings. Similarly, endothelin-1 levels significantly increased after tilting. The use of GCS did not modify these differences. On the other hand, although TFPI levels were not affected by tilting without stockings, they were significantly elevated after tilting when GCS were used. CONCLUSIONS: Upright passive tilting induces significant dilation of the deep calf veins and is associated with a significant increase in plasma levels of endothelin-1, whereas TFPI levels remain unchanged. The use of elastic stockings reduces the degree of calf distention but does not prevent an increase in endothelin-1. TFPI levels are significantly increased after tilting when GCS are used. This might represent a previously unknown mechanism of action of elastic stockings with interesting potential for deep vein thrombosis prophylaxis. More studies are warranted in a larger series to confirm these results. PMID- 7494358 TI - The tissue plasminogen activator and urokinase response in vivo during natural resolution of venous thrombus. AB - PURPOSE: The aim of this study was to measure the distribution of endogenous plasminogen activators during thrombolysis with an endothelial-conserving model of laminated thrombosis. METHODS: Thrombi were raised in the inferior vena cava of rats with thrombin and flow reduction. The thrombi, adjacent vein wall, and distant veins (the superior vena cava) were removed at intervals from 1 hour to 21 days from formation and then cryohomogenized and assayed with specific bioimmunoassays for tissue-type (t-PA) and urokinase-type plasminogen activators (u-PA). RESULTS: The measured t-PA activity of the vein wall around the thrombus was reduced compared with the control inferior vena cava at 4 days. Both the u-PA and t-PA content of the thrombus increased progressively during thrombolysis. The t-PA activity increased significantly in the distant vein walls in the animals with thrombi. Immunocytochemistry and in situ hybridization localized the t-PA to a mononuclear cell infiltrate and showed up-regulation of mRNA for rat t-PA in these monocytes. CONCLUSIONS: The local plasminogen activator response was predominantly within the thrombus itself. Increased t-PA activity was additionally found in distant veins but was reduced in the vessel wall adjacent to the thrombus. This is the first report to show that u-PA activity is increased within organizing thrombus in vivo and that most of the t-PA activity is localized to a monocyte infiltrate. PMID- 7494359 TI - Diminished mononuclear cell function is associated with chronic venous insufficiency. AB - PURPOSE: With clinical progression of chronic venous insufficiency (CVI), dermal infiltration of mononuclear cells increases. Because these cells regulate chronic inflammatory responses and modulate wound healing, cellular dysfunction could explain alterations in wound healing with CVI. The purpose of this study was to determine whether monocytes in patients with CVI are dysfunctional. METHODS: Mononuclear cell function was measured as the degree of proliferation in response to a mitogenic challenge. Fifty patients were separated into four groups: group 1, 14 patients with normal limbs; group 2, 10 patients with class 2 CVI; group 3, 15 patients with active venous ulcers; group 4, 11 patients with healed venous ulcers and current evidence of lipodermatosclerosis. Duplex scanning and air plethysmography correlated with the clinical classification of CVI. Systemically circulating monocytes and lymphocytes were obtained by antecubital venipuncture from groups 1 to 4. Cells were cultured in the presence of staphylococcal enterotoxins A, B, C1, D, and E (mitogens) at 1, 8, 31, and 125 micrograms/well on the basis of previous dose-response experiments. Phytohemagglutinin (PHA), 5 micrograms/well, served as a control mitogen. The dose-response curves indicated that 8 micrograms/well elicited the greatest degree of cell proliferation. Proliferative responses at 8 micrograms/well were analyzed for statistical significance among groups 1 to 4. Comparisons among groups were performed by use of the nonparametric Mann Whitney U post tests and a one-tailed unpaired t test. Results were considered significant at p < or = 0.05. RESULTS: Proliferative responses to PHA indicate that lymphocytes and monocytes from patients with CVI are not globally depressed. However, patients in group 2 did not exhibit the same degree of proliferation to PHA as did groups 1, 3 and 4. Proliferative responses between groups 2 and 1 (44.38 +/- 43.9 vs 118.87 +/- 27.1, p < or = 0.05) and groups 2 and 3 (44.38 +/- 43.9 vs 105.95 +/- 60.99, p < or = 0.05) were significant. Challenges with staphylococcal enterotoxin A and B reveal significant diminution of proliferative responses in groups 2 (42.73 +/- 11.55, p < or = 0.05) and 3 (45.57 +/- 9.1, p < or = 0.05) and groups 3 (36.81 +/- 6.9, p < or = 0.05) and 4 (35.04 +/- 7.5, p < or = 0.05), compared with staphylococcal enterotoxin A controls (68.68 +/- 9.9) and staphylococcal enterotoxin B controls (66.25 +/- 13.56), respectively. A trend of diminished mononuclear cell function with progression of CVI was observed with staphylococcal enterotoxins B, C1, D, and E, strongly suggesting biologic significance. Furthermore, patients with lipodermatosclerosis uniformly exhibited the poorest proliferative responses. CONCLUSIONS: Deterioration of mononuclear cell function is associated with CVI. A trend of diminishing proliferative responses with clinical disease progression is observed and suggests biologic significance. The decreased capacity for mononuclear cell proliferation in response to various challenges may manifest itself clinically as poor and prolonged wound healing. PMID- 7494360 TI - Flush saphenofemoral ligation and multiple stab phlebectomy preserve a useful greater saphenous vein four years after surgery. AB - PURPOSE: Flush saphenofemoral ligation, with multiple cosmetic stab phlebectomy, successfully treats saphenofemoral reflux and superficial varicosities. The long term effect on the residual greater saphenous vein (GSV) requires evaluation. METHODS: Noninvasive triplex ultrasound assessment of the residual GSV for potential use as a vein graft was used. We evaluated the independent objective and subjective cosmetic outcomes. We also used analysis to determine the correlation between GSV reflux, symptoms, and cosmetic outcome. RESULTS: Seventy two limbs were assessed at a median follow-up of 4 years (interquartile range 3 to 5 years). Fifty-nine limbs had patent GSVs above and below the knee. The mean length patent was 51 cm, and the mean internal diameters above and below the knee were 3.6 and 2.8 mm, respectively. In 65% of cases the vein had suitable ultrasonic characteristics, suggesting that it would be suitable for use as a conduit for a vascular bypass graft. Sixty-two limbs were considered a symptomatic success and 61 an objective cosmetic success. There was no correlation between the symptomatic and cosmetic outcome and the presence of superficial GSV reflux (linear regression analysis). CONCLUSIONS: These findings support the effectiveness of the procedure of flush saphenofemoral ligation and multiple stab phlebectomy with regard to cosmetic and symptomatic outcome and may preserve the GSV for use as an autologous bypass graft. PMID- 7494361 TI - Comparison of streptokinase, urokinase, and recombinant tissue plasminogen activator in an in vitro model of venous thrombolysis. AB - PURPOSE: Presumed differences in the thrombolytic activity and fibrinolytic specificity of the three commonly used thrombolytic agents, streptokinase, urokinase, and recombinant tissue plasminogen activator (rt-PA), are based on clinical study results, where variability renders meaningful comparisons difficult. An in vitro model of catheter-directed venous thrombolysis was used to compare the three agents. METHODS: Retracted iodine 125-radiolabeled clots that simulate those observed in the venous system were infused with thrombolytic agents at doses analogous to those used clinically. Perfusion with heparinized, whole human blood was undertaken for 60 minutes, measuring the efficacy of thrombolysis through serial quantification of radio tracer released into the circuit. Fibrinolytic specificity was determined by following decrements in perfusate fibrinogen concentration. RESULTS: Streptokinase was the agent associated with the slowest rate of clot lysis (p = 0.01 vs urokinase and rt-PA). Urokinase was associated with an intermediate rate of lysis but appeared to be the agent with the greatest degree of fibrinolytic specificity (p = 0.02 vs streptokinase, p = 0.05 vs rt-PA). Although rt-PA was associated with improved efficacy early in the perfusions, the differences between rt-PA and urokinase dissipated after 30 minutes. CONCLUSIONS: These laboratory observations suggest that urokinase may be the most appropriate agent for catheter-directed venous thrombolysis, offering an advantageous compromise between fibrinolytic specificity and thrombolytic speed. PMID- 7494362 TI - Venous hemodynamics during impulse foot pumping. AB - PURPOSE: This study was designed to measure the effect of intermittent pneumatic compression of the plantar venous plexus on popliteal vein (PV) and common femoral vein (CFV) velocities measured by duplex ultrasound scanning. METHODS: Thirty lower limbs in 15 healthy volunteers had venous duplex scanning measurement of PV and CFV velocities before and during foot pumping with an arteriovenous impulse foot pump system. Venous velocities were measured at two pump pressure settings (100 mm Hg, 200 mm Hg) and during two pump impulse durations (short = 1 second, normal = 3 seconds). All limbs were examined with the subjects in the supine position, and then measurements were repeated with subjects in the 15-degree reverse Trendelenburg position. The mean maximum venous velocity (MVV) produced by foot pumping was compared with resting venous velocity at each anatomic location and for each technologic variable. RESULTS: Impulse foot pumping produced a statistically significant increase in MVV in both the PV and the CFV compared with resting velocities. This significant increase was observed for both pressure settings and both impulse durations, and no differences produced by these two individual variables could be detected. The increase in MVV produced by foot pumping was similar for limbs in the supine position and those examined in the reverse Trendelenburg position. The percentage increase in MVV produced by foot pumping was significantly higher in the PV than in the CFV. CONCLUSIONS: Intermittent pneumatic compression of the plantar venous plexus produces measurable increases in venous outflow from the lower limbs of normal subjects. This study seems to justify further evaluation of the effectiveness of this technique for mechanical deep venous thrombosis prophylaxis in selected high-risk patient groups. PMID- 7494363 TI - Phlegmasia complicating prophylactic percutaneous inferior vena caval interruption: a word of caution. AB - PURPOSE: The purpose of this study was to evaluate the incidence of thrombotic complications in patients with deep vein thrombosis (DVT) who were treated with percutaneous inferior vena caval interruption in place of anticoagulation. METHODS: A retrospective review of all percutaneously placed inferior vena cava filters for 1 year, August 1993 through July 1994, was performed. RESULTS: Thirty three percutaneous inferior vena cava filters were placed in 32 patients. The underlying disease was pulmonary embolism in 15 (47%) and DVT in 17 (53%) patients. Of patients with pulmonary embolism, 11 had a documented DVT, and four were not evaluated for DVT. There were 14 men and 18 women, with a mean age of 63.5 years (range 24 to 93 years). Indications for vena caval interruption were recurrent pulmonary embolism with therapeutic anticoagulation (n = 2 [6%]), prophylactic insertion with documented pulmonary embolism and therapeutic anticoagulation (n = 8 [25%]), documented pulmonary embolism and absolute contraindication to anticoagulation (n = 5 [16%]), documented DVT and absolute contraindication to anticoagulation (n = 2 [6%]), prophylactic insertion with documented DVT and therapeutic anticoagulation (n = 5 [16%]), and documented DVT with relative contraindication to anticoagulation (n = 10 [31%]). Of the 32 patients with inferior vena cava filters, 17 were not given anticoagulants (7 absolute contraindications, 10 relative contraindications), and 15 were given anticoagulants. Insertion of a percutaneous inferior vena cava filter in patients who were not given anticoagulants was followed by the development of phlegmasia cerulea dolens in four patients (24%), which was bilateral in two patients; one patient eventually died. No patients treated with inferior vena cava filter and anticoagulation had development of phlegmasia. CONCLUSIONS: Percutaneous inferior vena caval interruption effectively prevents pulmonary embolism in patients with DVT but does not impact the underlying thrombotic process and in fact may contribute to progressive thrombosis in patients who are not given anticoagulants. Anticoagulation with intravenous heparin in safe and effective therapy for DVT in most patients. We believe that percutaneous insertion of vena cava filters should not replace anticoagulation in routine proximal DVT, and those patients who require an inferior vena cava filter for failure of anticoagulation should continue to receive heparin to treat the primary thrombotic process. We caution that relative contraindications to anticoagulation should be carefully scrutinized before recommending vena cava interruption as a primary therapy for DVT. PMID- 7494364 TI - Deep vein thrombosis associated with lower extremity amputation. AB - PURPOSE: Patients undergoing lower extremity amputation are perceived to be at high risk for deep vein thrombosis (DVT). Limited data are available, however, to confirm this impression. The purpose of this study is to prospectively document the incidence of DVT complicating lower extremity amputation. METHODS: During a recent 28-month period, 72 patients (71 men, 1 woman; mean age 68 years) undergoing major lower extremity amputation (31 above-knee and 41 below-knee) were prospectively evaluated with perioperative duplex scanning for DVT. RESULTS: DVT was documented in nine (12.5%) patients (one bilateral, four ipsilateral, and four contralateral to amputation). Patients with a history of venous disease were at significantly higher risk for development of DVT (p = 0.02). Thrombi were located at or proximal to the popliteal vein in eight patients and were isolated to the tibial veins in one patient. DVT was identified before operation in six patients and after operation in three. Patients with DVT were treated with heparin anticoagulation, with no patient experiencing clinical symptoms compatible with pulmonary embolism. CONCLUSIONS: In our recent experience, lower extremity amputation is associated with DVT at or proximal to the popliteal vein in 11% of patients. Documentation of DVT prevalence is essential to assist surgeons in planning a management strategy for prevention, diagnosis, and treatment of DVT associated with lower extremity amputation. PMID- 7494365 TI - Preliminary results of a nonoperative approach to saphenofemoral junction thrombophlebitis. AB - PURPOSE: A prospective, nonrandomized study was conducted to evaluate the efficacy of a nonoperative approach of anticoagulation therapy to manage saphenofemoral junction thrombophlebitis (SFJT). METHODS: Over a 2-year period between January 1993 and January 1995, 20 consecutive patients with SFJT were entered into the study. They were hospitalized and given a full course of heparin. Duplex ultrasonography was performed before admission, both to establish the diagnosis and to evaluate the deep venous system. Two to 4 days after admission a follow-up scan was obtained to assess resolution of SFJT and to reexamine the deep venous system. Patients with SFJT alone and resolution of SFJT by duplex scanning were maintained on warfarin (Coumadin) for 6 weeks. Those with SFJT and deep venous thrombosis (DVT) were maintained on Coumadin for 6 months. Incidence of concurrent DVT and its location were noted. The efficacy of anticoagulation therapy was evaluated by measuring SFJT resolution, recurrent episodes of SFJT, and occurrence of pulmonary embolism (PE). RESULTS: There was a 40% incidence (eight of 20 patients) of concurrent DVT with SFJT. Of the eight patients, four had unilateral DVT, two had bilateral DVT, and two had development of DVT with anticoagulation. DVT was contiguous with SFJT in five patients and noncontiguous in three patients. Of 13 duplex scans obtained at 2 to 8 months follow-up, seven demonstrated partial resolution of SFJT, five had complete resolution, and one did not demonstrate resolution. There were no episodes of PE, no recurrences, and no complications of anticoagulation at maximum follow-up of 14 months. CONCLUSIONS: Anticoagulation therapy to manage SFJT was effective in achieving resolution, preventing recurrence and preventing PE within our follow up period. There is a high incidence of DVT associated with SFJT that merits careful evaluation of the deep venous system during the course of management. PMID- 7494366 TI - Risk factors for chronic venous insufficiency: a dual case-control study. AB - PURPOSE: Most epidemiologic studies on chronic venous insufficiency (CVI) are cross-sectional surveys that suggest potential risk factors by describing their population. However, these relationships could be due to the CVI population's older age. We performed a dual case-control study with multivariate analysis to address this issue. METHODS: Ninety-three patients with venous ulcers, 129 patients with varicose veins (VV), and 113 general population control patients from two hospitals were interviewed by use of a standardized questionnaire covering medical history, patient demographics, medications, and lifestyle questions. Univariate and multivariate analyses were used to compare the groups. RESULTS: Univariate analyses showed CVI to be characterized by several factors, many of which were found to be age related after multivariate analysis. Age adjusted relationships for CVI include male sex and obesity. Histories of serious leg injury or phlebitis were important associations resulting in a 2.4-fold and 25.7-fold increase in risk for CVI, respectively. After adjusting for age, subjects with VV tend to be younger and female, to more frequently have a history of phlebitis, and to report a family history of VV more frequently than control subjects. CONCLUSIONS: Many of the previously suggested associations found with CVI are in reality due to this population's greater age. Patients with CVI are older, male, obese, have a history of phlebitis, and have a history of serious leg injury. These results suggest that a prior deep vein thrombosis, either clinical or subclinical, may be a predisposing factor for CVI. PMID- 7494367 TI - Healing of venous ulcers in an ambulatory care program: the roles of chronic venous insufficiency and patient compliance. AB - PURPOSE: A nurse-managed/physician-supervised treatment program for venous ulceration was evaluated to determine the influence of venous hemodynamics, comorbidities, patient behavior, and ulcer characteristics on time to healing and time to recurrence. METHODS: The clinical course and long-term follow-up of 71 patients with 99 venous ulcers diagnosed between November 1981 and August 1994 were analyzed by a retrospective review of clinic records. Demographic data, severity of venous insufficiency, ulcer characteristics, and patient compliance were studied. Outcome variables were time to complete ulcer healing and time to first recurrence. RESULTS: Ninety-one percent of the ulcers healed completely at a median 3.4 months. There were 52 (57%) recurrences at a median 10.4 months. Ulcers on limbs with a venous refill time of 10 seconds or less demonstrated a significantly longer time to complete healing (p < or = 0.03); however, no effect on time to recurrence was observed. Patients who were in strict compliance with the treatment regimen (n = 32) had significantly faster healing (p < or = 0.02) and fewer recurrences (p < or = 0.004) compared with patients who were less compliant (n = 67). CONCLUSIONS: Most venous ulcers can be expected to heal when patients are enrolled in a nurse-managed/physician-supervised ambulatory ulcer clinic. Photoplethysmography-derived venous refill time of 10 seconds or less predicted delayed healing. Strict compliance with the treatment protocol significantly decreased the time to healing and prolonged the time to recurrence. PMID- 7494368 TI - Progress in evaluating carotid artery stenosis. PMID- 7494369 TI - Aortocaval fistula treated by aortic exclusion. AB - Aortocaval fistulas are a rare complication of spontaneous rupture of an abdominal aortic aneurysm, representing an incidence of 2% and 4%. A review of the literature revealed 159 reported cases of aortocaval fistulas. We recently had a patient admitted to our institution with an abdominal aortic aneurysm that ruptured into the vena cava. The presence of the fistula was only recognized during operation. Primary closure of the fistula was not possible, and the aneurysmal segment was excluded. To our knowledge, this is the first report of aortic exclusion being used as the surgical treatment of an aortocaval fistula. PMID- 7494370 TI - Effects of the new class III antiarrhythmic drug MS-551 and d-sotalol on canine coronary ligation-reperfusion ventricular arrhythmias. AB - The antiarrhythmic effects of a new class III antiarrhythmic agent, MS-551 [1,3 dimethyl-6-(2-[N-(2-hydroxyethyl)-3-(4-nitrophenyl) propylamino]ethylamino) 2,4(1H,3H)-pyrimidinedione hydrochloride], were investigated using canine coronary ligation-reperfusion arrhythmia models under slow and fast heart rate conditions and compared with those of d-sotalol. Slow and fast heart rate conditions were produced by using different anesthetics; i.e., halothane anesthesia for the slow heart rate condition and pentobarbital Na anesthesia for the fast heart rate condition. MS-551 prolonged QTc and suppressed the occurrence of fatal ventricular fibrillation (VF) on coronary reperfusion under either halothane or pentobarbital anesthesia. However, it also showed proarrhythmic effects, i.e., induction of torsades de pointes-like arrhythmia in 1 of 6 halothane anesthetized dogs before coronary ligation. d-Sotalol did not suppress the reperfusion VF in halothane anesthetized animals, nor did it show proarrhythmic effects. However, in the pentobarbital anesthetized animals, d sotalol suppressed reperfusion VF accompanied by proarrhythmic effects in 1 of 7 dogs. d-Sotalol did not show reverse rate dependent QT prolongation. These results indicate that although both these class III drugs have similar electrophysiological properties, such as QTc prolongation, they have different antiarrhythmic effects. Also, antifibrillatory effects of class III drugs on coronary reperfusion apparently can not be explained solely by their QT prolonging effects. PMID- 7494371 TI - Changes in gastric function and healing of chronic gastric ulcers in aged rats. AB - The present study was designed to determine the relationship between changes in gastric function caused by aging and the healing of gastric ulcers. Male Fischer 344 rats (young, 2 months; aged, 24-26 months) were used. Gastric ulcers were induced by luminal application of an acetic acid solution (60%, 0.2 ml, 45 sec). The basal and histamine-stimulated gastric acid secretions in aged animals were significantly lower than those in young animals. However, the serum gastrin levels were almost the same in the two groups. Gastric mucosal blood flow and mucosal cell proliferation were also decreased in aged animals. Gastric contractile activity induced by electrical vagal stimulation was almost the same in the two groups, while the gastric relaxative response was significantly reduced in aged animals. The development and healing rate of gastric ulcers in aged animals were almost the same as those in young animals. Histologically, regeneration of the ulcerated mucosa was significantly reduced and tissue contraction occurred in aged animals, as compared to in young animals. We concluded that aging had little or no influence on the development and healing rate of gastric ulcers in rats, but the quality of ulcer healing was quite different between young and aged animals, probably resulting from various gastric dysfunctions related to ulcer healing. PMID- 7494372 TI - Expression of alpha 2-receptor-mediated responses by insulin in primary culture of rat hepatocytes. AB - The effects of the alpha 2-adrenergic agonist, clonidine, on the glucagon stimulated glucose output from serum-free cultures of adult rat hepatocytes were examined in vitro. When hepatocytes were cultured with 10 nM dexamethasone under the serum-free condition, 1 or 10 microM clonidine did not inhibit the glucagon induced glucose production. In contrast, clonidine dose-dependently inhibited the activity concomitantly with suppression of hepatocyte cAMP production by glucagon when they were cultured with 10 nM dexamethasone and 10 nM insulin. The inhibitory effects of clonidine were completely blocked by prior treatment of hepatocytes with pertussis toxin (100 ng/ml). In addition, forskolin-stimulated cAMP production was also inhibited by alpha 2-adrenergic agonists (clonidine and oxymetazoline) in a dose-dependent manner when hepatocytes were cultured with 10 nM dexamethasone and 10 nM insulin. The inhibitory effects of alpha 2-adrenergic agonists on forskolin-stimulated cAMP production were specifically blocked when they were combined with the alpha 2-adrenergic antagonist yohimbine. Hepatocytes cultured with dexamethasone alone showed no response to the alpha 2-adrenergic agonists. The alpha 2-response was abolished when cycloheximide (0.5 microM) was added to the cultures. These results suggest that insulin develops alpha 2 adrenergic responsiveness through new protein synthesis during the primary culture of adult rat hepatocytes. PMID- 7494373 TI - The inhibitory effect of Li+ on contractile elements of intestinal smooth muscle. AB - The mechanism of the inhibitory effect of Li+ on contraction was examined in guinea pig ileal longitudinal smooth muscle. Li(+)-substitution (68.4 mM) reversed contractions induced by high K+ (45.4 mM), carbachol (1 microM) and histamine (1 microM) without changing the cytosolic Ca2+ level. Li+ also had no effect on the increase in 45Ca2+ uptake stimulated by high K+. High K+ transiently increased myosin light chain (MLC) phosphorylation, reaching a peak at 6-9 sec. Li(+)-substitution inhibited the high K(+)-induced MLC phosphorylation. In permeabilized ileal strips, contraction induced by 1 microM Ca2+ was inhibited by 10 mM Li+. The inhibitory effect was antagonized by increasing the concentration of Ca2+ or calmodulin. In the permeabilized muscle in which MLC was previously thiophosphorylated with 1 mM ATP gamma S and 3 microM Ca2+, ATP induced contraction in Ca2+ free buffer. Li+ added during this contraction did not show an inhibitory effect. In contrast, when 30 mM Li+ was added during the thiophosphorylation, the contraction induced by the subsequent addition of ATP was inhibited. Li+ (30 mM) changed neither the rate of relaxation induced by removing external Ca2+ in permeabilized muscle nor the rate of dephosphorylation of MLC induced by crude phosphatase extracted from the ileum. Li+ (15 mM), on the other hand, inhibited the rate of phosphorylation of MLC caused by crude MLC kinase extracted from the ileum. Li+ did not inhibit the calmodulin activity as measured with the (Ca2+ +Mg2+)-ATPase activity of the erythrocyte membrane. These results suggest that the inhibitory effect of Li+ on contractions is attributable to the inhibition of MLC kinase in guinea pig ileum. PMID- 7494375 TI - Participation of endogenous noradrenaline in the release of ATP by high potassium in the rat caudal artery. AB - Potassium (40 mM) evoked the release of adenine nucleotides and adenosine in rat isolated caudal arteries. This release was abolished by bunazosin. Endogenous noradrenaline was also released. Furthermore, the high potassium-induced purine release was abolished by rubbing the lumen of the vessel. These findings suggest that the high potassium releases endogenous noradrenaline, which subsequently elicits the release of ATP and related purines by acting on alpha 1-adrenoceptors on the endothelial cell. PMID- 7494374 TI - Effects of the adenosine A1-receptor antagonist on defecation, small intestinal propulsion and gastric emptying in rats. AB - We examined the effects of 1,3-dipropyl-8-cyclopentylxanthine (DPCPX) and (R)-7,8 dihydro-8-ethyl-2-(3-noradamantyl)-4-propyl-1H-imidazo[2,1 -i]purin- 5(4H)-one (KF20274), selective adenosine A1-receptor antagonists, on the gastrointestinal propulsion in rats, as compared with those of the laxative bisacodyl. DPCPX and KF20274 (p.o.) dose-dependently increased the fecal pellet output, whereas these drugs at the dose that increased defecation did not affect small intestinal propulsion or gastric emptying. Bisacodyl increased defecation and slowed gastric emptying without any influence on small intestinal propulsion. Bisacodyl, but not DPCPX or KF20274, induced diarrhea at the dose inducing defecation. The present results suggest that the adenosine A1-receptor antagonist selectively enhances the lower gastrointestinal propulsion, resulting in defecation without diarrhea. PMID- 7494377 TI - Potentiation of GABA-induced inhibition by 20-hydroxyecdysone, a neurosteroid, in cultured rat cortical neurons. AB - Effects of 20-hydroxyecdysone (20-HE), a neurosteroid, on cultured rat cortical neurons were examined using the whole cell recording technique. Under the voltage and current clamp conditions, brief application (5 sec) of 20-HE alone did not produce current changes nor any changes in the membrane potential. However, the chemical dose-dependently potentiated the GABA-induced current and hyperpolarization, which were blocked by bicuculline. These results suggest that 20-HE acts on the modulatory site of the GABAA receptor and potentiates GABAergic inhibition in rat cortical neurons. PMID- 7494376 TI - Loxiglumide, L-364,718 and L-365,260 prevent the inhibition of spontaneous acetylcholine release from the frontal cerebral cortex of freely moving rat peripherally administered with cholecystokinin-8S. AB - We examined the effect of peripheral administration of cholecystokinin (CCK)-8S on spontaneous acetylcholine (ACh) release from the frontal cortex and its prevention by loxiglumide, L-364,718 and L-365,260 in freely moving rats using intracerebral microdialysis. Subcutaneously (s.c.) administered CCK-8S at 10 and 30 micrograms/kg significantly decreased the release of ACh. The inhibitory effect of 10 micrograms/kg (s.c.) CCK-8S was prevented by loxiglumide, a mixed type of CCK-A and -B-receptor antagonist, at 1 mg/kg (intraperitoneal) and 40 micrograms/rat (intracerebroventricular, i.c.v.); L-364,718, a CCK-A-receptor antagonist, at 125 and 250 ng/rat (i.c.v.); and L-365,260, a CCK-B-receptor antagonist at 250 ng/rat (i.c.v.). These results demonstrate that peripherally administered CCK-8S inhibits spontaneous ACh release from the frontal cortex through both central CCK-A (mainly) and -B receptors. PMID- 7494378 TI - Inhibitory effect of leminoprazole on acid secretion in parietal cells isolated from guinea pig gastric mucosa. AB - Isolated guinea pig parietal cells, the function of which is similar to that of human parietal cells, were used in this study. The accumulation of 14C aminopyrine (14C-AP) was measured to study the inhibitory mechanism of leminoprazole in cells. Stimulation by 10 microM histamine, 0.1 mM carbachol, 1 microM gastrin or 1 mM db-cAMP brought about satisfactory incorporation of 14C AP, and leminoprazole concentration-dependently inhibited acid secretion induced by these stimulants. At 10(-5) M, almost 100% inhibition was observed. The IC50 values of leminoprazole obtained from its inhibitory action on histamine, carbachol, gastrin and db-cAMP-stimulated acid secretion were 4.0 x 10(-7) M, 3.5 x 10(-7) M, 2.5 x 10(-7) M and 5.6 x 10(-7) M, respectively. Thus the extent of inhibition was the same for the responses to all the secretagogues. These results indicate that the site of action of leminoprazole is intracellular and distal from cAMP (intracellular second messenger), but not at the receptor sites. The results also strongly suggest that the inhibitory action of leminoprazole on H+,K(+)-ATPase may contribute to the inhibitory effect of this drug on gastric acid secretion. PMID- 7494379 TI - Barium and strontium can substitute for calcium in stimulating nitric oxide production in the endothelium of canine coronary arteries. AB - We investigated whether Ba2+ and Sr2+ can substitute for Ca2+ in stimulating the nitric oxide (NO) production and cause relaxation in vascular smooth muscle. Ba2+ and Sr2+, like Ca2+, relaxed K(+)-depolarized canine coronary arteries in the presence of diltiazem. The Ba(2+)- and Sr(2+)-induced relaxation was endothelium dependent and was largely inhibited by NG-monomethyl-L-arginine (L-NMMA) and NG nitro-L-arginine (L-NNA), but not by indomethacin. These cations increased cyclic GMP levels in the coronary artery to a similar extent, and the increment was completely abolished by L-NMMA. The relaxation induced by each cation was attenuated in the presence of a combination of propranolol, phentolamine and atropine, and L-NNA markedly inhibited any remaining relaxation. This indicates that these cations produce endothelium-dependent relaxation through NO production as well as the relaxation mediated by neurotransmitters. The present study suggests that Ba2+ and Sr2+ can substitute for Ca2+ in the activation of the NO synthase pathway in the endothelium of canine coronary arteries. PMID- 7494380 TI - Sulpiride specifically attenuates psychological stress-induced gastric lesions in rodents. AB - Gastric lesions were developed in the communication box paradigm (CB) in mice as well as in the activity-stress paradigm (AS) in rats. Treatment with sulpiride (10-320 mg/kg, p.o.) attenuated these psychological stress-induced gastric lesions in a dose-dependent manner, while it failed to suppress those induced by physical stress such as restraint water-immersion (WI) and indomethacin treatment (IND). In contrast, treatment with famotidine (0.32-10 mg/kg, p.o.) dose dependently attenuated the gastric lesions induced by physical stress but not those by psychological stress. Pylorus-ligation study revealed that famotidine strongly reduced gastric acid secretion, whereas sulpiride minimally affected that. It was also demonstrated that physical stress (WI) enhanced acid secretion while psychological stress (CB and AS) rather depressed that. These results suggest that the mechanisms of gastric lesion formation are clearly different between physical and psychological stress and that sulpiride specifically attenuates psychological stress lesions possibly through a central mechanism. PMID- 7494381 TI - Signal transduction pathway involved in beta 3-adrenoceptor-mediated relaxation in guinea pig taenia caecum. AB - Experiments were carried out to examine the components of the intracellular second messenger system that is involved in beta 3-adrenoceptor (atypical beta adrenoceptors)-mediated relaxation in the guinea pig taenia caecum. Propranolol and butoxamine caused competitive antagonism of the relaxant response to isoprenaline. However, propranolol or butoxamine did not significantly affect the relaxant responses to CGP 12177 (4-[3-[(1,1-dimethylethyl)amino]-2 hydroxypropoxy]-1,3-dihydro-2H- benzimidazol-2-one), a beta 3-adrenoceptor agonist. The concentration-response curves of the isoprenaline-induced increase in adenosine 3',5'-cyclic monophosphate (cyclic AMP) levels were shifted to the right in a parallel manner by propranolol and butoxamine. However, propranolol or butoxamine did not significantly affect the concentration-response curve for the CGP 12177-induced increase in cyclic AMP levels. MDL 12330 (cis-N-(2 phenylcyclopentyl)-azacyclotridec-1-en-2-amine) inhibited the isoprenaline- or CGP 12177-induced increase in cyclic AMP levels. These results suggest that the production of cyclic AMP contributes to the beta 3-adrenoceptor (or atypical beta adrenoceptor)-mediated relaxation of the guinea pig taenia caecum. PMID- 7494382 TI - Effects of NIP-502 on antigen-induced bronchial responses and allergic reactions in animal models. AB - We examined the effect of a newly synthesized pyridazinone derivative, NIP-502 [4 chloro-5-(3-ethoxy)-4-phenoxybenzamine)-3(2H)-pyridazinone], on antigen-induced bronchial responses and allergic reactions in several animal models. NIP-502 (10 mg/kg, p.o.) inhibited the antigen-induced immediate asthmatic response in passively sensitized guinea pigs. The inhibitory effect was also observed in metyrapone (an inhibitor of 11 beta-hydroxylase)-pretreated guinea pigs. NIP-502 improved ovalbumin (OA)-induced airway hyperresponsiveness to acetylcholine and inhibited the OA-induced increase in the number of inflammatory leukocytes in the bronchoalveolar lavage fluid. These inhibitory effects on OA-induced responses were similar to those of prednisolone. NIP-502 also showed an inhibitory effect on the passive cutaneous anaphylactic reaction in rats but did not inhibit the reversed cutaneous anaphylactic reaction, reversed Arthus reaction or delayed type hypersensitivity reaction. On the other hand, prednisolone showed broad inhibitory effects except for the reversed cutaneous anaphylactic reaction. In the in vitro study, NIP-502 (30 microM) significantly inhibited Formyl-Met-Leu Phe-induced superoxide anion production by the guinea pig alveolar macrophages. These results indicate that the inhibitory effects of NIP-502 on bronchial responses are similar to those of prednisolone, but this compound seemed to act more selectively on the respiratory tract than prednisolone. Because of its effectiveness against a variety of bronchial responses, NIP-502 may be useful in the treatment of bronchial asthma. PMID- 7494383 TI - Profiles of the antinociceptive effect of R-84760, a selective kappa-opioid receptor agonist, in the formalin test in mice. AB - The antinociceptive effect of a selective kappa-opioid receptor agonist R-84760, (3R)-3-(1-pyrrolidinylmethyl)-4-[(1S)-5,6-dichloro-1-indancarbo nyl]- tetrahydro 1,4-thiazine hydrochloride, in the second phase of the formalin test, a model of tonic pain, was examined in mice. R-84760 had a 2700 times more potent antinociceptive effect than morphine. The effect of R-84760 was antagonized by subcutaneously administered nor-binaltorphimine, a kappa-selective opioid receptor antagonist. Both intracerebroventricularly and intrathecally administered nor-binaltorphimine partially antagonized the antinociceptive effect of R-84760. Intrathecally administered phentolamine, an alpha-adrenoceptor antagonist, attenuated and desipramine, a noradrenaline reuptake inhibitor, augmented the antinociceptive effect of R-84760. Intrathecally administered noradrenaline attenuated the nociceptive response in the second phase of the formalin test. Intrathecally administered (+/-)-3-(2-carboxypiperazin-4-yl) propyl-1-phosphonic acid (CPP), an N-methyl-D-aspartate (NMDA)-receptor antagonist, reduced and threo-beta-hydroxyaspartate, a reuptake inhibitor of glutamate, augmented the second phase nociceptive response. However, R-84760 did not influence the intrathecally injected NMDA-induced nociceptive response. These results suggest the following: R-84760 produces an extremely potent antinociceptive effect against tonic pain through the kappa-opioid receptors; the sites of action of subcutaneously administered R-84760 are the supraspinal and spinal loci in the central nervous system; and a part of the mechanism of the antinociceptive effect of R-84760 is activation of the descending noradrenergic pathway. PMID- 7494384 TI - 5-HT1A-receptor subtype mediates the effect of fluvoxamine, a selective serotonin reuptake inhibitor, on marble-burying behavior in mice. AB - The effect of fluvoxamine, a selective serotonin (5-HT) reuptake inhibitor, was studied in a model of anxiety and/or obsessive compulsive disorder (OCD) in mice. In the anxiety/OCD model, marble-burying behavior, marble-burying was significantly suppressed by fluvoxamine at 30 and 60 mg/kg, p.o. and the monoamine reuptake inhibitor clomipramine, at 60 mg/kg, p.o. No suppressive effect, however, was observed by the selective norepinephrine reuptake inhibitor desipramine at doses from 15 to 60 mg/kg, p.o. Suppressive effects were obtained by the serotonergic anxiolytic buspirone at 30 and 60 mg/kg, p.o. and the benzodiazepine anxiolytic diazepam at 10 mg/kg, p.o. The effect of fluvoxamine on marble-burying was slightly attenuated after repeated administration. On the other hand, both the effects of buspirone and diazepam completely disappeared after repeated administration. Effect of fluvoxamine on the marble-burying was unaffected by the 5-HT2 antagonist ritanserin. However, the 5-HT1A antagonist NAN 190 (1-(2-methoxyphenyl)-4-[4-(2-phthalimido)butyl] piperazine) inhibited the suppressive effect of fluvoxamine on the marble-burying. From these results, the 5-HT1A-receptor subtype may be involved in the suppressive effect of fluvoxamine on the marble-burying, but the 5-HT2-receptor subtype is not involved in this effect. PMID- 7494385 TI - Neither the 5-HT1A- nor the 5-HT2-receptor subtype mediates the effect of fluvoxamine, a selective serotonin reuptake inhibitor, on forced-swimming-induced immobility in mice. AB - The effect of fluvoxamine, a selective serotonin (5-HT) reuptake inhibitor, was studied in the forced-swimming test, a model of depression, in mice. Fluvoxamine at 60 mg/kg, p.o. significantly decreased the immobility time in the forced swimming test. A similar effect was observed by the selective norepinephrine reuptake inhibitor desipramine at the same dose. Furthermore, the suppression of immobility time was slightly potentiated by repeated administration of fluvoxamine, and a significant effect was observed at 30 mg/kg, p.o. The effect of fluvoxamine on forced-swimming was unaffected by the 5-HT2 antagonist ritanserin. On the other hand, the 5-HT1A antagonist NAN-190 (1-(2-methoxyphenyl) 4-[4-(2-phthalimido)butyl] piperazine) potentiated the effect of fluvoxamine on forced-swimming. It is expected, however, that a 5-HT1A antagonist should antagonize the effect of fluvoxamine when 5-HT1A mediates the suppressive effect of fluvoxamine on the immobility time in forced-swimming. From these results, neither the 5-HT1A- nor the 5-HT2-receptor subtype is involved in the suppressive effect of fluvoxamine on the immobility associated with forced-swimming. PMID- 7494386 TI - Change in membrane permeability induced by amyloid beta-protein fragment 25-35 in brain neurons dissociated from rats. AB - Effects of amyloid beta-protein fragment 25-35, A beta P(25-35), on the membrane permeability of organic molecules were examined in the brain neurons dissociated from rats by using an argon laser (equipped in flow cytometer and laser microscope) and a combination of two fluorescent dyes, fluo-3-AM and ethidium bromide. A beta P(25-35) at concentrations of 1 microM or greater induced both leakage of fluo-3 from the neurons and permeation of ethidium across the membrane in a dose-dependent manner, although both dyes are highly impermeant to the intact plasma membrane. Thus, A beta P(25-35) seems to increase not only membrane permeability of inorganic ions such as Ca2+, Na+ and K+, as previously suggested, but also that of organic molecules. Therefore, the brain neuron membrane is suggested to lose its integrity in the presence of A beta P(25-35) that leads to neuronal death. PMID- 7494387 TI - Involvement of raphe-hippocampal serotonergic and septo-hippocampal cholinergic mechanisms in the penile erection induced by FR121196, a putative cognitive enhancer. AB - FR121196 (N-[4-acetyl-1-piperazinyl]-4-fluorobenzenesulfonamide), a putative cognitive enhancer, induced penile erection in naive rats; the dose-response curve was bell-shaped with the maximum response obtained at the dose of 3.2 mg/kg. The response to FR121196 was abolished in rats treated with intra-raphe injections of 5,7-dihydroxytryptamine or systemic injections of p chlorphenylalanine (150 mg/kg, i.p. for three consecutive days) as well as in rats with electrolytic medial-septum lesion or surgical fimbria-fornix lesion. In addition, the penile erection induced by FR121196 (3.2 mg/kg) was dose dependently attenuated by pindolol (0.1-3.2 mg/kg), a serotonin (5-HT)1 antagonist with beta-antagonistic activity, but not by metoprolol, a selective beta=antagonist. The inhibitory activity was shared by ICS205-930, a 5-HT3 antagonist, but not by ketanserin, a 5-HT2 antagonist, or sulpiride, a dopamine D2 antagonist. Scopolamine (0.032-1 mg/kg), but not methyl-scopolamine (0.032-1 mg/kg), also attenuated the penile erection induced by FR121196. Neurochemical analysis revealed that intraperitoneal injection of FR121196 significantly elevated the levels of 5-HT and its major metabolite 5-hydroxyindoleacetic acid (5-HIAA) in the hippocampus and that raphe-lesion significantly reduced both 5-HT and 5-HIAA levels without affecting choline-acetyltransferase activity in all cortical and subcortical regions examined. It is thus postulated that FR121196 facilitates the raphe-hippocampal serotonergic pathway resulting in an activation of the septo-hippocampal cholinergic pathway and finally induces the penile erectile response. PMID- 7494389 TI - Nursing informatics: update on the newest nursing specialty. PMID- 7494388 TI - Antiarrhythmic effects of bisaramil on triggered arrhythmias produced by intracoronary injection of digitalis and adrenaline in the dog. AB - Antiarrhythmic effects of bisaramil were examined by using new in vivo triggered arrhythmia models, and they were compared with those of other antiarrhythmic drugs. Bisaramil (3-10 micrograms, i.c.) suppressed triggered ventricular arrhythmias that were produced during pauses between trains of rapid ventricular stimulation (cycle length: 250 msec, train number: 15) in anesthetized open-chest dog hearts administered with subtoxic doses of digitalis or adrenaline to the anterior descending coronary artery. The potencies of bisaramil, disopyramide, lidocaine and flecainide suppressing digitalis-induced triggered ventricular arrhythmias were similar to those suppressing adrenaline-induced ones. The potency of verapamil for suppressing digitalis-induced triggered ventricular arrhythmias were weaker than that for suppressing the adrenaline-induced ones. Bisaramil was the most effective among the antiarrhythmic drugs used in the present experiment. Since bisaramil has been reported to be effective in suppressing other canine automatic ventricular arrhythmias, and the triggered ventricular arrhythmias occur in clinical situations, bisaramil may become a useful drug for the treatment of clinical arrhythmias. PMID- 7494391 TI - Communication technology in rural areas: an educator's perspective. PMID- 7494390 TI - Communication in rural health care. PMID- 7494392 TI - The advanced registered nurse practitioner in rural practice. AB - Availability of health care to rural areas is limited. FNP's in rural practice have the ability to address rural needs and tailor interventions to the specific attributes of rural populations. The objective of this article is to define the role and the functions of the FNP and to support and defend this role in the rural setting. Much of the FNP research presented in the literature to date focuses on urban populations; therefore, the unique health care problems and needs of rural populations will first be defined as they relate to the FNP's scope of practice. PMID- 7494393 TI - Why Kansas needs prescriptive authority for ARNP's. PMID- 7494394 TI - Advanced registered nurse practitioners: preparation, roles and practice settings. AB - Two presidents have focused on the goal of providing access to health care for every U.S. citizen (DeAngelis, 1994). Advanced registered nurse practitioners (ARNPs) can help with the accomplishment of this goal through collaboration and participation as a multidisciplinary team member. "Health care problems and systems are often so complex, one person can no longer know everything about anything" (Keiser, 1994, p. 328). PMID- 7494395 TI - Prescriptive authority for nurse practitioners in Kansas. PMID- 7494396 TI - Tyrosinaemia--treatment and outcome. AB - Tyrosinaemia type I is, untreated, a fatal disease: in the acute form from liver failure, in the chronic form often from hepatocellular carcinoma. Acute neurological crisis is also a cause of death. Traditionally the treatment has been with diet, but for a decade liver transplantation has been the ultimate treatment. The continuous production of the pathological metabolites in the kidneys after transplantation appears to be without significance. Introduction of the enzyme inhibitor NTBC in the treatment of tyrosinaemia has reduced the need for liver transplants. Neonatal screening may be justified as efficient treatment has become available. The complex phenotype of lethal albino mice, with severe alterations in gene expression, has been shown to be caused by fumarylacetoacetase deficiency. Prolonged hypoglycaemia in otherwise adequately treated tyrosinaemia patients may result from depressed expression of genes coding for enzymes in gluconeogenesis, as seen in the mouse model. Self-induced genetic correction in liver tissue that occurs in many tyrosinaemia patients may reduce the risk of liver failure in some patients. PMID- 7494397 TI - Cystinosis. AB - Nephropathic cystinosis is an autosomal recessive inborn error of metabolism characterized by the lysosomal storage of the disulphide amino acid cystine. It produces a variety of clinical manifestations including failure to thrive, the renal Fanconi syndrome, eye findings, and end-stage renal disease. A variety of phenotypes are known; however, the molecular defect underlying any of the forms has not yet been identified. Therapy of cystinosis with cysteamine averts the otherwise inevitable renal failure, but systemic therapy does not improve the corneal keratopathy. A number of presentations in this review detail approaches to gene identification, systemic therapy with cysteamine, measurement of cystine, and pathophysiological effects at the cellular and clinical level. PMID- 7494398 TI - Recent advances in the treatment of cystinosis. AB - Cysteamine bitartrate capsules (Cystagon) have been approved by the US Food and Drug Administration for use in patients with nephropathic cystinosis. Plasma cysteamine concentrations were virtually identical at various times following ingestion of either cysteamine hydrochloride or Cystagon capsules in 24 normal control subjects. A transfer study was done with eight cystinosis patients who had been receiving either cysteamine hydrochloride or phosphocysteamine for many years. The plasma cysteamine concentration was significantly higher 2h after Cystagon and the leukocyte cystine content was significantly lower at all times after Cystagon compared to older forms of the drug. These differences are probably the result of greater patient compliance in taking the capsules compared to the older, liquid forms of the drug. A new method for following the course of renal glomerular deterioration in diseases such as cystinosis has been published recently. This method was used to re-analyse data on the efficacy of cysteamine treatment and to re-analyse new data on treating cystinosis patients with either of two doses of cysteamine (1.30 g/m2 per day and 1.95 g/m2 per day). This new method agrees well with other methods and shows that both doses of drug are equally effective in maintaining glomerular function. PMID- 7494399 TI - The future for treatment by bone marrow transplantation for adrenoleukodystrophy, metachromatic leukodystrophy, globoid cell leukodystrophy and Hurler syndrome. AB - Within the past decade, bone marrow transplantation has been applied to over 200 patients worldwide with the intention of treating storage diseases. Bone marrow transplantation has provided a method for treatment of adrenoleukodystrophy, metachromatic leukodystrophy, globoid cell leukodystrophy and Hurler syndrome. After engraftment, significant improvement in the clinical course of each of these diseases occurs. Survival data of engrafted patients are superior to those of non-transplanted. Engraftment and the resulting enzymatic reconstitution are concordant. Outcomes based on neuropsychological tests indicate continued maintenance and in some cases increase in cognitive function. Magnetic resonance imaging as well as spectroscopic examinations of the brain provide further evidence that positive changes occur in the central nervous system following long term engraftment. A better quality of life follows engraftment. Greater gains from use of bone marrow transplantation for these particular storage diseases will occur in the future. Earlier diagnosis will allow bone marrow transplantation in the presymptomatic stage at a younger age, providing an enhancement of positive effects noted from such treatment. At the same time, advances in bone marrow technology will serve to reduce the risk factors involved with the bone marrow transplantation process itself. These two factors taken together will be more than additive in providing benefits from use of bone marrow transplantation. PMID- 7494400 TI - Neuropsychological outcomes of several storage diseases with and without bone marrow transplantation. AB - Neuropsychological assessment is essential in providing documentation of the untreated natural history of storage diseases associated with dementia and quantifying the effectiveness of treatment on central nervous system function. Baseline characterization and outcome of bone marrow transplantation (BMT) for three leukodystrophies and three mucopolysaccharidoses are presented. Results suggests that BMT for Hurler syndrome, adrenoleukodystrophy, and globoid cell leukodystrophy can be effective in preventing dementia if done early enough in the disease. Sanfilippo and Hunter syndromes do not benefit and BMT is not recommended. For metachromatic leukodystrophy, BMT is not recommended for symptomatic early-onset forms of the disease. Further longitudinal follow-up is needed to determine whether the benefits outweigh the risks of BMT for late-onset and preclinical metachromatic leukodystrophy. PMID- 7494401 TI - The management and outcome of propionic and methylmalonic acidaemia. AB - The management of the severe variants of methylmalonic and propionic acidaemia remains difficult. With conventional therapy of diet, carnitine and antibiotics, mortality is high and long-term complications are common. Liver transplantation appears to be an important alternative. PMID- 7494402 TI - Komrower Lecture. Adrenoleukodystrophy: natural history, treatment and outcome. AB - Our laboratory has identified nearly 2000 patients with X-linked adrenoleukodystrophy (ALD) and conducted therapeutic trials in groups of patients who represent the major phenotypes. We report recent results of dietary therapy with a mixture of glyceryl trioleate and glyceryl trierucate oil, also referred to as Lorenzo's Oil, in the asymptomatic and childhood cerebral phenotypes. Fifty three patients started this therapy at a mean age of 7.5 years at a time when they were free of neurological symptoms. Although analysis of data is hampered by the lack of a concurrent control group, follow-up studies after 39 months of therapy suggest that subsequent neurological involvement was less frequent and less severe than anticipated from historical controls. Retrospective analysis of the effect of the oil in patients with the severe childhood cerebral phenotype indicates that there was a slight but statistically significant slowing of clinical progression and delay of death. PMID- 7494403 TI - Liver transplantation in two cases of propionic acidaemia. AB - Orthotopic liver transplantation (OLT) was performed in two patients with propionic acidaemia, a 7-year-old boy and a 9-year-old girl, diagnosed with a severe neonatal form with high risk of metabolic decompensation. In both cases the metabolic liver functions recovered within the 12 postoperative hours; no clinical symptoms of propionic acid toxicity, metabolic acidosis, severe hyperammonaemia, hyperglycinaemia or haematological abnormalities were observed. In both cases insulin-dependent diabetes mellitus occurred early after OLT (persisting in the boy's case). Severe post-transplantation complications were observed (acute rejection and CMV infection in both patients) which did not trigger metabolic decompensation. The boy developed chronic rejection and vanishing bile duct syndrome due to incomplete hepatic arterial thrombosis. He required permanent in-patient care with chronic hyperammonaemia and neurological sequelae involving the basal ganglia and died 15 months after OLT. The girl left hospital after 2 months and is presently leading a normal life with almost no dietary protein restriction (40 g protein per day). Urinary urea excretion and daily protein intake increased after liver transplantation. Propionyl- and tiglylglycine disappeared immediately after OLT. Urinary methylcitrate and 3 hydroxypropionate remained at concentrations corresponding to those before OLT. However, the total of all characteristic metabolites of organic acid analysis was reduced to 50-60% of the values before OLT in both patients. Propionylcarnitine was still detected at significant concentrations. Plasma odd-chain fatty acid concentrations decreased continuously after OLT only in the girl's case. Tissue of both transplanted livers showed increased odd-chain fatty acid concentrations 9 and 15 months after OLT, respectively, in both patients. We consider that at present OLT should only be performed in severe forms of propionic acidaemia. PMID- 7494404 TI - The hepatic glycogen storage diseases--problems beyond childhood. AB - The introduction of continuous nocturnal enteral glucose feeds and uncooked cornstarch has improved the prognosis for patients with the hepatic glycogen storage diseases. An increasing number of patients are surviving into adulthood in better health, but still at some medical cost. In this review we examine bone mineralization, renal function, hepatic tumours, and vascular endothelial function in GSD I and cardiac function in GSD III. All females over the age of 5 years with GSD I, III, VI and IX had morphologically polycystic ovaries. Thirteen adult GSD I patients have been studied, and been found to have poor bone mineralization and marked renal glomerular and tubular dysfunction. More than half of these patients also had focal hepatic lesions on sonography and yet vascular endothelial function was preserved in the face of hyperlipidaemia. In 12 GSD III patients, one had a focal hepatic lesion and 6 had pronounced left ventricular hypertrophy, although cardiorespiratory function was normal. These data emphasize the multisystem nature of these disorders and highlight the need for careful longterm follow-up. PMID- 7494406 TI - Cystic fibrosis: five years beyond the gene. PMID- 7494405 TI - Disorders of mitochondrial long-chain fatty acid oxidation. AB - The oxidation of long-chain fatty acids requires a series of enzymes which are located in or on the mitochondrial membranes. These include carnitine palmitoyltransferases I and II, a carnitine-acylcarnitine translocase and, newly discovered, very long-chain acyl-CoA dehydrogenase and the mitochondrial trifunctional protein. These last two chain-shorten acyl-CoA esters to the point where they can be transferred to the more soluble medium- and short-chain specific enzymes within the mitochondrial matrix. The disorders of long-chain fatty acid oxidation show a rather similar range of clinical and biochemical features, though with different emphasis in the different conditions. Patients with severe defects usually present early with acute attacks of hypoketotic hypoglycaemia and impaired liver function, or with cardiomyopathy or cardiac arrhythmia. In milder variants, skeletal myopathy with intermittent myoglobinuria develops later in life. 3-Hydroxyacyl-CoA dehydrogenase deficiency is unusual in producing peripheral neuropathy and retinitis pigmentosa. Treatment is based on the avoidance of fasting and replacement of normal dietary fat by medium-chain triglyceride, the medium-chain fatty acids entering the mitochondria in a carnitine-independent manner and bypassing the long-chain part of the spiral. Diagnosis must ultimately be based on direct assay of the enzyme involved, but preliminary indicators may come from determination of carnitine and intermediate metabolites in plasma, urinary organic acid profiling, and radioisotopic screening assays with lymphocytes or cultured fibroblasts. PMID- 7494407 TI - Development of mouse models for cystic fibrosis. AB - Using gene targeting in embryonal stem cells it is now possible to create accurate genetic models of inherited human disease in the mouse. The value of an animal model of cystic fibrosis is in providing clarification of disease pathogenesis, genotype-phenotype correlation, the identification of other relevant genetic factors, and as the optimal test system for novel therapeutic intervention. Correction of the basic defect by a somatic gene therapy approach is an attractive approach to disease treatment. CF mouse models have been described which display the characteristic electrophysiological defect and thus both safety and efficacy can be monitored in these animals. Modulation of Cftr levels in transgenic animals and the results on disease phenotype give some indication of the level of gene expression necessary to give clinical effect. PMID- 7494408 TI - Gene therapy for cystic fibrosis. AB - Recent studies have identified the underlying molecular defect in cystic fibrosis (CF). Reduced or absent cAMP-mediated chloride transport in epithelial-lined organs characterizes this disease. With the identification of the CF gene, gene therapy has become a potential novel form of treatment for this disease. This article reviews the rapid progress in CF research from the understanding of the bioelectric defect to the recently begun human gene therapy trials. PMID- 7494410 TI - Designing trials for gene therapy. PMID- 7494409 TI - Treatment of cystic fibrosis based on understanding CFTR. AB - In the five years since the gene associated with cystic fibrosis was isolated, rapid progress has been made in understanding the structure and function of CFTR, the gene product. Based on the knowledge that CFTR is a cAMP-regulated chloride channel in the apical surface of epithelia in many tissues, new approaches to treating CF disease have emerged. These include development of agents to open alternative chloride channels, to relocate mutant CFTR, which commonly is mislocalized because of a trafficking defect, and to deliver CFTR using protein or gene therapy. Although the underlying basis of each of these approaches is now well understood, major practical problems face each of them. Nevertheless, it is difficult not to be optimistic that new therapies will emerge. PMID- 7494411 TI - Carrier screening in the community. AB - A number of different models of cystic fibrosis (CF) carrier screening have now been tested in pilot trials. Apart from opportunistic and cascade testing (which are strictly speaking not true forms of screening), the major programmes have been directed either to young adults in primary care or to pregnant women in antenatal clinics. Only in the latter form of screening has sufficient data been collected to allow conclusions to be reached on the optimum mode of delivery. It seems very probable that when CF carrier screening passes into routine service, it will be the antenatal couple model that is used. PMID- 7494412 TI - Where are all the nurses? PMID- 7494413 TI - Designing nursing's future: will you be a part? PMID- 7494414 TI - Cultural diversity: considerations for ethical and culturally sensitive practice. PMID- 7494415 TI - Position statement on cultural diversity in nursing practice. ANA (American Nurses' Association) Council on Cultural Diversity in Nursing Practice, Congress of Nursing Practice, in 1991. The Maryland Nurses Associations, 1994. PMID- 7494416 TI - Are you committing strategic resume errors that can sabotage your job-search campaign? PMID- 7494417 TI - Tax on nursing? Maryland Association says no, again. PMID- 7494418 TI - Messenger more important than message in HIV prevention. PMID- 7494420 TI - Your references: important allies in your job campaign. PMID- 7494419 TI - Maryland's nursing school pins. PMID- 7494421 TI - Ethical dilemmas: nurses in clinical practice as the patient advocate. Understanding the ethics or patient care. PMID- 7494422 TI - Do not resuscitate orders: the nurses role. PMID- 7494423 TI - Position statement on nursing care and do-not-resuscitate decisions. ANA (American Nurses' Association) Task Force on the Nurses Role in End of Life Decisions. PMID- 7494424 TI - Networking your way to a new job? Realize its limitations. PMID- 7494425 TI - Legal perspectives. Medical malpractice defenses. PMID- 7494426 TI - Accessing patient care advisory committees: preparing the clinical nurse as a patient advocate in this forum. PMID- 7494428 TI - Mail vs telephone--how should you initiate contact with your targeted employers? PMID- 7494427 TI - Restructuring nursing: a concern of the Maryland Nurses Association. PMID- 7494429 TI - Legal perspectives. Economic malpractice. PMID- 7494430 TI - Legal perspectives. Provider-patient relationship. PMID- 7494431 TI - Human relations weaknesses--a leading cause of unsuccessful job campaigns. PMID- 7494432 TI - Legal perspectives. Negligence theory. PMID- 7494433 TI - Making the decision to change career. PMID- 7494434 TI - What if a patient requests assisted suicide? PMID- 7494435 TI - [Alcohol consumption in schoolchildren]. AB - BACKGROUND: The onset of alcohol consumption begins very early in the Spanish society; generally, at the school age. The target of this study is to identify the factors that are linked to alcohol use and abuse, at this age. METHODS: A survey was designed and carried out among 1,816 students of 6 schools of Barcelona and Lleida, Spain (97.5% response rate) between 13 and 18 years old included, through a questionnaire filled-in in the classroom. RESULTS: 85.5% had ever consumed alcoholic drinks, 7.3% reported to drink alcohol every day and 39.6% said that they had drunk four or more glasses of an alcoholic beverage at least once. The multivariate analysis showed that smoking habitually, with an odds ratio (OR) of 9.7; having a larger amount of own money to buy alcohol (OR = 1.5); and the expectations that alcohol helps to enjoy and get along with friends (OR = 2.4) were the most important factors identified as associated with problematic alcohol consumption. CONCLUSIONS: The study points to several factors related to the habit of smoking, the influence of the environment of schoolchildren and the expectations associated to alcohol consumption are related with the consumption of alcoholic drinks, especially from the beginning in the high school. PMID- 7494436 TI - [Measles seroprevalence in the schoolchildren of Valencia. The Measles Study Group]. AB - BACKGROUND: People born between 1978 and 1982 were the most affected by measles in the outbreaks occurring in the Valencian Community (Spain) in 1993. This age group has not been systematically immunized against measles and has not suffered the last large measles epidemics. HYPOTHESIS: global seroprevalence against measles in this age group is inadequate to prevent new outbreaks. METHODS: Prospective seroepidemiologic study in a sample of school-attending children, born between 1978 and 1982. Randomized sampling of all classrooms 5th to 8th grade high school of a Health Area; stratification depending on the population of the village (< 5,000, 5,001-20,000, > 20,000 inhabitants). Antecedents of measles immunization and the disease were collected. Measles IgG antibodies were measured by enzyme immunoassay, antibody titers 1:80 or higher were considered protective. RESULTS: Sample of 410 subjects, belonging to 17 classes. Measles antibody prevalence was 80.2% (95% CI: 76.6-83.8%), and was higher in less populated villages (p < 0.03). Immunization status was known in 253 subjects (61.7%). Protective titers were present in 86.1% of the vaccinated and in 68.1% of the non vaccinated (p < 0.001). CONCLUSIONS: There is a large percentage of subjects born between 1978 and 1982 unprotected against measles. Elimination of the disease will not be able unless an extraordinary vaccination campaign to include these subjects is taken. PMID- 7494437 TI - [An evaluation of the clinical competence of a population of specialist physicians educated by the medical internship and residency system]. AB - BACKGROUND: The quality of physicians who have undergone resident official training (MIR) should logically be better than that of the remaining physicians who were not able to enter into this official training. The present study was designed with the aim of verifying this hypothesis. METHODS: A sample of physicians who underwent the MIR examination in 1982 and who upon passing the same were permitted to initiate the MIR training in 1983 was selected. The group was subdivided into MIR and no MIR and according to the specialty followed. When the physicians were practicing as specialists two types of surveys were carried out with one being by telephone and the other personal in which the personal characteristics, preparation for the MIR test, professional satisfaction and personal motivation were analyzed. The pharmaceutic prescriptions of both groups were analyzed according to indicators of the Servei Catala de la Salut (Catalonian Health Service) and the opinion of colleagues of each of the members of each group was evaluated with another questionnaire. The written resolution of hypothetical clinical cases were given to each of the individuals included. A level of global competence defined as a percentage for the following components was identified using: curricular evaluation (10%), professional satisfaction (20%), personal motivation (10%), hypothetical case resolution (35%) and peer opinion (25%). RESULTS: The global competence of the physicians trained under the MIR system was greater than that of the no MIR group (p < 0.01). On analysis by sections the differences of greatest note were observed in the resolution of hypothetical cases (p < 0.0001), curricular evaluation (p < 0.0001) and the quality of pharmaceutical prescription (p < 0.0001). The differences were less of note in comparison of personal motivation (p < 0.02) and professional satisfaction (p < 0.02). No differences were observed in peer opinion. CONCLUSIONS: The professional quality of physicians trained by MIR who presented for the 1982 examination in Catalonia and practiced in medical specialties is greater than that of a comparable group with regard to professional practice. PMID- 7494438 TI - [Hyperlipidemia and nephropathy: cause or consequence?]. PMID- 7494439 TI - [HIV-1 group O]. PMID- 7494440 TI - [A comparative study of the clinical and histological characteristics between classic nemaline myopathy and that associated with the human immunodeficiency virus]. AB - BACKGROUND: Several cases of nemaline myopathy (NM) have been described in patients with human immunodeficiency virus (HIV) infection. The clinical and histological characteristics of classical NM are known, but remain to be defined in HIV positive patients. METHODS: A retrospective review of the NM over a 12 year period (1982-1993) was carried out. The clinical and histological data of the classical NM were compared with those associated to the HIV. RESULTS: Eight cases of NM were collected: five in HIV negative patients (classical NM) and three in HIV positive patients. The latter patients with NM were all males with no family history of myopathy. Motor development had been normal and was not associated with disorders in other systems and the muscular symptoms were slight. On muscle biopsy a lower percentage of fibers with nemaline bodies and type 2 muscle fiber atrophy were observed, as well as slight alterations in the oxidative reactions. A second muscle biopsy performed two years after in one of these patients was normal. Females predominated in the group of HIV negative patients with family history, motor retardation during infancy and different extramuscular alterations being frequent. The muscle symptoms were more marked and severe in the two cases with respiratory musculature involvement. Histologic study demonstrated a higher percentage of nemaline bodies, predominance of type 1 muscle fiber atrophy and frequent alterations in the oxidative reactions. CONCLUSION: Nemaline myopathy presented in patients with HIV infection differs from the classically described form. The presence of nemaline bodies on muscle biopsy in HIV positive patients may be interpreted more as an epiphenomenon in relation with HIV infection than as a primary myopathy. PMID- 7494441 TI - [Third list of words with misleading translation in medical English]. PMID- 7494442 TI - [The impact of human immunodeficiency virus infection on mortality in youth in Spain]. PMID- 7494443 TI - [Medico-clinical audit and the quality of care]. PMID- 7494444 TI - [A call to attention on the organization of palliative care in oncology]. PMID- 7494445 TI - [The osteoarticular manifestations associated with parvovirus B19 infection in the course of an infectious outbreak]. PMID- 7494446 TI - [The epidemiology of tuberculosis in El Ferrol]. PMID- 7494447 TI - Developmental expression of parvalbumin mRNA in the cerebral cortex and hippocampus of the rat. AB - Parvalbumin (PARV) belongs to the family of calcium-binding proteins bearing the EF hand domain. Immunocytochemical studies in the cerebral cortex have demonstrated that neurons containing PARV include two types of GABAergic interneurons, namely, basket and axo-axonic chandelier cells. The present study examines the onset and pattern of PARV mRNA expression during the development of rat neocortex and hippocampus by means of 'in situ' hybridization with an oligonucleotide probe corresponding to rat PARV cDNA. In animals aged P0-P6 no signal was detected above background in neocortex or hippocampus. At P8, a few cortical cells displayed a number of silver grains just above background levels. By P10 PARV mRNA-expressing cells in the neocortex were detected almost exclusively in layer V of somatosensory, frontal and cingulate cortices. At P12 PARV mRNA was mainly detected in layers IV, V and VIa. By P14 there was a marked overall increase in the entire neocortex, including layer II-III, both in the number of cells and in their intensity of labelling. Further maturation in the pattern of PARV mRNA concentration was observed between P16 and P21. In the hippocampus low hybridization was observed at P10-P12. In subsequent stages both the number of positive cells and the intensity of labelling increased steadily. No clear-cut radial gradients for the expression of PARV mRNA were observed in the hippocampal region. Our results show that the developmental radial gradient followed by PARV mRNA expression in the neocortex does not follow an 'inside-out' gradient, consistent with previous immunocytochemical findings. Taken together, these data indicate that the developmental sequence followed by the PARV protein directly reflects mRNA abundance and suggest that PARV mRNA expression correlates with the functional maturation of cortical interneurons. PMID- 7494449 TI - Apoptotic DNA fragmentation in the rat cerebral cortex induced by permanent middle cerebral artery occlusion. AB - Recent investigations have demonstrated internucleosomal DNA fragmentation in ischemic neuronal tissue. This type of fragmentation is characteristic of programmed cell death or apoptosis and suggests that neuronal death in stroke may be more complex than simple necrotic death. The present experiments provide a detailed examination of the regional localization and time course for apoptotic DNA fragmentation in the cerebral cortex following focal cerebral ischemia. Spontaneously hypertensive rats were subjected to permanent right middle cerebral artery occlusion and the cerebral cortices were examined for evidence of DNA fragmentation using electrophoretic, flow cytometric, and histological approaches. An electrophoretic examination of cortical DNA at 24 h after the occlusion indicated that the majority of nucleosomal ladders were in the transition zone or penumbra and the core of the infarction, with no fragmentation apparent in the contralateral normal cortex. A flow cytometric analysis of DNA fragmentation in intact cells revealed a similar pattern, with increased fragmentation observed in ischemic cortex vs. the contralateral cortex. Saggital sections taken 1.5 mm lateral to midline were collected from animals at 1, 4, and 24 h after the infarction and DNA fragmentation was examined histologically by terminal deoxynucleotidyl transferase mediated dUTP-biotin nick end labeling (TUNEL) staining. Quantitative analysis of these sections indicated that DNA fragmentation can be observed in the anterior and central area of the infarctions as soon as 1 h after the occlusion and that the extent and magnitude of the fragmentation increases at 4 and 24 h.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7494448 TI - Nicotine elicits changes in expression of adrenal catecholamine biosynthetic enzymes, neuropeptide Y and immediate early genes by injection but not continuous administration. AB - Nicotine, a major pharmacologically active component of tobacco smoke, is generally believed to be one of the factors responsible for the deleterious consequences of cigarette smoking. Nicotine activates the sympathoadrenal system and increases the synthesis and release of catecholamines into circulation. In this study we show that single and repeated injections of nicotine increase the expression of tyrosine hydroxylase (TH), a rate limiting enzyme in the catecholamine biosynthetic pathway. These treatments also regulated the expression of dopamine beta-hydroxylase (DBH) and neuropeptide Y (NPY) in rat adrenals. The effect of nicotine on several transcription factors in the adrenal medulla was examined. Nicotine administration by injection increased the phosphorylation of CREB and induced c-Fos protein without affecting members of the jun family. In contrast to the results with injections, continuous infusion via osmotic pumps did not affect any of these parameters. These data indicate that activation of several transcription factors and increased expression of TH, DBH, and NPY is dependent on the mode of nicotine administration. PMID- 7494450 TI - Molecular analysis of the multiple Golf alpha subunit mRNAs in the rat brain. AB - The alpha subunit of GTP-binding protein Golf (G alpha olf), identified in the olfactory epithelium, in which it is supposed to mediate odorant-generated adenylyl cyclase activations, is much more expressed in the striatum than G alpha s, the classical stimulatory G protein alpha subunit (Herve et al., J Neurosci., 13 (1993) 2237-2248). Four species of G alpha olf messenger RNA (mRNA) were observed by Northern blot in the rat striatum. Analysis of striatal G alpha olf cDNA clones demonstrated that despite extensive variations in their 5' and 3' untranslated regions, these four G alpha olf mRNAs encode the same G alpha olf polypeptide. One of the four mRNA species, which was selectively observed in the striatum, is generated by a novel promoter whose activity was barely detectable in the olfactory epithelium. Surprisingly, this promoter generates an unexpectedly high proportion of transcripts in which the first intron is unspliced. The retention of intron was found to abolish the translation of G alpha olf mRNA in the reticulocyte lysate system, suggesting that it could be involved in the down regulation of G alpha olf expression in the striatum. Furthermore, a new polyadenylation site with a non canonical sequence, AATACA, was found to be responsible of the two shortest forms of G alpha olf mRNA. In conclusion, we have shown that the G alpha olf proteins present in the striatum and the olfactory epithelium are identical and that multiple variations in the untranslated parts of the mRNAs could affect G alpha olf expression. PMID- 7494452 TI - Regional distribution of messenger RNA encoding the insulin-like growth factor type 2 receptor in the rat lower brainstem. AB - Distribution of messenger RNA (mRNA) encoding the insulin-like growth factor (IGF) type 2 receptor (IGF2R) is investigated in the rat lower brainstem by in situ hybridization histochemistry. Cells with IGF2R mRNA are distributed widely in a region-specific manner. It is expressed in: (1) motor nuclei such as the oculomotor nucleus, trochlear nucleus, motor trigeminal nucleus, abducens nucleus, facial nucleus, ambiguus nucleus, dorsal motor nucleus of vagus and hypoglossal nucleus; (2) several sensory-related nuclei like the mesencephalic trigeminal nucleus, ventral nucleus of the lateral lemniscus, lateral and spinal vestibular nuclei, ventral and dorsal cochlear nuclei and nucleus of the trapezoid body; and (3) other regions such as the red nucleus, dorsal raphe nucleus, pontine nuclei, three cerebellar nuclei (medial, interposed and lateral), Purkinje cells, cells in the granular layer of the cerebellum, locus coeruleus, several areas of the reticular nucleus and area postrema. PMID- 7494453 TI - Nicotine regulates nicotinic cholinergic receptors and subunit mRNAs in PC 12 cells through protein kinase A. AB - To understand the up-regulation of neuronal nicotinic cholinergic receptors (nAcChRs) that results from chronic in vivo treatment with nicotine, we studied the effect of nicotine on [3H]nicotine binding sites on PC 12 cells. PC 12 cells were grown in nicotine hemisulfate (10(-6) to 10(-3) M) or vehicle for 7 days, and specific [3H]nicotine binding was measured. Nicotine (10(-6) to 10(-4) M) dose-dependently increased specific binding by up to 2.6-fold over basal levels in 5-7 days, whereas a 10(-3) M concentration failed to do so. In contrast, [3H]nicotine binding to PC 12 cell mutants (A126.1B2 and A123.7), deficient in cAMP-responsive protein kinase A Types I and/or II, was unaffected by nicotine. Northern gel analysis of nAcChR subunit mRNAs from wild type PC 12 cells showed that the mRNA encoding the dominant agonist-binding subunit, alpha 3, was significantly reduced by nicotine, as early as 4 h after treatment, whereas mRNA for the structural beta 2 subunit was slightly increased. In contrast, the alpha 3 subunit mRNA from the PC 12 cell mutant A123.7 was not significantly decreased after 4 h and 7 days of nicotine treatment. These studies indicate that nicotine up-regulates expression of nAcChRs on wild type PC 12 cells and reduces the content of alpha 3 subunit mRNA; these effects require an intact protein kinase A system. The divergent effects of nicotine on the nAcChR compared to its alpha 3 subunit mRNA suggests that enhanced expression of nicotinic receptors may not involve synthesis of new receptor subunit proteins. PMID- 7494451 TI - Regulation of hemicholinium-3 sensitive choline uptake in Xenopus laevis oocytes by the second C2 domain of synaptotagmin. AB - A size-fractionated torpedo electric lobe cDNA library was screened for the neuronal choline transporter by functional expression in oocytes. A clone, TLC2B, was isolated that induced a component of choline uptake that was hemicholinium-3 sensitive and inhibited by the substitution of lithium for sodium at low choline concentrations. However, [3H]choline uptake by both injected and non-injected oocytes were characterized by high affinity constants, suggesting that TLC2B could be affecting a native choline transporter. Indeed, hemicholinium-3 sensitive choline uptake could also be induced by preincubation of non-injected oocytes with a protein kinase C inhibitor, H-7. By sequence analysis and immuno precipitation, the peptide produced by injection of TLC2B cRNA was identified as a soluble 24 kDa C-terminal fragment of the neuronal protein, synaptotagmin. Full length synaptotagmin was, however, ineffective in the functional test. The peptide encoded by TLC2B corresponds to the second protein kinase C-homologous domain of torpedo synaptotagmin, and like other soluble C2 domain peptides, was capable of calcium-dependent translocation to membranes. Its action on choline uptake in oocytes was, however, abolished by the addition of calcium in the presence of a calcium ionophore. These results suggest that the interaction of certain C2 domains, such as the C-terminal domain of synaptotagmin, with more specific targets may be anulled in the presence of calcium due to its absorption to membrane phospholipids. PMID- 7494454 TI - Down-regulation of AMPA-type glutamate receptor gene expression during goldfish optic nerve regeneration. AB - An AMPA-type glutamate receptor cDNA, GFGR52, was cloned from a goldfish retinal cDNA library. GFGR52 is highly homologous to the AMPA D-FLOP-type glutamate receptor. In situ hybridization revealed GFGR52 gene expression in both retinal ganglion cells and optic tectum. This expression was reduced during optic nerve regeneration and this decreased level of gene expression lasted until the reinitiation of synaptogenesis. These findings suggest that interactions between optic nerve and its target, the optic tectum are necessary for maintaining AMPA D type glutamate receptor gene expression in both retinal and tectal neurons. PMID- 7494455 TI - Effect of reserpine treatment on enkephalin mRNA level in the rat striatum: an in situ hybridization study. AB - We investigated the molecular mechanisms responsible for the preproenkephalin A mRNA increase following catecholamine depletion by reserpine using quantitative in situ hybridization at the cellular level. Macroscopic analysis showed that short term reserpine treatment increases the preproenkephalin A mRNA level in the rat striatum to +40.2 +/- 9%. Microautoradiography analysis demonstrated different increases in the preproenkephalin A mRNA level in different parts of the striatum: +124 +/- 22% in the dorso-median striatum, +131 +/- 19% in the dorso-lateral striatum, +119 +/- 8% in the ventro-lateral striatum and +75 +/- 6% in the ventro-median striatum. We found no difference in the number of cells expressing PPA mRNA in reserpine treated rats suggesting that these increases are only due to an increase in the number of mRNA expressed by cell. PMID- 7494456 TI - Dual role of K+ and Na+ on the transport of [3H]-gamma-aminobutyric acid by synaptic plasma membrane vesicles. AB - The influence of the monovalent cations (Na+ and K+) and of the electrical gradient on the high-affinity [3H]-gamma-aminobutyric acid ([3H]GABA) transport was investigated in synaptic plasma membrane (SPM) vesicles isolated from sheep brain cortex. This process specifically requires internal K+, since when it is replaced by Li+, the delta psi remains of the same order of magnitude, but no uptake of [3H]GABA occurs. The influence of the external Na+ concentration on the rate of [3H]GABA uptake suggests that this mechanism exhibits two components, whose characteristics are determined by the delta psi. Depolarization reduces the Jmax of [3H]GABA influx and enhances the binding of Na+ associated to [3H]GABA transport. Nevertheless, depolarization does not affect the K0.5 of binding sites for Na+ and the stoichiometry of translocation. These results suggest that intravesicular K+ and external Na+ have a dual role on the mechanism of [3H]GABA uptake: K+ acts directly on the carrier and determines the membrane polarization; Na+ is cotransported with GABA and, according to the polarization state of the membrane, it modulates the operation of the carrier in its inward GABA translocation. PMID- 7494458 TI - Molecular cloning of a novel protein regulated by opioid treatment of NG108-15 cells. AB - A new cDNA clone, NGD5, has been identified from a subtraction cDNA library constructed with mRNA isolated from control neuroblastoma x glioma NG108-15 cells and cells treated for 48 h with the delta-opioid agonist, D-Ala2, D-Leu5 enkephalin (DADLE). NGD5 mRNA is decreased, in a naloxone-reversible manner, upon long-term treatment of NG108-15 cells with DADLE, indicating that this clone may be related to opioid receptor function. Northern analysis indicates that NGD5 mRNA is expressed in rat brain. Two similar nearly full-length NGD5 clones, NGD5A and NGD5B, were isolated from a lambda gt10 NG108-15 cDNA library and sequenced. The predicted 40-kDa peptide encoded for by the NGD5 cDNA has no significant homology to the recently cloned mu, delta or kappa opioid receptors nor to any other known proteins. PMID- 7494459 TI - Effects of chronic social stress on tyrosine hydroxylase mRNA and protein levels. AB - Males housed in mixed sex groups quickly form dominance hierarchies; subordinates can be further subdivided into stress responsive subordinates (SRS) and non responsive subordinates (NRS) based on corticosterone responses to a novel stressor. Tyrosine hydroxylase (TH) mRNA levels measured with in situ hybridization were elevated in locus coeruleus (LC) of NRS compared to singly or pair-housed controls; NRS also had higher TH levels than dominants. TH protein levels determined by immunoautoradiography were also higher in LC of NRS and SRS versus pair-housed controls. PMID- 7494457 TI - Opioid mu- and kappa-receptor mediate phospholipase C activation through Gi1 in Xenopus oocytes. AB - In the Xenopus oocytes expressing mu- or kappa-opioid receptors, agonist-induced currents were observed only when the oocyte was coinjected with Gi1 alpha RNA and pretreated with K-252a, a potent inhibitor of protein kinases. The evoked currents were abolished by intracellular injection of EGTA or inositol 1,4,5 trisphosphate and the current-voltage relationship revealed that they are mediated through typical calcium-dependent chloride channels. These findings suggest that the mu- and kappa-receptors mediate phospholipase C activation through Gi1 alpha, and that these receptor mechanisms including downstream signalings might be inhibited by phosphorylation in vivo in the Xenopus oocyte. PMID- 7494460 TI - Proteolysis of spectrin by calpain accompanies theta-burst stimulation in cultured hippocampal slices. AB - Tests were carried out to determine if repetitive bursts of afferent stimulation activate calpain, a calcium-dependent protease hypothesized to be involved in the production of long-term potentiation. Antibodies against a stable breakdown product that results from proteolysis of spectrin by calpain were used to identify sites of enzyme activation in cultured hippocampal slices. Slices in which theta-burst stimulation was applied to the Schaffer collateral fibers had pronounced accumulations of breakdown product that were restricted to field CA1, the zone innervated by the stimulated axons. Labelling occurred in the form of scattered puncta and was also present in dendritic processes. The extent of these effects was correlated (r = 0.73) with the amount of theta-burst stimulation delivered. Control slices or those receiving low frequency stimulation had variable, but uniformly lower, amounts of breakdown product and were clearly distinguishable from those given theta bursts. Statistical analyses using a six point rating scheme confirmed this point (P < 0.001). These results satisfy an essential prediction of the hypothesis that calpain plays an important role in the induction of long-term potentiation. PMID- 7494462 TI - The sap47 gene of Drosophila melanogaster codes for a novel conserved neuronal protein associated with synaptic terminals. AB - Proteins expressed specifically in neurons and transported to synaptic terminals are likely to constitute important molecular elements of nervous system function. In an effort to characterize synapse-associated proteins (SAPs) of Drosophila, we have isolated from a hybridoma library several monoclonal antibodies (MABs) that selectively stain synaptic terminals in immunohistochemical preparations. MAB nc46 binds to most but not all synaptic terminals of the Drosophila nervous system, it also recognizes a protein with homologous distribution in other dipteran flies and binds to large parts of fish CNS. In Western blots the antibody labels a Drosophila brain protein of 47 kDa and cross-reacts with brain proteins from several species including insects, fish, mouse and man. From these data we conclude that the corresponding gene has been conserved in evolution at least among diptera. Using MAB nc46 and expression cloning we have identified the 'sap47' gene coding for the 'synapse-associated protein of 47 kDa' of Drosophila melanogaster. Sequence analysis of genomic and cDNA clones reveals the intron exon structure of the gene and characterizes the complete open reading frames of two alternatively spliced transcripts. The sap47 gene is located in 89A8-B3 on chromosome 3R and codes for two almost identical inferred polypeptides of 347 and 351 amino acids with no significant sequence homology to known proteins. PMID- 7494461 TI - Selective localization of amyloid precursor-like protein 1 in the cerebral cortex postsynaptic density. AB - Senile plaques, a hallmark of Alzheimer's disease (AD), contain amyloid beta peptide (A beta), which is generated from the larger amyloid beta protein precursor (APP). In addition to APP, several APP-related proteins have been recently identified in different organisms, including Drosophila amyloid precursor protein-like protein (APPL). Deficiency of APPL causes behavioral deficits in Drosophila, implicating a role in brain function. Moreover, mouse and human cDNA clones encoding amyloid precursor-like proteins (APLP1 and APLP2) have been identified and exhibit extensive sequence similarity to the APPL and APP genes. To define the potential role of APLP in the mammalian brain, we sought to directly localize APLP1 within the complex cortical synaptic structure. We focused on the postsynaptic density (PSD), which appears to be central to synaptic function. We now report that the 90 kDa APLP1, the first known APLP, is localized to the PSD from rat and human cerebral cortex. APLP1 increased during cortical synaptic development, suggesting a role in synaptogenesis or synaptic maturation. In contrast, APP was predominantly expressed in the synaptic membrane fraction, but was barely detectable in the PSD, including different subcellular distributions of APP and APLP1. Our observations raise the possibility that APLP1, a homologue of APPL, which appears to be necessary for normal behavior in Drosophila, participates in brain synaptic function in mammals. PMID- 7494463 TI - Induction of VGF mRNA in neurons of the rat nucleus tractus solitarius and the dorsal motor nucleus of vagus in duodenal ulceration by cysteamine. AB - To investigate the possible role of the brainstem in cysteamine-induced peptic ulceration, we examined the expression of VGF mRNA, which is induced in PC12 cells following application of nerve growth factor [23], in the nucleus tractus solitarius (NTS)/dorsal motor nucleus of vagus (DMV) complex of the medulla oblongata by in situ hybridization histochemistry. In control saline-treated rats, weak VGF mRNA signals were only rarely detected in neurons of the NTS and none were observed in those in the DMV. After 12 h of cysteamine administration (450 mg/kg, s.c.), the time at which duodenal ulcer was detected in all cases, heavily labeled VGF mRNA-expressing neurons appeared in the NTS and DMV. By quantitative analysis on macroautoradiogram, the VGF mRNA signals of the NTS/DMV complex in cysteamine-treated rats were twice as much as those in saline-treated rats. In situ hybridization histochemistry combined with the use of the retrograde neuronal tracer cholera toxin-B subunit revealed that the induced VGF mRNA-expressing neurons of the DMV projected directly to the stomach. The present results suggest that ulceration accompanies the induction of VGF mRNA in neurons of vagal afferent and efferent areas of the brainstem. PMID- 7494464 TI - Expression of platelet-derived growth factor B-chain in the mature rat brain and pituitary gland. AB - For better understanding of the role of platelet-derived growth factor (PDGF) B chain in the brain, the expression of PDGF B-chain was studied in the mature rat brain at both protein and mRNA levels, by assay of PDGF B-chain-related mitogenic activity, Northern blot, in situ hybridization and immunohistochemistry. It was shown that (1) mature rat brain contained substantial PDGF B-chain-related mitogenic activity, (2) significant amounts of two sizes of transcripts (3.5 kb, widely, and 2.6 kb, weakly and in narrower areas) were expressed in the brain, and (3) the transcripts were localized in ubiquitous neurons by in situ hybridization, with the strongest signal in hippocampal pyramidal cells, which distribution almost corresponded with that of the immunoreactive products. The abundant neuronal localization of the transcript and protein, as well as the neuronal expression of the receptor reported elsewhere, suggests the role of the growth factor in neuronal cells as a neuronal regulatory and/or trophic agent acting by autocrine loop or by neuron to neuron interaction. However, there was an apparent discrepancy in part, in the distribution between transcripts and immunoreactivity; that is, transcripts were expressed intensely in the intermediate pituitary lobe with only a scattered immunoreactivity, and the opposite situation was observed in the accessory olfactory nerve and posterior pituitary lobe. This might suggest that PDGF B-chain is transported or secreted in these foci. PMID- 7494465 TI - Cortisol up-regulates corticotropin releasing factor gene expression in the fetal ovine brainstem at 0.70 gestation. AB - Glucocorticoids are important for the development of the central nervous system. In the ovine fetus, increased levels of plasma cortisol at term provide a stimulus to initiate parturition. CRF is central to this event in that it is one of the main modulators of the hypothalamic-pituitary-adrenal (HPA) axis. The purpose of the present study was to determine the effect of physiological increases in fetal plasma cortisol levels on corticotropin-releasing factor (CRF) gene expression in the developing ovine brain. Fetal plasma cortisol levels were chronically elevated at 0.70 gestation (100 days) to physiological levels found at 0.90 gestation (130 days; term 145 +/- 2 days) when glucocorticoid-induced maturational changes are known to occur in the HPA axis. The 3' end of the ovine CRF gene encodes 4 putative polyadenylation (poly(A)) signals that may post transcriptionally regulate gene expression through stability, translation and localization of the mRNA in a temporal and spatial manner. To determine whether CRF mRNA levels or poly(A) site usage are differentially regulated by cortisol in a region-specific manner, we used an RNase protection assay with an antisense CRF RNA probe from the 3' coding and untranslated regions of the gene to quantify changes in mRNA levels in the hypothalamus (Hypo), hippocampal-amygdala complex (H and A), frontal cerebral cortex (FCC) and brainstem. Our novel finding was a 3.5-fold increase in CRF mRNA levels in the medulla oblongata of fetuses from the cortisol group compared to those from the saline group (P = 0.001). CRF mRNA levels in the Hypo, H and A and FCC did not change significantly in fetuses from the cortisol group.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7494466 TI - Quantitative measurement of neuronal calbindin-D28k by radioimmunocytochemistry. AB - The calcium-binding protein calbindin-D28k (CaBP) has been localized in high concentration in several neuronal populations within the CNS and is believed to act as an intracellular calcium buffer. There has been much interest and speculation concerning its potential neuroprotective function. A radioimmunocytochemistry (RIC) technique for the cellular quantitation of protein has been applied to quantitative measurement of neuronal CaBP in vivo and in vitro. The method permits cellular comparison of CaBP content within tissue sections or cells in culture. Through the use of specific primary antibody, 35S labeled secondary antibody, and photographic emulsion, RIC combines the simplicity of standard immunocytochemical procedures with the sophistication and power of in situ hybridization, autoradiography, and image analysis. CaBP levels are expressed as mean +/- S.E.M. silver grains/cell. CaBP content has been measured and compared in mouse cerebellar Purkinje cells (56.5 +/- 6.9 grains/cell), granule cells of the hippocampal dentate gyrus (10.3 +/- 2.1 grains/cell), midline ventral tegmental neurons (11.6 +/- 2.9 grains/cell), and human SH-SY-5Y neuroblastoma cells in culture (5.1 +/- 0.9 grains/cell). As measured by RIC, mouse cerebellar Purkinje cells contain approximately 5-fold more CaBP than granule cells of the hippocampal dentate gyrus/midline ventral tegmental neurons and 10-fold more CaBP than cultured human SH-SY-5Y neuroblastoma cells. Assay reproducibility was demonstrated by comparison of adjacent sections which yielded a 3-9% intra-assay variability. Results were validated and confirmed by comparison to previous radioimmunoassay studies which indicated similar ratios of CaBP levels between brain regions/cell types.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7494467 TI - Identification of two alternatively spliced transcripts of STEP: a subfamily of brain-enriched protein tyrosine phosphatases. AB - A brain-enriched protein tyrosine phosphatase termed STEP46 (striatal enriched phosphatase) was previously isolated and characterized. Immunological studies with a STEP monoclonal antibody recognized several STEP-immunoreactive proteins, and suggested that additional STEP-related polypeptides existed. This study reports the isolation of two alternatively spliced transcripts of the STEP gene. One of these, STEP20 (with a predicted molecular mass of 20 kDa) was further characterized and found to lack the conserved tyrosine phosphatase domain. Northern analysis detected a 2.8 kb STEP20 message in mouse brain. The second alternatively spliced transcript, STEP61, has a 5'-extended open reading frame that encodes a protein with a predicted molecular mass of 61 kDa and contains a single tyrosine phosphatase domain. The exon-intron organization responsible for the novel STEP20 and STEP61 sequences was determined in the mouse STEP genomic DNA. We propose that the original STEP46, along with STEP20 and STEP61, are members of a brain-enriched subfamily of protein tyrosine phosphatases, and that STEP isoforms may have distinct functions within the central nervous system. PMID- 7494468 TI - The distribution of splice variants of the NMDAR1 subunit mRNA in adult rat brain. AB - Regional variation in the alternative splice forms of the NMDAR1 subunit mRNA was investigated by in situ hybridization in the adult rat brain, using radiolabelled splice-specific oligonucleotide probes. Each splice variant was detected in an individual distribution. The NMDAR1-a and NMDAR1-2 forms were widely and abundantly distributed throughout the brain, except for the inferior colliculus. The NMDAR1-b and NMDAR1-4 variants were located in similar patterns in fewer areas (e.g. parietal cortex, hippocampus CA3, thalamus, inferior colliculus, cerebellar granule cells). In contrast, the NMDAR1-1 forms were distributed in a pattern approximately complementary in the forebrain to that of NMDAR1-4 (weakly expressed in thalamus and inferior colliculus). The NMDAR1-3 variants were not abundant in any structure. Considerable overlap of the in situ hybridization images was noted, so all eight splice combinations are possible in heterogenous distributions. Correlation of the distribution of NMDAR1 mRNA splice forms with functional analyses of heteromeric recombinant receptors will be necessary to ascertain if alternative splicing of the NMDAR1 subunit can account for some of the known heterogeneity of endogenous NMDA receptors. PMID- 7494469 TI - Programmed cell death in bacteria: proteic plasmid stabilization systems. AB - Bacterial plasmids are stabilized by a number of different mechanisms. Here we describe the molecular aspects of a group of plasmid-encoded gene systems called the proteic killer gene systems. These systems mediate plasmid maintenance by selectively killing plasmid-free cells (post-segregational killing or plasmid addiction). The group includes ccd of F, parD/pem of R1/R100, parDE of RP4/RK2, and phd/doc of P1. All of these systems encode a stable toxin and an unstable antidote. The antidotes prevent the lethal action of their cognate toxins by forming tight complexes with them. The antidotes are degraded by cellular proteases. Thus, the different decay rates of the toxins and antidotes seem to be the molecular basis of toxin activation in plasmid-free cells. The operons encoding the toxins and antidotes are autoregulated at the level of transcription either by a complex formed by the toxins and the cognate antidotes or by the antidote alone. The cellular targets of the killer proteins have been determined to be DNA gyrase in the case of ccd of F and DnaB in the case of parD of R1. Surprisingly, the Escherichia coli chromosome encodes at least two of these peculiar gene systems. PMID- 7494471 TI - Deletion of DNA lying close to the glkA locus induces ectopic sporulation in Streptomyces coelicolor A3(2). AB - Streptomyces coelicolor A3(2) J1668 sporulated ectopically in the substrate hyphae (the Esp phenotype) with the same time course as sporulation in the aerial hyphae. Examination of related strains implied that the Esp phenotype was caused by the deletion of DNA that lies close to, but is distinct from, the glucose kinase gene (glkA). Co-transduction of the Esp phenotype with the deletion present in J1668 confirmed this hypothesis. The size of the deletion was found to be 7.4 kb. Construction of a strain carrying both the J1668 deletion and a whiG mutation demonstrated that the Esp phenotype depends on at least one of the genes required for the differentiation of aerial hyphae into spores. PMID- 7494470 TI - Comparison of ccd of F, parDE of RP4, and parD of R1 using a novel conditional replication control system of plasmid R1. AB - A number of plasmid-encoded gene systems are thought to stabilize plasmids by killing plasmid-free cells (also termed post-segregational killing or plasmid addiction). Here we analyse the mechanisms of plasmid stabilization by ccd of F, parDE of RP4 and parD of R1, and compare them to hok/sok of R1. To induce synchronous plasmid loss we constructed a novel plasmid replication-arrest system, which possesses the advantage that plasmid replication can be completely arrested by the addition of IPTG, a non-metabolizable inducer. Using isogenic plasmid constructions we have found, for the first time, consistent correlation between the effect on steady-state loss rates and the effect on cell proliferation in the plasmid replication-arrest assay for all three systems. The parDE system had the most pronounced effect both on plasmid stabilization and on plasmid retention after replication arrest. In contrast, ccd and parD both exhibited weaker effects than anticipated from previously published results. Thus, our results indicate that the function and efficiencies of some of the systems should be reconsidered. Our results are consistent with the previously postulated hypothesis that ccd and parDE act by killing plasmid-free segregants, whereas parD seems to act by inhibiting cell division of plasmid-free segregants. PMID- 7494472 TI - Characterization of a new rho mutation that relieves polarity of Mu insertions. AB - We report the identification and characterization of a new rho mutation, rho614, that relieves polarity of Mu insertions in Escherichia coli. The mutation was identified by its ability to suppress the polarity of the Mu-mediated phi(lamB' 'lacZ)hyb61-4 fusion that is located at codon four of the lamB signal sequence. The rho614 mutation alters residue 80 in the proposed RNA-binding domain of Rho and is recessive to wild-type rho. We suggest that in the presence of the rho614 allele transcripts initiated at the Mu promoter PcM fail to terminate at presumptive Rho-dependent termination sites, namely rut1 and rut2, and continue into the 3' 'lamB gene allowing a LamB+ phenotype. This contention is supported by deletion analysis of the region and the observation that insertional inactivation of genes that reduce transcription from PcM, clpP (ATP protease), himA (IHF-alpha), and himD (IHF-beta), block the LamB+ phenotype. rho614, rho4 and rho201 alleles suppress the polarity of a malK::Mu insertion on the downstream lamB gene. However, the polarity of the phi(lamB'-'lacZ)hyb61-4 insertion is only suppressed by the rho614 mutation. We propose that the rho614 mutation allows suppression of transcriptional polarity without interfering with translation initiation signals of the truncated 'lamB gene. In addition to identifying a new rho mutation and Rho-dependent terminator sequence, this system provides a means of studying Rho protein/terminator relationships through the identification of new classes of rho mutations. PMID- 7494473 TI - Identification of a novel virulence gene, virA, on the large plasmid of Shigella, involved in invasion and intercellular spreading. AB - A novel virulence gene (virA) was identified upstream of the virG gene on the large plasmid of Shigella flexneri 2a YSH6000. Characterization of virA mutants infecting MK2 epithelial cell monolayers revealed that their invasive capacity was decreased to less than one fifth of the wild-type level. Nevertheless, the bacteria were capable of expressing and secreting IpaB, IpaC and IpaD proteins. The virA mutants were also impaired in their ability to spread intercellularly, since the bacteria gave rise to a small number of foci in a focus-plaque-forming test with MK2 cells. Although virG expression was slightly decreased in the virA mutants, introduction of a cloned virG gene into a virA mutant, N1945, failed to restore spreading ability. Although, introduction of a cloned virA gene into N1945 restored invasiveness and spreading ability, the reduced virG transcription level was not affected, indicating that the reduced virG expression in virA mutants does not play a major role in defective intercellular spreading. The nucleotide sequence of the virA region revealed that the virA gene was located 528 bp upstream of the virG gene, in the opposite orientation. The deduced amino acid sequence of the VirA protein indicated a 44.7 kDa protein with no homology to known proteins. The VirA protein was secreted into the culture supernatant, a process that required the Mxi and Spa loci. The expression of virA was under the control of the virB gene, the positive regulator of the ipa, mxi and spa operons. These results indicate that virA is a new member of the invasion regulon directed by virB and that the VirA function is involved in invasion and intercellular spreading. PMID- 7494474 TI - Glucose lowers CRP* levels resulting in repression of the lac operon in cells lacking cAMP. AB - CRP-cAMP-dependent operons of Escherichia coli can be expressed in cells lacking functional adenylate cyclase when they carry a second-site mutation in the crp gene (crp*). It is known that the expression of these operons is repressed by glucose, but the molecular mechanism underlying this cAMP-independent catabolite repression has been a long-standing mystery. Here we address the question of how glucose inhibits the expression of beta-galactosidase in the absence of cAMP. We have isolated several mutations in the crp gene that confer a CRP* phenotype. The expression of beta-galactosidase is reduced by glucose in cells carrying these mutations. Using Western blotting and/or SDS-PAGE analysis, we demonstrate that glucose lowers the cellular concentration of CRP* through a reduction in crp* mRNA levels. The level of CRP* protein correlates with beta-galactosidase activity. When the crp promoter is replaced with the bla promoter, the inhibitory effect of glucose on crp* expression is virtually abolished. These data strongly suggest that the lowered level of CRP* caused by glucose mediates catabolite repression in cya- crp* cells and that the autoregulatory circuit of the crp gene is involved in the down-regulation of CRP* expression by glucose. PMID- 7494475 TI - An Agrobacterium virulence factor encoded by a Ti plasmid gene or a chromosomal gene is required for T-DNA transfer into plants. AB - Mutagenesis of the vir region on the Ti plasmid of Agrobacterium tumefaciens revealed a new locus, virJ, that is induced by the plant-wound signal molecule, acetosyringone (AS). virJ lies between virA and virB, and is transcribed in the same direction. The amino acid sequence of virJ is similar to a region of a previously characterized chromosomal gene, acvB, required for virulence. virJ can complement the avirulent phenotype of an acvB mutant, indicating that virJ and acvB encode the same factor required for tumorigenesis. Southern analysis revealed that virJ is present on the Ti plasmid of an octopine but not a nopaline strain whereas acvB is present on the chromosomes of both octopine and nopaline strains. While virJ is regulated by AS under the control of the virA/virG two component regulatory system, acvB is not induced by AS. VirJ possesses a putative signal peptide and was found predominantly in the periplasmic fraction. The strain lacking both acvB and virJ had an impaired ability to transfer T-DNA into plant cells, suggesting that the factor encoded by virJ or acvB is required for T DNA transfer from A. tumefaciens to plant cells. acvB is the first chromosomal gene implicated in T-DNA transfer, but whether it functions specifically for this process is not clear. We hypothesize that virJ evolved from acvB, presumably for a more specialized role in tumorigenesis. PMID- 7494477 TI - Evidence that the transcriptional activator Spo0A interacts with two sigma factors in Bacillus subtilis. AB - The transcriptional regulator Spo0A activates transcription from two types of promoters. One type of promoter is used by RNA polymerase containing sigma A, whereas the other type is used by RNA polymerase containing sigma H. There are Spo0A-binding sites near the -35 region of both types of promoters. It has been reported that some transcriptional regulators that bind near the -35 regions of promoters directly interact with the sigma subunit of RNA polymerase. Therefore, we looked for evidence that Spo0A interacts with both sigma factors by searching for single amino acid substitutions in these factors that specifically prevent expression from Spo0A-dependent promoters, but that do not decrease activity of Spo0A-independent promoters. Two such amino acid substitutions were isolated in sigma A and one was isolated in sigma H. The amino acid substitutions in sigma A prevented expression from the Spo0A-activated promoters, spoIIG and spoIIE, but expression was not impaired from the Spo0A-independent, sigma A-dependent promoter tms or from the Spo0A-activated, sigma H-dependent promoter, spoIIA. The amino acid substitution in sigma H prevented expression from the spoIIA promoter but not from the Spo0A-independent promoter, citGp2, which is used by sigma H-RNA polymerase. All of these amino acid substitutions occur in the carboxyl terminus of the sigma factors. These amino acid substitutions may define the sites of contact between the sigma factors and Spo0A. The ability of response regulators such as Spo0A to interact with multiple sigma factors may increase the variety of responses made by bacteria using a limited number of transcription factors. PMID- 7494476 TI - The -16 region, a vital sequence for the utilization of a promoter in Bacillus subtilis and Escherichia coli. AB - The promoter (amyP) of the Bacillus subtilis alpha-amylase gene, which is recognized by E sigma A, has a three out of six match to the consensus promoter in both the -35 and -10 hexamers. Oligonucleotide-directed mutagenesis was used to identify important bases for promoter utilization in the spacer sequence between the hexamers. Mutations in the sequence TGTG extending from positions -18 to -15 (the -16 region) caused a 5-94-fold decrease in alpha-amylase production. A G-C transversion at position -15 was the most detrimental mutation: it essentially eliminated amyP utilization in B. subtilis and in Escherichia coli. Mutating the -35 and -10 hexamers to the E sigma A consensus promoter increased alpha-amylase production 56-fold in B. subtilis and fivefold in E. coli. Introducing the -15 G to C transversion into the consensus promoter reduced alpha amylase production threefold, in contrast to the 94-fold reduction for the wild type promoter in B. subtilis. The -15 G to C transversion did not reduce alpha amylase synthesis directed by the consensus promoter in E. coli. The alpha amylase gene is subject to two forms of transcriptional regulation: catabolite repression and temporal regulation. None of the mutants constructed in this study had any effect on either type of regulation. The -16 region, especially the G at position -15, appears to be moderately conserved in B. subtilis and in other Gram positive organisms and weakly conserved in E. coli. The evidence suggests that the -16 region is an additional region of E sigma A promoters in B. subtilis and E sigma 70 promoters in E. coli, essential in some weak promoters such as the alpha-amylase promoter but, of little benefit to very strong promoters. PMID- 7494478 TI - A transcription terminator signal necessary for plasmid ColIb-P9 replication. AB - Replication of the IncI alpha plasmid ColIb-P9 requires the repZ gene, which encodes an essential, unstable initiator protein termed RepZ. Although many functional features of the ColIb-P9 replicon resemble those of structurally unrelated IncFII plasmids R1 and NR1, the role of transcription of repZ towards the replication origin is poorly understood. Using a series of deletion and substitution mutants of the ColIb-P9 replicon, we found that RepZ prefers to act in cis and that a spacer sequence between repZ and the origin is required for replication. This spacer element, referred to as CIS, retained strong transcription terminator activity. Efficient transcription terminators, whether Rho-dependent or -independent, were capable of replacing CIS function for in vivo replication; ColIb-P9 replicated better as transcription terminated more efficiently within CIS. When the CIS element was substituted for by a strong Rho dependent terminator, such as lambda tR1 or E. coli trp t', in vivo replication of these recombinant replicons became dependent on the Rho factor, in contrast to the authentic ColIb-P9 replicon. PMID- 7494479 TI - Leucine-responsive regulatory protein plays dual roles as both an activator and a repressor of the Escherichia coli pap fimbrial operon. AB - The expression of the pap pilus operon of Escherichia coli is under a phase variation control mechanism in which cells undergo a reversible transition between transcriptionally active (phase ON) and inactive (phase OFF) states. In this study, we explore the roles of leucine-responsive regulatory protein (Lrp) and the histone-like protein H-NS in the regulation of pap phase variation. Our data indicate that the phase OFF state results from repression of the intrinsically active papBA promoter by Lrp and H-NS, each of which can act independently as transcriptional repressors. Lrp requires pap DNA sequences upstream of the papBA promoter for its repressor activity whereas H-NS does not. In contrast, in the ON state, Lrp, in conjunction with PapI, activates pap transcription. This activation is not merely a result of alleviating the H-NS mediated repression, but induces a level of transcription that is eightfold higher than the basal level of transcription from the papBA promoter measured in the absence of both H-NS and Lrp. Analysis of Lrp activation mutants indicates that binding of Lrp to pap DNA sequences is not sufficient for transcription activation, consistent with a model in which an additional domain of Lrp interacts with the transcriptional apparatus. Together, our results show that Lrp functions as a transcriptional activator in phase-ON cells and as a repressor of basal transcription in phase-OFF cells. Because pap phase variation occurs in the absence of H-NS, it is not clear what role this regulatory protein plays in pap gene regulation. PMID- 7494480 TI - Closing in on the toxic domain through analysis of a variant Clostridium difficile cytotoxin B. AB - Strain 1470 is the standard typing strain for serogroup F of Clostridium difficile containing both toxin genes, toxA-1470 and toxB-1470. A polymerase chain reaction (PCR)-based approach to the sequencing of the total toxB-1470 gene identified an open reading frame (ORF) of 7104 nucleotides. In comparison with the previously sequenced toxB of C. difficile VP10463, the toxB-1470 gene has 16 additional nucleotides, 13 within the 5'-untranslated region and three within the coding region. The M(r) of ToxB-1470 is 269,262, with an isoelectric point (IP) of 4.16. The equivalent values for ToxB are M(r) 269,709 and IP 4.13. In comparison with ToxB, ToxB-1470 differs primarily in the N-terminal region between positions 1 and 868 where 148 amino acids residues are changed. The C terminal region between residues 869-2367 is highly conserved with only six amino acid alterations. Dot matrix comparison of ToxB-1470 with ToxA and ToxB reveals the highest homology between ToxB-1470 and ToxB. Thus ToxB-1470 did not originate from recombination between ToxA and ToxB. On cultured endothelial cells, from porcine pulmonary artery, purified ToxB-1470 is less potent than ToxB. The cytopathic effects of ToxB-1470 are indistinguishable from those caused by the lethal toxin (LT) of Clostridium sordellii, but are clearly different from the patterns observed after exposure of endothelial cells to ToxA and ToxB of C. difficile (VPI10463) or alpha-toxin (Tcn alpha) of Clostridium novyi. The LT-like action of ToxB-1470 was not due to altered internalization processes, as microinjection and addition to the medium induced identical effects on the cells.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7494481 TI - Gradient of genomic diversity in the Pseudomonas aeruginosa chromosome. AB - In 545 Pseudomonas aeruginosa strains, mainly collected from patients with cystic fibrosis, SpeI-DraI macrorestriction fragment length diversity was scanned for using probes of known map position on the P. aeruginosa PAO chromosome. Southern analysis of the 60 unrelated clones uncovered a gradient of macrorestriction fragment length polymorphisms (RFLPs) from the origin of replication towards the auxotroph-poor region of the P. aeruginosa genome. Linkage disequilibrium between macrorestriction sites was conserved in the P. aeruginosa population in the region encompassed by the rrn operons. The oriC-reactive SpeI fragment was conserved in nearly all isolates examined. Few fragment length classes were seen for the alg60-, algR- and toxA-encoding SpeI fragments. Fragment size varied within one class by up to 20 kb. Two probes from the auxotroph-poor region detected a broad size range for the SpeI fragment, suggesting extensive genomic diversity in these regions. Subclonal variation of fragment size was detected at all investigated loci in at least one of the analysed clones, but within one particular clone, SpeI-RFLPs were found at only few loci. PMID- 7494482 TI - Multiple homologues of LuxR and LuxI control expression of virulence determinants and secondary metabolites through quorum sensing in Pseudomonas aeruginosa PAO1. AB - In Pseudomonas aeruginosa PAO1, expression of elastase is dependent upon an interaction between the positive transcriptional activator LasR and the auto inducer molecule N-(3-oxododecanoyl)-L-homoserine lactone (OdDHL), the synthesis of which is directed by LasI. Previously we have shown that in PAN067, an elastase-negative mutant of PAO1, elastase production can be restored to some extent by addition of exogenous N-(3-oxohexanoyl)-L-homoserine lactone (OHHL). Here we report that PAN067 is also defective in the production of alkaline protease, haemolysin, cyanide, pyocyanin and autoinducer(s). As neither addition of exogenous OdDHL nor introduction of lasR restored PAN067 to the parental phenotype, we sought to complement PAN067 with PAO1 DNA. From a cosmid library, a 2 kb DNA fragment was identified which re-established production of autoinducer(s) and exoproducts in PAN067. From the nucleotide sequence of this fragment, two genes termed rhIR and rhII were identified. RhII is responsible for autoinducer synthesis and shares 31% homology with LasI; RhIR has been previously identified in P. aeruginosa strain DSM2659 as a regulator of rhamnolipid biosynthesis and shares 28% identity with LasR. These data provide clear evidence that multiple families of quorum-sensing modulons interactively regulate gene expression in P. aeruginosa. PMID- 7494483 TI - Characterisation of the yenI/yenR locus from Yersinia enterocolitica mediating the synthesis of two N-acylhomoserine lactone signal molecules. AB - Yersinia enterocolitica produces compounds capable of transcriptionally activating the Photobacterium fischeri bioluminescence (lux) operon. Using high performance liquid chromatography, high resolution tandem mass spectrometry in conjunction with chemical synthesis, two signal molecules were identified and shown to be N-hexanoyl-L-homoserine lactone (HHL) and N-(3-oxohexanoyl)-L homoserine lactone (OHHL). A gene (yenI) was isolated from Y. enterocolitica and demonstrated to direct the synthesis of both HHL and OHHL. DNA sequence analysis revealed an open reading frame (ORF) of 642 bp encoding a protein (YenI) of 24.6 kDa with approximately 20% identity to the LuxI family of proteins. Northern blot analysis of yenI expression indicated yenI is transcribed as a single gene and 5' transcript mapping of yenI identified a transcriptional start site 89 bp upstream of the ORF. DNA sequence analysis of the region downstream of yenI located a second ORF, termed yenR, with significant homology to the LuxR family of transcriptional activators. An insertion mutation of yenI abolishes HHL and OHHL production, indicating its central role in N-acylhomoserine lactone synthesis in Y. enterocolitica. Transcriptional analysis using a chromosomal yenI::luxAB fusion has demonstrated that yenI is not subject to autoinduction but is expressed constitutively. Whilst production of the Yop proteins in the wild type and in yenI mutants is indistinguishable, two-dimensional SDS-PAGE analysis of total cell proteins indicated that a number of proteins lack the yenI mutant. PMID- 7494484 TI - Identification of a chemotaxis operon with two cheY genes in Rhodobacter sphaeroides. AB - A large chemotaxis operon was identified in Rhodobacter sphaeroides WS8-N using a probe based on the 3' terminal portion of the Rhizobium meliloti cheA gene. Two genes homologous to the enteric cheY were identified in an operon also containing cheA, cheW, and cheR homologues. The deduced protein sequences of che gene products were aligned with those from Escherichia coli and shown to be highly conserved. A mutant with an interrupted copy of cheA showed normal patterns of swimming, unlike the equivalent mutants in E. coli which are smooth swimming. Tethered cheA mutant cells showed normal responses to changes in organic acids, but increased, inverted responses to sugars. The unusual behaviour of the cheA mutant and the identification of two homologues of cheY suggests that R. sphaeroides has at least two pathways controlling motor activity. To identify functional similarity between the newly identified R. sphaeroides Che pathway and the methyl-accepting chemotaxis protein (MCP)-dependent pathway in enteric bacteria, the R. sphaeroides cheW gene was expressed in a cheW mutant strain of E. coli and found to complement, causing a partial return to a swarming phenotype. In addition, expression of the R. sphaeroides gene in wild-type E. coli resulted in the same increased tumbling and reduced swarming as seen when the native gene is overexpressed in E. coli. The identification of che homologues in R. sphaeroides and complementation by cheW suggests the presence of MCPs in an organism previously considered to use only MCP-independent sensing. The MCP dependent pathway, appears conserved.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7494485 TI - A cellulase/xylanase-negative mutant of Streptomyces lividans 1326 defective in cellobiose and xylobiose uptake is mutated in a gene encoding a protein homologous to ATP-binding proteins. AB - The uptake of monosaccharides (glucose and xylose) and disaccharides (cellobiose and xylobiose) was evaluated in the Streptomyces lividans mutant strain 10-164. The pleiotropic mutation had no effect on glucose uptake; however, the Vmax of xylose uptake was decreased 10-fold as compared to the wild-type strain, S. lividans 1326, and the transport system of cellobiose and xylobiose, the putative inducers of the cellulase and xylanase genes, was completely abolished resulting in a cellulase/xylanase-negative mutant. An accumulation of xylose and glucose in culture media was observed when the mutant was grown on xylobiose and cellobiose, respectively. Cell-associated beta-glucosidase and low levels of extracellular beta-glucosidase were detected in both strains. When gluconolactone, a beta glucosidase inhibitor, was added to the medium there was no uptake of cellobiose or release of glucose by the mutant strain, whereas the uptake of cellobiose by the wild-type strain was not significantly affected. It is thus proposed that the active transport system for cellobiose and xylobiose is affected in mutant strain 10-164. Glucose and xylose production from disaccharide hydrolysis are due to beta-glucosidase and beta-xylosidase activities, which sustain the growth of the mutant strain. Clones complementing the mutation were isolated from a gene bank constructed using mutant strain 10-164. The msiK gene codes for MsiK, a 40 kDa multiple sugar import protein, which belongs to the family of ATP-binding proteins. The mutation is located in the B site which is responsible for ATP binding. This protein probably provides energy to the xylose and disaccharide transport system as a result of the hydrolysis of ATP. PMID- 7494487 TI - NodS is an S-adenosyl-L-methionine-dependent methyltransferase that methylates chitooligosaccharides deacetylated at the non-reducing end. AB - In response to phenolic compounds exuded by the host plant, symbiotic Rhizobium bacteria produce signal molecules (Nod factors), consisting of lipochitooligosaccharides with strain-specific substitutions. In Azorhizobium caulinodans strain ORS571 these modifications are an O-arabinosyl group, an O carbamoyl group, and an N-methyl group. Several lines of evidence indicate that the nodS gene located in the nodABCSUIJ operon is implicated in the methylation of Nod factors. Previously we have shown that NodS is an S-adenosyl-L-methionine (SAM)-binding protein, essential for the L-[3H-methyl]-methionine labelling of ORS571 Nod factors in vivo. Here, we present an in vitro assay showing that NodS from either A. caulinodans or Rhizobium species NGR234 methylates end deacetylated chitooligosaccharides, using [3H-methyl]-SAM as a methyl donor. The enzymatic and SAM-binding activity were correlated with the nodS gene and localized within the soluble protein fraction. The A. caulinodans nodS gene was expressed in Escherichia coli and a glutathione-S-transferase-NodS fusion protein purified. This protein bound SAM and could methylate end-deacetylated chitooligosaccharides, but could not fully methylate acetylated chitooligosaccharides or unmethylated lipo-chitooligosaccharides. These data implicate that the methylation step in the biosynthesis pathway of ORS571 Nod factors occurs after deacetylation and prior to acylation of the chitooligosaccharides. PMID- 7494488 TI - An archaebacterial elF-1A: new grist for the mill. PMID- 7494486 TI - The leucine zipper of Aeromonas salmonicida AbcA is required for the transcriptional activation of the P2 promoter of the surface-layer structural gene, vapA, in Escherichia coli. AB - The Aeromonas salmonicida AbcA protein is involved in the synthesis of the O polysaccharide side-chains on the lipopolysaccharide and is also capable of enhancing the expression of the structural gene for the A-layer, vapA, when cloned into Escherichia coli. The P2 promoter of the vapA gene of A. salmonicida was cloned into a promoter probe vector and expression in E. coli was monitored. The expression of P2::lacZ was shown to be increased when abcA was provided in trans. AbcA contains an N-terminal ATP-binding domain as well as a C-terminal leucine zipper domain. Site-directed mutagenesis has been used to show that the ATP-binding domain is required for the synthesis of the O-polysaccharide side chains, but not for the enhancement of vapA expression. Conversely, the leucine zipper is needed for the increase in vapA expression, but not for O polysaccharide side-chain synthesis. This indicates that AbcA is a bifunctional protein that can influence the synthesis of the two principle antigenic components of the A. salmonicida cell surface. PMID- 7494489 TI - The malleability of the carboxy-terminus of the Escherichia coli beta' subunit of RNA polymerase: a bifunctional fusion to the LacZ(alpha) region. PMID- 7494490 TI - Molecular evolution of MRSA. PMID- 7494491 TI - Detection of methicillin-resistant Staphylococcus aureus (MRSA) with antibodies against synthetic peptides derived from penicillin-binding protein 2'. AB - Ten kinds of peptides (21 to 32 amino acids in length) were synthesized based on the reported amino acid sequences of the penicillin-binding protein 2' (PBP2') of methicillin-resistant Staphylococcus aureus (MRSA). Antibodies against these synthetic peptides (SPs) were generated by immunizing rabbits. The antibodies raised against all the peptides except for one reacted to PBP2' of MRSA and to SPs used for immunization but not to any other protein of MRSA or methicillin susceptible S. aureus (MSSA) tested by ELISA and Western blotting. A sandwich immunoradiometric assay (IRMA) for the detection of PBP2' was developed using these antibodies. The method could detect PBP2' extracted from as few as 3 x 10(4) cells of a clinical MRSA isolate, and a good correlation between cell number and signal radio-count was observed. IRMA was positive for all 51 methicillin-resistant staphylococci isolated from patients, and was negative for all the 28 methicillin-susceptible ones and 19 strains of other bacterial species. IRMA could be a simple and reliable method for MRSA detection in the clinical bacterial laboratory. PMID- 7494492 TI - Further characterization of Chlamydia pneumoniae specific monoclonal antibodies. AB - Studies using monoclonal antibodies have demonstrated species-specific reactivities with Chlamydia pneumoniae. In this study, further characterization of C. pneumoniae specific monoclonal antibodies TT-205 and RR-402 and description of C. pneumoniae specific antibodies prepared against other isolates are presented. TT-205 and RR-402 were shown to neutralize infectivity. Neutralization in cell culture was specific and enhanced by complement. Attempts to characterize the reactive antigen by immunoblotting, immunoaffinity chromatography and radioimmunoprecipitation were unsuccessful, probably due to difficulties in solubilizing the immunoreactive epitope without denaturing it. Recognition of the determinant by the monoclonal antibodies is labile to physical and chemical treatments suggesting that the reactive epitope is conformational. PMID- 7494494 TI - Distribution of C22-, C24- and C26-alpha-unit-containing mycolic acid homologues in mycobacteria. AB - There are three mycolic acid homologues with C22-, C24- and C26-alpha-units in Mycobacterium. In order to reveal the composition and distribution of these homologues in each subclass and molecular species of mycolic acids and to compare them with the composition of constitutive non-polar fatty acids (free and bound forms), we have separated non-polar fatty acids and each subclass of mycolic acids from 21 mycobacterial species by thin-layer chromatography, and analyzed non-polar fatty acid methyl esters by gas chromatography (GC) and the cleavage products of methyl mycolate by pyrolysis GC. We further performed mass chromatographic analysis of trimethylsilyl (TMS) ether derivatives of mycolic acid methyl esters by monitoring [B-29]+ ions (loss of CHO from the alpha branched-chain structure of mycolic acids) of m/z 426, 454 and 482 which are attributed to C22-, C24- and C26-alpha-units of TMS ether derivatives of methyl mycolates, respectively, (Kaneda, K. et al, J. Clin. Microbiol. 24: 1060-1070, 1986). By pyrolysis GC, C22:0, C24:0 and C26:0 fatty acid methyl esters generated by the C2-C3 cleavage of C22-, C24- and C26-alpha-unit-containing mycolic acid methyl esters, respectively, were detected. Their proportion was almost the same among subclasses of mycolic acids in every Mycobacterium and also similar to the proportion of constitutive non-polar C22:0, C24:0 and C26:0 fatty acids. By mass chromatography, the composition and distribution of C22- and C24-alpha-unit containing homologues were revealed to be similar between alpha- and alpha' mycolic acids in every Mycobacterium.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7494493 TI - Segmented filamentous bacteria are indigenous intestinal bacteria that activate intraepithelial lymphocytes and induce MHC class II molecules and fucosyl asialo GM1 glycolipids on the small intestinal epithelial cells in the ex-germ-free mouse. AB - In ex-germ-free mice conventionalized by association with fecal microorganisms, the induction fo major histocompatibility complex class II molecules and fucosylation of asialo GM1 glycolipid occur in the small intestinal epithelial cells (IEC). The intestinal intraepithelial lymphocytes (IEL), especially alpha beta T-cell receptor-bearing ones, also remarkably expand and show cytolytic activity. In this study, we investigated the immunological and physiological characteristics of the small intestine induced by a kind of indigenous bacteria of the small intestine, segmental filamentous bacteria (SFB), among chloroform resistant intestinal bacteria. Monoassociation of SFB with germ-free mice was confirmed by the determination of the base sequences of polymerase chain reaction products of 16S rRNA genes of the fecal bacteria of these mice and in situ hybridization using fluorescein-labeled probes based on them. SFB increased the number of alpha beta TCR-bearing IEL and induced Thy-1 expression and cytolytic activity of IEL. The induction of MHC class II molecules and fucosyl asialo GM1 glycolipids and the increases in the mitotic activity and the ratio of the number of columnar cells to those of goblet cells also occurred in the small intestinal epithelial cells on monoassociation of these bacteria. SFB are important indigenous bacteria for the development of the mucosal architecture and immune system in the small intestine, at least in mice. PMID- 7494495 TI - Inactivation of Lactobacillus bacteriophage PL-1 by microwave irradiation. AB - The effect of microwave irradiation on the survival of bacteriophage PL-1, which is specific for Lactobacillus casei, was studied using a commercial 2,450 MHz microwave oven. The phages were inactivated by microwave irradiation according to almost first-order reaction kinetics. The rate of phage inactivation was not affected by the difference in the continuous or intermittent irradiation, nor by the concentrations of phages used, but was affected by the volume of phage suspensions, which prevented the loss of generated heat. Microwave irradiation of phage suspensions produced a number of ghost phages with empty heads, but fragmentation of the tail was hardly noticed. The breakage of phage genome DNA was primarily caused by the heat generated by microwave irradiation, whereas the phage DNA was not affected by the same temperature achieved by heat from outside. Thus we concluded that the phage-inactivating effect of microwave irradiation was mainly attributed to a thermal microwave effect, which was much stronger than a simple thermal exposure. PMID- 7494496 TI - Seasonal distribution of adenoviruses, enteroviruses and reoviruses in urban river water. AB - In the 63-month period from January 1988 to March 1993, monthly levels of adenoviruses, enteroviruses (coxsackie B, polio, echo) and reoviruses in the urban river water in Nara Prefecture, Japan were in the range 0-25, 0-190 and 0 325, plaque forming units per liter (PFU/liter), and the average levels were 2.4, 40.6 and 56.2 PFU/liter, respectively. The peak reovirus level was found in winter during the cold weather months (Nov. to Mar.). The peak enterovirus level was found in summer (May to Sept.) but continued to be found in autumn-winter (Oct. to Jan.) from 1991 to 1993. The levels of adenoviruses were low throughout all 5 years, as compared to those of reoviruses and enteroviruses. Polioviruses were isolated following the administration of vaccine. Although a changing pattern of serotype prevalence was seen with the coxsackie B viruses and echoviruses from 1988 to 1993, this is not so for polioviruses, which remained almost unchanged for the five-year period. Adenoviruses were isolated throughout all five years, though in small numbers. Reoviruses were isolated most frequently throughout five years. PMID- 7494497 TI - Sequences of E/NS1 gene junction from four dengue-2 viruses of northeastern Thailand and their evolutionary relationships with other dengue-2 viruses. AB - We determined the 240-nucleotide sequences of the E/NS1 gene junction of four dengue-2 viruses by the primer extension dideoxy chain termination method. These viruses were isolated from dengue patients with different clinical severities in Nakhon Phanom, Northeastern Thailand in 1993. The results were compared with the 52 published dengue-2 sequences of the same gene region. Sequence divergence of four new isolates varied from 4.17% to 5.42% compared with dengue-2 prototype New Guinea C strain whereas it varied from 5.42% to 6.67% and from 6.67% to 7.09% when compared with Jamaica 1409 strain and PR159/S1 strain, respectively. All nucleotide substitutions were found at the 3rd position of the codons which were silent mutations. All 56 isolates studied were classified into five genotypic groups by constructing the dendrogram. The results indicated that four new isolates from Northeastern Thailand belong to genotype II of dengue virus serotype 2, and were most closely related to prototype New Guinea C strain. We also observed the variation in nucleotide and amino acid sequences among clusters of isolates (Thailand-1980, Malaysia-1989 and Thailand-1993) which were obtained from the dengue patients with different clinical severities. The significance of these genetic differences have been discussed in terms of the possible correlation between genetic variability and virulence. PMID- 7494498 TI - Humoral immunity to commensal oral bacteria: quantitation, specificity and avidity of serum IgG and IgM antibodies reactive with Actinobacillus actinomycetemcomitans in children. AB - The levels, specificity and avidities of serum IgM and IgG antibodies reactive with Actinobacillus actinomycetemcomitans (Aa) serotypes a, b and c were determined in periodontally healthy (PH) children and compared with subjects with localized juvenile periodontitis (LJP). All PH children exhibited IgM and IgG Aa reactive antibodies whether or not Aa was detected subgingivally but the antibodies were not specific for Aa. In contrast, LJP sera contained high concentrations of IgM and IgG antibodies reactive with Aa that were largely specific for this bacterium. IgM and IgG antibodies in both PH and LJP subjects were of low avidity. With one exception, the avidities of IgG anti-Aa antibodies were significantly greater than those of IgM antibodies in both PH and LJP subjects. However, although the LJP subjects had as much as 115-fold more Aa reactive IgG antibody than did the PH subjects the avidities of their IgG antibodies were no greater than those of the PH group. The induction by the host of low-avidity antibodies, that are ineffective in immune elimination, may be a reason why commensal bacteria persist at mucosal surfaces and why persons with LJP fail to eliminate Aa from their periodontal pockets. PMID- 7494499 TI - Down-regulation of T-cell proliferation in response to soluble anti-CD3 antibodies through development of redirected cytolytic activity eliminating costimulatory cells. AB - CD4+ T-depleted spleen cells (CD8+ T cells) activated by anti-CD3 antibodies (aCD3) suppressed proliferation of CD8+ T-depleted spleen cells (CD4+ T cells) and fresh normal T cells in response to aCD3. Antigen-nonspecific cytolytic activity was induced in splenic CD8+ T cells by stimulation with aCD3 and showed the peak level on day 3, whereas cytolytic activity induced in CD4+ T cells was weak. Intact Ig but not F(ab')2 of aCD3 induced and mediated cytolytic activity. Correspondingly the cytolytic activity induced by aCD3 was directed against target cells bearing Ig-binding Fc-receptor activity and cytolysis was inhibited by the addition of free Ig into the assay system. We showed that aCD3-activated T cells carried a high level of aCD3 on their surface at the time after the peak proliferation when they attained high cytolytic activity. This raised the possibility that the anti-CD3-induced aCD3-redirected cytolytic activity eliminated Fc-receptor-bearing costimulatory cells in the culture for down regulation of the T-cell proliferation. This view was supported by partial restoration of anti-CD3-induced low responsiveness of CD8+ T cells by the addition of fresh costimulatory cells. These results suggested a new pathway of down-regulation of T-cell proliferation by aCD3-activated cytolytic CD8+ T cells. PMID- 7494500 TI - Antibody responses raised against a conformational V3 loop peptide of HIV-1. AB - The amino acid sequence of the principal neutralizing determinant (PND) of 224 cases of human immunodeficiency virus type 1 (HIV-1) was determined and the most frequently occurring sequence was used as a peptide antigen for studying virus specific antibody responses. In our present study, a linear peptide of the most frequent PND was first synthesized and then oxidized to create a disulfide bridged loop conformation. Then, in order to construct a macromolecular structure for the purpose of increasing antigenicity, the synthetic peptide was conjugated to a core peptide. We compared the immunogenicity of the disulfide-bridged loop PND peptide antigen (AG4) and the linear PND peptide antigen (AG5). After immunizing rabbits 5 and 6 times with both peptides, the results obtained using ELISA revealed that AG4 (conformational-loop type) was more capable of inducing a high titer of antigen-specific antibodies than was AG5 (linear type). Despite an amino acid sequence homology of 72%, a 1:8 dilution of serum raised against AG4 inhibited 81.9% of HIV-1IIIB-mediated cell fusion, suggesting that conformational V3 loop peptide is able to elicit an antibody response which is strongly HIV-1 specific. PMID- 7494501 TI - Auxiliary method for clonal identification of Staphylococcus aureus by protein band pattern of released proteins on SDS-polyacrylamide gel. AB - A supportive method for clonal identification of Staphylococcus aureus strains was devised. Culture supernatant obtained by cellophane surface culture was subjected to sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) without performing any concentration procedure prior to electrophoresis. The combined use of cellophane surface culture and SDS-PAGE was convenient for determining whether the strains belonged to the same clone or not when conducted in conjunction with other tests for bacteriological characterization. PMID- 7494502 TI - Monoclonal antibody developed against a hemolysin of Bacillus thuringiensis. AB - A total of five hybridoma cell lines that produced monoclonal antibodies (MAb) against a hemolysin (Bt-hemolysin) produced by Bacillus thuringiensis were established and characterized. All of these monoclonal antibodies reacted similarly not only to Bt-hemolysin but also to a hemolysin (Bc-hemolysin) produced by B. cereus, suggesting that the two hemolysins are immunologically indistinguishable. The MAb developed in this study was also successfully applied for rapid and simple purification of both Bt- and Bc-hemolysins by immunoaffinity column chromatography. The partial N-terminal amino acid sequence of the purified hemolysins was determined to be Ile-Glu-Gln-Thr. PMID- 7494504 TI - Alteration of bacteriolytic enzyme profile of Staphylococcus aureus during growth. AB - Profiles of cell-associated bacteriolytic activities and those in the culture supernatant of Staphylococcus aureus FDA209P at various stages of growth were analyzed using sodium dodecyl sulfate-polyacrylamide gels containing Micrococcus luteus or S. aureus. In the logarithmic growth phase, the cell-associated bacteriolytic activities extracted with Triton X-100 contained a number of bacteriolytic proteins, the profiles of which were similar to those we reported elsewhere (Sugai, M., Akiyama, T., Komatsuzawa, H., Miyake, Y., and Suginaka, H. (1990) J. Bacteriol., 172, 6494-6498). The proteins include P1, P2, P7, P9, PX, P13, P18 and other minor components. At the stationary growth phase, the bacteriolytic band-profile of the Triton X-100 extract changed dramatically. P1, P7 and P9 disappeared, and the other minor bands had markedly decreased band intensities. On the other hand, P2, PX, P13, and P18 retained their band intensities during the stationary growth phase. The band intensities of P7, P13, PX, and P18 increased in the supernatant during the logarithmic growth phase. These results indicated that the bacteriolytic band-profile changes during growth. PMID- 7494505 TI - Antiviral activity of trichothecene mycotoxins (deoxynivalenol, fusarenon-X, and nivalenol) against herpes simplex virus types 1 and 2. AB - The effect of trichothecene mycotoxins, deoxynivalenol (DON), fusarenon-X (FX) and nivalenol (NIV), on plaque formation of herpes simplex virus types 1 and 2 (HSV-1 and HSV-2) in HEp-2 cells was examined. The 50% effective concentrations (EC50) of DON, FX, and NIV for HSV-1 plaque formation were 160, 56, and 120 ng/ml, respectively. Those for HSV-2 plaque formation were 94, 26, and 50 ng/ml, respectively. These three mycotoxins showed about 2-fold higher selectivity to HSV-2 than to HSV-1. Plaque formation of HSV-1 was not inhibited with trichothecenes at concentrations completely inhibiting plaque formation when cells were treated during virus adsorption period or 15 hr before infection. These results indicate that trichothecenes affect replication of HSV-1 after virus adsorption, but not before or during virus adsorption to the host cells. PMID- 7494503 TI - Functional activities of antibodies directed against surface lipoproteins of Borrelia hermsii. AB - Enriched preparations for mouse polyclonal immunoglobulin G (IgG) antibodies reactive with surface-exposed epitopes (Ab-SEE) of the 22-kDa and 24-kDa membrane lipoproteins of living Borrelia hermsii (HS 1 strain) cells were obtained by an antibody absorption technique using living spirochetes. In vitro, the antibody preparations both inhibited spirochetal growth and were borreliacidal in the presence of complement. The monovalent Fab antibody fragments, prepared from antibody-enriched preparations, did not inhibit the growth of the bacteria, whereas they killed the bacteria in the presence of complement. The two-dimension gel electrophoresis of B. hermsii cells showed that 3H-labeled fatty acids incorporated into the 22-kDa and 24-kDa lipoproteins were resolved into one and three compact spots, respectively. The spots were recognized by the Ab-SEE preparations reactive with the 22-kDa and 24-kDa proteins, by Western blotting. PMID- 7494506 TI - Childhood medulloblastoma in Ontario, 1977-1987: population-based results. AB - A retrospective review was carried out to study children, not more than 16 years old, with a confirmed diagnosis of medulloblastoma, who were residents of the Province of Ontario at the time of diagnosis between 1977 and 1987 inclusive. The provincial tumour registry provided the population database. One hundred and eight children with medulloblastoma were identified of whom 72 (67%) were initially treated at University of Toronto Centres and 36 (33%) at other Health Science Centres, hospitals, and Regional Cancer Centres (RCC) in Ontario. The hospital/Cancer Centre records were reviewed. The 5-year relapse-free survival (RFS) for all patients treated in Ontario was 58% (SE = 5%). Those treated in Toronto had a 5-year RFS of 65% (SE = 6%) compared to 44% (SE = 8%) for those treated in other RCCs in the province (P = 0.02). Relapse-free survival for the RCCs ranged from 25 to 60%, with a trend for improved survival with increasing centre size. Univariate analysis of determinants of relapse-free survival for all 108 patients showed the following variables to be significant: T-stage (Tx + T1 + T2 vs. T3A + T3B) P = 0.0004, M-stage (M0 + Mx vs. M1-4) P = 0.0006, extent of resection (total vs. less than total) P = 0.002, radiotherapy (craniospinal irradiation and posterior fossa boost vs. other) P = 0.02, and treatment centre (Toronto centres vs. RCC) P = 0.02. Cases treated at centres outside metropolitan Toronto had a nearly two-fold (relative risk = 1.93; 95% confidence interval = 1.07, 3.47) greater risk of recurrence or death than those seen in Toronto. However, in multivariate analysis this difference was not quite significant (P = 0.07) after controlling for stage (T and M), extent of resection, meningitis, and gender. These data suggest that patients with medulloblastoma should be referred for treatment to large centres with major pediatric neurosurgical and oncology resources. PMID- 7494508 TI - Cryopreservation of semen from adolescent patients with malignancies. AB - In adult oncological patients semen cryopreservation offers the possibility of preserving fertility prior to aggressive therapy that may lead to infertility. The cryopreserved semen can later be used to induce pregnancies in the partner by techniques of assisted fertilization. In adolescent boys the question of fertility is often beyond consideration when the young patient's life is threatened acutely. However, improved survival rates increasingly prompt the question of quality of life after therapy, including fertility. Semen quality is known to be impaired in patients with malignancies and may be further impaired by the process of cryopreservation. Since normal values for semen in adolescents are not known and spermatogenesis may be impaired by the malignant disease, it was unclear whether semen samples from adolescents with malignancies warrant cryopreservation at all. In order to demonstrate the feasibility of semen cryopreservation in adolescent males, we compared the results from 12 pubertal boys aged 14-17 years with those from 17 young adults aged 18-20 years who had similar malignancies and, additionally, to 210 adults with malignancies (> 20 years). Luteinizing hormone serum values were significantly lower in adolescents than in adult patients. Follicle stimulating hormone showed a significant increase with age. Testosterone serum levels and testicular volumes showed similar distribution patterns in adolescent and adult men. Sperm concentrations, sperm motility, and normal sperm morphology in the adolescent patients did not show significant differences compared with adults. Thus cryopreservation of semen should be considered as an option to young male patients whose cancer therapy will include potentially gonadotoxic treatment. PMID- 7494507 TI - Treatment of high-risk acute lymphoblastic leukemia in children using the AL851 and ALHR88 protocols: a report from the Kyushu-Yamaguchi Children's Cancer Study Group in Japan. AB - A total of 125 children, who were diagnosed as having high-risk acute lymphoblastic leukemia (ALL), were treated with two consecutive protocols designated as AL851 (1985-1988) and ALHR88 (1988-1990). All patients received induction therapy consisting of vincristine (VCR), prednisolone (PSL), daunorubicin (DNR), and I-asparaginase (I-Asp). In the ALHR88 protocol, the patients whose blasts in the bone marrow (BM) were > or = 25% on day 14 of induction therapy and who were classified into T-cell type received additional cytosine arabinoside (AraC). After consolidation with intermediate-dose methotrexate (MTX), reinduction therapy including VCR, dexamethasone, and adriamycin followed by high-dose AraC was done for all patients. Intrathecal MTX and 24Gy of cranial irradiation were used to prevent central nervous system leukemia. A maintenance therapy consisting of 6-mercaptopurine, cyclophosphamide, MTX, DNR, VCR, and AraC was administered for 3 years after achieving a complete remission (CR). CR was achieved in 51/55 (92.7%) for AL851 and 68/70 (97.1%) for ALHR88. The 5-year event-free survival rates were 49.1 +/- 6.7% in AL851 and 62.5 +/- 6.1% in ALHR88. The factors related to a poor prognosis were a high initial leukocyte count of greater than 50 x 10(9)/L (P < 0.001), an L2 morphology of leukemic cells by FAB classification (P = 0.009), the chromosomal abnormality (P = 0.004) and high residual leukemic cells in BM (> or = 25%) on day 14 of induction therapy (P < 0.001). Taking these factors into consideration, more intensive protocols were started in 1990 for the patients with high-risk ALL. PMID- 7494509 TI - Nonlymphoid gastrointestinal malignancies in Turkish children. AB - Nonlymphoid gastrointestinal malignancies are not frequently encountered malignant tumors of childhood. From 1972 to 1994, at Hacettepe University Children's Hospital, there were 6,774 malignant tumors encountered in patients from birth to age 17. Thirty-five of them (0.5%) originated from the gastrointestinal tract. There were five carcinoid tumors, four pancreas tumors (two were solid-cystic tumors), four APUD-omas, three leiomyosarcomas, one mesothelioma, two carcinomas of the stomach and duodenum, and 16 colon and rectum carcinomas (12 were mucin-producing). These rare tumors are discussed according to the characteristics of the patients, therapy modalities applied, and outcome of the patients. PMID- 7494510 TI - Ifosfamide tolerance in osteosarcoma patients previously treated with cis diamminedichloroplatinum-II: renal, hematologic, and neurologic observations. AB - We attempted to ascertain renal, hematologic, and neurologic tolerance to ifosfamide (IFX) in pediatric patients previously treated with large single and cumulative doses of cis-Diamminedichloroplatinum-II (CDP) for osteosarcoma (OS). Twenty OS patients were treated with CDP: initially 150 mg/m2 was administered every 2 weeks for a maximum of seven courses. Later, other agents, including additional CDP, were also administered. Twelve patients were treated with intra arterial CDP, one with intra-arterial, and later intravenous CDP, and seven with intravenous CDP. Patients who relapsed were treated with IFX. Renal function was monitored by measuring creatinine clearance, serum electrolytes, total protein, albumin and CO2 content, and urine analysis during IFX therapy. Prior to initiation of IFX, creatinine clearance was above 60 ml/min/m2 in all except one patient who had developed a hemolytic uremic syndrome (HUS). Cumulative CDP doses ranged from 300 to 22,500 mg/m2, and cumulative IFX doses 12 to 128 gm/m2. Myelosuppression was monitored by obtaining routine hemograms midway between each course of treatment. Neurologic tolerance was assessed by reviewing the medical records for any abnormality. The interval between CDP and IFX ranged from 1 to 64 months. All patients experienced a progressive reduction in creatinine clearance with CDP. The reduction in creatinine clearance, measured from base-line after three to four courses varied from 10 to 53.7%, after four to seven courses from 19 to 78%, and after seven courses from 12 to 80.5%. In all patients except five, including the HUS patient, creatinine clearance remained above 60 ml/min/m2 during IFX therapy. Twelve patients developed hypo-magnesemia in the vicinity of 1.4 to 1.6 mg/dl during CDP treatment and required magnesium supplementation. They were asymptomatic and the abnormality did not affect IFX tolerance. Fourteen patients intermittently displayed variable degrees of glycosuria, phosphaturia, and/or proteinuria during IFX therapy. This was considered to be a forma frustre type of Fanconi's syndrome. Approximately 80% of courses of IFX were associated with reversible myelosuppression. No neurologic abnormalities were detected. The abnormalities detected during IFX treatment were not major, did not give rise to symptomatology, and did not require discontinuation of therapy. Renal abnormalities were considered a forma frustre type of Fanconi's syndrome. Provided a creatinine clearance of 60 ml/min/m2 is accepted as a prerequisite for treatment, and no major preexisting renal disease is present, IFX is well tolerated by most patients previously exposed to very high cumulative doses of CDP. PMID- 7494511 TI - Elevated levels of tumor necrosis factor-beta, gamma-interferon, and IL-6 mRNA in Castleman's disease. AB - Castleman's disease (CD) is a lymphoproliferative disorder characterized by enlarged hyperplastic lymph nodes. CD may be localized or multifocal, and is often associated with signs and symptoms of generalized inflammation. The systemic manifestations of CD have been previously attributed to an overproduction of interleukin-6 (IL-6) by the tumor, although there is evidence that IL-6 is not responsible for all of the symptoms. We describe a 9-year-old boy who developed Castleman's disease with systemic findings of hypochromic microcytic anemia, growth arrest, inflammation, and hyperimmunoglobulinemia. Following surgical resection, all of the symptoms and laboratory abnormalities resolved. Using reverse transcriptase polymerase chain reaction (RT-PCR) analysis of the tumor, we found elevated levels of IL-6 mRNA as expected, but also elevated levels of tumor necrosis factor beta (TNF-beta) and gamma interferon (gamma-IFN) mRNA. Because these cytokines are mediators of immune regulation and inflammation, we propose that TNF-beta and gamma-IFN also play an important role in the pathophysiology of Castleman's disease. PMID- 7494512 TI - Familial sideroblastic anemia with emergence of monosomy 5 and myelodysplastic syndrome. AB - The case history of two sisters with pyridoxine-refractory familial sideroblastic anemia (FSA) is presented in which one developed a myelodysplastic syndrome (MDS) with monosomy for chromosome 5. Bone marrow examination of both patients at diagnosis showed erythroid hyperplasia with more than 50% ring sideroblasts. Karyotypic analysis initially showed a normal 46, XX karyotype in both of the children. Therapeutic trials with pyridoxine, prednisone, and erythropoietin were unsuccessful. The first patient required regular transfusions and developed a significant hemosiderosis. At the age of 9 years, 7.5 years after the diagnosis of FSA, refractory anemia with excess of blasts (RAEB) was diagnosed. Bone marrow cytogenetic analysis revealed a clone with monosomy for chromosome 5. Her sister's illness was detected at the age of 12 years. She has a more benign course of disease, remains largely transfusion independent and until now shows no signs of myelodysplasia. To our knowledge this is the first observation of a transition of FSA to MDS accompanied by the appearance of a chromosomal abnormality. FSA might be another type of bone marrow failure syndrome, therefore close follow-up of these patients may be necessary. PMID- 7494513 TI - Urinary excretion of methylated catecholamine metabolites in a child with neuroblastoma maturing into ganglioneuroma. AB - Neuroblastomas are malignant tumors derived embryonically from the neural crest. Biological diagnosis relies on assay of urinary excretion of homovanillic acid (HVA), vanillylmandelic acid (VMA), and dopamine (DA). Spontaneous regression of these neoplasms has been reported by numerous investigators. The authors report the case of a child with neuroblastoma that illustrates the relationship between catecholamine metabolites and tumor maturation. At 1 month of age, this infant presented an adrenal neuroblastoma with multiple metastases (stage IV); the initial histological diagnosis based on examination of cutaneous metastases was neuroblastoma. At the age of 6 months, after chemotherapy, the primary tumor was resected; hepatic metastases were discovered at laparotomy. The histological diagnosis for all lesions was highly differentiated, mature ganglioneuroma-like tissue. The main biochemical abnormality at the time of diagnosis was an elevation in normetanephrine (NMN). HVA was only slightly increased but rose progressively during chemotherapy; it dropped back to normal levels after the sixth course. This case illustrates the potential benefits of separate assays of urinary methylated catecholamine metabolites for biochemical diagnosis and therapeutic management of neuroblastoma in addition to assays of HVA, VMA, and DA. Case findings suggest existence of a transformation process with maturation of the tumor involving enzymatic regulation and expression of MAO. PMID- 7494514 TI - Juvenile fibrosarcoma of the temporal bone. AB - A case of juvenile fibrosarcoma arising from the head and neck region is described. This type of tumour should be considered as a separate entity different from the fibrosarcoma in adults because of the different clinical behaviour. The symptomatology, the radiographic features and the literature data are reviewed. PMID- 7494515 TI - Child with Wilms' tumor and von Willebrand disease at diagnosis and apparent complete response to chemotherapy after multiple relapses. PMID- 7494516 TI - Cytomegalovirus causing pericarditis with tamponade in an adolescent with cancer. PMID- 7494517 TI - Late local recurrence of Wilms' tumor. PMID- 7494518 TI - Factors that influence the further survival of patients who survive five years after the diagnosis of cancer in childhood or adolescence. PMID- 7494519 TI - Alkali therapy, hyperphosphatemia, and acute tumor lysis syndrome. PMID- 7494520 TI - Hepatoma in the elderly--follow-up on a case report. PMID- 7494521 TI - Ask the nurse attorney. PMID- 7494522 TI - An ethical practice model in the age of empowerment. PMID- 7494523 TI - An uncompromising view of nurse education, England, 1995. PMID- 7494524 TI - Health and the curriculum: an illuminative evaluation--Part 2: Findings and recommendations. PMID- 7494525 TI - Is disability any business of nurse education? AB - Perceptions of and assumptions about disability as diagnosis-based, individualistic, and necessarily tragic, still appear in the nursing literature as well as the general media. The rejection of a medical model of disability by writers who are themselves disabled, and the construction of an alternative social model, might appear to leave nurses--as professional interventionists- without any role in the care and support of physically disabled people. Such a view, as expressed by non-nurse writers, fails to acknowledge the fundamental changes that have occurred in both nursing and nurse education. This article criticizes the presently politically correct social model of disability, and describes an alternative 'interface model', which centres on the actual daily experience of disabled people. This model suggests that there is still a positive and welcome role, as informed partner, for the nurse and his or her disabled client. PMID- 7494526 TI - Measurement of the professional self-concept of nurses: developing a measurement instrument. AB - The professional self-concept of nurses is an important component in the professional and academic development of the nursing profession. Few studies have concentrated on the careful development of the construct of professional self concept in nurses, using a nursing sample, and particularly an Australian sample. This study focuses on a pilot group of nurses from the Hunter Region of New South Wales to develop the multi-dimensional construct professional self concept of nurses. A 56 item instrument using Likert scales was developed to measure the constructs flexibility/creativity, knowledge, skill/competence, caring, communication, leadership and satisfaction. The instrument was administered to 170 third year students enrolled in a Diploma of Applied Science (Nursing) and the results subjected to item analysis, factor analytical studies, and reliability estimates. Three factors emerged which accounted for 40% of the variance in responses constituting the professional self-concept of nurses: 'professional practice' (incorporating the dimensions leadership, skill and flexibility), 'satisfaction', and 'communication'. The study contains suggestions for the further use and research implications for a 27 item instrument for measuring the professional self-concept of nurses which has demonstrated validity and reliability. PMID- 7494527 TI - Education for change in a post-modern world: redefining revolution. AB - Social and behavioural sciences are established components in the curriculum of undergraduate nursing degrees. The purpose is to introduce future and practising nurses to the social and political influences which inform their workplaces and practices. Inevitably an awareness of the structural barriers and the powerful political interests involved in health can lead to feelings of powerlessness and despair of achieving change. Yet the skills of critical analysis and political awareness developed in study such as this are essential for health workers in the increasingly complex and politically charged domain in which they work. This paper will explore problems and barriers encountered in development of curriculum and teaching social and behavioural sciences in health. It will propose an alternative conceptual model, based on post-structuralism, as one way of addressing these barriers. This approach shifts the focus from meta-theoretical sociological concepts such as class, gender and culture, to one of examining subject positions, discourse, contestation and local action, thus enabling the exploration and development of possibilities for change. The paper will also provide a case study to illustrate this alternative approach. PMID- 7494528 TI - The 'political correctness' debate and caring in psychiatric nursing. AB - In recent years the controversy over so-called 'political correctness' has figured prominently in discourses of higher education. In terms of nursing, the issue of 'political correctness' cannot be confined to intellectual word-games, but is of key significance in the debate around the nature of professional caring. This paper is a discussion of the relevance of the controversy around 'political correctness' to the practice of psychiatric nursing. It placed the 'political correctness' debate in the context of the current debate within nursing about definitions of 'caring', and discusses the connections between both these debates and the theoretical imperatives of person-centered psychology. After discussing the particular problems which arise when the three issues of 'political correctness', 'caring' and humanistic psychology are brought together in this way; it proposes a politically focused strategy for the future development of psychiatric nursing. Although it is written from the perspective of psychiatric nursing in Britain and is very much concerned with the use of a particular language; the arguments put forward are equally appropriate to other English-speaking countries, and perhaps also to non-English speaking countries. PMID- 7494529 TI - The nurse teacher's role in the promotion of reflective practice. AB - The art of reflection is seen as a key process in bringing together practice and knowledge in nursing. More specifically, reflection addresses the artistic elements of nursing care delivery. It thus appears a useful skill for pre registration students to begin developing. However, research suggests that students under the age of 25 may lack both the cognitive readiness and the experience necessary for mature critical reflection. Teachers must therefore examine whether the reflective strategies and models that they present meet these students educational needs. It is postulated that simple models have a greater chance of success than those that are complex and demand a high time commitment. One such model for reflective journal writing is suggested. PMID- 7494530 TI - Essay writing in nursing: alerting students and teachers to the educational benefits. AB - Essay writing has recently emerged as an important tool in the appraisal of nursing students at pre- and post-registration level. However, despite its popularity, this activity seems to be somewhat taken for granted. Undoubtedly some nursing students find the transition to essay writing in Higher Education difficult. They seem unaware of assessment procedures, the importance of feedback and more importantly the individual sub-processes which go to make up the process of writing itself. Consequently, the essay is seen as a task and not as an opportunity for learning. This paper is aimed at pre- and post-registration nursing students and their teachers. The paper seeks to reveal some of the research evidence around essay writing. Under the heading of research, four areas are explored. These include the context of essay writing; writing itself; essay planning and finally perspectives on essay writing. The implications of this research are then examined under the separate headings of assessment procedures and the nature of teachers' feedback comments. ... The Essay then, is not a routine exercise, a chore for the student to write, a chore for the professor to read. It is, if properly approached by both parties, education in the truest sense. (Harris & McDougall 1958, p viii) PMID- 7494531 TI - Students' evolving beliefs about nursing: from entry to graduation in a four-year baccalaureate programme. AB - Students become socialised into nursing during an educational programme through interaction with faculty members, classes and seminars, and by the experience of practicing nursing. In this paper, some results from a qualitative study that examined the socialisation of nursing students from one four-year baccalaureate programme are described. The changes in the students' attitudes and values are presented. Davis' (1975) theory of doctrinal conversion is used to examine progress in the process of socialisation. There is evidence that students moved from a lay to a professional image of nursing over the course of their four-year programme. While they tended to remain idealistic, elements of realism were evident in the final year as they looked toward their role as a registered nurse and recognised the constraints inherent in the hospital system. PMID- 7494532 TI - Story telling: a search for meaning within nursing practice. AB - Story telling has been used for centuries as a powerful vehicle for communication, education, recreation and the preservation of cultural identity. In this paper the appeal of stories is discussed, and placed in a nursing context, with reference to the health care environment, which is increasingly dominated by technology, and the collection of impersonal 'information' against a background of market forces and competition. It is argued that this environment alienates nurses from each other, and from their patients, leaving them emotionally impoverished and distanced from the basic humanity of their craft. The utility of storytelling as a professional development tool is discussed, and numerous examples reviewed, in relation to pre-registration, post registration and ongoing experiential learning situations. The simplicity and immediacy of the storytelling tradition is compared to the academically generated and rhetoric laden notion of 'reflective practice'. The paper concludes that storytelling is an accessible yet powerful tool which contextualises and humanises nursing knowledge, facilitating a deeper understanding of self and others. PMID- 7494533 TI - The use of posters in the teaching of biological sciences on an undergraduate nursing course. AB - Posters are commonly used at research conferences for communicating information and in education as an alternative way of teaching and assessing students. We report the use of posters as a means of teaching and assessing part of the biological sciences component of an undergraduate nursing course. Tutors felt it was a valuable exercise enabling students to develop a number of skills, alongside the learning and revision of knowledge, including working as a team, condensing information and word and graphic processing. Students evaluated the exercise as being challenging but rewarding and preferred to be assessed by this method than by oral presentations or examinations. Lecturers and guided study/tutorials were preferred to posters as teaching methods. PMID- 7494534 TI - Integrating primary and secondary care--continuing education for acute and community nurses. AB - The current emphasis on shorter hospital stays and increased care in the community has led to a greater focus on integration of patient care between primary and secondary health care settings. The module which is discussed in this paper aims to address the knowledge gaps of trained nurses, enabling them to understand current changes in health care in order to enhance their practice. Flexibility in pre-registration education allows for incorporation of this new knowledge as changes occur. A situational analysis approach was used to inform the curriculum, and wide consultation took place with service providers in both hospital and community. The resulting first module addresses care in the community, influences on patient/client's health experience, current social policy and local initiatives in response to legislation, and role of carers. Assessment takes the form of a client profile. To date evaluation of the module has been extremely positive and the curriculum planning team are developing a more practice focused second module which will address skills It is hoped that knowledge gained from both modules will enhance practice and form an initial preparation for secondary care nurses wishing to move into the community. PMID- 7494535 TI - Political principles in action: a critical analysis of Education and Training: Working Paper 10 and its implications for nurse education. AB - Education and Training: Working Paper Ten (WP10) (Department of Health 1989a) was published in October 1989. It was an integral part of Working for Patients (Department of Health 1989b) and as such was alleged to embrace the same fundamental principles. WP10 may result in the most significant change to nurse education in the UK since the 1949 Nurses Act, with variations in its funding, structure and organisation. Surprisingly, there has been relatively little discussion about this important development within the nursing press. The aim, therefore, of this paper is to evaluate WP10 within the context of the ideology which has underpinned social policy in the last 14 years. Firstly, Conservative Party policy will be explored, the WP10 itself will then be examined in order to assess how far such principles have been applied to nurse education through its implementation. It is intended that by this process of analysis further understanding will be gained about this policy and its potential consequences for the future of nursing. PMID- 7494536 TI - Healthcare changes bring increased liability risk for nurses. PMID- 7494537 TI - Compared to what? PMID- 7494538 TI - Response of the rat heart to catecholamines and thyroid hormones. AB - Catecholamines and thyroid hormones have a similar influence on heart function and metabolism, but this may occur in a differential manner and to a different extent. In this study, the effects of norepinephrine (NE) and of triiodothyronine (T3) were studied in regard to the function of the left (LV) and right ventricle (RV) and to the oxidative pentose phosphate pathway (PPP). NE was applied in rats as continuous i.v. infusion (0.2 mg/kg/h) for three days. T3 was given as daily s.c. injections (0.2 mg/kg) for the same period of time. LV and RV function was measured in the closed-chest trapanal-anesthetized animals using special Millar ultraminature catheter pressure transducers. NE induced an increase in heart rate, in mean arterial pressure, and in total peripheral resistance (TPR). The cardiac RNA/DNA and the left ventricular weight/body weight ratios were increased by about 40%. These effects were prevented by simultaneous alpha- and beta receptor blockade with prazosin and metoprolol, respectively, but not by verapamil which abolished the hemodynamic effects. RVSP was significantly elevated by NE in a dose-dependent manner. The functional effects of T3 on the LV were not as pronounced as those induced by NE. Heart rate and LV dp/dtmax were increased by T3, and this increase was prevented by concomitant beta-receptor blockade with metoprolol. In contrast to NE, T3 induced an increase in cardiac output and a concomitant decrease in TPR. The RNA/DNA ratio was elevated and cardiac hypertrophy had developed after treatment for three days with T3. These changes were not affected by beta-receptor blockade with metoprolol. RVSP was increased by T3 to a lesser extent than with NE. In metabolic terms it turned out that only NE, but not T3 had a stimulating effect on the cardiac PPP. NE increased the mRNA and activity of glucose-6-phosphate dehydrogenase (G-6-PD), the first and regulating enzyme of this pathway. However, there was no effect of T3 on G-6-PD activity nor on 6-phosphogluconate dehydrogenase activity, one of the following enzymes in the pathway within the first 5 days of T3 treatment. These results demonstrate that the functional effects of T3 were not as pronounced as or even different from those of NE, and that T3 lacked a stimulating effect on the cardiac PPP. PMID- 7494539 TI - On the mechanism and possible therapeutic application of delayed adaptation of the heart to stress situations. AB - Mild (not harmful) stress may initiate an adaptive mechanism, protecting the heart from harmful consequences of a more severe stress. There are at least three known types of cardiac adaptation to stress, such as: a) the gradually developing, long lasting adaptation to chronic mechanical overload, leading to cardiac hypertrophy, later to cardiomyopathy and heart failure, b) the rapidly developing adaptation to moderate stress initiated by 'preconditioning' brief coronary occlusion(s) or brief periods of rapid cardiac pacing, protecting for less than 1 h against consequences of a subsequent, severe stress, c) the later appearing, more prolonged cardio-protective adaptation, described by us in 1983, induced by various forms of more severe but not injurious stimuli, such as an optimal dose of prostacyclin or its stable analogues; or a series of brief periods of rapid pacings. This form of cardiac adaptation to stress protects for 24-48 h against consequences of a more severe stress such as: 1. myocardial ischaemia; 2. early and late postocclusion and reperfusion arrhythmias; 3. early morphologic changes secondary to ischaemia and reperfusion; 4. ischaemia induced myocardial loss of K+ and accumulation of Na+ and Ca++; 5. it may increase the tolerance to the toxic effects of cardiac glycosides. A reduced response to beta adrenergic stimuli and a concomitant increase in activity and amount of PDE I and IV was shown by us earlier.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7494540 TI - Ischaemic preconditioning in the rat heart: the role of G-proteins and adrenergic stimulation. AB - Since recent findings indicate the involvement of G-proteins in the mechanism of ischaemic preconditioning (PC), the present study was aimed to investigate the role of adrenergic mechanisms, such as G-proteins and stimulation of adrenergic receptors, in this phenomenon. For this purpose, isolated Langendorff-perfused rat hearts were subjected to regional ischaemia (30 min occlusion of LAD) followed by reperfusion. The effect of PC (a single 5 min occlusion/reperfusion before a long occlusion) on ischaemia- and reperfusion induced arrhythmias was studied in conjunction with an assessment of G-proteins in the myocardial tissue by means of Western blotting and ADP-ribosylation with bacterial toxins. To follow the link between G-proteins and adrenergic receptors, their stimulation by exogenous norepinephrine (NE) was applied to test whether it can mimic the effect of PC on arrhythmias. Thirty min ischaemia and subsequent reperfusion induced high incidence of ventricular tachycardia (VT) and fibrillation (VF). PC significantly reduced a total number of extrasystoles, incidence of VT and abolished VF. It was, however, insufficient to suppress reperfusion-induced sustained VF. Measurement of G-proteins revealed that PC led to a reduction of stimulatory Gs proteins, whereas inhibitory Gi proteins were increased. NE (50 nmol) introduced a manner of similar to PC (5 min infusion, 10 min normal reperfusion) reduced ischaemic arrhythmias in the same way, as PC. In addition, in NE-pretreated hearts reperfusion induced mostly transient VF, which was spontaneously reverted to normal sinus rhythm. A transient increase in heart rate and perfusion pressure during NE infusion completely waned before the onset of ischaemia, indicating that antiarrhythmic effect was not related to haemodynamic changes and to conditions of myocardial perfusion. CONCLUSION: antiarrhythmic effect of PC may be mediated by a stimulation of adrenergic receptors coupled to appropriate G-proteins. Consequently, the inhibition of adenylate cyclase activity and reduction in cAMP level, as well as the activation of protein kinase C may be considered as two possible pathways leading to a final response. PMID- 7494542 TI - Importance of monocytes/macrophages and fibroblasts for healing of micronecroses in porcine myocardium. AB - In porcine heart, embolization of small coronary arteries with microspheres in 25 microns in diameter induces collateral capillary vessel growth by angiogenesis in and around focal necrosis. By histological analysis the inflammatory infiltrates in this porcine tissue were characterized by numerous monocytes/macrophages and fibroblasts as well as neutrophils and numerous capillaries, some in mitosis. The aim of the present study, therefore, was to clarify the role of monocytes/macrophages and fibroblasts in angiogenesis and in repair in ischemic porcine myocardium. Using a human acidic fibroblast growth factor (aFGF) cDNA probe for in situ hybridisation labeling for aFGF mRNA was seen in monocytes and macrophages only, beginning at day 1, with a maximum at 3 and 7 days, and minimal labeling at 4 weeks. We have also shown, with a specific antibody and fluorescence microscopy, that tumur necrosis factor alpha (TNF alpha) follows the same time sequence and that it is produced by monocytes/macrophages. The number of capillaries in infiltrates at 3 and 7 days as revealed by the lectin Dolichus Biflorus Agglutinin was high and declined at 4 weeks. In situ hybridisation using a rat cDNA probe for fibronectin showed the increased production of fibronectin mRNA in fibroblasts. To describe the expression of fibronectin and the collagens I, III, VI immunohistochemistry was used. A comparison showed that fibroblasts produced fibronectin mRNA starting at day 3, but the protein was only maximally expressed at day 7 and 4 weeks. Collagen I, III, VI expression was highest at 1-4 weeks.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7494541 TI - Adaptation of the heart to ischemia by preconditioning: effects on energy equilibrium, properties of sarcolemmal ATPases and release of cardioprotective proteins. AB - Ischemic preconditioning of the heart is referred as a manifest increase in tolerance of the myocardium to otherwise damaging ischemic insult, achieved by one or few consequent initial short exposures to ischemia, each followed by reperfusion of the ischemic area. Several mechanisms such as opening of collateral vessels, the action of catecholamines, inositol phosphates, G-proteins and/or adenosine; inhibition of mitochondrial ATPase, the effects of different endogenous protective substances like heat stress or shock proteins, etc., are believed to cooperate in the mechanism of induction of preconditioning or in maintaining its effect. The present study is an attempt to extend the present knowledge about preconditioning from two aspects: i.) the peculiarities of energy equilibrium in preconditioned myocardium including adaptation of cardiac sarcolemmal ATPases to ischemia and/or hypoxia, and ii) participation of a new endogenous cardioprotective substance in the mechanism of preconditioning. The energy equilibrium in preconditioning is characterized by adaptation of cardiac energy demands to the capacity of energy production and delivery decreased by anaerobiosis and is manifested by constant ratios between ATP, ADP, AMP and the sum of ADN. Principles are proposed that may enable a prediction and mathematical modelling of the balanced energetic state in the preconditioned myocardium. These principles are based on thermodynamics and involve besides others a more economic handling of ATP by sarcolemmal ATPases. The latter enzymes adapt themselves to lowered availability of ATP by decreasing besides their Vmax also their values of Km (increase in the affinity) for ATP and some of them even adjust their activation energy (the anaerobiosis-induced elevation of Ea.t. is missing).(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7494543 TI - Inhibition of cardiac sarcolemma Na(+)-K+ ATPase by oxyradical generating systems. AB - The Na(+)-K+ ATPase activity and SH group content were decreased whereas malondialdehyde (MDA) content was increased upon treating the porcine cardiac sarcolemma with xanthine plus xanthine oxidase, which is known to generate superoxide and other oxyradicals. Superoxide dismutase either alone or in combination with catalase and mannitol fully prevented changes in SH group content but the xanthine plus xanthine oxidase-induced depression in Na(+)-K+ ATPase activity as well as increase in MDA content were prevented partially. The Lineweaver-Burk plot analysis of the data for Na(+)-K+ ATPase activity in the presence of different concentrations of MgATP or Na+ revealed that the xanthine plus xanthine oxidase-induced depression in the enzyme activity was associated with a decrease in Vmax and an increase in Km for MgATP; however, Ka value for Na+ was decreased. Treatment of sarcolemma with H2O2 plus Fe2+, an hydroxyl and other radical generating system, increased MDA content but decreased both Na(+) K+ ATPase activity and SH group content; mannitol alone or in combination with catalase prevented changes in SH group content fully but the depression in Na(+) K+ ATPase activity and increase in MDA content were prevented partially. The depression in the enzyme activity by H2O2 plus Fe2+ was associated with a decrease in Vmax and an increase in Km for MgATP. These results indicate that the depressant effect of xanthine plus xanthine oxidase on sarcolemmal Na(+)-K+ ATPase may be due to the formation of superoxide, hydroxyl and other radicals. Furthermore, the oxyradical-induced depression in Na(+)-K+ ATPase may be due to the formation of superoxide, hydroxyl and other radicals.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7494544 TI - Alterations of beta-adrenoceptor-G-protein-regulated adenylyl cyclase in heart failure. AB - Alterations of receptor-G-protein-regulated adenylyl cyclase activity have been suggested to represent an important alteration leading to contractile dysfunction in the failing human heart. Recent experiments suggest that the beta 1 adrenoceptor (beta 1 AR) density and mRNA levels are reduced, while beta 2 adrenoceptors and stimulatory G-proteins are unchanged (mRNA and protein level). Functional assays demonstrated that the catalyst of the adenylyl cyclase is not different between failing and nonfailing myocardium. Inhibitory G-proteins are increased (pertussis toxin substrates, protein and mRNA) and correlate to the reduced inotropic effects of beta-adrenoceptor agonists and of cAMP-PDE inhibitors. Gi alpha-coupled m-cholinoceptors and A1-adrenergic receptors are unchanged in density and affinity. Stimulation of these receptors resulted in an unchanged antiadrenergic effect on force of contraction. In conclusion, a downregulation of beta 1 AR and an increase of Gi alpha have been observed as signal transduction alteration in failing human myocardium. These alterations are due to alterations of gene expression in the failing heart and are related to a defective regulation of force of contraction in heart failure. PMID- 7494545 TI - Immunocytochemical studies of the Gi protein mediated muscarinic receptor adenylyl cyclase system. AB - The localization of three key signal transduction components was indicated in rat heart tissue by immunocytochemical and histochemical experiment. It was shown that: 1. The M2 muscarinic receptors are localized along outer cell membranes and T-tubule membranes of cardiomyocytes but additionally at membranes of endothelial cells and fibroblasts. 2. Gia was found along outer cell membranes of cardiomyocytes and other cells of the heart and also inside the cells of the perinuclear space in close contact to the nuclei envelope and the endoplasmic reticulum membranes. Goa were found to be associated mainly in atrial tissue, especially at the nerval (neuronal) endings located among the cardiac muscle cells. This was shown in parallel incubation with specific neuronal antibody as a marker for these structures. 3. Adenylyl cyclase was localized along the sarcolemma and the T-tubule membranes in normal cardiomyocytes of rat and guinea pig hearts. Under ischemic conditions, the adenylyl cyclase was also seen in junctional sarcoplasmic reticulum membranes. The reasons for this changed localization need further elucidation. Binding of the adenylyl cyclase within the molecular structure of the membrane or variation of the marker penetration remain to be clarified. PMID- 7494547 TI - Effects of selective alpha 1A-adrenoceptor antagonists on reperfusion arrhythmias in isolated rat hearts. AB - Stimulation of alpha 1-adrenoceptors (AR) during ischaemia in the rat heart by exogenous phenylephrine exacerbates reperfusion arrhythmias, an effect apparently mediated by the alpha 1A-AR subtype. We tested whether alpha 1A-AR stimulation by endogenous catecholamines, released during ischaemia, could modulate reperfusion arrhythmias, using as pharmacological tools the selective alpha 1A-AR antagonists abanoquil (UK52046) and WB4101. Isolated rat hearts (n = 12/group) were subjected to dual coronary perfusion. After 15 min of aerobic perfusion of both coronary beds, abanoquil or WB4101 was infused selectively into the left coronary bed (LCB) for 5 min. The LCB was then subjected to 10 min of zero-flow ischaemia and 5 min of reperfusion. Effects on PR interval, width of the ventricular complex (QRST90) and reperfusion arrhythmias were assessed. Abanoquil at concentrations of 0.03, 0.1 and 0.3 microM tended to reduce the incidence of reperfusion-induced ventricular fibrillation (VF) in a dose-dependent manner from 75% in controls to 58, 33 and 25%, but this effects did not achieve statistical significance. Similarly, WB4101 at 0.1, 0.3 and 1 microM also tended to reduce VF incidence from 67% in controls to 67, 42% and 33% (NS). The incidence of ventricular tachycardia (VT) was 100% in all groups and ECG parameters were not altered significantly by either drug. These results suggest that, in this denervated isolated heart preparation, alpha 1A-AR stimulation during ischaemia by endogenous catecholamines does not significantly modulate reperfusion arrhythmias. PMID- 7494546 TI - Renaissance of cytochemical localization of membrane ATPases in the myocardium. AB - ATPases of cardiac cells are known to be among the most important enzymes to maintain the fluxes of vital cations by hydrolysis of the terminal high-energy phosphate of ATP. Biochemically the activities of Ca(2+)-pump ATPase, Ca2+/Mg(2+) ecto ATPase, Na+,K(+)-ATPase and Mg(2+)-ATPase are determined in homogenates and isolated membranes as well as in myofibrillar and mitochondrial fractions of various purities. Such techniques permit estimation of enzyme activities in vitro under optimal conditions without precise enzyme topography. On the other hand, cytochemical methods demonstrate enzyme activity in situ, but not under optimal conditions. Until recently several cytochemical methods have been employed for each enzyme in order to protect its specific activity and precise localization but the results are difficult to interpret. To obtain more consistent data from biochemical and cytochemical point of view, we modified cytochemical methods in which unified conditions for each ATPase were used. The fixative solution (1% paraformaldehyde-0.2% glutaraldehyde in 0.1 M Tris Base buffer, pH 7.4), the same cationic concentrations of basic components in the incubation medium (0.1 M Tris Base, 2 mM Pb(NO2)3, 5 mM MgSO4, 5 mM ATP) and selective stimulators or inhibitors were employed. The results reveal improved localization of Ca(2+)-pump ATPase, Na(+)-K+ ATPase and Ca2+/Mg(2+)-ecto ATPase in the cardiac membrane. PMID- 7494548 TI - The role of catecholamines on intercellular coupling, myocardial cell synchronization and self ventricular defibrillation. AB - Ventricular fibrillation (VF) is one of the most life threatening events. Although in humans VF is generally sustained (SVF) requiring artificial defibrillation, in various mammals and in some cases in humans VF terminates by itself, reverting spontaneously into sinus rhythm. Since VF is one of the main causes of sudden death, one of the important clinical problems today is if and how we can transform the fatal SVF into a self limited transient one (TVF). From electrophysiological studies carried out on anaesthetized open chest animals, we have found that TVF requires a high degree of intercellular coupling and synchronization. Cardiac myocytes are electrically coupled with adjacent cells. The intercellular coupling is a focus of low electrical resistance which allows rapid transmission of electrical impulses between cells. Any decrease in intercellular coupling decreases the ability of the heart for self defibrillation. The cell-to-cell coupling decreases with age, ischemia, VF and variations in physiological conditions probably due to an increase in intercellular resistance (Ri), widening in the internexal gaps, decrease in electrotonic space constant (lambda) etc. All of these factors are known to be affected by intracellular concentration of free Ca++ ([Ca++]). On the basis of studies carried out on various mammals at different ages, we hypothesized that the ability of the heart to defibrillate depends on the cardiac catecholamine level [CA], during VF. This hypothesis is supported by the facts, known from the literature, that increase in [CA] decreases intracellular free Ca++ concentration, decreases Ri and increases lambda.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7494549 TI - Inhibition of (Na/K)-ATPase by electrophilic substances: functional implications. AB - The effect of electrophilic substances: p-bromophenylisothiocyanate (PBITC); fluoresceinisothiocyanate (FITC); [4-isothiocyanatophenyl-(6-thioureidohexyl) carbamoylmethyl] -ATP (ATPITC); 2,4,6-trinitrobezenesulfonic acid (TNBS); 1-(5 nitro-2-furyl)-2-phenylsulfonyl-2-furylcarbonyl ethylene (FE1); 1-(5 phenylsulfonyl-2-furyl)-2-phenylsulfonyl-2-furylcarb onyl ethylene (FE2) and 1-(5 phenylsulfonyl-2-furyl)-2-phenylsulfonyl-2-tienocarb onyl ethylene (FE3) on the sarcolemmal (Na/K)-ATPase isolated from guinea-pig hearts was studied. FITC and PBITC were found to inhibit competitively the activation of (Na/K)-ATPase by ATP. Being for the enzyme inhibitor and substrate at the same time ATPITC does not offered clear kinetic behavior. However, the activation of (Na/K)-ATPase by sodium and potassium ions was inhibited non-competitively by all three isothiocyanates. These data indicated that isothiocyanates may interact predominantly in the ATP-binding site of the enzyme molecule. In contrary to isothiocyanates TNBS and FE1 (FE2 and FE3 were ineffective) inhibited the activation of (Na/K)-ATPase by ATP non-competitively i.e., their interaction in the ATP-binding site seemed to be improbable. Nevertheless, TNBS and FE1 both manifested affinities to that moiety of (Na/K)-ATPase molecule which is binding potassium. More specific was the effect of FE1 that showed clearly competitive inhibition of potassium-stimulation of the enzyme activity. FE1 exerted also an ouabain-like effect on the mechanical activity of isolated perfused guinea-pig heart. This result indicates that FE1 seems to exert a selective inhibition of the (Na/K)-ATPase not only in vitro but also in integrated cardiac tissue. PMID- 7494550 TI - Cell-cell and cell-matrix adhesion molecules in human heart and lung transplants. AB - The interaction of immune cells with endothelial and target cells and extracellular matrix in human organ transplants is regulated by a number of receptor-ligand molecules. The molecules mediating intercellular adhesion and activation are classified as integrin, immunoglobulin and selectin families. In the present study the patterns of their cellular expression in human heart and lung transplants are described in normal state and during transplant rejection. The results reveal an organ specific regulation of the different adhesion molecules during transplant rejection. Specific differences were noted in the endothelial expression of vascular ligand molecules in the vascular segments of heart and lung transplants, especially in the lung capillaries. Cell type specific patterns of intercellular and cell-matrix adhesion molecules as their ligands were found in different states of graft rejection. Intravascular and interstitial differences in the expression patterns of leukocyte adhesion receptors support a concept of their stepwise function during graft infiltration. The implications for the organ specific appearance of inflammatory reactions in human heart and lung transplants as for immunosuppressive therapy are discussed. PMID- 7494551 TI - Progression of heart failure: a role for interstitial fibrosis. AB - Progressive deterioration of left ventricular (LV) function is a characteristic feature of the heart failure (HF) state. The mechanism or mechanisms responsible for this hemodynamic deterioration are not known but may be related to progressive intrinsic dysfunction, degeneration and loss of viable cardiocytes. In the present study, we tested the hypothesis that accumulation of collagen in the cardiac interstitium (reactive interstitial fibrosis, RIF), known to occur in HF, results in reduced capillary density (CD = capillary/fiber ratio) and increased oxygen diffusion distance (ODD) which can lead to hypoxia and dysfunction of the collagen encircled myocyte. Studies were performed in LV tissue obtained from 10 dogs with chronic HF (LV ejection fraction 26 +/- 1%) produced by multiple sequential intracoronary microembolizations. In each dog, CD and ODD were evaluated in LV regions that manifested severe RIF (volume fraction 16 +/- 2%) and in LV regions of little or no RIF (volume fraction 4 +/- 1%). In regions of severe RIF, CD was significantly decreased compared to regions of no RIF (0.92 +/- 0.02 vs. 1.05 +/- 0.03) (P < 0.003). Similarly, ODD was significantly increased in regions of severe RIF compared to regions of no RIF (15.3 +/- 0.4 vs. 12.2 +/- 0.3 microns) (P < 0.001). These data suggest that in dogs with chronic HF, constituent myocytes of LV regions which manifest severe RIF may be subjected to chronic hypoxia; a condition that can adversely impact the function and viability of the collagen encircled cardiocyte. PMID- 7494552 TI - Chronic hibernating myocardium: interstitial changes. AB - Chronic left ventricular dysfunctional but viable myocardium of patients with chronic hibernation is characterized by structural changes, which consist of depletion of contractile elements, accumulation of glycogen, nuclear chromatin dispersion, depletion of sarcoplasmic reticulum and mitochondrial shape changes. These alterations are not reminiscent of degeneration but are interpreted as de differentiation of the cardiomyocytes. The above mentioned changes are accompanied by a marked increase in the interstitial space. The present study describes qualitative and quantitative changes in the cellular and non-cellular compartments of the interstitial space. In chronic hibernating myocardial segments the increased extracellular matrix is filled with large amounts of type I collagen, type III collagen and fibronectin. An increase in the number of vimentin-positive cells (endothelial cells and fibroblasts) compared with normal myocardium is seen throughout the extracellular matrix. The increase in interstitial tissue is considered as one of the main determinants responsible for the lack of immediate recovery of contractile function after restoration of the blood flow to the affected myocardial segments of patients with chronic left ventricular dysfunction. PMID- 7494554 TI - Role of early reperfusion in the induction of adhesion molecules and cytokines in previously ischemic myocardium. AB - Our studies in vitro demonstrate that neutrophil mediated injury of isolated cardiac myocytes requires the presence of ICAM-1 on the surface of the myocyte and CD11b/CD18 activation on the neutrophil. In post-ischemic cardiac lymph, there is rapid appearance of C5a activity during the first hours of reperfusion. Interleukin-6 activity is present throughout the first 72 h of reperfusion and is sufficient to induce ICAM-1 on the surface of the cardiac myocyte. In situ hybridization studies suggest that ICAM-1 mRNA is found in viable myocardial cells on the edge of the myocardial infarction within 1 h of reperfusion. ICAM-1 protein expression on cardiac myocytes is seen after 6 h of reperfusion, and increases thereafter. Non-ischemic tissue demonstrates no early induction of ICAM 1 mRNA or ICAM-1 protein on myocardial cells. In our most recent experiments, we have determined that reperfusion is an absolute requirement for the early induction of myocardial ICAM-1 mRNA in previously ischemic myocardial cells. To further assess this, we have cloned and sequenced a canine interleukin-6 (IL-6) cDNA. The data suggest that early induction of IL-6 mRNA is also reperfusion dependent as it could be demonstrated in the same ischemic and reperfused segments in which ICAM-1 mRNA was found. Peak expression of IL-6 mRNA occurred much earlier than that for ICAM-1 mRNA. Similar experiments were then performed with a molecular probe for interleukin-8 (IL-8). This chemokine is a potent neutrophil stimulant and has a higher degree of specificity for neutrophils than classic chemoattractants such as C5a.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7494555 TI - Ventricular remodeling: insights from pharmacologic interventions with angiotensin-converting enzyme inhibitors. AB - Structural remodeling of the left ventricular (LV) myocardium develops in a time dependent fashion following acute myocardial infarction and may be an integral component in the transition toward overt heart failure. Globally, the remodeling process is characterized by progressive LV enlargement and increased chamber sphericity. At the cellular level, the remodeling process is associated with myocyte slippage, hypertrophy, and accumulation of collagen in the interstitial compartment. In the present study, we examined the effects of early, long-term monotherapy with the angiotensin converting enzyme (ACE) inhibitor, enalapril, on the progression of LV remodeling in dogs with LV dysfunction (ejection fractions 30-40%) produced by multiple sequential intracoronary microembolizations. Dogs were randomized to 3 months oral therapy with enalapril (n = 7) or to no treatment (n = 7). In untreated dogs, LV end-systolic volume index (ESVI), end diastolic volume index (EDVI) and chamber sphericity increased significantly during the 3 months follow-up period. In contrast, in dogs treated with enalapril ESVI, EDVI and chamber sphericity remained essentially unchanged. Treatment with enalapril attenuated myocyte hypertrophy and the accumulation of interstitial collagen in comparison to untreated dogs. These data indicate that early treatment with ACE inhibitors can prevent the progression of LV remodeling in dogs with LV dysfunction. Afterload reduction, inhibition of direct action of angiotensin-II and possibly the decrease in bradykinin degradation elicited by ACE inhibition may act in concert in preventing the progression LV chamber remodeling. PMID- 7494553 TI - Differences between atrial and ventricular protein profiling in children with congenital heart disease. AB - The purpose of the present study was to compare protein profiling of atria and ventricles in children operated for congenital heart disease. Tissue samples were obtained during surgery from patients with normoxemic (ventricular and atrial septal defects) and hypoxemic (tetralogy of Fallot) diseases. Protein fractions were isolated by stepwise extraction from both right ventricular and atrial musculature. The concentration of total atrial protein in the normoxemic patients exceeded the ventricular value (110 +/- 2.1 vs 99.9 +/- 4.0 mg.g-1 wet weight, respectively); in the hypoxemic group this atrio-ventricular difference disappeared. The concentration of contractile proteins in all cardiac samples was significantly higher in the ventricles as compared with atria, while the concentration of collagenous proteins was significantly higher in the atria (due to a higher amount of the insoluble collagenous fraction). The concentration of sarcoplasmic proteins (containing predominantly enzyme systems for aerobic and anaerobic substrate utilization), however did not differ between ventricles and atria. Furthermore, ventricular contractile fractions obtained from both normoxemic and hypoxemic patients were contaminated with the myosin light chain of atrial origin. Soluble collagenous fractions (containing newly synthesized collagenous proteins, predominantly collagen I and III), derived from all ventricular samples, were contaminated by low molecular weight fragments (mol. weight 29-35 kDa). The proportion of the soluble collagenous fraction was significantly higher in atrial but not in ventricular myocardium of hypoxemic children as compared with the normoxemic group. It seems, therefore, that lower oxygen saturation affects the synthesis of collagen preferentially in atrial tissue. PMID- 7494557 TI - Biomechanical signals in the coronary artery triggering the metabolic processes during cardiac overload. AB - Peculiarities in structure and deformability of epicardial conduit coronary arteries are described. The thin wall of animal coronary artery contrasts the human coronary artery in which the remarkable wall thickness is due namely by the intima thickness. Deformation in length and diameter of conduit coronary arteries, due to the left and right ventricle volume increase, has been defined in non-beating canine heart. Ramus interventricularis anterior being firmly tethered to the myocardium undergoes about 3 times larger deformation than ramus circumflexus. In anaesthetized dogs a 30% increase in blood pressure, elicited by aortic constriction, induces an increase in diameter of coronary artery, in segment length, in blood flow and consequently in shear stress which represents a load for circumferentially running smooth muscle bundles, longitudinally running smooth muscle bundles, as well as for the endothelium. The above load lasting 4 h is already reflected by an increase in total RNA content and [14C] leucin incorporation in the left ventricle myocardium in the wall of ramus interventricularis anterior, not in ramus circumflexus. The findings fit completely with the different range of deformation of both the above coronary branches and indicates an increase in proteosynthesis not only in myocardium, but in ramus interventricularis anterior as well. An increase in ornithindecarboxylase activity in coronary wall leading to an increase in biogenic polyamines, is present in the case only, when blood pressure increase is induced by infusion of noradrenaline. PMID- 7494556 TI - Factors involved in capillary growth in the heart. AB - Growth of capillaries in the heart occurs under physiological circumstances during endurance exercise training, exposure to high altitude and/or cold, and changes in cardiac metabolism or heart rate elicited by modification of thyroid hormone levels. Capillary growth in all these conditions can be linked with increased coronary blood flow, decreased heart rate, or both. This paper brings evidence that, although increased blood flow due to long-term administration of coronary vasodilators results in capillary growth, a long-term decrease in heart rate induced by electrical bradycardial pacing in rabbits and pigs, or by chronic administration of a bradycardic drug, alinidine, in rats, stimulates capillary growth with little or no change in coronary blood flow. Decreased heart rate results in increased capillary wall tension, increased end-diastolic volume and increased force of contraction, and thus stretch of the capillary wall. This could lead to release of various growth factors possibly stored in the capillary basement membrane. Correlation was found between capillary density (CD) and the levels of low molecular endothelial cell stimulating angiogenic factor (ESAF) both in rabbit and pig hearts with CD increased by pacing. There was no relation between expression of mRNA for basic fibroblast growth factor and CD in sham operated and paced rabbit hearts. In contrast, mRNA for TGF beta was increased in paced hearts, and the possible role of this factor in the regulation of capillary growth induced by bradycardia is discussed. PMID- 7494558 TI - Oxidative stress and heart failure. AB - Various abnormalities have been implicated in the transition of hypertrophy to heart failure but the exact mechanism is still unknown. Thus heart failure subsequent to hypertrophy remains a major clinical problem. Recently, oxidative stress has been suggested to play a critical role in the pathogenesis of heart failure. Here we describe antioxidant changes as well as their significance during hypertrophy and heart failure stages. Heart hypertrophy in rats and guinea pigs, in response to pressure overload, is associated with an increase in 'antioxidant reserve' and a decrease in oxidative stress. Hypertrophied rat hearts show increased tolerance for different oxidative stress conditions such as those imposed by free radicals, hypoxia-reoxygenation and ischemia-reperfusion. On the other hand, heart failure under acute as well as chronic conditions is associated with reduced antioxidant reserve and increased oxidative stress. The latter may have a causal role as suggested by the protection seen with antioxidant treatment in acute as well as in chronic heart failure. It is becoming increasingly apparent that, anytime the available antioxidant reserve in the cell becomes inadequate, myocardial dysfunction is imminent. PMID- 7494559 TI - Structural and biochemical remodelling in catecholamine-induced cardiomyopathy: comparative and ontogenetic aspects. AB - Excessive release or administration of beta-mimetic catecholamines may induce cardiomegaly, necrotic lesions and accumulation of connective tissue in the heart of adult homoiotherms. It was examined here whether similar changes can also be observed at different stages of evolution of the cardiovascular system, i.e. in poikilotherms and in homoiotherms during embryonic life. Sensitivity of the poikilothermic hearts (carp, frog, turtle) to isoproterenol (IPRO) was significantly lower than in the homoiotherms. Necrotic lesions, if present, were localized in the inner spongious musculature which has no vascular supply but which exhibits higher activities of enzymes connected with aerobic oxidation. Moreover, the IPRO-induced decrease of the phospholipid content was also significantly more expressed in the spongious layer. IPRO treatment did not influence the total weight of the fish heart but the proportion of the outer compact layer was significantly higher. These changes were accompanied by an increase of collagen, higher water content and an increase of isomyosin with a lower ATPase activity. The response of the poikilothermic heart to IPRO-induced overload thus differs significantly from that in the homoiotherms. The administration of IPRO during embryonic life of homoiotherms (chick) induces serious cardiovascular disturbances, including cardiomegaly and cellular oedema. Necroses of myofibrils, characteristic of IPRO-induced lesions of adults, were, however, rather exceptional. IPRO did not elevate the concentration of 85Sr (as a calcium homologue) in the immature myocardium; it seems, therefore, that IPRO induced changes of the embryonic heart are not necessarily due to an intracellular calcium overload. It may be concluded that the character of catecholamine-induced cardiomyopathy is not uniform and depends strictly on the stage of cardiac development. PMID- 7494560 TI - Angiotensin converting enzyme inhibitors, left ventricular hypertrophy and fibrosis. AB - From pharmacological investigations and clinical studies, it is known that angiotensin converting enzyme (ACE) inhibitors exhibit additional local actions, which are not related to hemodynamic changes and which cannot be explained only by interference with the renin angiotensin system (RAS) by means of an inhibition of angiotensin II (ANG II) formation. Since ACE is identical to kininase II, which inactivates the nonapeptide bradykinin (BK) and related kinins, potentiation of kinins might be responsible for these additional effects of ACE inhibitors. a) In rats made hypertensive by aortic banding, the effect of ramipril in left ventricular hypertrophy (LVH) was investigated. Ramipril in the antihypertensive dose of 1 mg/kg/day for 6 weeks prevented the increase in blood pressure and the development of LVH. The low dose of ramipril (10 micrograms/kg/day for 6 weeks) had no effect on the increase in blood pressure or on plasma ACE activity but also prevented LVH after aortic banding. The antihypertrophic effect of the higher and lower doses of ramipril, as well as the antihypertensive action of the higher dose of ramipril, was abolished by coadministration of the kinin receptor antagonist icatibant. In the regression study the antihypertrophic actions of ramipril were not blocked by the kinin receptor antagonist. Chronic administration of BK had similar beneficial effects in a prevention study which were abolished by icatibant and NG-nitro-L-arginine (L-NNA). In a one year study the high and low dose of ramipril prevented LVH and fibrosis. Ramipril had an early direct effect in hypertensive rats on the mRNA expression for myocardial collagen I and III, unrelated to its blood pressure lowering effect.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7494562 TI - My experiences in Saudi Arabia. PMID- 7494561 TI - Influence of global ischemia on the sarcolemmal ATPases in the rat heart. AB - To elucidate the effect of global ischemia on the energy utilizing processes, regarding the molecular principles, the kinetic and thermodynamic properties of the sarcolemmal ATPases were investigated in the rat heart. The activation energy for hydrolysis of ATP during ischemia was higher when the reaction was catalyzed by Ca-ATPase or Mg-ATPase. For the Na,K-ATPase reaction, no changes in the activation energy were observed. With respect to the enzyme kinetics, ischemia in a time-dependent manner induced important alterations in KM and Vmax values of Na,K-ATPase, Ca-ATPase and Mg-ATPase. The Vmax value decreased significantly already after 15 min of ischemia, and it also remained low after 30, 45 and 60 min for all 3 enzymes. The significant diminution of KM values occurred later in the 30th min for Ca-ATPase, in the 45th min for Na,K-ATPase. The observed drop in KM indicates the increase in the affinity of the enzymes to substrate, suggesting thus the adaptation to ischemic conditions on the molecular level. This effect could be attributed to some conformational changes of the protein molecule in the vicinity of the ATP-binding site developing after longer duration of ischemia. PMID- 7494563 TI - Cyclophosphamide and cisplatin compared with paclitaxel and cisplatin in patients with stage III and stage IV ovarian cancer. AB - BACKGROUND: Chemotherapy combinations that include an alkylating agent and a platinum coordination complex have high response rates in women with advanced ovarian cancer. Such combinations provide long-term control of disease in few patients, however. We compared two combinations, cisplatin and cyclophosphamide and cisplatin and paclitaxel, in women with ovarian cancer. METHODS: We randomly assigned 410 women with advanced ovarian cancer and residual masses larger than 1 cm after initial surgery to receive cisplatin (75 mg per square meter of body surface area) with either cyclophosphamide (750 mg per square meter) or paclitaxel (135 mg per square meter over 24 hours). RESULTS: Three hundred eighty six women met all the eligibility criteria. Known prognostic factors were similar in the two treatment groups. Alopecia, neutropenia, fever, and allergic reactions were reported more frequently in the cisplatin-paclitaxel group. Among 216 women with measurable disease, 73 percent in the cisplatin-paclitaxel group responded to therapy, as compared with 60 percent in the cisplatin-cyclophosphamide group (P = 0.01). The frequency of surgically verified complete response was similar in the two groups. Progression-free survival was significantly longer (P < 0.001) in the cisplatin-paclitaxel group than in the cisplatin-cyclophosphamide group (median, 18 vs. 13 months). Survival was also significantly longer (P < 0.001) in the cisplatin-paclitaxel group (median, 38 vs. 24 months). CONCLUSIONS: Incorporating paclitaxel into first-line therapy improves the duration of progression-free survival and of overall survival in women with incompletely resected stage III and stage IV ovarian cancer. PMID- 7494564 TI - Blood pressure and end-stage renal disease in men. AB - BACKGROUND: End-stage renal disease in the United States creates a large burden for both individuals and society as a whole. Efforts to prevent the condition require an understanding of modifiable risk factors. METHODS: We assessed the development of end-stage renal disease through 1990 in 332,544 men, 35 to 57 years of age, who were screened between 1973 and 1975 for entry into the Multiple Risk Factor Intervention Trial (MRFIT). We used data from the national registry for treated end-stage renal disease of the Health Care Financing Administration and from records on death from renal disease from the National Death Index and the Social Security Administration. RESULTS: During an average of 16 years of follow-up, 814 subjects either died of end-stage renal disease or were treated for that condition (15.6 cases per 100,000 person-years of observation). A strong, graded relation between both systolic and diastolic blood pressure and end-stage renal disease was identified, independent of associations between the disease and age, race, income, use of medication for diabetes mellitus, history of myocardial infarction, serum cholesterol concentration, and cigarette smoking. As compared with men with an optimal level of blood pressure (systolic pressure < 120 mm Hg and diastolic pressure < 80 mm Hg), the relative risk of end-stage renal disease for those with stage 4 hypertension (systolic pressure > or = 210 mm Hg or diastolic pressure > or = 120 mm Hg) was 22.1 (P < 0.001). These relations were not due to end-stage renal disease that occurred soon after screening and, in the 12,866 screened men who entered the MRFIT study, were not changed by taking into account the base-line serum creatinine concentration and urinary protein excretion. The estimated risk of end-stage renal disease associated with elevations of systolic pressure was greater than that linked with elevations of diastolic pressure when both variables were considered together. CONCLUSIONS: Elevations of blood pressure are a strong independent risk factor for end-stage renal disease; interventions to prevent the disease need to emphasize the prevention and control of both high-normal and high blood pressure. PMID- 7494565 TI - Biliary cryptosporidiosis in HIV-infected people after the waterborne outbreak of cryptosporidiosis in Milwaukee. AB - BACKGROUND: In March 1993, the municipal water supply in Milwaukee became contaminated with cryptosporidium and a widespread outbreak of cryptosporidiosis occurred. METHODS: We retrospectively studied the relations among the clinical presentation, CD4 count, and survival in a group of 82 patients infected with the human immunodeficiency virus (HIV) in whom cryptosporidiosis developed during the outbreak. Infection was documented by the finding of cryptosporidium in stool. RESULTS: There was a sharp increase in the number of HIV-infected patients with cryptosporidiosis after the period of water contamination. Four of 24 patients with biliary symptoms (17 percent) and 30 of 58 without biliary symptoms (52 percent) were alive one year after the outbreak (P = 0.003). Twenty-one of the patients with biliary symptoms had CD4 counts < or = 50 per cubic millimeter, as compared with 36 of 57 patients without biliary symptoms (88 percent vs. 63 percent, P = 0.03). The CD4 count was not available for one patient in the latter group. Within one year, 41 of the patients with CD4 counts < or = 50 per cubic millimeter had died, as compared with 6 of those with higher counts (72 percent vs. 25 percent, P < 0.001). After adjustment for the CD4 count, independent predictors of death were older age and the presence of nausea and vomiting. CONCLUSIONS: When HIV-infected patients are exposed to cryptosporidium, those with CD4 counts < or = 50 per cubic millimeter are at increased risk for biliary symptoms and for death within one year after the infection. PMID- 7494566 TI - Brief report: late improvement in consciousness after post-traumatic vegetative state. PMID- 7494567 TI - Images in clinical medicine. Abdominal aortic aneurysm. PMID- 7494568 TI - Images in clinical medicine. Ruptured abdominal aortic aneurysm. PMID- 7494569 TI - Transcription-factor mutations and disease. PMID- 7494570 TI - Schizophrenia. PMID- 7494571 TI - DNA vaccines. PMID- 7494572 TI - Clinical problem-solving. Just in time. PMID- 7494573 TI - New therapy for ovarian cancer. PMID- 7494574 TI - Acute myocardial infarction--the value of being prepared. PMID- 7494575 TI - Drug-resistant Streptococcus pneumoniae. PMID- 7494577 TI - Oral ganciclovir for cytomegalovirus retinitis. PMID- 7494576 TI - Drug-resistant Streptococcus pneumoniae. PMID- 7494578 TI - Oral ganciclovir for cytomegalovirus retinitis. PMID- 7494579 TI - Atrial septal defect. PMID- 7494580 TI - Atrial septal defect. PMID- 7494581 TI - Atrial septal defect. PMID- 7494582 TI - Clinical problem-solving: mitral stenosis--silent? PMID- 7494583 TI - Clinical problem-solving: mitral stenosis--silent? PMID- 7494584 TI - Clinical problem-solving: mitral stenosis--silent? PMID- 7494585 TI - Neonatal diabetes mellitus. PMID- 7494586 TI - Assessment of patients' pain. PMID- 7494587 TI - Neonatal diabetes mellitus. PMID- 7494589 TI - Minorities and ESRD. Part II. Tracking causes of ESRD in the Hispanic population. PMID- 7494588 TI - Preinfarction angina as a predictor of more rapid coronary thrombolysis in patients with acute myocardial infarction. AB - BACKGROUND: When a myocardial infarction is preceded by angina, the infarct tends to be smaller than when there is no preinfarction angina. Prompt recanalization of the occluded infarct-related artery is crucial in limiting the size of the infarct. We prospectively studied the relation among preinfarction unstable angina, the speed of coronary reperfusion, and the size of the infarct in patients with acute myocardial infarction receiving thrombolytic therapy. METHODS: We compared 14 patients who had unstable angina during the week before myocardial infarction with 9 patients who had no preinfarction angina. Coronary arteriograms were obtained at base line and 15, 35, 55, and 90 minutes and 24 hours after the start of thrombolytic therapy. The size of the infarct was estimated on the basis of creatine kinase and creatine kinase MB levels, which were measured every 4 hours during the first 24 hours. RESULTS: Complete reperfusion (a flow of grade 3 according to the Thrombolysis in Myocardial Infarction classification) was achieved at 35 minutes in 64 percent of the patients with preinfarction angina but in none of those without preinfarction angina (P = 0.006); at 55 minutes in 86 percent and 38 percent, respectively (P = 0.05); and at 90 minutes in 86 percent and 50 percent, respectively (P = 0.14). The mean (+/- SD) time to reperfusion was 27 +/- 16 minutes in the group with preinfarction angina and 48 +/- 17 minutes in the group without preinfarction angina (P = 0.04); the peak creatine kinase levels were 1118 +/- 783 and 2395 +/- 1615 U per liter, respectively (P = 0.03); the peak creatine kinase MB levels were 102 +/- 67 and 251 +/- 186 U per liter, respectively (P = 0.009); and the 24 hour integrated creatine kinase MB levels were 1716 +/- 1171 and 4267 +/- 3252 U.liter-1 x 24 hours, respectively (P = 0.009). The time to reperfusion was positively correlated with the indexes of infarct size (r > or = 0.53, P < or = 0.02). CONCLUSIONS: In patients with acute myocardial infarction preceded by unstable angina, as compared with those without preinfarction angina, thrombolytic therapy resulted in more rapid reperfusion and smaller infarcts. Earlier myocardial reperfusion may thus account for the smaller infarct size in patients with preinfarction angina. PMID- 7494590 TI - Understanding Hispanic culture makes a difference at NY OPO. PMID- 7494591 TI - Hull at mid-term: a look at the next 12 months with the NKF. PMID- 7494592 TI - The value of credentialing in renal care. PMID- 7494593 TI - Reaction mixed on value of new FDA regulations for dialyzer relabelling. PMID- 7494594 TI - CA allows patient HIV tests in cases of worker exposure. PMID- 7494595 TI - A dialysis outcomes computer database. PMID- 7494596 TI - Reactive microglia in cerebral ischaemia: an early mediator of tissue damage? AB - Microglial cell activation is a rapidly occurring cellular response to cerebral ischaemia. Microglia proliferate, are recruited to the site of lesion, upregulate the expression of several surface molecules including major histocompatibility complex class I and II antigens, complement receptor and the amyloid precursor protein (APP) as well as newly expressed cytokines, e.g. interleukin-1 and transforming growth factor beta 1. The ischaemia-induced production of APP may contribute to amyloid deposition in the aged brain under conditions of hypofusion. Ultrastructurally, microglia transform into phagocytes removing necrotic neurons but still respecting the integrity of eventually surviving neurons even in the close vicinity of necrotic neurons. Microglial activation starts within a few minutes after ischaemia and thus precedes the morphologically detectable neuronal damage. It additionally involves a transient generalized response within the first 24 hours post-ischaemia even at sites without eventual neuronal cell death. In functional terms, the microglial reaction appears to be a double-edged sword in ischaemia. Activated microglia may exert a cytotoxic effector function by releasing reactive oxygen species, nitric oxide, proteinases or inflammatory cytokines. All of these cytotoxic compounds may cause bystander damage following ischaemia. Pharmacological suppression of microglial activation after ischaemia has accordingly attenuated the extent of cell death and tissue damage. However, activated microglia support tissue repair by secreting factors such as transforming growth factor beta 1 which may limit tissue damage as well as suppress astroglial scar formation. In line with ultrastructural observations microglial activation in ischaemia is a strictly controlled event. By secreting cytokines and growth factors activated microglia most likely serve seemingly opposed functions in ischaemia, i.e. maintenance as well as removal of injured neurons. Post-ischaemic pharmacological modulation of microglial intervention in the cascade of events that lead to neuronal necrosis may help to improve the structural and functional outcome following CNS ischaemia. PMID- 7494597 TI - Microglial interleukin-1 alpha expression in brain regions in Alzheimer's disease: correlation with neuritic plaque distribution. AB - Interleukin-1 alpha-immunoreactive (IL-1 alpha+) microglia are prominent components of neuritic plaques in Alzheimer's disease, and may be important in the evolution of neuritic plaques from diffuse amyloid deposits. Neuritic plaques show a characteristic distribution across cerebral regions and are absent in the cerebellum of patients with Alzheimer's disease. We used single- and dual immunohistochemical labelling to investigate the possibility that the expression of IL-1 alpha is correlated with this regional distribution of neuritic (tau 2 immunoreactive, tau 2+) plaques. In Alzheimer's disease, tau 2+ neuritic plaques occurred with increasing frequency in grey matter of frontal and occipital lobes, temporal lobe, and hippocampus. There were positive correlations between the regional patterns of distribution of activated IL-1 alpha+ microglia and tau 2+ neuritic plaques as well as between activated IL-1 alpha+ microglia and activated astrocytes. No activated IL-1 alpha+ microglia, tau 2+ neuritic plaques, or activated astrocytes were observed in cerebellum of these Alzheimer patients. These regional relationships between activated IL-1 alpha+ microglia, tau 2+ neuritic plaques, and activated astrocytes, together with the established functions of IL-1, support a causal association between the overexpression of IL 1 and the evolution of beta-amyloid deposits into neuritic plaques in Alzheimer's disease. PMID- 7494598 TI - Immortalization and characterization of rat microglial cells. AB - Microglial cell lines from rat brain were established by transfer of a temperature sensitive simian virus 40 large tumour antigen by means of a retrovirus. Four weeks after infection, colonies were generated in the presence of neomycin and granulocyte-macrophage colony stimulating factor (GM-CSF), and subsequently subcloned. Both bulk cell lines and clones proliferate actively at 33 degrees C, whereas the rate of division was significantly decreased at 39 degrees C when the large T antigen is non-functional. At 39 degrees C, these cells take on the microglial phenotype as demonstrated by immunoreactivity to ED 1 (an intracellular antigen), OX-42 (complement type 3 receptor), W3/25 (CD4 homologue), OX-6 (MHC class II antigen) and OX-18 (MHC class I antigen). These cells are capable of active phagocytosis and retain these properties for 10-15 passages. Long-term culture of these lines and clones, greater than 15 passages, displayed a gradual down-regulation of all cell surface specific antigens that were not rescued by lipopolysaccharide (LPS), interferon-gamma (gamma-IFN), GM CSF or colony-stimulating factor-1 (CSF-1). The expression of the SV-40 large T antigen was unaffected. These results demonstrate the feasibility of immortalizing short-term cell lines with the SV-40 large T antigen for their use in the characterization of microglial properties. PMID- 7494599 TI - Neurofibrillary tangles in chronic alcoholics. AB - Magnocellular neurons in the cholinergic nucleus basalis appear to be vulnerable in a variety of pathological conditions, including chronic alcoholism. While neurofibrillary degeneration of these neurons has been noted in a number of disorders characterized by dementia, the mechanism of cell death in thiamine deficient chronic alcoholics has not been identified. In the present post-mortem investigation, multiple brain regions of seven thiamine-deficient chronic alcoholics, three neurologically asymptomatic chronic alcoholics and seven non alcoholic age matched controls were screened for neurofibrillary pathology using both tau-immunohistochemistry and a modified Bielschowsky silver stain. In chronic alcoholics with thiamine deficiency, neurofibrillary pathology was found in the nucleus basalis, but not any other brain region. Neurofibrillary tangles were not seen in age-matched controls and were infrequent in alcoholics without neuropathological signs of thiamine-deficiency. Neurofibrillary tangles were most numerous in those cases with cell loss in the nucleus basalis. These findings suggest that neurodegeneration of the nucleus basalis in chronic alcoholics proceeds through the formation of neurofibrillary tangles. PMID- 7494600 TI - Distribution of tangles and threads in the cerebral cortex in progressive supranuclear palsy. AB - Recent studies have described silver- and tau-positive glia and threads in the degenerating lesions of progressive supranuclear palsy. In this study, Gallyas Braak silver impregnation and several immunohistochemical techniques were employed to examine the distribution of tangles, abnormal glia and threads in the cerebral cortex of nine cases of progressive supranuclear palsy. In addition to neurofibrillary tangles, argentophilic glia and threads were impregnated exclusively by Gallyas-Braak technique. This technique demonstrated two types of glia profiles: tightly coiled intra-cytoplasmic profiles surrounding nuclei (coiled profiles) and thorn-like profiles with radial ramifications (thorn-like profiles). Thorn-like profiles are possibly in astrocytes and were detected in the cerebral cortex, while coiled profiles are possibly in oligodendroglia and were detected both in the cerebral cortex and subcortical white matter. Topographically, many neurofibrillary tangles were constantly seen in the frontal cortex and in the pre-central gyrus. Numerous neurofibrillary tangles were detected in the entorhinal cortex of the two brains. Argentophilic glia and threads were also frequent both in the frontal cortex and the precentral gyrus; however, they were more frequent in the pre-central gyrus that in the frontal cortex in four of the eight cases examined. In two brains, argentophilic threads were distributed widely in the cerebral cortex and white matter except for the temporal cortex. In immunohistochemical studies, argentophilic glia and threads were mostly positive for Tau 2, and a small number of them were weakly positive for ubiquitin and paired helical filament protein. The immunoproperties of these abnormal glia and threads seemed to be virtually identical to those of neurofibrillary tangles.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7494601 TI - Ultrastructural abnormalities with inclusions in Onuf's nucleus in motor neuron disease (amyotrophic lateral sclerosis). AB - This study describes an ultrastructural examination focused on motor neurons in Onuf's nucleus in the spinal cord of four control patients without neurological disease (45-70 years) and six motor neuron disease (MND) patients (38-79 years; duration 8 months-19 years) who showed no somato-vesical dysfunction. Prompted by recent studies suggesting some sphincteric motor neurons may succumb to MND, this study sought to determine whether the wider population of neurons in Onuf's nucleus display ultrastructural cytopathology which is normally undetectable in histological preparations. Spinal cords were removed 3-20 h after death, and 1 mm slices of cord rapidly fixed in modified Karnovsky medium were processed for both light- and electronmicroscopy. 'Control motor neurons' had intact neuronal and nuclear membranes. Nissl bodies chiefly comprised ordered structures of alternate lamellae of rough endoplasmic reticulum and arrays of polyribosomes. The Golgi complexes consisted of multilamellated curvilinear stacks of ER. No intraneuronal filamentous or Bunina body inclusions were observed, but occasional axonal spheroids were seen in the neuropil. In MND, histological evidence of sparing in Onuf's nucleus was associated with abnormal ultrastructure of the motor neurons. Some sphincteric neurons were atrophic, whereas in the others, Nissl bodies were reduced in number, showed loss of structural organization or comprised polyribosomal aggregates. Golgi complexes had disrupted lamellated organization or consisted solely of distended ER. Intraneuronal filamentous Lewy-body or skein like inclusions and Bunina bodies were identified in Onuf's nucleus of three subjects (duration of MND 8 months-2 years). The results of the present study indicate that Onuf's nucleus is vulnerable in MND, and preservation of sphincter function with qualitative histological evidence of 'sparing' does not necessarily imply a corresponding lack of ultrastructural cytopathology in this nucleus. PMID- 7494602 TI - Single section and disector counts in evaluating neuronal loss from the substantia nigra in patients with Parkinson's disease. AB - In order to investigate the correlation between single section (SS) and disector (DS) counts for estimating the pigmented neurons in the pars compacta of the substantia nigra (SNpc), 12 patients with Parkinson's disease (PD) and seven controls were studied. The SS counts were obtained at the level of the exit of the third cranial nerve in the SNpc. The DS counts were made from the whole volume of the SNpc. There was a significant correlation between SS and DS counts (r = 0.94, P < 0.001), suggesting that the methods closely agree. Comparison of SS and DS counts in controls and PD patients showed that the pigmented neurons in PD were decreased by 75% (P < 0.001) and 55% (P < 0.001) in the SS and DS counts, respectively. This study suggests that loss of pigmented neurons from the SNpc in PD can be estimated by either method. PMID- 7494603 TI - p53 allelic imbalance in astrocytoma detected using fluorescent PCR of microsatellite repeat polymorphisms. AB - Previous reports have shown that p53 gene alteration plays an important role in tumourigenesis. Allelic loss of 17p in astrocytomas was detected in previous studies by restriction fragment length polymorphisms (RFLP). In this study we have analysed 47 cases of astrocytic tumours (26 glioblastomas [grade IV], 11 anaplastic astrocytomas [grade III], seven fibrillary astrocytomas [grade II] and three pilocytic astrocytomas [grade I]) for the presence of allelic imbalance at the p53 gene locus using intragenic markers. We used an informative method based on microsatellite polymorphisms at the p53 gene locus and fluorescent PCR. The fluorescently-labelled PCR products were then detected and analysed using an automated DNA sequencer with appropriate software. Seven of 47 (14.9%) cases were homozygous (uninformative). Five of the remaining 40 cases (12.5%) showed allelic imbalance at the p53 locus (three anaplastic astrocytomas [grade III] and two glioblastomas [grade IV]). None of the fibrillary astrocytomas (grade II) or pilocytic astrocytomas (grade I) showed allelic imbalance at the p53 locus. These results suggest that allelic imbalance at the p53 locus is not frequent and when it does occur is in high grade tumours. PMID- 7494604 TI - Apoptosis in cerebral astrocytic tumours and its relationship to expression of the bcl-2 and p53 proteins. AB - Apoptosis is an important determinant of tumour growth which can be regulated by the bcl-2 and p53 genes. This study examines the relationship between apoptosis, growth fraction (Ki-67 immunolabelling index), and accumulation of the bcl-2 and p53 proteins in a spectrum of cerebral astrocytic tumours (n = 81), including fibrillary astrocytomas (n = 16), anaplastic astrocytomas (n = 19), and glioblastomas (n = 46). Median apoptosis indices (AIs) increased across this spectrum of tumours, and a significant (P < 0.0001) correlation was demonstrated between AI and Ki-67 labelling index (LI). Immunolabelling with the bcl-2 antibody was found in 44% of fibrillary astrocytomas, 42% of anaplastic astrocytomas, and 28% of glioblastomas. It was also found in the vascular endothelial proliferation typically seen in glioblastomas, and in the giant, multinucleated cells of some glioblastomas. No clear relationship between AI and bcl-2 accumulation was evident. Immunolabelling with the p53 antibody was found in 56% of fibrillary astrocytomas, 79% of anaplastic astrocytomas, and 50% of glioblastomas. No clear relationship between AI and patterns of p53 immunolabelling was evident. Equal proportions of p53-positive tumours were bcl-2 positive and bcl-2 negative, but a small proportion of p53-negative tumours was bcl-2 positive. The correlation between AI and Ki-67 LI is in line with findings in other malignant tumours. We suggest that the regulation of apoptosis in astrocytic tumours is too complex for a clear association between AI and bcl-2 and p53 protein expression to be demonstrated. PMID- 7494605 TI - Expression and distribution of GAP-43 in human astrocytes in culture. AB - By combining mRNA analysis and immunocytochemistry, we investigated the expression of the growth associated protein 43 (GAP-43) in enriched populations of astrocytes, obtained from mixed cultures of human fetal brains. Total cellular RNA was extracted from cell pellets and reverse transcribed into cDNA; cDNA was subjected to PCR amplification using primers specific for GAP-43 and PCR products were separated through polyacrylamide gels. Double immunofluorescence staining was performed on dissociated cell cultures using antibodies to glial fibrillary acidic protein (GFAP) and to GAP-43. Results showed that both transcription and translation for GAP-43 occur in cultured astrocytes. GAP-43 immunoreacting material was detected in the cell processes and diffusely in the cytoplasm of GFAP-positive astrocytes, during early stages of maintenance in vitro. In older cultures, GAP-43 immunoreactivity persisted in a large percentage of cells, with a tendency to accumulate in perinuclear areas. These observations provide evidence that GAP-43 is not restricted to neuronal cells. The close spatial association with cytoskeletal constituents, as observed in astrocytes, suggests a role for this protein in the control of cell shape, motility and adhesion processes. PMID- 7494607 TI - Dialysis Update 1993. International workshop, 12-13 February 1993, Osaka, Japan. Proceedings. PMID- 7494608 TI - Renal osteodystrophy: pathogenesis and management. AB - Several biochemical and hormonal abnormalities associated with renal insufficiency lead to complex disorders of bone which are described by the term renal osteodystrophy. Assessment of renal osteodystrophy in its early stages is primarily biochemical since symptoms generally do not occur until osteodystrophy is advanced. Therapy should be initiated early in the course of renal insufficiency in order to prevent the development of severe skeletal abnormalities. Foremost among the multiple factors involved in the pathogenesis of hyperparathyroidism are retention of phosphorus and low levels of calcitriol. The principal therapies for the prevention and treatment of hyperparathyroidism include the use of calcium salts taken with meals, as phosphorus binders, to prevent the absorption of phosphorus from the intestine, correction of acidosis and careful use of vitamin D metabolites such as calcitriol or 1-alpha hydroxycholecalciferol. The prevalence of aluminum induced osteomalacia appears to be declining as aluminum salts have been replaced by calcium containing phosphate binders and there is increased attention to adequate water purification for dialysis. Other disorders such as adynamic bone and the accumulation of beta 2-microglobulin may require bone biopsy for accurate diagnosis and are more difficult to treat effectively. PMID- 7494606 TI - Effect of alpha-tocopherol on in vitro and in vivo metabolism of low-density lipoproteins in haemodialysis patients. AB - It has been reported that some modified low-density lipoproteins (LDLs) such as glycated LDL and malondialdehyde-rich LDL (MDA-LDL) probably exist in the circulation. The present study was undertaken to investigate the in vitro and in vivo metabolism of MDA-LDL occurring in chronic haemodialysis patients and the effects of alpha-tocopherol on these abnormalities. MDA-LDL from haemodialysis patients was degraded more rapidly by human monocyte-derived macrophages and disappeared more slowly from the circulation when compared with LDL from healthy controls. Treatment with alpha-tocopherol at doses of 600 mg/day for 2 weeks resulted in improvement of these metabolic abnormalities depending upon the degree of return to normal MDA concentrations in LDL. PMID- 7494609 TI - Pathophysiology and treatment of secondary hyperparathyroidism in patients with chronic renal failure. AB - Following four etiologies are considered as the possible reason for secondary hyperparathyroidism in the previous reports. First, a decreased serum concentration of 1,25(OH)2D3 directly stimulates PTH secretion. Second, hypocalcemia directly stimulates PTH secretion which is independent of 1,25(OH)2D3 action. Third, the presence of decreased calcemic response to PTH. Fourth, there is a strong possibility that hyperphosphatemia indirectly and/or directly may stimulate PTH secretion. The treatment of secondary hyperparathyroidism should be modified according to the stage of uremia. Excess suppression of PTH secretion could cause an adynamic bone disease. Early start of the treatment would be beneficial to prevent the bone from the development of PTH resistance. PMID- 7494610 TI - Interleukin-1 and interleukin-1 receptor antagonist production during haemodialysis: which cytokine is a surrogate marker for dialysis-related complications? AB - Studies have shown differences between host defence dysfunctions in patients with end-stage renal disease (ESRD), patients treated with continuous ambulatory peritoneal dialysis (CAPD) and those undergoing chronic haemodialysis. Immune dysfunction is well documented in haemodialysis patients, particularly after a few years of dialysis treatment. The unresolved question is whether there is a surrogate marker(s) for dialysis-related changes which can be monitored easily and which prospectively predicts deteriorating health, dialysis-related complications and/or mortality. The issue is one of finding a marker for host perturbation solely related to haemodialysis. The criteria for such a marker(s) worthy of being investigated are: (1) does it change with alterations made to the membrane or the quality of the dialysate; (2) is the marker independent of the patient's intrinsic renal disease; (3) does the marker reflect the pathophysiology of 'bioincompatibility'? In this overview, the effects of chronic haemodialysis on interleukin-1 (IL-1) and IL-1 receptor antagonist (IL-1Ra) production by peripheral blood mononuclear cells will be discussed. Recent data suggest the IL-1Ra rather than IL-1 itself is a marker for chronic monocyte activation and bioincompatibility. PMID- 7494611 TI - Effects of membrane characteristics on cytokine production by mononuclear cells in regular haemodialysis patients. AB - To determine the relationship between haemodialysis and cytokine production, the effects of solute permeability and biocompatibility of dialysis membranes on cytokine production by mononuclear cells were evaluated. Eighteen stable haemodialysis patients were divided into three groups and underwent haemodialysis under the same conditions except for the dialysis membrane used. Endotoxin in dialysate remained at concentrations of 10 pg/ml or less throughout the study. Haemodialysis was performed for a total of 6 weeks. Group A used a regenerated cellulose low-flux membrane during the first 2 weeks, a regenerated cellulose high-flux membrane during the next 2 weeks and a polymethylmethacrylate (PMMA) high-flux membrane during the last 2-week period, while Group B used the regenerated low-flux cellulose membrane first, followed by the PMMA low-flux membrane and PMMA high-flux membrane. Group C used the same membrane throughout the 6-week study period. Peripheral mononuclear cells were sampled before, 30 min after the start and upon completion of the final dialysis session and incubated for 18 h in the presence and absence of lipopolysaccharide (LPS) stimulation. Tumour necrosis factor-alpha (TNF-alpha), interleukin-1 beta (IL-1 beta) and IL-6 concentrations in the supernatant and cell lysate were determined. In all groups, cytokine production just before the final dialysis using each membrane was comparable regardless of the presence or absence of LPS stimulation. LPS stimulated TNF-alpha production decreased significantly 30 min after the start of dialysis compared to the predialysis baseline. This change was not affected by the type of membrane used.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7494612 TI - Lipid peroxidation as an indicator of biocompatibility in haemodialysis. AB - Peroxidation of cell membrane (phosphatidylcholine hydroperoxide, PCOOH) was quantitatively assessed using a chemiluminescence-HPLC system to clarify whether haemodialysis patients are damaged by oxidative stress. Patients were divided into two groups, one dialysed with conventional cellulose membranes, and the other with cellulose triacetate for 3 months, subsequently followed by a crossover study of each membrane for 3 more months. The mean value of PCOOH in haemodialysis patients was 508.5 +/- 208.7 pmol/ml (P < 0.01 vs normal controls), which showed a statistically insignificant tendency to increase during each haemodialysis session of 4 h. Two years after commencing haemodialysis, PCOOH was maintained within almost the same range, but significantly greater than normal. There was no significant tendency between an index of aortic sclerosis and plasma PCOOH. However, three patients using conventional cellulose membranes died of myocardial infarction. Plasma PCOOH increased after the commencement of haemodialysis. This tendency was more notable in patients using conventional cellulose membrane compared to the cellulose triacetate. Cellulose triacetate, which we thought to be more biocompatible, did not necessarily produce lipid peroxide. We conclude that it is very important to check production of oxygen radicals when developing new membranes. PMID- 7494613 TI - Membranes for dialysis. AB - Today, more than 30 different polymers or polymer blends are used as materials for dialysis membranes. They can be categorized following the scheme of a family tree of haemodialysis membranes. The trunk represents membranes from regenerated cellulose, major branches show either synthetically modified cellulose membranes or membranes manufactured from synthetic polymers. As the latter are standardly hydrophobic, small branches elucidate the technique on how these materials have been rendered partially or completely hydrophilic. Complications may arise, when comparing membranes only following their polymer names, such as polysulfone, polyacrylonitrile or polyamide. Due to varying polymer compositions, membranes with the same polymer names may differ in their haemocompatibility, flux properties and adsorption characteristics. Adsorption of proteins like beta 2 microglobulin, fibrinogen and coagulation factors, complement proteins, or hormones like parathormon and erythropoietin are differently adsorbed by dialysis membranes and thus adsorption contributes to the removal characteristics. Of central interest for membrane development and application is the question of how these membranes can be sterilized, as a series of patient adverse reactions has been attributed to the dialyser sterilization procedures. Apart from the cellulosic membranes Cuprophan and Hemophan, the majority of membranes cannot be sterilized by steam, as these materials degrade when exposed to above their class point temperature. Finally, future aspects of modern membrane development should not neglect the needs of patient populations with specific blood properties, such as diabetics. PMID- 7494614 TI - Pathogenetic and diagnostic aspects of dialysis-related amyloidosis. AB - Dialysis-related amyloidosis (DRA) is a major cause of morbidity in end-stage renal disease patients. While retention of the precursor protein beta 2 microglobulin (beta 2-m) forms the essential basis for DRA, pathogenetic concepts include: qualitative and quantitative alterations in beta 2-m metabolism; local and systemic inflammatory changes, partly related to different treatment modes; general predisposing factors such as age at the onset of dialysis treatment. Clinical and radiological signs, as well as synovial thickening on sonography, suggest the presence of DRA, but histomorphological demonstration of beta 2m amyloid is required for definitive proof. Scintigraphic imaging of DRA represents an additional, sensitive non-invasive diagnostic tool. Successful kidney transplantation stops the progression of DRA. PMID- 7494615 TI - Synthetic modification of PAN membrane: biocompatibility and functional characterization. AB - A disturbing interaction of PAN membranes and the bradykinin generation system particularly in the presence of angiotensin converting enzyme inhibitors has been described. A modified new membrane, SPAN (special PAN), was produced by varying the polymer components in type and composition, in particular by a reduction in Na-Methallylsulfonate. Although the SPAN membrane successfully averted the bradykinin generating ability of PAN, it was important to determine whether such a modification did not lead to a loss of the satisfactory biocompatibility profile characteristic of the parent membrane. For this purpose, we conducted the present clinical study in nine patients comparing 3 membranes; (i) a polysulphone membrane (F60S); (ii) PAN; and (iii) SPAN, to examine the clinical biocompatibility profile and performance of the new membrane. A small increase in C5a with F60S and SPAN was found which is in the range expected for highly biocompatible synthetic membranes. The three dialysers had a similar inert profile for terminal complement complex arterial values, and had similar venous values. A minimal nonsignificant decline in white cell count was observed at 15 min for all dialysers, but otherwise WBC counts were unchanged. Platelet counts were unchanged throughout treatment for the three dialysers. Arterial and venous thrombin-anti-thrombin complex values were similar for all three dialysers. F60S and SPAN dialysers had similar urea clearances.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7494616 TI - Maintaining low concentrations of plasma beta 2-microglobulin through continuous slow haemofiltration. AB - As a first step in the development of a future wearable artificial kidney, we conducted three forms of treatment with continuous haemofiltration (CHF) on a group of 10 patients in an attempt to determine what plasma values of small molecules and beta 2-microglobulin (beta 2-M) could be maintained. In the first system, one ordinary 4-h haemodialysis, followed by 7-day CHF with 51 exchange per day, was repeated. The second system consisted of CHF with 51 exchange/day and thrice weekly slow nocturnal dialysis. The third system used CHF with 101 exchange per day. The first, second and third systems were used with three, four and four patients, respectively, for at least 4 weeks each. PAN-DX and FH-66 haemofilters were used eight and three times, respectively, with continuous infusion of heparin at 500-700 units/h. In the first system, the pre-CHF plasma beta 2-M was 45.5 +/- 3.1 mg/l, against a much lower 18.8 +/- 1.0 mg/l 4 weeks later. With the second system, this decrease was from 43.5 +/- 5.7 mg/l to 18.1 +/- 1.1 mg/l with 4 weeks treatment, and from 44.7 +/- 3.4 mg/l to 18.1 +/- 1.4 mg/l with 3 weeks treatment. Pre-haemodialysis treatment levels of urea nitrogen, plasma creatinine and uric acid in the first system were greater than ordinary pre-haemodialysis levels. On the second system, these were kept at -54 +/- 8.3%, 52.2 +/- 6.7% and -47.6 +/- 4.4% of the ordinary pre-haemodialysis levels, respectively.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7494617 TI - The influence of dialysis time and dialyser reuse on survival. PMID- 7494618 TI - Diagnostic radionuclide imaging of dialysis-related amyloidosis. PMID- 7494619 TI - Risk assessment of essential trace elements: new approaches to setting recommended dietary allowances and safety limits. AB - By definition, every essential trace element must have a range of intakes safe from toxicity but adequate enough to meet nutrition requirements. That range is part of the total dose-response curve and its lower and upper limits are delineated on the basis of nutrition and toxicology data, respectively. Close coordination of activities to set these limits is necessary to avoid recommendations that are either impractical (narrow zones of safe and adequate intakes) or contradictory (overlapping limits, i.e., no zones of safe and adequate intakes). PMID- 7494620 TI - Complex carbohydrates: the science and the label. AB - This conference resulted in a foundation upon which consensus will be built in the future. Further, it provided a forum for the articulation of the critical need for carbohydrates in human nutrition. We have come a long way in the past 25 years in the food and nutrition sciences and this conference well illustrated that point. At the White House Conference on Nutrition in 1969, macronutrients were never even mentioned as being necessary to nutrition; only micronutrients were emphasized. In 1977 the term complex carbohydrate was used without definition in the Dietary Goals for the United States. This conference encapsulated the tremendous strides that have been made concerning the chemical, nutritional, biologic, and physiologic importance of carbohydrates in health and disease. It is fascinating to review the recommendations concerning complex carbohydrate from 1977 on. Recommendations were made by the Senate Select Committee in 1977; the Dietary Guidelines of 1980, 1985, and 1990; the Surgeon General's report of 1979 and 1988; the 10th edition of Recommended Dietary Allowances; the 1989 National Academy of Sciences Diet and Health: Implications for Reducing Chronic Disease Risk; the 1991 FDA proposal for labeling; and the 1992 Food Guide Pyramid. Indeed, as Dr. Joanne Slavin pointed out, "there is a wide range of consensus that increased complex carbohydrate is consistent with good health," and certainly as Dr. Gary Henderson said, "the term complex carbohydrate has equity with consumers." With this level of apparent clarity it is difficult at first to identify a problem for the food label. After all, if consumer health would benefit, why not simply do it? However, a problem does exist in that carbohydrates may be classified chemically in groups that are difficult to analyze and to define as to their physiologic function in the body. Indeed, complex carbohydrates defined by chain length may have molecular structures less complex than those of smaller units and may vary in solubility and digestibility. A continuum does exist in each of the chemical, analytical, and physiologic characteristics of carbohydrates. Scientists do not like a continuum and regulators probably like it even less, but such a continuum holds great promise for future consensus. This continuum provided the thoughtful alternatives presented above by Drs. Jonathan DeVries, Dennis Gordon, and Alison Stephen. Dr. DeVries suggested a scheme of complex carbohydrates, sugars, and sugar alochols, while Dr. Gordon suggested complex carbohydrates, dietary fiber, and simple carbohydrates, and Dr. Stephen suggested starch, sugar, and nonstarch polysaccharides.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 7494621 TI - alpha-Linolenic acid: a preventive in secondary coronary events? AB - In a 5-year study of survivors of a first myocardial infarction, the effect of an alpha-linolenic acid-enriched diet (experimental) was compared with the prudent diet of the American Heart Association (control) in prevention of secondary coronary events. After a mean follow-up of 27 months, there were 5 nonfatal myocardial infarctions and 3 cardiac deaths in the experimental group versus 17 nonfatal myocardial infarctions and 16 cardiac deaths in the control group. Overall mortality was 20 in the control and 8 in the experimental group. An alpha linolenic acid-enriched diet appears to be effective in secondary prevention of coronary events, but a follow-up trial with a larger sample size is suggested. PMID- 7494623 TI - WHO and FAO joint consultation: fats and oils in human nutrition. PMID- 7494622 TI - Hyperhomocysteinemia as a risk factor for cardiovascular disease in patients undergoing hemodialysis. AB - Hyperhomocysteinemia exists among patients both in end-stage renal disease and on dialysis and may represent an additional risk factor for increased cardiovascular disease. Supplementation with folic acid may reduce, but not correct, hyperhomocysteinemia. Evidence that lowering blood homocysteine will lessen cardiovascular risk is being sought. PMID- 7494624 TI - Stress and the female doctor. PMID- 7494626 TI - Information technology: a layman's guide to IT. PMID- 7494625 TI - A system for life. PMID- 7494627 TI - E-mail = mc2. PMID- 7494628 TI - Confessions of an Internet addict. PMID- 7494629 TI - Apologise? Not likely! PMID- 7494630 TI - Counselling and the role of OH. PMID- 7494631 TI - Writing for publication. PMID- 7494632 TI - Taking the fear out of presentations. PMID- 7494633 TI - New contracts for old. PMID- 7494634 TI - The search for missing links between self-replicating nucleic acids and the RNA world. AB - The notion that modern metabolism was derived from a complex 'RNA world' in which most reactions were catalyzed by ribozymes has received wide acceptance. However, the evolutionary links between the first self-replicating systems and ribozymes as complex as, say, the Group I self-splicing intron or the HDV ribozyme, have remained elusive. While prebiotic chemists have succeeded in synthesizing short oligonucleotides, it is not immediately obvious how these could have replicated and evolved to the point where they could assume complex shapes and catalytic functions. Nonetheless, recent experiments from a variety of disciplines suggest a plausible pathway from prebiotic chemistry to complex metabolism, and this review is intended as a hypothetical roadmap for the origin and subsequent evolution of life. PMID- 7494635 TI - The phylogeny of tRNAs seems to confirm the predictions of the coevolution theory of the origin of the genetic code. AB - An extensive analysis of the evolutionary relationships existing between transfer RNAs, performed using parsimony algorithms, is presented. After building up an estimate of the tRNA ancestral sequences, these sequences are then compared using certain methods. The results seem to suggest that the coevolution hypothesis (Wong, J.T., 1975, Proc. Natl. Acad. Sci. USA 72, 1909-1912) that sees the genetic code as a map of the biosynthetic relationships between amino acids is further supported by these results, as compared to the hypotheses that see the physicochemical properties of amino acids as the main adaptative theme that led to the structuring of the genetic code. PMID- 7494636 TI - Two types of aminoacyl-tRNA synthetases could be originally encoded by complementary strands of the same nucleic acid. AB - The lack of even a marginal similarity between the two aminoacyl-tRNA synthetase (aaRS) classes suggests their independent origins (Eriani et al., 1990; Nagel and Doolittle, 1991). Yet, this independence is a puzzle inconsistent with the common origin of transfer RNAs, the coevolutionary theory of the genetic code (Wong, 1975, 1981) and other associated data and ideas. We present here the results of antiparallel 'class I versus class II' comparisons of aaRSs within their signature sequences. The two main HIGH- and KMSKS-containing motifs of class I appeared to be complementary to the class II motifs 2 and 1, respectively. The above sequence complementarity along with the mirror-image between crystal structures of complexes formed by the opposite aaRSs and their cognate tRNAs (Ruff et al., 1991), and the generally mirror ('head-to-tail') mapping of the basic functional sites in the sequences of aaRSs from the opposite two classes led us to conclude that these two synthetases emerged synchronously as complementary strands of the same primordial nucleic acid. This conclusion, combined with the hypothesis of tRNA concerted origin (Rodin et al., 1993a,b), may explain many intriguing features of aaRSs and favor the elucidation of the origin of the genetic code. PMID- 7494637 TI - Rural cancer care think tank uncovers significant issues/concerns. PMID- 7494638 TI - Is nursing a good career choice for the 21st century? PMID- 7494639 TI - New JCAHO standard addresses conscientious objection. PMID- 7494640 TI - Immunoprophylactic potential of a 120 kDa Brugia malayi adult antigen fraction, BmA-2, in lymphatic filariasis. AB - A 120 kDa antigen containing SDS-PAGE fraction BmA-2 isolated from Brugia malayi adult parasite was highly reactive with normal sera from filarial endemic area. BmA-2 was analysed for its prophylactic potential in in vitro and in vivo. Sera collected from BmA-2 immunized jirds induced a significant level (80 to 90%) of protection against infective larvae and microfilariae in in vitro ADCC assay as well as in in situ micropore chamber implantation studies. Mastomys natalensis immunized with BmA-2 showed a significant level of protective response against circulating microfilariae by clearing 90% of them from circulation by fifth day after challenge infection. Immunization of jirds with BmA-2 resulted in an enhanced level of antibody response against BmA-2 and 88% reduction in the development of the parasites to the adult stage. Passive transfer of immunesera from jirds immunized with BmA-2 to naive jirds resulted in 71% reduction in adult worm recovery as observed 90 days after challenge infection with B. malayi. On the other hand the passive transfer of non-adherent spleen cells from immune jirds did not show any significant effect on the development of parasite. Administration of jirds anti BmA-2 serum to microfilaraemic jirds showed a temporary decrease in microfilarial count which was increased to pretherapeutic level within 100 days and there was no effect on the adult worms. This implies that the immune protective effect of BmA-2 is mainly antibody dependent and active immunization with BmA-2 is effective against filarial infection. PMID- 7494641 TI - Further characterization of the 38 kDa antigen from Echinococcus granulosus (hydatid disease) cyst fluid: evidence for antigenic heterogeneity and reactivity with anti-P1 antibodies. AB - A panel of 5 IgM and 6 IgG1 monoclonal antibodies (MoAbs) was produced against a band, eluted from a reducing SDS-PAGE gel, containing the 38 kDa subunit of antigen 5 (Ag 5) from Echinococcus granulosus cyst fluid; seven of the MoAbs were shown subsequently to bind epitopes on Ag 5 but none recognized phosphorylcholine or periodate-sensitive carbohydrate epitopes. Differences in the fine specificity of the MoAbs were apparent and, upon reduction, heterogeneity in 38 kDa components from hydatid cyst fluids of different intermediate host origin was revealed by peptide fingerprinting and immunoblotting using the MoAbs. One of the IgG1 MoAbs (ED9) was able to distinguish a reduced 38 kDa molecule in cyst fluids from two distinct genotypes--the horse/dog and sheep/dog strains--of E. granulosus and this may have implications for hydatid serology, immunoepidemiology and strain typing. Furthermore, epitopes on this 38 kDa component or on a different molecule with the same or similar M(r) are reactive with anti-P1 blood group antigen antibodies and this could result in false positive reactions where sera from P2 patients with suspected hydatid disease are tested by immunoblot or immunoprecipitation analysis. PMID- 7494642 TI - Comparison of antibody isotype responses to Schistosoma mansoni antigens by infected and putative resistant individuals living in an endemic area. AB - The isotypic patterns of antibodies against Schistosoma mansoni antigenic preparations from eggs (SEA), adult worms (SWAP) and cercariae (CERC) have been studied in sera from two groups of individuals living in an area endemic for S. mansoni. One of the groups was comprised of individuals diagnosed as having S. mansoni infections based on their patency, i.e. those passing eggs in their faeces (patent infections, PI). The other group has been consider 'putatively resistant' due to their residence in an endemic area, their documented exposure to positive transmission sites, and their repeated negativity upon stool examinations (endemic normals, EN). There are strong specific responses of IgG1, IgG4 and IgM, particularly to SEA and CERC, by both groups. The reactivities of all isotypes were lower to SWAP. The responses of IgG4, IgM and IgE anti-CERC in EN and PI are higher than those found in normal individuals from outside endemic areas. In general, EN individuals express a relative higher level of anti-STEG IgE as compared to IgG4. On the other hand the pool of sera from PI showed the opposite pattern of higher IgG4 as compared to IgE. Several correlations are seen between isotypic responses to SEA, SWAP and CERC based on comparisons to the anti SWAP IgE responses of the individuals in the two groups. These comparisons indicate the presence of distinct immunologic differences between individuals in the PI and the EN groups. PMID- 7494643 TI - Plasmodium vivax: detection of blood parasites using fluorochrome labelled monoclonal antibodies. AB - A monoclonal antibody (MoAb) 2C6111 specific for Plasmodium vivax erythrocytic stages was shown to detect parasitized erythrocytes in blood samples collected in the field. This MoAb binds to the mature trophozoite, schizont and gametes of P. vivax and upon examination of 43 wild isolates no evidence of polymorphism was found. To search for P. vivax parasites in human blood a MoAb immunofluorescent test (MoAb-IFT) was developed. The assay is based on the ability of fluorescein isothiocyanate labelled MoAb 2C6111 to combine with parasitized erythrocytes on thin blood smears. A preliminary field trial was carried out in Venezuela to determine the usefulness of MoAb-IFT for the specific diagnosis of P. vivax malaria. Blood samples collected from malarious and non-malarious individuals were examined both by standard microscopy of Giemsa stained thick blood smears (G TS) and MoAb-IFT. The latter was specific and gave a 100% correlation with G-TS. Sensitivity was close to that usually achieved with Giemsa stained blood films. PMID- 7494644 TI - Immunoepidemiology of human necatoriasis: correlations between antibody responses and parasite burdens. AB - In this paper we describe the relationship between antibodies to Necator americanus stage-specific antigens and parasite burden in an endemically-infected population in Papua New Guinea. Using an age-structured analysis, we show that the correlation coefficient between levels of IgG against adult worm excretory secretory (ES) antigen and parasite burden declined significantly with host age from positive in younger hosts to significantly negative in older hosts. A trend towards similar patterns was present for anti-larval IgG both pretreatment and after reinfection, and for anti-ES IgM and anti-ES IgE pretreatment. These patterns are consistent with a role for these isotypes in a protective immune response, although parasite-induced immunosuppression may provide an alternate explanation. This is another demonstration of possibly protective responses to N. americanus infection. PMID- 7494645 TI - Humoral immune response of children with chronic schistosomiasis. Isotype recognition of adult worm antigens. AB - In areas of low transmission of schistosomiasis, the evaluation of the success of control depends on reliable diagnostic tests. Under such conditions, some of the serological tests better estimate the real prevalence of this parasitosis than the classical stool examinations. On the search of highly sensitive and specific antigenic fractions for use in serological tests, an immunoblot technique with a luminescent substrate was used in order to evaluate, under dissociating and reducing conditions, the Schistosoma mansoni adult worm antigen (AWA). The sera of 30 infected Venezuelan children were assayed for specific recognition of AWA by IgG, IgM, IgE, IgA, and the four IgG subclasses. Eight highly specific polypeptide molecules from the parasite of 118, 114, 105, 74, 71, 45, 36, and 30 kDa were recognized by total IgG. Additionally, IgG1 and IgG2 recognized a molecule of 100 kDa and IgM one of 77 kDa. The present data suggests that certain molecules from the adult worm, specially the 36 kDa, might be relevant in the specific immunodiagnosis of this parasitic disease. The fact that the children antibodies were able to recognize the primary structure of these antigens, will allow us to chemically synthesize the relevant linear epitopes. PMID- 7494647 TI - Acute hyperoxic injury attenuates the relaxing effects of "loop" diuretics and salbutamol on large airways of newborn guinea pigs. AB - We have previously found an age-dependent relaxing effect of furosemide in normal fetal, newborn, and adult guinea pig airways with fetal trachea exhibiting the greatest relaxation and adult tissue the least. This study was designed to expand upon this finding by determining if in vivo hyperoxic exposure would influence in vitro airway relaxation mediated by the loop diuretics, furosemide and ethacrynic acid, and the beta 2-adrenoceptor agonist, salbutamol. Newborn guinea pigs were raised in > 95% FiO2 until ill; controls in room air. Isometric relaxation to 3 x 10(-5) M furosemide, 3 x 10(-6) M ethacrynic acid, or 10(-8)-10(-6) M salbutamol was recorded in 3 x 10(-6) M histamine-constricted airway rings. Ethacrynic acid, like furosemide, relaxed newborn guinea pig airways. Hyperoxia did not alter the contractile effect of 3 x 10(-6) M histamine but did significantly decrease the relaxing effect of furosemide, ethacrynic acid, and salbutamol. Loop diuretic mediated airway relaxation was accentuated in HEPES buffer when compared with Krebs, whereas salbutamol-mediated relaxation was unaffected. These results suggest that hyperoxia nonspecifically decreases airway responsiveness to the relaxing agents studied. PMID- 7494646 TI - HLA-derived peptides as novel immunosuppressives. AB - Peptides corresponding to linear sequences of HLA molecules have been synthesized and tested for immunomodulatory activity in in vitro assays using human T lymphocytes. Sequences from different parts of the HLA molecules have different effects. Peptides corresponding to residues 75-84 of an HLA class I supratypic specificity of limited heterogeneity (HLA-Bw4) had profound inhibitory effects in a variety of in vitro assays of human T lymphocyte function. Furthermore, a 2-wk course of human HLA sequences and cyclosporine therapy induced enduring immunologic tolerance in a rat model of heterotopic heart transplantation. These studies prompted clinical trials which are currently in progress. The peptides appear to induce T cell anergy by causing a prolonged intracellular calcium flux and interrupting normal signal transduction pathways. Furthermore, these peptides bind to members of the heat-shock protein 70 family, implicating these ubiquitous proteins in the immunomodulatory pathway. Such peptides may be normal physiologic mediators. In any case, they represent potential new immunotherapeutics for a variety of immune-mediated diseases. PMID- 7494648 TI - Hyperoxic injury of immature guinea pig lung is mediated via hydroxyl radicals. AB - Support of preterm infants with ventilation and oxygen therapy frequently leads to the development of chronic lung disease. Oxidative stress, through the generation of excess oxygen free radicals, is thought to play a major role in this condition. At present the radical species responsible for oxidative lung injury is not known, and effective antioxidant based therapies are not available. The purpose of this study was to determine whether hydroxyl radicals, potent reactive oxygen species, are involved in chronic oxidative lung injury. To obtain this information we developed a animal model of chronic lung injury using the preterm guinea pig and analyzed lung tissue from these pups for o-tyrosine, a specific marker of hydroxyl radical attack. In normoxia control pups the pulmonary content of o-tyrosine was low during the first 4 wk of life (range 0.11 0.12% tyrosine). Pups maintained in 85% oxygen were found to have increasing lung o-tyrosine over this period (d 7, 0.51%; d 14, 0.8%; d 21, 1.28%; d 28, 1.45% tyrosine). From d 21, the nonenzymatic glycosylation end product, N-epsilon carboxymethyllysine was also present in significantly increased amounts in hyperoxic-exposed pups. These results implicate hydroxyl radicals as a significant oxidizing species in hyperoxic lung injury and provide a basis for understanding collagen deposition in the neonatal lung. PMID- 7494650 TI - Submental and diaphragmatic muscle activity during and at resolution of mixed and obstructive apneas and cardiorespiratory arousal in preterm infants. AB - Pathomechanisms involved in obstructive apneas remain obscure. Apnea arousal failure has been proposed as a cause for sudden death during sleep. The present study hypothesizes an interdependency between upper airway dilating submental muscle electromyogram (EMG) activity (EMGsub), diaphragmatic muscle activity (EMGdia), incidence of bradycardia, and transcutaneous measured PO2 (tcpO2) upon termination of apnea. Polygraphic recordings, including surface EMG (EMGsub, EMGdia), EEG, ECG, and transcutaneous PO2/PCO2 (tcpO2/tcpCO2) were performed on 10 preterm infants at 36, 40, 44, and 52 wk of conceptional age. EMGsub increased initially, then decreased in 28 of 33 non-rapid eye movement (N-REM) sleep apneas (REM: 35 of 69 events). This correlated with a decrease of tcpO2 during N-REM sleep (p < 0.05). A parallel decrease of EMGsub and EMGdia was correlated with the occurrence of bradycardia (REM and N-REM: p < 0.01). Concomitant termination of apnea and bradycardia (n = 22), occurred in the presence of a phasic, simultaneous activation of EMGsub and EMGdia in 64% of REM sleep and in 79% of N REM sleep-related event, was characterized by a deep inspiration preceded by a short expiration, and correlated with the extent of tcpO2-decline during REM sleep apneas (p < 0.05). In one apnea with bradycardia that progressed to asystolia, this mechanism was missing, but was evoked by a slight tactile stimulation, where-upon cardiorespiratory functions were immediately reestablished whereas N-REM sleep continued uninterrupted. Our data demonstrate an interdependency between changes of EMGsub and EMGdia activity, tcpO2 decline, and occurrence of bradycardia. A "cardiorespiratory arousal" terminated apneas and bradycardia without a change in sleep phase. PMID- 7494649 TI - Premature rats treated with propylthiouracil show enhanced pulmonary antioxidant enzyme gene expression and improved survival during prolonged exposure to hyperoxia. AB - In full-term newborn rats, propylthiouracil (PTU) treatment has been previously shown to decrease susceptibility to O2-induced lung damage and improve survival during hyperoxic exposure. However, no differences were found in lung antioxidant enzyme (AOE) activity responses to hyperoxia compared with O2-exposed untreated (control) term rats. To further explore possible pulmonary protective effects of PTU treatment in prematurely delivered animals, we administered PTU (0.015%) in drinking water to timed-pregnant rats for the final 10 d of gestation prior to delivery 1 d before term, and during lactation; control pregnant/nursing rats received untreated water. Both groups of 21-d premature rat pups were randomized to either > 95% O2 or room air exposure after birth for up to 14 d. The left lungs of 7-d exposure pups were used to quantitate the concentrations of AOE mRNA by solution hybridization; the right lungs of the same pups were assayed for AOE activities. PTU treatment resulted in survival rates of O2-exposed preterm rat pups that were consistently higher at all time periods in hyperoxia including 7 d [PTU, 67 of 82 (82%) versus control pups, 58 of 113 (51%); p < 0.001] and 14 d [PTU, 31 of 39 (79%) versus control, 15 of 66 (23%); p < 0.001]. Further evidence of increased tolerance to > 95% O2 in PTU pups included a significant decrease in the incidence of microscopic intraalveolar edema and a significant increase in lung tissue surfactant-related phospholipids compared with O2-exposed control pups.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7494651 TI - Reduction in cerebral ischemic injury in the newborn rat by potentiation of endogenous adenosine. AB - Because of ontogenic influences on the pathophysiologic mechanisms of brain injury in the perinatal brain, and in particular, the incomplete development of adenosine receptor systems, we investigated the potential for adenosine to provide cerebro-protection in a well established newborn rat model of hypoxia ischemia. Fifteen litters of postnatal d 7 animals were subjected to unilateral carotid ligation and exposure to hypoxia (8% oxygen) for 3 h. Immediately after hypoxia-ischemia, animals received either the adenosine deaminase inhibitor deoxycoformycin (DCF; 2.5 mg/kg intraperitoneally) or the adenosine uptake inhibitor propentofylline (PPF; 10 mg/kg intraperitoneally); paired littermates received an equivalent volume of normal saline. On postnatal d 14, injury or protection was assessed by differences in hemispheric weights, morphometric determinations of infarct area, and histopathologic analyses. DCF resulted in a 34% (p = 0.02) and 31% (p = 0.03) reduction in hemispheric weight disparities and infarct area, respectively; for PPF, these reductions were 46% (p = 0.03) and 32% (p = 0.04), respectively. Light microscopic examinations of striatum, thalamus, hippocampus, and cortex revealed that both drugs significantly improved histologic scores as well. Measurements in six separate litters indicated that neither drug significantly reduced core body temperature for at least 6 h postadministration. These findings indicate that potentiation of endogenous adenosine levels in the perinatal brain can significantly ameliorate brain injury. Each of these treatment strategies was effective even when administered after the hypoxic-ischemic insult. Thus, further investigations of adenosinergic therapies are warranted in this and other perinatal models of cerebral ischemia to elucidate in detail their potential for clinical application. PMID- 7494652 TI - The effect of long term caffeine treatment on hypoxic-ischemic brain damage in the neonate. AB - There is considerable concern over the widespread use of caffeine during and after pregnancy. We have therefore examined the effect of perinatal caffeine use on the vulnerability of the immature brain to hypoxic ischemia (HI). Rat pups were exposed to caffeine during the first 7 d after birth by addition of a low or a high dose (0.3 or 0.8 g/L) of caffeine to the drinking water of their dams. At 7 d the pups were exposed to unilateral carotid occlusion+exposure to 7.70% oxygen for 100 min. The extent of HI brain damage was evaluated 2 wk after the insult. The effects of caffeine on A1 and A2a receptors, A1 mRNA and A2a mRNA, were examined by receptor autoradiography and in situ hybridization. Caffeine, theobromine, theophylline, and paraxanthine were analyzed in plasma of separate animals. Exposure to caffeine reduced HI brain damage from 40.3 +/- 3.2% in controls to 29.8 +/- 4.0% (p < 0.05) in low dose and 33.7 +/- 3.9% (NS) in the high dose group. The A1 receptor density measured as [3H]-1,3-dipropyl-8 cyclopentyl xanthine ([3H]-DPCPX) binding was not significantly affected after low dose caffeine but increased in the brain of rat pups in the high dose group. The A2a receptor density measured as [3H]-2[p-(2-carbonylethyl)-phenethylamino] 5'-N- ethylcarboxamidoadenosine ([3H]-CGS 21680) binding and the expression of A1 mRNA and A2a mRNA were not altered by caffeine treatment. In conclusion, low dose caffeine exposure (plasma levels corresponding to umbilical cord plasma in newborns of coffee-consuming mothers) reduced HI brain damage by 30% in 7-d-old rats.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7494653 TI - Measurement of cerebral venous oxyhemoglobin saturation in children by near infrared spectroscopy and partial jugular venous occlusion. AB - Changes in cerebral venous oxyhemoglobin saturation reflect changes in the balance between cerebral oxygen delivery and cerebral oxygen consumption. Invasive monitoring of cerebral venous saturation (CSVO2) has provided useful information in the management of critically ill adults at risk of cerebral hypoxia. This study describes the development and validation of a non-invasive method of measuring CSVO2 suitable for use in sick neonates using near-infrared spectroscopy (NIRS) and partial jugular venous occlusion. This technique was validated by comparison with an invasive measurement of CSVO2, co-oximetry of jugular bulb blood obtained during cardiac catheterization. Agreement between the two methods was assessed using the method of J. M. Bland and D. G. Altman. Fifteen children were studied, aged 3 mo to 14 y (median 2 y). CSVO2 by co oximetry ranged from 36 to 80% (median 60%). The mean difference (Co-Oximeter - NIRS) was 1.5%. Limits of agreement were -12.8 to 15.9%. Three different methods of analyzing the NIRS signal were compared. The best agreement was obtained when the changes occurring during the first 5 s of partial jugular venous occlusion were studied. Greatest accuracy was seen in those subjects with least movement artifact, and we believe this technique will be reliable in sick neonates. PMID- 7494654 TI - Diagnosis of inborn errors of metabolism from blood spots by acylcarnitines and amino acids profiling using automated electrospray tandem mass spectrometry. AB - Acylcarnitine profiling from blood or plasma samples by electrospray tandem mass spectrometry (ESI-MS/MS) has been recognized recently as a useful tool in the biochemical diagnosis of propionic acidemia, methylmalonic acidemia together with short-chain and medium-chain acyl-CoA dehydrogenase deficiencies. In the current study, we have investigated the diagnostic capabilities of ESI-MS/MS in other types of organic acidemias and amino acid catabolism disorders. Using multiple scanning functions, we examined the potential for the simultaneous profiling of both acylcarnitines and amino acids, in each of the samples. Our method was found to be specific and accurate; allowing quantification of acylcarnitines and amino acids well below, and significantly above, published normal levels. Complete automation of sample introduction has been achieved, allowing the analysis of up to 200 samples in one injection sequence, at a rate of one sample every 3 min, with excellent separation between successive injections. In our hands, this method permits screening for 20 organic acid and amino acid disorders, using a single sample injection. In our laboratory, more than 2000 blood samples have been analyzed, and 52 new cases were diagnosed by this method. We also confirmed the diagnosis of another 75 previously known cases. PMID- 7494655 TI - Sucrose reduces pain reaction to heel lancing in preterm infants: a placebo controlled, randomized and masked study. AB - In term infants sucrose given by mouth has been reported to reduce duration of crying after a heel prick. This study was designed primarily to investigate the effect of sucrose administered orally immediately before heel lancing on the nociceptive reaction in preterm infants as assessed by change in heart rate and duration of crying. A secondary objective was to document changes in cerebral blood volume during acute pain. We used a randomized, masked, placebo-controlled, crossover trial in a neonatal intermediate care unit in a level 3 perinatal center. The patients studied were 16 preterm infants; birth weight, 900-1900 g; gestational wk, 27-34; corrected postmenstrual age at time of investigation, 33 36 wk. Each infant was assessed twice receiving 2 mL of sucrose 50% or 2 mL of distilled water in random order immediately before heel lance. Heart rate, thoracic movements, and transcutaneous blood gases were monitored continuously. Crying during the procedure was documented by a video-camera. A change in cerebral blood volume was assessed by near-infrared spectroscopy. We found the heart increased by a mean of 35 beats/min (bpm) after sucrose and 51 bpm after placebo (median difference 16 bpm, interquartile range 1-30 bpm, p = 0.005). Infants cried 67% of time after sucrose and 88% after placebo (median difference 10%, interquartile range 3-33%, p = 0.002). Cerebral blood volume decreased in 5 of 14 infants after sucrose and in 6 of 14 infants after placebo (difference not significant). PMID- 7494656 TI - Soluble L-selectin (sCD62L) umbilical cord plasma levels increase with gestational age. AB - L-Selectin (CD62L) is a leukocyte surface membrane glycoprotein involved in extravasation and homotypic aggregation which is rapidly cleaved off after cellular activation. From culture supernatants and body fluids, soluble L selectin (sCD62L) has been recovered with its functional activity retained. We devised a sensitive enzyme-linked immunoassay for quantitation of sCD62L which was used to measure sCD62L in umbilical cord plasma of 255 human newborns with a gestational age (GA) of 23-43 wk (median 38 wk). sCD62L levels ranged from 1.14 13.8 pmol/mL (median 7.2 pmol/mL) and showed strong correlations with GA (r = 0.71, p < 0.001), birth weight (r = 0.66, p < 0.001), and absolute neutrophil cell counts (ANC) (r = 0.62, p < 0.001) obtained from a peripheral vein within the first 6 h of life (n = 153), whereas there was a weak inverse correlation with absolute normoblast counts (r = -0.27, p < 0.001). In multiple regression analysis, only GA and ANC retained a significant association with sCD62L levels (p < 0.001). Decreased sCD62L levels were found to be associated with multiple gestation (4.8 +/- 2.4 pmol/mL versus 7.7 +/- 2.3 pmol/mL, p < 0.05) also when considering GA and ANC as covariates. In contrast, increased sCD62L levels in infants born from meconium-stained amniotic fluid, and decreased levels in newborns with acute bacterial infection could be fully attributed to differences in GA and ANC.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7494657 TI - Hemodynamic changes in HH stage 34 chick embryos after treatment with all-trans retinoic acid. AB - To obtain insight into the hemodynamics of abnormal cardiac development, a chick embryo model was recently developed in which a spectrum of double outlet right ventricle was induced with all-trans-retinoic acid. In Hamburger and Hamilton (HH) stage 34 white Leghorn chick embryos, we simultaneously measured dorsal aortic flow velocities with a 20 MHz pulsed Doppler velocity meter and vitelline artery blood pressures with a servonull system. These measurements were performed in embryos treated at HH stage 15 with 1 microgram of all-trans-retinoic acid (n = 47), or with the solvent DMSO (n = 15), and in control embryos (n = 21). After the wave form recordings were collected, all embryos were examined histologically. Embryos treated with all-trans-retinoic acid showed in 15 cases hearts with a rightward positioned aorta with an additional subaortic ventricular septal defect and 32 cases without septation abnormalities of the heart. The hemodynamic data were correlated with the morphology. Statistical comparison was performed between control and experimental values. There was no significant discrepancy in hemodynamics of sham-operated and control embryos. Heart rate, peak systolic and mean velocities, peak systolic and mean blood flows, and peak acceleration and stroke volume were reduced in embryos treated with all-trans retinoic acid (p < 0.01). Furthermore, in the presence of a subaortic ventricular septal defect the diameter of the dorsal aorta was reduced. Pressure readings were not statistically significant. Our findings suggest that the hemodynamic changes are the result of a decrease in cardiac contraction force. PMID- 7494658 TI - Bile pigment composition and bilirubin esterification in the developing chick. AB - In vivo and in vitro bile pigment metabolism was studied in embryos, chicks, and mature White Leghorn hens. Biliverdin and bilirubin pigments were present in bile of embryos from the earliest stage examined (14 d). Bilirubin accounted for 2.5 11.5% of total bile pigments, with the higher percentages in the early embryo. Biliverdin was exclusively in unesterified form. In contrast, bilirubin was almost entirely (> 98%) in esterified form. Glucosides consistently predominated over glucuronides and xylosides (6:3:1). In serum, bilirubin and biliverdin were undetectable at all embryonic stages and after hatching. Bilirubin UDP glycosyltransferase activities with UDP-glucuronic acid, UDP-glucose, and UDP xylose were detectable in chick embryo liver and averaged 67, 72, and 102%, respectively, of the corresponding adult mean values, without significant change throughout development. We conclude that multiple bilirubin esterification systems mature early in the avian embryo. This is in marked contrast to the development of bilirubin metabolism in the mammalian fetus in which detoxication of this potentially toxic pigment is achieved by placental clearance, and bilirubin esterification matures only after delivery. These contrasting developmental patterns are consistent with an adaptive response to the different requirements in the avian embryo and mammalian fetus for self-protective detoxification of unconjugated bilirubin. PMID- 7494659 TI - Regulation of amiloride-sensitive Na+ transport in immature rat distal colon by aldosterone. AB - The effects of dietary changes and plasma aldosterone levels on channel-mediated electrogenic amiloride-sensitive Na+ transport were examined in the distal colon of immature and adult rats. The decrease of the short-circuit current after amiloride addition (ISCamil) was used as a measure of electrogenic Na+ absorption. Plasma aldosterone levels were six to eight times higher between d 15 and 30 than in younger suckling or prepubertal rats. In adult rats, the plasma aldosterone was approximately 30 times lower than in young animals kept on the same standard diet. ISCamil followed the developmental profile of plasma aldosterone. ISCamil increased between d 10 and 20, reached a plateau between d 20 and 25, then decreased and entirely disappeared after d 30. ISCamil was zero in adult distal colon but was induced if dietary Na+ intake decreased below 100 mumol Na+/(100 g body wt.d) and plasma aldosterone increased above 200-300 pg/mL. Adrenalectomy, or high Na+ intake, inhibited ISCamil and significantly depressed plasma aldosterone in 20- and 25-d-old rats. Premature weaning decreased ISCamil without appreciable changes in plasma aldosterone in 18- and 20-d-old rats, prolonged suckling inhibited ISCamil and caused a significant depression of plasma aldosterone. We conclude from these results that the postnatal changes of distal colonic Na+ transport are regulated predominantly by circulating aldosterone and dietary Na+ intake. PMID- 7494660 TI - Age- and dietary-related distributions of hepatic arachidonic and docosahexaenoic acid in early infancy. AB - A dietary-related deficiency of docosahexaenoic acid [C22:6(n-3)] in infant cerebral cortex has been identified. Absence or very low rates of hepatic synthesis from the essential fatty acid precursor, alpha-linolenic acid [C18:3(n 3)], in early life may have been a contributory factor. We have analyzed liver total lipid fatty acid compositions in 27 term (37-42 wk gestation) and 4 preterm (30-33 wk gestation) infants who died within the first 6 mo of life from sudden infant death syndrome. The infants were fed exclusively either human or formula milks. Formula-fed infants were subdivided into two groups, one receiving SMA milk with an alpha-linolenic acid content at 1.5% of total fatty acids and the other a composite group fed milks with low alpha-linolenic acid concentrations (< 0.1% to 0.4%). The hepatic content of arachidonic acid [C20:4(n-6)] and docosahexaenoic acid was generally lower in both formula-fed groups than in the human milk-fed group. The age-related distributions of docosahexaenoic acid showed that coincident minimum levels were present in both formula groups in the third month of life. This may indicate that the hepatic enzymes involved in the final stage (delta 4-desaturation) conversion of alpha-linolenic acid to docosahexaenoic acid could be inactive in the first months of life. Emphasis must be on provision of preformed dietary docosahexaenoic acid and possibly arachidonic acid as well as their essential fatty acid precursors, to both term and preterm infants for at least the first 16 wk of life. PMID- 7494661 TI - Growth delay after liver transplantation in childhood: studies of underlying mechanisms. AB - After liver transplantation in children, growth is often impaired, but the underlying mechanisms are unknown. Glucocorticoids used for immunosuppression are believed to be partly responsible. After renal transplantation in children, reduced growth hormone (GH) secretion and increased serum insulin-like growth factor-binding protein-3 (IGFBP-3) levels have been reported. We attempted to find endocrine factors predicting growth in 18 prepubertal children followed for more than 1 y (mean 2.4 y) after liver transplantation. Spontaneous and stimulated GH secretion, serum IGF-I, IGFBP-3 concentrations, and endogenous cortisol production were measured. GH secretion was reduced in only two patients. Serum IGF-I concentration was normal, but serum IGFBP-3 was elevated or 1 SD above the mean for age in 62% of the patients. Endogenous cortisol production was reduced in most patients during the first year and improved later in only a few. Growth velocity after transplantation did not correlate with GH secretion, serum IGF-I or IGFBP-3 concentration, or with methylprednisolone dose, but correlated positively with serum basal (rs = 0.44, p < 0.05) and stimulated (rs = 0.53, p < 0.005) cortisol concentration. In conclusion, after liver transplantation 1) the normal pulsatile character of nocturnal GH secretion is sustained, and the GH response to stimulation is reduced in only a few patients; 2) serum IGF-I concentrations are normal; 3) serum IGFBP-3 concentrations are elevated or in the upper part of the normal range in most patients; and 4) endogenous cortisol production is reduced in most patients and correlates positively with growth velocity. PMID- 7494662 TI - Erythrocyte incorporation of iron by 56-day-old infants fed a 58Fe-labeled supplement. AB - In an effort to obtain information about absorption of supplemental iron by breast-fed infants during the early months of life, we determined erythrocyte incorporation of a stable iron isotope, administered to 56-d-old breast-fed infants in the form of a 58Fe-labeled vitamin-iron supplement. Infants of similar age fed a milk-based formula low in iron (approximately 4 mg/L) were also studied. The 58Fe-labeled vitamin-iron supplement was given between feedings. Fourteen days after administration of 58Fe, mean erythrocyte incorporation of the isotope was 7.8% of the dose by breast-fed infants and 4.4% of the dose by formula-fed infants. The feeding-related difference was statistically significant, probably reflecting the greater quantities of inhibitors of iron absorption in the intestines of formula-fed infants. With mean iron intake from the 58Fe-labeled vitamin-iron supplement of 7.99 mg for the breast-fed infants, erythrocyte incorporation of 7.8% of the dose corresponded to 0.62 mg, a value in the range of the estimated requirement for absorbed iron. We conclude that 2-mo old breast-fed infants are able to absorb nutritionally significant amounts of iron from an iron supplement. PMID- 7494663 TI - Insulin secretion and growth failure in uremia. AB - Glucose and insulin metabolism was studied in 16 adolescents with uremia on hemodialysis. Glucose tolerance was measured by i.v. glucose tolerance tests (IVGTT). Insulin sensitivity was measured by the euglycemic clamp technique. Insulin secretion during constant hyperglycemia was measured by the hyperglycemic clamp technique. Controls consisted of eight healthy young adults. These patients could be further subdivided into two groups with respect to their growth velocity SD score (GVSDS). Group one consisted of eight patients with GVSDS more than -2. This group had normal 1,25(OH)2D3 and demonstrated insulin resistance, hyperinsulinemia, and normal glucose tolerance when compared with adult controls. Group two consisted of eight patients with GVSDS less than -2. This latter group had low 1,25(OH)2D3, insulin resistance, but normoinsulinemia and glucose intolerance compared with adult controls. Published data in the literature showed that normal pubertal adolescents demonstrated insulin resistance and hyperinsulinemia compared with normal adults. Compared with normal values of insulin sensitivity in adolescents, both groups of patients on hemodialysis were still insulin resistant. The insulin secretion values in group one was similar to reported values for normal adolescents, whereas the values in group two were lower. GVSDS correlated significantly only with insulin secretion and not with glucose tolerance, insulin sensitivity, 1,25(OH)2D3, or PTH. Normal adolescents demonstrate an increase in insulin secretion as they go into puberty. It seems that the puberty growth spurt in adolescents both with normal health and renal failure may require increased insulin secretion as one of its hormonal requirements. PMID- 7494666 TI - Expression of cytokine receptors by human cord blood lymphocytes: comparison with adult blood lymphocytes. AB - The expression of receptors for several cytokines (IL 2, IL-4, IL-6, IL-7, tumor necrosis factor, and interferon-gamma) was examined in human cord blood cells in comparison with adult blood cells. A previously described high sensitivity immunofluorescence procedure was used to render the low levels of receptor measurable. Cord blood lymphocytes expressed measurable levels of most cytokine receptors, but expression tended to be lower than in adult blood cells. Examination of different lymphocyte subpopulations revealed a complex pattern with some cell types expressing particular receptors equivalent to adult levels. Cord and adult blood monocytes expressed similar cytokine receptor profiles. Receptor expression in cord lymphocytes could be regulated by activation. The results provide indications as to the relative activities of different cytokines in the development of the immune system in the neonate. PMID- 7494664 TI - Correlation of maternal cytophilic human immunodeficiency virus (HIV)-1 V3 loop peptide-specific antibodies in infants with vertical HIV transmission. AB - Approximately 70% of human immunodeficiency virus (HIV)-1-infected pregnant women do not transmit HIV to their offsprings. The identification of factors involved in maternal-child transmission of HIV is important for the design and implementation of therapeutic and prevention strategies. Recently we have developed a modified peripheral mononuclear cell (PBMC) culture system for in vitro antibody production (IVAP) by which we can distinguish serum-derived cytophilic anti-HIV-1 antibody from de novo synthesized antibody. In this study, we analyzed the presence of antibodies directed to HIV-1 gp160, gp120, gp41, and V3 loop synthetic peptides (C51, C53, and C57 from MN and IIIB strains) utilizing the grid-blot method in PBMC cultures of 52 mother-child pairs. Among the mothers (39 nontransmitters and 13 transmitters), presence of serum-derived cytophilic antibodies or de novo synthesized HIV V3 loop peptide-specific antibodies did not correlate with vertical transmission. However, PBMC-associated, cytophilic antibodies of maternal origin reactive with C51, C53, and C57 from MN and IIIB strains were identified in cultures of uninfected infants, but not in infected infants. These observations suggest that cytophilic antibodies of maternal origin directed to HIV-1 V3 loop peptides which are bound to infant cells might play a role in preventing vertical transmission. PMID- 7494665 TI - Increased expression of activation markers on CD8 lymphocytes in children with human immunodeficiency virus-1 infection. AB - The aims of the present study were to analyze the impact of perinatal human immunodeficiency virus (HIV)-1 infection on lymphocyte maturation in children, to determine the expression of activation markers on CD8+ cells, and to define predictors of survival in HIV-infected children. Seventy-one children presenting HIV-related symptoms were included in the study; 29 were less than 2 y old and 42 were 2 to 12 y of age. Results were compared with those obtained in normal children of a similar age. In HIV-infected children the proportion of CD4+ and CD8+CD45RA+ cells was significantly decreased, whereas that of CD8+, CD8+CD38+, and CD8+CD45RO+ cells was strikingly increased compared with controls. In children less than 2 y old the absolute number of CD4+ and CD8+CD45RA+ cells decreased, and the number of CD8+CD45RO+ cells increased significantly, whereas the number of CD8+ and CD8+CD38+ cells did not change. The absolute number of CD4+ T cells declined with age both among controls and among HIV-infected children. In contrast, the absolute number of CD8+ cells and CD8 subsets decreased with age only in controls but not in infected children. In HIV-1 infected children the expression of the CD38 and CD45RO markers on CD8+ cells was significantly correlated, indicating that these were activated cells. The survival of less than 2-y-old children with AIDS symptoms was positively correlated with the total number of CD8 cells and CD8+CD38+ cells at time of entry into the study.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7494667 TI - Immunologic effects of background prenatal and postnatal exposure to dioxins and polychlorinated biphenyls in Dutch infants. AB - Immunologic effects of pre- and postnatal polychlorinated biphenyl (PCB)/dioxin exposure in Dutch infants from birth to 18 mo of age are explored. The total study group consisted of 207 healthy mother-infant pairs, of which 105 infants were breast-fed and 102 children were bottle-fed. Prenatal PCB exposure was estimated by the PCB sum (PCB congeners 118, 138, 153, and 180) in maternal blood and the total toxic equivalent (TEQ) level in human milk (17 dioxin and 8 dioxin like PCB congeners). Postnatal PCB/dioxin exposure was calculated as a product of the total TEQ level in human milk multiplied by the weeks of breast-feeding. The number of periods with rhinitis, bronchitis, tonsillitis, and otitis during the first 18 mo of life was used as an estimate of the health status of the infants. Humoral immunity was measured at 18 mo of age by detecting antibody levels to mumps, measles, and rubella. White blood cell counts (monocytes, granulocytes, and lymphocytes) and immunologic marker analyses CD4+ T-lymphocytes, CD8+ T lymphocytes, activated T-lymphocytes (HLA-DR+CD3+), as well as T cell receptor (TcR) alpha beta+, TcR gamma delta+, CD4+CD45RA+ and CD4+CD45RO+ T-lymphocytes, B lymphocytes (CD19+ and/or CD20+) and NK cells (CD16+ and/or CD56+/CD3-) in cord blood and venous blood at 3 and 18 mo of age were assessed in a subgroup of 55 infants. There was no relationship between pre- and postnatal PCB/dioxin exposure and upper or lower respiratory tract symptoms or humoral antibody production.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7494668 TI - Measurement of fat-free mass in infants. AB - Body composition data are important for adequate monitoring of growth and nutritional status in infants. 18O Isotope dilution techniques (ID18-O) are widely used to estimate total body water (TBW) and calculate fat-free mass (FFM). A problem of isotope dilution is an underestimation of TBW by the extrapolation to t = O approach and an overestimation of TBW by the plateau approach. Using total body electrical conductivity (TOBEC) as the reference technique we validated the extrapolation approach by 149 measurements (boys, n = 76; girls, n = 73) in 50 healthy infants aged 1-12 mo. TOBEC-derived FFM and fat mass were in excellent agreement with Fomon's reference data. Strictly linear relationships with slopes not significantly different from one were found between FFM estimated by TOBEC (FFMTOBEC) and FFM estimated by ID18-O (FFM18-O) (r = 0.98 and residual SD = 0.29 for boys, r = 0.98 and residual SD = 0.32 for girls). FFM18-O was slightly but significantly lower than FFMTOBEC, the difference being on average 0.18 (+/- 0.24) kg for girls and 0.08 (+/- 0.21) kg for boys (i.e. respectively 4 (+/- 4.5)% (p < 0.0001) and 1.5 (+/- 3.9)% (p = 0.004) of FFMTOBEC). We conclude that ID18-O using the extrapolation to t = O approach is suitable for TBW and FFM estimations in groups of infants. Due to the considerable measurement error of ID18-O (estimated at approximately 6%), individual TBW18-O and FFM18-O estimates should be considered with some caution. PMID- 7494669 TI - Growth hormone (GH) treatment up-regulates GH receptor mRNA levels in adipocytes from patients with GH deficiency and Prader-Willi syndrome. AB - We have investigated the effect of growth hormone (GH) treatment on GH receptor mRNA expression in five prepubertal children with Prader-Willi syndrome and in eight patients with GH deficiency. An adipose tissue needle biopsy was taken before and after 2-4 mo of GH treatment, and RNA was isolated from adipose tissue and from adipocytes. GH receptor mRNA levels were determined by an RNase protection/solution hybridization assay. To further assess the specificity of the assay for GH receptor mRNAs, RNA extracted from human adipose tissue was subjected to Northern blot analysis. GH treatment significantly increased GH receptor mRNA levels in adipose tissue and isolated adipocytes. Our results indicate that GH may have an important role in regulating the GH receptor in humans. PMID- 7494670 TI - Dietary fiber in weaning foods of young children. AB - OBJECTIVE: An evaluation of the opportunity to introduce fiber in the diet of weaning infants. METHODS: A descriptive review of the literature to assess the role of fiber in the following dietary interventions: (1) definition of the nutrient amounts that meet the weaning infant's physiologic needs; (2) modulation of digestive and absorption processes; and (3) improving the nutritional balance between the 6th and the 12th months of life and setting habitual dietary patterns for subsequent years. FINDINGS: Whole cereals, nonstarchy vegetables, fruits, and legumes lower the caloric and proteic density of meals, modulate nutrient and antigen absorption, and provide bulk material. They supply proteins of low biological value, minimal amounts of lipids (mostly essential polyunsaturated), complex carbohydrates, and soluble fiber, which are fermented into short-chain fatty acids by the colonic flora, and insoluble fiber that modulates intestinal function. Minerals, trace elements, and vitamins add to the value of fiber containing foods in the diet. Recent nutritional surveys indicate that the diets of 12-month-old infants tend to include too much animal proteins and to be too protein-dense, which could be corrected with an increase of fiber-containing foods. CONCLUSIONS: Fiber-containing foods share unique characteristics with nutritional and metabolic implications for the weaning infant. Whole cereals, green vegetables, and legumes should be routinely introduced during the weaning process to achieve a better nutritional balance and to accustom children to diets with fiber content. The daily intake of fiber should be gradually increased to 5 g/d during the second semester of life. PMID- 7494672 TI - The role of dietary fiber in the development and treatment of childhood obesity. AB - Childhood obesity is viewed as a public health problem in the United States because of its assumed high prevalence and increasing secular trend. The best estimate of the genetic contribution to obesity ranges from 5% to 25%. Environmental factors play a major role in obesity development. Low income and a low level of education have been associated with obesity, particularly among white women. Caloric intake as a risk factor for obesity has not been clearly established. This lack of a clear-cut association may be attributable to the problem of accurately measuring caloric intake. Several studies have linked increased total fat intake, rather than caloric intake, with obesity. Some studies have linked television viewing to obesity in children. Obesity is rare among the populations of developing countries, where dietary fiber intake is high. Explanations for the role of dietary fiber in obesity include a reduced caloric density of the foods, a slower rate of food ingestion, and possible effects on satiety. Most studies on the role of fiber in the treatment of obesity have been somewhat limited by lack of comparison groups, inadequate sample sizes, and short durations of the observations. However, although limited, the available evidence suggests that fiber potentially could play a useful role in weight reduction. For children, fiber administration should be considered as an adjuvant therapy rather than a primary modality, because fiber might aid in promoting satiety during meals and curbing hunger between meals.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7494671 TI - The role of fiber in the treatment of hypercholesterolemia in children and adolescents. AB - The effect of adding water-soluble fiber to a diet low in total fat, saturated fat, and cholesterol to treat hypercholesterolemic children and adolescents with elevated plasma low-density lipoprotein (LDL) cholesterol levels was assessed. In more than a half-dozen studies, the effect of water-soluble fiber on the LDL cholesterol level ranged from no change to as high as a 23% decrease using oat bran, psyllium, or locust bean gum. The wide range of effects in these studies may be related to the quality of the dietary intervention or to different methods of randomization, blinding, dietary assessment, and laboratory measurement. For example, the addition of supplemented soluble fiber (psyllium) to a step 1 diet may provide additional lowering of LDL cholesterol of 10% to 15%. However, in children consuming the more stringent step 2 diet, the addition of water-soluble fiber may have less additional effects on LDL cholesterol. As recommended by the National Cholesterol Education Program Expert Panel on Blood Cholesterol Levels in Children and Adolescents, dietary therapy, that is, a diet low in total fat, saturated fat, and cholesterol, remains the cornerstone of treatment for children and adolescents with elevated LDL cholesterol levels. The use of foods high in water-soluble fiber that contain no cholesterol and are low in saturated fat remains a good choice in children following a step 1 or step 2 diet. Additional clinical trials in larger numbers of well-defined subjects will be needed to assess further the utility of adding water-soluble fiber supplements to the National Cholesterol Education Program step 1 or step 2 diets.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7494674 TI - Dietary fiber for children: how much? AB - BACKGROUND: Dietary fiber intakes of most American children are lower than current American Academy of Pediatrics recommendations. Intakes of vegetarian children come closer to these levels. RESULTS: We summarize dietary fiber recommendations for children based on existing evidence. The general public needs guidance on appropriate fiber intake levels for children and adolescents. It is important to ensure that energy intakes are adequate by monitoring child weight, growth, and size, especially when fiber intakes are very high. At levels of "age plus 5 g" there seem to be few problems. CONCLUSIONS: Age plus 5 g is a reasonable recommendation and is easier to remember than others by weight or energy level, although it never exceeds them. Age plus 15 g is clearly excessive; there is less evidence about ill effects arising with age plus 10 g. Delivery of dietary fiber in food rather than by supplements is suggested to ensure intakes of other nutrients and to avoid medicalizing dietary intake. In addition to dietary fiber recommendations for the general population of healthy children, individualized recommendations may be necessary for some high-risk groups. Practical steps to increase child fiber intakes from food sources are provided. The article concludes with suggestions for further research. Age plus 5 g is a reasonable minimum recommendation for dietary fiber intakes for children older than 3 years of age. PMID- 7494673 TI - Is a high-fiber diet safe for children? AB - OBJECTIVES: Although dietary fiber is associated with important health benefits in childhood, there have been concerns that very high fiber diets may result in adverse health effects. This report reviews the major safety concerns associated with consumption of very high fiber diets, estimates the amount of fiber that may cause adverse physiologic effects in children, and proposes safe levels of dietary fiber intake for children and adolescents. METHODS: Published studies on dietary fiber intake in childhood were reviewed to determine major safety concerns, to document adverse effects, to characterize subjects involved and the dose and type of fiber consumed, and to estimate potential relevance to US children and adolescents. Levels of dietary fiber reported to have adverse health effects were compared with recommended levels of fiber intake for children older than 2 years of age. RESULTS AND CONCLUSIONS: A review of the scientific literature suggests that a small loss of energy, protein, and fat may occur with a high intake of dietary fiber. However, this small loss of energy is unlikely to be significant to children consuming adequate levels of major nutrients, especially at conservative fiber intakes as recommended by the American Health Foundation's age plus 5 formula. In addition, it is estimated that even with a doubling of current dietary fiber, there is unlikely to be an adverse effect on serum vitamin and mineral concentrations in healthy US children consuming a balanced diet containing adequate levels of nutrients. Thus, evidence suggests that for US children, a moderate increase in dietary fiber is more likely to be healthful than harmful. PMID- 7494675 TI - A summary of conference recommendations on dietary fiber in childhood. Conference on Dietary Fiber in Childhood, New York, May 24, 1994. American Academy of Pediatrics. PMID- 7494676 TI - The role of dietary fiber in childhood. Proceedings of a conference. May 24, 1994, New York, New York. PMID- 7494677 TI - A new recommendation for dietary fiber in childhood. AB - OBJECTIVE: The consumption of dietary fiber in childhood is associated with important health benefits, especially with respect to promoting normal laxation. Dietary fiber also may help reduce the future risk of cardiovascular disease, some cancers, and adult-onset diabetes. At present, there are few specific guidelines for dietary fiber intake in childhood. Our goals were to review the benefits and risks of dietary fiber in childhood and to propose a safe and effective quantitative recommendation for the US pediatric population. METHOD: Current intake of dietary fiber in childhood was reviewed, including data from the US Department of Agriculture Nationwide Food Consumption (1987-1988) and National Health and Nutrition Examination II (1976-1980) Survey. Current intake was compared with existing fiber recommendations, including the 0.5-g/kg guideline proposed by the American Academy of Pediatrics Committee on Nutrition. Recommended fiber intake was reviewed with respect to levels required for specific health benefits, as well as levels that may result in adverse health effects. RESULTS AND CONCLUSIONS: A new recommendation for dietary fiber intake was developed, based on the age of the child, health benefits, and safety concerns. We recommended that children older than 2 years of age consume a minimal amount of dietary fiber equivalent to age plus 5 g/d. A safe range of dietary fiber intake for children is suggested to be between age plus 5 and age plus 10 g/d. This range of dietary fiber intake is thought to be safe even if intake of some vitamins and minerals is marginal, should provide enough fiber for normal laxation, and may help prevent future chronic disease. PMID- 7494678 TI - Dietary fiber intake of children: the Bogalusa Heart Study. AB - Age and secular trends in dietary fiber intake of children were examined using the 24-hour dietary recall method. Dietary fiber intake of five cohorts of 10 year-olds (n = 1254); two cohorts of 13-year-olds (n = 360); and children ages 10 (n = 767), 13 (n = 249), 15 (n = 108), and 17 (n = 89) years was examined from 1976 to 1988. Total dietary fiber intake remained unchanged from 1976 to 1988, even after adjusting for energy intake; averaging 12 or 5 g/100 calories (kcal). Black children had higher fiber intakes per 1000 kcal than white children from ages 10 to 17 years. Consumption of vegetables and/or soups and breads and/or cereals accounted for 53% (10 years) to 70% (13 years) of the total fiber consumed. When children were stratified into quartiles based on fiber intake per 1000 kcal, the percent of energy from total fat and saturated fat was lower, and the percent of energy from carbohydrates was higher in children with higher fiber intakes per 1000 kcal. Additional studies are needed to document dietary fiber intakes of children and their impact on overall diet composition and cardiovascular risk factors. PMID- 7494679 TI - Fiber in the diet of US children: results of national surveys. AB - OBJECTIVE: Because fiber intake among children in the United States is poorly documented, our objectives were to determine intakes and sources of fiber in the diets of children 2 to 18 years of age and to examine the trends. METHODS: We used the US Department of Agriculture's 1977 to 1978 and 1987 to 1988 Nationwide Food Consumption Surveys for our analysis. Children were classified by age and gender groups, 2 to 5 years, 6 to 11 years, boys 12 to 18 years, and girls 12 to 18 years. The percentages of children meeting the American Health Foundation's fiber recommendations for children were also determined. RESULTS: Analyses of the US Department of Agriculture's 1977-1978 and 1987-1988 survey data indicate that (1) large percentages (55% to 90%) of children are not meeting minimum fiber intake recommendations established by the American Health Foundation; (2) mean fiber intakes declined between 1977-1978 and 1987-1988; (3) fiber from fruits and vegetables declined during this period; and (4) fiber from grains, especially from ready-to-eat breakfast cereals, increased significantly. The results also show that eating breakfast contributes to meeting daily fiber recommendations. CONCLUSIONS: Given the importance of fiber in promoting optimum gastrointestinal function in childhood and in treating chronic diseases such as heart disease, diabetes, and cancer in adulthood, the trends documented in this article are disturbing and suggest that efforts are needed to encourage the consumption of fiber-rich foods among children. PMID- 7494680 TI - An overview of the effects of dietary fiber on gastrointestinal transit. AB - This article is a review of the effects that ingested dietary fiber has on gastrointestinal transit time. The various phases of gastrointestinal motility are considered, and the effect of fiber on these various components is explored. Fiber affects each phase of gastrointestinal motility differently; however, the most dramatic effect on decreased transit time and frequency of bowel movements result from the variations it causes in colonic transit time. The mechanisms of defecation in children and the ways in which fiber affects the frequency of bowel movements are emphasized. All types of fiber do not affect gastrointestinal transit in a similar manner, and different preparations of the same fiber either may delay or may increase the time of intestinal transit. In general, fiber is found to increase the frequency of bowel movements and may prove to be of considerable benefit in treating constipation, a common childhood disorder. PMID- 7494682 TI - Principles of adult learning: how can they help. PMID- 7494681 TI - Constipation and dietary fiber intake in children. AB - OBJECTIVE: Severe constipation in children is associated with rectal overdistension and insensitivity. Treatment typically involves three phases: disimpaction (days to weeks), laxative use (months), and a high-fiber diet (lifelong). The purpose of this survey is a discussion of therapies that recognize the unique problems that children with severe constipation have. METHODS: Four-day diet logs were obtained from children who had no history of chronic bowel disease. Their intake of dietary fiber was determined from the logs and was compared with the dietary fiber ingested by children who had chronic constipation. These data sets were compared in light of our current understanding of the need for dietary fiber. RESULTS: Approximately half of the children from families who were health conscious enough to request dietary evaluation still fell below the age + 5 guidelines for grams of dietary fiber intake per day. The children referred to use with chronic constipation had all been instructed "to eat a high-fiber diet." Those constipated patients were consuming less than one fourth of the recommended fiber intake. CONCLUSIONS: This survey underscored the difficulties in beginning and in maintaining high-fiber diets in children. When families receive advice to administer a high-fiber diet, they are unable to accomplish this unless they receive intensive and ongoing dietary counseling. Even among health-conscious families, only half of the children received the recommended amounts of dietary fiber. Further public education in this regard is warranted. PMID- 7494683 TI - Combating the hidden enemy. PMID- 7494684 TI - Nursing resources on the Internet. PMID- 7494685 TI - Commissioner of Health announces new HIV committee: infected health care workers is the focus. PMID- 7494686 TI - The Patient Protection Act: A-2928. PMID- 7494687 TI - The narrative enlarging. A biography of the community college--nursing home partnership project. PMID- 7494689 TI - Friends for life and death. Interview by David Payne. PMID- 7494688 TI - Raising the profile of the school nurse. PMID- 7494690 TI - Nowhere to turn. PMID- 7494691 TI - On duty. Interview by Allan Jabez. PMID- 7494692 TI - Men behaving badly. PMID- 7494693 TI - Lights, camera, action. PMID- 7494694 TI - A cognitive approach to panic disorder. AB - Panic disorder can be a chronic and disabling problem, with sufferers frequently using of medical and psychiatric services. In this paper, the nature of panic attacks and panic disorder is outlined, and a cognitive model of panic disorder described. Treatment is outlined, and a case example given. PMID- 7494695 TI - Management of anxiety states. AB - Anxiety states affect millions of people in the UK and as such constitute a major public health problem. Specialist mental health nurses can provide very effective treatments for a range of some of the most severe of these conditions. However, because of the prevalence of anxiety, all nurses should know something of their nature and management. PMID- 7494696 TI - Be honest about cuts. PMID- 7494698 TI - The adrenal glands. PMID- 7494697 TI - Perceptions of childbirth: are women properly informed? AB - This paper is the second of two examining the differing perceptions of childbirth between women and medical professionals. It has been suggested that the differing views have led to a position where medical knowledge and frames of reference are accepted and legitimated by a system which leaves women feeling alienated and dissatisfied with the conduct of delivery. This paper takes the discussion forward by reviewing the recommendations of the Changing Childbirth document, asking whose responsibility it is to inform women about their condition and treatment, examining the importance of continuity of care to the information sharing process, the impact that this could have on women's ability to exercise informed choice and, ultimately, assessing whether change is possible. PMID- 7494699 TI - Caught in the frontline. PMID- 7494700 TI - Living with dementia. PMID- 7494702 TI - Herbalism: useful remedies. PMID- 7494701 TI - Stress reduction. PMID- 7494703 TI - Professional development. Trauma. The role of the nurse. PMID- 7494704 TI - Leading from the front. PMID- 7494705 TI - Knowledge for care. PMID- 7494707 TI - Patient Education 2000. Proceedings of a congress. Geneva, 1-4 June 1994. PMID- 7494706 TI - Assessment on the record. PMID- 7494709 TI - Bridges, why and from where to where? AB - Therapeutic intervention for chronic diseases does not rely only on drugs but is also strictly dependent on how the patient has been informed about his disease and how he is able to master the various skills required by his treatment. Education therefore plays a fundamental role in the efficacy of control of those diseases. But what is education? Among the various possible definitions, one could be metaphorically illustrated by bridges. How to get the message across from the medical world to that of the patient, or, inside the medical professions from one group of providers to another. Different bridges may have different functions or roles. One is for commuting in the current daily life. Another one is to ensure vital needs. Bridges help also to discover new territories, but any new visitor may not always be welcomed in these new regions. Bridges need to be solidly constructed with a structure which can be methodologically described. This may help repair in case of problems. Patient education is a kind of bridge submitted to all sorts of forces and barriers which may interfere with the passage of knowledge and skills from the health care providers to the patient and his family. PMID- 7494708 TI - Coping with a chronic disease: the role of the mind and spirit. AB - This article presents personal experience of surgeons with the rehabilitation of disabled people. Attention is paid to the role of mind and spirit in the rehabilitation with particular attention to people with leprosy. The consequences for patient education, emotional support and attitudes of professionals are described. The influence of hope and faith in the case for disabled people is stressed. PMID- 7494710 TI - Why should patients do what we ask them to do? AB - Many factors contribute to patients' self-management success. In the main these are attitudes, beliefs or perceptions of the patients rather than demographic characteristics. Patients do need not only to acquire knowledge and self management abilities, but also the desire to apply them on a life-long basis. They require understanding and support. Education is a life-long process. All those involved in care need training in the educational process. Both the targets set and achieved by the patients need regular assessment. It is critical to perform a comprehensive assessment of the influencing factors. Conclusions are drawn on the consequences for relief education. PMID- 7494711 TI - Teaching, training or educating patients? Influence of contexts and models of education and care on practice in patient education. AB - This paper presents the rationale, theoretical developments and operationalization of the role of contexts of care and education in practice, choice of methods and effects of patient education in chronic diseases. Starting from a review on the evolution of models of care, education and health education, and on the influence of characteristics of organization of care, it proposes an analysis model of contexts of care and education. A study carried out in 1993 in 10 Belgian and northern France hospitals, with 400 patients, is presented. Simultaneously, pre-post test evaluations of the effects of current patient education work on 40 compliance factors, and in-depth analyses of contexts of care and education were undertaken. The possible relationship between variety and number of effects of education and type of care and education is examined and discussed. PMID- 7494712 TI - Barriers and challenges in training health care providers for patient education. AB - Patient education and chronic illness are linked inextricably in the training of health care providers as we move to the year 2000. However, professional training remains organised around the traditional clinical interview with its focus on acute illness. Research into aspects of the health care consultation is well advanced, but we are only just beginning to understand the implications of the long-term interaction between patient and provider in the management of illnesses which cannot be cured. The challenges for health care providers are: to relinquish control of the consultation when it is appropriate; to be aware of the human biases affecting clinical judgment; to develop a professional approach to the human aspects of their relationships with patients and other professionals as part of a management team; and to focus on the longer term relationship and long term benefit to the patient. PMID- 7494713 TI - Empowerment: from philosophy to practice. AB - Our paper reviews the philosophical and theoretical foundations of the empowerment approach to chronic disease care and education. The fundamental differences between the compliance and empowerment approaches are elucidated. The empowerment philosophy is based on the premise that human beings have the capacity to make choices and are responsible for the consequences of their choices. Empowerment is defined as an educational process designed to help patients develop the knowledge, skills, attitudes, and degree of self-awareness necessary to effectively assume responsibility for their health-related decisions. Using empowerment as an intellectual foundation, an holistic approach to health education is presented. Topics such as well-being, self-image, motivation, adaptability, stress management, problem-solving, social support, self-awareness, and hope, are discussed. Our paper asserts that health educators have a responsibility to address the physical, emotional, cognitive, and spiritual needs of persons challenged to live with a chronic disease. PMID- 7494714 TI - Coping-based intervention strategies. AB - Coping is explained as a circular process involving three repetitive steps: (1) perception of a change in well-being; (2) cognitive processing of information, in assessing its impact and possible consequences; and (3) coping as such, including many different modes. In general, active, task-oriented behaviour in which patients tackle problems, remain optimistic and seek attention and care is considered favourable. A passive and withdrawn attitude with ruminations, self accusation and resignation is known to be unfavourable. Based on problem-solving theory, a coping-based intervention strategy is presented as follows: (1) establishing a therapeutic relationship; (2) analysis of the problem (situation and actual coping); (3) definition of problem for patient; (4) defining goals; and (5) working through and modification of coping. A rich body of scientific evidence now exists in almost all areas of medicine that such short and structured interventions contribute significantly to the healing process. PMID- 7494715 TI - Illness and culture: learning differences. AB - A review is presented of several aspects of illness and culture. Stressing the importance of suffering in life, attention is paid to the influence of the image of the world on the relationship to the disease and the doctor. This is illustrated by a belief in fate. Further theoretical concepts from Kleinman, Helman, Schweder and Sontag are applied to medical situations. Cultural differences may be much more important for the treatment of patients than acknowledged until now. PMID- 7494716 TI - Totemic medicine among the American Indians of the northwest coast. AB - A description is presented of the medical beliefs of the totemic people on the Northwest American coast. The principles of Shamanic therapy are analysed. The consequences for therapeutic views by medical schools are discussed. PMID- 7494717 TI - The body image in health and disease: exploring patients' maps of body and self. AB - Patient education in chronic diseases should always take into account the many maps--individual, cultural and medical--that patients have of their own bodies, in both health and disease. Physicians should become explorers, working in co operation with patients and their families to understand their perceptions of body and self--and whether or not these are compatible with medical interventions. PMID- 7494718 TI - Home dialysis: lessons in patient education. AB - The concept of home haemodialysis for the regular treatment of renal failure was first put into practice in 1964, 30 years ago. It proved extremely successful. This paper describes the epidemiology of renal failure, the reasons for home haemodialysis and the educational role necessary to ensure its success. Continuous ambulatory peritoneal dialysis (CAPD), or autonomous dialysis, is a self-care peritoneal treatment which developed after 1978 for a wider age range of patients. Its rapid expansion needed a professional approach to patient education. Also discussed are the problems patients encounter with conceptual skills, as opposed to the easy acquirement of practical skills and the paper demonstrates how persons, without formal medical and nursing education, can master complex treatment skills. PMID- 7494719 TI - A community-based public health initiative integrating a traditional war canoe culture within today's contemporary lifestyles: the 'C.E.D.A.R.' project. AB - How many people have experienced the thrill of a competition that spans generations upon generations and is distinctly unique to the Northwest Coastal Salish Tribes. War canoe racing still exists today and is a constant reminder of a way of life that is quickly vanishing, almost as fast as the ancient forests of cedar--the tree of life for the Northwest Tribes. The C.E.D.A.R. Project's overall mission is 'to improve the health of the Lummi community.' The goals of the workplan seek collaboration of the partners and institutional change; increased health related education and health professions opportunities; and empowerment of the community partners to assure the sustainability of the Community-Based Public Health Initiative. PMID- 7494720 TI - Why bother educating the multi-disciplinary team and the patient--the example of prevention of lower extremity amputation in diabetes. AB - Amongst all the long-term complications of diabetes it is believed that foot ulceration and amputation are the most preventable: small studies have suggested that more than 50% of amputations should be avoidable by screening and appropriate education. There is ample evidence that simple non-invasive screening tests are not being carried out in many diabetic clinics--one study confirmed that poor patient care was responsible for 50% of heel ulcers in a hospital setting. There is a similar lack of awareness of potential problems in many 'at risk' diabetic patients. Thus, simple screening of diabetic patients should be routine clinical practice: those identified as being at 'high risk' of foot, problems should receive appropriate education in protective foot care. The rewards are potentially great, and include the realization of the St. Vincent target, a 50% reduction in amputation. PMID- 7494721 TI - The use of group programs to improve medication compliance in patients with chronic diseases. AB - Non-compliance with medication is one of the major factors in the failure of therapeutic programs for patients having a chronic disease. The good results of a controlled study on the evolution of schizophrenic patients, who participated in a medication group program aimed at diminishing affective components of negative attitudes towards neuroleptics, are discussed. The building up of specific questionnaires could help in understanding the nature of the prejudices towards the use of medication in patients with chronic diseases. The organization of group programs could diminish patients' prejudices, thus improving compliance. PMID- 7494722 TI - Three years' experience after national introduction of teaching programs for type II diabetic patients in Germany: how to train general practitioners. AB - Beginning in July 1991, office-based physicians in Germany were remunerated for providing a structured treatment and teaching program to non-insulin-treated type II diabetic patients. The treatment and teaching program is based upon the respective recommendations of the German Diabetes Association. This article describes the structure of the program and the courses for general practitioners and their nurses. The prospective studies proving the beneficial effects of the program are summarized. PMID- 7494723 TI - Coping with employment discrimination against diabetics: trends in social medicine and social psychology. AB - In many European countries, social-medical aspects in the management of diabetes mellitus are not satisfactorily respected. Our contribution reports a study addressing the impact of diabetes on the patient's career and daily work, in order to determine the extent to which diabetics are being discriminated against at work. Type I diabetics were questioned about their experience, and not on the objective burden. A questionnaire was developed to evaluate patients' social and employment problems. Few elements of an education program for Type I diabetes optimizing social skills (social competence) are demonstrated. In a group of 6-8 patients, assertive behavior in the work place is modelled (e.g., for hypoglycaemia, social phobia) by applying psychological methods (behavior modification role-playing). These methods can help diabetic patients to master their discrimination. They learn assertive behavior in social situations with superiors and colleagues and develop self-confidence (self-efficacy). This special education program supports Type I diabetics in coping with employment discrimination. PMID- 7494724 TI - Diabetes service management training and the need for a patient perspective: a 10 year evolution of training strategies and goals. AB - In 1977, the Swedish National Board of Health and Welfare published guidelines for comprehensive care of patients with diabetes. The Stockholm County (1.6 million inhabitants) responded and opened a Diabetes Education and Training Centre, LUCD, in 1979. The goal was to reduce the impact of long-term complications. The centre should translate available knowledge about diabetes care into clinical practice, promoting a shift of patients and resources to the primary health care sector. Follow-up studies and new knowledge have changed the short-term training strategies; the long-term goal has remained unchanged. Current work is concentrated on training of staff, improving methods to support better patient learning, aiding organisational changes, and supporting large scale interventions aimed at both patients, staff and the public. The patient can realize the goals. Knowledge and demands expressed by persons with diabetes are necessary for improvements of diabetes care. The skilled patient is also a guarantee for continuity of care. Regular follow-up of diabetes services was an important impetus for progressive change of short-term training strategies. PMID- 7494725 TI - Self, social structure, and health-promoting behavior in hypertensive patients. AB - A description is presented of views on coping with stress due to chronic, socio emotionally distressing conditions in the course of hypertension. Four theoretical approaches are discussed, and illustrated by recent studies: 1. lack of support, 2. relative deprivation, 3. job strain, 4. effort-reward imbalance. The implications for stress management and patient education are discussed. PMID- 7494727 TI - Population-based management of diabetes care. AB - Meeting the needs of patients with chronic illness in usual primary care practice is difficult. Group Health Cooperative of Puget Sound (GHC), an HMO serving about 400,000 people in western Washington state, is using the strategy of population based management of care to improve care and outcomes for its 13,000 diabetic patients. Population-based care uses guidelines, and epidemiologic data and techniques to plan, organize, deliver and monitor care to specific clinical sub populations such as diabetics. Under this approach, guidelines for care and the outcomes effected by those guidelines are defined. Current practice is reviewed, problems are identified, and a new intervention plan devised and put into place. Performance and outcomes are monitored and the plan revised accordingly. Population-based care must occur both centrally to provide guidelines, data, new interventions and other system support, but more importantly at the level of the individual medical practice if it is to reach all patients in the population. In this paper, we describe both central and practice-based efforts at GHC to apply population-based care to improving outcomes for diabetic patients. PMID- 7494726 TI - A community-wide promotion of asthma self-management in New Zealand. AB - Asthma affects approximately 9% of young adults in New Zealand, with evidence of an increase in prevalence over recent years. Morbidity and mortality from this condition have caused widespread concern. In response to identified deficiencies in asthma management, initiatives to promote self-management using an asthma action plan were promoted throughout the Canterbury region in 1991 as part of a regional pilot study. A resource kit was prepared and self-management principles were promoted to general practitioners in discussion workshops. A community-wide promotion of the action plan was followed by a process and outcome evaluation. The plan and associated resources were well received by doctors and patients. Seventy-seven percent of doctors responding to a questionnaire had used the plan, and of these, 94% indicated that they would continue to use the plan in the future. Patients who obtained a completed plan experienced a greater reduction in nocturnal waking with asthma and improved self-management skills. PMID- 7494729 TI - From past experiences to the promotion of current resources. AB - This paper notes the development of diabetes nurse specialists and recommends the creation of nurse specialist diabetic clinics. It considers the current manpower resources and how they may be used in the provision of diabetes education and care. Experience of training lay advisors is reported. PMID- 7494728 TI - The role of sickness funds in secondary and tertiary prevention. AB - Present Swiss federal legislation on health insurance provides only a very limited basis for an active engagement of sickness funds in prevention. Nevertheless, sickness funds have offered supplementary insurance coverage benefits for primary and secondary prevention. With the revision of the health insurance act the engagement of sickness funds in prevention is bound to change. With health promotion, medical prevention and rehabilitation all three levels of prevention will be considered in the revised act. In the context of the ongoing political discussion on cost containment in health care and on health insurance premium increase, an active engagement in prevention will be of high interest for sickness funds. Sickness funds have been active in patient education for many years at the institutional level. New fields of activity can be the promotion and probably also the installation of programs in patient education, ambulatory rehabilitation, and followup activities for individual patients and target populations. PMID- 7494730 TI - Educating the health care team. AB - How can health professional teams provide patient-centred care in the treatment of chronic disease? What education and training is needed to achieve this? Patient-centred care means helping people living with a condition make informed choices to maximise their quality of life. The health professional's role is to facilitate, to ensure patients and those around them take effective and appropriate self-management of the condition. To achieve this, high quality team working is needed amongst people who do not interact in the course of their day to-day work. Education for this should begin with students, expand when staff are newly qualified, become a feature of specialist training, and extend through continuing professional development. A unified philosophy and strategy then is needed for life-long professional education and training. Generalised principles are proposed for what should happen and who can best provide it, and these can be widely applied in many fields of health professional education. PMID- 7494731 TI - The role of the pharmaceutical industry in promoting patient education. AB - The role of the pharmaceutical industry in patient education consists of information related to products, information related to disease states and stimulation of all parts and partners in patient education. The 'package inserts' have advanced far beyond basic dosage prescribing information and precautions. Audiovisual instruction aids and telephone hot lines are increasingly used. Information on disease states are mainly distributed through the health care providers and patient organisations. They range from tools for special provisions, e.g. foot care in diabetes, up to complete systems for patient education. The stimulating activities range from the funding of education funds or conferences to the organisation of courses for health care professionals up to close cooperation with non-governmental and governmental organisations involved in patient education. The persons involved from industry became highly respected, fully integrated partners in the development of patient education. It is hoped that the commitment of industry in this issue might continue. PMID- 7494732 TI - Patient education: lessons from epilepsy. AB - Throughout the world some 40 lay associations are in the business of enabling patients with epilepsy to come to an understanding of their condition. This article aims to present the bones of this collective experience by detailing the positive outcomes that can be achieved for patients, for health professionals and for society in general, if patient education is delivered in a way that promotes confidence building, ensures that the educational locus is centred on the patient and enables the patient to be a partner in any decision-making process. This article will also suggest general guidelines that should underpin any patient education programme, as well as specific content. The questions of who should be responsible for delivering the programme and how patient education programmes can in reality become an integral part of health care plans are also addressed. PMID- 7494733 TI - Interprofessional cooperation: why is partnership so difficult? AB - The present state of interpersonal cooperation from the perspective of nursing is analysed, and some explanations are presented. The history of nurse-doctor cooperation in hospital is described. Examples of professional cooperation concerning the care of geriatric patients are also presented. PMID- 7494734 TI - Empowerment techniques: from doctor-centered (Balint approach) to patient-centred discussion groups. AB - The importance of training in the emotional domain has been stressed since 1950. Balint and his co-workers advocate the conduct doctor-centred seminars (training and research) for better understanding of the contact with patients. The discussions form an understanding of the content of the consultation within the context of the sense and relationship. Balint's method, with its emphasis on the role of psychology in medical care and on the interpersonal relations between the group members and their patients, is a satisfactory answer to the need for communication in the dispensing of health care. The case reports and group discussions attempt to throw a light on the doctor-patient relationship through the identification of its form and content. This diagnosis enables the physician, at times, to intervene therapeutically. An intervention is therapeutic when it brings to the fore an emotional or cognitive factor in the doctor-patient relationship and enables the patient to recognize something new about himself, and in doing so, even to 'change'. The ability to be acquired largely comprises that which is necessary for developing and understanding a doctor-patient relationship. At the same time family groups and psychosocial problems are taken into account. Junior Balint Groups for medical students have now been in existence at various universities. For 25 years general practitioners, clinic doctors, university staff and medical students have met for international seminars in Ascona, known under the name of 'Ascona Model' (WHO) with the intendion of adding medical training and consist mainly of Balint teamwork.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7494735 TI - What do patients really want to say to us, the health care providers?--Some answers from a nurse. PMID- 7494736 TI - Interdisciplinary and intersectoral approach: a challenge for integrated care. AB - Integrated care for health disorders, particularly chronic diseases, is a long term and complex challenge, particularly because of the involvement of many individuals with different beliefs, attitudes, assumptions and reward structures. Two basic conceptual models of disease--the biomedical and psychosocial--underlie many of these differences. The biomedical model views humans as the sum of multiple individual 'subsystems,' and disease represents dysfunction of one or more of these subsystems. This model is 'reductionist' and 'individualistic' in nature in that if 'THE' defective subsystem can be identified, studied and improved, it is assumed that health would return. The biomedical model focuses primarily on the individual with ill-health and has added greatly to our basic understanding of disease processes. The psychosocial model is 'interactive' and dynamic, and sees the 'whole' as more than the sum of its parts. This model values elements outside of the individual, e.g. work and home environment, as important in maintaining or establishing health. Because of fundamental differences between these 2 models of health and disease, conflicts, e.g. efficacy vs. exposure; role of individual vs. environment; etc., may exist among varying professionals regarding the nature, purpose, targets, structure, and consequences of integrated care programs. These fundamental conflicts, if unrecognized and ignored, can significantly attenuate the benefits of well intentioned prevention and treatment integrated care programs. PMID- 7494737 TI - What do patients really want to say to us, the health care workers? Some possible answers from a lay association. AB - From a personal point of view as diabetic and board member of diabetes associations in UK and Europe, attention is paid to the need for counseling and education. The role of patient associations in education is underlined, stressing also the negative aspects of the social position of diabetics in several European countries. PMID- 7494738 TI - The role of lay associations: difficulties encountered. AB - A review of Parkinson's disease stressing medical and psychosocial aspects is presented. The importance of the role of lay associations is described, paying particular attention to future developments. The foundation of an information bank is stressed as well as the cooperation with professionals to ensure standards of care and providing on-going patient education. Dutch initiatives are described as positive signs of the growing strength of lay organizations. PMID- 7494739 TI - Future perspectives of lay organizations. AB - The article describes the contributions of lay organizations in the field of diabetes. Attention is paid to the often-used terminology like laymen, patients and chronic disease. Further, the major contribution of lay organizations is stressed--their unique knowledge advocacy group for social rights, improvement, and partnership, also concerning the relationship with professionals. Financial restraints for the organization of people with chronic diseases are presented. Conclusions are drawn on the future role of lay organizations. PMID- 7494740 TI - The history of diabetes lay associations. AB - With much of the developed world having the increasing sociologic and economic problems of an aging population, there is an increased awareness of educating patients to not only understand and follow the instructions of their medical teams but to have increased responsibility for their own care. Of all the chronic conditions, diabetes, for which there is no cure, is among the most expensive and education-intensive. The lifelong Type I insulin-dependent diabetics need to know how to adjust their life-style and activities but also how to take and adjust several times daily insulin injections, diet and exercise and how to keep a wary eye to avoid complications that might be life-threatening. Since individual instruction is not possible for the 100 million diabetics in the known world, this can be best accomplished by educating in groups. PMID- 7494741 TI - The patients' voice: testimonies from patients suffering from chronic disease. AB - A medical congress brings together those doctors who are interested in the proposed theme of that particular congress. Within the congress 'Patient Education 2000' an unusual aspect was the presentation, in the form of posters, of the requirements of patients suffering from one of the diseases or conditions covered by the congress. On the one hand, these posters showed the impact of the illness; on the psychological aspect, within the family, within the professional and social lives as well as the financial aspect. On the other hand, the posters showed what the patients really wanted to say to the health care team. In this paper we present a synthesis illustrating certain significant examples. PMID- 7494742 TI - Patients' experiences with their disease: learning from the differences and sharing the common problems. AB - Experiences and views of patients are presented concerning psychological, professional, family life, cognitive and financial aspects of the costs of several chronic diseases: arterial hypertension, autonomous dialysis, back pain, bronchial asthma, chronic obstructive pulmonary disease, colostomy, diabetes mellitus, epilepsy, laryngectomy, Parkinson's disease. The posters expressed what patients would really like to tell their doctors. PMID- 7494743 TI - Cross-cultural validation of existing quality of life scales. AB - Health care has recently tended to take a more global perspective when considering the individual patient. In addition to a measure of functional status, the integration of the disease into the daily life of the individual, or his quality of life, is now being evaluated. This concept, while relatively recent, is the result, in part, of changes in attitudes towards diagnosis and treatment of patients, particularly those with chronic and severe diseases. Over time, what is sought is a measurement of change in the level of well-being during the evolution of an illness. In clinical trials, the different levels that exist between 2 groups may be measured. The validation of a quality of life scale requires both a qualitative and a quantitative approach with attention being paid to the methodological issues in scale development. Specific statistical techniques are used to modify evolving versions. The cross-cultural adaptation of an existing scale into another language requires a rigorous qualitative phase before the quantitative phase is begun. The end product of this approach should be an equivalent scale, rather than a literal translation. PMID- 7494744 TI - Use, misuse and abuse of questionnaires on quality of life. AB - Questionnaires quantify formal aspects of unidirectional communication. This definition indicates the major limitations of research based on such techniques: it will be unbalanced and restrictive, and therefore incomplete. The wide use of questionnaires is often unavoidable in medical and scientific research, yet they are often developed with insufficient attention to the needs that they are intended to meet. Their design may be imperfect even in relatively straightforward circumstances such as the reporting of laboratory observations, but inappropriate as well as imperfect in others, such as the study of individual quality of life. This concept and others related to it are defined, together with some major sources of error in questionnaire development and use. Steps that can be taken to improve the situation are also mentioned. PMID- 7494745 TI - PROACTA: a way to study the tendency to occur (TTO) of patient behaviours. AB - Classical decision theory formulas have been combined with more recent theories from psycho-social origins (such as Rosenstock's health belief model, Fisbein's rational action theory, Bandura's concept of self-efficacy, Triandis' concern for the assessment of habits, etc.). A software, called PROACTA, enables simulations of actual cases. Currently, it is being tested in various backgrounds. It is orientated in order to help conceive intervening strategies, especially in the area of patient education. Examples of a case study are presented. PMID- 7494747 TI - A comprehensive system for quality improvement in ambulatory care: assessing the quality of diabetes care. AB - A comprehensive state-of-the-art system for quality improvement in ambulatory care has been designed to test (1) whether patients at 8 intervention sites demonstrate improved health status and satisfaction with their care as compared to patients at 8 control sites and (2) the extent to which timely patient self reported data influences provider practice patterns. During the study pilot period, several investigators developed, tested and analyzed disease-specific questionnaires for 7 common chronic conditions. An advanced automated information system was designed to link hospital computer information and patient questionnaire data in order to provide timely communication between patients and providers about important health problems. This report briefly describes the 3 year quality of ambulatory care clinical trial and details the development and pilot testing of the disease specific questionnaire for diabetes. Reliability testing showed correlations were higher for fixed events such as foot ulcers than for subjective judgments such as satisfaction with providers. Responsiveness testing indicated that this questionnaire could measure behavior and care modifications in patients 6 months following an outpatient education course, compared to the baseline pre-education values. Diabetes severity information has been collected from self-administered questionnaires and laboratory data to assist providers in assessing the patients' likelihood of 4-year mortality. It is anticipated that the rapid identification of important patient issues will assist patients and clinicians in mutually addressing and resolving health care problems, thereby improving the quality of outpatient care. PMID- 7494746 TI - Laryngectomized patients' education and follow-up. AB - Functional and aesthetic problems due to mutilating surgical excisions and adjuvant radiotherapy prolonging the therapeutic period and increasing consequences, are common to all cervicofacial tumors. Laryngectomy is the most representative example, because it affects the aero-digestive tract, the site of major functions; breathing, phonation and swallowing. It necessitates excellent information to the patient post-operatively, and then adapted rehabilitation requiring the learning of oesophageal speech. A close follow-up is required to track loco-regional relapses, secondary neoplastic sites and metastases, cessation of alcohol and tobacco use, and to provide family, social and professional rehabilitation. There are often problems to manage with socio culturally disadvantaged classes, and physically and psychologically fragile patients, whose hope of surviving is, in general, mediocre. PMID- 7494748 TI - Benefits of a computer-assisted education program for hypertensive patients compared with standard education tools. AB - ISIS (Initiation Sanitaire Informatisee et Scenarisee), a French computer assisted hypertension and cardiovascular risk education program, was developed to provide patients at cardiovascular risk with a modern interactive educational tool combining rigorously scientific information with the aesthetic attractiveness of multimedia communication. To test the impact of this tool on patient health information retention, 158 hypertensives hospitalized for initial work-up (day hospital) or therapeutic adjustments (3 days) were randomized into control (n = 79) and ISIS (n = 79) groups. Both groups received cardiovascular education through standard means: physicians, nurses, dietitians and pamphlets. In addition, ISIS patients underwent a 30- to 60-min session on the computer with the ISIS program. Cardiovascular knowledge was tested by the same investigator administering a standardized 28-item questionnaire before and 2 months after education. Retesting was made by telephone. At the time of first assessment, all patients were aware that they would be retested. A total of 138 completed questionnaires (69 from each group) was analyzed. Overall mean cardiovascular knowledge score before education improved significantly after education. This improvement was greater in the ISIS than the control group. These results confirm the potential of computer-assisted education in hypertensives from a specialized center. PMID- 7494749 TI - Recent advances in colostomy care. AB - Recent advances in colostomy care are described. A colostomy is a surgically created opening of the large bowel through the abdominal wall. Three kinds of stoma care are presented, paying particular attention to several advances and setbacks. The special conditions of colostomy care in China are described. PMID- 7494750 TI - Advance care planning: eliciting patient preferences for life-sustaining treatment. AB - Patient autonomy is a guiding principle in medical decision-making in America. This is challenging when patients become mentally incapacitated and cannot express their preferences. Advance care planning (ACP) addresses this challenge. ACP is a deliberative and communicative process that helps people formulate and communicate preferences for future medical care in the event of mental incapacity. Advance directives are mechanisms for communicating and/or documenting ACP, and are either instructional (e.g. statement of treatment preferences in living wills) or proxy types (e.g. appointment of another person to speak on the patient's behalf). ACP discussions between patients and health care providers and patient-orientated educational ACP materials often ignore insights from 2 related activities, health promotion and human information processing. More effective ACP should occur with greater attention to the concepts of stages of change and self-efficacy, the Health Belief Model, and the necessary requisites for cognitive integration. PMID- 7494751 TI - Developing an educational book about back pain: problems encountered--possible solutions. AB - The development of an educational book about back pain is described. It is used as a part of the patient education on the Back School of the Geneva Hospital. The problems encountered and possible solutions are presented. PMID- 7494752 TI - The transforming force of laughter, with the focus on the laughing meditation. AB - Laughter is a very special phenomenon. Unfortunately, its (psycho)therapeutic value is underestimated and its transforming force is not used to its full extent. Next to spontaneous laughter, there is also a laughing meditation, a structured exercise of 15 min with 3 stages: stretching, laughing (and/or crying), silence. The laughing meditation can be used as 'adjuvans' in all kinds of therapy and, in other situations it enhances the coping with all of life's woes. This can be done alone or in any group. It facilitates the viewing of problems from another angle and it makes life lighter. Responses from participants indicate a deep relaxation, a feeling of being 'whole', a feeling of being 'unburdened', I feel my tears or I feel sad, a feeling of acceptance of the present. PMID- 7494754 TI - Patient Education 2000: take-home messages from this congress. AB - A personal review is presented about the conclusions and highlights during the Congress on Patient Education 2000 (14 June 1994, Geneva, Switzerland). Attention is paid to differences between diseases concerning patient education. Further, a summary is presented of the discussion of quality of life, over-optimism, medical information supply, purpose of patient education, the need for discipline and the psychological problems due to wrong patient education. PMID- 7494753 TI - Arterial hypertension: lessons from patient education. AB - The face of hypertension has been changing rapidly over the last few decades, from a serious disease to a cardiovascular risk factor. Patient education has been instrumental in bringing about tremendous improvements in hypertension related mortality, morbidity, life expectancy, and life quality. Patient education has evolved from an adjunct to medical therapy to an intervention in its own right. In this process patient education tasks and techniques themselves have undergone remarkable developments, driven by evolving patient needs due to medical progress. The same is true for the roles of patients and health care providers. Dealing with hypertension is a behavior change process which demands serious learning efforts from all parties involved, patients, health practitioners and health care administrators alike. This paper focuses therefore on patient education for preventive behavior change and risk factor management. Special emphasis will be placed on processes and tools for effective patient education. PMID- 7494755 TI - The role of the World Health Organization in promoting patient education with emphasis on chronic diseases. PMID- 7494756 TI - Diabetes mellitus: lessons from patient education. AB - The education of diabetic patients, proposed as an essential therapeutic tool since the early 1920s and accepted as such by official medicine only in the 1970s, has generated great enthusiasm over the last decade, with increasing concern for greater effectiveness by improved motivation of both patients and doctors. Structured education depends on the precise definition of agreed, short term objectives, whose attainment shall be verified. Educational objectives may be set at different levels: knowledge of the disease, skills required for treatment, capacity to integrate therapy in everyday life,... The most relevant objectives however are the therapeutic goals of each individual patient, i.e. most often, prevention of acute complications, near-normoglycemia to prevent late complications and foot care to prevent disabling consequences of the latter. This can only be attained through a global approach to the patient, at once medical, educational and psychological. Medical science has definitively confirmed the importance of near- normoglycemia and proposes more effective insulin regimens and new recommendations for diet and exercise. Education demands a lot from health care providers: specific training, teaching skills, good communication, supportive attitude, readiness to listen and to negotiate. Patients' motivation to learn and adhere to treatment is also greatly influenced by individual factors, both psychological and environmental, that need to be taken into account. PMID- 7494757 TI - Back pain: lessons from patient education. AB - Education for the patient with back pain is currently being discussed and reassessed. In the 1970s and 1980s, the clinical and scientific communities were convinced that patient education as a sole treatment for back pain was most beneficial. In the 1990s outcome studies, randomized controlled trials, meta analysis and best synthesis evidence have moderated this view. This brief overview summarizes important aspects put forward in recently published articles about the education of patients with non-specific, low back pain in industrialized societies. PMID- 7494759 TI - Asthma: lessons from patient education. AB - Asthma is a common condition. Recent understanding of the disease has meant that the inflammatory process in the airways is likely to be treated by the regular use of preventative therapy even when the patient is well. The evidence suggests that such treatment is often not taken. Audit suggests that poor communication and lack of information remains a common problem. There is increasing use of a team approach to the management of asthma, often necessitated by the sheer numbers of patients requiring regular review. Such an approach combined with the giving of control of their own condition to the patients, has now been shown to improve outcomes. PMID- 7494758 TI - Parkinson's disease--medical education and psychosocial aspects. AB - This article describes the attempt to develop educational material for patients suffering from Parkinson's disease (PD) and for their partners. The material relates to the neuropathology of PD. The extent of the degeneration of dopaminergic neurons in the substantia nigra, present at the outbreak of the disease, is illustrated to the patient. The substantia nigra is explained as a TV broadcasting system, the nerve fibers to the so-called striatum as cables and the striatum itself is explained as a TV-set with 2 channels (called dopamine D 1 receptor and dopamine D 2-receptor). The action of anti-Parkinsonian agents is exemplified with the substance L-DOPA, a precursor of the transmitter dopamine. The patient and his partner are introduced to the time-course of the L-DOPA concentration in blood after oral intake of the substance (the pharmacokinetics of L-DOPA). The pharmacokinetics can explain the clinical improvement and the long-term effects of L-DOPA therapy. The second part reports on psychosocial problems that Parkinsonian patients and their spouses encounter and how psychological interventions may help to solve some of these psychosocial problems. PMID- 7494760 TI - Cost-benefit analysis of patient education. AB - As the growth of healthcare continues to outpace inflation, health policy makers are increasingly concerned with cost containment. This article summarizes studies of the cost-benefit of patient education studies in managed care and other settings, including comprehensive approaches to healthcare delivery that emphasize education. On the average, for every dollar invested in patient education, $3-4 were saved. The article concludes with recommendations for healthcare policy makers and administrators. PMID- 7494761 TI - Patient education with reference to the process of behavioral change. AB - From a traditional aspect, behavioral change is conceptualized as a shift from one stable state to another, from stable unhealthy behavior to stable healthier behavior. This approach will influence the type of intervention program to be adopted as well as the type of assessment instrument applied. A concept supported by the results of studies involving a large number of subjects, which enable the natural process of behavioral change to be observed, suggests that behavioral change is a non-stable condition with distinct stages called precontemplation, contemplation, action, and maintenance. There may be relapse at any stage and this should be interpreted as a natural part of the change process. Health educators are confronted with different tasks at each level, and by addressing these tasks and tailoring the intervention programs to suit the stage of the individual subject, better support during the process of change can be given. A project involving cooperations between university institutes and the pharmaceutical industry has developed and evaluated education programs which address different stages of change in a primary care setting. Action-oriented programs for the prevention of coronary disease, sleep disturbance, and chronic pain have proven to be effective. Target behaviors are smoking, stress management, nutrition, and physical exercising. These programs have recently been complimented by others addressing precontemplation and contemplation stage subjects. Further research will have to be carried out in order to find assessment instruments designed to assign the right program to the right person. PMID- 7494764 TI - Nonsurgical treatment of multinodular nontoxic goitre. PMID- 7494762 TI - Health beliefs and knowledge of patients and doctors in clinical practice and research. AB - This paper calls for a renewal of interest in monitoring and improving patients' knowledge about their condition and self-care. Such knowledge is essential for patients with chronic disorders such as diabetes. Small or non-existent relationships often observed between knowledge levels and metabolic outcomes in diabetes can be seen to result from inappropriate investigation of these relationships. Knowledge about different aspects of diabetes management needs to be related to relevant outcomes and metabolic control cannot be expected to be related to every aspect of knowledge about diabetes self-care. The importance of health beliefs and perceived control in determining treatment choice and efficacy of treatment is considered. The implications of patients' and doctors' varying knowledge and beliefs for their preferences for different treatment options are examined and the need to consider such preferences when designing, developing and evaluating new treatments is emphasised. PMID- 7494763 TI - Helicobacter pylori and radical therapy for peptic ulcer. PMID- 7494765 TI - Porphyria and vampirism: another myth in the making. PMID- 7494766 TI - The Babinski reflex. AB - The plantar response is a reflex that involves not only the toes, but all muscles that shorten the leg. In the newborn the synergy is brisk, involving all flexor muscles of the leg; these include the toe 'extensors', which also shorten the leg on contraction and therefore are flexors in a physiological sense. As the nervous system matures and the pyramidal tract gains more control over spinal motoneurones the flexion synergy becomes less brisk, and the toe 'extensors' are no longer part of it. The toes then often go down instead of up, as a result of a segmental reflex involving the small foot muscles and the overlying skin, comparable to the abdominal reflexes. With lesions of the pyramidal system, structural or functional, this segmental, downward response of the toes disappears, the flexion synergy may become disinhibited and the extensor hallucis longus muscle is again recruited into the flexion reflex of the leg: the sign of Babinski. A true Babinski sign denotes dysfunction of the pyramidal tract, and should be clearly distinguished from upgoing toes that do not belong to the flexion synergy of the leg. Correct interpretation of the plantar response depends only to a minor degree on the method or site of stimulation of the foot. It is therefore most important to assess the response in the entire leg. PMID- 7494767 TI - Medical examinations for pilots. AB - High altitude is an environment that is alien to man. Stressors associated with the mechanics of flight include motion and acceleration forces applied in three vectors and in degrees that are foreign to the human vestibular apparatus. Furthermore, the work patterns of a pilot can interfere with lifestyle and circadian rhythms. Therefore medical fitness is an important consideration in determining an individual's suitability to exercise a pilot's licence. The medical standards applied depend on the type of aircraft flown and the duties expected of a pilot. There are three broad categories of pilot. In ascending order of stringency of medical standards these are the private pilot, the professional pilot and the military pilot. PMID- 7494768 TI - Dealing with the difficult patient. AB - Dealing with difficult patients can represent a significant burden in the life of doctors. It is more productive, however, to view this burden as a product of the interaction between doctor and patient, for which both have a responsibility, rather than attributing any problems encountered to shortcomings of the patient alone. There is a significant risk in such situations of potentially harmful over medicalisation. It behoves doctors, therefore, to try to prevent such problems becoming established, or make some attempt to rectify matters if they have already become so. Much is known about the factors that contribute to successful and unsuccessful clinical transactions. The awareness of what doctors bring both as professionals and as individual people to this interaction, will count as much as the practical clinical efforts made towards helping patients. PMID- 7494770 TI - Rehabilitation medicine. AB - The nature of rehabilitation medicine is outlined in terms of its objectives, its methods, the patient groups which it seeks to help and its relationships to other specialties. Some major advances of recent years are discussed, such as the development of the impairment/disability/handicap framework. With particular emphasis upon neurological rehabilitation, the breadth of the specialty is then illustrated with examples of current preoccupations. These include recovery patterns of the damaged nervous system, testing the efficacy of existing therapies, applying new treatment techniques and developing quantitative measures of disability and handicap. Looking to the future, some key areas are identified where further advances might be sought. PMID- 7494769 TI - Breast cancer in the elderly. AB - Breast cancer in the elderly has attracted considerable interest in recent years for three main reasons. Firstly, information concerning the profile (clinical and biological) of the disease in the geriatric population is scarce; secondly, the number of patients is increasing, and thirdly there are conflicting data regarding the actual effectiveness of the different treatments. The present review attempts to outline the specific characteristics of this malignancy in the elderly in terms of histological pattern, stage at diagnosis, and outcome. The feasibility of standard therapies (mastectomy/lumpectomy, axillary clearance, and radio-therapy) in the elderly is carefully analysed, and compared to the efficacy of less aggressive procedures. The use of tamoxifen as a primary treatment, instead of surgery, is critically reconsidered, as well as its use as an adjuvant therapy. Finally, the effectiveness of systemic therapy in advanced disease is also discussed. PMID- 7494772 TI - Unrestricted availability of a plasma paracetamol assay service resulting in an increased number of inappropriate requests. AB - Previously, prior approval from the on-call chemical pathologist was required in our hospital for plasma paracetamol measurements. However, since May 1992, there have been no restrictions on ordering this assay. We have assessed the consequences of this policy change by comparing the number and appropriateness of requests for plasma paracetamol measurements in Chinese patients admitted to our hospital with acute poisoning over two six-month periods (July-December) in 1991 and 1993. Requests were considered appropriate if paracetamol ingestion was suspected or unknown drugs were ingested. The number of patients having plasma paracetamol concentrations assayed increased from 51 in 1991 to 141 in 1993 (176%). The corresponding increase in the number of Chinese patients admitted to two of our eight general wards with poisoning was estimated to be 93%. The proportion of 'appropriate' plasma paracetamol measurements dropped from 55% in 1991 to 21% in 1993. Eight patients had plasma paracetamol concentrations above the recommended treatment line; they were all from the group in whom the requests were appropriate. Three of the 135 patients in the group with 'inappropriate' requests were found to have slightly elevated but far from toxic plasma paracetamol concentrations. Unrestricted availability of plasma paracetamol measurements resulted in an increase in the number of inappropriate requests. PMID- 7494773 TI - Acute confusion in a young man. PMID- 7494771 TI - The use of rheumatoid factors in clinical practice. AB - Rheumatoid factor is a commonly requested test in the investigation of joint problems. Although the test has been found to be both sensitive and specific for rheumatoid arthritis in patients attending Rheumatology Clinics, when the test is applied to general hospital or community populations it performs poorly. Thus the setting in which rheumatoid factor tests are requested and awareness of the test's limitations are critical for appropriate use and interpretation of results. We studied 295 consecutive requests for testing at four centres. The majority were performed for the investigation of joint problems but only 6% of these were positive, suggesting unselective requesting. Half of the results were used to make diagnoses and influence management. However, significant numbers of clinicians felt that the test result had either excluded or confirmed a diagnosis of rheumatoid arthritis suggesting that results were being over-interpreted and that clinical decisions made on this basis may have been inappropriate. We conclude that in this setting rheumatoid factor testing is of limited value and generates misleading information. PMID- 7494774 TI - Dysphagia in a diabetic patient. PMID- 7494775 TI - Wandering consolidation. PMID- 7494776 TI - Seizures in a patient with diffuse scleroderma. PMID- 7494777 TI - Anaemia in a child. PMID- 7494778 TI - Giant retroperitoneal hemangiopericytoma. AB - Hemangiopericytomas are rare vascular tumours that are derived from pericytes. Retroperitoneal hemangiopericytomas are usually bulky but clinically silent when diagnosed because of their slow rate of growth. A 49-year-old man, who presented with only vague symptoms of abdominal fullness for several months, was found on computed tomography to have a huge well-defined mass with areas of low attenuation and well-enhanced septa. The tumour was successfully resected and was confirmed to be a malignant retroperitoneal hemangiopericytoma. It measured 30 cm in the greatest dimension. We are prompted to present this case as it is believed to be the largest tumour reported to date. PMID- 7494779 TI - Carcinoid syndrome due to a malignant somatostatinoma. AB - Somatostatinoma is one of the rarest tumours of the endocrine pancreas. Cardinal manifestations of a somatostatinoma include gallstones, mild diabetes mellitus, steatorrhoea, diarrhoea and dyspepsia. Like any other pancreatic islet cell carcinoma, a somatostatinoma may also produce several different hormones such as adrenocorticotropic hormone, calcitonin, vasoactive intestinal polypeptide, pancreatic polypeptide, gastrin, insulin, and glucagon. In many cases, the clinical picture is dominated by the effect of these other hormones. We present a patient with somatostatinoma in which an immunocytochemical study of the specimens from pancreas and liver showed a weak positive reaction for gastrin besides a strong positive reaction for somatostatin. Interestingly, this patient also showed the signs of carcinoid syndrome which was successfully treated with octreotide. PMID- 7494781 TI - Side-effects of coumarin. PMID- 7494782 TI - 5-Alpha-reductase deficiency in a Saudi "girl". PMID- 7494780 TI - Primary actinomycotic lung abscess. PMID- 7494783 TI - Dementia, myopathy, and idiopathic hypoparathyroidism. PMID- 7494784 TI - Hyperglycaemia induced by paracetamol. PMID- 7494785 TI - A minibus driver with amnesia. PMID- 7494786 TI - Answering patients' questions about transfusions. PMID- 7494787 TI - Functional endoscopic sinus surgery. PMID- 7494788 TI - A positive approach to preparation. PMID- 7494789 TI - Resistance to antibiotics in general practice. PMID- 7494790 TI - Chloramphenicol eye drops: a dangerous drug? PMID- 7494791 TI - Effective management of vaginal thrush. PMID- 7494793 TI - The modern face of whooping cough. PMID- 7494792 TI - The many guises of septicaemia. PMID- 7494794 TI - A new large-scale process for taxol and related taxanes from Taxus brevifolia. AB - PURPOSE: In view of the demonstrated antitumor activity of taxol, ready availability of the drug is important. The current isolation methods starting from the bark of Taxus brevifolia involve multiple manipulations, leading to only taxol and in a yield of 0.01%. A new process consisting of a single reverse phase column is introduced here, and the present purpose is to determine its large scale applicability. METHODS: The chloroform extractable fraction of the bark of T. brevifolia is applied directly on to a C-18 bonded silica column in 25% acetonitrile/water, with elution using a step gradient: 30-50% acetonitrile/water. On standing, eight different taxanes, including taxol, crystallize out directly from different fractions. The crystals are filtered and purified further by recrystallization. Taxol and four other taxanes are purified this way. The other three require a short silica column. Taxol is freed from cephalomannine by selective ozonolysis. RESULTS: The large scale process gave taxol (0.04%), 10-deacetylbaccatin III (0.02%), 10-deacetyl taxol-7-xyloside (0.1%), 10-deacetyl taxol-C-7-xyloside (0.04%), 10-deacetyl cephalomannine-7 xyloside (0.006%), taxol-7-xyloside (0.008%), 10-deacetyl taxol (0.008%) and cephalomannine (0.004%). Processing of the needles of T. brevifolia gave brevifoliol (0.17%), and that of the wood, 10-deacetyl taxol-C-7-xyloside (0.01%) and 10-deacetyl taxol-C. CONCLUSIONS: The reverse phase column process is simpler (one column, direct crystallization), more efficient (eight taxanes obtained simultaneously) and also gives higher yields. PMID- 7494795 TI - Characterization of alkylating versus intercalating anticancer drug-induced effects on cell survival, cell cycle kinetic and morphonuclear pattern of three neoplastic cells lines growing in vitro. AB - PURPOSE: The influence of three alkylating and three intercalating anticancer drugs on cell survival, cell cycle kinetics and chromatin patterns was monitored in vitro on three neoplastic cell lines. METHODS: This monitoring was carried out by means of the digital cell image analysis of Feulgen-stained nuclei. RESULTS: Results show that in term of cytotoxicity, the intercalating drugs were more potent than the alkylating ones. As for the cell kinetics assessment, most of the experimental conditions led to a blockage of the cells in the G2 phase of the cell cycle. A study of chromatin patterns by means of digital cell image analysis enabled us to describe 15 morphonuclear parameters. The results show that the drugs tested induced specific morphonuclear modifications, e.g. an increase in nuclear size. The 15 morphonuclear parameters were submitted to multivariate analyses, i.e. principal-components analyses followed by the canonical transformation of the data. The results of these multivariate analyses enabled us to discriminate between the alkylating and the intercalating drugs. CONCLUSIONS: We conclude that it would be possible to "diagnose" the mechanism of action of DNA interacting agents (alkylating or intercalating drugs) by means of the combination of digital cell image and multivariate analysis. PMID- 7494796 TI - Distribution, metabolism and tumoricidal activity of doxorubicin administered in sorbitan monostearate (Span 60) niosomes in the mouse. AB - PURPOSE: Encapsulation of doxorubicin in niosomes was sought as a route to tumour targeting and improved tumoricidal through the alteration of doxorubicin pharmacokinetics and metabolism. METHODS: Doxorubicin niosomes (10 mg kg-1 doxorubicin) prepared from sorbitan monostearate (Span 60), cholesterol and choleth-24 (a 24 oxyethylene cholesteryl ether) in the molar ratio 45:45:10 were administered intravenously to female NMRI mice bearing the MAC 15A subcutaneously implanted tumour. Plasma doxorubicin was fractionated by gel filtration and quantified by HPLC with fluorometric detection as niosome-associated doxorubicin and released doxorubicin. Tumoricidal activity of the formulation was assessed by the intravenous injection of 5 mg kg-1 and 10 mg kg-1 doxorubicin niosomes to male NMRI mice bearing a 6 day old MAC 15A tumour. RESULTS: At least 90% of the plasma doxorubicin was associated with the niosome fraction 4 h after dosing, and 50% was still associated after 24 h. The clearance of doxorubicin released from the niosomes was about 10 fold greater than the clearance of niosomal doxorubicin (176.5 mL h-1 and 16.2 mL h-1, respectively). The area under the plasma level time curve increased 6 fold when doxorubicin was administered in niosomes, compared to doxorubicin solution (66.0 micrograms.h mL-1 and 10.3 micrograms. h mL-1, respectively). The area under the tumour level time curve was increased by over 50% by the administration of doxorubicin in niosomes when compared to the drug administered in solution (58.6 micrograms.h mL-1 and 34.3 micrograms.h mL-1, respectively). There was no statistically significant difference between levels of the drug in the heart when niosomal doxorubicin or doxorubicin solution were administered. Doxorubicin metabolites, namely doxorubicinol and the aglycones doxorubicinone, doxorubicinolone and 7-deoxydoxorubicinone, were found associated with the niosomes in the plasma, possibly due to their adsorption to the vesicle surface once formed outside the niosome. Overall metabolite levels in the liver were increased when doxorubicin niosomes were administered compared to the drug in solution. A 5 mg kg-1 injection of doxorubicin niosomes produced a terminal mean tumour weight that was similar to that obtained from animals administered 10 mg kg-1 doxorubicin solution. CONCLUSIONS: Modest tumour targeting was achieved by the delivery of doxorubicin in sorbitan monostearate niosomes, increasing the tumour to heart AUC0-24 ratio from 0.27 to 0.36 and a doubling of tumoricidal activity. The overall level of doxorubicin metabolites was also increased. PMID- 7494797 TI - Development of a flow microcalorimetry method for the assessment of surface properties of powders. AB - PURPOSE: This study describes the development of a microcalorimetric flow cell which allows powder surface energetics to be probed my means of measuring their interaction with water vapour. METHODS: A flow cell has been constructed and tested in an isothermal microcalorimeter to assess the interaction between water vapour and powder surfaces. The cell was constructed to mix two separate air streams (0% and 100% relative humidity respectively) to create any humidity at a standard flow rate. The powder sample was equilibrated in dry air and then exposed to sequential increments in humidity. RESULTS: Adsorption isotherms were constructed from the cumulative heat as a function of humidity. It was possible to differentiate between different samples of alpha-lactose monohydrate (which appeared identical by contact angle determination). It was also possible to measure adsorption to two different alkyl p-hydroxybenzoates which were hydrophobic and of low surface area. CONCLUSIONS: This technique offers a very sensitive and versatile method of obtaining a reliable indication of powder surface energetics and as such is a major advance in the field. PMID- 7494798 TI - The effect of particle fragmentation and deformation on the interparticulate bond formation process during powder compaction. AB - PURPOSE: The compression behaviour and the compactability of particles have been studied. In addition, an expression describing the bond strength over a tablet cross section was derived and these calculated values were compared with the experimentally determined tablet tensile strength values. METHODS: The compression behaviour of particles of a series of size fractions of four materials were assessed by tablet surface areas (particle fragmentation propensity) and by yield pressures (particle deformability), derived from in die Heckel profiles. The porosity and the tensile strength of the tablets were also determined. RESULTS: Sodium chloride and sodium bicarbonate possessed limited fragmentation while the converse applied for sucrose and lactose. Sodium chloride and sodium bicarbonate were the extreme materials with respect to particle deformability and compactability. Except for sodium chloride, a limited effect of original particle size on the compactability of the particles was observed. CONCLUSIONS: The observations on the compactability of the powders was explained by postulating that fragmentation affects mainly the number of bonds in a compact cross section, while deformation affects mainly the bonding force of these bonds, through a relationship with the contact area between a pair of particles. The deviations between the predicted strength of particle-particle bonds and the determined tensile strength values was explained by a high bonding capacity of some particles, e.g. due to an unpredicted high surface deformability, or by a fracture mechanic effect during tablet strength determination. PMID- 7494799 TI - Improved static compression behaviors and tablettabilities of spherically agglomerated crystals produced by the spherical crystallization technique with a two-solvent system. AB - PURPOSE: Poorly compressible crystals of acebutolol hydrochloride were agglomerated by the spherical crystallization technique with a two-solvent system to improve the compressibility for direct tabletting. The mechanism of improvements in static compression behaviors and tablettabilities of the spherically agglomerated crystals were investigated. METHODS: The improvement of static compression behaviors of the agglomerated crystals was determined by measuring the stress relaxations and elastic recoveries of compressed powder of original and agglomerated crystals. The improved tablettability of agglomerated crystals was evaluated by the pressure transmission ratio upon compression, the ejection pressure for releasing the tablet from the die and the tablet strength, i.e., tensile strength required for breaking. RESULTS: The higher relaxation pressure and the lower elastic recovery of the agglomerated crystals than of the original crystals were found. The pressure transmission ratio data showed that the friction pressures of the two crystals were similar during the compression period. The ejection pressure of the agglomerated crystals was lower than that of the original crystals. The tensile strength of the tablet of agglomerated crystals was greater than that of the original crystals. CONCLUSIONS: The compressibility and tablettability of the spherically agglomerated crystals prepared by the spherical crystallization technique were much improved due to their increased plastic property and reduced adhesive property compared to the original crystals. PMID- 7494800 TI - Compressed donut-shaped tablets with zero-order release kinetics. AB - PURPOSE: Simple uncoated compressed tablets with a central hole (donut-shape) are proposed to provide a constant drug release over a long period of time (> 20 hrs). The effect of hole size and drug solubility on the release kinetics is investigated. METHODS: The donut-shaped polyethylene oxide (PEO, Mw = 4 x 10(6)) tablets (600 mg and 12 mm diameter) are bored with a drill bit (3/32", 7/64", 1/8", and 5/32"). RESULTS: The release of theophylline from the donut-shaped tablets is zero order (80-90% release) before rapidly decreasing. As the hole size is increased from 7/64" to 5/32", the release rate increases and the release time is shortened. However, the release of theophylline from the donut-shaped tablet with a hole size of 3/32" follows the same anomalous release profile from a tablet without a hole. As drug solubility increases, the duration of linear drug release is shortened to 65-70% release followed by a severe tailing at the later stage of the release. CONCLUSIONS: Donut-shaped PEO tablets with a hole provide zero-order release kinetics because the effect of the releasing surface area on the release kinetics is reduced. PMID- 7494801 TI - Oral solid controlled release dosage forms: role of GI-mechanical destructive forces and colonic release in drug absorption under fasted and fed conditions in humans. AB - PURPOSE: This study was undertaken to examine the effects of mechanical destructive forces on drug release from controlled release (CR) dosage forms in vitro and in vivo and their colonic release, using two CR tablets of acetaminophen A and B, showing slower and faster erosion rates, respectively. METHODS: In vitro release rates were determined by several official methods. Tablets were administered to healthy volunteers under fasting and fed conditions. RESULTS: Both tablets showed similar release rates under mild destructive conditions (e.g., paddle method at 10 rpm) but CR-B showed faster release under highly destructive conditions (e.g., rotating basket method at 150 rpm), where the tablet was eroded. The in vivo release from CR-B was faster than from CR-A, possibly because of enhanced erosion. The variable in vivo release from CR-B indicated large inter-subject differences in destructive GI forces. The fastest in vivo release from CR-B among individuals was approximated by the in vitro dissolution determined by destructive methods such as the rotating basket at 150 rpm. The slowest in vivo release from tablets A and B was lower than the dissolution by the paddle method at 10 rpm. The release from both tablets was markedly reduced at 3-4 hrs after dosing irrespective of feeding conditions which can be attributed to release inhibition in the colon. CONCLUSIONS: Effects of GI destructive forces on the tablet erosion and the release inhibition in the colon must be considered in the development of CR dosage forms. PMID- 7494802 TI - Physicochemical, pharmacokinetic and pharmacodynamic evaluation of liposomal tacrolimus (FK 506) in rats. AB - PURPOSE: Tacrolimus (FK 506) is a new potent immunosuppressant. Because of poor water solubility, the conventional intravenous dosage forms of FK 506 (C-FK 506) contain surfactants such as HCO-60 which may cause adverse effects. We sought a liposomal formulation of FK 506 (L-FK 506) containing endogenous phospholipids to target drug to the spleen, a major organ controlling the immune system. METHODS: L-FK 506, consisting of 0.1 micron diameter vesicles of phosphatidylcholine and phosphatidylglycerol (molar ratio 9:1) and 7.5 mole% drug, was evaluated for in vitro stability. The intravenous disposition profile, spleen distribution, and immunosuppression of L-FK 506 was compared with that of C-FK 506 in the rat after single doses of 0.3 mg/kg. RESULTS: The L-FK 506 showed good in vitro stability. L-FK 506 exhibited an increased volume of distribution at steady-state (Vss) (from 3.41 to 14.71 L/kg) and increased mean residence time (MRT) (from 2.83 to 16.07 hr). FK 506 concentrations in spleen were increased by 40% at 10 hr after administration of the liposomal formulation. The pharmacodynamics of L-FK 506, evaluated by the extent of inhibition of splenocyte proliferation, was comparable to that of C-FK 506. CONCLUSIONS: Liposomal FK 506 may be an improved dosage form for parenteral use. PMID- 7494803 TI - Long-circulating emulsions (oil-in-water) as carriers for lipophilic drugs. AB - PURPOSE: Rapid clearance of parenterally administered oil-in-water emulsions from blood by the reticuloendothelial system (RES), mainly macrophages of the liver and spleen, has been one of the major obstacles for delivering lipophilic drugs to cells other than those in the RES. The purpose of this study therefore is to overcome this problem and develop emulsions that will have prolonged blood circulation time. METHODS: A series of amphipathic polyethylene-glycol (PEG) derivatives have been included as co-emulsifier into emulsions composed of Castor oil and phosphatidylcholine. The effect of amphipathic PEG on reducing the RES uptake and prolonging the blood circulation of the emulsion particles has been tested in vivo using mice as an animal model. RESULTS: Inclusion of PEG derivatives such as Tween-80 or dioleoyl N-(monomethoxy-polyethyleneglycol succinyl)phosphotidylethanolamine (PEG-PE) into emulsions composed of Castor oil and phosphatidylcholine decreases the RES uptake and increases blood residence time of the emulsions. The activity of PEG derivatives in prolonging the circulation time of emulsions depends on the PEG chain length (PEG2000 > PEG5000 > PEG1000, Tween-80) and the PEG density on emulsion surface. CONCLUSIONS: Inclusion of amphipathic PEG as emulsifier into oil-in-water emulsions is a very effective method to prolong the blood half life of the emulsions. Emulsions with long circulating half life in blood should be very useful as a delivery vehicle for lipophilic drugs. PMID- 7494805 TI - Adsorption of salmon calcitonin to PLGA microspheres. AB - PURPOSE: The interaction of salmon calcitonin (sCT) and poly (d,l-lactide-co glycolide) was detected during preparation and evaluation of microspheres. The purpose of this study was to quantitate the extent and nature of the interaction. METHODS: Blank microspheres were prepared by an aqueous emulsification solvent extraction technique. Adsorption studies were carried out at six concentrations of sCT and three concentrations of microspheres. Adsorption isotherms were constructed using the Langmuir and Freundlich treatments. RESULTS: Adsorption at 1 mg/ml sCT concentration resulted in almost complete depletion of the peptide from the adsorption medium with the time to reach maximum adsorption decreasing with increasing microsphere concentration. At sCT concentrations below 100 micrograms/ml, a true equilibrium occurred in 1 hour or less while at higher concentrations (up to 350 micrograms/ml), a transient equilibrium was reached in 1 to 2 hours, followed by further adsorption of the peptide. The adsorption followed the Langmuir isotherm at concentrations below 200 micrograms/ml, indicating formation of a monolayer. Multilayer interaction, described by the Freundlich isotherm, occurred at higher concentrations and resulted in complete depletion of sCT from the adsorption medium. The affinity constant during monolayer formation was 0.09 and the plateau surface concentration was 5.1 micrograms/mg. The multilayer peptide-peptide adsorption showed a lower affinity (0.025) but higher capacity (24 micrograms/mg) than the monolayer peptide-polymer adsorption. CONCLUSIONS: The results show that poly (d,l-lactide-co-glycolide) microspheres have a high adsorption capacity for sCT which must be considered in formulating a controlled delivery product of this peptide. PMID- 7494804 TI - Pharmacokinetics of iopromide liposomes in rabbits. AB - PURPOSE: The dose-proportionality of pharmacokinetics of an iodinated contrast medium, iopromide, encapsulated into liposomes was investigated. METHODS: Following single intravenous administration of 150 mg iodine/kg (potential diagnostic dose) and a five-fold higher dose in rabbits the pattern of elimination was studied until 7 d and the blood concentrations were monitored up to 72 h after administration. The iodine concentration in the liver was calculated on the basis of the blood concentration and related to the concentration measured in the rabbit liver. RESULTS: The dose-normalized blood concentration-time profiles of the encapsulated iodine were not superimposable. Contrary to the low dose a steady-state concentration of 2.8 mg iodine/mL was observed in blood for 60 min after the high dose administration indicating a saturation of the liposomal liver uptake. For both doses the elimination of iodine occurred predominantly via the kidneys and was complete 7 d after administration. The dose-normalized amounts of iodine excreted with the urine were similar for both dose groups. From the blood data it was calculated that doses up to about 300 mg iodine/kg should result in a dose-proportional increase of liposomal liver uptake before saturation occurs. This was confirmed by the measured iodine liver concentrations after increasing the doses stepwise from 150 to 750 mg iodine/kg. CONCLUSIONS: In rabbits for the dose range 150 to 750 mg iodine/kg iopromide liposomes reveal dose-dependent pharmacokinetics due to a saturation in liver uptake which occurs for doses of 300 mg iodine/kg corresponding to 300 mg lipid/kg onwards. PMID- 7494807 TI - Effect of aging on the kinetics of blood-brain barrier uptake of tryptophan in rats. AB - PURPOSE: The purpose of this investigation was to examine the effect of aging on the blood-brain barrier (BBB) transport of tryptophan. METHODS: A well established in-situ brain perfusion technique was used to examine brain uptake of 14C-tryptophan in 2-, 12- and 24-month old Sprague-Dawley rats; perfusate tryptophan concentrations ranged from 0.00175 to 2 mM. Uptake data were modeled using non-linear regression analysis. RESULTS: Permeability-surface area product (PA) for tryptophan was significantly lower in 12- and 24-month old rats, as compared to the 2-month old animals. A transport model consisting of both saturable (Michaelis-Menten type) and non-saturable components best described brain uptake of tryptophan in all 3 age groups. However, age-dependent differences in BBB transport parameters of tryptophan were observed. For the saturable component, both Vmax and Km were significantly lower in the 12- and 24 month old rats, as compared to the youngest group of rats. These results suggest that transporter mobility, number and affinity for tryptophan are altered in older rats. Values for Kd, the rate constant for non-saturable brain tryptophan transport, were also significantly lower in animals of the two older age groups. Interestingly, PA values for thiourea, a compound believed to be transporter across BBB by diffusion, were also lower in these two age groups. CONCLUSIONS: Aging decreases the ability of the BBB to transport the neutral amino acid tryptophan. PMID- 7494806 TI - Effects of insulin concentration and self-association on the partitioning of its A-21 cyclic anhydride intermediate to desamido insulin and covalent dimer. AB - PURPOSE: In the pH range 2-5, human insulin degrades via deamidation at the A-21 asn and covalent dimerization. Both products form via a common cyclic anhydride intermediate, a product of intramolecular neucleophilic attack by the A-21 carboxyl terminus. This study examines the influence of [insulin] and self association on the partitioning of the intermediate to products. METHODS: Insulin self-association was characterized (pH 2-4) by concentration difference spectroscopy. Deamination rates (pH 2-4) and concurrent rates of covalent dimer formation (pH 4) were determined versus [insulin] at 35 degrees C by initial rates. A mathematical model was developed to account for the overall rate and product composition profile versus pH and [insulin]. RESULTS: Between pH 2-4, insulin self-associates to form non-covalent dimers with a pH independent association constant of 1.8 x 10(4) M-1. The overall rate of degradation is governed by intermediate formation, while product distribution is determined by competition between water and the phe B-1 amino group of insulin for the anhydride. In dilute solutions, deamidation is first-order in [insulin] while covalent dimerization is second-order. Thus, deamidation predominates in dilute solutions but the fraction of covalent dimer formed increases with [insulin]. At high [insulin], self-association inhibits covalent dimer formation, preventing exclusive degradation via this pathway. The model accurately predicts a maximum in covalent dimer formation near pH 4. CONCLUSIONS: A mechanism is described which accounts for the complex dependence of insulin's degradation rate and product distribution profile on pH (between 2-5) and [insulin]. If these results can be generalized, they suggest that covalent aggregation in proteins may be inhibited by self-association. PMID- 7494808 TI - Uptake of fractionated 3H-heparin by isolated rat Kupffer cells. AB - PURPOSE AND METHODS: The uptake of fractionated 3H-heparin by isolated rat Kupffer cells was examined to determine the uptake mechanism. RESULTS: The association of fractionated 3H-heparin was concentration-dependent with a dissociation constant of 3.4 nM and a maximum association capacity of 1.3 pmol/10(6) cells, suggesting the involvement of a specialized mechanism. Although 2,4-dinitrophenol inhibited neither the association nor internalization of fractionated 3H-heparin, lowering the temperature from 37 degrees C to 4 degrees C reduced the internalization of fractionated 3H-heparin by 70% without affecting the association. CONCLUSIONS: It is suggested that the uptake mechanism may differ from receptor-mediated endocytosis of polypeptides and be mediated by scavenger receptors, because organic anions, and several ligands of scavenger receptors, as well as several heparin analogs, inhibit the binding of fractionated 3H-heparin to Kupffer cells, while phenylarsine oxide, which is known to inhibit the receptor-mediated or absorptive endocytosis of polypeptides, inhibits neither the association nor internalization of fractionated 3H-heparin. PMID- 7494809 TI - Pharmacokinetic/pharmacodynamic evaluation of deflazacort in comparison to methylprednisolone and prednisolone. AB - PURPOSE: The pharmacokinetics and pharmacodynamics of deflazacort after oral administration (30 mg) to healthy volunteers were determined and compared with those of 20 mg of methylprednisolone and 25 mg of prednisolone. METHODS: Methylprednisolone, prednisolone and the active metabolite of deflazacort, 21 desacetyldeflazacort, were measured in plasma using HPLC. For the assessment of pharmacodynamics, differential white blood cell counts were obtained over 24 hours. An integrated pharmacokinetic-pharmacodynamic (PK-PD) model was applied to link corticosteroid concentrations to the effect on lymphocytes and granulocytes. RESULTS: Deflazacort is an inactive prodrug which is converted rapidly to the active metabolite 21-desacetyldeflazacort. Maximum concentrations of 21 desacetyldeflazacort averaged 116 ng/ml and were observed after 1.3 h. The average area under the curve was 280 ng/ml.h, and the terminal half-life was 1.3 h. 21-Desacetyldeflazacort was cleared significantly faster than both methylprednisolone and prednisolone. The PK-PD-model was suitable to describe time course and magnitude of the observed effects. The results were consistent with reported values for glucocorticoid receptor binding affinities for the investigated compounds. CONCLUSIONS: Due to the short pharmacokinetic half-life of its active metabolite, pharmacodynamic effects of deflazacort are of shorter duration than those of methylprednisolone and prednisolone. The PK-PD model allows good prediction of pharmacodynamic effects based on pharmacokinetic and receptor binding data. PMID- 7494810 TI - pGlu-L-Dopa-Pro: a tripeptide prodrug targeting the intestinal peptide transporter for absorption and tissue enzymes for conversion. AB - PURPOSE: The purpose of this study is to investigate the characteristics of pGlu L-Dopa-Pro as a prodrug of L-Dopa. METHODS: pGlu-L-Dopa-Pro and L-Dopa-Pro were synthesized using the standard procedures of peptide synthesis. The conversion of pGlu-L-Dopa-Pro to L-Dopa was studied using pyroglutamyl aminopeptidase I and prolidase. With rats as the animal model, the stability of pGlu-L-Dopa-Pro in intestinal homogenates was determined, then the transport characteristics of pGlu L-Dopa-Pro were studied using in-situ perfusion and Ussing chambers. RESULTS: pGlu-L-Dopa-Pro, relatively stable in intestinal homogenates and intestinal fluid, had a dimensionless permeability of 1.8 at 0.04 mM. Its intestinal permeability was significantly inhibited by 20 mM captopril, by a mixture of dipeptides, 80 mM Gly-Gly and 5 mM Gly-Pro, and by 2 mM cephradine. Further, in Ussing chambers, its mucosal to serosal permeability decreased dramatically with concentration. Conversion studies showed that pGlu-L-Dopa-Pro was degraded by pyroglutamyl aminopeptidase I, an enzyme releasing the N-terminal pyroglutamic acid, with Vmax and Km of 0.6 mumole/min/g protein and 21 mM, respectively, and that L-Dopa-Pro was degraded by prolidase with Vmax and Km of 44 mumole/min/g protein and 0.48 mM, respectively. CONCLUSIONS: This tripeptide, a potential prodrug of L-Dopa, is absorbed by the intestinal peptide transporter, is relatively stable in the gut wall, and is converted to L-Dopa by peptidases with the cleavage by pyroglutamyl aminopeptidase I to L-Dopa-Pro as the rate limiting step. PMID- 7494811 TI - Differential induction of UDP-glucuronosyltransferase activity and gene expression in rat liver. PMID- 7494813 TI - Patient-oriented pharmaceutical research: focus on the individual. PMID- 7494812 TI - Hepatic membrane transport of organic cations using isolated rat hepatocyte membrane vesicles: structure-transport relationships. PMID- 7494814 TI - Pharmaceutical solids: a strategic approach to regulatory considerations. AB - PURPOSE: This review describes a conceptual approach to the characterization of pharmaceutical solids. METHODS: Four flow charts are presented: (1) polymorphs, (2) hydrates, (3) desolvated solvates, and (4) amorphous forms. RESULTS: These flow charts (decision trees) are suggested as tools to develop information on pharmaceutical solids for both scientific and regulatory purposes. CONCLUSIONS: It is hoped that this review will lead to a more direct approach to the characterization of pharmaceutical solids and ultimately to faster approval of regulatory documents containing information on pharmaceutical solids. PMID- 7494815 TI - Diffusion in HPMC gels. I. Determination of drug and water diffusivity by pulsed field-gradient spin-echo NMR. AB - PURPOSE: This work describes diffusivity measurements of drug (adinazolam mesylate) and water in a variety of solutions including polymer gels. METHODS: Pulsed-field-gradient spin-echo (PFGSE) NMR methods were employed to measure the diffusivity. RESULTS: In binary component solutions, adinazolam diffusivity is generally found to exhibit an exponential dependence on the concentration of the viscosity-inducing agent (VIA), which is glucose, lactose, maltoheptaose, hydroxypropyl methylcellulose (HPMC) or drug itself. An increasing obstruction power to drug diffusion from glucose to HPMC is observed, which can be related to the polymerization degree of the VIA. In contrast, adinazolam diffusivity in HPMC gels shows little dependence upon the polymer viscosity grades examined (K100LV, K4M, and K15M). The temperature dependence of adinazolam diffusivity in dilute VIA solutions reveals that the diffusion barrier for the drug is similar to that for self-diffusion of water. CONCLUSIONS: The retarding effect from the VIA for drug diffusion is concluded to be primarily associated with a steric obstruction mechanism. In multicomponent gels with varied concentrations of drug, lactose and HPMC, the drug diffusivity can be approximately described as an exponential function of the summation of the products of the proportionality constant (Ki) and concentration for each VIA component. In contrast, water diffusion behavior shows an universal exponential dependence upon the VIA concentration and small dependence upon the nature of the VIA. The interpretation of the diffusivity data is discussed and compared to two existing diffusion models (Yasuda and Mackie Meares models). PMID- 7494816 TI - Diffusion in HPMC gels. II. Prediction of drug release rates from hydrophilic matrix extended-release dosage forms. AB - PURPOSE: A mathematical model is described for the prediction of the relative change in drug release rate as a function of formulation composition for HPMC based extended-release (ER) tablets of adinazolam mesylate and alprazolam. METHODS: The model is based on the equation derived by Higuchi for the diffusional release of soluble drugs from polymeric matrices and on our recent measurements of the concentration dependency of adinazolam diffusivity in dilute HPMC gels and solutions. The assumptions made in applying the model include (i) that diffusion is the sole mechanism of drug release (i.e. swelling kinetics are ignored), and (ii) that the surface area-to-volume ratio and concentrations of adinazolam, lactose and HPMC in the gel layer are proportional to that of the dry tablet. RESULTS: Reasonable correlations were obtained between the experimental drug release rate ratios and the predicted drug release rate ratios for ER adinazolam mesylate (R2 = 0.82) and low-dose (0.5 mg) ER alprazolam tablets (R2 = 0.87). The predictive power for a 6-fold higher dose of ER alprazolam tablets was not as good (R2 = 0.52). CONCLUSIONS: These results are consistent with previous knowledge of the release mechanisms of these formulations. ER adinazolam mesylate and ER alprazolam 0.5 mg exhibit primarily a diffusion controlled release mechanism, while ER alprazolam 3 mg deviates from pure diffusional release. The limitations of the model are discussed and point to the need for continued study of the swelling kinetics of matrix ER systems. PMID- 7494817 TI - A new method for estimating dermal absorption from chemical exposure. 3. Compared with steady-state methods for prediction and data analysis. AB - PURPOSE: This paper compares unsteady-state and steady-state methods for estimating dermal absorption or analyzing dermal absorption data. The unsteady state method accounts for the larger absorption rates during short exposure times as well as the hydrophilic barrier which the viable epidermis presents to lipophilic chemicals. METHODS: Example calculations for dermal absorption from aqueous solutions are presented for five environmentally relevant chemicals with molecular weights between 50 and 410 and log10Kow between 0.91 and 6.8: chloromethane, chloroform, chlordane, 2,3,7,8-TCDD, and dibenz(a,h)anthracene. Also, the new method is used to evaluate experimental procedures and data analyses of in vivo and in vitro permeation measurements. RESULTS: In the five example cases, we show that the steady-state approach significantly underestimated the dermal absorption. Also, calculating permeability values from cumulative absorption data measured for exposure periods less than 18 times the stratum corneum lag time will overestimate the actual permeability. CONCLUSIONS: In general, steady-state predictions of dermal absorption will underestimate dermal absorption predictions which consider unsteady-state conditions. Permeability values calculated from data sets which include unsteady-state data will be incorrect. Strategies for analyzing in vitro diffusion cell experiments and confirming steady state are described. PMID- 7494818 TI - Evaluation of retinoid lactones as topical therapeutic agents in dermatology. AB - PURPOSE: Optimization of the therapeutic ratio of analogs of the topically active 11-cis, 13-cis-12-hydroxymethylretinoic acid, delta-lactone (1) relative to antihyperproliferation and antihyperkeratinization vs. toxicity. METHODS: Nine analogs of 1, in which variations were made in the lipophilic cyclohexenyl moiety or in the lactone ring, were evaluated for topical activity against hyperkeratinization, inhibition of TPA-induced DNA synthesis and for skin irritation. RESULTS: Although more potent lactones than the parent lactone 1 were identified, none possessed the favorable therapeutic ratio associated with 1. CONCLUSIONS: The delta-lactone 1 possesses unique molecular features responsible for its desirable therapeutic ratio as an antihyperproliferative and antihyperkeratotic agent. In view of its very low systemic retinoid toxicity and the absence of any systemic toxicity, this lactone may be a good candidate for use in the topical treatment of acne. PMID- 7494820 TI - Mechanisms involved in iontophoretic transport of angiotensin. AB - PURPOSE: The feasibility of using iontophoresis to enhance the permeation rate of a model peptide was investigated in vitro using hairless mouse skin. METHODS: Angiotensin 2 (AT 2) was employed as a permeant probe, using optimum iontophoresis conditions. A number of physicochemical parameters (donor ionic strength; valence of competitive ions; pH of donor solution) were studied with the aim of exploring the mechanisms involved in the iontophoretic transport through the skin: electrokinetic transport or convective transport. For this purpose, the magnitude of the convective solvent flow was also evaluated by the permeation of (3H) H2O. The interest of pulsed currents for peptide delivery was also investigated and the effect of current density and frequency was studied. RESULTS: AT 2 transport was found to be enhanced 20-fold in comparison to passive permeation and was found to be proportional to the current density with direct currents as with pulsed currents. CONCLUSIONS: Although the flux enhancement of ions during iontophoresis is due principally to the electrical potential gradient, secondary effects such as convective solvent flow contribute also to flux enhancement of peptide delivery. This effect is dependent of physicochemical conditions of formulation. PMID- 7494819 TI - In vitro percutaneous absorption of arildone, a highly lipophilic drug, and the apparent no-effect of the penetration enhancer Azone in excised human skin. AB - PURPOSE: Arildone, a novel lipophilic antiviral drug when evaluated in Clinical Trials showed limited skin absorption and antiviral efficacy. These studies were conducted to explain the apparent poor absorption characteristics and attempt to promote skin absorption by using Azone, a penetration enhancer. METHODS: Standard in vitro skin permeation methods using excised human skin were employed to characterise the absorption of Arildone. 14C-Arildone was used to estimate the distribution in skin layers by scintigraphic and autoradiographic procedures. RESULTS: The aqueous solubility and distribution constant values for Arildone were 2 micrograms ml-1 and 5 x 10(5) (isopropyl myristate/water), respectively. Absorption through full thickness skin or stratum corneum-viable epidermal membranes (diffusional resistant dermis removed), from a propylene glycol vehicle, was slow and the addition of Azone had no effect on the permeation rate. Distribution studies showed accumulation of Arildone in the stratum corneum. The concentration of Arildone in the viable epidermis was estimated from sectioning the skin and was found to be in sufficient amounts (400 micrograms cm-3) to have potential antiviral activity. CONCLUSIONS: The apparent accumulation of Arildone in the stratum corneum suggested that the hydrophilic skin region presented the main barrier to permeation. Azone which affected the permeability of the stratum corneum was therefore not effective at enhancing Arildone absorption. Vehicles which readily permeate and enhance the transfer of lipophilic drugs from the stratum corneum into the viable epidermis were recommended. PMID- 7494821 TI - Permeability of peptides and proteins in human cultured alveolar A549 cell monolayer. AB - PURPOSE: The transport of peptides or proteins across the alveolar cell monolayer was studied in vitro in order to elucidate their transport pathway. METHODS: The permeability of 14 peptides or proteins and 6 dextrans with MW 1,000-150,000 was measured in cultured human lung adenocarcinoma A549 cell monolayers at 37 degrees C or 4 degrees C. The stability of the tested peptides and proteins was also evaluated. RESULTS: The permeability coefficients of these macromolecules across the A549 cell monolayer at 37 degrees C ranged from 10(-5) to 10(-7) (cm/sec), and exhibited a good inverse correlation with molecular weight. All macromolecules were stable throughout the transport experiment, and degradation by proteases was minimal. Permeability at 4 degrees C did not differ from that at 37 degrees C. Clear selectivity for direction of transport was not observed. CONCLUSIONS: These results suggested that the tested peptides and proteins appeared to penetrate the A549 cell monolayer via a paracellular route by passive diffusion. PMID- 7494822 TI - Mechanisms of transport of quinapril in Caco-2 cell monolayers: comparison with cephalexin. AB - PURPOSE: To determine the transport mechanisms of quinapril and cephalexin in Caco-2 cell monolayers, a cell culture model of the human small intestinal epithelium. METHODS: Uptake, transepithelial transport and intracellular accumulations of these two drugs were measured using Caco-2 cell monolayers grown onto Millicells and magnetically stirred diffusion chambers. RESULTS: Transepithelial transport, apical (AP)4 uptake and intracellular accumulation of both drugs depended on the maintenance of a transepithelial proton gradient and temperature of the medium. However, quinapril transport and accumulation, which did not display a maximum at approximately pH 6, was more sensitive to proton gradient change, whereas cephalexin transport was more sensitive to concentration change (range 0.5-5 mM). In addition, quinapril (1 mM) transport was decreased significantly (p < 0.05) by 10 mM cephalexin, loracarbef, Gly-Pro and Phe-Pro, but not by enalapril; whereas cephalexin (0.1 mM) transport was decreased significantly (p < 0.05) by all four compounds. Similarly, AP quinapril (1 mM) uptake was also decreased by 10 mM loracarbef, Gly-Pro, cephalexin, and enalapril, but these inhibitory effects (20-50%) were quantitatively less than their inhibitory effects on cephalexin uptake (50-90%). Finally, the AP uptake of quinapril was also significantly (p < 0.05) inhibited by FCCP (10 micrograms/ml), amiloride (0.5 mM), DEP (0.5 mM), and staurosporine (5 nM). CONCLUSIONS: The transport of quinapril in the Caco-2 cells is via a combination of the carrier mediated proton gradient-dependent peptide transporter and passive diffusion. PMID- 7494823 TI - Enhanced oral bioavailability of DDI after administration of 6-Cl-ddP, an adenosine deaminase-activated prodrug, to chronically catheterized rats. AB - PURPOSE: 6-Cl-2',3'-dideoxypurine (6-Cl-ddP), an adenosine deaminase (ADA) activated prodrug of ddI, may be an effective antiretroviral agent for the treatment of AIDS dementia due to its ability to deliver increased concentrations of ddI to brain tissue. To examine the feasibility of administering this drug orally, the oral and hepatic portal bioavailabilities of 6-Cl-ddP were determined. In addition, the oral and portal bioavailabilities of ddI after administration of the prodrug were compared to those from administration of ddI itself. METHODS: Pharmacokinetic and bioavailability studies were conducted in fully conscious, chronically catheterized rats in a randomized crossover design. Plasma ddI and 6-Cl-ddP concentration-time profiles were determined by HPLC. RESULTS: 6-Cl-ddP has poor apparent oral bioavailability (7% +/- 3%, n = 3) but high bioavailability after portal administration (97% +/- 11%), suggesting either poor absorption or extensive gut wall metabolism. The appearance of > 50% of the dose as ddI in the systemic circulation after an oral dose of 6-Cl-ddP rules out poor absorption of the prodrug, and confirms expectations of high ADA activity in the gastrointestinal tract. Gastric administration of 6-Cl-ddP resulted in a > 10 fold increase in the oral bioavailability of ddI, from 3-7% to > 50%, and a significant decrease in the variability in apparent bioavailability. CONCLUSIONS: These data indicate that lipophilic adenosine deaminase activated prodrugs of dideoxypurine nucleosides may have limited utility for improving CNS delivery after oral administration but may be useful in enhancing the oral bioavailability of highly polar and therefore poorly absorbed dideoxynucleosides. PMID- 7494824 TI - Echo-planar magnetic resonance imaging to assess water volume in the distal small bowel. AB - PURPOSE: Assessment of fluid volumes and flow through the small intestine has in the past only been possible by means of invasive intubation studies on human volunteers. Intubation very likely disturbs gut motility and stimulates secretion. METHODS: The aim of this study was to utilise the new technique of echo-planar magnetic resonance imaging in order to non-invasively visualise the changing volume of water in the small intestinal lumen. 200 mls of test solution was ingested and water volume assessed using a multi-slice scanning technique on 3 separate days. The solutions were pure water, pure water plus 2.264 or 10 g of mannitol. These were taken on separate days by 8 healthy male volunteers. Regions of interest were constructed in the area of the lower pelvis excluding retroperitoneal structure. RESULTS: The water content of the lower small intestine did not change significantly over the 4 hours after the control solution. By contrast after both mannitol solutions there was an increase in the amount of water in the distal intestine as assessed by the area under the curve of the volume time profile (Control 51 ml.h (SD +/- 47); mannitol 2.264 g/200 ml 72 ml.h (SD +/- 57); 10 g/200 ml mannitol 115 ml.h (SD +/- 56)). Page's L Trend test showed that the trend for the volume to increase with increasing mannitol concentration to be statistically significant at the 1% level (L = 108). CONCLUSIONS: The study highlights the potential of echo-planar magnetic resonance imaging to visualise changes in gastrointestinal physiology in a noninvasive manner. PMID- 7494825 TI - Transport of biologically active interferon-gamma across human skin in vitro. AB - PURPOSE: Several studies have suggested epidermal uptake of cytokines, such as interferons, can be facilitated using topical liposomal formulations. We have evaluated the in vitro transport of biologically active recombinant human interferon-gamma (rhIFN-gamma) into and through split-thickness human skin to assess this possibility. METHODS: Skin samples were exposed to rhIFN-gamma under various conditions involving hydrated and dry surface conditions in the presence and absence of liposomes. A new low-level ELISA and an anti-viral bioassay were used to quantitate transported rhIFN-gamma. Immunohistochemical staining for ICAM 1 expression by keratinocytes was used to visualize the extent and distribution of rhIFN-gamma transport. RESULTS: Apparent steady-state transport of rhIFN-gamma occurred within the first 5 hours of exposure with approximately 10% of transported rhIFN-gamma demonstrating bioactivity. While the permeability of rhIFN-gamma across human skin under drying conditions was enhanced by the presence of liposomes, no augmentation of permeability was observed when the skin was kept hydrated. Liposomal formulations of rhIFN-gamma had greater transport rates than aqueous formulations when the applied formulations were allowed to dry after dosing. CONCLUSIONS: Our results demonstrate the transport of biologically active rhIFN-gamma across human skin in vitro and suggest a role for stratum corneum hydration as one possibility for the augmented cytokine transport. PMID- 7494826 TI - Different effects of absorption promoters on corneal and conjunctival penetration of ophthalmic beta-blockers. AB - PURPOSE: The purpose of this study was to investigate the improvement in corneal penetration of ophthalmic beta-blockers of various lipophilicities afforded by absorption promoters and to compare the corneal against conjunctival penetration in response to absorption promoters. METHODS: The penetration of the beta blockers, atenolol, carteolol, tilisolol, timolol, and befunolol, in the presence of absorption promoters, across the isolated corneal and conjunctival membranes of albino rabbits was measured using a two-chamber glass diffusion cell. EDTA, taurocholic acid, capric acid, and saponin were used as the absorption promoters. RESULTS: The absorption promoters significantly increased the corneal permeability of most beta-blockers, especially the hydrophilic agents. The absorption promoters also enhanced the conjunctival permeability of beta blockers, although their effect in promoting conjunctival penetration was less than that on corneal penetration. There was a differing penetration of instilled beta-blockers in the cornea and conjunctiva in response to absorption promoters. Capric acid and saponin showed significant promoting action on corneal penetration, but not on conjunctival penetration. Taurocholic acid had a significant effect on conjunctival penetration but not on corneal penetration. Saponin caused slight irritation. CONCLUSIONS: Absorption promoters can improve the ocular delivery of beta-blockers and a selective use of absorption promoter can improve the extent and pathway of drug ocular absorption. PMID- 7494827 TI - Effects of absorption enhancers on rat nasal epithelium in vivo: release of marker compounds in the nasal cavity. AB - PURPOSE: The assessment of the effects of nasal absorption enhancers on the rat nasal epithelium and membrane permeability in vivo after a single nasal dose of the enhancers. METHODS: The release of marker compounds (protein, cholesterol and acid phosphatase) from the nasal epithelium was measured using a lavage technique. The nasal membrane permeability was determined after intravenous administration of a systemic tracer (FITC-albumin). RESULTS: The effects of the absorption enhancers could be classified into four categories. The first consisted of HP beta CD (5%), DM beta CD (2%) and RAMEB (2%) and was not different from the control (physiological saline). For the second category, DM beta CD (5%), effects were significantly higher than for the control. The third category, SGC (1%), was more active than DM beta CD (5%) but less active than the last group. The fourth, most membrane damaging, category consisted of STDHF (1%), laureth-9 (1%) and LPC (1%). Administration of these three enhancers also resulted in release of acid phosphatase, indicating that severe membrane damage occurred. The release of cholesterol from nasal epithelium was largely dependent on the cholesterol solubilisation of the absorption enhancers. The amount of cholesterol released by laureth-9 and LPC was the largest. CONCLUSIONS: The results of this in vivo study are in agreement (i.e. similarity in rank order) with morphological and ciliotoxicity studies of nasal absorption enhancers, demonstrating that this in vivo model is a valuable tool to classify nasal absorption enhancers according to their effects on the rat nasal epithelium. PMID- 7494829 TI - Metabolism of dynorphin A 1-13 in human blood and plasma. AB - PURPOSE: A detailed investigation of the metabolic routes and rates of Dyn A1-13 in human blood and plasma was performed. METHODS: Human plasma was incubated at 37 degrees C with dynorphin A 1-13 (Dyn A1-13, 15-20 microM). The generated dynorphin fragments were separated by a new ion-pair chromatographic method and identified by matrix assisted laser desorption mass spectroscopy. The kinetic behavior of parent compound and metabolites was evaluated in the absence and presence of enzyme inhibitors. RESULTS: The major plasma metabolites of Dyn A1-13 were Dyn A1-12, A2-12, A4-12 and A4-8. Further metabolites were Dyn A2-13, A3-13, A3-12, A5-12, A6-12, A7-12, A1-10, A2-10, A2-8 and A3-8. At 37 degrees C, Dyn A1 13 had a half-life of less than one minute in plasma and blood. Plasma half-lives of major metabolites ranged between 0.5 and 4 min. Inter- and intra-individual differences in healthy volunteers were 30% (c.v.). Dyn A1-13 is mainly metabolized by carboxypeptidases to Dyn A1-12 (80%) and by aminopeptidases to Dyn A2-13 (15%). Dyn A1-12 and Dyn A2-13 are predominantly converted into Dyn A2-12 (67% of Dyn A1-13). Subsequent metabolic steps yield Dyn A3-12 (16%), Dyn A4-12 (37%) and Dyn A4-8 (33%). Aminopeptidases generate Dyn A2-12, A3-12, A4-12, A5 12. ACE metabolizes Dyn A1-12 (19%), A2-12 (33%), A3-12 (34%) and A4-12 (46%). Bestatin-sensitive endopeptidases (possibly endopeptidase 24.11) metabolize 30% of Dyn A2-12. Dyn A4-8 is formed via Dyn A4-12 (23% of Dyn A4-12) and Dyn A2-10 (37% of Dyn A2-10). CONCLUSIONS: The combination of enzyme inhibition experiments and noncompartmental kinetic analysis proved to be a powerful tool for the detailed evaluation of the metabolic fate of Dyn A1-13 in human blood and plasma. PMID- 7494830 TI - Transepithelial transport of insulin: I. Insulin degradation by insulin-degrading enzyme in small intestinal epithelium. AB - PURPOSE: The purpose of this study was to determine the existence of insulin degrading enzyme (EC 3.4.22.11) (IDE) in rat intestinal enterocytes. METHODS: Subcellular fractionation, biochemical characterization, immunoprecipitation, and western blots were employed. RESULTS: Insulin-degrading activity was localized in the cytosol, constituting 92% of total insulin-degrading activity. Cytosolic insulin-degrading activity had a pH optimum of 7.5, was almost completely inhibited by IDE inhibitors (N-ethylmaleimide, 1,10-phenanthroline, EDTA, p chloromercuribenzoate, bacitracin), but was not or only weakly inhibited by others (aprotinin, chymostatin, leupeptin, and diisopropyl phosphofluoridate.) Further, cytosolic insulin-degrading activity had a Km of 78 nM, sharing a similar Km value with insulin-degrading enzyme in non-purified forms. Approximately, 87 +/- 1.7% of cytosolic insulin-degrading activity was removed by the monoclonal antibody to IDE. On the SDS gel, the molecular weight of cytosolic IDE was 110 KD which is the same as that of human IDE. CONCLUSIONS: IDE is the major enzyme which degrades insulin in enterocytes. PMID- 7494828 TI - Species dependent esterase activities for hydrolysis of an anti-HIV prodrug glycovir and bioavailability of active SC-48334. AB - PURPOSE: The in vitro fate of an ester prodrug, glycovir, was studied to determine if the species differences in the bioavailability of pharmacologically active SC-48334 observed after glycovir administration and not observed after SC 48334 administration is due to species differences in ester hydrolysis rate or species differences in absorption of the prodrug itself, and to determine the site(s) of ester hydrolysis which contributes most to species differences in the bioavailability of SC-48334 if any. METHODS: Glycovir was incubated with small intestinal mucosa, liver S9 fractions, whole blood, red blood cells (RBC) and plasma of the rat, dog, monkey (cynomolgus and rhesus) and man, and glycovir concentrations were determined by HPLC. RESULTS: The relative bioavailabilities of SC-48334 after prodrug administration to the rat, dog, monkey and man were 99, 15, 42 and 37%, respectively. After SC-48334 administration, SC-48334 was rapidly and similarly well absorbed in all species. The hydrolysis rate in the small intestinal mucosa was well correlated with the relative bioavailability of SC 48334 after prodrug administration. Among different species the hydrolysis rate of glycovir in liver S9 fractions, blood, RBC and plasma did not parallel those in the mucosa of the small intestine. CONCLUSIONS: The species differences in bioavailability of SC-48334 with the prodrug were due to species differences in hydrolysis rates of the prodrug in small intestinal mucosa. The monkey was a good animal model for prediction of esterase activity in human small intestine and relative bioavailability in man. PMID- 7494831 TI - Preparation and characterization of dextran magnetite-incorporated thermosensitive liposomes: an on-line flow system for quantifying magnetic responsiveness. AB - PURPOSE: Dextran magnetite (DM)-incorporated thermosensitive liposomes, namely thermosensitive magnetoliposomes (TMs), were prepared and characterized in order to investigate their possibility for magnetic drug targeting. METHODS: TMs containing calcein were prepared at various DM concentrations by reverse-phase evaporation of dipalmitoylphosphatidylcholine (DPPC). They were evaluated for their physicochemical properties including size, DM capture, magnetite distribution within liposomes, and temperature-dependent calcein release. Moreover, a novel on-line flow apparatus with a sample injector, a coil of tubing placed in an electromagnet, and a fluorescence detector was developed for quantifying the magnetic responsiveness of TMs. This device allowed us a real time measurement of percentage holding of TMs by magnetic field. RESULTS: Due to water-soluble property of DM, higher contents of magnetite up to 490 mg per mmol DPPC were successfully incorporated into the liposomes with DM than with conventional magnetite (Fe3O4). Thermosensitivity and lipid integrity of TMs were not influenced by inclusion of DM. Using the on-line flow system, percentage holding of TMs by magnetic field was shown to vary with several factors; it increases as the magnetic field strength increases, the fluid flow rate decreases, the magnetite content increases, and the liposome concentration increases. Typically, at 490 mg incorporated magnetite per mmol DPPC, 0.5 ml/min fluid flow rate, and high magnetic field strength (> or = 10 kiloGauss), approximately 100% of TMs were found to be held. CONCLUSIONS: The TMs were suggested to be useful in future cancer treatment by magnetic targeting combined with drug release in response to hyperthermia. PMID- 7494832 TI - Influence of nonionic liposomal composition on topical delivery of peptide drugs into pilosebaceous units: an in vivo study using the hamster ear model. AB - PURPOSE: The purpose of this study was to test the hypothesis that nonionic liposomes facilitate the topical delivery of peptide drugs into pilosebaceous units. METHODS: The hamster ear was used as a model for human pilosebaceous units. The deposition of a hydrophilic protein, alpha-interferon (alpha-IFN), into pilosebaceous units and other strata of the hamster ear 12 hours after topical in vivo application of three nonionic liposomal formulations, one composed of glyceryl dilaurate/cholesterol/polyoxyethylene-10-stearyl ether (Non 1), the second composed of glyceryl distearate/cholesterol/polyoxyethylene-10 stearyl ether (Non-2) and the third composed of polyoxyethylene-10-stearyl ether/cholesterol (Non-3), a phospholipid-based liposomal formulation (PC) and an aqueous control solution (AQ) was determined. We also determined the deposition of a hydrophobic peptide, cyclosporin-A (CsA), into pilosebaceous units and other strata of the hamster ear after topical in vivo application of these liposomal formulations and a hydroalcoholic control solution (HA). RESULTS: The deposition of alpha-IFN into the pilosebaceous units was in the order: Non-1 >> PC > Non-2 > Non-3 = AQ. The deposition of CsA into the pilosebaceous units was in the order: Non-1 >> HA > PC > Non-2 = Non-3. CONCLUSIONS: Despite differences in the hydrophobicities and size of the drug molecules, deposition into the various ear strata was significantly enhanced by the Non-1 liposomal system. PMID- 7494833 TI - Liposomal formulations of cyclosporin A: influence of lipid type and dose on pharmacokinetics. AB - PURPOSE: Liposomal formulations of Cyclosporin A (CyA)3 have been described in more than 30 publications to substitute Cremophor EL (CrEL), a triricinoleate ester of ethoxylated glycerol, as drug carrier. However, conflicting reports did not allow to draw consistent conclusions about the influence of liposomes on CyA pharmacokinetics (PK) and pharmacodynamics. METHODS: A series of liposomal CyA formulations with varying liposome composition and lipid dose but constant CyA dose was compared in rats. Data were analysed with a PK-model taking into account the varying volume of distribution with the varying lipid concentration in blood. RESULTS: Surface properties and lipid type of liposomes are not important PK predictors of liposomal CyA, at least for small dosages of liposomes. Rather, the absolute lipid amount and the lipophilicity of cyclosporins are critical factors influencing the PK of liposomal CyA. The higher the concentration of lipid in blood and the greater the lipophilicity of cyclosporin is, the higher are the concentrations of CyA in blood. CONCLUSIONS: These relations may explain the inconsistent literature results. Together with earlier observations from our group the above findings indicate, that CyA is not caged in the liposomal membranes. Reports in literature, which claim lower clearance and a lower volume of distribution of CyA in obese rats compared to lean rats, support our assumption about the involved mechanisms. A semi-quantitative model of CyA distribution is presented, which points to the variable free fraction of CyA in plasma as the crucial factor for all previously reported phenomena in liposomal CyA formulations. PMID- 7494834 TI - Quantitative structure activity relationships of catechol derivatives on nerve growth factor secretion in L-M cells. AB - PURPOSE: Although many catechol derivatives are potent stimulators of Nerve Growth Factor synthesis in L-M cells, not much is known about their mechanism of action. In order to obtain a Quantitative Structure Activity Relationship (QSAR), AM1 quantum mechanical calculations were performed on a group of 23 catechol derivatives with different levels of activity. METHODS: A set of 18 parameters/descriptors were obtained by AM1 quantum mechanical calculations for each catechol derivative. Linear combinations of the calculated descriptors were fitted to the activity (as extracted from literature data) of the compounds by using simple or multiple regression analysis. RESULTS: Good correlation with activity was obtained for specific parameters such as the adiabatic ionization potential and other 'oxidation'-related descriptors of the molecules while poor correlation was observed for most of the other parameters as, for example, for log P. CONCLUSIONS: Our results show that activity is associated with parameters related to the oxidation of the catechol derivatives, strongly supporting recent literature suggesting that an oxidative process is involved in their action. PMID- 7494835 TI - The effect of electrostatic charge interactions on release rates of gentamicin from collagen matrices. AB - PURPOSE: This work studied the effect of changes in the magnitude of electrostatic charge interactions on the release kinetics of gentamicin from collagen matrices. METHODS: The charge distribution on collagen was altered by specific charge chemistries to yield net negative charges which exhibited binding interactions with positively charged gentamicin. The adsorption isotherms were measured to characterize binding interactions and release of gentamicin from modified matrices were measured. The release rates were compared to a mathematical model based on an instantaneous desorption coupled with diffusion mechanism. RESULTS: Ninety percent of the gentamicin loaded was released from native collagen matrices in 2.5 days (one-sided slab geometry in-vitro). Succinylated collagen matrices released 70% in 2.5 days and phosphonylated collagen matrices released 50% in 2.5 days. Excellent agreement between model predictions and experiment results were obtained. CONCLUSIONS: Modified collagen can be much more effective in antibiotic therapy in sustaining release rates compared to native collagen for charged antibiotics like gentamicin. PMID- 7494836 TI - Polymeric microspheres prepared by spraying into compressed carbon dioxide. AB - PURPOSE: The objective was to prepare polymeric microparticles by atomizing organic polymer solutions into a spray chamber containing compressed CO2 (PCA process) and to study the influence of various process parameters on their morphological characteristics. METHOD: The swelling of various pharmaceutically acceptable polymers [ethyl cellulose, poly(methyl methacrylate), poly(epsilon caprolactone), poly(dl-lactide), poly(l-lactide) and poly(dl-lactide-glycolide) copolymers] in CO2 was investigated in order to find polymers which did not agglomerate during the spraying process. Poly(l-lactide) (L-PLA) microparticles were prepared by spraying the organic polymer solution into CO2 in a specially designed spraying apparatus. The effect of various process (pressure and temperature of the CO2 phase, flow rate) and formulation (polymer concentration) variables on the morphology and particle size of L-PLA-microparticles was investigated. RESULTS: Polymers with low glass transition temperatures agglomerated even at low temperatures. The formation of microparticles was favored at moderate temperatures, low polymer concentrations, high pressures and high flow rates of CO2. High polymer concentrations and low flow rates resulted in the formation of polymeric fibers. Colloidal L-PLA particles could also be prepared with this technique in a surfactant-free environment. Initial studies on the microencapsulation of drugs resulted in low encapsulation efficiencies. CONCLUSIONS: The PCA method is a promising technique for the preparation of drug containing microparticles. Potential advantages of this method include the flexibility of preparing microparticles of different size and morphology, the elimination of surfactants, the minimization of residual organic solvents, low to moderate processing temperatures and the potential for scale-up. PMID- 7494837 TI - Measurements of size distribution and density of a pharmaceutical fat emulsion, using field-programmed sedimentation field-flow fractionation (SdFFF). AB - PURPOSE: The main goal was to establish that sedimentation field-flow fractionation (SdFFF), operated with power based field programming, is effective in the characterization of a commercial emulsion, Medialipide. This emulsion is used clinically for total parenteral nutrition and it is consisted of a mixture of long-chain triglycerides (LCT, soybean oil) with medium-chain triglycerides (MCT) emulsified by phospholipids. METHODS: Different field programming methods were used in the analysis to establish the limits of applicability of the technique. RESULTS: Identical size distribution profiles were obtained under various conditions of the analysis. The density of the droplets was determined by collecting fractions from the SdFFF eluting bands, and analyzing them by photon correlation spectroscopy. The value of density of the oil droplets was changed in the SdFFF data, until best agreement with the PCS values was achieved. The value of density corresponding to the best agreement was considered as the oil density, and it was closed to the weighted average value between soybean and MCT oils. CONCLUSIONS: Field programming extends the capabilities of sedimentation field flow fractionation in handling and characterizing complex and delicate samples as Medialipide. PMID- 7494838 TI - MicroPharm-K, a microcomputer interactive program for the analysis and simulation of pharmacokinetic processes. AB - PURPOSE: The microcomputer program, MicroPharm-K (MP-K) was developed for pharmacokinetic modeling, including analysis of experimental data and estimation of relevant parameters, and simulation. The intention was to provide a user friendly, interactive, event-driven program for PC computers. METHODS: The data are ascribed to a predefined model from a library including various routes of administration, oral or intra-venous, bolus or infusion, and various compartmental interpretations, 1 to 3. Single and multiple administrations are supported. The program provides initial estimates of the parameters in most cases, and the parameters are then fitted to the model by non linear model fitting using either the Simplex, Evol, Gauss-Newton, Levenberg-Marquardt or Fletcher-Powell algorithms. The non linear model fitting is based on the maximum likelihood method, and the criterion to minimize is either the weighted least squares (Chi 2 criterion) or the extended least squares. Graphical representations of non-fitted or curve-fitted data are immediately available (including log-scale representation), as well as pharmacokinetic typical parameters such as area under the curve, clearance, volumes, time-rate constants, transfer rate constants, etc. RESULTS: Simulated and experimental data were analysed and the results were similar to those obtained by other programs. CONCLUSIONS: This non linear fitting program has been proved in our laboratory to be a very effective package for pharmacokinetic studies, including estimation and simulation. Because it is easy-to-use and runs on basic computers, the program could also be used for educational purposes. PMID- 7494839 TI - Solubilisation of drugs in micellar systems studied by eluent gel permeation chromatography. AB - PURPOSE: The purpose of the study was to investigate the potential of a chromatographic method which is based on elution gel chromatography (EGPC) in the study of solubilisation of drugs in micellar solutions. METHODS: The EGPC method differs from conventional GPC in the use of a solution of the associating surfactant as eluent (rather than solvent) and the injection of a small volume of solution of different concentration (or alternatively injection of solvent alone) to probe the association equilibrium in the eluent. The technique was applied to a study of the solubilisation of selected drugs in aqueous micellar solutions of a triblock copolymer (Synperonic-PE F127) composed of oxyethylene [E, OCH2CH2] and oxypropylene [P, OCH2CH(CH3)] units with nominal molecular formula E98P67E98. RESULTS: EGPC curves were obtained showing vacancy peaks at the elution volumes of the drugs, clearly demonstrating their solubilisation. In addition, the micelle-molecule equilibrium of the copolymer surfactant could be monitored at all times. Quantitative determination of the partition of solute between micelles and solvent phase was not possible due to the incomplete conversion of molecules to micelles in solutions of the selected copolymer. CONCLUSIONS: The EGPC technique provides evidence for the solubilisation of the drugs in aqueous solutions of Synperonic F127; a more thorough assessment of its potential for quantitative measurement of solubilisation requires the use of a surfactant which is wholly (or at least mainly) in the micellar state under the conditions of use. PMID- 7494840 TI - Unbound total (plasma) clearance approach in interspecies pharmacokinetics correlation: theophylline-cimetidine interaction. PMID- 7494842 TI - [Magnetic resonance imaging for lumbar disk pathology. incidence of false negatives]. AB - Magnetic resonance imaging (MRI) has had an impressive impact on evaluation of degenerative diseases of the spine. Nevertheless, false negatives can occur on images involving lumbar discs. Degenerative disc diseases documented on discrography and/or pathology examination of the discs can go unrecognized. Likewise sensitivity for the detection of protruding discal hernias is not totally satisfactory (20% false negatives). Finally, a magnetic resonance image visualizing displacement of the disc is not specific (10 to 15% false positives); images showing protrusion or hernia can be seen in 30% of asymptomatic patients. Although MRI gives slightly more information than other imaging techniques, false images do exist. Moreover, the usefulness of MRI to demonstrate disc disease in case of a negative CT-scan remains to be demonstrated. PMID- 7494841 TI - Synthesis and pharmacological evaluation of spiro-analogues of 5-benzyl-5-ethyl barbituric acid. PMID- 7494843 TI - [Neoplastic ureteral obstruction: drainage by percutaneous nephrostomy or double J catheterization]. AB - OBJECTIVES: Neoplasic obstruction of the ureter is observed in patients with pelvic or extra-pelvic cancer. Indications of upper urinary tract drainage need to be clarified. METHODS: Percutaneous nephrostomy or double J catheter drainage was performed in 104 patients with ureteral obstruction due to pelvic cancer (bladder, prostate, uterus, ovary, rectum) or extra-pelvic cancer (usually breast and digestive cancers). Percutaneous nephrostomy or double J catheter was the first drainage procedure in 85 and 76 patients respectively. RESULTS: Success rate in first intention drainage was 74.5% with the double J catheter. When this procedure was impossible, percutaneous nephrostomy was always performed. In 52 cases we attempted to convert the nephrostomy to double J catheter drainage. Success rate was 68.8%. The main complications observed were ineffective drainage (12.5%) and infection (10.5%). CONCLUSION: These patients have a mean survival of 6.8 months despite the chemotherapy which is facilitated by the improved renal function. PMID- 7494844 TI - [Exclusive chemotherapy in T1-T3N0 cancers of the vocal cord with complete clinical response after neoadjuvant treatment with a cisplatin-fluorouracil combination]. AB - OBJECTIVES: Evaluation of cisplatin-fluorouracil exclusive chemotherapy for invasive squamous cell carcinoma of the glottis staged as T1-T3N0 with a complete response after cisplatin-fluorouracil neo-adjuvant chemotherapy. METHODS: A retrospective analysis of the files of 69 patients with a well-differentiated untreated invasive squamous cell carcinoma, staged as T1-T3N0 with a complete response after cisplatin-fluorouracil neo-adjuvant chemotherapy is presented. Actuarial analysis (Kaplan Meier method) of survival and local failure is presented among the group of 25 patients treated with exclusive chemotherapy and the group of 44 patients in whom the local treatment (partial laryngeal surgery or radiation therapy) initially planned was maintained. RESULTS: Three-year survival and local control estimate was 91.8% and 69.3%, respectively after exclusive chemotherapy and 92.5% and 97.2% if the local treatment was performed as initially planned. Patients with local recurrence after exclusive chemotherapy were always salvaged with partial laryngeal surgery or radiation therapy resulting in an overall 100% local control and laryngeal preservation rate. CONCLUSION: Exclusive chemotherapy for T1-T3N0 glottic carcinomas with a complete response after cisplatin-fluorouracil neo-adjuvant chemotherapy should be considered especially in patients in whom preservation of voice is of utmost importance. PMID- 7494846 TI - [Myocardial infarction by spontaneous dissection of coronary arteries in a subject with type IV Ehlers-Danlos syndrome]. AB - Acute myocardial infarction with spontaneous coronary dissection was fatal in a 33-years-old man. Autopsy and family history revealed type IV Ehlers-Danlos syndrome. In this disease, conjunctive tissue is fragilized due to a deficit in type III collagen which leads to spontaneous ruptures in large and medium sized arteries. The present case is the first with a proven rupture of the coronary arteries. This disease should be entertained in young people with no atherogenous risk factor and an acute coronary disorder since peripheral skin and joint syndromes may be discreet or missing. Treatment is difficult in case of spontaneous coronary dissection. Thrombolysis is not possible and diagnosis may require angiography which is in itself dangerous due to the fragility of the coronary vessels. PMID- 7494845 TI - [Community acquired pneumonias in the region of Montpellier. Increase of pneumococci with reduced sensitivity to penicillins]. AB - OBJECTIVES: Bacteriological data indicate that there is an increased incidence of Streptococcus pneumoniae strains with an intermediate sensitivity to penicillin. The goals of the present study was i) to investigate the profile of sensitivity of Streptococcus pneumoniae isolated from patients with bacterial pneumonia in the area of Montpellier and ii) to compare this profile with the findings of the national center registry to better appreciate geographical specificity. METHODS: Fifty-six patients with bacterial pneumonia were enrolled into the study. From September 1989 to March 1994, we performed bacterial sampling including blood cultures, protected brushes and bronchoalveolar lavage specimens. We examined the antibiotic sensitivity of the germs which were isolated. All patients were followed using clinical and radiological criteria. RESULTS: A precise bacteriological diagnosis was established in 83.6% of the population. Streptococcus pneumoniae was found in 47.2% of the samples. In 19%, the strains displayed an intermediate sensitivity to penicillin. All patients recovered. CONCLUSION: We found a higher rate of resistance to penicillin in Montpellier than the common rate of the national reference center in France. The location of Montpellier closed to the Spanish border might, at least in part, explain this difference. PMID- 7494847 TI - [Fitz-Hugh and Curtis syndrome]. AB - The Fitz-Hugh and Curtis syndrome is a perihepatitis following a genital infection. It usually occurs in young women but male cases have been reported. Chlamydia trachomatis is the most frequent causal agent observed in Europe and the United States. This germ is the number one cause of salpingitis in developed countries. Clinical signs include acute or recurrent pain in the right hypochondria. Liver tests are not modified and the sonographic examination is normal. The diagnosis can be suspected on the basis of serology results using either indirect immunofluorescence with live antigens specific for serotype D or K or ELISA. Positive predictive value is 80-90% and negative predictive value 96 99%. Diagnosis is formally established with laparoscopy. In our experience with more than 100 patients, the perihepatitis (glissonitis with pseudo-membranes) is frequently associated with adherences. The peritoneum is inflammatory and there are usually a small amount of liquid. Specimens are taken for culture. Polymerase chain reaction will improve the detection of Chlamydia since this germ is very difficult to culture. Antibiotic treatment with adapted drugs given for sufficient time is effective. PMID- 7494848 TI - [Varicella in pregnancy]. AB - Chickenpox rarely occurs during pregnancy but affected patients risk not only varicella pneumonia but also fetal contamination with potentially malformative effects or severe neonatal infection depending on whether the infection occurs early or late during pregnancy. More than 15% of the affected women have detectable lesions on the chest X-ray. Respiratory distress is the main risk with mortality reaching nearly 20%. Fetal contamination occurs via transplacental transmission. Fetal malformations are observed in less than 5% of the cases when fetal infection occurs early (before the end of the fifth month) but are generally quite severe. The mechanism is apparently fetal zona a few weeks after initial infection. Antenatal diagnosis is generally obtained on the basis of sonographic findings, and identification of viral genome using polymerase chain reaction on cordocentesis or amniotic fluid biopsy samples. Screening attempts to identify fetal anomalies and evaluate fetal prognosis. Induced abortion should be discussed in cases where both fetal malformation and fetal infection are confirmed. After five months, the risk of malformation appears to be much lower. It is known however that fetal varicella can be observed if the maternal infection occurs just before delivery. The most severe forms are seen when the maternal eruption occurs during the 4 days prior to delivery. Neonatal mortality in these cases reaches 20%. Unlike varicella, there is apparently little or no risk either for the mother or for the child in case of zona during pregnancy. PMID- 7494849 TI - [Clinical and biological features of exanthematous typhus]. PMID- 7494850 TI - [Polyalgetic syndrome of the upper limbs following intensive and prolonged use of a computer keyboard]. PMID- 7494851 TI - [Conjugal circulating anticoagulant: apropos of a married couple]. PMID- 7494852 TI - [Cyclosporosis: a new entity]. PMID- 7494853 TI - [Angiosarcoma after treatment of cancer of the breast]. PMID- 7494854 TI - [Erythrocyte superoxide dismutase activity measured in normal and high risk pregnancy, as well as in full-term and premature neonates]. PMID- 7494855 TI - Molecular pharmacological aspects of histamine receptors. AB - In this article, we review the recent developments in the field of histamine research. Besides the description of pharmacological tools for the H1, H2 and H3 receptor, specific attention is paid to both the molecular aspects of the receptor proteins, including the recent cloning of the receptor genes, and their respective signal transduction mechanisms. PMID- 7494856 TI - Glucose transporter gene expression: regulation of transcription and mRNA stability. AB - The facilitated diffusion of D-glucose across the plasma membrane is carried out by a set of stereospecific transport proteins known as the glucose transporters. These integral membrane proteins are members of a gene family where tissue specific expression of one or more members will determine in part the net rate of glucose entry into the cell. The regulation of glucose transporter gene expression is a critical feature of cellular homeostasis, as defects in specific transporter expression can lead to profound alterations in cellular physiology. In this review, we provide a brief descriptive background on the family of glucose transporters and examine in depth the regulation of the two transporters expressed in adipose tissue, GLUTI, a basal growth-related transporter and GLUT4, the insulin-responsive glucose transporter. PMID- 7494857 TI - Interferon-alpha hybrids. AB - The alpha-interferons (IFN-alpha) belong to a family of polypeptides comprising several subtypes. Using recombinant DNA technology, it has been possible to create IFN hybrids that provide novel combinations of the amino acid residues from the parental protein sequences. They have been used to study structure activity relationships of IFN-alpha and interactions with the IFN-alpha receptor, and to create analogs of natural IFNs with novel properties for potential therapeutic application. The biological data obtained with these hybrids are now evaluated in terms of the published structural and homology models of IFN-beta and -alpha. PMID- 7494858 TI - Aspects of calcium-activated chloride currents: a neuronal perspective. AB - Ca(2+)-activated Cl- channels are expressed in a variety of cell types, including central and peripheral neurones. These channels are activated by a rise in intracellular Ca2+ close to the cell membrane. This can be evoked by cellular events such as Ca2+ entry through voltage- and ligandgated channels or release of Ca2+ from intracellular stores. Additionally, these Ca(2+)-activated Cl currents (ICl(Ca)) can be activated by raising intracellular Ca2+ through artificial experimental procedures such as intracellular photorelease of Ca2+ from "caged" photolabile compounds (e.g. DM-nitrophen) or by treating cells with Ca2+ ionophores. The potential changes that result from activation of Ca(2+)-activated Cl- channels are dependent on resting membrane potential and the equilibrium potential for Cl-. Ca2+ entry during a single action potential is sufficient to produce substantial after potentials, suggesting that the activity of these Cl- channels can have profound effects on cell excitability. The whole cell ICl(Ca) can be identified by sensitivity to increased Ca2+ buffering capacity of the cell, anion substitution studies and reversal potential measurements, as well as by the actions of Cl- channel blockers. In cultured sensory neurones, there is evidence that the ICl(Ca) deactivates as Ca2+ is buffered or removed from the intracellular environment. To date, there is no evidence in mammalian neurones to suggest these Ca(2+)-sensitive Cl- channels undergo a process of inactivation. Therefore, ICl(Ca) can be used as a physiological index of intracellular Ca2+ close to the cell membrane. The ICl(Ca) has been shown to be activated or prolonged as a result of metabolic stress, as well as by drugs that disturb intracellular Ca2+ homeostatic mechanisms or release Ca2+ from intracellular stores. In addition to sensitivity to classic Cl- channel blockers such as niflumic acid, derivatives of stilbene (4,4'diisothiocyanostilbene-2,2' disulphonic acid, 4-acetamido-4'-isothiocyanostilbene-2,2'-disulphonic acid) and benzoic acid (5-nitro 2-(3-phenylpropylamino) benzoic acid), ICl(Ca) are also sensitive to polyamine spider toxins and some of their analogues, particularly those containing the amino acid residue arginine. The physiological role of Ca(2+)-activated Cl- channels in neurones remains to be fully determined. The wide distribution of these channels in the nervous system, and their capacity to underlie a variety of events such as sustained or transient depolarization or hyperpolarizations in response to changes in intracellular Ca2+ and variations in intracellular Cl- concentration, suggest the roles may be subtle, but important. PMID- 7494859 TI - Circadian rhythms and cancer chemotherapy. AB - Experimental and clinical pertinent data regarding circadian rhythmicities are reviewed in (1) target tissues, i.e., healthy tissues (actively proliferating or not) and tumor tissues, (2) pharmacology of anticancer agents, (3) toxicity and tolerance of these agents and (4) antitumor activity of cancer chemotherapy. The basic concepts of chronotolerance and/or of chronoefficacy have brought new hopes for further amelioration in the management of cancer patients. Quite recent published Phase II and III clinical trials have demonstrated that new improvements on the therapeutic index have been achieved through ambulatory chronotherapy for various solid tumors. PMID- 7494860 TI - Inhibition of mitochondrial beta-oxidation as a mechanism of hepatotoxicity. AB - Severe and prolonged impairment of mitochondrial beta-oxidation leads to microvesicular steatosis, and, in severe forms, to liver failure, coma and death. Impairment of mitochondrial beta-oxidation may be either genetic or acquired, and different causes may add their effects to inhibit beta-oxidation severely and trigger the syndrome. Drugs and some endogenous compounds can sequester coenzyme A and/or inhibit mitochondrial beta-oxidation enzymes (aspirin, valproic acid, tetracyclines, several 2-arylpropionate anti-inflammatory drugs, amineptine and tianeptine); they may inhibit both mitochondrial beta-oxidation and oxidative phosphorylation (endogenous bile acids, amiodarone, perhexiline and diethylaminoethoxyhexestrol), or they may impair mitochondrial DNA transcription (interferon-alpha), or decrease mitochondrial DNA replication (dideoxynucleoside analogues), while other compounds (ethanol, female sex hormones) act through a combination of different mechanisms. Any investigational molecule should be screened for such effects. PMID- 7494861 TI - Renal handling of drugs and amino acids after impairment of kidney or liver function--influences of maturity and protective treatment. AB - Renal tubular cells are involved both in secretion and in reabsorption processes within the kidney. Normally, most xenobiotics are secreted into the urine at the basolateral membrane of the tubular cell, whereas amino acids are reabsorbed quantitatively at the luminal side. Under different pathological or experimental circumstances, these transport steps may be changed, e.g., they may be reduced by renal impairment (reduction of kidney mass, renal ischemia, administration of nephrotoxins) or they may be enhanced after stimulation of transport carriers. Furthermore, a distinct interrelationship exists between excretory functions of the kidney and the liver. That means liver injury can influence renal transport systems also (hepato-renal syndrome). In this review, the following aspects were included: based upon general information concerning different transport pathways for xenobiotics and amino acids within kidney cells and upon a brief characterization of methods for testing impairment of kidney function, the maturation of renal transport and its stimulation are described. Similarities and differences between the postnatal development of kidney function and the increase of renal transport capacity after suitable stimulatory treatment by, for example, various hormones or xenobiotics are reviewed. Especially, renal transport in acute renal failure is described for individuals of different ages. Depending upon the maturity of kidney function, age differences in susceptibility to kidney injury occur: if energy-requiring processes are involved in the transport of the respective substance, then adults, in general, are more susceptible to renal failure than young individuals, because in immature organisms, anaerobic energy production predominates within the kidney. On the other hand, adult animals can better compensate for the loss of renal tissue (partial nephrectomy). With respect to stimulation of renal transport capacity after repeated pretreatment with suitable substances, age differences also exist: most stimulatory schedules are more effective in young, developing individuals than in mature animals. Therefore, the consequences of the stimulation of renal transport can be different in animals of different ages and are discussed in detail. Furthermore, the extent of stimulation is different for the transporters located at the basolateral and at the luminal membranes: obviously the tubular secretion at the contraluminal membrane can be stimulated more effectively than reabsorption processes at the luminal side. PMID- 7494862 TI - P450 inhibitors of use in medical treatment: focus on mechanisms of action. AB - A number of cytochrome P450s are targets for compounds that are clinically used or under clinical evaluation for treatment of patients with mycotic infections, such as dermatophytosis, superficial and systemic candidiasis, cryptococcosis and aspergillosis, with skin diseases, such as psoriasis or ichthyosis, and other retinoid-sensitive malignancies, e.g., neuro-ectodermal glioma. Some of the P450 inhibitors are candidates for the treatment of hirsutism or prostate cancer, others are potent inhibitors of the P450 isomerase involved in the synthesis of thromboxane A2, a potent platelet aggregation inducer and vasoconstrictor. PMID- 7494863 TI - Mammalian deoxyribonucleoside kinases. AB - The mammalian deoxyribonucleoside kinases are deoxycytidine kinase, thymidine kinase 1 and 2 and deoxyguanosine kinase. These enzymes phosphorylate deoxyribonucleosides and thereby provide an alternative to de novo synthesis of DNA precursors. Their activities are essential for the activation of several chemotherapeutically important nucleoside analogues. In recent years, these enzymes have been thoroughly characterised with regard to structure, substrate specificity and patterns of expression. In this review, these results are reviewed and furthermore, the physiologic metabolic role of the anabolic enzymes is discussed in relation to catabolic pathways. The significance of this information for the development of therapeutic protocols and choice of animal model systems is discussed. Finally, alternative pathways for nucleoside analogue phosphorylation are surveyed, such as the phosphotransfer capacity of 5' nucleotidase. PMID- 7494864 TI - GABAB receptors. AB - GABAB receptors are a distinct subclass of receptors for the major inhibitory transmitter 4-aminobutanoic acid (GABA) that mediate depression of synaptic transmission and contribute to the inhibition controlling neuronal excitability. The development of specific agonists and antagonists for these receptors has led to a better understanding of their physiology and pharmacology, highlighting their diverse coupling to different intracellular effectors through Gi/G(o) proteins. This review emphasises our current knowledge of the neurophysiology and neurochemistry of GABAB receptors, including their heterogeneity, as well as the therapeutic potential of drugs acting at these sites. PMID- 7494865 TI - Modulation of gene expression and endocrine response pathways by 2,3,7,8 tetrachlorodibenzo-p-dioxin and related compounds. AB - The aryl hydrocarbon (Ah) receptor binds several different structural classes of chemicals, including halogenated aromatics, typified by 2,3,7,8 tetrachlorodibenzo-p-dioxin (TCDD), polynuclear aromatic and heteropolynuclear aromatic hydrocarbons. TCDD induces expression of several genes including CYP1A1, and molecular biology studies show that the Ah receptor acts as a nuclear ligand induced transcription factor that interacts with xenobiotic or dioxin responsive elements located in 5'-flanking regions of responsive genes. TCDD also elicits diverse toxic effects, modulates endocrine pathways and inhibits a broad spectrum of estrogen (17 beta-estradiol)-induced responses in rodents and human breast cancer cell lines. Molecular biology studies show that TCDD inhibited 17 beta estradiol-induced cathepsin D gene expression by targeted interaction of the nuclear Ah receptor with imperfect dioxin responsive elements strategically located within the estrogen receptor-Sp1 enhancer sequence of this gene. PMID- 7494866 TI - Engineering of human cholinesterases explains and predicts diverse consequences of administration of various drugs and poisons. AB - The acetylcholine hydrolyzing enzyme, acetylcholinesterase, primarily functions in nerve conduction, yet it appears in several guises, due to tissue-specific expression, alternative mRNA splicing and variable aggregation modes. The closely related enzyme, butyrylcholinesterase, most likely serves as a scavenger of toxins to protect acetylcholine binding proteins. One or both of the cholinesterases probably also plays a non-catalytic role(s) as a surface element on cells to direct intercellular interactions. The two enzymes are subject to inhibition by a wide variety of synthetic (e.g., organophosphorus and carbamate insecticides) and natural (e.g., glycoalkaloids) anticholinesterases that can compromise these functions. Butyrylcholinesterase may function, as well, to degrade several drugs of interest, notably aspirin, cocaine and cocaine-like local anesthetics. The widespread occurrence of butyrylcholinesterase mutants with modified activity further complicates this picture, in ways that are only now being dissected through the use of site-directed mutagenesis and heterologous expression of recombinant cholinesterases. PMID- 7494867 TI - Electron paramagnetic resonance investigation of X-irradiated poly(U), poly(A) and poly(A):poly(U): influence of hydration, packing and conformation on radical yield at 4 K. AB - Powders and films of variably hydrated poly(U), poly(A) and poly(A):poly(U) were X-irradiated at 4 K. Spectra and free radical yields were acquired at 4 K using Q band EPR spectroscopy. Evidence for electron transfer from the hydration layer to the RNA bases, supporting in part the damage transfer hypothesis of Gregoli et al. (Radiat. Res. 89, 238-254, 1985), is presented. Based on measurements of radical yield as a function of hydration, we propose that intermolecular packing and polymer conformation are dominant factors in determining free radical trapping ability in these polymers. Our annealing results indicate that increasing hydration facilitates intercluster combination reactions. PMID- 7494868 TI - Free radical formation in single crystals of 9-methyladenine X-irradiated at 10 K. An electron paramagnetic resonance and electron nuclear double resonance study. AB - Single crystals of 9-methyladenine were X-irradiated at 10 K and at 65 K and were studied using K-band EPR, ENDOR and field-swept ENDOR (FSE) techniques in the temperature range 10 K to 290 K. Three major radicals are stabilized in 9 methyladenine at 10 K. These are: MA1, the adenine anion, probably protonated at N3; MA2, the species formed by net hydrogen abstraction from the 9-methyl group; and MA3, the radical formed by net hydrogen addition to C8 of the adenine moiety. Radical MA1 decayed at about 80 K, possibly into the C2 H adduct (MA4). The other two species (MA2, MA3) were stable at room temperature. A fifth radical species was clearly present in the EPR spectra at 10 K but was not detectable by ENDOR. This species, which decayed above 200 K (possibly into MA3), remains unidentified. The radical population at room temperature is as described by previous authors. The mechanisms for radical formation in 9-methyladenine are discussed in light of the hydrogen bonding scheme and molecular stacking interactions. PMID- 7494870 TI - Defects in the kinetics of the repair of DNA double-strand breaks and inhibition of DNA synthesis in the ataxia telangiectasia AT5BI-VA cell line: comparison to a corrected hybrid, atxbc. AB - The nature of the primary biochemical defect in the human radiosensitive and cancer-prone syndrome, ataxia telangiectasia (AT), has remained obscure despite many efforts to elucidate it. In this study, AT complementation group D cells and a nearly isogenic corrected AT-hamster hybrid derivative have been analyzed for induction and repair of initial double-strand breaks (DSBs) after exposure to ionizing radiation, using a sensitive field-inversion electrophoresis technique. Results suggesting that initial levels of damage are the same in these two cell types, but indicating differences in the fast component of DNA repair, have been compared and correlated with those resulting from a study of the radioresistant DNA synthesis defect and its correction in the same cell lines. These measurements show that the radioresistant phenotype of the substantially corrected AT-hamster hybrid correlates with its higher level of fast-component DSB repair and higher level of inhibition of DNA synthesis, but that the initial damage induction does not contribute to the phenotype. We propose that the AT gene product(s) is likely to act early in a signaling pathway which controls both DNA repair and progression of cells through the phases of the cell cycle in response to ionizing radiation. PMID- 7494869 TI - Detection of hypoxic fractions in murine tumors by comet assay: comparison with other techniques. AB - The alkaline comet assay was used to detect the hypoxic fractions of murine tumors. A total of four tumor types were tested using needle aspiration biopsies taken immediately after a radiation dose of 15 Gy. Initial studies confirmed that the normalized tail moment, a parameter reflecting single-strand DNA breaks induced by the radiation, was linearly related to radiation dose. Further, it was shown that for a mixed population (1:1) of cells irradiated under air-breathing or hypoxic conditions, the histogram of normal tail moment values obtained from analyzing 400 cells in the population had a double peak which, when fitted with two Gaussian distributions, gave a good estimate of the proportion of the two subpopulations. For the four tumor types, the means of the calculated hypoxic fractions from four or five individual tumors were 0.15 +/- 0.04 for B16F1, 0.08 +/- 0.04 for KHT-LP1, 0.17 +/- 0.04 for RIF-1 and 0.14 +/- 0.01 for SCCVII. Analysis of variance showed that the hypoxic fraction in KHT-LP1 tumors is significantly lower than those of the other three tumors (P = 0.026) but that there is no significant difference in hypoxic fraction between B16F1, RIF-1 and SCCVII tumors (P = 0.574). Results from multiple samples taken from each of five RIF-1 tumors showed that the intertumor heterogeneity of hypoxic fractions was greater than that within the same tumor. The mean hypoxic fraction obtained using the comet assay for the four tumor types was compared with the hypoxic fraction determined by the clonogenic assay, or median pO2 values, or [3H]misonidazole binding in the same tumor types.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7494871 TI - Effect of bromodeoxyuridine on radiation-induced DNA damage and repair based on DNA fragment size using pulsed-field gel electrophoresis. AB - We have used biphasic linear ramping pulsed-field gel electrophoresis (PFGE) to understand the effect of incorporation of bromodeoxyuridine (BrdUrd) on radiation induced DNA damage and repair. This technique permits a determination of the fragment size distribution produced immediately after irradiation as well as during the repair period. We found that incorporation of BrdUrd increased the induction and decreased the repair of radiation damage. The fragment size distribution was consistent with a random breakage model. When we found that significantly more damage was detected after irradiation of deproteinized DNA compared to intact cells, we studied the effects of BrdUrd incorporation on the radiation response of cells or DNA at various phases of preparation for electrophoresis: cells adherent to the culture dish (A), trypsinized cells (B), agarose-embedded cells (C) and deproteinized DNA (D). Although there was a general tendency to detect more damage when irradiation was performed later in the preparation process, steps B and C were the only successive steps which were significantly different. These findings demonstrate that incorporation of BrdUrd randomly increases the induction of radiation damage and decreases its repair at the level of 200 kbp to 5 Mbp fragments. Furthermore, they confirm that the amount of damage detected depends upon the conditions of the cells or DNA at the time of irradiation. PMID- 7494872 TI - Radiation-induced DNA damage and repair in cells of a radiosensitive human malignant glioma cell line. AB - The induction and repair of DNA double-strand breaks were studied in cells of two isogenic human malignant glioma cell lines which vary in their SF2 values by a factor of approximately 30. M059J cells are radiosensitive (SF2 = 0.02) and lack the p350 component of DNA-dependent protein kinase (DNA-PK); M059K cells are radioresistant (SF2 = 0.64) and express normal levels of DNA-PK. Zero integrated field gel electrophoresis and alkaline sucrose gradient experiments indicated that equivalent numbers of DNA lesions were produced by ionizing radiation in M059J and M059K cells. To compare the capacity of both lines to repair sublethal damage, the split-dose recovery experiment after exposure to equitoxic doses of radiation was carried out. Significant sublethal damage repair was shown for M059K cells, with a 5.8-fold increase in relative survival peaking at 4 h, whereas M059J cells showed little repair activity. Electrophoresis studies indicated that more double-strand breaks were repaired by 30 min in M059K cells than in M059J cells. These results suggest that deficient DNA repair processes may be a major determinant of radiosensitivity in M059J cells. PMID- 7494873 TI - The genetic basis of cellular recovery from radiation damage: response of the radiosensitive irs lines to low-dose-rate irradiation. AB - Recovery from the lethal effects of irradiation is commonly found when cultured mammalian cells are irradiated at low dose rates when compared to the same cells irradiated at higher dose rates. However, this cellular recovery process is severely reduced or absent in a number of radiosensitive cell lines, including those derived from the human disorder ataxia telangiectasia (AT). The genetic and molecular basis of such recovery processes is not understood, despite their importance. The responses of cells of three further radiosensitive lines, irs1, irs2 and irs3, shown previously to be mutated in different genes, to low-dose rate radiation are now presented. Plateau-phase cultures of cells of the irs2 line were found to have little or no cellular recovery, while irs1 and irs3 had considerable recovery potential. In comparing the known properties of the radiosensitive lines lacking cellular recovery, including xrs, XR-1 and scid as well as AT and irs2, it is argued that the gene products lacking in these lines normally act coordinately in a specific damage-recognition pathway. The recovery pathway is likely to be associated with the rejoining of DNA double-strand breaks, since several of these recovery-defective lines have a measurable deficiency in break repair. PMID- 7494874 TI - Neutron and gamma-radiation sensitivity of plasmid DNA of varying superhelical density. AB - Several families of negatively supercoiled topoisomers of plasmid pIBI30 were prepared by a modification of the procedure of Singleton and Wells (Anal. Biochem. 122, 253-257, 1982). The average superhelical density (sigma) was determined by two-dimensional agarose gel electrophoresis and varied from -0.010 to -0.067, corresponding to a change in the number of supercoils from 3 to 19 and an effective volume change from 1.6 x 10(8) to 4 x 10(8) A3. Samples were exposed to either fission-neutron or 60Co gamma radiation and assayed for single-strand breaks by agarose gel electrophoresis. Form I DNA for all topoisomers decreased exponentially with increasing dose. The D37 values for both neutron and gamma radiation increased monotonically with increasing magnitude of sigma. Using a branched plectonemic (interwound) form for DNA over the range of sigma studied and standard (single-hit) target theory, a quantitative linear fit to (D37)-1 as a function of the effective DNA radius, S(A), was obtained. The model predicts that both the slope (a) and the intercept (b) of (D37)-1 as a function of S(A) are directly proportional to the length of DNA and the radiation fluence. Furthermore, the ratio b/a (= ro) at sigma = 0 depends only on the ionic strength of the medium and is independent of the radiation source parameters. Our results support the model and we calculate ro = 13.4 +/- 1.4 nm, a value consistent with other investigations. Our results are consistent with studies using 137Cs (Milligan et al., Radiat.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7494876 TI - Differential expression of collagen types I and III in consequential and primary fibrosis in irradiated mouse colon. AB - These studies were undertaken to understand further the pathogenesis of consequential and primary fibrosis in mouse colon after irradiation. The distal 2.5 cm of colon of C3Hf/Kam mice was irradiated with either a single dose of 27 Gy or a split dose of 2 x 14.75 Gy separated by 10 days to induce a consequential or primary fibrotic lesion, respectively. The amount of total collagen in the two lesions was quantified by hydroxyproline, and tensile strength, an assay of tissue rigidity, was measured as a function of dose and time after irradiation. The relative distribution of collagen types I, III and IV in the colon was visualized by immunohistochemistry. Collagen types I, III and IV were quantified by immunoblot techniques, and in situ hybridization was used to identify and score the cells producing procollagen mRNA types I and III as a function of time after irradiation. The hydroxyproline and tensile strength measurements demonstrated that both lesions contained significantly increased amounts of collagen compared to controls. However, the ulcerated lesion of consequential fibrosis contained three times as much collagen and required a three- to fourfold increase in the peak force to rupture the colon as did the non-ulcerative lesion of primary fibrosis. The fibrosis accompanying the consequential lesion contained elevated levels of both collagen types I and III, but primary fibrosis contained only elevated levels of type I collagen compared to controls. The in situ hybridization studies showed cells producing increased amounts of procollagen mRNA 8 and 25 weeks before the elevated levels of collagen were detected for consequential and primary fibrosis, respectively. The cells producing the excess collagen mRNA were identified as fibroblasts. No distinction between the two lesions could be made based on the cell types producing the collagen. The distribution of labeled cells was localized to tissue areas showing specific immunofluorescence for the collagen types. These data show that the two histologically distinct lesions in irradiated colon, consequential and primary fibrosis, contain different collagen types and quantities of each type, suggesting that the underlying pathogenesis of these two lesions may be different. PMID- 7494875 TI - Comparative assessment of single-dose and fractionated boron neutron capture therapy. AB - The effects of fractionating boron neutron capture therapy (BNCT) were evaluated in the intracerebral rat 9L gliosarcoma and rat spinal cord models using the Brookhaven Medical Research Reactor (BMRR) thermal neutron beam. The amino acid analog p-boronophenylalanine (BPA) was administered prior to each exposure to the thermal neutron beam. The total physical absorbed dose to the tumor during BNCT using BPA was 91% high-linear energy transfer (LET) radiation. Two tumor doses of 5.1 Gy spaced 48 h apart (n = 14) or three tumor doses of 5.2 Gy, each separated by 48 h (n = 10), produced 43 and 70% long-term (> 1 year) survivors, respectively [corrected]. The outcome of neither the two nor the three fractions of radiation was statistically different from that of the corresponding single fraction group. In the rat spinal cord, the ED50 for radiation myelopathy (as indicated by limb paralysis within 7 months) after exposure to the thermal beam alone was 13.6 +/- 0.4 Gy. Dividing the beam-only irradiation into two or four consecutive daily fractions increased the ED50 to 14.7 +/- 0.2 Gy and 15.5 +/- 0.4 Gy, respectively. Thermal neutron irradiation in the presence of BPA resulted in an ED50 for myelopathy of 13.8 +/- 0.6 Gy after a single fraction and 14.9 +/- 0.9 Gy after two fractions. An increase in the number of fractions to four resulted in an ED50 of 14.3 +/- 0.6 Gy. The total physical absorbed dose to the blood in the vasculature of the spinal cord during BNCT using BPA was 80% high LET radiation.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7494877 TI - Errors in exposure assessment, statistical power and the interpretation of residential radon studies. AB - To date, epidemiological studies of risk from residential radon have not convincingly demonstrated an association with lung cancer. These case-control studies, however, have inherent limitations due to errors in estimates of exposure to indoor radon. These errors take on special significance because the level of residential risk predicted from studies of underground miners is relatively low and possibly at the limit detectable by current epidemiological methods. To illustrate the problem caused by errors in exposure assessment, a series of case-control studies were simulated and resulting dose-response relationships evaluated. For each of four assumed error distributions for exposure to radon progeny, 10 indoor radon studies of 700 cases and 700 controls were generated randomly from a population with a risk of radon-induced lung cancer based on extrapolations from studies of underground miners. When exposures were assumed as known without error, 6 of 10 studies failed to find a significant dose response, in accord with the theoretical power of the study of 0.47. For simulations in which exposures were measured with error, the situation was worse, as the power of the study was reduced further and it was even less likely that a single study would result in a significant finding. For each error scenario, combining data from the 10 simulated studies did result in a significant dose response. However, the pooled results are somewhat misleading, because the effects of mobility, missing radon measurements, residential occupancy and potential confounding variables such as cigarette smoking were not taken into account. Empirical estimates of power were computed using 1,000 simulated case control studies. When mobility and missing radon measurements in prior homes were incorporated into the design, the power of the study decreased, reducing the chance of detecting a significant effect of exposure. Enlarging study size to 2,000 cases and 2,000 controls increased the power of the study to 0.90 when exposure error was absent and subjects lived in one home only, but power was below 0.40 under realistic conditions for exposure error and mobility. When studies were generated under an assumption that exposure does not increase risk, up to 15% of simulated studies with 700 cases and 700 controls resulted in an estimated dose-response parameter in excess of the dose response from studies of miners. With increasing mobility and exposure error, it became virtually impossible to distinguish between the distributions of risk estimates from simulated studies based on an underlying excess relative risk of 0.015/working level month from estimates based on no risk from exposure.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 7494879 TI - Effects of low doses and low dose rates of external ionizing radiation. PMID- 7494878 TI - Differential sensitivity of two predominant stromal progenitor cell subpopulations in bone marrow to single and fractionated radiation doses. AB - The sensitivity of fibroblastoid precursor cells in rat bone marrow to single and fractionated doses of gamma rays delivered in vivo was measured. In vitro colonies were classified as being compact or diffuse, and the progenitor cells for both types were slowly cycling in vivo (survival levels after exposure to hydroxyurea were 90 +/- 6% and 93 +/- 11%, respectively). The progenitor cells forming diffuse colonies were more resistant (D0 = 1.39 Gy) than those forming compact colonies (D0 = 0.76 Gy). The fractionation sensitivities were characterized by an alpha/beta ratio of 12.7 +/- 5.5 Gy for diffuse colonies and 4.5 +/- 3.0 Gy for compact colonies, respectively. The progenitor cells forming diffuse colonies may contribute more to long-term regeneration after high doses in vivo. PMID- 7494880 TI - Osteoporosis as a model for the long-term clinical consequences of the menopause. AB - The decrease in ovarian function culminating in the last menstrual period, the menopause, in women in their forties and fifties takes place over a period of 3 to 5 years. There are several immediate clinical consequences of the menopause that are easy to detect and treat, usually with hormone replacement. In contrast, the longer term consequences of the menopause, cardiovascular disease and osteoporosis, are usually clinically silent during these early postmenopausal years. As a result, it has proven difficult to get physicians and their patients to accept this cause-and-effect relationship between the menopause and the development of osteoporosis or heart disease a decade or more later. The details concerning the menopause and osteoporosis are by now so well established that this disease can serve as a model for studies linking the menopause to heart disease. This article, with its focus on osteoporosis, serves as an introduction to the later articles detailing the cardiovascular consequences of the menopause. PMID- 7494881 TI - The nonhuman primate model of the relationship between gonadal steroids and coronary heart disease. AB - Experimental data derived from studies using cynomolgus macaque females provide strong evidence that estrogen influences both premenopausal and postmenopausal coronary artery atherosclerosis. Because the monkey studies are not hampered by selection bias, the data are supportive of the tentative conclusions from epidemiological studies indicating that the association between postmenopausal estrogen use and reduced coronary artery atherosclerosis is real. Present forms of HRT are not sufficiently acceptable to women to result in a compliance rate likely to affect the adverse impact of estrogen deprivation on coronary artery atherosclerosis. Nutritional supplementation may provide a more acceptable alternative. PMID- 7494882 TI - The epidemiology of coronary heart disease and estrogen replacement in postmenopausal women. AB - Cardiovascular diseases remain the leading cause of death in American women. Evidence from epidemiological studies indicates that estrogen replacement therapy in postmenopausal women can reduce their risk of coronary heart disease. Current users, in particular, appear to enjoy the most substantial cardiac benefits. It remains possible that the protective effect of estrogen observed in these studies may be due, in part, to bias. Women who use estrogen see a physician regularly, and compliance with estrogen therapy may identify a low-risk group of women. Furthermore, women who choose to use estrogen may lead generally healthier lifestyles than those who do not take such medication. However, adjustment for known cardiac risk factors in many of the large studies had little impact on the results, implying an equivalent risk status for users and nonusers. In addition, both animal and human studies show that estrogen use lowers low-density lipoprotein cholesterol levels, increases high-density lipoprotein cholesterol levels, improves blood flow, and reduces atherosclerosis. The effect of progestin added to estrogen therapy has not been adequately assessed, but initial evidence suggests that it may somewhat attenuate, although not completely eliminate, the benefits of estrogen on cardiovascular disease. Finally, each individual woman differs in her particular risks and benefits, and careful consideration of each woman's unique situation is required; the fear of breast cancer should be considered, together with the knowledge that many interventions are available to reduce the risk of cardiovascular disease. PMID- 7494883 TI - Coronary arteriography in estrogen-treated postmenopausal women. AB - Four cross-sectional studies that used coronary arteriography to determine the extent of coronary atherosclerosis have provided some of the most convincing evidence that estrogen replacement reduces cardiovascular risk in postmenopausal women. One study of outcome in women undergoing coronary arteriography at baseline has found that the greatest improvement in total mortality occurred in women with significant coronary stenosis. Angiographic studies have also confirmed the importance of high-density lipoprotein in mediating the effect of estrogen and the beneficial effect of estrogen on endothelial function. PMID- 7494884 TI - Estrogen replacement therapy and stroke. AB - The results of the 19 studies examining the association between estrogen replacement therapy and cerebrovascular disease are inconsistent. Although all 7 studies of death from stroke found a 20% to 60% reduction in risk among estrogen users relative to nonusers, studies of incident stroke and subarachnoid hemorrhage in particular are conflicting, with RRs from 0 to 2.3. Although a protective effect of estrogen on stroke is biologically plausible, information regarding effects of dose, duration, and discontinuation of estrogen is limited and contradictory. Additionally, selection bias, recall bias, and confounding cannot be completely discounted. Further studies are needed to determine estrogen's biological effects; the effects of varying doses, durations, and types of estrogen; the consequences of combined estrogen/progestogen regimens and of other routes of administration; and whether other factors (such as age and smoking) modify or confound estrogen's effect on stroke risk. PMID- 7494885 TI - Practical aspects of hormone replacement therapy. AB - Menopause is an endocrinopathy characterized by hypoestrogenism. Estrogen can be replaced easily. Available preparations can be taken by mouth, intramuscularly, transdermally, or transvaginally. Unopposed estrogens increase the risk of endometrial cancer, which can be overcome by the concomitant administration of a progestin agent. The progestin can be administered cyclically either every month, resulting in a monthly menses, or every 3 months, resulting in menses every 3 months. The progestin can also be administered every day, which achieves amenorrhea in over half of the patients; the remainder have breakthrough bleeding. Abnormal bleeding can be assessed by either endometrial biopsy or transvaginal ultrasound. PMID- 7494886 TI - Androgen and progestagen effects on plasma lipids. AB - Androgens are 19-carbon steroid rings. Progestagens include both 19-carbon and 21 carbon steroid rings; the 19-carbon progestagens are generally more androgenic than are the 21-carbon compounds. Both androgens and progestagens are physiological regulators of plasma lipids, particularly high-density lipoprotein (HDL) cholesterol. The structure of a particular hormonal preparation, as well as its route of administration, modulates its regulatory effects. Both endogenous and exogenous androgens have a suppressive effect on HDL cholesterol in males, with little effect on other plasma lipoproteins. Oral and nonaromatizable androgens have a greater suppressive effect on HDL cholesterol, particularly on HDL2, than do aromatizable androgens. Cross-sectional studies in males generally show a positive relationship between serum T and plasma HDL levels; data in females suggest an inverse relationship between androgens and HDL cholesterol. Medroxy-progesterone acetate and related progestagens have a mild suppressive effect on plasma HDL levels. The C-19 compounds have a greater suppressive effect on HDL cholesterol and the HDL2 density subfraction. PMID- 7494887 TI - QNU. The union at work. PMID- 7494888 TI - A big issue taking shape: who or what will shape the response? PMID- 7494890 TI - Can enterprise bargaining work in the aged care area? PMID- 7494889 TI - Get to know the nurses of the future. PMID- 7494891 TI - Protective assistance guidelines. PMID- 7494892 TI - Toxic shock syndrome. PMID- 7494894 TI - Political interviews. 1. Prime Minister Paul Keating. PMID- 7494893 TI - Issues in occupational health and safety. Workers' compensation and changes to common law. PMID- 7494895 TI - Political interviews. 2. Leader of the Opposition John Howard. PMID- 7494896 TI - Anonymity in peer review--time for a change? PMID- 7494897 TI - Reconciling science and policy in setting federal drinking water standards--four states' perspectives. AB - After almost 20 years of experience with implementing the Safe Drinking Water Act and eight with Amendments to the Act, the individual states within the United States have gained valuable experience while trying to reconcile the legal mandates provided by the statutes with the science underlying them. This paper presents four different topics illustrating the problems of reconciling these two issues in the regulation of toxic chemicals in drinking waters. It presents these from the perspectives of the states of Massachusetts, Rhode Island, Connecticut, and New Jersey and offers suggestions for improved program efficiency based on considerations of comparative human health risks. The approach and schedule for controlling toxic chemicals used through 1994 are first examined and a recommendation is made for more flexibility in the rate at which chemicals are regulated. Recent U.S. EPA proposals to more stringently control radon in drinking waters are presented in the context of all sources of radon exposures, illustrating the intersection of science, laws, and economic consequences of regulatory initiatives. Inhalation and dermal exposures as a result of using chemically contaminated drinking waters are then discussed with the suggestion of the possible underprotectiveness of some present standards. Finally, the difficulty faced by the states and federal government in the control of naturally occurring arsenic exposures through drinking water is also presented and an argument is made for more local flexibility in the application of health-based standards. PMID- 7494898 TI - A case study of cancer data set combinations for PCBs. AB - Results of several animal bioassays have demonstrated the carcinogenic potential of polychlorinated biphenyl (PCB) mixtures. Although PCBs are no longer manufactured, cancer risk assessment for PCBs remains an important issue because of continued potential human exposure from many sources. The existing cancer risk estimate for PCBs used by the U.S. EPA is based on liver tumors observed in female Sprague-Dawley rats in a lifetime bioassay. Liver cancer has been observed in other long-term bioassays as well. In this case study, experimental designs and biological characteristics of the data from these studies were evaluated to determine whether a combination of the data sets is scientifically reasonable. A statistical analysis of the data sets based on likelihood ratio theory was used to assess the compatibility of individual data sets to a common multistage dose response model. The results from these biological and statistical assessments suggest that at least two data sets could be combined to derive a quantitative risk estimate for PCBs. Increased confidence in the quantitative estimate would result from such combination because more data are being used to assess the dose response relationship. PMID- 7494899 TI - Neurotoxicity modeling for risk assessment. AB - The setting of acceptable exposure levels for neurotoxicants has followed the traditional approach of dividing experimental no-observed-adverse-effect-levels (NOAELs) by safety/uncertainty factors. NOAELs are believed by many toxicologists to represent levels having zero or negligible risk, while uncertainty factors are used to account for a number of sources of variation. Although the use of NOAELs in this manner has been criticized because of their imprecise quantitative definition, NOAELs for nonquantal neurotoxic effects have not been replaced by more precisely defined quantities (e.g., benchmark doses), partly due to the absence of a generally accepted methodology for attaching specific risk levels to low exposures. The present paper describes a quantitative approach to modeling nonquantal neurotoxic effects for risk assessment, which can be used to obtain results similar to the familiar results obtained in risk assessment for carcinogenicity and developmental toxicity. The steps involved in implementing the process are discussed, with particular attention being given to the critical step of defining an adverse neurologic effect. An experimental data set is used to illustrate the methodology. PMID- 7494901 TI - Navigating the narrows: industry's role in risk management. PMID- 7494900 TI - A physiologically based pharmacokinetic model for fluoride uptake by bone. AB - A sex-specific, physiologically based pharmacokinetic (pbpk) model has been developed to describe the absorption, distribution, and elimination of fluorides in rats and humans. Growth curves generated by plotting mean body weights (kg) against age (weeks or years) are included in the simulation model to allow the integration of chronic fluoride exposure from birth to old age. The model incorporates age and body weight dependence of the physiological processes that control the uptake of fluoride by bone and the elimination of fluoride by the kidneys. Six compartments make up the model. These are lung, liver, kidney, bone, and slowly and rapidly perfused compartments. The model also includes two bone subcompartments: a small, flow-limited, rapidly exchangeable surface bone compartment and a bulk virtually nonexchangeable inner bone compartment. The inner bone compartment contains nearly all of the whole body content of fluoride, which, in the longer time frame, may be mobilized through the process of bone modeling and remodeling. The model has been validated by comparing the model predictions with experimental data gathered in rats and humans after drinking water and dietary ingestion of fluoride. This physiological model description of absorption, distribution, and elimination of fluoride from the body permits the analysis of the combined effect of ingesting and inhaling fluorides on the target organ, bone. Estimates of fluoride concentrations in bone are calculated and related to chronic fluoride toxicity. The model is thus useful for predicting some of the long-term metabolic features and tissue concentrations of fluoride that may be of value in understanding positive or negative effects of fluoride on human health. In addition, the pbpk model provides a basis for across-species extrapolation of the effective fluoride dose at the target tissue, bone, in the assessment of risk from different exposure conditions. PMID- 7494902 TI - Development and submission of a nonclinical (pharmacology/toxicology) CANDA. AB - The experience with the submission of a nonclinical (pharmacology and toxicology) computer-assisted New Drug Application (CANDA) is reviewed. This system consisted of a stand-alone personal computer running several commercial programs in Microsoft Windows to access both text and data. WordPerfect was used as the word processor that contained all the documents and data tables (in read-only format) that were submitted in hard copy, and Andyne GQL was used as a tool to query the data in an Oracle relational database. Microsoft Excel was provided as a spreadsheet for graphics and analysis of data. Documents appeared virtually identical to those in the hard copy NDA submission. Searching the text was facilitated by the use of buttons on the screen, which allowed the NDA to be searched for a particular term. Data could be located either in WordPerfect documents, or in an Oracle database (using Andyne GQL) by querying the data. The data queries could be performed ad hoc, in which the reviewer selected all the parameters for a search, or with predefined query buttons, which retrieved data for principal treatment-related changes. This type of system also could serve as a useful model for both in-house nonclinical review and the submission of INDs and IND amendments. PMID- 7494904 TI - Quick estimate of the regulatory virtually safe dose based on the maximum tolerated dose for rodent bioassays. AB - With a limited subset of National Cancer Institute/National Toxicology Program (NCI/NTP) bioassays, Gaylor (Regul. Toxicol. Pharmacol. 9, 101-108, 1989) showed that the regulatory virtually safe dose (VSD), corresponding to an estimated lifetime cancer risk of less than 10(-6), could be estimated within a factor of 10 simply by dividing the maximum tolerated dose (MTD), estimated from the results of a 90-day study, by 380,000. The purpose of this current study was to extend the analysis to all carcinogens in the Carcinogenic Potency Database (CPDB) utilizing the TD50 (average daily dose rate in mg/kg body wt/day that was estimated to halve the probability of remaining tumor-free at a specified tissue site throughout a 2-year study). Using the relationship between the upper bound on the low-dose slope (q1*) and the TD50 reported by Krewski et al. (Risk Anal. 13, 383-398, 1993) and the ratio of the maximum dose tested (Max-D)/TD50 obtained in our present analysis, an estimate of the regulatory VSD was given by the MTD/740,000, for NCI/NTP rodent carcinogens. This was about a factor of two lower than the limited analysis conducted by Gaylor. There was little difference when the chemicals were divided into mutagens and nonmutagens. Ninety-six percent (134 of the 139 NCI/NTP rodent carcinogens) of the regulatory VSDs calculated from the individual TD50s obtained from the 2-year bioassays were within a factor of 10 of the MTD/740,000.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7494903 TI - Validation of a modified 28-day rat study to evidence effects of test compounds on the immune system. PMID- 7494905 TI - Using the EPC approach to develop health-based soil and water screening concentrations for environmental chemicals. AB - Reference doses (RfDs) and reference concentrations (RfCs) developed by the United States Environmental Protection Agency (USEPA) are typically used to assess the risk of potential adverse human health effects from exposure to environmental chemicals. For a large number of chemicals, however, USEPA RfDs or RfCs have not yet been determined. Thus, for risk assessments that involve a large number of chemicals, there may not be USEPA- or state-derived toxicity constants for all chemicals present at a particular site. A potential solution to this problem is to use estimated permissible concentrations (EPCs) to develop acceptable daily dosages from occupational exposure limits. The EPC is defined as the concentration of a chemical which, under continuous exposure conditions, is expected to be devoid of all acute and chronic toxicities. EPC values are based on allowable occupational exposure limits such as threshold limit values (TLVs), which are workplace exposure guidelines suggested by the American Conference of Governmental Industrial Hygienists. In the present analysis, the EPC method was used to develop interim toxicity constants for more than 30 chemicals which possess TLVs and which appear in the Appendix VIII Hazardous Constituents list, but for which the USEPA has not yet assigned a current RfD or RfC. Appendix VIII chemicals are commonly found at hazardous waste and other sites. Consequently, the development of toxicity constants for the more than 30 chemicals on this list would prove to be most useful for risk assessment purposes. These toxicity constants were then used, together with standard exposure assumptions, to develop acceptable human health-based soil and water concentrations for these chemicals. The use of these values as interim guidance would therefore allow individuals responsible for assessing human health effects posed by environmental contamination to address all site chemicals without performing extensive, chemical-specific toxicological analyses. PMID- 7494906 TI - Chemical mixtures: current risk assessment methodologies and future directions. AB - Some of the most challenging problems that toxicologists confront are determining how biological effects of components in a complex mixture may interact, determining how these interactions affect the overall toxicity of the mixture, and determining how to incorporate this information into risk assessments of chemical mixtures. There has been considerable effort in this area since the publication of the U.S. Environmental Protection Agency's guidelines for risk assessment of chemical mixtures in 1986. This paper reviews the terminology used to describe chemical interactions and the methodologies that have been developed for conducting risk assessments of chemical mixtures. Particular attention is directed towards an examination of the applicability and validity of the methods for the assessment of risk posed by exposure to environmentally relevant concentrations of chemical mixtures. Limited, yet compelling, data are reviewed that suggest that for noncancer endpoints, adverse effects are unlikely to occur when the individual components in the mixture are present at levels well below their respective thresholds. Synergistic or antagonistic effects, not readily predicted from the mechanisms of action of the individual components, are possible when the mixture components are present at levels equal to or above their individual thresholds. Finally, synergistic carcinogenic effects have been observed in animal studies of mixtures, even at relatively low doses. PMID- 7494907 TI - Need to revise the national drinking water regulation for copper. AB - Copper is an essential micronutrient and a potential toxic metal. The USEPA current maximum contaminant level goal (MCLG, a nonenforceable aspirational health goal) and the required treatment technique action level, both set at 1.3 mg/liter of copper are not adequately protective for infants and children under 10 years of age. Infants and children up to 10 years of age have increased susceptibility to copper toxicity. The two main reasons for the high sensitivity in this population subgroup are the presence of a normally high concentration of copper in the liver during early life and the lack of a fully developed homeostatic mechanism of copper in children under 10 years of age. This sensitive group of infants and children represents approximately 17% of the total U.S. population. The adverse health effects caused by drinking water contaminated with copper are abdominal pain, nausea, vomiting, diarrhea, headache, and dizziness. The proposed drinking water guideline of 0.3 mg/liter for copper was derived by the application of the risk assessment procedures approved by the USEPA. Drinking water containing 0.3 mg/liter of copper will provide approximately 26% of the nutritional requirement of copper. This proposed drinking water guideline (0.3 mg/liter) for copper will adequately protect health of infants, children, and adults. PMID- 7494908 TI - Environmental hazards of aluminum to plants, invertebrates, fish, and wildlife. AB - Aluminum is extremely common throughout the world and is innocuous under circumneutral or alkaline conditions. However, in acidic environments, it can be a major limiting factor to many plants and aquatic organisms. The greatest concern for toxicity in North America occurs in areas that are affected by wet and dry acid deposition, such as eastern Canada and the northeastern U.S. Acid mine drainage, logging, and water treatment plant effluents containing alum can be other major sources of Al. In solution, the metal can combine with several different agents to affect toxicity. In general, Al hydroxides and monomeric Al are the most toxic forms. Dissolved organic carbons, F, PO(3)3- and SO(4)2- ameliorate toxicity by reducing bioavailability. Elevated metal levels in water and soil can cause serious problems for some plants. Algae tend to be both acid- and Al tolerant and, although some species may disappear with reduced pH, overall algae productivity and biomass are seldom affected if pH is above 3.0. Aluminum and acid toxicity tend to be additive to some algae when pH is less than 4.5. Because the metal binds with inorganic P, it may reduce P availability and reduce productivity. Forest die-backs in North America involving red spruce, Fraser fir, balsam fir, loblolly pine, slash pine, and sugar maples have been ascribed to Al toxicity, and extensive areas of European forests have died because of the combination of high soil Al and low pH. Extensive research on crops has produced Al-resistant cultivars and considerable knowledge about mechanisms of and defenses against toxicity. Very low Al levels may benefit some plants, although the metal is not recognized as an essential nutrient. Hyperaccumulator species of plants may concentrate Al to levels that are toxic to herbivores. Toxicity in aquatic invertebrates is also acid dependent. Taxa such as Ephemeroptera, Plecoptera, and Cladocera are sensitive and may perish when Al is less than 1 mg.L-1 whereas dipterans, molluscs, and isopods seem to be tolerant. In Al sensitive species, elevated levels (approximately 500 micrograms.L-1) affect ion regulation and respiratory efficiency. Toxicity tends to be greatest near a species' threshold of pH sensitivity. At lower pHs, Al may have a slight ameliorative effect by interfering with H+ transport across membranes. Aquatic invertebrates can accumulate very high levels of Al, but most of this appears to be through adsorption rather than assimilation. Aluminum concentrations may be as high as 5000 mg.kg-1 in insects and greater than 17,000 mg.kg-1 in other invertebrates.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 7494909 TI - Atrazine retention and transport in soils. AB - No pesticide has been studied as extensively as atrazine. The study of atrazine has contributed to our general understanding of the behavior of pesticides in soils. New knowledge and concepts were evaluated, such as atrazine adsorption kinetics, desorption hysteresis, and preferential flow. Corresponding conceptual models were also proposed to explain the behavior of atrazine in soils. Atrazine adsorption-desorption is the major process affecting atrazine behavior in soils and is mainly affected by organic matter and soil pH. Atrazine in soils is subject to biological and chemical degradations. Hydroxyatrazine, the chemical degradation product, is more strongly adsorbed to soil than atrazine. Deethylatrazine and deisopropylatrazine, the major biological degradation products, are more mobile than atrazine. Hydrolysis of atrazine is soil-surface catalyzed and favored by low soil pH. The overall dissipation rate of atrazine was found to be pseudo first-order. Two distinct and different processes are involved in atrazine movement: slow transport through the soil matrix and rapid movement through macropores. The first process is controlled by adsorption kinetics and degradation reactions and can be well explained by models based on chemical heterogeneity, such as the two-site models and second-order models. The second flow process results from preferential flow through large pores and can be explained by physical nonequilibrium models such as the mobile-immobile and two flow domain models. Because both processes coexist in atrazine transport, coupling of physical and chemical nonequilibrium models is often necessary and has shown promise in atrazine transport modeling. However, more efforts are needed in estimating model parameters and in developing management-oriented models. PMID- 7494911 TI - Nocturnal oxygen desaturation and exercise-induced desaturation in subjects with chronic obstructive pulmonary disease. PMID- 7494910 TI - Clinical applications of serum markers for lung cancer. PMID- 7494912 TI - The performance of Mini Wright peak flow meters after prolonged use. AB - The accuracy of 84 new and 35 old Mini Wright peak flow meters were tested using a servo-controlled pump system. The 95% confidence limits for flow measurement across the range of the new meters was between +/- 151 min-1 at the lower end of the range and +/- 28 l min-1 at the top of the range. The readings for 22 (63%) of the old meters (age range 1-13 yr) were within these 95% confidence limits. For the remaining 13 old meters (age range 1-13 yr) whose readings were not within these limits, there were 11 meters with readings falling below and two meters with readings above these limits. Twelve of these old meters were washed and retested and there was no significant change in their readings. Twenty of the new meters were retested after 1 yr of continuous use and their readings were significantly higher with a median value of 5 l min-1 across the range, although only two of these 20 meters had readings outside the 95% confidence limits set from the 84 new meters. It is concluded that whilst Mini Wright meters aged up to 14 yr can give readings which are as good as new meters, some meters demonstrate significant changes in readings after only 1 yr and washing did not correct this change. It is recommended that clinicians prescribing peak expiratory flow (PEF) meters should be responsible for checking the patient's meter as well as their PEF readings at clinic visits. PMID- 7494913 TI - Oxygen-induced lung injury in the pre-term guinea pig: the role of leukotriene B4. AB - Leukotriene B4 (LTB4) has been reported to promote the formation of lung oedema when infused into the pulmonary circulation of adult animals. The present study evaluated the hypothesis that LTB4 was responsible, in part, for the oedema that develops during oxidative injury of the immature lung. Significant increases were found in LTB4 concentration in bronchoalveolar lavage fluid obtained from pre term guinea pig pups maintained in 95% oxygen for 48 h (P < 0.05) and 72 h (P < 0.05) compared to pups maintained in 21% oxygen. Cellular analysis of lavage fluid revealed a concurrent influx of neutrophils into the hyperoxic-injured lung at these times. The protein concentration of lavage fluid was also increased by 48-h hyperoxia exposure indicating elevated pulmonary microvascular permeability. In a second series of experiments, pups exposed to 95% oxygen (and 21% oxygen controls) were treated with a specific LTB4 antagonist (U-75302), at either 0.5, 1.5 or 3.0 mg 100 g body wt to ascertain if LTB4 played a role in either neutrophil recruitment or oedema formation in the immature lung. The number of neutrophils recovered in bronchoalveolar lavage fluid was significantly reduced, compared to vehicle-treated pups, in pups treated with U-75302, at both 1.5 and 3.0 mg/100 g body wt but not 0.5 mg/100 g body wt. Histopathological analysis of pups treated with 1.5 mg U-75302/100 g body wt revealed fewer neutrophils in the pulmonary interstitium (198 vs. 218 mm-2, P < 0.05). The extent of lung microvascular permeability, elevated by hyperoxic exposure, was modulated by increasing concentrations of U-75302. Specifically, treatment with 0.5, 1.5 and 3.0 mg U-75302/100 g body wt reduced microvascular permeability by 17, 67 and 98%, respectively. In conclusion, LTB4 plays an important role in oedema formation in acute oxidative injury of the immature lung and this is mediated, in part, through neutrophils. PMID- 7494914 TI - Sufficient indication of nocturnal oxygen saturation and breathing pattern in COPD patients, from a single night's study. PMID- 7494915 TI - Use of the Epworth Sleepiness Scale to demonstrate response to treatment with nasal continuous positive airways pressure in patients with obstructive sleep apnoea. AB - The Epworth Sleepiness Scale (ESS) was used to measure the degree of daytime sleepiness in two groups of patients with obstructive sleep apnoea (OSA), before and after treatment with nasal continuous positive airways pressure (CPAP). One group (50 patients) were assessed after 2 months CPAP treatment after which the mean ESS fell from 16.4 [standard error of mean (SEM) 0.52] to 7.0 (SEM 0.56). A second group (25 patients) were assessed after 1 yr of treatment: a similar fall in mean ESS was seen from 15.2 (SEM 1.13) before treatment to 6.0 (SEM 0.72). These results imply that the ESS can be used clinically to demonstrate the response of daytime sleepiness in OSA to treatment with CPAP, and that the fall in ESS seen after 2 months is sustained after 1 yr of treatment. It is also possible that this approach could be used to monitor the progress of treatment with CPAP. PMID- 7494916 TI - Diagnostic role of fiberoptic bronchoscopy in suspected smear negative pulmonary tuberculosis. AB - Fiberoptic bronchoscopy with bronchoalveolar lavage (BAL) and transbronchial biopsy (TBB) was performed in 40 patients suspected to have pulmonary tuberculosis, in whom chest roentgenogram revealed minimal infiltration and sputum smears were negative for acid-fast bacilli. Bronchoscopic procedures provided overall diagnostic yields in 47.5% (19/40) of patients. The diagnostic yield of overall bronchoscopic procedures for tuberculosis in this study was 32.5% (13/40) of patients. It consisted of positive BAL smear in 7.5% (3/40) of patients, positive for mycobacterial culture in 15% (6/40) of patients and TBB revealing granuloma in 17.5% (7/40) of patients. Non-tuberculosis conditions were diagnosed by the bronchoscopic method in six patients (15%). These results suggest that in an area with a high prevalence of tuberculosis, bronchoscopic procedures should be performed in those cases in which other diagnoses such as malignancy must be ruled out. Transbronchial biopsy has a major role for early diagnosis and should be performed in all cases, if possible. PMID- 7494917 TI - Orthodeoxia and platypnoea after acute organophosphorus poisoning reversed by CPAP: a newly described cause and review of the literature. AB - The case of a 60-year-old male patient, who survived severe organophosphorus poisoning, and subsequently developed platypnoea and orthodeoxia is described. The patient was mechanically ventilated for a long period of time in the intensive care unit. During the weaning trial, he developed platypnoea and orthodeoxia (PaO2 85 mmHg in recumbency, and 40 mmHg in upright position). Interestingly, the patient's orthodeoxia was alleviated on continuous positive airway pressure (CPAP) treatment. This is a newly described cause of the platypnoea-orthodeoxia syndrome. The possible pathophysiological mechanisms are discussed and a review of the reported abnormal states associated with this condition is presented. PMID- 7494918 TI - Spontaneous haemothorax as a complication of anti-coagulation following coronary angioplasty. AB - Haemorrhagic complications are well recognized when heparin is used during percutaneous transluminal coronary angioplasty (PTCA). We present a 74-year-old female who developed a large acute spontaneous haemothorax 20 h after coronary angioplasty. Spontaneous haemothorax has rarely been described and is predominantly associated with pulmonary embolism. In the absence of a pulmonary embolus, a spontaneous haemothorax during anti-coagulation has only been described twice in the world literature since 1862 (1). This is the first description of this complication following the use of heparin during PTCA. PMID- 7494919 TI - Cor pulmonale due to untreated asthma in a young woman with coexistent anorexia nervosa. PMID- 7494921 TI - Neurological complications in psittacosis: a case report and literature review. PMID- 7494920 TI - Ulcerative colitis responsive to smoking and to nicotine chewing gum in a patient with alpha 1 anti-trypsin deficiency. AB - Ulcerative colitis is one of the few diseases in which smoking appears to confer some benefit (1). We report a patient whose ulcerative colitis deteriorated on several occasions on stopping cigarettes, and improved on restarting smoking. As a result, she continued smoking despite developing airflow limitation and severe emphysema. She was subsequently found to have alpha 1 anti-trypsin deficiency. She later noticed that she could get a similar benefit in her colitis with nicotine chewing gum as she had with cigarettes. For patients with smoking responsive ulcerative colitis, non-tobacco forms of nicotine delivery such as gum or transdermal patches should be considered to avoid the hazards of cigarette smoke. PMID- 7494922 TI - Anti-Jo-1 syndrome presenting as cryptogenic organizing pneumonia. PMID- 7494923 TI - [1st Symposium on Heart and Hypertension. Madrid, Spain, October 1994. Proceedings]. PMID- 7494924 TI - [The polymetabolic syndrome. The experiences of the DRECE study. Dieta y Riesgo de Enfermedades Cardiovasculares en Espana]. AB - Different risk factors frequent in NIDDM predispose to suffer an early atherosclerosis. Among these and beside lipidic disorders (hypercholesterolemia, hypertriglyceridemia and low HDL-cholesterol levels), hyperinsulinemia and/or insulin resistance, obesity and arterial hypertension are placed. In Spain, 135,000/year deaths from cardiovascular diseases (42% of total deaths) are produced. To plan a preventive strategy requires the knowledge and previous analysis among the population of the situation with regard to risk factors. In order to know in Spain the actual prevalence of cholesterol and other lipidic fractions and their relationship with different dietetic habits and other cardiovascular risk factors, a study on "Diet and Cardiovascular Risk Diseases in Spain": DRECE, was performed. Remains to be explained in the mediterranean countries the low incidence of coronary morbid/mortality, in spite of a high fat consume and cholesterol levels. In this direction, the role of a few micronutrients and other common elements in our diet, some of them with antioxidant properties, may have as protector agents in the atherosclerotic ischemic heart disease process, acquires interest. PMID- 7494925 TI - [Hypertensive cardiopathy. Studies on left ventricular hypertrophy]. AB - This paper reviews the work of the cardiology Service of the Hospital de Basurto de Bilbao on hypertensive cardiomyopathy, diastolic disfunction and exercise tolerance, presenting a new pathophysiologic classification and the possibility of altering the underlying functional factors. The paper also reviews the presentation and evolution of microvascular angina in hypertensive patients. PMID- 7494926 TI - [Arterial hypertension in Spain]. AB - This paper is the written version of the conference that under the same title was presented in the XXX Congreso Nacional de la Sociedad Espanola de Cardiologia. In the paper, the topic of arterial hypertension (AHT) in Spain is approached from different perspectives, as well as to what extend cardiologists influence or are influenced by it. First, there is a brief overview concerning the diagnostic and therapeutic status of AHT in the population, and its evolution in the last years. The paper continues analyzing the research on AHT sponsored by the Fondo de Investigacion Sanitaria in recent years, and emphasizes the little participation of cardiologists as principal investigators in this area. Next, publications in Spanish and international scientific journals are assessed. To end the paper, a proposal of the possible role of the cardiologist on diagnosis, treatment, continued education and research on hypertension is presented. PMID- 7494927 TI - [Complementary tests: conventional and Holter electrocardiography]. AB - Arterial hypertension can cause left ventricular hypertrophy and consequent hypertensive cardiopathy, a condition that is associated with the development of heart failure, arrhythmia and ischemia. Electrocardiography is the oldest and simplest method for diagnosing left ventricular hypertrophy: nevertheless, though established electrocardiographic criteria are highly specific, their sensitivity is low, far below that of echocardiography. There are, however, new electrocardiographic criteria with improved sensitivity; moreover, the presence of electrocardiographic signs of hypertrophy in a hypertensive patient makes a diagnosis of hypertrophy highly probable. The Holter monitor has many applications in the assessment of hypertensive patients. It identifies auricular and ventricular arrhythmias in terms of anatomical patterns, and can be used to evaluate drug response and detect the presence of silent ischemia and sudden death. In conclusion, electrocardiography is an easily accessible tool and a highly useful one in the management of hypertensive patients, in spite of its limitations. PMID- 7494928 TI - [Ergometry in the diagnosis and prognosis of arterial hypertension]. AB - Blood pressure is a dynamic parameter and stress testing constitutes a diagnosis and prognostic method, usually little used, but it lets you know if blood pressure control during exercise is correct. Several studies have demonstrated the predictive value of arterial hypertension when resting, when exaggerated blood hypertension exists during exercise. During the present paper last studies are reviewed about this issue in the world press. We can extract the following conclusions: Stress testing is useful, in general, for the diagnosis, prognosis and therapeutic control of the arterial hypertension besides higher reproducibility and existing a good correlation between maximum blood pressure during exercise and the left ventricular hypertrophy. PMID- 7494929 TI - [Diagnostic methods and the prognosis of the heart-hypertension interaction. Isotopes]. AB - The study with isotopes of the hypertensive cardiopathy permits to value the systolic and diastolic dysfunction, and the myocardial ischemia. It is accomplished by means of the equilibrium ventriculography, employing erythrocytes marked with 99mTc. Would be indicated in those cases in which it would not be possible a good definition of the echocardiographic image. The study of the images obtained with 201T1, at rest and in exercise, permit to evaluate the myocardial ischemia and coronary reserve, in hypertensive patients with normal epicardial coronary arteries. PMID- 7494930 TI - [Diagnostic methods and the prognosis of the heart-hypertension interaction: cardiac echo-Doppler]. AB - Among all the cardiovascular risk factors, Left Ventricular Hypertrophy (LVH) is the most important and its diagnosis by echocardiography is essential in routine evaluation of Arterial Hypertension (AHT). Calculation of the cardiac mass and of the relative thickness of the left ventricular wall are necessary steps in morphological classification of the heart of AHT into four subtypes: normal, concentric remodeling, concentric and eccentric hypertrophy. Study with M and 2D echo can be complemented by the analysis of systolic function and ventricular afterload. Alteration of the diastolic function by pulsed Doppler is sometimes the first sign of cardiac dysfunction in AHT. The three classic patterns of mitral filling, prolonged relaxation, restrictive and pseudonormal, often give only a rough analysis. The diversity of the factors involved in diastolic dysfunction opens the door to new indicators derived from analysis of flow in the pulmonary veins, especially the relative amplitude of the inverted "a" (ar) and "decalage" between this and the A of the anterograde mitral flow. The "decalage" between the inverted A (Ar) of the left ventricular outflow in relation to the mitral A is the latest approximation in the study of ventricular compliance. The poor overall prognosis which is implied by an increase in cardiac mass is modified by the geometric subtypes, the high morbidity and mortality of concentric hypertrophy being apparently noteworthy. The true prognostic significance of the regression of LVH and the accompanying diastolic dysfunction are still subject to debate. PMID- 7494931 TI - [The clinical utility of the automatic ambulatory recording of the arterial pressure in the diagnosis, prognosis and treatment of arterial hypertension]. AB - Ambulatory blood pressure monitoring (ABPM) is, nowadays, one of the most powerful medical tools, in the evaluation of blood pressure (BP) behaviour during a normal daily life, particularly in hypertensive patients, and to assess with a high level of accuracy, the antihypertensive effectiveness of the pharmacological compounds. In this paper, we present the main indications and limitations of the method, and a new circadian classification of arterial hypertension based on an average deviation index (ADI), resulting of the addition of all the systolic and diastolic ambulatory measurements exceeding 140/90 mmHg recorded along the 24 hour extrahospital monitoring, and dividing the result by two. The study was carried out in 939 hypertensive patients diagnosed by means of basal blood pressure measurements (BP > 140/90 mmHg), and in 45 normal volunteers. Hypothetically, this ADI may vary only from 0 (normal curve) to 100% (complete abnormal circadian curve). From ADI 0 to 100, we have established 4 categories of circadian curves as follows: curve 0 or "normal" if the ADI varies from 0 to 5%, curve type I or "borderline" (white coat phenomenon) if ADI moves between 6 and 30%, curve type II or "diurnal" if the ADI varies from 31 to 65%, and finally curve type III or "sustained", if the ADI goes further on than 66%. The results indicate clearly a high percentage of white coat hypertension (30%) where 4% showed an ADI less than 5%. A half approximately of the patients showed diurnal hypertension, and a quarter of them, both diurnal and nocturnal hypertension, so called sustained hypertension. 75% of all the hypertensive patients showed spontaneous normotensive values at nighttime.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7494932 TI - [Diuretics and beta-blockers as the first option in the treatment of arterial hypertension]. AB - Diuretics have increasingly come to be used for the treatment of arterial hypertension over the last few decades. Although their precise active mechanism is unknown, it is supposed that their usefulness as antihypertensive is a product mainly of their effect on the transport of ions across the cell wall, which reduces peripheral vascular resistance. The adverse metabolic effects of diuretics can be minimized by prescribing the lowest doses presently recommended, or by administering potassium salts or a potassium-retain diuretic. The antihypertensive action of beta blockers is similar to that of diuretics and other antihypertensive drugs, and again, their active mechanism is poorly understood. There are several contraindications for prescribing beta blockers, as well as side effects that must be considered before starting treatment. Well controlled studies on large cohorts have demonstrated the usefulness of both diuretics and beta blockers in preventing the cardiovascular complications of hypertension. These studies have also allowed the medical profession to designate indications for using these drugs in various segments of the population defined by factors of age and race. Given the proved therapeutic efficacy of diuretics and beta blockers, as well as economic considerations, these drugs should be considered first-choice options for the arterial hypertension. PMID- 7494933 TI - [Controversies in the treatment of arterial hypertension: other drugs as the first step (calcium antagonists, ACE inhibitors and alpha-blockers)]. AB - This paper is the written version of the conference that under the same title was presented in the XXX Congreso Nacional de la Sociedad Espanola de Cardiologia. In the paper, the use of the most recent antihypertensive drugs is advocated as first line therapy. The defence is based in cardiac and non cardiac considerations. Among the former, there are certain advantages of the new drugs in areas as regression of left ventricular hypertrophy and improvement of coronary flow and coronary reserve. Among the non cardiac considerations there are some metabolic factors (non adverse effect on the lipid profile), as well as a positive effect on some endocrine-metabolic cascades like the insulin resistance syndrome and a very important role in certain special situations as coexistence with other diseases and individual characteristics of the patient (pregnancy or elderly for example). PMID- 7494934 TI - [Arterial hypertension in the elderly. The outstanding aspects]. AB - Arterial hypertension (AHT) is one of the principal health problems facing the community today. Because the prevalence of AHT among the elderly is high and management of the condition is problematic, concern is increasing as the population ages. This article provides the following: a) a short summary of epidemiological data, the most important being the criteria used to define the disease, which has a prevalence of around 50% in patients over 65 years of age (and higher among women); b) a discussion of the role of AHT as a risk factor in the elderly; c) a review of the most important findings and conclusions from studies carried out among the elderly, and d) some final considerations toward the eventual formulation of treatment guidelines. PMID- 7494935 TI - [The pathogenesis of the evolution of the atheroma plaque]. AB - The different theories on the pathogenesis of the atherosclerotic plaque may be integrated into a single multifactorial one. According to this, the atherosclerotic risk factors produce impairement of the endothelial cell function as well as the release of growing factors leading to smooth muscle cells and macrophage activation and subsequent collagen synthesis and foam cells formation, the two main components of the atheroma plaque. The arterial thrombosis is another important factor for the atherosclerotic plaque growth and for the occlusive complications that leads to acute ischemic syndromes. The same etiologic risk factor that initiate the atherosclerotic process are implicated in its progression. Most coronary acute syndromes show the same pathologic syndrome of a fissured atherosclerotic plaque with a superimposed thrombus. The lipid rich plaques with a thin fibrous cap and the presence of monocyte/macrophages are unstable and prone to fissuring more than the old fibrous plaques. The arterial stress can be an important precipitating factor for the plaque disruption. The possibility of slowing human atherosclerosis and even inducing its regression is one of the present therapeutic goals. One of the most useful approaches is the lipid lowering therapy in hyperlipidemic patients with atherosclerosis. These treatments have demonstrated its efficacy in decreasing the arterial occlusive complications even before showing the atherosclerotic plaque regression. Another approach would be the use of aspirin and other antiplatelet and antithrombotic agents that may partially prevent the progression of the atherosclerotic process. PMID- 7494937 TI - [The epidemiology and risk factors of ischemic cardiopathy]. AB - The prevention of cardiovascular diseases has been considered the first cause in diminishing coronary heart disease mortality. The main objective is the control of risk factors in primary as well as in secondary prevention. The correlation between diet rich in saturated fat and plasma cholesterol was the first milestone in this field followed by the improvement in cholesterol levels and subsequently in CHD mortality. Smoking cessation, physical exercise and control of blood pressure have shown to be efficacious. May be that Syndrome X that corresponds to insulin resistance, central obesity and low levels of HDL among other characteristics, reaches in the future a greater importance in CVD epidemiology. Other risk factors like those of psychosocial origin, thrombogenesis or hormones need more research in order to know their actual importance. PMID- 7494936 TI - [The relationship between clinical events and the angiographic lesions in coronary atherosclerosis. Hypolipemic treatment and plaque stabilization]. AB - Secondary prevention studies have shown that lipid-lowering therapy improve angiographic outcome and reduce mortality and incidence of ischemic clinical events in patients with coronary artery disease. The mechanism responsible for the improvement in prognosis seem to be subtle as this improvement cannot be explained by changes in the angiographic diameter of coronary arteries. Atherosclerotic plaque rupture and subsequent thrombosis are the central features in the pathogenesis of acute coronary events: unstable angina, myocardial infarction and sudden death. Cholesterol lowering might decrease the risk of plaque rupture and its thrombogenicity, as well as normalise the impaired endothelial function in hypercholesterolemic patients. In this report, the effects of lipid-lowering therapy on angiographic outcome and incidence of clinical events, in patients suffering from coronary artery disease, are reviewed, and the mechanisms that might explain these findings are discussed. PMID- 7494938 TI - [The pharmacological treatment of hypercholesterolemia]. AB - Hypercholesterolaemia therapy should begin with implementation of a suitable diet in order to achieve optimum weight and reduce the intake of saturated fats. Often, however, a dietary regime is not sufficient to decrease cholesterol levels in hypercholesterolaemic patients and drugs must also be used. Some of the pharmacological options available act principally on LDL cholesterol, whereas others have more effect on triglyceride rich particle. HMGCoaA reductase inhibitors exert the greatest effect on plasmatic LDL cholesterol levels and are therefore recommended in cases of moderate or severe hypercholesterolaemia, particularly in the secondary prevention of ischaemic cardiopathy. Comparing the three statin drugs, lovastatin, pravastatin and simvastatin, the latter has recently been shown in study 4S to be effective in reducing global and coronary mortality in patients with a history of coronary heart disease. In addition to their action on LDL cholesterol, these drugs also increase HDL cholesterol, reduce triglycerides and have a beneficial effect on some of the fundamental mechanisms involved in the development of arteriosclerosis. Ion-exchange resins moderately reduce cholesterol levels, thus are used in young people or in combination with other drugs when a further reduction of LDL cholesterol is required. Their main drawback, however, is that they can often cause digestive intolerance. Primary prevention trials have shown that fibrates also reduce mortality by coronary heart disease but have no effect on global mortality. They are well tolerated and are used in the treatment of mixed hyperlipaemia. Other products, such as probucol and oestrogens, are also used but only under specific circumstances. PMID- 7494939 TI - [The Scandinavian Simvastatin Survival Study: the clinical consequences]. AB - In the recently reported Scandinavian Simvastatin Survival Study (4S), 4,444 men and women between 35 and 70 years of age who had coronary heart disease and plasma total cholesterol concentrations of 212 to 310 mg per deciliter (5.5 to 8.0 mmol/l) received simvastatin or placebo for 5.4 years; in the treatment group, mortality from coronary heart disease was 42 percent lower and mortality from all causes was 30 percent lower. Further analysis of the relation between the risk of major coronary events and baseline cholesterol levels in patients randomised to placebo or simvastatin therapy in the study showed that simvastatin significantly reduced the risk of major coronary events in all quartiles of baseline total, high-density-lipoprotein, and low-density-lipoprotein cholesterol, by a similar amount in each quartile. Clearly, cholesterol-modifying interventions can substantially improve the outlook for patients with coronary heart disease and should be considered in identification and treatment of patients with established coronary heart disease. The available data from secondary intervention trials indicates that all patients with documented ischemic heart disease should be offered a reliable assessment of plasma total cholesterol. However, since the impact of simvastatin on CHD seems to begin after about one year of therapy it would be reasonable to exclude very old persons and patients with serious disease with a limited life-expectancy. As a consequence of the 4S data it should be recommended that all the patients in question, who after lifestyle modifications and non-pharmacological risk factor modification still have total cholesterol levels above 200 mg per deciliter (5.2 mmol) should be offered treatment with a HMG-CoA reductase inhibitor.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7494940 TI - [The effect of a reduction in plasma cholesterol on the evolution of coronary disease]. AB - Elevated serum cholesterol level has a causal role in the genesis of coronary atherosclerosis and causes plaque activation because it leads to plaque rupture, increases thrombus formation and adversely influences the function of endothelial cells. In patients with evidence of coronary heart disease (angina pectoris, previous myocardial infarction or previous coronary revascularization) the overall effect of cholesterol reduction therapy on the progression of lesions is modest. Nevertheless, the results of secondary prevention trials provide evidence that a reduction in the level of cholesterol leads to a significant decrease in the rate of cardiovascular events, in the rate of new procedures of revascularization by means of coronary surgery or angioplasty, in the coronary heart-disease-mortality and in the non-coronary-heart-disease mortality. These effects probably mean some benefit on function, vulnerability and thrombogenicity of the plaque. In patients with previous revascularization procedures interest of secondary prevention by means of cholesterol lowering must be special, because in them the probability of long-term success should be optimized for the maximum patient benefit and the best use of health-care resources. PMID- 7494941 TI - [The practical aspects of lipid management in secondary prevention]. AB - Secondary prevention of ischaemic heart disease includes the important aim of reducing plasma lipids, as have been suggested by several recent trials focusing both on plaque regression-stabilization and on mortality (4S). The degree of acceptance of recommendations of the different panels of experts, however, has been low. Implementation of these secondary prevention measures in clinical practice requires the understanding of their pathophysiologic and epidemiologic basis, as well as an appropriate spread of experts recommendations and a good relationship of the different levels of health care to cardiac patients. PMID- 7494942 TI - The revision of a workload measurement tool to reflect the nursing needs of patients with traumatic brain injury. AB - Rehabilitation for persons with head injuries is a multifaceted process that requires structure, consistency, and predictability. This article discusses the process followed by nurses on one rehabilitation unit to revise a standard nursing workload measurement tool to reflect the physical, cognitive, behavioral, and psychosocial needs of patients with traumatic brain injury (TBI). Family education and support, a critical component of care, was also addressed. The flexibility of the tool, known as GRASP, allowed the incorporation of neurobehavioral items based on the Rancho Los Amigos cognitive functioning scale and the inclusion of important therapeutic nursing interventions. The revised tool reflects the time required to deliver comprehensive nursing care to TBI patients and, thus, promotes the desired rehabilitation outcomes for patients and their families. PMID- 7494943 TI - Quality benchmarks for brain injury rehabilitation services. AB - Quality, a leading priority in health care today, must be quantified if a rehabilitation facility is to accurately evaluate its treatment effectiveness and durability. Quality can be seen as including four domains, or dimensions: accountability, professional staffing, value, and proven results. This article offers suggestions to rehabilitation case managers and discharge planners, who are in search of quality, on how to evaluate a brain injury rehabilitation program prior to referring patients to a facility. PMID- 7494944 TI - Clinical nursing implications for the recovery of atrophied skeletal muscle following bed rest. AB - Atrophied skeletal muscle is a common clinical manifestation of bed rest that occurs primarily because of a lack of weight-bearing activity on the muscle. Findings from several studies using an animal model to simulate the effects of bed rest on muscles indicate that when atrophied muscle is used again for weight bearing activity, it undergoes a series of physiological changes, such as muscle fiber (myofiber) damage, death, and regeneration. Also, in the recovering muscle, the inflammatory white blood cells, called macrophages, accumulate and shift in type in relation to the ongoing myofiber changes. Similar processes may occur in the muscles of a patient resuming normal physical activity following bed rest. The authors briefly describe the physiological changes related to atrophied muscle recovery and the implications for nursing care. Nursing measures for the recovery period may include (a) assessing for symptoms associated with muscle injury, decreased strength, and fatigue; (b) encouraging sufficient protein intake and maintaining normal metabolic demands to ensure muscle repair; and (c) temporarily avoiding the use of immunosuppressive therapy, if possible, to ensure adequate macrophage function. PMID- 7494945 TI - Chronic childhood illness: a nursing health-promotion model for rehabilitation in the community. AB - In this article, the authors present a cultural model for community practice, which can be used by an interdisciplinary healthcare team caring for children in the community. Its focus is on rehabilitation issues related to the patient and the family members who are coping with a child who has a disability related to chronic illness. The need for culturally sensitive care that empowers the patient and the family and that brings the healthcare team and the community together is stressed. The current healthcare situation of many poor Americans in urban areas also is discussed, along with creative strategies for nurse-managed, advanced practice interventions. PMID- 7494947 TI - Developing a patient classification system for the pediatric rehabilitation setting. AB - Patient classification systems can be valuable tools for identifying trends in acuity and for justifying budgeted nursing care hours. A valid and reliable patient classification system can greatly facilitate the delivery of cost effective, high-quality care. However, the more highly specialized the institutional population, the more difficult it becomes to use a standardized system. In this article, the authors address the development of a patient classification system for a postacute rehabilitation pediatric facility. Included is a discussion of the various pilot phases, the implications for clinical nursing practice, and the impact on family-centered care. PMID- 7494946 TI - What happens after a traumatic brain injury?: four case studies. AB - This article reports on four women with traumatic brain injury (TBI). It describes what happened to the women during their initial trauma and what life was like for them following their injuries. TBI can have devastating lifelong consequences because the brain is the body's central functioning unit. Damage from a TBI trends to be comprehensive, extending beyond physical injuries and changing the way a person behaves and feels. It can even bring about a total change in personality and often leads to psychosocial problems. Family and friends of the person with TBI may have difficulty understanding and coping with these changes. The authors suggest nursing interventions to help both the patient and others deal with TBI. PMID- 7494949 TI - You can't judge a book by its cover: lessons I learned from B.C. PMID- 7494950 TI - Rehabilitation nursing in China. PMID- 7494948 TI - From within. PMID- 7494951 TI - [Overview of nosocomial infections in a neonatal intensive care unit (1989 1993)]. AB - Newborns admitted to any Intensive Care Unit (ICU) are more at risk for nosocomial infections. The author analyzed the incidence of nosocomial bacteriaemias occurred in the neonatal ICU of Trento, especially focusing on the relationship between weight at delivery, incidence of infections and mortality. The incidence of bacteriaemiae during the observation period (1.189-31.12.93) accounted for 3.5%, and 2.8% were classified as nosocomial. The most frequently involved organism was the Staphylococcus PCN (62% of bacteriaemiae). High risk newborns weighted < 1500 g; 10.7% were affected by nosocomial bactaeriaemia, representing the 56.1% of all the observed bacteriaemiae. A strong association (p < 0.05) was also observed between bactaeriaemiae and mortality, in newborns weighting less than 1000 g. Since low and very low birth weigh infants are particularly at risk for infections, special attention should be paid to the invasive techniques and a systematic reporting and documentation of epidemiology of infections, in order to monitor impact of modifications in caring techniques is warranted. PMID- 7494953 TI - [Research methods and instruments]. AB - Research papers published on some of the most well known medical and nursing journals are presented and discussed. The main aim of the contribution, which opens a new arena for discussion on the Rivista dell'Infermiere is to critically appraise published research works focusing both on strengths and novelty as well as weaknesses in the hypothesis formulation, methods and instruments used, discussion of results. A critical analysis should enable nurses to start learn to read and eventually write a research protocol, possibly avoiding some common mistakes. PMID- 7494952 TI - [Public opinion and transplantation: results of a survey]. AB - General information on organ transplant is generally lacking and the issue very often arouses feelings of fear and suspicion. Aim of the survey was to assess the level of information of general population on legal, ethical aspects and other topics related to organ donation and transplant. A questionnaire was administered to a convenience sample of 700 inhabitants of Gallarate (MI). The open and closed questions explored areas such as the organ donation from a living person, xenotransplants, the role of media. 520 questionnaires (74.3%) were returned. There is a positive attitude toward organ donation but the specific knowledge of the large majority of people is very rough and superficial: for instance, only 36% of respondents knew the difference between organ donation from a living person and a corpse. The lack of correct information was considered the main reason (66%) for the insufficient number of organ donations. PMID- 7494954 TI - [Evaluation of patient satisfaction: limits and potentials]. PMID- 7494956 TI - [Children bought and sold...]. PMID- 7494955 TI - [Selection of contraceptives]. PMID- 7494957 TI - [International summits and conferences of the United Nations: for whom?]. PMID- 7494958 TI - [Prevention of prematurity]. AB - Premature delivery is defined as birth between 22 and 37 weeks of pregnancy. Perinatal mortality and morbidity and their sequellae are strongly linked to gestational age. Minimum prematurity is around 3% of births. Medical risks of premature birth should be detected outside of pregnancy (uterine) or during pregnancy, at the first outpatient visit, the critical moment for establishing factors of risk. Preventive measures include awakening women to their responsibility, before and during pregnancy, and suppressing toxic agents (tobacco and alcohol). Treatment of medical causes of preterm birth is more effective if accompanied by measures acting on the environment, such as rest adapted to fatigue, improved environmental conditions, decentralisation and local situation of medical follow-up and social assistance, hospitalisation at home, telesurveillance and, in severe cases, total hospital rest (for which alternatives such as family hotels should be sought). Every element of the existing structures should be used complementarily. PMID- 7494959 TI - [Multiple pregnancies]. AB - Multiple pregnancies do raise the extraordinary problem of their frequency: conceptions of this type have taken on epidemic proportions as a result of the growth of infertility treatment. The study details the obstetric management, neonatal outcome, and follow-up data of women who were delivered of twins, triplets and more. The management of multiple pregnancies includes early diagnosis, meticulous follow-up, and early decrease of maternal activity. Data on the prognosis of triplet pregnancies are of particular importance when the option of selective termination is considered. The most common complication is preterm labor. The prematurity rate is 50% in twin, 95% in triplet and 100% in multiple pregnancy of higher order. The perinatal mortality was 20 to 80 per 1,000 in twins and 40 to 250 per 1,000 in triplets. During last years, a number of changes took place: introduction of medically assisted procreation, early diagnosis via ultrasound scans, considerable increase in the number of cesarean sections in the case of multiple pregnancies, improved prognosis for premature and growth retarded newborns. PMID- 7494960 TI - [Prolonged pregnancies]. AB - Pregnancy prolonged beyond 294 days is not a common situation (less than 6% of all pregnancies). The risks of prolonged pregnancy are fetal macrosomia, postmaturity syndrome, acute fetal distress during labor or meconium release. Close fetal monitoring by clinical examination, cardiotocography and ultrasound scan should be done. If oligohydramnios or abnormal cardiotocogram appear, termination of pregnancy by induction of labor or cesarean section is necessary. Conservative management is possible if antenatal monitoring is normal. PMID- 7494961 TI - [Intrauterine growth retardation]. AB - Intrauterine growth retardation is an important cause of perinatal death. The etiology is predominantly renovascular-syndromes, chromosome or genetic anomalies, and idiopathic hypotrophy. The small for age baby is hypoxic and hypercapnic, with acidosis. Fetal circulation is redistributed to protect the brain. The type of retarded growth (well balanced? or not), its severity and its potential development can be evaluated by echographic examination, and means of Doppler study. The only treatment is fetal removal. The date of this removal is a compromise between the risk of fetal death or neurological consequences of prolonged hypoxia on the one hand, and of the risk of severe iatrogenic prematurity, or neonatal death, if removal is made at too young a gestational age. The only preventive treatment that has been evaluated is lowdose aspirin which, in at-risk patients, has reduced the risk by a factor of four to six. PMID- 7494962 TI - [Management of fetal abnormalities]. AB - Appropriate management of fetal malformations needs confirmation by a tertiary center and search for associated abnormalities and further investigations oriented to causal and prognostic factors. Therapeutic decisions are taken by a multidisciplinary term: pregnancy termination, maternal transfer to a tertiary center, premature delivery or cesarean extraction. Some malformations require immediate surgical treatments, others can be followed up and later corrected. Intrauterine treatments are nowadays evaluated, such as chronic derivations, operative fetoscopy or open fetal surgery. In case of fetal death, a post mortem examination must be performed in order to improve genetic counseling. PMID- 7494963 TI - [Arterial hypertension and pregnancy]. AB - Hypertension is observed in 10 to 15% of pregnancies, but only 10% of affected women will suffer preeclampsia. The pathophysiology of preeclampsia is based on an early anomaly of placental implantation, leading to a cascade of events (secretion of vasopressive prostaglandins, anomalies of hemostatis) which can cause disseminated intravascular coagulation. The diagnosis of preeclampsia is based on hypertension and significant proteinuria (above 0.5 g/24 h), and paraclinical maternal and fetal examinations to follow the evolution and detect the appearance of maternal complications (retroplacental haematoma, Hellp syndrome and eclampsia) and (or) fetal complications (delayed growth, in utero fetal death, perinatal death). The aim of hypertensive treatment is to normalise blood pressure and to avoid maternal complications. Preventive treatment with aspirin reduces the frequency of recurrent preeclampsia and delayed growth of the fetus. PMID- 7494964 TI - [Diabetes and pregnancy]. AB - Pregnancy with maternal diabetes is exceptionally contra indicated. Maternal IDDM and NIDDM required strict glycemic control and must be supervised during all the pregnancy. It is important to early evaluate the risk of gestational diabetes and necessary to have diabetologic, obstetrical and neonatologic managements. In these conditions, pregnancy in diabetes has a good prognostic. These women (IDDM, NIDDM, GDM) must be enrolled in a follow-up program. PMID- 7494965 TI - [Postpartum hemorrhage]. AB - Postpartum hemorrhages are observed in 2 to 10% of all deliveries. They are severe in 1% of the pregnancies. However, they remain a major cause of maternal morbidity and mortality. Post-partum hemorrhages are generally separated in 2 categories. Acute hemorrhage occurs in the 24 hours following the delivery and is mainly caused by uterine atony, retained secondines, placenta accreta, birth canal trauma and uterine inversion. The delayed hemorrhage occurs after 1 day to 6 weeks after the delivery and is often related to uterine infection or abnormal involution of the placental bed. The management requires uterine massage and ocytocine or prostaglandins. Surgical or radiological selective ligation or embolization of the internal iliac arteries can be required. The hysterectomy is proposed in the most severe cases. PMID- 7494967 TI - [Diffuse acute peritonitis. Physiopathology, etiology, diagnosis, development, treatment]. PMID- 7494966 TI - [Maternal transfer and neonatal transfer]. AB - Regionalisation of perinatal health care has not been organised in France. This concept has proposed to reduce mortality and morbidity of newborns of less than 33 weeks or less than 1,500 g at birth by a transfer of the mother before birth to a perinatal center with a neonatal intensive care unit. The available information in France shows that only 14% of those infants will be born in a level 3 centres. In Seine-Saint-Denis district available information shows that the adds ratio for per partum and neonatal death for newborns of 28-36 gestation duration is of 5.27 when a level 1 maternity is compared to a level 3. PMID- 7494968 TI - [Septic shock. Etiology, physiopathology, diagnosis, treatment]. PMID- 7494969 TI - [Complications of myocardial infarction. Diagnosis, development, treatment]. PMID- 7494970 TI - [Hemorrhagic shock. Etiology, physiopathology, diagnosis, treatment]. PMID- 7494971 TI - [Cardiogenic shock. Etiology, physiopathology, diagnosis, treatment]. PMID- 7494972 TI - [Acute angle-closure glaucoma. Etiology, physiopathology, diagnosis, development, prognosis, treatment]. PMID- 7494973 TI - [Dementia syndrome. Diagnostic orientation]. PMID- 7494974 TI - [1/1/96: what does medical insurance pay?]. PMID- 7494975 TI - [The dental health of permanently resident schoolchildren in 16 Zurich rural communities in 1992]. AB - Between 1964 and 1992, caries prevalence (DMFT) of permanently resident school children in 16 communities in the Canton of Zurich decreased by 85 to 87%. In the ten- to fourteen-year-olds, the reduction amounted to 25-29% in the period of 1988 to 1992. In 1992, the fourteen-year-olds had only 1.90 DMFT and 2.49 DFS, the average DS being 0.28. The 21% children with the highest caries experience showed only 9 DFS on average. Such a low caries activity does not favour positive cost-benefit results for individual intensive prevention. From 1964 to 1984 caries prevalence in primary teeth of 7-year-old children decreased to the level of 1.81 dmft; the further reduction to 1.55 dmft in 1992 was not significant. Few primary molars were lost prematurely. According to a saliva-test, 59% of the children aged 10 to 12 with high concentrations of mutans streptococci were free of caries; by comparison, 80% of the children with the lowest mutans-test value, were free of caries. In spite of the easy availability and the multiple usages of fluorides (dentifrices with either 250 or 1000-1500 ppm F, domestic salt, gels with 12,500 ppm F and rinsing solutions with 230 ppm F since 1986-88), only 16% of the children showed signs of dental fluorosis. PMID- 7494976 TI - [Root canal treatment in Switzerland. The quantitative aspects--a determination of its status]. AB - No recent data is available on the number of patients treated for root canal therapy in Switzerland. The aim of this study was to determine, by means of questionnaires, how many and which teeth were root canal treated in Switzerland by private practitioners. One questionnaire concerned the frequency of certain endodontic treatments performed during 10 consecutive working days. 30.1% of the posted questionnaires were correctly completed, returned and evaluated. Per dental practitioner, a total of 4.4 root canal treatments were begun during the 10 consecutive working days. A further 2.2 antiseptic dressings per practitioner were changed during the 10 days. Each dentist completed a root canal therapy on 5.3 teeth, 0.7 re-treatments and 0.1 pulpotomies during the 10 consecutive working days. 40% of all root canal treated teeth were molars which is rather high when compared to data available from other countries. Extrapolating the results to the entire Swiss population, 12.1 patients have a root canal therapy or a re-treatment per year. A similar result (12.9 patients) was recorded for the USA in 1979. PMID- 7494977 TI - [The stress transmission of prefabricated root canal posts]. AB - Photoelastic stress analyses were done with nine prefabricated endodontic posts under axial and diagonal (26 degrees) burden with 5, 10, 15, 20, 25 kg. Stress presented itself in form of interference fringes, which became visible under polarized light. The tapered, grooved pins (Mooser and Unimetric) caused fringes along the shaft and apical as well. They worked at higher stress like a wedge. Cylindrical pins (Para Post Unity 5, Radix-anchor) showed relative strong fringes predominantly at the top. Cylindrically tapered pins (Tenax, CM, Pivomatic) and a conical pin with flat thread and grooves (Exatec-S) caused a more favourable stress distribution and the fringes at the top were clearly slighter. PMID- 7494978 TI - [Approximal and smooth-surface caries. Their diagnosis and therapeutic principles]. PMID- 7494979 TI - [The recognition and treatment of mouth mucosal diseases (II). Hyperkeratosis, leukoplakia, erythroplakia, lichenoid changes and erosions of the mouth mucosa]. PMID- 7494980 TI - [Periodontology--13 years of GTR. A report on the 70th annual meeting of the German Society for Periodontology of 8 to 9 September 1995 in Bremen]. PMID- 7494981 TI - [Geriatric dentistry--an interdisciplinary challenge. A report on the preliminary course and annual meeting of the Swiss Society for the Dental Care of the Handicapped and Aged (SGZBB) and of the Dental Prosthetics Society of Switzerland (ZPGS) of 7 to 9 September 1995 in Zurich]. PMID- 7494982 TI - [The periodontal health of the population and long-term treatment results. Paro Zermatt--a report on the 25th annual meeting of the Swiss Society for Periodontology (SGP) and the annual meeting of the Swiss Society for Dentomaxillofacial Radiology (SGDMFR) of 21-23 September in Zermatt]. PMID- 7494983 TI - [To whom do these teeth belong? Forensic dentistry--the crime of the murder of an unknown young woman]. PMID- 7494984 TI - [Computer Practice Workshop: it no longer works without it. A report on the Computer Practice Workshop of 7 September 1995 in Zurich]. PMID- 7494985 TI - [The risk patient in dental practice. A report on the SSO congress of 18 to 20 May 1995 in Lugano. Societe Suisse d'Odonto-Stomatologie]. PMID- 7494986 TI - [CAR 95. A report on the 9th International Symposium CAR 95 (Computer Assisted Radiology) of 21 to 24 June 1995 in Berlin]. PMID- 7494987 TI - [3 symposia on minimally invasive surgery. A report on the 12th International Congress for Oral and Maxillofacial Surgery of 29 June to 2 July 1995 in Budapest]. PMID- 7494988 TI - [Paro-Zermatt. A report on the 25th annual congress of the Swiss Society of Periodontics (SSP) at Zermatt from 21 to 23 September 1995]. PMID- 7494989 TI - [Clinical and radiologic evaluation of the integrity of medial and lateral collateral ligaments of the elbow in dogs]. AB - Closed reduction is the best therapy in most cases of luxation of the elbow. Clinical retrospective studies showed the presence of arthrosis in 50% of the cases treated with closed reduction (Meyer-Lindenberg et al., 1991) and persisting medial instability after the reduction in more than 50% of the cases. In the present study, the biomechanic of both medial and lateral collateral ligaments was analyzed after selective severing of those ligaments. Severing of the medial collateral ligament led to an average increase of the pronation of 30 degrees with the presence of crepitus on the lateral aspect of the elbow during passive motion. The medial instability of the elbow could be evidenced clinically as well as radiographically. Severing of the lateral collateral ligament resulted in an average increase of the supination of 15 degrees. No evidence of subluxation could be diagnosed using palpation or radiology. Craniocaudal stressed radiographic views of the elbow in pronation are the most helpful to document damages of the collateral ligaments of the elbow. PMID- 7494991 TI - [What is your diagnosis? Idiopathic immunohemolytic anemia in a dog]. PMID- 7494990 TI - [Palliative treatment of tetralogy of Fallot using a PTFE (polytetrafluoroethylene) vascular graft]. AB - The application of a PTFE (polytetrafluorethylene) vascular graft for palliative surgical treatment in a case of tetralogy of Fallot in a dog is described. The surgical procedure took place after an attempt to perform a Blalock anastomosis failed. The surgical technique, the postoperative care and the outcome are presented. The technique and its indications are discussed. PMID- 7494992 TI - [Procedures for health monitoring in a breeding facility for specific pathogen free mice and rats]. AB - Causes of variation in animal experiments include differences in the genotype of the animals as well as a number of environmental factors. Through standardisation of the physical, chemical and biological components of the environment the quality of the results of the experiments can be improved, which in turn leads to a reduction of the number of animals used. One of the means to achieve this goal is the use of specified pathogen free (SPF) animals. To assure the microbiological quality of these animals the population and its environment needs to be screened thoroughly on a routine basis. This publication describes the necessary quality assurance procedures. These include bacteriological, parasitological, virological and histological examinations of the animals themselves, as well as environmental screens such as microbiological examinations of feed, control of water quality or the testing of the efficacy of disinfectants. PMID- 7494994 TI - [First description of a nasal tumor in Nasobema lyricum]. PMID- 7494993 TI - [What is your diagnosis? Key-Gaskell syndrome or feline dysautonomia]. PMID- 7494995 TI - [Developmental trends in farm animal husbandry from an ethological point of view]. AB - The current discussion on "semi-natural" or "extensive" animal production and husbandry shows that such terms are vague and are often used for various meanings. This paper addresses what can be referred to as natural environmental conditions from an ethological point of view, as well as the animal's environment in today's animal production, and how "near to nature" animal husbandry can realistically be. Apart from some outdoor systems with few ethological problems, the predominant part of milk and meat production in the future will still largely take place indoors. Thus, the development of housing conditions which are more appropriate to the animal's needs will still be an important issue of farm animal behaviour science. Consequently the essential environmental requirements of cattle and pigs derived from their species-specific behaviour are described and explained. In the future, an improved fulfillment of ethological requirements will lead to increased group housing in cold and open, structured barns with indoor straw-bedded lying compartments and integrated outdoor feeding and activity areas. This process will also be influenced by ecological, economic and technical aspects. Finally, several studies on cattle and pigs are presented which demonstrate the possible benefits from more appropriate housing systems or the fulfillment of single ethological requirements regarding behaviour, health, fertility and production of the animals. PMID- 7494996 TI - [The use of an externally derived fetal electrocardiogram in obstetrical diagnosis in cattle for the evaluation of fetal vitality]. AB - In 33 pregnant cows a fetal electrocardiogram (ECG) using an external lead was recorded during second stage of labor. A curve of the fetal heart rate pattern was drawn which served to determine the following parameters: basal heart rate, and amplitude as well as frequency of heart rate oscillation. These parameters were compared with APGAR scoring, colostrum intake, blood gas analysis as to their value for evaluation of vitality. There was a statistically positive correlation between basal heart rate during birth and base deficit after birth. Basal heart rate was negatively correlated with plasma pH measured after fetal expulsion. Amplitude of heart rate oscillation was negatively correlated with pH in plasma and concentration of standard bicarbonate, and was positively correlated with carbon dioxide pressure and base deficit. Calves with acidosis at birth or decreased colostrum intake had statistically significantly higher amplitudes of oscillation than healthy control calves. Basal heart rate of the fetus when used as the only parameter might not be suitable in order to exactly evaluate fetal vitality. In contrast, amplitude of heart rate oscillation more reliably indicated a disturbance of fetal condition. Under practical conditions, the described procedure of measuring fetal ECG is suitable in order to detect a viable fetus. PMID- 7494997 TI - Diagnosis of liver flukes in cows--a comparison of the findings in the liver, in the feces, and in the bile. AB - Percutaneous ultrasound-guided cholecystocentesis and aspiration of bile were attempted in 176 cows. These same procedures were performed in another 100 cows immediately after slaughter. The bile samples were examined microscopically for large and small liver fluke eggs. In addition, a fecal sample from each cow was examined for liver fluke eggs. The findings of both groups were summarized, and the results of the fecal and bile sample examinations were compared. In all cows the liver was examined for flukes, and the results were used as a reference. Of 41 cows in which adult flukes were found in the liver, 28 had F. hepatica eggs in fecal samples and 40 had F. hepatica eggs in bile samples. Of 204 cows in which no adult flukes were found in the liver, 23 had F. hepatica eggs in fecal samples and 27 had F. hepatica eggs in bile samples. The sensitivity of the determination of F. hepatica eggs in fecal and bile samples was 68 and 98%, respectively. The negative predictive values for fecal and bile examination were 93 and 99%, respectively. Of 49 cows in which adult flukes were observed in the liver, 13 had D. dendriticum eggs in fecal samples and 44 had D. dendriticum eggs in bile samples. Of 176 cows in which no adult flukes were found in the liver, 19 had D. dendriticum eggs in fecal samples and 49 had D. dendriticum eggs in bile samples. The sensitivity of the determination of D. dendriticum eggs in fecal and bile samples was 27 and 90%, respectively. The negative predictive values for fecal and bile examination were 81 and 96%, respectively. The results of this study indicate that the examination of bile is clearly a more reliable method of diagnosing liver fluke infections than microscopic examination of feces. PMID- 7494998 TI - [Studies of facilities with increased abortion rates in cattle]. AB - Cases of abortions in cattle were investigated using several different laboratory procedures. The purpose was to collect information on an individual animal and on a herd basis that would allow a correct etiological diagnosis and also the institution of prophylactic measures. The cause of the abortion was diagnosed in 30% of the cases and in 16.3% of the herds. Due to the complexity of the problem, there is no routine diagnostic procedure that can universally be recommended and applied. There was no association between the number of tests performed or the number of herd mates included in the diagnostic work-up and the diagnostic success rate. In order to make a more efficient use of the diagnostic procedures, a better understanding of the mechanisms leading to abortion, on an individual animal basis and on a herd basis, is required. PMID- 7494999 TI - How close to closing of academic complexes? PMID- 7495000 TI - Ozone hole--how great the threat? PMID- 7495001 TI - Paraffin poisoning. Partnership the key to prevention. PMID- 7495002 TI - The need for a transformation of the image of the general practitioner in the new South Africa. PMID- 7495003 TI - Contemporary Cuban health care--delivered and undelivered. PMID- 7495004 TI - Paraffin (kerosene) poisoning in South Africa--some errors in a poster designed for black viewers. PMID- 7495005 TI - Africa and the world. PMID- 7495006 TI - Paraffin ingestion--the problem. AB - Paraffin ingestion is the commonest cause of accidental childhood poisoning in South Africa. Children from the lower socio-economic group are affected most. They drink paraffin in the summer months from bottles or intermediate containers, mistaking it for water or cold-drink. The children are predominantly male with a mean age of 24 months. The clinical picture is one of respiratory distress with a hospital case fatality rate of 0.74%. The use of paraffin as a source of household energy in South Africa is on the increase. Based on a modernisation index it would seem that this trend will continue into the next century. It can therefore be expected that the number of cases of paraffin ingestion will steadily increase if no active steps are taken to address the problem. Prevention should entail a wide spectrum of measures, the basis of which should be a child resistant container. An effective durable, low-cost child-resistant container which is easy to pour from should be made available by petroleum companies and/or entrepreneurs and distributed through their network. This should be combined with health education on the danger of paraffin. Health care workers and administrators should be made more aware of the problem and become involved in health education and prevention. Further research should be undertaken on the effect a change in the colour of paraffin and the use of child-resistant caps would have on the incidence of paraffin ingestion in South Africa. PMID- 7495007 TI - The impact of child-resistant containers on the incidence of paraffin (kerosene) ingestion in children. AB - The commonest cause of accidental poisoning in the South African black paediatric population is paraffin ingestion. In this intervention study a specifically designed child-resistant container (CRC) was introduced to evaluate whether its use would decrease the incidence of paraffin ingestion. CRCs were distributed to 20,000 households in the study area (Gelukspan district). No CRCs were distributed in the control area (Lehurutshe district). Health education about paraffin poisoning prevention was given in both the control and the study areas. The monthly incidence rates of paraffin ingestion were monitored during the 14 month intervention period after the distribution and were compared with the pre intervention incidence rates in the study and control areas. The main finding was that the incidence of paraffin ingestion dropped by 47% in the study area during the intervention period. The circumstances surrounding the cases of paraffin ingestion that still occurred in the study and control areas were investigated by means of a questionnaire. We recommend that paraffin be sold in CRCs, and suggestions are made for improving health education to prevent paraffin poisoning. PMID- 7495008 TI - Paraffin (kerosene) poisoning in childhood--is prevention affordable in South Africa? AB - Paraffin (known in some Western countries as kerosene) ingestion is the most common form of acute childhood poisoning in most developing countries. South African black communities reflect a similar pattern, yet the true size and cost of the problem are not known. Without such baseline data the effect of interventions cannot be evaluated. The aim of the study was to determine the incidence and treatment cost of paraffin poisoning in the Cape Peninsula, to identify high-risk areas and to discuss which measures offered the most economical and best preventive potential for this paediatric hazard. A 12-month retrospective study was undertaken in 1990. Relevant patient data were extracted from the files of 6 major Cape Peninsula hospitals. Treatment costs were calculated based on differential hospital costs per inpatient per day, with outpatient costs at one-third of the costs per day. Age-specific rates for affected residential areas were calculated to identify high-risk areas. A total of 436 children (62,5% male), mostly between the ages of 12 and 36 months, were treated at an estimated cost of R111 673. This amount would have been sufficient to provide 95% of households in the 8 identified high-risk areas with child resistant paraffin containers. In these areas age-specific paraffin poisoning rates ranged from 1,8/1,000 to 7,7/1,000. Strategically planned interventions can be cost-effective when weighed against the treatment cost of cases of paraffin poisoning. PMID- 7495009 TI - Trends in photochemical smog in the Cape Peninsula and the implications for health. AB - There has been growing public concern over reports of increasing air pollution in the Cape Peninsula. Attention has been focused on the 'brown haze' and on photochemical smog. Because of deficiencies in the monitoring equipment, information on trends in photochemical smog levels over the past decade is limited. Trends in oxides of nitrogen, one of the main precursors of photochemical smog, and therefore an indicator of the potential for its formation, were examined for the period 1984-1993. Meaningful data for determining trends were available from only a single site. Increases in mean monthly levels, peak hourly levels and the number of times guidelines were exceeded were demonstrated. Given the dynamics of formation of photochemical smog and the particular role of motor vehicles, it is argued that the trends measured at this site are probably an underestimate of the trends in other parts of the Cape Town metropolitan area. Some of the precursors of photochemical smog, notably nitrogen dioxide, and some of its components, notably ozone, have been shown to be detrimental to respiratory health at levels close to, or below, current recommended guidelines. A continuing increase in these pollutants will therefore result in more respiratory illness, particularly among susceptible groups. This calls for an upgrading of monitoring of air pollution in Cape Town and for appropriate steps to prevent its further increase. PMID- 7495010 TI - Comparison of the composition and opsonic activities of imported and South African-manufactured intravenous and intramuscular immunoglobulin preparations. AB - We compared the composition and opsonic activities for two common microbial pathogens (Staphylococcus aureus and Streptococcus pyogenes) of various imported intravenous (IV) (Sandoglobulin, Octagam and Gammagard) and intramuscular (IM) (Beriglobin and Globuman Berna) immunoglobulin (Ig) preparations with those of the corresponding locally manufactured products, Polygam (IV) and Intragam (IM). When tested at equivalent concentrations (1 g/100 ml) the total IgG and IgG subclass concentrations of the various IV and IM preparations were similar. All the test preparations (IV and IM) possessed similar opsonic activity for S. aureus and S. pyogenes. These findings demonstrate that, in respect of IgG content and protective biological activity, Intragam and Polygam, the locally manufactured IM and IV Ig preparations, respectively, compared extremely favourably with the corresponding imported products. PMID- 7495011 TI - Community-based survey versus sentinel site sampling in determining the nutritional status of rural children. Implications for nutritional surveillance and the development of nutritional programmes. AB - A study of the anthropometric status of under-5-year-olds was conducted in the Nqutu district of KwaZulu by means of a representative community-based sample and concurrent samples from primary health care clinics, preschools and primary schools. The first objective of this exercise was to determine the extent of acute nutritional stress in the district as an indication of the appropriateness of food relief efforts in the region. The absence of wasting and the high prevalence of stunting (37.5%) in the community-based sample suggested that the main problem is chronic socio-economic underdevelopment, rather than a severe or immediate lack of food. The fact that fewer than 20% of households are in any way reliant on domestic production for their maize requirements explains why the recent drought has not had a greater impact on the nutritional status of this vulnerable group. This study confirms that the more recent emphasis of the National Nutrition and Social Development Programme on social development is appropriate. An equally important objective of this study was to evaluate the usefulness of clinics, preschools and schools as sites for the collection of anthropometric data and the development of nutritional programmes. This preliminary attempt to develop the methodology for district-based nutrition surveillance suggested that all these sites have limitations both in respect of data collection and community access. The implications of using these sites and the developments needed to improve their usefulness in a future nutrition surveillance system are discussed. PMID- 7495012 TI - Cyclosporin in steroid-resistant nephrotic syndrome. AB - Cyclosporin A (CyA) has recently been shown to be effective in frequently relapsing, steroid-resistant or steroid-dependent childhood nephrotic syndrome (NS). Wider acceptance has been hindered by considerations of cost, toxicity and drug-dependent maintenance of remission. Black children in Africa with NS are mainly steroid-unresponsive and alternative therapies therefore need to be assessed. We report on 8 of 11 children who completed at least 24 weeks of CyA therapy; 6 (2 with membranous nephropathy (MEM) and 4 with focal glomerulosclerosis) showed no improvement, while 2, both with MEM, achieved remission while on CyA therapy and remained in remission for 1 and 3.5 years, respectively, after cessation of therapy. This may have been the natural course of the disease. Of the children who were unresponsive to CyA, 3 died in renal failure 8-30 months after cessation of CyA therapy, 1 had a rising creatinine value when last seen, and 2 were lost to follow-up. CyA trough levels varied between 180 and 875 ng/ml and peak levels between 563 and 1,950 ng/ml. Of 5 repeat renal biopsies, 3 were performed at the end of 24 weeks of treatment and revealed no evidence of CyA toxicity. Two biopsies revealed chronic CyA toxicity. CyA should therefore be used with caution at lower dosages and monitored at currently accepted lower trough levels in children with NS in Africa. PMID- 7495013 TI - The risk of schistosomiasis in Zimbabwean triathletes. AB - A study was carried out to determine the risk of schistosomiasis in triathletes in Zimbabwe. The prevalence of schistosomiasis in 30 triathletes (24 males, 6 females) was compared with that in 24 non-triathlete controls after the 1989/90 triathlon season. All the subjects found to be infected were then treated with praziquantel (40 mg/kg). The seasonal incidence of schistosomiasis in triathletes was then determined in a prospective study during the 1990/91 season. Schistosomiasis was diagnosed by urine and stool microscopy for ova, blood eosinophil counts and serological bilharzial fluorescent antibody tests for IgM and IgG antibodies. There was a significantly (P < 0.05) higher prevalence of schistosomiasis among the triathletes (80%) than among the controls (38%). The seasonal incidence of schistosomiasis was 64%. Exposure of triathletes to fresh water dam swimming in Zimbabwe poses a significant risk for the development of schistosomiasis. PMID- 7495014 TI - Integrins, selectins and CAMs--the 'glue of life'. PMID- 7495015 TI - The herdboy who became head man. PMID- 7495016 TI - [Getting involved in the RDP]. PMID- 7495017 TI - Getting involved in the RDP. PMID- 7495018 TI - [Getting involved in the RDP]. PMID- 7495019 TI - [Critical care--10 years later]. PMID- 7495020 TI - A potentially dangerous design fault in an oxygen analyser. PMID- 7495021 TI - Lethal exposure. PMID- 7495022 TI - Abortion--some practical and ethical considerations. PMID- 7495023 TI - Abortion--some practical and ethical considerations. PMID- 7495024 TI - Abortion--some practical and ethical considerations. PMID- 7495025 TI - Limits of epidemiology. PMID- 7495026 TI - Recent advances in the management of acute leukaemia in adults. PMID- 7495027 TI - Interns' service contracts. PMID- 7495028 TI - Interns' service contracts. PMID- 7495029 TI - Headache and backache after epidural block for postpartum sterilisation. PMID- 7495030 TI - Morphine for cancer pain. PMID- 7495031 TI - [Ischemic heart disease]. PMID- 7495032 TI - [The relation of a number of hormonal indices to risk factors for IHD and its prevalence based on the data from a large population study]. AB - Relevance of IHD risk factors and hormone concentrations (insulin, estradiol, testosterone, prolactin, LH, FSH, TTH and hydrocortisone) to IHD incidence was studied in 1200 men. Multivariate statistical analysis demonstrated that only a small number of hormonal indices are associated with main lipid and nonlipid risk factors. The relationships exist between systolic arterial pressure (SAP) and hydrocortisone, between TTH and total cholesterol, SAP and age, estradiol, testosterone and age. Significant independent associations were found between IHD prevalence and age (R = 0.140, 3 = 0.000), SAP (R = 0.130, P.0.001), HDLP cholesterol (R = -0.082, P = 0.007), height (R = -0.078, P = 0.010), insulin (R = 0.067, P = 0.035), standard deviation of estradiol from median value (R = 0.083, P = 0.017). The findings confirm a great role of high SAP in the development of IHD, a significant and independent association of high insulin levels with IHD prevalence, the presence of nonlinear relation of estradiol concentration to IHD occurrence in males. PMID- 7495033 TI - [The work capacity expertise of motor vehicle drivers suffering from IHD with rhythm disorders and the possibilities for their social and vocational rehabilitation]. AB - The paper presents the results of clinical and labour examinations of 51 drivers with cardiac arrhythmia. Long-term postinfarctional ECG monitoring was attempted in 30 drivers in the course of driving. The frequency of arrhythmia episodes correlated with the presence of extrasystole in acute and subacute period of the disease, anginal functional class, the size and time of myocardial infarction. Single episodes of ventricular extrasystole registered in the course of driving did not interfere with professional activities because such extrasystoles did not imply the risk of more severe arrhythmia. Labour prognosis in drivers with frequent, polytopic ventricular extrasystoles or fibrillation was much worse. PMID- 7495035 TI - [The electrocardiographic diagnosis and clinical course of acute myocardial infarct in patients with an implanted pacemaker]. PMID- 7495034 TI - [The use of tactivin for treating patients with IHD]. AB - Immunomodulating drug tactivin was used for treatment of 29 IHD patients with low T-suppressor activity and high cell sensitization to apoprotein B-containing lipoproteins and with lipoprotein-antibody immune complex detected in the blood. Tactivin was injected subcutaneously in the dose 10 micrograms (5 injections for 12 days). Tactivin treatment normalized T-suppressor activity, lowered concentration of plasma lipoprotein-antibody immune complex and improved clinical course of IHD. The beneficial effect of tactivin was observed in 70% of patients during 2 months. 30% of patients needed a second course of treatment of recover normal immunological and clinical indices. The treatment with tactivin of IHD patients must be combined with application of routine antianginal drugs. PMID- 7495036 TI - [The potentials for the conservative therapy of IHD. Achievements and disappointments]. PMID- 7495037 TI - [The biorhythmological types of work capacity and the late prognosis for patients who have had a myocardial infarct]. AB - The authors analyze postmyocardial infarction 10-year survival and frequency of repeated cardiac accidents (fatal and nonfatal MI and sudden cardiac death). Biorhythmological types of mental and muscular performance were involved in long term MI prognosis. Of 412 males who had their first myocardial infarction at the age under 59, 10-year prognosis was poor for those with early pattern of efficient work vs good prognosis in those who work most efficiently in the evening. The differences in the long-term prognosis of acute myocardial infarction related to performance biorhythm were observed both for subjects performing muscular and mental work. PMID- 7495038 TI - [Magnetic resonance tomography of the heart and major vessels in patients with pulmonary hypertension]. AB - Potentialities of magnetic resonance imaging (MRI) have been studied for evaluation of patients with pulmonary hypertension. For this purpose 54 patients with primary and secondary pulmonary hypertension have been studied using resistive and superconductive MRI systems. MRI proved useful for determination of the cause of pulmonary hypertension (primary, secondary), assessment of the right heart chambers. The cine-MRI enabled detection of tricuspid and pulmonary regurgitation, paradoxical movement of the interventricular septum, thrombi in the pulmonary artery. There were some correlations between MRI parameters and indexes of pulmonary hemodynamics. As a non-invasive imaging modality MRI has a great diagnostic value in characterization of the heart and great vessels in patients with pulmonary hypertension. PMID- 7495039 TI - [The emotional status and indices of cortical neurodynamics in hypertension patients]. AB - At early stages of essential hypertension (EH) clinical psychological examinations of 96 EH patients revealed predominance of anxiety-hypochondriac responses controlled by will. Changes in cortical neurodynamics were evident from a decline in attention concentration. At EH stage II persistent hypertension and prolonged psychogenias gave rise to psychic disadaptation occurring as anxious depressive conditions. Damage to cortical neurodynamics brought about mnestic and thinking disorders. PMID- 7495040 TI - [The circadian profile of the arterial pressure and the structural-functional changes in the cardiovascular system in the initial stage of hypertension]. AB - 24-h blood pressure (BP) monitoring conducted in 52 patients with mild to moderate essential hypertension have shown that the average 24-h systolic and diastolic BP correlates with left ventricular (LV) mass index, while at casual clinical measurements only systolic BP correlates with LV mass index. In non dippers (patients with a nocturnal BP decrease less than 10%) LV hypertrophy is observed 2 times more often than in dippers (patients with a nocturnal BP decrease more than 10%). In the group of dippers 24-h variability of systolic BP and daily systolic BP correlate with LV mass index. Increased variability of night BP correlates with increased venous tone in the calf, increased structural component of vascular resistance and a reduction of blood flow reserve in lower extremities. PMID- 7495041 TI - [The effect of calcium-regulating hormones on the renal transport of calcium in hypertension]. AB - Enhanced and diminished calcemia following administration of parathormone and calcitonin in hypertensive patients, respectively, were found not to differ from relevant values in healthy subjects. Calciuretic effect of parathormone results from its action on the bone and renal calcium transport. Quantitatively, the effects on the bone and renal calcium transport. Quantitatively, the effects on the bone presenting as elevated calcemia and calcium filtration capacity, its excreted fraction and renal excretion prevail, being less pronounced in hypertensive patients. A direct renal effect of parathyroid hormone indicated by stimulation of calcium tubular reabsorption is weaker in hypertensive subjects. Calcitonin administration inhibits tubular calcium reabsorption in the less degree in hypertensive subjects. The disorders in calciuretic function of the kidney in hypertension are secondary to reduced kidney sensitivity to the action of calcium regulating hormones. PMID- 7495042 TI - [The effect of long-term monotherapy with preparations from the 4 basic groups of antihypertensive agents on the quality of life in patients with mild and moderate arterial hypertension. The Multicenter Captopril and the Quality of Life Study. The working group of the Multicenter Captopril and the Quality of Life Study]. AB - A multicenter, controlled, open, randomized trial of antihypertensive and life quality (LQ) effects of monotherapy with captopril vs propranolol, hydrochlorothiazide and nifedipine included 345 males aged 35-60 years with mild and moderate hypertension. The results of 8-month treatment provided the conclusions: the drugs were much alike by antihypertensive efficacy, LQ parameters proved more beneficial in the group on captopril than those on nifedipine and propranolol, hydrochlorothiazide failed to improve life quality, captopril is an effective and safe antihypertensive drug superior to the drugs of 3 other antihypertensive drug groups by positively affecting LQ. PMID- 7495043 TI - [The effect of cilazapril on left ventricular function and myocardial perfusion in patients with essential hypertension (hypertensive disease) stage II]. AB - 22 patients (mean age 45.08 +/- 1.38 years) with essential hypertension stage II were entered in a placebo-controlled blind trial of cilazapril, angiotensin converting enzyme. A complex of clinical-instrumental tests comprised dipidamol test, bicycle ergometry, 199Tl myocardial scintigraphy, echo-CG. A 3-month course of cilazapril reduced myocardial hypoperfusion in 15 patients, improved left ventricular relaxation: Anti-ischemia effect of the drug was also evident. PMID- 7495045 TI - [The antihypertensive activity of benazepril in the long-term treatment of hypertension patients and its effect on adrenal cortical function]. AB - Changes in arterial hypertension, heart rate and adrenocortical hormones (11-OCS, aldosteron, progestins) in the blood and 24-h urine were followed up in the course of 24-week use of angiotensin-converting enzyme inhibitor benazepril (10 20 mg once a day) in 24 patients with mild and moderate essential hypertension (EH) included in a placebo-controlled randomized study. A 2 and 24-week antihypertensive response was achieved in 75 and 71% of patients, respectively. 24-h urinary excretion of corticosteroids before the treatment was increased. After the treatment benazepril reduced excretion of 11-OCS by 42%, but not of aldosteron the levels of which decreased only within the first 2 weeks of treatment. The above trends in changes of gluco- and mineralocorticoid activity should be taken into consideration in long-term treatment of EH with inhibitors of angiotensin-converting enzyme. PMID- 7495044 TI - [The evaluation of cilazapril efficacy in patients with stable arterial hypertension (a randomized study)]. AB - 34 patients with stable arterial hypertension (AH) were randomized by mean arterial pressure (MAP), end-diastolic LV volume and cardiac output in a study of cilazapril efficacy in a dose 2.5 and 5 mg/day. Antihypertensive effect in the majority of patients with MAP 120-135 and 136-150 mm Hg was insufficient (diastolic pressure > 94 mm Hg). Dose-effect action on hemodynamics was absent. Hemodynamic benefits were noted in patients with end diastolic LV volume > 5.0 5.7 cm. PMID- 7495047 TI - [Cardiodynamics and left ventricular myocardial contractile reserve in patients with dilated cardiomyopathy]. AB - Cardiodynamics and contractile myocardial reserves were measured at radionuclide ventriculography and volume loading of the heart (upward position of the legs at 45 degrees) in 43 patients with dilated cardiomyopathy (DCM). The patients displayed a sharp fall in total and regional ejection fractions (BP), stroke volume (SV), cardiac index, a decrease in Vcf and maximum ejection rate (Vmax) in combination with notable cardiomegaly. There were two types of responses to loading test: positive (increased BP, SV, Vcf, Vmax, reduced end-systolic volume) and negative (decreased EP, SV, Vcf, Vmax, increased end-systolic volume) Inotropic effects evidencing the existence or depletion of myocardial contractile reserve. Loading test in DCM patients allows more objective evaluation of the patient's condition and validated therapeutic measures. PMID- 7495046 TI - [Bicycle training by the method of the "free choice" of physical loading in the combined treatment of hypertension patients]. AB - The trends in central hemodynamics and muscular performance were investigated in 107 patients with essential hypertension stage I and II. The intensity of bicycle exercise was either preset (50% of individual exercise tolerance) or decided individually by each examinee (39 and 68 patients of group 1 and 2, respectively). 12 bicycle trainings have improved myocardial contractility at rest and threshold muscular load, exercise tolerance in group 2. Group 1 hemodynamic and performance parameters changed insignificantly. PMID- 7495048 TI - [A comparative analysis of rhythm and conduction disorders in patients with dilated cardiomyopathy and chronic myocarditis]. AB - Rhythm and conduction disturbances were compared for 20 patients with dilated cardiomyopathy (group 1) versus 20 patients with chronic myocarditis (group 2). The diagnosis was confirmed at coronary ventriculography and morphological examination of left and right ventricular endomyocardial biopsies. There were minor differences in the parameters studied I, though supraventricular arrhythmia varied more in group 2. The frequency of ventricular arrhythmia appeared higher in group 2 only in circulatory insufficiency stage 1-IIA. Quantitative differences in ventricular and supraventricular arrhythmias between the groups under moderate circulatory insufficiency may result from different morphological changes in the myocardium. PMID- 7495050 TI - [The dynamic echocardiographic indices after restoration of the sinus rhythm in patients with the compensated type of atrial fibrillation]. AB - The authors observed 19 patients with paroxysmal atrial fibrillation (AF) and compensatory intracardiac hemodynamics accounting for 10% of all their examinees with AF. Following the retaining of sinus rhythm with the help of lanicor, nitrosorbide and lasix patients of group 1 exhibited increased Vs, Vd, Vcf, EF against decreased left atrial size and left atrial index. Patients of groups 2 and 3 in whom the conversion of sinus rhythm occurred due to guanidine or electric impulses, respectively, the above indices changed insignificantly. These patients had depression of left ventricular contractility, increased end-systolic volume, slowing of Vs, Vcf against slight enhancement of left ventricular pump capacity. PMID- 7495051 TI - [Problems in the diagnosis and clinical assessment of atrial dissociation (a report on 2 cases and review of the literature)]. AB - At registration of ECG esophageal leads in two patients with ischemic heart disease an extra atrial rhythm was recorded missed by standard ECG. Similar cases from the literature are considered. PMID- 7495049 TI - [The combined use of the cardiointervalogram and 24-hour ECG monitoring in analyzing heart rhythm disorders]. AB - 2500 records of cardiointervalograms and 24-h ECG monitoring for 1530 arrhythmic patients have been analysed. This complex approach to analysis of cardiac arrhythmia allowed a more accurate differential diagnosis, more detailed visualization of rhythmic abnormalities, more advanced analysis of neurovegetative background of developing rhythmic shifts. PMID- 7495052 TI - [The dynamics of the heart contraction rate and arterial pressure during physical loading after a heart transplant]. AB - Compared to healthy controls, patients with transplanted heart have higher baseline heart rate (HR), delayed initial HR reaction, slower changes of HR both during exercise and after it, diminished chronotropic and inotropic reserves. The data obtained contribute to better understanding of physiology of transplanted denervated heart and objective assessment of patients after heart transplantation. PMID- 7495053 TI - [Prolonged contrast infusion echocardiography with hydrogen peroxide]. AB - A new method of prolonged C-echo-CG with HP implies intravenous administration of 0.3% HP solution for continuous contrasting of right heart chambers for 5-20 minutes. A new technique of C-echo-CG was introduced in 277 patients with innocent cardiac murmur (93), interventricular septal defects (19), interatrial septal defects (26), open oval foramen (42), tricuspid regurgitation (56), pulmonary artery regurgitation (41). Prolonged C-echo-CG with HP was combined with respiratory stress tests. The technique is simple, more effective than short term C-echo-CG, free of severe side effects and complications. PMID- 7495054 TI - [The use of perindopril (prestarium) in arterial hypertension]. PMID- 7495055 TI - [The prognostic importance of echocardiography in myocardial infarct]. PMID- 7495056 TI - Legal issues for nurses. Disciplinary action by the Texas BNE. Part II. AB - This is the second in a two-part series dealing with disciplinary actions by the Board of Nurse Examiners of the State of Texas. Part I discussed the grounds for Board of Nurse Examiners to take disciplinary action against a registered nurse's license. Part II describes the disciplinary process. PMID- 7495057 TI - Advanced practice nursing expands in Texas. Implementing prescriptive authority: 10 steps to guide Texas APNs. PMID- 7495058 TI - Arg506Gln factor V mutation (factor V Leiden) in patients with ischaemic cerebrovascular disease and survivors of myocardial infarction. AB - The point mutation Arg506- > Gln of factor V was recently shown to be an important and relatively common genetic cause of venous thromboembolism. Using a DNA technique based on polymerase chain reaction, we surveyed the blood samples of 236 patients with ischaemic stroke or a transient ischaemic attack, 122 survivors of myocardial infarction and 137 control subjects for the presence of this mutation. Although the frequency of the factor V mutation in patients with arterial disease (4.5%) was not significantly different from that in healthy blood donors (2.9%), a carrier status for this mutant gene was associated with symptoms of migraine and relatively mild angiographic abnormalities among patients with cerebrovascular disease. A more extensive study addressing the occurrence and significance of the mutant factor V mutation in patients with vasospastic cerebrovascular diseases seems to be warranted. PMID- 7495059 TI - Haemostasis factors in angina pectoris; relation to gender, age and acute-phase reaction. Results of the ECAT Angina Pectoris Study Group. AB - The ECAT Angina Pectoris Study is a European multicentre study with the aim of investigating the pathogenetic and predictive role of haemostatic factors in the progression of coronary heart disease. It is the largest study performed up to now with regard to both the number of patients with angina pectoris (n = 3043) and the number of haemostasis assays (n = 23) included. The present paper presents baseline cross-sectional data with particular reference to the relationship of haemostatic factors with each other and with the coronary risk factors age, gender and acute-phase reaction (1). Two clusters of haemostatic factors could be distinguished in which each variable was correlated (P < 0.001) to every other variable: (a) Eight fibrinolysis assays including t-PA, PAI-1 and euglobulin clot lysis time (ECLT), for which PAI-1 appeared to be the dominating factor; (b) antithrombin III, protein C, alpha 2-antiplasmin and plasminogen, the interdependence of which has no obvious explanation. (2). Twelve out of the 23 haemostasis assays were associated (P < or = 0.01) with age. Except for alpha 2 antiplasmin, these relationships indicated an increased tendency to thrombosis with increasing age. (3). Gender differences found in 14 haemostasis parameters do not indicate a consistent difference in the tendency to thrombosis between men and women. Eight haemostasis parameters were on average higher in female than in male patients in the age group over 50 years. (4). C-reactive protein, an acute phase reactant, was positively correlated (P < 0.001) with fibrinogen, factor VIIIc, von Willebrand factor, the fibrinolysis assays t-PA, PAI-1, ECLT and plasminogen.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7495060 TI - Relationships of insulin and intact and split proinsulin to haemostatic function in young men with and without coronary artery disease. AB - Glucose intolerance, hyperinsulinaemia, dyslipoproteinaemia and multiple disturbances of haemostatic function are characteristics of young men with premature coronary artery disease. The relations between glucose, insulin and insulin propeptides, in the fasting state, and after an oral glucose load, and haemostatic function were examined in 62 consecutive non-diabetic men with myocardial infarction before the age of 45 and in 41 age-matched healthy men. "True" hyperinsulinaemia, raised plasma concentrations of insulin propeptides, dyslipoproteinaemia, elevated plasma levels of fibrinogen, factor VII antigen (VIIag) and plasminogen activator inhibitor-1 (PAI-1) activity, and lower antithrombin activity (p < 0.05-0.001) characterized the patients. PAI-1 activity was more closely associated with insulin, intact proinsulin and des 31.32 proinsulin in patients than in controls, whereas the plasma levels of fibrinogen, activated factor VII (VIIa) and VIIag were stronger correlated with insulin and insulin propeptides in subjects without coronary artery disease. Very low density lipoprotein (VLDL) triglyceride was a strong determinant of VIIag and PAI-1 activity levels in both cases and controls, whereas VLDL triglyceride in controls and LDL cholesterol in patients related significantly to the plasma fibrinogen concentration. Thus, our data suggests that plasma insulin and insulin propeptides might be involved in the regulation of plasma fibrinogen concentration, factor VII level and plasma PAI-1 activity. However, apolipoprotein B-containing lipoproteins appear to influence the relationships between insulin, insulin propeptides and haemostatic factors in individuals with premature coronary artery disease. PMID- 7495061 TI - Hypercoagulable state indicates an additional risk factor for atherosclerosis in Werner's syndrome. AB - To assess the risk factors for atherosclerosis in Werner's syndrome (WS), coagulation/fibrinolytic system parameters and lipid levels were investigated in 9 non-smoker patients with WS and compared with normal control values (N). The levels of thrombin antithrombin III complex (p < 0.05), D-dimer (p < 0.05), tissue plasminogen activator (p < 0.005) and PA inhibitor 1 (p < 0.01) were significantly increased, while the level of thrombomodulin (p < 0.005) in the fasting plasma was significantly decreased in the WS cases compared with N. Lipid profiles confirmed that 8 of the 9 patients were of hyperlipidemia type IIb, 7 had hyperinsulinemia and 5 fulfilled the criteria for clinical diabetes mellitus. The hypercoagulable condition suggested the existence of multiple risk factors for atherosclerosis in WS in addition to the previously reported hyperinsulinemia and hyperlipidemia. PMID- 7495062 TI - Raised plasma fibrinogen concentrations in subjects attending a metabolic ward- relation to family history and vascular risk factors. AB - We have evaluated plasma fibrinogen levels in 171 subjects attending a metabolic ward. As in the general population, a significant difference in plasma fibrinogen concentrations (p < 0.05) was found between subjects with diabetes mellitus or hypertension and those without. However, fibrinogen was also abnormally high (p < 0.05) when evaluated according to the presence of a family history of ischemic complications of atherosclerosis (p < 0.05). In this setting, fibrinogen correlated with diabetes mellitus or hypertension as well as with familial risk, and the latter interacted with hypertension (p < 0.05) in accounting for plasma fibrinogen. The relationships between certain fibrinogen genotypes and familial risk have then been evaluated. Analysis of a locus (1.3 kb, HAE III digestion) of the promoter region of the B beta fibrinogen gene, identified a polymorphic cutting site. The allele with the alternative restriction site (H1) was associated with mean fibrinogen levels which were 0.1-0.3 g/l lower than those associated with the other allele (H2). This difference was not statistically significant. No obvious association was found between the familial risk and the presence of the H2 allele. We conclude that in a group of subjects from a metabolic ward, a positive family history for ischemic complications of atherosclerosis is consistently associated with high plasma fibrinogen levels. Interaction with hypertension significantly strengthens the association. PMID- 7495063 TI - In vitro characterization of high purity factor IX concentrates for the treatment of hemophilia B. AB - This study employed sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) analysis and immunoblotting to assess the purity of seven high purity factor IX concentrates: Aimafix (Aima), AlphaNine-SD (Alpha Therapeutic), Factor IX VHP (Biotransfusion), Immunine (Immuno), Mononine (Armour Pharmaceutical), Nanotiv (Kabi Pharmacia), and 9MC (Blood Products Laboratory). The mean specific activity of these products ranged from 68 U factor IX/mg (Aimafix) to 246 U factor IX/mg (Mononine). SDS-PAGE analysis showed that the highest purity product, Mononine, had a single contaminating band under non-reducing conditions. Two additional bands were detected when this product was analyzed under reducing conditions. All other products had multiple contaminating bands that were more apparent under reducing than non-reducing conditions. The immunoblot for factor IX showed a dominant factor IX band for all products. In addition, visible light chain of factor IX was detected for AlphaNine-SD, Factor IX VHP, Immunine, Mononine, Nanotiv, and 9MC, suggesting that the factor IX in these products had undergone partial activation to factor IXa. Another contaminating band was visible at 49,500 for all of the products except 9MC. In addition to this band, high molecular weight contaminants were apparent for some products, most notably AlphaNine-SD. The identity of these bands is unknown. Immunoblotting failed to demonstrate factor VII as a contaminant of any of the high purity products, although factor VIIa could be detected in some lots of Immunine, Nanotiv, and 9MC by a clot-based assay. Factor X contaminated Aimafix, AlphaNine-SD, Factor IX VHP, Immunine, Nanotiv, and 9MC, but activation products of factor X were not detected.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7495064 TI - The utility of non-invasive tests for detection of previous proximal-vein thrombosis. AB - BACKGROUND: Despite the availability of several diagnostic methods for the detection of deep-vein thrombosis (DVT), the identification of previous episodes of DVT remains a diagnostic challenge. STUDY OBJECTIVE: To assess the reliability of a combination of a standardized clinical score with three non-invasive tests: compression ultrasonography (CUS), Doppler ultrasound (DUS), and photoplethysmography (PPG), in determining the presence or the absence of previous proximal DVT. METHODS: One hundred consecutive unselected outpatients were identified, who had undergone contrast venography six to nine years previously because of the clinical suspicion of DVT (confirmed in 43). They were blindly reinvestigated by a panel of trained operators unaware of venography results. They underwent a clinical evaluation of the lower limb, by applying a standardized score to five symptoms and six signs (grading each item from 0 to 3); a PPG test to determine the venous refilling time; a DUS test to determine the venous reflux separately in the common femoral and the popliteal vein; and a CUS test to determine vein compressibility in the same regions. RESULTS: An abnormal CUS test and/or the demonstration of venous reflux in the popliteal region and/or a high clinical score (> or = 8) identified twenty-four of the 43 (56%) DVT + patients with a specificity of 89%. The combination of normal CUS with the absence of venous reflux in both the common femoral and popliteal vein and a low clinical score excluded previous thrombosis in 45 (79%) of the 57 DVT patients (negative predictive value, 78%). Abnormal venous reflux in the isolated common femoral vein did not reliably predict the presence or absence of previous DVT. However, this occurred in only 13 (13%) patients. The PPG determination of venous refilling time did not improve the results obtained with the other tests. CONCLUSIONS: The combination of a standardized clinical evaluation with the results of CUS and DUS can reliably diagnose or exclude previous proximal-vein thrombosis in almost 90% of patients with previous episodes of suspected DVT. PMID- 7495065 TI - Varicose vein stripping--a prospective study of the thrombotic risk and the diagnostic significance of preoperative color coded duplex sonography. AB - Insufficiency of epifascial veins promotes venous ulceration and increases thromboembolic risk in general surgery patients. Epifascial varicose vein stripping is therefore considered the most effective prophylactic procedure. Thromboembolic risk of patients undergoing this surgical procedure has not yet been prospectively evaluated but appears to be lower than in general surgery patients. The gold standard of preoperative assessment of varicose surgery patients is ascending pressure phlebography, but this technique is invasive, time consuming and costly. We prospectively investigated 100 consecutive varicose vein surgery patients for postoperative thrombosis. Ascending pressure phlebography (APP) and colour coded duplex sonography (CCDS) were performed before and 10 to 21 days after the stripping operation in 100 and 70 patients, respectively. APP revealed no postoperative deep vein thrombosis in all 100 limbs investigated (0 percent; 95 percent confidence interval: 0 to 2.95). With regard to epifascial vein reflux there was good agreement between APP and CCDS (quadratic weighted kappa: 0.76). In 67 out of 73 superficial veins investigated excellent agreement of diagnostic accuracy was found for both diagnostic procedures (91.78 percent; 95 percent confidence interval: 82.96 to 96.92). We conclude that thrombotic risk of varicose vein surgery is low in properly selected patients. CCDS provides a high degree of accuracy in diagnosis of reflux and regular vein morphology and should therefore replace APP; however, APP does remain essential in the preoperative workup of atypical anatomical variants. PMID- 7495066 TI - Quantitative and qualitative analysis of platelet GPIb and von Willebrand factor in liver cirrhosis. AB - Numerous abnormalities of plasmatic coagulation and platelet function may contribute to the bleeding in liver cirrhosis with a defective platelet-von Willebrand factor interaction being a potential mechanism. To analyze GPIb and von Willebrand factor in cirrhosis, we quantified the number of GPIb molecules on the platelet surface by flow cytometry, assessed the total (and indirectly the internal) pool of GPIb by ELISA and measured the circulating amount of glycocalicin in plasma as a measure of proteolytic activity and platelet turnover. Von Willebrand factor was characterized by ELISA, by its ristocetin cofactor activity and by multimer analysis. Botrocetin-induced agglutination was used for functional analysis. The data from 8 well-characterized cirrhosis patients indicate that total GPIb is insignificantly increased to 46,000 +/- 5,000 molecules/P (normal: 39,500 +/- 2,000 [SEM]), surface-GPIb is normal with some variability and that the glycocalicin levels are 2-3 times higher than would be expected from the platelet count (= 100 +/- 5 x 10(9)/l). Von Willebrand factor antigen levels and activity were 400-500% of normal with a 22% reduction of the high molecular weight multimers. A significant hyperagglutination response to botrocetin was observed with platelets from both patients and controls using patient plasma as a source of von Willebrand factor. In conclusion, a hyperresponsiveness rather than a defective platelet-von Willebrand factor interaction can be observed in cirrhosis which may compensate for other hemostatic problems and appears to be mediated primarily by increased levels of von Willebrand factor. PMID- 7495067 TI - Diagnosis of subtype 2B von Willebrand disease in a patient with 2A phenotype of plasma von Willebrand factor. AB - Type 2A of von Willebrand disease refers to qualitative variants with decreased platelet dependent function that is associated with the absence of high molecular weight forms of von Willebrand factor (vWF) multimers. Type 2B refers to qualitative variants with increased affinity for platelet glycoprotein Ib. In this report we describe the study of a patient who has been previously diagnosed as having subtype 2A von Willebrand disease (vWD), because she had no heightened ristocetin-induced platelet aggregation, no large and intermediate molecular weight von Willebrand factor (vWF) multimers in plasma, and no increase in plasma vWF capacity to bind to normal platelets in the presence of low ristocetin concentrations. The DNA sequencing of the 3' part of the exon 28 of the vWF gene where most of the subtype 2A mutations have already been identified, did not detect any nucleotide change. At variance, a G to A transition changing the encoded amino acid residue from Val 553 to Met in mature vWF, was found in the 5' part of this exon. This mutation which has already been found in several unrelated families with 2B vWD and the increased binding of the patient platelet vWF on normal platelets in the presence of low ristocetin concentrations provide evidence for subtype 2B vWD. This study thus illustrates the importance of the molecular characterization of patients in the correct diagnosis and classification of type 2 vWD. PMID- 7495068 TI - Genetic determinants of plasma factor VII activity in the Japanese. AB - We investigated the frequency of the factor VII Gln353 (FVII Gln353) allele in Japan, where coronary artery disease is much less common and factor VII coagulant activity (FVIIc) is lower than in Western countries. Japanese males (n = 144) aged 40-59 years living in 2 different districts were studied, and the FVIIc and activated factor VII (FVIIa) levels were measured with human (FVIIa Hum) and bovine soluble tissue factor (FVIIa Bov). The frequency of the FVII Gln353 allele in the 2 districts was 0.026 and 0.052 (combined: 0.034), which was about one third of that in a European population. Thus, the lower FVIIc level in Japanese is not due to the effect of FVII Gln353 allele. Ten individuals with the Gln353 allele showed a 39% decrease of FVIIa Hum levels and a 29% decrease of the FVIIa Bov level compared with 134 individuals homozygous for the FVII Arg353 allele. These decreases of FVIIa were greater than the decrease of FVIIc (17%), suggesting that FVII Gln353 may be difficult to activate in vivo. The marked decrease of FVIIa levels in individuals with the FVII Gln353 allele suggests that they are genetically protected against cardiovascular disease irrespective of ethnic background, by virtue of their decreased FVIIa level. PMID- 7495069 TI - Genetic determination of coagulation factor VIIc levels among healthy middle-aged women. AB - A recent study (1) reported variation among men in clotting factor VIIc levels is associated with a genetic polymorphism detected by the restriction enzyme Msp I. The present study determined the Msp I genotype (Arg353, Gln353 alleles) for 189 women (mean age 53) who were subjects in the Healthy Women Study, a population study of CHD risk factor change at menopause. Women with the Arg/Arg genotype (n = 147) had an 16% higher (geometric) mean FVIIc level than those with the Arg/Gln (n = 41) genotype (1.21 vs 1.04 U/ml, p < 0.01), while the one subject with the Gln/Gln genotype had an FVIIc level of 1.00 U/ml. These results are consistent with those previously found in healthy men (1). In addition, women carrying the Gln allele did not exhibit the elevation in FVIIc with menopause and use of hormone therapy found among those with the Arg allele, suggesting that genotype may modify the observed rise in factor VIIc at menopause. Possibly because of the small sample size this interaction did not reach conventional levels of statistical significance. Results of multiple linear regression analyses controlling for age, hormone use, obesity, (ln) triglyceride levels, and family history of CHD found FVIIc levels to be significantly (p < 0.001) related to genotype. Thus, genotype appears to be a major determinant of FVIIc levels among women. PMID- 7495070 TI - Growth hormone deficiency impairs blood clotting and reduces factor VII coagulant activity in rat. AB - To investigate pituitary effects on the vitamin K-dependent coagulation factors, female rats were hypophysectomized (hypox) and treated with growth hormone (GH), cortisone, thyroxine, vitamin K, or saline. After 11 days of treatment, the prothrombin time, platelet count, and factors II, VII, IX, and X were determined. The prothrombin time was 52.9 +/- 1.2% for control rats and 39.1 +/- 0.8% for hypox rats (mean +/- SEM; p < 0.001). All factors decreased after hypophysectomy, reaching significance for factor VII (from 264 +/- 23% to 131 +/- 9%; p < 0.001) and factor IX (from 28.4 +/- 2.2% to 17.1 +/- 2.5%; p < 0.01) while the platelet count was unaffected. When hypox rats were treated with GH, the prothrombin time increased to 50.9 +/- 1.0% (p < 0.001) and factor VII to 299 +/- 10% (p < 0.001). Factor II, IX, and X were slightly increased after GH substitution but not after cortisone, thyroxine, or vitamin K treatment. To summarize, GH is of importance for normal hemostasis in the female rat. PMID- 7495071 TI - Comparison of the pharmacokinetic profiles of three low molecular mass heparins- dalteparin, enoxaparin and nadroparin--administered subcutaneously in healthy volunteers (doses for prevention of thromboembolism). AB - The present trial was designed to comparatively investigate the pharmacokinetic profile and evaluate the apparent bioavailability pattern of three already marketed low molecular mass heparins (LMMHs): dalteparin (Fragmin), nadroparin (Fraxiparin), and enoxaparin (Lovenox) given by subcutaneous route. The study was carried out in 20 healthy young volunteers given, according to a cross over design, a single subcutaneous injection of the doses recommended for the prophylaxis of deep vein thrombosis (commercial preparations, prefilled syringes): dalteparin 2,500 IU (= 2,500 IU anti-Xa), nadroparin 7,500 ICU (= 3,075 IU anti-Xa), enoxaparin 20 mg (= 2,000 IU anti-Xa) and enoxaparin 40 mg (= 4,000 IU anti-Xa). Of the markers used, activated partial thromboplastin time (APTT), thrombin clotting time (TCT), Heptest, anti-thrombin (aIIa) activity and anti-Xa (aXa) activity, the most pertinent parameter (from a biodynamic viewpoint) is plasma aXa activity. We demonstrated that dalteparin, nadroparin and enoxaparin exhibit statistically significantly different pharmacokinetic and overall disposition patterns. Normalized to the same injected dose (1,000 IU aXa), the relative actual amount of plasma anti-Xa activity generated by enoxaparin is 1.48 times greater (p < 0.001) than that of nadroparin and 2.28 times greater (p < 0.001) than that of dalteparin while the plasma amount induced by nadroparin is 1.54 times greater (p < 0.001) than that of dalteparin. The apparent total body clearance of enoxaparin doses (CL/F = 16.7 +/- 5.5 and 13.8 +/- 3.2 ml/min) is significantly smaller than those of nadroparin (CL/F = 21.4 +/ 7.0 ml/min; p < 0.01) and dalteparin (CL/F = 33.3 +/- 11.8 ml/min; p < 0.001) while dalteparin apparent clearance is about 1.5-fold greater (p < 0.001) than that of nadroparin. These LMMHs also differ by their renal excretion pattern: more fragments exhibiting an anti-Xa activity are recovered in urine following enoxaparin doses (6.4 and 8.7% of the dose, respectively) than following nadroparin (3.9%) and dalteparin (3.4%) injection. These differences in the disposition profiles explain why the apparent elimination half life t1/2 values of the LMMHs compared here are different: dalteparin: 2.8 h; nadroparin: 3.7 h; and enoxaparin: 4.1 h. Whether or not these differences may contribute to explain the different safety/efficacy balance of each of these antithrombotic medications remains to be discussed and needs further studies. PMID- 7495072 TI - Antithrombotic activity in vivo of SDZ 217-766, a low-molecular weight thrombin inhibitor in comparison to heparin. AB - Antithrombotic potency of SDZ 217-766, a potent inhibitor of thrombin and other trypsin-like serine proteases, was studied in comparison with heparin in rat models of thrombin induced lung platelet accumulation, of thrombosis in arterio venous shunt, and of venous thrombosis induced by tissue factor. Thrombin-induced platelet accumulation in the lung was inhibited dose-dependently by SDZ 217-766 following intravenous (i.v.) administration of 0.03 mg/kg to 0.3 mg/kg as well as by intraduodenal (i.d.) administration of 3 mg/kg and 10 mg/kg. Comparable inhibitory effects were observed with heparin at 30 IU/kg and 100 IU/kg. In the rat arterio-venous shunt, following i.v. administration of SDZ 217-766, thrombus formation was inhibited by 40% at 0.1 mg/kg, by 60% at 0.3 mg/kg and was abolished at 1.0 mg/kg whilst APTT was prolonged 1.1 fold over the control value at 0.1 mg/kg and 2.7 fold at 1.0 mg/kg. Similar inhibitory effects were observed following i.d. administration of 10 and 30 mg/kg with only marginal (1.2 to 1.8 fold) APTT elevation. In the same model, heparin administered either i.v., 30-300 IU/kg, or subcutaneously, 100 and 300 IU/kg, inhibited thrombus formation dose dependently but in contrast to SDZ 217-766, the inhibitory effect was paralleled by 5-to > 10 fold APTT elevation over baseline. In the venous thrombosis model, SDZ 217-766 infused at 10 micrograms/kg/min and 20 micrograms/kg/min, reduced thrombus formation by 35% and 70%, respectively. In comparison, thrombus formation was decreased by 22% when heparin was infused at 1 IU/kg/min, and abolished at 3 IU/kg/min.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7495073 TI - Inhibition of prothrombinase by antithrombin-heparin at a macroscopic surface. AB - The antithrombin-dependent inhibition of prothrombinase, assembled at a macroscopic surface, was studied under flow conditions utilizing a tubular flow reactor that consists of a phospholipid-coated glass capillary. Prothrombinase activity was determined from steady-state rates of thrombin production upon perfusion with prothrombin and from factor Va-associated factor Xa activity present in the flow reactor. The prothrombinase density was maintained at a low level (0.03 fmol/cm2) to assure that the rate of thrombin production reflected the amount of prothrombinase present in the capillary. Perfusion of the flow reactor with antithrombin resulted in an exponential decrease of prothrombinase activity in time. The second order rate constant (8.5 x 10(4) M-1min-1) is comparable with the rate of inactivation of free factor Xa. Inhibition was much faster when antithrombin was complexed with heparin. The second order rate constants of inhibition decreased with decreasing heparin chain length: 9.6 x 10(7), 4.5 x 10(7) and 0.39 x 10(7) M-1min-1 for unfractionated heparin, low molecular weight heparin and synthetic pentasaccharide heparin, respectively. In the presence of prothrombin (0.2 microM), however, the heparin-dependent rate of inhibition of prothrombinase was about 50-fold lower. The heparin-independent inhibition of prothrombinase by antithrombin (4 microM) in the presence of prothrombin (0.2 microM) was virtually negligible. At a 70-fold higher surface density of prothrombinase (2 fmol/cm2) prothrombinase activity was much faster inactivated. The rate of thrombin production, however, was not affected. In conclusion, at low prothrombinase densities, prothrombin efficiently protects prothrombinase from inhibition.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7495074 TI - Cloning of an avian antithrombin: developmental and hormonal regulation of expression. AB - We present the complete amino acid sequence of a non-mammalian antithrombin together with a structural comparison to its mammalian homologs. In the majority of cases, but not all, the analyses indicate high degrees of primary structural identity within regions of predicted or demonstrated functional importance. We have also taken advantage of the avian model to define the ontogeny of antithrombin mRNA expression. Northern analyses indicate chicken antithrombin mRNA levels to be detectable in the liver early during development and increase to reach adult levels just prior to hatching. Although no sex-dependent differences in antithrombin mRNA levels were detected, the administration of estrogen to male roosters was observed to cause a 3-fold decrease in antithrombin mRNA levels, suggesting that the hormone may alter circulating antithrombin levels via a pre-translational mechanism. PMID- 7495075 TI - Calcium ion-dependent monoclonal antibody against human fibrinogen: preparation, characterization, and application to fibrinogen purification. AB - We have produced a high-affinity monoclonal antibody classified as IgG1 with kappa-type light chains that recognizes the calcium ion(Ca2+)-dependent conformation of the D-domain of human fibrinogen. Binding of fibrinogen in solution to the insolubilized antibody increased in the presence of increasing concentrations of up to 2 mM Ca2+, the half-maximal binding being reached at 130 microM Ca2+. The dissociation constant was estimated to be 1.6 x 10(-8) M at 2 mM Ca2+. The antibody was found also to be dependent on other divalent metal ions including Zn2+, Mn2+, Co2+ and Cu2+, but not Ba2+, Mg2+ or Sr2+. The synthetic Gly-Pro-Arg-Pro-amide peptide, which has recently been shown to bind to close proximity to the calcium binding site in the D-domain, was unable to elicit the conformation for the antigen to be recognized by this antibody. This antibody was found to be a suitable ligand for the immunoaffinity chromatography of normal and abnormal fibrinogens directly from citrated plasma depleted of the vitamin K dependent proteins or heparinized plasma by eliminating the precipitation procedure widely adopted in conventional techniques of fibrinogen purification. Indeed, fibrinogen Marburg I with the A alpha chains depleted of the carboxy terminal A alpha(461-610) residue segment has been purified by this technique, although this dysfibrinogen was difficult to purify by conventional precipitation techniques. PMID- 7495076 TI - Differences in the interactions of lupus anticoagulant IgG with human prothrombin and bovine prothrombin. AB - Lupus anticoagulant (LA) IgGs have been reported to inhibit more effectively and consistently the Xa/Va/phospholipid complex-catalyzed activation of human prothrombin that the Xa/Va/phospholipid complex-catalyzed activation of bovine prothrombin. This led us to carry out studies to determine whether the ability to inhibit the activation of prothrombin of LA IgGs, separated from the plasma of 15 patients by protein A affinity chromatography, could be related to the ability of the LA IgGs to bind to prothrombin under various experimental conditions. Of 14 LA IgG preparations tested all prolonged to a variable but substantial extent the dilute Russell's viper venom time (dRVVT) of human plasma but only minimally prolonged the dRVVT of bovine plasma. In a purified prothrombin activation system with a rate limiting concentration of phospholipid, all 15 LA IgG preparations inhibited the activation of human prothrombin with the majority showing > 50% of inhibition. In contrast, only one LA IgG markedly inhibited (> 50%) the activation of bovine prothrombin and five others moderately inhibited (25-40%) the activation of bovine prothrombin. Nevertheless, the majority of LA IgG preparations bound to immobilized bovine prothrombin on a Western blot and also to immobilized bovine prothrombin on a microtiter well. In an ELISA in which phosphatidylserine (PS) was immobilized on microtiter wells, bovine prothrombin supported the binding of 10 of 15 LA IgG preparations to PS. However, the extent of binding was lower than that observed with human prothrombin.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7495077 TI - Measurement of activated factor IX in factor IX concentrates: correlation with in vivo thrombogenicity. AB - Current in vitro tests for thrombogenicity of FIX concentrates used for prothrombin complex concentrates (PCCs), are of little value when applied to high purity FIX (HP FIXs). In the present study, we have developed a chromogenic assay for activated FIX (FIXa) and evaluated its ability to predict in vivo thrombogenic potential of HP FIXs in a modified Wessler stasis model. Among the HP FIXs, only 1 out of 7 products had no detectable FIXa; this product also showed no in vivo thrombogenicity. In the other 6 products, FIXa content ranged from 0.15-1.2 U/1000 in FIX, and all showed some evidence of in vivo thrombogenicity, with mean thrombus scores ranging from 0.25-4. There was a significant positive correlation (r = 0.55, p < 0.02) between FIXa levels and in vivo thrombogenicity of HP FIXs. NAPTT data were not significantly correlated with the in vivo results and the TFCT also showed no direct correlation with the mean thrombus score. These results indicate that HP FIXs may still carry a small residual thrombotic risk and measurement of FIXa content of these products may be a better predictor of thrombogenicity than the current in vitro tests. PMID- 7495078 TI - Therapeutic advantage of recombinant human plasminogen activator in endocarditis: evidence from experiments in rabbits. AB - In infective endocarditis vegetations are stabilized by fibrin. To learn if fibrin digestion would be therapeutic, experimental endocarditis was induced in rabbits by inoculation with a platelet-aggregating strain (Agg+) of Streptococcus sanguis and treated with recombinant tissue plasminogen activator (rt-PA), rt-PA with penicillin, or penicillin alone. Control rabbits were inoculated with saline. All treatments of Agg+ endocarditis reduced the mass of valvular vegetations and clinical signs of endocarditis, including the frequency of left axis deviation and heart ischemia. rt-PA with penicillin was more effective than penicillin or rt-PA alone, reducing the mass of vegetations and clinical signs to that of saline controls. Within 50 min, rt-PA cleared 5-fold more 111Indium labelled platelets from the heart than untreated rabbits and 1.4-fold more after 3 days. Combined with penicillin, thrombolytic therapy for human endocarditis should be reconsidered. PMID- 7495079 TI - Comparison of antithrombotic effects of GPIIb-IIIa receptor antagonist and TXA2 receptor antagonist in the guinea-pig thrombosis model: possible role of TXA2 in reocclusion after thrombolysis. AB - The effects of Ro 44-9883, a new specific antagonist of platelet glycoprotein IIb IIIa receptor, on thrombus formation and reocclusion after thrombolysis induced by tissue-type plasminogen activator (t-PA) were compared with those of vapiprost, a thromboxane (TX) A2 receptor antagonist, using a photochemically induced thrombosis model in the guinea-pig femoral artery. Pretreatment with Ro 44-9883 (5, 10 and 20 micrograms/kg/min, i.v.) prolonged the time required to occlude the artery in a dose-dependent manner. Ro 44-9883 at 10 and 20 micrograms/kg/min significantly inhibited ex vivo platelet aggregation in whole blood induced by collagen, ADP or U46619. Vapiprost 0.3 mg/kg inhibited thrombus formation and platelet aggregation induced by collagen or U46619, to the same extent as Ro 44-9883 at the higher doses. In the thrombolysis study, Ro 44-9883 at the higher doses given as comedication with t-PA reduced the time to achieve reperfusion and increased the vascular patency after successful reperfusion. Vapiprost also significantly reduced the time to reperfusion and prevented reocclusion. However, the vascular patency after thrombolysis by t-PA with vapiprost was significantly increased compared with Ro 44-9883. Ro 44-9883 inhibited platelet aggregation, but did not prevent TXA2 formation in platelets. Thus, vascular contraction mediated by platelet-derived TXA2 may be responsible for lower efficacy of Ro 44-9883 against reocclusion compared with vapiprost. These results indicate that not only platelet aggregation but also vasoconstriction may contribute to reocclusion after t-PA-induced thrombolysis in the guinea-pig. PMID- 7495080 TI - Ristocetin-induced platelet agglutination stimulates GPIIb/IIIa-dependent calcium influx. AB - We found that intracellular Ca2+ concentration ([Ca2+]i) increased during ristocetin-induced agglutination of aequorin loaded platelets resuspended in plasma. Chelation of extracellular Ca2+ had no effect on platelet clumping, but delayed and greatly reduced Ca2+ increase, indicating that it derived for the most part from Ca2+ influx. Nine monoclonal antibodies (MA) against glycoprotein (GP) Ib largely prevented ristocetin-induced platelet clumping and [Ca2+]i increase, while three anti-GPIb MA with no effect on platelet clumping did not interfere with Ca2+ movement. In unstirred samples platelet agglutination was greatly reduced and [Ca2+]i increase was abolished, suggesting that close platelet-to-platelet contact, in addition to von Willebrand factor (vWF) binding to GPIb, is necessary for Ca2+ transient. Nine MA against GPIIb/IIIa, the gly-arg gly-asp-ser (GRGDS) peptide and GPIIb/IIIa complex dissociation had no effect on platelet agglutination, but significantly reduced Ca2+ increase. Our results suggest that platelet clumping induced by vWF binding to GPIb is responsible for GPIIb-IIIa dependent Ca2+ influx. PMID- 7495081 TI - Exposure of human platelets to plasmin results in the expression of irreversibly active fibrinogen receptors. AB - Although plasmin can trigger strong platelet responses such as shape change and exocytosis of internal granules, limited platelet aggregation is induced by this proteinase, owing to its capacity to rapidly proteolyse secreted adhesive proteins. In this context, we have investigated the state of activation of the fibrinogen receptor, the integrin alpha IIb beta 3, on platelets exposed to plasmin. Following incubation with plasmin at 37 degrees C, washing, and resuspension, platelets exhibit a moderate, low-velocity aggregation when stirred in the presence of fibrinogen. Optimum aggregability is observed when platelets have been exposed to plasmin activity of approximately 0.5 CU/ml for 20 min, and aggregation is insensitive to the presence of antagonists such as prostaglandin (PG) E1 and apyrase. Plasmin-induced platelet aggregability is associated with the expression of active fibrinogen receptors on the cell surface, which, using a 125I-fibrinogen binding assay, can be quantified to approximately 2,300 molecules per platelet. Exposure of active alpha IIb beta 3 receptors appears to depend partially, but not totally on a metabolic activation and granule exocytosis at the time of incubation with plasmin. In contrast with alpha-thrombin, plasmin induced activation of alpha IIb beta 3 is sustained and cannot be reversed by exposure of platelets to PGE1. Immunoblotting analysis of the receptor subunits shows no extensive proteolytic modification of alpha IIb beta 3 by plasmin, and only reveals a limited proteolysis of the aminoterminal domain of the alpha IIb subunit. In addition to their capacity to aggregate in the presence of fibrinogen alone, plasmin-treated platelets also show a potentiated aggregability in response to low doses of ADP.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7495082 TI - Transformation of cellular factor XIII into an active zymogen transglutaminase in thrombin-stimulated platelets. AB - The cellular form of blood coagulation factor XIII (FXIII) is present in platelets, monocytes and macrophages. During long-term stimulation of platelets by thrombin cellular FXIII becomes activated and cross-links proteins, however, the mechanism of its activation has not been elucidated. It was shown that, contrary to plasma FXIII, the intracellular activation of platelet FXIII does not involve proteolysis. FXIII remained intact in thrombin-activated platelets, i.e., the activation peptide was not removed from the molecule. Part of the zymogen FXIII molecules, however, assumed an active configuration as was demonstrated both by the measurement of transglutaminase activity and by active-site-SH titration. These findings clearly indicate that during platelet activation, when intracellular Ca2+ concentration is raised, a slow non-proteolytic transformation of FXIII zymogen into an active transglutaminase occurs. PMID- 7495084 TI - Cultured endothelial cells regulate platelet adhesion to their extracellular matrix by regulating its von Willebrand factor content. AB - Endothelial cells and their extracellular matrix formed in vitro are often used as a model for subendothelium in studies on platelet-vessel wall interaction. We have characterized the influence of culture conditions of endothelial cells on the formation of extracellular matrix and on the interaction of the matrix with platelets. Passage number, time of confluence, serum concentration and the addition of heparin, growth factors and antibiotics to the culture medium were varied and the extracellular matrices were isolated. The amount of fibronectin and von Willebrand factor present in the matrix were measured and the number of platelets adhering to these matrices after perfusion with citrated whole blood at a shear rate of 1000 s-1 was determined. A three times increase of the amount of von Willebrand factor in the matrix was found when the serum concentration was increased from 2.5% to 30%. When the passage number of the cells was increased or the period during which the cells were at confluence was extended, the amount of von Willebrand factor in the matrix was decreased up to 50%. Addition of heparin or ECGS (endothelial cell growth supplement) decreased the von Willebrand factor content in the matrix. Addition of penicillin or streptomycin to the culture medium had no influence on the amount of von Willebrand factor deposited in the matrix or secreted into the medium, however, other antibiotics such as gentamycin and neomycin decrease the amount of von Willebrand factor in the matrix. No influence on the amount of fibronectin in the matrix was found under all conditions tested.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7495083 TI - Augmentation of vascular endothelial barrier function by heparin and low molecular weight heparin. AB - Glycosaminoglycans (GAGs) are an important component of endothelial barrier function. Early passage human umbilical vein endothelial cells were grown to confluence on transparent micropore filters and barrier function assessed as transendothelial electrical resistance (TEER) and permeability to albumin and sucrose. Unfractionated heparin and the LMW heparin Clexane decreased endothelial permeability to both sucrose and albumin and increased TEER. Chondroitin 6 sulphate also augmented barrier function, but other GAGs had no effect. Interleukin-1 increased permeability to albumin and sucrose and decreased TEER. Although heparin attenuated the effect of IL-1 on TEER and sucrose permeability, it could not restore the barrier to albumin transfer. Denuded endothelial matrix presented a negligible barrier, which was not enhanced by heparin. When sulphation of endogenous GAGs was inhibited by chlorate, barrier function was compromised and was not restored by exogenous heparin. Thus heparin enhances the barrier function of resting endothelium, but cannot completely overcome the increased permeability resulting from exposure to IL-1 or substitute for endogenous GAGs. PMID- 7495085 TI - The suppression of the coagulation of nonanticoagulated whole blood in vitro by human umbilical endothelial cells cultivated on microcarriers is not dependent on protein C activation. AB - Human umbilical vein endothelial cells (HUVEC) were cultivated on globular microcarriers in order to improve the endothelial cell surface to blood-volume ratio over the conventional flat bed cultures. HUVEC-beads were tested for their modulation of blood coagulation using a combination of two steps: HUVEC-beads were added into the syringe used for the venipuncture, in order to achieve immediate contact between cells and blood, and no anticoagulant was used during the incubation time of HUVEC-beads with whole blood. The coagulation initiation produced by venipuncture was almost completely suppressed as judged by thrombin measurements over a period of 60 min. The activated partial thromboplastin time showed a prolongation by a factor > 3. Direct measurements of activated protein C (APC) were negative. Moreover, inhibition of APC-generation with a monoclonal anti-human protein C antibody did not affect the anticoagulant properties of endothelial cells. Therefore the anticoagulant properties exerted by HUVEC-beads are not dependent on APC. PMID- 7495086 TI - Terminology for macromolecular derivatives of crosslinked fibrin. On behalf of the Subcommittee on Fibrinogen of the Scientific and Standardization Committee of the ISTH. PMID- 7495087 TI - Prophylaxis of deep vein thrombosis in orthopaedic surgery. PMID- 7495088 TI - The usefulness of D-Dimer in the diagnosis of postoperative DVT. PMID- 7495089 TI - Kinetic behaviour of the TFPI and anti-FXa effects, after the injection of a LMWH (RO-11) in healthy subjects. PMID- 7495090 TI - Influence of factor V and factor Va on APC-induced cleavage of human factor VIII. PMID- 7495091 TI - Apparent heterozygous type II protein C deficiency caused by the factor V 506 Arg to Gln mutation. PMID- 7495092 TI - Diagnosis of APC resistance in patients on oral anticoagulant therapy. PMID- 7495093 TI - Evaluation of APC resistance in the plasma of patients with Q506 mutation of factor V (factor V Leiden) and treated by oral anticoagulants. PMID- 7495094 TI - Continuous infusion of prostacyclin decreases plasma levels of t-PA and PAI-1 in primary pulmonary hypertension. PMID- 7495095 TI - Hepatitis C and pasteurised factor VIII and IX concentrates. PMID- 7495096 TI - Preliminary in vivo evaluation of a nanofiltered factor IX concentrate. PMID- 7495097 TI - Regulation of the extrinsic pathway system in health and disease: the role of factor VIIa and tissue factor pathway inhibitor. PMID- 7495098 TI - Experimental thrombosis model induced by free radicals. Application to aspirin and other different substances. AB - A large number of experimental studies suggests that oxygen free radicals play a major role in the pathogenesis of the myocardial lesions observed during the sequence ischemia-reperfusion. The purpose of this study was to determine whether oxygen free radicals can induce thrombosis. In so doing we have developed a new experimental thrombosis model. Reproducible focal thrombosis has been achieved by irradiating mesenteric arterioles of rat for variable time with green filtered light issuing from a mercury lamp after systemic injection of different rose bengal doses. The number of emboli that remove in the blood (N), the duration of total occlusion (T) and the number of emboli per minute were then measured. As control, no rose bengal administration was done and the vessels were exposed to the filtered light. In comparison with this control, results clearly showed that free radicals always induced thrombosis and the induced thrombus was mainly composed of platelets. In this new thrombosis model induced by free radicals antithrombotic drugs (aspirin, 200 mg/Kg, heparin, 2 mg/Kg) and antioxidants (vitamin C, 10 and 20 mg/Kg, allopurinol, 200 and 300 mg/Kg, vitamin E, 500 and 1000 mg/Kg) have been tested. Results have shown that only heparin and vitamin E had an antithrombotic effect on thrombus formation induced by free radicals. This model should be useful in studying the effects of different drugs and could lead to new treatment modalities for ischemic accident and other cardiovascular diseases. PMID- 7495099 TI - Treatment with recombinant human erythropoietin induces a moderate rise in hematocrit and thrombin antithrombin in healthy subjects. AB - Recombinant human erythropoietin (EPO) therapy in uremic patients raises the hematocrit (Hct) and increases physical exercise capacity (1,2) and quality of life (1). In general, partial correction of anemia to subnormal levels in uremic patients has proven to be safe with few serious adverse effects apart from hypertension (3). Ever since the advent of EPO the prospect of abuse of the hormone by sportsmen has been subject to scrutiny. Both maximal oxygen uptake and endurance capacity are increased after EPO treatment in healthy subjects (4). Moreover, EPO treatment in healthy subjects has been found to induce an accentuated blood pressure reaction after submaximal exercise (5). Previous studies have shown that extreme physical exertion can predispose to an increased intravascular coagulation (6). Moreover there is a significantly increased risk of thrombosis in patients with myeloproliferative disorders, particularly in polycythemia vera (7). An enhanced risk of cardiovascular events may therefore arise should sportsmen abuse EPO as a blood doping agent. The aim of this study was to examine the effects of an EPO-induced increase in Hct on the coagulation system in healthy subjects. PMID- 7495100 TI - Dextran excretion rate in desmopressin treated volunteers. PMID- 7495101 TI - Detection of lupus anticoagulant: an evaluation of routines for preparation and storage of plasma. PMID- 7495102 TI - A rapid and simple isolation of endothelial cell growth supplement from mouse brain. PMID- 7495103 TI - Protein Z deficiency: a new cause of bleeding tendency. AB - Protein Z is a vitamin K-dependent plasma protein synthesized by the liver. Protein Z promotes the association of thrombin with phospholipid surfaces. So far, nothing is known about the clinical relevance of protein Z except alterations measured in patients under oral anticoagulant treatment. We propose that in protein Z deficiency a bleeding tendency might result because of the interaction of protein Z with thrombin on phospholipid endothelial surfaces. Therefore, we examined 36 patients with bleeding tendency of unknown origin, who were not under oral anticoagulant treatment and had normal liver synthesis function. Mean protein Z value of the patients with bleeding tendency was significantly diminished in comparison to the healthy control group. Twenty-one of the bleeding patients had lower protein Z levels than the lowest protein Z level of the control group. In conclusion, protein Z deficiency is described as a new type of bleeding tendency. PMID- 7495104 TI - Altered lysis resistance of platelet-rich clots in patients with insulin dependent diabetes mellitus. AB - The fibrinolytic resistance of platelet-rich arterial thrombi received much attention. Clot lysis method was used to assess the in vitro fibrinolytic properties in diabetes mellitus. Platelet rich (PRP) clots were formed by addition of thrombin, and lysis was induced by tissue-plasminogen-activator. The coagulation and lysis was followed by the light scattering properties. A special pattern of good initial lysis followed by a second clotting phase was observed in more than half of insulin dependent diabetic patients, while a similar pattern of clot-lysis was only occasionally found in non-insulin dependent diabetes mellitus or in the healthy control group. Following the thrombin activation of washed, gel filtered platelets, the supernatants possessed an inhibitory action on in vitro lysis of PPP-clots. This suppression was remarkably stronger in IDDM, along with the highest PAI-1 activity concentration ratio of the platelet lysates, compared to plasmatic levels. The relation of this special type of PRP clot-lysis resistance to diabetic vascular complications needs further clarifying and investigations. PMID- 7495105 TI - Detection of monocyte tissue factor after endotoxin stimulation: comparison of one functional and three immunological methods. AB - Monocyte procoagulant activity is mainly due to tissue factor (TF) expression, but functional assays may not be sufficiently accurate in clinical use, making useful a determination of TF antigen level. The aim of this study was to compare the results of one functional and three immunological TF assays (ELISA, immunocytochemical staining on slides and flow immuno cytometric analysis), in normal monocytes, after standardized stimulation by endotoxin. TF expression was determined in blood mononuclear cells isolated by gradient centrifugation and cultured, with or without various concentrations of endotoxin. On lysed cells, TF activity was determined by amidolytic assay and TF antigen level was determined, after triton extraction, by ELISA (Imubind, American Diagnostica). Mouse monoclonal antibody against TF (4508, American Diagnostica) was used for 1) immunocytochemical (ICC) staining on cytocentrifuge slides (Avidine-Biotine peroxidase-Complex revelation) and 2) flow cytometric analysis using indirect labeling (Fab'2 Fluoresceine Isothyocyanate revelation). The determination of TF activity and TF antigen by ELISA method were equally sensitive to low concentration of endotoxin (0.005 EU/ml) and well correlated in the presence of higher concentrations of endotoxin. ICC led to a qualitative detection with a similar sensitivity to endotoxin stimulation. Flow cytometric analysis was poorly sensitive to increasing stimulation of monocytes. Of note, the functional, ELISA and immunocytochemical assays for monocyte TF expression were sensitive to endotoxin concentrations as low as 0.005 EU/ml. PMID- 7495106 TI - Flurbinitroxybutylester: a novel anti-inflammatory drug has enhanced antithrombotic activity. AB - We have recently shown that the introduction of a nitroxybutylester moiety into flurbiprofen, to form Flurbi-NO, results in a compound with markedly reduced undesired effects in the gastrointestinal tract. This effect has been shown to be linked to nitric oxide release from the Flurbi-NO. Here we have investigated whether this is associated with a reduction in platelet aggregability in vivo, as assessed in a mouse model of thromboembolism and a rat model of platelet aggregation, and found in both models that Flurbi-NO is more potent than flurbiprofen at inhibiting collagen-induced platelet aggregation. Further in vitro studies using human washed platelets and cells in culture suggest that this is due to the release of NO from Flurbi-NO following the action of (possibly plasma) esterases. Together with our earlier data, these results strongly suggest that Flurbi-NO and other members of this class of drugs, have particular potential as anti-thrombotic agents devoid of gastrointestinal side effects. PMID- 7495107 TI - Activation of the protein C pathway in acute sepsis. AB - To obtain quantitative information on the in vivo activation of the protein C system during the acute phase of sepsis, several components of the protein C pathway were studied in 18 patients. Blood samples were obtained one day after diagnosis (day 1) and, in 11 patients, also on the fourth and tenth days after diagnosis (days 4 and 10). On day 1, patients showed laboratory signs of haemostatic alterations such as positive fibrinogen/fibrin degradation products, and increased thrombin:antithrombin-III (TAT) complex levels. Compared with the control group, patients on day 1 had significantly decreased (p < 0.001) antigenic protein C (69 +/- 28%) and protein C inhibitor (PCI) (33 +/- 22%) whereas a significant increase in the levels of activated protein C (APC) complexed with alpha 1-antitrypsin (alpha 1AT) (APC:alpha 1AT, 26 +/- 15 ng/mL) and APC:PCI complex (3.0 +/- 2.0 ng/mL), and in the level of plasma kallikrein (KK) complexes with PCI (KK:PCI) (31 +/- 22 ng/mL) was observed. There was a positive correlation between APC:alpha 1AT and TAT complex levels (r = 0.597, p = 0.009). In the follow-up a trend toward normal values in antigenic protein C and PCI, and in APC:PCI and KK:PCI complex levels was found. However, PCI remained significantly decreased compared to normal values. C4b-binding protein, alpha 1AT, and TAT and APC:alpha 1AT complexes did not show any significant variations during the course of the disease, suggesting the contribution of the inflammatory and haemostatic responses, in spite of the good recovery of the patients. This study shows that in the course of sepsis, patients experience a generalized activation of the protein C pathway which was more prominent on day 1, resulting in the consumption of protein C and PCI and in the increase of APC:inhibitor complexes. Moreover, these data provide further evidence that KK:PCI circulating complexes occur in vivo. PMID- 7495108 TI - Beneficial effect of CV-4151 (Isbogrel), a thromboxane A2 synthase inhibitor, in a rat middle cerebral artery thrombosis model. AB - Effects of thromboxane A2 (TXA2) synthase inhibitors (CV-4151 and ozagrel) on cerebral thrombosis and cerebral damage were examined in a rat middle cerebral artery (MCA) thrombosis model and their potencies were compared with the conventional antithrombotic agents, aspirin and ticlopidine. CV-4151 significantly inhibited photochemically induced MCA thrombosis by oral (1 and 10 mg/kg) and intravenous (1 mg/kg) administration. Ozagrel (10 mg/kg, p.o.) also inhibited it. The potency of CV-4151 was about 10 times stronger than that of ozagrel, being comparable with the inhibition of blood TXA2 generation. Aspirin (100 mg/kg, p.o.) and ticlopidine (300 mg/kg, p.o.) showed an inhibitory tendency on MCA thrombosis. Twenty-four h after photochemical stimulation, cerebral edema and cerebral infarction were observed, and the lactate content in the brain increased. CV-4151 and ozagrel prevented this edema, and the antiedema effects of the drugs were correlated with the antithrombotic effect on thrombotic MCA occlusion. CV-4151 (10 mg/kg, p.o.), furthermore, significantly reduced the infarct size and inhibited the increase in lactate content. These results indicate that TXA2 synthase inhibitors inhibit cerebral damage by inhibition of MCA occlusion with thrombosis, probably resulting from the inhibition of TXA2 generation, and their effects are superior to those of aspirin and ticlopidine. TXA2 might play an important role in cerebral damage in the MCA thrombosis model. CV-4151 might be a useful drug for the treatment of cerebral thrombosis and for the prevention of cerebral infarction. PMID- 7495109 TI - Should we treat localized prostate cancer? An opinion. AB - Localized prostate cancer is a progressive disease if left untreated. However, cancer-specific mortality is low in patients with moderately and well differentiated prostate cancer treated with observation and delayed hormonal therapy, being 13% at 10 years and 20% to 30% at 15 years. By and large, radiation therapy does not appear to improve survival in these patients. With modern surgical techniques, mortality from prostate cancer is lowered by 23% to 65% in patients with moderately or well-differentiated tumors. However, the impact on relative cancer-specific survival is modest, since the mortality rate in untreated patients is low. The survival of conservatively managed patients with poorly differentiated prostate cancer is dismal: here radiation therapy or surgery significantly improves outcome. The QOL of patients with localized prostate cancer is significantly affected by the occurrence of distant metastasis. Metastatic rates are high in patients who are followed with observation and delayed endocrine treatment (19% to 85%). We were unable to deduce the effects of radiation therapy on grade-specific metastatic rates at 10 and 15 years. The only surgical series that addresses the issue shows a 50% to 80% reduction in metastatic rates. This results in an improvement in metastasis free survival of 19% to 300%. The reduction in metastatic rates with surgery holds true for patients with poorly, moderately, or well-differentiated tumors. However, a significant proportion of the surgical patients were treated with adjuvant endocrine therapy, and it is impossible to identify the benefit from surgery and the benefit from adjuvant therapy. Radical prostatectomy improves survival in men who are 65 years or younger with moderately or well differentiated adenocarcinoma of the prostate, and in men 75 years or younger who have poorly differentiated adenocarcinoma of the prostate. Its efficacy in reducing cancer-specific mortality in patients who have a survival expectancy of less than 15 years (older than 65 years) and moderately or well-differentiated adenocarcinoma of the prostate is less clear. Radical prostatectomy, with or without adjuvant hormonal therapy, decreases metastatic rates in men with a life expectancy of 10 years or more (age 75 years or younger) irrespective of tumor grade and, thus, should improve the QOL expectancy in these men. Nevertheless, between 20% and 60% of patients undergoing radical prostatectomy have biochemical recurrence, as defined by a detectable PSA, at 10 years of follow-up. This is worrisome and may portend clinical failure with longer follow-up.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 7495110 TI - Small cell carcinoma of the bladder and prostate. PMID- 7495111 TI - Can finasteride reverse the progress of benign prostatic hyperplasia? A two-year placebo-controlled study. The Scandinavian BPH Study Group. AB - OBJECTIVES: To study if placebo-induced improvement in men with symptomatic benign prostatic hyperplasia (BPH) is maintained over 2 years, and to study the efficacy and safety from intervention with finasteride 5 mg for 24 months. METHODS: This was a multicenter, double-blind, placebo-controlled study involving 707 patients with moderate symptoms of BPH enrolled at 59 centers in five Scandinavian countries. Following enrollment and a 4-week single-blind placebo run-in period, patients were randomized to receive finasteride 5 mg once daily or placebo for 24 months. Urinary symptoms, urinary flow rate, prostate volume, postvoiding residual urinary volume, and serum concentrations of prostate specific antigen together with laboratory safety parameters were measured at entry and at months 12 and 24. Interim physical and laboratory examinations were performed when indicated clinically. RESULTS: In finasteride-treated patients the total symptom score improved throughout the study, with a significant difference between the two groups at 24 months (P < or = 0.01), whereas in placebo-treated patients, there was an initial improvement in the symptom score but no change from baseline at 24 months. The maximum urinary flow rate decreased in the placebo group, but improved in the finasteride group, resulting in a between group difference of 1.8 mL/s at 24 months (P < or = 0.01). The mean change in prostate volume was +12% in the placebo group versus -19% in the finasteride treated group (P < 0.01). Finasteride was generally well tolerated throughout the 2-year study period. CONCLUSIONS: The efficacy of therapy with finasteride 5 mg in improving both symptoms and maximum urinary flow rate and reducing prostate volume has been shown to be maintained during 24 months while patients receiving placebo experienced a return to baseline or deterioration of these parameters during the study. These results demonstrate that finasteride can reverse the natural progression of BPH. PMID- 7495112 TI - Washington University experience with extracorporeal shock-wave lithotripsy of pancreatic duct calculi. AB - OBJECTIVES: To ascertain the effectiveness and safety of extracorporeal shock wave lithotripsy (ESWL) for pancreatic calculi. METHODS: Fourteen ESWL treatments were performed in 12 patients with chronic pancreatitis. RESULTS: Fragmentation was perceptible after 13 of 14 treatments. Subsequent endoscopic manipulation resulted in complete extraction, partial extraction, and failed extraction of the fragments after 7, 4, and 2 of the ESWL treatments, respectively. No complications occurred and no patient had pancreatitis following ESWL. At a median follow-up of 19 to 22 months, 4 patients have had complete relief of symptoms, 4 have had a decrease in both severity and frequency of pain, and 4 have had no improvement. CONCLUSIONS: ESWL is a safe and useful noninvasive adjunct in the treatment of patients with pancreatic duct calculi. PMID- 7495113 TI - Innocuity of agents for radiocontrast enhancement of urinary tract stones. AB - OBJECTIVES: Mineral kidney stones are frequently difficult to detect due to their radiotranslucency. We have recently developed a method that enhances the visibility of such stones by retrograde infusions of certain heavy metal salt solutions such as cesium or lanthanide gadolinium. This report describes toxicologic studies carried out on the use of those contrast agents to introduce this technique eventually into clinical trials. METHODS: Systemic absorption was assessed in dogs through infusion of radioactive contrast agent into the renal pelvis with or without ureteral obstruction. Radioactivity in urine and blood was monitored. Local toxicity was studied in animals infused with the contrast agent at intervals up to 4 weeks. RESULTS: Reabsorption studies under high intrapelvic pressures (70 cm H2O or higher), demonstrated reabsorption of cesium. However, at normal intrapelvic pressures, only a moderate reabsorption of cesium was observed. No gadolinium reabsorption was detected even at high intrapelvic pressures. Histopathologic studies showed no major urothelial lesions but only a transient inflammatory reaction that was undetectable 4 weeks following exposure to gadolinium salts. CONCLUSIONS: Gadolinium salt solutions are good positive radiocontrast agents for mineral kidney stones without having serious toxic effects or systemic reabsorption. PMID- 7495114 TI - Extracorporeal shock-wave lithotripsy of middle ureteral stones: are ureteral stents necessary? AB - OBJECTIVES: To ascertain whether insertion of a ureteral stent improves the outcome of middle ureteral (overlying the pelvic bone) stones treated with extracorporeal shock-wave lithotripsy (ESWL). METHODS: Thirty-three patients with middle ureteral stones were treated with ESWL at our institution between October 1991 and October 1994. Twenty-six patients were available for follow-up; 14 patients were treated with stent bypass, 8 were treated in situ, and 4 patients were treated after percutaneous nephrostomy (PCN). All patients were treated initially on an unmodified Dornier HM-3, and all but 4 patients were treated in the prone position on a modified Stryker frame. Follow-up consisted of a plain abdominal radiograph, intravenous urogram, occasionally a retrograde urogram, and a telephone interview. RESULTS: The overall stone-free rate for ESWL alone was 73%, and the efficiency quotient was 69. The stone-free rates after a single treatment for the stent bypass, in situ, and PCN groups were 71%, 63%, and 75%, respectively. Overall, 4% of patients required retreatment, 19% of patients required an auxiliary procedure, and 8% of the patients required hospital or emergency room admissions for renal colic. For stones 10 mm or greater (9), stone free rates after one treatment for the stent bypass, in situ, and PCN groups were 33%, 33%, and 67%, respectively; for stones less than 10 mm (17), success rates were 82%, 80%, and 100%, respectively. CONCLUSIONS: Pretreatment stinting provides no advantage over in situ ESWL for middle ureteral calculi (Fisher's exact test, P = 1.0). ESWL is a reasonable initial therapy for middle ureteral stones less than 10 mm. PMID- 7495115 TI - Dimethyl sulfoxide does not trigger urine histamine release in interstitial cystitis. AB - OBJECTIVES: Dimethyl sulfoxide (DMSO), an agent that provides symptomatic relief in patients with interstitial cystitis (IC) works via an unknown mechanism. We investigated whether DMSO acts as a chemical stimulant of mast cell degranulation. METHODS: A radioimmunoassay (RIA) specific for histamine was used to test this hypothesis. Twelve women with strictly diagnosed IC were treated with intravesical instillations of DMSO. Treatments were repeated at varying intervals, and each patient received three to six treatments. Urine histamine levels were measured before and after each intravesical instillation of DMSO. Dilutional effects of DMSO were corrected for by conversion of urine histamine concentration to urine histamine:creatinine ratio. RESULTS: The RIA was unaffected by the addition of DMSO to urine. No consistent change in the urine histamine:creatinine ratio following DMSO instillation was found. Trend analysis revealed no trend in the histamine:creatinine ratio with time. CONCLUSIONS: The relief of symptoms reported in 50% to 77% of patients treated with intravesical DMSO is not related to detectable mast cell release of histamine. Other mechanisms of action must be investigated to explain the beneficial effect of this agent. PMID- 7495116 TI - Late malignancy in bowel segments exposed to urine without fecal stream. AB - OBJECTIVES: This study was constructed so as to screen malignant transformation after uroenteric reconstructions using bowel segments exposed to urine without fecal stream for more than 10 years. METHODS: Follow-up data were available for 186 patients who underwent various uroenteric reconstructions using bowel segments exposed to urine without fecal stream for more than 10 years. There were 68 eligible patients with isolated rectosigmoid bladder, 23 with bladder augmentations (15 ileocystoplasty and 8 colocystoplasty), 57 with ileal ureter, and 38 with ileal loop conduit. Besides routine laboratory and radiologic investigations, urine for cytology was obtained from all patients. Moreover, endoscopy and random biopsy of the part of bowel exposed to urine were carried out in all patients. RESULTS: Uroenteric malignancy was diagnosed in 4 patients (2%): 2 adenocarcinoma in an isolated rectosigmoid bladder, 1 transitional cell carcinoma following augmentation colocystoplasty, and 1 squamous cell carcinoma after ileal ureter. None of the patients developed tumors in ileal loop conduits. CONCLUSIONS: Malignant changes do not only occur after ureterosigmoidostomy but are also observed after different uroenteric reconstructions not exposed to fecal stream. Hematuria, ureteral obstruction, and abnormal urine cytology are warning signs of malignancy. Routine cytology is recommended at least yearly beginning 10 years after surgery. PMID- 7495118 TI - Is prostate-specific antigen density more useful than prostate-specific antigen levels in the diagnosis of prostate cancer? AB - OBJECTIVES: To compare the performance of prostate-specific antigen (PSA) levels with the performance of PSA density (PSAD), the ratio of the serum to the size of the prostate, as predictors of the presence of prostate cancer. METHODS: We analyzed the results of digital rectal examination (DRE), transrectal ultrasonography (TRUS), serum PSA levels, and PSAD in 244 patients who had a needle biopsy of the prostate. RESULTS: Cancer was detected in 110 patients (45%). Compared with DRE, TRUS and serum PSA levels 4.0 ng/mL or higher, PSAD at a cutoff point of 0.15 ng/mL/cm3 was significantly more specific and had a higher positive predictive value than each of the other tests but was significantly less sensitive than TRUS and PSA (P < 0.05 for each). In a receiver operating characteristic analysis, PSAD was significantly more accurate than PSA (P < 0.001). In 80 patients with a normal PSA, PSAD added no additional information, and PSAD was not able to identify a subset at low risk. In 82 patients with a high PSA level (10 ng/mL or higher), 15% had a PSAD less than 0.15 and only 8% had a cancer. CONCLUSIONS: Overall, PSAD was significantly more accurate than PSA for predicting the results of needle biopsy of the prostate, but in practice PSAD proved useful in only a small subset of patients. If the serum PSA level was high but the PSAD was low, cancer was rarely detected. These patients may be suitable candidates for careful follow-up rather than early repeat biopsy. PMID- 7495119 TI - Impact of competing mortality on the cancer-related mortality in localized prostate cancer. AB - OBJECTIVES: To evaluate the impact of competing mortality and extended observation time on the cancer-related mortality in localized prostate cancer (PC). METHODS: A comparison was made between two theoretical populations of prostate cancer patients. Both populations had a slowly increasing mortality due to PC, corresponding to a 10-year cause-specific mortality of 15%. One population (A) experienced a high competing mortality reaching 50% after 10 years, corresponding to series on deferred treatment. The other population (B) experienced a low competing mortality, 10% after 10 years, corresponding to series on radical prostatectomy. The impact of these different competing mortality rates on the absolute number of patients succumbing to PC and the effect of extended follow-up to 15 years was assessed. RESULTS: The ultimate risk of death from PC after 10 years was 8% in group A and 12.3% in group B. When the observation time was extended to 15 years, group A had a 16.5% risk of cancer death and group B had a 35.3% risk. The PC mortality increased twofold between 10 and 15 years in group A (8% versus 16.5%) and threefold in group B (12.3% versus 35.3%). CONCLUSIONS: Low cause-specific mortality rates at 10 years of follow-up in series on deferred treatment comprising older patients with high competing mortality cannot be extrapolated to younger patients with a low competing mortality. Long expected survival is associated with a considerable cancer related mortality at 15 years (35%) despite favorable outcome at 10 years. PMID- 7495117 TI - Comparison of different prostate-specific antigen cutpoints for early detection of prostate cancer: results of a large screening study. AB - OBJECTIVES: This study was designed to compare the usefulness of the normal prostate-specific antigen (PSA) level and the age-referenced PSA level in a large screening study for early detection of prostate cancer. METHODS: A total of 21,078 subjects (aged 45 to 75 years) were participants in a 1-year prostate cancer screening project with PSA as the initial test. Of the volunteers, 1618 (8%) showed an elevated PSA level according to age-specific reference ranges, and using the normal PSA cutoff point (4.0 ng/mL), 1872 (9%) had elevated PSA levels between 4.0 and 6.5 ng/mL. RESULTS: Biopsies in both groups were performed if the PSA level was elevated. We evaluated the effect on biopsy rate and cancer detection. A PSA cutoff point of 2.5 ng/mL in men 45 to 49 years old and a PSA cutoff point of 3.5 ng/mL in men 50 to 59 years old with normal digital rectal examination findings resulted in an 8% increase in the number of biopsies (66 of 778) and an 8% increase in organ-confined cancer detection. An increasing cutoff of 4.5 ng/mL in men 60 to 69 years old and 6.5 ng/mL in men 70 to 75 years old resulted in 21% fewer biopsies (205 of 983) and would have missed 4% of organ confined tumors (8 of 220). CONCLUSIONS: We conclude that the use of PSA age specific reference ranges increases the detection of clinically important and organ-confined cancers in young men and decreases the number of biopsies in older men. PMID- 7495120 TI - Short-term outcomes after cryosurgical ablation of the prostate in men with recurrent prostate carcinoma following radiation therapy. AB - OBJECTIVES: Cryosurgical ablation of the prostate represents a possibly efficacious method of treating prostate carcinoma in men failing radiation therapy. In addition to eradicating the disease, cryosurgery has the potential to avoid some of the morbidity associated with other treatment modalities. Therefore, a prospective Phase II trial was conducted to determine the safety and efficacy of cryosurgery following radiation therapy in men with local recurrence. METHODS: Cryosurgical ablation of the prostate was performed in men with prostate cancer previously treated by radiation therapy. All patients had biopsy-proven recurrent disease and no evidence of metastases. Initial follow-up occurred at approximately 3 months and included repeat biopsy and serum prostate-specific antigen (PSA) determinations. Biochemical disease-free survival defined as a PSA value less than 0.3 ng/mL was assessed using the Kaplan-Meier method. RESULTS: Thus far, 23 patients have been treated with cryosurgery with follow-up ranging between 12 and 23 months. Approximately 3 months after treatment, sextant biopsies revealed no cancer in 19 of 22 (86%) of the specimens; however, 10 of 22 (45%) showed small amounts of benign glandular tissue. The PSA at 3 months postoperatively declined in 18 of 22 (82%) patients but reached female levels in only 8 men (36%). Nineteen of 22 (86%) patients followed up a minimum of 1 year after cryotherapy have a PSA level equal to or greater than 0.3 ng/mL. Complications occurred in 100% of the patients with 12 of 22 (55%) requiring at least one transurethral resection of sloughed, necrotic tissue. CONCLUSIONS: Our preliminary results suggest that the current cryosurgical technique used in men failing radiation therapy has a low probability of biochemical cure and a high complication rate. Until more data are available, cryosurgery for men with locally recurrent disease after radiation therapy should be considered strictly experimental. PMID- 7495121 TI - Loss of HLA class I expression in prostate cancer: implications for immunotherapy. AB - OBJECTIVES: There is currently no reliable predictor of the metastatic potential of apparently localized prostate cancer in an individual patient or satisfactory treatment for patients with advanced disease. One of the factors that may influence tumor progression is the cellular arm of the immune response, and central to this is the human leukocyte antigen (HLA) system, which acts to restrict T-cell recognition of potential tumor antigens. It has been reported in some cancers that down regulation of HLA class I expression by the tumor cells is associated with poor prognosis. In this report, HLA class I and II expression have been investigated in both benign and malignant prostate disease, first to define the extent of altered HLA expression and second to assess whether HLA expression may be related to disease progression. METHODS: HLA expression was assessed by immunohistochemistry utilizing a set of monoclonal antibodies that recognize both monomorphic determinants and the commoner HLA class I allelic products. RESULTS: In contrast to the normal HLA class I expression of the benign tissue, complete loss of HLA class I expression occurred in 34% of primary prostate cancers and 80% of lymph node metastases. When individual allelic expression was assessed, the minimum estimate of down regulation was 85% in the primary prostate cancers and 100% of the metastases. CONCLUSIONS: This investigation has demonstrated a higher rate of HLA class I loss than has been reported in other tumors and would suggest that the immune system may have an important role in the progression of prostate cancer, as well as having implications for the design and success of immunotherapy regimens in advanced disease. PMID- 7495122 TI - Protection of the germinal epithelium in the rat from the cytotoxic effects of chemotherapy by a luteinizing hormone-releasing hormone agonist and antiandrogen therapy. AB - OBJECTIVES: The protection of spermatogenesis during chemotherapy using an antiandrogen and a luteinizing hormone-releasing hormone (LHRH) agonist was examined in the rat. Previous studies using LHRH agonists alone have been inconclusive, as both protective and deleterious effects on the germinal epithelium have been reported. Flutamide has not previously been used in this manner but theoretically should protect the germinal epithelium, since flutamide rapidly blocks testosterone at the cellular level and also minimizes the testosterone "flare" when LHRH agonist therapy is initiated. METHODS: Mature Sprague-Dawley rats were pretreated with flutamide, sustained-release goserelin acetate (Zoladex), or a combination of flutamide and sustained-release goserelin acetate for 14 days before 4 weekly doses of procarbazine were initiated. The seminiferous tubules were evaluated histologically after a 90-day regeneration period using the stem cell assay test. RESULTS: After treatment with procarbazine alone, only 43% of the seminiferous tubules were active; however, 80% were active if protected with flutamide, 91% if protected with sustained-release goserelin acetate, and 95% if protected with both flutamide and goserelin acetate. CONCLUSION: Flutamide, sustained-release goserelin acetate, and a combination of these agents were effective in protecting the germinal epithelium of the rat during chemotherapy. A combination of flutamide and goserelin acetate provided the best protection. This study demonstrates for the first time the protective effect of flutamide and flutamide with goserelin acetate on the germinal epithelium during chemotherapy. PMID- 7495123 TI - Electrovasogram: a canine study of the electromechanical activity of the vas deferens. AB - OBJECTIVES: To study the electric activity of the vas deferens (VD), or electrovasogram (EVG), in 14 mongrel dogs. METHODS: The VD was exposed in the spermatic cord and three electrodes were applied to it. Synchronous transcutaneous EVG was also recorded by another three electrodes. The electric activity and intravasal pressure (IVP) were determined at rest, during ejaculation, and after vasal myotomy. RESULTS: Slow waves or pacesetter potentials (PPs) were recorded regularly from all electrodes applied to the VD whether directly or transcutaneously. They had identical frequency and regular rhythm from all the electrodes and were consistent in the individual dog on all test days. Action potentials (APs) followed PPs randomly and were associated with an IVP increase, indicating a state of contractile activity. Just before ejaculation, the PPs and APs increased significantly (P < 0.05) and were associated with IVP rise (P < 0.01). During ejaculation, there was rhythmic increase of the PPs, APs, and IVP (P < 0.01 for frequency and amplitude, P < 0.05 for velocity of PPs and P < 0.001 for IVP) followed by a silent period and return to the resting activity (P > 0.05). After vasal myotomy, PPs and APs were recorded proximal but not distal to the cut. This suggests that the PPs and APs spread caudally from the proximal part of the VD that is believed to be the site of a pacemaker that triggers the PPs. The PPs seem to pace the vasal contractile activity in terms of direction and frequency. CONCLUSIONS: A normal EVG pattern was characterized for the VD, which may be deranged in the various vasal pathologic conditions. EVG may thus be proposed to be included as an investigative tool in the diagnosis of vasal disorders. PMID- 7495125 TI - Ventral penile curvature following radical pelvic surgery: a variant of urethral manipulation syndrome. AB - OBJECTIVES: Urethral manipulation syndrome (UMS) describes ventral penile curvature and urethral stricture disease that develop following repeated episodes of urethral manipulation. We describe a variant of this syndrome, in which the presence of an indwelling catheter following radical pelvic surgery led to a marked penile curvature without clinically apparent urethral stricture disease. METHODS: We identified 4 patients in whom ventral penile curvature developed following radical pelvic surgery. Three patients underwent radical retropubic prostatectomy and the fourth patient underwent radical cystoprostatectomy with creation of a neobladder to the urethra. All were treated with 3 weeks of catheter drainage postoperatively. Each patient reported straight erections prior to surgery. RESULTS: These patients came to prosthesis surgery between 7 months and 3 years after their pelvic procedure. Each patient was noted to have ventral penile curvature when artificial erection was induced. Curvature was secondary to scarring between the anterior corpus spongiosum and the overlying ventral tunica albuginea in the mid- to proximal penile shaft. It was necessary to mobilize the urethra off the corpora and to incise the scarred tunica to obtain a satisfactory result at the time of prosthesis placement. In 3 cases, GoreTex was needed to bridge the corporeal defect. CONCLUSIONS: These cases represent a variant of UMS in which catheter drainage leads to scarring of the ventral tunica albuginea, resulting in ventral penile curvature. In view of the increasing number of radical pelvic procedures being performed, this potential complication must be recognized, as aggressive corporeal reconstruction with urethral mobilization is needed if subsequent prosthesis surgery is undertaken. PMID- 7495124 TI - A brief male sexual function inventory for urology. AB - OBJECTIVES: To develop a brief questionnaire to measure male sexual function. METHODS: An initial set of questions was refined and reduced through cognitive testing and two serial validation studies. In each study, men were recruited from a sexual dysfunction clinic and a general medicine practice to complete the instrument. Test-retest reliabilities, internal consistencies, and construct validities were examined. RESULTS: The final instrument covers sexual drive (two items), erection (three items), ejaculation (two items), perceptions of problems in each area (three items), and overall satisfaction (one item). Psychometric performance was generally very satisfactory, although self-assessments of ejaculate volume are problematic. Translations have been developed and pilot tested in a number of languages. CONCLUSIONS: The Brief Sexual Function Inventory may be useful for measuring male sexual function in practice and research. PMID- 7495126 TI - Penile cancer--is lymphadenectomy necessary in all cases? AB - OBJECTIVES: This retrospective study examines the need for lymphadenectomy in all cases of penile cancer. METHODS: Thirty-six patients were followed up regularly by the same person between 1986 and 1992. RESULTS: Nine (25%) patients presented with positive groin lymph nodes and underwent immediate ilioinguinal node dissection. Of the 27 (75%) patients who presented with negative groin lymph nodes, 10 had a delayed lymph node dissection. Four of these patients are alive at 3.7 years. Three died within 1 year of surgery and 3 are lost to follow-up. Seventeen patients (47%) did not require lymphadenectomy and 14 (82%) of these patients are alive at 3.8 years, the other 3 being lost to follow-up. CONCLUSIONS: A careful, closely monitored, follow-up protocol can eliminate the need for lymphadenectomy in select patients with penile cancer. PMID- 7495127 TI - The laparoscope as a teaching aid in open pelvic surgery. AB - Laparoscopic imaging equipment is useful in open surgical procedures, especially in the deep male pelvis. The magnification and brilliant illumination provided by these unobtrusive instruments increase visibility, facilitating both dissection and reconstruction. The application of this technique for surgical education is discussed. PMID- 7495128 TI - Laparoscopic orchidopexy. PMID- 7495129 TI - Renal arteriovenous malformations masquerading as renal cell carcinoma. AB - Renal arteriovenous malformations may have varied clinical and radiographic appearances. Often, it remains difficult to distinguish these lesions from renal cell carcinomas to tailor the most appropriate diagnostic evaluation and therapy. We have encountered 6 patients with renal arteriovenous malformations that masqueraded as renal cell carcinomas and describe their clinical presentation and management. PMID- 7495130 TI - Renal cell carcinoma metastatic to the gallbladder or prostate: two case reports. AB - Although metastatic renal cell carcinoma (RCC) is not uncommon, dissemination to the gallbladder or prostate is very rare. We present 1 case of RCC with synchronous metastasis to the gallbladder and 1 case of RCC with metachronous metastasis to the prostate. These cases illustrate the propensity of RCC for unpredictable presentation and unusual sites of metastases. The first case also illustrates the benefit of adjuvant nephrectomy with excision of a solitary metastasis. Patterns of RCC metastases and flow cytometric analysis of RCC are discussed. PMID- 7495132 TI - Intravesical thiotepa-induced eosinophilic cystitis. AB - Eosinophilic cystitis is a rare form of an allergic cystitis. Factors such as food allergens, parasites, and drugs have been implicated in the genesis of eosinophilic cystitis. Associated risk factors include bronchial asthma, atopic diseases, and environmental allergens. Intravesical mitomycin is a reported causative agent. We report the first case of eosinophilic cystitis that developed after intravesical instillation of thiotepa for treatment of superficial bladder cancer. PMID- 7495131 TI - Colonic duplication: an unusual cause of enterovesical fistula. AB - Intestinal duplications are rare, occurring in 1 in 4000 births. Even rarer are tubular hindgut duplications, with less than 75 cases reported in the literature to date. We report an uncommon case of an enterovesical fistula caused by a duplication of the sigmoid colon. This is the first reported case of a tubular colonic duplication presenting as a colovesical fistula in an adult. We discuss the presentation and shortcomings in the diagnosis and treatment of enterovesical fistulas and colonic duplications. PMID- 7495133 TI - Eosinophilic granulomatous cystitis in children. AB - We report 3 cases of eosinophilic granulomatous cystitis in children. All presented with hematuria, urinary urgency, and dysuria. Treatment with oral corticosteroids led to rapid and complete recovery in 2 of the 3 cases. In the third case recovery was prolonged without the use of steroids. Because of the presumed immunologic nature of this disease, previous authors have recommended the use of corticosteroids in its management. A review of the literature suggests that the disease is probably of immunologic nature and that corticosteroids shorten symptomatic recovery from this disease. PMID- 7495135 TI - Detached ciliary tufts in the epididymis: a lesson in applied anatomy. AB - We report a case of detached ciliary tufts (DCTs) discovered in the aspirate from an epididymal cyst in a 30-year-old man undergoing epididymal exploration for obstructive azoospermia. The specimen was initially misdiagnosed as a parasite due to its appearance and spontaneous motion on fresh smear. Since standard urology and histology texts identify ciliated cells only in the efferent duct of the male genital tract, the source of DCTs in this patient was mysterious. However, on further review, histologic studies have demonstrated that the first portion of the epididymis represents an extension of the efferent ducts and is lined with cilia. The fact that the caput epididymis is composed of branched efferent ducts has significant implications for our understanding of congenital absence of the vas, epididymal sperm aspiration, spermatoceles, and epididymal physiology. PMID- 7495134 TI - True hermaphroditism and normal male genitalia: an unusual presentation. AB - True hermaphroditism, the rarest form of intersex, is usually diagnosed during the newborn period in the course of evaluating ambiguous genitalia. As an exception, we present an unusual case of a 15-year-old boy with phenotypically normal male genitalia and bilaterally descended ovotestes, who was seen for evaluation of intermittent scrotal swelling and pain. Although rare, true hermaphroditism should be included in the differential diagnosis of apparently normal males presenting with similar complaints. PMID- 7495136 TI - Antegrade ejaculatory duct recanalization and dilation. AB - Ejaculatory duct obstruction is a rare but correctable cause of male infertility. Standard therapy is transurethral resection of the ejaculatory ducts, which is frequently complicated by seminal vesicle urinary reflux and is contraindicated when the obstruction is located outside the prostate gland. Herein, we report a minimally invasive technique that successfully dilated the ejaculatory duct without complications in a patient with unilateral, complete ejaculatory duct obstruction located outside the prostate. PMID- 7495137 TI - Initial experience with retroperitoneoscopic nephroureterectomy: use of a double balloon technique. AB - OBJECTIVES: To report the initial experience with retroperitoneoscopic nephroureterectomy for symptomatic, end-stage vesicoureteral reflux. METHODS: Two patients underwent a nephroureterectomy by a four-port retroperitoneal laparoscopic approach. In 1 patient, a double-balloon technique was used to dissect the pelvic extraperitoneal space and gain access to the juxtavesical ureter. In the second patient, the distal ureter was mobilized with routine laparoscopic dissection techniques. RESULTS: Operative time was 6 and 5.5 hours, respectively, and mean hospital stay was 4 days. Mean patient follow-up is 17.5 months. CONCLUSIONS: The technique of retroperitoneoscopic nephroureterectomy is in evolution; until now, a major concern has been the inadequate access to the distal ureter through a completely retroperitoneoscopic approach. Described herein is a double-balloon technique that significantly facilitates dissection of the juxtavesical ureter during a retroperitoneal laparoscopic nephroureterectomy. PMID- 7495138 TI - Percutaneous electrovaporization of upper tract transitional cell carcinoma in patients with functionally solitary kidneys. AB - OBJECTIVES: We present the first reported experience using percutaneous electrovaporization with both a rollerball and the VaporTrode electrode of large upper tract transitional cell carcinoma (TCC) in 2 individuals with functionally solitary kidneys. METHODS: Under general anesthesia and in the prone position, 2 patients underwent electrovaporization of upper tract TCC through upper pole percutaneous access. Using either a rollerball (Case 1) or the VaporTrode (Case 2) electrode at 200 W pure cutting current, the tumors were completely vaporized. RESULTS: Average operating room time was 145 minutes. Hemostasis was excellent. No significant perioperative complications were noted. A nephrostogram at the end of the procedure revealed no extravasation in either case. There was no appreciable change in serum creatinine. Both patients received postoperative mitomycin (40 mg in 1000 cc normal saline over 20 hours) through the nephrostomy tube. One patient (Case 2) is completely free of disease via antegrade nephroscopy and biopsy at 2 weeks' follow-up. The other patient (Case 1) had two remaining focal areas of tumor via antegrade nephroscopy 3 weeks postoperatively; both lesions were treated with electrovaporization via a flexible electrode through the flexible cystoscope and the VaporTrode, respectively. CONCLUSIONS: Percutaneous electrovaporization of upper tract TCC appears to provide a simple, rapid, and effective endosurgical treatment in properly selected patients. Long term follow-up will be needed to assess this new treatment modality definitively. PMID- 7495139 TI - Breastfeeding for optimal mental development. The alpha and the omega in human milk. PMID- 7495140 TI - Breastfeeding in Korea. PMID- 7495142 TI - Breastfeeding trends in Cuba. PMID- 7495141 TI - Breastfeeding in China. PMID- 7495143 TI - Breastfeeding patterns in the Arabian Gulf countries. PMID- 7495144 TI - Breastfeeding kinetics. A problem-solving approach to breastfeeding difficulties. PMID- 7495145 TI - Human milk and premature infants. PMID- 7495146 TI - Breastfeeding in Australia. PMID- 7495147 TI - [The organized brain (an essay on the principles of the design and functional organization of the brain). I]. AB - The study of correlating the principles of the cerebral functional organization with the main mechanisms of the behavioral act is based on the necessity of joining analytical and synthetic approaches. The analysis of interrelationships between micro- and macrolevels of brain organization as well as study of the role of their relations in architecture of concrete behavioral reaction is one of the perspective ways to this problem. Two basic forms of construction and activity rigid, genetically determinate and flexibly individual--are constantly interacted in the living brain as well as in whole organism. This essay is dedicated to the development of original author's conception concerning the some structure functional characteristics of different brain systems and main principles of organization and integration of cerebral functions. PMID- 7495148 TI - [The amygdaloid body of the brain: some topics for discussion and little-studied problems]. AB - Features of functional morphology of the amygdaloid brain complex (ABC) are analyzed in a new light presenting the notion of the ABC as a nucleo paleocortical structure. The analysis of the structural organization of ABC could imply the presence of phylogenetically differentiated parts: the ancient, the old and the new amygdala. The presentation carries original data on the morphometric and histophysiological studies under various experimental alterations of the endocrine system. A concept is developed of the existence of a rostrocaudal gradient in the expression of sex-dependent structural and functional features of the amygdaloid complex as a neuroendocrine brain centre. Provided is technical recommendation for use in biological studies of amygdaloid brain complex mathematical models based on the theory of image identification. An original technique has also been recommended for selective dissection of the ABC with its eventual removal from nonfixed brain, which allows studies to be made of the ABC neurophysiological and neurochemical properties on surviving slices. Evidence is adduced indicating involvement of the ABC in the process of the brain sex differentiation. PMID- 7495149 TI - [The regulation of food consumption (the ontogenetic aspects)]. AB - The present-day concepts of organizing the physiologic system of food consumption control at different stages of ontogenesis of the premature born mammals are discussed with an element of criticism. The "alimentary center" notion as a multilevel system of the perceptive neurons lost its original significance and needs improvement. An idea of hypothalamic alimentary centers also needs revision to match the recent detection of many brain structures which are associated with the development of hunger and repletion states. There is no strong evidence of a direct involvement of central reception of metabolic products in an immediate control of the alimentary behavior. The hunger and repletion theories each only in part explain the physiologic basis of these states. The alimentary behavior is an integrated process not coming to a consummatory act. It relates to a complex interaction of afferent and efferent systems which are different at various stages of ontogenesis. In an early period of life the regulation of alimentary behavior depends mainly on the sensory signals and only gradually it incorporates the metabolic canal of signalling. A delay in the repletion mechanisms provides the optimal conditions for the growth and development of organism. The formation of alimentary stereotypes on the basis of specific and nonspecific factors is described. Regulation of different aspects of alimentary behavior depends finally on their relation to the state of brain systems of reward and punishment. PMID- 7495150 TI - [The participation of enteral mechanisms in the sympathetic-adrenal inhibition of gastric secretion]. PMID- 7495151 TI - [The calcium channels of cell membranes]. AB - On the basis of the literature findings, the modern ideas about the structure and function of calcium canals of plasmatic and intracellular membranes of the living cells are generalized and systematized. The major electrophysiologic and chemical properties of the main groups and types of potential-controlled and receptor controlled calcium canals are reviewed. The article gives the classification and nomenclature of the calcium canals as well as the characteristics of different types of calcium antagonists and the cases of their clinical application. PMID- 7495152 TI - [Epidemiology of rheumatologic health care in Germany]. AB - Within the governmental funding program of rheumatological centers a uniform patient registration is performed. It provides information of the proportion of patients with inflammatory rheumatic diseases who are seen in specialized care, how early they are seen and which forms of treatment are provided. In 1993 in the 20 participating centers almost 26,000 patients have been recorded. The centers only reach a small proportion of the true prevalence but they see their patients early in the disease: 60% of the patients have been seen within the first year. Deficits exist in the areas of ergotherapy, psychological support and patient education. One reason for the small proportion of people with the diseases who are seen in specialized care is the lack of rheumatologists in outpatient care. More rheumatologists in private practices as well as the participation of clinical rheumatologists in outpatient care including reasonable forms of payment are needed. PMID- 7495154 TI - [The established rheumatologist in the tension field between health care deficits and cost control in public health]. AB - The outpatient treatment of rheumatoid patients in Germany depends on certain conditions. Rheumatologists in Germany need to cooperate much more with hospitals specialized in this field than other subspecialties of Internal Medicine. Because of historical reasons many other physicians such as orthopedics, internists, general practitioners participate in this field in contrast to our neighbours in the western hemisphere, where rheumatology is solely covered by rheumatologists. It is therefore conceivable to calculate a rheumatologists/population relation of appr. 1:200,000 and not less than this figure. Starting a business as rheumatologist needs certain conditions, which are specified. One major challenge will be the new law directing out-patient treatment in Germany (Gesundheitskontrollgesetz). It forces the doctor to relate his work to a more financial background with various restrictions and regulations. In this context our goal(s) must be the institution of a specified position in the "Gebuhrenordnung", to pay for our intense and time consuming efforts to treat our patients. PMID- 7495153 TI - [Basic principles of community, continuous and cooperative management of patients with chronic rheumatic diseases in Germany]. AB - The lecture presents foundations and results of the work of a commission of the German Rheumatological Association. Based on epidemiological data on the occurrence of selected rheumatic disorders, reference figures for both, in- and outpatient services are developed. There is need of at least one full-time rheumatologist for every 150,000 residents. The commission identifies various actual deficits in the care of rheumatic patients. The most important seems to be the insufficient number and unequal spatial distribution of practicing rheumatologists. Other deficits concern the imperfect integration of preventive, curative and rehabilitative services, of in- and outpatient care and of the work of primary and secondary physicians with various allied health professionals at the community level. PMID- 7495155 TI - [Responsibilities and goals of a rehabilitation clinical in the present and future]. AB - Rehabilitation its an important part of the management of patients with chronic rheumatic diseases and it aims to improve or supervene physical or psycho-social disability and handicap. With regard to the fluctuating course of chronic inflammatory rheumatic diseases, a universal concept of rehabilitation always includes the curative aspect of treatment. Based on the legal and organizational premises for rehabilitation, the problems of access, usage, management and of continuous treatment are critically reviewed. The important tasks in rehabilitation of patients with rheumatic diseases by rehabilitation clinics are education and informing of patients about their disease, including the outcome, possible treatments, self management and coping strategies as well as functional therapy of disorders of the locomotor system. In addition, the adaptation of medical and physical treatment, advice and help with regard to work and occupation, and planning of continuing rehabilitative interventions are important activities of rehabilitation clinics. In the future, an improved integration of rehabilitation within concepts of medical and social support, progress in quality of structuring and processing activities in rehabilitation clinics, and the expansion of ambulant rehabilitation to facilitate continuous support has to promoted. PMID- 7495156 TI - [Patient education--a contribution to improvement of long-term management of patients with rheumatism]. AB - Patient education has been recognized as an important tool in the therapy and the rehabilitation of patients with chronic rheumatic diseases. An increasing body of literature refers to favorable effects of patient education. Patient education improves coping with disease, reduces pain and depression and decreases arthritic "helplessness". Therefore, patient education should be a part of the comprehensive care for the rheumatic patient. The patient education group (Arbeitskreis Patientenschulung) of the German Society of Rheumatology (Deutsche Gesellschaft fur Rheumatologie) has been initiated to develop a general concept of patient education in rheumatology and to elaborate various patient education programs. The patient education program for rheumatoid arthritis is described as a model for further patient education programs. It consists of six group sessions (modules). The program is conducted by an interdisciplinary team. A group leader is in charge of the program. Train-the-trainer courses should improve the quality of the program. Currently, the patient education group develops education programs for patients with systemic lupus erythematodes and related collagen diseases, spondyarthritis patients, patients with fibromyalgia and rheumatic children and their parents. PMID- 7495157 TI - [Rheumatologic patient management and research in Berlin at the intersection between East and West]. AB - After the fall of the Berlin Wall in November 1989, there was an immediate need for collaboration between rheumatologists from East- and West-Berlin from the very beginning, which could be taken as an example for the rest of Germany. This was achieved through different activities in patient care and research. The "Regionales Rheumazentrum Berlin" which was started at about that time played a special role in making this enterprise a success. PMID- 7495158 TI - [A cooperative model of the German Ministry of Health funded rheumatic disease centers]. AB - The German Ministry of Health has granted since 1992 the institution of 21 multipurpose arthritis centers in 21 regions of Germany to improve the integrated care for patients with chronic rheumatic diseases. To coordinate and combine their efforts the supported rheumatic centers have joint together into a model cooperation. The basic founding for all centers aim to advance the interdisciplinary care network inside the centers, to elaborate appropriate measures of audit, to test new modalities for better care and to advance post graduate and continuing medical education. In addition, continuing funding supports individual projects (n = 5) and since 1994 in 5 of the centers the German Ministry of Research and Technology granted the institution of integrated rheumatological research projects. The development and effects of the multipurpose arthritis center programme is evaluated by the Scientific Institute of the German Medical Association. The successful structural development of the centers and the first results of the joint efforts of the model cooperation (recommendation for surveillance of treatment with long acting antirheumatic drugs, basic documentation, data set for documentation of rheumatological out patients) shows the promising outcome of the programme. PMID- 7495159 TI - ["State of the art" in epidemiology of arthroses. An overview of important studies of etiology and sequelae]. AB - The term "Osteoarthrosis" encompasses a group of joint disorders characterized by pain and restriction of mobility. Currently, considerable confusion prevails with respect to the definition of the disease entity and the classification criteria to be utilized in epidemiologic research. Despite these methodological difficulties, it is discernible from the international rheumatology journals that during the past decade this emerging field of research has become firmly established in several European countries and in North America. Researchers have focused their attention on four major facets of the disease: 1. etiological or risk factors with respect to the initiation and clinical course of disease, 2. the impact of illness on the individual and his family, 3. the statistical estimation of incidence and prevalence rates in the total population or in selected at-risk groups of individuals, 4. health care utilization patterns and their costs and benefits. This survey article aims at providing a critical overview of this emerging field of research, as well as at suggesting to the reader future lines of development useful for public health and health promotion researchers in Germany. PMID- 7495160 TI - Cartilage destruction in septic arthritis--electron microscopy and historical considerations. AB - Light and electron microscopic investigations of joint cartilage in 13 cases of septic arthritis revealed sites of cartilaginous matrix infiltrated with polymorphonuclear granulocytes. This finding confirms rather unknown morphological works done by investigators on this diseases in the late 19th and beginning of the 20th century. Around neutrophils ultrastructural signs of direct chondrolytic activity were observed. In three cases bacteria were discovered inside the cartilaginous matrix. It is assumed that the invasion of polymorphonuclear granulocytes into the cartilaginous matrix is an important process in septic arthritis leading to cartilage destruction. PMID- 7495161 TI - Recurrent bilateral insufficiency fracture of the talus. A diagnostic pitfall for ankle pain. AB - We report on a 75-year-old woman who presented recurrent bilateral ankle pain and swelling related to insufficiency fractures. The diagnosis of this rare condition is difficult since the initial X-rays are usually normal. We may expect an increased frequency of insufficiency fractures of unusual bones with aging of the population. PMID- 7495163 TI - Abomasal displacement in the bovine--a review on character, occurrence, aetiology and pathogenesis. AB - Literature on the subject of character, history, occurrence, causes and pathogenesis of abomasal displacement in the bovine is listed and summarized. The chapter on occurrence is subdivided according to heritibility, sex, age, season, husbandry, milk yield and concomitant diseases. In the chapter on causes special consideration is given to feeding. The chapter on pathogenesis is subdivided according to mechanical influences, disturbances of the abomasal motor activity, collection of gas and dilatation of the abomasum and artificial displacement of the abomasum. Displacement of the abomasum is seen predominantly in the spring in herds with above-average milk yield in dairy cows between the age of 4 and 7 years which have calved within the last month and have ketosis. The etiology and pathogenesis of abomasal displacement are unclear. PMID- 7495162 TI - [Current aspects of SAPHO syndrome]. PMID- 7495164 TI - [Description and determination of the diameter of arteries and veins in the hindlimb of cattle using B-mode ultrasonography]. AB - The large arteries and veins of both metatarsal and digital regions of 14 healthy, non-pregnant cows with a mean age of 6.1 +/- 1.7 years were examined sonographically (real-time B-mode system with a 7.5 MHz linear-transducer). The ultrasonographic imaging of the larger vessels was investigated, diameters were measured, especially of the digital veins, in which intravenous regional anaesthesia and antibiosis is usually applied. Three measurements per cow on each distal hindlimb were noted over a period of 3 weeks using defined anatomical locations. The mean diameter of the vessels were as follows: Arteria (A.) dorsalis pedis 4.1 +/- 0.4 mm, Vena (V.) dorsalis pedis 5.2 +/- 0.5 mm, A. metatarsea dorsalis III 3.3 +/- 0.4 mm, A. digitalis dorsalis communis III 3.9 +/ 0.4 mm, V. digitalis dorsalis communis III 2.7 +/- 0.4 mm, V. digitalis plantaris communis IV 2.1 +/- 0.4 mm, V. digitalis plantaris communis II 2.1 +/- 0.3 mm, V. metatarsea plantaris lateralis 4.5 +/- 0.8 mm, V. metatarsea plantaris medialis 4.4 +/- 0.6 mm. The Ramus cranialis of the V. saphena lateralis, the A. and V. digitalis plantaris communis III and the A. digitalis plantaris communis II and IV could only be imaged in a few cases. Except for V. digitalis dorsalis communis III, statistical analysis of the mean diameters of the examined vessels of the right and the left distal hindlimb in 14 cows revealed no significant differences (P < 0.05). B-mode ultrasonography proved a suitable method for imaging and examination of the vessels larger than 1 mm in the distal bovine hindlimb and can be recommended as the non-invasive method of choice for the examination of blood vessels in cattle. PMID- 7495165 TI - A standard water provocative test for dogs anaesthetized with acepromazine ketamine. AB - Administration of 60 ml/kg of water to anaesthetized dogs increased intraocular pressure; this reached its maximum at 15 min after ingestion, except in young dogs, whose maximum was attained at 10 min. Pressure fell back close to original levels by 60 min. Pressure increases in females and young dogs were greater than those of males and adults, respectively. PMID- 7495166 TI - Influence of endometrial cysts on conception rate of mares. AB - A total of 259 normally fertile mares were examined gynecologically by means of rectal palpation and ultrasonography in order to record the presence of uterine cysts and pregnancy. The incidence of endometrial cysts was 22.4%. Of the 95 cysts observed during the trial, 87.4% were located in the middle and posterior segments of both uterine horns. The size of all endometrial cysts ranged between 3 and 48 mm. When all mares were assigned to three age groups, A < 7 years (n = 116), B 7-14 years (n = 117) and C > 14 years (n = 26), a significant (P < 0.01) increase in the number of endometrial cysts was observed with advancing age (4.3%, 29.1% and 73.1%, respectively). The pregnancy rates at Days 14 and 40 were significantly (P < 0.01) lower in mares with cysts (77.6% and 71.4%) compared to mares without cysts (91.5% and 88.0%). This suggests that the presence of uterine cysts plays an important role in the reduction of fertility of Thoroughbred mares. PMID- 7495167 TI - Ovulation as determined by transrectal ultrasonography in multiparous sows: relationships with oestrous symptoms and hormonal profiles. AB - Twenty-four cross-bred multiparous sows in their second and fourth parities were studied during two consecutive oestruses after weaning. Careful oestrous control was done on the animals and ovulation was monitored by transrectal ultrasonography. Blood for analysis of LH and E2 contents was collected through an indwelling catheter. The sows were mated/inseminated once approximately 18 h before expected ovulation in their second oestrus. The interval between weaning and ovulation varied from 86 h-204 h (153 +/- 26). Onset of standing oestrus to ovulation was found to be longer in sows with long oestruses and ovulation occurred within a mean of 68% of the oestrous period. Duration of the first pro oestrus was significantly shorter than the second pro-oestrus (40 h and 71 h, respectively). There was no significant difference within sows for the intervals from onset of oestrus to ovulation during the first and second oestrus after weaning (37 h and 39 h, respectively). The diameter of the largest follicles at onset of oestrus and at ovulation had very small variation between animals. The mean intervals from E2 peak to onset of oestrus and to ovulation were 1 h (range 10-(+)22 h) and 44 h (range 34-54 h) respectively, and from LH peak to onset of oestrus and to ovulation 12 h range -10-(+)32 h) and 35 h (range 27-48 h), respectively. The present study indicates that repeated ultrasonographic examination of ovaries during one oestrus can be used to predict the time of ovulation during the subsequent oestrus.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7495168 TI - Modulation of endotoxin-induced andrological alterations by flunixin meglumine in the boar. AB - Flunixin meglumine (FM), an inhibitor of the cyclo-oxygenase pathway of the prostaglandin synthesis, was used to evaluate the andrological changes following an endotoxin (ET) administration in the boar. FM was injected as a single dose 10 min prior to an ET injection. Blood plasma was analysed for the contents of 15 ketodihydroprostaglandin (PG) F2 alpha LH, testosterone and cortisol. Twelve h after the ET administration, the boars were castrated and the testes examined by light and electron microscopy. The results were compared between controls, ET-, FM- and FM+ET-treated boars. An increase in 15-ketodihydro-PGF2 alpha levels was seen following ET administration. Pretreatment with FM decreased the levels of 15 ketodihydro-PGF2 alpha during 2 h. Following the ET administration a voluminous increase in testosterone levels as well as changed LH levels were seen. Similar alterations in hormonal levels were observed also after pretreatment with FM, however, delayed about 2 h, and during this time a marked decrease in testosterone levels was seen. In the testes an infiltration of polymorphonuclear neutrophils (PMN) in the interstitium and histological changes among the Leydig cells were seen after ET injection. Pretreatment with FM reduced the number of invading PMN and the Leydig cells appeared less affected. The results show that FM modulated the negative effects of ET on the testicular function in the boar, indicating that the initiation of the arachidonic acid cascade was of importance for the induction of the alterations following an ET injection. PMID- 7495169 TI - Investigations on preileal digestion of starch from grain, potato and manioc in horses. AB - In this study preileal starch digestibility of starchy feeds (oats, corn, barley, potatoes, manioc) was determined in seven jejunofistulated horses. The grains were fed whole (oats, corn), rolled (oats, barley), crushed, ground and expanded (corn); the potatoes were fresh, the manioc rolled. Ground corn was also fed in combination with amylase. The feeds were fed partly isolated or in combination with alfalfa meal or hay (Table 1). At least four horses with a cannula in the terminal jejunum were used for each diet. Two meals per day were offered at 12 h intervals. The starch intake was mostly about 2 g/kg bw/meal, except one period with oats (3.9 g starch/kg bw) and with expanded corn (1.4 g/kg bw). Jejunal chyme was postprandially collected 11 times (from 1st to the 11th h after the morning meal for 15 min). Starch was determined polarimetrically. The preileal digestibility of starch was calculated by the marker method (chronic oxide 0.25% DM) and by estimating the total jejunoileal chyme flow during 12 h postprandially extrapolating the sample volume from the 15 min sampling periods. The results of both methods agreed quite well. Preileal digestibility of oat starch (80-90%) was (independent of doses or preparation or of the combination with hay, Table 4) significantly higher than that of whole or crushed corn (30%) or barley (26%). Grinding of corn significantly increased preileal digestibility to 51%, expanding to 90%. The addition of amylase improved digestion of ground corn by 10% (absolute). The preileal digestibility of potato or manioc was less than 10%. Individual factors in the horse (chewing intensity, amylase activity) had also considerable influence on preileal starch digestibility. PMID- 7495170 TI - Age-related morphometrical changes of arteries of uterine wall in mares. AB - To investigate the morphological changes of uterine wall arteries associated with the progression of age in Thoroughbred broodmares, uterine wall arteries from 13 mares, aged 2 to 31 years, were studied by routine histomorphometry. The ratio of intimal thickness to external diameter increased due to hyperplasia of elastic fibers and was significantly correlated with age (r = 0.95, P < 0.001). The ratio of medial thickness to external diameter decreased and was also significantly correlated with age (r = -0.84, P < 0.001). Using electron microscopy, it was determined that the cross-sectional area of the tunica media occupied by smooth muscle cells gradually decreased with increasing age. These results demonstrate that in arteries of the uterine wall in mares, hyperplasia of intimal elastic fiber and thinning of the medial smooth muscular layer is associated with aging, suggesting that aging may influence endometrial pathology. PMID- 7495171 TI - Prochymosin polymorphism in calves of black-and-white cattle and their crosses with Simental bulls. AB - Polymorphism of prochymosin was observed in individual calf abomasa, using agarose gel electrophoresis followed by detection of proteolytic activity. Abomasum samples were randomly collected during slaughtering from 239 and 146 calves (3-5 weeks old) of Black-and-White cattle and their crosses with Simental bulls, respectively. Four distinct prochymosins were found and, according to their decreasing electrophoretic mobility in alkaline agarose gel, termed as prochymosin A, D, B and C which occurred singly and in pairs (then with equal proteolytic activities of both components). Prochymosin A, B and C (designation according to FOLTMANN, 1966) activated at pH 4.7 was transformed into electrophoretically distinct chymosin. When prochymosin D was activated at this pH, chymosin D showed similar mobility as chymosin B both at alkaline and acidic pHs. Prochymosin variants occurred at genetical equilibrium in nine and ten phenotypes in the first and second genetic group. The distribution of phenotypes in the two groups differed significantly (P < 0.05). The gene frequencies of prochymosin A, D, B and C were 0.35, 0.11, 0.52 and 0.02 in Black-and-White calves, and 0.39, 0.08, 0.47 and 0.06 in crosses, respectively. These prochymosins were controlled by four pairs of codominant alleles. A possible correlation of the results obtained by FOLTMANN (1966) with ours and those of ASATO and RAND (1972, 1977) was discussed. PMID- 7495172 TI - Long-term treatment of diseased, FIV-seropositive field cats with azidothymidine (AZT). AB - The long-term effectiveness of Azidothymidine (AZT, Zidovudine) was evaluated in FIV-seropositive cats with clinical symptoms (n = 9; Group 1) compared with conventional symptomatical therapy (n = 5; Group 2). The oral administration of Azidothymidine at a dosage of 5 mg/kg BW yielded a mean peak plasma concentration of 4.59 microM one hour after application (median: 3.74 microM). Elimination half time was 1.5 h. The permanent oral application of Azidothymidine at a dosage of 5 mg/kg BW TID led to a total recovery from clinical symptoms in six of nine FIV seropositive cats (Group 1) 4-6 weeks after the onset of therapy. One cat clinically improved with only sporadical recurrence of disease; therapy with Azidothymidine was not effective in two cats. All the FIV-seropositive cats treated symptomatically responded well to antibiotics and immunomodulators within 10-14 days (n = 5; Group 2). Recurrence of clinical symptoms was noticed in three of five patients within 2 years after therapy and one cat died. During the treatment with Azidothymidine hyperproteinemia and abnormal albumin-to-globulin ratio became normal within 6 months in four FIV positive cats (Group 1), whereas hyperproteinemia did not change in Group 2. The following adverse effects were noticed in Group 1: a transient decrease of red blood cell count (RBC), packed cell volume (PCV) and haemoglobin until the 4th week of application of Azidothymidine (5/8), but the haemogram was within the normal range after 3 months. In one cat with hyperglycemia the anaemia remained until the administration of Azidothymidine was terminated. Heinz (Schmauch) bodies in the erythrocytes appeared in two FIV-positive cats 2 weeks after the onset of the therapy. PMID- 7495173 TI - Binding properties of goat intestinal vitamin D receptors as affected by dietary calcium and/or phosphorus depletion. AB - The binding capacity (Bmax) and the affinity (Kd) of the intestinal vitamin D receptor (VDR) have been studied using mucosa preparations from the duodenum, jejunum and proximal colon of male growing goats which had been kept in a two factorial (2 x 2) trial on Ca and/or P deficient diets for 9 weeks. This treatment resulted in significant changes of different parameters of Ca and P homeostasis. Irrespective from the level of Ca intake, P depletion caused significant hypophosphatemia with corresponding hypercalcemia. In both Ca depleted groups the calcitriol concentrations in plasma significantly increased by more than 100% in comparison with normal Ca supply. No changes were recorded for plasma calcitriol concentrations in response to P depletion with an adequate Ca supply. Plasma PTH levels were only increased significantly in Ca depletion with adequate P supply. Irrespective of different feeding regimens, the highest Bmax values were found in the jejunum. In all intestinal segments tested, the Bmax values were significantly decreased by P depletion as compared with an adequate P supply. No effects on the Bmax of VDR were observed in response to changes of Ca supply. The Kd values of the VDR were neither affected by different intestinal localizations nor by Ca and/or P depletion. From the present results, it has to be concluded that the physiological relevance of VDR down-regulation may not be related to P homeostasis rather than to Ca homeostasis by minimizing the hypercalcemia induced by P depletion. PMID- 7495174 TI - Parish nursing in Alberta. PMID- 7495175 TI - Margaret Scott Wright research day: Parish Nursing Symposium March 14, 1995. PMID- 7495176 TI - Market research: an essential part of independent practice. PMID- 7495177 TI - Health care reorganization in Alberta: key elements of program management. PMID- 7495178 TI - On my mind: the coming 'jobshift' in nursing. PMID- 7495179 TI - A new nurse registration examination for Canadian nurses. PMID- 7495181 TI - The rational management of epilepsy through greater understanding of the disease. May 27-28, 1994, Budapest. Proceedings. PMID- 7495180 TI - Health care reorganization in Alberta: nurses working successfully in a program structure. PMID- 7495182 TI - Latest advances in epilepsy surgery. AB - Recent advances in epilepsy surgery are a result of improved methods of assessment and diagnosis, a better understanding of seizures, the possibility of surgery at a younger age and the development of new surgical techniques. These factors have led to a wider selection of candidates for epilepsy surgery and shorter treatment trials with anti-epilepsy drugs before surgery is considered. The psychosocial indications for surgery, however, are often not examined thoroughly enough. Epilepsy surgery shows the best results following temporal lobe excision, with 68% of patients becoming seizure-free and 24% showing an improvement. Extra-temporal surgery results in 45% of patients seizure-free and 35% improved. The results of epilepsy surgery in children are similar to those of adults, with 67% of children becoming seizure-free and 21% showing improvement, following successful extensive cortical excision for non-inflammatory lesions. The beneficial psychosocial-economic effects of epilepsy surgery are, however, seldom documented and more research is needed into methods of quantifying, in broader perspectives, the outcome of surgery. PMID- 7495183 TI - Recent developments in the epidemiology of epilepsy. AB - Epidemiological incidence data are important to determine those who are at risk of epilepsy and to understand the aetiology of the condition. A number of studies have been reported recently, which allow comparisons across populations. There appears to be geographical variation in the incidence of epilepsy, with a higher incidence in rural South America than in developed countries: no large scale incidence studies have yet been carried out in Africa or Asia. A consistent pattern suggesting that the onset of epilepsy occurs at the extremes of life has also been shown. Although a number of factors are often identified as causes of epilepsy, studies have identified a definitive aetiology in only one-third of all newly diagnosed cases. Epidemiological studies have, however, confirmed the importance of postnatal insults and quantified the risk. Family history may also be considered a risk factor for epilepsy in epidemiological terms. PMID- 7495184 TI - Prevention of neuronal cell death by anticonvulsants in experimental epilepsy (extended abstract). AB - In human temporal lobe epilepsy, neuronal cell damage is found in the hippocampus (1) as well as in the amygdaloid complex (2) and entorhinal cortex (3). Data from animal studies show that recurrent seizures selectively injure certain populations of neurons in the temporal lobe structures (4). Whether or not recurrent seizures cause neuronal damage in human epilepsy is still under investigation. A new challenge for anti-epilepsy medication is not only to suppress clinical seizures but also to prevent any neuronal damage caused by the seizures. PMID- 7495185 TI - Management of epilepsy in Hungary. AB - The management of epilepsy in Hungary is becoming a science in its own right (epileptology) and clinicians are conforming to international standards of practice. In this paper, the historical background of epileptology and the organization of treatment in Hungary are reviewed, and the current state of epidemiological data, development of diagnostic techniques and pharmacological and surgical therapy summarized. Particular emphasis is placed on: new strategies in morphological and functional neuroimaging; up-to-date electrophysiological diagnosis; clinical use of recently introduced pharmacological regimens. The results from questionnaires about public awareness and attitudes towards epilepsy, and quality of life in Hungarians with epilepsy are presented. PMID- 7495186 TI - Vigabatrin: effect on brain GABA levels measured by nuclear magnetic resonance spectroscopy. AB - Vigabatrin is undoubtedly one of the most exciting anti-epilepsy drugs in use today. Many open and controlled clinical trials have confirmed that it is particularly effective in controlling partial epileptic seizures with or without secondary generalization. Vigabatrin acts to increase GABA levels in the presynaptic nerve terminal by inhibiting the activity of GABA-transaminase. There is no direct correlation between the blood or brain concentration of vigabatrin and its clinical effect, so monitoring vigabatrin levels is not predictive of patient response. However, it is possible to relate the activity of vigabatrin to levels of GABA in the brain, measured by nuclear magnetic resonance spectroscopy (NMRS). NMRS studies show that following administration of vigabatrin, brain concentrations of GABA rise to about 2-3 times their baseline values. This 'extra' GABA is held within the nerve terminal, and is only released during synaptic transmission. Although there appears to be a clear dose-response relationship up to 3 g/day, it is not well documented if higher doses result in proportionately higher brain GABA levels. This finding seems to support the results of clinical studies suggesting that the optimal dose of vigabatrin may be 3 g/day. There is also some evidence for a correlation between the concentration of GABA in the brain and the clinical outcome. Continuing investigations using NMRS aim to confirm these preliminary findings, and to determine the time course and extent of changes in brain GABA levels after vigabatrin administration. PMID- 7495187 TI - Pharmacological principles as a basis for polytherapy. AB - Most patients with newly diagnosed epilepsy can be optimally controlled by prescribing a single anti-epilepsy drug, selected on the basis of its efficacy and safety profile. In about one-third of patients, however, seizures persist during monotherapy, despite the intake of the maximally tolerated drug dose. In such cases, substantial therapeutic benefit may be achieved by prescribing appropriate drug combinations. Safe use of multiple drug therapy requires a good knowledge of clinical pharmacology, particularly an awareness of potentially adverse drug interactions. As many older anti-epilepsy drugs have similar modes of action, their interaction may not always be of clinical benefit, because drug side-effects may also be additive. There is, however, evidence that specific combinations may be particularly advantageous; for example, valproate and ethosuximide in the management of refractory absence seizures. Compared with older drugs, some of the recently developed agents possess different and more selective mechanisms of action, which may result in enhanced therapeutic benefit when specific combinations are used. Preliminary observations do suggest that, in some cases, the efficacy exhibited by certain new drugs could be explained in terms of their pharmacological effect being 'complementary' to that of concurrently used agents. PMID- 7495188 TI - Rationalized polytherapy for epilepsy. AB - In the past, epilepsy was usually treated with polytherapy, but with little knowledge as to the interactions and side-effects of the combinations of the anti epileptic drugs used. Adverse events and sparse clinical knowledge led to monotherapy becoming the treatment regime of choice. A new generation of drugs, which are well-tolerated and have few or predictable interactions, have enabled the reassessment of polytherapy for the treatment of epilepsy. Extensive clinical trials of these drugs are allowing the emergence of a new, rationalized approach to polytherapy. In our study, 19 patients with refractory partial epilepsy, and who were 'socially active and integrated into society', received vigabatrin as add-on therapy. Patients were taking a mean of 1.5 drugs, and five patients were taking small doses of drugs which lead to tolerance, such as barbiturates and benzodiazepines. With vigabatrin as add-on therapy, 14 patients (73%) had a greater than 50% reduction in seizure frequency, and 10 (52%) had a greater than 70% reduction in seizure frequency. In one patient, seizure frequency increased, and two patients developed myoclonic jerks. Vigabatrin was not shown to have any harmful effects in extensive laboratory, EEG and cognitive function tests. In fact, a minor improvement occurred in visual memory, which was probably related to the reduction in seizures. Addition of vigabatrin may, therefore, be of benefit to patients with partial epilepsy refractory to monotherapy with standard anti-epilepsy drugs. PMID- 7495189 TI - Vigabatrin as first add-on treatment in carbamazepine-resistant epilepsy patients. AB - Numerous clinical reports and several controlled clinical trials have confirmed that vigabatrin is both effective and well-tolerated as an add-on treatment for patients with drug-resistant epilepsy. This report presents the results of a study of 40 patients (22 women and 18 men), aged 19-60 years (mean 37 years), with partial seizures (with or without secondary generalization) and receiving carbamazepine, 600-1800 mg/day. Vigabatrin was given as first add-on drug at a dose of 2-3 g/day for an average of 6 months, in order to assess the clinical response before considering other anti-epilepsy drugs. There was a significant decrease in seizure frequency, from a median of 13 seizures/month at baseline, to 3 seizures/month during the last month on vigabatrin (p < 0.01). Seven patients became seizure-free (17.5%). The most common adverse events experienced during the study were drowsiness, diplopia/blurred vision, and were already present before vigabatrin treatment. In conclusion, vigabatrin is effective as a first add-on therapy for partial epilepsy, refractory to carbamazepine monotherapy, and appears to be a worthy clinical alternative to other drug combinations. PMID- 7495190 TI - Pharmacokinetic and pharmacodynamic drug interactions during treatment with vigabatrin. AB - Pharmacokinetic drug interactions take place when one drug interacts with another at the level of metabolism, absorption or excretion. Pharmacodynamic interactions take place at the level of receptor sites, where they may have additive or potentiating effects. Vigabatrin is relatively free of pharmacokinetic interactions, and though it is associated with about a 20% decrease in serum levels of concomitantly administered phenytoin, the reduction is of little clinical significance. The mechanism underlying this effect is unknown. Because vigabatrin increases GABA-mediated inhibition in the brain (an action that is believed to account for its anticonvulsant effects), it might be expected to potentiate the CNS effects of benzodiazepines and alcohol. However, very sensitive eye movement studies have failed to detect any evidence of such an interaction. Overall, vigabatrin appears to be remarkably free of drug interactions. As a result, it is easier to use in clinical practice than older anti-epilepsy agents. Perhaps the most important finding of the interaction studies with vigabatrin is that there is no need for patients receiving the drug to be told to avoid alcohol. PMID- 7495191 TI - Long-term efficacy and cognitive effects of vigabatrin. AB - Vigabatrin is effective as add-on therapy in about 50% of patients with partial epilepsy refractory to drugs. Furthermore, at least half of the original responders maintain the response over several years. As monotherapy, both vigabatrin and carbamazepine seem to be successful in a similar proportion of newly diagnosed patients with epilepsy, but carbamazepine monotherapy fails more often due to side-effects and vigabatrin more often due to lack of efficacy. However, vigabatrin monotherapy seems to be extremely well tolerated, particularly in relation to cognitive function. PMID- 7495192 TI - Behavioural abnormalities and retention rates of anti-epilepsy drugs during long term treatment of epilepsy: a clinical perspective. AB - Vigabatrin is among the most promising of the new anti-epilepsy drugs, but two unrelated complications have been noted in patients receiving vigabatrin for chronic refractory epilepsy. These are, transient behavioural abnormalities and falling numbers of patients continuing to take vigabatrin (decreasing retention rate) during long-term treatment. Psychotic reactions have been carefully documented both in patients receiving standard and new anti-epilepsy drugs, and in up to 6% of patients after epilepsy surgery. As many psychotic episodes are associated with a dramatic cessation of seizures (i.e. high efficacy of treatment), it is not surprising that behavioural abnormalities are more often seen in patients receiving higher, more effective doses of vigabatrin, particularly those taking more than 3 g/day. A gradual dose increase and slow withdrawal is recommended for possible prevention of behavioural abnormalities, at least in some patients. During treatment with standard anti-epilepsy drugs, approximately 50% of patients remain on randomized treatment with the same drug for several years. For oxcarbazepine, a new drug, the figure is 60%, and for vigabatrin it is about 50%. Although comparative trials are not available, the retention rate for vigabatrin does not appear to be strikingly different to that reported for conventional anti-epilepsy drugs. In summary, behavioural abnormalities and falling retention rates appear to be general features of chronic epilepsy, and are certainly not exclusively associated with the use of vigabatrin. Controlled trials are required to establish the specific contribution, if any, of vigabatrin in development of these complications of intractable epilepsy. PMID- 7495193 TI - Viral respiratory infections and their role as public health problem in tropical countries (review). AB - Acute respiratory infections (ARI) are a major cause of morbidity and mortality throughout the world. Data from the World Health Organization indicate that there are at least 2.2 million deaths from ARI throughout the world each year. A considerable number of study have been performed in different countries to assess the etiological role of viruses in ARI and is now clear that the majority of infections of the respiratory tract are caused by viruses. In tropical countries information on the viral etiological agents of ARI is rather scanty. Nevertheless data from Papua New Guinea, Polynesia, India, Pakistan and Singapor show that influenza occurs frequently in tropical countries. The other respiratory viruses especially respiratory syncytial virus, parainfluenza viruses and adenoviruses also play significant role as etiological agents in many tropical countries as Panama, Jamaica, Brazil, Colombia, Trinidad, Uganda, India and Nigeria. The data concerning seasonal prevalence of viral ARI in tropics are contradictory. PMID- 7495194 TI - A low birthweight survivor of perforated necrotizing enterocolitis--a case report. PMID- 7495195 TI - Pregnancy associated alpha-glycoprotein (SP-III) in Nigerian pregnant women. AB - Pregnancy associated protein (SP-III) was measured by rocket electroimmunoassay in the sera of Nigerian women with normal and abnormal singleton pregnancy. A tendency to increases of SP-III levels with advancing gestational age was observed. SP-III may contribute to immunosuppression in preventing anti-allograft responsiveness and levels may also reflect the effect of infections since SP-III is also leucocyte derived. Values in PET and anaemia in pregnancy were similar to values in normal pregnant women. PMID- 7495196 TI - Immunoglobulin levels in malaria infected Nigerians with and without abnormal haemoglobin. AB - Comparative studies were made between malarial parasitaemia in Nigerians with and without abnormal haemoglobins. The three main classes of immunoglobulins (i.e. IgG, A and M) were assayed in these groups of patients and the mean values were compared. Those with abnormal haemoglobins S or C (HbS or HbC) were compared with those with normal control haemoglobin A (HbA). HbSS malarial patients have the highest mean values of the 3 classes of immunoglobulins. This is followed by HbAS patients while patients with normal Hb have lowest mean values for IgG and IgM. The significance of the results is discussed. PMID- 7495197 TI - Teenage prostitution--child abuse: a survey of the Ilorin situation. AB - In the light of civilization that has come to erode some of the sexual constraints in the traditional African society's opennes today toward sexuality and sexual expression, adolescents commence sexual activity at an increasing earlier age and the average marriage come much later. It is therefore not surprising that an increasing proportion of adolescents in Nigeria are engaging in teenage prostitution. This study is an attempt to conduct a general survey into the nature of teenage prostitution in Nigeria and to provide information on the prevalence, factors, trends and characteristics of teenage prostitution. A semi-structured questionnaire was administered to 150 known teenage prostitutes. Among other things the finding suggest that childhood sexual abuse perhaps leads to prostitution. Half of the sample used in the study had their first sexual experience early in life, and about the same number lived with their parents when they began prostituting. Implications of the findings were discussed and suggestions made for further research. PMID- 7495198 TI - Renal vein thrombosis in patients with nephrotic syndrome: CT diagnosis. AB - Two male patients, an adult and a child, with the nephrotic syndrome were recently seen with clinical manifestations of acute Renal Vein Thrombosis (RVT) and the diagnosis was confirmed by Computed Tomography (CT) findings. The adult patient had left RVT, while the child had, at the first presentation, right RVT, followed two months later by the development of a left RVT. Subsequently, twelve asymptomatic patients with nephrotic syndrome were screened for RVT. Two adult male patients were discovered to have right RVT. All the four patients with RVT had associated thrombi in the Inferior Vena Cava (IVC). CT scans showed in all, widening of the involved renal vein containing thrombus, renal enlargement and thrombus in the IVC. CT appears to offer a direct non-invasive method of visualization of the IVC and renal veins, detecting thrombus within these vessels and demonstrating the resulting renal functional and morphological consequences, making it, suitable also for screening an asymptomatic population at high risk for RVT. PMID- 7495199 TI - Decrease in platelet survival and total platelet sialic acid concentration in rats infected with Plasmodium bergei bergei. AB - Suckling Wistar rats aged 3-5 weeks were infected through their dorsal tail vein with P. berghei berghei passed in Swiss albino mice. Platelet recovery and platelet survival using 51Cr-labelled heterologous platelets obtained from adult Wistar rats were determined in the infected animals on different post-infection days and on a group of non-infected rats as controls. Total platelet sialic acid was also determined in the same groups of animals. The results showed reduced platelet recovery, shortened survival and reduced total platelet sialic acid content in the infected animals compared with control values. The reduction in total platelet sialic acid content was related to the degree of parasitaemia and reached significant levels on the 5th post-infection day. It is concluded that the shortened platelet survival and reduced total platelet sialic acid content observed in the P. berghei infected rats were causally related and may account for the thrombocytopaenia reported in experimental and natural malaria infections of animals and man. PMID- 7495200 TI - Respiratory syncytial virus--associated lower respiratory diseases in hospitalised pre-school children in Ibadan. AB - An analysis of the features of lower respiratory diseases (LRD) associated with Respiratory Syncytial Virus (RSV) in 21 children aged 1-23 months, indicated that the clinical features did not differ appreciably from those described for LRD due to other infective agents. It was however, remarkable that none of the children was grossly malnourished, neither was there any death. The final respiratory diagnoses were bronchiolitis alone (6 cases), bronchopneumonia (6 cases), empyema/pleural effusion (3 cases, including one with bronchopneumonia), lobar pneumonia (3 cases, including one with associated bronchopneumonia). Streptococcus pneumonia and Klebsiella pneumoniae respectively, were isolated from blood culture in the two cases with lobar pneumonia alone. One child had laryngotracheobronchitis (croup) alone and another, croup associated with bronchiolitis and bronchopneumonia. Radiologically, the single most common lesion was patchy consolidation (8 cases) followed by hyper-inflation (6 cases); in one case, there was no observable lesion on the chest radiograph. All the 6 children who had bronchiolitis alone were aged 12 months and below, and were well nourished; conversely, 8 of the 12 children with pneumonia alone or in addition to other lesions and both children with pleural effusion, were older than 12 months and underweight. Other viral agents were identified in 8 (38%) of the cases. Complications included heart failure in 4 cases and a combination of anaemia and heart failure in one child with empyema. In a child with symptom complex and radiological features suggestive of acute obstructive lower respiratory infection with immunofluorescence evidence of RSV infection, age and nutritional status are important factors in determining the need for antibiotic therapy pending the outcome of bacterial cultures. PMID- 7495201 TI - Hyponatraemia, birthweight and neonatal jaundice. AB - Cord serum sodium levels were estimated in 96 full term singletons delivered by spontaneous vertex delivery. Thirty-two of these infants had cord serum sodium of less than 130 mmol/L and 64 had cord serum sodium of > or = 130 mmol/L. Serum bilirubin estimated on day 4 of life of the babies demonstrated mean unconjugated bilirubin levels of 105.7 mmol/L (S.D.:87.04) and 89.4 mmol/L (S.D.:66.18) in the hyponatraemic and normonatraemic groups respectively (P < 0.05). The study also demonstrated a higher cord sodium and lower serum unconjugated bilirubin in those babies whose mothers did not receive parenteral fluids. It is suggested that hyponatraemia should be excluded in the aetiology of neonatal jaundice. PMID- 7495202 TI - Performances of copper T 380A and multiload copper 375/250 intrauterine contraceptive devices in a comparative clinical trial. AB - An evaluation of the performances of copper T 380A (TCU 380A) and multiload copper 375/250 (MLCU 375/250) intrauterine contraceptive devices (IUCDS) was carried out at University College Hospital, Ibadan, Nigeria (UCH) in a clinical comparative trial. The IUCDS showed similar low vent rates at one year of continuous use. The difference in cumulative net probabilities for termination due to pelvic inflammatory disease was weakly significant at the sixth month of follow up with MLCU 250 having the highest rate. (TCU 380A-0%; MLCU 375-0%; MLCU 250-3.1%; X2 = 6.0; P < 0.05). This significant difference disappeared by the twelfth month of continuous use. Likewise, the difference in cumulative net probabilities for overall termination was significant at six months and insignificant at twelve months of follow up. The continuation rate after one year were 86%, 92% and 87% respectively for TCU 380A, MLCU 375 and MLCU 250. These rates were higher than 80% and 51% quoted for the previously available Lippes loop in the same environment [1,2]. It was therefore concluded that the three IUCDS are comparable in performances in the first year of use and could be used at our family planning clinic or any other clinic in a similar setting. PMID- 7495203 TI - The antipruritic effects of chlorpheniramine, cyproheptadine and sulphapyridine monitored with limb activity meters on chloroquine induced pruritus among patients with malaria. AB - Limb activity meters, otherwise modified self-winding watches that can record limb agitative movements such as in itch-provoked scratch, were introduced for an objective evaluation of the relative effectiveness of three antipruritic drugs: chlorpheniramine, cyproheptadine, and sulphapyridine for palliating pruritus associated with chloroquine chemosuppressive treatment of acute malarial febrile paroxysms in eighteen adult patients. Six fit and healthy subjects were also studied to obtain data for unmedicated controls. The meters were used to monitor the upper and lower limb activities of the patients during nocturnal sleep for 6 hours over 3 consecutive nights, after they had developed the chloroquine-induced pruritus and were then administered the antipruritic medications, six patients per drug, by a random selection. Sulphapyridine antipruritic treatment significantly reduced the activities of the upper limbs of itchy patients much greater than did cyproheptadine (P < 0.0001, right hand; P < 0.01 left hand). However sulphapyridine-treated patients still itched significantly more than controls from the greater activities in the dominant right hand of the patients (P < 0.05). Cyproheptadine had a marginally-better performance than chlorpheniramine, generally, in palliating the chloroquine-induced pruritus but only in one of 3 nights, for the right hand recordings, were the limb activities significantly different. There was no significant difference observed in the activities of the lower limbs for the unmedicated controls compared to itchy patients, irrespective of the antipruritic treatment mode. Five out of the 6 patients treated with sulphapyridine also complained of anorexia plus a feeling of fullness or indigestion.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7495204 TI - Autopsy survey for Alzheimer's disease in Nigerian Africans: a preliminary report. AB - We carried out an autopsy survey on 198 brains of Nigerians aged 40 years and above to determine the occurrence of (neuropathognomonic) changes of Alzheimer's disease (AD) in our elderly patients. Forty five patients (23%) were above 65 years of age. Appropriately stained histological sections of various parts of the brains showed mild cortical neuronal loss and absence of neurofibrillary tangles, senile plaques and amyloid angiopathy--hallmarks of Alzheimer's disease and ageing reported in the Caucasians and Japanese. PMID- 7495205 TI - Evaluation of the "one-minute" test for detecting Helicobacter (Campylobacter) pylori infection. AB - The "one-minute" urease test to detect Helicobacter (Campylobacter) pylori infection was evaluated using histology and culture as the "gold standard". The test was performed in a blinded manner and compared with the conventional Christensen's urease test. Helicobacter pylori was detected in 88 of 100 consecutive patients attending the gastrointestinal clinic for upper endoscopy. Although the "one-minute" urease test was more sensitive [86% (76/88)] than the conventional Christensen's urease test [70% (62/88)], this difference was not statistically significant (P = 0.22). Histology was the most sensitive [97% (85/88)] whilst culture was 80% (70/88) sensitive. All tests exhibited specifications of 100%. The "one-minute" urease test is a simple, rapid and highly specific test to detect Helicobacter pylori which can be performed at endoscopy. PMID- 7495206 TI - An ethnographic study of acute respiratory infections in four local government areas of Nigeria. AB - An ethnographic study was conducted in four local government areas of Nigeria. The techniques of informal unstructured interviews and participant observation were used. A total of 104 focus group discussions with 53 groups of mothers, 21 groups of grandmothers, and 30 groups of fathers were conducted. Perception of causes of ARI ranged from cold water, to heredity, poor hygiene, exposure to smoke and dust and the supernatural forces. Preventive measures described were related to the perceived causes. For those groups that discussed home remedies to the treatment of ARI, the remedies described for cough included herbal drinks (39% of groups); honey with lemon (19.5%); eating specific vegetables believed to relieve cough (8.4%); and preparations containing palm oil (21.7%). Remedies described for measles included herbal drinks (62%); local tropical creams (24%); and palm wine (13.7%). Those for ear infections included drops of herbal mixtures in the ear (29.4%); putting various type of oil in the ear (38%); plugging the ear with cotton wool previously dipped in honey, or alcohol (17%). The findings of this study have implications for the Health Education Component of the National ARI Control Programm which Nigeria recently embarked upon. There is also the need for research on the efficacy and any possible adverse effects of identified home remedies. PMID- 7495207 TI - Complement and immunoglobulin levels in Ilorin Nigeria, and environ. AB - The levels of complement components C3 and C4 were determined in EDTA (ethylene diamine-tetra-acetic acid) plasma of 106 apparently healthy people aged 11-50 years, and in 23 paired maternal and cord plasma in Ilorin. The immunoglobulins G,A and M concentrations of these paired samples were also assayed. Furthermore, serum IgG, IgA and IgM levels were quantitated in 501 apparently healthy residents of Ilorin (1-84 years old, n = 313) and its suburban community, Shao (11-80 years old, n = 188). Generally, there were no significant sex-related differences in C3 and C4 values, but these values increased steadily with age. C3 and C4 levels in cord plasma were high, representing 73% and 88% of adult levels respectively. Maternal levels of these complement components were higher (P < 0.02) than values obtained in non-pregnant females of comparable age. Immunoglobulin G values obtained for maternal and cord samples compare well with figures reported previously from other centres in Nigeria. Only 1 of 23 IgA and 2 of 23 IgM samples showed detectable values of these immunoglobulins in cord blood. The mean concentrations of immunoglobulins obtained were similar for IgG and lower for IgA and IgM in Ilorin, but higher for IgG and lower for IgA and IgM in Shao, when compared with data reported from Enugu and Zaria--both in Nigeria. Shao showed significantly higher mean level of IgG but similar levels of IgA and IgM to values obtained in Ilorin. Generally, IgG and IgA concentrations rose gradually up to age group 31-40 and 41-50 years respectively; and then declined. (ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7495208 TI - Disproportionate intraventricular septal hypertrophy in Nigerians with essential hypertension. AB - It is not clear how frequently hypertrophic cardiomyopathy coexists with essential hypertension or whether chronic hypertension per se, induces hypertrophic cardiomyopathy (hypertensive hypertrophic cardiomyopathy). Eighty three Nigerians with essential hypertension alone underwent M-mode and 2 dimensional echocardiographic study. Twenty per cent (17 patients) had disproportionate intraventricular septal hypertrophy (septum: posterior wall ratio > or = 1.4). These subgroup tended to be elderly, with average age of 57.7 years, and were predominantly male (70%). Two patients in this subgroup (2.4% of total) additionally had thickened posterior left ventricles and a hyperdynamic left ventricular systolic function (fractional fibre shortening of > or = 45%). This picture is consistent with "hypertensive hypertrophic cardiomyopathy". Routine echocardiography may permit the early identification and thus a closer scrutiny and follow up of essential hypertensive patients with disproportionate septal thickness. The prognostic implications of this requires long term study. PMID- 7495209 TI - Comparison of single serum progesterone and endometrial biopsy for confirmation of ovulation in infertile Nigerian women. AB - Infertility remains a medico-social problem in Nigeria and it accounts for a large percentage of outpatient gynecological consultations. The evaluation of the infertile couple remains a continuing challenge to the practising doctor in this part of the world. The need to evaluate the two methods commonly used for determining ovulation in these patients is indicated. Endometrial biopsy specimen and a single sample for serum progesterone estimation were obtained simultaneously in the luteal phase from 50 normally menstruating infertile Nigerian women. Subsequent analysis showed that a serum progesterone value of 6.6 nmol/l (2.2 ng/ml) or above was always associated with a secretory endometrium. Forty-six cycles yielded sufficient information to compare the two methods for confirmation of ovulation. Patients who ovulated with a progesterone value of 6.6 nmol/l (2.2 ng/ml) were 91.3% (42/46) or above, while 89% (41/46) showed secretory endometrium. Forty-six of the cases 86.9% (40/46) were judged to have ovulated by both parameters while 6.5% demonstrated anovulatory cycle using both criteria. From the study, a significant correlation was obtained between endometrial biopsy and progesterone assay methods in confirming ovulation. PMID- 7495210 TI - A symposium: progress in hypertensive heart disease: microangiopathy, molecular and cellular biology, cardiac arrhythmias, and heart failure. February 23-24, 1995, Bilbao, Spain. Proceedings. PMID- 7495212 TI - The cardiac renin-angiotensin-aldosterone system and hypertensive cardiac hypertrophy. AB - Angiotensin-converting enzyme inhibitors have proven to be uniquely effective in inducing regression, or preventing the occurrence, of ventricular hypertrophy associated with systemic hypertension. This has pointed, for many years, to a possible direct involvement of the renin-angiotensin system in the pathogenesis of cardiac hypertrophy. Over the last 10 years further supporting evidence has been forthcoming about direct trophic effects of angiotensin II in several experimental systems. Additionally, we now have rather conclusive evidence for the existence of a local, intracardiac renin-angiotensin system, which is capable of synthesis of all components of the system, and of cleaving, via the classic pathway, angiotensin peptides from the precursor, angiotensinogen. Moreover, a number of studies have demonstrated the capacity of regulatory response and modulation of activity of the local system in response to a variety of pharmacologic perturbations as well as differential expression of specific components under pathologic conditions, including compensatory hypertrophy and remodeling after myocardial infarction, pressure overload hypertrophy, and volume overload hypertrophy. Continued research into the role of the cardiac renin angiotensin system in the pathogenesis of cardiac hypertrophy and failure will provide us with the tools to devise more specific, targeted strategies for therapeutic intervention or prevention. PMID- 7495211 TI - Toward the biochemical assessment of myocardial fibrosis in hypertensive patients. AB - The serum concentrations of amino-terminal procollagen type III and carboxy terminal procollagen type I-derived peptides, which have been proposed as useful markers of the tissue synthesis of collagen types III and type I, respectively, were abnormally increased in patients with essential hypertension and became normal after angiotensin-converting enzyme (ACE) inhibition. An association was found between baseline serum concentrations of these peptides and left ventricular hypertrophy, diastolic dysfunction, and ventricular arrhythmias in hypertensive patients. On the other hand, increased serum concentration of the carboxy-terminal procollagen type I-derived peptide was found in spontaneously hypertensive rats compared with normotensive Wistar-Kyoto control rats. An association was found between the serum concentration of this peptide and the extent of myocardial fibrosis and the hydroxyproline concentration in the left ventricle of spontaneously hypertensive rats. It is proposed that procollagen derived peptides in serum may be markers of exaggerated collagen tissue synthesis involved in hypertensive myocardial fibrosis. PMID- 7495213 TI - Molecular and cellular biology of the senescent hypertrophied and failing heart. AB - During aging, experimental studies have revealed various cellular changes, principal among which is myocyte hypertrophy, which compensates for the loss of myocytes and is associated with fibrosis. The expression of alpha-myosin heavy chain is replaced by that of the isogene beta-myosin, which leads to decreased myosin adenosine triphosphatase (ATPase) activity. In consequence, contraction is slower and more energetically economical. The Ca(2+)-ATPase of the sarcoplasmic reticulum and Na+/Ca2+ exchange activity are decreased, which probably explains the reduced velocity of relaxation. Membrane receptors are also modified, since the density of both the total beta-adrenergic and muscarinic receptors is decreased. The senescent heart is able to hypertrophy in response to overload and to adapt to the new requirements. Similar alterations are observed both in the senescent heart and in the overloaded heart, in clinical as well as in experimental studies; however, differences do exist, especially in terms of fibrosis and arrhythmias. PMID- 7495214 TI - Response of hypertensive left ventricular hypertrophy and coronary microvascular disease to calcium antagonists. AB - Hypertensive left ventricular hypertrophy comprises not only myocyte hypertrophy, but is often associated with interstitial fibrosis and structural alterations of the coronary microcirculation. The consequences are early diastolic dysfunction and often later systolic dysfunction of the left ventricle, resulting in congestive heart failure. Involvement of the coronary resistance vessels leads to an impairment of coronary flow reserve despite normal epicardial arteries. Therefore, antihypertensive treatment should aim at reversing myocyte hypertrophy, restoring myocardial structure, and improvement in coronary flow reserve apart from blood pressure lowering. Many clinical studies have shown that calcium antagonists are effective in lowering blood pressure and can induce regression of left ventricular hypertrophy. Moreover, experimental studies have shown a restoration of myocardial structure. Recent clinical studies have demonstrated a marked improvement of the impaired coronary vasodilator reserve in hypertensive patients after long-term treatment with calcium antagonists. In summary, calcium antagonists can be regarded as an antihypertensive treatment modality that is able to restore myocardial structure and to repair coronary microcirculation and therefore can be considered as causative treatment of hypertensive cardiac remodeling. PMID- 7495215 TI - Microvascular angina in systemic hypertension: diagnosis and treatment with enalapril. AB - The causes of hypertensive microvascular ischemia are reviewed along with diagnostic factors. Stress/rest thallium-201 scintigraphy is shown to have a predictive value of 78% for a diagnosis of microvascular disease in hypertensive patients with exertional angina and left ventricular hypertrophy. Lack of isotope uptake at peak stress correlates well with the decrease in coronary flow reserve in ischemic segments, which is 2-3 times lower than in normal subjects. Treatment with enalapril produces regression of left ventricular hypertrophy, normalization of thallium-201 uptake, and an increase in exercise capacity in patients with microvascular angina. PMID- 7495216 TI - Angina pectoris and normal coronary arteriograms: clinical presentation and hemodynamic characteristics. AB - Up to 30% of patients undergoing coronary angiography for the assessment of chest pain suggestive of coronary artery disease have "normal" studies. Several reports have indicated that a proportion of patients with angina and normal coronary arteriograms have reduced coronary flow reserve. The interpretation of these findings is, however, controversial as the majority of patients do not have definitive evidence for myocardial ischemia and have a good long-term prognosis. The clinical presentation of patients with angina and normal coronary arteriograms differs in different series and this may be just a reflection of the heterogeneous nature of the syndrome. A diversity of pathogenetic mechanisms have been postulated to explain "syndrome X" (chest pain and normal coronary arteriograms) but little is known at present about the true nature of the syndrome. The present article discusses the clinical and hemodynamic features of this intriguing disorder with particular reference to patients with syndrome X and microvascular angina. PMID- 7495217 TI - Congestive heart failure due to hypertensive ventricular diastolic dysfunction. AB - Left ventricular (LV) diastolic dysfunction is the first discernible manifestation of heart disease in hypertensive patients. Arterial hypertension with LV hypertrophy leads to reduced preload followed by impaired cardiac output (systolic dysfunction stemming from primary diastolic dysfunction). Diastolic dysfunction leads more often than systolic dysfunction to hypertensive heart failure and is in many cases clearly distinguishable from heart failure with low ejection fraction (EF). Mortality due to heart failure from impaired inotropism is higher than mortality due to diastolic dysfunction, but morbidity is lower. Hypertensive cardiomyopathies can be divided into 4 ascending categories, according to the pathophysiologic and clinical impact of hypertension on the heart: Degree I: LV diastolic dysfunction with no associated LV hypertrophy Degree II: LV diastolic dysfunction with echocardiographic LV hypertrophy Degree IIA: Normal exercise capacity in terms of maximal oxygen consumption Degree IIB: Impaired exercise capacity in terms of maximal oxygen consumption Degree III: Congestive heart failure (severe dyspnea and radiographically determined pulmonary edema with normal (> or = 50%) EF Degree IIIA: LV mass/volume ratio > 1.8 with little or no myocardial ischemia Degree IIIB: LV mass/volume ratio < 1.8 with significant myocardial ischemia Degree IV: Profile of dilated cardiomyopathy; LV hypertrophy and impaired EF (< 50%). PMID- 7495218 TI - Systolic ventricular dysfunction and heart failure due to coronary microangiopathy in hypertensive heart disease. AB - Left ventricular hypertrophy in arterial hypertension is characterized by myocyte hypertrophy, myocardial fibrosis, and structural changes of the intramural coronary arteries. Hypertensives with or without left ventricular hypertrophy have a reduced coronary vasodilator reserve due to alterations of the coronary microcirculation. The impairment in coronary vasodilator reserve is likely to initiate a process of malperfusion and malnutrition concomitant with increased metabolic demands. Further, malperfusion is supported by an increase in diastolic filling pressure, which will enhance the extravascular component of coronary resistance. The sum of interactions of these structural alterations of myocardium, interstitium, and coronary vasculature are likely to initiate and maintain a process of myocardial malperfusion and malnutrition, which can provoke functional depression of the myocardial performance, a loss of contractile proteins, an increase in interstitial fibrosis, and, not least, an overall decrease in contractile function in long-standing cardiac hypertrophy. Finally, the reversal of these processes by adequate antihypertensive treatment may contribute to renormalization of cardiac function and to prevention of late cardiac failure in hypertensive heart disease. PMID- 7495219 TI - Cardiac arrhythmias and left ventricular hypertrophy in systemic hypertension and their influences on prognosis. AB - Left ventricular hypertrophy (LVH) is the adaptative mechanism of the heart to systolic overload of the left ventricle. Nevertheless, LVH plays a role in some complications, such as cardiac arrhythmias. Patients with LVH are more likely to develop ventricular arrhythmias than the hypertensive population without LVH. Further, the relation between left ventricular mass and ventricular arrhythmias is graded and continuous. The arrhythmias described in hypertensive patients with LVH are usually isolated premature ventricular contractions. The presence of electrocardiographic criteria of LVH represents a risk of higher incidence of sudden death, especially in men. The risk is even greater in the presence of ventricular arrhythmias. The presence of late potentials has been recently characterized as more related to ventricular arrhythmias than LVH. Antihypertensive drugs that can reduce LVH also have a beneficial effect on cardiovascular morbility and mortality. PMID- 7495220 TI - Evidence favoring the hypothesis that ventricular arrhythmias have prognostic significance in left ventricular hypertrophy secondary to systemic hypertension. AB - In the present review 6 lines of evidence will be discussed that suggest a prognostic significance for ventricular arrhythmias in patients with systemic hypertension and left ventricular hypertrophy: (1) in patients with systemic hypertension there is a statistical relation between asymptomatic ventricular arrhythmias and left ventricular hypertrophy; (2) in nonhypertensive left ventricular hypertrophy the prognostic value of ventricular arrhythmias is well known; (3) left ventricular hypertrophy is related to sudden death in patients with systemic hypertension; (4) it is generally acknowledged that ventricular arrhythmias are a frequent cause of sudden death; (5) there is experimental evidence to support the arrhythmic risk of left ventricular hypertrophy; and (6) it has been recently demonstrated that ventricular arrhythmias influence mortality in patients with left ventricular hypertrophy secondary to systemic hypertension. However, whether asymptomatic ventricular arrhythmias are specific markers for more severe sustained arrhythmias, or just markers for a more severe stage of the disease, remains to be determined. PMID- 7495221 TI - Role of angiotensin II and prostaglandin E2 in regulating cardiac fibroblast collagen turnover. AB - In hypertensive heart disease, after myocardial infarction or in congestive heart failure, myocardial fibrosis presenting as a diffuse perivascular and interstitial accumulation of fibrillar collagens within the normal connective tissue structures of the myocardium is associated with an activated renin angiotensin system (RAS). This reactive fibrosis occurs in the overloaded left ventricle and the nonoverloaded right ventricle irrespective of myocyte necrosis or the development of myocyte hypertrophy. Therefore, it appears that hemodynamic factors or the load of the ventricle are not primarily responsible for the adverse fibrous tissue response in the myocardium, and humoral factors may play a key role in regulating the myocardial collagen matrix. The neurohumoral response in hypertensive heart disease, after myocardial infarction with overall deterioration of left ventricular function or congestive heart failure leads to an activation of either the cardiac or the circulating RAS, which closely interacts with the bradykinin-prostaglandin system. To ascertain whether the RAS modulates collagen fibroblasts that express mRNAs for types I and III collagens (the major fibrillar collagens in the heart) and matrix metalloproteinase 1 (MMP1; the key enzyme for collagen degradation), collagen synthesis was measured by [3H]proline incorporation normalized to total protein synthesis and MMP1 activity was determined by degradation of [14C]collagen in cultured fibroblasts after 24-hour incubation with various concentrations of angiotensin II or PGE2 (10(-11)-10(-3) M) under serum-free conditions. In addition, effects of angiotensin II were evaluated in the presence or absence of either type 1 (ICI D8731) or type 2 (PD 123177) angiotensin II (AT1 or PGE2 (10(-11)-10(-3) M) under serum-free conditions.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7495222 TI - The present status of antioxidant vitamins and beta-carotene. PMID- 7495223 TI - Antioxidant Vitamins and Beta-Carotene in Disease Prevention. Proceedings of a symposium. Berlin, Germany, October 10-12, 1994. PMID- 7495224 TI - Chronic and degenerative disease: major causes of morbidity and death. AB - In OECD (Organization for Economic Cooperation and Development) countries (Australia, Austria, Belgium, Canada, Denmark, Finland, France, Germany, Greece, Ireland, Italy, Japan, Luxembourg, Netherlands, New Zealand, Norway, Portugal, Spain, Sweden, Switzerland, United Kingdom, and United States), mortality at ages 35-69 y has fallen by 38% since 1955 and the risk of dying before age 70 y has been reduced to 23%. In middle and old age, chronic and degenerative diseases are the principal causes of death. Neoplasms, ischemic heart disease, and cerebrovascular disease cause three-quarters of all deaths at 35-69 y of age and two-thirds at older ages. Mortality from vascular disease has declined rapidly. Mortality from neoplasms has not declined rapidly; however, mortality not attributed to smoking has and there is no general increase in cancer attributable to industrial development. Despite lower death rates in nonsmokers than in cigarette smokers, vascular disease and neoplasms remain the targets for prevention. With increased duration of life, conditions that cause serious disability in old age become more important and the principal foci for research. These conditions are also chronic and degenerative. PMID- 7495225 TI - Oxidative stress in acute and chronic pancreatitis. AB - Oxygen radicals mediate an important step in the initiation of acute pancreatitis. These reactive oxygen metabolites are generated at an early stage of the disease. The source of the enhanced production of oxygen radicals, however, still remains unclear. Experimentally, the effectiveness of antioxidant treatment varies from one model to the other, the differences depending on the experimental model and not on the form of pancreatitis that was induced. In most studies, the experimental animals were pretreated before acute pancreatitis was induced. This does not mirror clinical reality because patients are admitted to the hospital after the onset of the disease. It was shown in cerulein-induced pancreatitis, however, that scavenger treatment also mitigated the pancreatic tissue damage after induction of acute pancreatitis. Moreover, antioxidant treatment also attenuated the extrapancreatic complications, thus improving the final outcome of the disease. Initial indirect observations also suggest that in human acute, acute recurrent, and chronic pancreatitis, oxygen free radicals are generated and add to the damage. Concomitantly, these patients suffer from a severe depletion of oxidative stress. Whether or not this disbalance is instrumental in the development and course of disease remains unanswered. Supplementation with antioxidants that are deficient in patients with acute pancreatitis might be a feasible option to the present therapy to avoid extrapancreatic complications. Well-defined, controlled clinical trials involving patients suffering from acute pancreatitis are therefore needed to validate the role of oxygen radicals in this disease. PMID- 7495226 TI - Vitamins E and C, beta-carotene, and other carotenoids as antioxidants. AB - Tocopherols and tocotrienols (vitamin E), ascorbic acid (vitamin C), and the carotenoids react with free radicals, notably peroxyl radicals, and with singlet molecular oxygen (1O2), which is the basis for their function as antioxidants. RRR-alpha-Tocopherol is the major peroxyl radical scavenger in biological lipid phases such as membranes or low-density lipoproteins. Ascorbic acid is present in aqueous compartments (eg, cytosol, plasma, and other body fluids) and can reduce the tocopherol radical; it also has several metabolically important cofactor functions in enzyme reactions, especially hydroxylations. These micronutrients need to be regenerated on oxidation in the biological setting, hence the need for further coupling to nonradical reducing systems such as glutathione-glutathione disulfide, dihydrolipoate-lipoate, or NADPH-NADP+ and NADH-NAD+. Carotenoids, such as beta-carotene, lycopene, and some oxycarotenoids, eg, zeaxanthin and lutein, exert antioxidant functions in lipid phases by quenching 1O2 or free radicals. There are pronounced differences in tissue carotenoid patterns, extending also to the distribution between the all-trans and various cis isomers of the respective carotenoids. Physical quenching leaves the structure intact, so that in this mode the carotenoids do not require a regeneration reaction. PMID- 7495227 TI - Interaction among vitamin C, vitamin E, and beta-carotene. AB - The effects of vitamin C (ascorbic acid), vitamin E (alpha-tocopherol), and beta carotene as antioxidants and their cooperative action against the oxidation of lipid in solution, membranes, and lipoproteins have been studied and reviewed. Ascorbic acid and alpha-tocopherol act as potent, and probably the most important, hydrophilic and lipophilic antioxidants, respectively. They function at their own site individually and furthermore act synergistically. beta-Carotene has lower reactivity toward radicals than does alpha-tocopherol and acts as a weak antioxidant in solution. It is more lipophilic than alpha-tocopherol and is assumed to be present at the interior of membranes or lipoproteins, which enables it to scavenge radicals within the lipophilic compartment more efficiently than does alpha-tocopherol. The cooperative interaction between vitamin C and vitamin E may be quite probable, that of vitamin C and beta-carotene is improbable, whereas that between vitamin E and beta-carotene may be possible. PMID- 7495228 TI - Dietary carotenoids inhibit neoplastic transformation and modulate gene expression in mouse and human cells. AB - Many epidemiologic studies have associated the consumption of diets rich in fruit and green and yellow vegetables with a decreased risk of cancer. Of the many components of such a diet, the content of carotenoids, particularly beta carotene, has been most consistently linked to decreased risk. The biological mechanism for such protection is currently unclear. Multiple possibilities exist: carotenoids are potent antioxidants and oxidative stress is known to contribute to carcinogenesis; many carotenoids can be converted to retinoids, these are known cancer preventive agents at several anatomic sites; and carotenoids may possess additional actions in mammalian cells. In a model in vitro system we showed that carotenoids both with and without provitamin A activity inhibit carcinogen-induced neoplastic transformation, inhibit plasma membrane lipid oxidation, and cause up-regulated expression of connexin 43, a gene coding for the structural unit of a gap junction. This last activity was statistically correlated with the ability to inhibit neoplastic transformation. Activity has also been shown in human cells: in fibroblasts CONNEXIN 43 expression is also up regulated whereas in human keratinocytes grown in organotypic culture beta carotene and canthaxanthin modulate differentiation in a manner qualitatively similar to that of retinoids. These results strongly suggest that carotenoids have intrinsic cancer chemopreventive action in humans. PMID- 7495229 TI - Vitamin E: a sensor and an information transducer of the cell oxidation state. AB - We studied the effects of RRR-alpha-tocopherol and RRR-beta-tocopherol in smooth muscle cells from rat (line A7r5) and human aortas. RRR-alpha-Tocopherol, but not RRR-beta-tocopherol, inhibited smooth muscle cell proliferation in a dose dependent manner at concentrations in the range from 10 to 50 mumol/L. RRR-beta Tocopherol added simultaneously with RRR-alpha-tocopherol prevented growth inhibition. The earliest event brought about by RRR-alpha-tocopherol in the signal transduction cascade controlling receptor-mediated cell growth was the activation of the transcription factor AP-1. RRR-beta-tocopherol alone was without effect but in combination with RRR-alpha-tocopherol prevented the AP-1 activating effect of the latter. Protein kinase C was inhibited by RRR-alpha tocopherol and not by RRR-beta-tocopherol, which also in this case prevented the effect of RRR-alpha-tocopherol. Calyculin A, a protein phosphatase inhibitor, prevented the effect of RRR-alpha-tocopherol on protein kinase C. The data can be rationalized by a model in which a tocopherol-binding protein discriminates between RRR-alpha-tocopherol and RRR-beta-tocopherol and initiates a cascade of events at the level of cell signal transduction that leads to the inhibition of cell proliferation. PMID- 7495230 TI - Determination of optimal vitamin C requirements in humans. AB - Although the recommended dietary allowance provides an estimate for vitamin C ingestion in humans, optimal vitamin C requirements are unknown. We define optimal vitamin C requirements operationally based on the following: dose function relations, availability in the food supply, steady state concentrations in plasma and tissues achieved at each dose of vitamin C, urinary excretion, bioavailability, toxicity, and epidemiologic observations. Optimal vitamin C requirements can be estimated when information is available for at least some of these criteria. PMID- 7495231 TI - Coantioxidants make alpha-tocopherol an efficient antioxidant for low-density lipoprotein. AB - The oxidation of low-density lipoproteins (LDLs) is now commonly implicated as an important early event in atherogenesis. The resulting interest in LDL antioxidation has focused on alpha-tocopherol, the biologically and chemically most active form of vitamin E and quantitatively the major lipid-soluble antioxidant in extracts prepared from human LDL. We review advances made in our understanding of the molecular action of alpha-tocopherol in radical-mediated oxidation of isolated human LDL and how the vitamin's antioxidant activity is enhanced or even dependent on the presence of suitable reducing species, which are referred to as coantioxidants. PMID- 7495232 TI - Epidemiologic evidence for vitamin E in prevention of cardiovascular disease. AB - Ecologic studies of vitamin E have shown that regions with relatively low dietary vitamin E tend to have higher rates of coronary heart disease (CHD), but it is difficult to adjust for other risk factors. Cross-sectional studies in individuals have yielded conflicting results, as have prospective studies based on stored blood samples. Two large prospective studies found that persons who had used vitamin E supplements for > or = 2 y had approximately 40% lower rates of CHD. Short durations and doses of < 100 IU/d had no significant effect. The effect of dietary vitamin E was modest and nonsignificant. Adjustment for a wide array of other coronary risk factors had little effect on the findings, which were specific for vitamin E and not other supplements. The only large, randomized trial found no material reduction in CHD risk for 50 IU vitamin E/d. The epidemiologic evidence suggests that high doses of vitamin E may reduce the risk of CHD. PMID- 7495233 TI - Epidemiologic evidence of a role of carotenoids in cardiovascular disease prevention. AB - The tremendous chemical potential of the highly conjugated double bonds in carotenoids has driven research into their protective role in cardiovascular disease development. Prevention of low-density-lipoprotein oxidation and reduction of oxidative stress at the plaque formation are popular hypotheses underlying this research. Many epidemiologic studies have examined relations between beta-carotene exposure and cardiovascular disease risk. These studies used different measures to determine carotenoid exposure: semiquantitative food frequency questionnaires, carotenoid concentrations in serum taken before the onset of disease and analyzed after diagnosis, and carotenoid concentrations in adipose tissue. Although the epidemiologic evidence is consistent with a protective association between beta-carotene and cardiovascular disease, findings from the first single intervention trial conducted in a large free-living population cast doubts on the utility of beta-carotene for all high-risk populations. Beta-Carotene may only represent a marker of dietary behavior conductive to lower risk of cardiovascular disease. Research on other carotenoids is needed. PMID- 7495234 TI - Antioxidant vitamin-cardiovascular disease hypothesis is still promising, but still unproven: the need for randomized trials. AB - The hypothesis that antioxidant vitamins might decrease the risk of cardiovascular disease (CVD) is a promising area of research. At present, however, it is far from certain whether antioxidant vitamins confer protection against CVD. Evidence for the antioxidant vitamin-cardiovascular disease hypothesis has accumulated from several lines of research. Laboratory research has identified biochemical properties of antioxidant vitamins that could explain their possible role in inhibiting and delaying coronary atherosclerosis. Epidemiologic studies have provided support for the hypothesis by showing that people who consume high amounts of antioxidant vitamins through diet or supplements, or those with high concentrations of these nutrients in their blood, tend to have lower risks of CVD. In the case of the former, however, laboratory findings may not have relevance to free-living humans. Observational epidemiologic studies cannot exclude the possibility that people who consume antioxidant-rich diets or who take vitamin supplements also share other lifestyle or dietary practices that actually account for their lower disease rates. Because of these uncertainties, the only way to determine reliably whether antioxidants play any role in reducing the risk of CVD is to conduct large-scale, randomized trials of these agents, in which adequate doses of antioxidant vitamins are tested for a sufficient duration to allow for any benefits to emerge. Several large-scale trials are now ongoing in both primary and secondary prevention. The results of these trials over the next several years should provide reliable evidence for this promising, but as yet unproven, hypothesis. PMID- 7495235 TI - Vitamin E plus aspirin compared with aspirin alone in patients with transient ischemic attacks. AB - One hundred patients with transient ischemic attacks, minor strokes, or residual ischemic neurologic deficits were enrolled in a double-blind, randomized study comparing the effects of aspirin plus vitamin E [0.4 g (400 IU)/d; n = 52] with aspirin alone (325 mg; n = 48). The patients received study medication for 2 y or until they reached a termination point. Preliminary results show a significant reduction in the incidence of ischemic events in patients in the vitamin E plus aspirin group compared with patients taking only aspirin. There was no significant difference in the incidence of hemorrhagic stroke although both patients who developed it were taking vitamin E. Platelet adhesion was also measured in a randomized subgroup of both study populations by using collagen III as the adhesive surface. There was a highly significant reduction in platelet adhesiveness in patients who were taking vitamin E plus aspirin compared with those taking aspirin only. Measurement of alpha-tocopherol concentrations confirmed compliance of the patients with the medication schedule, showing a near doubling of serum concentrations of alpha-tocopherol. We concluded that the combination of vitamin E and a platelet antiaggregating agent (eg, aspirin) significantly enhances the efficacy of the preventive treatment regimen in patients with transient ischemic attacks and other ischemic cerebrovascular problems. PMID- 7495237 TI - Epidemiologic evidence for beta-carotene and cancer prevention. AB - In 1981 it was hypothesized that a high dietary intake of beta-carotene might reduce human cancer rates. Since then, several observational epidemiologic studies have addressed this topic. The results of both case-control and cohort studies show a remarkable consistency for the association of increased lung cancer risk with low amounts of dietary beta-carotene or low plasma beta-carotene concentrations. For stomach cancer, the evidence is also consistent, although the number of studies is more modest. For breast and prostate cancer, the studies indicate no consistent association of plasma or dietary beta-carotene and reduced cancer risk. For colorectal cancer, the effect will be moderate, if existent. For several other cancer sites, the numbers of cases in prospective studies are often small, implying that only strong associations can be detected. For some of these sites, results from retrospective studies are promising. The epidemiologic studies should be carefully interpreted because dietary habits may be misclassified and smoking may reduce plasma beta-carotene concentrations. Observational epidemiology cannot definitively resolve whether associations are indeed due to beta-carotene, or to other components of fruit and vegetables that are rich in beta-carotene. However, overall results are promising and several plausible cancer preventive mechanisms have been reported for beta-carotene. The ongoing human intervention studies will provide more answers regarding cancer prevention by beta-carotene but may need long follow-ups to be conclusive. PMID- 7495236 TI - Epidemiologic evidence for vitamin C and vitamin E in cancer prevention. AB - Antioxidant nutrients have been hypothesized to be protective against cancer. Vitamin C is a major circulating water-soluble antioxidant, and vitamin E is a major lipid-soluble antioxidant. Many case-control and cohort studies have related cancer risk to estimates of nutrient intake derived from food intake reports. Diets high in fruit and vegetables, and hence high in vitamin C, have been found to be associated with lower risk for cancers of the oral cavity, esophagus, stomach, colon, and lung. Diets high in added vegetable oils, and hence high in vitamin E, have been less consistently shown to be associated with cancer protection. This may be because vitamin E offers less protection against cancer or because the estimation of vitamin E intake is less accurate than is the estimation of vitamin C intake. In contrast with the findings from epidemiologic studies based on foods, observational studies of nutrients consumed in supplements and recent experimental trials provide little support for a strong protective role for vitamins C or E against cancer. If vitamins C or E are indeed protective against cancer, that protection may derive from their consumption in complex mixtures with other nutrients and with other bioactive compounds as found in the matrix provided by whole foods. PMID- 7495238 TI - Evidence from cancer intervention and biomarker studies and the development of biochemical markers. AB - Evidence from intervention trials with vitamins E and C and beta-carotene are reviewed as well as evidence from trials that have used intermediate endpoints with a special emphasis on biomarkers of cancer of the colorectum. The methodologic issues that require resolution before a second generation of clinical trials are launched to assess the efficacy of these antioxidants in the prevention of cancer are identified. Specific concerns regarding the validation of pathologic biomarkers of cancer and biochemical markers of mechanism of action for the antioxidants are discussed. Cellular proliferation indexes in the colon are used as an example of pathologic biomarkers for cancer and the measurement of plasma and tissue malondialdehyde concentrations is used as an example of problems with the development of biochemical markers of oxidative stress that can be used in prevention trials. The use of DNA oxidation products as promising biomarkers is also discussed. PMID- 7495239 TI - Antioxidants in oral cancer prevention. AB - I present evidence in support of a chemopreventive role for the so-called antioxidant nutrients, beta-carotene and vitamin E, against oral cavity cancer. This evidence is from laboratory studies, animal model systems, epidemiologic surveys, intervention trials involving reversal of premalignant changes, and prevention of malignancies in particularly high-risk subjects. Because agents proposed for disease prevention are meant to be used widely without close medical supervision, almost any toxicity is unacceptable. beta-Carotene and vitamin E fulfill this criterion for a suitable chemopreventive agent. In several epidemiologic studies, low intakes of vitamin E, carotenoids, or both have been associated with a higher cancer risk. Smoking, a major risk factor, results in lower beta-carotene concentrations in plasma and oral mucosal cells. In several laboratory and animal model systems, beta-carotene and other antioxidant nutrients are inhibitors of oral cavity carcinogenesis. beta-Carotene and vitamin E can produce clinical regression of oral leukoplakia, a premalignant lesion for oral cancer. The design and limitations of such studies in oral leukoplakia are discussed. Cancer incidence reduction trials in high-risk groups have targeted prevention of second malignancies in patients cured of a primary oral cancer. These trials are in progress. The data thus far are supportive of a significant preventive role for these nutrients in oral cancer. PMID- 7495240 TI - Prevention of cervical intraepithelial neoplasia and cervical cancer. AB - We review the current status of prevention trials in the management of cervical intraepithelial neoplasia (CIN) and cervical cancer. Two large, randomized controlled trials have shown that folic acid is inactive in reversing low to moderate grade CIN. A large randomized trial of locally applied beta-trans retinoic acid showed that the agent was effective in reversing moderate but not severe CIN. Results from a pilot trial involving 30 patients with CIN I (mild dysplasia) and CIN II (moderate dysplasia) indicate that beta-carotene can suppress CIN; a large ongoing randomized trial will answer the question more definitively. PMID- 7495241 TI - Effect of nutrition intervention on intermediate endpoints in esophageal and gastric carcinogenesis. AB - A nutrition intervention trial involving > 3000 participants was conducted in Linxian, China, where the esophageal and stomach cancer mortality rates are among the highest in the world and suspicion exists that chronic deficiencies of multiple nutrients are etiologically involved. The trial was randomized, double blind, and placebo-controlled and tested the effect of multivitamin and multimineral supplements in reducing cancer incidence and mortality in adults with cytologically detected esophageal dysplasia. Endoscopic and cytologic examinations of samples of trial participants during the intervention allowed evaluation of intermediate endpoints in esophageal and gastric carcinogenesis, including asymptomatic histologic precancerous lesions and early invasive cancer, epithelial proliferation, and cytologic abnormalities. Results from these ancillary studies suggest that multivitamin and multimineral supplementation may decrease proliferation and enhance cytologic reversion to nondysplasia. PMID- 7495242 TI - The Linxian trials: mortality rates by vitamin-mineral intervention group. AB - Two randomized nutrition intervention trials were conducted in Linxian, an area of north central China with some of the world's highest rates of esophageal and stomach cancer and a population with a chronically low intake of several nutrients. One trial used a factorial design that allowed us to assess the effects in nearly 30,000 participants of daily supplementation with four nutrient combinations: retinol and zinc; riboflavin and niacin; vitamin C and molybdenum; and beta-carotene, alpha-tocopherol, and selenium. The second trial provided daily multiple vitamin-mineral supplementation or placebo in 3318 persons with esophageal dysplasia, a precursor to esophageal cancer. After supplements were given for 5.25 y in the general population trial, small but significant reductions in total [relative risk (RR) = 0.91] and cancer (RR = 0.87) mortality were observed in subjects receiving beta-carotene, alpha-tocopherol, and selenium but not the other nutrients. The reductions were greater in women than men, and in those under compared with over the age of 55; however, differences by sex or age were not significant. After multiple vitamin and mineral supplements were given for 6 y in the smaller dysplasia trial, reductions in total (RR = 0.93) and cancer (RR = 0.96) mortality were observed but these were not significant. The largest reductions were for cerebrovascular disease mortality, but the effects differed by sex: a significant reduction was observed in men (RR = 0.45) but not women (RR = 0.90). Restoring adequate intake of certain nutrients may help to lower the risk of cancer and other diseases in this high-risk population. PMID- 7495243 TI - Effects of alpha-tocopherol and beta-carotene supplements on cancer incidence in the Alpha-Tocopherol Beta-Carotene Cancer Prevention Study. AB - The Alpha-Tocopherol Beta-Carotene (ATBC) Cancer Prevention Study was a placebo controlled, randomized intervention trial testing the hypothesis that beta carotene and alpha-tocopherol (vitamin E) supplements prevent lung and other cancers. The study is predicated on a substantial body of evidence supporting a role in cancer prevention for these micronutrients. Based on the 2 x 2 factorial study design, 29,133 eligible male cigarette smokers aged 50-69 y were randomly assigned to receive beta-carotene (20 mg), alpha-tocopherol (50 mg), beta carotene and alpha-tocopherol, or placebo daily for 5-8 y. Capsule compliance was high (median = 99%). beta-Carotene treatment did not result in a decrease in cancer at any of the major sites but rather in an increase at several sites, most notably lung, prostate, and stomach (number of cases 474 compared with 402, 138 compared with 112, and 70 compared with 56, respectively). The vitamin E group had fewer incident cancers of the prostate and colorectum compared with the group not receiving vitamin E (number of cases 99 compared with 151 and 68 compared with 81, respectively), but more cancers of the stomach (70 compared with 56). In contrast to these intervention-based findings for beta-carotene and vitamin E supplements, we observed lower lung cancer rates in men with higher amounts of both serum and dietary beta-carotene and vitamin E at baseline. PMID- 7495244 TI - Beta-Carotene, vitamin A, and lung cancer chemoprevention: results of an intermediate endpoint study. AB - A randomized, placebo-controlled clinical trial of beta-carotene and retinol was conducted with 755 former asbestos workers as study subjects. The targeted endpoint for the intervention study was a reduction in the incidence and prevalence of sputum atypia. The dosage of 50 mg beta-carotene/d and 25,000 IU retinol/d on alternate days resulted significant increases in serum concentrations of both agents with no clinically significant toxicity. Skin yellowing was observed in approximately 35% of patients and may have contributed adversely to protocol adherence. Baseline analysis revealed that smoking and drinking were associated with lower concentrations of serum beta-carotene, even after dietary carotene intake was adjusted for (P < 0.0001). Baseline concentrations of retinol were apparently lowered by smoking (P < 0.002) and increased by drinking (P < 0.0001). Drinking and smoking also were significantly related to lower beta-carotene concentrations after supplementation (P < 0.001). No significant reduction in sputum atypia was observed after treatment. PMID- 7495245 TI - Relations among aging, antioxidant status, and cataract. AB - Light and oxygen are necessary for the function of the eye. However, when present in excess or in uncontrolled circumstances, they appear to be related, probably causally, to the development of cataract. Compromises of function of the lens and retina with aging are exacerbated by depleted or diminished primary antioxidant reserves, antioxidant enzyme capabilities, and diminished secondary defenses such as proteases. Smoking appears to provide an additional oxidative challenge associated with depletion of antioxidants as well as with enhanced risk for cataract formation. Poor education and lower socioeconomic status are associated with poorer nutriture and are also significantly related to increased risk for these debilities. Optimizing nutriture, including diets rich in fruit and vegetables, may provide the least costly and most practicable means to delay cataract. PMID- 7495246 TI - Evidence for protection against age-related macular degeneration by carotenoids and antioxidant vitamins. AB - Epidemiologic data indicate that individuals with low plasma concentrations of carotenoids and antioxidant vitamins and those who smoke cigarettes are at increased risk for age-related macular degeneration (AMD). Laboratory data show that carotenoids and antioxidant vitamins help to protect the retina from oxidative damage initiated in part by absorption of light. Primate retinas accumulate two carotenoids, lutein and zeaxanthin, as the macular pigment, which is most dense at the center of the fovea and declines rapidly in more peripheral regions. The retina also distributes alpha-tocopherol (vitamin E) in a nonuniform spatial pattern. The region of monkey retinas where carotenoids and vitamin E are both low corresponds with a locus where early signs of AMD often appear in humans. The combination of evidence suggests that carotenoids and antioxidant vitamins may help to retard some of the destructive processes in the retina and the retinal pigment epithelium that lead to age-related degeneration of the macula. PMID- 7495247 TI - Antioxidants and immune response in aged persons: overview of present evidence. AB - The oxidant-antioxidant balance is an important determinant of immune cell function, including maintaining the integrity and functionality of membrane lipids, cellular proteins, and nucleic acids and controlling signal transduction and gene expression in immune cells. Optimal amounts of antioxidants are needed for maintenance of the immune response across all age groups. This need might be more critical, however, in aged persons. Age-associated dysregulation of immune response, particularly of T cell-mediated function, is well documented. The well known age-related increase in free radical formation and lipid peroxidation contributes, at least in part, to this phenomenon. We summarize animal and human studies undertaken by ourselves as well as other investigators on the effects of antioxidants, vitamin E, beta-carotene, and glutathione on the immune response of aged persons. The underlying mechanisms for the antioxidant nutrients' effects as well as their health implications for aged persons are discussed. PMID- 7495248 TI - Possible immunologic involvement of antioxidants in cancer prevention. AB - The people of Linxian County, China have one of the world's highest rates of esophageal cancer. Two intervention trials were conducted to determine whether supplementation with specific vitamins and minerals could lower mortality from or incidence of cancer in this population and whether supplementation with multiple vitamins and minerals would reduce esophageal and gastric cardia cancer in persons with esophageal dysplasia. About 30,000 general population (GP) subjects in the GP trial were randomly assigned to one of eight intervention groups according to a one-half replicate of a 2(4) factorial experimental design and were supplemented for 5.25 y with four combinations of micronutrients at doses from one to two times the US recommended dietary allowance (RDA). About 3000 subjects in whom dysplasia was diagnosed in the dysplasia trial were randomly assigned to groups receiving daily supplementation with 14 vitamins and 12 minerals at two to three times the US RDA or placebo for 6 y. Results of the dysplasia trial indicate that in individuals with esophageal dysplasia, micronutrient supplementation had little effect on T lymphocyte responses. In contrast, male participants in the GP trial who were supplemented with beta carotene, vitamin E, and selenium showed significantly (P < 0.05) higher mitogenic responsiveness of T lymphocytes in vitro than those not receiving these micronutrients. PMID- 7495249 TI - Dietary and pharmacologic regimens to reduce lipid peroxidation in non-insulin dependent diabetes mellitus. AB - Persons with diabetes are at increased risk for developing coronary heart disease (CHD): although the standard risk factors are also applicable, there are other major nontraditional risk factors for these persons. For example, oxidation of lipoproteins may play an important role in the development of atherosclerosis. Another risk factor is the presence of small, dense low-density lipoprotein (LDL), which may have enhanced atherogenicity because it enters the arterial wall more readily and is more easily oxidized. Both of these factors may be particularly important in persons with non-insulin-dependent diabetes mellitus (NIDDM), because these persons may have greater rates of lipid oxidation in vivo than do nondiabetic persons and also have an increased prevalence of small, dense LDL. I describe potential dietary and pharmacologic regimens in patients with NIDDM to decrease in vivo lipid peroxidation, and the susceptibility of LDL and dense LDL subfractions to oxidative modification. In nearly all NIDDM patients, reducing dietary saturated fat helps lower plasma cholesterol and reduces the risk for CHD. There is, however, some controversy as to whether dietary saturated fat should be replaced by carbohydrates or by monounsaturated fatty acids (MUFAs) in NIDDM. Recent studies showed reduced susceptibility to oxidation of LDL in subjects consuming MUFA-enriched diets, thus adding another dimension to the ongoing debate over the most appropriate diet for NIDDM patients. Additionally, supplemental antioxidants such as probucol and vitamin E alone or in combination with MUFA-enriched diets have shown promise in protecting LDL from oxidation when given to nondiabetic populations. I also present recent results from an antioxidant trial in NIDDM subjects. PMID- 7495250 TI - Dietary antioxidants and cigarette smoke-induced biomolecular damage: a complex interaction. AB - Epidemiologic evidence suggests that cigarette smoking is a major risk factor for chronic obstructive pulmonary diseases such as chronic bronchitis and emphysema, for carcinogenesis, and for cardiovascular disease. However, the precise mechanisms of these effects are incompletely understood. The gas phase of cigarette smoke contains abundant free radicals including nitric oxide. Hence, cigarette smoke may induce some of its damaging effects by free radical mechanisms. We report that exposure of plasma, a model for respiratory tract lining fluids, to gas-phase cigarette smoke causes depletion of antioxidants, including ascorbate, urate, ubiquinol-10, and alpha-tocopherol, and a variety of carotenoids, including beta-carotene. Gas-phase cigarette smoke induced some lipid peroxidation, as measured by cholesteryl linoleate hydroperoxide (18:2OOH) formation. Ascorbate was effective in preventing 18:2OOH formation. In contrast to the low concentrations of lipid hydroperoxides measured (< 1 mumol/L), protein carbonyl formation, a measure of protein modification, increased by approximately 400 mumol/L after nine puffs of cigarette smoke. Reduced glutathione inhibited protein carbonyl formation, whereas other plasma antioxidants, including ascorbate, were ineffective. alpha, beta-Unsaturated aldehydes (acrolein and crotonaldehyde) in cigarette smoke may react with protein -SH and -NH2 groups by a Michael addition reaction that results in a protein-bound aldehyde functional group. Gas-phase cigarette smoke is capable of converting tyrosine to 3 nitrotyrosine and dityrosine, indicating free radical mechanisms of protein damage by nitrogen oxides. Aldehydes and nitrogen oxides in cigarette smoke may be significant contributors to biomolecular damage, and endogenous antioxidants can attenuate some of these adverse effects. PMID- 7495251 TI - Vitamin E: beyond antioxidant function. AB - Vitamin E, a potent peroxyl radical scavenger, is a chain-breaking antioxidant that prevents the propagation of free radical damage in biological membranes. We consider the evidence for potential sites in cellular metabolism and signal transduction where vitamin E may have a structure-specific role in addition to its antioxidant function. The roles of tocopherol-binding proteins in cellular trafficking of vitamin E, especially the incorporation of RRR-alpha-tocopherol into nascent lipoproteins, and the delivery of RRR-alpha-tocopherol to the nucleus are considered. We discuss the functions of vitamin E both in the inhibition and potentiation of arachidonic acid metabolism. The interactions of vitamin E during cell proliferation and differentiation are also evaluated. These functions of vitamin E raise new questions and represent new and exciting areas for research in cell regulation with physiologic implications. PMID- 7495252 TI - Safety of antioxidant vitamins and beta-carotene. AB - Epidemiologic evidence links high antioxidant status with low risk of degenerative disease. Optimal intakes of antioxidants may not be achievable by diet alone; supplements may be taken, particularly in subgroups of the population at high risk. It is thus necessary to ensure that antioxidant supplements are safe and free from side effects. The toxicity of vitamin E is low; no mutagenic, teratogenic, or carcinogenic effects are known and in double-blind studies in which large amounts of vitamin E were used in humans, no side effects occurred. High concentrations are contraindicated in subjects with vitamin K-associated blood coagulation disorders, and the toxicity in normal subjects ingesting large amounts of vitamin E over long periods requires additional investigation. Toxicity of beta-carotene also is low. Evidence from human toxicity trials is not available but there is much circumstantial evidence that 15-50 mg/d is without side effects except for hypercarotenemia in some subjects at high intakes. The findings of more lung cancer in subjects who smoked and who were given 20 mg beta carotene/d than in those given a placebo could be influenced by the cancer being well advanced before beta-carotene administration. Massive anecdotal evidence exists that vitamin C (at > or = 1 g/d) is safe. Exhaustive literature searches have failed to reveal a controlled study of vitamin C toxicity in human subjects. Anxiety exists about oxalate stone formation, uricosuria, vitamin B-12 destruction, mutagenicity, and iron overload, but the consensus is that adverse effects do not occur in healthy subjects ingesting large amounts of vitamin C. PMID- 7495253 TI - Are clinical trials really the answer? AB - It has been asserted that clinical trials hold the answer to questions about the role of nutrients in preventing chronic diseases. This is not the case. Clinical trials give us rigorous answers to restricted questions. Rarely can more than one or two substances be tested, usually at a single dose. Subjects usually have to be persons with precancerous conditions or an extremely high risk of the disease in question. Rarely can any diseases other than the most common ones be studied. Most important, clinical trials test the efficacy of an agent that is administered for a limited time, beginning fairly late in life. Few trials will tell us anything about whether dietary amounts of nutrients might contribute to prevention of long-term chronic diseases. They also tell us nothing about whether agents at high doses might reduce disease risk if taken throughout the lifetime. Furthermore, they tell us nothing about other antioxidants, other combinations, or other doses. Clinical trials were developed for therapeutic situations to determine which treatment was better for curing a specific disease. However, the questions about prevention that are of interest may involve persons with no unusual risk of disease, lifetimes of exposure, enormously complex interactions among nutrients, and the effects of these nutrients on hundreds of often uncommon disease conditions. Clinical trials simply cannot answer these questions. Only a solid examination of the laboratory and epidemiologic evidence can approximate the answers to most of the questions of interest. PMID- 7495254 TI - Considerations of the scientific substantiation for antioxidant vitamins and beta carotene in disease prevention. AB - Advances in our knowledge about the role vitamin C, vitamin E, and beta-carotene may play in reducing the risk of chronic diseases have been derived concomitantly from several different research approaches, each with its own advantages and limitations. Evaluation of the evidence for potential health benefits of antioxidant nutrients should include the entire spectrum of available scientific evidence--from cell biology, animal studies, clinical trials, and epidemiologic surveys--and consider the quality, strength, consistency, and biological plausibility of this evidence. However, difficult questions arise when considering how strong the correlation should be between the totality of evidence and the conclusions drawn for making individual and public health recommendations. Many of these questions address the need for additional research not only directed to the efficacy and safety of these nutrients but to their bioavailability, interactions with one another and other dietary factors, mechanisms of action, and methods to assess their functional status. PMID- 7495255 TI - Novel urinary metabolite of alpha-tocopherol, 2,5,7,8-tetramethyl-2(2' carboxyethyl)-6-hydroxychroman, as an indicator of an adequate vitamin E supply? AB - Previously, the metabolism of alpha-tocopherol was considered to involve the opening of the chroman structure because of its oxidation to tocopherylquinone. In contrast, we describe here 2,5,7,8-tetramethyl-2(2'-carboxyethyl)-6 hydroxychroman (alpha-CEHC) as the major urinary metabolite of alpha-tocopherol that appears in human urine after vitamin E supplementation. It is formed directly from alpha-tocopherol without previous oxidative splitting of the chroman ring. The correlation of alpha-tocopherol intake, plasma alpha-tocopherol concentrations, and urinary excretion of alpha-CEHC in human volunteers supplemented with RRR-alpha-tocopherol dosages ranging from 0 to 800 mg/d was examined. HPLC and gas chromatography-mass spectroscopy analysis revealed that alpha-CEHC was only excreted when a plasma threshold of 7-9 mumol alpha tocopherol/g total lipid was exceeded. This concentration was obtained by a daily intake of approximately 50-150 mg alpha-tocopherol. We suggest that alpha-CEHC excretion indicates a saturated binding capacity of vitamin E in the plasma and thus may be considered to be a marker of optimum vitamin E intake. PMID- 7495257 TI - Oleoresin capsicum (pepper) spray and "in-custody deaths". AB - Increasing use of oleoresin capsicum (OC) spray devices (i.e., pepper spray, pepper mace, OC, capsaicin) by law enforcement agencies as a means of sublethal force to control suspects has brought into question whether exposure to this noxious irritant (capsaicin) can cause or contribute to unexpected in-custody deaths. Capsaicin stimulates nociceptors in exposed mucous membranes to produce intense pain, particularly involving the conjunctiva, and generates systemic physiologic and behavioral responses consonant with such extreme discomfort. We describe two cases of in-custody death, both associated temporally with the use of pepper spray, to illustrate salient investigative considerations. As with any other in-custody death, a thorough autopsy and toxicologic analysis, coupled with evaluation of the premortem chain of events, postexposure symptomatology, and the extent of natural disease processes, will help to reveal the role of oleoresin capsicum spray as unrelated, contributory, or causative. PMID- 7495256 TI - Growth retardation in human cervical dysplasia-derived cell lines by beta carotene through down-regulation of epidermal growth factor receptor. AB - We used newly established cervical dysplasia-derived cell lines to elucidate a molecular mechanism of the preventive action of beta-carotene in cervical multi step carcinogenesis. Liposomal beta-carotene was added to the culture medium for human cervical dysplasia cell lines, CICCN-2 from cervical intraepithelial neoplasia grade I (CIN I), CICCN-3 from CIN II, and CICCN-4 from CIN III, and human cervical carcinoma-derived cell lines such as CICCN-6, CICCN-18, and HeLa cells. beta-Carotene (10 mumol/L) induced significant growth retardation in three cervical dysplasia cell lines but not in three cervical carcinoma-derived cell lines. Binding activities of epidermal growth factor (EGF) and cellular amounts of either messenger RNA for EGF receptor gene or EGF receptor protein were all highest in CICCN-4 cells. Cell surface binding, as well as internalization, of 125I-labeled EGF was rapidly reduced after beta-carotene treatment in dysplasia cell lines and 170-kD protein bands of EGF receptor disappeared from protein immunoblots at day 3 of the treatment. Cellular amounts of EGF receptor messenger RNA remained constant until day 3 of the treatment and were substantially reduced after day 7. Chromatin condensations, morphologic evidence for apoptotic cell death, were observed at day 1 by staining. From these results, we contend that prevention of cervical carcinogenesis by beta-carotene is due to induction of apoptosis in cervical dysplastic cells, which are premalignant cells in cervical multi-step carcinogenesis, via down-regulation of EGF receptor protein. PMID- 7495258 TI - Pathologic changes of the cardiac conduction tissue in sudden cardiac death. AB - We report the pathologic changes of the cardiac conduction tissue (CS) in 33 cases of sudden cardiac death. The cases were divided into two groups: those with severe atherosclerotic coronary lesion (> 75% cross-sectional area stenosis of at least one coronary artery), 15 cases, and those without significant coronary disease, 18 cases. In the first group, we observed major changes of the CS in eight cases. The changes consisted of abnormalities in His bundle position (branching bundle in pars membranacea or left-sided bundle), atrioventricular (AV) node fibrosis, and interatrial lipoma. Other age-related minor abnormalities such as mild fibrosis, fatty change, or fibrointimal hyperplasia of CS arteries were observed in nine cases. In the second group, major lesions of the CS were recognized in 13 cases: abnormalities in bundle branch position in eight cases (left-sided, inside the membranous septum), downward displacement of the tricuspid valve with AV node elongation in three cases, and a single case with one of the following: His compression by hypertrophic cardiomyopathy, Mahaim fibers, focal ischemia of bundle, AV node division, AV node dissection by edema, intramural left bundle branch, or tumor of the AV node. In the study of the pathogenesis of sudden death, the conduction system lesions always have to be considered, even in cases with severe coronary disease. PMID- 7495259 TI - Immunohistochemical parameters for the age estimation of human skin wounds. A review. AB - The immunohistochemical detection of stringlike ramifying structures positive for fibronectin in wounds aged at least a few minutes provides valuable information on the vitality of skin lesions even before the appearance of neutrophilic granulocytes. Positive reactions for tenascin or collagen type III indicate postinfliction intervals of at least 2-3 days, whereas vital reactions for collagen type V or VI occur earliest 3 days after wounding. Collagen type I appears as spotlike fibroblast-associated reaction products in injuries aged 4 days or more, while typical stringlike ramifying fibers indicate a postinfliction interval of at least 5-6 days. Fibroblasts positively staining for laminin or heparan sulfate proteoglycan can be detected in wounds with a survival time of approximately 1.5 days or more and collagen type-IV-positive cells occur earliest 4 days after wounding, followed by alpha-smooth muscle actin expressing fibroblasts after 5 days or more. Basement membrane fragments positive for laminin, heparan sulfate proteoglycan, or collagen type IV or VII indicate a wound age of at least 4 days, and a complete restitution of the epidermal basement membrane in (surgical) wounds can be observed earliest 8 days after wound infliction. Positively reacting basement membrane fragments are absent up to a wound age of < 13 days, and the evidence of an incompletely rebuilt basement membrane indicates a postinfliction interval of less than approximately 22 days. A continuous staining of the basal cells of the newly formed epidermis for cytokeratin 5 occurs earliest 13 days after wounding, and the absence of a complete staining provides information on a wound age of < 24 days. PMID- 7495261 TI - Can baby organs be donated in all forensic cases? Proposed guidelines for organ donation from infants under medical examiner jurisdiction. AB - Donated organs from babies are desperately needed for transplantation into sick and dying children. Categories of infant death subject to medicolegal death investigation are proposed and discussed, with a rationale for decision making by medical examiners and their colleagues. Appropriate caution is warranted in some instances with investigation for possible criminal intent emphasized. Mutual cooperation with all parties must be fostered to assure organ donation from infants whenever possible. PMID- 7495260 TI - Creatine kinase BB and neuron-specific enolase in cerebrospinal fluid in the diagnosis of brain insult. AB - Cerebrospinal fluid (CSF) markers provide useful information about the extent of brain damage. These biochemical indices may also be used when postmortem histopathological examination does not confirm antemortem brain insult. Seven biochemical parameters--creatine kinase (CK), creatine kinase BB isoenzyme (CK BB), lactate dehydrogenase (LDH), gamma-glutamyltransferase, aldolase, leucine aminopeptidase (LAP), and neuron-specific enolase (NSE)--were analyzed in CSF from 82 cadavers. Case studies were categorized into one of four diagnostic groups. There were 15 cases of head trauma, 23 of hypoxia (hangings, carbon monoxide, and drug poisonings), 23 sudden cardiac death, and 21 miscellaneous cases. The degree of craniocerebral trauma was graded. In CSF there was a statistically significant correlation between the severity of craniocerebral trauma and levels of CK, CK-BB, aldolase, LDH, and LAP. CSF CK-BB [median U/L (range)] for the groupings of head trauma, hypoxia, sudden cardiac death, and miscellaneous were, respectively, 873 (1-12,100), 26 (2-2,780), 16 (1-42), and 18 (0-2,780). Corresponding CSF CK levels were 9,370 (28-67,842), 101 (18-36,840), 180 (10-29,622), and 264 (17-26,556). There were no statistical significant differences among the NSE concentrations in the four diagnostic groups. The testing of biochemical markers could be a reliable indicator of the degree of brain insult in support of morphological studies. PMID- 7495262 TI - Estimating blood alcohol from urinary alcohol at autopsy. AB - Urine alcohol concentration (UAC) and blood alcohol concentration (BAC) measured by gas chromatography were available from 435 medicolegal autopsies. Simple linear regression with BAC as outcome variable and UAC as predictor variable (range, 3-587 mg%) gave the regression equation BAC = -5.6 + 0.811UAC with 95% prediction interval +/- 0.026 square root of [9465804 + (UAC-213.3)2] and 99% prediction interval +/- 0.034 square root of [9465804 + (UAC-213.3)2]. The standard error of the slope was 0.013 and the 95% confidence interval for the slope 0.785-0.837. In practice, a BAC of 80 mg% is predicted with 95% certainty by a UAC of 204 mg% and similarly a BAC of 150% by a UAC of 291 mg%. The prediction interval is too wide to be helpful in the assessment of an individual case fatality. The UAC is useful in corroborating but not in predicting BAC. PMID- 7495264 TI - Gas flames, closed spaces, and hypoxia. AB - The investigation of the deaths of an elderly couple found in a small hotel room that was tightly sealed and heated by an open flame gas heater is presented. Autopsy and toxicologic studies did not reveal a definitive cause of death. Subsequent research and an experiment to duplicate the conditions support the contention that the cause of death was hypoxia due to oxygen depletion by the gas flame. PMID- 7495263 TI - "There's death in the cup, so beware!". AB - The proper investigation of deaths due to poisoning and the determination of the manner of death often depend upon knowledge and investigative skills that may have been allowed to deteriorate with lack of use. These case reports illustrate a wide variety of substances that people can ingest. This collection of case reports includes the ingestion of two insecticides and three different organic chemicals. A concise review of literature is provided, along with symptoms exhibited by those who ingest these poisons, and methodologies recommended for their determination in this series of cases. PMID- 7495265 TI - The lethal paraphiliac syndrome. Accidental autoerotic deaths in Denmark 1933 1990. AB - A new definition of accidental autoerotic death (AAD) is proposed. A death is an AAD if it is solitary, accidental, and caused by a lethal paraphilia. On the basis of a series of 46 AADs, all occurring among men in the period 1933-1990 in Denmark, the definition cannot be rejected. A paraphilia is regarded as lethal if it is inherently life-threatening. The results of this study have been related to previous reports of similar autoerotic deaths in the literature. It is suggested that the present distinction between asphyxial AAD as typical and nonasphyxial ADD as atypical be replaced with lethal paraphilia with accompanying nonlethal paraphilia or props as typical AAD and lethal paraphilia with no accompanying nonlethal paraphilia or props as atypical AAD. PMID- 7495266 TI - Estimation of age from the teeth of unidentified corpses using the amino acid racemization method with reference to actual cases. AB - We report the results of a study for estimating the age of unidentified corpses from teeth using the amino acid racemization method. By calculating the ratio of D-aspartic acid to L-aspartic acid (referred to as the D/L ratio), we studied samples from 24 corpses of known identity. The results showed that the approach yielded satisfactory age estimates with an accuracy of +/- 3 years. Detailed examination of these results indicated that powdered dentin produced more favorable results than fragmented dentin. No differences in the estimated age related to cause of death were noted. However, better results were obtained with a larger number of control teeth and a higher correlation coefficient between the D/L ratio and the actual age of the control teeth. In terms of age and estimation accuracy, corpses of individuals > 40 years of age generally gave more satisfactory results than those of younger individuals. It was also reconfirmed that preparation of control teeth is required each time an age estimate is performed. PMID- 7495267 TI - Plasma cell granuloma of the larynx as a cause of sudden asphyxial death. AB - A unique case of plasma cell granuloma of the larynx associated with sudden death in a young HIV-positive woman is described. The plasma cell infiltration fulfilled the criteria for plasma cell granuloma, showing characteristic histologic and immunohistochemical features (polyclonality). A review of the most recent literature is provided. PMID- 7495268 TI - "Tentative" injuries in a homicide. AB - A homicide caused by blunt trauma to the neck and head is reported. On the left side of the victim's neck were multiple superficial cuts that ran parallel to deeper ones that appeared to be the typical pattern of tentative or hesitation injuries. Further information on the perpetration of the crime could not be obtained from the offender, but it must be assumed that the cuts were made after the victim had already been knocked unconscious by blows or kicks. The considerable blood alcohol concentration of the perpetrator and the rather reduced sharpness of the serrated kitchen knife used probably contributed to the development of these injuries. PMID- 7495269 TI - Organomegaly in chronic alcoholics. AB - We compared the weights of the brain, heart, spleen, lungs, liver, and kidneys, uncorrected and corrected for body surface area (BSA) and body mass index (BMI), of 50 alcoholics with 50 forensic controls matched for sex, age, and body weight. With the exception of combined renal weight corrected for BSA and BMI, no significant differences were found in organ weights between the two groups. Glomerular diameter corrected for BSA was significantly greater in alcoholics than controls. It appears that nephromegaly and glomerulomegaly are both population markers for alcoholism. In alcoholics, there was no correlation between the microscopic degree of steatosis and glomerulomegaly. In controls, renal glomerular diameter was greater with increasing severity of hepatic steatosis, probably as a reflection of obesity. PMID- 7495270 TI - Pleural effusion in drowning. AB - The factors that determine the amount of pleural effusion in cases of drowning have not been well established. In a series of cases of drowning from the years 1987-1991, several parameters have been registered to determine whether any correlations could be found between these and the amount of pleural fluid in the drowning victims. A statistically significant difference was found between the cases with and those without increased pleural effusion, indicating that the time spent in water is correlated to the production of pleural fluid. A significant difference was also found between saltwater and freshwater drownings, indicating that more pleural fluid is produced in saltwater than in freshwater drownings. PMID- 7495271 TI - Decreasing the organ donor shortage. PMID- 7495272 TI - Decreasing the organ donor shortage. PMID- 7495273 TI - Spreading knowledge? PMID- 7495274 TI - Short and sweet: looking for sugar. PMID- 7495275 TI - Pathology and emerging infections--quo vadimus? AB - There have been dramatic changes in the occurrence of infectious diseases throughout the world in the previous two decades. The emergence of new microbial agents, and the reemergence of infections previously believed to be controlled, threatens the health of all populations. The emergence of these infectious diseases has occurred during a period of breakdown in the capabilities of the public health surveillance systems, prevention programs, and disease control efforts. Expertise in pathology is critical to provide a strong national control program for emerging and reemerging infectious diseases. Despite the many significant achievements made by pathologists in improving understanding of the pathogenesis and diagnosis of infectious diseases, the field of pathology remains largely oriented toward neoplastic diseases, and has not yet identified infectious disease diagnosis as an important component of anatomic pathology training and research, even in the face of the current threats posed by microbial agents. In addition, there is currently a dearth of infectious disease pathologists in the United States and elsewhere in the world, and the use of the autopsy, a prime pathological tool for diagnosis of emerging infections, is on the wane. The most serious problem is that no formal training program for infectious disease pathology currently exists in the United States or elsewhere in the world. As a consequence of this lack of training opportunities, there is a severe deficiency of young, well-educated pathologists with infectious disease expertise. This article explores the historical linkages between the disciplines of infectious diseases and pathology, and suggests that infectious disease pathology as a subspecialty be strengthened and training programs be initiated. PMID- 7495276 TI - Interleukin-12 and interferon-gamma. Do they always go together? PMID- 7495277 TI - RET proto-oncogene mutations and rearrangements in endocrine diseases. PMID- 7495278 TI - Membrane transport proteins associated with drug resistance expressed in human melanoma. AB - Melanoma cells often display a multidrug-resistant phenotype, but the mechanisms involved are largely unknown. We have studied here the recently identified transport-associated proteins, MRP and LRP, and the well-known drug resistance marker P-glycoprotein using a panel of 16 human melanoma cell lines and 71 benign and malignant melanocytic tissue samples. By flow cytometry and immunohistochemistry, expression of P-glycoprotein was not detectable on the protein level in the 10 cell lines analyzed, although by reverse transcriptase polymerase chain reaction, MDR-1 gene expression was demonstrated in 2 of 10 cell lines. In addition, immunohistology revealed P-glycoprotein expression in only 1 of 71 melanocytic lesions. In contrast, MRP was detected in a subset of melanoma cell lines by reverse transcriptase polymerase chain reaction and immunohistology (4 of 10). LRP expression was observed in 8 of 10 melanoma cell lines by immunochemistry and in 10 of 10 by reverse transcriptase polymerase chain reaction. Furthermore, MRP was detected immunohistologically in almost 50% of primary and metastatic melanoma specimens, although no significant differences were found between metastases taken before or after chemotherapy. Expression of LRP was detected in a subset of nevi with nevus cells exhibiting up to 25% positive LRP reactivity. In 13 of 21 primary melanomas and 23 of 37 metastases, more than 25% of tumor cells were stained by the LRP-56 monoclonal antibody. Particularly in the group of metastases with more than 50% of LRP-positive cells, 7 of 11 of the metastases had been previously exposed to chemotherapeutic drugs. Although the expression of membrane transport proteins may explain only the chemoresistance toward lipophilic, natural compounds and not resistance against alkylating agents, the lack of P-glycoprotein expression after chemotherapeutic treatment and the significant expression of MRP and LRP in melanoma cells provide first insights into the drug-resistant phenotype in melanoma. Additional studies analyzing the role of MRP and LRP in chemoresistance of melanoma are warranted. PMID- 7495279 TI - Ring chromosomes in dermatofibrosarcoma protuberans are composed of interspersed sequences from chromosomes 17 and 22. AB - Ring chromosomes are found in most dermatofibrosarcoma protuberans (DFSPs), and recent reports demonstrate that portions of the DFSP ring chromosomes derive from chromosome 17. In this study we characterized ring chromosomes in three DFSPs using a combined approach of karyotyping, chromosome painting, and comparative genomic hybridization. Chromosome painting demonstrated that the ring chromosomes in each DFSP were composed of discontinuous, interwoven sequences from chromosomes 17 and 22. Amplification of chromosomes 17 and 22 sequences was confirmed in each of these cases by comparative genomic hybridization, and over representation of chromosomes 17 and 22 sequences was also demonstrated by comparative genomic hybridization in 1 of 2 cytogenetically unremarkable DFSPs. We conclude that amplification of chromosomes 17 and 22 sequences, in ring form, is a characteristic aberration in DFSP. PMID- 7495280 TI - Detection of mRNAs for urokinase-type plasminogen activator, its receptor, and type 1 inhibitor in giant cell tumors of bone with in situ hybridization. AB - Although giant cell tumor of bone (GCT) is generally considered to be an uncommon benign neoplasm, it can pursue an aggressive course with local recurrence and metastasis. Attempts to predict the biological behavior of GCT with histopathological parameters, however, have not been successful. The urokinase type plasminogen activation system has been implicated in tumor invasion and metastasis and abnormalities of the components of this system have been found in several malignancies. In this study we postulated that the urokinase-type plasminogen activation system associated with bone destruction and local invasion is present in GCT. We therefore evaluated the mRNA levels for urokinase-type plasminogen activator (u-PA), urokinase-type plasminogen activator receptor (u PAR), and plasminogen activator inhibitor type 1 (PAI-1) by using Northern blot analysis and in situ hybridization in four cases of GCT and spindle-shaped mononuclear cells at the 35th passage from a GCT. Our results showed that giant cell tumors of bone contained variable levels of u-PA, u-PAR, and PAI-1 mRNA, respectively, 2.3, 1.4, and 3.2 kb in size. In situ hybridization showed that u PA, u-PAR, and PAI-1 mRNA were expressed in both the mononuclear cells and the osteoclast-like giant cells; the signal for u-PA mRNA in the spindle-shaped mononuclear cells was more intense than that in the osteoclast-like multinuclear giant cells. Some spherical mononuclear cells (macrophage-like cells) expressed high levels of PAI-1 mRNA in comparison with the spindle-shaped mononuclear cells. In addition, the 35th passaged spindle-shaped mononuclear cells were used to study the gene expression of u-PA during cell proliferation. The results showed that the level of u-PA mRNA increases after adding 10% fetal calf serum to quiescent cells. The induction was maximal at 16 hours and remained high during 48 hours of treatment. In conclusion, even though osteoclast-like cells are ultimately responsible for the bone resorption of GCT, the mononuclear neoplastic cells of GCT may also be involved in degradation of the extracellular matrix during invasive growth by facilitating the urokinase plasminogen activation system. In addition, our observation of upregulation of u-PA mRNA in spindle shaped mononuclear cells after serum stimulation indicated that u-PA production may be linked to tumor growth. PMID- 7495281 TI - Apolipoprotein(a) deposition in atherosclerotic plaques of cerebral vessels. A potential role for endothelial cells in lesion formation. AB - Atherosclerosis is the leading cause of death and serious morbidity in economically developed societies through its sequelae of coronary artery and cerebrovascular disease. The causes and mechanisms of atherosclerosis are still largely unknown. Serum levels of a lipoprotein, Lp(a), have been shown, in retrospective and some prospective clinical studies, to be associated with increased risk of myocardial and cerebral infarction. The active part of Lp(a), apo(a), has > 80% homology with plasminogen; thus it may competitively inhibit the thrombolytic action of plasminogen and enhance thrombogenesis. Lp(a) has been shown to be deposited in the vascular wall of the aorta and coronary vessels, but its presence in the cerebral vessels has not yet been shown. Autopsy specimens of vessels of the circle of Willis from 23 patients were examined for degree of atherosclerosis and deposition of apo(a) by immunohistochemistry with apo(a) specific monoclonal antibodies. The amount of apo(a) deposition in cerebral vessels correlated well with the degree of cerebral atherosclerosis. Arterial deposition of apo(a) was found entirely within the endothelial cell and subendothelial cell layers. There was no staining within the media and adventitia, with the exception of staining within the endothelial cells of the vasa vasorum. Correlation between the morphology of apo(a) deposition and plaque stage was found suggesting that detection of apo(a) in endothelial cells is an early event in the development of the atherosclerotic plaque of cerebral vessels. PMID- 7495282 TI - Evidence of apoptotic cell death after experimental traumatic brain injury in the rat. AB - Apoptosis plays an important role in many developmental and pathological processes of the central nervous system. However, the role of apoptosis in traumatic brain injury has not been determined. Using the terminal deoxynucleotidyl transferase-mediated biotinylated deoxyuridine triphosphate nick end labeling (TUNEL) method, we detected many cells with extensive DNA fragmentation in different regions of the brains of rats subjected to experimental traumatic brain injury. Two types of TUNEL-positive cells were demonstrated by light and electron microscopy, including type I cells that displayed morphological features of necrotic cell death and type II cells that displayed morphological features of classic apoptotic cell death. TUNEL-positive cells were detectable for up to 72 hours after the initial injury. Gel electrophoresis of DNA extracted from affected areas of the injured brain containing both type I and II cells revealed only internucleosomal fragmentation at 185-bp intervals, a feature originally described in apoptotic cell death. These data suggest that apoptosis, in addition to necrotic cell death, occurs after traumatic brain injury, and that internucleosomal fragmentation of DNA may be associated with certain types of necrotic cell death. PMID- 7495283 TI - Detection of chimeric transcripts in desmoplastic small round cell tumor and related developmental tumors by reverse transcriptase polymerase chain reaction. A specific diagnostic assay. AB - Desmoplastic small round cell tumor is a recently described entity associated with fusion of the EWS and WT1 genes and with expression of a chimeric transcript. To investigate the structure and potential diagnostic utility of the detection of EWS-WT1 chimeric RNA in desmoplastic small round cell tumor, 12 examples of this entity and 49 other tumors that enter in its differential diagnosis were studied by reverse transcriptase polymerase chain reaction for the presence of EWS-WT1, EWS-FLI-1, PAX3-FKHR, and PAX7-FKHR chimeric transcripts. EWS-WT1 was detected in 11 of 12 desmoplastic small round cell tumors but not in any other tumor type studied, including 17 Wilms' tumors, 10 Ewing's sarcomas/primitive neuroectodermal tumors, 13 alveolar rhabdomyosarcomas, and 9 embryonal rhabdomyosarcomas. One desmoplastic small round cell tumor was found to have a variant EWS-WT1 chimeric product that included exon 8 of EWS EWS-FLI-1 chimeric RNA was present in all Ewing's sarcoma/primitive neuroectodermal tumor and not identified in any other tumor types, including desmoplastic small round cell tumor. PAX3/PAX7-FKHR chimeras were present in 9 of 13 alveolar rhabdomyosarcomas but not in any other tumors. Detection of chimeric transcripts by reverse transcriptase polymerase chain reaction is a very specific aid in differential diagnosis of developmental tumors and further establishes desmoplastic small round cell tumor as a distinct entity. PMID- 7495284 TI - Molecular diagnosis of synovial sarcoma and characterization of a variant SYT SSX2 fusion transcript. AB - The translocation t(X;18)(p11;q11) is seen in > 80% of synovial sarcomas (SS) with informative karyotypes. The breakpoints of the t(X;18) have been cloned and shown to involve two novel genes, SSX (at Xp11) and SYT (at 18q11), which produce a chimeric SYT-SSX transcript as a result of the translocation. Recently, SSX has been shown to be duplicated, with both copies, SSX1 and SSX2, located within distinct subregions of Xp11. We performed a reverse transcriptase polymerase chain reaction (RT-PCR) assay for both chimeric SYT-SSX transcripts in a series of 35 SS (29 monophasic, 6 biphasic) to assess its usefulness in molecular diagnosis and to evaluate the incidence of molecular variants. Of the 35 cases, 29 (83%) showed a specific SYT-SSX RT-PCR product, using a consensus primer for SSX1 and SSX2 Upon excluding three negative cases that had poor quality RNA, the proportion of positives rose to 91% (29/32). The 29 positive cases were further studied using primers specific for either SSX1 or SSX2; 19 cases were positive for SYT-SSX1 and 10 for SYT-SSX2. The relationship of histological subtype (monophasic versus biphasic) to SSX1 or SSX2 involvement was not statistically significant. In a single histologically unremarkable monophasic SS, a slightly larger SYT-SSX2 RT-PCR product was observed. Sequencing of this novel variant showed a 129-bp segment inserted between the usual SYT and SSX2 fusion points, of which 126 bp were derived from a more proximal (5') portion of SSX2 The 3 bp immediately 5' to the fusion point could not be assigned to either SYT or SSX2 and may represent an insertion-deletion or a cryptic splicing event. This fragment maintains the reading frame of the chimeric product and encodes a predicted protein larger by 43 amino acids, which nevertheless replaces the region homologous to the transcriptional repression domain Kruppel-associated box, recently recognized in the 5' portion of the SSX genes, with all but the 3' end of the SYT transcript. Thus, a diagnosis of SS may be confirmed in > 90% of cases using RT-PCR detection of the chimeric transcript resulting from the t(X;18), and the incidence of molecular variants appears low. PMID- 7495285 TI - Absence of RET proto-oncogene point mutations in sporadic hyperplastic and neoplastic lesions of the parathyroid gland. AB - We investigated the possible role of RET proto-oncogene mutations in the development of sporadic hyperplastic, benign, and malignant parathyroid lesions. DNA extracted from paraffin-embedded specimens of forty parathyroid lesions was screened for RET proto-oncogene point mutations in exons 10, 11, and 16 by nonisotopic polymerase chain reaction-based single-strand conformation polymorphism and heteroduplex gel electrophoresis. The nucleotide sequence of samples with aberrant band patterns was identified by nonisotopic direct sequencing of polymerase chain reaction-amplified DNA. Parathyroids of seven patients with multiple endocrine neoplasia type 2A (MEN 2A) and MEN 2B served as positive controls. None of the eight hyperplastic lesions, three cases of parathyromatosis, ten parathyroid adenomas, eleven carcinomas or one normal parathyroid gland contained mutations in each of the three RET exons tested. Six MEN-2A-associated hyperplastic glands exhibited identical band shifts in the polymerase chain reaction single-strand conformation polymorphism analysis of exon 11, which corresponded to a Cys 634-->Arg substitution in the nucleotide sequence analysis (TGC-->CGC), whereas in the MEN 2B parathyroid specimen a point mutation was found at codon 918 of exon 16 (ATG-->ACG), causing a Met 918-->Thr substitution. Our data indicate that RET mutations of the MEN 2 loci in exons 10, 11, and 16 are not involved in the development of sporadically occurring benign or malignant parathyroid lesions. Furthermore, our results are in accordance with the observation that MEN 2A patients with Cys 634-->Arg (germline) mutations have a higher risk of developing parathyroid disease than those with other mutations at codon 634. PMID- 7495286 TI - Analysis of genetic changes underlying local recurrence of prostate carcinoma during androgen deprivation therapy. AB - The molecular mechanisms and genetic changes that lead to the progression of prostate cancer during endocrine therapy are poorly characterized. Here, paired specimens from both untreated primary tumors and from local recurrences were collected from 10 prostate cancer patients treated by conventional androgen deprivation therapy. The genetic progression of the tumors was studied by using interphase fluorescence in situ hybridization and chromosome-specific probes. Six primary tumors (60%) and all ten recurrent tumors were aneuploid by fluorescence in situ hybridization. The recurrent tumors also showed a high degree of chromosome copy number variability from one cell to another. Increased copy number of chromosome X was particularly common in the recurrent tumors. In addition, specific high level amplification of the androgen receptor (AR) gene (Xq12) was detected in three highly aneuploid recurrent tumors. Our findings suggest that hormone-refractory prostate cancers are genetically very complex and show intratumor genetic heterogeneity. Increased copy number of chromosome X and the amplification of the androgen receptor (AR) gene may confer proliferative advantage during androgen deprivation and thus contribute to the development of recurrence. PMID- 7495287 TI - Diagnostic assessment of two novel proliferation-specific antigens in benign and malignant melanocytic lesions. AB - The aim of this study was to gain a thorough insight into the proliferative activity of benign and malignant melanocytic tumors. A total of 314 cases were examined by immunohistochemistry on paraffin-embedded material. The growth fraction was assessed by means of two monoclonal antibodies, Ki-S1 and Ki-S5, which react with two different proliferation-specific nuclear antigens. Additionally, HMB-45 was used as a marker of melanocytic activation. Statistically significant differences (P < 0.01) in the proliferation rates were found between common acquired nevi, Spitz's/Reed's nevi, primary cutaneous melanomas, and metastatic melanomas, whereas dysplastic nevi were hardly distinct from other nevi of the compound type. In melanoma, the growth fraction correlated well with the tumor stage but poorly with HMB-45 expression and mitotic count. Along with tumor progression, an increasing heterogeneity of proliferation indices was observed. Our results provide no evidence for a progression from dysplastic nevi into melanoma. They indicate that the assessment of the proliferative activity may be of considerable diagnostic help in cases of uncertain histology and that it might contribute to an alternative concept for the classification of melanocytic tumors. PMID- 7495288 TI - Quantitative in situ image analysis of apoptosis in well and poorly differentiated tumors from rat liver. AB - The evaluation of apoptosis is an important aspect in the study of chemical carcinogenesis. Methods were developed employing the ApopTag kit (Oncor, Gaithersburg, MD) and automated image analysis to quantitate the distribution of apoptosis in formalin-fixed, paraffin-embedded liver tumor sections from rats induced by 2-acetylaminofluorene. Specific treatments of tissue sections were developed that permitted quenching of background tissue autofluorescence with crystal violet and permitted permeating the fixed tissue sections by trypsin digestion. Tissue sections were stained by using the ApopTag kit for detection of in situ apoptosis and with propidium iodide as a counterstain for tissue nuclei. Automated statistical evaluation of the percentage of tissue nuclei also staining positively for apoptosis was determined by using dual fluorescence detection and imaging laser microscopy. The quantitative results indicated that the staining index for apoptosis in normal liver was 0.14 +/- 0.04% whereas well and poorly differentiated tumors showed increases of 3.48 +/- 0.59% and 7.41 +/- 0.81%, respectively. The staining indexes for apoptosis showed a tight correlation between fluorescent and peroxidase-diaminobenzidine detection in sequential tissue sections. The use of in situ apoptosis staining and automated image analysis for rapid identification and quantitation of cells undergoing death in fixed tissue will expedite additional studies to evaluate the in situ tissue pathobiology of tumors and aid in the study of molecular mechanisms associated with cancer development and treatment. PMID- 7495289 TI - Transcription factor and liver-specific mRNA expression in facultative epithelial progenitor cells of liver and pancreas. AB - The pattern of mRNA expression for liver-specific proteins and liver-enriched transcription factors was studied in two models of facultative gut epithelial progenitor cells activation: D-galactosamine (GalN)-induced liver injury and dietary copper depletion leading to pancreatic acinar atrophy. After 5 weeks of copper deficiency (CuD), pancreatic acini of Fischer 344 rats underwent atrophy, associated with intense proliferation of small duct-like cells with oval-shaped nuclei. These cells resemble morphologically epithelial progenitor cells of the liver that proliferate after GalN administration. Activated pancreatic epithelial cells express mRNAs for liver-specific genes normally expressed in fetal liver, including alpha-fetoprotein, albumin, alpha-1 antitrypsin, glucose-6-phosphatase, and others, but not genes that are turned on after birth such as serine dehydratase, tyrosine aminotransferase, and multidrug resistance gene-1b. They express mRNAs for liver-enriched transcription factors including HNF-1 alpha, HNF 3 beta and gamma, HNF-4, and members of the CCAAT-enhancer binding protein (C/EBP) family. The only mRNA for a liver-enriched transcription factor not detected in the pancreas of CuD animals was HNF-3 alpha. Expression of HNF-3 alpha, beta, and gamma, and C/EBP-beta mRNA was highly activated in proliferating liver epithelial cells on days 2 and 3 after GalN injury. Increased expression of C/EBP-delta was observed first in the liver on day 1 after GalN administration and in the pancreas at 4 weeks after initiating CuD. We suggest that C/EBP-delta could be involved in the initial activation of epithelial progenitor cells and that HNF-3 alpha, beta, and gamma, and C/EBP-beta might participate in their maturation. We conclude further that pancreatic epithelial progenitor cells undertake differentiation through the hepatocyte lineage but cannot complete the differentiation program within the pancreatic milieu. PMID- 7495290 TI - Time course of increased cellular proliferation in collateral arteries after administration of vascular endothelial growth factor in a rabbit model of lower limb vascular insufficiency. AB - Proliferation of vascular cells has been previously shown to contribute to spontaneous development of coronary collaterals. Recent studies from several laboratories have established that collateral artery growth in both the heart and limb can be enhanced by administration of angiogenic growth factors, or therapeutic angiogenesis. In this study, we sought (1) to define the extent and time course of endothelial cell (EC) and smooth muscle cell (SMC) proliferation accompanying spontaneous collateral development during limb ischemia and (2) to determine the extent to which proliferative activity of ECs and SMCs is augmented during therapeutic angiogenesis with vascular endothelial growth factor (VEGF), a heparin-binding EC-specific mitogen. Ten days after induction of limb ischemia by surgically excising the femoral artery of rabbits, either VEGF (500 to 1000 micrograms) or saline was administered as a bolus into the iliac artery of the ischemic limb. Cellular proliferation was evaluated by bromodeoxyuridine labeling for 24 hours at day 0 (immediately before VEGF administration) and at days 3, 5, and 7 after VEGF, EC proliferation in the midzone collaterals of VEGF-treated animals increased 2.8-fold at day 5 (P < 0.05 versus control), and returned to baseline levels by day 7. SMC proliferation in midzone collaterals also increased 2.7-fold in response to VEGF (P < 0.05). No significant increase in EC or SMC proliferation was observed in either the stem or re-entry collaterals of VEGF treated animals compared with untreated ischemic control animals. Reduction of hemodynamic deficit in the ischemic limb measured by lower limb blood pressure was documented at day 7 after VEGF (P < 0.01 versus untreated, ischemic control). These data thus (1) establish the contribution of cellular proliferation to collateral vessel development in limb ischemia and (2) support the concept that augmented cellular proliferation contributes to the enhanced formation of collateral vessels after therapeutic angiogenesis with VEGF. PMID- 7495291 TI - Altered vascular endothelium integrin expression in psoriasis. AB - Considerable evidence indicates that microvascular changes observed in psoriasis are a result of vascular proliferation. A critical step in the sequence of events leading to neovascularization involves interactions between endothelial cells and extracellular matrix proteins mediated in part by the integrin family of adhesion molecules. A number of endothelial integrins have been shown to participate in neovascularization, including members of the beta 1, beta 3, and beta 4 subfamilies. To investigate the role of these integrins in psoriasis, specimens of lesional and nonlesional skin were taken from 10 patients with active, untreated plaque disease. Vascular endothelium was labeled with monoclonal antibodies specific for alpha 2, alpha 5, alpha 6, beta 1, av beta 3, and beta 4 integrins. The use of image analysis permitted quantification of immunoperoxidase staining and comparison of endothelial labeling in lesional and nonlesional skin. There was a significant increase in endothelial staining of av beta 3 integrin in lesional compared with nonlesional skin, both in superficial and deep vasculature. In contrast, there was a significant decrease in endothelial beta 4 staining in lesional compared with nonlesional superficial dermal vessels, alpha 2, alpha 5, alpha 6, and beta 1 staining showed no significant difference between the two groups. These results demonstrate an important role of av beta 3 and beta 4 integrins in the microvascular changes of psoriatic lesions. PMID- 7495292 TI - Highly metastatic 13762NF rat mammary adenocarcinoma cell clones stimulate bone marrow by secretion of granulocyte-macrophage colony-stimulating factor/interleukin-3 activity. AB - Circulating neutrophil (polymorphonuclear leukocyte levels rise 50-fold in 13762NF tumor-bearing rats in proportion to the tumor's metastatic potential. Purified tumor-elicited neutrophils enhance metastasis of syngeneic tumor cells when co-injected intravenously; however, circulating and phorbol ester-activated polymorphonuclear neutrophils do not. The purpose of this study was to elucidate the source of tumor-elicited neutrophils in metastatic tumor-bearing rats. We examined the bone marrow in rats bearing tumors of poorly, moderately, and highly metastatic cell clones. Marrow from rats with highly metastatic tumors had increased cellularity (100%), myeloid to erythroid ratio (10:1), and megakaryocytes compared with control rats (cellularity, approximately 80%; myeloid to erythroid ratio, 5:1), with marrows from rats with moderately metastatic tumors having intermediate values. This suggested production of a colony-stimulating factor by the metastatic cells. To confirm this, bone marrow colony formation from control and tumor-bearing rats was compared. Colony number increased in proportion to the metastatic potential of the tumor. Conditioned medium from metastatic cells supported growth of the granulocyte-macrophage colony-stimulating factor/interleukin-3-dependent 32Dcl3 cell line, but media from nonmetastatic or moderately metastatic cells did not. Antibodies to murine granulocyte-macrophage colony-stimulating factor neutralized 32Dcl3 growth in tumor cell conditioned medium. These results suggest production of a granulocyte macrophage colony-stimulating factor or interleukin-3-like activity by highly metastatic 13762NF clones and implicate a possible role for colony-stimulating factors in regulating the metastatic potential of mammary adenocarcinoma cell clones. PMID- 7495293 TI - Bone marrow neutrophilia and suppressed bone turnover in human interleukin-6 transgenic mice. A cellular relationship among hematopoietic cells, osteoblasts, and osteoclasts mediated by stromal cells in bone marrow. AB - To elucidate the effect of interleukin-6 (IL-6) on bone and bone marrow (BM), human IL-6 transgenic mice (hIL-6 tgm) were produced. Their bone and BM were examined histologically, radiologically, histomorphometrically, and hematologically on a temporal basis. hIL-6 tgm showed histologically evident neutrophilia in BM. Increase in precursors of granulocytes and monocytes in hIL-6 tgm was demonstrated by an assay for colony forming unit in culture (CFU-C) of BM cells. Decrease in osteoblasts and osteoid and suppression of primary spongiosa formation were predominantly observed in hIL-6 tgm at 14 weeks old, the terminal stage of life for hIL-6 tgm. An assay for colony forming unit in fibroblastic (CFU-F) of BM cells revealed a decrease in osteoblast precursor (with regard to alkaline phosphatase-positive colonies) in hIL-6 tgm at 15 weeks old. Histomorphometry demonstrated a decrease of both osteoclast number and bone resorption in hIL-6 tgm. These results suggested that enhanced granulocytic hematopoiesis, suppressed bone turnover, and alteration of cellular population in stromal cells in BM occurred in hIL-6 tgm. Thus we provide new findings that facilitate understanding of cellular interrelationships among hematopoietic cells, osteoblasts, and osteoclasts mediated by stromal cells in BM. PMID- 7495294 TI - Role of interferon-gamma in interleukin 12-induced pathology in mice. AB - Interleukin 12 (IL-12) activates natural killer (NK) and T cells with the secondary synthesis and release of interferon-gamma (IFN-gamma) and other cytokines. IL-12-induced organ alterations are reported for mice and the pathogenetic role of IFN-gamma is investigated by the use of mice deficient in the IFN-gamma receptor (IFN-gamma R-/-). IL-12 caused a rapid infiltration of liver and splenic red pulp with activated macrophages; this and increased NK cells resulted in a fivefold increase of splenic weight in wild-type mice. Splenomegaly was associated with myelosuppression and decreasing peripheral leukocyte counts. IL-12-induced changes in wild-type mice were associated with markedly increased IFN-gamma serum levels and up-regulation of major histocompatibility complex (MHC) class I and II expression in various epithelia. IL-12 induced a qualitatively similar macrophage infiltration in IFN-gamma R-/- mice, less marked splenomegaly (to 2 x normal), and no MHC upregulation. Strikingly increased vascular endothelial intercellular adhesion molecule-1 expression was apparent in both IFN-gamma R-/- and IFN-gamma R+/+ mice. Restricted to mutant mice was a severe, invariably lethal, interstitial, and perivascular pulmonary macrophage infiltration with diffuse pulmonary edema. Extensive quantitative reverse transcriptase polymerase chain reaction analysis revealed an increase of only IL-6 and IL-10 pulmonary gene transcripts in IFN gamma R-/- mice compared with wild-type mice. IL-12-induced myelosuppression is due to IFN-gamma-release from NK cells and T cells, and is associated with macrophage activation and distinct MHC class I and II antigen upregulation. The pulmonary pathology in IFN-gamma R-/- mice, however, reveals a toxic potential for IL-12 and suggests that endogenous IFN-gamma plays a protective role in preventing fatal pulmonary disease in these mice. PMID- 7495295 TI - Peripheral blood leukocyte grafts that induce human to mouse graft-vs.-host disease reject allogeneic human skin grafts. AB - Allogeneic graft-versus-host disease is characterized by skin, gut, and bile duct destruction by relatively few donor type lymphocytes. In contrast, we can now show that human-to-mouse xenogeneic graft-versus-host disease is characterized by vasculitis and tumor-like infiltrations of the murine lymphohemopoietic organs with many human CD25+, HLA-DR+, CD4+ lymphoblasts. Using the technique of serial transplantation, it appears that at least 90% of the human lymphoblasts were unreactive to murine tissues. It is demonstrated consistently that the donor type lymphoblasts induced typical allogeneic rejection of distantly located full thickness human unmatched fetal skin grafts. The fact that the human grafts show primary immune responses in vivo indicates that the graft-versus-host disease murine model may be suitable for vaccination studies. PMID- 7495296 TI - Participation of glomerular endothelial cells in the capillary repair of glomerulonephritis. AB - In many glomerular diseases severe injury to the mesangium may occur, leading to matrix dissolution and damage to the glomerular capillaries. Although the destruction of glomerular architecture may lead to permanent injury, in some cases spontaneous recovery occurs. The mechanisms that mediate this recovery are unknown. In this study we provide evidence for glomerular capillary repair (angiogenesis) in the adult injured glomerulus. Injection of anti-Thy 1 antibody into rats results in severe mesangiolysis with capillary ballooning, microaneurysm formation, and loss of endothelial cells in addition to mesangial cells. Although mesangial proliferation is a major response to injury, proliferation of endothelial cells also can be documented from days 2 to 14 in association with repair of the capillaries. The endothelial cell proliferation peaks on days 2 and 7, when it is seven- to ninefold greater than normal. Many of the endothelial cells display morphological features of angiogenesis. The initial wave of endothelial cell proliferation can be reduced by 40% with neutralizing anti-basic fibroblast growth factor antibodies (P < 0.001). The later glomerular endothelial cell proliferation is associated with upregulated expression of vascular permeability factor/endothelial cell growth factor (VPF/VEGF) and an increase of flk, a VPF/VEGF receptor. Although PDGF is expressed in this model, anti-PDGF antibody treatment did not affect the endothelial cell proliferative response. In summary, glomerular endothelial cells have an active role in the glomerular response to injury. Glomeruli are capable of healing microaneurysms, and the mechanism involves basic fibroblast growth factor- and VPF/VEGF-mediated endothelial proliferative responses. PMID- 7495297 TI - Analysis of the role of membrane polarity in polycystic kidney disease of transgenic SBM mice. AB - Altered membrane polarity has been proposed as an important pathogenetic factor in the development of renal cysts in polycystic kidney disease. To determine whether this alteration in epithelial phenotype is a primary or secondary phenomenon, we examined the epithelial membrane polarity of SBM transgenic mice, in which epithelial proliferation mediated by the c-myc oncogene is an established primary event. Kidneys from 32 transgenic mice and 10 age-matched controls from fetal to adult age were immunostained with antibodies to Na,K ATPase, fodrin, ankyrin, E-cadherin, and tubule segment-specific lectins. In normal control mice, Na,K-ATPase localization was apical in fetal kidneys but became translocated to the basolateral membrane at maturity. Early microcysts in fetal transgenic kidneys displayed similar (95 to 100%) apical Na,K-ATPase. In young and newborn transgenic mice (1 to 8 days of age), Na,K-ATPase localization was extremely heterogeneous. Noncystic tubules demonstrated either apical (mean 23 to 28%), basolateral (mean 48 to 58%), mixed (mean 4 to 15%), or absent (mean 10 to 13%) staining for Na,K-ATPase. Apical Na,K-ATPase was more frequently observed in early cysts (mean 55%) in young transgenic mice but became less prevalent in adult mice (mean 22%), where 30% of cysts had basolateral staining, 39% mixed patterns, and 9% absent staining. Macrocysts typically lost all Na,K ATPase reactivity. At all ages, Na,K-ATPase colocalized well with cytoskeletal proteins ankyrin and fodrin. These heterogeneous patterns of Na,K-ATPase staining indicate that although altered cell polarity is frequent in early cystic epithelium of SBM mice, it is not a prerequisite to cystogenesis or progressive cyst enlargement. In conclusion, our results support the view that altered cystic membrane polarity is not a primary process, but represents the persistence of an immature epithelial phenotype characteristic of proliferative polycystic kidney disease epithelia. PMID- 7495298 TI - Glomerular tissue factor expression in crescentic glomerulonephritis. Correlations between antigen, activity, and mRNA. AB - Correlations between glomerular expression of tissue factor (TF) activity and antigen and cellular localization of TF mRNA was studied in crescentic glomerulonephritis (GN) in rabbits. Glomerular TF activity increased 8.7-fold 24 hours after initiation of GN (234 +/- 49 mU/10(3) glomeruli; normal, 27 +/- 10 mU/10(3) glomeruli; P = 0.003) in association with a 2.1-fold increase in TF antigen (154 +/- 34 ng/10(3) glomeruli; normal, 72 +/- 10 ng/10(3) glomeruli; P = 0.055), early macrophage infiltration, and no significant increase in TF mRNA. At the peak glomerular macrophage infiltration (day 4), TF activity remained augmented (230 +/- 63 mU/10(3) glomeruli) and TF mRNA, colocalized within macrophages, was significantly increased compared with normal (267 +/- 42%; P = 0.001). TF antigen was not increased in glomeruli (114 +/- 17 ng/10(3) glomeruli), although significant urinary excretion of TF antigen was detectable (478 +/- 121 ng/24 hours; normal, < 1 ng/24 hours; P = 0.032). At this time, the M(r) of glomerular TF (49 to 61 kd) was increased compared with TF in normal glomeruli (49 to 58 kd) as a result of increased glycosylation. At day 7, TF activity and antigen within glomeruli had decreased, although urinary excretion of TF antigen and glomerular TF mRNA remained elevated. These studies suggest that early up-regulation of TF activity is largely a result of functional up regulation of constitutive TF in intrinsic glomerular cells. In more advanced disease, infiltrating macrophages are the major site of TF synthesis. The increased M(r) of glomerular TF, as a result of synthesis of more highly glycosylated protein by macrophages and the shedding of TF into the urine, suggests that substantial turnover of glomerular TF occurs at this stage. PMID- 7495299 TI - Chronic alcohol feeding inhibits atherogenesis in C57BL/6 hyperlipidemic mice. AB - Although there is abundant clinical evidence that the consumption of alcohol (ethanol) in moderate amounts has a protective effect on coronary artery disease, the mechanism of this effect is not understood. The prevailing theory supported by a limited number of clinical and experimental animal studies indicates that the ability of alcohol to elevate serum high-density lipoprotein cholesterol levels is an important mechanism. Although there have been a large number of studies on the effects of alcohol on serum lipoprotein and apolipoproteins on coronary artery disease, there have been very few that have, at the same time, looked directly and systematically at its effects on the histopathological development of atherosclerotic lesions. In the following studies we employed the hyperlipidemic C57BL/6 female mouse model and formulated an all liquid high fat atherogenic diet to provide the mice with the 3% or 6% alcohol. After 22 weeks on this diet, alcohol markedly inhibited the development of fatty streak atherosclerotic lesions in a dose-dependent fashion. Surprisingly, there was a dose-dependent decrease in plasma high-density lipoprotein cholesterol values, which suggests that high-density lipoprotein alterations play little or no role in the amelioration of atherosclerosis in this model. PMID- 7495300 TI - Immunolocalization of SPARC, tenascin, and thrombospondin in pulmonary fibrosis. AB - Several biochemically unrelated multifunctional extracellular proteins, SPARC, thrombospondin 1, and tenascin-C (TN), have been grouped as antiadhesive glycoproteins because they inhibit the spreading of cells on extracellular matrix in vitro. Migration of fibroblasts and epithelial cells into the air spaces to organize inflammatory exudate is a feature common to several fibrosing lung diseases. We hypothesized that migration would be facilitated by loosening the adhesive interactions between cells and the pericellular matrix components of the alveolar wall and that one or more of the anti-adhesive glycoproteins could be involved. Immunohistochemistry was used to localize SPARC, TN, and thrombospondin 1 in biopsies of organizing pneumonia, idiopathic pulmonary fibrosis (nine cases of usual interstitial pneumonia, one of desquamative interstitial pneumonia), and control lungs. Each antigen had a distinctive distribution. Only TN was expressed in control lungs, where it strongly stained the basement membrane of large bronchi and weakly stained alveolar entrance rings and small veins. In organizing pneumonia, TN was heavily stained through the entire extracellular matrix of the Masson bodies. In idiopathic pulmonary fibrosis, TN was abundant in the fibroblast foci of active fibrosis but was also present in the basement membrane regions beneath the metaplastic epithelium lining honeycomb cysts. TN was abundant in the interstitium in desquamative interstitial pneumonia. SPARC was observed only intracellularly where it occurred in the fibroblasts of Masson bodies of organizing pneumonia and the fibroblast foci of usual interstitial pneumonia. In desquamative interstitial pneumonia, expression of SPARC was minimal, in rare interstitial fibroblasts. Thrombospondin 1 was found consistently in organizing pneumonia but only infrequently in idiopathic pulmonary fibrosis. In both, it was localized in the extracellular matrix immediately beneath reparative epithelium. These results are consistent with a role for SPARC in fibroblast migration. TN may function in both fibroblast migration and the adhesion of metaplastic bronchial-type epithelium. However, these proteins also have other activities that may be important in pulmonary fibrosis. The localization of thrombospondin 1 suggests that it may be synthesized by regenerating epithelium where it may aid in the adhesion or migration of the epithelial cells. PMID- 7495301 TI - DNA aberrations in the epithelial cell component of adamantinoma of long bones. AB - Adamantinoma of long bones is a rare malignant tumor composed of cells with epithelial characteristics in various differentiation patterns surrounded by fibrous cells. Evidence as to whether this neoplasm should be designated as an epithelial bone tumor or a biphasic sarcoma with both epithelial and mesenchymal features is lacking. In this study the nature of the mesenchymal and epithelial components of adamantinoma was investigated by DNA flow cytometry, DNA image cytometry, p53 immunohistochemistry, and polymerase chain reaction-based loss of heterozygosity detection at the p53 locus. Specimens from 6 of 15 patients (40%) analyzed by flow cytometry had an aneuploid DNA index. Image cytometry analysis of Feulgen-stained paraffin sections of 6 aneuploid and 2 diploid tumors revealed that aneuploid nuclei were detected in cells with an epithelial phenotype only, whereas all fibrous cells were diploid. Immunohistochemistry for p53 on specimens from 25 patients revealed moderate or strong immunoreactivity in 12 tumors (48%) restricted to the epithelial cells. Loss of heterozygosity at the p53 locus could be confirmed in the epithelial component of an immunohistochemically p53-positive tumor. Additionally, sections of 7 lung metastases were studied histologically. Only keratin-positive epithelial cells, predominantly in the spindle cell pattern, were present in these metastases, whereas the osteofibrous tissue present in the primary tumors was not detected. These results suggest that either adamantinoma consists of a malignant epithelial part with a reactive osteofibrous stroma or that the malignant epithelial cells develop next to a proliferating benign fibrous component. Additional analysis of common genetic abnormalities in the fibrous and epithelial cells of adamantinoma is therefore indicated. PMID- 7495302 TI - HIV-1 gp160 protein-macrophage interactions modulate mesangial cell proliferation and matrix synthesis. AB - Patients with HIV infection often develop glomerular lesions (focal segmental glomerular sclerosis). Because mesangial expansion (enhanced mesangial cell (MC) growth and matrix accumulation) has been demonstrated to precede the development of focal segmental glomerulosclerosis, we studied the effect of the interaction between HIV-1 proteins such as gp160 envelope protein and macrophages on mesangial cell proliferation and matrix synthesis. We determined the effect of control media, serum-free macrophage supernatant (MSP), and serum-free HIV-1 gp 160 protein-treated MSP (gp 160-MSP) on the proliferation of MC and synthesis of collagen type IV (a component of mesangial matrix). MSP (20%) enhanced (P < 0.01) MC proliferation (control, 7.58 +/- 0.29 versus MSP, 9.06 +/- 0.25 x 10(4) cells/ml), whereas gp 160-MSP (20%) inhibited (P < 0.001) MC proliferation (gp160 MSP, 5.58 +/- 0.14 x 10(4) cells/ml). gp160-MSP modulated MC proliferation in a dose-dependent manner; it enhanced cell proliferation at a lower concentration but inhibited cell proliferation at a higher concentration. Anti-TGF-beta antibody attenuated the effect of gp160-MSP on MC proliferation at lower as well as higher concentrations. Bromodeoxyuridine incorporation studies also showed the modulation of MC proliferation by gp160-MSP. Interaction of other HIV proteins such as HIV-1 Gag4 and HIV-1 Tat with macrophages did not affect MC proliferation when compared with MSP alone. gp160-MSP also enhanced (P < 0.001) synthesis of type IV collagen by MC (control, 467.8 +/- 9.0; MSP, 501.0 +/- 25.0; gp160-MSP, 775.5 +/- 39.0 ng/mg protein). The effect of gp160-MSP on collagen synthesis by MC was dose-dependent. Anti-TGF-beta antibody attenuated the gp160-MSP-induced mesangial cell collagen synthesis. The present study provides a basis for speculation that macrophage-gp160 interaction products have the potential to cause expansion of the mesangium. PMID- 7495304 TI - Expression of myogenic regulatory proteins (myogenin and MyoD1) in small blue round cell tumors of childhood. AB - The distinction of rhabdomyosarcoma (RMS) from other small blue round cell tumors of childhood, such as Ewing's sarcoma/peripheral primitive neuroectodermal tumor (pPNET) and neuroblastoma, continues to present a diagnostic challenge to pathologists. The recent recognition of the master role of myogenic regulatory proteins in skeletal muscle commitment and differentiation, and the availability of monoclonal antibodies to two of them (myogenin and MyoD1), has prompted us to test their diagnostic utility in routinely processed, formalin-fixed, and deparaffinized tissue. Preliminary studies had demonstrated that, with the use of heat-induced epitope retrieval techniques, expression of myogenin and MyoD1 could be documented specifically in nuclei of fetal skeletal muscle by the respective antibodies. We performed a retrospective immunohistochemical analysis on 72 cases of small blue round cell tumors, including 33 RMSs, 1 metastatic myogenous Wilms' tumor, 26 Ewing's sarcomas/pPNETs, and 12 neuroblastomas. Nuclear expression of myogenin and MyoD1 were both found in 30/33 non-overlapping cases of RMS, with no significant differences in the sensitivity with respect to histological subtypes, and in 1/1 case of myogenous Wilms' tumor. None of the neuroblastomas or Ewing's sarcomas/pPNETs demonstrated positive nuclear staining with either antibody. However, most of the neuroblastomas, and occasional Ewing's sarcomas/pPNETs, showed variable fibrillary, cytoplasmic immunoreactivity with antibody to MyoD1. We conclude that, with the use of microwave-based epitope retrieval, antibodies to myogenin and MyoD1 are both useful markers for the identification of RMS among other small blue round cell tumors of childhood, but antibodies to myogenin have technical advantages over those to MyoD1, as the latter may cross-react with an unknown cytoplasmic antigen in non-muscle cells and tumors. PMID- 7495303 TI - Insulin-like growth factor II in the pathogenesis of human neuroblastoma. AB - Insulin-like growth factor II (IGF-II) acts as an autocrine growth factor for many in vitro tumor cell lines including neuroblastoma. To examine the role of IGF-II in tumor biology we have analyzed a total of 56 primary neuroblastoma tumor samples for the presence of IGF-II using a combination of mRNA and protein analysis. A group of 21 samples was examined for the presence of IGF-II mRNA by slot blot and a separate group of 37 samples was examined for IGF-II immunoreactivity. IGF-II was detected in 48% of the total tumor specimens analyzed. IGF-II immunoreactivity was observed in cells resembling developing neuroblasts and was confined to the cytoplasm and proximal neurites. The appearance of IGF-II mRNA and protein did not correlate with tumor prognostic features including stage, histology, or N-myc amplification. These data suggest that the expression of IGF-II is not confined to a specific stage of the disease but may have a broader role in the pathogenesis of neuroblastoma. PMID- 7495305 TI - Conditional transformation of mouse liver epithelial cells. An in vitro model for analysis of genetic events in hepatocarcinogenesis. AB - Primary rodent and human hepatocytes have a very limited lifespan in culture and are not readily applicable to transformation studies in vitro. To facilitate the investigation of early genetic events involved in hepatocarcinogenesis, we examined a transformation assay system utilizing conditionally immortalized mouse liver epithelial cells as an alternative to primary hepatocytes. By infecting primary mouse hepatocytes with a recombinant retrovirus carrying a temperature sensitive simian virus 40 large T antigen gene, two mouse liver epithelial cell lines, CHST8 and CHST10-2.1, were established. Because of the heat-labile nature of the large T antigen, the cell lines proliferated rapidly at 33 degrees C, but were growth-arrested at 39 degrees C. Because activated c-H-ras and c-myc oncogenes are frequently found to be involved in mouse hepatocarcinogenesis in vivo, we assessed whether those oncogenes can complement the immortalizing function of the large T antigen at the nonpermissive temperature. When CHST8 cells were doubly transfected with activated c-H-ras and c-myc at 33 degrees C, they exhibited clonal growth ability even after shifting the temperature to 39 degrees C. However, neither c-H-ras nor c-myc alone allowed growth at 39 degrees C. On the other hand, c-H-ras alone was sufficient for overcoming the growth defect of CHST10-2.1 cells at 39 degrees C, whereas c-myc alone was again ineffective. Northern blot studies revealed that endogenous c-myc expression was significantly downregulated in the parental CHST8 cells after a temperature shift from 33 to 39 degrees C. In contrast, in the parental CHST10-2.1 cells, appreciable c-myc expression was observed at both temperatures. These results indicate that c-H-ras and c-myc can cooperate in complementing the ability of the temperature-sensitive large T antigen to immortalize mouse liver cells at the nonpermissive temperature. In addition, the mutant c-H-ras, but not c-myc, cooperated with the functional T antigen at 33 degrees C to allow growth in soft agarose of the CHST8 and CHST10-2.1 cell lines. However, cell lines carrying mutant c-H-ras and overexpressing c-myc were unable to grow in soft agarose at 39 degrees C. Thus, the two cellular oncogenes were insufficient for full transformation of the liver epithelial cells. The present in vitro model should be useful for investigating molecular events involved in both early and late stages of hepatocarcinogenesis. PMID- 7495306 TI - Expression of hemidesmosomes and component proteins is lost by invasive breast cancer cells. AB - Hemidesmosomes are multiprotein structures that attach basal cells of stratified epithelia to basement membranes. Although normal human breast epithelia are not stratified, we observed expression of electron-dense hemidesmosomes and hemidesmosome protein components by breast epithelial and myoepithelial cells at the basal lamina in vivo. Primary cultured normal human breast epithelial cells also contained hemidesmosomes and component proteins, and could be used as a model for hemidesmosome assembly and regulation. In these cultured cells, hemidesmosome proteins were expressed and localized basally in an unvaried temporal pattern, and electron-dense hemidesmosomes were not seen until the final protein was localized to the cell base. In addition, rate of localization was influenced by confluence, doubling time, and extracellular matrix. Invasive breast cancer cells did not express hemidesmosomes or most of the component proteins in vivo. In carcinoma in situ, cells away from the basement membrane lacked hemidesmosomes and hemidesmosome proteins, and cells at the basement membrane exhibited abnormalities of hemidesmosome protein expression. Primary human malignant breast cells in culture exhibited a mix of hemidesmosome phenotypes. These data suggest that hemidesmosomes may be important subcellular structures in determining the cytoarchitecture of the breast epithelium. Further, their downregulation may influence cytoarchitecture remodeling closely linked with cell cycle, motility, and extracellular matrix interactions; and their loss in carcinoma may be associated with loss of normal cytoarchitecture. PMID- 7495308 TI - Transfected type II interleukin-1 receptor impairs responsiveness of human keratinocytes to interleukin-1. AB - Of the two known types of specific receptors for interleukin (IL)-1, the function of the type II IL-1 receptor (IL-1RII) is still elusive. IL-1RII is allegedly devoid of signaling capacity and is therefore thought to act by trapping and inhibiting IL-1. To directly assess the functional role of IL-1RII, a human keratinocyte cell line has been stably transfected with a cDNA coding for IL 1RII, and its responsiveness to IL-1 has been compared with that of nontransfected cells. Parental cells express IL-1RI and are responsive to low doses of IL-1, whereas transfected cells overexpress IL-1RII, both in its membrane and soluble form, and show a dramatically impaired response to IL-1. Selective block of IL-1RII restores the ability of transfected keratinocytes to respond to IL-1, indicating that the overexpressed IL-1RII is in fact uniquely responsible for their refractoriness to IL-1. The main mechanism of unresponsiveness in transfected keratinocytes appears to be the capture and neutralization of IL-1 by the soluble form of IL-1RII. PMID- 7495307 TI - Comparison of intracerebral inoculation and osmotic blood-brain barrier disruption for delivery of adenovirus, herpesvirus, and iron oxide particles to normal rat brain. AB - Delivery of adenovirus, herpes simplex virus (HSV), and paramagnetic monocrystalline iron oxide nanoparticles (MION) to rat brain (n = 64) was assessed after intracerebral inoculation or osmotic disruption of the blood-brain barrier (BBB). After intracerebral inoculation, the area of distribution was 7.93 +/- 0.43 mm2 (n = 9) for MION and 9.17 +/- 1.27 mm2 (n = 9) for replication defective adenovirus. The replication-compromised HSV RH105 spread to 14.00 +/- 0.87 mm2 (n = 8), but also had a large necrotic center (3.54 +/- 0.47 mm2). No infection was detected when virus was administered intra-arterially without hyperosmotic mannitol. After osmotic BBB disruption, delivery of the viruses and MIONs was detected throughout the disrupted cerebral cortex. Positive staining was found in 4 to 845 cells/100 microns thick coronal brain section (n = 7) after adenovirus administration, and in 13 to 197 cells/section (n = 8) after HSV administration. Cells of glial morphology were more frequently stained after administration of adenovirus, whereas neuronal cells were preferentially stained after delivery of both HSV vectors and MION. In a preliminary test of vector delivery in the feline, MION was detected throughout the white matter tracts after inoculation into normal cat brain. Thus MION may be a tool for use in vivo, to monitor the delivery of virus to the central nervous system. Additionally, BBB disruption may be an effective method to globally deliver recombinant viruses to the CNS. PMID- 7495310 TI - Mild osteopetrosis in the microphthalmia-oak ridge mouse. A model for intermediate autosomal recessive osteopetrosis in humans. AB - Mutations at the mouse microphthalmia (mi) locus affect coat color, eye development, and mast cells. The original allele, mi, also shows severe osteopetrosis. Mice homozygous for the microphthalmia-Oak Ridge (Mior) mutation are white, microphthalmic animals with retarded incisor development. To investigate whether this mutation causes osteopetrosis, we examined skeletal tissues of the Mior mouse. A typical osteopetrotic lesion, accumulation of unresorbed primary spongiosa, was found at the metaphyses of long bones and at the costochondral junctions in Mior/Mior mice from 10 days to 37 days of age, whereas no accumulation was seen at the mid-diaphyses in these bones. The osteopetrotic conditions of Mior/Mior mice increased progressively during the first 5 weeks after birth. However, adult Mior/Mior mice 3 months or older showed improvement of the osteopetrotic condition, although the disease was not completely resolved. Ultrastructurally, osteoclasts of Mior/Mior mice had well developed ruffled borders. These results show that the Mior mutation has milder osteopetrotic changes than the original mi mutation, a surprising observation given that both mutations affect the same functional domain of the mi protein, a basic-Helix-Loop-Helix-Zipper transcription factor. The Mior phenotype resembles the intermediate autosomal recessive osteopetrosis in humans. PMID- 7495309 TI - Post-transplantation lymphoproliferative disorders arising in solid organ transplant recipients are usually of recipient origin. AB - Recent clinical, pathological, and molecular studies have increased our understanding of posttransplantation lymphoproliferative disorders (PT-LPDs). Studies have shown that the majority of PT-LPDs arising in bone marrow transplant recipients are of donor origin; however, the source (host or donor) of the lymphoid cells that make up PT-LPDs arising in solid organ transplant recipients has not been systemically investigated. In this study, 18 PT-LPDs occurring in 16 organ transplant recipients (13 heart, 2 kidney, 1 lung), 9 donor tissues (for 10 recipients), and 14 uninvolved recipient tissues (from 12 patients) were examined employing restriction fragment length polymorphism analysis to determine their host or donor origin. The PstI-digested DNAs were analyzed by Southern blot hybridization using two highly informative polymorphic probes that map to chromosome 21 (CRI-PAT-pL427-4) and chromosome 7 (CRI-PAT-pS194). All solid organ PT-LPDs with corresponding uninvolved recipient DNA showed identical hybridization patterns; none of the PT-LPDs exhibited a hybridization pattern that matched donor DNA. These findings suggest that the vast majority of PT-LPDs arising in solid organ transplant recipients, in contrast to those arising in bone marrow transplant recipients, are of recipient origin. PMID- 7495311 TI - Hypertension and peripheral arterial hemodynamics. AB - Sixty uncomplicated hypertensive patients (30 stable and 30 borderline) were studied, by strain gauge plethysmography, in comparison with 25 normotensive subjects, in order to evaluate the arterial hemodynamics of the lower limbs in essential hypertension and to verify the different pattern in borderline and in stable hypertensives. Resting blood flow, even if slightly decreased in hypertensive groups, did not show significant differences in its mean values; peak flow, instead, was reduced proportionally to the severity of hypertension in all the hypertensive patients, but only in the stable hypertensives was it statistically significant. Minimal vascular resistance showed a similar behavior: it was significantly increased only in the stable hypertensives, whereas basal vascular resistance was raised in all hypertensive patients and also in the borderline group. Finally, the half-time and the total hyperemic response time, which indicate vascular reactivity, were significantly decreased in all the hypertensives. These results suggest that the stable hypertensive patients develop principally arterial structural changes, while the borderline hypertensive patients have only functional modifications, such as a reduced compliance and a hyperdynamic condition. PMID- 7495314 TI - Transcutaneous PCO2 level as an index of tissue resistance to ischemia. AB - The authors performed a retrospective study on a data base of 525 patients with peripheral arterial disease, to analyze the pathophysiologic meaning of resting transcutaneous pressure of carbon dioxide (PCO2) and of CO2 production during three minutes of local ischemia. The resting and postischemic PCO2 and its maximum increase related to rest (PCO2 production) were measured with Kontron 7640 equipment. The results show a significant increase of PCO2 production in the Fontaine stage 2A (183 patients, 4.61 mmHg, P < 0.0001), in stage 2B (194 patients, 5.22 mmHg, P < 0.0001), in the third stage (83 patients, 6.10 mmHg, P < 0.0001), and in the fourth stage (53 patients, 8.66 mmHg, P < 0.0001). Only the patients at the first stage showed an insignificant increase, perhaps because of the small number (12) in this group. The authors feel that the measurement of tcPCO2 production during local ischemic stress can be a very important parameter for evaluating peripheral arterial disease as an expression of metabolic tissue performance and, overall, of the tissue resistance to ischemia. PMID- 7495313 TI - Efficacy and issues of emergent percutaneous transluminal coronary angioplasty. Comparison of clinical results in younger and elderly patients. AB - The authors investigated the efficacy and prognosis of emergency percutaneous transluminal coronary angioplasty (PTCA) in elderly patients. A study was conducted on the early and late prognosis of the study group composed of 66 younger patients < sixty-five years in Group A and 46 elderly patients > or = seventy years in Group B who underwent reperfusion therapy, including PTCA, for acute myocardial infarction (AMI). No difference was seen in the initial success rate of 82% for Group A and 85% for Group B, or in the vessel patency in the predischarge coronary arteriogram (CAG) with 84% for Group A and 87% for Group B. However, in-hospital mortality was 3% for Group A and 11% for Group B. The patient restenosis rate in the 4.2 months follow-up CAG was 28% for Group A and 50% for Group B. The lesion restenosis rate was 30% for Group A and 53% for Group B (P < 0.05), and the late period mortality rate was 3% for Group A and 11% for Group B. Although there was no difference in the initial success rate of reperfusion through PTCA for AMI in the elderly as compared with the younger patient group, poorer results were seen in the restenosis rate and mortality rate. PMID- 7495312 TI - Treating hyperlipidemia in the elderly. AB - BACKGROUND: Determination of the effects of dietary modification and hyperlipidemic medications in the elderly (> sixty-five years of age) patient has not been significantly investigated to date despite knowledge that elevated cholesterol (TC) and triglyceride (TG) levels increase the risk of coronary artery disease (CAD). METHODS: Twenty-seven individuals were placed into one of three treatment groups and longitudinally followed up to examine the effects of diet and hyperlipidemic medications on TC and TG levels. Group 1 (n = 14) received neither dietary nor drug therapy. Group 2 (n = 9) received dietary counseling without concomitant hyperlipidemic medications. Subjects in group 3 (n = 4) underwent dietary instruction for six months and hyperlipidemic medication(s) for eighteen months. RESULTS: Subjects in group 1 demonstrated a statistical increase in TC (P < or = 0.001) during the study. Patients in groups 2 (P < or = 0.001) and 3 (P < or = 0.05) demonstrated statistical improvement in TC reduction during dietary counseling. The effect on TC was blunted in group 3 after dietary counseling was discontinued. Reductions in TG levels were significant (P < or = 0.001) only for patients in group 2. CONCLUSION: Elderly individuals were able to significantly reduce both TC and TG levels by dietary modification alone. Minimal improvement was seen with the addition of hyperlipidemic medications. PMID- 7495315 TI - Transesophageal echocardiographic evaluation of left main coronary artery. AB - Transesophageal echocardiography (TEE) demonstrated the left main coronary trunk in 30 of 32 patients. Of these 30 patients, 10 had angiographically normal and 20 stenotic left main coronary arteries. The TEE study revealed no stenosis in 9 of 10 patients with normal left main trunks and atherosclerotic lesions in all 20 patients with stenotic left main trunks. TEE demonstrated additional areas of stenosis in the left main coronary artery in 3 patients, which included ostial lesions in 2 patients. These features were not observed by angiography. Furthermore, when compared with angiography, TEE overestimated severity of stenosis in 4 patients. In conclusion, TEE is a useful test in demonstrating normal as well as stenotic left main coronary trunks. PMID- 7495316 TI - Deep venous thrombosis in Assir region of Saudi Arabia. Case-control study. AB - In a five-year case-control study (1988 to 1993) at Assir Central Hospital (ACH), Abha (8,000 feet above sea level), Saudi Arabia, 92 of 129 patients suspected of deep venous thrombosis (DVT) were studied with ascending contrast venography (CV) (74 patients, 80.4%) or Doppler ultrasonography (DUS) (18 patients, 19.6%). Female-to-male ratio was 2.3 to 1. Age range of patients was twelve to ninety years; mean age was 44.45 yrs +/- 17.38 years. DVT hospital incidence was 18 per 10,000 admissions. The most common associated factors included immobilization due to chronic diseases (21.7%), trauma and surgery (19.6%), and pregnancy and oral contraceptives usage (16.3%). The most common symptom and sign were limb pain and tenderness (95.6%). Limb swelling was noted in 93.5% of patients. The left lower limb was more commonly affected than the right. There was a definite increase of DVT during the winter months. Altitude was not a contributory factor. Pulmonary embolism was the greatest complication. PMID- 7495317 TI - Efficacy of femoral intra-arterial administration of teicoplanin in gram-positive diabetic foot infections. AB - In this study the efficacy and safety femoral intra-arterial administration of teicoplanin in the treatment of diabetic foot infections caused by gram-positive bacteria were evaluated. Twenty-five hospitalized diabetic patients with foot ulcers or with foot ulcers and metatarsophalangeal osteomyelitis were included in the study. In the ulcers Staphylococcus aureus was present alone in 16 patients and was associated with Pseudomonas aeruginosa in 2 patients, with Candida albicans in 2, and with coagulase-negative Staphylococcus in 1 patient. In 4 patients other gram-positive bacteria were isolated. All isolated strains were resistant to various antibiotics tested. Teicoplanin, 200 mg, was administered once a day by femoral intra-arterial injection for an average period of 14.72 +/- 7.16 days (range ten to thirty-six days). Six patients were treated with an additional antibiotic intramuscularly or intravenously because of a mixed infection. At the end of the therapy microbiological assessment confirmed that gram-positive infection was eliminated in all patients. Clinical outcome demonstrated that healing occurred in 18 patients (72%) and improvement in 7 patients (28%). No adverse drug reactions were observed during the treatment. The results demonstrate that femoral intra-arterial administration of teicoplanin was highly effective in skin- and bone-infected lesions in the diabetic foot. This method may represent a further advantage in management of this severe diabetic complication. PMID- 7495318 TI - Hydrogen sulfide poisoning: management and complications. AB - Hydrogen sulfide poisoning is a rare but devastating injury. Four of 8 cases of toxic inhalation of hydrogen sulfide are presented with special attention to the life-threatening complications. Four patients died at the scene of the accident and the remaining 4 had been hospitalized in the Intensive Care Unit (ICU). One patient died in the hospital from cardiogenic pulmonary edema and circulatory failure (pulmonary wedge pressure > 30 mmHg) and 3 patients left the hospital in good health although they presented serious complications. One of them left the hospital with a temporary pacemaker, and 1 of the 2 others died after two months owing to myocardial infarction. The authors discuss in detail the clinical features of this condition and emphasize the need for urgent and appropriate therapy as well as the need for safe environmental conditions. PMID- 7495319 TI - Isolated cortical venous thrombosis--discrepancy between clinical features and neuroradiologic findings. A case report. AB - Isolated cortical venous thrombosis (CVT) is rare, and the discrepancy between clinical features and neuroradiologic findings has not been adequately emphasized. A thirty-one-year-old woman presented with focal seizures and occasionally secondary generalization. There was no focal neurologic deficit except for Todd's paralysis. Electroencephalography showed intermittent theta waves at the left frontoparietal area. Brain computed tomography (CT) and magnetic resonance imaging (MRI) disclosed a large lesion in the left frontoparietal area. An extensive occlusion of the superior cerebral veins of the left hemisphere with a patency of the dural sinuses was noted in the cerebral angiography. Eight months later, the CT hypodense lesion disappeared. Her neurologic status was stationary, but the seizures, mostly focal, still occurred occasionally. In this report, the authors emphasize the dissociation between minor clinical features and larger CT and MRI lesions in isolated CVT. PMID- 7495320 TI - Interferon-induced coronary vasospasm. A case history. PMID- 7495321 TI - Ischemic stroke following an intramuscular injection of diclofenac. Case report. AB - Nonsteroidal antiinflammatory drugs (NSAIDs) have arterial vasospastic properties. The authors report a stroke that occurred within minutes following an intramuscular injection of diclofenac. A sixty-one-year-old man, a heavy smoker, was admitted for an acute onset of mild hemiparesis thirty minutes after a single intramuscular injection of diclofenac. The neurologic deficit was resolved at one month of follow-up. The authors suggest a temporal causal relationship between the cerebrovascular event and the injection of diclofenac. The mechanism of stroke might be related to a synergistic vasospastic property shared by the use of an NSAID and heavy smoking. PMID- 7495326 TI - Palatability vs. money: experimental study of a conflict of motivations. AB - The pleasure of eating palatable food was pitted against the displeasure of giving money away in a situation where subjects had to spend money to buy their food. Ten healthy subjects taken individually had lunch in the laboratory on four different days. During the first session, they rated the palatability of small sandwiches, of ten different varieties. In the three following sessions, they were asked to eat the same number of sandwiches as in the first session and they had to buy each sandwich at a price that increased with palatability. The rate of the price increase was varied in the three sessions. As this rate increased, the subjects moved their preference to the less palatable sandwiches. The subjects' actual behavior was predictable from the quantitative relationship of pleasure ratings and price. This result supports the hypothesis according to which behavior tends to maximize multidimensional pleasure experience. PMID- 7495325 TI - Association of body mass with dietary restraint and disinhibition. AB - The relationship of disinhibition and dietary restraint with body mass was studied in a sample of 293 women. Results suggested that higher body mass was associated with an interaction of disinhibition and dietary restraint. The association of disinhibition with higher body mass was moderated by increased dietary restraint. Symptoms of an eating disorder were more strongly associated with disinhibition than with dietary restraint. These results suggest that dieting may moderate the increased body mass associated with overeating. Psychological and eating problems associated with dietary restraint were found to be of less significance than those associated with disinhibition. PMID- 7495323 TI - The contribution of psychological and sensory factors to food preference patterns as measured by the Food Attitudes Survey (FAS). AB - The relationship between food preference patterns and several psychological and sensory variables was assessed using the Food Attitudes Survey (FAS). Previous research with the FAS, which consists of preference ratings for a variety of common, unusual and fictitious foods, showed that it provides both reliable and valid information about individual differences in food preferences and attitudes (Frank & van der Klaauw, 1994). In the studies reported here, significant correlations were found between preferences for a variety of activities (as measured by the Activity Attitudes Survey or ACT) and liking for and willingness to try foods, It was also found that individuals who report that they are unwilling to try many foods are low in general sensation seeking, and that odor pleasantness ratings significantly correlate with liking for and willingness to try foods. No associations were found between FAS performance and general phobic tendencies, optimism/pessimism or disordered eating. Multiple regression analysis revealed that responses on the ACT, sensation seeking scale, a 20-item food and eating questionnaire and odor pleasantness judgments could account for from 41 to 65% of the variance in food likes, dislikes and willingness to try foods. It was concluded that personality and sensory factors contribute to pattern of responding on the FAS, and that FAS response patterns provide an index of both attitudes toward foods and general openness to experiences and activities. PMID- 7495324 TI - Evaluation of satiety sensations and food intake after different preloads. AB - The reproducibility of three questions, related to fullness, satiety and desire to eat, rated on an unmarked triangle was verified. In four sessions 12 volunteers ate pasta with tomato sauce (520 kcal) and were asked to rate the sensations felt. There was no difference in rating scores of the replications so the proposed questionnaire provides a stable measure of sensations related to satiety. Subsequently three satiety conditions were studied. Two foods, one rich in carbohydrate, pasta (baked macaroni) and the other in protein, polpette (meatballs), were used as loads at two calorie levels and as preload before an "ad libitum" meal. All the three questions proved useful in discriminating between the different satiety conditions. The food intake underlines the specificity of satiety: subjects, after eating a preload which previously had satiated them, ate other foods in different amounts depending on the kind of preload eaten. Food intake was significantly higher after the pasta preload, furthermore "fullness" and "satiety" ratings were significantly highest after the meatball preload, suggesting that in our experimental conditions, meatballs were more satiating than pasta. In conclusion, this study highlights the validity of using several quite different questions to study hunger and satiety, together with the actual food intake. PMID- 7495322 TI - Cor triatriatum: report of an unusual variant--triatrial heart. A case report. AB - Cor triatriatum is a rare cardiac malformation presenting with symptoms and signs of pulmonary venous obstruction. Although diagnosis of this defect has been facilitated by diagnostic procedures such as echocardiography and angiocardiography, it could be missed preoperatively. The authors report a case of cor triatriatum that is unique in having two separate atrial septal defects in each side of the fibromuscular membrane and that was undiagnosed preoperatively with transthoracic echocardiography and cardiac catheterization. PMID- 7495327 TI - Adolescent food choice criteria: role of weight and dieting status. AB - The purpose of this study was to examine the relation of weight status, dieting status and several associated variables to the criteria for everyday food choice used by adolescents. Study participants were 411 students between the ages of 11 and 18, drawn from 15 schools. The adolescents rated 20 food in terms of nine food attributes (how tasty or healthful specific foods were, whether the foods were eaten by friends, and so forth). Within-person correlation coefficients were then calculated between these ratings and actual food choices as measured by a food frequency scale. The relation of weight and dieting status, as predictors of each of these correlational indices of the importance of potential food choice criteria, was then analysed using hierarchical multiple regression. In similar fashion, the relation was examined between weight and dieting status and: evaluations of food attributes (choice criteria); dietary quality; calorie, sugar and fat intake; body image; and physical activity. For a majority of food choice criteria and other variables, there was an apparent influence of weight as an independent variable. However, when dieting status was analysed simultaneously with weight, similar and stronger effects were now seen for dieting status and the effects of weight disappeared. Although some of the differences as a function of dieting status resembled differences shown previously in relation to dietary restraint, it is noteworthy that the simpler dieting variable yielded these associations. Overall, a "psychology of dieting" seems more relevant than "psychology of being fat versus being thin". This psychology appears to involve cognitive self-regulation processes. It is thus crucial that intervention programs and research studies take into account both the dieting status and the weight status of participants. PMID- 7495328 TI - The effects of fear and hunger on food neophobia in humans. AB - We examined the effects of hunger and fear on food neophobia in humans. Subjects came to the experiment five or more hours food-deprived (high hunger) or two or less hours food-deprived (low hunger) and were assigned either to give a speech (high fear) or to listen to a speech (low fear). All subjects were then given the task of selecting for tasting one member of each of ten pairs of foods, each pair consisting of one novel and one familiar food. The number of novel foods chosen was the measure of food neophobia (with fewer choices indicative of greater neophobia). The results indicated that subjects were least neophobic in the low fear-low hunger condition and were tentatively interpreted in terms of Hull's (1943) theory of behavior. That is, it was assumed that fear and hunger summated to produce different levels of drive in the various conditions, which combined with responses of different habit strength (the tendency to approach novel stimuli and the tendency to approach familiar stimuli) to produce the results obtained. PMID- 7495330 TI - Historical and sociological perspectives on food. PMID- 7495329 TI - The effect on hunger and satiety of slowing the absorption of glucose: relationship with gastric emptying and postprandial blood glucose and insulin responses. AB - To investigate the relative potency of short-term control mechanisms for carbohydrate satiety, ten fasted, healthy male volunteers consumed a 250-ml, 30% glucose drink with and without the addition of guar gum (2%). Gastric emptying, hunger and fullness ratings and blood glucose and insulin levels were monitored over the following 3 h and energy intake was recorded from a test meal given 3.5 h after the drinks. The addition of the guar gum to the glucose drink lowered both postprandial glucose and insulin levels over the following 2 h. This was associated with a reduction in the ratings for hunger and desire to eat and an increase in ratings for fullness and satiety, but energy intake from the test meal was unchanged. There was no difference between values for the half time for gastric emptying for the two drinks. The short-term increase in satiety and decrease in hunger seen when glucose absorption was slowed with guar gum is unlikely to be explained by the reduction in postprandial glycaemia or differences in gastric emptying, and instead may implicate increased contact of the carbohydrate with receptors in the small intestine and consequent enhanced release of putative satiety peptides. PMID- 7495331 TI - Baker's yeast. Some biochemical aspects and their influence in biotransformations. AB - Baker's yeast is becoming an important reagent for organic synthesis. However, on many occasions, there are problems with the experimental reproducibility, which in general is the result of the different origins of baker's yeasts. In order to explain these differences, NAD (P)+ reduction inside intact living cells from different strains was measured. The method can select cells with better reduction power in a short period of time. PMID- 7495332 TI - Active site labeling with dansyl-glutamyl-glycyl-arginyl chloromethyl ketone demonstrates the full activity of the refolded and purified tissue-type plasminogen activator variant BM 06.022. AB - BM 06.022 is a tissue-type plasminogen activator deletion variant that is comprised of the kringle 2 and the protease domain of the native molecule. BM 06.022 is expressed as inactive inclusion bodies in E. coli and transferred into the active enzyme by an in vitro folding process. Active site labeling with dansyl-glutamyl-glycyl-arginyl chloromethyl ketone provides evidence that the purified BM 06.022 is fully active and that misfolded species are completely removed by affinity chromatography on ETI-Sepharose. The comparison of the kinetics of the inhibition of BM 06.022 with that of CHO-t-PA indicates that the active centers of both enzymes are rather similar. The further evaluation of the site of interaction of BM 06.022 and DnsEGRck by mass spectroscopy and amino acid sequence analysis revealed that the inhibitor is bound selectively to His322, which is part of the catalytic triad of this serine protease. PMID- 7495333 TI - Evanescent fluorobiosensor for the detection of polyaromatic hydrocarbon based on DNA intercalation. AB - A flow-injection analysis (FIA) system coupled with an evanescent wave (EW) biosensor employing total internal reflection of fluorescence radiation (TIRF) for the detection of polyaromatic hydrocarbon that intercalates into DNA is reported. A highly fluorescent intercalator, "ethidium bromide," has been used as the reference compound for detection. The EW biosensor was developed according to the procedure described earlier (1,2). Data on the analysis of Naphthalene, 3 methylcholanthrene, 7,12-dimethylbenz(a)anthracene, 1,2-benzanthracene, and some standard reference materials supplied by the National Institute of Standards and Technology are reported. The relative ability of the polyaromatic hydrocarbon to displace ethidium bromide, based on the relative binding ratio, is found to be on the order of 7,12-dimethylbenz[a]anthracene > 3-methylcholanthrene > 1,2 benzanthracene > napthalene. PMID- 7495335 TI - Does it take sugar? A clinical role for measuring the glycosylation of IgG? PMID- 7495336 TI - Instruments to assess osteoarthritis--current status and future needs. PMID- 7495337 TI - Why the gender difference in susceptibility to rheumatoid arthritis? PMID- 7495334 TI - High-alkaline protease from Bacillus PB92 entrapped in calcium alginate gel. Physicochemical and microscopic studies. AB - High-alkaline protease (HAP) has been entrapped in Manugel DMB (an alginate gel) and assayed with two sizes and types of substrates: neutral protein casein and synthetic chromogenic tripeptide substrate, Z-Gly-Pro-Cit-PNA. Increasing the concentration of calcium chloride used for capsule formation decreased the measured enzyme activity with both substrates. Capsules were found to be stable in water for long periods of time, but they dissolved in both phosphate and carbonate-bicarbonate buffers. The pH vs activity profiles of encapsulated enzyme showed pH optima between 10 and 11 with both substrates. The calcium alginate matrix surrounding the enzyme was quite effective in stabilizing the enzyme at 20 25 degrees C and even more so at 4 degrees C. Enzyme stability at 50 degrees C was quite impressive, some enzyme activity being evident even after remaining for 1 wk at this temperature in water. Increasing concentrations of sodium dodecyl sulfate (SDS) were also found to inhibit the protease progressively, whereas a polyhexamethylene biguanidium chloride (PHMBH+Cl-) and SDS:PHMBH+Cl- combination showed the opposite effect. Optical microscopy, especially polarized light microscopy, provided a sensitive physical means of ascertaining some of the structural properties (sphericity, disorganization or organization, distinct layer enveloping the capsules, intensity of the maltese cross) of the capsules with and without enzyme before and after different chemical treatments and the presence or absence of the substrate. PMID- 7495338 TI - Is rheumatoid arthritis a failure of B cell death in synovium? PMID- 7495340 TI - Survival and drug discontinuation analyses in a large cohort of methotrexate treated rheumatoid arthritis patients. AB - OBJECTIVES: To determine the probability of drug continuation in a large cohort of methotrexate treated rheumatoid arthritis (RA) patients, the reasons for discontinuation of methotrexate, the overall survival of the members of this cohort, and the causes of death in these patients. METHODS: Yearly follow up was conducted in methotrexate treated RA patients who formed a cohort between 1981 and 1986 at a tertiary care centre. The probability of drug continuation and the patients' survival were calculated using standard statistical procedures; standardised mortality ratios were calculated using death certificate data and USA general population and mortality tables. RESULTS: The probability of methotrexate continuation at 10 years from the time the first members entered the cohort was 30%. Toxicity (and its severity) was the most frequent cause of discontinuing methotrexate. The cumulative probability of survival was 85% for women and 45% for men. A greater than expected number of deaths from infections was observed, but the number of deaths from cancer and cardiovascular diseases were within the range expected. CONCLUSIONS: Toxicity remains the most common cause for methotrexate discontinuation. Survival was comparable to that of other RA cohorts. Methotrexate may be implicated as an associated factor in the deaths from infections. PMID- 7495339 TI - Teenager with an irritable hip, anaemia and malaise. AB - We have described a boy with a juvenile spondyloarthropathy who also showed inflammatory changes in the gut. Gastrointestinal symptoms initially indicated the presence of an inflammatory bowel disease, but this was excluded histopathologically. PMID- 7495342 TI - Association of locomotor complaints and disability in the Rotterdam study. AB - OBJECTIVE: To determine the association between joint complaints and locomotor disability. METHODS: During a home interview survey 1901 men and 3135 women aged 55 years and over (the Rotterdam Study) were asked about joint pain and morning stiffness in the past month, and locomotor disability was assessed by six questions from the Health Assessment Questionnaire (HAQ). RESULTS: The prevalence of locomotor disability was 24.5% for men and 40.5% for women. The prevalence of joint pain in men was 0.7% for pain in the hips, knees, and feet simultaneously, 3.7% for pain at two joint sites, 16.0% for pain at one joint site, and 20.4% for pain in the hips and/or knees and/or feet (any joint site); the corresponding estimates for women were 1.9%, 9.0%, 23.7%, and 34.5%, respectively. The prevalence of generalised morning stiffness was 4.9% for men and 10.4% for women. The age adjusted odds ratios for locomotor disability in men ranged from 2.4 of pain at one joint site to 8.8 of pain at all three joint sites; for women these odds ratios varied between 2.5 and 5.7, respectively. The age adjusted odds ratios of generalised morning stiffness were 8.0 for men and 7.3 for women. CONCLUSION: There is a strong and independent association between locomotor disability and age, joint pain, and generalised morning stiffness in people aged 55 years and over. The odds for locomotor disability increase onefold for every year increase in age, while the presence of generalised morning stiffness is of greater influence than the presence of joint pain. PMID- 7495343 TI - Lp(a) lipoprotein levels as a predictor of risk for thrombogenic events in patients with Behcet's disease. AB - OBJECTIVES: To investigate whether plasma levels of Lp(a) lipoprotein (Lp(a)) are predictors of defective fibrinolytic activity, leading to thrombosis, in patients with Behcet's disease. METHODS: Plasma Lp(a) was measured by enzyme linked immunosorbent assay, lipids and lipoproteins by enzymatic methods, and apolipoproteins A-I and B, fibrinogen (turbidimetric method), tissue plasminogen activator (t-PA), plasminogen activator inhibitor-1 (PAI-1), and D-dimer levels by enzyme immunoassay. Their levels and interactions were evaluated in 33 patients with Behcet's disease (including five with thrombotic complications) and 30 healthy control subjects. RESULTS: Plasma Lp(a) concentration was significantly greater in the patients than in controls. Nine patients (27%) had levels > 0.30 microgram. There was no correlation between Lp(a) and other lipids and lipoproteins apart from apolipoprotein B. Lp(a) showed inverse correlation with t-PA (r = -0.34, p < 0.05) and D-dimer (r = 0.35, p < 0.05). Patients with thrombotic complications had significantly greater Lp(a) and PAI-1, and lower D dimer concentrations than control subjects. CONCLUSIONS: Measurement of plasma Lp(a) levels in patients with Behcet's disease may provide useful information regarding the potential development of thrombotic events, because of its possible role in defective fibrinolysis. PMID- 7495345 TI - Rheumatology education and management skills in general practice: a national study of trainees. AB - OBJECTIVE: To evaluate current rheumatology education and skills during vocational training in general practice. METHODS: A postal questionnaire survey was sent to all general practice trainees who were in United Kingdom training practices in June 1992. A second survey was made of general practice trainers in the UK. RESULTS: Questionnaires were returned by 1624 trainees, representing 70% of all trainees known to be in a training practice on 1 April 1992. Of the 1075 responders who were at the end of their trainee year, 35% had not received any tutorials on rheumatological topics with their trainer, and only 43% had experienced specific rheumatology teaching on local day release courses. Although 84% of these trainees had injected or aspirated the knee, fewer than 40% had acquired shoulder injection skills. Lack of experience was matched by low reported confidence. Trainees rated the amount of their rheumatology education as inadequate. Nine hundred trainers returned questionnaires--a response rate of 33%. Their estimate of the amount of local training provided was similar to that among the trainees, but this sample of trainers reported a higher level of practical teaching than the trainees indicated they had received. CONCLUSIONS: Rheumatology education during vocational training needs to be improved, particularly the component provided by trainers and local day release courses. This process might be facilitated by the development of a standard rheumatology curriculum which could be incorporated into all training schemes. PMID- 7495344 TI - Plasma endothelin-1 levels in patients with systemic sclerosis: influence of pulmonary or systemic arterial hypertension. AB - OBJECTIVES: To investigate the behaviour of circulating endothelin-1 (ET-1) in patients affected by systemic sclerosis and to elucidate the relationship between systemic and pulmonary plasma peptide and arterial pressure levels. METHODS: Plasma ET-1 concentrations were determined in 48 patients affected by systemic sclerosis (41 women, seven men; mean age 47.2 (SD 5.5) years) with or without systemic or pulmonary hypertension (or both). A group of 18 normal volunteers served as controls (15 women, three men; mean age 45.0 (10.1) years). RESULTS: Plasma ET-1 levels were significantly greater in patients affected by systemic sclerosis (1.65 (0.29) pg/ml) than in controls (0.63 (0.19) pg/ml) (p < 0.0001). Pulmonary artery systolic hypertension alone was present in 14 patients with systemic sclerosis (50.5 (8.49) mm Hg, range 37-67 mm Hg), and systemic hypertension alone (160.7 (5.9)/100.6 (3.2) mm Hg) was present in 11 patients. Both conditions were present in 12 patients, while 11 patients had systemic hypertension. There were no significant differences in plasma ET-1 levels between patients with pulmonary hypertension alone (1.62 (0.21) pg/ml) and those with systemic hypertension alone (1.65 (0.43) pg/ml). In particular, patients with normal pulmonary artery and systemic pressures (n = 11) had plasma ET-1 concentrations identical to those found in patients (n = 12) with both pulmonary and systemic hypertension (1.70 (0.15) v 1.64 (0.35) pg/ml, respectively). No correlations were observed between plasma ET-1 and either pulmonary or systemic pressures. CONCLUSION: Systemic sclerosis is characterised by increased plasma ET 1 levels, but neither pulmonary nor systemic hypertension are accompanied by further increase in plasma peptide levels. PMID- 7495341 TI - Increased levels of serum IgM antibody to staphylococcal enterotoxin B in patients with rheumatoid arthritis. AB - OBJECTIVE: To investigate the role of superantigen in rheumatoid arthritis (RA) by assaying the serum levels of staphylococcal enterotoxin B (SEB) antibodies. METHODS: Serum IgG and IgM SEB antibodies were measured using an enzyme linked immunosorbent assay (ELISA), and confirmed by Western blot analysis. The T cell receptor V beta (TCR V beta) repertoire was analysed using the reverse transcriptase polymerase chain reaction. RESULTS: RA patients had increased levels of serum IgM SEB antibody compared with normal subjects, patients with systemic lupus erythematosus, Sjogren's syndrome, and Behcet's disease. The titres of rheumatoid factor (RF) showed no correlation with the levels of IgM SEB antibodies, and the levels of SEB antibodies were not inhibited by the addition of human immunoglobulin, or after absorption of RF. RA patients whose disease duration was less than 10 years had greater levels of serum IgM SEB antibodies than those with disease duration more than 10 years. The levels of IgM and IgG SEB antibodies in synovial fluid from RA patients were correlated with those in their sera. Western blot analysis detected IgM and IgG SEB antibodies as a band of approximately 30 kDa molecular size. The percentage of TCR V beta 2, V beta 5.2, and V beta 12 in phytohaemagglutinin stimulated peripheral T cells correlated significantly with the levels of serum IgM SEB antibody in RA patients. CONCLUSION: These results suggest that SEB, one of the superantigens, may have a critical role in the pathogenesis of RA. PMID- 7495346 TI - Rheumatic fever: autoantibodies against a variety of cardiac, nuclear, and streptococcal antigens. AB - OBJECTIVE: To measure antibody titres to cardiac, nuclear, and streptococcal antigens in different groups of rheumatic fever (n = 60) and control subjects (n = 80) with the aim of identifying cross reactive antigens of potential laboratory diagnostic value. METHODS: Enzyme linked immunosorbent assays (ELISA), immunocytochemical, and electrophoretic techniques were used to measure titres of antibodies to a variety of cardiac, nuclear, and streptococcal antigens in seven groups comprising patients with rheumatic fever and control subjects. RESULTS: Increased concentrations of antibodies to several streptococcal and cardiac antigens, in addition to increased IgA and IgG levels, were noted in sera from patients with acute rheumatic fever and chronic rheumatic heart disease. Autoantibodies to nuclear antigens were evident in three rheumatic fever sera. CONCLUSION: Although we were unable to identify any unique cross reactivity between cardiac and streptococcal antigens, these results demonstrate that there is an exaggerated humoral response to several cardiac, nuclear and streptococcal antigens in patients with rheumatic fever. PMID- 7495347 TI - Transforming growth factor beta 2 in labial salivary glands in Sjogren's syndrome. AB - OBJECTIVE: To compare the distribution and the amount of transforming growth factor beta (TGF beta) in labial salivary glands (LSG) in patients with Sjogren's syndrome (SS) and healthy controls. METHODS: LSG from SS patients (n = 10) and healthy controls (n = 6) were labelled with peroxidase-antiperoxidase staining for TGF beta 2, which was quantitated in image analysis using Video Interactive Display System software. RESULTS: In all LSGs in SS and healthy controls, TGF beta 2 was found in endothelial cells of the capillaries and in the capsular and stromal fibroblasts. In LSGs in SS, TGF beta 2 was also found in some lymphocytes in the inflammatory cell foci and in fibroblasts in fibrotic areas. The TGF beta 2 staining index (microns 2/mm2 tissue) was greater in SS than in control LSGs (3670 (SEM 430) v 2061 (176); p < 0.01), with no difference between the primary and secondary forms of SS (p > 0.05). CONCLUSION: The localisation and the level of expression of TGF beta 2 indicate its involvement in local tissue fibrosis, and may reflect attempts at immunosuppression. PMID- 7495348 TI - Primary Sjogren's syndrome in men. AB - OBJECTIVE: To describe the clinical expression of primary Sjogren's syndrome (SS) in men, focusing on extraglandular manifestations (EGM) and serological markers of disease. METHODS: In a cross sectional and comparative study, adult men with primary SS were identified from a cohort study on SS, and 26 age matched adult women with primary SS were selected as a control group. All patients met the European classification criteria for SS. They were compared for demographic, clinical and laboratory findings. RESULTS: Thirteen men with primary SS were identified. Mean age at onset was 39 (SEM 4) years and mean duration of disease was 7.8 (1) years. Sicca complex or parotitis was the presenting feature in eight patients (61.5%), and an EGM in five (38.5%). During the course of the disease, EGM were present in 12 patients (92%), polyarthralgias and lymphopenia being the most frequent (38.5% each). Rheumatoid factor was positive in 73% of patients, antinuclear antibodies in 85%, anti-(SS-A) in 62%, and anti-(SS-B) in 46%. No statistical differences in the frequency of EGM or in the presence of autoantibodies were observed between men and women. However, men patients were more likely to have EGM. CONCLUSION: Primary SS in men is an uncommon condition with clinical and serological characteristics similar to those observed in women. Sex hormones may be incriminated in the pathogenesis of SS. However, it remains poorly understood whether sex hormones play a major role in the severity of disease and have any importance with regard to treatment. PMID- 7495349 TI - Increased serum IgE level and interleukin-4 release from cultured lymphocytes from a patient with adult onset Still's disease. AB - OBJECTIVE: To evaluate the relationship between high serum levels of IgE and the release of interleukin-4 (IL-4) from cultured lymphocytes of a patient with adult onset Still's disease. METHODS: IL-4 concentrations in plasma and culture from inactivated peripheral blood mononuclear cells were assessed by enzyme immunoassay during febrile episodes and remission. RESULTS: A high level of IL-4 was detected by enzyme immunoassay in the peripheral blood mononuclear cells cultured from the patient. These seemed to correspond with a febrile episode and a high serum IgE concentration. CONCLUSION: Increased serum IgE concentrations during a febrile episode are rare in patients with adult onset Still's disease, but the relationship between the high levels of serum IgE and IL-4 in cultured lymphocytes may provide clues to pathogenesis of the condition. PMID- 7495350 TI - Maternal age influences risk for HLA-B27 associated ankylosing enthesopathy in transgenic mice. AB - OBJECTIVE: To study further the temporal clustering of ankylosing enthesopathy (AE) noted originally during a study of the influence of mouse major histocompatibility complex (MHC) H-2 and transgenic HLA-B27 on the frequency of AE. METHODS: The relationship between maternal age at littering and frequency of AE was analysed. RESULTS: Mice born to mothers aged eight months or older had a significantly lower disease frequency of AE than mice born to mothers younger than eight months of age. This phenomenon was observed in three independent cohorts evaluated to date (p < 0.01, 0.025, and 0.05). CONCLUSION: Maternal age is a novel, non-genetic risk factor as defined in relation to an MHC associated enthesopathy. Its mode of action and relevance to human disease require further investigation. PMID- 7495352 TI - Cytoplasmic staining, ANA negative status, and ENA testing in rheumatic diseases. PMID- 7495351 TI - 3rd International Symposium on the Immunotherapy of the Rheumatic Diseases. 10-14 May 1995, Cyprus. Abstracts. PMID- 7495353 TI - Effect of gold treatment on cytokine expression in synovium. PMID- 7495354 TI - Relapsing polychondritis and Behcet's syndrome in a patient with HIV infection. PMID- 7495355 TI - Human leucocyte antigen typing in rheumatoid arthritis/polymyositis overlap syndrome. PMID- 7495356 TI - Cytidine deaminase is not released from degranulating polymorphonuclear neutrophils. PMID- 7495357 TI - Carpal tunnel syndrome as initial manifestation of inflammatory connective tissue diseases. PMID- 7495358 TI - Human papillomaviruses, cervical cancer and the developing world. AB - Carcinoma of the uterine cervix is the commonest cancer of women in the majority of developing countries; in developed countries it is much less common, as well as being less frequent than some other cancers, such as those of breast, lung and colon. The discovery of human papillomavirus (HPV) as an aetiological agent of cervical cancer (and of other anogenital malignancies) has prompted a great deal of interest in the biology and oncogenicity of this virus, including large-scale epidemiological surveys comparing the prevalence of HPV and other possible causative agents between areas of high and low incidence of cervical carcinoma. These studies have thrown up tantalizing clues concerning the virus' pathogenesis and disease associations. Sexual practices, parity, diet, smoking, other diseases (including sexually transmissible diseases) and immunogenetic characteristics may all play a contributory role in the development of cervical carcinoma. These factors may be independent of each other and of association with HPV infection. Immunization to prevent papillomavirus infection and thus cervical cancer is an attractive prospect. Although the potential in using such an approach has been demonstrated in several animal models, there are several major theoretical problems to solve before HPV vaccination becomes a practicality. Intervention to prevent exposure to other aetiological factors and to improve screening may be a more practicable strategy for reducing the burden of cervical cancer. PMID- 7495359 TI - The chemotherapy of rodent malaria. LII. Response of Plasmodium yoelii ssp. NS to mefloquine and its enantiomers. AB - A comparison was made between the blood schizontocidal action in mice of racemic mefloquine hydrochloride and the free bases of its (+)- and (-)-enantiomers (Ro 13-7224 and Ro 13-7225) against chloroquine-resistant Plasmodium yoelii ssp. NS. The racemic hydrochloride was two to three times as active against this parasite in mice as either of the enantiomer free bases, which were of similar activity to each other. Under drug selection pressure, the parasites acquired resistance in approximately the same time for each of the three compounds. PMID- 7495360 TI - Efficacy and tolerability of a sequential, artesunate suppository plus mefloquine, treatment of severe falciparum malaria. AB - Thirty patients with severe falciparum malaria were each given a total of 1600-mg artesunate suppository over three consecutive days followed by 1250 mg mefloquine per os, divided into two doses which were given 12 h apart. All patients were admitted for 28 days to the Bangkok Hospital for Tropical Diseases, so that the efficacy and tolerability of the treatment could be assessed. All the patients showed clinical improvement, with mean (S.D.) parasite and fever clearance times of 50.4 (13.0) and 70.7 (44.9) h, respectively. Two patients with unrousable coma (Glasgow coma score < or = 8) on admittance regained consciousness 46 and 48 h post-treatment. One other patient had acute renal failure and required dialysis. Most patients (80%) were initially hyperparasitaemic, with a mean density of 184,344 parasites/microliters blood. No deaths occurred. Efficacy was evaluated in 25 of the patients. The cure rate 28 days post-treatment was 92%. None of the patients had major adverse effects although two had tenesmus and passed stools immediately after each suppository was administered. A fresh suppository had to be inserted when this occurred. The results indicate that artesunate suppositories followed by oral mefloquine constitute a well-tolerated regimen with a high cure rate. The combination is suitable as an alternative treatment for severe malaria, particularly in children. Further, large-scale studies are required. PMID- 7495361 TI - Evidence from genotypic and phenotypic markers that an attenuated line outgrows a virulent one in a mixed population of Leishmania major promastigotes cultured in vitro. AB - Two cloned lines of Leishmania major promastigotes, one attenuated (CO1H) and one virulent (CO1R), differing in molecular karyotype and expression of the major surface glycoprotein (gp63), were mixed to produce two heterogeneous populations: MP-1 (100 CO1R promastigotes/CO1H promastigote); and MP-2 (10,000 CO1R promastigotes/CO1H promastigote). The mixed populations were cultured for 1 month in vitro in HO-MEM medium and sub-samples taken on days 4 and 30 were subjected to electrophoresis so that the molecular karyotypes and gp63-expression characteristics of the promastigote populations could be determined. In spite of the initial predominance of the virulent CO1R, the attenuated CO1H always outgrew it. The patterns of growth of pure cultures of CO1H or CO1R did not fully explain this observation. When grown alone, CO1H acidified the culture medium much more and much faster than CO1R, low pH values eventually inhibiting multiplication. Decreasing the medium's initial glucose concentration or increasing its initial pH prolonged the growth phase of CO1H, probably by slowing its acidification of the medium to inhibitory values. It is possible that, in mixed populations, the CO1R promastigotes help to buffer the medium, permitting faster and longer multiplication of the CO1H promastigotes than occurs when they are cultured alone. CO1H promastigotes may also inhibit CO1R multiplication; CO1H promastigotes in mid-logarithmic phase entered stationary phase within a day of being transferred into cell-free supernatants from 3-day-old CO1R cultures. PMID- 7495362 TI - Immunoblot evaluation of IgG and IgG-subclass antibody responses for immunodiagnosis of human alveolar echinococcosis. AB - Antigen binding of total-IgG and IgG-subclass antibodies from patients with alveolar or cystic echinococcosis (AE and CE) was assessed by immunoblotting. Antigen extracts were prepared from Echinococcus multilocularis protoscoleces (EmP) or from homogenized E. multilocularis metacestode tissue (EmCH). Antigens of approximately 44, 35, 21, 17.5 and 16.5 were recognized by total-IgG and IgG1- and IgG4-subclass antibodies in some of 50 human AE sera from China, Japan or France. The 44- and 35-kDa polypeptides, present in both EmP and EmCH extracts, were recognized by total-IgG antibodies in sera from 82% and 66% of the AE patients, respectively. However, over 30% cross-reactivity occurred between these two antigens and sera from CE and Taenia solium cysticercosis patients. The immunoblot specificities of the 27-, 21- and 17.5-kDa antigens in EmP for E. multilocularis infection were 73%, 88% and 93%, respectively. Recognition of the 17.5-kDa antigen in the EmP immunoblot was much higher for the Japanese AE cases (11/13; 85%) than for the French (9/19; 47%) or Chinese (9/18; 50%) AE cases. None of the CE cases from Uruguay or Libya, where human AE has not been reported, was seropositive for the 17.5-kDa antigen. Antibodies from three (7.3%) of the 41 Chinese CE cases recognized the 17.5-kDa antigen. Within the 13 Japanese AE sera, the combined detection by IgG1, IgG4 and total-IgG antibodies of the 27-, 21- and 17.5-kDa antigens in either EmP or EmCH immunoblots was greater than that by each class/subclass alone, increasing the overall sensitivity for AE patients. A combined ELISA/immunoblot approach, including IgG-subclass detection using E. multilocularis protocolex or cyst extracts, could be useful for the differential diagnosis of human alveolar echinococcosis. An algorithm for such an approach is given. PMID- 7495363 TI - Experimental murine schistosomiasis: reduced hepatic morbidity after pre- and/or post-infection treatment with ibuprofen or diclofenac sodium. AB - Arachidonic-acid metabolites appear to participate in skin penetration by and transformation of schistosome cercariae and in the pathogenesis of schistosomiasis. With this in mind, mice were treated with one of two cyclooxygenase inhibitors before and/or after infection with Schistosoma mansoni. The effects of the treatment on liver morbidity and the parasitic infection were then evaluated, using infected, untreated and uninfected, treated mice as controls. Treatment with ibuprofen (20 mg/kg.day) or diclofenac sodium (2.5 mg/kg.day) for 7 days before infection led to significantly lower liver weights, worm loads and hepatic hydroxyproline contents than in the untreated mice. If treatment with either drug was continued after infection, for 28 days, there was an additional significant decrease in hepatic gamma-glutamyl transpeptidase activity. All these parameters except liver weight were similarly affected when treatment with either drug was begun on the day of infection and continued for 28 days. There was no significant change in liver weight or worm load when treatment was delayed until day 28 post-infection but faecal egg counts were reduced in the treated groups. In additional experiments, using a smaller dose of diclofenac sodium (1.25 mg/kg.day), all the measured parameters of infection were significantly decreased when the treatment was initiated 7 days before infection and continued until day 28 post-infection. The results indicate that the treatment of S. mansoni-infected mice with ibuprofen or diclofenac sodium was effective in reducing the severity of infection and in attenuating hepatic fibrosis, particularly when the treatment was started early in relation to the time of infection. PMID- 7495364 TI - Ascaris lumbricoides infection and environmental risk factors in an urban African setting. AB - Identification of appropriate strategies for controlling gastro-intestinal nematodes in communities depends, in part, on an understanding of the conditions that increase risk of exposure to infective stages. The present study was conducted in Lubumbashi, Zaire. The objectives were to identify features of the environment and living conditions that were significant predictors of Ascaris lumbricoides infection, and to determine whether the same predictors were important in populations living in subdivisions of lower (LSES) and higher (HSES) socio-economic status. Forty-two households from each of three subdivisions (two LSES and one HSES) were selected at random. Mothers were interviewed, observations on the environment around the home were recorded, and single stool samples, collected from all children and mothers, were examined for nematode eggs. Maternal education was a significant predictor of A. lumbricoides intensity in both LSES and HSES subdivisions. Factors related to poor sanitation (open defecation and high number of people using the same toilet) were important in the LSES subdivisions but not in the HSES subdivision. In contrast, the ratio of relatives to direct family members per household was a significant predictor of high intensity of infection in the HSES subdivision, but not in the LSES subdivisions, indicating that relatives and live-in visitors contribute to Ascaris transmission in the HSES population. PMID- 7495365 TI - The prevalence and clinical features of multi-drug resistant Salmonella typhi infections in Baluchistan, Pakistan. AB - Between January and July 1994, a prospective study of bacteraemia in 692 patients with fever without localizing signs was undertaken at the Quetta Military Hospital in Baluchistan, Pakistan. Salmonella spp. were isolated from 76 (11%) of the patients; 62 had S. typhi and 14 had S. paratyphi A. Significantly more isolations of S. typhi were made in the hot dry months of May and June than in the earlier months. Although multi-drug resistance (to chloramphenicol ampicillin and cotrimoxazole) was detected in 43 (69%) of the S. typhi isolates, it was not found in any of the S. paratyphi A. Defervescence of patients with chloramphenicol-sensitive S. typhi took 7-10 days of chloramphenicol therapy. In contrast, most (91%) of the patients infected with multi-drug resistant S. typhi who were treated with fluoroquinolones achieved defervescence in 1-3 days; the remainder took 4-6 days. PMID- 7495366 TI - Comparative effects of permethrin-impregnated bednets and DDT house spraying on survival rates and oviposition interval of Anopheles farauti No. 1 (Diptera:Culicidae) in Solomon Islands. AB - Human-biting, CDC light trap and pig-baited collections were used to monitor changes in the abundance and parity rate of Anopheles farauti No. 1 mosquitoes in three Solomon Islands villages for 30 consecutive nights. Houses in one of the villages were sprayed with DDT, another village was unsprayed but its inhabitants used permethrin-treated bednets and the third village was left completely untreated. Mosquitoes collected each day were dissected to determine follicular maturation and parity. In the second year of vector control, survival rates (determined by time-series analysis of the landing catches and biting parous population) were significantly lower in the village using permethrin-treated bednets than in the other two villages. There was no difference in the estimated survival rates between the untreated and DDT-sprayed villages. The oviposition cycle was extended to 4 days in the permethrin-treated village compared with 3 days in the other villages. The expected infective life was 1.75-fold longer in the untreated village than in the permethrin-treated village. The epidemiological implications of replacing DDT spraying with permethrin for malaria control are discussed. PMID- 7495367 TI - Blood-digestion kinetics of four Anopheles species from Trinidad, West Indies. AB - The blood-digestion kinetics of Anopheles albitarsis, An. aquasalis, An. bellator and An. homunculus were determined in the laboratory using females collected from two field sites in Trinidad. Anopheles aquasalis displayed the highest rate of haemolysis (giving an absorbance of 0.36 at 410 nm), followed by An. albitarsis (0.16), An. bellator (0.07) and An. homunculus (0.05). Trypsin activity peaked 12 24 h after blood feeding and then declined to zero at 60 h in all four species. Anopheles albitarsis had significantly higher maximum trypsin activity (0.69 units) than An. aquasalis (0.28), An. bellator (0.18) or An. homunculus (0.12) (P < 0.01). Aminopeptidase activity patterns were similar for An. aquasalis, An. bellator and An. homunculus, with peak activity at 18 h. Among the An. albitarsis mosquitoes, peak aminopeptidase activity occurred at 24 h. The peritrophic membrane developed 18, 30, 30 and 36 h post-feeding in An. aquasalis, An. albitarsis, An. bellator and An. homunculus, respectively. Stage V ovarian follicles began to mature 36 h after An. albitarsis and An. bellator fed to repletion and after 48 h in An. aquasalis and An. homunculus. Ovarian development in the four species was not affected by patterns of erythrocyte haemolysis, proteolytic enzyme activity or peritrophic-membrane development. The inter- and intra-specific variations observed in the blood-processing physiology of the four species of Anopheles are briefly discussed in terms of phylogeny. PMID- 7495368 TI - Prospects and problems of intermittent irrigation for control of vector breeding in rice fields in southern India. AB - The impact of water management on the abundance and interaction of aquatic stages of mosquitoes and some important groups of their predators was studied in a 22.3 ha plot of rice in southern India. Many pools persisted even in a year when intermittent irrigation (II) was properly implemented (fields being re-irrigated as soon as they were dry) and these acted as microrefuges for the immature stages of mosquitoes and for their predators. Although the abundance of all predators, particularly of Notonectidae, was reduced by II in this year, increased predation efficiency in the confined spaces of the persistent pools resulted in a 75%-88% reduction in the abundance of mosquito pupae, as measured by two independent sampling methods. In another year there was insufficient irrigation water for proper II and re-irrigation took place whenever water was available rather than when it was required, frequently resulting in only partial drying. This reduced predator populations without enhancing their efficiency, and so mosquito abundance was slightly higher than in conventionally irrigated fields. The implications for vector control are discussed. PMID- 7495369 TI - Phlebotomine sandflies in Venezuela. IV. Review of the Lutzomyia subgenus Micropygomyia (Diptera:Psychodidae), with a description of L. absonodonta n. sp. and the male of L. lewisi. AB - The subgenus Micropygomyia, of the genus Lutzomyia, is reviewed. A new species, Lutzomyia absonodonta, previously misidentified as L. micropyga, and the male of L. lewisi are described. Illustrated keys for identifying male and female flies of the Micropygomyia species in Venezuela, distribution maps and notes on the ecology of each species are also provided. PMID- 7495370 TI - Detection of antibodies to surface antigens of Angiostrongylus cantonensis by ELISA. PMID- 7495371 TI - Apparent lack of vertical transmission of simian immunodeficiency virus (SIV) in naturally infected African green monkeys, Cercopithecus aethiops. PMID- 7495372 TI - Inhibitory effect of methylmercury on migration and tube formation by cultured human vascular endothelial cells. AB - The effect of methylmercury chloride (MeHg) on migration and tube formation by cultured human umbilical vein endothelial cells (HUVECs) was quantitatively analyzed. The distance of endothelial cell outgrowth from the scraped edge of a monolayer was measured. HUVEC outgrowth was inhibited by MeHg (1.0-5.0 microM) treatment in a dose-dependent manner. Tube formation was studied by culturing the cells on gelled basement membrane matrix (Matrigel). Treatment of HUVECs with 0.1 5.0 microM MeHg for 24 h inhibited tube formation dose-dependently. These results suggest that migration and tube formation by HUVECs are susceptible to MeHg cytotoxicity, and that MeHg could be injurious to endothelial cell function, which may be involved in the pathogenesis of arteriosclerosis. PMID- 7495373 TI - Formation of epoxide and quinone protein adducts in B6C3F1 mice treated with naphthalene, sulfate conjugate of 1,4-dihydroxynaphthalene and 1,4 naphthoquinone. AB - Naphthalene (NA) is metabolically activated to the reactive intermediates, naphthalene oxide (NO) and naphthoquinones. To investigate the role of circulating reactive metabolites in NA toxicity, the half-life of NO was examined. The in vitro half-life of NO in both whole blood and plasma was 10 min. Detectable levels of NO were seen in perfusate leaving the isolated perfused liver of B6C3F1 mice infused with 10 mumol/h NA. Identification of protein sulfhydryl adducts in NA-exposed mice (50 and 100 mg/kg, IP, 24 h) revealed a predominance of quinone adducts in liver, lung, kidney, red blood cells and brain. The epoxide adduct predominated in plasma protein. Administration of the sulfate conjugate of 1,4-dihydroxynaphthalene (NHQS) (100 mg/kg) resulted in formation of naphthoquinone protein sulfhydryl adducts in lung, liver and kidney. Administration of 1,4-naphthoquinone (NQ) (5 mg/kg) produced NQ adducts in liver, lung, kidney, plasma and brain. PMID- 7495374 TI - Interactive effects of 2,3,7,8-tetrachlorodibenzo-p-dioxin and retinoids on proliferation and differentiation in cultured human keratinocytes: quantification of cross-linked envelope formation. AB - Dioxins are potent inducers of chloracne in humans. This skin aberration can be interpreted as an altered differentiation pattern of acinar sebaceous base cells and a change in the rate of terminal differentiation of the keratinocytes. We measured this rate induced by 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD) in primary cultures of human keratinocytes. As parameters for differentiation, we quantified the 35S-methionine incorporation into cross-linked envelopes (revealing the total CLE biomass), as well as the number of microscopically visible CLEs. It was shown that TCDD is a very potent inducer of both CLE biomass and number with a half-maximal effect concentration (EC50) of 1.4 nM. CLE biomass was maximally increased 10-fold and the number of cells in culture producing a CLE was increased from 15% in control cultures to maximally 75% of the cells in TCDD-treated cultures. Both effects were Ca(2+)-dependent and increased with elevated cell density, being optimal in post-confluent cultures. Retinoic acid dose-dependently decreased the effect of 10(-8) M TCDD, 10(-6) M having a nearly complete antagonistic action. This interaction of retinoic acid with TCDD-induced differentiation was non-competitive. Retinol was equally potent as an antagonist of the TCDD-induced elevation of CLE formation as compared with retinoic acid. Retinyl palmitate and etretinate were not very effective as TCDD antagonists. Supplementation of hydrocortisone suppressed the TCDD-induced keratinocyte differentiation. It was concluded that CLE biomass quantification provides a reliable and sensitive parameter for keratinocyte differentiation. In this in vitro system it is shown that TCDD strongly induces a switch from proliferation to terminal differentiation and that this effect can be antagonized effectively by retinoic acid and retinol. PMID- 7495375 TI - Acute toxicity of cyclohexylmethylphosphonofluoridate (CMPF) in rhesus monkeys: serum biochemical and hematologic changes. AB - Changes in serum biochemical and hematological parameters were studied in 20 male rhesus monkeys following acute poisoning by the organophosphate nerve agent cyclohexylmethylphosphonofluoridate (CMPF or GF). Animals were challenged with 5 x LD50 GF (233 micrograms/kg, IM) following pretreatment with pyridostigmine (0.3 0.7 mg/kg per 24 h) and treated with atropine (0.4 mg/kg, IM) and either 2-PAM (25.7 mg/kg, IM) or H16 (37.8 mg/kg, IM) at the onset of clinical signs or at 1 min after exposure. Muscle fasciculations, tremors, or convulsions occurred in 19 of 20 animals. Serum biochemical and hematologic parameters were analyzed 2 days and 7 days after exposure and compared to pre-exposure baseline values. Significant increases in creatine kinase (CK), lactate dehydrogenase (LD), aspartate transaminase (AST), alanine transaminase (ALT) and potassium ion (K+), associated with damage to striated muscle and metabolic acidosis, occurred in both oxime-treated groups 2 days after exposure. Total protein, albumin, red blood cell (RBC) count, hemoglobin concentration (Hb) and hematocrit (Hct), were decreased in both oxime-treated groups at 7 days. The results demonstrate that animals exposed to a single high dose of GF and treated with standard therapy exhibit changes in serum biochemical and hematological indices directly and indirectly associated with their clinical presentations. PMID- 7495376 TI - Assessment of primary neuronal culture as a model for soman-induced neurotoxicity and effectiveness of memantine as a neuroprotective drug. AB - An in vitro mammalian model neuronal system to evaluate the intrinsic toxicity of soman and other neurotoxicants as well as the efficacy of potential countermeasures was investigated. The link between soman toxicity, glutamate hyperactivity and neuronal death in the central nervous system was investigated in primary dissociated cell cultures from rat hippocampus and cerebral neocortex. Exposure of cortical or hippocampal neurons to glutamate for 30 min produced neuronal death in almost 80% of the cells examined at 24 h. Hippocampal neurons exposed to soman for 15-120 min at 0.1 microM concentration caused almost complete inhibition (> or = 90%) of acetylcholinesterase but failed to show any evidence of effects on cell viability, indicating a lack of direct cytotoxicity by this agent. Acetylcholine (ACh, 0.1 mM), alone or in combination with soman, did not potentiate glutamate toxicity in hippocampal neurons. Memantine, a drug used for the therapy of Parkinson's disease, spasticity and other brain disorders, significantly protected hippocampal and cortical neurons in culture against glutamate and N-methyl-D-aspartate (NMDA) excitotoxicity. In rats a single dose of memantine (18 mg/kg) administered 1 h prior to a s.c. injection of a 0.9 LD50 dose of soman reduced the severity of convulsions and increased survival. Survival, however, was accompanied by neuronal loss in the frontal cortex, piriform cortex and hippocampus. PMID- 7495377 TI - Lipidosis of the dorsal root ganglia in rats treated with an almitrine metabolite. AB - Toxic effects of a detriazinyl metabolite of almitrine (DTMA) were evaluated in rats and on cultured rat macrophages. In rats daily treated with DTMA for 16 weeks, spastic gaits with heel-lifting appeared, and lamellated and/or crystalloid bodies formed in sensory neurons, satellite cells, Schwann cells, and vascular endothelial cells of the dorsal root ganglia. The lysosomal lamellated bodies, which were not induced by almitrine, were produced also in cultured rat macrophages exposed to over 1 x 10(-5) M DTMA. PMID- 7495379 TI - Toxicity of 3-methylindole, 1-nitronaphthalene and paraquat in precision-cut rat lung slices. AB - The toxicity of 3-methylindole, 1-nitronaphthalene and paraquat has been studied in precision-cut rat lung slice cultures. Lung slices were prepared from male Sprague-Dawley rats using an agarose gel instilling technique with a Krumdieck tissue slicer and cultured for 24 h in a dynamic organ culture system. Treatment of rat lung slices with 3-methylindole, 1-nitronaphthalene or paraquat produced concentration dependent decreases in lung slice protein synthesis and potassium content. EC50 values (concentration to produce a 50% inhibition) for protein synthesis were 0.024, 0.27 and 0.57 mM for paraquat, 1-nitronaphthalene and 3 methylindole, respectively. These results demonstrate that precision-cut lung slices are a useful in vitro model system for studying the pulmonary toxicity of xenobiotics. Lung slices offer the potential as a rapid in vitro screen for identifying pulmonary toxicants and to evaluate species differences in response. PMID- 7495378 TI - Acute and subacute inhalation toxicity of diborane in male ICR mice. AB - To clarify the toxicity of diborane, we conducted acute (15 ppm for 1, 2, 4 or 8 h) and subacute (5 ppm for 2 or 4 weeks) inhalation studies on ICR mice. The concentration resulting in a 50% kill after 4 h exposure was 31.5 ppm. Body weight gain was suppressed and the lung weight was increased in diborane-exposed mice in both acute and subacute studies. In the acute study, diffuse pan bronchiolitis-like lesions developed in the lung in various degrees depending on exposure time, which can be pathologically characterized as infiltration of inflammatory cells into the terminal bronchioles and surrounding alveoli, pulmonary congestion and bleeding and/or edema. In the subacute study, we observed lymphoid hyperplasia in the perivascular and peribronchial areas, and infiltration of macrophage and plasma cells into the alveoli. In the mice exposed for 4 weeks, the lesions were more severe than in those exposed for 2 weeks, consisting of hyperplasia and desquamation of Clara cells. In the nasal cavity, we saw mucous exudate and inflammatory cells, suggesting irritation caused by diborane. The histopathological findings, except for the respiratory organs, did not reveal any exposure-related changes. No significant changes were seen in hematological and serum biochemical examinations either. In conclusion, the target organ of diborane inhalation is the respiratory organs, particularly the lung. Further inhalation experiments are essential to investigate the safety exposure levels of diborane. PMID- 7495380 TI - Changes in markers of oxidative status in brain, liver and kidney of young and aged rats following exposure to aromatic white spirit. AB - Levels of glutathione and activity of glutamine synthetase were assayed in organs of rats following inhalation of a heterogeneous solvent mixture containing both aliphatic and aromatic hydrocarbons. This mixture was administered for 3 weeks (6 h daily) at two levels in the inhaled air (400 and 800 ppm) to young adult (5 month-old) and aged (14-month-old) rats. Depression of levels of glutamine synthetase in the P2 fraction of kidney was observed, which was more severe in aged than young adult rats. Glutamine synthetase is a cytosolic enzyme especially susceptible to oxidative damage. A parallel depression of this enzyme was also seen in the corresponding hepatic fractions. However, levels of glutamine synthetase in the hippocampus were elevated by this exposure. Glutathione levels were depressed in P2 fractions of livers of exposed rats, and also in the corresponding renal fraction. Glutathione concentration was unchanged in cerebral fractions. Overall results were interpreted to imply that pro-oxidant events were elevated in kidney and liver following prolonged inhalation of the solvent mixture. The changes found in brain tissue did not reveal evidence of oxidative stress but, however, suggested that glial activation was taking place. PMID- 7495381 TI - Inhibition of intercellular communication by condensates of high and low tar cigarettes. AB - Inhibition of gap junctional intercellular communication (GJIC) is a predictive short term test for tumor promoting activity. A new metabolic cooperation assay has been developed, which takes the cytochrome P-450 metabolism into account. In this assay the inhibitory activity of tobacco smoke condensates (CSC) and CSC fractions from high and low tar cigarettes was tested. CSC of both high and low tar cigarettes and fractions thereof contained tumor promoting activity. The tar yield of the cigarettes did not closely reflect the effects in the GJIC assay and the major constituent nicotine had no effect. The effect was only marginally greater in cells expressing different cytochrome P-450 enzymes, indicating that the active substances are not metabolized by these enzymes. The activities of CSC fractions were considerably lower than the activities in the unfractionated CSC. This may indicate that compounds in the CSC act strongly synergistically. Furthermore, CSC and CSC fractions synergistically inhibit GJIC with the tumor promoter 12-O-tetradecanoylphorbol-13-acetate, indicating different mechanisms of action. PMID- 7495382 TI - Influence of glucose on the toxicity of oxophenylarsine in MDCK cells. AB - Trivalent arsenicals like oxophenylarsine (PhAsO) inhibit cellular pyruvate dehydrogenase, thus leading to a drop of acetylCoA formation and a slow-down of the citric acid cycle. Glucose may protect cells from arsenic toxicity, because increased glycolysis may prevent fatal shortage of ATP. On the other hand, PhAsO has been shown to inhibit glucose uptake in Madin-Darby canine kidney (MDCK) cells. We have investigated the effect of PhAsO on viability, ATP levels and glucose uptake of MDCK cells in the presence of normal (5 mmol/l) and low (0.01 mmol/l) glucose concentrations. At normal as well as at low glucose concentrations, cell viability as assessed by formazan formation was not affected by PhAsO concentrations up to 2 mumol/l within 3 h of observation. At higher PhAsO concentrations viability was diminished earlier and more pronounced in the presence of low glucose concentrations. 10 mumol/l PhAsO induced a drastic drop of ATP within 30 min which was followed by an almost complete loss of viable cells after 180 min in the presence of low glucose concentrations, while at normal glucose levels no influence on ATP contents or on cell viability was detected within 60 min of incubation. On the other hand, glucose uptake, determined as 14C accumulation by cells incubated for 10 min with D-[6-14C] glucose, was inhibited by PhAsO at low as well as at normal glucose concentrations in a dose dependent manner.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7495383 TI - Interaction of phosphamidon with neuropathy target esterase and acetylcholinesterase of hen brain. AB - Phosphamidon (PSM) is an organophosphorus insecticide widely used in agriculture. This study was undertaken to examine the interaction of PSM with acetylcholinesterase (AChE) and neuropathy target esterase (NTE) of hen brain in vitro and in vivo. PSM was a potent inhibitor of AChE, with an I50 of 2.9 microM and second-order rate constant (ka) of 1.2 x 10(4) M-1 min-1 at 37 degrees C. PSM inhibited AChE aged rapidly (t1/2 = 1.9 h). Pyridinium oximes pralidoxime, trimedoxime, obidoxime and HI-6 were effective reactivators of PSM-inhibited AChE, providing up to 75% reactivation. PSM was one of the weakest inhibitors of NTE among organophosphorus compounds, with an I50 of 19 mM and ka of 1.8 M-1 min 1 at 37 degrees C. Inhibited NTE did not reactivate spontaneously and KF-induced reactivation was not obtained even at the earliest tested moments, so it was not clear whether aging of PSM-inhibited NTE occurred very quickly or the KF molecule could not affect the stability of phosphoryl-NTE bond. From the ratio of kas for NTE and AChE (0.00015) it was predicted that delayed neuropathic effects of PSM in vivo would appear only at doses far above the acute LD50. The LD50 value of PSM p.o. for hens was 9 mg/kg. Hens were treated with a single oral dose of PSM, combined with standard antidotal treatment which included atropine, physostigmine, pralidoxime and anticonvulsant midazolam. Doses of 90 and 250 mg/kg caused up to 27% and 45% NTE inhibition 48 h after poisoning, respectively.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7495384 TI - [Role of the clinician in modern cardiology]. PMID- 7495385 TI - [Quality of life]. PMID- 7495387 TI - [High-resolution P-wave electrocardiogram in patients with a history of paroxysmal atrial fibrillation]. AB - PURPOSE: To study by using the signal-averaged P wave, the atrial activation of patients with documented episodes of paroxysmal atrial fibrillation (PAF). METHODS: This study enrolled a total of 20 patients with documented episodes of paroxysmal atrial fibrillation (males 14; mean age 58.4 +/- 10.6 years). The signal-averaged P wave was recorded with a Corazonix Predictor II system. The total P wave duration was determined from the combined filtered x,y,z vector magnitude and used for analysis. The results were compared with a normal group of 10 patients, matched in age. RESULTS: In the control group, the total P wave duration ranged from 120.0 to 135.0 (mean = 128.3 +/- 5.8) ms. In the group of PAF, the total P wave duration ranged from 118.0 to 168.5 (mean = 151.5 +/- 13.7) ms (p < 0.000). Sixteen (80%) of this patients showed a P wave duration > 140.0 ms. With a cut < 140.0 ms for the normal atrial activation, the sensitivity was 76%, specificity was 100%, positive and negative predictive value were 100% and 60% respectively for the method detected patients with PAF. CONCLUSION: Patients with PAF showed a prolonged signal-averaged P wave duration and should be differentiated by this method from the normal population. PMID- 7495386 TI - [The relationship between left ventricular function and the precipitation of symptoms in severe chronic aortic insufficiency]. AB - PURPOSE: To study the relationship between symptoms and left ventricular function in 68 patients with severe chronic aortic regurgitation using echocardiogram and radionuclide left ventriculography at rest. METHODS: Three stages of natural history are assumed: the 1st, at the beginning of the study, when all patients were asymptomatic (clinical stage 0); the 2nd, at the end of 36 month persistence of asymptomatic (clinical stage 1), consisting of 45 patients (group AA); the 3rd, on occasion of manifestation of the symptoms during this period of time (clinical stage 2), consisting of 21 patients (group AS-PRE), with two fatalities not related to valvopathy. RESULTS: The comparison between the groups resulted in similarities in the mean ages, mean time of knowledge of the disease, predominance of males and rheumatic etiology. The profile analysis showed that the significant initial differences between the average of all echocardiographic variables (diastolic diameter, systolic diameter, shortening fraction, final systolic stress, volume-mass ratio, contractility index) and of the ejection fraction of the left ventricle obtained by the radionuclide ventriculography at rest, remained during the study. CONCLUSION: Development of symptoms grouped patients with more advanced excentric hypertrophy, did not coincide with any immediate change in the laboratory markers studied at rest and it was used as a referential for surgical therapy. PMID- 7495388 TI - [Cardiac weight in patients with chronic Chagas disease with Trypanosoma cruzi nests in the central vein of the adrenal glands]. AB - PURPOSE: Comparative analysis of cardiac weight (CW) in patients with chronic Chagas disease, with or without parasitism by Trypanosoma cruzi in the central vein of the adrenal gland (CVAG). METHODS: The CW has been studied in 96 necropsied individuals with chronic Chagas disease. Among them, 48 of them had Trypanosoma cruzi nests on the wall of the CVAG and the other 48 patients did not have nests in the CVAG. Later, this sample was paired for sex, age and anatomo clinical form, resulting in 60 patients. Among these patients, 30 of them had Trypanosoma cruzi nests on the walls of the CVAG and the other 30 did not have. RESULTS: The CW of Chagas patients which had nests of T. cruzi in the CVAG, was 484.8 +/- 146.0g, while in the ones that did not have the nests on the walls of the CVAG, the CW weight was 415.8 +/- 125.3g. This difference was statistically significant. In the first group it had a prevail of congestive heart failure. Subsequently to the pairing for sex, age and anatomo-clinical form, the CW was respectively 464.3 +/- 146.2g and 436.0 +/- 108.9g. This difference was not statistically significant. CONCLUSION: These data suggest that the parasitism in the central vein of the adrenal gland, combined with other possible events intrinsic to the chronic Chagas disease patients' hearts, carries a supporting role in the increasing of the cardiac weight and in the congestive cardiac insufficiency. PMID- 7495389 TI - [Infective endocarditis associated with pregnancy]. AB - PURPOSE: To report the intra-hospitalar and late follow-up of patients with infective endocarditis (IE) acquired in the pregnancy or puerperium. METHODS: Eleven patients, between 1984-1992 according to the beginning of the IE episode (fever and other signals) were studied. Patients were divided in two groups: IE of pregnancy (7 cases), and puerperal IE (4 cases). RESULTS: In the pregnancy IE group, mitral valve was affected in 6 (85%), and aortic valve in 1 (15%). During the course of the treatment, two patients had to be operated on, respectively, in the 24th and 28th week of the pregnancy. The 1st one had a successfully mitral valve replacement by a biological prosthesis but 48 h later she aborted, and the 2nd presented neurological complication (intracerebral hemorrhage) in the immediate post-operative period of a successfully mitral valve replacement by biological prosthesis. She was undergone to a cesarean but the fetus remained alive for 24 h only. If we look at the newborns (fetus), only 4 of them survived. Out of 3 fetal deaths, 2 had close association with mother cardiac surgeries. In the pregnant period acquired IE, 3 (47%) mothers died. In the puerperium group, 2 mitral valves and 2 aortic valves had IE. Two of them had to be operated on due to an important aortic regurgitation and cardiac failure. There were no deaths in this group. CONCLUSION: During pregnancy, IE showed a high morbi-mortality for mother and concept. The mother's neurological complications played a major role in the poor outcome during the pregnancy period. PMID- 7495390 TI - [Ablation therapy of type I atrial flutter with radiofrequency]. AB - PURPOSE--To present initial experience on radiofrequency (RF) ablation of atrial flutter (AFL) guided by anatomic and electrophysiologic parameters. METHODS- Eight patients (six males), mean-age of 42 +/- 17.5 years with chronic type I AFL (mean cycle length of 251 +/- 14.3 msec, range 240 to 280 msec) were undergone to RF catheter ablation applied between inferior vena cava (IVC) and tricuspid annulus (TA). Two had persistent and two the paroxysmal form. Two had surgical corrected congenital heart disease (atrial septal defect in 2 and ventricular septal defect in 1). Four had systolic dysfunction and 2, an atrial tachycardia associated with the AFL. RESULTS--Areas of slow conduction represented by fractionated potentials were recorded between IVC and TA in all patients. RF ablation was successful in 8/8 patients (100%). The mean number of RF applications was 9.2 +/- 6.2 (4-24). The successful ablation site was located in the isthmus between IVC and TA in seven patients and in the lateral wall in the patient with ASD. Successful sites had an early atrial activation preceding the atrial electrogram (range from -65 to -82 ms). In one patient the RF energy was successfully delivered between the atriotomy scar (AS) and IVC. After three months follow-up six remained free of recurrent AFL. One pt had type 1 AFL recurrence and one with ASD had a type II AFL. The Type II AFL was successfully ablated between AS and IVC. CONCLUSION--Fractionated potentials were commonly observed between IVC and T; AFL ablation can be guided by anatomic landmarks or electrophysiologic parameters; electrograms recorded at successful sites were early and never fractionated; the long-term evaluation must be analyzed prospectively. PMID- 7495391 TI - [Profile of the user population of the Emergency Unit of the Heart Institute- HCFMUSP]. AB - PURPOSE: Characterization of the population using the emergency unit of the Instituto do Coracao--HCFMUSP. METHODS: The investigation was carried out in a randomly selected population of 225 people between January and April/93. To determine demographic profile, involvement of the accompanying people with the patient, the nosologic profile and patient's level of knowledge about the Social Health Security (SHS), a questionnaire was applied to the accompanying people. RESULTS: Of the 225 patients, 52.4% were males; age ranged from 1 day to 93 years (45.5% over 60 years old); 61% were living in the state of Sao Paulo; 17% were professionally active; 52% were beneficiaries of Social Security; 92.4% were users of SHS; 47.3% were illiterate; 67% presented with acute cardiovascular diseases and 33% with a noncardiologic diagnosis. CONCLUSION: This is an old population with a low level of instruction, coming from all regions of the country. Because SHS medical services and hospitals does not work well, including cardiology services, this population asked for care in our emergency unit. PMID- 7495392 TI - [Acute hemodynamic effects of ibopamine, captopril, and placebo in patients with severe heart failure]. AB - PURPOSE: To evaluate the acute hemodynamic effects of ibopamine (IBO), captopril (CAP) and placebo (PLA) in patients with severe congestive heart failure at rest. METHODS: Twelve male patients in sinus rhythm with dilated cardiomyopathy and NYHA class IV were studied with Swan-Ganz hemodynamics. Drugs were given in a blinded fashion. Rest, 30 min and every hour for 5 h measurements were made after oral ingestion of 100 mg IBO, 25 mg CAP or PLA. Prior to the study, patients were on diuretics as the only medication for at least 48 h. Comparisons were made with analysis of variance of repeated measurements and Duncan's multiple comparisons procedure. RESULTS: Significant increase in cardiac index and stroke volume index and reduction in systemic vascular resistance were observed with IBO and CAP for 2 h after ingestion. IBO however increased right and left filling pressures in the first hour after its administration. Ventricular tachycardia occurred in 2 patients 1 h after IBO administration. CONCLUSION: Both IBO and CAP improved hemodynamic parameters in the first two hours after oral ingestion in patients with dilated cardiomyopathy in class IV. PMID- 7495393 TI - [Anomalous origin of the circumflex artery and coronary spasm]. AB - A 43 year-old female patient with angina pectoris and vasospasm demonstrated in the anomalous left circumflex (Cx) and right coronary (RCD) arteries by coronary angiograph. Origin of the left Cx from the RCD is the most common coronary anomaly and generally, is considered to be benign. Nevertheless, myocardial ischemia in patient with this anomaly has been described. To our knowledge, this is the first reported case of coronary vasospasm occurring simultaneously in the anomalous left Cx and RCD arteries. The diagnostic troubles and the potential danger of this association were emphasized. PMID- 7495394 TI - [Effects of thrombolytic therapy on mortality of inpatients at adult intensive care units at the Municipal Hospital]. AB - The lack of facilities sometime blocks the use of thrombolytic therapy in public hospitals. In this report, initial experience with the use of thrombolytics in a Municipal Hospital located in Sao Paulo suburb area, which has no hemodynamic laboratory available, is reported. Prospectively, 45 patients admitted to the adult medicine care unit with acute myocardial infarction from Dec/93 to May/94 were evaluated. Twenty four patients were submitted to thrombolytic therapy by streptokinase (SK), 1.500.00 units IV in the peripheral vein. They showed the following signs of reperfusion: relief of pain, rapid resolution of ST-segment elevation, reperfusion associated arrhythmias and early enzymatic peak (CK-MB). Seven deaths were recorded, six of them did not receive SK. The use of thrombolytic, even in hospitals without haemodynamic laboratory, has contributed to the decrease of mortality. Thus, this therapy should be used as early as possible in acute myocardial infarction. PMID- 7495395 TI - [Congestive heart failure associated with ischemic cardiopathy]. PMID- 7495399 TI - [Noninvasive evaluation of ischemia and viability after acute myocardial infarction]. PMID- 7495397 TI - [Symptoms and ventricular dysfunction in chronic aortic insufficiency]. PMID- 7495398 TI - [Advances in echocardiography. Introduction]. PMID- 7495396 TI - [Menopause, hormonal replacement therapy, and coronary disease]. PMID- 7495400 TI - [Use of contrast echocardiography in coronary insufficiency]. PMID- 7495401 TI - [Transesophageal echocardiography. Indications and clinical impact]. PMID- 7495402 TI - [Evaluation of systolic and diastolic left ventricular function by echocardiography]. PMID- 7495404 TI - [Hemodynamic evaluation of congenital heart defects by Doppler echocardiography]. PMID- 7495403 TI - [Stress echocardiography. Rise of a new method]. PMID- 7495405 TI - Health care changes bring increased liability risk for nurses. PMID- 7495406 TI - Who's taking care of mama? PMID- 7495407 TI - [Skeletal findings in an adult dog recovering from osteodystrophia fibrosa]. AB - In a 9-year-old male German Shepherd dog with a body weight of 37 kg which was euthanised because of a spinal meningioma, examination of the carcass revealed severe signs of degradation concerning the whole skeleton which were investigated by radiological, microradiological and pathologic-histological means. The skeletal alterations in form of systemic defects in corticalis and spongiosa in fact correspond to appearance and localisation of bone defects typical of fibrous osteodystrophia, but now are filled by conspicuous cell-poor matured collagenous connective tissue. In this connection it must be emphasised, that there are no osteoclasts detectable in the lacunae of degradation so that the disease process apparently has stopped. PMID- 7495410 TI - [Acute myeloid leukemia (promyelocytic leukemia) with detection of virus particles in a boar. Histologic, histochemical and ultrastructural findings]. AB - Acute myeloid leukemia is a rare finding in swine. A case of acute promyelocytic leukemia is described by light microscopy, enzyme- and immunohistochemistry as well as electron microscopy. Tumour cells can be demonstrated in bone marrow, lymph nodes, liver, spleen, kidneys, small intestine and myocardium. The tumorous neutrophil precursor cells are positive for naphthol-AS-D-chloroacetate esterase and lysozyme. They show ultrastructurally numerous granules as typical for promyelocytes. Virus particles of the oncorna-c-type can be demonstrated within the tumour cells. PMID- 7495412 TI - [The content of pepsinogen in the blood plasma of growing fallow deer (Dama dama L.) raised helminth free and under natural conditions in captivity]. AB - Pepsinogen values were determined in helminth free (n = 12) and naturally raised farmed fallow deer (n = 10-12) from birth (June 1992) to 11 (May 1993) and 15 months (September 1993) of age respectively in 4 weekly intervals. At birth the plasma pepsinogen value was 0.708 +/- 0.197 U Tyrosin/l and decreased thereafter until 14 weeks of age to 0.139 +/- 0.061 U Tyrosin/l in both groups. From the 18th week of age the naturally raised fallow deer, infected with gastrointestinal nematodes, had significantly higher pepsinogen values than the helminth free kept animals. Maximum pepsinogen levels in the naturally raised fallow deer were determined in November 1992 (0.813 +/- 0.336 U Tyrosin/l) and in June 1993 (1.050 +/- 0.163 U Tyrosin/l). In helminth free animals no further decrease of plasma pepsinogen values was seen from 14th week of age onwards: it was 0.202 +/- 0.110 U Tyrosin/l. There was no influence of sex on the plasma pepsinogen values in both groups. PMID- 7495411 TI - [The significance of vitamin A for the immune system (review)]. AB - The significance of vitamin A and of its metabolites (retinoic acid a. o.) for the functional capacity of the cells of the immune system (macrophages, T- and B lymphocytes, killer-cells a. o.) is discussed with special consideration to that of the gastro-intestinal-tract and of the mammary gland. Recommendations for the application of vitamin A in combination with the vitamins D, E and C are given. PMID- 7495413 TI - [Artificial lactation induction in 12- to 19-month-old heifers with regard to the behavior of the concentration of sex hormones and lactogenic proteohormones in the blood plasma]. AB - In 24 prepubertal heifers the lactation was artificially induced by two different methods of hormone application. The changes in milk yield and the concentrations of estradiol-17 beta (E), progesterone (P), prolactin, somatotropin and insulin like growth factor I in blood plasma during the first 55 days of lactation were analyzed. Depending on the age, the animals were divided in three experimental groups (EG) of eight animals each: 12 months old (EG I), 15 months old (EG II) and 19 months (EG III). Four animals of each EG were treated similarly: group E and P, which was injected by estradiol benzoate and P for 7 days, or group E, which received only estradiol benzoate for 10 days. The oldest heifers (EG III) showed an induction rate of 100% with E and P treatment and of 75% with E alone. On the other hand, in heifers of EG I and EG II the induction rate was only 50% irrespectively of the treatment. The animals of EG III had the highest milk yield. The daily milk yield on the 50th day of lactation was 7.0 kg and 4.0 kg in group E and P or group E, respectively. Comparable levels in 15 months old heifers (EG II) were 4.8 kg and 1.4 kg, and in 12 months old heifers (EG I) 2.5 kg and 2.5 kg. The changes of the concentrations of the hormones in blood plasma are demonstrated in diagrams, their effect on the milk production is discussed. PMID- 7495408 TI - [Fertility in mares after disturbed or undisturbed puerperium; evidence from clinical, microbiologic and hormone analysis]. AB - Because of the mare's ability to conceive very soon post partum a thoroughly performed control of the puerperal period is of particular importance. In addition to cytological and histological examinations of the uterus, rectal palpation and vaginoscopic inspection as well as microbiological examinations of uterine swabs and the evaluation of the mare's hormonal status during the puerperal period provide useful information which can be used either prognostically or to initiate a specific therapy. 55 mares were examined on days 3, 6 and 9 post partum. Rectal palpation, vaginoscopic inspection, microbiological examination of uterine swabs and determination of progesterone (P4)- and estradiol-(E2)-levels in the peripheral blood were performed on all the three days. On day 18 after the last breeding the mares were checked for pregnancy by ultrasonic echography. We observed many cases of heavy bacterial contamination of the uterine contents during the puerperal period up to the foal heat. Especially coliforms and beta-hemolytic streptococci were often found. However, these contaminations could not be proven to interfere with fertility rates during the first post partum estrus periods. Dystocia and retained placenta had a strong influence on the hormonal situation of mares during the puerperal period. In addition to this we found disturbed involution of the genital system in these mares. Much higher pregnancy rates at foal heat were detected after a rapid onset of ovarian activity post partum especially during May, June and July, compared to mares with delayed onset of ovarian activity.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7495409 TI - [The effect of calcium 2+ ions on the decomposition of canine and human fibrin(ogen) by (human) plasmin]. AB - With a row of degradation kinetics with different Ca2+ concentrations and their analysis by the means of nonreducing SDS-PAGE we investigated the dependence of fibrin(ogen) degradation product patterns on the Ca2+ concentration. At man and dog the addition of calcium stabilized from 0.06 mM Ca2+ (2.0 molecules Ca2+/molecule Fibrinogen) certain X- Y- and D-fragments which were not or not in this extent generated by degradation without calcium. The X- and Y-fragments were degraded further by plasmin cleavage at other positions, the greatest D subfragment (D1) remained stable. As like as known for human fibrinogen the Ca2+ bound on a D gamma-chain position probably prevented the cleavage of a C-terminal part of the gamma-chain. By the addition of growing Ca(2+)-concentrations (from 0.1 mM to 10 mM Ca2+) an additional, increasing D-dimer spot at a molecular weight of 220 +/- 7 kDa was formed owing to progressive activation of the concomitant calcium-dependent transglutaminase (factor XIII). After complete proteolysis of fibrinogen (after 15 min) we observed the formation of D-dimers from canine D1-fragments. At the fibrin degradation the D-dimer was dominating already from 0.1 mM Ca2+ in the degradation assay. Especially the D-fragments, but also the smaller FDP E an -F were generated here in a reduced extent. Several new bands (= 57 kDa) were formed instead, that were probably connected FDP D, -E and -F, crosslinked through the action of transglutaminase.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7495415 TI - [The acid-base balance in the blood of calves of different breeds in a mother cow herd raised on deep straw and strawless conditions]. AB - The influence of two different animal loose housing systems (deep litter resp. gum lying area/slatted floor) with additional calf pen on blood gases and buffer systems in the blood of 19 calves of the breed Deutsche Schwarzbunte and 20 calves of the breed Deutsche Rotbunte of both sexes in a suckler cow herd was examined in a period from the first to the sixth month of life. The pH of the blood remains within the well-known small limit of variability, and no exogenic influences could be seen. Age dependent course up to the third or fourth month of life found out in the factors of oxygen content and oxygen transport, which reflect the development to adults. The pCO2 is higher in calves kept on slatted floor, and the difference is significant in the second and sixth month of life. No significant differences found out in buffer base between the loose housing systems but a slow decrease is perceptible with the development to ruminant conditions. In the behaviour of calves no significant differences could be seen but calves on slatted floor had some more distance to their mothers. No sex and breed specific differences found out in behaviour or physiological data. In conclusion, an extensive suckler cow management in different equipped loose housing systems is practicable provided, the calves have a separate calf pen. PMID- 7495416 TI - [Behavior disorders in relation to food intake in African ostriches]. AB - The behaviour of ostriches (Struthio camelus) was observed. The most striking behaviour disorders was feather pecking. However, the animals showed four other behaviour patterns, which also must be qualified as behaviour disorders: Pecking sand, pecking wire, pecking their own feathers and pecking into air, occurred in a random succession. This is a result of the nutrition, which differs from the natural feeding situation, as the animals are fed exclusively on pellets so that the food consumption is completed within a very short time. This results in an accumulation of action specific energy within the functional circle of nutrition. Therefore in the husbandry of ostriches in Central Europe not only the necessary energy and sufficient composition of nutrition should be considered. It is necessary as well, that the food be of a consistency which guarantees food intake of appropriate satisfactory duration. PMID- 7495414 TI - [The composition of milk after artificial lactation induction in 12- to 19-month old heifers]. AB - In 24 prepubertal heifers the composition of milk after an artificial induction of lactation was investigated. Depending on the age, the animals were divided in 3 experimental groups (EG) of 8 animals each: 12 months old (EG I), 15 months old (EG II), and 19 months old (EG III). Four animals of each EG were treated similarly: either with estradiol benzoate and progesterone for 7 days (group E and P) or only with estradiol benzoate for 10 days (group E). During the first 18 days of lactation the protein and lactose content of the milk showed similar changes compared to the period of milk maturation during natural lactation of same duration. However, a typical colostral period with high concentrations of immunoglobulins in the milk was not observed. After the artificial induction of lactation, in the period until to the 53rd day of lactation no age dependent differences in the composition of milk were found, except whey proteins. Formation of milk substances, which are synthesized in the mammary gland, is much more induced by treatment with E and P than by treatment of E alone, with the exception of caseins. PMID- 7495418 TI - Space-occupying lesions of the sensori-motor region. AB - Successful surgery of the sensori-motor region requires precise pre- and intraoperative localization of the sensori-motor region and pyramidal tract. Important aids are the landmarks of cranio-cerebral topography, coronal suture and bregma and the sulcal anatomy of the sensori-motor region, which can be identified in CT or MR images. Due to considerable displacement and distortion of the anatomical structures, elicited by mass lesions, these aids often fail to render reliable support. In this situation, identification of the motor area can be achieved by electrical stimulation of the precentral gyrus in association with the recording of somatosensory evoked potentials of the pre- and postcentral gyrus. The localisation of the "motor mosaics" in relation to the lesion, enable determination of the direction of displacement of the motor strip and the fan of the pyramidal tract. Based on this information the most appropriate route of access to the lesion is selected, either transcortical or transsulcal. Lesion specific operative techniques as well as location-specific approaches are discussed. With consequent application of these principles the risk of a new persistent motor deficit was as low as 4%. Thus, the indication for surgery in this area can now be set with greater confidence and far more generously than in the past. PMID- 7495417 TI - [Differentiation of Salmonella gallinarum rough and smooth strains using gas chromatography analysis of their cell-bound fatty acids]. AB - The whole cell fatty acids of 7 field isolates of smooth (S)-type S. Gallinarum strains and 3 isolates of rough (R)-type S. Gallinarum strain 9R were determined by the liquid-gas-chromatography using the Microbial Identification System (MIS, Microbial ID, Inc. [MIDI, Newark, Delaware USA]). The profiles of the bacterial cellular fatty acids of the S- and R-type isolates differ in the number as well as in the quantity of the fatty acids. On the basis of these differences the S type isolates could be distinguished from the R-type cultures. PMID- 7495419 TI - The surgery of cavernomas both supra-tentorial and infra-tentorial. PMID- 7495421 TI - The classification and molecular biology of pituitary adenomas. PMID- 7495420 TI - Surgery for gliomas and other mass lesions of the brainstem. PMID- 7495422 TI - Hearing preservation in acoustic tumour surgery. AB - 900 acoustic neurinomas were removed by the suboccipital approach at Nordstadt Neurosurgical Department from 1978 to 1992 by the same surgeon (M. S.). While 247 patients were deaf on the involved side before surgery, there were 653 patients ears with some preoperative hearing. Preservation of the cochlear nerve was always attempted, and the overall-rate of hearing preservation was 38% (249 of 653), regardless of pre- and postoperative quality of hearing or of tumour sizes. In small tumour sizes below 3 cm of diameter preservation rate was 51%, in large tumours above 3 cm of diameter it was 22%. A classification system of hearing quality was made up considering pure tone audiometric hearing losses (PTA HL) at 1 to 3 kHz, and individual maximum speech discrimination scores. The usefulness of the preserved hearing is further evaluated considering the quality of hearing in the contralateral ear, and by application of other classification schemes. Presentation of the surgical strategies and their refinements by personal experience provide the base for discussion questioning whether and how further progress may still be anticipated. PMID- 7495423 TI - Biomechanics and classification of traumatic lesions of the spine. PMID- 7495424 TI - In vivo measurement of spinal column viscoelasticity--an animal model. AB - The goal of this study was to measure the in vivo viscoelastic response of spinal motion segments loaded in compression using a porcine model. Nine pigs were used in the study. The animals were anaesthetized and, using surgical techniques, four intrapedicular screws were inserted into the vertebrae of the L2-L3 motion segment. A miniaturized servohydraulic exciter capable of compressing the motion segment was mounted on to the screws. In six animals, a loading scheme consisting of 50 N and 100 N of compression, each applied for 10 min, was used. Each loading period was followed by 10 min restitution with zero load. The loading scheme was repeated four times. Three animals were examined for stiffening effects by consecutively repeating eight times 50 N loading for 5 min followed by 5 min restitution with zero load. This loading scheme was repeated using a 100 N load level. The creep-recovery behavior of the motion segment was recorded continuously. Using non-linear regression techniques, the experimental data were used for evaluating the parameters of a three-parameter standard linear solid model. Correlation coefficients of the order of 0.85 or higher were obtained for the three independent parameters of the model. A survey of the data shows that the viscous deformation rate was a function of the load level. Also, repeated loading at 100 N seemed to induce long-lasting changes in the viscoelastic properties of the porcine lumbar motion segment. PMID- 7495425 TI - Clamping the small intestine during surgery: predicted and measured sealing forces. AB - During bowel surgery it is often necessary to occlude the bowel using clamps. Occlusion occurs in two stages: approximation of bowel internal surfaces followed by sealing which is necessary to prevent seepage. The occlusive force at both stages depends on the luminal pressure, the latter stage requiring greater force than the former. The difference in luminal pressure across the sealing line results in a slipping force which is resisted by friction between the bowel and the clamp jaw surfaces. A theoretical model was developed to describe these forces and the predicted values obtained from this model were compared with direct measurements carried out on porcine bowel samples in a test rig. It was found that the measured approximation force was between 25 and 50 per cent of the maximum theoretical values and that sealing without seepage requires a clamp force of about ten times the approximation force. Using these results and known intraluminal pressures in the human gastrointestinal tract, a bowel clamp must apply around 7 N to prevent seepage and the coefficient of friction between clamp and bowel should be between 0.6 and 0.9. PMID- 7495428 TI - A measurement system for the recognition of arm gestures using accelerometers. AB - This paper describes a strategy to measure arm movements using accelerometers for the computer recognition of arm gestures. Gesture recognition is being investigated as an alternative method of computer input for people with severe speech and motor impairment; the emphasis is on the needs of people with athetoid cerebral palsy who have difficulties with existing computer input devices. An initial model-based approach to estimate the kinematic motion of the arm from acceleration measurements is given, followed by the chosen measurement scheme. The current system considers the forearm as a rigid body and uses two data streams derived from four linear accelerometers. By treating these signals as outputs from postulated mechanical models, the data are reduced to two series of step inputs that are appropriate for pattern classification and recognition. Although this does not in any way model the arm, the concepts used are based on the type of driving signals expected in the control of arm gestures. Initial experimental results show that the information content of gestures is preserved by this data parameterization. PMID- 7495426 TI - Detachable clamps for minimal access surgery. AB - A detachable clamp and applicator have been developed for use in minimal access surgical operations involving hollow visceral transection and anastomosis. The clamp has parallel jaws which ensure uniform distribution of the occlusive force. Following application on the bowel, the clamp is released from the applicator, thus freeing the access port. On completion of the anastomosis, the clamp is docked to the applicator, its jaws opened for release from the bowel and then closed prior to removal. The jaws of the clamp are kept closed by a pseudoelastic nickel-titanium (NiTi) alloy spring which imparts advantageous force characteristics when compared to stainless steel. The excellent holding and atraumatic characteristics of the detachable clamp have been confirmed by use in laparoscopic and thoracoscopic surgery on the gastrointestinal tract. PMID- 7495427 TI - Determination of the curvatures and bending strains in open trileaflet heart valves. AB - The leaflets of trileaflet artificial heart valves manufactured from polyurethane, gluteraldehyde-treated porcine aortic valves and pericardial tissue are subject to cyclic stresses and strains which can reduce the lifetime of the implanted valves through leaflet calcification and fatigue failure. A detailed knowledge of the stress state within a valve leaflet throughout a cardiac cycle is desirable in order to improve the geometry of the valve leaflets and ultimately improve the valve performance. An experimental method to evaluate the radius of curvature at the free edge of the open valve leaflet is presented. The technique has been applied to polyurethane trileaflet heart valves manufactured within the authors' laboratory and to commercially available bioprosthetic valves in the fully open position under steady and pulsatile flow conditions. Simple bending theory has been applied to the polyurethane valves to calculate bending stresses and strains at the free leaflet edge based on the measured curvature. The results showed that in the fully open position the highest curvatures occurred at the commissural regions for all the valves analysed. Additional areas of high curvature were present along the free leaflet edge. Average curvatures as high as 0.85 mm-1 were observed at the leaflet commissures for the polyurethane valves with a resultant bending stress of 0.72 MPa. The porcine bioprosthetic valves showed average curvatures as high as 2.5 mm-1 which also occurred at the leaflet commissures. The results of the study have been compared to values of stress obtained from numerical analysis of closed polyurethane valve leaflets reported in the literature.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7495429 TI - Cushion form bearings for total knee joint replacement. Part 1: Design,friction and lubrication. AB - Cushion knee prostheses have been designed and constructed that produce approximately equal initial contact areas and theoretical film thicknesses compared with a conventional UHMWPE (ultra-high molecular weight polyethylene) joint. These compliant bearings had a flat tibial component which imposed fewer biomechanical constraints and allowed a greater range of movement. Friction experiments have been carried out on a pendulum simulator apparatus. The results showed that the cushion knee joints operated just within the mixed lubrication regime, but that they benefited from a substantial measure of fluid film lubrication. Microelastohydrodynamic lubrication was effective in preserving low friction and thin but effective lubricating films. PMID- 7495430 TI - Cushion form bearings for total knee joint replacement. Part 2: Wear and durability. AB - Cushion knee prostheses have been designed and constructed to produce larger initial contact areas and thicker theoretical film thicknesses than a conventional UHMWPE (ultra-high molecular weight polyethylene) joint. The compliant bearing had a flat tibial component which imposed fewer biomechanical constraints and allowed greater range of movement. Wear tests were performed in a knee joint simulator and creep tests were carried out in a servo-hydraulic apparatus. Various failure modes of cushion joints that require further study were identified. However, the results showed that adequate durability was achieved from a 20 MPa polyurethane material in joint simulating tests carried out over 0.5, 1.0 and 5.0 million cycles. Most importantly, during these tests, no detectable wear debris was generated. It is believed that this is the first time that the full potential of cushion bearings has been demonstrated in a joint simulator over periods corresponding to about five years of service in vivo. PMID- 7495431 TI - An estimation of fatigue life for a carbon fibre/poly ether ether ketone hip joint prosthesis. AB - A fracture mechanics approach was applied to estimate the life of a prosthesis injection moulded from short carbon fibre reinforced poly ether ether ketone. Flexural modulus and strength, fracture toughness, fatigue endurance limit, fatigue crack growth rate and threshold stress intensity factor were determined. The dimensions of the test pieces were selected to yield fibre orientation and fibre length distributions similar to those obtained in the prosthesis. Stress levels generated in the prosthesis under different activities were estimated by conducting three-dimensional finite element analysis. It was shown by a fracture mechanics approach that a fatigue failure due to the propagation of an embedded elliptical slit, under these stresses, would be unlikely for a crack length smaller than 1.85 mm. However, the cement would fail under the same conditions, irrespective of the type of the prosthesis employed. PMID- 7495432 TI - [Changes in the auditory threshold for air and bone conduction in relation to middle ear pressure in probands with normal hearing]. AB - Under pressure in the tympanic cavity causes increased impedance of the middle ear. Gelle was the first to describe increased bone conduction levels following alteration of ear canal pressure in healthy ears. Up to now, no investigation which quantitatively describes the elevation of the hearing threshold induced by various levels of under pressure in the middle ear has been published. In a pressure chamber, we induced relative under pressure in the middle ears of 15 adults with normal hearing. We measured hearing thresholds and calculated medium values at four separate levels of under pressure. At an under pressure of 3.3 kPa, air conduction was reduced by a few dB at 500 and 1000 Hz. Alterations of bone conduction were first seen at 6.6 kPa accompanied by increased deterioration of air conduction. Both effects became more obvious at 10 kPa; and at a maximum under pressure of 13.3 kPa, a deterioration of air conduction by more than 25 dB was seen at 250, 500, and 1000 Hz. Bone conduction deteriorated by more than 10 dB at 500 and 1000 Hz. There was no uniformity in the development of bone conduction threshold in the condition of under pressure: Several ears expressed only slight changes, but in some ears we saw an increase of bone conduction at the same rate as air conduction. Minor alterations were observed in frequencies above 1000 Hz. These results may be only partially explained by middle ear effects like the reduction of the ostio-tympanic component of bone conduction caused by increased stiffness of the ossicles. We believe that disturbances of inner ear mechanics play a role in the deterioration of bone conduction levels, too.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7495433 TI - [Diagnosis of auditory threshold with frequency-specific recording of auditory evoked potentials: personal results and methodological aspects]. AB - Results and Overview of Methods: Auditory brain stem responses are widely used in objective assessment of hearing function in both children and adults. Because of the lack of frequency specificity, several attempts were made to optimize responses using tone stimuli. Tone stimuli are characterized by intensity, frequency, and duration. Hence, there are important influences on the waveforms with respect to rise, plateau, and fall time. In addition, noise masking techniques are used to suppress both spectral sidebands of the stimulus and cochlear response not corresponding to the test frequency. In this paper, results on tone burst stimulation in notched noise masking are presented. Using this technique, reproducible waveforms (Jewett I to V resembling peaks) including a middle latency component could be obtained. Various stimulus characteristics (clicks, tone pips, tone bursts, Gaussian shaped stimuli), masking techniques (high-pass noise, notched noise), spectral characteristics, and time saving strategies are critically discussed. PMID- 7495434 TI - [Reliability of brain stem audiometry in specific learning disorders (disorders of sensory integration)]. AB - Diagnostic audiological tests performed in ten children from 1985 to 1991 showed remarkable differences between the results of behavioral audiometry (free field or pure tone audiometry) and those of auditory evoked response audiometry. Auditory brainstem response (ABR) was absent in seven children with sensory neural hearing loss. Neuropsychological evaluation was performed on four children, neuropsychological observation was performed on two children, pediatric examination was performed on three children and one child showed signs of neuropediatric disorder. The results of neuropsychological evaluation showed moderate to severe learning disabilities (sensory integration disorders, learning disorders) in all four children tested. Available neuropsychological observation also indicated the presence of sensory integration disorders in two children. Two children had multiple handicaps with cerebral abnormalities, two further children were diagnosed as suffering from "minimal cerebral dysfunction," one of which was mentally retarded. The fact that ABRs were absent in seven children indicates that a response desynchronisation in the auditory pathway may exist in these disorders. Thus, ABR might not be a reliable method for audiological testing in children with learning disabilities or other cerebral dysfunctions, but serves as an adjunct to conventional testing methods. PMID- 7495435 TI - [Hearing screening within the scope of pediatric U5 preventive examination: can diagnosis be improved by use of a parent questionnaire?]. AB - OBJECTIVE: Assessment of a) the efficacy of hearing loss screening in baby check ups at the age of 6 to 8 months b) the efficiency and efficacy using parent questionnaires about hearing loss risk factors and parents' concern about the child's hearing as part of baby check-up. METHODS: The analyses are based on 7282 baby check-ups carried out in 47 pediatricians' offices in the Dusseldorf area from July 1991 to March 1993: 3385 of these check-ups were performed in the traditional manner (period A; July 1, 1991, to January 1, 1992; 3897 check-ups involved parent questionnaires (period B June 1, 1992, to March 31, 1993). All children who failed the tests and a random sample of those who had passed the test when tested in the traditional manner, were offered extensive hearing assessment. RESULTS: In period A, 4.3% of the children failed the test as compared to 21.7% with the first version of the questionnaire and 14.3% with a revised questionnaire in period B. Compliance to the offer of extensive hearing assessment was unsatisfactory (53% in period A and 63% in period B). The first extensive hearing examination failed to distinguish healthy children from children with probable sensorineural hearing loss in about 30% of the patients, mainly because of glue ear. One child with sensorineural hearing loss was identified by the screening in period A as compared to six children in period B. CONCLUSIONS: 1) The systematic use of parent questionnaires in baby check-ups at ages 6 to 8 months is likely to increase the efficacy of the screening. 2) The efficiency of the questionnaires currently in use, however, needs to be improved and can be improved with the data from the study. 3) The compliance with extensive hearing testing in children who have failed the screening test must be improved. 4) Due to the high prevalence of conductive hearing loss in this age group, treatment of potential glue ear problems is mandatory before referral for extensive testing for sensorineural hearing loss. PMID- 7495436 TI - [Facial nerve abnormalities in malformed temporal bone]. AB - Between 1987 and 1993, 4500 tympanoplasties and stapedectomies were performed at the ENT Department of the University of Wurzburg. In 134 cases, surgery was performed for malformation of the middle ear. The surgery reports of these patients were analyzed for anomalies of the facial nerve. Forty cases (29.9%) showed an anomaly of the facial nerve, more often congenital aural atresia or major malformation (30.4%) than minor malformation (29.1%). The most frequent finding was a dislocation of the nerve over the oval niche and resulting narrowing of the oval window (22 cases). Eleven cases showed the facial nerve without bony covers, in two cases there was an exposed nerve over the oval niche. An acute-angled course between horizontal and vertical segment in the mastoid, doubling of the nerve, lateralization of the mastoidal segment, and anterior shift of the Fallopian canal in the mastoid were observed once. To reduce the risk of facial nerve injury, high resolution CT scan studies should be performed before surgery in the presence of congenital aural atresia. PMID- 7495437 TI - [Detection of serum IgA antibodies to varicella zoster virus (VZV)--differential etiology of peripheral facial paralysis. A case report]. AB - During the early manifestation of peripheral facial paralysis, VZV specific IgA antibodies were detected in the serum in a single case. In similar cases, this serological parameter should be tested for causal diagnosis in connection with early Aciclovir therapy. PMID- 7495438 TI - [Effects of surgical therapy of obstructive snoring on swallowing and voice quality]. AB - The efficacy of uvulopharyngopalatoplasty (UPPP) in treating obstructive snoring has been reported from many centers. The reduction of uvula and soft palate may result in palatal incompetence. We examined 23 patients undergoing UPPP pre- and postoperatively and found a slight preoperative hypernasality in three patients. None of our patients had problems with swallowing. During the first week postoperatively, most of the patient had hypernasality and noted a nasal escape of liquids. Three to four months after surgery, three patients still had problems swallowing. All 21 patients seen during the last follow-up examination had a nasal escape of air detected by auscultation in at least one of the tests. Two patients had an audible hypernasality. Two patients could not pronounce the letters any more. More than one year after surgery, palatal incompetence was no longer detectable. PMID- 7495439 TI - [Cultivation of human cartilage tissue in a 3-dimensional perfusion culture chamber: characterization of collagen synthesis]. AB - In reconstructive head and neck surgery, there is a great need for cartilage transplants. Sufficient autologous graft is often not available. Heterologous cartilage is used frequently, although there is danger of transmitting viral infections and resorption rates are high. We have developed a three-dimensional model for the formation of cartilage in vitro. The aim of this study was to characterize the collagen synthesis under these culture conditions. Human chondrocytes were isolated by digesting septal cartilage matrix in the presence of type II collagenase, hyaluronidase, and Dnase II in Ham's F12 medium. The resulting cells were kept in monolayer culture for one week and then suspended in 2% ultra-low-melting agarose (1:1). The cell-agarose conglomerate was encapsulated with a 3% ultra-low-melting agarose solution and placed in a perfusion culture chamber. A permanent flow of fresh medium (Ham's F-12 supplemented with 50 micrograms/ml ascorbic acid and 2% fetal calf serum) was provided by a peristaltic pump which delivered 1 ml/h with on/off intervals of 30 min. Samples were recovered after two weeks. Using electron microscopy abundant collagen fibril formation was shown. The collagen fibrils were identified histologically as cartilage specific type II collagen. No mRNA expression of collagen type X was observed using in situ hybridization. The cells appeared in a round cell shape with round nucleus and only slight variations in form and size. The present results indicate that the chondrocytes maintain their differentiated phenotype and continue to synthesize typical matrix products in this three dimensional perfusion culture chamber.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7495440 TI - [In vitro studies of the effect of vasoconstrictor nose drops on ciliary epithelium of human nasal mucosa]. AB - We investigated the influence of four nasal decongestants and one preservative on ciliated cells, using cell cultures of human nasal mucosa. These cells were exposed to nasal decongestants in vitro. A fast decrease in ciliary beat frequency was seen during naphazoline and oxymethazoline application which was fully reversible after naphazoline. There was only a minimal decrease in ciliary beat frequency during application of phenylephrine and xylomethazoline decongestants. The preservative benzalkoniumchloride caused an irreversible decrease in ciliary beat frequency which was concentration dependent. We therefore recommend nasal decongestants which contain xylomethazoline or phenylephrine. If the preservative benzalkoniumchloride can not be substituted by newer less harmful substances, it should be added in minimal concentrations. PMID- 7495442 TI - [Churg-Strauss syndrome. A differential diagnosis in chronic polypoid sinusitis]. AB - BACKGROUND: The Churg-Strauss syndrome (CSS) is a rare multiple organ disease, that belongs to the group of systemic granulomatous vasculitis. The initial symptoms are often bronchial asthma and allergic rhinitis; later in the course of the disease, the patients exhibit lung, heart and kidney manifestations. PATIENTS: We report on a thirty-one year old female patient who initially presented with asthma and polypous sinusitis. Three years later, after undergoing endonasal sinus surgery three times, she was diagnosed with CSS. Within a review of the literature, we discuss symptomatology, differential diagnosis, and therapy of the CSS. RESULTS: Worldwide about 200 cases are published. The patient presented in this case report is now in a good health condition after a four-year therapy with steroids. The patient is free of sinusitis symptoms without the need of renewed surgical intervention. CONCLUSIONS: CSS should be considered in the differential diagnosis if bronchial asthma and resistant polypous sinusitis occur coincidentally. Often it is very difficult to diagnose CSS. Frequently it is only diagnosed after several years. Therapy of choice is the treatment with steroids, sometimes supplemented by cytotoxical drugs. PMID- 7495441 TI - [Morphology and histochemistry of lymphatic vessels of the upper aerodigestive tract: a clinically oriented study]. AB - BACKGROUND: Exact knowledge of lymphatic morphology and histochemistry is required to understand the clinical importance of the lymphatic system of the upper aerodigestive tract. Publications in this field are rare. METHODS: Light and electronmicroscopic examinations were carried out on lymphatics of the upper aerodigestive tract. Enzyme, immune and histochemical investigations were also performed on 764 tissue specimens from this region. RESULTS: The discontinuous basement membrane surrounding the initial lymphatic contains laminin, type IV collagen, and fibronectin and is usually continued by a fibrillar elastic apparatus that is less developed than dermal lymphatics. 5'-nucleotidase, adenylatcyclase, and guanylatcyclase activity in lymphatics is not as high as it is in blood capillaries. The endothelium of collecting lymphatics contains high concentrations of factor VIII associated antigen, while it is found in the endothelium of initial lymphatics only in small amounts or not at all. Histochemical studies of the endothelium of lymphatics show a positive lectine binding reaction for UEA I, PNA, DBA, GS I, MPA, and RCA I. CONCLUSIONS: The histochemical results obtained allow easy differentiation of lymphatics and blood capillaries. The vessel type differentiation is relevant for the examination of organ-related lymphatic systems, to determine whether tumor cell nests are located in lymphatics, blood capillaries, or artificial tissue gaps. PMID- 7495443 TI - [Suppurative parotitis as a sequela of infection with Salmonella enteritidis]. AB - Extraintestinal manifestations of Salmonella infection are rare events and occur primarily in immunocompromised hosts. We report on a 69-year old male patient hospitalized for absceding suppurative parotitis. From the swab taken on incision of the abscess, Salmonella enteritidis was cultivated. There was no preceding gastrointestinal infection in our patient. Risk factors in our patient included a poorly controlled non-insulin-dependent diabetes mellitus, chronic alcohol abuse, and advanced age. Quinolones are the therapy of choice in extraintestinal salmonellosis. On ciprofloxacin perorally, the patient showed rapid recovery. PMID- 7495444 TI - Are VA hospitals necessary? PMID- 7495445 TI - Elder law and health care. PMID- 7495446 TI - Teaching crisis intervention skills to medical students. PMID- 7495447 TI - Do housestaff interview standardized patients differently from real ones? PMID- 7495448 TI - Validation of standardized-patient assessment: a meaning for clinical competence. PMID- 7495449 TI - The outlook for clinical research: impacts of federal funding restraint and private sector reconfiguration. AB - Accustomed to congressional and industry support, patient-centered clinical research is at a crossroads in 1995. Forced to look into the next century by the seven-year budget cycle selected by Congress, its path seems hindered by threatened cuts in funding for the National Institutes of Health (NIH), cost pressures on private-sector organizations that support research initiatives, and market restraints on the academic health centers that traditionally have served as research bases for many clinical investigators. Assuring public and private financial support for clinical research has become an important topic on policy and industry agendas, as has the need to sort out the roles of the NIH, academic health centers, managed-care plans, and pharmaceutical firms in backing and conducting research. The very character of clinical inquiry seems to be reshaping itself amid conflicts over basic versus applied research, ongoing versus new investigations, and scientific discovery versus product development. How these issues are approached--let alone resolved--is significant not only for the future of clinical research but also for the health of the public. The author discusses these issues and concludes with a list of specific questions that must be addressed in confronting policy issues of clinical research. PMID- 7495450 TI - Realigning the values of academic health centers: the role of innovative faculty management. AB - As a market economy continues to permeate U.S. health care, fiscal accountability imposed by purchasers of medical services will reduce funds for medical education and will heighten the scrutiny of that activity. The authors propose that U.S. academic health centers must respond to these changes in the health care environment by critically examining their culture and values. This process will call for a reorientation of their values to place clinical education at the center of the academic enterprise. The authors challenge the notion that research and publications-oriented faculty are the best group to train optimally effective clinical practitioners. They argue that although faculty promotion and rank in American medical schools are highly correlated with publications and funded research, there is no body of evidence that shows that fecundity in research or publications is essential for faculty to be excellent medical educators. The authors maintain that the main tool for realigning the fundamental values of academic health centers will be a new form of faculty management by the medical leadership. After outlining the current approach to research, teaching, clinical service, and administration at academic health centers, the authors challenge the traditional view that a faculty member should master all of these four elements, and state that the era of the "quadruple-threat" faculty member is passing. What can emerge is an emphasis on departments' and institutions' becoming "quadruple threats," which can occur only if institutions' leaders become better managers of their individual faculty members' priorities.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7495451 TI - A promotion ladder for teachers at Harvard Medical School: experience and challenges. AB - The authors describe the development, implementation, and institutionalization at Harvard Medical School of a promotion ladder that recognizes the teaching and scholarly contributions of full-time clinical faculty. They also discuss the challenges that arose during this process, for example, how to make the new track creditable and attractive to both the appointed faculty and the faculty at large. The criteria developed for promotion focus on a candidate's skills and accomplishments in teaching, scholarship, clinical work, and departmental service. The authors present the elements of these criteria for the three professional levels of the ladder and outline the appointment process, including the steps for consideration of a given promotion. The development of this teacher clinician ladder has had a positive influence on faculty who are committed to teaching by allowing recognition of their contributions in a track held to be the equal of the other full-time tracks in a medical faculty traditionally committed to research and patient care. Data are given for the 70 faculty who were promoted over the five years from 1989 to 1994. The true success of this promotion ladder will be measured only over time through its impact on the educational enterprise within the medical school and its hospitals, and its capacity to both successfully advance the careers of qualified medical educators and further the development of the field of medical education. PMID- 7495452 TI - Revising the definition of the generalist physician. AB - Although there has been growing recognition of the important role of generalist physicians in the United States, there has been insufficient discussion of who the generalists are and how their role is defined. Traditionally, generalists have been defined by their specialty, with physicians in family medicine, general internal medicine, and general pediatrics considered generalists. But this approach may not sufficiently recognize the specific competencies and therefore the specific training required of generalists. A new definition is based on the functional requirements of generalist practice and the central role that generalist physicians will play in comprehensive care. This definition can be used, first, to assure that medical education, both fundamental and continuing, provides generalists with these needed competencies and, second, to judge claims among different categories of physicians that they should be considered generalists. PMID- 7495453 TI - Doctoring III: innovations in education in the clinical years. AB - Despite recent major changes in the practice of medicine, there has been relatively little change in medical education, particularly in the clinical years. Important areas such as ethics, domestic violence, nutrition, preventive medicine, and clinical decision making have been neglected in the curriculum. However, in 1994 the UCLA School of Medicine began to implement Doctoring III, a multidisciplinary, centralized, longitudinal course that spans the third year of medical school. This course addresses many underrepresented topics. Students spend one day every other week in Doctoring III. Half of the day is spent in community-based clinical sites and the other half is spent in small-group teaching sessions. In the small groups, students and faculty follow and manage a panel of simulated patients over the course of the year. The students thus have the opportunity to develop a sense of the progression of common illnesses over time and to explore related ethical, social, and other concerns. The approach taken in Doctoring III has enabled the UCLA School of Medicine to overcome many barriers to curricular change, and it may serve as a model for incorporating the teaching of underrepresented topics in the clinical years. PMID- 7495455 TI - Integrating behavioral and social sciences research at the NIH. PMID- 7495454 TI - Issues and challenges in the design of curriculum information systems. AB - Medical schools are increasingly using curriculum information systems to better manage their curricula, to incorporate alternative learning environments, to expand subject-specific instruction, or to adapt the curricula to the changing health care environment. A curriculum information system contains key features, selected by the system's designers, that describe the objectives, the specific content, and/or the educational activities that compose the curriculum. The underlying purpose of such a system is to document and describe the knowledge, behaviors, skills, attitudes, or activities students will be expected to develop or learn. While it might be ideal to have one system that would meet the needs of all possible users, the programming and maintenance requirements of such a single system would exceed most medical schools' resources. Thus, designers of curriculum information systems must first identify the primary intended-user group: students, faculty, or administrators. The system designed for one group will typically differ from the systems for other groups in the emphasis on and level of content detail and curriculum structure information. The general structure and purposes of curriculum information systems are expressed in three distinct system designs: a curriculum database (most helpful for administrators), a curriculum textbase (for faculty), and an electronic syllabus (for students). The authors describe these different designs, and they discuss challenges faced by system designers. PMID- 7495456 TI - Professional networks: the influence of colleagues on the academic success of faculty. AB - BACKGROUND: Successful higher education faculty, those who get promoted and tenured, who get recognized for contributions, who produce more and significant research, frequently consult colleagues. This article summarizes what is known about colleague relationships with the hope of stimulating further research to extend current conclusions to medical school faculty. METHOD: In the spring of 1992, a systematic literature review was conducted using database searches and author review of 137 selected books and articles using a standard protocol; preference was given to articles that were data-oriented, used quality designs, and related directly to the topic. RESULTS: Forty-seven "best" sources, published between 1963 and 1991, were included in the review of (1) types and configurations of colleague relationships, (2) forming and maintaining colleague relationships, (3) colleague effects on faculty success, (4) functions of colleague relationships, (5) changes in colleague relationships over a career, and (6) effects on methods, size, and configuration of colleague relationships. CONCLUSION: Among the conclusions reached are that (1) there are a variety of types or configurations of colleague relationships, all with different functions and effects on faculty performance; (2) dyadic conceptions of colleague relationships are insufficient to explain the functions of colleague relationships; and (3) the most important source for developing colleague relationships is professional associations, while the least important source is one's own institution. PMID- 7495457 TI - Withdrawal and extended leave during residency training: results of a national survey. AB - BACKGROUND: Although national figures for medical student withdrawal and extended leave have long been reported, similar data have not been available for residents in training. METHOD: Data for this study came from the American Medical Association survey of the 1991-92 residency year, in which program directors were asked for information about residents who had taken extended leave or had withdrawn or been dismissed from their programs prior to completion. Data are reported for 89,368 residents enrolled in 6,302 programs (89.2% of all surveyed programs). RESULTS: During the 1991-92 year, 2,449 residents (2.7%) withdrew or were dismissed from their programs and 887 (1.0%) took extended leave. Specialty and program changes accounted for 56% of the withdrawals, while performance difficulties were implicated in 12.9%. Maternity or paternity leave was involved in 32.2% of extended leaves, followed by research sabbaticals (11.4%) and physical problems (10.5%). Women had higher rates of both withdrawal and extended leave than men. Withdrawal for performance difficulties was lowest among graduates of U.S. and Canadian allopathic schools as compared with graduates of osteopathic or foreign medical schools, and lowest among Caucasians as compared with those of other racial-ethnic identities. CONCLUSION: Although overall figures and percentages are low, there are small but persistent losses of residents annually that vary by specialty, gender, race-ethnicity, and education. PMID- 7495458 TI - Should half of all medical school graduates enter primary care? Perceptions of faculty members at Jefferson Medical College. AB - PURPOSE: This study was undertaken to promote communication among faculty regarding the impact of a proposed goal that 50% of the graduates of Jefferson Medical College enter generalist careers. Since the opinions and attitudes of faculty regarding career decisions may directly or indirectly influence students, the authors investigated faculty's views of the optimal ratio of primary care to non-primary care physicians in the workforce and their perceptions of the effect on medical education, research, and health care delivery if the 50% goal were to be mandated. METHOD: A questionnaire was mailed in January 1994 to all 684 salaried faculty of Jefferson Medical College. Respondents' opinions about the optimal primary care to non-primary care ratio and their perceptions of the effects of implementing the 50% goal on 21 areas related to medical education, research, and health care delivery were examined using a Likert-type scale. Obstacles perceived by non-primary care physicians as preventing their practice of primary care were also among the outcome measures. RESULTS: A total of 275 completed questionnaires were received (40% response rate; 72 primary care physicians, 141 non-primary care physicians, and 62 non-physicians). The median and mode of an optimal primary care to non-primary care ratio were both 50/50. Faculty, in general, perceived that implementing the 50% goal would enhance public access to primary care, physician-patient relationships, utilization of non-physicians, and the career satisfaction of generalists. They predicted decreases in costs of care, freedom of career choice, funding, and interest in research. The primary care physicians perceived greater enhancements of the image of physicians, quality of care, and satisfaction of generalists and subspecialists than did the non-primary care physicians. Gender and age did not affect the perceptions. A lack of appropriate training was identified by 45% and a lack of interest by 28% of the non-primary care physicians as major obstacles to their practice of primary care medicine. CONCLUSION: The faculty members' positive and negative views of the proposed reform can provide useful information to the institution in understanding the potential impediments to increasing the numbers of generalist graduates. The generalists had significantly different views from the subspecialists about the impact of increasing the proportion of primary care physicians on health care delivery and research. In general the primary care physicians were more likely to view the proposed changes as beneficial than were the non-primary care physicians. PMID- 7495460 TI - Telephone management training in internal medicine residencies: a national survey of program directors. AB - BACKGROUND: Little is known about how internal medicine residents train for and practice telephone management. To address this deficiency, a national survey of program directors at accredited internal medicine training sites was conducted to evaluate residents' training for and practice of telephone medicine. METHOD: A 43 item questionnaire was mailed in December 1993 to all program directors at the 416 accredited internal medicine training sites in the United States. A limited questionnaire, regarding the most essential training questions, was mailed to all non-responders. RESULTS: The response rate was 60% (250) for the full questionnaire. Only 15 (6%) of the programs offered formal training in telephone management to their residents. This training usually consisted of single lectures (nine programs) or reading materials (seven programs). The respondents felt that formal training in telephone management was very important (155, 62%) and that such training should be a part of every internal medicine curriculum (150, 60%). CONCLUSION: Few internal medicine programs offered training in telephone management. When training occurred, it was usually limited and informal. Most program directors felt that training was important and that current training efforts were unsatisfactory, emphasizing the need for curriculum development and implementation in telephone management. PMID- 7495459 TI - A model for predicting HEAL repayment patterns and its implications for medical student financing. AB - BACKGROUND: The current level of Health Education Assistance Loan (HEAL) defaults has exceeded the original estimate, and as such is producing an unforeseen federal expenditure. Predicting repayment patterns of HEAL borrowers is an important step in assessing the impact that this unforeseen expenditure will have on HEAL and other financial aid programs. METHOD: Since prior research on educational lending has not identified a practical model for predicting repayment patterns, the authors turned in 1994 to an alternative discipline, research on consumer lending. Using the multivariate discriminant analysis credit scoring model framework, the authors incorporated operational definitions of the borrower's character, capacity, and capital. To identify factors that are significant in establishing repayment category membership, the framework was applied to a group of 233 HEAL borrowers who graduated from five medical schools in 1988. Because of the small numbers of delinquent borrowers and defaulters, the repayment categories were restricted to repayment, deferment, and forbearance. RESULTS: The level of unsubsidized debt, in conjunction with financial resources (including the potential resource of parental support), may be significant in identifying those HEAL borrowers who may confront repayment difficulty. CONCLUSION: While the results are not surprising, they do lend documented support for a review of financial aid policies at both institutional and governmental levels, and a framework within which this review may be conducted. PMID- 7495461 TI - The relationship between parental income and academic performance of medical students. AB - PURPOSE: To test the hypothesis that family financial status is associated with the academic performance of a medical student. METHOD: The relationships between parental income and mean scores on the Medical College Admission Test (MCAT) and United States Medical Licensing Examination (USMLE) Step 1 were examined for the students in the 1994 and 1995 graduating classes at the UMDNJ-New Jersey Medical School who had applied for financial aid in 1991 and reported annual parental income. Pearson correlations were used to analyze separately the data for minority and majority students, for men and women, and for the four subgroups by gender and race-ethnicity. RESULTS: The final study cohort consisted of 192 students (55% of all students). Significant positive correlations were found between the (1) MCAT and USMLE Step 1 for the women, men, majority, and minority students, (2) MCAT and parental income for the subgroups of majority men and minority women, and (3) USMLE Step 1 and parental income for the subgroup of minority women. CONCLUSION: Parental income was correlated significantly with performances on the MCAT and USMLE Step 1. These relationships may be particularly strong and persistent for minority women. PMID- 7495462 TI - Comparing the admission interview ratings and comments of faculty and student interviewers. PMID- 7495463 TI - Effect of learning objectives on the performances of students and interns rotating through an emergency department. PMID- 7495464 TI - Use of student feedback by curriculum decision makers and its relation to the quality of their teaching. PMID- 7495465 TI - Influence of tutors on students' perceptions of small-group, case-based courses. PMID- 7495466 TI - Specialty intentions of 1995 U.S. medical school graduates and patterns of generalist career choice and decision making. AB - The authors report on the specialty intentions that graduating students declared on the 1995 AAMC Medical School Graduation Questionnaire (GQ) and compare the pattern of career choices in 1995 with that in 1992. Family practice was the leading choice of graduates in 1995, followed by internal medicine subspecialties and general internal medicine. These choices represented significant gains over those made in these specialties in 1992 and were at the expense of declines in the interest of 1995 graduates for internal medicine specialties, radiology, anesthesiology, obstetrics-gynecology subspecialties, and some other fields. In 1992, 14.6% of graduating students declared plans to pursue careers in one of the generalist specialties; in 1995, 27.6% declared such plans. In 1992, no school graduated 50% or more students with generalist intentions, and only one school reached 40%; in 1995, five schools graduated more than 50%, and another 15 graduated more than 40% who favored generalist careers. Medical schools with significant GQ response rates (110 out of 125) were aggregated by level of generalist production (top 25%, middle 50%, and bottom 25%) according to the percentages of their 1995 graduates selecting careers in the individual generalist specialties of family practice, general internal medicine, and general pediatrics, and in these generalist specialties in toto. Within these groups, the linking of GQ responses to declarations given by the same students on the Matriculating Student Questionnaire (MSQ) made it possible to determine the extent to which graduates' specialty choices represented early interests that were retained or interests acquired later during medical school.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7495467 TI - Pentatomic cyclic agonists and muscarinic receptors: a 20 years review. AB - The results of 20 years of structure-activity relationship (SAR) studies of pentatomic cyclic agonists on muscarinic receptors are reported and commented. Our results are compared with the findings obtained from quantitative structure activity relationship (QSAR) studies and from those of molecular biology. The results from all three approaches point in the same direction and should help in the future design of new muscarinic agonists. PMID- 7495468 TI - Structure-activity relationships in open ansa-chain rifamycin S derivatives as inhibitors of HIV-1 reverse transcriptase. AB - Three types of open ansa-chain rifamycin S derivatives have been prepared: derivatives with the ansa-chain open at C(29) and the original dihydrofuranone ring; derivatives with the ansa-chain open at C(29) and a furane ring; derivatives with the ansa-chain at open NH-C(15). Only derivatives of the first type are weak inhibitors of HIV-1 reverse transcriptase (IC50 ca.300 microM) while derivatives of the two other types are inactive. It has been hypothesized that the active derivatives inhibit the viral enzyme interacting through the groups C(14)H3, C(13)H3, and C(1)O at the same site as the well-known inhibitors TIBO and Nevirapine. In particular C(13)H3 must be unhindered and in an appropriate position out of the plane containing the chromophore-rings. The open ansa-chain seems to play the role of a lipophylic substituent. PMID- 7495469 TI - Biological effects of hesperidin, a Citrus flavonoid. (note II): hypolipidemic activity on experimental hypercholesterolemia in rat. AB - Hesperidin, the most important flavanone of Citrus sp., significantly increases HDL and lowers cholesterol, LDL, total lipid and triglyceride plasma levels in normolipidemic rats and in rats with diet- and triton-induced hyperlipidemia. PMID- 7495470 TI - Inhibitory properties of triethylphosphine goldlupinylsulfide in adjuvant-induced arthritis in rats. AB - A new gold coordination compound (triethylphosphine goldlupinylsulfide: TP-Au-LS) was tested in adjuvant-induced arthritis in the rat, by oral administration at doses of 5, 10 and 20 mg/kg/day of gold for 17 consecutive days, in comparison with auranofin and betamethasone. TP-Au-LS produced a dose dependent reduction of both the injected and uninjected hind paw volume. Gold levels in serum (measured on day 18 by inductively coupled plasma atomic emission spectrometry) were also found to be dose related. At the dose of 10 mg/kg, TP-Au-LS and auranofin induced superimposable reductions of the injected paw volume; however the first drug produced higher serum gold concentrations than those achieved with the latter. PMID- 7495471 TI - Synthesis and pharmacological properties of thieno[2',3':4,5]pyrimido [2,1 b][1,3,4]thiadiazine derivatives. AB - A new series of 2-substituted-7,8-dimethyl-3H,9H-thieno[2',3':4,5]pyrimido[2,1- b][1,3,4]thiadiazin-9-ones 5a-d and 6a,b was synthesized through the hydrazinium(1+) salt of 3-amino-2,3-dihydro-5,6-dimethyl-2-thioxo- thieno[2,3 d]pyrimidin-4(1H)-one, 2. These derivatives and two analogs 10a,b were tested for their analgesic and antiinflammatory properties. The pharmacological results are discussed in comparison with those of related compounds previously tested. PMID- 7495472 TI - Synthesis and antimicrobial activity of some tetramethylenethieno[2,3 d]pyrimidine derivatives. AB - A series of tetramethylenethieno[2,3-d]pyrimidine derivatives has been synthesized and tested for its antimicrobial properties. All the synthesized compounds were found to exhibit in vitro antibacterial and/or antifungal activity. The highest activity was elicited by 4-benzolhydrazino-5,6 tetramethylenethieno[2,3-d]pyrimidine (9) showing MIC value of 7.81 micrograms/ml against E. Coli and C. albicans, while its MBC value was half that of nystatin. Compound 16 was almost as potent as nystatin exhibiting a minimum bactericidal concentration (MBC) value of 15.62 micrograms/ml. PMID- 7495475 TI - Evaluation of polysaccharides intended for ophthalmic use in ocular dosage forms. AB - Different water-soluble polysaccharides were evaluated for their intended use in ocular dosage forms. The physicochemical characteristics and the viscosity of the solutions prepared with several iso-osmotic vehicles were measured. The influence of ions present in lacrimal fluid, mucin and cyclodextrin on the rheological behaviour was examined. Scleroglucan and xanthan gum exhibit viscoelastic and mucoadhesive properties useful for ocular dosage form formulations. PMID- 7495473 TI - Research on antibacterial and antifungal agents. XII--Synthesis and antimicrobial activity of some Mannich bases of diarylpyrroles. AB - The synthesis and antimicrobial activity of some Mannich bases of diarylpyrroles are reported. The obtained data show that activity is closely connected to molecular basicity. Results are discussed on the basis of structure activity relationships. PMID- 7495474 TI - Synthesis and biological activity of novel metal complexes of 2-acetylpyridine thiosemicarbazones. AB - 2-Acetylpyridine-(2-methylthiosemicarbazone), 2-acetylpyridine-(4 methylthiosemicarbazone), 2-acetylpyridine-(4-phenylthiosemicarbazone) and some of their metal complexes of the platinum group have been synthesized, characterized by chemical and spectral methods and studied for their antibacterial, antifungal and amoebicidal activity in vitro. They were studied also for their antimalarial activity and for toxicity in vivo. The platinum metal chelates exhibited significant activity against a wide spectrum of microorganisms at different concentrations. The Ru (III) chelates derived from 2-acetylpyridine (4-methylthiosemicarbazone) seem to be the most efficient inhibitors. Evaluation of the antimalarial activity of the complexes in mice infected with Plasmodium berghei indicated that cures were attainable at dose levels of 20-160 mg/kg. PMID- 7495476 TI - Identification of anthracenyl-dipeptide conjugates as novel topoisomerase I and II inhibitors and their evaluation as potential anticancer drugs. AB - As part of an ongoing rational drug design programme aiming to develop monosubstituted anthracenyl-peptides as potential anticancer drugs, three novel dipeptide conjugates have been synthesized and evaluated as inhibitors of topoisomerase (topo) I and II. Each of the three conjugates (designated NU/ICRF 600-602) was shown to inhibit the catalytic activity of both topoisomerase I and II, of which NU/ICRF 602 was the most active [100% inhibition of both enzymes at 5 micrograms/ml (approximately 15 microM) or less]. In a topo I/DNA unwinding assay, none of the compounds bound to DNA, suggesting genuine inhibition of catalytic activity. NU/ICRF 600 stabilized topo I cleavable complexes, although none of the compounds induced topo II-mediated DNA cleavage. Using a panel of Chinese hamster ovary cell lines along with the human ovarian cancer cell line, A2780, none of the three compounds were actively cytotoxic at concentrations < 100 microM. Subsequent drug uptake studies with NU/ICRF 600 and 602, using a method developed to correlate the chemosensitivity of A2780 cells with the uptake of anthracenyl-amino acid conjugates, revealed a lack of cellular uptake for both dipeptide conjugates. The significance of this finding in relation to drug design and the future development of this series of compounds is discussed. PMID- 7495477 TI - Antineoplastic agents 337. Synthesis of dolastatin 10 structural modifications. AB - New structural modifications of the marine shell-less mollusk peptide constituent dolastatin 10 (1) have been synthesized, and evaluated against a variety of cancer cell lines and for their ability to inhibit tubulin polymerization. A number of useful structure-activity relationships were uncovered. The most important observation was that the dolaphenine unit of dolastatin 10 could be satisfactorily replaced with a phenethylamine. Peptide 11C, designated auristatin PE, was found to exhibit inhibition of cancer cell growth and tubulin assembly comparable to that of dolastatin 10. PMID- 7495478 TI - Synthesis of podophyllotoxin congeners as potential DNA topoisomerase II inhibitors. AB - We have synthesized a number of new podophyllotoxin congeners in which the A-ring is opened and the C-ring is aromatic. These are based on the chemically modified structures of the naturally occurring podophyllum lignan, diphyllin. These have been synthesized by employing the Michael Initiated Ring Closure methodology. These lignans exhibited considerable DNA topoisomerase II inhibition and were devoid of topoisomerase I inhibition; they did not exhibit very significant activities in the in vitro human tumour cell lines assay. The results obtained provide insights into the structure-activity relationships and the design of chemically modified podophyllotoxin congeners useful in cancer chemotherapy. PMID- 7495479 TI - Quinazoline thymidylate synthase inhibitors: methods for assessing the contribution of polyglutamation to their in vitro activity. AB - Many quinazoline thymidylate synthase (TS) inhibitors undergo intracellular metabolism to polyglutamate forms which can significantly alter their activity and pharmacodynamics through improved TS inhibition and drug retention. When a series of quinazolines was tested for inhibitory activity towards TS (IC50 0.001 2 microM) and the growth of L1210 cells (IC50 0.005-10 microM), no direct correlation was observed. However, a very good correlation was apparent if a L1210 variant cell line (L1210: RD1694) was used. This line is deficient in its ability to form antifolate polyglutamates. A number of other intact cell methods have also been developed which estimate the contribution that intracellular polyglutamation makes to a compound's activity. These assays were validated using a series of quinazoline-based TS inhibitors with well-defined activity for TS, folypolyglutamate synthetase (FPGS) and the reduced-folate cell membrane carrier (RFC). Short-exposure growth-inhibition assays or the measurement of TS activity in situ after various incubation times, followed by different lengths of time in drug-free medium, can indicate both the speed and extent of appearance of retentive forms (usually polyglutamates). Continuous-exposure growth-inhibition assays, in the presence of leucovorin (LV), are also useful, since only the growth-inhibitory potency of polyglutamated analogues is significantly decreased by LV. Highly polyglutamated compounds, e.g. ZD1694, are virtually inactive in the presence of a high concentration of LV. It is proposed that these methods, when considered together, provide a greater degree of information concerning the rate and extent of polyglutamation of a particular compound than isolated FPGS assays alone. PMID- 7495480 TI - Quinazoline-based thymidylate synthase inhibitors: relationship between structural modifications and polyglutamation. AB - Quinazoline-based analogues of folic acid are a group of thymidylate synthase (TS) inhibitors that display a wide spectrum of activity for cultured tumour cells, partly due to their differential ability to form polyglutamate metabolites that are (i) more potent TS inhibitors and (ii) not readily effluxed from cells. The rate of cell membrane transport and folylpolyglutamate synthetase substrate activity influence compound polyglutamation. A series of intact-cell assays has been used to determine how specific modifications of 2-desamino-2-methyl-N10 propargyl-5,8-dideazafolate (ICI 198583) affect compound polyglutamation. Those containing the 'classical' glutamate structure were usually, but not always, well polyglutamated intracellularly. Replacement of N10 propargyl with smaller aliphatic substituents, particularly when combined with replacement of the benzene ring with thiophene or thiazole heterocycles, was beneficial for antitumour activity through polyglutamate formation. Fluorination of the benzene, particularly if a F was adjacent to the 'bridge region' (3'F or 2',5'diF), also gave compounds with a high dependence on polyglutamation for activity. Those analogues with 2-CH2OH or NH2 substituents were poor substrates for the reduced folate cell membrane carrier which can account for their reduced polyglutamation rate and hence growth-inhibitory activity. A large decrease or prevention of polyglutamation was achieved by the introduction of CH3, CH2CH3, Br or C1 on C7. The concomitant enhancement in TS inhibition by these modifications gave compounds active under continuous-exposure cell culture conditions. Some ICI 198583 analogues had the glutamate moiety replaced with unnatural amino acids or dipeptides. Only the L-gamma-L-glu analogue (a polyglutamate metabolite of ICI 198583) gave activity entirely attributable to polyglutamate formation. PMID- 7495481 TI - Heteronuclear NMR pulse sequences applied to biomolecules. AB - Current concepts in heteronuclear multidimensional NMR spectroscopy are reviewed. Methods to improve the sensitivity and the efficiency of data collection include constant time, compression through the overlap of chemical shift evolution and dephasing and rephasing periods, and dual or time-shared evolution. Two classes of three-dimensional and four-dimensional triple-resonance experiments applied to proteins are considered. The first class correlates 1H, 15N, and 13C signals of the protein backbone. The second class correlates both backbone and side-chain signals. Application of triple resonance to RNA is also discussed. Heteronuclear cross polarization (HCP) is considered as an alternative to INEPT transfer, and its application to nucleic acids is presented. Finally, two methods of employing pulsed field gradients (PFGs) are reviewed. PMID- 7495482 TI - Salt-nucleic acid interactions. AB - Coulombic interactions of salt ions with polymeric and oligomeric nucleic acids in solution have large and distinctive effects on ion distributions, on thermodynamic coefficients, and hence on equilibrium processes involving nucleic acids, such as their conformational transitions and binding interactions. In experimental or theoretical studies where an oligonucleotide is taken to represent the corresponding polynucleotide, the impact of coulombic end effects on molecular and thermodynamic properties must be taken into account. Observable consequences of coulombic interactions in nucleic acid solutions have been calculated by using models with varying degrees of detail and methods formulated at varying levels of rigor. From comparisons of experimental results with predictions of the prevalent theoretical approaches, this review concludes that the more rigorous methods have proved capable of accounting for thermodynamic (and some molecular) consequences of coulombic interactions with a minimal number of preaveraged parameters that represent the most important structural features of the nucleic acid solution. PMID- 7495483 TI - Young people and alcohol, drugs and tobacco. PMID- 7495484 TI - Allergy: who, why and what to do about it? PMID- 7495485 TI - The cell biology of phagocytes. PMID- 7495486 TI - Immunodeficiencies of genetic origin. PMID- 7495488 TI - Langerhans cells and HIV infection. AB - Epidermal Langerhans cells (LCs) isolated from individuals infected with human immunodeficiency virus (HIV-1) harbour HIV-1 proviral DNA and RNA, indicating productive infection by the virus in vivo. Furthermore, normal LCs can be infected in vitro by HIV and can present HIV antigens to helper T cells. Here, Giovanna Zambruno and colleagues discuss the possibility that LCs of genital mucosae are among the first targets of HIV infection following sexual contact, and can be involved both in the transmission of the infection to T cells and in T cell priming to HIV antigens. In addition, epidermal LCs might acquire HIV infection from dermal T cells during transit from blood vessels through the dermis and may, in turn, represent a reservoir of the virus for continued T-cell infection. PMID- 7495487 TI - AIDS: a disease of impaired Th-cell renewal? AB - Individuals who are infected with human immunodeficiency virus (HIV) and develop AIDS are characterized by the progressive loss of T helper (Th) cells, together with an increase in virus load. Despite a remarkable similarity to humans, and their susceptibility to persistent HIV-1 infection, chimpanzees have a relative resistance to the development of AIDS. Here, Jonathan Heeney proposes that the critical underlying event perpetuating the progression to AIDS is the impairment of the immunological microenvironment necessary for competent, antigen-specific, Th-cell renewal. Survival is dependent on the host's ability to preserve the immunological infrastructure sufficiently in order to maintain the capacity for renewal of a balanced, competent Th-cell population. PMID- 7495489 TI - The P2Z purinoceptor: an intriguing role in immunity, inflammation and cell death. AB - Many immune and inflammatory cells express a plasma membrane receptor for extracellular ATP, termed the P2Z purinoceptor, which appears to be coupled to a plasma membrane pore. The physiological role of such a molecule is generally unknown, except for the striking susceptibility to ATP-mediated cytotoxicity that it confers. The receptor is upregulated in human monocytes by interferon gamma and is also expressed during macrophage differentiation. Here, Francesco Di Virgilio discusses recent information on this receptor, and suggests a possible role for it in the immune and inflammatory response. PMID- 7495490 TI - The human immune system in hu-PBL-SCID mice. AB - Severe combined immunodeficiency (SCID) mice can be stably grafted with human peripheral blood lymphocytes, creating hu-PBL-SCID chimeras; essentially, these are mice with a human immune system. Here, Magdalena Tary-Lehmann, Andrew Saxon and Paul Lehmann discuss the immunobiology of these chimeras. The authors propose that hu-PBL-SCID chimerism evolves in two phases. During the first three weeks after grafting, many of the injected cells survive and the human immune system is functional. Subsequently, anti-mouse-reactive clones are selected and the immune system becomes nonfunctional. The implications of this scenario for the utilization of the hu-PBL-SCID model are discussed. PMID- 7495491 TI - Involvement of B-1 cells in mucosal immunity and autoimmunity. AB - B-1 cells are distinguished from conventional B cells by their anatomical localization, surface phenotypes and functional characteristics. The physiological functions and pathological roles of these cells remain controversial. In this review, Masao Murakami and Tasuku Honjo summarize recent evidence for the involvement of B-1 cells in mucosal immunity and autoimmunity, and discuss the relationship between these phenomena. PMID- 7495492 TI - Molecular defects in the bare lymphocyte syndrome and regulation of MHC class II genes. AB - The complex pattern of expression of major histocompatibility complex (MHC) class II molecules plays an essential role in the control of the immune response. Our understanding of the molecular mechanisms controlling this expression has benefited greatly from the identification of the regulatory factors defective in two forms of a hereditary disease of MHC class II regulation: bare lymphocyte syndrome. This has also provided new tools for the experimental modulation of MHC class II expression. PMID- 7495493 TI - Negative selection for natural killer cells? PMID- 7495494 TI - Dendritic cells as initiators of tumour immunity. PMID- 7495495 TI - Signaling by HLA class II antigens on B cells. PMID- 7495496 TI - Immunology of centenarians. PMID- 7495497 TI - P-glycoprotein: clinical significance and methods of analysis. AB - Multidrug resistance (MDR) is responsible for a decrease in sensitivity of tumor cells tumor cells to unrelated, naturally occurring anticancer drugs. This resistance is correlated with expression and activity of a membrane protein, P-gp 170, functioning as a drug-extruding pump. It has been well described in in vitro situations; however, the clinical detection and implications are not yet clear. Multiple detection assays have been developed based on the discovery of the MDR gene family and the corresponding protein. Southern, Northern, or Western blot analysis, S1 nuclease protection or PCR-based assays, immunohistochemical detection or functionality tests by flow cytometry have been used extensively. However, by use of these techniques on clinical material, both normal and malignant, contradictory results have emerged. The sensitivity and specificity of a certain technique are always limited by unavoidable parameters, for example, skill of the technician. Moreover, the complexity of the development of resistance against anticancer agents (external determinants), such as the diversity of tumor tissues, the simultaneous presence of other resistance mechanisms, and the low expression level, make MDR detection equivocal and can lead to contradictory results. Previous treatment influencing the MDR profile and inappropriate timing of the test make a possible correlation between MDR expression and chemotherapeutic resistance difficult to establish and can lead to discordant results. In this review, the need for proper criteria is stressed. No single detection technique provides the ideal test to detect MDR. Tandem testing could give more certainty, although small sample size limit this application. Formulation of a standard assay with better definition of a positivity is essential before clinical trials are started. PMID- 7495498 TI - Physiology and pathophysiology of complement: progress and trends. AB - The complement system comprises a family of at least 20 plasma and membrane proteins that interact in a tightly regulated cascade system to destroy invading bacteria and prevent the deposition of immune complexes in the tissues. This brief review addresses the basic mechanisms of complement activation and control and describes the active fragments produced during complement activation. The biological importance of the complement system is amply illustrated in patients with complement deficiencies, who are susceptible to bacterial infections and immune complex diseases. The involvement of complement in other immunological diseases is an expanding area of clinical research, supported by the development of new assays for the identification of complement activation. This area is discussed here with particular reference to neurological diseases. A promising new prospect involves the use of complement inhibitory molecules in therapy of complement-mediated disease and this exciting area is also discussed. Novel physiological roles of complement also are being revealed and new evidence that complement and complement receptors play an important role in reproduction is summarized. It is hoped that this brief overview will convey some of the enthusiasm currently pervading research in this underappreciated area of immunology. PMID- 7495499 TI - Parathyroid hormone-related protein. AB - We review the current state of knowledge of the molecular properties and actions of parathyroid hormone-related protein (PTHrP) both in cancer patients and in normal physiology. PTHrP is a common product of squamous cancers and is the major mediator of the syndrome of humoral hypercalcemia of malignancy (HHM) by its actions through parathyroid hormone receptors in bone and kidney. Recently developed radioimmunoassays and tissue localization techniques indicate that PTHrP is produced by many more cancers than was originally indicated by clinical studies and that it contributes significantly to malignancy-related hypercalcemia associated with other etiologies, for example, cancers metastatic to bone and hematological malignancies. The gene encoding PTHrP is complex, with multiple exons coding for up to 12 alternate transcripts and three different length proteins, potentially in a tissue-specific manner, by the use of three promoters. Its expression is regulated by hormones and growth factors, and the untranslated exons display features in common with many cytokine genes. Although potential endocrine actions of PTHrP are evident in fetal development, further evidence suggesting that the normal physiological role of PTHrP is predominantly as a locally produced regulator/cytokine comes from localization studies and investigations of its actions in a variety of tissues. Such studies indicate that in addition to its parathyroid hormone-like actions, PTHrP has multiple activities, including those in fetal development, placental calcium transfer, lactation, smooth muscle relaxation, and on epithelial cell growth. Although PTHrP was discovered because of its production by cancers, evidence for its actions as a local regulator highlights the importance of understanding its roles not only in the etiology of HHM in cancer patients but also in normal tissues. PMID- 7495500 TI - Analgesic rebound headache. How great is the problem and what can be done? PMID- 7495501 TI - Corticosteroids in kidney transplant recipients. Safety issues and timing of discontinuation. AB - Corticosteroids have played a key role in the immunosuppression of organ transplantation. Unfortunately, the extensive use of these agents has resulted in disabling and life-threatening adverse effects in many patients. The advent of concomitant corticosteroid/cyclosporin regimens has allowed a reduction in the dosages of steroids administered, yet steroid-induced morbidity is still a major problem in many cyclosporin-treated renal transplant patients. After favourable initial experiences with cyclosporin monotherapy, several attempts at steroid free immunosuppression in renal transplant patients have been undertaken, either by not starting steroids after transplantation or by stopping steroids in patients with stable graft function. Most controlled and uncontrolled trials showed that with either strategy short term graft survival was similar with or without steroids, but acute rejection was more frequent in patients not taking steroids. The percentage of patients who could be maintained steroid-free ranged from 28 to 94%, and was higher in patients who stopped steroids later than in those never receiving them. Little information is available about long term follow-up of these patients. Some studies reported late attrition of renal function in patients not taking steroids, while others reported a favourable outcome even in the long term. Steroid-free immunosuppression is feasible in renal transplant patients, but it requires careful monitoring of renal function and cyclosporin dosage. This strategy is particularly indicated in patients at high risk of cardiovascular disease or steroid-related complications, and in children. Nevertheless, several issues need to be better elucidated by further studies, namely the long term outcome of steroid-free immunosuppression, the advantages and disadvantages of steroid avoidance versus steroid withdrawal, and the criteria for selecting patients. PMID- 7495504 TI - Immunodeficiency. PMID- 7495505 TI - Potential targets for autosomal SCID mutations. AB - Significant advances have been recently made in the molecular characterization of genes that encode proteins with activities that are directly, or indirectly, involved in the assembly of antigen receptor variable region genes. Such genes are candidate targets for human autosomal mutations that lead to severe combined immune deficiencies characterized by a lack of both T and B cells. PMID- 7495503 TI - The risks and benefits of antimicrobial therapy in pregnancy. AB - Antimicrobial agents, especially antibiotics, are prescribed in pregnancy for various specific indications related to pregnancy per se (e.g. chorioamnionitis), for infections otherwise unrelated to but complicating pregnancy (e.g. pneumonia, bacterial endocarditis) and prophylactically for conditions which if untreated would have an adverse outcome for the pregnancy (e.g. asymptomatic bacteriuria). The selection of an agent will depend on the likely pathogen, stage of pregnancy, special pharmacokinetic factors related to the stage of pregnancy, safety of the drug and cost. Some classes of antibiotics can be taken throughout the 3 trimesters (e.g. beta-lactams), while others are completely contraindicated (e.g. tetracyclines) and others are to be avoided in certain trimesters (e.g. sulphas are contraindicated in the third trimester). The choice of an appropriate antimicrobial agent must be weighed against the potential adverse outcome of a particular infection, the drug safety and spectrum of activity. PMID- 7495502 TI - Calcium antagonists. Drug interactions of clinical significance. AB - The interaction of calcium antagonists, including the dihydropyridine calcium antagonists (e.g. nifedipine), verapamil and diltiazem, with drugs from other classes has major clinical ramifications as the use of drug combinations increases in frequency. Combinations are used in the treatment of disorders ranging from hypertension to cardiac rhythm disturbances, angina pectoris and peripheral vasospastic disease. In this era of organ transplantation, drugs like cyclosporin are coming into potential conflict with an ever-growing list of drugs. Drug combinations used as part of long term therapies are also making their appearance in toxic drug reactions, including antituberculous and anticonvulsant agents. Bronchodilators and H2-blockers also fall into this category of potential culprits of combined drug toxicity, and the interactions of calcium antagonists with beta-blockers and antiarrhythmic agents are also becoming a matter of concern. PMID- 7495507 TI - Immune responses in HIV-exposed seronegatives: have they repelled the virus? AB - Over the past two years, a number of reports have described specific cellular immune responses to HIV in some individuals who, despite exposure to the virus, remain uninfected. A recent description of virus disappearance in a child born to an infected mother should be seen against the background of several previous studies that have hinted at the possibility of clearance of HIV. These findings suggest that immune-mediated clearance of virus may occur in a few fortunate individuals. PMID- 7495508 TI - The immunology of primary HIV infection: which immune responses control HIV replication? AB - In the majority of patients, HIV-1 replication is quickly and efficiently controlled after the initial burst of viremia. Recent studies have begun to elucidate the underlying immune responses that may be responsible for this dramatic reduction in viral load. These responses include those of HIV-specific CD8+ cytotoxic T lymphocytes and antibodies capable of binding the virus. PMID- 7495509 TI - Are HIV-specific CTL responses salutary or pathogenic? AB - Recently, HIV has been shown to be highly variable in patients; it is capable of escaping, or even turning off, cytotoxic T-cell responses by mutating T-cell epitopes. New antiviral drugs have revealed the enormous turnover of C4+ T cells in infected patients, but have also shown how efficiently HIV rapidly escapes such treatments. Although HIV is usually considered to be cytopathic, this is not really known. The proposal that AIDS pathogenesis reflects immunopathological consequences of anti-HIV protective CD8+ T cells has to be seriously considered. Such a pathogenesis is illustrated by CD8+ T cell mediated immunosuppression during acute infection of mice with the non-cytopathic lymphocytic choriomeningitis virus. PMID- 7495506 TI - T lymphocyte receptor deficiencies. AB - Signalling through the TCR is mediated by the cytoplasmic tails of the CD3 complex. Deficiencies in the expression of different CD3 components have lead to dramatic, yet dissimilar, effects on T-cell development and to selective deficits in peripheral T-cell subsets. Recent studies of human patients and animal models with CD3 deficiencies are providing insights into the redundant and unique roles of these molecules. PMID- 7495510 TI - Entry of microbes into the host: using M cells to break the mucosal barrier. AB - Enteric microbial pathogens interact with the gut epithelium to establish infection. Recently, it has become clear that many microorganisms that colonize or traverse the intestinal mucosa do so via the specialized M cells. Recent work has shown that Shigella flexneri and Salmonella typhimurium specifically target M cells to initiate infection of the host. PMID- 7495511 TI - Of microbes and macrophages: entry, survival and persistence. AB - The macrophage is capable of fulfilling the roles of both host cell and effector cell in the immune clearance of many microbial infections. This dual capacity has led to the evolution of intriguing mechanisms whereby pathogens ensure the establishment and maintenance of intracellular infections. The strategies developed range from passive avoidance mechanisms to extremely active modulation of cell function. Recent advances have been made in our appreciation of the processes of intramacrophage parasitization from initial ligand-receptor interactions, through establishment of a stable intracellular environment, to the maintenance of a persistent infection. PMID- 7495512 TI - Effects of IL-12 on immune responses to microbial infections: a key mediator in regulating disease outcome. AB - Recent studies have documented that the immunoregulatory functions of IL-12 may play a role in promoting endogenous protective responses during infections and/or contribute to pathology resulting from unregulated cytokine expression. Pathogen induction of IL-12 elicits interferon-gamma production by natural killer cells, which contributes to early defense during certain bacterial, parasitic, and viral infections. IL-12 also facilitates the development of T helper type 1 (Th1) lymphocytes required for late protection against bacteria, parasites, and fungi. During viral infections, however, there appear to be mechanisms independent of IL 12 for inducing protective T-cell responses. In contrast, negative regulation of IL-12 during acute infections can be a key event in the establishment of chronic infection and protection against harmful excessive cellular immune response. Under appropriate conditions, IL-12 has therapeutic efficacy for promoting defense against a variety of pathogens, and for use as a vaccine adjuvant to enhance beneficial Th1 over detrimental Th2 lymphocyte responses. This information extends knowledge about the regulation of immune responses to infectious agents, and provides new insights for the development of treatment and adjuvant strategies to potentiate beneficial or inhibit detrimental endogenous immune responses. PMID- 7495513 TI - Induction of Th2 responses in infectious diseases. AB - Recent work on T-helper (Th) cell subset maturation has focused on defining the cellular source of early IL-4, which promotes precursor (naive) CD4+ Th cells to differentiate into Th type 2 (Th2) cells, and also on the roles of counter regulatory cytokines, costimulatory signals, and antigen in the induction of Th2 responses. Results suggest that not all Th2 cells are equivalent in their ontogeny. PMID- 7495514 TI - Cytokine regulation of granuloma formation in schistosomiasis. AB - Granuloma induced by the schistosome egg is an inflammatory reaction that is tightly controlled by the interaction of T-helper cell type 1 (Th1) and Th2 cytokines produced locally. Studies focussing on the immunoregulation of the response have yielded numerous insights into the cytokine cascade that controls the formation of the lesions. It is clear from current data that the Th2 cytokines play a primary role in granuloma formation, whereas the Th1-associated lymphokine interferon-gamma acts as an endogenous downregulator of the response. Through recent advances, it may now be possible to design effective cytokine based vaccination strategies that will suppress the tissue pathology associated with this helminth infection. PMID- 7495516 TI - Mechanisms of resistance to fungal infections. AB - In the past year, significant advances have been made in our understanding of immune defenses to fungal infections, which may be instrumental in the development of rational approaches to immunodiagnosis and therapy of these infections. The highlights have been the result of the direct application of advances in molecular biology and basic immunology, particularly cytokine research, leading to improved definition of fungal antigens and increased understanding of the roles of functionally distinct T-cell subsets, the activity of which may be either host-protective or disease-promoting. PMID- 7495515 TI - The protective immune response to Mycobacterium tuberculosis. AB - Recent advances have characterized the protective immune response against Mycobacterium tuberculosis. These include identification of the phenotype of some protective cells, the antigens to which these cells respond and the cytokines produced in response to infection which modulate disease. Progress has also been made in inducing this response by vaccination. PMID- 7495517 TI - Viral mutations, TCR antagonism and escape from the immune response. AB - Persistent viruses use several mechanisms to evade the immune response, including the generation of mutations that affect TCR recognition. It has recently been reported that spontaneous mutations at TCR contact sites within individual viral epitopes in certain persistent human viruses can abrogate or antagonize the recognition of the corresponding wild-type epitope, and it has been suggested that such mutations may contribute to viral persistence. PMID- 7495518 TI - Hepatitis C virus-specific T lymphocyte responses. AB - Hepatitis C virus establishes a persistent infection in humans and chimpanzees despite virus-specific cell-mediated immune responses. Available data suggest that these responses do provide some control of ongoing hepatitis C virus replication and are an important factor contributing to chronic hepatitis. PMID- 7495520 TI - Immunodeficiency. PMID- 7495519 TI - Immunogenetics in the analysis of resistance to intracellular pathogens. AB - Recent studies have identified genes involved in resistance to intracellular pathogens. Such genes include the murine MHC class I gene, Ld (toxoplasmosis), HLA-BW53, HLA DRB1* 1302-DQ B10s01 and TNF2 (malaria), murine Nramp (toxoplasmosis, leishmaniasis and tuberculosis), gene(s) modulating the T-helper type 1 and type 2 dichotomy (leishmaniasis, leprosy and HIV infection) and the natural killer cell complex (cytomegalovirus infection). There also have been other advances in immunogenetics that have led to a better understanding of resistance to intracellular pathogens. These include effector mechanisms of immune response genes and factors modulating genetic susceptibility. Identification of genes that determine resistance/susceptibility (and their effector mechanisms) has impacted on vaccine development. Immunogenetics has been important in characterizing roles of TCR genes, superantigens, and host genes that play a role in molecular mimicry in disease pathogenesis. In addition, recent work with gene knockout, recombinant inbred or congenic, mutant, consomic, and transgenic mice, positional cloning, mouse/human gene homologies to identify candidate human resistance genes, and the rapid expansion of the gene transcription maps of the human genome, have been important in analysis of resistance to intracellular pathogens. PMID- 7495521 TI - Immunity to infection. PMID- 7495522 TI - Neuro-oncology nurses: undaunted, hopeful, and enthusiastic. PMID- 7495523 TI - Description of posterior fossa syndrome in children after posterior fossa brain tumor surgery. AB - Brain tumors are the second most common malignancy in children less than 15 years of age and the most common solid tumor of childhood. Approximately 60% to 70% of pediatric brain tumors originate in the posterior fossa. Since 1989, the two hospitals that comprise the setting for this study have treated 121 children with posterior fossa brain tumors. A postoperative syndrome, labeled posterior fossa syndrome, has been identified in certain children. This syndrome involves a variety of signs and symptoms including mutism or speech disturbances, dysphagia, decreased motor movement, cranial nerve palsies and, emotional lability. These signs and symptoms develop from an average range of 24 to 107 hours after surgery and may take weeks to months to resolve. The exact etiology of the syndrome is unknown. The purpose of this retrospective medical records review of 19 children diagnosed with posterior fossa syndrome is to describe the symptoms of the syndrome. Early recognition of this syndrome could facilitate preventive and restorative patient care, prevent subsequent complications, decrease length of hospital stays, and promote patient and family understanding of and coping with the syndrome. PMID- 7495524 TI - Pediatric brain tumors: diagnosis and management. AB - Pediatric brain tumors are the most common solid tumor and the second most common neoplasm in childhood. Confirmation of diagnosis is made by visualization of the tumor on a computed tomographic or magnetic resonance imaging scan. Treatment includes surgery followed by observation or chemotherapy and/or radiation therapy, depending on the amount of tumor removed and its histological characteristics. Recent advances in neuroradiology, neurosurgical techniques, and neuro-oncology have begun to impact the length and quality of survival of children diagnosed with brain tumors. A multidisciplinary approach in the treatment of these children is necessary with the nurses and/or nurse practitioners as the consistent members of the team often coordinating the care. PMID- 7495526 TI - The role of stereotactic radiation therapy in the management of children with brain tumors. AB - Conventional radiation therapy plays an important role in the management of intracranial tumors in children. For certain tumors radiation therapy serves as the primary mode of treatment, and for others it plays an adjuvant role with surgery and/or chemotherapy. Improvements in long-term survival rates have focused attention on the long-term sequelae of brain tumors and their treatment, and the sequelae, in turn, have become important targets for clinical investigation. Long-term side effects of particular concern in children include cranial nerve damage, memory and intellectual deficits, pituitary-hypothalamic dysfunction, demyelinization of brain tissue, and secondary malignancies. A new form of radiation therapy, stereotactic radiotherapy (SRT), merges the technologies of stereotactic surgery and conventional fractionated radiotherapy. The intent is to deliver maximum tumoricidal doses to the target while limiting the dose to normal surrounding brain tissue. The key feature of SRT is a noninvasive, relocatable immobilization system to assure accurate and reproducible positioning during planning and treatment. The headframes used for children have been modified to address their specific needs. The complexities of this process require careful preparation of patients and their families and the participation of many disciplines. Long-term follow-up will be essential to evaluate the effectiveness of this innovative treatment. PMID- 7495525 TI - Central nervous system toxicity of cancer therapy in children. AB - Cancer treatment of the central nervous system has contributed positively to long term disease-free survival from childhood cancers such as acute leukemia and medulloblastoma. However, intrathecal chemotherapy and whole brain radiation are associated with acute, subacute, and delayed toxicities. The purpose of this review article is to summarize what is known about critical events in prenatal and postnatal brain development; the pathophysiology of radiation and chemotherapy neurotoxicity and clinical manifestations and risk factors for acute, subacute, and delayed toxicities. The importance of long-term follow-up and for multidisciplinary interventions that focus on prevention as well as management of identified problems are addressed. PMID- 7495528 TI - Pharmacologic supportive care of children with central nervous system tumors. PMID- 7495527 TI - Hydrocephalus and shunts in children with brain tumors. AB - Children with brain tumors are at risk for developing hydrocephalus necessitating placement of a permanent shunt catheter. As comprehensive, interdisciplinary, multispecialty care is required for these patients, the pediatric oncology nurse must become knowledgeable about caring for children with a shunt. This article focuses on the care of the pediatric neuro-oncology patient requiring shunting for hydrocephalus, including a review of cerebrospinal fluid flow and hydrocephalus, determinants of shunt placement, complications of ventriculoperitoneal shunts, and nursing considerations. PMID- 7495530 TI - Anthranilate synthase in microorganisms and plants. AB - The enzymology of anthranilate synthase (EC 5.4.99.6) in microorganisms and plants is reviewed. Aminoacid sequences of the enzyme subunits in different species are compared, and the mechanism of reaction is discussed. PMID- 7495529 TI - Seven weeks. PMID- 7495531 TI - Tropane alkaloids and toxicity of Convolvulus arvensis. AB - Horses in a few, localized northern Colorado pastures exhibited weight loss and colic. At post mortem, intestinal fibrosis and vascular sclerosis of the small intestine was identified. The pastures where the affected horses grazed were overrun by field bindweed (Convolvulus arvensis). Bindweed from the pasture was found to contain the tropane alkaloids tropine, pseudotropine, and tropinone and the pyrrolidine alkaloids cuscohygrine and hygrine. Laboratory mice readily ate C. arvensis and exhibited a variety of abnormal clinical signs depending on the amount eaten. Similar alkaloids have been found in other Convolvulus species and cuscohygrine and calystegines (polyhydroxytropanes) have been previously reported from C. arvensis roots. This is the first report of simple tropane alkaloids in C. arvensis, a world wide problem weed. Pseudotropine, the major alkaloid, is known to affect motility and might represent a causative agent for the observed cases of equine intestinal fibrosis. PMID- 7495532 TI - Constituents and cytotoxic principles of Nothapodytes foetida. AB - A new naturally occurring alkaloid, acetylcamptothecin, together with 17 known compounds, (+)-1-hydroxypinoresinol, omega-hydroxypropioguaiacone, p hydroxybenzaldehyde, scopoletin, uracil, thymine, sitosterol, sitosteryl-beta-D glucoside, 3 beta-hydroxy-stigmast-5-en-7-one, stigmast-5-en-3 beta,7 alpha-diol, 6 beta-hydroxystigmast-4-en-3-one, sitost-4-en-3-one, linoleic acid, trigonelline, camptothecin, 9-O-methoxycamptothecin and pumiloside were isolated and characterized from the stem of Nothapodytes foetida. Among them, scopoletin, camptothecin, 9-O-methoxycamptothecin and O-acetylcamptothecin showed significant cytotoxic activity. PMID- 7495533 TI - Pregnane glycosides from Stapelia variegata. AB - Eleven new pregnane ester glycosides have been isolated from the aerial parts of Stapelia variegata. Eight of the recognized compounds were established to possess the same trioside moiety, viz. 3-O-[3-O-methyl-6-deoxy-beta-D-allopyranosyl-(1-4) beta-D- cymaropyranosyl-(1-4)-beta-D-cymaropyranoside]. These compounds were identified as: stavaroside A: 12-O-beta-angeloyl-20-O-benzoyl sarcostin; stavaroside B: 12-O-beta-angeloyl-20-O-tigloyl sarcostin; stavaroside C: 11 alpha acetoxy 2 beta-benzoxy-3 beta,8 beta, 14 beta-trihydroxy-pregn-5-ene-20-one; stavaroside D: 11 alpha-acetoxy- 12 beta-tigloxy-3 beta,8 beta,14 beta-trihydroxy pregn-5-ene-20-one; stavaroside E: 12-O-beta-benzoyl sarcostin; stavaroside F: 11 alpha-acetoxy-12 beta-acetoxy-3 beta,8 beta,14 beta-trihydroxy-pregn-5- ene-20 one; stavaroside G: 12-O-beta,20-O-diacetyl sarcostin and stavaroside H: 3 beta, 8 beta, 11 alpha, 12 beta, 14 beta-pentahydroxy-pregn-5- ene-20-one. The other three compounds were shown to possess the same tetraside sugar moiety, viz. 3-O [beta-D-glucopyranosyl- (1-4)-3-O-methyl-6-deoxy-beta-D-allopyranosyl-(1-4)-beta D-cymaropyra nosyl- (1-4)-beta-D-cymaropyranoside]. These compounds were identified as: stavaroside I: 1 alpha, 12 beta-angeloxy and benzoxy-3 beta,8 beta,14 beta-trihydroxy- pregn-5-ene-20-one; stavaroside J: 11 alpha-acetoxy-12 beta-benzoxy-3 beta, 8 beta,14 beta-trihydroxy-pregn-5-ene-20-one and stavaroside K: 11 alpha-acetoxy-12 beta-tigloxy-3 beta,8 beta,14 beta-trihydroxy-pregn-5-ene- 20-one. The structural elucidation of the isolated compounds was aided significantly on the basis of the chemical and spectral evidence. The decisive assignments of the ester positions were based on the Inverse Detected Heteronuclear Multiple Bond Connectivity (HMBC) experiments. PMID- 7495534 TI - Anacardoside from the seeds of Semecarpus anacardium. AB - From the seeds of Semecarpus anacardium, a new phenolic glucoside, anacardoside, was isolated, and its structure and configuration were elucidated by a combination of NMR techniques as 1-O-beta-D-glucopyranosyl-(1-->6)-beta-D glucopyranosyloxy-3-hydro xy-5-methylbenzene. PMID- 7495535 TI - Method for constructing internal standards for use in competitive PCR. PMID- 7495536 TI - Isolation of plasmid DNA from mycobacteria using a resin-based alkaline lysis kit. PMID- 7495537 TI - Lower incubation temperature increases yield of insect genomic DNA isolated by the CTAB method. PMID- 7495538 TI - Background reduction in northern analysis by preabsorption of digoxigenin-labeled riboprobes. PMID- 7495539 TI - RNA extraction from gastrointestinal tract and pancreas by a modified Chomczynski and Sacchi method. PMID- 7495540 TI - New technique for precise alignment of an RNA differential display gel with its film image. PMID- 7495542 TI - Plasmid ligation using a sacrificial insert. PMID- 7495541 TI - Protein immunoblotting of stained gels. PMID- 7495543 TI - E. coli beta-glucuronidase (GUS) as a marker for recombinant vaccinia viruses. PMID- 7495544 TI - Preparation of recombinant baculovirus by transfection of a ligated cDNA fragment without prior plasmid amplification in E. coli. PMID- 7495545 TI - Reporter enzyme-nitrilotriacetic acid-nickel conjugates: reagents for detecting histidine-tagged proteins. PMID- 7495546 TI - Dual temperature in situ hybridization. PMID- 7495547 TI - Improvement of fluorescence in situ hybridization (RNA-FISH) on human paraffin sections by propidium iodide counterstaining. PMID- 7495548 TI - Mini-preparation of total RNA for RT-PCR from cultured human cells. PMID- 7495549 TI - Simple protocol for rapid, direct sequencing of immunoglobulin variable-region genes. PMID- 7495550 TI - Electrotransfer of Long Ranger sequencing gels using a methanol-TBE buffer. PMID- 7495551 TI - Preparation of high molecular weight genomic DNA from nuclei of woody plants. AB - We have developed a rapid and reliable method for preparation of high molecular weight genomic DNA from sweet orange (Citrus sinensis) suitable for subsequent digestion by rarely cutting restriction enzymes and then separation by pulsed field gel electrophoresis (PFGE). Methods previously described for preparation of plant DNA prior to PFGE involved protoplast isolation, a procedure that can be inefficient and time-consuming for several plant species. Nuclei isolated from plant tissues were embedded into agarose blocks and treated to release DNA, which was cleaved by restriction enzymes and then submitted to PFGE. One gram of fresh leaves gave approximately 15 micrograms of high molecular weight genomic DNA (> 2000 kbp). Within-gel hybridizations were used instead of classical Southern blotting, and the resulting signals were adequate when they were compared with those obtained with DNA prepared from crude ground leaf tissues. PMID- 7495552 TI - Single-step protocol for preparation of plant tissue for analysis by PCR. AB - PCR has many applications in the isolation and analysis of plant DNA. The influence of salt and EDTA concentration, pH, incubation time and temperature on the preparation of plant material for PCR was evaluated. A general single-step method was developed in which a small amount of plant tissue was heated in a simple solution. The DNA in the supernatant was found to be suitable for most PCR applications including arbitrarily primed PCR (random-amplified polymorphic DNA) and PCR with specific primers for both single- and multiple-copy genes. The technique is much simpler than those generally used for plant DNA preparation and was successful with tissues from a wide range of species. PMID- 7495553 TI - Primer-induced labeling of pea and field bean chromosomes in situ and in suspension. AB - A protocol for primed in situ DNA labeling (PRINS) was optimized for pea (Pisum sativum L.) and field bean (Vicia faba L.) chromosomes attached to coverslips. Cloned DNA or synthetic oligonucleotides were used as probes for repetitive DNA sequences (rDNA, Fok-element) and different reaction conditions were tested to achieve the highest specific signal-to-background ratio. A procedure based on direct labeling by fluorescein-dUTP was compared with an indirect one using digoxigenin detected by fluorescently labeled antibody. Under optimal conditions, strong and specific signals were obtained exclusively on chromosome regions known to contain respective DNA sequences. Compared to the direct labeling, significantly stronger signals were obtained when the indirect procedure was used. Both types of labeling were successfully applied to chromosomes in suspension and were shown to produce signals comparable to that obtained with chromosomes attached to coverslips. It is expected that primed in situ DNA labeling en suspension (PRINSES) will provide a basis for flow-cytometric discrimination and sorting of otherwise indistinguishable chromosomes according to their specific fluorescent labeling. PMID- 7495554 TI - High-resolution SSCP analysis using polyacrylamide agarose composite gel and a background-free silver staining method. AB - We describe here an improved methodology for single-strand conformation polymorphism (SSCP) analysis of PCR products. The method utilizes a polyacrylamide agarose composite gel and background-free silver staining. In comparison with conventional pure polyacrylamide gel for PCR-SSCP analysis, the composite gels have a much greater mechanical strength and improved resolution. The background staining commonly seen in many silver staining protocols has been eliminated by incorporation of thiosulfate, which can prevent nonspecific deposits of silver salts. As shown in our titration tests, the composite gel and background-free silver staining together have allowed clear identification of point mutations in samples containing as little as 5% of the target sequences, the same sensitivity achieved by radioactive labeling methods. PMID- 7495555 TI - Use of vaccinia virus poly(A) polymerase for RNA 3'-end labeling with a chain terminating nucleotide or a short 3' homopolymer tract. AB - Conditions are described for the 3'-end labeling of RNA with 32P 3'-dATP (3' deoxyadenosine-5'-triphosphate), a chain-terminating nucleotide, using the poly(A) polymerase (PAP) encoded by vaccinia virus. Reaction time, divalent cation species and concentration, and the requirement for both subunits of the PAP were investigated. In the presence of Mn2+, vaccinia PAP is able to tail RNA primers with tracts of 3'-oligo(U), oligo(C) and oligo(G). Conditions for the addition of labeled 3'-homopolymer tracts were characterized. The use of low nucleotide concentrations in this study revealed an apparently fixed divalent cation concentration optimum of 0.1 mM, distinct from the previously noted requirement for a 1:1 divalent cation:NTP complex. This indicates a possible requirement for multiple divalent cations in nucleotidyl transfer by vaccinia PAP. PMID- 7495556 TI - Automated DNA purification and amplification from blood-stained cards using a robotic workstation. AB - A new method for the purification of DNA on blood-stained cards was developed. This method was implemented into a high-throughput automated system using a Biomek 1000 robotic workstation. In addition, the processes of DNA purification and amplification were coupled into a completely automated and uninterrupted prototype system, and the resultant PCR products generated by this system were subjected to automated desalting for capillary electrophoresis analysis. PMID- 7495557 TI - In situ assessment of beta-hexosaminidase activity. AB - We have adapted two methods to evaluate the beta-hexosaminidase (HEX) enzymatic activity in cultured cells, based on the use of (i) the fluorogenic substrate 4 methylumbelliferyl-6-sulfo-2-acetamido-2- deoxy-beta-D-glucopyranoside (MU-GlcNAc 6-SO4) and (ii) the naphthol AS-BI-N-acetyl-beta-D-glucosaminide and hexazotized pararosaniline. We demonstrate that both methods could be used for the HEX isoenzymes by comparing wild-type and mutant human fibroblast cell lines, deficient for either an alpha or beta subunit from Tay-Sachs and Sandhoff patients. This in situ cytochemical assessment of HEX activity offers a rapid evaluation to study the expression of this enzyme in a heterogeneous cell population such as in gene transfer experiments. PMID- 7495558 TI - Using oligonucleotide probe arrays to access genetic diversity. AB - As the Human Genome Project and related efforts identify and determine the DNA sequences of human genes, it is important that highly reliable and efficient mechanisms are found to access individual genetic variation. It is only through a greater understanding of genetic diversity that the true benefit of the Human Genome Project will be realized. One approach, hybridization to high-density arrays of oligonucleotides, is a fast and effective means of accessing this genetic variation. Light-directed chemical synthesis has been used to generate miniaturized, high-density arrays of oligonucleotide probes. Application-specific oligonucleotide probe array designs have been developed for the rapid screening of characterized genes. Dedicated instrumentation and software have been developed for array hybridization, fluorescence detection and data acquisition and analysis. In a specific and challenging application, oligonucleotide probe arrays have been used to screen the reverse transcriptase and protease genes of the highly polymorphic HIV-1 genome to explore genetic diversity and detect mutations conferring resistance to antiviral drugs. Results from this application strongly suggest that oligonucleotide probe arrays will be a powerful tool for rapid investigations in sequence checking, pathogen detection, expression monitoring and DNA molecular recognition. PMID- 7495559 TI - Accuracy of automated DNA sequencing: a multi-laboratory comparison of sequencing results. AB - A double-stranded (ds)DNA template of "unknown" sequence was distributed to approximately 80 core DNA sequencing laboratories by the Association of Biomolecular Resource Facilities (ABRF) for automated DNA sequence analysis. Forty-four different facilities responded with 83 usable sequence submissions. These sequences were grouped by both sequencing protocol (dye-primer or dye terminator) and whether manually edited or not. The sequences were aligned with the known sequence, and the number of correct base calls, insertions, deletions, no-calls and miscalls were determined for each group. The dye-primer sequencing protocol provided the longest and most accurate sequence. The edited dye-primer data were > 95% accurate out to 400-450 bp, while the edited dye-terminator data could call only 300-350 bases at this accuracy. However, 75% of the laboratories in this sampling preferred the dye-terminator protocol, presumably because of its versatility and convenience. Laboratories that manually edited the automatically called data were able to obtain an additional 100 bases of good sequence when the dye-primer protocol was used. Surprisingly though, editing of dye-terminator results did not increase the amount of good sequence, although the dye-terminator protocol had a superior base-calling ability within the first 100 bases of called sequence. PMID- 7495561 TI - Optimization of Applied Biosystems 373 sequencer stretch modification. AB - We have optimized the conditions for using the Stretch modification for the Applied Biosystems 373 Automated DNA Sequencers for sequencing double-stranded DNA using 34-cm well-to-read and 48-cm well-to-read configurations. With the manufacturer's recommended settings, uneven spacing within the first 100 bases was observed, which led to miscalls, insertions and deletions in the analyzed data. A significant decrease in accuracy for reads greater than 400 bases was also observed. Various gel concentrations were tested to improve the base spacing for the first 100 bases while maintaining accuracy and usable length of data. A longer average usable length and better resolution of smaller fragments were achieved by increased acrylamide concentration coupled with increased wattage. Using the Applied Biosystems CATALYST 800 Molecular Biology LabStation, Taq dye primer cycle sequencing reactions were optimized for -21 M13 and M13RP1 primers to produce a more even distribution of dye-labeled fragments that increased the overall signal strengths and decreased background signal. These reaction products, run on the Stretch sequencers using the new gel conditions, provided longer reads with increased reliability and accuracy of the data. PMID- 7495562 TI - Sequence-specific purification of nucleic acids by PNA-controlled hybrid selection. AB - Using an oligohistidine peptide nucleic acids (oligohistidine-PNA) chimera, we have developed a rapid hybrid selection method that allows efficient, sequence specific purification of a target nucleic acid. The method exploits two fundamental features of PNA. First, that PNA binds with high affinity and specificity to its complementary nucleic acid. Second, that amino acids are easily attached to the PNA oligomer during synthesis. We show that a (His)6-PNA chimera exhibits strong binding to chelated Ni2+ ions without compromising its native PNA hybridization properties. We further show that these characteristics allow the (His)6-PNA/DNA complex to be purified by the well-established method of metal ion affinity chromatography using a Ni(2+)-NTA (nitrilotriactic acid) resin. Specificity and efficiency are the touchstones of any nucleic acid purification scheme. We show that the specificity of the (His)6-PNA selection approach is such that oligonucleotides differing by only a single nucleotide can be selectively purified. We also show that large RNAs (2224 nucleotides) can be captured with high efficiency by using multiple (His)6-PNA probes. PNA can hybridize to nucleic acids in low-salt concentrations that destabilize native nucleic acid structures. We demonstrate that this property of PNA can be utilized to purify an oligonucleotide in which the target sequence forms part of an intramolecular stem/loop structure. PMID- 7495560 TI - Allele-specific HLA-DRB1 amplification of forensic evidence samples with mixed genotypes. AB - A major problem in analyzing forensic casework samples is the presence of genetic material from more than one individual in the material evidence. For instance, in sexual assault cases the evidence (vaginal swabs) usually contains a majority of vaginal epithelial cells and varying amounts of sperm cells from the perpetrator. Samples with mixed genotypes are also common among other biological evidence materials such as nail scrapes and mixed bloodstains. We have developed an allele specific amplification system for the highly polymorphic HLA class II DRB1 locus that permits the detection of individual alleles in a sample with mixed genotypes, independent of the initial frequency of the alleles. Using a set of eight allele-specific amplification primers and typing the amplified fragments with sequence-specific probes, most of the 60 DRB1 alleles can be resolved. The method is highly specific and sensitive, with the potential for amplifying 15 copies of a particular allele in a background of 3 x 10(5) copies of other alleles. The method was successfully applied to three forensic cases, where the material evidence consisted of sperm stains on panties, nail scrapes and bloodstains on skin. Thus the DRB1 allele-specific amplification system can be employed for the unambiguous determination of the presence of individual alleles in materials suspected to contain mixed genotypes, even when the alleles of interest constitute only a small fraction of the total DNA. PMID- 7495564 TI - Quantitative nonradioactive CAT assays using fluorescent BODIPY 1 deoxychloramphenicol substrates. AB - We report the development of fluorescent BODIPY 1-deoxychloramphenicol substrates for chloramphenicol acetyltransferase (CAT). These substrates not only simplify and improve quantitation of CAT activity but also extend the linear range of detection. Because the 1-deoxychloramphenicol derivatives have only one acetylation site, the enzyme reaction creates only one product, whereas chloramphenicol and its fluorescent derivatives produce three acetylated products, each of which accumulates at a different rate. Thus, 1 deoxychloramphenicol substrates eliminate the need to account for multiple products and provide a method in which product formation corresponds directly to enzyme activity. These nonradioactive substrates also allow researchers to streamline the standard thin-layer chromatography separation procedure: visible results can be obtained within minutes and quantitative results in a few hours. The sensitivity of CAT assays using fluorescent 1-deoxychloramphenicol substrates is comparable to that achieved using [14C]chloramphenicol--between 10(-5) and 10( 6) units of activity in a 1-h reaction--and the linear range extends through 3.6 or more orders of magnitude. We expect that CAT assays employing BODIPY 1 deoxychloramphenicol CAT substrates will be a valuable improvement over other methods currently in use. PMID- 7495563 TI - Sequencing 500 kb of yeast DNA using a GATC 1500 Direct Blotting Electrophoresis System. AB - We have developed efficient protocols for the construction of shotgun libraries and for high-throughput DNA sequencing at low cost using the optimized GATC 1500 Direct Blotting Electrophoresis system with colorimetric detection on nylon membranes. We can now sequence entire cosmids without any primer walking at low coverage (8-fold to 9-fold). We have demonstrated the efficiency of our protocols while sequencing 500 kb of yeast DNA with the GATC 1500 system. The methods described here have allowed us to achieve the highest data rate within the framework of the European Communities yeast sequencing program. In 1993, only two people were involved in generating and submitting 156,274 bp of final sequence data. Thereof, 39,360 bp have been verified independently in other laboratories, and no sequencing error has been found. PMID- 7495565 TI - Cell volume regulated transporters of compatible osmolytes. AB - Virtually all cells respond to hypertonicity by accumulating certain small organic solutes (compatible osmolytes) that, in contrast to intracellular ions, do not perturb macromolecular function. Several important compatible osmolytes are accumulated by coupled transport. Transcription of genes encoding these cotransporters is increased by hypertonicity and a tonicity-responsive enhancer element has been identified. When cells return to an iso-osmotic environment, osmolytes are rapidly lost through a pathway that current evidence indicates may be a volume-sensitive chloride channel. PMID- 7495566 TI - Aquaporin water channels: unanswered questions and unresolved controversies. AB - The long-standing biophysical question of how water crosses plasma membranes has been answered by the recent discovery of the aquaporins. Identification of this large family of membrane water-transport proteins has generated new questions about the physiological functions, tissue distributions, and regulatory mechanisms of individual aquaporins. The fast pace of developments in this field has also resulted in major discrepancies in published reports which warrant resolution. PMID- 7495567 TI - The extracellular calcium receptor. AB - The importance of intracellular calcium in regulating cell function is well recognized. No less important, but less well understood (and probably appreciated), is the fundamental role played by extracellular calcium, Ca2+o, in the modulation of cell function. The recent cloning of Ca2+o-sensing, G-protein coupled receptors from bovine (and human) parathyroid and rat kidney (and brain) has clearly demonstrated that Ca2+o can function as a traditional 'first messenger'. The identification of 'inactivating' and 'activating' mutations in this Ca2+o-sensing receptor in two hypercalcemic disorders and in an autosomal dominant form of hypocalcemia, respectively, has underscored the physiological relevance of this receptor in Ca2+ homeostasis in man. These advances have significantly enhanced our understanding of the molecular mechanisms involved in extracellular calcium sensing in parathyroid and kidney. Moreover, the localization of the Ca2+o-sensing receptor in tissues previously not known to have Ca2+o-sensing capability has suggested novel and potentially quite important roles for Ca2+o in regulating the function of cells not apparently directly involved in Ca2+ homeostasis. PMID- 7495568 TI - Molecular characterization of the epithelial Na-K-Cl cotransporter isoforms. AB - Recent advances in the molecular characterization of specific isoforms of the Na K-Cl cotransporter have allowed rapid progress in the study of the structure, function, and regulation of these members of a family of Cl-dependent cation cotransporters. Two distinct isoforms have been identified, one from Cl(-) secretory epithelia and another found specifically in the diluting segment of the vertebrate kidney, a Cl(-)-absorptive epithelium. The discovery of three alternatively spliced variants of the absorptive isoform, which differ only by 31 amino acids and which appear to be differentially distributed within the mammalian thick ascending limb of the loop of Henle, highlight this spliced region as an important functional component of the protein. PMID- 7495569 TI - Chloride channels of intracellular organelles. AB - Chloride channels are present in a variety of intracellular organelles (Golgi, endosomes, endoplasmic reticulum, and sarcoplasmic reticulum) where they serve largely to shunt the membrane potential created by other ion-translocating processes. Electrophysiological studies have shown that the Cl- channels of the endoplasmic and sarcoplasmic reticula facilitate the efflux of Ca2+. In the Golgi and some endosomes, the open Cl- channels (probably the cystic fibrosis transmembrane conductance regulator) favor accumulation of H+. PMID- 7495570 TI - Structure and function of fusion pores in exocytosis and ectoplasmic membrane fusion. AB - Several proteins involved in exocytosis have been identified recently, but it is still completely unclear which molecules perform the fusion event itself. Although in viral fusion the fusion proteins are known, even there the molecular mechanism remains controversial. Investigation of single fusion events by electrophysiological techniques together with fluorimetric measurements have now provided some insight into the properties of the first aqueous connection, the fusion pore. This pore has an initial size similar to an ion channel and allows movement of lipids only after it has substantially expanded, indicating that it is initially not a purely lipidic structure, but incorporates lipids when it expands. Although neurotransmitter release may occur through narrow transient fusion pores, the fusion pore of synaptic vesicles probably expands vey rapidly, making it unlikely that secretion is performed by rapid exo/endocytosis without full fusion under normal conditions. Recent recordings from small membrane patches have made it possible to resolve fusion events from vesicles as small as synaptic vesicles. Future experiments using excised patches may provide an approach to identify the molecular machinery of exocytotic membrane fusion. PMID- 7495571 TI - Membranes and sorting. PMID- 7495572 TI - Quality control in the secretory pathway. AB - Whereas newly synthesized proteins that have acquired a properly folded and assembled structure are transported from the endoplasmic reticulum to their final destinations, incompletely folded and assembled proteins are, as a rule, retained and eventually degraded. The molecular mechanisms of this unique molecular sorting phenomenon, called 'quality control', have been illuminated by recent studies. PMID- 7495573 TI - Sorting and retrieval between the endoplasmic reticulum and Golgi apparatus. AB - The highlight of the past year was the demonstration that retrieval of endoplasmic reticulum membrane proteins containing the di-lysine motif involves COPI coat proteins. Other findings contributed to the debate about the nature of the 'intermediate compartment' between the endoplasmic reticulum and the Golgi apparatus, and the mechanism by which transported proteins are concentrated at this step. PMID- 7495574 TI - The role of coat proteins in the biosynthesis of secretory proteins. AB - The biosynthesis of secretory proteins requires vesicle-mediated transport between the organelles of the secretory pathway. Biochemical and genetic analysis of the secretory pathway has identified two non-clathrin coats--COPI and COPII- that drive the formation of vesicles that mediate transport between the endoplasmic reticulum and the Golgi apparatus, and through the compartments of the Golgi. Recently, a molecular description of the subunits of these coats and the development of biochemical reagents to study their function has yielded new information on how these proteins share the task of organizing vesicle traffic early in the secretory pathway. PMID- 7495575 TI - Protein transport to the yeast vacuole. AB - Genetic and biochemical analyses of yeast vacuolar protein localization have identified more than 40 gene products that play a role in this process. Included among these components are a sorting receptor, a protein kinase, a phosphatidylinositol kinase, small GTP-binding proteins and a dynamin-like GTPase. Some of these gene products are homologous to proteins required for sorting and transport at other stages of the secretory and endocytic pathways. Others appear to be required for unique functions in the vacuolar protein localization pathway. Recent studies have helped to define the role that each of these components plays in vacuolar protein localization and have offered new insights into the molecular mechanisms of protein sorting. PMID- 7495576 TI - Membrane transport in the endocytic pathway. AB - Despite controversies and debates, some fundamental properties of endosomes become apparent when comparing results from in vivo and in vitro strategies used to study endosomal membrane traffic. In addition, recent studies are starting to unravel the complex organization of early endosomes, in particular along the route followed by recycling receptors. PMID- 7495577 TI - Lonely MHC molecules seeking immunogenic peptides for meaningful relationships. AB - The association between antigenic peptides and MHC class II molecules represents a critical event in the initiation of the immune response to extracellular antigens. Understanding the molecular basis of antigen processing requires the characterization of the intracellular compartments, or 'singles bars', in which immunogenic peptides are generated and loaded onto class II molecules. In the past year, something of a breakthrough occurred with the identification of specialized compartments that host antigen processing and/or peptide loading, designated 'MHC class II compartment' and 'class II vesicles'. It is becoming increasingly clear, however, that these compartments are themselves heterogeneous and not always distinct from conventional endosomes and lysosomes. PMID- 7495579 TI - SNAREs and the specificity of transport vesicle targeting. AB - The fidelity of membrane trafficking is mediated by specific interactions between transport vesicles and their target membranes. SNAREs (SNAP receptors) are cytoplasmically oriented membrane proteins that form complexes implicated in transport vesicle targeting. Advances in the past year have provided insight into the fundamental mechanisms underlying the formation, regulation, and function of SNARE complexes. PMID- 7495578 TI - The emergence of clathrin-independent pinocytic pathways. AB - Receptor-mediated endocytosis via clathrin-coated vesicles is by far the best characterized example of pinocytosis. It has been suggested that clathrin-coated vesicles mediate all pinocytosis in mammalian cells. This is still a matter of debate, however, and recent results provide strong evidence for 'clathrin independent' pinocytic pathways. The selective regulation of these alternate endocytic pathways and the identification of receptors targeted to them provide new tools for the functional and mechanistic characterization of clathrin independent pinocytosis. PMID- 7495580 TI - Molecular motors, membrane movements and physiology: emerging roles for myosins. AB - Myosins are a large family of structurally diverse mechanoenzymes which, upon interaction with actin filaments, convert energy from ATP hydrolysis into mechanical force. Consistent with the ubiquitous association of actin with membranes, many of these novel myosins are membrane-associated. In the past two years, evidence has emerged that suggests roles for actin-based molecular motors in a wide range of membrane phenomena such as cell locomotion, phagocytosis, secretion, organelle transport, signal transduction and mechanoregulation of membrane protein function. PMID- 7495581 TI - Membrane permeability. PMID- 7495582 TI - Membranes and sorting. PMID- 7495586 TI - Survival and quality of life in patients with protracted recovery from cardiac surgery. Can we predict poor outcome? AB - Of all the 2256 adult cardiac surgical patients operated upon during a 12-month period from 1st February 1992 in three units, only 162 (7.2%) spent more than 48 h in the intensive care unit (ICU) (median 6 days, range 3-90). There were 47 deaths in ICU, 7 more before hospital discharge, and a further 10 before the study end-point of one year after surgery. All 98 1-year survivors were at home with 86 of them reporting their quality of life, on formal evaluation, to be within the reference range which we have established for a less complicated cohort of cardiac surgical patients. Prospectively collected physiological measurements were used in a mathematical model to test how well we could predict which patients will die and which of the survivors have a poor quality of life. The algorithm performs well for cardiac surgery patients with a specificity of 98%. If treatment had been withdrawn when death or poor quality of life became predictable, the maximum number of ICU bed days that could be freed was of the order of 2%. The plight of these patients is distressing, but most survive and do well and they are infrequent compared with the large majority who survive to leave hospital after a short ICU stay. PMID- 7495584 TI - Reduction of reoxygenation injury and nitric oxide production in the cyanotic immature heart by controlling pO2. AB - Reintroduction of high levels of molecular oxygen after a hypoxic period is followed by a burst of nitric oxide (NO), peroxynitrite, and oxygen free radicals, which are highly cytotoxic. This study tests the hypotheses that a) controlled reoxygenation of cyanotic immature hearts when starting cardiopulmonary bypass (CPB) with high pO2 pressure of oxygen produces a reoxygenation injury, and b) this oxygen-related damage is avoidable by controlling the circumstances of the reoxygenation period (controlled reoxygenation). Of 40 immature piglets (2-3 weeks), 5 normoxic instrumented piglets served as control, and 6 underwent 1 h of CPB including 30 min of aortic clamping with blood cardioplegic (BCP) arrest without preceding hypoxia (BCP control). Twenty-nine others were made hypoxic (arterial pO2 20-30 mmHg) for up to 2 h by lowering the forced inspiratory oxygen (FiO2) on a ventilator. They were then reoxygenated on CPB as follows, 1) abrupt reoxygenation at pO2 400 mmHg in 5, (Reox), 2) gradual increase in pO2 from 30 to 400 mmHg in 5 (Graded Reox), both without BCP arrest, 3) starting CPB at different pO2 levels (hyperoxic, normoxic or hypoxic) for 5 min, followed by BCP arrest (Reox+BCP: pO2 > 400, 100 or 20-30 mmHg), in 19 others. Reoxygenation on CPB at pO2 more than 400 mmHg depressed contractility (endsystolic elastance [Ees] to 25 +/- 5% of control; P < 0.05), accompanied by reduced antioxidant reserve capacity [AORC] (P < 0.05 vs control), which was only slightly improved by Graded Reox (Ees 34 +/- 4%, P < 0.05 vs control).(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7495585 TI - The Edinburgh Cardiac Surgery Score survival prediction in the long-stay ICU cardiac surgical patient. AB - Predictors of outcome in long-stay patients following cardiac surgery have hitherto been ill defined. The aims of this study were to test the Parsonnet pre operative scoring system and to define a scoring system for inhospital mortality applicable post-operatively to strengthen the clinical decision-making process. Following case note review of 262 consecutive patients who stayed 7 days or more in intensive care, a total of 110 pre-, intra- and post-operative factors were documented. In this long-stay group the Parsonnet score was confirmed to be predictive of 30 day mortality. Univariate analysis identified significant association between mortality in the Intensive Care Unit (ICU) and the following: inotrope days, (defined as number of inotropes times number of days) ventilation, units of platelets (P = < 0.00001), chest reopening, fresh frozen plasma units (P < 0.002), total parenteral nutrition, noradrenaline, Parsonnet score (P = 0.005), dopamine, bypass time, vasodilators, intra-aortic balloon counterpulsation, enteral nutrition and other major surgery (P < 0.05). Stepwise logistic regression on these significant factors was used to produce the Edinburgh Cardiac Surgery Score (ECS) applicable from Day 10 onwards in the intensive care unit: ECS Score = (Inotrope days) +2 (Ventilation) + (Platelets) + (Parsonnet) -3. The ECS score may be a useful predictor of ICU mortality probability for cardiac surgical patients requiring 10 days or more intensive care and is presently undergoing prospective evaluation in our centre. PMID- 7495583 TI - Continuous warm versus intermittent cold blood cardioplegia for coronary bypass surgery in patients with left ventricular dysfunction. AB - Between October 1991 and March 1994, 108 consecutive patients with moderate to severe left ventricular dysfunction underwent non-emergency isolated coronary artery surgery under the care of one surgeon (A.R.). They were prospectively randomised to receiving either intermittent cold (Group 1-50 patients) or continuous warm (Group 2-58 patients) blood cardioplegia for myocardial protection. There were no significant differences in clinical outcome between the two groups, as judged by operative mortality, rates of perioperative myocardial infarction, the serum CKMB isoenzyme level at 2 and 18 h after operation, need for circulatory support, postoperative neurological deficit, or duration of hospital stay. Group 2 patients required significantly more potassium (68 vs 29 mmol, P < 0.001) to maintain diastolic arrest and also had higher serum potassium levels after removal of the cross-clamp (P < 0.001). However, sinus rhythm returned spontaneously with greater frequency (91.2% vs 45.8%, P < 0.001) in Group 2 patients. In conclusion this report suggests that retrograde continuous warm blood cardioplegia provides comparable myocardial protection to that achieved with retrograde intermittent cold blood cardioplegia in patients with moderate to severe left ventricular dysfunction undergoing isolated coronary artery surgery. PMID- 7495587 TI - Quality of care in adult heart surgery: proposal for a self-assessment approach based on a French multicenter study. AB - A prospective study was carried out from January to April 1993 in 42 centers to assess the quality of care in adult heart surgery in France. One hundred eight parameters were collected (i.e., risk factors, surgery, postoperative course) for 7181 patients. Eight simple, objective factors were selected using a multivariate analysis. Point weighting was assigned to each factor. A simple cumulative score was thereby established to classify patients within four incremental risk groups (respective distribution of patients and mortality were 38.6%, 26.5%, 21.3%, 13.5% and 2%, 3.9%, 6.1%, 21.4%). This score was highly predictive for mortality and severe morbidity. Objectivity and independence of the observer, coupled with a reliable score, are needed for meaningful assessment of the quality of care. Administrative supervision and/or comparison of raw center results therefore remain controversial. A national data base was developed as a part of this study, that includes anonymous information from two-thirds of all cardiac surgery cases. Nation-wide results for France allow each center freely to assess its results. This self-assessment approach is, in our view, the most accurate way of quality of care assessment. A pan-European system using a similar approach should be developed. PMID- 7495588 TI - The valve choice in tricuspid valve replacement: 25 years of experience. AB - This study reviews 146 consecutive patients who underwent tricuspid valve replacement (TVR) with 69 bioprostheses (porcine and bovine pericardial) and 77 mechanical ball, disc or bileaflet valves between 1967 and 1987. The mean age was 51.4 +/- 12.1 years. Preoperatively, 97% were in New York Heart Association (NYHA) functional class III or more and over 40% had undergone previous cardiac surgery. Hospital mortality was high (16.1%). Incremental risk factors for hospital death were preoperative icterus (P < 0.01), hepatomegaly (P = 0.02), NYHA functional class IV (P = 0.02) and male sex (P = 0.04) (univariate analysis). Ninety-eight percent of the hospital survivors were followed up for a mean of 92 months. Cumulative follow-up added up to 955 patient-years. There were 70 late deaths. The actuarial survival rate was 74% at 60 months and less than 25% at 14 years. Incremental risk factors for late death indicated by univariate analysis were the type of tricuspid prosthesis (Smel-off-Cutter and Kay-Shiley prostheses) (P = 0.04), the type of operative myocardial protection (normothermia and coronary perfusion) (P = 0.05) and preoperative NYHA functional class IV (P = 0.05). We conclude that TVR carries a high operative risk and poor long-term survival, both influenced by preoperative and perioperative variables. Bearing in mind the poor prognosis for TVR, we prefer a large-sized bioprosthesis, in view of its initial good durability and low risk of valve-related events. However, in patients with good life expectancy, a bileaflet mechanical prosthesis may be an acceptable alternative. PMID- 7495589 TI - Right ventricular outflow reconstruction with homografts. AB - Ninety children and young adults underwent right ventricular outflow tract (RVOT) reconstruction with aortic or pulmonary homografts from May, 1989 to May, 1994. The patients were divided into three groups according to preoperative diagnosis: RVOT obstructions with ventriculo-arterial (VA) concordance (61), RVOT obstructions with VA discordance (18) and truncus arteriosus (11). Of the reconstructions, 52% were reoperations. A pulmonary homograft was used by preference (85% in the concordant group and 33% in the discordant group). One patient died after homograft correction (hospital mortality 1.1%). The mean follow-up was 32 +/- 22 months. One patient died after 10 months due to congestive heart failure and obstructive pulmonary hypertension. All other patients were in NYHA classes I-II. Three patients (two discordant and one truncus correction) underwent reoperation because of aortic homograft dysfunction and calcification. The incidence of significant (> or = 40 mmHg) gradient across the reconstruction found at the last echocardiographic control was 4% in the concordant, 28% in the discordant, and 18% in the truncus, group. Echocardiographic evidence of moderate (grade 2-3/4) pulmonary regurgitation (PR) was noticed in 24 and 13% of the concordant and discordant groups, respectively. From this medium-term experience it is concluded that (1) the medium-term performance of cryopreserved homografts is excellent, even in young patients, (2) corrections in patients with VA discordance need close follow-up, (3) the use of pulmonary homografts, especially in VA concordant lesions, should be encouraged, (4) longer-term follow-up is necessary to confirm the superiority of the cryopreserved homograft when compared to the porcine xenograft in the RVOT. PMID- 7495590 TI - Surgery for tetralogy of Fallot at less than six months of age. Is palliation "old-fashioned"? AB - There is still no consensus regarding the most appropriate protocol for managing symptomatic patients with tetralogy of Fallot (TOF) presenting during early infancy. From January 1987 to April 1994, 83 patients with TOF were operated on at less than 6 months of age. Sixty patients (mean age 109 +/- 5 days) underwent primary repair and 23 (mean age 72 +/- 10 days) underwent initial palliation with a modified Blalock-Taussig shunt (MBTS) in 22 cases. No strict protocol was used but patients who received initial palliation had lower weight, smaller pulmonary annulus or had an anomalous coronary artery. Nineteen of the patients initially palliated underwent repair at a median of 13 months after palliation. During this interval, the pulmonary annulus size increased from a Z-value of -3.0 +/- 0.4 at palliation to -1.6 +/- 0.7 at repair (P = 0.06) and the summed diameters of pulmonary artery branches from -2.2 +/- 0.4 to -1.6 +/- 0.7 (P = 0.2). There were no operative deaths among the repaired patients, but two patients died early after shunt insertion. Transannular patch was required in 58% of the patients regardless of the management protocol. Actuarial survival rate and freedom from reoperation at 48 months were 98.0% and 77.4% (P = 0.003) and 87.6% and 90.0% in primarily repaired and initially palliated patients, respectively. Primary repair of TOF at the time of presentation is the preferred approach regardless of age.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7495591 TI - Maximal thymectomy for myasthenia gravis. AB - Maximal thymectomy was carried out in 48 patients with myasthenia gravis (MG). There were 18 males and 30 females. Thymic hyperplasia was found in 38, and atrophic thymus in 8, patients. Two patients had thymoma. In the non-thymomatous myasthenia gravis complete remission was achieved in 16 patients (34.8%) and pharmacological remission in 20 patients (43.5%) thus giving a total remission in 36 (78.3%) patients. Six patients (13%) improved. There was no improvement in four patients. Thus, the overall benefit from thymectomy was 91.4% in this series. We found that sex, age at onset of disease and steroid therapy influenced the outcome of thymectomy. On the other hand, duration of disease, anti acetylcholine receptor (AntiAchR) antibodies and thymic histology did not have any bearing on the complete remission rate. PMID- 7495592 TI - Successful construction of a mediastinal stoma after total removal of larynx and trachea. AB - In a 54-year-old woman the entire trachea and larynx were removed because of malignant tumor. The first attempt to construct a mediastinal tracheostoma with 1.5 cm left of the distal trachea failed due to stomal infection with aortic wall necrosis and bleeding. Dividing the ascending aorta mobilized the left main bronchus and allowed a stoma at the level of the carina to be constructed without tension. The aortic continuity was restored by a vascular prosthesis from ascending to abdominal aorta. The patient survived and the stoma healed primarily. One year postoperatively the patient is doing well without signs of recurrent tumor. PMID- 7495594 TI - Preventive medicine: time for another Boston Tea Party. PMID- 7495593 TI - Nocardia asteroides purulent pericarditis. AB - A case is reported of a 71-year-old woman who developed cardiac tamponade as a result of a pyopericardium. The causative organism was Nocardia asteroides. This is the first reported case in the United Kingdom and only the 9th to be reported in the world. Despite prior radiological and echocardiographic investigations, we were unprepared at surgery for the volume of pus encountered. The dense adhesions, adherent pericardium and loculations encountered emphasizes the need to explore such patients surgically. We would advocate vigilance as to the possibility of pericardial involvement, in any patient contracting Nocardia asteroides infection. The medical and surgical management of this case is discussed together with a review of the literature. PMID- 7495595 TI - Responsiveness of auto makers to a simple public health initiative. PMID- 7495596 TI - Raw shellfish consumption in California: the 1992 California Behavioral Risk Factor Survey. AB - We used the 1992 California Behavioral Risk Factor Surveillance System to study the prevalence of raw shellfish consumption in California and the demographic and behavioral characteristics of raw shellfish consumers. We used the logistic regression analysis of the weighted survey data with PC SAS and SUDAAN to adjust for the effects of age and gender. Twenty-three percent of the respondents in the survey reported that they ate raw shellfish; one third of these reported eating raw shellfish once a month. Higher prevalences of raw shellfish consumption were reported by men, persons 18-49 years old, those with income above $25,000 and education beyond high school than by women, individuals older than 49 years, and those with an income of $25,000 or less per year and 12 or fewer years of school. A higher percentage of persons with liver disease, stomach surgery, and a history of chronic alcohol drinking reported consumption of raw shellfish than did individuals without liver disease, previous stomach surgery, or a history of alcohol abuse. After adjustment for gender and age, those who reported acute (P < .01) and chronic (P < .01) drinking and driving while intoxicated (P < .01) were more likely to report consumption of raw shellfish. Two variables (lack of seat belt usage [P = 2] and cigarette smoking [P = .13]) were not significantly associated statistically with raw shellfish consumption. PMID- 7495597 TI - Breast cancer screening behaviors and intentions among asymptomatic women 50 years of age and older. AB - We examined predictors of (1) compliance with yearly mammography and clinical breast examination (CBE) and (2) intention to obtain mammography and CBE within the following year. Subjects were 312 asymptomatic female hospital employees, 50 years or older, who had participated in a free worksite breast cancer screening program. Mammograms and CBEs had been obtained by 79% and 76% of the subjects, respectively, during the preceding 12 months. The majority of the subjects indicated that they were very/extremely likely to obtain a mammogram and CBE in the next 12 months (88% and 87%, respectively). Multivariate analyses showed that perceived barriers to and physician recommendation of mammography were the strongest predictors of both breast cancer screening behaviors and intentions. Recency of participation in the educational component of the screening program was related to both compliance with mammography in the past 12 months and intention to obtain CBE in the next year. Having a first-degree female relative with a history of breast cancer was related to compliance with CBE in the past 12 months. Perceived efficacy of mammography was related to intention to obtain both mammography and CBE in the next year, and number of reasons for mammography was related to intention to obtain mammography in the next year. Our results suggest that physicians play a key role in motivating women to comply with breast cancer screening and that reducing barriers to obtaining mammography may increase use further. PMID- 7495598 TI - Prevalence and detection of HIV risk behavior in primary care: implications for clinical preventive services. AB - Despite their potential to reduce the incidence of HIV infection through primary prevention, family physicians report low levels of routine identification of patients at risk and counseling. This may reflect perceptions that few of their patients are at risk, that patients at risk will self-disclose during consultations, or that a physician-initiated approach is unacceptable to patients presenting for non-HIV-related problems. Our aim was to determine the prevalence of risk factors for HIV infection and HIV testing among patients in general practice, the acceptability to patients of opportunistic identification of risk during routine consultations and the accuracy of general practitioners' assessment of HIV risk. Our setting included randomly selected general practitioners' surgeries in metropolitan Sydney, Australia. We conducted a self administered survey about risk factors in a consecutive sample of patients 18-50 years of age and compared it to a checklist about patient's risk factors completed by general practitioners unaware of the patients' answers. Of 1,030 patients, 43 (4%) were at risk of HIV infection having received blood transfusions between 1980 and 1985 and seven (1%) had injected intravenous drugs in the previous 12 months. In the previous 12 months, 133 (21%) female and 110 (28%) male patients had been in nonmutually monogamous heterosexual relationships. Only 26% always used condoms. While the majority of patients indicated they were heterosexual, 42 (4%) were homosexual and 23 (2%) bisexual. Of those men who had had sex with other men, 39% always used condoms.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7495599 TI - Patterns of communication about AIDS among Hispanic and Anglo adolescents. AB - This study examined exposure to AIDS information from mass media and interpersonal communication among a nonprobability sample of Hispanic and Anglo lower middle-class adolescents. Subjects were drawn predominantly from community health clinics, word of mouth recruiting, public service announcements, churches, schools, and health fairs in San Diego County. Both Hispanic (n = 220) and Anglo (n = 159) youths reported substantial exposure to information about AIDS from both mass media and interpersonal communication. Hispanics watched more general television than Anglos, but had less exposure to newspapers and interpersonal communication with friends. Communication among friends was most consistently related to sociodemographic variables, with older, higher status persons and girls communicating to the greatest extent. Boys reported the greatest communication with family. Anglo youths had greater exposure to information about AIDS and condoms regardless of the medium of communication, while Hispanic adolescents had greater exposure to information on risks of IV drug use. Of the three main types of mass media, print and radio provided the most exposure to AIDS information. It is important for preventive medicine practitioners to exploit these differences in communication patterns when planning preventive intervention strategies that target specific adolescent populations. PMID- 7495600 TI - Cross-sectional relationship between diet and physical activity in two southeastern New England communities. AB - The hypothesis that health promotive diets associated with higher levels of habitual physical activity confound the relationship between regular physical activity and health has not been well explored in epidemiologic studies. We evaluated self-reported physical activity, Willett Food Frequency dietary data, sociodemographic and physiologic factors cross-sectionally for 2,004 household survey participants in two southeastern New England communities. We compared the dietary habits of sedentary participants (n = 964) to those of moderately active (n = 600) and very active (n = 440) participants after adjusting for age, gender, education, smoking status, year of survey, and total calories. Our results showed that moderately active and very active participants consumed more fiber, less total fat, and less saturated fat than sedentary participants (P < .01). They also consumed more vitamins (A, C, D, E), beta carotene, and calcium, (P < .01), and ate more fruits and vegetables (P < .001) than sedentary participants. We found these relationships in both New England communities studied. This association between regular physical activity and diet suggests that the relationship of habitual physical activity and chronic disease may be confounded by diet. PMID- 7495601 TI - The association between veteran status and cigarette-smoking behaviors. AB - Although the prevalence of smoking has decreased since 1980 among active duty military personnel, it remains higher than among the adult civilian population; among military veterans, the prevalence of smoking has not been well described. The objectives of this study were to describe patterns of cigarette smoking behaviors among United States veterans and nonveterans and to examine the association between military veteran status and cigarette smoking. We analyzed data from a cross-sectional survey from a national probability sample of the civilian, noninstitutionalized adult population (National Health Interview Survey supplements). We estimated the prevalence of ever, current, and former smoking, as well as crude and adjusted odds ratios (AORs) of each outcome measure among veterans and nonveterans, by gender. The prevalence of ever smoking was 74.2% (+/ 0.7%) among veterans and 48.4% (+/- 0.5%) among nonveterans; current smoking prevalence was 33.9% (+/- 1.0%) among veterans and 27.7% (+/- 0.5%) among nonveterans. Among those who had not initiated smoking before the age of 18 years, veterans were more likely than nonveterans to report ever smoking (AOR = 1.8 for men and 1.9 for women) and current smoking (AOR = 1.9 for both men and women). After statistical adjustment, no difference was seen in cessation behavior. We concluded that the prevalence of ever and current smoking was higher among U.S. military veterans. The association was the strongest among veterans who had not initiated smoking before the age of 18 years. These findings are consistent with the hypothesis that military service is a risk factor for cigarette smoking, and they support the military's current prevention and cessation efforts. PMID- 7495602 TI - Ethnic intermarriage and its consequences for cystic fibrosis carrier screening. AB - Cystic fibrosis gene mutations can vary in frequency between different ethnic populations. However, there is a rising trend of ethnic intermarriage in the United States, a situation suggesting that differences in specific mutation frequencies currently apparent in Europe may not persist for long in this country. Therefore, limited mutation screens targeted at specific ethnic groups and risk calculations based on data from more homogeneous European populations may not be appropriate in the United States. The genetic consequences of ethnic admixture are also likely to extend to other recessive diseases (e.g., Tay-Sachs, thalassemia), which, in the past, have been limited largely to particular ethnic, racial, or religious subgroups, with implications for public health agencies overseeing newborn screening programs for genetic diseases and for clinical genetics programs offering population-based carrier-detection programs, carrier risk assessment, and counseling. PMID- 7495603 TI - Socioeconomic status and preventive health-care use by children in Spain. AB - National health systems are intended to provide equal access to health-care services to whole populations. However, they do not seem to address successfully the problem of social class differentials in access to health care, in particular access to preventive care. This study examines the relationship between the socioeconomic status (SES) of families and the use of preventive health care by children under a national health system in Spain. The study is based on weighted multivariate ordinal logistic regression analyses of data from the 1987 Spanish National Health Survey for a sample of 5,622 children, one to 10 years of age. A positive relationship was found between preventive health-care use by children and the SES of their families. Adult respondents' level of education and total family income were the most influential variables in this relationship. As these increased, children were more likely to receive visual, hearing, and dental exams. In addition, there was a gradient effect between family income and rate at which children received these preventive health-care services. Universal access to care, like that available in Spain, does not guarantee that social inequalities in children's receipt of preventive health care will not persist. In order to succeed, health-care reform must deal with social issues beyond financial access to care. PMID- 7495604 TI - Do power frequency magnetic fields cause leukemia in children? AB - The possibility of a causal relationship between exposure to electromagnetic fields and childhood leukemia has been studied for more than 10 years. This review evaluates the epidemiologic findings relevant to a possible causal association between exposure to power frequency magnetic fields during childhood and leukemia. The nine published studies on this topic were analyzed, with a focus on the selection of subjects, assessment of exposure, and control of confounding. Consistency of a positive association and dose-response relationship are evident for assessment of past exposure: among the five recent studies, relative risk estimates vary from 1.5 to 2.7 for past exposure assessment, and a significant dose-response relation was found in three studies. These observations cannot be explained by misclassification of exposure or consistent selection bias, and a confounder effect could hardly explain such results. Even if there is no currently understood biological plausibility for such an association, its possible causal nature cannot be dismissed. The impact on public health of such a possible causal association is difficult to assess precisely but could be significant. Therefore, it might be prudent to consider interim preventive measures while we await results of further research. PMID- 7495606 TI - Findings of scientific misconduct. PMID- 7495607 TI - Findings of scientific misconduct. PMID- 7495608 TI - [Problematic points of psychotropic drugs]. PMID- 7495605 TI - Influenza vaccine use among an elderly population in a community intervention. AB - A community intervention program was carried out to promote influenza immunization among an elderly population, as part of a demonstration established to estimate the value of including influenza immunization as a Medicare-covered benefit. Potential recipients were informed of reasons for receiving vaccine and its availability at no cost, and physicians and others administering it were provided with educational materials and cost reimbursement. Data collected for annual evaluations of influenza vaccine effectiveness were used to evaluate the impact of the community intervention program on influenza immunization status and to determine the effects of age, gender, chronic health conditions, and smoking status on the likelihood of immunization. Results indicated that the intervention program had a statistically significant impact on increasing the likelihood of immunization among the elderly population living in the targeted area. The existence of certain chronic health conditions also predicted immunization, while the effect of current smoking on immunization was strongly negative. During the three years of vaccine promotion, use in those 65 years and older who resided in the intervention area increased from approximately 40% in 1989-1990 to over 56% in 1991-1992. With various incentives, including cost reimbursement through Medicare, influenza immunization can be markedly increased among elderly beneficiaries. PMID- 7495609 TI - [Free radicals and neurological disorders]. PMID- 7495610 TI - [Assessment for intelligence on patients with Pick's disease]. AB - In contrast to Alzheimer's disease, it is unclear whether the MMS (Mini-Mental State) or the RCPM (Raven's Coloured Progressive Matrices) is clinically useful as a mental test battery in patients with Pick's disease, and it has not been fully evaluated whether intelligence of these patients really deteriorate along with the staging of dementia. The present study aimed to make these problems clear. We administered the MMS and the RCPM to 17 cases of Pick's disease, including 7 cases with predominantly frontal lobe atrophy and 10 cases with predominantly temporal lobe atrophy, which were all classified into 3 groups according to the staging of dementia by using the NM scale (New Clinical Scale for Rating of Mental States). We also investigated whether these patients had "Denkfaulheit" (laziness of thinking) and/or "gogi" aphasia (word meaning aphasia), which may affect performances on the mental test batteries. Two cases with predominantly frontal lobe atrophy and 5 cases with predominantly temporal lobe atrophy were assessed by the same examinations several years later. In cases with predominantly frontal lobe atrophy, all the patients presented with "Denkfaulheit". However, the mean RCPM score was normal in the mildly damaged patients, suggesting that their intelligence were relatively preserved. In the moderately and severely damaged groups, their intelligence could not be assessed owing to their severe "Denkfaulheit". In cases with predominantly temporal lobe atrophy, all the patients had "gogi" aphasia and 3 cases of the severely damaged groups presented with "Denkfaulheit".(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7495611 TI - [Comparison between anomia with word comprehension difficulty and Gogi aphasia due to lobar atrophy]. AB - We used naming and pointing tests and a proverb completion task to compare two cases of aphasia that exhibited a selective disturbance of word processing. One patient had anomia with word comprehension difficulty due to partial ablation of the left temporal lobe and the other patient had Gogi aphasia due to lobar atrophy with left temporal predominance. We presented 90 pictures of common objects divided into 9 categories in the naming and pointing tests, and used 10 well-known Japanese proverbs as stimuli in the proverb completion task. Performance of the naming and pointing tests was severely impaired in both patients. In the patient with anomia, words the patient could not name or point to varied from session to session and phonemic cue effects were frequently observed. The proverb completion phenomenon was positive. These findings indicate that the patient had an obstruction of the access route to the intact word store or a partial rarefaction of the word store itself. In the patient with Gogi aphasia, the words the patient could not name or point to were consistent from one occasion to another, and no phonemic cue effects or signs of familiarity were observed at all. The proverb completion phenomenon was totally negative. These findings indicate that the patient has lost the word store itself. MR images in the case of anomia revealed a lesion extending from the anterior to the central portion of the inferior part of the left temporal lobe. In the case of Gogi aphasia, the MR images displayed knife-edged focal atrophy in the anterior aspect of both temporal lobes, more prominently on the left.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7495612 TI - [Cerebellin in the cerebellum in spinocerebellar degeneration]. AB - To clarify the possible pathophysiological role of cerebellin in spinocerebellar degeneration, immunoreactive cerebellin was measured in the postmortem brain of 4 patients with spinocerebellar degeneration and 4 controls. Three other representative neuropeptides, corticotropin releasing hormone (CRH), neuropeptide Y and somatostatin, were also measured. Significant decreases in the concentration of immunoreactive cerebellin and immunoreactive CRH were found in the cerebellar hemisphere in spinocerebellar degeneration, suggesting an important pathophysiological role of cerebellin and CRH in this group of diseases. No such decreases were found in neuropeptide Y or somatostatin. PMID- 7495613 TI - [Histopathological analysis of herniated cerebellar tonsils resected from the patients with Chiari type I malformation with syringomyelia]. AB - Cerebellar tonsillectomy is a controversial treatment for Chiari type I malformation combined with syringomyelia. To demonstrate the validity of this procedure, we evaluated the postoperative clinical course, the histopathological finding in the resected cerebellar tonsils and the MR images of four patients. The patients were all women, ranging in age from 27 to 58 years old (mean: 43 years). The postoperative follows-up period lasted from 28 to 51 months (average: 38 mos), and the neurological symptoms and signs improved in all patients. Histopathological examination of the resected tonsils revealed a loss of Purkinje cells and granule cells. Vacuolated degeneration and chromatolysis were also seen, and modified Bielschowsky stain revealed axonal degeneration. These findings appeared to be irreversible. Postoperative T1-weighted MR images of the cervical and thoracic spine demonstrated a decrease in the size of the syrinx and the disappearance of evidence of tonsillar herniation in all patients. Postoperative phase-contrast MR images showed good CSF pulsation in the subarachnoid space at the craniocervical junction in all three patients examined. It was possible to avoid surgical complications by using careful microsurgical techniques. Based on these results, we concluded that cerebellar tonsillectomy is an adequate surgical strategies for treating Chiari type I malformation associated with syringomyelia. PMID- 7495614 TI - [Auditory comprehension in transcortical motor aphasia due to a medial lesions of the left frontal lobe]. AB - We assessed the anatomical findings and auditory comprehension of six patients with transcortical motor aphasia due to medical lesions of the left frontal lobe. All patients were right-handed and were initially mute for several hours after the onset, and they exhibited mild paresis of the right lower extremity. Their spontaneous speech was sparse and not fluent, and sometimes accompanied by echolalia, but their articulation was normal and repetition was excellent. They had difficulty in recalling words. A diagnosis of transcortical motor aphasia was made on the basis of their clinical symptoms. All patients were found to have an infarct in the left medial frontal region by MRI and/or CT. We administered the Western Aphasia Battery and 50 line drawing pointing task in order to evaluate auditory comprehension. Based on the results we concluded that there is no impairment of auditory comprehension of single words when lesions are limited to the superior frontal gyrus, but that lesions extending to the middle frontal gyrus interfere with auditory comprehension of single words. Our observations indicate that the middle frontal gyrus plays an important role in auditory comprehension of single words. All of the patients displayed impaired auditory comprehension of sentences even when their lesions were strictly limited to the medial frontal lobe. This suggests that the medial frontal lobe plays some role in the auditory comprehension of sentences. PMID- 7495615 TI - [A case of cranial metastasis of hepatocellular carcinoma]. AB - A case of cranial metastasis of hepatocellular carcinoma is reported. A 77-year old woman with an elastic hard tumor in the right temporal region was referred to our department on April 30, 1992. On admission, the patient had slight weakness of the left upper limb. Plain skull X-ray and computed tomography (CT) showed bone destruction in the right temporal region. Magnetic resonance images (MRI) showed that the tumor was hypo-intense with T1-sequences and hyper-intense with T2-sequences, and included hyper-intense spots on both T1- and T2-images. Right carotid angiography showed that the tumor was fed by the middle meningeal and accessory meningeal arteries. The patient became disoriented, and the left hemiparesis worsened on May 4, 1992. CT scan revealed an irregular high-density area in the tumor because of intratumoral hemorrhage. After embolization of the arteries feeding the tumor, surgery was performed on May 8, 1992, and the tumor was totally removed. Histological examination of the tumor specimen revealed that it was a metastatic hepatocellular carcinoma. The patient died 8 months after initial diagnosis because of hepatocellular carcinoma. PMID- 7495616 TI - [Cystic ectopic pituitary adenoma: report of a case]. AB - A 68-year-old man was admitted with suprasellar cystic tumor with obstructive hydrocephalus, and complaining of slowly progressive memory disturbance, gait disturbance and urinary incontinence. Neurological examination revealed bilateral visual disturbance and disorientation. Skull X-ray films revealed no dilatation of sella turnica and mild erosion of the dorsum sellae. A cystic suprasellar tumor was revealed extending upward to the third ventricular floor and the solid part of the tumor was homogeneously enhanced with Gd-DTPA on MRI. The T1-weighted sagittal MR image revealed a normal pituitary gland and the diaphragma sellae below the tumor. The tumor was totally removed via a bifrontal basal interhemispheric approach, and the tumor was attached to the pituitary stalk and was not contiguous with the pituitary gland. The histological diagnosis was sinusoidal type of chromophobe and non-functioning adenoma. Postoperative MRI revealed a preserved normal pituitary gland and the stalk. To our knowledge, only 11 cases of suprasellar ectopic pituitary adenoma have been reported. We reviewed their neuroradiological features and discussed the pathogenesis. In particular, cystic and nonfunctioning suprasellar ectopic pituitary adenoma may be difficult to distinguish from craniopharyngioma. PMID- 7495618 TI - [Persistent carotid-superior cerebellar artery anastomosis presenting with cerebellar ischemic attack: a case of persistent trigeminal artery variant]. AB - A rare case of persistent primitive trigeminal artery variant (PTAV) with cerebellar ischemia is reported. A 23-year-old male complained of sudden dizziness and nausea after playing valley ball. CT scan and MRI on admission revealed no abnormal findings. Left carotid angiography demonstrated a PTAV anastomoting precavernous portion of left internal carotid artery to the left superior cerebellar artery. The 37 cases reported in literature were reviewed to characterise PTAV. Ninety-seven% of the cases arising from precavernous portion of internal carotid artery, and terminated in anterior inferior cerebellar artery in 73%, posterior inferior cerebellar artery in 13.5% and superior cerebellar artery in 13.5%. Approximately 22.2% of patients with PTAV have cerebral aneurysms. The hypotension or mechanical compression of PTAV on playing valley ball with poor vascular supply to the part of cerebellum possibly caused cerebellar ischemia in this case. PMID- 7495617 TI - [A case of idiopathic hypoparathyroidism with slurred speech and mild dementia]. AB - A case of idiopathic hypoparathyroidism (IHP) is discussed with reference to a study of rats maintained on various low-calcium diets. A 45-year-old man complained of difficulty speaking, especially an inability to speak loudly and of fine finger tremors over the past 2 or 3 years. His brain MRI showed marked calcification in the bilateral basal ganglia and cerebellum and his in sera findings showed low calcium, extremely low levels in all kinds of examined parathyroid hormones and 1,25 (OH)2D, thus we diagnosed idiopathic hypoparathyroidism. His finger tremors stopped soon after administration of 1 alpha D3 2 microgram/day as verified by calcium, phosphorus and creatinine levels in sera, however, a brain CT one year later showed no changes in calcification. As reported earlier in studies of deposition in the soft tissues, especially central nervous system tissues of rats maintained on low-calcium diets, it is difficult to reduce calcium deposition if symptoms appear in adults. Therefore, it is necessary to start therapy as soon as possible in order to prevent additional calcium deposition in the early stage of IHP. PMID- 7495619 TI - [A 49-year-old man with progressive dysarthria, dysphagia, and left hemiparesis]. AB - We report a 49-year-old man who presented progressive dysarthria, dysphagia, and left hemiparesis. The patient was well until June 28th of 1993 when he noted 'weakness' in his both legs; despite his weakness, he could play golf on that day. In the beginning of July, he noted difficulty in swallowing solid foods. He was admitted to the neurosurgery service of our hospital on July 15th of 1993 and a neurologic consultation was asked on July 17th. Neurologic examination at that time revealed an alert but somewhat childish man who appeared to have some difficulty in paying attention to questions. He was disoriented to time and showed difficulty in recent memory and calculation. Higher cerebral functions were intact. The optic fundi were normal; pupils were isocoric and reacted to light promptly; ocular movements were intact, however, he showed difficulty in convergence. Facial sensation and facial muscles were intact. He had no deafness. He showed slurred speech and difficulty in swallowing solid foods. The remaining cranial nerves were intact. Motor-wise, he was able to walk normally and no weakness or atrophy was noted. Mild ataxia was noted in the finger-to-nose and the heel-to-knee test on the left. Muscle stretch reflexes were normal and symmetric, however, the plantar response was extensor bilaterally. Sensation was intact and no meningeal signs were noted. General routine laboratory findings were unremarkable. CSF was under a normal pressure containing 1 cell/microliter, 68 mg/dl of protein, and 54 mg/dl of glucose. Cranial CT scan showed low density areas involving the pons, midbrain, left thalamus, and the left parietal cortex. In MRI, these areas presented low signal intensity in T1-weighted images and high signal intensity in T2-weighted in images. The brain stem appeared swollen. Gadolinium enhancement was negative. He was given a course of steroid pulse with 1 g/day of DIV methylprednisolone for three days followed by oral steroid. He showed only temporary improvement in swallowing. In the subsequent course, he showed progressive deterioration in dysarthria and dysphagia. A biopsy was performed on the left parietal lobe lesion. After biopsy, he was treated with steroid and glycerol without improvement. A course of chemotherapy with procarbazine, MCNU, and vincristine was given; he did not respond to chemotherapy. His left hemiparesis deteriorated. He developed aspiration pneumonia from dysphagia and expired on October 22, 1993. The patient was discussed in a neurologic CPC, and the chief discussant arrived at the conclusion that the patient had astrocytoma grade III involving the pons, midbrain, thalamus, and the parietal cortex.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 7495620 TI - Want or need? PMID- 7495621 TI - MRI--the story so far. AB - On October 20, 1994, the Management Research Initiative was launched. Its aim? To support and publish sound quality research in management specifically related to dental practice so that a body of informed knowledge is available for dentists who want real answers to their questions about what management techniques work in practice. This article brings you up to date on what the MRI has achieved so far. PMID- 7495622 TI - Interview with Brian Gordon. Interview by Teresa Jane Waddington. PMID- 7495623 TI - Chronic sinusitis. PMID- 7495624 TI - The patient's view of minor oral surgery. PMID- 7495625 TI - Disinfection of impressions. PMID- 7495626 TI - Radiological standards for primary dental care. PMID- 7495627 TI - Struck off the dentists register. PMID- 7495628 TI - Prosthetic management of the atrophic mandible using endosseous implants and overdentures: a six year review. AB - This paper presents the treatment results and experiences gained from a retrospective study of patients treated with the IMZ osseo-integrated implant system and mandibular overdentures. Patients experiencing problems wearing conventional dentures were assessed by the implant team and 65 cases were treated with 154 endosseous implants placed in the edentulous mandible. Two to four implants were placed in each case and in addition some patients have had augmentation of the mandible with hydroxyapatite. The definitive mandibular prostheses were supported by both implants and the residual ridges. A variety of retention systems were utilised, which included different types of bar and clip, stud attachments, and magnets. The patients have been followed up regularly and evaluated after periods of between one and six years. Six implants have failed over this time resulting in a success rate of over 96%. Most patients expressed a high degree of satisfaction with their new overdentures. There was, however, a considerable burden of maintenance care required for the patient group examined. The findings demonstrate that implant retained overdentures offer a highly effective means of oral rehabilitation for the atrophic mandible, restoring both oral function and facial form. PMID- 7495629 TI - Prevention and detection of oral cancer: the views of primary care dentists in Northern Ireland. AB - Our objective was to describe the management of oral cancer and pre-cancer as stated by primary care dentists and their views on screening. We conducted a survey of all general dental practitioners and community dentists in Northern Ireland (n = 635), to which 428 replied (response rate: 67%). 94% stated that examination of the oral soft tissues constituted part of their usual practice during the regular dental check-up. Suspicious lesions were generally referred early, 68.5% of dentists referring white lesions within one month of presentation. The corresponding figures for red lesions, lumps and persistent ulcers were 80.1%, 89.7% and 91.7%. The incidence of oral cancer was over estimated (median '70' cases/year, versus the true figure of approximately 40/year) as, in all likelihood, was the percentage by which mortality could be reduced by screening (median: 50%). Accordingly the adoption of a screening programme was favoured over investment in health promotion. Indeed, only 14% said that their patients records routinely contained information about smoking or alcohol habits. Although there are some areas of practice which could improve and the potential of screening is probably over-valued, primary care dentists in Northern Ireland already opportunistically screen and refer patients promptly. PMID- 7495630 TI - Direct composite inlays versus conventional composite restorations: three-year clinical results. AB - A clinical trial of direct composite inlays versus conventionally (incrementally) placed restorations made from the same material was started in January 1989. Restorations were randomly allocated to matched pairs of cavities. This study reports the 3-year performance of 71 of the 100 pairs of restorations placed over a 2-year period and followed-up every 6 months. Clinical assessments were made using USPHS criteria (indirect measurements of occlusal wear were made using Ivoclar standard dies) and annual bite wing radiographs. Direct inlays showed significantly less occlusal wear than conventional restorations, but the difference was small. The clinical performance of both types of restoration was similar and compared favourably with studies of other materials. No secondary decay was diagnosed. The direct inlays, however, took longer to place and did not reduce postoperative sensitivity or failure rate (8% failure of inlays and 4% of conventional composites over 3 years). Contouring of proximal and occlusal aspects was not facilitated with direct inlays but may be easier with indirect inlays on removable dies. PMID- 7495631 TI - General dental practice--challenges and opportunities: a personal view. AB - This second paper in the Future of Dentistry series considers the implications for general dental practice of trends in health care nationally and internationally. Its main objective is to increase awareness of the major changes occurring in primary care which provide the dental profession with new challenges and opportunities. It is important that the dental profession is given its proper place in the changing scene and that general dental practice is not isolated from new developments in primary health care. PMID- 7495632 TI - The role of the shortened dental arch concept in the management of reduced dentitions. AB - Current research indicates that a more biological approach to dental care is required to cope with current trends in adult health and to make more efficient use of ever dwindling public financial resources. This paper describes the role of the shortened dental arch concept (SDA) in the management of reduced dentitions, including criteria for its application, possible indications and contraindications, and presents a review of research supporting SDA. In light of recent research, it is suggested that SDA will be of increasing importance as a treatment philosophy for the future elderly. PMID- 7495633 TI - Creating the story of a life. PMID- 7495634 TI - Hospice RN's guide to chemotherapeutic agents. PMID- 7495635 TI - Palliative care for patients with acquired immunodeficiency syndrome. AB - This article provides a clinically-oriented overview of palliative care for patients with AIDS. Indicators of decreased survival time are divided into categories of infections/illnesses, clinical signs and symptoms, immunological and serological markers, and psychosocial factors. Primary symptoms in AIDS are discussed according to etiology and treatment. However, treatments of opportunistic infections per se are not directly addressed in this article. Problems discussed include pain, confusion, depression and anxiety, fatigue, fever, dyspnea, nausea and vomiting, diarrhea, wasting, and dehydration. The article also briefly addresses clinical and ethical questions and challenges presented by AIDS to hospice or palliative care providers, and the various stages of HIV infection. PMID- 7495636 TI - Physician assisted suicide & voluntary active euthanasia: social ethics and the role of hospice. PMID- 7495637 TI - Marketing hospice. PMID- 7495639 TI - Developing family care plans: a systems perspective for helping hospice families. AB - Although terminal care literature may address the importance of the family in the patient's treatment, the family as a whole is seldom considered as the unit of care. A family system's perspective can offer a more holistic approach to providing care to the terminally ill. In this paper, Rosen's model of family system response to stress is adapted to provide a framework for understanding and helping hospice families. To ascertain information critical to the development of a comprehensive care plan, a general format is outlined. Case examples illustrate the application of this approach. PMID- 7495638 TI - Beyond DNR: a consideration of four cases. PMID- 7495640 TI - Stereoselectivity of the carbonyl reduction of dolasetron in rats, dogs, and humans. AB - The initial step in the metabolism of dolasetron or MDL 73,147EF [(2 alpha, 6 alpha, 8 alpha, 9a beta)-octahydro-3-oxo-2,6-methano-2H- quinolizin-8-yl 1H-indol 3-carboxylate, monomethanesulfonate] is the reduction of the prochiral carbonyl group to give a chiral secondary alcohol "reduced dolasetron." An HPLC method, using a chiral column to separate reduced dolasetron enantiomers, has been developed and used to measure enantiomers in urine of rats, dogs, and humans after dolasetron administration. In all cases, the reduction was enantioselective for the (+)-(R)-enantiomer, although the dog showed lower stereoselectivity, especially after iv administration. An approximate enantiomeric ratio (+/-) of 90:10 was found in rat and human urine. The contribution of further metabolism to this enantiomeric ratio was considered small as preliminary studies showed that oxidation of the enantiomeric alcohols by human liver microsomes demonstrated only minor stereoselectivity. Further evidence for the role of stereoselective reduction in man was obtained from in vitro studies, where dolasetron was incubated with human whole blood. The enantiomeric composition of reduced dolasetron formed in human whole blood was the same as that found in human urine after administration of dolasetron. Enantioselectivity was not due to differences in the absorption, distribution, metabolism, or excretion of enantiomers, as iv or oral administration of rac-reduced dolasetron to rats and dogs lead to the recovery, in urine, of essentially the same enantiomeric composition as the dose administered. it is fortuitous that the (+)-(R)-enantiomer is predominantly formed by carbonyl reductase, as it is the more active compound. PMID- 7495641 TI - Comparative toxicity of (+)-(R)- and (-)-(S)-1,2-dibromo-3-chloropropane. AB - The haloalkane 1,2-dibromo-3-chloropropane (DBCP), an environmental pollutant that was widely used as a soil fumigant, is a carcinogen and a mutagen and displays target-organ toxicity to the testes and the kidneys. Because little is known about effects of stereochemistry on the metabolism and toxicity of halogenated alkyl compounds and because DBCP, which has a chiral center at C-2, may show enantioselectivity in its metabolism and/or toxicities, the optically pure enantiomers of DBCP were tested in vivo in rats for organ toxicity as well as for bacterial mutagenicity. Organ toxicity studies showed that (S)-DBCP was slightly more renal toxic than (R)-DBCP but was not significantly more toxic than the racemate, and that no significant differences were observed in the extents of testicular necrosis and atrophy caused by either enantiomer or the racemate. In contrast, (R)-DBCP was more mutagenic than either (S)-DBCP or the racemate to Salmonella typhimurium (S. typhimurium) strains TA 100 and TA104. However, there was little or no enantioselectivity in glutathione S-transferase (GST)-catalyzed conjugation reactions of glutathione with DBCP based on the lack of selectivity in the rates of disappearance of the enantiomers of DBCP in the presence of glutathione (GSH) and GSTs as monitored by chiral gas chromatography (GC). PMID- 7495642 TI - Inversion of (R)- to (S)-ketoprofen in eight animal species. AB - The (R)-enantiomer of the NSAID ketoprofen was administered orally at 20 mg/kg to a series of 8 animal species. In all species, a highly significant degree of inversion occurred after 1 h which varied from 27% (gerbil) to 73% (dog) and persisted or increased in plasma samples obtained 3 h after drug administration. Although the (R)-enantiomer was inactive as an inhibitor of cyclooxygenase, the analgesic effects of that isomer was almost the same as the (S)-isomer in animal analgesic assays, following oral administration of the drugs to mice and rats. Taken together, the present results suggest that (R)-ketoprofen administered alone functioned primarily as a prodrug for (S)-ketoprofen under the experimental conditions of this study. PMID- 7495644 TI - In vitro fenoprofenyl-coenzyme A thioester formation: interspecies variations. AB - In vitro coenzyme A thioester formation from (-)-(R)-fenoprofen (FPF) and palmitic acid has been studied using liver microsomes from rat, guinea pig, sheep, and dog. In every species with both palmitic acid or (-)-(R)-fenoprofen, the Lineweaver-Burk plot was linear in the substrate concentration range used and as a consequence agrees with the involvement of only one isoenzyme (or different isoenzymes of similar Km values). The Vmax values for the thioesterification of ( )-(R)-fenoprofen present large species variations from 2.1 +/- 1.0 with sheep liver microsomes to 60.6 +/- 11 nmol/min/mg with dog liver microsomes. These values statistically significantly correlate (r = 0.94) to the Vmax values observed when palmitic acid was used as a substrate. Furthermore palmitic acid inhibited (-)-(R)-fenoprofen-CoA formation in the same extent in all animal species. The stereoselectivity of the thioesterification was also species dependent. PMID- 7495643 TI - Absolute configurations and conformations of the opioid agonist and antagonist enantiomers of picenadol. AB - The absolute configurations of the enantiomers of the opioid picenadol [cis-1,3 dimethyl-4-propyl-4-(3-hydroxyphenyl)piperidine; cis-3-methyl, 4-propyl] have been determined by an X-ray crystallographic study of the chloride salt of the (+)-enantiomer. The agonist (+)-enantiomer and the antagonist (-)-enantiomer were found to have the 3R,4R and 3S,4S absolute configurations, respectively. The conformational properties of the enantiomers were also examined with MM2-87 calculations. There was good agreement between the computed global minimum and the crystallographic structure with the phenyl ring approximately bisecting the piperidine ring by both methods. This orientation of the phenyl ring differs from that of related opioids such as the phenylmorphans, prodines, meperidine, and ketobemidone in which the phenyl ring tends to eclipse one edge of the piperidine ring. Because the phenyl ring bisects the piperidine ring in picenadol, there is little difference in the three-dimensional orientations of the phenyl rings of the two enantiomers when one superimposes the piperidine rings. The agonist (+) enantiomer is ambiguous with respect to an opioid ligand model, which suggests that agonist activity requires a specific range of dihedral angles for the phenyl ring. While the global minimum of the agonist is not consistent with the model, a second conformer that is only 1.2 kcal/mol above the global minimum is consistent. An alternative explanation is that agonist or antagonist activity is solely due to the presence of the 3-methyl group on the different edges of the piperidine ring.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7495645 TI - Meningococcal disease. PMID- 7495646 TI - Outbreak of Venezuelan equine encephalitis in Colombia and Venezuela. PMID- 7495647 TI - Expanded programme on immunization. Measles, 1994. PMID- 7495648 TI - Zoonoses control. Oral immunization of foxes in Europe in 1995. PMID- 7495649 TI - Disabilities among children up to 17 years of age 1991-1992. PMID- 7495650 TI - Demographic and health survey in Kenya, 1993. Maternal and child health. PMID- 7495651 TI - [Nursing and care of a child with nephrotic syndrome--from birth to transplantation]. PMID- 7495652 TI - [Care after kidney transplantation in children]. PMID- 7495653 TI - [Sedation and analgesia in neonatology. 1]. PMID- 7495654 TI - [Interaction-oriented help for premature babies and their parents]. PMID- 7495655 TI - [What to do at the suspicion of sexual child abuse?]. PMID- 7495657 TI - [Students are running a hospital department--a project of course 93/96 at the pediatric nursing school of the Freiburg University Hospital]. PMID- 7495656 TI - [Are temperature measurements in the auditory canal of newborns reliable?]. PMID- 7495658 TI - [Health for children in the year 2000--report on the press conference on September 11, 1995 on occasion of the 17th meeting of German pediatric nurses in Krefeld]. PMID- 7495659 TI - [Discrimination against male pediatric nurses?]. PMID- 7495660 TI - Primary diffuse chiasmatic germinomas: differentiation from optic chiasm gliomas. AB - Primary germ cell tumors confined to the optic nerves and chiasm without suprasellar extension are uncommon. These tumors appear similar to chiasmatic gliomas on both computed tomography and magnetic resonance imaging, potentially resulting in treatment errors if the diagnosis is based on radiologic criteria alone. Unlike chiasmatic gliomas, suprasellar germinomas characteristically present with a clinical triad of endocrine abnormalities, diabetes insipidus, and visual complaints. We report the case of a 9-year-old boy who presented with a 5 month history of fatigue, 16-pound weight gain, polydipsia, polyuria, visual complaints, and intermittent headache. Imaging studies demonstrated findings consistent with a glioma of the chiasm with infiltration into the optic tracts. At surgery, the chiasm and optic tracts were diffusely enlarged with no other suprasellar abnormalities. Biopsy specimens were characteristic of germ cell tumor. Based on this result, the patient received a treatment regimen different from that used at our institution for chiasmatic gliomas. We feel it is imperative to biopsy chiasmatic lesions that radiologically appear to be gliomas if symptoms do not adhere to the classical clinical presentation. PMID- 7495661 TI - Model for the cost analysis of shunted hydrocephalic children. AB - This paper describes a model for forecasting the treatment costs for hydrocephalic patients with ventriculoperitoneal shunts. Modeling with institution-specific or reported failure rates allows the prediction of shunt failure in real and/or theoretical populations. The addition of costing factors (derived from hospitalization, operative and drug costs) to the model allows the derivation of partial or total cost estimates. The effects of varying the failure rate, infection rate, number of new patients, number of lost patients and costing factors can be simulated and measured. Basing this model on data from our institution, decreasing the rate of failure during the first year following shunt insertion or revision has the potential for greater cost savings over time than either decreasing the shunt infection rates or the duration of hospital stay. By combining shunt performance and financial data, an estimate of the cost of the treatment of a population with hydrocephalus, over time, can be derived. These data can be critical for institutional and program budgeting and serve as an estimate of the economic effects of treatment changes proposed in clinical trials. PMID- 7495662 TI - Evidence for a patent fibrous tract in fractured, outgrown, or disconnected ventriculoperitoneal shunts. AB - As cerebrospinal fluid (CSF) shunts function for several years, outgrowth and disconnection of the distal catheter are seen more frequently. Though a shunt revision is clearly indicated if patients are symptomatic, asymptomatic patients with a disconnection or who have outgrown the distal catheter may no longer require CSF diversion. This follows an assumption that the shunt is no longer functioning and, in an asymptomatic patient, no longer required. However, patent fibrous tracts have been postulated which may allow passage of CSF through disconnected or outgrown distal catheters. We have recently treated 2 patients with ventriculoperitoneal (VP) shunts with evidence of a persistent fibrous tract. A 13-year-old boy was asymptomatic for 2 years after radiographs revealed an outgrown distal VP shunt catheter, after which the acutely deteriorated and died from massive hydrocephalus, although the entire shunt was patent. Another 14 year-old boy who was profoundly retarded presented with subtle changes in his daily activity. Shunt radiographs revealed two disconnections and a broken valve, but a shunt tap revealed a normal opening pressure (OP). A radionuclide shuntogram revealed rapid passage of isotope through fibrous sheaths at the disconnections, and into the peritoneum. A shunt revision was performed and the patient returned to his previous level of health. Patients with outgrown or disconnected distal catheters may be asymptomatic and/or have normal OP, and must not be assumed to have arrested hydrocephalus. A ventricular pressure measurement and radionuclide shuntogram should be performed to rule out a patent persistent fibrous tract. PMID- 7495663 TI - Pial synangiosis for moyamoya syndrome in children. AB - The authors describe a modification of the encephalo-duro-arterio-synangiosis (EDAS) surgical technique for patients with moyamoya syndrome in which the arachnoid is widely opened over the exposed brain and the donor superficial temporal artery adventitia is sutured directly to the pia with 10-O nylon suture ('pial synangiosis'). The procedure has been performed in 34 patients over a 10 year period, with follow-up arteriograms 1 year following the procedure demonstrating markedly enhanced middle cerebral artery flow through the surgical site. Operative morbidity has been limited to two postoperative strokes, both in patients who were neurologically unstable preoperatively; and the clinical stability of the operated patients has thus far been excellent. PMID- 7495664 TI - Outcome from pediatric head injury: on the nature of prospective and retrospective studies. 1989. PMID- 7495665 TI - Cerebrospinal fluid reabsorption. A critical appraisal. 1990. PMID- 7495666 TI - Illi intracranial pressure monitoring and fixation device. A new fixation system for intracranial pressure monitoring in children and adolescents in paediatric neurotraumatology, hydrocephalus and craniofacial surgery. AB - Fixation and positioning of intracranial pressure (ICP) monitoring devices in small children with very thin skull bones pose serious problems. We present a new ICP fixation device with two movable components compressing the skull bone from in- and outside. This new fixation system, designed by PD Dr. O.E. Illi and licensed by White Spot AG, Surgical Tools and Research, Baar, Switzerland, guarantees far greater stability and reliability than conventional screws or bolts. It is fully compatible with a range of fiberoptic or pneumatic transducer systems. We have applied this new device successfully in 10 patients. PMID- 7495667 TI - Value of dynamometry in assessing upper extremity function in children with myelomeningocele. AB - Children with myelodysplasia frequently have abnormal hand function. Etiologies include brainstem and cervical cord compression due to the Chiari II malformation, syringomyelia, hydrocephalus, and cerebral dysmorphism. Determination of potentially correctable causes of hand dysfunction may be difficult. We describe the use of the Jaymar dynamometer for assessing grip strength as a means of detecting early changes in hand function. PMID- 7495668 TI - Endothelin receptor antagonism. AB - Following the original report by Yanagisawa et al. (1988) more than 7 years ago, compelling evidence that ET plays an important role in the local regulation of smooth muscle tone and cell growth has been reported. In addition, many studies point to a significant role for endothelin in nonvascular function. The investigation of the endothelin system has been greatly advanced in the last 2 to 3 years through significant advances in the development of potent and selective ET receptor antagonists. These agents have proven to be essential tools for elucidating the biological significance of the ET system, leading to the realization that antagonism of the ET system may have significant therapeutic potential. As emphasized in this review, the importance of chronic blockade of the ET system may be a critical aspect of future research in this exciting area. Confounding issues remain the lack of information about the role of the ETB receptor, the apparent pharmacological evidence for additional ET receptor subtypes, and species variation in the tissue distribution of ET isoforms and receptor subtypes. Along with the greater ability to understand the endothelin system provided by potent and selective pharmacological agents, is the important contribution of modern molecular biology techniques, highlighted by the insights gained from recent reports of results from ET gene disruption studies. Kurihara et al. (1994) found that ET-1-deficient homozygous mice die at birth of apparent respiratory failure secondary to severe craniofacial abnormalities. Subsequently, Yanagisawa's laboratory has presented and published a series of complementary gene disruption studies. First, Hosoda et al. (1994) demonstrated remarkably, that ETA receptor knockout mice bear morphological abnormalities nearly identical to ET-1 knockout mice. Second, they found that disruption of the ET-3 peptide and ETB receptor genes result in homozygous mice that share identical phenotypic traits (i.e., coloration changes and aganglionic megacolon) which are similar to a previously known natural mutation, the Piebald-Lethal mouse (Hosoda et al., 1994; Baynash et al., 1994). This phenotype has a human corollary known as Hirschsprung's Disease and it is now known that the disease, though multigenic, results from a missense mutation of the ETB receptor gene in some individuals (Puffenberger et al., 1994). Taken together these data indicate that the endothelin system is essential to correct embryonic neural crest development, a completely novel finding within the superfamily of guanine-protein-linked receptors.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 7495669 TI - Alternative approaches for the application of ribozymes as gene therapies for retroviral infections. PMID- 7495671 TI - Regulation of airway wall remodeling: prospects for the development of novel antiasthma drugs. PMID- 7495670 TI - Inducible cyclooxygenase and nitric oxide synthase. PMID- 7495673 TI - Monoclonal antibody therapy of leukemia and lymphoma. PMID- 7495672 TI - Advances in selective immunosuppression. PMID- 7495674 TI - 4-Hydroxyphenylretinamide in the chemoprevention of cancer. AB - It has been suggested that ultimately half of all cancers might be prevented by early interventions (Costa, 1993). 4HPR has been shown to be an effective and safe agent in various in vivo animal trials and well tolerated in human trials. At present multiple clinical trials are assessing its efficacy in preventing a variety of cancers. PMID- 7495675 TI - Immunoconjugates and immunotoxins for therapy of carcinomas. PMID- 7495676 TI - Discovery and in vitro development of AIDS antiviral drugs as biopharmaceuticals. AB - The goal of developing an effective drug against HIV-1 and AIDS has been approached by several routes, with enough encouraging results to stimulate further efforts. Compounds active against HIV-1 have been discovered for many of the functions in the reproductive cycle recognized as virus-specific targets. Discoveries have been made in cell-based assays as well as mechanistic assays, and the value of both types of assays in the drug discovery process has been discussed. Although the final test of a drug's efficacy comes in the clinical experience, submission of an antiviral compound to an in vitro developmental gauntlet can save much time, effort, expense, and human resource in the in vivo developmental regimen required prior to human use. Emergence of viral resistance to drugs in several structural classes has compromised their clinical efficacy, suggesting that development of other potential drugs in those classes may not be good investments. Strains of HIV-1 resistant to specific compound classes are used to categorize new active discoveries for possible developmental exclusion, and defining the mechanism of action of such a new compound may confirm the discouraging judgement. On the other hand, novel compounds which exhibit a broad range of activity in drug-resistant and other HIV-1 strains deserve greater scrutiny. Clinicians most likely will be hesitant to treat patients with compounds shown to act on virus-cell surface interactions, given the failure in the past of several such compounds in clinical studies. But a compound shown to have a unique and novel mechanism of action will be looked upon more favorably, and surviving other tests of potency, solubility, and stability will be unhesitatingly presented for in vivo development. The partial successes of drugs currently in clinical use against AIDS offers great encouragement that other more effective, less-toxic drugs will be found. Exquisite techniques for identifying new targets on virus gene products, the selection of compounds on activity paradigms, and the enormous variety of compounds becoming available through synthesis libraries, all offer opportunities for anti-HIV drug discovery, which, in our view, cannot fail to present potent antiviral compounds which will survive the rigorous preclinical and clinical tests leading to a drug effective against AIDS. PMID- 7495677 TI - The ryanodine receptor family of intracellular calcium release channels. PMID- 7495678 TI - Design and pharmacology of peptide mimetics. PMID- 7495679 TI - Usefulness of myosin in the postmortem diagnosis of myocardial damage. AB - In some situations the postmortem diagnosis of myocardial infarction is made difficult by the brief course of the fatal episode or by interferences caused by autolysis. In such cases, biochemical indices may provide a useful adjunct to morphological studies. Myosin is the main component of the contractile apparatus of muscle cells, so its determination may well be useful to evaluate myocardial injury. The purpose of the present study was to establish the diagnostic efficacy of postmortem myosin heavy chain determinations using monoclonal antibodies and to compare this data with structural findings used to diagnose acute myocardial ischaemia. We studied 105 cadavers with a mean age of 61.63 +/- 2.21 years. Cases were allocated to 1 of 7 diagnostic groups depending on the probable intensity of myocardial damage and cause of death. The highest serum and pericardial fluid values of myosin heavy chains were seen in subjects who showed morphological evidence of myocardial ischaemia. Mean pericardial fluid/serum ratios differed significantly between subjects with and without observable signs of heart damage. PMID- 7495680 TI - Estimation of chronologic age using the aspartic acid racemization method. I. On human rib cartilage. AB - Determinations of chronologic age are of great importance in forensic science. At present the aspartic acid racemization method on teeth provides one of the best means in adult individuals. However, if teeth are not available, some other stable tissue has to be used. In this study, the applicability of cartilage from the ribs has been tested. Specimens of rib cartilage were obtained at autopsy from 24 individuals aged 0.2-96 years. An acid-soluble peptide (SP) fraction and an insoluble collagen-rich (IC) fraction were prepared, and the ratio of D/L aspartic acid was determined using the HPLC technique. The correlation coefficient between the D/L ratio and age was r = 0.91 in SP and r = 0.97 in IC. It thus seems as if cartilage from non-weight-bearing areas may be a useful source of tissue for the estimation of chronologic age with the aspartic acid racemization method when teeth are not available. PMID- 7495681 TI - Distribution of HLA DQA.1 alleles in New Zealand Caucasian, Maori and Pacific Islander populations. Comparison with other population studies. AB - Allele and genotype frequencies for the HLA DQA.1 locus were determined for 127 unrelated Caucasians, 177 unrelated Maori and 98 unrelated Pacific Islanders from the New Zealand population. DNA from blood cells was analysed by polymerase chain reaction amplification of DNA followed by hybridization to allele specific oligonucleotide probes in a reverse dot-blot test. Allele frequencies at the HLA DQA.1 locus for New Zealand Caucasians, Maori and Pacific Islanders were compared with published data for other populations. The distribution of HLA DQA.1 genotype frequencies did not deviate from Hardy Weinberg expectations for the Caucasian and Maori populations. The power of discrimination was 0.93 for Caucasians and 0.86 for Maori. The total Pacific Islander population tested was analysed as was data obtained from Western Polynesians contained within that larger group. Both the total Pacific Islander group analysed, and the Western Polynesians contained within that larger group, failed Hardy Weinberg expectations for the distribution of HLA DQA.1 genotypes. This significant deviation was due to excess homozygotes. The power of discrimination for the total Pacific Islander group and for Western Polynesians was 0.86 and 0.85 respectively. Comparison of Caucasian population studies from New Zealand, the United Kingdom, South Australia, Norway, the United States and Sweden showed these populations have similar HLA DQA.1 allele frequency distributions. Maori and Pacific Islanders have HLA DQA.1 allele frequency distributions that are more similar to each other than any of the other populations studied. PMID- 7495682 TI - Estimation of chronologic age using the aspartic acid racemization method. II. On human cortical bone. AB - Over the last 20 years a new chemical method, base on the racemization of aspartic acid, has been developed to be used for the estimation of chronologic age in adult individuals. The method has a good accuracy when used on dental enamel, dentine and cartilage. However, in forensic and archeological cases teeth and cartilage are not always available. Since preliminary studies have shown that there are some age-related changes of the D/L aspartic acid ratio also in bone, this study was carried out to further explore if the method could be used for age estimations of bone. Bone samples from 24 individuals, aged 0.2 to 95.6 years were analysed for the D/L ratios with HPLC-technique. Two different fractions of the bone were examined, an acid-soluble peptide fraction and an acid-insoluble collagen-rich fraction. The analyses showed age-related racemizations in both fractions, although of different rates. The correlation coefficients with age were 0.72 in the peptide fraction, and 0.84 in the collagen-rich fraction. It thus seems as if bone may be used for age estimations when more stable tissues like dentine and cartilage are not available. PMID- 7495683 TI - Immunohistochemical detection of early myocardial infarction. An evaluation of antibodies against the terminal complement complex (C5b-9). AB - Antibodies (abs) against the terminal complement complex (C5b-9) were used on routinely processed post mortem myocardial tissue in parallel with conventional staining methods. Both monoclonal and polyclonal abs were tested using the avidin biotin peroxidase complex (ABC), alkaline phosphatase anti-alkaline phosphatase (APAAP) methods and an ab-bridge with alkaline phosphatase. Enhancement of the diaminobenzene (DAB) end product with cobalt-nickel (ABC method) was also done. The polyclonal ab gave the most satisfactory results and the alkaline phosphate conjugated ab-bridge had a slight advantage over the ABC method. Cobalt-nickel enhancement of DAB improved the visualization, but with higher background staining. APAAP was the least satisfactory method. Comparing the immunohistochemical method with the conventional staining methods, the former showed positive reaction in 97% of areas of coagulation necrosis and in 65% of contraction band necrosis. On the other hand coagulation necrosis was seen in 44% and contraction band necrosis in 68% of C5b-9 positive areas indicating that C5b 9 abs react with ischemically damaged myocytes before visible alterations are seen in hematoxilin-eosin staining. Moreover, using C5b-9 abs, it seems possible to exclude agonal/artefactual contraction bands which show a negative reaction. Immunohistochemical detection of C5b-9, using an adequate technique could increase the possibility to demonstrate early ischemic myocardial damage. PMID- 7495684 TI - Structural variation of novel alleles at the Hum vWA and Hum FES/FPS short tandem repeat loci. AB - This paper reports the sequences of novel alleles identified during population databasing studies on the short tandem repeat loci HumvWA and HumFES/FPS. Two HumFES/FPS alleles follow the simple repeat pattern (ATTT)7 and (ATTT)15. Sequence variation corresponding to an A to C transversion occurred in the 5' flanking region in two individuals possessing the designated allele 7. Two HumvWA alleles exhibited compound repeat regions comprising TCTA and TCTG repeat units. Sequence analysis confirmed the putative designation of 11 for a 127 base pair allele. However, a 131 base pair allele, putatively designated as 12, exhibited a more complex sequence. Two different types of repeat unit structures were identified which also exhibited sequence variation in the 3' flanking region. PMID- 7495685 TI - Dutch Caucasian population data on the loci LDLR, GYPA, HBGG, D7S8, and GC. AB - To introduce the loci LDLR, GYPA, HBGG, D7S8, and GC (PM loci) in Dutch forensic identity testing, allele and genotype frequencies were determined in a Dutch Caucasian population sample, which had previously been typed for the HLADQA1 locus [12]. All 6 loci met Hardy-Weinberg equilibrium expectations, and there is little evidence for association between pairs of loci. The combined power of discrimination for all 6 loci is 0.9997. The allele frequencies of the PM loci were similar to 2 other Caucasian populations [3, 10], and differed from 3 non Caucasian populations [3]. PMID- 7495687 TI - Amplification and detection of the VNTR locus D4S95 in a Japanese population. AB - The D4S95-VNTR locus was amplified and the polymorphism analysed in a population sample of 169 randomly selected Japanese individuals. A total of 14 alleles containing 850-1360 base pairs were distinguished by agarose gel electrophoresis. The distribution of alleles was symmetrical with respect to one peak at 1030 bp. The mean exclusion chance and discrimination power were calculated as 0.604 and 0.876 respectively. PMID- 7495686 TI - Detection of green algae (Chlorophyceae) for the diagnosis of drowning. AB - The plankton test (generally, diatom test) is one of the methods available to diagnose the cause of death of submerged bodies. The solubilization method using tissue solubilizer Soluene-350 was used in this study to detect not only diatoms but also green algae, based on the fact that the solubilizer does not digest the cell walls of green algae which are made from cellulose. Detection of green algae from organs of submerged cadavers is very informative to determine drowning in fresh water, and also in cases where only few diatoms are detected in the organs. PMID- 7495688 TI - D1S80 (pMCT118): analysis of 3 ethnic subpopulations living in Brussels. AB - Population genetic studies were carried out on 3 ethnic subpopulations living in Brussels (119 Belgians, 120 Turks and 137 Moroccans). DNA extraction was performed using the Chelex method. After DNA amplification the DNA fragments were separated electrophoretically in horizontal polyacrylamide gels. A total of 32 alleles (between 21 and 25 alleles in each subpopulation) including 8 "interalleles" could be differentiated. The allele frequencies were compared with population data from a German study and no significant differences could be observed. PMID- 7495689 TI - Hungarian population data for the STR systems TH01 and VWA. AB - Population data studies for TH01 and VWA were carried out on a Hungarian caucasian population sample of 244 unrelated individuals. We found 7 and 8 different alleles for TH01 and VWA, respectively. Using a high resolution/non denaturing horizontal PAGE system no variations in the mobility of alleles were detected except for the allele 14 in VWA. The allele frequencies observed are similar to those reported in the literature for European caucasians. The genotype distributions meet Hardy-Weinberg expectations for both STR systems. PMID- 7495690 TI - Observations on the Lewis (a) and Lewis (b) activity of erythrocytes. PMID- 7495691 TI - Large scale database experiments to assess the significance of matching DNA profiles. AB - Over 5,700 three-probe VNTR DNA profiles collected by several United Kingdom (UK) laboratories have been compared to examine the probability of randomly matching 2 samples from different individuals. In over 16 million comparisons, using a matching rule corresponding to the matching guideline employed by the UK Forensic Science Service, no profiles were found to match at the 3 loci D1S7 (MS1), D7S21(MS31) and D12S11 (MS43a). The frequency of occurrence of a set of Caucasian profiles have been estimated with 6 reference databases. The results show that there were greater differences in the frequency estimates when using a database of Afro-Caribbean or Asian profiles, rather than a different Caucasian database. The results further demonstrate the power and robustness of the VNTR DNA profiling technique for forensic casework. PMID- 7495692 TI - Immunocytochemical analysis of hormone mediated nuclear translocation of wild type and mutant glucocorticoid receptors. AB - We have analyzed structural and functional features of the human glucocorticoid receptor (hGR) for their effects on receptor subcellular distribution. COS 1 cells transiently transfected with wild type and mutant hGR cDNAs were assessed immunocytochemically using well-characterized antipeptide antibodies to the hGR. The effect of administration of steroid hormones (and the antiglucocorticoid RU486) on receptor localization was evaluated. Unliganded wild type receptors expressed in COS 1 cells were predominately cytoplasmic. Addition of glucocorticoids or the glucocorticoid receptor antagonist, RU486, resulted in complete translocation of these receptors into the nucleus whereas non glucocorticoid steroids or dibutyryl cAMP were not effective in promoting nuclear translocation. Thus, nuclear translocation was specific for steroids capable of high affinity binding to the hGR. To elucidate the potential role of receptor domains in receptor localization, COS 1 cells transiently transfected with various receptor cDNA mutants were analyzed in a similar manner. Translocation of an hGR deletion mutant lacking the majority of the amino terminus (deletion of amino acids 77-262) was identical to the wild type receptor despite the absence of a transactivation domain. Receptors in which the DNA binding domain was either partially or totally deleted showed an impaired capacity to undergo hormone inducible nuclear translocation. Deletion of the hinge region of the hGR (which also contains part of the nuclear localization signal, NL1) resulted in receptor localization in the cytoplasm. Mutants in the ligand binding domain exhibited two localization phenotypes, exclusively nuclear or cytoplasmic. Receptor mutants truncated after amino acid 550 were found in the nucleus in the presence and absence of hormone consistent with the existence of nuclear localization inhibitory sequences in the ligand binding domain of the receptor. However, a linker insertion mutant (at amino acid 582) which results in a receptor deficient in ligand binding did not undergo nuclear translocation indicating that nuclear localization inhibitory sequences were intact in this mutant. The role of receptor phosphorylation on hormone induced nuclear translocation was also examined. Mouse glucocorticoid receptors which contained mutations of certain hormone inducible phosphorylation sites exhibited translocation properties similar to wild type mGR indicating that these phosphorylation sites on the receptor do not play a major role in hormone inducible nuclear translocation. PMID- 7495693 TI - Hepatic expression of sex hormone-binding globulin associated with the postnatal surge of serum androgen-binding activity in the Djungarian hamster. AB - Serum androgen-binding capacity in Djungarian hamsters, as in many other mammals, increases within days after birth and remains elevated until puberty. This increased activity has been attributed to a hepatic glycoprotein, sex hormone binding globulin (SHBG), but expression of SHBG by the postnatal liver has not been demonstrated. Therefore, a full-length SHBG cDNA was cloned from the liver of neonatal hamsters and the expression of SHBG during development was examined. Hepatic SHBG RNA levels, as measured by both competitive RT-PCR and Northern analysis, were very low in fetal animals but increased significantly within 24 h of birth. Maximal values were maintained for 1 week after parturition, and then declined to basal adult levels. The developmental pattern in hepatic SHBG immunoactivity, as determined by Western analysis, mirrored that of hepatic SHBG mRNA. However, changes in serum SHBG immunoactivity and steroid-binding activity occurred approximately 1 week later. There were no sex differences in the levels of hepatic SHBG mRNA or protein during development, but serum immunoactivity tended to be higher in females at puberty. Sex- and age-related differences in the relative abundance of SHBG isoforms were also noted. Results of these studies demonstrate that Djungarian hamsters express an authentic SHBG and indicate that the postnatal surge in serum androgen-binding activity is due to perinatal up regulation of SHBG expression. PMID- 7495694 TI - Identification of a splice variant of the rat and human mineralocorticoid receptor genes. AB - The sequence of a splice variant of the rat mineralocorticoid receptor (MR) gene is presented. A cDNA clone corresponding to rat MR was isolated from a rat brain cDNA library. Sequence analysis of the region corresponding to the DNA binding domain revealed the presence of a 12 base pair (bp) insertion. Analysis of mRNA from several rat tissues suggests that the variant is less abundant than the wild type in most tissues. The insertion variant is also a product of the human MR gene, the identical splice variant was also observed in human white blood cell mRNA. Unlike other splice variants reported for the MR, this variant alters the encoded protein by the addition of four amino acid residues in the DNA binding domain. The altered protein may influence the affinity of the MR for mineralocorticoid or glucocorticoid response elements. PMID- 7495695 TI - Phosphorylation of the human estrogen receptor by mitogen-activated protein kinase and casein kinase II: consequence on DNA binding. AB - We determined the amino acid and radiolabel sequences of tryptic [32P]phosphopeptides of the purified human estrogen receptor (hER) from MCF-7 cells and Sf9 cells. Serine 118 was identified as a site that was phosphorylated independently of estradiol-binding in MCF-7 cells. Proline is on the carboxy terminus of serine 118, which suggests that the serine-proline may be a consensus phosphorylation site motif for either the mitogen-activated protein (MAP) kinase or p34cdc2 kinase. MAP kinase selectively phosphorylated the recombinant hER in vitro on serine 118 independent of estradiol-binding, whereas p34cdc2 did not phosphorylate the hER. We demonstrated previously that serine 167 of the hER was phosphorylated in an estradiol-dependent manner. We therefore compared the consequence of hER phosphorylation at serine 118 by MAP kinase and phosphorylation at serine 167 by casein kinase II on the receptor's affinity for specific DNA binding. The binding of the hER to an estrogen response element was not altered by phosphorylation with MAP kinase at serine 118 but was significantly increased when phosphorylated at serine 167 by casein kinase II. These data suggest that phosphorylation of the hER by MAP kinase(s) pathways may influence receptor action by a mechanism other than the estradiol-dependent phosphorylation of hER by casein kinase II. PMID- 7495698 TI - Influence of droloxifene (3-hydroxytamoxifen), 40 mg daily, on plasma gonadotrophins, sex hormone binding globulin and estrogen levels in postmenopausal breast cancer patients. AB - Droloxifene (3-hydroxytamoxifen) is a novel antiestrogen currently undergoing clinical investigations for treatment of breast cancer patients. We measured plasma levels of sex hormone binding globulin (SHBG) and the gonadotrophins (LH and FSH) at baseline and after 3 months on treatment in a group of fourteen postmenopausal women treated with droloxifene 40 mg daily. Plasma levels of estrone (E1), estradiol (E2) and estrone sulphate (E1S) were measured in a subgroup of eight patients. Plasma SHBG increased during treatment with droloxifene by a mean value of 16.6% (P < 0.05), while plasma levels of LH and FSH decreased by a mean value of 15.7% (n.s.) and 18.1% (P < 0.05), respectively. Plasma levels of E2 and E1 fell slightly (mean decrease 19.4 and 16.7% respectively, n.s.). On the contrary, plasma levels of E1S increased by a mean value of 23.5% (P = 0.068). The ratio of E1S to E1 and E1S to E2 increased by a mean value of 48.3% (P < 0.025) and 53.2% (P < 0.025), respectively. The effect of droloxifene 40 mg daily on plasma levels of SHBG resembles what is seen during treatment with tamoxifen but occurs to a smaller extent. Contrary to tamoxifen, droloxifene caused a minor suppression of plasma LH levels, suggesting droloxifene to have less estrogen agonistic effects on the pituitary. PMID- 7495697 TI - Glucocorticoid actions on synaptic plasma membranes: modulation of dihydropyridine-sensitive calcium channels. AB - We have previously shown that glucocorticoids accelerate depolarization-induced 45Ca2+ influx in synaptosomes isolated from rat cerebral cortex, indicating that the steroids may modulate voltage-dependent Ca2+ channels. The present study was undertaken to characterize the biochemical action of glucocorticoids on dihydropyridine-sensitive voltage-dependent Ca2+ channels known to be present in brain synaptosomes. The [3H]dihydropyridine labeled site was used as a marker to determine the levels of functional Ca2+ channels. No effect on equilibrium binding of [3H]PN 200-110 was found when membranes from disrupted synaptosomes were incubated with corticosterone as high as 1 microM. However, when intact synaptosomes were first incubated with corticosterone at 37 degrees C and then disrupted, a significant increase in [3H]PN 200-110 binding was found. Steroid incubation of synaptosomes at 0 degree C was ineffective. It appears that metabolic processes requiring intracellular factors were involved in the steroid action. In examining this possibility, [3H]PN 200-110 binding was activated in disrupted membranes by MgATP and Ca(2+)-calmodulin, and corticosterone was found to enhance the activation in a concentration-dependent manner. [3H]PN 200-110 binding to membranes was also activated by incubation with MgATP and cAMP dependent protein kinase, but this activation was not enhanced by the steroid. These findings are consistent with the interpretation that the steroid promotes Ca2+ channel activity by enhancing calmodulin-dependent activation of the channels. The action on voltage-dependent Ca2+ channels in synaptic terminals may well be a mechanism whereby glucocorticoids modulate neuronal activity. PMID- 7495696 TI - A putative step in steroid hormone action involves insertion of steroid ligands into DNA facilitated by receptor proteins. AB - The hypothesis is advanced that hormonal ligands in the steroid/thyroid superfamily act through insertion between base pairs in partially unwound DNA. Using published X-ray coordinates of the complex of the glucocorticoid hormone response element (GRE) with the glucocorticoid receptor DNA binding domain, the interface between the protein and the gene was examined. The site 5'-TG-3'-5'-CA 3' previously shown to accommodate cortisol was found in the first two bases of the GRE half sites, 5'-TGTTCT-3'. These base pairs were sufficiently exposed at the receptor-gene interface to permit access by the steroid. Docking of cortisol into the receptor/DNA complex resulted in a favorable van der Waals energy. Given the general lack of correlation of receptor binding with hormonal activity, we propose that hormone action involves an additional step in which the receptor protein in concert with other transcription factors inserts the hormone into the DNA. This notion provides an explanation for earlier paradoxical observations including structural analogies between base pairs and steroid hormones. The insertion hypothesis suggests that receptor bound ligand facilitates DNA unwinding, stereospecific control of donor/acceptor functional groups on the DNA followed by insertion and release of the ligand between base pairs at 5'-TG-3'-5' CA-3'. PMID- 7495699 TI - Immunocytochemical detection of androgen receptor in human temporal cortex characterization and application of polyclonal androgen receptor antibodies in frozen and paraffin-embedded tissues. AB - Immunocytochemical and biochemical studies have demonstrated the presence of androgen receptor protein in various regions of the rodent and non-human primate cortex. Localization of androgen receptor in the human brain has, however, not been studied as extensively, because of difficulties in obtaining suitable tissue samples. In the present study, we have localized androgen receptors in both frozen and paraffin-embedded temporal cortex from epileptic patients undergoing resection. Polyclonal antibodies were raised against fusion proteins containing fragments of the human androgen receptor protein. The antibodies were affinity purified against the corresponding fusion protein. Immunoprecipitation and Western blotting using extracts from human cell lines demonstrated the specificity of the antibodies for the human androgen receptor and lack of cross reactivity with other steroid hormone receptors. Immunocytochemistry was performed on frozen and paraffin sections of human temporal cortex and in paraffin-embedded benign hyperplastic prostates (BPH), as well as prostate and breast carcinomas, by the streptavidin-biotin-peroxidase method. Antigen retrieval was performed in paraffin-embedded sections using microwave irradiation. Specific nuclear and cytoplasmic immunoreactivity for androgen receptor was detected in neurons, astrocytes, oligodendrocytes, and microglia cells of the temporal cortex. In contrast, only nuclear staining was observed in BPH, prostate and breast carcinomas. Immunoprecipitation of human temporal cortex lysate and subsequent Western blot analysis demonstrated the expression of a 98 kDa immunoreactive protein, slightly smaller than the reported molecular weight of the wild-type androgen receptor. These results provide further evidence for the expression of androgen receptor in the human temporal cortex. The use of these immunocytochemical techniques should enable the retrospective determination of possible changes in androgen receptor expression in a variety of archival paraffin-embedded tissues, including samples of the human central nervous system. PMID- 7495700 TI - Pretreatment with 1,25(OH)2 vitamin D or 24,25(OH)2 vitamin D increases synergistically responsiveness to sex steroids in skeletal-derived cells. AB - We demonstrated previously that vitamin D metabolites modulate the response of bone and cartilage cells to 17 beta-estradiol (E2) and dihydrotestosterone (DHT) both in cell cultures and in vivo. In the present study, we investigated to what extent pretreatment with 1,25(OH)2D3 or 24,25(OH)2D3 would reduce the minimal effective dose of E2, DHT or progesterone (P) required for stimulation of DNA synthesis and creatine kinase specific activity in cultured osteoblast-like ROS 17/2.8 cells and in rat embryo epiphyseal cartilage cells, and to what extent such pretreatment would increase the maximal response. We measured responses to sex steroids after pretreatment of the cells for 5 days with 0.02% ethanol vehicle or with the vitamin D metabolites 1,25(OH)2D3 (0.12 nM), or 24,25(OH)2D3 (1.2 nM) singly or in combination. Pretreatment of ROS 17/2.8 cells with 1,25(OH)2D3, but not 24,25(OH)2D3, increased synergistically their response to E2 but not to P, and did not affect their lack of response to DHT. Pretreatment of epiphyseal cartilage cells with either 1,25(OH)2D3 or 24,25(OH)2D3 increased synergistically their DNA synthetic response to all three steroids, but their CK response only to E2 or DHT. The minimal dose for causing a significant response to E2 in ROS 17/2.8 cells or to either E2 or DHT in epiphyseal cartilage cells was reduced 10-fold after pretreatment with vitamin D metabolites. After pretreatment, the maximal response was more than doubled in ROS 17/2.8 cells; epiphyseal cartilage cells showed a similar 10-fold decrease in the dose required for maximal response to E2 or DHT; the improvement in the response to P was significant only for DNA synthesis. We conclude that pretreatment with the appropriate vitamin D metabolite(s) both reduces by an order of magnitude, or more, the amount of sex steroids needed to stimulate skeletal derived cells and increases synergistically the maximal response of the cells. PMID- 7495702 TI - Pharmacokinetics of tamoxifen in premenopausal and postmenopausal women with breast cancer. AB - We measured the serum levels of tamoxifen and 5 of its metabolites in serum from 7 premenopausal and 9 postmenopausal women during the first 56 days of treatment. The serum levels of N-desdimethyltamoxifen were higher in postmenopausal women compared to premenopausal women (P < 0.02). A similar trend was observed for N desmethyltamoxifen (P = 0.06). PMID- 7495701 TI - Potential mechanisms underlying autoregulation of glucocorticoid receptor mRNA levels in the DHD/K12/PROb rat colonic adenocarcinoma cell line. AB - The DHD/K12/PROb rat colonic epithelial cell line, which was originally derived from a chemically induced adenocarcinoma, expresses functional glucocorticoid receptors (GR) and has been reported to be growth inhibited by glucocorticoid agonists. In the present study the potential mechanisms underlying corticosteroid mediated autoregulation of GR mRNA levels in this colonic cell line were investigated. The GR mRNA levels in the various treatment groups were quantitated via the ribonuclease protection assay using a specific 32P-cRNA probe. Time course experiments demonstrated that in contrast to several other cell lines that are also growth inhibited by glucocorticoids, treatment of confluent monolayers of PROb cells with the pure GR agonist RU 28362 (1 microM) elicits a rapid and significant (65%) down-regulation of GR mRNA levels that is sustained for at least 36 h. This down-regulation, which is also elicited to a lesser extent by weaker GR agonists including corticosterone and aldosterone, is blocked by the GR antagonist RU 38486. The protein synthesis inhibitor cycloheximide was utilized to demonstrate that the initial phase (6 h) of agonist-mediated down-regulation occurs independently of ongoing protein synthesis, although new protein synthesis, perhaps of the GR protein itself, is required to maintain this down regulation. Although agonist-mediated down-regulation in these cells probably occurs primarily at the level of GR gene transcription, inhibition of ongoing RNA synthesis with actinomycin D or 5,6-dichloro-1-beta-D-ribofuranosylbenzimidazole (DRB) demonstrated that during the initial phase (1 h) of this down-regulation, but not following maximal (18 h) down-regulation, RU 28362 treatment also significantly reduces the stability of the GR mRNA. PMID- 7495703 TI - Cloning, sequencing and expression of Pseudomonas testosteroni gene encoding 3 alpha-hydroxysteroid dehydrogenase. AB - We describe the cloning, sequencing and expression of the 3 alpha-hydroxysteroid dehydrogenase (3 alpha-HSD) gene of Pseudomonas testosteroni. A genomic library of P. testosteroni total DNA constructed from SauIIIA digests ligated to an lambda gt11 vector was probed with a polyclonal antibody raised against purified enzyme. Subclones derived from a recombinant phage containing a 1746 bp insert were sequenced and found to contain an open reading frame of 696 bp that corresponds to a protein of 231 amino acid residues. A search for homologous proteins was performed. No similarity was observed when comparing 3 alpha-HSD with known members of the short-chain dehydrogenase family. However a small proteic fragment (80 amino acids) shows homology with the N-terminal sequence of bacterial L7/L12 ribosomal proteins. PMID- 7495704 TI - Nomegestrol acetate, a clinically useful 19-norprogesterone derivative which lacks estrogenic activity. AB - The estrogenic activity of various 19-norprogestin derivatives has been identified by several laboratories. We have previously hypothesized that the estrogenic activity of these compounds stems from the absence of a methyl group at the 19 position, as various progestins that have a methyl group at this position are not estrogens. To test this hypothesis more directly, we now compare the progestin megestrol acetate against its 19-nor analogue nomegestrol acetate. We also compare these compounds to known estrogens (estradiol, norgestrel, RU486) as well as compounds known to be devoid of estrogenic activity at concentrations as high as 10(-6) M (medroxyprogesterone acetate, R5020, ICI 182780). In growth assays using the MCF-7 and T47D:A18 human breast cancer cell lines, we find that only estradiol, norgestrel and RU486 stimulate proliferation, and this effect can be blocked by the pure antiestrogen ICI 182780. Furthermore, in transient transfection studies using a luciferase reporter construct containing three tandem copies of the Xenopus vitellogenin A2 estrogen response element, estradiol, norgestrel and RU486 can stimulate transcription, while none of the other compounds act as estrogens. Transcriptional stimulation by the estrogenic compounds can be blocked by ICI 182780. Our results demonstrate that the lack of a 19-methyl is not the major determinant for estrogenic activity in 19 norprogestins. We suggest that the 17-hydroxyl group more accurately defines estrogenic action. PMID- 7495705 TI - Identification and tissue distribution of a novel variant of 11 beta hydroxysteroid dehydrogenase 1 transcript. AB - A novel variant of 11 beta-hydroxysteroid dehydrogenase 1 (11 beta-HSD1) mRNA was identified from the ovine liver by reverse transcription-polymerase chain reaction (RT/PCR), and was named 11 beta-HSD1C mRNA. Sequence analysis of the RT PCR product revealed that 11 beta-HSD1C mRNA was the product of an alternative exon-splicing within the 11 beta-HSD1 gene in which exon 5 was spliced out. Although it caused a deletion of 48 amino acids in the deduced 11 beta-HSD1 protein, this alternative splicing did not result in a shift within the predicted open reading frame of 11 beta-HSD1 cDNA. Thus, 11 beta-HSD1C mRNA was predicted to code for a protein of 244 amino acids. Using RT-PCR, we also examined the expression of 11 beta-HSD1C mRNA in ovine fetal organs and in maternal myometrium, endometrium, chorion, amnion and placenta. The 11 beta-HSD1C mRNA was expressed ubiquitously, similar to 11 beta-HSD1A mRNA, but at a lower abundance. Furthermore, since levels of 11 beta-HSD1C mRNA were directly related to those of 11 beta-HSD1A mRNA, there is no tissue-specificity for this shorter transcript and the only factor regulating its production appears to be 11 beta-HSD1A mRNA itself. To determine whether 11 beta-HSD1C mRNA encoded a functional enzyme, we inserted the cDNA into the expression vector pRc/CMV, and transfected the construct into Chinese hamster ovary cells. The transfected cells expressed a mRNA of expected size but contained no detectable 11 beta-HSD activity. When combined with cellular extracts of 11 beta-HSD1A cDNA transfected cells, they also did not alter either the dehydrogenase or reductase activity. The functional significance of the 11 beta-HSD1 transcript lacking exon 5 (11 beta-HSD1C mRNA) remains to be determined. PMID- 7495706 TI - Induction of hepatic estrogen sulfotransferase expression by hypophysectomy in female rats. AB - We have examined the effects of hypophysectomy and treatment with thyroxine (T4) on enzyme activity and expression (as determined by immunoblot analysis) of members of the three principal sulfotransferase (ST) sub-families (phenol STs, PST; estrogen STs, EST; hydroxysteroid STs, HST) in cytosols prepared from female Wistar rat livers. The results demonstrate that in female rat liver cytosol, EST activity was decreased by treatment with T4, increased following hypophysectomy and that treatment of hypophysectomized animals with T4 also greatly reduced EST activity. T4 had no significant effect on PST or HST activity in normal animals, but it decreased HST activity in hypophysectomized rat liver cytosol. Immunoblot analysis of these cytosols with antibodies recognising HST and PST indicated that where changes in enzyme activity occurred they mirrored changes in enzyme protein expression. In normal adult female rat livers, EST protein is not expressed, and the small residual activity results predominantly from the action of HST. Hypophysectomy induced EST activity and the expression of EST enzyme protein in female rat liver cytosol, and T4 treatment of hypophysectomized animals reduced the activity to below normal levels without reducing the corresponding enzyme protein levels, indicating that T4 regulation of EST in females is via a post translational mechanism. PMID- 7495708 TI - Metabolism of equilin sulfate in the dog. AB - The metabolism of equilin sulfate was determined in female dogs receiving 2.5 mg/kg of [3H]equilin sulfate alone or in a preparation that contained all the components that are present in the conjugated equine estrogen product Premarin. The pharmacokinetic parameters of total radioactivity indicated that the drug is rapidly absorbed and it has a moderate half-life in plasma. The total radioactivity in plasma following administration of [3H]equilin sulfate as part of a mixture of conjugated equine estrogens had significantly lower peak concentration (Cmax), a lower area under the curve (AUC), a longer terminal half life (t1/2) and a longer mean residence time (MRT) than when [3H]equilin sulfate was given alone, indicating that the other components in the conjugated equine estrogen preparation altered the pharmacokinetics of equilin sulfate. An average of 26.7 +/- 4.4% of the administered radioactive dose was excreted in urine of dogs receiving [3H]equilin sulfate. Again, a significantly lower percentage (21.4 +/- 6.3%, P = 0.023) was eliminated in urine of dogs receiving [3H]equilin sulfate in the conjugated equine estrogen preparation, indicating that the absorption of equilin sulfate was perhaps altered by the other components in the conjugated equine estrogen preparation. Metabolite profiles of plasma and urine were similar. Equilin, equilenin, 17 beta-dihydroequilenin, 17 beta dihydroequilin, 17 alpha-dihydroequilenin and 17 alpha-dihydroequilin were present in both matrices. 17 beta-Dihydroequilin and equilin were the two major chromatographic peaks in plasma samples. 17 beta-Dihydroequilenin and 17 beta dihydroequilin were the major metabolites in urine. In conclusion, following oral administration of [3H]equilin sulfate to dogs, the radioactivity is rapidly absorbed. The disposition of equilin sulfate is altered by the other components that are present in the conjugated equine estrogen preparation Premarin. The reduction of the 17-keto group and aromatization of ring-B are the major metabolic pathways of equilin in the dog. PMID- 7495707 TI - Reptilian sex steroid receptors: amplification, sequence and expression analysis. AB - Sex steroid hormones secreted by the gonads play a central role in the reproduction of all vertebrates. In addition to direct effects on gametogenesis, sex steroid hormones are important in sexual development, brain organization, and sexual behavior. The actions of sex steroid hormones are mediated primarily by ligand-dependent transcription factors, or receptors which bind to specific sequences of the DNA and alter the transcription rates of nearby genes. We have used the polymerase chain reaction to amplify cDNA fragments of the estrogen receptor, progesterone receptor and androgen receptor from the unisexual whiptail lizard, Cnemidophorus uniparens. The lizard steroid hormone receptors share a high degree of sequence homology to the steroid hormone receptors of other vertebrates. Ribonuclease protection assays demonstrate that both estrogen receptor mRNA and progesterone receptor mRNA are increased in the oviduct during vitellogenesis and after estrogen treatment. This report demonstrates the utility of the polymerase chain reaction to generate species specific probes for comparative molecular studies and provides the first report of cDNA sequences for reptilian steroid hormone receptors. PMID- 7495710 TI - White blood cell count in healthy Sierra Leoneans. AB - The lack of standard local values for use as reference values has been a problem facing our haematological units in Sierra Leone for a long time. As a preliminary step towards the establishment of standard local values, one hundred and sixty apparently healthy Sierra Leoneans between the ages of 10 years to 50 years and above living in Freetown were investigated for the determination of total and differential white blood cells counts. The total white cell count was done by using the neubeur counting chamber whilst the differential white cell count was done from a thin blood smear stained with leishman's stain. A mean white blood cell count of 5.0 x 10(9)/L was obtained for the study group with a range of 2.4 12.0 x 10(9)/L. The study did not show any significant difference in white blood cell count between males and females. A mean differential neutrophil count of 56% was obtained for the study group and a mean differential lymphocyte count of 37% were obtained for the study group. The leucopenia obtained for the study group is thought to be due to the neutropenia and is probably thought to be a genetically determined characteristic of the indigenous Africans. PMID- 7495709 TI - Childhood deaths from anaemia in Accra, Ghana. AB - Severe anaemia has remained a major cause of morbidity and mortality in children of Southern Ghana since the early 1960s. 71.1% of 15450 children attending the Korle Bu Teaching Hospital, Accra referred to the laboratory for haematological studies had haemoglobin (HB) levels below 11.0 Gm/dl while 27.7% of anaemic patients had Hb levels below 7.0 gm/dl. Indeed, 71.1% of children with severe anaemia had Hb levels below 5.0 gm/dl, thus requiring urgent blood transfusion. Though the Department of Child Health alone utilised 32.2% of total blood processed by the National Blood Transfusion Service at Korle Bu, as many as 259 (58.1%) of the 554 deaths in the emergency rooms per annum in children beyond the neonatal period were related to severe anaemia. Iron deficiency was the commonest cause of anaemia and contributed further to the anaemias of sickle cell disease and protein--energy malnutrition. In the light of the significant decline in the prevalence of childhood anaemia in the developed world following improved counseling in nutrition, fortification of foods with iron, and iron supplementation to infants and school children, and the documented attendant improvement in growth velocity and intellectual performance we support the planned national anaemia survey and recommend for early consideration iron supplementation to older infants and pre-school children at risk. PMID- 7495711 TI - Sudden deaths: cardiac and non-cardiac in children in Accra. AB - Sixteen (16) paediatric patients have suffered sudden deaths in Accra, Ghana over the last 15 years from 1980 to 1994. There were eight (8) females and 8 (eight) males involved; the M:F ratio is 1:1. Eight (8) of them were known heart disease patients; six (6) were not known to have any chronic illness, one had sickle cell anaemia and the other had abdominal tuberculosis. Cardiac: non-cardiac ratio is 1:1 in this report. The overall frequency of sudden deaths among children this author alone has been associated with, in Accra to date is about one per year. PMID- 7495712 TI - Seroprevalence of HIV infection among patients with pulmonary tuberculosis in Yaounde-Cameroun. AB - To determine the seroprevalence of infection with the human immunodeficiency virus (HIV) in pulmonary tuberculosis patients, we conducted an HIV serosurvey in 162 inpatients with pulmonary tuberculosis, in Yaounde Cameroon. HIV seroprevalence in hospitalized pulmonary tuberculosis patients was found to be 9.9% as compared to the seroprevalence rate of 2.2% for Yaounde, the area from which the patients we studied came. No differences were observed in the clinical, radiological and bacteriological features between HIV seropositive and seronegative patients with pulmonary tuberculosis. PMID- 7495713 TI - Birth weights of Nigerian newborn infants--a review. AB - Birthweight (BW) is an important denominator for a continuing re-appraisal of a Nation's health care delivery and neonatal viability. A constant surveillance of BW is therefore of utmost importance and a decline, when detected should allow for selecting areas at great risk, identifying causes for the decline and proffering possible corrections. The mean BWs of Nigerian babies differ within the country and, in general, are lower than those of caucasian counterparts. Information is accruing that low BWs are created by factors most of which can be prevented or significantly modified. Mean BWs in Nigeria are highest in the South (lagos: 3380g) and the Middle-Belt (Jos: 3280g: male, 3186g: female). The goal of this synopsis is to summarize and critically analyse the information available to us and to show the lacunae in our knowledge where future research is imperative. PMID- 7495714 TI - Diabetes mellitus induced by thecoma-fibroma tumour of the ovary: case report. AB - A case of Thecoma-fibroma secreting oestrogen to cause insulin intolerance in a post-menopausal woman is presented. Blood levels of sugar became normal 24 hours after removal of the tumor. The possible mechanism is presented. PMID- 7495715 TI - Problems in extubation of endotracheal tube in anaesthesia in the West African sub-region. AB - Three Retrospective Case Studies of the problems often faced with extubation of endotracheal tube during routine endotracheal anaesthesia is reported to alert anaesthetists about endotracheal tubes with intact and inflatable cuffs which could pose potential dangers to safety of patients in Anaesthetic practice, especially in the West-African Sub-region because of our practice of re-using both re-usable and disposable endotracheal tubes. PMID- 7495716 TI - Prevention--how much of it in our health care? PMID- 7495717 TI - Trial of Badoe's maintenance solution. AB - The adult West African on intravenous fluid therapy requires 130 mmol of sodium, 50 mmol of potassium and 3L of water daily. At least 100g of glucose is usually provided. Three liters of Badoe's Maintenance Solution which contains 43.3 mmol of sodium of sodium, 16 mmol of potassium, 0.65 mmol of calcium, 9 mmol of bicarbonate, 51.7 mmol of chloride and 100g of sorbitol per litre, provides all the daily requirements of sodium, potassium and water. It also provides about 5000j of energy, nearly two-thirds of the resting energy expenditure. The trial in seven ill patients from the first day after operation has shown that the serum concentration of sodium and potassium and the urinary output of patients on 3L of Badoe's Maintenance Solution alone daily remain normal. Badoe's Maintenance solution is, therefore, recommended as the standard maintenance solution for patients in balance who need intravenous fluid therapy. It simplifies fluid therapy for medical and nursing staff as only one solution is used. PMID- 7495718 TI - Prevalence of bacterial otitis media in primary school children in Enugu Surburb, Enugu state, Nigeria. AB - Out of a total school population of 3,070 pupils examined for discharging ear due to bacterial etiology, 44 (1.4%) had discharging ear. Forty eight species of bacteria belonging to 5 genera were isolated. These included, Staphylococcus aureus, 16 isolates; Pseudomonas aeruginosa, 10; escherichia coli, 11; Proteus species, 7; Bacillus sp. 4. The highest prevalence of infection (3.0%) was recorded in central school Akwuke, Awkunanaw. Analysis of the data showed that infection rate was highest among the 6-9 year old age group (61%) and lowest among the 15 years and above age group (5%). Boys recorded a higher prevalence of infection (55%) than girls (45%). However the difference was not statistically significant (x = 2.08, p > 0.01). All the organisms isolated were sensitive to genticin but resistant to other commonly used antibiotics. PMID- 7495719 TI - Traditional methods of treatment of cataract seen at Korle-Bu Teaching Hospital. AB - Fourteen eyes in eleven patients treated by the traditional methods for cataract were evaluated. Three different methods of treatment were classified from the cases: intracapsular (the traditional couching), extracapsular, and zonulysis. The operations were carried out on the left eye in 64%, bilateral in 27%, and right in 9%. Complications noted were uveitis, glaucoma, lens in pupil, posterior capsule opacification, cystoid macular oedema (CMO) but infection was conspicuously absent. Some of the results were good. With those that did not appear acceptable and could be further improved patients either received partial management or did not bother. Pain was the worrying symptom. These results of the traditional methods are not desirable but until intra-ocular lens implantation is widely practised in Ghana traditional methods are likely to compete with orthodox methods to a variable degree. PMID- 7495720 TI - Disputed paternity: experience in Ile-Ife, Nigeria. AB - Results of paternity tests carried out in our hospital between September, 1984 and December, 1993 were analysed. There were 44 cases consisting of 37 (84.1%) groups of four each: the child, the mother and the two alleged fathers, and 7 (15.9%) groups of three, including the disputed child, the mother and the alleged father. Most of the disputed victims were children under age 10 years (97.7%), the oldest 'child' in dispute was 35 years. The total cumulative paternal exclusion rate with our standard techniques was only 27.5% consisting of 15.9% for ABO blood group system, 2.3% for Rhesus-D system and 9.3% for haemoglobin electrophoresis analysis. This is far below the exclusion rate of up to 97% and 99.9% obtained with the more reliable tests such as HLA and DNA analyses, respectively. There is a need to introduce a more reliable screening package for paternity testing in our specialised hospitals in order to ease the associated medicolegal problems since all the cases but one seen were referred from law courts. PMID- 7495721 TI - Needs assessment in demand reduction programmes with special reference to African countries. AB - Drug abuse trends in African countries indicate an urgent need for intensive demand reduction packages to address the problem. However, enormous obstacles exist as many of these countries lack the necessary resources to assess the basic needs essential to implementing empirically-based and cost-effective programmes. This article reviews general principles of needs assessment and provides some practical guidelines appropriate to the African context. Effective cooperation and collaboration at the local, national and the international level is needed to attain the desired goals of needs assessment in these countries. PMID- 7495722 TI - External intestinal fistulae in Port Harcourt. AB - Twenty cases of established external faecal fistulae treated at the University of Port Harcourt Teaching Hospital (UPTH) over an eight year period were studied to find out the peculiar characteristics of this problem in this part of Nigeria. The male/female ratio was 3:1 Surgical complications constituted the commonest cause of fistula formation (65%). 80% of the patients were referred to the UPTH from peripheral hospitals and clinics. All patients were initially treated conservatively by nutritional build up, correction of fluid and electrolytes, control of infection and, where necessary, blood transfusion. 15% of fistulae closed spontaneously on conservative treatment while 55% required surgery for a successful outcome. The overall mortality was 25%. PMID- 7495723 TI - Role of the OCTA site in regulation of IgH chain gene transcription during B cell activation. AB - We have evaluated the importance of the OCTA site in both the promoter and enhancer regions for the induction of enhancement of IgH chain gene transcription after B cell activation. These studies show that although occupancy of the OCTA site in the promoter is critical for basal transcription of the mu gene, it is not necessary for the increase in transcription induced by in vivo activation. On the other hand, the OCTA site in the enhancer is necessary for neither basal transcription nor in vivo activation of transcription; however, occupancy of this site is required for further up-regulation in transcription of the mu gene in pre activated cells. These results indicate that different mechanisms may be involved in the activation of resting versus in vivo stimulated B lymphocytes. The findings are discussed in relation to the phenotype described for Oct-2-deficient mice. PMID- 7495724 TI - Immunological characterization of antigenic domains on human IL-2 receptor beta subunit: epitope-function relationships. AB - Five mAb directed at the IL-2R beta chain were analyzed for their binding and functional properties. They define three epitopes on a recombinant soluble beta chain or on the beta chain expressed at the surface of YT-2C2 cells. Epitope 1 (A41 and 6E8 mAb) is part of the IL-2 binding domain, whereas epitope 2 (CF1 and 6E10 mAb) is not involved in IL-2 binding. Epitope 3 (6B5 mAb) also partly overlaps the IL-2 binding domain but does not overlap epitopes 1 and 2. None of the mAb can by themselves inhibit IL-2 induced proliferation of a human activated T cell clone. Only epitope 1 mAb can synergize with an anti-alpha chain mAb to inhibit this proliferation. Using epitope 1 and 2 mAb as well as a purified, recombinant form of the IL-2R beta chain extracellular domain, an ELISA-based immunoassay was set up which allows the quantitative determination of soluble and detergent solubilized IL-2R beta chains. Epitopes 1 and 2 are in non-competitive interaction: the binding of a mAb to one epitope decreases the affinity of a mAb for the second epitope. Epitope 2 mAb have binding stoichiometries (approximately 16,000 sites/cell) which are approximately 80% higher than that of epitope 1 mAb and IL-2 itself (approximately 9000 sites/cell). Upon binding of epitope 2 mAb, the stoichiometries of epitope 1 mAb and IL-2 are increased to reach the stoichiometry of epitope 2 mAb.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7495725 TI - Involvement of alpha 1 and alpha 4 integrins in gut mucosal injury of graft versus-host disease. AB - We have investigated the involvement of adhesion molecules in the lymphocyte infiltration associated with acute intestinal graft-versus-host disease (GVHD) induced by injection of C3H lymph node cells into irradiated (C3H x DBA/2)F1 mice. First we analyzed the expression profile of adhesion molecules including alpha 1, alpha 2, alpha 4, alpha 5, alpha 6, alpha L and beta 7 integrins, CD44 and L-selectin of lymphocytes from lymph nodes and gut mucosa in normal mice. In normal mice, intraepithelial lymphocytes (IEL) and lamina propria lymphocytes (LPL) uniquely showed increased expression of alpha 1, alpha 2 and beta 7 integrins, and decreased expression of L-selectin compared with that of lymphocytes of the lymph nodes and Peyer's patches. In mice with GVHD, IEL and LPL of donor lymph node cells origin underwent phenotypic changes characterized by the increased expression of alpha 1, alpha L and beta 7 integrins, and the loss of L-selectin. The expression profile of adhesion molecules on IEL and LPL of GVHD mice resembled that of normal mice except for the lack of alpha 2 integrin. Treatment of GVHD mice with anti-alpha 1, -alpha 4 or -beta 7 integrin antibody alone partially prevented the mucosal pathology of intestinal GVHD, whereas only mice treated with anti-alpha 1 showed reduced donor lymphocytic infiltration into the intestinal mucosa. In contrast, treatment with anti-alpha L or anti-CD44 antibody did not affect the intestinal GVHD. Furthermore, dual blockade of both alpha 1 and alpha 4 integrins completely inhibited the mucosal pathology and donor lymphocyte infiltration of intestinal GVHD.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7495726 TI - The human I alpha 1 region contains a TGF-beta 1 responsive enhancer and a putative recombination hotspot. AB - It appears that the switch recombination machinery of a B lymphocyte targets preferentially unrearranged heavy chain genes that have been rendered transcriptionally active. Transcriptional activation of the 'germline' human C alpha 1 and C alpha 2 genes is triggered by TGF-beta 1 and is controlled by proximal positive and distal negative regulatory elements residing upstream of the alpha 1 and alpha 2 switch regions respectively. In this report we characterize the positive proximal regulatory elements and analyse their interaction with DNA binding proteins. Our data demonstrate that a 100 bp fragment that contains a cAMP responsive element (CRE)/activating transcription factor (ATF) motif, a putative Ets binding site and an element that is created by two previously described neighbouring direct repeats (DRE), can increase the basal level of transcription and confer TGF-beta 1 inducibility to a heterologous promoter in an orientation- and position-independent manner. Ubiquitously expressed DNA binding proteins interact specifically with the CRE/ATF, the Ets site and the DRE element. Additionally, nuclear proteins interact with sequences which are located downstream of this enhancer are not essential for transcription in the transient expression assays utilized; however, they contain motifs that have been previously implicated in regulating DNA recombination events. These motifs include a Chi motif and a Chi-like element previously found in the recombination hotspot region of the Bcl-2 proto-oncogene and close to chromosomal breakpoints in T-ALL lines. Our findings raise the possibility that the intervening region associated regulatory elements in addition to regulating the transcriptional activation of the Ig heavy chain genes could also facilitate the physical interaction of transcription and recombination controlling molecular mechanisms. PMID- 7495727 TI - CD4-independent in vivo priming of murine CTL by optimal MHC class I-restricted peptides derived from intracellular pathogens. AB - CTL combat intracellular pathogens by killing infected cells. The molecular targets of their attack are foreign peptides bound to self MHC encoded class I molecules. Immunization of mice with peptides containing CTL determinants was shown to elicit CD4-dependent CTL. Here, we have achieved in vivo CTL priming with naturally processed 8-10 amino acid long class I-restricted peptides emulsified in an adjuvant. A potent, reproducible and physiologically relevant response was obtained using peptides from an intracellular bacterium and five viruses (including HIV) in two murine MHC haplotypes. This method is suitable for multiple vaccination, since a 'cocktail' of peptides derived from three pathogens elicited effector CTL against each pathogen. Most importantly, peptide-induced CD8+CD4- CTL were CD4(+)-independent. These results have implications for CTL induction in situations where CD4 T cells are depleted or compromised, as is the case in HIV infection. PMID- 7495728 TI - The accumulation of B220+ CD4- CD8- (DN) T cells in C3H-lpr/lpr mice is not accelerated by the stimulation of CD8+ T cells or B220+ DN T cells with staphylococcal enterotoxin B and occurs independently of V beta 8+ T cells. AB - Mice homozygous for lpr or gld develop lymphoproliferative disease characterized by the progressive accumulation of functionally impaired B220+ double-negative (DN) T cells and primed CD4+ and CD8+ T cells. The mechanisms leading to the accumulation of these T cells subsets are poorly understood but are clearly dependent on lack of expression of Fas in lpr mice and expression of defective FasL in gld mice. A role for V beta 8+ T cells also has been reported. Recently, a variety of experimental approaches revealed that the majority of B220+ DN T cells are derived from MHC class I-selected CD8+ precursors. Here we used the potent mitogen, staphylococcal enterotoxin B (SEB): (i) to examine the effects of defective Fas-FasL expression on the deletion of peripheral V beta 8+ T cells in 6- to 8- and 20-week old C3H-lpr and -gld mice, (ii) to determine the immunocompetence of B220+ DN T cells in vivo, and (iii) to determine if activated V beta 8+ CD8+ T cells can differentiate into B220+ DN T cells. The role of V beta 8+ T cells in the accumulation of B220+ DN T cells also was reinvestigated. These studies showed that deletion pathways independent of Fas-FasL expression function in young lpr and gld mice and delete CD4+ T cells more efficiently than CD8+ T cells. As the mice age, these alternative pathways become less effective and this may explain the progressive accumulation of memory T cells. No abnormalities in tolerance induction were observed in young or diseased mice. Stimulation of +/+, lpr and gld V beta 8+ CD8+ T cells induced the expression of B220. B220 levels were maximal 2 days after SEB and were undetectable 5 days later, suggesting that B220 is a transiently expressed activation marker on CD8+ T cells. Neither the B220+ V beta 8+ CD8+ T cells nor other V beta 8+ T cell populations converted with detectable frequency into B220+ DN T cells after single or multiple doses of SEB. B220+ DN T cells, which are functionally anergic in vitro, did not proliferate or undergo deletion after SEB stimulation indicating that these cells also are functionally impaired in vivo. In contrast to previous reports, chronic elimination of V beta 8+ T cells had no effect on the accumulation of B220+ DN T cells.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 7495729 TI - Somatic hypermutation of VHS107 genes is not associated with gene conversion among family members. AB - Murine B cells proliferating in the germinal centers of peripheral lymphoid tissue accumulate mutations in their rearranged variable regions, a diversification process which contributes to affinity maturation of the antibody response. The highly targeted nature of the hypermutation process could be explained by a somatic gene conversion mechanism. Well characterized examples of such an activity in B cells are seen during diversification of the chicken and rabbit Ig repertoires. The genomic organization, low complexity and high degree of homology exhibited by the four members of the murine VHS107 gene family suggested that these gene segments may be suitable candidates for the search of gene conversion events derived from upstream VHS107 counterparts. After an immune response to a complex T cell-dependent antigen (sheep red blood cells), rearranged V13, V11 and V1 genes were isolated from splenic extrafollicular and germinal center B cells. Extensive somatic mutation was evident in V11 and V1 sequences. When these sequences were examined, as well as V1 sequences isolated from phosphorylcholine-specific hybridomas, the observed nucleotide changes were not associated with any gene conversion between family members, suggesting instead that they arose by a mechanism which introduces point mutations. PMID- 7495730 TI - Composition of TCR-CD3 complex in human intestinal intraepithelial lymphocytes: lack of Fc epsilon RI gamma chain. AB - Human intestinal intraepithelial lymphocytes (iIEL) are a unique population of predominantly CD8 alpha beta+, TCR alpha beta+ lymphocytes and, to a lesser extent, TCP gamma delta+ lymphocytes that proliferate poorly to anti-CD3 mitogenic signals but display significant cytolytic activity. Studies in mouse model systems have shown that the gamma chain of the high-affinity receptor for IgE (Fc epsilon RI gamma) may substitute for the zeta chain in the TCR-CD3 complex of iIEL. This has suggested that the functional properties of these cells may be associated with an altered composition of the TCR-CD3 complex. We therefore analyzed the TCR-CD3 complex of normal human iIEL. One- and two dimensional non-reducing/reducing SDS-PAGE analysis of CD3 gamma, CD3 delta, CD3 epsilon, zeta and Fc epsilon RI gamma chain immunoprecipitates of cell surface radiolabeled proteins with subunit-specific antibodies revealed a TCR-CD3 complex without associated Fc epsilon RI gamma chains. Thus, normal human iIEL contain a TCR-CD3 complex that consists predominantly of zeta homodimers in association with the alpha beta TCR and CD3 gamma, delta and epsilon, similar to the majority of peripheral lymphocytes. This indicates that the distinct properties of human iIEL are not associated with substitutions of the Fc epsilon RI gamma chain in the TCR-CD3 complex. PMID- 7495732 TI - Affinity for class II MHC determines the extent to which soluble peptides tolerize autoreactive T cells in naive and primed adult mice--implications for autoimmunity. AB - The N-terminal peptide (Ac1-9) of myelin basic protein (MBP) is the immunodominant encephalitogenic epitope in H-2u mice. Previous studies have defined the role of amino acid residue 4 in binding to I-Au. Accordingly, substitutions at this residue have generated peptides spanning a wide range of affinities for the MHC. In the present study, we have tested the tolerogenicity of three of these peptides. Ac1-9, Ac1-9[4A] and Ac1-9[4Y], by administering these to mice i.p. in the absence of adjuvant. Significantly, mice treated with the high affinity analogues Ac1-9[4A] and Ac1-9[4Y] prior to immunization became less susceptible to Ac1-9-induced experimental autoimmune encephalomyelitis (EAE), whereas those given the low affinity peptide Ac1-9 were only moderately protected. T cell priming, as assessed by in vitro proliferative and lymphokine assays, demonstrated a direct correlation between the level of disease inhibition and T cell unresponsiveness. In treatment studies, Ac1-9 and Ac1-9[4Y] were also shown to be effective when given on the first day of disease onset. Priming of T cells, when measured by proliferation in vitro, however, became more resistant to inactivation when soluble peptides were administered close to the day of assay. Kinetic studies revealed that tolerance could be achieved in primed mice but that this takes time to develop.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7495731 TI - Long-term cultured CD40-activated B lymphocytes differentiate into plasma cells in response to IL-10 but not IL-4. AB - We compared the effects of IL-10 and IL-4 on the functions of B lymphocytes triggered through their CD40. During the initial phase, IL-10 was as potent as IL 4 in inducing the expansion of viable B cells. Then, cellular expansion slowed down and after approximately 3 weeks the number of B cells started to decline. While the combination of IL-10 and IL-4 was synergistic during the first 2 weeks of culture, B cell recovery declined after 3 weeks, indicating that IL-10 prevails over IL-4. Those effects were not restricted to a specific B cell subset as both sIgD+ B cells and sIgD- B cells behaved in a similar way, though the latter population responded with a slightly accelerated kinetic. Inverted microscope examination and scanning electron microscopy showed that in response to IL-10, CD40-activated B cell cultures were heterogeneous with loose aggregates of cells as well as free floating large ovoid cells. In contrast, in the presence of IL-4, CD40-activated B cell cultures were essentially composed of tight cell clumps. IL-10 progressively induced all B cells to differentiate into non replicating cells with intracytoplasmic Ig that secreted Ig at a high rate. Cytologic analysis indicated that IL-10 cultured cells display a basophilic cytoplasm with an arcoplasm and a low nucleus/cytoplasm ratio. Transmission electron microscopy demonstrated that when IL-10 was added to the culture, B cells displayed structures for excretion with extended endoplasmic reticulum and dilated cisternae containing paracrystalline structures, typical of plasmablasts cells. Taken together, these results indicate that IL-10 acts as a plasma cell differentiation factor for CD40-activated B cells. PMID- 7495733 TI - The ontogeny of class-regulation of CD4+ T lymphocyte populations. AB - The differential class-regulation of CD4+ T lymphocyte populations is believed to play a major role in determining the qualitative behaviour of the immune system, and in the fate of immune responses in particular. In this article we propose a model for the dynamics of the Th1 and Th2 subpopulations. We put forward the concept of an 'antigenic niche' which allows us to postulate that the key feature underlying the regulation of Th differentiation pathways is the population dynamics of the lymphocytes themselves. Using this model we are able to account for a number of well established experimental observations which were hitherto apparently unrelated and poorly understood. This suggests that our simplified model might be capturing some essential features of the immune system. PMID- 7495734 TI - Expression of an ovalbumin-specific V beta 8.2 TCR transgene inhibits collagen arthritis in B10.Q mice. AB - Previous studies have illustrated the importance of T cells bearing alpha beta TCRs in the induction and development of collagen induced arthritis (CIA) in mice. However, the scope of TCR usage in CIA has yet to be clearly defined. Given the inherent diversity of the TCR repertoire, the relative flexibility of the arthritogenic TCR repertoire specific for type II collagen (CII) is not clear. Therefore, we chose to examine the influence of a highly skewed TCR repertoire on CIA. Arthritis susceptible B10.Q (H-2q) mice were mated with C57L (H-2b) animals expressing an ovalbumin-specific V beta 8.2 TCR transgene (Tg) and Tg+ offspring were further backcrossed to B10.Q. Homozygous H-2q/q, V beta 8.2 Tg+ mice displayed a high level of V beta 8.2+ T cells in peripheral blood. However, expression of some endogenous V beta TCR, such as V beta 14, was still detected. Upon immunization with bovine CII in adjuvant, V beta 8.2 Tg+ mice were highly resistant to CIA when compared with Tg- littermates. Analysis of sera demonstrated a marked reduction in antibody specific for homologous mouse CII as well as heterologous bovine CII in Tg+ animals. Interestingly, V beta 8.2 Tg+ mice still mounted good antibody responses following immunization with human thyroglobulin, indicating that the skewed TCR repertoire affected anti-CII but not antithyroglobulin responses. Thus, our findings show that constraints placed on the TCR repertoire inhibit pathogenic responses against CII and suggest that in H-2q mice the arthritogenic TCR repertoire bears only limited flexibility. PMID- 7495735 TI - Are CD4+ Th1 cells pro-inflammatory or anti-inflammatory? The ratio of IL-10 to IFN-gamma or IL-2 determines their function. AB - Human CD4+ T cells have, like their murine counterparts, been classified on the basis of their cytokine profile. Th1 cells produce IL-2 and IFN-gamma, but little or no IL-4. Th2 cells produce IL-4 but not IFN-gamma or IL-2, and Th0 produce IL 2, IL-4 and IFN-gamma. As IL-2 is the most potent T cell growth factor and IFN gamma is the strongest activator of macrophages it is not surprising that CD4+ Th1 cells are considered to be pro-inflammatory. However, unlike results in the mouse, where IL-10 is only produced by Th2 cells, human IL-10 is produced by Th0, Th1 and Th2 cells. Hence some human Th1 cells are capable of producing both pro inflammatory (IL-2, IFN-gamma) and anti-inflammatory (IL-10) cytokines, therefore the function of these cells may not be accurately encapsulated by the 'Th1' terminology. We thus investigated the correlation of cytokine production and function in human CD4+ Th1 clones. Cytokine production (IL-2, IFN-gamma, IL-10) was measured in supernatants by ELISA after stimulation with solid-phase anti CD3. The capacity of these supernatants to activate or inhibit T cell proliferation or LPS induced TNF-alpha production by monocytes was assessed. The ratio of IL-2/IL-10 or IFN-gamma/IL-10 was of critical importance in determining the function of the supernatants. The inhibitory effects were verified to be due to IL-10, as they were neutralized by anti-IL-10 mAb.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7495737 TI - Development of retrovirally marked human T progenitor cells into mature thymocytes. AB - Retroviral vectors have been used in most human gene therapy trials that have been undertaken. Many of these therapies have focused on the introduction of genes into hematopoietic stem cells with the goal of obtaining expression in the mature T lymphocytic progeny. It has proven difficult to achieve expression in the lymphoid lineage, although several groups have demonstrated low expression of transduced genes in the myeloid lineage. In this study we used an in vitro thymic organ culture in which stem/progenitor cells can develop into T cells and all intermediate stages can be studied and manipulated to investigate the fate of a retrovirally introduced Escherichia coli LacZ gene in this system. Here we show that certain conditions can transduce Jurkat T cells, three different antigen specific T cell clones and CD34+CD3-CD4-CD8- thymocytes (progenitor T cells) with high (> 80%) efficiency. Moreover, retroviral transduction with the LacZ gene does not inhibit T and NK cell differentiation of progenitor cells in fetal thymic organ cultures (FTOC). The LacZ gene also is functionally expressed at all stages of development, although the expression decreases somewhat during differentiation. This experimental system, combining FTOC and retroviral transduction, provides a genetic tool for the study of human T cell development. PMID- 7495736 TI - The two novel MHC class II transactivators RFX5 and CIITA both control expression of HLA-DM genes. AB - MHC-encoded HLA-DMA and -DMB molecules are atypical MHC chains that play an essential role in antigen presentation by MHC class II molecules. They resemble both MHC class I and II molecules but are not expressed at the cell surface. From the study of MHC class II regulatory mutants, it was found recently that two novel transactivators, CIITA and RFX5, are essential for the control of MHC class II gene expression. We report here that CIITA and RFX5, although operating at different levels of transcriptional control, are also both essential regulators of HLA-DMA and -DMB genes. This is true for both the constitutive and the inducible mode of DM gene expression. Indeed, both CIITA and RFX5 cDNA can correct the HLA-DMA and -DMB gene expression defect in the respective regulatory mutants. The involvement of these two transcription factors accounts for the coordinate expression of MHC class II and HLA-DM, two sets of molecules that perform quite different functions in the overall process of antigen presentation. PMID- 7495738 TI - Selection of TCR V alpha by MHC class II predicts superantigen reactivity. AB - Recognition of superantigens by T cells predominantly involves the TCR V beta region. The contribution of reactivity from the non-V beta portion of the TCR remains less clear. We have investigated the V alpha repertoire of T cells bearing one V beta element responding to different superantigen or polyclonal stimuli. The data indicate that the V alpha chain is not directly involved in superantigen recognition, since unrelated superantigens do not stimulate cells bearing different V alpha elements. Instead, analysis of V alpha elements used preferentially in the CD4+ or CD8+ cells, shows that the V alpha-regions predominating in the superantigen response are the same V alphas that are positively selected into the CD4+ lineage. Thus the ability of a V alpha-region to either aid or hinder the interaction with class II predicts its frequency in a superantigen-responding population of T cells. PMID- 7495739 TI - Characterization of the epitope recognized by a mAb that reacts differentially with murine suppressor T cells. AB - Although reliable antibodies are available that distinguish human suppressor T (Ts) cells from CTL and other T cells, few are available for murine Ts cells. We have developed a mAb (984D4.6.5) that, in the presence of complement, depletes alloantigen-specific Ts cells but not CTL. This antibody recognizes activated Ts cells but not their precursors. In these studies, flow cytometric analysis demonstrates that 984D4.6.5 reacts with several Ts cell hybridomas, cloned Ts cell lines and WEHI-3 (a myelomonocytic tumor cell line). Reactivity was not detected with BW5147, Th cell hybridomas, cloned Th cells, CTL lines and hybridomas, B cell lines, thymocytes, splenocytes, bone marrow cells nor a variety of tumor cells. Among 984D4.6.5 positive lines, expression is heterogeneous and the number of cells expressing high levels of the epitope is increased when the hybridomas are maintained at a relatively high cell density. Neuriminidase and pronase deplete the epitope recognized by mAb 984D4.6.5. Protein synthesis and glycosylation inhibitors also reduce expression of this epitope. These observations suggest that the epitope recognized by 984D4.6.5 is a carbohydrate linked to a polypeptide. This antibody was tested by ELISA for binding to a large panel of carbohydrates and glycolipids coupled to BSA. The only one that bound 984D4.6.5 was LS tetrasaccharide c (NeuNAc alpha 2-6Gal beta 1-4GlcNAc beta 1-3Gal beta 1-4Glc), an O-linked carbohydrate. Comparative analysis shows that both the sequence and the linkage of these sugars are essential to the reactivity with the 984D4.6.5 antibody. This epitope is expressed by a glycoprotein of approximately 200 kDa, as shown by Western blots. The identity of this glycoprotein remains to be determined, but indirect evidence suggests that it is not CD45. PMID- 7495740 TI - CD86 (B70/B7-2) on endothelial cells co-stimulates allogeneic CD4+ T cells. AB - In vascularized organ transplantation, vascular endothelial cells (EC) confronting recipient T cells are potentially significant APC initiating cellular immune responses that lead to rejection. In the present study, we studied the ability of human EC to stimulate allogeneic T cells and the co-stimulatory molecules involved in this response. On both human umbilical vein endothelial cells (HUVEC) and microvascular endothelial cells (MVEC), MHC class I, intercellular adhesion molecule (ICAM)-1 and CD86 were constitutively expressed as assessed by flow cytometry. After IFN-gamma treatment, MHC class II expression was induced, and MHC class I and ICAM-1 were up-regulated. In contrast, the expression of CD86 was unchanged and CD80 was undetectable even after IFN-gamma treatment. Highly purified CD4+ T cells proliferated in response to IFN-gamma treated allogeneic HUVEC and MVEC, and this response was efficiently blocked by mAb to MHC class II, ICAM-1 and CD86. Furthermore, the addition of anti-CD86 mAb to the primary culture with allogeneic EC resulted in the induction of alloantigen-specific anergy. These results suggest that CD86 expressed on EC plays a critical role in initiating cellular immune responses to vascularized allografts and would be an important target for immune intervention. PMID- 7495742 TI - Gene-targeted B-deficient mice reveal a critical role for B cells in the CD4 T cell response. AB - The role of B cells in promoting T cell responses is still controversial. In this study, we use JHD mice which have a targeted mutation in the JH gene and are thus rendered deficient in B cells to address this issue. We show here that immunization of JHD mice with soluble antigen fails to prime CD4 T cells, for either clonal expansion or delivery of immunological help for antibody responses. This lack of CD4 T cell priming in JHD mice corresponds to a 3- to 9-fold lower co-stimulatory activity of antigen-presenting cells (APC) from the JHD mice, as measured by anti-CD3-induced proliferative responses of CD4 T cells. This in turn is due to a defect of APC from JHD mice in response to T cell-mediated induction of co-stimulatory activity. As the development of macrophages and dendritic cells is unaffected in the JHD mice, our results demonstrate that B cells play a critical role in CD4 T cell priming, possibly by delivering a critical co stimulatory activity for clonal expansion of CD4 T cells. PMID- 7495741 TI - Evidence for a calcium regulated, bidirectional intronic promoter in the murine TCR V alpha 1 gene. AB - Previous studies of the TCR alpha chain gene have located promoter elements 5' to the start of the various V alpha genes. The only fully characterized enhancer for the entire alpha chain gene (V, J and C genes) has been located approximately 3 kb from the 3' end of C alpha. We now report the existence of additional regulatory elements located in the introns of several murine V alpha genes (V alpha 1, V alpha 3 and V alpha B6.2.16). In the case of V alpha 1, this element appears to be a promoter with bidirectional activity that is not T cell specific. Interestingly, upstream of the promoter in the antisense strand, an open reading frame has been found that codes for a small molecular weight protein (approximately 60 amino acids) that contains a proline-rich region and a tyrosine isoleucine motif that has homology to Ig beta (the B29 gene product). A rabbit antiserum made against this sequence has confirmed its existence by Western blot and immunoprecipitation. Thus this V alpha 1 intronic promoter has the potential not only to induce the formation of a truncated V alpha 1 gene product, but also regulates the expression of a small molecular weight protein that may be involved in lymphocyte antigen receptor signaling. The activity of this promoter is regulated by changes in intracellular calcium. In the presence of ionomycin the promoter is down-regulated in the sense direction and its activity is enhanced in the antisense direction. This result suggests that this promoter can act differentially to produce two very different gene products. The bidirectional V alpha 1 promoter appears to be the first in the Ig superfamily to induce potentially functional proteins in both directions. PMID- 7495743 TI - Thymic selection events mediated by the pre-TCR do not depend upon a limiting ligand. AB - Thymocyte differentiation requires the production of a functional TCR, the culmination of a carefully orchestrated series of events in which TCR beta chain gene rearrangement precedes that of TCR alpha genes. The product of a successful rearrangement of the TCR beta locus associates with an invariant protein in immature thymocytes to form the 'pre-TCR' complex, which is required for allelic exclusion at the TCR beta locus, the expression of CD4 and CD8 co-receptors, and the clonal expansion of immature thymocytes. The pivotal role for the beta chain protein during early thymocyte development led us to investigate the relative differentiation efficiency within the same thymus of cells which do and cells which do not possess productive TCR gene rearrangements. Using mixed radiation bone marrow chimeras to establish an in vivo competition between TCR beta transgenic (Tg) and non-Tg bone marrow cells, we show that the prior productive rearrangement of a TCR beta chain gene only subtly enhances the efficiency of intrathymic differentiation. Further, we have compared the relative differentiation efficiency of TCR alpha beta and TCR beta Tg cells within the mixed chimera system by altering the proportion of TCR Tg bone marrow cells in the reconstituting inoculum. As expected, Tg cells carrying both alpha and beta chains of a selectable TCR are developmentally hindered compared with their non Tg counterparts by the lack of ample numbers of intrathymic positively selecting ligands or niches. In contrast, parallel experiments using TCR beta Tg bone marrow cells demonstrate that the early selection events mediated by the pre-TCR do not similarly depend upon a ligand present in limiting quantities. PMID- 7495744 TI - Functional activation of encephalitogenic T cells in the absence of antigen presenting cells. AB - Co-stimulatory signals provided by surface receptors of antigen-presenting cells (APC) are crucial for the activation of CD4+ T cells, classically measured by cell proliferation or IL-2 secretion. The contribution of APC co-stimulatory signals to the acquisition of various effector functions by activated T cells is not fully understood. We have now examined the importance of surface-mediated co stimulation by APC for activation of the effector potential of T cell clones mediating experimental allergic encephalomyelitis (EAE). We now report that T cell clones can be activated to produce EAE not only with APC but also by antibody-mediated TCR cross-linking in the presence of a mixture of T cell growth factors. Without activation, the T cell clones did not cause EAE. Therefore, at least some types of T cells can be activated to express their effector potential in the absence of any surface co-stimulatory signals requiring intact APC. PMID- 7495745 TI - Characterization of the non-functional Fas ligand of gld mice. AB - Mice homozygous for either the gld or lpr mutation develop autoimmune diseases and progressive lymphadenopathy. The lpr mutation is characterized by the absence of functional Fas, whereas gld mice exhibit an inactive FasL due to a point mutation proximal to the extracellular C-terminus. The structural repercussions of this amino acid substitution remain unknown. Here we report that FasL is expressed at similar levels on the surface of activated T lymphocytes from gld and wild-type mice. Using a polyclonal anti-FasL antibody, indistinguishable amounts of a 40 kDa protein are detected in both gld and wild-type splenocytes. The molecular model of FasL, based on the known structure of TNF-alpha, predicts that the Phe --> Leu gld mutation is located at the protomer interface which is close to the FasR interaction site. We conclude that the gld mutation allows normal FasL biosynthesis, surface expression and oligomerization, but induces structural alterations to the Fas binding region leading to the phenotypic changes observed. PMID- 7495746 TI - Cross-linking of the TCR-CD3 complex with CD4, CD8 or LFA-1 induces an anti apoptotic signal in thymocytes: the signal is canceled by FK506. AB - The immunosuppressant FK506 did not block differentiation of double-negative thymocytes into double-positive (DP) cells, but interfered with differentiation of DP cells into mature single-positive cells in a fetal thymus organ culture system, suggesting that FK506 inhibits positive selection. The drug also reduced the number of DP cells recovered after the culture. As positive selection depends on the inhibition of thymocyte apoptosis at its DP stage by signaling through the TCR-CD3 complex and some of the accessory molecules, including CD4, CD8 and LFA 1, we studied the possibility that FK506 enhanced apoptosis by itself or canceled the inhibition of apoptosis. The results indicated that FK506 was hardly toxic or hardly affected anti-CD3-induced DNA fragmentation in isolated thymocytes in vitro. On the other hand, upon cross-linking TCR-CD3 together with CD4, CD8 or LFA-1, FK506 markedly enhanced DNA fragmentation and cytolysis. The drug, however, hardly or only slightly enhanced these responses upon cross-linking TCR CD3 together with CD2, CD28, Thy-1 or H-2Kd. Cross-linking of TCR-CD3 together with CD4, CD8 or LFA-1 markedly inhibited glucocorticoid-induced death and the inhibition was canceled by FK506. Furthermore, cross-linking of TCR-CD3 together with LFA-1 potentially induces both an apoptosis-inducing signal and an FK506 sensitive anti-apoptotic signal, and that the latter signal may be related to an essential signal for positive selection. PMID- 7495747 TI - Kinetics of the reactions between the invariant chain (85-99) peptide and proteins of the murine class II MHC. AB - The region comprising residues 83-107 of the extracytoplasmic domain of the class II MHC-associated invariant chain protein is essential for its functional interaction with MHC proteins. A nested set of peptides that encompass this region, designated the class II invariant chain-derived peptides (CLIP), bind to many MHC proteins and inhibit the binding of antigenic peptides. The kinetics of the reactions between CLIP and five different murine class II MHC proteins have been determined. Specificity of CLIP binding was confirmed by competition with antigenic peptides. Large differences in the reaction rates were observed. For example, half-times of dissociation ranged from 4.4 min to 17.5 h, a > 200-fold difference. These results demonstrate that CLIP bind to MHC heterodimers at a site that involves the polymorphic residues. These data support the hypothesis that the CLIP binding site is within the peptide binding groove. It is further suggested that these differences in kinetic stabilities of CLIP-MHC protein complexes might affect the diversity of endogenous peptides bound to class II MHC proteins. PMID- 7495749 TI - Human naive T cells activated by cytokines differentiate into a split phenotype with functional features intermediate between naive and memory T cells. AB - We have recently shown that CD45RA+CD4+ naive T cells can be activated to proliferate by a combination of IL-2, TNF-alpha and IL-6, but, at variance with TCR-mediated activation, they do not acquire the CD45RO molecule. This prompted us to investigate the phenotype of these cells and the functional features they display upon TCR stimulation. Naive T cells expanded by cytokines, though remaining CD45RA+, express a variety of activation and adhesion molecules which are peculiar to effector or memory T cells. Naive cells primed by cytokines, when activated with anti-CD3 mAb, produce a broad spectrum of cytokines, express CD40 ligand, but are unable to help B cells for Ig synthesis. A subset of CD4+CD45RA+RO-T cells with a phenotype (HLA-DR-, VLA-2+ or IL-2R+) similar to that of cells activated by cytokines in vitro can be found in vivo. These results demonstrate that activation signals delivered by cytokines, in the absence of TCR stimulation, can activate naive T cells to proliferate and differentiate into a 'split phenotype' with elements common to both naive and memory T cells. This novel antigen-independent activation may help to maintain the naive T cell repertoire and facilitate the antigen-responsiveness of naive T cells. PMID- 7495748 TI - Differential regulation of IL-13 and IL-4 production by human CD8+ and CD4+ Th0, Th1 and Th2 T cell clones and EBV-transformed B cells. AB - In the present study, the requirements and characteristics for the production of IL-13 by human T cells, T cell clones and B cells were determined and compared with those of IL-4. IL-13 was produced by human CD4+ and CD8+ T lymphocyte subsets isolated from peripheral blood mononuclear cells and by CD4+ and CD8+ T cell clones. CD4+ T cell clones belonging to Th0, Th1-like and Th2-like subsets produced IL-13 following antigen-specific or polyclonal activation. In addition, EBV-transformed B cell lines expressed IL-13 mRNA and produced small amounts of IL-13 protein. Expression of IL-13 mRNA and production of IL-13 protein by peripheral blood T cells and T cell clones was induced rapidly and was relatively long lasting, whereas IL-4 production by these cells was transient. In addition, IL-13 mRNA expression was induced by modes of activation that failed to induce IL 4 mRNA expression. IL-13 shares many biological activities with IL-4 which is compatible with the notion that the IL-13 and IL-4 receptors share a common component required for signal transduction. However, IL-13 lacks the T cell activating properties of IL-4. Here we have shown that this is related to the fact that T cells fail to bind radiolabeled IL-13 and do not express the IL-13 specific receptor component. Taken together, these results indicate that the differences in expression and biological activities of IL-4 and IL-13 on T cells may have consequences for the relative roles of these cytokines in the immune response. PMID- 7495750 TI - A novel co-stimulatory T cell antigen co-expressed on renal cell carcinoma. AB - The A6H mAb raised primarily against human renal cell carcinoma (RCC) has previously been shown to bind strongly to RCC, to some degree to colon carcinoma but only marginally to a variety of normal tissues. Immunohistochemical analysis or RCC tissues containing tumor-infiltrating lymphocytes revealed that A6H stained both tumor cells and lymphocytes. FACS analysis of human peripheral blood cells demonstrated that A6H stained both tumor cells and lymphocytes. FACS analysis of human peripheral blood cells demonstrated that A6H mAb stained 85-90% of both CD4+ and CD8+ T cells, but not granulocytes, monocytes, NK cells or B cells. Furthermore, 85-90% of naive and memory T helper cells were stained with A6H suggesting that the A6H mAb defines unique subsets within these T cell populations. Dual staining showed that A6H mAb bind to an antigen that is clearly distinct from other cell surface molecules on T cells, including CD28, CD29, CD26, CD44 and ICAM-2. A6H mAb binding induced a second signal in anti-CD3 mAb activated T cells, resulting in cell proliferation, IL-2 receptor expression and vigorous production of IFN-gamma and TNF, and production of minor amounts of IL 2. Immunoprecipitation with A6H mAb indicated a molecular weight of 120-140 kDa on both T cells and RCC. We suggest that the A6H mAb defines a unique T cell surface antigen which is involved in signal transduction and is expressed on subsets of human T cells. The co-expression of A6H on T cells and tumor cells suggests a possible function related to common properties of these cells. PMID- 7495751 TI - Complement peptide C3a inhibits IgE-mediated triggering of rat mucosal mast cells. AB - The relationship between mast cells' secretory response to stimulation via their type 1 Fc epsilon receptors (Fc epsilon RI) and that provided by the C3a fragment of the complement system was investigated in the rat mucosal-type mast cell line RBL-2H3. These cells are known to be unresponsive to the so-called 'peptidergic' stimulus provided by cationic agents, such as anaphylatoxins, neuropeptides or polyamines. We now observed that C3a effectively inhibits the Fc epsilon RI clustering induced secretion of RBL-2H3 cells. This inhibition is dose-dependent and takes place at a C3a concentration range of 0.4-12.5 nM, i.e. at least three orders of magnitude lower than those where this anaphylatoxin exerts its secretory stimulus to 'serosal' mast cells. In order to identify where C3a interferes in the Fc epsilon RI coupling cascade, we have studied its effect on the cells' protein phosphorylation pattern, hydrolysis of phosphatidyl inositides, transient rise in free cytosolic Ca2+ ion concentration and Ca2+ uptake. All these processes were found to be inhibited by a similar C3a concentration range. PMID- 7495752 TI - Dysregulated expression of the IL-2 receptor beta-chain abrogates development of NK cells and Thy-1+ dendritic epidermal cells in transgenic mice. AB - The IL-2 receptor beta-chain (IL-2R beta), a specificity-determining subunit in the IL-2R complex with a restricted tissue distribution pattern, is essential for signal transduction. Our previous studies demonstrate that the continuous treatment of mice with anti-IL-2R beta resulted in the complete disappearance of NK cells and Thy-1+ dendritic epidermal cells (Thy-1+ dEC), suggesting that signals through IL-2R beta are critically involved in development of these lymphocyte subsets. However, these lymphocyte subsets are reported to be apparently unaffected in the IL-2-deficient mice. To further examine the biological roles of the IL-2R beta, transgenic mice carrying the IL-2R beta transgene were generated. In these mice, high levels of the cell surface expression of the IL-2R beta were observed in essentially all hematopoietic lineage cells, and CD4+ T cells as well as CD8+ T cells showed vigorous cell proliferation upon IL-2 stimulation. Surprisingly, NK cells marked with a high expression of NK1.1 in the spleen and Thy-1+ dEC in the skin were completely absent in transgenic mice. However, the development of other lymphocyte subsets including conventional alpha beta TCR+ cells, gamma delta TCR+ cells and B cells remained apparently intact. From these observations together with previous data on IL-2-deficient mice, we speculate that factors, other than IL-2 that utilizes the IL-2R beta as its functional receptor subunit, may have a vital role in the development of NK cells and Thy-1+ dEC. Implications for possible in vivo functions of over-expressed IL-2R beta are discussed. PMID- 7495753 TI - Fas (CD95) participates in peripheral T cell deletion and associated apoptosis in vivo. AB - Following exposure to some types of antigen (superantigens), responsive T cells expand and then decline in numbers, a phenomenon that has been called 'peripheral deletion'. This process may play a role in limiting autoimmune reactions and in the maintenance of immune homeostasis. Here we describe experiments on peripheral deletion in mice carrying the lpr/lpr defect, which has been shown to be due to defective production of the CD95/Fas molecule. Young lpr/lpr mice with no apparent immunologic abnormalities display a defect in bacterial superantigen induced peripheral deletion. Apoptotic death of the expanded T cell population associated with such peripheral deletion. Apoptotic death of the expanded T cell population associated with such peripheral deletion in normal animals is dramatically reduced in the mutant mice. Further, the levels of Fas on responding cells in normal mice increases and decreases together with increases and decreases in cell numbers, suggesting that cells with the highest levels of Fas are preferentially deleted. These observations are consistent with the known ability of CD95 to transduce a signal leading to apoptosis, and they implicate this signal transduction pathway in peripheral deletion. In contrast, bacterial superantigen-induced deletion of thymocytes appears to be fully functional in these mice, and thus Fas/APO-1 does not appear to be required for this process. Further, antibody ligation of the TCR on activated T cells from normal or young lpr/lpr mice can induce apoptosis and therefore under some circumstances this phenomenon is not dependent upon CD95/Fas. Thus, to avoid autoreactivity and ensure immune homeostasis, several different apoptotic mechanisms exist in peripheral T lymphocytes, only some of which involve Fas.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7495754 TI - Inactivation of E alpha and E beta expression in inbred and wild mice by multiple distinct mutations, some of which predate speciation within Mus species. AB - The H-2 MHC of mice encodes two functional class II heterodimeric proteins: A alpha A beta (A) and E alpha E beta (E). While failure to express the A protein has not been reported, a significant proportion of of H-2 haplotypes in both inbred and wild mice do not express E proteins. We and others have previously characterized the molecular basis for defective E expression in haplotypes from Mus domesticus (b, f, q, s, from inbred strains) and M. castaneus (w17, wild derived) species, identifying six distinct defects in the genes for E alpha or E beta. In this report we have extended these studies to other E- haplotypes, including several from t-haplotype-bearing M. domesticus mice (w29, w57, w302) and one derived from the Asian species M. bactrianus (w301). Analyses at the protein, RNA and DNA levels were employed to identify the defects in the genes for Ea and Eb. At least one new defect was identified that prevents E beta expression in a t-associated H-2 haplotypes (w57), bringing the number of distinct mutations causing the E- phenotype to seven. Another t-associated haplotype, w302, was found to share the same E beta defect with mice of the inbred q haplotype and of the w17 haplotype from M. castaneus, while its Ea gene contains the deletion carried also by the inbred b and s haplotypes and by a number of wild haplotypes. The mutations in the Ea and Eb genes of the w301 haplotype from M. bactrianus were found to be identical to those of the inbred f haplotype. This indicates that the origin of the mutations in the Eb genes of the q, w17 and w302 haplotypes and in the Ea and Eb genes of the f and w301 haplotypes, predated speciation within Mus, thought to have occurred approximately 0.35-1 million years ago. Their maintenance in mouse populations suggests that in certain conditions the failure to express E alpha E beta proteins may be advantageous and selected for. PMID- 7495755 TI - Intra-epithelial lymphocytes. Evidence for regional specialization and extrathymic T cell maturation in the human gut epithelium. AB - The human gut epithelium is a unique immunological compartment, containing substantial amounts of intra-epithelial lymphocytes (IEL) with unknown functions. In this study we show that distinct and unusual subpopulations of IEL are present at different levels of human intestine. IEL phenotypes in normal jejunum, ileum and colon were compared using immunoflow cytometry and immunohistochemistry. The expression of mRNA for recombination-activating gene-1 (RAG-1) in IEL from all three levels was compared using reverse-transcription polymerase chain reaction, and the morphology of IEL in situ was determined using immunoelectron microscopy. Surface marker profiles of isolated intestinal epithelial cells at all three levels were also investigated. On average the proportion of TCR gamma delta IEL was comparable in jejunum than ileum and colon and varied in phenotype with gut level. CD4-CD8-TCR alpha beta IEL dominated in colon but were absent in jejunum. CD8+ TCR alpha beta IEL were present at all levels but only in jejunum did they constitute the majority of all IEL. CD4+ TCR alpha beta IEL were present in similar frequencies at all levels of the gut. In general, the majority of IEL had an activated phenotype (CD45RO+, alpha E beta 7+). Furthermore, IEL exhibited phenotypes which are rare in peripheral blood. The thymocyte markers CD1a and CD1c as well as the NK cell marker CD56 were expressed on a fraction of TCR alpha beta and TCR gamma delta IEL. A small population of 'null' cells (CD45+ TCR/CD# CD20-CD14-CD15- cells) was also present at equal proportions along the gut. Jejunal but not colonic IEL expressed RAG-1 mRNA suggesting that extrathymic T cell maturation occurs in the epithelium of small intestine. RAG-1 was expressed in CD2+TCR/CD3- and CD3+/TCR-IEL. Ultrastructurally, IEL often formed small clusters and intimate contacts with epithelial cells, suggesting cell cooperation within the epithelium. Some IEL had pseudopodium-like extensions penetrating the epithelial basement membrane suggesting transmigration. Epithelial cells in small intestine but not colon expressed heat shock protein 60 and HLA-DR. CD1a, CD1b and CD1c were not expressed on intestinal epithelial cells at any level. The distinct surface marker profiles of IEL and epithelial cells along small and large intestine suggest functional regional specialization and are compatible with the hypothesis that TCR alpha beta IEL participate in immune reactions to lumenal antigens while TCR gamma delta IEL perform surveillance of the epithelium. PMID- 7495756 TI - Activation of a myelin basic protein-specific human T cell clone by antigen presenting cells from rhesus monkeys. AB - This study addresses the capacity of peripheral blood mononuclear cells (PBMC) from rhesus monkeys (Macaca mulatta) to present myelin basic protein (MBP), a candidate auto-antigen for multiple sclerosis, to MBP-specific human CD4+ T cell clones. MHC-restriction of the human T cell clones was determined with HLA-DR transfected L cells, and epitope specificity was established with a panel of overlapping 20-mer peptides. The MHC-DR region of the rhesus monkeys (Mamu) was characterized serologically and by sequence analysis. We identified one CD4+ HLA DRB1*0301-restricted Th1-like human T cell clone (ES-BP8) that was activated to proliferation with human or rhesus monkey MBP, or peptide MBP 29-48 presented by PBMC from six different rhesus monkeys expressing the Mamu-DRB1*0305 or DRB1*0306 alleles. After transformation to continuous growth with Herpesvirus saimiri, the T cell clone could still be stimulated by antigen (Ag) and Ag presenting cells (APC) from monkeys. Two other T cell clones with the same HLA restriction and the same peptide-specificity did not respond to MBP presented by these rhesus monkeys. The exon 2 sequences HLA-DRB1*0301, Mamu-DRB1*0305 and DRB1*0306 differ at positions 32, 47, 67, 73 and 86. These amino acid differences are not critical for the binding of MBP 29-48 and do not abrogate recognition by the clone ES-BP8, but interfere with the recognition of the two other HLA DRB1*0301-restricted T cell clones. In conclusion, studying Ag-presentation from rhesus monkey may provide further insight into the interaction of antigenic peptide, TCR and MHC.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7495757 TI - Positive and negative regulation of Thy-1 expression on B lymphocytes by IL-4. AB - Expression of the Thy-1 membrane antigen is generally confined to thymocytes and T lymphocytes, but its expression on B lymphocytes can be induced by culture with the lymphokine IL-4. IL-4 was first reported as a soluble factor capable of participating in the activation of B cells. However, it has been shown that the proliferative response of B cells to IL-4 is dependent upon both their stage of differentiation and their prior exposure to other activating signals--under some conditions, IL-4 can inhibit B cell functions. The present study was designed to determine whether IL-4 signaling induces Thy-1 expression on all B lymphocytes, or whether this induction is dependent upon IL-4-mediated activation. We examined the role of IL-4 in regulating both mRNA and protein levels of Thy-1 in three mouse B cell lines with distinct growth responses to IL-4. IL-4 was required for Thy-1 expression in cells which were dependent upon IL-4 for continuous growth in culture but markedly decreased Thy-1 expression in cells which are growth inhibited by IL-4. In a mutant subclone of the latter cells in which IL-4 signaling does not cause growth inhibition, IL-4 did not affect Thy-1 expression. The regulation of Thy-1 expression by IL-4 is manifest at the level of Thy-1 mRNA. Thus, IL-4 can both positively and negatively influence B cell expression of Thy-1, depending on the growth response of the cells to IL-4. PMID- 7495758 TI - The primary alloresponse of human CD4+ T cells is dependent on B7 (CD80), augmented by CD58, but relatively uninfluenced by CD54 expression. AB - Conflicting data have been reported regarding the relative abilities of B7, ICAM 1 and LFA-3 to provide co-stimulation for the induction of a primary T cell alloproliferative response. A series of naturally HLA-DR-expressing cell lines and panels of human and murine transfectants expressing DR alloantigens in conjunction with combinations of mouse or human B7.1, human LFA-3 and human ICAM 1 were used to analyse the contributions of these molecules to primary alloproliferative responses by adult and cord blood CD4+ T cells. The results demonstrated that B7 expression is required, and may be sufficient for the induction of a primary alloresponse. The allostimulation observed in response to DR-expressing murine DAP.3 cells, that constitutively express B7.1, was inhibited by the presence of the murine cytolytic T lymphocyte-associated antigen 4-human Fc gamma 11 fusion protein, suggesting that mouse B7.1 provides sufficient costimulation for a primary human alloproliferative response. Expression of supranormal levels of human B7.1 on the allostimulator cells led to a reduction in the proliferative response, suggesting that an optimal level of B7 exists which, if exceeded, leads to inhibition. Co-expression of LFA-3 with B7.1 by the allostimulator cells caused a marked increase in the proliferative response. Expression of ICAM-1a had relatively little effect. No differences were seen in the co-stimulatory requirements of naive cord blood versus CD45RO adult T cells. There results highlight the key molecular interactions that govern immunogenicity with relevance to inhibiting unwanted immune response to transplanted tissues and provoking anti-tumour immunity. PMID- 7495759 TI - CIITA activates the expression of MHC class II genes in mouse T cells. AB - It has long been a puzzle that MHC class II molecules are expressed in human T cells after activation but not in mouse T cells; this expression is believed to play a role in the cell mediated immune response. Recently the MHC class II transactivator (CIITA) has been reported to be a major regulatory factor for both the constitutive and IFN inducible expression of MHC class II genes. Here we show that human T cells expressing MHC class II have CIITA transcripts while MHC class II-negative human T cells and mouse T cells do not. The expression of MHC class II genes in mouse T cells can be reconstituted upon transfection with the human CIITA cDNA. These data indicate that the expression of CIITA explains the expression or lack of expression of MHC class II in human and mouse T cells respectively. PMID- 7495760 TI - Interleukin 12 alters the isotype-restricted antibody response of mice to hen eggwhite lysozyme. AB - Protein antigens elicit humoral responses in mice that consist predominantly of IgG1 antibodies. We have now investigated the ability of IL-12, a cytokine reported to augment IgG2a anti-hapten responses through activation of Th1 cells, to alter antibody responses to hen eggwhite lysozyme (HEL). The normal response of BALB/c mice to HEL is highly restricted to IgG1 expression and therefore provides an excellent system for determining effects of cytokines on expression of other isotypes. Seven days after immunization, IL-12 treated mice demonstrated greatly elevated HEL-specific IgG2a antibody levels and suppressed IgG1 production, while PBS-treated control mice showed a typical IgG1-restricted response. On day 28, IL-12-treated mice showed heightened serum antibody levels of both isotypes. Delaying cytokine treatment until after the typical IgG1 anti HEL response had already been established also led to significant elevation of serum IgG2a antibody levels. These effects correlated with increased IFN-gamma production; however, administration of IL-12 plus anti-IFN-gamma had little influence on IgG2a enhancement, although it did relieve the early IgG1 suppression. Furthermore, the differential effects of Il-12 on isotype expression did not correlate with time; in fact, IgG2a enhancement correlated with loss of IgG1 suppression. Our findings indicate that (i) IL-12 reproducibly induces large amounts of IgG2a HEL-specific antibodies in vivo; (ii) it can alter isotype profiles of both primary and secondary responses; and (iii) its effects on humoral immunity are not completely explained by induction of Th1 cell derived IFN-gamma. PMID- 7495761 TI - Evidence for a CD40 response element, distinct from the IL-4 response element, in the germline epsilon promoter. AB - Engagement of CD40 by its ligand induces transcription of unrearranged Ig heavy chain genes, an initial step in switch recombination. The following studies were undertaken to understand the molecular basis of this response. Co-culture of S19 cells expressing membrane-bound CD40 ligand (CD40L) encoded by recombinant baculovirus with EBV-transformed B cell lines induced germline transcription of the epsilon gene in the absence of cytokines. To identify a putative CD40 response element, a reporter construct consisting of the 777 bp of the 5' flank of the human l epsilon region linked to the chloramphenicol acetyl transferase (CAT) gene was stably transfected into B cell lines. Stimulation with either CD40L-expressing Sf9 cells or IL-4 induced CAT activity. Deletional analysis of this promoter region confirmed that an IL-4 response element was identified within a 63 bp segment 3' to the IL-4-responsive element that was responsive to CD40 ligation. These results indicate that the germline epsilon promoter contains a CD40 response element that is distinct from that accounting fro IL-4 responsiveness. Activity of this response element may explain the capacity of ligation of CD40 to induce germline epsilon transcripts in the absence of cytokines. PMID- 7495762 TI - New hydrophilic polyesters and related polymers as bioerodible polymeric matrices. AB - A survey is reported on our activity performed in the last few years on the preparation of new synthetic and semisynthetic polymeric materials endowed with bioerodible-biodegradable characteristics and designed for applications in the practice of controlled release of active principles of pharmaceutical and agrochemical significance. The presentation of the results will be arranged into the following sections: (1) hydroxyl containing polyesters, that comprise polymerization products based on racemic and optically active glyceric acid, or attained by polyaddition reactions among cyclic anhydrides, including also carbon dioxide, with monoglycidyl ethers of reversibly protected polyols. In this class are also presented the related polyhydroxylated systems obtained by selective grafting functional epoxides on cyclodextrins. (2) Bioerodible carboxyl containing polymeric systems as derived from the alternating copolymerization of maleic anhydride with alkyl vinyl ethers followed by partial esterification of maleic anhydride groups. (3) Linear and cross-linked functional polymers of synthetic and semisynthetic origin with hydrogel forming capability. Typical examples of their applications in the release of drugs and phytodrugs are also presented. PMID- 7495763 TI - Phagocytosis of monosize polystyrene-based microspheres having different size and surface properties. AB - In this study, nondegradable monosize polystyrene (PS) based polymeric microspheres with different size and surface chemistries were prepared by different polymerization techniques. Surfaces of the plain microspheres were further modified biologically by albumin (BSA) or fibronectin (Fn) preadsorption. Phagocytosis of these polymeric microspheres by leukocytes and macrophages were investigated. The phagocytic response of both leukocytes and macrophages decreased by increasing size of the particles. More hydrophilic particles phagocytosed less. Positive charges increased the uptake while negative charges oppositely reduced the uptake. BSA on the surface almost prevented the uptake, while Fn caused opsonization. PMID- 7495764 TI - Transplantation of microencapsulated hepatocytes for liver function replacement. AB - Recent advances in cell biology and biotechnology have lead the way for a greater understanding of cell function and the potential therapeutic use of transplanted cells for treating a wide array of illnesses. Treatment of disease by transplantation of normal healthy cells, for the replacement of specific biological deficiencies or as a form of auxiliary support for a failing organ, offers important therapeutic applications and also serves as a model for assessing cellular physiology. In the long-term, cell transplantation may also have potential in the development of artificial organ support systems for sustaining patients with severe and chronic diseases such as diabetes, liver failure, endocrine and exocrine disorders, neurological abnormalities, and congenital metabolic defects. Several groups have demonstrated the feasibility and efficacy of cell transplantation in providing specific function in various experimental animal models of human disease. However, without adequate immunosuppression, complications due to tissue rejection remain a significant problem. Microencapsulation of cells within a synthetic semipermeable membrane, prior to transplantation, has been proposed for circumventing immunological complications following transplantation. The microcapsule's semipermeable membrane allows permeant molecules to freely diffuse across while preventing the microencapsulated cells from escaping. This membrane also keeps unwanted substances, such as cells and antibodies, from entering the microcapsule. Thus, microencapsulation provides an innovative and unique technique for the transplantation of foreign tissue and cells without the need for immunosuppression. PMID- 7495765 TI - Novel PDLLA/PEG copolymer micelles as drug carriers. AB - In this study we attempted to develop a novel drug delivery system in the form of polymeric micelles. In order to obtain polymer chains with micelle forming abilities, we have proposed to produce copolymers by transesterification of poly(DL-lactic acid)(PDLLA)(as the hydrophobic segment) and polyethylene glycol (PEG) (as the hydrophilic segment). We first produced homopolymers of PDLLA with low molecular weights by condensation polymerization of DL-lactic acid. The PDLLA homopolymer with an average molecular weight of 1866 +/- 100 was then transesterified with PEG with a molecular weight range of 3300-4000. By changing the ratio of PEG to PDLLA, we were able to produce copolymers with different chain structures, and therefore, with different micelle forming abilities. FTIR, 1H-NMR, DSC, and GPC studies were performed to describe the structures of these PDLLA/PEG copolymers. Micelles of these copolymers were characterized by light scattering. We selected a model drug, i.e. adriamycin, and obtained the drug loadings to the PDLLA/PEG copolymer micelles. The maximum drug loading was about 12 mg g-1. We found that these micelles were degraded in phosphate buffer (pH 7.4 and temperature 37 degrees C) in about 5-6 weeks. We also investigated the release of adriamycin from these PDLLA/PEG micelles, and concluded that the drug release from these micelles was mainly 'degradation controlled'. PMID- 7495766 TI - Orientruded polylactide based body-absorbable osteosynthesis devices: a short review. AB - A new process 'orietrusion' has been developed to impart a high degree of molecular orientation into polylactide. Mechanical tests showed that this process increased the initial strength of the high molecular weight polylactide by 690%. Orthopaedic osteosynthesis devices that were made from this oriented polyactide were shown to be biocompatible and body-adsorbable. Screws that were made from this material were shown to be efficacious in canine calcaneous and femoral osteotomy models. The orientruded polylactide screws were shown to be efficacious in the fixation of the human malleolar fractures. PMID- 7495767 TI - Frequent reactivations of anti-HBe-positive chronic hepatitis B in patients with no demonstrable HBV DNA in serum by polymerase chain reaction. AB - OBJECTIVES: The present study was conducted to evaluate the prognostic significance of the absence of serum HBV DNA by polymerase chain reaction (PCR) after spontaneous HBeAg/anti-HBe seroconversion and concurrent or subsequent biochemical remission. METHODS: We prospectively investigated the reactivation rates in 28 chronic hepatitis B patients according to the positive or negative serum HBV DNA test by PCR. The sera drawn at a mean period of 4.4 months after normalization of alanine aminotransferase (ALT) were analyzed by PCR-Southern blot hybridization to detect HBV DNA, and then the patients were divided into two groups according to the presence (n = 14) or absence (n = 14) of HBV DNA in the sera. RESULTS: The cumulative reactivation rates in patients with HBV DNA in sera were 43%, 57%, 57%, 57% and 57% at the end of 1st, 2nd, 3rd, 4th and 5th year after normalization of ALT, respectively, and those in patients without demonstrable HBV DNA were 50%, 66%, 74%, 74% and 83%, respectively; thus, the difference in the cumulative reactivation rates between patients with and without serum HBV DNA was not statistically significant (p = 0.79), and irrespective of the status of HBV DNA in sera by PCR, reactivations occurred very rarely after 2 years of a sustained remission. CONCLUSIONS: We conclude that the seroconversion to anti-HBe accompanied by disappearance of serum HBV DNA even by PCR does not necessarily suggest a sustained remission of chronic hepatitis B. PMID- 7495768 TI - A new method of endoscopic variceal ligation-injection sclerotherapy (EVLIS) for gastric varices. AB - OBJECTIVES: For esophageal varices, endoscopic variceal sclerotherapy and band ligation have been effectively and frequently used, but these methods were not effective for gastric varices. In gastric varices, because of rich networks of feeding vessels and shunts that may exist in many cases, an effective therapeutic level of sclerosant is difficult to be maintained. Accordingly, we propose a new method called "Endoscopic Variceal Ligation-Injection Sclerotherapy (abbreviated EVLIS)" for gastiric varices, with focus on maintenance of the effective therapeutic level of a sclerosant by partially blocking the blood flow of gastric varices using the band ligation method. The study was undertaken prospectively to evaluate the efficacy and safety of EVLIS for the treatment of gastric varices. METHODS: EVLIS was performed in a group of 32 patients with gastric varices. Active bleeding varices were 11 and non-bleeding 21. Five cases were grade A, 12 grade B, and 14 grade C of Child's classification. Nine cases were type 1, 22 type 2, and 1 was type 3 of Hosking-Johnson's classification. RESULTS: The results of EVLIS were excellent, active bleeding of gastric varices in the 11 cases were successfully controlled and all the gastric varices of 32 cases including those bleeding varices were completely eradicated. The bleeding status, the Child's grade and the Hosking-Johnson's class do not appear to correlate in any way with the efficacy of this technique. No perforation or re-bleeding was observed in any of the patients until the mean 10.6 months of the follow-up period. CONCLUSIONS: EVLIS should be considered as an effective and safe treatment for gastric varices. PMID- 7495769 TI - Effects of endoscopic variceal ligation in lower esophageal motor function: a prospective study. AB - OBJECTIVES: Endoscopic variceal ligation (EVL), a recently developed method for controlling active variceal bleeding and eradicating esophageal varices, has similar efficacy to endoscopic injection sclerotherapy (EIS) and is known to have a minimal risk of complications and fewer complications in the lower esophagus. However, since the site of EVL is chiefly done in the lower esophagus, we prospectively evaluated to investigate the effect of EVL on the lower esophageal motor function. METHODS: We evaluated the severity of esophageal varix with the endoscopy and the lower esophageal manometry in 27 patients who had no history of interventional therapy, for varices before EVL, 3 weeks and 6 months after the last EVL session. RESULTS: The EVL caused considerable diminution in the size of esophageal varix by a mean 8.2 (range 3-21) ligations in mean 1.7 (range 1-3) sessions. In most of the cases, the varices reappeared and enlarged when the procedure of EVL was stopped. There were two different types of changes (intermediate and late) in the lower esophageal motility. The intermediate post EVL effects were the increase of peristaltic contraction amplitude and duration in the lower esophageal body after EVL. The late post-EVL effects were the prolongation of lower esophageal sphinctor (LES) relaxation duration and speedier peristaltic velocity in the lower esophageal body. CONCLUSIONS: We conclude from these findings that the intermediate post-EVL effect may be transient and the increase of peristaltic wave was due to diminution of esophageal varix. PMID- 7495770 TI - Influence of Helicobacter pylori colonization on histological grading of chronic gastritis in Korean patients with peptic ulcer. AB - OBJECTIVES: We conducted an analysis of correlation between histological grading of chronic gastritis and the presence of H. pylori infection to investigate if H. pylori influences histological severity of chronic gastritis in Korean patients with peptic ulcers. METHODS: Gastroscopic antral biopsy specimens and peripheral venous blood were taken from 80 patients with gastric or duodenal ulcers. H. pylori was identified microscopically in sections with Giemsa staining and quantitative grading of cultured H. pylori was reported on a scale 0 to 3. The histopathological features of biopsy specimens were reported according to the Sydney classification of chronic gastritis. Serum gastrin and pepsinogen concentrations were measured by radioimmunoassay. RESULTS: H. pylori was identified in 62.5%(20 of 32 GU, 30 of 48 DU) of the study group. Gastric clonization rate of H. pylori did not increased with age. Forty of 50 biopsy specimens with H. pylori and also 23 of 30 biopsy specimens without H. pylori showed active chronic gastritis. There was no significant correlation overall between the presence of H. pylori and histological grading of chronic gastritis, including activity, and also no association was found between the quantitative grading of H. pylori and the histological grading of chronic gastritis. With and without H. pylori, a mean of serum gastrin concentration(79.4 +/- 4.30 pg/ml and 80.2 +/- 31.9 pg/ml)showed no significant difference, but a mean of serum pepsinogen concentration(87.7 +/- 41.6 ng/ml and 119 +/- 34.4 ng/ml) showed significant difference between the populations with and without H. pylori(p = 0.001). CONCLUSIONS: The influence of H. pylori on histological grading of chronic gastritis in Korean is less than that in prior studies of Western countries, and further investigation of pathogenesis of H. pylori in chronic gastritis and peptic ulceration is necessary. PMID- 7495773 TI - Evolution of left atrial thrombus with anticoagulant therapy-follow-up by transesophageal echocardiography. AB - OBJECTIVES: Atrial fibrillation is an important risk factor for systemic embolism. A number of clinical studies demonstrated the beneficial effect of anticoagulant therapy for the prevention of embolism. But there has been no study on the fate of left atrial thrombus demonstrated by transesophageal echocardiography in the course of anticoagulation therapy. METHODS: Thirteen patients, demonstrated to have left atrial thrombus by transesophageal echocardiography were followed with anticoagulation therapy. Repeated transesophageal echocardiography was done 15 months after 1st study. RESULTS: Among 9 patients with adequate anticoagulation effect (INR > 2.0), left atrial thrombus disappeared in 3 patients. The size of thrombus decreased from 2.2 +/- 0.8cm to 0.9 +/- 1.0cm (p < 0.05 by paired Student's t-test). CONCLUSIONS: Left atrial thrombus could dissolve or decrease in size with adequate anticoagulation. PMID- 7495772 TI - PR interval behavior during exercise stress test. AB - OBJECTIVES: The PR interval on the electrocardiogram represents conduction time from the onset of atrial activation through His-Purkinje conduction system during a normal cardiac cycle. While its behavior at rest and during exposure to various cardioactive drugs is well documented, there exist few reports which describe PR interval variations during exercise in normal control and patient groups. In the present study, we examined the behavior of the PR intervals during various stages of exercise, and at the same time we observed whether the changes of PR interval during exercise could suggest that implanted cardiac pacemaker algorithms may be constructed to maximize hemodynamic benefits in patients requiring physiological cardiac pacemaker. METHODS: A retrospective analysis of the exercise treadmill test was performed on 148 healthy control group (148 males, mean age of 42.7 +/- 11.7) and 134 patient group (95 males & 39 females, mean age of 47.1 +/- 11.7) which had complained of non-specific chest symptoms but were identified as normal in the exercise treadmill test. During the test, we used the standard Bruce protocol. The results were expressed as mean +/- standard deviation, and differences in the mean value of each standard deviation, and differences in the mean value of each group were evaluated by the student's t-test. A P value of less than 0.05 was regarded as significant. RESULTS: 1) The control group showed 6.9msec reduction rate of the PR interval whenever their heart rate increased by 10 beats per minute. 2) The entire patients group showed 5msec reduction rate of the PR interval whenever their heart rate increased by 10 beats per minute. 3) The male patients group showed 5.2msec reduction rate of the PR interval whenever their heart rate increased by 10 beats per minute. 4) The female patient group showed 4.3msec reduction rate of the PR interval whenever their heart rate increased by 10 beats per minute. 5) There were significant differences of the PR interval changes between the entire or male patient group and the control group within the same range of heart rates. CONCLUSIONS: This study shows that PR interval changes corresponding to heart rate increments were linearly decreased. These changes of PR interval during exercise suggest that implanted cardiac pacemaker algorithms may be constructed to maximize hemodynamic benefits in patients requiring physiological cardiac pacemakers. PMID- 7495771 TI - TNFB gene polymorphism in patients with systemic lupus erythematosus in Korean. AB - OBJECTIVES: To elucidate the gene frequency of TNFB Ncol polymorphism and its association with HLA class II antigen in patients with systemic lupus erythematosus(SLE) in Korea. METHODS: We investigated the gene frequency of the TNFB alleles using DNA obtained from peripheral mononuclar cells in 141 healthy controls and in 58 patients with SLE. The polymorphisms of TNFB gene (735 bp) were studied by Ncol PCR-RELP. A portion of TNFB gene(735 bp) was amplified by PCR and its products were digested with Ncol restriction enzyme. The digested samples of amplified DNA were analyzed by agarose gel electrophoresis. TNFB*1 and TNFB*2 alleles were identified according to polymorphic fragments on Ncol restriction site in the first intron of the TNFB gene. The generic types of HLA DRBI were also determined by PCR with sequence specific primers(SSP) using genomic DNA from the same subjects. RESULTS: The genotypic frequency of TNFB*2 homozygote was significantly increased in patients with SLE compared with controls(RR = 2.36, P = 0.011). The frequency of HLA-DRBI*15 was also significantly increased in patients (RR = 2.27, P = 0.029). However, the increased frequency of TNFB*2 homozygote was apparently increased in nephritis group (RR = 2.79, P = 0.035), whereas the significance of TNFB*2 homozygote was weakend in non-nephritis group. CONCLUSIONS: Our results suggest that genetic predisposition of TNFB*2 homozygote is another risk factor in Korean SLE, especially in DR2 negative patients. In addition, TNFB*2 homozygote could have a tendency for the development of nephritis in patients with SLE. PMID- 7495774 TI - Bacteremia with nontyphi Salmonella and therapeutic implication. AB - OBJECTIVES: The recent decades witnessed the increased incidence of bacteremia with nontyphi Salmonella, especially in patients with underlying diseases. To evaluate clinically the patients with bacteremia by nontyphi Salmonella, we investigated the clinical records of the patients. METHODS: Retrospective study was performed on 30 cases of bacteremia with nontyphi Salmonella admitted to Soon Chun Hyang University Hospital from 1987 to 1993, and we analyzed with regard to age distribution, underlying diseases, clinical presentation and fatality. RESULTS: Children below 2 years of age were seven patients and patients over 55 years of age were 8 patients. The male to female ratio was 3:2. Only five cases were nosocomially acquired. More than two-thirds (21 patients) had no discernible other illness at the time of their bacteremia. A quarter of the patients presented without gastrointestinal manifestation. There were four deaths in the 30 patients with nontyphi Salmonella septicemia (13.3%). CONCLUSIONS: We investigated the clinical characteristics of 30 cases of non-typhi Salmonella bacteremia, which has a relatively high incidence nowadays while, to the contrary, Salmonella typhi trends toward being on the decrease in Korea. PMID- 7495775 TI - A case of massive cisplatin overdose managed by plasmapheresis. AB - OBJECTIVES: Accidental cisplatin overdose occurs with increasing frequency despite the safeguards taken in prescription and administration, since cisplatin has been used increasingly for the treatment of numerous malignancies. Accidentally, a 59-year-old male received massive cisplatin overdose of 300mg/m2. METHODS: Laboratory documentation included measurement of cisplatin concentrations by flameless atomic absorption spectroscopy (Varian, Spectra AA 300). RESULTS: Toxicities included severe emesis, myelosuppression, renal failure, mental deterioration with hallucination, dim vision and hepatic toxicity. Plasmapheresis was effective in lowering the platinum concentration from greatest 2979 ng/ml to 185 ng/ml and appeared to be of clinical benefit. Granulocyte-macrophage colony stimulating factor (GM-CSF) was used to ameliorate myelosuppression. The patient's renal function was restored 3 months later and partial response of esophageal cancer was obtained. CONCLUSIONS: Plasmapheresis was effective in lowering the platinum concentration in massive cisplatin overdose. This case heightens awareness to the possibility of accidental cisplatin overdose and the benefits of prompt management. PMID- 7495777 TI - A case of granulocytic sarcoma of the brain in a patient with myelodysplastic syndrome. AB - Granulocytic sarcoma is rare extramedullary tumor composed of myeloblasts and other granulocytic precursors. The majority of cases have been reported in association with acute myeloid leukemia (AML) or myeloproliferative disorders. Granulocytic sarcoma may occur in patients with myelodysplastic syndromes. Reports are very rare, especially in the brain. We report an unusual case of granulocytic sarcoma of the parenchyma of the brain, occurring in a patient with myelodysplastic syndrome, diagnosed by cerebro-spinal fluid cytology and magnetic resonance imaging brain scan. PMID- 7495778 TI - Long-term therapy of hypertension. PMID- 7495779 TI - Association of the core clustering mutations (codon 21-34) and the severity of chronic hepatitis B in Korean patients. AB - OBJECTIVES: There are regions in the core gene of hepatitis B virus (HBV) where missense mutations are clustered, and mutations in that region are related to severe liver disease. However, there were some differences of the major regions for mutation clustering among ethnic groups. To explore the phenomenon of clustering mutations in Korean patients with chronic HBV infection and to elucidate the correlation between clustering mutation region of the core gene and the severity of liver damage, we analyzed the precore/core gene sequence of HBV in the sera from fifteen chronic hepatitis B (CH-B) patients. METHODS: We analysed the HBV precore and core sequences in the sera obtained from fifteen patients (14 males and 1 female, mean age 30.0 years) with biopsy-proven CH-B. The patients were divided into two groups according to the pathological severity of CH-B; namely, group I consisted of 8 patients with chronic persistent hepatitis (CPH), and group II included 7 patients with chronic active hepatitis (CAH). After extraction of HBV DNA from each serum by proteinase K and phenol chloroform solution, the entire precore and core region of HBV was amplified by PCR, and then the PCR products were subjected to direct sequencing using thermostable DNA polymerase. Fisher's exact test and Mann-Whitney U test were used for statistical analysis. RESULTS: A total of 181 nucleotide substitutions were found in the HBV core gene from the 15 CH-B patients, of which 23 were missense and 158 were silent. The nucleotide and amino acid substitution rates were not significantly different between the two groups (p > 0.05). Two mutational hot spots (MHS), codons 21-34 (MHS1) and codons 85-100 (MHS2), were found in the deduced amino acid alignment of the core gene. The alteration rate of amino acid residue in these regions were 2.857 x 10(-2) and 5.000 x 10(-2), respectively. Of 8 CPH patients, 5 showed missense mutations only in MHS2. In comparison, of 7 CAH patients, 3 showed them both in MHS1 and MHS2, 1 only in MHS1, and 1 only in MHS2; thus, missense mutation in MHS1 was exclusively found in patient with CAH. CONCLUSIONS: There were two mutation clusterings in the core region of adr subtype of HBV from Korean CH-B patients. Mutations in MHS1 (codon 21-34), but not in MHS2 (codon 85-100), are more likely to be related to the severity of CH-B. A longitudinal study using sequential samples is warranted to further clarify the role of MHS1 in the pathogenesis of more severe CH-B. PMID- 7495776 TI - A case of amiodarone-associated pulmonary toxicity. AB - Amiodarone is an iodinated benzofuran derivative that represents a new and extremely effective therapy for certain life-threatening refractory cardiac arrgythmia. There are numerous side effects associated with amiodarone therapy, including corneal microdeposits. Skin reactions and others. Pulmonary toxicity, however, represents the most serious adverse reaction limiting the clinical effecacy of this antidysrhythmic drug. Recently, we experienced a case of amiodarone-induced pulmonary toxicity confirmed by open lung biopsy with light and electron-microscopy. So, we report a case of amiodarone-induced pulmonary toxicity with a literature review. PMID- 7495780 TI - Predictive factors for beneficial response to interferon-alfa therapy in chronic hepatitis C. AB - OBJECTIVES: Interferon is the only established treatment for chronic hepatitis C but the host-dependent or virus-related factors affecting the response rate to interferon therapy are not yet clear. The purpose of this study was to investigate the factors predictive of response to interferon-alfa therapy in chronic hepatitis C. METHODS: Twenty-five consecutive patients with chronic hepatitis C were randomized to three regimens of interferon-alfa: group A (n = 7, 3 MU every day for 3 months), group B (n = 8, 3 MU every other day for 3 months) and group C (n = 10, 3 MU every other day for 6 months). We quantified serum HC RNA levels by competitive reverse transcription-polymerase chain reaction (RT PCR) and performed HCV genotyping using type-specific primers deduced form the NS5 region of the HCV genome. We also attempted to identify which demographic, biochemical and histologic factors in addition to virus-related factors would significantly predict beneficial response to interferon by multivariate analysis. RESULTS: Sustained responders were 8 (36.4%), nonsustained responders were 2 (9.1%) and nonresponders were 12 (54.5%) of 22 patients who had received complete therapy. The initial HCV RNA level (logarithmic transformed copy numbers per ml of serum) in sustained responders (5.75 +/- 0.39) was significantly lower than that of nonsustained responders (6.80 +/- 0.71) and nonresponders (6.70 +/- 0.52) (p < 0.05). In multivariate multiple logistic regression analysis, the serum HCV RNA level before therapy was only the independent predictor of a sustained response to interferon-alfa therapy (p = 0.001). CONCLUSIONS: Serum HCV RNA level before therapy was the most useful predictor of a sustained response to interferon-alfa therapy for chronic hepatitis C. PMID- 7495781 TI - HLA-DRB1*0401 is associated with susceptibility to insulin-dependent diabetes mellitus independently of the DQB1 locus. AB - The distribution of DRB1*04 alleles and DRB1/DQB1 haplotypes was analysed in 57 DR4+ caucasoid subjects with insulin-dependent diabetes mellitus (IDDM) and 96 DR4+ healthy controls selected on the basis of DR serology, and the findings were analysed in relation to age at diagnosis of IDDM. DNA samples were amplified using specific DR and DQ primers and hybridized with sequence-specific oligonucleotide probes. A significantly increased combined frequency of DRB1*0401 and 0402 was observed in IDDM subjects aged < or = 12 years at diagnosis (allele frequency 88.4% compared with 62.0% in controls, P < 0.025). There was a non significant increase in DRB1*0401 and 0402 in IDDM subjects < or = 12 years when compared with IDDM subjects > 12 years (P < 0.1). DRB1*0404 was decreased in the total IDDM subject group compared with controls (4.8% vs. 19.0%, P < 0.025) but did not reach statistical significance in the individual age at diagnosis groups. In contrast, the frequency of DQB1*0302 was increased uniformly across both ages at diagnosis groups. In controls DRB1*0401 occurred in haplotype association with DQB1*0301 in a significantly greater frequency than with DQB1*0302. However, 95.0% of DRB1*0401 IDDM subjects were DQB1*0302. DRB1*0404, which was decreased in frequency in IDDM subjects, occurred in association significantly more frequently with DQB1*0302 in controls. These results imply that DRB1 and DQB1 have independent roles as HLA susceptibility genes in IDDM. DQB1 may have a permissive role whereas DRB1 could influence the rate at which underlying disease progresses to clinical IDDM. PMID- 7495783 TI - Immunoprinting excludes many potential susceptibility genes as predisposing to early onset pauciarticular juvenile chronic arthritis except HLA class II and TNF. AB - DNA profiles (immunoprints) were generated for 120 patients suffering from early onset pauciarticular chronic arthritis (EOPA-JCA) and > 500 healthy controls utilizing highly polymorphic microsatellites in the vicinity of immunorelevant genes. Six T cell receptor (TCR) markers for the CD3D, TCRDVAJ, TEA, TCRBV6S1, BV6S3, BV6S7 and BV13S2 genes were analysed. Furthermore markers for the cell surface molecule CD40L, for cytokine genes (IL-1A, IL-2, IFN-alpha, FGF-alpha, TNF-alpha), the chromosomal region of the IRF2 and the cytokine receptor gene IL5RA were studied as well as two polymorphisms within the promotor region of the TNF-alpha gene. Coding region polymorphisms were evidenced indirectly by repeat length variation or they were predicted from the microsatellite distribution profiles and then confirmed by direct sequence analysis. Statistical evaluations were performed with respect to known predispositions, predominance of females (> 80%) and HLA-DR and -DQ haplotypes. Cell surface molecules (TCR, CD40L, IL5RA) as well as almost all cytokines (IL-1A, IFN alpha, FGFA, IRF2 region) were excluded as predisposing in our JCA panel. The TNF-alpha microsatellite alleles (GT)10-12 contribute considerably to manifestation of the disease, in HLA-DRB1*11(12) individuals (RR = 12.8). The TNF-alpha allele is not found in linkage disequilibrium with HLA-DRB1*11(12) and may be present on either chromosome 6. Thus, a novel susceptibility factor probably within the TNFA/TNFB gene region has been identified via linkage with the TNF-alpha microsatellite allele. Apparently complex compositions of the genetic background rather than single genes provide the precondition for manifestation of the autoimmune disease EOPA-JCA. Immunoprinting unravels the variability of the immunological genome via the semi directed microsatellite approach efficiently. PMID- 7495782 TI - Different patterns of HLA-DR antigen expression in normal epithelium, hyperplastic and neoplastic malignant lesions of the breast. AB - Fifteen samples of non-tumoural breast tissue, 24 cases of benign lesions, four biopsies of inflammatory carcinomas and 94 tumour samples of primitive mammary carcinomas were analysed for HLA class II expression. We found, first, that HLA class II antigens were detectable in all cases of non-neoplastic breast tissue. Secondly, HLA class II antigen expression was notably increased in benign neoplasms and hyperplastic lesions. In contrast, only 32 out of 94 carcinomas showed expression of HLA-DR antigens, 17 tumours had HLA-DP antigens and 11 carcinomas were positive for the presence of DQ molecules. The expression of class II antigen was associated with the degree of histological differentiation (P < 0.05) but was independent of stromal leucocytic infiltration. Thirdly, HLA DR was very strongly expressed in intravascular tumoural thrombi, especially in the 'inflammatory carcinomas'. The immunophenotype of inflammatory infiltrate was analysed in benign and malignant lesions. In malignant lesions the mean number of inflammatory cells was significantly higher than in benign lesions. Interestingly, we found no differences in the amount and composition of inflammatory infiltrate between HLA-DR positive and negative tumours. PMID- 7495784 TI - Maternal HLA class II allogenotypes are markers for the predisposition to fetal losses in families of women with unexplained recurrent fetal loss. AB - The HLA allogenotypes DR1/Br, DR3 and DR10 (entitled risk HLA markers) have been reported as being genetic markers for the predisposition to experience unexplained recurrent fetal losses. The aim of the study was to determine whether the putative risk HLA markers might also be markers for the risk of pregnancy loss in sisters and wives of brothers of women with unexplained recurrent fetal losses. Information concerning pregnancy outcomes among the relatives of 146 consecutive women with unexplained recurrent fetal losses was collected. Ninety five of the full sisters, 69 of the full brothers and 50 of the wives of the brothers were HLA typed. Sisters who had experienced at least one previous pregnancy loss (affected women) shared more HLA haplotypes with the proband than unaffected sisters, when the proband was positive for the risk markers (P = 0.02). More affected than unaffected sisters and brothers' wives were positive for the risk markers (P < 0.005 and P < 0.03; respectively). The lowest estimate of the odds ratio for experiencing pregnancy loss among sisters and brothers' wives who were positive compared with those negative for the risk markers was 3.5 (95% credible interval = 1.9-5.8). It is concluded that maternal DR1/Br, DR3 and DR10 allogenotypes seem to be genetic markers for the risk of pregnancy loss among relatives of women with unexplained recurrent fetal losses. The pattern of inheritance suggests a polygenic mode of inheritance with alleles linked to the risk HLA markers interacting with non-HLA linked genes expressed on the fetus or the trophoblast. PMID- 7495785 TI - Nomenclature for factors of the HLA system, 1995. PMID- 7495786 TI - Nomenclature for factors of the HLA System, update June 1995. PMID- 7495787 TI - Organization of P-type ATPases: significance of structural diversity. PMID- 7495788 TI - Antimicrobial peptide pores in membranes detected by neutron in-plane scattering. AB - Antimicrobial peptides isolated from the host defense systems of animals have been shown to exert their activity directly on the lipid bilayer of cell membranes, but the antimicrobial mechanisms are not clear, due chiefly to the difficulty of discerning the high-order structures formed by these peptides in membranes. Previously we have shown that these peptides insert into the membrane when their concentrations exceed a lipid-dependent critical value. With neutron in-plane scattering we now show that inserted alamethicin creates aqueous pores approximately greater than 18 A in diameter. The density of pores is consistent with the assumption that all of the alamethicin is involved in pore formation. Pores were not detected below the critical concentration. Thus concentration dependent pore formation appears to be the molecular mechanism of antimicrobial action. PMID- 7495789 TI - Stereochemistry of chitin hydrolysis by a plant chitinase/lysozyme and X-ray structure of a complex with allosamidin: evidence for substrate assisted catalysis. AB - The plant enzyme hevamine has both chitinase and lysozyme activity. HPLC analysis of the products of the hydrolysis of chitopentaose shows that hevamine acts with retention of the configuration, despite the absence of a nucleophilic or stabilizing carboxylate. To analyze the stabilization of a putative oxocarbonium ion intermediate, the X-ray structure of hevamine complexed with the inhibitor allosamidin was determined at 1.85 A resolution. This structure supports the role of Glu127 as a proton donor. The allosamizoline group binds in the center of the active site, mimicking a reaction intermediate in which a positive charge at C1 is stabilized intramolecularly by the carbonyl oxygen of the N-acetyl group at C2. PMID- 7495790 TI - HO. and DNase I probing of E sigma 70 RNA polymerase--lambda PR promoter open complexes: Mg2+ binding and its structural consequences at the transcription start site. AB - Chemical and enzymatic probing (footprinting) of the reactivity of the promoter DNA backbone is applied to characterize two binary open complexes RPo1 (-Mg2+) and RPo2 (+Mg2+), formed by Escherichia coli RNA polymerase (E sigma 70) at the lambda PR promoter. We report that HO. detects major differences in backbone reactivity between RPo1 and RPo2 in the open region from -4 to +1 relative to the transcription start site. Deoxyribose sugars at positions -4 to +1 of the t (template) strand react with HO. in RPo2 but are relatively protected in RPo1. Binding of Mg2+ to convert RPo1 to RPo2 therefore increases the reactivity of two negatively charged footprinting agents [MnO4-; Suh, W.-C., Ross, W., & Record, M. T., Jr., (1993) Science 259, 358-361; and Fe(EDTA)2-/HO.] at the start site and is required for binding of the negatively-charged initiating nucleotides to the polymerase and the t strand at the start site. We propose that these effects result from binding of two Mg2+ ions to the catalytic carboxyls in the nucleotide binding sites. Except for the key region on the t strand at the start site, the promoter DNA of both RPo1 and RPo2 is continuously protected from DNase I and hydroxyl radical (HO.) cleavage between the -12 and +25 promoter positions. Protection in the upstream region, extending from -13 to about -70, is periodic, with an 11 base pair periodicity indicative of binding of polymerase to a single face of the DNA helix.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7495791 TI - Identification of a "peroxy" intermediate in cytochrome bo3 of Escherichia coli. AB - The respiratory heme-copper oxidases catalyze the reduction of dioxygen to water and link this chemistry to proton translocation. The main subgroups of the enzyme family are the cytochrome c oxidases and the quinol oxidases. For the cytochrome c oxidases, several key intermediates have been described in the oxygen reaction. Two of these (suggested to be "peroxy" and "ferryl" species) are also produced in the reaction of the oxidized enzyme with hydrogen peroxide. However, only a single product (a "ferryl" species) has been reported for the reaction of hydrogen peroxide with the quinol oxidase cytochrome bo3 from Escherichia coli. The same "ferryl" species has also been reported to be produced when two-electron reduced cytochrome bo3 reacts with oxygen, whereas this reaction leads to the "peroxy" intermediate in the cytochrome c oxidases. Consequently, the oxygen reaction has been considered to be different in the two enzyme subgroups. Here we show that both the peroxide reaction and the reaction of the two-electron reduced enzyme with oxygen actually result in primary formation of a hitherto unreported "peroxy" species in cytochrome bo3. This intermediate subsequently relaxes into the "ferryl" species which has been described previously. We conclude that the oxygen reaction is similar in the cytochrome c and quinol oxidases. PMID- 7495792 TI - Interaction of calmodulin with the inducible murine macrophage nitric oxide synthase. AB - The inducible isoform of nitric oxide synthase (iNOS) isolated from murine macrophages copurifies with calmodulin (CaM) as a tightly bound subunit. The exact function of this tightly bound CaM, however, has not been elucidated. In order to probe the function of this unusually strong interaction between iNOS and CaM, a 30 amino acid peptide derived from the putative CaM binding site of iNOS was synthesized. Cross-linking and autoradiographic analyses demonstrated that the peptide and CaM form a 1:1 complex as well as several higher molecular weight complexes. When assayed in the presence of a 12-fold excess of peptide to iNOS, over 90% of the enzymatic activity was inhibited. This inhibition could be prevented with the addition of exogenous bovine CaM to the assay mixture, in a concentration-dependent manner. Native PAGE and Western blot analysis of iNOS treated with peptide revealed the formation of a peptide--CaM complex with CaM derived from iNOS. Moreover, EGTA (5 mM) caused a 30% maximal inhibition of activity that was reversed by the addition of exogenous Ca2+ in a concentration dependent fashion, suggesting a role for Ca2+ in this interaction. EGTA also changed the native PAGE mobility of iNOS and increased the intensity of a band which comigrates with CaM. These results demonstrate that the binding interaction between CaM and iNOS is tight but reversible, requires Ca2+, and is atypical from other known CaM--enzyme interactions. PMID- 7495793 TI - Proton NMR investigation of the [4Fe--4S]1+ cluster environment of nitrogenase iron protein from Azotobacter vinelandii: defining nucleotide-induced conformational changes. AB - This work presents the complete assignment of the isotropically shifted 1H NMR resonances of Azotobacter vinelandii nitrogenase iron protein (Fe protein) to beta-CH2 and alpha-CH protons of the [4Fe--4S]1+ cluster cysteinyl ligands. Four resonances were observed for the reduced Fe protein with chemical shifts of 49, 23, 17, and 13 ppm. T1 measurements and analysis of relative peak areas coupled with one-dimensional nuclear Overhauser effect (NOE) difference spectra were used to assign the two most downfield-shifted resonances (49 and 23 ppm) to cysteinyl ligand beta-CH2 protons and the 17 and 14 ppm resonances to cysteinyl ligand alpha-CH protons. Temperature dependent studies of the isotropically shifted protons revealed both Curie and anti-Curie behavior. These results, along with previous Mossbauer studies of the Fe protein, allowed the assignment of signal A (49 ppm) to four beta-CH2 protons and signal C (17 ppm) to 2 alpha-CH protons of two cysteinyl ligands bound to a mixed-valence iron pair (Fe3(+)--Fe2+) of the [4Fe--4S]1+ cluster. Signal B (23 ppm) was assigned to four beta-CH2 protons, and signal C (17 ppm) and D (13 ppm) were assigned to two alpha-CH protons of two cysteinyl ligands bound to a ferrous pair of irons (2Fe2+). The effects of MgATP, MgADP, and Mg-adenosine-beta, gamma-methylene-5'-triphosphate binding to the Fe protein on the assigned resonances were established and are discussed in the context of nucleotide-induced changes in the protein environment of the [4Fe--4S] cluster.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7495794 TI - Unlike the quaternary structure transition, the tertiary structure change of the 240s loop in allosteric aspartate transcarbamylase requires active site saturation by substrate for completion. AB - The quaternary structural change associated with the homotropic cooperative interactions in Escherichia coli aspartate transcarbamylase (ATCase) is accompanied by various tertiary structural modifications; the most notable one involves the 240s loop formed by residues 230--245 of the catalytic chain. In order to monitor local conformational changes in this region by fluorescence spectroscopy, Tyr-240 has been replaced by a Trp residue, in a mutant enzyme, in which both naturally occurring Trp residues in positions 209 and 284 of the catalytic chains had previously been substituted by Phe residues. This F209F284W240-ATCase still displays homotropic cooperativity for aspartate and undergoes the same T to R quaternary structure change as does the wild-type enzyme. Upon binding of the bisubstrate analogue N-(phosphonoacetyl)-L-aspartate, the fluorescence emission spectrum of this mutant shows a red shift directly proportional to the fraction of catalytic sites occupied by this compound, a maximum value of 4 nm being attained when all six active sites are ligated. An identical shift is observed with the catalytic subunits of this modified enzyme, when all three active sites are occupied. In contrast, the quaternary structural change of the F209F284W240-ATCase, monitored by small-angle X-ray scattering, is complete when only four out of six catalytic sites are occupied. Thus, the 240s loop adopts its final conformation only when the neighboring active site is bound. PMID- 7495795 TI - Use of designed metal-binding sites to study helix proximity in the lactose permease of Escherichia coli. 1. Proximity of helix VII (Asp237 and Asp240) with helices X (Lys319) and XI (Lys358). AB - The lactose permease of Escherichia coli contains two pairs of oppositely charged residues that interact functionally, Asp240 (helix VII)/Lys319 (helix X) and Asp237 (helix VII)/Lys358 (helix XI). Single- and double-His replacement mutants at these positions have been constructed and characterized with respect to transport activity and Mn2+ binding. The following results confirm the functional interactions between both sets of residues: (i) At pH 7.5, where the imidazole is likely to be unprotonated, the double-His mutants Asp237 --> His/Lys358 --> His and Asp240 --> His/Lys319 --> His exhibit significant transport activity while the single-His mutants Lys319 --> His and Lys358 --> His are inactive. (ii) At pH 5.5, where the imidazole is likely to be protonated, the double-His mutants Asp240 --> His/Lys319 --> His and Asp237 --> His/Lys358 --> His are inactive; however, the single-His mutant Lys319 --> His exhibits significant activity. (iii) The single-His mutant Asp237 --> His or ASP240 --> His is inactive at all pH values tested. In addition, a pH titration of Asp237 --> His/Lys358 --> His permease activity exhibits a midpoint at about 6.2. Finally, the purified mutant proteins Asp237 --> His/Lys358 --> His and Asp240 --> His/Lys319 --> His were assayed for Mn2+ binding by electron paramagnetic resonance spectroscopy. Asp237 -> His/Lys358 --> His permease binds Mn2+ with a stoichiometry of unity at pH 7.5, but much less binding is observed at pH 5.5, demonstrating directly that helix VII (Asp237) is in close proximity to helix XI (Lys358).(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7495796 TI - Use of designed metal-binding sites to study helix proximity in the lactose permease of Escherichia coli. 2. Proximity of helix IX (Arg302) with helix X (His322 and Glu325). AB - Engineering divalent metal-binding sites into the lactose permease of Escherichia coli by introducing bis-His residues has been utilized to confirm the proximity of helices VIII (Glu269 --> His) and X (His322) [Jung, K., Voss, J., He, M., Hubbell, W. L., & Kaback, H. R. (1995) Biochemistry 34, 6272] and helices VII (Asp237 --> His) and XI (Lys358 --> His) [He, M. M., Voss, J., Hubbell, W. L., & Kaback, H.R. (1995) Biochemistry 34, 00000--00000]. In this paper, the approach is used to confirm and extend the relationship between helices IX (Arg302) and X (His322 and Glu325) [Jung, K., Jung, H., Wu, J., Prive, G. G., l& Kaback, H. R. (1993) Biochemistry 32, 12273]. Thus, mutants Arg302 --> His, Glu325 --> His, and Arg302 --> His/Glu325 --> His were constructed, and Mn2+ binding was assayed by electron paramagnetic resonance. Mutant Arg302 --> His binds Mn2+ with a KD of about 24 microM and a stoichiometry approximating unity in all likelihood because the His residue at position 302 forms a metal-binding site in conjunction with the native His residue at position 322. Mutant Arg302 --> His/Glu325 --> His also binds Mn2+ with a 1:1 stoichiometry, but the KD is decreased to about 13 microM. The results suggest that Arg302 is sufficiently close to both Glu325 and His322 to form a tridentate metal-binding site in mutant Arg302 --> His/Glu325 --> His. In contrast, replacement of Glu325 with His in permease with a native His residue at position 322 does not lead to Mn2+ binding. The results provide strong support for the helix packing model proposed. PMID- 7495797 TI - Molecular basis for nonadditive mutational effects in Escherichia coli dihydrofolate reductase. AB - Recently, two sets of single, double, and quadruple residue changes within the hydrophobic substrate binding pocket of Escherichia coli dihydrofolate reductase (5,6,7,8-tetrahydrofolate+ oxidoreductase, EC 1.5.1.3) were shown to exhibit nonadditive mutational effects [Huang, Z., Wagner, C. R., & Benkovic, S. J. (1994) Biochemistry 33, 11576--11585]. In particular, the analysis of data for the L28Y, L54F, and L28Y-L54F mutations revealed nonadditive changes in the free energy associated with the substrate and cofactor binding, hydride transfer, and product release steps. Construction of a related set of mutant proteins including L28F and L28F-L54F permits a comparison of similar energy changes and provides a means for assessing differences in the interactions of Phe28 and Tyr28 with both the ligands and the side chains at residue 54. We find a single functional group change, from Phe C4-H to Tyr C4-OH, can influence the additivity of mutational effects and serve as a probe to monitor the appearance of differing enzyme conformations along the reaction pathway through changes in the interaction energy (delta GI). The comparison of additivity/nonadditivity in free energy changes for three interrelated double mutational cycles (WT --> L28F-L54F, WT --> L28Y-L54F, and L28F --> l28Y-L54F) demonstrates that the side chains of positions 28 and 54 interact cooperatively to facilitate hydride transfer by preferentially influencing the enzyme--substrate ground-state complexes. The delta GI data for individual steps also provide evidence for multiple conformations of the enzyme operating during the catalytic cycle. The fact that there are no published examples of the synergistic enhancement of favorable mutational effects is consistent with the expectation that the binding/active site surface of wild-type dihydrofolate reductase has been optimized. PMID- 7495798 TI - Crucial role of an idiosyncratic insertion in the Rossman fold of class 1 aminoacyl-tRNA synthetases: the case of methionyl-tRNA synthetase. AB - A few aminoacyl-tRNA synthetases are characterized by their ability to tightly bind a zinc atom. In the case of Escherichia coli methionyl-tRNA synthetase, a peptide of 21 residues (138--163) having a stable 3-D structure in solution is responsible for zinc binding [Fourmy, D., Meinnel, T., Mechulam, Y., & Blanquet, S. (1993) J. Mol. Biol. 231, 1066--1077; Fourmy, D., Dardel, F., & Blanquet, S. (1993) J. Mol. Biol. 231, 1078--1089]. This peptide, which belongs to a region connecting the two halves of the nucleotide-binding domain of methionyl-tRNA synthetase, is likely to form a modular domain close to the active center of the enzyme. In this study, two residues of the zinc-binding module, Asp138 and Arg139, are shown to contribute to the stabilization of the transition state of the reaction leading to the activation of methionine. Moreover, another residue, Phe135, located at the surface of the zinc-binding domain, is found to possibly guide the tRNA acceptor stem toward the active site of the enzyme during catalysis. The available data indicate an important functional role for the zinc binding module of methionyl-tRNA synthetase, as well as for other modules connecting conserved secondary structure elements in the aminoacyl-tRNA synthetase family. The relation between the occurrence of such variable peptide modules and the expression of both substrate specificity and catalytic efficiency is discussed. PMID- 7495799 TI - Increase of the P1 Lys/Leu substrate preference of carboxypeptidase Y by rational design based on known primary and tertiary structures of serine carboxypeptidases. AB - The P1 substrate preference of serine carboxypeptidases, as expressed by the Lys/Leu ratio, differs by up to 10(5)-fold. Predictions of the major determinants of this preference are made by correlating primary and tertiary structures to substrate preferences. In carboxypeptidase Y from yeast it is predicted that Trp312 constitutes such a determinant, reducing the P1 Lys/Leu substrate preference of this enzyme. The predictions are tested by the construction and kinetic characterization of ten mutant enzymes of carboxypeptidase Y. All of these enzymes exhibit changes in their P1 substrate preference. Generally, small decreases in activity (kcat/Km) are observed with substrates containing uncharged P1 side chains. With substrates containing acidic P1 side chains, i.e., FA-Glu Ala-OH, the activity generally increases slightly, 7-fold in the case of W312K. The most dramatic effects of the Trp312 substitutions are observed with substrates containing basic P1 side chains, i.e., kcat/Km for the hydrolysis of Fa-Lys-Ala-OH with W312E has increased 1150-fold, exclusively as a result of increased kcat values. Similar results have previously been obtained by mutational substitution at position 178 of carboxypeptidase Y. The construction and kinetic characterization of position 178 + 312 double mutants demonstrate that the kinetic effects of substitutions at these two positions are not additive. The P1 Lys/Leu substrate preference of one double mutant, L178D + W312D, has changed 380,000-fold as compared to the wild type enzyme, and the overall P1 substrate preference of this enzyme closely resembles that of carboxypeptidase WII from wheat. PMID- 7495801 TI - Two highly specialized histone H1 proteins are the major chromosomal proteins of the sperm of the sea anemone Urticina (Tealia) crassicornis. AB - The sperm nuclear basic proteins (SNBPs) from a sea anemone (Urticina crassicornis) have been isolated and characterized for the first time. They consist of two sperm-specific members of the histone H1 family with Mr 22,700 and 24,600. They amount to about 60--70% of the total chromosomal sperm proteins. Their amino acid composition and the primary structure of their trypsin-resistant core indicate a strong relation to histone H5 from the nucleated erythrocytes of birds and amphibians as well as to other high sperm-specific H1-like (PL-I) proteins from phylogenetically distant groups. The major presence of histone H1 like protein in the sperm of an organism belonging to such a low phylogenetic group provides experimental support to the hypothesis that SNBPs may all have evolved from a primitive histone precursor. PMID- 7495800 TI - Influence of phosphoenolpyruvate and magnesium ions on the quaternary structure of enzyme I of the phosphotransferase system from gram-positive bacteria. AB - Solution X-ray scattering patterns of enzyme I of the phosphotransferase system from Staphylococcus carnosus indicate an increase in radius of gyration and molecular mass in the presence of Mg2+ or both Mg2+ and phosphoenolpyruvate, indicating a partial dimerization of enzyme I. Mg2+ ions are essential for both the dimerization and the activation, whereas the substrate phosphoenolpyruvate shifts the monomer--dimer equilibrium to the enzymatically active dimer by decreasing the dissociation rate of the phosphorylated dimer. PMID- 7495802 TI - Zinc content and function in human fibroblast collagenase. AB - The zinc contents of samples of human fibroblast collagenase (HFC) purified by different procedures and of samples purified by the same procedure but prepared for analysis by different dialysis protocols have been determined by atomic absorption spectroscopy. Both the purification method and dialysis conditions affect the zinc stoichiometry. Samples purified with and without the use of a zinc-chelate chromatography step and prepared by dialysis against 1 mM CaCl2 had zinc to enzyme ratios of 1.46 and 1.22, respectively. When the first sample was prepared by dialysis against 0 and 10 mM CaCl2, the values changed to 0.15 and 1.94, respectively. Thus, the zinc content of HFC is critically dependent upon the dialysis conditions used to free the enzyme from adventitious metals. This could account for the disparate reports in the literature that give zinc stoichiometries for members of the matrix metalloproteinase (MMP) family of between 1 and 2. The mechanism of inhibition of the one zinc form of HFC by 1,10 phenanthroline (OP) and 4-(2-pyridylazo)resorcinol has been studied in detail. Inhibition by both chelating agents is time dependent and biphasic. There is an initial, instantaneous inhibition characterized by the involvement of a single inhibitor molecule that corresponds to the formation of a ternary complex between the zinc atom, enzyme, and chelator. This is followed by a second, slower phase involving removal of the zinc atom from the enzyme and its chelation by two molecules of inhibitor. Inhibition of four other human MMPs by OP shows similar characteristics and is thought to occur by the same mechanism. PMID- 7495804 TI - Induction of microtubule catastrophe by formation of tubulin-GDP and apotubulin subunits at microtubule ends. AB - The recent discovery that GTP linked to latex beads binds to microtubule ends suggested that nucleotide interactions at this site may play a role in regulating microtubule (MT) dynamics. Evidence for this was sought using DIC microscopy to analyze effects of the free GTP and GDP concentration on the rates of MT elongation and phase transition to rapid shortening (catastrophe, kc). That nucleotide can dissociate and thereby destabilize the plus end by forming nucleotide-free (apotubulin) subunits was indicated by an increase in kc from 0.001 to 0.05 s-1, when the free GTP concentration was reduced from 100 to 0.5 microM, during assembly with 15 microM tubulin--GTP subunits (TuT). That nucleotide can bind to the minus end was indicated by a nearly 5-fold decrease in the rate of elongation when the free GDP concentration was increased from 1.6 to 175 microM, during assembly with a mixture of 36 microM TuT and 54 microM TuD. Further evidence that nucleotide can bind to both ends was provided by the observation that with a mixture of 36 microM TuT and 54 microM TuD, kc was increased from 0.0036 to 0.05 s-1 at the plus end, and from 0.0005 to 0.005 s-1 at the minus end, when the free GDP concentration was increased from 1.6 to 175 microM. Our evidence for destabilization of microtubules by formation of apotubulin and by nucleotide exchange to form terminal TuD subunits suggests that microtubule dynamics can be regulated in cells by an exchange factor that generates apotubulin subunits, or by a GTPase activating protein that forms TuD subunits at microtubule ends. PMID- 7495803 TI - Reconstitution of the endogenous plastoquinone pool in photosystem II (PS II) membrane fragments, inside-out-vesicles, and PS II core complexes from spinach. AB - The possibility of reconstituting a functionally competent endogenous plastoquinone pool in photosystem II (PS II) membrane fragments, inside-out vesicles (ISO-vesicles), and PS II core complexes was analyzed by measuring (i) the characteristic period four oscillation of the oxygen yield due to excitation of dark-adapted samples with a train of short flashes and (ii) laser flash induced transients of the relative quantum yield of chlorophyll fluorescence. The data obtained revealed that (a) an endogenous pool capacity comparable to that of intact thylakoids can be restored in PS II membrane fragments and ISO-vesicles by a sonication treatment using native plastoquinone-9, (b) a more pronounced oxygen oscillation pattern arises in PS II core complexes after application of the same reconstitution procedure, (c) the extent of the endogenous pool restoration at a ratio of 15 quinone molecules per PS II in the reconstitution assay strongly depends on the nature of the quinone molecule [maximum effects can be only achieved with PQ-9, while at the same concentration ubiquinone-45 (UQ-9) is almost inefficient], and (d) a sonication step is required for stable insertion of PQ-9 into PS II preparations. Measurements of the reconstruction degree as a function of the structure of different quinones with selected properties lead to the conclusion that specific binding domains exist in PS II in addition to the QB site. These domains exhibit a surprisingly high specificity for the type of quinone that can be bound. On the basis of a comparison of the results obtained, the structure of the quinone head group seems to be more important than the large hydrophobic side chain and/or the general lipophilicity of the compound. PMID- 7495805 TI - Pathway of proton transfer in bacterial reaction centers: further investigations on the role of Ser-L223 studied by site-directed mutagenesis. AB - The role of Ser-L223 in proton transfer to reduced QB in the reaction center (RC) from Rhodobacter sphaeroides was studied by site-directed replacement of Ser with residues having different proton donor properties, e.g., the aliphatic residues Ala and Gly, the hydroxyl residue Thr, the amide residue Asn, the sulhydryl residue Cys, the imidazole residue His, and the carboxylic acid residue Asp. Compared to native reaction centers, RCs with Ala or Asn at L223 had greatly reduced (approximately 300-fold) proton-coupled electron transfer rates, kAB(2), associated with the second electron reduction of QB (QA(-)QB(-) + H+ --> QAQBH-). In contrast, RCs containing Thr, Asp, or Gly at L223 retained fast proton-coupled electron transfer rates. RCs with His or Cys at L223 did not bind the secondary quinone QB. These results show that kAB(2) is larger when a good proton transfer group, e.g., a hydroxyl residue (Ser, Thr) or a carboxylic acid (Asp), occupies the L223 site, supporting the proposal that Ser-L223 is a component of a proton transfer chain [Paddock, M. L., McPherson, P. H., Feher, G., & Okamura, M. Y. (1990) Proc. Natl. Acad. Sci. U.S.A. 87, 6803--6807]. The surprising result that kAB(2) is not significantly reduced in RCs with Gly at L223 suggests that a water molecule functionally replaces the missing Ser hydroxyl group in the mutant RCs. The importance of Ser-L223 in internal proton transfer reactions within the RC is discussed. PMID- 7495806 TI - Interaction of bovine brain tubulin with the 4(1H)-pyrizinone derivative IKP104, an antimitotic drug with a complex set of effects on the conformational stability of the tubulin molecule. AB - The ligands of tubulin have proved to be excellent probes for the conformation of the tubulin molecule. The most varied in their effects on tubulin are those ligands which are competitive or noncompetitive inhibitors of vinblastine binding. The 4(H)-pyrizinone derivative 2-(4-fluorophenyl)-1-(2-chloro- 3,5 dimethoxyphenyl)-3-methyl-6-phenyl-4(1H)-pyridinone [sequence: see text] (IKP104) is a novel antimitotic drug which inhibits microtubule assembly in vitro and in vivo and polymerizes tubulin into spiral filaments. Using a fluorescence assay, we found that IKP104 appears to bind to tubulin at two classes of site, differing in affinity. IKP104 also blocks formation of an intrachain cross-link in beta tubulin, induced by N,N"-ethylenebis(iodoacetamide), linking Cys12 to either Cys201 or Cys211. IKP104 appears to belong to that group of tubulin ligands which includes vinblastine, maytansine, rhizoxin, phomopsin A, dolastatin 10, and halichondrin B. An unusual effect of IKP104 is that it greatly enhances the decay or apparent unfolding or opening of the tubulin molecule. The sulfhydryl titer of tubulin is doubled and the exposure of hydrophobic areas on the tubulin molecule is tripled by IKP104. These effects of IKP104 are counteracted by vinblastine, maytansine, and phomopsin A, suggesting that IKP104 may be competing with these other drugs for binding to tubulin. However, the effects are also counteracted by colchicine and podophyllotoxin, implying a more complex effect, namely, that IKP104 and colchicine, even when both are bound to tubulin, are competing for their effects on the same domain of tubulin. Surprisingly, when IKP104 is used in conjunction with colchicine, binding of colchicine to tubulin is strongly stabilized.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7495807 TI - Activation of human immunodeficiency virus gene expression by ultraviolet light in stably transfected human cells does not require the enhancer element. AB - Ultraviolet light (UV) exposure of cells infected with human immunodeficiency virus type I (HIV) or transfected with HIV reporter genes increases virus directed gene expression. Here we report the mapping of the UV response on the long terminal repeat (LTR) by using human cells stably transfected with HIV promoter plasmids harboring different mutations and controlling the expression of the chloramphenicol acetyltransferase (cat) reporter gene. Promoter mutation analysis revealed that no specific upstream region of the LTR was associated with UV activation, although a significant decrease was observed with mutations in the basal promoter elements Spl and TATA. Most importantly, UV activation was not diminished by removal of the - 119 to -69 region encompassing the LTR enhancer region or, more specifically, by point mutations in the NF -kappa B binding elements. Consistent with this result, we found that the phorbol ester (PMA) response, which is known to act through the enhancer, occurred independently and was synergistic with the UV response. Removal of the -119 to -69 region did not affect UV activation; however, it resulted in total abrogation of the PMA response. These results suggest that UV activation is distinct from NF -kappa B activation and does not act through the enhancer in stably transfected cells. This is in dramatic contrast to what is found with transient expression analysis of these responses. Lastly, RNA protection experiments revealed that UV may act on preassembled basal transcription complexes by allowing elongation of nascent short mRNAs generated from the LTR.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7495808 TI - Ionizing radiation activates nuclear factor kappa B but fails to produce an increase in human immunodeficiency virus gene expression in stably transfected human cells. AB - We have investigated the differential effects of ultraviolet light(UV) and ionizing radiation (IR) on human immunodeficiency virus type 1 (HIV) and c-jun expression in HIVcat/HeLa cells. This cell line harbors integrated copies of the chloramphenicol acetyltransferase (cat) gene under control of the HIV promoter. Both UV and IR increased the binding of nuclear proteins to an oligonucleotide spanning the HIV enhancer region nuclear factor kappa B sites, but only UV increased HIVcat steady-state mRNA and CAT activity. By comparison, transcription of the cellular c-jun gene increased after both types of radiation, but UV was at least 5-fold more effective than IR despite the fact that protein binding to an activator protein 1 oligonucleotide increased similarly after both UV and IR. The lack of HIVcat transcriptional response after IR does not appear to be the result of the repressor binding to upstream promoter elements since cells stably transfected with different HIV promoter deletions showed a lack of response to IR distinguishable from that of the intact promoter. While our findings indicate no correlation between increased binding of transcription factors to upstream promoter elements and increased expression of these genes after radiation, we did observe major differences in how UV and IR affected chromatin structure. UV produced extensive global chromatin decondensation, whereas IR did not, as seen in the microscope and determined by the increased susceptibility of chromatin to micrococcal nuclease digestion.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7495809 TI - Analysis of the proteolytic activity of a recombinant form of apolipoprotein(a). AB - We have analyzed the proteolytic activity of a recombinant form of apolipoprotein(a) [r-apo-(a)]. A mutant 17-kringle from of r-apo(a) was engineered that contained a serine to arginine substitution which reinstates the tissue-type plasminogen activator (tPA) as determined by SDS-PAGE and fluorography and by Western blot analysis. However, tPA cleavage did not result in an active protease as both wildtype r-apo(a) and the mutant, either free or incorporated into r-Lp(a) particles, were uniformly inactive against a variety of chromogenic serine protease tripeptide substrates. To assess whether the large number of kringle IV repeats present in apo(a) inhibits proteolytic activity, we generated truncated forms of the Ser-->Arg proteolytic activity, we generated truncated forms of the Ser-->Arg mutant containing one or 10 kringle IV repeats. These truncated versions of r-apo(a) were susceptible to cleavage by tPA but were inactive against the plasmin substrate S-2251. Treatment of the Ser-->Arg mutant of the 17-kringle r-apo(a) with tPA and diisopropylflurophosphate (DFP) did not result in modification of the mutant protease domain by DFP. Finally, we incubated r-apo(a) or r-Lp(a) particles formed in vitro with purified human LDL; no degradation of LDL was observed after 16 h at 37 degrees C. The results of this study suggest that one or more of the substitutions present in the protease domain of apo(a), in addition to the Arg-->Ser substitution, render apo(a) proteolytically inactive. PMID- 7495810 TI - Catalytic properties of hairpin ribozymes derived from Chicory yellow mottle virus and arabis mosaic virus satellite RNAs. AB - Regions of the negative strands of the satellite RNAs of chicory yellow mottle virus (sCYMV1) and arabis mosaic virus (sArMV) have similarity in sequence and predicted secondary structure compared to the tobacco ringspot virus satellite RNA (sTRSV) hairpin ribozyme, suggesting that they may also be catalytic RNAs of a similar type. Our experiments show that the hairpin ribozyme-like sequences derived from sCYMV1 and sArMV have high phosphodiesterase activity. The Kcat values determined are similar to that of the highly active native sTRSV hairpin ribozyme under the same conditions, although the Km values are much higher. The Km of the sArMV ribozyme was reduced 3-fold, with no change in kcat, by extending substrate hybridization in helix 2. Additionally, the three hairpin ribozymes prefer different GUX sequences on the immediate 3'-side of the cleavage site. The sTRSV hairpin ribozyme cleaves GUX substrates with catalytic efficiencies in the relative order GUC >> GUU > GUG = GUA. The sCYMV1 ribozyme cleaves GUA > GUC, GUG, GUU. The sArMV ribozyme prefers GUA > GUG > GUU > GUC. The functional domain, regulating substrate selection at this position, must reside in the nucleotides that vary between the ribozyme--substrate complexes. The sTRSV ribozyme is most efficient at cleaving GUC complexes, while the sCYMV1 and sArMV ribozymes are most efficient for cleaving GUA-containing sequences. PMID- 7495811 TI - DNA- and chromatin-condensing properties of rat testes H1a and H1t compared to those of rat liver H1bdec; H1t is a poor condenser of chromatin. AB - Histones H1a and H1t are two major linker histone variants present at the pachytene interval of mammalian spermatogenesis. The DNA- and chromatin condensing properties of these two variants isolated from rat testes were studied and compared with those from rat liver. For this purpose, the histone H1 subtypes were purified from the respective tissues using both acid and salt extraction procedures. Circular dichroism studies revealed that acid exposure during isolation affects the alpha-helical structure of both the globular domain (in the presence of 1 M NaCl) and the C-terminal lambda-tail (in the presence of 60% trifluoroethanol). The condensation of rat oligonucleosomal DNA, as measured by circular dichroism spectroscopy, by the salt-extracted histone H1 was at least 10 times more efficient than condensation by the acid-extracted histone H1. A site size of 16-20 base pairs was calculated for the salt-extracted histone H1. Among the different histone H1 subtypes, somatic histone H1bdec had the highest DNA condensing property, followed by histone H1a and histone H1t. All the salt extracted histones condensed rat oligonucleosomal DNA more efficiently than linear pBR-322 DNA. Histones H1bdec and H1a condensed histone H1-depleted chromatin, prepared from rat liver nuclei, with relatively equal efficiency. On the other hand, there was no condensation of histone H1-depleted chromatin with the testes specific histone H1t. A comparison of the amino acid sequences of histone H1d (rat) and histone H1t (rat) revealed several interesting differences in the occurrence of DNA-binding motifs at the C-terminus. A striking observation is the presence of a direct repeat of an octapeptide motif K(A)T(S)PKKA(S)K(T)K(A) in histone H1d that is absent in histone H1t. PMID- 7495812 TI - Interaction between the Escherichia coli Regulatory protein TyrR and DNA: a fluorescence footprinting study. AB - The Escherichia coli regulatory protein TyrR controls the expression of eight transcription units that encode proteins involved in the biosynthesis and transport of aromatic amino acids. It is a homodimer of 57 600 subunit molecular weight and has a binding site for ATP and weak ATPase activity. In the presence of ATP, TyrR binds tyrosine, which induces self-association of TyrR from a dimer to a hexamer. This report examines the interaction of TyrR with a 42 bp DNA oligonucleotide containing a centrally located binding site for TyrR (TyrR box). Replacement of a thymidine residue with an aminouridine residue at positions 7, 9, 13, 15, 19, 22, and 26 from one end of the 42mer enables labeling with fluorescein and successive placement of the label along the major groove of the DNA. The fluorescence footprinting of the oligonucleotide was followed using steady-state and time-resolved fluorescence methods. Binding of the TyrR dimer caused significant changes in the fluorescent properties of the labels attached to positions 13, 15, and 26, suggesting the involvement of these bases in the binding of the protein. Except for the position 15 conjugate, binding of the TyrR dimer caused little change in fluorescence intensity. Therefore, fluorescence anisotropy was used to follow the binding equilibrium. The fluorescence of the position 15 conjugate increased 1.6-fold on binding TyrR, suggesting that the fluorophore was in close contact with the protein. For all conjugates, the addition of tyrosine at the end of the titration with TyrR increased the anisotropy markedly, suggesting that the hexameric form of TyrR could bind the oligonucleotide. Two rotational correlation times were found for the labeled conjugates: one reflecting the motion of the probe at its point of attachment to the DNA (220-290 ps), the other reflecting the global tumbling of the labeled oligonucleotide (14-21 ns). On binding TyrR, changes in the correlation times and their associated amplitudes and changes in the range of angular motion of the probe depended on the position of the label. Evidence is presented that the binding of the TyrR hexamer, but not the TyrR dimer, affects regions that flank the binding sequence. The results support the hypothesis that the binding of the TyrR hexamer is responsible for interaction between tandem TyrR boxes in the tyrR regulon. PMID- 7495813 TI - Kinetics and thermodynamics of intermolecular catalysis by hairpin ribozymes. AB - The hairpin ribozyme, derived from the negative strand of the satellite RNA of tobacco ringspot virus, belongs to the class of small catalytic RNAs that cleave RNA to generate 2',3'-cyclic phosphate and 5'hydroxyl termini and ligate these termini in the reverse reaction to form 3',5'-phosphodiesters. Rate and equilibrium constants for binding, dissociation, cleavage, and ligation steps in the kinetic mechanism were determined using a series of hairpin ribozyme/substrate pairs that differed in the sequence and length of the intermolecular base-paired helices. All hairpin variants cleaved with rate constants of approximately 0.3 min-1 at pH 7.5 in 10 nM MgCl2 at 25 degrees C, regardless of the length or sequence of the intermolecular helices. A rate constant of approximately 3 min-1 was determined for an intermolecular ligation reaction in which both cleavage products were supplied to the ribozyme in trans. Thus, the hairpin favored ligation over cleavage by 10-fold when the ribozyme was saturated with cleavage products. Binding rate constants for cleavage substrates and products were comparable to values reported for other catalytic RNAs but were somewhat slower than binding rates typical of small RNA helices. Substrate dissociation rate constants were much slower than cleavage rate constants for all substrates. Because virtually every substrate that was bound was cleaved before it could dissociate, KMS values were not the same as KdS values. Instead, KMS reflected the ratio of cleavage and substrate binding rate constants and had the same value of approximately 30 nM for all substrates. Calculations based on empirically determined free energy parameters for simple RNA helices indicated that complexes between ribozymes and 5'-cleavage products were slightly less stable than simple helices with the same sequences. In contrast, affinities between ribozymes and cleavage substrates and between ribozymes and 3'-cleavage products were stronger than expected for simple duplexes by about -2.5 kcal/mol, evidence of stabilizing interactions in addition to those contributed by helical base pairs. This kinetic and thermodynamic study demonstrates that the kinetic mechanism of the hairpin ribozyme is distinct from the kinetic mechanisms of other well-characterized ribozymes and provides a foundation for further exploration of the hairpin structure and catalytic mechanism. PMID- 7495814 TI - Binding of phenylarsenoxide to Arg-tRNA protein transferase is independent of vicinal thiols. AB - Reversible enzyme inhibition by phenylarsenoxides is generally taken to indicate the presence of functionally important vicinal thiol groups. Arginyl aminoacyl tRNA transferase from eukaryotes is potently inhibited by phenylarsenoxides and possesses one or more essential sulfhydryl groups (Li, J., & Pickart C. M. (1995) Biochemistry 34, 139-147]. To map the putative Cys residues that mediate arsenoxide binding to the transferase from Saccharomyces cerevisiae, we systematically mutagenized the 15 Cys residues of the transferase, singly and in combination, to Ala (13 Cys) or Ser (2 Cys). Six mutant enzymes, encompassing all 15 Cys residues of the transferase, were characterized in detail. The results revealed that Cys-20, Cys-23, and Cys-94 and/or Cys-95 were important for activity, since mutations at these positions reduced activity by 100-fold (Cys-94 and Cys-95 were mutated simultaneously). Surprisingly, however, all of the mutant enzymes retained the ability to bind a radioiodinated phenylarsenoxide derivative, with undiminished stoichiometry and affinity. All of the mutant enzymes also remained susceptible to irreversible reaction with a bifunctional phenylarsenoxide bearing a paraalkyl halide substituent. Prior reaction of the enzyme with the bifunctional reagent blocked subsequent binding of the radiolabeled phenylarsenoxide, indicating that these two reagents bind at a single common site. These results indicate that high-affinity binding of trivalent arsenicals can occur by a thiol-independent mechanism. PMID- 7495815 TI - Methylation of minimalist 23S rRNA sequences in vitro by ErmSF (TlrA) N methyltransferase. AB - ermSF (synonym tlrA) from Streptomyces fradiae NRRL 2702 confers resistance to the macrolide-lincosamide- streptogramin type B (MLS) superfamily of antibiotics. ErmSF specifically methylates Bacillus subtilis 23S rRNA in vitro at A2085 (B. subtilis coordinate, which is equivalent to the Escherichia coli coordinate A2058). In the present studies, partial B. subtilis 23S rRNA sequences containing portions of the peptidyltransferase circle which include A2085 were constructed in order to identify structural requirements needed for RNA to function as substrate of ErmSF. A model methylase substrate based on the 41-nucleotide construct DK111, ggCCUAUCCGUCGCGGGUUCGCCCGCGACAGGACGGA*AAGA, had methyl-acceptor activity. This sequence contains 23S rRNA stem 73 [Stade, K., et al. (1994) Nucleic Acids Res. 22, 1394-1399] underlined, flanking a tetraloop-like (UUCG), and the impaired sequence AAAGA, at the 3' end containing A2085 (A*). A set of systematic alterations introduced into the sequence suggested that the four unpaired nucleotides in stem 73 are necessary for methyl-acceptor activity, whereas inversion of 11 out 13 paired bases in stem 73 conferred no significant reduction in methyl-acceptor activity. PMID- 7495816 TI - Roles of oxygen and photoinduced acidification in the light-dependent antiviral activity of hypocrellin A. AB - Hypocrellin A displays photoinduced antiviral activity, in particular against the human immunodeficiency virus (HIV), as does its counterpart, hypericin. Although hypocrellin A, like hypericin, executes an excited-state intramolecular proton transfer, it differs from hypericin in two important ways. Unlike hypericin, hypocrellin A absolutely requires oxygen for its antiviral activity. Also, whereas we have previously demonstrated that hypericin functions as a light induced proton source, we do not observe that hypocrellin A acidifies its surrounding medium in the presence of light. These results are discussed in the context of the ground- and excited-state photophysics of hypericin and its mechanisms of photoinduced virucidal activity. PMID- 7495817 TI - Functional reconstitution of the large-conductance, calcium-activated potassium channel purified from bovine aortic smooth muscle. AB - The charybdotoxin (ChTX) receptor has been purified from bovine aortic smooth muscle using conventional chromatographic techniques and sucrose gradient centrifugation. Fractions from the final sucrose gradient purification were enriched in specific binding of monoiodinated ChTX (125i-ChTX) approximately 2000 fold over native sarcolemmal membranes. The ChTX binding activity correlated with the presence of two polypeptides of 65 (alpha) and 31 (beta) kDa. Using the cross linking reagent, disuccinimidyl suberate, 125I-ChTX was specifically incorporated into a polypeptide of approximately 31 kDa. Cross-linking and binding of 125I ChTX to the purified ChTX receptor was inhibited by ChTX, iberiotoxin (IbTX), and tetraethylammonium (TEA). Liposomes containing the purified ChTX receptor were incorporated into planar lipid bilayers. In symmetric 150 mM KC1, the channels observed were > 20-fold more selective for potassium over sodium and exhibited a large, single-channel conductance of 323 +/- 2.5 pS in charged lipids and 249 +/- 7 pS in neutral lipids. Depolarizing membrane potentials increased the open probability of the purified channels e-fold per 11.5 +/- 0.3 mV, while intracellular calcium increased the open probability according to a third power (2.9 +/- 0.2) relationship. Mean channel closed durations decreased while open times slightly increased as membrane potential and calcium concentration were elevated. The distributions of open and closed durations were well described by the sums of three and five to six exponential components, respectively. Purified maxi-K channels were blocked with micromolar affinity by external TEA and with nanomolar affinity by extracellular IbTX and ChTX. Kinetics of ChTX block of the purified channel revealed an equilibrium dissociation constant for toxin block 4.6 +/- 0.7 nM under conditions of physiological ionic strength. The purified maxi-K channel displays many of the biophysical and pharmacological properties of maxi-K channels derived from native tissue. PMID- 7495818 TI - Identification and biochemical characterization of an 80 kilodalton GTP binding/transglutaminase from rabbit liver nuclei. AB - The primary aim of these studies was to identify and biochemically characterize GTP-binding proteins in the nucleus. We found that an 80 kDa protein was responsible for the majority of the GTP-binding activity detected in rabbit liver nuclear preparations as assayed by photoaffinity labeling with [alpha-32P]GTP. The GTP-binding activity was partially extracted only after treatment of nuclear envelope preparations with 0.5 M NaCl and 1% Triton-X 100, which suggested that this GTP-binding protein was a component of the nuclear pore/lamina fraction. The Triton-X-100/NaCl-solubilized 80 kDa protein was purified by a series of steps that included DEAE-Sephacel, Mono-Q, and Ultrogel AcA34 chromatographies. Microsequence analysis of two peptides generated by trypsin digestion of the 80 kDa protein indicates that it shares sequence similarity with the tissue transglutaminases. Purified preparations of the 80 kDa protein show a Ca(2+) stimulated transglutaminase activity, as assayed by the incorporation of [3H]putrescine into caesin, which is strongly inhibited by GTP but not by GDP. A 36 kDa GTP-binding protein copurified with the 80 kDa GTP-binding protein through all of the chromatography steps and sequence analysis suggests that the 36 kDa protein represents a proteolytic fragment of the amino-terminal half of the 80 kDa protein and thus serves to mark the GTP-binding domain within the 80 kDa protein. The 36 kDa fragment has a significantly higher efficiency of [alpha 32P]GTP incorporation compared to the 80 kDa protein, suggesting that the carboxyl-terminal half of the GTP-binding protein/transglutaminase imparts a negative constraint on GTP-binding activity or on the subsequent incorporation of radiolabeled GTP.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7495819 TI - The contribution of the conserved hinge region residues of alpha1-antitrypsin to its reaction with elastase. AB - The hinge region of serpins is a conserved sequence of 8 amino acids located 7 residues away from the scissile bond at P8 to P15, on the edge of the protease binding domain. In the inhibitory serpins the P8 to P12 residues of this motif are usually small side-chain amino acids, most commonly alanine. Each of these residues in alpha1-antitrypsin was mutated to a glutamate, and the effect of a hinge-region glutamic acid substitution was found. While substitutions at positions P10 and P12 affected the inhibitory characteristics of alpha1 antitrypsin, substitutions at positions P7, P8, P9, and P11 had no effect on inhibition. Thus, the conservation of residues with small side chains at the latter positions does not appear to be related to an essential function in the inhibitory mechanism. Following the glutamate substitution at P10, alpha1 antitrypsin remained a rapid inhibitor of elastase, but the elastase--serpin complex slowly broke down to yield active elastase and cleaved alpha1 antitrypsin. The glutamate substitution at P12 caused the resultant molecule (P12 Ala-->Glu) to become a partial substrate of elastase such that four moles of inhibitor were required to inhibit one mole of enzyme, and led to a 12-fold decrease in the association rate constant. The data could be interpreted in terms of the suicide substrate inhibition model for serpin-protease interactions and allowed a further refinement of the role of the hinge region in this process. PMID- 7495820 TI - Deletion of 343 amino acids from the carboxyl terminus of the beta-subunit of the insulin receptor inhibits insulin signaling. PMID- 7495822 TI - Coping, social support, and depressive symptoms in adolescent mothers. AB - PURPOSE: The purpose of this study was to determine how the degree of depressive symptomatology among adolescent mothers differentiated maternal outcomes, social support, and coping at 6 months postpartum. METHODS: Fifty primiparous adolescents, predominantly black and of low income, participated in the study. During a home visit at 6 months postpartum, a research assistant, blind to the study's hypotheses, administered questionnaires related to depressive symptoms, coping with motherhood, social support, maternal confidence, and maternal gratification. Feeding and teaching interactions between mother and child were observed. RESULTS: Fifty-six percent of the subjects had no depressive symptoms (group 1), 20% reported mild symptoms (group 2), and 24% reported moderate to severe symptoms (group 3). Adolescents with mild or moderate depressive symptoms had more negative feeding interactions with their infants, reported less maternal confidence and gratification, and used more emotion-focused coping than their nondepressed counterparts. While the groups did not differ in the frequency with which they received social support, adolescents with mild or moderate depressive symptoms were less satisfied with this support. No differences were found among groups with regard to the size of their conflicted networks. CONCLUSIONS: This study extends the research findings related to depressive symptoms among adult mothers to a sample of adolescent mothers. Although the study design is crosssectional and therefore causal sequences cannot be determined, the results identify factors that may place certain adolescents at risk for problems in parenting. The authors suggest screening adolescent mothers during the first postpartum year for symptoms of depression. PMID- 7495821 TI - Factors affecting pediatric residents' intentions to screen for high risk behaviors. AB - PURPOSE: To determine the factors associated with the intentions of pediatric residents to: 1) screen adolescents for high risk behaviors using the Guidelines for Adolescent Preventive Services (GAPS) and 2) to record the results in the medical record. METHODS: 64 pediatric residents at a university children's hospital were given seven scenarios and were asked to rate on a five point Likert scale (1 = always, 5 = never) the likelihood that they would ask and document adolescent patient responses about sexual activity/birth control, alcohol/drug use, depression/suicidal ideation, fighting/coping with anger, and nutrition/eating disorders. The association of patient-related factors (e.g., gender, presenting symptom, medical setting, known private provider, and presence of a chronic illness) and resident-related factors (e.g. gender, post graduate level, future plans, and a prior adolescent rotation) on intentions to screen was determined using analysis of variance for ranked data. RESULTS: Residents indicated they were most likely to ask adolescents about sexual activity (mean Likert = 2.6, median Likert = 2, range 1-5) (P=0.014). Residents were more likely to screen for substance use (2.8, 3, 1-5) than nutritional issues (3.5, 4, 1-5) (P < or = 0.0001), and were least likely to ask about depression (3.8, 4, 1-5) and fighting (3.9, 4, 1-5) (P < or = 0.0001). Although residents reported that they were likely to record screening results in the medical record (median Likert score = 1), they noted the most common reasons for not recording to be fear that parents would see the record, lack of time, and the feeling that negative responses to screening need not be recorded. The patient-related factors that significantly affected residents' intentions to screen included: presenting symptom, medical setting, and the presence of a chronic illness (P < or = 0.05). Female gender of the patient increased only the likelihood of nutrition screening. Male residents reported a greater intention to screen for depression (P = 0.046). Post-graduate level and a prior adolescent rotation influenced the intention to screen for most behaviors. CONCLUSIONS: This study found that residents are more likely to indicate that they would screen for sexual activity and substance use than for nutritional disorders, depression, and fighting. Several patient-related and resident-related factors influence the likelihood of residents to screen for various high risk behaviors. Educational efforts are needed to increase residents' screening for high risk behaviors. PMID- 7495823 TI - Iguana-associated salmonellosis in a young adult. AB - PURPOSE: Review a case of Salmonella infection in a young adult related to handling of an infected iguana. METHODS: Case report. RESULTS: Most cases of Salmonella infection related to handling of reptiles have occurred in children. We report a case of Salmonella diarrhea in a 19-year-old male who kept a pet iguana. The iguana was asymptomatic, but Salmonella grew from stool specimens. CONCLUSIONS: Those who keep iguanas as pets, which are particularly attractive to adolescents and young adults, should be aware that iguanas frequently carry Salmonella. Those caring for adolescents and young adults should always inquire into the pet-keeping habits of their patients when illnesses develop. PMID- 7495824 TI - Is the clinical spectrum of gonorrhea changing? AB - PURPOSE: The study's objective was to determine whether the high rates of gonorrhea in one inner city teen health care program were due to an increase in detection of asymptomatic cases and consequently a decrease in symptomatic gonococcal infections over a 4 1/2 year study period. METHODS: A chart review was conducted of all females ages 11-19 years with gonorrhea during the last six months of 1988 and all of 1990 and 1992. The presenting symptoms, physical signs, diagnosis, co-infections, and history of STD contacts were analyzed. RESULTS: Four hundred and twenty-five charts were reviewed: 104 (24%) cases in the 11-15 year old age group, 169 (40%) in the 16-17 year old age group and 152 (36%) in the 18-19 year old age group. Approximately two-thirds had physical signs consistent with a sexually transmitted disease. About 25% presented as a contact of someone with a known STD or with symptoms suggestive of a STD. Fewer than 2% had pelvic inflammatory disease. The majority of all groups had symptoms, though older teenagers provided complaints significantly more often (p < 0.01). Teenagers under 15 years that were significantly more likely to have no symptoms or signs (p < .05). CONCLUSIONS: There were no significant changes detected in the percent of symptomatic subjects over the 4 1/2 year period, i.e., the clinical spectrum of gonorrhea remained unchanged. PMID- 7495825 TI - Female circumcision: a report of four adolescents. PMID- 7495826 TI - Predicting resting blood pressure at eighteen years: the Dunedin Study. AB - PURPOSE: To examine the value of repeated blood pressure screening during childhood, and exercise testing in adolescence, for predicting resting blood pressure at eighteen years. METHOD: Regular measures of blood pressure and other variables in a birth cohort from 7 to 18 years. Cardiovascular responses to exercise testing at 15 and 18 years. RESULTS: The best predictors of resting blood pressure at 18 years were body size at 18 years and resting blood pressure at 15 years. Neither exercise and recovery blood pressures at 15 years, nor the way blood pressure had tracked through childhood, added importantly to the prediction. CONCLUSION: We question the value of screening for hypertension during childhood, either by regular resting measures or by exercise testing in adolescence. PMID- 7495827 TI - A medico-social profile of adolescent rag pickers handling hospital wastes. PMID- 7495829 TI - Select school-based health center publications. PMID- 7495828 TI - State initiatives to support school-based health centers: a national survey. PMID- 7495830 TI - Sexually-transmitted disease in female adolescents: effects of psychosocial factors and high risk behaviors. AB - BACKGROUND: Although previous studies have reported behavioral correlates of sexually-transmitted diseases (STDs) and psychosocial correlates of risky behavior, research has not examined the linkages between psychosocial attributes, behavioral patterns, and actual STD acquisition within the same adolescent sample. We examined the effects of five psychosocial factors on risky sexual behavior and substance use, and their direct and indirect effects on STD acquisition. METHODS: A multiethnic sample of 571 sexually-active female adolescents (ages 13-19 years) was recruited from family planning clinics. Subjects completed questionnaires prior to their gynecologic examination and STD testing. RESULTS: A multiethnic sample of 571 sexually-active female adolescents (ages 13-19 years) was recruited from family planning clinics. Subjects completed questionnaires prior to their gynecologic examination and STD testing. RESULTS: Female adolescents who were heavy substance users, who felt little control over their sexual behavior, and had a greater number of friends who engaged in risky sexual behavior were themselves more likely to engage in risky sexual behavior. Risky sexual behavior was associated with positive STD status. Subjects who used psychoactive substances during sex reported higher numbers of sexual partners and were more likely to have an STD. CONCLUSIONS: This research has identified possible causal paths by which psychosocial factors may influence specific behaviors that are predictive of STDs in adolescent females. PMID- 7495831 TI - The effectiveness of selected risk factors in mediating gender differences in drinking and it problems. AB - PURPOSE: The purpose of this study was to investigate gender differences in adolescents' alcohol use, adverse consequences of alcohol and other drug (AOD) use and the effectiveness of selected risk factors in mediating gender differences. METHODS: A cross-sectional survey design was adopted. Data were collected from a sample of high school students (n=919) in one midwestern community. Four criteria for examining potential mediated relationships were addressed. RESULTS: Expected differences in relation to males' higher levels of frequent and heavy drinking, adverse consequences of AOD use and risk factors were found. All risk factors were significantly related to adverse consequences, but alienation was not related to drinking behavior. In a one-way ANOVA, the main effects of gender were significant for both drinking behavior and adverse consequences of AOD use. In a two-way ANOVA, the main effects of a risk factor index reduced the main effects of gender substantially. CONCLUSIONS: Controlling risk factor levels mediates gender differences in the outcomes, drinking behavior and adverse consequences of AOD use. PMID- 7495832 TI - The associations between immigrant status and risk-behavior patterns in Latino adolescents. AB - PURPOSE: The purpose of this study was to identify differences in patterns of risk-taking behavior among Latino adolescents with respect to immigrant status and in comparison to their native non-Hispanic white counterparts. METHODS: In fall of 1988 and spring of 1989 a confidential Teen Health Risk Survey was administered to 1,789 students at two high schools in northern California, both of which have a school health center on-site, and which had large numbers of Latino students enrolled. We divided the Latino adolescent population into two groups: Latinos who were born in the United States (native-born Latinos) and Latinos who had immigrated to the United States (Latino immigrants). Eight different risk-taking behaviors were identified for this study: alcohol, cigarette, marijuana, illicit drug use, self-violence, drunk driving, unintended pregnancy, and violence. RESULTS: We found that in general, Latino students engaged in a greater number of risk-taking behaviors than native non-Hispanic whites. The mean number of risk behaviors was highest for Latino immigrants (1.78), followed by native-born Latinos (1.71), and native non-Hispanic whites (.99). A t-test revealed that non-Hispanic whites were statistically different from both Latino populations in either level of risk-taking behavior (p < 0.05). CONCLUSIONS: This sample of immigrant Latino students appeared to be vulnerable to engaging in risk-taking behaviors and did not exhibit the anticipated cultural protective factors associated with recent immigrants. The extent and variety of risk-taking behaviors clearly call for the development of culturally sensitive and accessible health education and health care services within a broader array of social, educational and support services. PMID- 7495833 TI - A soluble motor from the alga Nitella supports fast movement of actin filaments in vitro. AB - In the streaming cytoplasm of the Characean algae cell, the movement of organelles along actin bundles occurs at a striking rate of up to 60 microns s-1. To further characterize the molecular mechanisms responsible for this phenomenon, we have reconstituted the movement of actin filaments in vitro using defined biochemical components. We report that only a soluble cytoplasmic fraction devoid of organelles and filamentous material supports the movement of fluorescent labeled actin filaments on glass at a rate of up to 60 microns s-1. This fraction also contains the K(+)-EDTA ATPase and the actin-activated Mg2+ ATPase activities characteristic of myosin proteins. Therefore, on the basis of these observations, we conclude that Nitella cells have a soluble pool of non-filamentous myosin molecules with the mechanochemical properties expected for a motor responsible for cytoplasmic streaming in vivo. The preparation and conditions described here should be useful for the purification of this translocator. PMID- 7495834 TI - The site and stoichiometry of the N-phenylmaleimide reaction with myosin when weakly-binding crossbridges are formed in skinned rabbit psoas fibers. AB - Treatment of relaxed skinned rabbit psoas muscle fibers with 0.1 mM N phenylmaleimide (NPM) for 1 h locks all of the crossbridges in a weakly-binding state resembling that of the myosin.ATP crossbridge. Under these conditions, NPM reacts mainly with myosin heavy chain (Barnett et al. (1992) Biophys. J. 61, 358 367). Here the specific sites for that reaction are explored. Small bundles of rabbit psoas muscle fibers were treated with Triton X-100 to make the fiber sarcolemmas permeable. The bundles were treated with 0.1 mM [14C]NPM for 1 h, and homogenized for SDS-PAGE. 43 +/- 2.2% of the muscle fiber protein ran in the myosin heavy chain band, the same as for untreated fibers. An alkylating stoichiometry of 2.2 +/- 0.33 moles NPM per mole myosin heavy chain was determined. Exhaustive trypsin digestion followed by two-dimensional thin-layer chromatography and reverse-phase HPLC revealed two major sites on myosin heavy chain for NPM binding. The sites contained about the same amount of linked NPM, suggesting that the reaction stoichiometry of each site under the conditions studied is approx. 1 mol NPM/mol myosin heavy chain. Comparison of the labeled tryptic peptides with NPM-reacted synthetic SH1 and SH2 tryptic peptides and analysis of the treated fiber bundles' ATPase activity suggested that the sites for NPM reaction on myosin heavy chain when it locks crossbridges in a weakly binding state are Cys-697 (SH2) and Cys-707 (SH1). PMID- 7495835 TI - Changes in adult rat liver mitochondrial populations at different energy states analyzed by flow cytometry. AB - The present work studies the changes in green fluorescence intensity after Rh-123 staining of the low (LFP) and the high fluorescence populations (HFP) in isolated mitochondria from rat liver. The results show that the HFP represents a mitochondrial compartment less sensitive to changes in energy states. In addition, it is concluded that the use of Rh-123 to monitor changes in mitochondrial membrane potential should be undertaken with caution because, under certain circumstances, there is no correlation between the Rh-123 intensity of fluorescence due to its uptake by mitochondria and previously reported changes in the mitochondrial membrane potential. PMID- 7495836 TI - Characterization of an fdxN mutant of Rhodobacter capsulatus indicates that ferredoxin I serves as electron donor to nitrogenase. AB - A mutant of Rhodobacter capsulatus, carrying an insertion into the fdxN gene encoding ferredoxin I (FdI), has been studied by biochemical analysis and genetic complementation experiments. When compared to the wild-type strain, the fdxN mutant exhibited altered nitrogen fixing ability and 20-fold lower levels of nitrogenase activity as assayed in vivo. When assayed in vitro with an artificial reductant, nitrogenase activity was only 3- to 4-fold lower than in the wild type. These results suggested that the FdI-deleted mutant had impaired electron transport to nitrogenase. Immunochemical assay of both nitrogenase components showed that the fdxN mutant contained about 4-fold less enzyme than wild-type cells. Results of pulse-chase labeling experiments using [35S]methionine indicated that nitrogenase was significantly less stable in the FdI-deleted mutant. When a copy of fdxN was introduced in the mutant in trans, the resulting strain appeared to be fully complemented with respect to both diazotrophic growth and nitrogenase activity. Depending on whether fdxN expression was driven by a nif promoter or a fructose-inducible promoter, FdI was synthesized either at wild type level or in 10-fold lower amounts. The strain producing 10-fold less FdI did, however, display normal N2-fixing ability. Analysis of cytosolic proteins by bidimensional electrophoresis revealed that the fdxN mutant produced a 14 kDa polypeptide in amounts about 3-fold greater than wild-type cells. This protein was identified by N-terminal microsequencing as a recently purified [2Fe-2S] ferredoxin, called FdV, which cannot reduce nitrogenase. It is concluded that FdI serves as the main electron donor to nitrogenase in R. capsulatus and that an ancillary electron carrier, distinct of FdV, is responsible for the residual nitrogenase activity observed in the FdI-deleted mutant. PMID- 7495837 TI - Inhibition of Na+/K(+)-ATPase by phenoxyl radicals of etoposide (VP-16): role of sulfhydryls oxidation. AB - In the present work, we studied the effects of phenoxyl radicals, generated by tyrosinase-catalyzed oxidation of a phenolic antitumor drug, Etoposide (VP-16), on a purified dog kidney Na+/K(+)-ATPase by characterizing interactions of VP-16 phenoxyl radicals with the enzyme's SH-groups by ESR and correlating the loss of the enzymatic activity with the oxidation of its SH-groups, and oxidation of VP 16. VP-16/tyrosinase caused inhibition of Na+/K(+)-ATPase which was dependent on the incubation time and concentration of tyrosinase. The inhibition of Na+/K(+) ATPase was accompanied by a decrease of DTNB (5,5'-dithiobis-(2-nitrobenzoic acid)-titratable SH-groups. In the presence of Na+/K(+)-ATPase, a typical ESR signal of the VP-16 phenoxyl radical could be observed only following a lag period the duration of which was proportional to the concentration of the Na+/K(+)-ATPase added. Our HPLC measurements demonstrated that Na+/K(+)-ATPase protected VP-16 against tyrosinase-catalyzed oxidation. Combined these results suggest that redox-cycling of VP-16/VP-16 phenoxyl radical by SH-groups of Na+/K(+)-ATPase occurred. Ascorbate which is known to reduce the VP-16 phenoxyl radicals, protected the enzyme against inactivation, prevented oxidation of the enzyme's SH-groups. Reduction of VP-16 phenoxyl radicals by ascorbate was directly observed by the semidehydroascorbyl radical signal in the ESR spectra. VP-16 phenoxyl radical-induced oxidation of sulfhydryls and inhibition of the Na+/K(+)-ATPase may be responsible for at least some of its clinical side effects (e.g., cardiotoxicity) which can be prevented by ascorbate. PMID- 7495838 TI - Functional analysis of subunits III and IV of Bacillus subtilis aa3-600 quinol oxidase by in vitro mutagenesis and gene replacement. AB - Using the high efficiency of homologous gene recombination in Bacillus subtilis, a strategy for mutational analysis of the proton pumping aa3-600 quinol oxidase of this organism has been developed. The qox operon with the qoxA, qoxB, qoxC and qoxD genes, coding for the four subunits of this oxidase, was deleted and then replaced with mutated copies in which qoxC (subunit III) or qoxD (subunit IV) genes were deleted. The complete deletion of the qox operon caused disappearance of heme aa3-600 and a slight depression of the overall respiratory activity, compensated by alternative oxidase with no proton pumping activity. Deletion of qoxC probably resulted in a defective assembly of the aa3-600 quinol oxidase. The strain with deletion of qoxD gene expressed normal content of heme aa3-600 but exhibited a reduced respiratory activity and a significantly depressed proton pumping activity. These results show that subunit IV is critical for the activity of the proton pumping aa3-600 quinol oxidase. PMID- 7495839 TI - Developmental changes of the adenine nucleotide translocation in rat brain. AB - The perinatal development of the adenine nucleotide translocation in isolated rat brain mitochondria was studied. For that purpose the content of the adenine nucleotide translocase (ANT), the activity of adenine nucleotide translocation and the control of the ANT protein over State 3 respiration were estimated. From the newborn to the adult state there was a 4-fold increase in State 3 respiration which was paralleled by a 3-fold increase in the respiratory control ratio. The capacity of uncoupled respiration exceeded that of State 3 respiration in all developmental stages indicating that the activity of oxidative phosphorylation is influenced by that of ANT and/or ATP synthase. The content of the ANT protein, measured as bound pmoles of [3H]atractyloside per mg mitochondrial protein, increased more than 2-fold from birth to adultness in the first three postnatal weeks. The size of the exchangeable matrix (ATP + ADP)-pool was only slightly expanded during the same period. The translocation activity increased 2-fold from the newborn to the adult state and was a linear function of the ANT protein. Control of the ANT protein over State 3 respiration (quantified as flux control coefficient, CJoANT), was remarkable in brain mitochondria from newborn rats (CJoANT = 0.45 +/- 0.15), but declined during further development (CJoANT = 0.11 +/- 0.03, at the 20th day). The obtained results suggest that the postnatal enrichment of the ANT protein in rat brain mitochondria is an essential factor for the development of oxidative phosphorylation capacity in the early postnatal period. PMID- 7495841 TI - Physical (in) stability of liposomes upon chemical hydrolysis: the role of lysophospholipids and fatty acids. AB - As a consequence of chemical hydrolysis of liposomal phospholipids the organization of the lipid assembly can change from a lamellar into a micellar system. Different approaches provided evidence for this conversion: 31P-NMR analysis, turbidity measurements and ultracentrifugation experiments. Two conditions have to be met before this conversion can take place: (1) the liposomes must pass through a gel-to-liquid crystalline phase-transition during a heating or cooling run, and (2) the degree of chemical hydrolysis must exceed a critical hydrolysis percentage (or the phospholipid bilayer must contain critical amounts of lysophospholipid and fatty acid). As monitored by turbidity measurements, this critical level of hydrolysis and the relative change depended on the chain length and on the head group of the liposomal phospholipids. It does not depend on concentration, pH, storage temperature or on size of the liposomes within the experimental range. Addition of cholesterol to bilayers composed of dipalmitoylphosphatidylcholine prevents the lamellar to micellar transformation. Fluorescence anisotropy measurements of the lipophilic probe 1,6-diphenyl-1,3,5 hexatriene in 0.18-microns dipalmitoylphosphatidylcholine/dipalmitoylphosphatidylglycerol (10:1)-liposomes indicated that behavior of the probe below and above the phase-transition temperature was not affected by chemical hydrolysis, or even by formation of micelles. However, the phase-transition temperature range broadened and shifted towards higher temperatures upon hydrolysis. PMID- 7495840 TI - Proton/peptide cotransporter (PEPT 2) from human kidney: functional characterization and chromosomal localization. AB - We report here on the functional characterization of the H+/peptide cotransporter PEPT 2 cloned from human kidney and on the chromosomal localization of the PEPT 2 gene. PEPT 2, when functionally expressed in HeLa cells, induces the transport of the neutral dipeptide glycylsarcosine. The induced transport activity is markedly influenced by extracellular pH. The optimum pH for the transport process is 6.0 7.0. Kinetic analysis has revealed that PEPT 2 is a high-affinity transporter, the Michaelis-Menten constant for glycylsarcosine being 74 +/- 14 microM. The human intestinal H+/peptide cotransporter PEPT 1 has 4-fold less affinity for the dipeptide under identical experimental conditions. Studies with other chemically diverse dipeptides have established that PEPT 2 possesses higher affinity than PEPT 1 not only for neutral peptides but also for peptides consisting of anionic and/or cationic amino acids. Somatic cell hybrid analysis and in situ hybridization have shown that the gene encoding PEPT 2 maps to human chromosome 3q13.3-q21. PMID- 7495842 TI - The antitumor sulfonylurea N-(4-methylphenylsulfonyl)-N'-(4-chlorophenyl) urea (LY181984) inhibits NADH oxidase activity of HeLa plasma membranes. AB - Plasma membrane vesicles from HeLa S cells grown in culture bound with high affinity the antitumor sulfonylurea N-(4-methylphenylsulfonyl)-N'-(4 chlorophenyl)urea (LY181984). Based on binding site protection experiments with the radiolabeled thiol reagent N-[14C]ethylmaleimide, a ca. 34 kDa binding protein was identified. By analogy with a 36 kDa NADH oxidase from plant plasma membranes where activity was blocked by a growth-inhibitory herbicidal sulfonylurea, the sulfonylurea-binding protein of the HeLa plasma membranes has now been identified as a comparable sulfonylurea-inhibited NADH oxidase activity. The drug inhibited half maximally at about 50 nM which corresponded closely to the Kd for binding of [3H]LY181984 of 25 nM. A closely related but growth inactive sulfonylurea N-(4-methylphenylsulfonyl)-N'-(phenyl)urea (LY181985) inhibited the activity only weakly. The inhibition by LY181984 was analyzed kinetically and shown to be noncompetitive or uncompetitive depending on the concentration of NADH. With sealed right-side out plasma membrane vesicles, the NADH oxidase activity was about 90% inhibited by 1 microM LY181984. With frozen and thawed plasma membrane vesicles or with vesicles first solubilized with 1% Triton X-100, activity also was inhibited by LY181984 and not by LY181985 but the maximum inhibition at 10 microM LY181984 was about 50%. With plasma membranes from rat liver, neither LY181984 nor LY181985 affected the NADH oxidase even in the presence of detergent. Thus, selective inhibition or stimulation of the oxidation of NADH of tumor plasma membranes by the antitumor sulfonylurea LY181984 may be related to its antitumor activity. PMID- 7495843 TI - Paraquat2+/H+ exchange in isolated renal brush-border membrane vesicles. AB - The mechanism(s) by which paraquat (1,1'-dimethyl-4,4'-bipyridinium), a divalent organic cation (OC) and proximal tubule nephrotoxicant, crosses renal cell membranes is unclear. The structurally-related monovalent OC, 1-methyl-4 phenylpyridinium (MPP+), crosses the renal brush border via OC/H+ exchange using the same pathway by which tetraethylammonium (TEA) is transported. We examined whether paraquat shares the TEA(MPP+)/H+ exchanger by examining 14C-paraquat transport in rabbit renal BBMV. Compared to a pH equilibrium condition (pH 7.5in:7.5o), an H-gradient (pH 6in:7.5o) stimulated the 5 s and 60 s uptakes of 230 microM paraquat by 51% and 108%, respectively, and this stimulation was blocked by both 20 mM unlabeled paraquat and TEA. Pre-loading BBMV with 2 mM unlabeled TEA (under conditions of pH equilibrium) stimulated by 3-fold the 60 s uptake of 120 microM paraquat and by 5 min produced a transient intravesicular accumulation of paraquat that exceeded equilibrium (2 h) uptake by 45%. The presence of 200 microM paraquat in the extravesicular solution competitively inhibited H-gradient-stimulated transport of 14C-TEA in renal BBMV, increasing the apparent Kt for TEA transport from 169 microM to 379 microM, without significantly influencing the Jmax (16.0 vs. 15.4 nmol mg-1 min-1). The calculated Ki for paraquat (presumably equal to its Kt for transport) after transport was between 160 and 220 microM (depending upon the method of estimation). Significantly, the Kt for MPP+/H exchange is 12 microM, suggesting that the affinity of the exchanger is profoundly influenced by the presence on paraquat of a second positive charge. We conclude that renal transport of paraquat involves the OC/H+ exchanger of proximal cell luminal membranes and that this pathway may play a role in the renal secretion of polyvalent organic cations. PMID- 7495844 TI - Use of a novel method for determination of partition coefficients to compare the effect of local anesthetics on membrane structure. AB - A new, simple procedure for the determination of partition coefficients (P) was developed based on spectral effects caused upon addition of solutes to spin labeled model lipid membranes, and on the knowledge of their water solubility. Values of P were determined for nine local anesthetics (LA), amino-esters and amino-amides. The results were in good agreement with those found by phase separation and by a more complex, previously reported, methodology (Lissi et al. (1990) Biochim. Biophys. Acta 1021, 46-50) applied to either EPR or fluorescence spectra of probes incorporated in the bilayers. Both the present and the previously reported procedures make use of effects on membrane structure evaluated by spectroscopic techniques and offer the advantage of not requiring phase separation. The spectral effects, indicative of a decrease in bilayer organization increased with LA concentration, reaching a maximum at the drug water solubility, indicating that partitioning in the membrane is limited by saturation of the aqueous phase. A thermodynamic analysis of the partition data according to Hill (Hill, M.W. (1974) Biochim. Biophys. Acta 356, 117-124) showed that the LAs did not display ideal behavior. Knowledge of the partition coefficients allowed a comparison between effects at the same drug concentration in the membrane. Within a given family (esters, acyclic amides, cyclic amides) no clear proportionality was observed between effect and LA hydrophobicity, as reflected in the partition coefficient. Rather, the membrane perturbing ability is a result of steric effects originating in the mismatch between anesthetic and phospholipid shapes. PMID- 7495845 TI - Parameters influencing the introduction of plasmid DNA into cells by the use of synthetic amphiphiles as a carrier system. AB - Parameters that affect cellular transfection as accomplished by introducing DNA via carriers composed of cationic synthetic amphiphiles, have been investigated, with the aim to obtain insight into the mechanism of DNA translocation. Such insight may be exploited in optimizing carrier properties of synthetic amphiphiles for molecules other than nucleic acids. In the present work, the interaction of vesicles composed of the cationic amphiphile dioleyloxy-propyl trimethylammonium chloride (DOTMA) with cultured cells was examined. The results show that optimal transfection is dependent on the concentration of lipid, which determines the efficiency of vesicle interaction with the target cell membrane, as well as the toxicity of the amphiphiles towards the cell. A low lipid/DNA ratio prevents the complex from interacting with the cell surface, whereas at a relatively high amphiphile concentration the complex becomes toxic. Translocation efficiency is independent of the initial vesicle size but is affected by the size of the DNA. An incubation time of the DNA/amphiphile complex and cells of approx. 2-4 h is required for obtaining efficient transfection. In conjunction with observations on DNA/amphiphile complex-induced hemolysis of erythrocytes, a mechanism of DNA-entry is proposed which involves translocation of the nucleic acids through pores across the membranes rather than delivery via fusion or endocytosis. Dioleoylphosphatidylethanolamine, a phospholipid frequently used in a mixture with DOTMA ('lipofectin') strongly facilitates this pore formation. Translocation of the DNA is effectively prevented when the cells are pretreated with Ca2+ or pronase. These observations suggest that Ca(2+)-sensitive cell surface proteins play a role in amphiphile-mediated DNA translocation. PMID- 7495846 TI - Effect of current direction and K+ on polarization of the frog gastric mucosa proton pump. AB - When current was sent from serosa (S) to mucosa (M) across the frog stomach, there was a polarization (POL) of the open circuit potential (OCPD). POL was not affected by NaCl-free solutions, but was decreased by inhibitors of the H+ pump. In present experiments, current was sent to clamp the PD (VC) across the mucosa in steps of 20 mV up to 100 mV below the control OCPD, that is, current was sent from M to S. All experiments were performed in NaCl-free solutions. The POL was expressed as a % of the difference between the VC PD and the control OCPD. In 4 mM K+ control solutions, the POL was 11.8%; with 10(-3) M omeprazole (H+/K+ pump inhibitor), 1.1; with 10(-5) M SCH 28080 (H+/K+ pump inhibitor), 3.6; with 10(-3) M famotidine (H2 blocker), 1.6; and with 10(-2) M SCN-, 25.4 (inhibition of H+ sec, but not of the pump); in 79 mM K+ control solutions, 26.2; with 10(-3) M omeprazole, 4.2; with 10(-5) M SCH 28080, 15.9; with 10(-3) M famotidine, 5.6; and with 10(-2) M SCN-, 29.9. POL was higher in high K+ than in low K+ solutions contrary to what was observed in previous experiments with current sent from S to M. Results are explained on the basis of an electrogenic H+/K(+)-ATPase pump which includes a H+ channel, permeable to K+. With high K+ solutions, K+ is driven through the H+ channel onto the antiporter (ATPase) when current is sent from M to S, resulting in a greater POL of the pump. PMID- 7495847 TI - Ion selectivity of volume regulatory mechanisms present during a hypoosmotic challenge in vestibular dark cells. AB - Volume regulation during a hypoosmotic challenge (RVD) in vestibular dark cells from the gerbilline inner ear has previously been shown to depend on the presence of cytosolic K+ and Cl-, suggesting that it involves KCl efflux. The aim of the present study was to characterize hypoosmotically-induced KCl transport under conditions where a hypoosmotic challenge causes KCl influx via the pathways normally used for efflux. Net osmolyte movements were monitored as relative changes in cell volume measured as epithelial cell height (CH). A hypoosmotic challenge (298 to 154 mosM) in the presence of 3.6 or 25 mM K+ and loop-diuretics (piretanide or bumetanide) caused an increase in CH by about a factor of 1.2 presumably due to the net effect of primary swelling defined as osmotic dilution of the cytosol and RVD involving KCl efflux. A hypoosmotic challenge in the presence of 79 mM K+ and loop-diuretics, however, caused CH to increase by a factor of over 2.4. Presumably, this large increase in CH was due to the sum of primary and secondary swelling. Secondary swelling depended on the presence of extracellular K+ and Cl- suggesting that it involved KCl influx followed by water. The ion selectivity of secondary swelling was K+ = Rb+ > Cs+ >> Na+ = NMDG+ and Cl- = NO3- = SCN- >> gluconate-. Secondary swelling was not inhibited by Ba2+, tetraethylammonium, quinidine, lidocaine, amiloride, 4,4' diisothiocyanatostilbene-2,2'-disulfonic acid, 4-acetamido-4' diisothiocyanatostilbene-2,2'-disulfonic acid, 4,4'-dinitrostilbene-2,2' disulfonic acid, 5-nitro-2(3-phenylpropylamino)benzoic acid, acetazolamide, or ethoxyzolamide. These data define a profile of the hypoosmotically-induced KCl transport pathways. The ion selectivity and the blocker insensitivity are consistent with the involvement of the apical slowly activating K+ channel (IsK or minK channel) and the basolateral 360 pS Cl- channel. The involvement of these channels, however, remains to be demonstrated. PMID- 7495848 TI - Chlorsulfuron blocks 2,4-D-induced cell enlargement and NADH oxidase in excised sections of soybean hypocotyls. AB - Elongation of soybean hypocotyl sections induced by the auxin herbicide, 2,4 dichlorophenoxyacetic acid (2,4-D), was blocked by micromolar concentrations of the sulfonylurea herbicide chlorsulfuron (2-chloro-N-[[(4-methoxy-6-methyl-1,3,5 trizin-2- yl)amino]carbonyl]benzenesulfonamide). The inhibition at equimolar concentrations of 2,4-D and chlorsulfuron was overcome by a 10- or 100-fold excess of 2,4-D. Stimulation by 2,4-D of an 2,4-D-responsive NADH-oxidase activity of the soybean plasma membrane also was blocked by the presence of concentrations of chlorsulfuron equimolar to the 2,4-D. Chlorsulfuron alone was largely without effect on either cell elongation or the auxin-stimulated NADH oxidase activity over a range of chlorsulfuron concentrations. The results show a clear correlation between inhibition of auxin-stimulated cell elongation and the inhibition of an auxin-stimulated component of a plasma membrane NADH-oxidase activity. PMID- 7495849 TI - Quantification of the Na+/K(+)-pump in solubilized tissue by the ouabain binding method coupled with high-performance gel chromatography. AB - Membrane-bound Na+/K(+)-ATPase purified from dog kidney outer medulla was solubilized with octaethylene glycol n-dodecyl ether (C12E8) and incubated with [3H]ouabain in the presence of NaCl. ATP and MgCl2 for 10 min at 0 degrees C. The resulting enzyme was separated, by high-performance gel chromatography executed at 0.2 degrees C. Mainly into its (alpha beta)2-diprotomer and alpha beta protomer, which both bound stoichiometrically to [3H]ouabain. The amounts of ouabain that bound to the tissue itself and its microsomes could be estimated in the same way, as [3H]ouabain was found to bind only to the diprotomer and protomer they possessed. The amounts of ouabain that bound to them in the solubilized state were at least 5-times higher than those that did so when they were non-solubilized, suggesting that the surfactant rendered the enzyme accessible to ouabain. When the solubilized tissue (138 mg ml-1 wet tissue) was reacted with ouabain in the presence of 0.1 M NaCl and 4.8 mM MgCl2 for 10 min at 0 degrees C, maximal ouabain binding was attained in the presence of 18.3 microM [3H]ouabain, 1.2 mM ATP and 3 to 5 mg ml-1 C12E8, which was common to the outer medulla and human colon cancer cells. The present method enabled the pump number in protein and tissue samples in the range 7.2 x 10(-9) (purified pump) to 1.5 x 10(-12) (cancer tissue) mol/mg protein to be estimated within 2 h. PMID- 7495850 TI - pH modulates cAMP-induced increase in Na+ transport across frog skin epithelium. AB - Apical membrane potential (Va), fractional apical membrane resistance (FRa), and/or intracellular pH (pHi) were measured in principal cells of isolated frog (Rana pipiens) skin with microelectrodes under short-circuit conditions. Apical exposure to 0.33 mM 8-(4-chlorophenylthio)adenosine 3',5'-cyclic monophosphate (cAMP) depolarized Va, decreased FRa and increased short-circuit current (Isc). cAMP-induced 50% larger effects on Va and Isc at external pH (pHo) of 8.0 than at pHo 6.4. Increasing pHo from 6.4 to 8.0 in presence of cAMP further depolarized Va and increased Isc. cAMP-induced effects on Va and Isc were observed in the absence of Cl- and HCO3- and in the presence of 1 mM 4,4'-diisothiocyanostilbene 2,2'-disulfonic acid (DIDS) or 10 microM 5-(N-ethyl-N-isopropyl)amiloride (EIPA) or 1 microM 5-(N-methyl-N-isobutyl)amiloride (MIA). These data indicate that Na(+)-H+ exchange, Cl(-)-HCO3- exchange, and electrogenic Na(+)-(HCO3-)n cotransport are not involved in cAMP-induced increase in Isc. Apical exposure to 2 mM Cd2+ or Zn2+ depolarized Va, decreased FRa, increased Isc and increased pHi. In HCO(3-)-free solutions containing DIDS, unilateral replacement of apical Cl- by NO3- induced a fast transient depolarization of Va and an increase in Isc. These data suggest that potential-dependent changes in pHi are involved in increases in Isc. However, when changes in Va were minimized by pretreating the basolateral membrane with 25 or 75 mM K+, the cAMP-induced increase in Isc was not blocked. These data indicate that changes in pHi do not play a strict regulatory role but are only permissive in cAMP-induced effects on Isc. PMID- 7495851 TI - 2H-NMR study of two probe-labelled glycosphingolipid-derived signalling modulators in bilayer membranes. AB - We describe here the first report of sphingoid bases bearing non-perturbing 2H probe nuclei. These were produced, by two different routes of partial synthesis, to permit direct assessment of their arrangement and behaviour as minor components in membrane systems. Wideline 2H-NMR spectra of N,N dimethylsphingosine with deuterated amino-methyl groups ([2H6]dimethylsphingosine), and of lyso-dihydrogalactosylceramide (lyso-GalCer) with deuterium nuclei at C4,C5 of the sphingosine backbone and at C3,C4 of the galactose ring ([2H4]lyso-GalCer), were recorded in unsonicated, cholesterol containing fluid bilayer membranes. The sphingolipid metabolites behaved as single populations of lipid amphiphiles dispersed uniformly in the membrane and undergoing rapid symmetric motion about their long molecular axes. This was the case throughout the pH ranges examined, which included values generally considered for the cell cytoplasm. Spectra of [2H6]dimethyl sphingosine indicated that the methyl groups are equivalent on the NMR timescale, and that the molecule's orientation and behaviour are largely unaffected by pH over the range, 6 to 10.5. There was no spectral evidence of deprotonation of the tertiary amine function in this range. Similarly, variation of pH between 6.4 and 8.9 had virtually no effect on the average conformation and orientational order of lyso GalCer at the level of C4,C5 in the sphingosine backbone. pH did, however, exert significant control over the orientation of the galactose residue--the effect being most marked in the region of the sphingoid base pKa. The lyso-glycolipid showed some evidence of being less motionally ordered than the corresponding parent species, presumably as a result of removal of constraints imposed by the fatty acid. PMID- 7495852 TI - Stability against temperature and external agents of vesicles composed of archael bolaform lipids and egg PC. AB - The bolaform lipid PLE extracted from the thermophilic archaeon Sulfolobus solfataricus and its mixtures with egg phosphatidyl-choline (egg PC) have been used to prepare sonicated vesicles. The leakage of entrapped calcein was continuously monitored by fluorescence dequenching. The half times of leakage have been used to compare vesicle stability under different conditions of temperature, lipid composition and presence of destabilizing agents like Ca2+ ions and poly(ethylene glycol) (PEG). It has been found that leakage is primarily modulated by the monopolar/bipolar lipid ratio. In particular, the half time of leakage for vesicles formed from a mixture of the polar lipid extract (PLE) and egg PC is characterized by a maximum at about 1:2 molar ratio. The free energy of mixing has been evaluated from pressure-area isotherms on monolayers at the air/water interface. The results indicate a non monotonous behaviour of the excess free energy of mixing as a function of the molar ratio and the occurrence of a minimum at a fixed molar ratio. The possible formation of a complex is discussed and compared with previous calorimetric measurements on similar compounds. PMID- 7495853 TI - EPR study of spermine interaction with multilamellar phosphatidylcholine liposomes. AB - The interaction of spermine with egg-yolk phosphatidylcholine liposomes was investigated. The EPR spin labeling technique evidenced that spermine induces modifications of some membrane functions of biological interest like water permeability and is a possible modulator of diffusion processes for charged and polar molecules. The association constant for a hypothesized complex between spermine and the phosphate group of phosphatidylcholine was evaluated by enzymatic methods. PMID- 7495854 TI - Lysophosphatidylcholine inhibits vesicles fusion induced by the NH2-terminal extremity of SIV/HIV fusogenic proteins. AB - Intermediate lipid structures such as inverted micelles and interlamellar attachments are thought to play a crucial role in different biological processes like exocytosis, intracellular trafficking and viral infection. In the present study, we provide evidence that lipid mixing of large unilamellar lipid vesicles (LUV) mediated by the NH2-terminal sequence of the SIV gp32 and of HIV gp41 is inhibited by external addition of lysophosphatidylcholine (lysoPC) to LUV containing phosphatidylethanolamine in their lipid bilayer. Leakage experiments confirm that lysoPC enhances the stability of the lipids organization. The temperature dependence of the two processes as well as the complementary shape of PE and lysoPC suggest that the PE-lysoPC interaction is involved in the fusion inhibition and stabilization of the bilayer. PMID- 7495855 TI - Cloning and primary structure of a new hemolysin gene from Bacillus cereus. AB - A new hemolysin gene from Bacillus cereus VKM-B164 was cloned in Escherichia coli and sequenced. Deduced protein consists of 219 amino acids and has a molecular mass of 24.4 kDa. It has been concluded that the hemolysin, named 'hemolysin III', is distinct from the B. cereus hemolysins reported previously: cereolysin, sphingomyelinase, cereolysin AB, hemolysin II, and 'cereolysin-like' hemolysin (Honda, T., Shiba, A., Seo, S., Yamamoto, J., Matsuyama, J. and Miwatani, T. (1991) FEMS Microbiol. Lett. 79, 205-210). PMID- 7495856 TI - A secY homologous gene in the crenarchaeon Sulfolobus acidocaldarius. AB - The nucleotide sequence of an open reading frame, located upstream of the gene for adenylate kinase, was determined in the thermoacidophile crenarchaeon Sulfolobus acidocaldarius. Data bank searches identified the sequence as a secY homologous gene. The DNA derived protein sequence of total 463 amino acids contains 10 hydrophobic domains. A sequence alignment with other prokaryotic and eukaryotic secY sequences reveals significant homology, but the secY primary sequence of S. acidocaldarius shows only a low degree of similarity with the secY counterparts of the euryarchaea Methanococcus vannielii and Haloarcula marismortui. A transcription analysis indicates, that the secY gene is cotranscribed with the gene coding for adenylate kinase. PMID- 7495857 TI - cDNA cloning of mouse ferredoxin reductase from kidney. AB - A cDNA encoding ferredoxin reductase has been isolated from a mouse kidney cDNA library using human ferredoxin reductase cDNA as a probe. Mouse ferredoxin reductase coded for 494 amino acid residues. The mouse mature enzyme which comprises 460 amino acid residues shared 87.8-89.1% amino acid identities with the bovine and human enzyme. Northern blot analysis showed that ferredoxin reductase mRNA was expressed in the adrenal, testis and ovary and to a lesser extent in the liver and kidney. However, this mRNA in the adrenal cell line, Y-1 cell, was not induced by adenosine 3',5'-cyclic monophosphate (cAMP) in contrast with ferredoxin mRNA. PMID- 7495858 TI - Structure and sequence of the gene encoding the alpha-subunit of rat translation initiation factor-2B. AB - Eukaryotic initiation factor-2B (eIF-2B) is a heteropentameric guanine nucleotide exchange factor involved in the recycling of eIF-2 during the translation initiation phase of protein synthesis. The activity of eIF-2B is controlled by diverse mechanisms under a wide range of conditions. As part of our studies into the control of the expression of eIF-2B subunit mRNAs, we have isolated and characterized the rat gene encoding the alpha-subunit of eIF-2B. The gene is comprised of 9 exons which are contained within 8.5 kb of genomic DNA. A comparison of the 5'-flanking region of the gene with that of the previously isolated eIF-2B epsilon gene reveals that both possess a potential binding site for the alpha-Pal transcription factor as well as several other regions of homology. PMID- 7495859 TI - An aberrant lck mRNA in two human T-cell lines. AB - Low stringency screening of a human T-cell cDNA library with a c-src probe resulted in the isolation of several cDNAs of approx. 800 base pairs. Sequence analysis revealed that these clones encoded partial cDNAs for the src-family tyrosine kinase p56lck. A novel feature of this partial cDNA was that it contained a 93 bp intron insertion that was not observed in other full length or partial lck cDNA. This aberrant lck RNA was also detected in the mRNA pool by Northern blotting and PCR amplification of poly(A)+ RNA isolated from two human leukemic T-cell lines. PMID- 7495860 TI - An archaeal gene upstream of grpE different from eubacterial counterparts. AB - In some eubacteria with a dnaK locus in which grpE is close upstream of dnaK, grpE is preceded by an open reading frame (orf) believed to be a heat-shock gene. We also found an orf, orf16, upstream of grpE in the archaeon Methanosarcina mazei S-6, but this gene differs from the eubacterial counterpart: it is shorter, does not respond to a temperature upshift as heat-shock genes do, and the deduced protein Orf16, does not resemble the proteins coded by the eubacterial equivalents. orf16 is expressed monocistronically, with a transcription initiation site 24 bases upstream of the translation start codon, 22 bases downstream of a putative promoter identical to the consensus promoter for genes in methanogens. This initiation site is used by heat-shocked and non-heat-shocked cells in the two morphologic stages of M. mazei S-6 tested, i.e., packets and single cells. Three transcription termination sites were identified, one of which is detectable only in non-heat-shocked cells. Data from comparative analyses of the Orf16 deduced amino acid sequence and those of other known proteins, as well as the apparent biochemical characteristics of Orf16, suggest that the latter is a membrane molecule. PMID- 7495861 TI - Cloning and characterization of full-length cDNAs coding for the DNA topoisomerase II beta from Chinese hamster lung cells sensitive and resistant 9 OH-ellipticine. AB - DNA topoisomerase II beta cDNAs from Chinese hamster lung cells sensitive (DC-3F) and resistant to 9-OH-ellipticine (DC-3F/9O-HE) were isolated. In the sensitive cells, the sequence defines an open reading frame of 4839 nucleotides, and extends over a 323 nucleotides untranslated region up to the putative polyadenylation site. The deduced amino acid sequence predicts a protein with 1612 amino acids in length and a calculated molecular mass of approx. 182 kDa. The cDNAs from the resistant cells only differs by one mutation at position length and a calculated molecular mass of approx. 182 kDa. The cDNAs from the resistant cells only differs by one mutation at position 1710 which converts a Trp codon (TGG) to a stop codon (TGA). This mutation accounts for the loss of DNA topoisomerase II beta in the 9-OH-ellipticine resistant cells. PMID- 7495862 TI - Multiple nosZ promoters and anaerobic expression of nos genes necessary for Pseudomonas stutzeri nitrous oxide reductase and assembly of its copper centers. AB - Respiration of N oxides (denitrification) by bacteria is expressed facultatively in response to environmental stimuli. We have studied the transcriptional organization of the nos gene cluster of Pseudomonas stutzeri. This cluster carries the information for a functional nitrous oxide reductase (NosZ) which catalyzes the last step of the denitrification process. The nos genes are transcribed in three units, nosR, nosZ, and nosDFY. Transcription of nosZ is initiated from six different promoters which extend over a region of about 200 bp. The activity of two promoters varies subject to different growth conditions. Promoter P3 is active preferentially under denitrifying conditions and presumably under the control of a homolog of the transcriptional regulator FNR. Promoter P2 is the most active start site under aerobiosis and likely to initiate the low constitutive expression of nosZ. Transcription of nosR, encoding a regulator for nosZ expression, and transcription of the nosDFY operon, required for the copper chromophore assembly of NosZ, are both initiated from a single promoter. Transcription of nosR and the nosDFY operon was shown by phoA and lacZ fusions to be activated under a lowered oxygen tension and the simultaneous presence of an N oxide. The enzymatic activities associated with the hybrid proteins suggest for NosR and NosF a location in the cytoplasmic membrane and the cytoplasm, respectively. PMID- 7495863 TI - A 252 bp upstream region of the rat spermatocyte-specific hst70 gene is sufficient to promote expression of the hst70-CAT hybrid gene in testis and brain of transgenic mice. AB - The rat hst70 gene belongs to a heat shock hsp70 multigene family and its expression has been detected so far solely in spermatocytes. To investigate the cis-elements responsible for testis-specific expression of the hst70 gene we produced several lines of transgenic mice carrying fragments of the 5'-flanking regions of the hst70 gene fused to the chloramphenicol acetyltransferase (CAT) reporter gene. Hybrid genes of series B were constructed such that, besides the 780 bp, 343 bp and 163 bp 5'-flanking region these plasmids contained no other sequences of the hst70 gene. In hybrid genes of series D the CAT gene was ligated to 343 bp and 252 bp 5'-flanking regions together with the 57 bp of the 5'-end nontranslated (leader) sequences of the hst70 gene. We found that in 780/B, 343/B, 343/D and 252/D adult mice the transgene was specifically and highly expressed in testes. In developing testes the high CAT activity appeared in transgenic mice aged 3 weeks and older. None of the three 163/B transgenic lines exhibited CAT activity in any tissue analyzed. In all CAT expressing lines a weak but significant CAT activity (up to 5% of that in testis) was detected also in the brain. RNase protection assay confirmed that the endogenous hst70 gene transcripts are present in testis as well as in brain of nontransgenic rats and mice. Our data show that the cis-regulatory sequences responsible for testis specific and developmentally regulated expression of the hst70 gene are localized within the 252 bp region 5' to the gene and neither the 5'-end nor 3'-end nontranslated sequences of the gene are important for this specificity. PMID- 7495864 TI - Equilibrium model in an in vitro poly(ADP-ribose) turnover system. AB - Poly(ADP-ribose) metabolism plays an important role in numerous DNA-related functions. This homopolymer is synthesized by poly(ADP-ribose) polymerase and is degraded mainly by the poly(ADP-ribose) glycohydrolase. The activities of these two enzymes in the nucleus are closely coordinated. To better understand the interactions between these enzymes, we designed an in vitro system in which both enzymes are present at the same time. In this work, we report a model describing the synthesis and degradation of the poly(ADP-ribose) in turnover conditions. Because the half-life of the polymer in the cell is close to 1 min, we studied the very early kinetic interactions of these two enzymes. PMID- 7495865 TI - Eukaryotic topoisomerase II cleavage is independent of duplex DNA conformation. AB - Alternating purine-pyrimidine (RY) repeats have been identified in naturally occurring DNA and have many intriguing properties. Eukaryotic topoisomerase II displays significant cleavage activity at RY repeats (Spitzner et al. (1990) Nucleic Acids Res. 18, 1-11) due to the homology between RY repeat and the topoisomerase II consensus sequence. Cleavages are remarkably strong on duplex B form DNA. Certain RY elements are known to adopt altered DNA forms, such as Z DNA, under the influence of superhelical stress. To investigate the dependence of topoisomerase II activity on DNA conformation, a plasmid containing a 40 bp of deoxyguanine-thymine repeat was constructed and the dependence of topoisomerase II cleavage patterns were compared. Although the degree of negative supercoiling strongly affected the overall efficiency of topoisomerase II cleavage, the sequence specificity was identical over a wide range of superhelical densities. These results suggest that topoisomerase II site specific action on duplex DNA is largely independent of DNA conformation. Moreover, since the GT target sequence is known to adopt a Z-DNA structure under conditions of superhelical density used in these experiments, the results reveal that topoisomerase II is a DNA binding protein capable of recognizing Z-DNA structure in eukaryotic cell. PMID- 7495867 TI - Cloning and pharmacological characterization of a rabbit bradykinin B1 receptor. AB - A rabbit B1 bradykinin receptor cDNA was isolated from a rabbit aorta smooth muscle cell library. The 1223 bp cDNA clone encodes a protein of 352 amino acids which is 78% identical to the human bradykinin B1(3) receptor protein. Heterologous expression of the rabbit B1 receptor cDNA in COS-7 cells imparts a high affinity specific binding for 3H-labeled [des-Arg10,Leu9]kallidin. Scatchard analysis indicates that the receptor binds the radiolabeled ligand with a Kd of 0.5 nM. The ability of kallidin (Lys-bradykinin) and bradykinin analogues to compete with binding of 3H-labeled [des-Arg10,Leu9]kallidin was determined and defined a rank order of potency: [des-Arg10,Leu9]kallidin = [des-Arg10]kallidin > [des- Arg9]bradykinin = kallidin >> bradykinin. This receptor exhibits the classical B1 pharmacological property of preferentially binding to kinin analogues which lack the C-terminal arginine. In addition, the affinities for [des-Arg10]kallidin and [des-Arg10,Leu9]kallidin are 100-fold higher than those for the corresponding bradykinin analogues [des-Arg9]bradykinin and [des Arg9,Leu8]bradykinin which lack the N-terminal lysine. This pharmacological profile is characteristic of the B1 receptor subtype. PMID- 7495866 TI - Molecular cloning and sequence analysis of the rat liver carnitine octanoyltransferase cDNA, its natural gene and the gene promoter. AB - The full-length cDNA and the natural gene for rat peroxisomal carnitine octanoyltransferase (COT) have been isolated and sequenced. The 2681 bp long cDNA contains an open reading frame for 613 amino acids, resulting in a protein with a deduced molecular weight of 70,301, and a C-terminal peroxisomal targeting sequence (Ala-His-Leu). The isolated COT cDNA has 51 bp of the 5' untranslated region (UTR), 791 bp of 3' UTR, two putative polyadenylation sites, and a poly(A19-23) tail. Screening of a rat genomic DNA library in the lambda phage with the COT cDNA probe resulted in the isolation of seven overlapping clones, together containing the complete COT gene with seventeen exons. All of the exon intron boundary sequences conform to the GT-AG rule. The COT gene appears to spread over 40 to 60 kbp region of the rat genome. The transcription initiation site of the COT gene was determined through primer extension, and the promoter sequence up to the position -1140 was established. The promoter lacks the canonical TATA box and a promoter-reporter construct containing the sequence encompassing -1140 to +84 base positions and the firefly luciferase reporter cDNA yielded about 100-fold increase in promoter activity in transfected hepatoma cells. Some of the consensus sequences for putative cis elements present in the promoter sequence are: the two CCAAT motifs for CTF/NF1/CBP binding (at -284 and 93), two GC boxes for Sp1 binding (at -160 and -68), two AP2 sites (at -359 and 25), a half site (TGACCT) for the peroxisome proliferator activated receptor (PPAR) binding at -737 within a partial palindromic sequence region. Potential regulatory elements, such as several palindromes and repeat motifs for five different sequence segments, are also identified. PMID- 7495868 TI - Improved leishmanicidal effect of phosphorotioate antisense oligonucleotides by LDL-mediated delivery. AB - We have designed antisense oligonucleotides that can interact with lipoproteins in order to use them as vectors to facilitate the uptake by those cells expressing the corresponding receptor. Phosphorothioate (PS) oligonucleotides were linked at the 5' end to a palmityl group giving rise to PSPal conjugates. Such a modification enables the oligonucleotide to form a stable non-covalent complex with low density lipoproteins (LDL) through hydrophobic interactions. The antisense effect of LDL-oligonucleotide complexes was assayed by targeting the mini-exon sequence of Leishmania amazonensis in infected mouse peritoneal macrophages. A 16-mer antisense PSPal oligonucleotide/LDL complex exerted a more pronounced sequence-specific effect than the free oligomer: about 25% and 10% of infected macrophages were cured by a 48 h incubation in the presence of 2.5 microM of the complexed and the free oligomer, respectively. When oxidized LDL was used instead of the native one for complexation, a further 2-fold increase in the antisense effect was observed suggesting that alternative (unregulated) scavenger receptor can be used for more efficient delivery of antisense oligonucleotides into macrophages. In addition, a significant reduction of the parasitic load was observed in those cells that were not fully cured. PMID- 7495870 TI - Inactivation of the 5'-3' exonuclease of Thermus aquaticus DNA polymerase. AB - The gene for Thermus aquaticus (Taq) DNA polymerase enzyme (Taq Pol I) was mutagenized and sixty-two candidate clones were screened for enzyme activity. Two of the clones expressed enzymes (*Taq-3 and *Taq-5) that showed very reduced 5' 3' exonuclease activity and normal DNA polymerase activity. These two enzymes showed heat resitance and storage stability similar to Taq Pol I and had similar effectiveness in PCR. Processivity of the polymerases was compared by measuring the extension of an end-labeled primer annealed to a single stranded DNA, as well as by a PCR method. The processivity of *Taq-3 and *Taq-5 was similar to Taq Pol I (50-80 nucleotides) and more processive than a Taq Pol I deficient in the 5'-3' exonuclease due to absence of the first 290 amino acids (Stoffel fragment). The results indicate two amino acids which are required for normal 5'-3' exonuclease activity in Taq Pol I (Arg-25 and Arg-74). PMID- 7495869 TI - The choice between alternative IIIb and IIIc exons of the FGFR-3 gene is not strictly tissue-specific. AB - An essential feature of fibroblast growth factor receptors (FGFR) is the existence of multiple possibilities for alternative splicing. One of these concerns sequences of the mRNA coding for the C-terminal half of Ig domain 3 which corresponds to a part of the ligand-binding site. Two alternative exons, IIIb and IIIc, encode the C-terminal half of Ig domain 3. We show here that the alternative splicing choice between IIIb and IIIc exons of the FGFR-3 is not strictly tissue-specific: epithelial cells show exclusively IIIb transcripts while fibroblastic cells show a mixture of IIIb and IIIc transcripts. This is in contrast with the strictly exclusive alternative choice between IIIb or IIIc exons of the FGFR-2 gene: epithelial cells make only the IIIb choice while fibroblastic cells make only the IIIc choice. PMID- 7495872 TI - Mechanisms of iron uptake by mammalian cells. AB - On the basis of the discussion in this paper, the process of transferrin and iron (transferrin-bound iron and non transferrin-bound iron) uptake and transferrin release by reticulocytes are summarized diagrammatically. Although we were able to outline the pathways shown in the figure, there is still a long way to go before we achieve total understanding of the mechanisms of iron uptake. In addition, many important questions need to be answered. For example: what is the nature and properties of the iron carrier on the membrane? What is the relationship between the iron carrier and transferrin receptor? Is the iron carrier system in membranes of cells from different tissues similar or different? And how does iron cross the membrane of the endosomes after it is released from transferrin? All of these questions merit further investigation. PMID- 7495871 TI - The trypanosomatid Rieske iron-sulfur proteins have a cleaved presequence that may direct mitochondrial import. AB - We have cloned the gene that encodes subunit 4 of the T. brucei cytochrome-c reductase complex and a fragment of the C. fasciculata subunit 4 cDNA and have shown that subunit 4 is the Rieske iron-sulfur protein. The cleaved presequences of the trypanosomatid iron-sulfur proteins resemble conventional mitochondrial targeting presequences but are smaller than other eukaryotic iron-sulfur protein signal peptides. PMID- 7495873 TI - Distribution of A-kinase anchoring proteins in parietal cells. AB - Recent investigations have suggested that subcellular compartmentalization of second messenger responsive enzyme systems may be responsible for specific patterns of cellular activation. The type II cAMP-dependent kinase (A-kinase) is localized to particular subcellular domains through the binding of the regulatory subunit (RII) dimer to A-kinase anchoring proteins (AKAPs). Using a [32P]RII overlay assay, we have investigated the presence of AKAPs throughout the gastrointestinal tract, with specific emphasis focused on the gastric parietal cell. All gastrointestinal tissues contained at least one detectable AKAP (60 kDa), with five AKAPs (50-140 kDa) in fundic and antral mucosa. Isolated gastric glands contained four AKAPs. Two AKAPs (50 and 78 kDa) were detected in purified parietal cells, with the 78 kDa AKAP (AKAP78) specific to parietal cell enriched populations. RII-binding to all of these AKAPs was abolished by preincubation of [32P]RII with a synthetic peptide representing the RII-binding region of the AKAP, HT-31. AKAP78 was distributed throughout all membrane fractions of subfractionated parietal cells, with the largest amount of RII-binding detected in the light membrane fraction. Identification of A-kinase regulatory subunits by photoaffinity labeling with 8-azido-[32P]cAMP demonstrated that RII segregated into the same parietal cell subfractions as AKAP78. A majority (approximately 60%) of AKAP78 was detected in the Triton X-100-insoluble fraction, suggesting that this protein resides in a cytoskeletal domain. AKAP78 may be involved in localizing the type II A-kinase to specific intracellular locations in the parietal cell. PMID- 7495874 TI - Characterisation of cell movement by impedance measurement on fibroblasts grown on perforated Si-membranes. AB - Mouse fibroblasts grown on perforated Si-membranes (pore diameter approximately 10 microns have been studied to clarify cell locomotive ability. The cell motility was microscopically monitored by a time-lapse video system and, simultaneously, the impedance of the growing cells was measured every 5 s. The correlations between observed cell activities and measured impedance events are discussed and classified. The method is sensitive and allows discrimination between signals arising from translocation of single cells and those arising from filopodia activities. Both cell and filopodia motion could be detected. Designs of microdevices fabricated in semiconductor technology are presented. PMID- 7495875 TI - Copper uptake by cultured trophoblast cells isolated from human term placenta. AB - This paper has examined copper uptake from CuHis2 complexes by cytotrophoblast cells isolated from term human placenta. Uptake is time-dependent, reaching equilibrium after about 90 min, and saturable, with a calculated apparent Km of 0.174 +/- 0.061 microM and Vmax, measured over 30 min, of 0.721 +/- 0.092 pmol/min/micrograms DNA. To determine whether ATP was required for uptake, cells were incubated with inhibitors of glycolysis (iodoacetate) and the TCA cycle (sodium azide and cyanide). Iodoacetate and sodium azide had no effect on uptake, but cyanide decreased the initial rate of uptake. This effect was due to copper binding to the inhibitor and decreasing the effective substrate concentration rather than inhibition of uptake through ATP depletion. Ouabain and monensin had no effect, showing that neither the Na+ gradient nor endocytosis were involved in uptake. The monovalent ion chelator, bathocuproine sulphonate, had no effect on uptake but buthionine sulfoximine, an inhibitor of glutathione synthesis, did decrease both the rate of uptake and equilibrium copper levels, suggesting that copper may bind to glutathione within the cell. The data show that copper is taken up by a passive carrier-mediated transporter and, following uptake, binds to glutathione within the cell. PMID- 7495876 TI - Inhibition of prostacyclin release from cultured endothelial cells by nitrovasodilator drugs. AB - Pretreatment (18 h) of the bovine aortic endothelial cell line AG4762 to 500 microM sodium nitroprusside (SNP), glyceryl trinitrate (GTN) or 3-morpholino sydnonimine (SIN-1) significantly inhibited 100 nM bradykinin-stimulated prostacyclin (PGI2) release. SIN-1 produced the greatest reduction (67 +/- 6%), followed by SNP (47 +/- 12%) and GTN (45 +/- 9%). Only SIN-1 and GTN inhibited basal PGI2 release where again the effect of SIN-1 (66 +/- 6%) was greater than that of GTN (31 +/- 15%). There was no effect of SNP on basal PGI2 release. We have demonstrated this inhibition of bradykinin-stimulated PGI2 release is not the result of cell death. In addition, 8-bromo-cyclic GMP, whilst having no effect on basal PGI2 release, demonstrated a small but significant inhibition (15 +/- 6%) of the enhanced response to 100 nM bradykinin. These studies may reflect a mechanism by which the release of vasodilators from endothelial cells is altered during therapy with nitrovasodilators and thus may contribute to the development of tolerance to these drugs. PMID- 7495878 TI - Antibodies specific for proteolyzed forms of protein kinase C alpha. AB - The activation of protein kinase C (PKC) is irreversibly regulated by limited proteolysis catalyzed by a calcium-activated neutral cysteine protease, calpain. Calpain cleaves PKC alpha at specific sites in the hinge region between the catalytic and the regulatory domains of this kinase. Here we show a novel method for production of antibodies that bind specifically to the catalytic fragment of PKC alpha but not to the unproteolyzed protein. To detect proteolyzed PKC alpha 'cleavage site-directed antibodies,' which recognize amino-terminal regions in the nascent catalytic fragments and do not cross-react with the unproteolyzed enzymes, were raised using synthetic peptides corresponding to the amino-terminal sequences. The synthetic peptides used in this study were the sequences of human PKC alpha at the cleavage sites by m- and mu-types of calpains (LGPAGNKV and VISPSEDRKQPSNNLDRVKLT, respectively) and they are designated as CF alpha 2, CF alpha 4, in this order. Each synthetic peptide was injected into rabbit after conjugation with a carrier protein. The antibodies thus obtained (anti-CF alpha 2 or -CF alpha 4) specifically reacted with either the 46- or 45-kDa catalytic fragment of PKC alpha, respectively, whereas they did not cross-react with other fragments. Furthermore, the antibodies did not bind to the unproteolyzed enzyme nor fragments of PKC alpha obtained by treatment with other proteinases unless the fragment carried the same amino-terminal sequence. When human platelets were treated with calcium ionophore, the catalytic fragments of PKC alpha (45- and 46 kDa) were detected in the cytosol by immunoblotting with the antibodies. However, these antibodies did not bind unproteolyzed 80-kDa PKC alpha, although this form was dominant in the cytosol of the calcium ionophore-treated human platelets. In addition, the 45-kDa catalytic fragment of PKC alpha was detected in an apoptotic human fibroblast TIG-3 cells cultured in serum-free medium. Our method is applicable for analysis of proteolysis in various cellular states. PMID- 7495877 TI - Inhibition mechanism of HSP70 induction in murine FM3A cells maintained at low culture temperature. AB - We have shown previously that induction of HSP70 synthesis in murine FM3A and its mutant ts85 cells by heat shock is somehow modulated by culture temperature. In this study, we further examined activation of heat shock transcription factor (HSF) and induction kinetics of HSP70 synthesis and HSP70 mRNA in FM3A and ts85 cells maintained at 37 degrees C (37 degrees C-cells) and 33 degrees C (33 degrees C-cells). Upon exposure to heat shock, HSF was activated to a high level in 37 degrees C-FM3A cells, whereas HSF was activated only to a low level in the 33 degrees C-cells. The induction of HSP70 mRNA and HSP70 synthesis occurred successively in the 37 degrees C-cells but not in the 33 degrees C-cells. On the other hand, in both 37 and 33 degrees C-ts85 cells, activation of HSF, induction of HSP70 mRNA, and HSP70 synthesis occurred successively. Characteristically, protein synthesis in both 33 degrees C-FM3A and ts85 cells was significantly lower than in the respective 37 degrees C-cells, but constitutive HSP73 levels were similar among both the 37 and 33 degrees C-cells. Furthermore, inhibition of protein synthesis of FM3A cells did not influence the activation of HSF, but accelerated inactivation of the activated HSF. We discuss the possible inhibition mechanisms of activation of HSF in 33 degrees C-FM3A cells, regarding the function of HSP70 in both protein synthesis and repression of HSF. PMID- 7495879 TI - Activation of silent MDR1 genes in revertant cells by fusion with multidrug resistant cells. AB - We isolated revertant and resistant clones from multidrug-resistant K562/ADM cells and evaluated the expression of P-glycoprotein and the DNA copy number of MDR1. The 9 revertant clones contained 2- to 26-fold DNA copies of MDR1; however, they expressed an extensively decreased P-glycoprotein compared with K562/ADM, while the 10 multidrug-resistant clones contained 4- to 48-fold DNA copies, and the expression level of P-glycoprotein was dependent on the copy number of MDR1 DNA. The decreased expression of P-glycoprotein in the revertants was not due only to the loss of the copy number of MDR1 DNA. To elucidate the mechanism of P glycoprotein expression decrease in the revertants, a revertant clone (R1-5) was fused with a multidrug-resistant clone (A2-1) or with a drug-sensitive clone isolated from K562. Compared with K562 clone, the A2-1 contained 32-fold MDR1 DNA copies and showed 131-fold resistance to Adriamycin. The revertant clone R1-5 contained 26-fold MDR1 DNA copies but expressed only 5% the P-glycoprotein of A2 1 cells and showed only 2-fold resistance to Adriamycin. For selection of intraspecific hybrids, a neomycin-resistant or a blasticidin S-resistant gene was introduced into clones by electroporation of pSV2neo or pSV2bsr. The introduction of these resistant genes did not alter the copy number or expression of MDR1 in the clones. Hybrid cells between R1-5bsr and A2-1neo were found to express 136 +/ 15% of the P-glycoprotein of A2-1 cells evaluated by quantitive flow cytometry. These hybrid cells contained 41- to 48-fold MDR1 copies and showed the multidrug resistant phenotype, such as decrease of rhodamine123 accumulation and 120- to 210-fold resistance to Adriamycin (compared with K562), indicating that the 'silent' MDR1 genes in the revertant clone R1-5 were activated by cell fusion with an MDR clone. R1-5bsr x K562neo hybrids were found to contain 8- to 11-fold MDR1 copies and there was no increase in P-glycoprotein expression as compared with R1-5. PMID- 7495880 TI - A characterization of the chloride conductance in mesangial cells from the H-2Kb tsA58 transgenic mouse. AB - The chloride conductance of conditionally immortalized mesangial cells isolated from the H-2Kb-tsA58 transgenic mouse was studied in cells grown in permissive and non-permissive culture conditions. No differences were found in the magnitude of the chloride conductance in 140 mM tetramethyl ammonium chloride between cells grown in permissive and non-permissive culture conditions (1.08 +/- 0.05 nS and 1.02 +/- 0.05 nS). Selectivity sequences were defined as 1.8I- > 1.2Br- > lCl- and 1.3I- > 1.1Br- > lCl- respectively and both the anion channel inhibitors niflumic acid and ochratoxin A inhibited the chloride conductance in a dose dependent manner. The chloride conductance was made up from calcium-dependent and calcium-independent components and in the presence of 1 microM free calcium in the pipette this could be increased by the presence of 100 U ml-1 insulin added to the incubation medium. This appeared to increase the sensitivity of the conductance to levels of intracellular calcium as no differences were observed in the presence of either 1 mM or 100 nM pipette calcium. These data indicate that there is a significant chloride conductance in mesangial cells from the H-2Kb tsA58 transgenic mouse and minimal changes are observed with changes from permissive to non-permissive culture conditions. PMID- 7495882 TI - Voltage-activated and spontaneous Cl(-)-channel activity in cultured human melanoma cell-line A2058. AB - Voltage-activated and spontaneous chloride-channel activity was studied in melanoma cell-line A2058 by patch-clamp technique. Whole-cell and inside-out patch recordings carried out with leak subtraction show voltage-activated chloride-conductance. In addition, a large leak-type conductance typical of epithelial cells was found in whole-cell experiments. This current was carried mostly by chloride-ions but also a leak-type potassium conductance was found showing KCl fluxes to be possible. Cell-attached and inside-out patch recordings showed at least two types of spontaneous chloride-channel activity. Bursting, flickering-type channels were found only in cell-attached recordings. That led to the conclusion that some intracellular factors are needed for that kind of activity. A second spontaneous, 30 pS chloride-channel with slow kinetics was found both in cell-attached and inside-out patch configuration. A voltage activated chloride-channel found had a conductance of approximately 25 pS. In our experiments these channels did not need external calcium for activation. Voltage ramp recordings in cell-attached configuration gives the intracellular chloride concentration of 163 mM on the basis of chloride reversal potential (extracellular 146 mM in our experiments). Intracellular chloride concentration thus seems to be maintained slightly higher than the extracellular one. PMID- 7495881 TI - BAP2, a gene encoding a permease for branched-chain amino acids in Saccharomyces cerevisiae. AB - To select the gene coding for an isoleucine permease, an isoleucine dependent strain (ilv1 cha1) was transformed with a yeast genomic multicopy library, and colonies growing at a low isoleucine concentration were selected. Partial sequencing of the responsible plasmid insert revealed the presence of a previously sequenced 609 codon open reading frame of chromosome II with homology to known permeases. Deletion, extra dosage and C-terminal truncation of this gene were constructed in a strain lacking the general amino acid permease, and amino acid uptake was measured during growth in synthetic complete medium. The following observations prompted us to name the gene BAP2 (branched-chain amino acid permease). Deletion of BAP2 reduced uptake of leucine, isoleucine and valine by 25-50%, while the uptake of 8 other L-alpha-amino acids was unaltered or slightly increased. Introduction of BAP2 on a centromere-based vector, leading to a gene dosage of two or slightly more, caused a 50% increase in leucine uptake and a smaller increase for isoleucine and valine. However, when the 29 C-terminal codons of the plasmid-borne copy of BAP2 were substituted, the cells more than doubled the uptake of leucine, isoleucine and valine, while no or little increase in uptake was observed for the other 8 amino acids. PMID- 7495883 TI - Vascular smooth muscle cells metabolize endothelin-1 in the absence of a functional receptor. AB - Endothelin-1 (ET-1), a 21 amino acid vasoconstrictor peptide synthesized by vascular endothelial cells, exerts powerful actions on the underlying smooth muscle cells. The receptor and signal transduction mechanisms for ET-1 have been well characterized in rat aortic A10 vascular smooth muscle cells (A10VSMC). This investigation has characterized the internalization and metabolism of [125I]ET-1 by A10VSMC. A10VSMC internalized [125I]ET-1 rapidly in a receptor-mediated manner. However, inhibition of the binding/internalization had no effect on the metabolism of [125I]ET-1 by these cells. Thus, the presence of excess unlabeled ET-1 in the incubation, treatment of the cells with ET receptor antagonists, and homologous ligand-induced down-regulation of the ET-1 receptor all inhibited binding and internalization of [125I]ET-1 by A10VSMC, but not its metabolism. Furthermore, addition of excess unlabeled ET-1 to the incubations containing cells pretreated with the homologous ligand (receptor down-regulated cells) also failed to inhibit the metabolism of [125I]ET-1. Essentially similar characteristics of [125I]ET-1 binding and metabolism were exhibited by primary cultures of smooth muscle cells derived from rat thoracic aorta. Such ability of the vascular smooth muscle cells to degrade ET-1, which is produced constitutively by the endothelial cells, presents a novel mechanism in the regulation of its local and circulating concentration. PMID- 7495884 TI - Non-receptor cytosolic protein tyrosine kinases from various rat tissues. AB - Adipocytic-cytosolic non-receptor protein tyrosine kinase (CytPTK) when activated can substitute for the insulin receptor tyrosine kinase (InsRTK), in manifesting several insulin effects in insulin-receptor independent fashion. Our aims here were to utilize PolyGlu4Tyr, a good experimental exogenous substrate for protein tyrosine kinases (PTKs) in general, for studying qualitative and quantitative parameters of CytPTKs extracted from different tissue cytosols. At the same time, we would search for a unique specific marker specifically characterizing CytPTKs. High speed supernatants of spleen, thymus, smooth muscle, lung and kidney were found to be rich in CytPTK activities. Their specific activities being 6- to 13 fold that of liver or adipose cytosols. Brain, testis and adrenal cytosols were an intermediate source of CytPTK activity, whereas CytPTK activity of heart and skeletal muscle was low. It was also evaluated that the capacity of the cytosol to phosphorylate PolyGlu4Tyr is 15-50% that of the non-stimulated Triton X-100 extractable plasma membrane PTKs. Fractionation of the cytosols on superose 12 column revealed several CytPTKs within the same tissue, their peaks ranging between 30 and 450 kDa. Immunoblotting analysis showed Fyn and Lyn were present in most tissue cytosols. Upon immunoprecipitation, however, with anti-Fyn or anti Lyn, negligible amounts (< 2%) of the total cellular CytPTK were precipitated. Thus, these general markers of CytPTKs comprise only a minor proportion of the total intracellular PolyGlu4Tyr phosphorylating capacity. To see whether a specific marker for CytPTK could be detected, we also examined the requirement of CytPTKs for divalent ions, their preferred phosphate donor and their sensitivity to inhibition by known PTK inhibitors. We found that the order of reactivity with divalent cations was Co2+ > Mn2+ > Mg2+, while Zn2+ and Ca2+ did not support CytPTK activity. The best phosphate donor was ATP (ED50 = 5 microM), but other nucleoside 3-phosphates could substitute for ATP at high concentrations. With respect to these parameters, no basic difference exists between cytosolic and plasma-membrane PTKs. The PTK inhibitors, genestein and quercetin, inhibited both cytosolic and membranal PTKs at micromolar concentrations. In contrast, staurosporine was a potent inhibitor of CytPTKs (IC50 5-20 nM) and a poor inhibitor of membranal PTKs (IC50 10-40 microM). One of the conclusions we can draw from this study is that tissue cytosols contain PolyGlu4Tyr phosphorylating capacity in quantities greater than previously assumed and that the low level of phosphotyrosine found in cells is not the result of limited intracellular levels of CytPTKs. PMID- 7495885 TI - Cellular distribution of isoforms of protein kinase C (PKC) in pancreatic acini. AB - As in a previous study (Biochim, Biophys. Acta 1224 (1994) 127-138), we used quantitative immunoblot analysis and found that rat pancreatic acini possess four different isoforms of PKC-alpha, delta, epsilon and zeta. The phorbol ester 12-O tetradecanoyl phorbol 13-acetate (TPA) caused translocation of each isoform from the cytosol to the membrane fraction. CCK-8 increased diacylglycerol (DAG) and caused translocation of PKC-sigma and PKC-epsilon but not that of PKC-alpha or PKC-zeta. L-364,718, a CCK receptor antagonist, prevented as well as reversed the effects of CCK-8 on DAG and on translocation of PKC-sigma and PKC-epsilon. To explore the possibility that different isoforms of PKC might have different distributions in rat pancreas, we used immunocytochemistry to determine the cellular distribution of different isoforms of PKC in intact pancreas as well as pancreatic acini. In intact pancreas, PKC-alpha and PKC-sigma were detected in islet cells but not in duct or acinar cells. PKC-epsilon was detected in the apical region of acinar cells and PKC-zeta was detected over the luminal surfaces of acinar cells and the ductules that extend from the acinus. Neither PKC-epsilon nor PKC-zeta was detected in islets. In pancreatic acini PKC-alpha and PKC-sigma were detected in islets or fragments of islets that contaminated the preparation but were not detected in acinar cells. PKC-epsilon was detected in the apical region of acinar cells and adding 1 microM TPA or 1 microM CCK-8 accentuated the immunostaining but did not alter its cellular distribution. L-364,718 reversed the changes in immunostaining caused by CCK-8. PKC-zeta was detected over the luminal surface of the acinar cells. TPA, but not CCK-8 or CCK-8 followed by L 364,718, increased the number of acini that showed staining of the luminal surfaces of acinar cells. Thus, the present results demonstrate that different isoforms of PKC are distributed differently in rat pancreas and that the different patterns of distribution can explain, at least in part, the different responses to CCK-8. PMID- 7495886 TI - Some P2 purinergic agonists increase cytosolic calcium but not inositol 1,4,5 trisphosphate in isolated rat hepatocytes. AB - Based on the capacity to increase IP3 (inositol 1,4,5-trisphosphate), P2 purinergic agonists can be subdivided into two classes: ATP, ADP, UTP, 2deoxyATP, NAD and GTP significantly increased IP3 levels whereas ADP beta S, 2MeSATP, NADP, alpha, beta MeATP, beta, gamma MeATP and ATP alpha S had only a minor, non significant effect. Irrespective of their potency to increase IP3, all agonists were full glycogenolytic agonists and they all increased cytosolic calcium. With ATP and NAD, IP3 increasing agonists, and 2MeSATP and ADP beta S, non-IP3 increasing agonists, we found that the initial calcium response appeared to be an 'all or none' phenomenon, small amounts of the agonists being either ineffective or equally effective as high amounts. The minimal amount of an agonist needed to initiate a calcium increase and to promote glycogenolysis was very similar. In the absence of extracellular calcium, both groups of purinergic agonists (tested with ATP and 2MeSATP) were equally able to release calcium from intracellular stores. Cells with emptied intracellular calcium stores rapidly took up extracellular calcium upon treatment with ATP or 2MeSATP, the latter being the most potent. It seems therefore that all nucleotides tested increased cytosolic calcium and activated phosphorylase in a very similar way but some nucleotides had no effect on the levels of IP3. PMID- 7495887 TI - [Limits of rational public health policy]. PMID- 7495888 TI - [Coronary arteries and coronary artery bypass in CT: options, limitations and perspectives]. AB - Computed tomography examination of coronary arteries and aortocoronary bypass grafts would theoretically be a noninvasive alternative to the selective coronary angiography and could, therefore, potentially be used as a screening method for coronary heart diseases. Especially since the introduction of fast and ultrafast CT-methods (spiral-CT and electron beam-CT) many investigations have focused on this subject. On the basis of our own work as well as the experience of other authors, this article summarizes the options, limitations, and clinical relevance as well as the future perspectives of modern computed tomography techniques for the visualization of coronary arteries, coronary artery calcifications, myocardial perfusion, and aortocoronary bypass patency. PMID- 7495889 TI - [Radiation exposure in diagnostic CT: comparison between spiral CT and standard CT]. AB - CT-Dose-Index of Standard-CT corresponds to CT-Dose-Effect-Index of Spiral-CT. Spiral-CT provides a continuous data acquisition with well balanced dose profiles. Increasing pitch reduces radiation dose and nevertheless an unbroken reconstruction of scanned volume is possible. In contrast to Standard-CT, small parts at both ends of the volume scan are not reconstructed. Volume-Dose-Index therefore is elevated in short scan times but corresponds to that of Standard-CT in long scan times. 1 mm slices cause more radiation exposure due to their broadened dose profiles. PMID- 7495890 TI - [Use of multi-grid screens in intensive care units]. AB - Bedside chest radiographic examinations in intensive care units with grids and high kilovoltage-peak settings are impaired by artifacts caused by the angulation of the grid (grid cutoff). We have tried to optimize the use of grids in bedside chest radiographic examinations. The sensitivity to the effect of grid cutoff was examined in two grids with the same grid ratio (12:1) but different numbers of lamelle per cm (40 versus 70) using a water-phantom. The alignment of the grid in a bedside setting was optimised. 100 chest examinations in the intensive care unit were compared by five radiologists according to ten criteria. Grids with a high number of lamelle per cm (70) proved to be less sensitive to the effects of grid cutoff. By positioning the grid with the lamelle vertical to patient's body axis, asymmetric exposure of the lungs could be avoided. In addition, this setting allows an easy alignment of the grid by adjusting the grid together with the moveable part of the patient's bed. In examinations with grids, the advantages of high kilovoltage-peak setting were apparent. Bedside radiographic examinations in intensive care units with grids with a high number of lamelle and high-kilovoltage-peak setting can be performed without any major additional effort. By this method the image quality can be improved, especially in lungs with increased scatter-radiation. PMID- 7495891 TI - [Ultrasound detection of breast cancer with normal mammogram]. AB - In eleven patients with breast cancer the diagnosis was made by using ultrasound as the only method. In none of the patients could mammography show the malignoma. Although six of the patients underwent additional tube X-ray investigation, no radiological criteria of malignancy were found on the mammograms. In ten patients a lump of the breast was palpable and/or retraction of the skin was visible. In one woman, neither the clinical investigation nor the mammography could reveal the tumor. The ultrasound of the breast is a indispensable and useful method for diagnosing breast cancer. PMID- 7495893 TI - [CT and MRI in malignant peripheral neuroectodermal tumors]. AB - To evaluate the diagnostic accuracy of CT and MRI in the diagnosis of malignant peripheral neuroectodermal tumors (MPNT), the imaging data of nine patients with immunehistochemically proven MPNT were retrospectively analyzed. The average age was 24 years (ranging from 5 to 63 years). MRI and CT of all patients showed large tumors with direct infiltration of the surrounding structures. Intratumoral calcifications were not identified. All tumors showed equal to less high signal intensity in T1- and high signal intensity in T2-weighted images. Because of its high soft-tissue contrast, MRI is the most capable imaging modality in the diagnosis and therapy-monitoring in these tumors. PMID- 7495892 TI - [Results of catheter embolization of renal cell carcinoma]. AB - PURPOSE: We pointed out the value of catheter embolisation of kidney tumours. METHODS AND PATIENTS: We treated 42 patients with kidney tumours. For peripheral embolisation we placed Histoacryl or Ethibloc into the kidney artery via a transfemoral catheter. 33 patients had haematuria. 8 patients were lost for follow up. 26 patients were treated by embolisation alone and 8 patients were treated before nephrectomy. RESULTS: Haematuria was stopped in 98% and never reoccurred. The intraoperative blood-loss was 600 ml on average. 55% of patients treated by embolisation alone died within the first year. 51% of patients suffered from a slight postinfarction syndrome. One patient died of sepsis and one had pulmonary insufficiency due to AV shunt and pulmonary embolism. CONCLUSIONS: Peripheral embolisation of kidney tumours can be used successfully and is minimal invasive, in case of haematuria and pain. The perioperative risk is reduced due to limited blood loss. PMID- 7495894 TI - [Megadolichobasilar artery as a rare cause of hydrocephalus internus: synopsis of modern imaging methods]. AB - Megadolichobasilarartery (MDB), i.e. the widened, elongated and tortuous course of the basilar artery, has been the topic of numerous publications; about 350 cases have been reported world-wide. It can cause many symptoms; isolated or combined cranial nerve lesions and ischemic or hemorrhagic changes are the most frequent. A hydrocephalus internus is a rare occurrence and many patients do not exhibit any symptoms. To date, angiography, computed tomography, and to an increasing extent magnetic resonance imaging (MRI) are the principal methods for diagnosis of MDB. Angiographic-like representations with CT and MRI are further developments which represent an alternative to angiography. With the help of special MRI sequences, furthermore, non-invasive CSF flow measurements for the etiologic evaluation of a hydrocephalus can be performed. For the example of a patient with MDB and hydrocephalus internus, the possibilities of modern imaging techniques are presented and discussed. PMID- 7495895 TI - [Improved imaging of mandibular cysts using dental-CT]. AB - The purpose of this study was to evaluate orthopantomography and a newly developed CT software package (Dental-CT) in the preoperative assessment of mandibular cysts. In this study 16 cysts in 13 patients were examined radiographically by orthopantomography and dental-CT to reformat the axial scans into truly cross-sectional views perpendicular to the anatomical axis of the jaw. The obtained images were compared for visualisation of the alveolar canal, the mandibular anatomy and the cortical involvement. Dental-CT proved superior for establishing a correct radiographic diagnosis and was rated significantly (p < 0.001) higher with regard to examined points. Dental-CT has to be considered the method of choice in the preoperative evaluation of mandibular cysts providing detailed anatomical information needed for the further management. Radiation exposure remains within an acceptable range. PMID- 7495897 TI - [Pseudotumor calcinosis in renal failure]. AB - Tumoral Calcinosis is a rare clinical entity characterized by the presence of calcified, periarticular masses, as seen on radiographs. A severe case in a patient with chronic renal failure is presented and the complex etiology of this disease is discussed. PMID- 7495896 TI - [MR urography with the T2-weighted turbo-spin-echo sequence]. AB - Tumor-induced dilatation of the urinary tract is difficult to diagnose in the distal part of the ureter, including the stenosis, by ultrasound and X-ray. Often on account of renal insufficiency and allergy, i.v.-contrast media cannot be used. The present study should show the suitability of fast T2-weighted (turbo-) spin-echo sequences (T2-TSE) for MR-urography (MRU). Seven patients (62.3 +/- 6.1 years) were examined in the coronal plane with T2-TSE sequence (TR = 4500 ms, TE = 160 ms) and an MRU was calculated by using the MIP method (maximal intensity projection). This technique enabled urogram-like morphological representation of dilated urinary tract including stenosis in 6 of 7 patients. Assuming a high magnetic field homogeneity, MRU by using a T2-TSE-sequence, without i.v.-contrast media administration, can visualize the urinary tract dilatation and localize tumor-induced stenosis. PMID- 7495898 TI - [Gangliocytic paraganglioma of the duodenum]. AB - A duodenal gangliocytic paraganglioma was found to be the cause of nonspecific abdominal complaints in a 68-year-old man with a known carcinoma of the rectum and the prostate. The duodenal gangliocytic paraganglioma is generally regarded as being benign. In this case it is shown to be an additional tumor in a patient with two pre-existent malignancies. PMID- 7495899 TI - [Isolated dislocation of the os triquetrum--a rare wrist injury]. AB - The isolated traumatic dislocation of the triquetrum is an extremely rare injury. The clinical signs are unspecific. Roentgenograms of the wrist in posteroanterior and lateral views, possibly complemented by oblique views, will help to provide the diagnosis. The best therapy appears to be the open reduction and fixation with Kirschner wires. With the presented case report, the difficulties in using the classification carpal injuries are discussed. Finally, corresponding to the carpus ring theory, the plausibility of additional injuries to carpal ligaments in such trauma is pointed out. PMID- 7495900 TI - [Effects of the medical products legislation on roentgen services]. AB - On January 1, 1995 the law concerning medical products (Medizinproduktgesetz-MPG) came into force. It essentially regulates the production, circulation, and the putting into service of medical products as well as equipment. Among other things, the so-called active medical products include medical X-ray equipment whose aspects concerning atomic law remain covered by the radiation protection regulation for X-ray sources (RoV). Parallel to the coming into force of the MPG, the RoV was modified (Section 50, MPG), which in future might result in considerable consequences for the manufacturer, the operator, and the authorities. PMID- 7495901 TI - [Effect of solar activity on the physiological rhythms of biological systems]. AB - The infradian components, including those with a period of about 3.5, 7, 30 days, are ubiquitously found in biology, from unicells to complex biological organisms. It can be hypothesized that heliogeophysical factors other than the solar visible light, likely at the origin of the circadian system (with a period of about 24 hours), may be responsible for the infradian biosystems. PMID- 7495902 TI - [Dynamics of the morphofunctional status of cell cultures exposed to variations in the geomagnetic field at high latitudes]. AB - The goal of this research is to study a dependence between dynamics of the biological parameters, characterized the state of the cell cultures, and variations of geomagnetic field (GMF) in the quiet and disturbed conditions. The investigations were carried out on geophysical recording site in high-latitude regions. Geophysical and biological observations were conducted simultaneously and at the same place. Continuous cell lines of mouse, hamster and pisces were used in experiments. Morpho-functional characteristics, including the percentage of polynuclear cells were estimated. Has been shown that dynamic of morphofunctional state of cell culture associated with GMF variations. Geomagnetic activity (GMA) abrupt change is followed by a stepwise changes the state of cell cultures. Changes is properties of cell surface are related to cell fusion, the appearance of heterokaryons, enlargement of cellular adhesion and aggregation. Enhancement of morpho-functional variety in cellular populations involving a reorganization of nuclear material demonstrate a possible regulatory and evolutionary part of GMF variations. PMID- 7495904 TI - [The effect of geomagnetic disturbances in capillary blood flow in ischemic heart disease patients]. AB - A total of 80 patients with ischemic heart diseases (men-47, women-33) were followed up daily during 2-3 weeks. We studied capillary flow in the cuticle above the nail (eponichium) with TV-capillaroscope, allowed to conduct prolonged studies. We evaluated capillary indices for perivascular edema, erythrocytes aggregation, blood velocity. Microcirculations data were compared daily values of geomagnetic activity (A-index), three-hour-range indices (K) and atmospheric pressure. In the first day magnetic storms pathological changes of capillary flow were detected in 71.5% patients with acute myocardial infarction (men-73.7%, women-69.2%). We could see appearance of perivascular edema, red blood cell aggregation, delay and slowing down capillary flow. Similar changes were detected in patient with angina pectoris in 64.8% (men-73.3%, women-56.3%). Number patients with ischemic heart diseases reacted upon geomagnetic disturbances exceed more then 2.5 times quantity patients, who react upon change of atmospheric pressure. PMID- 7495903 TI - [Daily observations (1970-1992) of fluctuations in frequency of occurrence of a sector structure in bacterial colonies selected from open air and from S. aureus cultures]. AB - The frequencies of sector structure occurrence in different bacteria colonies (SSC) denuded from open air every day in period from 1970 to 1982 years and also in laboratory cultures Staphylococcus Aureus from 1984 to 1992 were investigated. The value SSC was expressed in percents to general number of colonies. Variations of average SSC is found out distinctly expressed opposition to 11-years cycle of Solar activity for the same period of time. The year cycle SSC was registered also with local maxima in June, August and November and global minimum in March. SSC also were observed on level of sea, on height about 1000 m over the level of sea and in gallery under the rock of thickness 1400 m (Gran Cacco, National laboratory, Assergi). The value of SSC has its maximum on height 1000 M and its minimum in gallery under the rock; it is possible, SSC depends on the intensity of space galactic rays. The results of experience, executed in April-July 1992 on cultures S.aureus have compared with average (on ten-day time periods) values for the same months of 1984-1991. Good correlation of these two curves of SSC was received. Array of measurements received from 1970 to 1992 and made conclusions can be used for long-term forecasts of cosmophysical fluctuating phenomena at least on latitude between Rome and Milano. PMID- 7495905 TI - [Effect of geophysical factors on indicators of cardiac sinus rhythm structure in children]. AB - Influence of geomagnetic activity, atmospheric pressure, temperature and humidity on heart rhythms parameters and wave structure was examined by statistic methods. Correlative relationships with different trends took place between some geophysical factors and child's heart rhythms indexes. PMID- 7495906 TI - [The role of gravitational tidal changes in regulating the systemic response of circulation and respiration to orthostasis]. AB - The transient reply parameters of passive postural changes (lying-sitting) were evaluated. The most important 8 variables of circulation and respiration were analysed during 20-30 min of orthostatic test. The findings were based on 552 tests performed on 237 volunteers. It was shown that the adaptation of a human organism to the postural hydrostatic factor is formed by the systemic cooperation of circulation and respiration like a hydraulic homeostat. Influence of some physical factors (physiotherapy) changed this cooperation. The same data is discovered in orthostatic responses as a result of the different phases of lunar semidiurnal gravity waves. That confirmed the gravity tidal variations have influence on human regulation systems. PMID- 7495907 TI - [Common periodicity of solar atmosphere rotation and function of the human body]. AB - The study is made of the occurrence rates of menstrual cycle periods for approximately 2000 women. Peaks on the distribution histogram, corresponding to 21, 25, 28 and 30 days, coincide with a set of axial rotation periods of the solar atmosphere. It is proposed that the functioning of human organism is determined not only by the Moon but by the rhythmics of solar system. PMID- 7495908 TI - [Changes in medical indicators during heliogeophysical disturbances]. AB - Influence of heliogeophysical factors on medical indexes is studied. Relationship between changes of these indexes and solar flares, magnetic and ionospheric storms is analyzed. Possibility of forecasting the biotropic effects by using the starting of heliogeophysical events as predictions is showed. Comparisons between calculations and experimental data are provided. Reasonable agreement was obtained. PMID- 7495909 TI - [Statistical approach to the validity of a diurnal cycle with periods of 4-6 hours]. AB - Confidence coefficient and reliability of previously unknown phenomenon of the interdian cyclicity (with the periods about 4-6 hs) in dynamics of physiological and pathological processes is shown. Discovery of critical time instances (at 2, 9, 14, 18, 22 o'clock of local "winter" decree time) of human life poses a problem of an application of this phenomenon to the solution of different medical and social problems. PMID- 7495910 TI - [Effect of gravitational tidal changes on the periodicity of T-lymphocyte receptor expression in vitro]. AB - The hourly registration of the expression of the human s T-lymphocytes subpopulations receptors in vitro during 6 hours demonstrated the existence of the periodicity from 2 to 4 hours. There is high degree of T-lymphocytes expressions coincidence from different individuals depending of the date. For most of daily average individual chronograms, there were described back linear and valid correlations with a character of heterogeneity of the gravity force changes. The results are in according to suggestion about the time field geometric effect to structural-functional cells periodicity. PMID- 7495913 TI - [Effect of solar and geomagnetic activity on population dynamics among residents of Russia]. AB - Basing on the annual data of mortality and birth-rate in Russia in 1867-1897 an attempt to evaluate the role of solar (sunspot number W) and geomagnetic activity (Kp-index) in the population was made. It was shown that mortality is affected by the solar activity and by the absolute value of geomagnetic activity deviation from the mean level: Mi = 3.29 + 0.0044Wi + 1 + 0.030 delta Kp,i (r = 0.78). For the birth-rate the following expression was obtained: Bi = 5.00-0.049 delta Kp,i + 1-0.0012Wi + 3; (r = -0.67). The correlation coefficients between the observed variations of the mortality (and birth-rate) and indices Mi and Bi amounts the values 0.78 and -0.67 correspondingly. PMID- 7495912 TI - [The connection between multiday rhythms of certain indicators of systemic and cerebral hemodynamics and autonomic function with cosmophysical rhythms in patients with essential hypertension]. AB - Near-week and multiple of them rhythms are revealed in the dynamics of the studied biological functions and cosmophysical indices. The conception of endogenic nature of biological rhythms is discussed. It is shown that the group of people sick with essential hypertension is divided into three subgroups according to the type of link of biological rhythms with the cosmophysical; hypersynchronous, moderate synchronous, and asynchronous. It is suggested that the type of link with external factors is attributed by the of interaction of various function in the organism. PMID- 7495911 TI - [Effect of electromagnetic fields of natural and anthropogenic origin on the frequency of pathology occurrence in St. Petersburg]. AB - The effect of man-made electromagnetic fluctuations and strong geomagnetic disturbances on human pathology-data from St. Petersburg (Russia, 1981) is analyzed. The most remarkable effect is the 7-day variation of the ambulance-call data for myocardial infarction, that show a decrease in pathology-rate (70%) during week-ends and public holidays. Results of measurement of man-made electromagnetic fluctuations in the frequency range 0.005-10 Hz in 1991 and 1994 show a big decrease in electromagnetic noise during week-ends. We suggest that very big 7-day variation in infarction rates is connected with the decrease of electromagnetic noise during Saturdays-Sundays. This phenomenon is responsible for the very big 7-day variation in infarction rate. The myocardial infarction rate cleaned up by meteorological and social effects show the increasing by a factor 1.14 during geomagnetic storms. PMID- 7495914 TI - [Temporal organization of living organisms and the problem of reproducing results of magnetobiological studies]. AB - The analysis of bad reproduction of experimental magnetobiological results was realized onto experimental data for extremely low frequency magnetic field influence on leucocytes level in animal blood. It was shown that one of the causes of bad experimental results reproduction is a dynamism of temporal organization parameters in animals that used as a biological control. PMID- 7495915 TI - The treatment of persistent night terrors with autogenic training: a case study. AB - This is a case study of a 16-year-old male with a long history of persistent and severe night terrors. After a comprehensive medical and psychological evaluation, the patient was trained in standard autogenic training exercises and an organ specific formula. By the end of the eight-week treatment, the nocturnal episodes ceased. Information at 11 weeks and 37 weeks follow-up supported the effectiveness of the treatment. PMID- 7495917 TI - A provisional model to predict blood pressure response to biofeedback-assisted relaxation. AB - Blood pressure (BP) response to biofeedback-assisted relaxation is not uniform among hypertensive individuals. The purpose of this exploratory study was to determine if selected psychophysiological variables could be used to identify individuals able to lower blood pressure using biofeedback-assisted relaxation. Responders were defined using a preset criterion of 5 mm Hg or greater decrease in mean arterial pressure. A logistic regression model derived from five variables (heart rate, finger temperature, forehead muscle tension, plasma renin response to furosemide, and mean arterial pressure response to furosemide) provided significant predictive power for BP response, exhibiting a sensitivity of 84.6% and a specificity of 80.0%. With future validation, the proposed model may provide useful information to identify patients likely to benefit from biofeedback-assisted relaxation. PMID- 7495916 TI - Neurotherapy for stroke rehabilitation: a single case study. AB - A 55-year-old male subject was treated with a two-tiered neurotherapy approach for a period of six months beginning approximately one year after a left-side CVA. Medical evaluation revealed left posterior temporal/parietal infarctions secondary to occlusion of the left internal carotid artery. The patient complained of hesitant speech with word finding difficulty and paraphasia, difficulty focusing his right eye, lack of balance and coordination, poor short term memory, poor concentration, anxiety, depression, and tinnitus. A quantitative electroencephalograph (QEEG) analysis revealed increased left-side 4 7-Hz activity and alpha persistence on eye opening. Two neurotherapy approaches were used beginning with electroencephalographic entrainment feedback (EEF). This was followed by neurofeedback to inhibit 4-7 Hz and increase 15-21 Hz over sensorimotor and speech areas. At the conclusion of treatment there were significant reductions in slow-wave activity. Improvement was evident in speech fluency, word finding, balance and coordination, attention, and concentration. Depression, anxiety, and tinnitus were greatly reduced. PMID- 7495918 TI - Operant (biofeedback) control of left-right frontal alpha power differences: potential neurotherapy for affective disorders. AB - Two experiments were done with subjects from a paid pool of undergraduates. In each study, there were five 1-hour sessions on each of 5 days: (1) Baseline: Rewards given for randomly selected 20% of the 700-ms sequential epochs; mean and SD of baseline power differences determined. 2) Exploration: Subjects were rewarded when right minus left alpha differences in an epoch were greater than the baseline mean plus about .85 SD (p = .20); subjects told to discover how to generate rewards. (3)-(5). Training: Subjects were paid (over and above the $8/h flat rate) in proportion to their hit rates. In the first study (in which active filters passed 8-12 Hz activity, and the rectified, integrated amplitude was utilized), 6 of 8 subjects met learning criteria (a significant difference between baseline and training scores). In the second study (in which on-line FFTs were used to extract alpha power), 3 of 5 subjects met learning criteria. PMID- 7495919 TI - Decreases in health care resource utilization in patients with rheumatoid arthritis following a cognitive behavioral intervention. AB - We previously reported the efficacy of a multicomponent cognitive-behavioral intervention including biofeedback to decrease pain, affective distress, and objective measures of disease activity in patients with rheumatoid arthritis (RA). In the present article we report evidence that this intervention is associated with reductions in RA-related clinic visits and days hospitalized as well as reductions in the costs of these medical services. Data were independently and objectively collected over an 18-month interval as part of a controlled group outcome study. The importance of documenting economic as well as clinical benefits of our treatments in specific patient populations is noted. PMID- 7495920 TI - Drug development and the deficit syndrome of schizophrenia. PMID- 7495922 TI - Language comprehension in schizophrenia: trait or state indicator? AB - Language comprehension, measured by the Luria-Nebraska Relational Concepts Factor Scale, was evaluated twice in 15 male DSM-III-R schizophrenic patients during a controlled double-blind haloperidol maintenance (without anticholinergics) and placebo replacement protocol. Fifteen male normal controls were tested once. Patients and controls were matched on age and education. Language comprehension was significantly reduced in patients under both pharmacologic conditions, as compared with controls. Patients' comprehension accuracy did not differ significantly between neuroleptic-treatment and placebo replacement conditions. Patients' comprehension accuracy was independent of positive symptoms, anxiety depression, measures of clinical course, and CSF and plasma monoamines. Comprehension accuracy was also not associated with patients' educational level or WAIS-R measures of their intellectual and short-term memory functioning. Patients' comprehension performance was significantly associated only with the negative symptom anhedonia-asociality during haloperidol maintenance. Thus, language comprehension in schizophrenic patients was independent of changes in pharmacologic treatment and the positive symptoms of psychosis. Results suggest language comprehension may represent a stable or trait characteristic in schizophrenia. PMID- 7495921 TI - Asymmetry of temporal lobe phosphorous metabolism in schizophrenia: a 31phosphorous magnetic resonance spectroscopic imaging study. AB - In vivo 31Phosphorous magnetic resonance spectroscopic imaging (31P MRSI) was performed on 18 chronic schizophrenic patients and 14 normal controls to determine if there was asymmetry of high-energy phosphorous metabolism in the temporal lobes of schizophrenic patients. Temporal lobe phosphorous metabolites were also correlated with severity of psychiatric symptomatology as assessed by the Brief Psychiatric Rating Scale (BPRS). Schizophrenics demonstrated significantly higher right relative to left temporal phosphocreatine/adenosine triphosphate (PCr/ATP), phosphocreatine/inorganic phosphate (PCr/Pi), and PCr as well as significantly lower right relative to left temporal ATP. There were no asymmetries of temporal lobe phosphorous metabolites in the control group. In addition, both left temporal PCr and the degree of asymmetry of temporal lobe PCr were highly correlated with the thinking disturbance subscale of the BPRS. This study provides further support for temporal lobe metabolic asymmetry in schizophrenia and its possible association with clinical symptoms. PMID- 7495923 TI - Cytokine production in drug-free and neuroleptic-treated schizophrenic patients. AB - A line of evidence indicates changes of the immune system in schizophrenic patients. We investigated the production of cytokines by peripheral blood mononuclear cells (PBMCs) in drug-free and neuroleptic-treated schizophrenic patients compared to healthy, normal controls. A significant reduction in interleukin (IL)-2 production was detected in untreated schizophrenic patients ( 59.6%; p < .05) as well as in IL-3-like activity (IL-3-LA) production (-27.4%; p < .05) in treated patients compared to controls. No alteration was observed in IL 1 beta production. It seems that schizophrenia is associated with diminished IL-2 production, while neuroleptic treatment interferes with the capacity of immunocompetent cells to synthesize and/or release Il-3-LA. The alteration in cytokine production did not correlate with either the severity of the disorder or the serum prolactin levels. PMID- 7495924 TI - Quantitative EEG in never-treated schizophrenic patients. AB - To clarify whether patients with schizophrenia still show EEG slowing in the absence of psychopharmacological treatment, EEG was analyzed in 20 acute never treated schizophrenics and 20 age-matched healthy controls using the computerized wave-form recognition method. Compared to controls, schizophrenics had more fast theta (6-8 Hz) and slow alpha (8-9 Hz) activity, and less fast alpha activity (9 13 Hz). The average EEG frequency at O1 correlated negatively with total and positive symptom scores on the BPRS in the schizophrenic group. These findings confirm that the frequency of alpha rhythm is slowed in schizophrenia and that this slowing is possibly related to the expression of psychopathology in this disorder. PMID- 7495925 TI - Effect of pindolol pretreatment on MK-212-induced plasma cortisol and prolactin responses in normal men. AB - Previous reports based on studies with serotonin (5-HT) precursors or direct acting agonists have suggested that postsynaptic 5-HT1A and 5-HT2A/5-HT2C receptors may stimulate cortisol and prolactin (PRL) secretion in man. To further clarify the role of these receptors in the regulation of cortisol and PRL secretion in man, the effects of 6-chloro-2-(1-piperazinyl) pirazine (MK-212), a centrally acting direct 5-HT2A/5-HT2C agonist, on the above hormones were studied in 11 normal men with and without pretreatment with pindolol, a 5-HT1A partial agonist. MK-212 induced a significant increase in plasma concentrations of cortisol and PRL. The MK-212-induced response in plasma cortisol was not diminished by pindolol pretreatment, whereas the MK-212-induced PRL response was significantly inhibited by pindolol pretreatment. These data suggest that the MK 212-induced cortisol response may be mediated by 5-HT2A or 5-HT2C receptor activation, or both, despite 5-HT1A inhibition; however, PRL secretion by MK-212 requires 5-HT1A receptor availability as well as 5-HT2A/5-HT2C receptor activation, since blockade of the former appears to blunt the PRL responses to MK 212. These findings may be explained by postulating a cooperativity between 5 HT1A and 5-HT2A/5-HT2C receptors with regard to the 5-HT-dependent stimulation of PRL secretion. PMID- 7495926 TI - Effects of the benzodiazepine antagonist flumazenil in PTSD. AB - OBJECTIVE: Evidence from preclinical and clinical studies suggests a role for alterations in the benzodiazepine/GABAA receptor complex in stress and anxiety. Flumazenil is a relatively pure benzodiazepine/GABAA antagonist with limited intrinsic activity. In panic disorder patients, but not healthy controls, flumazenil has been demonstrated to provoke panic attacks. METHOD: Vietnam combat veterans with PTSD (n = 14) received 90-second intravenous infusions of flumazenil 2 mg or placebo in a double-blind, crossover study design. PTSD symptomology was assessed using the PTSD Symptom Scale, and anxiety symptoms were measured with visual analogue rating scales. RESULTS: There was no significant difference in PTSD and anxiety symptoms between administration of flumazenil and placebo. CONCLUSION: Flumazenil administration does not produce an increase in anxiety and PTSD symptoms in patients with PTSD. This suggests that PTSD and panic disorder are dissimilar in terms of benzodiazepine/GABAA system function. PMID- 7495927 TI - Growth hormone response to growth hormone-releasing hormone and clonidine in depression. AB - Growth hormone (GH) responses to the alpha 2-adrenoceptor agonist clonidine and to GH-releasing hormone (GHRH) were measured in 12 patients fulfilling DSM-III-R criteria for major depressive disorder and in 12 age- and sex-matched controls. GH responses to clonidine correlated significantly with the GH responses to GHRH in the depressed patients as well as in the controls. Neither the responses to clonidine nor the responses to GHRH were significantly lower in depressed patients than in controls. Similarly, somatomedin-C (Sm-C) plasma concentrations and baseline GH concentrations were not different between the two groups. The data do not suggest that blunted GH responses to clonidine and/or GHRH represent specific features of depression. PMID- 7495928 TI - Cerebrospinal fluid corticotropin-releasing hormone, vasopressin, and oxytocin concentrations in treated patients with major depression and controls. PMID- 7495929 TI - Fluctuation of tardive dyskinesia. PMID- 7495930 TI - Effects of sleep deprivation on spontaneous eye blink rate and alpha EEG power. PMID- 7495931 TI - Glucose metabolism in murderers. PMID- 7495932 TI - Alcohol abuse and HIV. PMID- 7495933 TI - Inner-city hospitals: a call for research. AB - Inner-city hospitals serve as safety nets for poor and underserved populations. They cope with serious issues of uncompensated care, aging facilities, and inadequate reimbursement. Yet, little research exists on their needs and problems. The authors argue that research on inner-city institutions is necessary to obtain "policy" attention, legitimize demand for resources, reframe inner-city health issues as a delivery problem, and gain insights on better ways to manage inner-city hospitals. They provide data-based illustrations of future research questions related to issues of inefficiency, overstaffing, case severity, and facility renovation that can contribute to better policymaking on inner-city hospitals. The authors recognize systemic barriers to research including the unpopularity of access issues, lack of data, and absence of input from practitioners working in inner-city institutions. To remove such barriers, they suggest creation of a representative forum, original databases, increased linkage with research and policy agencies, and increased cooperation between inner-city professionals and researchers. PMID- 7495934 TI - The psychological impact of violence in underserved communities. AB - There is striking evidence that violence has a psychological impact on children and young adults in the United States, particularly those in underserved communities. Homicide is the second leading cause of death of all persons between the ages of 15 and 24 years and is the leading cause among African American youth. In 1990, more young African American men died from homicides than from all natural causes combined. Research indicates a number of factors that can predispose children to a lifetime of violence and criminal activity, including poverty, substance abuse, poor parenting skills, placement outside the home, and improper peer interaction. Evidence also indicates that early intervention through school health programs, community support systems, and, most importantly, proper parental supervision and interaction can reduce the incidence of violence and thereby the negative psychological impact violence has on children. PMID- 7495935 TI - Prenatal care use in nonmetropolitan and metropolitan America: racial/ethnic differences. AB - Pregnant women in nonmetropolitan communities are believed to use prenatal care services at lower rates than are metropolitan residents due to higher levels of poverty, lower levels of insurance coverage, and declining numbers of local hospitals and physicians. Yet scarce data exist on actual patterns of prenatal care use in nonmetropolitan areas. This study provides national estimates of prenatal care use among African American, White, and Hispanic women who delivered in 1988 in non-metropolitan and metropolitan areas of the United States. This study finds that nonmetropolitan residents are no more likely than metropolitan residents to go without care, to enter care late, or to make fewer visits. Nonmetropolitan residents are more likely to receive "inadequate" prenatal care as measured by the Kotelchuck Adequacy of Prenatal Care Utilization Index, with Hispanic residents having the highest rates of inadequate care. These findings are consistent with recent state-level reports, and they suggest the need to target prenatal care policies for populations in greatest need. PMID- 7495936 TI - De facto mental health services in the rural south. AB - Health care reform efforts highlighted the continuing scarcity of mental health services for the rural poor. Most mental health services are provided in the general medical sector, a concept first described by Regier and colleagues in 1978 as the de facto mental health service system, rather than through formal mental health specialist services. The de facto system combines specialty mental health services with general medical services such as primary care and nursing home care, ministers and counselors, self-help groups, families, and friends. The nature of the de facto system in rural areas with large minority populations remains largely unknown due to minimal available data. This article examines the availability, accessibility, and use of mental health services in the rural South and the applicability of the de facto model to rural areas. The critical need for data necessary to inform changes in health care relative to rural mental health service delivery is emphasized. PMID- 7495937 TI - The role of proton pump inhibitors in the treatment of gastro-oesophageal reflux disease. AB - Efficacy (healing, symptom relief) and cost-effectiveness are the principal reasons for the rapidly increasing use of proton pump inhibitors (PPIs) for the management of gastro-oesophageal reflux disease. EFFICACY: Mean healing rates pooled from clinical trials are as follows: on omeprazole (OME) 20 mg vs. H2 receptor antagonist. H2RA (cimetidine (CIM) 1.6 g or ranitidine (RAN) 300 mg) (eight studies) at 4 weeks, 67% vs. 37%: at 8 weeks, 81% vs. 49%: on lansoprazole (LAN) 30 mg vs. H2RA (three studies), 83% vs. 47% and 91% vs. 63% at 4 and 8 weeks, respectively. The benefit is greatest in severe disease because the H2RAs are disproportionately less effective. Heartburn is more rapidly relieved and in a higher proportion: at 4 weeks, on OME 20 mg vs. H2RA. 77% vs. 47% and on LAN 30 mg vs. H2RA, 81% vs. 46%. Both PPIs are effective in H2RA-refractory disease, approximately 80% healing occurring in 8 weeks. Relapse rates after healing vary from 25% to 85% at 6 months. Maintenance therapy sustains remissions: relapse at 1 year is, on OME 20 mg vs. RAN 300 mg (2 studies), 12% vs. 79%, and 28% vs. 55% (and 38% on OME 10 mg); on LAN 30 mg vs. 10 mg vs. RAN 600 mg, 20% vs. 31% vs. 68%. The effectiveness of the lower dose allows for dose titration. COST EFFECTIVENESS: The higher drug costs for the PPIs are offset by their higher efficacy, making their use cost effective, particularly in severe disease. Efficacy and cost effectiveness are likely to further expand the use of PPIs at the expense of H2RAs as increasing numbers of patients with milder disease are treated. PMID- 7495938 TI - The role of Helicobacter pylori in gastritis and its progression to peptic ulcer disease. AB - Helicobacter pylori infection is now recognized as the major cause of chronic gastritis throughout the world. A fraction of infected persons develop peptic ulcer disease or gastric cancer, accounting for its clinical significance. The pathophysiology of this infection can be better understood by considering five central concepts--heterogeneity of strains, persistence of infection, immunological down-regulation, physiological consequences and variability in outcome. Microbial, host and environmental factors must each contribute to the outcome variation. PMID- 7495940 TI - Unanswered questions about Helicobacter pylori. AB - There is general agreement that Helicobacter pylori eradication is indicated in all infected patients with duodenal ulcer disease and is probably indicated in all infected patients with gastric ulcer disease. However, translation of treatment recommendations into practice leads to some difficult clinical decisions. Three of the more perplexing questions are whether or not all patients with dyspepsia and H. pylori should be treated, whether or not a definitive diagnosis of ulcer should be established by an invasive method, and whether H. pylori eradication is sufficient to prevent recurrence of bleeding ulcers, especially in patient groups that have a high frequency of nonsteroidal anti inflammatory drug (NSAID) use. Another common problem is the question of whether or not to establish the success of an eradication regimen in an individual patient and the choice of method to obtain this information. There is also an obvious need to develop better antimicrobial regimens aimed specifically at Helicobacter pylori. At the basic level, almost nothing is known about the mechanisms by which H. pylori produces peptic ulcer in 10-20% of infected patients while producing gastritis in all infected subjects. There is good evidence that host factors, including intrinsic rate of acid secretion, family history and smoking are independent additive risk factors for ulcer. Ingestion of NSAIDs appears to be an independent and separate risk factor. There is evidence that strains of H. pylori that lack certain genetic markers may have a reduced likelihood of causing ulcers, but the 'ulcer' marker is present in the majority of infected subjects without ulcer.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7495939 TI - The relationship between the control of pH and healing and symptom relief in gastro-oesophageal reflux disease. AB - Gastro-oesophageal reflux disease (GERD) is generally considered to be the result of a motility disorder which permits the abnormal and prolonged exposure of the lumen of the oesophagus to the acidic gastric contents. This view is supported by experimental data, intra-oesophageal pH measurement, and the dramatic results of symptom relief and healing seen with effective antisecretory treatment. Oesophageal mucosal injury is determined by the pH of the refluxate and duration of acid exposure. Most patients experience meal-stimulated reflux during the day and the more severe cases experience 24-h acid exposure. In contrast to the H2 receptor antagonists (H2RAs), the proton pump inhibitors (PPIs) are more effective at controlling meal-stimulated acid secretion when each is given in standard doses. Therefore, the degree and duration of acid suppression throughout 24 h is greater. Treatments which maintain intra-oesophageal pH > 4 for 96% or more of the 24 h normalize acid exposure and are associated with the highest healing rates. Peptic activity is minimized at or above pH 4. The time above pH 4 is significantly longer with the PPIs than with the H2RAs. Thus, the healing-time curves for GERD (grades II-IV) are shifted to the left for the PPIs which heal a significantly greater proportion of patients earlier than the H2RAs or sucralfate. Symptoms in GERD are related to the degree and duration of oesophageal acid exposure. Symptom relief is more rapid and complete with the PPIs than with the H2RAs or other treatments in standard doses. PMID- 7495941 TI - No Helicobacter pylori, no Helicobacter pylori-associated peptic ulcer disease. AB - Virtually all duodenal ulcers (DUs) and the vast majority of gastric ulcers (GUs) are the consequence of Helicobacter pylori-associated inflammation. In DUs, the inflammation is maximal in the antrum and is associated with gastric metaplasia in the bulb. Gastrin homeostasis is disturbed by H. pylori gastritis and there is robust acid secretion. Successful eradication of the infection cures the ulcer diathesis. Amalgamated figures for ulcer relapse per year in H. pylori-positive DUs are > 60% compared with 2.6% for H. pylori-negative DU patients. The corresponding figures for GU are > 50% for H. pylori-positive and 2.0% for H. pylori-negative individuals. This striking difference in relapse rate persists, as the re-infection rate in the developed world is < 1% per year. Recurrent bleeding in bleeding-prone DUs is essentially abolished after cure of the infection. Proton pump inhibitors (PPIs) are increasingly used in eradication regimens. PPIs have intrinsic antimicrobial activity. MICs for lansoprazole (LAN) are lower than for omeprazole (OME). Two weeks of triple therapy (bismuth, tetracycline, imidazole) has, on average, a superior eradication efficacy (> or = 90%) compared with dual therapy (PPI, amoxycillin or clarithromycin) (> or = 80%). When a combination of PPI and two antibiotics has been used, results comparable to triple therapy have been reported. However, the side-effects profile and patient acceptability of PPI plus one or two antibiotic regimens are better than for traditional triple therapy.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7495942 TI - The role of acid in upper gastrointestinal haemorrhage due to ulcer and stress related mucosal damage. AB - A peptic ulcer is a lesion in which acid and pepsin are essential components of pathogenesis. Regardless of the type of patient or the setting in which the ulcer presents, the basic pathogenetic scheme is the same. The primary event is disruption of mucosal integrity. In the presence of acid and pepsin, such disruption of integrity leads to an ulcer. While rarely sufficient by itself to cause ulceration, the presence of acid is a necessary cofactor. The causes of disruption of mucosal integrity include nonsteroidal anti-inflammatory drugs (NSAIDs), Helicobacter pylori and critical illness. With the latter, tissue ischaemia may be the primary event, leading to back-diffusion of H+ ions through increased membrane permeability. Impaired mucosal buffering then leads to intramural acidosis and cell death. Risk factors for bleeding peptic ulcer in the intensive care unit (ICU) include severe trauma, sepsis, respiratory failure, and coagulopathy. Potential roles for decreasing gastric acidity in the treatment of bleeding peptic ulcer include cessation of active bleeding, prevention of rebleeding in hospital and primary prevention of bleeding. Most published studies dealing with the first two situations suggest no benefit with antisecretory therapy. However, the optimal pH for clot and platelet function may be > or = 7.0. Can such pH levels be maintained with antisecretory agents such as the proton pump inhibitors? Are the published trials adequate to demonstrate any benefit from antisecretory agents? Primary prevention of bleeding ulcer in the outpatient setting includes avoidance of NSAIDs, use of antisecretory agents and eradication of H. pylori.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7495943 TI - Drugs for pH control in upper gastrointestinal bleeding. AB - A few years ago, patients with bleeding peptic ulcers were referred to the surgeon if the bleeding did not stop. Today we have two promising new approaches to prevent emergency surgery. One is endoscopic intervention, the other is the pharmacological approach of blocking the proton pump. The endoscopical techniques of adrenaline injection fibrin-'glue' injection, polidocanol injection and heat coagulation can stop active bleeding in over 90% of cases. Pharmacologically, proton pump inhibitors can quickly achieve the optimal pH condition for support of the physiological cascade of haemostasis. The aim is to keep the intragastric pH above 6.0 for a few days. For the first time this aim can be achieved quickly and reliably by infusion of proton pump inhibitors. The optimal form of application is continuous infusion. Repeated bolus injections do not give optimal results. The optimal dosing was found to be the continuous infusion of 8 mg/h omeprazole or pantoprazole after an initial loading dose of 40-80 mg. PMID- 7495946 TI - [Family medicine, 1995: a project for everyone]. PMID- 7495945 TI - The role and limitations of H2-receptor antagonists in the treatment of gastro oesophageal reflux disease. AB - Gastro-oesophageal reflux disease (GERD) occurs in up to 44% of adults in the USA. Most individuals do not seek medical help, self-medicating with antacids. Manifestations of GERD range from symptoms without oesophagitis, which constitute the bulk of patients who self-medicate, to active oesophagitis and then to complications such as stricture and ulceration. It is the more severe cases who tend to come to the gastroenterologist, but it must be remembered that reflux symptoms are probably around 5-10 times more common than actual oesophagitis. Since acid in the refluxate is responsible for the bulk of the symptoms and mucosal damage, antacids are often used for quick relief--which of course may not be sustained. More prolonged suppression of acid secretion, such as by a histamine H2-receptor antagonist (H2RA) or a proton pump inhibitor (PPI), is required to give long-lasting symptomatic relief and heal any inflammatory change. H2-receptor antagonists inhibit acid secretion with an effect that lasts for 4-8 h with a single dose, decreasing stimulated acid secretion by around 70%. When treating oesophagitis, the H2RAs suffer from the disadvantage of their relatively short duration of action (compared with PPIs), development of tolerance, and incomplete inhibition of acid secretion in response to a meal. Therefore, it is not easy for the H2RAs to achieve optimum conditions for healing the more severe forms of oesophagitis--even very high doses may fail. In mild GERD the H2RAs have been shown to be effective in relieving symptoms.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7495944 TI - Intravenous administration of lansoprazole: a preliminary study of dose ranging and efficacy in upper gastrointestinal bleeding. AB - Using 24-h pH monitoring, we studied the optimal dose and mode of administration of lansoprazole (LAN) injection in healthy adult male volunteers in three tests of antisecretory effect. We followed this with a dose-finding study, using the drug's suppressive effect on increased gastric acid secretion due to surgical stress as the indicator. Finally, we conducted a pilot study to determine the optimal mode of administration and efficacy of the drug in patients with haemorrhage from the upper gastrointestinal tract. Twenty-four-hour intragastric pH monitoring established the order of potency of the different regimens: LAN 30 mg b.d. > LAN 15 mg b.d.; and LAN 30 mg b.d., intravenous drip infusion = LAN 30 mg b.d., bolus injection. Lansoprazole 30 mg b.d. appeared to be slightly more effective than LAN 15 mg b.d. in suppressing increased gastric acid secretion due to post-operative stress and in haemostasis in patients with upper gastrointestinal bleeding. No severe adverse effects were observed in either study. PMID- 7495947 TI - [Diabetes and pharmacoeconomics. Efficiency of insulin injection methods available in Spain]. AB - OBJECTIVES: In Spain today there are three ways of injecting insulin, the traditional syringe (TS) and two automatic methods: injector pens (IP) and preloaded syringes (PS). The main aim of the study was to compare their efficiency in normal use; and the second, to compare their effectiveness in terms of how they suited the clinical profile and needs of the users. DESIGN: A pharmacoeconomical study to minimise costs. SETTING: Two Primary Care clinics, one specialising in diabetes and the other not, and a third clinic, part of hospital out-patients. PATIENTS AND OTHER PARTICIPANTS: Systematic examination of the insulin packages and the material used by 108 diabetics (3 groups of 36, divided by the method of injection) over an average period of 51 days. MEASUREMENTS AND MAIN RESULTS: The average dose prescribed was 34.7 units (U) per day, supplied in 2.2 injections per day. The real average dose consumed was 41.4 U per day with an average daily loss of 3.1 U per injection and average re-use of needles at 7.1 times per patient. Type 11 diabetics, older patients and those with worse eye sight used TS more often. Younger and Type 1 diabetics and those who needed more injections generally used an automatic method, in particular IP. PS seemed to be used by both types of diabetic indifferently. There were significant differences found between users of TS, IP and PS regarding the doses taken (44.6, 45.1 and 34.5 U; p < 0.03), the daily loss per injection (4.5, 3.2 and 1.4 U; (p < 0.0004) and in the re-use of needles (4.1, 7.7 and 8.1 times; p < 0.02). CONCLUSION: If we suppose similar efficacy, automatic systems are more efficient in reality than traditional syringes and insulin vials. PMID- 7495948 TI - [Characteristics of the family medicine chest in the Rekaldeberri-Bilbao neighborhood]. AB - OBJECTIVE: To study the characteristics of first-aid kits used by families in the Rekaldeberri area of Bilbao (Spain) and their relationship with social and economic variables. DESIGN: Cross-sectional study. PARTICIPANTS: Households of 319 patients of the Health Center. MEASUREMENTS AND MAIN RESULTS: Interview with family and observation of first-aid kit used at home. A total number of 5170 medicines were computed with a median of 14 per household (range 1-58). More medicines were seen at households where a higher social prevailed. No relationship was found with respect to people over 65 or children under 7 nor with regard to the fact that some people were undergoing chronic treatment. The most represented anatomic & therapeutic groups are N (Central Nervous System): 19.7% and D (Dermatological): 19%. Expired medicines were found in 52.7% of the households. "Hazardous locations" were found in 3.4% of the households. CONCLUSIONS: Our population stores-up large quantities of medicines. There is a population group, however, that does not even have a basic first-aid family kit capable of meeting primary needs. Counselling relative to expiration of medicines is thus required. PMID- 7495949 TI - [Factors determining costs of drug prescriptions in primary health care]. AB - OBJECTIVE: To describe and explain the annual pharmaceutical expenditure generated by the professionals in a Health Area. DESIGN: An observational crossover study. SETTING: Primary Care. Doctors in a Health Area. PARTICIPANTS: 99 general practitioners who worked during 1992 in Area 3, Castellon. INTERVENTIONS: We used as a variable reply the pharmaceutical expenditure of each doctor during 1992, the expenditure per inhabitant and the expenditure per inhabitant standardised for the percentage of passives. We gathered as explanatory variables of the same: age, gender, whether the post was provisional or permanent, rural or urban area, MIR (resident) training or not, the model of Primary Care, frequency of attendance, whether the doctors worked there exclusively, the number of pensioners and the number of patients at work registered. MEASUREMENTS AND RESULTS: In the bivariant analysis we found a significant association of expenditure with being provisional in the post, the old model of primary care, an urban area, greater age of the doctor, greater frequency of attendance, non-exclusivity and the number of patients registered. These associations varied in line with the variable reply used. In the different models of multiple linear regression obtained, the influence of the number, non exclusivity, gender and work-place characteristics predominated. CONCLUSIONS: Greater pharmaceutical expenditure appears to be related to the number of patients registered, the absence of exclusivity, doctors being women and the features of the town where they work. PMID- 7495950 TI - [Dental caries in a school population with a program of periodical rinses with sodium fluoride]. AB - OBJECTIVE: To appraise the level of school-children's dental health after five years in operation of the buccal-dental health programme, in which children received fortnightly rinses of sodium fluoride at 0.2%. DESIGN: A crossover study. School survey of the 1993-4 academic year. PARTICIPANTS: A total of 1,674 school-children belonging to the first (6 years old), third (8), fifth (10) and eighth (13) years of EGB (basic) in the thirteen schools in this city. MEASUREMENTS AND MAIN RESULTS: Two odontologists checked the children according to the W.H.O. norms. The Index of individuals free of caries and the prevalence of caries were calculated. The first index showed its highest value at 10 (78.4%) and its lowest at 8 (53.34%). CAOD showed its lowest level at 6 years old (0.02%) and progressively rose to 13 (1.41). COD went up from 6 (0.96) to 8 (1.09) and afterwards went down till 13 years old (0.05). The restoration index was low in worn out teeth and particularly high in permanent ones (68.08%) at 13. CONCLUSIONS: The caries indexes are lower than in the majority of studies carried out in Spain, being most similar to a study carried out in the county of Mayorga de Campos (Valladolid) in 1988, where fluoridated water exists in a natural form (0.67-0.87 mg/litre), as well as to other areas in Catalonia where the same fluoride rinses were performed. PMID- 7495951 TI - [Evaluation of the completion of influenza vaccination]. AB - OBJECTIVE: To find the reasons which determine failures to comply with anti-flu vaccinations, so that these can be corrected and the coverage of this preventive action be increased. DESIGN: Observational crossover study, done by means of a telephone survey of people over 65. A questionnaire with closed questions, composed after a pilot study and validated by Cronbach's alpha. SETTING: Primary Care Centre (PCC). PATIENTS: We calculated a population sample for qualitative variables (_ = 0.05; p = 0.60; e = 0.05) of 294 people over 65, chosen from the PCC records, by means of random sampling (K = 4) stratified for age and discounting the telephone selection bias. MEASUREMENTS AND RESULTS: The proportion of vaccinated patients (60.9%) obtained in our study did not significantly differ from that in the general population. The percentage of patients included in the programme for the first time was 14%. Level of satisfaction among those vaccinated was 89.4%, with 8.9% of problems detected being light. Main causes of non-vaccination were: thinking that they didn't need it (63.5%), ignorance of the campaign (35.7%), fear of the reaction (24.3%), forgetting (10.4%). The main form of access to the campaign information was from the PCC, both through individuals and posters. Lack of information was statistically significant (p < 0.00001) as a determinant of non-vaccination, without other factors (age, sex, associated pathologies...) explaining these differences. CONCLUSIONS: Individualised and on-going health education by the PCC is fundamental. This would enable the identification of the group not vaccinated due to their express refusal and the recovery of non-vaccinated patients. PMID- 7495952 TI - [Difficulties in carrying out preventive activities in primary care]. AB - OBJECTIVE: To find the difficulties perceived by Primary Care health professionals in integrating preventive activities into their daily practice. DESIGN: A descriptive study using a survey. SETTING: Barcelones Nord and Maresme health region. PARTICIPANTS: Health professionals from primary care teams. RESULTS: The overall coverage of the survey was 47.7%. Pressure from patients or lack of time obtained the highest score (6.2), followed by the absence of a procedure or programme (4.5), absence of economic incentives (4.4) and the lack of material resources (4.2). The lowest scores corresponded to training (3.4) and motivation (2.9). CONCLUSIONS: Primary Care occupies a strategic position for carrying out preventive activities and health promotion. But in practice health professionals encounter various obstacles. Lack of time and forgetting preventive activities are basic factors. The registration sheets or other specific devices to act as reminders, together with procedures or programmes involving the joint participation of all the team members, can help resolve these problems. There is a clear need to concentrate on those problems which the professionals directly involved are most concerned with. PMID- 7495953 TI - [Adequacy of medical referrals to hospital emergency service]. AB - OBJECTIVE: To assess the correctness and diagnostic concordance in referrals of patients to hospital Casualty Departments (CD); and the possible differences between rural and urban health centres (HC) as well as between general practitioners (GP) and family and community medicine (FCM) specialists. DESIGN: Observational study of a crossover nature. SETTING: Health area 1 in the province of Badajoz. PATIENTS AND OTHER PARTICIPANTS: 800 patients, referred to their local hospital Casualty Department. MEASUREMENTS AND MAIN RESULTS: 50.5% of all referrals were considered justified. 15.1% of the patients referred were admitted to hospital. 50.7% of referrals from GPs were found to be correct, against 47% from FCM specialists; and 49% of those referred from rural HCs were correct, as against 45.9% of those referred from urban HCs (no significant differences). Only 54% of referrals had a diagnostic hypothesis, with diagnostic concordance at 50%. There was diagnostic concordance in 28.1% of GP referrals, against 33.3% of FCM specialists; and in 27.5% of those from rural HCs against 26.1% from urban HCs (no significant differences). CONCLUSIONS: We found no differences in the correctness of referrals or diagnostic concordance of patients seen in the CD of our city's health area, neither between patients referred from rural and urban HCs, nor between patients referred by GPs and FCM specialists. PMID- 7495954 TI - [Visual and auditory difficulties expressed by the aged]. AB - OBJECTIVE: To determine the proportion of the elderly who have difficulties in their perceptive functions (seeing and hearing) and associated factors in the area of health self-perception and functional capacity. DESIGN: Observational and crossover, using an interview. SETTING: Community. PARTICIPANTS: 787 elderly people not in institutions; 93.8% reply rate. MEASUREMENTS AND MAIN RESULTS: Using a questionnaire, data were obtained on deficiencies in their sense organs and self-perceived health. Validated scales to determine functional capacity (physical, psychic and social) were used: Katz, Lawton and Brody indexes, Cognitive mini-test, DUKE-UNC questionnaire and Geriatric Depression scale. A serious difficulty or inability to read or watch television was expressed in 17.3% of cases (CI 95%; 14.8-20.0) and to follow a normal conversation in 10.5% (CI 95%; 8.4-12.6). The proportion of the elderly dependent on others to carry out their normal daily activities was 2.2 times greater among those who displayed severe visual problems (CI 95%; 1.7-2.9) and 1.8 times greater if they had hearing difficulties (C.I. 95%; 1.4-2.3). There was a linear tendency between greater visual or auditory problems and less social support (p = 0.01), greater dependency on others for basic daily activities (p < 0.001) and worse perceived health (p < 0.001). CONCLUSIONS: The elderly, especially the oldest among them, frequently display hearing and visual problems, which are related to low self perception of health and greater functional incapacity. PMID- 7495955 TI - [Screening for cancer]. PMID- 7495956 TI - [Drug prescriptions in general medicine clinics or drugs prescribed by primary care physicians?]. PMID- 7495957 TI - [Clinical usefulness of drug surveillance studies]. PMID- 7495958 TI - [Teaching of family medicine: the figure of the hospital tutor]. PMID- 7495959 TI - Virus validation procedures: practical aspects. AB - The Ad Hoc Working Party on Biotechnology and Pharmacy of the Committee for Proprietary Medicinal Products has developed a number of guidance notes for manufacturers intending to submit applications for market authorization in the European Union which includes one dealing specifically with the validation of production processes for the removal or inactivation of viruses. The strategies used to minimize the risk of viral transmission by biological products are: screening the source materials for viral contamination; examining the ability of the production process to remove or inactivate viruses; and examining the final product for evidence of viral contamination. It is concluded that validation studies provide a significant assurance of viral safety when properly carried out, using appropriate relevant and model viruses. PMID- 7495960 TI - Virus inactivation procedures: an overview. AB - Viruses can easily be transmitted by intravenous infusion of contaminated blood or blood products. Reduction of viral transmission is achieved by suitable donor selection, screening of donations, and choice of manufacturing processes. The components of this strategy should not be considered individually but are complementary. An overview is presented of the combined strategy for reducing the likelihood of virus transmission via blood and blood products. PMID- 7495962 TI - Hepatitis A virus and blood products: virus validation studies. AB - As there have been reports of the transmission of hepatitis A virus by single units of blood, virus validation studies were carried out to determine the fate of the virus at four major steps (cryoprecipitation, solvent/detergent inactivation, DEAE chromatography and lyophilization) involved in the production of solvent-detergent inactivated factor VIII products. If the reduction in infectivity is additive over the four steps, the total reduction would be between 8.6 and 9.3 log10. It is unlikely that such a reduction would provide a sufficient safety margin, particularly if the original plasma pool from which the FVIII was prepared, was obtained from a population in which hepatitis A virus was not rare. In such a situation, an additional method of effectively reducing hepatitis A virus infectivity should preferably be included in the production process. PMID- 7495961 TI - Good manufacturing practice and viral safety. AB - The concept of virus inactivation during the manufacture of blood products raises questions about possible recontamination of the product by the environment. A strict regime of good manufacturing practice (GMP) is mandatory. The guidelines originally issued by the World Health Organization (WHO), and now law in most countries, are an excellent basis for the operation of a production plant. The following elements of GMP require special concern: (i) All functions shall be defined in a clear organization chart. (ii) Personnel shall be appropriately trained for the job and to perfect hygiene. (iii) Buildings and facilities, as well as supply systems, shall exclude the possibility of recontamination of already virus-inactivated materials. (iv) Equipment shall be easy to clean and fully sterilizable. (v) Production shall follow appropriate written procedures. (vi) The Quality Control Organization shall monitor the process by in-process controls and review the records for possible deviations. All GMP issues are coordinated by a Quality Assurance Organization that also reviews the overall performance of the operation. The maintenance of viral safety of the products basically depends upon the full commitment of all bodies involved to proper and non-negotiable GMP. PMID- 7495963 TI - Self-sufficiency and blood transmitted diseases. AB - The Council of Europe and the EEC Council of Ministers have strongly promoted self-sufficiency for plasma products on the basis of voluntary non-remunerated donors. Several European countries have a programme of self-sufficiency with plasma products, either with national fractionation plants (e.g. Belgium, Finland) or based on contract fractionation (e.g. Norway, Slovenia). Advantages of national self-sufficiency includes epidemiological factors, economical factors and also ethical and moral issues. Self-sufficiency is one of the basic conditions for reducing the hazard of transmission of infectious diseases. Norway has been self-sufficient with coagulation factors since 1981. Price mechanisms and market forces have been important factors in ensuring the necessary plasma volume, and fractionation methods rendering high yields of factor VIII are initially preferred. This policy has resulted in a low prevalence of antibodies against human immunodeficiency virus (6%), hepatitis B virus (28%) and hepatitis C virus (41%). No Norwegian haemophiliacs have been infected with hepatitis A through FVIII concentrates. PMID- 7495964 TI - Hepatitis A virus and polymerase chain reaction amplification: methodology and results. AB - Hepatitis A infection among patients receiving solvent/detergent inactivated factor VIII preparations in various locations in Europe have been documented recently. In investigations in Italy, Germany and Ireland, polymerase chain reaction (PCR) amplification was used to detect hepatitis A virus in frozen plasma pools, purified factor VIII, patient sera and samples from animal transmission studies; nucleic acid sequencing was used to clarify and identify the virus responsible based upon genotype analysis. Unique virus strains were found among the cases in Italy and Germany, and identical virus sequences were also found in some factor VIII lots. However, with the exception of the Italian investigation, lack of appropriate samples have precluded the identification of virus in these outbreaks. In addition, animal infectivity studies have not been successful in demonstrating infectivity under laboratory conditions. We discuss the limitations of PCR amplification with respect to detecting virus within these situations, and the necessity for the corresponding epidemiologic investigations. PMID- 7495965 TI - Detection of infectious hepatitis A virus in blood factor concentrates by experimental infection of the New World primate Saguinus fuscicollis. AB - At present, the hepatitis A virus cannot be detected readily by cell culture techniques. However, the New World primate Saguinus fuscicolis is particularly susceptible to this virus. This paper gives details of the in vivo detection of hepatitis A virus in S. fuscicolis, and the method is used to show that a blood factor VIII preparation, suspected of being contaminated with hepatitis A, did not contain the virus. PMID- 7495966 TI - Approaching the ideal: therapeutic requirements for factor VIII concentrates. AB - The requirements that a factor VIII concentrate should fulfil, as adopted by the Belgian Health Council, are reviewed. Lack of immunogenicity is an important issue; the Belgian experience suggests that a prospective randomized trial allows rather rapid detection of grossly increased immunogenicity. An alternative is provided by prospective cohort studies, using a uniform protocol to evaluate the behavior of each product, and an independent data monitoring centre. Residual infectivity is illustrated by an outbreak of hepatitis A in heavily treated patients. A prospective randomized trial in Belgium is planned to evaluate the benefits and risks of a double-virus inactivated product. PMID- 7495967 TI - Development and biochemical characterization of a double-virus-inactivated factor VIII preparation. AB - This paper describes the development of a double-virus-inactivated preparation. The original goal was to inactivate and/or remove parvovirus from plasma-derived factor VIII. More recently, attention has also been focused on the controversial issue of transmission of the non-lipid-coated hepatitis A virus by preparations of factor VIII which have been solvent/detergent-treated and purified by ion exchange chromatography. PMID- 7495968 TI - Virus validation experiments on the production process of OCTAVI SDPlus. AB - The inactivation of both transfusion-relevant and model viruses by modified pasteurization has been evaluated following the established guidelines of the European Union Committee for Proprietary Medical Products Ad Hoc Working Party on Biotechnology/Pharmacy. This heat treatment in solution for 10 h at 63 degrees C was introduced into the manufacturing process of OCTAVI, a very high purity factor VIII concentrate stabilized by von Willebrand factor. It could be demonstrated that both enveloped (human immunodeficiency virus, herpes simplex virus, pseudorabies virus) and non-enveloped viruses (poliovirus, coxsackievirus, hepatitis A virus) were inactivated by this heating step with an efficacy of greater than 4.5 log10 TCID50. The combination of the solvent/detergent step already used in the manufacture with this modified pasteurization leads to a double virus-inactivated factor VIII concentrate (OCTAVI SDPlus) with a viral safety distinctly superior to monoinactivated products. PMID- 7495969 TI - Viral iatrogenic contamination and safety of biologicals. AB - The use of biologicals such as bacterial or viral vaccines have an inert risk as a result of their preparation. In the last century it was difficult to determine the correlation of the clinical accidents until the characteristic viral genome became available. Generally, the frequency of accidents provoked by viral vaccines has diminished due to (1) the improvements of the methods used for viral inactivation and purification, (2) the introduction of good manufacturing practices and rigorous regulatory policy and (3) the progress in the post clinical surveillance of adverse reactions after vaccine administration, allowing a better risk-benefit analysis. PMID- 7495970 TI - Inhibitor development in haemophiliacs: theoretical background and clinical aspects. AB - The development of factor VIII inhibitors is one of the most serious complications of repeated transfusions in haemophilia patients, but it is not clear why the complication develops only in some haemophiliacs. This paper discusses the nature of FVIII inhibitors, their occurrence and detection, and outlines treatment procedures for haemophilia patients who show FVIII inhibitors. PMID- 7495971 TI - Strategy for pre-clinical evaluation of factor VIII concentrates. AB - A series of methods and assay systems was designed, using both mouse monoclonal antibodies and purified human polyclonal antibodies, by which alterations in the antigenic properties, and potentially therefore in the immunogenic properties, of FVIII concentrates could be identified. Those methods could be applied to the pre clinical evaluation of FVIII concentrates. It has become evident that very subtle alterations in FVIII can have dramatic effects on its antigenicity and immunogenicity and it is suggested that an analysis of FVIII preparations, using reagents such as mouse monoclonal antibodies, should be carried out as a pre clinical evaluation. It is also thought that any haemophiliac patient is at risk of developing anti-FVIII antibodies, even though he had been considered as tolerant to the infusion of FVIII. The diversity of FVIII concentrates now available on the market has multiplied the chances of encountering a product that would give an immune response in some patients and this further stresses the need for a careful check of FVIII concentrates. PMID- 7495972 TI - Competitive ELISA to evaluate the immunogenicity of double-virus-inactivated factor VIII in rabbits. AB - Viral inactivation is a critical step during the manufacture of factor VIII formulations for use in humans. Indeed, viral inactivation procedures may alter the three-dimensional structure of factor VIII resulting in the formation of new epitopes which, in turn, may lead to the synthesis of inhibiting antibodies, and hence to a decreased therapeutic activity. A rabbit model was used to compare the immunogenicity of a solvent/detergent (SD)-inactivated formulation with a double inactivated (SD plus heating in solution, SDP) formulation of factor VIII. Two groups of five rabbits were immunized by six s.c. injections of each formulation. The serum of each animal was incubated with various amounts of the competing antigens, e.g. factor VIII SD or factor VIII SDP. The remaining free polyclonal antibodies were assayed by ELISA. Curves obtained with both antigens were compared for each serum. Both factor VIII SD and factor VIII SDP decreased the amount of antibodies raised to either formulation in a dose-dependent manner without observable differences. These results suggest that no new epitopes were present on factor VIII SDP as compared with factor VIII SDP as compared with factor VIII SD and that no epitope deletions occurred, supporting the view that the double-inactivation procedure does not change the immunogenicity of factor VIII SD. This model is proposed as a tool to detect changes in the immunogenicity of proteins which may be modified. PMID- 7495973 TI - Factor VIII concentrates and the immune system--laboratory investigations. AB - Evidence suggests that haemophiliacs treated with factor VIII concentrates show abnormalities in immune functions. The basis of this is not clear, but some factor VIII concentrates down-regulate Fc receptors on monocytes which may explain the impaired function of these cells. Some concentrates inhibit lymphocyte proliferation and interleukin-2 secretion by human T-cell lines and peripheral blood lymphocytes. They can also inhibit activity of other cytokines such as interleukin-4 and interleukin-5 and secretion of cytokines such as interleukin-1 and granulocyte macrophage colony stimulating factor. These effects are product-related and vary from total inhibition to virtually no detectable inhibition. Of particular significance is that the degree of inhibition is not related to the purity or gross protein composition of the products. The inhibitory activity is not due to factor VIII itself as antibody affinity purified factor VIII products are entirely non-inhibitory. The main inhibitory protein components appear to be of approximately 200 kDa and 60 kDa (by gel filtration). Recent evidence suggests that transforming growth factor-beta (TGF beta), derived from the plasma used for fractionation, is a major contaminant of 'inhibitory' concentrates and is responsible for the effects, observed in vitro, of concentrates on cytokine secretion or activity. The levels of TGF-beta varied between products and correlated with inhibition of interleukin-2 secretion from stimulated T-cells. The presence of TGF-beta in concentrates may therefore explain the immunosuppression observed in recipients of these products. Correlation of the inhibitory effects with clinically important consequences such as increased susceptibility to infections or decreased CD4 counts also remains to be established. PMID- 7495974 TI - High purity factor VIII concentrates for the treatment of HIV-positive patients with haemophilia. AB - Some factor VIII concentrates cause downregulation of the immune system in HIV positive haemophilia patients. This effect appears to be related to the purity of the concentrate, but not to its method of preparation. The optimum purity for preventing downregulation is not known at present. PMID- 7495976 TI - Preliminary clinical experience: pharmacokinetics and tolerance of a double-virus inactivated factor VIII preparation. AB - The incorporation of viral inactivation steps into the manufacture of factor VIII concentrates has markedly improved the safety margin of the product, but in order that viruses, such as hepatitis A, are not transmitted by infusion of factor VIII preparation, some manufacturers include a further process step, such as heat inactivation. There is the possibility that such a step will change the final factor VIII product and it was investigated whether there are any pharmacokinetic differences between the products Octavi S/D and Octavi S/D+heat treatment. It is concluded that Octavi S/D+heat treatment can be given in doses equal to Octavi S/D or other factor VIII concentrates. PMID- 7495975 TI - Prospective clinical trial of high-purity factor VIII preparations in haemophiliacs. AB - An assessment of the immune status of both HIV-positive and HIV-negative haemophiliacs showed hypo-responsive and anergic patients in both groups. In HIV positive patients, the immune suppression, in addition to HIV itself, may be caused by other viral infections such as all types of hepatitis as well as by factor VIII concentrates. The introduction of new technologies for preparing factor VIII concentrates led to the question of whether higher purity is associated with a protective effect on the immune system, which might improve the clinical course of HIV-positive patients. Factor VIII from two manufacturing processes was investigated, one prepared using an ion-exchange S/D step, and the other utilizing a monoclonal antibody step. It is concluded that changing the product, from the monoclonal antibody to the ion-exchange and vice versa in the HIV-negative patients does not cause a change in the immune profiles. PMID- 7495978 TI - Introduction: basic science aspects of breast cancer. PMID- 7495977 TI - Clinical evaluation of virus safety and inhibitor incidence: statistical considerations. AB - The basic statistical issue in pharmacovigilance is to claim, with reasonable certainty, that the incidence of an event of interest in a population of subjects is less than a certain value. How many subjects and events must be observed before such a claim can be made? A first situation of practical importance is when a product has been on the market for some period of time, and the safety of this product regarding some outcome of interest is questioned (for instance, the viral safety of blood products). Having observed a few occurrences of the event of interest, how confident can we be that the product is responsible for an elevation of the incidence of this event compared with the baseline incidence in a reference population? This issue will increasingly need to be addressed prospectively: how many subjects need to be treated and how many events observed, to be reasonably certain that a product is safe? Multi-stage designs are appropriate to address this question, yet they do not seem popular in pharmacovigilance. Such approaches could complement the standard recommendations to assess the safety of blood products, which are adequate as a first screen against major safety problems, but wholly inadequate for the long-term surveillance of subjects at risk of rare events. It will be argued that, from a regulatory perspective, the implementation of prospective protocols for pharmacovigilance, using appropriate statistical tools, would permit a tight control of the safety of new products, while making these products available as early as possible to the patients who need them. PMID- 7495979 TI - Tumor suppressor genes and their alterations in breast cancer. AB - Tumor suppressor genes were first identified in inherited cancer syndromes, and were later shown to be involved in sporadic cancer as well. Over a dozen tumor suppressor genes have now been discovered including a breast and ovarian cancer susceptibility gene, BRCA1. Germ-line mutation of tumor suppressor genes is associated with an inherited predisposition to a limited tumor spectrum, but somatic mutations in tumors are heterogeneous. The 16 characterized tumor suppressor genes are involved in diverse functions including cell cycle regulation, check point control, transcriptional repression, signal transduction modulation, and DNA repair. These studies demonstrate that disturbance in cell proliferation, genetic stability and cell death could all precede tumorigenesis. PMID- 7495980 TI - Oncogene activation and oncogene cooperation in MMTV-induced mouse mammary cancer. AB - In this short review, we will give an overview of the various genes that can be activated by insertion of proviral DNA of the mouse mammary tumor virus during the formation of mammary cancer. These genes fall within three families: Wnt genes, FGF genes and Notch-related genes. We will summarize our current understanding of the roles of these genes in tumorigenesis and in normal development, and the mechanisms of action of their gene products. Finally, we will give some examples of cooperation between these genes in various biological settings. PMID- 7495981 TI - Interaction between erbB-receptors and heregulin in breast cancer tumor progression and drug resistance. AB - The type I growth factor receptor family is increasingly recognized as important in the development and maintenance of breast cancer. The family currently consists of four closely related members: the epidermal growth factor receptor (EGF-R/erbB-1), erbB-2, erbB-3 and erbB-4. Putative ligands which bind directly to or indirectly activate erbB-2/3/4 have been characterized recently. This still growing family of EGF-related growth factors includes gp30, its homolog heregulin (HRG), the rat homolog neu differentiation factor (NDF), glial growth factors (GLIA), ARIA and a 50 kDa factor from COLO 16 cells. The understanding of the function, biology and interactions of these growth factor receptors and their ligands will have far-reaching implications for the prognosis and treatment of breast cancer. This review focuses on advances and future directions for further investigations intended to clarify the mechanism and significance of erbB/ligand interactions in breast cancer. PMID- 7495982 TI - Rodent models for the study of etiology, prevention and treatment of breast cancer. AB - Rodent models have been, and continue to be, important in both defining the etiology of breast cancer, and in generating new prevention and treatment strategies directed at this prevalent malignancy. Of all solid cancers, breast cancer has the widest variety of rodent models. Models in rats and mice include both spontaneous, induced and transplantable carcinomas in standard strains. In addition, many strains are available for modeling inherited susceptibility and resistance to breast cancer. Finally, many new models that have taken advantage of techniques from biotechnology such as transgenic, knock-out and gene therapy technologies are being used to evaluate new prevention and treatment strategies for breast cancer. PMID- 7495983 TI - Histopathology of transgenic mouse mammary tumors (a short atlas). AB - Mammary hyperplasias, dysplasias and tumors have been described in many strains of transgenic mice. Many of the transgenes produce characteristic disturbances of growth, development and neoplasia. The disturbances can now be classified into groups. A classification of transgenic tumors and an atlas illustrating some characteristic examples of common morphological changes in transgenic mammary tissues is provided. PMID- 7495984 TI - Mammary gland tumor formation in transgenic mice overexpressing stromelysin-1. AB - An intact basement membrane (BM) is essential for the proper function, differentiation and morphology of many epithelial cells. The disruption or loss of this BM occurs during normal development as well as in the disease state. To examine the importance of BM during mammary gland development in vivo, we generated transgenic mice that inappropriately express autoactivating isoforms of the matrix metalloproteinase stromelysin-1. The mammary glands from these mice are both functionally and morphologically altered throughout development. We have now documented a dramatic incidence of breast tumors in several independent lines of these mice. These data suggest that overexpression of stromelysin-1 and disruption of the BM may be a key step in the multi-step process of breast cancer. PMID- 7495985 TI - Involvement of extracellular matrix constituents in breast cancer. AB - It has recently been established that the extracellular matrix is required for normal functional differentiation of mammary epithelia not only in culture, but also in vivo. The mechanisms by which extracellular matrix affects differentiation, as well as the nature of extracellular matrix constituents which have major impacts on mammary gland function, have only now begun to be dissected. The intricate variety of extracellular matrix-mediated events and the remarkable degree of plasticity of extracellular matrix structure and composition at virtually all times during ontogeny, make such studies difficult. Similarly, during carcinogenesis, the extracellular matrix undergoes gross alterations, the consequences of which are not yet precisely understood. Nevertheless, an increasing amount of data suggests that the extracellular matrix and extracellular matrix-receptors might participate in the control of most, if not all, of the successive stages of breast tumors, from appearance to progression and metastasis. PMID- 7495986 TI - The development of a functionally relevant cell culture model of progressive human breast cancer. AB - Normal mammary homeostasis, and by implication tumorigenesis, are dependent upon the dynamic interplay between epithelial cells, stromal components and the extracellular matrix. To study the evolution of human breast cancer, a functionally relevant cell culture model is required which recognizes the complexity of the mammary gland's microenvironment. The development of an appropriate breast epithelial cancer cell model will be dependent on the ability to recreate the 'normal' and 'neoplastic' tissue microenvironment in culture. Towards this goal, a 3-dimensional extracellular matrix (ECM) assay, employing a reconstituted basement membrane, has been developed which allows for the rapid and accurate discrimination of normal and neoplastic cells when cultured. To investigate stromal/epithelial cell interactions, we have developed a tumor environment assay which essentially mirrors the tumor microenvironment histologically. The use of a novel, near diploid, human breast epithelial cell line, HMT-3522, which has transformed spontaneously with passage in culture, together with these 3-dimensional culture assays is expected to provide meaningful markers of initiation and progression. PMID- 7495987 TI - Preneoplastic transformation of human mammary epithelial cells. AB - Recent in-vitro models of mammary epithelial cell (MEC) immortalization have provided a practical approach to begin to dissect the molecular mechanisms of breast tumorigenesis. Introduction of a single oncogene, the human papilloma virus (HPV)-16 E6, induces efficient and reproducible preneoplastic transformation of normal MECs, by inducing degradation of the tumor suppressor protein p53. The role of p53 has also been demonstrated by analyses of a model of gamma-radiation-induced MEC transformation. Recently, efficient retroviral gene transfer has allowed identification of multiple mammary epithelial cell types that show distinct susceptibilities to HPV E6 and E7 oncogenes, indicating a cell type-specific predominance of the tumor suppressor proteins p53 and Rb which are targeted by E6 and E7, respectively. Further analyses of these models are likely to elucidate the biochemical mechanisms of early mammary tumorigenesis. PMID- 7495988 TI - Alternative/complementary therapies used by persons with HIV disease. AB - The authors examined the kind of alternative and complementary therapies used by persons with HIV disease. A convenience sample of 145 persons with HIV disease who were receiving health care in either New York City (n = 72) or Boston (n = 73) completed the Alternative Therapies Check List. This instrument consists of 55 alternative or complementary therapies; respondents were asked to identify whether they knew the therapy and how often they used it. Frequencies were computed, demonstrating that all of the identified alternative or complementary therapies were used by at least one person with HIV disease. The different therapies were ranked according to their mean or average use. Findings showed that vitamins were the most commonly used and trigger was the least commonly used alternative/complementary therapy. PMID- 7495989 TI - New perspectives on HIV pathogenesis. Viral replication is continuous throughout the course of disease. PMID- 7495990 TI - Opioid use in HIV patients with neurological changes. AB - The neurological changes that are common in the HIV population may complicate the use of analgesics, particularly opioids. These changes, in combination with care providers' fear of opioids, often lead to the inappropriate use of antagonists, such as naloxone. Used injudiciously, naloxone can lead to withdrawal syndrome, return of severe pain, and other adverse effects. The authors describe the many disease and treatment-related causes for pain in patients with HIV disease, as well as common neurologic conditions that alter cognition and complicate the use of opioids. A case study is included to illustrate the dangers of inappropriate naloxone use. PMID- 7495991 TI - HIV/AIDS education for nurses in Poland. AB - Funded by an agreement with the World AIDS Foundation, a team of American and Polish healthcare professional was established to provide expert and timely training on HIV/AIDS course for nurses was conducted in Gdansk, Poland. The course identified significant professionals concerns of Polish nurses, including fear of infection, discomfort with male sexuality, and lack of information about drug users. Precourse and postcourse evaluations demonstrated significant changes in level of comfort during discussions of sensitive topics and improvement in instructional self-confidence. PMID- 7495992 TI - Treatment review. Testosterone (Testoderm). PMID- 7495993 TI - Public health under threat. PMID- 7495994 TI - Management of the hematological manifestations of HIV infection and AIDS. AB - Hematological abnormalities, characterized by cytopenias, are a problem commonly encountered by persons with HIV or AIDS. Cytopenias can be multifactorial in their causation and potentially life threatening to this population. An understanding of the various causes of these cytopenias and the appropriate nursing care are tantamount to nursing practice and to ensure quality of life. Goals of medical and nursing care are directed at assessment, interventions, and patient education to prevent or minimize the complications of hematological abnormalities. This article reviews current knowledge regarding the causes, pathophysiology, treatments, and nursing care of HIV/AIDS-related hematological abnormalities. PMID- 7495995 TI - Assessment of tuberculosis: is 'rapid assessment' needed? PMID- 7495996 TI - In vitro antimycobacterial activity of a new quinolone, levofloxacin (DR-3355). AB - OBJECTIVE: To examine in vitro antimycobacterial activity of levofloxacin. DESIGN: Minimum inhibitary concentrations (MICs) of levofloxacin for various mycobacterial species were determined by the agar dilution method using 7H11 medium and compared with those of ofloxacin. Antimicrobial activity of levofloxacin against Mycobacterium tuberculosis and M. intracellulare phagocytosed in murine peritoneal macrophages was measured in terms of reducing cell-associated bacterial colony forming units (CFUs). RESULTS: MICs of levofloxacin against M. tuberculosis, M. kansasii, M. marinum, M. scrofulaceum, M. avium, M. intracellulare, M. fortuitum, and M. chelonae were 2 to 4 times lower than those of ofloxacin. Levofloxacin exhibited higher efficacy in reducing bacterial CFUs in macrophages than ofloxacin. CONCLUSION: Levofloxacin possessed more potent in vitro antimycobacterial activities as compared to that of ofloxacin. PMID- 7495997 TI - Diagnosis of tuberculous meningitis: a comparative analysis of 3 immunoassays, an immune complex assay and the polymerase chain reaction. AB - OBJECTIVE: To compare 3 immunoassays, an immune complex assay, and an application of the polymerase chain reaction (PCR) for the diagnosis of tuberculous meningitis (TBM). MATERIAL: Cerebrospinal fluid (CSF) from 33 patients with TBM and from 34 control patients with infectious and non-infectious CNS diseases was analysed. RESULTS: The antibody immunoassays were either nonspecific or insensitive. However, detection of mycobacterial IgG immune complexes correlated strongly with infection, as they were detected in the CSF from 64% of the patients with TBM compared to only 3 (9%) of the control samples. PCR analysis, using Mycobacterium tuberculosis-specific oligonucleotide primers, also strongly correlated with infection, as DNA was amplified from 54% of the samples from patients with TBM, but from only 2 (6%) of the control samples. Both 'false positive' samples were also positive in the immune complex assay and came from 2 patients with otogenic brain abscesses. It is conceivable that these patients suffered from otogenic tuberculosis with secondary non-mycobacterial meningitis. When combining the immune complex assay with DNA-detection by PCR, 100% of the culture positive and 74% of culture negative samples were found to be positive, while maintaining a high specificity. CONCLUSION: Parallel analysis to detect mycobacterial immune complexes and M. tuberculosis-specific DNA by PCR from the CSF of patients may offer a sensitive and specific tool for the diagnosis of TBM. PMID- 7495998 TI - The epidemiology of tuberculosis in Gran Canaria, Canary Islands, 1988-92: effectiveness of control measures. AB - SETTING: Since there is no national programme for tuberculosis control in Spain and the epidemiological situation in this country is unknown, a programme against tuberculosis in Gran Canaria (Canary Islands) was started in 1987. OBJECTIVE: To analyze the impact of this control programme, particularly the trend of new cases of tuberculosis by age groups and risk factors for 1988-92. DESIGN: A detailed search of all new cases of tuberculosis was carried out by examining the files of all public and private hospitals and outpatient clinics of the island, the records of the Services of Clinical Microbiology and Pathology of the public hospitals, and the registers of patients with AIDS. RESULTS: The annual rates of new cases of tuberculosis per 100,000 were 32.2 in 1988 (214 cases), 26.2 in 1990 (174 cases), and 29.4 in 1992 (196 cases), and the rates of sputum-positive cases 13.3 (89 cases), 12 (80 cases) and 15 (100 cases), respectively. The distribution of tuberculosis cases by age peaked in the age groups 40-49 and 30-39 years. Between 1988 and 1992, statistically significant differences were found in the percentage of cured patients (21% [45/214] vs 85.7% [168/196]), patients with a delay in diagnosis > 3 months (40.2% [86/214] vs 21.9% [43/196]), and contacts evaluated (45.3% [97/214] vs 90.3% [177/196]). However, an upward trend in the incidence of tuberculosis for 1990-92 was found in association with an increase of new cases among the homeless, immigrants, and HIV-infected people. When these three population groups were excluded from the analysis, the annual rate for the total population showed a progressive decline from 1988-92. CONCLUSIONS: The data studied here show that successful priority control measures (cure rate, early diagnosis, study of contacts) have been achieved in this island. It is necessary, however, to develop strategies for combating the tuberculosis problem in specific high risk populations. PMID- 7495999 TI - Isoniazid preventive therapy for pulmonary tuberculosis sequelae: which patients up to which age? AB - SETTING: Preventive therapy with isoniazid (INH) is usually recommended for all patients with pulmonary fibrotic lesions compatible with previous tuberculosis (TB). OBJECTIVE: To determine the optimal strategy between a 6- to 12-month course of prophylactic INH and therapeutic abstention in different age groups and in patients with severe coexisting diseases that limit life expectancy, such as chronic heart failure (CHF) or chronic obstructive pulmonary disease (COPD). DESIGN: A Markov decision analysis model that balances the risk of developing active TB against TB-related mortality, the risk of INH-related hepatitis, and INH-related death. RESULTS: In all groups of patients, prophylactic INH clearly reduced the number of deaths from TB, whereas very few cases died from INH related toxicity. However, because INH-related deaths occur early, and TB-related deaths occur early or late, the gain in life expectancy was small. Particularly for patients with short survival such as those older than 80 years and those with CHF or COPD, the average gain in life expectancy provided by prophylactic INH did not exceed one week. CONCLUSIONS: Our analysis confirms that prophylactic INH is beneficial to all cohort groups of patients. However, in the case of very old age or severe disease, the gain in life expectancy is minimal. In these cases, the decision to give INH prophylaxis should be tailored on an individual basis with special consideration given to the patient and his environment. PMID- 7496000 TI - Clinical features of HIV seropositive and HIV seronegative patients with tuberculous lymphadenitis in Dar es Salaam. AB - SETTING: The medical wards of a referral hospital in Dar es Salaam, Tanzania. OBJECTIVE: To investigate the impact of HIV infection on clinical features in tuberculous lymphadenitis. DESIGN: A prospective clinical study of HIV seropositive and HIV seronegative patients with lymphadenopathy. RESULTS: Of 128 patients with peripheral lymphadenopathy, 24 had no tuberculosis (TB) and in 10 patients TB was found only in other organs. The remaining 94 patients, of whom 76% were HIV seropositive, formed our study population. TB lymphadenitis was considered proven in 89 and probable in 5 patients. Disseminated TB (both TB adenitis and TB in other organs) was diagnosed more often in HIV seropositive than in HIV seronegative patients (52% versus 26%, P < 0.03). 59% of the 71 HIV infected patients compared to only 4% of the 23 patients without HIV infection were over 30 years of age (P < 0.02). The following clinical features were significantly associated with HIV infection: dyspnoea, respiratory rate > 20/min, low motility score (bedridden), neurological abnormalities, hepatomegaly, splenomegaly, lymph node size < 2.5 cm, negative PPD skin test, lymphopenia (< 1000/cm3) and presence of pleural fluid. CONCLUSION: Co-infection with HIV influences several clinical and laboratory features in patients with tuberculous lymphadenitis. PMID- 7496001 TI - [Clinical trial of a combination of three drugs in fixed proportions in the treatment of tuberculosis. Groupe de Travail sur la Chimiotherapie de la Tuberculose]. AB - SETTING: The Matiben Chest Clinic at the West Algiers University Teaching Hospital, and 3 outpatient clinics specializing in tuberculosis and lung disease in Algiers. OBJECTIVE: To determine the tolerance and efficacy of a fixed proportion combination of 3 antituberculosis drugs (per tablet: 50 mg isoniazid + 120 mg rifampicin + 300 mg pyrazinamide) given during the first 2 months of a daily 6-month chemotherapy regimen. DESIGN: Random prospective treatment trial comparing a group of 124 patients receiving the triple combination with another group of 126 patients receiving the 3 drugs separately during the initial treatment phase. The continuation phase was identical for the 2 groups. Comparison of tolerance in the first 2 months, and of the failure and relapse rates (respectively at the end of treatment and 24 months after the end of treatment). RESULTS: During the first 2 months side-effects were significantly more common in the group receiving the drugs separately. At the end of treatment and during the following 24 months there were no significant differences in the cumulative rates of observed failures and relapses (2% and 1%). CONCLUSION: The triple combination studied could replace the separate drugs in the initial treatment phase in countries where the bioavailability of the drugs used has been proven. PMID- 7496002 TI - An audit of HIV testing and HIV serostatus in tuberculosis patients, Blantyre, Malawi. AB - SETTING: Queen Elizabeth Central Hospital, Blantyre, Malawi. OBJECTIVE: An audit of voluntary HIV testing, with pre- and post-test counselling, of adult patients diagnosed with all types of tuberculosis. DESIGN: A review of case files of adult patients with tuberculosis registered with the District Tuberculosis Officer, Blantyre, between April 1993 and March 1994. RESULTS: There were 1095 tuberculosis patients, mean age 32 years, of whom 665 (60.7%) had HIV-serological testing. 496 patients (74.6% of those tested) were HIV seropositive. 73% of patients who were hospitalized for the initial intensive phase of treatment were HIV-tested compared with 37% of patients who received ambulatory chemotherapy (P < 0.001). In patients HIV-tested, 5 did not wish to know their results and post test counselling was done in 516 (78%). 23 patients refused HIV testing. 362 (84%) patients not HIV-tested never received pre-test counselling. Of 664 patients who received 2SRHZ/6HT(E) in hospital, 84 (12.6%) patients died and 8 (1.2%) absconded. The abscondee rate was unrelated to HIV serostatus. CONCLUSION: A large proportion of tuberculosis patients who receive supervised treatment in hospital accept confidential HIV testing and the abscondee rate is low. The clinical management of patients is improved. PMID- 7496003 TI - Diagnostic utility of the estimation of mycobacterial Antigen A60 specific immunoglobulins IgM, IgA and IgG in the sera of cases of adult human tuberculosis. AB - SETTING: An ELISA assay based on mycobacterial Antigen A60 for the estimation of Mycobacterium-specific immunoglobulins in the serum has been used successfully for the rapid diagnosis of tuberculosis in studies done in the Western countries. There are hardly any similar large scale studies in India. OBJECTIVE: To evaluate the utility of this ELISA test for rapid diagnosis of different clinical forms of tuberculosis in an adult Indian population. DESIGN: ELISA test based on mycobacterial antigen A60 (Anda Biologicals, France) was used to estimate specific IgM, IgA and IgG antibodies in the sera obtained from 337 cases of tuberculosis and 131 controls in the population of Delhi (India). RESULTS: Of the 131 controls, only 9.9% were positive for IgM, 7.6% for IgG, 6.1% for IgA and 9.9% when an IgA and IgG combination was considered. Of 122 cases of active pulmonary tuberculosis, 41% were positive for IgM, 86.8% for IgA, 88.5% for IgG and a very high positivity (98.3%) was seen when IgA and IgG estimations were combined. A relatively low seropositivity was observed in 25 cases of pleural tuberculosis. The corresponding figures in 130 cases of extrapulmonary tuberculosis were 22.3%, 68.4%, 73.8% and 86.15%. When 60 cases of pulmonary tuberculosis who had been successfully treated with antituberculosis drugs were analyzed the rates of seropositivity fell to 11.6%, 46.6%, 58.3% and 66.6% respectively. CONCLUSION: Our findings point to a very good sensitivity (91.6%) and specificity (90.0%) of the test when combined IgA and IgG antibody titres are considered, to detect cases of adult tuberculosis. The role of IgM estimation can be restricted to the detection of cases of reactivation of tuberculosis. PMID- 7496004 TI - Rapid detection of rifampicin resistance in sputum and biopsy specimens from tuberculosis patients by PCR and line probe assay. AB - SETTING: Multidrug resistant Mycobacterium tuberculosis strains are threatening TB control in the world. Rapid diagnosis of resistance is essential for adequate treatment and optimal control of the disease. OBJECTIVE: Evaluation of a new technique (Line Probe Assay, LiPA) for easy and rapid detection of Rifampicin resistance (RMPR) of M. tuberculosis. DESIGN: After amplification of the region of the RNA polymerase, involved in RMPR, the amplified product is hybridized with a set of 10 oligonucleotides immobilized onto a membrane strip. From the pattern obtained the presence or absence of RMPR M. tuberculosis can be assessed. 67 clinical samples positive in culture for M. tuberculosis were analyzed with LiPA and results were compared with classical susceptibility testing. RESULTS: In vitro drug sensitivity testing identified 46 rifampicin sensitive and 21 resistant strains. In 65 of the 67 specimens LiPA results matched classical testing. In two RMPR cases LiPA showed a sensitive pattern. CONCLUSION: In contrast to culture and sensitivity testing, where results take on average 6 weeks, LiPA testing is an easy and rapid (< 48 h) method of detecting RMPR M. tuberculosis in clinical samples. Results correlated in 97% of the samples. In the two RMPR samples with a sensitive LiPA pattern another mechanism of resistance is suspected. PMID- 7496006 TI - Radiologic manifestations of pulmonary tuberculosis in HIV-1 and HIV-2-infected patients in Abidjan, Cote d'Ivoire. AB - OBJECTIVES: To compare the radiologic manifestations of pulmonary tuberculosis in HIV-1-infected, HIV-2-infected, and HIV-negative patients; and to assess the impact of HIV-related immunosuppression on the radiologic manifestations of pulmonary tuberculosis. METHODS: We compared chest radiographs from consecutive HIV-1-positive, HIV-2-positive and seronegative patients with pulmonary tuberculosis. Differentiation between HIV-1 and HIV-2 antibodies was based on a synthetic peptide-based enzyme immunoassay. A subset of patients had CD4+ lymphocyte levels estimated by flow cytometry; in these patients, abnormalities on chest radiographs were analysed in relation to the severity of CD4+ lymphocyte depletion. RESULTS: HIV-1-infected patients were significantly more likely to have extrapulmonary tuberculosis than were HIV-2-infected or HIV-negative patients (20% vs 8% and 9%). Among patients with pulmonary tuberculosis, no differences were observed in the rates of specific abnormalities on chest radiographs between HIV-1- and HIV-2-infected patients; both HIV-1- and HIV-2 infected patients had a higher frequency of pleural effusion than did HIV negative patients (8% and 9% vs 4%). Among HIV-infected patients with CD4+ counts of > or = 400/mm3, 200-399/mm3, and < 200/mm3, respectively, the proportions with non-cavitary infiltrates and hilar adenopathy increased significantly (33% to 44% to 58%, and 0% to 14% to 20%), while the proportion with cavitary lesions decreased significantly (63% to 44% to 29%). CONCLUSIONS: The radiologic manifestations of pulmonary tuberculosis in HIV-infected patients varied significantly over the spectrum of immune deficiency. HIV-infected patients with tuberculosis and relatively high CD4+ counts showed only slight differences from HIV-negative persons. HIV-1-positive patients had a higher frequency of extrapulmonary tuberculosis at presentation than those infected with HIV-2. Radiographic abnormalities were broadly similar in HIV-2-infected and HIV-1 infected patients. Clinicians and radiologists must be alert to the altered radiologic spectrum of pulmonary tuberculosis in immunosuppressed HIV-infected patients. PMID- 7496005 TI - Survey of knowledge, attitudes and practices for tuberculosis among general practitioners. AB - SETTING: Representative sample survey of knowledge, attitudes and practices (KAP) for tuberculosis among private general practitioners (GPs) in 1993 in Korea, OBJECTIVE: To investigate the KAP of general practitioners on the prevention and treatment of tuberculosis. DESIGN: Questionnaire surveys were performed for 923 private general practitioners through 29 health centres. RESULTS: 49% of GPs considered that the Korean tuberculosis situation is not serious. 54% were worried about infection from patients. 47% answered that BCG vaccination causes untoward reactions with no or limited effectiveness. 47% considered the National Tuberculosis Programme (NTP) unfavourably. Over 50% did not consider sputum examination essential in case finding/diagnosis, and 75% in monitoring of treatment response. For initial treatment of active tuberculosis, only 11% were prescribing the current Korean NTP's six-month standard regimen. 73% were giving currently non-recommendable regimens, and 16% unacceptably bad regimens. However, this situation could be improved, as 80% of GPs expressed the wish to acquire knowledge. CONCLUSION: Many misunderstandings were found in the field of transmission, BCG vaccination and the performance of the NTP; sputum examinations were considerably neglected in case finding/diagnosis and treatment monitoring. As for treatment, 89% were giving either non-recommendable regimens or bad regimens. PMID- 7496007 TI - Indicators and research methods for rapid assessment of a tuberculosis control programme: case study of a rural area in South Africa. AB - SETTING: An evaluation of the Tuberculosis Control Programme (TBCP) of KaNgwane, a rural, previously designated 'homeland' of South Africa, was performed in 1990. OBJECTIVE: The evaluation is presented as a case study to illustrate a proposed methodology for rapid, comprehensive, management oriented TBCP research. DESIGN: The components of the TBCP were categorised into direct TBCP activities, management processes and integration of the TBCP into Primary Health Care (PHC). The methods used to evaluate indicators for each of these components included: policy analysis, interviews with staff and patients, observation of in-patient and ambulatory patient care, record reviews and a household survey. RESULTS: The regional scope of the study yielded a wealth of useful information at a managerial level. Detailed, quantitative information was obtained, allowing for the identification of weaknesses in the TBCP and the targeting of appropriate action. Particularly problematic aspects were: the health information collection, case-finding and case-holding, health education and coordination and supervision. Comparative analyses between different health wards helped identify particular areas of weakness. Both illustrative results and methodological limitations are presented. The methodology used is related to previous South African TB health service research and to the emerging field of rapid assessment methods. CONCLUSIONS: A rapid but comprehensive review of a TBCP can be done if indicators and research methods are carefully selected and record systems are functional. If record systems are not functional, the non-record review methods still offer substantial information on many indicators and one or two quantitative indicators (e.g. case holding) could be selectively measured through reviewing a sample of records. PMID- 7496008 TI - A computer-based ordering system for supplies in national tuberculosis programs. AB - SETTING: National tuberculosis programs in need of budgeting and planning for supplies. OBJECTIVE: To assist national managers to rationally and rapidly determine needs in supplies and their costs for the national tuberculosis program. METHODS: A spreadsheet has been developed to calculate needs in supplies for tuberculosis control based on the number of nationally notified cases. The spreadsheet has 3 interdependent basic components that need to be modified. The first component requires an update of exchange rates of currencies from countries which are major suppliers for program materials. The second component contains a list of commonly used treatment regimens in national programs and their cost. The number of reported tuberculosis cases for the last complete reporting half-year on then corresponding regimens needs to be provided. The third component lists supplies for diagnosis and treatment and the costs of these to the International Union Against Tuberculosis and Lung Disease (IUATLD). The information required here includes (after updating the country-specific list of suppliers and the cost of their material) the current stock in supplies and editing the resulting suggestion for orders according to needs and convenience of unit size. RESULTS: The spreadsheet suggests the ordering size of supplies for consumption and reserve. Based on input provided by the user, it will turn out estimates of the cost of the order for supplies in diagnosis and treatment. CONCLUSIONS: Computers are coming increasingly into use at the national level of many tuberculosis programs in low income countries. This spreadsheet, available from the IUATLD, is designed to provide national managers with the background on how to calculate needs for supplies and to provide them with a simple tool to assess these needs and their costs rapidly. PMID- 7496009 TI - Control of drug-resistant tuberculosis. AB - Drug-resistant tuberculosis is a global problem, but the necessity and quality of intervention should be assessed by country since the causes of drug resistance, and thus the appropriate intervention strategies, may vary considerably between countries. An analysis of the development and spread of drug-resistant tuberculosis is presented and the respective roles of import of the disease and of HIV are discussed. A definition of inadequate treatment and a systematic approach to the quantitative and qualitative aspects of the drug resistance problem is proposed. The 'treatment delivery process' (TDP) is introduced as a diagnostic tool to analyse the causes of secondary (or acquired) resistance in a specific area. Intervention strategies should be based on the evaluation of this TDP. Intervention strategies and general aspects of their implementation are discussed. PMID- 7496010 TI - A female patient with tuberculous polyserositis, and congenital tuberculosis in her new-born child. AB - A 24-year-old woman was admitted to our department with sepsis, polyserositis and suspected abdominal tumor immediately after delivery. Effusion analyses suggested an etiology of tuberculosis. The severe general condition of the patient allowed for no invasive diagnostic procedures. Antituberculosis therapy was administrated too late and was of no benefit. The fatal outcome was due to septic shock and adult respiratory distress syndrome (ARDS). Autopsy findings confirmed tuberculous polyserositis as well as tuberculous infection of the lungs. The same diagnosis was established in her newborn infant, who responded well to antituberculosis treatment. PMID- 7496011 TI - Failure of drug penetration and acquisition of drug resistance in chronic tuberculous empyema. AB - We describe a patient with drug-resistant chronic tuberculous empyema in whom substantial differences between achievable pleural fluid and serum drug concentrations were displayed. The ratio of maximum concentration in pleural fluid to serum was especially low for rifampin (4%) but was also low for streptomycin (34%) and ofloxacin (48%). Subtherapeutic drug concentrations in the pleural fluid may have contributed to acquisition of drug resistance in this case. PMID- 7496012 TI - Congenital miliary tuberculosis with intestinal perforations. AB - The case of a male preterm baby who had congenital miliary tuberculosis with multiple intestinal perforations is reported. The patient was successfully treated with antituberculosis chemotherapy and surgical repair along with other supportive therapy. This is one of the rarest cases of the type described here. PMID- 7496013 TI - Acute dystonic reaction after Heaf test. PMID- 7496014 TI - Comparison of Heaf guns. PMID- 7496015 TI - Sensitivity of SL-IV and PPD for sero-diagnosis of tuberculosis in HIV seronegative and HIV-infected persons. PMID- 7496016 TI - The "futility debate" and the management of Gordian knots. PMID- 7496017 TI - Progress in the futility debate. PMID- 7496018 TI - Is refusal of futile treatment unjustified paternalism? PMID- 7496019 TI - The problem of medically futile treatment: falling back on a preventive ethics approach. PMID- 7496020 TI - Decedents' reported preferences for physician-assisted death: a survey of informants listed on death certificates in Utah. PMID- 7496021 TI - Empirical studies on euthanasia and assisted suicide. PMID- 7496022 TI - The limitation of empirical research in ethics. PMID- 7496023 TI - Circular questioning by ethics committees: who's asking the doctors? PMID- 7496024 TI - Of circles and lines, metaphors and narratives: toward a systems theory of ethics consultation. PMID- 7496025 TI - The ghost walks again: unpacking the assumptions of circular questioning. PMID- 7496026 TI - Incidental findings: patients' knowledge, rights, and preferences. PMID- 7496027 TI - Genetic counseling, non-directiveness, and clients' values: is what clients say, what they mean? PMID- 7496028 TI - Genetic counseling: making room for beneficence. PMID- 7496029 TI - Substituted judgment: in search of a foolproof method; a response to Baergen. PMID- 7496030 TI - Transforming or vampiric? When care-providers share their subjective realities with their patients. PMID- 7496031 TI - Mouse half embryos: viability and allocation of cells in the blastocyst. AB - We have investigated the developmental capacity of mouse embryos in which one blastomere was destroyed by lysis at the 2-cell stage. The allocation of cells to the trophectoderm and inner cell mass (ICM) was documented by differential cell counts on single embryos after 2 days under different culture conditions. Viability and further developmental potential were tested by embryo transfer to foster mothers. The conditions used were: (1) in vitro culture in modified BMOC-2 medium, (2) in vivo oviduct transfer to immature (prepuberal) females, and (3) in vivo oviduct transfer to pseudopregnant females. Half embryos almost always fared less well for all parameters of development than control embryos developing under the same conditions. Lower total cell numbers in half embryos were accounted for by decreases in both ICM and trophectoderm with a disproportionate decrease in ICM in smaller embryos. In both half and control embryos, the growth conditions affected the rate of morphological development, the total cell number, and embryo viability. Unlike the effect of halving embryos, the growth condition effects on total cell number can be accounted for primarily by differences in ICM cell number, with trophectoderm cell number remaining constant. These results provide new information on the ability of the mouse embryo to differentially regulate ICM and trophectoderm cell number under different conditions, and confirm our previous work showing the advantage of short-term development in vivo over short term in vitro culture (Papaioannou and Ebert [1986] J. Reprod. Fertil. 76:603 608).(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7496032 TI - Modulation of cell migration and vessel formation by vascular endothelial growth factor and basic fibroblast growth factor in cultured embryonic heart. AB - Vascular endothelial growth factor (VEGF) and basic fibroblast growth factor (bFGF) stimulate endothelial cell proliferation, migration, and vascular tube formation. We tested the hypotheses that these growth factors stimulate (1) cell migration and (2) assembly into cord-like structures in embryonic rat heart explants cultured on collagen gels. Atrial and ventricular explants from rat embryos at 12 (E12, avascular) and 14 (E14, early vascularization stage) days of gestation were cultured on a collagen substrate. Western blot analysis of the explants indicated that endogenous VEGF was present in both atria and ventricles during incubation. Addition of bFGF to E12 explants markedly increased cell migration, whereas VEGF had no significant effect. In E14 explants neither growth factor influenced cell migration. Cotreatment with VEGF and bFGF did not have a synergistic effect on the migration distance of cells from either E12 or E14 embryonic hearts. However, VEGF stimulated the appearance of cord-like structures in E14, but not E12, explants. Transmission electron microscopy analysis showed that these cord-like structures consist of elongated cells, some of which aggregate into clusters, or form tube-like structures, similar to capillaries. Serial sections of monolayers revealed that tube formation occurs beneath the surface of collagen gel. We conclude that in this model system VEGF and bFGF play distinct roles, at specific time points, in coronary vascular tube formation in the developing heart. PMID- 7496033 TI - Characterization of alpha 1, beta 1, and gamma 1 laminin subunits during rabbit fetal lung development. AB - Laminin-1 is an extracellular matrix protein composed of three polypeptide chains that are designated alpha 1, beta 1, and gamma 1. We investigated the expression of laminin alpha 1, beta 1, and gamma 1 subunit chains during several stages of rabbit fetal lung development. Utilizing polyclonal antibodies directed against human placental laminin and immunoblot analysis, we found that the highest levels of laminin alpha 1, beta 1, and gamma 1 subunit chains in the fetal lung were present on day 26 of gestation (term = 31 days), coincident with the initiation of alveolar epithelial cell differentiation. Levels of the laminin chains were approximately five times higher in fetal lung at day 26 of gestation than in adult lung tissue. Different temporal patterns of laminin alpha 1, beta 1, and gamma 1 subunit chain expression were observed, data suggestive that the chains are independently regulated during lung development. Laminin was localized to the basement membranes of bronchi, bronchioles, prealveolar ducts, and blood vessels in fetal lung tissue, as shown by immunostaining with polyclonal laminin antibodies. A similar staining pattern was observed in adult lung tissue, but the alveolar wall was also stained. Laminin was also observed surrounding a few mesenchymal cells in fetal lung on day 19 of gestation; the number of positive mesenchymal cells increased with lung development. Laminin alpha 1 subunit chains, detected using a monoclonal antibody, were present in the basement membranes of bronchi, bronchioles, prealveolar ducts, and blood vessels in fetal lung tissue. No laminin alpha 1 chain staining was observed in the mesenchyme of early fetal lung tissue. Using a monoclonal antibody, laminin beta 1 subunit chains were immunolocalized in the basement membranes of bronchi, bronchioles, in prealveolar ducts, and surrounding some mesenchymal cells in fetal lung tissue. Laminin alpha 1 and beta 1 subunit chains in adult lung tissue were present in basement membranes of airways, blood vessels, and alveoli. Thus, changes in the localization and accumulation of laminin near the time of alveolar type I and type II epithelial cell differentiation suggest that laminin may play a role in mediating the differentiation of these cell types during rabbit fetal lung development. PMID- 7496034 TI - Homozygosity for the Min allele of Apc results in disruption of mouse development prior to gastrulation. AB - Mutation of the APC (adenomatous polyposis coli) gene is an early event in colon tumor development in humans. Mice carrying Min (multiple intestinal neoplasia), a mutant allele of Apc, develop intestinal and mammary tumors as adults. To study the role of the Apc gene in development, we have investigated the phenotype of embryos homozygous for ApcMin (Min). Development of the primitive ectoderm fails prior to gastrulation in homozygous Min embryos. By midgestation, the presumed homozygotes consist of a mass of trophoblast giant cells with an additional cluster of much smaller embryonic cells. These results indicate that functional Apc is required for normal growth of inner cell mass derivatives. PMID- 7496035 TI - Number of adrenergic and islet-1 immunoreactive cells is increased in avian trunk neural crest cultures in the presence of human recombinant osteogenic protein-1. AB - OP-1, also known as BMP-7, is a member of the TGF-beta superfamily of proteins and was originally identified on the basis of its ability to induce new bone formation in vivo. OP-1 mRNA is found in the developing kidney and adrenal gland as well as in some brain regions (Ozkaynak et al. [1991] Biochem. Biophys. Res. Commun. 179:116-123). We have tested the effect of recombinant human OP-1 on quail trunk neural crest cultures. The number of catecholamine-positive cells which developed after 7 days in vitro in the presence of OP-1 was increased in a dose-dependent manner, with a greater than 100-fold maximal stimulation observed. The increase in the number of catecholamine-positive cells in the presence of OP 1 was paralleled by an increase in the number of tyrosine hydroxylase (TH) positive cells. In contrast, total and melanocyte cell number were unaffected by the presence of OP-1. The number of Islet-1-immunoreactive cells was also increased by OP-1, but to only about half the value seen for TH. Double label experiments revealed these Islet-1-positive cells were a subset of the TH positive cells. Inhibitors of DNA synthesis prevented the OP-1-mediated increase in adrenergic cell number, indicating that OP-1 does not act on a postmitotic cell population. However, labeling studies with bromodeoxyuridine indicated that OP-1 did not increase the proportion of the cell population engaged in DNA synthesis. Thus, the OP-1-mediated increase in adrenergic cell number most likely occurs as a result of the enhanced survival of a subpopulation of adrenergic precursors or an increase in their probability of adrenergic differentiation, but not by increasing the mitotic rate of adrenergic precursors or adrenergic cells themselves. In contrast to OP-1, TGF-beta 1 decreased adrenergic cell number. When OP-1 and TGF-beta 1 were added simultaneously, TGF-beta 1 antagonized the OP 1-mediated increase in adrenergic cell number in a dose-dependent manner. PMID- 7496036 TI - Transient expression of glutamate decarboxylase and gamma-amino butyric acid in embryonic lens fibers of the rat. AB - We have determined the localization and developmental expression of glutamate decarboxylase (GAD67) in the rat lens. Immunofluorescence experiments showed that GAD67 was transiently expressed in the nuclear fiber cells of the lens between embryonic days (E) 15 and 20, with maximal immunostaining occurring on E17 and E18. gamma-amino butyric acid (GABA) co-localized with GAD67 in the embryonic nuclear fiber cells. Reverse transcription-polymerase chain reaction (RT-PCR) tests showed that at least three alternatively spliced forms of GAD67 mRNA, including mRNAs with and without the I80 and the I86 insert, were transiently co expressed with GAD67 in the embryonic lens. The major GAD67 protein in the lens was 67 kDa. We conclude that enzymatically active GAD67 is transiently expressed in the lens nuclear fiber cells of the embryonic rat. The transient expression is regulated by transcriptional and/or posttranscriptional processes. We speculate on the basis of possible common gene regulatory elements for glutamate and ornithine decarboxylases and the involvement of these enzymes with polyamine synthesis, that the transient expression of GAD67 may be connected to nuclear and/or DNA breakdown during lens fiber cell differentiation. PMID- 7496037 TI - Uncoupling histogenesis from morphogenesis in the vertebrate embryo by collapse of the transneural tube potential. AB - We have shown that unidirectional pumping of Na+ out of the neural tube's luminal fluids in amphibian embryos produces a large potential difference (40-90 mV, lumen negative to the abluminal surface). This transneural tube potential (TNTP) is analogous to the Na+ dependent transepithelial potential (TEP) that exists across surface ectoderm. This TEP is retained in ectoderm after it is internalized when the neural folds fuse to form the neural tube. The TNTP can be markedly reduced for several hours by injection of the Na+ channel blockers amiloride or benzamil into the lumen by iontophoresis through microelectrodes. Here we describe the effect of TNTP modification on developmental anatomy. Axolotl embryos possessing a fused and closed neural tube (stage 21-23) were injected with either amiloride or benzamil and allowed to continue development for 36-52 hr. These were compared to control embryos injected with vehicle alone, or to embryos in which amiloride or benzamil was iontophoresed just beneath surface ectoderm. All embryos in which the TNTP was reduced were grossly defective. These were characterized by a disaggregation of the cells comprising the structures that had already begun to form (otic primordia, brain, spinal cord, notochord) as well as a failure in the development of new structures. Remarkably, some of these embryos displayed continuing development of external form in the complete absence of concomitant internal histogenesis. We discuss the ways in which a large endogenous voltage gradient associated with an epithelial potential difference (the TNTP) may be required both for the structural integrity of the early neuroepithelium, and a prerequisite for normal morphogenesis. PMID- 7496038 TI - Chondrocyte apoptosis in endochondral ossification of chick sterna. AB - In the process of endochondral ossification, chondrocytes progress through a series of maturational changes, including division and hypertrophy, that culminate in chondrocyte loss and cartilage resorption. From an investigation of morphology, DNA fragmentation and collagen synthesis in the developing chick sterna we have characterized chondrocytes death in this process. Light microscopy of resorbing sterna demonstrated chondrocyte condensation at the interface with the invading vasculature and electron microscopy demonstrated a range of chondrocyte morphologies, including retraction from the pericellular matrix, cytoplasmic and nuclear condensation, and vesiculation suggestive of sequential changes characteristic of apoptosis. Isolation and end-labeling of DNA from chick primary ossification centers demonstrated fragmentation to nucleosome sized units, only in primary ossification centers exhibiting active resorption, and in situ detection of DNA fragmentation showed a restriction to chondrocytes at the interface with invading blood. We conclude that terminal differentiation of chondrocytes results in death by an apoptotic process prior to resorption of the tissue and invasion by blood vessels. The extent of DNA fragmentation correlated closely with the proportion of cells displaying a condensed phenotype in contralateral primary ossification centers and peaked at an early stage of resorption, suggesting that chondrocyte apoptosis may be an initiating event in tissue resorption and vascular invasion. Comparison of DNA fragmentation with expression of the hypertrophic chondrocyte phenotype, as indicated by type X collagen synthesis, suggested that DNA fragmentation was a late event in the process of chondrocyte hypertrophy and probably corresponded with chondrocyte condensation PMID- 7496039 TI - Thrombospondin-4 is expressed by early osteogenic tissues in the chick embryo. AB - The thrombospondins are a family of related glycoproteins found in the embryonic extracellular matrix. To date, five members of this family have been identified. Thrombospondin-1 and thrombospondin-2 have similar primary structure, but are expressed in different tissues at different times during development. Thrombospondins-3, -4, and cartilage oligomeric protein belong to a second thrombospondin subgroup in which the carboxyl-half of each molecule is most similar to thrombospondin-1 and -2. Here we report the cloning and sequencing of a novel probe to avian thrombospondin-4. We have used this probe to determine the origins of thrombospondin-4 in the chick embryo by in situ hybridization. Thrombospondin -4 transcripts first appear in the mesenchyme surrounding bone anlage coinciding with the initial stages of osteogenesis. The expression in osteogenic tissues is transient: thrombospondin-4 mRNAs are not seen in the osteoblasts of bone collars in developing long bones. This pattern is distinct from avian thrombospondin-2 which is expressed in perichondrium and embryonic fibrous connective tissues. Our observations indicate that connective tissues are the principal site of thrombospondin-4 expression in the chick. The diverse origins of different thrombospondin gene family members imply distinctive roles for these proteins related to the growth and differentiation of cartilage, tendons, and bone. PMID- 7496040 TI - Embryonic expression of tenascin-X suggests a role in limb, muscle, and heart development. AB - Tenascin-X (TN-X) is the newest member of the tenascin family of extracellular matrix proteins and it is highly expressed in muscular tissues during development. To gain insight into the possible functions of TN-X during development, we evaluated its expression in the rat embryo. Using an 800 bp cDNA encoding the fibrinogen-like domain of TN-X, we show that TN-X expression begins in migrating cells of the epicardium in the E12 heart. The epicardium provides progenitors of fibrous and vascular tissue to the developing heart. After the epicardium is complete, TN-X is expressed in the sub-epicardial space in association with developing blood vessels, and later by non-myocytes dispersed through the myocardial wall. A similar pattern of TN-X expression, first in connective tissue surrounding muscle, and then by a subset of cells within muscle, was seen in para-axial, body wall, craniofacial, and appendicular muscle. This pattern suggests a role in connective tissue cell migration and late muscle morphogenesis. TN-X is also highly expressed in the interdigital space at E15 and surrounding developing tendons, suggesting an additional role in cell fate determination. Although the pattern of TN-X expression is distinct from that of tenascin C, they are frequently expressed in close proximity. Indirect genetic evidence in humans suggests an essential function for TN-X, and the pattern of TN X expression in heart, skeletal muscle, and limb is consistent with this hypothesis. PMID- 7496041 TI - Depolarization-induced 86Rb+ efflux in CHO cells expressing a recombinant potassium channel. AB - Cells expressing a recombinant human voltage-activated potassium channel (K channel), Kv1.5, have been used in a functional assay that measures depolarization-stimulated 86Rb+ efflux as an indicator of K-channel function. Neither untransfected nor vector-transfected cells display measurable 86Rb+ efflux under depolarizing conditions. The depolarization-induced 86Rb+ efflux is blocked by standard K-channel blockers quinine, 4-aminopyridine and 3,4 diaminopyridine, but not by tetraethylammonium, quinidine, glibenclamide, or several peptide toxins. The pharmacological profile of the recombinant system reflects that reported for the channel in its native state. In such a system with no observable endogenous background, analysis of recombinant K-channel subtypes allows rapid assessment of pharmacological agents with isoform selectivity and specificity. Inclusion of compounds of unknown activity in an assay such as this could identify agents capable of modulating specific K-channel isoforms. Development of this high through-put assay system for the study of specific isoforms is a critical step in the identification and development of drugs that affect the desired target tissues with predictable pharmacology and minimal side effects due to nonselective K-channel interaction. PMID- 7496042 TI - Influence of hematocrit and platelet count on impedance and reactivity of whole blood for electrical aggregometry. AB - Previous studies have shown that differing qualities of blood specimen seem to influence whole blood electrical aggregometry (WBEA), making it difficult to standardize the method. The aim of this study was to investigate the impact of hematocrit (HCT) and platelet count (PLC) on in vitro platelet aggregation in citrated whole blood (CWB) in order to compensate for their possible effects on impedance aggregometry. Red blood cells and blood platelets were isolated from fresh citrated whole blood taken from 15 healthy donors (mean age = 26 years) and recombined to 20 physiologically relevant combinations of hematocrit and platelet count (HCT: 20-50, PLC: 100-500). Platelet aggregability was measured using WBEA with three different triggers. A special-purpose software package was used in this study, ensuring proper calibration, acquisition, and evaluation of analogue to digital converted data, allowing the calculation of a set of characteristic parameters of each impedance curve. Most of the linear regressions showed that all parameters significantly depend on HCT and PLC. Furthermore, we found interactions of both variables, making it impossible to focus on the effects of one of the investigated variables only. The outcome of this study is a set of dependences, allowing the calculation of regressions for in vitro aggregation in whole blood, enabling a comparison of blood of any quality with each other, regardless of the variables HCT and PLC. Together with the previously defined dependence of sample age on WBEA data, this step should help to make this technique a more reliable and practicable clinical tool, making it suitable for daily routine investigations. PMID- 7496043 TI - Hot plate versus tail flick: evaluation of acute tolerance to continuous morphine infusion in the rat model. AB - The development of tolerance to continuous morphine infusion (2, 4 and 6 mg x kg( 1) x hr(-1) was assessed in rats using two different methods for evaluation of nociception, tail flick (TF) and hot plate (HP). The influence of repeated testing on nociception was evaluated using two regimens; series 1 was tested repeatedly 1, 2, 4, 6, and 8 hr after initiating the morphine infusion and series 2 was tested only twice, at maximum morphine effect and at 8 hr. Both, TF and HP showed pain threshold elevation after the morphine administration of 4 or 6 mg x kg(-1) x hr(-1), which reached a maximum at 2 hr after the start of the infusion. HP: reduction of the effect was found in group 4 mg x kg(-1) x hr(-1) in the series subjected to repeated testing; group 6 mg x kg(-1) x hr(-1) showed reduced effect in both sides. TF: the response latencies did not show reduction at 8 hr. Since TF is predominantly a spinal response and HP is predominantly supraspinal, the results suggest that tolerance during the first 8 hr of morphine infusion develops mainly at supraspinal level. PMID- 7496044 TI - Radioimmunoassays for cyclic AMP cross-react with phosphodiesterase inhibitors and buffer components. AB - We addressed the issue of cross-reactivity of several commonly used phosphodiesterase inhibitors with radioimmunoassays for cyclic AMP, after we had observed a considerably high cross-reactivity with a noncommercial antibody. Theophylline, pentoxifylline, penthydroxifylline (BL 194), albifylline (HWA 138), torbafylline (HWA 448), A 80 2715, isobutyl methylxanthine, and the nonmethylxanthines amrinone and rolipram were dissolved in supplemented and boiled cell culture medium (RPMI 1640). These samples were assayed for apparent cyclic AMP in two different, commercially available radioimmunoassay kits (based on polyclonal antibodies), applying the nonacetylated protocol. Cross-reactivity was dose-dependent and nonlinear. Samples containing theophylline and amrinone exhibited the strongest cross-reactivity in assay A (NEN/DuPont): 3.0 +/- 0.5( nM) and 2.4 +/- 1.1 (-nM) apparent cyclic AMP +/- SD at 1-nM spike, respectively. With the more sensitive assay B (Amersham), higher concentrations of apparent cyclic AMP were detected: from 7.9 +/- 0.4 nM (for albifylline) to 3.5 +/- 0.1 nM (for rolipram). Values were calculated from standard curves set up in the respective assay buffer, where culture medium controls resulted in 1.8 +/- 0.3 nM and 3.1 +/- 0.1 nM for assay A and B, respectively. The culture medium interference increased with rising cyclic AMP concentrations. Although comparatively low, this degree of cross-reactivity is relevant for in vitro experiments. Phosphodiesterase inhibitors are commonly administered at millimolar concentrations, and resulting cyclic AMP levels are often in the nanomolar range. Neglecting these findings may lead to falsely high readouts of cyclic AMP concentrations. PMID- 7496045 TI - Use of resampling techniques to estimate the variance of parameters in pharmacological assays when experimental protocols preclude independent replication: an example using Schild regressions. AB - Estimates of variance in pharmacological assays are usually made by repeating the experiment with different tissues. Biological factors, such as the inability to wash a drug from tissue, may preclude the type of replication that is appropriate for the statistics of interest. For example, in Schild regressions, replication is usually done at each concentration of antagonist. In some test systems, replication of dose-response curves is not possible. For example, some persistent agonists cannot be removed from tissues after exposure, while in other systems, rapid desensitization severely alters tissue sensitivity to repeated challenge with agonist. In this paper, we demonstrate how a statistical resampling method, bootstrapping, can be used to derive estimates of the confidence intervals for pA2, pKB, and slope from Schild plots. This method utilizes the speed of the computer to estimate variance by repeatedly resampling the data. The advantage to this method is that it can be used for many different experimental designs. For a data set obtained from a Schild regression of atenolol antagonism of isoproterenol in the guinea pig left atrium, bootstrap estimates of confidence limits were calculated for cases where dose ratios were derived from the same tissue and randomly paired tissues. These estimates showed good agreement with estimates obtained using conventional analytical methods, thus suggesting that this method may be useful in practice. PMID- 7496046 TI - Isolation of syncytial microvillous membrane vesicles from human term placenta and their application in drug-nutrient interaction studies. AB - The initial step in placental uptake of nutrients occurs across the syncytial microvillous membrane of the trophoblast. This study was designed to isolate syncytial microvillous membrane vesicles (SMMV) of human term placenta, to validate their purity and viability, and to investigate the interaction of several commonly used drugs with the transport of two essential nutrients: alanine and choline. SMMV were isolated according to an established procedure, but instead of homogenization the initial preparation step was replaced by mincing of placental tissue followed by gently stirring to loosen the microvilli. These modifications doubled the protein recovery and increased the enrichment in alkaline phosphatase, whereas no substantial contamination with basal membranes nor interfering subcellular organelles was found. The functional viability of the vesicles was evaluated through the transport of alanine. In accordance with literature, uptake was sodium-dependent, inhibitable by structural analogues, and saturable. A number of cationic drugs were were able to able to inhibit choline uptake, whereas no effect on alanine transport was observed. Anionic drugs, drugs of abuse, and catecholamines did not interfere with alanine transport either. In conclusion, our isolated SMMV provide a suitable tool for screening drug-nutrient interactions at the level of membrane transport. In view of the very low susceptibility of the alanine transporter to drug inhibition and the relatively high drug concentrations necessary to inhibit choline transport, it seems unlikely that clinically important drug interactions may occur with these nutrients. PMID- 7496049 TI - Hypertension. PMID- 7496048 TI - Assessment of intestinal permeability changes induced by nonsteroidal anti inflammatory drugs in the rat. AB - Intestinal permeability was investigated as an alternative to intestinal ulceration for measuring nonsteroidal anti-inflammatory drug (NSAID) gut damage in the rat and developed as a method for routine measurement. NSAID dose-response curves produced using the two indices of damage showed that intestinal permeability is as sensitive and reproducible as ulceration, although changes could not be detected before visible ulceration occurred. Lactulose, [51Cr]-EDTA and [14C]-carboxyinulin were compared as possible in vivo markers of rat intestinal permeability. Measurement of [51Cr]-EDTA permeation was found to be the most sensitive and reproducible method. Dose-response curves produced by measuring [51Cr]-EDTA permeation were used to compare the potency of the two NSAIDs piroxicam and (S+) ibuprofen; piroxicam was found to be 10 times more potent in increasing intestinal permeability than (S+)-ibuprofen. These studies show that intestinal permeability measurement is a useful alternative to other methods of assessing NSAID adverse effect and is easily and rapidly performed. PMID- 7496047 TI - An easy and economical method to prepare cells for cytologic analyses. AB - A crucial step in any cytology application begins with the placement of cells on slides in a manner that provides good preservation of cell morphology. Commercial cytocentrifuges are the most commonly used devices for this purpose. However, there are instances when these devices are either not available or not applicable. We devised an easy and economical alternative method to prepare cells for cytologic analysis. The device (cytograv) was easily assembled from common laboratory and office supplies, and produced cell preparations of similar quality as those produced with a cytocentrifuge (cytospin-3). Cellular analyses of biological fluids is one of the most common applications for these kinds of devices. The cytograv device was successfully employed in the identification and verification of a PMN isolation procedure from whole blood. The cytograv device was also successfully used to quantitative increases in the number of PMNs in bronchoalveolar lavage fluid recovered from mice treated with increasing doses of paraquat. These two examples illustrate some of the many possible uses of the cytograv device to provide high-quality preparations for cytologic analyses. PMID- 7496050 TI - Diseases of the aorta, pulmonary, and peripheral vessels. PMID- 7496051 TI - Newer antihypertensive drugs. AB - We review three new classes of antihypertensive drugs: imidazolines, monatepil, and neutral endopeptidase inhibitors. Imidazolines are a new generation of central acting drugs. Because of their greater affinity for imidazoline receptors rather than alpha 2-adrenoreceptors, they cause less fatigue and dry mouth than the older central acting drugs. Monatepil is a single drug with both calcium antagonist and peripheral alpha 1-antagonist properties. This agent not only lowers blood pressure but also lowers serum cholesterol, an effect mediated by its alpha-antagonism, and has anti-lipid peroxidation properties. Monatepil might become an effective antiatherosclerosis drug. Neutral endopeptidase inhibitors lower blood pressure by inhibiting the metabolism of atrial natriuretic peptide and other vasoactive peptides. PMID- 7496052 TI - Hypertensive heart disease and heart failure. AB - Hypertension is associated with the remodeling of left ventricular geometry and abnormalities of function that may precede geometric changes. Rather than a specific disease, "hypertensive heart failure" is a spectrum of disorders that result from left ventricular geometric changes and comorbid conditions. Heart failure may present as abnormalities of diastolic or systolic function, although symptoms (dyspnea, fatigue) and physical findings (edema, rales) may be similar. PMID- 7496053 TI - Hypertensive heart disease and the diabetic patient. AB - Patients with diabetes mellitus are particularly vulnerable to cardiovascular disease. Although structural and functional myocardial complications are present in patients with diabetes alone, they are particularly severe in patients with both diabetes and hypertension. Considerable evidence--both in experimental animal models and in humans--points to hypertension as of critical importance in the pathogenesis of severe diabetic heart disease. In diabetic hypertensive cardiomyopathy, coronary artery disease as well as structural and functional abnormalities are more pronounced than would be expected from either process alone. The myocardial damage is attributed mainly to hypertension, whereas the myocellular dysfunction is attributed mainly to diabetes. Together, the consequences to the myocardium are devastating. Strict control of the hypertension and diabetes may have an ameliorative effect on the subsequent development of diabetic heart disease. PMID- 7496054 TI - Reversal of hypertrophy: an active biologic process. AB - Evidence from in vivo, in vitro, and genetic studies suggests that the reversal as well as the development of left ventricular hypertrophy do not depend solely on hemodynamic load; other factors are involved. Several humoral agents that may affect mitogenesis of cardiac myocytes and nonmyocitic elements have been identified, including the local renin-angiotensin system, norepinephrine, endothelins, transforming growth factor beta, insulin-like growth factor, bradykinin, prostaglandins, and nitric oxide. Animal studies using various models of left ventricular hypertrophy are beginning to suggest that reversal of hypertrophy may decrease mortality, improve coronary flow reserve, and maintain cardiac performance. Studies in humans are less supportive, and more are needed before it may be concluded that reduction of left ventricular mass decreases the cardiovascular morbidity and mortality associated with cardiac hypertrophy. PMID- 7496055 TI - The interface of hypertension and ischemic heart disease. AB - Myocardial infarction remains the single most common cause of death in patients with essential hypertension. This becomes particularly evident when the hypertension is associated with left ventricular hypertrophy. To combat the continuing high mortality from myocardial infarction in hypertensive heart disease, however, all aspects of the relationship must be studied. Thus, addressing the interface from an epidemiological standpoint as well as from a pathological point is critical and progress in these areas as well as in areas of management are ultimately likely to lead to a fall in morbidity and mortality from ischemic heart disease in patients with hypertension. PMID- 7496056 TI - New diagnostic inroads in hypertension and atherosclerosis. AB - In the past two to three decades, the practice among hypertension specialists has been to lower blood pressure with the strong assumption that morbidity and mortality would be reduced. This was largely based on large clinical trials done in the 1960s and 1970s that did not include angiotensin-converting enzyme inhibitors or calcium-channel blockers. Performing large clinical trials is expensive and the results often do not apply to individual patients. Until recently, noninvasive methods to monitor end-organ effects of elevated blood pressure and other cardiovascular risk factors were limited. The development of high-resolution ultrasound and evolving magnetic resonance technology will allow early detection of heart and blood vessel changes and will make monitoring of the progression and regression of disease possible. This will allow better selection of treatment programs for each patient. PMID- 7496057 TI - Endothelin in hypertension. AB - The endothelins, 21-amino-acid peptides produced by the endothelium of blood vessels and many other tissues such as the kidney, brain, endocrine organs, etc., are potent vasoconstrictors, and are also endowed with mitogenic and cell hypertrophic properties. Endothelin may be involved in the pathogenesis of hypertension through vascular, renal, endocrine, and neural effects. Although many studies have been performed to test the hypothesis that these peptides have a pathophysiologic role in hypertension, it is only recently that evidence has been found of enhanced production of endothelin-1 in some models of hypertension, particularly in blood vessels in deoxycorticosterone acetate-salt hypertensive rats. Vascular responses to endothelin-1 have been shown to be normal or depressed in many models of experimental hypertension, and also in humans with essential hypertension. Elevation of blood pressure and development of vascular hypertrophy is blunted in deoxycorticosterone acetate-salt hypertensive rats treated chronically with endothelin receptor antagonists. Spontaneously hypertensive rats do not overexpress vascular endothelin, and do not exhibit a hypotensive response to chronic endothelin receptor antagonism. Malignant spontaneously hypertensive rats treated with deoxycorticosterone acetate and salt exhibit vascular overexpression of endothelin-1 and respond to endothelin antagonists with lowering of blood pressure. A genetic role of components of the endothelin system has been suggested in Dahl salt-sensitive rats. In human essential hypertension, there is as yet little evidence of activation of the endothelin system. A role of endothelins in hypertension is thus becoming increasingly apparent in severe forms of experimental hypertension, but further studies are required to establish whether these peptides are involved in the human disease. PMID- 7496058 TI - The natriuretic peptides in hypertension. AB - The natriuretic peptide family consists of three members: atrial natriuretic peptide, brain natriuretic peptide, and C-type natriuretic peptide. Atrial and brain natriuretic peptides possess similar effects, causing natriuresis, vasodilation, and suppression of the renin-angiotensin-aldosterone system. C-type natriuretic peptide has been suggested to exert its predominant effect on the vasculature, eliciting vasodilation and inhibiting the proliferation of vascular smooth muscle cells. Numerous studies have broadened our current knowledge of the regulation of natriuretic peptide gene expression, biosynthesis, and secretion, as well as structure of specific receptors. This has led to a better understanding of the renal, cardiovascular, and endocrine actions of natriuretic peptides in both normal and pathophysiological states, including hypertensive disease. Development of nonpeptide neutral endopeptidase inhibitors and antagonists for natriuretic peptide receptors may reveal the range of potential therapeutic application of atrial and other natriuretic peptides in hypertension. PMID- 7496059 TI - Management of aortic intramural hematoma. PMID- 7496060 TI - Management of aortic dissection. AB - Patients with aortic dissection are divided into two main groups: those with hypertension and those with syndromes involving hereditary defects of the aortic wall. The common finding in the setting of both Marfan syndrome and hypertension is dilatation of the aorta. The biological events that may lead to dissection are a decrease in the holding power of the internal layer, an increase in blood pressure, or an increase in aortic diameter combined with a decrease in wall thickness. Treatment of established acute type A dissection is still associated with a hospital mortality of greater than 10% and long-term complications from persistent blood flow in the distal false lumen. The main therapeutic challenge is to prevent aortic dissection by screening of aortic root diameter in patients at risk, together with life-long beta-blockade for Marfan patients. In patients with suspected dissection, the preference is for rapid noninvasive diagnosis and early surgical repair to reduce the risk of malperfusion and irreversible organ damage. Results of surgery have improved, with an increasingly conservative approach to repair of the aortic valve with biological glues. Repair is now routinely carried into the aortic arch using improved methods of cerebral protection such as retrograde cerebral perfusion through the superior vena cava. Nevertheless, there remains scope for further innovation in the prevention and treatment of this difficult problem. PMID- 7496061 TI - Update on carotid endarterectomy. AB - Progress in therapeutic decision-making for patients with carotid artery disease has come from evaluations made by several major randomized clinical trials. For patients who are symptomatic and have arteriographically proven stenoses of 70% or more, endarterectomy is clearly established as the treatment of choice. Symptomatic patients with stenoses of less than 70% remain the subject of ongoing study. Asymptomatic patients are at low risk of stroke save for those with the highest (80% to 90%) degrees of stenosis, and even for these subjects the annual stroke rate is close to the operative risk. Four trials have been reported but the practicing physician is still without clear guidelines. The appropriateness of endarterectomy remains unsettled for any group of patients with narrowing of the carotid artery in the absence of symptoms. The decision to apply endarterectomy to symptomatic and to asymptomatic patients must attend to not only the prognostic importance of the degree of stenosis, but also to the vascular risk profile of the individual patient. PMID- 7496062 TI - Severe pulmonary hypertension in congenital heart disease: a review of Eisenmenger syndrome. AB - Patients with Eisenmenger syndrome are characterized by severe irreversible pulmonary hypertension and reversed central shunt. In the past, congestive heart failure was generally considered common and prognosis poor. However, newer information suggests that congestive heart failure is uncommon and that the natural survival of patients with Eisenmenger syndrome is better than survival achieved with transplantation. In this review, I discuss who gets Eisenmenger syndrome, when they get it, why they get it, and what the consequences are for adults with the disorder. PMID- 7496063 TI - Recent advances in the diagnosis and treatment of coarctation of the aorta. AB - Although it has been the topic of intense medical and surgical attention for over 50 years, coarctation of the aorta continues to be a major cause of cardiovascular morbidity and mortality in infants and children. Refinements in established diagnostic modalities--primarily fetal, transesophageal, and intravascular echocardiography--have improved pre- and post-treatment assessments. Aggressive and early intervention, whether by surgery, catheter, or both, have been shown to increase initial success; the results of long-term follow-up are yet to be determined. The best possible outcome requires prompt recognition and effective treatment directed at immediate gradient relief, which will hopefully prevent or reduce the known late and long-term sequelae. PMID- 7496064 TI - Preoperative assessment of the patient with cardiac disease. AB - Assessment of the patient with cardiovascular disease is a challenging problem, especially for the subset of patients undergoing surgery. Perioperative cardiac morbidity affects more than 1.5 million people annually in the United States, and consumes more than +20 billion per year in health care resources. Solution of the problem initially had been addressed using routine historical and clinical evaluations and risk indices. More recently, nonroutine preoperative cardiovascular testing has been recommended, including exercise and Holter electrocardiography, radionuclide ventriculography, stress echocardiography, and dipyridamole thallium scintigraphy. Each of these tests has strengths and limitations that are addressed in this review. Paradigms for preoperative assessment of high-risk patients, based on the presence of coronary disease and the functional status of the patient, are presented and discussed. Diagnostic and therapeutic approaches for patients at high risk remain challenging; the mandate is for large-scale clinical trials to evaluate both the therapeutic and cost effectiveness of new preoperative assessment strategies. PMID- 7496065 TI - Chronic venous insufficiency. AB - The consequences of chronic deep venous insufficiency are a major medical concern and result in significant loss of human productivity in addition to a significant compromise of lifestyles. The postphlebitic syndrome of chronic venous stasis and ulceration is a result of chronic venous hypertension and reflects a sequela of events occurring secondary to venous valvular insufficiency and reflux. Diagnostic techniques have advanced considerably; however, the prevention and management of this entity continue to be inadequate. Many new areas are being explored, both from a medical and a surgical perspective, and it is hoped that as clinical investigation proceeds, new conceptual approaches and techniques will arise to combat this difficult medical problem. The literature reviewed in this article reflects a spectrum of research attempting to understand the basic underlying hemodynamic as well as cellular and tissue changes that contribute to the development of postphlebitic signs and symptoms. More investigation is needed to enable us to proceed from the descriptive understanding of this entity to the mechanisms that result in this disease state. Preventive and curative management needs to succeed the current palliative approach to therapy. PMID- 7496066 TI - Magnesium deficiency and parathyroid function. PMID- 7496067 TI - Rapid detection of Mycobacterium tuberculosis in sputa by the amplification of IS6110. AB - A polymerase chain reaction (PCR) assay for the rapid detection of Mycobacterium tuberculosis in sputum samples was studied. The target DNA was a 123-base pair (bp) fragment of IS6110, which was repeated in the M.tuberculosis genome and was specific for the M.tuberculosis complex. Glass beads (2mm diameter) and lysozyme were used to lyse the mycobacteria and DNA was extracted by the phenol-extraction method. The amplified PCR product was detected by examination of ethidium-bromide stained agarose gel and by hybridization with an oligonucleotide alkali phosphatase-labeled probe. A total of 70 samples were tested. PCR was positive in all 13 smear and culture-positive samples, in 5 of 8 smear-negative and culture positive samples, and in 1 of 49 smear and culture negative samples. The overall sensitivity and specificity were 85.7% and 98%, respectively. Thus, IS6110 as a PCR target was found to be very useful for the rapid diagnosis of M.tuberculosis infection and start of anti-tuberculous chemotherapy. PMID- 7496068 TI - Progression of coronary artery disease in patients receiving percutaneous transluminal coronary angioplasty: angiographic comparison of angioplasty and non angioplasty sites. AB - Long-term follow-up of coronary artery stenosis was carried out in 81 patients undergoing successful percutaneous transluminal coronary angioplasty (PTCA). Progression of coronary stenosis was observed in 38 patients with 52 sites, and regression was seen in 23 patients with 23 sites during 2 years or more of follow up. Progressive change occurred in three sites of 97 PTCA lesions and 49 of 1,090 non-PTCA lesions (no significant difference). On the other hand, regressive change was observed in 19 sites of PTCA lesions and four of non-PTCA lesions (p < 0.05). The percentage of narrowing in PTCA lesions was 28.0 +/- 14.4% at 3-6 months after PTCA, and this improved to 22.8 +/- 15.8% at late study angiography. Coronary score changes were -0.16 +/- 0.54 in PTCA lesions, and 0.08 +/- 0.46 in non-PTCA lesions (p < 0.01). We concluded that the progression of obstructive coronary artery disease in sites with previous PTCA is less than in those with non-PTCA. PMID- 7496069 TI - Neurohumoral factor responses to mental (arithmetic) stress and dynamic exercise in normal subjects. AB - We examined the responses of plasma catecholamine and plasma renin activity (PRA) during mental (arithmetic) stress and dynamic exercise in 15 normal subjects. Compared to rest values, there was a significant increase in the plasma epinephrine concentration (E), but not in the plasma norepinephrine concentration (NE) or PRA during mental arithmetic. During dynamic exercise, there were significant increases in both NE and E and in PRA. The ratio of E to NE was significantly higher during mental arithmetic than during dynamic exercise. NE was significantly correlated with PRA at rest, during mental arithmetic and dynamic exercise. No significant correlations were observed between E and PRA. Dynamic exercise primarily induces a sympathetic nervous response, and mental (arithmetic) stress mainly induces an adrenal response. The renin-angiotensin aldosterone system response may in part be regulated by sympathetic nervous activity during both mental (arithmetic) stress and dynamic exercise. PMID- 7496070 TI - Retrospective analysis of the multidisciplinary treatment for 534 hepatocellular carcinoma patients over a 12-year period. AB - Multidisciplinary treatments for 534 patients with hepatocellular carcinoma (HCC) over a 12-year period from 1981 to 1992 were retrospectively studied. Cumulative survival curve of all patients showed a fine hyperbolic curve, and 1-year, 3-year , 5-year survivals were 66, 33, and 18%, respectively. Survival of female patients was better than male patients, and survival of HBsAg-negative patients was better than that of HBsAg-positive patients. Several background factors such as the Child-Pugh classification, liver function tests or factors, number of tumors, and the size of the main tumor were analyzed, and each factor was related to survival. Survival curves of 3 treatments, hepatectomy, percutaneous ethanol injection (PEI), and transcatheter arterial embolization (TAE)+PEI, overlapped to a large degree. Also the survival curve of the more recent 6-year period from 1987 to 1992 was better than that of the earlier 6-year period from 1981 to 1986. PMID- 7496071 TI - Effects of bathing in hot water on portal hemodynamics in healthy subjects and in patients with compensated liver cirrhosis. AB - Bathing in hot water induces vasodilatation of the peripheral vessels and alteration of blood distribution. Using a Doppler flowmeter, we evaluated the effect of bathing in hot water on portal hemodynamics. Eight controls and 8 patients with compensated liver cirrhosis remained immersed in hot water at 42 degrees C for 5 minutes. Portal flow significantly decreased from 811 +/- 141 (ml/min) at the baseline to 530 +/- 98 in the controls and from 855 +/- 308 to 642 +/- 208 in cirrhotics immediately after bathing. This decrease (25%) in cirrhotics was significantly less than that (35%) in controls because vessel diameter did not decrease as rapidly as it did in controls. Portal flow after 30 minutes returned to the baseline values in controls but remained low in cirrhotics. These findings suggest that cirrhotics have a lower hemodynamic response to bathing in hot water compared with that in controls. PMID- 7496072 TI - A family with hereditary serum cholinesterase deficiency. AB - A family with serum cholinesterase (SChE) deficiency is reported. A 64-year-old woman was admitted for the excision of colon adenoma; her laboratory data revealed a markedly decreased level of SChE. SChE genes of the patient and her family members were amplified by the polymerase chain reaction (PCR) and analyzed by direct sequencing. The patient's SChE gene had a homozygous frame shift mutation, in which an extra adenine was inserted in codon 315 (ACC-->AACC), resulting in the appearance of a new stop codon in codon 322. The family study disclosed that her brother and sister had the same frame shift mutations in homozygote and heterozygote, respectively. PMID- 7496073 TI - Synchronous fluctuation of interleukin-6 and platelet count in cyclic thrombocytopenia and thrombocytosis. AB - We describe a case of cyclic thrombocytopenia and thrombocytosis, whose cytokine levels, granulocyte-macrophage colony stimulating factor (GM-CSF) and interleukin 6 (IL-6) in plasma, fluctuated in synchrony with platelet count. The levels of the two cytokines correlated significantly with the platelet count for 11 observations over an 8-month period (r = 0.79, p < 0.01 for GM-CSF and r = 0.87, p < 0.001 for IL-6). No inverse relationship between platelet-associated immunoglobulin G (PAIgG) and platelet count was observed (r = 0.39, p > 0.20). These findings suggest that the fluctuation of platelet count in this case may result from an aberration of the cytokine network regulating megakaryopoiesis and platelet formation. PMID- 7496074 TI - An adult diagnosed as hyper-IgM immunodeficiency syndrome. AB - A 27-year-old male who visited our hospital because of pneumonia was diagnosed as hyper-IgM immunodeficiency syndrome. His serum IgM level was markedly elevated, while the serum level of IgD was normal with a markedly decreased level of serum IgG and IgA. The proportion of T and B cells of peripheral blood lymphocytes was normal. However, B cells bearing surface IgG or IgA were not detectable by immunofluorescence technique. There was a consanguineous marriage in his family, suggesting that his disorder was caused by a genetic abnormality such as X-linked recessive and also autosomal recessive inheritance, although further study is necessary. CD40 ligand cDNA did not appear to contain any abnormal changes within the coding region. PMID- 7496076 TI - IgG and IgM rheumatoid factor levels parallel interleukin-6 during the vasculitic phase in a patient with Churg-Strauss syndrome. AB - A 43-year-old woman with a 10-year history of bronchial asthma developed marked peripheral blood eosinophilia and symptoms of vasculitis. A diagnosis of Churg Strauss syndrome (CSS) was made, and her symptoms were successfully treated with low-dose prednisolone. Serum rheumatoid factors (RF) of both IgG and IgM, but not IgA or IgE, subclasses transiently appeared in accordance with the vasculitic phase. Serum interleukin-6 (IL-6) levels also transiently increased in this phase and the peak level was reached just prior to the maximum of RF elevation, suggesting the role of IL-6 as an inducer of RF. RF and IL-6 seemed to be involved in the pathogenesis of the vasculitis in this patient. PMID- 7496077 TI - Multiple endocrine neoplasia type 1 accompanied by duodenal carcinoid tumors and hypergastrinemia: a familial case. AB - A 38-year-old female was admitted for investigation of the cause of hypergastrinemia, hypercalcemia and an elevated plasma parathyroid hormone (PTH) level. Her siblings, elder brother and sister who had duodenal carcinoid tumor with hypergastrinemia and parathyroid adenomas, were diagnosed as multiple endocrine neoplasia (MEN) type 1. In the present patient, endoscopic examination showed a carcinoid tumor of the duodenum, but examinations by computed tomography (CT) and ultrasonography of the abdomen failed to reveal pancreatic lesions. Serum gastrin levels of this patient and her siblings improved to the normal level after resection of carcinoid tumors. The hypergastrinemia accompanying MEN type 1 in these cases might be due to carcinoid tumor of the duodenum. PMID- 7496075 TI - Streptococcal toxic shock syndrome: report of two cases. AB - Two Japanese cases of streptococcal toxic shock syndrome (STSS) are reported. The first patient was a 45-year-old male who developed necrotizing fasciitis and myositis of the left thigh, refractory hypotension, hepatic dysfunction and acute renal failure; the patient died despite treatment. Streptococcus pyogenes was isolated from the inflamed fascia. The second patient was a 69-year-old female who had coagulopathy, polymyositis and hepatic function abnormality. Streptococcus pyogenes was isolated from blood culture. She was immediately placed on high-dose ampicillin as well as other supportive measures, and she survived. PMID- 7496078 TI - Successful prophylaxis of wheat-dependent exercise-induced anaphylaxis with terfenadine. AB - A 20-year-old female was brought to our emergency unit with generalized erythema and discomfort induced by running after having eaten wheat bread. The laboratory examinations, including eosinophils, total IgE, RAST score to wheat, heat challenge test and methacholine inhalation test were within normal limits. No anaphylactoid responses occurred after provocation tests of wheat bread intake or exercise alone. However, on provocation exercise test after eating pancakes, she developed hypotension, generalized itching and urticaria associated with an elevation of plasma histamine levels. These findings suggested wheat-dependent exercise-induced anaphylaxis. This was completely prevented by daily administration of terfenadine 120 mg p.o. without side effects such as fatigue or drowsiness. PMID- 7496079 TI - Granular cell tumor in the ascending colon. AB - We describe a case of multiple polypoid lesions that occurred in the colon of a 62-year-old man. Eight of the lesions were demonstrated by barium enema, and five were examined histologically. A small nodule in the submucosa of the ascending colon was histologically identical to granular cell tumor, characterized by nested growth of large tumor cells with ample granular cytoplasm and small round nuclei. Immunohistochemically, the cells were positive for S-100, neuron-specific enolase and vimentin, and negative for alpha-smooth muscle actin and desmin. PMID- 7496080 TI - Somatostatinoma of the pancreas associated with von Hippel-Lindau disease. AB - A 39-year-old man was admitted because of lumbago, vomiting and massive gastrointestinal bleeding. Oliguria developed a few days later, which was followed by hyperkalemia and cardiac arrest. Autopsy disclosed multiple renal cell carcinomas with diffuse metastasis to the liver, adrenal gland, psoas muscle and vertebrae. In addition, a somatostatinoma was found in the pancreas. From these findings and past history of cerebellar hemangioblastoma and spinal hemangioma he was diagnosed to have von Hippel-Lindau disease. Von Hippel-Lindau disease with islet cell tumor is very rare and is reported here with a review of literature. PMID- 7496081 TI - Renal refractoriness to phosphaturic action of parathyroid hormone in a patient with hypomagnesemia. AB - A 50-year-old tetanic woman with hypomagnesemia is described. She had partial resection of the stomach and the jejunum at the age of 20 years. Lack of parathyroid hormone (PTH) function was indicated by hypocalcemia, hyperphosphatemia and high tubular reabsorption of phosphate. However, both plasma concentration of PTH and nephrogenous cAMP were normal. Administration of magnesium sulfate completely normalized serum phosphate and tubular transport of phosphate with only a modest increase in nephrogenous cAMP. The present findings suggest that phosphaturic action of PTH is impaired in magnesium deficiency and that steps distal to cAMP production may be responsible for the renal refractoriness to the hormonal action. PMID- 7496083 TI - Thiamine deficiency and pulmonary hypertension in Crow-Fukase syndrome. AB - A 57-year-old woman with Crow-Fukase syndrome presented thiamine deficiency and pulmonary hypertension of unknown etiology. After oral administration of prednisolone and thiamine, echocardiogram showed marked improvement of the pulmonary hypertension. To our knowledge, this is the first case of this syndrome associated with thiamine deficiency and precapillary pulmonary hypertension, which may play a role in the pathogenesis of polyneuropathy and heart failure of this syndrome. PMID- 7496082 TI - Mitochondrial encephalomyopathy with A to G transition of mitochondrial transfer RNA(Leu(UUR)) 3,243 presenting hypertrophic cardiomyopathy. AB - In a 24-year-old woman with mitochondrial encephalomyopathy presenting hypertrophic cardiomyopathy, microscopical examination of myocardial biopsy specimen disclosed severe vacuolar degeneration of myocardium and aggregates of enlarged mitochondria with proliferated cristae. Limb muscle biopsy specimen showed "ragged-red fibers" light microscopically and enlarged abnormal mitochondria with markedly increased cristae ultrastructurally. Mitochondrial DNA analysis by polymerase chain reaction (PCR) revealed an A-to-G transition in the mitochondrial transfer RNA(Leu)(UUR) gene at nucleotide position 3,243 which is reported to be associated with mitochondrial myopathy, encephalopathy, lactic acidosis, and stroke-like episodes (MELAS). However, the clinical features of this case, presenting mainly cardiac abnormalities, were not consistent with the typical MELAS. PMID- 7496084 TI - Budgerigar breeders' hypersensitivity pneumonitis presenting as chronic bronchitis with purulent sputum. AB - A 49-year-old woman complained of cough and a purulent sputum (about 40 ml/day). She kept budgerigars for a period of 30 years, she first noticed a productive cough 10 years previously. Her chest roentgenogram revealed a diffuse reticulonodular pattern. She was diagnosed as having a chronic type of budgerigar breeder's lung associated with chronic bronchitis. Peptostreptococcus anaerobius was cultured from her sputum, bronchoalveolar lavage fluid, as well as from droppings of her budgerigar. This is the second case of hypersensitivity pneumonitis presenting as chronic bronchitis with much sputum in Japan. PMID- 7496085 TI - Type II citrullinemia associated with neutropenia. AB - A 37-year-old Japanese man was admitted with delirium and hyperammonemia. He was diagnosed as having type II citrullinemia because of an elevated citrulline level on amino acid analysis and very low hepatic argininosuccinate synthetase activity. He also showed a low neutrophil count and a low serum level of granulocyte colony-stimulating factor. Reduced production of this cytokine and/or impairment of its feedback regulation by the neutrophil count may have played a role in the neutropenia of this patient. PMID- 7496086 TI - Two cases of pheochromocytoma diagnosed histopathologically as mixed neuroendocrine-neural tumor. AB - We treated two rare cases of pheochromocytoma which were histopathologically diagnosed as mixed neuroendocrine-neural tumor (MNNT): a 35-year-old male patient associated with ganglioneuroblastoma and cutaneous neurofibromatosis and a 42 year-old male patient with ganglioneuroma. Both patients showed typical clinical manifestations of pheochromocytoma without any familial traits. Although each of the diseases has its own entity and clinical features, these tumors are all derived from the neural crest tissues. The tumorigenesis of MNNT is still unknown. Here, a brief review of the recent literature on this subject is discussed. PMID- 7496087 TI - Isolated adrenocorticotropic hormone deficiency associated with growth hormone deficiency and empty sella. AB - A 38-year-old man had an acute onset of consciousness loss, pyrexia and hyponatremia. Plasma Adrenocorticotropic Hormone (ACTH) and cortisol levels were low. Plasma ACTH failed to respond to corticotropin-releasing hormone (CRH) and insulin-induced hypoglycemia whereas i.m. injection of ACTH-Z raised plasma cortisol. Plasma insulin-like growth factor-I (IGF-I) and urine growth hormone (GH) concentrations were also low and plasma thyroid-stimulating hormone (TSH) level was rather elevated. Plasma IGF-I and TSH levels were not completely normalized by glucocorticoid (GC) replacement alone although plasma GH responses to pharmacological stimuli were normalized. The GC replacement in combination with daily s.c. injection of recombinant human GH (rhGH) not only normalized plasma IGF-I and IGFBP-3 levels, but also further lowered the plasma TSH level, possibly due to an increased T4/T3 conversion, which resulted in a beneficial change in body composition. PMID- 7496088 TI - Successful hematopoietic reconstitution with granulocyte colony-stimulating factor in a patient with hypoplastic acute myelogenous leukemia. AB - We describe a 68-year-old Japanese male with hypoplastic acute myelogenous leukemia (AML) who achieved complete hematological reconstitution following granulocyte colony-stimulating factor (G-CSF) administration. The patient had pancytopenia and the bone marrow was hypocellular with 19 to 36% peroxidase positive blasts without morphological abnormalities suggestive of myelodysplasia. After receiving G-CSF as a supportive therapy for pneumonia, the blood count became normal and the bone marrow was normocellular with less than 5% of blasts. Without subsequent chemotherapy, he relapsed as a form of overt leukemia and died of pneumonia. Chemotherapy may be necessary to maintain remission in hypoplastic AML after hematopoietic reconstitution by G-CSF. PMID- 7496089 TI - Undifferentiated carcinoma in the cardioesophageal junction which produces parathyroid hormone related protein. AB - A 68-year-old man with undifferentiated carcinoma occurring in the cardioesophageal junction accompanied by hypercalcemia is reported. The serum level of parathyroid hormone-related protein (PTHrP) was remarkably elevated. Serum calcium and PTHrP levels decreased following chemotherapy, but this amelioration was temporary. He died of hypercalcemia. On autopsy, a significant amount of immunoreactive PTHrP was detected in the tumor tissue extract, and the tumor cells were stained strongly positive for PTHrP by immunohistochemistry. This is the first case of undifferentiated carcinoma in the gastrointestinal tract which demonstrated hypercalcemia due to PTHrP produced by the malignant tumor. PMID- 7496090 TI - Bendectin: review of the medical literature of a comprehensively studied human nonteratogen and the most prevalent tortogen-litigen. AB - OBJECTIVE: to review the extensive literature pertaining to the reproductive and teratogenic effects of Bendectin and the opinions of the scientific experts for the defense and plaintiff. These data were evaluated with regard to the reproductive risks of Bendectin providing a scientific framework for evaluating the views of the experts in the Bendectin litigation. DESIGN: the Bendectin literature was primarily obtained from articles cited in Research Alert of the Institute for Science Information. Other articles were obtained from Medline, review articles, and colleagues. An attempt was made to be all-inclusive, citing and reviewing all articles related to each subject being discussed. The literature includes epidemiologic studies, animal studies, in vitro studies, and basic science articles related to the principles of teratology and reproductive toxicology. Review articles, meta analyses, editorials, commentaries, articles in the press, and case reports were also included. METHODOLOGY: the methodology utilized for the evaluation of Bendectin teratogenicity was presented. It consists of a five-part analysis of epidemiologic studies, secular trend analysis, animal studies, dose-response relationships, and biologic plausibility. CONCLUSION: the five-part analysis of Bendectin reproductive effects indicates that therapeutic use of Bendectin has no measurable teratogenic effects. Presentations by many of the plantiff's experts failed to meet the scientific standards that should be expected of knowledgeable scientists and contributed to the persistence of Bendectin litigation. PMID- 7496091 TI - The use of an in vitro sperm activation assay to detect chemically induced damage of human sperm nuclei. AB - A human sperm activation assay (HSAA) has recently been developed to assess sperm quality and functionality. The HSAA involves incubating lysolecithin permeabilized sperm in Xenopus laevis ova cytoplasmic extract. Factors in the extract activate sperm nuclei; the activation involves sperm chromatin decondensation, DNA synthesis, and chromatin recondensation. We report that human sperm chemically damaged in vitro by treatment with a reversible crosslinker, ethylene glycolbis(sulfosuccinimidylsuccinate; SEGS), display abnormal chromatin decondensation when analyzed in the HSAA. Less than 20% of SEGS-treated sperm fully decondensed, vs. 97% of control sperm. Chemical reversal of the crosslinks by treatment with 5 microM hydroxylamine restored full decondensation in 76% of treated sperm. These results demonstrate that chemically damaged sperm respond abnormally in the HSAA, and that chemical damage to sperm nuclei can be detected using the HSAA. Thus, there is potential for the HSAA to be used to detect chemical alterations of sperm nuclei from men exposed to environmental toxicants. PMID- 7496092 TI - Testicular germ cell toxicity caused by vinylcyclohexene diepoxide in mice. AB - Vinylcyclohexene diepoxide (VCD) produces ovarian toxicity in female mice and rats, whereas testicular damage occurs only in mice. The objectives of these studies were to determine the target cell(s) and spermatogonial survival following VCD administration. In addition, the effects of 4-vinylcyclohexene (VCH) and two epoxide metabolites, vinylcyclohexene 1,2-monoepoxide and VCD were compared. Male mice were dosed daily with VCD (320 mg/kg/d, i.p.) and killed at 5, 10, 15, 20, 25, or 30 d. Two groups were dosed daily for 30 d and allowed to recover for 30 or 60 d. Decreases in testis weight began at 5 d and continued to 30 d. These decreases corresponded to progressive necrosis of germ cells. After 5 d of VCD, there was loss of Type I and B spermatogonia in Stages II to VI and of preleptotene spermatocytes in Stages VI to VIII. After 30 d of dosing, seminiferous tubules were devoid of germ cells except for spermatogonial stem cells. Following 30 d of recovery, 100% of the seminiferous tubules were repopulated. Epididymal spermatozoa were present after 60 d of recovery. Increasing doses of VCD (0 to 320 mg/kg/d) resulted in increasing testicular toxicity. Neither VCH (800 mg/kg, i.p.) nor VCM (200 mg/kg, i.p.) caused testicular damage. VCD administration initially results in destruction of spermatogonia and spermatocytes, which are undergoing DNA synthesis and cell replication, followed by loss of maturing cells. Neither VCH nor VCM caused testicular germ cell destruction, although all three compounds destroy germ cells in female mice. Therefore, further investigation will be necessary to understand these differences in chemical-induced toxicity between ovaries and testes. PMID- 7496093 TI - All-trans-retinoic acid-induced variant patterns of palatal rugae in Crj:SD rat fetuses and their potential as indicators for teratogenicity. AB - We determined the critical period for cleft palate formation in the presence of all-trans-retinoic acid (RA) and defined anomalous patterns within the variants of palatal rugae. The total incidence of variant patterns of palatal rugae in rat fetuses treated with RA at 40 mg/kg on gestation days 13 and 14 dramatically increased with concurrent induction of cleft palate. Correspondence analysis connected the variant patterns of palatal rugae, such as median discontinuity, extra peaks, supernumerary ruga (extra type), shortness, fusion, and bifurcation, with cleft palate. Therefore, these variants were concluded to be anomalous. The incidence of the anomalous patterns of palatal rugae was about 10 times that of cleft palate, and consequently, these anomalies might be sensitive indicators of the teratogenicity of a chemical. PMID- 7496094 TI - Neonatal exposures to technical methoxychlor alters ovaries in adult mice. AB - Technical grade methoxychlor (MXC) is an estrogenic pesticide currently used for pest control in the US. To determine the long-term effects of technical MXC on ovaries and estrous cycles, neonatal mice received 14 daily intraperitoneal injections of sesame oil only, 10.0 micrograms estradiol-17 beta, or 0.05 mg, 0.1 mg, 0.5 mg, or 1.0 mg of technical MXC. At 3, 6, and 12 months, vaginal smears were examined for 12 d and ovaries collected. All technical MXC doses and estradiol increased the duration of vaginal cornification but only estradiol, 0.5, or 1.0 mg technical MXC induced ovarian atrophy, relative ovarian weight loss, and depletion of corpora lutea. Technical MXC doses of 0.05 or 0.1 mg produced the opposite effects; the ovaries remained heavy, large, and filled with corpora lutea. On the other hand, follicular cysts were recorded in all groups, except the 1.0 mg MXC group. These augmentary and inhibitory effects of MXC at low and high doses mimic the estrogen effects at low and high doses, and were probably due to the altered hypothalamic-hypophyseal function. PMID- 7496095 TI - The reproductive and developmental toxicity profile of beta-cyclodextrin in rodents. AB - A 3-generation study with two mating phases per generation and a teratology phase was performed in the rat to assess the reproductive and developmental toxicity of beta-cyclodextrin (beta-CD) in the diet. Transient neonatal growth retardation occurred with 5% beta-CD: a similar, but equivocal, effect was also observed with 2.5%. No permanent defects or other indications of developmental toxicity were found. There was no significant maternal toxicity. The dietary level of 1.25% was found to be a NOAEL for developmental toxicity. Further investigations showed the growth retardation to be specific to dietary administration during lactation: it was not produced by parenteral (i.p.) administration and was not influenced by treatment of the dams or litters during gestation. Slight maternal nutritional deficiency, caused by physico-chemical interactions of beta-CD with nutrients in the gut is proposed as the mechanism of action. No difference in milk composition was found. PMID- 7496096 TI - Ovulation induction by clomiphene and neural tube defects. PMID- 7496097 TI - Glycol ethers in pesticide products: a possible reproductive risk? PMID- 7496098 TI - [Expert assessment of unconventional treatment methods within the scope of legal health insurance--problems based on current court decisions]. AB - Court decisions of the Federal Court for Problems of Social Welfare (Bundessozialgericht) make health insurance pay for unconventional methods under certain circumstances. Physicians of the Health Insurance Medical Service (MDK) have to examine whether these conditions are fulfilled in a special case. The specific problems they encounter there are being described in this paper. Hermeneutic differences can be noticed. To guarantee the quality of medical care, which is the duty of the statutory health insurances, seems to be difficult in the case of "unconventional" medical methods. PMID- 7496099 TI - [Between individual therapy and political intervention: campaigns against "diseases of civilization" between 1920 and 1960]. AB - The concept of "disease of civilization" is commonly used for a variety of conditions. It implies a criticism of the "modern" way of life. This criticism and the strategies put forward to prevent diseases of civilization are correlated to the specific historical context in which they are formulated. The paper analyses three positions characterising different periods in 20th century German history: In the Weimar Republic, representatives of paediatrics and social hygiene asked for the expansion of public health care programmes according to the principles laid down by the Weimar constitution. During the time of National Socialism, more radical measures were propagated by exponents of hygiene and psychotherapy, aiming at a fundamental restructuration of society in accordance with the Nazi ideology. After 1945, strategies focused on the individual: Physical exercise, abstinence, and individual psychotherapy were propagated to prevent or treat the consequences of life in modern society. PMID- 7496100 TI - [Quality management in the hospital with special reference to the medical departments]. AB - Present German health legislation requires standardisation of systems and procedures not only for medical departments but for entire clinics as well. Changes in organisation and treatment possibilities should enable clinic administrations to better manage the quality of the services provided. This system of quality control and quality management is based on the introduction of ISO 9004, part 2. Medical and non-medical procedures are standardised and streamlined through flow-diagrams, the details of which are summarised in a series of quality handbooks with an emphasis on guidelines and standards. These handbooks are distributed clinic-wide, and although acceptance is not yet determined, objections are probable. Future analysis will show the effects due to quality management in the clinical setting. PMID- 7496101 TI - [The Bremen primary examination program for refugees (I)]. AB - In April 1993, the Bremen State Office of Public Health initiated a programme based on the requirements of political asylum legislation while answering the need of medical care for refugees in the first phase of their stay. Newly arriving asylum seekers channelled into legally obligatory "initial accommodation" are offered to avail themselves of a regular medical consultation in their accommodation facility. Thus, they have the opportunity to resort to medical examination, a doctor's advice, and, if need be, initial treatment. PMID- 7496104 TI - [Evaluation of wood preservative pollutants of indoor air]. AB - The evaluation of indoor pollution by wood preservatives and pesticides has been a matter of increasing interest for the past years. The present paper reviews actual knowledge. The so-called "wood preservative syndrome" is referred to. Basics and evaluation criteria concerning the investigation of external exposure (wood, dust, air) and human biomonitoring is summarized. Several substances, in particular pentachlorophenol, lindane and pyrethroids, and principle possibilities to decrease exposure are considered. PMID- 7496103 TI - [The "Freiburg Spinal Study"--epidemiology of occupationally-induced spinal diseases of health care employees in Germany]. AB - Musculoskeletal diseases are very common in modern societies. As several international investigations have shown that the etiology of these diseases is correlated with working conditions in some professions. With financial subsidy by the German insurance company for the regulation and compensation of occupational diseases in the health care sector (BGW), we investigated in 6 different projects the possible relation of working conditions and musculoskeletal diseases among health care professionals, esp. nurses. Our contribution describes the design of the different sub-projects and presents some results. The summarising discussion points out that there was a strong tendency in all sub-projects to confirm the hypothesis that nurses run a significantly higher risk to suffer from work related musculoskeletal diseases and disorders. The results of the sub-project "cohort study in nursing pupils" support this hypothesis. The results do not only supply important background information for the process of compensating occupational diseases in this field but also indicate a tremendous personal and structural need for more and better prevention. PMID- 7496105 TI - [Retrospective evaluation of western blot for detection of early HIV infections]. AB - The purpose of HIV diagnosis is to establish safely whether there is an infection or not. The enzyme-linked immunosorbent assay (ELISA) as screening test, and the western blot assay for confirmation is the most widely used serologic test system to get this information. Diagnostic problems occur if the two tests yield different results. In our 1993 study 491 (4.4%) of 11,127 tested sera were reactive by ELISA. 39 (7.9%) of these samples could not be confirmed by western blot, giving negative or indeterminate results. In addition, 370 ELISA-negative samples were tested by HIV-1 western blot to detect the infection in the early stage. 115 (31%) of these sera showed indeterminate western blot patterns, the other samples were negative. Results of follow-up investigations of 26 (Table 1) or 11 (Table 2) persons with indefinite serodiagnosis were analysed. HIV infection was not detected in any of these cases. In the literature, persons whose sera were reactive according to ELISA and were indeterminate in western blot, had a 3-5% probability of an aisting HIV infection. In contrast, indeterminate western blot patterns in ELISA-negative sere were without significance for the prognosis as to whether a person was or was not HIV infected. PMID- 7496106 TI - [New methods for managing substance abuse by the public health office]. PMID- 7496102 TI - [The Bremen primary examination program for refugees (II)]. PMID- 7496109 TI - [Common recommendation of specialty representatives for social medicine of German medical faculties]. PMID- 7496107 TI - ["Public health at universities and in public health service" memorandum: perspectives for cooperation]. PMID- 7496108 TI - [Comments on theory development in "public health". Berlin Public Health Research Committee--partial projects "Goals and goal indicators for public health in East and West Berlin"]. AB - Due to extensive government funding, public health research in Germany presently goes through times of revival and relative prosperity. The multiprofessionality of its background, its nearly disrupted scientific tradition in Germany, and the idiosyncrasies in their understanding of "health" among different professions impede the development of a common scientific identity. This paper underscores the importance of semantic precision and the avoidance of pretentious, overstrained theories in favour of pragmatic scientific approaches. Complementary to its action-oriented rationale, the German public health community should more than ever devote itself to health objectives and outcomes. PMID- 7496111 TI - [Comment on Wassermann, O., H. Kruse: Health risks of emissions from refuse incinerators]. PMID- 7496110 TI - [Comment on Wassermann, O.,H. Kruse: health risks of emissions from refuse incinerators]. PMID- 7496114 TI - [Objectivity, reproducibility and validity of 3D ultrasound in dermatology]. AB - Recently it has become possible to visualize the three-dimensional (3D) architecture of sonographic structures using high-resolution ultrasound above 20 MHz (Dermascan C, Denmark; DUB 20, Germany). We studied the objectivity, reproducibility, and validity of these two 3D analysis systems on 22 skin tumors. To validate both systems, we compared the sonometric and histometric volume in 4 tumors. The objectivity was calculated by repeated evaluation of the volume on the same serial B-scan sections at 3-month intervals. The two-dimensional B-scans and 3D images exhibited similar characteristics. Volumetric evaluations were reproducible with both systems (variation coefficient < or = 9.9%), when the applicator was oriented longitudinal to the body axis. By changing the orientation, the coefficient increased up to 58.1% (DUB 20). Sonometric and histometric volumes of a phantom model and of the tumors exhibited small deviations in the Dermascan C (< or = 8.1%) and in some tumors large deviations in the DUB 20 (< or = 93.1%). We can conclude that the 3D sonography is a reproducible method, which can be used to evaluate the volume of skin tumors and metastases under chemotherapy or radiotherapy. We suggest in particular to use the in-vivo tumor volume as a prognostic parameter for the malignant melanoma. PMID- 7496113 TI - [The value of 3D video endoscopy for surgical procedures]. AB - We discuss the problems of spatial perception that arise by the transmission of endoscopic surgery by traditional video systems and analyse the improvements that can be achieved by two- und three-dimensional systems. Starting with a description of the physics of vision and the principles of two- and three dimensional imaging, we demonstrate the importance of optimal adaptation of technology to the natural perception processes of the observer. In this context we review critically the technology that is presently available. Only stereoscopic video technology with 'shutter glasses' provides the observer with the spatial information that is of such decisive importance for minimally invasive surgery. Finally we discuss new technical developments that further improve three-dimensional vision in endoscopy, such as a 3D video module that is small enough to be used intracorporally. PMID- 7496115 TI - [Ultrasound angiography: phantom measurements of slow blood flow]. AB - Power Doppler ultrasound is a color-coded Doppler ultrasound method based on the strength of the Doppler shift signal. The sensitivity for slow flow was tested with different flow phantoms. The lowest detectable velocity was 0.04 cm/s in a water- or oil-filled tank at an angle of 45 degrees and 0.24 cm/s in an oil medium at an angle of 86 degrees. The pulse repetition frequency (PRF) was 100 kHz at the lowest detectable velocities, the acquisition time depended mostly on the attenuation of the medium. The new method depended very little on the angle of insonation. Under ideal conditions detectable flow velocities are within the range of mean capillary flow velocity. PMID- 7496112 TI - [Imaging methods in diagnosis and differential diagnosis of breast cancer]. AB - The diagnosis of breast cancer is primarily based on X-ray mammography. Under optimal conditions, a sensitivity of approximately 90% can be achieved. When strict criteria of indication are observed for the additional use of ultrasound or contrast-enhanced MRI, the sensitivity can be increased to about 98%. In addition, the differential diagnosis between benign and malignant lesions can be improved and the rate of biopsies due to false-positive mammograms can be reduced. However, further investigation with ultrasound or MRI of dense or mastopathic breasts that are clinically asymptomatic is not indicated, since it reduces specificity without significant gain of sensitivity. PMID- 7496116 TI - [Reliability of ultrasound ileus diagnosis]. AB - We investigated in this study if we would be able to find the correct indication for laparotomy earlier by sonographical examination than by clinical and radiological findings. In 48 of 594 patients with unknown abdominal diseases we found an intestinal obstruction by laparotomy. The indication based on clinical and radiological findings in 37 patients (sensitivity: 77.1%). In comparison the indication for laparotomy was predetermined correctly only after sonographical examination in 45 patients (sensitivity: 89.6%). According to the number of sonographical findings the reliability of the indication for laparotomy increased. The examination by sonography is a valuable tool in the diagnosis of intestinal obstruction. PMID- 7496117 TI - [Paranasal sinus mycetoma with orbital involvement in a patient with AIDS]. AB - A patient with AIDS was hospitalized with a left-sided face swelling and protrusion of the bulbus. After cranial computed tomography and fine-needle aspiration biopsy of the fossa temporalis we diagnosed a mycetoma; localisation and histology made an aspergilloma most probable. Antimycotic therapy led to complete remission of the symptoms. Post mortem we only could culture Candida albicans out of the abscess cavity. PMID- 7496119 TI - Age, sex and transaminase dependency of specific and nonspecific results from enzyme immunoassays for antibodies to hepatitis C virus and follow-up of blood donors. AB - BACKGROUND: Second-generation enzyme immunoassays (EIA-2) for antibodies to hepatitis C virus (anti-HCV) have an improved specificity and sensitivity compared to first-generation enzyme immunoassays (EIA-1). Therefore the question arises how many anti-HCV-positive blood donors were missed by the EIA-1, how many were false positive, how false-positive donors should be dealt with and how the results of the EIA-2 correlate with demographic data and surrogate testing for serum alanine aminotransferase (ALT). MATERIAL AND METHODS: A total of 208,544 individual North German blood donors not preselected for anti-HCV negativity were tested for anti-HCV with EIA-2 and, if repeatedly reactive (rr), with a licensed supplementary test (RIBA-2). RESULTS: Overall, 0.43% of the donors were EIA-2 rr, but only 0.12% of women and 0.09% of men were RIBA-2 positive. RIBA-2 positivity rates were very low in donors 18 to 27 years old (0.03% and 0.05%) and clearly rose with age in women but not in men. The rate of unspecifically positive EIA-2 results (entirely negative in RIBA-2) rose with age in both sexes and did not correlate with ALT. The ALT distribution was age dependent with a completely different pattern for men and women. Anti-HCV positivity was strongly correlated with ALT albeit on a very low level: more than 97% of donors with strongly elevated ALT were anti-HCV negative. Follow-up and comparison of EIA-1, EIA-2 and RIBA-2 results for the subsequent donations showed that only 8% of now RIBA-2 positive donors were not detected by EIA-1. Apparent seroconversions in EIA-2 are usually not specific: only one out of 66 apparent seroconversions could be confirmed by RIBA-2. 0.15% of the donor population showed an inconsistent EIA-2 pattern during follow-up. CONCLUSIONS: We therefore suggest that donors should not be excluded from further donations on the basis even of multiple EIA-1 positive results or on the basis of only one EIA-2-positive donation. The value of ALT screening for transfusion safety should be reconsidered. PMID- 7496118 TI - [Extraskeletal Ewing sarcoma: differential diagnosis of a soft tissue tumor]. AB - We report on a 30-year-old man from Vietnam who complained about diffuse pain in the left epigastrium. Using the ultrasound a pleural effusion in the left recessus costodiaphragmaticus was found. The chest X-ray showed a paravertebral soft-tissue tumor at the left side, with CT and NMR a sarcoma was suspected. The patient was treated by local resection of the tumor, histologically an extraosseous Ewing's sarcoma was identified. Due to the following chemotherapy and local radiation therapy no local recurrence and metastases have developed for 32 months. PMID- 7496121 TI - [Screening for increased cholesterol values in blood donors]. AB - OBJECTIVE: In a pilot study blood donors were screened on HDL, LDL, and total cholesterol. The practicability and significance of such a screening in a blood bank should be tested. DESIGN: For this purpose, current methods to determine HDL, LDL, and total cholesterol were adapted on microtiter plates and validated. In a period of about 4 weeks all blood donors were tested for the three cholesterol parameters. SETTING: Red Cross blood bank in Baden-Baden, Germany with about 150,000 donations per year. PARTICIPANTS: The HDL, LDL, and total cholesterol results of 10,006 blood donors could be evaluated. INTERVENTIONS: None. RESULTS: The mean total cholesterol was 192 +/- 36 mg/dl, the mean HDL cholesterol 56 +/- 15 mg/dl, and a mean LDL cholesterol of 123 +/- 39 mg/dl was found. The known age-dependency of the measured cholesterol fractions could be confirmed in our blood donor population. According to the current recommendations, more than 20% of the tested blood donors need a therapy. CONCLUSIONS: The study shows that the screening of blood donors for increased total cholesterol for prevention of coronary heart disease could be done with a minimum of costs. PMID- 7496120 TI - [Experiences with anti-Rhesus-D therapy in pretreated patients with idiopathic thrombocytopenia]. AB - OBJECTIVE: We tested the effect of anti Rhesus D [anti Rh(D)]-specific IgG in heavily pretreated patients with idiopathic thrombocytopenic purpura (ITP). DESIGN: Retrospective single case studies. SETTING: Clinical department of hematology. PATIENTS: 6 consecutive patients with heavily pretreated therapy refractory ITP. INTERVENTIONS: 5 patients received one cycle of Anti Rh(D) in doses between 1,200 and 6,000 micrograms in 1 patient 2 consecutive cycles were applied. Treatment effect, durability, and side effects were monitored. RESULTS: Patients after splenectomy and/or immunosuppressive therapy did not respond. Response was short-lived in 2 other patients, one long-term remission could be achieved. Responders showed slight decreases in hemoglobin indicating mild hemolysis. Other major side effects were not observed and the therapy was well tolerated. CONCLUSIONS: Our results suggest that therapy with Anti Rh(D) is safe and comparably inexpensive. No clear dose/effect correlation was found in our investigation. Only patients with platelet sequestration into the spleen might respond to Anti Rh(D) therapy. PMID- 7496122 TI - The gel test: investigation into the aetiology and the significance of a positive auto-control. AB - BACKGROUND: Shortly after the gel test was introduced into routine immunohaematology, an increased percentage of patients were reported to show a positive auto-control in the indirect antiglobulin test (IAT) of uncertain significance as the direct antiglobulin test (DAT) in the tube technique was negative. MATERIALS AND METHODS: In our study 13,280 randomized patient blood samples were screened and additional investigations carried out including an analysis of patient histories in the 97 blood samples that were auto-control positive in the gel test. RESULTS: In 87.4%, a re-testing with polyspecific antiglobulin serum (83.2% with anti-IgG) showed positive results in contrast to only 52.9% re-tested by the tube test. Neither nonspecificity nor cold agglutinins were significant. None of the patients examined showed any signs of haemolysis except for one with pernicious anaemia. We concluded that the increased number of positive auto-controls and DATs is due to the greater sensitivity of the gel test and thus the detection of minute quantities of specific cell-bound IgG molecules, i.e. warm auto-antibodies or drug-induced antibodies. CONCLUSION: Prior to transfusion, a positive result should be confirmed by a tube DAT. If this test is negative and there is no history of a previous transfusion or of haemolysis, the transfusion should not be delayed by carrying out further time-consuming investigations. PMID- 7496123 TI - Plateletpheresis-induced increase in platelet reactivity using different cell separators. AB - OBJECTIVE: Since plateletpheresis is being used increasingly, it is important to regard quality control to check health risks for donors and to exclude these. DESIGN: Controlled randomised prospective open comparative study. SETTING: Department of Transfusion Medicine of a University Clinic. PARTICIPANTS: 112 platelet donors were examined. INTERVENTIONS: Prior to and after plateletpheresis platelet reactivity was determined. The platelet concentrates in the two groups of 56 donors each were produced using either the cell separator 'CS-3000' and the collecting chamber PLT 30TM with the Omnix system (group I) or the cell separator 'AS-104' (group II). RESULTS: In group I five donors showed a pathologically increased platelet reactivity (p = 0.1297) after plateletpheresis. In group II there were 10 donors with a pathologically increased platelet reactivity (p = 0.0046) after plateletpheresis. The mean concentration of platelets was reduced by separation using the CS-3000 Omnix from 238 +/- 49 x 10(3)/microliters to 172 +/- 32 x 10(3)/microliters (68 +/- 27 x 10(3)/microliters) and from 243 +/- 53 x 10(3)/microliters to 180 +/- 31 x 10(3)/microliters (63 +/- 33 x 10(3)/microliters) using the AS-104. In the first case the platelet yield was 3.9 x 10(11) platelets/concentrate, in the latter case it was 2.9 x 10(11) platelets/concentrate. The 'CS-3000 Omnix' is significantly more effective in separating (58.4 +/- 15.5%) than the 'AS-104' with 44.2 +/- 7.2% (p < 0.0001). CONCLUSIONS: Since both donor groups were comparable regarding all factors recorded--especially the cardiovascular risk factors--the separation process could be responsible for the different traumatisation of platelets. PMID- 7496124 TI - Comparison of different protocols in plateletpheresis with the haemonetics MCS 3p blood cell separator with respect to parameters of product quality. AB - BACKGROUND: In thrombocytapheresis, intermittent flow cell separators produce platelet concentrates (PC) with a higher leucocyte contamination than continuous flow blood cell separators. The new discontinuous flow cell separator Haemonetics MCS 3p offers a low-leucocyte solution for PC. The quality of PC obtained by the MCS 3p was investigated in this study. DESIGN: Prospective study. SETTING: Haemapheresis Unit of a University Clinic. PATIENTS: Healthy blood donors from the haemapheresis unit. MATERIALS AND METHODS: Platelet (PLT) yield, separation efficiency and white blood cell (WBC) contamination were studied in three different protocols. Two protocols used a haemocalculator, which calculated the target volume based on the donor's physical characteristics and the desired PLT yield for the procedure. Protocol I used 3,000 ml as target process volume and protocol II 3.3 x 10(11) as desired PLT yield. Protocol III was used without haemocalculator. Glucose, lactate, lactate dehydrogenase (LDH), morphology score and pH value were analysed to investigate the quality of the PC. RESULTS: Platelet yield and separation efficiency were not statistically different in the three protocols. Leucocyte contamination was lowest in the protocol without haemocalculator (median: 3.15 x 10(6), range 0.4-20.8 x 10(6)). Glucose, lactate, LDH and pH were not statistically different in the three protocols. Morphology score was best in protocol III. CONCLUSIONS: PLT collection with the MCS 3p blood cell separator results in sufficient thrombocyte yields. Using the haemocalculator we were not able to achieve the desired platelet yield. For this reason, and because of the higher WBC contamination in protocol II we prefer PLT collection without the haemocalculator. The quality of the platelet concentrates was good with respect to the parameters glucose, lactate, LDH, morphology score and pH. PMID- 7496125 TI - [Suitability of monoclonal reagents for antigen determination in T-antigen activation]. AB - BACKGROUND: The lack of additional antibodies--for example anti-T--which can be contained in test sera of human origin has been pointed out as an advantage of monoclonal reagents in blood group serology. It was the aim of our study to examine whether the reactions of monoclonal reagents are nevertheless disturbed by T activation of red blood cells or not. MATERIALS AND METHODS: Monoclonal reagents of several manufacturers of the specificities anti-A, -B, -AB, -A1, -H, C, -c, -D, -E, -e, -K, -Jka, -Jkb, -Lea, -Leb, -M, and -N were tested. For this study we examined sialidase-treated and not treated red blood cells with and without the tested blood group antigen by the reagent using the tube centrifugation method. RESULTS: We found no significant disturbances for the monoclonal reagents of the AB0-system, A subgroups, Rhesus system, Kidd system, Kell antigen, and Leb antigen. Monoclonal anti-M and anti-N showed missing reactivity with sialidase-treated erythrocytes, which is already known from polyclonal test sera. Most of the monoclonal anti-Lea reagents showed strong false-positive reactions with T-positive Le(a-) erythrocytes. After several absorptions of one of the monoclonal anti-Lea reagents with T-activated Le(a-b-) red blood cells, the reactivity of the reagent with the Lea antigen and the T antigen had disappeared. CONCLUSIONS: In contrast to the other monoclonal reagents for most of the monoclonal anti-Lea reagents the lack of additional anti T antibodies does not indicate the lack of false-positive reactions. This cross reactivity might be caused by the fact that the type 1 chain antigen Lea and the type 3 chain antigen T have the same terminal saccharide (galactose) in beta 1- >3 connection to the preterminal saccharide of their peripheral core structure. PMID- 7496126 TI - [Batch documentation of blood products]. AB - BACKGROUND: In former years transmission of infectious diseases by blood products was repeatedly observed. To trace these incriminated batches of blood products proved to be nearly impossible. METHODS: This publication describes the documentation of different batches of blood products. In the simpliest case the batch numbers of blood products can be stored in files using forms. Besides this documentation of batches can be aided by EDP using databases and barcodes. A test for the correct and complete input of data is given. RESULTS: The registration of batches of commercially available blood products as well as blood preserves is possible. Incriminated batches can be traced back. CONCLUSIONS: Besides donor selection, control of the manufacturing process and the use of inactivation procedures for viruses the documentation of batches substantially contributes to the safety of plasma derivatives. With the applied techniques documentation is easily achieved. PMID- 7496127 TI - The study of glycoconjugates in pathological processes. PMID- 7496128 TI - Calnexin leads glycoproteins into the fold. PMID- 7496129 TI - Symposium to honour Lennart Roden glycosaminoglycans and connective tissue during the Rodenian era. PMID- 7496131 TI - Carbohydrate-based measurements on alpha-fetoprotein in the early diagnosis of hepatocellular carcinoma. AB - Serum alpha-fetoprotein (AFP) is a useful marker for the diagnosis of hepatocellular carcinoma (HCC), although this protein also increases moderately in benign liver diseases. The serum concentration of AFP in HCC at the time of initial diagnosis is now lower than before because of advancements in techniques for imaging the liver. The AFP concentration alone cannot distinguish between HCC and benign liver diseases, especially when it is less than 1000 ng ml-1. These circumstances lead to the need to discriminate between these diseases. This has been achieved by determining the carbohydrate structures of AFP by its reactivity with Lens culinaris agglutinin (LCA). The percentage of LCA-reactive species of AFP is significantly higher in HCC than in benign liver diseases. The fucosylation of the sugar chain at the innermost N-acetylglucosamine is the molecular basis of this variation. Therefore, the term 'fucosylation index' has been introduced to express the percentage of LCA-reactive species of AFP. This index is useful for the diagnosis of HCC even if the carcinoma is at an early stage. Furthermore, it can predict the development of HCC in the follow-up of chronic liver diseases. Thus, the qualitative and quantitative measurements of carbohydrate in AFP provide us with very valuable information for the differential diagnosis of various liver diseases. PMID- 7496130 TI - N-glycosylation/deglycosylation as a mechanism for the post-translational modification/remodification of proteins. PMID- 7496132 TI - Circulating blood group related carbohydrate antigens as tumour markers. AB - The various blood group related carbohydrate structures which are in clinical use as circulating tumour makers are reviewed. Their location on carbohydrate chains and their structural characteristics are shown, and their clinical performance in various malignant diseases is reviewed. The available data on their sensitivity, specificity and predictive value are shown; and carcinomas of the pancreas, ventricle, colon-rectum and ovary are identified as diseases in which these markers can be of good benefit for follow-up. Future research should be devoted to studies of the function of these structures, and to studies of their gene transcription. PMID- 7496133 TI - Evaluation of beta 1,4-galactosyltransferase in rheumatoid arthritis and its role in the glycosylation network associated with this disease. AB - Evidence indicating an important link between glycosylation changes and autoimmune rheumatic disease is presented. Attention is especially focused on the interrelationship between reduced galactosylation of the oligosaccharides of IgG, auto-sensitization which is thought to be of central importance in the pathogenesis of rheumatoid arthritis (RA), and the enzyme beta 1,4 galactosyltransferase (GTase) that catalyses the addition of galactose to the oligosaccharide chains on this molecule. Data are presented to indicate that GTase undergoes a variety of normal and disease associated changes. These variations are believed to contribute to the pathological processes in rheumatoid disease, and a hypothesis is suggested, whereby disease is associated with the dysregulation of an integrated glycosylation network, comprising IgG galactosylation, lymphocytic GTase and anti-GTase antibodies, that is a component of the normal immune system. PMID- 7496135 TI - Transferrin microheterogeneity as a probe in normal and disease states. AB - Isoelectric focusing of iron saturated serum has been established as a convenient method for showing transferrin glycan microheterogeneity. In a clinical setting, the method is used in the detection of cerebrospinal fluid leakage, the screening for surreptitious alcohol abuse and in the diagnosis of the carbohydrate deficient glycoprotein syndrome. In normal physiological states it can also be used as a tool to probe for changes in N-glycosylation. PMID- 7496134 TI - Glycosylation of alpha-1-proteinase inhibitor and haptoglobin in ovarian cancer: evidence for two different mechanisms. AB - The change in glycosylation of the two acute-phase proteins, alpha-1-proteinase inhibitor (API) and haptoglobin (Hp), in progressive ovarian cancer is different. This has been shown by monosaccharide analysis and lectin-binding studies of proteins purified from serum. In the glycan chains of API, there is decreased branching (more biantennary chains), less branches ending in alpha 2-3 sialic acid, more branches ending in alpha 2-6 sialic acid and more fucose, probably linked alpha 1-6 to the core region. On the other hand, Hp shows increased branching (more triantennary chains), more branches ending in alpha 2-3 sialic acid, less branches ending in alpha 2-6 sialic acid, and more fucose, probably in the alpha 1-3 linkage at the end of the chains. This is surprising because API and Hp are thought to be glycosylated by a common pathway in the liver. We have also shown that the fucose-specific lectin, lotus tetragonolobus, extracts abnormal forms of both Hp and API in ovarian cancer, but the expression of this Hp is related to tumour burden and the expression of this API is related to lack of response to therapy. It is suggested that this difference in the behaviour of API and Hp in ovarian cancer may be associated with the different changes in their glycosylation. Of the many mechanisms that could explain these findings, a likely one is that a pathological process is removing API with triantennary chains from the circulation.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7496137 TI - Changes of beta-1,4-N-acetylglucosaminyltransferase III (GnT-III) in patients with leukaemia. AB - Changes in the activity and transcription of UDP-N-acetylglucosamine: beta-D mannoside beta-1,4-N-acetylglucosaminyl-transferase III (GnT-III: EC 2.4.1.144) were investigated in haematological malignancies. GnT-III activity was elevated in patients with chronic myelogeneous leukaemia in blast crisis (CML-BC) and patients with multiple myeloma (MM); whereas most of the normal healthy subjects and patients with other haematological malignancies, including CML in its chronic phase, showed negligible activity. The GnT-III transcript of leukaemic cells from various haematological diseases showed a single band with a similar size. The ratio of GnT-III activity per normalized transcript in CML-BC was considerably higher than in the other conditions, which provided the possibility that in CML BC the transcript or the enzyme protein might be more stable, or that a post translational modification of the enzyme might enhance its activity. Furthermore, a lectin blot analysis of patient specimens and a lectin fluorescence study of CML cell lines revealed that E4-PHA binding to surface glycoproteins correlated with GnT-III activity, indicating that more bisecting GlcNAc was added to these glycoproteins, catalysed by elevated GnT-III in CML-BC. PMID- 7496136 TI - Alpha 1-acid glycoprotein (orosomucoid): pathophysiological changes in glycosylation in relation to its function. AB - The aim of this review is to summarize the research efforts of the last two decades with respect to (i) the determination and characterization of the changes in glycosylation of AGP under various physiological and pathological states; and (ii) the effects of such changes on its possible anti-inflammatory functions. It will become clear that the heterogeneity observed in the glycosylation of AGP in serum, represents various so-called glycoforms of AGP, of which the relative amounts are strictly determined by the (patho) physiological conditions. PMID- 7496138 TI - Glycoforms of serum alpha 1-acid glycoprotein as markers of inflammation and cancer. AB - alpha 1-acid glycoprotein (AGP) is a serum acute phase glycoprotein which possesses five N-linked complex type heteroglycan side chains which may be present as bi-, tri- and tetraantennary structures. Depending upon the content of biantennary structure on AGP, up to four glycoforms of AGP are present in serum. These glycoforms can be easily estimated in body fluids by means of crossed affinity-immunoelectrophoresis (CAIE) with the lectin, Concanavalin A (Con A). Con A selectively binds biantennary structures; the more biantennary structures on AGP, the stronger the binding. In acute inflammation, a relative increase of AGP glycoforms with biantennary units is observed-a type I glycosylation change. In some chronic inflammatory states there is an relative decrease of AGP glycoforms with biantennary heteroglycans-a type II glycosylation change. Moreover, in certain other states such as pregnancy, estrogen administration or liver damage, type II glycosylation changes are also seen. A detailed analysis of the clinical applications of the assessment of AGP glycoforms in sera of patients with rheumatic diseases, AIDS and various types of cancers is presented. Accumulated data shows that AGP glycoforms may be very useful in the detection of intercurrent infections in the course of rheumatoid arthritis, systemic lupus erythematosus, or myeloblastic leukaemia, and in the detection of secondary infections in human immunodeficiency virus infected individuals. AGP glycoforms are also very useful in differentiation between various forms of trophoblastic disease and are helpful in monitoring the treatment of these patients. Finally, AGP glycoforms provide valuable information for differentiation between primary and secondary liver cancer. PMID- 7496139 TI - Biochemical, histochemical and cell biological investigations on the actions of mistletoe lectins I, II and III with human breast cancer cell lines. AB - Cytotoxicity of the mistletoe lectins I, II and III towards six human breast cancer cell lines was assessed using the Mossman assay. In addition, binding of the three mistletoe lectins to the separated membrane glycoproteins of these cell lines, the binding and uptake of these lectins into the cells in tissue culture and the binding of the lectins to histological preparations of these cell lines were analysed. The results indicate that there are quantitative differences concerning the toxicity of these three lectins towards the different cell lines. Furthermore, the lectin binding pattern in the cell lines differed. In Western blots, several membrane glycoproteins were labelled by the lectins. Our results indicate subtle differences between the three lectins with regard to the parameters mentioned above; however, the toxicity of all three lectins from mistletoe is so similar that they all seem suitable for the construction of immunotoxins. PMID- 7496141 TI - The differences in structural specificity for recognition and binding between asialoglycoprotein receptors of liver and macrophages. AB - The Gal/GalNAc-specific lectin on the surface of rat peritoneal macrophages (macrophage asialoglycoprotein binding protein, M-ASGP-BP), which consists of a single polypeptide chain of 42 kDa, can form a homo-oligomeric receptor exhibiting high affinity for asialoorosomucoid (ASOR) [Ozaki K., Ii M., Itoh N., Kawasaki T. (1992) J Biol Chem 267: 9229-35]. In this study, the binding affinity of M-ASGP-BP was studied by using a series of synthetic or natural glycosides as inhibitors of 125I-ASOR binding to recombinant M-ASGP-BP expressed on COS-1 cells (rM-ASGP-BP), and the results were compared with those of human hepatic lectin (HHL) on Hep G2 cells. Clustering of multiple Gal (or GalNAc) residues increased the binding affinity to M-ASGP-BP as well as to HHL. In contrast to HHL and other mammalian hepatic lectins, rM-ASGP-BP bound Gal residues tighter than GalNAc residues. A galactose-terminated triantennary N-glycoside, having one N-acetyl lactosamine unit on the 6 branch and two N-acetyl-lactosamine units on the 3 branch of the trimannosyl core structure, showed affinity enhancement of approximately 10(5) over a monovalent ligand for HHL, while the same glycopeptide showed enhancement of about 2000-fold for rM-ASGP-BP. These results suggest that spatial arrangements of sugar combining sites and subunit organization of macrophage and hepatic lectins are different. PMID- 7496140 TI - Flow cytofluorimetric analysis of young and senescent human erythrocytes probed with lectins. Evidence that sialic acids control their life span. AB - Comparing the properties of 'young' and senescent ('aged') O+ erythrocytes isolated by applying ultracentrifugation in a self-forming Percoll gradient, we demonstrate that the sialic acids of membrane glycoconjugates control the life span of erythrocytes and that the desialylation of glycans is responsible for the clearance of the aged erythrocytes. This capture is mediated by a beta galactolectin present in the membrane of macrophages. The evidence supporting these conclusions is as follows: (1) Analysis by flow cytofluorimetry of the binding of fluorescein isothiocyanate labelled lectins specific for sialic acids shows that the aged erythrocytes bind less WGA, LPA, SNA and MAA than young erythrocytes. The binding of DSA and LCA is not modified. On the contrary, the number of binding sites of UEA-I specific for O antigen and of AAA decreases significantly. PNA and GNA do not bind to erythrocytes. (2) RCA120 as well as Erythrina cristagalli and Erythrina corallodendron lectins specific for terminal beta-galactose residues lead to unexpected and unexplained results with a decrease in the number of lectin binding sites associated with increasing desialylation. (3) The glycoconjugates from the old erythrocytes incorporate more sialic acid than the young cells. This observation results from the determination of the rate of transfer by alpha-2,6-sialyltransferase of fluorescent or radioactive N-acetylneuraminic acid, using as donors CMP-9-fluoresceinyl-NeuAc and CMP-[14C]-NeuAc, respectively. (4) Microscopy shows that the old erythrocytes are captured preferentially by the macrophages relative to the young ones. Fixation of erythrocytes by the macrophage membrane is inhibited by lactose, thus demonstrating the involvement of a terminal beta-galactose specific macrophage lectin. (5) Comparative study of the binding of WGA, LPA, SNA and MAA to the aged erythrocytes and to the in vitro enzymatically desialylated erythrocytes shows that the desialylation rate of aged cells is low but sufficient to lead to their capture by the macrophages. PMID- 7496142 TI - Versatile synthesis of bi- and tri-antennary galactose ligands: interaction with the Gal/GalNAc receptor of human hepatoma cells. AB - We have synthesized bi- and tri-antennary galactose ligands by coupling 1-thio beta-D-galactose derivatives to the alpha- and epsilon-amino groups of L-lysine and L-lysyl-L-lysine via highly flexible hydrophilic spacer arms that allow variation of their intergalactose distances. The interaction of these ligands with the Gal/GalNAc receptor of HepG2 cells showed a binding affinity that was: (i) in agreement with the clustering effect known to occur with more complex oligomeric structures, i.e. tri- > bi-antennary; ii) dependent on the intergalactose distances (optimal interactions were observed for the tri antennary structures with distances > 2 nm). These ligands, that can be easily conjugated to bioactive (macro) molecule carrier systems, could be useful for their targeting to hepatocytes. PMID- 7496143 TI - FAB CID-MS/MS characterization of tetrasaccharide tri- and tetrasulfate derived from the antigenic determinant recognized by the anti-chondroitin sulfate monoclonal antibody MO-225. PMID- 7496144 TI - Characterization of four monosialo and a novel disialo Asn N-glycosides from the urine of a patient with aspartylglycosaminuria. AB - We previously reported for the first time two Japanese patients with aspartylglycosaminuria (AGU). A novel disialo Asn N-glycoside (AG-5) has been isolated from the urine of one of the patients in addition to four known monosialo Asn N-glycosides (AG-1 to AG-4) by gel filtration and anion exchange chromatography in this study. Final purification of AG-5 was achieved by an electrochemical chromatographic method, high performance liquid chromatography with pulsed amperometric detector (HPLC-PAD). The yield of AG-5 was approximately 1 mg l-1 urine. The chemical structures of AG-1 to AG-5 were characterized by gas liquid chromatography, a permethylation study, fast atom bombardment-mass spectrometry (FAB-MS), and nuclear magnetic resonance (NMR). Based on the structural analysis, AG-5 had the following novel structure: NeuAc alpha 2- >8NeuAc alpha 2-->3Gal beta 1-->4GlcNAc beta 1-->Asn. PMID- 7496145 TI - Lipooligosaccharidic antigens from Mycobacterium kansasii and Mycobacterium gastri. AB - A set of Lipooligosaccharides (LOSs) has previously been characterized in M. gastri W471. The structure of the highly antigenic LOS (LOS-III) was elucidated and this molecule can unambiguously distinguish M. gastri from the opportunistic pathogen M. kansasii. In the present study, the structures of three other M. gastri W471 LOSs were determined by one-dimensional 1H-NMR spectroscopy and gas liquid chromatography. They differ by the number of Xylp units and by the structure of the distal monosaccharide. The two dimensional (2D) NMR approach was successfully applied to the LOS antigen of M. kansasii to locate the acetyl and acyl substituents and to determine the anomeric configuration of the alpha-D-Fucp unit. The molecular specificity of anti-LOS-III antibodies was investigated and the LOS-III epitope was defined as the distal disaccharide: 3,6-dideoxy-4-C-(1,3 dimethoxy-4,5,6,7-tetrahydroxy- heptyl)-alpha-xylohexp-(1-->3)-beta-L-Xylp. PMID- 7496146 TI - Structural and immunochemical identification of Leb glycolipids in the plasma of a group O Le(a-b-) secretor. AB - Total non-acid glycosphingolipids were isolated from the plasma of a healthy red blood cell group O Le(a-b-) salivary ABH secretor individual. Glycolipids were fractionated by HPLC and combined into eight fractions based on chromatographic and immunoreactive properties. These glycolipid fractions were analysed by thin layer chromatography and tested for Lewis activity with antibodies reactive to the type 1 precursor (Le(c)), H type 1 (Le(d)), Le(a) and Le(b) epitopes. Fractions were structurally characterized by mass spectrometry (EI-MS and LSIMS) and proton NMR spectroscopy. Expected blood group glycolipids, such as H type 1, (Fuc alpha 1-2Gal beta 1-3GlcNac beta 1-3Gal beta 1-4Glc beta 1-1Cer) were immunochemically and structurally identified. Inconsistent with the red cell phenotype and for the first time, small quantities of Le(b) blood group glycolipids (Fuc alpha 1-2Gal beta 1-3(Fuc alpha 1-4)GlcNAc beta 1-4Glc beta 1 1Cer) were immunochemically and structurally identified in the plasma of a Lewis negative individual. These findings confirm recent immunological evidence suggesting the production of small amounts of Lewis antigens by Lewis negative individuals. PMID- 7496147 TI - Variation in N-linked carbohydrate chains in different batches of two chimeric monoclonal IgG1 antibodies produced by different murine SP2/0 transfectoma cell subclones. AB - Two chimeric human/murine monoclonal antibodies were constructed by substitution of the murine constant regions with human gamma 1 and kappa constant regions for heavy and light chains, respectively. The chimeric human/murine molecules are anti-idiotypic antibodies, meaning that they were directed against the antigen binding site in the variable region of another antibody. Antibody batches were produced under identical production conditions, using two selected SP2/0 myeloma cell subclones, which produce chimeric antibodies with different variable regions, but identical constant regions. Several samples were collected during the production of the antibodies in hollow-fibre reactors. The heavy chain, but not the light chain, of the two different chimeric IgG1 antibodies is glycosylated. Structural analysis of the enzymically released N-linked carbohydrate chains by 1H-NMR spectroscopy, as well as by chromatographic profiling, demonstrated that the collection of N-glycans comprises a small amount of monoantennary, and for the greater part diantennary structures. The N-glycans are completely (alpha 1-->6)-fucosylated at the innermost GlcNAc residue. The antennae of the neutral diantennary N-glycans are built up from GlcNAc beta 1- >2, Gal beta 1-->4GlcNAc beta 1-->2 or Gal alpha 1-->3G alpha 1 beta 1-->4GlcNAc beta 1-->2 elements, whereas the antennae of the neutral monoantennary carbohydrate chains have only (beta 1-->2)-linked GlcNAc residues. Galactosylation of the GlcNAc beta 1-->2Man alpha 1-->6 branch occurs four times more frequently than that of the GlcNAc beta 1-->2Man alpha 1-->3 branch, independently of the production batch. A small amount of the diantennary N glycans are mono- or disialylated, carrying N-acetylneuraminic acid (Neu5Ac) or N glycolylneuraminic acid (Neu5Gc), exclusively (alpha 2-->6)-linked to beta Gal. Analysis of the different production batches demonstrates that the structures of the N-linked carbohydrate chains are identical in the two chimeric antibodies, but that the relative amounts of the major oligosaccharide components, the degree of sialylation and the molar ratio of Neu5Ac to Neu5Gc varies with the SP2/0 cell subclone, and only slightly with cell age. PMID- 7496148 TI - Computer simulation of histo-blood group oligosaccharides: energy maps of all constituting disaccharides and potential energy surfaces of 14 ABH and Lewis carbohydrate antigens. AB - The three-dimensional structures of fourteen histo-blood groups carbohydrate antigens have been established through a combination of molecular mechanics and conformational searching methods. The conformational space available for each disaccharide, constituents of these determinants, has been throroughly characterized. The results have been organized in a data bank fashion. Larger relatives, i.e. 14 tri- and tetrasaccharides of histo-blood group antigens, have been modelled using a different method for exploring the complex potential energy surface. This approach is aimed at establishing all the possible families of conformations, along with the conformational pathways. Different conformational behaviours are exhibited by these oligosaccharides. Some of them, i.e. Le(x) and Le(y) tri and tetrasaccharides, are very rigid; 99% of their populations belong to the same conformational family. Others, like H type 1, H type 2 or H type 6 oligosaccharides, are essentially rigid, but a secondary conformational family, corresponding to 3-4% of the total population, can arise. Finally, the H types 3 and 4 trisaccharides, and the A type 1 and A type 2 tetrasaccharides are predicted to behave rather flexibly. The information gathered in the present investigation has been used to analyse the body of experimental evidence, either physical or biological, available for this series of carbohydrate antigens. Of special interest are the several different alignments that can be proposed for these molecules. They yield a realistic definition of the three-dimensional features of the epitopes thereby providing essential information about how carbohydrate antigens are recognized by proteins. PMID- 7496149 TI - Accessible hyaluronan receptors identical to ICAM-1 in mouse mast-cell tumours. AB - Immunohistochemical studies of the hyaluronan (HA)-receptor (R), originally found on liver endothelial cells (LEC) and related to the intercellular adhesion molecule 1 (ICAM-1), showed that polyclonal antibodies against HARLEC (HA receptor on LEC) also stain structures in mouse mastocytomas, mainly vessels. To test if intravenously administered HA might target the tumour receptors in vivo, mice carrying an inoculated mastocytoma in one hind leg muscle were injected in the tail vein with 125I-tyrosine (T)-labelled HA and killed 75 min after injection when organs and tissues were checked for radioactivity. When doses exceeding the binding capacity of the liver were injected, a significant increase in radioactivity (up to five-fold) within the tumour tissue was found. The weight adjusted difference between control and tumour tissue was greater for smaller tumours, probably due to necrosis in the larger. HA-staining of tumours from animals receiving 125I-T-HA, showed HA in areas that also stained weakly for ICAM 1 using monoclonal antibodies. ICAM-1 staining was dramatically increased after hyaluronidase treatment of the sections, indicating that the HA is bound to these receptors and thereby blocks antibody recognition. PMID- 7496150 TI - Expression of the T antigen on a T-lymphoid cell line, supT1. AB - We have measured glycosyltransferase activities of SupT1 cells, a T-lymphoid cell line shown to react with autoantibodies in the sera of many HIV patients. Since considerable alpha-N-acetylgalactosaminyl-transferase and beta 1, 3 galactosyltransferase activities were found in SupT1 cells, at least the O-glycan core 1 structure can probably be synthesized. FACS analysis using an anti-T monoclonal antibody showed expression of the T antigen (Gal beta 1-3 GalNAc). Glycoproteins with the T antigen were isolated by immunoprecipitation with the anti-T antibody from a SupT1 cell lysate labelled metabolically with 3H glucosamine and then analysed by SDS-PAGE. It was revealed that the precipitate contained a glycoprotein with a molecular weight corresponding to that of leukosialin. O-glycans were prepared from the immunoprecipitate by alkaline borohydride treatment and then fractionated on Bio-Gel P-2, GalNAcOH and Gal GalNAcOH being identified inter alia. These results suggest that an anti-T antibody may be included in the autoantibodies found in HIV-1 infected individuals. PMID- 7496151 TI - Insertion into Aspergillus nidulans of functional UDP-GlcNAc: alpha 3-D- mannoside beta-1,2-N-acetylglucosaminyl-transferase I, the enzyme catalysing the first committed step from oligomannose to hybrid and complex N-glycans. AB - Filamentous fungi are capable of secreting relatively large amounts of heterologous recombinant proteins. Recombinant human glycoproteins expressed in this system, however, carry only carbohydrates of the oligomannose type limiting their potential use in humans. One approach to the problem is genetic engineering of the fungal host to permit production of complex and hybrid N-glycans. UDP GlcNAc:alpha 3-D-mannoside beta- 1,2-N-acetylglucosaminyltransferase I (GnT I) is essential for the conversion of oligomannose to hybrid and complex N-glycans in higher eukaryotic cells. Since GnT I is not produced by fungi, we have introduced into the genome of Aspergillus nidulans the gene encoding full-length rabbit GnT I and demonstrated the expression of GnT I enzyme activity at levels appreciably higher than occurs in most mammalian tissues. All the GnT I activity in the Aspergillus transformants remains intracellular suggesting that the rabbit trans membrane sequence may be capable of targeting GnT I to the fungal Golgi apparatus. PMID- 7496152 TI - Substrate specificity and inhibition of UDP-GlcNAc:GlcNAc beta 1-2Man alpha 1-6R beta 1,6-N-acetylglucosaminyltransferase V using synthetic substrate analogues. AB - UDP-GlcNAc:GlcNAc beta 1-2Man alpha 1-6R (GlcNAc to Man) beta 1,6- N acetylglucosaminyltransferase V (GlcNAc-T V) adds a GlcNAc beta 1-6 branch to bi- and triantennary N-glycans. An increase in this activity has been associated with cellular transformation, metastasis and differentiation. We have used synthetic substrate analogues to study the substrate specificity and inhibition of the partially purified enzyme from hamster kidney and of extracts from hen oviduct membranes and acute myeloid leukaemia leukocytes. All compounds with the minimum structure GlcNAc beta 1-2Man alpha 1-6Glc/Man beta-R were good substrates for GlcNAc-T V. The presence of structural elements other than the minimum trisaccharide structure affected GlcNAc-T V activity without being an absolute requirement for activity. Substrates with a biantennary structure were preferred over linear fragments of biantennary structures. Kinetic analysis showed that the 3-hydroxyl of the Man alpha 1-3 residue and the 4-hydroxyl of the Man beta- residue of the Man alpha 1-6(Man alpha 1-3)Man beta-R N-glycan core are not essential for catalysis but influence substrate binding. GlcNAc beta 1-2(4,6-di-O methyl-)Man alpha 1-6Glc beta-pnp was found to be an inhibitor of GlcNAc-T V from hamster kidney, hen oviduct microsomes and acute and chronic myeloid leukaemia leukocytes. PMID- 7496154 TI - Autotaxin is an N-linked glycoprotein but the sugar moieties are not needed for its stimulation of cellular motility. AB - Autotaxin is a 125kD autocrine motility factor that stimulates both random and directed motility in producing the human A2058 melanoma cell line. The recently cloned autotaxin has been demonstrated to bind strongly and specifically to concanavalin A (con A). In this study, we show that the oligosaccharide side chains on autotaxin are exclusively asparagine linked, since N-glycosidase F, but not neuraminidase or O-glycosidase, decreases the protein molecular mass to 100 105kD, which is the calculated molecular mass of the deduced autotaxin polypeptide. Furthermore, removal of oligosaccharide side chains by N-glycosidase F can be performed under mild conditions that retain motility-stimulating activity, suggesting that the oligosaccharide side chains are not necessary for autotaxin to activate its receptor. Finally, when melanoma cells are treated with inhibitors of carbohydrate processing, such as N-methyl-1-deoxynojirimycin, 1 deoxymannojirimycin and swainsonine, they still secrete a motility-stimulating autotaxin. Therefore, the carbohydrate side chains on autotaxin are not necessary to stimulate motility; however, they may still play a role in folding, secretion or maintenance of the active conformation of the protein. PMID- 7496153 TI - Glycan modification of a thermostable recombinant (1-3, 1-4)-beta-glucanase secreted from Saccharomyces cerevisiae is determined by strain and culture conditions. AB - High level biosynthesis and secretion of the thermostable hybrid (1-3, 1-4)- beta glucanase H(A16-M) has been achieved in Saccharomyces cerevisiae by means of the yeast vacuolar endoprotease B promoter (PRB1P) and the Bacillus macerans (1-3, 1 4)-beta-glucanase signal peptide. The N-glycans present on the yeast-secreted H(A16-M), denoted H(A16-M)-Y, were released by endoglycosidase H, and identified by proton NMR spectroscopy to be a homologous series of Man8-13GlcNAc2, although only traces of Man9GlcNAc2 were found. Therefore, processing of N-glycans on H(A16-M)-Y is similar to that on homologous proteins. Most of the N-glycans (88%) were neutral while the remainder were charged due to phosphorylation. Site directed mutagenesis of Asn to Gln in two of the N-glycosylation sequons, and subsequent analysis of the N-glycans on the yeast-secreted proteins together with analysis of the N-glycans from the individual sites of H(A16-M)-Y suggest the presence of steric hindrance to glycan modification by the glycans themselves. H(A16-M)-Y produced under control of either the yeast protease B or the yeast 3' phosphoglycerate kinase promoter, each in two different Saccharomyces strains revealed a dependence of N-glycan profile on both strain and culture conditions. The extent of O-glycosylation was found to be nine mannose units per H(A16-M)-Y molecule. An attempt to identify the linkage-sites for the O-glycans by amino acid sequencing failed, suggesting non-stoichiometric or heterogeneous O glycosylation. The possible modes in which N-glycans might contribute to resistance of H(A16-M)-Y to irreversible thermal denaturation are discussed with respect to structural information available for H(A16-M)-Y. PMID- 7496155 TI - In vivo autofluorescence of an unpigmented melanoma in mice. Correlation of spectroscopic properties to microscopic structure. AB - Recently, fluorescence spectroscopy and imaging have been under investigation for in vivo diagnosis of several types of superficial cancer, including primary melanomas of the skin. Here we report on a detailed investigation of the autofluorescence properties of a K1735P melanoma implanted intradermally in the ears of syngeneic C3H/HeN mice. Although this tumour can produce melanin in some cases, it appeared as an unpigmented lesion in our experiments. Excitation emission maps in the wavelength range of 360-700 nm were recorded from tumours and normal ears. The control ears and the tumour-bearing ears showed fluorescence in a broad range of excitation and emission wavelengths. Valleys of decreased fluorescence were observed in the 385-425 nm range and could be related to absorption of the excitation light by haemoglobin, oxyhaemoglobin and a third unknown absorber. The spectroscopic differences between the malignant melanoma and the control skin could be related to either differences in blood oxygenation or the tissue dimensions. However, no spectroscopic features were detected reflecting intrinsic differences between the melanoma and normal tissue. PMID- 7496156 TI - Immunization with interleukin-2/interferon-gamma double cytokine-secreting allogeneic fibroblasts prolongs the survival of mice with melanoma. AB - LM mouse fibroblasts (H-2k) were modified for the expression of (antibody defined) melanoma-associated antigens (MAA) and the secretion of interleukin-2 (IL-2) and interferon-gamma (IFN-gamma) (RLBA-IL-2/IFN-gamma cells). The cell construct was tested for its immunogenic properties in C57BL/6 mice (H-2b) with B16 melanoma. The results indicated that the survival of mice injected with a mixture of B16 cells and the modified, double cytokine-secreting fibroblasts was significantly longer than that of mice injected with B16 cells and LM cells modified for the expression of MAA and the secretion of IL-2 or IFN-gamma alone (RLBA-IL-2 or RLBA-IFN-gamma cells). Both natural killer/lymphokine-activated killer (NK/LAK) cells and Lyt-2.2 + CTLs with anti-melanoma cytotoxic activities were predominant in mice immunized with the double cytokine-secreting cells. B16 melanoma cells persisted in mice treated with RLBA-IL-2 cells (B16-R3). The B16 R3 cells were resistant to anti-melanoma effector cells from mice immunized with RLBA-IL-2 cells. The recurrent melanoma cells were deficient in the expression of MHC class I determinants. Class I expression by B16-R3 cells was increased if they were incubated in medium conditioned by the growth of IFN-gamma-secreting RLBA-IL-2/IFN-gamma or RLBA-IFN-gamma cells. After incubation, the sensitivity of B16-R3 melanoma cells to immune-effector cells from mice immunized with RLBA-IL-2 cells was restored. The survival of mice bearing low MHC class I-expressing B16 R3 cells, treated RLBA-IL-2/IFN-gamma cells, was determined. The treated animals survived significantly longer than mice with B16-R3 melanoma treated with RLBA-IL 2 cells. Similar results were obtained for mice with B16-R3 melanoma treated with RLBA-IFN-gamma cells. We postulate that immunization of mice with IL-2/IFN-gamma double cytokine-secreting cells stimulated multiple anti-melanoma effector mechanisms. Analogous to the enhanced therapeutic anti-tumour effects of combination chemotherapy, it was likely that treatment with a cellular immunogen engineered to stimulate more than one effector mechanism resulted in the elimination of larger numbers of tumour cells than treatment with an immunogen that stimulated a single effector mechanism alone. PMID- 7496157 TI - Stepdown hyperthermia in human melanoma cells: effects on protracted mild hyperthermia for survival and DNA polymerase inactivation. AB - Human melanoma cells have been used to evaluate whether stepdown heating (SDH) could increase the effectiveness of long-duration mild hyperthermia (LDMH). The effects of these treatments were also evaluated on cell survival and DNA polymerase inactivation. Short treatments (30 min) at 43 degrees C did not result in much SDH effect for subsequent protracted heating at 40 degrees C. The effect on DNA polymerases was also very small. However, heating at 44 degrees C for 30 min had a large SDH heating effect on subsequent heating at 40 degrees C and 41 degrees C. The SDH effect occurred mainly in the first 5-10 h of subsequent LDMH and, at longer heating times, the rate of cell killing was reduced. The 44 degrees C SDH effect was also observed on DNA polymerase inactivation. Comparing the degree of cell killing and polymerase inactivation showed a good correlation for the various SDH protocols. PMID- 7496158 TI - Cytokine-mediated modulation of integrin, ICAM-1 and CD44 expression on human uveal melanoma cells in vitro. AB - Adhesion molecules are likely to play a role in the process of tumour progression. We investigated the expression of integrins, ICAM-1, and CD44 and the influence of interferon-alpha (IFN-alpha), interferon-gamma (IFN-gamma), and tumour necrosis factor-alpha (TNF-alpha) on expression of these molecules on four uveal melanoma cell lines. The in vitro integrin expression was quite variable. The alpha V and beta 1 subunits were expressed on all cell lines, and none of the cell lines showed any alpha 3, beta 2, or beta 4 expression. Other integrin subunits showed a more variable pattern. ICAM-1 and CD44 were strongly expressed on all cell lines. IFN-alpha, IFN-gamma, and TNF-alpha upregulated alpha 1, alpha 2, and alpha 3 expression, and did not alter alpha 4, alpha 5, alpha 6, beta 2, alpha v beta 3, and beta 4 expression. The effects on alpha V and alpha V beta 5 were variable. ICAM-1 was upregulated by IFN-gamma and TNF-alpha, but not by IFN alpha. Cytokine treatment hardly changed CD44 expression. In one case a comparison was made between expression on cultured cells and on tissue sections of the tumour of origin. Differences in expression were observed for the integrin subunits alpha 2, alpha 3, and alpha 5. This study shows that integrins and ICAM 1 expression on uveal melanoma cells in vitro are susceptible to cytokine treatment, but that the effects on integrin expression are cytokine and cell line dependent. Furthermore, some differences in integrin expression between cells in vivo and in vitro exist. PMID- 7496159 TI - Mutations and defective expression of the WAF1 p21 tumour-suppressor gene in malignant melanomas. AB - The WAF1 gene, located on chromosome 6p, encodes a M(r) 21,000 protein (p21) that can arrest cell growth by associating with and inhibiting cyclin-dependent kinase complexes that are necessary for cells to exit Gr. Transcriptional activation of WAF1 can be accomplished by increasing levels of p53 protein induced by various cellular stresses, including DNA damage. Metastatic melanomas are paradoxical in that most overexpress wild-type p53 protein, yet cell growth is not inhibited. Thus, it is possible that lack of growth suppression in melanomas is due, in part, to mutations in the WAF1 gene. Therefore, we examined the entire coding region of the WAF1 gene in 24 metastatic melanoma cell lines and three normal melanocyte lines by single-strand conformation polymorphism (SSCP) analysis and direct DNA sequencing. We similarly examined the DNA from lymphoblastoid cell lines, derived from nine individuals belonging to seven melanoma-prone families, in which haplotypes of markers on 6p cosegregate with melanoma for germline mutations in the WAF1 gene. Results indicate that (i) mutation of the WAF1 gene is an infrequent event in individuals with sporadic melanoma or predisposed to familial melanoma and (ii) the uncontrolled growth of melanoma cells is not due to mutation of the WAF1 gene. However, expression studies found a wide variation in the level of p21 protein in melanoma cells, suggesting that aberrant regulation of p21 may play a role in melanoma development. Moreover, there was no predictable relationship between p21 expression and p53 expression, indicating that other, p53-independent, pathways may be important for the regulation of p21 in melanoma cells. PMID- 7496160 TI - TAL1 gene deletions and TAL1 protein expression in sporadic melanoma. AB - Studies on cytogenetic abnormalities and cell lines have implicated chromosome 1p32 as being important in the pathogenesis of melanoma. Genetic linkage studies have also mapped a melanoma-susceptibility locus to chromosome 1p. The gene TAL1 is present on chromosome 1p32, and deletions within it are the commonest chromosomal abnormality in T-acute lymphoblastic leukaemia (T-ALL). A melanoma cell line harbouring a 1p32 deletion involving the TAL1 gene and the presence of TAL1 protein in developing mouse melanocytes led us to investigate whether TAL1 deletions and/or TAL1 protein expression occur in sporadic melanomas. DNA extracted from 32 fresh melanomas was amplified by standard polymerase chain reaction for the four common deletions of the TAL1 gene that occur in T-ALL. In addition, frozen and paraffin-embedded sections of these melanomas were stained with monoclonal antibodies that detect full-length and truncated TAL1 protein. The results of the study show that deletions of TAL1 do not occur in melanoma. Indeed, full and truncated TAL1 protein also could not be detected immunohistochemically in the paraffin-embedded and frozen sections of the melanomas. We conclude that the TAL1 gene and its protein are probably not directly involved in the oncogenesis of melanomas. PMID- 7496162 TI - Sinonasal malignant melanoma--a clinicopathologic analysis of 18 cases. AB - Sinonasal melanoma is a rare malignancy. We present the clinicopathologic review of 18 cases seen at the British Columbia Cancer Agency between 1976 and 1992: 13 men and five women, mean age 66 years (range 32-88). Patients presented with nasal obstruction and bleeding (n = 8), obstruction alone (n = 4), bleeding alone (n = 5) or pain (n = 1). Those with bleeding presented with a shorter duration of symptoms than those with obstruction alone. All patients with obstruction alone died of their disease, while all patients with bleeding alone are alive or have died of an unrelated cause; four out of eight patients with both obstruction and bleeding are alive. There was no significant relationship between treatment modality and outcome. Histologic subtypes included epithelioid (n = 10), spindle cell (n = 4), small-cell (n = 3) and pleomorphic (n = 1). Eight out of 11 cases from whom samples of paraffin-embedded tissue were available showed more prominent staining for HMB-45 than for S-100. In two cases, only rare (< 0.1%) cells stained for S-100. Cell type, mitotic rate and P53 expression were unrelated to disease outcome. Six out of seven patients with < or = 10% of cells showing intense staining for PCNA were alive or had died of an unrelated cause, while three out of four with > 10% staining died of their disease.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7496161 TI - Sentinel lymph node status as an indicator of the presence of metastatic melanoma in regional lymph nodes. AB - The value of elective lymph node dissection (ELND) for melanoma patients with clinically uninvolved regional nodes remains controversial. However, it has been proposed that selective 'sentinel' lymph node biopsy reliably identifies individuals with micrometastases, who are most likely to benefit from full ELND. The aim of this study was to confirm that metastatic melanoma cells travelling in lymphatics do not bypass the sentinel node. After preoperative lymphoscintigraphy and intraoperative injection of blue dye around the primary melanoma site, sentinel node biopsy was performed in 118 melanoma patients for whom full ELND was planned as part of their definitive surgical treatment. A confidently identified sentinel node was tumour positive in 22 out of 105 regional lymph node fields (21%). In 18 cases the sentinel node was the only node found to be involved and in four cases, additional nodes were positive. In two other patients a positive node was found when the sentinel lymph node had been negative. However, in each case an avoidable error of technique had occurred and definite blue staining indicated that the positive node was in fact another sentinel node. This study thus confirms that sentinel lymph node status reliably indicates whether metastatic melanoma is present in regional lymph nodes. PMID- 7496163 TI - The histological and immunohistochemical changes in the skin of patients with melanoma who develop changes in skin pigmentation following immunotherapy. AB - The histological and immunocytochemical appearances of skin with altered pigmentation from two patients receiving chemoimmunotherapy for melanoma were examined. In both patients, there was a clinical response to treatment coincident with depigmentation of skin and hair. Skin biopsies showed extensive infiltration with CD8+ and CD4+ lymphocytes rather than CD57+ in the depigmented areas suggestive of a specific MHC-related cytotoxic T-cell activity against melanocytes. In keeping with this, MHCII expression was markedly up-regulated. These observations suggest the development of a simultaneous anti-tumour and anti melanocyte immune response stimulated by chemoimmunotherapy, possibly against the same or similar antigens. PMID- 7496164 TI - Phase II trial of recombinant interferon-alpha with BCNU, cisplatin, DTIC and tamoxifen in advanced malignant melanoma. AB - Since cytotoxic chemotherapy (BCNU, DTIC and cisplatin, tamoxifen) and interferon alpha (IFN-alpha) have each produced responses in advanced malignant melanoma, a phase II trial was conducted to evaluate the response and toxicity of simultaneous administration of both therapies. Of 33 assessable patients, two (6%) had complete response (CR) and 12 patients (36%) had partial response (PR), for a total response rate (CR+PR) of 42% (95% confidence interval 26-58). Four patients had minor response (12%). Mixed responses occurred in five patients (15%). The remaining patients had progressive disease. The duration of CR was 3, 7 and 17 (+) months and the duration of PR was 3+ to 19+ months (median 6 months). The median overall survival for all patients entered into the study was 5 months. Main toxicities included myelosuppression and fatigue. Combined simultaneous cytotoxic chemotherapy and IFN produced a high response rate (42%) which is comparable to that reported for chemotherapy alone. Further studies are needed to determine the optimal schedule for combining chemotherapy and immunotherapeutic agents as well as the impact of biological agents on survival in the treatment of melanoma. PMID- 7496165 TI - Dacarbazine and interferon-alpha 2a in advanced malignant melanoma: high response rate and prolongation of response duration occur in different patient subpopulations. AB - Fifty-three evaluable patients with metastatic malignant melanoma were enrolled in a phase II prospective study designed to assess the response rate, time to progression and survival after dacarbazine (DTIC) and interferon-alpha 2a (IFN alpha 2a) treatment in patients with local metastatic disease compared with patients with distant metastases. Patients received intravenous DTIC from day 1 at a dosage of 400-500 mg/m2, repeated every 21 days (in the case of good tolerance--25 patients--the dose was increased to 600-800 mg/m2) combined with subcutaneous IFN-alpha 2a (9 x 10(6) U three times/week, increased in the case of good tolerance to 15 x 10(6) U three times/week). Forty-two patients with distant metastases were compared with 11 patients who had local metastatic disease. Three complete (6%) and six partial (11%) responses were seen, with an overall response rate of 17% (95% confidence interval 8-29). Patients with local metastases had a higher response rate (50%) compared with patients with distant metastases (visceral involvement, mediastinal and para-aortic lymph node metastases) (10%; p = 0.01). The median overall survival was 4.5 months. The progression-free interval for responders with distant metastases was significantly longer (11 months), than for responders with local metastases (4.5 months) (p = 0.004). These results may suggest that the combination treatment DTIC/IFN-alpha has a greater benefit in terms of longer progression-free interval in responders with distant metastases. PMID- 7496167 TI - Neuromuscular disorders: gene location. PMID- 7496166 TI - Addition of dacarbazine or cisplatin to interferon-alpha/interleukin-2 in metastatic melanoma: toxicity and immunological effects. AB - The combination of chemotherapy and immunotherapy seems to improve response rate in metastatic melanoma. We investigated the effects on toxicity and immunological effects of a single dose of dacarbacin (DTIC; 850 mg/m2) or cisplatin (CDDP; 100 mg/m2) added to subsequent immunotherapy with interferon-alpha (IFN-alpha) and interleukin-2 (IL-2). Twelve patients, who did not respond to IFN-alpha/IL-2 alone were studied. Six received DTIC and IFN-alpha/IL-2, and six received CDDP and IFN-alpha/IL-2. DTIC did not add significant toxicity except for nausea. Significant thrombocytopenia was observed in two patients after CDDP. Although CDDP led to grade 3 nephrotoxicity in two patients, the IL-2-induced fluid retention was less severe than with IFN-alpha/IL-2 alone. Pharmacokinetics of IL 2 were not altered by DTIC, but higher IL-2 serum levels were found in patients with grade 3 nephrotoxicity after CDDP. The IL-2-related induction of secondary mediators (interferon-gamma, tumour necrosis factor-alpha, soluble CD25) was not impaired by chemotherapy and the induction of neopterin was significantly higher after addition of CDDP. One partial response was observed after addition of DTIC to IFN-alpha/IL-2, and one after addition of CDDP. The addition of a single dose of DTIC or CDDP to IFN-alpha/IL-2 is fairly well tolerated and does not abolish induction of secondary mediators. Randomized trials are necessary to test the clinical efficacy. PMID- 7496168 TI - Mitochondrial encephalomyopathies: gene mutation. PMID- 7496169 TI - Identification of three novel mutations in the gene for Cu/Zn superoxide dismutase in patients with familial amyotrophic lateral sclerosis. AB - About 10% of cases of amyotrophic lateral sclerosis (ALS), a paralytic disorder characterized by death of motor neurons in the brain and spinal cord, exhibit autosomal dominant inheritance. A subgroup of these familial cases are caused by mutations in the gene encoding Cu/Zn superoxide dismutase (SOD1). We report here three additional mutations occurring in the SOD1 gene in three families with ALS. Two of these changes are missense mutations in exon 5 of the SOD1 gene, resulting in leucine 144 to serine and alanine 145 to threonine substitutions. The third, a single base pair change in intron 4 immediately upstream of exon 5, results in an alternatively spliced mRNA. The alternate transcript conserves the open reading frame of exon 5, producing an SOD1 protein with three amino acids inserted between exons 4 and 5 (following residue 118). These three mutations bring to 29 the total number of distinct SOD1 mutations associated with familial ALS. PMID- 7496170 TI - Possible strategies for treatment of SMA patients: a neurobiologist's view. AB - This paper discusses possible strategies that might prevent or alleviate muscle weakness of SMA patients and hence improve their condition. The strategies discussed are as follows. (1) Prevention of motoneurone death. To achieve this two main approaches have been applied. Firstly, trophic factors have been used to prevent motoneurone death after nerve injury and clinically in diseases such as motoneurone disease. The results of these attempts will be described. Secondly, the possibility that injured motoneurones die as a result of the excitotoxic effects of the excitatory transmitter glutamate will be explored. Evidence will be presented which indicates that blocking glutamate receptors can rescue injured motoneurones from death. (2) Replacement of lost motoneurones by embryonic grafts. Motoneurones from grafts of embryonic spinal cord have been shown to survive in the adult spinal cord and are able to reinnervate skeletal muscles. The potential and practical problems of this approach will be discussed. (3) Expansion or motor unit territory of surviving motoneurones. Such an expansion of the territory occupied by individual motor units can be achieved by encouraging sprouting and ensuring that the newly formed connections between the motoneurone and muscle fibres are maintained, so that individual motor units are capable of developing more force. Strategies to achieve such an expansion of motor unit territory will be described. Finally, combinations of some of these approaches are considered. PMID- 7496172 TI - Lecithinized superoxide dismutase retards wobbler mouse motoneuron disease. AB - Gene mutations of Cu/Zn superoxide dismutase (SOD) have been discovered in familial amyotrophic lateral sclerosis (ALS). Oxidative stress also plays a role in the pathogenesis of sporadic ALS. Whether antioxidant therapy is beneficial in this fatal disease is now crucial. We have shown that SOD treatment improves neuromuscular dysfunction and morphological changes in wobbler mouse motoneuron disease. Progressive spinal motor neuronopathy and axonopathy, predominantly in the cervical cord, occur at postnatal age 3-4 weeks, leading to muscle weakness and contracture of the forelimbs in this animal. These motor deficits rapidly increase by postnatal age 6-8 weeks, and then slowly progress. Wobbler mice were given two doses daily of phosphatidyl choline-bound Cu/Zn SOD (PC-SOD, 10(4), 10(5) U/kg) or a vehicle solution by intraperitoneal injection from postnatal 3-4 to postnatal 7-8 weeks of age. PC-SOD treatment attenuated progression of motor dysfunction, prevented denervation muscle atrophy, and delayed degeneration of spinal motoneurons in wobbler mice. This raises the possibility that PC-SOD may have therapeutic potential in human motoneuron disease. PMID- 7496171 TI - Blocking of NMDA receptors during a critical stage of development reduces the effects of nerve injury at birth on muscles and motoneurones. AB - Blocking of NMDA receptors during a critical stage of development reduces the effects of nerve injury at birth on muscles and motoneurones. Injury to the sciatic nerve at birth causes many motoneurones to soleus and extensor digitorum longus (EDL) muscles of rats to die. This is reflected in a reduction of motor units in these muscles. In the soleus only 4 (12.3%) motor units remain while 10 (24.3%) remain in the EDL, showing that soleus alpha motoneurones are more sensitive to nerve injury at birth. Treatment with MK-801, an NMDA receptor blocker, rescues a proportion of motor units in both muscles, so that in the soleus 11 (36%) and in the EDL 17 (42%) of motor units survive. This loss of motor units results in muscle weakness and a reduction in force of both muscles. Treatment with MK-801 reduces the effect of nerve injury, so that muscles of treated animals are stronger and weigh more. Cross-sectional area and muscle fibre number in EDL muscles were assessed and found to be dramatically reduced after nerve injury at birth, so that the area was 20% of control, with only 13% of fibres remaining. Moreover the majority of the remaining EDL muscle fibres which are normally fast are converted into slow type I fibres, with 68% of fibres expressing slow myosin compared with 3% in control EDL muscles. In animals treated with MK-801 only 47% of muscle fibres are lost after nerve injury at birth, hence the area of the muscle is greater (51% of control). The change of muscle phenotype induced by nerve injury is prevented and the muscle fibre composition resembles that of normal EDL muscles in that 4% of muscle fibres express slow myosin compared with 3.5% in control EDL muscles. Thus, blocking NMDA receptors with MK-801 shortly after nerve injury at birth reduces the loss of motor units and this is directly reflected in an improved performance of the affected muscles. PMID- 7496173 TI - A new mutation associated with MELAS is located in a mitochondrial DNA polypeptide-coding gene. AB - We report a patient with mitochondrial encephalomyopathy, lactic acidosis and stroke-like episodes (MELAS) who harbored a novel missense mutation at mtDNA position 9957 in the gene specifying subunit III of cytochrome c oxidase (COX III). This T-->C transition converted Phe-251, a highly conserved amino acid in the C-terminus of the polypeptide, to Leu. The mutation, which was not present in 107 normal controls or in 57 patients with various mitochondrial diseases, was heteroplasmic in both muscle and blood of the proband and in blood from his asymptomatic mother. These results provide evidence that the MELAS clinical phenotype can be due not only to mutations in mtDNA-encoded tRNA genes, but in polypeptide-coding genes as well. PMID- 7496174 TI - Chronic progressive external ophthalmoplegia with ragged-red fibers: clinical, morphological and genetic investigations in 43 patients. AB - The evaluation of the severity of progressive external ophthalmoplegia (PEO) with ragged-red fibers in muscle, at the onset of the disease, when PEO is most often the only presenting symptom, is a difficult problem in neurological practice. In order to address that issue, we have performed a comparative analysis of the clinical, morphological and molecular characteristics of 43 patients affected with that form of ocular myopathy. Quantification of mitochondrial accumulation was performed with an image analysis application on muscle sections stained with succinate dehydrogenase histochemical reaction. The proportion of muscle fibres appearing as cytochrome c oxidase deficient was used as an index of the muscle energy defect. Muscle mitochondrial DNA deletions were detected, localized and quantitated by Southern blot analysis. Point mutations were screened in five transfer RNA genes in the mtDNA (tRNA(Leucine (UUR)), tRNA(Lysine), tRNA(Glutamine), tRNA(Isoleucine) and tRNA(Formylmethionine)) by a denaturing gradient gel electrophoresis technique. This investigation confirmed the high frequency of mtDNA deletions or point mutations in PEO. At the onset of the disease, no clinical, morphological or molecular features could predict whether PEO would remain isolated or become part of a more severe multisystem disease. However, patients with mtDNA deletions were characterized by more severe ophthalmoplegia of earlier onset. Their muscle alterations were roughly parallel in severity to the proportion of deleted mtDNA molecules in muscle. Patients with a multitissular disease and mtDNA deletions were always sporadic cases and their clinical presentation was, most often, closely related to Kearns Sayre syndrome. PMID- 7496178 TI - [Future prospects of surgical gynecology in treatment of cervix carcinoma]. AB - Both the gain of new tumor biological insights, particularly the relevance of tumor oxygenation and vascularity in addition to tumor volume for disease progression and resistance towards therapy, and the development of novel ablative and reconstructive pelvic surgical techniques may lead to a paradigm shift in the treatment of cancer of the uterine cervix. With respect to stage, size, and predicted tumor biological features, either less radical or extended radical surgical treatment should be investigated in clinical trials with standard therapy (abdominal radical hysterectomy and pelvic/periaortic lymph node dissection for early-stage disease and percutaneous/intracavitary radiation for locally advanced disease) as control. Improving the therapeutic index and increasing the therapeutic window in cervical cancer appear to be an exciting challenge for surgical gynecology in the future. PMID- 7496175 TI - Isometric strength and thickness relationships in human quadriceps muscle. AB - The isometric Maximum Voluntary Contraction (MVC) of the right quadriceps femoris muscle was measured in 82 females and 58 males using a chair dynamometer and their mid-thigh quadriceps thickness was measured using real-time B-mode ultrasound. Statistically significant relationships (ranging from P < 0.001 to P < 0.0001) were found between quadriceps MVC, quadriceps thickness and body weight for both males and females. A significant inter-sex difference was present in the MVC-weight and MVC-thickness relationships, with males tending to have a higher MVC for a given body weight (P < 0.0001) and for a given muscle thickness (P < 0.0001). The inter-sex difference in the force-size relationship may relate to the previously demonstrated larger size of type 2b fibres and to the greater force generating capacity of these fibres compared with type 1 fibres. PMID- 7496176 TI - Generalized lysosomal storage in Yunis Varon syndrome. AB - Muscle biopsy in a neonate with features of Yunis Varon syndrome revealed a vacuolar myopathy with evidence of lysosomal storage disease. Similar vacuoles were also present in heart, cartilage, central nervous system and cultured fibroblasts. Although the histologic findings in the central nervous system resembled those of infantile acid maltase deficiency, the essayed lysosomal enzymes were normal. Chromatography of urine revealed abnormal bands of unidentified oligosaccharides. This is the first report of generalized storage disease in Yunis Varon syndrome. The biochemical defect is unknown. PMID- 7496177 TI - A severe muscular dystrophy patient with an internally deleted very short (110 kD) dystrophin: presence of the binding site for dystrophin-associated glycoprotein (DAG) may not be enough for physiological function of dystrophin. AB - We report a 4-yr and 5-month-old boy with severe clinical features of an early onset Duchenne muscular dystrophy, who had a very short (110 kDa) dystrophin at the sarcolemma. The patient had a large deletion (exons 2-44) of the dystrophin gene which was predicted to cause a reading frame shift. Sequence analysis of dystrophin mRNA in muscle revealed an alternatively spliced gene product from exons 1 to 51 that caused restoration of the reading frame, in addition to an mRNA corresponding to the DNA deletion. A consistent result was obtained by immunocytochemical analysis of muscle; i.e. positive staining for dystrophin at the sarcolemma using antibodies against the C-terminus, cysteine-rich region and last three of 24 repeat units of the central rod-domain, but not for the remaining antibodies for dystrophin that recognize the N-terminal and proximal rod-domains. Immunostaining for dystrophin-associated glycoproteins (DAGs: 43 and 50 K) and merosin were preserved. Utrophin staining was positive but fainter than other DMD muscles. These results suggest that an extremely short dystrophin lacking the entire actin-binding site in the N-terminus cannot function properly even if the protein possesses the putative DAG-binding cysteine-rich and the C terminal domains, and still has an ability to associate with sarcolemmal membrane. PMID- 7496179 TI - [The "Vienna School" of surgical gynecology as an important component of modern therapeutic concepts]. AB - During the last years, clinical applications and studies of gynecologic endoscopy have gained increasing importance. With regard to vaginal surgery, questions arise about single and combined use of endoscopy and vaginal techniques. Beside sufficient pretherapeutic assessment and patient information, evaluation of these indications requires prospective randomized and comparative trials. Thus, the clinical impact of combined endoscopic and vaginal techniques can be assessed on the basis of objective data. The alternative to this kind of evaluation of new surgical techniques is uncontrolled human experimentation which is neither scientific nor ethical. PMID- 7496180 TI - [The transvaginal surgical school in Austria. Retrospect-present-future ]. AB - The tradition of transvaginal hysterectomy started in 1878 and is still alive. A recent survey showed that two thirds of all hysterectomies performed in Austria are done transvaginally. Although indications for hysterectomy have changed quite fast during the last years, without doubt, hysterectomy remains an important alternative when conservative treatment fails. If hysterectomy is to be performed, the transvaginal route should be taken into consideration. Thus it is our task to pass our knowledge to those unfamiliar with the vaginal technique. PMID- 7496182 TI - [The gate the Kaiser opened for women. Rotenhausgasse (1), Vienna-Alservorstadt]. AB - This paper refers to one of the entrances to the former 'delivery house' of the old Vienna General Hospital, namely the door in Rotenhausgasse. It was used for 70 years, from 1784 to 1854. According to the wishes of Emperor Joseph II, unmarried women could be supported by specialists, anonymously and thus escaping shame. Connected with it was the protection of the newborn against exposure and infanticide. PMID- 7496181 TI - [The future of surgical gynecology: from dogma to science--from science to practice standard]. AB - In the future of gynecologic surgery, there will be a reduction of traditional and an increase of endoscopic surgery. Development, progress, and realization of new technologies have to be based on scientific results, as it has been shown for the surgery of early-stage ovarian cancer. Endoscopy is already part of daily routine surgery in nearly all gynecologic departments, although important questions like the optimal technique, proper indications, and the consequences of these new technologies regarding the course of ovarian cancer are still not answered. Especially gynecological surgery has to turn away from a dogmatic way of thinking to a critical science-oriented practice in the future. PMID- 7496183 TI - [Age and sex specific differences in bone density of 4,000-year-old individuals. Bone density measurement of early Bronze Age femurs from excavations in Unterhautzenthal, Lower Austria]. AB - Bone density measurements of 14 individuals (early Bronze Age, 2200-1600 BC), analyzed by dual-energy absorptiometry, showed a distinct difference between women and men. In men, there was a high bone density (+17.9%-0.290 g/cm2 difference). In females, a constant decrease in bone density was found after the age of 20-25 years (from 1.2 g/cm2 at 20 years to 0.8 g/cm2 at 40 years of age). With caution, the results are comparable with those of the present time. PMID- 7496184 TI - [Common history and prospects in surgical gynecology of the Charite and the Vienna University Clinic]. AB - In a review, the historical connections between the medical centers of the University of Berlin (Charite) and the University of Vienna are described. The present status is reported and several suggestions for a further development are given. PMID- 7496185 TI - [Effect of needle suspension on continence mechanisms]. AB - OBJECTIVE: To compare the separation in time of urethral and bladder pressure pulses and urodynamic parameters after successful and unsuccessful anterior colporrhaphy and needle suspension of the bladder neck in patients with stress urinary incontinence. STUDY DESIGN: Retrospective evaluation of urethral pressure profiles at rest and under stress in 10 successfully and 10 unsuccessfully operated patients. RESULTS: In the successfully treated group, the onset of the urethral pressure increase preceded that of the bladder pressure by an average of 300 ms. Pressure transmission ratios in the 3 proximal quartiles of the urethra were significantly improved after successful surgery. Women who remained incontinent showed neither an advanced onset of the urethral pressure pulse during stress nor improved pressure transmission ratios. CONCLUSION: Successful anterior repair and needle suspension of the bladder neck support the bladder neck and thus improve the pressure transmission to the urethra. Improved pressure transmission is associated with the finding that pressure increases in the urethra precede those in the bladder in continent women. PMID- 7496186 TI - [Urethrovesical morphology in continent women in spinal anesthesia]. AB - OBJECTIVE: To assess the influence of spinal anesthesia on bladder neck position and a clinical stress test in continent women. METHODS: In a prospective investigation, 14 women underwent urodynamic, sonographic and clinical assessment during spinal anesthesia. Results were compared to those obtained immediately preoperatively in the same patient. RESULTS: During spinal anesthesia, the bladder neck was found to be located significantly lower and more posterior, and in 4/7 parous patients (0/7 nullipara) the clinical stress test was positive. CONCLUSION: These data provide additional evidence for the importance of neuromuscular function in the etiology of pelvic floor dysfunction and genuine stress incontinence. PMID- 7496187 TI - [5 years results of incontinence operations]. AB - OBJECTIVE: To evaluate results 5 years after anterior colporrhaphy with or without needle suspension of the bladder neck and after Burch colposuspension. METHODS: 186/291 women (64%) operated on between 1989 and 1990 underwent clinical and urodynamic reevaluation. RESULTS: Objective continence rates after anterior colporrhaphy were 80% (20/25), 46% (22/47) and 12.5% (1/8) in patients with first , second- and third-degree stress incontinence according to Ingelman-Sundberg. Continence rates in patients with second- and third-degree stress incontinence were 59% (29/49) after additional needle suspension and 86% (50/58) after the Burch procedure. CONCLUSIONS: Anterior colporrhaphy can cure mild stress incontinence but is not indicated for severe incontinence. The Burch procedure provides the best continence rates in patients with severe stress incontinence. PMID- 7496188 TI - [Horizon 2000: what are the perspectives for African trypanosomiasis?]. PMID- 7496190 TI - [Human African trypanosomiasis. The current epidemiologic situation, an alarming recrudescence of the disease]. PMID- 7496189 TI - [The role of environmental factors in the current recrudescence of human African trypanosomiasis]. AB - The authors highlight the major target for the prevention of human african trypanosomiasis (HAT) i.e. decrease of parasites reservoir. For this purpose, they state that it is necessary to detect HAT patients, to treat them, and to break down the contact between human and vectors. Then, they review the environmental factors that could be involved in the HAT outbreak. A political direction would strengthen the HAT prevention by the increase of resources attributable to the disease. But, choice of priorities is based on technical criteria which are not in favour of HAT. Many difficult situations, occurring at the borders of concerned states, are restricting factors for a global and co ordinated prevention. Moreover, internal political disturbances and local conflicts lead to a mismanagement of health care departments. These conflicts lead also to a migratory flow of populations; thereafter, the individuals have no access to medical structure. Beside the economical and political factors, some others have a great importance: human behavior, estimation of risk by the authorities, psychological perception of the disease by the population. The part of these different factors must be perfectly identified to avoid any disturbance in the actions of HAT prevention. The least failure leads to a quick increase of parasites transmission, and the HAT control has to start all over again. PMID- 7496191 TI - [Acute Trypanosoma brucei gambiense meningoencephalitis detected by papillary edema]. AB - A 26 year old woman, living in Brazzaville, was referred by her ophthalmologist about papillo-oedema. Neurological examination showed frontal syndrome and papillo-oedema without another sign of intracranial hypertension. Electroencephalogram revealed bilateral delta waves and bifrontal points. Significant inflammatory syndrome was noted. Cerebrospinal fluid examination showed trypanosomes with an elevated rate of protein and cells. Treatment by tetracosactide and eflornithine gave a good recovery. Authors are stressing the disconcerting feature of this clinical form. An urban from of the disease and the fight against african trypanosomiasis are also discussed. PMID- 7496192 TI - Identification of trypanosomes in animals, humans and Glossina. AB - A number of biochemical methods are now available for the identification of African trypanosomes. The method of choice depends on the number of trypanosomes present in the sample and the taxonomic level required. DNA probes based on repetitive DNA elements allow identification to subgeneric (e.g. Trypanozoon), species (e.g. Trypanosoma congolense, T. simiae) or subspecific (e.g. T. congolense savannah) levels. These probes are particularly useful for identification of trypanosomes in the fly midgut, where sufficient numbers are present to allow simple dot blot hybridization to be used (> 100). Greater sensitivity has been achieved by amplification of these repetitive DNA sequences by PCR (polymerase chain reaction), so enabling the small numbers of trypanosomes found in the fly mouthparts to be identified (> 1). At the subspecific level, isoenzyme analysis and latterly RFLP (restriction fragment length polymorphism) analysis have been widely used to characterize isolates within the T. brucei species. Two other techniques, karyotype analysis and RAPD analysis, are also useful for fingerprinting isolates. Molecular karyotypes are produced by size fractionation of chromosomal DNAs by PFGE (pulsed field gel electrophoresis). RAPD (random amplified polymorphic DNA) is a PCR-based technique, using arbitrary primers to generate a fingerprint consisting of 20 or so bands. PMID- 7496193 TI - A new approach for the pathogenesis of human African trypanosomiasis. AB - From Trypanosoma brucei brucei, a molecule has been isolated which triggers the production of IFN-gamma by CD8+ T-cells. IFN-gamma modulates events in the hosts immune- and nervous system and provides a growth stimulus for the parasites. Furthermore, a molecule with IFN-gamma-like immunoreactivity has been detected in rat dorsal root ganglion cells and certain neurons in the brain. This neuron derived IFN-gamma-like molecule differs in molecular weight from lymphocyte derived IFN-gamma but shares important biological activities with the latter, which includes a growth stimulus for trypanosomes. Trypanosomes localized to sensory ganglia and infected rats develop a severe thermal hyperalgesia. Intrathecal injection of IFN-gamma causes in rats a sustained phase of nociceptive flexor reflex facilitation, which can be partially blocked by nitro-L arginin-ester, an inhibitor of nitric oxide synthase, indicating that nociceptive effects of the IFN-gamma is mediated by activation of the L-arginin-nitric oxide pathway. PMID- 7496195 TI - [Defense mechanisms in trypanosomiasis]. AB - Quantitative and functional alterations in macrophages are observed in trypanosomiasis. Two molecules from macrophages exert potent anti-microbial effect: Nitric Oxide (NO) and Tumor Necrosis Factor-alpha (TNF-alpha). The role of NO in trypanosomiasis is investigated at first on a murine parasite. Trypanosoma musculi, at first and then on Trypanosoma brucei gambiense and T. brucei brucei. Macrophages from T. musculi-infected mice synthesize NO and their trypanostatic activity is correlated with NO production. In vitro activation of macrophage NO synthase by IFN-gamma induces a trypanostatic activity and TNF alpha is involved in NO synthase induction. High serum levels of TNF-alpha are correlated with disease severity in human African trypanosomiasis. TNF-alpha is increased in supernatants of leucocyte and trypanosome cocultures. TNF-alpha exerts a strong anti-trypanosomal effect. Messengers RNA of TNF-alpha are detected in monocytes after 8 hours of coculture with trypanosomes. Macrophage effector molecules participate with other immune effector mechanisms in resistance of host to trypanosomes. PMID- 7496194 TI - Trypanosoma brucei products activate components of the reactive response in astrocytes in vitro. AB - A study was made of the effects of T. b. brucei and the disrupted parasite material on different components of the reactive response of astrocytes in primary cultures prepared from neonatal rats and C6 glioma cells. The effects were compared with lipopolysaccharide (LPS) and fragments of the C6 cells. The disrupted trypanosome material, LPS and C6 fragments caused dose--and time- dependent alterations in morphology of the primary cultures from flat to stellate shape, increases in levels of glial fibrillary acidic protein (GFAp) and enhanced MHC class I and II expression. Exposure to trypanosome material and C6 fragments caused marked increases in phagolysosomes and lysosomes in the primary cultures. The findings demonstrate that T. b. brucei products and astrocyte cell debris are internalized and initiate astrogliosis in vitro. PMID- 7496197 TI - [Current therapy of trypanosomiasis]. AB - The authors review the available products used for human african trypanosomiasis (HAT) chemotherapy: pentamidine, suramin, melarsoprol and the new compound DFMO. The administration of pentamidine at the beginning of the nervous stage, when the number of cells in the cerebrospinal fluid do not exceed 20/mn3 is a new approach for HAT treatment. At this time of the disease, patients generally are healthy and the pentamidine therapy avoids the use of the toxic melarsoprol (Arsobal). An alternative protocol for Arsobal therapy (2, 16 mg/kg/d for 10 consecutive days) has been described from pharmacokinetics data to decrease the rate of relapses and the duration of hospital care. Efficacy and tolerance of this new protocol must be evaluated by randomised clinical trials. Preliminary data of clinical trials using short-term DFMO therapy are encouraging. DFMO therapy be less expensive. From its efficacy and tolerance, DFMO is a choice chemotherapy for HAT treatment, especially in the case of resistance to usual trypanocides. Both MLD 73811 and IMOL 881 are new trypanocidal compounds, effective on Trypanosoma brucei rhodesiense and T. b. gambiense. In addition, IMOL 881 is effective on the animal trypanosomes, T. evansi and T. equiperdum. Waiting for the availability of these new products, classical trypanocides remain the basis of HAT treatment. PMID- 7496196 TI - [The detection of anti-galactocerebroside autoantibodies in human African trypanosomiasis]. AB - The pathogenesis of the central nervous system (CNS) damage in human african trypanosomiasis (HAT) is unknown. In view of an immunological mechanism, as in another trypanosomiasis, Chagas' disease, the causative agent of which is Trypanosoma cruzi, we have searched autoantibodies directed against glycosphingolipids of CNS. Detection and characterization of autoantibodies were performed by ELISA and detection after thin-layer chromatography of glycolipids with sera of an experimental model of HAT in sheep and sera of patients suffering of HAT from Cote d'Ivoire and Congo. The predominant reactivity of these sera, was characterized with galactocerebrosides, the major glycolipids of the myelin. Autoantibodies were detected in 42.8% and 25% of patients' sera, respectively from Cote d'Ivoire and Congo. The proportion of these antibodies increased dramatically to 72% in sera of patients with neurological symptoms. Anti galactocerebroside antibodies were also found in CSF of 24.4% of Congolense patients. The pathogenic significance of these anti-galactocerebroside antibodies remains to be determined. They may constitute a predicative marker for the neurological improvement in HAT. PMID- 7496198 TI - [2 epidemics of arsenical encephalopathy in the treatment of trypanosomiasis, Uganda, 1992-1993]. AB - Since 1988, the french non-governmental organisation Medecins Sans Frontieres is running a control program of human african trypanosomiasis in the district of Moyo, North-Uganda. Between 1988 and 1993, more than 7,000 patients were diagnosed and treated. Since 1988, it has been noted that incidence of melarsoprol reaction had increased systematically between June and October of each year, indicating strong seasonal variation. In 1992 and 1993, two outbreaks of arsenical reactive encephalopathy (ARE) occurred in the sleeping sickness center of Adjumani. The incidence of ARE suddenly exceeded 10% of the patients treated by melarsoprol during August 1992 and September 1993. The onset of 80% of those "epidemic" cases, occurred between the 5th and the 11th day of treatment. Two retrospective studies were conducted in 1992 (75 cases) and in 1993 (51 cases). Among the risk factors studied, two appear to increase the risk of ARE: the prescription of thiabendazole to treat strongyloidiasis during the melarsoprol cure and the bad general clinical conditions of patients. These observations suggest that exogenous co-factors could be involved in the occurrence of ARE. Recommendations were to avoid administration of diffusible anti-helminthic treatment during the cure, and to improve the general conditions of patients before the cure of melarsoprol. PMID- 7496199 TI - [An efficacy trial on Trypanosoma brucei brucei of molecules permeating the blood brain barrier and of megazol]. AB - Human African trypanosomiasis (HAT) is a major public health problem in 36 sub Saharan African countries and around 50 million people are classed as "at risk". About 25,000 new cases of the disease are reported annually by the World Health Organisation (WHO). This disease is fatal if untreated. As for now, chemotherapy is unsatisfactory and relies on a few drugs which show two major problems. The first is pharmacokinetics involving the passage through the blood-brain barrier. The second concerns toxicity and adverse side-effects of drugs used to treat this disease. New trypanocides should be safe, effective without toxicity. This study reports the action of 45 drugs, known to pass through the blood-brain barrier and belonging to different therapeutic classes, and also the megazol, a nitrothiadiazole derivative, on Trypanosoma brucei brucei AnTat 1-9 in vitro in acellular semi-defined medium. Results showed that some drugs did not modify the parasitic growth, and others were either trypanostatic or trypanocide. These last drugs were tested in vivo on T. b. brucei An-Tat 1-9 infected Swiss mice. Only megazol was shown to be effective and trypanocide. This compound might trigger the production of oxygen derivatives and free radicals-which have toxic effects on the trypanosome metabolism. PMID- 7496200 TI - [Rational concepts and the study of active molecules against various trypanosomiases]. AB - Several sets of compounds, active against different trypanosomes, Trypanosoma brucei and Trypanosoma cruzi are presented, the lethal doses for some of them being less than the micro-molar concentration. These compounds are designed by taking advantage of two metabolic features of these parasites, glucose metabolism and oxidative stress. PMID- 7496202 TI - [The sleep-wake cycle during Trypanosoma brucei rhodesiense human African trypanosomiasis in 2 French parachutists]. AB - Polysomnography was performed in two military personnel who contracted human African trypanosomiasis (HAT) in Rwanda. They were evacuated to France and gramined during the fourth, sixth and eleventh months of evolution. Patient A presented a meningoencephalitis with Trypanosoma brucei rhodesiense (T. b. r.) in the cerebrospinal fluid. Between the second and third treatments with melarsoprol, the patient presented an acute encephalitic attack with pyramidal and cerebellar symptoms, which were improved by corticotherapy and disappeared progressively in 6 months. Patient B presented a major inflammatory syndrome with T. b. r. in the blood and the medulla. Cerebrospinal fluid and serology remained normal. The patient received only one session of melarsoprol treatment. Polysomnography recordings were continuously taken on an ambulatory Oxford Medilog system during 48 hours. The different stages of vigilance were scored according to the classical Rechtschaffen and Kales criteria. During the fourth month, both subjects presented several long diurnal naps, with the presence of rapid-eye-movement sleep at the end of the sleep episodes. These abnormalities disappeared progressively during the following recordings. Electroencephalographic abnormalities were observed in patient A, particularly during the eleventh month. They consisted in transitory arousal phases concomitant with myoclonic jerks and with the persistence of slow waves during slow-wave sleep. The various anomalies noted in the two patients allow us to recommend the supervision of the evolution of HAT by polysomnographic recording in a sleep laboratory. PMID- 7496201 TI - [The distribution of sleep and wakefulness in human African trypanosomiasis]. AB - Last century, patients with human African trypanosomiasis were described as sleepy by day and restless by night, and physicians referred to this condition as sleeping sickness. Such a description could have evoked a disturbance of circadian rhythms. However, it is only in 1989 that the first 24-hour recording was performed by our team in Niamey (Niger) in a patient with sleeping sickness. The patient was a Niger-born farm worker who had contracted the disease near Gagnoa (Cote d'Ivoire). Polysomnographic recordings (electroencephalogram, EEG, electrooculogram, electromyogram, electrocardiogram, buccal and nasal airflow, and chest respiratory movements) showed a disappearance of the circadian distribution of sleep and wakefulness, which tended to occur evenly throughout day and night, with a sleep-wake alternation of approximately 80 minutes. Two investigations were conducted thereafter. The first one was done at Daloa (Cote d'Ivoire) in 8 patients who were recorded during two 24-hour periods, with and without hourly blood samples; the second at Brazzaville (Congo) in 10 patients recorded for 24 hours before and after treatment with melarsoprol. All patients were at the stage of early meningoencephalitis. At Daloa, polysomnographic recordings were taken on two 8-channel EEG machines (Alvar Minihuit, and T3 ECEM), as well as on a portable Oxford Medilog 9000 system from the same electrodes. Sleep and wake structure was altered in the most severely sick patient, the EEG trace being loaded with slow waves. Stages 1 and 2, and stages 3 and 4 could not be distinguished from one another. In the other patients, all sleep stages were easily scored. No difference was seen between recordings, regarding blood collection.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7496203 TI - Sleep and timekeeping changes, and dysregulation of the biological clock in experimental trypanosomiasis. AB - The rest-activity and body temperature 24 hours cycles, as well as the pattern of spontaneous sleep, were investigated in rats after infection with Trypanosoma brucei brucei. In the infected rats, which were entrained to a 12 hours/12 hours photoperiod, a considerable hypokinesia was detected during the hours of darkness. In most of the infected animals, the body temperature cycle displayed a lower amplitude and an advance of about 3 hours in respect to control rats; in addition, the body temperature rhythm was not significant in some infected rats. The relative proportion of slow wave synchronized sleep, as well as the rapid-eye movement (REM) latency, were significantly reduced in the infected animals, in which sleep was considerably fragmented. The induction of Fos (the protein encoded by the immediate early gene c-fos), in response to light stimulation during the early subjective night, was severely impaired in the hypothalamic suprachiasmatic nuclei in trypanosome-infected rats. Altogether these data point out a disruption of locomotor activity and body temperature 24 hours cycle and a major disorganization of sleep during experimental trypanosomiasis. In addition, our findings indicate that the molecular and functional correlates of the synchronizing action of the suprachiasmatic nuclei, which play a major role of biological clock of endogenous biological rhythms, could be altered during trypanosome infection. PMID- 7496204 TI - Disruptions in the secretion of cortisol, prolactin, and certain cytokines in human African trypanosomiasis patients. AB - It has been shown previously that sleeping sickness at the stage of meningoencephalitis manifests itself as a significant disturbance in the circadian rhythm of sleep-wakefulness. The objective of the current study was to examine the extent of circadian disruption in infected patients by measuring 24 hours patterns of plasma cortisol, an example of a classical circadian rhythm relatively independent of sleep, and prolactin, a primarily sleep-related rhythm. Plasma levels of certain cytokines were also measured to examine the immunopathogenesis of human African trypanosomiasis. An attempt was made to relate any circadian disruptions to the severity of the disease. The three most advanced patients demonstrated circadian disruptions in cortisol, prolactin and sleep-wake rhythms. The prime cytokine factor that correlated with the progression of the disease in humans was interferon-gamma, levels being 7- to 12 fold higher in the patients without any circadian rhythms. Our findings support the hypothesis that human African trypanosomiasis induces selective changes in the suprachiasmatic nucleus, important as a pacemaker for biological rhythms, resulting in disruptions of circadian rhythmicity in advanced stages of the disease. PMID- 7496206 TI - [Maintenance of the relation between the pulsed secretion of hormones and the internal sleep structure in human African trypanosomiasis]. AB - In order to determine whether sleep disturbances would affect the hormonal patterns and the normal relationships between hormone pulses and sleep stages, the 24-hour profiles of cortisol, prolactin and plasma renin activity (PRA) were analysed in 6 sleeping sickness patients studied at Brazzaville and in 5 healthy African controls studied in Abidjan. Polysomnographic recordings were done continuously and blood was taken every 10 minutes throughout the 24-hour period. Plasma was analyzed for cortisol, prolactin and PRA. The circadian rhythm of cortisol, considered as an example of an endogenous rhythm was attenuated in all the patients but one, but as in normal subjects, slow wave sleep (SWS) remained associated with the declining phases of the secretory episodes. Prolactin and PRA profiles, which are strongly influenced by the sleep-wake cycle did not show the increase normally associated with long sleep periods and reflected the spreading of sleep and wakefulness throughout the 24-hour period. However, rapid-eye movement (REM) sleep began in sleeping sickness patients, as in normal subjects, during the descending phases of prolactin pulses. In both groups, PRA reflected the sleep stage distribution with non rapid-eye movement (NREM) sleep occurring during the ascending phases and REM sleep during the descending phases of the oscillations. However, in sleeping sickness patients, the marked sleep fragmentation often did not allow sufficient time for PRA to increase significantly, as observed with regular NREM-REM sleep cycles. These results demonstrate that, together with the disruption of the sleep-wake cycle, there are profound differences in the temporal organization of the 24 hour hormone profiles in human African trypanosomiasis.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7496205 TI - [The nyctohemeral rhythm of melatonin is preserved in human African trypanosomiasis]. AB - We studied plasma melatonin profiles by radioimmunoassay in nine patients suffering from human african trypanosomiasis and six healthy controls matched according to the age and the photoperiodic conditions. The circadian periodicity of the sleep-wake cycle was disturbed proportionally to the degree of severity of the disease. On the contrary, the patients' plasma melatonin profile was similar to the controls' one. These results suggest that, beside the master clock generating the main circadian rhythms (sleep-wake, melatonin and core temperature rhythms), an additional regulating system of the melatonin rhythm could be involved. PMID- 7496207 TI - [Trypanosomiases, diseases of the future: their perspectives and their uncertanties]. PMID- 7496208 TI - [The experience of the first 100 surgical laparoscopies]. PMID- 7496209 TI - [Problems of the therapeutic approach in subperitoneal wounds to the rectum]. AB - We present four observations about the treatment of subperitoneal rectal injuries. Left iliac colostomy and cystostomy are usually necessary as first time operations in those cases with associated urogenital injuries. For the subperitoneal rectal injuries simple suture of rectum and drainage are sufficient. PMID- 7496210 TI - [Clinical and cytogenetic observations on carcinoid tumors]. PMID- 7496211 TI - [Secondary hyperparathyroidism]. AB - Secondary (renal) hyperparathyroidism appears in chronic renal failure, sometimes in patients on chronic dialysis. Other causes includes rickets and osteomalacia. These diseases are associated with poor calcium and vitamin D absorbtion from the small bowel. Two patients with chronic renal failure maintained on chronic haemodialysis from two and three years, respectively underwent subtotal parathyroidectomy: removal of three glands and preserving a half of a gland in situ. The diagnosis and surgical indication was made upon clinical (bone pain and severe itching), radiological (demineralisation, ectopic calcifications) and biochemical (hypercalcemia, hyperphosphoremia, increased values of alkaline phosphatases) arguments. Postoperatively the improvement is defined by a return to normal in the clinical, laboratory and radiological parametres. The most appropriate operation for secondary hyperparathyroidism is still unresolved one of two techniques is performed according to the preference of the surgeon: subtotal parathyroidectomy or total parathyroidectomy with autotransplantation of parathyroid fragments into forearm muscle. PMID- 7496212 TI - [Stenosing hypertrophic antromyopathy]. AB - The authors describe a case of insufficiency of gastric evacuation, due to the a very rare injury of the antrum which they called:--"hypertrophic and stenosing antromiopathiae". By similarity with the pilor hypertrophy, but without altering the pilor which is intact, the hypertrophy involves especially the gastric antrum, the muscular tunic--mainly the circular fibres in the absence of any acute or chronic inflammatory process. In the interstice between the muscular fibres, there appear collagen fibres in spiral disposition, probably in a contraction. Due to the similitude of the clinical syndrome, the described disease could be ranged among the stenosant diseases of the pyloric pole of the stomach. Out of these, more frequently pointed out, although very rare too, seems to be the pilor hypertrophy at adults. Mention must be made that the two diseases are completely different, because in the "Hypertrophic and stenosing antromiopathiae", the pilor is macroscopic and at the same time normal from the histopathologic aspect point of view. PMID- 7496213 TI - [Cystic lymphangioma of the adrenal gland]. PMID- 7496214 TI - [Odelberg's lipomatosis]. PMID- 7496215 TI - [A report of 6 cases of traumatic ruptures of the duodenum]. PMID- 7496216 TI - Polyunsaturated fat and the risk of cancer. PMID- 7496217 TI - Publishing information about patients. PMID- 7496218 TI - Atopic eczema. PMID- 7496219 TI - New arrangements for specialist training in Britain. PMID- 7496220 TI - Evaluating new surgical procedures. PMID- 7496222 TI - GMC finds doctors not guilty in consent case. PMID- 7496221 TI - Corticosteroids in the management of croup. PMID- 7496223 TI - WHO sets new guidelines for air pollution. PMID- 7496224 TI - Black and Asian nurses in the NHS report harassment. PMID- 7496225 TI - French protest at health budget cuts. PMID- 7496226 TI - Doctor wins claim over patent ownership. PMID- 7496227 TI - Foreign doctors strike over new French law. PMID- 7496228 TI - Minister defends immunisation campaign. PMID- 7496229 TI - Health network will work to reduce poverty. PMID- 7496230 TI - Australian wins right to fight tobacco company. PMID- 7496231 TI - Litigation Authority to focus on malpractice. PMID- 7496232 TI - Fatty acid proportions in cholesterol esters and risk of premature death from cancer in middle aged French men. AB - OBJECTIVE: To assess the association of proportions of fatty acids in cholesterol esters with the risk of premature death from cancer in middle aged men. DESIGN: Prospective cohort study. SETTING: Paris, France. SUBJECTS: 3277 working men aged 36-52 in 1981-5. MAIN OUTCOME MEASURES: Cancer mortality during an average of 9.3 years of follow up. RESULTS: 59 men died of cancer during follow up. The age adjusted relative risks for men in the highest thirds of the distribution of the proportions of linoleic, palmitoleic, and oleic acid in cholesterol esters as compared with those in the corresponding lowest thirds were 0.16 (95% confidence interval 0.05 to 0.51), 3.39 (1.63 to 7.05), and 4.22 (1.95 to 9.12), respectively. Adjustment for and stratification by smoking, alcohol consumption, serum cholesterol concentration, and body mass index did not alter the results. At the time of examination subjects with cancer had a lower intake of polyunsaturated fats, assessed by 24 hour recall, than those without cancer (13.2 v 17.4 g/day, P < 0.01). CONCLUSIONS: Monounsaturated and polyunsaturated fatty acids of cholesterol esters are strong biological markers that predict premature death from cancer in French men. Consistently, intake of polyunsaturated fats did not seem to increase the risk of death from cancer. The association of biological markers of dietary fat intake with incidence of and mortality from cancer should be investigated prospectively in other populations. PMID- 7496233 TI - Changes in population cholesterol concentrations and other cardiovascular risk factor levels after five years of the non-communicable disease intervention programme in Mauritius. Mauritius Non-communicable Disease Study Group. AB - OBJECTIVE: To study changes in the prevalence of risk factors for cardiovascular disease after a five year population-wide intervention programme promoting a healthy lifestyle in a developing country. DESIGN: Cross sectional cluster surveys in 1987 and 1992. Methodology included a two hour 75 g oral glucose tolerance test, measurement of body mass index, waist:hip ratio, basal lipid concentrations, and blood pressure; and a lifestyle questionnaire. SETTING: Mauritius, in the Indian Ocean. SUBJECTS: All adults aged 25-74 years residing in geographically defined clusters. MAIN OUTCOME MEASURES: Age standardised prevalence of categorical disease and risk factor conditions and mean levels and frequency distributions of continuous variables. RESULTS: Response rates were 86.2% (5080/5892) in 1987 and 89.5% (5162/5770) in 1992. Significant decreases were found in the prevalence of hypertension (15.0% to 12.1% in men and 12.4% to 10.9% in women); cigarette smoking (58.2% to 47.2% and 6.9% to 3.7% respectively); and heavy alcohol consumption (38.2% to 14.4% and 2.6% to 0.6% respectively). Moderate leisure physical activity increased from 16.9% to 22.1% in men and from 1.3% to 2.7% in women. Mean population serum total cholesterol concentration fell appreciably from 5.5 mmol/l to 4.7 mmol/l (P < 0.001). The prevalence of overweight or obesity increased, and the rates of glucose intolerance changed little. The population frequency distributions of blood pressure, serum lipid concentration, and a composite risk factor score shifted advantageously. CONCLUSIONS: Lifestyle intervention projects can be implemented and have positive effects in developing countries. A pronounced improvement in the population lipid profile in Mauritius was probably related to a change in the saturated fat content of a widely used cooking oil. PMID- 7496235 TI - Health effects of anticipation of job change and non-employment: longitudinal data from the Whitehall II study. AB - OBJECTIVE: To assess the effect of anticipating job change or non-employment on self reported health status in a group of middle aged male and female white collar civil servants. DESIGN: Longitudinal cohort study (Whitehall II study). Questionnaire data on self reported health status and health behaviour were obtained at initial screening and four years later, during the period when employees of the department facing privatisation were anticipating job change or job loss. SETTING: London based office staff in 20 civil service departments. SUBJECTS: 666 members of one department threatened with early privatisation were compared with members of the 19 other departments. MAIN OUTCOME MEASURES: Self reported health status measures and health related behaviours, before and during anticipation of privatisation. RESULTS: In comparison to the remainder of the cohort, the profile of health related behaviours of cohort members who faced privatisation was more favourable, both before and during anticipation of privatisation. There were no significant differences in the changes in health behaviours between cohort members moving into a period of job insecurity and the remainder of the cohort. Self reported health status, however, tended to deteriorate among employees anticipating privatisation when compared with that of the rest of the cohort. CONCLUSIONS: The application of a longitudinal design, allowing the same individuals to be followed from job security into anticipation, provides more robust evidence than has previously been available that anticipation of job loss affects health even before employment status has changed. PMID- 7496236 TI - Bed sharing and the sudden infant death syndrome. AB - OBJECTIVE: To determine whether infants who died of the sudden infant death syndrome routinely shared their parents' bed more commonly than control infants. DESIGN: Case-control study. SETTING: Southern California. SUBJECTS: 200 white, African-American, Latin American, and Asian infants who died and 200 living controls, matched by birth hospital, date of birth, sex, and race. MAIN OUTCOME MEASURES: Routine bedding (for example, crib, cradle), day and night time sleeping arrangement (for example, alone or sharing a bed); for cases only, sleeping arrangement at death. Differences in bed sharing practices among races. RESULTS: Of the infants who died of the syndrome, 45 (22.4%) were sharing a bed. Daytime bed sharing was more common in African-American (P < 0.001) and Latin American families (P < 0.001) than in white families. The overall adjusted odds ratio for the syndrome and routine bed sharing in the daytime was 1.38 (95% confidence interval 0.59 to 3.22) and for night was 1.21 (0.59 to 2.48). These odds ratios were adjusted for routine sleep position, passive smoking, breast feeding, intercom use, infant birth weight, medical conditions at birth, and maternal age and education. There was no interaction between bed sharing and passive smoking or alcohol use by either parent. CONCLUSIONS: Although there was a significant difference between bed sharing among African-American and Latin American parents compared with white parents, there was no significant relation between routine bed sharing and the sudden infant death syndrome. PMID- 7496234 TI - Individual variation in plasma cholesterol response to dietary saturated fat. AB - OBJECTIVE: To determine the extent to which plasma lipid concentrations of individuals are consistently sensitive to changes in saturated fats; to examine whether groups that consistently have large or small responses can be defined; and to identify factors which predict response of lipids to dietary change. DESIGN: A double crossover design in which two diets (S, providing 21% energy from saturated fat, and P, providing 10%) were followed for periods of six weeks in the sequence SPSP or PSPS. SETTING: 67 free living subjects, total cholesterol 5.5-7.9 mmol/l. MAIN OUTCOME MEASURES: Relation of cholesterol responses to repeated dietary changes and of potential predictors and cholesterol response. RESULTS: Similar average changes in cholesterol mask a wide range of individual responses. Response was not related to compliance. In all participants the change in cholesterol observed when the nature of dietary fat was changed on the two crossovers was correlated (r = 0.31, P = 0.01); the degree of correlation between the two sets of responses was greater in the 46 consistent responders than in the 21 variable responders (r = 0.71 v r = 0.21). Mean differences in cholesterol between diet S and diet P during the two crossovers were 1.16 (SD 0.35) mmol/l and 0.95 (0.26) mmol/l for consistent hyperresponders and 0.18 (0.26) mmol/l and 0.18 (0.25) mmol/l for consistent minimal responders. In consistent responders, changes in total cholesterol in response to increasing saturated fats correlated with baseline cholesteryl ester transfer activity (r = 0.32, P = 0.03); total cholesterol (r = 0.37, P = 0.01); triglycerides (r = 0.30, P = 0.04); and apolipoprotein B (r = 0.54, P = 0.01). CONCLUSIONS: There is a degree of consistency in cholesterol response to instructions to change dietary fat which is not explained by dietary compliance, and there are groups of consistent hyperresponders and minimal responders within a population of hypercholesterolaemic individuals. Several factors predicting response have been identified. These results have relevance to dietary approaches aimed at reducing the lipoprotein mediated risk of coronary heart disease. PMID- 7496237 TI - The incidence of malaria after splenectomy in Papua New Guinea. PMID- 7496238 TI - Twelve month outcome of depression in general practice: does detection or disclosure make a difference? AB - OBJECTIVES: To assess the extent to which the outcome of depression among primary care attenders may be affected by medical diagnosis or by feedback of questionnaire results in unrecognised cases. DESIGN: Prospective 12 month study including a randomised controlled trial of the effects of disclosure, with data on depression status and clinical management collected by questionnaire and interview. SETTING: Two group practices in north Liverpool. SUBJECTS: 1099/1444 (76%) consecutive adult attenders completed the Beck depression inventory, of whom 179 with scores of at least 14 were followed up. INTERVENTIONS: Disclosure of a random 45% (52/116) of depression scores to general practitioners for subjects whose depression was undetected. MAIN OUTCOME MEASURES: Depression status estimated by depression score at start of study and at six and 12 months, with subsample validation against ICD-10 criteria. RESULTS: Questionnaire response rates were 76% (136/179) at six months and 68% (122/179) at 12 months and were higher for women than men. The median depression score was 19 (interquartile range 15 to 22) initially, decreasing to 16 (11 to 23) at 12 months. The median depression score decreased significantly (two sided test, P = 0.019) in subjects whose depression was unrecognised at the index consultation but increased in those whose depression had been detected by their general practitioners. Disclosure of cases of unrecognised depression to general practitioners had no effect on outcome. Intention to treat was associated with a worse prognosis, although only a minority of subjects received adequate treatment. CONCLUSIONS: Disclosure of undetected depression did not improve prognosis. A diagnosis of depression in general practice should be considered simply as a marker of its severity. PMID- 7496239 TI - Fungal nail disease: a guide to good practice (report of a Working Group of the British Society for Medical Mycology). PMID- 7496240 TI - Income distribution and life expectancy: a critical appraisal. AB - In a series of papers published during the past decade Richard Wilkinson has advanced the view that income inequality is the key determinant of variations in average life expectancy at birth among developed countries. Yet a careful examination of the two sources of data on income distribution most often used by Wilkinson suggests that if they are analysed more appropriately they do not lend support to his claims. More recent data on income distribution is now available for several countries in the Organisation for Economic Development and Cooperation in the mid-1980s and for Great Britain from 1961 to 1991. The use of these data also casts doubt on the hypothesis that inequalities in the distribution of income are closely associated with variations in average life expectancy at birth among the richest nations of the world. PMID- 7496241 TI - Infected right atrial thrombus presenting as pulmonary embolism complicating central venous cannulation. PMID- 7496242 TI - ABC of medical computing. Keeping your computer healthy and legal. PMID- 7496243 TI - Nurses may make surgical house officers obsolete. PMID- 7496244 TI - Survey illustrates misconception of the Calman proposals. PMID- 7496245 TI - Junior doctors' hours. Flexible partial shifts can work. PMID- 7496246 TI - Harmonising specialist training in Europe. PMID- 7496247 TI - Junior doctors' hours. Flexible partial shifts should be made to work. PMID- 7496248 TI - Summative assessment for general practitioner registrars. System is currently used in west of Scotland. PMID- 7496249 TI - NRPB will provide advice on radiation. National Radiological Protection Board. PMID- 7496250 TI - Summative assessment for general practitioner registrars. Summative assessment will protect patients from incompetent GPs. PMID- 7496251 TI - Bone mineral densitometry in clinical practice. Differences in reference values are important. PMID- 7496253 TI - Comparing hospital and chiropractic treatment for back pain. Trial did not compare like with like. PMID- 7496252 TI - Bone mineral densitometry in clinical practice. Different guidelines will yield different estimates of prevalence. PMID- 7496254 TI - Comparing hospital and chiropractic treatment for back pain. Results were clinically trivial. PMID- 7496255 TI - Practical pulse oximetry. Overnight oximetry is easy and useful. PMID- 7496256 TI - Practical pulse oximetry. Saturation can be normal with dangerously high pCO2. PMID- 7496257 TI - Advance directives increase personal autonomy. PMID- 7496258 TI - Government health warnings may encourage adolescents to smoke. PMID- 7496259 TI - Misleading meta-analysis. One incorrect meta-analysis does not invalidate them all. PMID- 7496260 TI - Misleading meta-analysis. Subject to many potential biases. PMID- 7496261 TI - Exercise responses in the chronic fatigue syndrome. Objective assessment of study is difficult without knowledge of data. PMID- 7496262 TI - Volume and mortality in coronary artery bypass grafting. PMID- 7496263 TI - Medical library innovates on the Internet. PMID- 7496264 TI - Nurse triage increases emergency department waiting times. PMID- 7496265 TI - HTLV-I screening in Britain. PMID- 7496266 TI - New incentives for general practitioners in London. PMID- 7496267 TI - Outpatient follow up. PMID- 7496268 TI - Psychosocial interventions in cancer. PMID- 7496269 TI - Registering patients and paying capitation in family practice. PMID- 7496270 TI - Czech doctors threatened with prosecution. PMID- 7496271 TI - US poised to outlaw late abortion technique. PMID- 7496274 TI - Ban on stage hypnotism is not needed, says report. PMID- 7496273 TI - Tobacco sales soar in developing countries. PMID- 7496272 TI - British ministry condemned for Gulf war response. PMID- 7496276 TI - Dutch appeal court dismisses case against doctor. PMID- 7496275 TI - Britain restricts lice treatment. PMID- 7496277 TI - Israel condemned for torture by shaking. PMID- 7496278 TI - Patients with hip fractures need better care, says report. PMID- 7496279 TI - Overseas doctors face new rules to work in Britain. PMID- 7496280 TI - Psychosis with good prognosis in Afro-Caribbean people now living in the United Kingdom. AB - OBJECTIVES: To compare the course and outcome of psychotic illness in a group of Afro-Caribbean patients resident in the United Kingdom and a group of white British patients. DESIGN: Cohort study of consecutive admissions followed up for four years. SUBJECTS: 113 patients with psychotic illness of recent onset admitted to two south London hospitals. MAIN OUTCOME MEASURES: Course of illness, history of self harm, social disability, treatment received, and hospital use adjusted for socioeconomic origin. RESULTS: The Afro-Caribbean group spent more time in a recovered state during the follow up period (adjusted odds ratio 5.0; 95% confidence interval 1.7 to 14.5), were less likely to have had a continuous illness (0.3; 0.1 to 0.8), were less at risk of self harm (0.2; 0.1 to 0.8), and were less likely to have been prescribed antidepressant treatment (0.3; 0.1 to 0.9). There were no differences in hospital use, but the Afro-Caribbean group had more involuntary admissions (8.9; 2.1 to 35.6) and more imprisonments over the follow up period (9.2; 1.6 to 52.3). CONCLUSIONS: Afro-Caribbean patients in the United Kingdom have a better outcome after psychiatric illness than do white people. The combination of high incidence and more benign course of illness of psychotic illness in this group may be due, at least in part, to a greater exposure to precipitants in the social environment. PMID- 7496281 TI - Cognitive behavioural therapy for medically unexplained physical symptoms: a randomised controlled trial. AB - OBJECTIVE: To examine the additional effect of cognitive behavioural therapy for patients with medically unexplained physical symptoms in comparison with optimised medical care. DESIGN: Randomised controlled trial with follow up assessments six and 12 months after the baseline evaluation. SETTING: General medical outpatient clinic in a university hospital. SUBJECTS: An intervention group of 39 patients and a control group of 40 patients. INTERVENTIONS: The intervention group received between six and 16 sessions of cognitive behavioural therapy. Therapeutic techniques used included identification and modification of dysfunctional automatic thoughts and behavioural experiments aimed at breaking the vicious cycles of the symptoms and their consequences. The control group received optimised medical care. MAIN OUTCOME MEASURES: The degree of change, frequency and intensity of the presenting symptoms, psychological distress, functional impairment, hypochondriacal beliefs and attitudes, and (at 12 months of follow up) number of visits to the general practitioner. RESULTS: At six months of follow up the intervention group reported a higher recovery rate (odds ratio 0.40; 95% confidence interval 0.16 to 1.00), a lower mean intensity of the physical symptoms (difference -1.2; -2.0 to -0.3), and less impairment of sleep (odds ratio 0.38; 0.15 to 0.94) than the controls. After adjustment for coincidental baseline differences the intervention and control groups also differed with regard to frequency of the symptoms (0.32; 0.13 to 0.77), limitations in social (0.35; 0.14 to 0.85) and leisure (0.36; 0.14 to 0.93) activities, and illness behaviour (difference -2.5; -4.6 to -0.5). At 12 months of follow up the differences between the groups were largely maintained. CONCLUSION: Cognitive behavioural therapy seems to be a feasible and effective treatment in general medical patients with unexplained physical symptoms. PMID- 7496282 TI - Success of cardiopulmonary resuscitation after heart attack in hospital and outside hospital. AB - OBJECTIVES: To determine factors associated with cardiopulmonary resuscitation being attempted after cardiac arrest from myocardial infarction, in or outside hospital, and estimate short term and long term survival rates. DESIGN: Descriptive cross sectional and cohort study. SETTING: Community based register of all suspected heart attacks and sudden cardiac deaths in Lower Hunter region of New South Wales, Australia. SUBJECTS: 4924 men and women aged 25-69. MAIN OUTCOME MEASURES: Rates of attempted cardiopulmonary resuscitation and survival after successful resuscitation. RESULTS: Cardiopulmonary resuscitation was attempted in 41% of cases of cardiac arrest after myocardial infarction outside hospital and 63% of cases in hospital. Survival rates at 28 days were 12% and 39% respectively. Among the survivors, although 41% had another myocardial infarction (or coronary death), 81% of both groups were still alive two years later. Younger and better educated people were more likely to receive cardiopulmonary resuscitation in either setting, and being married predicted cardiopulmonary resuscitation being attempted outside hospital. Younger age predicted better survival rates after attempted resuscitation in hospital. CONCLUSIONS: The reasons for better education to predict cardiopulmonary resuscitation being attempted need explanation. The higher survival rate after cardiopulmonary resuscitation in hospital compared with outside hospital and the good long term prognosis for survivors in both settings suggest that attempts to improve success of cardiopulmonary resuscitation outside hospital may be worth while. PMID- 7496283 TI - Epidural anaesthesia and low back pain after delivery: a prospective cohort study. AB - OBJECTIVE: To determine whether epidural anaesthesia during labour and delivery is a risk factor for postpartum back pain. DESIGN: Prospective cohort study with follow up at one day, seven days, and six weeks after delivery. SETTING: Teaching hospital in Montreal. SUBJECTS: 329 women who delivered a live infant(s) during the study period. Exclusion criteria were back pain before pregnancy and delivery by elective caesarean section. INTERVENTION: Epidural anaesthesia during labour and delivery. MAIN OUTCOME MEASURES AND RESULTS: The primary outcome variable was development of postpartum low back pain. Back pain was quantified with self reports (yes/no), a pain score (numeric rating scale), and degree of interference with daily activities. Of the 329 women, 164 received epidural anaesthesia during labour and 165 did not. The incidence of low back pain in epidural v non-epidural group was 53% v 43% on day one; 21% on day seven; and 14% v 7% at six weeks. The relative risk for low back pain (epidural v non-epidural) adjusted for parity, delivery, ethnicity, and weight was 1.76 (95% confidence interval 1.06 to 2.92) on day one; 1.00 (0.54 to 1.86) on day seven; and 2.22 (0.89 to 5.53) at six weeks. There were no differences between the two groups in pain scores or the frequency of interference with daily activities. Similar results were obtained in the subgroup of women with low back pain of new onset--that is, those women with no back pain during their pregnancy. CONCLUSIONS: Postpartum low back pain was common but decreased considerably over the short term. The association between epidural anaesthesia and postpartum low back pain was inconsistent over time with a significantly increased risk of low back pain (epidural v non-epidural) noted only on the first day after delivery. PMID- 7496284 TI - Prospective observational study to assess value of prostate specific antigen as screening test for prostate cancer. AB - OBJECTIVE: To evaluate measurement of serum prostate specific antigen as a potential screening test for future clinical prostate cancer among healthy men. DESIGN: Nested case-control study with stored serum samples collected from 49,261 men with follow up using national death and cancer registration systems. SUBJECTS: 265 asymptomatic men who subsequently developed clinical prostate cancer and 1055 controls matched for age, study centre, and duration of storage of samples. MAIN OUTCOME MEASURES: Distribution of concentrations of the antigen in men who developed prostate cancer and in controls. RESULTS: Prostate specific antigen concentrations were significantly higher in men who subsequently developed prostate cancer than in controls. In the first three years after blood collection the median concentration was 23 times greater in cases than in controls of the same age at the same centre (that is, 23 multiples of the median). A smaller difference persisted thereafter; 4.0 multiples of the median 3 6 years after blood collection, 3.6 6-10 years, and 1.8 after 10 years. In the first three years the proportion of men who developed prostate cancer and had raised levels of the antigen (> or = 12 multiples of the median) (detection rate or sensitivity) was 81% (95% confidence interval 54% to 96%). The proportion of men who did not develop prostate cancer but had levels this high (false positive rate) was only 0.5%. CONCLUSION: Prostate specific antigen measurement is a highly discriminatory screening test for prostate cancer among healthy men. In the general population, 60-74 year old men who had > or = 12 times the normal median level would have about a 50% chance of developing clinical prostate cancer in the next three years. Measurement of this antigen is a good enough screening test to justify a randomised controlled trial to determine any reduction in mortality from prostate cancer. PMID- 7496286 TI - Routine surgical follow up: do surgeons agree? PMID- 7496285 TI - Increased mortality among Dutch development workers. PMID- 7496288 TI - Prospective seroepidemiological evidence that human papillomavirus type 16 infection is a risk factor for oesophageal squamous cell carcinoma. PMID- 7496287 TI - Increased risk of diarrhoea caused by Clostridium difficile in elderly patients receiving cefotaxime. PMID- 7496289 TI - Alterations in prescribing by general practitioner fundholders: an observational study. AB - OBJECTIVES: To compare prescribing in general practices before and after they become fundholders to assess whether this affected prescribing patterns. DESIGN: Analysis of prescribing data (PACT) for one year before and one year after practices become first, second, or third wave fundholders and comparison with practices that were not fundholders during any part of the study. MAIN OUTCOME MEASURES: Prescribing costs (net ingredient cost per prescribing unit), prescribing volume (items per 1000 prescribing units), net ingredient cost per item, and percentage of generic prescribing. SETTING: Former Mersey Regional Health Authority. SUBJECTS: 100 fundholders (20 first wave, 31 second wave, 49 third wave) and 312 nonfundholders. RESULTS: Prescribing costs and volume rose throughout the study in all groups. In all three fundholding waves the rate of increase of prescribing costs was significantly lower than for nonfundholders. Both cost per item and prescribing volume tended to decrease, the former probably because of a significant increase in generic prescribing. Fundholding and non fundholding practices differed in several respects. CONCLUSION: Fundholding has altered practice prescribing patterns compared with those of nonfundholders, increasing generic prescribing and reducing the rate of increase of prescribing costs. PMID- 7496291 TI - An evidence based approach to individualising treatment. AB - To which groups of patients can the results of clinical trials be applied? This question is often inappropriately answered by reference to the trial entry criteria. Instead, the benefit and harm (adverse events, discomfort of treatment, etc) of treatment could be assessed separately for individual patients. Patients at greatest risk of a disease will have the greatest net benefit as benefit to patients usually increases with risk while harm remains comparatively fixed. To assess net benefit, the relative risks should come from (a meta-analysis of) randomised trials; the risk in individual patients should come from multivariate risk equations derived from cohort studies. However, before making firm conclusions, the assumptions of fixed adverse effects and constant reduction in relative risk need to be checked. PMID- 7496290 TI - Hepatology. PMID- 7496292 TI - Occult intracranial tumours masquerading as early onset anorexia nervosa. PMID- 7496294 TI - Avoiding drug errors. Adopting new NHS specification helps. PMID- 7496295 TI - Avoiding drug errors. Read the label. PMID- 7496293 TI - ABC of atrial fibrillation. History, epidemiology, and importance of atrial fibrillation. PMID- 7496296 TI - Avoiding drug errors. Reporting of errors should be free of recrimination. PMID- 7496297 TI - Avoiding drug errors. Pharmacists and computerised prescribing can help. PMID- 7496298 TI - Avoiding drug errors. Inappropriate prescribing is "treatable". PMID- 7496299 TI - Avoiding drug errors. Reporting of critical incidents shows how errors occur. PMID- 7496300 TI - Preventing local complications of depot neuroleptics. PMID- 7496301 TI - Suicide after discharge from psychiatric hospitals in Scotland. PMID- 7496302 TI - Clinical efficacy of treatment for head lice. Counting head lice by visual inspection flaws trials' results. PMID- 7496303 TI - Clinical efficacy of treatment for head lice. Authors differ on assessment of flaws in trials. PMID- 7496304 TI - Elective ventilation is cost effective and humane. PMID- 7496306 TI - Low back pain. PMID- 7496307 TI - Decision analysis and screening for Down's syndrome. Estimate of uptake of amniocentesis is overoptimistic. PMID- 7496305 TI - Public lacks knowledge about genetic testing and gene therapy. PMID- 7496309 TI - Decision analysis and screening for Down's syndrome. Costs were overestimated. PMID- 7496310 TI - Decision analysis and screening for Down's syndrome. Testing should be in all women. PMID- 7496311 TI - Right to choose. PMID- 7496308 TI - Decision analysis and screening for Down's syndrome. Not using age specific values invalidates study. PMID- 7496312 TI - [The effect of 5-fluorouracil on the clonogenic hematopoietic cells (CFU-S) of embryonic and sexually mature mice]. AB - We studied sensitivity of various types of clonogenic hemopoietic cells (CFU-S-7, CFU-S-11, and CFU-ep) on the liver of 14-days embryos and from the bone marrow and spleen of adult mice to the cytotoxic agent 5-fluorouracil (5-FU) in vitro and in vivo. We discovered that different types of CFU-S of the embryos and adult mice has similar sensitivity to 5-FU in vitro. At the same time in vivo the bone marrow CFU-S-7 display a higher sensitivity to 5-FU than CFU-S-11, thus agreeing with the published data (Hodgson and Bradley, 1979). The differences between the 5-FU effects in vitro and in vivo are related to the hemopoietic microenvironment, which modulates the cytotoxic 5-FU effect in vivo. PMID- 7496313 TI - [Trace elements in the neurophysiological mechanisms of the development of interhemispheric asymmetry]. AB - The content of microelements in the cerebral cortex of rats featuring motor asymmetry was studied by the method of emission spectral analysis with inductively bound argon plasma. The content of microelements (Cd, Co, Cu, Fe, Mn, Mo, P, S, and Zn) differs reliably in the subdominant and dominant hemispheres. We propose that interhemispheric asymmetry of the content of microelements plays the key role in formation of the brain functional asymmetry. PMID- 7496314 TI - [The cytogenetic effect of ultralow doses of nitrosomethylurea]. AB - Cytogenetic effects of ultra low doses of nitrosomethylurea in the range 10(-12) 10(-17) mol/(kg x day) on the chromosome structure were studied in the Ehrlich tumor and leucosis L-12110 cells. It was shown that nitrosomethylurea at an ultra low dose of 10(-17) mol/(kg x day) induces chromosome aberrations in the cells of the both studied tumors after a single and multiple injections to random bred and linear animals. The recorded effect suggests that with the decrease of therapeutic dose by 13 orders of magnitude the cytogenetic activity of the drug decreases by no more than one order of magnitude. PMID- 7496316 TI - [The taxonomic diversity of the rodents within the limits of the Asian ecological transect]. AB - The diversity of rodents is considered at various taxonomic levels within the limits of the Asian ecological transect, which crosses Asia from the Taimyr Peninsula in the north to the southern coast of Indostan and embraces territories of seven countries: Russia, Kazakhstan, Kyrgyzstan, Mongolia, China, Nepal, and India. The highest diversity of rodents is observed in the south of West Siberia and in mountains of Middle Asia. The least number of all taxa was noted in the Taimyr Peninsula and in the Tibet region. PMID- 7496317 TI - [A pharmacokinetic study of para-aminobenzoic acid administered into the rabbit eye by phonophoresis]. AB - The pharmacokinetics of para-aminobenzoic acid (PABA) was studied after introduction of 3H-PABA in the rabbit eye by phonophoresis (1.8 ml 3H-PABA per eye, calculated from 10 muCi in 1 ml 0.9% sodium chloride). The label was found by liquid scintillation in all studied eye tissues, including the retina. Within 1 h the label content in the tissues was higher than within 3 after the isotope introduction. Thus, in addition to subcutaneous injection (Stroeva et al., 1992), another method of PABA introduction in the eye was used in order to further study its activity as a medicinal drug during treatment of degenerative processes in the retina. PMID- 7496319 TI - [The effect of the incomplete terminal repair of the linear double-stranded DNA molecule]. AB - A possible effect of incomplete terminal DNA repair providing for shortening of telomeric DNA in the somatic cells, specifically postmitotic neurons, has been discussed. The effect of incomplete terminal repair can and must show up independently from the effect of incomplete terminal DNA replication proposed earlier. PMID- 7496318 TI - [The 5-fluorouracil sensitivity of the hematopoietic stroma of the bone marrow and spleen]. AB - We studied the effect of a single injection of 5-fluorouracil (5-FU) (150 mg/kg) on the numbers of stromal precursor cells (CFU-F) in the bone marrow and spleen and their capacity for formation of hemopoietic foci after ectopic transplantation under the renal capsule. 5-FU decreased the number of CFU-F in the bone marrow and spleen of mice and rats to 30 - 50% of the control values. The size of transplants, as estimated according to the number of hemopoietic clonogenic (CFU-S) and nucleated cells, diminished within the first two months after transplantation. Within four months after transplantation the size of the bone marrow transplants approached the control level, while that of the spleen transplants markedly exceeded the control level. Differences in the growth rate of regenerating transplants of the hemopoietic organs are discussed with special reference to the organ specific features of populations of the stromal cells. PMID- 7496315 TI - [An experimental in-vitro study of the effect of paraorbital mesenchyme with the Mi wh/Mi wh mutant genotype on the melanogenesis of the retinal pigment epithelium in mouse embryos]. AB - The effects of paraorbital +/+ and Mi wh/Mi wh mesenchyme (PM) on melanogenesis of the retinal pigment epithelium (RPE) of mouse embryos. The eye anlagen of 10 day-old +/+ and Mi wh/Mi wh mouse embryos were co-cultured in vitro during eight days. All eye anlagen of +/+ genotype without PM cocultured with anlagen of +/+ genotype with PM developed a pigmentation of RPE. However, the eye anlagen of +/+ genotype without PM and the eye anlagen of Mi wh/Mi wh genotype with PM when co cultured did not develop pigmentation of RPE. These results indicate that mutant allele Mi wh of locus mi changes the properties of PM which in mutant embryos does not exert the normal melanogenesis-promoting effect in relation to RPE. PMID- 7496320 TI - [The role of incomplete terminal repair of chromosomal DNA in the aging of neurons and postmitotic organisms]. AB - Incomplete repair of the DNA double helix terminus is an independent mechanism of the telomeric DNA shortening in non-dividing cells. This effect of incomplete terminal DNA repair may initiate aging of postmitotic neuroendocrine cells or even whole postmitotic organisms, if loss of the buffer telomeric DNA (due to incomplete terminal repair) is followed by the loss of significant information in the nucleotide sequences. We also consider a possibility of local aging of a cell group, e.g., in the locomotor system where extreme physical load favors accumulation of nuclear factors inducing accelerated shortening of telomeric DNA. PMID- 7496321 TI - Introduction: dietary fibre, butyrate and colorectal cancer. PMID- 7496322 TI - Epidemiology of fibre, resistant starch and colorectal cancer. PMID- 7496323 TI - Bacterial fermentation of complex carbohydrate in the human colon. PMID- 7496324 TI - The short-chain fatty acid butyrate induces apoptosis in colorectal tumour cell lines. PMID- 7496325 TI - Butyrate and markers of neoplastic change in the colon. PMID- 7496326 TI - Butyrate and colonic cytokinetics: differences between in vitro and in vivo studies. PMID- 7496328 TI - Cancer mortality in Europe, 1990-92. AB - Histograms of all age-standardized (world population) death certification rates for 23 cancers or groups of cancers for the period 1990-92 were produced for 35 countries of the European region (including a dozen new national entities) providing data to the World ealth Organization database. Substantial variations were observed in mortality from most common sites. For lung cancer the rate in males was 81/100,000 in Hungary, followed by Belgium, the Czech Republic, the Russian Federation and Poland, while in Sweden, Iceland and Norway, where comprehensive antismoking campaigns have been adopted over the last two decades, the rates were between 24 and 30 per 100,000 males. The lung cancer epidemic in European females is still in its early phases in most countries, with the sole exception of Scotland (29/100,000, ie the highest rates in the world), the rest of the UK, Denmark, Iceland, Ireland and Hungary. With reference to colorectal cancer, the highest rates were in the Czech Republic (38/100,000 males, 21/100,000 females) and other central European countries, and the lowest in Greece, Romania and a few Republics of the former Soviet Union, as well as Finland and Sweden. The highest gastric cancer mortality rates were in the Russian Federation (41/100,000 males, 18/100,000 females), followed by a few Republics of the former Soviet Union and Portugal in Western Europe. The highest breast cancer rates (25-29 per 100,000 females) were in the UK, Belgium, Ireland, The Netherlands, Denmark and other Scandinavian countries. For overall cancer mortality, the range of variation was between 260/100,000 in Hungary and 132/100,000 in Sweden for males, and between 142/100,000 in Denmark and 76/100,000 in Kyrgizstan for females, ie approximately a twofold variation in both sexes. PMID- 7496327 TI - The origins of Hodgkin's disease. AB - Despite significant advances in treatment, the aetiology of Hodgkin's disease has remained elusive. While epidemiology has uncovered the likely infectious nature of the malignancy, neither the causative agent nor the cell of origin have been identified. The available epidemiological, histological and molecular biological evidence were reviewed and the following conclusions reached. Hodgkin's disease is probably an infectious, but not contagious disease, and is compounded by immunodeficiency. The suspected role of the Epstein-Barr virus (EBV) as causative agent has been substantially weakened, although a role as co-factor is likely. A prime candidate for the cell of origin seems the interdigitating reticulum cell (IRC). Reed-Sternberg cells found in patients with Hodgkin's disease may represent in vivo hybridomas of the IRC with B- and/or T-cells. As IRC is unlikely to be susceptible to EBV, a retrovirus may be the culprit. Hybridomas may be formed when retroviral antigens expressed by a macrophage cell attract reactive B- and T-cells and, instead of an immune reaction, fusion occurs. Further research is necessary in the search for the causative agent of Hodgkin's disease. PMID- 7496330 TI - Egg consumption and risk-markers for colorectal neoplasia. AB - A strong positive relationship between dietary intake of eggs and risk of colorectal cancer has been observed in a number of epidemiological surveys. In this study we investigated the relationship between egg consumption and intermediate biomarkers of crypt cell proliferation in the colon and rectum of 75 patients who had adenomatous polyps or no evidence of bowel disease. Biopsies of normal flat mucosa were obtained at colonoscopy, and microdissected crypts were used to measure crypt length, frequency of mitosis per crypt and spatial distribution of mitoses within the crypt. There was no significant difference in the consumption of eggs by patients with adenomatous polyps (n = 53) and those without (n = 22). There was no significant positive correlation between frequency of egg consumption and any of the parameters of crypt cell proliferation in the group as a whole, nor when the patients with polyps were analysed separately. This study provides no biological evidence of any relationship between egg consumption and abnormal cell proliferation among patients at relatively high risk of colorectal neoplasia. PMID- 7496329 TI - Physical activity and survival from breast cancer. AB - The association between recreational physical activity and survival from breast cancer was investigated in a population-based cohort of breast cancer patients in Adelaide, South Australia. These patients, aged 20-74 years at diagnosis, had been recruited between 1982 and 1984 into a case-control study of diet, hormones and breast cancer. Of the 451 patients with breast cancer originally enrolled, 412 were followed for a median interval of 5.5 years. The study participants reported their weekly levels of light, moderate and vigorous recreational physical activity, which were then converted into estimates of kilocalories per week of energy expenditure. Despite some fluctuation in the hazard ratios by level of physical activity, there was little evidence for an association between total recreational physical activity and risk of death from breast cancer overall, or within menopausal strata. Similarly, there was little evidence for associations between individual types of recreational physical activity and risk of death from breast cancer. PMID- 7496331 TI - Superoxide dismutase and lipid-bound sialic acid in sera from children with cancers and juvenile chronic arthritis. AB - Both experimental and epidemiological studies suggest that patients with autoimmune and other chronic inflammatory diseases are more prone to develop cancer. The aim of the present investigation is to find some typical differences between the content of lipid-bound sialic acid (LSA) and the activity of superoxide dismutase (SOD) in sera from children with neoplastic diseases and juvenile chronic arthritis (JCA). Our results show 30% lower SOD activity in the sera from children with cancer compared with the sera from children with JCA. LSA levels 102% and 166% higher than in children with JCA were observed in the sera from children with blood and solid cancers, respectively. The relation LSA/SOD is about fivefold higher in children with cancers. A negative correlation (r = 0.720, P < 0.001) exists between LSA and SOD in sera from children with cancers. No such correlation was established in the group of children with JCA. We suppose that such differentiation disappears in the beginning of neoplastic process during prolonged therapy of autoimmune diseases. From our findings SOD and LSA appear to be putative markers of malignant disease with potential usefulness not only in JCA but also in other conditions associated with an increased risk of neoplastic development. PMID- 7496332 TI - Mass screening for colorectal cancer: statement of the European Group for Colorectal Cancer Screening. PMID- 7496333 TI - Exposure to extremely low frequency electromagnetic fields and the risk of malignant diseases--an evaluation of epidemiological and experimental findings. AB - Epidemiological and experimental studies concerning extremely low frequency electromagnetic field exposure and malignant diseases published up to 1 July 1994 were evaluated to assess the possible carcinogenicity of electromagnetic fields and the scientific basis for environmental and occupational standard setting. We concluded that there are possible associations between (i) an increased risk of leukaemia in children and the existence of, or distance to, power lines in the vicinity of their residence, (ii) an increased risk of chronic lymphatic leukaemia and occupational exposure to low frequency electromagnetic fields and (iii) an increased risk of breast cancer, malignant melanoma of the skin, nervous system tumours, non-Hodgkin lymphoma, acute lymphatic leukaemia or acute myeloid leukaemia and certain occupations. There is no scientific basis for occupational or environmental standard setting for low frequency electric or magnetic fields. PMID- 7496334 TI - Indocyanine green angiography in traumatic choroidal rupture: clinicoangiographic case reports. AB - Blunt trauma to the eye can cause choroidal ruptures. Often localization is obscured by hemorrhages, edema, and pigmentary changes. The use of indocyanine green (ICG) angiography to supplement fluorescein angiography in visualizing retinal and choroidal tissues was investigated. Three patients with acute traumatic choroidal ruptures received a retinal examination, including fluorescein and ICG angiography study. Choroidal ruptures were well localized in ICG angiograms in both early and late phases. In fluorescein angiograms, the defects were underestimated in early phases and overestimated in the later phases. In one patient with severe traumatic changes, ophthalmoscopy and fluorescein angiography identified all superficial splits, whereas ICG angiography readily detected deeper changes as well. ICG angiography is helpful in diagnosis major and minor ruptures of the choriocapillaris and the choroid, in defining the extent of traumatic ruptures, and in furthering the understanding of the pathology of traumatic ruptures, and in furthering the understanding of the pathology of traumatic tears. PMID- 7496335 TI - Insulin-like growth factor I (IGF-I) receptor/binding protein in human diabetic epiretinal membranes. AB - Insulin-like growth factor I (IGF-I) is thought to play a role in the development of proliferative diabetic retinopathy. Proliferative diabetic retinopathy is characterised by the formation of fibrovascular epiretinal membranes, and IGF-I may initiate and/or potentiate this epiretinal proliferation. To evaluate further the part played by IGF-I in the development of epiretinal tissue, we investigated the presence of IGF-I receptor/binding protein in proliferating diabetic fibrovascular epiretinal membranes. Five fibrovascular epiretinal membranes were obtained by vitrectomy from five patients with proliferating diabetic retinopathy. The presence of IGF-I receptors was investigated by autoradiography using 125I-labeled IGF-I on frozen sections. To characterise binding specificity, some sections were preincubated with either insulin or unlabeled IGF-I. Sections of post-mortem liver were used as controls. Strong labeling of cells with 125I labeled IGF-I was observed in all epiretinal membranes and in liver cells. Almost no autoradiographic labeling was observed in sections that had been blocked with non-radioactive IGF-I, and very little labeling was found following blockage with insulin. Our preliminary study suggests the presence of IGF-I receptor/binding protein in human diabetic epiretinal membranes. These results support the hypothesis that IGF-I may be involved in the formation of proliferative diabetic membranes. PMID- 7496337 TI - Posterior drainage of intraocular fluid: an experimental approach on bovine cadaver eyes. AB - We evaluated the posterior drainage of saline in cadaver bovine eyes using different experimental procedures. The saline perfusion via a pars plana infusion into the vitreous cavity was measured in the following 5 different groups of 20 enucleated bovine eyes: 1, control; 2, vitrectomy; 3, vitrectomy + retinectomy; 4, vitrectomy + retinectomy + removal of retinal pigment epithelium (RPE); and 5, vitrectomy + retinectomy + removal of RPE + choroidectomy. The mean values obtained for the facility of saline outflow were as follows group 1, 0.0458 microliter min-1 mmHg-1; group 2, 0.0393 microliter min-1; group 3, 0.1308 microliter min-1 mmHg-1; group 4, 0.2288 microliter min-1; and group 5, 0.9985 microliter/min-1 mmHg-1. The retina appeared to be the major barrier to fluid movement from the vitreous to the chorioid, perhaps due to the lack of extracellular space and to the tight and impervious structure of the retina. The barrier function of the retina may explain the hypotony seen after rhegmatogenous retinal detachment and the decrease in intraocular pressure observed after the removal of silicone oil in eyes that had undergone retinectomy. Our results suggest that the reduction in intraocular pressure occurring after retinectomy is attributable to the new drainage pathway. PMID- 7496336 TI - Intercellular adhesion molecule-1 levels in plasma and vitreous from patients with vitreoretinal disorders. AB - Intercellular adhesion molecule 1 (ICAM-1) is a member of the immunoglobulin superfamily with important functions in immune activation and inflammation. Its interaction with different cytokines [interleukin 1 (IL-1), tumor necrosis factor alpha (TNF-alpha), interferon-gamma (IFN-gamma)] is important for lymphocyte migration into inflammatory sites. We used a sandwich enzyme immunoassay (EIA) for the quantitative determination of soluble ICAM-1 (cICAM-1) in vitreous and plasma from patients undergoing vitrectomy for a variety of proliferative vitreoretinal disorders. The values obtained were compared with the total vitreal protein. The respective concentrations of cICAM-1 in vitreous were as follows control samples, 3.47 +/- 1.84 ng/ml; proliferative diabetic retinopathy (PDR) of diabetes type I 27.43 +/- 14.72 ng/ml; PDR of diabetes type II, 32.46 +/- 10.31 ng/ml; idiopathic proliferative vitreoretinopathy 35.74 +/- 15.30 ng/ml; and traumatic PVR, 45.23 +/- 24.24 ng/ml. Plasma samples yielded the following concentrations: controls, 415 +/- 43.4 ng/ml; PDR of diabetes type I, 469 +/- 96.9 ng/ml; PDR of diabetes type II, 425 +/- 65.4 ng/ml; idiopathic PVR, 402 +/- 119.9 ng/ml; and traumatic PVR, 434 +/- 118.6 ng/ml. Our results demonstrate high levels of ICAM-1 in most proliferative vitreoretinal disorders. In PDR and in traumatic PVR, cICAM-1 levels were elevated significantly more than were total vitreal protein levels. In traumatic PVR, patients with a short interval between previous surgery or traumatic event demonstrated the highest levels of cICAM. Since plasma levels were not significantly altered, we suggest that local cICAM-1 production, possibly from macrophages, may be of importance in the early phase of PVR and PDR by enhancing immune activation and inflammation. PMID- 7496338 TI - Retinal hemodynamics in patients with chronic open-angle glaucoma. AB - Recently it has been demonstrated that retinal hemodynamics are disturbed in patients with chronic open-angle glaucoma. As the underlying cause a reduction in perfusion pressure due to increased intraocular pressure (IOP) and deficiencies of retinal autoregulation has been discussed. The present study was undertaken to clarify the influence of filtering surgery on retinal hemodynamics in patients with chronic open-angle glaucoma. A total of 17 patients with chronic open-angle glaucoma aged between 37 and 86 years were included in this prospective study. All patients underwent digital fluorescein angiography before and 10 days after fistulating procedures. From the angiograms the arteriovenous passage time (AVP) and arterial mean dye-bolus velocity (MDV) were quantified by means of digital picture analysis. At baseline the AVP was significantly prolonged in the patients as compared with reference values (AVP, 2.5 +/- 0.8 versus 1.6 +/- 0.4 s; P < 0.01). After the fistulating procedure (IOP: before, 29 +/- 5 mmHg; after, 16 +/- 4 mmHg) the AVP was significantly reduced as compared with baseline values (AVP, 2.5 +/- 0.8 versus 2.0 +/- 0.4 s; P < 0.05), whereas the MDV showed only a slight increase (MDV, 5.70 +/- 0.89 versus 5.99 +/- 0.92 mm/s; P > 0.05). This study confirms a disturbance of retinal hemodynamics in patients with chronic open angle glaucoma. The significant reduction in AVP observed after lowering of the IOP by fistulating procedures demonstrates the positive influence of IOP reduction on the retinal circulation. The improvement in retinal circulation may prevent the occurrence of further glaucomatous damage after fistulating procedures. PMID- 7496340 TI - The Tubingen Glaucoma Study. Glaucoma filtering surgery--a retrospective long term follow-up of 254 eyes with glaucoma. AB - A major focus of our study was the ability to predict the long-term success of filtering surgery. The results of glaucoma filtering surgery (trabeculectomy) as performed by the staff of the University Eye Hospital of Tubingen from 1988 through 1994 on 254 eyes of 214 patients are presented, with particular emphasis being placed not only on intraocular pressure (IOP) control but also the progression of glaucomatous damage (visual field loss or disc damage) and the etiology of visual acuity losses. There was an overall success rate of 64%. Rigid criteria for success included an IOP of less than 30 mmHg, no further visual field loss or disc damage, and no glaucomatous etiology for a decrease in visual acuity. A total of 35 eyes (13.8%) showed a loss of visual acuity after 6 months amounting to 2 or more Snellen lines, caused mainly by lens opacification, hypotony maculopathy, and "wipeout" (loss of the central visual field in the absence of another explanation). We found that there is a considerable risk for sudden loss of visual acuity after operation on older patients with small residual visual fields and severe hypotony on the 1st postoperative day. There was a 13% incidence of failure of blebs. A total of 34 eyes had an early IOP rise of more than 30 mmHg after surgery. A postoperative hypotony of less than 3 mmHg was more significantly seen in patients treated with 5-fluorouracil. Moreover, the results show that in some instances, a long-term use of topically applied glaucoma medication can adversely affect the results of fistulizing surgery.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7496339 TI - Development of a new Er: YAG laser conception for laser sclerostomy ab externo: experimental and first clinical results. AB - The erbium:YAG laser is a preferable energy source for laser sclerostomy ab externo due to the high absorption of its radiation (2940-nm wavelength) in the aqueous parts of scleral tissue. It has thus far been a disadvantage that laser energy has to be transmitted from the laser source to the application site via very susceptible special optical fibers (ZrF). As a consequence of technical improvements, the laser source could be integrated into the application probe. Therefore, only a robust quartz fiber was necessary for contact application. A fiber diameter of 400 microns was chosen according to the experience gained in previous studies. A total of 24 eyes with chronical open-angle glaucoma were treated. A fistula end-point detection unit was developed for safe indication of a full-thickness perforation of the sclera. With a fiber diameter of 400 microns, a postoperative success rate of about 54% was achieved at 6 months follow-up. The automated full-thickness perforation detection prevented the application of additional, surplus laser pulses in the anterior chamber. The postoperative success rate could be improved over that achieved in previous studies. The new device/conception offers more technical reliability and is a further step toward minimally invasive fistulating laser surgery of open-angle glaucoma. PMID- 7496341 TI - Delay in corneal allograft rejection due to anti-CD4 antibody given alone and in combination with cyclosporin A and leflunomide. AB - Anti-CD4 monoclonal antibodies (mAbs) have been shown to inhibit in vitro T-cell activation and proliferation to both antigens and mitogens. Animals studies have demonstrated the immunosuppressive potency of anti-CD4 mAbs given in vivo for therapy of autoimmune disease and following allografting. Similarly, leflunomide (LF), a new potent immunosuppressive, has been shown to be effective in preventing autoimmune disorders and reactions leading to organ transplant rejection. LF is thought to antagonize cytokine activity, thereby interfering with T-helper-cell-dependent B- and T-lymphocyte proliferation. A new anti-CD4 antibody (RIB 5/2) was investigated alone and in combination with cyclosporin A (CsA) and LF for the treatment of corneal allograft rejection in the rat. Corneal buttons were grafted from Lewis/Brown Norway rats to Lewis recipients. Animals were randomly assigned to the following treatment groups: (1) untreated; (2) CsA at 1.5 mg/kg; (3) RIB 5/2 at 2.5 mg/kg; (4) RIB 5/2 at 2.5 mg/kg and CsA at 1.5 mg/kg; (5) RIB 5/2 at 2.5 mg/kg, CsA at 1.5 mg/kg and LF at 10 mg/kg; (6) RIB 5/2 at 4 mg/kg; (7) RIB 5/2 at 4 mg/kg and CsA at 1.5 mg/kg; and (8) RIB 5/2 at 4 mg/kg, CsA at 1.5 mg/kg, and LF at 10 mg/kg. RIB 5/2 was given intraperitoneally at 24 h before surgery, on the day of grafting, and on postoperative day 1 and was continued every 2nd day until the rat had received ten doses in all (postoperative day 15).(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7496342 TI - Localized transscleral staphylococcal abscess simulating a neoplasm. AB - A 47-year-old woman with a history of breast cancer developed a focal epibulbar mass in the right eye. The initial diagnosis was nodular episcleritis, but the lesion failed to respond to local and systemic corticosteroids. The rule out an epibulbar metastatic carcinoma an excisional biopsy was performed, which demonstrate a localized staphylococcal abscess that had locally eroded the sclera and exposed the underlying uveal tract. Covering the defect with a scleral patch graft and promptly instituting oral amoxicillin/clavulanic acid and topical cefazolin treatment resulted in an uncomplicated recovery. Infectious scleral abscess in an uncommon entity and the differential diagnosis includes nodular episcleritis and epibulbar metastatic tumor. PMID- 7496343 TI - Ophthalmological tests in patients with keratoconjunctivitis sicca with and without association of primary Sjogren's syndrome. AB - We compared clinical, laboratory and histological tests in patients with keratoconjuctivities sicca with and without association of primary Sjogren's syndrome. Rosa bengal staining, osmolarity and impression cytology had high specificity and sensitivity for a correct diagnosis of keratoconjunctivitis sicca with and without primary Sjogren's syndrome. Schirmer and breakup time (BUT) tests were not quite as specific or sensitive. Impression cytology was the only ocular test that showed significant differences between the two pathological groups. PMID- 7496344 TI - Microperimetry in patients with central serous retinopathy. AB - In patients with acute central serous retinopathy (CSR), evaluation of visual acuity alone may not represent visual function. In patients with acute CSR, visual function may be disturbed by localized scotomas, distortion, and waviness. For the assessment of localized light sensitivity and stability of fixation, patients with CSR were evaluated by fundus perimetry with a scanning laser ophthalmoscope (SLO 101, Rodenstock Instruments). In all, 21 patients with acute CSR and 19 healthy volunteers were included in the study. Diagnosis of CSR was established by ophthalmoscopy and digital video fluorescein angiography. All patients and volunteers underwent static suprathreshold perimetry with the SLO. Light sensitivity was quantified by presenting stimuli with different light intensities (intensity, 0-27.9 dB above background; size, Goldmann III; wavelength, 633 nm) using an automatic staircase strategy. Stimuli were presented with simultaneous real-time monitoring of the retina. Fixation stability was quantified by measuring the area encompassing 75% of all points of fixation. Light sensitivity was 18-20 dB in affected areas, whereas in healthy eyes and outside the affected area, values of 22-24 dB were obtained. Fixation stability was significantly decreased in the affected eye as compared with normal eyes (33 +/- 12 versus 21 +/- 4 min of arc; P < 0.01). Static perimetry with an SLO is a useful technique for the assessment of localized light sensitivity and fixation stability in patients with macular disease. This technique could provide helpful information in the management of CSR. PMID- 7496346 TI - Macroaneurysm of a cilioretinal artery. PMID- 7496345 TI - Plasma concentrations of lidocaine after proximal and peripheral facial nerve block. AB - We compared plasma levels of lidocaine after the administration of proximal and peripheral blocks of the facial nerve in 30 cataract patients to evaluate the risk of systemic toxicity after both procedures. We performed the two block techniques with 5 ml 1% lidocaine solution plus (1:20,000) naphazoline nitrate as follows: the proximal block, at the dorsal rim of the mandible and the peripheral block, by subcutaneous infiltration of the lids. After the administration of the proximal block, mean plasma concentrations of lidocaine increased continuously for 15 min, whereas after the peripheral block they peaked within 12 min. Mean plasma concentrations were significantly lower following the proximal facial block than after the peripheral block of the facial nerve's terminal branches (P = 0.008-0.039, Wilcoxon test). In all patients, plasma concentration of lidocaine were below toxic levels. PMID- 7496348 TI - Proceedings of the 3rd GIMEMA Conference on Recent Advances in the Cytobiology of Acute Leukemias. Bari, Italy, 13-14 May 1994. PMID- 7496347 TI - Recommended procedures for the classification of acute leukaemias. AB - The classification of acute leukaemias is now widely based on a combined morphological, cytochemical and immunophenotyping approach. Difficulties are frequently encountered however in reaching an acceptable degree of diagnostic concordance between different laboratories because of variations in the techniques used (in terms of methodologies, reagents and equipment) and diagnostic interpretation. The International Council for Standardization in Haematology (ICSH) convened an expert panel to consider currently available diagnostic techniques with the aim of defining a minimum cytochemical and immunological diagnostic panel that could be used as core components for the classification of acute leukaemia. The proposed ICSH scheme, which attempts to balance the basic requirement for providing precise and informative diagnostic information without limiting its use to only those laboratories with sophisticated facilities, is based on three sequential levels of investigation; primary cytochemistry, intracellular phenotyping and membrane immunophenotyping. The minimum ICSH recommended cytochemistries comprise myeloperoxidase (MPO), chloroacetate esterase (ChlorE) and alpha-naphthyl acetate esterase (ANAE), and standardised methods for these cytochemistries are detailed in this communication. For cases of acute leukaemia that remain unclassified by primary cytochemistry, subsequent immunological analyses for cytoplasmic CD3, CD22, MPO and nuclear TdT are recommended. The ICSH panel considers that the use of these minimum primary cytochemical and intracellular phenotyping procedures will lead to the consistent classification of most acute leukaemias, and that the third level of investigation (membrane immunophenotyping) should be used for the purposes of confirmation, diagnostic clarification of atypical leukaemias, and the subtyping of acute lymphoblastic leukaemias (ALL). The ICSH panel also recognised that there are a number of additional technologies which can provide definitive diagnostic information, such as cytogenetics and DNA genotyping, but these were excluded from the minimum panel because of their restricted availability. While many specialised laboratories, particularly in the areas of diagnostic research, will continue to use individual investigatory protocols, it is considered that the inclusion of the ICSH scheme as core components would lead to greater consistency when comparing independent studies of acute leukaemia. PMID- 7496349 TI - Immunological definition of acute minimally differentiated myeloid leukemia (MO) and acute undifferentiated leukemia (AUL). AB - Immunophenotyping has become an important tool in the diagnosis of acute leukemia for several reasons. Indeed the use of a standardized panel of monoclonal antibodies (MoAb) to B and T cells, and myeloid cells, as well as non lineage restricted antigens, permits allocation of more than 98% of acute leukemia to their respective lineage. In ALL, immunophenotyping has established a basis for precise and biologically oriented classification of the disease which may be of prognostic importance. In AML immunological markers are particularly important for identification of acute leukemia with minimal myeloid, erythroblastic or megakaryoblastic differentiation. Immunological markers also allow the identification of acute leukemias with minimal or aberrant marker expression, acute biphenotypic leukemia in which single cells coexpress different lineage associated markers and acute bilineage leukemia where there are two separate blast cell populations (usually lymphoid and myeloid). There is sometimes confusion in the literature about the definition of acute unclassifiable and acute undifferentiated leukemia. This is mainly due to misinterpretation of phenotypic data or to the lack of relevant lineage specific markers in these studies, especially for the detection of cytoplasmic antigens. Indeed, it is important to stress that in hematopoietic precursors, antigens detected by monoclonal antibodies first appear in the cytoplasm during early differentiation and are only expressed on the membrane later. This has been demonstrated not only for the T lineage (Cy CD3), the B lineage (CyCD22) but also for the myeloid lineage (CyCD13).(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7496350 TI - Cytogenetics of hybrid acute leukemias. AB - Although the recognition of hybrid acute leukemia (HAL) is still controversial, several reports have described cytogenetic findings in these leukemias over the last 3 years. A distinct chromosomal profile appears to be associated with different immunologic subsets of HAL. The classical t(15;17), and inv(16) as well as abnormalities of the long arm of chromosome 5 and/or 7 are preferentially associated with acute myeloid leukemia (AML) with T-cell features; the t(8;21)(q22;q22), the Ph chromosome, and 11q23 rearrangements are more frequently found in AML with B-cell features; the Ph chromosome, t11q23 and 14q32 breaks without rearrangements of the immunoglobulin heavy chain gene may be associated with acute lymphoblastic leukemia (ALL) with myeloid markers. In addition, some chromosome aberrations may be encountered more frequently in acute leukemia with major phenotype deviations than in unselected cases of acute leukemia: namely the Ph chromosome, 11q23 rearrangements, and +13. These chromosome changes appear to be associated with a low complete remission (CR) rate. An association has been documented in some patients with ALL between the presence of the t(9;22) and a minor myeloid component consisting of 5-15% blast cells with myelomonocytic features, raising the possibility that a diagnosis of bilineal acute leukemia would be more appropriate in such cases. These patients appear to have a severe outcome with significantly lower CR rate than similar cases of Ph-positive ALL without a minor myeloid component.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7496351 TI - CD34+ leukemic cells assessed by different CD34 monoclonal antibodies. AB - CD34 monoclonal antibodies (McAbs) are widely used to identify and isolate hemopoietic progenitors and to classify acute and chronic leukemias. We assessed the reactivity of 17 CD34 McAbs from the 5th International Workshop on Leukocyte Differentiation Antigens with a variety of cells types: normal bone marrow hemopoietic progenitors, 10 AML, 6 ALL, 11 CML. The reactivity for these McAbs was compared with that of reference CD34 McAbs (Q-Bend 10 and 8G12). For each cell population the % of McAb binding cells, the peak channel and the mean fluorescence intensity (MFI) of the positive cells was evaluated. The peak channel, the MFI and the number of positive cells varied significantly from case to case, depending on the McAb and the type of leukemia. According to the spectrum of reactivity three groups of McAbs were defined; however, 7 McAbs do not belong to any of these subgroups. These groups were not entirely in accordance with McAb classification based on enzyme cleavage that identified three epitopes of the CD34 molecule. Some reagents were found to be more specific for AML, other for ALL, CML or normal CD34+ cells. Normal bone marrow light density cells showed a significantly higher percentage of positive cells for 43A1 and MD34.2 McAbs compared to that documented for the remaining McAbs. AML cells showed the most variable pattern of expression for the CD34 McAbs. In leukemic samples, MESF (molecular equivalents of soluble fluorochrome) values ranged from 18,200 to 322,000 and the number of binding sites per cells was 5,000 81,000.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7496352 TI - CD34 expression in adult acute lymphoblastic leukemia. AB - The expression of the pluripotent stem cell antigen CD34 was evaluated at diagnosis in forty-five adult patients with de novo ALL. Comparison of clinical and hematological features between CD34 positive (24/45) and CD34 negative (21/45) patients showed that the former were of older age, had more pronounced lymphoid organ involvement and higher serum LDH levels. Immunophenotypic analysis of marrow blast cells revealed a significant predominance of the 'null' phenotype in the CD34 positive group, together with a strong expression of the VLA-4 and VLA-5 integrins (fibronectin receptors). CD34 positive ALL were also more frequently associated with either aberrant myeloid-related antigens (CD13, CD33) or the P-gp/MDR-1 phenotype. Only 11 out of 24 (45%) CD34 positive patients achieved complete remission after induction chemotherapy, compared to 20/21 (95%) CD34 negative cases. Furthermore, survival was significantly shorter in the CD34 positive group (6.6 mo. vs 13.5 mo.). These results suggest that in ALL, as in AML, CD34 positivity may predict a poor prognosis. PMID- 7496353 TI - Ph positive acute lymphoblastic leukemia in adults: molecular and clinical studies. AB - Fifty-six patients with ALL were investigated for bcr involvement by PCR. Breakpoints were found in 15 patients (26.8%). There were no differences in clinical and hematologic features or the percentages of complete response (CR) between the Ph+ and Ph- cases. The duration of CR was 6 and 8 months, respectively. In 7/9 Ph1 relapsed ALL we observed increased expression of myeloid markers and 2/9 showed a switch of cytotype (Ly-->My). In none of the 13 Ph- relapsed ALL patients did we observe these findings. 7/15 of Ph+ cases expressed P190 and mRNA ela2 and 8/15 patients showed P210, with mRNA b3a2 in 5 and b2a2 in 3, respectively. The percentage of CR was 57% in the P190+ and 87% in the P210+ group. Investigation of more Ph1+ ALL cases treated with a uniform protocol should be performed in the future in order to determine whether any such biological and clinical differences exist. PMID- 7496354 TI - CD34+/CD33+ blast cells: correlation with FAB subtypes. AB - The reproducibility of FAB morpho-cytochemical classification in a multicenter trial was recently assessed as 78.1%. The relevance of immunophenotyping in AML is debated because of the lack of a clear prognostic impact and the necessity of uniform diagnostic criteria and standardised methodologies. Several authors, studying the morpho-immunological characteristics of AML blasts by flow cytometry, suggested the necessity for the definition of clusters of cells with similar morphological patterns and intensity of expression. In this respect we reviewed blast CD34 and CD33 intensity of expression in 65 AMLs classified according to the FAB criteria. Four morphological groups could be identified: 1) low side scatter (SSC) and forward scatter (FSC); 2) intermediate SSC and FSC; 3) high SSC and FSC; 4) combination of pattern 2 plus 3. Five immunological patterns were defined: a) high CD34 expression with negative or weak CD33; b) high CD34 and CD33 expression; c) weak to negative CD34 with high CD33 expression; d) coexistence of 2 different subpopulations; e) negative to weak CD34 and CD33 expression. Based on this morpho-immunological analysis we were able to subdivide AML patients into 5 homogeneous subgroups and a comparison with FAB classification showed a concordance of 73.9%. Regarding CD34 and CD33 intensity of expression, a correlation with prognosis was demonstrated among all M1-M2 patients and in the M2 and M4 subgroups. In conclusion even if immunophenotype cannot substitute the FAB approach to AML, we feel that a flow cytometry morpho immunological analysis could be helpful in achieving a greater understanding and agreement between different Institutions and assist in the definition of more precise prognostic subgroups. PMID- 7496355 TI - Acute myeloid leukemias M2 potentially misdiagnosed as M3 variant French-American Britain (FAB) subtype: a transitional form? AB - From 1990 to 1994, 3 patients with de novo acute myeloid leukemia (AML) in whom light microscopy and cytochemistry suggested a FAB subtype M3 variant were observed at our Institute. Immunophenotype showed HLA-DR-, CD13+, CD33+, CD2+, CD9+; promyelocytic features were also detected by electron microscopy. However, leukemic cells lacked both translocation t(15;17) and RAR alpha/PML genes rearrangement. These cases were considered to be 'M2 atypical' subtypes and they contribute to point out how cytogenetics and molecular biology are mandatory for a correct diagnosis of acute promyelocytic leukemia (APL) particularly because therapy with all trans retinoic acid (ATRA) is now the best treatment for APL. Nevertheless these 3 cases indicate that the atypical M2 subtype may be confused with the M3v if only cytochemistry, immunophenotype and electron microscopy are used in the defining the FAB subtypes. PMID- 7496356 TI - Acute promyelocytic leukemia toluidine blue subtype. AB - In the hypergranular group of acute promyelocytic leukemia (APL) a rare subvariant with basophilic granules, metachromatic for toluidine blue, is recognizable. To evaluate the incidence as well as the biological and clinical significance of this subtype, we studied 53 consecutive untreated patients with APL with morphological, cytochemical, immunological and cytogenetic methods. In 10 cases (19% of the total) granules stained metachromatically in percentages of promyelocytes ranging from 16 to 60. In these cases peroxidase positivity was weaker than in the classic hypergranular and microgranular M3 and activities of esterases were usually present; at the ultrastructural level granules contained particulate material. Immunophenotypic and cytogenetic characteristics seemed not to differ from those of other M3 cases. Coagulopathy was usually life threatening, notwithstanding the low white cell count, and the median survival was short. Hyperhistaminemia-related symptoms were not observed. Cytochemical, immunologic and cytogenetic findings are useful to differentiate this form from M2 with basophilic differentiation and from mast cell leukemia. PMID- 7496357 TI - AML M1 and M2 with eosinophilia and AML M4Eo: diagnostic and clinical aspects. AB - AML with eosinophilia belongs to the morphologic cytogenetic entity M1/M2 t(8;21) with the involvement of the genes AML1/ETO. These eosinophils differ only slightly from normal eosinophils with one rare exception i.e. Auer rods in eosinophils which has only been found on peroxidase staining: This subtype belongs to the good prognosis group of AML. AML with inv(16) (mostly M4Eo) per se is a morphologic-cytogenetic entity with inv(16),--rarely t(16;16), and the genes MYL 11/CBF beta involved. The eosinophils show special abnormalities. AML with inv(16) also belongs to the good prognosis group of AML. PMID- 7496358 TI - Cytophagocytosis by the blast cells in acute myeloid leukemia. AB - The occurrence of cytophagocytosis as spontaneous ingestion of cells by blast cells has been poorly investigated in acute leukemia. Cytophagocytosis by blasts in acute myeloid leukemia (AML) was observed in eleven patients who represent 1% of AML cases seen in our institution (one M1, two M2, four M4 and four M5). Prominent features of cytophagocytosis by AML blasts are particularly evident in leukemias involving monocytic cells and were more obvious in the bone marrow than the peripheral blood. AML of the monocytic type with cytophagocytosis are frequently associated with cytogenetic aberrations involving chromosomes 8 and 16. Finally the poor outcome of AML patients with cytophagocytosis and their poor response to therapy and short survival is stressed. PMID- 7496359 TI - Acute megakaryoblastic leukemia. AB - In 1985 acute megakaryoblastic leukemia was included in the FAB classification system of hematological neoplasias with the designation of AML M7. It occurs in all age groups with two peaks in distribution. The one is in adults and the other in children 1 to 3 years of age especially in those with Down's syndrome. The diagnosis of AML M7 requires more than 30% of the nucleated bone marrow cells being megakaryoblasts. The more common types of AML MO-M6 have to be excluded by morphological and cytochemical analysis whereas immunology is needed to exclude ALL. The megakaryocytic nature of the leukemia has to be proven by ultrastructural demonstration of platelet peroxidase or by immunological demonstration of CD61, CD42, CD41 on the surface of the leukemic blasts. Megakaryocytic/megakaryoblastic leukemias show a wide morphologic spectrum. In some instance small cells dominate, clearly showing megakaryocytic differentiation with scant amounts of cytoplasm and with nuclei showing dense chromatin. On the other hand, there are cases with larger cells resembling ALL-L2 blasts with moderate amounts of rather basophilic cytoplasm which in some instances contain azurophilic granules. Cytoplasmic blebs and protrusions are the most prominent feature of many cases. The nuclei of these cells are round with more finely reticulated chromatin and with prominent nucleoli. The megakaryoblastic nature of these cells can be suggested by morphology. However, according to our experience there are cases of c-ALL with the very same morphologic picture. Consequently, immunologic phenotyping of these cases is necessary in any instance. Cytochemistry is of limited diagnostic value in megakaryoblastic leukemias. Usually it is used to exclude the more common types of leukemia. PMID- 7496360 TI - Techniques for detection of minimal residual disease. AB - Analysis of leukemia-specific and leukemia-associated markers following standard or high-dose treatments is crucial in order to evaluate the efficacy of therapeutic strategies. During the last decade, several techniques have been proposed and used for detecting minimal residual disease (MRD). Each approach is characterized by advantages and limitations, mainly related to its sensitivity and specificity. The general limitations of such tests originates from the size of the sample which can be analysed and the heterogeneous distribution of leukemia after treatment. Clinically useful methods for detecting residual leukemia require not only sensitivity but also speed and reproducibility. The rate of false negative tests is low with polymerase chain reaction as well as flow cytometric analysis. Usually, patients without persistent cells carrying leukemia-associated markers have a lower risk of relapse. However, the detection of a persistent marker at one time point in complete remission cannot be considered a reliable indicator of MRD, whereas increase of positive signals or reappearance of leukemic markers usually precedes relapse. It is likely that one single approach will not allow the monitoring of the majority of patients and that a combination of techniques will be needed. Definitive results will be obtained only through prospective studies in patients receiving standardised therapy. Studies in which therapeutic strategies are designed according to the results provided by techniques for detecting MRD will be necessary to assess the relevance of their contribution to the treatment of leukemia. PMID- 7496361 TI - Acute postobstructive pulmonary edema. AB - Pulmonary edema developing after the relief of upper airway obstruction has been reported in association with a variety of factors including laryngospasm, foreign bodies, and tumors. However, as the phrase "negative pressure pulmonary edema" suggests, markedly negative intrapleural pressure is the dominant mechanism for the genesis of pulmonary edema associated with upper airway obstruction. A review for anesthesia providers of this poorly recognized and often perplexing syndrome may help to reduce the occurrence of this potential complication and facilitate its treatment. PMID- 7496362 TI - Preventing posttraumatic pneumonia. AB - Between 20% and 40% of patients admitted to the hospital with multiple trauma develop bacterial pneumonias. The most immediate causes are aspiration, blunt chest trauma, and prolonged immobilization. Traumatic shock and the need for immediate head and chest surgery are contributing factors. Signs of systemic infection, infiltrates on the chest film, and microbiological evidence obtained from sputum samples are the principal diagnostic criteria. Prophylactic measures include aseptic management of the airway, tracheostomy when indicated, adequate pain relief, and early mobilization. When the incipient signs of pneumonia appear, prompt aggressive antimicrobial therapy is indicated. PMID- 7496363 TI - What's new with reactive airways and anesthesia? PMID- 7496364 TI - Intrathecal narcotics for labor using combined spinal epidural technique: an attractive option for solo anesthesia providers. AB - A 31-year-old primigravida woman presenting with term pregnancy was admitted for labor and delivery. After approximately 10 hours of labor, which included an oxytocin infusion, she requested anesthesia intervention. A combined spinal/epidural (CSE) block was administered featuring a 3 1/2-in, 18-gauge Hustead epidural needle into the epidural space, through which was placed a 4 11/16-in, 27-gauge Whitacre spinal needle into the subarachnoid space. Fifty micrograms of fentanyl, 0.25 mg of morphine, 0.2 mg of epinephrine, and 2 mL of perservative-free normal saline were given intrathecally, the spinal needle was withdrawn, and a catheter placed into the epidural space. Profound analgesia without motor impairment was achieved. After the patient failed to progress, 2% lidocaine was administered epidurally to provide anesthesia to a T4 level, and a cesarean section was performed. The CSE technique provides a safe, flexible choice of anesthesia for labor and delivery. Monitoring of the patient by the anesthetist is necessary for only the initial 30 minutes after block administration. This, coupled with the advantage of having an epidural catheter as a backup, makes it an especially attractive alternative for the solo anesthetist. PMID- 7496365 TI - Ropivacaine: an introduction to a new local anesthetic. AB - Local anesthetic drugs interrupt nerve transmission by inhibiting the passage of sodium ions (Na+) across neuronal membranes. Signal propagation and transmission along central and peripheral nerve pathways are impeded after the injection of these drugs near or on neuronal tissue. This article briefly reviews the basic pharmacology and toxicology of local anesthetic agents as well as recent studies of regional anesthesia performed with ropivacaine, a versatile new amide local anesthetic with a duration of action similar to bupivacaine, but with less systemic toxicity. PMID- 7496367 TI - Anesthetic drug interactions. PMID- 7496368 TI - Post-intubation croup. PMID- 7496366 TI - Optimal monitoring for onset of mivacurium chloride blockade. AB - The purpose of this article is to determine the optimal monitoring for onset of neuromuscular blockade as it relates to mivacurium, a new short-acting nondepolarizing neuromuscular blocker. Clinically, mivacurium has a higher percentage of undesirable consequences to tracheal intubation such as: diaphramatic movement, coughing, and purposeful movement. It is important to understand the time line from onset to recovery with mivacurium and the best muscle to determine onset blockade to minimize these unwanted effects. PMID- 7496369 TI - Synthesis and preliminary evaluation of [11C]KF15372, a selective adenosine A1 antagonist. AB - As a radioligand for mapping the presynaptic adenosine A1 receptors in the central nervous system by PET, [1-propyl-11C]8-dicyclopropylmethyl-1,3 dipropylxanthine ([11C]KF15372), a selective adenosine A1 antagonist, was prepared by the reaction of 8-dicyclopropylmethyl-3-propylxanthine and [11C]propyl iodide with decay-corrected radiochemical yield of 5% based on the [11C]propyl iodide, radiochemical purity of > 99%, sp. at. of 10-56 GBq/mumol and preparation time of 45-55 min. Another 11C-labeled A1 antagonist with much lower affinity for the A1 receptors, 7-[11C]methyl-KF15372 ([11C]KF17109), was also prepared using [11C]methyl iodide with a decay-corrected radiochemical yield of > 50%. In mice, the brain uptake of [11C]KF15372 (1.91% ID/g at 5 min) decreased gradually with time. Carrier KF15372 competitively reduced the brain uptake to a level (43% of the control) comparable to the brain uptake of [11C]KF17109. On the other hand, an A2 antagonist 3,7-dimethyl-1-propargylxanthine showed no effect on the brain uptake of [11C]KF15372. The results show that [11C]KF15372 has potential as a PET radioligand for mapping the adenosine A1 receptors and that [11C]KF17109 may be a reference compound reflecting the non-specific uptake of the [11C]KF15372. PMID- 7496370 TI - Quantitation of reduced disulfide groups in monoclonal antibodies using 5 iodoacetamidofluorescein: a novel size exclusion-HPLC technique. AB - The interference of stannous in the quantitation of sulfhydryl in pretreated MAb using the Ellman's method is described. A HPLC method which effectively overcomes the interference by stannous ion in measuring sulfhydryls by the commonly used Ellman's technique has been developed. The method involves reacting the reduced monoclonal antibody with 0.1 M EDTA followed by the sulfhydryl specific reagent 5 iodoacetamidofluorescein and incubating the mixture at 37 degrees C for 1.0-1.5 h. The 5-IAF labeled protein is separated from the unbound free 5-IAF and its secondary degraded products by size exclusion HPLC using a TSK G3000 SWXL column. The method allows quantitation of MAb sulfhydryl present as monomeric MAb, aggregates and fragments and is a major improvement over the spectroscopic techniques. PMID- 7496371 TI - Instantaneous preparation of radiolabeled 5-iodo-2'-deoxyuridine. AB - The fastest and most reliable preparation of radiolabeled 5-iodo-2'-deoxyuridine ([123/125/131I]IUdR) is accomplished by iododemetallation. We describe a series of improvements to a previously published synthesis of radiolabeled IUdR by demercuration whereby [123/125/131I]IUdR is synthesized instantly following the incubation of an aqueous solution of the chloromercury precursor with Na123/125/131I in the presence of Iodogen. The total procedure is completed in 5 min and does not require purification. Mercury content is low and HPLC indicates the total transformation of iodide into IUdR with no detectable UV-absorbing by products. PMID- 7496373 TI - Radiation dosage accumulated by reindeer from Novaya Zemlya. AB - ESR spectrometry of tooth enamel revealed a significant difference in accumulated radiation dose between reindeer which lived on Novaya Zemlya when underground nuclear tests were performed there and those which lived there after the tests were stopped. PMID- 7496372 TI - Factors determining the radioactivity levels of waters in the province of Caceres (Spain). AB - In an extensive study of different types of water in the province of Caceres (Spain) in order to determine their total alpha- and total beta-radioactivity contents, several factors were found to have a decisive influence on their radioactivity levels. Thus, parameters such as total hardness and pH can be determinative according to the lithological type of the aquifer or according to the subterranean or surface origin of the hydrological resource. Waters from classic lithologies, or originating from a well, possess above-average radioactivity indices, surpassing in numerous cases the indicative levels set by current Spanish legislation. In contrast, surface waters from rivers, in practically all cases, possess below-average radioactivity indices. PMID- 7496375 TI - Influence of the administered mass of tellurium on plasma clearance in rabbits. AB - The combination of analytical techniques such as PNA and SIMS with a compartmental approach enables the study of the metabolism and biokinetics in humans of several elements by using stable isotopes as tracers. The techniques developed for Te require the administration of greater masses than those used for similar studies performed with radioactive tracers, therefore a test was carried out in rabbits in order to assess the possible influence of the administered amounts on the determination of the biokinetic parameters. The behaviour of the tracers was found to be similar for Te administration of up to 70 micrograms/kg of body weight. An inter-individual variability in the size of the transfer compartment was observed and has to be taken into account. PMID- 7496374 TI - [125I]IgM (KAU) human monoclonal cold agglutinin: labelling and studies on its biological activity. AB - In order to study the interaction between an IgM cold agglutinin and the erythrocyte I antigen, the former antibody was labelled with 125I using the Chloramine-T, IODOGEN and Bolton-Hunter methods. High incorporation and adequate stability of the labelled IgM were obtained with all procedures. However, suitable biological activity was maintained only with the Bolton-Hunter method. Further studies suggest that tyrosine iodination affects antigen recognition by this IgM, whereas iodination of amino groups does not. The reagent thus prepared allowed the determination of the number of I sites per erythrocyte as well as the antibody affinity constant. PMID- 7496376 TI - Solute sensing vs. solvent sensing, a speculation. PMID- 7496377 TI - The regulation of Dictyostelium development by transmembrane signalling. AB - Dictyostelium discoideum has a well characterized life cycle where unicellular growth and multicellular development are separated events. Development is dependent upon signal transduction mediated by cell surface, cAMP receptor/G protein linkages. Secreted cAMP acts extracellularly as a primary signal and chemoattractant. There are 4 genes for the distinct cAMP receptor subtypes, CAR1, CAR2, CAR3 and CAR4. These subtypes are expressed with temporally and spatially specific patterns and cells carrying null mutations for each gene have distinct developmental phenotypes. These results indicate an essential role for cAMP signalling throughout Dictyostelium development to regulate such diverse pathways as cell motility, aggregation (multicellularity), cytodifferentiation, pattern formation and cell type-specific gene expression. PMID- 7496378 TI - A novel opioid mechanism seems to modulate phagocytosis in Tetrahymena. AB - We have previously reported that a beta-endorphin-like substance inhibits phagocytosis in Tetrahymena perhaps by a mu-like opioid receptor. We now report a further characterization of the elements involved in the signal transduction mechanism of this opioid. Affinity chromatography followed by immunoblots of both intracellular extracts and extracellular medium reveal the presence of two main proteins of 64 and 75 kDa. These molecular weights are much higher than that of any known opioid peptide or precursor protein and suggest that we may be dealing with either a novel opioid or with proteins that by chance cross-react with anti beta-endorphin antibody. Nevertheless, when the biological activity of these proteins was tested it was found that they had an effect similar to that of mammalian beta-endorphin, namely inhibition of phagocytosis by a naloxone reversible mechanism. We have probed a size-selected Tetrahymena library with a pro-opiomelanocortin probe and have obtained several positive clones; the sequencing of their inserts should establish whether we are dealing with a bona fide member of the opioid family. Another aspect we have been studying is the G proteins which appear to be involved in the modulation of phagocytosis. We have found, by means of Western blotting (using an antibody against the conserved GTP binding region of the alpha-subunit), two bands of 51 and 59 kDa; no alpha subunit of 59 kDa had been reported previously and may represent a novel G protein. In spite of these differences, the opioid signal transduction mechanism appears to remarkably resemble that present in more complex organisms. PMID- 7496379 TI - Chemical signaling in ciliates. AB - For long, our knowledge of the biology of ciliate pheromones has long relied solely upon the study of the two structurally unrelated "gamones" identified in culture filtrates of a Blepharisma species. However, the characterization of a number of polypeptide pheromones secreted by Euplotes raikovi and E. octocarinatus has now established that structural relationships of homology usually link these molecules, which is consistent with the genetic basis of the mating type systems evolved by these species. In this context, our growing appreciation of the conserved and variable elements of the pheromone architecture should foster progress in the understanding of pheromone-receptor interactions and thus, provide important clues into pheromone mechanisms of action. PMID- 7496380 TI - Effects of temperature on promastigotes of several species of Leishmania. AB - Six Leishmania species were studied comparatively, in order to determine the influence of temperature "in vitro" on differentiation, infectivity and protein synthesis. Differentiation occurred in a heterogeneous manner, even in species that produce similar clinical manifestations. Thus, no association could be found between thermosensitivity and disease. The association between expression of proteins and increasing temperatures was analyzed at 34 degrees C by polyacrylamide gel electrophoresis with sodium dodecyl sulphate (SDS-PAGE), using different incubation times, and employing a technique involving metabolic incorporation of [35S]-methionine. Protein synthesis was very similar in all the New World species apart from L. amazonensis, which expressed a protein of approximately 80 kDa when incubated at 34 degrees C for 2 hours. All the tested species had in common the expression of a 70 kDa protein. Differences, however, were observed in relation to the time interval for protein expression. In L. chagasi, synthesis was detected after 30 minutes of incubation at 34 degrees C, while L. braziliensis required 1 hour at the same temperature. The "in vivo" and "in vitro" infectivity of the differentiated forms was also analyzed, but no significant differences were observed. PMID- 7496381 TI - Plasmodium falciparum rhoptry proteins of 140/130/110 kd (Rhop-H) are located in an electron lucent compartment in the neck of the rhoptries. AB - To investigate in more detail the structure of the high molecular weight rhoptry protein complex of Plasmodium falciparum, Rhop-H (140/130/110 kd), the complex was affinity purified from parasite extracts using rhoptry protein specific antisera prepared against Rhop-H proteins bound to and eluted from Balb/c mouse erythrocytes, using 0.5 M NaCl. The individual proteins (140 kd/Rhop-1, 130 kd/Rhop-2, and 110 kd/Rhop-3) were separated, electroeluted, and monospecific polyclonal antisera prepared against the individual proteins, and against the affinity purified complex. Immunofluorescence assays and immunoelectron microscopic studies were performed to verify the subcellular localization of the Rhop-H epitopes. Immunoblotting and immunoprecipitation assays were also performed. We report novel findings regarding the localization of the rhoptry proteins to an electron lucent compartment in the neck of the rhoptries. Analysis of the amino acid composition of the individually purified Rhop-H proteins demonstrated a predominance of negatively charged (E, D) as well as hydrophobic residues (L, A, P, S) in the three proteins. The percentage of negatively charged residues was high for all three proteins. Similarities in amino acid composition for the three proteins supports the previous data demonstrating shared properties such as erythrocyte and liposome binding, for the three proteins. Results of antibody characterizations using rhoptry protein specific antisera demonstrate the immunodominance of the Rhop-H complex. PMID- 7496382 TI - Isolation of N-acetyl-beta-hexosaminidase from Acanthamoeba castellanii. AB - The lysosomal enzyme N-acetyl-beta-hexosaminidase (beta hex) has been purified from Acanthamoeba castellanii growth medium by a three step procedure. The enzyme was precipitated with ammonium sulfate, partially purified on a DE52 column and purified to homogeneity on an affinity column. The purified beta hex appeared to be a monomer with a molecular mass of 58 kDa and a pI of approximately 5.8. The enzyme activity in growth medium at RT was stable for several months. The purified beta hex was enzymatically deglycosylated and injected into two rabbits to make polyclonal antibodies. One antiserum was specific for beta hex, but the other stained many bands on immunoblots of whole cell preparations. Using fluorescently labelled secondary antibodies we have determined that both antisera stain digestive vacuoles in the Acanthamoeba cytoplasm, and do not stain the contractile vacuole. The multi-specific antiserum had high avidity for beta hex, but also stained the carbohydrate portion of other molecules. These other molecules may be lysosomal enzymes as well, since the activity of several other lysosomal enzymes was partially immunoprecipitable with the antiserum. We plan to use these antibodies to study traffic patterns among the variety of vacuolar structures in Acanthamoeba cytoplasm. PMID- 7496383 TI - pH-dependent effects of 2,4-dinitrophenol (DNP) on proliferation, endocytosis, fine structure and DNP resistance in Tetrahymena. AB - In view of the importance of external pH on cytotoxic effects of ionizable agents, the pH-dependent effects of 2,4-dinitrophenol (DNP) were investigated. As uncoupler of oxidative phosphorylation, DNP interferes with the proton gradient across the inner mitochondrial membrane. DNP was added to proliferating Tetrahymena pyriformis in media of different initial pH. Effects studied were rates of cell proliferation and endocytosis, and fine structure. Findings correlated with the calculated concentration of undissociated DNP, taking into account that pH changes with time and cell density in Tetrahymena cultures. A linear relationship thus emerged between initial concentrations of undissociated DNP and lengths of the lag preceding cell proliferation. Once resumed, the rate of proliferation corresponded to that of control cells, even in different concentrations of undissociated DNP, presumably indicating an adaptation mechanism. Endocytosis was elevated throughout a wide range of undissociated DNP concentrations with a sharp transition towards inhibition at high DNP concentrations causing lethality with time. Changes in fine structure of DNP treated cells (mitochondria, peroxisomes, nucleoli) also depended on the concentration of undissociated DNP. PMID- 7496384 TI - The NADP-linked aldehyde reductase of Leishmania donovani. AB - An enzyme that oxidizes ethanol to acetaldehyde in the presence of NADP (but not NAD) and reduces acetaldehyde to ethanol in the presence of NADPH (but not NADH) is present in Leishmania donovani promastigotes. The activity is present only in the supernatant fraction obtained from sonication of the cells and high speed centrifugation. The Km and Vm values were evaluated for propanol and propionaldehyde as well as for ethanol and acetaldehyde in cells obtained from late log and 3-day stationary phase cultures. There was no significant change in Km or Vm values for any of these four substrates with culture age. Since the Km values for ethanol and propanol are much higher than for the corresponding aldehydes and higher than any physiological range of alcohol concentration likely to be encountered, this enzyme is considered to function as an aldehyde reductase. PMID- 7496385 TI - YI-S, a casein-free medium for axenic cultivation of Entamoeba histolytica, related Entamoeba, Giardia intestinalis and Trichomonas vaginalis. AB - Pancreatic digests of casein are major ingredients of media used in the axenic cultivation of lumen-dwelling parasitic protozoa, especially Entamoeba, Giardia, and trichomonads. The digest used almost exclusively in the development of these media, Medo-Peptone (Trypticase BBL), has not been available since 1981. Moreover, none of dozens of similar type digests tested since then in our laboratory has proved equal to Medo-Peptone, and in the last two years it has become increasingly difficult to obtain new batches which will support even modest growth of Entamoeba histolytica. In response to this problem we have developed a casein-free medium, YI-S, consisting of a nutrient broth, vitamin mixture and serum. We recommend it as a replacement for the casein-dependent medium TYI-S-33, currently the most widely used for axenic culture of Entamoeba histolytica and other lumen-dwellers. PMID- 7496386 TI - Long-range restriction mapping of megabase-sized chromosomes that may be homologs in Trypanosoma brucei. AB - Trypanosoma brucei is a blood-borne pathogen that changes its variant surface glycoprotein coat, thus evading immune destruction. Restriction digestion, combined with probe hybridization studies, was used to construct long-range restriction maps of the 1.4 (M4) and 1.5 megabase (M3) chromosomes from the IsTaR1 serodeme of T. b. brucei. Comparison of the two chromosomes suggests that they are a homologous pair. Hybridization with a repetitive sequence probe also identifies several copies on the M4 chromosome and a relative paucity of cross hybridizing repetitive sequence on the larger M3 chromosome. PMID- 7496387 TI - Inhibition of protein synthesis and amino acid transport by crystal violet in Trypanosoma cruzi. AB - [35S]methionine incorporation into proteins of either T. cruzi epimastigotes or trypomastigotes was drastically inhibited by low concentrations of crystal violet in a dose-dependent manner. This inhibition was not due to ATP depletion since cellular ATP levels did not change significantly after incubation of epimastigotes with 50 microM crystal violet for similar periods of time, and was unaffected by changes in the extracellular free calcium concentration. Although crystal violet was able to inhibit protein synthesis in a cell-free system from T. cruzi epimastigotes, half maximal inhibition was at 1 mM, a concentration three orders of magnitude higher than those that inhibited protein synthesis in intact cells. On the other hand, crystal violet was able to inhibit total [35S]methionine uptake at similar concentrations to those that inhibited protein synthesis while addition of increasing concentrations of cold methionine to the incubation medium protected the cells against crystal violet inhibition. Crystal violet also inhibited total [3H]proline uptake thus indicating that it has a general inhibitory effect upon the transport of amino acids, and not specifically upon methionine. These results indicate that inhibition of protein synthesis by crystal violet is probably due to inhibition of amino acid uptake. PMID- 7496389 TI - Effect of fetal bovine serum glycoproteins on the in vitro proliferation of the oyster parasite Perkinsus marinus: development of a fully defined medium. AB - The oyster parasite Perkinsus marinus replicates in our medium consisting of Dulbecco modified Eagle's medium: Ham's F12 nutrient mixture (1:1) supplemented with 1-5% fetal bovine serum, with a doubling time of 24 hours during the exponential phase of the culture. Fetal bovine serum concentrations above 5% dramatically reduced parasite proliferation in a dose-dependent manner. We tested the individual effects of the three major protein components of fetal bovine serum (fetuin, transferrin and albumin) on the replication of the parasite in a serum-free medium. At the concentrations tested, fetuin enhanced parasite growth, whereas albumin had a modest positive effect and transferrin was inhibitory. Proteolytic digestion of fetuin, strongly diminished its growth-enhancing properties, indicating that the overall glycoprotein architecture may be required for activity. On the contrary, desialylation of fetuin slightly enhanced its growth-promoting activity. The addition of fetuin at 1.7 mg/ml to the serum-free DME:Ham's F12 medium yielded growth rates that are comparable to those obtained with our standard culture methodology. This has resulted in a fully defined culture medium that will allow for a rigorous characterization of excretory/secretory products involved in modulating or blocking the host's humoral and cellular defense mechanisms. PMID- 7496390 TI - Comparison of the 24 kDa flagellar calcium-binding protein cDNA of two strains of Trypanosoma cruzi. AB - DNA sequences encoding the 24 kDa flagellar calcium binding protein (FCaBP) of two strains of Trypanosoma cruzi were found to differ at fourteen positions, six of which result in amino acid differences. Four of the amino acid differences are located within the calcium-binding domains of FCaBP; however, none is predicted to affect the calcium-binding ability of the protein. Chromosomes harboring the FCaBP gene clusters differ in size among T. cruzi strains. PMID- 7496388 TI - Cloning and characterization of an ATPase gene from Pneumocystis carinii which closely resembles fungal H+ ATPases. AB - A gene encoding a P-type cation translocating ATPase was cloned from a genomic library of rat-derived Pneumocystis carinii. The nucleotide sequence of the gene contains a 2781 base-pair open reading frame that is predicted to encode a 101,401 dalton protein composed of 927 amino acids. The P. carinii ATPase protein (pcal) is 69-75% identical when compared with eight proton pumps from six fungal species. The Pneumocystis ATPase is less than 34% identical to ATPase proteins from protozoans, vertebrates or the Ca++ ATPases of yeast. The P. carinii ATPase contains 115 of 121 residues previously identified as characteristic of H+ ATPases. Alignment of the Pneumocystis and fungal proton pumps reveals five homologous domains specific for fungal H+ ATPases. PMID- 7496391 TI - Antisense oligonucleotides in the study of neuroendocrine systems. PMID- 7496392 TI - Neuronal activity and neuropeptide gene transcription in the brains of immune challenged rats. AB - The present study investigated the effect of the acute-phase response of a systemic immune activation on the transcription of various immediate early genes (IEGs) and neuropeptides in the brain of conscious rats. One, 3, 6, 9, and 12 h after a single intraperitoneal (i.p.) administration of either the immune activator lipopolysaccharide (LPS) or the vehicle solution, adult male rats were sacrificed and their brains cut in 30-microns coronal sections. mRNA encoding the IEGs c-fos and nerve growth factor inducible-B (NGFI-B), and neuropeptides corticotropin-releasing factor (CRF), oxytocin (OT), and vasopressin (AVP) were assayed by in situ hybridization histochemistry using a 35S-labeled riboprobes. The primary transcripts [heteronuclear (hn)RNA] for these neuropeptides were also detected using intronic probe technology, and colocalization of c-fos mRNA within CRF, AVP, and OT neurons was determined by means of a combination of immunocytochemistry and in situ hybridization techniques on same the brain sections. One h after LPS treatment, both c-fos and NGFI-B genes were expressed in the parvocellular division of the paraventricular nucleus (PVN) of the hypothalamus. The medial preoptic area/organum vasculosum of the lamina terminalis, the supraoptic nucleus (SON), the magnocellular division of the PVN, the arcurate nucleus/median eminence, the locus coeruleus, the nucleus of the solitary tract, and the area postrema also exhibited a strong signal for these two transcripts 3 h after endotoxin administration. A smaller but a significant c fos expression was observed in various structures, including the dorsomedial hypothalamic area, the central nucleus of the amygdala, the ventral part of the tuberomammillary nucleus, the laterodorsal tegmental nucleus, the external lateral part of the parabrachial nucleus, the dorsal division of the ambiguus nucleus, and the lateral reticular nucleus of LPS-injected rats. The signal for c fos and NGFI-B mRNA in most of these brain nuclei reached a maximum at 3 h postinjection, declined at 6 h, and vanished 9 to 12 h after LPS treatment. In the parvocellular nucleus of the PVN, c-fos was largely expressed in CRF immunoreactive (ir) neurons, whereas in the magnocellular part of that nucleus and in the SON, this transcript was colocalized in numerous OT-ir and few AVP-ir neurons. Relative levels of CRF mRNA in the parvocellular PVN were also significantly increased 6 h following LPS, but endotoxin did not alter the genetic expression of this stress-related neuropeptide in other brain regions.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 7496394 TI - Regulation of melatonin secretion by light in the isolated pineal organ of the white sucker (Catostomus commersoni). AB - The effects of different lighting conditions and physical parameters of light were investigated in the isolated pineal organ of the white sucker kept under static or superfusion culture. The secretion of pineal melatonin is directly controlled by the photoperiod and completely suppressed under constant illumination. When pineal organs are exposed to unexpected light at night, the secretion of melatonin is significantly reduced within 15 min and reaches basal value after 30-35 min of light treatment. The inhibition of melatonin secretion by unexpected light at night depends on the irradiance, duration, timing of the treatment and the lighting history of the pineal organ. PMID- 7496393 TI - Differential melanin-concentrating hormone gene expression in two hypothalamic nuclei of the teleost tilapia in response to environmental changes. AB - For some teleosts, a role has been established for melanin-concentrating hormone (MCH) background adaptation and stress response. In teleost fishes, prepro-MCH (ppMCH) mRNA is expressed in the hypothalamus, predominantly in neurons of the nucleus lateralis tuberis (NLT) and in scattered cells of the nucleus recessus lateralis (NRL). The response of mature tilapia to different environmental challenges was studied by assessing ppMCH mRNA levels in these two hypothalamic nuclei by quantitative dot blot analysis. Changes in background colour induced pronounced differences in ppMCH mRNA expression in the NLT, but not in the NRL. The NLT of tilapia adapted to a white background contained 2.5 to 3 times more ppMCH mRNA than the NLT of black-adapted fish. The NLT of fish kept on neutral background contained intermediate levels of ppMCH mRNA, which were significantly lower than the levels in white-adapted fish. Oral administration of dexamethasone lowered plasma cortisol concentrations, but had no effect on ppMCH mRNA levels in white- and black-adapted fish. In tilapia exposed to strongly acidified water (pH 3.5), plasma cortisol and ACThH concentrations were highly elevated, and plasma chloride concentrations considerably lower than in controls. These fish responded with a 70% rise in ppMCH mRNA levels in the NLT, which is most probably associated with a stress response evoked by inadequate osmoregulation. After exposure to a milder acidification (pH 4.0) or to seawater no significant changes in ppMCH mRNA levels occurred in either the NLT or the NRL, nor in plasma chloride, cortisol and ACTH levels. A specific increase of ppMCH mRNA levels in the NRL was observed in repeatedly disturbed tilapia.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7496395 TI - Sexually dimorphic expression of androgen receptor immunoreactivity by somatostatin neurones in rat hypothalamic periventricular nucleus and bed nucleus of the stria terminalis. AB - Gonadal steroids exert important regulatory actions on the hypothalamic neurones regulating growth hormone secretion and are believed to play a role in generating its sexually dimorphic pattern of secretion. Recent evidence indicates that estrogen actions on one of these neural populations, the periventricular somatostatin (SOM) neurones, are likely to be indirect as they do not possess nuclear estrogen receptors in either sex although androgen receptors (ARs) have been reported within these cells in male rats. The present study has used double labelling immunocytochemistry procedures to examine whether sex differences exist in AR expression by SOM neurones located in the periventricular nucleus and bed nucleus of the stria terminalis (BNST). Within the hypothalamus, SOM immunoreactive neurones were found concentrated in the periventricular nucleus while both anterior and posterior divisions of the BNST contained scattered populations of SOM cells. Cells immunoreactive for the AR were detected in all of these areas. Although the intensity of AR cell nuclei staining was equivalent in males and females in regions such as the lateral septum, the intensity of AR staining in many individual cells of the periventricular nucleus and posterior BNST of the female was reduced when compared with the male. Double-labelling experiments revealed that approximately 40% of periventricular SOM neurones expressed AR immunoreactivity in the male compared with significantly (P < 0.01) fewer cells in the female (approximately 7%). In the BNST, double-labelled cells were only detected within the principle encapsulated, interfascicular and transverse nuclei of its posterior division. Approximately 60% of SOM cells in these nuclei expressed AR immunoreactivity in the male while significantly (P < 0.01) fewer did so in the female (approximately 25%). These results indicate that substantial sex differences exist in AR expression by SOM neurones in both the periventricular nucleus and BNST. Such differences in AR expression by periventricular SOM cells may contribute to their sexuality dimorphic nature and, consequently, sex differences in growth hormone secretion. PMID- 7496396 TI - Effects of N-methyl-D-aspartate (NMDA) on seasonal cycles of reproduction, body weight and pelage colour in the male Siberian hamster. AB - Siberian hamsters (Phodopus sungorus) transferred from stimulatory photoperiods (long days: LD) to inhibitory photoperiods (short days: SD) undergo testicular regression within 8 weeks. This reproductive response to photoperiod was blocked by systemic daily treatment with the glutamatergic agonist N-methyl-D-aspartate (NMDA: 20 mg/kg BW, sc). This powerful effect of NMDA demonstrates the potential for endogenous glutamate to regulate reproductive function. The overall aim of the subsequent studies was to investigate the site and mechanism of action of this glutamatergic agonist in order to identify potential mechanisms through which endogenous glutamate might act. To investigate whether the effect of systemic NMDA was via an effect on the circadian timing system, alterations in gonadal regression and recrudescence, seasonal coat changes (pelage) and body weight (BW) were examined. It would be predicted that long-term cycles of all these seasonal parameters would be affected if the action of NMDA were to perturb the transduction of photoperiodic information. Daily treatments with NMDA, which initially maintained reproductive function in hamsters exposed to SD, did not influence the time course of subsequent testicular recrudescence, nor did they influence long-term cycles of pelage and BW. Moreover, treatment with NMDA induced a dose-dependent increase in serum concentrations of LH within 15 min of systemic injection. These data are consistent with the hypothesis that systemic NMDA exerts it reproductive effects not via an action on the circadian system, but via an action on secretion of GnRH. To investigate potential central sites of action of glutamate, induction of the immediate early gene c-fos, an acute marker of cellular response, was evaluated immunocytochemically (ICC) in brain areas after treatment with NMDA. Although dual-label ICC studies revealed that NMDA did not induce c-fos within GnRH neurons, NMDA did induce c-fos in many cells in the region of the organum vasculosum of the lamina terminalis (OVLT), an area containing a large number of GnRH perikarya, and in the arcuate nucleus, a region close to GnRH secretory terminals in the median eminence. The lack of c-fos induction of GnRH cells argues against a direct effect of NMDA on GnRH neurons. Thus, we examined immunocytochemically the distribution of the common NMDAR1 glutamate receptor subunit to evaluate further the potential sites of glutamatergic action. As expected, NMDAR1-ir was widespread in perikarya throughout the brain, including the region of the OVLT and the arcuate nucleus.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 7496397 TI - Species differences in estrogen receptor and progesterone receptor-mRNA expression in the brain of sexual and unisexual whiptail lizards. AB - Circulating concentrations of gonadal steroid hormones and reproductive behavior in female vertebrates vary as a function of ovarian state. Steroids secreted by the ovary, specifically estrogen and progesterone, influence the expression of behaviors associated with reproduction by intracellular sex steroid receptors located in specific regions of the brain. Using in situ hybridization, we analyzed estrogen receptor and progesterone receptor messenger RNA expression in several brain regions of ovariectomized, vitellogenic, and postovulatory individuals from two species of whiptail lizards (Cnemidophorus uniparens and C. inornatus). Although these species are genetically very similar, they differ in two aspects of their reproductive biology: (i) the unisexual C. uniparens alternate between expressing female-typical and male-like pseudosexual behaviors while female C. inornatus normally express only female receptive behavior, and (ii) circulating estradiol concentrations in reproductively active female C. uniparens are approximately five-fold lower than in reproductively active female C. inornatus. We found that the regulation of sex steroid receptor gene expression was region specific, with receptor-mRNA expression being increased, unchanged, or decreased during vitellogenesis depending on the area. Furthermore, several species differences in the amount of sex steroid receptor-mRNA were found that may be relevant to the species differences in circulating estrogen concentrations and sexual behavior. PMID- 7496398 TI - Subunit E of the vacuolar H(+)-ATPase of Hordeum vulgare L.: cDNA cloning, expression and immunological analysis. AB - A tonoplast protein of 31 kDa apparent molecular mass (TpP 31) was isolated from two-dimensional gels. Amino acid sequences were determined from LysC endoproteinase-peptide fragments. Using degenerate oligonucleotides, a corresponding cDNA clone of 1034 bp was isolated from a barley leaf cDNA library. It encodes for subunit E of the vacuolar H(+)-ATPase, the first one identified in plants so far. The open reading frame extends over 681 bp, encoding a gene product of 227 amino acids and a calculated molecular weight of 26,228 g mol-1. Northern and Western blot analysis indicates constitutive expression of subunit E in all plant organs with only small effects of salt stress. Localization of TpP 31 at the tonoplast was confirmed in fractions of purified vacuolar membrane obtained by free-flow electrophoresis. Immunoprecipitation of newly synthesized 35S-labelled membrane proteins with anti-TpP 31 gave two additional bands with apparent molecular masses of about 53 and 62 kDa. Gel filtration after mild solubilization showed co-purification of TpP 31 with the 55 kDa subunit of the H(+)-ATPase. Both results provide evidence beyond the sequence homology that TpP 31 is a structural component of the vacuolar H(+)-ATPase. PMID- 7496399 TI - The distribution of a spliceosome protein in cereal (Triticeae) interphase nuclei from cells with different metabolic activities and through the cell cycle. AB - A monoclonal antibody (mAb) KSm2 and three human Sm sera, which detect the 'D' polypeptide in mammals and is associated with the U1, U2, U4/U6, U5, U7, U9-U12 small nuclear RNAs, have been used in Western blotting to show that the antigen is conserved in wheat (Triticum aestivum cv. Beaver). Immunocytochemistry to wheat and a barley (Hordeum vulgare) suspension culture using the mAb KSm2 has shown that the 'D' polypeptide occurs in interphase nuclei as speckles and foci outside the nucleoli and as tracks. Nucleoli usually had at least one focus at their periphery. Immunocytochemistry at the electron microscope level of resolution has shown that the signal occurs between chromatin axes and predominantly in the outer domain of the nucleus. Analysis of the barley suspension culture demonstrated that there was a significant increase in antigen through G1, S to G2. In the wheat meristematic cells at prophase only the foci remained, and at metaphase the distribution of foci was asymmetrical with foci occurring either at one pole or on the metaphase plate. At telophase the foci appeared to decrease in size and were incorporated non-uniformly into the daughter nuclei because of their asymmetric distribution at metaphase. When wheat was grown at a range of temperatures, the root tip meristematic nuclei showed a number of different organization patterns. The average number of nucleoli increased and their size decreased. At the same time there was an increase in the average total area of foci as temperature increased from 4 degrees, 10 degrees, 25 degrees, 34 degrees, to 37 degrees C. Thus the 'D' polypeptide distribution changes with metabolic activity and through the cell cycle. PMID- 7496400 TI - The use of an alternative promoter in the Arabidopsis thaliana HMG1 gene generates an mRNA that encodes a novel 3-hydroxy-3-methylglutaryl coenzyme A reductase isoform with an extended N-terminal region. AB - The enzyme 3-hydroxy-3-methylglutaryl-CoA reductase (HMGR, EC 1.1.1.34) catalyses the synthesis of mevalonate, the committed precursor of the great variety of isoprenoid compounds and derivatives synthesized in higher plants. It has previously been reported that Arabidopsis thaliana contains two differentially expressed genes, HMG1 and HMG2, that encode two HMGR isoforms (HMGR1 and HMGR2, respectively). This paper reports the characterization of a novel HMGR mRNA (HMGR1L mRNA) derived from the HMG1 gene. This mRNA is initiated 121 bp upstream from the transcription start site previously characterized. In contrast with the previously reported HMGR1 mRNA (HMGR1S mRNA), which is detected at high levels in all tissues of the plant, HMGR1L mRNA is present at relatively low levels and its expression is restricted mostly to seedlings, roots and inflorescences. HMGR1L and HMGR1S mRNAs are transcribed from alternative promoters. HMGR1L mRNA contains an in-phase AUG start codon which allows the synthesis of a novel HMGR isoform (HMGR1L) having 50 additional amino acid residues at its N-terminal end. Using an in vitro transcription-translation system we have shown that HMGR1L is inserted into ER-derived microsomes. It is thus unlikely that the extended N-terminal region of HMGR1L might have a role in targeting the enzyme to plastids or mitochondria. These results support the previous proposal that the endoplasmic reticulum is the only cell compartment for the primary targeting of HMGR in Arabidopsis and reinforce the view that plant HMGR is under the control of complex mechanisms operating at both transcriptional and post-transcriptional levels. PMID- 7496401 TI - A new elicitor of the hypersensitive response in tobacco: a fungal glycoprotein elicits cell death, expression of defence genes, production of salicylic acid, and induction of systemic acquired resistance. AB - A 32 kDa glycoprotein whose effects in tobacco and other Nicotianae mimic a typical hypersensitive response, was isolated from Phytophthora megasperma. Infiltration of a few nanograms of the protein into leaves caused the formation of lesions that closely resemble hypersensitive response lesions. Transcripts of genes encoding enzymes of the phenylpropanoid and sesquiterpenoid pathways accumulated rapidly after elicitor application followed by salicylic acid production. Cellular damage, restricted to the infiltrated zone, occurred only several hours later, at a time when expression of PR protein genes was activated. After several days systemic acquired resistance was also induced. Thus, tobacco plant cells that perceived the glycoprotein generated a cascade of signals acting at local, short, and long distances, and causing the coordinate expression of specific defence responses in a way similar to hypersensitivity to tobacco mosaic virus. The glycoprotein represents a powerful tool to investigate further the signals and their transduction pathways involved in induced disease resistance. It may also be useful to engineer broad disease protection in a Nicotianae and possibly into crop plant species. PMID- 7496402 TI - Maternal-specific demethylation and expression of specific alleles of zein genes in the endosperm of Zea mays L. AB - Zeins constitute 60-70% of maize endosperm protein. Zein genes are specifically transcribed in the endosperm, and a correlation has been established between tissue-specific expression and demethylation. Three inbred lines and their reciprocal crosses were analysed to assess for allele-specific differences in methylation, transcription and translation. DNAs from endosperm, embryo and seedling tissues analysed by cleavage with methylation-sensitive restriction enzymes and Southern blot hybridization with zein cDNA and genomic sequences show that specific demethylation of zein sequences occurs only in endosperm and is restricted to the maternal complements. Steady-state transcript accumulation of zein mRNA assessed by RNase protection assay reveals qualitative and quantitative differences among endosperm RNAs of the inbreds and of their reciprocal hybrids. Moreover, two-dimensional gel electrophoresis of zein proteins identified polypeptides that are maternally imprinted in reciprocal crosses. These results indicate that endosperm-specific expression of specific zein alleles may occur via parental imprinting and disclose a possible role of methylation in regulating the expression of genes differently contributed in the endosperm by the maternal and paternal genomes. PMID- 7496404 TI - Derepression of the activity of genetically engineered heat shock factor causes constitutive synthesis of heat shock proteins and increased thermotolerance in transgenic Arabidopsis. AB - ATHSF1 is a heat shock transcription factor (HSF) of Arabidopsis that is constitutively expressed but its activity for DNA binding, trimer formation and transcriptional activation of heat shock (hs) genes is repressed at normal temperatures. In this study the functional properties of chimeric HSF glucuronidase (GUS) fusion proteins were tested. Ectopic expression of HSF-GUS or GUS-HSF in transgenic Arabidopsis plants resulted in a derepression of HSF functions as shown by trimer formation, specific DNA binding, and the constitutive expression of heat shock proteins (HSPs) at normal temperature. A novel GUS activity-staining protocol was used to show the specific binding of trimeric HSF fusion proteins to DNA and following hs, an interaction between chimeric HSF-GUS and authentic HSF proteins. The chimeric HSFs were insensitive to the negative regulation that counteracts activation of the authentic HSF at normal temperature. Heterotrimer complexes were reconstituted in vitro from recombinant ATHSF1 and HSF-GUS proteins expressed in Escherichia coli and using this protocol, the temperature-dependent activation of wt HSF was monitored in vivo and in vitro. Transgenic plants expressing constitutively active HSF-GUS fusion proteins are also constitutive for HSPs. Approximately 20% of the maximum heat-inducible levels of HSP18 were already present at normal temperature. The level of basic thermotolerance was significantly enhanced in these plants. The results indicate that genetic engineering using protein fusion is a very effective means to derepress the activity of an important regulatory protein in plants, that consequently activates a constitutive hs response in the absence of heat stress and eventually alters the thermotolerance phenotype. PMID- 7496403 TI - Immunolocalization of freezing-tolerance-associated proteins in the cytoplasm and nucleoplasm of wheat crown tissues. AB - A protein family associated with the development of freezing tolerance in wheat has been identified. This protein family is Gramineae-specific and coordinately regulated by low temperature. Antibodies directed against the 50 kDa (WCS120) protein recognize at least 5 members of this family. Using these antibodies, the cellular content and location of this protein family was determined in cold acclimated wheat seedlings. Western analyses of subcellular fractions indicated the presence of all members of the family in the cytosolic and purified nuclear fractions. These proteins accumulated to 0.9% of soluble proteins after 21 days of cold acclimation in winter wheat. This represents a cellular concentration of 1.34 microM. Immunohistochemical localization showed that these proteins are highly expressed in the vascular transition zone. No detectable expression was found in mature xylem, in the shoot apical meristem or lateral root primordia. This differential tissue expression suggests that the sensitive cells near the regions where water tends to freeze first require a higher amount of these proteins. This observation is consistent with the fact that regrowth after freezing stress is highly dependent on the viability of this region of the crown. Electron microscopy analysis using immunogold labelling showed that these proteins are present in the cytoplasm and in the nucleoplasm. They are not found in cell walls or other organelles. In vitro cryoprotective assays indicated that the WCS120 protein (PD50 of 10 micrograms ml-1 or 0.2 microM) are as effective as BSA and sucrose (at 250 mM) against freezing denaturation of lactate dehydrogenase. These results suggest that this protein family may be involved in a general mechanism of protection in the soluble fraction of the cell. Their presence in the nucleoplasm may also suggest a possible protective function of the transcriptional machinery. The high hydrophilicity, the abundance and stability of these proteins to boiling suggest that they may provide a particular micro-environment needed for cell survival in the sensitive vascular transition zone during freezing stress. PMID- 7496406 TI - Effective use of robots as mechanized couriers at Stanford University Hospital. PMID- 7496405 TI - Sequence-based identification of T-DNA insertion mutations in Arabidopsis: actin mutants act2-1 and act4-1. AB - A method is presented to facilitate the isolation of mutations in plant genes, which requires knowledge of the target gene or protein sequence, and is independent of mutant phenotype. The polymerase chain reaction was used to amplify the junctions between a T-DNA insert and the gene of interest from pools of mutant plant lines. The approach was used to identify mutations in Arabidopsis thaliana actin genes. The Arabidopsis genome encodes 10 actins in six ancient subclasses each with distinct expression patterns. Primers in the T-DNA border and highly degenerate actin primers, designed from conserved amino acid motifs, were used to prime the amplification. The PCR products were transferred to filters and probed for actin at low stringency. Thus, mutations in all 10 actin genes were screened for simultaneously. Mutations in the vegetative constitutive actin gene, ACT2, and the pollen-specific actin gene, ACT4, were identified in a population of 5300 lines containing approximately 1.5 T-DNA insertions per line. The screen was sensitive enough that actin insertion alleles were easily distinguished among pools of 100 plant lines. PCR techniques were used which accelerated the purification of mutant lines, and segregation, physical mapping, and sequencing of the act2-1 and act4-1 mutations. This strategy should be generally useful in screening mutant libraries made with a variety of plant insertion elements for mutations in any known sequence. PMID- 7496407 TI - Near-infrared spectrophotometry data collection faults due to fiberoptic failure. PMID- 7496408 TI - Group processes of decision making for hospital-based technology assessment committees. AB - There are a variety of group-judgment methods to resolve controversial issues in health care. Meta-analysis and group judgment methods such as consensus conferences are attempts to bring diverse elements of information together for synthesis. Leape notes that a significant body of literature exists regarding the techniques used to elicit opinions from groups. Organizational structures and functions of groups vary in terms of the natures of interactions among group members and the manners in which final conclusions are reached and expressed. The introduction of the process of technology assessment into the hospital setting introduces a problem inherent in the introduction of somewhat academic processes into the operational real world of interpersonal relations, administrative and medical staff interactions, staff costs, and institutional priorities. Hospital administrative processes are based on the committee approach. Medical staff credentialing, drug formularies, and administrative policies are all developed, approved, and implemented through committees. It would seem logical that if technology assessment is to be effective in the hospital setting, then those same group decision processes inherent in committees should be used in technology assessment. Relatedly, if technology assessment is to be successful in the hospital setting, then how can the limited resources of hospital-based staff be best utilized to carry through the assessment of elected technologies? This paper discusses group decision processes, particularly as they relate to technology assessment. The processes of particular interest are those that focus on group interactions rather than theory-based decision processes. The purpose for the paper is to provide to clinical engineering management and senior hospital management background information to use in the formulation of the operating parameters of a hospital-based technology assessment committee. PMID- 7496409 TI - A low-hemolysis blood aspirator conserves blood during surgery. AB - Blood damage caused by traditional vacuum-operated suction tubes, particularly when air is aspirated along with the blood, usually exceeds damage from all other components. In addition to platelet injury, there is a high degree of hemolysis, which leads to high plasma hemoglobin levels and reduces the number of red blood cells available for reinfusion during cases of blood conservation, such as autologous transfusion and cardiac bypass. This work was undertaken to minimize hemolysis, and the accompanying platelet destruction, during aspiration, with the design of a jet-driven aspirator that separates and removes air from blood immediately within the suction tip. The jet-driven aspirator can suction blood at a range of rates from 100 to at least 700 ml/min, separates and removes 80-100% of aspirated air, operates at any orientation, and generates subatmospheric pressures on the order of only 1 inch H2O. In-vitro hemolysis testing showed a significant reduction in average plasma hemoglobin release, from 19.4 mg/dl to 1.8 mg/dl, when air was removed during blood aspiration. In comparative testing with a conventional vacuum suction tube, the jet-driven aspirator showed significantly less hemolysis than the conventional aspirator at comparable rates of air and blood aspiration. PMID- 7496410 TI - A remote method for simultaneous measurement of corneal thickness and curvature at a single point. AB - Advances in refractive surgery have been limited by the measurement technology for determining corneal thickness and curvature. A measurement technique is needed that can provide a detailed corneal thickness and curvature model without contacting the cornea or obstructing the view of the surgeon or surgical equipment. The authors present preliminary results of a method to remotely measure the thickness and curvature of the human cornea at a single point. The method combines ray tracing and interferometry to estimate thickness and curvature in two orthogonal planes in an area less than 100 microns in diameter. This technique has been successfully used to provide very accurate estimates of several thin-shelled test objects. Based upon these results, recommendations are given for further improvement of the technique and extension to a multipoint cornea-modeling system. PMID- 7496411 TI - Antigenic proteins in latex glove extracts: sources of variability. AB - Antigenic protein levels in commercially available latex medical gloves were studied. For comparison, the gloves were divided into four main groups; powdered examination, powder-free examination, powdered surgical, and powder-free surgical. Residual protein levels of 91 different glove brands were determined using the LEAP (latex ELISA for antigenic proteins) assay and found to be extremely variable. The possible sources of variability were investigated. Lot-to lot variability was determined by testing multiple lots of five different brands of surgical gloves. The authors also determined glove-to-glove variability, variability within an individual glove, and test-method variability. It was observed that lot-to-lot variability (25-61% relative standard deviation--RSD) and glove-to-glove variability (51% RSD) were of similar magnitude and accounted for most of the variability found within a given brand of gloves. However, the greatest source of variability identified was brand-to-brand variability (143 283% RSD). PMID- 7496412 TI - The challenges and opportunities of creating an integrated global market for medical devices. PMID- 7496413 TI - Ventilators. PMID- 7496414 TI - A new clinical engineering curriculum. PMID- 7496417 TI - [Is sperm analysis of any value in the management of early, spontaneous abortions?]. PMID- 7496416 TI - [Adenomyosis: role of hysteroscopic treatment]. PMID- 7496415 TI - [Advantages and inconveniences of consensus conferences]. PMID- 7496418 TI - [History of sterility in ancient times. II. Sterility in Hippocrates treatise]. AB - Infertility has always been a constant preoccupation. Hippocrates was the first to write a treatise on infertility. In this historical study we analyse the physiopathology, the diagnosis and the treatment of infertility in the hippocratic's corpus. PMID- 7496419 TI - [Male contraception by hyperthermia]. AB - Assessment of the contraceptive efficacy of a daily mild increase (1 degree C to 2 degree C) in testicular temperature during waking hours is reported in nine couples using two techniques of immobilization of the testes in a supra-scrotal position (close to the inguinal canal). With technique 1 (3 couples and 42 cycles), immobilization was achieved with a specific underwear; there was one undesired pregnancy from a man who stopped the heating for 3 weeks. With technique 2 (6 couples and 117 cycles), immobilization was achieved by adding a supplementary ring to the specific underwear; there was no pregnancy. Reversibility and safety were evaluated. These results suggest that a daily mild increase in testis temperature could be a potential contraceptive method for men. PMID- 7496420 TI - [Early immunosuppression and implantation]. AB - In this brief review, we recall the antigenic status of the embryo and the possible threats that the maternal immune system poses to fetal survival even at implantation stage. We then quickly recall the existence of Early Pregnancy Factor (EPF), the available data for early embryo derived suppressor and suppressor inducer molecules, point to the existence of cloned progesterone dependent T cell derived suppressor molecule, disquisite on immunoregulatory properties of tau interferons, and finish by dealing with Interleukin 10 and the TH1/TH2 balance concept. PMID- 7496421 TI - [In vitro modulation of the production of the cytokine HILDA/LIF, secreted by human endometrial explants: preliminary results]. AB - Human endometrium from fertile women secrete HILDA/LIF in vitro. In endometrial explants from women suffering from an unexplained sterility or women whose fertilized eggs do not implant the in vitro secretion is significantly reduced. HILDA/LIF secretion is increased by IL-1 and decreased by IL-4. CSF-1 has little effects on secretion from fertile women but clearly inhibits secretion from some infertile women. Altogether, the results suggest that HILDA/LIF might be involved in human implantation. PMID- 7496422 TI - [The interleukin-1 system during human implantation]. AB - Although the immune and reproductive systems have been considered independent of each other, the cooperation of both systems are now known to be crucial for the initiation and maintenance of mammalian pregnancy. Nowadays cytokines and growth factors have became increasingly implicated in embryonic implantation. Endometrial functions, embryonic secretions and embryo-endometrial interactions require a continuous dialogue and synchronism between both partners (endometrium and embryo). The present review focuses on the Interleukin-1 system as an example of local regulator in embryonic implantation. Evidence demonstrating its presence and relevance on human endometrium physiology and human preimplantation embryonic development are presented. Furthermore, we described data suggesting the possible role of this cytokine in human implantation. PMID- 7496423 TI - [Tumor necrosis factor and underlying infection]. AB - Tumor necrosis factor alpha (TNF alpha) is a cytokine with pleiotropic actions on several cell types. It is produced particularly by macrophages and by several cells not classically associated with the immune system. Many infections increase the production of TNF alpha; specially in the endometrium where too more TNF alpha can induce failure of implantation. A treatment using antibiotics and antiprostaglandins seem necessary to stop inflammatory process and to increase chance of pregnancy. PMID- 7496424 TI - [Is male fertility threatened?]. PMID- 7496426 TI - [Study of uterine blood supply by pulsed doppler before embryo transfer]. PMID- 7496425 TI - [Operative hysteroscopy in 1995. Equipment, technique, indications and results]. PMID- 7496428 TI - [Rate of precocious sexual intercourse in adolescents in disadvantaged socioeconomic environments. (University of Quebec in Montreal)]. AB - This article presents a preliminary study, a descriptive one, on the sexual intercourse precocity rates at adolescence. 765 male adolescents francophones, 13 years old, from low economic status answered the QAS, questionnaire on aspects of sexual life including. The results show that 11.3% of subjects are sexually active by 13 years old. The first sexual partner is known from many months to many years from the majority of subjects. The majority (61.8%) of the non sexually active subjects of the research would be near to transit to the first sexual intercourse if we consider the results of our predictive measure of the transition. Discussed at the light of the literature, these results show that early sexual behaviors are: 1) a concrete reality for an important proportion of male adolescents from low economic status; 2) to consider in the future in terms of research priority. PMID- 7496427 TI - [Biological status of adolescent girls using estrogen-progestins]. AB - Frequently adolescents use oral contraceptives (OP). Measures of some biological and metabolical patterns were performed in a sample of 339 young girls under going periodic health examination at Pasteur Institute in Lille. A case control study showed lower glucose level, and lower quantity of red blood cells and hemoglobin in OP users. Variance analysis showed lower GGT and TGO with "Gestodene", "Desogestrel" and "Norgestimate" than with the older progestatives (but cigarette smoking seems to influence these datas). Conclusion claims that new progestatives have the best biological tolerance. PMID- 7496429 TI - [Regulation by maternally derived cytokines of pre-implantation development and uterine receptiveness]. AB - Increasing evidence is revealing that cytokines, under the control of steroid hormones, are important in regulating fetal-maternal interactions during pregnancy. Mice carrying mutated genes coding for these factors or their receptors, have been instrumental in clarifying the in vivo roles of these factors in reproduction. Lack of many of the cytokines expressed in the male and female reproductive tracts, such as TNF-alpha, Inf-gamma, the majority of the interleukins and the transforming growth factors TGF-alpha and beta do not result in any gross impairment in reproductive efficiency. However, some cytokines do have a critical role, in particular Leukemia Inhibitory Factor (LIF), Colony stimulating Factor-1 (CSF-1), Epidermal Growth Factor (EGF) and possibly interleukin-1 (IL-1). The recent advances relating to the reproductive roles of these factors up to the time of embryo implantation will be discussed. PMID- 7496430 TI - [Trophoblastic interferons and embryonal immune tolerance]. AB - Current evidence support the hypothesis that trophoblast interferons play a key role in preventing maternal immunologal rejection of the embryonic semi allograft. The information of this review is divided in two sections. In the first section we described molecular and biological characteristics of type I (alpha, beta, omega, tau and spl) and type II (gamma) interferons. In the second section we emphasize studies on immunoendocrine functions of IFN-tau (oTP-1 or trophoblastins) in the network of cytokines and hormonal environment at the uterine embryonic interface. PMID- 7496431 TI - [Adhesion molecules and trophoblastic invasion]. AB - At the time of implantation, the extracellular matrix proteins laminin and fibronectin are abundant in the decidua and are distributed pericellularly around each individual stromal cells. First trimester human trophoblast express both laminin and fibronectin receptors, specifically the alpha 1 beta 1, alpha 5 beta 1, alpha 6 beta 1 and alpha 6 beta 4 integrin heterodimers. We have demonstrated that in vitro adhesion of first trimester human trophoblast to purified extracellular matrix proteins and to purified decidual stromal cell monolayers can be inhibited by monoclonal antibodies directed against appropriate integrin subunits and by synthetic peptides containing an RGD sequence. We have also examined the downstream events of trophoblast matrix adhesion, such as integrin mediated signal transduction and the production of metalloproteinases. The implications of these interactions are discussed. PMID- 7496433 TI - The prevention of human transmission of gastrointestinal infections, infestations, and bacterial intoxications. A guide for public health physicians and environmental health officers in England and Wales. A working party of the PHLS Salmonella Committee. PMID- 7496432 TI - [Expression and role of complement regulatory proteins on human gametes and pre implantation embryos]. AB - Human gametes and pre-implantation embryos express selectively several complement regulatory proteins. Membrane cofactor protein (MCP, CD46) and decay accelerating factor (DAF, CD55) are regulators for C3 convertases and protectin (CD59) is an inhibitor of the membrane attack complex. These three proteins were identified on human sperm and found to be functional. CD55 and CD59 were both expressed by the plasmic membrane of unfertilized oocytes and pre-implantation embryos. CD46 was not present on unfertilized oocytes but appeared at the 6/8 cell-stage embryo when human gene expression first occurs. Complement receptor 1 (CR1, CD35) and MHC class I antigens were not found on oocytes neither on embryos. Such a selective expression of complement regulatory proteins associated with the lack of MHC class I antigens may represent an immune protective mechanism by which human gametes and pre-implantation embryos escape from complement-mediated damage during their travel through the female genital tract. Indeed uterine, tubal and follicular fluids contain all the components of the complement cascade, including classical and alternative pathways. Nevertheless participation of CD46 and CD59 in cell to cell interaction during fertilization and/or implantation cannot be excluded. CD59 is an adhesive molecule involved in the rosette phenomena and CD46 has been described as the human receptor for measles virus, which binds through a fusion protein. Monoclonal antibodies raised against these two proteins (CD46 and CD59) are able to inhibit heterospecific fertilization between zona-free hamster oocytes and human spermatozoa suggesting the role of these proteins during fertilization. PMID- 7496434 TI - Dengue: current epidemics and risks to travellers. PMID- 7496435 TI - Combined drugs may be better than zidovudine alone for immunocompromised adults infected with HIV-1. PMID- 7496436 TI - AIDS and HIV-1 infection in the United Kingdom: monthly report. PMID- 7496437 TI - Influenza surveillance: 1995/96. PMID- 7496438 TI - A prospective long-term follow-up study of patients evaluated for erectile dysfunction: outcome and associated factors. AB - The aim of this study was to investigate the long-term outcome of erectile dysfunction (ED) assessed in a routine clinical setting. The original population consisted of 209 patients who were consecutively referred to either the Urology Clinic or the Sexual Dysfunction Clinic of a University Hospital. At follow-up, 22 patients had died and 32 could not be found. Of the remaining 155, 107 (69%) patients participated in the study. The mean follow-up period was 4.1 years. Of these, 21 had had a prosthesis implanted, 34 had received self-injection therapy. 31 had sex therapy and 28 patients had not received any treatment. Overall, the rate of penetration increased, whereas coital frequency did not change. Despite the fact that sexual functioning in terms of penetration rate improved, more than of the patients reported that they were dissatisfied with the overall quality of their sex-lives. PMID- 7496439 TI - Effect of vasoactive intestinal peptide and acetylcholine on penile erection in the rat in vivo. AB - We investigated the effect of vasoactive intestinal peptide (VIP) and its combination with acetylcholine (ACh) on the erectile response in 52 adults rats. Intracavernous injection of VIP (10(-8) to 10(-5)M, ACh (10(-9) to 10(-5), or a combination of VIP (10(-7) to 10(-5) with ACh (10(-6 M) additively enhanced that erection but did not lead to a full erection. VIP-antagonist (10(-9) to 10(-5 M), as well as atropine alone (10 (-7) to 10(-5 M), partially suppressed full erection induced by cavernous nerve stimulation (1 Hz, 3-6 V) in a dose-dependent manner, and the combination of VIP-antagonist (10(-9) to 10(-5 M) with atropine (10(-6 M) showed an additive effect. The results indicate that although VIP as well as ACh in involved in penile erection, they are not likely to be principal neurotransmitters. Their clinical application in the treatment of impotence may be confined to use in conjunction with another vasoactive agent. PMID- 7496440 TI - Corporeal plication for surgical correction in Peyronie's disease improves rigidity. AB - A prospective study was done on 76 patients who underwent surgical correction by corporeal plication of the penile deformity secondary to Peyronie's disease. Nocturnal penile tumescence and rigidity were recorded by RigiScan during three consecutive nights, before and after the operation. A significant number of patients developed erections of better quality especially at the tip, after this significantly enhance the quality of erectile episodes, by increasing penile rigidity, during rapid eye movement sleep. PMID- 7496441 TI - The clinical assessment of erectile dysfunction: a comparison of nocturnal penile tumescence monitoring and intracavernosal injections. AB - One hundred and fifty-nine men with erectile dysfunction were assessed with Rigiscan monitoring of nocturnal penile tumescence (NPT) and response to intracavernosal injections (ICI) of papaverine or prostaglandin E1. A satisfactory NPT, suggestive of psychogenic causation, was recorded in 58%, whereas 15% had clearly impaired NPT. There was a significant association between presence of vascular disease, diabetes and impairment of NPT. For ICI, only 32% showed a satisfactory response, with 48% clearly impaired. Of the 92 men with satisfactory NPT, 40% HAD clearly impaired and 41% satisfactory ICI response. Of the 51 men with satisfactory ICI response, 8% had clearly impaired and 74.5% satisfactory NPT. There was no association between ICI response and history of vascular disease. We conclude that monitoring of NPT by Rigiscan, and without sleep monitoring, is a valuable diagnostic procedure. In contrast, because of the high proportion of false negative results. ICI monitoring is of very limited diagnostic value. The explanation for false negative ICI responses, however, could prove to be of considerable theoretical and clinical importance. PMID- 7496442 TI - Effects of priapism lasting 24 hours or longer caused by intracavernosal injection of vasoactive drugs. AB - Twenty-four patients, treated after > or = 24 h of priapism caused by the injection of vasoactive drugs, were examined. The aetiology of impotence was psychogenic in > 50% of the patients (13/24). Most priapisms that lasted < 36 h could be treated successfully by puncture and alpha-adrenergic drugs without any fibrosis of the corpora cavernosa, and most patients continued to receive vasoactive agents for the treatment of impotence; but this approach was no longer adequate after priapism had lasted > = or 36 h when glandulocavernosal shunts were needed to achieve permanent detumescence. After > 48 h of priapism all the patients developed fibrosis of the corpora cavernosa and none of them was able to continue with the injections of vasoactive drugs. In addition to fibrosis of the penis, one fulminant infection, one case of urethral damage, one permanent shunt and two cases of transient hypertonia with ECG changes were seen. In order to save time, glandulocavernosal shunts should be performed without delay once priapism has lasted > 36 h. PMID- 7496443 TI - Elastic fibre concentration in the tunica albuginea of corpora cavernosa and nocturnal tumescence monitoring. AB - The tunica albuginea of the corpus cavernosum provides the latter with a fibrous framework and plays a significant role in erectile function. Being rich in elastic fibres the tunica albuginea is able to resist overstretching of the corpus at raised levels of intracavernous pressure, compressing the sub algunineum venous reticulum and promoting the maintenance of erection. Results are reported here on assessment of the concentration of elastic fibres in tunica albuginea in relation to frequency of nocturnal erection, tumescence and penile length and rigidity. Significant correlations were demonstrated between concentration of elastic fibres and duration of nocturnal erection (P< 0.0001), rigidity at TIP(P< 0.001), and rigidity at BASE (P< 0.001). The importance of the structural soundness of the tunica albuginea for achievement of satisfactory erection was thereby underlined. PMID- 7496444 TI - Analysis of continuous nocturnal penile rigidity measurements with the use of the RigiScan summary analysis software program. AB - The evaluation of the RigiScan nocturnal recordings of 324 patients, 18 to 72 years old (mean age 49 years), according to guidelines based on normal data, were compared with analysis of these recordings by using the RigiScan Summary Analysis Software Program. This enabled us to provide normal values for this program. We conclude that it should be possible to extend this RigiScan software program in such a way that a probability of normality is given to its user. In patients with corporal veno-occlusive dysfunction a significant lower total event duration was found when compared with the results of patients with normal and abnormal findings. PMID- 7496445 TI - An attempt to standardize the pharmacological diagnostic screening of vasculogenic impotence with prostaglandin E1. AB - The response to the intracavernosal injection of differing doses of prostaglandin E1 (PGE1)(10, 20 and 30 micrograms) was compared in a randomized prospective manner in 283 men with chronic impotence, in an attempt to identify the optimal drug dose for pharmacological diagnosis. The mean age of patients studied was 63.1 years, and comprised 90 men with arteriogenic impotence, 133 men with either pure venogenic impotence or venous leakage associated with arteriogenic impotence, and 60 men with psychogenic impotence. Erectile responses were quantified using real-time RigiScan monitoring and were compared to a diagnosis based on the patient history, examination findings and all subsequent investigations. In patients with arteriogenic impotence, correct pharmacological diagnosis with doses of 10, 20 and 30 micrograms of PGE1, was made in 71, 89 and 90% of patients respectively. In patients with venous leakage, correct pharmacological diagnosis with doses of 10, 20 and 30 micrograms of PGE1 was made in 95, 95 and 93% of patients respectively. In patients with psychogenic impotence, correct pharmacological diagnosis with doses of 10, 20 and 30 micrograms of PGE1 was made in 72, 95 and 98% of patients respectively. This study indicates that pharmacological diagnosis with PGE1, is a useful screening test to differentiate penile vascular disease from psychogenic impotence and arteriogenic impotence from cavernosal venous leakage, achieves optimal sensitivity and specificity with doses of 20 micrograms or greater, but is associated with a high false diagnosis rate of diagnosis of penile vascular disease if doses of less than 20 micrograms are used. PMID- 7496446 TI - Effects of papaverine and vasointestinal polypeptide on penile and vascular cAMP and cGMP in control and diabetic animals: an in vitro study. AB - Adenosine 3'5'-cyclic monophosphate (cAMP) and guanosine 3'5'-cyclic monophosphate (cGMP) mediate penile erection. We have previously established that adenylate and guanylate cyclase activity is elevated in the diabetic rat penis and aorta. This study investigates the action of papaverine and vasoactive intestinal polypeptide (VIP) on these cyclases. The aortae and penes of Sprague Dawley rats (n = 7) were stimulated with VIP and papaverine. Diabetes mellitus (DM) was induced in Sprague Dawley rats (n = 7) with streptozotocin and the penile and aortic tissues were treated with VIP. The penes, aortae and carotid arteries of New Zealand White rabbits were similarly processed. cAMP and cGMP generation was measured by radioimmunoassay. In all tissues: VIP stimulated cAMP synthesis; VIP did not increase cGMP levels; papaverine was without effect on either cAMP or cGMP synthesis. VIP-stimulated cAMP was significantly enhanced in the diabetic rat penis and aorta; there was also a significant elevation in the basal levels of cGMP in these tissues. These data: (1) consolidate that cAMP is a mediator of penile erection, (2) indicate that papaverine and VIP elicit erection by different mechanisms, (3) suggest that an enhanced penile capacity to generate cAMP in DM may constitute an adaptive response to counteract the previously reported reduction in VIP content and VIP receptors, and (4) indicate that the penile and vascular tissues of the rabbit respond in a similar manner to VIP and papaverine. PMID- 7496447 TI - Distinct behavior of portal venous and arterial vascular waterfalls in porcine liver. AB - PURPOSE: Hepatic dysfunction is associated with morbidity and mortality in critically ill patients. Understanding liver hemodynamics in pathological states requires characterization of the normal portal venous and hepatic arterial circulations. Using pressure flow analysis, we tested the hypothesis that vascular waterfalls determine blood flows in the normal liver. METHODS: In 14 vascularly isolated porcine livers, steady-state pressure-flow relationships, which defined a slope (incremental resistance) and a zero flow pressure intercept (Po), were generated for each vessel over a range of hepatic venous pressures (Phv). RESULTS: Critical closing pressures occurred in the portal venous circulation (Po = 3.8 +/- 0.4 mm Hg) with classical waterfall physiology observed as Phv was raised. The hepatic arterial critical closing pressure (Po = 8.3 +/- 1 mm Hg) showed a constant positive pressure difference of mm Hg versus Phv as the latter was increased from 0 to 28 mm Hg (P < .05). Portal venous resistance decreased when Phv was greater than Po (P < .05), but no effect on hepatic arterial resistance was seen as Phv was increased. CONCLUSION: Both critical closing pressures and incremental resistances showed markedly different responses to increased outflow pressures in the portal venous and hepatic arterial circulations. The results provide the physiological basis to analyze hemodynamic changes in the liver under normal and pathological conditions. PMID- 7496448 TI - Inspiratory tidal volume sparing effects of tracheal gas insufflation in dogs with oleic acid-induced lung injury. AB - PURPOSE: Tracheal gas insufflation (TGI) improves the efficiency of conventional mechanical ventilation (CMV) by reducing the series dead space of the airways. Consequently, application of TGI as an adjunct to CMV may permit reducing tidal volume (VT) while limiting CO2 retention. We tested the extent to which panexpiratory TGI allows reduction of VT while maintaining PaCO2 constant in an oleic acid-induced lung injury model. METHODS: We studied six anesthetized, paralyzed, and mechanically ventilated dogs. Oleic acid injury was induced by injecting 0.09 mL/kg of oleic acid into the right atrium. After stabilization of lung injury the VT-sparing effect of TGI was tested by progressively increasing catheter flow rate (Vc) from 2 to 5, 10, and 15 L/min while decreasing VT by an amount that maintained PaCO2 constant (approximately 47 mm Hg) with respect to baseline (Vc = 0 L/min). RESULTS: Tidal volume was decreased from a baseline value of 0.360 +/- 0.030 L to 0.238 +/- 0.054 L at Vc of 15 L/min. The reduction in VT was associated with a decrement in peak and end-inspiratory plateau airway opening pressure from 32 +/- 3 to 28 +/- 6 cm H2O and from 25 +/- 2 to 21 +/- 3 cm H2O, respectively. Total physiological dead space fraction decreased from a baseline value of 0.60 +/- 0.08 to 0.31 +/- 0.20 during TGI at 15 L/min. TGI did not affect cardiac output, PaO2, or pulmonary venous admixture. CONCLUSION: We conclude that TGI can be a useful adjunct to CMV during acute lung injury to limit VT while avoiding CO2 retention. PMID- 7496449 TI - Oxidative metabolism in sepsis and sepsis syndrome. AB - The high mortality associated with sepsis syndrome and multiple organ dysfunction syndrome has persisted despite extraordinary research efforts in the laboratory and the intensive care unit. These syndromes produce systemic tissue damage that is likely to result from widespread inflammation and subsequent endothelial injury. This article reviews the oxidative metabolic effects and responses to sepsis syndrome at several levels: the oxygen transport system, the cell, and the mitochondrion. Specifically, aerobic metabolism of carbon substrates and oxygen is altered in sepsis. As a result of systemic inflammation and nonmetabolic oxygen use, oxidative stress may occur both outside and inside the cell. The consequences of these oxidative processes during sepsis may be ongoing cell damage mediated by reactive oxygen and nitrogen oxide species that culminates in multisystem organ failure. PMID- 7496450 TI - Current techniques in cell and molecular biology. AB - Recent advances in the field of molecular biology have revolutionized our understanding of the functioning of living organisms and facilitated the development of robust tools for both diagnosis and treatment of diseases. With particular reference to the field of critical care medicine, development of molecular biology techniques have aided in the following: (1) rapid and highly specific detection of pathogenic infectious agents (eg, Mycobacterium tuberculosis, Pneumocystis carinii, cytomegalovirus, Legionella); (2) development of assays for measurement of circulating cytokines such as tumor necrosis factor (TNF) and interleukin (IL)-1 that has helped our understanding of the pathogenesis of the sepsis syndrome; (3) administration of antibodies or soluble receptors to attempt to prevent untoward effects of cytokines such as TNF or IL 1; and (4) the administration of recombinant deoxyribonucleic acid (DNA) or proteins to patients in an attempt to alter the course of a disease such as antioxidant enzymes (superoxide dismutase). The rapidity of progress in this field has been staggering, which necessitates frequent updating of our knowledge for clinicians to put these molecular tools to their best use. This brief review attempts to explain the basic principles of commonly used techniques in molecular biology including recombinant DNA, polymerase chain reaction, DNA libraries, gene therapy, and protein biochemistry in a manner that is understandable to those without an in-depth knowledge of the field. PMID- 7496452 TI - Innovations in patient care: changing clinical practice and improving quality. AB - BACKGROUND: In the traditional hospital organization, administrators supply the resources while physicians determine their use. Given this dichotomy, a partnership between clinicians and hospital management is essential for efforts to enhance the quality of care while controlling costs. To foster this partnership, in 1986 the University of Rochester's Strong Memorial Hospital developed its Innovations in Patient Care (IPC) program, which several other medical centers have duplicated. CURRENT STATUS AND LOGISTICS: Hospital operating revenues of approximately $175,000 per year are provided to both fund proposals and support core IPC staff. Clinical staff submit proposals to study innovations to promote higher quality care and/or the efficient and appropriate use of diagnostic and therapeutic services. Many of the 77 projects funded to date have led to important changes in clinical practice. CASE STUDIES: One study, whose principal investigator was assistant director of emergency medicine, showed that structured, condition-specific (for example, asthma, pharyngitis, lacerations, and isolated closed-head injury) quicksheets improved documentation of clinical findings, resource use, and clinical practice. A study organized by the leadership of surgical nursing revealed that a nursing case management model led to reductions in patient length of stay and increases in nurse satisfaction. Another study, designed by a fellow in neonatalogy, developed and tested guidelines for the use of head ultrasounds in screening very-low-birthweight infants for intraventricular hemorrhage. CONCLUSIONS: IPC programs, which integrate well with initiatives in total quality management, can be effectively used to change clinical practice and improve the quality and efficiency of patient care. PMID- 7496451 TI - Continuous oscillation: outcome in critically ill patients. AB - PURPOSE: To compare turning by an oscillating bed to standard 2-hour turning. Outcomes were survival, length of stay (LOS), duration of mechanical ventilation, and incidence of pneumonia. METHODS: One hundred and three intensive care patients were randomly assigned to standard turning or turning by an oscillating bed. Data, collected at baseline, daily for 7 days, and then three times weekly until study discharge, included demographics, initial Acute Physiology and Chronic Health Evaluation (APACHE II) score, ventilatory/gas exchange parameters, indicators of pneumonia, nursing measures, and chest roentgenograph. RESULTS: There were no significant differences for LOS, duration of ventilation, nor incidence of pneumonia. Higher survival for subjects on the oscillating bed reached borderline significance (P = .056) for subjects with APACHE II greater than or equal to 20. Longitudinal data were analyzed using the random effects model. No differences in ventilatory or gas exchange parameters were identified. Among subjects who developed pneumonia there was a significantly higher respiratory score (nursing acuity scale) for subjects on the oscillating bed. CONCLUSIONS: In selected critically ill patients oscillating therapy may improve survival and improve airway clearance. The frequency and degree of turning needed to prevent complications and improve outcome remains unclear. These newer beds should be used with discrimination so as to not increase hospital costs unnecessarily. PMID- 7496453 TI - The Benchmarking Effort for Networking Children's Hospitals (BENCHmark). AB - BACKGROUND: In 1992, 12 large children's hospitals established the Benchmarking Effort for Networking Children's Hospitals (BENCHmark). The goal was for the BENCHmark effort to supplement the hospitals' continuous quality improvement (CQI) programs and to speed adoption of best practices from peer institutions. For three years, the hospitals have been comparing data on cost, quality, and speed indicators. Also, "best practice" groups have met to share information on how processes can be improved. RESULTS: The BENCHmark hospitals have experienced significant process improvement in areas such as emergency department waiting time and admitting process time. EXAMPLE: The BENCHmark hospitals selected admitting as one of the first best practice groups to meet. Interdisciplinary staff from all BENCHmark hospitals met three times over the course of a year to define their indicator and share information on best practices. St Louis Children's Hospital, as a result, instituted a pre-arrival team and cross-trained staff, with the result being a reduction of admitting processing time from 58 minutes to 19 minutes. Same-day surgery patients now bypass the admitting department and go directly to the surgical floor. Patient and surgeon satisfaction has increased greatly. CONCLUSIONS: Hospitals that are planning to benchmark are encouraged to reach consensus on project goals and to focus on indicators that provide a clear business advantage. Physician involvement is key to improving performance and physicians will only be engaged if the hospitals against whom they are benchmarked are considered peers. Being willing to share initial data openly seems to be a key factor in determining successful integration of the BENCHmark process into hospital CQI efforts. The BENCHmark project has been so successful that a second group of 12 comparable pediatric institutions, known as the Network II, has been established. PMID- 7496455 TI - Increasing the impact of quality improvement on health: an expert panel method for setting institutional priorities. AB - BACKGROUND: Successful implementation of modern ongoing quality improvement (QI) methods requires investment of institutional resources, but can produce significant improvements in medical care. A health care organization's goals and objectives for improving care are expressed in strategic plan documents, which could provide a framework for planning quality improvement initiatives. However, institutional strategic planning processes are often not well linked to QI staff and resources. We developed the Quality Action Program (QAP) to connect QI to strategic planning. HISTORY: In 1991, Sepulveda VHAMC implemented a major primary care initiative, documented in a comprehensive strategic plan. The QAP was developed to enable the initiative to be evaluated within a QI context. THREE ROUND EXPERT PANEL PROCESS: To carry out the QAP, members of an institution's quality council engage in a structured consensus process. The first round involves reading educational materials and filling out a quality action survey the second round includes participation in an expert panel meeting, and the third round involves making final priority rankings. EIGHT-STEP QAP IMPLEMENTATION PLAN: QI staff carry out activities to prepare for and carry out the three-round expert panel process. RESULTS: QAP induced significant institutional QI activity directed toward achieving the top-ranked QI criterion--ensuring continuity of care. Continuity of care improved significantly over time between the pre- and post-QAP periods. CONCLUSIONS: Expert panel methods can be used to link strategic plan goals and objectives to QI efforts. PMID- 7496454 TI - Variability in clinical systems: applying modern quality control methods to health care. AB - BACKGROUND: Quality health care depends on timely completion of sequences of clinical care. This study evaluated the concepts and tools of quality management for measuring system performance in ambulatory care. STUDY DESIGN: Clinical staff in nine centers of a group model health maintenance organization described the procedure for ordering and completing a complete blood count, mammogram, and surgical consultation. Variability was noted among the processes as intended and as actually performed, as well as inconsistencies reported within and among centers. In two centers investigators tracked performance of key sequences of care and the achievement of other key quality characteristics believed desired by physicians and patients. Computerized patient records and departmental files were the only available sources of data for assessing completion and followup of tests and consultations. Even these data were difficult to obtain and, in many instances, incomplete. RESULTS: Although data were often difficult to obtain, the quality management techniques used were helpful in revealing process failures that appeared to be the result of design flaws built into the clinical systems. CONCLUSION: Robust process designs and improved management information systems for monitoring these processes are recommended to reduce variability and improve the quality of clinical care. PMID- 7496456 TI - [Treatment of burns in infants]. AB - Because of the potential severity of their residual deformities, burn injuries in infants justify an early management in specialized centres when they cover more than 5% of body surface and in every case when hands, face, or external genitalia are concerned. Cooling with cold water is the first aid treatment to be performed as early as possible after the injury. The treatment in specialized centres must be both general and surgical. General treatment includes fluid and electrolyte therapy, temperature control, appropriate nutrition and pain suppression. Pain suppression is a major part of the treatment and morphine must be largely used. Surgical treatment starts as soon as the patient arrives in the centre and is eventually performed under general anesthesia: all the burned areas are covered with occlusive dressings. Infections are prevented by systematic cultures and adjusted antibiotic therapy. A vigorous rehabilitation program must be instituted as soon as possible: massages, compressive clothes, splints, physical therapy, plastic surgery. Primary prevention by sustained parental education is important in order to reduce the frequency of burn injuries in infants. PMID- 7496458 TI - [Radiological case of the month. A case with osteocartilaginous exostosis of the distal end of the femur]. PMID- 7496457 TI - [Complementary neuromotor examination of infants at risk for sequelae.Why? How?]. AB - In many cases the traditional neurological examination of high risk newborns for neurological problems secondary to perinatal difficulties does not identify easily the absence or the presence of cerebral palsy before walking. To help the neuromoteur follow up in these children, the authors suggest a complementary neuromotor examination that is successful only in normal infants. This examination is done in front of a mirror in the presence of the parents in order to increase the attention of the child, whose participation is necessary for a successful examination. Two motor responses are evaluated: the lateral support on, alternately, right and left superior limbs, and the lateral abduction reaction of the hip of each limb. When normal these two motor responses allow an early affirmation of integrity of motor paths if the examination is performed under carefully controlled conditions. PMID- 7496459 TI - [Diagnosing neonatal pustulosis: pustulosis induced by Malassezia furfur]. PMID- 7496460 TI - [Systematic screening for neuroblastoma]. PMID- 7496461 TI - [Cocaine and pregnancy: a disturbing association with the increase of addiction]. PMID- 7496462 TI - [Hyperleukocytosis during pediatric emergencies]. PMID- 7496463 TI - [Nosocomial respiratory infection due to coronavirus in neonatal intensive care units: prospective evaluation]. PMID- 7496465 TI - [Bone pain revealing parathyroid adenoma]. PMID- 7496464 TI - [Risk of methemoglobinemia after Emla application in premature infants]. PMID- 7496466 TI - [Value of echocardiography in Ellis-Van Creveld syndrome]. PMID- 7496467 TI - [Syringes for oral administration of vitamin K1 lipid emulsion]. PMID- 7496468 TI - [Prevalence of Schistosoma intercalatum intestinal bilharziasis in children hospitalized the urban area of Gabon]. PMID- 7496469 TI - [The child and his physician]. PMID- 7496470 TI - [Plea for early diagnosis of biliary tract atresia. Twelve errors to avoid]. PMID- 7496471 TI - [Neurological follow-up of newborn infants at risk for cerebral palsy]. AB - BACKGROUND: Traditional neurological examination of newborns at risk for cerebral palsy (CP) is based on periodic assessment of normal milestones so that the diagnosis of CP can be very late. A complementary neuromotor evaluation based on two motor series has been proposed in order to make possible an early affirmation of integrity of motor paths. POPULATION AND METHODS: Eight hundred and seventy three infants including 371 preterms had been admitted to a neonatal intensive care unit from 1990 to 1992. Then, they were examined as outpatients by a multidisciplinary team. They were especially tested for lateral support on right and left superior limbs, alternately, and for lateral abduction reaction of each hip. RESULTS: Absence of manifestations of CP was noted in 87% of infants with a corrected age of 4 months; a correct prediction of future normal walking was made for all babies. Both tests could not be correctly performed by those babies who lately developed manifestations of CP. CONCLUSIONS: These two simple tests appear to be accurate to early predict normality. PMID- 7496472 TI - [French epidemiological survey of burns in children under 5 years of age]. AB - AIMS: To add to epidemiological data concerning serious accidents in the home in France. PATIENTS AND METHODS: This prospective study, carried out over 12 months, included all children with burns admitted to 14 burn units (BU) and 18 pediatric surgery units (PS) using a questionnaire from the SFETB comprising 20 items. RESULTS: On population 0 to 60 months, 687 patients were included (434 in BU, 253 in PS). Descriptive analysis and correlation of several parameters identified patterns of burns in childhood and the factors of severity. Victim data: young boys (59.3%), aged < 36 months (mean = 24 months), burned at home, in the kitchen (62.4%) with hot fluids (73%), or bathroom (16.2%) with tap water by immersion. Flames were cause of burns (8.7%) by inflammable products. Lesions were always more severe in surface and risk of mortality was higher. Burns covered a surface equal or less than 10% in 57% of cases. CONCLUSIONS: The value of the study was to be more aware of the dangers of which children are exposed and to encourage application of preventive measures. PMID- 7496473 TI - [Medical costs associated with the risk of maternal-fetal contamination by HIV. Results of a prospective survey]. AB - BACKGROUND: Assessment of economic implications of pediatric HIV infection allows to document efforts which are devoted by health care system to the care of children born to HIV infected women. This paper presents the results of the first analysis, in the French health care context, of direct medical costs related to HIV mother-to-child risk of transmission. MATERIAL AND METHODS: Statistical prospective analysis was performed on 853 files concerning 152 children followed in three medical centers. Because the medical consumptions data were linked to clinical status, average direct medical costs per medical contact could be defined for each center. RESULTS: Results were consistent with the hypothesis that more medical care is necessary when the clinical picture is uncertain (indeterminate status) or reflects health state deterioration. However, results are very different according to HIV clinical status and disparities may be so large that costs per clinical status hierarchy may be disrupted. Explicative factors for these variabilities are expressed in terms of patient recruitment, medical uncertainty management and resources disponibility. PMID- 7496474 TI - [Heart and sudden infant death. Anatomopathological study of 100 cases]. AB - BACKGROUND: Post-mortem examination of infants with sudden infant death syndrome (SIDS) is essential for understanding the cause and/or mechanism of death. This study aims to evaluate the contributions of heart in SIDS. POPULATION AND METHODS: Between 1981 and 1990, the CHU of Nantes carried out autopsies on 162 cases of sudden infant death syndrome. One hundred files were accepted for this retrospective study because a second recent macroscopic and microscopic (11 samples for each heart) examination of heart was possible. RESULTS: The weight of hearts was not increased, except in one case. Macroscopic examination did not reveal any cardiac abnormalities. Histological examination confirmed the lesions initially observed in 11 cases and probably responsible for their death. CONCLUSIONS: This study underlines the importance of studying numerous samples of each heart of microscopic examination of many sections. PMID- 7496475 TI - [Acute gastric dilatation with superior mesenteric artery syndrome in a young girl with anorexia nervosa]. AB - BACKGROUND: Acute gastric dilatation is a rare complication of anorexia nervosa which may be fatal in case of gastric perforation. Superior mesenteric artery syndrome may be associated with gastric dilatation. CASE REPORT: A 14 year-old girl was admitted suffering from severe anorexia nervosa. Shortly after admission, she complained of abdominal pain related to a bulimic episode a few days before, followed by nausea and vomiting. Distended abdomen and tachycardia suggested acute small-bowel obstruction. A diagnosis of acute gastric dilatation with superior mesenteric artery syndrome was suspected on X-ray examination and confirmed by esophagogastrography after gastric evacuation. The patient improved rapidly under total parenteral nutrition. Upper gastrointestinal study repeated 10 days later showed normal stomach. CONCLUSIONS: Small-bowel obstruction in anorexia nervosa first evokes acute gastric dilatation, and a perforation has to be ruled out by esophagogastrography. Association with superior mesenteric artery syndrome is possible; it does not necessarily lead to surgery. PMID- 7496476 TI - [Severe hyponatremia associated with anorexia nervosa: role of inappropriate antidiuretic hormone secretion?]. AB - BACKGROUND: Anorexia nervosa may be revealed by hyponatremia or associated with it. This hyponatremia can be due to diuretics or laxatives abuse, potomania or inappropriate secretion of antidiuretic hormone as in the case reported here. CASE REPORT: A 15 year-old girl was admitted for anorexia nervosa. She had secondary amenorrhea and asthenia and had lost 14 kg during the preceding 4 months. Her natremia ranged from 112 to 130 mEq/l with normal urinary sodium excretion, serum hypoosmolality (252 mosm/kg) and inappropriately elevated urinary osmolality (698 mosm/kg). Renal, adrenal and thyroid functions were normal. After water loading, patient remained euvolemic, with a negative free water clearance and an increased plasma ADH level with concomitant hyposmolality. A diagnosis of inappropriate secretion of antidiuretic hormone was made, after excluding other conditions such as potomania, drug abuse, vomiting, tumor and chronic lung disease. CONCLUSIONS: Hyponatremia sometimes seen in anorexia nervosa must lead to the search of inappropriate antidiuretic hormone secretion. PMID- 7496477 TI - [Obstruction: a rare complication of congenital giant blue nevus]. AB - BACKGROUND: The blue nevus is an acquired or congenital dermal melanosis, usually small in size. It is sometimes giant and responsible for complications. CASE REPORT: A newborn had a large infiltrated blue news located on buttocks, lombosacral and perianal areas. He was admitted at the age of 3 weeks for an intestinal obstruction which was due to an impassable anal stenosis secondary to the infiltrated nevus. The tumor was excised twice, completed by plastic surgery at the age of 10 months. CONCLUSIONS: This rare congenital giant blue nevus was responsible for an unreported intestinal obstruction. The risk of malignant transformation was another reason for complete excision. PMID- 7496478 TI - [Lymphomatoid papulosis in a child]. AB - BACKGROUND: Lymphomatoid papulosis (LyP), uncommon in children, has a benign clinical course in contrast with a malignant histology. CASE REPORT: A 9-year old boy developed nodular skin lesions for 6 months on the scalp and penis and a papule skin on his trunk. Surgical excision of scalp nodules was performed to prevent necrosis and ulceration of the ear. The biopsy showed a dense mixed cellular infiltrate including histiocytes, lymphocytes and numerous atypical large cells of which a minority expressed the CD30 antigen. PCR showed an oligoclonal rearrangement of the TCR gamma chain gene. Physical examination, bone marrow aspirate and thoracoabdominal CT scans were normal. Three years after surgery, no other lesions occurred. DISCUSSION: Differential diagnosis of LyP is difficult with non Hodgkin's lymphoma, especially CD30+ lymphoma. Prognosis remains the major problem in LyP. Ten to 20% of cases are associated with lymphomas in adults. PMID- 7496480 TI - 14th International Symposium on the Separation and Analysis of Proteins, Peptides and Polynucleotides. Heidelberg, Germany, 2-4 November 1994. PMID- 7496479 TI - [Pediatric aspects of anthracycline cardiotoxicity and practical implications for prevention]. AB - Discovered during the sixties, anthracycline antibiotics are today widely used anti-cancer drugs. Their potentially fatal cardiac toxicity, which is related in part to the total cumulative dose, has been described since 1967. The aim of this paper is to describe their biological and clinical toxic effects on the heart, especially of children, and to propose prevention guidelines. The mechanisms of cardiac toxicity, with their destructive consequences on functional myocytes reserve, are shortly recalled. Acute, sub-acute and chronic clinical aspects of anthracycline's cardiomyopathy are the subject of a literature review. In Pediatric Oncology, the prolonged survival usually observed allows delayed congestive heart failure to occur by myocardial reserve insufficiency, as hemodynamic needs are quickly increasing, especially at the end of the somatic growth. Furthermore, the frequency of cardiac abnormalities is increasing with time after therapy, reaching about half of the explored patients after 15 years. The main known methods to prevent such a toxicity are reviewed. The parcimonious use of anthracyclines is already seen in children. Every method to decrease the maximal plasma concentration of the drug (weekly schedule or prolonged infusion) has to be considered. The active cardioprotectant agent such as ICRF-187, is in clinical development. Detection, prevention, and therapy of cardiac abnormalities, which are likely to precede delayed heart failure, still remains a difficult problem in these more and more numerous children to be cured of cancer. PMID- 7496481 TI - Preparation and characterization of isozymes and isoforms of horse liver alcohol dehydrogenase. AB - The procedure described allows the simultaneous large-scale preparation of the three main isozymes (EE, ES, SS) of alcohol dehydrogenase from horse liver (HLADH) and their subfractions using heat denaturation, ammonium sulfate precipitation, DEAE and CM ion-exchange chromatography as well as AMP-Sepharose affinity chromatography. Typical yields that can be obtained within three weeks are 1.5-2.5 g of EE-HLADH, 300-800 mg of ES-HLADH, 20-400 mg of SS-HLADH and 50 100 mg of EE-HLADH isoforms from 5 kg of horse liver. The EE-HLADH isoform prepared has a pI of 7.8, which is 0.3 pH units lower as compared to the main fraction; the zinc content and number of free sulfhydryl groups are unchanged but matrix-assisted laser desorption ionization mass spectrometry resulted in a molecular mass difference of + 130 to 165 relative molecular mass. From a sugar determination and comparison of its pI with an artificial glycosylation product of the EE-HLADH isozyme we concluded that the isoforms of HLADH are non-enzymatic glycosylation products which have been described to occur during protein aging. PMID- 7496482 TI - New approaches to chromatographic purification of bovine dopamine-beta hydroxylase. AB - The use of the traditional scheme for the isolation of bovine dopamine-beta hydroxylase (bDBH) from bovine adrenal medulla resulted in active but not pure bDBH, containing about 50% of admixtures. Immobilized metal chelate affinity chromatography on agarose modified with iminodiacetic acid residues and charged with cobalt ions was applied in the final stage to obtain more than 90% pure and active bDBH. Final purification of bDBH using step elution with 0-0.5 M methyl-D mannoside in buffer solution from concanavalin A-Sepharose was studied. The determination of bDBH in various samples was performed using size-exclusion chromatography. PMID- 7496483 TI - Selection of peptides with surface affinity for alpha-chymotrypsin using a phage display library. AB - Peptides with affinity for the surface of alpha-chymotrypsin (EC 3.4.21.1) were selected from a hexapeptide phage display library consisting of approximately 10(7) different clones. Seven selections were performed and five individual phage clones analysed. Compared to the primary library, the five peptide phage clones all interacted more strongly with alpha-chymotrypsin, and DNA sequencing of the phage clones revealed five different amino acid sequences: Gly-Ala-Val-Ile-Thr His, Arg-Asp-Ile-Val-Val-Ala, Val-Tyr-Ser-His-Ala-Ser, Gly-Ser-Tyr-Ser-Ala-Gly and Leu-Asp-Ile-Val-Val-Ala. Two of the peptides exhibited 83% identity (i.e. a difference of just one amino acid). The chemically synthesized peptides competitively reduced the binding of the corresponding peptide phage clone to alpha-chymotrypsin. Binding of some of the selected peptide phage clones to alpha chymotrypsin was also reduced by several of the other non-corresponding synthesized peptides, suggesting that these peptides have common recognition areas on the enzyme. Three of the synthesized peptides were poor substrates of alpha-chymotrypsin and they did not inhibit enzyme activity. Our results suggest that it is possible to select peptides from peptide phage display libraries with affinity for different surface structures on the enzyme, not involved in the biologically active site. PMID- 7496484 TI - Purification of recombinant human basic fibroblast growth factor: stability of selective sorbents under cleaning in place conditions. AB - Human basic fibroblast growth factor (bFGF) was produced from recombinant Escherichia coli by high-cell-density cultivation. In order to develop a purification strategy for large-scale purification, chromatographic sorbents with different anionic functional groups were compared in terms of selectivity for bFGF and stability under cleaning in place (CIP) conditions. Heparin-Sepharose CL 6B, Fractogel EMD-SO3- 650 (S) and SP-Sepharose (high performance) were found suitable for this purpose with decreasing selectivity in that order. Each sorbent was treated eight times under CIP conditions employing both 0.2 and 1.0 M NaOH, in order to study modifications of these sorbents. Heparin-Sepharose displayed more than 50% loss of capacity after the first CIP treatment and decreasing selectivity with each cycle. Both cation exchangers displayed almost constant results regarding selectivity and capacity. The Fractogel EMD-SO3- exhibited only slightly lower selectivity for bFGF than Heparin-Sepharose and the highest capacity of all sorbents tested. Agglomeration of bFGF at low salt concentrations was a serious problem. By direct application of pooled fractions from Fractogel EMD-SO3- onto Heparin-Sepharose a highly pure product was obtained; however, the recovery after Heparin-Sepharose was only 30%. PMID- 7496485 TI - Identification of a new genetic variant of bovine beta-casein using reversed phase high-performance liquid chromatography and mass spectrometric analysis. AB - Various components of the beta-casein fraction from bovine milk were separated by preparative reversed-phase high-performance liquid chromatography (RP-HPLC). They included the genetic variants beta A1, beta A2, beta A3, and an unknown component previously denoted beta X [S. Visser et al., J. Chromatogr. 548 (1991) 361-370]. Tryptic digests of these components were compared by RP-HPLC and most peaks were analysed by mass spectrometry (MS). The tryptic map of beta X was closest to that of beta A1, but with a few mutually different peak components. Electrospray ionisation MS revealed that in the beta X map these components had relative molecular masses of 16 higher than the corresponding ones in the beta A1 map. The main differential peaks represented the 114-169 fragments of beta A1 and beta X, respectively, which were both purified and then cleaved with cyanogen bromide. In the resulting mixtures, each of which contained three fragments, the corresponding peptides representing the 145-156 sequence showed the 16 relative molecular mass difference. In beta X this sequence contained a Leu residue at position 152 instead of the Pro-152 in beta A1, as established by fast-atom bombardment MS-MS. The Leu could be discriminated from an Ile residue by the presence of a side-chain-specific, D-type fragment ion in the MS-MS spectrum of the beta X CNBr peptide. The sequence of the two homologous 145-156 fragments was confirmed by regular amino acid sequence analysis. In accordance with internationally accepted guidelines for the nomenclature of milk proteins, the new genetic variant has been named beta-casein F-5P. PMID- 7496486 TI - Purification of monoclonal antibodies on dextran-coated silica support grafted by thiophilic ligand. AB - Coated silica beads are promising supports for high-performance liquid chromatography (HPLC) of proteins; they combine the excellent mechanical properties of silica with minimal non-specific interactions with proteins in solution due to the presence of hydrophilic dextran polymers adsorbed at the silica surface. So, dextran-coated porous silica beads can be grafted with beta mercaptoethanol by using divinylsulfone as coupling reagent to obtain new thiophilic supports usable in HPLC. The affinity of monoclonal IgG subclasses from mouse ascitic fluids for the active phases can be analysed. These dextran coated silica supports grafted with thiophilic ligands allow a one-step purification of these antibodies. Moreover, the chromatographic separation of two subclasses, immunoglobulin G1 (IgG1) and IgG3, is observed and can be correlated to the high resolution of these new HPLC thiophilic supports. PMID- 7496487 TI - Separation and characterisation of bovine histone H1 subtypes by combined ion exchange and reversed-phase chromatography and mass spectrometry. AB - In order to separate and identify histone H1 subtypes from calf thymus we used both electrospray mass spectrometry (ES-MS) and matrix-assisted laser desorption ionisation time-of-flight mass spectrometry (MALDI-TOF-MS) after a three-step chromatographic procedure consisting of reversed-phase high-performance liquid chromatography (RP-HPLC), size-exclusion chromatography (SEC) and ion-exchange chromatography (IEC). Under the RP-HPLC conditions described, we obtained two baseline-separated H1-fractions which were characterised by MALDI-TOF-MS. The determined masses ranged from 22,850 to 22,590 for the first fraction and from 22,070 to 21,250 for the second fraction. Further, it was shown that the first fraction contained at least four and the second one at least five subtypes of the histone class H1. Four homogeneous pure H1 subtypes were obtained by a combination of IEC followed by SEC and RP-HPLC. The molecular masses of these four subtypes determined by ES-MS were 22,606, 22,761, 21,347 and 21,263. We obtained six additional molecular masses of histone H1 subtypes from three heterogeneous fractions, namely 22,066, 21,802, 20,586 and 19,817 by ES-MS and 22,800 and 22,675 by MALDI-TOF-MS. The retention times of these fractions and the molecular masses were in agreement with the data obtained from RP-HPLC fractions by MALDI-TOF-MS. PMID- 7496488 TI - Effect of glycosylation on the heparin-binding capability of boar and stallion seminal plasma proteins. AB - Boar and stallion seminal plasmas were fractionated using affinity chromatography on heparin-Sepharose. In both species, among other proteins, the heparin-binding (H+) and non-heparin-binding (H-) fractions each contained glycoforms of either porcine PSP-I or equine HSP-1 and HSP-2. However, porcine H+/PSP-I eluted as a monomeric protein, whereas H-/PSP-I formed a heterodimer with PSP-II, another major seminal plasma protein. On the other hand, the stallion proteins H+/HSP-1 and H+/HSP-2 eluted together as an aggregate of relative molecular mass (M(r)) 90,000, whereas H-/HSP-1 and H-/HSP-2 eluted as monomers (15,000). Remarkably, when PSP-I and PSP-II from the H- fraction were separated, both proteins bound to heparin. Altogether these data show that glycosylation has an indirect effect on the heparin-binding ability of PSP-I, HSP-1 and HSP-2 through modulation of their aggregation state. PMID- 7496489 TI - High-performance liquid chromatographic purification of extremely hydrophobic peptides: transmembrane segments. AB - Transmembrane peptides of integral membrane proteins often exhibit extremely high hydrophobicity. Therefore, the solubility of such peptides in solvents commonly used in HPLC is usually very low and the interaction with generally applied stationary phases such as silica gel or C18 reversed phases appears to be extremely strong, which makes the characterization and purification of these peptides difficult. The analytical characterization and preparative separation of the synthesized M1 transmembrane sequence of the inhibitory glycine receptor M(r) 48,000 subunit and some of its fragments is shown. M1 and its larger fragments could be dissolved in a dichloromethane-hexafluoro-2-propanol mixture containing a trace amount of pyridine for their separation on a C4 phase by employing linear two-component gradients of formic acid-2-propanol and formic acid-water with ratios up to 4:1 (v/v). Conditions to avoid formylation of the peptides are indicated. PMID- 7496490 TI - Characterization of molecular aggregates of alpha 1 beta 1-integrin and other rat liver membrane proteins by combination of size-exclusion chromatography and chemical cross-linking. AB - Many membrane proteins display their biological activity in molecular aggregates of interacting counterparts. The analysis of these aggregates remains difficult; especially intermolecular complexes of membrane proteins tend to dissociate or artificially aggregate during detergent extraction out of membranes. Thus, the existence of protein aggregates was investigated by two approaches. First, after modest detergent extraction, the presence of three well characterized rat liver membrane proteins, alpha 1 beta 1-integrin, dipeptidyl aminopeptidase IV (DPP IV) and cell-CAM 105 (CAM = cell adhesion molecule), in aggregates could be demonstrated when investigated by size-exclusion chromatography (SEC) under non denaturating conditions. However, the applied detergents partially influenced the resolution of the separation reducing the ability to discriminate between native and artificial protein aggregates. To circumvent these problems, a second approach based on covalent cross-linking of native protein complexes by dithiobis(succinimidylpropionate) was combined with the performance of denaturating SEC. Under such optimized some high-molecular-mass complexes of all model proteins consisting of unknown components could also be detected. Taken together, non-denaturating SEC and chemical cross-linking in combination with denaturating SEC represent methodological approaches for the characterization of protein aggregates. PMID- 7496491 TI - Liquid chromatographic determination of D- and L-amino acids by derivatization with o-phthaldialdehyde and N-isobutyryl-L-cysteine. Applications with reference to the analysis of peptidic antibiotics, toxins, drugs and pharmaceutically used amino acids. AB - In order to evaluate and extend the applicability of an analytical method that enables the quantitative and simultaneous high-performance liquid chromatographic determination of D- and L-amino acids (DL-AAs) by automated precolumn derivatization with o-phthaldialdehyde together with the chiral thiol N isobutyryl-L-cysteine [J. Chromatogr., 666 (1994) 259] selected natural and synthetic bioactive peptides, as well as pharmaceutically used formulations of AA, were investigated and the amounts of D- and L-AA determined by fluorescence detection. Peptides containing cys(e)ine were oxidized with performic acid prior to hydrolysis with 6 M HCl, and those containing Trp were hydrolyzed with 4 M methanesulfonic acid (24 h at 110 degrees C in both cases). Peptides analyzed were the peptide antibiotics bacitracin, gramicidins A and S, polymyxin B, metanicin C, the peptide toxin malformin A and the peptide drugs D-Arg [Hyp3,Thi5,8,D-Phe7]-bradykinin, beta-casomorphin and alpha s1-exorphin. Further, the enantiomeric ratios of pharmaceutically used AA formulations containing racemic DL-Ser, DL-Asp and DL-Met were determined, and the AA drugs L-Asp and L Trp were tested negatively for the presence of the respective D-enantiomers. In two aqueous formulations of L-AA used for parenteral nutrition, low amounts of D AA (0.1-0.9% with respect to certain L-AA enantiomers and of totally 128 mg and 149 mg D-AAs per liter infusion solution) were determined. PMID- 7496493 TI - Perfusion immunoassay for acetylcholinesterase: analyte detection based on intrinsic activity. AB - A quantitative, two-column, HPLC-based assay requiring only 30 min to complete is reported. Amniotic fluid proteins are first fractionated on a size-exclusion column; the fraction containing the M(r) 280,000 neural acetylcholinesterase (AChE) is then diverted to a second column, an ImmunoDetection cartridge derivatized with an anti-AChE antibody. The immobilized antibody traps the enzyme, then substrate is flowed through the cartridge and the product is detected. For a positive result, the enzyme must have a molecular mass corresponding to the neural-AChE, be recognized by the antibody and be active in converting the substrate into product. The assay is sensitive in the clinically relevant range. The method provides rapid quantitative analysis using an automated instrument projected to be suitable for screening large numbers of samples. PMID- 7496492 TI - Capillary and rotating-tube isoelectric focusing of a transmembrane protein, the human red cell glucose transporter. AB - The human red cell glucose transporter (Glut1) is a transmembrane protein. Monomeric Glut1 was purified by ion-exchange chromatography in the presence of the non-ionic detergent n-dodecyl octaoxyethylene (C12E8). For focusing, the ionic strength of the solution of C12E8-Glut1 complexes with co-purified lipids was lowered by dialysis, the detergent concentration was increased and carrier ampholytes were added. Focusing was done for 5 min at 3000 V in a methyl cellulose-coated glass capillary (50 microns I.D.). The anolyte H3PO4 was then replaced by NaOH for mobilization towards the anode. Absorbance monitoring at 280 nm showed two groups of zones at pH 6 and 8. Similarly, isoelectric focusing in a rotating quartz tube (3 mm I.D.) gave Glut1 zones at pH 5.5 and 8.0. Phosphorus analysis revealed that the Glut1 zone at pH 8 contained more phospholipids than did the other one. The above results together with previously determined and calculated isoelectric points (pI) of Glut1 indicate that the Glut1 at pH 8 is monomeric and that the zone at pH 5.5-6 represents oligomeric materials. The pI 8.0 at 22 degrees C applies for monomeric Glut1 in the absence of urea. The results exemplify that capillary isoelectric focusing of hydrophobic membrane proteins is possible. PMID- 7496494 TI - Novel magnetic microspheres on the basis of poly(vinyl alcohol) as affinity medium for quantitative detection of glycated haemoglobin. AB - A water-in-oil suspension cross-linking technique using poly(vinyl alcohol) as polymer phase to prepare novel magnetic microbeads is described. By dispersing a conventional Fe3O4 pigment in the polymer phase and subsequently suspending the mixture in a vegetable oil phase with a defined viscosity, spherical magnetic microspheres are obtained. Bead sizes ranging from 1 to 50 microns and above can be obtained by exploiting well defined preparation parameters such as polymer concentration and oil and polymer viscosity. The performance of the magnetic matrices for the separation and quantification of glycated haemoglobin was tested using a m-aminophenylboronic acid matrix. The feasibility of this detection method for blood sugar diagnosis is discussed using a commercial column test kit for comparison. PMID- 7496495 TI - Conformational effects in reversed-phase high-performance liquid chromatography of polypeptides. I. Resolution of insulin variants. AB - In order to further characterise the role of conformation in the retention behaviour of polypeptides and proteins in reversed-phase high-performance liquid chromatography (RP-HPLC), the chromatographic properties of four different insulins have been studied as a function of temperature (over the range 5-85 degrees C) and column residence time (over the range 10-60 min). The role of the ligand structure was also investigated by comparing results obtained with a n octadecyl (C18) and a n-butyl (C4) ligand immobilised to the same porous silica. Comparative structure-retention-stability relationships were determined from an examination of the influence of temperature on a number of chromatographic parameters including the chromatographic contact area, the affinity constant and the experimental band width. The results demonstrated that variations in temperature can be used to affect significant changes in selectivity between the different insulins despite their very high degree of sequence homology. These observations have permitted specific amino acid residues, and in particular those residues encompassing the region A8-A10, to be proposed to be directly involved in the chromatographic contact area of the insulin molecules. Overall, the analysis of the changes in various chromatographic parameters in response to variation of the amino acid sequence, temperature and other experimental parameters provides a powerful tool to elucidate the structural basis for the interfacial stability and the role of conformation on the retention behaviour of polypeptides and proteins in RP-HPLC. PMID- 7496496 TI - Conformational effects in reversed-phase high-performance liquid chromatography of polypeptides. II. The role of insulin A and B chains in the chromatographic behaviour of insulin. AB - The contribution of the insulin A- and B-chain to the retention and bandwidth behaviour of bovine insulin has been investigated. The influence of temperature and residence time on the logarithmic capacity factor (log k) versus the mole fraction of organic modifier psi, i.e. the effect of temperature and ligand residency on the S and log k0 values of the individual peptide chains, were assessed at temperatures between 5 and 85 degrees C and elution times between 30 to 90 min with an n-octadecyl (C18) and an n-butyl (C4) sorbent. Analysis of these log k versus psi dependencies revealed that the insulin A-chain exhibits retention behaviour significantly different to the intact insulin molecule whilst the B-chain exhibits retention behaviour which is remarkably similar to the parent protein. However, in terms of kinetic processes, the A-chain exhibited a peak-splitting phenomenon at higher temperatures which was similar to the behaviour of the intact insulin molecule, whilst only bandbroadening with no peak splitting was apparent for the B-chain. Overall, the similarity of the retention behaviour of the insulin B-chain and the intact insulin molecule with regard to their temperature and residency dependencies suggests that the insulin B-chain makes a significant contribution to the chromatographic contact region of the insulin molecule when this polypeptide is exposed to hydrocarbonaceous ligands at low to intermediate temperatures due to the progressive unfolding of the molecule and greater accessibility of the previously buried B-chain residues.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7496498 TI - Interaction mechanisms between insulin and N-acetylneuraminic acid in affinity chromatography. AB - Silica beads are coated with dextran carrying a calculated amount of positively charged diethylaminoethyl (DEAE) groups in order to neutralize negatively charged silanol groups at the silica surface and in this way to minimize non-specific interactions between silica and proteins in solution. Dextran-coated silica supports are potentially excellent stationary phases for high-performance liquid chromatography of proteins. These supports combine the advantages of polysaccharide phases with the excellent mechanical characteristics of silica. These supports [silica-dextran-DEAE (SID)] are easily functionalized by grafting N-acetylneuramic acid (NANA), extracted from edible birds' nests, using conventional coupling methods. The performance of supports bearing NANA was studied by high-performance liquid affinity chromatography of insulin, the hypoglycaemic peptide hormone of the human organism. The study showed that these supports exhibit a reversible and specific affinity towards insulin and allow separations with high purification yields. The influence of different physico chemical parameters (pH, temperature and insulin concentration) on insulin retention on the support was studied. This allowed the optimization of the conditions of adsorption and a better understanding of the interaction mechanisms between insulin and NANA as a biospecific ligand. PMID- 7496499 TI - Mucosa-associated lymphoid tissue (MALT) lymphoma. PMID- 7496497 TI - Removal of endotoxins by affinity sorbents. AB - Histidine, histamine and polymyxin B affinity sorbents were employed for the removal of Escherichia coli-derived endotoxins. Their effectiveness was compared with those of poly-L-lysine-Sepharose and DEAE-Sepharose. All sorbents reduced the concentration of endotoxins from an E. coli culture filtrate to tolerable levels. However, their effectiveness was not higher than that of the anion exchanger, which displayed clearance rates of up to 15,000. Endotoxin removal from protein solutions depended on the net charge of the desired protein. Lysozyme as a model for positively charged proteins enhanced endotoxin removal. In contrast, only low initial contamination levels (< 34 EU/ml) were reduced to tolerable levels from bovine serum albumin (BSA) as the negatively charged protein model owing to competition of BSA and endotoxins for adsorption sites. Hence also a low BSA recovery was observed after the treatment whereas the lysozyme recovery was almost 100%. At pH values below the isoelectric point of BSA, endotoxin removal was also more effective. The best conditions for the decontamination were found at neutral pH and low ionic strength (< or = 20 mM phosphate). Ionic forces between ligands and endotoxins are dominant at this ionic strength; hydrophobic interactions are not very effective. Hence the selectivities of all sorbents towards endotoxins are not exceptionally high. DEAE anion exchangers are the most suitable sorbents for the removal of endotoxins from solutions accommodating positively charged proteins owing to their low cost and high capacity. Poly-L-lysine-Sepharose was most effective for the removal of small amounts of endotoxins from solutions of negatively charged proteins. The "affinity ligands" histamine, histidine and polymyxin B were effective for the removal of endotoxins from E. coli filtrate; however, their effectiveness decreased dramatically in the presence of BSA and it was lower than for poly-L lysine- and DEAE-Sepharose in the presence of lysozyme. PMID- 7496500 TI - Evolution of three patterns of intra-alveolar fibrosis produced by bleomycin in rats. AB - In pulmonary fibrosis, it is known that fibrotic changes develop in the intra alveolar spaces and that intra-alveolar fibrosis can be classified into three patterns, namely intra-alveolar buds, mural incorporation and obliterative changes. In order to clarify the evolution of intra-alveolar fibrosis, immunohistochemical studies of extracellular matrix proteins and electron microscopic observations were made of the lungs of rats given a single intratracheal instillation of bleomycin. All three patterns of fibrosis developed in this model. Intra-alveolar buds changed into globular lesions with dense collagen deposition, the surface of which was covered by alveolar epithelium. Electron microscopy revealed that the buds often contained spiraling collagen fibrils and numerous microfibrils, but not mature elastic fibres, beneath the regenerating epithelial lining cells; the epithelial basement membranes were discontinuous. In contrast, mural incorporation and obliterative changes were associated with alveolar structural remodeling. Electron microscopically, these lesions had bundles of normal collagen fibrils, small elastic fibers, and continuous epithelial basement membranes. These results indicate that: (i) intra alveolar buds, that become intra-alveolar collagen globules, with an unusual extracellular matrix, do not contribute to alveolar structural remodelling; and (ii) areas of mural incorporation and obliterative changes have the usual type of extracellular matrix and are essential for alveolar structural remodelling. PMID- 7496501 TI - Intercellular adhesion molecule-1 expression by macrophages in human gastrointestinal carcinoma: possible roles as host immune/inflammatory reaction. AB - Tumor-associated macrophages (TAM) are one of the factors which modulate the carcinoma progression. The present study described immunohistochemical expression of intercellular adhesion molecule-1 (ICAM-1) in stromal cells in human gastrointestinal carcinoma identifying the cell types by immunoelectron microscopy. In colon and gastric carcinomas, ICAM-1-positive cells were mostly stromal cells, and major cell types were identified as macrophages and fibroblasts by immunoelectron microscopy. Macrophages were characterized by their ovoid shape, cytoplasmic projections, abundant vacuoles, phagocytosis, and paucity of rough endoplasmic reticulum. Fibroblasts contained stacks of rough endoplasmic reticulum. Macrophages were major cells among ICAM-1-positive cells along the invasive margin, while fibroblasts were predominant in the stroma within carcinoma in colon and intestinal-type gastric carcinomas. Lymphocytes positive for lymphocyte function associated antigen (LFA-1), a counter-receptor of ICAM-1, were densely distributed along the invasive margin, and sparsely in the stroma within carcinoma. In diffuse-type gastric carcinoma, most macrophages were dendritic-shaped and negative for ICAM-1. Our study suggests that the invasive margin is an area similar to active inflammation, where the antigen presenting cells (macrophages) and lymphocytes may interact via the ICAM-1/LFA-1 adhesion. PMID- 7496502 TI - Morule with biotin-containing intranuclear inclusions in thyroid carcinoma. AB - One thousand and sixty cases of thyroid carcinoma were reviewed to compare morules with squamous metaplasia clinicopathologically and immunohistochemically. Morules and squamous metaplasia were found in five (0.47%) and 32 cases (3.0%) respectively. The five patients with morules were all female (age 20-36 years) including four with papillary carcinoma and one with follicular carcinoma. The 32 patients with squamous metaplasia consisted of 30 females and 2 males (age 14-78 years), all of whom had papillary carcinoma except for one follicular carcinoma. The morules demonstrated characteristic 'optically clear nuclei' (OCN), which ultrastructurally showed filamentous structures in the nuclei. The OCN were immunohistochemically demonstrated to contain intranuclear biotin. Furthermore, the morule often accompanied with the OCN was positive for Ulex Europaeus agglutinin l (UEA-l) but negative for bovine muzzle epidermal keratin (EK). On the contrary, squamous metaplasia unaccompanied with the OCN was negative for UEA l, but positive for EK. Follow-up information revealed that one of the five patients with morules had died of the disease, one was alive with pulmonary metastasis, and three were disease-free. Eight of 32 patients with squamous metaplasia had died of the disease; of the others who were alive, four patients have had recurrence. PMID- 7496504 TI - Spindle cell hemangioendothelioma: an immunohistochemical and flow cytometric study of six cases. AB - Six cases of spindle cell hemangioendothelioma (SCH) are presented with immunohistochemical and flow cytometric analyses. One case was associated with Maffucci's syndrome. All lesions were solitary or multifocal in the extremities, and prepresentation duration ranged from years to decades. One case recurred but none had metastases. Histologically, in four of the six cases the main lesions appeared to arise within vessels, predominantly muscular vessels. All lesions consisted of cavernous hemangioma-like areas and solid cellular areas resembling Kaposi's sarcoma. Cellular atypia was minimal. At the periphery of the lesions, a cluster of large thick or thin walled, and probably malformed, vessels were observed. Immunohistochemically, factor-VIII related antigen, CD34, vimentin, and lectin binding Ulex europaeus agglutinin 1 stained endothelial cells lining vascular channels, and vacuolated, or epithelioid cells. Spindle cells in the solid areas were negative for these endothelial markers except for vimentin, but showed divergent positive immunoreactions of HHF35, alpha-smooth muscle actin, desmin, and collagen type IV. Five cases were diploid and one was aneuploid. There was no significant correlation among DNA ploidy, S-phase fraction, and local recurrence in SCH although the number of cases examined was small. These results suggest SCH may be a benign lesion, probably a reactive process, rather than a low-grade angiosarcoma. PMID- 7496503 TI - Six cases of microscopic polyarteritis exhibiting acute interstitial pneumonia. AB - Out of 21 autopsy cases of microscopic polyarteritis (MPA), six cases that died of acute respiratory failure are described. All of these six cases exhibited pauci-immune necrotizing glomerulonephritis. Of these six cases, systemic vasculitis and respiratory failure occurred almost simultaneously in two, whereas respiratory failure developed when vasculitis recurred in the other four. Pathologically, pulmonary changes were diffuse and almost uniform in each case. Interstitial inflammatory cells (alveolitis), hyaline membrane, restructuring, and fibrosis were observed. The degrees of these changes differed from case to case; four cases showed predominantly exudative changes and two cases exhibited proliferative or organizing ones. Clinical and pathological features were consistent with acute interstitial pneumonia (AIP). Immunofluorescent and ultrastructural studies did not suggest deposition of immune complexes at the lesions of alveolitis. An etiologic agent could not be identified in these cases. Some lung lesions including pulmonary hemorrhage have been described in association with MPA; however, cases such as those presented here have rarely been reported. It was considered that AIP could be one of the pulmonary disorders in MPA. Furthermore, the possible pathogenetic role of antineutrophil cytoplasmic antibodies (ANCA) in the evolution of present lung lesion is discussed. Although ANCA were not examined in the present cases, these antibodies should be investigated in future cases. PMID- 7496505 TI - Immunohistochemical study on collagenous proteins and biophysical analysis of crystals in extraskeletal chondroma. AB - The immunohistological distribution of collagen types I, II, III, and VI in five cases of extraskeletal chondroma was examined and compared with that in six cases of enchondroma. In addition, the composition of crystals deposited in three cases of extraskeletal chondroma were biophysically analyzed with special attention to the relationship between the collagen types of the matrix and the crystal deposition. In extraskeletal chondroma, immunoreactivity of type II collagen in the extracellular matrix and type VI collagen in the pericellular area, which were strongly and diffusely recognized in the normal hyaline cartilage and enchondroma, was diminished. Instead, additional types of collagen, types I and III, were demonstrated in the matrix. Electron roentgenographic microanalysis and infrared light spectroscopic analysis revealed that calcium pyrophosphate dihydrate (CPPD) was included in the crystals of extraskeletal chondroma. CPPD crystals were observed in/around collagen types I and III. The possible relationship between the difference of collagen composition in the matrix and the CPPD crystal deposition is discussed. PMID- 7496506 TI - Monoclonality of normal human colonic crypts. AB - The monoclonality of human colonic crypts was demonstrated by human androgen receptor (HUMARA) gene assay following application of the crypt isolation method. DNA was extracted from an isolated single crypt, Hpa II digestion was performed before polymerase chain reaction (PCR) by primers spanning the HUMARA exon 1 region. The PCR product of a single crypt clearly showed allelic exclusion based on methylation status, while PCR product from a mixture of 40 crypts or colonic mucosa as a whole that included epitheliums and interstitial connective tissue had two bands. This method will facilitate the non-isotopic analysis not only of tumor clonality, but also of the normal structures derived from a single progenitor cell. PMID- 7496507 TI - Small cell neuroendocrine carcinoma of the rectum. AB - Poorly differentiated small cell neuroendocrine (NE) carcinoma of the colon and rectum is a rare primary epithelial malignancy at this location. A case of a highly aggressive NE tumor of small cell type, combined with non-invasive well differentiated papillary adenocarcinoma in villous adenoma is reported. The patient died rapidly with massive and progressive liver metastasis. The tumor cells were argyrophilic and diffusely immunoreactive for neuronspecific enolase and synaptophysin. Ultrastructural analysis disclosed NE-type cored granules in most of the small tumor cells. NE tumors of the colon and rectum are briefly reviewed. PMID- 7496508 TI - Minigemistocytic astrocytoma with frequent apoptoses: analysis of tumor growth. AB - A rare glial tumor known as 'minigemistocytic astrocytoma (gliofibrillary oligodendroglioma)' is reported in a 73 year old Japanese male. A low-density area found by computed tomography and thought to be an operative scare remaining after hematoma in the right frontal lobe of the cerebrum had been followed for 10 years. This area, however, had been accompanied by a cyst for 2 years and had developed gradually for 1 year prior to dissection. The tumor was poorly demarcated from the surrounding normal tissue macroscopically at operation. Microscopically, the tumor consisted of small gemistocytic cells in uniform sheets intersected by a small vascular stroma with frequent eosinophilic granular bodies, mitoses and apoptotic bodies. Immunohistochemical examination for glial fibrillary acidic protein (GFAP) revealed remarkable positive reactivity in the perinuclear cytoplasm, but no immunoreactivity for vimentin or Leu 7 was found. Electron microscopically, rich filaments arranged in parallel bundles were found in the neoplastic cells. These histological findings are closely consistent with those of previously reported minigemistocytic astrocytoma cases. The GFAP-rich minigemistocytic astrocytoma with granular bodies and frequent mitoses in the present case is considered to indicate a higher degree of astrocytic differentiation and malignant potential than previous cases. The frequent apoptoses, however, might inhibit tumor growth in this case. PMID- 7496510 TI - Pericardial malignant mesothelioma: case report and discussion of immunohistochemical and histochemical findings. AB - An extremely rare case of malignant mesothelioma of the pericardium is reported. The tumor appeared to be the biphasic type of diffuse malignant mesothelioma which present many small to fine lipid droplets in the cytoplasm, particularly that of the epithelial cell, and a positive immunohistochemical reaction with antibodies to low molecular weight cytokeratin (45-50 kDa) in all of the tumor cells. The literature is also reviewed and the presence of lipid droplets and the importance of the immunohistochemical positivity to low molecular weight cytokeratin for differential diagnosis from other malignant tumors, such as invasive adenocarcinoma or primary sarcoma of the pleura, or soft tissue sarcoma involving the pleura is discussed. PMID- 7496509 TI - Intracranial meningioma masquerading as a primary pleuropulmonary tumor. AB - Extracranial metastasis was found in an intracranial meningioma with low proliferative potential before the detection of the primary tumor. Pleuropulmonary tumors were incidentally detected on chest X-ray in an asymptomatic 25 year old female. Excised tumors of the right pleura and lung showed histological features similar to meningotheliomatous meningioma, which led to the discovery and excision of the intracranial tumor. Both tumors showed the same histologic pattern: meningotheliomatous meningioma with low mitotic activity. The proliferative component, determined by the monoclonal antibody Ki 67, was further evaluated in the primary tumor and the metastases of the present case, as well as in 12 other intracranial meningiomas. Ki-67 positive ratios at the primary and metastatic sites of the present cases were 1.2 and 1.1%, respectively, which is as low as other benign meningiomas, and this suggests that factors other than the proliferative potential is responsible for extracranial metastasis of meningioma. PMID- 7496511 TI - Soft X-rays--a novel technique for the study of bacteria. PMID- 7496512 TI - Bacterial polysaccharides in sickness and in health. The 1995 Fleming Lecture. PMID- 7496513 TI - alpha-D-glucuronidases from the xylanolytic thermophiles Clostridium stercorarium and Thermoanaerobacterium saccharolyticum. AB - alpha-D-Glucuronidases were purified from the xylanolytic thermophiles Clostridium stercorarium and Thermoanaerobacterium saccharolyticum. This enzyme activity was found to be intracellular in each organism, with T. saccharolyticum producing much greater total activity. The specific activities of the purified enzymes (10 U mg-1 T. saccharolyticum; 1.7 U mg-1 C. stercorarium) differed by a factor of approximately 5. For the determination of enzyme activities, 4-O-methyl alpha-D-glucuronosyl-xylotriose was used as a substrate and the glucuronic acid released by alpha-D-glucuronidase action was quantified by a colorimetric procedure. 4-O-Methyl-alpha-D-glucuronosyl-xylotriose was the hydrolysis product that accumulated after exhaustive degradation of 4-O-methyl-alpha-D glucuronoxylan with xylanases of C. stercorarium. Hydrolysis of side chains in high-molecular-mass glucuronoxylan could not be detected. Neither of the enzymes was able to hydrolyse the chromogenic aryl-substrate p-nitrophenyl-alpha-D glucuronoside. Both alpha-D-glucuronidases have a dimeric structure, with monomeric molecular masses of 72 and 76 kDa for C. stercorarium and of 71 kDa for T. saccharolyticum. The pI was estimated to be 4.3 for each enzyme. While both enzymes exhibited a similar pH optimum (pH 5.5-6.5) they differed in their thermostabilities. At 60 degrees C, half-lives of 14 and 2.5 h, respectively, were determined for the alpha-D-glucuronidases of C. stercorarium and T. saccharolyticum. This description of alpha-D-glucuronidase activity in thermophilic anaerobic bacteria extends our knowledge of these enzymes, previously purified and characterized only in fungi. PMID- 7496514 TI - Distribution of the smpA gene from Serpulina hyodysenteriae among intestinal spirochaetes. AB - Forty intestinal spirochaete strains were investigated for nucleotide sequences related to the smpA locus from Serpulina hyodysenteriae by Southern hybridization of chromosomal DNA using the smpA locus from S. hyodysenteriae strain P18A as a probe and by PCR using primers internal to the smpA gene. The intensity of the hybridization signal at high stringency and positive PCR results suggested that 12 S. hyodysenteriae strains possessed a similar nucleotide sequence. PCR was negative for another 12 S. hyodysenteriae strains and the hybridization signal obtained from 11 of these was weak and one was negative. All S. hyodysenteriae strains hybridized under low stringency conditions. These results indicated that there is variation among the smpA loci of S. hyodysenteriae strains. Among seven strains of S. innocens, and the proposed species 'S. intermedius' and 'S. murdochii', hybridization was weak and no PCR products were obtained, suggesting that these species have sequences related to, but divergent from, the smpA sequences of strains of S. hyodysenteriae. Both gene probe hybridization and PCR analysis of nine strains of the proposed new genus 'Anguillina', including isolates from pigs and humans, gave negative results. PMID- 7496515 TI - A murC gene in Porphyromonas gingivalis 381. AB - The gene encoding a 51 kDa polypeptide of Porphyromonas gingivalis 381 was isolated by immunoblotting using an antiserum raised against P. gingivalis alkaline phosphatase. DNA sequence analysis of a 2.5 kb DNA fragment containing a gene encoding the 51 kDa protein revealed one complete and two incomplete ORFs. Database searches using the FASTA program revealed significant homology between the P. gingivalis 51 kDa protein and the MurC protein of Escherichia coli, which functions in peptidoglycan synthesis. The cloned 51 kDa protein encoded a functional product that complemented an E. coli murC mutant. Moreover, the ORF just upstream of murC coded for a protein that was 31% homologous with the E. coli MurG protein. The ORF just downstream of murC coded for a protein that was 17% homologous with the Streptococcus pneumoniae penicillin-binding protein 2B (PBP2B), which functions in peptidoglycan synthesis and is responsible for antibiotic resistance. These results suggest that P. gingivalis contains a homologue of the E. coli peptidoglycan synthesis gene murC and indicate the possibility of a cluster of genes responsible for cell division and cell growth, as in the E. coli mra region. PMID- 7496517 TI - L-arginine transport and metabolism in Giardia intestinalis support its position as a transition between the prokaryotic and eukaryotic kingdoms. AB - Arginine is metabolized by the arginine dihydrolase pathway in Giardia intestinalis trophozoites and is an important metabolic fuel for this parasite. Radiolabelled arginine was used to characterize the transport of arginine into Giardia intestinalis trophozoites. The transporter had a high affinity for arginine (Km 15 microM) and a high activity [Vmax 76 nmol min-1 (mg protein)-1 at 25 degrees C]. Substrate specificity studies indicated an absolute requirement for the alpha-amino and carboxyl groups, but a tolerance for some substitutions in the guanidino group. The use of non-metabolized arginine analogues in combination with HPLC amino acid analysis of intra- and extracellular pools demonstrated that the arginine transporter is an arginine-ornithine antiport. Investigations of the first step of arginine metabolism, involving arginine deiminase, revealed a relatively high affinity for arginine (Km 0.16 mM) and a large maximal velocity [Vmax 550 nmol min-1 (mg protein)-1 at 37 degrees C]. Substrate specificity studies showed that the arginine deiminase had a characteristically different substrate recognition profile to that of the arginine transporter. Overall, the combination of the transporter and the deiminase result in very low intracellular arginine concentrations and their properties are consistent with the rapid transport of arginine for metabolism via the arginine dihydrolase pathway. PMID- 7496516 TI - Listeria monocytogenes exists in at least three evolutionary lines: evidence from flagellin, invasive associated protein and listeriolysin O genes. AB - Regions of the genes encoding flagellin (flaA), the invasive associated protein (iap), listeriolysin O (hly) and 23S rRNA were sequenced for a range of Listeria monocytogenes isolates of different origin and serotypes. Several nucleotide sequence variations were found in the flaA, iap and hly genes. No differences were found for the rRNA genes, but our approach does not exclude the existence of differences between single copies of these genes. Based on the sequence differences, the L. monocytogenes strains can be divided into three distinct sequence types. Further, the presence of only a small number of sequence differences within each group indicates a strong degree of conservation within the groups. There was a complete correspondence among the groups of strains formed according to the analysis of the flaA, iap and hly genes, and the grouping correlates with serotype, pulsed field gel electrophoretic and multilocus enzyme electrophoretic data. Analysis of the region encoding the threonine-asparagine repeat units in the iap gene revealed some striking features. Sequence type 1 strains were found to have 16-17 repeats, sequence type 2 strains had 16-20 repeats whereas the two sequence type 3 strains analysed had only 11 repeats. Furthermore, within a 19 bp segment there was a 37% difference between the sequences of type 1 and 2 strains and that segment was absent in type 3 strains. Within the threonine-asparagine repeat region the nucleotide differences gave rise to four amino acid changes; however, all were changes among the three amino acids present in the repeat structure indicating a strong selective pressure on the composition of this region. PMID- 7496518 TI - Expression of foreign genes and selection of promoter sequences in Acholeplasma laidlawii. AB - The stable maintenance and expression of foreign genes in mollicutes (mycoplasmas) have been difficult to achieve due to the lack of suitable vectors. In this paper we show for the first time that a replicating vector can been used to express foreign genes other than antibiotic resistance genes in Acholeplasma laidlawii. Plasmids derived from the lactococcal vector pNZ18 could introduce and maintain four different genes for many generations in A. laidlawii. One of these, encoding the dominant membrane lipoprotein spiralin from the mollicute Spiroplasma citri, was expressed; however, expression was weak, the signal peptide of spiralin was not cleaved and the protein was not covalently modified by fatty acids. This resulted in a hydrophilic character of spiralin and its cytoplasmic localization in A. laidlawii. To increase the expression of foreign genes, random A. laidlawii DNA fragments were cloned into a pNZ18-related plasmid and expression signals were selected using the Bacillus licheniformis alpha amylase gene as a probe. Selection was done in Escherichia coli as well as directly in A. laidlawii. Active recombinants from E. coli were also able to express alpha-amylase activities and an enzyme of native size in A. laidlawii. The highest activity was obtained from a recombinant selected directly in A. laidlawii. This is the first example of a promoter sequence selected in a mollicute. Analysis of the putative promoters in seven clones revealed similar 10 and -35 regions, and similar spacer distances in A. laidlawii, Acholeplasma oculi, Lactococcus and E. coli. Vectors related to pNZ18 should be useful for the genetic analysis of specific A. laidlawii proteins and functions. PMID- 7496520 TI - The Cowdria ruminantium groE operon. AB - A Cowdria ruminantium genomic DNA library was constructed in the expression vector lambda ZAPII, and an immunoreactive clone, designated lambda Cr9.4, was isolated by screening with serum from a C. ruminantium-infected goat. Sequencing of the insert from this clone revealed two open reading frames, encoding peptides of 10462 and 58697 kDa respectively. Database searching indicated that the two genes were homologues of groES and groEL, genes encoding a group of heat shock proteins involved in protein processing, export and assembly. Western blotting experiments showed that the recombinant GroEL protein was recognized by sera raised against four isolates of C. ruminantium which originate from South Africa, West Africa and the Caribbean, but not by antisera to the closely related Ehrlichia species (E. ovina, E. [Cytoecetes] ondiri, E. bovis, E. phagocytophila) of African and European ruminants which can be expected to occur in similar geographical areas to C. ruminantium. This suggests that this protein may be useful in development of serodiagnostic tests for C. ruminantium infection which are not subject to cross-reactions with antibodies to Ehrlichia species. The cloning and expression of the GroE operon will also facilitate further study of the roles of the GroE proteins in the immune response to C. ruminantium. PMID- 7496519 TI - Delineation of the virulence-related locus (vrl) of Dichelobacter nodosus. AB - Dichelobacter nodosus is the primary pathogen implicated in ovine footrot. In this paper we have delineated a 27 kb locus, termed the virulence-related locus (vrl), that was essentially specific for virulent D. nodosus isolates. The precise ends of this locus were mapped and the sequences of the junction regions from the virulent strain A198 were compared to corresponding sequences from the benign isolate C305. The left end of the vrl locus was located in a sequence similar to that of the small stable 10Sa RNA molecule of Escherichia coli, next to a phage-attachment-site-like sequence, which indicated that the vrl locus might have arisen by the integration of a phage. However, no attachment-like sequence could be found at the right end of the vrl locus. In the chromosome of the benign strain the sequences bordering vrl were not contiguous but were separated by about 3 kb. It was concluded that the divergence of the benign and virulent strains at this locus was a multi-step process. Several potential ORFs were identified at the junction regions but only one ORF, encoding a 126 kDa protein, was expressed in a T7 expression system in E. coli. PMID- 7496521 TI - Induction of heat shock response in Vibrio cholerae. AB - General properties of the heat shock response in Vibrio cholerae were examined. Enhanced or de novo synthesis of 24 proteins was observed upon heat shock from 30 degrees C to 42 degrees C in cells labelled with [35S]methionine. A similar response could also be induced by a rise in temperature from 30 degrees C to 37 degrees C. Of these heat shock proteins, two were determined to be homologues of GroEL and DnaK, based upon their immunological cross-reactivity with antibodies raised against the Escherichia coli proteins. Three proteins, of molecular sizes 38, 44 and 48 kDa, which were undetectable in the 30 degrees C grown culture, appeared de novo after the heat shock. As in other prokaryotic systems thermal induction of many of the proteins was transient, but both DnaK and GroEL remained induced for at least 28 min after heat shock. DNA hybridization studies revealed that genes analogous not only to dnaK and groEL but also to dnaJ of E. coli exist in V. cholerae. Heat shock induced thermotolerance in V. cholerae but made the cells more sensitive to UV radiation. Unlike in E. coli, however, heat shock had no effect on the progeny virus yield in V. cholerae. PMID- 7496522 TI - Restriction site polymorphism of the genes encoding the major 25 kDa and 36 kDa outer-membrane proteins of Brucella. AB - Seventy-seven Brucella reference and field strains from different geographic origins and hosts representing the six recognized species and their different biovars were analysed for diversity of their genes encoding the major 25 and 36 kDa outer-membrane proteins (OMPs) by PCR-RFLP. The 25 kDa OMP is encoded by a single gene (omp25) whereas two closely related genes (omp2a and omp2b) encode and potentially express the 36 kDa OMP. Analysis of PCR products of the omp25 gene digested with nine restriction enzymes revealed two species-specific markers, i.e. the absence of the EcoRV site in all Brucella melitensis strains and an approximately 50 bp deletion at the 3' terminal end of the gene in all Brucella ovis strains. Analysis of PCR products of the omp2a and omp2b genes digested with 13 restriction enzymes indicated a greater diversity than the omp25 gene among the six Brucella species and within the Brucella abortus, Brucella suis, B. melitensis and B. ovis species. Greater polymorphism was also detected for the omp2b than for the omp2a gene, especially in B. ovis which seemed to carry two similar (but not identical) copies of omp2a instead of one copy each of omp2a and omp2b for the other Brucella species as was previously suggested by Ficht et al. (1990; Mol Microbiol 4, 1135-1142). Results of PCR-RFLP indicated that distinction can be made between Brucellia species and some of their biovars, except between B. canis and B. suis bv. 3 and 4, on the basis of the size and diversity of their major OMP genes, and that it could be of importance for diagnostic, epidemiological and evolutionary study purposes. PMID- 7496523 TI - Characterization of the Mycobacterium tuberculosis erp gene encoding a potential cell surface protein with repetitive structures. AB - Using the phoA gene fusion methodology adapted to mycobacteria, several Mycobacterium tuberculosis DNA fragments encoding exported proteins were recently identified. In this paper, the molecular cloning, genomic positioning, nucleotide sequence determination and transcriptional start site mapping of a new M. tuberculosis gene, identified by this methodology, are reported. This gene was called erp (for exported repetitive protein) and has a sequence similar to that of the Mycobacterium leprae 28 kDa antigen irg gene M. tuberculosis erp gene contains a putative iron box close to the mapped transcriptional start site. The predicted Erp protein displays a typical N-terminal signal sequence, a hydrophobic domain at the C-terminus and harbours repeated amino acid motifs. These structural features are reminiscent of cell-wall-associated surface proteins from Gram-positive bacteria. We found that these repeats are conserved among M. tuberculosis isolates, and are absent from the published M. leprae irg gene sequence. In addition to being present in M. leprae, erp sequences were found in other members of the M. tuberculosis complex, but not in other mycobacteria tested. These results suggest that erp might encode a cell surface component shared by major pathogenic mycobacteria. PMID- 7496524 TI - Species-specific identification of Mycobacterium bovis by PCR. AB - The Random Amplified Polymorphic DNA (RAPD) technique was used in the identification of a species-specific fragment of Mycobacterium bovis. A fragment of approximately 500 bp was amplified from the genome of 15 different M. bovis strains, including M. bovis BCG Pasteur, but was shown to be absent in 26 different mycobacteria and 20 different clinical isolates of Mycobacterium tuberculosis. When the fragment was used as a probe in a Southern blot analysis, several radioactive bands common to M. tuberculosis and M. bovis were observed. However, this fragment hybridized specifically to a 2900 bp EcoRI fragment in the M. bovis genome, but failed to hybridize in either M. tuberculosis or M. avium chromosomal DNA. Based on a partial nucleotide sequence of the 500 bp fragment, two oligonucleotide primers were designed and a PCR assay was developed. Using purified mycobacterial DNA samples, only M. bovis and M. bovis BCG rendered a unique amplification band. This PCR assay is able to detect down to 10 fg purified M. bovis DNA, which corresponds roughly to two bacilli. The assay is also useful for identifying the bacilli directly from uncultured biological samples, such as milk. PMID- 7496525 TI - Nucleotide sequences of streptomycete 16S ribosomal DNA: towards a specific identification system for streptomycetes using PCR. AB - To facilitate the differential identification of the genus Streptomyces, the 16S rRNA genes of 17 actinomycetes were sequenced and screened for the existence of streptomycete-specific signatures. The 16S rDNA of the Streptomyces strains and Amycolatopsis orientalis subsp. lurida exhibited 95-100% similarity, while that of the 16S rDNA of Actinoplanes utahensis showed only 88% similarity to the streptomycete 16S rDNAs. Potential genus-specific sequences were found in regions located around nucleotide positions 120, 800 and 1100. Several sets of primers derived from these characteristic regions were investigated as to their specificity in PCR-mediated amplifications. Most sets allowed selective amplification of the streptomycete rDNA sequences studied. RFLPs in the 16S rDNA permitted all strains to be distinguished. PMID- 7496526 TI - Selective recovery of Streptosporangium fragile from soil by indirect immunomagnetic capture. AB - A polyclonal antibody raised to Streptosporangium fragile spores reacted strongly and specifically with the immunizing strain and to a number of related species of Streptosporangium, as determined by dot immunoblotting. An indirect immunomagnetic capture method was developed for the recovery of the target organism from sterile and non-sterile soil, using sheep anti-rabbit M-280 Dynabeads. The effects of different soil blocking agents, antibody labelling concentrations and spore/Dynabead capture times on the recovery of S. fragile spores were investigated. Pre-blocking of antibody binding sites within the soil, with either 2% partially hydrolysed gelatin or 10% skimmed milk, was essential prior to immunomagnetic capture. Increasing the capture time from 15 to 60 min did not affect spore recovery; however, a 10-fold decline in the magnetic bead concentration did result in a significantly lower recovery of spores from soil. S. fragile was selectively enriched (1:190-fold) when present as a mixed population with Arthrobacter oxydans in sterile soil. The indirect immunomagnetic capture method was used to selectively recover S. fragile spores seeded into non sterile soil, although some background binding of non-target bacteria was noted. The target was successfully recovered from a sterile soil microcosm after 14 d incubation and the capture rate was increased by the inclusion of an initial soil dispersion and biomass concentration procedure, using the ion-exchange resin Chelex 100. PMID- 7496527 TI - Distribution of the ardA family of antirestriction genes on conjugative plasmids. AB - The ardA gene of I1 plasmid ColIb-P9 was previously shown to alleviate DNA restriction by type I enzymes and to promote conjugative transmission of the unmodified plasmid to a restricting host. To clarify the ecological role of ardA, its distribution was determined on plasmids from 23 incompatibility groups using hybridization to the coding sequence as an assay. Hybridizing sequences, shown by nucleotide sequencing to have at least 60% identity with ardA, were detected on plasmids belonging to the I complex (IncB, I1 and K), the F complex (IncFV) and the IncN group. The ardA homologues were found to specify an antirestriction phenotype which was enhanced by genetic depression of the plasmid transfer system. ardA loci map in plasmid leading regions but show no consistent association with a particular type of origin-of-transfer or a leading region gene of the ssb (single-stranded DNA-binding protein), psiB (plasmid SOS inhibition) and hok (host killing) families. It may be significant that ardA+ plasmids are authentic enterobacterial plasmids and that type I restriction systems are associated historically with members of the Enterobacteriaceae. PMID- 7496528 TI - IS1500, an IS3-like element from Leptospira interrogans. AB - Copies of an insertion-sequence (IS)-like element were isolated from two closely related serovars of Leptospira interrogans sensu stricto. Nucleotide sequence analysis of the 1236 bp element showed a characteristic IS structure with terminal imperfect inverted repeats (IRs) flanking a 1159 bp central region. This element was designated IS1500. Four open reading frames (orfA-orfD) were found in the central 'unique' region of IS1500. Similarities were detected between ORFA and ORFB and the putative transposases from members of the IS3 family of transposable elements. IS1500 or IS1500-like sequences were also detected in all other pathogenic leptospiral serovars, but not in the saprophytic species L. biflexa. Differences in IS1500 copy numbers in members of the same species suggest that this element can transpose. Physical mapping of IS1500 insertions in L. interrogans serovars icterohaemorrhagiae and pomona showed insertions were only on the large chromosomal replicon. The location of some IS1500 insertions coincides with regions of the genome that have undergone large rearrangements. PMID- 7496529 TI - Molecular genetic analysis of a thioredoxin gene from Thiobacillus ferrooxidans. AB - The Thiobacillus ferrooxidans thioredoxin gene, trxA, was isolated by its ability to complement an Escherichia coli gshA trxA mutant which was otherwise unable to grow on minimal medium lacking glutathione. The T. ferrooxidans thioredoxin also enabled the in vivo reduction by E. coli of methionine sulfoxide to methionine, as well as the in vitro reduction of insulin. When present in E. coli, the T. ferrooxidans thioredoxin supported the replication of phage T7, but not the growth of phage M13. The T. ferrooxidans trxA gene was sequenced and the thioredoxin was found to be most like that of E. coli (71% identity) and Chromatium vinosum (70% identity). As in the case of E. coli, the gene was located immediately upstream of the gene for the rho transcriptional terminator. DNA:RNA blot hybridization and primer-extension analysis of the trxA gene in T. ferrooxidans and the cloned gene in E. coli indicated that it was transcribed as an independent unit and that the major transcriptional start sites were the same in both organisms. PMID- 7496530 TI - A group I intron in the terminase gene of Lactobacillus delbrueckii subsp. lactis phage LL-H. AB - An 837 nt long group IA intron was discovered in the Lactobacillus delbrueckii subsp. lactis virulent phage LL-H genome. The LL-H intron conforms well to the secondary structure that is common to all group I introns. The only exception is that the extreme 3' nucleotide of the intron is an A residue instead of the usual G; despite this the intron is efficiently spliced in vivo. This LL-H intron contains an ORF, ORF168, which shows homology with endonucleases encoded by ORFs contained in Bacillus subtilis phage introns. At present, the LL-H intron is the only one found in the phages of lactic acid bacteria and the first one to be found in a phage belonging to the most abundant taxonomic group, group B or Siphoviridae. The LL-H intron interrupts gene terL, the product of which (50.5 kDa, TerL) is significantly homologous to the large subunit of B. subtilis phage SPP1 terminase. The product of the upstream gene, terS of LL-H (15.9 kDa, TerS), shows homology to small subunits of B. subtilis phage terminases. PMID- 7496531 TI - The bacteriophage 434 operator/repressor system in yeast. AB - The ability of the bacteriophage 434 operator/repressor system to function in a eukaryotic cell has been explored. An idealized 434 operator was placed at various positions in the PGK promoter of Saccharomyces cerevisiae: within the upstream activator sequence, close to the TATA box, and downstream of the transcription-initiation site. Expression of the 434 cI gene from a 2 microns based plasmid resulted in significant repression of gene expression from constructs containing the altered promoters linked to a beta-galactosidase reporter gene. Attempts to use a variant of the 434 repressor that has the binding specificity of the P22 repressor (434P22) were unsuccessful, due to a severely inhibitory effect of this gene-product on the growth of the yeast cells. PMID- 7496532 TI - IMP2, a gene involved in the expression of glucose-repressible genes in Saccharomyces cerevisiae. AB - Two mutants carrying different deletions of the IMP2 coding sequence of Saccharomyces cerevisiae, delta T1, which encodes a protein lacking the last 26 C terminal amino acids, and delta T2, which completely lacks the coding region, were analysed for derepression of glucose-repressible maltose, galactose, raffinose and ethanol utilization pathways in response to glucose limitation. The role of the IMP2 gene product in the regulation of carbon catabolite repressible enzymes maltase, invertase, alcohol dehydrogenase, NAD-dependent glutamate dehydrogenase (NAD-GDH) and L-lactate:ferricytochrome-c oxidoreductase (L-LCR) was also analysed. The IMP2 gene product is required for the rapid glucose derepression of all above-mentioned carbon source utilization pathways and of all the enzymes except for L-LCR. NAD-GDH is regulated by IMP2 in the opposite way and, in fact, this enzyme was released at higher levels in both imp2 mutants than in the wild-type strain. Therefore, the product of IMP2 appears to be involved in positive and negative regulation. Both deletions result in growth and catalytic defects; in some cases partial modification of the gene product yielded more dramatic effects than its complete absence. Moreover, evidence is provided that the IMP2 gene product regulates galactose- and maltose-inducible genes at the transcriptional level and is a positive regulator of maltase, maltose permease and galactose permease gene expression. PMID- 7496533 TI - Functional analysis of the Bacillus subtilis purT gene encoding formate-dependent glycinamide ribonucleotide transformylase. AB - The purT gene from Bacillus subtilis encoding the formate-dependent glycinamide ribonucleotide transformylase T was cloned by functional complementation of an Escherichia coli purN purT double mutant. The nucleotide sequence revealed an open reading frame of 384 amino acids. The purT amino acid sequence showed similarity to the enzyme phosphoribosylaminoimidazole carboxylase encoded by the purK gene but not to the N10-formyltetrahydrofolate-dependent glycinamide ribonucleotide transformylase N enzyme encoded by the purN gene. The glycinamide ribonucleotide transformylase T level was repressed in cells grown in rich medium compared to minimal-medium-grown cells. However, when the culture entered the stationary-growth phase the enzyme level increased in rich medium and decreased in minimal medium. By comparing the deduced amino acid sequence of the B. subtilis purT gene product with translated nucleotide sequences in various databanks, evidence for the existence of putative purT genes in the Gram-negative bacteria Pasteurella haemolytica and Pseudomonas aeruginosa was obtained. PMID- 7496534 TI - Identical amino acid sequence of the aroA(G) gene products of Bacillus subtilis 168 and B. subtilis Marburg strain. AB - A DNA fragment containing the aroA(G) gene of Bacillus subtilis 168, encoding 3 deoxy-D-arabino-heptulosonate 7-phosphate (DAHP) synthase-chorismate mutase, was cloned and sequenced. The N-terminus of the protein encoded by aroA(G) showed homology with chorismate mutase encoded by aroH of B. subtilis and with the chorismate mutase parts of proteins encoded by the pheA and tyrA genes of Escherichia coli. The C-terminus of the aroA(G) product has sequence similarity with 3-deoxy-D-manno-octulosonate 8-phosphate synthase of E. coli. It was shown that the proteins encoded by the aroA(G) gene of B. subtilis 168 and the aroA gene of B. subtilis ATCC 6051 Marburg strain are identical, so the observed differences in DAHP synthase activity from these two strains must result from other changes. PMID- 7496536 TI - Characterization of a nitrogen-fixation (nif) gene cluster from Anabaena azollae 1a shows that closely related cyanobacteria have highly variable but structured intergenic regions. AB - The exact identity of cyanobacteria that have been cultured from symbiotic associations with the water fern Azolla spp., whether they are required in the symbiotic process, and their relationship to the symbiotic species, is a matter of some debate. We have characterized a 6 kb region containing the nifB operon and the nifH gene from cyanobacterium Anabaena azollae 1a, a putative symbiont of Azolla caroliniana. Five complete open reading frames have been sequenced. All are very highly conserved when compared with the corresponding regions of Anabaena sp. PCC 7120, with 93% to 97% identity at the nucleotide level and 93% to almost 100% at the amino-acid level. The intergenic regions, however, are not highly conserved (53-89% identity) when compared to the corresponding regions of Anabaena 7120: the A. azollae genome contains both more copies and more types of short tandemly repeated repetitive sequences than Anabaena 7120. The start points of transcription for both the nifB and nifH operons were mapped and found to be the same as those in Anabaena 7120. It was not possible to discern an improved consensus nif promoter sequence, but it was possible to define the likely extent of the promoter to within 40 bases upstream of the transcription start-point. PMID- 7496535 TI - The 4-hydroxy-2-oxovalerate aldolase and acetaldehyde dehydrogenase (acylating) encoded by the nahM and nahO genes of the naphthalene catabolic plasmid pWW60-22 provide further evidence of conservation of meta-cleavage pathway gene sequences. AB - We report the complete nucleotide sequence and over-expression of the nahOM genes for the acetaldehyde dehydrogenase (acylating) and the 4-hydroxy-2-oxovalerate aldolase from the meta pathway operon of the naphthalene catabolic plasmid pWW60 22 from Pseudomonas sp. NCIMB9816. Additional partial sequence analysis of adjacent DNA shows the gene order within the operon to be nahNLOMK, identical to the order found for the isofunctional genes in the meta pathway operons in the toluene/xylene pathway of TOL plasmid pWW0 and the phenol/methylphenol pathway of pVI150. The deduced amino acid sequences of NahO and NahM were significantly homologous to the equivalent enzymes encoded by other Pseudomonas meta pathways, although both were the most divergent in each comparison. The nahOM genes were inserted downstream of the T7 promoter in the expression vector pET3a and similar constructs were also made of the isofunctional regions from pVI150 (dmpFG) and TOL plasmid pDK1 (xyIQK). High expression of all three gene pairs was detected by enzyme assays and by SDS-PAGE. PMID- 7496537 TI - Fructose phosphotransferase system of Xanthomonas campestris pv. campestris: characterization of the fruB gene. AB - In the plant pathogen Xanthomonas campestris pv. campestris, fructose is transported by a specific phosphotransferase system (PTS). This PTS involves a multiphosphoryl transfer protein (MTP) encoded by the fruB gene, which was cloned and sequenced. fruB is part of a transcriptional unit together with the fruK gene, coding for 1-phosphofructokinase, which is located upstream from the fruA gene, coding for the fructose-specific permease (EIIB'BCFru). The amino acid sequence of the X. campestris MTP deduced from the fruB sequence shared 46% identical residues with an MTP identified in Rhodobacter capsulatus. The X. campestris MTP (837 amino acid residues) consists of three moieties: a fructose specific enzyme-IIA-like N-terminal moiety (residues 1-148), followed by an HPr like moiety (161-251) and an enzyme-I-like C-terminal moiety (274-837). The three domains are separated by two flexible hinge regions rich in proline and alanine residues. The construction of a fruB mutant confirmed the role of the MTP in fructose transport and phosphorylation in X. campestris. PMID- 7496538 TI - Regulation of the lactose phosphotransferase system of Streptococcus bovis by glucose: independence of inducer exclusion and expulsion mechanisms. AB - Streptococcus bovis had a diauxic pattern of glucose and lactose utilization, and both of these sugars were transported by the sugar phosphotransferase system (PTS). Lactose catabolism was inducible, and S. bovis used the tagatose pathway to ferment lactose. Since a mutant that was deficient in glucose PTS activity transported lactose as fast as the wild-type, it appeared that S. bovis has separate enzyme IIs for glucose and lactose. The nonmetabolizable glucose analogue 2-deoxyglucose (2-DG) was a noncompetitive inhibitor of methyl beta-D thiogalactopyranoside (TMG) transport, and cells that were provided with either glucose or 2-DG were unable to transport TMG or lactose. Because the glucose-PTS deficient mutant could ferment glucose, but could not exclude TMG, it appeared that enzyme IIGlc rather than glucose catabolism per se was the critical feature of inducer exclusion. Cells that had accumulated TMG as TMG 6-phosphate expelled free TMG when glucose was added, but 2-DG was unable to cause TMG expulsion. The glucose-PTS-deficient mutant could still expel TMG in the presence of exogenous glucose. Membrane vesicles also exhibited glucose-dependent TMG exclusion and TMG expulsion. Membrane vesicles that were electroporated with phosphoenolpyruvate (PEP) and HPr retained TMG for more than 3 min, but vesicles that were electroporated with PEP plus HPr and fructose 1,6-diphosphate (FDP) (or glycerate 2-phosphate) lost their ability to retain TMG. Because FDP was able to trigger the ATP-dependent phosphorylation of HPr, it appeared that inducer expulsion was mediated by an FDP-activated protein kinase.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7496539 TI - Bacterial [Cu,Zn]-superoxide dismutase: phylogenetically distinct from the eukaryotic enzyme, and not so rare after all! AB - Copper- and zinc-containing superoxide dismutases ([Cu,Zn]-SODs) are generally considered almost exclusively eukaryotic enzymes, protecting the cytosol and extracellular compartments of higher organisms from damage by oxygen free radicals. The recent description of a few examples of bacterial forms of the enzyme, located in the periplasm of different Gram-negative micro-organisms, prompted a re-evaluation of this general perception. A PCR-based approach has been developed and used successfully to identify bacterial genes encoding [Cu,Zn] SOD in a wide range of important human and animal pathogens-members of the Haemophilus, Actinobacillus and Pasteurella (HAP) group, and Neisseria meningitidis. Comparison of [Cu,Zn]-SOD peptide sequences found in Haemophilus ducreyi, Actinobacillus pleuropneumoniae, Actinobacillus actinomycetemcomitans, Pasteurella multocida, and N. meningitidis with previously described bacterial proteins and examples of eukaryotic [Cu,Zn]-SOD has shown that the bacterial proteins constitute a distinct family apparently widely separated in evolutionary terms from the eukaryotic examples. The widespread occurrence of [Cu,Zn]-SOD in the periplasm of bacterial pathogens, appropriately located to dismute exogenously derived superoxide radical anions, suggests that this enzyme may play a role in the interactive biology of organisms with their hosts and so contribute to their capacity to cause disease. PMID- 7496540 TI - Biosynthesis of glycoproteins in Candida albicans: activity of dolichol phosphate mannose synthase and protein mannosylation in a mixed membrane fraction. AB - A mixed membrane fraction (MMF) was isolated from yeast cells of Candida albicans with the ability to synthesize dolichol phosphate mannose (Dol-P-Man) from GDP Man and dolichol phosphate (Dol-P) and transfer the sugar to proteins. Temperature of incubation (20-37 degrees C) did not affect the synthesis of Dol-P Man but protein mannosylation occurred better at physiological temperatures (28 degrees C and 37 degrees C). Most of the sugar (87-93%) in the mannoproteins was O-linked as judged by its release by beta-elimination. Mannose was identified as the sole product after this treatment. Following incubation of MMF with the sugar donor, parallel levels of Dol-P-Man and mannosylated proteins were detected up to 30 min. Thereafter, Dol-P-Man levels reached a steady value whereas mannoproteins rapidly accumulated. Lipid-linked oligosaccharides were also detected in incubation mixtures, though in much lower amounts than those of Dol-P-Man or mannoproteins. Dol-P-Man synthase activity increased proportionally in response to increasing concentrations of either of the two enzyme substrates. A Km value of 0.36 microM for GDP-Man was calculated. MMF failed to use exogenous Dol-P-Man for protein glycosylation. Specific inhibition of Dol-P-Man synthesis with amphomycin was concomitant with a parallel decrease in protein mannosylation, indicating that most of the sugar is transferred to protein via the carrier lipid. Results are discussed in terms of the role of Dol-P-Man in protein glycosylation in C. albicans. PMID- 7496541 TI - Substrate specificity of nine NAD(+)-dependent alcohol dehydrogenases in Aspergillus nidulans. AB - In Aspergillus nidulans three alcohol dehydrogenases (ADHs) have been described. ADHI is induced by ethanol and is the physiological enzyme of ethanol utilization, ADHII has not been attributed a function but is repressed by ethanol. The ALCR regulatory protein acts positively to induce ADHI, and negatively in its control of ADHII. ADHIII is specifically induced by anaerobic stress. We have characterized the substrate specificity of these three enzymes by looking at their staining profile on polyacrylamide gels with a range of alcohols. In addition to these enzymes we have observed six other NAD(+) dependent ADHs, two of which, propan-2-ol dehydrogenase and pentan-2-ol dehydrogenase, share similar control with ADHII. The inducibility of these enzymes with some alcohols has also been investigated. The profile of ADHs with NADP+ as an electron acceptor is also reported. PMID- 7496542 TI - An extreme creA mutation in Aspergillus nidulans has severe effects on D-glucose utilization. AB - Aspergillus nidulans wild-type and the extreme carbon catabolite derepressed mutant creAd-30 were characterized with respect to enzyme activities, metabolite concentrations and polyol pools all related to glycolysis, after growth on D glucose. In the creAd-30 strain the enzymes hexokinase and fructose-6-phosphate reductase showed a two- and threefold increase in activity, respectively, whereas phosphofructokinase and pyruvate kinase activity decreased two- and threefold, respectively, in comparison with the wild-type strain. The most notable changes in metabolite concentrations were that fructose 2,6-bisphosphate and fructose 1,6 bisphosphate showed a 2.5-fold increase, whereas both pyruvate and citrate decreased in the creAd-30. Striking differences were found for the polyol concentrations measured for the two strains tested. Intracellular glycerol and arabitol concentrations were 10-fold higher and erythritol fivefold higher in creAd-30, whereas intracellular trehalose and mannitol were both decreased. The total internal polyol concentration appears to be constant at approximately 700 mumol (g dry wt)-1. All polyols were also detected in high amounts in the culture filtrate of the creAd-30 mutant strain but no extracellular trehalose was found. The overall production of polyols in this strain was therefore much higher than in the wild-type. The high level of polyols produced and the changes in metabolite concentrations in the creAd-30 strain suggest that the differences in enzyme activities result in an altered flow through glycolysis leading to a more rapid formation of polyols which are subsequently secreted. PMID- 7496544 TI - Glucose metabolism in 'Sphingomonas elodea': pathway engineering via construction of a glucose-6-phosphate dehydrogenase insertion mutant. AB - 'Sphingomonas (formerly Pseudomonas) elodea' produces the industrially important polysaccharide gellan when grown in media containing glucose. Glucose catabolic enzymes and enzymes of central carbon metabolism were assayed in crude extracts of glucose-grown cultures of this bacterium. Based on these analyses it was concluded that glucose is converted to either gluconate or glucose 6-phosphate and that both of these products are converted to 6-phosphogluconate, a precursor for the Entner-Doudoroff (ED) and pentose phosphate pathways. Phosphoglucoisomerase (Pgi) activity was detected, but the lack of phosphofructokinase activity indicated that the Embden-Meyerhof glycolytic pathway is non-functional for glucose degradation. Thus, this bacterium utilizes glucose mainly via the ED and pentose phosphate pathways. Enzyme analyses suggested the involvement of glucose-6-phosphate dehydrogenase (Zwf) in glucose utilization and CO2 production. The zwf gene was cloned from 'S. elodea' and partially sequenced, and a null zwf mutant was constructed. This mutant exhibited no Zwf activity in in vitro assays, grew normally on glucose minimal medium and accumulated biomass (cells plus gellan) and produced CO2 at the same rates as the parental strain. Potential explanations for this finding are provided. Clones carrying the pgi gene were isolated fortuitously. PMID- 7496543 TI - Physarum polycephalum haemagglutinins: effect of nutrition on synthesis, and their possible role in nature. AB - The activity of haemagglutinins in plasmodia of Physarum polycephalum was measured under different culture conditions. The activity was markedly increased when the plasmodia were incubated in a non-nutrient salt medium. During starvation, significant amounts of haemagglutinins were found in the slime layer on the surface of the plasmodia. An increase in activity was not observed in the presence of actinomycin D or cycloheximide. Under starvation conditions, plasmodia are known to differentiate into either sclerotia (spherules) or fruiting bodies. Acceleration of haemagglutinin synthesis, however, was not always observed during spherulation and fruiting-body formation. Attempts to detect endogenous glycoconjugates that bind to the haemagglutinins were unsuccessful but we found that the haemagglutinins could bind to acidic polysaccharides produced by Escherichia coli K12. The bacterial glycoconjugates were purified and partially characterized. They contained N-acetylhexosamine residues which appeared to be important for binding with the haemagglutinins. It is possible that the haemagglutinins play a physiological role in the interaction with these organisms. PMID- 7496545 TI - Trauma, borderline personality, and self-harm behaviors. PMID- 7496547 TI - Walking the line. PMID- 7496546 TI - Diagnosis of microcytic anemia by urinalysis. PMID- 7496548 TI - The elements of statistics for clinical readers. AB - Statistical courses usually emphasize teaching the mathematical properties of methods to perform statistical significance testing. To interpret clinical studies, however, the reader needs to know only the purpose of the statistical methods, not their mathematical basis. This purpose in most studies is to evaluate the association between a risk factor and an outcome. The evaluation has five components: (1) measure the strength of the association, (2) determine the probability that the observed association did not occur by chance alone, (3) find the range of probable values for the measure of association, (4) reduce the possibility that the association is invalid because of confounding factors, and (5) examine the possibility that the association does not apply equally well to all people because of modulating factors. By focusing on the purposes of the statistical evaluation, the reader will be less distracted by the specific mathematical formulations that provide little additional information to the clinician. PMID- 7496549 TI - New knowledge, old patients. PMID- 7496550 TI - Use of warfarin for nonvalvular atrial fibrillation in nursing home patients. AB - OBJECTIVE: To assess warfarin sodium use and anticoagulant monitoring in nursing home patients with nonvalvular atrial fibrillation (NVAF), according to American College of Chest Physicians Consensus Conference guidelines. DESIGN: Retrospective, multicenter, point-prevalence study. SETTING: Nursing homes in Minneapolis-St Paul, Minn. PATIENTS: Nine-hundred two patients 60 years and older, from whom 69 with a diagnosis of NVAF and 16 with valvular atrial fibrillation (VAF) (control group) were identified. DATA COLLECTED: Patient demographics and diseases, diagnostic tests for atrial fibrillation (AF), antithrombotic drugs and dosage, anticoagulant activity test results, other drugs, and drug allergies were determined by chart review and attending physician response to written communication from the nursing home's medical director and consultant pharmacist. MAIN OUTCOME MEASURES: Prevalence of NVAF, VAF, and risk factors for thromboembolism and major bleeding, use of warfarin and other antithrombotic drugs for AF, anticoagulation control, and the relationship of warfarin dose with the recommended international normalized ratio (INR) and prothrombin time (PT). RESULTS: Nonvalvular AF was documented in 7.6% of the patients. Most patients with NVAF were at an increased risk for stroke, yet only 20% without a conventional contraindication to warfarin use experienced anticoagulation; a greater proportion of patients with VAF experienced anticoagulation. The INR was within the recommended range for NVAF over a 6-month period 37% of the time and recommended PT, 52% of the time. An equal percentage of warfarin dose changes occurred in response to a PT ratio outside the recommended range as occurred with an INR outside the recommended range. CONCLUSIONS: Many nursing home patients have NVAF with comorbid conditions that subject them to a greater than average risk for thromboembolic stroke. Warfarin is underused for stroke prophylaxis and often is not used according to the American College of Chest Physician guidelines. Physicians, nurses, and consultant pharmacists must be better informed about (1) known risk factors for thromboembolism and major bleeding to identify patients with AF who will most likely benefit from warfarin therapy, (2) maintaining an INR of 2 to 3, and (3) the need for small warfarin dose adjustments in elderly patients. PMID- 7496552 TI - Pulsed dye laser treatment of warts. AB - OBJECTIVE: To determine the safety and efficacy of the flashlamp-pumped pulsed dye laser for the treatment of uncomplicated and recalcitrant warts. DESIGN: Observational study. PATIENTS: A total of 142 patients with 703 recalcitrant and 25 previously untreated warts consecutively referred to two tertiary care laser centers from July 1993 through January 1994. INTERVENTION: Photocoagulation with the flashlamp-pumped pulsed dye laser. MAIN OUTCOME MEASURES: Complete resolution of treated warts after a follow-up ranging from 3 to 9 months. RESULTS: The overall response rates were 99% for body, limb, and anogenital warts; 95% for hand warts; 84% for plantar warts; and 83% for periungual warts. Side effects were limited and infrequent. CONCLUSION: Pulsed dye laser therapy is a highly effective and safe method used to selectively destroy warts without damaging the surrounding skin. PMID- 7496551 TI - Interruption in the medical interaction. AB - OBJECTIVES: To describe patterns of interruption in the physician-patient interaction; specifically, to determine who interrupts, to determine if a shift in control occurs as a result of interruption, and to characterize the information gained when patients interrupt physicians. DESIGN: Observational. SETTING: Community-based primary care practices in North Carolina. SUBJECTS: Internists and family physicians in private practice in North Carolina (six men and two women) and their patients (13 men and 27 women). INTERVENTIONS: None. MAIN OUTCOME MEASURE: The obtainment of control of the interaction, at least momentarily, as a result of interruption. RESULTS: Analysis of 40 audiotaped interactions revealed 833 interruptions (mean +/- SD, 20.8 +/- 12.2 per interaction): Patients initiated 55% of all interruptions. Physicians and patients each gained control of the conversation after 50% of interruptions. Patients gained control after 74% of patient-initiated interruptions, and physicians gained control after 79% of physician-initiated interruptions. Patients were more likely to gain control by interrupting late in the interaction, and 75% of patient-initiated interruptions resulted in new information (solicited and unsolicited) being contributed to the interaction. CONCLUSION: Interruption by patients can be an informative event. PMID- 7496553 TI - Human immunodeficiency virus seroprevalence in community-based primary care practices, 1990-1992. A report from the Ambulatory Sentinel Practice Network. AB - OBJECTIVE: To estimate the seroprevalence of human immunodeficiency virus type 1 (HIV-1) infection in primary care practices. METHODS: Fifty-four practices in the United States participated in an anonymous, unlinked HIV seroprevalence study between January 1990 and December 1992. Residual blood samples drawn for routine clinical tests from patients 15 to 49 years of age were centrally tested for the HIV-1 antibody for 1 month of each quarter. Information about patient demographics, clinician-recognized risk factors, the known HIV status of the patient, and whether the blood was drawn for HIV testing was recorded with each specimen. RESULTS: Of 21,998 specimens collected, 99 (0.45%) were seropositive. Of these 99 seropositive persons, 31.3% (a seroprevalence of 0.15%) were not suspected by their clinicians of being infected with HIV. Seroprevalences in men (0.96%) exceeded those in women (0.22%), and rates in rural practices (0.18%) were lower than in urban practices (0.71%). Among patients with unsuspected HIV infection, however, the gender differences, especially in rural areas, were less pronounced. Risk factors for HIV infection were infrequently noted. There was an increase in the overall seroprevalence during the 1990 to 1992 study period (0.36% to 0.53%); however, this trend was not statistically significant. CONCLUSIONS: Within a 3-year period, clinicians in at least two of five primary care practices can expect to encounter patients infected with HIV, regardless of practice location. Also, nearly one third of the patients with HIV infection will not be suspected of having this condition by their clinician. PMID- 7496554 TI - Practical management of osteoarthritis. Integration of pharmacologic and nonpharmacologic measures. AB - Pharmacotherapy should play an adjunctive role to nonpharmacologic measures in the overall management of patients with symptoms due to osteoarthritis. Patients should be instructed in how to rest or unload involved joints; protect them through appropriate manipulation of their environment and appropriate methods of lifting and bending; and maintain and improve muscle strength and flexibility to ensure joint stability and prevent contractures. In most patients, acetaminophen in doses of up to 4 g/d is preferable to nonsteroidal anti-inflammatory drugs (NSAIDs) as a first-choice agent because of its lower toxicity. If NSAIDs are used, they should be prescribed initially in lower (analgesic) doses. Ibuprofen may be the preferred initial NSAID because it can be given in low doses for short durations, has been associated with lower rates of gastrointestinal tract side effects, and is inexpensive. PMID- 7496556 TI - Maintaining communication in a time of uncertainty. AB - Today, the move to managed care and ever-ballooning technology have created new challenges for communication between physician and patient. Good communication involves a set of acquired skills that medical schools and continuing medical education programs are finally beginning to teach. One model for mastering these skills is known as the four Es: engage, empathize, educate, and enlist. The four E model, created by the Bayer (formerly Miles) Institute for Health Care Communication in West Haven, Conn, includes techniques that can help improve a physician's ability to communicate with patients. PMID- 7496555 TI - Nonsurgical management of osteoarthritis, with an emphasis on nonpharmacologic measures. AB - Osteoarthritis is treated primarily with nonsteroidal anti-inflammatory drugs. Concern is growing, however, among physicians and patients about the gastrointestinal side effects of these agents. Nonpharmacologic measures are underused in the treatment of osteoarthritis; isometric exercise and provision of social support may be as effective as nonsteroidal anti-inflammatory drugs in reducing pain and improving mobility in patients with knee osteoarthritis. Recent data argue persuasively for a change in our strategy for treatment of patients with osteoarthritis: the treatment program should emphasize nonpharmacologic measures, with drugs used in an adjunctive role. Only when this approach is not effective should nonsteroidal anti-inflammatory drugs be prescribed, and then, in the lowest dose needed for symptomatic relief. PMID- 7496557 TI - Nocturnal hypertension--under the cover of darkness. PMID- 7496558 TI - The role of platelets in essential hypertension. AB - Recent research is helping us understand the complex interactions that occur between platelets and their environment. The several intracellular events that occur during platelet activation are being identified as ar their effects on other platelets, the endothelium and coagulation factors. Heightened platelet activation is seen early in essential hypertension and probably plays an important role in the initiation and progression of atherosclerosis and the disorders associated with it. This review identifies some of the changes in platelet structure and function in essential hypertension and their role in the pathogenesis of hypertensive vascular disease. PMID- 7496559 TI - Malignant phaeochromocytoma with high circulating DOPA, and clonidine suppressible noradrenaline. AB - Phaeochromocytoma, "perhaps the most fascinating of all tumors" [1], can present with a broad range of clinical manifestations [2]. Once suspected, the biochemical diagnosis is straightforward in most patients since plasma and urinary levels of noradrenaline and/or adrenaline, and urinary metabolites are well above those encountered in healthy subjects or patients with essential hypertension [3,4]. Exceptions to this general rule are well known, however, hence suppression tests have found favour, particularly in cases where catecholamine levels are within, or close to, the normal range [5-7]. We present a unique patient with malignant phaeochromocytoma whose plasma noradrenaline levels were in the high-normal range, and suppressed normally with clonidine administration. He had extremely high circulating levels of dihydroxyphenylalanine (dopa) which were not affected by clonidine, and different patterns of catecholamine concentrations in tumour tissue and plasma. PMID- 7496560 TI - Determinants of the awakening rise in systemic blood pressure in obstructive sleep apnea syndrome. AB - To investigate the factors responsible for the morning rise in blood pressure (BP) in obstructive sleep apnea syndrome (OSAS) we examined a group of 253 consecutive snorers or OSAS patients. On the basis of their AHI the patients were classified in four groups. BP was measured on the evening before sleep onset and on the following morning after 15 min of rest by a finger arterial pressure device (Finapres). In 150 subjects BP was monitored during the night by a Finapres device. In the morning BP increased in the patient group with an average difference of 9.9 +/- 0.5 mmHg for systolic (SBP) and 9.9 +/- 0.4 mmHg for diastolic pressure (DBP). The increase was significant in snorers and OSAS patients without differences between groups. The morning rise in SBP was related to diurnal values of SBP, age and AHI whereas the time spent in apnea and the diurnal values of DBP significantly contributed to the DBP increase. In the subgroup of 150 patients in whom BP was analyzed during sleep, the awakening increase was related to the absolute BP value during sleep and to the BP changes from wakefulness to sleep. The magnitude of the BP changes from evening to morning was not dependent on the degree of BP variability during sleep. We conclude that the awakening increase in BP in patients with snoring or OSAS may be mediated by the setting of pressure response to apnea or to mechanical effort during sleep. Anthropometric variables and diurnal cardiovascular setting may play an additional role in modulating the final pressure response to upper airway obstruction. PMID- 7496561 TI - Effect of autonomic blockade on heart rate and blood pressure in sleep apnea syndrome. AB - To determine the impact of the autonomic nervous system on hemodynamic responses to sleep apnea we studied 7 male patients (48 +/- 6 years, BMI = 29.7 +/- 2.9 kg/m2) with an apnea index = 33 + 11/h. In two hypertensives, therapy was withdrawn 2 weeks before the study. During a full night polysomnography EEG, EOG, EMG, respiration, oximetry, heart rate (HR) and blood pressure (BP) (Finapres 23OOE) were recorded. Placebo and 3 short-acting autonomic blockers were randomly i.v. administered: antimuscarinic (atropine 1 mg, A), beta 1 blocker (esmolol 40 mg, E), alpha blocker (nicergoline 2.5 mg, N). A total of 367 obstructive apneas in NREM sleep were studied. With atropine, HR was increased at the beginning of apnea (P1)(74.5 +/- 2 bpm vs 65.4 +/- 2 bpm with placebo) and at the end apnea (P3) (76.6 +/- 1.9 bpm vs 67.1 +/- 1.76 bpm). At the resumption of breathing (RB), HR still increased (83.5 +/- 1.5 bpm vs 80.1 +/- bpm). Nicergoline had no significant effect on HR but significantly decreased SBP (117.8 +/- 1.4 mmHg vs 127 +/- 1.6 with placebo, p < 0.001). Nevertheless, the rise in BP from P3 to RB remained significant (p < 0.001). Esmolol significantly decreased HR at P1 (68.1 +/- 1.9 bpm vs 73.9 +/- 1.9 bpm) and at RB (75.9 +/- 1.9 vs 81.6 +/- 1.8). With A, the rise in HR at end-apnea (P3) suggests a vagal hyperactivity during this phase which is blunted at resumption when HR is unchanged.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7496562 TI - Sleep structure in essential hypertensive patients: differences between dippers and non-dippers. AB - The objective of this study was to determine whether the macrostructure and microstructure of sleep were altered in non-dipper essential hypertensive patients. Patients included 9 non-dipper essential hypertensive patients and 10 dippers. We measured blood pressure beat-to-beat by Finapres and all stages of sleep by polysomnografically recording simultaneously during spontaneous nocturnal sleep. We analysed blood pressure pattern for 4-min long random periods while the patients were awake and during all stages of sleep; sleep-efficiency (SE), sleep-latency (SL), delta sleep-latency (delta-SL), REM sleep-latency (REM SL), St. 1, St.2, St.3, St.4 and REM duration and percentage (%) values, and microstructural aspects of sleep (arousal and microarousal temporisation and features). Dipper patients showed a fall in blood pressure (BP) greater than 10% in all stages of NREM sleep; in the non-dipper patients BP fell by less than 10% of waking values in all NREM stages. REM sleep as well as HR were similar in both groups during all stages of sleep. Non-dippers showed the same number of arousals but more microarousals than dippers (p < 0.001). During and after microarousals BP and HR increased in non-dippers, but showed light variation in dippers. Microarousals induced several stage shifts towards lighter sleep. For this reason non-dippers spent less time in stage 4 than dippers (p < 0.001). In conclusion, non-dipper essential hypertensive patients are a subset of patients with central sympathetic hyperactivity responsible for quantitative and qualitative alteration of sleep. PMID- 7496563 TI - Effect of enalapril on erythrocytosis in hypertensive patients with renal disease. AB - Treatment of hypertension with an angiotensin converting enzyme inhibitor (ACEI) may be associated with a decrease in haemoglobin concentration especially in patients with renal insufficiency. This open study in 19 patients with a variety of renal diseases with complicating hypertension investigated the effects of the ACEI, enalapril, on haemoglobin and plasma erythropoietin (EPO) concentrations. Blood samples were obtained at baseline and 2, 60 and 120 days after starting treatment with enalapril. By day 60 there was a significant decrease in mean haemoglobin concentration (mean decrease 7.4 g/l) that was sustained until day 120. Apart from a small, but significant, reduction by day 2, mean plasma EPO concentration remained constant throughout the study. The magnitude of the decrease in haemoglobin concentration was, however, significantly correlated with the baseline plasma creatinine concentration and creatinine clearance. These results suggested that the degree of renal insufficiency was important in determining the haematological response to ACE inhibition. While the mechanism of these changes remains unclear, our findings suggest that inhibition of the renin angiotensin system, rather than decreasing EPO production, may reduce the erythropoietic activity of the hormone. PMID- 7496564 TI - Normalization of structural cardiovascular changes during antihypertensive treatment with a regimen based on the ACE-inhibitor perindopril. AB - Untreated essential hypertension is associated with left ventricular hypertrophy (LVH) and structural changes in resistance vessels. The aim of this study was to establish the effect of perindopril based antihypertensive therapy on media thickness to lumen diameter (media:lumen) ratio of peripheral resistance vessels and left ventricular mass in essential hypertension. Twenty-five patients with newly diagnosed or poorly regulated essential hypertension were treated with perindopril. Insufficient treatment response (DBP > 90 mmHg) led to addition of isradipine, and hydralazine was used as a tertiary drug if necessary. Gluteal subcutaneous biopsies were taken surgically at baseline and after 9 months of successful treatment. Two small resistance arteries were isolated and mounted in a small vessel myograph, and media:lumen ratio (%) was measured under standardized conditions. Left ventricular mass was determined by echocardiography. Mean (SD) media:lumen ratio decreased from 9.8 (2.6) % to 7.8 (1.9) % (p < 0.05), while left ventricular mass decreased from 299 (75) g to 199 (53) g (p < 0.001). Correlation was found between changes in left ventricular mass index and media:lumen ratio (r = 0.62, p < 0.01). It is concluded that a perindopril based regimen efficiently normalizes resistance artery structure and left ventricular hypertrophy in essential hypertension within one year of treatment. The impact of these findings on the excess cardiovascular morbidity and mortality in arterial hypertension remains to be investigated. PMID- 7496565 TI - Hypertension is not related to suppressed lymphocyte proliferation but to elevated NO synthesis in vascular smooth muscle cells of borderline hypertensive rat. AB - Hypertensive individuals often exhibit immune abnormalities. We have previously reported that spontaneously hypertensive rats (SHR) had a severely depressed lymphocyte proliferation response caused by excessive nitric oxide (NO) from macrophages and vascular smooth muscle cells (VSMC). However, the development of hypertension was not correlated with the lymphocyte depression and elevated NO synthesis in macrophages. In this study, we investigated the effect of hypertension on lymphocytes and the NO synthesis system in borderline hypertensive rats (BHR). BHR became significantly hypertensive after receiving a high sodium diet. The proliferation response of lymphocytes in hypertensive BHR was similar to that of normotensive BHR fed a normal diet or of Wistar Kyoto rats (WKY). NO production in macrophages of hypertensive BHR was not different from that of normotensive BHR or WKY. However, NO production in VSMC was significantly elevated in hypertensive BHR. A positive correlation between blood pressure and VSMC NO production exists in hypertensive BHR. These results suggested that high blood pressure neither affected the lymphocyte function nor influenced the activation of NO synthesis in macrophages. Hypertension, however, may influence the activation of VSMC NO synthesis. In conclusion, hypertension is not causally associated with immune dysfunction as seen in SHR but is related to enhanced NO synthesis in VSMC. PMID- 7496566 TI - [Ocular complications in diabetes mellitus in Zaire]. AB - OBJECTIVE: to determine the incidence of ocular complications of diabetes in Zaire. METHODS: the study was based on an analysis of data from 382 diabetic patients seen from 1978 through 1989, and from Jan. 1990 through April 1990. Ocular complications were determined by routine ophthalmic examination including an inspection of adnexa, biomicroscopy, ophthalmoscopy. RESULTS: the average age of patients was 46 years the male to female ratio being 2:1. Ocular complications were found in 200 (52%) of 382 patients and included diabetic retinopathy (32%), cataract (18%) and other manifestations (vitreous hemorrhage, diabetic papillopathy, neovascular glaucoma, rubeosis iridis, keratitis, hordeolum) were rare (2%). Blindness caused by diabetes mellitus was found in 4% of patients and was due to diabetic retinopathy and cataract. CONCLUSION: Ocular complications seen in this study were comparable to those reported in Africa and the World. PMID- 7496567 TI - [Aids and chorioretinal opportunistic infections]. AB - Retrospective study of 19 cases of opportunistic infections of the chorioretina in patients with the Acquired Immunodeficiency Syndrome. We observed 14 cases of CMV retinitis, 2 cases of toxoplasmic chorioretinitis, 1 case of cryptococcal choroiditis and 2 cases of herpes zoster retinitis. Review of the clinical, angiographical and histopathological aspects of these infections. Review of the vital and visual prognosis after treatment. PMID- 7496568 TI - Central retinal artery occlusion after back surgery: a case report. AB - Central retinal artery occlusion can be caused by excessive extrinsic pressure on the eyeball during surgery. It is a well-known and severe complication after elective back surgery. The authors report a case of central retinal artery occlusion in an adult after posterior lumbar fusion in which a rectangular headrest was used. PMID- 7496569 TI - Surgical treatment of myogenic blepharoptosis. AB - We describe the surgical approach of two patients with myogenic blepharoptosis. The ptosis is caused by a glycogenosis type II (Pompe disease) in the first case and is due to a juvenile, chronic progressive external ophthalmoplegia (Kearns Sayre syndrome) in the second case. The first patient presented a unilateral ptosis. The ptosis in the second patient was bilateral and manifest. The eyelids could only be opened manually. A resection of the palpebral levator muscle was carried out under local anesthesia. The choice of the technique will be explained. The amount of levator resection was calculated considering the residual eyelid motility and Bell's phenomenon. Electronmicroscopy of the resected levator muscle will be discussed. PMID- 7496570 TI - [Small pupil and phacoemulsification]. AB - Miotics pupils are compatible with phacoemulsification. Several techniques have been proposed to enlarge the pupil. We present a multiple sphincterotomies technique which enlarges the pupil to a size compatible with phacoemulsification, and retains postoperatively the circular shape of the pupil as well as its functional role. PMID- 7496571 TI - Reliability of the Nidek NT-1000 non contact tonometer. AB - In this prospective study, we compared the intraocular pressure (IOP) readings of 100 patients measured with the Goldmann applanation tonometer and the Nidek NT 1000 pneumotonometer. The correlation coefficient between the Goldmann and Nidek readings was 0.86. On the average the pneumotonometer overestimated the intraocular pressure with 0.43 mm Hg. The Nidek NT-1000 non contact tonometer can be used for screening purposes provided an appropriately low IOP value is used to indicate the need for further assessment with the Goldman applanation tonometer. PMID- 7496572 TI - Clinical experience with the single-plate Molteno drainage implant. AB - The results of our first 18 Molteno implants placed for treatment of intractable glaucoma are presented. The mean follow-up was 10 months. Indications, surgical technique, postoperative course and complications are discussed. PMID- 7496573 TI - [Combined operations: trabeculectomy, crystalline lens extraction and insertion of a posterior chamber implant: a 3-year retrospective study]. AB - Thirty-four eyes of 29 patients with coexisting cataract and glaucoma that underwent glaucoma triple procedure between 1991 et 1994 were retrospectively analyzed. Mean follow-up was 9 months. A phacotrabeculectomy (PHACO-TRAB) was performed in 21 eyes, 13 eyes received extracapsular cataract extraction with trabculectomy (ECCE-TRAB). In 26 eyes, adjunctive 5-FluoroUracil subconjunctival injections were administrated. Our results showed a mean decrease from preoperative intraocular pressure (IOP) baseline of 10.5 mmHg at 3 months. (Mean preoperative IOP = 25.6 mmHg, mean IOP 3 months = 15.1 mmHg). The mean IOP decrease appears equivalent in the PHACOTRAB group compared to the ECCE-TRAB group, with a stable IOP result during the follow-up. Overall mean visual acuity improved from 0.16 preoperatively to 0.6 at 3 months with a slightly better result in the PHACO-TRAB group. Postoperative hyphema and fibrinous anterior uveitis were seen more often in the ECCE-TRAB than in the PHACO-TRAB group. This observation, along with the rapidity of the surgery, the quicker visual rehabilitation and the technique that is less disruptive to the conjunctiva are arguments in favour of phacotrabeculectomy whenever a combined procedure is indicated. PMID- 7496574 TI - [External sclerostomy using the THC:YAG Holmium laser in the treatment of glaucoma]. AB - Ab externo THC:YAG Holmium laser sclerostomy is an interesting alternative to standard full thickness filtering surgery. We report our retrospective experience over a total number of 38 sclerostomies between march 1993 and september 1994 on 32 patients (33 eyes) with primary or secondary glaucomas composed of 17 phakic and 16 aphakic and/or pseudophakic eyes. The mean patient age is 62.9 years (range: 10 to 90 years). The mean follow-up time is 7.5 months (1 to 18 months). We used a long pulsed (300 microsec) laser emitting in the near infrared (2.1 microns). A conjunctival stab incision was made 12 mm away from the sclerostomy site to allow entry of a 22 G probe which delivers energy at a right angle to the long axis of the fiber. We used 100 mJ per pulse laser energy delivered at a rate of 5 impacts per second. The mean delivered energy to produce a functional sclerostomy was 16.1 J. All patients received subconjunctival injections of 5 Fluorouracil (mean number of injections: 7.4). Criteria for success included an IOP of 22 mmH or less with or without additional medical treatment and a decrease of at least 30% of IOP when the preoperative IOP was 22 mmHg or less. The mean preoperative IOP was 35.8 mmHg (21 to 54 mmHg). By 3 months, the mean IOP of the 14 eyes over 19 successfully controlled was 15.2 mmHg (8 to 22 mmHg) with additional medical treatment in 3 cases. By 6 months, 11 eyes over 11 successfully controlled was 14.1 mmHg (6 to 21 mmHg) with additional medical treatment in 3 cases.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7496576 TI - [Surgical treatment of an idiopathic macular hole: preliminary results]. AB - Our preliminary results include 8 patients (8 eyes) presenting with an idiopathic stage III full-thickness macular hole (Gass classification), 6-month-old or less by history. Five eyes were examined after complete disappearance of the gas C3F8 injected in the vitreous cavity at completion of surgery. Anatomical success, defined as a macular hole with a 360 degree flat edge, or as a macular hole which is no more visible, was obtained in 4 cases out of 5, as well as functional success, defined as a visual acuity improvement of 2 lines or more on the Snellen chart. Keeping in mind the well-known natural history of idiopathic full thickness macular holes, our preliminary results are encouraging and allow us to persevere in this surgery, whom indications and optimal methods are still to be defined. PMID- 7496575 TI - [Progressive non-inflammatory lesions of the vitreous and intermediate uveitis]. AB - A 77 year old man presented with a decreased vision, quite anterior segments, vitreous haze (++) (cells and flare) OD > OS, snow banks and snow balls at the inferior pars plana, suggesting an intermediate uveitis. Two subTenon injections of steroids were performed on the right eye, with improvement of his right visual acuity from 5 to 10/10. Cells and snow balls disappeared but the vitreous became more fluffy and more organized. Investigations did not demonstrate any systemic disease. The vitreous haze progressed bilaterally and failed to respond to systemic steroids. A year later, visual acuity was limited to hand movements (OD) and remained at 9/10 (OS). The vitreous haze + (OD) and ++ (OS) was extremely fluffy and moderately organized without inflammatory cells. Vitrectomy was performed in both eyes. Non inflammatory fibro-cellular membranes in a dense collagenous network with amorphous precipitates were observed in electron microscopy. One year later, he developed a systemic B-cell lymphoma. This unusual case illustrates the possible evolution of intermediate uveitis in a secondary PVR-like progressive non inflammatory vitreous haze with a particular increase of the amount of collagen fibrils and amorphous precipitates responsible for the fluffy aspect of the vitreous failing to respond to steroids. PMID- 7496577 TI - [Post-varicella acute retinal necrosis]. AB - A man aged 43 in good health complaints of sudden blurred vision in his right eye, 12 days after a generalized chickenpox eruption. Examination shows an intraocular inflammation with retinal necrosis in temporal periphery. The serum antibodies against varicella-zoster are positive for the IgM and IgG, confirming a recent infection by varicella zoster. Bacterial serology is negative, as well as the serology for the HSV, HIV and CMV. An intraocular production of anti varicella-zoster antibodies is also found by an anterior chamber puncture (Goldmann-Witmer ratio = 1338). A general treatment by acyclovir and corticoids is started, completed by local treatment and cryocoagulation of the retinal periphery. The evolution is favorable, with recovery of the visual acuity and cicatrization of the lesions. The severity of acute retinal necrosis as a complication of a chickenpox infection usually is moderate, with a good visual prognosis as by our patient. PMID- 7496578 TI - Discrepancy between indocyanine green and fluorescein angiography in occult choroidal neovascularization. AB - Six eyes without or with minimal evidence of choroidal neovascularization on fluorescein angiography are presented. All six eyes have marked retinal pigment epithelial alterations. Indocyanine green angiography reveals extensive hyperfluorescence, indicative of choroidal neovascularization undermining the macula in these six eyes. It is surprising that three eyes are asymptomatic and two others have a relative good visual acuity. It is suggested that hyperfluorescence observed by indocyanine green angiography without or with minimal late dye diffusion on fluorescein angiography could indicate a slow progression and thus a better prognosis of the maculopathy. PMID- 7496580 TI - Sneddon's syndrome with bilateral peripheral retinal neovascularization. AB - A 27-year old woman is presented with a five year old history of Sneddon's syndrome in whom recently peripheral retinal neovascularization in both eyes was discovered. Sneddon's syndrome is a rare clinical entity first described in 1965 and characterized by typical skin lesions (livedo reticularis) and cerebrovascular lesions occurring at early age. Central retinal artery occlusions have been reported twice in Sneddon-patients, but peripheral retinal capillary occlusions and neovascularization have not yet been reported. The role of the antiphospholipid antibodies in this association is discussed. PMID- 7496579 TI - [Posterior capsule and retinal detachment]. AB - Comparison of the incidence of retinal detachment after intracapsular and extracapsular lens extraction. PMID- 7496581 TI - Electroretinographic findings versus fluorescein angiographic appearance of the retina in patients with ischaemic and non-ischaemic central retinal vein occlusion. AB - We compared the fluorescein angiographic and electroretinographic findings in 39 eyes of 37 patients with a central retinal vein occlusion. We proved that the flash-electroretinogram is a clinically useful test to differentiate the ischaemic type of central retinal vein occlusion from the non-ischaemic type. PMID- 7496582 TI - Choroidal biopsy in the diagnosis of a suspect intraocular lesions. AB - A thirty-eight year-old man, treated medically since 1985 for a chronic pancreatitis, showed a choroidal infiltrate in the superior mid periphery of the left fundus. A thorough systemic examination could not reveal an underlying cause. The differential diagnosis of the lesion included metastasis, intraocular lymphoma and sarcoidosis. Two months later the lesion had increased both on fundoscopy and echography and was accompanied by a serous macular detachment. A choroidal biopsy showed a moderately well differentiated mucinous adenocarcinoma. The primary site could not be determined. The mucinous character is rather suggestive for a gastrointestinal origin. Gastro intestinal choroidal metastases, and more specifically the pancreatic ones, are however rare. PMID- 7496583 TI - Pseudoglioma: a clinico-pathological report. AB - The term "pseudoglioma" designates a heterogeneous group of diseases that may be confused clinically with retinoblastoma. The authors report the case of a one year old boy with leucocoria and exotropia of the left eye. Clinical examination disclosed a retrolental whitish vascularized mass. As a retinoblastoma could not be excluded with certainty, the blind eye was enucleated. Pathological examination showed a retinal vascular anomaly compatible with Coats' disease. The authors discuss the differential diagnosis of leucocoria and pseudoglioma. PMID- 7496584 TI - Predicting conditional probability distributions: a connectionist approach. AB - Most traditional prediction techniques deliver a single point, usually the mean of a probability distribution. For multimodal processes, instead of predicting the mean, it is important to predict the full distribution. This article presents a new connectionist method to predict the conditional probability distribution in response to an input. The main idea is to transform the problem from a regression problem to a classification problem. The conditional probability distribution network can perform both direct predictions and iterated predictions, the latter task being specific for time series problems. We compare this new method to fuzzy logic and discuss important differences, and also demonstrate the architecture on two time series. The first is the benchmark laser series used in the Santa Fe competition, a deterministic chaotic system. The second is a time series from a Markov process which exhibits structure on two time scales. The network produces multimodal predictions for this series. We compare the predictions of the network with a nearest-neighbor predictor and find that the conditional probability network is more than twice as likely a model. PMID- 7496585 TI - Using the Potts glass for solving the clustering problem. AB - We present an application of a Potts glass to the clustering problem. Simulated annealing in the mean field approximation is used in order to avoid local minima. The resulting updating equations are completely parallel, and very easy to implement. The model has no free parameters except for the annealing parameters. We show how the model can be implemented for some special clustering problems. The T-->0 limit of the Potts glass is identical to the vector quantization algorithm with certain increments. A comparative study of the Potts glass and vector quantization is also made, and it is shown that for difficult clustering problems, the Potts glass is far better than vector quantization. PMID- 7496586 TI - Ensemble competitive learning neural networks with reduced input dimension. AB - Conventional neural networks utilize all the dimensions of the original input patterns for training and classification. However, a particular attribute of the input patterns does not necessarily contribute to classification and may even cause misclassification in certain cases. A new ensemble competitive learning method using the reduced input dimension is proposed. In contrast to the previous ensemble neural networks which adjust learning parameters, the proposed method takes advantage of the information in each dimension of the input patterns. Since the degree of contribution of each attribute to classification is not known beforehand, the different input data sets with one dimension reduced are presented to multiple neural networks. The classification information from each competitive learning neural network is then combined to make a final decision for classification. In order to improve classification accuracy, the ambiguous output neurons are eliminated which cannot be assigned to any class after training. We use three consensus schemes to judge the classification using ensemble neural networks. The experimental results with remote sensing and speech data indicate the improved performance of the proposed method. PMID- 7496587 TI - Dynamical recurrent neural networks--towards environmental time series prediction. AB - Dynamical Recurrent Neural Networks (DRNN) (Aussem 1995a) are a class of fully recurrent networks obtained by modeling synapses as autoregressive filters. By virtue of their internal dynamic, these networks approximate the underlying law governing the time series by a system of nonlinear difference equations of internal variables. They therefore provide history-sensitive forecasts without having to be explicitly fed with external memory. The model is trained by a local and recursive error propagation algorithm called temporal-recurrent backpropagation. The efficiency of the procedure benefits from the exponential decay of the gradient terms backpropagated through the adjoint network. We assess the predictive ability of the DRNN model with meterological and astronomical time series recorded around the candidate observation sites for the future VLT telescope. The hope is that reliable environmental forecasts provided with the model will allow the modern telescopes to be preset, a few hours in advance, in the most suited instrumental mode. In this perspective, the model is first appraised on precipitation measurements with traditional nonlinear AR and ARMA techniques using feedforward networks. Then we tackle a complex problem, namely the prediction of astronomical seeing, known to be a very erratic time series. A fuzzy coding approach is used to reduce the complexity of the underlying laws governing the seeing. Then, a fuzzy correspondence analysis is carried out to explore the internal relationships in the data. Based on a carefully selected set of meteorological variables at the same time-point, a nonlinear multiple regression, termed nowcasting (Murtagh et al. 1993, 1995), is carried out on the fuzzily coded seeing records. The DRNN is shown to outperform the fuzzy k-nearest neighbors method. PMID- 7496588 TI - On the functional equivalence of fuzzy inference systems and spline-based networks. AB - The conditions under which spline-based networks are functionally equivalent to the Takagi-Sugeno-model of fuzzy inference are formally established. We consider a generalized form of basis function network whose basis functions are splines. The result admits a wide range of fuzzy membership functions which are commonly encountered in fuzzy systems design. We use the theoretical background of functional equivalence to develop a hybrid fuzzy-spline net for inverse dynamic modeling of a hydraulically driven robot manipulator. PMID- 7496589 TI - Fuzzy neural networks: between functional equivalence and applicability. AB - Research in fuzzy neural networks, which started from application oriented fuzzy system tuning, then moving to the automatic generation of fuzzy systems from data, is reaching a more mature stage, especially after the proof of functional equivalence of certain fuzzy models and neural networks. It is essential that the applicability of such developments is explored emphasizing the directions that research should follow. It can be shown that the nearest prototype classifier is functionally equivalent to an alternative fuzzy classifier model. Efficient, hardware friendly training algorithms are developed for dynamic generation of an optimum number of nearest prototypes for neural classifiers which enable the generation of fuzzy systems in real time. These systems are tested with complex applications showing the simulation results. PMID- 7496590 TI - A perspective and critique of adaptive neurofuzzy systems used for modelling and control applications. AB - This paper outlines some of the theoretical and practical developments being made in neurofuzzy systems. As the name suggests, neurofuzzy networks were developed by fusing the ideas that originated in the fields of neural and fuzzy systems. A neurofuzzy network attempts to combine the transparent, linguistic, symbolic representation associated with fuzzy logic with the architecture and learning rules commonly used in neural networks. These hybrid structures have both a qualitative and a quantitative interpretation and can overcome some of the difficulties associated with solely neural algorithms which can usually be regarded as black box mappings, and with fuzzy systems where few modelling and learning theories existed. Both B-spline and Gaussian Radial Basis Function networks can be regarded as neurofuzzy systems and soft inductive learning algorithms can be used to extract unknown, qualitative information about the relationships contained in the training data. In a similar manner, qualitative rules or information about the network's structure can be used to initialise the system. These areas, coupled with the extensive work being carried out on theoretically analysing their modelling, convergence and stability properties means that this research topic is highly applicable in "intelligent" modelling and control problems. Apart from outlining this work, the paper also discusses a wide variety of open research questions and suggests areas where new efforts may be fruitfully applied. PMID- 7496592 TI - Adenomyoepithelioma of the breast associated with low-grade adenosquamous and sarcomatoid carcinomas. AB - Six cases of invasive breast carcinoma with unusual morphological features are reported. The ages of the female patients ranged from 46 to 79 years (mean 60.5). All tumours had areas typical of an adenomyoepithelioma. In three cases adenomyoepithelioma gradually merged with low-grade adenosquamous carcinoma. In the other three patients a sarcomatoid carcinoma was associated with adenomyoepithelial areas. A common origin is proposed for these neoplasms, which extends the morphological spectrum of epi-myoepithelial cell tumours. PMID- 7496593 TI - Simple mucin-type carbohydrate antigens (T, sialosyl-T, Tn and sialosyl-Tn) in breast carcinogenesis. AB - Immunohistochemical analysis of the expression of simple mucin-type carbohydrate antigens (Tn, sialyl-Tn and T) was performed in a series of 43 cases of intraductal hyperplasia without atypia, 9 cases of intraductal hyperplasia with atypia, 54 cases of ductal carcinoma in situ (DCIS) and 26 cases of invasive breast carcinoma. We also studied 36 cases of isolated breast normal epithelium, 20 cases of "normal" breast epithelium adjacent to neoplasms and 14 cases of apocrine metaplasia. All antigens were detected in different frequencies in normal, hyperplastic, metaplastic and neoplastic breast epithelium. Tn and sialyl Tn are expressed more frequently in malignant than in benign breast epithelium; while Tn expression increases from normal to invasive carcinomas, sialyl-Tn increases until DCIS and drops in invasive carcinomas, suggesting that either there is a failure of a proportion of DCIS to progress to invasive carcinoma or loss of expression of sialyl-Tn when some carcinomas become invasive. The high frequency of Tn and sialyl-Tn expression in breast intraductal proliferations probably reflects incomplete glycosylation in these lesions, which is a well known tumour-associated phenomenon and supports the assumption that such lesions are putative precursors of breast cancer. T antigen was expressed in all groups studied, but its prevalence differed significantly between normal and neoplastic epithelium. The expression of these antigens in epithelium adjacent to carcinomas is similar to that found in isolated normal breast epithelium, whereas apocrine metaplasia has a pattern of simple mucin-type glycosylation that is specific and distinct from that of the normal breast epithelium, with a high frequency of marked expression of Tn and sialyl-Tn. The similarity of the pattern of expression of simple mucin-type antigens in metaplasia and malignant neoplasia reduces the usefulness of these markers from a diagnostic standpoint. PMID- 7496591 TI - The clinical significance of p53 aberrations in human tumours. AB - p53 aberrations are the most common genetic alteration found in human tumours and this review summarizes the current understanding of the clinical significance of p53 abnormalities. Immunohistochemical and molecular techniques can demonstrate alterations at the protein and gene level, respectively, but with a significant discordance between the findings of either technique. The tumours evaluated in this review include cancers of the breast, lung, gastrointestinal tract, genitourinary tract, and others. In most cases, only data on p53 protein are available and in each of these tumour types discrepant conclusions on the clinical value of p53 abnormalities as prognostic indicators have been reached. The role of p53 in the context of anticancer adjuvant therapy has also been analysed. Experimental data suggest that p53 affects the apoptotic response to anticancer agents, but this has not yet been proven in a clinical series where this demonstration and its effect on therapy could represent one of the most important endpoints in p53 clinical research. The use of standardized techniques to evaluate p53 gene mutation and protein accumulation within controlled clinical series of patients entering prospective trials is essential to answer the many remaining questions on the clinical significance of p53 aberrations. PMID- 7496594 TI - Relationship between ERBB2 and E-cadherin expression in human breast cancer. AB - A recent in vitro study has suggested that overexpression of ERBB2 may mediate breast tumour progression and metastasis by inhibiting the transcription of the E cadherin (E-CD) gene. To test this hypothesis in human breast cancer in vivo, we studied the relationship between the expression of both molecules in 247 breast carcinomas immunohistochemically. Five ductal carcinomas in situ overexpressed ERBB2 and showed preserved E-CD expression. Forty-four of 226 infiltrating ductal carcinomas (19.47%) showed ERBB2 overexpression, and a statistically significant relationship was found between ERBB2 overexpression and high histological grade. E-CD expression was preserved in 111 cases (49.1%) and correlated with the histological grade. However, no significant relationship was found between ERBB2 and E-CD expression. None of the 16 infiltrating lobular carcinomas expressed ERBB2 or E-CD. These observations in different histological types of breast carcinoma strongly argue against a role for ERBB2 as a transcriptional regulator of E-CD expression in most human breast carcinomas in vivo. PMID- 7496595 TI - Quantitative histopathology in lymph node-negative breast cancer. Prognostic significance of mitotic counts. AB - Reliable prognostic factors are needed to improve the stratification of patients with lymph node-negative breast cancer to different therapy modalities. We investigated the prognostic value of quantitative histopathology in a retrospective study of 98 "low-risk" breast cancer patients (T1+2N0M0) with a median follow-up of 9 years. An interactive video system and stereological and morphometric techniques were used to obtain estimates of four nuclear features (mean volume, mean profile area, volume fraction, and profile density), and two mitotic counts [mitotic profile frequency (MF) and mitotic profile density (MD)]. All measurements were performed in fields of vision sampled systematically from the whole tumour area of a routine histological section. Histological grade, histological type, and oestrogen receptor (ER) status was reassessed, whereas tumour diameter and age at diagnosis were recorded from the files. We found that all quantitative histopathological variables and ER status were correlated with histological grade. Single-factor prognostic analyses showed a highly significant value of MF (2p = 0.001) and a marginally significant value of MD (2p = 0.09), whereas no other variable approached statistical significance (2p > or = 0.25). In a multivariate Cox analysis, MF was the only parameter of significant independent prognostic value (2p = 0.03). Thus, the prognostic value of nuclear features found in previous studies could not be reproduced, whereas the marked value of mitotic counts for prediction of the outcome in patients with breast cancer was confirmed. Mitotic counts are easily obtained and may be of clinical value for identification of high-risk cases among patients with lymph node negative breast cancer. PMID- 7496596 TI - Apoptosis and cellular proliferation in oesophageal squamous cell carcinomas: differences between keratinizing and nonkeratinizing types. AB - To assess cell death and cellular proliferation activity, the apoptosis index, the Ki67 proliferative index and overexpression of p53 protein were evaluated in 69 oesophageal squamous cell carcinomas (ESCC), all surgically resected from Japanese patients. Apoptosis was examined by Gavrieli's method in histological sections, and proved to be significantly related to keratinization and ESCC progression. Overall labelling indices were 15.68 +/- 4.04 (positive/1,000 nuclei) and 6.79 +/- 0.64 respectively, in keratinizing and nonkeratinizing types. The apoptosis labelling index increased, especially in keratinizing lesions, from 4.50 +/- 0.59 with cancer invasion to mucosa through 11.46 +/- 2.70 with involvement of the submucosa up to 21.18 +/- 3.72 in cases of penetration to the muscularis propria or adventitia. The relationship between apoptosis, Ki67 scores and p53 expression was determined in identical cancer nests on serial sections. An inverse correlation was shown between the apoptosis score and the Ki67 score in both keratinizing and nonkeratinizing types. There was no significant correlation between apoptosis score and p53 expression, either overall or separately in keratinizing or nonkeratinizing types of ESCC. Our results suggest that a mechanism of induction of apoptosis similar to that operating in normal epidermis acts in keratinizing ESCC, and that as tumour volume increases, single cell death becomes more frequent. PMID- 7496597 TI - Histological grading in the deep invasive front of T1 and T2 glottic squamous cell carcinomas has high prognostic value. AB - The characteristics of the deep invasive front area of squamous cell carcinomas may reflect tumour prognosis better than other parts of the tumour. Consequently, the authors have recently developed a simple malignancy grading system based solely on the characteristics of the deep invasive front area of oral squamous cell carcinomas, which has great prognostic value. Our previous materials were somewhat heterogeneous, and the prognostic value of our system needed to be confirmed in homogeneous patient material. In the present study of 95 T1-2/N0 glottic carcinomas all treated by radiation, the high prognostic value for invasive front grading of biopsy specimens is confirmed. The grading significantly predicted local recurrence, i.e. treatment failure (P = 0.001). Histological characteristics of the deep invasive front proved to be a better indicator of prognosis than the T-category (size of tumour), and our findings may be of value in the selection of treatment. Of the individual variables in the grading system (pattern of invasion, degree of keratinization, nuclear polymorphism and host response), pattern of invasion and degree of keratinization were the strongest prognosticators in the multivariate analyses. Invasive front characteristics may also prove to be of prognostic value in other cancers. PMID- 7496598 TI - Immunohistochemical localisation of stem cell factor (SCF) with comparison of its receptor c-Kit proto-oncogene product (c-KIT) in melanocytic tumours. AB - In order to characterise the distribution and role of stem cell factor (SCF), a recently-reported growth factor for normal melanocytes, we carried out an immunohistochemical study on benign and malignant melanocytic tumours with a comparison with the presence of its receptor c-Kit proto-oncogene product (c KIT). In normal skin, SCF was mainly observed in endothelial cells of blood vessels but not frequently in basal melanocytes, whereas c-KIT was predominantly localised in tissue mast cells. In benign neoplastic melanocytes (common melanocytic naevi), localisation of SCF and c-KIT was complementary: SCF was mostly found in dermal naevus cells while c-KIT was revealed in epidermal naevus cells, although the expression of the latter antigen was not frequent. Malignant melanoma cells showed less frequent expression of these antigens than those in benign lesions. Of five cultured melanoma cell lines, SCF was observed in only one, and c-KIT was not found in any melanoma cells. No quantitative or qualitative alterations assessed by Western blot analysis were induced in the presence of phenotypic modifiers (sodium butyrate and HMBA). Present data suggest that loss of SCF expression in neoplastic melanocytes is commonly associated with malignant transformation of pigment cells rather than loss of its receptor c-KIT. PMID- 7496599 TI - Immunohistochemical detection of nm23/NDP kinase and cathepsin D in medullary carcinomas of the thyroid gland. AB - Reduced expression of nm23/NDP kinase and increased expression of cathepsin D seem to be correlated with the high metastatic potential in a variety of malignancies. The expression of nm23/NDP kinase and that of cathepsin D have been evaluated by means of an immunohistochemical technique in paraffin-embedded tissues from 44 primary medullary carcinomas of the thyroid gland (MCT) and from the corresponding lymph node metastases in 32 of these cases. In addition, lymph node metastases from 4 cases were studied. We found that 36 of 44 (82%) primary and 26 of 36 (72%) lymph node metastatic MCT were nm23/NDP kinase positive, whereas 14 of the 44 (32%) primary and 17 of the 36 (47%) lymph node metastatic MCT were cathepsin D positive. We found no indication that the nm23/NDP kinase level has any prognostic significance in MCT. The cathepsin D level is close to being prognostically significant in this study, and we cannot exclude the possibility that it could be of prognostic value. However, it seems to be quite weak, and therefore of little use in a clinical situation. PMID- 7496600 TI - Evaluation of proliferation parameters in in vivo bromodeoxyuridine labelled lung cancers. AB - In a series of 44 bronchial biopsies from patients suspected of having endobronchial lung carcinoma, the validity of proliferating cell nuclear antigen (PCNA) and Ki67 antigen as proliferative indicators was evaluated in ethanol fixed, paraffin embedded tissue. The percentages of cells positive for these markers were compared to the in vivo bromodeoxyuridine (BrdU) labelling index. A good correlation was found between PCNA immunoreactivity and BrdU labelling index, while Ki67-antigen expression showed a significant relation with BrdU labelling index and with PCNA expression. All three parameters showed a trend towards similar values for the individual cases. Based on the fact that Ki67 antigen is expressed in all cycling cells, whereas replicon-associated PCNA and BrdU only reflect the S-phase fraction, the differences between Ki67-antigen scores on the one hand and BrdU and PCNA scores on the other were smaller than expected. In order to determine the degree of concordance between immunohistochemically and flow cytometrically detected proliferation variables, BrdU incorporation was measured using both methods in duplicate bronchial specimens. Discrepancies in labelling indices were observed predominantly in DNA diploid samples, with consistently lower values in the flow cytometrically analysed specimens. In tumour specimens with an aneuploid DNA content, flow cytometric determination of proliferative activity yielded results similar to those obtained by tissue section examination. We conclude that the scores for PCNA and Ki67 antigen, immunohistochemically detected in ethanol fixed, paraffin embedded tissue reflect functional proliferative activity. PMID- 7496601 TI - Anatomical variation of the oestrogen receptor in normal myometrium. AB - Tissue from 20 surgically resected uteri was examined in order to test the hypothesis that the oestrogen receptor content of the myometrium may not be uniform throughout the uterus. All patients were premenopausal, had no history of any exogenous steroid hormone use and were in the proliferative phase of the menstrual cycle. Material was taken from the subserosal, midmyometrical and subendometrial regions of both the fundal and lower uterine segments. Care was taken to remove all endometrium. Suitable samples were analysed by radioimmunoassay (RIA) for oestrogen receptors. An adjacent block from each area was fixed in formalin and stained immunocytochemically for oestrogen receptors. The results of the RIA show a significantly higher receptor content in the subendometrial region than in either the midmyometrial or the subserosal region. No significant difference was demonstrated between the numbers of receptors in the fundus and in the lower uterine segments. The differences were also well demonstrated by immunocytochemistry, according to which 83% of all cells counted in the subendometrial region were positive, while only 61% of cells in the middle and 47% of those in the subserosal region were positive. PMID- 7496603 TI - Vinblastine and 5-fluorouracil sensitivity of xenografts of four pancreatic ductal adenocarcinomas: is there a correlation with histological and cytological tumour differentiation? AB - In a search for nuclear parameters which may predict chemosensitivity of ductal adenocarcinoma of the pancreas, the growth of four xenografted pancreatic carcinomas in response to chemotherapeutic agents was correlated with histological and cytological features of tumour differentiation. Histologically, the tumours were classified according to their ability to form glands into poorly (PaTu-2, PaTu-3), moderately (Panc-1) and well differentiated (PaTu-39) ductal adenocarcinomas. Cytologically, similar segregation of tumours was possible using the 'nuclear form factor', which was one of four nuclear parameters analysed by image cytometry on Feulgen stained tumour imprints. Histological and cytological differentiation correlated closely with tumour growth. One week after a single intraperitoneal injection of either vinblastine or 5-fluorouracil, both drugs inhibited the growth of PaTu-2 and PaTu-3 significantly. The growth of Panc-1 was only affected by vinblastine, while neither drug had an effect on PaTu-39. The results suggest that the response of pancreatic ductal adenocarcinoma to chemotherapeutic drugs may be, to some extent, predicted by histological and cytological differentiation features. However, within these lines, each tumour may show a specific response pattern. PMID- 7496602 TI - Variations in peroxisomal catalase of neonatal rat hepatocyte subpopulations. Effect of pre- and postnatal exposure to alcohol. AB - Alcohol consumption during pregnancy is teratogenic and induces severe alterations in hepatocytes. In the hepatocyte peroxisomal system, ethanol is converted in the presence of H2O2 to acetaldehyde and water. Therefore, peroxisomal catalase also acts as an antioxidant defence mechanism by removing H2O2 and preventing the formation of hydroxyl radicals in the cell. Alterations in peroxisomal catalase after pre- and pre+postnatal alcohol exposure were investigated in the rat. The effect of pre- and postnatal exposure to ethanol on hepatocyte subpopulations was analysed in isolated hepatocytes originating from periportal, intermediate and perivenous zones. Analysis of catalase revealed that the total activity and content of this enzyme were higher in 12-day-old cells than in cells from newborns and that this increment was more pronounced in treated cells. In controls, the amount of peroxisomal catalase increased mainly in periportal cells, whereas alcohol exposure induced a significant increase in the catalase of perivenous hepatocytes. We conclude that pre- and postnatal alcohol exposure mainly affects the perivenous hepatocyte peroxisomes and that the increase in peroxisomal catalase could constitute a defence mechanism against free radical generation induced by alcohol exposure during the perinatal period. PMID- 7496604 TI - Cell proliferation assessment in oncology. AB - A review of the current knowledge on cell cycle control and the techniques used to assess proliferation of normal and neoplastic cells was the focus of a workshop in Regensburg, Germany, held under the joint auspices of the Graduiertenkolleg: Therapieforschung Onkologie and the Committee on AgNOR Quantification. An overview of the recently discovered group of cyclins and their specific kinases, and of other proliferation-associated antigens, such as Ki67, PCNA and topoiseromase II alpha, was given. The topics continued with a reappraisal of modern imaging and flow-cytometric techniques. An update of the relation of AgNORs to cellular proliferation and differentiation was the link to presentations on clinical data, problems and strategies for standardization, as well as guidelines to establish the prognostic value of marker molecules. These lectures were supported by posters. Bringing together researchers from life sciences, technically oriented workers, pathologists, and clinicians resulted in a lively and constructive discussion, which is briefly summarized in the Concluding remarks. PMID- 7496605 TI - Demonstration of polyclonal origin of giant fibroadenoma of the breast. AB - We have shown that fibroadenoma of the breast is polyclonal and that phyllodes tumour is monoclonal in origin. It is not known whether a giant fibroadenoma which is histologically identical to the more usual type of fibroadenoma but grows to be a huge mass, like a phyllodes tumour, is polyclonal or monoclona. Clonal analysis was conducted on the DNA samples extracted from the paraffin sections of a giant fibroadenoma resected from a 21-year-old woman. The method used was based on trinucleotide repeat polymorphism of the X-chromosome-linked androgen receptor gene and on random inactivation of the gene by methylation. Clonal analysis showed that the giant fibroadenoma and the adjacent normal breast tissue are polyclonal in origin. Although the term giant fibroadenoma has often been used interchangeably with the term benign phyllodes tumour, because of their similarity in clinical appearance, our present results demonstrate that a giant fibroadenoma is a polyclonal fibroadenoma that has attained an immense size and is different from the monoclonal phyllodes tumour. PMID- 7496606 TI - Submandibular gland metastasis of breast carcinoma: a case report and review of the literature. AB - We report a female patient who developed a solitary metastasis in the submandibular salivary gland 2 years after primary surgery for a grade II T1 N0 M0 breast cancer. A review of the literature shows that metastases in the submandibular gland are uncommon but when they arise the site of the primary tumour is more likely to be distant than in the head and neck region. In female patients, breast tumours predominate. PMID- 7496607 TI - The hypothalamic-pituitary-adrenocortical system and sleep in man. AB - This review article summarizes the major findings about the interactions of human sleep structure and the hypothalamo-pituitary-adrenocortical (HPA) system under physiological and pathophysiological conditions, including studies that probe the sleep effects of systemically administered HPA hormones. Human sleep is regulated by a concerted action of various signal compounds acting at sleep-generating neurons whose central organization is not yet fully understood. During nocturnal sleep the endocrine system is remarkably active, the longest established finding being that growth hormone (GH) release is associated with the initiation of sleep and that there is a steep morning rise of cortisol (Weitzman et al., 1966; Takahashi et al., 1968). Moreover, the effects of exogenously administered corticosteroids and of their excessive endogenous release (e.g. Cushing's disease) were recognized more than 20 years ago. PMID- 7496608 TI - Corticotropin-releasing hormone involvement in stressor-induced alterations in sleep and in the regulation of waking. AB - Sleep responds to a variety of stressors, but the precise mechanisms whereby these alterations occur are not known. Ample evidence, however, testifies to corticotropin-releasing hormone (CRH) being uniquely situated to contribute to stressor-induced alterations in sleep. Behavioral responses to most stressors include periods of increased arousal and waking, regardless of whether the stressor is psychological in nature or results in physical insult. Furthermore, a large body of evidence suggests that CRH may also contribute to the regulation and maintenance of physiological waking. In this paper we hypothesize that CRH mediates waking, particularly after periods of exposure to acute stressors. The complex interactions of multiple systems determine the behavioral response to a particular stressor. As such, many factors determine the time course and duration of response, including stressor type, and the status of a particular system at the time of stressor presentation. We briefly review data indicating that CRH mediates physiological and behavioral responses to stressors, and present new data supporting the hypothesis that CRH may also be involved in the physiological regulation of waking. PMID- 7496609 TI - Evidence for a sleep-promoting influence of stress. AB - In the present review the data supporting the existence at the central level of a stress-sleep relation are reported and discussed. An immobilization stress of 1 or 2 hour(s) is accompanied by a marked polygraphic waking and followed by a significant sleep rebound concerning mainly paradoxical sleep (PS). During the restraint, an important release of 5-hydroxyindoles [5-OHles, a good index of serotonin (5-HT) release] occurs in the basal hypothalamus (BH). This release, produced by the nerve endings originating from the nucleus raphe dorsalis (nRD), might secondarily influence the release and/or the synthesis of hypnogenic substances directly involved in the sleep rebound production. Corticotropin-like intermediate lobe peptide (CLIP, or ACTH18-39) is a peptide possessing hypnogenic properties and derived from proopiomelanocortin (POMC) whose perikarya are contained within the BH (arcuate nucleus). The POMC nerve endings impinge on the nucleus raphe dorsalis, a structure containing sleep permissive components upon which CLIP acts to trigger sleep. It remains to be defined how the activity of the neuronal loop described above is impaired under chronic stress conditions. PMID- 7496610 TI - Influence of host defense activation on sleep in humans. AB - Despite considerable progress in our understanding of the phenomenology of sleep and wakefulness, their regulation and peculiar functions are poorly understood. Recent animal research has revealed considerable evidence for interactions between host defense and sleep. Therefore, it has been hypothesized that host response mediators, mainly cytokines like interleukin-1 (IL-1), are involved in physiological sleep regulation. Furthermore, it has been suggested that sleep, and non rapid eye movement (NREM) sleep in particular, has an immuno-supportive function. In humans, sleep-host defense interactions are just starting to be understood. There is quite good evidence that some viral diseases cause excessive sleepiness. Other infectious diseases induce, however, serious disturbances of the distribution of sleep and wakefulness rather than excessive sleep. In addition, some disorders with excessive sleep, daytime fatigue or disturbed night sleep as prominent symptoms are thought to involve, at least in part, immuno pathophysiological mechanisms. Experimental settings have only recently been used to elucidate host defense-sleep interactions in humans. The effects of endotoxin, a cell-wall lipopolysaccharide of gram-negative bacteria, on sleep have been tested in different settings in healthy volunteers. Endotoxin transiently suppresses rapid eye movement (REM) sleep independently of the time of the day of administration. Only low doses, given in the evening, promote NREM sleep. Electorencephalogram (EEG) power in higher frequency bands is enhanced during NREM sleep, whereas delta activity is not affected. In rats and rabbits, on the other hand, the effects of endotoxin and of the mediators of its activity on REM sleep are variable. Enhanced NREM sleep is a common finding and most pronounced during the active part of the nycthemeron and, in general, EEG delta activity is augmented. In view of these species differences, hypotheses regarding the underlying mechanisms and the biological significance of host defense-sleep interactions, primarily derived from the results of animal studies, may not entirely fit human physiology. They should therefore be re-evaluated and probably modified, through the use of additional experimental approaches in humans. PMID- 7496611 TI - Cytokines in sleep regulation. AB - The central thesis of this essay is that the cytokine network in brain is a key element in the humoral regulation of sleep responses to infection and in the physiological regulation of sleep. We hypothesize that many cytokines, their cellular receptors, soluble receptors, and endogenous antagonists are involved in physiological sleep regulation. The expressions of some cytokines are greatly amplified by microbial challenge. This excess cytokine production during infection induces sleep responses. The excessive sleep and wakefulness that occur at different times during the course of the infectious process results from dynamic changes in various cytokines that occur during the host's response to infectious challenge. Removal of any one somnogenic cytokine inhibits normal sleep, alters the cytokine network by changing the cytokine mix, but does not completely disrupt sleep due to the redundant nature of the cytokine network. The cytokine network operates in a paracrine/autocrine fashion and is responsive to neuronal use. Finally, cytokines elicit their somnogenic actions via endocrine and neurotransmitter systems as well as having direct effects neurons and glia. Evidence in support of these postulates is reviewed in this essay. PMID- 7496612 TI - Sleep regulation: interactions among cytokines and classical neurotransmitters. AB - The role of classical neurotransmitters in sleep regulation is amply documented (Hobson and Steriade, 1986). In recent years evidence has been gathered that immunoactive molecules, infectious agents and their components, or cytokines play some part in sleep regulation (Krueger and Obal, 1994; Opp et al., 1992; Moldofsky, 1994). Different cytokines possess hypnogenic properties when injected centrally or systemically to different animal species and their role in physiological sleep regulation is currently under investigation. Little is known of how cytokines and classical neurotransmitters interact and of the relevance of this interaction in sleep induction and maintenance. The present paper (i) reviews data on this topic; (ii) proposes a unitary interpretation whenever possible; and (iii) raises questions that might be addressed by future studies. PMID- 7496613 TI - Interactions between cytokines and nitric oxide. AB - There is now an impressive range of evidence supporting the important role of cytokines in sleep regulation (see Krueger et al., 1995; De Simoni et al., 1995). It has also been reported that inhibition of nitric oxide (NO) synthesis suppresses sleep in rabbits (Kapas et al., 1994). This is not surprising, since NO is closely involved in neurotransmission (Garthwaite, 1991; Schuman and Madison, 1994) and cytokines are the major inducers of NO synthesis (Hibbs et al., 1990). Further, it is now clear that NO plays an important role in modulating immune responses, possibly through the differential regulation of cytokine synthesis (Taylor-Robinson et al., 1994). In this article, I will provide evidence for the interactions between cytokines and nitric oxide, and discuss their implications in the regulation of immune responses. I shall illustrate these mainly with results from my coworkers and I, from our laboratory rather than attempting an exhaustive review of the subject. PMID- 7496614 TI - Prostaglandins and sleep. AB - The concept of humoral regulation of sleep was initially proposed by a French neuroscientist, Henri Pieron of Paris, in the first decade of this century. He and his associate Legendre were the first to show the presence of a sleep inducing substance in the cerebrospinal fluid (CSF) of sleep-deprived dogs. Concurrently and independently, Kuniomi Ishimori of Nagoya University, Nagoya, Japan, employing a similar experimental approach, also demonstrated a sleep inducing substance in the CSF of sleep-deprived dogs. During the next 80 years or so, more than 30 so-called endogenous sleep substances have been reported to exist in the brain by numerous investigators, CSF, and other organs and tissues of mammals. However, their physiological relevance has remained uncertain in most instances. In this review, we shall focus upon our own work concerning the molecular mechanisms of sleep-wake regulation by prostaglandins (PGs) D2 and E2, with special emphasis on the recent developments during the last several years. PMID- 7496615 TI - Towards a molecular biology of the circadian clock and sleep of mammals. AB - Behavioral states of rest and activity are temporally organized. Since the beginning of life on Earth, plants and animals have been forced to adapt to the daily rhythm of the planet's rotation about its axis. In complex vertebrates (birds and mammals), rest and activity have evolved into the electrophysiologically and behaviorally distinct states of sleep and wakefulness. The evolutionary emergence of bouts of rapid eye movement (REM) sleep may be even more recent; the echidna, one of the earliest mammals, lacks this sleep stage (Siegel et al., 1994), The cycling of these behavioral states is under neural control, and much is known about their cellular basis, but the underlying events at the molecular level are virtually unknown. Here each of us highlights some of the new approaches for investigating the molecular substrate for behavioral state control of circadian rhythmicity (WJS) and sleep (PJS) in mammals. PMID- 7496617 TI - Advances in atherosclerosis. AB - In the past few years major advances have changed the way we think about atherosclerosis. Treatments for hyperlipidaemia not only regress angiographic stenosis and reduce coronary mortality; in meta-analysis, and in large trials of secondary prevention, they appear to reduce overall mortality. Reduction in events precedes regression of stenosis, partly through stabilization of plaque, and partly by reversal of the endothelial dysfunction caused by oxidized LDL. The study of lipid disorders has been advanced by new understanding of the role of apolipoproteins and lipoprotein (a). Significant advances have also been made in the understanding of effects of angiotensin, fibrinogen, homocysteinaemia, platelets, genetics of atherosclerosis, autoimmunity, flow disturbances and wall stress. Most importantly, because of a revolution in our understanding of the endothelium, we can now begin to make sense of some of the effects of risk factors such as lipids, menopause, flow disturbances, and pressure, to begin to understand atherosclerosis, arterial remodelling and the protective effect of oestrogen in a new light. These changes will profoundly affect the way we practise vascular medicine and neurology, and will give us powerful new ways to help our patients. PMID- 7496616 TI - Sleep deprivation and human immune function. PMID- 7496618 TI - The proximal aorta: a source of stroke. AB - The availability of the sophisticated imaging techniques of transoesophageal echocardiography and epiaortic ultrasound scanning have drawn attention to the ascending aorta and aortic arch as a potential source of embolic stroke. Several studies have shown an association between atheroma in this region and cerebral ischaemic events. Although aortic atheroma is associated with vascular disease in other arterial territories, two large controlled studies have shown it to be a risk factor for stroke, independently of other major risk factors such as carotid vascular disease, cardiac disease and hypertension. In observational and case control studies, the risk of stroke is higher in the presence of certain echocardiographic appearances of atheromatous plaque--these include plaque thickness of > 4-5 mm, surface irregularity suggesting plaque ulceration and mobile elements suggesting superimposed thrombus. However, longitudinal studies are required to evaluate the prognostic significance of such findings. Several therapeutic options have been described or suggested in patients with and without stroke in whom aortic atheroma is demonstrated, but the optimal management of such patients is yet to be determined in prospective controlled trials. PMID- 7496619 TI - The genetics of cerebrovascular disease. AB - Genetics contribute to all aspects of cerebrovascular disease from the underlying pathology, through acute neuronal injury, to recovery and long-term prognosis. By understanding better the genetic contribution to the risk for cerebrovascular disease we can work toward the day when simple genetic tests can be used to assess an individual's risk for stroke (and cardiovascular disease) in the same way that we now use routine screening of blood pressure or cholesterol (Harrap, 1994b). Finally, it is worth noting that genetic effects do not work 'in vacuo'. They are subject to environmental influences which impact on the individual's phenotypic characteristics. Modification of these environmental factors may have a significant effect on the action of certain expressed genes and their putative mutations. PMID- 7496620 TI - Intracerebral haemorrhage. AB - Intracerebral haemorrhage accounts for 15% of strokes. Its mechanisms include hypertension, cerebral amyloid angiopathy, rupture of vascular malformations, bleeding into primary or metastatic brain tumours, coagulopathies (due to the use of anticoagulants and thrombolytic agents), sympathomimetic drug effect (amphetamines, phenylpropanolamine, and cocaine), and vasculitis. The clinical presentation reflects both the general effects of increased intracranial pressure, and the neurological deficits that result from the specific location of the haemorrhage. Its diagnosis is based on computerized tomography, which identifies haemorrhage as a high-attenuation mass within the brain substance, and magnetic resonance imaging, which in addition estimates the age of the haemorrhage by identifying sequential patterns of transformation of the haemoglobin molecule within the haematoma. The mortality in intracerebral haemorrhage is dependent on the size and location of the haematoma. A reliable clinical parameter for the prediction of outcome is the Glasgow Coma Scale score at presentation. The management of intracerebral haemorrhage involves: (a) the prevention and treatment of increased intracranial pressure; and (b) the choice between surgical and nonsurgical treatment, a clinical decision that is still controversial as a result of the paucity of controlled clinical data comparing both treatment modalities. PMID- 7496621 TI - The penumbra, therapeutic time window and acute ischaemic stroke. AB - There is a great deal of evidence that an ischaemic penumbra exists in animals and humans after the occurrence of focal brain ischaemia (Hossmann, 1994). The concept of the penumbra leads to the idea of a therapeutic time window. Because, if the region of irreversible injury (infarction) after focal ischaemia evolves in time and space, then the possibility of therapy to interfere becomes a tenable hypothesis. All of the acute stroke therapies given after onset have their basis from this hypothesis of a therapeutic time window (Fisher, 1995). As previously alluded to, a more apt term might be a window-shade, because this metaphor suggests a more dynamic event. The time and location of potentially salvageable ischaemic brain tissue after ischaemic stroke is a moving target and many unanswered questions remain. The data from animal stroke models support 2-3 hours as the time when intervention is likely to be beneficial in rats. Non-human primate data are scarce, but the few studies available do imply that at 3-4 hours after stroke onset some ischaemic tissue remains potentially salvageable. In humans, we really do not know what the time window is and we must remember that it is likely to be highly variable among individuals. This variability relates to many factors including the status of collateral flow, patient age, coexistent metabolic abnormalities (i.e. hyperglycaemia), premorbid medications and many other confounding variables. All acute stroke intervention trials are trying to initiate therapy within 6-8 hours after onset and the earlier, presumably the better. However, this approach is based upon population averaging, since we have had no convenient and reliable mechanism to determine, if an individual patient has viable tissue when therapy can be started. The availability of an imaging modality that could distinguish the presence and extent of salvageable ischaemic tissue would greatly facilitate stroke therapy trials and ultimately the selection of patients when proven therapies are available. The new MRI techniques might afford this possibility. As we enter the exciting era of effective therapy for acute ischaemic stroke, the issues surrounding the therapeutic time window(shade) will become more critical, because it is this critical time that will define the success or failure of our interventions. Therefore it is incumbent upon basic stroke researchers and clinicians to continue to define the ischaemic penumbra and to develop readily applicable mechanisms to identify and treat this moving target. PMID- 7496622 TI - Interventional neuroradiology. AB - Interventional neuroradiology is a dynamic subspecialty which is rapidly gaining new applications in the treatment of neurological diseases. The most common therapeutic role is still in the treatment of brain AVMs, aneurysms and fistulae in close association with neurosurgical support. Endovascular treatment of aneurysms with thrombogenic coils is a promising technique which can be performed with low morbidity. The goal of complete aneurysm occlusion, including the neck, has not yet been achieved in all cases and the procedure has not been shown to possess clear advantages over surgical clipping. Pre-operative embolization of tumours and spinal lesions are common procedures in some centres. Angioplasty and papaverine infusion for treatment of cerebral vasospasm are being performed in most neurosurgical teaching centres. Although angiographic results are often impressive, corresponding clinical improvement is not always seen, particularly when treatment is delayed. Thrombolysis for treatment of acute stroke and angioplasty for treatment of cerebrovascular atherosclerosis are new techniques which await scientific validation before being accepted as standard therapies. PMID- 7496623 TI - Recovery from stroke: rehabilitation. AB - Most of the neurological and functional recovery following stroke occurs during the first 3-6 months, although some patients show prolonged and unexpected levels of recovery. Research shows that the determination of outcome is multifactorial and includes demographics and psychosocial as well as medical factors. New information on how the brain recovers from injury has given us insights into which therapy approaches theoretically might be beneficial. In some cases recovery may be facilitated by the interaction of appropriate therapies and pharmacological interventions. Comprehensive rehabilitation programmes and integrated acute care rehabilitation stroke units appear to produce improved functional recovery over the standard of care. The reasons are unclear, but may include coordination and standardization of care as well as the early start of rehabilitation. Future research should continue to enhance the mechanisms of recovery from stroke and the approaches to facilitating the recovery process. PMID- 7496624 TI - Carotid endarterectomy: emerging indications. AB - Several randomized trials have recently reported on the benefit of carotid endarterectomy for symptomatic or asymptomatic patients. A positive benefit has been found for symptomatic patients with > or = 70% stenosis measured by angiogram with a particular formula and performed with high surgical skill. Symptomatic patients with < 70% stenosis continue to be randomized and followed. The answer for this group is expected in the next two years. The randomized trials of asymptomatic patients have not established the benefit for carotid endarterectomy clearly. The most recent trial shows the absolute risk reduction at 5 years is only 5.9% or less than 1.5% risk reduction per year. A fifth trial continues in Europe. PMID- 7496625 TI - Vascular cognitive impairment: a new approach to vascular dementia. AB - Vascular dementia has remained a confused concept since its origin over a century ago. Recently developed criteria, which have not met with universal acceptance, erroneously base their description of vascular dementia on Alzheimer's disease, an error that is founded in the historical confusion of the two conditions. These errors include requirements for prominent memory loss and the occurrence of a substantial degree of cognitive impairment before dementia can be diagnosed. They also treat vascular dementia as a single condition and fail to address the question of aetiology. Furthermore, these criteria largely ignore such data as is already available regarding vascular dementia. Vascular dementia is preventable. Thus cases need to be detected as soon as possible to avoid unnecessary deterioration. To correct these errors a new concept, that of vascular cognitive impairment, is proposed. This concept seeks to identify cognitive impairment due to cerebrovascular disease at the very earliest stage and, by identifying the aetiology, enable the institution of appropriate preventive therapy. PMID- 7496627 TI - Xenopus F-cadherin, a novel member of the cadherin family of cell adhesion molecules, is expressed at boundaries in the neural tube. AB - Development of the vertebrate CNS begins during neurulation when the neural plate gives rise to the neural tube. During neurulation, the different regions of the CNS can be identified in part by the appearance of flexures in the walls of the neural tube. Here we report the isolation and characterization of F-cadherin; a novel member of the cadherin family of cell adhesion molecules, which is expressed at flexures in the neural tube of Xenopus embryos. We show that F cadherin is first expressed at neural plate stages, that its expression is altered when patterning of the neural tube is perturbed, and that its expression marks boundaries in the neural tube where cell mixing is restricted. These observations suggest that F-cadherin contributes to the regionalization of the neural tube perhaps by mediating differential cell adhesion. PMID- 7496626 TI - Retinoic acid-mediated increase in TrkA expression is sufficient to elicit NGF dependent survival of sympathetic neurons. AB - Sympathetic neurons depend on the classical neurotrophin NGF for survival by the time they innervate their targets, but the mechanisms controlling the onset of NGF responsiveness in developing neuroblasts have not been defined. Immature chick sympathetic neurons are unresponsive to NGF, but express low mRNA levels of the high-affinity NGF receptor trkA. Treatment with retinoic acid (RA) leads to increased levels of both trkA mRNA and protein, a response mediated through retinoic acid receptor alpha (RAR alpha). Ectopic expression of trkA in these cells results in the ability to survive with NGF, suggesting that RA-induced trkA expression is sufficient to elicit NGF-dependent survival. Our data establish a mechanism controlling NGF responsiveness and implicate a function for RA at defined late stages of neuron development. PMID- 7496628 TI - Premature Schwann cell differentiation and hypermyelination in mice expressing a targeted antagonist of the POU transcription factor SCIP. AB - The transcription factor SCIP is expressed by immature neurons and Schwann cells of the developing central and peripheral nervous systems, but this expression is largely extinguished when these cells fully differentiate. In immature Schwann cells in vitro, SCIP acts as a repressor of the myelin-specific genes that mark full differentiation. We have generated transgenic mice that express a dominant negative antagonist of SCIP, specifically targeted to developing Schwann cells. This antagonist--designated delta SCIP--is transcriptionally inactive, but retains full DNA-binding activity. Mice that express delta SCIP exhibit a debilitating peripheral neuropathy that results from developmentally advanced Schwann cell differentiation, over-expression of myelin-specific gene products, and hypermyelination. These results suggest that SCIP functions as a transcriptional sensor of differentiation cues and thereby regulates the time and place at which Schwann cells differentiate. PMID- 7496629 TI - Local control of granule cell generation by cerebellar Purkinje cells. AB - Cerebellar Purkinje cells were ablated by the specific expression of diphtheria toxin in these cells in transgenic mice. Purkinje cell degeneration during early postnatal development shows a zonally restricted pattern which has been exploited in order to look for local secondary effects. The most obvious early effect is the alignment of gaps in the Purkinje cell layer with dramatically thinned zones in the overlying EGL, the germinal layer from which granule cells are generated. Within these EGL zones in the transgenic mutant, markers that distinguish matrix from mantle cells demonstrate a preferential loss of the proliferative cells. Comparison of BrdU incorporation in the mutant vs wild-type confirms the reduction in proliferation. In the mutant, in situ labeling of DNA fragmentation associated with apoptotic cell death shows abundant labeling of granule cells that have exited the EGL, but not of progenitor cells in the EGL. Thus, although a trophic role for Purkinje cells has been well documented, these observations further suggest a mitogenic role which can be exerted locally. PMID- 7496630 TI - Patterns of presynaptic gene expression define two stages of synaptic differentiation. AB - To elucidate the sequence of molecular events leading to nerve terminal differentiation, we have examined the regulation of expression of presynaptic protein genes during synapse formation in vivo. In the chick ciliary ganglion (CG), synaptophysin IIa and synaptophysin IIb mRNAs showed threefold increases relative to neurofilament-M mRNA during the time of target contact [Embryonic Day 7 (E7)-E9]. Expression of synaptotagmin I mRNA also increased severalfold over this time interval. Thus, mRNAs for three synaptic vesicle proteins are upregulated coordinately during synaptogenesis. In contrast, the major increase in choline acetyltransferase (ChAT) mRNA (four- to fivefold) occurred between E15 and E20, coincident with the maturation of synapses in the CG. Coincident with ChAT upregulation, there is a switch in the relative abundance of mRNAs encoding vesicle protein isoforms. In particular, mRNAs encoding synaptophysin IIb and synaptotagmin II (which is undetectable at E9) become predominant. Therefore, although synaptic vesicle protein mRNAs are upregulated in a first phase of differentiation at the time of synapse formation, a temporally distinct phase of presynaptic protein gene regulation, associated with the specific maturation of synapses, is also apparent. PMID- 7496631 TI - The glypiated neuronal cell adhesion molecule contactin/F11 complexes with src family protein tyrosine kinase Fyn. AB - Glycosyl phosphatidylinositol-anchored glycoproteins of the immunoglobulin superfamily play an important role in the formation of neuronal networks during development. The mechanism whereby neuronal GPI-linked molecules transduce recognition signals remains to be established. Analysis of detergent-resistant immune-complexes reveals that the glypiated neuronal cell adhesion molecule contactin/F11 specifically complexes with the cytoplasmic, nonreceptor type src family tyrosine kinase Fyn. Antibody-mediated cross-linking of contactin/F11 on embryonic chick neuronal cells leads to an increase of the Fyn-activity coprecipitated with contactin/F11, and elevates phosphorylation of an additional 75/80 K component within the contactin/F11-immune-complex. Additionally, binding of ligands, i.e., contactin/F11-specific antibody or tenascin-R, a natural ligand of contactin/F11, to the surface of HeLa transfectants expressing contactin/F11, causes capping of contactin/F11 and a concomitant change in the distribution of the intracellular kinase Fyn, thus confirming their physical association. This indicates that contactin/F11-mediated signaling requires Fyn. PMID- 7496632 TI - Identification by differential RT-PCR of a novel paired homeodomain protein specifically expressed in sensory neurons and a subset of their CNS targets. AB - Sensory neurons are a major derivative of the neural crest for which there have been no definitive molecular markers in mammals. We have developed a method that combines differential hybridization with degenerate RT-PCR to rapidly screen gene families for members exhibiting differential expression among tissues or cell types. We used this approach to search for transcription factor-encoding genes specifically expressed in mammalian sensory neurons. A novel paired homeodomain protein, called DRG11, was identified. DRG11 is expressed in most sensory neurons, including trkA-expressing neurons, but not in glia or sympathetic neurons. Unexpectedly, it is also expressed in the dorsal horn of the spinal cord, a region to which NGF-dependent sensory neurons project. These data suggest that DRG11 is not only a useful marker for sensory neurons, but may also function in the establishment or maintenance of connectivity between some of these neurons and their central nervous system targets. PMID- 7496633 TI - Protein masking of a ribosomal RNA epitope is an early event in afferent deprivation-induced neuronal death. AB - Cell death in the developing nervous system is regulated by both afferent synaptic activity and target-derived neurotrophic factors. Loss of afferent innervation via unilateral cochlea removal results in the death of 20-40% of neurons in the neonatal chick cochlear nucleus, nucleus magnocellularis (NM). The process of NM neuronal death involves structural and functional alterations in ribosomes, including decreased protein synthesis, loss of immunoreactivity for a monoclonal anti-ribosomal RNA (rRNA) antibody, Y10B, and eventual ribosome degradation. In the present report we confirm that the Y10B antibody binds specifically to ribosomes in chick NM neurons by electron microscopy. We then performed experiments designed to determine whether loss of rRNA immunoreactivity observed in NM neurons following cochlea removal involves induction of a protein rRNA interaction. Brain stem tissue from animals subjected to unilateral cochlea removal was treated with protease prior to immunolabeling. Protease treatment restored rRNA immunoreactivity after 3 h of afferent deprivation, confirming that afferent deprivation induces protein-rRNA interactions which mask the Y10B epitope. Immunoprecipitation experiments confirmed that the Y10B antibody recognizes a specific rRNA sequence without posttranscriptional modification. PMID- 7496634 TI - Psychiatry and philosophy. PMID- 7496635 TI - Psychological treatments for depression. An update. PMID- 7496636 TI - Aspects of oxidative metabolism in schizophrenia. PMID- 7496637 TI - In vivo magnetic resonance spectroscopy. Applications in psychiatry. AB - BACKGROUND: Nuclear magnetic resonance is a non-destructive and non-invasive technology that is highly suited for research in psychiatry. It is establishing itself as a versatile means of studying brain morphology, chemistry and function and is finding a place in the diagnosis of disease, monitoring of treatment and the study of basic brain processes. METHOD: A literature review was undertaken. RESULTS: Magnetic resonance spectroscopy has been shown to distinguish between psychiatric disorders, and has provided evidence of their pathophysiological mechanisms. CONCLUSIONS: Spectroscopy in particular opens a window, for the first time, on the study of in vivo brain chemistry. PMID- 7496638 TI - Treatment of sexual dysfunctions by sex therapy and other approaches. AB - BACKGROUND: The treatment of sexual dysfunctions underwent a great change when sex therapy was developed more than 25 years ago. Since then the treatment programme has been modified in various ways, the response to treatment evaluated and other treatment approaches introduced. METHOD: A review of the literature concerning the application and outcome of sex therapy and other treatments for sexual dysfunction was conducted. RESULTS: The format of effective conjoint sex therapy is now fairly clear and there is good understanding of the sexual dysfunctions that respond best to this treatment and the couples most likely to benefit. Less is known about the effects of treatment of individuals without partners, bibliotherapy and combining sex therapy with marital therapy and with physical methods of treatment. CONCLUSIONS: Sex therapy is now a well-established form of treatment. It should be more widely available for patients seen in psychiatry departments. PMID- 7496639 TI - A 15-20 year follow-up of adult psychiatric patients. Psychiatric disorder and social functioning. AB - BACKGROUND: An exploratory study was undertaken of the importance of personality disorder in predicting the long-term outcome for both episodic disorders and social functioning. METHOD: In 1966-67, a representative series of patients with children, free of episodic illness for at least one year, was sampled from the Camberwell Psychiatric Register and systematically assessed over a four-year period, using measures of known reliability and validity. Psychiatric disorder was measured using a PSE-compatible instrument. The follow-up after 15-20 years used the PSE and a systematic assessment of social functioning. RESULTS: Overall outcomes were similar across diagnoses, but an initial categorical diagnosis of personality disorder predicted much poorer outcomes on psychiatric and social measures for patients with unipolar depressive disorders than for those with other diagnoses. CONCLUSIONS: The findings indicate the importance for prognosis of including a systematic assessment of personality disorder in the clinical assessment of patients with depressive disorder. PMID- 7496640 TI - The Edinburgh Primary Care Depression Study: personality disorder and outcome. AB - BACKGROUND: Little is known about the impact of personality pathology on the treatment outcome of major depressive illness in primary care in the UK. METHOD: Patients meeting criteria for DSM-III major depressive disorder were randomly allocated to one of four treatments each lasting 16 weeks, then followed up for 18 months. Assessments were made of depressive symptoms, personality and social functioning. Personality was assessed at maximum improvement or 16 weeks. RESULTS: The prevalence of personality disorder (PD) in the sample of 113 patients was 26%. Patients with a PD were significantly younger and rated more depressed at entry than patients with no personality disorder (NoPD). On completion of treatment patients with a PD were significantly more depressed and had poorer social functioning than the NoPD group. After 18 months there were no differences in ratings of depression or social functioning between the groups. CONCLUSIONS: There was substantial improvement in both the PD and NoPD groups. The presence of personality pathology delays recovery from major depressive illness. PMID- 7496641 TI - The Maine and Vermont three-decade studies of serious mental illness. I. Matched comparison of cross-sectional outcome. AB - BACKGROUND: This study compared long-term outcome of serious mental illness in two states using a matched design to assess psychiatric rehabilitation programmes; Vermont subjects participated in a model psychiatric rehabilitation programme, while the Maine group received more traditional care. METHOD: Maine and Vermont subjects (n = 269) were matched by age, sex, diagnosis, and chronicity. Demographic, illness, and life history information were abstracted from hospital records by clinicians blind to outcome. DSM-III criteria were applied retrospectively. Outcome was assessed by clinicians blind to history. RESULTS: Vermont subjects alive at follow-up (n = 180) were more productive (P < 0.0009), had fewer symptoms (P < 0.002), better community adjustment (P < 0.001) and global functioning (P < 0.001) than Maine subjects (n = 119). CONCLUSIONS: Outcome differences may be due to Vermont's model programme and a policy of allowing an earlier opportunity for community life. PMID- 7496642 TI - The Maine and Vermont three-decade studies of serious mental illness. II. Longitudinal course comparisons. AB - BACKGROUND: This paper supplements the cross-sectional outcome comparisons of the companion paper by providing a brief account of the longitudinal courses of the Maine and Vermont samples across several outcome domains. METHOD: A Life Chart method was used to document changes in individual lives over the domains of residence, work, income source, and use of community resources over a 20-year period. Reliability studies between states were conducted. RESULTS: Throughout much of the period, more Vermont subjects lived independently, were working, and were less likely to use community resources compared to Maine subjects. CONCLUSIONS: Differences in both policies and programmes contributed to course differences between the groups. System characteristics that may lead to better outcomes are discussed. PMID- 7496643 TI - Regional brain activity in chronic schizophrenic patients during the performance of a verbal fluency task. AB - BACKGROUND: This study examined the pattern of cerebral blood flow observed in chronic schizophrenic patients while they performed a paced verbal fluency task. Such tasks engage a distributed brain system associated with willed action. Since willed action is impaired in many chronic schizophrenic patients we hypothesised that task performance would be associated with an abnormal pattern of blood flow. METHOD: Positron emission tomography (PET) was applied to 18 chronic schizophrenic patients stratified into three groups on the basis of verbal fluency performance and current symptoms. Regional cerebral blood flow (rCBF) was measured while the patients performed (a) verbal fluency, (b) word categorisation, and (c) word repetition. Results were compared with six normal controls matched for age, sex and premorbid IQ. Analysis was restricted to six brain regions previously identified in studies of normal volunteers. RESULTS: In five brain areas, including the left dorsolateral prefrontal cortex, the patients showed the same pattern of activation as control subjects. However, in the left superior temporal cortex, all patient groups failed to show the normal decrease in blood flow when verbal fluency was compared with word repetition. CONCLUSION: These observations suggest that (a) chronic schizophrenic patients can show a normal magnitude of frontal activation when matched for performance with controls, and (b) they fail to show the expected reductions of activity in the superior temporal cortex. This latter result may reflect abnormal functional connectivity between frontal and temporal cortex. PMID- 7496644 TI - Somatic delusions in schizophrenia and the affective psychoses. AB - BACKGROUND: Delusions relating to the body, a ready source of information about the immediate experiences of psychotic patients, have not been systematically studied. We attempted an account of the phenomena, looking for differences between diagnostic groupings in the type and lateralisation of such phenomena, and for evidence of localisation. METHOD: Somatic delusions elicited at interview with 550 Research Diagnostic Criteria-diagnosed psychotic patients were categorised according to content, and the results were compared across diagnostic groupings. RESULTS: Significant differences were demonstrated, both at the level of individual delusions and in the nature and overall pattern of such delusions. There were also differences between diagnostic groups in the choice of body parts involved. Among male patients there were significant differences in laterality between the groups, with schizophrenic subjects locating abnormal phenomena principally on the left and depressive subjects on the right. A provisional taxonomy of bodily delusions was developed. CONCLUSION: Phenomenological differences between the psychoses were demonstrated and the results offer some support for current hypotheses of localisation of brain dysfunction in the psychotic illnesses. PMID- 7496645 TI - Child and adolescent psychiatric presentations of second-generation Afro Caribbeans in Britain. AB - BACKGROUND: A clinical sample was used to investigate whether second-generation Afro-Caribbean children differed from other British-born children in their psychiatric presentation or vulnerability to risk factors. METHOD: Second generation Afro-Caribbean patients (n = 292) were compared with a predominantly white group of patients (n = 1311) who lived in the same inner-city area and attended the same child psychiatric clinic between 1973 and 1989. Data on psychiatric presentation and background factors were systematically recorded at the time of the initial clinical assessment. RESULTS: Afro-Caribbean patients were exposed to more socio-economic disadvantage but less family dysfunction. The ratio of emotional to conduct disorders was lower among Afro-Caribbean than among the comparison patients--an effect that was not evidently due to demographic factors or diagnostic bias. Most risk factors for emotional or conduct disorders had comparable effects on Afro-Caribbean and comparison patients. Psychotic and autistic disorders were disproportionately common among the Afro-Caribbean patients. CONCLUSIONS: Second-generation Afro-Caribbean children differ somewhat from other British-born children in their psychiatric presentation--a difference that has persisted over the 1970s and 1980s and that deserves more investigation than it has received to date. PMID- 7496646 TI - Effect of exposure to dothiepin and northiaden in breast milk on child development. AB - BACKGROUND: This study looks at the outcome of infants exposed to dothiepin in breast milk in an attempt to guide clinicians on the risk-benefit ratio of breast feeding when on antidepressants. METHOD: Thirty women, who had had HDRS scores > 15 within the first five years postpartum from the same women's hospital, were assessed with their children 3-5 years postpartum; half had breast-fed while on dothiepin (study group). Thirty-six non-depressed women were also assessed. Rating scales assessed depression, anxiety, self-esteem, personality, social support, marital relationship, child behaviour and temperament. The children were assessed by the McCarthy Scale. RESULTS: Comparisons of the two depressed groups showed no significant differences on any measures except marital conflict and child behaviour, which were the most disturbed in the study group (P < 0.001). Overall cognitive scores for the children did not differ between the groups. Higher levels of dothiepin and northiaden were associated with higher cognitive scores on subscales (P = 0.02). CONCLUSIONS: We are cautiously optimistic about the lack of any negative associations between cognitive development and exposure to dothiepin via breast milk. PMID- 7496647 TI - Paroxetine in the treatment of panic disorder. A randomised, double-blind, placebo-controlled study. AB - BACKGROUND: This study compared the efficacy and tolerability of paroxetine with placebo in the treatment of panic disorder. METHOD: After three weeks of placebo, patients received 12 weeks of treatment with paroxetine (20, 40, or 60 mg) or placebo, and finally two weeks of placebo. Dosages were adjusted according to efficacy and tolerability. Standardised cognitive therapy was given to all patients. The primary measure of outcome was reduction in the number of panic attacks. RESULTS: Analysis of the results showed statistically significant differences in favour of paroxetine between the two treatment groups in two out of the three primary measures of outcome, i.e. 50% reduction in total number of panic attacks and number of panic attacks reduced to one or zero over the study period. For the third measure of outcome, the mean change in the total number of attacks from baseline, there was a positive trend in favour of paroxetine. The results of the primary measures of outcome were strongly supported by the results of the secondary efficacy measures of outcome. In addition, paroxetine, at all doses, was very well tolerated. CONCLUSION: Paroxetine plus cognitive therapy was significantly more effective than placebo plus cognitive therapy in the treatment of panic disorder. PMID- 7496648 TI - A follow-up study of seasonal affective disorder. AB - BACKGROUND: The long-term course of seasonal affective disorder has not been well studied. METHOD: Using the Structured Clinical Interview for DSM-III-R, we interviewed 75% of a sample of 124 subjects diagnosed from five to eight years previously as fulfilling DSM-III-R criteria for recurrent major affective disorder, seasonal pattern. RESULTS: In the follow-up period, 38% of the sample continued to fulfil DSM-III-R criteria for seasonal illness; 28% had recurrent major depressive disorder, but no longer displayed a seasonal pattern; 18% were completely well with no further depression; 6% had subsyndromal symptoms; and 5%, although not meeting DSM-III-R criteria for seasonal illness, were still displaying constant periodicity. A short duration of index episode and a high frequency of illness predicted a continuing seasonal course of illness. CONCLUSION: Diagnostic criteria for seasonal affective disorder need to be further refined, possibly restrictively, if they are to be used to predict the future course of seasonal illness. PMID- 7496650 TI - Investigation by linkage analysis of the XY pseudoautosomal region in the genetic susceptibility to schizophrenia. AB - BACKGROUND: A susceptibility locus for schizophrenia in the pseudoautosomal region has been proposed on the basis of a possible excess of sex chromosome aneuploidies among patients with schizophrenia and an increased sex concordance in affected sib pairs. Several studies investigating this hypothesis have produced conflicting evidence. METHOD: In a series of Icelandic and British families, we used lod score and sib pair linkage analyses with markers for the MIC2 and DXYS14 loci on the pseudoautosomal XY region. RESULTS: Lod and sib pair linkage analysis with these markers produced strongly negative scores. Heterogeneity testing also produced negative results. CONCLUSION: We conclude that the present study provides no support for the involvement of either the pseudoautosomal region or the nearby region of the sex chromosomes in the aetiology of schizophrenia. PMID- 7496649 TI - Anorexia nervosa in 'non-white' populations. AB - BACKGROUND: Comparisons were made between two groups of anorectic patients, 'non white' and 'white', on a variety of clinical and social characteristics. METHOD: The study is based on a 34-year long database (1960-93) associated with a national tertiary referral centre. Thirty-six non-white patients meeting DSM-III R criteria for anorexia nervosa (AN) and five with 'partial syndrome' are compared with a white group comprising 944 patients with full syndrome. RESULTS: The main finding is clinical similarity between the two groups, extending to social class and pathological patterns of family relationship. The non-whites are shorter in stature (P = 0.004) and report earlier menarche (P = 0.004); they are younger at presentation (P < 0.001), somewhat less emaciated, and practice veganism slightly more commonly; they less often acknowledge sensitivity to 'fatness' (P < 0.003). This sensitivity was exposed later whenever treatment involved substantial weight gain. The proportion of non-white cases accepted for assessment has not changed in respect of year of onset (around 6%) for the last 20 years, or year of presentation (around 6-7%) over the last 15 years. There was a tendency for non-white patients to be referred earlier in their illness. CONCLUSIONS: The clinical and background profiles of non-white and white anorectics are generally similar. The numbers arising and presenting have not changed recently. Non-white patients apparently have as ready access as white patients to assessment and treatment by us. PMID- 7496651 TI - Neuropsychological slowness in obsessive-compulsive patients. Is it confined to tests involving the fronto-subcortical systems? AB - BACKGROUND: Although it is acknowledged that obsessive-compulsive (OC) patients may be slower than healthy controls in performing neuropsychological tests, speed has usually been treated as a confounding variable. It is possible, however, that the slower performance of OC patients is itself the result of a dysfunction of specific neural circuits (in particular of fronto-subcortical systems). METHOD: A neuropsychological battery including tests sensitive to fronto- and temporo subcortical dysfunction was administered to a group of OC patients and a group of healthy controls. Each test provided independent indices of accuracy and speed. RESULTS: OC patients were significantly slower than controls only when performing tasks involving the fronto-subcortical systems, whereas they did not differ from controls with respect to accuracy indices. CONCLUSION: It may be that neuropsychological slowness of OC patients is not merely an epiphenomenon of meticulous concern for correct test execution or intrusion of obsessive thoughts, but reflects the dysfunction of fronto-subcortical systems. PMID- 7496652 TI - Does rational role-play enhance the outcome of exposure therapy in dysmorphophobia? A case study. AB - BACKGROUND: Dysmorphophobia may improve after a variety or combination of treatments, but which elements of a treatment are useful is usually not certain. METHOD: In a single out-patient case study rational role-play ('paradoxical discourse') was added to exposure plus cognitive restructuring (CR) for a dysmorphic delusion study comprised of 20-minute sessions one week apart. RESULTS: Previous exposure plus attempted CR had improved anxiety, phobias, work and social leisure, but not dysmorphic belief. Added brief rational role-play was followed by resolution of the dysmorphic belief. All measures remained much improved at 18-month follow-up. CONCLUSIONS: Rational role-play deserves a controlled trial in dysmorphophobia and other problems and exploration of its mechanism of action. PMID- 7496653 TI - Russian literature and psychiatry. PMID- 7496654 TI - Psychological debriefing. PMID- 7496655 TI - Life events and dementia. PMID- 7496656 TI - Community care in presenile Alzheimer's disease. PMID- 7496657 TI - Repeat prescription antidepressants and residential care. PMID- 7496658 TI - Clozapine-induced hypotension treated with moclobemide and Bovril. PMID- 7496659 TI - Extreme suicidality following serotonin syndrome. PMID- 7496660 TI - Brain damage and tardive dyskinesia. PMID- 7496661 TI - Psychosurgery for obsessional disorder. PMID- 7496662 TI - Treatment of obsessive-compulsive disorder. PMID- 7496663 TI - Clozapine-induced hypersalivation and the alpha 2 adrenoceptor. PMID- 7496665 TI - Behavioural toxicities of antidepressants. PMID- 7496664 TI - Suspected congenital sertraline dependence. PMID- 7496666 TI - Laser cosmetic surgery. PMID- 7496667 TI - Laser skin resurfacing. Pre- and posttreatment guidelines. AB - BACKGROUND: Carbon dioxide laser skin resurfacing using either ultrapulsed lasers or scanning skin lasers has evolved as an effective method of treating photodamaged skin. OBJECTIVE: The purpose of this paper is to describe appropriate pre- and posttreatment regimens to ensure the optimum response to laser therapy. METHODS: We describe our experiences with 30 patients treated with laser skin resurfacing using different pre- and posttreatment regimens. Patients were evaluated by physicians as regards to their responses to and recovery from laser therapy. Some patients agreed to skin biopsy and cutaneous patch testing of topically applied agents. RESULTS: Eighty percent of patients achieved good to excellent improvement. Sixty-five percent of patients undergoing laser skin resurfacing developed contact dermatitis to several topical agents. Patch testing was negative on normal skin for contact allergy, suggesting a primary irritant reaction in laser-treated skin. Post-laser hyperpigmentation and erythema were the most noted immediate and mid-term side effects to laser resurfacing. An optimum pretreatment regime includes topical retinoids, skin lightening agents, and, immediately pretreatment, oral anti-herpes simplex medication plus oral antibiotics. An optimum posttreatment regimen includes minimizing topical therapy with the use of dilute acetic acid facial soaks. In addition, oral anti-herpes simplex medications and broad spectrum antibiotics are continued for 7 days post laser resurfacing. CONCLUSION: Laser skin resurfacing is an evolving means of treating photodamaged skin. Carefully selected pre- and posttreatment regimens, ideally under dermatologic supervision, are required to obtain optimum results. PMID- 7496668 TI - Laser skin resurfacing with the SilkTouch flashscanner for facial rhytides. AB - BACKGROUND: Effective treatment of facial rhytides has been reported using carbon dioxide (CO2) lasers with high peak power and short exposure time which creates char-free ablation. Char-free ablation can also be created using a Silktouch flashscanner attached to a conventional CO2 laser. OBJECTIVE: The purpose of this study is to evaluate the effectiveness of the SilkTouch flashscanner in skin resurfacing. METHODS: The SilkTouch flashscanner attached to one of two continuous wave CO2 lasers was used to treat facial rhytides on 40 patients. Histopathology to evaluate the depth of penetration of the scanner on both CO2 lasers was performed on preauricular skin prior to excision during facelift surgery. Silicone surface replicas were obtained pre- and 2 months post-laser treatment on two patients and evaluated by optical micrometry. Clinical evaluation of all patients pre- and post-laser treatment was performed. RESULTS: Clinical evaluation showed significant improvement of facial rhytides. Optical micrometry revealed a decrease in rhytide volume, indicating rhytide improvement. CONCLUSION: The Silktouch flashscanner is effective for the treatment of facial rhytides. PMID- 7496669 TI - Skin resurfacing with the Ultrapulse carbon dioxide laser. Observations on 100 patients. AB - BACKGROUND: The Ultrapulse carbon dioxide (CO2) laser has been shown to be a highly effective treatment for photodamaged skin including improvement of rhytides, precancerous and benign skin lesions, as well as superficial benign pigmented lesions. OBJECTIVE: We describe our experiences using the Ultapulse CO2 laser in treating 100 patients with different severities of photodamaged skin. METHODS: All patients had moderately or severely photodamaged skin and were selected for treatment with the Ultrapulse CO2 laser to evaluate the efficacy and toxicity of this treatment. The patients were placed on a pretreatment regimen for 2-6 weeks prior to receiving treatment with the Ultrapulse CO2 laser. Patients were evaluated for pre- and posttreatment severity of photodamage. In addition, skin surface replicas were obtained in selected patients and evaluated by computer image analysis technology. RESULTS: At 1 month post-laser treatment, 68 patients received a moderate improvement, five patients achieved a marked improvement, and the remaining 27 patients showed minimal improvement. By 2 months post-laser treatment, 20 of the 27 patients who at 1 month showed only minimal improvement revealed a moderate to marked improvement from baseline. All the patients studied developed a transient erythema that lasted up to 6 weeks, and many of these patients showed a transient hyperpigmentation that persisted for up to 4 months. CONCLUSION: The Ultrapulse CO2 laser is confirmed as an effective treatment for skin resurfacing of photodamaged skin. PMID- 7496670 TI - Rapid laser scanning for facial resurfacing. AB - BACKGROUND: Laser skin resurfacing is growing in popularity in both the professional as well as lay populations. Freehand delivery of impacts is relatively time consuming. Accurate and precise placement of impacts to avoid unwanted overlapping and the superimposing of multiple impacts is difficult. OBJECTIVE: To evaluate a new computerized scanning device, the computerized pattern generator (CPG), and to compare it with freehand delivery of multiple laser impacts over large areas. METHODS: Sixty-one patients were treated using the CPG by surgeons who are experienced with freehand laser resurfacing. RESULTS: CPG delivery provides more rapid resurfacing with more uniform, accurate, and precise distribution of impacts. CONCLUSION: Combining the CPG with the Ultrapulse laser is a valuable addition making the procedure faster and more controllable. PMID- 7496671 TI - Laser resurfacing in pigmented skin. AB - BACKGROUND: Laser skin resurfacing is a very promising new treatment for solar elastosis and acne scars. The Ultrapulse carbon dioxide (CO2) laser and the SilkTouch flashscanner allow precise depth control and char-free ablation. Earlier studies have been done mainly on Caucasian skin types I and II, but very little data were available on pigmented skin. OBJECTIVE: The purpose of this study is to assess the efficacy and safety of laser skin resurfacing in skin types III and IV. METHODS: Thirty Asian and Hispanic patients were treated for facial rhytides and acne scars. All patients were instructed to use tretinoin cream 0.05%, hydroquinone 5%, and desonide 0.1% cream nightly for 2-4 weeks prior to the laser treatment. The Ultrapulse 5000C CO2 laser with the Truespot 3-mm collimated handpiece at the setting of 250-450 mJ per pulse, or the Silk-Touch flashscanner at the setting of 5-7 W, 0.2-second pulse duration, and 4-mm (M) spot size, was used. Patients were evaluated clinically, and global response was assessed by both clinicians and patients. RESULTS: The facial rhytides and acne scars improved 25-50% in all patients after one laser treatment. The most common side effects were persistent erythema that resolved on the average of 6 weeks. Hyperpigmentation occurred, but was reduced with regular use of tretinoin, hydroquinone, and desonide cream both pre- and postoperatively along with use of broad spectrum sunscreen after treatments. CONCLUSION: Laser skin resurfacing can be used to treat facial rhytides and acne scars in skin phototypes III and IV. When proper pre- and postoperative management is implemented, the risk of dyspigmentation can be reduced. PMID- 7496672 TI - Laser-assisted rhytidectomy. A preliminary report. AB - BACKGROUND: The carbon dioxide (CO2) laser is a surgical tool recognized for its capacity of combined simultaneous tissue transection and hemostasis without unwanted distant transmission of energy. OBJECTIVE: To determine the potential of laser-assisted rhytidectomy. METHODS: A preliminary group of 18 patients were treated for facial rejuvenation by a technique of laser-assisted rhytidectomy with excellent results. RESULTS: No major complications including hematoma were seen. CONCLUSION: Based on these initial findings, we conclude that laser assisted rhytidectomy shows great promise for better results with diminished risks. PMID- 7496673 TI - Laser hair transplantation. Tissue effects of laser parameters. AB - BACKGROUND: Micro- and minigrafting have become increasingly popular in hair transplantation over the past decade, resulting in larger numbers of grafts per session and lengthy, potentially technically difficult procedures. The advent of very high energy pulsed carbon dioxide lasers as surgical instruments has led to the investigation of their use in hair transplantation, as bloodless surgery with tissue removal by vaporization has obvious advantages as far as surgical speed is concerned. METHODS: A prospective study of eight patients comparing conventional micro- and minigrafting using 18- and 14-gauge needle punctures with dilators versus laser-vaporized donor sites with variation of laser parameters was conducted with monthly evaluation of healing and hair growth for 6 months. In addition, an in vitro histological study of laser vaporization recipient sites on excised scalp tissue was conducted to evaluate the residual thermal damage of various laser parameters. RESULTS: The conventional micro- and minigrafts healed faster and resulted in better hair growth than the laser vaporization sites. The donor sites were judged to be comparable as far as healing, but more time consuming with the laser. The operative time for the laser recipient sites was 50% of that of the conventional sites. CONCLUSIONS: The clinical results of laser hair transplantation are very dependent upon the laser parameters and technique used. Though operative time may be significantly reduced, the end result clinically is dependent on proper choices of pulse parameters and technique. PMID- 7496674 TI - Laser hair transplantation III. Computer-assisted laser transplantating. PMID- 7496675 TI - Mini-slit graft hair transplantation using the Ultrapulse carbon dioxide laser handpiece. AB - BACKGROUND: The new Ultrapulse carbon dioxide (CO2) laser technology has added a new dimension to many cosmetic surgery procedures including hair transplantation. Early reports by Unger and David (Laser Hair Transplantation. J Dermatol Surg Oncol 1994;20:515-21) have been encouraging with the potential of minimal bleeding, ease of placing transplanted grafts, and an overall shortened operative time. A 2-mm slit handpiece has been recently created to expedite this procedure. OBJECTIVE: The purpose of this study is to further investigate the use and efficiency of the new Ultrapulse CO2 laser slit handpiece in hair transplants. METHOD: Mini-slit graft hair transplants using the new Ultrapulse CO2 laser slit handpiece were done on 25 patients in 30 transplant sessions. Donor minigrafts were obtained by the strip harvesting technique using a triple-blade scalpel. Approximately 200-400 recipient slits were made with the 2-mm slit handpiece at the laser setting of 350 mJ, 12 W, 0.8 seconds per pulse. RESULTS: All grafts were easily placed into recipient sites with minimal bleeding and charring. The procedure was done in half the time of the conventional non-laser technique. Postoperatively, patients were quite satisfied with little pain and swelling. Histologic exams of the laser-treated slits showed minimal adjacent tissue necrosis. Long-term follow-up visits showed good regrowth of hair in these grafts. CONCLUSION: The new Ultrapulse CO2 laser slit handpiece proved to be an effective tool for mini-slit graft hair transplantation. PMID- 7496676 TI - Current trends in laser blepharoplasty. Results of a survey. AB - BACKGROUND: The use of lasers in cosmetic surgery has been expanded fairly recently to include blepharoplasties. Controversy exists as to the efficacy of this procedure. This survey is designed to gain an understanding of the efficacy of blepharoplasty performed by laser versus conventional scalpel techniques. METHODS: A group of surgeons who perform the procedure were questioned. Over 4,000 cases of upper and lower lid laser blepharoplasty were reported by the responding physicians. RESULTS: The results of this survey indicate that carbon dioxide is by far the most common laser used in laser blepharoplasty (96%). A laser is used as a sole cutting tool by 70% of the responders and as a sole hemostasis tool by 88% of surgeons. Overall, both the intraoperative time and postoperative recovery period were reported as significantly reduced when blepharoplasty is performed by laser as compared with scalpel. In general, the incidences of edema, ecchymosis, and postoperative pain were reported as less severe with laser. No serious complications related to the laser were documented by the survey responders. CONCLUSIONS: This study confirms that laser blepharoplasty techniques can be safe and effective in skilled hands and may even have some potential advantages over conventional blepharoplasty. PMID- 7496677 TI - Carbon dioxide laser ptosis surgery combined with blepharoplasty. AB - BACKGROUND: Repair of levator aponeurosis pathology has become accepted technique for the correction of acquired ptosis. Extensive experience with carbon dioxide (CO2) laser blepharoplasty has been obtained. OBJECTIVE: CO2 laser blepharoplasty techniques are expanded to repair acquired ptosis. METHODS: A step-wise outline of laser surgical technique is presented. RESULTS: A retrospective serial study of 82 patients is reviewed and compared favorably with other reported series. Improvement in appearance and reduction of asymmetry to less than 1 mm was observed in 69 patients (84%). Thirteen patients had more than 1 mm of asymmetry and eight underwent surgical revisions. No long-term complications such as chronic exposure keratitis were noted. CONCLUSIONS: The CO2 laser repair of acquired ptosis by aponeurotic surgery can be combined with blepharoplasty surgery. Improved intraoperative hemostasis, decreased operating time, and improved appearance in the immediate postoperative period are achieved with this technique. PMID- 7496678 TI - Lateral canthal tendon suspension using the carbon dioxide laser. A modified technique. AB - BACKGROUND: The "tarsal strip" procedure was developed to correct lower eyelid ectropion caused by lateral canthal tendon laxity. OBJECTIVE: The "tarsal strip" procedure is improved by using the carbon dioxide (CO2) laser. METHODS: The CO2 laser was used to perform lateral canthotomy. Patients were evaluated postoperatively for subjective improvement in preoperative symptoms. Complications such as epithelial inclusion cysts were actively sought. RESULTS: Thirty-one patients (60 lids) were followed for an average of 12 months. Preoperative symptoms resolved in 86% (N = 27), improved in 7% (N = 2), and did not change in 7% (N = 2). Two patients required additional surgery. Complications were not observed in this study. CONCLUSION: The modified "tarsal strip" procedure using the CO2 laser provides many advantages compared with the original procedure. A clean dissection improves visibility and allows easy identification of the lateral tarsal plate. The lateral canthal angle is preserved and epithelial inclusions are prevented because the conjunctival mucosa is not violated. PMID- 7496680 TI - Estimating the volume of lung irradiated during tangential breast irradiation using the central lung distance. AB - The aim of this study was to evaluate the central lung distance (CLD) as a surrogate for the volume of lung irradiated during tangential breast radiotherapy. 20 women underwent a planning computed tomography (CT) scan and three-dimensional (3D) treatment planning for both breasts. The field size was perturbed in a systematic manner to give a number of plans with the CLD increasing from 0 to 30 mm. The volume of lung irradiated was determined directly using dose-volume histograms of the lung and correlated with the known CLD. The results indicate that absolute and percentage lung volumes increase with increasing CLD according to a quadratic relationship which is different for both left and right lungs. It is concluded that while there is no substitute for a 3D plan and a lung dose-volume histogram calculation, CLD may be used as a guide to the volume of lung included within the tangential fields used for breast radiotherapy. PMID- 7496679 TI - An experimental investigation of uroselectan B. 1932. PMID- 7496681 TI - Quality control of low dose craniospinal irradiation for low risk medulloblastoma. AB - Between July 1984 and May 1990, 25 children (median age 9 years) classified as low risk medulloblastoma were treated post-operatively with low dose craniospinal irradiation (25 Gy) followed by a boost to the posterior fossa (total dose 55 Gy), with or without chemotherapy. Simulator and portal films were reviewed for all patients. Doses delivered at the mid cerebral and spinal axis, anterior vertebral body, field junctions and posterior fossa were recalculated and correlated with outcome. Three children were excluded because of inadequate data leaving 22 cases for study. The 5-year actuarial survival is 55%. Nine recurrences and one second tumour (astrocytoma) were observed. Sites of recurrence were either in the posterior fossa (5), in the brain (3), in the spinal axis (1), or combined (2). The dose delivered was within +/- 5% of that prescribed to the brain in 21/22 cases, and to the posterior fossa and the anterior surface of the vertebral body in 17/22 cases. The only patient who received less than 95% of the prescribed dose to the brain (23 Gy) failed in the subfrontal area and olfactory plate. One of the five patients who received less than 95% of the prescribed dose to the posterior fossa (50.3 Gy) failed at the primary site. None of the five patients who received less than 95% of the dose to the spinal axis failed. Quality control of radiation treatment showed that failures could not be correlated with incorrect technique. PMID- 7496682 TI - Technical note: femoral angiography using the reverse shuffle technique. AB - We report our findings using the "reverse shuffle" technique of stepping table femoral angiography. This technique represents a simple modification of the conventional stepping table sequence entailing a reverse table movement allowing acquisition of early and late films in the distal positions. It was developed in order to minimize the number of runs required in each examination. In a retrospective study 52/53 (98%) of angiograms using the reverse shuffle technique required only a single run compared with 30/51 (58%) using the conventional technique. We believe that the reverse shuffle should be used where possible when stepping table angiography is performed. Furthermore, the incorporation of a reverse table movement may also be advantageous in angiography using automated digital subtraction angiography (DSA) stepping systems. PMID- 7496683 TI - Technical note: inspiration or expiration? Reducing motion artefact in digital subtraction arch angiography of the extracranial carotid arteries. AB - 100 consecutive patients underwent digital subtraction aortic arch angiography for suspected carotid bifurcation disease. Patients were prospectively randomized to suspend respiration in inspiration or expiration during image acquisition. Images were assessed qualitatively as "excellent", "adequate" or "poor". 92 studies were available for review. Adequate and poor groups were combined into a non-excellent group due to the small numbers that were scored poor. There were 27 (53%) excellent, 24 (47%) non-excellent in the expiration group and 12 (29%) excellent, 29 (71%) non-excellent in the inspiration group. Analysis of angiogram scores gave chi 2(1) = 4.3; p = 0.038, indicating that the higher angiogram scores of the expiration group were statistically significant. Pixel shifting was used in 61 cases (66.3%). The proportion was 34.4% lower in the expiration group (95% CI; 17-52%) with chi 2(1) = 10.5; p = 0.0012 indicating that the reduction in the use of pixel shifting for the expiration group was statistically significant. The superiority of arch angiograms obtained with the patient suspending respiration in expiration rather than inspiration has been demonstrated. This was achieved with significantly reduced use of pixel shifting, encouraging greater confidence in the validity of the angiographic findings. The technique is free of cost or time implications. PMID- 7496685 TI - Case report: malignant mesothelioma of the tunica vaginalis testis--an indolent course. AB - Malignant mesothelioma of the tunica vaginalis testis is an extremely rare tumour. So far less than 50 cases have been reported in the literature. Radical inguinal orchiectomy appears to be the optimal treatment. Upon recurrence, treatment remains unsatisfactory. A further case is reported here, where the disease ran an indolent course despite several recurrences after initial treatment. PMID- 7496684 TI - Technical note: the relationship between TV position and the effectiveness and comfort of protective spectacles in fluoroscopic procedures. AB - Although interventional radiological procedures can result in significant eye doses to an operator, protective measures are often rejected because screens are found to impede manipulations, and 0.5 mm lead equivalent spectacles are heavy and uncomfortable. With many units the TV monitors are positioned so that the operator needs to turn his head through 80 degrees or more to view the image while screening. The aim of this study was to determine the distribution of the shielding in protective spectacles which will combine maximum dose reduction for different TV positions with minimum weight. Thermoluminescent dosimeters were used to assess the effectiveness of various spectacles in shielding the lens of the eye when a skull phantom was orientated at different angles in a large area beam of X-rays generated at a tube potential typical of diagnostic energies. The beam was tilted upwards at an angle of 20 degrees as though originating from the irradiated volume of a patient. The results showed that if weight was a consideration, and the head was turned away from the image intensifier through an angle greater than 80 degrees, the shielding was most effective when concentrated in the side-panels of the spectacles. The shape of the side-panels, however, was critical to the adequate shielding of this radiation. PMID- 7496686 TI - Case report: non-malignant cause of brachiocephalic vein compression following treatment for lung cancer. AB - Obstruction of venous return in the mediastinum or thoracic inlet is well recognized, particularly in oncological practice. In most cases the obstruction is of the superior vena cava. For such venous obstruction to occur following treatment for lung cancer usually implies malignant lymph node involvement and therefore a grave prognosis. This report describes a patient who developed unilateral signs of impaired venous return from the head, upper trunk and upper limb following treatment for lung cancer. The cause was found to be compression of the left brachiocephalic vein by normal anatomical structures as a result of mediastinal shift following pulmonary lobectomy and radiotherapy. This case, together with others reported in the medical literature, suggests that the development of isolated brachiocephalic vein obstruction does not necessarily have the same ominous implications as the development of superior vena caval obstruction. PMID- 7496687 TI - Case report: haemangiopericytoma of the oesophagus. AB - Haemangiopericytomas are rare, vascular soft tissue sarcomas typically located in the retroperitoneum, pelvis or lower extremities. To our knowledge this neoplasm within the oesophagus has not been reported previously. We present a patient with a haemangiopericytoma of the oesophagus and discuss the radiographic and pathological findings. PMID- 7496688 TI - Case report: coincident granular cell tumour of the breast with invasive ductal carcinoma. AB - Granular cell tumours of the breast are rare lesions which are almost always benign. They are of interest because their clinical, radiological and pathological appearances often closely resemble those of malignancy. A case is described in which screening mammography in an asymptomatic woman led to the diagnosis of simultaneous invasive intraductal carcinoma in one breast and benign granular cell tumour in the other. Granular cell tumour of the breast coincident with other breast tumours is very rarely described and an association with breast carcinoma is unknown. PMID- 7496689 TI - Case of the month: differential calculus. PMID- 7496690 TI - Factors in dependent lobar super-enhancement during CT arterial portography. PMID- 7496691 TI - Sixth nerve palsy following radiculography. PMID- 7496692 TI - Invited review: computer aids for decision-making in diagnostic radiology--a literature review. AB - This review looks at a variety of different ways in which computers can be used to assist in the interpretation of radiological images and in radiological decision-making. The issues involved in the design of computerized decision aids are introduced and four criteria proposed for evaluating such aids: need, practicality, veracity and relevance. These criteria are used to assess research into decision aids based on: image databases, numerical methods, expert systems, image processing and image understanding systems. Possible directions for research leading to aids of practical value are discussed in the conclusion. PMID- 7496694 TI - Body magnetic resonance angiography and its effect on the use of alternative imaging--experience in 1026 patients. AB - Time-of-flight magnetic resonance (MR) angiography has been described for numerous applications but the number of patients described in published reports is usually relatively small. There is little information concerning the general utility of MR angiography in the body, outside the cranial circulation, or its effect on use of other imaging studies. Utilization of two-dimensional (2D) time of-flight MR angiography over a 3 year period was reviewed to determine its accuracy and impact on other imaging modalities. Cranial and cardiac studies were excluded. Correlation was made with alternative imaging and surgical findings. Between January 1990 and December 1992 2D time-of-flight MR angiography was used to examine 1026 patients. MR angiography was used most frequently to examine the venous system. There was a slight reduction in the use of alternative imaging with two exceptions. When chest MR venography was performed, alternative imaging was completely abolished. Also, contrast portal venography was virtually eliminated. Five errors (0.2% of total MR angiography examinations) were recorded. Time-of-flight MR angiography, especially MR venography, is accurate and can replace contrast venography for abdominal and thoracic applications. Cost and limited availability means that other imaging, usually ultrasound, often precedes MR angiography. PMID- 7496693 TI - Comparison of single reading with double reading of mammograms, and change in effectiveness with experience. AB - In the prevalence round of screening for breast cancer at a single centre, the benefit of the second radiological report has been assessed using data from 33,734 women, their screening results and knowledge of the interval cancers. The service was set up under the UK National Breast Screening Programme, and the data show evidence of learning by both individuals and the team as a whole. Of the 269 cancers detected by screening, 33 would not have been diagnosed if the only report available had been the first. The recall recommendation rate associated with the first report was 6.9% and 236 cancers were detected. The recall recommendation rate associated with all queries of all the reports was 10% (3354 queries). Had these 3354 queries all been investigated (instead of the 1423 actually recalled) only a further three cancers would have been detected. Sensitivity of the programme as a whole was substantially better than that of individual radiologists, while the specificity was maintained. The decision pathway by which recalls were agreed between the two radiologists resulted in a low recall rate (4.2%) for the programme as a whole, and is a critical factor in gaining the benefit of the improved sensitivity without a concomitant deterioration in the specificity. With the passage through the prevalent round, recall rates steadily fell, the malignant to benign biopsy ratio improved and sensitivity increased. The second radiological report yielded 14% additional cancers diagnosed and contributed very significantly to good sensitivity and so to the effectiveness of screening. Economic analysis of the results will be reported in a further paper. PMID- 7496695 TI - Metastatic breast carcinoma involving the gastric antrum and duodenum: computed tomography appearances. AB - We describe the computed tomography (CT) appearances of three cases of antroduodenal linitis plastica metastases from breast carcinoma. Two of the three cases had biliary obstruction as a consequence and required endoscopic stenting. Antroduodenal linitis plastica should be considered as a possible cause for jaundice in patients with breast carcinoma. PMID- 7496696 TI - A comparison of iodixanol with iopamidol in aorto-femoral angiography. AB - This double-blind, randomized, parallel group clinical investigation in 140 consecutive patients undergoing aorto-femoral arteriography was carried out to compare iodixanol (Visipaque) 270 mgI ml-1 with iopamidol (Iopamiro) 300 mgI ml 1. The aims of the study were to compare adverse events and discomfort, clinical chemistry parameters in blood, haemodynamics and diagnostic information of the angiograms in the two groups. The main parameter for statistical analysis was the visual analogue scale (VAS) score for overall discomfort experienced by the patients during the examination. 134 patients, 69 and 65 receiving iodixanol and iopamidol, respectively, were examined according to the protocol and included in the evaluation. The two groups of patients were judged to be comparative. Statistically significant milder discomfort was felt with iodixanol than with iopamidol (p = 0.0001); mean VAS values 16 mm and 51 mm, respectively. Pain was reported far less frequently after iodixanol than after iopamidol (7.4% versus 50.8%) whereas sensation of warmth was less intense after iodixanol than after iopamidol. Four patients in the iodixanol group and two in the iopamidol group reported transient, non-serious adverse events. The difference was not statistically significant (p = 0.68). Systolic blood pressure was affected to a slightly greater degree after injection of iopamidol than after injection of iodixanol. Measurements of diastolic blood pressure, as well as clinical chemistry parameters in blood, revealed no changes of clinical importance, and all arteriograms performed were of diagnostic value. The conclusion is that iodixanol 270 mgI ml-1 is as efficacious as iopamidol 300 mgI ml-1, but produces less discomfort during arteriography. As such, iodixanol is a good alternative to iopamidol in aorto-femoral angiography. PMID- 7496697 TI - Magnetic resonance characteristics of intrapelvic haematomas. AB - Although haematomas in the female pelvis are not rare, their magnetic resonance (MR) appearance has not been well characterized. Accordingly, we analysed the MR appearances of various stages of haematomas in the female pelvis. A retrospective analysis of 35 haematomas in 28 patients sequentially imaged with a 1.5 T MR unit was performed. The time interval between the insult and MR imaging was determined and the appearances of the haematoma were evaluated. The results were compared with those in the central nervous system. The evolution of haematomas in the pelvis appeared similar to that of haematomas in the brain, and four stages (acute, early subacute, late subacute and chronic) were identified according to the signal intensity pattern on T1 and T2 weighted images. Pelvic haematomas differed from intracranial haematomas in the following features: (1) The speed of evolving pelvic haematomas was gradual. (2) In the centre of acute haematomas, slightly hyperintense areas were seen on both sequences, suggesting residual oxyhaemoglobin. (3) Haematomas evaluated in the early subacute stage had complex appearances. (4) In older haematomas, central intermediate signal intensity areas, presumably due to diamagnetic hemichromes, became predominant on T1 weighted images. In conclusion, although the evolution of a pelvic haematoma is similar to that of a brain haematoma, its speed is gradual and MR appearances are somewhat different due to environmental differences. PMID- 7496698 TI - High resolution ultrasound anatomy of normal Achilles tendon. AB - To evaluate the ultrasound (US) appearance of the normal Achilles tendon at increasing frequency and establish an anatomical correlation for US findings, 30 normal tendons were examined in vivo and three in vitro with 10 MHz and 15 MHz mechanical sector probes. Side-by-side comparison was performed in vitro between the sonograms and the corresponding anatomical sections. Two tendinous portions were detected by presence of an internal acoustic interface which had different appearances: one (type I) or two (type II) continuous lines of increased thickness and greater reflectivity than adjacent fibrils; or displacement (type III) of the distal portion of the well insonated sector of the tendon body. When, on coronal scans of the tendon, no intratendinous linear echoes of increased reflectivity were visible, the two portions of the tendon were identified through the converging course of their bundles (type 0 pattern). Different echogenicity allowed the detection of two tendinous portions, also on axial images. Scanning of isolated tendons allowed precise location of these interfaces at the boundary between anatomically distinct tendinous portions arising from the soleus and gastrocnemius muscles. Although the normal Achilles tendon is commonly regarded as a uniform structure by US, the use of high resolution probes allows identification of its constituent portions. Their identification may be useful to avoid misdiagnoses of pathological findings. PMID- 7496699 TI - The impact of digital imaging on patient doses during barium studies. AB - Barium studies performed on 10 digital and four non-digital fluoroscopic systems were monitored with dose-area product meters as part of a Regional Patient Dosimetry Audit programme. The data have been collected using a computer to read and reset the dose-area product meter and also to collect patient and examination details. A comparison of dose-area product measurements from digital and non digital fluoroscopy units on over 10,000 barium studies is presented. The data have been corrected according to patient size. The mean size corrected dose-area product for a barium meal examination was found to be 7.62 Gy cm2 for a digital set compared with 15.45 Gy cm2 for a non-digital set with 2462 and 1308 patients included in each measurement series, respectively. Dose-area products were also a factor of approximately two lower for barium enema, barium swallow and barium follow-through examinations performed on digital systems. PMID- 7496700 TI - In vivo induction of apoptosis in human lymphocytes by therapeutic fractionated total body irradiation. AB - Ionizing radiations have been reported as an in vitro apoptosis initiating stimulus in human lymphocytes. As the cytotoxicity of ionizing radiations and chemotherapeutic agents appears to be dependent on the efficacy of cell death induction, the manipulation of apoptosis initiation might be used as a means to supress some pathological process. In the present study the in vivo induction of gamma-ray mediated programmed cell death in humans is reported. The in vivo induction of apoptosis in peripheral blood lymphocytes (PBL) by ionizing radiations was investigated in 33 patients after each of two sessions (2 Gy and 4 Gy) of fractionated total body irradiation (FTBI) as part of their conditioning regimen before bone marrow transplantation. PBL committed to apoptosis were scored before irradiation (S1), 4 h (S2) and 24 h after 2 Gy (S3, 14-17 h after the second 2 Gy fraction). Nuclear morphology and chromatin-DNA were analysed by fluorescence microscopy immediately after blood sample withdrawal (I) and after 24 h in cell culture medium (II). When scored immediately after withdrawal, no circulating PBL with the apoptotic nuclear morphology were observed in S1 and S2 blood samples whereas S3 disclosed 21.9 +/- 11.7% of circulating lymphocytes with an apoptotic nuclear morphology. After 24 h in culture, S1 samples (before irradiation) generally contained less than 20% of apoptotic lymphocytes. A higher percentage of apoptotic cells was noted in some cases in relation with recent chemotherapy and possibly with pathology. After 24 h in culture, S2 and S3 samples contained 51.7 +/- 17.9% and 60.4 +/- 16.4% of apoptotic lymphocytes, respectively. These results confirm that ionizing radiations induce apoptosis in vivo in human lymphocytes and that the commitment to apoptosis can be determined after low doses (2 Gy) of therapeutic whole body irradiation. The results suggest that susceptibility to apoptosis induction by ionizing radiations could be related to previous therapy by cytotoxic drugs and possibly to the type of haematological malignancy. PMID- 7496701 TI - The application of automatic computing machines to radiation treatment planning. 1954. AB - A new method of determining the dose distribution required in treatment planning has been developed by using punched cards, and sorting and tabulating machines instead of isodose charts. The percentage depth doses produced by an X-ray field are first recorded on several sets of punched cards. The number of sets required for one field depends upon the distances used from the cross point of the multifield axes to the point of entry. Each set consists of 36 cards, with each card recording the percentage depth doses of 15 points in one radial line, taking the cross points of central axes of the fields as the origin. In all cases the points are taken at the distances 0, 1, 2, 3, 4, 5, 6, 8, 10, 12, 14, 16, 18, 20, 24 cm from the origin and along radial lines 10 deg. apart. A special polar co ordinate dose distribution sheet has been designed, and the contour of the patient's body and the position of the fields designated for treatment are drawn on this sheet. Instead of putting the isodose charts at the proper positions of the fields, the sets of punched cards for the fields to be applied are automatically arranged in the right order. Instead of the time-consuming process of reading off the percentage depth doses from the overlapping isodose charts and adding them up for selected points, the cards are fed into the automatic tabulating machine, which makes the summation of data for all the points mentioned above and tabulates the results. The whole operation is done in 10 to 15 minutes. The tabulated results are then plotted on the special dose distribution sheet. A generalised mathematical treatment of the dose at any point in the irradiated region is discussed, and an equation of "point-tumour dose ratio" is derived. Further application of this equation is made to special cases for treatment using equal maximum doses, equal tumour doses and rotation therapy. The geometrical principles involved are also indicated. PMID- 7496702 TI - Errors at knee magnetic resonance imaging: true or false? AB - Amongst a series of 332 patients undergoing a prospective evaluation of the influence of magnetic resonance imaging (MRI) on diagnosis and outcome, arthroscopic correlation became available in 92 knees (91 patients). The MRI report, the arthroscopic findings and the patient's case notes were reviewed by a consensus panel. With respect to the menisci and cruciate ligaments, the formal radiological report was at variance with the arthroscopic findings in 22/349 sites where there had been no previous surgery (10 medial meniscus, six lateral meniscus and six anterior cruciate ligament). 12 of these 22 errors were considered, on review, to be genuine MRI errors. However, considerable controversy remains as to whether the other 10 "errors" were true or false. There are several factors which contribute to such controversy. These include technically difficult arthroscopies, delays between MRI and arthroscopy, and ambiguities in the wording of both the referral letter and the radiological report. These factors should be considered when evaluating the diagnostic performance of both MRI and arthroscopy. PMID- 7496703 TI - Evaluation of palpable breast masses with 201Tl scintigraphy. AB - 39 female patients (age range: 31-84 years) with palpable breast masses detected by physical examination, underwent 201Tl scintigraphy in order to assess its value in the detection of breast carcinomas and to differentiate them from benign breast masses. Planar images were carried out at 20-30 min and 2-3 h after intravenous administration of 111-185 MBq (3-5 mCi) of 201Tl chloride. In 12 patients single photon emission tomography (SPET) studies were also performed. In 18 patients the scintigraphic studies were positive and in 17 of these cases, breast carcinomas were confirmed. Tumour sizes ranged from 1.3 to 6 cm in diameter. In the remaining patient a false positive result was obtained where there was benign breast change. In three of seven cases, malignant axillary nodes were also detected. All 21 patients with negative scintigraphy had benign breast lesions. There were no differences between images obtained at 20-30 min and 2-3 h or between planar images and SPET studies. In 10 patients there was disagreement between mammography and 201Tl scans. 201Tl scans confirmed the presence of carcinoma in three cases and discarded malignancy in the other six cases. In the remaining case, 201Tl scan was false positive. 201Tl scintigraphy is useful in distinguishing malignant from benign breast masses, even when compared with mammography. PMID- 7496704 TI - Computed tomography manifestations of tricuspid regurgitation. AB - Six patients undergoing dynamic contrast enhanced abdominal computed tomography (CT) for upper abdominal symptoms and signs are described. Each patient demonstrated early, intense opacification of the inferior vena cava (IVC) and hepatic veins, exceeding that in the aorta on scans through the upper abdomen. Opacification of the IVC returned to normal on more caudal slices. All six patients were found to have echocardiographically proven tricuspid regurgitation (TR). It is suggested that this sign occurs in patients with TR due to direct reflux of contrast from the right atrium into the IVC during protracted bolus injection. Additional CT findings included patchy parenchymal liver enhancement in four patients and ascites in five. Recognition of this simple sign may direct the clinician to a cardiac aetiology for upper abdominal symptoms and signs. PMID- 7496706 TI - A linear modelling approach to automatic interpretation of quality control measurements in mammography. AB - An approach to the automated interpretation of quality control data is described in which the relationship between small changes in system performance and the resulting small changes in the measured quality control test results are approximated as a set of linear equations in matrix form. The inverse of this forward matrix is then used to identify causal changes from the set of measured changes. This approach has been investigated for the case of mammographic screening quality assurance and shown to provide a potentially useful tool to assist in the interpretation task. PMID- 7496705 TI - Validation of 99Tcm-HMPAO leucocyte scintigraphy in ulcerative colitis by comparison with histology. AB - Leucocyte scintigraphy offers an alternative to more invasive techniques in the investigation of inflammatory bowel disease. The accuracy of 99Tcm-HMPAO leucocyte scintigraphy has not been assessed by comparison with colonic histology, which was the aim of this study. 15 patients with ulcerative colitis underwent 99Tcm-HMPAO leucocyte scintigraphy (TLS) less than 5 days before colonoscopy. Histological features of mucosal biopsies were compared with total and segmental colonic TLS scores. Segmental and total scintigraphy scores correlated most strongly with histological grades for acute inflammation (r = 0.75, p < 0.001 and r = 0.9, p < 0.001, respectively) and chronic inflammatory cell infiltration in the lamina propria (r = 0.76, p < 0.001 and r = 0.86, p < 0.001, respectively). 99Tcm-HMPAO leucocyte scintigraphy detected acute inflammation in the colon of patients with ulcerative colitis with a sensitivity of 91% and negative predictive value of 80% and localized acute inflammation to a particular colonic segment with a sensitivity of 82%, specificity of 94%, accuracy of 88%, positive predictive value of 94% and negative predictive value of 91%. 99Tcm-HMPAO leucocyte scintigraphy positivity predicts and localizes colonic acute inflammation with a high degree of confidence, but negative scintigraphy does not exclude acute inflammation. PMID- 7496707 TI - An investigation into techniques for reducing doses from neo-natal radiographic examinations. AB - A survey of radiation doses received by infants having radiography in a neo-natal unit was undertaken at Aberdeen Maternity Hospital. Options for dose reduction recommended by the CEC Lake Starnberg Group were investigated. Techniques were implemented for clinical evaluation after assessments of image quality with test objects and phantoms. Clinical image quality of radiographs was evaluated by paediatric radiologists using CEC criteria. Results revealed a clear relationship between entrance dose and patient weight, indicating that reference doses for infants could be linked to weight. At the start of the study the mean entrance dose for chest anteroposterior (AP) radiographs was 65 microGy and the dose-area product 12.3 mGy cm2 for a system with 2.5 mm aluminum filtration using 50 kVp X rays with a 200 speed class film/screen system. A 400 speed class film/screen combination was investigated but not evaluated clinically, because the image quality was not considered acceptable. Addition of 0.1 mm copper to the filtration of the X-ray tube reduced entrance doses by 50%, but the technique was not adopted, because of a poorer image quality with only 72% of features in clinical images being reproduced to an acceptable standard, compared with 87% with the original system. Use of faster processing chemicals, an increase in filtration to 3.5 mm aluminium and a tube potential of 60 kVp resulted in a 40% reduction in mean entrance dose to 37 microGy with satisfactory clinical image quality. PMID- 7496708 TI - A phantom based method for deriving typical patient doses from measurements of dose-area product on populations of patients. AB - One of the chief sources of uncertainty in the comparison of patient dosimetry data is the influence of patient size on dose. Dose has been shown to relate closely to the equivalent diameter of the patient. This concept has been used to derive a prospective, phantom based method for determining size correction factors for measurements of dose-area product. The derivation of the size correction factor has been demonstrated mathematically, and the appropriate factor determined for a number of different X-ray sets. The use of phantom measurements enables the effect of patient size to be isolated from other factors influencing patient dose. The derived factors agree well with those determined retrospectively from patient dose survey data. Size correction factors have been applied to the results of a large scale patient dose survey, and this approach has been compared with the method of selecting patients according to their weight. For large samples of data, mean dose-area product values are independent of the analysis method used. The chief advantage of using size correction factors is that it allows all patient data to be included in a survey, whereas patient selection has been shown to exclude approximately half of all patients. Reduction of the size of the data set may lead to mean dose-area product values that are less reliable indicators of typical practice. The use of size correction factors will be of particular benefit in the analysis of paediatric dosimetry data, where a wide range of sizes exist, even within accepted age bands. PMID- 7496709 TI - How reliable are ultrasound measurements of renal length in adults? AB - Ultrasound assessment of patients with renal impairment commonly includes measurement of bipolar renal length. Reduction in length is considered to indicate chronic renal disease and is a factor in deciding whether to proceed to renal biopsy. To date, no published data are available on interobserver and intraobserver variation in sonographic renal length measurement in adults. Bilateral renal lengths were measured in 20 adult subjects, with no history of renal disease, by three experienced operators, on two separate occasions. Limits of agreement for replicate measurements by each ultrasonographer and for replicate measurements by each pair of ultrasonographers were determined. Values of repeatability (a measure of intraobserver variation) and reproducibility (a measure of interobserver variation) were calculated for all renal length measurements, and for right and left renal lengths separately. Results indicate that replicate renal length measurements differ by less than 1.85 cm in 95% of cases, and the magnitude of variations is similar when measurements are made by either single or different ultrasonographers, and are similar for right and left renal length measurements. This suggests that sonographic bipolar renal length measurements in normal adult kidneys are reasonably reliable. In diseased kidneys, however, in which identification of renal poles is difficult, interobserver and intraobserver variation may be much greater. PMID- 7496710 TI - An assessment of the intrauterine sound intensity level during obstetric echo planar magnetic resonance imaging. AB - In order to assess the sound level experienced by the fetal ear during obstetric magnetic resonance imaging, a fluid filled stomach was used as an experimental model of the gravid uterus. A better than 30 dB attenuation in intensity was recorded across the frequency band of interest for all patient orientations. This was enough to reduce acoustic sound pressure from a level close to the instantaneous damage threshold (120 dB), to an acceptable level (< 90 dB). Direct mechanical coupling through the patient table was also shown to increase uterine sound pressure levels by as much as 10 dB. Much higher peak pressures could be obtained by tapping of abdomen with the fingers. PMID- 7496711 TI - Technical note: magnetic resonance imaging-guided pre-operative breast localization using "freehand technique". AB - A 1.5 cm focus in the breast with intense gadolinium uptake was incidentally detected during magnetic resonance (MR) imaging evaluation of a cluster of microcalcifications. Because the apparent mass was both mammographically and sonographically occult, MR imaging was used to guide a needle by "freehand technique" to the lesion, permitting successful pre-operative localization and subsequent surgical excision of the area. PMID- 7496713 TI - Technical note: comparison of colour Doppler energy sonography with conventional colour Doppler sonography in detection of flow signal in peripheral renal transplant vessels. AB - The technical advantage of colour Doppler energy sonography (CDE) results in a higher sensitivity for blood flow detection than in conventional colour Doppler sonography (CDI). We compared the efficiency of CDE versus CDI in the detection of blood flow signals in asymptomatic patients after renal transplantation. 10 asymptomatic voluntary patients after renal transplantation were evaluated with CDI and subsequently with CDE by two examiners. Filter, scale and gate setting were kept constant, the Doppler gain was increased individually in every patient until background noise just did not derange the image. The assessment of blood flow signal was done using a self defined score system. CDI showed Doppler signal in the interlobar vessels in all 10 patients and blood flow signal in arcuate arteries and adjacent portions of interlobular vessels in nine of 10 patients. We were not able to detect blood flow related signal in the more peripheral medullary and cortical vessels. CDE obtained blood flow signal in interlobar and interlobular vessels in all patients. In addition, Doppler signal was observed in all patients in small medullary and cortical vessels. CDE increases detection of blood flow related signal in peripherally located small medullary and cortical vessels. There was no improvement in the evaluation of blood flow in interlobar vessels but marked improvement in blood flow detection in small peripherally located vessels. CDE cannot completely replace CDI in the sonographic evaluation of vascular complications in renal transplants, but should be used as an additional measure to rule out pathology in small peripheral renal vessels. PMID- 7496712 TI - Technical note: dynamic analysis of the pharynx during swallowing using Turbo FLASH magnetic resonance imaging combined with an oral positive contrast agent--a preliminary study. AB - Serial images of the swallowing motions of 10 healthy volunteers were taken using the Turbo-fast low angle shot (FLASH) magnetic resonance (MR) imaging sequence, while the subject swallowed an oral contrast agent containing ferric ammonium citrate. In all cases, the flow of the contrast agent and the motion of the organs in the oral cavity and the pharynx during swallowing could be observed. High-speed MR imaging combined with an oral positive contrast agent has the potential to evaluate accurately swallowing motion. PMID- 7496714 TI - Technical note: an evaluation of a Diamentor based system for estimation of spot film dose-area product values. AB - In the audit of barium studies it is not common to assess the contribution of spot films to the total dose. This study investigates the accuracy with which the number of spot films and associated dose-area product (DAP) may be estimated. The study was undertaken for barium enemas, the swallow part of barium swallow studies, and the abdominal part of barium meal studies on X-ray sets with film screen radiography, digital fluorography and 100 mm camera fluorography. DAP readings from a Diamentor were input to a computer at a rate of three per second and the difference in successive DAP values calculated. These data were used to assess the number and total DAP from spot films. For Diamentor based systems which sample the DAP values at three times per second the threshold algorithm works well for film-screen radiography of barium meals and enemas. However, the estimated number of spot films is generally in error. For the Diamentor M2 it is unlikely that the sample rate can be increased to a high enough value to provide sufficient accuracy for all barium studies obtained using all spot film modalities, although use of the modified threshold algorithm has the potential to provide some increase in accuracy. PMID- 7496715 TI - Technical note: potentially higher patient radiation doses using digital equipment for barium studies. AB - Digital fluorography offers several advantages over conventional radiographic systems. The general expectation of digital equipment has been of a lower radiation dose to the patient. A series of dose-area product measurements on two digital sets during barium meal and barium enema examinations has shown that this expectation is not necessarily correct, particularly when the dose from the fluoroscopic part of the examination is considered. Users of digital fluorography sets are advised to check the performance of their equipment by suitable dose measurements. PMID- 7496716 TI - Technical note: non-surgical treatment of epiphora by balloon dacryocystoplasty- the technique. AB - Dacryocystoplasty (DCP) was offered as an alternative to patients waiting for dacryocystorhinostomy (DCR) for epiphora. 31 procedures were undertaken with a technical success rate of 93% and a clinical improvement in 89% of these patients. It is concluded that DCP should be the technique of choice in the initial management of epiphora due to stenosis or occlusions of the nasolacrimal ducts before contemplating a DCR. PMID- 7496717 TI - Case report: malignant fibrous histiocytoma following radiation therapy of fibrous dysplasia. AB - Malignant fibrous histiocytoma commonly occurs spontaneously. In some cases it follows previous therapeutic or incidental irradiation, or miscellaneous pre existing osseous conditions. Recently, it has been associated with total hip arthroplasty. We report a case of malignant fibrous histiocytoma following radiation therapy of fibrous dysplasia and review the relevant literature. PMID- 7496718 TI - Case report: late aggressive meningioma following prophylactic cranial irradiation for acute lymphoblastic leukaemia. AB - The clinical, radiological and pathological findings in a 28-year-old female patient who developed aggressive meningioma 20 years after prophylatic cranial irradiation (PCI) for acute lymphoblastic leukaemia (ALL) are described here. Only four cases of late atypical/aggressive meningioma following PCI were detected in a thorough search of the literature. The high cure rate in childhood ALL, attributable to aggressive chemotherapy and PCI, is capable of inducing secondary brain tumour, including aggressive meningioma. PMID- 7496719 TI - Case report: chronic mesenteric ischaemia as a cause of abnormal bowel uptake of labelled leucocytes. AB - Chronic mesenteric ischaemia is often difficult to diagnose. A high level of clinical suspicion is needed and further investigation requires invasive procedures. We present a case where weight loss and abdominal pain were dominant features and initial interpretation of a labelled leucocyte scan suggested inflammatory bowel disease. Subsequent investigations excluded this possibility and chronic ischaemia was confirmed at autopsy. Positive small bowel images using 99Tcm-HMPAO labelled leucocytes may indicate chronic ischaemia. PMID- 7496720 TI - Case report: fibromatosis of the breast in a male patient. AB - Fibromatosis of the breast is an uncommon disease, although several series of this condition in females have been reported in the literature. We present the first case to be described in a male patient. PMID- 7496721 TI - Case of the month: a painful red eye. PMID- 7496722 TI - Short communication: breast parenchymal patterns and their changes with age. AB - In studies on mass screening, it has often been reported that tumours in breasts with dense parenchyma are difficult to detect and may have a more advanced stage at diagnosis. Shorter rescreening intervals have been suggested for these women but, before recommending such a strategy, it is important to investigate how often dense breast parenchyma (P2 and DY patterns according to Wolfe) is actually present in a screening population and to what extent these patterns change with age. The prevalence of dense breast parenchyma (P2 and DY) in our study population was 33% at first examination (n = 2581), which is fairly low compared with other screening populations. Its presence was strongly, inversely age dependent. Breast patterns of 1177 women, aged 35-85 years, were followed for 12 years. In 39% (182/461) of the women with a P2 or DY pattern at their first examination, regression to a lucent pattern (N1 and P1) occurred over the years. The majority of these women were assumed to have reached menopause in the follow up period. These findings support the hypothesis that the presence of dense breast parenchyma is related to the reproductive period and indicate that shortening the rescreening intervals would be most effective in pre-menopausal age groups. PMID- 7496723 TI - Radiation dose to the lens from CT of petrous bones. PMID- 7496724 TI - Comparison of light reflection rheography with ascending venography in the diagnosis of lower limb venous thrombosis. PMID- 7496725 TI - Receptor mediated glycotargeting. AB - Glycotargeting relies on carrier molecules possessing carbohydrates that are recognized and internalized by cell surface mammalian lectins. Numerous types of glycotargeting vehicles have been designed based on the covalent attachment of saccharides to proteins, polymers and other aglycones. These carriers have found their major applications in antiviral therapy, immunoactivation, enzyme replacement therapy and gene therapy. This review compared different types of glycotargeting agents and the lectins which have been successfully targeted to treat both model and human diseases. It may be concluded that the discovery of new mammalian lectins which endocytose their ligands will lead to the rapid development of new glycotargeting agents founded on the principles of carbohydrate-protein interactions. PMID- 7496726 TI - Ocular permeability of FITC-dextran with absorption promoter for ocular delivery of peptide drug. AB - The purpose of this study is to characterize an ocular permeability of FITC dextran, as a model of peptide drug, and to evaluate the effects of absorption promoters on the ocular permeability of FITC-dextran. The in vitro penetrations of FITC-dextrans (average molecular weight 4400 and 9400: FD-4 and FD-10) were measured across the isolated corneal and conjunctival membranes of albino rabbits using a two-chamber glass diffusion cell. The corneal permeabilities of FD-4 and FD-10 were much lower than the conjunctival permeabilities. Scraping of corneal epithelium extremely increased the corneal permeabilities. The penetration parameters were estimated according to Fick's equation. Absorption promoters such as EDTA, taurocholic acid, benzalkonium chloride and saponin significantly increased corneal permeabilities of FD-4 and FD-10. Saponin showed the highest promoting activity. Conjunctival permeabilities of FD-4 and FD-10 were also enhanced by absorption promoters although the improvements of conjunctival permeabilities by absorption promoters were smaller than those of corneal permeabilities. Ratios of corneal to conjunctival permeabilities were enhanced by absorption promoters. These results indicate that an ocular delivery of instilled hydrophilic macromolecule is markedly low and a selective use of absorption promoter can improve the extent and pathway of its ocular absorption. PMID- 7496727 TI - Modulation of murine macrophage nitric oxide synthesis by liposomal phospholipids: correlation with liposome immune adjuvant activity. AB - The influence of alum and liposomal phospholipids on interferon-gamma-(IFN gamma), IFN-gamma/N-acetylmuramyl-L-alanyl-D-isoglutamine- (MDP) or IFN gamma/tumor necrosis factor-alpha- (IFN-gamma/TNF-alpha) induced macrophage nitric oxide (NO) synthesis has been investigated. IFN-gamma induced NO synthesis in a dose-dependent manner. TNF-alpha and MDP did not induce NO synthesis, but interacted synergistically with sub-optimal doses of IFN-gamma. Alum strongly inhibited IFN-gamma-induced NO synthesis (ID50 25 microgram/ml). Liposomes composed of dipalmitoylphosphatidylcholine (DPPC) had no effect on IFN-gamma induced NO synthesis. IFN-gamma-induced NO synthesis was stimulated by DPPC/dimyristoylphosphatidylglycerol (DMPG) liposomes (9:1 mol ratio, ED50 45 nmol phospholipid/ml), and inhibited by DPPC/dipalmitoylphosphatidylethanolamine (DPPE) liposomes (9:1 mol ratio, ID50 > 500 nmol phospholipid/ml), and DPPC/phosphatidylserine (PS) liposomes (7:3 mol ratio, ID50 150 nmol phospholipid/ml). Alum, DPPC/PE and DPPC/PS liposomes also inhibited IFN gamma/MDP- and IFN-gamma/TNF-alpha-induced NO synthesis. Neither alum or the liposome preparations had significant toxicity towards macrophages in vitro at concentrations that induced maximal inhibition or stimulation of IFN-gamma induced NO synthesis. Immunization of mice with alum-adsorbed and liposome incorporated bovine serum albumin (BSA) demonstrated that enhancement or reduction of both IgG antibody and the proportion of IgG2a/IgG2b was correlated with stimulation or inhibition of IFN-gamma-induced NO synthesis. PMID- 7496728 TI - In vivo disposition characteristics of plasmid DNA complexed with cationic liposomes. AB - To control disposition and hence gene expression, we investigated the disposition characteristics of plasmid DNA complexed with the cationic liposomes Lipofectin and LipofectACE after intravenous injection in mice via the tail vein. The optimum ratios of DNA and liposome complexes were selected through in vitro cytotoxicity and transfection studies. The highest transfection was found at the DNA:liposome ratio of 1:5 w/w. Hence, this ratio was used for in vivo disposition studies, and the distribution patterns were compared with that of naked pCAT. Following intravenous injection of [32P] pCAT, radioactivity was rapidly eliminated from plasma and approximately 60% of the dose was taken up by the liver within 1.5 min. In the case of LipofectACE samples, radioactivity elimination from plasma was equally rapid, but its accumulation was observed in both the liver (35%) and the lung (45%). For Lipofectin samples, radioactivity was initially accumulated in both the liver (55%) and the lung (25%), but lung accumulation was not sustained beyond 5 min after administration. Both liposomal samples showed in vivo gene expression in the lung, heart, kidney and spleen, but not in the liver. Thus, the present study demonstrated that disposition and gene expression of pCAT can be controlled by complex formation with liposomes. PMID- 7496729 TI - Pharmacokinetics of [3H]biotin bound to different avidin analogues. AB - The use of avidin-biotin technology in drug delivery facilitates the conjugation of biotinylated therapeutics to transport vectors that are enabled to undergo receptor-mediated transcytosis through the brain capillary endothelial wall, which makes up the blood-brain barrier (BBB) in vivo. However, the conjugation of avidin, a cationic glycosylated protein, to transport vectors greatly increases the rate of removal of the vector from the bloodstream, owing to rapid uptake of avidin by peripheral tissues such as liver and kidney. However, modified avidins may retain high affinity biotin binding properties, but may not be rapidly removed from plasma by peripheral tissues, and such avidin analogues would provide preferred plasma pharmacokinetic profiles. Therefore, the present studies investigate the pharmacokinetics of plasma removal of [3H]biotin bound to one of six different avidin analogues: streptavidin, Neutra-lite avidin, avidin, neutral avidin, Lite-avidin, and succinylated avidin. Isoelectric focusing studies show that avidin and Lite-avidin were highly cationic proteins, whereas neutral avidin, Neutra-lite avidin, and streptavidin were neutral proteins, and succinylated avidin had an acidic isoelectric point. The avidin analogues fell into two groups with respect to rate of biotin removal from plasma. The low clearance group included streptavidin and Neutra-lite avidin, which had a mean plasma clearance of 0.41 mL/min/kg. The high clearance group consisted of succinylated avidin, neutral avidin, and Lite-avidin and had a mean plasma clearance of 17 mL/min/kg, or 40-fold faster than the low clearance avidins.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7496730 TI - A new method to isolate polyalkylcyanoacrylate nanoparticle preparations. AB - A simple method for the separation of polyalkylcyanoacrylate nanoparticles was developed using polyisohexylcyanoacrylate (PIHCA) as a model. Fluorescein isothiocyanate dextran 70 was used to label the nanoparticles. Ultracentrifugation onto a performed sucrose gradient allowed the easy elimination of the dextran and of the free molecules remaining in the upper phase. After such treatment, the physicochemical characteristics of the PIHCA nanoparticles were not modified. This method could be usefully extended to other types of nanoparticles. PMID- 7496731 TI - Preparation and evaluation of a time-controlled release capsule made of ethylcellulose for colon delivery of drugs. AB - A novel ethylcellulose (EC) capsule which releases drug with a time-controlled fashion has been prepared. This capsule is composed of four parts, drug container, swellable substance, capsule body and cap. At the bottom of the body, micropores are made. As water penetrates through these micropores, the swellable substance such as low substituted hydroxypropyl cellulose (L-HPC) swells. When the cap made of water-insoluble macromolecular substance such as EC cannot persist the swelling pressure, the EC cap disintegrates and the drug in the container is released from the capsule. The lag-time is utilized for the delivery of drug to the colon. The release time of the drug from the capsule was measured both in vitro and in vivo experiments. In the case of an in vitro experiment, after 12mg of fluorescein as a model drug and 238mg of starch were filled into the container, caps having different thickness were attached to the capsule body and release study was performed. The release time of the drug was mainly dependent on the thickness of the cap. Using test capsules of which mean cap thickness were 39.1 +/- 2.3 (SE)microns, 63.1 +/- 5.0 microns and 75.6 +/- 4.1 microns, the in vivo release time was estimated after administration to beagle dogs. As a parameter, the peak time (tmax) when plasma fluorescein concentration reached to its maximum level was determined for the estimation of the release time of the drug from the capsule in the gastrointestinal tract. The in vivo tmax was well correlated with the cap thickness.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7496732 TI - Delivery of protein antigen to the major histocompatibility complex class I restricted antigen presentation pathway. AB - Major histocompatibility complex (MHC) class I-restricted antigen presentation normally requires a protein antigen to be synthesized in the cytosol of the antigen presenting cell (APC). Exogenous protein antigen could gain access to the class I presentation pathway if the protein is introduced into the cytosolic compartment of the APC. Approaches which release the protein antigen from endocytic vesicles have been employed to deliver protein antigen for the recognition by class I-restricted cytotoxic T lymphocytes (CTL). These include osmotic shock, electroporation, cationic and pH-sensitive liposomes. An alternative approach is to deliver a gene that encodes the protein antigen. In this case, the APC is transfected with a gene which synthesizes the "exogenous protein" in the cytosol. Delivery of protein antigen targeted for CTL induction in vivo follows a different strategy and generally requires an antigen carrier of lipidic/membranous nature, such as liposomes, immunostimulating complexes, and/or lipid conjugates. Macrophages that are responsible for scavenging the antigen play an important role in CTL induction. An optimal CTL inductive vaccine must contain other immuno-modulatory activities in addition to its activity in delivering antigen to the class I pathway. Attempts to attenuate viral infection and to improve anti-tumor immunity have been successful by delivering the exogenous antigen entrapped in liposomes. These animal model studies should be of great value in the development of potential vaccine formulation. PMID- 7496733 TI - PCNA defined proliferative activity of epithelial tumor cells in adenomas of the colon. AB - Proliferative activity of epithelial tumor cells was evaluated with the use of immunohistochemistry and anti-PCNA monoclonal antibodies in alcohol fixed paraffin embedded sections of 44 colonic adenomas, including 33 tubular, 5 villous and 6 tubulovillous adenomas. The mean PCNA index was 24.7 +/- 10.9%, 24.8 +/- 6.2% and 24.8 +/- 14.0% in tubular, villous and tubulovillous adenomas respectively. In 12 tubular adenomas with dysplasia the mean PCNA index in areas with dysplasia was significantly higher (38.2 +/- 11.5%) as compared to areas without dysplasia (17.0 +/- 8.9%; p < 0.05). The results indicate that PCNA index of epithelial tumor cells is significantly increased in adenomas with high grade of dysplasia irrespective of histological type or size of the tumour. PMID- 7496734 TI - PCNA and protein P53 in recurrent supratentorial glial brain tumors: studies on correlation between morphology and tumor progression. AB - Thirty seven cases of recurrent gliomas with survival time from 3 month to 13 years were investigated morphologically and immunohistochemically to evaluate progression and transformation of tumors. Using antibodies against PCNA and p53 the proportion of positively marked cells (nuclei) to all tumor cells was assessed at both primary and secondary surgery. This was compared with morphology and survival time between first and second surgery. In 10 cases (27.0%) there were no positively marked cells. In the remaining 27 cases (73.0% of the whole group), the following indices (percents of positive cells) were calculated: for patients with survival below 1 year (mean = 9m) 28.3% PCNA-positive cells and 30.3% p53-positive cells at first operation. For recidiving tumors both indices were lower namely for PCNA 20.9% and for protein p53-26.7%. For patients with survival over 1 year (mean = 4.8y) the index after I operation for PCNA was 25.5% and was higher than found after II operation-22.3%. In contrary, the index for protein p53 was 20.1% after I operation and was lower as compared with the index after II operation-28.2%. Eleven of twelve fibrillary astrocytomas and all five gemistocytic astrocytomas at first operation underwent changes towards malignisation. The immunocytochemical results confirm the high phenotypic differentiation what is reflected in different indices for PCNA and p53 of tumors. PMID- 7496736 TI - Comparative distribution of S-100 protein and antigen HMB-45 in various types of melanomas and naevi. AB - The content of S-100 protein and antigen HMB-45 in various types of melanomas and skin naevi was compared with respect to the frequency of two antigens, their distribution within lesions as well as diagnostic or prognostic relevance. The study material comprised 72 skin melanomas, 25 melanomas of the oral cavity and 63 non-malignant skin naevi. Formalin fixed, paraffin-embedded sections were used for immunohistochemical reactions with polyclonal rabbit anti-S-100 antibody (Dako) and monoclonal anti-HMB-45 antibody (Dianova) followed by PAP and APAAP Kits (Dianova), respectively. Detection of HMB-45 was preceded by a microwave treatment of sections. S-100 protein was found in all cases of melanomas irrespective of their location or histological type, but HMB-45 was missing in 8% of oral melanomas. Distribution of the antigens within tumors was heterogeneous and often mutually exclusive. S-100 protein was also present in all types of naevi, while HMB-45 was absent in intradermal naevi and strictly confined to epidermal component in the remaining ones. The intensity of staining for S-100 correlated inversely with the patients' survival. It was concluded that demonstration of HMB-45 might facilitate a differentiation of naevi from melanomas, whereas staining for S-100 protein might have some prognostic significance. PMID- 7496735 TI - Abnormalities of chromosome 5q correlate with morphologic features of better prognosis in clear cell renal carcinomas. AB - Aberrations of the long arm of chromosome 5 (5q) were studied in 79 clear cell renal carcinomas (clear cell RCC) by LOH (loss of heterozygosity) method, using microsatellite markers D5S107, CRTL1, LNS-CA, IL-9.RP1, CFS1R and GeneScan analysis software. Alterations of chromosome 5q were detected in 42% of cases. We found, that accumulation of genomic abnormalities at multiple loci of 5q, especially in 5q21-qter region, where genes for cytokines are located, correlates with morphologic features of better prognosis. PMID- 7496737 TI - Idiopathic membranous glomerulonephritis: a quantitative study of glomerular and interstitial lesions. AB - Twenty renal biopsy specimens from patients with idiopathic diffuse membranous glomerulonephritis (MGN), for whom both light and electron microscopy as well as immunofluorescence microscopy and full clinical data were available, were examined quantitatively and compared with fifteen cases of normal controls. Morphometric investigations were performed by means of a computer image analysis system to evaluate whether morphometric analysis may be helpful in morphological diagnosis of MGN, and to study whether serum creatinine and changes in quantitatively analysed glomeruli could correlate with the interstitial fibrosis. The study revealed that total glomerular area, total number of glomerular cells per total glomerular area, and total glomerular cells per unit of glomerular area as well as relative interstitial volume were significantly increased in MGN in comparison with normal glomeruli. Moreover, there was significant positive correlation between the width of the cortical interstitium and serum creatinine level but not between the width of the cortical interstitium and glomerular changes. These findings suggest, that in MGN mainly interstitial lesions are involved in the renal prognosis and they are not related to the severity of glomerular changes. The authors conclude, that although morphometric methods supported by image analysis system cannot be used instead of light microscopy, electron microscopy and immunohistochemistry, they may be helpful in every-day diagnostic procedures of early stages of MGN. PMID- 7496739 TI - Effect of misoprostol in ethanol-induced liver injury. AB - Prostaglandins appeared protective against acute experimental liver injury of different origin. Misoprostol, stable, orally active, synthetic derivative of PGE1 attenuates several functional alterations in liver mitochondria during ethanol administration. To study its possible hepatoprotective effects on ethanol induced liver injury in rats we measured: serum activities of alanine aminotransferase (ALT), gamma-glutamyltranspeptidase (GGT) and concentrations of ammonia in blood and liver tissue. Histopathological evaluation of liver slices was also performed. Activities of both enzymes and ammonia values were elevated after intragastric ethanol administration for 60 days. Treatment for 30 days with misoprostol resulted in their decrease. This effect was not observed in the control group. Beneficial results were also obtained in histopathological evaluation of the liver tissue. These results indicate potential therapeutic effects of misoprostol on ethanol-induced liver injury in rats. PMID- 7496740 TI - Clinical and morphological analysis of endometriosis in women of the Lublin Macroregion between 1987 and 1992. AB - Histopathological examinations performed between 1987 and 1992 at the Department of Pathology of the Lublin Medical Academy were analysed statistically. Endometriosis was found in 885 cases, i.e. 13.3% of all gynecological cases. An increased frequency as well as a shift in the peak age of incidence to the third decade, were found. PMID- 7496738 TI - Comparison of morphological and biochemical changes of BAL-isolated cells in experimental lung emphysema. AB - The aim of the study was to evaluate the protease and antiprotease activity in the fluid obtained from the culture of cells isolated from the lungs of animals with experimental emphysema. An attempt was made to correlate the results of biochemical examinations with adherence degree and ultrastructural changes of the surface of BAL-isolated cells. The experiment was carried out on male Wistar rats, of 180-220 g b.w. Two i.p. injections of BCG-vaccine (4 x 10(8) microorganisms) on the 1st and 14th day were applied as macrophage mobilizing and activating agent. Papain (2 mg/l ml/100 g b.w.) was given once i.t. on the 21st day. The animals were sacrificed on the 28th day of the experiment. We found a correlation between the increase in the cell adherence and ultrastructural changes (in SEM), suggesting an increased activity of the cells isolated from BCG treated rats. In the culture medium of cells isolated from the rats which were given BCG or papain and BCG+papain we observed an increased base protease activity and decreased Cathepsin D activity comparing with the control group. Increased antitrypsin activity in the BCG and BCG+papain-treated rats and decreased antitrypsin activity in papain-treated rats only was observed, too. There was no obvious difference in the levels of the antiplasmin and antichymotrypsin activities between the groups. The present results indicate that activated pulmonary macrophages are one of the sources of the protease antiprotease intraalveolar imbalance. However, an increased production of proteolytic enzymes may not be the only factor responsible for the progression of lung emphysema in BCG-treated rats.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7496741 TI - Glomerulonephritis associated with malignant diseases of non-renal origin. A report of three cases and a review of the literature. AB - Glomerular lesions have been reported to occur in association with a wide variety of malignancies, particularly carcinomas and lymphomas. Patients with the neoplastic diseases are exposed to continuous antigenemia, which stimulates antibody production and forms circulating immune complexes. Membranous nephropathy appears to be the most common glomerular lesion in patients with solid tumors, and minimal change glomerulopathy is another major form of glomerular disease associated with lymphomas, particularly with Hodgkin's disease. Three cases of nephropathy associated with anaplastic oat cell bronchial carcinoma, gastric adenocarcinoma and Hodgkin's disease were studied histologically, ultrastructurally and by immunofluorescence. The diagnosis of glomerulopathy preceded the diagnosis of malignant disease. All three patients were admitted to hospital because of nephrotic syndrome. The paraneoplastic glomerulopathies were histologically identical to that of idiopathic membranous glomerulonephritis and minimal change glomerulonephritis. Adult patients over the age of 50-60 with nephrotic syndrome caused by glomerulonephritis, especially by membranous nephropathy, should be screened for cancer. PMID- 7496743 TI - [SNOMed--international system for coding changes in medicine]. PMID- 7496742 TI - Usefulness of cytogenetic analysis in the diagnosis of undifferentiated synovial sarcoma of the chest wall. A case report. AB - A case of poorly differentiated spindle cell malignant neoplasm of the chest wall in a 21-year-old man is reported. The results of histopathological and immunohistochemical examinations were not characteristic enough to establish the precise diagnosis. Due to the age of the patient and the location of the lesion and NSE positivity of tumor cells, the possibility of Askin's tumour or malignant peripheral nerve sheath tumour was considered. Cytogenetic analysis revealed a chromosomal translocation t(X;18)(p11;q11), which is known as the specific aberration in synovial sarcoma. Chromosome study seems to be a useful tool for identifying undifferentiated mesenchymal tumours. PMID- 7496744 TI - Secondary brain insults after head injury: clinical perspectives. AB - Secondary brain insults, particularly hypotension (systolic blood pressure < 95 mm Hg) and hypoxia (PaO2 < 60 torr), are the most powerful determinants of outcome from severe head injury (SHI) that are amenable to therapeutic manipulation. Well over one quarter of SHI patients appear to suffer one or more secondary insults during the time between injury and resuscitation. Furthermore, although improved strategies for airway management have decreased the incidence of preresuscitation hypoxia, little progress has been made in ameliorating the influence of hypotension during this period. The incidence of hypotension in the ICU is also > 25%, and such episodes also appear to be significant predictors of poor outcome independent of their etiologies and of preresuscitation secondary insults. In all, over one third of all SHI patients appear to experience one or more secondary insults during their acute post-injury course, and these events are correlated with a doubling of mortality and a large increase in morbidity. The approach to proper management of such secondary brain insults requires only an increased recognition of their importance, greater vigilance toward monitoring, and improved application of therapeutic maneuvers that are generally already available. Given their prevalence and impact on outcome, prevention/correction of secondary brain insults may be the most powerful means of improving outcome from severe brain injury available today. PMID- 7496745 TI - Enhanced vulnerability to secondary ischemic insults after experimental traumatic brain injury. AB - Both experimental traumatic brain injury and clinical traumatic brain injury appear to render the brain more vulnerable to a second ischemic insult. The mechanisms of enhanced vulnerability to subsequent ischemia appear to include a reduced ability to increase cerebral blood flow in response to hypotension, hypoxemia, or acute anemia and increased tissue sensitivity to ischemia. Although numerous mediators may be involved in increased tissue sensitivity, those that particularly merit investigation include oxygen free radicals, glutamate, arachidonate metabolites, calcium ions, and protein kinase C. PMID- 7496746 TI - Cerebral blood flow in severe clinical head injury. AB - Following severe head injury, derangements of the cerebral vasculature and cerebral blood flow (CBF) often occur, rendering the brain at risk of secondary ischemia. Therefore, monitoring of CBF in head-injured patients is considered useful for understanding the pathophysiology and effects of therapy, although such monitoring has not yet become part of routine patient management in most centers. In this article, we review the current research on CBF in head injury. Understanding of the physiologic relationship between CBF and cerebral oxygen metabolism (CMRO2) is crucial in the interpretation of CBF values obtained in comatose head injured-patients. Although CMRO2 is reduced with coma, there is ample evidence to suggest that vulnerability of the brain to ischemia is in fact enhanced after traumatic injury. It is now well established that cerebral ischemia (CBF < or = 18 mL/100 g/min) is present in approximately 30% of cases within the first 6 hrs postinjury. In addition, early ischemia has been found to correlate with poor outcome and early mortality. Notably, early ischemia was present even with normal or restored blood pressure and arterial oxygenation, which suggests that other, nonsystemic causes of cerebral ischemia after traumatic brain injury exist. Although spasm of the larger cerebral arteries has been postulated as a possible cause of ischemia, recent measurements of cerebral blood volume are more compatible with compromise of the microcirculation, possibly due to perivascular swelling, with endothelial injury and leukocyte stasis. Disturbances of cerebrovascular CO2 reactivity and autoregulation appear to be less frequent than previously assumed. However, when present, such derangements do have consequences for therapy, in particular the management of blood pressure and cerebral perfusion pressure. Potential implications for patient management are discussed. PMID- 7496747 TI - Intracranial pressure monitoring techniques. AB - In patients requiring ICP monitoring, a ventricular catheter connected to an external strain gauge transducer or catheter-tip pressure transducer device is the most accurate and reliable method of monitoring ICP, and enables therapeutic CSF drainage. Clinically significant infections or hemorrhage associated with ICP devices causing patient morbidity are rare and should not deter the decision to monitor ICP. Parenchymal catheter-tip pressure transducer devices measure ICP, similar to ventricular ICP pressure, but have the potential for significant measurement differences and drift due to the inability to recalibrate. These devices are advantageous when ventricular ICP is not obtained or if there is obstruction in the fluid coupling. Subarachnoid or subdural fluid-coupled devices and epidural ICP devices are currently less accurate. PMID- 7496748 TI - Intracranial pressure and cerebral perfusion pressure in severe head injury. AB - Monitoring and management of intracranial pressure (ICP) are fundamental to modern neurotraumatology. Although never formally proven to independently improve outcome in prospective, randomized, placebo-controlled trials, there is such a predominance of indirect support for this modality that most neurotrauma protocols are impossible with-out its inclusion and ethical considerations virtually preclude placebo-controlled trials of its efficacy. In addition to the question of improving outcome, ICP monitoring is also useful in guiding the use of potentially harmful treatment modalities such as hyperventilation, mannitol, and barbiturates, and also provides important prognostic data. ICP monitoring provides information on the likelihood of cerebral herniation and allows calculation of the cerebral perfusion pressure (CPP). Although there is no constant threshold for herniation, the most commonly used treatment threshold is 20 to 25 mm Hg. In addition, ICP trends are indispensable in providing the earliest possible indication of critical intracranial mass effects when combined with other clinical indicators. CPP is the difference between mean arterial pressure and ICP. CPP is an important clinical indicator of cerebral blood flow (CBF). Cerebral autoregulation generally remains at least partially preserved after severe head injury, although the CPP value at which it is activated appears to be shifted upward. Therefore, maintaining adequate CBF appears to require using an elevated minimal CPP threshold when treating the injured brain. A generally accepted value of 70 mm Hg is suggested. PMID- 7496749 TI - Cerebral metabolic management. AB - Knowledge of cerebral metabolic variables increases the information available for managing the head-injured patient. This article reviews normal cerebral metabolism and describes the derangement of metabolism observed in patients with head trauma. Cerebral metabolism is globally decreased by one third to one half in the severely head-injured patient, usually because of the lower metabolic expenditure associated with coma, but sometimes because of superimposed hypoxia/ischemia, primarily due to secondary insults. Measurement of jugular venous oxygen saturation has become the most useful method for monitoring cerebral metabolism in the ICU. Although jugular venous oxygen saturation does not provide quantitative information about either cerebral blood flow or cerebral metabolism, it reflects the ratio between these two variables. To maintain a sufficient level of available oxygen and energy production at the cellular level, there are two therapeutic strategies: a) hemodynamic and b) metabolic. Treatments can be directed at both increasing oxygen delivery and integrating hemodynamic handling, and at various pharmacologic or physical methods intended to reduce the cerebral metabolic demand. The latter strategy is designed to depress either the basal or activation components of cerebral metabolism. This strategy includes manipulations of brain temperature and the use of central nervous system depressant, barbiturate, and similar drugs. PMID- 7496751 TI - Electrophysiologic monitoring in head injury. AB - Electrophysiologic monitoring of head-injured patients, using electroencephalogram (EEG) and sensory-evoked potentials, can provide a functional assessment of the neural structures involved in tested pathways. Experience using EEG, somatosensory-evoked potentials, brainstem auditory-evoked potentials, visual-evoked potentials, and the combination of sensory-evoked potentials termed multimodality-evoked potentials as clinical tools to augment patient evaluation and predict outcome demonstrates that these methods can be effectively integrated into clinical care. They can be used to augment the neurologic examination and provide useful information when other measures of neural function are limited by deep coma or by other treatment modalities that confound the clinical examination. PMID- 7496750 TI - Transcranial Doppler measurements. AB - Transcranial Doppler ultrasonography (TCD) has served a useful purpose in further defining the pathophysiology of disordered cerebrovascular control after head injury. The evaluation of altered cerebrovascular responses in individual patients may be useful in their clinical management. TCD can be a clinically useful tool in the early diagnosis of a variety of complications that can occur in head-injured patients, such as vasospasm, markedly increased intracranial pressure (ICP), low cerebral perfusion pressure, carotid dissection, and disordered cerebral autoregulation. Although the TCD waveforms alone can alert clinicians to the presence of markedly increased intracranial pressure, these waveforms are not specific for predicting moderate ICP increases. Experienced interpreters can identify TCD waveforms that indicate the arrest of the cerebral circulation. Cerebral circulatory arrest can be a useful confirmatory finding in the diagnosis of brain death in questionable cases, and also can be used to shorten the observation period in those patients in whom organ donation is contemplated or discontinuation of ventilatory support and nursing care may be indicated. PMID- 7496752 TI - Hyperventilation therapy for severe traumatic brain injury. AB - The management of brain swelling that frequently occurs following severe traumatic brain injury (TBI) presents a difficult challenge for physicians treating these patients. A traditional cornerstone for the treatment of post traumatic brain swelling has been prophylactic hyperventilation to reach PaCO2 levels of 25 to 28 torr. While there are anecdotal reports of improvement in intracranial pressure (ICP) and neurologic functioning following institution of this therapy, the only prospective, randomized trial of its use has found worse outcomes in those treated with prophylactic hyperventilation therapy for 5 days. That hyperventilation therapy might exacerbate secondary brain injury seems likely based on abnormalities in cerebral blood flow (CBF) and metabolism which result from TBI, and the potential for hyperventilation to worsen those abnormalities. Both global and regional CBF are critically reduced, and metabolism increased, during the first several hours and days after injury. As a result, focal ischemia is common following severe TBI. Hyperventilation causes a further decrease in CBF, often without a concomitant reduction in ICP. In some cases, TBI also causes an increase in cerebral vascular responsivity to hypocapnia, increasing the drop in regional CBF that occurs with hyperventilation. Thus, there is a well defined physiologic basis for expecting hyperventilation to cause worsened clinical outcomes following TBI. While this therapy clearly is indicated for the management of acute neurologic deterioration or intracranial hypertension refractory to all other forms of medical therapy, hyperventilation is no longer recommended as a first-line therapy for intracranial hypertension or as prophylactic therapy following severe TBI. PMID- 7496753 TI - Mannitol and other diuretics in severe neurotrauma. AB - Mannitol has replaced other diuretics as the agent of first choice for control of raised intracranial pressure (ICP) after brain injury. Mannitol should be given as a bolus intravenous infusion, over 10 to 30 mins, in doses ranging from 0.25 to 1.0 g/kg body weight. It may be given when high ICP is suspected, prior to computed tomography scanning, e.g., in patients who develop a fixed, dilated pupil or neurologic deterioration. This agent may also be used pre- or intraoperatively in patients with intracranial hematomas, and when high ICP is demonstrated in the ICU. It is more effective and safer when administered in bolus doses than as a continuous infusion. Mannitol may be safely used during the early resuscitation phase in hypovolemic patients with concomitant head injury, provided that plasma expanders and/or crystalloid solutions are given to correct the hypovolemia simultaneously. A Foley catheter should always be inserted when mannitol is used. Serum osmolality should be measured frequently after mannitol and maintained < 320 mOsm to avoid renal failure. Its beneficial effects and the rationale for its use are also reviewed. PMID- 7496754 TI - Steroids in head injury. AB - Failure of high-dose corticosteroid therapy to ameliorate intracranial hypertension or improve long-term neurologic outcome in patients with traumatic brain injury has been shown in several prospective, randomized clinical trials. Additionally, the risk of complications, including elevations in serum glucose levels and gastrointestinal hemorrhage, although relatively low, make routine use of glucocorticoids further unwarranted in head-injured patients. PMID- 7496755 TI - Sedative and neuromuscular blocking drug use in critically ill patients with head injuries. AB - Sedative and neuromuscular blocking (NMB) drugs are used to facilitate care of head trauma patients requiring mechanical ventilation or therapy of intracranial hypertension. Because no specific regimen is appropriate in all patients, drug selection and utilization exhibit significant regional variation. Sedatives are used to decrease anxiety and diminish awareness of noxious stimuli. Propofol offers particular promise in neurosurgical intensive care. NMB drugs are used in 1% to 10% or more of critically ill patients. Increasingly more information is available to guide the use of NMB drugs for patients suffering head trauma. Broad concerns about these drugs include their use as adjunctive therapy to control intracranial hypertension, the incidence of prolonged weakness or myopathy, the potential for direct neurologic toxicity, and their effect on outcome. Resolution of these issues will improve the use of sedative and NMB drugs in intensive care. PMID- 7496756 TI - High-dose barbiturates for intracranial pressure control. AB - Approximately 10% of severely head-injured patients will manifest elevated intracranial pressure uncontrollable by any currently known medical or surgical treatments, with a resulting mortality of > 90%. High-dose barbiturate therapy used in this setting has been shown to control intracranial pressure and reduce mortality in a small but definite number of patients. The pharmacologic, physiologic, and systemic effects of high-dose barbiturates must be fully appreciated, and appropriate clinical safeguards must be established when this therapy is implemented after severe head injury. PMID- 7496757 TI - Hypothermia and hyperbaric oxygen as treatment modalities for severe head injury. AB - Moderate systemic hypothermia has been shown to improve neurologic outcomes in both fluid-percussion and cortical contusion models of experimental brain injury. Based upon initial clinical work, it was concluded that at temperatures < 32 degrees C, patients with severe brain injury were at increased risk of ventricular arrhythmias, and that rapid rewarming immediately postinjury predisposed to intracranial pressure increases. Subsequent clinical studies of moderate hypothermia (32 degrees C) for 24- to 48-hr duration with slow rewarming in human brain injury showed indications of neurologic improvement and a low incidence of hypothermia-related complications. Based upon the strengths of both laboratory and clinical data, a multicenter (nine centers), randomized, prospective trial testing moderate systemic hypothermia in patients with severe brain injury has been organized. This trial, funded by National Institutes of Health, National Institute of Neurological Disorders and Stroke, began on October 20, 1994. Five hundred patients are to be treated in an intent-to-treat protocol using standard management at normothermia versus standard management at hypothermia. The trial is designed to detect an absolute shift of 12% in the percentage of patients achieving satisfactory outcome (good recovery/moderate disability) at a power of 85% at 6 months postinjury. The efficacy of hyperbaric oxygen administered every 8 hrs for 1-hr duration for a 2-wk period has also been tested in patients after severe brain injury. While the mortality rate was reduced in the treated group, the percentage of favorable outcomes was unchanged. Further studies are in progress. PMID- 7496758 TI - Airway management and mechanical ventilation. AB - The care of the acutely head-injured patient involves rapid evaluation and correction of hypoxia with appropriate airway management and treatment of associated problems. The primary focus is correction of underlying pathophysiology and prevention of secondary brain injury. Anesthetics and muscle relaxants are used to control intracranial dynamics and maintain systemic and cerebral perfusion. Airway management requires prompt intervention and definitive control, while protecting the cervical spine. Preparation for the possibility of failed intubation is important. PMID- 7496759 TI - Fluid management in patients with traumatic brain injury. AB - Movement of water between the brain and the intravascular space is dependent on osmotic gradients, which may be established by the acute administration of either hyper- or hypo-osmolar solutions. Mannitol, a hypertonic crystalloid solution, is commonly used to decrease brain water content and reduce intracranial pressure (ICP). Hypertonic saline solutions also decrease brain water and ICP while temporarily increasing systolic blood pressure and cardiac output. Hypo-osmolar solutions, such as 5% dextrose in water, reduce serum sodium and increase brain water and ICP. Colloid solutions exert little influence on either variable. Fluid restriction minimally affects cerebral edema and, if pursued to excess, may result in episodes of hypotension, which may increase ICP and are associated with worse neurologic outcome. Although there is no single best fluid for patients with traumatic brain injury, isotonic crystalloids are widely used and can be justified on a scientific basis. PMID- 7496760 TI - Hemodynamic management of brain-injured patients. AB - Head-injured patients require maintenance of systemic hemodynamics as well as attention to cerebral hemodynamics. Most head-injured patients have increased metabolic oxygen consumption, mild hypertension, and increased cardiac indices. Assessment of regional perfusion, difficult in many patients, includes monitoring of urinary output. In head-injured patients, especially those with multiple injuries, the two most important goals are preservation of cerebral perfusion pressure (mean arterial pressure minus intracranial pressure) and maintenance of systemic oxygen availability (cardiac index times arterial oxygen content). PMID- 7496761 TI - Nutrition in the head-injured patient. AB - Neurologic injury initiates a cascade of local and systemic metabolic responses. Patients become hypermetabolic, hypercatabolic, and hyperglycemic and develop decreased immune competency and altered gastrointestinal function. Provision of an adequate supply of nutrients is associated with improved outcome. Because of the high incidence of gastroparesis, enteral nutritional support of patients with acute head injury has traditionally been difficult and led to frequent use of parenteral nutrition. New methods of early enteral feeding allow a majority of these patients to receive adequate nutrition via the enteral route. Evidence indicates that early small bowel feeding of patients with acute head injury results in a decreased incidence of infections and shorter ICU stay. The roles of specific nutrients in modulating injury responses are currently being evaluated. The optimal nutritional support for improving neurologic recovery following head injury remains to be established. PMID- 7496762 TI - Causes, prevention, and treatment of post-traumatic epilepsy. AB - Post-traumatic seizures often occur after severe head injury. Acutely, these seizures complicate management of the head-injured patient by increasing intracranial pressure and causing postictal decreases in level of consciousness. In the long term, epilepsy can have a negative effect on the patient's functioning and integration into society. The more severe the head injury, the more likely that post-traumatic seizures will occur. The risk of late seizures exceeds 30% for patients with penetrating head injury, intracerebral hematoma, subdural hematoma, depressed skull fracture, or seizure within the first week after injury. Late post-traumatic seizures are treated the same as any epileptic seizures of the same type. Phenytoin and carbamazepine are effective in preventing seizures in the first week after head injury, but are not effective in preventing late seizures. Both additional antiepileptic drugs and neuroprotective agents that may lessen the damage that leads to seizures are being investigated to determine if they are effective in preventing the occurrence of post-traumatic epilepsy. PMID- 7496763 TI - Anesthesia and head trauma. AB - Anesthetic care for patients with traumatic brain injury involves an integration of cerebral resuscitation, resuscitation of other vital organs, the provision of "anesthesia," and the prevention of harmful physiologic responses to surgery. Adverse responses to surgery such as hypertension, tachycardia, coughing, and straining can increase intracranial pressure (ICP). Airway manipulations can aggravate spinal cord injury as well as increase ICP. Anesthetic agents can exacerbate hemodynamic instability, increase cerebral blood volume and ICP, and produce respiratory depression. Cerebral resuscitation during surgery resembles that in the preoperative and postoperative periods. ICP measurement and jugular venous saturation monitoring may help define end-points for cerebral resuscitation. Various anesthetic techniques and agents have distinct advantages and disadvantages. The choice of agents and techniques is determined by the nature and severity of the patient's injuries and by pre-existing medical problems. The investigation of drugs that might protect against cerebral ischemia has included anesthetic agents but none appear to be uniformly effective. Intraoperative hypothermia is also being investigated as a cerebral protectant. PMID- 7496765 TI - Pediatric issues in head trauma. AB - Injuries are the leading cause of death in children, and brain injury is the most common cause of pediatric traumatic death. Numerous mechanisms of injury are unique to the child, including walker injuries, skateboard injuries, falls, and child abuse, as well as those injuries common to all age groups. Motor vehicle accidents account for the largest number of severe injuries in children. The cerebrovascular response of the child is different from the adult's in severe head injury. The patient's resting cerebral blood flow is higher. There seems to be a higher incidence of cerebral swelling, although not as frequent as previously thought. Special attention should focus on the small systemic blood volume, temperature instability, and difficulty in intubating small children. Intracranial pressure (ICP) monitoring should be performed in patients with severe head injuries. The occurrence of increased ICP in severe head injury may be as high as 60% during the patient's course. Child abuse, as well as the "shaken baby" syndrome, is a particular problem that should be recognized and appropriately handled. The outcome should be carefully assessed in the young patient. While the injury may be the cause of a patient's abnormal development, other factors should also be considered. PMID- 7496764 TI - Medical complications in severe head injury. AB - Virtually any medical complication can occur during the post-traumatic course of an individual patient with a severe head injury (SHI). Although recovery of an individual patient can be adversely influenced by any of these medical complications, only a few selected complications have been proven to be independent predictors of an unfavorable outcome. The most frequent medical complications in SHI patients include disturbances of serum electrolytes (60%) and blood coagulation (18%), pneumonia (40%), and septicemia (10%). Hypotension is also common, occurring both prehospital and inhospital at a frequency of 29% and 22%, respectively. These factors are statistically independent predictors of poor outcome in SHI patients. It has been suggested that elimination of these frequent complications might result in a 7.5% reduction of unfavorable outcome in this patient group. PMID- 7496766 TI - Neurotrauma and trauma systems. AB - Optimal trauma care, including that for head and spinal cord injury, requires system organization and adoption throughout the United States and the world. Neurosurgeons play an essential role in system design and development in addition to treating neurotrauma patients. Areas of neurosurgical involvement include defining prehospital triage and treatment guidelines, emergency department evaluation and therapy, operative management, and active involvement in the critical care and acute hospital settings. Collaboration among all members of the trauma team is essential to ensure the best possible outcome for patients with traumatic injuries. PMID- 7496767 TI - Neuroimaging in head injury. AB - This article reviews the neuroradiologic manifestations of acute nonpenetrating head injury. The pathophysiology of the various types of neurotrauma is correlated with the imaging features. Computed tomography (CT) remains the initial imaging modality of choice, allowing rapid diagnosis of the majority of injuries. Magnetic resonance imaging is useful in the subacute and chronic stages (particularly in a patient whose initial CT scan is unremarkable) to assess brainstem and white matter shearing injuries. PMID- 7496768 TI - Pharmacologic therapy for traumatic brain injury: experimental approaches. AB - An important feature of traumatic brain injury is that much of the ultimate damage appears to occur in a delayed or secondary fashion. Although the exact timing of these secondary sequelae has yet to be elucidated, recent experimental evidence suggests that an extended window of opportunity exists during which various forms of therapy appear to be efficacious. Moreover, new therapies have been developed which can be targeted at distinct pathophysiologic aspects of brain trauma. This article summarizes recent efforts to define secondary mechanisms of brain trauma and review the development of therapeutic strategies for reversing these deleterious events. PMID- 7496770 TI - Medical management of severe head injury: present and future. AB - The comparative efficacy of various treatment algorithms in improving outcome from severe head injury (SHI) has never been tested in a prospective, randomized, controlled trial. Indeed, there are few hard data on the influence on outcome of most of the individual treatment modalities used alone. The medical management algorithm presented here is an exercise in evaluating the strength of what studies do exist and attempting to balance the relative risk/benefit ratios of the various treatment modalities. This algorithm, based on the information contained in this issue of New Horizons, divides the patient's course into two segments based on the insertion of an intracranial pressure (ICP) monitor. Before the establishment of ICP monitoring, based on the devastating effects of secondary insults on the injured brain, the main emphasis should be on full resuscitation of the patient. Any "prophylactic" treatment of the intracranial injury that has the potential of interfering with full resuscitation (e.g., mannitol) or inducing secondary ischemic insults (e.g., hyperventilation) should be reserved for the specific instance of evidence of herniation or neurologic deterioration; if such deterioration should occur, however, it should be promptly treated. Following computed tomography imaging and any necessary surgical procedures, and ICP monitor should be inserted and treatment directed specifically toward controlling ICP and maintaining a cerebral perfusion pressure > or = 70 mm Hg. An algorithm for treating intracranial hypertension is presented, based on the successive application of effective agents with increasing attendant risks. Outside of the burgeoning pharmacologic approaches to the injured brain, the future of the management of SHI involves: a) subjecting the various protocols and treatment modalities presently in use to prospective, randomized, controlled trials in order to formally establish their utility; b) developing organized, regionalized trauma care systems which facilitate the universal delivery of the level of care necessary to effectively apply today's head injury management protocols; and c) furthering our development of targeted therapy in treating SHI. Targeted therapy involves recognizing and understanding the various pathophysiologic processes that occur in the injured brain over the acute course of treatment and the responses of these processes to various treatment modalities. Such processes include vasogenic and cytotoxic edema, increased cerebral blood volume, altered cerebrovascular autoregulation, vasospasm, etc.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 7496769 TI - Pharmacologic therapy: promising clinical investigations. AB - The identification of ischemia as a central mechanism of cell injury within the nervous system has resulted in a commonality of interests of neurosurgeons, neurologists and basic scientists interested in head injury, subarachnoid hemorrhage and stroke. This, in turn, has led to a certain synergy between the development of new agents and the availability of interested and committed investigators so that these agents can be tested under appropriate clinical conditions. In head injury, major sources of focus have been the membrane damage resulting from the free-radical cascade and the disruption in cellular ionic homeostasis resulting from the excitotoxic effects of pathologic release of amino acid neurotransmitters. At present, the final analyses of phase III trials of the antagonism of the initiation and propagation of the free-radical cascade by tirilazad and polyethylene glycol-bound superoxide dismutase are nearing completion. Data on the efficacy of these agents in improving outcome from moderate and severe head injury should be available within the next 6 months. In addition, worldwide trials of glutamate antagonists are presently being initiated in severe head injury, with results anticipated in late 1997 or early 1998. In subarachnoid hemorrhage, calcium has been hypothesized to play a role in mediating vasospasm-induced ischemia as well as in promoting intracellular damage. Trials of calcium-channel blockade have demonstrated a reduction in mortality, but no improvement in the quality of survival over other methods of managing these patients.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7496771 TI - Detection of the Machado-Joseph disease/spinocerebellar ataxia three trinucleotide repeat expansion in families with autosomal dominant motor disorders, including the Drew family of Walworth. AB - Affected members of 63 families with a variety of autosomal dominant late onset cerebellar ataxias (ADCA), and 29 patients with similar phenotypes but no affected relatives, were investigated for the trinucleotide (CAG) repeat expansion described in Japanese families with Machado-Joseph disease (MJD). This disorder had previously been shown to map to the region of chromosome 14 which also contains a locus causing ADCA in French families, spinocerebellar ataxia 3 (SCA3). The MJD/SCA3 mutation was identified in nine families with ADCA type I, and a further family in which affected members had parkinsonism, peripheral neuropathy, dystonia, and spasticity, but little evidence of cerebellar disease. Only one of the 10 families was British (the Drew family of Walworth); the others originated from India, Jamaica, Ghana, Brazil and France. There was no single clinical feature which distinguished patients with the MJD/SCA3 mutation from those with the CAG expansion on chromosome 6 (SCA1) or ADCA type I families with no known mutation. The CAG repeat length ranged from 13-41 copies on normal chromosomes and 62-80 copies on affected chromosomes. There was a significant inverse correlation between age of onset of symptoms and repeat length, but no significant effect of parental sex on repeat length or age of onset in offspring. DNA analysis for the MJD/SCA3 mutation is useful for diagnosis in patients with familial ataxic or extrapyramidal syndromes, and will aid genetic counselling in these disorders. PMID- 7496772 TI - Progressive myoclonic ataxia associated with coeliac disease. The myoclonus is of cortical origin, but the pathology is in the cerebellum. AB - We report four patients with a progressive myoclonic ataxic syndrome and associated coeliac disease. The onset of the neurological syndrome followed the gastrointestinal and other manifestations of coeliac disease while on a gluten free diet, in the absence of overt features of malabsorption or nutritional deficiency. The condition progressed despite strict adherence to diet. The neurological syndrome was dominated by action and stimulus sensitive myoclonus of cortical origin with mild ataxia and infrequent seizures. Plasmapharesis and immunosuppressive treatment were tried in two patients but were not beneficial. Post-mortem examination of the brain in one case showed selective symmetrical atrophy of the cerebellar hemispheres with Purkinje cell loss and Bergmann astrocytosis, and with preservation of the cerebral hemispheres and brainstem. Coeliac disease should be considered in the differential diagnosis of all patients presenting with a progressive myoclonic ataxic syndrome. PMID- 7496773 TI - Familial progressive supranuclear palsy. Description of a pedigree and review of the literature. AB - We describe a family with autosomal dominant progressive supranuclear palsy (PSP) involving five generations which was confirmed in one patient. The proband presented with progressive slowness at age 53 years, followed by ocular palsy, loss of balance, axial dystonia, dysphagia and dysarthria, and died at age 59 years. Neuropathological examination revealed moderate numbers of neurofibrillary tangles without prominent senile plaques in the cortex, and neuronal loss, gliosis and moderate to severe accumulation of tangles in the basal ganglia and brainstem. Other affected relatives, including the proband's sister, father, paternal uncle, and other members of earlier generations presented with non characteristic akinetic syndromes, which progressed towards more typical PSP only after several years of disease. A review of the literature revealed six other families with neurodegenerative disorders associated with pathological findings compatible with PSP in at least one member. The clinical symptoms varied greatly between individuals in these families. The pattern of inheritance seems compatible with autosomal dominant transmission, although other patterns of transmission could not be excluded. We conclude that there is an autosomal dominant form of PSP and that the number of hereditary cases may be greater than previously thought. The rarity of familial cases of PSP could be attributed to diagnostic problems, including lack of recognition of atypical cases and death of the gene carriers before the age of appearance of the clinical symptoms. Large families with hereditary PSP could provide an adequate point of departure for investigation of the gene defect responsible for this disease. PMID- 7496774 TI - Motion and shape perception in cerebral akinetopsia. AB - Motion cues serve many purposes in primate vision. Consequently, akinetopsia, a defect of movement perception due to cerebral lesions, would be expected to comprise a range of motion-related defects. To address this issue we explored further the perceptual profiles in akinetopsic subject L.M. who has motion perception deficits due to extensive bilateral lesions of the dorsolateral visual association cortex that spare primary visual cortex, area V1. We used several different experimental procedures. Using random-dot motion stimuli, we showed that L.M. can still perceive global coherent motion and discriminate motion direction, yet these abilities fail even at moderate levels of background noise. L.M. also viewed a two-frame apparent (phi) motion stimulus known as a Ternus display. Her performance on this test suggests defective 'long-range' apparent motion mechanisms. These results are clearly in agreement with previous reports. Additional experiments showed that L.M. can still perceive 2-D shape and 3-D structure-from-motion (SFM). However, like motion direction discrimination, these abilities broke down at moderate levels of moving and stationary noise. Surprisingly, L.M. also had trouble perceiving 2-D shapes defined by non-motion signals including 'on' and 'off' transients, dynamic and static binocular disparity, and static texture cues. Our findings highlight the role of the visual association cortex in extracting salient information from noise. PMID- 7496775 TI - Double dissociation of memory capacities after bilateral occipital-lobe or medial temporal-lobe lesions. AB - Memory for recently encountered information can be reflected in conscious recall and recognition of that material, or in facilitated reprocessing of that material, an effect known as repetition priming. Repetition priming may be perceptual (form-based) or conceptual (meaning-based). A patient with bilateral occipital-lobe lesions (L.H.) and a patient with bilateral medial-temporal lobe lesions (H.M.) showed a double dissociation between visuoperceptual priming (impaired in L.H. and intact in H.M.) and visual recognition memory (intact in L.H. and impaired in H.M.). L.H. showed intact conceptual priming for visually presented words; his pattern of impaired visuoperceptual priming and intact conceptual priming is the reverse dissociation to that observed in prior studies of patients with Alzheimer's disease, in whom occipital cortices are relatively spared. These double dissociations suggest that a memory system localized to the occipital lobe mediates visuoperceptual priming effects, and that this system is independent of neural circuits mediating conceptual priming effects, and independent of the limbic-diencephalic system supporting conscious recognition of recently encountered information. PMID- 7496776 TI - Weight judgment. The discrimination capacity of a deafferented subject. AB - A weight discrimination study was undertaken to test (i) the capacity of controls and a deafferented subject (deprived of large sensory myelinated fibres from nose down), to discriminate weights with and without vision; (ii) the capacities of observers to discriminate weights while watching the deafferented and control subjects' lifting movements; (iii) the contribution of supplementary sources of sensory information (e.g. vestibular afferents) to the deafferented subject's discrimination capacity. With vision, G.L.'s liminal discrimination of weights was similar to that of the controls. In contrast, precluding vision impaired massively, but not completely, G.L.'s discrimination capacity, so emphasizing the importance of visual kinaesthetic cues in G.L. and incidently the importance of large myelinated sensory function in weight discrimination in controls. Kinematics recordings of G.L.'s lifting movements with vision revealed a significant correlation between weight and peak velocity of the lifting movement. This reflects a specific strategy used by G.L. to generate movements, allowing her to judge the weight of a lifted object visually. Peak velocity rather than amplitude of movement appears to be the main cue for G.L. since there was a lack of correlation between amplitude and weight lifted. For controls, none of the correlations (weight versus amplitude or weight versus velocity) was significant, whether vision was available or not. When watching G.L.'s lifting performance, external observers were able to use similar cues to establish their judgments, but they were far less accurate in doing so when watching control subjects. This suggests that controls were using a strategy different from G.L.'s.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7496777 TI - The perception of head and neck angular displacement in normal and labyrinthine defective subjects. A quantitative study using a 'remembered saccade' technique. AB - The detection of horizontal angular displacement of the head or trunk was assessed in 13 normal subjects and three labyrinthine defective patients. Discrete rotational displacements to the whole body (vestibular stimulus, VS), the trunk whilst keeping the head earth-stationary (cervical stimulus, CS) or the head upon the stationary trunk (combined VS+CS in passive and active mode) were delivered. Amplitudes ranged from 10 degrees to 40 degrees with peak velocities from 5 degrees to 40 degrees/s. Response was quantified as saccadic gain (final position of the eye/amplitude of the rotational stimulus) in a 'remembered saccade' task. it was found that normal subjects show different accuracy according to the sensory channel investigated. If stimulus amplitude was limited (20-30 degrees), gain dropped as rotational velocity decreased during VS but not in conditions including cervical stimulation (CS or VS+CS). Responses combining cervical and vestibular stimuli (VS+CS) were closer to unity gain and showed less scatter than isolated VS and CS. In the labyrinthine-defective patients, VS yielded no structured response but all responses involving cervical stimulation were identical to those of normal subjects when analysed both as a function of stimulus amplitude and velocity. Neither in normal nor in labyrinthine-defective subjects were there significant differences in the ability to detect head angular displacement between passive or active head turns (passive or active VS+CS). The following conclusions can be made. (i) 'Remembered saccade' techniques can be used to investigate cervico-vestibular perception. (ii) The 'high pass' characteristic of the response during VS agrees with the dynamic sensitivity of the vestibular nerve signals. Cervical experiments confirmed that neck responses are position-driven (i.e. 'tonic'). (iii) The detection of head turns is only slightly more accurate when vestibular and cervical signals are combined, but the main input controlling this perception is provided by neck afferents. No specific function for 'efference copy' was apparent in these experiments. (iv) There is no significant change for detecting head turns (on the trunk) in labyrinthine defective patients. (v) The lack of detectable changes in cervical tasks in labyrinthine-defective patients does not support the view that the potentiation of the cervico-ocular reflex (COR) observed in these patients is due to enhanced sensitivity of the neck afferent system. PMID- 7496778 TI - Cortical control of vestibular-guided saccades in man. AB - Memory-guided saccades, made to a remembered location to which gaze was directed before a passive body rotation (i.e. with a vestibular input), were electro oculographically recorded in 24 patients with various cortical lesions and in 18 control subjects. Anticipation and latency, direction errors and accuracy of the first saccade, stability of eye position in darkness and final eye position were quantified. Patients were divided into small groups, each with lesions affecting one of the following cortical areas: left or right frontal eye field (FEF), left or right prefrontal cortex (area 46 of Brodmann) (PFC), left supplementary eye field (SEF), left or right posterior parietal cortex (PPC) and right parieto temporal cortex (PTC). There were some abnormalities in the results of the right FEF group, concerning anticipation, direction errors and latency of the first saccade, but no abnormality in the accuracy of the first saccade or of the final eye position. Results in the left FEF group were normal. Accuracy of the first saccade was impaired in the SEF group, bilaterally. Final eye position was also inaccurate in the SEF group. In both PFC groups, significant and, in general, bilateral abnormalities existed for all tested parameters. Accuracy of the first saccade was impaired in the PTC group, leftwards. In contrast, the results in both PPC groups were not significantly different from those of control subjects. Our results suggest that (i) the PFC is involved in the memorization of saccade goals probably encoded in spatiotopic coordinates; (ii) the SEF, but not the FEF, is involved in the control of accuracy of these vestibular-derived goal-directed saccades; (iii) the PTC (i.e. the vestibular cortex), but not the PPC, is involved in the control of such saccades. Therefore, a cortical network different from that involved in the control of memory-guided saccades made to visual targets, with only the PFC in common, could control vestibular-derived goal directed saccades. PMID- 7496779 TI - Transcranial magnetic stimulation over the posterior cerebellum during visually guided saccades in man. AB - Recent intensive neurophysiological experiments in the monkey have demonstrated the function of the cerebellum in the control of saccadic eye movements. Microstimulation studies on monkeys have determined that the cerebellar cortex, which is specifically involved in the control of saccades, is located in vermal lobules VIc and VII. The Purkinje-cell axons arising from this vermal area terminate almost exclusively in an ellipsoidal region which protrudes dorsocaudally from the fastigial nucleus. Saccade-related cells in the fastigial nucleus are located exclusively in the ellipsoidal region. Microstimulation of the vermis modulated the activity of saccade-related cells in the fastigial nucleus, and then produced dysmetric saccades. In this study, we investigated effects of cerebellar stimulation on saccade metrics in man using a transcranial magnetic stimulation (TMS) device. Focal TMS was applied over the posterior cerebellum at an area approximately 7 mm lateral and caudal to the inion during horizontal visually guided saccades in six normal subjects. The TMS device was triggered after the onset of saccades with a latency of 0, 20, 40 or 60 ms. We investigated the effect of TMS on the amplitude and velocity of saccadic eye movements. For visually guided saccades directed contralateral to the stimulation side, TMS of the posterior cerebellum with the latency of 0 ms produced hypometric saccades followed by corrective saccades. Transcranial magnetic stimulation with latencies of 20, 40 and 60 ms had no effect on saccade metrics. On the other hand, for ipsilateral saccades, TMS with latencies of 0, 20 and 40 ms produced hypermetric saccades followed by postsaccadic drift.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7496780 TI - Non-decussating retinal-fugal fibre syndrome. An inborn achiasmatic malformation associated with visuotopic misrouting, visual evoked potential ipsilateral asymmetry and nystagmus. AB - We report a newly identified syndrome in which nasal retinal fibres fail to decussate due to the inborn absence of an optic chiasm. Visual evoked potential (VEP) assessment and neuro-opththalmic evaluation in two unrelated, non-albino children revealed the unusual visual pathway anomaly in the form of misrouted retinal-fugal projections. Monocular VEP responses across the occiput, regardless of stimulus mode (full- or partial-field pattern onset, pattern reversal, luminance flash or high temporal frequency luminance flicker) showed unequivocal evidence of pathological VEP ipsilateral asymmetry. Marked attenuation of primary visual evoked responses from the occiput contralateral to the eye of stimulation, indicative of aberrant contralateral retinal-fugal projections, was confirmed by MRI which depicted the remarkable achiasmatic condition. MRIs and neurological evaluation also confirmed the absence of accompanying congenital or acquired brain malformations or anomalies. Ophthalmic evaluation revealed that both achiasmatic children had reduced distance acuity for age, alternating esotropia, torticollis, head tremor and ocular motor instability; visual fields were normal. Eye movements were also monitored and indicated congenital nystagmus waveforms in the horizontal plane; see-saw nystagmus was observed in the the horizontal plane; see-saw nystagmus was observed in the vertical and torsional planes. The age range of the two children during evaluation and follow-up, over a 6-year period, was about 4-15 years. Comparisons of VEP responses from age-matched normal, albino and idiopathic congenital nystagmus controls, recorded under the same VEP test conditions, were also performed. In contrast to the achiasmatic ipsilateral inter-ocular asymmetry, the albinos showed the expected monocular VEP topography pattern of contralateral asymmetry. Also as expected, VEP profiles from the normal controls and those with congenital nystagmus, evinced no aberrant asymmetry patterns. In general, the results indicate that the VEP misrouting protocol is indispensable for the non-invasive electrophysiological detection and differential diagnosis of optic pathway mutations and may well identify individuals with purported idiopathic congenital nystagmus or albinism that are, in fact, achiasmatic. PMID- 7496781 TI - Activity-dependent changes in impulse conduction in normal human cutaneous axons. AB - The present study was undertaken to determine if the axonal hyperpolarization produced by a brief train of impulses would impair neural transmission in cutaneous afferents of normal human subjects (n = 25). To assess changes in axonal excitability, a submaximal test stimulus was conditioned by a train of 10 supramaximal stimuli at 200 Hz. This produced a depression in excitability lasting up to 100 ms, demonstrable at nodes of Ranvier remote from the site of stimulus application, and probably due to activation of a slow K+ conductance. The effects of this change in excitability on neural transmission were assessed using a supramaximal test pulse. This revealed small but significant activity dependent decreases in amplitude at conditioning-test intervals up to 20 ms and increases in latency at intervals up to 70 ms. Both the amplitude decrease and the latency increase were greater the longer the conduction distance. The reduction in amplitude of the compound sensory potential could be explained by temporal dispersion due to the increase in latency. It is concluded that, at the nodes of normal cutaneous afferents, the safety margin for impulse generation is sufficiently high that the activity-dependent hyperpolarization does not produce conduction block. It is likely that the previously described reductions in the amplitude of the compound sensory action potential in response to brief trains of stimuli were due to dispersion of the volley, not conduction failure, and that conduction failure does not occur in normal cutaneous axons solely by activation of slow K+ conductances.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7496782 TI - A mechanism for ectopic firing in central demyelinated axons. AB - Intracellular recordings of ectopic activity were made from demyelinated axons in rat dorsal columns using in vitro techniques. On-going bursts of discharges were observed in some axons, and these were sometimes superimposed upon slow depolarizing potentials. These intracellularly recorded, depolarizing potentials were strongly reminiscent of the slow negative potentials previously recorded extracellularly from the periaxonal region of normal myelinated fibres following potassium loading of this space. Also, in previously silent demyelinated axons, ectopic activity could sometimes be induced by brief periods of high frequency stimulation applied remotely from the lesion. The generation of the ectopic activity can be explained in terms of an artificially increased periaxonal concentration of potassium which may lead to the opening of internodal potassium channels and an inward potassium current. We suggest that a raised periaxonal potassium concentration may occur within compartments in demyelinating lesions, and that it can result in the generation of ectopic impulses. PMID- 7496784 TI - AIDS-associated vacuolar myelopathy. A morphometric study. AB - The post-mortem pathology in 20 spinal cords of human immunodeficiency virus (HIV) infected patients with vacuolar myelopathy was quantified by evaluating (i) the intensity of myelin change, vacuolation and macrophage density; and (ii) the areas of white matter covered by each feature. Severity scores were constructed for (i) the anterior, lateral, and posterior white matter columns; (ii) each level of spinal cord; and (iii) the whole spinal cord [Cord Total and Cord Average Severity Scores (CTSS, CASS)]. Astroglial activation was scored separately. In 14 cords with mild-moderate vacuolar myelopathy (CASS = 23-259), macrophages were the most prominent pathological feature, and level severity scores were higher at mid-thoracic than cervical levels (P = 0.009). In six cords with severe vacuolar myelopathy (CASS = 396-614), vacuolation, demyelination and macrophages were equally evident and thoracic and cervical level severity scores were similar. The most severe lesions showed evidence of clearing of macrophages from the collapsed centres. A clinical lower limb score correlated with the anterior (P = 0.03) and lateral (P = 0.04) column total scores and with the CTSS (P = 0.04) in the nine patients who had had both myelopathy related disability and all cord levels available. There was no significant longitudinal gradient in score severity in the posterior, lateral or anterior columns and no evidence of a dying-back phenomenon. There was no evidence of Wallerian degeneration occurring as a primary process. Astroglial activation did not correlate with the severity or duration of the vacuolar myelopathy. Detection of HIV p24 antigen in the spinal cord related to the local presence of multinucleated giant cells and to antigen expression in the brain but not with the severity of vacuolar myelopathy. The pathology in vacuolar myelopathy appeared to start in the mid-low thoracic cord, with increasing rostral involvement as the disease became more severe. The relative prominence of macrophages in mild-moderate lesions suggests they may be involved early in the pathogenesis of vacuolar myelopathy. PMID- 7496783 TI - Non-Hodgkin malignant lymphomas and peripheral neuropathies--13 cases. AB - Non-Hodgkin's malignant lymphomas (NHML) are malignant lymphoid proliferations which may be of B or T cell type. Thirteen observations of an association between peripheral neuropathy and B type NHML are reported. None of the cases had evidence of meningeal propagation or neurotoxicity from chemotherapy. The NHML were classified according to the Working Formulation and Kiel classifications. The various mechanisms of peripheral neuropathy in these cases were split into four broad groups. Group I consisted of four cases in which the peripheral nerve lesions were directly linked to a propagation of malignant cells into the peripheral nervous system; this was revealed by autopsy and/or nerve biopsy. Malignant B cell proliferation was demonstrated in three out of four of these cases by immunolabelling of the infiltrates. Group II included three patients whose serum contained a monoclonal immunoglobulin (IgM) with antimyelin activity, and two who had pathological IgM deposits in endoneurial connective tissue. Group III comprised two cases. The immune dysfunction of the NHML was responsible for a Guillain-Barre syndrome in one, and for a chronic inflammatory demyelinating polyneuropathy in the other. Group IV included two patients in whom the mechanism of the peripheral neuropathy, although almost certainly directly related to the NHML, could not be determined beyond doubt. The peripheral neuropathy might have been a result of a paraneoplasic process or, possibly, an undetected lymphomatous invasion of nervous tissue. All these cases of clinically diverse peripheral neuropathy, which either occurred before the discovery of the haemopathy or arose as complications of it, are discussed along with similar observations reported in the literature. Immunolabelling of lymphomatous proliferations and nerves is now of considerable value for classifying and indicating the exact aetiology of the peripheral neuropathy. It can also detect pathogenic consequences of any associated monoclonal dysglobulinemia. In any event, a direct link between the peripheral neuropathy and NHML represents an indication for intensification of specific chemotherapy, which in some of our patients led to significant regression of the peripheral neuropathy. Nonetheless, in some cases, the link between peripheral neuropathy and NHML could not be established with certainty. Long-term follow-up is essential in such cases. The present results show the importance of a case by case study of patients with NHML and peripheral neuropathy. PMID- 7496785 TI - Non-neural-specific T lymphocytes can orchestrate inflammatory peripheral neuropathy. AB - Neural-specific T lymphocytes are held to play a pathological role in inflammatory peripheral nerve disorders such as the Guillain-Barre syndrome (GBS) and chronic inflammatory demyelinating polyneuropathy (CIDP). Here, non-neural specific T-cell-mediated inflammation was studied in peripheral nerves in Lewis rats by systemic transfer of ovalbumin-specific activated T cells followed by intraneural injection of ovalbumin. Rapid endoneurial perivenular infiltration of alpha beta T cells and ED1+ macrophages occurred with ovalbumin injection following transfer of 2 x 10(6) T cells. This cellular infiltration and accumulation produced marked increases in blood-nerve barrier (BNB) permeability. In contrast, control casein injections produced neither significant cell accumulation nor BNB permeability changes. Transfer of a higher number of T cells (5 x 10(6)) induced severe Wallerian degeneration and nerve conduction failure in ovalbumin injected nerves. Fewer T cells (5 x 10(5)) induced conduction block and mild demyelination which were markedly augmented by systemic cotransfer of anti myelin immunoglobulin. This study demonstrates that activated T cells of non neural specificity can accumulate in peripheral nerve, produce dramatic changes in BNB permeability and with intravenous anti-myelin antibody orchestrate primary demyelination or axonal degeneration in a dose-dependent fashion. PMID- 7496787 TI - Tumoural parietal lobe epilepsy. Clinical manifestations and outcome in 34 patients treated between 1934 and 1988. AB - Between 1934 and 1988, 34 patients with tumoural parietal lobe epilepsy were treated surgically at the Montreal Neurological Institute (MNI). Fifteen had right-sided and 16 left-sided resections. The remaining three patients had biopsies only. Follow-up ranging from 1 to 40 years (mean 12.3 years) was available for 28 patients. Seventy-five percent (21 out of 28) became seizure free or had rare seizures. Permanent post-operative sensory deficits were described in 12% of patients. An aura was described by 79%. 62% had somatosensory symptoms, contralateral to the epileptogenic region in all but one. Visual illusions (12%), aphasia (9%) and disturbances of body image (6%), were much less common at the beginning of the attacks. Intra-operative cortical stimulation reproduced the habitual aurae in 10 out of 25 (40%) of the patients. The clinical manifestations suggested different spread patterns: 21% had tonic posturing of the extremities, 82% focal clonic activity, 15% head deviation, 9% automatisms and 6% difficulty speaking. Eleven (32%) had Todd's paralysis and 18% postictal dysphasia. Almost half the patients had impaired two-point discrimination in contralateral fingers; two of these also had impaired stereognosis, but only one had astereognosis without coexisting primary cortical sensory deficit. Review of this, now historical, series shows that parietal lobe tumours can be resected with good control of previously intractable seizures. Such an approach is preferable to postponing resection until the lesion is shown to increase in volume. PMID- 7496786 TI - Periventricular and subcortical nodular heterotopia. A study of 33 patients. AB - Grey matter heterotopias, demonstrated by MRI, may present with a broad spectrum of clinical severity. We have studied 33 patients with periventricular nodular heterotopias (PNH); 19 (58%) had unilateral and 14 (42%) bilateral lesions. Thirteen of the 19 patients (68%) with unilateral subependymal nodules of grey matter had, in addition, unilateral focal subcortical heterotopias (SNH), comprising 39% of the entire group. Most had normal intellectual and motor function but some presented with mild mental retardation and neurological deficits. Recurrent seizures were described in 82%, mainly partial attacks with temporo-parieto-occipital auras. Nodular heterotopias led to unilateral or bilateral independent temporal epileptic discharges in 47% of epileptic patients with PNH alone and in 61% of those who had SNH in addition. Extratemporal or multilobar, unilateral or bilateral interictal spiking was present in 10 other patients (36%). Two first degree relatives of patients with seizures were affected but had no seizures, three were investigated for other apparently unrelated neurological symptoms: memory impairment, vertigo or transient ischaemic attacks in one person each. Contiguous ovoid nodules of grey matter, symmetrically lining both lateral ventricles, were described in nine patients. Seven of them were female, including four with familial incidence of PNH. Such lesions may explain the familial occurrence of epilepsy in some families. Seven patients underwent anterior temporal resection: two patients with unilateral subependymal and focal subcortical heterotopias were seizure free or significantly improved. Four patients, three with PNH alone and one with additional subcortical nodules, did not improve significantly after surgery. The remaining patient was followed for less than 6 months. PMID- 7496788 TI - The Simon effect and attention deficits in Gilles de la Tourette's syndrome and Huntington's disease. AB - Tourette's syndrome and Huntington's disease have long been clinically associated with attentional deficits. In this study, we aimed to determine the nature and quantify the extent of such deficits. A technique was devised to ascertain the efficiency with which Tourette's syndrome and Huntington's disease patients could shift and direct attention away from naturally expected stimulus-response (S-R) linkages. This was done by varying the relationships formed between stimulus and response location. Attentional efficiency was indicated by relative speed of responding to relevant (congruent) and irrelevant (incongruent) stimuli, in a paradigm developed from the Simon effect. There were five conditions progressively increasing in complexity. The stimuli consisted of left and right pointing arrows and, in some cases, various conditionality manipulations were also employed, such that in the presence of a certain symbol (i.e. 'x') the nature of the response had to be reversed, whereas in the presence of an alternative symbol (i.e. '='), the response was compatible with the direction of the arrow. As predicted, Tourette's syndrome and Huntington's disease patients, regardless of medication or depression status and unlike controls, were particularly disadvantaged in responding to various conflicting S-R configurations. Tourette's syndrome and Huntington's disease patients may experience difficulties in making attentional shifts, or in inhibiting inappropriate responses; they may also be more susceptible (than controls) to the conflict that can arise when the spatial code formed for the stimulus is irrelevant for selecting the appropriate response. We conclude that our findings support the notion that cognitive deficits in Tourette's syndrome and Huntington's disease may stem from abnormalities of the major pathways interconnecting the basal ganglia and the frontal lobes. PMID- 7496789 TI - Transcranial Doppler and cardiovascular responses during cardiovascular autonomic tests in migraineurs during and outside attacks. AB - The transcranial Doppler (TCD) and cardiovascular responses to established tests of autonomic function were studied in 50 migraineurs (23 of whom experienced aura and 27 who did not) and 30 healthy control subjects. In addition to the measurements outside attacks, 10 migraineurs were also tested during unilateral attacks of migraine without aura. Transcranial Doppler examinations of middle cerebral artery (MCA) blood velocity showed no differences between migraineurs and healthy subjects and no difference between migraineurs experiencing an attack and outside an attack when examined in response to a head-up tilt test, a cold pressor test and a Valsalva manoeuvre. The cardiovascular reflexes in response to the Valsalva manoeuvre suggested a mild parasympathetic hypofunction in migraineurs, both those with and those without aura. On the basis of the present results and previous findings by others, we conclude that mild parasympathetic hypofunctioning with preserved sympathetic functioning is an established finding in migraine both with and without aura. PMID- 7496790 TI - Prolonged muscular flaccidity after stroke. Morphological and functional brain alterations. AB - Patients with a motor deficit due to ischaemic stroke usually develop muscular spasticity, but in some cases they may remain with a prolonged muscular flaccidity which impairs their recovery. Little is known about the causes of these two different functional outcomes. We correlated CT/MRI and 99mTc HM-PAO SPECT with clinical findings in 42 patients at a mean time interval of 3 months after stroke. The patients were divided into two cohorts with either flaccid (prolonged muscular flaccidity) or spastic (muscular spasticity) hemiparesis. Although patients with prolonged muscular flaccidity had a greater motor deficit, the mean structural volume of the ischaemic lesion was similar to that of the muscular spasticity cohort. There was a significantly higher prevalence of structural involvement of the lentiform nucleus in prolonged muscular flaccidity cases. Relative perfusion in the lentiform nucleus, thalamus and contralateral cerebellar hemisphere was significantly lower in prolonged muscular flaccidity than in muscular spasticity patients. A subgroup with only subcortical structural lesions also showed significantly lower relative perfusion in the ipsilateral frontal association areas. A primary involvement of the lentiform nucleus by the structural lesion seems to be crucial for the persistence of flaccidity after stroke. However, cerebral blood flow (CBF) changes in other structurally intact regions indicate their additional role. It is likely that both subcortical cortical loops involved in motor control, i.e. cortex-basal ganglia-thalamus cortex and cortex-pons-cerebellum-thalamus-cortex, are more widely and more severely affected in patients with prolonged muscular flaccidity. PMID- 7496791 TI - Effects of stimulus rate on regional cerebral blood flow after median nerve stimulation. AB - The primary motor cortex and supplementary motor area (SMA) are purportedly involved in the generation of the P22 and N30 components of somatosensory evoked potentials (SEPs) evoked by electrical stimulation of the median nerve at the wrist. We used regional cerebral blood flow (rCBF) measurements and PET in 10 normal subjects to study the cerebral areas activated by median nerve electrical stimulation. PET scans were performed with the subjects at rest and during stimulation of the right median nerve at frequencies of up to 20 Hz. Stimulation evoked a single focus of activation in the primary somatosensory area (SI). An increase of rCBF in this area was linearly correlated with stimulus frequencies of up to 4 Hz and then reached a plateau. The SMA was not significantly activated by stimulation at any of the frequencies tested. In contrast to the SI, the SMA showed no trend toward a correlation between the rCBF changes and the stimulus repetition rate. In order to achieve maximal resolution in the sensorimotor cortex, regions of interest were placed in individual co-registered MRI-PET images on both sides of the central sulcus. There was no significant increase of rCBF in the crown of the precentral gyrus. These results suggest that a contribution of the primary motor cortex and the SMA to the generation of the P22 and N30 components of SEPs is unlikely. Consequently, functional clinical interpretations derived from P22 or N30 abnormalities must be reconsidered. PMID- 7496792 TI - Infarcts in the anterior choroidal artery territory. PMID- 7496793 TI - Motoneurons innervating guinea pig perineal muscles are sexually dimorphic in size but not number. AB - Sexual differentiation occurs prenatally in guinea pigs but extends into the postnatal period in rats. Steroids affect the development of two motoneuron nuclei of the rat lumbar spinal cord that innervate sexually dimorphic perineal muscles. The spinal nucleus of the bulbocavernosus (SNB) innervates the bulbocavernosus (BC) and levator ani (LA) muscles while the dorsolateral nucleus (DLN) innervates the ischiocavernosus (IC). In male rats, perinatal testosterone prevents degeneration of these muscles and results in a sex difference in both motoneuron size and number in adulthood. For comparative purposes, we examined the guinea pig motoneurons innervating these muscles, as well as those innervating the retractor penis (RP) and retractor clitoris (RC), muscles that have no counterpart in rats. Injections of horseradish peroxidase localized the BC/LA and IC motoneurons of guinea pigs to discrete columns in spinal levels L6 and S1, with the BC/LA motoneurons occupying a more medial position. The RP/RC motoneurons were found in L5. Motoneuronal soma area was larger in males in all examined motor pools, as was nuclear area of BC/LA and IC motoneurons. Although raw counts suggested a sex difference in cell number in the motor columns containing BC/LA and IC motoneurons, either of two different correction procedures for split nuclei error eliminated the sex difference in cell number, emphasizing the importance of such corrections when comparing neurons of different size. PMID- 7496794 TI - Effects of different types of stress on histamine-H3 receptors in the rat cortex. AB - The role of the histamine (HA) H3 receptor (estimated by binding of [3H]N-alpha methyl HA in the brain cortex) in the response of male rats to four types of stress was evaluated: acute weak foot-shock stress (0.5 mA x 1 s x 5 times), acute intense foot-shock stress (1mA x 10 s x 5 times), acute restraint stress (60 min at 24 degrees C) and chronic variate stress. Two groups of rats received chronic treatment with a tricyclic antidepressant: amitriptyline (5 mg/kg i.p.); one group was also chronically stressed. HA-H3 receptor density rapidly decreased in acute intense foot-shock and chronic variate stress, but was unchanged in acute weak foot-shock and restraint stress. Amitripytline counteracted the chronic variate stress-induced decrease, and increased HA-H3 receptor density when chronically administered in the non-stressed control group. These results indicate that cortical HA-H3 receptor density is affected by the response to stress in function of the type, duration and intensity of the stressor. PMID- 7496795 TI - Neuropeptide Y potentiation of potassium-induced noradrenaline release in the hypothalamic paraventricular nucleus of the rat in vivo. AB - NPY is co-localised with catecholamines in the brain and periphery. Noradrenaline and NPY are present in high concentrations in the PVN of the hypothalamus, an area implicated in autonomic regulation. This microdialysis study examined whether NPY can modulate rat PVN noradrenaline release in vivo, as has been shown in vitro. Basal and K(+)-stimulated noradrenaline release was measured after i.c.v. administration of 2 nmol NPY or vehicle. No effect of NPY was observed on basal release, however a significant doubling of K(+)-induced release was observed, both 60 and 150 min following i.c.v. NPY. This raises the possibility that NPY may potentiate rather than inhibit brain noradrenaline release in vivo. PMID- 7496796 TI - Intra-VTA injections of the mu-opioid antagonist CTOP enhance locomotor activity. AB - In this paper we report on the effects of microinjections of the mu-opioid antagonist CTOP (D-Pen-Cys-Tyr-D-Trp-Orn-Thr-Pen-Thr-NH2) into the ventral tegmental area (VTA) on activity and ingestive behavior in the rat. Intra-VTA CTOP (0.015, 0.15, and 1.5 nmol per side) dose-dependently increased activity, whereas it had no effect on feeding and drinking behavior. These results are consistent with previous reports that intra-VTA injections of CTOP enhance extracellular dopamine levels in the nucleus accumbens. Furthermore, we propose a model of VTA mu-opioid mechanisms that might account for these surprising effects of intra-VTA CTOP. PMID- 7496797 TI - Do GnRH neurons express the gene for the NMDA receptor? AB - Previous studies have revealed that in several animal models, N-methyl-D,L Aspartate (NMA) stimulates LH secretion by acting at a suprapituitary site. In addition, NMDA receptor antagonists appear to block GnRH neuronal activation on the afternoon of proestrous as evidenced by the lack of c-Fos expression in the neurons and by the absence of an ovulatory LH surge. However, administration of NMA does not induce c-Fos or c-Jun expression in GnRH neurons. To better understand the effects of NMDA receptor activation on GnRH neuronal function, we examined whether GnRH neurons express the NMDA receptor in male rats, and in female rats during diestrus and proestrus, by performing double label in situ hybridization. An 35S-labeled cRNA probe for the NMDA receptor subunit (NMDAR1) was used to quantify NMDAR1 mRNA and a digoxigenin-labeled cRNA probe for GnRH was used to identify GnRH neurons. The data were quantified and expressed as grains/average cell area. In male and female rats, less than 5% of GnRH neurons expressed grain levels twice the minimum detectable level and were considered double-labeled. However, many non-GnRH neurons in the same areas as GnRH neurons expressed high levels of NMDAR1 mRNA. These results suggest that the effects of NMA on GnRH secretion are unlikely to be mediated solely by the activation of NMDA receptors on GnRH neurons. Given the widespread expression of NMDAR1 mRNA in the hypothalamus, it is possible that the stimulatory effects of NMA on GnRH neurons are indirect through activation of other neurons. PMID- 7496798 TI - Effects of naltrexone on morphine-induced tolerance and physical dependence and changes in cellular immune function in mice. AB - The effects of naltrexone on tolerance/dependence, as well as alterations in cellular immune function induced by morphine administration, were determined. Mice were rendered tolerant to and physically dependent on morphine by subcutaneous implantation of pellets containing 75 mg of morphine. Implantation of naltrexone pellets (10 mg) blocked the development of tolerance to the analgesic action of morphine, as well as the development of physical dependence. Morphine suppressed lymphoid organ weights and cellularities, and this suppression was blocked by naltrexone. B-Cell proliferation was suppressed in morphine-tolerant but not in morphine-abstinent mice, and this suppression was exacerbated by naltrexone. Morphine tolerance and abstinence were associated with suppression of IL-2 production, which was completely blocked by naltrexone. NK cell activity was not significantly affected by either morphine or naltrexone exposure. The results suggest that the effects of morphine on the immune system are at least partially mediated through opioid receptors. PMID- 7496799 TI - Colocalization of glycine and GABA in synapses on spinomedullary neurons. AB - Spinomedullary neurons of the postsynaptic dorsal column pathway in adult cats were retrogradely labelled with horseradish peroxidase. Postembedding immunogold reactions were performed with antisera which recognise GABA or glycine to determine if synaptic boutons in contact with these neurons contain both transmitters. Analysis of series of ultrathin sections revealed that synaptic profiles with strong immunogold reactions for GABA usually also displayed strong immunogold reactions for glycine. Pre-embedding immunocytochemistry was performed on sections containing labelled cells with a monoclonal antibody which recognises the glycine receptor-associated protein, gephyrin. Many synapses onto postsynaptic dorsal column neurons were associated with gephyrin-like immunoreactivity and these typically contained irregularly shaped vesicles. Immunogold reactions showed that synaptic profiles apposed to gephyrin immunoreactive junctions contained GABA and glycine. The evidence suggests that glycine is a neurotransmitter at synapses on spinomedullary neurons and that it is colocalized with GABA. PMID- 7496800 TI - A light and electron microscopic study of calbindin D-28k immunoreactive double bouquet cells in the human temporal cortex. AB - Correlative light and electron microscopic methods were used to examine the morphology, distribution and synaptic connections of double bouquet cells immunoreactive for the calcium-binding protein calbindin D-28k in the human temporal neocortex. Double bouquet cells form symmetric synapses with small dendritic shafts and dendritic spines. The distribution and proportion of synapses found in the present work are very similar to those found in previous studies on the synaptic connectivity of double bouquet cells in the monkey cerebral cortex. Thus, double bouquet cells are probably involved in similar synaptic circuits in monkeys and humans. PMID- 7496802 TI - Central cardiovascular and behavioral effects of carboxy- and amino-terminal fragments of substance P in conscious rats. AB - The central cardiovascular and behavioral effects of carboxy- (SP 5-11, SP 6-11, SP 7-11, SP 8-11) and amino- (SP 1-7, SP 1-9) terminal substance P (SP) fragments were compared with those of SP 1-11 in conscious rats. In addition, the ability of these SP-fragments to induce desensitization of the central NK1 receptor was investigated. SP 1-11 (50 pmol) injected i.c.v. induced an increase in mean arterial blood pressure (MAP), heart rate (HR) and a typical behavioral response consisting of face washing (FW), hindquarter grooming (HQG) and wet-dog shakes (WDS). The cardiovascular and behavioral responses to equimolar doses of SP 5-11 and SP 6-11 were similar to those of SP 1-11, however, only SP 5-11 induced exactly the same behavioral pattern as SP 1-11. SP 6-11 was more potent in inducing FW and WDS than SP 1-11 or SP 5-11. The carboxy-terminal SP-fragments, SP 7-11 and SP 8-11, and the amino-terminal SP-fragments, SP 1-7, SP 1-9, did not elicit any significant cardiovascular or behavioral responses. Pretreatment with SP 1-11 reduced the cardiovascular and behavioral responses to subsequent injections of SP 1-11. Of all SP-fragments tested, only SP 5-11 was able to attenuate the cardiovascular and behavioral responses to SP 1-11. Our results demonstrate that SP 6-11 represents the shortest carboxy-terminal amino acid sequence, that after i.c.v. injection, elicits the same cardiovascular response as SP 1-11, but fails to desensitize the NK1 receptor. The carboxy-terminal fragment, SP 5-11, is the shortest amino acid sequence which produces the same pattern of central cardiovascular and behavioral responses as SP 1-11 and also retains the ability to desensitize the NK1 receptor like SP 1-11. PMID- 7496801 TI - Calcitonin gene-related peptide innervation of A2-catecholamine cells in the nucleus of the solitary tract of the rat. AB - By using double immunolabeling light and electron microscopic techniques, dense neuronal network of calcitonin-related peptide (CGRP) has been visualized in the nucleus of the solitary tract with complete overlapping of the tyrosine hydroxylase (TH)-containing cells. TH-immunoreactive perikarya and dendrites were seen in synaptic contact with CGRP-immunopositive fibers, indicating that CGRP, by carrying sensory signals may influence autonomic regulatory mechanisms in the NTS through local catecholaminergic neurons. PMID- 7496803 TI - Identification of endothelin receptor subtypes in rat retina using subtype selective ligands. AB - We investigate the existence of endothelin receptor subtypes using subtype selective ligands and the presence of immunoreactive (IR) endothelin (ET)-3 (IR ET-3) by radioimmunoassay (RIA) in rat retina. Scatchard transformation of saturation binding experiments with [125I]ET-3 revealed specific binding sites with a Kd and Bmax values of 42 +/- 12 pM and 111 +/- 24 fmol/mg of protein, respectively. The Kd was similar to that obtain in previous studies using [125I]ET-1. However, the Bmax was 65% of that obtained with [125I]ET-1. Competitive experiments in the presence of the cyclic pentapeptide BQ123 (selective for ETA receptor) and Sarafotoxin 6C (selective for ETB receptor), demonstrated the existence of ETA and ETB receptors in a ratio of 35:65. The order of potency of ET family peptides was ET-3 = ET-1 > S6C for ETB receptor and ET-1 > ET-3 > BQ123 for ETA receptor. Cross-linking of [125I]ET-1 to retinal membranes with disuccinimidyl suberate and SDS-PAGE followed by autoradiography resulted in the labeling of two bands with apparent molecular masses of 52 and 34 kDa. Similar results were obtained using [125I]ET-3, suggesting that ETA and ETB receptors have similar molecular mass. The 34 kDa band is a proteolytic degradation product of the 52 kDa band. The concentration of IR-ET-3 was 1212 +/- 153 fmol/g wet weight in rat retina. All these data suggest that ETs may play a role in neurotransmission or neuromodulation in the retina, operating on both ETA and ETB receptor subtypes present in this tissue. PMID- 7496804 TI - Effects of CGS 21680 in vivo on dopamine D2 agonist binding in the rat brain. AB - To investigate whether adenosine A2a agonists modulate dopamine D2 receptor binding in vivo, we have analyzed the effects of intraperitoneally administered 2 [p-(carboxyethyl)phenylethylamino]-5'-N-ethylcarboxamidoadenosin e (CGS 21680) on the ability of dopamine to compete at [125I]iodosulpride (0.25 nM) binding sites in filter-wiped cryostat sections of the rat forebrain and on [3H]L-(-)-N propylnorapomorphine ([3H]NPA) binding (1 nM) using quantitative receptor autoradiography. CGS 21680 (1-3 mg/kg) decreased the IC50 value of dopamine on [125I]iodosulpride binding, and the decrease at 1 mg/kg was blocked by the A2 antagonist 3,7-dimethyl-1-propargylxanthine (DMPX; 5 mg/kg). The decrease in the IC50 value of dopamine was due to a decrease in the KL value whereas the KH value and the proportion of high-affinity binding sites were unaffected. The binding of [3H]NPA was significantly increased in the rostral and caudal parts of the caudate-putamen and in the rostral part of the olfactory tubercle, whereas no change could be demonstrated in the nucleus accumbens and in the caudal part of the olfactory tubercle. These results indicate that stimulation of A2a receptors in vivo causes alterations in the binding characteristics of D2 receptors in certain regions of the basal ganglia. PMID- 7496805 TI - Down-regulation of the hypothalamo-pituitary-adrenal axis reduces brain damage and number of seizures following hypoxia/ischaemia in rats. AB - Several reports suggest that the activity of the hypothalamo-pituitary-adrenal axis (HPA-axis) is increased following hypoxia/ischaemia and that this might be associated with increased neuronal vulnerability. The main goal of this study was to examine the effects of down-regulation of the HPA-axis on the hypoxia/ischaemia-induced (1) rise of plasma corticosterone levels, (2) seizures, and (3) brain damage. Down-regulation of the HPA-axis was induced by prolonged corticosterone treatment lasting until 24 h before hypoxia/ischaemia exposure. When compared to 8 days vehicle (sesame oil)-treated animals (CONT), 8 days daily corticosterone (40 mg/animal)-treated animals (CORT) showed significantly reduced adrenal-and thymus weight. Shortly after hypoxia/ischaemia plasma corticosterone levels in CORT animals were significantly reduced (17.30 micrograms/dl +/- 3.50) when compared to CONT animals (54.80 micrograms/dl +/- 7.78). This correlated with the brain damage which is expressed as the ratio between the damaged area and the total area. The total brain damage was significantly less in CORT-treated animals (28% +/- 11%) than in CONT animals (69% +/- 2%). Following hypoxia/ischaemia the number of seizures was significantly reduced in CORT animals (56 +/- 26) when compared to CONT animals (217 +/- 50). We conclude that prolonged corticosterone treatment resulting in down-regulation of the HPA-axis leads to (1) lower plasma corticosterone levels during hypoxia/ischaemia, (2) a reduction in brain damage following hypoxia/ischaemia, and (3) less hypoxia/ischaemia-induced seizures. PMID- 7496806 TI - Induction of Fos protein in a model of closed head injury in rats. AB - The induction of Fos immunoreactivity in the pyriform cortex and the hippocampal formation after closed head injury was determined and compared to that seen following cortical needle injury in pentobarbital anaesthetized male rats. Robust Fos expression was observed in the ipsilateral pyriform cortex following both types of injury but was observed in the ipsilateral dentate gyrus only following closed head injury. Pretreatment with the NMDA receptor blocker MK-801 eliminated closed head injury induced Fos expression in the pyriform cortex and attenuated that seen in the hippocampus. Similar amounts of Fos expression were observed in urethane anaesthetized lactating and nonlactating rats following closed head injury. No gross behavioural impairments as reflected in body weight gain and locomotor activity were seen in animals subjected to closed head injury. These results demonstrate that as with other forms of brain damage, closed head injury at levels that produce no overt brain lesion nor gross behavioural impairment induce Fos expression in the pyriform cortex and the dentate gyrus which is dependent on the activation of NMDA receptors. Further, this response to brain injury is not modulated by lactation. PMID- 7496807 TI - Retrograde axonal transport of signal transduction proteins in rat sciatic nerve. AB - Neurons require a mechanism to transmit stable signals over the large distance from the nerve growth cone or terminal to the cell body, in order that information from the target tissue can be relayed to the cell body where it is required. Nerve growth factor (NGF), a target-derived neurotrophic factor, is thought to signal over this distance by receptor mediated internalization of NGF, followed by retrograde axonal transport of the NGF-receptor complex. In this paper we show, by immunohistochemistry of rat sciatic nerve, accumulation of phosphotyrosine immunoreactivity only on the distal side of a nerve crush, suggesting axonal transport of tyrosine kinases and/or tyrosine phosphorylated proteins primarily in a retrograde direction. Furthermore, we also show retrograde axonal transport of phosphoinositide 3-kinase, ERK, MEK and MEK kinase, of which all but MEK kinase are known to be activated downstream of tyrosine receptor kinase activation. The retrograde transport of these proteins suggests that they may be involved in transmission of signals along the axon, relaying neurotrophic factor receptor activation at the nerve terminal to the nerve cell body. PMID- 7496808 TI - Photoperiod affects the expression of neural cell adhesion molecule and polysialic acid in the hypothalamus of the Siberian hamster. AB - Immunohistochemical and immunoblot procedures were used to examine the effects of inhibitory short day (SD) photoperiod on the expression of neural cell adhesion molecule (NCAM) and its polysialylated form (PSA-NCAM) in the hypothalamus and preoptic area (POA) of the adult male Siberian hamster. In animals that had undergone SD-induced gonadal regression, immunoblot analyses revealed significant reductions in the content of immunoreactive PSA in anterior hypothalamic (AH) and mediobasal hypothalamic (MBH) regions. These changes were accompanied by increased contents of the 180 kDa NCAM isoform in the POA and AH, and decreased content in the MBH. The 140 kDa NCAM isoform also was elevated in the AH. Light microscopic analysis revealed a marked reduction in the density of NCAM immunoreactive tanycyte-like processes in the MBH of animals exposed to SD. This effect was not blocked by castration, indicating that this may be a primay (sex steroid-independent) effect of altered photoperiod in the hypothalamus. Also, photoperiod-induced alterations in NCAM expression were not evident in non responsive hamsters that maintained active testes under SD exposure. Collectively, these results are evidence that seasonal changes in photoperiod affect the expression of NCAM and PSA in the hypothalamus. Such changes could help promote plastic morphological rearrangements related to the regulation of seasonal reproductive and/or metabolic cycles. PMID- 7496809 TI - Rat B-50 gene transcription and translation. AB - Previously we reported that the rat B-50/GAP-43 gene contains two promoters (P1 and P2). This study describes the contribution of these two promoters to the mRNA population in several paradigms leading to an altered B-50 mRNA expression. In 8 day-old rat brain we found that P1 transcripts (1676 +/- 50 nt) account for 5% and P2 transcripts (1462 +/- 46 nt) for 95% of the B-50 mRNAs. The expression of P1 and P2 derived transcripts is high at postnatal day 8 and the ratio between the amount of transcripts derived from P1 and P2 did not change during (embryonal and postnatal) development or aging. After peripheral nerve crush or transection B-50 mRNA expression in induced in the distal nerve stump. The amount of transcript in the nerve stump distal of the lesion derived from both P1 and P2 was increased and the ratio between P1 and P2 transcripts was not altered. To determine whether both P1 and P2 transcripts are translated, a polyribosomal profile from 8-day-old rat brain was generated. Northern analysis showed that both transcripts were associated with approximately four ribosomes. Since no change could be found in the activity in either of the two promoters under the different circumstances tested, we conclude that the activity of the two rat B-50 gene promoters is regulated by a similar mechanism. PMID- 7496810 TI - Microglial responsiveness as a sensitive marker for trimethyltin (TMT) neurotoxicity. AB - Activation of microglia is a well-documented phenomenon associated with diverse pathological conditions of the central nervous system. In order to investigate the involvement of microglial cells in the neurotoxic action of the heavy metal compound trimethyltin, three-dimensional brain cell cultures were treated during an early developmental period, using concentrations at or below the limit of cytotoxicity. Microglial cells were studied by cytochemical staining, using horseradish peroxidase-conjugated B4 isolectin (GSI-B4). In parallel, neurotoxic effects were assessed by determining the content of synaptophysin and synapsin I, both in the total homogenates and in the synaptosomal fraction of the cultures. Changes in the content of the specific growth cone protein, GAP-43, were also analyzed. It was found that low, non-cytotoxic concentrations of TMT (10(-9) to 10(-8) M) caused a significant increase in the number and/or the clustering of microglial cells. A decrease in the synaptic protein (synapsin I, synaptophysin) content was detected at 10(-8) M of TMT in synaptosomal fractions, whereas in the total homogenates, changes in synaptic proteins and GAP-43 were observed only at the cytotoxic TMT concentration (10(-6) M). Although it remains to be shown whether the microglial response is caused by direct or indirect action of TMT, the present findings show that microglial responsiveness can be detected prior to any sign of neuronal degeneration, and may serve as a sensitive indicator for heavy metal neurotoxicity in the brain. PMID- 7496811 TI - The dynamic response of Golgi tendon organs to tetanic contraction of in-series motor units. AB - Golgi tendon organs (TOs) display a dynamic response related to the rate of tension development by the motor units (MUs) which activate each TO. When several MUs are activated together the response increases non-linearly with respect to tension. The dynamic response has been re-examined by recording the responses of TOs, in the adult cat, to tetanic, isometric contractions of one or more MUs and has been found to comprise two components: for each MU-TO pair the instantaneous frequency between the first two impulses of the response was linearly related to the rate of rise of the tension, the slope of the relationship being greatest for the slow units. Similarly, when activating MUs were stimulated together, the relationships between frequency and the initial rate of tension rise were linear. When the relationships were expressed in terms of relative tension rate the organs were found to be most strongly activated by the IIb muscle fibres, the responsiveness at the level of the whole muscle being reduced by unloading effects. The later component of the response showed an exponential relationship between discharge frequency and rate of tension development, the slope of the relationship between frequency and the logarithm of tension rate being independent of MU size. Although absolute tension increases during a contraction, this did not appear to impinge on the response which was dominated by the dynamic parameters of the contraction. PMID- 7496812 TI - Neuronatin mRNA: alternatively spliced forms of a novel brain-specific mammalian developmental gene. AB - Neurogenesis begins with the closure of the neural tube around mid gestation and continues in the rat for about two weeks postnatally. Therefore, we investigated the role of neuronatin, a novel cDNA that we cloned from neonatal rat brain (Joseph et al., Biochem. Biophys. Res. Commun., 201 (1994) 1227-1234), in brain development. Further studies described in the present manuscript, lead to the identification of two alternatively spliced forms of neuronatin mRNA, alpha and beta, with the same open reading frame. Neuronatin-alpha encoded a novel protein of 81 aa, and the beta-form encoded 54 aa. Both forms were identical, except that the alpha-form had an additional 81 bp sequence inserted into the middle of the coding region. On Northern analyses, neuronatin mRNA was relatively selective for the brain. It first appeared at E11-14, a time when the neural tube has closed and neuroepithelial proliferation initiated, became pronounced at E16-19 with a surge in neurogenesis, and declined postnatally to adult levels with the completion of neurogenesis. In order to determine whether there were other forms of neuronatin mRNA, and to study the expression of the alpha and beta forms separately during development, reverse transcriptase-polymerase chain reaction was carried out using primers flanking the coding region of the alpha and beta forms. The RT-PCR results clearly indicated that there were only two forms of neuronatin. The beta-form first appeared at E11-14, whereas the alpha-form was present even earlier at E7-10. Together, these findings indicate that the two forms of neuronatin mRNA are regulated differently during brain development.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7496813 TI - Effects of intrahypothalamic morphine and its interactions with oxytocin and vasopressin during food intake in rats. AB - The effects of intra-hypothalamic microinjections of morphine (MOR) into the ventromedial (VMH) and lateral (LH) nucleus and its interactions with oxytocin (OXY) and vasopressin (AVP) were evaluated during food intake FI) in rats. Acute MOR clearly enhanced FI during the dark phase (6-24 h) of the diurnal cycle, while no appreciable changes in the light phase (0-6 h) FI was seen after intra VMH or LH injections. Long term MOR administration into both VMH and LH attenuated the enhancements in FI seen after acute MOR during the dark phase, while FI was enhanced during the light phase. Pretreatment with OXY or AVP clearly antagonized such tolerance or reverse tolerance/sensitization development to the orexic effect of MOR. These results are discussed in the light of complex peptidergic interactions in the hypothalamus during ingestive behavior. PMID- 7496814 TI - Subfornical organ stimulation elicits drinking. AB - The subfornical organ is a specialized central nervous system structure known to be involved in the control of drinking. We report here that electrical activation of subfornical organ neurons (100 microA, 10 Hz, 1 ms pulse width, for 5 min.) induced drinking in 67% (six of nine) of animals. This effect was site-specific as none of the animals with electrode placements (n = 7) dorsal or rostral to the subfornical organ drank in response to electrical stimulation. In contrast, activity levels were increased significantly during stimulation in both the subfornical and nonsubfornical organ stimulated groups. These results suggest that electrical stimulation of the subfornical organ can elicit site-specific drinking behaviour and provide further support for the involvement of the subfornical organ in the control of drinking behaviour. PMID- 7496815 TI - Rat pineal cell aggregates: ultrastructural and functional characteristics. AB - The aggregates were obtained by constant gyratory shaking of suspension cells freshly isolated from adult rat pineal glands. Their sizes ranged from 60 to 120 microns. Within 4-5 days, the aggregates formed by pinealocytes, astrocytes, and other unidentified cells became organized in a tissue-like configuration. There was no proliferation of the fibroblast cells. Ultrastructural characteristics of the aggregates were revealed by the presence of granular lysosomes, which are typical of pinealocytes, and are actively involved in the secretion. Functional characteristics were studied in static incubation. The aggregates secreted melatonin and other indole amines in culture medium. Basal melatonin release was detected until Day 24 of culture. This secretion was stimulated 230% with Isoproterenol (beta-adrenergic agonist), 725% with Epinephrine (alpha- and beta adrenergic agonists), and 140% with Vasoactive Intestinal Peptide after 5 days in culture, then > 1200% with Forskolin 9 days later (14-day-old aggregates). The results indicate that three-dimensional aggregates obtained from isolated pineal gland cells were the functional multicellular structures with in vivo characteristics. PMID- 7496816 TI - Long-term effects of human-to-rat mesencephalic xenografts on rotational behavior, striatal dopamine receptor binding, and mRNA levels. AB - Fetal ventral mesencephalic grafts have been used as a tool to counteract the symptoms of Parkinson's disease. In this study human fetal ventral mesencephalic xenografts were implanted into the lateral ventricle of unilaterally dopamine depleted immunosuppressed rats. Rotational behavior elicited by low doses of apomorphine, host striatal dopamine receptor binding, and mRNA levels were investigated. Rotational behavior was reduced beginning 2 months after grafting. After 4 months only a small number of rotations, lasting approximately 30 min, were recorded. Seven months after transplantation, the rotational behavior was completely eleminated. Dopamine D2 receptor binding revealed significantly increased levels in sham-operated 6-hydroxydopamine- (6-OHDA) lesioned control striata. These increased levels decreased, and although still significantly higher at 4 months, normalized at the survival time of 7 months postgrafting. Regional differences were still obvious at 7 months in the dorsolateral quadrant of dorsal striatum. Dopamine D2 receptor mRNA revealed significantly increased levels in the lateral aspects of 6-OHDA-lesioned control striata, reversing by 4 months postgrafting. The D1 receptor binding revealed a moderately reduced signal in striata of lesioned animals. After grafting, this reduction became significantly lower than that seen in the control side, with a continous decrease over time. The same pattern was detected using in situ hybridization for dopamine D1 receptor mRNA, that is, moderate decreases after dopamine depletion and a significant decrease in the dorsomedial part of dorsal striatum 7 months postgrafting. Dopamine D3 receptor binding was increased after dopamine depletion, but reversed already by 4 months postgrafting. Taken together, human ventral mesencephalic xenografts are able to completely reverse apomorphine induced rotational behavior, provided the grafts are left in vivo for a sufficient time. The increased striatal D2 receptors are reversed after grafting, but the human xenograft further suppressed the D1 receptor subtype both at binding and at mRNA levels. There was no strict correlation in the time courses of dopamine receptor changes and reduction of rotational behavior. PMID- 7496817 TI - Oxidative enzyme activity in the rat soleus muscle and its motoneurons during postnatal maturation. AB - The effect of postnatal maturation on changes in the oxidative enzyme (succinate dehydrogenase) activity in the rat soleus muscle and its motoneurons was examined at 3, 6, and 12 weeks of age. The motoneurons innervating the soleus muscle were identified using the fluorescent retrograde neuronal tracer Nuclear Yellow. An inverse relationship between soma size and oxidative enzyme activity of soleus motoneurons was observed at 3 and 6 weeks of age, whereas there was no correlation between them at 12 weeks. Although the oxidative enzyme activity in the soleus muscle increased during postnatal maturation, it showed a decrease in the soleus motoneurons. These data demonstrate that the inverse relationship between soma size and oxidative enzyme activity of rat soleus motoneurons can only be detected in the early postnatal period and that the oxidative enzyme activity in the rat soleus muscle and its motoneurons can change independently during postnatal maturation. PMID- 7496818 TI - Gastric distension modulates hypothalamic neurons via a sympathetic afferent path through the mesencephalic periaqueductal gray. AB - The effects of gastric distension on extracellularly recorded single neuronal activity in the lateral hypothalamuslateral preoptic area-medial forebrain bundle (LPA-LH-MFB), other areas of the hypothalamus, mesencephalic periaqueductal gray (PAG), and other areas associated with the mesencephalic reticular formation were determined in the anesthetized rat. Gastric distension was produced by filling a gastric balloon with water using a calibrated infusion pump. Experimental conditions were based on previous studies that simulated gastric distension during fluid consumption in the rat. The effects of stomach distension using water at body temperature and room temperature were compared. Neurons in both the hypothalamus and mesencephalon were modulated by gastric distension. Hypothalamic neurons exhibited responses associated with gastric distension and exhibited interactions between distension and temperature stimulation of the stomach. Neurons in the mesencephalic periaqueductal gray (PAG) and associated reticular formation also were modulated by these gastric stimuli. When the PAG was electrically stimulated, similar responses to gastric distension and PAG stimulation were observed for hypothalamic neurons. The effects of gastric distension on hypothalamic neurons were reduced or eliminated when the PAG stimulating electrode site was destroyed by electrocoagulation. In addition, the microiontophoretic application of horseradish peroxidase at hypothalamic neuronal recording sites where gastric distension effects were observed resulted in the retrograde labeling of neurons in the PAG. These gastric stimulation-induced effects on hypothalamic and mesencephalic neuronal activity were attenuated but were not permanently eliminated by bilateral cervical vagotomy. However, these effects were significantly reduced or eliminated by bilateral transection of the cervical sympathetic chain or spinal transection at the first cervical level. Because the filling of balloons placed into the abdominal cavity close to the stomach had no similar effects on neural activity, these results can be attributed primarily to the activation of gastric mechano-and temperature sensitive receptors. These results indicate that the effects of gastric temperature/distension stimulation under these conditions are mediated to a large degree by sympathetic afferents. The PAG is clearly involved as one of the mesencephalic relays for gastric afferent input to the hypothalamus. PMID- 7496819 TI - Catecholamine and NPY efferents from the ventrolateral medulla to the amygdala in the rat. AB - Anatomical tracing studies have demonstrated an efferent pathway to central nucleus of the amygdala (CeA) from the ventrolateral medulla. The combined retrograde tracing/immunohistochemical method was used to test for the presence of catecholamines and neuropeptide Y (NPY) in ventrolateral medulla neurons that innervate the CeA. Numerous retrogradely labeled neurons were observed in the ventrolateral medulla caudal to the area postrema. Fewer retrogradely labeled neurons were also observed in the rostral ventrolateral medulla. Retrogradely labeled neurons immunoreactive for both tyrosine hydroxylase and NPY were found in the caudal ventrolateral medulla. Double-labeled phenylethanolamine-N methyltransferase neurons were present at levels rostral to the area postrema. A substantial portion of the ventrolateral medulla projection to the CeA arises from adrenergic cells of the C1 group, because nearly 40% of the retrogradely labeled cells were also immunoreactive for phenylethanolamine-N-methyl transferase. The high percentage of double-labeled NPY-immunoreactive neurons suggests NPY is colocalized in CeA-projecting catecholamine neurons, indicating that input from the ventrolateral medulla to the CeA primarily arises from C1 adrenergic neurons that also express NPY. This contrasts with previous data suggesting the catecholaminergic projection from the nucleus of the solitary tract to the CeA originates from the A2 noradrenergic cell group. Thus, the input to the CeA from catecholaminegic groups in the caudal medulla is chemically coded in terms of a dorsal noradrenergic and a ventrolateral adrenergic pathway. PMID- 7496820 TI - Convergence of laryngeal afferents with different natures upon cat NTS neurons. AB - To clarify the convergence of laryngeal afferents within the nucleus tractus solitarius (NTS) in the cat, we examined in the medulla the response characteristics of superior laryngeal nerve (SLN) fibers and NTS neurons to mechanical and chemical stimulation applied to laryngeal mucosa by extracellular recordings. The response was recorded in 75 SLN primary afferent fibers (PAFs) and 92 NTS neurons. PAFs of the SLN consisted of numerous monomodal mechanosensitive fibers and a small number of chemosensitive and polymodal fibers. On the other hand, the majority of NTS neurons had a polymodal nature. Thus, laryngeal information is considered to be integrated in modality into NTS neurons. All mechanosensitive fibers received information from a small restricted field in the larynx. On the contrary, each NTS neuron responded to mechanical stimulation over a wide laryngeal field, indicating that information from different sites spatially converge on NTS neurons. Our results suggest that caudal NTS neurons play a cardinal role in integrating laryngeal afferents, which are thought to elicit laryngeal reflexes. PMID- 7496821 TI - Central projections of primary afferents from the interosseous nerve in the pigeon. AB - The interosseous nerve in the pigeon's leg innervates a string of Herbst corpuscles. Because Herbst corpuscles are vibration-sensitive, this study, using neuronal tracing methods, was expected to show the central representation of vibration sense. After application of a mixture of free and lectin-conjugated horseradish peroxidase to the interosseous nerve, labeled cell bodies of sensory and postganglionic neurons were mainly located in the dorsal root ganglia and paravertebral sympathetic ganglia L3/L4. In spinal segments L3/L4 fibers and terminals were mainly distributed at the lateral border of the head of the dorsal horn. In more cranial or caudal segments terminal fields were at intermediate parts of laminae I/II and laminae IV/V. Some labeled fibers entered the dorsal horn from medial to terminate in lamina IV. Primary afferents of the interosseous nerve projected directly to the gracile nucleus in the brainstem and distributed all along its rostrocaudal extent. Because the main terminal fields in the spinal cord are typical for the projection of small afferent fibers, vibration information seems to reach the brainstem via the dorsal column primary afferents. PMID- 7496822 TI - Circadian rhythm generation in the cultured suprachiasmatic nucleus. AB - The suprachiasmatic nucleus (SCN) of the hypothalamus is involved in the generation and entrainment of circadian rhythms. The results of a series of experiments in long-term cultured organotypic SCN slices suggest that (1) some but not all SCN neurons display circadian rhythmicity in their extracellular discharges. To the extent they could be studied, these neurons did not usually show synchronized high/low levels of activity; (2) simultaneous daily depolarization of these neurons (K+ pulses) to some extent influenced the distribution of the firing rate of SCN neurons around the time of expected daily pulses; (3) extracellular Ca++ and synaptic input is required for the pacemaker activity of the SCN. We conclude that the mammalian biological clock is a heterogeneous neuronal system in which the circadian pacemaker rhythm is generated and entrained via complex interactions among SCN neurons. PMID- 7496823 TI - Muscle activation in the elbow-forearm complex during rapid elbow extension. AB - Normal human subjects performed extensions of the elbow from an initial position to a visually defined target as rapidly and accurately as possible at different angular amplitudes (9 degrees-54 degrees) and at a constant amplitude (36 degrees) under various additional instructions. Joint angle, angular acceleration, and electromyograms (EMGs) from agonist (triceps lateralis), antagonist (biceps), and antebrachial muscles were recorded. EMGs of the antagonistic muscles showed reciprocal activation of the first triceps, and then the biceps, which was followed by the second triceps activation and coactivation of both muscles. Movement amplitude changed the reciprocal activation, whereas the coactivation was unaltered. Additional instructions changed the coactivation in amplitude, initiation, and termination, whereas the reciprocal activation was relatively preserved. Under these conditions, EMGs of antebrachial muscles were always linked with the coactivation in amplitude and timing but not with the reciprocal activation. The linkage suggests that antebrachial muscle activation serves as an indicator of the coactivation. From the indicator, we infer that the coactivation initiates from the beginning of the rapid movements and changes during the dynamic movement phase. PMID- 7496824 TI - Unrestrained walking in cats with medial pontine reticular lesions. AB - The early postoperative effects of lesions, aimed to destroy the caudal pole of the nucleus reticularis pontis oralis (NRPO) and the rostral pole of the nucleus reticularis pontis caudalis (NRPC), were tested in freely moving cats, walking at moderate speed (0.4-1.0 m/s). In cats in which these structures were partly or completely destroyed, the main effect of lesions was an impairment of fore hindlimb coordination, as shown by a change in the relationships between the lateral and diagonal time shift durations and the step cycle duration. In the second week after the surgery the values of the slopes of linear regressions relating these variables were markedly changed as compared to the preoperative data. The results suggest that the NRPO and NRPC are involved in maintaining the proper forehindlimb coordination during unrestrained locomotion in cats. PMID- 7496825 TI - Reported sequence homology between Alzheimer amyloid770 and the MRC OX-2 antigen does not predict function. PMID- 7496826 TI - Electrocution in a bath. AB - A wife was found dead by her husband, lying in a tiled bath on a cast concrete floor, with a 1 kW electric radiator immersed in the bath water. Initially the case was treated as accidental death, but the police charged the husband with murder on the grounds that his wife could not have died in the manner he described. Subsequent investigation showed that the bath had a low resistance to earth and that there were faults in both the house wiring and the radiator cable, which, together with a high blood alcohol content, produced a fatal chain of events. PMID- 7496827 TI - The distribution of the HLA-DQ alpha alleles and genotypes in the Turkish population as determined by the use of DNA amplification and allele-specific oligonucleotides. AB - Allele and genotype frequencies for the HLA-DQ alpha locus were determined for use in forensic analyses in Turkey. The polymerase chain reaction and the reverse dot blot format were employed to detect six different HLA DQ alpha alleles, which were detected among 150 unrelated individuals with allele frequencies ranging from 7% to 26.7%. The distribution of the observed genotypes was in Hardy Weinberg equilibrium. The discrimination power of this system in the Turkish population sample was 0.94, and the allelic diversity was 0.81. This study provides support for the validity of the HLA-DQ alpha procedure for typing forensic samples. PMID- 7496828 TI - The relation between speech tempo, loudness, and fundamental frequency: an important issue in forensic speaker recognition. AB - In voice identification, the mean fundamental frequency is a powerful and by far the best known speaker-specific acoustic parameter. In some cases, however, large differences in fundamental frequency values in different speech samples from the same subject may occur. This observation gave rise to the hypothesis that this parameter may interact with others, particularly speech tempo and loudness, which may both attain extreme values under forensic circumstances. In an experimental investigation with a total of 50 subjects in two groups, no statistically significant relationship was found between speech tempo, loudness and voice fundamental frequency. Therefore no correction or normalising procedure need be applied prior to the assessment or interpretation of fundamental frequency means. The issue needs further investigation with spontaneous speech samples and material from casework. PMID- 7496829 TI - Head and neck surgery for the surgical oncologist. PMID- 7496830 TI - Cell adhesion in tumor invasion. PMID- 7496832 TI - Malignant potential in intestinal juvenile polyposis syndromes. AB - BACKGROUND: Unlike familial polyposis coli, where the premalignant nature of adenomatous polyps is well established, the cancer risk in juvenile polyposis has generally been considered not increased. METHODS: This study reviews all cases of juvenile polyposis reported in the English language to date to assess the occurrence and prognosis of carcinoma in the gastrointestinal tract. RESULTS: A total of 218 patients met the inclusion criteria. Mean age at diagnosis was 18.5 years (range: 9 months to 67 years). No gender preference was identified. The most common presenting symptom was chronic anemia, followed by acute gastrointestinal bleeding, rectal prolapse of polyp, protein-losing enteropathy, and intussusception. A family history of juvenile polyposis could be established in approximately 50% of patients, and associated congenital malformations were detected in 15%. Ninety-nine patients underwent 138 gastrointestinal operations: 121 colorectal, 12 gastric, and 5 small intestinal procedures. The development of a gastrointestinal carcinoma was reported in 36 cases (17%). Mean age at diagnosis of carcinoma was 35.5 years (range: 4-60 years). Most malignancies were located in the distal colon and rectum, with only one case of gastric and one case of duodenal carcinoma. Tumor stage at diagnosis was usually advanced, with poor survival figures. CONCLUSIONS: This study shows that juvenile polyposis syndromes carry a more significant risk of carcinoma than generally appreciated. Therefore, more intense endoscopic surveillance may be warranted, and definitive surgical options should often be considered in these syndromes. PMID- 7496831 TI - Role of the E-cadherin/alpha-catenin complex in modulating cell-cell and cell matrix adhesive properties of invasive colon carcinoma cells. AB - BACKGROUND: The clinical behavior of colorectal cancer depends on its ability to invade and metastasize. Metastatic cells must dissociate from other cells and invade through basement membrane and stroma. Cell-cell adhesion in epithelial cells is mediated by the cell surface protein E-cadherin in association with alpha- and beta-catenin, which link E-cadherin to the cytoskeleton. Decreased cell-cell adhesion and increased motility on laminin have been correlated with more poorly differentiated and aggressive carcinomas. METHODS: In this study, the RKO cell line, previously shown by us to lack E-cadherin expression, was transfected with the complementary DNA for E-cadherin. The transfectants were selected for high levels of surface expression by sequential FACS and examined in functional assays. RESULTS: In comparison to control transfectants, the E cadherin transfectants exhibited a more epithelial-like morphology, a 30% increase in Ca(2+)-dependent cell-cell aggregation, and a markedly reduced motility on the matrix proteins, collagen I and laminin. CONCLUSIONS: These data demonstrate that correction of a defect in the cadherin/catenin cell-cell adhesion complex, often found in poorly differentiated and highly invasive tumors, facilitates increased cell-cell adhesion and retards tumor cell migration on basement membrane and stromal proteins. PMID- 7496834 TI - Perineal wound closure with the rectus abdominis musculocutaneous flap after tumor ablation. AB - BACKGROUND: We describe the transpelvic rectus abdominis musculocutaneous flap for closing large, often irradiated, perineal defects in patients with cancer. METHODS: We studied 9 men and 17 women who underwent wide excision of advanced T3 and T4 tumors and who had preoperative chemotherapy, radiotherapy, or both. After tumor ablation, we used a transpelvic rectus abdominis musculocutaneous flap to fill pelvic dead space and primarily close the perineal wounds. Twelve simultaneous vaginal reconstructions were performed. RESULTS: Wounds healed promptly in 14 patients. Healing was delayed in the remaining 12 patients due to poor flap adherence to irradiated tissue beds (9), complete flap loss (2), or partial flap loss (1). Delayed healing was associated with the combination of obesity and prior irradiation. Other perioperative complications included localized abdominal wound infection (2), enterocutaneous fistula (1), and abdominal herniation (1). Local disease was controlled in 20 of 26 patients. Mean follow-up was 22 months. CONCLUSION: Transpelvic rotation of the rectus abdominis musculocutaneous flap facilitates healing large, difficult, perineal wounds, resulting from cancer ablation. PMID- 7496833 TI - Resection of brain metastases from sarcoma. AB - BACKGROUND: Brain metastases from sarcoma are rare, and data concerning the treatment and results of therapy are sparse. METHODS: We retrospectively reviewed 25 patients with brain metastases from sarcoma of skeletal or soft-tissue origin, surgically treated in a single institution during 20 years. RESULTS: In 18 patients the brain lesion was located supratentorially, and in 7 patients infratentorially. Median age at brain metastasis diagnosis was 25 years. Median time from primary diagnosis to diagnosis of brain metastasis was 26.7 months. Lung metastases were present in 19 patients and in 8 patients they were synchronous with the brain lesion. Pulmonary metastases were resected in 12 patients (48% of total, and 63% of those with pulmonary lesions). The overall median survival from diagnosis of the primary sarcoma was 38 months and from craniotomy was 7 months. The presence or absence of lung lesions did not alter the median survival as calculated from diagnosis of brain metastasis. Overall percent survival was 40% at 1 year and 16% at 2 years. CONCLUSIONS: Because brain metastases from sarcoma are refractory to alternative treatment, surgical excision is indicated when feasible. Brain metastases from sarcoma are uncommon, usually occurring with or after lung metastasis. Long-term survival is possible in some patients. PMID- 7496835 TI - Improved survival in young women with breast cancer. AB - BACKGROUND: Young age has been hypothesized to be an adverse prognostic factor for women with breast cancer. This association, based on historical data, may not reflect recent advances in breast cancer management. METHODS: A retrospective study was conducted of all women age 30 or younger who underwent definitive operation at our institution for primary operable breast carcinoma during one of two consecutive 20-year periods (1950-1969 or 1970-1989). All cancers were restaged according to current staging criteria. Actuarial survival and recurrence free survival rates from the two patient eras were compared with each other and with published statistics for older breast cancer patients. RESULTS: Eligibility criteria were met by 81 women from the 1950-1969 era and 146 women from the 1970 1989 era. Histologic diagnoses, tumor sizes, incidence of axillary nodal metastases, number of positive nodes, and American Joint Committee on Cancer stage at presentation were similarly distributed in the two eras. Despite these similarities, improved survival (p = 0.009) was observed in the later era. Local recurrences were also more common (p < 0.05) in the later era in association with less extensive resections. These local recurrences had an adverse impact on recurrence-free survival in the later era, but no concomitant decrease in overall survival was observed. Node-positive patients who received chemotherapy demonstrated a trend toward improved survival (p = 0.06) compared with node positive patients who did not. Survival for patients in the later era was similar to that for older women as reported in other published series. CONCLUSIONS: The stage of presentation of breast cancer in women 30 years or younger appears unchanged from prior decades, but survival has improved in association with the use of less extensive surgical resections and the introduction of cytotoxic chemotherapy. With current treatment, primary operable breast cancer in young women appears to have a similar prognosis to breast cancer in older women. PMID- 7496836 TI - Proliferative indices and oncoprotein expression in benign and malignant breast biopsies. AB - BACKGROUND: Prognostic factors are used routinely in the management of breast cancer. However, their potential for identifying precursor malignant lesions has not been assessed. METHODS: We have examined 285 breast biopsy specimens (140 benign, 145 malignant) for DNA ploidy, S-phase fraction, Ki-67 nuclear antigen proliferative indices, and HER-2/neu and epidermal growth factor receptor oncoproteins. RESULTS: When proliferative indices were compared between the benign and malignant groups, differences were noted for DNA ploidy, S-phase fraction, and cell cycling index (p < 0.0005). When the benign nonproliferative specimens were compared with the atypical/proliferative benign specimens, proliferative indices failed to show any differences. When the specific subset of proliferative/atypical benign breast tissue was compared with the malignant specimens, DNA index, S-phase fraction, and cell cycling index showed significant differences. The mean for epidermal growth factor receptor was greater in the non proliferative group but not statistically significant (p < 0.1). HER-2/neu oncoprotein failed to show any differences between the benign and malignant groups. Within the atypical benign group, Ki-67 correlated strongly with S-phase fraction and HER-2/neu (p < 0.01). CONCLUSIONS: We have demonstrated that proliferative indices can differentiate between benign and malignant breast tissues but not among specific subgroups. In addition, epidermal growth factor may differentiate between nonproliferative and proliferative/atypical benign biopsy results. Oncoprotein determination, ploidy, and DNA proliferative indices may be useful in defining malignant and benign breast disease but are not useful in distinguishing between benign and malignant breast disease with an increased likelihood for malignant transformation. PMID- 7496837 TI - Axillary node status in nonpalpable breast cancer. AB - BACKGROUND: The metastatic status of the axillary nodes is prognostically important but its value has been questioned in the management of nonpalpable breast tumors. This study correlates the incidence of positive nodes with the size of the primary nonpalpable tumors. METHODS: We retrospectively analyzed 220 invasive and 21 microinvasive breast cancers that were excised after needle localization and for which axillary dissections were subsequently performed. Of invasive cancers, 166 presented as mass lesions with or without microcalcifications and 54 as microcalcifications alone. The size of the mass lesions (n = 166) was determined mammographically and on pathologic specimens. They were subdivided into five groups according to diameter: (a) < or = 5 mm, (b) 6-10 mm, (c) 11-20 mm, (d) > 20 mm, and (e) unrecorded size. RESULTS: Axillary metastases were found in 9% of patients whose cancer presented as microcalcifications alone. They were found in 0, 11, and 22% of patients in mammographic groups, a, b, and c, respectively. In the corresponding groups in which size was determined from the pathology report, metastases were found in 5, 10, and 27%. CONCLUSION: The size of nonpalpable breast cancers measured on the excised gross specimen and by mammogram accurately predicts the likelihood of axillary node metastasis. PMID- 7496838 TI - Estrogen and progesterone receptors in thyroid lesions. AB - BACKGROUND: Given the higher incidence and better prognosis of thyroid cancers in women, the possibility arises that these lesions may be influenced by sex hormones. With the development of monoclonal antibodies to the estrogen and progesterone receptor proteins, receptor status can now be determined by immunohistochemical methods that allow direct localization of receptors in tissue. METHODS: Using this technique, we have studied tissues of 11 patients, 2 of them pregnant, with thyroid lesions. Paraffin-embedded tissues were used. Positive controls consisted of known estrogen- and progesterone-positive breast carcinomas. RESULTS: Examination of both the thyroid lesions and adjacent uninvolved thyroid tissue showed no nuclear reactivity with either estrogen or progesterone receptor antibodies. Our study did not confirm the previously reported incidence of estrogen and progesterone receptors in thyroid lesions. CONCLUSIONS: We conclude that contrary to earlier indications, estrogen and progesterone receptor proteins are neither significantly detectable nor pertinent for follow-up or prognosis in the patient with thyroid neoplasia. PMID- 7496839 TI - Modified instruments for mobilization of the ureters and parametrial transsection during radical hysterectomy. AB - BACKGROUND: The untunneling of the ureters and the transsection of the parametria are two well-characterized steps of radical hysterectomy. Two sets of instruments were developed to facilitate the mobilization (untunneling) of the ureters and the dissection of the paravaginal and parametrial tissues during radical hysterectomy. MATERIALS: Instrument set one consists of a modified Uchida spoon, two ureteral tunnel clamps, and a pair of tunnel scissors. Set two contains a long and a short sharply angled parametrial clamp with matching parametrial scissors. RESULTS: This technique permits one-step exposure of the ureter, protected from injury by the spoon. The instruments for parametrial transsection facilitate controlled transsection of the paravaginal and parametrial tissues while conserving surgical margins. They can also be conveniently used for transsection of the uterosacral ligaments. CONCLUSIONS: The authors believe that the instruments contribute to an enhanced safety and accuracy of radical hysterectomy with improved efficiency by shortened operating time. PMID- 7496840 TI - Prognostic variables in invasive breast cancer: contribution of comedo versus noncomedo in situ component. AB - BACKGROUND: Many invasive breast cancers are accompanied by a variety of noninvasive components. Histological distinctions have been made between these components, but to understand their importance, it is essential to examine their molecular biology. METHODS: Proliferative indices, oncoproteins, and steroid receptor expression were compared for invasive breast cancers containing comedo type ductal carcinoma in situ (n = 35), noncomedo-type ductal carcinoma in situ (n = 34), and pure invasive cancers (n = 49). Ploidy, S-phase fraction, Ki-67 staining, estrogen receptor (ER), progesterone receptor (PR), and the expression of HER-2/neu and epidermal growth factor receptor (EGFR) were evaluated in these tumors. RESULTS: The comedo-invasive subgroup differed significantly from the noncomedo-invasive subgroup, demonstrating significantly higher mean ploidy (1.6 vs. 1.3; p = 0.0156), S-phase fraction (7.9% vs. 4.3%; p = 0.0066), Ki-67 staining (20.3% vs. 12.0%; p = 0.0058), and HER-2/neu values (2,247 fm/mg vs. 1,014 fm/mg; p = 0.0412) and lower ER (76 fm/mg vs. 339 fm/mg; p = 0.006) and PR values (99 fm/mg vs. 265 fm/mg; p = 0.0608). A higher percentage of comedo invasive carcinomas demonstrated aneuploidy 71%; p = 0.0158), elevated levels of S-phase fraction (75%; p = 0.0016) and Ki-67 staining (55%; p = 0.0512), overexpression of HER-2/neu oncogene (47%; p = 0.0011), and were ER negative (35%; p = 0.0148), PR negative (47%; p = 0.0073) when compared to noncomedo invasive carcinomas. Comedo-invasive and noncomedo-invasive tumors were comparable for nodal status and tumor size, but differences were noted for tumor differentiation and percentage of tumors that were > 1 cm. Comedo-invasive tumors were predominantly poorly differentiated (60 vs. 32%) and were > 1 cm (94 vs. 77%, p < 0.05). RESULTS: Comedo-invasive cancers were comparable to pure invasive cancers for ploidy, S-phase fraction, Ki-67 staining, and ER, PR, and EGFR expression. However, comedo-invasive carcinomas had greater HER-2/neu overexpression when compared to pure invasive tumors (47 vs. 19%; p = 0.0359). CONCLUSIONS: These results are consistent with the hypothesis that comedo carcinoma is a more aggressive type of ductal carcinoma in situ and may have independent prognostic value when seen in association with infiltrating ductal carcinoma. In invasive tumors, comedo carcinomas are associated with poor prognostic factors, including higher ploidy, S-phase fractions, Ki-67 staining, negative ER and PR status, poorer differentiation, larger tumors, and presence of HER-2/neu oncogene overexpression. PMID- 7496841 TI - Adjuvant radiation therapy after axillary lymphadenectomy for metastatic melanoma: toxicity and local control. AB - BACKGROUND: Certain patients with locally advanced melanoma have a high risk of regional recurrence after surgical excision and lymphadenectomy alone. Growing evidence suggests that radiation therapy may improve local control with acceptable morbidity for patients with melanoma in some sites. There is no information regarding the safety or efficacy of this treatment when applied to the axilla for regional metastasis of cutaneous melanoma. METHODS: We conducted a retrospective evaluation of patients who received postoperative radiation therapy to the axillary lymphatics for malignant melanoma at the M. D. Anderson Cancer Center between 1980 and 1992. Twenty-eight patients were identified who had undergone a formal axillary lymph node dissection followed by postoperative irradiation to the full axilla and supraclavicular fossa. Twenty-two patients were irradiated using a hypofractionated treatment regimen (4-7 Gy/fraction), generally to 30 Gy in five fractions. RESULTS: Sixteen patients are currently alive. Actuarial survival at 5 years is 50.1%. Local tumor recurred or persisted in five patients, four of whom had clinically palpable tumor when they were irradiated. CONCLUSIONS: Postoperative irradiation of the axilla for malignant melanoma has acceptable toxicity, even in those with extensive axillary dissection. In this group, patients treated for subclinical disease had a low rate of recurrence (1 of 21, [5%]). PMID- 7496842 TI - Inhibition of bladder tumor cell implantation in cauterized urothelium, without inhibition of healing, by a fibronectin-related peptide (GRGDS). AB - BACKGROUND: Local recurrence after transurethral resection of bladder tumors (TURB) is common and might be diminished if free tumor cells within the bladder are prevented from reattaching. METHODS: In vitro inhibition of murine bladder tumor cells to an approximation of urothelial matrix with agents that might block attachment to components of the extracellular matrix, and in vivo inhibition of attachment in cautery-injured murine bladder. RESULTS: GRGDS, (0.1-2.5 mg/ml), a fibronectin-related peptide, mannose-6-phosphate, (0.1-20 mg/ml), a carbohydrate, and heparin (1-625 units/ml) all inhibited attachment in vitro in a dose dependent fashion. YIGSR (0.1-2 mg/ml), a laminin-related peptide, did not. Mannose (10 mg/ml) did not significantly inhibit attachment of tumor cells to cauterized urothelium in vivo, whereas there was a 77% reduction of attachment in bladders irrigated with GRGDS (6.25 mg/ml) (p < 0.05), and the appearance of subsequent tumors in the bladder was inhibited. Finally, GRGDS (6.25 mg/ml) did not inhibit healing of the cautery ulcer. CONCLUSIONS: RGD-containing peptides may be useful as adjuvant therapy to decrease local recurrence after TURB and perhaps in other circumstances in which tumor cells spilled into a wound or body cavity threaten surgical success. PMID- 7496843 TI - Autologous deposit of blood by oncology patients: an evaluation of actual use and wastage. AB - BACKGROUND: Transfusions of autologous deposited blood have continued to grow since the 1980s. Together with the growth in the deposit of autologous blood, issues such as appropriateness of indications, reimbursement, cost effectiveness, etc., have emerged and require addressing. Also, the patient's concerns about safety of the blood supply must be taken into account. METHODS: We conducted a 20 week study to determine the collection and transfusion/wastage of autologous blood and the need to transfuse additional allogeneic units of packed red blood cells. The disposition of every autologous unit was followed until its disposal. RESULTS: One hundred fifty-seven patients deposited 260 units of autologous blood. Of these, 144 units (55.4%) were transfused whereas 44.6% were discarded. In addition, 25 or 157 patients (15.9%) received an additional 58 units of allogeneic blood. CONCLUSION: These data demonstrate that there are surgical procedures where autologous blood is drawn unnecessarily whereas others could afford a higher number of autologous blood deposits. Physicians may be pressured by patients into ordering autologous blood deposits, to allay fear of patients about safety of the blood supply. It is important that health care professionals undergo periodic education to be able to address patient's apprehension about safety of the blood supply. PMID- 7496844 TI - Stomal seeding of head and neck cancer by percutaneous endoscopic gastrostomy (PEG) tube. PMID- 7496845 TI - Expected environmental conditions for the 1996 Summer Olympic Games in Atlanta. PMID- 7496846 TI - Fat-free mass index in users and nonusers of anabolic-androgenic steroids. AB - We calculated fat-free mass index (FFMI) in a sample of 157 male athletes, comprising 83 users of anabolic-androgenic steroids and 74 nonusers. FFMI is defined by the formula (fat-free body mass in kg) x (height in meters)-2. We then added a slight correction of 6.3 x (1.80 m - height) to normalize these values to the height of a 1.8-m man. The normalized FFMI values of athletes who had not used steroids extended up to a well-defined limit of 25.0. Similarly, a sample of 20 Mr. America winners from the presteroid era (1939-1959), for whom we estimated the normalized FFMI, had a mean FFMI of 25.4. By contrast, the FFMI of many of the steroid users in our sample easily exceeded 25.0, and that of some even exceeded 30. Thus, although these findings must be regarded as preliminary, it appears that FFMI may represent a useful initial measure to screen for possible steroid abuse, especially in athletic, medical, or forensic situations in which individuals may attempt to deny such behavior. PMID- 7496848 TI - The national jockey injury study: an analysis of injuries to professional horse racing jockeys. AB - While there have been numerous reports in the literature of accidents in equestrian sports, no comprehensive study has been conducted to ascertain the nature and incidence of injuries incurred by professional horse-racing jockeys. A survey was conducted to determine the types of injuries to jockeys and racing related health concerns, including weight reduction methods. The questionnaire was completed by 706 professional jockeys actively competing at United States racetracks between July and October 1990 about injuries they had sustained in their careers. More than 1,700 injuries were reported. Fractures (n = 1,113) accounted for 64% of the total. The most common cause of injury was becoming unseated, followed by the horse falling. Relationships between characteristics of jockeys and injuries were evaluated. There were significant numbers of serious injuries with prolonged periods of not riding. Recommendations for improving jockey safety are made. PMID- 7496847 TI - Risk factors for stress fractures in female track-and-field athletes: a retrospective analysis. AB - The incidence and nature of stress fractures and the relationship of potential risk factors to stress-fracture history were investigated retrospectively in a group of 53 female competitive track-and-field athletes. Forty-five stress fractures, diagnosed by clinical findings and bone scan, radiograph, or CT scan, were reported in 22 women. Tibial fractures were the most common (33%). There was no significant difference in bone mineral density at the lumbar spine and tibia/fibula or in percentage body fat and total lean mass when comparing the groups with and without a stress-fracture history. Athletes with a past stress fracture were significantly older at menarche and were more likely to have experienced a history of menstrual disturbance (p < 0.05). Analysis of dietary behavior found that athletes with stress fractures scored significantly higher on the EAT-40 test and were more likely to engage in restrictive eating patterns and dieting. Multiple logistic regression showed that athletes with a history of oligomenorrhea were six times more likely to have sustained a stress fracture in the past, while those who were careful about their weight were eight times more likely. Prevention and treatment of stress fractures in female athletes should include a thorough assessment of menstrual characteristics and dietary patterns. PMID- 7496849 TI - Situational correlates of emotional adjustment to athletic injury. AB - Patients at a sports medicine clinic served as subjects in an investigation of the relationship between various situational variables and emotional adjustment to athletic injury. Physician-rated current injury status, perceived impairment of sport performance, and perceived social support for rehabilitation were significantly correlated with postinjury depression. Age was negatively associated with postinjury mood disturbance. In general, patients demonstrated a positive mental health profile. The results suggest that although most patients cope well with their injuries, some patients experience clinically meaningful psychological distress that is associated with certain situational factors. PMID- 7496850 TI - A physician survey of therapy for exercise-associated amenorrhea: a brief report. AB - Amenorrheic athletes face an increased risk of osteopenia and stress fractures. Optimal treatment for exercise-associated amenorrhea remains controversial, reflecting limited data on the therapeutic effects of hormonal or nutritional intervention in the prevention of osteopenia. To determine physician opinions regarding preferred management of amenorrheic athletes, members of the American Medical Society for Sports Medicine (AMSSM) were surveyed by questionnaire. Practitioners were asked if they prescribed sex steroid replacement, calcium supplementation, weight gain, or decreased physical activity for amenorrheic athletes. The 159 respondents included predominantly sports medicine (56%) and family medicine (32%) physicians. Sex steroid replacement was endorsed by 92%, calcium supplementation by 87%, increased caloric intake by 64%, decreased exercise intensity by 57%, weight gain by 43%, and vitamin supplementation by 26%. These findings suggest that sex steroids are used commonly to treat amenorrheic athletes, despite the paucity of data demonstrating their efficacy in preserving bone mass in this disorder. Further research is needed to define the benefits of estrogen alone or in combination with nutritional intervention for preserving bone mass in female athletes. PMID- 7496851 TI - Occipital neuralgia in a football player: a case report. AB - Occipital neuralgia, a painful condition thought to be caused by nerve entrapment, has been described in the general population. Athletes, particularly those who participate in collision sports, such as football, may be predisposed to this condition. An illustrative case is presented along with discussion of the differential diagnosis and treatment of this type of headache. PMID- 7496852 TI - Two consecutive rib stress fractures in a female competitive swimmer. AB - We present a case of two consecutive stress fractures in a female swimmer. The diagnosis of the present stress fracture of the ninth rib was based on clinical history and examination and on a new fracture line and callus formation seen in consequent conventional radiographs. Based on the clinical history and radiography, the patient had suffered another rib stress fracture in the fifth rib 15 months earlier. No external trauma had preceded either of the fractures, and no secondary cause of stress fracture was found. Her anatomical and biomechanical characteristics and training errors seem to have been responsible for the stress fractures. Sports physicians should be aware of rib stress fractures. With prompt diagnosis the rest period is short. PMID- 7496853 TI - Noncontact knee dislocation in a female basketball player: a case report. AB - Knee ligamentous injuries occur during sport, and when there is extensive injury, they can be associated with subluxation or dislocation. We present the case of a female basketball player who sustained a knee dislocation during noncontact play. An immediate angiogram confirmed vascular integrity, and surgical treatment with ligamentous reconstruction was performed. Peroneal nerve injury was present but resolved in 42 weeks. A review of the literature discussing knee dislocation is presented. PMID- 7496854 TI - Ulnar stress fracture of the nondominant arm in a tennis player using a two handed backhand. AB - Stress fractures are infrequently seen in non-weight-bearing bones. However, stress fractures may be present in any bone that undergoes repetitive stress. We report an unusual case of stress fracture in the nondominant arm of a tennis player who uses a two-handed backhand stroke. PMID- 7496855 TI - Exercise during pregnancy and pregnancy outcome. PMID- 7496856 TI - Epidemiology of gastrointestinal complications of diabetes mellitus. AB - Diabetes mellitus is a common condition that affects multiple organ systems, including the gastrointestinal tract. Although several clinic-based studies have shown gastrointestinal symptoms to be frequent in diabetic patients, population based data are sparse. The impact of gastrointestinal complications on a typical person with diabetes in the community is largely unknown. PMID- 7496857 TI - Gastroparesis and dyspepsia in patients with diabetes mellitus. AB - About one-half of patients with insulin- or non-insulin-dependent diabetes have delayed gastric emptying (diabetic gastroparesis). Some of them complain of epigastric pain, nausea, vomiting or postprandial fullness (diabetic dyspepsia), although only a minority are severely symptomatic. Diabetic gastroparesis is clinically relevant not only by virtue of the symptoms induced but also because it may contribute to inadequate glycaemic control and impaired absorption of orally administered drugs. Recent data suggest that abnormal blood glucose control, not only autonomic neuropathy, contribute to the pathogenesis of disordered gastric motility. In most cases diabetic gastroparesis is diagnosed clinically in the absence of demonstrable lesions of the upper gastrointestinal tract. To evaluate gastric emptying, scintigraphy is the 'gold standard'. Gastrokinetic drugs are of help in the treatment of gastroparesis: erythromycin is the first choice in acute presentations and cisapride for chronic symptoms. New macrolides with prokinetic action and devoid of antibacterial properties are very promising and should add another pharmacologic approach to control dyspepsia and gastroparesis in diabetic patients in the future. PMID- 7496858 TI - Mechanism of postprandial hyperglycaemia in diabetes mellitus. AB - Abnormalities of hepatic and extrahepatic tissues combine to produce the postprandial hyperglycaemia characteristic of non-insulin-dependent diabetes mellitus. The altered metabolic responses are primarily due to defects in insulin secretion and action. Other regulators of carbohydrate metabolism also contribute to the altered milieu. PMID- 7496859 TI - Diarrhoea in patients with diabetes mellitus. AB - Diarrhoea in patients with diabetes mellitus may be due to anorectal or rectal dysfunction that results in incontinence, intestinal secretion or rapid intestinal transit, or may be associated with disorders that typically cause malabsorption. The latter include small bowel bacterial overgrowth, coeliac sprue and pancreatic insufficiency. A practical algorithm for diagnosis and advances in therapy is discussed. PMID- 7496860 TI - Incontinence and anorectal dysfunction in patients with diabetes mellitus. AB - The elucidation of the pathogenesis of faecal incontinence in patients with advanced diabetes mellitus has progressed during the last 15 years. In contrast to earlier concepts which promoted the concept that high stool volumes overwhelm normal continence mechanisms, recent studies indicate that the vast majority of diabetic patients with faecal incontinence have normal or only moderately increased daily stool volumes, but also exhibit multiple abnormalities of anorectal sensory and motor functions. These changes are not observed in continent diabetic patients. Treatment consists of pharmacologic and dietary interventions to modulate diarrhoea, and biofeedback techniques to improve rectal sensory thresholds and striated muscle responsiveness of continence mechanisms. This dual approach is often successful and is free of risks. PMID- 7496861 TI - Exocrine pancreatic function in diabetes mellitus. AB - The exocrine pancreas and pancreatic islet system have a multitude of complex anatomical and functional interrelations. In insulin-dependent diabetes mellitus, both exocrine pancreatic function and morphology appear to be altered, although this phenomenon and its responsible mechanisms have not been elucidated in detail in humans. However, reduced pancreatic secretion in insulin-dependent diabetes is not likely to result in impaired intraluminal nutrient digestion. Whether, the changes in exocrine pancreatic function and morphology are a result of disturbed islet (beta-cell) function or are due to a common immunological pathogenesis, as suggested by recent studies, remains to be clarified. PMID- 7496862 TI - Acetaminophen-induced hepatotoxicity: protection by enzyme inhibition. PMID- 7496863 TI - Ascites formation and dynamics in cirrhotic patients. PMID- 7496864 TI - The role of Helicobacter pylori eradication in the management of acute bleeding peptic ulcer. PMID- 7496866 TI - Ascites free-water dynamics in decompensated cirrhosis: effects of acute and chronic frusemide treatment. AB - OBJECTIVE: To evaluate the effects of acute (60 mg intravenous bolus) and protracted (75 mg per day orally for 1 week) frusemide administration on ascites volume, free-water peritoneal clearance (FWPC), intra-abdominal pressure (IAP) and body weight in 30 decompensated cirrhotic patients. METHODS: Individual ascites volume and FWPC were estimated by means of a previously validated methylene-blue dilution test. RESULTS: All the investigated parameters showed a significant reduction after the protracted diuretic treatment (ascites volume 9.0 +/- 1.0 to 4.9 +/- 0.61, P < 0.001; FWPC 99.8 +/- 6.7 to 76.2 +/- 6.6 ml/min, P < 0.001; IAP 17.0 +/- 1.5 to 11.3 +/- 0.8 cmH2O, P < 0.001; body weight 75.2 +/- 1.9 to 66.3 +/- 1.8 kg, P < 0.001). A significant FWPC reduction (89.5 +/- 14.6 to 68.8 +/- 12.1 ml/min, P < 0.02) was also observed after the acute frusemide administration in the absence of any appreciable modifications to ascites volumes and IAP. CONCLUSION: Frusemide treatment appears to be able to exert a direct significant effect on FWPC, which does not appear to be mediated by ascites volume or IAP modifications. PMID- 7496865 TI - Ketoconazole inhibits acetaminophen-induced hepatotoxicity in mice. AB - OBJECTIVE: To investigate the effect of ketoconazole on acetaminophen (AAP) induced hepatotoxicity in mice. MATERIALS AND METHODS: Mice were given AAP intragastrically (300 mg/kg) and treated with ketoconazole (100 mg/kg intraperitoneally) or saline either 30 min before or 2-3 h after AAP administration. Mortality was recorded for 48 h, during which all mice given saline either died or recovered fully. Serum alanine and aspartate transaminase levels were determined 24 h after administration of AAP. Prostaglandin E2, thromboxane A2 and leukotriene C4 production was determined 6 h after AAP administration in the supernatants from the short-term culture of liver fragments by radioimmunoassay. RESULTS: Ketoconazole significantly decreased mortality and transaminase levels when given to mice either 30 min before or 2 h after AAP. Liver fragments from mice with AAP hepatitis produced greater quantities of prostaglandin E2, thromboxane A2 and leukotriene C4 than fragments from normal liver. Pretreatment of mice with ketoconazole or its addition to liver fragments ex vivo further increased the production of prostaglandin E2 and reduced the production of thromboxane A2. The effect of ketoconazole on leukotriene C4 synthesis was different in vivo (synthesis stimulation) from in vitro (synthesis inhibition). CONCLUSION: The protective effect of ketoconazole in AAP hepatitis is most probably mediated by modulation of eicosanoid synthesis by liver cells. PMID- 7496867 TI - Helicobacter pylori and acute bleeding peptic ulcer. AB - OBJECTIVE: The prevalence of Helicobacter pylori in chronic peptic ulcer is well known. In this study the frequency of H. pylori infection was investigated in patients with acute bleeding peptic ulcer. DESIGN AND SETTING: Prospective study in a district hospital. PATIENTS: Seventy consecutive patients with acute bleeding peptic ulcer. INTERVENTIONS: Diagnosis was verified on admission by endoscopy, and healing was examined at follow-up. Previous history of ulcer disease, presence of dyspeptic symptoms and consumption of non-steroidal anti inflammatory drugs were recorded. MAIN OUTCOME MEASURES: H. pylori infection was detected by two serological tests in samples obtained on admission for the acute bleeding episode, and at follow-up 1-3 months later. RESULTS: With a commercial latex immunoassay, 53% of the patients with gastric ulcer and 62% with duodenal ulcer were shown to possess H. pylori antibodies. In the other test, a standard enzyme-linked immunosorbent assay based on cell surface protein antigens of H. pylori with high sensitivity and specificity, 81% of gastric ulcer patients and 85% of duodenal ulcer patients were shown to have H. pylori antibodies. CONCLUSION: The results indicate that H. pylori infection plays a major aetiological role in patients with acute bleeding peptic ulcer. PMID- 7496868 TI - Evidence against an increased risk of Helicobacter pylori infection in dentists: a serological and salivary study. AB - BACKGROUND: An oral-to-oral route of transmission of Helicobacter pylori infection has been postulated, which is supported by the observation that H. pylori is present in the saliva and in dental plaque. On the basis of this assumption, an increased risk of H. pylori infection among dentists was postulated. METHOD: Serum and salivary H. pylori immunoglobulin (lg)G antibodies were measured in a group of practising dentists. For comparison we also studied a group of controls from the same urban area matched for age, sex, smoking habits, alcohol and non-steroidal anti-inflammatory drug consumption, and history of dyspepsia. RESULTS: There was no significant difference in serum H. pylori lgG antibodies titres between dentists and controls [optical density (OD) 0.991 +/- 0.588 versus 1.025 +/- 0.591, respectively]. Salivary H. pylori lgG were 0.693 +/ 0.726 and 0.661 +/- 0.614 OD in the dentists and control groups, respectively. The frequency of H. pylori-seropositive subjects did not differ between the two groups [22 out of 39 (56%) versus 46 out of 71 (64%)]. A positive saliva assay was found in 23 out of 39 (59%) dentists and in 44 out of 71 (62%) controls. The odds ratio for a dentist being H. pylori-positive was 0.7 (95% confidence interval 0.3-1.7) by serology and 0.9 (95% confidence interval 0.4-2.1) by salivary antibody assay. CONCLUSION: The data of this study do not support the concept that dentists are a high-risk group for H. pylori infection. PMID- 7496869 TI - Monoclonal antibody 1F10 immunoreactivity in inflammatory bowel disease: a new marker specific only for continuous endothelial cells. AB - OBJECTIVE: To investigate in detail the immunohistochemical properties of the two endothelial-specific markers 1F10 (continuous endothelia) and MS-1 (discontinuous endothelia) in bowel tissues of patients suffering from chronic inflammatory bowel disease (IBD). METHOD: Immunohistochemical techniques were employed to study the morphology and phenotypic expression of these two proteins in routinely processed bowel tissues from 27 patients with Crohn's disease, 18 patients with ulcerative colitis, and 20 normal controls. RESULTS: All patients with IBD and controls showed a low to moderate 1F10 immunohistochemical staining restricted to the lamina propria and submucosa. In contrast to ulcerative colitis patients and healthy controls, 1F10 immunoreactivity was strongly upregulated in the muscularis propria of the small and large bowel in Crohn's disease patients regardless of the histological severity of the inflammatory process. We did not observe immunoreactivity for MS-1 on endothelial surfaces in either Crohn's disease or ulcerative colitis. CONCLUSIONS: We conclude that endothelia in patients with IBD do not undergo metaplasia. The high immunoreactivity of 1F10 antigen in the muscularis propria in Crohn's disease indicates a state of tropical immunological activation and may be important in the maintenance of chronic inflammation by facilitating leukocyte migration into sites of Crohn's disease involvement. Further studies of the factors controlling endothelial cell differentiation in the bowel of Crohn's disease patients may help to explain the features observed in this study. PMID- 7496870 TI - Antigen specificity of circulating anti-neutrophil cytoplasmic antibodies in inflammatory bowel disease. AB - OBJECTIVES: To characterize the antigen specificity of circulating anti neutrophil cytoplasmic antibodies (ANCAs) in inflammatory bowel disease (IBD). DESIGN: Analysis of the prevalence of circulating ANCAs in patients with ulcerative colitis and Crohn's disease, by both non-specific methods (immunofluorescence against fixed neutrophil leukocytes) and specific antigen techniques (against purified neutrophil leukocyte constituents). METHODS: Indirect immunofluorescence against fixed polymorphonuclear leukocytes, and solid phase enzyme-linked immunosorbent assay (ELISA) against neutrophil constituents (alpha-granules, elastase, myeloperoxidase, cathepsin g, lysozyme and lactoferrin). RESULTS: Although results using immunofluorescence were typical of other studies (ulcerative colitis positive in 41%, Crohn's disease in 10%), ELISA studies showed antibody activity against neutrophil components in 69% of patients with ulcerative colitis and 39% of those with Crohn's disease. Antibodies in ulcerative colitis were commonly directed (in descending order) against lysozyme, cathepsin G, elastase, and lactoferrin, and in Crohn's disease against lysozyme. CONCLUSION: Correlation of indirect immunofluorescence data and ELISA results indicated that even this large panel of specific antigens fails to identify all the ANCA targets in IBD. The lack of correlation between the findings of ANCAs, either in general or versus a specific target, and disease extent or activity in ulcerative colitis supports the suggestion that ANCAs are unlikely to be of primary importance in pathogenesis. PMID- 7496871 TI - Modulation of cytokine release from human monocytes by drugs used in the therapy of inflammatory bowel diseases. AB - BACKGROUND: Cytokines produced in the gut mucosa play an important role in the pathogenesis of inflammatory bowel diseases (IBD). To determine whether drugs used in the treatment of these diseases modulate cytokine synthesis, we investigated their effects on endotoxin-induced tumour necrosis factor (TNF) alpha, interleukin (IL)-1 beta and IL-6 release by elutriation-purified human monocytes in vitro. METHODS: Drugs tested were dexamethasone, 5-aminosalicylic acid, sulphapyridine and zileuton (a 5-lipoxygenase inhibitor). Monocytes were isolated and stimulated with endotoxin, and TNF, IL-1 and IL-6 levels were determined using an enzyme-linked immunosorbent assay. RESULTS: Monocyte stimulation with endotoxin resulted in an average TNF release of 2464 +/- 64 pg/10(6) cells, IL-1 release of 616 +/- 47 pg/10(6) cells and IL-6 release of 2259 +/- 148 pg/10(6) cells. Addition of dexamethasone resulted in a reduction of TNF, IL-1 and IL-6 release to below background levels. Sulphapyridine significantly reduced TNF and induced IL-1 release in a dose-dependent fashion, but had no significant effect on IL-6 release. 5-Aminosalicylic acid did not modulate IL-6 synthesis, but significantly reduced IL-1 and enhanced TNF synthesis. Zileuton reduced TNF and IL-6 release, but enhanced IL-1 release. CONCLUSION: We conclude that these anti-inflammatory drugs are able to modulate cytokine release by human monocytes. Further studies are needed to determine whether these effects are related to their therapeutic efficacy in IBD. PMID- 7496872 TI - Effects of erythromycin on gastric emptying, duodeno-caecal transit time, gastric and biliopancreatic secretion during continuous gastric infusion of a liquid diet in healthy volunteers. AB - OBJECTIVE: Erythromycin, a macrolide antibiotic, has been reported to increase gastric emptying. The aim of this study was to evaluate the effects of intravenous erythromycin (150 mg/h) on gastric emptying, small intestinal transit time, gastric and biliopancreatic secretions during gastric infusion of a liquid diet in healthy volunteers. DESIGN: A randomized double-blind crossover study (erythromycin versus placebo). METHODS: Gastric emptying rates of nutrients, gastric acid secretion, gastric pH, jejunal flow rates, as well as biliopancreatic secretions and duodeno-caecal transit time, were evaluated during a continuous infusion at 4.5 kcal/min of a nutrient solution (1 kcal/ml) in the antrum, over a 6 h period, by a perfusion method. RESULTS: During the 6 h period, total gastric volume and gastric acid secretion decreased during erythromycin administration of 37 and 22%, respectively (area under the curves). Lipase outputs were significantly higher with erythromycin than placebo. Bile salt output was not significantly different between erythromycin and placebo. Duodeno caecal transit time increased significantly during erythromycin infusion compared with placebo (191 +/- 12 versus 159 +/- 17 min; P < 0.05). CONCLUSION: During continuous gastric infusion of a liquid diet, intravenous erythromycin has a powerful effect on gastrointestinal function. The motor and secretory effects may enhance the tolerance and the efficiency of enteral nutrition in humans. PMID- 7496873 TI - Undefined malabsorption syndrome with villous atrophy successfully reversed by treatment with cyclosporine. AB - A 47-year-old man with a flat jejunal mucosa complicated by malabsorption, diarrhoea and lymphocytic colitis is presented. There was no response to gluten withdrawal alone, combination of a gluten-free diet and prednisone therapy, or total parenteral nutrition. Complete clinical remission was only achieved after simultaneous treatment with cyclosporine and a gluten-free diet. Rechallenge with a gluten-containing diet while cyclosporine treatment continued resulted in a relapse of diarrhoea and malabsorption. We conclude that cyclosporine may be an effective agent for the treatment of undefined, refractory forms of malabsorption. PMID- 7496874 TI - A case of systemic strongyloidiasis in an ex-coal miner with idiopathic colitis. AB - An ex-coal miner was diagnosed with an idiopathic sigmoiditis compatible with colitis ulcerosa. He was treated with corticosteroids because of his deteriorating clinical condition, but his condition continued to worsen. He revealed to have a systemic Strongyloides stercoralis hyperinfection and Strongyloides colitis. A short time after diagnosis he died from septic shock despite therapy with thiabendazole and antibiotics. S. stercoralis is an opportunistic infection that is not uncommon and has high mortality in hyperinfection. It is prevalent in (sub)tropical areas, and also in coal mines because of the specific microclimate. However, the parasite is difficult to detect because eosinophilia can appear normal in chronic infection. Several stool examinations, duodenal aspiration or biopsies are necessary for a high diagnostic sensitivity. Serological diagnosis is the most sensitive and specific but is not always available. PMID- 7496875 TI - Case reports of death associated with pseudomembranous colitis and clarithromycin. PMID- 7496876 TI - Internet as clinical information system: application development using the World Wide Web. AB - Clinical computing application development at Columbia-Presbyterian Medical Center has been limited by the lack of a flexible programming environment that supports multiple client user platforms. The World Wide Web offers a potential solution, with its multifunction servers, multiplatform clients, and use of standard protocols for displaying information. The authors are now using the Web, coupled with their own local clinical data server and vocabulary server, to carry out rapid prototype development of clinical information systems. They have developed one such prototype system that can be run on most popular computing platforms from anywhere on the Internet. The Web paradigm allows easy integration of clinical information with other local and Internet-based information sources. The Web also simplifies many aspects of application design; for example, it includes facilities for the use of encryption to meet the authors' security and confidentiality requirements. The prototype currently runs on only the Web server in the Department of Medical Informatics at Columbia University, but it could be run on other Web servers that access the authors' clinical data and vocabulary servers. It could also be adapted to access clinical information from other systems with similar server capabilities. This approach may be adaptable for use in developing institution-independent standards for data and application sharing. PMID- 7496877 TI - Information technology and medical education. PMID- 7496878 TI - Educational software evaluation process. AB - The Active Digital Library at the Vanderbilt University Medical Center has created and implemented an educational software evaluation process to facilitate the timely recommendation for product acquisition. Using this process, breadth and depth of subject coverage, clarity of presentation, quality of construction, and ease of use are being assessed by content and technical experts. The process uses a team approach, employing a bi-level evaluation instrument based on existing software evaluation forms and system bug reports. PMID- 7496879 TI - Evaluation of long-term maintenance of a large medical knowledge base. AB - OBJECTIVE: Evaluate the effects of long-term maintenance activities on existing portions of a large internal medicine knowledge base. DESIGN: Five physicians who were not among the original developers of the knowledge base independently updated a total of 15 QMR disease profiles; each updated submission was modified by a review of group serving as the "gold standard, " and the pre- and post-study versions of each updated disease profile were compared. MEASUREMENTS: Numbers and types of changes, defined as any difference between the original version and the final version of a disease profile; reason for each change; and bibliographic references cited by the physicians as supporting evidence. RESULTS: A total of 16% of all entries were modified by the updating process; up to 95% of the entries in a disease profile were affected. The two most common modifications were changes to the frequency of an entry, and creation of a new entry. Laboratory findings were affected much more often than were history, symptom, or physical exam findings. The dominant reason for changes was appearance of new evidence in the medical literature. The literature cited ranged from 1944 to the present. CONCLUSIONS: This study provides an evaluation of the rate of change within the QMR medical knowledge base due to long-term maintenance. The results show that this is a demanding activity that may profoundly affect certain portions of a knowledge base, and that different types of knowledge (e.g., simple laboratory vs expensive or invasive laboratory findings) are affected by the process in different ways. PMID- 7496881 TI - Computerizing guidelines to improve care and patient outcomes: the example of heart failure. AB - Increasing amounts of medical knowledge, clinical data, and patient expectations have created a fertile environment for developing and using clinical practice guidelines. Electronic medical records have provided an opportunity to invoke guidelines during the everyday practice of clinical medicine to improve health care quality and control costs. In this paper, efforts to incorporate complex guidelines [those for heart failure from the Agency for Health Care Policy and Research (AHCPR)] into a network of physicians' interactive microcomputer workstations are reported. The task proved difficult because the guidelines often lack explicit definitions (e.g., for symptom severity and adverse events) that are necessary to navigate the AHCPR algorithm. They also focus more on errors of omission (not doing the right thing) than on errors of commission (doing the wrong thing) and do not account for comorbid conditions, concurrent drug therapy, or the timing of most interventions and follow-up. As they stand, the heart failure guidelines give good general guidance to individual practitioners, but cannot be used to assess quality or care without extensive "translation" into the local environment. Specific recommendations are made so that future guidelines will prove useful to a wide range of prospective users. PMID- 7496882 TI - Modeling the temporal complexities of symptoms. AB - OBJECTIVE: To analyze the temporal aspects of symptoms, including their temporal uncertainty, in order to develop a high-level conceptual data model representation of this domain. DESIGN: A basic tenet of existing temporal models is that events occur not only relative to a particular date or time, but also relative to the time of some other event. The time an event occurs, particularly when the event is a symptom being recalled by a patient or collected by a busy provider, is frequently incomplete or uncertain, and this uncertainty must also be represented in a temporal data model. The object-oriented modeling technique used in this study is becoming popular among U.S. medical informatics standards developers. RESULTS: A conceptual data model for the temporal aspects of symptom data, including temporal uncertainty, has been developed. The object-oriented modeling approach used enables the temporal objects and attributes defined in this model to be inherited by other medical objects, such as problems. CONCLUSIONS: The temporal comparators presented here have previously been defined, and may serve as the basis for standardizing the terms used to describe how one event temporally relates to another. In an attempt to achieve domain completeness, this study concentrated more on developing a model that is highly expressive than on developing one that is easily queried. This trade-off in representation versus "queryability" will require further analysis and may require modifications to the underlying model. PMID- 7496880 TI - The Columbia Registry of Information and Utilization Management Trials. AB - OBJECTIVE: T systematically locate, register, and abstract information used in comparing effects of various information services (computerized and noncomputerized) and utilization management interventions on the process and outcome of patient care. DESIGN: Manual and electronic database searches located reports that met three main criteria: 1) randomized controlled trial; 2) information or utilization management intervention in the study group with no similar intervention in the control group; and 3) effect of the intervention on the process and/or outcome of patient care had been measured. Published reports were registered in the Columbia Registry. RESULTS: Nearly 600 reports were collected from 24 countries and 189 different publications. Frequently tested interventions included patients or physician education, telephone follow-up, patient or physician reminders, and home care services. Frequently reported effect variables included hospitalization rate, length of stay, immunization rate, and mortality rate. Standardized formal tools were developed for the separation and abstraction of practical information and methodologic details from the collected trial reports. CONCLUSIONS: The registry provides a new source of information for meta-analyses, traditional reviews, and executive summaries of quality improvement of health services. The streamlined knowledge engineering process of quality evaluation and abstraction of critical information can generate helpful information for practitioners and researchers simultaneously. PMID- 7496883 TI - Moving ahead on webbed feet. PMID- 7496884 TI - Leveraging a fellowship in medical informatics: focus on software. PMID- 7496886 TI - Computer-algebra calculations and measurements on AB spin systems for double-spin echo sequences. AB - The time evolution of the density operator of an AB spin system during a double spin-echo pulse sequence is evaluated analytically by a computer-algebra system. The computer-algebra system allows one to generate the extensive formulas describing the density operator and yields an expression for the integral of the spectral signals. The simulation of spectra for arbitrary sequence timings can be easily performed by this new tool without risking errors that might occur in conventional calculations. The computer-algebra method can be extended straightforward to other pulse angles and types of sequences. The double-spin echo pulse sequence is used in the point-resolved spectroscopy (PRESS) method which is often applied for volume selective examinations in vivo. For verification of the results generated by the computer-algebra system, 1H spectra from a half-liter spherical sample with an aqueous solution that was 0.1 M in sodium citrate and 0.1 M in sodium acetate were recorded after 90 degrees-180 degrees-180 degrees double-spin-echo pulse sequences on a 1.5-T whole-body unit. The measured behavior of the citrate AB spin system corresponds very well with the theoretical predictions. Thus, the theory provides the basis for optimization of sequence timings for double-spin-echo measurements with high signal gain from AB systems as, for example, citrate. In addition, the theoretically predicted signal modulations could be fitted to the experimental data, providing the transverse relaxation time of the AB-coupled protons. PMID- 7496885 TI - Adapted techniques for clinical MR imaging of tendons. AB - To determine whether the echo time of magnetic resonance gradient-echo and spin echo imaging sequences may be important for the occurrence of high signal strength from tendon with pathological alterations, imaging sequences with sufficient spatial resolution and very short echo times were developed for whole body imagers with standard gradient system. The sequences were applied on the Achilles tendons of five healthy volunteers and seven patients with achillodynia. Some affected regions inside tendon, probably corresponding with tissue with subtle edema in the collagen bundles were only revealed in images recorded with very short echo times TE < 5 ms, whereas stronger affections and protons in liquids between the fiber bundles were also shown in images with longer echo times TE > 10 ms. Gradient-echo methods allow shorter echo times than spin-echo techniques for a given gradient system of the imager and given spatial resolution. So minimum echo time gradient-echo sequences should be used for sensitive imaging of tendon alterations, because no considerable signal dephasing due to susceptibility effects were found in tendon. PMID- 7496888 TI - Analysis of micellar and vesicular lecithin and cholesterol in model bile using 1H- and 31P-NMR. AB - The distribution of phospholipid and cholesterol between the vesicular and micellar phases in bile plays an important role in the formation of cholesterol gallstones. Conventional analytical procedures to determine the distribution are potentially unreliable because they disturb the distribution of these compounds between the two phases. In this work, we circumvent this problem by using NMR. 31P-NMR is used to quantify directly the micellar and the vesicular amount of lecithin. The previously described 1H-NMR method to determine directly the micellar lecithin (Groen et al., J Lipid Res 31: 1315-1321) has been optimized by the implementation of a spectral quantification method. The agreement between the 31P and 1H methods was excellent. In our hands, the method of Ellul et al. (FEBS Lett 300: 30-32) did not allow quantification of micellar cholesterol, although our fitting procedure offered the possibility of quantifying overlapping spectral peaks by the use of prior knowledge about all the parameters of the compounds visible in the spectrum. The micellar cholesterol concentration was so low compared to the overlapping lecithin peaks that no reliable quantification was possible. The same problem was encountered when using other characteristic cholesterol resonances for quantification. Our data suggest that cholesterol present in the vesicular phases is too immobile to give rise to high-resolution peaks and the amount of cholesterol present in the micellar phase is too low to allow quantitation by NMR. We conclude that 1H-NMR can be used to determine micellar lecithin, and 31P-NMR to determine micellar as well as vesicular lecithin in model bile. PMID- 7496889 TI - High-resolution magnetic resonance imaging of the endolymphatic duct and sac. AB - An anatomical study was carried out to determine the extent to which magnetic resonance imaging (MRI) could delineate inner ear structures. Anatomical preparations of human petrous temporal bone were examined and compared with the results of MRI in 20 healthy subjects to see whether the structures of the inner ear could be visualized. Imaging of the subjects was carried out in a 1.0-T MRI scanner (Siemens Magnetom Impact). Two strongly T2*-weighted sequences were used: a 3D-PSIF sequence and a 3D-CISS sequence. The 3D data sets were postprocessed using a Maximum Intensity Projection (MIP) program. Our investigations show that it is possible to obtain accurate visualization of structures with a diameter of under 1 mm. In all 20 subjects it was possible to identify both the endolymphatic duct and the endolymphatic sac. PMID- 7496890 TI - Magnetization transfer by simple sequences of rectangular pulses. AB - On-resonant radio frequency (RF) sequences composed of a train of short rectangular pulses of the same kind were optimized in order to obtain selective saturation of protons with short transverse relaxation times for magnetization transfer purposes. It is demonstrated that the sequences regarded allow a good adaptation to different requirements for magnetization transfer examinations on whole-body imagers. The sequences presented here provide relatively strong saturation of protons with very short transverse relaxation times T2 approximately less than 50 microseconds, whereas signals from protons with long T2 to be recorded are hardly influenced in a broad frequency range. The sequences are especially advantageous for applications in pulse files with limited numbers of support points. PMID- 7496887 TI - Evaluation of ultrafast phase-contrast imaging in the thoracic aorta. AB - PURPOSE: Two ultrafast phase-contrast (PC) data acquisition strategies, multishot echo-planar imaging (EPI)-PC and segmented k-space fast gradient-echo PC (FASTCARD-PC) were evaluated with regard to their measurement accuracy. MATERIALS AND METHOD: Flow measurements of the ascending and descending aorta were acquired in 10 healthy volunteers with an electrocardiogram (ECG)-triggered eight-shot EPI PC sequence (TR/TE/flip 16/7.4/45 degrees, 32-ms flow-phase interval, 2 x 2 mm in plane resolution), and FASTCARD-PC (six k-lines per band, TR/TE/flip 11/6.1/45 degrees, 132-ms flow-phase interval, 2 x 1 mm in plane resolution). These were compared to flow-volume data acquired with conventional cine-PC (TR/TE/flip 24/7/45 degrees, 48-ms flow-phase interval, 2 x 1 mm in plane resolution). Using cine-PC as a gold standard, the measurement accuracy of FASTCARD-PC and EPI-PC were determined. RESULTS: Both EPI-PC and FASTCARD-PC significantly reduced data acquisition times compared to cine PC. EPI-PC flow measurements correlated well with aortic cine-PC flow-volume determinations (r = 0.98). Reflecting poorer temporal resolution, FASTCARD-PC measurements were less accurate (p < 0.05), evidenced by poor correlation with cine-PC data (r = 0.62). CONCLUSION: Ultrafast PC measurements are possible. In contrast to the segmented k-space PC technique, which is limited in temporal resolution, multishot EPI-PC provides high measurement accuracy in pulsatile vessles while keeping the image acquisition interval short enough for a comfortable breath-hold. PMID- 7496891 TI - A gating and triggering system dedicated to nuclear magnetic resonance studies of isolated perfused heart. AB - This work reports a low-cost and versatile electronic device designed to trigger NMR acquisitions from the cardiac cycle of an isolated perfused heart, or to perform electrical stimulation of the heart. The triggering is synchronised with the pressure curve of the perfused heart. The cardiac pacing is achieved from pulses of the NMR system, or by an internal pulse generator, in order to be operated separately from the NMR instrument. PMID- 7496892 TI - T2 relaxation time study of the pig renal allografts. AB - A previous study showed the possibility of the dependence of the T2 maps measured by Siemens Magnetom MR imager 1.5 T on the viability of the cadaveric kidney. For the T2 relaxometry study the pig model was used. Ten pig renal allografts were examined during 68 h after removal. Significant differences were found in the change of the initial Tini2 values in the histogram within the first 20 h [delta 1 = Tini2 (2) - Tini2 (20)] or 8 h [delta 2 = Tini2 (2) - Tini2 (8)] after removal between the groups of the damaged kidneys (delta 1 = 20.2 +/- 6.8 ms, delta 2 = 19.4 +/- 3.0) and the kidneys without apparent damage (delta 1 = 9.4 +/ 8.5 ms, delta 2 = 5.4 +/- 5.8). PMID- 7496894 TI - Models to study the relation between oxygen consumption and oxygen delivery during an acute reduction in blood flow: comparison of balloon filling in the inferior vena cava, tamponade, and hemorrhage. AB - Different animal models have been used to study the relationship between oxygen consumption (VO2) and oxygen delivery (DO2) and each of them has its own specificity. The present study compared the models of balloon inflation in the inferior vena cava (BALL), tamponade (TAMP), and hemorrhage (HEM) to acutely reduce blood flow in pentobarbital-anesthetized dogs. The order of the procedures was randomized first, but HEM was irreversible so that HEM was applied as the third procedure (n = 10) or as a unique procedure (n = 4). Critical DO2 (DO2 crit) was determined as the intercept of two best fitted lines of regression of VO2 against DO2, but similar results were obtained when it was calculated from blood lactate or from arterio-venous differences in pH (AVpH) or PCO2 (VAPCO2). At DO2 crit, VO2 was 6.6 +/- .7 mL/min.kg in BALL, 6.2 +/- .6 mL/min.kg in TAMP and 6.0 +/- .7 mL/min.kg in HEM (p < .05 vs. BALL). DO2 crit was significantly greater with BALL (14.3 +/- 2.3 mL/min.kg) than with TAMP (10.2 +/- 1.7 mL/min.kg, p < .01 vs. BALL) or HEM (9.1 +/- 1.4 mL/min.kg, p < .01 vs. BALL). Also, critical oxygen extraction (EO2 crit) was significantly lower with BALL (46.5 +/- 4.8%) than with TAMP (62.1 +/- 11.5%, p < .01 vs. BALL) or HEM (66.8 +/ 11.4%, p < .01 vs. BALL, p < .05 vs. TAMP). The earlier onset of tissue hypoxia with BALL was probably related to the blood flow redistribution induced by this model.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7496893 TI - In vitro evidence of neutrophil-mediated lung injury after intestinal reperfusion. AB - This study examines the hypothesis that neutrophils isolated from animals sustaining intestinal reperfusion (IIR) induce pulmonary microvascular dysfunction. Lungs were isolated from normal Sprague-Dawley rats and perfused with a physiologic buffer in vitro. Neutrophils (2 x 10(6)) isolated from animals sustaining IIR (n = 5) or sham operation (SHAM; n = 6) were infused into the isolated lung model. A third group of lungs underwent in vitro perfusion without exposure to neutrophils (n = 5). Lung injury was assessed by measuring wet to dry weight ratios and pulmonary artery pressure (PAP). Pulmonary ultrastructure was assessed by electron microscopy. The wet:dry ratio of lungs from animals sustaining IIR was greater than that of lungs exposed to SHAM neutrophils (p = .03) or perfusate alone (p = .02). The PAP of lungs exposed to IIR neutrophils was nearly 10 times greater than that of lungs exposed to SHAM neutrophils (p = .003) or buffer alone (p = .006). Ultrastructural examination of lungs exposed to IIR neutrophils demonstrated interstitial edema with occasional focal disruptions in the alveolar capillary endothelial cell membrane whereas lungs exposed to SHAM neutrophils were normal. These experiments provide important in vitro correlation of prior in vivo studies suggesting that neutrophils are important pathogenic mediators of IIR-induced lung injury. PMID- 7496895 TI - Hyperosmotic glucose infusion during hemorrhage does not reduce bacterial translocation in 24 hour-starved rats. AB - Food deprivation 24 h before stress increases bacterial translocation in hemorrhage. Presently it tested whether hyperosmolality, induced by exogenous glucose infusion to improve plasma refill, prevents or reduces bacterial translocation after experimental hemorrhage in 24 h food-deprived rats. Rats were given an i.v. infusion of either 2 mL of 30% glucose (G) or the same volume of .9% NaCl (C) while simultaneously being submitted to a standardized 60 min hemorrhage period, of moderate or more severe hemorrhage. Blood was not reinfused. Despite development of marked hyperglycemia (p < .001, G vs. C) resulting in significantly greater reductions in packed cell volume (p < .001, G vs. C), bacterial translocation was detected similarly in both groups regardless of whether moderate (10/12-G, 9/12-C) or severe (15/19-G, 15/18-C) hemorrhage was inflicted. It was concluded that hyperglycemic hyperosmolality did not prevent bacterial translocation in these models of hemorrhagic stress in 24 h-starved rats. PMID- 7496896 TI - A comparison of survival at different degrees of hemorrhagic shock in germ-free and germ-bearing rats. AB - We have previously reported superior survival after one level of hemorrhagic shock in germ-free (GF) rats compared with germ-bearing (GB) rats. The objective of this study was to determine the effect of the GF state on survival at different degrees of hemorrhagic shock. GF and GB rats were bled to a mean arterial blood pressure of 30 mmHg. Shock was terminated after 10, 20, 40, or 80% of the maximum shed blood volume was reabsorbed spontaneously. Both shock time and time to decompensation were significantly longer in GF rats (p < .05). Comparative survival was greater for GF rats at most levels of shock (p < .01). This superiority in survival was greatest at moderate shock levels and decreased at severe shock levels. There may be several reasons for the increased tolerance of GF animals to hemorrhagic shock such as metabolic or immunologic variations. It is hard to avoid the fact, however, that the most notable difference between the GF and GB rat is the presence or absence of bacteria. PMID- 7496897 TI - Physiological responses, organ distribution, and circulation kinetics in anesthetized rats after hypovolemic exchange transfusion with technetium-99m labeled liposome-encapsulated hemoglobin. AB - Physiological responses and circulation properties of liposome-encapsulated hemoglobin (LEH) labeled with technetium-99m (99mTc) were measured in rats after a 10% (170 mg/kg hemoglobin; 430 mg/kg phospholipid) or a 50% (450 mg/kg hemoglobin, 2.3 g/kg phospholipid) hypovolemic exchange transfusion (n = 5 per exchange group). Mean arterial pressure returned to baseline values (105 +/- 8 mmHg) by 90 min post-infusion for both groups. By 20 h, mean arterial pressure remained at baseline values for the 10% group, but dropped to 30 +/- 14 mmHg for the 50% group. For both groups, bradycardia was seen after the exchange period, but heart rate recovered by 30 min for the 10% group and by 90 min for the 50% group. The 99mTc-LEH remained in circulation longer for the 50% group (18.2 h half-life) than for the 10% group (2.4 h half-life). Removal of 99mTc-LEH from the bloodstream was via the liver and spleen. At 20 h, 99mTc-LEH accumulation in these organs was greater for the 10% group (liver, 36.2 +/- 1.7%; spleen, 37.5 +/ 2.5%) than for the 50% group (liver, 17.0 +/- 1.4%; spleen, 17.1 +/- 1.4%). The data show that there is less clearance of 99mTc-LEH from the bloodstream by the reticuloendothelial system after a 50% hypovolemic exchange transfusion, thus supporting the possible use of LEH as an oxygen-carrying resuscitative fluid in situations of severe blood loss. PMID- 7496898 TI - Preincubation of endotoxin with monoclonal anti-lipid A (E5), but not in vivo treatment, inhibits circulatory dysfunction. AB - Monoclonal antibodies (mAb) directed against the toxic lipid A portion of lipopolysaccharide (LPS) have been shown to bind lipid A in vitro, but clinical trials of such mAbs have yielded mixed results. In 53 rats instrumented for macrocirculatory and cremaster muscle microcirculatory measurements, we examined whether E5, a murine-derived anti-lipid A mAb, could inhibit LPS-induced circulatory dysfunction when incubated with LPS in vitro or given separately in vivo prior to LPS administration. Compared with Control rats (Group I), rats infused with 10 mg/kg Escherichia coli LPS (Group II) displayed marked decreases in arterial pressure and cardiac output and marked decreases in erythrocyte velocity in second, third, and fourth order skeletal muscle arterioles. Infusion of 2 mg/kg E5 90 min prior to LPS infusion (Group III) did not improve cardiovascular performance. In contrast, incubation of LPS with either 2 mg/kg (Group IV) or 10 mg/kg (Group V) E5 prior to infusion significantly attenuated LPS-induced changes in both macrocirculatory and microcirculatory function. Further investigation of the disparity between the in vitro and in vivo neutralizing capacity of anti-lipid A mAbs may aid interpretation of the variable clinical results achieved with these preparations. PMID- 7496899 TI - Effects of endotoxin on the guinea pig heart response to ischemia reperfusion injury. AB - Ischemia causes significant damage to the heart as manifested by decreases in ventricular performance. Several different methods have been shown to protect the heart from ischemic injury--one is operative over a short period (an hour) and the other over longer periods (a day). The latter form of protection has been demonstrated in rats after induction of Gram-negative sepsis or administration of endotoxin or cytokines. In the present study we determined whether guinea pigs would also show induction of cardiac protection subsequent to a dose of endotoxin. Male guinea pigs were injected with 1 mg of endotoxin and studied the following day. Hearts were perfused at a constant perfusion pressure and studied in an isovolumic mode. Left ventricular developed pressure was significantly lower in the endotoxin-treated group than in the control group. After 35 min of total ischemia and 25 min of reperfusion, recovery of left ventricular developed pressure was complete in the endotoxin group but significantly decreased in the control group such that after ischemia and reperfusion, there was no significant difference in left ventricular performance between the two groups. Coronary flow was significantly greater in the endotoxin group than in the control group both prior to and after ischemia. Hearts from endotoxin-treated guinea pigs resumed spontaneous contractile activity sooner and released less lactate upon reperfusion than did the control group. Thus prior treatment of guinea pigs with endotoxin resulted in depression of the isolated heart but also resulted in protection of the isolated heart from further damage due to ischemia/reperfusion injury. PMID- 7496900 TI - Nitric oxide synthase inhibition is detrimental to cardiac function and promotes bronchospasm in anaphylaxis in rabbits. AB - We studied the effects of a nitric oxide synthase (NOS) inhibitor, N omega-nitro L-arginine-methyl ester (L-NAME), on cardiac depression and bronchospasm provoked by systemic anaphylaxis in vivo in rabbits. Animals pretreated with L-NAME showed lower survival rates than control animals pretreated with normal saline. The survival rate in L-NAME-pretreated animals was increased by the administration of L-arginine after initiation of anaphylaxis. Cardiac output fell significantly in animals pretreated with L-NAME compared with controls, although venous return was increased. In animals pretreated with L-NAME, pulmonary resistance was significantly increased, and administration of arginine attenuated the bronchospasm. In conclusion, these results, along with the low survival rates in the L-NAME-treated animals, suggest that NO production may be beneficial to cardiac depression and bronchospasm in anaphylaxis in vivo. PMID- 7496902 TI - Amino acid metabolism and the vascular endothelium: regulation and disease implications. AB - Amino acid metabolism by the vascular endothelium is a complex process that often begins with the carrier-mediated uptake of circulating amino acids into the endothelial cytoplasm. Amino acids are essential for maintaining intact endothelial functions, which include cell proliferation, regulation of blood flow and vascular tone, coagulation and fibrinolysis, and metabolism of a variety of macromolecules. The disturbances in endothelial amino acid transport and metabolism that occur during infection and inflammation are due, in part, to changes in substrate availability and to the local and/or systemic elaboration of specific mediators. An improved understanding of endothelial amino acid metabolism will not only provide new knowledge regarding disease mechanisms and regulation, but may also lead to new treatment strategies that may include the clinical use of specific nutritional formulas. PMID- 7496901 TI - Trauma-hemorrhage causes prolonged depression in cellular immunity. AB - A number of clinical studies have shown that multiple and severe trauma causes immunosuppression and increases the susceptibility to sepsis. However, because there is a close temporal relationship between trauma and hemorrhage in humans, it is difficult to dissociate the effects of tissue trauma versus hemorrhage on immunity in the clinical setting. Studies in mice have shown that simple hemorrhage per se as well as laparotomy alone produces a marked depression in cellular immunity and no difference was seen in the extent of depression at 2 h if these two insults were combined. Nonetheless, it remains unknown whether the combined model of trauma-hemorrhage produces a more protracted depression in immune function. To study this, 5 days after either sham operation, laparotomy (i.e. trauma), hemorrhage alone (35 mmHg for 1 h, followed by resuscitation), or the combination of laparotomy and hemorrhage, mice (C3H/HeN) were sacrificed, after which splenocyte and peritoneal macrophage cultures were established. The proliferative capacity of the splenocytes, as well as their ability to release IL 2 and IL-3, was markedly decreased in the trauma-hemorrhage animals but was normal in the other groups. Furthermore, the release of IL-6 by peritoneal macrophages from animals that underwent trauma-hemorrhage was also significantly depressed. These results support the concept that traumatic injury in the form of a midline laparotomy combined with hemorrhage produces a more protracted impairment in cell-mediated immunity than laparotomy or hemorrhage alone. PMID- 7496903 TI - Blockade of tumor necrosis factor reduces lipopolysaccharide lethality, but not the lethality of cecal ligation and puncture. AB - Inhibition of tumor necrosis factor (TNF) bioactivity has afforded protection in several animal models of sepsis. We examined whether inhibition of TNF could improve survival after lethal lipopolysaccharide (LPS) or cecal ligation and puncture (CLP) in CD-1 or BALB/c mice. Neutralizing rabbit anti-TNF antisera were evaluated in CD-1 mice by injecting the antisera 3 h before intravenous (i.v.) LPS (600 micrograms). Implantable radiotransmitters were used for continuous monitoring of temperature. No decrease in mortality was observed, and the anti TNF failed to prevent the drop in temperature. In BALB/c mice injected with antisera before LPS (200 micrograms) mortality was reduced (dead/total: control sera, 14/14; anti-TNF, 4/12; p = .007 control sera vs. anti-TNF). CD-1 mice were pretreated with anti-TNF or control sera; CLP was performed followed by administration of antibiotics. Anti-TNF did not decrease pulmonary neutrophil sequestration, improve survival, or prevent the decrease in temperature observed as sepsis developed. CLP was performed in the BALB/c mice using antibiotics plus anti-TNF antisera, but no protection was observed. Our results demonstrate that anti-TNF treatment prevents LPS mortality only when using certain strains of mice and inhibition of TNF fails to reduce mortality in a more clinically relevant model of sepsis. PMID- 7496905 TI - Simultaneous pancreas-kidney and sequential pancreas-after-kidney transplantation. AB - Simultaneous pancreas-kidney (SPK) or pancreas-after-kidney (PAK) transplantation has been advocated as an alternative to kidney transplant alone (KTA) for type 1 diabetics with end-stage renal disease. Advocates of combined transplant assert that the procedure reduces, prevents, or mitigates secondary complications of diabetes and improves the quality of life (QOL) of recipients. The combined procedures may be accomplished with a relatively low mortality, but the morbidity significantly exceeds that of KTA. The published data did not provide unambiguous support for the contention that SPK or PAK improved or ameliorated the secondary diabetic complications of retinopathy, neuropathy, and nephropathy, and it cannot be reasonably concluded that such benefit is likely to result. The majority of studies of QOL subsequent to combined transplant had significant methodologic deficiencies which made generalizations problematic. Notwithstanding, improvements in objective measures, such as return to employment or school, reduction in medical care requirements, days spent in hospital, social or physical activity, etc, have not been demonstrated for combined transplant; improvements in subjective measures were inconsistently reported. The United Network for Organ Sharing (UNOS) registry indicated that SPK represents 83 percent, and PAK about 8 percent of all pancreas transplants in the United States. Pancreas graft survival data are limited; UNOS reported 3-year survival rates of approximately 65 percent following SPK, and 35 percent after PAK. Renal graft survival following SPK appears comparable to that reported for most cadaver KTA. However, selection of SPK in lieu of KTA with a living-related donor or HLA matched cadaver kidney may result in significant reduction in expected renal graft survival, in the range of 40-70 percent to as much as 350 percent. A cost effectiveness analysis (CEA) model compared SPK with KTA and continued insulin therapy. The model employed a wide range of reported charges/payments, and postulated that SPK would provide significant improvements in quality of life. Sensitivity analyses indicated that SPK was equal in cost effectiveness to KTA only in patients who incurred very high annual costs for the treatment of hyper- or hypoglycemia. The literature does not indicate that such patients comprise the majority of SPK recipients. Additional evidence is necessary to unequivocally demonstrate the risks, costs, and ultimate benefits of combined transplant. Such information should include detailed and unambiguous patient selection criteria, prospective comparative studies of the effects of SPK/PAK upon secondary complications and quality of life, and accurate cost data for the transplant procedures and required followup care. PMID- 7496904 TI - Noninvasive evaluation of microcirculatory hemodynamic changes during hemorrhage followed by saline or blood transfusion. AB - The purpose of the study was to examine the ability of a system combining laser Doppler flowmetry (LDF), photoplethysmograph (PPG), and transcutaneous oxygen tension (tc-PO2) to follow changes in the microcirculation during hemorrhage and following blood or saline return, and to test the hypothesis that such changes precede and might predict changes in the systemic blood pressure. Measurements were performed on the skin of anesthetized rabbits (n = 10) during mild (0-8%), moderate (9-24%), and severe (25-30% of blood volume) hemorrhage, and following complete volume restitution by blood or saline. We found the following: 1) hemorrhage caused typical changes in the LDF, PPG, and tc-PO2 signals that could be formulated by mathematical models, 2) these signals identified blood as being more efficient than saline for volume restitution following hemorrhage, and 3) microcirculatory changes precede and might predict systemic hemodynamic events. PMID- 7496906 TI - Autologous peripheral stem-cell transplantation. AB - Autologous peripheral stem-cell transplantation (APSCT) has been extensively applied to support cancer patients who have undergone high-dose chemotherapy (HDCT) and suffer from the effects of otherwise prolonged or irreversible myelosuppression. The APSCT process involves harvesting of autologous progenitor cells from a patient's circulating blood (via leukapheresis), cryopreservation of the cells, and subsequent intravenous infusion for bone marrow hematopoietic reconstitution (HR). Although pluripotent stem cells, capable of multilineage differentiation, cannot be distinguished by morphologic criteria, they can be characterized as being CD34+ cells capable of indefinite self-renewal in situ and long-term self-renewal in cell cultures. Bone marrow and peripheral blood are common sources of autologous progenitor cells. Current techniques to identify and separate CD34+ cells for use in APSCT have resulted in fewer tumor cells being infused than if unseparated peripheral stem cells (PSC) were transplanted, with no differences noted in HR. Chemotherapy- and cytokine-induced mobilization results in increases in progenitor cells, necessitating fewer phereses to harvest sufficient numbers of progenitor cells for engraftment. This assessment addresses the safety, efficacy, and cost-effectiveness of the use of PSC for HR and improving patient outcome, as well as the indications and criteria for patient selection for the use of APSCT. Available information from study panels, research centers, institutions, and government agencies is reviewed; randomized clinical tests (or lack thereof) are discussed; and comparisons are made between APSCT and autologous bone marrow transplantation (ABMT), an accepted therapy in treatment of some malignancies (e.g., leukemia and lymphoma). The author concludes that existing evidence indicates that PSC can provide satisfactory HR, and the rate of HR via PSC does not seem consistently different from that of ABMT. The clinical importance of HR continues to be secondary to the primary issue of the patient benefits of HDCT in terms of antitumor response, palliation, or survival. PMID- 7496907 TI - Influence of breathing-zone ammonia on human exposures to acid aerosol pollution. PMID- 7496908 TI - Adhesion molecules in host defense. PMID- 7496909 TI - Transforming growth factor beta in Alzheimer's disease. AB - Alzheimer's disease (AD) has been hypothesized to be an inflammatory condition. We hypothesized that anti-inflammatory cytokines, such as transforming growth factor beta (TGF-beta), counteract the inflammatory process. In the present study, we found that TGF-beta levels were elevated in both cerebrospinal fluid and serum samples obtained from AD patients < 6 h after death. Serum TGF-beta levels were also markedly elevated before death. These results suggest that elevated TGF-beta levels in AD may represent a protective host response to immunologically mediated neuronal injury. PMID- 7496910 TI - Evidence for anti-Plasmodium falciparum antibodies that cross-react with human T lymphotropic virus type I proteins in a population in Irian Jaya, Indonesia. AB - This study was performed to demonstrate the presence of anti-Plasmodium falciparum antibodies in a population living in Irian Jaya, Indonesia that cross react with human T-lymphotropic virus type I (HTLV-I) proteins. Serum samples from 63 volunteers living in Oksibil, a secluded highland valley in Irian Jaya, were tested for anti-P. falciparum antibodies by an immunofluorescence assay and for anti-HTLV-I antibodies by an enzyme immunoassay (EIA). All samples were positive for anti-P. falciparum antibodies at titers of > or = 1:256. Twenty-four samples were reactive by EIA for HTLV-I, and of these, 23 were tested by western blotting (immunoblotting). Five of the 23 samples were classified as western blot positive and 18 were classified as western blot indeterminate. In competitive blocking assays with malaria proteins, western blot immunoreactivity to all HTLV I Gag proteins was either reduced or eliminated. Significant reductions in the HTLV-I EIA optical density values of the Oksibil sera occurred when the sera were competitively blocked with the malaria antigens. The optical density values of HTLV-I-positive control sera showed no significant change. Competitive blocking with HTLV-I antigens produced reductions in the optical density values of both the Oksibil sera and the HTLV-I-positive control sera. These data suggest that in this population, anti-P. falciparum antibodies are cross-reactive with HTLV-I proteins in the western blot and EIA tests. PMID- 7496911 TI - Elevated Candida antigen titers are associated with neutrophil dysfunction after injury. AB - This study was undertaken to determine if impaired neutrophil (polymorphonuclear leukocytes [PMNL]) function is associated with an elevated Candida antigen titer after injury. PMNL from eight severely injured adults with Candida antigen titers of > or = 1:4 (titer positive) were evaluated for the ability to inhibit growth of Candida albicans in vitro by using a [3H]glucose incorporation assay. PMNL from eight severely injured adults with titers of < 1:4 (titer negative) and from eight healthy volunteers were studied for comparison. PMNL from the titer positive patients had suppressed ability to inhibit C. albicans growth compared with PMNL from titer-negative patients and healthy volunteers. In vitro, PMNL function against C. albicans could be augmented significantly by cytokines. Granulocyte macrophage-colony-stimulating factor was most potent at augmenting function, followed by interleukin-8 and gamma interferon. Injured patients with elevated candida antigen titers have impaired PMNL function against C. albicans, and this function can be restored by cytokines. PMID- 7496912 TI - Persistent generalized lymphadenopathy and non-Hodgkin's lymphoma in AIDS: association with Rochalimaea henselae infection. AB - Cat scratch disease, which is caused by infection with Rochalimaea henselae, is often manifested as lymphadenopathy. R. henselae has also been isolated from human immunodeficiency virus (HIV)-positive patients with bacillary angiomatosis. In order to determine the frequency of R. henselae-reactive antibodies in HIV positive patients with persistent generalized lymphadenopathy (PGL) or non Hodgkin's lymphoma (NHL), we tested a total of 124 HIV-positive patients for R. henselae-reactive immunoglobulin G (IgG), IgM, and IgA antibodies by an enzyme immunoassay procedure using whole R. henselae antigen. Of the patients, 7 had PGL, 17 had NHL, and 100 were HIV stage IV (Centers for Disease Control criteria). A total of 86% of PGL patients (6 of 7) were positive for R. henselae antibodies (three were positive for IgG, IgA, and IgM, one was positive for IgG and IgA only, and two were positive for IgG only). A total of 29% of NHL patients (5 of 17) were positive for R. henselae antibodies (two were positive for IgG, IgA, and IgM and three were positive for IgG only). Only 5% of HIV Stage IV patients without adenopathy (5 of 100) were positive for R. henselae-reactive IgG, IgA, and IgM. The high prevalence of R. henselae-reactive antibodies in HIV positive PGL and NHL patients suggests that R. henselae is a potential etiologic agent or cofactor in these patients. PMID- 7496914 TI - Flow cytometric analysis of peripheral blood lymphocyte immunophenotypes in persons infected with Treponema pallidum. AB - To characterize the human immune response to syphilis, we determined the effect of infection with Treponema pallidum on the percentage of the various lymphocyte subpopulations in the peripheral blood of infected and uninfected persons. Monoclonal antibodies labeled with either fluorescein isothiocyanate or phycoerythrin were used to perform dual color analysis on a FACScan with the following markers: CD3 for total T cells, CD4 for T helper cells, CD8 for T suppressor cells, CD19 for B cells, and CD16 plus CD56 for natural killer cells. Lymphocyte immunophenotype results were analyzed by the stage of untreated syphilis and by gender. Although they were within the ranges of the normal distribution of immunophenotypes, the percentages of CD4+ cells were significantly lower (P < 0.001) and those of CD8+ cells were higher (P = 0.03) in patients with syphilis than in the uninfected population. For infected versus uninfected subjects, both women and men, the differences in the mean percentages of CD3+ and CD4+ cells were significant (P < or = 0.05). Significant differences were noted between the sexes in secondary syphilis only in the mean percentages of cells positive for CD3, CD4, CD8, and CD16 plus CD56. Gender had no effect on lymphocyte subpopulations in subjects with primary or latent syphilis. In the control population, significant differences due to gender were observed in the percentages of cells positive for CD3, CD4, and CD16 plus CD56. PMID- 7496913 TI - Differential elevation of circulating interleukin-1 beta, tumor necrosis factor alpha, and interleukin-6 in AIDS-associated cachectic states. AB - Elevation of serum interleukin-1 beta (IL-1 beta) levels, and to a lesser degree tumor necrosis factor alpha levels, was found in cachectic human immunodeficiency virus (HIV)-infected African patients without concurrent opportunistic infection or neoplasia (HIV wasting syndrome). A heterogeneous pattern of elevations of cytokine levels, including mild elevations of IL-1 beta and pronounced elevations of IL-6 levels, was found in other cachectic states. PMID- 7496915 TI - Increased immunogenicity of inactivated influenza virus vaccine containing purified surface antigen compared with whole virus in elderly women. AB - Thirty-eight elderly female subjects (aged 80 +/- 7 years, mean +/- standard deviation) were randomized to immunization with trivalent inactivated influenza virus vaccine containing either purified surface antigen (n = 18) or whole virus (n = 20) components from A/Texas/36/91 (H1N1), A/Beijing/353/89 (H3N2), and B/Panama/45/90 strains. Humoral and cellular immune responses were assessed by measuring serum hemagglutination inhibition antibodies and cytotoxic T lymphocyte (CTL) activity at 0 and 3 weeks postvaccination. Serological responses to both of the type A vaccine strains following immunization with surface antigen vaccine (SAV) were significantly more frequent and greater in magnitude than those induced by whole-virus vaccine. Antibody responses to the B/Panama component were modest and did not differ significantly between the two vaccines. Persons given SAV, but not those given whole-virus vaccine, had a small but significant increase in mean percent specific lysis of influenza A (H1N1) virus-infected autologous targets by peripheral blood mononuclear cells which were stimulated in vitro with influenza A (H1N1) virus. The H1N1-stimulated cytotoxic effectors induced by SAV were CD8+ and were not cross-reactive against H3N2-infected targets. Influenza B virus-specific CTL responses were not observed with either vaccine. These results suggest that currently available subunit influenza virus vaccines may offer an advantage over inactivated whole-virus preparations for inducing humoral and cellular immune responses in the elderly, although the CTL response may be too limited to be of physiological significance. PMID- 7496916 TI - Antibody response to polyribosyl-ribitol phosphate antigen of Haemophilus influenzae in Ecuadorian and German children. AB - Serum samples from 1,221 Ecuadorian children 0 to 5 years of age and from 236 German subjects were tested by enzyme-linked immunosorbent assay for class specific antibodies to the capsular polysaccharide of Haemophilus influenzae type b (PRP antigen). A gradual prevalence increase of and mean titer increase in immunoglobulin M (IgM) antibody was seen in Ecuadorian but not in German children older than 6 months. At the end of the first year of life, about 50% of the Ecuadorian children showed IgM and IgG antibody to PRP. Seroepidemiological analysis revealed that living at a low altitude and lower calorie intake (a proxy measure of breast-feeding) were factors associated with earlier acquisition of PRP antibody. Children from low-altitude areas of Ecuador also experienced significantly more episodes of significant respiratory infections. The acquisition of PRP-reactive antibodies in Ecuadorian children might thus reflect exposure to encapsulated H. influenzae type b in lower respiratory tract infections. PMID- 7496918 TI - Laboratory diagnosis of congenital syphilis by immunoglobulin M (IgM) and IgA immunoblotting. AB - We screened cord blood or serum samples from 101 infants at risk for congenital syphilis and serum samples from their mothers for immunoglobulin G (IgG), IgM, and IgA antibodies to Treponema pallidum by western blotting (immunoblotting). Clinical evaluation showed that six infants had signs and/or symptoms consistent with congenital syphilis. The sera from five of these infants were IgM blot positive, and four were IgA blot positive. Four asymptomatic infants had serologic evidence of congenital syphilis. The sera from three of these infants were IgM blot positive, and two were IgA blot positive. However, the IgM reactivity of the serum from one asymptomatic infant, which was also IgA positive, was abolished by protein G treatment. An IgM capture enzyme-linked immunosorbent assay corroborated the presence of IgM antibodies in six of seven IgM blot-reactive sera. Overall, for detection of symptomatic congenital syphilis, a sensitivity of 83% for IgM blotting and 67% for IgA blotting was obtained. The significance of positive IgM or IgA Western blots for asymptomatic infants requires further study to confirm infection in these infants. PMID- 7496917 TI - Flow cytometric immunodetection of human immunodeficiency virus type 1 proviral DNA by heminested PCR and digoxigenin-labeled probes. AB - PCR is the most sensitive and direct method for detecting blood-borne viruses, as well as an efficient means for producing vector-free probes. However, the application of PCR, especially in the laboratory diagnosis of human immunodeficiency virus (HIV) infection, is impeded by the current use of radiolabeled oligonucleotide probes. Therefore, we have developed a nonisotopic PCR immunoreactive bead (PCR-IRB) assay to detect HIV type 1 proviral DNA from peripheral blood mononuclear cells (PBMC). We used a biotinylated primer in a set of three oligonucleotides selected from the HIV long terminal repeat region for heminested PCR amplification. An internal probe was synthesized by PCR with incorporation of digoxigenin-labeled dUTP. After solution hybridization of the probe with PCR-amplified products (amplicons), the hybridized DNA was captured with streptavidin-coated magnetic beads. For the detection of hybrids, flow cytometric analyses were carried out by two procedures: (i) direct detection with fluorescein isothiocyanate (FITC)-labeled antidigoxigenin immunoglobulin G (IgG) antibody and (ii) indirect detection with antidigoxigenin sheep IgG antibody followed by FITC-labeled anti-sheep IgG antibody. Both procedures in the PCR-IRB assay detected two to three copies of HIV proviral DNA sequences, a sensitivity that is comparable with that of the conventional radioactive detection of amplicons following probe hybridization and electrophoresis. To compare the PCR IRB assay with the conventional method, we tested 53 pedigreed PBMC specimens from blood donors and newborns; the results obtained were identical. This nonisotopic PCR-IRB assay can also be automated for potential application in laboratory diagnosis of HIV infection, blood bank screening, and therapeutic monitoring of viremia and perinatal transmission. PMID- 7496919 TI - Autoimmune hemolytic anemia in chronic mucocutaneous candidiasis. AB - Chronic mucocutaneous candidiasis is an immunodeficiency disease characterized by T-cell dysregulation and chronic superficial candidal infections. We report on three patients with chronic mucocutaneous candidiasis who developed autoantibodies to erythrocytes. Our first patient, a 19-year-old female, developed autoimmune hemolytic anemia (AIHA) that required multiple courses of treatment, including corticosteroids, intravenous immunoglobulin, and danazol. During the last exacerbation of AIHA, intensive treatment with corticosteroids and intravenous immunoglobulin failed and yet the patient responded to plasmapheresis. Our second patient, a 21-year-old male, developed AIHA which responded to oral corticosteroid therapy. Our third patient, a 6-year-old female without evidence of hemolysis, was found to have erythrocyte autoantibodies on routine screening. These three patients had positive direct antiglobulin tests, and the first patient had both immunoglobulin G (IgG) and IgM erythrocyte autoantibodies, while the remaining two patients had only IgG autoantibody. This is the first report of the association of AIHA with chronic mucocutaneous candidiasis. We suggest that all patients with chronic mucocutaneous candidiasis be screened periodically for erythrocyte autoantibodies. Plasmapheresis, a safe ancillary procedure in the management of AIHA, may be life-saving in some cases. The occurrence of erythrocyte autoantibodies in mucocutaneous candidiasis may be related to immunoregulatory disorders in this disease. PMID- 7496920 TI - Detection of borreliacidal antibody by using acridine orange and flow cytometry. AB - Borreliacidal antibody has been shown to be important for the serodiagnosis of Lyme disease and determination of immune status. Our results show that borreliacidal antibody can be rapidly and accurately detected by flow cytometry. Acridine orange was added to normal and immune sera containing Borrelia burgdorferi organisms in the presence and absence of complement prior to data acquisition by flow cytometry. The flow cytometric parameters of side scatter and detection of acridine orange fluorescence were used to determine events per minute (number of labeled spirochetes), percent shift in fluorescence (number of dead spirochetes), and mean channel fluorescence (intensity of fluorescence labeled spirochetes) of acridine orange-labeled spirochetes. Borreliacidal antibody was detected as early as 4 h, with optimal detection 16 to 24 h after incubation of B. burgdorferi organisms with immune serum and complement. Our results also showed that complement was necessary for detection of borreliacidal antibody. Flow cytometry with acridine orange-labeled spirochetes provides a rapid means for detection of borreliacidal antibody. PMID- 7496922 TI - Production, characterization, and application of monoclonal antibodies to Vibrio cholerae O139 synonym Bengal. AB - Mouse monoclonal antibodies (MAbs) were derived against acetone-treated whole cells of the newly recognized Vibrio cholerae O139 serogroup which is causing epidemics of cholera-like disease in India and Bangladesh. Four MAbs specifically recognized the lipopolysaccharide antigens of V. cholerae O139. MAbs ICL9 and ICL13 were of the immunoglobulin M (IgM) isotype, ICL11 was of the IgG3 isotype, and ICL12 was of the Ig2b isotype. A fifth MAb, ICL10, of the IgG2b isotype cross reacted with V. cholerae O91. All five MAbs recognized V. cholerae O139 in an enzyme-linked immunosorbent assay, slide agglutination test, motility inhibition test, and indirect immunofluorescence test. During a 1-month evaluation of these MAbs in our clinical laboratory, all 86 cases diagnosed as V. cholerae O139 by a rabbit polyclonal antiserum were also detected by these MAbs, establishing their utility as highly sensitive and specific diagnostic reagents. With these MAbs, it should now be possible to screen for the V. cholerae O139 serogroup in epidemic and endemic diarrhea cases and in environmental and food samples. PMID- 7496921 TI - Immunologic cross-reactivity between structural proteins of human T-cell lymphotropic virus type I and the blood stage of Plasmodium falciparum. AB - To determine the serologic cross-reactivity between human T-cell lymphotropic virus type I (HTLV-I) and parasite antigens, we measured antibody responses against HTLV-I, Plasmodium falciparum, Plasmodium vivax, and Brugia malayi in serum specimens obtained from regions where malaria (n = 482) and filariasis (n = 101) are endemic. Analysis of immune reactivity to HTLV-I antigens showed that specimens from regions where malaria is endemic had significantly higher rates of enzyme immunoassay (EIA) reactivity (76 of 482 [15.8%] than those from regions where filariasis is endemic (0 of 101 [0%]). Western blot (immunoblot) analysis of the HTLV-I EIA-reactive specimens demonstrated predominant Gag reactivity (HTLV-Iind). Only two specimens each from Indonesia and Brazil and four specimens from Papua New Guinea had Env reactivity by radioimmunoprecipitation analysis. Furthermore, a positive correlation between HTLV-EIA and titers of antibody to the blood stage of P. falciparum (rs = 0.24, P < 0.005) was discerned; no correlation was observed between antibodies to the blood stage or the circumsporozoite protein of P. vivax and the circumsporozoite protein of P. falciparum. In addition, P. falciparum-infected erythrocyte lysate specifically abrogated binding of Gag-specific antibodies in HTLV-Iind specimens from regions where malaria is endemic without affecting binding in HTLV-I-seropositive specimens, suggesting that the immunologic cross-reactivity between HTLV Gag proteins and malaria parasites is restricted to the blood-stage antigens of plasmodia in specimens from regions where malaria is endemic. However, HTLV seroindeterminate specimens from the United States did not demonstrate serologic cross-reactivity, suggesting that antigenic mimicry of HTLV proteins extends to other nonplasmodial antigens as well. PMID- 7496923 TI - Appropriate coating methods and other conditions for enzyme-linked immunosorbent assay of smooth, rough, and neutral lipopolysaccharides of Pseudomonas aeruginosa. AB - Smooth, rough, and neutral forms of lipopolysaccharide (LPS) from Pseudomonas aeruginosa were used to assess the appropriate conditions for effective enzyme linked immunosorbent assay (ELISA) of LPS. Each of these forms of well-defined LPS was tested for the efficiency of antigen coating by various methods as well as to identify an appropriate type of microtiter plate to use. For smooth LPS, the standard carbonate-bicarbonate buffer method was as efficient as the other sensitivity-enhancing plate-coating methods compared. The rough LPS, which has an overall hydrophobic characteristic, was shown to adhere effectively, regardless of the coating method used, to only one type of microtiter plate, CovaLink. This type of plate has secondary amine groups attached on its polystyrene surface by carbon chain spacers, which likely favors hydrophobic interactions between the rough LPS and the well surfaces. Dehydration methods were effective for coating microtiter plates with the neutral LPS examined, which is composed predominantly of a D-rhamnan. For the two dehydration procedures, LPS suspended in water or the organic solvent chloroform-ethanol was added directly to the wells, and the solvent was allowed to dehydrate or evaporate overnight. Precoating of plates with either polymyxin or poly-L-lysine did not give any major improvement in coating with the various forms of LPS. The possibility of using proteinase K- and sodium dodecyl sulfate-treated LPS preparations for ELISAs was also investigated. Smooth LPS prepared by this method was as effective in ELISA as LPS prepared by the hot water-phenol method, while the rough and neutral LPSs prepared this way were not satisfactory for ELISA. PMID- 7496924 TI - Monoclonal antibodies against a 97-kilodalton antigen from Aspergillus flavus. AB - We prepared a panel of five monoclonal antibodies (MAbs) directed against Aspergillus flavus that all reacted against one 97-kDa antigen by western blot (immunoblot). Flow cytometry demonstrated that these antibodies bound (in increasing degrees) to all morphologic stages of A. flavus growth: conidia, swollen conidia, and hyphae. Cross-reactivity among species was examined by enzyme-linked immunosorbent assay of fungal culture filtrates. Four MAbs reacted with 10 of 11 A. flavus isolates, and the fifth one reacted with 9 of them. One MAb also reacted with A. fumigatus, two reacted with A. niger, A. wentii, and A. nidulans, and all five reacted with A. ochraceus. None reacted with A. terreus, A. glaucus, A. versicolor, or a Penicillium species. Each MAb bound to A. flavus hyphae in formalin-fixed paraffin sections of a muscle biopsy from a confirmed human case of invasive aspergillosis. In summary, these MAbs identified a 97-kDa antigen found on A. flavus that is both surface bound and an exoantigen. Either the same or a cross-reacting antigen is present in A. fumigatus and other Aspergillus species. PMID- 7496925 TI - Detection of beta-2 transferrin in otorrhea and rhinorrhea in a routine clinical laboratory setting. AB - A simple, straightforward, and rapid method for the detection of beta-2 transferrin in otorrhea and rhinorrhea that can be used in a routine clinical laboratory is described. The beta-2 transferrin was detected by agarose gel electrophoresis of the fluid on Beckman Paragon equipment, followed by pressure transfer to a nitrocellulose membrane and then incubation with enzyme-labeled antitransferrin antibody and substrate. The procedure was fast (3.5 h) and sensitive (detected as little as 1 microgram/ml) and required only 3 microliters of fluid. Beta-2 transferrin was detected in cerebrospinal fluid diluted up to eightfold. No special training or expertise was needed, and all equipment and procedures used are commonly available in a routine clinical laboratory. PMID- 7496926 TI - A confocal microscopy study of anticytoskeletal antibody activity in patients with connective tissue disease. AB - The significance of the presence of antibodies to cytoskeleton proteins in patients with connective tissue diseases is not clear, as there is a high level of these antibodies in healthy controls. In an attempt to improve the visualization of the immunofluorescence binding pattern of autoantibodies to cytoskeletal structures in cultured fibroblasts, we have used confocal microscopy. Of the 256 serum samples tested, 155 (61%) WERE reactive with cytoplasmic structures. These reactive samples could be divided into seven patterns of binding, as determined by double-blind examination of single-section confocal images. While confirming the results of previous immunofluorescence studies which have shown that autoantibodies that bind to filamentous structures in the cytoplasm of cultured cells are common in patients with connective tissue diseases, we were able to identify three patterns of cytoskeletal binding which may be useful as an adjunct to other tests for the diagnosis of some connective tissue diseases, in particular systemic sclerosis (scleroderma) and rheumatoid arthritis/Sjogren's syndrome. None of the seven patterns was exclusive to a particular disease. We conclude that confocal microscopy may be of limited use as an adjunct to other serological assays in the diagnosis of some forms of connective tissue disease. PMID- 7496927 TI - Enzyme-linked immunosorbent assay using a recombinant baculovirus-expressed Bacillus anthracis protective antigen (PA): measurement of human anti-PA antibodies. AB - We developed an antigen capture enzyme-linked immunosorbent assay (ELISA) which does not require purified protective antigen (PA) for detection of human antibodies to Bacillus anthracis PA. Lysates of Spodoptera frugiperda (Sf-9) cells infected with recombinant baculovirus containing the PA gene were used as the source of PA to develop the ELISA. Recombinant PA from crude Sf-9 cell lysates or PA purified from B. anthracis Sterne strain was captured by an anti-PA monoclonal antibody coated onto microtiter plates. We demonstrated that human serum antibody titers to PA were identical in the ELISA whether we used crude Sf 9 cell lysates containing recombinant baculovirus-expressed PA or purified Sterne PA. Finally, false-positive results observed in a direct ELISA were eliminated with this antigen capture ELISA. Thus, the antigen capture ELISA with crude preparations of baculovirus-expressed PA is reliable, safe, and inexpensive for determining anti-PA antibody levels in human sera. PMID- 7496929 TI - Neutralizing antibody immune response in children with primary and secondary rotavirus infections. AB - We have characterized the neutralizing antibody immune response to six human rotavirus serotypes (G1 to G4, G8, and G9) in Brazilian children with primary and secondary rotavirus infections and correlated the response with the G serotype of the infecting rotavirus strain. Twenty-five children were studied: 17 had a single rotavirus infection, 4 were reinfected once, and 4 experienced three infections. Two of the reinfections were by non-group A rotaviruses. Among the 25 primary infections, we observed homotypic as well as heterotypic responses; the serotype G1 viruses, which accounted for 13 of these infections, induced mostly a homotypic response, while infections by serotype G2 and G4 viruses induced, in addition to the homotypic, a heterotypic response directed primarily to serotype G1. Two of the primary infections induced heterotypic antibodies to 69M, a serotype G8 virus that by RNA electrophoresis analysis was found not to circulate in the population during the time of the study. The specificity of the neutralizing antibody immune response induced by a virus of a given serotype was the same in primary as well as secondary infections. These results indicate that the heterotypic immune response induced in a primary rotavirus infection is an intrinsic property of the virus strain, and although there seem to be general patterns of serotype-specific seroconversion, these may vary from serotype to serotype and from strain to strain within a serotype. PMID- 7496928 TI - Monoclonal antibodies that distinguish between encephalitogenic bovine herpesvirus type 1.3 and respiratory bovine herpesvirus type 1.1. AB - Seven mouse hybridoma cell lines producing monoclonal antibodies (MAb) against an encephalitogenic strain of bovine herpesvirus type 1.3 (BHV-1.3) were established. The clones producing MAb were selected to be specific for BHV-1.3 by enzyme-linked immunosorbent assay. Only L1B neutralized virus without complement. With the addition of complement, five of the MAb neutralized BHV-1.3 but not the respiratory strain BHV-1.1. The anti-BHV-1.3-specific MAb Q10B, L6G, and L1B precipitated glycoproteins from BHV-1.3 that were analogous to the gI, GIII, and gIV glycoproteins of BHV-1.1, respectively. The other four MAb precipitated unknown proteins. None of the anti-BHV-1.3 MAb precipitated BHV-1.1 glycoproteins. The majority of the anti-BHV-1.3 MAb did not react with BHV-1.1 by immunoblotting, but O7E (unknown protein pattern by radioimmunoprecipitation) was reactive with five proteins (M(r)s of 33,000, 43,000, 70,000, 141,000, and 190,000) of BHV-1.3 and with a different pattern of proteins of BHV-1.1 (M(r)s of 30,000, 38,000, 83,000 and 144,000). Two of the MAb, L6G and O7E, conjugated with peroxidase were found to be useful for detecting BHV-1.3 antigen by immunochemistry in Formalin-fixed brain tissue from experimentally infected calves. PMID- 7496930 TI - Immune response to bovine viral diarrhea virus induced by anti-idiotypic antibodies. AB - We have previously prepared rabbit anti-idiotypic antibodies (anti-Ids) against the neutralizing monoclonal antibodies (MAbs) specific for the gp53 of bovine viral diarrhea virus (BVDV). The anti-Ids, purified by sequential immunoaffinity chromatography, inhibited the immunizing MAb from binding to the original antigens and blocked BVDV infection of cell cultures. This study evaluated immune responses in mice to the purified anti-Id reagents. BVDV-specific neutralizing antibodies were induced by the anti-Ids. The antisera (Ab3) induced by the anti Ids immunoprecipitated gp53 from BVDV-infected Madin-Darby bovine kidney (MDBK) cell lysates. However, lymphocyte-proliferative responses were specific only for the respective immunizing antigens. These results suggest that the anti-Ids may bear an internal image of the gp53 to stimulate production of antibody but not to stimulate a virus-specific cellular immune response in mice. PMID- 7496931 TI - Murine splenic hematopoietic subpopulations: the enlarged undifferentiated subset in New Zealand black mice is multipotent stem cells. AB - We recently reported that a significant population of the murine splenic non-T, non-B "null" cell compartment consists of non-lineage-specific, undifferentiated cells which are in the G0 and G1 phases of the cell cycle and that their numbers are particularly high in the spleens of New Zealand Black mice. A highly enriched population of these non-lineage-specific cells obtained by successive elimination of differentiated cells was further purified to homogeneity by fluorescence activated cell sorting. The morphologic, phenotypic, and histochemical characteristics of this purified population suggest that these cells may be primitive hematopoietic stem cells. The germ line configuration of the genomic DNA establishes that these are uncommitted stem cells. In vivo, these cells form day 12 colonies in the spleen and liver of lethally irradiated recipients and confer radioprotection. These cells also differentiate into T- and B-cell lineages and reconstitute the immunodeficiency in mice with severe combined immunodeficiency. In response to a combination of a very few early-acting lymphokines and/or stromal cell-conditioned medium in vitro, these cells differentiate into both myeloid and lymphoid cell types. More of these cells are obtained from the enlarged spleens of New Zealand Black mice than from those of BALB/c mice. The presence of a comparatively higher number of stem cells in the spleen than in the marrow or fetal liver provides an alternative, and possibly superior, source of uncommitted stem cells for a variety of experimental investigations or therapeutic manipulations. PMID- 7496933 TI - Vaccine immune response and side effects with the use of acetaminophen with influenza vaccine. AB - The purpose of this study was to determine whether acetaminophen impairs the immune response to influenza vaccine. Influenza vaccine is an under-utilized preventive measure, partly because of the unfounded perception that fever and myalgias frequently follow vaccination. While acetaminophen may decrease these infrequent side effects, it may also alter the immune response to vaccination. We compare the effect of acetaminophen with placebo on the humoral immune response to the 1991-1992 commercially available influenza vaccine. We studied 60 healthy, elderly subjects from a geriatric clinic and 20 infirm, elderly subjects from a nursing home. The subjects were randomly assigned to receive placebo or acetaminophen (1,000 mg every 6 h) for 2 days. Acetaminophen did not depress or enhance the immune development of serum hemagglutination inhibition antibody to the three vaccine antigens. The systemic side effects of fever and myalgia were uncommon in both groups. The healthy elderly subjects mounted a significantly better immune response to the influenza virus A/Taiwan/1/86 (H1N1) vaccine strain than did the infirm elderly subjects (geometric mean titer, 115 versus 51; P = 0.003). The functional activity score obtained by using the chronic healthy evaluation component of the Acute Physiology and Chronic Health Evaluation system could be used to distinguish the healthy from the infirm elderly (scores of 1.27 versus 3.75, P < 0.001). Acetaminophen neither depressed nor enhanced the serum antibody response to the vaccine in the healthy and infirm elderly subjects studied. PMID- 7496932 TI - Role of human natural killer cells in health and disease. AB - Natural killer (NK) cells, the CD3- CD56+ CD16+ subset of peripheral blood lymphocytes, have long been known to be involved in non-major histocompatibility complex-restricted natural immunity to virally infected and malignant target cells. The association of abnormalities in NK cell numbers or functions with a broad spectrum of human diseases has been more clearly defined in recent years as a result of the improved knowledge of NK cell physiology and advances in monitoring of NK cell functions in health and disease. The ability to reliably measure changes in NK activity and/or numbers during the course of disease or response to treatment has focused attention on the role of the NK cell in disease pathogenesis. The improved understanding of NK cell deficiency in disease has opened a way for therapies specifically designed to improve NK cell function. The therapeutic use of biologic response modifiers capable of augmenting NK cell activity in vivo and of adoptive transfer of highly enriched, activated autologous NK cells in diseases such as cancer and AIDS is being evaluated. The importance of NK cells in health and the consequences of NK cell deficiency or excess are likely to be more extensively monitored in the future. PMID- 7496935 TI - Differentiation of borreliacidal activity caused by immune serum or antimicrobial agents by flow cytometry. AB - We demonstrated that borreliacidal activity caused by immune serum and complement can easily be differentiated by flow cytometry from killing activity caused by antimicrobial agents that are commonly used for the treatment of Lyme disease. Assay suspensions containing normal or immune serum were incubated with Borrelia burgdorferi in the presence or absence of ceftriaxone, doxycycline, penicillin, and phosphomycin for 2, 8, 16, and 24 h. Samples containing killing activity were identified by using flow cytometry and acridine orange. In 30 min, the effects of immune serum and complement were easily distinguished from the killing of spirochetes by antimicrobial agents by adding fluorescein isothiocyanate conjugated goat anti-hamster immunoglobulin. This simple procedure greatly enhanced the usefulness of the borreliacidal assay by eliminating a major source of false-positive reactions. PMID- 7496934 TI - Immunological analysis of the components of the antigen complex A60 of Mycobacterium bovis BCG. AB - The antigen complex of A60 of Mycobacterium bovis BCG was analyzed by different immunological techniques to assess its relevance to tuberculosis and the involvement of its components in the immune reactions elicited in humans by tuberculous infection. A60 is composed of about 30 components, of which 8 were identified by available monoclonal antibodies (lipoarabinomannan, a glycolipid, and proteins of 65, 40, 38, 35, 19, and 14 kDa). The majority (87.5%) of anti mycobacterial antibodies in sera from tuberculosis patients was directed against A60. Western blot (immunoblot) analysis indicated that the majority of the highly antigenic proteins present in mycobacterial homogenates were components of the A60 complex. A small percentage (7.8%) of A60 epitopes proved to be species specific. Thus, A60 proteins of 66, 41, 38, 37, 35, 34, 32, and 22 kDa were found to contain B-cell epitopes specific for M. bovis and not shared by Mycobacterium leprae oR Mycobacterium avium. PMID- 7496936 TI - Immune response in the lungs following oral immunization with bacterial lysates of respiratory pathogens. AB - We have investigated the local immune response of the BALB/c mouse respiratory tract after oral immunization with a bacterial lysate of seven common respiratory pathogens. After two immunization on five consecutive days, we examined the immunoglobulin (immunoglobulin G [IgG], IgM, and IgA) secretion rates of cells isolated from the lungs and compared them with those of spleen cells of orally immunized and nonimmunized animals by using a new test system based on time resolved fluorescence. The procedure followed the principle of the classical ELISPOT test with nitrocellulose-bottomed microtiter plates, but europium (Eu3+) linked streptavidin rather than enzyme-conjugated streptavidin was used, with the advantage of quantifying secreted immunoglobulins instead of detecting single antibody-secreting cells. Lymphocytes isolated from the lungs of treated animals revealed significant increases in total and antigen-specific IgA synthesis compared with the rates of the controls, whereas IgG and IgM production rates showed no remarkable differences. In addition, the sera of treated mice revealed higher antigen-specific IgA titers but not increased IgM and IgG levels. We conclude that priming the gut-associated lymphoid tissue with bacterial antigens of pneumotropic microorganisms can elicit an enhanced IgA response in a distant mucosal effector site, such as the respiratory tract, according to the concept of a common mucosa-associated immune system. PMID- 7496937 TI - Clinical significance of serum 2,5-oligoadenylate synthetase and soluble interleukin-2 receptor in hemophiliacs positive and negative for human immunodeficiency virus type 1. AB - We measured serum 2,5-oligoadenylate synthetase (2,5-AS) levels and soluble interleukin-2 receptor (sIL-2R) levels in human immune deficiency virus type 1 (HIV-1)-positive and HIV-1-negative hemophiliacs in order to clarify the clinical significance of these parameters in hemophiliacs. Serum 2,5-AS levels were measured by a radioimmunosorbent assay, and sIL-2R levels were measured by an enzyme-linked immunosorbent assay. The mean serum 2,5-AS levels were higher in AIDS-related-complex and AIDS patients, asymptomatic carriers, and HIV-1-negative hemophiliacs than in hepatitis C virus-positive patients and healthy controls. Serial determinations showed that the 2,5-AS levels tended to increase in HIV-1 positive patients, especially those with AIDS-related complex or AIDS, although it showed a substantial decrease in the terminal stage. The serum sIL-2R levels were higher in HIV-1-positive patients, HIV-1-negative patients, and hepatitis C virus-positive patients than in controls. Serial studies showed little change in the HIV-1-positive and HIV-1-negative groups, although sIL-2R levels showed a tendency to decrease with zidovudine treatment. On the basis of the present results, we may well conclude that 2,5-AS and sIL-2R are not specific markers for hemophiliacs with HIV-1 infection. However, serial measurement of these markers can still be useful for assessing the progression of AIDS and the prognosis for patients with AIDS, as well as for monitoring the response to zidovudine. PMID- 7496938 TI - Presence of hepatitis C virus (HCV) genomic RNA and viral replicative intermediates in bone marrow and peripheral blood mononuclear cells from HCV infected patients. AB - The cellular tropism of hepatitis C virus (HCV) was studied in vivo in samples from patients with persistent HCV infection. Plasma, liver, peripheral blood mononuclear cell (PBMC), and bone marrow cell (BMC) samples from 15 subjects positive for anti-HCV antibodies were tested for the presence of HCV RNA sequences by reverse transcription PCR. Virus-specific RNA sequences were found to be present in liver samples from all subjects (100%), in plasma samples from 13 of 15 patients (86.7%), in PBMC samples from 3 patients (20%), and in BMC samples from 9 (60%) of the 15 anti-HCV-positive patients enrolled in this study. The presence of the molecular intermediate of HCV replication (the negative stranded HCV RNA) was evident in the two of the three PBMC and in five of the nine BMC HCV RNA-positive samples. Finally, we studied the nucleotide sequence of a large portion (-270 to -59) of the 5'untranslated region of HCV amplified from plasma samples of 12 of the 15 patients with and without HCV in BMCs; the degree of heterogeneity compared with the prototype HCV sequence was similar in both groups. The data principally indicate that HCV infection of PBMCs and BMCs is frequent in persistently infected patients, as shown by the occurrence of positive- and negative-stranded HCV RNA, thus suggesting the possibility of extrahepatic replication of HCV. PMID- 7496939 TI - Novel in vitro method for identification of individuals at risk for beryllium hypersensitivity. AB - Beryllium-specific lymphocytes were generated by in vitro immunization of peripheral blood mononuclear cells (PBMC) from healthy unexposed individuals. Measurement of blastogenic responses of PBMC by [3H]thymidine uptake demonstrated that sensitization of PBMC with beryllium salts followed by stimulation with unrelated salts resulted in a negative response, whereas sensitization and restimulation of PBMC with beryllium salts produced a positive response. Flow cytometric and cell depletion analyses showed that all of the responding cells were CD4+ T cells. The in vitro immunization system was used to screen 52 human subjects for susceptibility to beryllium sensitization in vitro. The results show that of the 52 healthy unexposed subjects tested, only 1 (2%) was highly responsive, 4 subjects (8%) were moderately responsive, 20 subjects (39%) were low-level responders, and 27 subjects (52%) were nonresponders. The results showing 2% high-level responsiveness to beryllium sensitization in vitro correlate with the 1 to 5% prevalence of chronic beryllium disease in individuals sensitized to beryllium dust in vivo and thus support the thesis that the in vitro immunization system may permit the identification of individuals at risk for beryllium hypersensitivity. PMID- 7496941 TI - Detection of antibodies to trans-activator protein (p40taxI) of human T-cell lymphotropic virus type I by a synthetic peptide-based assay. AB - Antibodies to human T-cell lymphotropic virus type I (HTLV-I) trans-activator protein (p40taxI) were determined in serum specimens from individuals infected with HTLV-I (n = 138) and HTLV-II (n = 19). Western blot (immunoblot) analysis using recombinant tax demonstrated the presence of anti-tax antibodies in 96% of patients (25 of 26) with HTLV-I-associated myelopathy, 43% of those (20 of 46) with adult T-cell leukemia, and 61% of asymptomatic HTLV-I blood donors (40 of 66); only one of the HTLV-II specimens reacted with the recombinant tax protein. Synthetic peptides (Tax8(106-125), Tax22(316-335), Tax-23(331-350), and Tax 24(336-353) representing the immunodominant epitopes of p40taxI detected anti-tax antibodies in 66 (48%), 50 (36%), 66 (48%), and 64 (46%) of 138 HTLV-I-positive specimens, respectively. An enzyme immunoassay using an equimolar ratio of these four peptides allowed sensitive detection of anti-tax antibodies in 96% of patients (25 of 26) with HTLV-1-associated myelopathy, 52% of adult T-cell leukemia patients (24 of 46), and 62% of asymptomatic HTLV-1-infected donors (41 of 66). The synthetic peptide-based cocktail assay was HTLV-I specific, since none of the HTLV-II-infected specimens reacted with these peptides. Interestingly, the corresponding regions from the HTLV-II tax protein, Tax8II(106 125), and Tax-22II(312-331) did not react with either HTLV-II or HTLV-I specimens. Thus, a synthetic peptide-based assay composed of immunodominant epitopes located towards the amino terminus and the C terminus of p40taxI provides a reliable and sensitive assay for the detection of anti-tax antibodies in seroepidemiologic studies. PMID- 7496940 TI - Effect of vitamin A supplementation on immunoglobulin G subclass responses to tetanus toxoid in children. AB - Previously, we demonstrated that administering vitamin A supplements to children resulted in a significant increase in the immunoglobulin G (IgG) response generated against a vaccine dose of tetanus toxoid (TT) (R. D. Semba et al., J. Nutr. 122:101-107, 1991). However, from these analyses we could not determine whether there was an increase in levels of IgG of the subclass presumed to be important for protection against challenge by the toxin or whether there was simply a general increase in the levels of all the IgG subclasses expressing anti TT activity. The goal of this study was to determine the profile of the anti-TT IgG subclasses in children receiving vitamin A supplementation or a placebo in order to assess the potential utility of the enhanced anti-TT response. In a randomized, double-masked, placebo-controlled clinical trial, the levels of the different anti-TT IgG subclasses were measured in 139 Indonesian preschool children (3 to 6 years of age) 2 weeks before and 3 weeks after immunization. Baseline anti-TT levels and immunization histories were used to separate those children who were responding to TT for the first time from those who responded in a secondary fashion because of previous exposure to TT. Children who were given vitamin A prior to immunization had significant increases in IgG1 levels regardless of whether they were undergoing primary or memory reactions. In the group of individuals who underwent a secondary response to TT, vitamin A supplementation was also associated with a modest but significant change in the levels of anti-TT IgG3. There were only minor changes in the levels of anti-TT IgG2 and IgG4. Since IgG1 is the subclass associated with a protective response to TT immunization, these results suggest that vitamin A supplementation may be a safe and effective intervention to enhance the relevant humoral response to TT and other vaccine antigens. PMID- 7496942 TI - Serum antibody responses of divers to waterborne pathogens. AB - To assess the significance of exposure of divers to waterborne pathogens, specific immunoglobulin G serum antibody responses to Pseudomonas and Aeromonas isolates recovered from dive sites from the respiratory tracts of nine experienced divers and seven diving trainees working in the Chesapeake Bay area over a 6- to 18-month period were measured. A significant increase in the frequency of isolation of these organisms from respiratory surfaces both groups of divers after each dive was noted, with the divers' ears being the predominant recovery site (48%; P < 10(-8), chi-square). The acute serum responses of the majority of experienced divers (83%) showed evidence of preexisting antibody to these potential pathogens, whereas the acute serum response of only 32% of naive divers showed such evidence (P < 10(-8), chi-square). Six months into their training, the rate of seroresponse of the trainees to organisms recovered after their first dives increased to 61% (P = 0.003, chi-square), suggesting that repeated exposure in necessary for generation of a specific systemic immunologic response. The rate of acquisition of a new seroresponse to recovered organisms was approximately 12% per dive for both groups of divers, suggesting that there is continuous exposure to, and infection with, new strains present in the water during dives. These data suggest that, in cases in which systemic antibody is important for protection, there are various levels of susceptibility to waterborne potential pathogens in both experienced and inexperienced divers. PMID- 7496943 TI - Comparison of three commercially available assays for detection of varicella zoster virus antibody. AB - Three commercially available diagnostic assays for the detection of antibodies to varicella-zoster virus were evaluated to determine which would be the most suitable for our clinical laboratory. Three different methods were examined: an enzyme-linked immunosorbent assay (ELISA), latex agglutination (LA), and an indirect fluorescent-antibody technique. For the 141 serum specimens tested, the ELISA had an agreement of 90.1% and LA had an agreement of 92.2% with the indirect immunofluorescent-antibody technique. The ELISA had a lower sensitivity (85.6%) than LA (100.0%), but suffered from a low specificity (78.4%) compared with the ELISA (98.0%). PMID- 7496944 TI - Q fever serology: cutoff determination for microimmunofluorescence. AB - Q fever, a worldwide zoonosis caused by Coxiella burnetii, lacks clinical specificity and may present as acute or chronic disease. Because of this polymorphism, serological confirmation is necessary to assess the diagnosis. Although microimmunofluorescence is our reference technique, the cutoff titers that are currently used to make a diagnosis of active or chronic Q fever were determined years ago with limited series of patients and sera. We determined the titers of immunoglobulin G (IgG), IgM, and IgA against both phases (I and II) of Coxiella burnetii. Rheumatoid factor was removed before testing IgM and IgA. We report here the various cutoff titers and the kinetics of antibody development from 2,218 first serum samples of patients, among whom 208 suffered from acute Q fever and 53 had chronic Q fever. In active Q fever, we have defined a low cutoff (phase II IgG titer < or = 100) below which the diagnosis cannot be made and would need further confirmation and confirmed a high cutoff (phase II IgG titer > or = 200 and phase II IgM titer > or = 50) over which the diagnosis can be made. For chronic Q fever diagnosis, phase I IgA titers are not contributive despite previous works claiming their usefulness; a phase I IgG titer of > or = 800 is highly predictive (98%) and sensitive (100%). We have also studied the possibility of rejecting or evoking the diagnosis of chronic Q fever by phase II IgG and IgA titers. This method is useful when phase I testing is not available, but the sensitivity remains low (57%). PMID- 7496945 TI - Flow cytometric measurement of immunoglobulin E to natural latex proteins. AB - Immediate hypersensitivity to natural latex (NL) occurs in sensitized individuals after repeated exposure to products or devices containing NL components. Since allergic reactions to NL proteins are quite frequent and may be quite serious, diagnostic assays are needed to identify individuals at risk. A number of latex proteins have been considered the major antigens, but they have been incompletely characterized. There is no standard material available for skin testing. In vitro diagnostic tests, such as the radioallergosorbent test (RAST), are time consuming and their sensitivity and specificity remain to be proven. We have developed a rapid microsphere-based, fluorescence-activated flow cytometry assay for the measurement of NL protein-specific human immunoglobulin E and have compared it with both the enzyme-linked immunosorbent assay and radioallergosorbent test methods. By using the total purified NL protein fraction isolated from raw ammoniated NL sap as the antigen, the flow cytometry assay was both sensitive and specific for the detection of NL protein-specific human immunoglobulin E in the sera of sensitized pediatric patients. PMID- 7496946 TI - Decline in delayed-type hypersensitivity response in obese women following weight reduction. AB - The effect of weight loss on immuno function was evaluated in 14 middle-aged obese women. Cutaneous delayed-type hypersensitivity to five recall antigens was measured before a weight loss of 21 kg and again after weight had been stabilized. Weight loss was associated with a decrease in both number and magnitude of delayed-type hypersensitivity responses. The number of positive skin tests decreased from 3.1 +/- 0.2 to 2.4 +/- 0.3 per subject, and the mean cutaneous induration size decreased by 38%. The decrease in induration size was positively correlated with the decrease in body mass index (r = 0.63), weight (r = 0.59), and fat mass (r = 0.52). These preliminary findings suggest that large weight losses may result in decreased expression of immune function. PMID- 7496947 TI - Immunochemical identification of Brucella abortus lipopolysaccharide epitopes. AB - Sera from Brucella abortus-infected and -vaccinated bovines recognized four lipopolysaccharide (LPS) determinants: two in the O-polysaccharide (A and C), one in the core oligosaccharide from rough Brucella LPS (R), and one in lipid A (LA). From 46 different hybridomas secreting monoclonal antibodies (MAbs) against various LPS moieties, 9 different specificities were identified. Two epitopes, A and C/Y, were present in the O-polysaccharide. Two epitopes were found in the core oligosaccharide (R1 and R2) of rough Brucella LPS. MAbs against R1 and R2 epitopes reacted against LPS from different rough Brucella species; however, MAbs directed to the R2 epitope also reacted against enterobacterial LPS from deep rough mutants. Three epitopes (LA1, LA2, and LA3) were located in the lipid A backbone. Different sets of MAbs recognized two epitopes in the lipid A associated outer membrane protein (LAOmp3-1 and LAOmp3-2). LPS preparations from smooth brucellae had small amounts of rough-type LPS. Although LPS from rough brucellae did not show smooth-type LPS in western blots (immunoblots), two hybridomas generated from mice immunized with rough B. abortus produced antibodies against smooth B. abortus LPS. Results are discussed in relation to the structure and function of B. abortus LPS and to previous findings on the epitopic density of the molecule. PMID- 7496948 TI - Identification and characterization of Neospora caninum tachyzoite antigens useful for diagnosis of neosporosis. AB - The purpose of the present study was to identify antigens of the protozoan Neospora caninum that could be useful for the diagnosis of neosporosis in domestic animals. As revealed by immunoblotting, immune sera from a wide range of animal species exhibited a similar recognition pattern of four major and several minor N. caninum antigens. In contrast to preinoculation sera, all tested immune sera recognized nonreduced immunodominant 17-, 29-, 30-, and 27-kDa antigens. A 46-kDa protein which showed faint recognition by preimmune sera also exhibited a strong response by immune sera. Immunolocalization of the four immunodominant N. caninum antigens was investigated by immunogold electron microscopy using monospecific polyclonal antisera. The 17-kDa antigen appears to be associated with the body part of the rhoptries, while the 29- and 30-kDa antigens were associated with the dense granules, network, and limiting membrane of the parasitophorous vacuole. Studies were also conducted to compare antibody responses to N. caninum and the related protozoan Toxoplasma gondii. Although N. caninum and T. gondii (RH strain) tachyzoites shared a few cross-reacting antigens, the immunodominant antigens of both parasites were not recognized by heterologous sera. Also, immunogold staining with rabbit anti-Neospora hyperimmune serum exhibited almost no labeling of external membranes of Neospora tachyzoites compared with the very marked labeling seen when Toxoplasma tachyzoites (RH strain) were incubated with rabbit anti-Toxoplasma hyperimmune serum. These unique antigenic differences should be useful in developing a diagnostic assay for N. caninum. PMID- 7496950 TI - Primary and secondary granule release by polymorphonuclear leukocytes exposed to peritoneal dialysis effluent. AB - Peritoneal dialysis effluent from patients with end-stage renal failure contains a low-molecular-weight solute that inhibits the killing of phagocytosed Staphylococcus epidermidis by polymorphonuclear leukocytes (PMN). This observation has been investigated by using luciginen-enhanced chemiluminescence to measure PMN NADPH oxidase activity, CD11b/CD18 expression and lactoferrin release to measure secondary granule discharge, and cellular levels of beta glucuronidase (EC 3.2.1.31) to measure changes in primary granules. Peritoneal dialysis effluent had no effect on the loss of intracellular beta-glucuronidase from normal unstimulated PMN or from PMN stimulated with S. epidermidis. It did, however, cause a concentration-dependent (0 to 70%; vol/vol) increase in expression of CD11b/CD18 and NADPH oxidase activity. CD11b/CD18 expression increased over 20 min before starting to plateau. Release of lactoferrin by the same cells demonstrated a strong positive correlation with integrin expression (P < 0.001, Spearman's rank correlation coefficient). When dialysis effluent-treated PMN were stimulated with formyl-methionylleucylphenylalanine, integrin expression, release of lactoferrin, and NADPH oxidase activity were greater than in PMN treated with formyl-methionylleucylphenylalanine alone. Under these conditions, a concentration-dependent increase in CD11b/ CD18 and lactoferrin release were observed only at a concentration between 0 and 30% (vol/vol) dialysis effluent, while a concentration-dependent increase in oxidase activity was seen at a concentration between 0 and 70% (vol/vol). The results suggest that dialysis effluent does not affect PMN primary granule release but does cause increased release of secondary granules and an increase in NADPH oxidase activity in both unstimulated and stimulated PMN. PMID- 7496951 TI - Effect of specimen storage on absolute CD4 counts. AB - We have evaluated the effect of specimen storage on absolute CD4 counts by a commercially available manual assay. This assay utilizes latex particles coated with CD4 monoclonal antibodies that are mixed with lymphocytes in whole blood. Thirty blood samples were analyzed on days 1, 2, 4, and 7 postcollection. Linear regression analysis and Pearson correlation coefficients were used to determine the relationship between the absolute CD4 count and the storage time after sample collection. There was a significant decrease in absolute CD4 counts from baseline over time, dropping 3.6% at day 2, 10.1% at day 4, and 18.8% at day 7. However, the standard error of the B coefficient was constant [SE (B) = 0.031] up to day 4, indicating that reliable estimates of the baseline CD4 counts could be made from the CD4 counts determined up to day 4 from the time of sample collection. In addition to being sample, rapid, and inexpensive, the manual assay is capable of giving a reliable absolute CD4 count after specimen storage of up to 4 days. The application of this assay in the limited facilities of developing countries' laboratories is attractive. PMID- 7496952 TI - Lack of antibody response to invasin in humans with yersiniosis. AB - The Yersinia pseudotuberculosis inv gene encodes invasin, a 103-kDa outer membrane protein allowing bacteria to penetrate mammalian cells. This protein is produced in vitro at below 30 degrees C. In this work, we studied the antibody response against invasin in humans suffering from yersiniosis and in mice orally infected with a virulent strain of Y. pseudotuberculosis. Infection with enteropathogenic Yersinia strains did not induce either a systemic or a gut antibody response to invasin. Our results suggest that the inv gene is not expressed in the gut at 37 degrees C and, therefore, that invasin is not present to the immune system when microorganisms multiply in the host tissues. PMID- 7496949 TI - Effects of mild exercise on cytokines and cerebral blood flow in chronic fatigue syndrome patients. AB - Chronic fatigue syndrome (CFS) is an idiopathic disorder characterized by fatigue that is markedly exacerbated by physical exertion. In the present study, we tested the hypothesis that mild exercise (walking 1 mph [1 mile = 1.609 km] for 30 min) would provoke serum cytokine and cerebral blood flow abnormalities of potential pathogenic importance in CFS. Interleukin-1 beta, interleukin-6, and tumor necrosis factor alpha were nondetectable in sera of CFS patients (n = 10) and healthy control subjects (n = 10) pre- and postexercise. At rest, serum transforming growth factor beta (TGF-beta) levels were elevated in the CFS group compared with the control group (287 +/- 18 versus 115 +/- 5 pg/ml, respectively; P < 0.01). Serum TGF-beta and cerebral blood flow abnormalities, detected by single-photon emission-computed tomographic scanning, were accentuated postexercise in the CFS group. Although these findings were not significantly different from those in the control group, the effect of exercise on serum TGF beta and cerebral blood flow appeared magnified in the CFS patients. Results of this study encourage future research on the interaction of physical exertion, serum cytokines, and cerebral blood flow in CFS that will adopt a more rigorous exercise program than the one used in this study. PMID- 7496953 TI - Comparative analysis of antibodies to Francisella tularensis antigens during the acute phase of tularemia and eight years later. AB - Approximately 8 years after treatment for tularemia, 14 of 22 (63.6%) individuals tested still had a positive microagglutination test for Francisella tularensis antibodies. An enzyme-linked immunosorbent assay for anti-F. tularensis outer membrane antibodies was positive for 55% (immunoglobulin A [IgA]), 95% (IgG), and 27% (IgM) of the late-phase sera, but with antibody levels significantly reduced from those in the acute-phase sera. IgG and IgA antibody levels in the late-phase sera showed significant correlation with levels in the acute-phase sera. The IgG/IgM ratio calculation discriminated between acute-phase and persistent antibodies for most sera, but Western blot (immunoblot) patterns did not. Immunoblotting indicated that the F. tularensis lipopolysaccharide is a major target for antibodies in both groups of sera. Our results substantiate the need for caution in the interpretation of positive serological test results for tularemia, which could result from disease occurring years earlier. PMID- 7496954 TI - Occurrence, in Crohn's disease, of antibodies directed against a species-specific recombinant polypeptide of Mycobacterium paratuberculosis. AB - Sera from patients with Crohn's disease and control were analyzed by an enzyme linked immunosorbent assay based on the Mycobacterium paratuberculosis-specific recombinant polypeptide a362. Anti-a362 immunoglobulin G (IgG) (P < 0.05) and IgA (P < 0.001) titers were higher in patients with Crohn's disease than in controls. A monomodal Gaussian distribution of anti-a362 IgA levels were found for controls, and a bimodal distribution was found for patients with Crohn's disease. An M. paratuberculosis etiology is suggested for the 36% of patients with Crohn's disease who had an anti-a362 IgA level higher than that of controls. PMID- 7496955 TI - Augmentation of anti-influenza virus hemagglutinin antibody production by Peyer's patch cells with Bifidobacterium breve YIT4064. AB - Bifidobacterium breve YIT4064 was tested an an adjuvant for oral influenza vaccine by the murine Peyer's patch cell culture method. The organism augmented production of anti-influenza virus hemagglutinin immunoglobulin A antibody by Peyer's patch cells in response to addition of hemagglutinin. These antibodies may be disseminated to the respiratory mucosal tissue and prevent influenza virus infection. PMID- 7496956 TI - Demonstration of human immunoglobulin G Fc-binding activity in oral bacteria. AB - Nonimmune binding of immunoglobulins via the Fc fragment may reduce opsonization and phagocytosis of bacteria and is thus considered a virulence factor. The aim of this study was to investigate a wide range of oral bacterial strains for the presence of human immunoglobulin G (IgG) Fc-binding activity. A total of 132 strains representing 40 different gram-positive and gram negative bacterial species were tested for IgG Fc-binding activity by using a fast and simple dot blot procedure with horseradish peroxidase-conjugated Fc fragments from human IgG. Neither the human nor animal biotype of Porphyromonas gingivalis possessed IgG Fc-binding activity. The strongest positive reaction of gram-negative species with the IgG Fc fragments were obtained with strains of Prevotella intermedia and Fusobacterium nucleatum. Among the gram-positive bacteria tested, Peptostreptococcus micros, Lactobacillus spp., and several species of streptococci possessed IgG Fc-binding activity. In the present investigation, the ability of several oral bacterial species to bind IgG Fc fragments was demonstrated. This factor represents a potential virulence determinant as it may help pathogenic oral bacteria escape host defense mechanisms. PMID- 7496959 TI - Cytokines and cytokine measurements in a clinical laboratory. PMID- 7496958 TI - Expression of hepatitis E virus putative structural proteins in recombinant vaccinia viruses. AB - Hepatitis E virus (HEV) is a polyadenylated, positive-stranded RNA virus which is a major cause of enterically transmitted non-A, non-B hepatitis in many developing countries. The viral genome contains three different open reading frames (ORFs): ORF1, which is believed to encode nonstructural proteins, and ORF2 and ORF3, which are believed to encode structural proteins. The full-length putative structural proteins encoded by ORF2 and ORF3 of HEV have been cloned and expressed in recombinant vaccinia virus. Proteins encoded by ORF2 and ORF3 when expressed in vaccinia virus are recognized by pooled sera obtained from individuals with acute hepatitis E. Vaccinia-expressed viral gene products of HEV will have utility in characterizing the cell-mediated immune response to HEV. PMID- 7496957 TI - Salmonella-specific monoclonal antibodies against recombinant Salmonella typhi 36 kilodalton porin. AB - Mouse monoclonal antibodies were raised against recombinant Salmonella typhi 36 kDa porin monomer. Specificities of 16 monoclonal antibodies were analyzed as reactivity patterns in dot immunobinding and Western blot (immunoblot) assays using isolated outer membrane proteins of gram-negative bacteria and cloned purified S. typhi porin monomers and trimers. Four monoclonal antibodies were specific for Salmonella spp. PMID- 7496960 TI - In vitro cytokine production by normal human peripheral blood mononuclear cells as a measure of immunocompetence or the state of activation. AB - Measurements of cytokine levels in serum may not adequately reflect the cytokine producing potential of immune cells because of the short half-lives of cytokines and the presence of various inhibitors in human sera. In vitro cytokine production by peripheral blood mononuclear cells (PBMCs) can be an important and reliable measure of immunocompetence. Also, spontaneous in vitro release of cytokines by PBMCs may serve as a measure of their activation in vivo. In the present study, normal ranges for the in vitro production by PBMCs of interleukin 1 beta (IL-1 beta), tumor necrosis factor alpha (TNF-alpha), IL-2, and gamma interferon (IFN-gamma) were established; the feasibility of using cryopreserved PBMCs for assays of in vitro cytokine production was evaluated; and spontaneous (unstimulated) versus induced production of cytokines by fresh and cryopreserved PBMCs from healthy donors was compared. Supernatants obtained from paired fresh and frozen PBMCs were quantitated for IL-1 beta, TNF-alpha, IL-2, and IFN-gamma by using enzyme-linked immunosorbent assay or a radioimmunoassay standardized against World Health Organization cytokine standards. Fresh or cryopreserved PBMCs activated with lipopolysaccharide produced comparable levels of IL-1 beta. However, the mean levels of stimulated production of TNF-alpha, IFN-gamma, and IL 2 were significantly higher in cryopreserved versus fresh PBMCs (P < or = 0.0004). Correlations between the level of production of each cytokine by fresh versus cryopreserved in vitro-stimulated PBMCs were statistically significant, although of moderate magnitude. Spontaneous cytokine release by fresh versus cryopreserved cells was not significantly different. PMID- 7496962 TI - Spontaneous lymphocyte proliferation in human T-cell lymphotropic virus type I (HTLV-I) and HTLV-II infection: T-cell subset responses and their relationships to the presence of provirus and viral antigen production. AB - Spontaneous lymphocyte proliferation (SLP) during in vitro culture of mononuclear cells (MCs) characterizes over half of asymptomatic individuals infected with human T-cell lymphotropic virus type I (HTLV-I) or HTLV-II. Both CD4 and CD8 T cell subsets within MC cultures are activated during SLP, as judged by high density CD25 (CD25bright) expression; it is unclear, however, whether both cell subsets can directly undergo SLP. In the present investigation, the SLP capacities of purified CD8 and CD4 cells were examined in subjects infected with HTLV-I (n = 19) or HTLV-II (n = 54) in relation to the SLP status of MCs from each subject. No increase in SLP was observed for CD8 or CD4 cells from SLP negative (SLP-) HTLV-infected subjects, whereas robust SLP characterized CD8 cells from all SLP-positive (SLP+) individuals, regardless of HTLV type. In contrast, SLP+ CD4 cells characterized only 23% (7 of 31) of HTLV-II+ SLP+ individuals, whereas SLP+ CD4 cells characterized 100% of HTLV-I+ SLP+ individuals. In cocultures of HTLV-II+ SLP+ CD8 cells and autologous SLP- CD4 cells, sizable proportions of both CD8 cells and CD4 cells coexpressed CD25bright, suggesting that SLP- CD4 cells were activated in the presence of SLP+ CD8 cells. PCR analysis for tax sequences detected provirus in most CD4- and CD8 cell preparations from HTLV-seropositive individuals, regardless of type and the SLP status of cell subsets. To determine whether SLP was associated with activation of viral genes, levels of HTLV-I and HTLV-II core antigen (Ag) in supernatants were measured. Viral Ag production and SLP responses were significantly correlated for both CD4 and CD8 cells in both HTLV-I and HTLV-II infections. However, inhibition of CD8- or CD4-cell SLP by cyclosporin A or anti Tac (anti-CD25) did not reduce Ag production, indicating that Ag production is not coupled to SLP. These findings show that CD4 cells from SLP+ HTLV-I+ and SLP+ HTLV-II+ individuals differ in SLP capacity, that the absence of SLP does not indicate a lack of infection, and that production of viral Ag is associated with, but not dependent on, SLP. PMID- 7496961 TI - Elevated levels of total and Puumala virus-specific immunoglobulin E in the Scandinavian type of hemorrhagic fever with renal syndrome. AB - In a previous study, it was reported that the total immunoglobulin E (IgE) level was increased in patients with hemorrhagic fever with renal syndrome (HFRS). The aim of the present study was to investigate whether specific IgE is synthesized during the course of the disease. For this purpose, an epsilon-capture enzyme linked immunosorbent assay was developed. A total of 72 patients with HFRS caused by Puumala virus were studied. Three different control groups were included: 20 blood donors, 20 patients with other viral diseases (influenza A and B virus, acute Epstein-Barr virus, and acute cytomegalovirus infections), and 5 subjects with high levels of total IgE (median, 1,070 kU/liter; range, 773 to 5,740 kU/liter). The levels of total IgE were significantly higher during the acute phase of HFRS than those of blood donors (P < 0.01) and of patients with other viral diseases (P < 0.001). All patients developed a specific IgE response (median, 55 arbitrary units; range 24 to 123 arbitrary units) in the acute phase of the disease, whereas in the different control groups no specific IgE was detectable. Both total and specific IgE levels decreased during convalescence compared with those during the acute phase of HFRS (P < 0.001 and P < 0.001, respectively). In conclusion, we have shown that both total and specific IgE levels are increased in patients with HFRS compared with levels in patients with other viral diseases. The possible pathogenetic role of the specific IgE response in HFRS is discussed. PMID- 7496963 TI - Use of fixed autologous stimulator cells to correctly present human immunodeficiency virus type 1 viral peptides to nonhuman primate lymphocytes in proliferation and cytotoxic T-lymphocyte assays. AB - Autologous, virus-transformed lymphoblastoid cell lines were established by using peripheral blood lymphocytes from rhesus monkeys that were previously immunized with recombinant human immunodeficiency virus type 1 strain IIIB glycoprotein 160. These autologous cell lines were used to present human immunodeficiency virus type 1 viral antigens in a processed and cell-associated manner to T lymphocytes. This was accomplished by either infecting the cells with recombinant vaccinia viruses or pulsing them with synthetic peptides and then subjecting them to a mild fixation step with glutaraldehyde. Fixed antigen-presenting cells were then used as stimulator cells in vitro to measure cell-mediated immune responses. Both the vaccinia virus-infected and peptide-pulsed autologous cells stimulated antigen-specific cellular proliferative responses. The magnitude of the responses correlated with the immunization histories of the animals and other measures of immunity, such as antibody titers. Autologous vaccinia virus-infected cells were also capable of inducing the in vitro maturation of CD4+ and CD8+ precursor cytotoxic T lymphocytes into antigen-specific mature cytotoxic T lymphocytes. The use of stimulator cells to present viral peptides in a cell-associated manner appeared to be a very sensitive and versatile manner in which to measure cell mediated immune responses with peripheral blood lymphocytes from nonhuman primates. It is likely that a similar approach will function with peripheral blood lymphocytes from humans. PMID- 7496964 TI - Antigenic structure of the complete nonstructural (NS) 2 and 5 proteins of hepatitis C virus (HCV): anti-HCV NS2 and NS5 antibody reactivities in relation to HCV serotype, presence of HCV RNA, and acute HCV infection. AB - Antigenic regions within the nonstructural (NS) 2 and 5 proteins of hepatitis C virus (HCV) were identified and characterized by the use of 127 overlapping synthetic peptides and a serum panel consisting of 167 human serum samples from persons with antibodies to HCV. Initially, 20 anti-HCV-positive serum samples were used to screen the peptides covering the complete NS2 and NS5 proteins. Among the 27 overlapping peptides spanning the NS2 protein of HCV, only the peptide covering residues 960 to 975 was recognized by human sera. Within the 100 peptides covering the NS5 protein, major linear antigenic regions were located at residues 2284 to 2329 within the putative NS5a and at residues 2584 to 2599 and 2944 to 2959 within the putative NS5b. Additional minor linear antigenic regions were also identified within the NS5. The sequence of the antigenic region of the NS2 protein is, unlike most parts of the NS2 protein, highly conserved among the described types of HCV, whereas the sequence of the major antigenic region of NS5 shows variability among HCV types. The recognition of a peptide corresponding to a part of the major region of NS5 was found to be dependent on HCV type. In 129 anti-HCV-positive serum samples, the prevalence of antibodies to the NS2 protein was found to be 23% among HCV RNA-positive sera and 10% among HCV RNA-negative sera. In the same samples, reactivity to the major linear antigenic regions of HCV NS5 was found in 68% of the HCV RNA-positive sera and 67% of the HCV RNA negative sera. Of 18 serum samples from five patients with acute HCV infections, and who seroconverted with respect to anti-HCV, 4 were found to be reactive to one or more of the 100 NS5 peptides and in three serum samples the NS5 reactivities were found to shorten the time for serodiagnosis of cross-reactive with a region form residues 2584 to 2599 of NS5, which has 67% homology with a six-residue sequence of NS2. In conclusion, in this study we have identified and evaluated the potential use of synthetic peptides corresponding to linear antigenic regions of the NS2 and NS5 proteins. PMID- 7496965 TI - Lipopolysaccharide-reactive immunoglobulin E is associated with lower mortality and organ failure in traumatically injured patients. AB - Antilipopolysaccharide (anti-LPS) immunoglobulin G (IgG) and IgM have been associated with protection from LPS effects in vivo. We investigated the presence of IgE and anti-LPS in 32 patients that had experienced severe traumatic injury and in 35 healthy volunteers; we also investigated whether IgE anti-LPS was associated with important clinical events. Plasma samples were collected daily from patients in the intensive care unit and on one occasion from volunteers; the samples were assayed for IgE anti-LPS. IgE anti-LPS was assayed by enzyme-linked immunosorbent assay with monoclonal anti-human IgE as the capture antibody. Detection was accomplished with biotin-labeled LPS (Escherichia coli J5 mutant) followed by streptavidin-peroxidase with 2,2'-azino(3 ethylbenzthiazoline)sulfonic acid as the substrate. The assay was demonstrated to be specific for IgE and LPS-biotin by nonreactivity of control sera with high titer anti-LPS IgG and IgM and by inhibition with unlabeled LPS. IgE anti-LPS was detected in 1 of 35 healthy controls (2.9%) and 25 of 32 traumatically injured patients (78%) (P < 0.001). The presence of IgE anti-LPS was associated with a lower incidence of death (P = 0.026) and of renal failure (P = 0.0012). There was no apparent temporal relationship between detection of IgE anti-LPS and clinical events. IgG anti-LPS was detected more frequently in patients that were positive for IgE anti-LPS (P = 0.06) but was not associated with clinical events. The inability to detect IgE anti-LPS may be related to adverse clinical events through depletion of specific IgE due to LPS exposure after trauma or through saturation of the assay by IgE with other specificities. We have reported increased total IgE concentrations in these patients. (J.T. DiPiro, R.G. Hamilton, T. R. Howdieshell, N. F. Adkinson, and A. R. Mansberger, Ann. Surg. 215:460-466, 1992). PMID- 7496966 TI - A comparative study of frozen-section immunoperoxidase and flow cytometry for immunophenotypic analysis of lymph node biopsies. AB - Immunophenotyping by flow cytometry and frozen-section immunoperoxidase was compared on 21 consecutive lymph node biopsy specimens, of which a diagnosis of lymphoma was made for 11 specimens. Samples for flow cytometry were obtained by a fine-needle aspiration technique. Concordance between frozen-section immunoperoxidase and flow cytometry for all routine markers on all specimens ranged from 76 to 100%. In general, B-cell markers showed poorer concordance than T-cell markers, with kappa and lambda light chains having the poorest concordance, at 76% each. Flow cytometry was significantly more sensitive (90 versus 30%; P < 0.006) and had a significantly higher negative predictive value (100 versus 63%; P < 0.006) than frozen-section immunoperoxidase for demonstrating light-chain restriction. There was no significant difference in the specificities (100 versus 91%) or positive predictive values (100% each) between the two methods. Both methods demonstrated characteristic immunophenotypes for intermediate cell lymphomas, small lymphocytic lymphomas, and T-cell lymphoblastic lymphomas. Frozen-section immunoperoxidase and flow cytometry appear to be significantly concordant methods for immunophenotypic analysis of lymph node biopsies. Light-chain restriction is more readily demonstrated by flow cytometry than frozen-section immunoperoxidase. We believe that ex vivo fine needle aspiration is a simple and reliable method of obtaining cell suspensions of lymph nodes for flow cytometry. PMID- 7496967 TI - T-cell responses to infected autologous monocytes in patients with cutaneous and mucocutaneous leishmaniasis. AB - Although there is strong evidence that the control and resolution of human leishmanial infections depend primarily on activation of parasite-infected macrophages mediated by lymphokines derived from T cells, less is known about the nature of the responding cell type(s) which is protective or the antigen(s) (Ag[s]) that elicits these cells to respond. Studies using preparations of whole soluble Ag ("dead Ag") show that patients respond to a wide range of leishmanial Ags. The objective of the present study was to characterize the response of T cells from patients with healing or healed cutaneous or mucosal infections to Ag expressed by or derived from actively infected autologous monocytes ("live Ag"). Unfractionated T cells proliferated and produced gamma interferon in response to both live and dead Ags. Depletion of CD4+ T cells resulted in the loss of proliferative and gamma interferon responses to both live and dead Ags. The effect of CD8 depletion, although variable and not limited to the cells stimulated by infected monocytes, was clear for some patients. Expansion of T cells specific for live Ags by using amastigote-infected cells followed by restimulation with fast-protein liquid chromatography-fractionated soluble Ags revealed that a diversity of Ags are associated with infected monocytes. There may, however, be quantitative differences in the expression of certain Ags since prestimulation with live Ag induced higher responses to restimulation in mucocutaneous leishmaniasis patients than in localized cutaneous leishmaniasis patients. Prestimulation with dead Ag induced similar secondary responses in both patient groups. PMID- 7496968 TI - Serum antibody response to the superficial and released components of Helicobacter pylori. AB - Superficial and released components were extracted from six selected Helicobacter pylori strains. The protein and antigenic profiles of these extracts were representative of the profiles found most frequently among the clinical strains and included major peptidic fractions at 19, 23.5, 57, 68, 76, 118, and 132 kDa and major antigens at 68, 57, and 23.5 kDa. Immuno-cross-reactions were seen with a hyperimmune rabbit serum to Campylobacter fetus but not with sera to Campylobacter jejuni or Salmonella spp. An antigenic preparation was obtained by pooling equivalent quantities of each extract, and the antigenic preparation was used to study the antibody responses of sera from 65 French patients and 127 Tunisian patients. By enzyme-linked immunosorbent assay, we observed that the sera from French and Tunisian patients clustered into two populations, defined as antibody positive (72 patients) and antibody negative (120 patients). The antibody-positive patients were more frequently infected with H. pylori (P < 0.01) and were more frequently affected with gastritis (P = 0.05). However, no correlation between antibody levels and clinical signs of dyspepsia was noticed. The proportions of antibody-positive patients were similar in France and Tunisia. Antibody-positive and antibody-negative sera were studied by western blot (immunoblot) analysis. The antibody-positive sera revealed an average of 7.7 antigenic bands, whereas the antibody-negative sera revealed an average of 2.4 antigenic bands (P < 0.01). The antigens between 15 and 40 kDa and greater than 66 kDa were specifically recognized by the antibody-positive sera, although in this molecular size range the antibody profiles of these sera exhibited a fairly high degree of diversity. We conclude that the superficial and released components from H. pylori contain a variety of bacterial immunogens and may be useful in antigenic preparations for the serodiagnosis of H. pylori infections. Moreover, a group of antigens in combination appears to be useful for discriminating antibody-positive and antibody-negative patients. PMID- 7496972 TI - Molecular basis for serological cross-reactivity between enteroviruses. AB - The recognition sites for human antibodies which are cross-reactive between different types of enteroviruses were determined and characterized. Serum samples obtained from 58 patients with culture-confirmed enteroviral infections were analyzed in enzyme immunoassays against two sets of overlapping synthetic peptides covering residues 31 to 96 of poliovirus 1 VP1 (Mahoney strain) and residues 31 to 148 of coxsackievirus B1 VP1 (position based on alignment with poliovirus 1 VP1, Mahoney strain). A major antigenic region eliciting cross reactive antibodies could be located to residues 37 to 51 of VP1. Furthermore, a single peptide covering residues 42 to 55 almost completely inhibited the binding of human antibodies to heat-inactivated enteroviruses, indicating that residues 42 to 55 of VP1 contain a major region eliciting cross-reactive antibodies. By using peptide analogs in which each residue within positions 42 to 55 of VP1 was sequentially substituted by Ala or Gly, we were able to determine the most essential residues for human antibody binding in 38 of the convalescent-phase patient serum samples. In a majority of the serum samples, the most essential residues for antibody binding were found to be Pro-42, Ala-43, Leu-44, Thr-45, Ala-46, Glu-48, Thr-49, and Gly-50. All of these residues are conserved, according to known enterovirus sequences, with the divergent echovirus 22 excepted. In conclusion, we could demonstrate that the essential residues for binding of cross-reactive antibodies are well conserved within the enterovirus family. These findings provide a molecular basis for the observed antibody cross reactivity within the enterovirus group. PMID- 7496969 TI - Serological response of patients suffering from primary and recrudescent typhus: comparison of complement fixation reaction, Weil-Felix test, microimmunofluorescence, and immunoblotting. AB - Microimmunofluorescence and Western immunoblotting were compared with the classical complement fixation reaction and the Weil-Felix test to study the serological responses of patients to Rickettsia prowazekii and both Proteus vulgaris OX19 and OX2 during primary and recrudescent typhus infections. The serological response to R. prowazekii was found to be similar during primary and recrudescent typhus, and all sera examined contained antibodies to the same R. prowazekii cell structures. Immunoglobulin G (IgG) and IgM were found to be the dominant anti-R. prowazekii immunoglobulins in all sera tested and were found to be directed against the 100-kDa protein and the lipopolysaccharide. IgA antibodies, when present, were mainly against the 100-kDa protein. For P. vulgaris, IgG antibodies recognized the proteins and lipopolysaccharides of both OX19 and OX2 serotypes; IgM antibodies were directed against the P. vulgaris OX2 lipopolysaccharide. In addition, donor blood sera, which were negative by microimmunofluorescence, were found to contain IgG immunoglobulins reacting with R. prowazekii protein antigens of 135, 60, and 47 kDa by western immunoblotting. PMID- 7496970 TI - Correlation between serological and mucosal inflammatory responses to Helicobacter pylori. AB - In 82 patients who underwent gastroduodenoscopy, acute and chronic gastric mucosal inflammation was scored for severity, and systemic humoral immune responses to Helicobacter pylori antigens were assessed by enzyme-linked immunosorbent assays. On the basis of culture, gastric histology, and serologic evaluation, 33 patients were classified as H. pylori infected and 36 were classified as uninfected. Thirteen patients had negative cultures and stains but were seropositive and were analyzed separately from the other two groups. Specific serum immunoglobulin G (IgG) subclass responses to H. pylori whole-cell antigens and specific IgG responses to the 54-kDa heat shock protein homolog (Hp54K) and vacuolating cytotoxin were significantly greater in infected than in uninfected patients as were specific IgA responses to whole-cell antigens and cytotoxin (P < 0.001). Among the H. pylori-infected persons, serum IgG responses to Hp54K and to the vacuolating cytotoxin were correlated with acute mucosal inflammatory scores. In contrast, serum IgA responses to whole-cell sonicate and to vacuolating cytotoxin were inversely related to chronic inflammatory scores. By multivariant regression analysis, only specific serum IgG responses to Hp54K correlated with severity of inflammation (both acute and chronic; P < 0.001); these responses may be markers of inflammation or these antibodies could play a direct role in the pathogenesis of H. pylori-induced inflammation. PMID- 7496971 TI - Characterization of substance P binding to human monocytes/macrophages. AB - Substance P (SP), a member of the tachykinin family of neuropeptides, can immunomodulate human T cells and monocytes. SP has been shown to stimulate human monocytes to produce inflammatory cytokines and superoxide ions, and it enhances tumoricidal activity in vitro. A specific SP receptor, however, has not been identified on human monocytes/macrophages. In this study, we report that 125I-SP binds to human monocytes/macrophages with high affinity and specificity (Kd = 2.7 x 10(-8) to 5.5 x 10(-8) M). Our measurements of binding affinity to this single class of receptors were possible only when experiments were performed in the presence of excess serine proteinase inhibitor (serpin) enzyme complex receptor ligand. We determined that 125I-SP bound to a specific receptor on human monocytes/macrophages and that this binding was detectable as early as 6 h and was maintained throughout 6 to 8 weeks in culture. Modulation of the diverse immunological and inflammatory effects of SP on human monocytes may be mediated through this specific SP receptor. PMID- 7496973 TI - Comparison of cytotoxic properties of neonatal and adult neutrophils and monocytes and enhancement by cytokines. AB - We studied cytotoxic capabilities of newborn polymorphonuclear leukocytes (PMNs) and monocytes and their enhancement by cytokines and antibodies. Umbilical cord PMNs were assessed for their ability to kill various target cells spontaneously, after activation with phorbol myristate acetate, in the presence of antiserum (antibody-dependent cellular cytotoxicity), and in the presence of dually specific antibody (heteroantibody-mediated cytotoxicity). Target cells included the K562 cell line (natural killer cell target), chicken erythrocytes (CRBCs), and herpes simplex virus-infected CEM cell lines. Newborn PMNs were equivalent to adult PMNs in their cytotoxic capacity in several cytotoxicity assays. Neither adult nor newborn PMNs lyse tumor cell targets (i.e., K562 cells) spontaneously, but both lyse K562 cells following activation with phorbol myristate acetate. Both adult and newborn PMNs lyse CRBCs and herpes simplex virus-infected CEM cells in antibody-dependent cellular cytotoxicity assays, and this lysis could be enhanced by the cytokines granulocyte-macrophage colony-stimulating factor and gamma interferon. PMN heteroantibody-mediated cytotoxicity, resulting from the use of an antibody with dual specificity to CRBCs and immunoglobulin G FcRII, was greater in newborn PMNs than in adult PMNs; however, monocyte heteroantibody mediated cytotoxicity, resulting from the use of an antibody to CRBCs and monocyte immunoglobulin G FcRI, was lower in newborn monocytes than in adult monocytes. The percentage, but not the density, of PMNs expressing FcRII was significantly reduced in newborn PMNs compared with that in adult PMNs, while the percentages and densities of FcRI expression were equivalent in newborn and adult monocytes. We conclude that the cytotoxic capability in term newborn PMNs is equivalent to that in adult PMNs, that the activity of newborn PMNs can be enhanced by antibody and/or cytokines, and that PMNs can contribute to the newborn's ability to kill virus-infected cells. PMID- 7496978 TI - Use of zidovudine in human immunodeficiency virus-induced hepatitis. AB - Hepatitis is a common complication of human immunodeficiency virus (HIV) disease in children. Often a distinct pathogen cannot be identified. Therefore, by exclusion, one must consider HIV the direct pathogen. Zidovudine (AZT) has not been used as a treatment for HIV-positive children with presumed HIV-induced hepatitis because of the potential of this medication to exacerbate preexisting hepatitis. We have used AZT to treat an 8-month-old female with severe HIV induced hepatitis and have achieved complete remission with this regimen. For some HIV-positive children with HIV-induced hepatitis, AZT may be the drug of choice to resolve hepatic inflammation caused by this virus. PMID- 7496974 TI - Levels of circulating fibronectin receptor in adult and pediatric patients with human immunodeficiency virus type 1 infection. AB - We found a significant increase in fibronectin receptor (FNR) levels in the sera of adult human immunodeficiency virus type 1 (HIV-1)-infected patients, especially in those with AIDS (1,026.9 +/- 583.9 ng/ml; P < 0.0001). In contrast, AIDS patients with neurologic disorders and HIV-1-seropositive patients showed normal levels of FNR in serum. In addition, HIV-1-infected children showed increased levels of FNR in serum (824.4 +/- 333.5 ng/ml; P = 0.03). We suggest that an increase of FNR levels in AIDS patients is related to enhanced expression of FNR on HIV-1-infected cells. PMID- 7496975 TI - Frequency of biochemical hypothyroidism in sera referred for autoantibody testing. AB - We examined sera submitted for autoantibody testing for thyroid microsome antibodies (TMA), elevated thyroid-stimulating hormone (TSH), and free thyroxine concentrations. The frequency of TMA in antinuclear antibody-positive sera was higher (19%) than that in antinuclear antibody-negative sera (12%). Elevated TSH concentrations in serum and subnormal thyroxine concentrations in serum were associated with the presence of TMA; TMA titer and the frequency of elevated TSH concentrations were also associated with the presence of TMA. PMID- 7496979 TI - Multicenter study of photorefractive keratectomy for myopia of 6.00 to 8.00 diopters. VISX Moderate Myopia Study Group. AB - PURPOSE: To summarize the initial results of excimer laser photorefractive keratectomy (PRK) on 89 eyes of 80 patients with moderate myopia (myopia of -6.00 to -8.00 diopters [D]; mean -6.98 +/- 0.90 D) at nine investigational sites. METHODS: All treatments used an argon fluoride excimer laser (VISX, Inc, Santa Clara, Calif) using standard settings. Sixty eyes received single-zone 6.0 millimeter ablations and 29 eyes received two-zone ablations. Follow up ranged from 1 month (n = 89) to 6 months (n = 46). RESULTS: At 3 months, uncorrected visual acuity measured 20/40 or better in 75% and 20/20 or better in 18%; 78% were within +/- 1.00 D of intended correction, 38% were within +/- 0.50 D, and 9% lost two or more lines of spectacle-corrected visual acuity. At 6 months, uncorrected visual acuity measured 20/40 or better in 74% and 20/20 or better in 17%; 67% were within 1.00 D of intended correction, 38% within 0.50 D, and 2% (1/46) lost two lines of spectacle-corrected visual acuity. CONCLUSIONS: PRK for moderate myopia with large diameter ablation zones appears safe and more predictable than that done using smaller ablation zone diameters. Longer follow up is needed to better define stability, the effects of postoperative corticosteroids, and the use of single-versus double-zone ablations. PMID- 7496981 TI - Locating the corneal sighting center from videokeratography. AB - BACKGROUND: The corneal sighting center is defined by the intersection of the line of sight with the anterior corneal surface and is usually found by measuring angle lambda, between the pupillary axis and the line of sight. It is to be determined if a measurement of the pupil offset in the corneal map of videokeratography might be used instead of angle lambda to provide a practical clinical method for finding the corneal sighting center. METHOD: Two-dimensional angle lambda measurements were made in a group of 17 young adult Caucasian subjects under natural pupil conditions, using an apparatus designed for this purpose, and were compared to measurements of the corneal sighting center position as located by videokeratography. RESULTS: The correlation between angle lambda as measured directly and the pupil offset as measured by videokeratography was 0.84, which is statistically and clinically significant. CONCLUSIONS: The position of the corneal sighting center may be found relative to the corneal reflex of a fixated light by using the distance and direction of the pupil offset in the corneal map of videokeratography, or indirectly by taking 0.6 times the same value and measuring in the opposite direction from the pupillary axis. The latter method does not require active fixation by the patient. PMID- 7496980 TI - Model for deriving the optical performance of the myopic eye corrected with an intracorneal ring. AB - BACKGROUND: The intracorneal ring has been advanced as a mechanical device for the correction of myopia. The device may reduce refractive error, but the effect on the overall optical performance of the eye has been neglected. This paper addresses this issue by presenting mathematical models that will predict the effect of the intracorneal ring on refractive error, corneal asphericity, and the spherical aberration of the eye. MATERIALS AND METHODS: Algorithms are derived for predicting the corneal apical radius, asphericity, and the change in myopia after inserting intracorneal rings of any thickness and diameter. RESULTS: The model predicts that the magnitude of myopia reduction is a function of both ring thickness and diameter. The large diameter (9 to 10 mm), thin (0.1 to 0.2 mm) intracorneal ring is less likely to adversely affect corneal asphericity and therefore not significantly enhance the spherical aberration of the eye. CONCLUSIONS: An intracorneal ring cannot correct more than 4.00 diopters (D) of myopia without significantly increasing ocular spherical aberration, which, in turn, will compromise the final visual outcome. PMID- 7496976 TI - Analysis of tuberculous meningitis cases by an immunoblotting assay based on a mycobacterial antigen complex. AB - Tuberculous meningitis cases were analyzed by an immunoblotting test based on Mycobacterium bovis BCG antigen complex A60. Anti-A60 immunoglobulin G (IgG) in cerebrospinal fluid (CSF) allowed early diagnosis, and concentrations decreased after recovery. In primary meningitis forms, anti-A60 IgGs were intrathecally synthesized and specific oligoclonal IgGs were present in CSF. In meningeal complications of pulmonary tuberculosis, there were matching titers of anti-A60 IgG in blood and CSF (mirror pattern). Correlation between CSF-restricted patterns and CSF pleocytosis was shown. PMID- 7496977 TI - Enriched HLA-DQ3 phenotype and decreased class I major histocompatibility complex antigen expression in recurrent respiratory papillomatosis. AB - Respiratory papillomas, caused by human papillomaviruses, are benign tumors that recur following removal. We evaluated immune function and major histocompatibility complex (MHC) phenotype and expression in these patients. MHC independent immune function appeared normal. The frequency of peripheral blood MHC class II phenotypes was highly enriched for DQ3 and DR11, one split of DR5. Class I MHC antigen expression on papilloma tissue was markedly reduced. Together, these phenomena may facilitate papillomavirus evasion of the cellular immune response. PMID- 7496983 TI - Cognitive styles and personality characteristics strongly influence the decision to have photorefractive keratectomy. AB - BACKGROUND: A substantial number of patients who elect to undergo photorefractive keratectomy do so without the motivation of occupational uncorrected vision requirements. We hypothesized that information processing preferences for the auditory (versus visual) modality in a global, associative (versus detailed, sensory-oriented) style with adaptability and risk-taking (versus predictability) personality characteristics would predominate in patients electing photorefractive keratectomy. METHOD: Seventy-three prospective photorefractive keratectomy patients attended informational sessions. Sixteen occupationally driven patients and one refusal were excluded from the analysis. The 27 patients electing to proceed with surgery were compared with the 29 declining surgery. Personality characteristics and cognitive styles were determined by the Myers Briggs Type Indicator: Abbreviated Version and the Modality Strengths Indicator. RESULTS: Subjects electing surgery showed significantly greater preferences for processing information in the auditory modality and in a global, associative style, with adaptability and risk-taking personality characteristics. Combining the attributes statistically differentiated the two groups. CONCLUSIONS: Specific cognitive styles and personality characteristics strongly influence the choice to pursue photorefractive keratectomy when that choice is not occupationally driven. PMID- 7496982 TI - Corticosteroid suppression of trauma-induced hyaluronan in rabbit cornea and aqueous. AB - BACKGROUND: Corneal and aqueous hyaluronan have recently been shown to react in response to several different types of trauma, including cataract surgery. In order to find ways to influence the reaction, we have evaluated the effect of topical dexamethasone (Isopto-Maxidex, Alcon Universal Ltd, Fort Worth, Tex) or indomethacin (Confortid, Dumex Ltd, Copenhagen, Denmark) on the postoperative hyaluronan concentration in rabbit cornea and aqueous after extracapsular lens extraction. METHODS: The drugs were administered as topical eye drops three times daily. The hyaluronan concentration in rabbit cornea and aqueous after extracapsular lens extraction was measured with a radioligand assay. RESULTS: Dexamethasone treatment (1 mg/mL) significantly suppressed the increase in corneal hyaluronan seen after extracapsular lens extraction, 2 (p < or = 0.0022) and 3 weeks (p < or = 0.0002) after surgery, while indomethacin did not induce any significant difference at 2 weeks. When the dexamethasone concentration was lowered to 0.1 and 0.2 mg/mL, there was still a significant decrease (p < or = 0.009) in hyaluronan concentration, but at lower concentrations of dexamethasone (0.01 and 0.02 mg/mL), no significant decrease was seen. The increase in aqueous hyaluronan concentration seen 2 days after surgery in untreated eyes was significantly lowered by both dexamethasone (p < or = 0.0076) and indomethacin (p < 0.036). CONCLUSIONS: Dexamethasone lowers reactive corneal and aqueous hyaluronan concentration in vivo after extracapsular lens extraction. PMID- 7496984 TI - Successful exchange of dislocated phakic intraocular lens. PMID- 7496986 TI - Informed consent and photorefractive keratectomy. PMID- 7496985 TI - Barraquer Lecture. Refractive surgery: myth versus reality. PMID- 7496987 TI - Mastel Byron Radial Keratotomy Guide. PMID- 7496989 TI - Different regulatory properties of the cytosolic and mitochondrial forms of malic enzyme isolated from human brain. AB - The human brain contains a cytosolic and mitochondrial form of NADP(+)-dependent malic enzyme. To investigate their possible metabolic roles we compared the regulatory properties of these two iso-enzymes. The mitochondrial malic enzyme exhibited a sigmoid substrate saturation curve at low malate concentration which was shifted to the right at both higher pH values and in the presence of low concentration of Mn2+ or Mg2+. Succinate or fumarate increased the activity of the mitochondrial malic enzyme at low malate concentration. Both activators shifted the plot of reaction velocity versus malate concentration to the left, and removed sigmoidicity, but the maximum velocity was unaffected. The activation was associated with a decrease in Hill coefficient from 2.3 to 1.1. The human brain cytosolic malic enzyme displayed a hyperbolic substrate saturation kinetics and no sigmoidicity was detected even at high pH and low malate concentrations. Succinate or fumarate exerted no effect on the enzyme activity. Excess of malate inhibited the oxidative decarboxylation catalysed by cytosolic enzyme at pH 7.0 and below. In contrast, decarboxylation catalysed by mitochondrial malic enzyme, was unaffected by the substrate. These results suggest that under in vivo conditions, cytosolic malic enzyme catalyses both oxidative decarboxylation of malate and reductive carboxylation of pyruvate, whereas the role of mitochondrial enzyme is limited to decarboxylation of malate. One may speculate that in vivo the reaction catalysed by cytosolic malic enzyme supplies dicarboxylic acids (anaplerotic function) for the formation of neurotransmitters, while the mitochondrial enzyme regulates the flux rate via Krebs cycle by disposition of the tricarboxylic acid cycle intermediates (cataplerotic function). PMID- 7496988 TI - Astigmatism and straight transverse corneal incisions. PMID- 7496990 TI - The modulation of the oxidative phase of the pentose phosphate pathway in mouse liver. AB - The glucose-6-phosphate dehydrogenase from mouse liver is fully inhibited in vitro by physiological concentrations of NADPH. This suggests that the oxidative phase of the pentose phosphate pathway requires some deinhibitory system. In order to investigate regulation of the pentose phosphate pathway, various parameters (intermediate concentrations, mass-action ratios of reactions, etc.) were measured in liver from control mice and from meal-fed mice. Assays were also carried out to detect any molecules causing the reverse of glucose-6-phosphate dehydrogenase inhibition by NADPH. The liver of meal-fed mice show greater glucose-6-phosphate dehydrogenase and 6-phosphogluconate dehydrogenase activities. They also had greater concentrations of several metabolic intermediates and triglycerides than the control animals (P < 0.001). These results prove that the diet increases the flow of the pentose phosphate pathway in a lipogenic sense. The glutathione reductase does not change with the diet, suggesting that this enzyme does not participate in the modulating process. Unlike rat liver, no molecules causing the reverse of glucose-6-phosphate dehydrogenase inhibition by NADPH were detected. These data suggest that the increase of flow of the pentose phosphate pathway during lipogenesis is obtained by an increase in enzyme synthesis. PMID- 7496991 TI - Alveolar surfactant subfractions differ in their lipid composition. AB - Alveolar surfactant consists of subfractions which are generated during normal lung function. Although subfractions obtained by differential centrifugation of lung lavage differ in structure, function and protein content, the phospholipid pattern shows only minor differences. To correlate possible differences in composition between subfractions to their functional properties we did a more detailed analysis of lipid pattern. Subfractions of lung lavages from Wistar rats were obtained by differential centrifugation, lipid classes were separated by thin layer chromatography (TLC). Fatty acids and plasmalogens were determined as methylester and dimethylacetals by gas chromatography, respectively. Cholesterol and vitamin E were determined enzymatically and by HPLC, respectively. The patterns of fatty acids of total lipids and of the molecular species of phosphatidylcholine and phosphatidylethanolamine were very similar among the subfractions. The distribution of individual lavage lipids varied considerably. Three types of subfractions can be distinguished: The two dense subfractions (1000 g and 60,000 g) contain 70-88% of total phospholipids, dipalmitoylphosphatidylcholine, polyunsaturated phospholipids and polyunsaturated fatty acids present in lung lavage. The less dense subfraction (100,000 g) contains 44-60% of total cholesterol, choline plasmalogen, ethanolamine plasmalogen and vitamin E. The 100,000 g supernatant contains 40-50% of total tri , diacylglycerols and free fatty acids. Our results support the concept that the 1000 g subfraction contains freshly secreted surfactant. The 60,000 g subfraction likely contains the monolayer and freshly secreted surfactant. The 100,000 g pellet probably contains material "squeezed out" from the monolayer at expiration. Most likely, the supernatant contains material destined for removal from the airspace. PMID- 7496992 TI - Mammalian secreted and cytosolic phospholipase A2 show different specificities for phospholipid molecular species. AB - Previous studies using phospholipid vesicles containing single molecular species have shown cytosolic phospholipase (85 kDa) (PL) A2 to possess a marked preference for arachidonic acid (20:4n-6)-containing species, while secreted PLA2 (14 kDa) exhibited little acyl chain selectivity. In this study, we have defined the molecular specificity of cytosolic PLA2 using phospholipid vesicles derived from rat liver which contain complex mixtures of molecular species. Phosphatidylcholine (PC) and phosphatidylethanolamine (PE) were isolated from rat liver by chloroform and methanol extraction, and solid-phase separation. PC and PE vesicles were hydrolysed by either human recombinant cytosolic or porcine pancreatic PLA2. Molecular species compositions were determined by reverse phase high performance liquid chromatography (HPLC) with post-column fluorescence derivitisation. HPLC analysis after limited hydrolysis demonstrated that the secreted phospholipase A2 showed no significant acyl chain specificity using these phospholipid mixtures. However, the cytosolic enzyme demonstrated a high degree of preference for arachidonic acid-containing species such that there was no hydrolysis of other molecular species. The extent of hydrolysis of PC16:0/20:4 was 1.4-fold greater (P < 0.05, n = 3) than PC18:0/20:4, while PE16:0/20:4 and PE18:0/20:4 were hydrolysed to a similar degree. Under these assay conditions, the cytosolic enzyme showed a preference for PE as compared with PC. This study confirms that cytosolic PLA2 is highly selective for sn-2 20:4n-6-containing phospholipid molecular species even when presented with a complex natural species mixture. This specificity is consistent with the cytosolic enzyme having a primary role in the process of arachidonic release within cells.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7496993 TI - Analysis by limited proteolysis of domain organization and GSH-site arrangement of bacterial glutathione transferase B1-1. AB - Limited proteolysis method has been used to study the structure-function relationship of bacterial glutathione transferase (GSTB1-1). In absence of three dimensional structural data of prokaryote GST, the results represent the first information concerning the G-site and domains organization of GSTB1-1. The tryptic cleavages occur mainly at the peptide bonds Lys35-Lys36 and Phe43-Leu44, generating two major molecular species of 20-kDa, 3-kDa and traces of 10-kDa. 1 chloro-2,4-dinitrobenzene favoured the proteolysis of the 20-kDa fragment markedly enhancing the production of the 10-kDa peptide by cleaving the chemical bonds Lys87-Ala88 and Arg91-Tyr92. The tryptic cleavage sites of GSTB1-1 was found to be located close to those previously found for the mammalian GSTP1-1 isozyme. It was concluded that despite their low sequence homology (18%), GSTB1-1 and GSTP1-1 displayed similar structural features in their G-site regions and probably a common organization in structural domains. PMID- 7496995 TI - Hyperosmolality stimulates phospholipase A2 activity in rabbit renal medulla and in Madin-Darby canine kidney (MDCK) cells. AB - Renal medullary cells are able to accumulate glycerophosphocholine during adaptation to the high extracellular osmolality. The aim of this study was to investigate the effect of hyperosmolality on both phospholipase A2 activity and the rate of choline incorporation into glycerophosphocholine in rabbit renal medulla and Madin-Darby Canine Kidney cells. Phospholipase A2 activity was assayed in cellular subfractions isolated from both rabbit kidney medulla and Madin-Darby Canine Kidney cells in the presence of either 1-palmitoyl-2-[1 14C]palmitoyl phosphatidylcholine or 1-stearoyl-2-[1-14C]arachidonyl phosphatidylcholine as substrate. The rate of choline incorporation into glycerolphosphocholine was measured in Madin-Darby Canine Kidney cells growing in the presence of [methyl-3H]choline in the growth medium. Water deprivation of rabbits resulted in an increase of phospholipase A2 activity from 2.7 +/- 0.4 (n = 5) and 5.7 +/- 0.7 (n = 5) to 5.0 +/- 0.8 (n = 5) and 10.8 +/- 1.3 (n = 5) pmol of fatty acid released/min per mg protein in mitochondrial and microsomal fractions, respectively, using dipalmitoyl phosphatidilcholine as substrate while the activity of cytosolic enzyme remained unchanged. Similarly, the addition of sodium chloride in order to increase growth medium osmolality (from 320 mOsm/kg to 520 mOsm/kg) resulted in an elevation of both mitochondrial (from 1.8 +/- 0.1 to 4.9 +/- 0.8 pmol of fatty acid released/min per mg protein, (n = 4) and microsomal (from 8.7 +/- 0.5 to 15.9 +/- 1.7 pmol of fatty acid released/min per mg protein, n = 4) phospholipase A2 activities.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7496994 TI - Nucleoside phosphatase activities on pig pancreas zymogen granule membranes analyzed by non-denaturing polyacrylamide gel electrophoresis. AB - The membrane of the pancreatic zymogen granule plays an important part in the sequence of storage, transport and exocytosis of digestive enzymes. While much is known on stimulus-secretion coupling, very little is understood about how the storage organelles move in the cytoplasm to the luminal plasma membrane and why and how they fuse with it to release the contents. It is assumed that nucleoside phosphatases are involved in these energy consuming processes. Pancreatic zymogen granule membranes contain one major glycoprotein, GP-2, and a few minor proteins all with unknown functions. In order to identify functions we have purified zymogen granule membranes from pig pancreas, solubilized the proteins under non denaturing conditions with the detergent CHAPS and characterized the extracted proteins by polyacrylamide gel electrophoresis, histochemistry and lectins. Three major protein bands, often fused in one broad band, revealed enzymatic activity for adenosine-, cytidine-, inositol- and guanidine- di- and triphosphates by the precipitation of liberated phosphate by Pb(NO3)2. This activity was sensitive to known ATP diphosphohydrolase inhibitors. The band with activity arises from a 92 kDa glycoprotein. A different narrow band showed monophosphatase activity for AMP, GMP, IMP and CMP. Some of the activities were inhibited by different lectins, indicating glycosyl groups near the active site. Electron microscopical cytochemistry confirmed a nucleoside phosphatase activity on granule membranes. Our results show for the first time that the nucleoside phosphatase activity of the zymogen granule membranes is carried by a 92 kDa glycoprotein, probably the known self-associating form of GP-2. The hydrolysis of tri- and diphosphate nucleotides could provide the energy required by exocytosis. PMID- 7496997 TI - The dephosphorylation of AMP and IMP by a soluble low Km 5'nucleotidase from human seminal plasma: some regulatory aspects. AB - In this study, a soluble low Km 5'nucleotidase, dephosphorylating IMP with a Vmax/Km ratio 10-times higher than that of AMP, has been purified from human seminal plasma. The effect of inorganic phosphate (Pi) and adenylate energy charge variations on the activity of this enzyme has also been investigated. In the physiological range, with IMP as substrate, the activity of the enzyme does not change whereas the hydrolysis of AMP increases with decreasing energy charge values. In the presence of both the substrates, phosphate exerts an inhibitory effect on the enzyme activity with a similar concentration dependence pattern. The results show that AMP-hydrolysing activity responds to variations of energy charge by increasing the AMP degradation thus protecting the value of energy charge at the expense of a decrease in the total adenylate pool. In contrast, the dephosphorylation of IMP is not regulated by changes in energy charge. This data suggests that the degradation of IMP and AMP, although carried out by the same enzyme, is controlled by different regulatory mechanisms. PMID- 7496996 TI - ADP-ribosylation in adrenal glands: purification and characterization of mono-ADP ribosyltransferases and ADP-ribosylhydrolase affecting cytoskeletal actin. AB - Mono-ADP-ribosylation in mammals is poorly understood. In this study, we purified four mono-ADP-ribosyltransferases and one ADP-ribosylhydrolase from rat adrenal medulla. The four purified mono-ADP-ribosyltransferases had molecular weights of 69,000 by gel filtration, pH optima of 8.0, and Kms for their action on NAD of about 20 microM. The four enzymes ADP-ribosylated to the alpha-subunit of heteromeric GTP-binding proteins. After tryptic digestion of alkylated actin mono ADP-ribosylated by the purified mono-ADP-ribosyltransferases or botulinum C2 toxin, the two radioactive peptide patterns were identical. The purified ADP ribosylhydrolase with mono-ADP-ribosylated actin as the substrate had a molecular weight of 61,000 on gel filtration, a pH optimum of 7.5, and a Km for mono-ADP ribosylated actin of about 7 microM. The enzyme released ADP-ribose from ADP ribosylated actin and the alpha-subunit of hetromeric GTP-binding proteins. Actin monomers mono-ADP-ribosylated by the four mono-ADP-ribosyltransferases did not form actin filaments after the addition of Mg2+. After release of ADP-ribose by ADP-ribosylhydrolase, actin filaments formed on the addition of Mg2+, suggesting that the polymerization and depolymerization of cytoplasmic actin the adrenal chromaffin cells may be regulated by mono-ADP-ribosylation. PMID- 7496999 TI - Different influences of two fractions of lung cytochrome b5 on reconstituted lung benzphetamine N-demethylase system. AB - Chromatography of lung microsomal cytochrome b5 obtained from DEAE-cellulose columns, yielded two distinct cytochrome b5 fractions. These cytochrome b5 fractions were further purified by Sephadex G-100 gel filtration chromatography. The specific cytochrome b5 content of fraction 1 and fraction 2 was found to be 16.5 and 16.4 nmol/mg protein respectively. Both fractions were free of cytochrome P-450, NADPH-cytochrome P-450 reductase and NADH-cytochrome b5 reductase activities. The effects of lung cytochrome b5 (fraction 1 and fraction 2) and liver cytochrome b5 on benzphetamine N-demethylase activity were examined. Four different reconstitution systems were used. Lung cytochrome b5 fraction 2 and liver cytochrome b5 stimulated N-demethylase activity in all four systems when b5:P-450 molar ratio was low, i.e. 0.25 or 0.5. Both cytochrome b5 samples inhibited N-demethylase activity when b5:P-450 ratio exceeded 1:1 molar ratio. In contrast lung cytochrome b5 fraction 1 stimulated N-demethylase activity in all four systems. Maximal enhancement of the activity was obtained when b5:P-450 ratio was 0.5. The greatest increase in N-demethylation activity was observed in the reconstitution system with the lowest concentration of cytochrome P-450 reductase, conditions which most closely resemble intact microsomes. PMID- 7496998 TI - Aminoacid utilization by Helicobacter pylori. AB - Utilization of aminoacids during growth by laboratory adapted and wild type Helicobacter pylori strains was investigated employing nuclear magnetic resonance spectroscopy and aminoacid analysis. All H. pylori strains tested showed growth rates with doubling times of approx. 11.5 hr in liquid cultures with semi-defined media or with defined aminoacid broth without carbohydrates. Fast utilization of several aminoacids at rates between 80 and 250 microM/hr was observed in culture broths inoculated with approx. 10(7) cells/ml; and acetate, formate and succinate accumulated as catabolic products in the growth media. Suspensions of bacterial cells and lysates in isotonic solutions converted arginine, asparagine, aspartate, glutamine, and serine used as sole substrates at significant rates; and under these conditions the principal metabolic products observed were acetate, formate, succinate and lactate. The findings of the study indicated that H. pylori can survive employing aminoacids as the basic nutrients, and suggested some of these metabolites were utilized via fermentative pathways with common characteristics to those found in anaerobes. PMID- 7497000 TI - The regulation of IGF-I receptor gene expression. AB - The insulin-like growth factor I (IGF-I) receptor mediates most of the biological effects of IGF-I and -II. Despite its structural similarity to the insulin receptor, the IGF-I receptor is mainly involved in the transduction of growth and differentiation types of signals. The IGF-I receptor gene is constitutively expressed by most cells in the organism as well as in culture, consistent with the role of the IGFs as survival factors. In addition, the expression of the IGF I receptor gene is modulated by a number of physiological and pathological factors, including developmental stage, nutritional status, hormones, growth disorders and malignancy. The regulatory region of the IGF-I receptor gene has been characterized and shown to display a high level of basal promoter activity. Transcription factor Sp1 is a strong activator of IGF-I receptor gene expression, whereas tumor suppressor WT1 represses its activity. The biological implications of these findings in both normal development and disease are described in this review. PMID- 7497001 TI - Phospholipid composition, phospholipase A2 and sphingomyelinase activities in rat liver nuclear membrane and matrix. AB - Little data has been published regarding the presence of lipolytic--neutral sphingomyelinase and phospholipase A2--activities in the nuclear matrix and membranes of cells. Considering the influence of phospholipids and the above enzymes on transcription and replication, the phospholipid composition and lipolytic activities of the nuclear matrix and membrane was determined. Pure nuclei from normal rat liver cells were isolated after nuclease digestion and extraction of the nuclear material with low and high ionic strengths, in presence of reducing agents. Phosphatidylcholine was the main phospholipid in the nuclear membranes (44% from the total phospholipids) whereas the amount of phosphatidylserine was highest in the nuclear matrix (45%). The specific activity of phospholipase A2 in nuclear membranes was similar to that from liver plasma membranes and in hen erythrocyte nuclear membranes, (2.3 +/- 0.6 nmol/mg/min, SEM, n = 8) while in the nuclear matrix it was 20 times higher. It did not show specificity towards phosphatidylcholine and phosphatidylethanolamine as substrates. A high sphingomyelinase activity in the nuclear matrix of rat liver was found (12.7 +/- 2.1 nmol/mg/min, SEM, n = 8) and this activity was affected by reducing agents (dithiotreitol). Our results showed that phospholipids phosphatidylcholine and phosphatidylserine were constituent part of the nuclear structures as were the enzymes phospholipase A2 and sphingomyelinase. This is consist with a role for lipids and their breakdown products in metabolic events within the nucleus. PMID- 7497002 TI - Who supervises the students? PMID- 7497003 TI - Mothers, daughters and death. PMID- 7497004 TI - Tuberculosis: a risk to nurses. PMID- 7497007 TI - Raising nurses' awareness. PMID- 7497010 TI - A nurse challenges patriarchy. PMID- 7497006 TI - Controlling infection. PMID- 7497008 TI - Further directions for learning. PMID- 7497009 TI - Is nursing a crime? PMID- 7497005 TI - Manslaughter and the case for nurses. PMID- 7497011 TI - Daring to question. PMID- 7497012 TI - [Early amniocentesis--already an alternative?]. PMID- 7497013 TI - [Monitoring the pregnant diabetic patient]. AB - Despite improved surveillance of pregnant diabetic women perinatal mortality and morbidity remains higher than in the general population. Low detection rates of patients with gestational diabetes represent one of the main reasons as screening programs based upon the presence of risk factors only comprise 30% of all women with gestational diabetes. Concerning maternal risks in patients with insulin dependent diabetes mellitus the incidence of pregnancy induced hypertension is increased up to 12-28%. Macrosomia (6-32%) and malformations (1.5-6%) are the most frequent fetal complications and depend on the quality of controlling the blood glucose level. However, the decrease of fetal and maternal risks requires a general screening program for gestational diabetes as well as an intensive surveillance of the mother and the fetus by an obstetrician and internal specialist, respectively. Delivery of pregnant diabetic women should preferable be performed in specialized hospital units. PMID- 7497014 TI - [Diabetes screening in pregnant patients with different risk factors. Benefits, therapeutic success, consequences]. AB - 600 ethnically heterogeneous patients with various risk factors from outpatient obstetric clinic of the UFK Ulm were screened for gestational diabetes. Incidence with the 100 g oGTT was 21%, with fasting levels having the lowest prognostic value, and one hour levels having the best. After the diagnosis of gestational diabetes was established patients were counselled and put on diet, 7% received insulin additionally. Weight gain was far more acceptable in those patients compared to the group of patients with normal metabolism. We observed no significant differences concerning fetal macrosomia, caesarean section-rates, newborn apgar and ph-values compared to normal pregnant women. We confirmed the known risk factors, personal history, familial disposition and maternal age. A screening based only on these criteria, however, would fail to detect 20% of patients with gestational diabetes. A general screening must be postulated. PMID- 7497015 TI - [Estriol and human placental lactogen as risk indicators in pregnancies of diabetic and hypertensive patients]. AB - The correlation between estriol (E3) and human placenta lactogen (HPL) and perinatal morbidity was investigated in 105 diabetic, 96 hypertensive and 96 pregnancies without diabetes and hypertension. The hormones were determined by radioimmunoassay. Only if two determinations were outside the normal range the values were accepted as pathological. In pregnancies with hypertension a decrease of E3- and HPL-concentrations was correlated to significant higher incidence of operative deliveries and pathological APGAR-Scores. In the diabetic group is the low E3-concentration of significant predictive value in respect of the pathological cardiotocography as the indication of the operative delivery, the same applies to low HPL-concentrations in the hypertension group, in the control group no significant differences could be determined relating thereto. Hypotrophic newborns in the hypertension and control group are significant more frequent, if the concentration of E3 is decreased. The same applies to decreased HPL-concentrations in every group, where the hypertension group shows the highest predictive values. PMID- 7497016 TI - [Influence of non-diabetic pregnancy on fructosamine and HbA1c concentration]. AB - HbAIc and fructosamine concentrations were measured in the course of 177 nondiabetic pregnancies and compared with the corresponding values of 24 nondiabetic nonpregnant women. In all three trimesters HbAIc and fructosamine were significantly lower than the corresponding values in the nonpregnant women; HbAIc: 1st trimester 4.77 +/- 0.62%, 2nd trimester 4.38 +/- 0.59%, 3rd trimester 4.33 +/- 0.49%, p < 0.01; fructosamine 1st trimester 2.13 +/- 0.17 mmol/l, 2nd trimester 2.02 +/- 0.15 mmol/l, 3rd trimester 1.90 +/- 0.15 mmol/l, p < 0.01; nonpregnant women: HbAIc 5.13 +/- 0.41%, fructosamine 2.53 +/- 0.17 mmol/l. However, if the fructosamine is correlated to the respective total protein concentration a constant value results for the course of pregnancy. The changes in the HbAIc and fructosamine concentrations in pregnancy should be taken into account when treating pregnant diabetics. PMID- 7497017 TI - [Risk groups of newborn infants of diabetic mothers in relation to their somatic outcome and maternal diabetic metabolic status in pregnancy]. AB - In a prospective study, neonatal morbidity of newborn children of diabetic mothers and its association with the maternal metabolism was determined. Particular attention was directed on the somatic outcome of the children and their frequent metabolic imbalances. In addition, we determined the influence of maternal biological and somatometrical variables on the somatic outcome of newborns. Dependent upon the mothers' and children' variables, risk groups of newborns (fetopathy groups) were defined to optimize clinical care and surveillance of newborns. A total of 810 children were included born to mothers with primary insulin dependent diabetes mellitus (IDDM), non insulin dependent diabetes (NIDDM), or gestational diabetes (GDM). Among the study population, 41.7% of children had macrosomia, 27.2% had a weight-length index > 1.2, 17.9% developed hypoglycemia and 19.5% hyperbilirubinemia within the initial 72 hours after birth. The somatic outcome of the children was significantly associated with pregnancy duration, maternal age, weight, height, and HbA1. Increasing maternal HbA1 prior to delivery (categorized in < 8.5%, 8.6-10%, > 10%) was associated with increased relative risk of incidence of neonatal morbidity. Finally, risk groups (fetopathy groups I-III) were defined according to maternal HbA1 value and somatic outcome of the newborns. The importance of these fetopathy groups for criteria of neonatal morbidity is demonstrated. Based upon categorization of newborn children into fetopathy groups, children should be allocated to specific concepts of appropriate surveillance and clinical care. The fetopathy classification may also serve as an independent tool for retrospective quality control of diabetic pregnancy. PMID- 7497019 TI - [Detecting fetal hypoxemia sub partu]. PMID- 7497018 TI - [The course of pregnancy and fetal outcome in diabetic patients with anamnestic fetal death]. AB - Of 116 pregnant patients with diabetes mellitus type 1 or gestational diabetes, 12 (10.3%) had previously suffered stillbirth, while in four (3.4%) cases spontaneous abortion after the 6th month had occurred. In these 16 cases, we examined the mean daily blood glucose concentration, glycosylated hemoglobin (HbA1c), fructosamine as well as glucose and albumin elimination via urine. These control data on carbohydrate metabolism were correlated with the intensity of gestational monitoring in relation to course of pregnancy and neonatal morbidity and/or mortality. In addition, we retrospectively analyzed the cause of prior intrauterine death. In six of eight cases evaluable, we could hereby demonstrate disturbed carbohydrate metabolism. Pregnant women with diabetes mellitus type 1 were hospitalized twice as often (on average four times) as patients with gestational diabetes (2.1 admissions on average). While five of seven type 1 diabetics were primarily admitted prior to the 20th week of gestation, the majority of gestational diabetics (7/9) were first examined by qualified medical personnel trained in the care of gestational diabetes after the 28th week of gestation. Two of nine were first diagnosed post partum. 13 of 14 patients with prepartally known diabetes demonstrated mean blood glucose values under 110 mg/dl. While one of the two patients with postpartally diagnosed diabetes suffered stillbirth once again, the other had to be delivered by emergency C section because of fetal distress (pH art. 7.08). Our results show that adequate monitoring of carbohydrate metabolism and proper care of diabetic pregnant women can lower obstetrical/neonatal risks to more acceptable levels, especially in those patients who have had stillbirth in their prior medical history. PMID- 7497020 TI - [Pre- and postpartum management of hypoplastic left heart syndrome]. PMID- 7497021 TI - [Comment on F. Stoz et al.: is general amalgam prohibition justified?]. PMID- 7497022 TI - [Aged couples and management of dependence. Study of 75 helping spouses]. AB - The article displays the findings of a survey conducted among 34 husbands and 41 wives who are helping their disabled spouses. The questionnaire deals with the degree of dependency of the helped spouse, his/her health status, the health status of the helper, how the latter copes with the helping tasks, how these tasks are felt and carried out, how is the relationship between the spouses in the past and at present, what are the prospects envisaged by the helper regarding in particular the transfer of the disabled spouse to a nursing home. A scale is used for measuring the various answers. PMID- 7497023 TI - Prepaid financing of primary health care in Guinea-Bissau: an assessment of 18 village health posts. PMID- 7497024 TI - [Medical beliefs and practices in Algeria. Study of various therapeutic routes]. AB - In Algeria, the official health policy is based on a constant principle: the implementation of a health care system patterned on the Western model. As the country is rooted in a non-Western tradition, two paradigms are competing. The article is aimed at measuring the degree of adhesion given to each paradigm (and system of reference) by the people. In short its objective is to determine how a patient believes respectively in the "western" medicine and the "traditional" on and how he accepts they coexist for alleviating his sufferings. In a society predominantly religious and rural, the therapeutic relation is evaluated firstly on the basis of the kindness and warmth of the care providers. It is also appreciated by its religious surroundings. These two elements are absent in Western-style hospitals. Other factors contribute to strengthen the traditional medicine among the people. PMID- 7497025 TI - [Contraception and abortion in the countries of eastern Europe]. AB - For various reasons, including the population policies of the former Socialist countries, abortion is much more frequent than contraception in Central and Eastern Europe. The contrary occurs in Western countries. In the current context, countries in Central and Eastern Europe should improve abortion services and this should be a priority target in their health policy. The study does not detect a clear-cut advantage as concerns substituting contraception to abortion in these countries in terms for instance of reducing maternal mortality. In fact a health policy favouring contraception instead of abortion is firstly inspired by political or ethical reasons. If implemented in Central and Eastern Europe, it would require several decades to bear evident results. PMID- 7497026 TI - Smoking cessation efforts. PMID- 7497027 TI - Point-of-care blood testing and cost-effective emergency medicine. PMID- 7497028 TI - Smoking habits of emergency department patients: an opportunity for disease prevention. AB - OBJECTIVE: To determine smoking habits, levels of addiction, readiness to quit, and access to primary care among ED patients. METHODS: A questionnaire was administered prospectively to all noncritical adult patients who presented to one university hospital ED during 23 randomly selected four-hour time blocks; 336 (89%) of 376 eligible patients responded. Self-reported smoking was validated by carbon monoxide breath testing in a pilot sample of 49 patients. RESULTS: The study patients were mostly young (mean age = 35 +/- 15 years), female (59%), white (62%), and high school-educated (73%). Of the 336 ED patients, 41% were current smokers (95% CI = 0.36-0.46); 42% of these were "moderately" to "very highly" dependent on nicotine (Fagerstrom Test for Nicotine Dependence > 4). Of those who smoked, 68% stated they wanted to quit, and 49% wanted to quit within the month. Fifty-six percent of all those who smoked stated that they had never been told to quit smoking by any physician. Thirty-five percent of the ED sample (118 patients) relied upon EDs for most or all of their routine, primary health care; 55% (95% CI = 0.46-0.64) of these patients were current smokers. CONCLUSIONS: The prevalence rates of smoking and nicotine addiction among ED patients are high. Almost half of ED smokers are ready to quit, but most state they have never been told by a physician to do so. Finally, a large proportion of ED smokers receive their primary care in EDs. Therefore, the ED may be an underused setting for smoking cessation intervention. PMID- 7497029 TI - Evaluation of a portable clinical blood analyzer in the emergency department. AB - OBJECTIVE: To assess the potential effects of rapid bedside blood analysis on patient management in the ED. METHODS: A prospective, nonrandomized clinical study was conducted over a consecutive ten-month period (August 1992 to May 1993). Blood samples drawn from a convenience sample of 960 patients for analysis of Na+, K+, Cl-, BUN, glucose, and/or hematocrit (Hct) were simultaneously analyzed by portable clinical analyzer (PCA) and by routine methods in the central laboratory. Caregivers were blinded to the PCA values; patient care was based solely on central laboratory results. Physicians were surveyed after the completion of patient care. RESULTS: The PCA results were available 31 minutes (mean) sooner than were the central laboratory results for Hct, 43 minutes faster for Na+, K+, and Cl-, and 44 minutes faster for BUN and glucose. Except for Hct and glucose, the values obtained from the PCA were not significantly different from the central clinical blood analyzer laboratory values. When surveyed, the physicians caring for the patients reported that had the PCA results been available, a different or an earlier therapeutic approach would have resulted in 9.5% of the cases. The decision to release or admit the patient was based on one or more of the laboratory values for 10.7% of patients sampled. In no case in this series did a physician report that final ED clinical outcome would have been affected. CONCLUSIONS: In our ED, the PCA yielded faster reporting of laboratory values. These earlier results might have reduced the length of stay in the ED for 17.3% of patients studied. Selective use of a handheld portable analyzer might decrease time to therapeutic interventions and time to disposition. PMID- 7497030 TI - Cocaine-associated chest pain: one-year follow-up. AB - OBJECTIVE: To determine the one-year mortality and incidence of myocardial infarction (MI) post-hospital discharge or ED release for patients with cocaine associated chest pain. METHODS: A prospective, observational study of an inception cohort of consecutive patients who presented to one of four municipal hospital EDs with cocaine-associated chest pain. Patients were followed for one year from the end of the enrollment period. Main outcome parameters were the one year actuarial survival and the frequency of nonfatal MI. RESULTS: Mortality data were available for all 203 patients at a mean of 408 days. Additional clinical information was available for 185 patients (91%). There were six deaths (one-year actuarial survival 98%; 95% CI, 95-100%); none from MI. Nonfatal MI occurred in two patients (1%; 95% CI, 0-2%). Continued cocaine use was common (60%; 95% CI, 52-68%) and was associated with recurrent chest pain (75% vs 31%, p < 0.0001). No MI or death was reported for patients who claimed to have ceased cocaine use. CONCLUSIONS: Patients who presented with cocaine-associated chest pain commonly continued to use cocaine after discharge. Urgent evaluation of coronary anatomy or cardiac stress tests may not be necessary for patients for whom MI is ruled out and who do not have recurrent potentially ischemic pain. The subsequent risk for MI and death in this group appears to be low. Intervention strategies should emphasize cessation of cocaine use. PMID- 7497032 TI - Utility of routine admission serum chemistry panels in adult trauma patients. AB - OBJECTIVES: To determine the frequency of abnormal serum chemistries and whether they provide clinically useful information regarding trauma patients. To identify clinical criteria associated with critical serum chemistry values (CSCVs). METHODS: The records of all trauma patients admitted to one urban, Level I Trauma Center were retrospectively reviewed for the period (July 1-December 31, 1989). All trauma patients who had had serum chemistry determinations at hospital admission and at least once more prior to discharge were studied. The CSCVs were determined by a panel of experts prior to record review. Serum chemistry values, patient demographics, mechanisms of injury, and outcomes were statistically analyzed. RESULTS: Most (814/913; 89%) trauma patients had abnormal serum chemistry values. However, only 54/913 (6%) had CSCVs and only six of these CSCVs had prompted a change in resuscitation or treatment (therapeutic K+ infusions). Age > 50 years, a history of hypertension therapy, and a Glasgow Coma Scale score < or = 10 were associated with CSCVs; and CSCVs were correlated with increased mortality and critical care unit admission. CONCLUSION: Routine-admission serum chemistry values, while frequently abnormal for major trauma patients, generally do not provide clinically useful information in the resuscitation and treatment of trauma patients. Hypoglycemia and metabolic acidosis were more rapidly determined using bedside glucose determination and arterial blood gas evaluation. The routine-admission serum chemistry panel described in the study lacked utility for most trauma patients. Selective chemistry panel ordering should be used at the time of hospital admission for major trauma patients. PMID- 7497031 TI - Ethanol-associated hypoglycemia is uncommon. AB - OBJECTIVE: To determine the association of ethanol intoxication with hypoglycemia in ED patients. METHODS: Retrospective, laboratory log review of 953 consecutive patients who were evaluated for ethanol intoxication in an urban university hospital ED over a three-month period. Simultaneous serum glucose determination was carried out for each patient and associations between ethanol level and glucose were sought. RESULTS: Glucose concentrations were unavailable for 16 patients (1.7%). Of the remaining patients, 584 patients had detectable ethanol concentrations (ethanol-positive), and 353 had no detectable ethanol (ethanol negative). Ethanol concentrations (mean +/- SD) in the ethanol-positive group were 50.11 +/- 24.08 mmol/L (231 +/- 111 mg/dL), and glucose concentrations were 5.83 +/- 1.94 mmol/L (105 +/- 35 mg/dL). Hypoglycemia [glucose < 3.72 mmol/L (67 mg/dL)] was observed for five (0.9%) ethanol-positive patients. It was classified as mild-moderate [2.78-3.66 mmol/L (50-66 mg/dL)] for four patients (0.7%) and severe [< 2.78 mmol/L (50 mg/dL)] for one (0.2%). Ethanol concentrations ranged from 25.60 to 68.33 mmol/L (118 to 315 mg/dL). There was no correlation between ethanol and glucose concentrations in any subset of the ethanol-positive patients population. In the ethanol-positive group, patients who had several ethanol positive visits (56 patients, mean 3.6 visits/patient) accounted for four of the five episodes of hypoglycemia, including the one episode of severe hypoglycemia. The frequency of hypoglycemia in repeat visitors (2.0%) was higher than that in the group of patients without repeat visits (0.2%). In the ethanol-negative group, there were four (1.1%) episodes of mild-moderate hypoglycemia and no severely hypoglycemic patients. Hypoglycemia was not more likely to occur among ethanol-positive patients than it was among ethanol-negative patients. CONCLUSION: Hypoglycemia was uncommonly associated with ethanol intoxication, and was found almost exclusively among patients with several ethanol-positive visits. Glucose and ethanol concentrations do not show any linear correlation; patients with higher ethanol concentrations are not at higher risk of hypoglycemia. Hypoglycemia is not more likely to occur in ethanol-positive than in ethanol negative patients. Initial glucose screening does not appear to be necessary for all patients suspected of intoxication; selective screening may be more appropriate. PMID- 7497034 TI - Injuries resulting from bicycle collisions. AB - OBJECTIVE: To review all bicycle crash-related injuries reported to the Oregon Injury Registry for 1989 to compare patterns of injury and other features in adults vs children and adolescents. METHODS: A retrospective descriptive study was conducted using data from the Oregon Injury Registry. For 1989, this registry included all injury-related deaths and approximately 75% of hospital admissions of 24 hours or more for injury in the State of Oregon. Deaths, helmet use, neurologic injuries, and concurrent ethanol use were evaluated for all patients and for the two age groups. RESULTS: There were 311 bicycle-related injured patients in the registry for 1989; 122 (40%) were adults (age > or = 21 years) and 189 (60%) were children/adolescents (age < 21 years). Approximately 69% of both age groups were male. All of the 15 deaths involved male patients and most deaths [10/15 (67%)] involved injured adults. Bicycle vs motor vehicle collisions accounted for 14 (93%) deaths and 106 (34%) of all registry entries. While only 19 (15%) of the injured adults had elevated blood alcohol levels, half the adults who died had been intoxicated. Helmet use was rare with only 12 (4%) of all the injured riders known to have been helmeted; no rider who died was known to have been helmeted. Neurologic injuries were common. In children, 27 (14%) had sustained skull fractures, 36 (19%) intracranial injuries, and one (0.5%) a spinal injury. In adults, 13 (10%) had sustained skull fractures, 32 (26%) intracranial injuries, and three (2%) spinal injuries. CONCLUSIONS: Although children account for 60% of the serious bicycle injuries in Oregon, adults account for 67% of the deaths. Helmet use is rare, brain injuries are frequent, and alcohol use appears to be a contributing factor in cycling deaths among adults. Public education efforts should be directed to both adult and pediatric populations, emphasizing safe cycling practices and helmet use. PMID- 7497033 TI - Noninvasive evaluation of the collateral circulation to the hand. AB - OBJECTIVE: To compare four methods of evaluating collateral blood flow to the hand. METHODS: The hands of 74 volunteers on the faculty or staff of a university hospital were studied prospectively. Only subjects without known peripheral vascular disease were included. Four tests were used in random order to assess radial and ulnar artery flow. Results of the assessments using the modified Allen's test, pulse oximetry, plethysmography, and laser Doppler perfusion monitoring were compared. RESULTS: No interrupted palmar arch was found. The modified Allen's test was normal in all cases. Pulse oximetry detected a 5% incidence of noticeably reduced blood flow in one artery compared with the other artery. This dominance of one artery was identified in 69% of the hands by plethysmography. The laser Doppler noted a dominant artery in 64% of the hands. Plethysmography and the laser Doppler disagreed in their findings in only 9% of the hands evaluated. Numerical values of blood flow, attainable only by the laser Doppler, were significantly lower upon occlusion of the radial vs the ulnar arteries (p < 0.05; paired t-test). CONCLUSION: All of the tests provide information about the collateral circulation to the hand. Only the laser Doppler provides quantitative blood flow. Further studies involving subjects most at risk for post-cannulation ischemic injury are needed to guide the clinical application of these findings. PMID- 7497035 TI - Artificial surfactant administration in an animal model of near drowning. AB - OBJECTIVE: To study the use of exogenous surfactant in a rabbit fresh-water near drowning model. METHODS: In a randomized, placebo-controlled experiment, 21 rabbits were anesthetized, paralyzed, and subjected to near drowning with 6 mL/kg of distilled water. Vital signs, arterial blood gases, and pulmonary compliance were measured at predetermined intervals. The animals were randomized into two groups: a normal O2 group that had their pO2 levels adjusted to 75-150 torr using supplemental O2 as needed, and a high O2 group ventilated with an FIO2 = 100%. Ventilator settings were further adjusted to normalize pH and pCO2 levels prior to instillation of distilled water. The two groups were further divided and 15 minutes after simulated near drowning, the animals were either treated with surfactant (5 mL/kg instilled endotracheally) or not treated. Data were analyzed by repeated-measures analysis of variance. RESULTS: Near drowning had no significant effect on heart rate or blood pressure. Acidosis was not changed by surfactant; hypoxia was worsened by surfactant: 158 +/- 54 torr before and 66 +/- 19 torr after administration (p < 0.01). Compliance did not change as a result of surfactant administration. Only the animals (n = 3) in the normal O2 plus surfactant group died during the 90-minute post-near-drowning period of the study. CONCLUSIONS: Surfactant administration (5 mL/kg) in this animal model of freshwater near drowning offers no benefit over simple ventilation. Assessment of the merit of smaller volumes of surfactant should be investigated because administration of this solution may represent a second pulmonary insult, given current volume recommendations. PMID- 7497037 TI - When public health competes with individual needs. AB - A case of a homeless, alcoholic man with tuberculosis is presented. This case is used to illustrate the ethical dilemmas health care providers must face when the needs of society and the actions of the patient come into conflict. In addition to discussing historical cases of conflicts between individual rights and the needs of society, two models for solving the dilemma in the current case are considered. PMID- 7497039 TI - Ophthalmologic procedures in the emergency department--Part III: Slit lamp use and foreign bodies. AB - The emergency physician (EP) must be familiar with carrying out ophthalmologic procedures for evaluation and treatment of a multitude of eye complaints. This article is the last of three articles addressing ophthalmologic procedures of use by the EP. This article reviews the indications and the techniques for slit lamp examination of the eye and techniques of foreign body removal. Criteria for consultation also are addressed. PMID- 7497036 TI - Smoking cessation counseling by emergency physicians: opinions, knowledge, and training needs. AB - OBJECTIVES: To determine: 1) the extent of emergency physicians' (EPs') training in smoking cessation counseling; 2) their understanding of counseling and pharmacologic treatment techniques; 3) their current practices in screening, counseling, and referring patients who smoke; and 4) perceived barriers to routine smoking cessation counseling in emergency medical practice. METHODS: A 26 item questionnaire addressing the above issues was mailed to all 256 members of the Colorado Chapter of the American College of Emergency Physicians. RESULTS: Completed questionnaires were returned by 196 physicians (77% response rate). The majority of respondents were men (80%), practiced in urban settings (87%), and were board-certified in emergency medicine (82%). Most EPs lacked formal smoking cessation training (55%) and felt poorly prepared to counsel patients about smoking cessation (65%). A minority (27%) of the physicians reported routinely asking patients to quit smoking. The physicians with formal smoking cessation training were more likely to counsel and refer patients routinely (34% vs 20%, p = 0.03). The physicians cited the following barriers to routine smoking cessation counseling: a lack of time; a perception that patients are not interested; a belief that the ED setting is inappropriate for counseling; and a sense that counseling is ineffective. Lack of reimbursement was cited by only 13% of the respondents. The physicians who had formal smoking cessation training perceived fewer barriers to ED-based counseling. CONCLUSIONS: Emergency physicians have received little training in smoking cessation and perceive many barriers to ED based smoking cessation interventions. Not surprisingly, they infrequently take action to encourage or assist their patients to quit smoking. PMID- 7497038 TI - Bite to the left leg. Necrotic ulcer from a brown recluse spider bite. PMID- 7497040 TI - Chest discomfort, cocaine, and tobacco. PMID- 7497041 TI - Catheter-assisted intubation. PMID- 7497042 TI - Pediatric sedation and analgesia. PMID- 7497043 TI - Does your emergency medical services system need a device to confirm endotracheal intubation? PMID- 7497044 TI - Witnessed collapse and bystander cardiopulmonary resuscitation: what is really going on? PMID- 7497045 TI - Quo vadis: "scoop and run," "stay and treat," or "treat and street"? PMID- 7497046 TI - Impact of interventions for patients refusing emergency medical services transport. AB - OBJECTIVE: To evaluate the effect of a documentation checklist and on-line medical control contact on ambulance transport of out-of-hospital patients refusing medical assistance. METHODS: Consecutive patients served by four suburban ambulance services who initially refused emergency medical services (EMS) transport to the hospital were prospectively enrolled. In phase 1 (control phase), all patients who initially refused medical attention or transport had an identifying data card completed. In phase 2 (documentation phase), out-of hospital providers completed a similar data card that contained a checklist of high-risk criteria for a poor outcome if not transported. In phase 3 (intervention phase), a data card similar to that used in phase 2 was completed, and on-line medical control was contacted for all patients with high-risk criteria who refused transport. The primary endpoint was the percentage of patients transported to the hospital. RESULTS: A total of 361 patients were enrolled. Transport rate varied by phase: control, 17 of 144 (12%); documentation, 11 of 150 (7%); and intervention, 12 of 67 (18%) (chi-square, p = 0.023). Transport of high-risk patients improved with each intervention: control, two of 60 (3%); documentation, seven of 70 (10%); and intervention, 12 of 34 (35%) (chi-square, p = 0.00003). Transport of patients without high-risk criteria decreased with each intervention: control, 15 of 84 (18%); documentation, four of 80 (5%); and intervention, 0 of 33 (0%) (p = 0.0025). Of the 28 patients for whom medical control was contacted, 12 (43%) were transported to the hospital, and only three of these 12 patients (25%) were released from the ED. CONCLUSION: Contact with on-line medical control increased the likelihood of transport of high-risk patients who initially refused medical assistance. The appropriateness of the decreased transport rate of patients not meeting high-risk criteria needs further evaluation. PMID- 7497047 TI - Urban trauma transport of assaulted patients using nonmedical personnel. AB - OBJECTIVE: To describe one urban trauma transport system to clarify the impact of transport by nonmedical personnel on patient outcome. METHODS: Retrospective data were assembled over a six-year period through the use of the state trauma registry for an urban county served by seven state-accredited trauma centers. A subset of 4,767 consecutive assaulted patients was analyzed using the TRISS method to estimate survival probability. An unexpected death index (UDI), calculated as the difference between expected (TRISS method) and observed death rates, also was determined. Outcomes for patients transported by fire medics (FMs) vs nonmedical, police personnel (NPs) were compared. RESULTS: FMs transported 2,108 (44%) and NPs transported 1,356 (29%) of the injured assault victims. The FM-transported patients had a lower expected probability of survival than had the NP-transported patients (p < 0.001). This also was true within the penetrating-injury subgroup (p < 0.001), but not the blunt-injury subgroup. The observed death rate was higher for all the FM-transported patients than it was for the NP-transported patients (15% vs 11%; p < 0.01). The UDIs were not different overall, although the NP-transported patients who had blunt trauma had a significantly lower UDI (p < 0.01). CONCLUSIONS: NP transport of assaulted patients is generally associated with equivalent outcomes in comparison with FM transport in this urban environment. However, these data also provide evidence of an on-scene implicit triage with more severely injured patients generally transported by FMs. PMID- 7497048 TI - Bystander vs EMS first-responder CPR: initial rhythm and outcome in witnessed nonmonitored out-of-hospital cardiac arrest. AB - OBJECTIVES: To assess whether outcome and first-monitored rhythm for patients who sustain a witnessed, nonmonitored, out-of-hospital cardiac arrest are associated with on-scene CPR provider group. METHODS: A retrospective, cohort analysis was conducted in a suburban, heterogeneous EMS system. Patients studied were > or = 19 years of age, had had an arrest of presumed cardiac origin between July 1989 and January 1993, had gone into cardiac arrest prior to ALS arrival, and had received CPR on collapse. First-monitored rhythms and survival rates were compared for two patient groups who on collapse either: 1) had received CPR by nonprofessional bystanders (BCPR) or 2) had received CPR by on-scene EMS system first responders (FRCPR). RESULTS: Of 217 cardiac arrest victims, 153 (71%) had received BCPR and 64 (29%) had received FRCPR. The BCPR patients were slightly younger (62.4 vs 68.4 years, p = 0.01) and had slightly shorter ALS response intervals (6.4 vs 7.7 minutes, p = 0.02). There was no difference in BLS response time intervals or automatic external defibrillator (AED) use rates. The percentage of patients with a first-monitored rhythm of pulseless ventricular tachycardia/ventricular fibrillation (VT/VF) and the percentage of patients grouped by CPR provider who survived to hospital admission or to hospital discharge were: [see text] PMID- 7497049 TI - Colorimetric end-tidal CO2 detector for verification of endotracheal tube placement in out-of-hospital cardiac arrest. AB - OBJECTIVE: To evaluate the ability of a disposable, colorimetric end-tidal CO2 detector to verify proper endotracheal (ET) tube placement in out-of-hospital cardiac arrest, and to correlate semiquantitative CO2 measurements with the rate of return of spontaneous circulation (ROSC). METHODS: Prospective, observational study using a convenience sample of intubated out-of-hospital cardiac arrest patients. A disposable, colorimetric end-tidal CO2 detector was attached to the ET tube after intubation. In the absence of a colorimetric change, the paramedics reassessed the tube placement and could reintubate the patient. Tube placement was verified at the hospital. Paramedics were instructed to contact the base station and report the colorimetric change upon hospital arrival. ROSC was defined as restoration of a self-sustaining pulse until hospital arrival. RESULTS: Between December 1990 and May 1993, ET tubes were placed in 566 victims of out-of-hospital cardiac arrest. 541 of the 566 intubations (95.6%) were associated with a color change. In one case with a color change and out-of hospital clinical evidence of proper tube placement, the tube was determined to be in the esophagus at the hospital. Correct placement of the remaining 565 of 566 (99.8%) tubes was verified. Of the 566 patients who had a colorimetric change, 91 (16%) had ROSC vs one of 25 (4%) patients who did not have a color change. In one subgroup (n = 179), the degree of color change was highly associated with ROSC (p = 0.004). CONCLUSIONS: A disposable, colorimetric end tidal CO2 detector appears reliable in verifying proper ET tube placement in victims of out-of-hospital cardiac arrest. The degree of color change correlates with the probability of ROSC. PMID- 7497050 TI - Evaluation of a prototype esophageal detection device. AB - OBJECTIVE: To determine the ability of a prototype esophageal detection device (EDD) to identify esophageal misplacement of an endotracheal (ET) tube. METHODS: A prospective, randomized, blinded study of detection of esophageal intubation was conducted using 51 elective surgical patients who met inclusion criteria. A squeeze-bulb aspirator that creates a negative pressure of -80 to -90 torr was used for detection of esophageal intubation. The bulb should reinflate rapidly if the tube is in the noncollapsible trachea, but should not reinflate if the tube is in the collapsible esophagus. Each patient was prepared for surgery in the usual manner. The anesthesiologist placed an ET tube into the trachea. An identical tube was advanced the same distance into the esophagus. The tubes were labeled "A" or "B" according to a computer-generated random number list. An evaluator, who was blinded to the placement of the tubes, assessed one tube with the EDD. For most patients, a second evaluator, who was blinded to both tube placement and the results of the first evaluator, assessed the other tube. During evaluation, the tube cuffs were deflated. After data collection, the esophageal tube was removed and surgery was completed. RESULTS: All 45 esophageal tube placements were correctly identified. Thirty-five of the 40 ET tubes were correctly identified. Of the five ET tubes mislabeled, three were found in a mainstem bronchus. All had delayed bulb re-expansion. CONCLUSION: This prototype EDD is a useful method of identifying esophageal misplacement of an ET tube in anesthetized adult surgical patients. PMID- 7497052 TI - Family response to out-of-hospital death. AB - OBJECTIVE: To better understand the perceptions, needs, and responses of family members after an out-of-hospital death. METHODS: Over an 18-month period, phone interviews were conducted using a structured interview schedule modified from a similar study of survivors of in-hospital death. Subjects included 31 survivors of urban out-of-hospital deaths attended by paramedics from one ambulance company. Subjects were eligible if paramedics had arrived and death had been determined at the scene without transport. Survivors were interviewed 11 to 15 months after the death (mean = 12 months) to evaluate how well they coped with their loss, how they managed the experience of their loved ones' out-of-hospital deaths, and their feelings and beliefs about their loved ones' not being transported to a hospital. RESULTS: None of the survivors believed their loved ones should have been transported to a hospital, and only one believed that something more could have been done for the individual. Although many of the survivors had suspected their loved ones had died, 64% had been informed of the death by emergency medical technicians (EMTs) or firefighters. Most thought the EMTs had informed them in a professional (81%) and gentle/supportive manner (74%). Some (29%) still had unanswered questions about the death, but most (58%) were adjusting well and no one had a "poor" adjustment. CONCLUSION: In this small sample, survivors of out-of-hospital death were generally satisfied with the care their loved ones had received. None of the survivors believed their loved ones should have been transported to the hospital. They also believed the paramedics had been supportive and met their needs at the time of death. It appears that paramedics may be able to meet the needs of a patient's survivors by terminating out-of-hospital resuscitation efforts on the patient. PMID- 7497051 TI - Incidence, source, and nature of complaints received in a large, urban emergency medical services system. AB - OBJECTIVE: To document the incidence, source, and reasons for all complaints received by a large municipal emergency medical services (EMS) program. METHODS: A retrospective review of all complaints received during three consecutive years (1990-1992) in a centralized EMS system serving a large municipality (population 2 million). All cases were categorized by year, source, and nature of the complaint. RESULTS: In the three study years, EMS responded to 416,892 incidents with nearly a half-million patient contacts. Concurrently, 371 complaints were received (incidence of 1.12 per thousand); 132 in 1990, 129 in 1991, and 110 in 1992. Most complaints involved either: 1) allegations of "rude or unprofessional conduct" (34%), 2) "didn't take patient to the hospital" (19%), or 3) "problems with medical treatment" (13%). Only 1.6% (n = 6) were response-time complaints. Other complaints included "lost/damaged property," "taken to the wrong hospital," "inappropriate billing," and "poor driving habits." The most common sources were patient's families (39%) and the patients themselves (30%). Only 7.8% were from health care providers. CONCLUSION: Reviews of complaints provide information regarding EMS system performance and reveal targets for quality improvement. For the EMS system examined, this study suggests a future training focus on interpersonal skills and heightened sensitivities, not only toward patients, but also toward bystanders and family members. PMID- 7497053 TI - A case-based emergency medicine curriculum for senior medical students. AB - OBJECTIVE: To determine the effects of a case-based, core content-oriented emergency medicine (EM) curriculum on the basic EM knowledge of senior medical students. METHODS: All senior medical students rotating through the Milwaukee County EM elective during the 1992-1993 academic year were assigned specific chapter readings from a case-oriented EM textbook. A course curriculum consisting of goals and objectives for each chapter and two to three representative cases for the discussion topic also was distributed to each student. Interspersed with the cases was a series of questions directed at pathophysiology, diagnosis, management, and disposition. The EM faculty and residents conducted case discussions three times per week. All students completing the rotation were given a pretest at the beginning and a final examination at the end of the rotation. In addition, the students rated the textbook, coursebook, and lecture series at the end of the rotation using a five-point Likert scale. RESULTS: Seventy-five students rotated through the elective. The students showed a significant improvement in their EM knowledge base as judged by improvement in final examination scores compared with pretest scores (pretest score 62.2 +/- 7.1%; final examination score 76.2 +/- 6.3%; p < 0.0001). The mean change in scores was 14.8%, with a range of -1.6% to 34%. The students also rated the textbook, coursebook, and lecture series as effective, as shown by high median scores on a Likert scale. CONCLUSIONS: A case-based EM curriculum coupled with ED clinical experience improves basic EM diagnostic and management knowledge of senior medical students. PMID- 7497054 TI - Nonhemorrhagic pontine infarct in a child following mild head trauma. AB - A child who presented with hemiparesis secondary to a delayed non-hemorrhagic pontine infarction following mild head trauma is described. The results of the child's workup, including computed tomography (CT), were negative. The diagnosis of nonhemorrhagic pontine infarct was made by magnetic resonance imaging (MRI). The diagnostic evaluation excluded other possible etiologies of cerebral infarction, including vasculitides, CNS infection, congenital heart disease, hypercoagulable states, and demyelinating diseases. Although trauma cannot be proven as the cause of the infarct, other known causes of infarct were excluded. There are few cases of traumatic nonhemorrhagic cerebral infarction among children in the literature; none describes diagnostic MRI findings. MRI is important in these cases, because it may reveal delayed infarction from small vessel injury, which is not apparent on CT. This article discusses the etiology of and the diagnostic evaluation of pediatric cerebrovascular accidents and suggests the need for emergency physicians to consider trauma as a potential cause of delayed nonhemorrhagic cerebral infarct in children. PMID- 7497056 TI - The Australian Capital Territory Injury Surveillance and Prevention Project. AB - OBJECTIVE: To identify the representativeness and sensitivity of the data collected by the Australian Capital Territory Injury Surveillance and Prevention Project (ACTISPP) for ED injury cases seen in the Australian Capital Territory (ACT). METHODS: A retrospective descriptive study was conducted using a systematic sample of people injured in the ACT between January 20 and September 19, 1992, and who received their first treatment for this injury in an ACT ED. The ACTISPP surveillance system is dependent on ED patient or family completion and return of a comprehensive injury survey. The following population variables in the ACTISPP database were compared with those of the injured ED population, as determined by ED and hospital record review: age, gender, postal code of residence, nature of injury, body part injured, and whether the patient had been admitted to hospital. RESULTS: The sensitivity of the surveillance system for injuries seen in the ED was 31% (95% CI = 30-32%). The distribution of the ACTISPP sample did not differ from the expected distribution for sex or socioeconomic status. The distributions differed by age only in the 5-14-year age group (p < 0.001). With respect to the nature of injury and body part injured, the difference was significant (p < 0.001) but was no greater than 7% for any of the categories. Two percent of the surveillance sample were admitted to hospital, compared with an estimated 4% of the target population (p < 0.001). CONCLUSIONS: Approximately one-third of all incident injuries managed in the EDs of the ACT are included in the ACTISPP database. Although some sampling bias is introduced in the surveillance process, it does not appear to be of sufficient magnitude to compromise the project's goals. PMID- 7497055 TI - Pediatric vaginal bleeding. Urethral prolapse. PMID- 7497057 TI - The Health Consequences of Injury Questionnaire. AB - OBJECTIVE: To study the properties of a new survey instrument, the Health Consequences of Injury Questionnaire (HCIQ), for measuring the impact of injury on health and to compare the new instrument with the more detailed, previously developed Quality of Well-being (QWB) scale. METHODS: The HCIQ is a three-page health status questionnaire, suitable for self-administration. Similar to the QWB scale, the HCIQ measures health on a unitary scale where 1 represents optimum health and 0, death. The HCIQ was developed and studied in representative samples of injured adults from an Australian population. The HCIQ was administered concurrently with the QWB instrument to a sample of injured subjects, and the QWB scale scores were compared with estimated scores from the HCIQ using QWB scale weightings. The reliability of the HCIQ was tested on a further sample of injured subjects using a mailed test-retest design. RESULTS: Of the 211 subjects in the validity sample, 98 completed both the HCIQ and the QWB scales. The intraclass correlation coefficient for the scores obtained from the two questionnaires was 0.86 (95% CI = 0.66-1.00). The mean difference between the scores was 0.04 +/- 0.09 (SD). The HCIQ had a median completion time of 15 minutes. Of the 129 subjects in the reliability study, 53 completed the HCIQ on both postal administrations. The reliability coefficient was 0.90 (95% CI = 0.62-1.00). CONCLUSION: The HCIQ is a practical and reliable questionnaire for identifying the health consequences of injury that, when used with QWB weights, successfully predicts the QWB score. PMID- 7497058 TI - The Minor Injury Scale. AB - OBJECTIVE: To develop an injury scoring system suitable for non-life-threatening injuries. METHODS: A prospective cohort study was conducted using a representative sample of 1,396 injured adults to identify the outcomes of a range of injuries. Data were collected between March 12, 1992, and March 11, 1993, in the Australian Capital Territory, a geographically circumscribed urban community of 296,000 people served by two hospital EDs. Outcomes included "injury-related health status immediately following injury occurrence" and "accumulated health loss over the period of recovery." The injury-related health status of each subject at the time of the injury occurrence was measured using the Health Consequences of Injury Questionnaire (HCIQ), which records this outcome as a value on a unitary scale. Injuries were then stratified by injury type, as identified by the Abbreviated Injury Scale numerical identifiers. The median health index value for each type of injury was designated the injury severity score for that injury type. The "accumulated health loss over time until recovery" for each injury was calculated from repeated administrations of the HCIQ over a six-month follow-up period, and the median outcome value for each injury type was designated the injury morbidity score for that injury. RESULTS: Injury severity scores for 46 types of minor injury and injury morbidity scores for 39 types of minor injury were tabulated to form the Minor Injury Scale. CONCLUSION: This new injury scoring system provides a means of categorizing minor injury according to properties relevant to a public health approach to injury control. Expansion of the number of injury types scored requires further work, as does the testing of the results obtained to confirm the predictive power of the scales. PMID- 7497059 TI - Motorcycle helmet use: an annotated bibliography. PMID- 7497060 TI - Practice satisfaction, occupational stress, and attrition of emergency physicians. Wellness Task Force, Illinois College of Emergency Physicians. AB - OBJECTIVE: To define sources of job satisfaction and stress among emergency physicians and assess self-projected career longevity. DESIGN: A survey containing questions regarding emergency medicine (EM) practice satisfaction was mailed to 1,317 diplomates of the American Board of Emergency Medicine (ABEM). Specific sources of practice satisfaction and dissatisfaction, self-reported burnout or impairment, and plans for remaining in the specialty were assessed. Data were compared between two groups of physicians, namely, those residency trained in EM and those attaining certification through the practice or special category tracts. RESULTS: Of the physicians returning the survey, 25.2% stated that they felt burned out or impaired and 23.1% planned to leave the practice of EM within five years. Perceptions of burnout/impairment and plans to stop practice were associated with less overall practice satisfaction but were not significantly different between the two groups of physicians. Burnout/impairment was linked with psychiatric, drug, or alcohol problems and the feeling that EM had contributed to that problem. CONCLUSION: This study confirms the relatively high levels of projected attrition in EM and supports the perception that stress and burnout are associated with the specialty. Differences in job satisfaction and stress between those ABEM diplomates who were residency-trained in EM and those who became eligible for the board examination through practice or special category eligibility appear minor. PMID- 7497061 TI - Revascularization options: implications for critical transport. PMID- 7497062 TI - E-codes from emergency department records. PMID- 7497064 TI - Thrombocytopenic purpura. PMID- 7497065 TI - Hip fractures in the elderly: surveillance methods and injury control. AB - Injury is a major health problem in the United States that has been viewed primarily as a disease of the young. As a result, the devastating consequences of injury on the elderly population have not been sufficiently explored. Proximal femur fractures (hip fractures), common injuries in persons over the age of 65, carry a mild Injury Severity Score but are associated with high morbidity and mortality in the older population. The author provides a rationale based on clinical and research literature for increasing injury surveillance for hip fractures in the elderly in order that injury care and control can be improved. PMID- 7497066 TI - The president's message: trauma systems development. PMID- 7497067 TI - Developing an alias system for patient security. PMID- 7497069 TI - It's a New Year. How about some resolutions? PMID- 7497068 TI - The top ten topics for professional trauma outreach education programs. PMID- 7497070 TI - Diffuse axonal injury. AB - Diffuse axonal injury (DAI) is a severe form of traumatic brain injury that is seen almost exclusively following motor vehicle accidents. It is associated with immediate coma lasting from six hours to prolonged coma. The author reviews the mechanism of injury, significant neurological findings, neuropathology, and diagnostic criteria. Priorities for nursing care in the emergency department, critical care, and inpatient and rehabilitation units are outlined. PMID- 7497071 TI - [Effect of light intensity on binocular distance perception]. AB - Empirical investigations of perceived binocular distance often use an experimental setting in which real point-like light sources are presented in a completely dark surround. The results obtained, however, may be a consequence of the joint influence of binocular and monocular depth cues. Here, for the first time, the effect of (monocular) illumination cues on the locus of perceived equidistance is experimentally investigated by manipulating the luminous intensity of the stimuli. In a computer-controlled setup, small light-emitting diodes were presented in complete darkness. Five subjects judged the perceived distance of stimuli, which appeared in four directions and at five levels of luminous intensity, with respect to a fixed standard. Adaptive methods were used to determine perceived equidistance. Although the results show a significant effect of illumination cues, their influence on the locus of perceived equidistance turned out to be far less than previously supposed. For three subjects it was practically negligible. PMID- 7497072 TI - [Effect of task difficulty on hemispheric differences in visual image processing]. AB - Hemispheric specialization of visual imagery processes is a topic of recurrent debate. Recent theories have argued for a left postcentral center for the generation of visual images. Because of contradictory results the process of imaging has been divided into a left hemisphere component (visual imagery) and a right hemisphere component (supramodal spatial mental imagery). An alternative and more parsimonious account would be that the degree of difficulty of visual imagery tasks is responsible for hemispheric differences: easy tasks are solved faster within the left hemisphere because of a direct access to action control, whereas more difficult tasks rely on a right hemisphere specialization for spatial processing. In our experiment we varied the degree of difficulty by introducing concurrent tasks which were interspersed at random into the series of critical stimuli. Interspersing concurrent tasks prevents subjects from automatizing of critical processes and delays reaction times. For the critical visual imagery tasks we presented either upper or lower case letters to the different visual fields and asked for figural aspects of the corresponding lower or upper case letters, respectively. The interspersed stimuli concerned an estimation of the distance of the stimuli or the localization of the stimuli in the upper or lower part of the screen. We also presented upper and lower case letters in random order, thus forcing the subjects to shift between two different visual imagery tasks. In order to look for the effect of position-repetition priming upon visual field advantages we also analyzed the interaction of the position on the screen and its repetition with hemispheric advantages. In one of the visual imagery tasks, the experiments without concurrent tasks yielded a right visual field/left hemisphere advantage. On the other hand, interspersing other tasks resulted in a left visual field/right hemisphere advantage for visual imagery tasks, though none of the concurrent tasks showed a significant visual field advantage in itself. The same holds for combined and random presentation of the two different visual imagery tasks. The results therefore argue for a right hemisphere advantage for difficult visual imagery tasks, whereas easier tasks (monotonous presentation of only one visual imagery tasks) are processed faster in the left hemisphere. The random repetition of positions on the screen yielded a significant effect but had no influence on the visual field advantages found. PMID- 7497074 TI - [Priming effects in picture problems: preliminary solutions]. AB - Two experiments explored whether picture puzzles are an adequate instrument to investigate implicit memory for pictorial information. During the testing phase, the subjects had to identify hidden figures in picture puzzles. In a preceding learning phase, the priming conditions were varied systematically. In the first experiment, some subjects had to solve picture puzzles in the learning phase, whereas others made esthetic judgments (global processing) or estimated the number of triangles in the picture puzzles (local processing). In the second experiment, the subjects inspected copies of figures that were hidden at testing, modified versions of these figures, or their names in the learning phase. In the first experiment, the subjects of the different encoding conditions showed comparable priming effects. Picture puzzles that had already been processed or seen during learning were solved more often than new ones. Interview data revealed that subjects in the local or global processing conditions did not identify hidden figures at encoding. Furthermore, these subjects could not discriminate between old and new picture puzzles in a final explicit test of recognition. Thus, nonconscious storage of perceptual information that is not semantically interpreted may be sufficient to evoke priming effects. In the second experiment, the subjects in the different encoding conditions showed reliable priming effects, too. The presentation of the duplicates at encoding produced the greatest amount of priming. Effects of verbal priming, however, indicate that the solution of picture puzzles is not based solely on perceptual information. Depending on the priming conditions at learning, the solution of picture puzzles may be based primarily on data-driven processing or may be guided more heavily by conceptual information. It is concluded that perceptual, lexical, and/or conceptual information can contribute to the solution of picture puzzles. PMID- 7497075 TI - Collagen in liver fibrosis induced by ethanol. AB - Liver fibrosis is accompanied by significant increase of collagen content in this organ. Chronic intoxication with ethanol is probably the most popular cause of liver fibrosis. Some experimental and clinical data suggest that acetaldehyde (an intermediary product of ethanol oxidation) is responsible for stimulation of collagen biosynthesis in the liver. It increases collagen gene transcription in fibroblasts. About 90% of exogenous ethanol is oxidized in liver providing an enhanced amount of acetaldehyde which triggers the fibrotic response. PMID- 7497073 TI - [Interaction of differential anxiety states with analytic cognitive style]. AB - Interactions of anxiety states with modes of cognitive processing are investigated. It is assumed that the analytic thinking style of negative feeling states consists of probabilistic reasoning in the case of self-relevant anxiety states and of simple bipolar (yes-or-no) decisions in the case of physically threatening anxiety states. Sixty-six female student teachers (M = 23 years of age, SD = 2.7) served as subjects in a 3 x 2 x 4 mixed design. The subjects were divided into three groups of equal size. In each group a self-threatening or physically threatening anxiety state or a neutral feeling state was generated. Half of the subjects in each of the three groups were provided with either a probabilistic or yes-or-no answering format for a list of life events to induce a different style of cognitive processing. Repeated measurements of affect provided a pre-induction baseline, a manipulation check, and an assessment of affect change during task completion. The feeling states were assessed by means of scales for longitudinal studies of mood (Becker, 1988). The results show that the combination of self-threatening anxiety states with probabilistic judgments and- to a lesser degree--of physically threatening anxiety states with simple yes-or no judgments caused the maintenance of the respective feeling states. Neither the inverse combinations nor the processing styles alone led to similarly stable affective states. Anxiety states seem to be inherently connected with modes of cognitive processing that should be considered relevant for understanding affect task interactions and processes of mood repair. PMID- 7497077 TI - Ultrastructure of the hypothalamo-neurohypophysial system in rats exposed to lead. II. Neurohypophysis. AB - Ultrastructure of neurohypophysis was examined in rats allowed to drink for 6 weeks only solution of lead acetate. It was found the increased number of neurosecretory granules in axonal terminals, the signs of granulolysis in Herring bodies and the presence of axonal terminals with atypical, heterogeneous contents. The possible mechanisms of alterations observed in the whole hypothalamoneurohypophysial system affected by lead were discussed. PMID- 7497078 TI - Does hyperalimentation improve outcome in patients with severe head injury? AB - The purpose of this investigation was to evaluate the effectiveness of hyperalimentary therapy on the outcome in patients after severe head injuries. The observations were carried out in patients admitted within 24 hours after injury and divided into various alimentary groups (hyperalimentation, parenteral nutrition and enteral nutrition). The follow-up examination of patients' reactivity according to the Glasgow Coma Scale (GCS) performed 8-10 days after admission revealed improvement 1.2 +/- 0.9 pts in hyperalimentary subjects, while in the groups fed enterally or parenterally only 0.6 +/- 0.6 pts and 0.7 +/- 0.9 pts, respectively. In patients with severe head trauma and GCS score level lower than 5 the prognosis is unfavourable, irrespective of the treatment method. PMID- 7497076 TI - Ultrastructure of the hypothalamo-neurohypophysial system in rats exposed to lead. I. Magnocellular nuclei. AB - The effect of lead on ultrastructure of supraoptic and paraventricular neurons was studied in rats allowed to drink lead acetate solution instead of water during 6 weeks. Generally, the neurosecretory neurons appeared to be sensitive to lead dependently on its concentrations, but the ultrastructural patterns of the individual neurons varied from cell to cell and suggested the co-existence of the neurons in different functional states such as the increased secretory activity, adaptive responses or cellular degeneration. PMID- 7497080 TI - Vascularised patellar tendon graft in anterior cruciate ligament reconstruction. AB - Clinical results of 13 anterior cruciate ligament reconstructions by use of vascularised patellar tendon graft are presented. In all patients improvement was achieved resulting in good results in 62% cases (according to Lysholm score). Technical merits, fate of the graft and rehabilitation program are discussed. Described operation is recommended as the procedure of choice for the symptomatic anterior cruciate-deficient knee. PMID- 7497081 TI - Arthroscopic knee joint synovectomy in the treatment of early stages of rheumatoid arthritis. AB - Clinical results of 14 arthroscopic knee joint synovectomies are presented. They were done in grade II and III rheumatoid arthritis. Good and moderate results (according to Ogilvie-Harris) were obtained in 79% cases. Operative technique and postoperative management is presented. It was stated that arthroscopic synovectomy is most useful in II degree RA. PMID- 7497082 TI - Human histocompatibility complex (HLA). The distribution of HLA-ABC antigens and structure of exclusions in investigations on disputable paternity in population of north-eastern Poland. AB - In the present study 80 cases of disputable paternity analysed at the Department of Forensic Medicine in Bialystok in years 1990-1993 were reviewed. Gene frequencies of HLA antigens calculated for the north-eastern population of Poland are not significantly different from those reported for all-Polish population. Similarly, the most frequent HLA haplotypes calculated per 10,000 individuals for the studied population corresponded to those reported by others for central European population. In 13 cases (16.3%) paternity of a defendant was excluded. One case (7.7%) represented exclusions based on three antigens (of loci A, B and C), four cases (30.8%)-exclusions based on two antigens (of loci A and B) and eight cases (61.5%) exclusions based on one antigen (three of locus A and five of locus B). PMID- 7497079 TI - Ultrastructural evaluation of the rat parotid gland in the course of intoxication with lead acetate. AB - The purpose of this study was to evaluate the ultrastructural changes in the parotid gland of rats exposed to lead. The animals were given lead acetate in drinking water, at a concentration of 50 mg/dm3 Pb for periods of 2, 4 and 6 weeks. The ultrastructural examination showed that lead especially impaired mitochondria, caused accumulation of lipid droplets in the cytoplasm of acinar cells and inhibited the process of exocytosis of secretory granules. In the most extreme cases some acinar cells were destroyed. PMID- 7497083 TI - Cathepsin D--a main proteolytic enzyme of the bovine vitreous. AB - It was found that the bovine vitreous contains a proteolytic enzyme which demonstrates characteristic features of cathepsin D. It acts in acidic pH, actively digests denatured protein substrates and demonstrates a high susceptibility on the inhibitory action of pepstatin. It is localized in liquid part of the bovine vitreous. A role of this enzyme in physiology and pathology of the eye is discussed. PMID- 7497085 TI - Morphological heterogeneity of neurosecretory magnocellular nuclei of the guinea pig hypothalamus. II. Microscopic pattern and cytomorphological analysis of the paraventricular nucleus. AB - Position, shape and cytoarchitecture of the paraventricular nucleus were observed in 30 guinea pigs of both sexes, in serial frontal sections of the hypothalamus stained selectively for neurosecretion content, considering the sex of the animals examined and hypothalamus sides. Three morphological units of the nucleus were distinguished (parts: oral, principal and caudal) and four subunits (zones: medial, lateral, ventral and dorsal) as the basis for cytomorphological analysis. Morphology of PVN neurocytes was evaluated in 12 guinea pigs with regard to nine selected parameters. The results were formulated statistically. Cytomorphological comparisons were done between respective PVN units and subunits, depending on the side of sections in males. The results referring to the principal part of the right PVN in males and females were compared. Heterogeneity of the PVN-forming cellular populations was revealed. Cytomorphological differences were noted between PVN and SON. PMID- 7497084 TI - Morphological heterogeneity of neurosecretory magnocellular nuclei of the guinea pig hypothalamus. I. Microscopic pattern and cytomorphological analysis of the supraoptic nucleus. AB - Position, shape and cytoarchitecture of the supraoptic nucleus were observed in 30 guinea pigs of both sexes, in serial frontal sections of the hypothalamus stained selectively for neurosecretion content, with regard to the sex of the animals examined and hypothalamus sides. Three morphological units of the nucleus were distinguished (parts: oral, principal and caudal) and four subunits (zones: hypothalamic, optic, basal and central) as a starting point to work out a model of its cytomorphological heterogeneity. A detailed cytomorphological analysis was done within the distinguished morphological units and subunits of the supraoptic nucleus (SON) in 12 guinea pigs of both sexes. Using a light microscope equipped with a micrometric ocular, neurocyte morphology was evaluated on the basis of nine distinctive parameters. The results of the observation were formulated statistically. Cytomorphological comparisons were done between respective SON units and subunits, depending on the side of sections in males. The results referring to the principal part of the right SON in males and females were compared. Heterogeneity of the SON-forming-cellular populations was found. PMID- 7497086 TI - Natural radioactivity of building components and materials used in Bialystok province. AB - The paper presents the results of the measurements of natural radioactivity of building components and materials used in the North-East part of Poland performed by the Department of Biophysics, Medical Academy of Bialystok. The natural radioactivity (40K, 224Ra and 232Th) of 133 samples was measured and the qualifying coefficients f1 and f2 were calculated. All measured samples of different materials had radioactivity level below legally accepted value. The lowest values of f1 and f2 were observed in prefabricated building materials; and they were lower then in traditional red brick. Only one sample of furnace slag had increased value of natural radioactivity, which was higher than legally permitted level. PMID- 7497087 TI - Pulmonary permeability oedema. Methods of estimation of pulmonary capillary permeability. AB - Lung permeability oedema (acute respiratory distress syndrome-ARDS) is characterized by the increase of microvessels and epithelial permeability. In the estimation of pulmonary capillary permeability, the mathematical modeling is very useful. Some models and practical clinical methods concerning this problem are reviewed in this paper. PMID- 7497089 TI - The activity of granulocyte alpha-amylase in acute appendicitis. AB - The activity of alpha-amylase was measured in isolated granulocytes, serum and urine of 35 patients with acute appendicitis. The measurements were performed before operation and on the 7th day after operation. Slightly increased activity of alpha-amylase was found in the serum and urine of 15 patients. On the 7th day after operation the activity of this enzyme reached normal value. The activity of granulocyte alpha-amylase was elevated in 22 patients. In 2 of them the increased activity still maintained on the 7th day after operation. Positive correlation between the serum and granulocyte alpha-amylase activities was found. These observations allow to conclude that granulocytes are the source of increased alpha-amylase activity in the serum of patients with acute appendicitis. PMID- 7497088 TI - The effect of endotoxin on alcohol dehydrogenase (ADH) activity in the serum of ethanol intoxicated rats. AB - Serum alcohol dehydrogenase activity at different pH-values (8.8, 9.6 and 10.4) was investigated in induced endotoxemia (3 mg/kg) after chronic alcohol consumption in rats (60 days, 11.5 mg/kg, per day). It has been shown that ADH activity is higher after the single than after the triple injection of endotoxin given the healthy rats. Such a difference was not observed in the ethanol intoxicated rats. The single dose of endotoxin did not maintain the increased ADH activity after alcohol administration, however a triple injection of endotoxin maintained the activity but did not increase. This results show, that the highest ADH activity in all tested rats was observed at pH 9.6 and that chronic ethanol consumption did not cause an increase of the toxic effects of endotoxin on liver cells, measured by serum alcohol dehydrogenase activity. PMID- 7497090 TI - Some aquatic fungi of Hyphomycetes on tree leaves. AB - The presence of the aquatic Hyphomycetes in the rain water draining from the leaves of 17 species of trees (8 gymnospermous species and 9 angiospermous species) were studied. Evolution of 18 aquatic Hyphomycetes species was observed in the water draining from trees. The of most frequently encountered species of aquatic fungi Hyphomycetes in rain water draining from trees were Anguillospora pseudolongissima, Heliscus lugdunensis and Vargamyces aquaticus. PMID- 7497092 TI - A laboratory method for investigating influences on switching attention to task unrelated imagery and thought. AB - Thought-intrusions, automatic inferences, and other unintended thought are beginning to play an important role in the study of psychiatric disease as well as normal thought processes. We examine one method for the study of task unrelated imagery and thought (TUIT). TUIT likelihood was shown to be reliably measured over a wide range of vigilance tasks, to have high short-term and long term test-retest reliability, and to be sensitive to information processing demands. Likelihood of TUITs was shown to be different as a function of aging, hyperactivity, time of the day, and level of depression. Thus, we now can reliably measure the influence of endogenous and exogenous influences on TUITs. In addition, TUIT measurement was proposed as a minimally interfering and natural second task for determining resource utilization in a primary task. Finally, this method was offered as a reliable approach to quantification of such mental states as obsessions and drug craving and addiction. PMID- 7497091 TI - The lipid peroxidation products and the enzymes of antioxidant system in patients with diabetes mellitus. AB - The lipid peroxidation product level (conjugated dienes, malonyldialdehyde) as well as superoxide dismutase and glutathione peroxidase activity in diabetic patients were studied. It was found that diabetes mellitus is accompanied by an increase in serum conjugated diene and malonyldialdehyde level as well as by an increased malonyldialdehyde concentration in red blood cells. No changes in erythrocyte superoxide and glutathione peroxidase activity in diabetic patients were found. There were also no differences between diabetic patients with poor and good metabolic control in any of the examined parameters. PMID- 7497093 TI - Language, narrative, and autobiography. PMID- 7497095 TI - Repressed memories: the way we were? PMID- 7497094 TI - Metamemory and memory construction. AB - In this article, we present the contemporary conceptualization of metamemory as beliefs, accurate and naive, about memory. We discuss the implications of metamemory for memory construction in general and for suggestibility and the recovery of memories in particular. We argue that beliefs about memory influence (a) the probability that suggestions will be incorporated into memory and (b) judgments about the veracity of subsequent recollections. Implications for research on the role of beliefs in suggestibility and memory recovery are outlined. PMID- 7497096 TI - Empirical psychology and the repressed memory debate: current status and future directions. PMID- 7497097 TI - Telling the truth in the recovered memory debate. PMID- 7497098 TI - Trauma, dissociation, and clinical study as a responsible beginning. PMID- 7497099 TI - The recovered memory/false memory debate. Comment. PMID- 7497100 TI - Recovery memory/false memory polarities: balance and collaboration needed. PMID- 7497101 TI - Anosognosia in parietal lobe syndrome. AB - Patients with right parietal lesions often deny their paralysis (anosognosia), but do they have "tacit" knowledge of their paralysis? I devised three novel tests to explore this. First, the patients were given a choice between a bimanual task (e.g., tying shoe laces) vs a unimanual one (e.g., threading a bolt). They chose the former on 17 of 18 trials and, surprisingly, showed no frustration or learning despite repeated failed attempts. I conclude that they have no tacit knowledge of paralysis (or, if such knowledge exists, it is not available for this particular task). Second, I used a "virtual reality box" to convey the optical illusion to the patient that she was moving her paralyzed left hand up and down to the rhythm of a metronome, and yet she showed no sign of surprise. Third, I irrigated patient BM's left ear canal with cold water, a procedure that is known to shift that patient's spatial frame of reference by stimulating the vestibular system. Surprisingly, this allowed her "repressed" memory of the paralysis to come to the surface; she said she had been paralyzed continuously for several days. I suggest that the vestibular stimulation produces these remarkable effects by mimicking REM sleep. These patients also employ a whole arsenal of grossly exaggerated Freudian "defense mechanisms" to account for their paralysis. To explain this, I propose that in normal individuals the left hemisphere ordinarily deals with small, local anomalies by trying to impose consistency but, when the anomaly exceeds threshold, an interaction with the right hemisphere forces a "paradigm shift." A failure of this process, in patients with right hemisphere damage, might partially account for anosognosia. Finally, I present a new conceptual framework that may help link several psychological and neurological phenomena such as Freudian defense mechanisms, vestibular stimulation, anosognosia, memory repression, visual illusions, anterograde amnesia, REM sleep, dreaming, and humor. PMID- 7497102 TI - On the neurophysiology of consciousness: I. An overview. AB - How certain neural mechanisms (Mc) momentarily endow with subjective awareness (C) percepts and affects represented elsewhere is more likely to be clarified when structures essential to Mc are identified. The loss of C with bilateral thalamic lesions involving the intralaminar nuclei (ILN) contrasts with retention of C after large cortical ablations depriving C of specific contents. A role of ILN in the perception of primitive (i.e., not cortically computed) sensations is suggested by their afference of directly ascending pathways. A role for ILN in awareness of cortical activity is suggested by their widespread afference from cortex, a property shared with striatum. A role for ILN in volition is suggested by their heavy projection to striatum. Unlike striatum, ILN also project widely to almost all neocortex, enabling an effect on ideation; this last property is in common with other structures (e.g., locus coeruleus) but none of them has the same direct cortical afference. And passage through the reticular nucleus (nRt) of ILN efferents to cortex could impact the attention-selective action of nRt. It is suggested that the quickest route to a better understanding of C involves more intensive study of ILN. No other structure seems, in the light of our current knowledge, a more likely site for Mc. PMID- 7497106 TI - Child abuse, memory, and recall: a commentary. PMID- 7497104 TI - Steps toward healing: false memories and traumagenic amnesia may coexist in vulnerable populations. AB - Child abuse is surely the most agonizing psychological issue of our time. We decry the tendency to polarize around the either-or dichotomy of "recovered versus false memories," when both are likely to occur. Memory researchers seem to generalize from the mild, one-shot stressors of the laboratory to the severe repeated traumas reported by abused populations, an inferential leap that is scientifically dubious. Naturalistic studies show (a) some post-traumatic memory impairment (not just forgetting, but difficulty remembering in spite of repeated efforts); (b) dissociativity, such as emotional numbing, detachment, and the like; but also (c) increased suggestibility (Spiegel & Cardena, 1991). About 20% of the normal population is highly suggestible, and in these individuals it is trivially easy to show suggested amnesia, detachment, perceptual blocking, etc., as well as to suggest dramatically false memories. It is therefore vital to assess suggestibility and dissociativity in traumatized populations. Adult survivors of abuse may show both more false "memories" and more "false forgetting" than the normal population. PMID- 7497105 TI - Traumatic memories: do we need to invoke special mechanisms? PMID- 7497107 TI - The forgotten difference: ordinary memory versus traumatic memory. PMID- 7497103 TI - The trauma-memory argument. AB - The trauma-memory argument proposes that memories of childhood trauma can affect adult behavior outside awareness and that such unconscious memories can return to awareness even after long delays. Unfortunately, this conclusion is based on case reports of unknown representativeness and on clinical studies which are methodologically flawed or do not consider alternative explanations. Of particular concern is the general lack of independent verification of the ostensibly forgotten memories. The trauma-memory argument is plausible, in at least some respects, given what we know about the processes of remembering and forgetting, but considerably more research is needed before it can serve as a basis for scientifically sound clinical practice. PMID- 7497108 TI - The case of Dr. John D. Frame's first memory: historical truth and psychological distortion. PMID- 7497109 TI - Dreaming in the late morning: summation of REM and diurnal cortical activation. AB - Since the discovery that the characteristics of dreaming sleep are far stronger in Stage 1 rapid eye movement (REM) sleep than in any other biological state, investigators have attempted to determine the relative responsibility of the tonic versus the phasic properties of REM sleep for the different characteristics of dreaming--features such as the amount of information in the dream report, the brightness and clarity of the visual images, shifts in thematic continuity, and incongruities of image and meaning. The present experiment is designed to identify dream characteristics that are specifically associated with tonic changes in level of cortical activation within sleep. It samples reports of imagery and thought during spontaneous variations within one phase of the 24-h diurnal rhythm and across the REM-NREM (non-REM) sleep cycle in order to identify the independent and joint contributions of the two cycles to imagery and thought. The rising phase of the diurnal cycle in the late night and morning was estimated from clock time during the late night and early morning and was varied by delaying the sleep onset and waking time of the subjects. Considered together with other studies, the results suggest that the major determinant of vivid visual imagery and enhanced cognitive activity during sleep is a pattern of subcortical and cortical activation that is common to both the REM phase of the REM-NREM cycle and the activated phase of the 24-h diurnal wake-sleep cycle. PMID- 7497110 TI - A rediscovery of Sigmund Freud. AB - As part of his rediscovery of consciousness, Searle has recently provided an interpretation of Freud's account of consciousness, including the relation of consciousness to nonconscious mental occurrences [i.e., preconscious mental occurrences and unconscious (repressed) mental occurrences]. Regrettably, Searle's interpretation is based on a single paragraph from The Unconscious and serves to eliminate Freud's general view on these matters as being "incoherent." In the present article, I rediscover Freud's account and show that Searle has deeply misunderstood him, thus converting Freud into a "mental-eye" theorist of consciousness when Freud is actually an "intrinsic" theorist. I point out that Freud's and Searle's views on consciousness are actually similar, though Searle denies the existence of nonconscious mental occurrences. My discussion of Freud's account of consciousness addresses as well (a) Freud's understanding of emotions and feelings, including what he calls "misconstrued" emotions, and repression that works to suppress the development of affect, and (b) why Freud held that emotions and feelings never occur except consciously. I hope that my present contribution will be part of an intensive discussion that will proceed among psychologists of consciousness in a conscious and deliberate joint effort to develop the theory of consciousness. PMID- 7497111 TI - Belief in pain. AB - There is a traditional view of pain as a conscious phenomenon which satisfies the following two principles at least: Pain is essentially a belief- or cognition independent sensation, given for consciousness in an immediate way, and pain's unitary physical base is responsible for both its phenomenal or felt qualities and its functional, causal features. These are "The Raw Feels Principle" (P) and "The Unity of Pain Principle" (Q). Each is shown to be implausible. Evidence comes from recent pain research in a number of dimensions. A further argument against The Unity of Pain Principle is constructed by analogy with facts about blindsight. The consequences of rejecting P or Q are examined in light of (i) the traditional epistemological role of pain as exemplar of sensations thought to be foundational for knowledge, (ii) the prospects of a modularity theory of the pain system, (iii) recent accounts of pain among contemporary philosophers, (iv) a naturalist and realist conception of pain, and (v) views about the status of Folk Psychology. PMID- 7497112 TI - The sketchpad model. A theory of consciousness, perception, and imagery. AB - Subjective consciousness suggests a unity of the sensing and perceiving self that is difficult to reconcile with the multiplicity of sensory analyzers and the absence of a convergence zone in the brain. This has led--on the one hand--to the dead-end assumption of a unifying sentient homunculus and--on the other--to a denial of conscious unity. The sketchpad model presented here avoids this dilemma by viewing conscious thought as a selfreferent loop of neural activity, rather than as the information content of a fictitious set of output neurons. Use is made of the numerous neural pathways that originate at various cortical and subcortical areas and terminate in the thalamus. The model assumes that primary sensory inputs are modified by such feedback in the thalamic relay nuclei through hill-climbing processes that tend to optimize global responses. PMID- 7497114 TI - ORFs and genes: how strong a connection? AB - The length of an open reading frame (ORF) is one important piece of evidence often used in locating new genes, particularly in organisms where splicing is rare. However, there have been no systematic studies quantifying the degree of correlation between length of ORF, on the one hand, and likelihood of gene function, on the other. In this paper, techniques are derived to estimate the conditional probability of gene function, given ORF length, based on evidence both from the databases and from simulation. Several complete chromosomes of Saccharomyces cerevisiae have now been sequenced, and considerable effort is being expended on locating and characterizing the genes in these sequences. Thus, we illustrate the techniques for this organism. PMID- 7497115 TI - Algorithms for protein structural motif recognition. AB - The identification of protein sequences that fold into certain known three dimensional (3D) structures, or motifs, is evaluated through a probabilistic analysis of their one-dimensional (1D) sequences. We present a correlation method that runs in linear time and incorporates pairwise dependencies between amino acid residues at multiple distances to assess the conditional probability that a given residue is part of a given 3D structure. This method is generalized to multiple motifs, where a dynamic programming approach leads to an efficient algorithm that runs in linear time for practical problems. By this approach, we were able to distinguish (2-stranded) coiled-coil from non-coiled-coil domains and globins from nonglobins. When tested on the Brookhaven X-ray crystal structure database, the method does not produce any false-positive or false negative predictions of coiled coils. PMID- 7497113 TI - Computing with DNA. AB - We consider molecular models for computing and derive a DNA-based mechanism for solving intractable problems through massive parallelism. In principle, such methods might reduce the effort needed to solve otherwise difficult tasks, such as factoring large numbers, a computationally intensive task whose intractability forms the basis for much of modern cryptography. PMID- 7497116 TI - Four strikes against physical mapping of DNA. AB - Physical mapping is a central problem in molecular biology and the human genome project. The problem is to reconstruct the relative position of fragments of DNA along the genome from information on their pairwise overlaps. We show that four simplified models of the problem lead to NP-complete decision problems: Colored unit interval graph completion, the maximum interval (or unit interval) subgraph, the pathwidth of a bipartite graph, and the k-consecutive ones problem for k > or = 2. These models have been chosen to reflect various features typical in biological data, including false-negative and positive errors, small width of the map, and chimericism. PMID- 7497117 TI - Method for calculation of probability of matching a bounded regular expression in a random data string. AB - A method is presented for determining within strict bounds the probability of matching a regular expression with a match start point in a given section of a random data string. The method in general requires time and space exponential in the number of optional characters in the regular expression, but in practice was used to determine bounds for probabilities of matching all the ProSite patterns without difficulty. PMID- 7497118 TI - A note on scoring clones given a probe ordering. AB - We present an efficient algorithm for scoring clones given an ordering of probes under a schema proposed by Alizadeh et al. (1994) in the context of physical mapping with unique probes. The algorithm runs in time linear in the number of blocks of ones in the underlying sparse incidence matrix. A sparse and efficient algorithm for this task is important as it appears to be a central task in most algorithms for physical mapping. PMID- 7497120 TI - Modeling the polymerase chain reaction. AB - We introduce a mathematical model to treat the polymerase chain reaction (PCR), where we regard the accumulation of new molecules during a PCR cycle as a randomly bifurcating tree. This model enables us to compute an approximate formula for the distribution of the number of replications that have occurred between a pair of molecules, which depends on the efficiency lambda of the reaction, the number N0 of template molecules at the beginning of the PCR and the number c of PCR cycles. The reliability of the approximation is tested by computer simulations. Finally, to model the effect of the intrinsic error rate of the polymerase, we superimpose a substitution process on the tree. The resulting closed formula for the distribution of pairwise differences of sequences as a function of error rate mu and efficiency lambda can be used to estimate the error rate, if lambda is known. PMID- 7497119 TI - Classifying and counting linear phylogenetic invariants for the Jukes-Cantor model. AB - Linear invariants are useful tools for testing phylogenetic hypotheses from aligned DNA/RNA sequences, particularly when the sites evolve at different rates. Here we give a simple, graph theoretic classification for each phylogenetic tree T, of its associated vector space I(T) of linear invariants under the Jukes Cantor one-parameter model of nucleotide substitution. We also provide an easily described basis for I(T), and show that if I is a binary (fully resolved) phylogenetic tree with n sequences at its leaves then: dim[I(T)] = 4n-F2n-2 where Fn is the nth Fibonacci number. Our method applies a recently developed Hadamard matrix-based technique to describe elements of I(T) in terms of edge-disjoint packings of subtrees in T, and thereby complements earlier more algebraic treatments. PMID- 7497121 TI - The polymerase chain reaction and branching processes. AB - We construct a mathematical model for the polymerase chain reaction and its mutations using the theory of branching processes. Under this model we study the number of mutations in a randomly chosen sequence after n PCR cycles. A method for estimating the mutation is proposed and the variance of this estimator is studied. We also study the distribution of the Hamming distance between two randomly chosen sequences and a method for estimating the mutation rate based on pairwise differences is proposed. PMID- 7497124 TI - Combinatorial methods for DNA mapping and sequencing. PMID- 7497123 TI - Maximum discrimination hidden Markov models of sequence consensus. AB - We introduce a maximum discrimination method for building hidden Markov models (HMMs) of protein or nucleic acid primary sequence consensus. The method compensates for biased representation in sequence data sets, superseding the need for sequence weighting methods. Maximum discrimination HMMs are more sensitive for detecting distant sequence homologs than various other HMM methods or BLAST when tested on globin and protein kinase catalytic domain sequences. PMID- 7497125 TI - Physical mapping of chromosomes using unique probes. AB - The goal of physical mapping of the genome is to reconstruct a strand of DNA given a collection of overlapping fragments, or clones, from the strand. We present several algorithms to infer how the clones overlap, given data about each clone. We focus on data used to map human chromosomes 21 and Y, in which relatively short substrings, or probes, are extracted from the ends of clones. The substrings are long enough to be unique with high probability. The data we are given is an incidence matrix of clones and probes. In the absence of error, the correct placement can be found easily using a PQ-tree. The data are never free from error, however, and algorithms are differentiated by their performance in the presence of errors. We approach errors from two angles: by detecting and removing them, and by using algorithms that are robust in the presence of errors. We have also developed a strategy to recover noiseless data through an interactive process that detects anomalies in the data and retests questionable entries in the incidence matrix of clones and probes. We evaluate the effectiveness of our algorithms empirically, using simulated data as well as real data from human chromosome 21. PMID- 7497126 TI - Fragment collapsing and splitting while assembling high-resolution restriction maps. AB - In the process of constructing high-resolution restriction maps via greedy algorithms, a classical anomaly, known as fragment collapsing, introduces errors into the maps that impedes further map assembly. Fragment collapsing occurs when two different genomic fragments of approximately the same length and occurring in the digestion of two different overlapping clones are incorrectly identified as representative of a single genomic fragment. This introduces a single fragment of commensurate length into an incorrect position in the map. The present work describes techniques for detecting and correcting such anomalies. PMID- 7497122 TI - Prediction of function in DNA sequence analysis. AB - Recognition of function of newly sequenced DNA fragments is an important area of computational molecular biology. Here we present an extensive review of methods for prediction of functional sites, tRNA, and protein-coding genes and discuss possible further directions of research in this area. PMID- 7497127 TI - Ordering clone libraries in computational biology. AB - We consider a probabilistic model, due to Lander and Waterman and to Alizadeh, Karp, Newberg and Weisser, for the physical mapping of DNA molecules. Within this model, we answer precisely a question of Alizadeh et al. concerning the minimum number of probes required to reconstruct the entire ordering of a given clone library with high probability. We also examine the related problem of determining the least number of probes required to construct a "tiling" for the library. We give a fairly precise characterization for this number. PMID- 7497128 TI - Physical mapping by STS hybridization: algorithmic strategies and the challenge of software evaluation. AB - An important tool in the analysis of genomic sequences is the physical map. In this paper we examine the construction of physical maps from hybridization data between sequence tag sites (STS) probes and clones of genomic fragments. An algorithmic theory of the mapping process, a proposed performance evaluation procedure, and several new algorithmic strategies for mapping are given. A unifying theme for these developments is the idea of a "conservative extension." An algorithm, measure of algorithm quality, or description of physical map is a conservative extension if it is a generalization for data with errors of a corresponding concept in the error-free case. In our algorithmic theory we show that the nature of hybridization experiments imposes inherent limitations on the mapping information recorded in the experimental data. We prove that only certain types of mapping information can be reliably calculated by any algorithm. A test generator is then presented along with quantitative measures for determining how much of the possible information is being computed by a given algorithm. Weaknesses and strengths of these measures are discussed. Each of the new algorithms presented in this paper is based on combinatorial optimizations. Despite the fact that all the optimizations are NP-complete, we have developed algorithmic tools for the design of competitive approximation algorithms. We apply our performance evaluation program to our algorithms and obtain solid evidence that the algorithms are capable of retrieving high-level reliable mapping information. PMID- 7497130 TI - A new algorithm for DNA sequence assembly. AB - Since the advent of rapid DNA sequencing methods in 1976, scientists have had the problem of inferring DNA sequences from sequenced fragments. Shotgun sequencing is a well-established biological and computational method used in practice. Many conventional algorithms for shotgun sequencing are based on the notion of pairwise fragment overlap. While shotgun sequencing infers a DNA sequence given the sequences of overlapping fragments, a recent and complementary method, called sequencing by hybridization (SBH), infers a DNA sequence given the set of oligomers that represents all subwords of some fixed length, k. In this paper, we propose a new computer algorithm for DNA sequence assembly that combines in a novel way the techniques of both shotgun and SBH methods. Based on our preliminary investigations, the algorithm promises to be very fast and practical for DNA sequence assembly. PMID- 7497129 TI - Toward simplifying and accurately formulating fragment assembly. AB - The fragment assembly problem is that of reconstructing a DNA sequence from a collection of randomly sampled fragments. Traditionally, the objective of this problem has been to produce the shortest string that contains all the fragments as substrings, but in the case of repetitive target sequences this objective produces answers that are overcompressed. In this paper, the problem is reformulated as one of finding a maximum-likelihood reconstruction with respect to the two-sided Kolmogorov-Smirnov statistic, and it is argued that this is a better formulation of the problem. Next the fragment assembly problem is recast in graph-theoretic terms as one of finding a noncyclic subgraph with certain properties and the objectives of being shortest or maximally likely are also recast in this framework. Finally, a series of graph reduction transformations are given that dramatically reduce the size of the graph to be explored in practical instances of the problem. This reduction is very important as the underlying problems are NP-hard. In practice, the transformed problems are so small that simple branch-and-bound algorithms successfully solve them, thus permitting auxiliary experimental information to be taken into account in the form of overlap, orientation, and distance constraints. PMID- 7497131 TI - Short superstrings and the structure of overlapping strings. AB - Given a collection of strings S = [s1,...,sn] over an alphabet sigma, a superstring alpha of S is a string containing each si as a substring, that is, for each i, 1 < or = i < or = n, alpha contains a block of magnitude of si consecutive characters that match si exactly. The shortest superstring problem is the problem of finding a superstring alpha of minimum length. The shortest superstring problem has applications in both computational biology and data compression. The shortest superstring problem is NP-hard (Gallant et al., 1980); in fact, it was recently shown to be MAX SNP-hard (Blum et al., 1994). Given the importance of the applications, several heuristics and approximation algorithms have been proposed. Constant factor approximation algorithms have been given in Blum et al. (1994) (factor of 3), Teng and Yao (1993) (factor of 2 8/9), Czumaj et al. (1994) (factor of 2 5/6), and Kosaraju et al. (1994) (factor of 2 50/63). Informally, the key to any algorithm for the shortest superstring problem is to identify sets of strings with large amounts of similarity, or overlap. Although the previous algorithms and their analyses have grown increasingly sophisticated, they reveal remarkably little about the structure of strings with large amounts of overlap. In this sense, they are solving a more general problem than the one at hand. In this paper, we study the structure of strings with large amounts of overlap and use our understanding to give an algorithm that finds a superstring whose length is no more than 2 3/4 times that of the optimal superstring. Our algorithm runs in O(magnitude of S + n3) time, which matches that of previous algorithms. We prove several interesting properties about short periodic strings, allowing us to answer questions of the following form: Given a string with some periodic structure, characterize all the possible periodic strings that can have a large amount of overlap with the first string. PMID- 7497134 TI - Reconstruction of a string from substring precedence data. AB - The problem of reconstructing of a symbol string given the data about precedence of fixed length substrings arises in the method of nucleic acid sequencing by nested strand hybridization. We reformulate the problem in the graph-theoretical terms, describe the structure of the set of solutions, and present polynomial time algorithms that check existence and uniqueness of a solution or finiteness of the solution set, and then construct the solutions. PMID- 7497133 TI - DNA sequence recognition by hybridization to short oligomers. AB - A format 1 technology for performing massive hybridization experiments has been developed as part of the sequencing by hybridization (SBH) project. Arrays of tens of thousands of clones are interrogated with short oligomer probes to determine sets of oligomers that are present in individual clones. SBH requires highly discriminative hybridizations with a large number of probes. One of the main uses of a reconstructed DNA sequence is in a similarity search against databases of known DNA. We argue that sequence reconstruction, even partial, should not be performed for this particular purpose; we provide an information theoretic proof that the oligomer lists obtained from hybridization experiments should be used directly for similarity searches. We propose a similarity search method that takes full advantage of the subword structure of positively identified oligomers within a clone. The method tolerates error in hybridization experiments, requires fewer probes than necessary for sequencing, and is computationally efficient. To enable direct sequence recognition, we apply the recently developed method of sequence comparison that is based on minimal length encoding and algorithimic mutual information. The method has been tested on both real and simulated data and has led to a correct identification of clones based on hybridizations with 109 short oligomer probes. The method is applicable to hybridization data that comes from both format 1 and format 2 (sequencing chip) hybridization experiments. The sequence recognition method can provide targeting information for large-scale DNA sequencing by gel-based methods or by hybridization. PMID- 7497135 TI - Sequencing potential of nested strand hybridization. AB - The capability of the sequencing by nested strand hybridization (SNSH) method to sequence unseparated pools of DNA fragments was assessed in computer simulation experiments. The results demonstrate the high resolving power of the method and its tolerance to false-positive errors. We determine the optimal proportion between the fragment length and the pool size at a given length of oligonucleotide probes, compare SNSH to the standard SBH, and suggest the best experimental setting for the special case of sequencing of long isolated fragments. PMID- 7497132 TI - Reconstructing strings from substrings. AB - We consider an interactive approach to DNA sequencing by hybridization, where we are permitted to ask questions of the form "is s a substring of the unknown sequence S?", where s is a specific query string. We are not told where s occurs in S, nor how many times it occurs, just whether or not s a substring of S. Our goal is to determine the exact contents of S using as few queries as possible. Through interaction, far fewer queries are necessary than using conventional fixed sequencing by hybridization (SBH) sequencing chips. We provide tight bounds on the complexity of reconstructing unknown strings from substring queries. Our lower bound, which holds even for a stronger model that returns the number of occurrence of s as a substring of S, relies on interesting arguments based on de Bruijn sequences. We also demonstrate that subsequence queries are significantly more powerful than substring queries, matching the information theoretic lower bound. Finally, in certain applications, something may already be known about the unknown string, and hence it can be determined faster than an arbitrary string. We show that building an optimal decision tree is NP-complete, then give an approximation algorithm that gives trees within a constant multiplicative factor of optimal. PMID- 7497136 TI - The risk figure of 1/270. PMID- 7497137 TI - Effect of breast cancer screening after age 65. AB - OBJECTIVE: To assess the effect of mammographic screening for breast cancer in women aged 65 years or over. SETTING: The Swedish two county trial of screening for breast cancer, in which 77,080 women aged 40-74 (21,925 aged 65-74) were randomly allocated to receive regular mammographic screening for breast cancer, and 55,985 women aged 40-74 (15,344 aged 65-74) were allocated to an unscreened control group. METHODS: One group was screened every 33 months on average, except for those aged 40-49 at randomisation who were screened every 24 months. The control group was screened once at the conclusion of the trial. The main statistical analysis was the comparison of cumulative mortality with 13 years of follow up between the screened and control groups, in age groups 50-64 and 65-74, using Poisson regression. This was complemented by subsidiary analyses assessing the lead time, sensitivity, and predicted mortality from the size, node status, and grade of tumours diagnosed in the screened and control groups. RESULTS: In the age group 65-74 at randomisation there was a significant reduction in breast cancer mortality in the screened group, with a relative mortality of 0.68 and 95% confidence interval of 0.51 to 0.89. This was backed up by the results of the analyses of lead time, sensitivity, and tumour characteristics. CONCLUSIONS: Women aged 65 or more who are regularly screened can expect a reduced risk of dying from breast cancer. PMID- 7497138 TI - Revision of "NHS breast screening: the facts": an evaluation. PMID- 7497139 TI - Prenatal sonographic detection of isolated fetal choroid plexus cysts: should we screen for trisomy 18? AB - It has been established that the finding of a fetal choroid plexus cyst confers an increased risk of aneuploidy, predominantly trisomy 18. In view of this association many investigators have advocated amniocentesis when a choroid plexus cyst is identified by prenatal ultrasonography. A risk/benefit and economic analysis of such a policy is presented. When the following factors are taken into account-published data on the frequency of isolated choroid plexus cysts in second trimester fetuses, the risk of trisomy 18 in these fetuses, the population frequency of trisomy 18, the natural history and prognosis for these infants, and the risks and cost of amniocentesis--it is concluded that performing amniocentesis on second trimester fetuses with choroid plexus cysts is neither cost effective nor beneficial. For the detection of one infant with trisomy 18 that will survive past five months, 25 normal fetuses would be lost owing to amniocentesis. Identification of trisomy 18 by determining the presence of choroid plexus cysts in fetuses is not a worthwhile screening policy. PMID- 7497140 TI - Cost comparison of different methods of screening for cystic fibrosis. AB - OBJECTIVE: To compare the costs of several proposed methods of screening for cystic fibrosis. SETTING: England and Wales. METHODS: The costs of screening carried out at hospital antenatal clinics, general practitioner (GP) antenatal consultations, GP surgeries, and at work were estimated using data from demonstration projects. Couple screening, stepwise screening, and screening of individuals were considered. RESULTS: Couple screening at antenatal hospital clinics was the least expensive per carrier couple detected, amounting to 35,700 pounds (142,900 pounds for each potential cystic fibrosis fetus detected). The costs of the reagents (25 pounds per test) accounted for over 60% of this total. CONCLUSIONS: Antenatal screening, in addition to being the most cost effective method of screening, is also medically the screening method of choice as it provides information at the latest time when effective preventive action can be taken and at a time when all people to be screened are likely to be accessible. If the costs of the reagents could be reduced to 5 pounds (still higher than the costs of most diagnostic reagents) the cost for each pregnant carrier couple offered screening would be reduced by 50% to about 18,000 pounds, and the cost of offering screening to 684,000 pregnant couples in England and Wales would be about 9 1/2 m instead of 19 m pounds. PMID- 7497142 TI - Information leaflets. PMID- 7497143 TI - Construction of a community height surveillance programme: the Hackney growth initiative. AB - OBJECTIVES: To determine why, in the London Borough of Hackney before 1990, fewer children than expected were identified with remedial causes of short stature. To construct a practical model for height surveillance of 5 and 11 year old school entrants to improve the quality of child growth surveillance. SETTING: City and Hackney Borough, London, United Kingdom. METHODS: School nurses were trained by a clinical auxologist to measure children's height at school entry accurately and reproducibly. New procedures for measurement technique, plotting of data, referral, and audit were established. A reference manual was provided and a continuing training programme was started. RESULTS: During the first year the percentage of the target group measured was low. Changes in work practice led to improvements from 77% measured in the first year to 91% in the second year and 87% in the third year for 5 year olds. Improvements for 11 year olds were from 36% to 86% to 87% over the three years. Only 1.2% of 5 year olds and 2.6% of 11 year olds measured had height less than the third centile (compared with Tanner's height standards). CONCLUSIONS: School nurses measured height reliably. New audit procedures led to rapid changes in working practice and improvements in the percentage of children measured. The low numbers of short children previously identified with unrecognised abnormality may indicate an upward trend in height in this inner city population. PMID- 7497141 TI - Effect of parity on human chorionic gonadotrophin levels and Down's syndrome screening. AB - OBJECTIVE: To investigate further the association between parity and human chorionic gonadotrophin (hCG) levels during the second trimester and determine whether prenatal screening for Down's syndrome might be made more efficient by adjusting for this variable. METHODS: Measurements of hCG were analysed in relation to parity from a cohort of 16,675 singleton pregnancies with viable outcomes, and from 108 singleton pregnancies affected by Down's syndrome. Cigarette smoking during pregnancy and maternal age were also analysed in relation to hCG levels. RESULTS: For both unaffected pregnancies and pregnancies with Down's syndrome the median hCG levels were 7% lower in parous than in nulliparous women. A dose-response relation was also documented for the unaffected pregnancies. When para 0, para 1, and para 2 women were studied separately, hCG levels rose steadily as age advanced. Adjusting hCG levels for parity led to a 0.1% reduction in variance of the distribution of hCG measurements in unaffected pregnancies and to a negligible increase in Down's syndrome screening efficiency. CONCLUSION: The effect of parity is not sufficiently great to warrant routine adjustment of hCG levels as part of prenatal screening for Down's syndrome. PMID- 7497144 TI - How much paediatric HIV infection could be prevented by antenatal HIV testing? AB - OBJECTIVE: To estimate the reduction in the number of children infected with HIV that might be achieved by extending the provision of voluntary antenatal HIV testing. This effect would be mediated by increased numbers of women infected with HIV who receive an intervention to reduce the risk of vertical transmission (for example, zidovudine or caesarean section delivery), who use an alternative to breast feeding, or whose pregnancy is terminated. SETTING: London, United Kingdom. METHODS: Relevant data were derived from neonatal seroprevalence studies, obstetric and paediatric reporting schemes, and review of external information. Sensitivity analyses were performed for certain parameters. RESULTS: Of 106,000 births annually in London, an estimated 169 are to women infected with HIV whose infection is not currently recognised before pregnancy. An estimated 28 33 children born to these women will be infected. Precise prediction of the number of paediatric HIV infections that could be prevented is difficult because of uncertainty in certain factors, particularly the uptake of antenatal testing and the efficacy and acceptability of interventions to reduce prenatal or perinatal transmission. If a testing programme detected 70% of infected women, none of whom opted for a termination but all of whom exclusively bottle fed and received an intervention which halved the risk of transmission, about 12-16 (42 46%) paediatric HIV infections would be prevented annually. CONCLUSIONS: The estimated cost of preventing each paediatric infection is high, but this should be seen in the context of the lifetime health and social care costs for a child infected with HIV. The feasibility of selective testing should be considered when formulating policies on antenatal HIV testing. Programmes that are introduced should be audited to obtain better estimates of costs and benefits. PMID- 7497146 TI - Suggested screening guidelines for familial colorectal cancer. AB - Guidelines for screening for colorectal cancer in subjects with a positive family history of the disease, without the use of DNA based screening, are outlined. These suggestions are derived from (a) the experience of screening 644 subjects at an estimated lifetime risk of dying from colorectal cancer of 10% or more, in the St Mark's Family Cancer Clinic, over six years, and (b) a review of published studies, particularly incorporating the experience of the International Collaboration Group on Hereditary Non-polyposis Colorectal Cancer (HNPCC). Selection of subjects for surveillance depends upon the empirical evaluation of their risk of colorectal cancer, based upon family history details, with the exclusion of a diagnosis of familial adenomatous polyposis in the family. When DNA predictive tests for genes predisposing to HNPCC are available, surveillance can be directed at subjects with a germline mutation known to confer an increased genetic risk of colorectal cancer and discontinued in those at lower (or average) risk. In many subjects, however, DNA testing may still not be possible because of the failure to identify a predisposing mutation in an affected subject in their family. Any surveillance protocol can only be evaluated by long term follow up of those subjects in multiple centres. PMID- 7497145 TI - Effect of retesting with dietary restriction in Haemoccult screening for colorectal cancer. AB - OBJECTIVES: To determine the detection rate, false positive and false negative rates associated with a policy of retesting with dietary restriction after an initial positive Haemoccult screening test for colorectal cancer, in order to compare the cost effectiveness of such a policy with the alternative, in which all subjects with a positive test would proceed directly to diagnostic colonoscopy. METHODS: Over four years in a large randomised control trial in Nottingham 35,260 subjects had a mean of 1.5 screening rounds each at two-yearly intervals, and were followed up for a minimum of 27 months. During this period subjects with positive screening tests were asked to repeat the test with dietary restrictions. Estimates of costs of the initial screening and of diagnostic colonoscopy were used to estimate the cost for each cancer detected by the different policies. RESULTS: 1209 subjects had a positive initial screening test and 1033 (85.4%) completed the retests. Four hundred and ninety nine subjects were investigated and 89 cancers detected. In the 710 subjects with negative retests six interval cancers were diagnosed in the two years after screening. If these had been detected by screening under a policy of immediate colonoscopy, test sensitivity would have been improved from 53.6% to 57.2% (P = 0.02), but the cost for each cancer detected would have increased from pound 773 to pound 1509. CONCLUSION: Retesting with dietary restrictions reduces costs and maximises the benefit of limited colonoscopy resources, but results in a small but significant reduction in test sensitivity compared with a policy for immediate colonoscopy. PMID- 7497147 TI - Is psychological wellbeing impaired in long-term survivors of breast cancer? AB - OBJECTIVE: To measure anxiety and depression in long term survivors of breast cancer. DESIGN: Case-control study. SETTING: Follow up of the trial of early detection of breast cancer in the South West Surrey Health District. SUBJECTS: 331 patients with breast cancer aged 50-78, attending mammographic follow up, who had been invited to screening before diagnosis, and 584 controls who had attended the same clinic but did not have breast cancer. The sample included 290 pairs matched by screening attendance pattern and year of birth. MAIN OUTCOME MEASURE: Anxiety and depression scores on the Hospital Anxiety and Depression Scale. RESULTS: Significantly fewer symptoms of anxiety and depression were reported by patients with cancer than by controls and significantly fewer had high scores suggesting the presence of psychological morbidity meriting treatment. Among the patients with cancer anxiety was more common at the first annual follow up than at later visits. Symptom scores were not significantly related to the manner of detection or to the type of initial treatment. Potentially confounding social and personal factors did not account for the differences between patients with cancer and controls. CONCLUSION: The prevalence of anxiety and depression is not increased in long term survivors of breast cancer who are apparently free from disease and is not substantially affected by the manner in which a cancer is detected or by the treatment given. In the light of these findings it is difficult to justify large "quality adjustments" to the estimates of recurrence free years of life saved by screening. Those who counsel patients with breast cancer should be aware that although the initial distress can be severe, meriting psychological treatment, patients do recover their normal ability to enjoy life. PMID- 7497148 TI - The human element in medical screening. AB - An explanation is offered for the reasons why an experienced pathologist engaged in the screening of cervical smears should have issued a large number of false negative diagnoses. The explanation centres on the interactions which occur between successive judgments when a long sequence of similar stimuli are inspected. Briefly, those interactions have the character of an assimilation of each judgment to its predecessor, and the resultant process is capable of "running away". A procedure is suggested, in which the provision of immediate knowledge of results for a small proportion of the smears inspected would prevent such systematic misdiagnoses from recurring in the future. The implications of this natural experiment for medical screening in general are spelt out. PMID- 7497149 TI - Cervical smears: protection or prediction? PMID- 7497150 TI - Is screening for ovarian cancer worthwhile? PMID- 7497151 TI - Survival and interim end points in breast cancer. PMID- 7497153 TI - Setting standards for cervical screening fail-safe mechanisms: a target based on an audit of cytopathology laboratory records. AB - OBJECTIVE: To examine the efficacy of cytopathology laboratories in monitoring action following an abnormal cervical smear. SETTING: 11 screening laboratories serving 19 districts in the former North Western region of the United Kingdom. METHOD: 944 validated cases were identified at 11 laboratories. The screening history for each abnormal smear was obtained from the screening laboratory and compared with the records from the relevant family health services authority (FHSA) database. RESULTS: Laboratories held complete follow up records in 740 (78.4%) cases; the FHSA records extended this to 910 (96.4%) cases. Poor communication was the main problem; computerisation was not necessary to improve fail-safe procedures. CONCLUSIONS: Audit of fail-safe procedures is a valuable purchasing tool, identifying weaknesses and strengths, setting and maintaining standards. PMID- 7497152 TI - Opportunistic screening for abdominal aortic aneurysm. PMID- 7497154 TI - What are women told about Pap smears that lack endocervical cells? AB - BACKGROUND: Debate exists about the definition of what constitutes an adequate Pap smear and about the recommended rescreening interval for Pap smears lacking an endocervical component. This study aimed at determining whether women are currently informed about the endocervical status of their Pap smears and what rescreening recommendations are made to women whose smears lack endocervical cells. METHOD: Consecutive Pap smears lacking an endocervical component were identified from pathology records. After obtaining consent from the referring doctor, 165 women were interviewed by telephone. RESULTS: Only 110 (67%) of 165 women received active notification of their Pap test result and only six (4%) were aware that their smear lacked endocervical cells. Thirteen (8%) had been advised to have a repeat smear within three months. Nearly half the women reported that they would like more information about their result. CONCLUSIONS: It seems that current Pap smear notification patterns for women in New South Wales could be improved. One third are not actively informed at all about their results, and few are given detailed information about their Pap test results. Methods of enhancing the level of information women are given about their medical and screening tests need to be improved. PMID- 7497155 TI - Glaucoma screening. AB - National screening programmes probably cannot be justified at present given difficulties with test validity, manpower, and some doubts about the natural history of the disease. The opportunity cost could be prohibitive, especially when resources are badly needed in other areas of ophthalmology, such as cataract programmes, diabetic retinopathy, and low vision services. The situation could change with an improved test with greater validity for primary screening and such tests are currently under investigation. The results of these investigations will not be available for at least three years. Screening for eye disease at the primary care level begs questions about manpower which need to be answered not just for glaucoma screening but also for detection of diabetic retinopathy and visual impairment in the elderly. A new cadre of ophthalmic paramedics as is already in wide use in some developing countries, who amalgamate the skills of optometrist, orthoptist, and eye trained nurse, is an interesting possibility. Meanwhile, much has to be done to improve current case finding with agreed standards of examination and referral criteria and special facilities available for those at increased risk. PMID- 7497156 TI - The health service and personal costs of screening. PMID- 7497157 TI - Private costs associated with abdominal aortic aneurysm screening: the importance of private travel and time costs. AB - OBJECTIVES: To assess the importance of the private costs incurred by patients when making a judgment on the economics of screening for abdominal aortic aneurysm (AAA), and to explore the variation in such costs depending on screening location. SETTING: A district general hospital and general practitioner surgeries. METHODS: Four hundred and ninety nine consecutive subjects attending for AAA screening completed a questionnaire asking about travel arrangements for the journey to and from the clinic, the distance travelled, the time taken, the mode of transport, and any out-of-pocket expenses incurred. In addition, at the clinic each subject was asked what activities they had forgone in attending the clinic. Time was valued differently depending on whether work or leisure activities were forgone. The total private cost for each attender was calculated and comparison was made between attenders at hospital and at general practice. RESULTS: A significantly greater proportion of subjects were accompanied when attending hospital than when attending general practitioner (GP) surgeries. Most attenders travelled by car, but the journey time was significantly longer for those visiting hospital. The expected total private cost associated with attendance for AAA screening was 5.47 pounds. Attendance at GP surgeries had a lower private cost (4.21 pounds) than attendance at hospital (6.87 pounds). Only 7.3% of all men surveyed, and 6.5% of all companions, would have been taking part in some form of paid occupation if they had not attended for screening. CONCLUSION: Despite the fact that most attenders for AAA screening will be retired, the associated private costs are appreciable and should be considered in assessing the economics of such screening programmes. The level of private costs varied depending on the location of screening; clinics held at GP practices had lower private costs than those held at hospital. PMID- 7497159 TI - Prospects for the secondary prevention of colorectal cancer: screening by flexible sigmoidoscopy? AB - It may be useful to draw an analogy between the proposed screening programme for colorectal cancer and the cervical cancer screening programme. Both tumours show a spectrum of histological abnormalities consistent with a premalignant phase. The natural history of these premalignant lesions is poorly understood and although some will progress, if untreated, to invasive disease, most will not. Light microscopy cannot confidently distinguish which cases will progress and which will regress, and clinicians are therefore obliged to treat all. This will result in the destruction of many lesions of uncertain malignant potential. The secondary prevention of cervical cancer, although therapeutically efficacious, is inefficient. A lack of understanding of the natural history of intraepithelial neoplasia has frustrated attempts to develop rational referral criteria, and it is only now that the appropriate trials are being undertaken. The development of outpatient investigative and therapeutic procedures has resulted in many more women being referred for investigation and treatment, with predictable pressure on other services offered by gynaecologists, but no demonstrable saving of life. Similar uncertainties surround a screening programme for colorectal cancer. The principal concerns are not about the efficacy of polypectomy in interrupting the polyp cancer sequence, although uncertainties about the frequency with which cancer arises de novo do require that the effectiveness of this intervention is formally tested. Our major concerns are with compliance, and the management of the individual who tests positive--that is, who is found to have a distal polyp. Technological advances and operator enthusiasm may, as has happened with the cervical screening programme, lead to a relaxation in the indications for further investigation and treatment. Such a development would affect resources substantially if a population screening programme were in place. Nevertheless, there are grounds for believing that a screening programme for colorectal cancer, using sigmoidoscopy, might be successful in certain age groups if compliance was satisfactory. The scale of benefits may be comparable with those achieved by the breast screening programme. Our limited cost analysis, which relates to only to specific items of clinical activity, suggest that the mean cost for each case of cancer prevented will be about 8000 pounds sterling. These conclusions suggest that screening by flexible sigmoidoscopy merits serious consideration. It is also imperative, however, that consideration should be given to resolving some of the uncertainties about the clinical management and surveillance of those found to have distal polyps. PMID- 7497158 TI - Impact of prenatal screening and diagnosis on the epidemiology of structural congenital anomalies. AB - OBJECTIVE: To assess the impact of prenatal screening on the birth prevalence of three categories of structural congenital anomaly: abdominal wall defects (omphalocele and gastroschisis), renal agenesis/dysgenesis, and limb reduction defects. SETTING: Glasgow, Scotland, United Kingdom. METHODS: Data on the selected defects were obtained retrospectively from the population based Glasgow Register of Congenital Anomalies for the period 1980-91 inclusive. The register records all clinical or laboratory diagnoses of congenital anomaly in live births, stillbirths, and induced abortions occurring in women resident within the boundaries of the Greater Glasgow Health Board. The secular trends in the proportions of the defects diagnosed prenatally and terminated after screening, and in their prevalence at birth and during pregnancy, were examined. A total of 154,845 births were surveyed: 309 cases were identified in the selected anomaly categories. RESULTS: 83 cases of omphalocele/gastroschisis (5.4/10,000 births), 92 cases of renal agenesis/dysgenesis (5.9/10,000 births), and 134 cases of limb reduction defects (8.7/10,000 births) were found. Marked increases occurred over the study period in the proportions of cases diagnosed prenatally but not in the proportions terminated. The greatest difference between the prevalence at birth and during pregnancy was found for omphalocele. There were no significant secular trends in the prevalence of the selected defects. CONCLUSIONS: Prenatal screening has made a limited epidemiological impact on the prevalence of these defects. It has been moderately (but inconsistently) effective in the avoidance of births of infants with omphalocele/gastroschisis and renal agenesis/dysgenesis but not of limb reduction defects. Future efforts should be directed towards improving the technical aspects of the ultrasonographic detection of fetal abnormalities and exploring in detail, locally, the reasons for the varying pattern of decision making about termination of pregnancy among prospective parents. PMID- 7497162 TI - Sample size determination for a trial of breast cancer screening under age 50: population versus case mortality approach. AB - OBJECTIVE: To determine the efficacy of screening for breast cancer in women aged 40 to 49. METHODS: A large number of women will have to be enrolled to ensure adequate statistical power. Attention will be restricted to the deaths from breast cancer that occur among comparable groups of patients with breast cancer rather than to an analysis in which all women entering into the trial are considered at risk of dying from breast cancer (population mortality analysis). As the new trial is likely to be conducted in regions where screening has been established for all women from age 50 onward, comparability may be reached by including the breast cancers detected at the routine screening examination at age 50. CONCLUSIONS: When comparability of cases can be reached, such a case mortality analysis requires 15-30% fewer women to be enrolled than a population approach. PMID- 7497160 TI - Acceptance of screening for colorectal cancer by flexible sigmoidoscopy. AB - OBJECTIVE: To determine the acceptance rate of screening by flexible sigmoidoscopy in a sample of general practice patients aged 50 to 60, and the acceptability of the procedure among those undergoing the test. SETTING: Recruitment took place in selected general practices in Melbourne, Australia. Flexible sigmoidoscopies were performed at the local public hospital. PATIENTS AND METHODS: 187 patients aged between 50 and 60, and without bowel symptoms, were seen by one of 12 non-randomly selected general practitioners during the study period. At the conclusion of routine consultation general practitioners discussed colorectal screening with eligible patients and recommended that they attend a free screening clinic at the local hospital. The patients who attended the clinic underwent flexible sigmoidoscopy performed by experienced endoscopists. Clinic attenders completed a questionnaire which measured perceptions of the screening visit. RESULTS: The attendance rate was 49%. All clinic attenders were satisfied with the care received at clinic. Sixty five per cent of people experienced either no discomfort or mild discomfort, while 15% of people experienced moderate or severe pain. A bloated feeling was common (80%), with around 36% of people experiencing moderate or extreme bloating. Most people said that they were not at all embarrassed (64%), though 9% reported moderate or extreme embarrassment. Seventeen people (19%) were found to have polyps. CONCLUSIONS: Although the procedure was acceptable to those who underwent the screening test, further research on effective recruitment strategies is required if a full scale flexible sigmoidoscopy screening programme is to be considered feasible. PMID- 7497161 TI - A case-control study to evaluate efficacy of screening for faecal occult blood. AB - OBJECTIVES: Faecal occult blood testing is routinely used for early detection of colorectal cancer, but evidence of its efficacy in preventing death from colorectal cancer is limited. A case-control study was carried out to evaluate whether screening for faecal occult blood is associated with a reduced risk of fatal colorectal cancer. SETTING: A health maintenance organisation in western Washington State, which has offered its members faecal occult blood testing every two years since 1983. METHODS: Cases (n = 248) were members of the health maintenance organisation who died from colorectal cancer between 1986 and 1991. For each case, two control subjects, who did not die from colorectal cancer and who were similar to each case in age, gender, and year of enrollment at the health maintenance organisation, were randomly selected from the membership list of the year in which the case was diagnosed (n = 496). Information about episodes of faecal occult blood testing (including the location and reason for the test, and the evaluation of positive tests) and potential confounders was obtained from medical records. RESULTS: Cases were less likely than controls to have ever been screened (odds ratio (OR) = 0.7, 95% confidence interval (CI) 0.5 to 1.0), consistent with a beneficial impact of screening. There was little difference, however, for screening that had taken place within a three year period before diagnosis (OR = 0.9, 95% CI 0.6 to 1.2), the maximum interval during which most tumours ought to have been detectable by faecal occult blood testing. A reduction in risk was seen for home testing but not for office testing, and in individuals aged less than 75 but not in those aged 75 or older. Although most of the 21 controls with a positive faecal occult blood test underwent some additional testing, only five (24%) were evaluated with colonoscopy or air contrast barium enema. CONCLUSIONS: While there can be uncertainty as to whether specific faecal occult blood tests were performed as screening or diagnostic tests, those performed at home and in younger persons may be relatively less likely to be diagnostic tests that were misclassified as screening. Thus the modest reduced risk associated with faecal occult blood testing in these settings/persons may reflect genuine benefit. However, the presence of a reduced risk associated with a screening faecal occult blood test received in the past, well before a tumour or polyp might bleed enough to allow detection, is compatible with uncontrolled confounding. Interpretation is further complicated by the fact that a number of individuals in the study group who had positive test results underwent limited or no diagnostic testing. Thus our results should be interpreted with considerable caution. PMID- 7497163 TI - Screening for breast cancer in women aged under 50: mode of detection, incidence, fatality, and histology. AB - OBJECTIVE: To assess the effect of screening for breast cancer in women aged 40 49 in the Swedish two county trial, in terms of mortality reduction, advanced cancer reduction, mode of detection, and the histology of tumours detected. SETTING: The Swedish two county trial of screening for breast cancer, in which 77,080 women aged 40-74 (19,844 aged 40-49) were randomly allocated to receive regular invitation to mammographic screening for breast cancer, and 55,985 women aged 40-74 (15,604 aged 40-49) were allocated to an unscreened control group. METHODS: The screening interval in the younger age group was two years and in the older age group about three years. Statistical analysis of mortality and incidence rates was performed by Poisson regression. Relative survival was estimated using proportional hazards regression. RESULTS: The relative mortality for the group invited to screening compared with the control group was 0.87 (95% confidence interval 0.54 to 1.41) in the 40-49 age group, in close agreement with the relative incidence of advanced cancers. For Kopparberg county the relative mortality was 0.73 (95% CI 0.37 to 1.41) and for Ostergotland 1.02 (95% CI 0.52 to 1.99). The lesser effect in the 40-49 group as a whole was largely due to a higher rate of interval cancers in this age group, and the occurrence in Ostergotland of a higher number of cancers after randomisation, but before screening started, and in women who refused screening. The higher interval cancer rate was consistent with the higher proportion of ductal grade 3 and medullary cancers in women aged 40-49 at diagnosis. CONCLUSIONS: A major difficulty in screening women aged 40-49 is the rapid progression of a subset of tumours arising in this age group. Shortening the screening interval from two years would be necessary to achieve a higher mortality reduction. PMID- 7497164 TI - Independent double reading of screening mammograms. AB - OBJECTIVE: To evaluate the cost effectiveness of independent double reading of screening mammograms. SETTING: Prospective study of 18,817 women undergoing first or repeat screening in a population based programme in the Florence district. METHODS: Mammograms were independently double read by experienced radiologists. Subjects with mammographic abnormalities reported by at least one reader were recalled for diagnostic assessment. The mean increase in recall rate, cancer detection rate, and screening costs attributable to double reading was calculated. RESULTS: Eleven of 125 cancers were detected by only one reader. The mean increase in cancer detection rate attributable to double reading compared with single reading was 4.6% (95% confidence interval (CI) 1.1 to 8.9). From a total of 748 cases referred for diagnostic assessment, 196 subjects were referred by one reader only. The mean increase in referral rate attributable to double reading compared with single reading was 15.1% (CI 12.3 to 17.8). Double reading caused a marked increase in the cost for each woman screened -8.5% at the first screening and 6.2% at repeat screening and a more limited increase in the cost for each cancer detected -3.5% at the first screening and 2.7% at repeat screening. Cancers detected by only one screener were at an earlier stage than those detected by both screeners (P = 0.6, not significant). CONCLUSIONS: Independent double reading results in only a modest increase in the detection of cancers and therefore may not be cost effective. PMID- 7497165 TI - Renal medullary microcirculation: architecture and exchange. AB - The architecture of the renal medullary microcirculation is highly specialized. Consistent with their role in countercurrent exchange, the vessels (the vasa recta and the intervening capillaries) have high permeabilities to fluid and small hydrophilic solutes. The urea permeability of the continuous endothelium of the descending vasa recta (DVR) in the outer medulla is greatly enhanced by a urea transporter. Aquaporin channels have also been identified in these vessels. In spite of the absence of lymphatics from the inner medulla, fluid uptake from the interstitial fluid (ISF) through the fenestrated endothelium of the ascending vasa recta (AVR) appears to be driven by differences in hydrostatic and oncotic pressure. Because the AVR have high Lp's [10(-5) cm s-1 (cm H2O)-1] and are mechanically linked to surrounding structures, small increments of ISF pressures above the pressure within the AVR can drive significant volumes of fluid into AVR if ISF volume expands. The lower reflection coefficients to serum albumin of the AVR as compared with the DVR may be important in the clearance of interstitial plasma protein. Recent work on isolated DVR from the outer medulla has revealed that these vessels are capable of vasoconstriction and thus of regulating medullary blood flow. PMID- 7497166 TI - Gene-targeted mice in leukocyte adhesion research. AB - Physiologic and inflammatory leukocyte trafficking are controlled by adhesion molecules on leukocytes and endothelial cells. These adhesion molecules can be grouped into four major families: integrins, immunoglobulinlike molecules, selectins, and glycoproteins serving as selectin ligands. The recent advent of gene-targeting technology in embryonal stem cells has prompted the production of mutant mice that lack individual adhesion molecules or combinations thereof. Such gene-targeted mice permit studies into the roles of adhesion molecules in acute and chronic inflammation and of the regulation and interplay between different sets of adhesion receptors in vivo. Microcirculatory studies have been indispensable to our understanding of the importance of certain adhesion molecules for different steps of leukocyte recruitment in vivo. Targeting new genes and investigating mice with combined adhesion-molecule deficiencies will help to further clarify the molecular mechanisms of leukocyte trafficking in health and disease. This article reviews the insight gained through using adhesion molecule-deficient mice, emphasizing the role of microcirculatory studies in this research. PMID- 7497167 TI - Activation-dependent isolation and culture of murine pulmonary microvascular endothelium. AB - OBJECTIVE: Establish a reproducible method for the isolation and cultivation of murine pulmonary microvascular endothelium. To this end, we exploited the localized pattern of microvascular endothelial activation induced in vivo by inflammatory stimuli to isolate a subpopulation of endothelium for in vitro study. METHODS: Immunohistochemical analyses of the pulmonary vasculature of mice treated systemically with gram-negative bacterial endotoxin (LPS) demonstrated selective expression of VCAM-1 (CD106) in the endothelial lining of small collecting veins, venules, septal capillaries, and, infrequently, small arteries, which was not observed in control mice. Single cell suspensions prepared by enzymatic dissociation of peripheral lobular tissues dissected from the lungs of LPS-stimulated mice were incubated with a phycoerythrin-conjugated antimouse VCAM 1 monoclonal antibody (MK 1.91). Cells expressing this antigen were isolated by sterile fluorescence-activated cell sorting (FACS). Positive cell populations were collected and cultured for 1-2 weeks. When confluent, these primary cultures were further FACS enriched for endothelium, positively selecting for cells incorporating a fluorescent derivative of acetylated low density lipoprotein (Di I-Ac-LDL). RESULTS: The resulting population of cells (mouse lung endothelial cells, MLEC) were uniformly positive for the endothelial markers von Willebrand factor, thrombomodulin, and Dil-Ac-LDL uptake. MLEC readily formed tube-like structures when cultured on Matrigel and spontaneously demonstrated a sprouting phenotype on fibronectin or collagen matrices. MLEC retained responsiveness to cytokines (IL-1 alpha, IL-1 beta, TNF alpha, IFN gamma) up to at least eight passages from primary culture and demonstrated upregulation of E-selectin (CD62E) and P-selectin (CD62P) mRNA as early as 2 hr after LPS stimulation. Characteristic temporal expression patterns of cell surface E-selectin (maximal at 4 hr and declining toward baseline by 24 hr), VCAM-1 (maximal at 6-8 hr and remaining elevated for 24-48 hr), and ICAM-1 (maximal at 6-8 hr and maintained at 24 hr) were observed when cultured MLEC were treated with recombinant murine TNF alpha or recombinant human (rh) IL-1 alpha or rhIL-1 beta. The rolling, adhesion, and transmigration of human polymorphonuclear leukocytes was markedly increased on cytokine-activated MLEC monolayers under defined flow conditions. CONCLUSION: The strategy of activation-dependent isolation allows for the reproducible selection of a specific subset of microvascular endothelial cells for in vitro study. This experimental approach should further facilitate study of the functional heterogeneity of endothelium and its pathophysiologic dysfunction. PMID- 7497169 TI - Pulmonary microvascular pressure profile during development of hydrostatic edema. AB - OBJECTIVE: To measure, in intact closed chest, the pressure in the pulmonary microvasculature during transition to mild interstitial edema. METHODS: In anesthetized spontaneously breathing rabbits, the pulmonary artery and left atrium were cannulated. Pleural windows were prepared to view the superficial pulmonary microvascular network through the intact parietal pleura. After intravenous infusion of 96.4 +/- 12.3 ml of saline at a rate of 0.5 ml/kg h, the hydraulic pressure in the pulmonary microvessels (15-240 microns in diameter) were measured using glass pipettes driven through the pleural window and connected to a servonull system. RESULTS: After saline, plasma protein concentration decreased from 6 +/- 1 to 4.8 +/- 0.5 g/dl; pulmonary arterial and left atrial pressures averaged 22.3 +/- 6.4 and 2.3 +/- 2 cm H2O in control and 23.1 +/- 4.2 and 4.2 +/- 2 cm H2O after infusion. After saline loading, 16.4% of total pressure drop occurred from pulmonary artery to 80-microns arterioles, 60.3% in 30-80 microns arterioles, 6.9% from 30-microns arterioles to 30-microns venules and 16.4% in the downstream segment. CONCLUSIONS: Mild interstitial edema induced, with respect to control, constriction of small arterioles and capillary recruitment to maintain a low capillary pressure. Hence, in initial edema, pulmonary circulation prevents further fluid filtration, acting like an intrinsic safety factor to delay development of severe edema. PMID- 7497168 TI - Thromboxane contributes to submaximal coronary dilation during myocardial ischemia. AB - OBJECTIVES: Thromboxane has been shown to contribute to coronary constriction in conduit coronary arteries during platelet aggregation at the site of a critical stenosis. Previous studies from our laboratory suggest that following a critical coronary stenosis, persistent vasomotor tone occurs. We tested the hypothesis that thromboxane is responsible for that increased tone. METHODS: To test this hypothesis, 14 mongrel dogs of either sex were anesthetized and subjected to a critical coronary stenosis where distal coronary perfusion pressure was reduced to 36 +/- 2 mm Hg. During the coronary stenosis, SQ 29,548 (thromboxane/endoperoxide receptor antagonist) was administered intravenously (0.2 mg/kg and 2.0 mg/kg). Coronary microvascular responses were observed by directly visualizing the epicardial microcirculation. Diameters were measured using intravital microscopy coupled to stroboscopic epi-illumination and jet ventilation to compensate for cardiac and respiratory-induced motion. RESULTS: Coronary microvessels were divided into small (< 150 microns) and large (> 150 microns) arterioles. During SQ 29,548 administration, small coronary arterioles demonstrated no additional dilation during a critical coronary stenosis. In contrast, coronary microvessels > 150 microns demonstrated a dose-dependent vasodilation to SQ 29,548 (0.2 mg/kg: 6 +/- 2%; 2.0 mg/kg: 11 +/- 4%; P < 0.05 vs. no change). A time control study in six additional animals demonstrated no significant microvascular diameter changes following a critical stenosis over the time course of SQ 29,548 administration. CONCLUSION: Endoperoxides contribute to poststenotic microvascular vasoconstriction in vessels 150-300 microns. PMID- 7497170 TI - Heterogeneity of red blood cell velocity in skeletal muscle decreases with increased flow. AB - OBJECTIVE: Effective material exchange between blood and tissue depends on the heterogeneity of microvascular flow. The objective was to address inconsistencies between intravital studies regarding this dependency. We tested the hypothesis that heterogeneity of red blood cell velocity (VRBC) in capillary beds varies with the strength of metabolic stimulus and with capillary bed geometry. METHODS: We used videomicroscopy to measure VRBC in a bed of 10-24 capillaries at the surface of extensor digitorum longus (EDL) muscle in anesthetized rats. The coefficient of variation (CV = standard deviation/mean; an index of spatial heterogeneity) was computed in the same bed before and after (i) 1, 2, 4, or 8 Hz supramaximal muscle contraction or (ii) adenosine superfusion (10(-7)-10(-3) M). Beds with or without arteriolar-venular capillary shunts were used. RESULTS: Although control VRBC differed between beds (shunt: 232 microns/s; no shunt: 130 microns/s), the percentage increases in postcontraction VRBC did not (range: 111 326%). In both beds, control CV varied greatly (overall range: 28-117%) and 2-8 Hz muscle contractions reduced CV significantly by 25%. Similar results were obtained for adenosine. In confirmatory experiments using the rat cremaster muscle, contractions (4 Hz) and adenosine (10(-4) M) also reduced CV. Based on all data, CV = 63-0.022 VRBC (r = 0.82, P < 0.001). CONCLUSIONS: The heterogeneity of VRBC decreased with metabolic stress, regardless of capillary bed geometry. We propose that both the large variability in control CV and the relatively shallow dependence of CV on velocity could be responsible for the present inconsistencies between intravital studies. PMID- 7497171 TI - The H2O2-generating enzyme, xanthine oxidase, decreases luminal Ca2+ content of the IP3-sensitive Ca2+ store in vascular endothelial cells. AB - OBJECTIVE: Xanthine oxidase inhibits agonist-stimulated Ca2+ signaling in calf pulmonary artery endothelial cells by an H2O2-dependent mechanism. We investigated the effect of xanthine oxidase on luminal Ca2+ content of the inositol-1,4,5-trisphosphate (IP3)-sensitive Ca2+ store. METHODS: Luminal Ca2+ content was estimated from the net release of Ca2+ activated by 2,5-di-t butylhydroquinone (BHQ), an inhibitor of microsomal Ca2+ pumps. RESULTS: Initially, xanthine oxidase depleted the IP3-sensitive Ca2+ store of releasable Ca2+, but with more prolonged incubation, the enzyme also depleted non-IP3 sensitive stores. In addition, xanthine oxidase inhibited capacitative Ca2+ influx. Similar results were observed when thapsigargin was substituted for BHQ. CONCLUSIONS: Depletion of luminal Ca2+ content within the IP3-sensitive Ca2+ store contributes to xanthine oxidase inhibition of Ca2+ signaling in vascular endothelial cells. PMID- 7497172 TI - [Sleep disorders and Parkinson disease: study of a series]. AB - Disturbances of sleep are frequent and varied in patients with Parkinson's disease. In an attempt to find out about these disturbances, we carried out interviews using the Vanna and Mazzuia questionnaire, modified for forty-five sufferers of Parkinson's disease, and forty healthy adults. We researched sleep habits and disturbances, as well as the consumption of hypnotics. We found that both groups had similar habits, although those with PD altered according to the stage of development of the disease, thus difficulty in staying sleep and parasomnia were more frequent in PD sufferers and worsened in the most advanced cases. On the other hand the quality of night sleep and the consumption of hypnotics interfered in the daytime efficiency of those suffering from Parkinson's Disease. PMID- 7497176 TI - [Etiological and prognostic factors in the late onset epilepsy]. AB - The etiological and prognostic factors of epilepsy starting in adulthood have been studied globally and especially in the case of that group of patients whose epilepsy begins after the age of 60. However, a specific study of patients whose epilepsy has begun after the age of 40 has not been made so far. In this report the above-mentioned factors are analysed in patients who were examined consecutively in our center between May 1986 and December 1990, all of whom had had their first epileptic attack after the age of 40. During this period 85 patients [55 (74.07%) women: 30 (35.3%) men: average age: 65.41 +/- 13.23] fit in to this category. The starting age was 61.6 +/- 13.5. The most frequent causes of the epileptic attacks were cerebrovascular accident (41%) and alcoholism (16%). In 1% of cases, the cause of the attacks could not be identified. Antecedents such as chronic alcoholism and tobacco addiction, were more frequent among the men (30.9% against 3.3% p: 0.014 and 49.1% against 0% p: 0.0000, respectively). The most frequent etiology in the age group comprised of patients between 40 and 50 years of age was alcohol and drug consumption. In the group of patients aged between 70 and 80, the most frequent finding in computerised cranial tomography was encephalic infarction. The factor linked to an unsatisfactory development of the epileptic process was nonobservance of therapeutical recommendations registered clinically as well as by means of the calculation of plasmatic levels of antiepileptic medicaments (p: 0.0000). Consumption of alcohol was the factor linked in a significant manner to the nonobservance of therapeutical recommendations (p: 0.0128). PMID- 7497173 TI - Intracavernous pharmacotherapy for management of erectile dysfunction in multiple sclerosis patients. AB - Intracavernous self-injection of vasoactive drugs is a well accepted therapy for the management of erectile dysfunction in neurogenic disorders. We present the results of a self-injection program in seven MS patients who were admitted in Institut Guttmann of Barcelona and who presented with an insufficient erection. All patients showed an excellent erectile response and had penile rigidity sufficient for sexual intercourse during acceptable time, with minimal complications. A patient reported improvement of erectile capacity so as not to need another cavernously injection for 2-3 months. PMID- 7497175 TI - [Paramedian bithalamic infarct syndrome: report of five new cases]. AB - Five patients were examined suffering from bilateral paramedian thalamic infarction, caused by occlusion of the posterior paramedian thalamo-subthalamic arteries, when they begin from one single pedicle. All cases began with obnubilation or transitory coma, followed by hypersomnia. Four patients showed vertical gaze paralysis, and the fifth vertical nystagmus. In three cases, nuclear lesion of the III cranial nerve was observed along with alteration of the photomotor reflexes, and there was miosis in one case. All were suffering from weakness in one or another limb or facial paresis and generalised acute hypotonia: only one patient had hemihypostesia. All five had dysarthria, ataxy and dysmetria, one had asterixis and two spasmodic crying. Between 5 and 12 months later, one had akinetic mutism and vertical gaze paralysis as the most noteworthy signs. The neuroradiological images show a bilateral ischemic lesion in the paramedian thalamic region, which extends in some cases to the anterior nucleus and in one case to the pulvinars; the lesion continues through the subthalamic regions and the medial part of the mesencephalic tegmentum, with a clear extension to the medial region of the cerebral peduncles in three cases and to the tectum in one case. PMID- 7497174 TI - [Latency in the hospital admittance of stroke]. AB - OBJECTIVE: the hospital referral of 200 patients with cerebrovascular disorders in order to evaluate the possibility of treating acute cerebral infarction. METHOD: by means of a personalized interview an evaluation was made of the time that had passed between the start of the symptomatology and the time the patient went into hospital. The results were then grouped into intervals and compared to the different nosological entities of cerebral vascular pathology. The data were analysed using non-parametrical statistical tests (chi 2). RESULTS: of the 200 patients examined, 79.5% had cerebral ischemy (15.1% AIT, 84.9% infarction) and 20.5% intercranial haemorrhage (92.7% intercerebral haemorrhage). Of the patients suffering from cerebral infarction, 27.4% involved lacunary infarctions, 22.2% cardio-embolic infarctions, and 50.4% atherothrombotic infarctions. In 57% of cases the pathology was located in the carotid region and in 35.5%, in the vertebrobasilar region. 46.5% of the patients suffering from cerebrovascular disorders went into hospital less than 3 hours since the start of the symptomatology. 60.7% of the patients with cerebral infarction arrived within 6 hours. When the latency between the different nosological entities was compared, significant statistical differences were only detected between cardio-embolic infarctions and the other infarctions (p = 0.04) and between carotid and vertebrobasilar pathology (p = 0.01). CONCLUSIONS: based on the hospital referral time, the design of new clinics in our field for the treatment of patients suffering acute cerebral infarction is viable, above all as far as those of cardio-embolic origin and located in the carotid region are concerned. PMID- 7497178 TI - [A study of mortality in patients with acute cerebrovascular pathology in the Castellon General Hospital]. AB - OBJECTIVE: To analyse the hospital death rate caused by acute cardiovascular disorder and their factors conducted. METHODS: We made a retrospective review of all patients suffering from this pathology who had gone into the Casualty Ward of the Castellon General Hospital in the course of 1991: we also looked for all patients with acute cardiovascular disorder who had died in the hospital. We studied the risk factors for the patients referred using a univariant analysis, comparing those still living with those who had died. RESULTS: We find 514 patients diagnosed as having acute cardiovascular disorder, with an stroke/TIA incidence rate of 188 cases per 100,000 inhabitants. The proportion of referred patients was 52%. Among these, the death rate was 27.8%. Factors affecting the death rate were the age of the patients, the reduction in the level of consciousness, and lateral gaze at the start of the stroke. CONCLUSIONS: We believe that the high death rate in our selection was due to the severity of the strokes and the advanced age of the patients, as was in accordance with the methodology used in the gathering of data, which made it possible to include patients who died early. PMID- 7497177 TI - [Treatment of the partial epilepsy in the first two years of life. Experience in 26 cases]. AB - We performed a retrospective study with 26 patients with partial epilepsy (16 male, 10 female), aged between 1 and 24 months. We excluded those patients with seizures secondary to cerebral and extracerebral acute diseases, febrile convulsions, infantile spasms and other epileptic syndromes frequently diagnosed at this age. Etiology was of prenatal origin in 8 cases, perinatal in 9, postnatal in 3, and 6 were cryptogenic. They were all initially treated with phenobarbital (PB), 5 mg/kg/day. If seizures did not disappear, clonazepam (CZP) 0.1 mg/kg/day was added. Fifteen patients were controlled with PB in monotherapy, 7 need the association of CZP, and 4 patients could not be controlled. Among the last ones, 2 were controlled with PB and phenytoin, and the other 2 showed no response to any other drugs. Side effects (drowsiness and increased oropharyngeal secretions) were mild and transient. According to our experience, PB, alone or associate with CZP, is a safe and efficient treatment for seizures in symptomatic and cryptogenic partial epilepsy. PMID- 7497179 TI - [Early infantile epileptic encephalopathy]. AB - Early infantile epileptic encephalopathy (EIEE) with burst-suppression (Ohtaharas syndrome) is a rare type of epileptic encephalopathy in infancy and represents the earliest type of age-related symptomatic generalised epilepsy. We present 4 cases of EIEE fulfilling the classic diagnostic criteria, excluding the cases with segmentary and erratic myoclonic, characteristic features of Early Myoclonic Encephalopathy. All the patients were females. Pregnancy and delivery were normal and no other cases were observed in the families. One of the patients had a normal twin brother. Seizures began in all the cases before the 10th day of life. The initial EEG showed burst-suppression pattern, lasting for an average of 2.3 months of age (1.5-4 months). Seizures were initially resistant to therapy, although 2 cases showed partial response. None of the patients evolved to West syndrome. A case died at 5 months of age. No etiologic factors were found in two cases, but hemimegalencephaly was detected in the other two patients; in one of them seizures disappeared after corticectomy. We believe EIEE constitutes a type of epileptic encephalopaty which can be distinguished from other types of early onset epilepsy and should be included in the International Classification of Epilepsies with the West and Lennox-Gastaut syndromes. PMID- 7497180 TI - [Association of dystrophinopathy with facial-scapular-humeral dystrophy in the same family]. AB - We present the association of a distrophinopathy with a case of facioscupulohumeral dystrophy in two individuals belonging to the same family. The discrepancy in the seric creatinphosphokinase (CPK) of the two patients together with certain clinical data suggests the possibility that it is a question of two different processes. This impression was confirmed later through dystrophine analysis and genetic examination techniques. This case drew attention to the vital need today to insist on a combination of genetic examinations and dystrophine analysis when diagnosing muscular dystrophies, thus avoiding mistakes derived from diagnostic assumptions made on the basis of antecedents in the family involving neuromuscular disorders and the consequences that this might have regarding vital prognosis and advice to the family. This is the first case of coincidental association of these two processes within one family that has so far come to our knowledge. PMID- 7497181 TI - [An approach to hospital costs of cerebrovascular accidents]. AB - Taking into account that stroke is an important health problem, we have carried out this study in order to know the hospital cost of stroke in our Community. We have revised all the 210 patients admitted to the Neurological Division of the Hospital Virgen de los Lirios de Alcoy between the 1st October 1990 and the 30th September 1992. The population of the area during this period was 133,915 inhabitants. The number of days staying in the hospital, the number of CT and IMR performed, the number of days that each patient was rehabilitated and the cost generated by all the ambulance transfers, were recorded. The total cost was 80,472,883 pts [hospital-stay: 73,735,200 pts (91.6%); CT: 3,922,000 pts (4.9%); IRM: 550,000 pts (0.7%); transfers: 982,492 pts (1.2%); rehabilitation: 1,283,191 pts (1.6%)]. This means a cost of 383,205 pts/patient, 220,474 pts/day and 1.7 pts/inhabitant/day. PMID- 7497182 TI - [Neuromuscular gait disorders in pediatric age]. AB - Alterations in gait are a frequent cause of consultation in neuropaediatric units. They have several different causes. One subgroup of special interest concerns those which have their origin in a neuromuscular disease. We studied the experience of the Child Neurology Unit at the Hospital 12 October from 1980 to 1993. MATERIAL AND METHODS: between 1980 and 1993 a total of 176 were studied because they showed alterations in their gait caused by a neuromuscular disease. Frequency and clinical features were analysed (sex, age at the beginning of the deterioration, duration of the latter etc.). RESULTS AND CONCLUSIONS: the alterations in gait were separated into two groups: Group I: backward or non acquired gait. Group II: deterioration in gait. IIa: temporary. IIb: definitive. The most frequent cause in Group I was spinal muscular atrophy and in Group II, muscular dystrophy linked to sex. To sum up, muscular diseases are far more important than the neuropathic ones, except for Group IIa where acute demyelinating motor polyneuritis was to be found at the top of the list of frequent occurrences. PMID- 7497183 TI - [The management of acute cerebrovascular disorders at the Emergency Services at the Castellon General Hospital]. AB - OBJECTIVE: To analyse the policy for hospital admission in the case of the patients who come to the Emergency Ward with acute cerebrovascular disorders. METHOD: The study was based on a retrospective analysis of the release reports from the Emergency Ward of Castellon General Hospital in the course of 1991, in which stroke, or transitory ischemic attack (TIA) were diagnosed. RESULTS: 514 patients were attended to who went to the Emergency Ward because of cerebrovascular disorder. Of these, 57.2% were referred to hospital, with 42% being sent home. It was observed that 10.65% (n = 23) they did not accomplish the criteria for stroke. There was an overwhelming number of cases of TIA diagnosed (50.2%) in patients who were not admitted, contrasting with only 7.5% being selected for admission. We considered possible explanations for this. CONCLUSIONS: We observed that as far as making the decision to admit a patient with acute cerebrovascular disorder, priority for admission was given in those cases which showed the worst neurological deficiencies, and had a worse prognosis as a consequence. Similarly, the older age (average 76.5 years old) was a prominent factor as far as those who were not admitted. PMID- 7497184 TI - [Infantile spasms in children with Down's syndrome]. AB - Two hundred eighty six infants with Down syndrome have been studied. Infantile spasms have been identified in nine of them, in which background, EEG pattern and its evolution, modalities of treatment and its effects, neuroimaging and development course have been revised. None of these patients had either familiar or personal pathological antecedents. The pattern in the first EEG made was hypsarrhytmic in all cases except one which showed a multifocal paroxystical activity, with intermittent and bilateral bursts of spike-waves. The treatment first used was ACTH in four cases, valproate in three cases and phenobarbital in two cases (one of these associated with nitrazepam). The ACTH treatment was effective in seven infants, either was the first or the second choice. An infant in whom the first treatment with ACTH was not successful, responded to the association with valproate+clonazepam. One patient treated initially with phenobarbital+nitrazepam, having no response to different prescriptions, responded finally to the association of carbamacepin+vigabatrin. The first treatment with valproate or phenobarbital had no effect in all patients. The EEG pattern improved in all cases just after the treatment response. Normal tracing was found for a period of two months to three years. Cranial TC was performed to three infants showing one of them a discrete ventricular dilatation and periventricular calcifications that suggested tuberous sclerosis. It is important to point out that, although the good effect of therapy and EEG pattern normalization, the development is below what had been expected in children with Down syndrome. Behavioral problems have been found in seven (77.7%) of these children.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7497185 TI - [Neurological studies of sleep]. PMID- 7497186 TI - [Treatment of erectile dysfunction in multiple sclerosis]. PMID- 7497188 TI - [Stroke units]. PMID- 7497187 TI - [A cerebral accident: ictus is a medical emergency]. PMID- 7497189 TI - [Genetic factors predisposing to multiple sclerosis: association with HLA system]. PMID- 7497191 TI - [Prognostic indicators of neurological sequelae in perinatal asphyxia: state of the art]. PMID- 7497190 TI - [Neurological diseases and driving]. AB - Traffic accidents (TA) are, after heart disease, cancer and stroke, the fourth death cause among the general population. Although the number of AT caused by diseases-excluding alcoholism- seems to be reduced, interaction between organic pathology and functional ability increases the importance of this problem. This paper revises the literature on the relation between AT and specific neurological diseases: epilepsy, obstructive sleep apnea syndrome (SAS), stroke, dementia and Parkinson disease. Also, the problems and the role of the neurologist in assessing driving ability in patients with brain damage is analyzed, with special reference to the legal condition in Spain. The insufficiency of diagnostic labels as predictors of driving ability is stressed; the group of patients affected by these pathologies does not present greater TA risk than young drivers twice that of the general population. In the cases of epilepsy, SAS and ECV, which can cause episodic driving inability, defining recurrence probabilities and finding regulation formulas is the task of clinical epidemiologists and the regulative authorities. In the case of dementia, Parkinson disease and ECV, causing psychomotor performance deterioration, the basic problem, complicated by the presence of comorbility in these patients, is the development of valid clinical scales for driving ability assessment. The regulative authorities need simple measures which are often difficult to develop. Meanwhile, it is the task of the neurologist, as part of the therapeutic intervention during the medical encounter, to discuss driving risks with each patient. PMID- 7497192 TI - [Guidelines for the management of acute cerebrovascular disease in patients]. AB - One of the challenges currently faced by Neurologists is to finish with the therapeutic nihilism that affects management of cerebrovascular disorders. It is possible to reduce damage resulting from stroke if adequate treatment is applied shortly after the beginning of symptoms, and within the 'therapeutic window'. This would mean that these disorders might be considered as treatable medical emergencies. The purpose of this report is to provide a guide for management of cerebrovascular disease, both ischemic and haemorrhagic, suggesting a diagnostic procedure, together with general and specific recommendations for the care of these patients, in order to avoid complications and improve outcome. PMID- 7497193 TI - [Non-valvular atrial fibrillation and cerebral infarction]. AB - Non-valvular auricular fibrillation (NVAF) is linked to a high rate of cerebral embolism (CE), being one of the main causes of cerebral infarction in elderly people. The real risk of these patients suffering a cerebral embolism depends on various factors such as the type of fibrillation (chronic as opposed to paroxistic) and on the coexistence or not of other cerebrovascular risk factors. In recent years five random essays have been published on the primary prophylaxis of CE in patients with NVAF (AFASAK, SPAF, BAATAF, CAFA and SPINAF) and one on its secondary prophylaxis (EAFT), which have evaluated the efficiency of oral anticoagulation, acetilsalicilic acid with placebo. The results indicate that oral anticoagulation is efficient and safe both in primary and secondary prophylaxis. Use of acetilsalicilic acid is safe, but less efficient, although it is easier to administer and more economical, and would be indicated for patients with a contraindication for anticoagulation or who are not compliant to therapy. Currently we need to determine which factors identify the patient subgroups with high risk NVAF both as regards the appearance of CE and that of haemorrhagic complications for anticoagulation. The identification of these subgroups would probably make it possible to give a more exact profile of the right treatment for these patients. PMID- 7497194 TI - [Stroke units: modality, structure, organization and utilization]. PMID- 7497195 TI - [The stroke unit a the Clinical Hospital in Madrid]. AB - The outcome in 195 patients admitted in a non-intensive stroke unit was compared to that in 198 patients admitted in general wards during a 12 months period. At admission, both groups were similar in prognostic indicators (age, sex, previous history, level of consciousness and motor impairment). Short-term mortality rates was reduced among patients admitted in stroke unit (p < 0.001). This difference was independent of patient's age, previous history and neurological impairment on admission. The length of stay in the hospital was also reduced (p < 0.001). Among survivors, functional outcome at discharge was better in those admitted in the stroke unit, with a greater proportion of patients independent for activities of daily living. We conclude that stroke unit in an efficient strategy that improves the clinical outcome of patients with acute stroke. PMID- 7497196 TI - [A comparative study of mortality and infectious complications in patients admitted to neurological and general wards]. AB - We analyse the outcome of patients with cerebral infarction depending the admission in neurological department or in general wards. We find an improvement of the changes in Toronto, Mathew and Barthel scales and a reduction in the mortality in patients admitted in Neurological department. The improvement is related with a reduction in infectious complications observed in the Neurological ward. Our data suggest that the place of the patient admission in the hospital may influence the prognosis of stroke patients and should be consider in patients with cerebrovascular disease. PMID- 7497197 TI - [Infirmary and cerebrovascular units]. AB - The cerebrovascular units have tried to reduce the death and morbility rates of patients suffering from cerebrovascular disorders. Only highly-trained personnel work on these units, with special emphasis being given to the role of the infirmary. In the current report, the different aspects connected with this role are examined. Stress is placed on the plans for health care, recommendation in the use of monitorisation scales, and activities regarding different altered patterns of health applied to cerebrovascular disorders. PMID- 7497199 TI - [Knowledge of the neurologist's role in the care, education and research of cerebrovascular pathology]. PMID- 7497198 TI - A formal overview of stroke unit trials. AB - We carried out a formal overview of the ten trials which compared the outcome of patients managed within a stroke unit with those managed in general wards. Care in a stroke unit was associated with an odds reduction for early mortality (median follow up 3 months) of 28% (95% confidence interval 8-44%; 2 p < 0.01), which was largely sustained (odds reduction 21% CI 1-37%; 2p < 0.05) at final review (median follow up 1 year). The odds reduction for a poor outcome (death or institutionalisation) at final review was 34% (95% CI 19-47%; 2p < 0.001). The mean length of stay in stroke units ranged from 61% to 133% (pooled result 96%) of that in general wards. Stroke patients managed within specialist units are more likely to be alive and living at home a year after the stroke than those managed in general wards. Stroke unit care does not apparently increase the time spent in hospital. PMID- 7497200 TI - [Moya-moya disease in a child with multiple malformations]. AB - Moya-Moya disease is an obstruction of the internal carotids and of the afferent and efferent channels of Willis's polygon, which causes a collateral circulation, responsible for the typical angiographic image of a "puff of smoke" (Moya-Moya, in Japanese). Its etiology is unknown, and it might be congenital or acquired. It has been associated with congenital diseases such as Neurofibromatosis and disease of the falciform cells, among others. It usually appears as an alternating hemiplegia. We report here on the case of a 4-year-old boy with alternating hemiplegia (initially on the left, later on the right and then back on the left) and angiographic alterations which are characteristic of Moya-Moya. This patient had been examined in our hospital in the neonatal period as he had malformative stigmas, basically consistent with a flat angioma on the forehead, upper eyelids, and base of the nose, a previous wide fontanella which expanded to the whole of the methopic suture, aplasia, forehead level skin and abnormal implantation of the skin in that region. What draws attention in this case is the association of middle line craniofacial lesions (angioma, skin aplasia) and the later development of Moya-Moya. PMID- 7497201 TI - [Hair disorders and neurological impairments (neurotricosis): three new cases]. AB - Three children with hair-shaft abnormalities and neurological disorders are reported. The first case is a young girl with monilethrix and Friedreich's ataxia that presented IgG and IgA deficiency, without telangiectasis. The second patient is a young girl with monilethrix and HMSN I. The third patient is a boy with a peculiar face, hypotrichosis and pili torti; he was mentally retarded and had epileptic seizures. His diagnosis was compatible with Noonan's syndrome or with a cardio-facio-cutaneous syndrome, because of the existence of hyperkeratosis and the neurological features. It is our aim to emphasize the heterogeneity of neurotrichosis, whose limits are not well established and whose manifestations are shared by some of them. PMID- 7497203 TI - [Wallenberg syndrome in childhood secondary to the striking by lightening]. AB - Wallenberg's Syndrome (WS) is exceptional in childhood. We present a case of a 10 year-old girl with a WS caused by an embolism in the right lateral portion of the medulla secondary to an electrofulguration, but with an unusual progressive outcome, developing clinical signs of medial medullar injury, leading even into a respiratory failure 30 days after admittance. She only became stable after surgical lesion decompression. There was no evidence of vascular malformation, finding only a great oedema. The pathology only showed changes compatible with brain infarction. This case makes again clear the superiority of MRI upon CT-scan to diagnose a WS. We think that the unusual progression of the lesion was due to great medullar oedema found on surgery, involving the brain regional flow. PMID- 7497204 TI - [A case of a 56-year-old woman with pulmonary lesions and focal neurological deficits]. PMID- 7497202 TI - [MELAS syndrome: clinical, pathological and neuroimaging study]. AB - The form of presentation of a new case of Melas Syndrome is described, together with a pathological and neuroimage study, including clinical development over a 3 year period. The usefulness of MR should be underlined here, given clinical doubts, and also normality in the EMG early phases of and the association with obstructive hypertrophic miocardiopathy. PMID- 7497205 TI - [Descriptive epidemiology of transitory ischemic attack: a study in Muro d'Alcoi]. AB - A door-to-door survey of transient ischemic attack was conducted in Muro de Alcoi using a randomized sample. We randomly choose 646 people of the 4.525 included in the census. The prevalence ratio found for TIA and stroke were 12.3 per 1,000 people (95% CI 3.9-20.7). The age-specific prevalence rates for male and for women were 12.4 per 1,000 (95% CI 0.3-24.5) and 12.3 per 1,000 (95% CI 0.4-24.2) respectively. The work suggests that methodology of ascertainment strongly influences in the obtained rates, and reinforces the data of our previous whole population work in the same area. PMID- 7497207 TI - [The usage of calcium channel blockers in acute cerebral ischemia]. PMID- 7497208 TI - [The rupture of the parasellar dermoid cyst: evaluation with CT and MRI]. PMID- 7497206 TI - [Descriptive epidemiology of risk factors in cerebrovascular disorders:a study in Muro d'Alcoi]. AB - We evaluated cerebrovascular risk factors and the values of cholesterol and triglicerides in the survey in Muro d'Alcoi using a door-to-door randomised interview. We found 14.9% hypertensives, 5.5% persons with diabetes, 1.2% with heart diseases and 42.1 persons with tabaccoism into the 646 persons evaluated. The Serum Cholesterol levels were high and we observed that 62% of males and 63.8% of females had values higher than 200 mg/dl. Our study showed that 24.8% of males and 12% of females presented values of triglicerides higher than 150 mg/dl. The data of risk factors were comparable with the findings described in Beniarres, Lorcha and Planes, located in the same healthy area. PMID- 7497211 TI - [Acute alcohol proximal (femoral) neuropathy]. PMID- 7497210 TI - [Infantile spasms in a patient with 18p monosomy]. PMID- 7497209 TI - [Neurinoma of the facial nerve in the stylomastoid foramen: evaluation with CT and MRI]. PMID- 7497212 TI - [Hemifacial spasm secondary to Lyme disease]. PMID- 7497213 TI - [Meningitis caused by Klebsiella pneumoniae]. PMID- 7497215 TI - [Should the stroke patients be admitted in intensive care units?]. PMID- 7497214 TI - [Should patients with acute cerebrovascular diseases be admitted to intensive care units?]. PMID- 7497216 TI - [Should patients with acute cerebrovascular disorders be admitted to intensive care units?]. PMID- 7497217 TI - [Should patients with acute cerebrovascular disorders be admitted to intensive care units?]. PMID- 7497219 TI - [Should patients with acute cerebrovascular disorders be admitted to the intensive care units?]. PMID- 7497218 TI - [Should patients with acute cerebrovascular disorders be admitted to intensive care units?]. PMID- 7497220 TI - [The concept of neuropsychology]. PMID- 7497223 TI - [Should neurology residents have an educational rotation period at the local hospital?]. PMID- 7497222 TI - [Should neurology residents have an educational rotation period at the local hospital?]. PMID- 7497221 TI - [The concepts of neuropsychology]. PMID- 7497224 TI - [Should neurology residents have an educational rotation period at the local hospital?]. PMID- 7497225 TI - [Should neurological residents have an educational rotation period at the local hospital?]. PMID- 7497226 TI - [Should neurological residents have an educational rotation period at the local hospital?]. PMID- 7497227 TI - [Should neurological residents have an educational rotation period at the local hospital?]. PMID- 7497228 TI - [Should neurological residents have an educational rotation period at the local hospital?]. PMID- 7497229 TI - [A headache and ischemic infarction of middle cerebral artery: a prospective study of 100 patient]. AB - The frequency and clinical characteristics of headache were assessed in a prospective study carried out among 100 patients suffering from ischaemic infarct in the area of the middle cerebral artery. Headache was found in 28 out of the 100 patients studied and proved to be significantly more frequent in those infarcts located in the superficial branches (p < 0.001) and in the dominant hemisphere (p < 0.05). We found no relationship between the existence of headache and the type of infarct, its extension or level of neurological affectation. Severe headache preceded stroke in 28.5% of cases, was simultaneous in 57.1% and followed in 14.2%. The average intensity of headache was 2.14 +/- 1.69 headache units. It was bilateral in 46.4% of patients, generalized in 14.2%, unilateral on the side of the infarct in 28.5% and counterlateral in the remaining 10.7%. PMID- 7497230 TI - Lacunar stroke associated headache is not related to intracranial large vessel disease. AB - It was recently suggested that lacunar strokes presenting with headache may be due to large intracranial artery disease. From the stroke database of the Neurology department, Hospital Sta Maria, we retrieved all lacunar strokes who had investigations of the intracranial circulation. Forty seven of the 208 lacunar strokes registered had either angiography (5 cases) or transcranial doppler (42 cases). Twelve subjects noticed headache during stroke onset, but all had normal investigations of the intracranial circulation. In five patients transcranial Doppler showed ipsilesional increased peak systolic velocities and another subject had a middle cerebral artery branch occlusion. None of them reported headache. Headache associated with lacunar strokes is not related to intracranial artery disease. PMID- 7497231 TI - [The prevalence rate for cardiovascular diseases antecedents in patients with migraine]. AB - Throughout the many studies carried out on patients suffering from migraine, we find that a series of diseases or abnormalities with cardiovascular etiologies may be associated with the risk of migraine. In some studies published so far there is a greater prevalence of distinct cardiovascular diseases among those patients suffering from migraine. The shortage of case-control studies which might otherwise definitely establish a relationship between migraine and other cardiovascular diseases leads us to cautiously interpret the meaning of these associations. We studied family and personal history of cardiovascular disease in a group of patients with migraine, and we carried out a case-control study to try to establish the greatest risk of suffering from these diseases by our patients. We found that family antecedents of migraine with the next of kin was a risk factor for getting migraine and that Raynaud's disease was associated with a greater risk of suffering with this pathology. PMID- 7497232 TI - [Migraine and histamine: determining histidine in plasma and cerebrospinal fluid during migraine attacks]. AB - We studied the plasma levels and cerebrospinal fluid (CSF) levels in twenty-nine patients suffering from migraine without aura and in eighteen others with aura. The samples were obtained during migraine attacks. We compared the results obtained with those from twenty-seven control subjects. Histidine levels in plasma were 1.18 +/- 0.51 mg/dl for the control group, 1.75 +/- 0.39 mg/dl in those suffering from migraine without aura and 2.07 +/- 0.64 mg/dl in those cases of migraine with aura. In both plasma and in cerebrospinal fluid concentrations of histidine were greater in patients with migraine than in controls (p < 0.001). Histidine is an amino acid precursor of histamine and its abnormalities may be demonstrative of the participation of this amine in migraine attacks, possibly within the state of neuronal hyperexcitability. PMID- 7497233 TI - [Subcutaneous sumatriptan in the treatment of migraine attacks. An analysis of its long term efficaciousness and tolerance]. AB - We present the results of treatment with subcutaneous sumatriptan in migraine attacks. The study comprised forty-two patients suffering from migraine both with and without aura, with migraine attacks not susceptible to analgesics, non steroid anti-inflammatory drugs (NSAID), ergotics or else intolerance to the same. Two groups were independently analyzed, one consisting of ten patients who had menstrual migraine continually for twelve months, the other consisting of thirty-two patients suffering from migraine with and without aura for six months. We assessed the effectiveness of the drug (reduction in the intensity and duration of the attack, action, speed, recurrence) and tolerance (adverse effects). The effectiveness of sumatriptan in relieving headache was 75.9% (80% in the case of the menstrual migraine group and 71.8% in the case of the migraine with and without aura group). This effectiveness was maintained in a similar fashion by analyzing independently the first and last months of treatment. Adverse effects were noted in 38.7% of patients treated (40% for the menstrual migraine group, 37.5% for those with migraine with and without aura). The most frequent effects were pain at the point of injection, a feeling of general tiredness, nausea and a sensation of tension in the neck or chest. These effects were largely slight and short lived. No serious adverse effects were reported. A long term analysis carried out on the menstrual migraine group shows the efficacy of sumatriptan is kept up, with improved tolerance of the drug and a decrease in the number of negative side effects noted.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7497235 TI - [Evaluation of usefulness of the IHS criteria in the diagnosis of functional headaches in pediatric age]. AB - Following the criteria of the IHS, we studied 117 children (63 female and 54 male, mean age 9.35 +/- 1.65 years), selecting those classified as migraine (77) or tension-type headaches (30), total 107. We analysed the frequency of clinical manifestations with each type of headache (type and localization of pain, severity of the headache--intensity, duration and degree of disability--and associated symptoms) and their contribution to the differential diagnosis of each entity (evaluated as the presence--sensitivity--or absence--specificity--of each clinical manifestation in each group). Our study shows that throbbing headache (49%) and a unilateral location (31%) as well as nausea and vomiting (33.6%), although constituting diagnostic criteria of migraine, according to the IHS, have acceptable sensibility (91-95%), but only slight specificity (34-40%) for infantile migraine. On the other hand, we found that a greater intensity of headache (22%), and the presence of abdominal pain (15%) as associated symptom, although not considered diagnostic criteria, are also parameters of high sensitivity (100-92%) respectively, and also of greater specificity (30-70%) than the anterior criteria. We conclude that the criteria of IHS for the diagnosis of headache in infancy may be valid although some considerations are necessary. The new IHS criteria for the headache diagnosis in adults [1] may be useful in pediatric age, but the experience on this use is spare, consequently the increment on the use of this classification and the future changes, could produce a best operativity in the diagnosis of headache in childrens. PMID- 7497234 TI - [Four new cases and a review of the literature concerning the migraine with csf pleocytosis syndrome]. AB - The syndrome known as migraine with cerebrospinal fluid (CSF) pleocytosis is a clinical syndrome of controversial pathogenesis characterized by typical migraine headaches occurring in a fixed period of time along with lymphocyte pleocytosis in CSF which persists during intercritical periods. We present four new cases of this syndrome diagnosed in our service and we review those cases published in the literature in which there were no personal headache antecedents, attempting in this way to separate this syndrome from other types of migraine as included in the present classification. There also exists the possibility that it might be a question of a secondary process, the pathogenesis of which may play a part in the activation of the humoral immune system, given the frequent existence of immunogenic antecedents the previous days as well as that of a humoral reaction expressed as an increase in IgG levels in CSF with normal blood electrophoresis. PMID- 7497237 TI - [Migraine in childhood and adolescence: a retrospective review of hospital cases]. AB - A retrospective study of 101 cases of infantile migraine aged between 3 and 14 years is reported. Both sexes were affected equally, being common migraine the most frequent form. The immediate positive family history for migraine and underlying precipitating factors were identified in 66% and 88% of the cases respectively. The electroencephalographic picture displayed focal spike and wave or sharp and slow wave discharges in 19.1% of the cases. The evolution was favourable in 92% and there was no correlated with headache frequency or treatment approach. The better therapeutic response was obtained when underlying precipitating factors were removed. The most effective prophylactic drugs in our series were flunarizine, propanolol and dimetotiazine. We discuss the most relevant features of the migraine in the infancy. PMID- 7497238 TI - [Profile of the migrainous patients according to drug intakes]. AB - Migraine headache is a common, recurrent and often incapacitating disorder resulting in a substantial loss of work days and productivity, significant consumption of medicines, and considerable economic cost. We performed this study in order to determine the characteristic features of migrainous patients with regular or frequent intake of ergotamine preparations and non steroidal anti inflammatory drugs (NSAIDs) and the existence of significant differences between both groups. 186 patients were included in the study, 36.6% suffering from migraine without aura and 63.4% from migraine with aura. 24.7% were males and 75.3% females. Mean age of the sample was 35.6 +/- 12.9 years. With regard to medication used for immediate relief of headache, 74.7% patients reported an intake of analgesics and 25.3% ergotamine preparations. In patients with analgesic intake the mean age was significantly lower than in patients with ergotamine for treatment of migraine. When clinic features of these two headache groups were examined, the duration of migraine attacks in the analgesic group was significantly higher (p = 0.0014) than in the ergotamine group, and the severity of migraine was lower (p < 0.05) than in this group. Among associated symptoms, nausea and vomiting were significantly less frequent (p = 0.024) in the analgesic group than in the group of patients with ergotamine intake. PMID- 7497236 TI - [A retrospective study of infant headache]. AB - Headache either as an isolated syndrome or as part of a symptomatic grouping is a frequent reason for medical consultation or hospitalization during childhood and adolescence. We review 94 clinical histories of patients between three and thirteen years of age. Headache was the reason for being hospitalized in all cases. Our aim was to assess its incidence rate, epidemiology, clinical characteristics and etiology in addition to evaluating as to whether complementary examinations carried out during hospitalization were worthwhile. Among the most significant results were the following: age (73 patients were over seven years old, 77.6%), time elapsed for symptomatology to evolve (exactly or less than one week in 45% of cases); family history of migraine in 55 cases (58.5%). The most frequent accompanying symptoms were vomiting (38.2%), nausea (22.3%) and abdominal pain (19.1%). Physical exam was normal in 63 cases (67%) while sixteen patients (17%) had neurological focal signs and/or signs of endocranial hypertension (ECHT). Electroencephalography was performed on 94.6% of the patients and proved pathological in 22 cases (25%). Brain computerized tomography (CT) scan was carried out on 92.5% of the patients with space occupying lesions in 3.2% of the cases. The most frequent final diagnosis (52% of patients) was one of migraine. We did not find any patients with intracranial expansionary processes not showing signs of ECHT and/or neurological focalization, for which reason we doubt the profitability of the almost routine practice of carrying out brain CT scan on patients when severe headache is the sole symptom and where there are no specific findings during physical examination.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7497239 TI - [Intake patterns of analgesics in patients with headache who came to the neurologist]. AB - We have analysed the pattern of analgesic use in a group of patients that came for the first time to the neurologist. We have examined the first intention analgesic use in patients with headache, before the therapeutic intervention of the neurologist. During a month, we have followed a group of 40 patients. 20 of them were admitted as ambulatory patients at a Headache Unit at a Hospital and the other 20 were attended as outpatients by a general neurologist. 'Over-the counter' analgesics were the most commonly used (paracetamol, acetylsalicylic acid and combinations of them with other products such as caffeine). More than a half of the patients had consumed a combination of analgesics. At the time of the investigation, the mean of analgesic use were greater in the group attended at the Headache Unit than in the one attended by the general neurologist. This could depend on the fact that chronic daily headache was more frequent in the first group and that they were older than the other group. These findings could be in relation with a larger evolution of their headache. PMID- 7497240 TI - [The role of the age on the quality of life of patients older than 65 years of age with vertebrobasilar ischemia treated with nicardipine]. AB - OBJECTIVES: To establish the role of the age on quality of life of a group of normotensive patients, older than 65 years with vertebrobasilar ischemia, treated with nicardipine. To evaluate the influence of the Nicardipine side effects on the quality of life of these patients within general practice prescription limits. METHOD: An epidemiological cohort study was performed during six months, in several Basic Health Areas distributed all round Spain. Nottingham Health Profile (NHP) was used to evaluate quality of life and was applied at the beginning and at the end of the study. Nicardipine side effects were collected in every evaluation. RESULTS: 838 patients were included in the study, mean age 72.4 +/- 6.8 years, 337 (40.2%) were males and 501 (59.8%) females. When we compared quality of life according to the age, no significant differences were obtained (RR = 0.89; CI 95% 0.77-1.30). NHP global score +/- SD was initially 36.5 +/- 22.8 and 24.5 +/- 19.1 at the end of the study, this result was statistically significant (p < 0.001), which means a percentage of improvement of 33%. These differences were also significant in every NHP area. Side effects were not frequent and with no correlation with quality of life of patients (p = ns). CONCLUSIONS: Our data show that neither the age nor side effects due to nicardipine have influence on quality of life of normotensive patients, older than 65 years with vertebrovascular ischemia, treated with nicardipine. Quality of life assessment can be an useful tool for therapeutical evaluation in these patients. PMID- 7497242 TI - [Migraine with pleocytosis: another autoimmune disease?]. PMID- 7497241 TI - [Physiopathology of the migraine]. PMID- 7497243 TI - [History and humanities]. PMID- 7497244 TI - [Genetics of migraine]. PMID- 7497245 TI - [Basic mechanisms of the migraine physiopathology]. PMID- 7497247 TI - [Factors brought on by migraine]. AB - Migraine is a paroxysmic abnormality in which asymtomatic periods alternate with the appearance of attacks. Such attacks are the end result of a chain of events leading on to the acute clinical syndrome. Amongst those phenomena which occur in the days prior to the attack starting, the factors which bring such attacks on have been widely studied by a great number of researchers. Identifying these initiating factors is a fundamental preventive element and looking into the behavioural mechanisms of such factors could prove useful in clarifying the pathogenic mechanisms of migraine. In the present study we review most of the works which have sought to identify these factors concerning the development of attacks and to work out their behavioural patterns. PMID- 7497246 TI - [Epidemiology of migraine]. AB - Knowing the epidemiology of migraine is important for a host of reasons. Migraine is one of the most prevalent types of headache. In addition, it constitutes a pathology which due to its cyclic, recurring and highly incapacitating characteristics has enormous repercussions on the quality of life of those affected; it is responsible for the loss of an important number of working days and of production; responsible too for the vast consumption of medicaments and consequently represents a considerable economic cost. An epidemiological study, however, of its pathology poses countless difficulties owing to its characteristics and its association with risk factors. In this present work we review the obstacles to carrying out migraine epidemiological studies and we also comment upon many such studies found in the literature. In each section we present the data obtained in such studies as were performed by our working group. PMID- 7497248 TI - [Stress as a precipitating factor in migraine]. AB - Stress is the precipitating factor in migraine which is most commonly recognized by patients. There are many affected who describe headaches brought on by stressful situations and events, although they also speak of the onset of their attacks during the period of calm immediately after such moments of stress. There are however few objective works in the literature which study the relationship between stress and migraine. In the present work we review that literature which does exist concerning such a relationship. PMID- 7497249 TI - A neurologist's reflections on boxing. II. Acute and chronic clinical findings secondary to central nervous system damage. PMID- 7497250 TI - A neurologist's reflections on boxing. III. Vascular injuries. PMID- 7497251 TI - [A 54-year-old man with dorsal pain]. PMID- 7497252 TI - [The assessment of neurological care needs in the health sector of de San Juan de Alicante by key-informants model]. AB - BACKGROUND: Specific health care planning should be based on the care request according the most adjusted evaluations method. Data obtained from each geographic area may be most comparable with other from close areas. AIM: To determinate the neurological needs in the health area 16 of Valencian Community that includes the care corresponding to Hospital of San Juan de Alicante. MATERIAL AND METHOD: We conducted a survey among key informants using the primary care physicians of the area. The population of the healthy area is 144,781 people and includes 78 primary care physicians. We used the formula proposed by Kurtzke in 1982 in order to calculate the neurological time. RESULTS: We obtain a response rate of 82% the total number of neurological hours requested was 5,823.84 hours by year, which supposes a need of 8.82 neurologists by 100,000 people and nears a needs of 12.77 neurologists for the healthy area. These data are clearly higher that the obtained in other close areas using the same methodology (5 neurologists by 100,000 inh in Alcoi, 6 neurologists by 100,000 inh in the area corresponding to General Hospital of Alicante). The topics that require more than 100 hours by year are: migraine (1,205), lumbar backache (1,104), alcoholism (762), cranial trauma (442), deafness (285), stroke (272), seizures (181), disk herniation (166), cervical backache (140), zoster (162), psychosis (118), which supposes 83.12% of the whole neurological time. CONCLUSIONS: The data situated the area within a model of direct neurological care similar to that in the United States. PMID- 7497253 TI - [Prevalence of migraine in a population of university students]. AB - A group of universities not related with health sciences were interviewed using the questionnaire 'Alcoi 1992', based on the operational diagnostic criteria of the International Headache Society (IHS), to evaluate the prevalence of migraine. This questionnaire for the diagnosis of migraine was validated in 1993, showing a high sensibility and specificity for the diagnosis of migraine. Sensitivity, specificity, predictive value and chance-corrected agreement rate for the diagnosis of migraine was 100%, 94%, 90% (PVpos), 1008 (PVneg) and 0.71 respectively. The study group was composed by 96 students, men and women; the mean age of the group was 21 years old. The prevalence of headache, migraine, tensional headache and cluster headache was 95.4%; 7.58%; 12.2% and 1.52% cases respectively. The prevalence for male of headache, migraine, tensional headache and cluster headache was 92.5%, 12.5%, 15.9% and 2.5% cases respectively. The prevalence for female of headache, migraine, tensional headache and cluster headache was 100%; 0%; 5.5% and 0% cases respectively. The questionnaire appears like an useful, fast and easy method for the evaluation of diagnosis of migraine in populations groups. PMID- 7497254 TI - [Prevalence of migraine in the medical student population as determined by means of the 'Alcoi 1992' questionnaire]. AB - A group of medical students were interviewed using the 'Alcoi 1992' questionnaire based on the diagnostic criteria of the International Headache Society (IHS) in order to assess the prevalence and various characteristics of migraine. This questionnaire for headache diagnosis was approved in 1993, showing both high sensitivity and exactness in diagnosing migraine. Sensitivity, exactness, predictive value and agreement index of the questionnaire in migraine diagnosis were 100%, 94%, 90% (positive) or 100% (negative) and 0.71% respectively. The group was made up of 96 students of both sexes, with an average age of 23.3 years (standard deviation 1.87). A total of 92 students, 35 male and 57 female, reported headaches. The total prevalence rate for headache was 95.8%, the reliability interval (RI) being 76.7-114.9. In the case of female students, the prevalence rate for headache was 96.6% (RI 71.7-121.5) and in the case of male students it was 94.6% (RI 63-125.7). A total of 20 students, six male and 14 female, fulfilled the criteria for migraine. The overall prevalence rate for migraine was 20.8% (RI 11.9-29.7), the prevalence for females being 23.7% (RI 11.6-35.8) and that for males 16.2% (RI 3.7-28.7). The questionnaire would appear to be a useful, rapid and simple method in the assessment of migraine diagnostic. PMID- 7497256 TI - The basic pharmacology of gangliosides. PMID- 7497255 TI - [The scientific work of Dionisio Nieto]. PMID- 7497258 TI - [Clinical trials on ganglioside effectiveness]. PMID- 7497259 TI - [Guillain-Barre syndrome and gangliosides]. PMID- 7497257 TI - [Anti-gangliosides antibodies in neuromuscular's pathology]. PMID- 7497261 TI - [Mitochondrial diseases and cerebrovascular pathology]. PMID- 7497260 TI - [Neuroimaging in Canavan disease]. PMID- 7497262 TI - [Cerebrovascular manifestations of infectious diseases]. PMID- 7497264 TI - [Obstructive apnea syndrome during sleep and cerebrovascular disease]. PMID- 7497265 TI - [Metabolic changes and cerebrovascular disease]. PMID- 7497267 TI - [Cerebrovascular diseases and non-infectious inflammatory arteriopathies]. PMID- 7497266 TI - [Iatrogenic cerebrovascular disease]. PMID- 7497263 TI - [Psychiatric diseases and ictus]. PMID- 7497268 TI - [Non-inflammatory arteriopathies]. PMID- 7497269 TI - [Hematologic causes of vascular diseases of the central nervous system]. PMID- 7497270 TI - [Arterial hypertension and cerebrovascular disease]. PMID- 7497271 TI - [Cardiogenic cerebral embolism]. PMID- 7497272 TI - [Cerebral ischemia and respiratory diseases]. PMID- 7497273 TI - [Cerebrovascular complications in patients with cancer]. PMID- 7497274 TI - [Neurological complications after transplants: cerebrovascular complications]. PMID- 7497276 TI - [Cerebrovascular manifestations of systemic diseases: introduction]. PMID- 7497277 TI - [Cerebrovascular manifestations of gastrointestinal diseases]. PMID- 7497275 TI - [Ictus, pregnancy and contraception]. PMID- 7497278 TI - [Hepatic diseases and ictus]. PMID- 7497279 TI - [Ictus and cutaneous vascular syndromes]. PMID- 7497280 TI - [Analgesics and non-steroidal anti-inflammatory agents in the treatment of migraine]. PMID- 7497281 TI - [Ergotamine derivatives]. PMID- 7497282 TI - [Sumatriptan ++ in the treatment of migraine and cluster headache]. PMID- 7497283 TI - [Calcium antagonists in the prophylaxis of migraine]. PMID- 7497284 TI - [Adrenergic betablockers]. PMID- 7497285 TI - [Serotonin uptake inhibitors]. PMID- 7497286 TI - [Antidepressants and migraine]. PMID- 7497287 TI - [Pharmacological treatment of migraine: other treatments]. PMID- 7497288 TI - [Therapeutic strategies in migraine: introduction]. PMID- 7497289 TI - [Treatment of the acute migraine attack]. PMID- 7497290 TI - [Status migrainosus]. PMID- 7497292 TI - [Prevention of migrainous infarction]. PMID- 7497293 TI - [Therapeutic strategies in menstrual migraine]. PMID- 7497295 TI - [Therapeutic strategies in mixed or combined headache]. PMID- 7497296 TI - [Migraine and epilepsy]. PMID- 7497294 TI - [Migraine in childhood]. PMID- 7497291 TI - [Prophylaxis of migraine]. PMID- 7497297 TI - [The development of human cerebral cortex]. PMID- 7497298 TI - [Neurotransmitters and cerebral development: physiologic and physiopathologic aspects]. PMID- 7497299 TI - [The development of electrocardiogram]. PMID- 7497300 TI - [Neonatal seizures]. PMID- 7497301 TI - [Intraventricular hemorrhage in a newborn]. PMID- 7497302 TI - [Cerebral congenital hypotonia]. PMID- 7497303 TI - [Cerebral magnetic resonance angiography]. PMID- 7497304 TI - [Non-bacterial meningitis]. PMID- 7497305 TI - [Organic hyperactivity]. PMID- 7497306 TI - [Facial paralysis]. PMID- 7497307 TI - [Neurofibromatosis]. PMID- 7497308 TI - [Diagnosis of organic aciduria]. PMID- 7497309 TI - [Chorea and athetosis in childhood]. PMID- 7497310 TI - [Sydenham rheumatic chorea]. PMID- 7497311 TI - [Dystonia]. PMID- 7497312 TI - [Tremor and ataxia]. PMID- 7497314 TI - [Nystagmus and other anomalous movements of the eye]. PMID- 7497313 TI - [Tics: clinical spectrum]. PMID- 7497316 TI - [Mid-line cerebral tumors in children]. PMID- 7497315 TI - [Stereotypies]. PMID- 7497317 TI - [Neuroendoscopy]. PMID- 7497318 TI - [Congenital hydrocephalus]. PMID- 7497319 TI - [Histological and immunohistochemical analysis of dystrophin++ in muscular dystrophy of Duchenne and Becker type]. PMID- 7497321 TI - [Western blotting in the diagnosis of dystrophinopathies]. PMID- 7497320 TI - [Assistance and limitations of molecular genetics in the diagnosis of the Xp21 processes]. PMID- 7497323 TI - [Becker muscular dystrophy: clinical analysis, genetic and pathologic correlations]. PMID- 7497322 TI - [Duchenne muscular dystrophy: clinical analysis and prospects of genetic therapy]. PMID- 7497324 TI - [An infant with dizziness]. PMID- 7497325 TI - Infection control. PMID- 7497326 TI - Infection control. PMID- 7497330 TI - Your insurance and investment organization restructures for the future. PMID- 7497331 TI - Magnification systems in clinical dentistry. PMID- 7497327 TI - Ottawa discrimination case. PMID- 7497329 TI - Creating value for your staff. AB - Your staff are the agents of success in your practice. It is through their collective, quality efforts that your vision comes to fruition. Treat this investment in human resources with care and nourishment, and you'll be well on your way to establishing a thriving dental practice--one which can claim that not only its patients, but its staff as well, are its biggest fans. PMID- 7497332 TI - Advanced clinical technology--clinical tool or expensive toy? AB - There should no longer be any doubt in dentists' minds when it comes to the importance of technology in the day-to-day activities of the dental office. But there is, and this doubt must be addressed if the profession is to grow and prosper to its full potential. Denying the importance of technology--or putting on blinders--only leads to complacency, and clearly does not support our professions goal of providing the finest dental care possible. This article addresses the use of technology in a clinically-related manner, and attempts to provide the practitioner with valuable practice management tools to allow for a prosperous and efficient dental practice. PMID- 7497328 TI - I do not understand it. PMID- 7497333 TI - The law and ethics in dentistry: a recent Human Rights Tribunal judgment with further guidelines concerning dental treatment of HIV-positive patients. PMID- 7497334 TI - An innovative method of cushioning metal clamp jaws during rubber dam isolation. AB - Competent rubber dam use increases both operating speed and treatment quality. In many instances, however, the application of rubber dam clamps may cause immediate and/or postoperative discomfort, and some dentists continue to use this as a "justifiable" excuse for not employing rubber dam. This paper reviews the iatrogenic potential of metal rubber dam clamps, and introduces a new cushioning technique that makes use of light-cured provisional material. PMID- 7497335 TI - Prosthetic relining and dietary counselling in elderly women. AB - A reduced consumption of hard-textured fibrous foods may promote the development of digestive disturbances in more susceptible elderly people who wear prostheses having deficient masticatory function. The purpose of this study was to evaluate the effects of a program combining denture relining and dietary counselling on the masticatory performance, fibre intake, and prevalence of digestive symptoms in elderly women with low chewing efficiency. This program was provided to 55 women between 55 and 74 years of age. Chewing ability was assessed quantitatively with the Swallowing Threshold Test Index (STTI), and qualitatively according to the participants' experience in chewing five specific foods. Nutritional data were collected using a food frequency questionnaire. The mean value of the STTI was significantly increased at three weeks and at six to nine months after completion of relining by 37 and 31 per cent, respectively. Subjects included one more food in their diet at the end of the six- to nine-month period only. Fibre intake from vegetables was significantly increased by 18 per cent, while colonic and gastrointestinal symptoms were both reduced. Prosthetic relining increased the masticatory performance of elderly women, but not to a functional level. In addition, dietary counselling stimulated subjects to eat foods that had previously been avoided. These observations suggest that the improvement of chewing efficiency, combined with dietary counselling, could reduce the presence of digestive symptoms. PMID- 7497336 TI - An SEM examination of etched dentin and the structure of the hybrid layer. AB - The clinical requirements of dentin bonds are that they should be non-permeable to oral fluids, seal dentinal tubules, protect the pulp, and be long lasting and durable. Dentin bonding systems that use acidic agents to remove the smear layer are currently being used. Acid conditioning not only removes the smear layer, but also demineralizes the surface of the intertubular dentin and produces intratubular demineralization and funnelling. A dentin bond is produced when hydrophillic resin monomers infiltrate the dentinal tubules and collagen of the demineralized intertubular zone, producing a hybrid layer. The use of a critical point drying technique and SEM allows a clear visualization of the structure of the hybrid layer. This study showed that currently used hydrophillic resin monomers are unable to completely infiltrate the demineralized zone, and it is speculated that this failure could contribute to microleakage and influence the long-term durability of the bond. It is also apparent that these bonds depend on the mechanical investment of collagen by the infiltrating monomer. Since none of the unfilled resins tested seem capable of completely infiltrating the demineralized collagenous zone, the degree of demineralization produced by the commercial acid concentrations in current use is questioned. More dilute acids than those available commercially are shown to reduce both the degree and depth of demineralization, and we suggest that the resultant thinner layer may lend itself to more complete resin infiltration of the collagen. PMID- 7497337 TI - [Impacted bicuspids--surgical and orthodontic treatment: the Laval experience]. AB - It happens that maxillary cuspids do not follow their normal path of eruption and remain impacted. We have reviewed the files of patients presenting with this anomaly and determined which surgical and orthodontic treatments they have received, with the help of a removable appliance. PMID- 7497339 TI - Global structure-acute toxicity relationships for mice using structural parameter ratios: new approach to molecular design and screening. AB - A method for a preliminary survey of the relationship between molecular structure and performance was described using 1506 random data of structure-acute toxicity for mice (intravenously dosed). The structural patterns of the weakest toxic structures (111) were extracted from the data and the patterns discriminated for 64.2% of the other structures (1395). As for the 826 structures of strongest toxicity, 78.3% were discriminated by these structural patterns. These results were obtained by using structural parameter ratios to describe the structural patterns and the exhaustive elimination process to select the best parameter ratio from many candidates. The results were summarized in the form of a chart which can be used for practical screening for the weakest toxic structures. PMID- 7497338 TI - Quantitative structure-activity study of the toxicity of benzonitriles to the ciliate Tetrahymena pyriformis. AB - The toxicities of 34 benzonitriles to Tetrahymena pyriformis have been measured. Structure-activity relationships indicate that for these compounds different mechanisms of toxic action are taking place dependent on the nature of the substituent. Benzonitrile itself, some halogenated and the toluene derivatives model as non-polar narcotics; more polar substituents model well as polar narcotics; whilst the nitro and aldehyde substituted benzonitriles, and compounds that may be metabolised to benzoquinone are shown to exhibit considerable excess toxicity and, thus, the probability is that they are acting by a specific mechanism of action. After the removal of two outliers, QSAR analysis reveals a significant three parameter equation, and confirms the importance of hydrophobicity and descriptors of reactivity for the comprehension and the prediction of the toxicity of the benzonitriles. PMID- 7497340 TI - Structure-toxicity relationships for alkanones and alkenones. AB - The relative toxicity (log IGC-1(50)) of 54 selected alkanones, both aliphatic and aromatic, as well as, alkenones and alkynones was evaluated in the static Tetrahymena pyriformis population growth assay. Excess toxicity, an indicator of bioreactivity, was associated only with the alpha-beta unsaturated alkenones and alkynones. Moreover, the alkynones were found to be more toxic than corresponding alkenones. A high quality 1-octanol/water partition coefficient (log Kow) dependent structure-toxicity relationship, log IGC-1(50) = 0.86 (log Kow) - 2.27; r2 = 0.955, was developed for alkanones. This QSAR represented the nonpolar narcosis mechanism of toxic action. Toxicity of alkenones was predicted by the highest-occupied-molecular-orbital energy (HOMO), log IGC-1(50) = -3.474 (HOMO) 35.357; r2 = 0.897, and the difference between HOMO and the lowest-unoccupied molecular-orbital energy (LUMO), log IGC-1(50) = -3.559 (HOMO-LUMO gap) - 36.106; r2 = 0.903. The alpha-beta unsaturated ketones are considered soft electrophiles. Moreover, the toxicity of the aliphatic alkanones and alkenones was predicted by log Kow and LUMO, log IGC-1(50) = 0.69 (log Kow) - 2.55 (LUMO) + 0.05; r2 = 0.852. PMID- 7497341 TI - [Qualitative research in Public Health]. PMID- 7497342 TI - [Methods of user participation in the evaluation and improvement of the quality of health services]. AB - Involvement of consumers, the external customers of health services, in quality of care evaluation activities is increasingly regarded as a requisite for quality improvement programs. There are many but not mutually exclusive approaches to incorporate consumer participation to quality improvement activities. This paper reviews the main characteristics, and potential usefulness and pitfalls of the more common approaches. Participation methods are grouped in two groups: those actively decided and initiated by consumers (i.e. complaints, choosing providers, consumers' associations initiatives, etc.), and those requested by providers (i.e. satisfaction and expectations surveys, patients' reports, etc.). Given that requested consumer involvement may constitute a key element in quality improvement programs, special attention is paid to critically analyzing its methodological implications. PMID- 7497344 TI - [Automobile driving by the aged]. AB - BACKGROUND: Owing to the progressive ageing of our society and also to the great importance that we attach to driving, the number of elderly drivers (over sixty five years old) is likely to increase significantly. It is also important to point out that during the last years it has been stated a bigger amount of traffic accidents among the members of this community. METHODS: In this article different aspects already published about aged drivers are revised. RESULTS: The reasons for this rise are both the greater number of old people with a driving licence and the deterioration in some capabilities which may appear at a certain age. CONCLUSIONS: The fact that Health Service professionals dealing with this specific group are of the first importance as far as their education in this area is emphasized. PMID- 7497343 TI - [Association of air pollution and mortality: a review of recent epidemiological studies]. AB - The aim of this study is to present a review of the work published over the past few years which deals with the relationship between air pollution (AP) and mortality from the epidemiological perspective, by examining the evolution of methodology used in the analysis along with changes in values considered as threshold levels. A literature review was carried out on of articles published in the MEDLINE and IME databases. From this search some 40 articles were selected. The review has been centred on those studies which examine the association of AP indicators and short term mortality. The design of the paper is usually retrospective and, in almost all cases, these are ecological studies. Study populations are, most of times, inhabitants of cities or particular geographic areas, with the usual analysis being a combination of time series approach and multivariate regression. Also taken into account in the majority of this papers, apart from time variables, are other confounding factors, especially meteorological conditions. The results of the papers reviewed are presented along with their principle methodological characteristics. A series of recent studies indicate that the relationship between AP, especially small particles, and mortality may exist below the guidelines established at National and International level. This study finishes with a discussion on the most outstanding aspects of the results of the papers reviewed. Also mentioned is the difficulty of determining AP indicators, and their levels, which are linked to mortality. PMID- 7497345 TI - [Health status, use of health services and reported morbidity: application of correspondence analysis]. AB - BACKGROUND: The study of the relationship between self-reported morbidity, health status and health care utilization presents methodological problems due to the variety of illnesses and medical conditions that one individual may report. In this article, correspondence analysis was use to analyse these relationships. METHODS: Data from the Spanish National Health Survey pertaining to the region of Catalonia was studied. Statistical analysis included multi-way correspondence analysis (MCA) followed by cluster analysis. RESULTS: The first factor extracted is defined by self-assessed health perception; the second, by limitation of activities, and the third is related to self-reported morbidity caused by chronic and acute health problems. Fourth and fifth factors, capture residual variability and missing values. Acute problems are more related to perception of poor health while chronic problems are related to perception of fair health. Also, it may be possible to distinguish self-reported morbidity due to relapses of chronic diseases from true acute health problems. Cluster analysis classified individuals into four groups: 1) healthy people; 2) people who assess their health as being poor and those with acute health problems; 3) people with chronic health problems, limited activity and a perception of fair health; and 4) missing values. CONCLUSIONS: Correspondence analysis is a useful tool when analyzing qualitative variables like those in a health survey. PMID- 7497346 TI - [Elaboration and evaluation of a fast detection system of mortality using Fourier analysis. Study of a value with maximal deviation]. AB - BACKGROUND: To calculate all causes weekly baseline mortality and an alarm threshold using Fourier analysis. To analyse the largest outlier detected in our series. METHODS: Madrid Undertaker database from October 1988 to September 1993, was used in the analysis. Orthogonal components were detected using Fourier analysis. Expected deaths and confidence limits were fitted using Serfling method. Alarm threshold was placed at a distance of 1.96 standard deviations above baseline. RESULTS: orthogonal frequencies with significant amplitudes corresponding to periods 26, 52, 104 and 156 weeks were detected. The second was the fundamental and its multiples were harmonics. In the time domain, baseline mortality showed a winter peak, declined to a summer plateau and presented its lowest level at the end of August. 21 weeks exceeded the alarm threshold. Of these 17 were related to influenza epidemics. The largest outlier corresponded to a heat wave in July 1991. CONCLUSIONS: A procedure similar to that proposed by R. E. Serfling (1963) to calculate baseline mortality, an alarm threshold and short term extrapolation using Madrid Undertaker database (C. Borrell, 1991) is presented. Madrid Undertaker database provide accurate and timely information about all causes mortality excess in Madrid. PMID- 7497348 TI - [Efficacy of an informative seminar on the certification of causes of death]. AB - BACKGROUND: Mortality statistics are of crucial importance in decision making from the point of view of Public Health Studies conducted on the reliability of such statistics have disclosed errors or biases, some of which may be avoided and/or reduced with information and training. The aim of the present work was to evaluate the efficacy of an informative seminar, received by pregraduate students, about the certification of causes of death in improving the formal quality of such statistics, thus increasing their reliability. METHODS: A total of 175 students in the 6th year of their medical studies participated voluntarily in the Seminar. The students first made 6 certifications of death by different causes, then received a theoretical session, and finally made a further 6 certifications of death in a new test. RESULTS: The results obtained point to an improvement in the set of indicators employed in the evaluation of the quality of the certifications of death. CONCLUSION: The informative seminar mode is proposed as a method to be used in the training of health personnel in mortality statistics. PMID- 7497347 TI - [Diffusion of hepatitis B among residents of an institution for handicapped children]. AB - BACKGROUND: To estimate the prevalence of serologic markers of hepatitis B (SMHB) among residents of an institution for social or mental handicapped children in Castellon, and its association with risk factors. METHODS: A cross-sectional study with determination of SMHB by radioimmunoassay. 98 of the 100 residents participate in the study. RESULTS: Prevalence of SMHB was 24.5% (95% confidence interval: 16.4%-34.1%). Seven residents were hepatitis B surface antigen positive with three hepatitis B e antigen positive. SMHB prevalence was 0 in infants, 18.8% in toddlers, 24.5% in schoolchildren, 11.1% in special education, and 76.9% in permanent residents. Logistic regression analysis found associations of SMHB prevalence with age (Odds ratio [OR] = 5.8, 95% CI: 1.1-29.0), old age at admission with protective character (OR = 0.6, 95% CI: 0.4-0.8), and duration of stay (OR = 1.2 95% CI: 1.0-1.3), adjusting by sex, mental deficiency, and hepatitis B (HB) determinants like surgery, blood transfusion, odontology, bites, and injections. CONCLUSIONS: A predominant transmission of HB in young ages is suggested. Vaccination against HB, hygienic measures, and periodic serologic studies of HB are essential to prevent this disease. PMID- 7497349 TI - [Study of a typhoid fever outbreak in Baiona (Pontevedra)]. AB - BACKGROUND: Despite of the fact of being a high endemic treat, the notification of a typhoid fever outbreak produces a very unusual epidemiological happening in our province. The uncommon of the sight and the importance of this process has made us focus on this study in order to understand its causes as well as to be able to establish some control according to them. METHODS: In order to do that a retrospective epidemiological case-control study was made. Information was gathered about people who drank water from a public fountain in the village of Lages (Baiona) 14 cases and 52 controls randomly by hazard among the population of Lages and Almirante Fontan. The age mean of the people who became ill was between 25.6 +/- 16.9 years old and those control were 43.0 +/- 18.4 (p = 0.003). Among the 14 with the illness, 3 (21.4%) were men and 11 (78.6%) were women. RESULTS: The most frequent symptom was the fever (100%), followed by the cephaly (85.7%). The final OR was 57 (IC 95%: 13.2 - 246.2), being in Almirante Fontan a little higher (OR: 77; IC 95%: 4.7 - 572.1) than in Lages (OR: 40; IC 95%: 4.5 - 357.8). Salmonella Typhi strains was isolated in a 77.8% of the cases (7 of 9) where the blood culture was made. The water bacteriological analysis study proved a fecal contamination, but Salmonella Typhi strains wasn't isolated. CONCLUSIONS: The epidemiological and analytical suspicions guide us towards the water from the above mentioned fountain as the possible responsible for the coming about of this outbreak. PMID- 7497350 TI - [Gastroenteritis outbreak associated with water consumption, possibly caused by Norwalk or Norwalk-like virus]. AB - BACKGROUND: It is described an acute gastroenteritis outbreak of probable hydric transmission in Ontinyent (Valencia). It was declared on the 31st of January, 1992 and affected 3541 people according to the declaration done by the sanitary services who attended the patients. The clinical situation was characterized by the presence of profuse and watery diarrhoea, nausea, vomiting, abdominal pain and fever or febricula. METHODS: A preliminary study of the cases has been done, the information being treated with the classical method of descriptive epidemiology and a later study of the transversal observation type by means of a telephone inquiry. RESULTS: The results show that this outbreak affected approximately to a 30% of the population, and show the relation between consumption of water from the municipal water system and the outbreak, as well as the existence of a control population not affected for receiving a different water supply. CONCLUSION: Once rejected the bacterial origin for the results of the copro-cultivation done the analysis of the clinical and epidemiological characteristics of the outbreak, show a total agreement with the criteria described by Kaplan to characterize acute gastroenteritis outbreaks because of Norwalk-like virus. PMID- 7497351 TI - Calcium ion influx during sporulation in the yeast Saccharomyces cerevisiae. AB - The changes in intra- and (or) extra-cellular concentrations of Ca2+, Mg2+, K+, and Na+ during sporulation of a MATa/MAT alpha diploid yeast of Saccharomyces cerevisiae were examined in a nutrition-deprived medium with potassium acetate. Among these, Ca2+ in external medium was preferentially incorporated into cells, and sporulation was induced when the magnitude of free Ca2+ gradient between cytosol [Ca2+]i and external medium [Ca2+]o reached more than 3 x 10(3) ([Ca2+]i/[Ca2+]o = 3.5 x 10(3)). The result indicated that the meiosis and (or) sporulation signal of the yeast S. cerevisiae was generated through elevated Ca2+ influx rather than release from the internal Ca2+ stores. PMID- 7497352 TI - New mini-Tn5 derivatives for insertion mutagenesis and genetic engineering in gram-negative bacteria. AB - Five mini-Tn5 derivatives encoding resistance to Km, Cm, Gm, Tc, and Sm, coupled with the polylinker of the pBluescriptII plasmid, were constructed. These derivatives are carried by an ampicillin-resistant plasmid that has a conditional origin of replication from plasmid R6K and origin of conjugal transfer from the broad host range plasmid RP4. The new vectors are smaller than those previously described and possess numerous unique restriction sites inside the minitransposons for gene cloning in addition to SfiI and NotI sites found in their predecessors. PMID- 7497354 TI - Accumulation of Golgi-specific mannosyltransferases in Candida albicans cells grown in the presence of brefeldin A. AB - The fungal metabolite brefeldin A (BFA) is known for its ability to block the secretory process in eukaryotic cells by interfering in the endoplasmic reticulum (ER) to Golgi membrane traffic, causing the disassembly of Golgi apparatus and redistribution of Golgi enzymes into the ER. In sensitive yeasts, underglycosylated forms of secretory proteins accumulate in the cytoplasm in the presence of BFA. We investigated whether the incomplete glycosylation of mannoproteins could be due to repression of the synthesis of Golgi-located terminal mannosyltransferases and whether the underglycosylated mannoproteins can be incorporated into the cell walls in Candida albicans. However, we found that the microsomal membranes isolated from the yeast cells grown in the presence of 14 micrograms.mL-1 of BFA had on average three times higher overall specific activity of mannan synthase than membranes from control cells. The increase in specific activity of mannan synthase was mainly due to accumulation of Golgi specific mannosyltransferases responsible for elongation of the O-glycosidically linked mannooligosaccharides and for the synthesis of the N-glycosidically linked mannan outer chain. As a consequence, the mannans synthesized in vitro from GDP [U-14C]mannose by the membranes from cells grown in the presence of BFA had longer O-glycosidically linked oligosaccharides and longer side-chains in the N glycosidically linked polymannose part of the molecule than mannans synthesized by membranes from the control cells. Contrary to results obtained in vitro, the structural features of cell wall mannans isolated from intact BFA-grown and from control cells were almost indistinguishable. PMID- 7497353 TI - Antimicrobial actions of fluoride for oral bacteria. AB - Fluoride is widely used as a highly effective anticaries agent. Although it is felt that its anticaries action is related mainly to effects on mineral phases of teeth and on the process of remineralization, fluoride also has important effects on the bacteria of dental plaque, which are responsible for the acidification of plaque that results in demineralization. The results of recent studies have shown that fluoride can affect bacterial metabolism through a set of actions with fundamentally different mechanisms. It can act directly as an enzyme inhibitor, for example for the glycolytic enzyme enolase, which is inhibited in a quasi irreversible manner. Direct action seems also to occur in inhibition of heme based peroxidases with binding of fluoride to heme. The flavin-based peroxidases of many oral bacteria are insensitive to fluoride. Another mode of action involves formation of metal-fluoride complexes, most commonly AlF4-. These complexes are responsible for fluoride inhibition of proton-translocating F ATPases and are thought to act by mimicking phosphate to form complexes with ADP at reaction centers of the enzymes. However, the actions of fluoride that are most pertinent to reducing the cariogenicity of dental plaque are those related to its weak-acid character. Fluoride acts to enhance membrane permeabilities to protons and compromises the functioning of F-ATPases in exporting protons, thereby inducing cytoplasmic acidification and acid inhibition of glycolytic enzymes. Basically, fluoride acts to reduce the acid tolerance of the bacteria. It is most effective at acid pH values. In the acidic conditions of cariogenic plaque, fluoride at levels as low as 0.1 mM can cause complete arrest of glycolysis by intact cells of Streptococcus mutans. Overall, the anticaries actions of fluoride appear to be complex, involving effects both on bacteria and on mineral phases. The antibacterial actions of fluoride appear themselves to be complex but to be dominated by weak-acid effects. PMID- 7497355 TI - Effect of glucose on the superoxide dismutase production in fungal strain Humicola lutea. AB - The effect of glucose on superoxide dismutase (SOD) activity produced in Humicola lutea 110 was determined. The relatively high glucose concentration in the medium did not repress SOD levels in the cells. Glycerol, a nonfermentable carbon source, caused a slight stimulation of SOD synthesis. Furthermore, the specific rates of enzyme production in the medium with different glucose concentration showed an insignificant difference. Cyclic AMP had no effect on SOD levels. The shift in metabolism as glucose was depleted resulted in an increase in the rate of synthesis of both isocitric dehydrogenase and SOD. Pentachlorophenol and paraquat, which cause the production of superoxide radicals, caused an increase in SOD activity. These results led us to conclude that it is superoxide ion rather than glucose that is controlling SOD levels. PMID- 7497357 TI - To reuse or not to reuse? PMID- 7497356 TI - Pseudomonas aeruginosa strain MA01 aerobically metabolizes the aminodinitrotoluenes produced by 2,4,6-trinitrotoluene nitro group reduction. AB - Many microbes reduce the nitro substituents of 2,4,6-trinitrotoluene (TNT), producing aminodinitrotoluenes (ADNTs). These compounds are recalcitrant to further breakdown and are acutely toxic. In a search for organisms capable of metabolizing ADNTs, a bacterial strain was isolated for the ability to use 2 aminobenzoate (anthranilate) as sole C-source. This isolate, Pseudomonas aeruginosa MA01, metabolized TNT by first reducing one nitro group to form either 2-amino-4,6-dinitrotoluene (2ADNT) or 4-amino-2,6-dinitrotoluene (4ADNT). However, strain MA01 was distinct from other TNT-reducing organisms in that it transformed these compounds into highly polar metabolites through an O2-dependent process. Strain MA01 was able to cometabolize TNT, 2ADNT, and 4ADNT in the presence of a variety of carbon and energy sources. During aerobic cometabolism with succinate, 45% of uniformly ring-labeled [14C]TNT was transformed to highly polar compounds. Aerobic cometabolism of purified [14C]2ADNT and [14C]4ADNT with succinate as C-source produced similar amounts of these polar metabolites. During O2-limited cometabolism with succinate as C-source and nitrate as electron acceptor, less than 8% of the [14C]TNT was transformed to polar metabolites. Purified 2,6-diamino-4-nitrotoluene was not metabolized, and while 2,4-diamino-6 nitrotoluene was acetylated, the product (N-acetyl-2,4-diamino-6-nitrotoluene) was not further metabolized. Therefore, strain MA01 metabolized TNT by oxidation of the ADNTs and not by reduction the remaining nitro groups on the ADNTs. PMID- 7497358 TI - Ampullary tumour. PMID- 7497359 TI - Radiology for the surgeon. Human immunodeficiency virus cholangiopathy caused by cytomegalovirus or Cryptosporidium. PMID- 7497360 TI - Natural history of strictureplasty in Crohn's disease: 9-year experience. AB - OBJECTIVE: To study the short- and long-term outcomes in patients with Crohn's disease who have undergone strictureplasty. DESIGN: A retrospective review with a prospective follow-up (mean 54.4 months [range from 4 to 108 months]). SETTING: The Inflammatory Bowel Disease Centre at Mount Sinai Hospital in Toronto. PATIENTS: Forty-three patients (29 men, 14 women) who underwent 154 strictureplasties for Crohn's disease. The mean age of the patients was 32.5 years (range from 17 to 55 years). INTERVENTION: Strictureplasty by either the Heineke-Mikulicz (145 strictureplasties) or the Finney (9 strictureplasties) technique in the duodenum, small intestine and at the site of the previous anastomosis. MAIN OUTCOME MEASURES: Factors studied for symptomatic recurrence included the type of procedure previously performed, the type of strictureplasty, the number of previous operations and the site of the disease. RESULTS: There were no deaths. There was one documented leak. Twenty-six patients remained symptom free during the follow-up period. Fourteen patients required reoperation for progressive obstructive disease. None of the differences in the variables studied was statistically significant when related to the symptomatic recurrence rate. However, only 2 of 11 patients who had strictureplasty as the only procedure have required reoperation. CONCLUSIONS: Strictureplasty is a safe and useful procedure in the management of extensive obstructive Crohn's disease. Strictureplasty does not seem to alter the natural history of the disease. PMID- 7497361 TI - Intra-arterial infusion of urokinase for acute, critical ischemia in the lower limb. AB - OBJECTIVE: To assess the value of intra-arterial urokinase infusion in the management of acute, critical ischemia of the lower limb. DESIGN: A prospective study. SETTING: A vascular surgery department within a university hospital. PATIENTS: Twenty-five consecutive patients with acute, critical ischemia of the lower limb, excluding those requiring immediate surgical intervention. Follow-up ranged from 1 to 18 months. INTERVENTION: Angiography followed by intra-arterial infusion of urokinase. MAIN OUTCOME MEASURES: Angiographic and clinical evidence of clot lysis and limb reperfusion, any surgical procedures required and final clinical outcome. RESULTS: Urokinase was technically successful in lysing clot in 19 patients: 7 required no further treatment; in 8 an underlying lesion was identified and repaired by either percutaneous angioplasty or surgery; in the remaining 4 patients, although the clot was lysed, the limbs remained ischemic and, since reconstruction was considered impossible, amputation was required. Two patients improved clinically with little angiographic evidence of clot lysis. A total of nine patients required amputation, seven of these as a "primary" procedure after urokinase infusion. There were four episodes of significant morbidity but no deaths. CONCLUSION: Urokinase has a place in the management of acute vascular occlusion of the lower limb, not only in treating the occlusion but, equally importantly, in facilitating identification of lesions that require surgical intervention. PMID- 7497362 TI - Cost-effectiveness and efficacy of an on-site homograft heart-valve bank. AB - OBJECTIVE: To compare the performance and cost-effectiveness of cryopreserved homograft cardiac valves prepared on site to valves prepared commercially. DESIGN: A review of all patients who received homograft heart valves between January 1990 and December 1993, with cost-effectiveness analysis. Follow-up ranged from 1 to 37 months (mean 12 months). SETTING: Tertiary-care adult and pediatric hospitals. PATIENTS: All consecutive patients receiving homograft heart valves in either the aortic or right ventricular outflow tract (RVOT) position since on-site preparation of cryopreserved cardiac valves began in 1990. Forty three patients received 47 homograft valve replacements: 18 in the aortic position and 29 in the RVOT position. No patients were lost to follow-up. MAIN OUTCOME MEASURES: Valve function as assessed by patient survival, actuarial freedom from reoperation and Doppler echocardiographic assessment of transvalvar gradients and valvar insufficiency. Cost-effectiveness as assessed by a formal evaluation of on-site costs compared with current prices for commercially prepared valves. RESULTS: There were four operative deaths but no late deaths. Four valves were removed. Freedom from reoperation at 3 years was 100% for aortic valve replacement and 85% for RVOT reconstruction. Echocardiographic follow-up of 34 of the remaining 39 patients showed mild or no insufficiency in 24 valves, moderate insufficiency in 8 valves and severe, but not clinically significant, insufficiency in 2 valves. The cost of on-site preparation of the valves was $1363 compared with $5040 for the commercially prepared valves, a cost saving for the group of $172,819. CONCLUSION: On-site preparation is an effective method of preparing cryopreserved cardiac valves and permits significant cost savings. PMID- 7497363 TI - Reuse of disposable laparoscopic instruments: a study of related surgical complications. AB - OBJECTIVE: To determine the rate of surgical complications related to reuse of disposable laparoscopic instruments. DESIGN: A review of all laparoscopic and thoracoscopic procedures done between August 1990 and January 1994 in which disposable laparoscopic instruments were reused. SETTING: Department of general surgery in a 461-bed teaching hospital. PATIENTS: Patients who underwent 874 laparoscopic and thoracoscopic procedures in which disposable laparoscopic instruments were reused according to validated reprocessing protocols. MAIN OUTCOME MEASURES: The incidence of superficial and deep wound infections and complications related to instrument malfunction. RESULTS: The combined rate for deep and superficial infections was 1.8%. No complications related to a disposable instrument malfunction were found. CONCLUSION: Disposable laparoscopic instruments may be safely reused under carefully monitored conditions and with strict guidelines. PMID- 7497364 TI - A Madreporic-surfaced femoral component in a canine total hip arthroplasty model: bone remodelling response at 6 and 24 months. AB - OBJECTIVE: To evaluate the bone-remodelling response at 6 and 24 months after total hip arthroplasty around a two-thirds Madreporic-surfaced femoral component in a canine model. DESIGN: A research study in dogs. Statistical analysis was by multivariate analysis of variance. SETTING: A musculoskeletal research laboratory in a university teaching hospital. MATERIAL: Ten mature mixed-breed dogs. INTERVENTION: Unilateral implantation of total hip arthroplasty in the right femur with a Madreporic-surfaced prosthesis. Studies were conducted at 6 months in five dogs and at 24 months in five dogs. In each case, the nonoperated femur was the study control. MAIN OUTCOME MEASURES: Fluorescence microscopy analysis on comparable sections at four levels of the femora, percent porosity computed by image analysis, cortical atrophy. RESULTS: Cancellous hypertrophy was most prominent at the junction of the beaded and smooth portions of the femoral stem. At 6 months the cortex demonstrated atrophy predominant in the anteromedial quadrant. Active bone turnover was present within the interstices of the prosthesis at 6 and 24 months. No significant cortical bone loss occurred between 6 and 24 months. Stress transfer to the bone-prosthesis interface and endosteal cortical "cancellization" appear to be linked phenomena. PMID- 7497365 TI - The inflammatory response to particulate wear debris in total hip arthroplasty. AB - OBJECTIVE: To identify the cell types and patterns of cell distribution associated with particulate wear debris in the membranes of patients who undergo revision of hip arthroplasty. DESIGN: Case series. SETTING: Three university affiliated hospitals. PATIENTS: Twenty-nine patients who underwent revision of a hip arthroplasty for reasons other than infection. Eight patients were excluded because loosening of the prosthesis was found to be associated with infection, leaving 21 patients, 18 with aseptic loosening and 3 with recurrent dislocation. INTERVENTION: Retrieval of inflammatory membranes from loosened total hip prostheses. MAIN OUTCOME MEASURES: Results of aerobic and anaerobic tissue cultures, number of macrophages, T cells and B cells, and presence and histologic pattern of particulate debris in the membranes. RESULTS: None of the tissue cultures grew any organisms. All membranes contained T cells and macrophages, but B cells were present in only three patients at a low level. Particulate debris, surrounded by macrophages and foreign-body giant cells, was present in all membranes. Three histologic patterns were noted: type I--mostly fibrous tissue with cells and macrophages widely scattered through membrane (3 membranes); type II--fibrous membrane with a significant number of T cells, macrophages and foreign-body giant cells and well-defined foreign-body granulomas (7 membranes); type III--diffusely scattered T cells, ill-defined granulomas, "foamy" macrophages and fewer foreign-body giant cells than in type II (11 membranes). CONCLUSIONS: These findings document the key cellular elements involved in cell mediated immunity in association with particulate wear debris in aseptically loosened total hip arthroplasty membranes. T-cell amplification of the innate macrophage inflammatory reaction to particulate debris may play a role in the mechanism of particle clearance. PMID- 7497366 TI - Fixation of intertrochanteric hip fractures: gamma nail versus dynamic hip screw. A randomized, prospective study. AB - OBJECTIVE: To compare the efficacy of the gamma nail (GN) to the dynamic hip screw (DHS) in the management of intertrochanteric hip fractures. DESIGN: Randomized, prospective clinical trial with a mean follow-up of 52 weeks (range from 11 to 82 weeks). SETTING: A university teaching hospital. PATIENTS: One hundred and one patients with 102 fractures: 49 fractures were treated with the DHS and 53 fractures were treated with the GN. INTERVENTIONS: Fracture fixation with the DHS or the GN. MAIN OUTCOME MEASURES: Comparison of duration of operation, blood loss, early and late complications, functional outcome and duration of hospital stay. RESULTS: There was no significant difference between the two groups with respect to intraoperative blood loss, days in hospital, time to union and eventual functional outcome. The length of the procedure and fluoroscopy time were longer for the GN group. CONCLUSIONS: Both the GN and the DHS can be used effectively for the treatment of intertrochanteric fractures. In this study the DHS was associated with a lower risk of local complications and should still be considered to be the implant of choice for patients with intertrochanteric fractures. PMID- 7497367 TI - The effect of intramedullary cement on intercalary allograft reconstruction of bone defects after tumour resection: a pilot study. AB - OBJECTIVE: A pilot study to evaluate the surgical and functional results of resection and reconstruction of diaphyseal bone tumours with cemented and uncemented intercalary allografts. DESIGN: A retrospective case series. SETTING: A quaternary care unit within a teaching hospital. PATIENTS: Twenty-two patients with diaphyseal bone tumours treated between 1988 and 1992, with mean follow-up of 38 months (range from 20 to 70 months). INTERVENTIONS: Diagnostic biopsy in all patients. Preoperative and postoperative chemotherapy in patients with high grade sarcoma. En-bloc excision of all tumours. Reconstruction of the defect with cemented (15 patients) or uncemented (7 patients) intercalary allografts. MAIN OUTCOME MEASURES: Nonunion at one or both osteosynthesis sites, death from systemic disease, amputation. RESULTS: Six of the 22 patients died of systemic disease with the graft intact, and 3 subsequently had amputation for either local recurrence (2 patients) or failure of free tissue transfer (1 patient), leaving 13 patients alive with the allograft in situ at an average of 36 months (range from 24 to 64 months). There were no revision procedures to remove or replace the original allograft. Five patients had nonunion at one or more osteosynthesis sites on plain radiographs 9 months after resection. Four patients with nonunion were in the cemented group of 13 patients available for assessment at 9 months, and 1 was in the uncemented group of 5 patients available for assessment at 9 months. There were two graft fractures, both in the uncemented group. CONCLUSION: The results of this pilot study offer guarded support for the use of pressurized cement in allografts prior to reconstruction of intercalary defects. PMID- 7497368 TI - In-vitro strength of flexor-tendon repairs. AB - OBJECTIVE: To compare the standard Tajima technique for flexor-tendon repair of the hand with three new techniques with respect to strength and technical ease of the repair and bulk at the repair site. DESIGN: A randomized analysis of human cadaver flexor digitorum superficialis tendon repairs. SETTING: A musculoskeletal research laboratory at a university-affiliated health centre. MATERIAL: Thirty nine fresh-frozen cadaveric flexor digitorum superficialis tendons from index, long and ring fingers harvested and divided transversely. The tendons were randomly allocated for repair by the current standard technique or one of three new techniques. INTERVENTIONS: The standard Tajima (modified Kessler) technique, and the Halsted, Savage or Silfverskiold technique. MAIN OUTCOME MEASURES: The time to perform each repair, the cross-sectional dimension and the load to failure of the tendon repairs. RESULTS: The Savage repair tolerated the highest loads, followed by the Halsted, Silfverskiold and Tajima repairs. The Silfverskiold technique was the least time-consuming to perform, followed by the Tajima, Halsted and Savage repairs. All of the techniques increased the cross sectional dimensions of the tendon: the Silfverskiold repair by at least 50% and the Savage repair by over 100%. Although none of the techniques tested performed ideally, all three new techniques provided greater initial static strength than the standard Tajima method. CONCLUSION: Further in-vitro and in-vivo studies of these new techniques of flexor-tendon repair of the hand are necessary to optimize the treatment of these injuries. PMID- 7497369 TI - Lower extremity burns and Unna paste: can we decrease health care costs without compromising patient care? AB - OBJECTIVE: To compare an alternative treatment for lower extremity burns with the standard in-hospital treatment, in an attempt to shorten hospital stay. DESIGN: A case-control series. SETTING: A university-affiliated hospital. PATIENTS: All patients with a burn isolated to a lower extremity were treated over an 8-month period with split-thickness skin grafting (STSG), Unna paste dressing, immediate mobilization and early discharge. This group was compared with matched controls from the preceding 8 years treated with STSG, occlusive burn gauze dressing, bed rest and hospitalization. MAIN OUTCOME MEASURES: Duration of hospital stay and graft viability. RESULTS: Thirteen patients with an average wound size of 131 cm2 were treated with Unna paste and had a graft viability of greater than 95% and a burn-scar rating equivalent to that of patients treated with the earlier regimen. The duration of hospital stay decreased from a mean of 12.9 days to 1.4 days, with no complications. This translated into a saving of $10,350 per patient. CONCLUSIONS: This alternative treatment is safe, inexpensive and effective and is recommended as the treatment of choice for uncomplicated, noncircumferential lower extremity burns. PMID- 7497370 TI - Long-term results of penile venous ligation for corporeal venous occlusive dysfunction. AB - OBJECTIVE: To evaluate the long-term results of penile venous ligation in patients with erectile dysfunction due to venous incompetence of the corpora cavernosa. DESIGN: A retrospective case study with follow-up longer than 22 months. SETTING: A university teaching hospital. PATIENTS: Thirty-two men with penile corporeal dysfunction, ranging in age from 33 to 77 years (mean 45 years). The duration of impotence ranged from 8 to 66 months (mean 29 months). The results were evaluated by chart review and patient questionnaire. INTERVENTION: Ligation of the penile cavernosal and crural veins. MAIN OUTCOME MEASURES: Penile brachial index (PBI), peak flow rate in the cavernosal arteries after injection of papaverine, penile rigidity and maintenance of perfusion rate during cavernosometry, all measured preoperatively and compared postoperatively, and major venous leakage, defined as leakage into both the deep and the intermediate systems. RESULTS: Ten (31%) of the 32 patients had a good result (full rigid erection without adjuvant therapy postoperatively) and 9 (28%) showed improvement (full erection with intracorporeal injection). Of the 13 (41%) failures, 7 had temporary improvement, ranging from 6 to 20 months. Preoperative penile rigidity, PBI and peak blood-flow velocity were significantly higher in patients with good results than in those with poor results (68% versus 40%, p < 0.05). Major venous leakage occurred in 12 of 13 patients with poor results but in only 1 of the patients with good results. CONCLUSIONS: The success or failure of penile venous ligation should not be evaluated during the 1st year after the procedure. The presence of minor leakage and penile rigidity greater than 60% are good predictors of the success of the procedure. PMID- 7497371 TI - Moore hemiarthroplasty functioning for 33 years: a case report. AB - The Moore monopolar hemiarthroplasty is useful in the management of patients with femoral neck fractures, but long-term results may be associated with acetabular cartilage degeneration and hip pain. The authors report a case of long-term function of a Moore hemiarthroplasty. A 61-year-old woman sustained a fracture of the neck of the femur, which was treated initially with pinning. One year later, because the operation had failed, the patient underwent a Moore hemiarthroplasty. The prosthesis functioned well for 32 years, when she experienced increasing pain in the groin and a decreasing range of hip motion. Radiologic examination showed thinning of the acetabular cartilage so a total hip arthroplasty was performed. This case demonstrates that a unipolar arthroplasty is capable of long-term function, and with current improvements in stem fixation improved longevity can be expected. PMID- 7497372 TI - Compression of the deep branch of the peroneal nerve by the extensor hallucis brevis muscle: a variation of the anterior tarsal tunnel syndrome. AB - Anterior tarsal tunnel syndrome is a rare condition in which the deep peroneal nerve is trapped below the extensor retinaculum at the ankle. The authors report a variation of this condition: a 67-year-old woman who had anterior tarsal tunnel syndrome caused by entrapment of the deep peroneal nerve by the extensor hallucis brevis muscle. Conservative treatment was unsuccessful, but surgical decompression of the nerve provided immediate improvement, and by 2 weeks postoperatively she had no residual pain or paresthesia, although there was some numbness in the first web space. PMID- 7497373 TI - Peripancreatic retroperitoneal gas mimicking necrotizing pancreatitis after laparoscopic cholecystectomy. AB - The computed tomographic image of gas in the pancreas and peripancreatic retroperitoneum is strongly suggestive of necrotizing pancreatitis. The authors describe an 81-year-old woman who, 7 days after laparoscopic cholecystectomy, presented with retroperitoneal gas but did not have the clinical or biochemical features of necrotizing pancreatitis. Resolution of her low-grade fever and of the radiologic findings with conservative treatment and the absence of other causes for pneumoretroperitoneum suggest that the gas observed was related to the laparoscopic cholecystectomy. PMID- 7497374 TI - An alternative approach to the Cabrol technique of aortic root replacement. PMID- 7497375 TI - The ambiguous mammography report and needle-directed biopsies. PMID- 7497376 TI - The radiologist's contribution to journal articles. PMID- 7497377 TI - Radiopacity of glass: does the lead content matter? PMID- 7497378 TI - Mixed reaction to suicide of terminally ill woman. PMID- 7497379 TI - Mixed reaction to suicide of terminally ill woman. PMID- 7497380 TI - Mixed reaction to suicide of terminally ill woman. PMID- 7497381 TI - Dyspepsia: costs of endoscopy. PMID- 7497382 TI - Dyspepsia: costs of endoscopy. PMID- 7497383 TI - Allow physicians who commit sex offences to work, but away from patients. PMID- 7497385 TI - Setting priorities and selecting topics for clinical practice guidelines. AB - Setting priorities and selecting topics are important steps in guidelines development, but they have received relatively little attention to date. Responses from a survey of guidelines stakeholders in Canada suggest that the health burden of a clinical condition on the population is an important factor in priority setting. Economic considerations, cast as either costs of treatment to the health care system or the economic burden of illness to society, are given varying importance by different stakeholder groups. Drawing on the literature and the survey results, the authors propose a framework for priority setting. Important issues requiring consideration include the role of public and community participation, the need for and appropriate emphasis on quantitative data regarding current practice and its variation, and mechanisms to link guidelines to health-policy development and management of the health care system. PMID- 7497384 TI - Physician mobility: individual rights. PMID- 7497386 TI - Managing erotic feelings in the physician-patient relationship. AB - In spite of prohibitions against the sexual involvement of physicians with their patients, erotic feelings sometimes arise in physician-patient relationships. The authors suggest that physicians can protect themselves and their patients from the harm that results from sexual involvement by establishing behavioural limits for their professional relationships, responding to patients' sexual overtures in a firm but nonjudgemental manner, examining their own sexual feelings rationally, seeking consultation if necessary and terminating the relationship if sexual feelings are compromising patient care. The challenge for physicians is to acknowledge that sexual feelings can arise and to manage such feelings for the sake of their own and their patients' well-being. PMID- 7497387 TI - Making sense of shadows: Dr. James Third and the introduction of x-rays, 1896 to 1902. AB - The discovery of x-rays was announced by German physicist Wilhelm Conrad Rontgen in December 1895. This review of the introduction of the use of x-rays in Kingston, Ont., shows the rapidity of their adoption in Canadian medicine. By February 1896 "x-ray photographs" were being taken by Captain John Cochrane of the Royal Military College of Canada in Kingston. Initially a scientific and popular curiosity, the new rays were quickly applied to medicine, and by the fall of 1896 the Kingston General Hospital had acquired its own x-ray apparatus. The hospital superintendent, Dr. James Third, became a leading practitioner and promoter of radiographic diagnosis and radiation therapy. He published, in 1902, the first comprehensive review of the diagnostic and therapeutic uses of x-rays by a Canadian physician. Third's writings reveal his technical knowledge, his organized approach to the application of radiography to clinical medicine and his cautious attitude. Like other physicians who have witnessed the introduction of new diagnostic techniques, Third feared that the new technology would usurp the physician's clinical skills. PMID- 7497389 TI - HIV infection among women undergoing abortion in Montreal. AB - OBJECTIVE: To determine the seroprevalence and correlates of HIV infection in a subpopulation of women of childbearing age in Montreal. DESIGN: Anonymous unlinked seroprevalence study. SETTING: Pregnancy termination unit in a teaching hospital in Montreal. PARTICIPANTS: Women presenting for abortion from July 1989 to June 1993 who resided in Quebec and were not known to have HIV infection; 12,017 (99.6%) of 12,068 eligible women were included in the study. INTERVENTION: HIV antibody testing of serum left over from samples obtained for routine Rh typing; the same algorithm as for serodiagnostic testing, namely enzyme immunoassay (EIA) followed by confirmatory testing of repeatedly EIA-reactive samples, was used. OUTCOME MEASURES: HIV serostatus by age, marital status, region of residence (metropolitan Montreal versus other), country of birth and number of living children. RESULTS: Most (84.7%) of the subjects resided in metropolitan Montreal. The median age was 27.0 (range 13 to 50) years. The serum samples of 22 women were confirmed to be HIV positive, for an overall seroprevalence rate of 1.8 per 1000 (95% confidence interval 1.1 to 2.8). The seroprevalence rate did not vary significantly by age, marital status, region of residence or study year. However, it was strongly correlated with country of birth: Canada 0.16, Haiti 23.5, HIV-endemic countries other than Haiti 5.3 and non-HIV-endemic countries other than Canada 0.0 per 1000. The seroprevalence rate among women born in Haiti was 147 times higher than that among women born in Canada (p < 0.0001). Of the women born in Haiti the rate was 3.0 times greater among those who immigrated to Canada in 1985 or later than among those who immigrated earlier (p = 0.047). CONCLUSIONS: The results of this study indicate that the HIV seroprevalence rate among women in Montreal is strongly associated with country of birth, women born in HIV-endemic countries, especially Haiti, having the highest rate. These results will help in the development of policies regarding HIV antibody testing and prevention of HIV transmission in Quebec. PMID- 7497388 TI - Costs associated with gunshot wounds in Canada in 1991. AB - OBJECTIVE: To estimate the costs (in 1993 dollars) associated with gunshot wounds in Canada in 1991. DESIGN: Cost analysis using separate estimates of gunshot incidence rates and costs per incident for victims who died, those who survived and were admitted to hospital and those who survived and were treated and released from emergency departments. Estimates were based on costs for medical care, mental health care, public services (i.e., police investigation), productivity losses, funeral expenses, and individual and family pain, suffering and lost quality of life. SETTING: Canada. OUTCOME MEASURES: Costs per case, costs by type of incident (e.g., assault, suicide or unintentional shooting) and costs per capita. RESULTS: The total estimated cost associated with gunshot wounds was $6.6 billion. Of this, approximately $63 million was spent on medical and mental health care and $10 million on public services. Productivity losses exceeded $1.5 billion. The remaining cost represented the value attributed to pain, suffering and lost quality of life. Suicides and attempted suicides accounted for the bulk of the costs ($4.7 billion); homicides and assaults were the next most costly ($1.1 billion). The cost per survivor admitted to hospital was approximately $300,000; this amount included just over $29,000 for medical and mental health care. CONCLUSION: Costs associated with gunshot wounds were $235 per capita in Canada in 1991, as compared with $595 in the United States in 1992. The differences in these costs may be due to differences in gun availability in the two countries. This suggests that increased gun control may reduce Canada's costs, especially those related to suicide. PMID- 7497390 TI - Female family physicians in obstetrics: achieving personal balance. AB - OBJECTIVE: To describe the experiences of female family physicians who practise obstetrics in balancing professional obligations with personal and family needs, given the unique challenges that such practice poses for these physicians. DESIGN: Qualitative study. SETTING: Ontario. PARTICIPANTS: A purposefully selected sample of nine female family physicians who met the criteria of being married, having children and currently practising obstetrics. OUTCOME MEASURES: Experiences of female family physicians and their strategies in their personal, family and professional lives that enable them to continue practising obstetrics. RESULTS: All participants continued to practise obstetrics because of the pleasure they derived from it, despite the challenges of balancing the unpredictable demands of obstetrics with their personal and family needs. To continue in obstetrics, they needed to make changes in their lives, either through a gradual, evolutionary process or in response to a critical event. Alterations to work and family arrangements permitted them to meet the challenges and led to increased satisfaction. Changes included making supportive call-group arrangements, limiting work hours and the number of births attended and securing help with household duties. CONCLUSIONS: An in-depth examination, through the use of qualitative methods, showed the reasons why some female family physicians continue to practise obstetrics despite the stressful aspects of doing so. This knowledge may be useful for women who are residents or experienced clinicians and who are considering including obstetrics in their practice. PMID- 7497391 TI - Mycobacterium abscessus infection of a Norplant contraceptive implant site. AB - The authors report a case of Mycobacterium abscessus infection of a subdermal levonogestrel implant (Norplant) site. The infection lasted 12 weeks and was indolent, skin manifestations were low grade and difficult to detect. Culture of exudate samples showed that M. abscessus was the only causative agent. After the implant was removed the patient's arm healed uneventfully without antimycobacterial therapy. The authors recommend that if Gram staining of apparently infected material from an implant site does not reveal a causative organism, then cultures should be done for mycobacteria and fungi. Kinyoun staining for acid-fast bacteria and calcoflour-white staining for fungi should also be performed. The implant should be removed and the patient given antimicrobial therapy as indicated. The authors emphasize the need to be aware of the potential for M. abscessus infection of implant sites and stress that appropriate microbiologic culture procedures are essential for accurate diagnosis. PMID- 7497393 TI - Hantavirus pulmonary syndrome in Canada: update. PMID- 7497392 TI - Diagnostic imaging at its centennial: the past, the present and the future. AB - Since the discovery of x-rays by Wilhelm Conrad Rontgen 100 years ago, diagnostic imaging has profoundly influenced the practice of medicine. As a result of discoveries during this period, ultrasonography, nuclear imaging, computed tomography and magnetic resonance imaging, as well as conventional radiography, have assumed a major role in diagnostic medicine. In addition to their traditional role in diagnosis, imaging techniques are becoming an increasingly important factor in innovative treatment methods, and this role is likely to expand. In the current climate of rising health care costs, radiologists and other health care providers who use imaging must increasingly account to health care funders for the cost-effectiveness of imaging in relation to other diagnostic and interventional techniques. They must also assure minimum standards of quality and training, and determine the appropriate role for diagnostic imaging in health care systems of the future. PMID- 7497394 TI - Creation of first malaria vaccine raises troubling questions about "intellectual racism". Interview by Kirsteen MacLeod. AB - Some of the problems caused by malaria, which places a huge roadblock in front of economic progress in the Third World, may be solved by a new vaccine created by Dr. Manuel Patarroyo, a Columbian physician and researcher. "Imagine how things would be if Canadians had malaria," he says. "Episodes last 10 days, then there are 10 days of recovering. This leaves only 10 days each month in which to do some productive work. Then imagine killing the population of Toronto each year, and you can see the huge toll in terms of the number of yearly deaths globally from malaria." His discovery also raises the issue of "intellectual racism" because of criticism of Patarroyo's methods by Western scientists. Patarroyo, meanwhile, turned down a $60-million offer for his vaccine, and instead donated the patent to the World Health Organization. PMID- 7497395 TI - Psychologic scars remain 50 years after Dieppe raid, study of Canadian veterans finds. AB - A Toronto psychologist has conducted a 50-year follow-up study of the psychologic problems faced by Canadian troops who landed on the beaches of Dieppe, France, in 1942. Lynne Beal says many of the men have suffered in silence, never realizing that they were experiencing the widespread condition known as post-traumatic stress disorder. The veterans hope her work will help them win greater official recognition of their condition. PMID- 7497396 TI - The electronic medical-student exchange: a low-cost alternative to overseas electives. AB - The authors report on an international collaboration that uses the Internet for medical education. In addition to introducing the students to electronic communication, the project aims to further international collaboration and understanding and to emphasize the importance of a population perspective for health and disease. Students and professors in Canada, England and Hungary participated. PMID- 7497397 TI - Experts discuss cost-effective treatment of hypertension during world conference in Canada. AB - Even though an estimated 3.5 million Canadians are affected by hypertension, speakers at a World Conference on Hypertension Control questioned whether it is generally cost effective to treat younger men and women who have mild hypertension. Nonpharmacologic treatment via weight loss and lifestyle modification should be the first-line treatment, speakers stated. They looked at the basic principles for evaluating the economics of hypertension management and made recommendations on the cost effectiveness of treating various patient groups according to age and severity of their hypertension, and on the selection of diagnostic-evaluation procedures and antihypertensive medications. PMID- 7497398 TI - Counselling as death approaches: is the New Yorker really the best we can do? AB - An expert in palliative care has taken exception to a recent CMAJ article dealing with euthanasia. That column praised a magazine article in which a son explained why he and his family had helped their mother commit suicide in the face of a terminal illness. Dr. Balfour Mount says the article was misleading and offers no credible solutions to the issues surrounding palliative care. PMID- 7497399 TI - A 56-year-old medical calendar from McGill opens a fascinating window on medicine's past. AB - McGill University's Faculty of Medicine has a long history of excellence. An examination of one of its more mundane documents, the student calendar from 1939 40, provides an instructive look at the roots of this excellence and indicates how much things have changed and how much remains unchanged after more than half a century. PMID- 7497400 TI - Resolutions passed during the CMA's 1995 annual meeting. PMID- 7497401 TI - Innovative HIV/AIDS program launched in Ottawa. AB - Collaboration among 31 social and health care agencies and the provincial government has resulted in an innovative program for Ottawa-area patients with HIV infection or AIDS. The target group is the homeless and people with "unstable" housing who live in the city's downtown core, a group at high risk of contracting HIV. The education of family practitioners will be an important part of the program. PMID- 7497402 TI - Native healing traditions must be protected and preserved for future generations. AB - Toronto physician Mel Borins, who has travelled extensively to developing countries to examine traditional healing practices, recently visited elders at an Ontario Indian reserve to learn more about native healing practices. He is concerned that much native knowledge about the use of herbs and plants for healing will be lost and steps should be taken to protect it. He is also worried about the possible extinction of useful medicinal plants as civilization encroaches on remote areas. PMID- 7497403 TI - Training and recruitment of aboriginal public health workers--Phase II. PMID- 7497404 TI - Healthy elderly: coming of age. PMID- 7497405 TI - Frailty: a public health perspective. AB - Traditionally, frailty has been conceptualized as reflecting an inevitable decline in abilities usually associated with physical aspects of aging. More recent thinking now sees frailty as reflecting an interaction among individual factors and a range of environmental elements. These emerging models, however, continue to consider frailty as a condition that resides within the individual rather than a situation that exists for the individual. We present a new model that defines frailty as occurring when there is diminished ability to carry out the important practical and social activities of daily living. Factors related to the occurrence of frailty are considered, as are the theoretical and practice implications of viewing frailty as a social construction. PMID- 7497406 TI - The quality of life of seniors living in the community: a conceptualization with implications for public health practice. AB - Maintaining and improving the quality of life (QOL) of seniors in the community is an increasingly important goal of public health planning and programming. To provide authentic assessments of seniors' quality of life, a partnership of public health workers and university researchers, working closely with seniors in the community, developed the Quality of Life Profile: Seniors Version (QOLPSV). Administration of the QOLPSV to a wide range of North York, Ontario seniors (n = 205) participating in community-based healthful living programs provided evidence of the instrument's psychometric integrity. Additionally, results indicated specific areas of need among seniors, suggesting shifts in emphasis in program delivery. Additional public health applications of a quality of life perspective are outlined. PMID- 7497407 TI - Elderly clients' perceptions: individual health counselling and group sessions. AB - This descriptive, exploratory study examined elderly clients' perceptions of individualized health counselling and group wellness sessions that were offered by public health nurses in clinic settings. The sample included 36 elderly persons living independently in the community. The participants reported that individualized health counselling and group wellness sessions provided different kinds of benefits and services. Monitoring of health status, support, and health teaching were valued nursing activities within individualized health counselling. Participants said that group wellness sessions were an effective means of acquiring general knowledge about aging and lifestyle choices but this format could not totally meet all their health care needs. Regardless of clinic type, blood pressure monitoring was viewed as important. Recognition of elderly clients' perceptions is crucial in planning and implementing programs that are appropriate and acceptable for improving the quality of life for this age group. PMID- 7497408 TI - [Tuberculosis in aged persons residing in Quebec facilities: are screening and chemoprophylaxis indicated?]. AB - FOCUS: Different strategies for the prevention and control of tuberculosis in facilities providing long-term care (LTCF) to the elderly were proposed by the U.S. Public Health Service (USPHS) and the Quebec Ministry of Health and Social Services (MHSS). Controversies regarding the screening and chemoprophylaxis of tuberculosis in the elderly were reviewed, the epidemiology of tuberculosis in the United States and Quebec was compared and the number of cases expected annually was estimated from estimates available in the literature. METHOD: Fifty five scientific articles and official documents identified in the Medline database were analyzed. RESULTS: U.S. epidemiologic data support screening and chemoprophylaxis but in Quebec, the absence of reported increased risk in LTCF explains the reservations of the MHSS. According to our estimates, expected cases of tuberculosis might be underestimated. CONCLUSION: In Quebec, the best strategy to control tuberculosis is not yet obvious. A precise estimation of the number of undiagnosed cases in this environment would help clarify the dilemma. PMID- 7497410 TI - Fatal farm injuries in Ontario, 1984 through 1992. PMID- 7497409 TI - Tractor-related injuries in Ontario. PMID- 7497412 TI - Less common causes of accidental drownings in Alberta. PMID- 7497411 TI - Regional variations of northern health: the epidemic of fatal trauma in northeastern Ontario. AB - OBJECTIVE: To examine the causes of traumatic death in a northern region of Ontario. METHODS: Prevalence study of trauma deaths occurring within the region of Northeastern Ontario over the years 1989-1991; regional data were compared with provincial data. RESULTS: 1,027 patient records were identified over the study period (51.4 deaths per 100,000 population/year). Non-intentional trauma accounted for 70% of all trauma deaths in the region; suicide (25%; 12.8/100,000) and homicide (5%; 2.4/100,000) were less common. Motorized vehicle trauma accounted for most of the non-intentional traumatic death (39%; 20.4/100,000). Age-standardized mortality ratios were 67% above the provincial average for non intentional trauma, 71% above the provincial average for suicides, 55% above the provincial average for homicides, and 68% higher for all forms of traumatic death. CONCLUSIONS: Traumatic death is a major health problem in northern areas; reduction of these rates depends on the development of an effective injury prevention strategy. PMID- 7497413 TI - Mortality attributable to tobacco use in Canada and its regions, 1991. AB - Using the data from the 1991 General Social Survey, Canadian Mortality Database, the 1991 Annual Report of Fire Losses in Canada and previously published estimates of lung cancer deaths attributable to passive smoking, the Smoking Attributable Mortality, Morbidity and Economic Cost method was used to estimate national and regional smoking-attributable mortality (SAM) for 1991. The results indicate that 27,867 men and 13,541 women died as a result of smoking, including 171 children under the age of one. This total of 41,408 deaths represents an increase of 3,051 deaths since 1989, with women accounting for 2,721 of these increased deaths. The increase in female mortality is almost entirely due to adult diseases, equally divided between neoplasms (990), cardiovascular diseases (927) and respiratory diseases (821). PMID- 7497414 TI - [Is municipal regulation of tobacco use applicable to Laval, Quebec?]. PMID- 7497415 TI - Parental smoking and risk of otitis media in pre-school children. AB - This study was designed to estimate the role of passive smoking in the occurrence of repeated acute otitis media among pre-school children. The parents of 918 children, who were part of a birth cohort, were interviewed when the children were four years old. The frequency of episodes of otitis media was determined, as well as the age at which the first such episode occurred. Information about smoking in the home had been obtained when the child was two weeks old and again at 6, 12, 18 and 40 months of age. In all, 119 mothers (13%) smoked at least 20 cigarettes a day on average, 205 (22%) smoked less than 20, and 593 (65%) stated that they did not smoke at all. Children of mothers who smoked 20 or more cigarettes per day were at significantly increased risk of having four or more episodes of acute otitis media: relative risk (RR) = 1.8, 95% confidence interval (CI) = 1.1-3.0 and of having their first episode earlier in life RR = 1.3 (CI = 1.0-1.8), after adjustment. Moreover, the risk of recurrent (> or = four episodes) otitis media increases with the amount of cigarettes smoked. These findings clearly strengthen the case for persuading parents not to smoke in the presence of young children. PMID- 7497416 TI - The impending Canadian prostate cancer epidemic. AB - PURPOSE: To model and forecast prostate cancer incidence and mortality in Canada to the year 2016. METHODS: Bivariate multiplicative models of prostate cancer incidence and mortality for Canadian men aged 45 years or older, linear in time and Weibull in age, were fitted using weighted non-linear regression. RESULTS: The number of incident cases of prostate cancer is forecast to increase from 11,355, observed in 1990, to 26,900 by the year 2010 and to 35,200 by the year 2016. The number of deaths are estimated to climb from 3,424, observed in 1991, to an estimated 6,300 by the year 2010, and to 7,800 by the year 2016. CONCLUSIONS: The dramatic increase in prostate cancer rates with age, coupled with the expected large increase in the elderly Canadian male population and steadily increasing prostate cancer incidence rates will produce very large increases in the number of men who will have prostate cancer over the next 20 years. This has important implications for health care delivery in the future. PMID- 7497419 TI - Alternative therapies in veterinary medicine. PMID- 7497420 TI - Veterinary education and manpower concerns. PMID- 7497418 TI - The American Veterinary Chiropractic Association. PMID- 7497417 TI - Does play equipment conform to the Canadian standard? AB - In the summer of 1991, play equipment in 254 playgrounds located on the island of Montreal were inspected, using a checklist made up of items drawn from the Canadian standard for the safety of children's playspaces and equipment. The results of the study, covering 605 climbers, 522 swings and 181 slides, made it possible to identify the most and least respected aspect of safety. For example, one out of two pieces of play equipment was installed on a protective surface that did not conform to the Canadian standard; seven out of ten swings had seats made of non-impact-absorbing materials; and six out of ten pieces of equipment had head entrapment openings. Knowing the physical shortcomings of play equipment is an important step in reducing injuries sustained on it. However, to be effective, the prevention of injuries related to play equipment requires a concerted effort on the part of several partners. PMID- 7497422 TI - Morris Animal Foundation supports mountain gorillas. PMID- 7497421 TI - Harassment in the workplace. PMID- 7497423 TI - An ethicist's commentary on whether veterinarians should prescribe medications that are neither therapeutic nor prophylactic to increase productivity. PMID- 7497424 TI - Surgical management of preputial injuries in bulls: 51 cases (1986-1994). AB - The purpose of this study was to compare success rates for preputial surgery depending on the initial diagnosis, ability to extend the penis, use of sedation and local anesthesia versus general anesthesia for surgery, and surgical technique. Medical records of 51 bulls treated surgically for preputial injury were reviewed. The mean age of the bulls was 2.5 years ranging from 1 to 5 years. Bos taurus breeds (82.3%) were more often affected than Bos indicus breeds (17.7%). The most common breeds represented in this study were Angus (45.1%), Simmental (11.8%), and brangus (9.8%). The seasonal incidence of preputial injuries was higher during the period of May-July (52.9%) and November-February (33.3%). The overall success rate was 70%. Posthioplasty was more successful than circumcision (90% to 43%) (P < 0.05). The success rate for surgeries performed under inhalation anesthesia in the surgery suite was 100% compared with a success rate of 63% for those animals operated on with injectable and local anesthesia in a rotary chute (P < 0.05). The success rate was 88% if the penis could be extended before the surgery and 36% if extension was not achieved (P < 0.05). PMID- 7497425 TI - The effects of changing demographics on the distribution of marine anemia in farmed salmon in British Columbia. AB - The changing geographic distribution of marine anemia (plasmacytoid leukemia) was compared with the evolving demographics of the chinook farming industry in British Columbia to explore the hypothesis that the disease had spread throughout the province between 1987 and 1992. Through retrospective and prospective methods, it was shown that the apparent spread of the disease was likely an artifact, resulting from changes in the distribution of fish farms throughout the province and corresponding changes in the intensity of regional disease surveillance. When viewed over a 5-year period, there were no statistically significant differences in the prevalence of the disease amongst fish sampled from each of the major salmon farming regions of British Columbia. By increasing the intensity of surveillance for the disease in apparently negative regions or chinook farms, one could routinely find cases of the disease. The results suggest that marine anemia is an endemic problem for farmed chinook salmon in British Columbia and is not a spreading epidemic. PMID- 7497426 TI - Intestinal adenocarcinomas in two beluga whales (Delphinapterus leucas) from the estuary of the St. Lawrence River. PMID- 7497427 TI - Encephalitic nematodiasis in a Douglas squirrel and a rock dove ascribed to Baylisascaris procyonis. PMID- 7497429 TI - Mechanobullous disease in a Belgian foal in eastern Ontario. PMID- 7497430 TI - Mineralized paraprostatic cyst in a dog. PMID- 7497428 TI - Cerebellar cortical degeneration in a Labrador retriever. PMID- 7497431 TI - Prophylaxis: what is needed to do a good job? PMID- 7497433 TI - Critical thinking: how the experts do it. PMID- 7497432 TI - Insulin therapy for dogs and cats. AB - Management of diabetic dogs and cats requires a tremendous cooperative effort between the practitioner and the client. Consistency in the handling, availability, and formulations of the different insulins will improve client compliance. In addition to insulin therapy, successful management of the diabetic animal includes the client's perceptions of the animal's health, maintenance of the animal's body weight, consistency in water consumption, and monitoring serial blood glucose concentrations. Serial blood glucose determinations improve the practitioner's ability to identify and address problems associated with insulin therapy, and thereby delay or minimize the complications of long-term diabetes. PMID- 7497434 TI - Diseased fish submissions at Canadian pathology laboratories before 1910: historical review of the cases. PMID- 7497435 TI - Cytogenetics of malignant gliomas: I. The autosomes with reference to rearrangements. AB - Autosomal chromosome abnormalities are far from always detectable and, when detected, far from fully consistent in malignant gliomas. In 15 of 41 malignant gliomas, we found autosomal chromosome aberrations ranging from solitary trisomy to a wildly abnormal polyploid complement. The sequence of chromosome events appears to proceed from the normal to the near-diploid state (via structural and numerical changes) to near-tetraploidy (via polyploidization), and finally toward near-triploidy (via chromosome loss and additional rearrangements). Characteristic chromosome changes of trisomy 7 and monosomy 10 were repeatedly found, usually together in the same cell clones. In only one case was trisomy 7 an isolated change. We observed structural rearrangements of chromosomes 7 and 10 which may be of some use in mapping specific genes duplicated or deleted by the whole-chromosome changes of chromosomes 7 and 10. Nonrandom structural changes of other autosomes, including chromosomes 1, 5, and 11, fit with the model of malignant glioma as a process involving multiple genes. An unusual concentration of breakpoints in 12q13, juxtaposing it to at least five other regions, reflects the presence of genetic information in 12q13 important to the development of malignant gliomas. PMID- 7497436 TI - Cytogenetic analysis is non-informative for assessing the remission rate in chronic myeloid leukemia (CML) patients on interferon-alpha (IFN-alpha) therapy. AB - Cytogenetic analysis is considered pivotal for assessing the remission rate in CML patients on IFN therapy. On the basis of general agreement, at least 25 metaphases should be analyzed in each case. The main limitations to this approach are: 1) the small number of analyzable metaphases generally found in cytogenetic preparations from IFN-alpha-treated patients; and 2) the inability of this technique for scoring interphase cells. We compared the results of cytogenetic analysis and double-color FISH detection of bcr/abl genes fusion in 13 CML patients on IFN-alpha therapy (marrow sampling for cytogenetic and FISH analysis was carried out after 12 months in all patients and repeated after 18 months of IFN therapy in patients 4, 6, and 8). In five specimens, 20 to 25 cells were evaluable for cytogenetic examination, in another five no analyzable metaphases were scored, and in the remaining six samples two to 14 cells could be analyzed. With FISH detection at least 100 cells were easily scored in each specimen (mean number, 175). Comparing the results carried out with the two methods in different samples it emerged that cytogenetic analysis led to improper conclusions as regards the rate of Ph positivity, even in those patients where 20-25 metaphases were analyzed. Although many more cases have to be studied to establish the role of FISH analysis in Ph-positive patients, we are of the opinion that cytogenetic analysis is unfit for easily and accurately assessing the actual quality of remission in IFN-treated subjects. PMID- 7497437 TI - Partial triplication (7q) in a child with acute lymphoblastic leukemia demonstrated with conventional cytogenetics, PRINS, and chromosome painting. AB - In this report we describe a partial triplication (7q) and other structural aberrations found in a child with acute lymphoblastic leukemia (ALL), and we demonstrate the importance of PRimed IN Situ labeling (PRINS) and chromosome painting as a support to banding analysis for the clarification of complex structural chromosome rearrangements. Initially, the der(7) was interpreted as der(7)t(1;7;7). However, PRINS and chromosomes painting showed that der(7) consisted entirely of chromosome 7 material. Further, a derivative chromosome interpreted by banding analysis as a der(17)t(?1;17) was shown to be der(17)t(13;17) by the newly developed PRINS painting technique. PMID- 7497438 TI - Neurofibromatosis 2 gene in human colorectal cancer. AB - Colon cancers commonly have allelic losses of chromosome 22q, which suggests the presence of a tumor suppressor gene on 22q. The candidate tumor suppressor gene on 22q is the neurofibromatosis 2 (NF2) gene. Using single strand conformation polymorphism (SSCP) analysis, we screened 24 pairs of colorectal cancer and adjacent normal mucosa, as well as 10 colon cancer cell lines from non-NF2 patients, for mutations in the coding sequence of the NF2 gene. Two SSCP variants, one in exon 14 and another one in exon 16, were detected in two of the sporadic colorectal cancers, but not in adjacent normal mucosa samples. Sequencing of these variants in one tumor detected an A-to-G transition in bp 1459 of the NF2 cDNA, resulting in the change of Ile to Val at codon 487 of merlin, the NF2 protein product. The other tumor showed a 2-bp (CT) deletion in the intronic sequence of the alternatively spliced exon 16. These results suggest that the NF2 gene is probably involved in some colorectal tumors, but is not the critical chromosome 22q tumor suppressor gene involved in colon tumorigenesis. PMID- 7497439 TI - Cytogenetics of fine needle aspiration biopsies of sarcomas. AB - Chromosomal analysis has become an adjunct in the classification of soft tissue and bone sarcomas and may provide indications of tumor progression. Since it is not always possible or desirable to obtain fresh tumor tissue, the current study analyzed the feasibility to obtain karyotypes from fine needle aspiration biopsies (FNABs) and compared those with karyotypes obtained after short term tissue culture of the corresponding surgical specimens. In 6 of 12 cases an interpretable abnormal karyotype was obtained from the FNABs. These findings demonstrate that it is indeed feasible to obtain karyotypes of primary, metastatic or recurrent sarcomas after a minor procedure such as a FNAB. PMID- 7497440 TI - Translocation 11;14 in three children with acute lymphoblastic leukemia of T-cell origin. AB - Clinical, karyotypic, immunophenotypic, and molecular profiles of three TALL cases carrying a t(11;14) are discussed and compared with data in the literature. As previously reported, t(11;14)(p13;q11) was associated in one patient with a TALL profile of intermediate stage of maturation (CD7+, CD4+, CD8+). However, the same translocation was found to be present in another patient with a more immature, pro-TALL profile (CD7+, CD4-, CD8-). Both patients showed molecular rearrangements of the TCR beta chain gene. A third patient, with a very immature pro-TALL profile (CD34+, CD7+, CD4-, CD8-), carrying a t(11;14)(p15;q11), showed molecular rearrangements of the TCR beta and gamma chain genes, while the IgH chain genes were in germline configuration. Our data indicate that t(11;14) can also be present in TALLs of more immature stages of intrathymic development; the significant factor determining the clinical behavior of TALLs is apparently related more to cell differentiation than to the presence of this chromosome rearrangement. PMID- 7497442 TI - Cytogenetic evidence that a tumor suppressor gene in the long arm of chromosome 1 contributes to glioma growth. AB - In a patient with a rare subtype of glioma, pleomorphic xanthoastrocytoma, cytogenetic studies revealed that both homologues of chromosome 1 were involved in translocations at the same band 1q42 but with different partner chromosomes. In addition, 5 glioblastomas out of 25 gliomas karyotyped in our laboratory had lost at least one copy of band 1q42 through deletions, unbalanced rearrangements, or chromosome losses. Twenty-one gliomas that had lost at least one copy of chromosome band 1q42 were identified in the literature; all were astrocytic tumors and the majority were glioblastomas. It indicates a covert tumor suppressor gene in the region that is involved in astrocytic gliomas. PMID- 7497441 TI - Multiple chromosomal aberrations and 11p allelotyping in lung cancer cell lines. AB - Cytogenetic and molecular genetic studies have implicated many chromosomal aberrations in the pathogenesis of lung cancer. Deletions on 3p and 9p are presently the primary target for positional cloning of putative tumor suppressor genes. We have recently reported frequent loss of heterozygosity in three separate regions (HRAS, D11S12, D11S16) on 11p in freshly resected lung cancer specimens. Here we report cytogenetic and molecular genetic analyses of 26 permanently growing human lung cancer cell lines. Deletions indicating regions which may harbor potential tumor suppressor genes were found in 5/9 cell lines on 2p, 5/9 on 2q, 6/9 on 3p, 7/9 on 3q, 5/9 on 6q, 3/9 on 9p, 5/9 on 11p, and 6/9 on 13q. Reduction to hemizygosity or a statistically significant increase in the frequency of homozygosity on 11p was found for all markers investigated except for ST5 (D11S832E). Eight of twenty-six (31%) cell lines were hemizygous for D11S12 and 9/26 (35%) for D11S16. Seventeen of eighteen (94%) cell lines were homozygous for PTH (expected homozygosity, 53%), 15/15 (100%) for WT1 (expected homozygosity, 55%), and 16/18 (89%) for CAT (expected homozygosity, 50%). These results confirm the notion that 11p harbors several putative tumor suppressor genes which may become inactivated at different stages of tumor development and progression. They also provide a basis for selecting cell lines for genetic complementation specifically targeted at the regions described. PMID- 7497443 TI - Expression of common fragile sites in lymphocytes of Wilms tumor in patients, their parents, and siblings. AB - Fragile site expression in blood lymphocytes of 56 individuals, including 11 Wilms tumor patients, family members, and controls, was studied by 5 fluorodeoxyuridine induction with caffeine enhancement. Significantly elevated expression of fra(11)(p13) was observed in the patient group, compared with controls. One patient clinically diagnosed with the Wilms tumor, aniridia, and genitourinary and retardation (WAGR) syndrome revealed constitutional deletion of 11p13. Expression of fra(11)(p13) in this individual was observed only on the nondeleted homologue. Among the family members studied, only one sibling showed elevated expression of fra(11)(p13). PMID- 7497444 TI - Analysis of microsatellite repeats in pediatric brain tumors. AB - Tumorigenesis has been shown to proceed through a series of genetic alterations involving protooncogenes and tumor suppressor genes. However, investigation of genomic instability of microsatellites has disclosed a new mechanism for human carcinogenesis, which is involved not only in hereditary nonpolyposis colon cancer (HNPCC) but also in a number of other malignancies. To determine whether microsatellite instability is involved in pediatric brain tumors, we screened 15 such tumors using seven microsatellite marker loci on six chromosomes 4, 5, 9p, 9q, 11, 14, and 17. Using the polymerase chain reaction method, DNA samples from the tumors and from normal peripheral blood leukocytes from each patient were compared for the allelic pattern produced at each locus. Our preliminary results indicate loss of heterozygosity at the fatty acid binding protein (FABP) locus, located on chromosomal arm 4q28-q31, the only trinucleotide repeat in the panel of markers used, for 3 of 15 cases, suggesting the presence of previously unidentified sequences relevant to brain tumorigenesis at or in the vicinity of this locus. We did not observe any microsatellite instability in these samples, indicating that the mechanisms operating in HNPCC are not active in this subset of pediatric brain tumors. PMID- 7497445 TI - Correlation of cytogenetic results with immunophenotype, genotype, clinical features, and ras mutation in acute myeloid leukemia. A study of 235 Chinese patients in Taiwan. AB - Of 235 consecutive patients with de novo acute myeloid leukemia (AML), clonal chromosomal abnormalities were detected in 151 (64%) of them. Twenty-four of the 71 patients with M2 AML had t(8;21), 35 of the 36 M3 patients had t(15;17), and 11 of the 45 M4 leukemia disclosed inv(16). Six of the eight patients with 11q23 abnormality had M4 or M5 subtype of leukemia. The incidence of t(15;17) and t(8;21) was higher in our patients than in patients from most Western countries. Immunophenotyping was performed on 197 patients. Patients with t(15;17) were associated with negativity to HLA-DR, CD11b, and CD34. Patients with t(8;21) expressed CD13 and CD33 less frequently than other patients, but all showed CD15 positivity. Coexpression of lymphoid-associated antigens on the leukemic blasts was detected in 52 patients (26%), including all 7 patients with t(9;22), 3 of the 8 patients with t/del(11)(q23), 2 of the 25 patients with t(15;17), and 2 of the 22 patients with t(8;21). Seven (35%) of the 20 patients coexpressing lymphoid markers showed immunoglobulin heavy chain or T-cell receptor beta-chain gene rearrangements, while only 2 (4%) of the 53 patients without lymphoid antigen expression did so. Patients with inv(16), t(8;21), and t(15;17) had a better prognosis than other patients. Of all surface antigens tested, only CD15, CD11b, and HLA-DR were of prognostic value: CD15 with a higher complete remission (CR) rate and CD11b or HLA-DR with a shorter CR duration. N-ras mutations were detected in 7 (18%) of the 40 patients in the study, including two of the three patients with inv(16). This study demonstrated differences in clinical features, immunophenotypes, and genotypes among different cytogenetic subgroups. PMID- 7497446 TI - Overexpression of the PRAD1 oncogene in a patient with prolymphocytic leukemia with t(11;14)(q13;q32). AB - Prolymphocytic leukemia (PLL) was diagnosed by morphologic and immunophenotypical studies in a 72-year-old Japanese man. Massive splenomegaly was present but lymphadenopathy was minimal in this case. Chromosomal analysis of peripheral mononuclear cells showed t(11;14)(q13;q32) in all metaphases examined, except for one normal karyotype. Northern blot analysis of RNA prepared from leukemic cells obtained from the patient revealed overexpression of the PRAD1/cyclin D1 proto oncogene, which has not been described previously in patients with PLL. PMID- 7497447 TI - Chromosome 7 in glioblastoma tissue. Parenchymal vs. endothelial cells. AB - Strong endothelial proliferation is a prominent feature of glioblastomas and sometimes these proliferated areas transform into a malignant component of glioblastoma, resulting in gliosarcomas. It has not been established whether the proliferated endothelial areas are cytogenetically abnormal. To clarify this question, the most common cytogenetic aberration, gain of chromosome 7, was chosen and in situ hybridization was performed on paraffin-embedded tissue sections of three glioblastomas. The purpose was to compare the parenchymal tumor cells and the endothelial cells. The results showed trisomy 7 in only a small amount of endothelial cells (5-8%), whereas 23-38% of parenchymal tumor cells displayed trisomy 7. PMID- 7497449 TI - A hamartoma of the breast with an aberration of 12q mapped to the MAR region by fluorescence in situ hybridization. AB - Cytogenetic studies of a breast adenolipoma (hamartoma) of a 58-year-old patient revealed a karyotype 46,XX,add(4)(?),add(6)(q?),der(7)t(7;12)(q11.1 or q11.2;q11 or q12),der(12). To our knowledge, this is the second report of an aberration involving 12q12-15 in a hamartoma of the breast. By FISH studies, we found this chromosome 12 translocation breakpoint to be mapping within the MAR (Multiple Aberration Region). MAR is known to be a major cluster region of chromosome 12 breakpoints of benign solid tumors such as uterine leiomyoma, lipoma, and pleomorphic salivary gland adenomas, therefore raising the possibility that the same gene is involved in hamartoma of the breast as in these three benign solid tumors. PMID- 7497448 TI - Detection of chromosome 18 rearrangement in synovial sarcoma by fluorescence in situ hybridization. AB - Fluorescence in situ hybridization (FISH) evaluations of chromosome 18 were performed in synovial sarcoma, hemangiopericytoma, and mesothelioma. Each case was evaluated with centromeric and whole chromosome paint probes. The synovial sarcomas had t(X;18) cytogenetically, but the FISH evaluator was blinded to the cytogenetic results and to the histopathologic diagnosis. The FISH analyses were consistent with chromosome 18 translocation in 6 of 7 synovial sarcomas, 0 of 3 hemangiopericytomas, and 0 of 1 mesothelioma. These findings support the use of FISH in the diagnosis of synovial sarcoma. PMID- 7497450 TI - Endometrial stromal sarcoma with clonal chromosomal aberrations and mixed phenotype. AB - We report a case of a moderate-grade endometrial stromal sarcoma with the following chromosomal complement based on the evaluation of 43 metaphases: 47,XX,der(3)t(3;6)(q29;p21.1),der(6) t(3;6)(q21;q27), + 19. Immunohistochemically, the paraffin-embedded tumor tissue displayed positive vimentin reactivity and lack of cytokeratin expression, indicating a mesenchymal origin. Interestingly, the cultivated tumor cells revealed a co-expression of vimentin and different subtypes of cytokeratin. Therefore, the cytogenetically monoclonal tumor cells which showed co-expression of epithelial and mesenchymal phenotypes suggest that the endometrial stromal sarcoma can be interpreted as a monophasic variant of the malignant mixed Mullerian tumor. PMID- 7497451 TI - Chromosomal abnormalities in tumor and lymphocyte cultures from patients with pituitary adenomas. PMID- 7497452 TI - Cytogenetics of malignant gliomas. II. The sex chromosomes with reference to X isodisomy and the role of numerical X/Y changes. AB - Sex chromosomal monosomy with total loss of an X or Y is frequently observed in malignant gliomas. Beyond that, not much is known about the behavior of the sex chromosomes in these tumors. We noted loss of the X from 3 of 13 gliomas from women (23%) compared to loss of the Y from 16 of 28 gliomas from men (57%). There were two structural rearrangements of the Y (an inversion and a translocation with chromosome 4). Most unexpectedly, clones with sex chromosome reversal were encountered in 3 cases. These XX clones in gliomas from men are perforce the consequence of Y loss coupled with X isodisomy, a nonrandom sequence of sex chromosome changes. We examined the company kept by numerical X and Y changes in clones and found that clones with numerical sex chromosome changes had fewer autosomal abnormalities, reflecting a distinct tendency to clonal separation of sex chromosome from autosomal abnormalities. We conclude that the sex chromosome changes are not a necessary part of the neoplastic process in malignant gliomas but that they must be of biologic significance to the brain since they are highly nonrandom in frequency, type, and sequence in brain cells. PMID- 7497453 TI - High chromosome numbers of testicular germ cell tumors. An update. PMID- 7497454 TI - Induction of CYP isoenzymes in various organs of rats by 3-methylcholanthrene or beta-naphthoflavone. AB - Induction of cytochrome P450 (CYP) isoenzymes in various organs of rats treated with 3-methylcholanthrene (3-MC) and beta-naphthoflavone (BNF) was immunohistochemically and biochemically investigated. Fifteen male F344 rats were divided into three equal groups. Group 1 was untreated as a control. On days 3, 4, 13 and 14, group 2 animals received 3-MC (20 mg/kg body wt i.p.) dissolved in corn oil while group 3 animals were given BNF (50 mg/kg body wt i.p.) dissolved in corn oil, at days 1, 2, 3, 4, 11, 12, 13 and 14. On days 4 and 14, two or three animals in each group were sacrificed, 15 h after administration of the last test compound. Induction of CYP 1A1, 2C11, 2D1, 2E1, 3A2 and 4A1 in various organs was immunohistochemically examined and the levels of CYP 1A1 protein were measured by Western blotting. 3-MC and BNF induced CYP isoenzymes not only in the liver, but also in the small intestine, large intestine, prostate and seminal vesicles. The results indicate that xenobiotic metabolism can occur in various organs. PMID- 7497455 TI - Expression of immunoreactive human hepatocyte growth factor in human esophageal squamous cell carcinomas. AB - Hepatocyte growth factor (HGF) is a potent mitogen for epithelial cells that promotes cell motility and invasiveness. In this study, we report that the human esophageal squamous cell carcinoma (SCC) shows a significant elevation of HGF concentration (600 +/- 416 ng/100 mg protein), compared to normal mucosa (80 +/- 183 ng/100 mg protein) (P < 0.01). An association could be established between levels of HGF and decreasing differentiation of 37 SCCs. The 2-year crude survival rates were 51.1% and 68.4% at high and low HGF concentrations, respectively. The results indicate that HGF is significantly increased in human esophageal SCCs, especially of poorly differentiated type. HGF might thus be useful as a biological biomarker for characterization of human esophageal SCCs. PMID- 7497456 TI - Synergistic induction of apoptosis in breast cancer cells by tamoxifen and calmodulin inhibitors. AB - Breast cancer cells are relatively resistant to the induction of apoptosis (AP) and drug regimens which readily activate apoptotic death, may enhance the antitumor effect. Rapid and intensive induction of apoptosis was observed in estrogen receptor positive and negative breast cancer cell cultures treated with tamoxifen (TMX) combined with the calmodulin antagonists trifluoperazine (TFP) or W7. TMX (1-5 microM) alone or calmodulin antagonists alone did not induce apoptosis. Importantly, intensive apoptosis was also induced by TMX and TFP in the cells obtained from primary human breast carcinomas. Inhibition of the Ca2+ calmodulin signaling pathway is an effective way to activate apoptotic death in epithelial cells. Combination of TMX with non-toxic calmodulin inhibitors may increase the preventive and therapeutic effects of TMX. PMID- 7497457 TI - Ipomeanol analogs as chemopreventive agents: effect on the in vitro metabolism of 4-(methylnitrosamino)-1-(3-pyridyl)-1-butanone (NNK). AB - 4-(Methylnitrosamino)-1-(3-pyridyl)-1-butanone (NNK) is a tobacco-specific, powerful, organospecific lung carcinogen. 4-Ipomeanol (IPO) is an investigational chemotherapeutic drug with specific toxicity towards the lung. We hypothesized that non-toxic analogs of IPO could be competitive inhibitors of the metabolic activation of NNK. We had shown previously that 4-hydroxy-1-phenyl-1-pentanone (HPP) and 7-hydroxy-1-phenyl-1-octanone (4-HPO) are effectively inhibiting the lung tumor activity of NNK in A/J mice. In these extended studies we have synthesized 11 new analogs of HPP and tested them for their in vitro activities as inhibitors of the metabolism of NNK. The present study demonstrated that the lipophilicity in the molecule is playing an important role for the inhibition of NNK metabolism with pulmonary and hepatic microsomes of A/J mice. PMID- 7497459 TI - Modulation of expression of genes encoding nuclear proteins following exposure to JANUS neutrons or gamma-rays. AB - Previous work has shown that exposure of cells to ionizing radiations causes modulation of a variety of genes, including those encoding c-fos, interleukin-1, tumor necrosis factor, cytoskeletal elements, and many more. The experiments reported herein were designed to examine the effects of either JANUS neutron or gamma-ray exposure on expression of genes encoding nucleus-associated proteins (H4-histone, c-jun, c-myc, Rb, and p53). Cycling Syrian hamster embryo cells were irradiated with varying doses and dose rates of either JANUS fission-spectrum neutrons or gamma-rays; after incubation of the cell cultures for 1 h following radiation exposure, mRNA was harvested and analyzed by Northern blot. Results revealed induction of transcripts for c-jun, H4-histone, and (to a lesser extent) Rb following gamma-ray but not following neutron exposure. Interestingly, expression of c-myc was repressed following gamma-ray but not following neutron exposure. Radiations at different doses and dose rates were compared for each of the genes studied. PMID- 7497458 TI - Loss of nm23 and Alu DNA in human lung cancer propagated in nude mice. AB - Freshly resected human non-small cell lung cancer (NSCLC) has been successfully transplanted and propagated subcutaneously in nude mice (Cancer Letters 61 (1991) 53-60). We used this model to study the changes of the human metastasis suppressor genes, nm23-H1 and nm23-H2, through the process of propagation and metastasis of human NSCLC. Using a non-radioisotopic Southern analysis, the nm23 H1 and nm23-H2 genes were detected without evidence of deletion in the early generations of the tumor grafts. These genes, however, were absent from the tumor grafts sampled past 4 generations of propagation and from all the propagated metastases originated from the subcutaneous grafts. Further restriction analysis revealed that only mouse DNA, but no human Alu DNA, was present in the tumor specimens which lacked the human nm23 genes. Thus, there is a loss of human DNA but a gain of mouse DNA in the propagated tumors originated from the transplanted human NSCLC. The mechanisms of loss of human DNA in these propagated tumors in nude mice have yet to be delineated. PMID- 7497460 TI - Cytotoxicity, cellular accumulation and DNA binding of oxaliplatin isomers. AB - Oxaliplatin (trans-l-1,2-diaminocyclohexane oxalato Pt(II); 1R,2R-dach, l-OHP), its trans-d isomer (1S,2S-dach) and cis-dach (1R,2S-dach) isomers were compared in in vitro testing against human ovarian carcinoma cell lines A2780, A2780/CP (cisplatin resistant), A2780/l-OHP (oxaliplatin resistant), colon carcinoma cell line HT-29, and murine leukemia cell lines L1210, L1210/CP (cisplatin resistant), and L1210/dach (tetraplatin resistant). The relative molar potency of the three complexes in all the cell lines except A2780/l-OHP and L1210/dach are trans-l > trans-d > cis-dach; in A2780/l-OHP they are trans-l = trans-d > cis-dach; in L1210/dach trans-l > trans-d = cis-dach. The A2780/l-OHP selected for trans-l resistance is 3.6-fold resistant to oxaliplatin, showed no resistance to trans-d isomer and is 6-fold resistant to cis-dach. However, L1210/dach which is selected for carboxyphthalato 1,2-dach (trans-dl) platinum(II) is 140-fold resistant to oxaliplatin, 73-fold resistant to trans-d, and 41-fold resistant to cis-OHP. The accumulation and DNA binding of platinum following a 2-h treatment of A2780 cells with each of the isomers (60 microM) is in the order of trans-l > cis-dach > trans-d which corresponded to the cytotoxicity of trans-l, but not the others. The data suggest that other processes, such as differential formation of specific adducts and/or repair may be involved. Of the three isomers l-OHP is the superior and its accumulation and DNA binding are consistent with its cytotoxicity. PMID- 7497461 TI - Incidence of pancreatic and testicular tumors in rats treated with ciprofibrate, a peroxisome proliferator. AB - It is hypothesized that hepatic tumors in rats induced by peroxisome proliferators is dependent on peroxisome proliferative effect of these compounds and the resulting oxidative stress. However, it is argued that since these compounds also induce tumors in pancreas and testes, the two organs in which there is no proliferation of peroxisomes, the carcinogenic effect is unlikely to be related to oxidative stress. To clarify this controversy we have systematically analyzed the incidence of pancreatic acinar cell foci and nodules, and testicular Leydig cell tumors in ciprofibrate treated and control rats. In animals treated with 0.025% ciprofibrate for 22 months the incidence of Leydig cell tumors and acinar cell lesions was 100% and 66%, respectively. In age matched controls the incidence of testicular and pancreatic lesions was 93% and 66%, respectively. These findings clearly demonstrate that the Leydig cell tumors and pancreatic lesions develop spontaneously and are not induced by ciprofibrate. PMID- 7497462 TI - Introduction of murine Il-4 gene into B16(F10) melanoma tumors by direct gene transfer with DNA-liposome complexes. AB - Il-4 is a highly pleiotropic cytokine which induces cytotoxic activity when present at the tumor site. Death of tumor cells probably depends on the appearance of an inflammatory infiltrate composed of eosinophils and macrophages. Regression of established tumor masses has been readily observed upon direct intratumor implantation of cells which constitutively produce high amounts of Il 4. We now report a similar potent anti-tumor effect achieved via direct gene transfer, i.e. by injecting Il-4 DNA complexed with cationic liposomes into B16(F10) melanoma tumor in vivo. PMID- 7497463 TI - Cytokine induction by 41.8 degrees C whole body hyperthermia. AB - The potential for 41.8 degrees C whole body hyperthermia (WBH) to enhance ionizing irradiation and cytotoxic chemotherapy without a commensurate increase in normal tissue toxicity is currently receiving renewed clinical interest. Additionally, WBH may have other biological sequela which may be clinically exploited. In this paper, data are summarized revealing the ability of WBH to induce elevated plasma levels of granulocyte-colony stimulating factor (G-CSF), interleukin-1 beta (IL-1 beta), interleukin-6 (IL-6), interleukin-8 (IL-8), interleukin-10 (IL-10), and tumor necrosis factor-alpha (TNF-alpha) within hours of WBH. Data regarding TNF-alpha shows induction in only a proportion of patients. No induction of C-reactive protein (CRP) or the following cytokines was observed: granulocyte macrophage-colony stimulating factor (GM-CSF), interferon gamma (IFN-gamma), interleukin-1 alpha (IL-1 alpha), interleukin-2 (IL-2), interleukin-4 (IL-4), interleukin-7 (IL-7), interleukin-11 (IL-11), interleukin 12 (IL-12), macrophage-colony stimulating factor (M-CSF), and macrophage inflammatory protein-1 alpha (MIP-1 alpha). Data regarding interleukin-3 (IL-3) and transforming growth factor-beta 1 (TGF-beta 1) were variable and inconclusive. The implications of these results to past and future clinical trials are discussed. PMID- 7497464 TI - Peritoneal exfoliative cytology and Ki-ras mutational analysis in patients with pancreatic adenocarcinoma. AB - This is the first description of the detection of pancreatic adenocarcinoma peritoneal metastasis by established radiolabeled polymerase chain reaction (PCR) based Ki-ras mutational analysis. The present study evaluates both routine cytology and Ki-ras mutational analysis in the detection of peritoneal micrometastases in 24 subjects with pancreatic adenocarcinoma compared to seven control cases of chronic pancreatitis and seven control cases of cholecystitis. Locoregional extension, vascular invasion, and distal metastases were confirmed in 21/24 (88%) of the subjects with pancreatic adenocarcinoma by compute tomography, angiography, endosonography, or laparoscopy. The most common site of histologically confirmed extrapancreatic involvement was the vasculature (29%), followed by the liver (25%), duodenum (17%), peritoneum (17%), and lymph nodes (12%). Peritoneal lavage cytology was positive in 3/24 (12%) cases of pancreatic carcinoma while Ki-ras codon 12 mutational analysis was positive in 2/24 (8%). Two histologically confirmed cases of peritoneal metastases were not detected by either methodology, while peritoneal lavage cytology detected malignant cells in one case with histologically confirmed lymph node metastasis. PMID- 7497465 TI - Studies on clastogenic and carcinogenic potency of tetramethyl thiuram disulphide. AB - Tetramethyl thiuram disulphide (Thiram, TMTD), a mutagen, is analysed for its clastogenic property by using micronuclei induction studies in the bone marrow of erythrocytes of male Swiss albino mice. Three concentrations of TMTD, i.e. 100, 150 and 200 mg/kg body wt did not significantly increase the micronuclei in polychromatic erythrocytes or the ratio of poly-to normochromatic erythrocytes. The tumour initiating (10 mg and 1 mg each in 0.2 ml DMSO) and promoting (1 mg in 0.2 ml DMSO) potency of TMTD were tested in female Swiss mice by two stage skin carcinogenic model. There was no significant increase in tumours in the TMTD treated groups, either as initiator or promoter. The results indicated nonclastogenicity and non-carcinogenicity of TMTD under these experimental conditions. PMID- 7497466 TI - Membrane and cytosolic protein phosphorylation patterns in the early stages of DEN-induced hepatocarcinogenesis in rats fed a high or low protein diet. AB - The membrane and cytosolic protein phosphorylation patterns in the early stages of diethylnitrosamine-induced rat liver carcinogenesis, promoted by 2 acetylaminofluorene in the diet plus partial hepatectomy (DEN-AAF-PH), were analyzed by two-dimensional gel electrophoresis in animals fed a low protein (5% casein) diet, or the original high protein (24% casein) diet, in order to modulate the development of GST-P-positive preneoplastic lesions. Compared with untreated controls, membrane and cytosolic protein phosphorylation patterns changed only slightly in low protein-fed rats 7 days post-hepatectomy, with no appearance of enzyme-altered hyperplastic foci in the liver sections. By contrast, high protein-fed animals demonstrated GST-P-positive preneoplastic lesions 7 days post-hepatectomy and several acidic and more basic high M(r) phosphorylated membrane (between 97 and 116 kDa) as well as cytosolic (between 97 and 200 kDa) proteins could be detected. In the presence of enzyme-altered hepatocytes in the liver sections, low protein-fed rats demonstrated at 60 days post-hepatectomy cytosolic protein phosphorylation patterns remarkably similar to those shown by 24% casein-fed animals at 7 days post-hepatectomy, suggesting close correlation between protein phosphorylation patterns and development of preneoplastic lesions during the early stages of DEN-AAF-PH liver carcinogenesis. This may arise by a constitutive activation of one or more signal transduction pathways, possibly involving protein kinase C, during liver tumour promotion. PMID- 7497468 TI - Improved genomic/nuclear DNA extraction for 8-hydroxydeoxyguanosine analysis of small amounts of rat liver tissue. AB - Using two different commercially available extraction kits, genomic/nuclear DNA could be recovered from rat liver samples as small as 10 mg. Background 8 hydroxydeoxyguanosine levels in such DNA were low and stable at 0.26-0.30 +/- 0.01-0.03/10(5) guanine residues. The minimum tissue wet weight required for the accurate 8-hydroxydeoxyguanosine analysis was 25 mg. The present results indicate that routine and reliable assessment of the involvement of oxidative DNA damage in the development of various diseases, including cancer, is feasible using a variety of tissue sources. PMID- 7497467 TI - Overexpressions of c-fos/jun mRNA and their oncoproteins (Fos/Jun) in the mouse uterus treated with three natural estrogens. AB - To further understand hormonal carcinogenesis of natural estrogens (estrone, 17 beta-estradiol (E2) and estriol), we determined the expressions of c-fos/jun mRNA, and their oncoproteins (Fos/Jun) with intracellular localization in the uterus of ovarectomized mice treated with these estrogens. Mid-term chronic, as well as short-term assays were examined. Of three estrogens examined, mid-term chronic E2-treatment significantly increased the expression of c-fos/jun mRNA, and their oncoproteins (Fos/Jun). These were most prominently expressed in glandular cells of E2-treated mouse endometrium. Therefore, mid-term chronic E2 treatment might partially induce glandular cell transformation of uterine endometrium via overexpression of Fos/Jun. PMID- 7497469 TI - Homozygous deletions of p16INK4A/MTS1 and p15INK4B/MTS2 genes in glioma cells and primary glioma tissues. AB - The p16INK4A/MTS1 (p16) and p15INK4B/MTS2 (p15) genes map to 9p21 where genetic alterations have been frequently reported in various human tumors. Using the polymerase chain reaction (PCR), we investigated the loss of these genes on primary glioma samples and cultured glioma cells. All or any of three exons of the p16 gene were homozygously delted in 11 (35.5%) of 31 glioblastomas, none of 9 anaplastic astrocytomas and 5 astrocytomas, and in all 6 human glioma cell lines. Exon 2 of the p15 gene was homozygously deleted in 4 (12.9%) of 31 glioblastomas, but not in lower grade gliomas. It was homozygously deleted in 5 (83.3%) of 6 glioma cell lines. In 12 short-term cultures of cells derived from primary glioma samples, 5 (41.7%) and 2 (16.7%) glioblastoma-derived cells had homozygous deletion of all or any of the three exons of the p16 gene and exon 2 of the p15 gene, respectively. The deletion pattern of these genes in cultured cells was completely consistent with that seen in the primary tumors. Furthermore, two long-term cultures retained both genes that were identical to those in the original tumor tissues. Our results indicate that loss of the p16 and p15 genes may be involved in tumor progression in human gliomas, especially in the development of glioblastoma, that this loss may give growth advantage to the cells in culture, and that it is not the result of culture artifacts. PMID- 7497470 TI - Accumulation of benzoporphyrin derivative in malignant and host cell populations of the murine RIF tumor. AB - The accumulation of benzoporphyrin derivative (BPD) in malignant and host cell populations of the mouse RIF tumor was studied by flow cytometry analysis of cells dissociated from the tumor tissue. The highest cellular BPD levels were found in a subpopulation of tumor associated macrophages (TAMs) characterized by a highly elevated expression of interleukin 2 receptor (F4/80+CD25++), but this selectivity was less pronounced than with photofrin. The BPD levels in tumor associated immune cells other than TAMs were lower than in malignant cells. No significant difference in cellular distribution of BPD was observed with the photosensitizer delivered in liposomes compared to the aqueous solution. PMID- 7497471 TI - Inhibition of metabolism of 4-(methylnitrosamino)-1-(3-pyridyl)-1-butanone by dietary benzaldehydes. AB - As part of a routine screening assay, benzaldehyde was found to inhibit 4 (methylnitrosamino)-1-(3-pyridyl)-1-butanone (NNK) metabolism. Consequently, the effects of benzaldehyde and several structurally related compounds on NNK metabolism were examined in murine hepatic and pulmonary microsomes. All test compounds inhibited formation of the metabolites 4-oxo-4-(3-pyridyl)butyric acid (OPBA), 4-hydroxy-1-(3-pyridyl)-1-butanone (HPB), and 4-(methylnitrosamino)-1-(3 pyridyl)-1-butanol (NNAL) in hepatic microsomes and inhibited formation of 4 (methylnitrosamino)-1-(3-pyridyl-N-oxide)-1-butanone (NNK N-oxide), HPB, and NNAL in pulmonary microsomes. m-Anisaldehyde was the most potent inhibitor, and p hydroxybenzaldehyde and syringaldehyde were less potent than benzaldehyde and vanillin in inhibiting the formation of OPBA and HPB, NNK metabolites that reflect metabolic activation (alpha-hydroxylation). Vanillin was essentially as potent as benzaldehyde. The mechanism of inhibition exhibited by these compounds appears to be competitive in nature. The ability of these compounds to inhibit NNK activation suggests that these compounds may be effective blocking agents (anti-initiating agents) for NNK lung tumorigenesis. PMID- 7497472 TI - Gemfibrozil-induced peroxisome proliferation and hepatomegaly in male F344 rats. AB - Gemfibrozil is a widely used hypolipidemic drug in humans that causes peroxisome proliferation and hepatocarcinogenesis in rodents. The induction of hepatomegaly and hepatic peroxisome proliferation (measured as peroxisomal acyl CoA oxidase activity), was determined and compared to another peroxisome proliferator, WY 14,643 (0.1% in the diet) in male F344 rats. In a 21-day study, dietary no observable-effect and lowest-observable-effect levels of gemfibrozil for both hepatomegaly and peroxisome proliferation were 0.002% and 0.005%, respectively. In a 42-day study, dietary concentrations of 0.9-2.0% gemfibrozil induced a similar magnitude of hepatomegaly to WY-14,643 (2.3-fold) but a higher level of peroxisome proliferation (16-18-fold) than the maximum induction for WY-14,643 (13-fold). The plateau in magnitude of gemfibrozil-induced peroxisome proliferation across the 0.9-2.0% dietary concentrations was associated with a plateau in serum concentration of gemfibrozil (approximately 20 micrograms/ml), similar to concentrations reported in human subjects receiving oral gemfibrozil. These results indicate that maximal induction of peroxisome proliferation by gemfibrozil can exceed that of a more potent compound such as WY-14,643, and further suggest that maximal induction of peroxisome proliferation can be limited by steady-state serum concentrations. Moreover, the reported lack of hepatic responses to gemfibrozil in humans is unlikely to be the result of inefficacy or unavailability of this drug, compared to other peroxisome proliferators, in rodents. PMID- 7497473 TI - Sequencing of xyloglucan oligosaccharides by partial Driselase digestion: the preparation and quantitative and qualitative analysis of two new tetrasaccharides. PMID- 7497475 TI - Synthetic and structural studies of alpha-sialyl-(2-->6) and alpha-sialyl-(2-->3) 1-deoxynojirimycin derivatives potentially useful for biomedical applications. AB - Suitably protected derivatives of 1-deoxynojirimycin (1,5-dideoxy-1,5-imino-D glucitol, DNJ) and its D-galacto analog were coupled with 2-thioglycosides of N acetylneuraminic acid. The resulting disaccharides were converted into a variety of alpha-sialyl-(2-->6)-and alpha-sialyl-(2-->3)-DNJ derivatives, including the cyclic lactams 6-O-(5-acetamido-3,5-dideoxy-D-glycero-alpha-D-galacto-2 nonulopyrano sylono- 1',5-lactam)-1,5-dideoxy-1,5-imino-D-glucitol and -D galactitol. The structural features of the synthetic compounds were investigated by ion-spray mass and 1H NMR spectrometry. The 1C4 conformation of N-tert butoxycarbonyl-DNJ, a synthetic intermediate having the gluco configuration, was confirmed by X-ray crystallography. PMID- 7497474 TI - Novel aminocyclitol antibiotics derived from natural carbohydrates. PMID- 7497476 TI - The chemical synthesis of Leishmania donovani phosphoglycan via polycondensation of a glycobiosyl hydrogenphosphonate monomer. AB - A polycondensation of 2,3,6-tri-O-benzoyl-4-O-(2,3,4-tri-O-benzoyl-beta-D galactopyranosyl)-al pha-D-mannopyranosyl hydrogenphosphonate in the presence of trimethylacetyl chloride has been used to synthesize a linear poly[beta-D galactopyranosyl-(1-->4)-alpha-D-mannopyranosyl phosphate] representing the phosphoglycan part of the lipophosphoglycan from Leishmania donovani. PMID- 7497477 TI - The role of deoxyhexonic acids in the hydrothermal decarboxylation of carbohydrates. AB - Hydrothermolysis of D-glucose, cellobiose, and beta-cyclodextrin leads to the formation of small amounts of 3-deoxyhexonic acids. These acids are known to be produced by the alkaline degradation of carbohydrates. The formation under neutral hydrothermal conditions of the 3-deoxyhexonic acids provides evidence for the formation of 3-deoxy-D-erythro-hex-2-ulose, a compound that has been reported to play a role in both alkaline and acidic conversion of carbohydrates. Hydrothermolysis of 2- and 3-deoxy-D-arabino-hexonic acid does not lead to significant decarboxylation, and therefore these compounds cannot be considered to play a major role in the initial hydrothermal decarboxylation of biomass. PMID- 7497478 TI - Structural studies of the saccharide part of the cell envelope lipooligosaccharide from Haemophilus influenzae strain galEgalK. AB - The structure of the saccharide part of the lipooligosaccharide from Haemophilus influenzae strain galEgalK has been investigated. On treatment of the lipooligosaccharide with acid under mild conditions, followed by reduction with sodium borohydride and gel permeation chromatography, a main fraction was obtained which was studied by methylation analysis, NMR spectroscopy, and FABMS. The material was heterogeneous and contained two major compounds, A and B, and one minor, C. [formula: see text] In the structure, PEA is phosphoethanolamine, and L-D-Hep is L-glycero-D-manno-heptose. Kdo exists in reduced anhydro forms. The carbohydrate backbone is the same as that proposed for the saccharide part of the major component from H. influenzae type b strain A2 [N.J. Phillips, M. A. Apicella, J. M. Griffiss, and B. W. Gibson, Biochemistry, 32 (1993) 2003-2012]. PMID- 7497479 TI - Structure of the O6 antigen of Stenotrophomonas (Xanthomonas or Pseudomonas) maltophilia. AB - A polysaccharide containing D-xylose, L-rhamnose, and N-acetyl-D-glucosamine was released on mild acid hydrolysis of the lipopolysaccharide extracted from defatted cell walls of Stenotrophomonas (Xanthomonas or Pseudomonas) maltophilia strain 557, the reference strain for serotype O6. By means of NMR spectroscopy and chemical degradations, the repeating unit of the polymer was identified as a branched trisaccharide with the structure shown. [formula: see text] PMID- 7497480 TI - Characterization of a lipopolysaccharide O antigen containing two different trisaccharide repeating units from Burkholderia cepacia serotype E (O2). AB - The O antigen of the lipopolysaccharide of Burkholderia cepacia serotype E (O2) was shown by a combination of methylation analysis, partial hydrolysis, NMR, and mass spectrometric methods to be a high molecular weight polysaccharide composed of two different trisaccharide repeating units in the ratio 2:1. The major trisaccharide component is composed of two alpha-D-mannopyranosyl and one beta-D galactopyranosyl residues with the structure, [-->2)-alpha-D-Man p-(1-->2)-alpha D-Man p-(1-->4)-beta-D-Gal p-(1-->]n The minor trisaccharide component is a D mannan composed of two alpha- and one beta-D-mannopyranosyl residues with the structure, [-->2)-alpha-D-Man p-(1-->2)-alpha-D-Man p-(1-->3)-beta-D-Man p-(1- >]n PMID- 7497481 TI - Isolation and partial characterization of feruloylated oligosaccharides from maize bran. AB - Maize bran contains phenolic acids [approximately 4% dry matter; mainly ferulic acid (Fe) and also diferulic acid], heteroxylans (approximately 50%), and cellulose (approximately 20%), but is devoid of lignin. Treatment of maize pericarp with 0.05 M trifluoroacetic acid at 100 degrees C for 2 h released approximately 90% of the heteroxylans and approximately 90% of the ferulic acid and its esters. After fractionation of the products with Amberlite XAD-2 and Sephadex LH-20 three main feruloylated oligosaccharides (F3-F7) were isolated. They represented approximately 30% of the ferulic acid, and approximately 2% of the neutral sugars contained in the hydrolysis supernatant. The compositions of F7, F6, and F3 were Fe-Ara (1:1), Fe-Ara-Xyl (1:1:1), and Fe-Ara-Xyl-Gal (1:1:1:1), respectively. The structures of the three oligomers were determined using chemical methods (methylation, acetalation, reduction) and 13C NMR spectroscopy: F7 was 5-O-(trans-feruloyl)-L-Araf;F6 was O-beta-D-Xyl p-(1-->2)-[5 O-(trans-feruloyl)-L-Araf]; and F3 was O-L-Gal p-(1-->4)-O-D-Xyl p-(1-->2)-[5-O (trans-feruloyl)-L- Araf]. F7 has been previously isolated from other monocots especially from wheat bran and soluble arabinoxylans from wheat flour; this is the first report of feruloylated oligosaccharides F6 and F3. Our results suggest that these oligomers are side-chain constituents of heteroxylans in maize bran. Ferulic acid is probably partly responsible for the insolubility of heteroxylans by coupling polysaccharide chains through ferulic acid dimers. PMID- 7497482 TI - A perspective: sound--an Aladdin's cave? PMID- 7497485 TI - Coronary dissection resulting from angioplasty with slow oscillating vs. rapid inflation and slow vs. rapid deflation. AB - To determine if slow inflation or slow deflation (compared to standard rapid inflation/deflation) would minimize coronary arterial dissection, we randomized 162 lesions from 136 patients undergoing coronary angioplasty with polyolefin copolymer balloons to slow oscillating (1 atmosphere/20 sec) vs. rapid (over 30 sec) inflation and slow (over 15 sec) vs. rapid deflation. The incidence of any dissection was nearly identical in the four inflation/deflation groups. The incidence of severe dissection, however, was significantly higher for the slow inflation/slow deflation group compared to the other three groups (38% vs. 15%, P = .024). For angioplasty performed with polyolefin copolymer balloons, slow deflation combined with slow oscillating inflation is associated with more frequent severe dissections. PMID- 7497483 TI - Intracoronary thrombus: still a risk factor for PTCA failure? AB - Pre-existing intracoronary thrombus has been associated with an increased risk of percutaneous transluminal coronary angioplasty (PTCA) failure. Whether intracoronary thrombus is an independent risk factor for failure is uncertain, as conflicting data exist in the literature. Additionally, given advances in patient selection and angioplasty balloon design, it is uncertain whether the current risk posed by intracoronary thrombus is as substantial as that in the early angioplasty experience. The primary objective of this study was to first assess whether pre-existing coronary thrombus was an independent predictor of angioplasty failure and if so, whether the risk due to thrombus had changed from the early angioplasty experience to the present time. Our prospectively collected angioplasty data base was used to identify individuals undergoing single-vessel angioplasty of a thrombus-containing segment from January 1, 1984 through December 1, 1991. Univariate and multivariate stepwise logistic regression techniques were utilized to analyze clinical, angiographic, and procedural characteristics associated with angioplasty failure. The study period was divided into three separate time periods and these used as variables in our multivariate analysis. In the study population that consisted of 2,699 patients with single vessel angioplasty, univariate analysis demonstrated that among many factors, thrombus was importantly associated with angioplasty failure (P < 0.0001). A multivariate logistic model of angioplasty failure was developed and thrombus achieved independent predictive significance in this model.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7497486 TI - A single center randomized trial assessing use of a vascular hemostasis device vs. conventional manual compression following PTCA: what are the potential resource savings? AB - We undertook a randomized controlled trial comparing VasoSeal, a collagen vascular hemostasis device (VHD), with manual compression to assess its role and potential cost savings in the PTCA population. Of 460 patients, 359 were excluded due to clinical instability (30%), groin problems (18%), suboptimal PTCA result (15%), and other reasons (37%). The remaining 101 patients were randomized to either VHD (51) or manual compression (50). Hemostasis time, time to ambulation, duration of hospital stay, and nursing time and intensity were significantly reduced in the VHD group. There were no major groin complications in either treatment arm but there was a trend toward more minor groin complications in the VHD patients. The application of VasoSeal reduced resource use in this randomized study and may translate into significant cost reductions in the general coronary angioplasty population. PMID- 7497487 TI - Cardiac angiosarcoma arising in a coronary artery: angiographic and pathologic findings. AB - We discuss a unique case of an angiosarcoma which arose in an atherosclerotic coronary artery. It was widely metastatic. We describe the angiographic findings and their differential diagnosis, and briefly discuss this relatively common cardiac malignancy. PMID- 7497488 TI - Severe mitral incompetence following balloon mitral valvuloplasty: complete resolution during follow-up. AB - Mitral incompetence (MR) is a complication of balloon mitral valvuloplasty. There are few reports of long-term outcome. We believe this is the first report in the literature of complete resolution during follow-up of severe mitral regurgitation resulting from balloon valvuloplasty. PMID- 7497489 TI - The "dented bladder": diagnosis of a retroperitoneal hematoma. AB - Retroperitoneal hemorrhage and associated hematoma is a rare but potentially life threatening complication of cardiac catheterization and coronary artery interventions. This case presents the potential diagnostic utility of a supine film of the abdomen for early identification of a retroperitoneal hematoma in a patient following acute infarction PTCA. PMID- 7497484 TI - Clinical experience with intracoronary tissue plasminogen activator: results of a multicenter registry. Intracoronary t-PA Registry Investigators. AB - Clinical experience with the use of intracoronary tissue plasminogen activator (t PA) is limited. We therefore undertook this study to document current clinical usage of intracoronary t-PA during a 2-yr period in a multicenter registry. Intracoronary t-PA was utilized on 206 occasions in 198 patients (154 men and 44 women; mean age, 59 +/- 12 yr). The mean dose of intracoronary t-PA was 31 +/- 15 mg. Indications for use included acute myocardial infarction (MI) (n = 83), preexisting thrombus with (n = 49) or without (n = 41) percutaneous transluminal coronary angioplasty (PTCA), unstable angina (n = 14), abrupt vessel closure (n = 11), and post-PTCA "clean-up" (n = 8). The Thrombolysis in Myocardial Infarction (TIMI Phase I) criteria were used to assess perfusion and degree of thrombus formation. Overall, the mean TIMI flow grade increased from 1.2 +/- 1.1 before treatment to 2.3 +/- 1.0 after treatment (P < 0.0001); the mean TIMI thrombus grade decreased from 3.2 +/- 1.0 before treatment to 1.6 +/- 1.4 after treatment (P < 0.0001). Complications included bleeding (9.2%), MI (17.6%), need for coronary artery bypass grafting (CABG) (9.2%), need for repeat PTCA/atherectomy/stents (4.9%), and ventricular fibrillation (1.7%, all associated with opening totally occluded vessels). There were 14 subsequent in hospital deaths: 13 of the patients who died had originally presented with MI; the other had experienced abrupt vessel closure during a PTCA procedure. Intracoronary t-PA appears to be effective in improving distal flow and decreasing thrombus burden; however, intracoronary delivery of t-PA has associated risks.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7497490 TI - A dual blood supply protects the right gastroepiploic arterial graft: report of two cases. AB - We report two cases whose right gastroepiploic arterial bypass grafts were protected by a dual blood supply due to the anatomical anastomosis between the celiac and the superior mesenteric artery systems. This unique characteristic of the graft adds an advantage for its use as a graft conduit. PMID- 7497491 TI - Successful repair of combined septal and biventricular free wall rupture after acute myocardial infarction. AB - The following case is the first report of successful repair of combined left and right ventricular free wall and ventricular septal rupture. Reports of left ventricular free wall and septal rupture and reports of right ventricular free wall and septal rupture have been published both with and without successful surgical repair. This case demonstrates the presence of left and right ventricular free wall rupture along with septal rupture based upon cardiac catheterization and operative findings. In spite of her severe mechanical complications and prolonged hospital course, the patient is completely functional 1 year after surgical repair. PMID- 7497492 TI - Coronary artery fistula coexisting with a ventricular septal defect due to a penetrating gunshot wound. AB - We describe a patient who survived a penetrating gunshot wound to the chest. Cardiac evaluation including echocardiography and cardiac catheterization revealed a coronary artery fistula and a ventricular septal defect. This combination has been rarely reported in patients with penetrating cardiac injury. Follow-up echocardiography performed 15 mo later showed persistent left-to-right shunt. This is the first case in the literature demonstrating the natural history of combined coronary artery fistula and ventricular septal defect without surgical correction. PMID- 7497493 TI - Interventional physiology. Part XIII: Role of large pectoralis branch artery in flow through a patent left internal mammary artery conduit. PMID- 7497494 TI - Repeat embolization of the side branch of the internal mammary artery graft by gelatin sponge particles and micro coils. AB - The internal mammary artery (IMA) is increasingly used as a coronary bypass conduit because of better long-term patency and improved prognosis as compared with venous grafts. Previous investigators have suggested that the "steal" of blood flow of the IMA graft via the subclavian artery or a persistent large side branch of the graft may lead to its thinning. However, only a few reports have described the embolization of a large side branch using a transcatheter procedure. We present a case of repeated embolization of a large lateral costal side branch of the left internal mammary arterial graft applying gelatin sponge particles and micro coils, as well as angioplasty to the graft conduit, with resulting easing of chest pain. PMID- 7497495 TI - Intravascular ultrasound in the management of acute coronary artery dissection: a case report. AB - Crossing total occlusions is frequently difficult. The guidewire may enter a false lumen, thereby preventing successful balloon dilatations. We present a case of an acute arterial dissection following attempted angioplasty of a totally occluded right coronary artery. With an intravascular ultrasound probe in the false lumen, we were able to visualize a second guidewire and direct its passage into the true arterial lumen. This allowed for successful balloon dilatation and stent deployment restoring vessel patency. PMID- 7497496 TI - Dual guiding catheters for saphenous venous graft protection during native coronary angioplasty. AB - The simultaneous use of two guiding catheters during angioplasty has been previously described for native coronary artery bifurcation lesions. We describe a case in which a dual guiding catheter, dual guidewire technique is used for angioplasty of a native vessel lesion spanning the anastomotic site of a sequential saphenous vein graft. PMID- 7497498 TI - Localized drug delivery in atherosclerotic arteries via a new balloon angioplasty catheter with intramural channels for simultaneous local drug delivery. AB - A dual-purpose angioplasty catheter with intramural channels and exterior pores for local drug delivery ("channeled balloon") was studied in eight atherosclerotic human necropsy arteries and in 22 rabbits with atherosclerotic peripheral arteries, in which markers (0.005 microns to 15 microns) were infused locally at 2 atmospheres during simultaneous angioplasty at 6 atmospheres. Thirteen of the rabbits were sacrificed at 4 or 24 h after procedure to determine the intramural retention over time. Histology confirmed effective angioplasty and revealed presence of markers in the arterial wall in 29 of 43 treated arteries (67%), whereas all control segments without local delivery had no marker staining. majority of the ineffective local delivery (12/14) occurred when 15 micron particles were infused (12/13 arteries without intramural markers), especially when examined 4 or 24 h later. Thus, in atherosclerotic arteries, the channeled balloon enabled simultaneous local drug delivery at low pressure during effective angioplasty, although particle size may play a role in successful intramural impregnation and retention. PMID- 7497497 TI - Retrograde pulmonary vein wedge angiography: a new technique. AB - A new technique of pulmonary vein wedge angiography to delineate pulmonary artery anatomy is described. Conventional pulmonary vein wedge angiography requires an interatrial communication--natural or created by a transseptal puncture. In retrograde pulmonary vein wedge angiography, the left atrium and pulmonary veins are entered from arterial route using a specially designed catheter. This technique can be specially utilized for patients in whom there is no interatrial communication. PMID- 7497501 TI - In-hospital outcome of percutaneous transluminal coronary angioplasty for long lesions and diffuse coronary artery disease. AB - We performed coronary angioplasty on 151 long or diffuse lesions (120 long and 31 diffuse) in 141 patients (86% male, mean age 50 +/- 9 years). Long lesions were defined as lesions 11-20 mm in length and diffuse lesions as lesions longer than 20 mm, or three or more lesions in the same vessel. One or more adverse morphologic features were present in 131 (93%) lesions. Long balloons were used in 44%, significantly more often for diffuse disease (long lesions 39% and diffuse disease 64%; P = 0.004). Newer devices including the rotational atherectomy device (9 lesions), stents and perfusion balloons were employed in 18 (12%) lesions, more often for diffuse lesions (long lesions 8% vs. diffuse lesions 26%; P = 0.017). Lesion severity was comparable in the two groups (long lesions: 88 +/- 7%; diffuse lesions: 88 +/- 8%), but diffuse lesions were associated with significantly higher residual stenosis (long lesions: 6 +/- 8%; diffuse lesions: 12 +/- 13%, P = 0.01). Major complications occurred in five (3.5%) patients, including one death (0.7% mortality). The angiographic and clinical success rates for all patients were 99% and 96%, respectively, and were comparable for long and diffuse lesions. Judicious case selection and the use of long balloons and newer interventional devices permit coronary angioplasty for long lesions and diffuse disease with excellent success and a low risk of complications. Diffuse lesions are associated with more frequent use of long balloons and newer devices, especially rotational atherectomy and slightly higher residual stenosis as compared to long lesions. PMID- 7497500 TI - Clinical effects and utility of intracoronary diltiazem. AB - Coronary artery spasm is a known complication of coronary interventions, for which intracoronary nitroglycerin (ICN) is the treatment of choice. Some forms of intense spasm are resistant to ICN. Calcium channel antagonists are also known to be effective for coronary artery spasm, including nitroglycerin-resistant spasm. Here we describe a protocol for the clinical use of intracoronary diltiazem (ICD). By this protocol, ICD can be safely given without disturbing the clinical status of patients. ICD (2.5 mg) given slowly over 1 minute produced no vasodilitation of normal vessel segments but did produce significant dilatation of stenotic segments above and beyond the effects of nitrates. Mean minimum lumen diameter increased 18%, from 0.89 +/- 0.06 mm to 1.06 +/- 0.07 mm (mean +/- SEM, P < 0.001). ICD produced clinically insignificant changes in systolic blood pressure, diastolic blood pressure, heart rate, and PR, QRS, and QT intervals. This protocol has been employed to safely use ICD to relieve both nitroglycerin resistant epicardial artery spasm and nitroglycerin-resistant distal microvascular spasm (the no-reflow phenomenon). PMID- 7497499 TI - Kinetics of drug delivery to the arterial wall via polyurethane-coated removable nitinol stent: comparative study of two drugs. AB - Polymer-coated removable stents were used to deliver 14C-labeled etretinate and 3H-labeled forskolin to the vessel wall in 31 New Zealand White rabbits to study their kinetics. Stents loaded with etretinate (n = 8) and forskolin (n = 14) were implanted in the rabbit carotid arteries, and the animals were euthanized at different time intervals. Drug levels were measured in the media and adventitia of the stented segment, in distant tissues, and in blood. In four rabbits, forskolin-loaded stents were percutaneously retrieved 2 hr after implantation in the carotid artery, and the tissue and blood levels were determined 2 and 24 hr after retrieval. In seven rabbits etretinate-loaded stents were retrieved 72 hr after implantation in abdominal aorta, and drug levels were measured in the tissues and blood immediately after and at 1 and 4 days after retrieval. Levels of etretinate in the vessel wall peaked at 24 hr (250 ng/mg) and remained high up to 72 hr (185 ng/mg) after stent placement. Levels of forskolin peaked within 2 hr of stent placement (135 ng/mg) and rapidly declined to 4.9 ng/mg at 24 hr with the stent in situ. About 50% (1.4 mg) of the original etretinate remained in the stent at 72 hr compared to about 5% (0.08 mg) of forskolin at 24 hr. Ratio of peak drug levels in the vessel wall to those in the blood was 6,000 for etretinate and 780 for forskolin. (ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7497503 TI - Balloon impasse: a marker for severe mitral subvalvular disease and a predictor of mitral regurgitation in Inoue-balloon percutaneous transvenous mitral commissurotomy. AB - In certain instances of percutaneous transvenous mitral commissurotomy, the Inoue catheter balloon, although deflated and properly aligned, becomes held up or checked at the mitral valve. This "balloon impasse," observed in 13 of 760 patients undergoing the commissurotomy, reflects severe obstructive subvalvular disease even though echocardiographic evidence suggests otherwise. Our experience shows that the sign portends severe mitral regurgitation if the usual balloon sizing method is used. Such a situation occurred with four of the first six patients. In the next seven patients, the use of smaller balloon catheters (PTMC 18 or PTMC-20) for the initial set of stepwise dilatations averted creation of severe mitral regurgitation. When the "balloon impasse" sign is encountered during the commissurotomy procedure, the catheter selection and balloon sizing method should be judiciously altered. PMID- 7497502 TI - Cost comparison between two modes of Palmaz Schatz coronary stent implantation: transradial bare stent technique vs. transfemoral sheath-protected stent technique. AB - Coronary Palmaz Schatz stent implantation is usually performed by using the sheath protected stent delivery system (SDS) via the percutaneous transfemoral route. However, downsizing of PTCA equipment made transradial coronary stenting feasible. Bare stent implantation, 6F technique, increased patient mobility, reduced vascular complications and reduced hospital stay may increase cost effectiveness of this novel technique. Two well-documented patient groups selected for elective single vessel and single lesion Palmaz Schatz stent implantation were retrospectively compared. Group A (transradial stenting; n = 35) was compared to Group B (transfemoral stenting; n = 25) derived from the Benestent population, included in our hospital. A comparison was made for three areas of interest: (1) procedural consumption of material (the number of guiding catheters, guidewires, balloon catheters and stents), (2) postprocedural need for diagnostic and therapeutic procedures for stent-related complications, and (3) duration of hospital stay. Differences between these subjects in Group A and B were translated to hospital costs. Although more guiding catheters were used in group A (1.69 +/- 0.87 vs. 1.08 +/- 0.28; P = 0.001), the use of the SDS contributed importantly to higher material costs in group B (cost reduction in group A; 13%). Less patients in group A required diagnostic (2 vs. 7; P = 0.027) and therapeutic (0 vs. 5; P = 0.01) procedures for bleeding complications (cost reduction; 93%). Hospitalization in Group A was shorter (6.4 +/- 4.7 vs. 11.6 +/- 9.9 days; P = 0.005), caused by early and safe mobilization, less vascular complications, and preprocedural adjustment on coumadin (cost reduction; 45%).(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7497504 TI - Congenital clavicular pseudoarthrosis associated with vascular thoracic outlet syndrome: case presentation and review of the literature. AB - Thoracic outlet syndrome (TOS) associated with congenital clavicular pseudoarthrosis is rare in adults and often misdiagnosed. In this case report, we describe an adult female who was found to have thrombosis of the subclavian and axillary arteries with embolization documented by invasive angiography. This unusual vascular manifestation of TOS should remind physicians that anatomic derangements may predispose to upper extremity ischemia. PMID- 7497505 TI - Combined dissection of the right coronary artery and right coronary cusp during coronary angiography. AB - Dissection of coronary arteries during diagnostic coronary angiography is infrequent; dissection of the coronary cusp is extremely rare. The combination of coronary artery and coronary cusp dissection has not been reported previously. A case of right coronary artery dissection and right coronary cusp dissection during diagnostic coronary angiography is described. PMID- 7497507 TI - Assessment of transplant arteriopathy by intracoronary two-dimensional ultrasound imaging and coronary flow velocity. AB - Transplant coronary arteriopathy is the major impediment to the long-term survival of cardiac allografts. This report highlights two-dimensional imaging and Doppler flow to assess transplant coronary arteriopathy. PMID- 7497508 TI - Coronary angioplasty of bifurcational lesions with the protecting branch technique using 6 French guiding catheter. AB - The use of 6 French guiding catheters is still usually limited to elective percutaneous transluminal coronary angioplasty. We describe our preliminary experience with this guiding size to perform a double guide wire procedure for angioplasty or coronary bifurcation lesions, to protect (n = 5) and/or to sequentially dilate (n = 9) a major side branch. Technical success was 12/14 (86%), angiographic success 13/14 (93%), and in-hospital outcome was event free in all patients. Thus, 6 French guiding size is both safe and effective for the majority of coronary bifurcation angioplasty. PMID- 7497509 TI - A novel approach to the placement of Palmaz-Schatz biliary stents in saphenous vein grafts. AB - The placement of Palmaz-Schatz biliary stents has become a successful method to treat stenoses in large saphenous vein grafts. Unlike coronary stents, the biliary stents are not routinely delivered with a selective delivery sheath and may be more difficult to deliver and prone to detachment from the balloon and even embolization during delivery. In order to enhance the ability to deliver these stents, a guide-within-a-guide system was developed. A 7F guiding catheter was used as a selective delivery sheath within a standard guiding catheter. Nineteen biliary stents were successfully placed in vein grafts in 15 patients using this system. One procedure was complicated by an embolic event documented angiographically following intragraft delivery of urokinase but prior to stent implantation. PMID- 7497506 TI - Left ventriculography complicated by cerebral air embolism. AB - Cerebral air embolism is a severe complication of various invasive medical procedures. Hyperbaric oxygen is the sole definitive therapy. We describe a 68 year-old patient who presented with upper left limb motor and sensory deficits following the injection of a contrast medium for left ventriculography. Numerous air bubbles were seen on cineangiography concomitantly with injection of the contrast medium. Immediate compression with hyperbaric oxygen resulted in complete resolution of all neurological symptoms. Iatrogenic cerebral air embolism is an underdiagnosed condition that may result from cardiac catheterization. Physician awareness will improve prevention, and prompt diagnosis and the use of hyperbaric oxygen will result in an optimal outcome if it does occur. PMID- 7497510 TI - Balloon expandable stent implantation after iatrogenic arterial dissection of the left subclavian artery. AB - A patient undergoing attempted angioplasty of a left internal mammary artery graft to left anterior descending coronary artery developed an iatrogenic dissection of the left subclavian artery, with symptoms of left arm claudication. A balloon expandable stent was inserted with an excellent angiographic result. The patient has remained free of symptoms or signs of decreased arm flow, with a patent subclavian artery demonstrated on angiography 8 months later. The excellent early-to-moderate term clinical and angiographic results support the efficacy of this technical approach. PMID- 7497511 TI - Left heart catheterization by direct puncture with two-dimensional echocardiographic guidance: a case report. AB - In patients with mechanical valves in both the mitral and aortic positions, direct ventricular puncture is the recommended approach to catheterization. We report a case in which two-dimensional (2D) echocardiography was used to guide left heart catheterization by direct ventricular puncture and propose that the use of 2D echocardiography might reduce the risks historically associated the procedure. PMID- 7497512 TI - Assessment of collateral flow during balloon coronary occlusion by intracoronary blood flow velocity analysis. AB - Coronary blood flow velocity analysis of the contralateral and of the ipsilateral artery can be used to assess collateral flow during controlled balloon coronary occlusion. Coronary blood flow velocity assessment in combination with measurement of aortic pressure and coronary wedge pressure permits expression of the development of the collateral circulation in terms of flow and resistance. The application of both techniques is illustrated in two patients. PMID- 7497513 TI - Transluminal extraction catheter atherectomy in long saphenous vein grafts. AB - Lesions in distal coronary vasculature present unique problems for catheter-based interventions. We report two cases of attempted transluminal extraction catheter (TEC) atherectomy for very distal lesions in long circular saphenous vein grafts. One case succeeded with a simple modification of the TEC device; another failed because of excessive tortuosity in the proximal graft. This experience helps define the potential and limitations of TEC. PMID- 7497514 TI - Anterograde perfusion in acute limb ischemia secondary to vascular occlusive cardiopulmonary support. AB - Cardiopulmonary support (CPS) can resuscitate a patient with circulatory collapse during high-risk interventional procedures, although vascular complications may accompany its use. We report a patient with cardiogenic shock secondary to myocardial infarction who required extended CPS support associated with acute infarct-related coronary artery angioplasty and stent placement. Leg ischemia due to an occlusive cannula was resolved using a percutaneous anterograde perfusion device. In general, such devices may have application in patients dependent on mechanical support associated with limb ischemia. PMID- 7497515 TI - Angioplasty may be contraindicated in occlusions which are composed mainly of thrombus. PMID- 7497516 TI - "Dancing needle sign" for obtaining access to the femoral artery. PMID- 7497518 TI - Preferential expression of V beta gene families in CD8 memory cells of apparently healthy donors. AB - Antigen stimulation may lead to expansion or deletion of T-cells expressing T cell receptors that belong to specific V beta gene families. Since such stimulation at the same time will lead to conversion from naive to memory T cells, we have asked whether a bias in V beta families can be observed when comparing these two populations. We have studied the expression of V beta 3, 8, 13.3, 19, and 22 in peripheral blood T-cells for 12 apparently healthy male donors. For flow cytometry 100,000 CD4+ or CD8+ cells each were analysed in three colour immunofluorescence for percentage of V beta families among CD45R0- naive and CD45R0+ memory cell. Greater than twofold excess was found in the CD8+CD45R0+ cells in four cases (1 x V beta 13.3, 2 x V beta 19, 1 x V beta 22) and a greater than twofold decrease in CD8+CD45R0+ cells in two cases (1 x V beta 8, 1 x V beta 22). In contrast, among CD4+ cells no such bias was detected. The excess in CD8+CD45R0+ memory cells showed no substantial fluctuation over time in that it was found to be stable for 19 to 70 days. Finally, in vitro conversion of purified CD8+CD45R0- cells to CD45R0+ cells by polyclonal stimulation with PHA did not result in the excess of V beta usage observed in vivo. These data suggest that specific antigen stimulation during past infection or allergy may be responsible for the excess of certain V beta gene families. Clinical studies looking for disease associations will have to test CD4 and CD8 naive and memory subsets in order to precisely identify a bias in V beta usage and these studies will have to consider the pronounced changes observed in healthy controls. PMID- 7497517 TI - New modular system for the determination of coronary blood flow velocity. PMID- 7497519 TI - Intermediate TCR cells with self-reactive clones are effector cells which induce syngeneic graft-versus-host disease in mice. AB - It has been established that, even after syngeneic bone marrow transplantation, animals treated with immunosuppressive drugs may suffer from graft-versus-host disease, showing autoimmune-like symptoms. Although the major effector cells are known to be T-cell subsets, detailed characterization of such T cells remains to be investigated. In the present study, we characterized them, especially as to whether they are thymus-derived T cells or extrathymic T cells, and how self reactive clones were distributed among the above T-cell subsets. BALB/c mice (Mls 1b2a) were irradiated (9 Gy), subjected to bone marrow transplantation, and then treated with cyclosporin A (CsA) for 6 weeks. From 2 weeks after the cessation of CsA, these mice displayed signs of GVH disease. The major target organs included the liver and colon. Two-color staining for CD3 and IL-2R beta was applied to identify CD3-IL-2R beta+ NK cells, CD3-intermediate +IL-2R beta+ cells (i.e., intermediate CD3 or TCR cells of extrathymic origin) and CD3-high+IL-2R beta- cells (i.e., high CD3 cells of thymic origin). It was demonstrated that the major expanding lymphocytes were intermediate TCR cells and that self-reactive clones (V beta 3+ and V beta 11+ cells in this strain of mice) were confined to this population. Interestingly, these self-reactive clones had ability to respond to immobilized anti-V beta 3 and anti-V beta 11 mAbs. Liver MNC in mice with GVH disease which contained the highest proportion of intermediate TCR cells were able to mediate the adoptive transfer of GVH disease to other irradiated (6.5 Gy) mice. Intermediate TCR cells also showed potent cytotoxic activity against syngeneic leukemia cells. These results suggest that intermediate TCR cells are the major effector cells for the induction of syngeneic GVH disease. PMID- 7497520 TI - Antigen-induced death of alloreactive human T-lymphocytes occurs in the absence of low molecular weight DNA fragmentation. AB - Stimulation via the CD3/TCR molecular complex induces proliferation of resting T cells, but triggers programmed cell death (apoptosis) in immature thymocytes and preactivated mature T cells. Activation-induced cell death (AICD) triggered by anti-CD3/TCR mAb or by staphylococcus enterotoxin superantigen is associated with fragmentation of genomic DNA into oligonucleosomal fragments of 200 bp length, thus displaying the characteristic features of apoptosis. Here, we show that a fraction (20-50%) of cells in alloreactive CD8 human short-term T cell lines, generated by repeated restimulation with EBV-transformed B cell lines, undergo AICD when restimulated with the appropriate (but not with third party) stimulator cells. AICD of responder T cells is inhibited when stimulator cells are preincubated with anti-HLA class I mAb but not with anti-HLA class II mAb, indicating that T cell death is dependent on alloantigen (HLA class I) recognition by responding CD8 T cells. Importantly, alloantigen-induced T cell death occurs in the absence of detectable DNA fragmentation. Thus, several independent assay systems all failed to reveal low molecular weight DNA fragmentation, even though DNA fragmentation was readily detected in T cell lines exposed to PHA or gamma-irradiation. Alloantigen-induced T cell death was prevented by aurintricarboxylic acid, which has previously been shown to inhibit apoptosis in experimental systems where no DNA fragmentation occurs. Taken together, these results demonstrate that alloantigen can trigger AICD in mature responding T cells in the absence of low molecular weight DNA fragmentation. PMID- 7497521 TI - Suppression of collagen-induced arthritis by an angiogenesis inhibitor, AGM-1470, in combination with cyclosporin: reduction of vascular endothelial growth factor (VEGF). AB - Pannus formation characterized by neovascularization is a prominent pathologic finding in both rheumatoid arthritis (RA) and rat collagen-induced arthritis (CIA). CIA is a T-cell-dependent process induced by immunization of inbred LOU rats with native type II collagen in incomplete Freund's adjuvant. AGM-1470 is a highly specific inhibitor of new blood vessel formation by its effects on endothelial cell migration, endothelial cell proliferation, and capillary tube formation. Cyclosporin A (CSA) is an immunomodulating agent that inhibits IL-2 and other cytokine production involved in early antigen activation of T-cells. In this study the effects of single and combination therapy with AGM-1470 (27 mg/kg alternate days) and low-dose CSA (4 mg/kg/day continuous infusion via osmotic pump) on established CIA (total n = 62) were examined. At Day 18 post arthritis onset, clinical arthritis was significantly reduced in rats treated with single agent AGM-1470 (1.88 +/- 0.33) or combination therapy (1.13 +/- 0.32) (P < 0.00001 and 0.000001, respectively) versus control. Single-agent CSA-treated rats, even if given CSA beginning on the day of immunization, did not attenuate arthritis severity. THe longitudinal mean arthritis score of combination-treated rats was significantly lower than that of rats receiving AGM-1470 (P < 0.0001), reflecting a more moderate early disease course in combination-treated rats. Disease severity in rats treated with single-agent CSA was not significantly different from control rats. Mean WBC counts, differentials, and delayed-type hypersensitivity responses were similar in all groups. CII antibody levels were lower in AGM-1470 protocols compared to CSA or controls. Flow cytometry of peripheral blood, spleen, and lymph nodes demonstrated decreased levels of CD4+ cells in rats given CSA. TNF-alpha levels remained elevated, even in treated rats, while vascular endothelial growth factor levels were reduced in rats receiving AGM-1470 compared to both arthritic controls and naive rats. Both single-agent and combination therapies were well tolerated. This is the first study to examine the effects of AGM-1470 together with CSA. Combination therapy was more effective than single-agent therapy. The results suggest that the use of interventions with distinct mechanisms of action may be efficacious in the treatment of RA. PMID- 7497524 TI - Superantigen reactivity of gamma delta T cell clones isolated from patients with multiple sclerosis and controls. AB - gamma delta T cells have been implicated as playing a role in the demyelinating processes of multiple sclerosis (MS). However, the nature of the ligands which lead to activation and accumulation of gamma delta T cells in the brain lesions remains unknown. This study was undertaken to examine whether gamma delta T cells derived from cerebrospinal fluid and blood of MS patients could be stimulated by bacterial superantigens: staphylococcal enterotoxins A and B and toxic shock syndrome toxin-1. A panel of 16 gamma delta T cell clones isolated from MS patients and controls was found to react with the superantigens used at a nanogram range and displayed specific cytotoxic activity toward target cells pulsed with the corresponding superantigens. The responses of the gamma delta T cell clones did not require MHC-matched accessory cells and were not blocked by antibodies to the MHC molecules, suggesting a non-MHC restricted interaction. The superantigen reactivity was associated with both V delta 2+/V gamma 2+ and V delta 1+/V gamma 1+ subsets, reportedly found in the MS lesions. Our data suggest an alternative pathway which may account for gamma delta T cell activation in MS. PMID- 7497522 TI - Sequence elements required for function of the MHC class II A beta gene promoter in murine macrophages. AB - We have studied the DNA sequence elements of the murine MHC class II A beta gene involved in transcriptional regulation in macrophages. For this study, the A beta promoter was fused to the human growth hormone gene and transfected into bone marrow-derived macrophages. These macrophages were stimulated by interferon-gamma (IFN-gamma), after which cellular RNA was assayed for the amount of human growth hormone transcripts. A -146 to +14 fragment of the A beta promoter was found to be sufficient to confer positive regulation by IFN-gamma. Induction was suppressed by bacterial lipopolysaccharide, tumor necrosis factor (TNF-alpha), or maley-lated bovine serum albumin. Substitution mutations were made within each of the conserved sequence elements of the promoter as well as within the spacer regions between these elements. All four conserved sequences found in class II promoters, the H, X1, X2, and Y elements, were found to be essential for promoter activation in macrophages. PMID- 7497523 TI - Suppression of immunopathology in schistosomiasis by interleukin-2-targeted fusion toxin, DAB389IL-2. I. Studies of in vitro and in vivo efficacy. AB - Schistosomiasis causes pathology in an estimated 200 million individuals. Clinical disease is caused by a complex immunopathologic response to the parasite ova, which are deposited in the host tissues. This immunopathologic response is caused by T lymphocytes which express the high-affinity IL-2 receptor (IL-2R). DAB389IL-2 is a diphtheria toxin-IL-2 fusion toxin protein which functionally inactivates or kills cells which bear the high-affinity IL-2R. DAB389IL-2 has been used in man to suppress IL-2R-dependent immune reactivity. Therefore, we reasoned that DAB389IL-2 might suppress immunopathology in schistosomiasis. In these studies we assessed the in vitro and in vivo effects of DAB389IL-2 on the development of immunopathology in murine schistosomiasis. DAB389IL-2 suppressed IL-2, lectin mitogen (Con A), and soluble Schistosoma mansoni egg antigen-induced lymphocyte proliferation and in vitro granuloma formation. In addition, DA-B389IL 2 suppressed in vitro IL-2R expression. DA-B389IL-2 also suppressed the development of granulomas and collagen deposition in vivo in the livers of infected animals. Therefore, DAB389IL-2 may have potential for the targeted reduction of immunopathology due to schistosomiasis in man. PMID- 7497526 TI - Endogenous cytokine expression profiles in retinoic acid-induced IgA production by LPS-stimulated murine splenocytes. AB - All-trans-retinoic acid (RA) enhances IgA production by LPS-stimulated murine splenocytes. After stimulation by RA and LPS, or by LPS alone, total RNA was extracted from cultured cells on Days 1 to 4, and the kinetics of expression of various cytokine mRNAs were analyzed by the RT-PCR method. RA induced the expression of IL-5 and TGF-beta 2 mRNAs in the LPS-stimulated cells. In addition, the expression of IL-6 and IL-2 mRNAs was more intensive in RA-stimulated cells than in unstimulated cells. TGF-beta 1 and TGF-beta 3 mRNAs were constitutively expressed in both culture groups. RA enhanced IgA production by LPS-stimulated spleen cells but not that by LPS-stimulated mu(+) naive splenic B-cells. For RA induced IgA production, the B-cells required T-cells or the culture supernatant from RA-stimulated T-cells. Furthermore, exogenous IL-5 replaced the T-cell requirement, at least in part, in RA-induced IgA production by LPS-stimulated B cells. This reaction was partially inhibited by anti-TGF-beta-neutralizing antibodies. These findings suggest that RA induces IgA production by (IL-5 + LPS) stimulated B-cells in TGF-beta-independent and TGF-beta-dependent manners. PMID- 7497528 TI - HIV-induced syncytia in peripheral blood cell cultures crawl by extending giant pseudopods. AB - It was previously demonstrated that HIV-induced syncytia of the immortalized T cell line SupT1 reorganize their cytoskeleton and form a spherical supernuclear complex, thus mimicking the organization, polarity, and morphology of a single SupT1 cell. Then, through extension of a single, giant pseudopod, these syncytia, which grow to more than 100 times the volume of a single SupT1 cell, translocate along a substratum. To verify that syncytium motility is not peculiar to the SupT1 cell line, we have analyzed the cytoskeletal organization and motile capabilities of HIV-induced syncytia formed in peripheral blood cell cultures containing more than 90% CD4-positive cells. The results demonstrate that although peripheral blood T cells differ from SupT1 cells in size and morphology, they are continuously motile and translocate along a substratum in a manner quite similar to that of SupT1 cells, and peripheral blood T cell syncytia induced by HIV-1LAI as well as two additional clinical isolates translocate by the extension of a giant anterior pseudopod in a fashion indistinguishable from that of HIV induced SupT1 syncytia. Together, these results support the generalization that HIV-induced T cell syncytia are motile. PMID- 7497527 TI - Immunohistochemically defined lymphocyte function-associated antigen 1 (LFA-1) on an adenocarcinoma cell line is a distinct molecule from LFA-1 on leukocytes. AB - We previously reported that the immunohistochemically defined LFA-1 antigen (LFA 1-like antigen) was expressed on various exocrine tissues uninvolved with tumors in patients with malignant diseases using LFA-1 alpha-specific monoclonal antibodies (mAb) 2F12 and HVS6B6. In this study we investigated differences at the molecular level between LFA-1 on leukocytes and LFA-1-like antigen on MKN45.16, a subline derived from an adenocarcinoma line MKN45 that expresses a high amount of LFA-1-like antigen. LFA-1-like antigen was reactive to mAb 2F12 or HVS6B6, but not to any of the other five different LFA-1 alpha (CD11a)-specific or four LFA-1 beta (CD18)-specific mAb. mAb 2F12 immunoprecipitated a 200-kDa membrane protein (LFA-1-like antigen) from MKN45.16 cells, whereas it precipitated 180-kDa (LFA-1 alpha) and 95-kDa (LFA-1 beta) proteins from a monocytic cell line (THP-1) under both nonreducing and reducing conditions. The molecular difference was confirmed further by N-glycanase treatment of the immunoprecipitates. The isoelectric point of LFA-1-like antigen was 6.0, whereas those of LFA-1 alpha and LFA-1 beta were 6.0 and 4.7, respectively, by two dimensional gel electrophoresis. Expression of LFA-1 alpha gene on MKN45.16 cells was not detected at the mRNA level by six different sets of LFA-1 alpha-specific oligonucleotides and reverse transcription-polymerase chain reaction. These results indicated that LFA-1-like antigen on an adenocarcinoma cell line was a distinct molecule from LFA-1 on leukocytes. PMID- 7497525 TI - Fas involvement in cytotoxicity mediated by human NK cells. AB - Lysis of target cells (TC) by cytolytic lymphocytes involves the secretion of cytoplasmic granules containing perforin and serine esterases by the effector cell (EC). Recently, a granule-independent cytolytic mechanism involving the interaction of the apoptosis-triggering Fas antigen (CD95) with Fas ligand (FasL) has been revealed in T cells. However, whether the Fas lytic pathway also functions in NK cells has not been established. We purified human peripheral NK cells (> 98% CD56+) and found that PMA and ionomycin treatment upregulated FasL message and stimulated the NK cells to lyse a Fas+ TC. This lysis was partially inhibited by the anti-Fas-blocking antibody M3 or by Fas.Fc fusion protein. We also found that FasL is constitutively expressed on the human NK-like leukemia cell line YT-INDY and that YT-INDY utilizes a Ca(2+)-independent Fas lytic pathway, as well as the granule pathway. We have previously shown that CD28/B7 interactions are involved in TC recognition by YT-INDY. K562 cotransfected with Fas and B7-1 (K562/Fas/B7) was lysed by YT-INDY at a higher level than a vector transfected K562 line, whereas K562 transfected with Fas alone was not. Lysis of K562/Fas/B7 cotransfectants was partially Fas-mediated, as indicated by the presence of Ca(2+)-independent, M3-inhibitable lysis. Ca(2+)-independent, Fas mediated lysis of several TC by YT-INDY was inhibited by anti-CD28 antibody. Anti LFA-1 also inhibited Fas-mediated cytotoxicity in YT-INDY. Thus, fresh human NK cells and the human NK-like cell line YT-INDY are capable of using the Fas lytic pathway. In YT-INDY, CD28/B7 and LFA-1/ICAM interactions appear to influence the Fas lytic pathway. PMID- 7497529 TI - Adhesion molecules on intermediate TCR cells. II. Hepatoprotective effects of hyaluronic acid on acute liver injury. AB - The liver is a major organ wherein extrathymic T cells and NK cells exist in mice. Due to their unique properties, i.e., extrathymic T cells are TCR (or CD3) intermediate+ IL-2R beta+ (herein termed intermediate TCR cells) and NK cells are TCR(-)IL-2R beta+, they are easily distinguished from the other lymphocyte subsets by using mAbs in conjunction with immunofluorescence tests. They were recently found to express a higher level of CD44 antigen, which is a ligand for hyaluronic acid, than that of another T cell subset (i.e., thymus-derived T cells or bright TCR cells). Since an intravenous administration of hyaluronic acid was also found to reduce the number of intermediate TCR cells and NK cells in the liver, we examined whether hyaluronic acid had a hepatoprotective effect on acute liver injury. Such injury was induced by LPS injection in mice pretreated with Propionibacterium acnes 1 week earlier. When a single dose of hyaluronic acid was given to these mice 12 hr before LPS injection, a prominent hepatoprotective effect was observed in terms of decreases of mortality (up to 50%), lymphocyte infiltration of the liver, serum transaminase levels, and tissue damage. At this time, liver mononuclear cells isolated from the treated mice showed decreased levels of cytokine production such as TNF and IL-1. These results reveal that intermediate TCR cells and NK cells in the liver actually adhere the sinusoid walls by means of an interaction of CD44 molecules and hyaluronic acid even in the case of acute liver injury. It suggests a possible therapeutic effect of the administration of hyaluronic acid in acute liver injury by eliminating the effector cells and cytokine-producing cells from the liver. PMID- 7497531 TI - Myelin proteolipid protein-induced Th1 and Th2 clones express TCR with similar fine specificity for peptide and CDR3 homology despite diverse V beta usage. AB - Myelin-specific T-helper (Th) cells which induce encephalomyelitis belong to the inflammatory Th1 subset. Th2 cells recognizing similar epitopes potentially represent specific inhibitors of encephalitogenic Th1 cells. Since the differential stimulation of antigen-specific Th2 cells may be important in the regulation of autoimmune inflammatory disorders, we have examined the fine specificity of a Th1 and a Th2 clone, induced by immunization of SJL mice with native proteolipid protein (PLP) and specific for the PLP 139-151 sequence. Stimulation of the clones by synthetic peptides containing single alanine substitutions demonstrated that L141, W144, H147, and P148 represent critical residues. Surprisingly, this pattern was identical for both subsets. Competition studies indicated indirectly that L141 and P148 may be MHC-binding residues, whereas W144 and H147 contact the TCR. Sequencing of the TCR expressed by both Th subset clones demonstrated different V beta usage as well as variation in the D region sequence and length. Interestingly, realignment of the sequence of the CDR3 regions showed striking homology. This study demonstrates that Th1 and Th2 subsets can express very similar peptide specificities, while utilizing very different TCR V beta chains. These results suggest that the therapeutic modalities based on either peptide antagonists or antibodies specific for CDR3 may have limited effectiveness in treating autoimmune disorders, since they may also target the beneficial arm of the immune response. PMID- 7497530 TI - Substance P induces tumor necrosis factor in an ex vivo model system. AB - Many studies have shown that tumor necrosis factor (TNF) is a potent soluble mediator of immunoregulation and inflammation. Neuropeptide substance P (SP) has been known to exert significant influence on production of certain inflammatory cytokine by immune cells. Immunopathogenic mechanism underlying the effect of neuropeptide substance P (SP) and the specific amino acid sequence of SP that induces TNF has not been clearly studied. Employing ex vivo and in vitro model systems, we investigated the direct effect of different sequences of SP on TNF secretion by whole blood and separated total mononuclear cells. Aliquots of blood samples (1 ml) or Ficoll-Hypaque-separated total mononuclear cells (1 x 10(6)/ml) were cultured with different concentrations of SP and its sequences (SP 1-4, SP 4 11) or vasoactive intestinal peptide (VIP) for 24 hr at 37 degrees C. Plasma samples and culture supernatants were assayed for TNF levels in a bioassay using a TNF-sensitive WEHI 164 subclone 13 cell line. Plasma from blood samples or lymphocytes treated with whole SP and SP 4-11 at 10(-7), 10(-8), and 10(-9) M concentrations induced significant production of TNF compared to negligible levels of TNF produced by SP 1-4-treated and untreated cultures. VIP at all concentrations tested did not induce TNF production and was similar to untreated control cultures. Separated mononuclear cells also produced significant levels of TNF in response to SP and SP 4-11. Anti-TNF-alpha antibodies neutralized the TNF induced by SP 4-11 in plasma. These studies suggest that an ex vivo system using whole blood may be an ideal model to study the effects of SP on TNF production. These studies also demonstrated that the TNF inducing activity of SP residues in the region containing amino acids 4 to 11. PMID- 7497532 TI - Systemic lupus erythematosus. AB - Assessing patients with SLE is difficult because of the heterogeneity of the disease. The Revised 1982 ARA Classification Criteria set has been widely accepted for classifying SLE patients for inclusion in clinical studies, but it is not appropriate for making the diagnosis of SLE in an individual and is not helpful for classifying patients with early or mild disease in population based epidemiological studies. Further refinement of this criteria set to meet these objectives and to facilitate subdivision of patients with SLE into those with similar clinical, serological or genetic features poses a future challenge for the clinical epidemiologist. A number of valid and reliable indices are available for measuring clinical disease activity in SLE (Table 3). Despite their different approaches they have been shown to correlate well with each other and hence would appear to be measuring the same thing. The exact choice of instrument will be dictated by the purpose for which it is required. Although none is perfect they are useful for monitoring groups of patients in outcome studies in a research setting. Practically, disease activity indices are unlikely to be appropriate for dictating treatment decisions in individual cases: an instrument comprehensive and flexible enough for this purpose would undoubtedly be far too cumbersome for widespread use. Ultimately such fine tuning will continue to rely upon the skill and intuition of experienced physicians. PMID- 7497533 TI - Scleroderma. AB - In practice, the classification of scleroderma is less problematic than that of most other connective tissue diseases given the distinctive pattern of skin involvement. Classification of early and limited disease is more problematic and is difficult to separate from severe forms of Raynaud's phenomenon. Division of scleroderma into two groups depending on the presence or absence, early in the disease, of truncal skin involvement makes biological and clinical sense. Measurement using a clinical skin score remains the best approach to monitoring disease progress. PMID- 7497534 TI - Sjogren's syndrome. AB - The review describes the historical development and nomenclature of Sjogren's syndrome--primary and secondary. The strengths and weaknesses as well as similarities and dissimilarities for establishing the diagnosis of keratoconjunctivitis sicca and xerostomia used among the five most commonly classification criteria sets are focused upon in text and table. Validation of the different sets of criteria are performed. Primary and secondary disease activity markers and outcome measures as well as future research plans for improving diagnosis are dealt with. PMID- 7497535 TI - Inflammatory myopathies. AB - New information regarding myositis specific autoantibodies, histopathologic analysis of muscle biopsy specimens, and immunogenetic features of the different serologic subsets of disease has greatly increased our understanding of the pathogenesis of the inflammatory myopathies. The clinical descriptions of inclusion body myositis and 'amyopathic dermatomyositis' (Euwer and Sontheimer, 1993) are examples of our expanded descriptive capabilities in the evaluation of patients with myopathy. Finally, newer techniques such as cytokine analysis and magnetic resonance imaging may help in the ongoing assessment of disease activity in patients with myositis. The combination of these recently described clinical and laboratory parameters are enough to force a reconsideration of the previously described classification and diagnostic criteria in the inflammatory myopathies. PMID- 7497538 TI - Osteoporosis. AB - Two complementary notions need to be considered in the definition of osteoporosis: a reduction in bone mineral density and the occurrence of fracture. Bone mineral density is an established determinant of the risk of future fracture, but current interventions are most usefully targeted before bone density has fallen to the levels at which fractures usually occur. Bone density may now be measured accurately and precisely, and the sensitivity and specificity of this technology for fracture risk prediction are high. It therefore seems reasonable to classify osteoporosis in terms of both a reduction in bone density and the occurrence of fracture. On this basis, a four point scale has recently been proposed by the World Health Organization: 1) normal, 2) low bone mass, 3) osteoporosis and 4) established osteoporosis. While it is likely that this diagnostic classification will change as experience of osteoporosis increases, it is important to have widely adopted guidelines of this type which act as a framework for further research. PMID- 7497537 TI - Vasculitis. AB - The systemic vasculitides are a group of rare inflammatory conditions resulting in inflammation and necrosis of blood vessel walls. They are somewhat commoner than previously believed with an annual incidence approaching 40 per million. Furthermore the annual incidence of rheumatoid vasculitis is 12.5 per million and Wegener's granulomatosis is 8.5 per million. The first useful classification system for systemic vasculitis was published in 1952, since then a number of different schemes have been published. The major changes have been the recognition of the importance of dominant blood vessel size, the distinction between primary and secondary vasculitis and the incorporation of pathogenetic markers such as ANCA (see Table 6). Until relatively recently there were no widely agreed diagnostic or classification criteria. In 1990 the ACR published criteria for the diagnosis of polyarteritis nodosa, Churg-Strauss syndrome, Wegener's granulomatosis, hypersensitivity vasculitis, Henoch-Schonlein purpura, giant cell arteritis and Takayasu's arteritis. The criteria were provided in both traditional and tree format. Sensitivity and specificity rates varied considerably: 71.0-95.3% for sensitivity and 78.7-99.7% for specificity. The criteria were not tested against the general population or against patients with other connective tissue diseases or rheumatic conditions. In 1993/94 the Chapel Hill Consensus Conference developed and published definitions for the nomenclature of systemic vasculitis based on clinical features. These have not met with universal acceptance. However, they are a useful addition, since their use should result in different centres studying more homogeneous populations of patients and facilitate comparison of data between different centres. Assessment of vasculitis comprises an activity score (BVAS), damage index and quality of life/health status (SF-36). These are recent developments which are still undergoing validation. PMID- 7497536 TI - Polymyalgia rheumatica and giant cell arteritis. AB - The original descriptions of polymyalgia rheumatica (PMR) and giant cell arteritis (GCA) in the medical literature date back to 1888 and 1890, respectively. Classification criteria for PMR and GCA are not standardized since most authors used subjective criteria based on their personal experience. Only one study has evaluated criteria for PMR and has found seven variables with high discriminant value. Criteria for GCA are less varied because a positive biopsy of the temporal artery is diagnostic. However, combinations of different clinical and laboratory features have been used for diagnosis when biopsy is negative or missing. Assessment of PMR/GCA is based on the serial determination of markers of acute phase such as ESR, CRP, or plasma viscosity. However, their value in predicting recurrence of the diseases is poor. New immunological factors including soluble interleukin-2 receptors, interleukin-6, serum soluble CD8, and serum soluble intercellular adhesion molecule-1 are presently under investigation. PMID- 7497539 TI - Back pain. AB - Classification of back pain is a difficult task. Traditional schemes have focused on the small percentage of cases which have specific causes. Structural anomalies observed on X-ray examination explain only a small proportion of back pain cases, and the emphasis placed on these in the traditional schemes is, as Anderson put it, the tail wagging the dog (Anderson, 1977). Many syndrome classifications are based on arbitrary notions of cause, with little empirical justification and no evidence that they can reliably and usefully be applied in practice. More pragmatic approaches start with the separation of the serious from the less serious, and the distinction between spinal pain and pain arising from outside the spine. The classification of the large majority of back pain cases which are 'non-specific' is best approached by grading the severity of the clinical and psychological features of back pain and their disabling consequences. Such grading schemes also provide the most appropriate outcome measures for clinical and epidemiological back pain research. PMID- 7497541 TI - Fibromyalgia. AB - The article details the history, concept, definition and assessment of the still enigmatic condition of 'fibrositis' or, as it has more recently been called, 'fibromyalgia'. The concept and diagnosis became popular, especially in North America, in the 1970s, after the seminal publications of Hugh Smythe (1972) and Smythe and Moldofsky (1977). It is noticeable that there does not appear to be an early case report as there is for instance for gout, rheumatoid arthritis or certain vasculitides. This may be one reason why we still lack a commonly shared clinical image of the 'typical' case. After Smythe and coworkers, operational definitions and classification criteria were given by Yunus et al, Lautenschlager et al (both in 1989) and Wolfe et al in 1990. The latter received the endorsement of the American College of Rheumatology and are now the most widely used. They identify fibromyalgia as a musculoskeletal disorder with spontaneous widespread pain and exaggerated tenderness as prominent and distinctive features. The other two criteria sets refer to a different concept of fibromyalgia as a 'functional' or 'dysfunctional' disorder. These and other nosological differences pose problems for clinical as well as epidemiological research. They may be of minor importance if it is accepted that any present definition is arbitrary and that a wide range of possible elements are more relevant to research than a uniform concept of a disease called fibromyalgia. PMID- 7497540 TI - Soft tissue syndromes. PMID- 7497543 TI - A short history of non-vascular interventional radiology. PMID- 7497544 TI - [In-situ hybridization in the detection of trisomy 12 in chronic lymphatic leukemia]. PMID- 7497542 TI - [Type B Haemophilus influenzae infections. Experience at the Pediatric Hospital of Luxembourg]. AB - We have made a retrospective study of invasive Hib infections which have occurred in the Children's Hospital since 1980 to the end of July 1994; and we have been able to notice that since the introduction of vaccination against Hib, especially since it is free, the incidence of invasive Hib infections in the Grand Duchy of Luxembourg is in great decline. PMID- 7497545 TI - [Genetics of breast cancer]. PMID- 7497547 TI - [General effects of thermal baths]. PMID- 7497548 TI - [Critical bibliographic analysis of the international medical literature in the domain of thermal research]. PMID- 7497546 TI - [European thermal baths and the role of Mondorf-les-Bains in this context. The therapeutic role of thermal baths at the dawn of the year 2000]. PMID- 7497549 TI - [Lingual attachments therapy: another orthodontic approach]. PMID- 7497551 TI - [The role of Mondorf in the European context]. PMID- 7497550 TI - [Comparative status of thermal baths in various European countries]. PMID- 7497552 TI - The clinical utility of epidural midazolam for inguinal hernia repair in children. PMID- 7497554 TI - Lateral popliteal sciatic nerve block compared with ankle block for analgesia following foot surgery. AB - The purpose of this study was to compare postoperative analgesia after foot surgery in patients anaesthetised either with popliteal sciatic nerve block or ankle block. In a prospective study, 40 patients scheduled for foot surgery involving osteotomies were allocated randomly into one of two groups. Following induction of general anaesthesia, Group PS (n = 21) received a lateral popliteal sciatic nerve block and Group AB (n = 19) received an ankle block. Both groups received 20 ml bupivacaine 0.5% plain. In group PS 43% required morphine analgesia in the Post Anaesthetic Recovery Room (PAR) compared with 16% in group AB. Postoperative analgesia was assessed using VAS and a pain scale in the PAR and a questionnaire by telephone follow-up on the day after surgery. The ankle block appeared to be more reliable, providing more consistent analgesia in the PAR. Postoperative analgesia in Group PS lasted a median of 18.0 hr and in Group AB lasted 11.5 hr (P < 0.05). Both techniques provided effective analgesia after discharge home and high levels of patient satisfaction. PMID- 7497553 TI - Midazolam for caudal analgesia in children: comparison with caudal bupivacaine. AB - In a randomized, double-blind study we have examined the analgesic efficacy of caudal administration of midazolam, bupivacaine, or a mixture of both drugs in 45 children, undergoing inguinal herniotomy. They were allocated randomly into three groups (n = 15 in each) to receive a caudal injection of either 0.25% bupivacaine 1 ml.kg-1 with or without midazolam 50 micrograms.kg-1 or midazolam 50 micrograms.kg-1 with normal saline 1 ml.kg-1. There were no differences in quality of pain relief, postoperative behaviour or analgesic requirements between the midazolam group and the other two groups. Times to first analgesic administration (paracetamol suppositories) were longer (P < 0.001) in the bupivacaine-midazolam group than in the other two groups. Further, the bupivacaine-midazolam group received fewer (P < 0.05) doses of paracetamol than the bupivacaine group. Side effects such as motor weakness, respiratory depression or prolonged sedation were not observed in patients who received caudal epidural midazolam only. We conclude that caudal midazolam in a dose of 50 micrograms.kg-1 provides equivalent analgesia to bupivacaine 0.25%, when administered postoperatively in a volume of 1 ml.kg-1 for children following unilateral inguinal herniotomy. PMID- 7497555 TI - Post-tonsillectomy infiltration with bupivacaine reduces immediate postoperative pain in children. AB - Pain management after tonsillectomy in children remains a dilemma for the anaesthetist. A previous study demonstrated that the administration of lidocaine 1% topical spray to the peritonsillar fossae before tracheal extubation provided considerable immediate postoperative pain relief in infants and children. However, the pain relief was of short duration. We were hopeful that the use of bupivacaine would offer more prolonged pain relief because of its pharmacological characteristics. Therefore, this study was designed to compare the effects of bupivacaine 0.5% with 1:200,000 epinephrine administered after tonsillectomy either as topical spray or submucosal infiltration on postoperative pain in children. Forty-three patients aged two to ten years were randomized into three groups after tonsillectomy was performed. Group (1) received 0.5 ml.kg-1 normal saline spray; (2) received 2 mg.kg-1 bupivacaine 0.5% with 1:200,000 epinephrine peritonsillar infiltration in a similar volume to Group 1 and; (3) received 2 mg.kg-1 bupivacaine 0.5% with 1:200,000 epinephrine spray to both tonsillar beds. The patients in each group were compared postoperatively with regard to the quality of pain control using the Objective Pain Score, and their analgesic requirements. Peritonsillar infiltration of bupivacaine provided superior immediate postoperative analgesia as reflected by lower recovery room pain scores (P < 0.05) and opioid requirements (P < 0.01). Ward pain scores and analgesic requirements were similar among groups. Peritonsillar infiltration of bupivacaine 0.5% with 1:200,000 epinephrine provides better post-tonsillectomy pain control in the immediate postoperative period than bupivacaine spray or placebo. PMID- 7497556 TI - A comparison of percutaneous and operative tracheostomies in intensive care patients. AB - The aim of our study was to compare the complication rate of convenional surgical and percutaneous dilational tracheostomies performed under general anaesthesia in critically ill patients. Fifty-three consecutive patients whose lungs were mechanically ventilated and who required tracheostomy were randomised to undergo either conventional surgical tracheostomy (n = 28) in the operating room or percutaneous dilational tracheostomy (n = 25) in the intensive care unit under general anaesthesia. All of the procedures were successfully completed. No deaths were related to the performance of either tracheostomy technique. Three patients in each group required a dressing change for minor bleeding at the tracheostomy site. There was no major bleeding requiring blood transfusion. One patient in each group developed atelectasis detected on chest x-ray post-operatively. In the surgical tracheostomy group, there were two patients with cuff leaks, one with a stomal infection and one with a pneumothorax. None of these complications occurred after percutaneous, dilational tracheostomy. We conclude that the low incidence of complications in both groups indicates that percutaneous dilational tracheostomy can be performed as safely in the intensive care unit with general anaesthesia as surgical tracheostomy can be performed in the operating room. PMID- 7497558 TI - Effects of calcium channel blockers on circulatory response to tracheal intubation in hypertensive patients: nicardipine versus diltiazem. AB - We studied the circulatory responses to laryngoscopy and tracheal intubation in 37 hypertensive patients who received nicardipine 30 micrograms.kg-1 iv (Group N, n = 12), diltiazem 0.3 mg.kg-1 (Group D, n = 12) or saline placebo (Group C, n = 13) 60 sec before the initiation of laryngoscopy. Anaesthesia was induced with thiopentone 5 mg.kg-1 iv, and succinylcholine 2 mg.kg-1 iv was used to facilitate tracheal intubation after precurarization with vecuronium 0.02 mg.kg-1 iv. In patients in Group C heart rate (HR) increased from 79 +/- 14 (baseline) to 110 +/ 12 (P < 0.05) associated with tracheal intubation; mean arterial pressure (MAP) increased from 116 +/- 8 to 140 +/- 77 (P < 0.05) and rate-pressure product (RPP) increased from 13385 +/- 2393 to 21251 +/- 3883 (P < 0.05). The changes from baseline values in HR and RPP after tracheal intubation in Group D were less than those in Groups C and N (P < 0.05). The increase in MAP following tracheal intubation in Groups N and D was lower than that in Group C (P < 0.05). We conclude that, compared with nicardipine, administration of diltiazem iv is associated with less circulatory response to tracheal intubation in hypertensive patients. PMID- 7497557 TI - Speed and ease of tracheal intubation: priming with mivacurium compared with succinylcholine. AB - This study examined the efficacy of mivacurium priming (0.015 mg.kg-1) with five minutes between the priming and intubating doses by comparing the effects of one, two and three times the ED95 dose (0.075 mg.kg-1) of mivacurium after priming (Groups 1, 2 and 3, respectively), with a saline prime and 2 x ED95 mivacurium (Group 4) or 1 mg.kg-1 dose of succinylcholine (Group 5). The time from the intubating dose injection to intubation was measured and intubating conditions were rated on a five-point scale with 4 being optimal and 0 being failure. Mean times (+/- SEM) in seconds between the administration of the intubating dose and tracheal intubation were: 106.4 +/- 5.1, 89.6 +/- 6.7, 81.9 +/- 2.7, 169.9 +/- 7.8 and 82.9 +/- 3.5 for Groups 1-5 respectively. The times for Group 2 (2 x ED95 with priming), Group 3 (3 x ED95 with priming) and Group 5 (succinylcholine with saline) were shorter than the times of Groups 1 (1 x ED95 with priming) and 4 (2 x ED95 with saline) P < 0.05. Mean intubating condition scores (+/- SD) for the five groups respectively were 3.1 +/- 0.6, 3.4 +/- 0.6, 3.5 +/- 0.5, 3.2 +/- 0.6 and 3.8 +/- 0.4. Scores for Groups 2, 3 and 5 were higher than those of Group 1 (P < 0.05). The data demonstrated that (1) priming with mivacurium shortens the intubation time and is accompanied by good intubating conditions with doses 2x and 3x ED95, and (2) intubating times and conditions similar to those achieved with succinylcholine can be obtained using mivacurium 2x (total dose 0.150 mg.kg 1) or 3 x ED95 (total dose 0.215 mg.kg-1) with a five-minute priming interval. Priming provides an alternative technique in those clinical circumstances where succinylcholine is contraindicated. PMID- 7497559 TI - Perioperative administration of caffeine tablets for prevention of postoperative headaches. AB - Interruption of daily caffeine consumption can cause caffeine withdrawal headache. As headache ranks among the most frequent minor postoperative sequelae, the impact of perioperative substitution of caffeine on the incidence of postoperative headache was evaluated. Forty patients undergoing minor surgical procedures with general anaesthesia were randomly allocated to receive either placebo or caffeine tablets at a dosage equal to their individual average daily caffeine consumption. Daily dietary intake was calculated based on an average week-day consumption using conversion factor from previously published sources. The patients were instructed at the preoperative visit to abstain from all external sources of caffeine. Compliance with these dietary restrictions was verified by blood samples obtained immediately before the surgical procedure and on postoperative day 1. The patients were assessed for headache using a standardised checklist immediately before induction of anaesthesia, on the evening of the day of surgery and on the morning of postoperative day 1. Ten patients (50%) who received placebo reported headaches, which persisted in seven patients (35%) until the next day. No patient receiving caffeine substitution therapy reported headache following surgery, and only one complained of headache on postoperative day 1. We suggest that the prophylactic administration of caffeine tablets might be considered for surgical patients who are accustomed to a high daily intake of caffeine. PMID- 7497560 TI - Ranitidine suspension or famotidine resoriblette and gastric fluid volume and pH. AB - We studied the effect of two new formulations of H2-receptor antagonists on gastric fluid pH and volume. Forty-five healthy, elective adult in-patients in three study groups, 15 in each, were premedicated using oral diazepam 10 mg with 100 ml of a dose of water soluble suspension of ranitidine 300 mg with sodium citrate/bicarbonate, or a resoriblette of famotidine 40 mg, or placebo. Gastric fluid samples were obtained by blind aspiration after anaesthesia induction, 50 70 min from premedication, and again 90 min from premedication. After a mean period of 60 min from ingestion the patients medicated with H2-antagonists had higher gastric juice pH than those in the control group (1.5 (1.1-6.3), median (range)) (P < 0.0001) for ranitidine (6.8 (4.1-7.8)), P < 0.01 for famotidine (3.9 (1.5-7.6)); P < 0.05 ranitidine vs famotidine). Recovered volumes were similar for the groups (median 3-4 ml, range 0-50 ml). None of the H2 patients had pH < 3.5 and volume > or = 0.3 ml.kg-1 (P < 0.05 vs placebo). In second aspirations, taken 90 min from premedication, the group differences from control in pH persisted. Famotidine patients had the lowest volumes (P < 0.05 vs controls); yet one famotidine patient had a pH < 2.5 and volume > or = 0.3 ml.kg 1. It is concluded that, at the moment of oral anxiolytic premedication, ranitidine-buffer suspension effectively reduced gastric juice acidity, whereas famotidine resoriblette failed to increase reliably gastric pH in 50-90 min. PMID- 7497562 TI - Propofol emulsion and bacterial contamination. AB - Package insert information provided with propofol advises prompt administration following its drawing-up. This study has examined the delays which occur between drawing-up and administration of propofol in clinical practice and the incidence of bacterial contamination occurring under such conditions. Two hundred and fifty four clinical uses of propofol were examined. Mean elapsed times (range) from drawing-up to induction were 28.8 min (1-172), and 11.6 min (1-65) from induction to culture inoculation. The delay to induction exceeded ten minutes in 68.5% of propofol uses. Sixteen cultures (6.3%) grew bacteria. Delay to induction was not associated with increased chances of bacterial growth in any of the samples. Increasing delay between induction and culture inoculation was associated with greater odds of bacterial growth, which is consistent with contamination occurring at or after induction. Whilst the manufacturers advise prompt administration, our findings show that when inadvertent delays occur, propofol remains bacteriologically safe to use under standard clinical conditions. Microbial contamination can occur at any stage, thus attention to asepsis remains important throughout the administration period. PMID- 7497561 TI - A comparison of intravenous ranitidine and omeprazole on gastric volume and pH in women undergoing emergency caesarean section. AB - We have compared the effect of intravenously administered omeprazole and ranitidine on gastric contents in a double-blind study in 80 consecutive women undergoing emergency Caesarean section. When the decision to perform emergency Caesarean section was made, patients were randomly assigned to receive either ranitidine 50 mg or omeprazole 40 mg intravenously. The volume and pH of the gastric contents were measured immediately after tracheal intubation and again before extubation. The gastric pH was found to be higher after omeprazole than after ranitidine immediately after intubation (5.89 +/- 1.46 and 5.21 +/- 1.36 respectively) (P < 0.05) and before extubation (5.97 +/- 1.38 and 5.32 +/- 1.24 respectively) (P < 0.05). However, the gastric volumes were comparable in both the groups. The number of patients with gastric volume > 25 ml and pH < 2.5 were 3 (7.5%) in the ranitidine group and 1 (2.5%) in the omeprazole group after intubation and none in either of the groups before extubation. We conclude that omeprazole 40 mg iv administered at the time of the decision to operate, results in higher gastric pH than ranitidine in obstetric patients undergoing emergency Caesarean section. PMID- 7497563 TI - Temporary back and leg pain after bupivacaine and morphine spinal anaesthesia. AB - Transient neurological symptoms have been reported after hyperbaric lidocaine 5% spinal anaesthetics. We report a patient with neurogenic back and leg pain after uncomplicated bupivacaine and morphine spinal anaesthesia. A healthy 39-yr-old woman received 1.6 ml hyperbaric bupivacaine 0.75% and 250 micrograms morphine intrathecally. Two hours later, the patient experienced discomfort during suturing of the peritoneum and surgery was completed under general anaesthesia. Recovery was uncomplicated until 13 hr after intrathecal injection, when the patient complained of burning pain in her back extending to the front of the abdomen and similar pain in her thighs. Neurological consultation was obtained. Treatment was started with amitriptyline and the symptoms resolved slowly. Complete recovery occurred over three months. Further studies to assess symptoms after spinal anaesthesia are indicated. PMID- 7497564 TI - Anaesthetic management of a child with Bartter's syndrome. AB - We report the anaesthetic management of an eight-year-old asthmatic boy with Bartter's syndrome who had bilateral orchidopexy with caudal epidural analgesia. Bartter's syndrome is a rare congenital disorder characterized by hypokalaemic hypochloraemic metabolic alkalosis, hyperaldosteronism, hyperreninaemia and hyperplasia of the juxtaglomerular apparatus of the kidneys. Characteristically, although these patients are normotensive they may be hypovolaemic. They may have unstable baroreceptor responses and show marked resistance to vasopressors. Hence, fluid, acid-base and electrolyte imbalances along with haemodynamic instability pose particular problems in their anaesthetic management. Previous case reports have described the management of these patients with general anaesthesia, our patient had his orchidopexy with caudal epidural analgesia using plain bupivacaine 0.5%. The patient was haemodynamically stable throughout surgery and was comfortable with caudal analgesia as the sole anaesthetic. Hypovalaemia, acid-base status and electrolyte imbalance were treated before instituting caudal epidural analgesia. We present this case report which describes the anaesthetic considerations in the light of the pathophysiology of Bartter's syndrome. PMID- 7497565 TI - Anaesthesia for thyroplasty. AB - The purpose of this report is to describe the anaesthetic considerations for layngoplastic procedures. Thyroplasty is a procedure which restores the voice in unilateral vocal cord paralysis. The procedure employs an external approach via a window cut at the appropriate level in the thyroid ala. A wedge of silastic is inserted against the inner perichondrium, thereby displacing the vocal cord medially and permitting voice production. Correct placement of the implant is assessed by asking the patient to phonate; patient cooperation is therefore necessary at certain times during the procedure. We describe our management of a patient undergoing thyroplasty. The use of a benzodiazepine agonist-antagonist combination provided both optimal operating conditions and patient comfort. PMID- 7497566 TI - Tracheal rupture following blunt chest trauma presenting as endotracheal tube obstruction. AB - In this report, we describe a patient in whom a tracheal tear followed blunt thoracic trauma. The diagnosis was made late resulting in problems with ventilation, endotracheal tube obstruction and cardiac arrest. Difficulties with early recognition of tracheobronchial injuries may be caused by non-specific findings as well as the lack of exposure of physicians to patients with these injuries. The signs and symptoms of tracheobronchial injuries are described, as well as their differential diagnoses. A review of airway management has been made as it requires combined anaesthetic and surgical expertise. Injuries of the trachea may have severe, life-threatening consequences and early diagnosis and management reduce morbidity and mortality. PMID- 7497568 TI - Lightwand intubation: II--Clinical trial of a new lightwand for tracheal intubation in patients with difficult airways. AB - Lightwands have been used to assist in the tracheal intubation of patients with difficult airways for many years. A new lightwand (Trachlight) with a brighter light source and a flexible stylet permits both oral and nasal intubation under ambient light. This study reports the effectiveness of the Trachlight in tracheal intubation in patients with difficult airways. Two groups of patients were studied: Group 1--patients with a documented history of difficult intubation or anticipated difficult airways; Group 2--anaesthetized patients with an unanticipated failed laryngoscopic intubation. In Group 1, the tracheas were intubated using the Trachlight with patients either awake or under general anaesthesia. In Group 2, tracheas were intubated under general anaesthesia using the Trachlight. The time-to-intubation, number of attempts, failures, and complications during intubation for all patients were recorded. Two hundred and sixty-five patients were studied with 206 patients in Group 1, and 59 in Group 2. In most patients, the tracheas were intubated orally (183 versus 23 nasal) during general anaesthesia (202 versus 4 awake) in Group 1. Intubation was successful in all but two of the patients with a mean (+/- SD) time-to-intubation of 25.7 +/- 20.1 sec (range 4 to 120 sec). The tracheas of these two patients were intubated successfully using a fibreoptic bronchoscope. Orotracheal intubation was successful in all patients in Group 2 using the Trachlight with a mean (+/- SD) time-to-intubation of 19.7 +/- 13.5 sec. Apart from minor mucosal bleeding (mostly from nasal intubation), no serious complications were observed in any of the study patients.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7497567 TI - Lightwand intubation: I--a new lightwand device. AB - Tracheal intubation under direct vision using a laryngoscope can be challenging and difficult even in experienced hands. Transillumination of the soft tissues of the neck using a lighted-stylet (lightwand) is one of many effective alternative intubating techniques developed during the past several decades. While many versions of lightwand have been available, each has its limitations. A newly developed lightwand (Trachlight) has incorporated many design modifications. It has a brighter bulb, permitting intubation under ambient light in most cases. A retractable stiff wire within the wand adds flexibility to facilitate both oral and nasal intubation. The flexible wand also allows visual (transillumination) guide to proper placement of the tip of the tracheal tube in the trachea. The Trachlight does have some limitations. It is a light-guided technique in which there is no direct visualization of the upper airway structures. It should be avoided in patients with known anatomical abnormalities of the upper airway and used with caution in patients in whom transillumination of the anterior neck may not be achieved adequately. As with any intubating technique, successful intubation using the Trachlight relies on the preparation of the patient and the operator's skill and experience. PMID- 7497569 TI - Positive end-expiratory pressure and lung compliance: effect on delivered tidal volume. AB - The effects of positive end-expiratory pressure (PEEP) and lung compliance (CL) on delivered tidal volume (VTdel) and ventilator output were evaluated in the following anaesthesia machine/ventilator systems: Narkomed III with a Model AV-E ventilator (III/AV-E system) and an Ohmeda Modulus II with either a 7810 anaesthesia ventilator (II/7810 system) or a Model 7000 anaesthesia ventilator (II/7000 system). With a standard circle anaesthesia breathing circuit connected to a test lung simulating CL, gas flow was measured and integrated over time at each combination of VT settings (VTset), 500 ml or 1000 ml; CL settings, 0.15 to 0.01 L.cm H2O-1 decreased incrementally; and PEEP settings, 0 to 30 cm H2O increased in 5-cm H2O increments. The integral of gas flow at the Y-piece of the breathing circuit was recorded as VTdel and at the output of the ventilator bellows as ventilator output. As CL decreased to 0.01 L.cm H2O-1 and PEEP increased to 30 cm H2O, at VTset of 500 ml and 1000 ml, respective VTdel decreased linearly to 251 +/- 6 ml and 542 +/- 7 with the III/AV-E, 201 +/- 5 and 439 +/- 5, with the II/7810, and 181 +/- 4 and 433 +/- 7 ml with the II/7000 (P < 0.05 among the three systems). Loss in VTdel due to PEEP alone, which increased only slightly when VTset was increased, accounted for an increasingly greater percentage of VTset as it was decreased, which was less pronounced with low CL.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7497570 TI - Halogenated gas embolism prevention by Intralipid. PMID- 7497571 TI - Phaeochromocytoma and calcium channel block. PMID- 7497572 TI - Peribulbar anaesthesia. PMID- 7497573 TI - Filling Tec 5 vaporizers. PMID- 7497574 TI - Fibreoptic intubation. PMID- 7497575 TI - Adaptation of ICU ventilator to deliver isoflurane. PMID- 7497577 TI - Pharmacokinetics and antitumor activity of a new platinum compound, cis malonato[(4R,5R)-4,5-bis(aminomethyl)-2-isopropyl-1, 3- dioxolane]platinum(II), as determined by ex vivo pharmacodynamics. AB - The pharmacokinetics and ex vivo pharmacodynamics studies on cis-malonato[(4R,5R) 4,5-bis(aminomethyl)-2-isopropyl-1, 3- dioxolane]platinum(II) (SKI 2053R, NSC D644591), cisplatin (CDDP), and carboplatin (CBDCA) were performed in beagle dogs. Equitoxic doses of SKI 2053R, CDDP, and CBDCA (7.5, 2.5, and 15.0 mg/kg, respectively) were given by i.v. bolus to three beagle dogs in a randomized crossover study. Plasma samples were analyzed for platinum by flameless atomic absorption spectrophotometry. Plasma concentrations of total and ultrafiltrable platinum for the three drugs declined in a biexponential fashion. The mean area under the concentration-time curve (AUC0-->infinity) determined for ultrafiltrable platinum derived from SKI 2053R, as an active component, was 7.72 +/- 2.74 micrograms h ml-1 (mean +/- SD), with an initial half-life of 0.37 +/- 0.20 h, a terminal half-life of 2.19 +/- 0.93 h, a total clearance of 16.83 +/- 4.76 ml min-1 kg-1, and a steady-state volume of distribution of 1.57 +/- 0.30 l/kg. The ex vivo antitumor activity of SKI 2053R was assessed using the ultrafiltrable plasma against two human lung-adenocarcinoma cell lines (PC-9 and PC-14) and five stomach-adenocarcinoma cell lines (MKN-45, KATO III, SNU-1, SNU 5, and SNU-16) by tetrazolium-dye (MTT) assay and was compared with that of CDDP and CBDCA using an antitumor index (ATI) determined from the ex vivo pharmacodynamic results of inhibition rates (%) versus time curves. The mean ATI value was shown to be ranked in the following order: SKI 2053R > CBDCA > CDDP. The mean ATI values recorded for SKI 2053R and CBDCA were significantly (P < 0.05) higher than that noted for CDDP; however, no statistically significant difference was observed between SKI 2053R and CBDCA, suggesting that the antitumor activity of SKI 2053R is superior to that of CDDP and is equivalent to that of CBDCA. These results suggest that SKI 2053R is a promising candidate for further development as a clinically useful anticancer drug. PMID- 7497576 TI - Patients prefer scented face masks. PMID- 7497578 TI - Accumulation of anthracenyl-amino acid topoisomerase I and II inhibitors in drug sensitive and drug-resistant human ovarian cancer cell lines determined by high performance liquid chromatography. AB - Anthracenyl amino acid/dipeptide conjugates (AADC) represent novel structures rationally designed for their DNA-binding properties. A high-performance liquid chromatography method is described for simultaneous determination of five compounds that exhibit novel mechanisms of action as topoisomerase I and II inhibitors. The method uses an Apex ODS-2 column and a mobile phase of 0.25 M ammonium acetate/trifluoroacetic acid (pH 3) in methanol with gradient elution. Selective detection is achieved by monitoring at 545 nm, with limits of detection ranging between 2 and 4 ng on the column. AADC are recovered from cell sonicates by solid-phase extraction using C2 cartridges, with extraction efficiencies ranging from 84% to 95%. Drug uptake studies were performed with three active compounds in the human ovarian cancer cell line A2780 and its multi-drug resistant counterpart 2780AD. Marked differences were observed in the pattern of cellular accumulation produced by each compound. NU/ICRF 505 (tyrosine derivative) was taken up most avidly, reaching plateau levels of 4000 pmol/10(6) cells after 2 h, with no difference being apparent between A2780 and 2780AD. NU/ICRF 510 (arginine derivative) accumulated slowly in A2780, failing to achieve an equilibrium after 4 h, and appeared to be completely excluded from 2780AD. NU/ICRF 500 (serine derivative) was most rapidly taken up by A2780, producing a plateau of 800 pmol/10(6) cells after only 30 min with approximately 3-fold less accumulation in 2780AD. These results are correlated to the chemosensitivity of the two cell lines to the three compounds. PMID- 7497580 TI - Stability of the i.v. and oral formulations of etoposide in solution. AB - Etoposide is a widely used cytotoxic drug that requires complex formulation for both the i.v. and oral preparation to ensure drug stability. Data on the stability of the i.v. formulation when diluted in infusion fluids are contradictory, and there is little information on the stability of the oral preparation in gastric or intestinal fluids. The stability of both i.v. and oral etoposide was therefore evaluated in the present investigation. The stability of the i.v. preparation was investigated across a range of concentrations in infusion fluids, being determined by regular sampling for high-performance liquid chromatography (HPLC) analysis and by visual inspection. The stability of the oral preparation was studied in both artificial gastric and intestinal fluids, again with regular sampling for HPLC analysis, and the influence of pH, concentration and the addition of ethanol and bile salts on oral stability was determined. The i.v. preparation showed a marked decrease in stability with increasing drug concentration, but stability was additionally reduced in i.v. bags regularly sampled with a syringe and needle as compared with bags that were inspected visually only (minimal stability in sampled bags, 24 h at 0.5 mg/ml and 5 h at 1.0 mg/ml, as compared with 10 days and 18 h at the respective concentrations in unsampled bags). Stability was also greater at room temperature, 20-23 degrees C, as compared with 8-12 degrees C. Loss of stability was indicated by a decrease in etoposide concentration (measured by HPLC) and the appearance of a fine white precipitate, shown to be pure etoposide. Importantly, the appearance of precipitate was as sensitive as a specific HPLC assay in detecting loss of stability and was in many cases apparent when the etoposide concentration was within 5% of the starting concentration. The oral formulation also showed a marked concentration-dependent decrease in stability in artificial intestinal fluid at pH 7.5 (percentage of etoposide in solution after 2 h at 0.5, 1.0, 1.5 and 2.0 mg/ml, 94 +/- 2%, 80 +/- 5%, 68 +/- 13% and 41 +/- 9%, respectively). There was no concentration effect on stability in gastric fluid at pH 3.0, although stability was much greater at pH 3 and pH 5 as compared with pH 1 or in intestinal fluid at pH 7.5.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7497579 TI - Evaluation of plasma 5-fluorouracil nucleoside levels in patients with metastatic breast cancer: relationships with toxicities. AB - This paper describes the relationship between 5-fluorouracil (FUra)-derived toxicities and plasma levels of the FUra anabolites 5-fluorouridine (FUrd) and 5 fluoro-2'-deoxyuridine (FdUrd) monitored in patients receiving continuous infusions of FUra (1000 mg/m2 per 24 h) over 5 days preceded by the administration of cisplatin (100 mg/m2). A total of 63 courses of this treatment were given as second-line chemotherapy to 17 patients with metastatic breast cancer. The active FUra anabolites FUrd and FdUrd were monitored twice daily in the plasma by high-performance liquid chromatography. Data were analyzed using multiple analysis of variance (ANOVA). Only a low proportion of patients exhibited measurable plasmatic levels of FUrd (43%) and FdUrd (70%). The areas under the plasma concentration-time curves (AUC) determined over 120 h for FUrd (AUCFUrd) and for FdUrd (AUCFdUrd) were found to be statistically significantly different for chemotherapy cycles with and those without myelosuppression. Chemotherapy cycles without neutropenia were associated with low AUCFUrd values (mean +/- SEM, 2.9 +/- 0.7 micrograms ml-1 h) and high AUCFdUrd values (14.1 +/- 2.7 micrograms ml-1 h), respectively, whereas courses with myelosuppression (WHO grades 2-4) showed inverse profiles with high AUCFUrd values (16.3 +/- 2.3 micrograms ml-1 h) and low AUCFdUrd values (3.1 +/- 1.0 micrograms ml-1 h), respectively. A statistically significant difference in AUCFdUrd values was also observed between cycles with and those without mucositis (P = 0.0027), with AUCFdUrd values being 22.6 +/- 5.6 and 7.8 +/- 1.9 micrograms ml-1 h, respectively. Whereas hematotoxicity could be correlated with both AUCFUrd and AUCFdUrd values, mucositis was associated with high AUCFdUrd levels. Moreover, a negative correlation was found between the AUCs determined for FUrd and FdUrd (P = 0.002), indicating that activation of FUra via FUrd or via FdUrd may involve competitive processes. Therefore, to follow the development of the major FUra derived toxicities, measurement of FUrd and FdUrd plasma levels appeared very attractive. PMID- 7497582 TI - An open study to assess the safety, tolerance and pharmacokinetics of an intravenous infusion of granisetron given at 3 mg over 30 s in patients receiving chemotherapy for malignant disease. AB - Granisetron is a highly potent and selective 5-hydroxytryptamine3 (5-HT3) receptor antagonist indicated for the prevention of cytotoxic-induced nausea and vomiting. Clinical trials have demonstrated granisetron to be effective and well tolerated at a standard dose of 40 micrograms/kg or 3 mg given i.v. as a 5-min infusion. In this study, the efficacy and safety of granisetron given as a 30-s infusion was assessed. A total of 21 patients, scheduled to undergo chemotherapy, received a single 3-mg i.v. dose of granisetron over 30 s, completed at 1 h before chemotherapy administration. Patients were allowed two further i.v. doses of granisetron at 3 mg within the 24-h assessment period. Changes from baseline values in vital signs were analysed prior to granisetron administration and at 30 s as well as 1, 10, 15, 30 and 60 min after granisetron administration. Holter ECG recordings were taken for 6 h prior to and 1 h after administration. No significant change was found in vital signs at 30 s or 1 min after granisetron infusion. There was a small but statistically significant fall in diastolic blood pressure as compared with baseline and a non-significant trend in favour of a reduction in heart rate at 10 and 15 min. No ECG abnormality was recorded post infusion that had not been present pre-infusion. None of these changes was considered to be clinically relevant. The treatment was well tolerated. The most frequently reported adverse events were constipation (n = 6) and headache (n = 5). Maximal plasma levels of granisetron were within the range of 44.57-410 ng/ml except in one patient. The median values recorded for peak concentration (Cmax) and area under the curve (AUC) were 195 ng/ml and 71.2 ng h ml-1, respectively. In conclusion, granisetron at 3 mg was shown to be safe and well tolerated when given as a 30-s i.v. infusion to patients receiving chemotherapy for malignant disease. PMID- 7497581 TI - Pharmacological attempts to improve the bioavailability of oral etoposide. AB - Etoposide demonstrates incomplete and variable bioavailability after oral dosing, which may be due to its concentration and pH-dependent stability in artificial gastric and intestinal fluids. The use of agents that may influence etoposide stability and, thereby, bioavailability, was investigated in a number of clinical studies. Drugs that influence the rate of gastric emptying, while modulating the time of drug absorption, did not significantly alter the etoposide area under the concentration-time curve (AUC) or bioavailability. Specifically, metoclopramide had little effect on the etoposide absorption profile and did not significantly alter the AUC (AUC with etoposide alone, 68.4 +/- 20.3 micrograms ml-1 h, versus 74.3 +/- 25.9 micrograms ml-1 h with metoclopramide), suggesting that in most patients the drug is already emptied rapidly from the stomach. In contrast, propantheline produced a dramatic effect on etoposide absorption, delaying the time of maximal concentration tmax from 1.1 to 3.5 h (P < 0.01), but again without a significant improvement in drug AUC or bioavailability across the 24-h study period (AUC with etoposide alone 78.3 +/- 19.1 micrograms ml-1 h, versus 88.1 +/- 23.6 micrograms ml-1 h with propantheline). The effect of these drugs on the absorption of oral paracetamol, a drug included in the study as a marker of gastric emptying, was exactly the same as that found for etoposide, with no change in AUC being observed after metoclopramide or propantheline administration but a significant delay in tmax being seen on co-administration with etoposide and propantheline. The co-administration of ethanol or bile salts (agents that significantly improved the stability of etoposide in artificial intestinal fluid) with oral etoposide similarly had no effect on improving the etoposide AUC or reducing the variability in AUC, suggesting that drug stability in vivo was not affected by these agents. In the third study the co-administration of cimetidine had no effect on the pharmacokinetics of oral or i.v. etoposide, despite the previous observation that etoposide stability was markedly improved at pH 3-5 as compared with pH 1 in artificial gastric fluid. This series of studies, designed to investigate factors that improved etoposide stability in laboratory studies, failed to demonstrate any potentially useful improvement in AUC or bioavailability in the clinical setting. PMID- 7497583 TI - Phase I trial of chloroguinoxaline sulfonamide, with correlation of its pharmacokinetics and pharmacodynamics. AB - To define a maximum tolerable dose, chloroquinoxaline sulfonamide (CQS) was given as a 1-h infusion every 28 days to cancer patients for whom no effective standard therapy was available. Doses were escalated in cohorts of at least three patients each. Plasma for characterization of the pharmacokinetics of free and total CQS was obtained during and after the initial infusion and, when possible, during and after subsequent infusions of CQS if the dose had been reduced. A total of 101 courses of CQS in 55 patients were evaluated. Dose levels ranged from 18 to 3,700 mg/m2. The dose-limiting toxicity was hypoglycemia, first recognized at the 3,700 mg/m2 dose. When dose-limiting hypoglycemia was recognized, patients were entered at successively lower doses, with close monitoring of plasma glucose and insulin concentrations being done in 26 patients. Grade 1-3 hypoglycemia occurred within 4 h of the termination of CQS infusion and cleared by 24 h. Symptomatic hypoglycemia was more frequent at doses of CQS above 1,000 mg/m2. Concomitant administration of 5% glucose did not ameliorate the hypoglycemia associated with CQS doses of > 1,000 mg/m2. The total calorie intake, percentage of ideal body weight, or percentage of weight lost did not explain the incidence or severity of hypoglycemia in 12 patients in whom these data were obtained. Cardiac tachyarrhythmias occurred in 7 patients who received CQS at doses of > or = 1,000 mg/m2, and tachyarrhythmia was associated with hypoglycemia in 3 patients. Other toxicities were sporadic, but the frequency of toxicity was higher at CQS doses of > or = 1,000 mg/m2. These toxicities included fever, rash, lightheadedness, leukopenia, thrombocytopenia, alopecia, diarrhea, nausea, and vomiting. All toxicities were reversible. Mean peak plasma [CQS] and AUC increased with dose, with a suggestion that peak plasma [CQS] plateaued at higher doses. The decline in plasma [CQS] was fitted to a three-compartment, open linear model. The terminal half-life ranged from 28 to 206 h. Total body clearance ranged from 44 to 881 ml/h with no evidence of saturation. Urinary excretion of the parent compound in 24 h averaged < 5%. CQS not bound to plasma protein (free CQS) comprised 1%-17% of total plasma CQS and was not related to dose. A relationship was defined between the magnitude of hypoglycemia and CQS pharmacokinetic parameters. The percentage of decrease in plasma [glucose], i.e., (predose [glucose]-nadir [glucose]/predose [glucose]) x 100, correlated with both free and total peak plasma [CQS].(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 7497584 TI - Effects of 5-benzylacyclouridine, an inhibitor of uridine phosphorylase, on the pharmacokinetics of uridine in rhesus monkeys: implications for chemotherapy. AB - The effects of subcutaneous administration of 5-benzylacyclouridine (BAU), a uridine phosphorylase (UrdPase, EC 2.4.2.3) inhibitor, on uridine concentration in plasma and urine were evaluated in rhesus monkeys. Administration of BAU at 50, 100 and 250 mg/kg increased the plasma uridine baseline concentration 1.5-, 2.9-, and 3.2-fold, respectively. The basis for this moderate perturbation of plasma uridine by BAU was investigated using a tracer dose of 500 microCi 3H uridine. Administration of 3H-uridine alone led to its rapid catabolism to uracil and dihydrouracil. Administration of 83.3 mg/kg BAU with 500 microCi 3H-uridine resulted in a 2.5-fold enhancement of 3H-uridine plasma levels and a substantial decrease in the plasma levels of uridine catabolites, suggesting inhibition of UrdPase activity by BAU in rhesus monkeys. Coadministration of 83.3 mg/kg BAU with 83.3 mg/kg uridine also reduced the plasma concentration of uracil and dihydrouracil, but it did not increase plasma uridine concentration above that of control animals receiving 83.3 mg/kg uridine alone. In animals receiving uridine alone at 83.3 or 25 mg/kg, approximately 10% of the administered dose was recovered in the urine within 6 h, with unchanged uridine being the major component. In contrast, administration of 83.3 mg/kg BAU increased the excretion of unchanged uridine to more than 32% of the total dose administered, even when the urinary excretion ratio of uracil to uridine was reduced ten-fold. Administration of multiple doses (three times per day) of BAU alone (83.3 mg/kg) or in the presence of uridine (83.3 mg/kg) did not enhance plasma uridine concentration further. In addition, uridine pharmacokinetics were associated with a time-dependent relationship as evidenced by an increased total plasma clearance, renal clearance and volume of distribution, resulting in a substantial decrease in uridine peak concentration with time. These results indicate that administration of BAU inhibits UrdPase activity in rhesus monkeys as manifested by decreased uracil and dihydrouracil plasma levels, as well as a lower urinary excretion ratio of uracil to uridine, as compared to control animals. However, plasma levels of unchanged uridine were not substantially enhanced by BAU in spite of the large increase in urinary excretion of unchanged uridine. This phenomenon was also observed when uridine was coadministered with BAU, suggesting that plasma uridine concentration in monkeys may be strongly regulated by the renal system as evidenced by the "spillover" of excess plasma uridine into urine. In addition, the pharmacokinetics of uridine were dose-independent, but time dependent.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 7497585 TI - Selective intra-arterial infusion of high-dose cisplatin in patients with advanced head and neck cancer results in high tumor platinum concentrations and cisplatin-DNA adduct formation. AB - A group of 23 patients with advanced head and neck cancer were treated with highly selective intra-arterial (IA) cisplatin 150 mg/m2 delivered rapidly through microcatheters. The systemic effects of cisplatin were neutralized by concurrent administration of sodium thiosulfate. Two-to-threefold higher tumor platinum contents were detected in tumor biopsies after selective IA cisplatin administration compared to historical controls (treated with 100 mg/m2 IA). Cisplatin-induced DNA modification in human tumor biopsies was quantitated using the antiserum NKI-A59. High levels of cisplatin DNA adducts were detected which correlated linearly with the tumor platinum content (r2 = 0.62). The addition of radiotherapy to this high dose intensity cisplatin treatment resulted in a 92% complete response (CR) rate (12 of 13 patients achieved a CR). Since no difference in tumor platinum content was detected between patients receiving or not receiving radiotherapy (13 and 10 patients, respectively), but the response rate was substantially different (12 CR and 1 partial response with radiotherapy versus 6 partial and 4 non-responders without radiotherapy), these data suggest that the high platinum levels achieved by selective IA infusion were sufficient to produce enough interaction with radiotherapy to cause a 92% CR rate. Whether this interaction is additive or synergistic is as yet unclear. PMID- 7497586 TI - Dosing of thioTEPA for myeloablative therapy. AB - High-dose thioTEPA is used frequently in myeloablative regimens for marrow transplantation, but the need for dose adjustments in obese patients has not been explored. We determined the pharmacokinetics of thioTEPA and its metabolite TEPA during first-dose infusion of thioTEPA 150-250 mg/m2 given daily for 3 days in combination with busulfan and cyclophosphamide, and evaluated the results for correlations with toxicity and dosing strategies. The study included 15 adults undergoing marrow transplantation for hematologic malignancies. Plasma samples were obtained at various times over a 24-h period, and concentrations of thioTEPA and TEPA were measured by gas chromatography. At 22-24 h after initiation of a 4 h infusion, the mean +/- SE plasma concentration of thioTEPA was 124 +/- 63 ng/ml, while that of TEPA was 235 +/- 69 ng/ml. For CFU-GM and BFU-E growth in vitro, the IC50(s) of thioTEPA were 83 ng/ml and 16 ng/ml, respectively, and the IC50(s) of TEPA were 141 ng/ml and 47 ng/ml, respectively. Using a two compartment model, the mean thioTEPA Vc was 47.4 +/- 4.7 1/m2, t1/2 alpha 19 +/- 5 min, t1/2 beta 3.7 +/- 0.5 h, and plasma clearance 302 +/- 21 ml/min per m2. The mean AUCs were 6.9-16.2 mg h/1 for thioTEPA and 8.9-21.2 mg h/1 for TEPA, while the mean peak concentrations were 0.95-2.08 micrograms/ml for thioTEPA and 0.88-1.90 micrograms/ml for TEPA. There was a significant association of grades 2 4 maximum regimen-related toxicity (RRT) with TEPA peak > 1.75 micrograms/ml and with combined thioTEPA and TEPA AUC > 30 mg h/1 (5/6 vs 0/9, P = 0.01 for both comparisons), suggesting that drug exposure was an important determinant of toxicity and, potentially, efficacy. ThioTEPA Vc correlated best with adjusted body weight (r = 0.74, P = 0.0015). In an evaluation of 74 adults receiving thioTEPA 750 mg/m2 in combination with busulfan and cyclophosphamide, the maximum RRT for patients at ideal weight was significantly greater than that for obese patients dosed on ideal weight (mean RRT grade 1.7 vs 1.0, P = 0.004) but did not differ from the maximum RRT for obese adults dosed on actual or adjusted weights. We recommend that for obese patients thioTEPA be dosed on adjusted body weight. Measurements at time-points after 24 h are needed to determine when thioTEPA and TEPA concentrations are below myelosuppressive levels and safe for marrow infusion. PMID- 7497587 TI - Pharmacokinetics and pharmacodynamics of prolonged oral etoposide in women with metastatic breast cancer. AB - The pharmacokinetics and pharmacodynamics of prolonged oral etoposide chemotherapy were investigated in 15 women with metastatic breast cancer who received oral etoposide 100 mg as a single daily dose for up to 15 days. There was considerable interpatient variability in the day 1 pharmacokinetic parameters: area under the plasma concentration time curve (AUC) (0-24 h) 1.95 +/ 0.87 mg/ml per min (mean +/- SD), apparent oral clearance 60.9 +/- 21.7 ml/min per 1.73 m2, peak plasma concentration 5.6 +/- 2.5 micrograms/ml, time to peak concentration 73 +/- 35 min and half-life 220 +/- 83 min. However, intrapatient variability in systemic exposure to etoposide was much less with repeated doses. The intrapatient coefficient of variation (CV) of AUC for day 8 relative to day 1 was 20% and for day 15 relative to day 1 was 15%, compared to the day 1 interpatient CV of 45%. Neutropenia was the principal toxicity. Day 1 pharmacokinetic parameters were related to the percentage decrease in absolute neutrophil count using the sigmoidal Emax equation. A good fit was found between day 1 AUC and neutrophil toxicity (R2 = 0.77). All patients who had a day 1 AUC > 2.0 mg/ml per min had WHO grade III or IV neutropenia. The predictive performance of the models for neutrophil toxicity was better for AUC (percentage mean predictive error 5%, percentage root mean square error 18.1%) than apparent oral clearance, peak plasma concentration, or daily dose (mg/m2). A limited sampling strategy was developed to predict AUC using a linear regression model incorporating a patient effect. Data sets were divided into training and test sets. The AUC could be estimated using a model utilizing plasma etoposide concentration at only two time points, 4 h and 6 h after oral dosing (R2 = 98.9%). The equation AUCpr = -0.376 + 0.631 x C4h + 0.336 x C6h was validated on the test set with a relative mean predictive error of -0.88% and relative root mean square error of 6.4%. These results suggest monitoring of AUC to predict subsequent myelosuppression as a strategy for future trials with oral etoposide. PMID- 7497588 TI - Collateral sensitivity to radiation and cis-platinum in a multidrug-resistant human leukemia cell line. AB - Although collateral sensitivity to gamma radiation has previously been described in multidrug-resistant tumor cell lines, we describe here a multidrug-resistant human T-cell acute lymphatic leukemia cell line, L100, which displayed increased sensitivity to both gamma radiation and cis-platinum. Cis-platinum cytotoxicity of parental L0 cells and L100 cells was enhanced, whereas radiation sensitivity of L0 and L100 cells was unaltered by glutathione depletion. These results indicate that disparate mechanisms are operative in the collateral sensitivity of L100 cells to gamma radiation and cis-platinum. PMID- 7497589 TI - The distribution of systemically administered [3H]-paclitaxel in rats: a quantitative autoradiographic study. AB - Paclitaxel is an important agent in the treatment of many common malignancies. Although the symptomatic peripheral neuropathy caused by this drug is its principal nonhematologic toxicity, little is known about the distribution of paclitaxel within the peripheral or central nervous system following systemic administration. In order to study paclitaxel's distribution in neural and extraneural tissues, adult Sprague-Dawley rats were sacrificed 2 h after a tail vein injection of [3H]-paclitaxel (0.03 mg/kg, 250 microC/rat). Samples of lung, heart, liver, spleen, kidney, skeletal muscle, brain, spinal cord, dorsal root ganglion, and peripheral nerve were then removed and snap-frozen. These tissues were sectioned at 10 microns in a cryostat and exposed to autoradiography film for 2 weeks. The distribution and concentrations of [3H]-paclitaxel in plasma, urine and cerebrospinal fluid were also determined using liquid scintillation spectrometry. [3H]-paclitaxel concentrations (and organ/plasma concentration ratios) in plasma, urine and cerebrospinal fluid were 2.6 nM (1), 38 nM (15) and 0.7 nM (0.3), respectively. A relatively homogeneous distribution of [3H] paclitaxel was observed in liver [412 nM (151)], spleen [351 nM (133)], heart [319 nM (117)], lung [268 nM (93)] and muscle [69 nM (26)]. Higher concentrations of [3H]-paclitaxel were noted in the portal triads [869 nM (361)], glomeruli [797 nM (304)], and renal medulla [961 nM (363)], which may reflect biliary excretion and glomerular filtration. A high concentration of [3H]-paclitaxel was also noted in the choroid plexus [432 nM (167)], but [3H]-paclitaxel was not detected in the brain parenchyma, spinal cord, dorsal root ganglion, peripheral nerve, or the testicles. The pathogenesis of paclitaxel-induced neurotoxicity remains obscure given its limited distribution in the nervous system. In addition, these results suggest that systemically administered paclitaxel is not likely to be effective for the treatment of malignancies in the testes or the nervous system. PMID- 7497590 TI - Morphonuclear characterization of drug resistance by means of digital cell-image analysis: an in vitro assessment. AB - The prediction of tumor resistance to antineoplastic drugs remains an important challenge in cancer chemotherapy. Several methods have been proposed in this connection, but they present a number of problems such as clinical relevance and applicability. In the present work we put forward an original methodology to assess the drug sensitivity of cancer cells. For this purpose we submitted chemosensitive and chemoresistant cell lines to different anticancer drugs and monitored the cell growth and the drug-induced morphonuclear effects by means of digital cell-image analysis of Feulgen-stained nuclei. The results showed that drug-induced effects at the morphonuclear level correlated statistically with the effects produced at the cell proliferation level. For example, the mean nuclear size value increased as a function of the drugs' efficiency recorded at the cell proliferation level. In the same way, the frequency of large dense chromatin clumps also increased in accordance with the drugs' efficiency. The present work thus demonstrates that digital cell-image analysis can be applied to monitor the efficiency of chemotherapeutic treatment carried out on cell lines in vitro. The present methodology could possibly be used on solid tumors, from which biological material can be obtained serially by means of fine-needle aspiration. As evidence of this, the present methodology can also be applied to hematological cancers. PMID- 7497591 TI - Functional expression of the renal organic cation transporter and P-glycoprotein in Xenopus laevis oocytes. AB - The hypothesis that P-glycoprotein (P-gp) mediates the renal secretion of organic cations was tested by functional expression of mRNAs in the Xenopus laevis oocyte system. Efflux of 2'-deoxytubercidin (dTub), a substrate for the renal organic cation transporter (OCT) but not for P-gp, was enhanced by injection of renal mRNA but not by injection of mRNA from P-gp-overexpressing cells (MDCK cells transduced with the cDNA for human MDR1). The functional capacity of the MDCK-MDR mRNA was established by its ability to reduce the steady-state uptake of a classical P-gp substrate, vinblastine. Thus, these data indicate OCT and P-gp to be distinct entities. The Xenopus oocyte system provides a functional approach to further characterize the OCT. PMID- 7497592 TI - High-dose 5-fluorouracil and leucovorin as second-line chemotherapy in patients with platinum-resistant epithelial ovarian cancer. AB - A total of 14 patients with platinum-resistant advanced epithelial ovarian cancer were treated with a continuous infusion of high-dose 5-fluorouracil (5-FU, 1200 mg/m2 per day) for 2 consecutive days weekly for 4 weeks and, thereafter, every 2 weeks in combination with a push injection of folinic acid (20 mg/m2) given just before 5-FU and after 24 h. No objective response was documented, and only five patients showed stable disease. The median survival was 6.5 months. There was minimal toxicity. This schedule of 5-FU in combination with folinic acid is not effective as second-line chemotherapy in advanced ovarian cancer. PMID- 7497594 TI - Intraperitoneal 5-fluoro-2'-deoxyuridine (FUDR) and (S)-leucovorin for disease predominantly confined to the peritoneal cavity: a pharmacokinetic and toxicity study. AB - Intraperitoneal (IP) administration of fluorinated pyrimidines has been evaluated for ovarian and gastrointestinal malignancies in phase I, II, and III trials. The tolerance and pharmacokinetic profile of IP 5-fluoro-2'-deoxyuridine(FUDR) alone and with (R,S)-leucovorin ((R,S)-LV) have each been evaluated in previous phase I studies. FUDR doses of 3 g per day with and without (R,S)-LV doses up to 640 mg per day given IP are well tolerated. The current phase I study was designed to determine the pharmacokinetic profiles and clinical tolerance of escalating doses of the pure biologically active S-isomer of leucovorin ((S)-LV) given IP with the same dosing schedule of FUDR. A group of 16 patients with disease confined to the abdominal cavity were treated in this study. Pharmacokinetic studies of blood and peritoneal fluid, toxicity profiles, and clinical response for the first three cycles are reported here. The toxicity profile did not significantly differ from the prior two studies. All non-hematologic toxicities, such as fatigue, nausea, vomiting, diarrhea, and abdominal discomfort were less than grade 4, and most were less than grade 3. Neutropenia and thrombocytopenia were uncommon and observed only in patients with compromised bone marrow reserve. The pharmacokinetic profiles were also congruent with the previous studies and indicate a three-log advantage for FUDR. The (S)-LV profiles in the peritoneal cavity paralleled those of FUDR. Antitumor effects or absence of progression until after cessation of therapy were documented in 11 patients. At a median follow-up of 18 months 44% of patients were alive. IP administration of 3-g of FUDR and up to 640 mg (S)-LV daily for three days was well tolerated. The tolerance and antitumor effects observed during IP FUDR and LV in these studies encourage further exploration of this regimen against ovarian and gastrointestinal malignancies. The actual role and optimal dose of LV as an enhancer of the antitumor actions of FUDR administered by this route remain unknown. PMID- 7497593 TI - Pharmacokinetics and tissue distribution of cisplatin in nude mice: platinum levels and cisplatin-DNA adducts. AB - The pharmacokinetics of platinum (Pt) and cisplatin (CDDP)-DNA adducts were studied in nude mice after single-dose CDDP treatments. Whole blood, serum, kidney, lever, testis, brain, and tumor were collected at different intervals after injection of CDP at different dose levels. Pt was measured with flameless atomic absorption spectrometry (FAAS) or adsorptive voltammetry (AdV) and CDDP DNA adducts with quantitative immunohistochemistry. The drug was immediately absorbed into the blood circulation (peak serum Pt levels were reached within 5 min) after i.p. CDDP administration, and distribution into most tissues also occurred rapidly (tissue Pt levels peaked at 15 min). With a sampling period of 7 days there was a biphasic elimination of Pt from blood, serum, and tissues. In the brain the pharmacokinetics differed with a gradual accumulation of Pt occurring during the 1st week. Formation of CDDP-DNA adducts in tissues was a slower process, with maximal levels being achieved at between 30 min and 4 h after drug administration, followed by a steady state lasting for at least 24 h. Each tissue type had its specific immunohistochemical staining pattern of adducts. With escalating CDDP doses there was a linear, or almost linear, increase in Pt concentrations and CDDP-DNA adduct levels in all sample types examined. These results suggest that a fair estimation of the amount of drug in tumor and normal tissues can be made from analysis of serum Pt at a fixed time point after a single dose of CDDP. PMID- 7497595 TI - A phase I trial of 5-day continuous infusion cisplatin and interferon alpha. AB - Combination therapy of cisplatin with interferon alpha (IFN) has been shown in several in vitro as well as in vivo models to be synergistic. In order to decrease toxicity seen with cisplatin, 5-day continuous infusions, in place of bolus administration, have been introduced. This led us to investigate the combination of 5-day continuous infusion cisplatin with repeated IFN dosing in a phase I cisplatin dose escalation study. A group of 17 patients were enrolled in this trial. The maximum tolerated dose (MTD) of cisplatin was 20 mg/m2 per day when combined with 3 x 10(6) units IFN given three times a week. The dose limiting toxicities seen included thrombocytopenia, leukopenia, and nausea and vomiting. Pharmacokinetic analyses of free (unbound or ultrafilterable) platinum revealed that the decay curve fitted a monoexponential model. Pharmacokinetic parameters of cisplatin were found to correlate with toxicity. Both increases in the maximum concentration of cisplatin achieved (Cpmax) as well as the area-under the-curve (AUC) for free platinum, correlated with the incidence of nausea and vomiting (both acute and delayed) and hematological toxicities (leukopenia and thrombocytopenia). None of the patients exhibited significant changes in renal function while on this study. The free platinum levels were higher than found in similar studies evaluating comparable cisplatin infusions alone. The enhanced toxicities seen in this trial may be explained by the results of an in vitro study using human plasma spiked with cisplatin and IFN that revealed decreased protein binding of cisplatin by 2.5-3.0-fold. Of the 17 patients treated, two non small cell lung cancer patients obtained a partial response and one malignant melanoma patient obtained complete resolution of a malignant pleural effusion. Considering the acceptable toxicity seen in this trial, we recommend phase II trials be conducted with continuous infusion cisplatin with IFN in the treatment of non-small cell lung cancer. PMID- 7497596 TI - Population pharmacokinetics of docetaxel during phase I studies using nonlinear mixed-effect modeling and nonparametric maximum-likelihood estimation. AB - Docetaxel, a novel anticancer agent, was given to 26 patients by short i.v. infusion (1-2 h) at various dose levels (70-115 mg/m2, the maximum tolerated dose) during 2 phase I studies. Two population analyses, one using NONMEM (nonlinear mixed-effect modeling) and the other using NPML (nonparametric maximum likelihood), were performed sequentially to determine the structural model; estimate the mean population parameters, including clearance (Cl) and interindividual variability; and find influences of demographic covariates on them. Nine covariates were included in the analyses: age, height, weight, body surface area, sex, performance status, presence of liver metastasis, dose level, and type of formulation. A three-compartment model gave the best fit to the data, and the final NONMEM regression model for Cl was Cl = BSA(Theta1 + Theta02 x AGE), expressing Cl (in liters per hour) directly as a function of body surface area. Only these two covariates were considered in the NPML analysis to confirm the results found by NONMEM. Using NONMEM [for a patient with mean AGE (52.3 years) and mean BSA (1.68 m2)] and NPML, docetaxel Cl was estimated to be 35.6 l/h (21.2 lh-1 m-2) and 37.2 l/h with interpatient coefficients of variations (CVs) of 17.4% and 24.8%, respectively. The intraindividual CV was estimated at 23.8% by NONMEM; the corresponding variability was fixed in NPML in an additive Gaussian variance error model with a 20% CV. Discrepancies were found in the mean volume at steady state (Vss; 83.21 for NPML versus 1241 for NONMEM) and in terminal half-lives, notably the mean t1/2 gamma, which was shorter as determined by NPML (7.89 versus 12.2 h), although the interindividual CV was 89.1% and 62.7% for Vss and t1/2 gamma, respectively. However, the NPML-estimated probability density function (pdf) of t1/2 gamma was bimodal (5 and 11.4 h), probably due to the imbalance of the data. Both analyses suggest a similar magnitude of mean Cl decrease with small BSA and advanced age. PMID- 7497598 TI - Antimicrotubule effects of the novel antitumor benzoylphenylurea derivative HO 221. AB - The antitumor action of HO-221, a novel benzoylphenylurea derivative, was studied. The in vitro cytotoxic strength of HO-221 was investigated, as measured by IC50 values, compared with those of other drugs with different action mechanisms, using Chinese hamster lung (CHL) cells, mouse leukemia L1210 cells and human promyelocytic leukemia HL-60 cells. Morphological alterations following treatment were observed under a phase contrast microscope, and the mitotic index was determined at regular intervals to check for accumulation of metaphase cells. HO-221 was found to have a very strong toxic effect on all cell types, equal to that of the spindle poisons used as controls. HO-221 also produced the same specific morphological changes as the spindle poisons, with a significant accumulation of metaphase cells. A chromosome analysis of treated cells showed that HO-221 frequently induced polyploid and aneuploid cells, but without accompanying chromosome-breaking activity. An in vivo mouse bone marrow micronucleus assay was also carried out. The assay allowed the in vivo identification of a chromosome breaker or a spindle poison through the measurement of the relative sizes of micronuclei produced and erythrocytes. HO 221 was found frequently to induce relatively large micronuclei, an action regarded as specific to spindle poisons. It was thus demonstrated that HO-221 acts as a spindle poison both in vitro and in vivo. In order to investigate the mechanism of this action, a study of tubulin assembly using purified calf brain tubulin was carried out, which demonstrated clearly that HO-221 inhibits microtubule assembly. A detailed investigation of the action mechanism of HO-221 as a spindle poison is now called for. PMID- 7497599 TI - Comparative pharmacodynamic analysis of TAT-59 and tamoxifen in rats bearing DMBA induced mammary carcinoma. AB - TAT-59 suppressed the growth of DMBA-induced mammary tumors in rats earlier and more strongly than tamoxifen (TAM). After oral administration of the drugs, DP TAT-59, one of the main metabolites of TAT-59, was found in 10- to 15-fold higher concentrations in both the tumor and blood compared to 4-OH-TAM, an active metabolite of TAM. In a 3-day antiuterotrophic test, every detected metabolite of TAT-59 showed stronger antiestrogenic activity than did TAM. In a competition assay, the affinity of the metabolites for estrogen receptors ranged from that of estradiol to that of TAM. These results suggest that the superior antiestrogenic activity of TAT-59 compared to TAM was either due to its higher penetration into tumor tissue or to the stronger antiestrogenic activity of its metabolites. PMID- 7497597 TI - Modulation of the in vitro cardiotoxicity of doxorubicin by flavonoids. AB - Cancer therapy with the anthracycline doxorubicin (Dox) is limited by cardiomyopathy, which develops in animals and patients after cumulative dosing. Generation of free radicals by Dox may be involved in this cardiotoxicity. Dox binds strongly to metal ions, especially iron(III). This Dox-metal complex stimulates the generation of free radicals through self-reduction of the complex. We investigated the possibility of inhibiting Dox-induced cardiotoxicity by scavenging of free radicals and/or chelating metal ions. The effects of Dox, both alone and in combination with iron-chelating agents, were studied on inotropy of the isolated mouse left atrium, lipid peroxidation (LPO) in cardiac microsomal membranes, ferricytochrome c (cyt.c3+) reduction, and oxygen consumption. The flavonoids 7-monohydroxyethylrutoside (mono-HER) and 7,3',4' trihydroxyethylrutoside (tri-HER) and the ethylenediaminetetraacetic acid (EDTA) analogue ICRF-198 and its precursor ICRF-187 were used as iron-chelating agents. The latter were used for comparison since ICRF-187 has been reported to inhibit the cardiotoxic effects of Dox both in vitro and in vivo. Only the flavonoids could inhibit the negative inotropic effect of Dox (35 microM) on the mouse left atrium; in the presence of tri-HER (500 microM) the beating force decreased by 18% instead of 50%, whereas mono-HER completely prevented the Dox-induced negative inotropic effect. ICRF-198 and both flavonoids (500 microM) completely inhibited Dox (35 microM)-induced LPO, whereas ICRF-187 provided 65% inhibition. The observation that both cyt.c3+ reduction and oxygen consumption induced by the Dox-iron(III) complex (50/16.6 microM Dox3Fe3+) could be inhibited by superoxide dismutase proved the involvement of superoxide anions (O2-.). The iron-chelating agents (50 microM) inhibited cyt.c3+ reduction by 91% (mono-HER), 43% (tri-HER), and 100% (ICRF-198). Only mono-HER and ICRF-198 (50 microM) were capable of inhibiting the oxygen consumption by 70% and 43%, respectively. It is concluded that flavonoids offer a good perspective for further studies on the prevention of Dox-induced cardiomyopathy. PMID- 7497601 TI - A phase I and pharmacokinetics study of prolonged ambulatory-infusion carboplatin. AB - A total of 18 patients received 6-week ambulatory infusions of carboplatin in groups at dose levels of 14, 28, 35 and 42 mg/m2 per day. The dose-limiting toxicity was myelosuppression. At 42 mg/m2, three of four patients had WHO grade 4 and one of four had grade 3 neutropenia, whereas two patients had grade 3 thrombocytopenia. At 35 mg/m2, two of five patients had grade 3 neutropenia, whereas one had grade 4 and two had grade 3 thrombocytopenia. Non-hematological toxicities were predominantly gastrointestinal, with 3 of 18 patients experiencing grade 3 emesis. Total and ultrafiltrable platinum (UFPt) were assayed by flameless atomic absorption spectrometry in weekly and post-infusion plasma and urine samples. In plasma, levels of total platinum increased throughout the infusion, and the protein binding slowly increased from 60% platinum bound at week 1 to 90% bound by week 4. Although the UFPt level reached a steady state within 1 week, the concentration did not increase with the dose level, remaining at a mean value of 0.58 +/- 0.24 microM. Renal excretion of platinum accounted for 70 +/- 12% of the dose at steady state. There was a high inter-patient variability in both total body clearance of UFPt (range, 83-603 ml/min) and renal clearance (range, 67-390 ml/min). A terminal elimination half life of 13-27 h was noted for post-infusion UFPt. Neutropenia was linearly related to the total daily carboplatin dose, but neither neutropenia nor thrombocytopenia could be related to steady-state UFPt or the UFPt area under the concentration-time curve (AUC). The recommended dose for phase II studies is 28 mg/m2 per day. PMID- 7497600 TI - In vitro effect of r-verapamil on acute myelogenous leukemia blast cells: studies of cytokine secretion and cytokine-dependent blast proliferation. AB - The in vitro effect of the dextroisomer r-verapamil on blast cells derived from patients with acute myelogenous leukemia (AML) was studied. R-verapamil caused a dose-dependent inhibition of AML blast proliferation in the presence of stem-cell factor, leukemia inhibitory factor, interleukin 4, interleukin 6, and interleukin 10 when these cytokines were tested both alone and in different combinations. R verapamil also inhibited the growth of clonogenic AML blast cells. The antiproliferative effect was not specific for AML blast cells, because r verapamil also inhibited cytokine-dependent proliferation of blast cells derived from patients with acute lymphoblastic leukemia. The inhibitory effects of r verapamil and anti-IL1 serum were additive, suggesting that the antiproliferative effect of r-verapamil does not depend solely on inhibition of IL1-mediated effects. Although r-verapamil inhibited spontaneous AML blast proliferation, for a majority of patients it caused only minimal, if any, inhibition of spontaneous cytokine secretion (IL1 alpha, IL1 beta, TNF alpha, IL6) by AML blast cells. Thus, although inhibition of IL1 effects may contribute in certain patients to the antiproliferative effect of r-verapamil, mechanisms other than IL1 inhibition seem to be more important in mediating the effects of r-verapamil. PMID- 7497602 TI - A clinical and pharmacological study of 5-fluorouracil, leucovorin and interferon alfa in advanced colorectal cancer. AB - Modulation of 5-fluorouracil (FUra) using leucovorin (LV) is a standard treatment approach in patients with metastatic colorectal cancer. Modulation of FUra with interferon alfa has also shown some promise. Laboratory data have demonstrated increased cytotoxicity when FUra is combined with both LV and interferon. The current study examined the effects of double modulation of FUra using LV and interferon. Patients with measurable advanced colorectal cancer received bolus FUra 375 mg/m2 plus LV 20 mg/m2 daily for 5 days, repeated every 28 days. Recombinant human interferon alfa-2a, 3 million IU/m2 subcutaneously, was given daily on the days of chemotherapy then three times weekly. There was one complete response and nine partial responses (10/41) seen for an overall response rate of 24% (95% CI 12.0-40.0%). Overall, 70% of patients experienced one or more episodes of nonhematologic toxicity of grade 3 or more. Weight loss was common, with a mean decrease of 2.9 kg over the first two months (P < 0.0001). Improvements in tumor-related symptoms were balanced by increased fatigue and a deterioration in body weight and performance status. There was no evidence of progressive changes in FUra metabolism from interferon usage. PMID- 7497603 TI - Pharmacokinetics of 4'-O-tetrahydropyranyladriamycin given on a weekly schedule in patients with advanced breast cancer. AB - Improved quality of life has gained importance over shortly lasting remissions in yet incurable metastatic breast cancer. Fractionation of drug administration is one of the possible approaches to reduce the concentration-dependent toxicity of anthracyclines. We evaluated the pharmacokinetics of 4'-O tetrahydropyranyladriamycin (THP-ADM) under weekly administration in patients with advanced breast cancer (dose escalation, from 20 to 27 mg/m2 THP-ADM). The concentration-time curves of THP-ADM in plasma were best described by an open three-compartment model [half-life of the first disposition phase (t1/2 alpha), 3.15 min; terminal elimination half-life (t1/2 gamma), 13.9 h] with a mean area under the curve (AUC) of 12.2 ng h ml-1mg-1 m-2, resulting in a mean plasma clearance of 86.9 1h-1 m-2. Metabolism included the formation of Adriamycin (ADM), Adriamycinol (ADM-OH), 13-dihydro-4'-O-tetrahydropyranyladriamycin (THP OH), 7-deoxyadriamycinone (7H-ADn), and 7-deoxy-13-dihydroadriamycinone (7H-ADn OH), with maximal plasma concentrations ranging from 2.8 to 5.5 ng/ml. The mean total amount of cytotoxic anthracyclines excreted into urine, mainly as the parent drug, was 5% of the delivered dose. ADM and ADM-OH, but not the parent drug, were observed in urine at up to 4 weeks after the last therapeutic cycle. There was a significant correlation between the leukocyte nadir under therapy and the AUC of ADM-OH (r = 0.800, P < 0.05). Since no shift in the plasma kinetics was observed from the first to the sixth cycle, the favorable ratio of the AUCs of THP-ADM and ADM after fractionation of THP-ADM suggests lower toxic side effects attributable to ADM. This hypothesis was confirmed in a clinical study, where no severe cardiotoxicity and only mild alopecia were observed in 19 patients. Thus, pharmacokinetics studies might be helpful in both individualization of therapy with THP-ADM and optimization of the administration schedule. PMID- 7497605 TI - Detection of hematogenous dissemination of prostatic cancer by RT-PCR with primers specific for prostate-specific membrane antigen. PMID- 7497606 TI - Overcoming the limitations of artificial clinical specimens: linkage to performance of patients' specimens. PMID- 7497604 TI - Long-term pharmacokinetic behavior of platinum after cisplatin administration. AB - PURPOSE: The platinum concentration in plasma was studied in 19 patients treated by 3 or 4 successive courses of chemotherapy including cisplatin for head and neck cancers. METHODS: Cisplatin was given i.v. daily at 25 mg/m2 by 1-h infusions for 4 days every 3 weeks. Total and ultrafiltrable platinum were measured in plasma using an inductively coupled plasma mass spectrometry (ICPMS) technique. RESULTS: A progressive accumulation of total platinum in plasma was observed with consecutive infusions. The mean (+/- SD) total plasma platinum level detected at the end of cisplatin infusion was 1134 +/- 234, 1407 +/- 268, and 1618 +/- 282 micrograms/l at the end of the first, second, and third courses, respectively. The minimal platinum concentration measured before the second and third courses also increased to 221 +/- 59 and 309 +/- 76 micrograms/l, respectively. The steady state was not reached before the third course. However, differences in the evolution of platinum plasma levels were found among the 19 patients. In 14 patients the pharmacokinetics of platinum was characterized by low initial levels, a progressive accumulation, and a long terminal half-life with a very late steady state. In 5 patients, the pharmacokinetic behavior of platinum was different: platinum levels were directly high, without progressive accumulation, the steady state being reached as early as the first course. Significant levels of ultrafiltrable platinum were found throughout the treatment, even during the intervals between courses with this very sensitive analytical method. A close equilibrium between ultrafiltrable and total platinum (ratio, 6%) persisted for as long as 3 weeks after cisplatin administration. DISCUSSION: These results underline the importance of individual differences in platinum metabolism. The relationship between total and ultrafiltrable platinum are discussed. PMID- 7497607 TI - Application of a low-light imaging device and chemiluminescent substrates for quantitative detection of viral DNA in hybridization reactions. AB - In this quantitative dot-blot hybridization assay for detecting B19 parvovirus DNA, we used three different chemiluminescent substrates [adamantyl-1,2-dioxetane phenyl phosphates (PPD and the new PPD-Plus) and the chloro-5-substituted adamantyl-1,2-dioxetane phosphate (CSPD) plus Emerald enhancer] and a high performance, low-intensity-light imaging luminograph apparatus. The hybridization test uses digoxigenin-labeled DNA probes, which are immunoenzymatically revealed by anti-digoxigenin Fab fragments conjugated with alkaline phosphatase. All the detection systems with the various chemiluminescent substrates gave sensitive and reproducible results for calibrators and positive or negative reference clinical samples, with high reproducibility (CV 4-17%). The signal was measured after 45 min of incubation. The luminograph apparatus could detect 10 fg of homologous DNA with the PPD-Plus substrate, whereas the detection limit with the CSPD and PPD substrates was 20 fg and 20-50 fg, respectively. Analysis of 26 samples with the three substrates showed good sensitivity and specificity for viral detection. PMID- 7497609 TI - Detection of prostate-specific antigen mRNA by reverse transcription polymerase chain reaction and time-resolved fluorometry. AB - We have developed a time-resolved fluorometric hybridization assay for detecting prostate-specific antigen (PSA) mRNA amplified by reverse transcription polymerase chain reaction. During PCR, digoxigenin-11-dUTP is incorporated into the amplified product. An oligonucleotide internal to the primers is used as a specific probe, being biotinylated and captured on streptavidin-coated microtiter wells. Denatured PCR product hybridizes with the probe, and the hybrids are detected with an alkaline phosphatase-labeled antidigoxigenin antibody. We used the phosphate ester of fluorosalicylic acid as the substrate. The fluorosalicylate produced forms a highly fluorescent ternary complex with Tb(3+) EDTA, which we can measure by time-resolved fluorometry. A signal-to-background ratio of 10 was obtained when 160 PSA cDNA molecules were present in the preamplification sample. Also, mRNA corresponding to one LNCaP cell in the presence of 10(6) PSA-negative cells can be detected (signal-to-background ratio of 3.1). Samples containing 100, 1000, and 50,000 LNCaP cells gave CVs of 12.4%, 4.9%, and 6.8%, respectively (n = 10). PMID- 7497608 TI - Enhanced detection of hematogenous circulating prostatic cells in patients with prostate adenocarcinoma by using nested reverse transcription polymerase chain reaction assay based on prostate-specific membrane antigen. AB - We report the development of a new sensitive nested reverse transcription polymerase chain reaction (RT-PCR) assay, using primers derived from the prostate specific membrane antigen (PSM) cDNA sequence, to detect an hematogenous spread of prostate adenocarcinoma cells. In 60 patients with a biopsy-proven prostate cancer, PSM and PSA RT-PCR detected circulating prostate cells in 40 and 20 patients, respectively. In pT4 M+ and pT3 M+ disease patients, nested PSM primers detected cells in 28 of 33 patients (85%), whereas nested PSA primers detected cells in 17 of 33 (51%). In patients with organ-confined cancer spread (pT2a and pT2b patients) before radical prostatectomy, nested PSM RT-PCR detected circulating prostatic epithelial cells in 6 of 17 patients (35%), which suggests that an hematogenous spread of prostate cells may occur early in prostate cancer history. Altogether, these results suggest that the detection of PSM-expressing cells in blood may predict the development of cancer in patients without clinically apparent prostate cancer. Nevertheless, the potential application and the clinical significance of detection of hematogenous prostate cells through the use of nested PSM primers need an extensive longitudinal study. PMID- 7497610 TI - Cardiac troponin-I is not expressed in fetal and healthy or diseased adult human skeletal muscle tissue. AB - Cardiac troponin-I (cTnI) is not found in sera of patients with skeletal muscle disease in the absence of myocardial injury. It is not known, however, whether trace amounts of cTnI are expressed in regenerating human skeletal muscle, as has been observed with creatine kinase MB. Using immunohistochemical and biochemical techniques, we investigated cTnI expression in various human muscle tissues: human heart tissue (n = 5), normal adult skeletal muscle (n = 3), and fetal heart (n = 3) and skeletal muscle (n = 3) obtained, respectively, during heart transplant, from autopsy, or from a tissue bank. Specimens from diagnostic tissue biopsies were used as diseased skeletal muscle: polymyositis (PM), n = 13; Duchenne muscular dystrophy (DMD), n = 6. Frozen sections 8 microns thick were stained immunohistochemically for either cTnI or TnI (cardiac or skeletal) by using monoclonal antibodies (MAb) 2B1.9 (cTnI specific) or 3C5.10 (reactive with all TnI isoforms), respectively. cTnI was measured in tissue homogenates by an immunofluorometric assay. Cardiac muscle was stained by both MAbs. Normal fetal and adult skeletal muscle, and samples from all of the PM and DMD patients, stained only with the nonspecific MAb (3C5.10), confirming the sole presence of skeletal TnI. No cTnI was detectable by immunoassay in any skeletal muscle sample. We conclude that cTnI is not expressed in human skeletal muscle during development or during regenerative muscle disease processes such as PM or DMD. PMID- 7497611 TI - Plasma alpha-glutathione S-transferase assessed as a marker of liver damage in patients with chronic hepatitis C. AB - alpha-Glutathione S-transferase (alpha-GST; EC 2.5.1.18) has been advocated as a better marker of hepatocellular damage than the transaminases in toxic and autoimmune hepatitis. We have assessed the potential interest of plasma alpha-GST determination in 94 anti-hepatitis C virus-positive patients with histologically proven chronic hepatitis C (34 women, 60 men, ages 40.0 +/- 11.9 years). Blood samples were assayed for aspartate aminotransferase (AST), alanine aminotransferase (ALT), gamma-glutamyltransferase, alkaline phosphatase, and alpha-GST on the same day a liver biopsy was performed. alpha-GST concentrations were significantly above reference values in 64% of patients (compared with 58% for AST, 68% for ALT), and this increase was seen in 52% of patients with normal values for transaminases and a Knodell score > 3. Furthermore, there was a significant correlation between alpha-GST and lobular necrosis score (r = 0.31; P < 0.01). Our findings suggest that association of plasma alpha-GST with ALT may improve the biochemical assessment of liver damage in patients with chronic hepatitis C. PMID- 7497612 TI - Time-resolved immunofluorometric assay of p53 protein. AB - A common feature of human tumor tissue is mutant p53 protein accumulation. Here we evaluate a new "sandwich" immunoassay for p53 protein incorporating modifications to a previously reported method, including the use of microtiter plates coated directly with the anti-p53 monoclonal antibody DO-1, a detergent- and mouse serum-containing sample diluent, and a labeled secondary antibody diluent containing goat serum. The use of CM-1 antiserum to probe the immunocaptured p53 and the detection of bound complexes by a labeled secondary antibody allows coupling to a time-resolved fluorescence detection system. The new assay yielded p53 concentrations comparable with those by the previous assay for breast tumor cytosols (n = 198), nondiseased breast tissues (n = 70), and five transformed cell lines, but showed differences in p53 values measured in sera from patients without cancer (n = 78). These serum differences were found to reflect nonspecific interferences affecting the original method, which implies that the new immunoassay has improved specificity for serum p53 quantification. PMID- 7497613 TI - Variation in measurement of prostate-specific antigen: importance of method and lot variability. AB - Equivalence between Hybritech Tandem and Abbott IMx PSA methods have been reported by some but not all previous investigators. To determine reasons for these differing conclusions, we measured serum PSA with three different lots each of IMx and Tandem-E kits. Overall, mean IMx results were significantly lower than Tandem-E results; however, for selected sera, the IMx results were consistently higher than the Tandem-E results. Lot-to-lot differences for the IMx method were significantly greater than those with the Tandem-E method. Most IMx/Tandem-E lot to-lot comparisons had linear regression slopes that differed significantly from 1.0, but some did not. Conclusions concerning the equivalence of the IMx and the Tandem-E methods can be influenced both by variations in the proportions of free PSA in sera in tested populations and by lot-to-lot differences in the IMx method. PMID- 7497614 TI - Automated assay of oxygen radical absorbance capacity with the COBAS FARA II. AB - Reactive oxygen species are of interest in biology and medicine because of evidence relating them to aging and disease processes. A relatively simple but sensitive and reliable method for quantifying the oxygen radical absorbance capacity (ORAC) of antioxidants in biological tissues has been automated for use with the COBAS FARA II centrifugal analyzer with a fluorescence-measuring attachment. In this assay, beta-phycoerythrin (beta-PE) is used as an indicator protein, 2,2'-azobis(2-amidinopropane) dihydrochloride (AAPH) as a peroxyl radical generator, and 6-hydroxy-2,5,7,8-tetramethylchroman-2-carboxylic acid (Trolox) as a calibrator for antioxidant activity. This assay is unique because the reaction goes to completion so that both inhibition time and inhibition degree are considered in quantifying ORAC (micromoles of Trolox equivalent per liter or per gram of tissue). This method can be used not only for serum but also other tissue and food samples and is suitable for application to a range of nutritional and clinical conditions. PMID- 7497616 TI - Cobas Integra: clinical laboratory instrument with continuous and random-access capabilities. AB - The Cobas Integra from Roche Diagnostic Systems is a new clinical laboratory analyzer with continuous and random-access features for routine chemistries, specific proteins, electrolytes, hormones, therapeutic drugs, and drugs of abuse. The system maintains 68 test-specific reagent cassettes on board, along with multiple ion-selective electrodes (ISEs) for electrolyte determinations. This gives the Cobas Integra the capability of analyzing as many as 72 analytes without having to load additional reagents. We describe the basic analyzer configuration and the subsystems for absorbance, fluorescence polarization, and ISE measurements. Performance characteristics for precision, methods comparison, and on-board stability are given for assays representative of the various test groups. The 29 Cobas Integra tests evaluated in the present study show good agreement (r > or = 0.98 and slopes generally 0.90 to 1.12) with the respective methods available on either the Olympus AU5000, Hitachi 911 or 717, Behring BNA, or Abbott TDx systems. Total assay precision (CV) ranged from 0.8% to 8.5%, and calibration curves were stable for as long as 20 weeks. Test throughput, which is dependent on pipetting sequence, was determined to be up to 600 tests per hour without ISE and up to 750 tests per hour with ISE; the time to first result was 2.0-10.0 min. PMID- 7497615 TI - Enzymatic method for determining ketone body ratio in arterial blood. AB - We have developed a new, sensitive, and rapid method for measuring the ketone body concentration in arterial blood and determining the arterial blood ketone body ratio. The procedure involves the sequential use of the enzymes 3 hydroxybutyrate dehydrogenase (3-HBDH; EC 1.1.1.30) and NADH oxidase, followed by a color-generating reaction with the hydrogen peroxide produced by the oxidase reaction. The amount of oxidized chromogen produced is proportional to the 3 hydroxybutyrate (3-HBA) concentration. The acetoacetate (AcAc) concentration is obtained after complete conversion of the AcAc to 3-HBA, in the presence of 3 HBDH. The total 3-HBA concentration is measured and then subtracted from the total ketone body concentration to give the AcAc concentration. This procedure may be applied to plasma samples and the absorbance change measured with an automated chemistry analyzer. Ketone body concentration may be determined over the range 0 to 400 mumol/L. The analysis takes approximately 12 min and requires only 30 microL of plasma. PMID- 7497618 TI - Intracellular magnesium content of mononuclear blood cells and granulocytes isolated from leukemic, infected, and granulocyte colony-stimulating factor treated patients. AB - The intracellular magnesium (Mg) concentration of granulocytes and mononuclear blood cells (MBCs) was determined in cells isolated from patients with several disorders. The mean (+/-SD) Mg content of MBCs isolated from patients diagnosed with lymphocytic leukemia, myelocytic leukemia, or infection; from patients treated with granulocyte colony-stimulating factor (G-CSF); and from healthy volunteers (control group) was 2.3 (+/-0.6), 3.3 (+/-0.5), 4.1 (+/-0.8), 3.9 (+/ 0.4), and 3.9 (+/-0.6) fmol/cell, respectively. The Mg content of MBCs isolated from patients with lymphocytic and myelocytic leukemia, but not those from patients with infection or receiving G-CSF treatment, were significantly lower (P < 0.001) than those from the control subjects. The mean Mg concentration of granulocytes obtained from lymphocytic leukemia, myelocytic leukemia, infection, and G-CSF patients and from the control group was 3.2 (+/-0.9), 3.4 (+/-0.5), 3.8 (+/-0.6), 4.5 (+/-0.6), and 4.6 (+/-0.6) fmol/cell, respectively. Granulocytes isolated from leukemic and infectious patients yielded lower intracellular Mg concentrations (P < 0.005) than those from patients receiving G-CSF and the control group. This study demonstrates that intracellular Mg content is altered in several pathological states. Several factors, including depleted Mg stores or altered intracellular Mg binding sites, could be responsible for these changes. Apparently, intracellular Mg content may be of use in assessing total body Mg status. PMID- 7497617 TI - Quantitative determination of high-, low-, and very-low-density lipoproteins and lipoprotein(a) by agarose gel electrophoresis and enzymatic cholesterol staining. AB - Quantification of lipoprotein cholesterol was performed by enzymatic staining of cholesterol in a new agarose gel electrophoresis method that allows the separation of LDL, VLDL, HDL, and lipoprotein(a) [Lp(a)]. Lp(a) shows an electrophoretic mobility clearly distinct from VLDL and HDL. The total CVs of lipoprotein cholesterol varied between 2.7% and 3.9% for LDL, 7.8% and 23.2% for VLDL, 5.2% and 9.5% for HDL, and 6.8% and 16.4% for Lp(a). Comparison of LDL-, VLDL-, and HDL-cholesterol concentrations with the results of a combined ultracentrifugation and precipitation technique gave correlation coefficients of 0.961, 0.947, and 0.918, respectively; comparison of Lp(a)-cholesterol values with those of a nephelometric Lp(a) assay gave r = 0.906. The new electrophoretic assay has several advantages: It allows the quantification of Lp(a)-cholesterol; VLDL-cholesterol is not affected by Lp(a)-cholesterol; and the LDL-cholesterol fraction does not contain Lp(a)-cholesterol, as happens with LDL-cholesterol determined by ultracentrifugation and precipitation. PMID- 7497619 TI - Artificial icteric plasmas: unreliable indicators for interference with creatinine assay on Beckman CX3. PMID- 7497620 TI - Cholesterol index of hair in normal and hypercholesterolemic mice. PMID- 7497623 TI - Improved method for determining fetal hemoglobin (HbF) by alkali denaturation. PMID- 7497622 TI - Humalog, a rapidly absorbed analog of human insulin, determined with Coat-A-Count insulin radioimmunoassay. PMID- 7497621 TI - Amplification refractory mutation system to identify mutations in cystathionine beta-synthase deficiency. PMID- 7497624 TI - Ion capture assay for folate with the Abbott IMx analyzer. PMID- 7497625 TI - Three routine methods for serum cortisol evaluated by comparison with an isotope dilution gas chromatography-mass spectrometry method. PMID- 7497626 TI - Direct measurement of HDL cholesterol preferable to precipitation method. PMID- 7497629 TI - Test characteristics of cerebrospinal fluid tumor markers. PMID- 7497628 TI - Test characteristics of cerebrospinal fluid tumor markers. PMID- 7497627 TI - Effects of preanalytical temperature on thyrotropin measurements by Ciba Corning ACS:180. PMID- 7497630 TI - Identification of mutated p53 in cancer by non-gel-sieving capillary electrophoretic SSCP analysis. PMID- 7497631 TI - Antioxidant activity of resveratrol in red wine. PMID- 7497632 TI - Severe hypouricemia: biochemical expression of asymptomatic xanthinuria. PMID- 7497633 TI - Anti-solid phase antibodies interfering in labeled-antibody assays of free thyroid hormones. PMID- 7497634 TI - Biomarkers in Environmental Toxicology. The Arnold O. Beckman/IFCC European Conference on Environmental Toxicology. Cannes, France, 1-3 June, 1995. Proceedings and abstracts. PMID- 7497635 TI - Biomarkers in epidemiology. AB - A biomarker is a measurable event occurring in a biological system, such as the human body. In environmental epidemiology, a biomarker represents a subclinical and reversible change; it is not a diagnostic test, but an indicator that an early change has occurred that could later lead to clinical disease. Although some biomarkers may belong to more than one class, they are often separated into biomarkers of exposure, biomarkers of effect, and biomarkers of susceptibility. Biomarkers can be used to classify and quantify environmental exposures and their related effects, and many methods may be applicable in toxicological experiments as well as in epidemiology. Accordingly, biomarker epidemiology is undergoing rapid development and expansion and is becoming one of the most promising areas of environmental research. Although expanded applications should be encouraged, many biomarkers are poorly characterized, and attention should be paid to defining their properties in detail. PMID- 7497636 TI - Biomarkers as tools in human health risk assessment. AB - Evaluation of occupational or environmental risk due to exposure to chemicals requires sufficient information on the toxic profiles, mechanisms of action, toxicokinetics, dose-response relation, exposure, and the target dose. Usually exposure is estimated by measuring concentrations of the agent in air, food, water, soil, dust, or other media with which a population or an individual is in contact. However, this external exposure is only a rough estimate for the internal exposure (agent dose or its metabolite at the critical target in the organism). Factors of influence are bioavailability of the chemicals, variations in concentrations and routes of exposure, physical activity, and individual variation in rates of metabolism, distribution, and excretion. All these affect the concentration of the toxic agent at the critical target, which is the most precise information for risk assessment. Thus, internal exposure is best measured by determining the concentration of the toxicant or its ultimate metabolite at the critical site in the target organ or by determining adducts with cellular macromolecules such as proteins, amino acids, DNA, or its bases. The latter are easily available in experimental toxicology from animal experiments but only occasionally from humans. For health surveillance such data usually are not available, because they require invasive procedures such as biopsies. Therefore, more accessible body fluids or tissue are used, such as blood, urine, or adipose tissue, or adducts with macromolecules such as albumin or hemoglobin in the blood, DNA adducts in peripheral lymphocytes, or altered DNA bases in urine such as 8-hydroxyguanine. All of these are indicators for exposure, whereas risk can only be estimated if the correlation between their deviations from normal and the dose-response at the critical target is known. PMID- 7497637 TI - Significance of individual sensitivity to chemicals: elucidation of host susceptibility by use of biomarkers in environmental health research. AB - Biomarker research has become the predominant theme for study of human dose-host response relations to environmental chemicals. Increasing interest has been focused on identifying markers for host susceptibility, with mixed results. Efforts to identify markers for host variability in carcinogenic risk, on the basis of theoretical knowledge of carcinogen metabolism, have been disappointing. New work in the area of acquired risk modifiers, such as nutritional status, is theoretically attractive, but results have been limited. Impressive achievements have been made in the area of immunological variability, which may elucidate the molecular basis of as well as provide practical biomarkers for several diseases. The problem of multiple chemical sensitivities, on the other hand, has proved refractory to biomarker research, reflecting inadequate knowledge of the mechanism and inappropriate application of biomarker methods. PMID- 7497638 TI - Cholinesterase inhibition: complexities in interpretation. AB - Cholinesterases are measured to assess exposures to or effects of organophosphorus esters and carbamates. Plasma butyrylcholinesterase is usually most sensitive to inhibitors, but it has no known physiological function(s); its inhibition reflects exposure. The physiological function of erythrocyte acetylcholinesterase (AChE) also is not known, but the enzyme is the same as that involved in synaptic transmission and its measurement is used to mirror effects on the nervous system. Erythrocyte AChE has large inter- and intraindividual variation, and small changes are detectable by comparison with preexposure values. The relation between inhibition of erythrocytes and nervous tissue AChE depends on the pharmacokinetics of inhibitors. Usually, erythrocyte AChE inhibition overestimates that in the nervous system. Pharmacodynamic factors such as spontaneous reactivation and aging of inhibited enzyme should also be considered in assessing AChE inhibition. Other factors, such as timing of measurement, add complexity because erythrocyte AChE inhibition persists longer than that in the nervous tissues. Cholinergic transmission might also be impaired because of direct effects of organophosphorus esters and carbamates on receptors. PMID- 7497639 TI - Lipid peroxidation and antioxidants as biomarkers of tissue damage. AB - Disturbance of the balance between the production of reactive oxygen species such as superoxide; hydrogen peroxide; hypochlorous acid; hydroxyl, alkoxyl, and peroxyl radicals; and antioxidant defenses against them produces oxidative stress, which amplifies tissue damage by releasing prooxidative forms of reactive iron that are able to drive Fenton chemistry and lipid peroxidation and by eroding away protective sacrificial antioxidants. The body has a hierarchy of defense strategies to deal with oxidative stress within different cellular compartments, and superimposed on these are gene-regulated defenses involving the heat-shock and oxidant stress proteins. PMID- 7497640 TI - Induced gene transcription: implications for biomarkers. AB - Numerous xenobiotics regulate cellular functions by altering transcription of target genes. Use of sensitive and specific biomarkers based on gene transcript levels may help clarify the shape of the dose-response curve in the low-dose region associated with human exposures to environmental concentrations of chemicals. We have quantified gene transcription induced by 2,3,7,8 tetrachlorodibenzo-p-dioxin in both animal models and humans with the use of Northern analysis and PCR-based methods. In addition, we describe a rapid and sensitive in vitro assay that we have used to screen chemicals and biological samples for their ability to alter gene transcription. Whereas some of the endpoints in our studies such as cytochrome P-450 1A1 are predictive indicators of exposure and dose, other gene responses such as growth factors are more complex and represent a critical event, progression, or adaptation to a pathological alteration. In conclusion, measurement of toxicant-induced gene transcription will contribute to the usefulness of biomarkers in addressing issues of human health and environmentally induced disease. PMID- 7497641 TI - Macromolecular adducts caused by environmental chemicals. AB - We describe three biomonitoring studies in which hemoglobin (Hb) adducts were used as biochemical markers to assess indirectly the target dose of genotoxic chemicals. We monitored the exposure to 1,3-butadiene in occupationally exposed workers and in two control groups by analyzing the adducts formed by the reaction of the first activation product, butadiene monoepoxide, with the terminal valine of Hb; we also measured hydrolyzable adducts formed by the reaction of metabolically formed nitroso derivatives with Hb from five selected nitropolycyclic aromatic hydrocarbons (1-nitropyrene; 2-nitrofluorene, 3 nitrofluoranthrene, 6-nitrochrysene, and 9-nitrophenanthrene) in coke oven workers of different job categories and control workers of the same geographical area. We detected hydrolyzable adducts from monocyclic nitroarenes in blood from individuals living in a contaminated area where explosives had been produced and from controls. The contaminants considered were 2,4,6-trinitrotoluene; 2,4- and 2,6-dinitrotoluene; and 1,3-dinitrobenzene. Differences between groups were significant, but interindividual variation was great and back-ground exposures must be considered. PMID- 7497642 TI - Detection of point mutations in T lymphocytes. AB - We exposed experimental animals to a series of alkylating agents that induced mutations at the X-linked hprt gene of T lymphocytes. We then isolated the mutant cells and analyzed the molecular nature of the mutations by amplification of hprt cDNA sequences with the use of reverse transcriptase PCR followed by DNA sequence analysis, and then correlated the mutational spectra obtained to the spectra of DNA adducts caused by the alkylating agents used. The nature of the base-pair changes causing the mutations was characteristic for the reaction pattern of the genotoxic agent with DNA. However, we also found a clear influence of DNA repair processes; i.e., in those cells that were able to remove certain types of DNA damage, the class of mutations expected from that type of damage was reduced. PMID- 7497643 TI - Serum oncoproteins in asbestosis patients. AB - Using ELISAs, we determined the concentrations of transforming growth factor alpha (TGF-alpha), the extracellular domain of the erbB-2 receptor (erbB-2 ECD), and mutant p53 protein in stored serum samples of asbestosis patients with and without cancer and control subjects (without asbestosis or cancer). The percentage of individuals in these three groups with increased serum concentrations of TGF-alpha, erbB-2 ECD, and mutant p53, respectively, were: asbestosis patients with cancer, 36%, 16%, 19%; asbestosis patients without cancer, 38%, 19%, 6%; control subjects, 0%, 5%, 10%. Although differences in serum positivity for these oncoproteins were apparent among these groups, the differences did not achieve statistical significance (P > 0.05). In several of the cancer cases, increased concentrations of TGF-alpha, erbB-2 ECD, and mutant p53 were also detected in the stored serum samples collected years before the clinical diagnosis of disease. PMID- 7497644 TI - Variability in DNA repair and individual susceptibility to genotoxins. AB - DNA repair is an important mechanism of cellular protection from the effects of genotoxic chemicals. Although extensive evidence from studies in experimental systems indicates that variation in DNA repair can significantly influence susceptibility to genotoxins, corresponding studies in human populations are so far limited, mainly because of methodological difficulties. One system, using observations of the accumulation and repair of DNA damage in cancer patients treated with alkylating cytostatic drugs, has provided useful information for assessing the effects of interindividual variation in DNA repair activity on the induction of genotoxic effects in humans. The most detailed studies of this kind have been carried out on patients with cancer (i.e., Hodgkin disease, malignant melanoma) treated with the methylating cytostatic drugs procarbazine or dacarbazine; these studies have provided detailed information on dose-response relationships. They have also demonstrated the protective role of the repair enzyme O6-alkylguanine-DNA alkyltransferase against the accumulation of the premutagenic methylated DNA lesion O6-methylguanine in patients' DNA. Given the strong evidence that exposure of the general population to environmental methylating agents may be extensive, as indicated by the frequent discovery of methylated DNA adducts in human DNA, data on DNA damage and repair in alkylating drug-treated patients and their modulation by host factors may prove useful in efforts to assess the possible carcinogenic risks posed by exposure to environmental methylating agents. PMID- 7497645 TI - DNA repair and epidemiology of basal cell carcinoma. AB - In a molecular epidemiological study of DNA repair, host reactivation assay was used to measure the DNA repair capacity of cryopreserved lymphocytes from 88 primary basal cell carcinoma (BCC) patients and 135 cancer-free controls. In this study population, reduced repair of ultraviolet radiation-induced DNA damage contributed to the risk of sunlight-induced BCC. A family history of BCC is associated with low DNA repair. Repair of ultraviolet radiation-damaged DNA declines at a rate of approximately 1%/year in noncancerous controls. Reduced DNA repair is more likely seen in young BCC patients, indicating that BCC is a premature aging disease of the skin. The persistence of photochemical damage because of reduced repair results in point mutations in the p53 gene and allelic loss of the nevoid BCC gene located on chromosome 9q. Xeroderma pigmentosum appears to be a valid paradigm for the role of DNA repair in BCC in the general population. PMID- 7497646 TI - Polymorphisms in drug-metabolizing enzymes as modifiers of cancer risk. AB - The identification of low-penetrance genes, the polymorphisms of which increase an individual's risk of developing cancer, are likely to be extremely important in the general population. In this report we analyzed two genes involved in detoxification. In a number of loci, we identified polymorphic variation correlating with the expression of the gene product. We analyzed two such loci, the cytochrome P-450 gene CYP2D6 and the N-acetyltransferase 2 (NAT2) genes, in patients with bladder and colon cancer, respectively. We observed no statistically significant associations between the control and cancer populations; however, there was a small increase in heterozygote number in bladder cancer. PMID- 7497647 TI - Assessment of immunotoxic effects in humans. AB - The immunotoxic effects of chemicals are varied and markedly different depending on the underlying pathogenesis, namely, direct immunotoxicity (including immunosuppression, immunodepression, and immunostimulation), hypersensitivity, and autoimmunity. A large number of immunological endpoints and functional assays have been proposed for use as biomarkers of immunotoxicity, but they often lack sensitivity or are poorly standardized, so that their relevance in assessing immunotoxic effects in humans is at best ill established. Examining sentinel immunopathological events in individuals with a defined history of chemical exposure is another approach, presumably more cost-effective at the present time. A multicenter collaboration is mandated, however, because these events are rare. We expect that progress in new technologies, e.g., molecular biology, will provide the sensitive and reliable biomarkers of immunotoxicity that are currently lacking. PMID- 7497648 TI - Markers of neurotoxicity: from behavior to autoantibodies against brain proteins. AB - Evidence of potentially neurotoxic exposures may be obtained in peripheral indicator media, but molecular or cellular evidence of neurotoxic effects has not been as readily available, primarily because the nervous system of living humans is beyond the reach of direct measures. Although there is limited evidence that molecular changes in the blood can provide information about neurotoxicity, several new approaches are being investigated. One is that the immune system may preserve evidence of damage to the nervous system. Debris from damaged cells in the nervous system may present as antigens, giving rise to autoantibodies, which may be detectable in blood for a long time after injury. Progress will depend on more control experiments and clarification of confounding variables. Validation of new molecular markers must go hand in hand with documentation of impaired function, most commonly measured as behavioral or neurophysiological changes. These noninvasive markers will be measured with greater sensitivity and precision, thanks to innovative computer technology. The practical advantages of the new markers may be as important as their contribution to our understanding of the mechanisms by which the nervous system defends against chemical insult. PMID- 7497651 TI - Do environmental estrogens contribute to the decline in male reproductive health? AB - Several observations suggest that male reproductive health has been declining since World War II in many countries. The incidence of testicular cancer, hypospadias, and cryptorchidism has been increasing and semen quality has been decreasing, and these may have a common etiology. Treatment of several million pregnant women with the synthetic estrogen diethylstilbestrol led to an increase in these conditions among the sons of these women. These abnormalities probably arise during fetal development. The similarity between these effects and the adverse change in male reproductive development and function raised the question of whether the adverse changes are attributable to altered exposures to estrogenic and other endocrine-disrupting agents during fetal development. We speculate that alteration in exposure to estrogen in the past half-century may have caused the changes in male reproductive health. PMID- 7497649 TI - Manifestations of chemically induced liver damage. AB - Possible liver damage induced by chemicals or drugs must be detected early during drug development or industrial exposure, although damage is still difficult to predict, especially when immunotoxicity is involved. Liver toxicity may result from cytolysis, steatosis, cholestasis, phospholipidosis, or vascular lesions, most the outcome of a disadvantageous balance between chemicals or metabolites vs protective mechanisms, resulting from chemical dosage, genetic factors, or the immunoallergic status of the patient. Drug metabolism, lipid peroxidation, and thiol oxidation are frequently involved in liver toxicities. Classical guidelines in toxicology propose many methods for liver toxicity assessment: histology; chemical changes in hepatic tissue (lipids, glutathione, enzymes); physiological changes in biosynthesis (proteins, glycoproteins); excretion function (fructose); drug metabolism; and concentrations of related enzymes (alanine aminotransferase, aspartate aminotransferase, alkaline phosphatase, and gamma-glutamyltransferase) in blood. In vitro studies in human or animal hepatocytes or tumor-derived cell lines are useful in detecting hepatocellular lesions by cell viability, glutathione concentration, amount of lactate dehydrogenase released, cellular ATP, morphology (blebs), and drug metabolism. PMID- 7497650 TI - Development of a marker of estrogenic exposure in human serum. AB - The deleterious, disruptive effects of estrogen mimics on the endocrine system were discovered after the compounds were released into the environment. Their chemical structure does not obviously resemble that of steroid hormones; hence, their estrogenic effects were totally unexpected. In addition to occupational exposures, environmental estrogens may have played a role in decreasing the quantity and quality of human semen during the last 50 years and in increasing the incidences of testicular cancer and cryptorchidism in men and breast cancer in women and men in industrialized countries. Testing the environmental estrogen hypothesis will require developing appropriate biomarkers of exposure and measuring these biomarkers at developmental points where exposure is critical. We report the ongoing development of a method to extract and separate xenoestrogens from ovarian estrogens with human serum as a source, followed by determination of xenoestrogen concentration by a bioassay. We also critically assess bioassays currently available to measure the cumulative effect of xenoestrogens, e.g., (a) the E-SCREEN assay, which measures the proliferative effect of estrogens on their target cells, and (b) the induction by estrogens of specific gene products, such as progesterone receptor and pS2. PMID- 7497652 TI - Biomarkers in environmental toxicology: state of the art. PMID- 7497653 TI - Revision total knee arthroplasty after failed unicompartmental knee arthroplasty or high tibial osteotomy. AB - A retrospective matched-pair comparative analysis was done between 30 total knee arthroplasties following failed high tibial osteotomies and 30 total knee arthroplasties following failed unicompartmental knee arthroplasties. The groups were matched according to age, gender, type of prosthesis, primary disease, and length of followup. A minimum followup of 2 years was required for inclusion in the study, and the average followup was 3.8 years (range, 2-9 years). The Knee Society Knee Score for the high tibial osteotomy group was significantly higher than that for the unicompartmental arthroplasty group. More osseous reconstructions were required in the unicompartmental revisions. Difficulty with exposure was not significantly greater in the osteotomy group. Rates of component loosening were not significantly different between the groups. A failed unicompartmental knee arthroplasty and a failed high tibial osteotomy can be revised successfully to a total knee arthroplasty. The results confirm that revisions after unicondylar arthroplasty and high tibial osteotomy are technically demanding. In this series, the results of total knee arthroplasty following unicompartmental knee arthroplasty approached but did not equal those obtained after high tibial osteotomy. PMID- 7497654 TI - Accuracy of intramedullary versus extramedullary tibial alignment cutting systems in total knee arthroplasty. AB - The postoperative standing 51-inch radiographs of 352 total knee arthroplasties were reviewed. The angle formed by the tibial tray to the mechanical axis of the tibia in the coronal plane was measured. The mean deviation from a perpendicular cut to the tibial mechanical axis was not significantly different between tibial cuts made with extramedullary alignment and with intramedullary alignment. With extramedullary alignment, 92% of the cuts were +/- 4 degrees of the ideal 90 degrees cut (perpendicular to the mechanical axis), whereas 94% of the cuts with intramedullary alignment were within 4 degrees of ideal. An analysis of the cuts that erred > or = 4 degrees with intramedullary alignment showed that for the majority the intra-medullary guide rod, if passed down the middle of the tibial shaft in the coronal plane, directly led to the less than ideal cut. In these tibias, bowing deformity in the coronal plane made passage of the intramedullary rod parallel to the tibial mechanical axis impossible. PMID- 7497655 TI - The anatomy of the tibial intramedullary canal. AB - An autopsy study was done to determine the relationship of the tibial plateau and the intramedullary canal of the tibia. Rods were placed in 10 adult anatomic specimen tibias. Computed axial tomography scans were obtained of the proximal tibia at 1 cm, 1.5 cm, 2 cm, and 2.5 cm from the most proximal plateau. Based on these sections, the position of the tibial canal was measured relative to the plateau. The center of the tibial canal on the plateau varied from 28% to 53% of the depth of the tibia from the anterior cortex and from 37% to 56% of the width of the tibia from the medial cortex. Actual measurements ranged from 15 mm anterior to the center to 1.5 mm posterior to the center and 8 mm medial to the center to 4.5 mm lateral. Because of this variability in location of the tibial canal, a mobile stem attachment site on the tibial tray or an offset stem may be beneficial for some patients having total knee arthroplasty. PMID- 7497656 TI - The efficacy of intramedullary femoral alignment in total knee replacement. AB - The angle formed between the femoral anatomic axis and the femoral prosthesis was measured on radiographs of 201 total knee arthroplasties. The radiographs were taken on a 51 inch x 14 inch cassette and had to have satisfactory centering and minimal rotation to be included. All total knee arthroplasties had intramedullary femoral alignment. The mean femoral component angle (relative to the femoral anatomic axis) was 6.1 degrees valgus, with a standard deviation of 2 degrees and a range from 2 degrees to 12 degrees valgus. Seventeen knees (8.5%) had a femoral component angle that was < or = 2 degrees valgus or > or = 10 degrees valgus. In 4 knees, medial femoral bowing of the distal third of the femoral shaft caused an excessive valgus cut. In 7 knees, a capacious femoral canal was present that potentially allowed for divergence of the relatively undersized guide rod in the canal. Overall, 8.5% of cuts were believed to be less than ideal. Preoperative radiographs of the entire femur should identify patients in whom errors are more likely and allow for templating the amount of distal femoral bone to be resected. Extra caution is needed in bowed femora and in capacious femoral canals. PMID- 7497657 TI - Muscle flaps in total knee arthroplasty. AB - The results of 12 patients who had been treated with muscle flap coverage for compromised soft tissues associated with total knee arthroplasty were studied. Five latissimus dorsi free flaps, 6 medial gastrocnemius rotational flaps, and 2 rectus abdominis free flaps were done in 12 knees. The authors used and classified 3 types of muscle flaps: (1) prophylactic soft tissue coverage, done before definitive reconstruction; (2) treating muscle flap for infected prostheses with deficient soft tissue coverage; and (3) salvage muscle flap for wound dehiscence or necrosis in the immediate postoperative period. These patients were observed for an average of 4.1 years (range, 1-8 years). The wound was revascularized successfully in 100% of the knees. The prosthesis was preserved in 83% of the knees. Recurrent infection occurred in 3 patients. Treating muscle flaps are least likely to result in an overall successful reconstruction because these flaps are used in knees with chronic infection. Prophylactic flaps and salvage muscle flaps provide the soft tissue coverage necessary to allow successful reconstruction in the appropriate circumstances. PMID- 7497659 TI - Knee arthrodesis in the treatment of failed total knee replacement. AB - Twenty-six primary arthrodeses and 6 repeat arthrodeses were done for a total of 32 arthrodesis procedures in 26 patients, with followup from 2 to 10 years (mean, 4 years). The indications for arthrodesis were septic failure in 18 patients and aseptic loosening in 8 patients. Thirteen arthrodeses were done using external fixators, and 12 arthrodeses were done using a closed fluted intramedullary nail for fixation (1 arthrodesis was done with a custom proximal and distal interlocked nail). In addition, 6 repeat arthrodeses with intramedullary nail fixation were done for treatment of nonunions after external fixation. Five (38%) of the 13 patients who underwent arthrodesis with an external fixator had clinical and radiographic union at a mean of 5 months. All 13 patients with primary intramedullary nail arthrodesis achieved union. The knees of the 6 patients with nonunions after external fixation that were treated with repeat intramedullary nail arthrodesis achieved union. Patients with septic failure had staged debridements before intramedullary nail arthrodesis. Intramedullary nail arthrodesis can be done safely in patients with sepsis as a staged procedure. Knee arthrodesis using intramedullary nail fixation gives a much higher union rate than does external fixation and is associated with fewer complications. PMID- 7497658 TI - Evolution of the quadriceps snip. AB - Between May 1990 and December 1992, the quadriceps snip was used to assist in the surgical exposure of the knee in 16 patients (4 women and 12 men) who underwent knee surgery at the average age of 65 years (range, 50-73 years). The original diagnoses included osteoarthritis (12 patients), traumatic arthritis (2 patients), rheumatoid arthritis (1 patient), and psoriatic arthritis (1 patient). The surgical exposure was done in the treatment of aseptic loosening of a total knee prosthesis in 8 patients, septic loosening in 2 patients, and primary knee arthroplasty in 6 patients with fibrous ankylosis. Using the Hospital for Special Surgery Scoring System, the authors rated the knees of 10 patients as excellent and those of 6 patients as good at postoperative evaluation. The range of motion was improved in all knees by an average of 30 degrees. Cybex testing revealed significant weakness of the knee in extension when compared with the contralateral normal side at test speeds of 60 degrees and 180 degrees per second but no difference compared with the opposite knee that had been replaced. In the authors' experience, the surgical technique of a quadriceps snip has been safe and simple, has not required special equipment, and postoperatively the patient's physical therapy has not had to be altered. Clinically, the quadriceps strength was very good, although weaker than that of a contralateral normal leg when measured by Cybex testing. PMID- 7497660 TI - History of the contralateral knee after primary knee arthroplasty for osteoarthritis. AB - Published reports of series of total knee replacements vary between 20% and 75% in the percentage of patients eventually having bilateral replacements. There are no data in the literature on the predicted course of the contralateral knee for patients presenting for total knee replacement. This study reviewed the history of the contralateral knee in patients with a diagnosis of osteoarthritis presenting for unilateral primary total knee arthroplasty. The current study group comprised 185 patients who had either a minimum 5 year followup (range 5 12.5 years) or who were known to have had a second knee arthroplasty before 5 years. Evaluation included a full clinical and radiographic examination of the contralateral knee at index arthroplasty and at final followup. Of the 185 patients, the contralateral side initially was asymptomatic in 36%, mildly symptomatic in 16%, moderately symptomatic in 28%, and severely symptomatic in 20%. Seventy-nine (43%) knees eventually underwent contralateral arthroplasty. Ninety-three percent of patients who had moderate or severe symptoms and severe radiographic arthritis of the contralateral side at the time of presentation later underwent total knee replacement. However, patients who presented initially with mild symptoms or who had no symptoms had only a 9% incidence of knee arthroplasty. PMID- 7497661 TI - Tibial wedge augmentation for bone deficiency in total knee arthroplasty. A followup study. AB - This report constitutes a followup of a previous study of 28 knees in 25 patients with metal wedge augmentation for tibial bone deficiency. Those patients were reviewed 2.3 years after surgery; 79% had excellent results, and 21% had good results. Twenty-four knees in 21 patients with a metal wedge augmentation for tibial bone deficiency were reviewed 5.6 years clinically and 4.8 years radiographically after surgery. Clinical results were excellent in 67%, good in 29%, and poor in 4%. The only poor result was in 1 knee that required 2 revision procedures: the first for failure of a metal-backed patellar component and the second for aseptic loosening of the femoral component. Radiolucent lines at the cement bone interface beneath the metal wedge were present in 13 knees. Eleven of those radiolucencies were < 1 mm in width, and 2 were 1 to 3 mm in width. None of those lucent lines were progressive. Metal wedge augmentation for tibial bone deficiency is a useful option. No deterioration of the wedge-prosthesis or wedge cement-bone interface was seen at midterm followup. PMID- 7497662 TI - Resurfacing versus not resurfacing the patella during total knee replacement. AB - One hundred patients with osteoarthritic knees were randomized either to have their patella resurfaced or not resurfaced using the same total knee replacement. These patients were assessed preoperatively and a minimum 2 years postoperatively using disease-specific (Knee Society Clinical Rating System) and functional capacity (30 second stair climbing and knee flexor and extensor torques) outcome measures. Two patients in the not resurfaced group required reoperation because of anterior knee pain. At 2 years' followup, the not resurfaced group had significantly less pain and better knee flexor torques than did the resurfaced group, whereas the results of the Knee Society Function Scores, 30 second stair climbing, and knee extensor torques were similar. These results suggest that longer-term followup is required, but that one should keep an open mind regarding patellar resurfacing during total knee replacement. PMID- 7497665 TI - Screw osteolysis after cementless total knee replacement. AB - A comparative radiographic review of 217 cementless total knee replacements was done 4 years after surgery to look for radiographic change at the screw-bone interface. There were 102 Miller Galante I knee prostheses, 59 Miller Galante II prostheses, and 56 Anatomic Modular Knee prostheses studied. If the appearance of the screw-bone interface had changed, this was classified according to the width of the lucency. Two hundred sixty-five of the 851 screws that were studied showed some evidence of change, with 185 (21.7%) of the screws showing a cavitary appearance change. The AMK prosthesis showed the higher incidence of cystic and cavitary changes. Polyethylene debris is thought to be responsible for generation of osteolysis and the radiographic appearance change. Design of the components and particularly the polyethylene attachment mechanisms may be responsible for the differences in the frequencies of osteolysis. PMID- 7497664 TI - The anteroposterior axis for femoral rotational alignment in valgus total knee arthroplasty. AB - This study evaluated a technique using the anteroposterior axis of the distal femur, rather than the transepicondylar or posterior femoral condylar axis, to establish rotational alignment of the femoral component in valgus knees. The anteroposterior axis of the distal femur was defined by a line through the deepest part of the patellar groove anteriorly and the center of the intercondylar notch posteriorly. Total knee arthroplasty was done in 46 valgus knees between 1980 and 1986 using the posterior femoral condyles as landmarks for rotational alignment. From January 1986 through January 1992 total knee arthroplasty was done in 107 valgus knees using the anteroposterior axis for rotational alignment of the femoral component. In the group of knees using the posterior condylar axis, medial tibial tubercle transfer was needed intra operatively in 8 knees to prevent lateral dislocation of the patella. In the first 2 postoperative years, 4 knees had recurrent patellar dislocation or subluxation that required surgical correction. In the group of knees using the anteroposterior axis, patellar tracking problems that required realignment were significantly reduced. One knee required medial tibial tubercle transfer to correct a Q angle > 20 degrees. In the remaining knees, the Q angle was < 10 degrees, and patellar tracking was acceptable. Two years after surgery, no knees had patellar instability. PMID- 7497663 TI - Is there a place for patellofemoral arthroplasty? AB - Indications for patellofemoral arthroplasty were studied based on a 2- to 10-year followup review. Of 183 patellofemoral prostheses implanted between 1980 and 1990, 104 were associated with unicompartmental arthroplasty, and 79 were implanted alone. Thirteen patients were lost to followup. The results of 66 patellofemoral arthroplasties are reported at an average followup of 66 months. The mean age of the patients at surgery was 57 years. Underlying causes of the osteoarthritis included patellar dislocation or dysplasia (22), patellar fracture (20), and primary osteoarthritis (24). A metallic femoral groove was associated with a polyethylene patella with no metal backing. Two (9%) prostheses were revised in the dysplasia group, 1 (5%) in the posttraumatic group, and 7 (29%) in the primary osteoarthritis group. In this latter group, the indications of patellofemoral arthroplasty must be restricted to lesions of the patellofemoral joint in a knee in neutral axis evaluated preoperatively on full standing and stress radiographs. The best results of patellofemoral arthroplasty were obtained in osteoarthritis after patellar dislocation or patellar fracture. PMID- 7497666 TI - Nonoperative treatment of rotator cuff tears. AB - In this study, 33 patients with full-thickness tears of the rotator cuff were given a supervised nonoperative program of rotator strengthening exercises and then were seen at followup an average of 3.8 years later. Of the 14 patients who were dissatisfied with nonoperative care, 12 eventually had opted for surgery and 2 were dissatisfied with their status at followup. In attempting to identify patients who would benefit from nonoperative care, the authors were unable to show that individual parameters such as rotator strength, symptom duration, or functional impairment were useful to predict outcome. However, it was found that those who have an insurance claim or who experience significant sleep loss due to shoulder pain are unlikely to be satisfied with nonoperative treatment. Patient satisfaction with nonoperative treatment is best correlated with improved pain relief, the ability to carry a 10- to 15-pound suitcase at one's side, the ability to use the arm at shoulder level, and the ability to eat using a utensil. Objective variables such as rotator strength and active motion are not correlated with ultimate patient satisfaction. PMID- 7497667 TI - Localized glenoid hypoplasia. An anatomic variation of possible clinical significance. AB - Examinations were done on 1150 scapular bone specimens from museum collections. A localized hypoplasia of the posteroinferior glenoid was found in significant numbers (20%-35%) depending on the population group studied. Criteria were established for the identification of this variant in computed tomography and magnetic resonance studies. Localized hypoplasia of the posteroinferior glenoid was detected as an incidental finding in 54 (18%) of 300 such studies. Using these same imaging criteria in a prospective manner, 9 of 12 patients seen for voluntary multidirectional instability of the shoulder displayed a localized posteroinferior glenoid hypoplasia. This phenomenon also may add to the picture of extreme posterior glenoid wear that sometimes is seen in association with osteoarthritis of the shoulder. PMID- 7497668 TI - Efficacy and safety of enoxaparin versus unfractionated heparin for prevention of deep venous thrombosis after elective knee arthroplasty. Enoxaparin Clinical Trial Group. AB - This study was a randomized, parallel-group, open-label clinical trial comparing the efficacy and safety of Enoxaparin, a low-molecular-weight heparin, and unfractionated heparin to prevent deep venous thrombosis after elective total knee arthroplasty. Four hundred fifty-three patients were randomized and received study medications. The primary efficacy evaluation was unilateral contrast venography done at the end of study or earlier if clinically indicated. The primary safety outcome was the incidence of bleeding episodes. Patients were assigned to 1 of 2 postoperative treatment groups: Enoxaparin 30 mg subcutaneous every 12 hours (228 patients), or unfractionated heparin 5000 units subcutaneous every 8 hours (225 patients). The incidence of proximal and distal deep venous thrombosis in the Enoxaparin group was 24.6% (56/228), and in the heparin group 34.2% (77/225). Three major hemorrhagic episodes were observed in each treatment group. Two cases of pulmonary embolism occurred in patients receiving heparin (1 fatal); no cases occurred in patients receiving Enoxaparin. This study showed that Enoxaparin administered postoperatively 30 mg every 12 hours is more effective and as safe as unfractionated heparin prophylaxis to prevent deep venous thrombosis in patients having elective total knee arthroplasty. PMID- 7497671 TI - Continuous passive motion after total knee arthroplasty. Analysis of cost and benefits. AB - The authors report the results of a prospective study examining the benefits of daily continuous passive motion combined with physical therapy, compared with physical therapy alone, in 103 consecutive osteoarthritic patients undergoing primary total knee arthroplasty. The first 51 patients received continuous passive motion initiated in the recovery room and the next 52 patients did not receive continuous passive motion. Both groups underwent an identical physical therapy protocol starting on the first postoperative day. At discharge, there was a significant increase in active flexion in the continuous passive motion group. There were no significant differences regarding pain, wound healing, knee swelling, wound drainage, pulmonary embolism, or length of hospital stay between the 2 groups. At 2 years, there were no clinical differences in the motion or knee scores. Knee manipulation was done for < 50 degrees flexion after the tenth postoperative day. There were 5 manipulations in the noncontinuous passive motion group and none in the continuous passive motion group. The entire costs associated with the 5 manipulations was $48,274 or $937 per patient not receiving continuous passive motion. The average daily inpatient rental of the machine was $60 per day. Continuous passive motion is efficacious in increasing short-term flexion and decreasing the need for knee manipulation without increasing costs. PMID- 7497669 TI - Extracorporal shock wave therapy for calcifying tendinitis of the shoulder. AB - During the past 2 years, 40 patients referred to the authors' hospital for persistent calcifying tendinitis of the shoulder were seen on prospective followup after undergoing a single extracorporal shock wave therapy. During a single therapy session, all patients received 1500 impulses of the energy density 0.28 mJ/mm2 in plexus anesthesia. Followup examinations were done at 6 and 24 weeks. In 62.5% of the patients partial or complete disintegration of the deposit was observed. Statistical analysis showed significant improvement both of subjective and objective criteria. According to the Constant score, 60% of the patients reached normal values, and 72.5% of the patients had no or only occasional discomfort. Only 6 patients (15%) reported no improvement at the 24 week followup. PMID- 7497670 TI - One-stage revision of total hip arthroplasty for deep infection. Long-term followup. AB - One hundred eighty-three total hip arthroplasties were revised for deep infection in a 1-stage procedure using antibiotic containing acrylic cement and systemic perioperative antibiotics. At an average followup of 7 years 9 months, 154 (84.2%) patients were free of infection. The remaining 29 (15.8%) patients had evidence of persistent infection. The quality of pain relief immediately after a revision for deep infection often is a good predictor of success. PMID- 7497672 TI - Secure fixation of lumbar osteotomy. Surgical experience with 50 patients. AB - Despite intensive conservative treatment, severe kyphotic spinal deformities may develop in patients with ankylosing spondylitis. Lumbar osteotomy can restore a horizontal visual field in these patients. Of 50 patients undergoing corrective lumbar osteotomy, there were 44 men and 6 women. A transpedicular screw and cable system was used in 37 patients. Mean correction of deformity was 38.7 degrees. Mean loss of correction was 4.8 degrees, but was 15 degrees in those cases in which transpedicular fixation was not used. All patients were able to see straight ahead after the operation. There were 2 deaths and 1 permanent sacral root lesion. Fixation failure was seen in 2 of 37 patients in whom transpedicular implants were used and in 3 of 13 in whom alternative methods of internal fixation were used. PMID- 7497674 TI - Salmonella panama osteomyelitis in an otherwise healthy patient. A case report. AB - A 22-year-old man had Salmonella panama osteomyelitis of the left distal tibia. He had endured a period of untreated diarrhea without fever 6 years before. The osteomyelitis was treated successfully with surgical debridement followed by 9 weeks of oral cotrimoxazole 960 mg twice daily. Salmonella osteomyelitis is rare. Most cases occur in patients with sickle cell anemia. Other conditions of local or generalized immunosuppression are also risk factors, but none were established in this patient, nor was he a chronic carrier. In reviewing the literature, no case of Salmonella panama osteomyelitis in an otherwise healthy patient was found. Although the osteomyelitis in this patient was possibly secondary to Salmonella enteritis 6 years before, the authors believe that enteric Salmonella infections should not be treated with antibiotics unless the infection is accompanied by systemic symptoms. Otherwise, the risk of chronic carriership is substantially increased. In case of Salmonella panama osteomyelitis, surgical debridement is recommended as the main component of treatment, followed by a prolonged period of specific antibiotic therapy. PMID- 7497673 TI - Tibial nonunion treated with direct current, capacitive coupling, or bone graft. AB - Two hundred seventy-one tibial nonunions of average duration of 23.5 months (range, 9-69 months) were treated with direct current (167 patients), capacitive coupled electrical stimulation (56 patients), or bone graft surgery (48 patients). Logistic regression analysis was used to compare heal rates among the 3 treatment methods, to identify risk factors adversely affecting the heal rate, and to predict the probability of successful healing of a nonunion of any given risk profile treated with each of the 3 forms of therapy. Seven risk factors were identified: duration of nonunion, prior bone graft surgery, prior electrical treatment, open fracture, osteomyelitis, comminuted or oblique fracture, and atrophic nonunion. When no risk factors were present, there were no significant differences among the 3 treatment methods. As progressively more risk factors were present, the predicted heal rates decreased significantly regardless of the treatment method. Some differences among the treatment groups did appear in the heal rates: bone graft surgery yielded a worse heal rate when there was a previous bone graft failure, and capacitive coupling had a worse heal rate in the presence of an atrophic nonunion. PMID- 7497675 TI - Treatment of cystic lesions of soft tissue using fibrin sealant. AB - A new method was devised for treating 29 patients with cystic lesions of the soft tissue (popliteal cysts, bursitis, and ganglia) by injecting a fibrin sealant into the cyst after the aspiration of the contents. During the followup period (mean, 2.2 years), cysts did not recur in 22 (76%) patients. Of the 7 patients with recurrences, 4 were treated again in the same manner and 3 of these cases have not recurred. Additionally, to investigate the healing process in cysts, fibrin sealant was injected into the space between the abdominal muscle and the peritoneum of rats. Specimens were obtained after the rats were sacrificed at 1, 2, 4, and 8 weeks after injection, and were examined histopathologically. The fibrin sealant was found to be completely resorbed and replaced with organized granulation tissue. In case of multiple failed aspirations to the cystic lesions, this new method is recommended before surgery. PMID- 7497676 TI - Giant cell tumor of bone. Curettage and cement reconstruction. AB - A retrospective review was conducted of all consecutive giant cell tumors treated by the senior author (JOJ) between 1975 and 1990 using the technique of aggressive curettage through a large bone window followed by acrylic cement reconstruction. Steinmann pins were used as reinforcement bars within the methacrylate in large lesions. Thirty-eight patients with a mean followup of 5.2 years (range, 2-16 years) were identified. Three (8%) recurrences are reported. There were no infections or mechanical failures of the construct. Functional results were good to excellent in 84% of patients. Ninety-five percent of patients reported good or excellent stability, no deformity, and emotional acceptance of the procedure. The recurrence rate with this protocol (8%) approached that associated with wide resection or amputation (0%-5%), and was lower than that seen with simple curettage and bone grafting (27%-55%). Acrylic cement reconstruction is a safe and effective procedure that provides local adjuvant therapy and immediate stability for early rehabilitation. The authors emphasize the importance of aggressive curettage of the lesion through a large bone window and suggest the use of reinforcement bars within the cement for large defects. PMID- 7497679 TI - Low back pain in a 23-year-old man. PMID- 7497678 TI - The Nicolas Andry Award. The pathomechanics and pathophysiology of cervical spinal cord injury. AB - Cervical cord injuries caused during American football games have resulted in reversible, incompletely reversible, and irreversible neurologic deficits. An explanation for this variable response to injury has been obtained from the study of the histochemical responses of a squid axon injury model to mechanical deformation. Data obtained indicate that recovery or lack thereof is directly proportional to the intracellular calcium concentration which in turn is directly proportional to the amount and rate of tension applied to the axon. It is concluded that in most instances of acute spinal injury, disruption of cord function is a result of the effects of local cord anoxia and the increased concentration of intracellular calcium. It is proposed that implementation of therapeutic measures that restore blood flow and reduce cytosolic calcium will increase neurologic recovery. PMID- 7497677 TI - Fatigue reduction by sequential stimulation of multiple motor points in a muscle. AB - The purpose of this study was to determine the optimal method of stimulating multiple motor points in a muscle in an attempt to improve fatigue resistance. The long head of triceps, which has 2 to 3 motor points, in an adult New Zealand white rabbit was used as the muscle model. The fatigue index, defined as the percentage of maximum force at time t, was compared during the fatigue tests. Five test groups were defined based on the stimulation pattern of the muscle: (1) Group A1: The proximal motor point was stimulated at 20 Hz; (2) Group A2: The distal motor point was stimulated at 20 Hz; (3) Group B: Both motor points were stimulated simultaneously at 20 Hz; (4) Group C: Both proximal and distal motor points were stimulated at alternate intervals of 10 seconds at 20 Hz; and (5) Group D: Both motor points were stimulated sequentially at 10 Hz. Each test was conducted for 6 minutes. In sequential stimulation (Group D), the fatigue index was significantly higher when compared with the other test groups at Minutes 4, 5, and 6. In groups A1, A2, B, and C, there were no significant differences in the fatigue indices. For optimal control of muscle contraction in functional electrical stimulation, electrodes should be inserted into all motor points for a given muscle. Sequential stimulation of these points can improve fatigue resistance. PMID- 7497680 TI - Magnetic resonance imaging of the musculoskeletal system. Part 1. Fundamental principles. AB - Magnetic resonance imaging has become a powerful tool for diagnosing many orthopaedic disorders. To use the technology appropriately and understand the nature of the images, it is important that the referring orthopaedic surgeon be familiar with the underlying principles of image formation. In this article, the author discusses the underlying physical phenomena, the instrumentation used for magnetic resonance imaging, and the principal determinants of the appearance of the image, in particular, the different pulse sequences. Paramagnetic contrast agents also are discussed briefly, and a few considerations regarding patient safety are addressed. PMID- 7497681 TI - Malrotation associated with implant alignment technique in total knee arthroplasty. AB - This study documents the malrotation between the femoral and tibial components associated with contemporary alignment techniques that position the tibial component relative to the tubercle, posterior tibial condyles, transtibial axis, and malleoli. The technique that allows the tibial component to float into alignment as the knee is passed through a range of motion and the technique of coupling the tibial component to the femoral component also were assessed. The average external rotation of the tibial component relative to the femoral component associated with each alignment technique is 19 degrees (tibial tubercle), 5 degrees (transtibial axis), 7 degrees (posttibial axis), 3 degrees (malleolar axis), and 14 degrees (range-of-motion technique). The coupled component technique produced 2 degrees internal rotation. The observed tendency to externally rotate the tibial component relative to the femoral component with most alignment techniques may account for the high incidence of posteromedial polyethylene wear reported in retrieval studies. PMID- 7497682 TI - Medial and anterior instability of the knee. An anatomical and clinical study using stress machines. 1971. PMID- 7497683 TI - Exposure in difficult total knee arthroplasty using tibial tubercle osteotomy. AB - Exposure with an extended tibial tubercle and tibial crest osteotomy was done for 136 total knee arthroplasties from 1986 to 1994: There were 26 primary arthroplasties, 76 revision, 10 repeated revision, 19 infected, and 5 repeated revision for infection. Adequate exposure was achieved and further release of the quadriceps mechanism was not necessary. Two or 3 wires were passed through the lateral edge of the tibial tubercle and through the medial tibial cortex to reattach the bone fragment and patellar tendon. Mean range of motion in these cases at 2 years after surgery was 93.7 degrees (range, 15 degrees-140 degrees). Two knees had extension lag, unchanged from their preoperative condition. Two tibial tubercles had partial proximal avulsion fracture, but did not separate widely. No evidence of non-union occurred in the simple cases or in the infected cases in which repeat elevation of the tibial tubercle flap and quadriceps mechanism was done. Three wires were removed because of pain. Two tibial fractures occurred in a single patient with diabetic Charcot arthropathy, and in 1 with manipulation after open adhesiolysis. Quadriceps function was not compromised in any case. Knees with Charcot arthropathy may need prolonged protection from weightbearing. Special caution should be exercised when manipulation is done to improve knee flexibility. PMID- 7497684 TI - Analysis of hospital cost in total knee arthroplasty. Does length of stay matter? AB - In this study, actual hospital total joint expenditures were analyzed, and the effect of decreased length of stay on these actual costs was determined. Expenditures were stratified per hospital and accounting conventions into fixed and variable costs. Hospital costs associated with 15 index knee arthroplasty procedures from fiscal years 1992, 1993, and 1994 were analyzed. Expenditures were standardized to inflation-controlled 1994 dollars. Average length of stay for knee arthroplasty procedures decreased 30% during this period (range, 8.7-6.1 days). Also, expenditures for total knee arthroplasty decreased only 13% (range, $17,415-$15,200) in inflation-controlled 1994 dollars. Using inflation-controlled dollars, analysis of variable costs revealed a continued rise of $278 during this period. Therefore, essentially all the cost reductions achieved were in the fixed component of cost allocated to each patient. Length of stay reductions, although beneficial, did not reduce hospital variable expenses significantly. Expenditure reductions were most exclusively in fixed costs. However, the overall total fixed hospital costs were largely unchanged. Thus, it is necessary to decrease variable costs (implant and supply costs) to significantly reduce hospital expenditures. PMID- 7497685 TI - Treatment of infected total knee arthroplasty using an articulating spacer. AB - Twenty-six patients with late infected total knee arthroplasties were treated by debridement and removal of components and all cement, preserving collateral ligaments. At time of debridement, an articulating spacer was fashioned to allow partial weightbearing and knee range of motion (ROM) during rehabilitation. This spacer was implanted using antibiotic-impregnated bone cement. For this purpose, 4.8 g of powdered tobramycin was mixed with each 40-g batch of Simplex cement. Cement was applied early to the components, but applied late to the femur, tibia, and patella to allow molding to the defects and bone without adherence to bone. Patients received tailored intravenous antibiotic therapy for 6 weeks in addition to this antibiotic-impregnated cement for treatment of a variety of gram positive and gram negative organisms. All patients had cemented revision total knee arthroplasty using antibiotic-impregnated cement with standard cementing techniques used. All patients but 1 had reimplantation; this patient died of unrelated causes before revision. Range of motion before revision was 10 degrees to 95 degrees. Followup averaged 30 months (range, 13-70 months). The average Modified Hospital for Special Surgery Knee Score after revision was 87 points (range, 53-100 points), with 92% good to excellent results. Range of motion after reimplantation was 5 degrees to 106 degrees. There have been no recurrences of infection. Use of an articulating spacer to treat infected total knee arthroplasty improves ultimate ROM and soft tissue health and significantly decreases the risk of reinfection. PMID- 7497686 TI - Patient outcome with reinfection following reimplantation for the infected total knee arthroplasty. AB - From 1976 to 1992, 24 knees (24 patients) were treated for reinfection after reimplantation of a prosthesis to treat an infected total knee arthroplasty. Including the index total knee arthroplasty, the average number of total procedures for the affected knee was 9.3 procedures (range, 5-23 procedures), and surgical procedures used for the affected knee after reinfection averaged 3.7 procedures (range, 1-12 procedures). The final outcome included 10 knees with a successful knee arthrodeses, 5 patients with infected prostheses who were prescribed suppressive oral antibiotic treatment, 4 above the knee amputations, 3 persistent pseudarthroses of the knee joint, 1 resection arthroplasty, and 1 uninfected total knee prosthesis. A poor prognosis was associated with use of a hinged knee design: 3 of the 4 amputations were done after a hinge knee prosthesis was implanted; 2 patients had a persistent pseudarthrosis after a failed attempt at knee arthrodesis; 1 patient had a resection arthroplasty; and 1 patient had a retained infected prosthesis. A successful arthrodesis in which the initial attempt included use of an external fixation devices was more likely for prostheses without stems (75%) when compared with cemented stemmed prostheses (40%). Long intramedullary arthrodesis was successful in all 3 attempts. Aspiration for diagnosis followed by oral antibiotic suppression was universally unsuccessful in 4 patients, whereas early aggressive open debridement facilitated retention of the prosthesis in 4 of 6 patients. The increased difficulty in achieving a healed wound, a successful knee arthrodesis, and eradication of infection with subsequent nonprosthetic salvage procedures as well as the attendant morbidity associated with reinfection must be considered carefully and discussed with the patient before the reimplantation prosthesis is inserted to treat the infected total knee arthroplasty. PMID- 7497687 TI - Long-term changes in bone mineral density following total knee replacement. AB - Bone density was measured using dual photon absorptiometry and dual xray absorptiometry below the tibial component under the medial and lateral plateaus, under the central peg, and distal to the central peg. Measurements of bone density were taken at 1 week, 6 weeks, 6 months, and 1 year after surgery in 31 patients. Seven of these patients also had measurements taken 8 years after surgery. The average age of the patients at surgery was 69 years old. One week after surgery, the mean bone density from all regions was 0.77 g/cm2 in women compared with 1.14 g/cm2 in men. Between 6 weeks and 6 months, the bone density below the tibial component declined inconsistently in most patients, but by 1 year the bone mineral density was statistically equal to the initial bone density at 1 week after surgery. Eight years after total knee arthroplasty, there was a 36.4% decrease in bone mineral density between men and women was not statistically significant. Medial plateau, lateral plateau, under the peg, and distal to the peg bone density changes tended to be the same. Maximum rates of bone resorption in patients with osteoporosis of the same age group have been reported to be between 1% and 2% per year. In this study, after an initial stable period, the bone density of the proximal tibia beneath the tibial component consistently decreased at a rate of 5% per year. Surgeons must consider this relatively large decrease in bone density if they are relying on bone for long term stable fixation of the prosthesis.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7497688 TI - Radiographic characterization of aseptically loosened cementless total knee replacement. AB - From January 1980 to December 1992, 655 cementless total knee arthroplasties with the PCA total knee system were done at the authors' institution. Thirty of these knees have been revised for aseptic loosening of either tibial or femoral components or both. Patients in the failed arthroplasty group were matched directly to a control group with well-functioning, stable and painless cementless total knee prostheses (Knee Society objective score, > 90 points). In this study, the authors analyzed any alignment differences between the 2 groups of knees. Special attention was given to the postoperative radiographs to evaluate for potential predictors of success versus failure. Multiple radiographic parameters were measured for each knee and included measures of frontal alignment, sagittal alignment, rotation, percent prosthetic coverage, and cortical thickness (denoted as the cortical-cancellous index). On univariate analysis, no individual radiographic parameter was significantly different in the failure group versus the control group. However, multivariate analysis indicated that certain combinations of abnormal parameters were significantly higher in the failed group than the control group. These parameters included measures of frontal and sagittal plane component alignment. The summated abnormal parameters per patient averaged 3.67 (range, 1-7) for the failure group and 1.80 (range, 0-7) for the control group. PMID- 7497689 TI - Flat-on-flat, nonconstrained, compression molded polyethylene total knee replacement. AB - Flat-on-flat, posterior cruciate ligament-sparing total knee prostheses recently have shown problems of wear, loosening, and multiple design changes. Two thousand one Anatomical Graduated Components total knee arthroplasties with compression molded, nonmodular polyethylene tibial components were done between 1983 and 1991 at 3 institutions. All knees were evaluated clinically and radiographically every 2 to 3 years; 71 knees were seen in followup > 10 years. There were 8 failures secondary to revision (5 tibial failures; 2 secondary to metalosis from patellar polyethylene dissociation; and 3 femoral failures) resulting in a 98% survival rate at 10 years. The tibial design was flat-on-flat with a compression molded polyethylene that the authors believe is the primary reason for its success. PMID- 7497690 TI - Effect of component fixation method on osteolysis in total knee arthroplasty. AB - Eighty-three total knee arthroplasties done at a single university hospital were reviewed specifically to examine the presence of lysis. Components that were radiographically loose were excluded. The incidence of lysis varied significantly with the method of component fixation. The highest incidence of lysis (30%) was seen when the tibial component was fixed with cement and screws and the femoral component was implanted without cement. When the tibial component was fixed with cement and screws and the femoral components was cemented, the incidence of lysis was 13% (2 of 13). When the femoral component was press fit and the tibia was cemented without screws, the incidence of lysis was 10% (1 of 10). When the femoral and tibial components were cemented and no screws were used, the incidence of lysis was 0 (0 of 12). Using screws with cement to fix the tibial component was associated with a high incidence of lysis and cannot be recommended. A press fit femoral component also may contribute to the incidence of lysis. PMID- 7497692 TI - Catastrophic osteolysis in total knee replacement. A report of 17 cases. AB - One hundred eighty-five total knee revisions were done by the 2 senior authors from 1988 to 1994. Of these, 17 patients were identified as having severe osteolysis. The average age was 62.7 years, and the average weight was 90.8 g. All patients had osteoarthritis. Preoperative aspiration was negative in all patients. Eight patients had a metal-backed patella, 8 had a cementless implant, 4 had a cemented implant, and 5 had a hybrid implant. The average time interval from the index surgery to radiographic evidence of lysis was 56 months. Radiographic review showed gross polyethylene wear in 10 patients, loose tibial implants in 8, fractured baseplates in 2, and evidence of a metallic synovitis in 2. Retrieved polyethylene liners had a average thickness of 7 mm, and none were conforming. Osteolytic defects were defined and classified according to Engh's criteria. The histology showed a foreign body reaction with intracellular refractile polyethylene particles. Four cases had metallic debris. The exposure at the time of revision required a rectus snip in 4, V-Y-quadricepsplasty in 2, and tibial tubercle osteotomy in 1. Posterior stabilized implants were used in 65% and a constrained implant in 30%. Lytic defects were reconstructed with cement only in 47% of knees, allograft in 30%, and metallic wedges in 35%. Based on this report, the authors conclude that younger, overweight patients seem to be at higher risk of implant failure and that osteolysis is not restricted to cementless implants. Furthermore, because review of the radiographs leads to a constant underestimation of the lytic defect, the surgeon must be prepared to deal with complex revisions. PMID- 7497691 TI - Effect of porous-coating configuration on tibial osteolysis after total knee arthroplasty. AB - Porous-coating configuration on the undersurface of tibial components in total knee arthroplasty was evaluated for its effect on tibial osteolysis. Clinical data and radiographic results of 675 Ortholoc II components (range, 4-8 years postoperative) and 236 Ortholoc Modular components (range, 2-4 years postoperative) were compared. The undersurface of the Ortholoc II tibial component is completely covered with porous coating, and that of the Ortholoc Modular is partially porous coated with smooth metal bridges connecting the 4 screw holes and joint cavity to the smooth metal stem. None of the radiographs of the Ortholoc II tibial components showed significant osteolysis around the stem or pegs. Radiographically detectable signs of osteolysis around the distal 1/2 of the stem appeared in 28 of the 124 long-stemmed Ortholoc Modular knees. None had radiographically identifiable osteolysis around the screws or pegs. In the 112 knees with short stems, 19 had radiographically identifiable radiolucency around the stem, but none had radiolucency around the screws or pegs. Despite their substantially shorter followup, cementless tibial components with patches of porous coating connected by smooth metal tracks were more likely to have osteolysis develop than those with uninterrupted porous coating on the undersurface of the tibial tray. The smooth metal tracks appear to conduct debris from the joint cavity to the areas surrounding the stem. PMID- 7497693 TI - Patterns of individual diet choice and efficiency of foraging in wedge-capped capuchin monkeys (Cebus olivaceus). AB - Capuchin monkeys (Cebus olivaceus) exhibit extensive intragroup variability in foraging and diet. To consider how age, sex, and individual identity contribute to this variability, the authors examined foraging and diet in 18 wedge-capped capuchin monkeys in 1 social group in the wild. Age-sex classes did not differ in the time spent ingesting food, the reliance on plant foods, the foraging actions used or substrates exploited, or in the efficiency of exploiting animal foods. They did differ, however, in the time spent finding food, time devoted to animal foods and to vigorous foraging, and the efficiency of foraging. The sexes differed more than age groups. Individual differences within age-sex class were less extensive than expected and were more evident in juveniles than adults. Within-group variability in foraging reflected catholic selection and equivalent treatment of substrates by all individuals, rather than individual specializations. PMID- 7497694 TI - Scent counter-marks: specialized mechanisms of perception and response to individual odors in golden hamsters (Mesocricetus auratus). AB - Scent counter-marking is a wide-spread phenomenon among mammals that is not well understood. Using a habituation paradigm, it was found that male golden hamsters (Mesocricetus auratus) exposed to partially overlapping scents of 2 females behaved on test trials as if they remembered the top scent but not the bottom scent. If the 2 females' scents did not overlap during exposure trials, males remembered both of them. Thus, if 2 individual scents partially overlap, hamsters (a) perceive which of the 2 scents was deposited most recently and (b) either remember the top scent and forget the bottom scent or attach greater significance to the top scent. These results indicate the existence of specialized perceptual and cognitive or motivational mechanisms that are used for interpretation and response to complex arrays of social odors. PMID- 7497695 TI - Behavioral responses of longtailed macaques to different cage sizes and common laboratory experiences. AB - The authors tested the effects of varying cage size on the behavior of 10 female and 10 male Macaca fascicularis by singly caging them for 2 weeks in each of 5 cage sizes, ranging from approximately 20% to 148% of regulation size. Behavior in the regulation cage size, a size 23% smaller, and a size 48% larger did not differ in any analysis. Locomotion was significantly less in the 2 smallest cage sizes. Abnormal behavior occurred only 5% of the time, did not increase as cage size decreased, and did not change significantly over nearly 3 years. Disruption of the normal activity budget in the laboratory environment proved to be a useful indicator of psychological well-being. Moving to a new room and, to a lesser extent, moving into a new, clean cage, regardless of size, was associated with disrupted sleep the 1st night and suppressed activity, especially self-grooming, the next day. PMID- 7497696 TI - Diet preference in rats (Rattus norvegicus) as a function of odor exposure, odor concentration, and conspecific presence. AB - The relative contributions of social and stimulus factors in development of rat dietary preferences were examined. Investigation of odor-alone effects revealed that weak odors resulted in preference for familiar-odor diets, but only at longer exposure times. Shorter exposure to strong odors also produced differences in diet preference. When odor and conspecific presence were manipulated simultaneously, odors produced no diet preference at low intensities, whereas high-intensity odors did so regardless of conspecific presence. Medium-intensity odor concentrations produced differences only with conspecifics present, indicating social enhancement of stimuli that are ineffective in isolation. These results suggest the separate influence of social and stimulus factors on dietary preferences and explain contradictions in previous studies. PMID- 7497697 TI - Modes of vocal variation in Sykes's monkey (Cercopithecus albogularis) squeals. AB - Acoustic and laryngographic measurements of Sykes's monkey (Cercopithecus albogularis) squeals showed that acoustic variation between exemplars was principally due to 4 underlying modes of modification: (Class I) variation in the transfer function produced by articulation, (Class II) variation in F0 produced by inflection, (Class III) variation in turbulence, and (Class IV) variation in the pattern of vibration of laryngeal tissue with large or small surface areas (polyphonic variation). These findings suggest that some modes of variation (Class I, II, and III calls) were due to phonatory and articulatory acts resembling, at a very general level, sound production mechanics used in speech. Acoustic variation in the polyphonic group (Class IV calls) is not evident in speech and was attributed to the possibility of independent and simultaneous vibrations of the vocal lip and the principal component of the vocal folds. PMID- 7497698 TI - Assessment of danger to themselves and their infants by rhesus macaque (Macaca mulatta) mothers. AB - This study investigated whether rhesus macaque (Macaca mulatta) mothers distinguish between what is dangerous to themselves and what is dangerous to their infants. The behavioral interactions between 11 mother-infant pairs and other females living in their group were analyzed in the 1st 2 months of infant life. Mothers behaved as if they perceived higher ranking females as dangerous to both themselves and their infants, lower ranking females as dangerous to their infants but not to themselves, and their young daughters as relatively harmless to both themselves and their infants. Changes in maternal intolerance of infant handling between the 1st and 2nd months covaried with changes in the probability of infant harassment rather than with the temporal pattern of aggression received by mothers. The possible cognitive mechanisms underlying parental recognition of infant's special needs are discussed in the light of comparative evidence from other mammalian species. PMID- 7497699 TI - Multiresolution decomposition of non-stationary EEG signals: a preliminary study. AB - Wavelet representation is a recent development in the analysis of non-stationary signals. Its possibilities for use in the description of time-frequency characteristics of both transients in spontaneous EEG and time-varying rhythms in event related brain activity are explored here. By way of illustration, multiresolution decompositions of a wide variety of EEG transients are carried out in this work, including spike-and-waves, single spikes, sharp waves, blink artifacts, frontal intermittent rhythmic delta activity (FIRDA) and paroxysmal delta activity. Also, the application of the wavelet representation to study related spectra perturbations is illustrated with data from psychophysical experiments on the perception of image motion. The results demonstrate the capabilities of the wavelet transform, as an alternative to the Fourier transform, for the representation and analysis of non-stationary EEG signals. PMID- 7497700 TI - Comparison of FFT- and AR-based sonogram outputs of 20 MHz pulsed Doppler data in real time. AB - Real time sonogram outputs of autoregressive (AR) and Fast Fourier Transform (FFT) spectral analysis of 20 MHz pulsed ultrasonic Doppler blood flowmeter are presented. Data obtained from coronary, renal, iliac, digital and mesenteric arteries were processed using AR- and FFT-based spectral analysis techniques and interpretable sonograms were constructed. In comparison with the FFT-based sonogram outputs. AR-based sonogram outputs for 20 MHz pulsed Doppler data provide better results. Hence, the AR modeling is strongly recommended for small vessels with diameters between 1 and 2 mm. PMID- 7497701 TI - Medical image processing utilizing neural networks trained on a massively parallel computer. AB - While finding many applications in science, engineering, and medicine, artificial neural networks (ANNs) have typically been limited to small architectures. In this paper, we demonstrate how very large architecture neural networks can be trained for medical image processing utilizing a massively parallel, single instruction multiple data (SIMD) computer. The two- to three-orders of magnitude improvement in processing time attainable using a parallel computer makes it practical to train very large architecture ANNs. As an example we have trained several ANNs to demonstrate the tomographic reconstruction of 64 x 64 single photon emission computed tomography (SPECT) images from 64 planar views of the images. The potential for these large architecture ANNs lies in the fact that once the neural network is properly trained on the parallel computer the corresponding interconnection weight file can be loaded on a serial computer. Subsequently, relatively fast processing of all novel images can be performed on a PC or workstation. PMID- 7497702 TI - The assessment of LH surge for predicting ovulation time using clinical, hormonal, and ultrasonic indices in infertile women with an ensemble of neural networks. AB - An ensemble of independently trained neural networks (NN) is proposed for the assessment of luteinizing hormone (LH) surge for predicting ovulation time in infertile but ovulating women. The proposed ensemble involves a number of parallel NN modules. Each pair of the NNs learn specific data that are previously collected for monitoring timing function of LH estradiol (pg ml-1), and follicle diameter (mm) are used to train NN pairs to approximate the function of the LH values. A reasonable and accurate estimation places ovulation approximately 10-12 h after the LH peak. The double-valued (bi-phasic) regions of training data are separated into two single-valued (bi-phasic) regions of training data are separated into two single-valued parts (not exactly preovulatory, postovulatory division) that can be learned by each module of the NN pair. During testing, after the initial decision to have single-valued sides, the assessment is obtained by a linear opinion pool (consensus rule) using the decisions of NNs on the corresponding side without waiting. The network ensemble has various desirable properties: high assessment accuracy of a double-valued multisource data, minimized learning and recall times, and a parallel structure. The ovulation time can be predicted through the assessment of LH peak with a better precision and fewer number of tests. PMID- 7497703 TI - A C+ + class library foundation for developing an electronic medical record. AB - The basic features that gave the M (or Mumps, as it originally called) language its initial popularity are still important to many programming applications, especially those involving the health care domain. To date, however, efforts to change the language have been predicated on the axiom that the existing structure must be preserved in order to accommodate legacy applications. To many, it is becoming clear that the 1960s' minicomputer induced interpreter syntax underlying M is inadequate and incapable of further evolution. In this paper, we demonstrate an alternative that preserves those features that many find to be the essence of M in a manner that is transportable to other language environments and not directly linked to any underlying operating system. This article describes a set of operating system independent C + + class libraries, based on M, designed to enhance the C + + programming environment with features that facilitate health care record applications development. These extensions allow the user to manage hierarchical data base problems, such as those typical in the health care domain, at a level of abstraction consistent with many information theoretic models commonly in use. Examples are given from a pilot implementation of the libraries to demonstrate a simple health care record application. PMID- 7497704 TI - A PC program for unbiased and predictive linear and quadratic discriminant analysis. AB - Discriminant analysis plays an important role in biological and medical research. The most popular methods of discrimination in practical applications are parametric methods like linear and quadratic discriminant analysis. However, there exist modifications of these approaches, namely unbiased and predictive discriminant analysis, which lead to reduced error rates in certain situations. In this paper a menu-driven, user-friendly PC program written in Borland Pascal 7.0 is introduced which performs unbiased and predictive linear and quadratic discriminant analysis. PMID- 7497706 TI - Drug abuse and psychiatric comorbidity. AB - Psychiatric morbidity was studied in a sample of 176 opioid-dependent men recruited from prison and treatment services. The Diagnostic Interview Schedule (DIS) was used for psychiatric assessment. Lifetime and current prevalence of any mental disorder, excluding substance use disorders, reached 90.3% and 66.1%, respectively. The most prominent lifetime DSM-III axis I disorders were anxiety (31.8% lifetime and 16.5% last month) and affective (25% lifetime and 19.9% last month) disorders. Antisocial personality disorder (ASP) had a lifetime prevalence of 69.3%. Higher rates of affective and anxiety disorders were diagnosed in the treatment sample than in the imprisoned sample, whereas the inverse was observed for ASP. Results from the psychiatric interview showing a strong association between drug--opiate--dependence and DSM-III mental disorders are further supported by findings of high levels of depressive symptoms on the Center for Epidemiological Studies-Depression (CES-D) scale (71.5%) and increased rates of self-reported suicide attempts (27.4%) and psychiatric hospitalizations (26.8%). Psychiatric disorders seem to precede drug dependence in the majority of cases. Results from this study are compared and discussed with those from similar North American Studies. PMID- 7497707 TI - Descriptive validity and stability of diagnostic criteria for dysthymic disorder. AB - The goal of this study is to examine different criteria for the diagnosis of dysthymic disorder (DD). Using a two-stage design, records of 37 subjects diagnosed with DSM-III-R DD were systematically reviewed. Seven of these patients met criteria except for having depression less than 50% of the time. The initial evaluation involved a comparison of the diagnostic assessments using symptom criteria of DSM-III-R and ICD-10, proposed DSM-IV criteria for DD, DSM-III-R major depressive disorder (MD) criteria, and the Columbia criteria for atypical depression (AD). A subsample of 16 subjects who were assessed at least twice several months apart was then extracted. Agreement between the assessments with the proposed DSM-IV symptom list was determined. In the initial assessment, all 37 subjects (100%) met DSM-III-R and ICD-10 criteria, proposed DSM-IV criteria for DD, and DSM-III-R MD criteria, with percentages of mean positive items on the four criteria being 77%, 84%, 89%, and 77%, respectively. In addition, nine subjects (24.3%) also met diagnostic criteria for AD. All 16 subjects (100%) of the subsample again met proposed DSM-IV criteria for DD when assessed 75 to 145 days after their initial assessment. Items with the highest agreement between each two assessments were pessimism and low self-esteem, whereas social withdrawal and decreased activity had lower agreement. The proposed DSM-IV criteria for DD had good sensitivity and reliability in assessing core symptoms of DD. The use of different criteria for DD is discussed. PMID- 7497705 TI - Specificity of substance use in anxiety-disordered subjects. AB - The current research (1) examines empirical evidence to substantiate the relationship between substance choice and chronology of onset of anxiety and substance use disorders, and (2) provides information on the specificity of substance choice among anxiety disorders. A study group of 181 subjects in the Harvard Anxiety Research Project (HARP) who had a history of substance use disorder were the focus of this examination. Subjects whose anxiety disorder had an onset before their substance use disorder (primary anxiety) were compared with those whose substance use preceded onset of an anxiety disorder (secondary anxiety) for differences in distribution of subjects among categories of substance of abuse. Primary and secondary anxiety groups do not have different ages of onset for substance use disorder, nor was there greater likelihood for choosing alcohol for any of the anxiety disorders. However, there is a decreased risk of alcohol use in the small group of generalized anxiety subjects and an increased risk of opioid use in the small group of posttraumatic stress disorder subjects. There was no indirect support for the self-medication hypothesis. Neither age of onset data, specific substance association, nor proximal diagnosis association support a simple interaction. The strongest finding supported an "avoidance" of CNS stimulants. PMID- 7497708 TI - An attempt to validate Akiskal's classification of chronic depression using cluster analysis. AB - A numerical taxonomy program that overcomes several problems often associated with cluster analysis was used in an attempt to validate Akiskal's classification of chronic depression. Whereas two of the four groups produced by this technique demonstrated features of either Akiskal's character spectrum disorder or chronic secondary depression, one group appeared to subsume both Akiskal's chronic primary depression and subaffective dysthymia. Within this group, early-onset patients exhibited characteristics of subaffective dysthymia and late-onset patients demonstrated features of chronic primary depression. Family history of depression and family history of alcohol dependence were potent discriminators between groups. There were points of similarity between the typology and several classifications of episodic depression, for example, Winokur's classification of unipolar depression and other typologies derived from multivariate analysis. PMID- 7497709 TI - Are positive and negative symptoms relevant to cross-sectional diagnosis of schizophrenic and schizoaffective patients? AB - The value of positive and negative symptoms for cross-sectional differential diagnosis was studied in a sample of 103 consecutively admitted patients with schizophrenia and schizoaffective psychoses. A semistructured interview for schizophrenia was used. Subjects were diagnosed by Research Diagnostic Criteria (RDC) and classified as schizophrenic, depressive schizoaffective, and manic schizoaffective disorders. DSM-III-R criteria for schizophrenia were used to assign RDC schizophrenics to two groups: "true" schizophrenic (fulfilling both DSM-III-R and RDC criteria) and schizophreniform (fulfilling RDC but not DSM-III R criteria). The Scale for the Assessment of Positive Symptoms (SAPS) and Scale for the Assessment of Negative Symptoms (SANS) were used. Our findings showed that both positive and negative symptoms were relevant to differential diagnosis between schizophrenia and other psychotic disorders. However, negative symptoms presented higher significant differences between diagnostic groups than positive symptoms. These results were obtained by global ratings and by scoring on individual symptoms of SAPS and SANS. Treatment with neuroleptic and biperidene drugs did not alter symptomatologic differences between groups that resulted from repeated analyses of covariance. Certain symptoms and clusters of symptoms were found to be of higher severity of higher severity in only one diagnostic group. They were as follows: pressure of speech for manic schizoaffective disorder; guilt for depressive schizoaffective; high scores on negative symptoms for depressive schizoaffective and schizophrenic disorders; inappropriate affect for schizophrenic disorder; and three "psychomotor retardation" symptoms of the affective flattening subscale of SANS (unchanging facial expression, decreased spontaneous movements, and paucity of expressive gestures) for depressive schizoaffective disorder.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7497712 TI - Cultural challenges to the psychiatric enterprise. AB - Two challenges to psychiatry emanating from the study of cultural differences in mental illness are reviewed. The first challenge, conducted under the banner of cultural relativism, appeared to undermine the validity of psychiatric disorders. The misconceptions of this challenge, as embodied in the excesses of the labeling theory of sociology, were associated with the writings of antipsychiatrists and rejection of the community mental health movement. The result of all this was a devaluation of insights inherent in the cultural perspective. The second challenge draws emphasis to the pervasive role played by cultural differences in every facet of the psychiatric enterprise. Its relevance and contributions are briefly summarized. The study of the cultural sciences as they pertain to psychiatry offers a necessary corrective to the increasing impersonality and reductionism that is coming to characterize the neurobiologic approach. PMID- 7497710 TI - Identity disturbances and self-other differentiation in schizophrenics, borderlines, and normal controls. AB - The present study investigates identity disorders in schizophrenics and borderlines. Nineteen schizophrenics and 17 borderlines were compared with 18 normal controls. The technique used was an adapted version of the repertory grid test to describe the self and nine significant others (i.e., family members). Three indices were derived from the 10 person x 20 self-generated-attribute matrix to measure the extent to which self was differentiated from others: (1) overlap of salient attributes, (2) overlap of opposite attributes, and (3) degree of differentiation among others. Results showed that both schizophrenics and borderlines describe themselves more in terms of opposites than in terms of salient attributes. Differentiation among significant others was severely impaired in schizophrenics and preserved in borderlines. These findings were interpreted as a failure of the individuation process in schizophrenics and as an incomplete construal of self-identity in borderlines. PMID- 7497711 TI - Insight in clinical psychiatry revisited. AB - Current research on insight in psychiatry has focused on exploring the relationship between insight and other clinical variables such as severity of psychopathology, compliance with medication, general outcome, and, more recently, neuropsychological impairments. However, the studies yield divergent and inconsistent results, and consequently the role of insight in relation to such variables remains unclear. We review here recent studies in this area and suggest that one reason for variability in the results relates to confusion surrounding the term "insight." It is argued here that more conceptual work is needed to help clarify the meaning of insight before further empirical research is undertaken. Some of the conceptual issues are raised, and it is suggested that the concept of insight is first differentiated from the phenomenon of insight (its clinical representation) to facilitate a more structured and systematic approach to the examination of insight in clinical practice. PMID- 7497713 TI - Twelve-month test-retest reliability of the structured clinical interview for DSM III-R personality disorders in cocaine-dependent patients. AB - This study examined 12-month test-retest reliability of the Structured Clinical Interview for DSM-III-R Personality Disorders (SCID-II) in cocaine-dependent patients. Thirty-one patients completed the SCID-II during the second week of hospitalization for cocaine dependence, and again 12 months later. In both interviews, patients were asked to answer questions about their personality during the several years preceding admission to the hospital. Test-retest reliability, as measured by kappa, was relatively poor at .46. However, reliability of negative diagnoses (the absence of a disorder at both time points) was higher than reliability of positive diagnoses (the presence of a disorder at both time points). Reasons for the difficulty in attaining long-term test-retest reliability of axis II diagnoses in cocaine-dependent patients are discussed. PMID- 7497715 TI - Humoral tea leaves: what's at the bottom of the cup? PMID- 7497714 TI - A new approach to affective symptoms in relapsing-remitting multiple sclerosis. AB - A variety of psychiatric disorders accompany multiple sclerosis (MS), affecting the spheres of cognition, affect, and personality. These disorders may be of primary and/or secondary nature, reflecting, respectively, neurologic damage and/or functional distress associated with a disabling disease. Evaluation of the nature, severity, and frequency of affective symptoms of MS patients is of importance to sensitize the treating physician to the possible need for specific psychiatric treatment. We conducted a prospective cross-sectional study assessing correlations of psychiatric symptomatology and the neurologic parameters of disease duration, disease activity (as expressed by the number of disease exacerbations per year), and disease severity (as measured by the Kurtzke Expanded Disability Status Scale [EDSS]) in 20 relapsing-remitting (RR)-MS patients. Patients were also evaluated by a semistructured psychiatric interview and the following rating scales: Hamilton Anxiety Scale (HAS), Hamilton Depression Scale (HDS), Hackett-Cassem Denial Scale, and Lubin Adjective Affective List. Results demonstrated a high prevalence of anxiety (90%) and to a lesser extent depression (50%) in RR-MS patients. Depression and anxiety were found to be associated with disease activity, but not with disease duration or severity. The mechanism of denial played a significant role in the psychiatric profile of the disease and was correlated with disease duration. We suggest that in RR-MS patients, psychiatric evaluation should also include assessment of defense and coping mechanisms, which in turn could guide specific individualized treatment. PMID- 7497716 TI - Anti-interleukin-6 antibody treatment improves survival during gut-derived sepsis in a time-dependent manner by enhancing host defense. AB - OBJECTIVE: To determine the in vivo neutralizing activities of anti-interleukin-6 (IL-6) antibody on survival rate and host defense in a clinically relevant model of infection. DESIGN: Prospective, randomized, experimental animal study. SETTING: University and Shriners Burns Institute research laboratories. SUBJECTS: Two hundred seventy-six adult, female Balb/c mice. INTERVENTIONS: Balb/c mice were treated with 10 micrograms of antimurine IL-6 antibody, nonspecific murine immunoglobulin G (IgG), or placebo at 2, 4, or 8 hrs after they underwent bacterial challenge by gavage of 10(10) Escherichia coli and thermal injury. The survival rate was determined. The number of viable translocated bacteria, the total amount of translocation, and the percentage of bacteria that survived were also studied in different tissues. MEASUREMENTS AND MAIN RESULTS: Survival rate after burn and gavage was significantly improved in animals treated with antimurine IL-6 antibody at 2 and 4 hrs but not at 8 hrs after injury compared with control animals treated with nonspecific IgG or saline. The IL-6 serum concentration was significantly lower after burn and gavage in the animals treated 2 and 4 hrs after injury compared with nontreated animals. Better killing of translocated bacteria was observed in the tissues of animals treated with antimurine IL-6 antibody 2 hrs after injury. CONCLUSIONS: Treatment with antimurine IL-6 antibody at 2 and 4 hrs after injury, but not at 8 hrs after injury, positively affects outcome during gut-derived sepsis. Moreover, the beneficial effect of treatment after 2 hrs was related to an enhanced clearance of translocated bacteria. PMID- 7497719 TI - Early hemodynamic course of septic shock. AB - OBJECTIVE: To assess the relative contributions of changes in vascular tone and changes in cardiac function to hemodynamic recovery from septic shock. DESIGN: Case series, observational study. SETTING: Multidisciplinary department of intensive care in an academic hospital. PATIENTS: Sixty-seven patients with septic shock (prolonged hypotension, signs of tissue hypoperfusion, signs of sepsis, suspected source of infection, or documented bacteremia). In addition to the antibiotic therapy and the removal of the source of sepsis whenever possible, each patient received intravenous fluids and vasoactive agents (dopamine, norepinephrine, and dobutamine). Each patient was also treated with mechanical ventilation. Twenty-four (36%) patients survived their intensive care unit course. INTERVENTIONS: Hemodynamic measurements were obtained at baseline, after initial resuscitation (as soon as apparent hemodynamic stability was achieved), after 12 hrs, and after 24 hrs. MEASUREMENTS AND MAIN RESULTS: There were no significant differences in hemodynamic or oxygen-derived variables at baseline between the survivors and the nonsurvivors. During the initial resuscitation period, only the survivors demonstrated a significant increase in mean arterial pressure (from 69 +/- 17 to 82 +/- 18 mm Hg; p < .02) and left ventricular stroke work index (from 25.2 +/- 11.0 to 35.5 +/- 19.4 g.m/m2; p < .05). The increases in cardiac index and systemic vascular resistance were greater in the survivors than in the non-survivors, but the differences did not reach statistical significance. Study of the left ventricular function curves indicated an improvement of left ventricular function in the survivors but not in the nonsurvivors. CONCLUSION: An early improvement in left ventricular function is a hallmark of the survivors from septic shock. PMID- 7497718 TI - Comparison of dopamine to dobutamine and norepinephrine for oxygen delivery and uptake in septic shock. AB - OBJECTIVES: To test whether dopamine infusion improves oxygen delivery (Do2) and oxygen uptake (VO2) in hyperdynamic septic shock patients stabilized by adequate volume and dobutamine alone, or by the combination of dobutamine and norepinephrine. DESIGN: Prospective clinical trial of two patient groups. Group 1 (n = 15) was stabilized with dobutamine, and group 2 (n = 10) was stabilized with dobutamine and norepinephrine. SETTING: Intensive care unit in a university hospital. PATIENTS: Twenty-five postoperative, hyperdynamic septic shock patients. INTERVENTIONS: The stabilizing catecholamine infusion was replaced in a stepwise manner by dopamine to achieve a similar mean arterial pressure (dopamine doses: group 1, mean 22 +/- 15 micrograms/kg/min [range 6 to 52]; and group 2, mean 57 +/- 41 micrograms/kg/min [range 15 to 130]). MEASUREMENTS AND MAIN RESULTS: A complete hemodynamic profile was performed with oxygen transport related variables at baseline, after replacement by dopamine, and after resetting to the original catecholamine infusion. The change to dopamine resulted in increases in cardiac index (group 1: 20% [p < .01]; group 2: 33% [p < .01]), and DO2 (group 1: 19% [p < .01]; group 2: 27% [p < .01]). However, VO2, whether directly measured from the respiratory gases or calculated by the cardiovascular Fick principle, did not change in both groups with dopamine, while the oxygen extraction ratio decreased significantly in both groups with dopamine. Heart rate, pulmonary artery occlusion pressure, and pulmonary shunt fraction all increased with dopamine. PaO2 decreased, but oxygen saturation remained stable in both groups with dopamine. CONCLUSIONS: Short-term dopamine infusion in hyperdynamic septic shock patients, despite producing higher global DO2, was not superior to dobutamine or the combination of dobutamine and norepinephrine infusion. PMID- 7497717 TI - Catecholamine and cortisol responses to lower extremity revascularization: correlation with outcome variables. Perioperative Ischemia Randomized Anesthesia Trial Study Group. AB - OBJECTIVE: To determine whether catecholamine and cortisol secretory responses to surgery contribute to postoperative complications. DESIGN: Prospective, randomized, case series. SETTING: A university hospital operating suite and surgical intensive care unit. PATIENTS: Sixty patients undergoing lower extremity vascular surgery. INTERVENTIONS: Patients were randomized to receive either epidural anesthesia/epidural opiate analgesia (regional anesthesia) or general anesthesia/intravenous patient-controlled analgesia (general anesthesia). MEASUREMENTS AND MAIN RESULTS: Anesthesia was managed according to a prospectively designed protocol. Hemodynamic parameters and plasma catecholamine concentrations were determined at specific intraoperative and postoperative time points. Intraoperative and postoperative urine samples were collected and analyzed for free cortisol concentrations. Outcomes evaluated were cardiac (nonfatal myocardial infarction and cardiac death) and surgical (graft occlusion). Mean arterial pressure during emergence from anesthesia and in the early postoperative period correlated positively with plasma norepinephrine concentration (p < .01). In addition, plasma catecholamine concentrations were higher in patients with postoperative hypertension. Plasma norepinephrine concentrations at the time of emergence from anesthesia and postoperatively were also higher in patients requiring repeat surgery for graft revision, thrombectomy, or amputation (p < .05). Multivariate analysis indicated that the norepinephrine concentration at the time of emergence, but not type of anesthesia, correlated with reoperation for graft occlusion, suggesting that the previously reported beneficial effect of regional anesthesia may be due to modulation of the stress response. Myocardial infarction or cardiac death occurred in three patients. These patients had markedly increased catecholamine concentrations. CONCLUSIONS: The catecholamine response to lower extremity vascular surgery contributes to the development of postoperative hypertension and may also be important in the development of thrombotic complications. PMID- 7497720 TI - Immunodepression following neurosurgical procedures. AB - OBJECTIVE: To determine the influence of a selective, sterile central nervous system surgery on immune reactivity, particularly whether a decrease of monocytic human leukocyte antigen-DR expression, indicating immunodepression, occurs after neurosurgery and if this measurement is useful for identification of patients with a high risk of infection. DESIGN: Prospective study. SETTING: Department of neurosurgery and intensive care unit in a university hospital. PATIENTS AND INTERVENTIONS: Blood samples were obtained from 46 patients at least once during the first 3 days after undergoing sterile central nervous system surgery. Fourteen of these patients developed infectious complications as defined by clinical and microbiological criteria. In ten of 46 patients, paired samples of blood and cerebrospinal fluid were collected from a ventricle drain at the following times: 1 day before surgery; several times on the day of surgery; and every day after surgery for at least 6 days. MEASUREMENTS AND MAIN RESULTS: Monocytic human leukocyte antigen-DR expression, as measured by flow cytometry on days 1 through 3 after surgery in 46 patients, was lower in 14 patients who developed infection after neurosurgery (p < .0001). In all ten closely monitored patients, monocytic human leukocyte antigen-DR expression decreased temporarily after surgery. Of these patients, only one patient showed a persistent and considerably decreased monocytic human leukocyte antigen-DR expression. This patient was the only patient in this subgroup who developed sepsis syndrome. In order to assess whether the monocytic human leukocyte antigen-DR decrease was associated with a preceding inflammatory response, local and systemic concentrations of interleukin (IL)-1 beta, IL-6, IL-8, tumor necrosis factor alpha, and interferon-gamma were measured in this subgroup. These cytokines were not detectable in plasma during the first days after surgery. In contrast, considerable increases of IL-6 and IL-8 concentrations were detectable in cerebrospinal fluid within hours after surgery. CONCLUSIONS: A decrease of monocytic human leukocyte antigen-DR expression occurs after neurosurgery and is associated with a preceding, strong, intracranial (but not systemic) inflammatory response. A very low monocytic human leukocyte antigen-DR expression (< 30% positive monocytes) suggests the possibility of infection. Measurement of monocytic human leukocyte antigen-DR expression could help to detect patients with a high risk of infection after neurosurgery. Our results suggest that even sterile central nervous system surgery may contribute to general immunodepression. The local intracranial inflammatory response may be involved in this process. PMID- 7497722 TI - Goal-directed therapy with dopexamine, dobutamine, and volume expansion: effects of systemic oxygen transport on hepatic ultrastructure in porcine sepsis. AB - OBJECTIVES: Can the hepatic structural deterioration that occurs during peritonitis be attenuated by increasing cardiac output and oxygen consumption (VO2)? Do the agents used to achieve these increases have any characteristic affects on these hepatic structural changes? DESIGN: Randomized, prospective, observational animal study. SETTING: Research laboratory of a university medical school. SUBJECTS: Twenty-five Middle White adolescent pigs, weighing 25 to 30 kg, divided into five groups. INTERVENTIONS: A thermodilution flotation catheter was advanced into the pulmonary artery. Additional catheters were inserted into the jugular, portal, and hepatic veins, and into the femoral artery. Ultrasound flow probes were placed around the portal vein and the hepatic artery. A metabolic cart was attached to the ventilator. Baseline measurements were made and cardiac output was increased by > 25% by administering either dobutamine (10 micrograms/min), dopexamine (10 micrograms/kg/min), or colloid. A control group had its cardiac output maintained at its baseline value. Peritonitis was induced in the four groups by contamination with cecal content and maintained for 6 hrs. Hepatic tissue was then removed for ultrastructural analysis and the animals were killed. MEASUREMENTS AND MAIN RESULTS: Before infection, cardiac output, VO2, and hepatic blood flow were increased in the three treatment groups. In the dobutamine and dopexamine groups, oxygen delivery increased, but decreased in the volume group. Mean arterial pressure increased in the dobutamine and dopexamine groups, but in the volume group, mean arterial pressure was maintained. Six hours after infection, cardiac output and VO2 had further increased in the dobutamine and volume groups, but both variables had decreased in the dopexamine group. After infection in the control group, cardiac output had decreased, although oxygen delivery and VO2 increased. There were no significant differences between hepatic hemodynamic or oxygen transport variables in any of the groups during the infection period. Hepatic ultrastructure was well maintained in the dopexamine group, while considerable deterioration was seen in the volume and control groups. In the dobutamine group, hepatic deterioration was greater than in the other three groups. CONCLUSIONS: Increasing cardiac output and VO2 before and during infection was only protective when dopexamine was administered. Dobutamine infusion was associated with greater hepatic deterioration than that effect seen in either the control or volume groups. PMID- 7497721 TI - Systemic perfusion pressure and blood flow before and after administration of epinephrine during experimental cardiopulmonary resuscitation. AB - OBJECTIVES: To evaluate instantaneous blood flow variations in the compression and relaxation phases of cardiopulmonary resuscitation (CPR) and the effect of epinephrine administration. DESIGN: Prospective, randomized, controlled trial. SETTING: Experimental laboratory in a university hospital. SUBJECTS: Twenty-two anesthetized piglets. INTERVENTIONS: A tracheostomy was performed and arterial, central venous, and pulmonary arterial catheters were inserted, followed by thoracotomy with placement of pulmonary arterial, aortic, and left anterior descending coronary arterial (extended study group) flow probes and a left atrial catheter. Ventricular fibrillation for 2 mins was followed by 10 mins of either open-chest (n = 10) or closed-chest (n = 12) CPR. Seven minutes after the initiation of CPR, all piglets received 0.5 mg of epinephrine iv; at 12 mins, direct current shocks were applied. MEASUREMENTS AND MAIN RESULTS: Open-chest CPR generated greater systemic perfusion pressure than closed-chest CPR, especially during the relaxation phase, resulting in greater mean blood flow. With both open and closed-chest CPR, antegrade pulmonary arterial and aortic blood flow occurred during compression, whereas antegrade left anterior descending coronary arterial blood flow occurred during relaxation. During relaxation, retrograde flow was found in the pulmonary artery and aorta. During compression, retrograde flow was found in the left anterior descending coronary artery. The administration of epinephrine had the following effects: a) increased the systemic perfusion pressure more during open- than closed-chest CPR; b) increased the systemic relaxation perfusion pressure more than the compression perfusion pressure; c) decreased mean pulmonary arterial and aortic blood flow, but substantially increased the mean left anterior descending coronary artery blood flow; and d) reduced the retrograde flow in the left anterior descending coronary artery. CONCLUSIONS: Open-chest CPR generated greater systemic perfusion pressure and blood flow than closed-chest CPR. Epinephrine increased left anterior descending coronary artery blood flow but decreased total cardiac output, such that cerebral perfusion might be endangered. This problem will be studied separately. PMID- 7497723 TI - Ketamine attenuates endotoxin-induced leukocyte adherence in rat mesenteric venules. AB - OBJECTIVES: To determine the influence of ketamine on endotoxin-induced leukocyte adherence and venular microhemodynamics. DESIGN: Randomized, controlled trial. SETTING: Experimental laboratory. SUBJECTS: Thirty male Wistar rats. INTERVENTIONS: The rats were pretreated with ketamine (10 mg/kg iv) or 0.9% saline, and both groups were given endotoxin (Escherichia coli lipopolysaccharide; 5 mg/kg iv). The control group received two doses of 0.9% saline. MEASUREMENTS AND MAIN RESULTS: The rates of leukocyte adherence and changes in microhemodynamics were monitored in rat mesenteric venules, using in vivo video microscopy. The number of adherent leukocytes was determined on-line in 10-min intervals from 60 mins before until 2 hrs after endotoxin administration. Venular diameters, red blood cell velocity, volumetric blood flow, and the venular wall shear rate were monitored before and at 10, 30, and 60 mins after endotoxin exposure. A 6.3-fold increase in the number of adherent leukocytes was observed 10 mins after administration of endotoxin when compared with control animals (5.87 +/- 0.69 vs. 0.93 +/- 0.21 adherent cells/100 microns; p < .001). This increase remained unchanged for 120 mins. In ketamine-pretreated rats, a 2.6-fold increase in leukocyte adherence occurred during the first 20 mins after endotoxin exposure (2.40 +/- 0.46 vs. 0.93 +/- 0.21 adherent cells/100 microns; p < .01). However, no difference in the number of adherent leukocytes between ketamine-pretreated and control animals was found after this 20-min period. In animals of the control group, no increase in leukocyte adherence occurred during the entire observation time. Diameters of mesenteric venules did not change after endotoxin exposure in any of the groups. Red blood cell velocity and venular blood flow in the endotoxin-treated groups decreased 10 mins after the injection of endotoxin when compared with controls, but these values did not show any difference when they were compared between ketamine and saline pretreated animals. Similarly, venular wall shear rate in the endotoxin-treated groups decreased 10 and 30 mins after injection of endotoxin. However, no significant difference occurred between ketamine and saline-pretreated animals. CONCLUSIONS: Pretreatment with ketamine attenuates endotoxin-induced leukocyte adherence by a shear rate-independent mechanism, suggesting reduced expression of adhesion molecules. These results indicate that ketamine exerts an anti inflammatory effect, which might be beneficial in septic patients. PMID- 7497725 TI - Extravasation rates and complications of intraosseous needles during gravity and pressure infusion. AB - OBJECTIVE: To compare the extravasation rates and insertion complications under gravity and 300 mm Hg (40 kPa) pressure infusion of threaded (SurFast and Sussmane-Raszynski intraosseous needles, Cook Critical Care, Bloomington, IN); and nonthreaded needles (16-gauge disposable intraosseous needle with 45 degrees trocar Cook Critical Care, Bloomington, IN; Jamshidi bone marrow needle; Baxter Health Care Corp, Valencia, CA). DESIGN: A prospective, randomized study. SETTING: An animal laboratory at a university center. SUBJECTS: Five healthy mix breed piglets, weighing 15 to 15.5 kg. INTERVENTIONS: Piglets were anesthetized and ventilated. Tibial, femoral, and humeral osseous sites were exposed by dissection of overlying tissue. All bleeding points were cauterized and oozing was prevented by sealing with cyanoacrylate. Intraosseous access devices then were inserted one at a time in random order and rated for difficulty of insertion. Normal saline solution was infused under gravity or 300 mm Hg (40 kPa) pressure. Extravasation rates then were calculated from the increase in weight of a gauze sponge wrapped tightly at the base of the needle during infusion. MEASUREMENTS AND MAIN RESULTS: No significant (p > .05) differences in extravasation rates were noted among the different types of needles, either under gravity or pressure infusions. The Sussmane-Raszynski needle was significantly more difficult to insert than the others (rated difficult to insert and control in 16 of 34 attempts). Inadvertent penetration of both cortices occurred with nonthreaded needles only (three of 66 attempts). The SurFast needle provided greatest penetration control and was most resistant to accidental dislodgement. CONCLUSIONS: Under ideal conditions, needle type does not influence extravasation rates. However, difficulty with insertion and penetration of both cortices occur commonly and may lead to extravasation during stressful emergency situations or when performed by unskilled personnel. PMID- 7497724 TI - Comparison of transtracheal and extravascular Doppler determinations of stroke volume and cardiac output at various states of volume loading in piglets. AB - OBJECTIVE: To assess the applicability of a new technology in neonates. Transtracheal Doppler and extravascular Doppler determinations of stroke volume and cardiac output were compared with thermodilution measurements at various states of volume loading in an animal model. DESIGN: Prospective, descriptive study. SETTING: Animal research laboratory at a university medical center. SUBJECTS: Fourteen newly weaned piglets, weighing 2.8 to 6.5 kg. INTERVENTIONS: Doppler probes were placed on the endotracheal tube tip (transtracheal Doppler) and directly on the aortic adventitia (extravascular Doppler). A 4-Fr thermodilution catheter was inserted in the pulmonary artery. Stroke volume and cardiac output determinations were recorded at baseline, after a 15-mL/kg volume load and after successive 15-mL/kg blood withdrawals to exsanguination or a systolic blood pressure of < 20 mm Hg. MEASUREMENTS AND MAIN RESULTS: Transtracheal and extravascular Doppler measurements of cardiac output were not significantly different from thermodilution at any physiologic state. These techniques were able to measure stroke volumes and cardiac outputs at the low levels seen in severe hemorrhagic shock. CONCLUSIONS: Transtracheal Doppler and extravascular Doppler measurements of cardiac output compare favorably with thermodilution. These methods effectively followed trends from alterations in intravascular volume, even at very high heart rates and small stroke volumes. Transtracheal Doppler and extravascular Doppler should yield useful information in critically ill neonatal patients, where data regarding stroke volume and cardiac output may be useful in clinical management. PMID- 7497727 TI - Sedation for the critically ill neurologic patient. AB - OBJECTIVE: To review the scientific basis for sedation of critically ill neurologic patients by summarizing the distinct neurophysiologic disturbances present in this population and presenting the central nervous system effects of sedative agents to permit optimal drug therapy. DATA SOURCES: Review of the English language clinical and scientific literature using MEDline data search. STUDY SELECTION: Literature references were selected through a key word search of sedative therapy, drugs used for sedation, and specific neurologic disorders and processes to provide an in-depth overview of sedative drug mechanisms of action, effects on neurophysiology and intracranial dynamics, pharmacokinetics, and toxicity profile. Special emphasis was placed on neurologic side effects. DATA EXTRACTION: Clinical and scientific literature was reviewed and data relevant to neurophysiologic effects of sedative drug therapy were summarized. Recommendations for institution of sedative therapy and of particular agents were made as a result of analysis of all pooled data. DATA SYNTHESIS: Critically ill patients with neurologic pathology present as a unique subset of individuals cared for in an acute care setting. Because monitoring of neurologic patients requires frequent assessment of the neurologic examination, the goal of sedative therapy should be to enhance, or to minimally perturb elicitation of the examination. Neurophysiologic disturbances introduce distinct risks for sedation and require their identification and understanding before the initiation of any sedative therapy. Sedative drugs, in particular, act to disturb central nervous system function and their effects may result in diagnostic confusion and further neurologic deterioration. The pharmacokinetic and neurophysiologic actions of the common classes of sedative agents, such as benzodiazepines, opioids, barbiturates, and neuroleptics, as well as ketamine, propofol, and clonidine are discussed. Recommendations are presented based on the specific type of sedation required and the underlying neurologic disturbance. Several specific examples, including head trauma, neuromuscular disease, and alcohol withdrawal, are provided. CONCLUSIONS: Preservation of the neurologic examination is paramount in documenting clinical improvement or deterioration in the critically ill neurologic patient. Pharmacologic sedation in this unique population of acute care patients requires careful consideration of the underlying neurophysiologic disturbances and potential adverse effects introduced by sedative drugs. PMID- 7497728 TI - Remote communication from a mobile terminal: an adjunct for a computerized intensive care unit order management system. AB - OBJECTIVES: To develop and implement a fully mobile computer terminal that interfaces with our computerized intensive care unit (ICU) local area network and order management system. This system can provide access to the entire network and to order review and entry and during ICU bedside rounds. DESIGN: Descriptive report. SETTING: Surgical ICU in Department of Veterans Affairs Medical Center. SYSTEM CONFIGURATION: A parallel local area network was configured for the remote mobile computer system. A proprietary remote transmission system (Altair II, Motorola) was used. This high-throughput system minimizes interference and errors by using licensed, nonshared, radiofrequency spectra. CONCLUSION: The resulting mobile system is an economical and time-efficient adjunct to an established ICU computerized network and order management system. Clinical working bedside rounds are now routinely conducted with the mobile terminal, providing immediate access to full network resources. PMID- 7497726 TI - A canine study of cold water drowning in fresh versus salt water. AB - OBJECTIVE: To compare the pathophysiologic changes occurring during drowning in cold fresh water and cold salt water with reference to viability. DESIGN: Randomized, prospective, controlled submersion experiments in two contrasting cold liquids. SETTING: A laboratory at a large university-affiliated medical institution. SUBJECTS: Thirteen healthy, anesthetized mongrel dogs. Three dogs served as controls and were immersed but not submerged. The remainder were submerged in cold fresh water or cold salt water (4 degrees C). INTERVENTIONS: Catheters were placed in the femoral artery, right carotid artery and right internal jugular vein. Electrocardiogram, pneumogram, and rectal temperatures were measured continuously during submersion/immersion. MEASUREMENTS AND MAIN RESULTS: Cold water submersion with drowning produced a large initial decrease in carotid artery temperature (approximately 7.5 degrees C in the first 2 mins) compared with a minor decrease (approximately 0.8 degrees C with immersion). No significant differences were noted in the rate of decrease of temperature between drowning in fresh water and salt water. During cold fresh water drowning, aspiration produced gross hemodilution with an average increase in body weight of 16.5%. Hematocrit values, serum sodium concentrations, and osmolality decreased while serum potassium concentrations, catecholamines, and free hemoglobin increased. All measured biochemical data (except PaO2) remained at viable levels. By contrast, during cold salt water drowning, average body weight increased by only 6%, with hemoconcentration and a shrinkage of vascular volume. Hematocrit and hemoglobin values increased by 30%, but initial plasma free hemoglobin values remained unchanged. Serum sodium concentrations, osmolality, and potassium concentrations increased rapidly to critical levels. CONCLUSIONS: On submersion in cold water, all of the experimental animals developed tachypnea immediately, followed by aspiration with predictable effects. The biochemical and pathophysiologic changes in cold water drowning approximated those changes reported for warm water drowning for both fresh and salt water with one exception and continued aspiration of cold water produced extremely rapid core cooling as long as the circulation remained intact. This process of acute submersion hypothermia may protect the brain temporarily from lethal damage, as reported in cases of cold fresh water drowning. Concentrations of circulating catecholamines increased exponentially in both groups of test animals. Clinically, their acute effects on the circulation, compounded by significant hypothermia and extreme anoxia, must hamper the detection of residual circulation at rescue and may play a role in sudden death from cold water in the absence of drowning. PMID- 7497729 TI - Use of T-test with nonparametric data: fentanyl withdrawal syndrome. PMID- 7497730 TI - Alienation and the production of strangers: Western medical epistemology and the architectonics of the body. An historical perspective. AB - A critical history of the architectonics of the Western body and the scientific methodologies which underlay its construction in late seventeenth England offers insight into current experiences of alienation and estrangement on the part of a substantial number of patients and health-care providers. The bodily architectonics outlined formed normative personae that continue to shape a number of medical and social discourses. This essay outlines some features of this bodily architecture and summarizes some of the factors that helped shape its historical formation. Discussion then moves to consideration of those who opposed the new formulations and/or methods, and finally suggests some ways in which this bodily architecture remains both current and problematic today. PMID- 7497731 TI - "True clinical fictions": medical and literary narratives from the Civil War Hospital. AB - Where to find stories of the "real" Civil War? Since Walt Whitman, observers have suggested the place to look is the military hospital. This paper offers a comparative analysis of medical case histories and literary stories written by S. Weir Mitchell, head contract-surgeon at one of the largest Union army research hospitals. I illustrate that Mitchell's case histories and stories were at once complementary and antagonistic in their use of medical knowledge and narrative technique as a means of achieving truth-value. PMID- 7497732 TI - On the vagaries of bodies. PMID- 7497733 TI - The science of the humane and the humanism of science. PMID- 7497734 TI - Violence, solace, and ritual: a case study from island Southeast Asia. AB - Most headhunting traditions in island Southeast Asia link ritual violence to grief and mourning. Some of the more persuasive analyses of these practices pivot on notions of rage and catharsis, arguing that turbulent emotions motivate persons to take up cleansing acts of violence. This paper seeks a more complex understanding of how ritual may connect bereavement and violence through a look at case materials from highland Sulawesi (Indonesia). Ritual practices there suggest that the resolution of communal mourning is more significant than personal catharsis in motivating violence; that individual affect is refigured collectively as "political affect;" and that varied discursive forms, such as vows, songs, and noise mediate the ways in which people put grief behind them and resume their lives. Indeed, such discursive forms appear to be generative sites for violence and solace. PMID- 7497735 TI - Alcoholism in Ghana--a socio-cultural exploration. AB - A vast literature has accumulated in recent years, examining the disease concept of alcoholism, and analyzing the interaction of biomedicine with indigenous healing systems in colonial and post-colonial societies. Social scientists have consistently emphasized the social context of alcoholism, although their works have been largely ignored. This article engages the literature on the social history of medicine in Africa, and works on alcohol use in non-Western societies, in an attempt to offer an understanding of alcoholism in Ghana rooted in Ghanaian cultures and history. It explores how alcohol's established ties with spirituality influences Ghanaian perceptions of alcoholism. Based on interviews, highlife music, popular literature, and the few written works on alcohol use in Ghana, the article examines the social construction of the alcoholic in independent Ghana. PMID- 7497737 TI - Towards the development of a fluorescent in situ hybridization based genotoxicologic assay for aneuploidy detection in sperm. AB - Fluorescent in situ hybridization (FISH) is a molecular cytogenetic technique which allows an analysis of chromosome copy number in interphase cells. The cytological examination of spermatozoa is routinely performed as part of semen analysis. While this analysis is only a component of the evaluation of an infertile male, the aetiology of many cases of infertility remains unknown. FISH on semen smears can yield information on chromosome-specific aneuploidy due to constitutional and acquired chromosomal abnormalities as found with cancer or exposure to genotoxic substances. The ability to analyse germ cells directly is a useful adjunct to GTG-banding analysis of meiotic chromosomes, an invasive and extremely labour-intensive procedure. The work of 2 years in developing and optimizing a FISH-based assay in routinely processed semen smears is described. PMID- 7497736 TI - Sepsis in the critically ill patient. PMID- 7497738 TI - Detection of triploidy in fish using the cytokinesis-blocked method for erythrocyte and hepatic cells. AB - The use of two cytochalasin-blocked methods with erythrocyte and hepatic cells was developed to identify spontaneous and/or induced triploid fish by observing the presence of trinucleated cells. Spontaneous triploids were found in two specimens out of fifteen commercial rainbow trout from a hatchery at 2 years of age. Some rare spontaneous triploid (3n) trinucleated cells were also found in the other thirteen diploid (2n) trout specimens under investigation. The use of this novel technique in a study of the triploidy in vitro, allows evaluation of the polyploid specimens under field and laboratory experimental conditions. The potential contribution of binucleated hepatic and erythrocyte techniques in fish to the assessment of polyploidy is discussed. PMID- 7497739 TI - Effect of carbon dioxide on the temperature dependence of anion exchange in human red cells. AB - Anion exchange across the red cell membrane was studied by measuring the rate of shrinkage of cells when transferred from a medium of low pH to one of higher pH. Removal of trace amounts of CO2/bicarbonate from the media by degassing and the inhibition of carbonic anhydrase with 5 microM ethoxzolamide slowed the shrinkage rate. Arrhenius plots were linear over a temperature range of 40 degrees C, both in the presence of trace amounts of CO2/bicarbonate and in their absence, and gave an apparent activation energy for Cl- exchange of 73.9 +/- 8.9 kJ mole-1 when CO2 was present but this increased to 135.8 +/- 7.7 kJ mole-1 in its absence. From the results it is concluded that the lower value for the activation energy is determined by the rate of formation of bicarbonate ions while the high value represents hydroxyl:anion exchange and is a truer measure of the activation energy of the exchange process. PMID- 7497740 TI - Do we know the risks? PMID- 7497741 TI - Management of acute inter-atrial shunting. PMID- 7497742 TI - Acute myocardial infarction. Shifting paradigms of diagnosis and care in a cost conscious environment. PMID- 7497743 TI - Inhaled corticosteroids in COPD. A light at the end of the tunnel? PMID- 7497744 TI - Technology assessment. PMID- 7497745 TI - Quality of life after lung transplantation. PMID- 7497747 TI - The gastrointestinal tract and critical illness. Is more perfusion better? PMID- 7497748 TI - Invasive diagnostic methods for nosocomial pneumonia. Revisiting the folklore. PMID- 7497749 TI - Plaque abrasion and intra-aortic balloon leak. AB - STUDY OBJECTIVE: To determine and correct cause of high incidence of intra-aortic balloon leaks (ruptures). DESIGN: Epidemiologic investigation of factors associated with intra-aortic balloon leak, and sequential application of corrective measures evaluated by continued concurrent data collection. SETTING: Thirty-four-bed ICU in 598-bed tertiary care medical center. INTERVENTIONS: Procedure changed to place smaller balloons (34 mL instead of 40 mL) in patients less than 163 cm in height. MEASUREMENTS AND RESULTS: Demographic and clinical data on all patients showed no change after initial interventions, followed by significant drop (8 to 2%) in incidence of balloon leak when smaller, shorter balloons were placed in shorter patients. CONCLUSIONS: Placement of larger, longer balloons in patients increases risk of perforation of balloon by calcific plaque in the distal thoracic and abdominal aorta. PMID- 7497746 TI - Is mouth-to-mouth ventilation necessary for successful resuscitation? PMID- 7497750 TI - Percutaneous balloon pericardiotomy for malignant pericardial tamponade. AB - Recurrent pericardial effusion or tamponade can complicate malignant disease. The medical approach with pharmacologic agents has a high recurrence rate. Balloon pericardiotomy (BP) may provide a less invasive alternative to surgery. We performed BP in ten high-risk patients with malignant pericardial tamponade (MPT). BP was performed with aseptic technique under fluoroscopic and echocardiographic guidance. We used a percutaneous subxiphoid approach. A 20- to 25-mm-wide and 4-cm-long balloon was introduced over a stiff guide wire, positioned across the parietal pericardium, and manually inflated until the waisting disappeared. Echocardiographic study and chest radiograph were taken before, immediately after the procedure, and during follow-up. The procedure was successful and well tolerated in all patients. There were no immediate or late complications. No patients developed recurrence of pericardial effusion at up to 10 months' follow-up. In conclusion, BP can be performed in patients with MPT with high rate of procedural success. It may become the preferred treatment to avoid a more invasive procedure in these critically ill patients. PMID- 7497754 TI - Hemorrhagic complications of thrombolytic therapy. AB - Thrombolytic therapy salvages ischemic myocardium by rapidly reestablishing coronary artery patency in acute myocardial infarction. One of its major limitations is the complication of hemorrhage. A retrospective study of myocardial infarction patients who received thrombolytic therapy was performed to determine risk factors associated with a hemorrhagic event. Three hundred fifty patients were enrolled, and 20 (5.7%) had a bleeding complication, including four patients (1.1%) who had an intracranial hemorrhage. The factors associated with an increased risk for a significant hemorrhagic event were age (> 65 years) and female gender. Factors associated with an intracranial hemorrhage were age (> 65 years) and a history of hypertension. PMID- 7497751 TI - A decision tree for the early diagnosis of acute myocardial infarction in nontraumatic chest pain patients at hospital admission. AB - STUDY OBJECTIVE: To find an accurate algorithm for the diagnosis of acute myocardial infarction in nontraumatic chest pain patients on presentation to the emergency department. DESIGN: In a prospective clinical study, we compared the diagnostic performances of clinical symptoms, presenting ECG, creatinine kinase, creatine kinase MB activity and mass concentration, myoglobin, and cardiac troponin T test results of hospital admission blood samples. By classification and regression trees, a decision tree for the diagnosis of acute myocardial infarction was developed. SETTING: Emergency room of a Department of Internal Medicine (University Hospital). PATIENTS: One hundred fourteen nontraumatic chest pain patients (median delay from onset of chest pain to hospital admission, 3 h; range, 0.33 to 22): 26 Q-wave and 19 non-Q-wave myocardial infarctions, 49 patients with unstable angina pectoris, and 20 patients with chest pain caused by other diseases. MEASUREMENTS AND RESULTS: Of each parameter taken by itself, the ECG was tendentiously most informative (areas under receiver operating characteristic plots: 0.87 +/- 0.04 [ECG], 0.80 +/- 0.08 [myoglobin], 0.80 +/- 0.04 [creatine kinase MB mass], 0.77 +/- 0.04 [creatine kinase activity], 0.69 +/ 0.06 [clinical symptoms] 0.67 +/- 0.06 [creatine kinase MB activity], 0.67 +/- 0.05 [troponin T]). In patients presenting 3 h or less after the onset of chest pain, ECG signs of acute transmural myocardial ischemia were the best discriminator between patients with and without myocardial infarction. In patients presenting more than 3 h, however, creatine kinase MB mass concentrations (discriminator value, 6.7 micrograms/L) were superior to the ECG, clinical symptoms, and all other biochemical markers tested. This algorithm for diagnosing acute myocardial infarction was superior to each parameter by itself and was characterized by 0.91 sensitivity, a 0.90 specificity, a 0.90 positive and negative predictive value, and a 0.90 efficiency. CONCLUSIONS: We found an algorithm that could accurately separate the myocardial infarction patients from the others on admission to the emergency department. Therefore, this classifier could be a valuable diagnostic aid for rapid confirmation of a suspected myocardial infarction. PMID- 7497753 TI - Lack of association of higher insulin levels with diffuse atherosclerotic coronary artery disease in nondiabetics. AB - Since there is experimental evidence that insulin promotes atherosclerosis, we tested the hypothesis that insulin levels are higher in patients with diffuse atherosclerotic coronary artery disease by measuring insulin levels in 46 nondiabetic patients with angiographically defined diffuse coronary artery disease and 46 normal controls with angiographically normal coronary arteries. Fasting insulin levels were similar in both groups of patients: 7.70 +/- 5.77 microU/mL in those with diffuse coronary disease versus 7.39 +/- 5.01 microU/mL in controls. Also, insulin levels drawn 1 and 2 h after oral glucose challenge were not significantly different in patients with diffuse disease (48.78 +/- 32.46 microU/mL and 42.26 +/- 32.38 microU/mL, respectively) compared with patients with normal coronary arteries (51.03 +/- 28.01 microU/mL and 43.79 +/- 31.62 microU/mL, respectively). We conclude that insulin probably does not promote clinical atherosclerosis in nondiabetics. PMID- 7497752 TI - Gender differences in clinical evaluation and triage in coronary artery disease. AB - BACKGROUND: Women undergo evaluation and treatment for cardiac diseases less frequently than men with similar symptoms. The purpose of this study was to determine what differences exist in clinical evaluation and treatment between men and women presenting with coronary heart disease that may indicate a gender bias. METHODS: A single hospital retrospective review of patients admitted with the diagnosis of cardiac ischemic syndrome, undergoing stress testing, coronary arteriography, percutaneous interventional procedure, or coronary bypass surgery was performed, including an evaluation by gender of the demographic, clinical, and angiographic parameters of 1 year of patients undergoing hospital admission, evaluation, or revascularization therapy for coronary heart disease in a single university hospital. RESULTS: Women admitted to the coronary care unit with a coronary diagnosis were less likely to undergo coronary arteriography than men. Women having a positive stress test result were as likely to undergo coronary arteriography as men with similar findings. Women undergoing coronary arteriography were as likely as men to undergo percutaneous transluminal coronary angioplasty, but less likely to undergo coronary artery bypass surgery. CONCLUSIONS: A gender-based selection bias exists in choosing patients to undergo coronary arteriography and coronary artery bypass grafting. PMID- 7497755 TI - Pharmacologic and hemodynamic effects of combined beta-agonist stimulation and phosphodiesterase inhibition in the failing human heart. AB - STUDY OBJECTIVES: We measured the individual and combined effects of the beta agonist dobutamine and the phosphodiesterase inhibitor enoximone both in vitro and in vivo in the failing human heart. DESIGN: This was an unblinded, prospective study. SETTING AND PATIENTS: The in vitro measurements were performed on 20 hearts obtained from subjects with end-stage biventricular failure and from seven normal hearts. The in vivo measurements were performed in eight subjects with class IV heart failure. INTERVENTIONS AND MEASUREMENTS: The in vitro measurements of enoximone, dobutamine, and the combination of these agents were phosphodiesterase activity using a sarcoplasmic reticulum-enriched preparation, cyclic adenosine monophosphate (cAMP) accumulation using particulate fractions, and tension response using isolated right ventricular trabeculae. The dose response to dobutamine, the combination of enoximone and dobutamine, and the combination of nitroprusside and dobutamine were measured in vivo using invasive hemodynamic monitoring. RESULTS: In vitro, enoximone exhibited dose-dependent inhibition of phosphodiesterase activity. The addition of enoximone to dobutamine resulted in an upward and leftward shift of the dobutamine dose-response curve for both cAMP production and contractile response. In vivo, enoximone significantly shifted the dobutamine dose-response curves for cardiac index, left ventricular stroke work index, and heart rate upward and to the left; and shifted the dobutamine dose-response curves for right atrial, pulmonary arterial, and pulmonary wedge pressures downward and to the right. CONCLUSIONS: Enoximone exerts favorable effects on cardiac performance that are additive to those produced by dobutamine. These effects are mediated by increasing cellular cAMP concentrations through independent, additive mechanisms. PMID- 7497756 TI - Abnormal ventricular long-axis function in systemic sclerosis. AB - STUDY OBJECTIVE: To assess possible effects of systemic sclerosis on ventricular function. DESIGN: Retrospective analysis of patients referred for echocardiographic examination to assess ventricular function. SETTING: Tertiary referral center for cardiac and chest diseases equipped with invasive and noninvasive facilities. PATIENTS: Thirty-four patients with clinical diagnosis of systemic sclerosis, aged 49 +/- 12 years; 24 had pulmonary fibrosis and 10 did not. There were 21 normal controls of similar age. MEASUREMENTS: Two-dimensional guided M-mode echocardiographic recordings of the left ventricular minor and long axis at the left and septal sites and right ventricle were obtained. Transmitral and transtricuspid Doppler flow velocities were also obtained with ECG and phonocardiogram. RESULTS: In 24 patients with pulmonary fibrosis, long-axis excursion was reduced 2.1 +/- 0.5 vs 2.7 +/- 0.4 cm/s as was peak rate of shortening and lengthening, 8.5 +/- 3.3 vs 10.8 +/- 2.4 cm/s and 7.5 +/- 2.5 vs 12 +/- 3.6 cm/s, respectively (p < 0.001), at the right side compared with 10 patients without. The onset of right long-axis shortening and lengthening was delayed with respect to the Q wave of the ECG and P2 of the phonocardiogram (p < 0.001 in both vs controls). The onset of tricuspid forward flow from the second heart sound was also delayed in the two groups, 110 +/- 15 ms and 100 +/- 20 ms vs 80 +/- 15 ms, respectively (p < 0.001). Right ventricular late diastolic filling velocities were increased 35 +/- 15 and 35 +/- 12 cm/s vs 20 +/- 10 cm/s in both groups (p < 0.01), and hence E:A ratio reduced 1.25 +/- 0.5 and 1.4 +/- 0.3 vs 1.9 +/- 0.4, respectively (p < 0.001). Pulmonary flow acceleration time was reduced only in patients with pulmonary fibrosis, 105 +/- 30 ms vs 125 +/- 30 ms (p < 0.001). At the left side, total long-axis excursion was reduced only in patients with pulmonary fibrosis (p < 0.01), while peak shortening and lengthening rates were reduced in both groups (p < 0.05). The onset of shortening from the Q wave and lengthening from the second heart sound were both delayed in the two groups with the latter greatly delayed in patients with pulmonary fibrosis (p < 0.05). CONCLUSIONS: Right and left ventricular long-axis function is frequently abnormal in patients with systemic sclerosis. Abnormalities are more profound in patients with CT evidence of pulmonary fibrosis than in those without. We suggest that these disturbances are due to myocardial fibrosis which, from the anatomic distribution of longitudinally directed fibers, is likely to have been subendocardial. PMID- 7497757 TI - Cardiopulmonary bypass temperature does not affect postoperative euthyroid sick syndrome? AB - STUDY OBJECTIVE: To determine if temperature during cardiopulmonary bypass (CPB) has an effect on perioperative and postoperative thyroid function. DESIGN: Prospective study comparing thyroid function during and after hypothermic and normothermic CPB. SETTING: Cardiac surgical unit at a university-affiliated hospital. PATIENTS: Twelve patients scheduled to undergo cardiac operations with normothermic (n = 6) or hypothermic (n = 6) CPB. INTERVENTIONS: Blood was analyzed for serum concentration of total thyroxine (TT4), total triiodothyronine (TT3), free T3 (fT3), reverse T3 (rT3), and thyroid stimulating hormone (TSH) preoperatively, 60 min after CPB was initiated, 30 min after discontinuing CPB, and on postoperative days (POD) 1, 3, and 5. MEASUREMENTS AND RESULTS: Patients who underwent either cold (26 degrees +/- 5 degrees C) or warm (35 degrees +/- 1 degree C) CPB were comparable with regard to age, body weight, duration of CPB, cross-clamp time, use of inotropes, total heparin dose, and length of hospital stay. Incidence of postoperative myocardial infarction, congestive heart failure, and death were similar. In both groups, TT4 and TT3 were reduced below baseline values beginning with CPB and persisting for up to 5 days after CPB (p < 0.05), free T3 was reduced for up to 3 days after CPB (p < 0.05), mean serum rT3 was elevated on POD 1 and POD 3 (p < 0.05), and TSH remained unchanged. CONCLUSION: The results of this study suggest that normothermic CPB does not prevent the development of the "euthyroid sick syndrome" during and after CPB. Despite these changes in thyroid function, most patients in both groups had a normal postoperative recovery. PMID- 7497758 TI - Long-term results of coronary artery bypass surgery in patients with severely depressed left ventricular function. AB - STUDY OBJECTIVE: The objective of the present study was to evaluate medium- and long-term results of coronary artery bypass grafting (CABG) in patients with severe left ventricular dysfunction (LVD). DESIGN: Prospective evaluation (clinical follow-up and equilibrium radionuclide angiography scan) of all the patients with severe LVD who underwent CABG from November 1986 to November 1991 at the Tel Aviv Medical Center and were referred to the Post Cardiac Surgery Follow-up Clinic at this institution. PATIENTS: Seventy-four consecutive patients (65 men, 9 women, aged 43 to 82 years; mean age, 68.2 years) with left ventricular ejection fraction (LVEF) of 30% or less who underwent isolated CABG (without automatic implantable cardioverter-defibrillator implantation, aneurysmectomy, valve replacement, or other open heart procedures) during a 5 year period and were discharged from hospitalization were prospectively evaluated. Preoperatively, 62% of patients had angina, 65% had congestive heart failure (CHF), and the LVEF ranged from 10 to 30%. The mean number of grafts was 2.98 per patient; the internal mammary artery (IMA) was used in 54 patients. The patients were followed up 4 to 96 months (mean, 64.9 months) post-surgery for survival, clinical status, and left ventricular function. RESULTS: Survival was 96%, 93.2%, 91.9%, 87.8%, 86.5%, 83.8%, and 83.8%, at 1, 2, 3, 4, 5, 6, and 7 years, respectively. Postoperatively, mean angina class improved from 2.9 to 1.4 (p < 0.0001) and mean CHF class improved from 2.7 to 1.8 (p < 0.0001). Mean LVEF improved from 23.5% preoperatively to 35.7% postoperatively (p < 0.0001). CONCLUSIONS: The following occur in patients with coronary artery disease and severe LVD undergoing CABG: (1) good medium- and long-term survival is attained; (2) angina class improves; (3) CHF class improves; (4) LVEF objectively improves; and (5). IMA can be used safely as a conduit. PMID- 7497759 TI - Acute lung injury during cardiopulmonary bypass. Are the neutrophils responsible? AB - To test the hypothesis that acute lung injury during cardiopulmonary bypass (CPB) is related to the activation of neutrophils and the body temperature during bypass, we determined the differential WBC count, plasma elastase concentrations, and lung function before, during, and after CPB in 38 patients undergoing elective coronary artery bypass surgery. The patients were randomly assigned to receive either normothermic (n = 19, rectal temperature: 35.9 +/- 0.1 degrees C, mean +/- SE) or hypothermic (n = 19, 29.2 +/- 0.5 degrees C) CPB. The cellular response to the extracorporeal circulation was significantly delayed in the hypothermic group with a later onset of neutrophilia and a later increase in plasma elastase levels during bypass. Lung function deteriorated significantly after CPB as assessed by respiratory index, alveolar-arterial oxygen gradient, and intrapulmonary shunt, independent of bypass temperature. There was a positive correlation between peak elastase concentrations and postoperative respiratory index as well as intrapulmonary shunt (R2 = 0.5, p = 0.002 and R2 = 0.45, p = 0.003, respectively). Besides peak plasma elastase levels, multiple regression revealed no significant influence of other independent factors on postoperative lung dysfunction in our patients. PMID- 7497760 TI - Cigarette smoking and incidence of chronic bronchitis and asthma in women. AB - STUDY OBJECTIVE: To examine the relation of smoking habits and development of asthma in a large cohort of US women. DESIGN: Prospective cohort study. PARTICIPANTS: Among 74,072 women, 34 to 68 years of age, who were free of major diseases, we documented 671 incident asthma cases and 798 incident cases of chronic bronchitis during 10 years of follow-up. METHODS: Age-adjusted relative risk estimates for smoking categories were calculated separately for chronic bronchitis and asthma. RESULTS: Risk of chronic bronchitis was significantly higher in current smokers than in never smokers (relative risk [RR] = 2.85; 95% confidence interval [CI] = 2.45 to 3.32) and increased with the number of cigarettes smoked per day (p for trend < 0.0001). Approximately 5 years after quitting, chronic bronchitis risk in past smokers approached that in never smokers. In contrast, current smokers were at significantly lower risk for asthma than women who never smoked (RR = 0.57; 95% CI = 0.46 to 0.71) and women who quit (RR = 0.50; 95% CI = 0.40 to 0.62), possibly because individuals with sensitive airways are less likely to become regular smokers, and smokers who develop respiratory symptoms of any etiology tend to quit. Asthma risk in past smokers initially increased compared with that in never smokers, possibly because of quitting prior to diagnosis in response to symptoms of any etiology, but decreased with time since quitting (p for trend = 0.007); within approximately 5 years, the risk did not differ between past and never smokers. CONCLUSION: These data suggest that smoking in adults may not be an independent cause of asthma but could exacerbate or be perceived as exacerbating asthma symptoms in susceptible individuals. PMID- 7497761 TI - The effects of pentoxifylline on oxygenation, diffusion of carbon monoxide, and exercise tolerance in patients with COPD. AB - Pentoxifylline has been reported previously in an unblinded study to improve oxygen saturation, treadmill walk time, and resting diffusion of carbon monoxide (Dco) in patients with COPD. We recruited 12 patients with moderate to severe COPD whose exercise capacity was limited by ventilation or who developed hypoxemia with exercise. Patients were randomized to receive pentoxifylline or placebo, each for a 12-week period in a prospective, double-blind, crossover design study, to assess the effects of pentoxifylline on oxygenation, resting Dco, and exercise tolerance using arterial blood gas analysis. Eleven patients with a mean FEV1 of 0.94 L and a mean Dco of 9.85 mL/min/mm Hg completed the study. One patient withdrew from the study after developing pneumonia. There were no significant differences in resting oxygenation, resting Dco, or spirometry after 12 weeks of pentoxifylline relative to placebo. The 12-min walk test and dyspnea index for activities of daily living were also not significantly different while taking pentoxifylline. Finally, at maximal exercise, there were no differences in workload attained, exercise duration, oxygen consumption, carbon dioxide production, minute ventilation, oxygen saturation, PO2, alveolar arterial oxygen pressure difference, or Borg score while taking pentoxifylline relative to placebo. We conclude that pentoxifylline does not improve oxygenation, resting Dco, exercise tolerance, or dyspnea in patients with moderate to severe COPD. PMID- 7497762 TI - Inhaled budesonide therapy for patients with stable COPD. AB - A significant minority of patients with COPD have favorable response to corticosteroid treatment. In addition, the benefit of corticosteroid treatment may be outweighed by the side effects. Long-term administration of inhaled steroids is a safe means of treatment. We hypothesized that treatment with high dose inhaled budesonide would improve clinical symptoms and pulmonary function in subjects with COPD, and that the response to inhaled beta 2-agonist will serve to individualize steroid responders. We compared a 6-week course of 800 micrograms/d inhaled budesonide with placebo, separated by 4 weeks when no medication was taken, in a double-blind crossover trial, in 8 patients responding to inhaled beta 2-agonist, and in 22 nonresponders with stable COPD. In six of eight "responders to beta 2-agonist," there was a significant improvement in the FEV1 (defined as > or = 20%) following inhaled budesonide, as compared with placebo. In the 22 "nonresponders to beta 2-agonist," there was no significant improvement in the mean FEV1 (1.41 +/- 0.1 L before, and 1.61 +/- 0.1 L after treatment) with inhaled budesonide or placebo. Over the 6-week course of treatment by either budesonide or placebo, the nonresponders reported similar beta 2-agonist consumption (4.8 +/- 0.2 and 5.0 +/- 0.1 puffs per patient per day, respectively). However, there was a significant difference between the two periods of treatment in the responders as for the mean daily number of beta 2 agonist inhalations (2.4 +/- 0.1 in the budesonide period as compared with 5.3 +/ 0.1 in the placebo period; p < 0.005). We conclude that treatment with inhaled steroids improved spirometry data and inhaled beta 2-agonist consumption in about 25% of patients with stable COPD, and this rate is increased to about 75% in patients who respond to beta 2-agonist inhalation. PMID- 7497763 TI - Noninvasive nasal mask ventilation beyond the ICU for an exacerbation of chronic respiratory insufficiency. AB - STUDY OBJECTIVE: To assess the usefulness of noninvasive nasal mask ventilation (NMV) in the treatment of an exacerbation of chronic respiratory insufficiency in patients stable enough to be admitted to a non-ICU ward. DESIGN: A prospective study in which the beneficial effect of NMV was compared with conservative treatment. SETTING: A ward of respiratory medicine of a tertiary-referral teaching hospital. PATIENTS: The study group included 15 patients with acute respiratory acidosis. These patients had pH less than 7.35 and PaCO2 more than 60 mm Hg, respiratory rate of 30 breaths or less per minute, hemodynamic stability, and alertness and willingness of cooperation with the NMV treatment. The control group consisted of 16 patients who fulfilled the same arterial blood gas requirements, retrospectively selected from the discharge forms of the ward of respiratory medicine for the year 1993. INTERVENTIONS: Patients underwent NMV for two sessions per day (one in the morning and one in the afternoon), each session lasting 4 h. A volumetric respirator (Monnal D; Taema; Paris, France) was used in four patients with restrictive disease. A positive-pressure ventilator (DP90; Taema; Paris, France) was used in 11 patients with obstructive disease. Control patients received standard medical, oxygen, and chest physical therapy. RESULTS: As compared with pre-NMV values, mean pH was significantly higher at 4 h of NMV after the patient's ventilatory adaptation (t = 8.814, p < 0.001) and at the end of NMV (t = 12.06, p < 0.001). Ventilatory support also produced a significant improvement in hypercapnia (pre-NMV vs NMV after the patient's ventilatory adaptation, t = 6.675, p < 0.001; pre-NMV vs post-NMV, t = 6.976, p < 0.001). Posttreatment pH and PaCO2 values were significantly higher and lower, respectively, in NMV-treated patients than in controls. At the end of treatment, a significantly higher PaO2/FIO2 ratio was documented in the study group than in controls (post-NMV vs posttreatment, t = 2.846, p < 0.01). CONCLUSIONS: NMV associated with standard treatment may be more beneficial than conservative treatment alone for improving blood gas exchange in patients with chronic respiratory insufficiency admitted to the hospital (but not the ICU) for an episode of acute decompensation and respiratory acidosis. PMID- 7497764 TI - CPAP machine performance and altitude. AB - STUDY RATIONALE AND OBJECTIVE: Sleep-disordered breathing is commonly treated with nasally applied continuous positive airway pressure (CPAP). Typically, pressures are titrated to pneumatically splint the airway to prevent its collapse in response to negative inspiratory pressure. This investigation was prompted by several patient complaints of sleep-related breathing difficulty associated with travel to high altitudes. CPAP devices create pressure with fan-generated airflow; therefore, CPAP performance should behave according to collective fan laws. MEASUREMENTS AND RESULTS: In the present study, we examined the effect of simulated altitude change on four commercially available CPAP machines. Machines were tested using anatomic airway mannequins in an altitude chamber. We made three simulated ascents to 12,000 feet with machines set at 5, 10, and 12 cm H2O sea level pressure equivalents. We measured pressure using water manometers at 2,000-foot increments during ascent and descent. Mask pressures varied systematically with changing altitude in three machines. One machine, equipped with a pressure regulation feature, maintained pressure within 1 mm H2O at all pressure and altitude combinations. CONCLUSIONS: Altitude significantly alters delivered pressure according to predictions made by the fan laws, unless a unit has pressure-compensating features. Clinicians should consider this factor when CPAP is prescribed for patients who live or travel to places located at significantly higher or lower elevations than the titration site. PMID- 7497765 TI - Home mechanical ventilation. Demographics and user perspectives. AB - Although the number of users of home mechanical ventilation (HMV) is increasing, information regarding the users' perceptions of the impact of HMV on their lives is incomplete. We administered an open-ended questionnaire to 98 (48 male, 50 female) HMV users aged (mean +/- SD) 47.4 +/- 19.5 years with COPD (9%), thoracic restrictive disease (43%), and neuromuscular disease (48%). At the time of survey, subjects had received HMV for 59.5 +/- 58.3 months. Fifty-three percent were ventilated electively. Ventilator use was continuous (18%), at night only (37%), or at night with occasional daytime use (45%). Twenty-seven (28%) individuals identified themselves as being totally independent while 32 (33%) rated themselves as partially dependent on caregiver assistance for daily activities. Only 37 (38%) considered that they had made an informed choice when first starting HMV whereas 54 (55%) did not. The impact of using a mechanical ventilator was believed to be overwhelmingly positive (87%). Positive comments regarding the impact of the ventilator on lifestyle were grouped in the following categories: life sustaining, facilitating mobility, and improving physical symptoms. Negative comments were grouped in the following categories: limiting mobility, equipment issues, and social implications. HMV users with a tracheostomy volunteered significantly fewer positive statements than those ventilated noninvasively (p < 0.05). Whereas 52 (53%) of users indicated that they had experienced initial difficulties in coping with the ventilator, only 11 (11%) identified difficulties at the time of the survey. PMID- 7497766 TI - Long-term health status and quality of life outcomes of lung transplant recipients. AB - STUDY OBJECTIVE: Health status and quality of life (QOL) in lung transplant candidates and recipients were compared to determine the impact of transplantation, and whether recipients experience continued improvements in the years after transplant surgery. DESIGN: Two patient groups, adult lung transplant candidates (n = 44) and adult lung transplant recipients (n = 54), completed self report QOL questionnaires. Eighteen of the 54 recipients completed QOL questionnaires on two occasions, about 18 months apart, after lung transplant. The questionnaire included the Medical Outcome Study Health Survey (MOS 20) that assesses six dimensions of health-related QOL: physical, role and social function, mental health, health perceptions and bodily pain, as well as a self report Karnofsky Index and other indicators of QOL. SETTING: University medical center transplant service. RESULTS: Compared with candidates, recipients reported higher levels of happiness and more satisfaction with their life and their health. They also reported better function on the Karnofsky Index and in every MOS 20 dimension (p < 0.0001), except pain. No significant differences were found in comparisons among recipients, grouped by the time since their transplant. Eighteen recipients completed two QOL questionnaires after transplant. No significant differences were found between the earlier (median, 11 months) and later (median, 29 months) QOL responses for this group overall. However, recipients (n = 5) who developed obliterative bronchiolitis between assessments showed decrements in their health-related QOL. Dimensions particularly affected were physical and social functioning and bodily pain. CONCLUSIONS: Dramatic improvements in health status and QOL occur after successful lung transplant and remain stable over time. Obliterative bronchiolitis results in notable QOL reductions. PMID- 7497767 TI - The cost-effectiveness of lung transplantation. A pilot study. University of Washington Medical Center Lung Transplant Study Group. AB - OBJECTIVE: Lung transplantation is one of the fastest-growing solid organ transplant procedures in the world, yet its cost-effectiveness is unknown. We compared the costs and outcomes of the first 25 patients who received lung transplants at the University of Washington with 24 patients currently on the lung transplant waiting list. DESIGN: Inpatient and outpatient charges were obtained from the hospital billing service and home health agencies. Quality adjusted life year scores (QALYs) were computed from the following: (1) utility scores obtained through standard gamble interviews, and (2) published survival data from an international lung transplant registry and from studies of patients on lung transplant waiting lists. RESULTS: Transplantation charges averaged $164,989 (median, $152,071). Average monthly charges posttransplant were $11,917 in year 1 and $4,525 thereafter, vs $3,395 for waiting-list patients. Posttransplant utility scores were significantly higher than waiting-list scores (0.80 vs 0.68; p < 0.001). Life expectancy was not greater for lung transplant vs waiting-list patients (5.89 vs 5.32 years; p > 0.05), although quality-adjusted life expectancy did improve significantly. After converting charges to costs, the incremental cost per QALY gained for posttransplant compared with waiting-list patients was $176,817. CONCLUSIONS: Lung transplantation is very expensive, although it can substantially improve quality of life. Two-thirds of care costs are incurred after transplantation. The principal barriers to cost-effectiveness at present are the high cost of postrecovery care and marginal gains in life expectancy compared with conservative care. PMID- 7497768 TI - Gastroparesis after lung transplantation. Potential role in postoperative respiratory complications. AB - BACKGROUND: We observed an unexpectedly high incidence of postoperative gastroparesis among lung and heart-lung transplant recipients. PURPOSE: To identify the incidence of GI complications and to describe the clinical profiles of patients who developed symptomatic gastroparesis after lung transplantation. PATIENTS AND METHODS: Retrospective study of GI symptoms and complications identified during 3 years of follow-up of 38 adult lung and heart-lung transplant recipients. RESULTS: Sixteen of 38 patients (42%) reported one or more GI complaint and received a specific GI diagnosis. Nine of 38 patients (24%) complained of early satiety, epigastric fullness, anorexia, nausea, or vomiting. Gastroparesis was suspected when endoscopic evaluation revealed undigested food in the stomach after overnight fast and symptoms could not be attributed to peptide disease or cytomegalovirus gastritis. Delayed gastric emptying was confirmed by gastric scintigraphy. Mean gastric empty (t1/2) was 263 +/- 115 min (normal < 95 min). Gastroparesis occurred in 4 of 13 right lung, 2 of 12 left lung, 1 of 9 bilateral single lung, and 2 of 4 heart-lung recipients (p = NS). Patients responded partially to metoclopramide or cisapride, with the exception of two patients who required placement of jejunal feeding tubes secondary to severe symptoms. In long-term follow-up, symptoms resolved in all patients and treatment with medications or mechanical intervention was successfully discontinued. Four of nine patients (44%) suffering from gastroparesis developed obliterative bronchiolitis (OB). Food particles were discovered in the BAL fluid of two such symptomatic patients. In contrast, only 6 of 29 (21%) nonsymptomatic patients developed OB (p = 0.16). CONCLUSION: Symptomatic gastroparesis is a frequent complication of lung or heart-lung transplantation that may promote microaspiration into the lung allograft. PMID- 7497769 TI - Bacteremic pneumococcal pneumonia. A community hospital experience. AB - We reviewed our experience with bacteremic pneumococcal pneumonia (BPP) over a 1 year period at a 600-bed community teaching hospital; 26 cases were identified. The mean age was 57.5 years and there were 12 male and 14 female subjects. Cough, sputum production, fever, and mental status changes were the most frequent symptoms. Risk factors included drug abuse in 10, HIV in 4, current smoking in 7, diabetes in 3, and cancer in 3. The mean PaO2/FIo2 ratio was 274. Radiographic features included a consolidation pattern in 7, bronchopneumonia in 15, combined in 1, and an initial normal film in 3. Average length of stay (LOS) was 11 days with an overall mortality of 11.5%. Four patients required mechanical ventilation, two meeting the criteria for ARDS (if this group were eliminated, LOS would be 8.4 days). Three of these survived. Four patients had organisms resistant to penicillin and all survived. We conclude that (1) BPP remains a serious but treatable infection particularly when utilizing full supportive care; (2) the bronchopneumonia x-ray film pattern was associated with all the mortality; and (3) the occurrence of penicillin resistance did not contribute to the mortality, since early recognition and the use of appropriate antibiotics saved all of these patients. PMID- 7497770 TI - Increased soluble CD14 levels in BAL fluid in pulmonary tuberculosis. AB - INTRODUCTION: CD14 is a cell surface glycoprotein expressed mainly on mature monocytes and macrophages. Soluble CD14 (sCD14) is present in normal plasma and is found increased in serum of patients with septicemia, polytrauma, and sarcoidosis. In active sarcoidosis, increased levels of sCD14 in BAL supernatant have been demonstrated. STUDY OBJECTIVE: To investigate sCD14 levels in BAL of pulmonary tuberculosis (PTB), another inflammatory disease characterized by granuloma formation. METHODS: BAL was performed in 12 patients with active but untreated PTB and 12 healthy persons. Cytologic and immunocytologic characteristics were determined. sCD14 was measured by a sandwich enzyme-linked immunosorbent assay. RESULTS: The level of sCD14 in patients with PTB was increased compared with controls (mean +/- SEM: 34.4 +/- 9.6 ng/mL vs 11.5 +/- 2.2 ng/mL; p = 0.02). No correlation was found between sCD14 levels and BAL cell differentials or lymphocyte surface markers. CONCLUSION: Similar to sarcoidosis, increased levels of sCD14 are found in BAL supernatant of PTB patients. PMID- 7497773 TI - Hypercapnic arousal responses in Prader-Willi syndrome. AB - STUDY OBJECTIVE: Prader-Willi syndrome (PWS) is characterized by a number of abnormalities of hypothalamic function, such as hyperphagia, short stature, temperature instability, hypogonadotropic hypogonadism, and neurosecretory growth hormone deficiency. Patients with PWS are reported to have sleep-disordered breathing and have blunted hypercapnic ventilatory responses secondary to abnormal peripheral chemoreceptor function. Thus, we hypothesized that hypercapnic arousal responses would be abnormal in PWS. DESIGN: Hypercapnic arousal responses were tested in ten nonobese children and adults with PWS, aged 17.7 +/- 2.5 (SEM) years, 70% female, and nine control subjects, aged 14.2 +/- 2.6 years, 67% female. Hypercapnic challenges were performed during stage 3/4 non rapid eye movement sleep. RESULTS: The PWS subjects had a significantly higher arousal threshold to hypercapnia compared with the controls (53 +/- 1.0 vs 46 +/- 1.7 mm Hg; p < 0.01). The PWS subjects had significantly higher baseline end tidal CO2 levels (42 +/- 0.8 vs 38 +/- 1.1 mm Hg; p < 0.01) and more central apneas greater than 15 s/h of sleep (1.5 +/- 0.3 vs 0.1 +/- 0.1; p < 0.01). CONCLUSIONS: Elevated hypercapnic arousal thresholds during sleep are found in PWS subjects; these may be a manifestation of abnormal peripheral chemoreceptor function and may further contribute to sleep-disordered breathing in PWS patients. PMID- 7497771 TI - Thoracic nodal staging with PET imaging with 18FDG in patients with bronchogenic carcinoma. AB - PURPOSE: To assess the role of positron emission tomographic (PET) imaging with 18-fluoro-2-deoxyglucose (18FDG) in detecting thoracic lymph node metastases in patients with bronchogenic carcinoma. MATERIALS AND METHODS: Over a 2-year period, any patient presenting to our institution with newly diagnosed bronchogenic carcinoma who was to have thoracic nodes sampled was considered eligible. All PET studies were performed prior to nodal sampling and areas of increased uptake were mapped according to the American Thoracic Society classification. Studies were correlated with CT and pathology. Sensitivity and specificity for predicting nodal metastases was calculated. RESULTS: Forty-two patients had 62 nodal stations (40 hilar/lobar, 22 mediastinal) sampled. The sensitivity and specificity for hilar/lobar lymph node station metastases using PET imaging was 73% and 76%, respectively. With CT, the sensitivity and specificity were 27% and 86%. The sensitivity and specificity using PET imaging for mediastinal node station metastases was 92% and 100%, respectively, while with CT the figures were 58% and 80%. The sensitivity and specificity for combined thoracic nodal station metastases using PET imaging was 83% and 82%, respectively, while with CT it was 43% and 85%. There was a strong statistical relationship between positive PET imaging and lymph node abnormalities. CONCLUSIONS: 18FDG-PET imaging is accurate in detecting thoracic lymph node metastases in patients with bronchogenic carcinoma. Normal results of PET studies virtually preclude the need for mediastinal nodal sampling prior to surgery, whereas abnormal results of studies most likely represent mediastinal metastases. Treatment can be based on the extent of disease suggested by PET imaging. PMID- 7497772 TI - Intrathoracic Kaposi's sarcoma. CT findings. AB - AIM: To describe the thin CT scans findings in AIDS patients with intrathoracic Kaposi's sarcoma (KS). MATERIAL AND METHODS: Fifty-three CT scans of patients with KS were retrospectively reviewed. The diagnosis of intrathoracic KS was established histologically (n = 17) or on the association of skin KS and the visualization of characteristic endobronchial lesions (n = 36). CT scans were performed with thin slices (2 mm) obtained at 10-mm intervals, and a 512 x 512 reconstruction matrix. No patients had Pneumocystis carinii pneumonia within the 3 months preceding the CT scan examination. RESULTS: Numerous nodules (n = 42), tumoral masses (n = 28), bronchovascular pathways thickening (n = 35), and pleural effusions (n = 28) were the most frequent patterns. Septal lines (n = 15), ground-glass opacities (n = 3), and mediastinal adenopathies (n = 8) were not frequent. CONCLUSION: Numerous nodules, tumoral masses, bronchovascular pathways thickening, and bilateral pleural effusions were the main signs of intrathoracic KS; their association (66%) is very characteristic. An opportunistic infection or mycobacteriosis must be sought if the thin CT scans reveal ground-glass opacities and/or mediastinal adenopathies. PMID- 7497775 TI - Acute epiglottitis. An 18-year experience in Rhode Island. AB - OBJECTIVE: To assess the incidence, clinical characteristics, management, and outcome of epiglottitis in a defined population over an 18-year period. DESIGN: Case series. SETTING: The state of Rhode Island, 1975 through 1992. PATIENTS OR OTHER PARTICIPANTS: Cases who met predetermined criteria for acute epiglottitis identified from hospital discharges and the State Medical Examiner's log of prehospitalization deaths. MAIN OUTCOME MEASURES: Incidence by year and age, clinical presentation, results of diagnostic evaluations, management, and outcome. RESULTS: Four hundred seven cases were identified, 134 in children and 273 in adults. Incidence in children dropped from 38 cases in the first 3 years of the study to 1 case in the last 3 years (p < 0.001). Adult cases increased from 17 in the first 3 years to 69 in the last 3 years (p < 0.001). Seventy-nine percent of adults and 32% of children were treated without an artificial airway. Factors associated with airway obstruction included symptomatic respiratory difficulty, stridor, drooling, shorter duration of symptoms, enlarged epiglottis on radiograph, and Haemophilus influenzae bacteremia (p < 0.001 for each). Twelve patients died (3 children and 9 adults), with all cases of fatal respiratory obstruction occurring within 12 h of presentation. CONCLUSIONS: There have been significant changes in the clinical epidemiology of epiglottitis, which now occurs almost exclusively in adults, often with less severe symptoms and a lower incidence of H influenzae infection. While careful observation is indicated for all patients, the data suggest that those with certain clinical characteristics can be treated safely without an immediate artificial airway. PMID- 7497774 TI - Reappraisal of distal diagnostic testing in the diagnosis of ICU-acquired pneumonia. AB - BACKGROUND: The thresholds of the diagnostic procedures performed to diagnose ICU acquired pneumonia (IAP) are either speculated or incompletely tested. PURPOSE: To evaluate the best threshold of protected specimen brush (PSB), plugged telescoping catheter (PTC), BAL culture (BAL C), and direct examination of cytocentrifugated lavage fluid (BAL D) to diagnose IAP. Each mechanically ventilated patient with suspected IAP underwent bronchoscopy successively with PSB, PTC, and BAL in the lung segment identified radiographically. POPULATION: One hundred twenty-two episodes of suspected IAP (occurring in 26% of all mechanically ventilated patients) were studied. Forty-five patients had definite IAP, and 58 had no IAP. Diagnosis was uncertain in 19 cases. RESULTS: Using the classic thresholds, sensitivity was 67% for PSB, 54% for PTC, 59% for BAL D, and 77% for BAL C. Specificity was 88% for PSB, 77% for PTC, 98% for BAL D, and 77% for BAL C. We used receiver operating characteristics methods to reappraise thresholds. Decreasing the thresholds to 500 cfu/mL for PSB, 10(2) cfu/mL for PTC, 2% cells containing bacteria for BAL D, 4 x 10(3) cfu/mL for BAL C increased the sensitivities (plus 14%, 23%, 25%, 10%, respectively) and moderately decreased the specificities (minus 4%, 9%, 2%, 4%, respectively) of the four examinations. The association of PSB with a 500 cfu/mL threshold and BAL D with a 2% threshold recovered all but one episode of pneumonia (SE 96 +/- 4%) with a 84 +/- 10% specificity. For a similar ICU population, these "best" thresholds increased negative predictive value with a minimal decrease of positive predictive value. They need to be confirmed in multiple ICU settings in prospective fashion. PMID- 7497777 TI - The effect of late-onset ventilator-associated pneumonia in determining patient mortality. AB - STUDY OBJECTIVE: To determine whether the development of late-onset ventilator associated pneumonia (VAP) is associated with an increased risk of hospital mortality. DESIGN: Prospective cohort study. SETTING: ICUs of two university affiliated teaching hospitals. PATIENTS: Three hundred fourteen patients admitted to an ICU who required mechanical ventilation for greater than 5 days. INTERVENTIONS: Prospective patient surveillance and data collection. MEASUREMENTS: The primary outcome measures were the development of late-onset VAP (ie, occurring > 96 h after intubation) and hospital mortality. RESULTS: Late onset VAP was observed in 87 patients (27.7%). Thirty-four (39.1%) patients with late-onset VAP died during hospitalization compared with 85 patients (37.4%) without late-onset VAP (relative risk, 1.04; 95% confidence interval [CI], 0.76 to 1.43). Twenty patients (6.4%) developed late-onset VAP due to a "high-risk" pathogen (ie, Pseudomonas aeruginosa, Acinetobacter sp, Xanthomonas maltophilia) with an associated mortality rate of 65%. Stepwise logistic regression analysis identified five variables as independent risk factors for hospital mortality (p < 0.05): an organ system failure index of 3 or greater (adjusted odds ratio [AOR], 3.4; 95% CI, 2.0 to 5.8; p < 0.001), having a nonsurgical diagnosis (AOR, 2.1; 95% CI, 1.3 to 3.6; p = 0.002), a premorbid lifestyle score of 2 or greater (AOR, 1.8; 95% CI, 1.1 to 2.9; p = 0.015), acquiring late-onset VAP due to a "high risk" pathogen (AOR, 3.4; 95% CI, 1.2 to 10.0; p = 0.025), and having received antacids or histamine type-2 receptor antagonists (AOR, 1.7; 95% CI, 1.0 to 2.9; p = 0.034). Additionally, we found the occurrence of late-onset VAP due to high risk pathogens to be the most important predictor of hospital mortality among patients developing VAP (AOR, 5.4; 95% CI, 2.8 to 10.3; p = 0.009). CONCLUSIONS: Nosocomial pneumonia due to certain high-risk microorganisms is an independent risk factor for hospital mortality among patients requiring prolonged mechanical ventilation. We suggest that future investigations of late-onset VAP stratify patient outcomes according to the distribution of high-risk pathogens when reporting their results. PMID- 7497776 TI - Increasing splanchnic blood flow in the critically III. AB - STUDY OBJECTIVE: To assess the effect of low dose dopexamine and dopamine on splanchnic blood flow as measured by gastric intramucosal pH, hepatic metabolism of lidocaine (lignocaine) to monoethylglycinexy-lidide (MEGX), and plasma disappearance rate of indocyanine green (ICG). DESIGN: Single-blind randomization of patients with a gastric intramucosal acidosis to receive dopexamine (ten patients), dopamine (ten patients), or saline solution (five control patients) for 2 h. SETTING: All 25 patients were in the ICU of Guys' Hospital. PATIENTS: All patients met the criteria for the diagnosis of the systemic inflammatory response syndrome, were mechanically ventilated, and had pulmonary artery catheters placed. All had a low gastric intramucosal pH and had a median first 24 h acute physiology and chronic health evaluation (II) score of 22 (range, 7 to 40). MEASUREMENTS AND INTERVENTIONS: Baseline measurements of gastric intramucosal pH, MEGX formation from lidocaine, ICG plasma disappearance rate, heart rate, mean arterial pressure, pulmonary artery occlusion pressure, cardiac index, oxygen delivery index, oxygen uptake index, systemic vascular resistance, and arterial pH were taken. Dopexamine (1 mg.kg-1.min-1), dopamine (2.5 mg.kg 1.min-1), or 0.9% saline solution was then infused for 2 h, after which a repeated set of the measurements was taken. RESULTS: Dopexamine at a low dose had no effect on any of the systemic measurements. The median intramucosal pH rose from 7.23 to 7.35 (p < 0.005), the median ICG plasma disappearance rate from 7.6 to 11.3%.min-1 (p < 0.02), and the median MEGX concentration from 4 to 10.2 ng.mL 1 (p < 0.005). Dopamine had no effect on any of the measured variables. There were no changes in the control group. CONCLUSIONS: Low-dose dopexamine increases splanchnic blood flow as measured by gastric intramucosal pH, MEGX formation from lidocaine, and ICG clearance. The lack of any change in the systemic measurements suggests that these effects are the result of a selective vasodilatation of the splanchnic vessels. At the dose used in this study, dopamine had no effect on splanchnic blood flow. Dopexamine may be useful in the management of splanchnic ischemia in the critically ill. PMID- 7497778 TI - Airway protection during experimental CPR. AB - BACKGROUND: Experimental studies recently demonstrated that positive pressure ventilation may not be essential for initial cardiopulmonary resuscitation. Nevertheless, oxygen enrichment of inspired gas mixtures and spontaneous gasping were associated with increased resuscitability and survival after cardiac arrest. However, as yet unresolved is the benefit of early airway control under conditions simulating "sudden death" due to ventricular fibrillation. METHODS: Twenty adult, male Sprague-Dawley rats were randomly assigned to one of two groups in which the airway was unprotected or protected by an oropharyngeal airway of our design. Cardiac arrest was induced by an alternating current delivered to the right ventricular endocardium. Oxygen was delivered to a hood that was loosely applied over the head of the each animal at a flow rate of 1 L/min. Precordial compression was initiated after 4 min of untreated ventricular fibrillation and defibrillation was attempted 6 min later. After spontaneous circulation had been restored, a tracheostomy was performed and the animals were mechanically ventilated with 100% oxygen for an additional interval of 1 h. Animals were then returned to their cages and observed for an additional 24 h. RESULTS: Spontaneous circulation was restored in each of the animals who had an oropharyngeal airway and nine of ten animals in the absence of an artificial airway. In each group, seven animals survived for more than 24 h. Animals in which the airway had been protected had significantly greater frequency of spontaneous gasping (28 +/- 13/min vs 13 +/- 9/min; p < 0.05) and significantly higher arterial oxygen saturation (77 +/- 19% vs vs 55 +/- 25%; p < 0.05). CONCLUSION: In the setting of experimental cardiac resuscitation, the insertion of an artificial airway increased the frequency of spontaneous gasping and arterial oxygenation. Nevertheless, no significant differences in resuscitability or postresuscitation survival were associated with insertion of the artificial airway. PMID- 7497779 TI - Metered-dose inhaler salbutamol-induced tracheal epithelial lesions in intubated rabbits. Effects of the site of administration and time. AB - STUDY OBJECTIVES: To determine (1) whether metered-dose inhaler (MDI) salbutamol administered at the elbow connector of the anesthetic circuit produced tracheal epithelial lesions in intubated rabbits, and (2) the time course for resolution of tracheal lesions produced by MDI salbutamol through an intratracheal catheter. DESIGN: Prospective, randomized, controlled trial. SETTING: University-affiliated animal research laboratory. PARTICIPANTS: Thirty-nine adult New Zealand white rabbits. INTERVENTIONS: (1) Twenty-one intubated rabbits received 0,5, or 20 puffs of MDI salbutamol delivered at the elbow connector of the anesthetic circuit. (2) Eighteen intubated rabbits received five puffs of MDI salbutamol through an intratracheal catheter and were killed 1 h, 24 h, or 1 week later. MEASUREMENTS: Samples of trachea, bronchi, and lungs were examined by light microscopy, and the degree of epithelial injury was assessed semiquantitatively. RESULTS: MDI salbutamol (5 or 20 puffs) administered at the elbow did not induce tracheal epithelial injury. When administered through an intratracheal catheter, MDI salbutamol (five puffs) produced moderate or severe tracheal epithelial injury in those killed 1 h after the study. Evidence of epithelial regeneration was observed 24 h after the injury and recovery was virtually complete by 1 week. CONCLUSION: Epithelial lesions do not occur when the MDI salbutamol (5 or 20 puffs) is administered at the elbow connector of the ventilation circuit. Tracheal epithelial lesions produced by MDI salbutamol (five puffs) administered through an intratracheal catheter resolve within 1 week of the injury. PMID- 7497780 TI - Blunt myocardial injury. PMID- 7497781 TI - Indications for valve surgery in asymptomatic patients with aortic and mitral stenosis. PMID- 7497782 TI - Thrombolytic therapy of left-sided prosthetic valve thrombosis. AB - Thrombosis of left-sided prosthetic valves is an uncommon yet potentially serious complication. Thrombolytic therapy has been proposed as an alternative to surgical methods in treating this condition. We sought to determine from a review of the literature what outcomes may be expected subsequent to thrombolytic administration and what groups may be at risk or benefit from this approach. We searched for studies including two or more patients treated with thrombolytic agents. Ten studies were reviewed. We describe and include two patients with valve thrombosis treated at our institution. A total of 182 episodes of prosthetic valve thrombosis in 162 patients were examined. Clinical success was achieved in 72.0% of cases and there was a mortality risk of 9.9%. Clinical success from thrombolysis was significantly related to the degree of heart failure at presentation and aortic valve position. Clinical success was not related to the duration of symptoms, time from valve replacement to obstruction, or valve type. Rethrombosis of successfully treated valve occurred subsequently in 19.5% of cases. Repeated thrombolytic administration in these patients was associated with similar rates of success as those treated for their initial episode of prosthetic valve thrombosis. Candidates for thrombolytic therapy include patients with obstructive valve thrombosis with or without congestive heart failure who are hemodynamically stable. Duration of time since valve replacement or symptom onset does not limit successful outcome. Close observation and aggressive maintenance of anticoagulation after therapy is suggested. PMID- 7497783 TI - Institutional control measures for tuberculosis in the era of multiple drug resistance. ACCP/ATS Consensus Conference. American College of Chest Physicians and the American Thoracic Society. PMID- 7497784 TI - Business and management training for physicians. CME or the third degree. PMID- 7497785 TI - Continuous extrapleural intercostal nerve block with continuous infusion of lidocaine after thoracotomy. A descriptive pilot study. AB - Continuous extrapleural intercostal block (EPIB) with bupivacaine has been reported to be an effective analgesic technique in patients after thoracotomy. We report a retrospective study of EPIB using a continuous infusion of 1% lidocaine hydrochloride at a dose of 1 mg/kg/h. A posterior parietal pleural pocket was created and cannulated with a 16-g polyethylene catheter. Lidocaine was perfused over a 3-day period following surgery. Patients also had access to morphine sulfate via patient-controlled analgesia. Eighteen consecutive posterolateral thoracotomies (in 17 patients) performed during a 6-month period were reviewed. Serum lidocaine exceeded the toxic level of 5 microgram/mL in only one patient, a 104.5-kg man who had a level of 5.9 micrograms/mL on postoperative day 2 but experienced no clinical toxicity. Pain was evaluated by verbal analog scores (0 = no pain and 10 = worst pain), which averaged 3.02, 3.14, and 2.8 in the 3 days following surgery. Mean total daily MS doses were 24.3, 37.75, and 34.32 mg (range, 0 to 94 mg). Sedation was scored on a 1 to 5 scale. Mean scores were 2.78, 2.56, and 2.6. No patient died or had a major respiratory complication. Continuous EPIB with lidocaine appears to be a promising adjuvant technique in the management of postthoracotomy pain. Effectiveness needs to be confirmed in a prospective randomized study. PMID- 7497786 TI - A case of pulmonary arteriovenous malformation in a patient with brain abscess successfully treated with video-assisted thoracoscopic resection. AB - A 45-year-old women was admitted to the hospital with a brain abscess due to asymptomatic pulmonary arteriovenous malformation (PAVM). The brain abscess was removed by craniotomy and excision following antibiotic therapy. The stapled wedge excision of the lung with the PAVM was successful under video-assisted thoracoscopic surgery. PMID- 7497788 TI - Persistent tachycardia. Diagnosis and management. PMID- 7497787 TI - Study of endoscopic staple formation by specimen radiography on pulmonary wedges. AB - The advent in video-assisted thoracic surgery has rendered us to rely more and more on mechanical devices. We prospectively studied staple formation on resected lung specimen by radiography and attempted to correlate this with the clinical outcome. From February 1994 to January 1995, 36 consecutive pulmonary wedges (23 apical bullae, 12 pulmonary metastases, 1 tuberculoma) from 31 patients who had undergone resection by endoscopic staple-cutter (Endo GIA30, USSC) were examined by two-plane radiography for staple alignment and closure. Imperfect staple formation was found in 21 (58%) of resected specimens. The median size of the resected bullae is 4.2 mL (range, 2.0 to 58.8 mL) compared with 36 mL (range, 1.2 to 222.8 mL) in the resected metastases and tuberculoma specimen. Despite detecting more imperfect staples in the metastases group (77% vs 57%), there was little difference in the two groups with respect to postoperative chest drainage duration (median, 3 vs 2 days) or hospital stay (median, 2 vs 2 days). We conclude that the currently available endoscopic staplers are by no means perfect, especially for resection of larger specimens, even though this may not be apparent clinically. A continuous search for improvement in endoscopic staplers designed specifically for lung resection is needed. PMID- 7497789 TI - Pressure waveform tracing. Constrictive pericarditis. PMID- 7497790 TI - Cavitating lung lesions and a cerebral mass. PMID- 7497792 TI - A late sternal wound infection caused by hematogenous spread of bacteria. AB - A 56-year-old man presented with a sternotomy wound infection 6 months after coronary artery bypass grafting. The organism responsible was group B beta hemolytic Streptococcus. This organism was simultaneously cultured from an infected diabetic ulcer on the patient's foot as well as from a total knee prosthesis. The Streptococcus apparently spread hematogenously to the sternum, an extremely rare cause of sternotomy wound infection. PMID- 7497791 TI - Sudden onset of large pericardial effusion in a 27-year-old man with AIDS. PMID- 7497794 TI - Multiple pulmonary air fluid levels as a presentation of congestive heart failure. AB - Congestive heart failure and emphysema are common diseases in western society. The presence of these diseases in a single patient can lead to diagnostic uncertainty. A case of congestive heart failure presenting as multiple air fluid levels within lung bullae is reported. PMID- 7497793 TI - Three-dimensional image reconstruction of partial anomalous pulmonary venous return to the superior vena cava. AB - A case of a rare partial anomalous pulmonary venous return of the right upper lobe into the superior vena cava is reported. Multiple three-dimensional image reconstructions in association with spiral CT are used in the aim of clarifying this abnormality of pulmonary venous drainage. PMID- 7497795 TI - Previously unrecognized intrapericardial hematoma leading to refractory abdominal ascites. AB - Delayed traumatic pericardial syndromes are well recognized. We describe a case in which a patient presented 3 years after an initial trauma with manifestations of constrictive pericarditis. The etiology in this report is attributable to a large intrapericardial hematoma, which is rarely described. This article also illustrates the complimentary nature of magnetic imaging and Doppler echocardiography in the evaluation of pericardial disease. PMID- 7497797 TI - Digital clubbing associated with polymyositis and interstitial lung disease. AB - Clubbing of the fingers is commonly associated with interstitial lung diseases (ILDs). Although ILD occurs in as many as 40% of patients with polymyositis/dermatomyositis (PM/DM), clubbing of the digits has never been reported to occur in patients with PM/DM and ILD. We report the first case of clubbing associated with PM/DM and ILD. PMID- 7497796 TI - Complete heart block and severe tricuspid regurgitation after radiotherapy. Case report and review of the literature. AB - Cardiac complications can occur long after chest radiotherapy. We describe a patient who developed both valve disease and complete heart block at different intervals following radiotherapy for Hodgkin's disease. The combined presentation of these two very rare cardiac complications and surgery for radiation-induced tricuspid valve disease have not been described before. PMID- 7497799 TI - Effect of prostacyclin on microvascular pressures in a patient with pulmonary veno-occlusive disease. AB - Continuous-infusion prostacyclin improves symptom scores and decreases mortality in patients with primary pulmonary hypertension, but use of prostacyclin in patients with pulmonary veno-occlusive disease may precipitate pulmonary edema. A patient with pulmonary veno-occlusive disease received a graduated intravenous infusion of prostacyclin and pulmonary capillary pressures were calculated during prostacyclin dose ranging. Calculated capillary pressure increased with low-dose prostacyclin (< or = 6 ng/kg/min) but decreased with higher doses. These data suggest that the post-capillary pulmonary venules in our patient had reversible vasomotor tone, but required a higher dose of prostacyclin to vasodilate than did the precapillary arterioles. PMID- 7497798 TI - Myocardial cysticercosis detected by ultrafast CT. AB - Cysticercosis can affect any organ of the body although central nervous system manifestations are the most common. Cysticercosis involving the myocardium is extremely rare and is usually diagnosed postmortem. We report a case of cysticercosis involving the myocardium diagnosed antemortem using ultrafast CT. PMID- 7497800 TI - Usual interstitial pneumonia associated with primary Sjogren's syndrome. AB - We describe the first case, to our knowledge, of usual interstitial pneumonia (UIP) as the pulmonary manifestation in primary Sjogren's syndrome (SjS). A 45 year-old woman was admitted to our hospital because of a dry cough and an interstitial shadow on a chest roentgenogram. Labial biopsy and sialogram confirmed a diagnosis of SjS. BAL fluid analysis revealed lymphocytosis with a decreased CD4/CD8 ratio compatible with bronchiolitis obliterans organizing pneumonia or lymphoid interstitial pneumonia. Open-lung biopsy specimen, however, showed evidence of UIP. Open-lung biopsy was a useful and necessary examination to determine the nature of the pulmonary complication in primary SjS. Conservative treatment without corticosteroids maintained a stable condition for a follow-up period of 3 years. PMID- 7497801 TI - Intermittent cocaine use associated with recurrent dissection of the thoracic and abdominal aorta. AB - Acute aortic dissection has been reported with the use of cocaine. We report a case of intermittent cocaine use spanning nearly 5 years and leading to recurrent dissection and extension of the false lumen. The patient repeatedly declined surgical correction. Management involved aggressive pharmacologic blood pressure control, close monitoring, and encouragement to enter drug rehabilitation. PMID- 7497802 TI - Emergency coronary revascularization in a liver transplant recipient. AB - A liver transplant recipient in cardiogenic shock from an acute myocardial infarction and failed coronary angioplasty underwent successful emergency coronary artery bypass graft surgery. Our case is only the fourth documented report of a coronary artery bypass graft in a liver transplant patient and the first conducted on an emergency basis in a patient in cardiogenic shock. Preservation of both cardiac and liver function has been evident in all cases. Based on these findings, liver transplantation should not be a deterrent or contraindication to coronary artery bypass or cardiac surgery. PMID- 7497803 TI - Thoracoscopic management of pleural effusions. PMID- 7497804 TI - Adenosine deaminase in pericardial fluids. Which test is best? PMID- 7497805 TI - Use of the Swan-Ganz catheter in a community hospital. PMID- 7497807 TI - Asthma mortality and beta-agonists. PMID- 7497806 TI - Criteria for pulmonary thromboendarterectomy. PMID- 7497808 TI - Unplanned extubations. PMID- 7497809 TI - Chest X-ray screening for lung cancer. PMID- 7497811 TI - Vocal cord dysfunction syndrome and "steroid-dependent" asthmatics. PMID- 7497810 TI - Is a pulmonary sequestration supplied by the coronary artery? PMID- 7497812 TI - Pulmonary hypertension and right heart failure. PMID- 7497813 TI - Is not the treatment program of dyspnea management strategies effective for COPD? PMID- 7497814 TI - Fulminant malignant arrythmia and multiorgan failure in acute arsenic poisoning. PMID- 7497815 TI - Chyloptyses found in a patient with lymphangioleiomyomatosis. PMID- 7497816 TI - Inhaling aerosolized Dakin's solution. PMID- 7497817 TI - Doxycycline activity against Streptococcus pneumoniae. PMID- 7497818 TI - Neonatal stress reactivity: predictions to later emotional temperament. AB - To investigate the relations among popular measures of neonatal stress and their link to subsequent temperament, 50 full-term newborns from a normal care nursery were examined responding to a heelstick blood draw. Baseline and heelstick measures of behavioral state, heart period, vagal tone, and salivary cortisol were obtained. Recovery measures of behavioral and cardiac activity were also analyzed. Mothers completed Rothbart's Infant Behavior Questionnaire when their infants reached 6 months of age. Baseline vagal tone predicted cortisol in response to the heelstick, suggesting that baseline vagal tone reflects the infants' ability to react to stressors. Greater reactivity to the heelstick (more crying, shorter heart periods, lower vagal tone, and higher cortisol) was associated with lower scores on "Distress-to-Limitations" temperament at 6 months. This finding was consistent with the expectation that the capacity to react strongly to an aversive stimulus would reflect better neurobehavioral organization in the newborn. Recovery measures of cardiac activity approximated and were correlated with baseline measures indicating the strong self-righting properties of the healthy newborn. Finally, vagal tone and salivary cortisol measures were not significantly related, suggesting the importance of assessing both systems in studies of the ontogeny of stress-temperament relations. PMID- 7497819 TI - Congruence between mothers' and fathers' differential treatment of siblings: links with family relations and children's well-being. AB - We studied patterns of mothers' and fathers' differential treatment of firstborn (average age 10.5 years) and secondborn (average age 8 years) school-age siblings, and we examined the links between parents' differential treatment and children's well-being and dyadic family relationships. Mothers, fathers, and both siblings in 110 families were interviewed in their homes. For each dimension of parental behavior that we assessed (i.e., differential affection and discipline) we created groups of families that reflected mothers' and fathers' levels of differential treatment (e.g., discipline the firstborn more, equal treatment, discipline the secondborn more). Although we detected substantial correspondence between the 2 parents' differential treatment, we found a sizable group of families in which parents' reports were incongruent (i.e., 1 parent reported equal and the other differential treatment). Parental patterns were linked to differences between the siblings' well-being and both sibling and parent-child relationships, with younger siblings exhibiting greater vulnerability to differential treatment. Incongruence in differential warmth was associated with marital distress. PMID- 7497820 TI - Helpfulness and the development of competence in adolescence. AB - Using data from a representative panel of 1,000 Minnesota youth, this paper explores "helpfulness" in 2 spheres of adolescents' lives: the home and paid work settings. We examine the social structural conditions under which helpful behaviors are elicited, the interrelations of helpfulness and competence across 2 years of middle adolescence, and whether social circumstances moderate the effects of helpfulness on competence. Both boys' and girls' helpfulness in the home is responsive to family need. Furthermore, helpfulness at work and girls' competence are reciprocally related. We find evidence that the effects of helpfulness depend on the helper's motivations and the act's meaning, as shaped by the social context. Girls' competence is diminished by helpfulness in the home under conditions of poor father-daughter relationships and coercive maternal control. PMID- 7497821 TI - Antisocial boys and their friends in early adolescence: relationship characteristics, quality, and interactional process. AB - This study examines the close friendships of early adolescent boys in relation to antisocial behavior. 186 13-14-year-old boys and their close friends were interviewed, assessed at school, and videotaped in a problem-solving task. Similarity was observed between the demographic characteristics and antisocial behavior of the boys and their close friends. There was a tendency for the close friends of antisocial boys to live within the same neighborhood block and to have met in unstructured, unsupervised activities. Direct observations of interactions with close friends revealed a reliable correlation between antisocial behavior, directives, and negative reciprocity. Positive interactions within the friendship were uncorrelated with antisocial behavior and relationship quality. Implications of these findings for clinical and developmental theory are discussed. PMID- 7497822 TI - Sex differences in neonatal stress reactivity. AB - The question of whether a sexually dimorphic stress reaction exists prior to extensive socialization was addressed by examining sex differences in physiological and behavioral stress reactivity, in healthy, term neonates, after a mildly stressful behavioral assessment procedure. The Neonatal Behavior Assessment Scale (NBAS; Brazelton, 1973) was administered to 18 male and 18 female neonates. Heart rate (HR), salivary cortisol, and behavioral states were assessed before and after the exam. Sex differences included higher cortisol response in males and a greater change in HR and higher NBAS motor performance cluster score in females. Salivary cortisol, HR change, NBAS cluster scores, and behavioral states after NBAS provided 100% discrimination between male and female infants. These findings suggest that there are neonatal sex differences in behavioral and physiological stress reactivity prior to socialization. PMID- 7497823 TI - Adolescents' and young adults' conceptions of civil liberties: freedom of speech and religion. AB - This study examined adolescents' and young adults' conceptions of freedom of speech and religion (civil liberties). 48 adolescents and young adults in 3 grade levels (mean ages 12-8, 16-10, and 19-6) were administered a structured interview containing assessments of civil liberties in general, in straightforward (unconflicted) applications, and in conflict with other social and moral concerns, including law, physical and psychological harm, and equality of opportunity. Freedom of speech and religion were conceptualized as universal rights and applied to social events in unconflicted contexts at all ages. A diverse array of rationales, differentiated according to type of freedom, were used at all ages to ground conceptions of universal freedoms. Judgments of civil liberties in conflicts exhibited several sources of variation, including developmental differences, situational or contextual variation determined by the particular types of issues in conflict, and individual differences. Results are consistent with the proposition that judgments of civil liberties reflect age related patterns of coordination of delimited social and moral concepts rather than general orientations. PMID- 7497824 TI - Moral reasoning about sexually transmitted diseases. AB - The purpose of this research was to investigate moral reasoning related to sexual behavior that could lead to the transmission of sexually transmitted diseases (STDs). Using hypothetical dilemmas about situations in which STDs can be transmitted, respondents were asked to explain why they believed the characters should or should not engage in risky behaviors. 40 college freshmen (M = 18.3 years) and 32 college seniors (M = 22.3 years) participated. Using Kohlberg's moral stage theory and Gilligan's moral orientation model, the interviews were scored for moral stage and moral orientation. Results indicated that the older age group had a significantly higher stage of moral reasoning than the younger age group when responding to dilemmas about STDs. There was a significant difference in moral stage between dilemmas, reflecting the possible effect of dilemma content on moral reasoning. The overall pattern of results shows nonsignificant gender differences in stage of moral reasoning and moral orientation. Clinical and theoretical implications of these findings for understanding the role of moral reasoning in sexual risky behavior are discussed. PMID- 7497825 TI - "I hardly cried when I got my shot!" Influencing children's reports about a visit to their pediatrician. AB - We examined, in 2 phases, the influence of postevent suggestions on children's reports of their visits to a pediatrician. Phase 1 examined the effect of giving one of 3 types of feedback to 5-year-old children immediately following their Diphtheria Pertussis Tetanus (DPT) inoculation. Children were given pain affirming feedback (the shot hurt), pain-denying feedback (the shot did not hurt), or neutral feedback (the shot is over). 1 week later, they did not differ in their reports concerning how much the shot hurt or how much they cried. In Phase 2, the same children were visited approximately 1 year after their inoculation. During 3 separate visits, they were either given additional pain denying or neutral feedback. They were also given misleading or nonmisleading information about the actions of the pediatrician and the assistant. Children given pain-denying feedback reported that they cried less and that the shot hurt less than did children given neutral feedback. Those who were given misleading information about the actions of the assistant and the pediatrician made more false allegations about their actions than did children who were not given this information. These results challenge the view that suggestibility effects are confined to peripheral, nonaction events; in this study children's reports about salient actions involving their own bodies in stressful conditions were influenced. PMID- 7497826 TI - Parents' socialization of children's injury prevention: description and some initial parameters. AB - In a year-long participant observation study of remediative action following actual injury, 61 8- and 9-year-old children and their 27-46-year-old mothers wrote records and reported on more than 1,000 minor injuries in branching biweekly interviews. Mothers reported that 80.1% of injuries received no parent initiated remediation, 14% received only a lecture, and less than 3% of injuries were followed by parental action. Children reported that 96.1% of their injuries were followed by no remediative action and recalled lectures after only 1.2% of injuries. Remediative action was related to type of child activity (e.g., unstructured play was followed by remediation more often than more purposive behavior) and to mother's affect (e.g., anger) and beliefs (e.g., that injury was the child's fault or due to rule violation). The parameters that influenced remediative consequences, and thus that may influence future injury, are discussed. PMID- 7497827 TI - Affective influences on startle in five-month-old infants: reactions to facial expressions of emotions. AB - Convergent methodologies from studies of fear-potentiated startle in animals and studies of affective modulation of reflex blinks in humans were adapted in order to investigate infants' sensitivity to affective information conveyed by facial expressions of emotion. While 5-month-old infants viewed photographic slides of faces posed in happy, neutral, or angry expressions, a brief acoustic noise burst was presented to elicit the blink component of human startle. Blink size was augmented during the viewing of angry expressions and reduced during happy expressions. Infants did not show marked changes in behavioral reactions to the positive, neutral, and negative slides, although motor activity was slightly reduced during negative slides. Results suggest that, by 5 months, infants react to affective information conveyed by unfamiliar human faces. Potential mechanisms mediating the influence of affective stimuli on reflex excitability are considered. PMID- 7497828 TI - A developmental investigation of inattentiveness and hyperactivity. AB - The development of inattentiveness and hyperactivity in middle childhood was investigated using a prospective longitudinal approach. Endogenous and exogenous predictors measured in infancy and in early and middle childhood were examined independently and in combination. In early childhood, quality of caregiving more powerfully predicted distractibility, an early precursor of hyperactivity, than did early biological or temperament factors. Caregiving and contextual factors together with early distractibility significantly predicted hyperactivity in middle childhood. While environmental variables also predicted hyperactivity in later elementary years, these factors did not improve the prediction beyond the influence of hyperactivity in early elementary years. The findings support a developmental view of the origins and course of hyperactivity in childhood, that is, that the emergence and persistence of AD/HD symptoms depend on developmental history along with current circumstances. PMID- 7497829 TI - Temperamental origins of child and adolescent behavior problems: from age three to age fifteen. AB - We assessed relations between early temperament and behavior problems across 12 years in an unselected sample of over 800 children. Temperament measures were drawn from behavior ratings made by examiners who observed children at ages 3, 5, 7, and 9. Factor analyses revealed 3 dimensions at each age: Lack of Control, Approach, and Sluggishness. Temperament dimensions at ages 3 and 5 were correlated in theoretically coherent ways with behavior problems that were independently evaluated by parents and teachers at ages 9 and 11, and by parents at ages 13 and 15. Lack of Control was more strongly associated with later externalizing behavior problems than with internalizing problems; Approach was associated with fewer internalizing problems among boys; and Sluggishness was weakly associated with both anxiety and inattention, especially among girls. Lack of Control and Sluggishness were also associated with fewer adolescent competencies. These results suggest that early temperament may have predictive specificity for the development of later psychopathology. PMID- 7497830 TI - Associations among adult attachment representations, maternal sensitivity, and infant-mother attachment in a sample of adolescent mothers. AB - Associations among adolescent attachment organization, maternal sensitivity, and infant attachment organization were examined prospectively in 74 teenaged mother infant dyads. Pregnant teenagers' attachment organizations predicted both sensitivity and infant-mother attachments. Mothers classified autonomous (F) in the prenatal period showed higher levels of sensitivity at both 3 and 9 months than mothers classified dismissing (Ds), preoccupied (E), or unresolved (U). Correspondence between maternal attachment (F vs. Ds/E/U) and infant attachment (secure [B] vs. avoidant [A]/resistant [C]/disorganized [D]) was observed in 58 of 74 (78%) dyads. Exact 4-group (Ds/E/F/U and A/B/C/D) agreement was observed in 50 of 74 (68%) families. In contrast, associations between maternal sensitivity and infant attachment were not significant, leading to questions about the processes that link attachment representations, maternal behavior, and infant attachment in adolescent mothers. PMID- 7497831 TI - It takes two to replicate: a mediational model for the impact of parents' stress on adolescent adjustment. AB - In this study of parental stress and adolescent adjustment, experiences of negative life events during the recent past were used to generate a measure of acute stress. In addition, multiple indicators based on reports from various informants were used to estimate latent constructs for parental depression, discipline practices, and adolescent adjustment. Employing 2 independent samples of families from 2 different regions of the country (rural Iowa and a medium sized city in Oregon), structural equation models were used to test the hypothesis that in intact families acute stress experienced by parents is linked to boys' adjustment (average age equaled 11.8 years in the Oregon sample, 12.7 years in the Iowa sample) through 2 different causal mechanisms. The findings showed that parental stress was related to adjustment through stress-related parental depression that is, in turn, correlated with disrupted discipline practices. Poor discipline appears to provide the direct link with developmental outcomes. The structural equation model (SEM) used to test the proposed mediational process was consistent with the data for mothers and boys from both the Oregon and the Iowa samples. The similarity in results was less clear for fathers and boys. Implications of these results for future replication studies are discussed. PMID- 7497832 TI - Interaction style, problem-solving behavior, and family problem-solving effectiveness. AB - We proposed that an individual's characteristic style of interaction will predict his or her problem-solving behavior and family problem-solving effectiveness. We test this hypothesis for mothers, fathers, and adolescent boys and girls (M age = 12.7 years) in 431 rural families using both warm and hostile interaction styles. One set of videotape coders observed a general family discussion and measured interaction style. A year later, another, independent set of coders observed a family problem-solving task. Family members reported family problem-solving effectiveness immediately following the problem-solving task. The results indicated that a hostile interaction style directly predicted destructive problem solving behavior and indirectly predicted family problem-solving effectiveness. A warm interaction style related directly to constructive problem-solving behavior and indirectly to family problem-solving effectiveness. PMID- 7497833 TI - Morbidity of subtotal colectomy in patients with severe ulcerative colitis unresponsive to cyclosporin. AB - PURPOSE: The aim of this study was to document the morbidity of urgent subtotal colectomy and ileostomy in patients with severe ulcerative colitis who failed cyclosporin treatment. METHODS: We reviewed the charts of patients with severe ulcerative colitis who did not respond to cyclosporin treatment and underwent urgent subtotal colectomy and Brooke ileostomy at two inflammatory bowel disease centers over the 12-month period ending April 1994. RESULTS: Fourteen patients (6 males; mean age, 34 years) required an urgent subtotal colectomy and Brooke ileostomy after failing treatment with cyclosporin. There were no deaths. Eight patients (57 percent) developed post-operative complications, which included ileus (3), deep vein thrombosis (2), wound infection (2), and partial dehiscence of rectal stump (1). Mean length of postoperative hospital stay was 8.8 days. CONCLUSIONS: These initial data suggest that cyclosporin treatment may not influence the safety of urgent surgical treatment in severe ulcerative colitis. PMID- 7497835 TI - Extended resections are beneficial for patients with locally advanced colorectal cancer. AB - PURPOSE: Locally advanced colorectal cancer often requires extended resection to radically remove all tumor. This is the only chance for cure in these patients, but a higher complication rate would be expected. To evaluate the overall benefit for the patient, this study assesses morbidity and mortality as well as long-term survival of patients who underwent extended resection for a T3-T4 carcinoma. METHODS: Two hundred twenty patients with locally advanced adenocarcinoma of the colorectum were included. One hundred fifty presented with a T3 and 70 with a T4 tumor. Eighty-three patients underwent extended resection. In 38 patients extended en bloc resection was performed because of inflammatory adherence mimicking infiltration. Thirty-three patients who underwent extended resections were over 70 years of age. There were no significant differences between the groups that underwent extended or nonextended resections in age, sex, stage, or grading. RESULTS: pT4 lesions were significantly more frequent in the extended resection group than in the nonextended resection group. Mean survival was 44 months after extended resections and 45 months after nonextended resections. In the extended resection group there was no significant difference in mean survival between pT3 and pT4 stage patients within 46 and 38 months, respectively. In patients who underwent nonextended resections, however, there was a significant difference in mean survival within 48 months for pT3 and 28 for pT4 patients (P < 0.05). Postoperative morbidity and mortality were comparable between the extended resection group and the non-extended resection group. The presence of residual tumor influenced prognosis of patients significantly; R0 resections fared significantly better than patients who underwent R1 or R2 resections (55 and 51 to 14/12 and 23/8 months) (P < 0.01). Nodal stage and International Union Against Cancer stage were also significant determinants of prognosis. After extended resections mean survival morbidity and 30-day mortality in patients more than 70 years was similar to those less than 70 years. CONCLUSION: Because extended resections can achieve comparable results in locally more advanced colorectal cancer as nonextended resections in less advanced cancer, an aggressive surgical approach is warranted. PMID- 7497836 TI - Sucrase-isomaltase is an independent prognostic marker for colorectal carcinoma. AB - PURPOSE: Expression of disaccharidase sucrase-isomaltase (SI) is significantly enhanced during neoplastic transformation of colonic epithelium. Our study was designed to determine whether expression of SI within primary tumors was significantly associated with survival in patients with colorectal carcinoma (CRC). METHODS: SI expression was analyzed by immunohistochemistry in paraffin sections from 182 Stage I to III CRC that had been resected for cure at the New England Deaconess Hospital between 1965 and 1977. Expression was scored as absent or present in 1 to 50 percent or more than 50 percent of tumor cells. Associations were explored among SI expression, other clinical or pathologic variables, and overall survival. The data set is mature, with 91 (56 percent) patients who had died of CRC at a median follow-up of 96 months. RESULTS: Fifty five percent of primary CRC expressed SI. When the multivariate Cox analysis was performed, nodal status, T stage, primary site, grade, and SI expression were independent covariates. SI expression was not associated with the expression of other clinicopathologic variables but increased the risk of death from colorectal carcinoma by 1.83-fold. DISCUSSION: These results indicate that SI is a prognostic marker for CRC that is independent of stage-related variables in patients who have undergone potentially curative resections. PMID- 7497834 TI - Long-term results of anterior resection using the double-stapling technique. AB - PURPOSE: This study was designed to determine the anastomotic leak rate, local recurrence rate, and survival of patients undergoing anterior resection with the double-stapling technique for rectal cancer. METHODS: Between 1981 and 1992, 189 patients underwent a curative (166) or palliative (23) anterior resection using the double-stapling technique. A chart review was performed, and follow-up information was obtained from the patient or family physician. Follow-up was complete in 186 patients (98 percent). RESULTS: There were five (2.6 percent) stapler-related complications, of which two patients required a defunctioning colostomy. Postoperative mortality was 1.6 percent, and clinical leak rate was 7.3 percent. Clinical leak rate was significantly higher in patients with lesions in the lower third (20 percent) compared with those in the middle and upper thirds (9 and 1 percent, respectively; P < 0.05). After a mean follow-up of 32 +/ 29 months, local recurrence rate was 9.1 percent but was significantly higher in patients more than 65 years old (14 vs. 1 percent; P < 0.005) and in patients with resection margins less than 2.0 cm (17 vs. 5.5 percent; P < 0.05). Five-year survival was 78 percent. CONCLUSIONS: Anterior resection performed with the double-stapling technique has an acceptable clinical leak rate, local recurrence rate, and survival rate. However, the clinical leak rate appears to be increased in patients with low tumors and, therefore, a defunctioning colostomy should be considered. Resection margins of more than 2 cm are necessary. PMID- 7497837 TI - Ambulatory hemorrhoidectomy with CO2 laser. AB - PURPOSE: This study was undertaken to evaluate ambulatory hemorrhoidectomy using the CO2 laser. METHODS: Ninety consecutive patients (50 males, 40 females), 80 percent of whom had second or third degree hemorrhoids, had ambulatory hemorrhoidectomy performed with a CO2 laser in the left lateral Sims position under local anesthesia and intravenous sedation. Dissection was entirely performed with the CO2 laser using an open technique. RESULTS: One patient with polycythemia vera was admitted subsequently for secondary hemorrhage. All but three patients had healed within four weeks. No long-term sequelae were encountered. CONCLUSION: These results compare favorably with those obtained by others and show that ambulatory laser hemorrhoidectomy may simplify management in selected patients. PMID- 7497838 TI - Accuracy of serum C-reactive protein measurements in diagnosis of acute appendicitis compared with surgeon's clinical impression. AB - Diagnosis of acute appendicitis is established generally by the surgeon's clinical impression. Today, negative laparotomy rate because of clinical diagnosis is still 15 to 25 percent. PURPOSE: This study was designed to determine the accuracy of C-reactive protein (CRP) measurements in the diagnosis of acute appendicitis and to compare it with the surgeon's clinical diagnosis. METHODS: One hundred eight consecutive patients were studied prospectively. Depending on results of the examination by a surgeon, patients underwent surgery for acute appendicitis. Serum CRP measurements were performed before the operations but were not taken into account for the decision of laparotomy to compare it with the surgeon's clinical diagnosis. RESULTS: Histopathologic findings confirmed acute appendicitis in 90 patients. Normal appendixes were removed in the remaining 18 patients. Mean serum CRP value was 5 (range, 0-12.6) mg/l in patients with normal appendix, 33.8 (range, 5-85.1) mg/l in patients with nonperforated appendicitis, and 128.5 (range, 79.2-230) mg/l in patients with perforated appendixes. These differences were highly significant (P < 10(-6)). Serum CRP levels were normal in three patients with acute appendicitis. Thus, the false-negative rate of CRP was 3 percent. Of 18 patients with normal appendectomy serum CRP levels were slightly elevated in two patients. We determined, therefore, a false-positive rate of CRP as 11 percent. CRP levels were false negative in three patients and false-positive in two patients. Thus, CRP levels were true (positive or negative) in the remaining 103 patients. On the other hand, the diagnosis depending on surgeon's clinical impression was true in 90 patients and false in 18 patients. This difference was statistically significant (P = 0.0035). In the present study the sensitivity, specificity, and accuracy of serum CRP measurements were calculated as 93.5, 80, and 91 percent, respectively. CONCLUSION: We found that elevated serum CRP levels support surgeon's clinical diagnosis. We recommend CRP measurement as a routine laboratory test in patients with suspected diagnosis of acute appendicitis. PMID- 7497839 TI - Manometric asymmetry in the anal canal in controls and patients with fecal incontinence. AB - PURPOSE: This study was designed to ascertain the existence of manometric asymmetry in the anal canal in controls and in patients with fecal incontinence. METHODS: A computerized manometric technique with an eight-channel perfusion catheter was used. Coefficient of variation (CV, a mathematical expression of the degree or magnitude of deviation of a set of data points from the mean) was developed in this study as an index of anal canal manometric asymmetry. RESULTS: At 1 cm from the anal verge, mean CV was 9.3 and 8.7 percent in asymptomatic control males and females, respectively. During a maximum voluntary squeeze effort, mean CV in control males was 9.3 percent and was not significantly different (P = 0.28) from that in control females (7.8 percent). In 14 consecutive female patients with fecal incontinence, mean CV at rest (1 cm from the anal verge) was 21 percent and was significantly higher (P < 0.01) than in control females. Similarly, during a maximum voluntary squeeze effort, mean CV in patients with fecal incontinence was 20.5 percent and was significantly higher (P < 0.01) than in the female control group (7.8 percent). CONCLUSION: It is concluded that CV, a method of expressing anal sphincter manometric asymmetry, is a useful manometric parameter in the assessment of anorectal function. PMID- 7497840 TI - Latency measurement of rectoanal reflexes. AB - PURPOSE: Latency values of rectoanal reflexes may be altered in disorders of the pelvic floor. Evaluation of this relatively uninvestigated aspect of rectoanal reflexes may have diagnostic implications in patients with disorders of defecation. METHODS: We studied the latency of rectoanal inhibitory and excitatory reflexes to sequential balloon distention of the rectum with 60 ml and 120 ml of air in 14 normal controls (mean age, 41.5 (range, 19-66) years), in 14 patients with fecal incontinence (FI) (mean age, 44.2 (range, 28-72) years), and in 14 patients with slow transit constipation (STC) (mean age, 40.6 (range 22-68) years). RESULTS: The mean latency of inhibition (FI = 5.3 seconds; STC = 4.6 seconds; controls = 5.1 seconds) was remarkably similar for the three groups (P = 0.19). The mean latency of excitation in the proximal anal canal (FI = 2.8 seconds; STC = 2.5 seconds; controls = 2.8 seconds) was comparable in the three groups (P = 0.58). The mean latency of excitation in the distal anal canal (FI = 4.8 seconds; STC = 2.6 seconds; controls = 2.7 seconds) was prolonged in patients who were incontinent compared with the other two groups (P < 0.01). CONCLUSIONS: Proximal rectoanal excitation and inhibitory reflexes, when present, have a constant latency, irrespective of the underlying condition. The different latency values for proximal and distal rectoanal excitatory reflexes in patients with FI may indicate disparate denervation damage to the external anal sphincter. PMID- 7497842 TI - Does peritoneal mobilization increase laparoscopic acidosis? AB - PURPOSE: It was hypothesized that laparoscopic colon surgery may be associated with increased absorption of CO2 resulting from mobilization of lateral peritoneal reflections. METHODS: Six pigs underwent laparotomy with removal of a measured quadrant of peritoneum before insufflation to 15 mmHg with CO2. Six paired control animals also underwent insufflation with CO2 to 15 mmHg. Measurements of the end-tidal CO2 (PetCO2), arterial blood gas analysis for CO2 (PaCO2), and pH were performed before insufflation, at 5 and 10 minutes following insufflation, then at successive 15-minute intervals for a total of two hours, and 15 minutes following desufflation. No attempt was made to correct the hypercarbia by increasing minute ventilation. RESULTS: PaCO2 reached its maximum level at two hours with values of 70.77 +/- 5.54 mmHg and 64.62 +/- 5.18 mmHg in the peritonectomized and control groups, respectively. PetCO2 also peaked at two hours to 60 +/- 13 mmHg for the study group and 54 +/- 11 mmHg for controls. pH reached its nadir at two hours, falling from a baseline of 7.45 +/- 0.08 to 7.23 +/- 0.09 in the study group, and from 7.42 +/- 0.04 to 7.24 +/- 0.04 in the control group. There were no statistically significant differences between the two groups for any of the parameters measured at each time interval. CONCLUSIONS: The hypothesis that peritonectomy produces greater CO2 absorption during CO2 pneumoperitoneum was disproved under these experimental conditions. PMID- 7497841 TI - Management of early invasive colorectal cancer. Risk of recurrence and clinical guidelines. AB - PURPOSE: The purpose of this study was the evaluation of various factors in the formulation of guidelines for treatment of early invasive colorectal cancer, in which malignant cells extend through the muscularis mucosa into the submucosa but do not deeply invade the muscularis propria. METHOD: A total of 182 patients were followed for at least five years or until death, with early invasive cancer diagnosed between 1982 and 1989. Patients were grouped according to the level of invasion, as follows: 64 patients with slight carcinoma invasion of the muscularis mucosa (200-300 microns; sm1), 82 with intermediate invasion (sm2), and 36 with carcinoma invasion extending to the inner surface of the muscularis propria (sm3). RESULT: The configuration, diameter, and histologic grade of adenocarcinoma and lymphovascular invasion were correlated with level of invasion. After endoscopic polypectomy or local resection, 4 patients showed local recurrence and 13 patients showed lymph node metastasis. None of these 17 patients had sm1 disease. The level of invasion, configuration, and location were significant risk factors for development of lymph node metastasis or local recurrence (P < 0.05), but lymphovascular invasion, histologic grade, and diameter were not risk factors. CONCLUSIONS: Preoperative assessment of the level of invasion using this classification, in which the submucosa is divided into three depths, may decrease the incidence of unnecessary surgery for sessile polyps. Assessment according to the level of invasion is useful in the formulation of appropriate guidelines for the treatment of early invasive cancer. PMID- 7497843 TI - Subclinical time span of inflammatory bowel disease in patients with primary sclerosing cholangitis. AB - PURPOSE: This study is designed to describe colonic histology in patients with primary sclerosing cholangitis (PSC) without clinical symptoms of inflammatory bowel disease (IBD) and to do a follow-up study of these patients to find the time span from first detection of histologic signs until development of clinical symptoms of IBD. METHODS: In a cohort of 76 patients with PSC treated at Huddinge University Hospital, 11 patients did not have any clinical symptoms of IBD at the time of PSC diagnosis. Nine of these patients underwent diagnostic colonoscopy with multiple biopsies. RESULTS: In the group of nine PSC patients, without clinical signs of IBD undergoing colonoscopy, histologic signs of IBD were found in seven patients (6 ulcerative colitis and 1 Crohn's disease). Among them one had dysplasia, and another had epithelial changes probably positive for dysplasia. Two other patients had histologic signs of inflammation, however, not fully compatible with IBD. Three of 11 patients developed clinical symptoms of IBD after one, three, and seven years of follow-up since diagnostic colonoscopy. CONCLUSIONS: In patients with PSC, histologic signs of IBD, including premalignant changes, may precede development of clinical symptoms of IBD by as much as seven years. This indicates that IBD onset may have a substantial subclinical phase of IBD far longer than previously appreciated. This finding may be of clinical importance because underestimation of disease duration may delay inclusion of PSC patients with extensive colitis in colonoscopic surveillance programs. The subclinical phase may also allow the studies of early pathogenesis in vivo. PMID- 7497845 TI - Toxicity of nonsteroidal anti-inflammatory drugs in the large intestine. AB - PURPOSE: Adverse effects of nonsteroidal anti-inflammatory drugs (NSAIDS) on the upper gastrointestinal (GI) tract and small intestine are well described. Evidence is also accumulating that implicate NSAIDS in inducing and exacerbating damage in the distal GI tract. The purpose of this review is to identify possible adverse effects of NSAIDS on the large intestine and increase the clinical awareness of these toxicologic effects. METHODS: A literature review identified the diversity of toxicologic effects induced by NSAIDS in the large intestine. The epidemiology, pathogenesis, and clinical implications of these adverse effects are described. RESULTS: NSAID use has been associated with colonic bleeding, iron deficiency anemia, strictures, ulcerations, perforations, diarrhea, and death. In addition, NSAIDS can exacerbate inflammatory bowel disease and ulcerative colitis. The prevalence of NSAID-induced large intestinal damage is unknown. Diagnosis can be made by colonoscopy and barium scans. Although the clinical presentation of NSAID-induced gastropathy and enteropathy, bleeding or perforation, may be more dramatic than colonopathy, the overall clinical significance of these adverse effects of NSAIDS on the large intestine has not been fully characterized. CONCLUSIONS: This review illustrates that NSAID induced large bowel toxicity can cause significant morbidity in some patients, ranging from profuse diarrhea, chronic blood loss, and iron deficiency anemia to fatality. The pathogenesis is likely multifactorial and is thought to be related to inhibition of prostaglandin synthesis. Because NSAIDS are widely prescribed and some are available without a prescription, heightened awareness of these toxicologic manifestations throughout the GI tract may reduce morbidity. PMID- 7497844 TI - Serum selenium and colonic neoplastic risk. AB - BACKGROUND: Selenium deficiency has been associated with cancer risk in several organs. This association was investigated in neoplasia of the colorectum. DESIGN: A case-control study is reported with two patient series, colorectal cancer and colorectal adenomatous polyps, and a control group found to be free of colorectal neoplasia. Diagnosis was determined by colonoscopy and histologic review of suspected neoplasms. Serum drawn at the time of colonoscopy was subsequently assayed for selenium content, and quartiles based on selenium were defined. Crude and adjusted odds ratios with 95 percent confidence intervals for adenoma related to selenium were calculated, controlling for known or suspected risk factors including gender, age, race, body mass index, family history, tobacco use, alcohol consumption, serum beta carotene, serum alpha tocopherol, and serum ferritin. RESULTS: There were 138 controls who had no neoplastic disease, 139 adenoma patients, and 25 cancer patients. For adenoma, comparing higher quartiles of selenium to the first (lowest selenium), the adjusted odds ratio for the second quartile was 1.7 (95 percent confidence interval, 0.8-3.7), the third quartile was 1.4 (0.7-3.2), and the fourth (highest selenium) quartile was 1.8 (0.9-4). The odds ratios for cancer patients were 0.8 for the second quartile, 1 for the third quartile, and 1.7 for the fourth quartile. CONCLUSION: No trend could be detected toward a protective effect of higher levels of serum selenium for colonic benign or malignant tumors. PMID- 7497846 TI - Ligation device assisting low anterior anastomosis. AB - The use of the circular stapler has enabled surgeons to perform very low anterior resections where previously abdominoperineal proctectomy would be performed for carcinoma of the rectum. The use of the roticulator or other linear stapler devices has facilitated these anastomoses and precluded the necessity for pursestring sutures, thereby reducing the incidence of incomplete tissue rings, which should reduce the risk of anastomotic leaks. One significant drawback to the roticulator, especially in the narrow male pelvis, is its application for very low lesions. In many cases the stapler cannot be forced deeply enough into the pelvis to allow for a sufficient margin below the tumor. PURPOSE: The purpose of this article is to report results of a prototype that applies a gasket that automatically fixes the rectum to the rod of the circular stapler. METHOD: To date the device has been used 22 times in 14 patients (8 patients had both limbs attached using the device, whereas 6 patients had only the rectal stump attached in this manner). RESULTS: All anastomoses healed without complication, all patients had complete donuts of tissue, and only two anastomoses had to be reinforced. CONCLUSION: The use of this device may facilitate stapling of very low anastomoses for carcinoma of the rectum. PMID- 7497847 TI - Double-barreled wet colostomy. A simple method of urinary diversion for patients undergoing pelvic exenteration. AB - An easy and simple method for constructing a urinary diversion in patients undergoing pelvic exenteration for advanced rectal cancer is described. This procedure features double-barreled colostomy with a segment of the sigmoid colon 8 to 10 cm distal to the stoma as a urinary conduit. PMID- 7497848 TI - Isolated sigmoid tuberculosis. Report of a case. AB - PURPOSE: To heighten awareness of colonic tuberculosis (TB) as a once rare disease that is undergoing a resurgence in the United States. METHODS: Report of a case of isolated sigmoid tuberculosis with a brief literature review of the topic. RESULTS: TB can no longer be considered a rare disease in the United States because, in part, of the acquired immunodeficiency syndrome epidemic and because, in part, of increased immigration and lack of containment. The signs and symptoms of colonic TB are nonspecific; therefore, a high index of suspicion must be maintained. Only 20 percent of patients will have associated active pulmonary TB. Colonoscopy with multiple biopsies at ulcer margins should be performed for diagnosis. Tissue should be sent for routine histology and culture and smeared for direct visualization of acid-fast bacilli. If colonic TB is suspected, empiric treatment is warranted, despite negative histology, smear, and culture results. Patients will usually show a dramatic response in one to two weeks. Treatment is solely medical, and all patients should receive a full course of antituberculous chemotherapy. Exploratory laparotomy is necessary if diagnosis is in doubt, when there is concern about a neoplasm, or for complications including perforation, obstruction, hemorrhage, or fistulization. CONCLUSION: An increased awareness of intestinal TB coupled with familiarity of the pathophysiology, diagnostic methods, and treatment should increase the number of cases correctly diagnosed preoperatively and, therefore, improve the outcome of patients with this disease. PMID- 7497849 TI - Stimulated gracilis neosphincter--not as good as previously thought. Report of four cases. AB - PURPOSE: We report the outcome of four patients who had stimulated gracilis neosphincter for fecal incontinence to highlight functional problems, particularly in patients with impaired rectal evacuation. METHODS: The gracilis neosphincter operation consisted of a three-stage procedure in four patients with intractable incontinence, three of whom had had a pelvic floor repair. RESULTS: Despite successful muscle transposition and nerve stimulation, only one of four patients has a functioning neosphincter. One patient could not tolerate stimulation, and two were unable to evacuate the rectum. All three now have stomas, and even the functioning neosphincter patient requires regular bisacodyl (Dulcolax; CIBA Consumer Pharmaceuticals, Woodbridge, NJ) suppositories to achieve evacuation. CONCLUSION: The neosphincter is a successful sphincter but has no role for patients who cannot evacuate from the rectum. PMID- 7497850 TI - Comparison of body size measurements as predictors of NIDDM in Pima Indians. AB - OBJECTIVE--To determine and compare the abilities of various anthropometric measurements to predict the development of non-insulin-dependent diabetes mellitus (NIDDM) in Pima Indian men and women. RESEARCH DESIGN AND METHODS--A total of 290 male and 443 female Pima Indians were followed for up to 6 years for the development of NIDDM. A proportional hazards analysis was used to assess the ability of anthropometric measurements evaluated at baseline to predict NIDDM. Receiver operating characteristic (ROC) curves were used to compare individual variables in predicting NIDDM. RESULTS--In separate models controlled for age and sex, body mass index (BMI), waist circumference, thigh circumference, waist-to thigh ratio (WTR), weight, and percentage body fat (PBF) estimated by bioelectric resistance each predicted NIDDM, which developed in 30 men and 52 women. The highest incidence rate ratios (IRRs; for 1 SD of a variable) were for WTR in men and for PBF in women, although the confidence interval (CI) for PBF was wide. In stepwise analyses, WTR was the most significant predictor in men (IRR for 1 SD = 1.58, 95% CI = 1.20-2.07), and BMI was the most significant predictor in women (IRR for 1 SD = 1.65, 95% CI = 1.29-2.11). However, by ROC analyses, thigh circumference was the only variable significantly worse than WTR in men or BMI in women at predicting NIDDM. CONCLUSIONS-- Measurements such as waist circumference, WTR, weight, and BMI may be useful as more complicated measurements, such as PBF by bioelectrical resistance, for identifying groups of individuals whose body habitus places them at high risk of developing NIDDM. PMID- 7497851 TI - Kinetics of HbA1c, glycated albumin, and fructosamine and analysis of their weight functions against preceding plasma glucose level. AB - OBJECTIVE: To examine the kinetics of HbA1c, glycated albumin (GA), and fructosamine (FA) levels in response to plasma glucose change and their relationship with the preceding plasma glucose level. RESEARCH DESIGN AND METHODS: The time courses of HbA1c, GA, and FA after acute glycemic normalization were observed in nine patients with newly diagnosed non-insulin-dependent diabetes mellitus and compared with theoretical ones. Their weight functions against preceding plasma glucose level were analyzed assuming a stepwise plasma glucose change and compared with the theoretical prediction. RESULTS: The fasting plasma glucose level was acutely normalized after admission with a half-time of 6.3 +/- 2.4 days (mean +/- SD). The HbA1c level decreased linearly during the initial 2 months with a half-time of 34.6 +/- 10.1 days, followed by a gradual decrease thereafter. GA and FA levels decreased very rapidly during the initial 2 3 weeks with half-times of 17.1 +/- 2.8 and 12.2 +/- 4.8 days, respectively, followed by a gradual decrease thereafter. The time courses of HbA1c, GA, and FA agreed well with theoretically estimated decay curves. Experimental values of weight functions against the preceding plasma glucose level agreed well with the theoretical prediction. The weight functions for glycated proteins had maximum values on the days just before the measurement of glycated proteins and gradually decreased with an increasing time interval. The lengths of the periods over which the weight functions for HbA1c, GA, and FA extend back were estimated to be roughly 100, 40, and 30 days, respectively. CONCLUSIONS: The levels of HbA1c, GA, and FA do not reflect the simple mean but reflect the weighted mean of the preceding plasma glucose level over a considerably longer period than was previously speculated. PMID- 7497852 TI - Sex difference in lifestyle factors predictive of diabetes in Mexican-Americans. AB - OBJECTIVE: Little is known about the role of lifestyle factors in non-insulin dependent diabetes mellitus (NIDDM) incidence among Mexican-Americans. Therefore, we examined whether baseline lifestyle factors predictive of 8-year NIDDM incidence differ in Mexican-American men and women. RESEARCH DESIGN AND METHODS: We studied 353 Mexican-American men and 491 Mexican-American women free of diabetes at baseline who participated in the San Antonio Heart Study follow-up. Lifestyle factors examined were body mass index (BMI), energy intake (total calories/kg), grams of alcohol consumed per week, efforts to control weight by dieting and exercise, leisure physical activity, sugar avoidance, saturated fat/cholesterol avoidance, and 24-h dietary recall assessment of total calories and percentage of calories from total carbohydrate, sucrose, and starch and from total, saturated, monounsaturated, and polyunsaturated fat. Incidence of NIDDM was regressed on lifestyle factors separately for men and women using a backward elimination procedure. RESULTS: Lifestyle factors significantly associated with NIDDM incidence differed for the two sexes. In men, leisure physical activity, was inversely associated and alcohol consumption, weight control by dieting, and BMI were positively associated with NIDDM. In women, BMI was positively associated with NIDDM and was the strongest lifestyle predictor. Sugar avoidance and leisure physical activity were also associated with increased NIDDM risk, while weight control by dieting was associated with decreased NIDDM risk. Saturated fat/cholesterol avoidance, grams of alcohol consumed per week, and energy intake were also negatively and indirectly associated with NIDDM in women by means of their direct effects on BMI. CONCLUSIONS: It may be important to tailor interventions designed to prevent NIDDM in Mexican-Americans to address sex differences in lifestyle precursors of this disease PMID- 7497853 TI - Glucose, insulin, and glucagon levels during exercise in pancreas transplant recipients. AB - OBJECTIVE: To determine whether pancreas transplant recipients maintain normal blood glucose levels during physical exercise. RESEARCH DESIGN AND METHODS: We measured serum glucose, insulin, C-peptide, and plasma glucagon levels in six pancreas transplant recipients and six healthy control subjects matched for age, sex, body size, and level of conditioning during 1 h of bicycle exercise at a workload set to achieve 40% of each individual's previously determined peak oxygen consumption (VO2). RESULTS: Serum glucose values were not different between control subjects and transplant recipients before the start of exercise (5.0 +/- 0.1 and 4.9 +/-0.1 mmol/l, respectively). Serum glucose levels fell slightly but significantly in both recipients and control subjects during exercise. There were no significant differences in glucose levels between the two groups at any time point during exercise, although mean nadir glucose during exercise was slightly lower in transplant recipients compared with control subjects (4.4 +/- 0.1 vs. 4.8 +/- 0.1 mmol/l, P = 0.04). In control subjects, insulin and C-peptide levels fell significantly within 15-30 min of exercise and glucagon levels rose significantly after 60 min of exercise. In transplant recipients, there was a trend for insulin and C-peptide levels to fall and glucagon levels to rise during exercise, although these changes were delayed and were not statistically significant. CONCLUSIONS: No significant abnormalities in blood glucose were detected in pancreas transplant recipients during bicycle exercise at 40% of peak VO2 for 1 h. Compared with levels in control subjects, subtle alterations in insulin and glucagon levels may occur in transplant recipients during exercise. However, these alterations do not appear to result in either hyperglycemia or hypoglycemia during light exercise for up to 1 h. PMID- 7497854 TI - Screening for diabetes mellitus in adults. The utility of random capillary blood glucose measurements. AB - OBJECTIVE: Because half of the people with non-insulin-dependent diabetes mellitus (NIDDM) are undiagnosed and because near-normal glycemic control can prevent diabetic complications, we evaluated the use of field-based random capillary blood glucose measurement as a screening test for NIDDM. RESEARCH DESIGN AND METHODS: A cross-sectional sample of 828 Egyptians > or = 20 years of age underwent both a random capillary blood glucose measurement performed with a portable reflectance meter in the field and an oral glucose tolerance test in the laboratory. The sensitivity and specificity of random capillary blood glucose measurements in predicting the presence of NIDDM were evaluated. RESULTS: Multivariate analyses showed that the screening test performed better when subjects had eaten shortly before the test (area under receiver operating characteristic curve, 0.87 for a 1-h postprandial period compared with 0.69 for an 8-h postprandial period) and that the optimal capillary blood glucose cutoff points to define a positive test increased with age. For a postprandial period of 1 h, cutoff points of 115 mg/dl for individuals 30 years of age and 140 mg/dl for those 75 years of age yielded similar performance characteristics (sensitivity 82% and specificity 78% for those 30 years old; sensitivity 81% and specificity 80% for those 75 years old). CONCLUSIONS: Adjusting random capillary blood glucose measurements for the postprandial period and using age-specific cutoff point values can improve performance of the screening test. PMID- 7497855 TI - Effects of maternal gestational diabetes and adiposity on neonatal adiposity and blood pressure. AB - OBJECTIVE: To determine the effects of maternal factors, including prepregnancy maternal adiposity, weight gain during pregnancy, degree of abnormality of the glucose tolerance test, glycemia during pregnancy, and treatment with insulin versus diet therapy, on neonatal body weight, adiposity, and blood pressure in infants of mothers with gestational diabetes (IGDM) and control patients. RESEARCH DESIGN AND METHODS: A total of 119 term IGDM, including 57 large-for gestational-age (LGA) and 62 appropriate-for-gestational-age (AGA) infants, and 143 term control infants, including 74 LGA and 69 AGA infants, were prospectively enrolled. Maternal measurements of prepregnancy weight, height, and weight gain were abstracted from medical records. A diagnosis of gestational diabetes was made on the basis of an initial 1-h 50-g glucose screen value > or = 130 mg/dl followed by two abnormal values in a 100-g oral glucose tolerance test. Infant anthropometric measurements were obtained, and blood pressure was measured on day 2 of life. Correlation analyses and multiple regression analyses were performed to assess the relationships among maternal factors and neonatal adiposity and blood pressure. RESULTS: Multiple regression analyses to determine the effects of significant maternal factors on infant body mass index (BMI) revealed that prepregnancy weight and weight gain were significant predictors for both IGDM and control infants. An increased glucose screen predicted BMI for control subjects, whereas the mean 2nd and 3rd trimester glucose values were the significant predictors for IGDM. Also, increased newborn triceps skinfold thickness measurements correlated with increased systolic blood pressure for IGDM (r = 0.29, P < 0.03). CONCLUSIONS: Increased maternal prepregnancy weight, weight gain in pregnancy, and glycemia in pregnancy all place IGDM at increased risk of macrosomia and adiposity. Increased adiposity in the IGDM appears to be related to increased infant blood pressure. Longitudinal evaluation is needed to determine whether neonatal adiposity in IGDM is predictive of increased adiposity and blood pressure during childhood. PMID- 7497856 TI - High risk of IDDM in African-American and Hispanic children in Chicago, 1985 1990. AB - OBJECTIVE: To determine the incidence of insulin-dependent diabetes mellitus (IDDM) among African-American and Hispanic children < 18 years of age in the city of Chicago. These minority communities are large and heterogeneous with respect to socioeconomic status, length of time since migration, and place of origin, so that correlates of IDDM risk can be examined with precision. RESEARCH DESIGN AND METHODS: Cases occurring during the years 1985-1990 were drawn from records at 37 hospitals in Cook County. African-American and Hispanic patients using insulin, residing within the city limits, and < 18 years old at onset were included. Three secondary sources of cases were used: medical records of clinics associated with the Chicago Department of Health, a survey of unaffiliated neighborhood clinics, and lists of children attending diabetes camps. Overall ascertainment was estimated at 86%. RESULTS: There were 413 new cases during this 6-year interval. The average incidence of IDDM was 12.0/100,000 annually among African-American males, 12.1 among African-American females, 9.1 among Hispanic males, and 10.2 among Hispanic females. Mean age at onset was 11.1, 11.0, 10.7, and 10.1 years for African-American males, African-American females, Hispanic males, and Hispanic females, respectively. Fewer cases occurred during the summer months. Diabetes among the first-degree relatives of children from both ethnic groups was commonly noted on the medical charts. CONCLUSIONS: The incidence rates in Chicago fall near the upper limits of reports for both African-origin and Hispanic populations worldwide. The relatively early age at onset may point to an environmental factor associated with this high incidence of the disease. Further studies will provide valuable data on comorbid conditions, unusual diabetic syndromes, and family dynamics in childhood chronic disease. PMID- 7497857 TI - Diabetes in urban African-Americans. I. Cessation of insulin therapy is the major precipitating cause of diabetic ketoacidosis. AB - OBJECTIVE: To identify the causes of diabetic ketoacidosis (DKA) in a large urban hospital. RESEARCH DESIGN AND METHODS: Consecutive patients admitted during a 3 month period with a primary diagnosis of DKA and who had moderate-to-severe illness as shown by serum glucose > 13.9 mmol/l ( > 250 mg/dl), bicarbonate < 15 mmol/l, and pH < 7.35 were studied. Diabetes nurse educators interviewed patients and reviewed their medical records for the following: precipitating causes of DKA; content of previous diabetes education; frequency of blood glucose monitoring; recognition of symptoms of metabolic decompensation; and types of medical assistance obtained once patients were ill. RESULTS: There were 56 episodes of DKA, and 75% of the episodes were in patients with known diabetes. In the known diabetic patients, the most common cause of DKA was stopping insulin therapy, which occurred in 67% of episodes. Half of the patients (50%) stopped insulin because of reported lack of money to buy insulin from an outside pharmacy or get transportation to the hospital; 21% stopped insulin because of lack of appetite; 14% stopped insulin because of behavioral or psychological reasons; and 14% did so because they did not know how to manage diabetes on sick days. Of the known diabetic patients, > 80% recalled having been instructed as to blood glucose testing and acute and chronic complications, but fewer patients recalled having been instructed as to insulin dose adjustment (28%) or sick day management (35%). Symptoms of decompensated diabetes were recognized in 55% of the 42 episodes of DKA in patients with known diabetes. However, only 5% of patients contacted the Diabetes Unit when they became ill; the majority (95%) went directly to the emergency room. CONCLUSIONS: DKA occurred most often in patients with known diabetes who stopped insulin therapy because of reported lack of money for purchasing insulin or for transportation to the hospital and limited self care skills in diabetes management. In urban African-American populations, up to two-thirds of the episodes of DKA may be preventable by improving patient education and access to care. PMID- 7497858 TI - Relationship between habitual physical activity and insulin area among individuals with impaired glucose tolerance. The San Luis Valley Diabetes Study. AB - OBJECTIVE: To determine whether higher levels of physical activity are associated with lower fasting insulin levels and lower insulin areas under the oral glucose tolerance curve in individuals with impaired glucose tolerance (IGT) in a community setting. RESEARCH DESIGN AND METHODS: Data from a cross-sectional study of a population consisting of 219 Hispanic and non-Hispanic white men and women with IGT (by World Health Organization criteria) in two rural Colorado counties were analyzed. Total physical activity was assessed by a 7-day physical activity recall, from which metabolic equivalents (METs) were estimated (expressed as MET h/week). Relationships of MET h/week with fasting insulin levels and insulin areas were assessed while considering obesity, age, and other risk factors known to influence fasting insulin level and insulin area. RESULTS: Among all subjects, univariate analyses showed that higher physical activity levels were associated with lower mean insulin areas and fasting insulin levels (both P < 0.05). Multiple linear regression showed that higher levels of physical activity were significantly associated with lower values of of the insulin area (P < 0.001) but not with fasting insulin levels. The relationship between insulin area and habitual physical activity was independent of obesity, fat distribution, and age. CONCLUSIONS: On the basis of cross-sectional data, we conclude that higher levels of habitual physical activity are associated with lower insulin areas in a population of individuals with IGT. Understanding the impact of physical activity on markers of insulin action in individuals with IGT is important because of the greatly enhanced risk of non-insulin-dependent diabetes mellitus and, hence, cardiovascular disease in this population. PMID- 7497859 TI - Immunogenicity of long-term intraperitoneal insulin administration with implantable programmable pumps. Metabolic consequences. AB - OBJECTIVE: To assess immunogenicity of intraperitoneal insulin infusion via implanted pumps by two methods and to evaluate the possible influence of an increased antibody level on metabolic and clinical parameters. RESEARCH DESIGN AND METHODS: We studied insulin antibody levels in 17 type I diabetic patients before and until 24 months after implantation of a programmable pump delivering insulin intraperitoneally. Antibody levels were determined by radioimmunoassay (RIA) and enzyme-linked immunosorbent assay (ELISA). They were correlated with HbA1c, insulin requirements, free insulin, and the incidence of hypoglycemia. RESULTS: Insulin antibodies increased as soon as the 3rd month after implantation. This increase was sustained throughout the study period (month 0, 25.4 +/- 16.2%; month 3, 41.2 +/- 23.5%; month 12, 45.9 +/- 26%; month 24, 48.7 +/- 25%). The data was correlated with the two assay methods (RIA and ELISA). Postimplantation level was correlated with preimplantation level, which could indicate a predictive value of the latter . No correlation was observed with any metabolic parameters, particularly the number of hypoglycemic episodes. CONCLUSIONS: Our results indicate that intraperitoneal insulin administration by implantable programmable pumps leads to an increase of insulin antibodies, which are probably high-affinity antibodies (recognized by both RIA and ELISA). This increase in insulin immunogenicity did not induce significant metabolic consequences, which is reassuring for the future of programmable insulin pumps. PMID- 7497860 TI - Apolipoprotein E polymorphism as a risk factor for vascular disease in diabetic patients. AB - OBJECTIVE: To examine the prevalence of cardiovascular disease in diabetic patients as a function of apolipoprotein (apo) E polymorphism. RESEARCH DESIGN AND METHODS: The apo E phenotypes and plasma lipid, lipoprotein, and apo levels were determined for 517 Italian diabetic patients. The prevalence of cardiovascular disease (defined as ischemic heart disease [HD] and/or peripheral vascular disease and/or cerebrovascular disease) was assessed as a function of apo E polymorphism at entry and after 4 years. RESULTS: The occurrence of vascular disease did not differ significantly between diabetic patients in the various categories of apo E phenotype either at entry into the study or after 4 years. When expressed as a percentage of patients with disease, we observed--for E2, E3, and E4 carriers, respectively--at entry: IHD, 20.0% (n = 14), 21.0% (n = 79), and 21.5% (n = 14); and macroangiopathy, 24.3% (n = 17), 29.3% (n = 110), and 24.6% (n = 16). Apo E polymorphism did not make a significant contribution to multiple logistic regression models designed to identify the factors associated with the occurrence of vascular disease in diabetic patients. CONCLUSION: Apo E polymorphism and, notably, the apo E4 allele cannot be universally considered as a particular risk factor for cardiovascular disease in diabetic patients. PMID- 7497861 TI - Risk factors for lower extremity amputation in persons with diabetes. AB - OBJECTIVE: To determine the predictors of lower extremity amputation (LEA) in a cohort of persons with diabetes (primarily non-insulin-dependent). RESEARCH DESIGN AND METHODS: We conducted a nested case-control study among 10,068 patients from a large health maintenance organization who reported a diagnosis of diabetes at a multiphasic health checkup (MHC) (baseline) between 1964 and 1984. Average length of follow-up after baseline was 13.2 years. Case patients were 150 cohort members with a first, nontraumatic LEA after baseline. Control subjects were 278 cohort members who did not experience an LEA during follow-up, matched to patients on age, sex, and year of baseline MHC. The presence of diabetes at baseline was verified by chart review for both patients and control subjects. Cardiovascular disease risk factors were obtained at the baseline MHC; glucose control, other diabetes-related variables, preventive services, and other complications were obtained from chart review. RESULTS: Level of glucose control (P < 0.0001), duration of diabetes (P = 0.04), and baseline systolic blood pressure (P = 0.004) were independent predictors of amputation, as were microvascular complications (retinopathy, neuropathy, and nephropathy). History of stroke, but not myocardial infarction, was also independently predictive; type of diabetes, cigarette smoking, and total cholesterol level were not. Being African-American was unrelated to amputation risk in univariate or multivariate analyses in this insured population. CONCLUSIONS: LEA shares a risk factor profile with other microvascular complication of diabetes. Thus, control of blood glucose and blood pressure should reduce risk for amputation. African-Americans do not appear to be at increased risk for diabetes-related amputation when access to medical care is comparable. PMID- 7497862 TI - Reduced awareness of hypoglycemia in adults with IDDM. A prospective study of hypoglycemic frequency and associated symptoms. AB - OBJECTIVE: To prospectively evaluate the frequency and severity of hypoglycemic episodes in IDDM subjects who declare themselves to have reduced awareness of hypoglycemia, to validate their self-designations in their natural environment, and to determine objectively the presence or absence of autonomic and neuroglycopenic symptoms associated with their low blood glucose (BG) levels. RESEARCH DESIGN AND METHODS: A total of 78 insulin-dependent diabetes mellitus (IDDM) subjects (mean age 38.3 +/- 9.2 years; duration of diabetes 19.3 +/- 10.4 years) completed two sets of assessments separated by 6 months. The assessments included reports of frequency and severity of low BG, symptoms associated with low BG, and a BG symptom/estimation trial using a hand-held computer (HHC). Diaries of hypoglycemic episodes were kept for the intervening 6 months. HbA1 levels were determined at each assessment. RESULTS: Of the subjects, 39 declared themselves as having reduced awareness of hypoglycemia (reduced-awareness subjects). There were no differences between these reduced-awareness subjects and aware subjects with regard to age, sex, disease duration, insulin dose, or HbA1. During the HHC trials, reduced-awareness subjects were significantly less accurate in detecting BG < 3.9 mmol/l (33.2 +/- 47 vs. 47.6 +/- 50% detection, P = 0.001) and had significantly fewer autonomic (0.41 +/- 0.82 vs. 1.08 +/- 1.22, P = 0.006, reduced-awareness vs. aware) and neuroglycopenic (0.44 +/- 0.85 vs. 1.18 +/- 1.32, P = 0.004, reduced-awareness vs. aware) symptoms per subject. Prospective diary records revealed that reduced-awareness subjects experienced more moderate (351 vs. 238, P = 0.026) and severe (50 vs. 17, P = 0.0062) hypoglycemic events. The second assessment results were similar to the first and verified the reliability of the data. CONCLUSIONS: IDDM subjects who believe they have reduced awareness of hypoglycemia are generally correct. They have a history of more moderate and severe hypoglycemia, are less accurate at detecting BG < 3.9 mmol/l, and prospectively experience more moderate and severe hypoglycemia than do aware subjects. Neither disease duration nor level of glucose control explains their reduced awareness of hypoglycemia. Reduced-awareness individuals may benefit from interventions designed to teach them to recognize all of their potential early warning symptoms. PMID- 7497863 TI - A multicenter evaluation of blood glucose awareness training-II. AB - OBJECTIVE: Blood glucose awareness training (BGAT) teaches individuals with insulin-dependent diabetes to more accurately estimate/detect their blood glucose (BG) fluctuations. It has not, however, consistently resulted in improved ability to detect low BG. To assess an enhanced version of BGAT (BGAT-II), with more focus on increasing sensitivity to low BG events, a multicenter study was undertaken. Following up on previous findings that BGAT is most effective with individuals who are least accurate in estimating BG, this study explicitly recruited subjects who did and did not report reduced awareness of hypoglycemia. RESEARCH DESIGN AND METHODS: Seventy-eight subjects from three research sites participated in a repeated baseline design. Subjects' BG estimation accuracy and BG profiles were assessed 6 months before, immediately before, and immediately after BGAT-II. RESULTS: Post-treatment, BGAT-II led to better overall accuracy in detecting BG fluctuations and better detection of both low and high BG levels. This was achieved while the number of low readings of self-monitoring of blood glucose (SMBG) was reduced. Reduction in the number of low SMBG events was significant only for subjects reporting awareness of hypoglycemia. Detection of low BG was significant only for subjects reporting reduced awareness of hypoglycemia. Both groups demonstrated equivalent improvements in detection of high BG levels. CONCLUSIONS: BGAT may be an effective behavioral strategy for reversing hypoglycemic unawareness and an adjunct to intensive insulin therapy to reduce the occurrence of severe hypoglycemia. PMID- 7497866 TI - Incidence of IDDM in the province of Rzeszow, Poland, 0- to 29-year-old age group, 1980-1992. AB - OBJECTIVE: To study the incidence of insulin-dependent diabetes mellitus (IDDM) in the province of Rzeszow, Poland, and to test for differences in incidence by age at diagnosis, time cluster, sex, urban-rural population, and season. RESEARCH DESIGN AND METHODS: A registry was established in 1990 to collect new IDDM cases among people 0-29 years of age, from 1980 to 1992. Data were collected according to the Diabetes Epidemiology Research International Group recommendations. RESULTS: The average age-standardized incidence of IDDM was 5.31/100,000 population (95% confidence interval [CI] 4.67-6.04) for those 0-29 years of age. Among males, it was 5.68/100,000 (CI 4.77-6.77), and among females it was 4.93/100,000 (CI 4.07-5.97). The incidence for children 0-14 years old was 5.11/100,000 (CI 4.27-6.11) and for those 15-29 years old, 5.55/100,000 (CI 4.60 6.68). The incidence of IDDM in autumn (September through November) was the highest (87 of 243 cases) and differed statistically from each of the other seasons. The incidence of IDDM among the urban and rural populations did not differ. There were also no differences between the sexes and the groups divided by 5-year age intervals. CONCLUSIONS: This study documents the low incidence rate and seasonal dependence of IDDM among people 0-29 years of age in the province of Rzeszow, Poland. PMID- 7497864 TI - Determinants of 24-h blood pressure in IDDM patients. AB - OBJECTIVE: To assess the influence of basic clinical data on ambulatory blood pressure (AMBP) in insulin-dependent diabetes mellitus (IDDM) patients and to evaluate the reproducibility of the method. RESEARCH DESIGN AND METHODS: AMBP was measured in 66 IDDM patients with urinary albumin excretion (UAE) < 20 micrograms/min and in 53 healthy subjects. Determinants of AMBP were identified in a stepwise multiple regression model. In addition, 14 diabetic patients were monitored on two days of the same type, and 14 patients were monitored on one work day and one day off. RESULTS: In healthy subjects, sex was the most important determinant of 24-h blood pressure (BP), whereas UAE and age were the main covariates in diabetes patients. The male-female difference in 24-h diastolic BP (dBP) was 5.6 mmHg lower in diabetic patients than in healthy control subjects (P < 0.05). In patients with long diabetes duration, nighttime dBP (69 +/- 7 mmHg) was higher than in patients with medium diabetes duration (63 +/- 8 mmHg, P < 0.05; after matching for age and sex). Daytime dBP was 5 mmHg higher on a work day than on a day off (P < 0.02). Standard deviation of the difference for repeated measurement of 24-h systolic/diastolic BP in the same subject was 5.7/2.5 mmHg. CONCLUSIONS: The male-female difference in 24-h dBP was attenuated in diabetes. The influence of UAE on AMBP was noticed even in normoalbuminuric diabetic patients. Standardized AMBP was highly reproducible. PMID- 7497865 TI - Tolrestat in the primary prevention of diabetic neuropathy. AB - OBJECTIVE: To compare the effects of tolrestat and placebo in patients with subclinical diabetic neuropathy. RESEARCH DESIGN AND METHODS: Non-insulin dependent diabetes mellitus (NIDDM) patients with early involvement of the autonomic nervous system were identified by only one pathological (outside the 99% confidence interval of the normal population) squatting test (vagal or sympathetic). Fifty-seven patients entered a randomized, placebo-controlled, double-blind, parallel 52-week study of tolrestat at a dose of 200 mg/day. Cardiovascular reflex tests (squatting vagal and sympathetic tests, pressure gain, deep breathing, lying-to-standing, Valsalva maneuver, and orthostatic hypertension), vibration thresholds, tendon reflexes, and muscle strength were assessed throughout the study. RESULTS: At 12 months, nerve function significantly improved in patients receiving tolrestat and deteriorated in patients taking placebo. At baseline, the squatting vagal test was normal in 16 patients in the tolrestat group and in 15 patients in the placebo group. At 12 months, 25 patients taking tolrestat had a normalized squatting test, but only 6 patients taking placebo did (P = 0.02). Vibration perception threshold improved by a value of 6 +/- 3 V in the tolrestat group (P < 0.001) and deteriorated by a value of 3 +/- 1.8 V (P < 0.001) in the placebo group. CONCLUSIONS: Tolrestat may be useful in the primary prevention of diabetic neuropathy. PMID- 7497867 TI - Prevalence of diabetes and IGT in Yonchon County, South Korea. AB - OBJECTIVE: To determine the prevalence of diabetes and impaired glucose tolerance (IGT) in Yonchon County of South Korea and to investigate their associated factors. RESEARCH DESIGN AND METHODS: We performed a population-based cross sectional study with random cluster sampling of residents > or = 30 years of age. Among the 3,804 residents sampled, a total of 2,520 participants had a standard 75-g oral glucose tolerance test and answered a detailed questionnaire. We also collected standard anthropometric data. RESULTS: If the data for participants in the age range of 30-64 years were adjusted to the standard world population, the prevalence of diabetes was 7.2% and the prevalence of IGT was 8.9%. It was observed that the significant factors associated with diabetes were waist-to-hip circumference ratio, serum triglyceride levels, age, systolic blood pressure, family history of diabetes, and locality. CONCLUSIONS: The prevalence of diabetes in Yonchon County was substantially higher than was previously suggested. The risk of diabetes increased with the increased central obesity and metabolic disturbances associated with insulin resistance. PMID- 7497869 TI - National data on the epidemiology of IDDM in Greece. Cases diagnosed in 1992. Hellenic Epidemiology Study Group. AB - OBJECTIVE: To detect the incidence, geographic distribution, and seasonal variation of insulin-dependent diabetes mellitus (IDDM) in all of Greece. RESEARCH DESIGN AND METHODS: One hundred and thirty-seven children (75 boys and 62 girls) up to 14.9 years of age were diagnosed with IDDM in Greece in 1992. RESULTS: The age-adjusted incidence of IDDM in Greece in 1992 was 6.25/100,000 population (boys: 6.17/100,000;: girls: 6.28/100,000). There was a significant pubertal peak. The incidence of IDDM was significantly higher in metropolitan (10.0/100,000) than in rural (4.21/100,000), semirural (7.48/100,000), or urban areas (5.36/100,000) of Athens. There was a marginal difference between rural and semirural areas. No significant seasonal variation in the incidence of IDDM was observed. CONCLUSIONS: We conclude that the incidence of IDDM in Greece is among the five lowest in Europe. An impressive clustering of cases was observed in the Athens metropolitan area compared with the rural and semirural areas, an observation that possibly underlines the importance of environmental factors (stressful situations, infections, dietary habits, and others) in the expression of IDDM. PMID- 7497870 TI - Prevalence of diabetes and hypertension in a rural population of Bangladesh. AB - OBJECTIVE--To determine the prevalence of non-insulin-dependent diabetes mellitus (NIDDM), impaired glucose tolerance (IGT), and hypertension in a rural community of Bangladesh. RESEARCH DESIGN AND METHODS--A cluster sampling of 1,005 subjects > 15 years of age in the rural community of Dohar was investigated. Capillary blood glucose of fasting and 2 h after 75 g oral glucose (2hBG) were estimated. World Health Organization criteria were used for diagnosis of NIDDM and IGT. Blood pressure, height, and weight were also measured. RESULTS--The crude prevalence of NIDDM was 2.1% (men 3.1, women 1.3%) and IGT was 13.3% (men 14.4, women 12.4%). Age-adjusted (30-64 years of age) prevalence was 2.23% (95% confidence interval [CI] 1.01-3.45) for NIDDM and 15.67% (95% CI 12.59-18.75) for IGT. Prevalence of hypertension with systolic blood pressure (sBP) > or = 140 mmHg was 10.5% and with diastolic blood pressure (dBP) > 90 mmHg was 9.0%. Increased age was the risk factor for NIDDM, IGT, and hypertension; whereas increased BMI showed inconsistent association with them. Relative risk for sBP with higher BMI ( < 22.0 vs. > or = 22.1) was 1.94 with CI 1.55-2.43 and for dBP it was 2.2 with CI 1.40-3.46. Correlation of sBP was significant with age, BMI, and 2hBG. Similar correlation was also observed with dBP. CONCLUSIONS--High prevalences of NIDDM, IGT, and hypertension were observed among rural subjects. Increased age was shown to be an important risk factor for all these disorders, whereas BMI-associated risk was significant with NIDDM and hypertension but not with IGT. PMID- 7497871 TI - Early involvement of central nervous system in type I diabetic patients. AB - OBJECTIVE: To investigate whether the asymptomatic involvement of the central and peripheral nervous systems may be an early complication of diabetes. RESEARCH DESIGN AND METHODS: We studied early impairment of the central and peripheral nervous system pathways in 15 type I diabetic patients with good metabolic control and short disease duration and in 10 healthy control subjects using a set of neurophysiological tests. RESULTS: Results in diabetic subjects showed 1) impairment of motor (7% and somatosensory (13%) pathways of the central nervous system, 2) impairment of motor and sensory conduction velocities (40-60%), and 3) normal values of the vibration perception threshold and cardiovascular autonomic tests. CONCLUSIONS: The damage is more evident in peripheral sites where hyperglycemia and aldose reductase pathways are more active. Instead, several episodes of hypoglycemia, which occur in type I diabetic patients in good metabolic control, may cause alterations of brain nervous cells. PMID- 7497872 TI - Hypoglycemia following inadvertent and factitious sulfonylurea overdosages. AB - OBJECTIVE: To recognize unreported sulfonylurea overdosages in hypoglycemic patients. CASES: We describe three patients with hypoglycemia due to inadvertent (in two patients) and factitious (in one patient) sulfonylurea overdosages. We review the world literature and summarize 43 previously published cases of inadvertently administered and 23 previously published cases of factitiously self administered sulfonylurea overdosages with hypoglycemia. RESULTS: An inadvertently administered fulsonylurea overdosage usually occurred when a sulfonylurea was accidentally substituted for an intended medication with a similar generic or trade name. Features of the patients with a factitiously self administered sulfonylurea overdosage included: 1) a history of the patient or patient's spouse having a medical job or sulfonylurea-treated diabetes mellitus; 2) an unusual affect or psychiatric history; 3) an abrupt onset of severe symptoms without previous milder symptoms; and 4) an absent hypoglycemic or hyperinsulinemic response to provocative testing. These features are not typical for an insulinoma. CONCLUSIONS: When a hypoglycemic patient denies antidiabetic medication use, we recommend sequentially performing: 1) a thorough pill inspection; 2) an interview for recently altered pill appearances; 3) a measurement of serum insulin and C-peptide levels during hypoglycemia; and 4) a blood or urine sulfonylurea screen. Discovery of an unreported sulfonylurea overdosage can eliminate the need to search for an insulinoma and prevent further overdosages from occurring. PMID- 7497868 TI - Decreased erythrocyte membrane fluidity in poorly controlled IDDM. Influence of ketone bodies. AB - OBJECTIVE: To examine the factors that might alter the fluidity of erythrocyte membrane in insulin-dependent diabetes mellitus (IDDM) patients. RESEARCH DESIGN AND METHODS: The subjects were 10 health men and 30 IDDM mem: 10 with good blood glucose (BG) control (HbA1c 5.88 +/- 0.60% [mean +/- SD]), 10 with poor BG control (HbA1C 9.48 +/- 1.05%), and 10 with poor BG control and mild to moderate diabetic ketoacidosis (DKA) (HbA1C 9.12 +/-2.25%, strongly positive ketonuria 3+ and elevated plasma beta-hydroxybutyrate). Erythrocyte membrane fluidity was determined by fluorescence polarization using 6-(9-anthroyloxy stearic acid as fluorescent probe. RESULTS: Membrane fluidity was normal in the diabetic patients with good BG control but significantly lower in the two groups of patients with poor BG control than in the healthy subjects (P < 0.01). The membrane fluidity in the poor BG control groups was also lower in the patients with DKA than in those without DKA (P < 0.01). CONCLUSIONS: The factors that most influence membrane fluidity in IDDM patients appear to be hyperglycemia and ketone bodies. PMID- 7497873 TI - Alterations in the patterns of insulin secretion before and after diagnosis of IDDM. AB - OBJECTIVE: To study the natural history of beta-cell dysfunction in an individual who developed insulin-dependent diabetes mellitus (IDDM) over a 13-month period while under observation. RESEARCH DESIGN AND METHODS: Insulin secretion rates (ISR) in response to intravenous glucose and mixed meals were estimated by deconvolution of C-peptide levels. RESULTS: When fasting glucose and glycosylated hemoglobin concentrations were still within the normal range, insulin secretory responses to intravenous glucose infusion were reduced, but 80- to 100-min secretory oscillations could still be detected. Sequential glucose infusion studies over a 3-month period demonstrated a progressive reduction in insulin secretion. The tight temporal coupling between ultradian oscillations in ISR and glucose observed in nondiabetic subjects was lost. In response to mixed meals, the oscillatory pattern of secretion was preserved, but the magnitude of the secretory responses was reduced. CONCLUSIONS: Our results indicate that despite the lower absolute secretory rates, ultradian ISR oscillations persist in the period before and immediately after the onset of IDDM in this subject, but they are less tightly coupled to glucose than in nondiabetic subjects. PMID- 7497874 TI - Microalbuminuria and potential confounders. A review and some observations on variability of urinary albumin excretion. PMID- 7497875 TI - Absence of shared HLA class II (DR, DQ)-linked genetic basis between IDDM and autoimmune thyroid disease in Japanese. PMID- 7497876 TI - DIDMOAD or Wolfram syndrome. A mitochondrial-mediated disorder. PMID- 7497877 TI - Guar gum in the treatment of NIDDM. PMID- 7497878 TI - Response to Tran et al. PMID- 7497879 TI - The American Diabetes Association Risk Test for diabetes. PMID- 7497880 TI - Islet cell and thyroid autoimmunity in Chinese patients with IDDM. PMID- 7497882 TI - Insights into IDDM at the 1994 EASD conference. European Association for the Study of Diabetes. PMID- 7497881 TI - Accuracy of devices for self-monitoring of blood glucose including hypoglycemic blood glucose levels. PMID- 7497883 TI - Hospice and palliative care. AB - The emergence of AIDS and the aging of the population, with the numerous malignant and debilitating maladies associated with growing older, have focused attention on the provision of cost-effective quality care by hospice and palliative care programs. Hospice and palliative care is a venerated system of care, which uses an interdisciplinary approach to address the medical, psychosocial, and spiritual issues that arise in the treatment of terminally ill patients. This interdisciplinary stratagem for symptom control is necessary to ensure that dying patients and their families are afforded dignity and quality of life through death and the period of familial bereavement. Although death is dominant in palliative situations, terminal care requires an affirmation of life and a recognition that dying is not an aberration of medical care but a natural and normal process. Palliative care, however, also requires a personal acceptance of death and an acknowledgment that dying does not denote a failure to provide good medical care but, rather, calls for an acquiescence that curative treatment is no longer feasible. Accordingly, the terminal state is an integral process and a time to reconcile differences so that patient and family may accept death with a minimum of physical, spiritual, and psychosocial anguish. This article discusses the various precepts cardinal to hospice and palliative care, including the philosophy of terminal care, the management of pain, the adverse effects of analgesic medications, the management of nonpain symptoms, the use of terminal sedation, and the stages of familial bereavement. PMID- 7497885 TI - Correlation between Goldmann perimetry and maximal electroretinogram response in retinitis pigmentosa. AB - To evaluate the relationship between Goldmann perimetry and maximal electroretinographic responses in patients with retinitis pigmentosa, analyses were performed on 220 affected subjects and separately on two subgroups with autosomal dominant (n = 35) and autosomal recessive (n = 29) inheritance. Electroretinograms were recorded averaging 100 iterations elicited with a 20 lux/s, 0.5-Hz white flash ganzfeld stimulation. The peripheral isopters of the visual fields were delimited with I4e, IIIe and V4e targets, measured on conventional perimetry charts with a light pen and expressed in square centimeters. Unlike most previously published reports, this investigation showed a definite correlation (p = 0.0001) between maximal electroretinographic response amplitude and visual field areas. This correlation was more evident for I4e and IIIe isopters (r = 0.89 and 0.87, respectively) than for V4e isopter (r = 0.69). This phenomenon appears to be related to distortion occurring on standard isometric charts and to spatial summation effects in the peripheral field. Such correlations held for both the autosomal dominant and autosomal recessive subgroups. It appears that, if enough accuracy is provided, maximal electroretinographic responses and Goldmann visual fields are both good measures of the remaining functioning retina in nonsyndromic retinitis pigmentosa, irrespective of inheritance models and dystrophic patterns. PMID- 7497884 TI - Postnatal retinal development as measured by the electroretinogram in premature infants. AB - Light-adapted and dark-adapted electroretinograms were obtained in 27 premature infants who were screened for retinopathy of prematurity shortly after birth. Thirteen showed no retinopathy and 13 had mild to moderate retinopathy, which ranged from stage I, zone III (International Classification) to stage III, zone II. Measurements were made during the first 16 months of life. The configuration of the waveforms under both photopic and scotopic conditions changed during this period showing increased amplitudes of both the a- and the b-waves. A scotopic intensity series was performed and the b-wave amplitudes and implicit times were measured. The b-wave amplitude data were averaged for three adult control subjects, for infants without retinopathy of prematurity measured at 3-4 and at 6 7 months and for infants with retinopathy of prematurity measured at 3-4 and at 6 7 months. The Naka-Rushton function was fitted to the average data. The Rmax increased from 3 to 6 months and from 6 months to adulthood, and the Isat values decreased over this age range. The b-wave implicit times were within normal range in the 6-month data. There was no difference in Rmax or Isat for infants with and without ROP. PMID- 7497886 TI - Surgical therapy for obesity can induce a vitamin A deficiency syndrome. AB - New possible causes and unexplored aspects of the electroretinogram were evaluated in a case of vitamin A deficiency secondary to surgical therapy for morbid obesity. The Naka-Rushton equation, applied to the scotopic b-wave, demonstrated the quantal catch reduction caused by the loss of rhodopsin in the outer segment of photoreceptors. Study of the Fourier analysis of the photopic 20 Hz response suggested a primary involvement of the external retinal layers, with an indirect alteration of the inner layer. The electroretinogram oscillatory potentials showed alterations explained by the involvement of their generators related to the primary photoreceptor lesion. PMID- 7497887 TI - The first and second harmonics of the macular flicker electroretinogram: differential effects of retinal diseases. AB - We evaluated the effects of retinal diseases on the macular electroretinogram first and second harmonic components, which are dominated by outer and inner retinal activity, respectively. Macular electroretinograms in response to a uniform field (9 degrees x 9 degrees) flickering sinusoidally at either 32 or 8 Hz (peak frequencies of the first and second harmonics, respectively) were recorded in 14 patients with maculopathies involving photoreceptors (e.g., age related macular degeneration), in 16 patients with postreceptoral macular diseases (e.g., branch occlusion of central retinal artery), and in 38 normal controls. Amplitude and phase of the first and second harmonic response components were evaluated by Fourier analysis. When compared to controls, patients with photoreceptor diseases had reduction in both first and second harmonic mean amplitudes and second harmonic phase delay; patients with postreceptoral diseases had normal first harmonic components but reduced and delayed second harmonic components. A discriminant analysis, by using first and second harmonic values, correctly classified 13 of 14 patients with photoreceptor diseases and 14 of 16 patients with postreceptoral disorders. These results indicate that combined evaluation of the macular electroretinogram first and second harmonic components is a useful test for identifying the site(s) of retinal dysfunction in patients with macular diseases. PMID- 7497888 TI - Spatial-temporal interactions in the steady-state pattern electroretinogram. AB - It is currently assumed that steady-state pattern electroretinograms can be obtained only at high rates of pattern reversal. However, steady-state responses can also be obtained at very low temporal frequencies (less than 1 Hz), provided that the reversal is sinusoidal. In five healthy volunteers, we studied the frequency characteristics of the pattern-reversal electroretinogram in response to sinusoidal gratings of 0.4 and 4 c/deg, reversed sinusoidally in contrast at frequencies ranging from 1 to 27 Hz. Steady-state responses dominated by the second harmonic component were obtained at all temporal frequencies tested; the amplitude of the second harmonic changed with stimulus temporal frequency. In the low-temporal-frequency range, stimuli of high spatial frequency appear to elicit larger contribution of generators with sustained or tonic characteristics compared with stimuli of low spatial frequency. PMID- 7497889 TI - Long wavelength-middle wavelength cone interaction under no background in the electroretinogram of the cynomolgus monkey. AB - We recorded electroretinograms from anesthetized cynomolgus monkeys for 30 monochromatic (400-700 nm) full-field stimuli with five different stimulation levels presented without a background light. Waveforms were analyzed by means of principal component analysis to investigate the influence of stimulus intensity on long- and middle-wavelength cone interaction. With increasing levels of stimulus intensity, the middle-wavelength cone system showed a slight peak shift and a noticeable increase in response, while the long-wavelength cone system showed an obvious change in peak wavelength rather than response. In addition, spectral responses in the long-wavelength cone system increased over the red region, whereas they decreased over the yellow region. On the other hand, responses in the middle wavelength cone system increased over the yellow region much more than over the green region. These complicated changes are thought to indicate the induction of long- and middle-wavelength cone interaction by stimulus light. PMID- 7497891 TI - Visual evoked cortical potentials and psychophysical determination of color contrast thresholds along different chromatic axes. AB - Color vision can be assessed by examining the color contrast threshold along various color axes. We investigated the possibility of determining these thresholds objectively by means of visual evoked cortical potentials. A color calibrated flicker-free (112-Hz) monitor and a 14-bit-per-gun board allowed visualization of colors with specified xyY CIE 1931 coordinates. Horizontal grating, 1 c/deg were sinusoidally alternated at 8 Hz for both visual evoked cortical potential recordings and psychophysical determinations. Two healthy emmetropic 35 year-old subjects performed color brightness matching along each color axis, before any recording and reduction in color contrast. For each color axis, extrapolation to zero voltage of the visual evoked cortical potential amplitude versus log color contrast response allowed determination of the color contrast threshold. The visual evoked cortical potential-derived threshold changed considerably with the color axis, with evident intersubject differences. These differences were similar to those observed in the psychophysically determined thresholds. Visual evoked cortical potential responses to suitable chromatic stimuli allow determination of color contrast thresholds that correspond well to those determined psychophysically. Hence, with the visual evoked cortical potential, accurate objective assessment of color vision is feasible and may be useful in both research and clinical settings. PMID- 7497890 TI - The effects of lidocaine and bupivacaine on the rabbit retina. AB - The toxic action of two commercial anesthetics, lidocaine and bupivacaine, on the functional and morphologic integrity of the retina was investigated in albino and pigmented rabbits. The experimental drug was injected into the vitreous of one eye, while saline solution was injected into the fellow eye. Retinal function was assessed from the electroretinogram and the visual evoked potential. Retinal structure was examined at the light microscopic level. Ten milligrams of lidocaine did not affect the electroretinogram and the visual evoked potential responses, though structural damage could be detected close to the site of injection. A lower dose of 5 mg did not produce any detectable physiologic or morphologic damage. The only dose of bupivacaine used, 0.5 mg, was not toxic to the albino and pigmented rabbit retinas, as assessed by the electroretinogram, visual evoked potential, and light microscopy. The results of this study demonstrate that lidocaine and bupivacaine are nontoxic to the rabbit retina at concentrations that are effective for retrobulbar anesthesia. PMID- 7497892 TI - [Endoscopic retroperitoneal adrenalectomy]. AB - AIM OF STUDY: To determine prospectively in consecutive patients the value of endoscopic retroperitoneal adrenalectomy. PATIENTS AND METHODS: Between March 1994 and March 1995, endoscopic adrenalectomy via a retroperitoneal approach was performed in eleven patients (three men, eight women; median age 61 [48-73] years), unilateral in nine, bilateral in two. The procedure was indicated if the adrenal tumour was thought to be benign and no larger than 5 cm in diameter (two adrenal and two central Cushing's syndromes; three incidentally detected adrenal adenomas, two phaeochromocytomas, two Conn adenomas). RESULTS: Twelve of the 13 procedures were successfully performed. In one case the primary incision in the mid-axillary line had been too large and no satisfactory pneumoretroperitoneum was achieved. Median duration of operation was 180 (125-330) min, and the intraoperative blood loss was 200 (125-330) ml. There were no postoperative complications and the median postoperative hospital stay was 6 days (3-12). CONCLUSION: In small (up to 5 cm) adrenal tumours not suspected to be malignant the endoscopic retroperitoneal approach constitutes a sparing alternative to the conventional access to the adrenal gland. PMID- 7497893 TI - [Rheumatoid arthritis, neutropenia and splenomegaly: the Felty syndrome]. AB - HISTORY AND FINDINGS: A 59-year-old asymptomatic man, first diagnosed to have rheumatoid arthritis 27 years ago, was admitted to hospital because of splenomegaly and neutropenia, first noted 2 years ago. Physical examination confirmed splenomegaly and also revealed pretibial hyperpigmentation, but no evidence of active rheumatoid arthritis. EXAMINATIONS: Biochemical tests showed relative and absolute neutropenia (white blood cell count 2200/microliters; 1% neutrophils), thrombocytopenia and polyclonal hypergammaglobulinaemia. He also had increased erythrocyte sedimentation rate (38/92), a high titre of rheumatic factor (2128 IU/ml) and increased circulating immune-complexes (74%). Thoracic and abdominal computed tomography provided no evidence of malignant tumor. The spleen measured 15 x 7 x 10 cm. Bone-marrow biopsy from the iliac crest revealed abnormal maturation of granulopoiesis and marked lymphoid infiltration. The clinical triad of rheumatoid arthritis, splenomegaly and neutropenia are diagnostic of Felty's syndrome. As the patient was asymptomatic there was no indication for treatment. CONCLUSION: Felty's syndrome is a rare condition demanding considerable effort in differential diagnosis. PMID- 7497895 TI - [Hepatic echinococcosis with gallstones of the echinococcal cavity]. AB - HISTORY AND CLINICAL FINDINGS: A 65-year-old woman was hospitalized with suspected cholelithiasis. The only contributory item in the history was jaundice of uncertain cause 15 years previously. Physical examination elicited pain in the right upper abdomen on deep palpation. INVESTIGATIONS: Alkaline phosphatase and gamma-GT activities were raised (324 and 407 U/ml, respectively). Ultrasound revealed cholecystolithiasis; the bile duct was 5 mm in diameter. Intravenous contrast-medium cholangiography showed choledocholithiasis and circular flat calcification in the bifurcation of the hepatic duct. Computed tomography revealed this structure to be a space-occupying mass (10 x 6 x 5 cm), with a calcified border (density of 27 Hounsfield units), raising the suspicion of a blood-containing hydatid cyst. Additionally there were two calculi in the left hepatic duct. But the echinococcus test (by indirect haemagglutination and enzyme linked immunosorbent assay) was negative. TREATMENT AND COURSE: Endoscopic retrograde cholangiography (ERC) with papillotomy was performed and two choledochal concrements removed. Concrements within the mass were also visualized. Obstructive jaundice developed 3 days after the ERC and a laparotomy was performed. Excision of the hydatid was not possible because the stone-filled hydatid cavity could not be punctured. Choledochal exploration discovered membranes which histologically were chitin-like structures and corresponded to scolices. Under chemical litholysis the hydatid became smaller. In addition, albendazole was given in two four-week cycles (400 mg twice daily). The patient quickly recovered and 5 months later was symptom-free. PMID- 7497894 TI - [An anticholinergic syndrome with hallucinatory psychosis after diphenhydramine poisoning]. AB - HISTORY AND CLINICAL FINDINGS: A 27-year-old female student was hospitalized as an emergency because of gait and speech disorders which had developed over just a few hours. Conflicts with her partner and frequent alcohol abuse were reported. It was not possible to interview the patient herself. Consciousness was clouded by optical and acoustic hallucination. Body temperature was 38.1 degrees C: the skin was warm and dry, the tongue dry. Heart rate was 120/min, blood pressure 150/100 mm Hg. Neck and limb muscle tone was increased. Hallucinatory psychosis, encephalitis and alcohol withdrawal delirium were considered in the differential diagnosis. TESTS: There was hypokalaemia, slightly increased serum creatinine and metabolic alkalosis. The alcohol level was 0.18%. The ECG was normal except for sinus tachycardia. TREATMENT AND COURSE: In the meantime, two empty packets of 20 tablets each had been found in the patient's flat. They had contained Vivinox and Sediat (total of 1.5 g diphenhydramine), so that a central anticholinergic syndrome due to an overdose with this drug could be assumed. The diphenhydramine level was 2.8 micrograms/ml. After administration of altogether four times 2 mg physostigmine and 5 mg diazepam intravenously and normalization of electrolytes the symptoms regressed within 12 hours. The patient then admitted to having taken the drugs with suicidal intent. She was transferred to the psychiatric department because of the risk of further suicidal attempts. PMID- 7497896 TI - [The therapy of viral hepatitis in HIV-infected patients]. PMID- 7497897 TI - [Chronic aortic valve insufficiency. The current status of its diagnosis and therapy]. PMID- 7497899 TI - [Brain functional disorders after taking Ecstasy?]. PMID- 7497900 TI - [ACE inhibitor dosage in heart failure]. PMID- 7497898 TI - [Once more: the duty of fee reduction according to Article 6 a of the Schedule of Charges for Physicians]. PMID- 7497901 TI - [Vitamin D prophylaxis in children]. PMID- 7497903 TI - [Urinary calculus analyses]. PMID- 7497902 TI - [Urinary calculus analyses]. PMID- 7497904 TI - [The prophylactic intensification of the therapy in systemic lupus erythematosus]. PMID- 7497905 TI - [The safety and efficacy of saquinavir in the therapy of HIV infection]. PMID- 7497906 TI - Protective effect of nifedipine against cytotoxicity and intracellular calcium alterations induced by acetaminophen in rat hepatocyte cultures. AB - Alteration of calcium homeostasis has been proposed to play a major role in cell necrosis induced by a variety of chemical agents such as acetaminophen (APAP). In this study, a potential protective effect of the dihydropyridine calcium channel blocking agent, nifedipine, was investigated in vitro on acetaminophen-induced hepatocyte damage. Rat hepatocytes were exposed during 20 hours to various concentrations of APAP (0.50 to 4.00 mM). The following metabolic and functional parameters were investigated: -lactate dehydrogenase (LDH) release as an indicator of plasma membrane integrity, -cell viability evaluated by the colorimetric MTT assay, and intracellular calcium concentration as evaluated by two fluorimetric methods: a scanning laser cytometer using indo-1-AM as fluorescent probe and a fluorescence plate reader using fluo-3-AM as calcium indicator. Incubation of hepatocytes with APAP alone in the range 0.50 to 4.00mM resulted in a dose-response relationship with regard to LDH release (243% to 750% of control) and to the loss of cell viability (0 to 67% of control). Moreover these results were correlated with a significant increase in cytosolic calcium content (189 to 406 nM). Nifedipine treatment prior to APAP exposure, partially prevented LDH release, the plasma membrane blebbing, and thereby the loss of viability. In addition, intracellular calcium level progressively returned within the limits of the control values with increasing concentrations of nifedipine. It can be concluded that, in vitro conditions, nifedipine pretreatment exhibits a preventive effect against acetaminophen hepatocyte injury. PMID- 7497908 TI - Effects of atropine and soman on the pharmacokinetics of the oxime HLo 7 dimethanesulfonate in anesthetized guinea-pigs. AB - The respiratory and circulatory effects and the pharmacokinetics of the bispyridinium dioxime HLo 7 (30 mumol/kg) were investigated in guinea-pigs in combination with atropine (28.8 mumol/kg) or with atropine plus soman (0.44 mumol/kg = 5 x LD50). The pharmacokinetics and pharmacodynamics were studied in separate groups of animals. HLo 7 had only minor effects on circulation and respiration. HLo 7 plus atropine caused a transient, pronounced fall of blood pressure, tachycardia, and respiratory stimulation. The soman-induced bradycardia was completely restored by the antidotes, while respiration improved within a few minutes with the respiratory rate becoming stabilized at 50% of baseline. Plasma half-time (60-70 min), plasma clearance (6-7 ml.kg-1.min-1) and apparent volume of distribution (0.5-0.6 l/kg) of HLo 7 did not differ between the groups, i.e. atropine and soman did not affect the kinetics of the oxime. PMID- 7497907 TI - Dose-response effects of atropine and HI-6 treatment of organophosphorus poisoning in guinea pigs. AB - HI-6 (1-2-hydroxyiminomethyl-1-pyridino-3-(4-carbamoyl-1-pyridino -2- oxapropane dichloride) has been evaluated as an oxime alternative to pralidoxime, and toxogonin in the treatment of organophosphorus (OP) poisoning. The dose response effects of atropine (ATR) and HI-6 were investigated to more fully explore the interaction of these compounds in the treatment of OP poisoning. ATR, HI-6 and various combinations of the two drugs were evaluated against lethal poisoning by soman (GD) and tabun (GA) in guinea pigs. The effect of adjunctive diazepam treatment on the efficacy of atropine and HI-6 against soman was also investigated. Animals of either sex were challenged s.c. with OP and treated i.m. 1 min later with ATR and/or HI-6. When used, diazepam was injected immediately after ATR+HI6. LD50s of each treatment were calculated from probit models based on 24-hour survival against 5 levels of nerve agent and 6 animals per challenge level. A protective index (PI) was calculated by dividing the nerve agent LD50 in the presence of treatment by the LD50 in the absence of treatment. Treatment with HI6 alone had little effect on the toxicity of either OP. Treatment with ATR alone was more effective than HI-6 alone and was significantly more effective against soman than against tabun. When used in combination atropine and HI-6 had a strong synergistic effect against both agents. The dose of atropine used with HI-6 was critical in determining the efficacy of HI-6 against either agent. The slopes of the dose-lethality curves were minimally affected by the dose of ATR or HI-6. Adjunctive treatment with diazepam enhanced the efficacy of HI-6 and atropine against soman. It is concluded that 1) ATR has a large effect on the efficacy of HI-6 against OP poisoning, 2) the dose of ATR must be carefully selected in studies investigating the efficacy of HI-6 against OP poisoning, 3) the effective dose of ATR in the guinea pig is approximately 16 mg/kg, and 4) diazepam is a useful adjunct to atropine and HI-6. PMID- 7497909 TI - Diethyldithiocarbamate protection against cisplatin nephrotoxicity: antioxidant system. AB - This investigation elucidates the role of antioxidant system in cisplatin induced nephrotoxicity and the nephroprotection with diethyldithiocarbamate (DDTC). Male Wistar rats were injected with 1) cisplatin; 2) cisplatin+DDTC and 3) vehicle control. Rats were sacrificed three days post-treatment and the corticomedullary junction of the kidney was isolated adn were analyzed for GSH, GSSG, SOD, CAT, and GSH.Px. Serum creatinine increased (500% of control) following cisplatin administration which decreased to 200% of control with DDTC. Cisplatin treated rats showed depletion of GSH levels, while cisplatin+DDTC injected rats had GSH values similar to controls. SOD and GSH.Px activities were found to be 63 and 40% of control following cisplatin administration which increased to 109 and 75% of control with DDTC respectively. Our findings suggest that cisplatin nephrotoxicity is mediated by impaired activities of SOD and GSH-Px enzymes and by GSH depletion. The protective mechanism of DDTC against cisplatin nephrotoxicity is related to the prevention of GSH depletion and restoring SOD and GSH-Px activities in the kidney of rats. PMID- 7497910 TI - The effects of subacute and subchronic oral exposure to cis-1,2-dichloroethylene in Sprague-Dawley rats. AB - Cis-1,2-dichloroethylene was administered daily by corn oil gavage to male and female Sprague-Dawley rats at the following dose levels: 1.0, 3.0, 10.0 and 22.0 mmol/kg/day for 14 days. Doses gavaged during the 90-day subchronic study were 0.33, 1.00, 3.00 and 9.00 mmol/kg/day. There were no compound-related deaths or histopathological changes demonstrated. Significant increases in relative liver weights were seen after 14- and 90-days of treatment in both sexes. This study demonstrates some indication of toxicity at subacute and subchronic exposure levels as low as 0.33 mmol/kg/day. Implications of liver abnormalities were demonstrated at an exposure level of 1 mmol/kg/day while kidney abnormalities (relative weights) were demonstrated at an exposure level of 0.33 mmol/kg/day. PMID- 7497911 TI - The effects of subchronic chlorate exposure in Sprague-Dawley rats. AB - Male and female Sprague-Dawley rats were exposed to drinking water containing 3.0, 12.0 or 48.0 mM sodium chlorate. The mean drinking water consumption varied between exposure groups from 100-200 ml/kg/day. Female exposure groups consistently drank more water (23-42%) than male exposure groups thereby receiving more chlorate/kg/day at every exposure level. There were no compound related deaths; however, both males and females in the high exposure groups had significant weight loss during the 90-day exposure period. Also, in these same groups females had mild but significant decreases in the following relative organ weights; adrenals, thymus and spleen, while the relative brain weight was increased. In males, the heart, kidneys and liver were mildly decreased while the brain and testes were mildly increased. Red blood cell counts and percent hematocrit were decreased in both sexes in the high dose group. Pituitary gland (pars distalis) vacuolization and thyroid gland colloid depletion were prominent in both sexes in mid and/or high dose animals. A NOAEL of 0.36 mM chlorate/kg b.w./day in males and 0.50 mM chlorate/kg b.w./day in females were established. PMID- 7497914 TI - [SETTI-Study and antioxidants--more questions than answers]. PMID- 7497913 TI - Haematological changes in the garden lizard, Calotes nemoricola due to the defensive secretion of the grasshopper, Poecilocerus pictus. AB - Defensive secretion of the Grasshopper, Poecilocerus pictus produced significant changes in the blood components of the predator lizard, C. nemoricola. The red blood cell (RBC) count, haemoglobin (Hb) concentration, packed cell volume (PCV), mean corpuscular haemoglobin (MCH), mean corpuscular haemoglobin concentration (MCHC) and white blood cell (WBC) count decreased greatly, whereas mean corpuscular volume (MCV) showed significant increase following the administration of lethal dose of defensive secretion. These changes however were found to be reversible toward control levels in the lizards injected with sublethal dose. Disrupted erythropoeitic activity in lizards by the cardenolides in the defensive secretion was suggested. PMID- 7497912 TI - Toxicity studies of 1,3-dichlorobenzene in Sprague-Dawley rats. AB - Male and female Sprague-Dawley rats received 1,3-dichlorobenzene daily by corn oil gavage for 10 or 90 consecutive days. The 10-day study doses were 0, 37, 147, 368 and 735 mg/kg; the 90-day study doses were 0, 9, 37, 147 and 588 mg/kg. In the 10-day study, there was a significant depression of body weight in both sexes at 735 mg/kg. Liver weights were significantly increased in both sexes at 368 and 735 mg/kg. Serum cholesterol levels were significantly elevated in both sexes at 368 and 735 mg/kg. Histopathological evaluation revealed centrolobular hepatocellular degeneration at 368 mg/kg in males and 735 mg/kg in females. In the 90-day study, body weights were significantly depressed in both sexes at 588 mg/kg. Normalization of food and water consumption by final body weight indicated that at 588 mg/kg both sexes had increased food and water consumption relative to controls. Absolute and relative liver weights were significantly increased in both sexes at 147 and 588 mg/kg. Relative kidney weights were significantly higher in both sexes at 588 mg/kg and in males at 147 mg/kg. Serum cholesterol and calcium levels were significantly elevated over controls in females at 37, 147, and 588 mg/kg, and in males at all dose levels. Histopathological evaluation at 147 and/or 588 mg/kg demonstrated liver and thyroid lesions in both sexes, and pituitary and kidney lesions in males. A NOAEL was not firmly established. PMID- 7497916 TI - [Autologous stem cell support allows use of high-dose chemotherapy in breast cancer]. PMID- 7497915 TI - [Raised blood glucose level increases risk of atherosclerosis in non-insulin dependent diabetes]. PMID- 7497917 TI - [A girl or a boy, as made to order]. PMID- 7497918 TI - [TOS (thoracic outlet syndrome)--a functional disease?]. PMID- 7497919 TI - [DEPS--a new tool in screening for depression]. PMID- 7497920 TI - [A victim of lightning strike]. PMID- 7497921 TI - [Dissection of a vertebral artery as a complication of neck manipulation]. PMID- 7497923 TI - [Variable results on nuclear hormone receptors]. PMID- 7497922 TI - [Treatment of seborrhoeic dermatitis]. PMID- 7497924 TI - [Exercise, health and health services]. PMID- 7497925 TI - [Does professional sports need psychiatry?]. PMID- 7497926 TI - [Contraindications for physical training]. PMID- 7497927 TI - [Physical activity and health: minimum, optimum and maximum]. PMID- 7497928 TI - [Physical activity and prevention and treatment of metabolic syndrome]. PMID- 7497929 TI - [Physical activity and musculoskeletal functioning]. PMID- 7497931 TI - [Can virus safety of blood products be assured?]. PMID- 7497932 TI - [Should a brain-injury patient be rehabilitated?]. PMID- 7497933 TI - [Paclitaxel, a new anticancer drug]. PMID- 7497930 TI - [Drug treatment and exercise]. PMID- 7497934 TI - [Brain neurochemical systems which control appetite]. PMID- 7497935 TI - [Chronic hepatitis among Finnish patients with hemophilia]. PMID- 7497936 TI - [Free living amebas in whirlpools and swimming pools--a risk for contact lens users]. PMID- 7497938 TI - [Nocturnal bruxism with severe facial and neck pain]. PMID- 7497937 TI - [Cholesterol embolism--a rarely diagnosed common phenomenon]. PMID- 7497939 TI - [Dopa-responsive dystonia]. PMID- 7497940 TI - [Diet in celiac disease]. PMID- 7497945 TI - [Apolipoprotein E polymorphism and serum cholesterol levels in Finnish children and adolescents]. PMID- 7497941 TI - [Jorma Palo and euthanasia]. PMID- 7497944 TI - [Population biology of death]. PMID- 7497942 TI - ['Salt' hypertension also among women]. PMID- 7497943 TI - [General anticoagulant factor deficiency related to venous thrombosis has been identified]. PMID- 7497946 TI - [Side effects of phenytoin and carbamazepine on cognitive functioning]. PMID- 7497947 TI - [Iron prophylaxis and maternal hematological levels during pregnancy]. PMID- 7497948 TI - [Pustules in finger tips--antibiotics, amputation, anamnesis]. PMID- 7497949 TI - [Marchiafava-Bignami disease--an underdiagnosed syndrome?]. PMID- 7497950 TI - [For safety's sake]. PMID- 7497951 TI - [Herniated thoracic disc]. PMID- 7497952 TI - [Treatment of intestinal parasitic diseases]. PMID- 7497953 TI - [Bank robbery and psychiatric insurance]. PMID- 7497955 TI - [Physician's role as a rehabilitation worker]. PMID- 7497954 TI - [Was the dosage correct?]. PMID- 7497956 TI - [Goals in rehabilitation]. PMID- 7497957 TI - [Financing of rehabilitation and administrative reform]. PMID- 7497958 TI - [Responsibility to arrange treatment and rehabilitation services, financing and accessibility of services from the viewpoint of the Social Insurance Institute]. PMID- 7497959 TI - [Evaluation and development of rehabilitation legislation]. PMID- 7497960 TI - [Rehabilitation as part of municipal health care center's functions]. PMID- 7497962 TI - [Rehabilitation of cardiac patients]. PMID- 7497961 TI - [The whole picture of rehabilitation]. PMID- 7497963 TI - [Adolescents and rehabilitation]. PMID- 7497964 TI - [Stroke rehabilitation]. PMID- 7497965 TI - [How to facilitate the rehabilitation of patients with musculoskeletal symptoms]. PMID- 7497966 TI - [Rehabilitation of patients with respiratory system diseases]. PMID- 7497967 TI - [Rehabilitation of cancer patients]. PMID- 7497968 TI - [A rehabilitation project for patients with schizophrenia]. PMID- 7497969 TI - [Methods in the economic assessment of rehabilitation]. PMID- 7497970 TI - [Doctor-patient relationship in rehabilitation]. PMID- 7497971 TI - [Current trends in vocational rehabilitation]. PMID- 7497972 TI - [Environmental control systems for severely handicapped persons]. PMID- 7497973 TI - [Hope for amyotrophic lateral sclerosis?]. PMID- 7497974 TI - [Brakes of the cell cycle--new inhibitor genes?]. PMID- 7497975 TI - [Cupping as evaluated by the client]. PMID- 7497976 TI - [Threats and physical violence at work]. PMID- 7497977 TI - [Recurrent brief depression]. PMID- 7497978 TI - [The patient-doctor relationship and metabolic control in patients with diabetes mellitus]. PMID- 7497980 TI - [Treatment of prolonged head-and-neck pain by balancing occlusional problems]. PMID- 7497979 TI - [Foreign body in the bladder as a cause of lower abdominal pain and recurrent urinary tract infections in a female patient]. PMID- 7497981 TI - [Streptokinase treatment for thrombosis of a tricuspid valve prosthesis]. PMID- 7497983 TI - [Problem-oriented education, problem-based learning and activating teaching methods]. PMID- 7497982 TI - [Magnetic resonance or computerized tomography for imaging of the brain?]. PMID- 7497984 TI - [From dinosaurs to dialogue]. PMID- 7497985 TI - [Untreated teeth--a health risk]. PMID- 7497986 TI - [Immunological tolerance in prevention of diabetes]. PMID- 7497987 TI - [Cytochrome P-450 isoenzymes as predictors of drug interactions]. PMID- 7497988 TI - [Questionnaires as research tools--Social Insurance Institution's Sleep Habit Questionnaire as an example]. PMID- 7497990 TI - [Progressive lung fibrosis following farmer's lung disease]. PMID- 7497989 TI - [Swollen breast in a female patient with dermatomyositis]. PMID- 7497991 TI - [Free radicals and antioxidants. Advantages of antioxidants in clinical medicine, truth or fiction?]. PMID- 7497992 TI - [Why and how the results of antioxidant studies are reported]. PMID- 7497993 TI - [Role of free radicals and antioxidants in the cell defense system]. PMID- 7497994 TI - [Role of vitamin E and C, carotenoids, ubiquinone and selenium in the antioxidative defense system]. PMID- 7497995 TI - [Safety of antioxidants]. PMID- 7497996 TI - [Role of lipid oxidation in the development of atherosclerosis]. PMID- 7497997 TI - [Oxidative stress and cell aging]. PMID- 7497998 TI - Blood transfusion. PMID- 7497999 TI - Surgical management of patients with severe anaemia due to acute blood loss: a case for withholding perioperative blood transfusion. AB - Twenty one consecutive patients who had an average packed cell volume (PCV) of 16.2% (range 13-25) due to acute blood loss were treated surgically without perioperative blood transfusion. All the surgical procedures were carried out under intravenous ketamine hydrochloride. The surgical wound in each patient healed by primary intention. The packed cell volume rose to 35% and above in an average of six weeks by use of oral haematinics. The only morbidity observed was dizziness which lasted for an average of six days in ten patients. There were no deaths and all the patients remained well at an average follow-up period of 42 weeks. The study shows that perioperative blood transfusion can be avoided in surgical care of most patients who have severe anaemia due to acute blood loss without mortality and without significant morbidity. PMID- 7498000 TI - Attitudes and beliefs about blood donation among adults in Mwanza Region, Tanzania. AB - A study was carried out to assess the attitudes and beliefs about blood donation among the population of Mwanza Region, Tanzania, in order to target better the voluntary blood donor recruitment and retention. A random sample of 1141 adults, 743 residents of Mwanza town and 398 from Mwanza rural areas were interviewed on aspects related to blood donation. A high proportion (26.4%) of interviewees had already given blood but only 3.8% had donated voluntarily within the last 10 years. Voluntary blood donation was correlated with secondary school attendance. We noted a positive attitude towards voluntary blood donation although the majority of people would do so only for an incentive in the form of remuneration. Respondents frequently thought that blood donation would infect them with HIV or damage their health. No correlation was found between fear to donate and self perceived risk for HIV infection. We found that a substantial number of men would accept pre-test counselling on HIV in order to "infect others" if found positive. Thus we recommend that HIV counselling should be done with extreme care and defer individuals with wrong intentions from blood donation. Donor recruitment campaigns should also focus on clearing wrong conceptions about blood donation through providing information on all aspects related to blood donation. Based on these findings, a scheme for blood donor recruitment and retention in Mwanza Region is proposed. PMID- 7498002 TI - Malignant lymphoma in western province of Saudi Arabia. AB - A review of all malignant lymphomas seen at the King Khalid National Guard Hospital, Jeddah, Saudi Arabia between January 1985 and June 1993 was done. There were 106 cases of non-Hodgkin's lymphoma with a male: female ratio of 2.1:1. Low grade, intermediate grade and high grade lymphomas constituted 17.9%, 36.8% and 28.7% respectively. A comparatively lower frequency of follicular lymphomas (5.6%) was noted, compared to the Western literature. Of the few cases immunophenotyped, T-cell and B-cell tumours make up 29% and 71%, respectively. PMID- 7498001 TI - Response of Plasmodium falciparum to chloroquine and Fansidar in vivo and chloroquine and amodiaquine in vitro in Uganda. AB - The response of P. falciparum to chloroquine and pyrimethamine-sulfadoxine in vivo and chloroquine and amodiaquine in vitro was investigated in parasitaemic school children from six locations. Mean parasite sensitivity to chloroquine at day 7 was 74% (range 61-97) with parasite clearance rates between 2-3 days and complete defervescence in 85% of febrile children. Sensitivity declined in the four sites followed up to day 14 to 45% (range 37-53). Parasites were significantly more sensitive to pyrimethamine/sulfadoxine at 5/6 sites (100% day 7) but 5% of subjects became parasitaemic by day 14. In vitro isolates were significantly less sensitive to chloroquine than to amodiaquine with a mean 99% effective concentration of 348 mumol/L compared to 6.44 mumol/L. Clearly the role of chloroquine as the primary therapy for uncomplicated P. falciparum malaria should be reconsidered especially in the light of increasing disease severity and resurgence. Amodiaquine may be suitable alternative with pyrimethamine/sulfadoxine as second line and for more severe malaria prior to referral. The cost of alternative antimalarials and the dynamic and deteriorating pattern of resistance are powerful arguments for more objective slide diagnosis to minimise drug pressure and a regular drug sensitivity surveillance system. We believe that the latter should concentrate on measuring clinical drug efficacy in symptomatic outpatients rather than in asymptomatic children while the former needs more pragmatic and economical strategies possibly centred on seasonality and risk. PMID- 7498003 TI - Epidemiological and clinical aspects of carcinoma of penis at Kenyatta National Hospital. AB - Thirty one patients with carcinoma of penis were studied retrospectively at Kenyatta National Hospital, Nairobi, over a 20-year period (1971-1990). The majority of patients presented late with symptomatology of over one year duration. 88% of patients with carcinoma were uncircumcised, while the three (12%) patients who were circumcised but developed carcinoma were all circumcised late in adolescence and adulthood, confirming that late circumcision may not protect one from developing penile carcinoma as reported in literature. These findings also indicate that carcinoma of penis may be rare in this locality but is still common among the uncircumcised African tribes. PMID- 7498004 TI - The prevalence of hepatitis C virus antibodies in renal patients, blood donors and patients with chronic liver disease in Kenya. AB - We tested serum samples from four categories of patients with nephrological problems (nephrotic syndrome, stable chronic renal failure, haemodialysis patients and renal transplant recipients), patients with chronic liver disease and volunteer blood donors for the presence of antibody to hepatitis C virus (HCV). Screening was done by second-generation enzyme linked immunosorbent assay (ELISA) and confirmation with second-generation recombinant immunoblot assay (RIBA). Of all the renal patients, only 6.3% of the transplant patients tested positive for anti-HCV, while in patients with chronic liver disease anti-HCV was detected in 2.6% of the patients with chronic hepatitis and in none with liver cirrhosis or hepatocellular carcinoma. This finding of low prevalence in these patient groups was not in keeping with findings in studies done elsewhere. Our anti-HCV prevalence of 0.9% in blood donors was comparable to that found in Europe, USA and Taiwan. We recommend that the low prevalence of anti-HCV in some of our high risk groups should not lead to complacence and hence further studies are necessary to evaluate the infectivity of anti-HCV positive patients and the potential for cross infection. PMID- 7498005 TI - Decline of Glossina morsitans ugadensis in Gambella, Ethiopia. AB - Gambella is the only area where sleeping sickness is endemic in Ethiopia. Four species of Glossina had been reported from Gambella out of the five species found in the country in surveys made before 1985. These are Glossina morsitans ugadensis, G. pallidipes, G. fuscipes and G. tachinoides. A tsetse fly survey was carried out in parts of Gambella owing to the fact that the area is undergoing ecological changes due to massive deforestation (because of resettlement and development programmes), poaching, and introduction of domestic animals into tsetse infested parts of Gambella after 1985. Tsetse populations were sampled for one year, March 1993-April 1994, using biconical traps and hand catches. The survey has reported all Glossina spp which were previously reported except G. morsitans ugadensis. It seems that a combination of factors, such as, lack of host and increase in human population have forced G. morsitans ugadensis to decline. This study has consolidated the fact that tsetse flies of the morsitans group specially G. morsitans, are easily affected by human interference while the palpalis group is resistant to this factor. In addition, this study has also indicated, villagization and rural development could be practised where G. morsitans is the only species in a certain area to alleviate pressure on already impoverished land in parts of Africa. PMID- 7498007 TI - An outbreak of typhoid fever among children who attended a potluck dinner at Al Mudhnab, Saudi Arabia. AB - In June 1992, 19 cases of typhoid fever were reported from the town of Al-Mudhnab town (population 10,000) in the Qassim region of central Saudi Arabia. Ten of the cases were females (53%), and these were clustered by onset in a 15-day period. Cases in boys followed the girls and were more dispersed over time. Cases included 14 school-age children, four pre-school children, and the wife of a girls' school bus driver. The attack rates (AR) did not differ between boys' and girls' schools and ranged from 5.5 to 15.87 per 1,000 in six different schools. All patients except one lived within two blocks in one quarter of the town. Typhoid fever was associated with attending a school potluck dinner (OR = 5.31; 95% CI 1.02, 28.85) or eating food prepared for that dinner (OR = 6.95; 95%; CI 1.40, 36.61). Cake with cream topping from the dinner was kept overnight at room temperature and served to girls on the bus the next day. The OR of eating leftover cake on the school bus and becoming ill was 11.04 (95%; CI 1.59, 95.27). The localization of this outbreak by time and place and by the age groups affected, supports the conclusion that the outbreak was foodborne. The most likely food was cream topping on the cake. Lessons on proper food handling and storage should be included in home economics classes. PMID- 7498006 TI - Bacterial agents causing chronic suppurative otitis media. AB - Ear swabs from 350 patients with chronic otitis media attending different orthorhinolaryngological clinics at different hospitals and health centres in Benin City and Ekpoma in Edo State were screened for the presence of bacterial agents of chronic otitis media. Results revealed the presence of 19 different species indicating polymicrobial infections. Species isolated comprised Staphylococcus aureus (33.6%), Pseudomonas aeruginosa (19.3%), Proteus mirabilis (17%), Alcaligenes faecalis (6.2%) and Klebsiella aerogenes (4.3%). Others included Escherichia coli (3.3%), Proteus rettgeri (2.8%), and Staphylococcus epidermidis (2.2%), Klebsiella pneumoniae, Proteus vulgaris, Acinetobacter spp, Proteus morgani, Haemophilus influenzae, Providencia spp, Streptococcus pyogenes, Streptococcus pneumoniae, Streptococcus faecalis, non-haemolytic streptococci and Diphotheroids, each accounted for less than 2% of isolates. The study also showed a higher prevalence of chronic otitis media among males (55.7%) than females (44.3%). Cases of chronic otitis media were highest among the age groups (0-5 years) with a prevalence rate of 50% and least among the 6-10 year age group with a prevalence rate of 14.9%. Antibiogram of isolates revealed marked sensitivities (over 90% of the isolates) to ciproxin, tarivid, rocephin and fortum whereas over 70% were resistant to penicillin and ampicillin. Results have indicated that Staphylococcus aureus, Pseudomonas aeruginosa and Proteus mirabilis are leading bacterial agents of otitis media and highlights the high risk involved in the use of penicillin, ampicillin, streptomycin, tetracycline, chloramphenicol, erythromycin, cloxacillin and septrin in the management of chronic otitis media in our locality. PMID- 7498008 TI - Analysis of the results of routine lumbar puncture after a first febrile convulsion in Hofuf, Al-Hassa, Saudi Arabia. AB - Cerebrospinal fluid (CSF) was analysed to determine a lumbar puncture (LP) yield for meningitis in 95 children who presented with their first febrile convulsions between July 1993 and June 1994. There were 52 males and 43 females aged six months to six years with a mean age of 21.9 +/- 13.0 months at presentation. 87(91.6%) had simple febrile convulsions (SFC) while the remaining 8(8.4%) had complex febrile convulsions (CFC). The majority of the subjects presented with a sudden onset of convulsions that were preceded by a day or two history of fever, coryza, cough and respiratory distress while others had their convulsions preceded by fever and passage of bloody stools. The LP yield for meningitis in this series was 6.3%. The CSF analysis revealed six cases of meningitis comprising an eight month old infant with Haemophilus influenzae type B (HIB) meningitis, two partially treated pyogenic meningitis and three aseptic meningitis. All of them had presented with febrile convulsions without signs of meningeal irritation. Excluding aseptic meningitis from this series, a 3.1% LP yield for pyogenic meningitis is significant enough to recommend continued performance of LP in children with first febrile convulsions, especially if under the age of eighteen months. PMID- 7498009 TI - Bedfont Scientific Limited has introduced a revolutionary, purpose-designed inhaled Nitric Oxide therapy monitor called the NOXBOX. PMID- 7498010 TI - Serum calcium, inorganic phosphate and alkaline phosphatase activity in lactating females. AB - Serum calcium, inorganic phosphate and alkaline phosphatase activity were determined in 60 non-lactating and 40 selected lactating females following paturition till the 16th week of lactation at fortnightly intervals. Serum calcium showed a progressive increase from 8.5 +/- 0.1 mg/dl in first week rising to 10.04 +/- 0.4 mg/dl in the 16th week of lactation. Serum albumin and inorganic phosphate were relatively constant up to the sixteenth week postpartum. Serum alkaline phosphatase was highest immediately after paturition, it declines slightly and rises but not above reference range. Serum calcium in the lactating females were usually above hypocalcaemic level and it is thought that dietary calcium was adequate in the lactating females. PMID- 7498011 TI - Clinical evaluation of Norplant in Kumasi, Ghana. AB - A non-comparative study of the Norplant contraceptive subdermal implant system was conducted in Kumasi, Ghana. A total of 100 acceptors were enrolled in the clinical trial, which began in 1987. There were no pregnancies during the first two years of use. Only nine post-insertion medical problems were reported during the first two years of use. One woman in this study requested removal for menstrual pattern changes through two years of use. As assessed through user satisfaction questionnaires administered at approximately six months of use, there has been an overall positive user experience with Norplant in this study. Although the sample size in this study is small, our results seem to be consistent with other studies and it appears that Norplant offers an effective, safe and acceptable method of contraception for Ghanaian women. PMID- 7498013 TI - Percutaneous nephrostomy using hospital modified catheters for cost containment. AB - This paper describes our experience of percutaneous nephrostomy (PCN) using self fabricated catheters. Our effort is to restrain the cost of this common procedure, and at the same time, deliver results comparable to other studies in the literature. In 152 patients, 175 PCN were performed for various indications using simple tools for "pulling catheters". All nephrostomy catheters were fabricated from commercially available 7.2 F polyethylene catheter material. Our success rate of 96%, major and minor complication rates at 2.3% and 6.3% respectively, compare favourably with the reported series. The variable cost of nephrostomy, in our cases, was less than one-third compared with that using commercially available nephrostomy sets. Use of self-fabricated nephrostomy catheters is a cost-effective way of performing nephrostomy especially in environments where health maintenance resources are scarce, health insurance is not freely available, and patients bear the cost of the treatment. This is particularly relevant to the developing countries where PCN is frequently needed but the nephrostomy sets are not easily available. PMID- 7498012 TI - Procured abortion in Ilorin, Nigeria. AB - A prospective study of the maternal mortality and morbidity and other related social problems among 144 cases of procured abortion in Ilorin, Nigeria over a 24 month period is presented. A mortality rate of 90.3 per thousand procured abortions was recorded. Genital sepsis, haemorrhagic anaemia, gut injury, uterine perforation and vesico vaginal fistulae (VVF) were encountered. Poor referral system, late presentation, poor blood transfusion services and inadequate availability of drugs had adverse effects on the patients. The implications (the menace and frequency) of these and possible measures like improving the literacy level, the moral standards, contraceptive practice and family life education (sex education) are discussed. PMID- 7498014 TI - Current drug therapy of systemic mycoses: a review. AB - The frequency of systemic fungal infections have increased significantly in the past decade. Opportunistic systemic fungal infections are increasingly posing difficult management challenges to the clinician. To the aid of the clinician, new systemic antifungal agents have also emerged in the last two decades. To effectively utilise the old and new antifungal agents, a clear understanding of their clinical pharmacology and therapeutic applications is needed. Amphotericin B remains effective therapy in many severe disseminated mycoses including candidaemia and cryptococcosis but problems with toxicity (especially nephrotoxicity), resistances, and non-availability of oral form for long-term maintainance therapy in the immunocompromised patient create important drawbacks. Flucytosine is not effective when used alone and is only used in combination with amphotericin-B in cryptococcal and candidal meningitis. Ketoconazole is available in oral form for systemic antifungal use and is an effective therapy for endemic mycoses, dermatomycoses and oropharyngeal candidiasis. However, it is not satisfactory therapy for deep seated mycoses in the immunocompromised patient. Itraconazole is a new triazole antifungal with better pharmacokinetic profile than Ketoconazole and is currently the drug of choice in most non-life threatening endemic mycosis and is also useful in some deep opportunistic mycoses. Fluconazole is also new triazole antifungal which has the best pharmacokinetic profile and least incidence of adverse effects among all the systemic antifungal available today. It is a first-line therapy for deep candidal and cryptococcal infections in both the immunocompetent and immunocompromised. It is also a first-line maintainance therapy for these infections in the immunocompromised. PMID- 7498015 TI - Priapism: a historical and update review. AB - Early this century, the aetiology of priapism was solely attributed to "systemic disease and local irritation of lower genital tract and neurologic lesion". Corpora cavernosa incision described by Young was the only form of treatment. However, the aetiology, diagnosis and treatment options for the management of priapism and prolonged erections have evolved significantly in the past several years. Before the use of pharmacological agents for the production of erections, idiopathic priapism became the most common aetiology. Causes of priapism from newer psychotropic medications such as trazodone to intra-cavernosal injection therapy with pharmacological agents have increased the number of patients with priapism presenting to the urologist. The management of priapism has remained controversial and has perplexed and continued to frustrate many urologists. A recent and more thorough knowledge of the pathophysiological basis of priapism and the clear differentiation between the low flow veno- occlusive priapism and high flow arterial priapism have significantly improved the diagnostic protocol for patients with priapism. Colour doppler ultrasound evaluation and cavernosal blood gas determinations have become mandatory and greatly improved specific diagnosis. Priapism must be considered a urological emergency and early surgical intervention with corpus cavernosum aspiration and pharmacological lavage with normal saline alpha-adrenegic agonists should be instituted immediately. This will avoid the risk of erectile impotence with considerable medico-legal consequences. Precious time must not be wasted in the older unproven conservative methods including hot and cold water enemas, and vigorous prostatic massage. PMID- 7498016 TI - 'Telephillic syndrome' in pattern and photosensitive epilepsy: report of three cases. AB - The incidence of 'telephillic syndrome' (impulsive attraction to television), in three photosensitive epileptic patients who were referred to our neurophysiology clinic because they were having seizures is reported. Two were females, aged fourteen and eight respectively, one was a male aged twenty-five years. All the patients had a family history of epilepsy. The age range of onset of photosensitivity was 3.5-12 years and the most common seizure inducing factor was flickering light from the television (TV) viewing. They were all 'telephillic'. The predominant seizure type found in these patients was tonic-clonic seizure. Comprehensive electroencephalographic (EEG) investigation was carried out including hyperventilation. The basic EEG showed abnormal spike and wave activity on eye opening and eye closure in all the patients. Intermittent photic stimulation (IPS) evoked occipital spikes and photoparoxysmal response (PPRs) at a photosensitivity range of 758 flashes per second (fps). Pattern sensitivity test using a Gratings pattern revealed a sensitivity range of 1.03-3.5 cycles per second (cpd). And the predominant area of brain activity was the right mid-post temporal regions. Binocular stimulation offered no protection in one patient. PMID- 7498017 TI - Problems in the treatment of kala-azar: case report. AB - Frequent relapses after treatment for visceral leishmaniasis and apparent parasitological cure is not commonly reported. Seven year old boy who relapsed four times with clinical and parasitological evidence of the disease at each two months follow-up period is presented. He had Leishimania donovani Kenya strain. After treatment, review would be after two months, six months and twelve months periods. Splenic aspirates were routinely done weekly and on the last day of each treatment. The drugs administered for varying periods included intravenous sodium stibogluconate 20 mg/kg daily, P20 in combination with allopurinol 21 mg/kg three times daily, and Pentamidine 4 mg/kg three times weekly and antituberculous drugs. The presence of abundant extra cellular leishmania donovani bodies in the bone marrow and possible pulmonary tuberculosis might have precipitated the frequent relapses. It is not clear which of the drugs effected the cure. It was observed that inspite of prolonged antileishmanial drug administration no side effects were noted. PMID- 7498018 TI - Surgical management of infertility: do developing countries need it? PMID- 7498019 TI - Surgical management of female infertility in Ilorin, Nigeria. AB - Three hundred and seventeen female patients with infertility surgically managed and followed up for four years at the University of Ilorin Teaching Hospital (UITH), Ilorin, Nigeria are presented. 45 (14.2%) of these patients were pregnant. A total of 61 pregnancies were achieved in these patients giving an overall pregnancy rate of 19.24% which is comparable to those reported in literature. In the study period, 68.85% full-term gestations, 14.75% pre-term gestations, 11.48% spontaneous abortions and 4.92% ectopic gestations, occurred. As over 50% of female infertility in sub-Saharan Africa is due to tubo peritoneal disorders/pelvic disease and as the desire for children among African women is very strong, the authors plead for the sustenance of surgery, possibly on selected patients, in the management of female infertility even though the success rate appears small. PMID- 7498020 TI - Management of impotence in a developing country. AB - Three hundred and seventy five impotent men attended our Andrology clinic between July 1988 and June 1993. Their ages ranged from 20-89 years. Associated factors were diabetes mellitus, testicular failure, trauma, venous leak and hyperprolactinaemia. We categorised them into subgroups of highly educated men with manual dexterity and financial ability but impairment of manual dexterity, financially able but illiterate men, and illiterate poor with or without manual dexterity. Penile prostheses were implanted in 15; 48 were treated with vasoactive substances; 4 with vacuum devices and two had ligation of venous leaks. However, 297 (79%) men did not get the treatment best suited for their problems and received only supplementary hormones and/or psychological reassurance. In the developing countries, the selection of the appropriate treatment modality for impotence is compounded by considerations of economic, social and educational factors. PMID- 7498022 TI - Hepatitis B viral markers in Bushmen at Schmidtsdrift, South Africa: baseline studies for immunisation. AB - In an effort to control hepatitis B virus (HBV), the prevalence rates of the virus need to be ascertained in different regions or populations. As a baseline for a hepatitis B immunisation programme, the seroprevalence of HBV markers was determined in 400 San Bushmen from a community who were translocated from Northern Namibia to South Africa. HBsAg was detected in 6.8% while 57.0% showed one or more markers of HBV infection. The balance of the population (43.0%) showed no evidence of previous exposure to HBV. The HBV exposure rate of 57.0% is intermediate to high with reference to internationally reported figures and protection of seronegative individuals through immunisation is recommended. Individuals without possible natural immunity against HBV such as those with anti HBc or anti-HBs as a sole marker may provide insight into the circulation of HBV mutants in this community, since these sole markers could be seroconversion in response to a mutant HBV strain. PMID- 7498023 TI - Gastrointestinal tract cancer in association with hepatitis and HIV infection. AB - One hundred and fifty patients with histologically proven gastrointestinal tract (GIT) cancer, 150 patients with a variety of other malignancies and 150 normal subjects were screened for human immunodeficiency virus (HIV) and hepatitis B sero-markers. Only one patient with nasopharyngeal carcinoma proved to be HIV seropositive. Hepatitis B surface antigen (HBsAg) was detected in 18% (n = 26) of the GIT cancer patients, in 16% (n = 24) of the other cancers group and in 12% (n = 20) of the control. There was no significant difference between the three groups (P > 0.1). The HBsAg was detected mainly in patients with primary hepatocellular (25%), gastric (12%), rectal (10%) and colonic carcinoma (8%). Hepatitis B core antibody (HBc AB) was detected in 12% of the GIT cancer patients, in 11% of the other cancers patients and in 13% of the control. In this study, there was no association between HIV, hepatitis B infections and GIT cancer. PMID- 7498021 TI - Factors influencing family planning use in Ilorin, Nigeria. AB - Over a three-year period (1990-1992), the factors influencing the use of family planning among the 7,902 clients attending the Planned Parenthood Federation of Nigeria (PPFN) Clinic in Ilorin were studied. The patients were predominantly females (74.1%). Urban residents (66.1%), educated clients (77.4%), clients aged 25-39 years (73%), and those with three or more children (74.8%) are more likely to use family planning. More Moslems (61%) than Christians (39%) appear to embrace family planning practice in Ilorin. PMID- 7498024 TI - Body temperature and malaria parasitaemia in rural African children. AB - Body temperature was measured and the prevalence of malaria parasitaemia was determined in 198 rural school children aged 6-12 years in a hyperendemic area of southwest Nigeria over a 14 week period spanning part of both wet and dry seasons. Body temperature values in apparently healthy children and in children with malaria parasitaemia were similar with group mean of 37.1 to 37.3 degrees C and with little or no variation in these values with season. The proportion of individual measurements with values > 37.5 degrees C in the two groups were respectively 4.3 and 6%. Despite a seasonal variation in parasite rate, with the highest rates in the wet and the lowest rates in the dry season, there was no significant difference in the proportion of subjects with parasite density > 1000/ul between season. There was also no relationship between parasite density and body temperature. In general, children with parasitaemia < 1000/ul were not pyrexial and less than 2% of all episodes of detectable parasitaemia was accompanied by symptoms of acute malaria. These findings suggest that the presence of malaria parasitaemia has little or no effect on body temperature pattern in a group of rural school children in an endemic area. PMID- 7498025 TI - Perception and utilisation of malaria prophylaxis among pregnant women in Dar es Salaam, Tanzania. AB - Knowledge, perception and utilisation of malaria prophylaxis were assessed among pregnant women attending antenatal care clinic in Dar es Salaam, Tanzania. Of the 301 women interviewed, 71.1% reported having used chloroquine prophylaxis while 28.9% did not. Women with high knowledge of malaria were more likely to use malaria prophylaxis than those with low knowledge. However, there was no significant association between knowledge of malaria and perceived effectiveness of the various methods of malaria control. Chloroquine side effects and perceived lack of protective effect against malaria were mentioned as causes of failure to use chloroquine prophylaxis. Fear of chloroquine-induced pruritus accounted for the largest proportion (49.4%) of women who reported having failed to use chloroquine prophylaxis. Occurrence of malaria episodes was reported to be similar among users and non-users of malaria prophylaxis probably due to inconsistent use of malaria prophylaxis and reduced chloroquine sensitivity of malaria parasites. It is suggested that, in addition to chemoprophylaxis, pregnant women should be encouraged to use bednets in combination with mosquito repellents throughout the course of pregnancy. PMID- 7498026 TI - Incidence of pathogenic microorganisms in clinical specimens from hospitals in south-western Nigeria. AB - The pattern and frequency of bacterial isolates cultured from 687 clinical specimens from four University teaching hospitals in South Western Nigeria were determined. A total of 707 bacterial isolates were cultured from 18 different clinical sources giving an average of 1.03 bacteria per sample identified in this collection. Of these, gram negative rods constituted 78.1% and the rest were staphylococci. The most common organism isolated in urine samples was E. coli (32.1%) whilst S. aureus (28.9%) was the most frequently isolated organism from wounds. An interesting observation was the significant occurrence of Aeromonas hydrophilia and Klebsiella pneumoniae in urine samples. PMID- 7498027 TI - Dietary considerations in designing nutrition intervention programmes in rural areas of Kenya. AB - A comparative study was carried out in three localities in Kenya: Kwale, Mbooni and Lwak in an attempt to identify dietary factors important in designing community based nutrition interventions. The data indicate geographical differences in age at weaning, reasons for termination of breastfeeding, the variety in the diets consumed and the nutrient intake adequacy. However, there was no difference in the duration in breastfeeding and, for children above 6 months (a period when weaning is recommended) rarely was there any food termed as "baby food" but children consumed bits and portions of the family food. In all the localities, there was no statistical difference between households with malnourished children and those without in most of the factors. Households with children admitted in the nutritional rehabilitation centres in the localities were, however, different in most of the characteristics examined. It is concluded that dietary interventions should be targeted to households and not children, mothers' care and nutrition are of paramount importance if the nutrition condition of their children is to be improved and, planning of nutritional interventions should be specific to the districts. PMID- 7498028 TI - Social and psychological aspects of onchocercal skin disease in Nebbi district, Uganda. AB - Onchocerciasis affects 7% of Uganda's population and 1.5 million more people are at risk of infection with Onchocerca volvulus, the nematode that causes the disease. This paper reports the results of part of a multi-centre study whose objective was to determine the prevalence of onchocercal skin disease and its associated psychosocial importance in Uganda. The study employed a standardised clinical dermatological survey method along with the use of structured questionnaires, focus group discussions and key informant interviews. Out of a total of 993 persons examined to determine the prevalence of onchocercal skin lesions 253 persons were interviewed to determine the psychosocial importance of the disease. The results indicate that onchocercal skin disease is associated with a variety of psychosocial, physical and economic effects. The disease also leads to stigmatisation of affected persons and their families. It is suggested that dermatological effects of onchocerciasis should be recognised as an important cause of morbidity in Uganda. PMID- 7498029 TI - Thyroid function in patients with mycetoma. AB - Thyroid function was assessed in seventy two patients with various types of mycetoma. There was no evidence of clinical or biochemical thyroid dysfunction in these patients. The symptoms encountered in some of the mycetoma patients mimic those of hypothyroidism should be attributed to other factors possibly mental depression and apathy. PMID- 7498030 TI - Cancer of the male breast in Zaria, Nigeria. AB - Sixteen Nigerian men presented with advanced breast cancer to Ahmadu Bello University Hospital in Zaria, Nigeria over the 15-year period 1975 to 1989. This comprised 9% of patients with breast cancer seen during this period. They had a median age of 55 years, median symptom duration of 12 months and median tumour size of 8cm; 14 (87.5%) of the patients had Stage IV cancer at presentation. Anaplastic and infiltrating ductal carcinomas accounted for 25% and 56.3% respectively, of the tumour histological type. Mastectomy was the mainstay treatment modality. Only 4 patients came for follow-up, for a median duration of 8.5 months. Overall, this pattern is very similar to the pattern of breast cancer in Nigerian women. PMID- 7498032 TI - Fibrosarcoma of the lung with extrapulmonary manifestations: case report. AB - A 50-year-old female presented with a five months history of recurrent attacks of dizziness, sweatiness, tremors and fainting with loss of consciousness. These were found to be due to hypoglycaemic episodes with blood sugars less than 1 mmol/l and were treated as such. A diagnosis of insulinoma was initially considered, but the patient turned out to have fibrosarcoma of the lung, a rare lung tumour. She also had finger and toe clubbing and features of hypertrophic pulmonary osteoarthropathy. PMID- 7498031 TI - Towards the participation of traditional birth attendants in primary health care in Kenya. AB - This study of 80 traditional birth attendants from Homa Bay District sought their knowledge, functions, and participation in maternal-child health care and family planning. The TBAs were all women with a median age of 50 years. They formed a significant feature of the Luo medical landscape offering a wide range of antenatal, perinatal and postnatal services. The TBAs could become part of the official health care providers offering services, making early diagnoses of difficult pregnancy cases and providing appropriate referrals to health facilities. However, their vocation requires to be enhanced and certain practices such as the use of herbal medicines rationalized through training and research. PMID- 7498033 TI - Cup-and-spill gastric deformity following Hassab's procedure for oesophageal varices: case report. AB - Hassab's operation for selected case of oesophageal varices is believed by many workers to be a very satisfactory procedure. Various complications following this procedure have been reported, but none has reported a case of Cup-and-Spill gastric deformity following this operation. We present such a complication following this procedure on a senior medical colleague with oesophageal varices resulting from a subclinical or previously undiagnosed hepatitis resulting in liver fibrosis. He responded fairly well to conservative treatment and small feed at frequent interval. PMID- 7498035 TI - Cutaneous lesions in hamsters caused by Leishmania donovani promastigotes derived from axenic amastigotes. PMID- 7498036 TI - Cancer of the male genital tract. PMID- 7498037 TI - Management of carcinoma of the penis: a review. AB - Partial penectomy is effective in the treatment of T1 and T2 penile carcinoma with 80% five-year survival rates in the absence of inguinal metastases, and the residual stump is serviceable for upright micturition and sexual function. The use of micrographic surgery first introduced by Mohs in 1941 for small distally located lesions of upto one centimetre diametre achieves results comparable to partial penectomy. Carbon dioxide and Neodymium-Yag lasers have also been used in the treatment of T1 and T2 tumours with 89% five year survival rates. Radiotherapy is also ideal for the treatment of T1 and T2 primary penile carcinoma because failure is corrected with salvage partial penectomy. In T3 disease with extensive local growth total penectomy and removal of scrotal contents followed by perineal reconstruction with scrotal flap is associated with 90% five year survival rates. In advanced T4 tumours with fixed inguinal nodes (N3), hemipelvectomy or hemicorporectomy with combination cytotoxic chemotherapy is considered in selected cases. Various cytotoxic agents like bleomycin, vincristine and methotrexate have been used in adjuvant and neoadjuvant therapy with mixed results. Pre operative radiotherapy is useful in the treatment of patients with metastatic groin lymph nodes of greater than or equal to 4 cm in size. Radiotherapy also provides effective palliation in patients with advanced regional and/or distant metastases. Groin block dissection is commonly performed to treat groin node metastasis and to stage nodal disease in patients with clinically negative groins. The procedure is however associated with significant morbidity resulting in complications such as wound infection and skin necrosis leading to wound breakdown and lymphoedema.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7498034 TI - Management of post-traumatic temporomandibular joint ankylosis in children: case report. AB - Temporomandibular joint (TMJ) ankylosis is a distressing affliction that denies the victim the benefit of normal diet, careers that require normal speech and causes severe facial disfigurement that aggravates psychological stress. Opinions in current literature portray controversy in its management. Hurried clinicians take shortcuts and partially treat symptoms and often make the condition worse. Others ignore shortcomings and erroneously prescribe their preferred techniques as absolute indication to the peril of the affected children. Gap arthroplasty if applied in a growing maxillofacial skeleton as in children, causes iatrogenic arrest of facial growth. The intended correction of facial disfigurement is therefore not achieved. Early detection and immediate psychological support by excision and reconstruction of ankylosed TMJ with a costochondral graft, improves patient comfort and rehabilitation when used as a planned part of a comprehensive therapy. This paper questions prior varied opinions and discusses scientific baseline considerations in management of long standing TMJ ankylosis in children in relatively affordable circumstances. PMID- 7498038 TI - Childhood teratoma in Zaria, Nigeria. AB - Eighty-eight teratomas of diverse sites were seen and managed in our institution between January 1972 and December 1991, in children between the ages of one day and 10 years. Forty seven (53%) tumours were in the sacroccoccygeal region, 17 (19.3%) were of gonadal origin (16 ovarian and one testicular), 13 (15%) were cervical, 4 retroperitoneal. The rest were teratomas of rarer sites. The female/male ratio for various sites were:-sacroccoccygeal teratoma (SCT) 4:1, gonadal 16:1, cervical 1:3, retroperitoneal was seen in 4 patients, all females and for rarer sites 1:1. Overall ratio was 3:1. There were 61 infants and 27 older children. Four SCTs, three of ovarian origin, one cervical and the testicular teratoma were malignant giving an overall malignancy rate of 9%. Serum alpha-foeto protein was not consistently estimated, but was elevated in those patients with malignant teratomas in whom it was sought. PMID- 7498039 TI - Testicular cancer in Nigerians. AB - This is a report of prospective study of eight patients with testicular tumours seen at the Urology Unit of the Lagos University Teaching Hospital over a five year period (1979-1983). The mean age was 32.7 years. Four patients (50%) had germ cell tumours including embryonal carcinoma 25%, seminoma 12.5% and malignant teratoma undifferentiated (MTU) 12.5%. The seminoma in this group originated from a testis which was previously undescended but brought into the scrotum at six years of age. The other four patients (50%), had non germ cell tumours. Two of these patients (25%) had paratesticular tumours including rhabdomyosarcoma of paratesticular adnexae and liposarcoma. One (12.5%) had adenomatoid tumour of the epididymis while the last patient (12.5%) had malignant fibrous mesothelioma of the tunica vaginalis. This study reaffirms the fact that testicular tumours are rare in blacks and that Nigeria has the lowest incidence reported at 0.1 per 100,000 per annum. PMID- 7498040 TI - Childhood leukaemia: experience at King Fahd Hospital, Saudi Arabia. AB - One hundred and twenty eight children suffering from leukaemia were admitted to King Fahd Hospital of the University in the Eastern Province of Saudi Arabia during the period of January 1982-December 1992. Acute lymphoblastic leukaemia was the most common whilst chronic granulocytic leukaemia the least common among the studied group. PMID- 7498041 TI - Oral tumours and tumour-like conditions in Kenya: I. Histological distribution. AB - A histological analysis of 3141 cases of oral tumours and tumour-like lesions diagnosed at Kenyatta National Hospital between 1978 and 1992 has been carried out. Squamous cell carcinoma, salivary gland tumours and lymphoid tumours form the majority of lesions. In general, the histological distribution of the oral tumours do not differ significantly from those reported in non African series save for the Burkitt's lymphoma. PMID- 7498042 TI - Periodontal status following trapezoidal and semilunar flaps in apicectomy. AB - Apicectomy is offered where routine endodontics cannot resolve periapical inflammation. This study compared the influence on periodontal attachment level of two surgical procedures, the trapezoidal (TF) and the semilunar (SF) flaps. Twenty patients aged 16-44 years (mean 23.2 years), were randomly assigned to either flap procedure. Prior to surgery the patients received periodontal prophylaxis and oral hygiene instruction. Records were made of gingival indices, pocket depth and the distance from the cemento-enamel junction to the bottom of the gingival pocket. The measurements involved buccal surfaces of 13 to 23. The TF flap extended from 14 to 24 between the interdental papillae, whereas the SF flap was carried in a semi circle from 14 to 24 about 2mm from the attached gingiva. The surgical wounds were closed with black silk sutures. Antibiotics and analgesics were prescribed. Sutures were removed after one week. On recall after 6, 12 and 24 weeks the same measurements as before surgery were made. No statistically significant change was observed in pocket depth or attachment level between TF and SF flaps (p > 0.05). However, the TF flap produced less noticeable scarification than the SF flap. PMID- 7498043 TI - Oral tumours and tumour-like conditions in Kenya: II. Age, sex and site distribution. AB - This paper presents the age, sex and site distribution of oral tumours and tumour like conditions in Kenya. It confirms some important points. First, there is an overall lower mean age for oral tumours in African series compared to non African series. Secondly, sex bias in the distribution of oral tumours is less marked for most tumours in Africans. Thirdly, there is difference in site distribution and oral tumours between Africans and non-African populations. PMID- 7498044 TI - Histology of gastric antrum in intestinal-type gastric adenocarcinoma from Asir Central Hospital, Saudi Arabia. AB - The objective of this study was to look at non-neoplastic, precancerous histologic changes in the antrum of 49 resected stomachs with intestinal-type adenocarcinoma. Chronic superficial gastritis was seen in all 49 cases. Focal atrophic gastritis was seen in 32/49 cases (65.3%). Helicobacter pylori bacilli were seen in 41/49 cases (84%) and intestinal metaplasia in 39/49 cases (79.6%). 42/49 cases (85.7%) with chronic superficial gastritis showed non-active quiescent chronic inflammation. None of the 49 cases showed early gastric cancer, i.e. adenocarcinoma restricted to the gastric mucosa. The progressive changes of longstanding chronic superficial gastritis, focal atrophic gastritis and intestinal metaplasia were seen in all cases studied (individually or in combination). Knowledge of these precancerous lesions and their prevalence is fundamental to controlling gastric cancer (early detection) and eventual prevention. PMID- 7498046 TI - Acarboxy prothrombin (PIVKA II) as a tumour marker for hepatocellular carcinoma and other liver diseases. AB - The clinical usefulness of plasma abnormal prothrombin, defined as protein induced by vitamin K absence or antagonist II: (PIVKA II) as a tumour marker for hepatocellular carcinoma (HCC) and other liver diseases has been evaluated. PIVKA II concentrations were determined using an enzyme-linked immunosorbent assay (ELISA) with monoclonal antibody that reacts with PIVKA II but does not cross react with normal prothrombin. Seventy four patients (74%) out of 100 with HCC had abnormal PIVKA II levels above 0.5 AU/ml (median = 3.4 AU/ml). The level was above 1.0 AU/ml in 66 (66%) of the patients. In contrast the level of PIVKA II was low in patients with bilharzial periportal fibrosis (median = 0.09 AU/ml), patients with liver cirrhosis (median = 0.13 AU/ml), patients with hepatitis (median = 0.025 AU/ml), and essentially undetectable in all the 34 controls. The diagnostic ability of serum alphafoetoprotein (AFP) was also evaluated in these patients. AFP alone can diagnose 51% of the HCC cases. Of the remaining patients with low or negative AFP levels (65%) can be diagnosed using PIVKA II. Abnormal prothrombin is a potential marker for the laboratory diagnosis of hepatocellular carcinoma. PMID- 7498047 TI - Prevalence of viral infections in Mozambican refugees in Swaziland. AB - The seroprevalence for antibodies to HIV-1, HTLV-1, hepatitis B virus (HBV), hepatitis C virus (HCV) and hepatitis E virus (HEV) were determined in a large group of Mozambican refugees living in Swaziland. Serum samples were collected from a total of 398 refugees located in the two camps (Ndzevane and Malindza). The prevalence for antibodies in the two camps were as follows: Ndzevane: 1.2% (HIV-1); 2.8% (HTLV-1); 0.3% (HCV); 4% (HEV) and 66% for any HBV marker. Malindza: 10.8% (HIV-1); 5.4% (HTLV-1); nil (HCV); 2% (HEV) and 65.7% for any HBV marker. The difference in the HIV-1 seroprevalence between the two camps was statistically highly significant. The phenomenon is possibly related to the location of the Malindza camp in the northern most populous area of Swaziland, resulting in more frequent contact between refugees and the local Swazi population. PMID- 7498048 TI - Costs, resource utilisation and financing of public and private hospitals in Uganda. AB - A descriptive cross-sectional, retrospective cost analysis was conducted in a public and private hospitals in Arua district, Uganda with the aim of assessing the resource use, costs, and financing of those hospitals. It employed the "full costing" approach to hospital costing. The results showed that most of the government recurrent expenditure on health facilities in the district went to the public hospital, while most of the revenues at the private hospital was from donations. The distribution of costs by category of inputs showed that materials and supplies costs were highest in both hospital, followed by employee costs. The administration, pharmacy and store had the highest cost at both hospitals. The cost of the kitchen was very high in the public hospital. The ratio of the unit costs of an out patient visit to an inpatient day and to the unit cost per case treated was 1:1:6 in the private and 1:7:46 in the public. This indicates the relative low cost of public hospital outpatient service and the relative high cost of the outpatient in the private hospital. The employee costs and the drug costs were the most important components of the unit costs in the OPD at both hospitals while in the inpatient wards, besides the above mentioned components, there were also the kitchen, blood bank (public) and theatre in both hospitals. It is concluded that hospitals should look for ways of increasing the level of recurrent funding available to them by increasing the efficiency of their operations and by mobilising additional resources from other sources other than government. PMID- 7498045 TI - Seroprevalence of human immunodeficiency virus infection and its association with syphilis seropositivity among antenatal clinic attenders at Debretabor Rural Hospital, Ethiopia. AB - Sera collected for syphilis screening from pregnant women attending antenatal clinic at Debretabor rural hospital, north west Ethiopia were tested for HIV infection with the objective of assessing HIV prevalence and the relationship between syphilis and human immunodeficiency virus (HIV) seropositivity. A total of 270 pregnant women who visited antenatal clinics from June to September, 1994 were included in the study. Socio-demographic characteristics of the women were retrieved from antenatal care cards. Sera were screened for syphilis and HIV infection by VDRL and ELISA tests respectively. Twenty nine pregnant women (10.7%) were found to be HIV positive. Those living in other villages and rural towns appeared to have a low risk (odds ratio 0.3, 95% confidence interval 0.1 0.8) of acquiring HIV infection when compared to those living in Debretabor town. Women who were seropositive for syphilis were twice as likely to be positive for HIV. Prevention of HIV transmission to women and infants is a priority health and social concern. Prevention and early recognition and treatment of syphilis and other sexually transmitted diseases (STDs) will be important factors in the prevention and control of HIV transmission. PMID- 7498049 TI - Trends in the evaluation of abnormal uterine bleedings with the introduction of manual vacuum aspiration. AB - In a 12-month period, a total of 507 diagnostic curettages were performed with manual vacuum aspiration (MVA) and sharp metallic curettage (SMC), at the Gandhi Memorial Hospital (GMH). Of these, 406(80.1%) and 101(19.9%) were managed with MVA and SMC, respectively. The bulk of the study population were married, of urban residence, paying and treated on ambulatory basis in the minor operation theater without anaesthesia and analgesia. There were significant differences in the gravidity, parity and abortion frequency of the groups (P < 0.05). The mean ages for MVA and SMC were 35.9 years +/- SD 7.6 years and 36.5 years +/- SD 8.5, years respectively (P < 0.05). The majority of the study population that were subjected for histopathological evaluation of the corpus uteri were in the age group of 35-39 years and accounted for 137 (27.0%) with a cumulative frequency of 66.1%. The service providers handled most of the cases with MVA (P > 0.05) and the indications were mainly for abnormal uterine bleeding and endometrial dating. The histological yields were 97.0% for MVA and 80.2% for SMC. The adequacy of the specimen for MVA and SMC were 0.95 and 0.78, respectively. Even if inadequate and non-conclusive are merged together, the adequacy of the specimen would still be 0.91 and 0.74 for MVA and SMC respectively, maintaining the same proportion. Coincidentally, the diagnostic accuracy for endometrial polyp and also for endometrial carcinoma were 0.7 for MVA and 0.3 for SMC. Our results are similar to that of other researchers.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7498050 TI - Haematogram and serum iron status of malnourished Nigerian children. AB - Haematological parameters, total serum iron, and total iron binding capacity (TIBC) concentrations were estimated in twenty protein energy malnourished (PEM) children, five kwashiorkor (K), five marasmus (M), five marasmic-kwashiorkor (M K), and five undernourished (U) aged between one and five years on admission and after 18 days hospitalisation at Obafemi Awolowo University Teaching Hospital Complex. The Hospital diet for K and M-K consisted of 8% protein and 802 calories per litre while that for M and U consisted of 30% protein and 1350 calories per litre. After the period of 18 days of rehabilitation there was no significant (p > 0.05) difference (Student's t-test) in the weight for age. None of the haematological parameters was significantly different after rehabilitation for all the four types of PEM. Of the four types only Kwashiorkor with mean serum iron values of 15.6 +/- 1.51 micrograms/100 ml and 21.17 +/- 0.33 micrograms/100 ml on admission and after rehabilitation respectively was significantly (P < 0.05) different. More attention by the OAUTHC authorities needs to be paid to the management, feeding and diet given to these children so that there will be a greater and more rapid improvement in their rehabilitation. PMID- 7498051 TI - Staphylococcal vertebral osteomyelitis: case report. AB - A 65-year old male Saudi patient presented with rapidly progressive quadriparesis. Lower cervical myelopathy was associated with radiological features of bone destruction, inflammatory disease of the spine and a paravertebral mass. Although tuberculosis and brucellosis are more commonly responsible for this clinical picture in our practice, Staphylococcus aureus was isolated from tissue recovered at surgery. He has been followed up for 12 months and has made an almost complete recovery after surgical decompression of spinal cord and a 2-month course of intravenous flucloxacillin. This case underscores the need for tissue diagnosis in patients presenting with inflammatory paravertebral swellings even in areas endemic for tuberculosis and brucellosis. PMID- 7498052 TI - Update on management of chronic osteomyelitis: report of four cases. AB - Pessimism has existed concerning the cure of patients with chronic osteomyelitis. This has been based on the lack of complete bone and soft tissue debridement in the face of inability to completely cover the resultant wound with adequately perfused tissue. Regional and distant tissue transfers, allow freedom of debridement based on tissue viability rather than on amount of local tissue available to close the wound. Four cases in which regional and distant flaps were used to close defects created after debridement are presented. An overview of reconstruction of such defects is discussed. It is concluded that early management of compound fractures particularly those that require reconstruction will prevent the development of chronic osteomyelitis and thereby undue morbidity. PMID- 7498054 TI - Dietary uptake of superlipophilic compounds by earthworms (Eisenia andrei). AB - The uptake and elimination of three superlipophilic compounds (hexabromobenzene, PCB153, and octachloronaphthalene) after dietary uptake was studied in earthworms (Eisenia andrei). All three compounds were taken up from the food, although they did not significantly accumulate despite their hydrophobicity. Both uptake efficiencies (E) and biomagnification factors (BMF) were low. E varied between 0.70 and 7.5%, while BMF values were all below 0.17. The elimination of the compounds was slow, with elimination rate constants k2 varying between 0.04 and 0.09 day-1. PMID- 7498053 TI - Eccrine acrospiroma in a young African male: case report. AB - This is a report of a rare benign neoplasm of the sweat gland in a 23 year old African male. Clinically, the growth had the appearance of an ulcerated malignant cutaneous neoplasm. It was completely and easily excised at surgery and the patient has remained well without recurrence five years after complete excision. PMID- 7498056 TI - Alterations in protein synthesis in the cyanobacterium Synechococcus sp. strain PCC 6301 in response to Calendula micrantha extract with molluscicidal activity. AB - The response to the extract of the Egyptian wild herb Calendula micrantha, with molluscicidal activity, was examined in the unicellular cyanobacterium Synechococcus sp. strain PCC 6301. Growth and chlorophyll a of the cells were only slightly affected by low concentrations but drastically reduced by high concentrations. The rate of protein synthesis progressively decreased by increasing extract concentration. The cells preferentially induced the synthesis of a limited number of polypeptides in response to the treatment. Among the induced polypeptides were those with apparent molecular weights of 161K (161,000), 96.7K, 93.4K, 85K, 69.9K, 59K, 49K, 45K, 35K, 32.4K, 28K, 24K, 21.7K, 18K, and 16K based on their mobilities in gel electrophoresis. These initial studies suggest that the plant extract exerted certain stress which stimulated alteration in the pattern of protein synthesis in Synechococcus sp. Some of the induced polypeptides are similar to that known to occur in other stresses, especially heat-shock stress. PMID- 7498055 TI - Sublethal sensitivity index as an ecotoxicity parameter measuring energy allocation under toxicant stress: application to cadmium in soil arthropods. AB - Toxic substances may affect the life history of a species in a variety of ways. Different species maintain different priorities in coping with the physiological consequences of toxicant-induced stress. This is expressed by changes in energy allocation to different life-history characteristics which may have great consequences for the response at the population level. In this study the terrestrial invertebrates Platynothrus peltifer (Oribatida), Orchesella cincta, Folsomia candida (Collembola), and Porcellio scaber (Isopoda) were chosen to evaluate species-specific sensitivity of life histories. Effects on reproduction and weight increase under exposure to cadmium in the food were analyzed. The answer to the question of which species is the most sensitive depends on the parameter chosen. A comparison of sensitivity on the basis of sublethal effects showed P. peltifer to be the most sensitive species; on the basis of lethal effects however, the species O. cincta was the most sensitive. This discrepancy between effect parameters resulted in differences between the distance of the concentrations at which lethal and sublethal effects occur for different species. The ratio between the lethal effect concentration and the sublethal effect concentration is called the sublethal sensitivity index (SSI) and is proposed as a parameter expressing maintenance of sublethal functions under toxicant stress. The SSI seems to be a valuable parameter for evaluating the likelihood of population-level effects under toxicant stress. To extrapolate effects found in the laboratory to the field situation, more attention should be paid to the relationships between effects on life-history parameters and effects on population growth. PMID- 7498057 TI - Microalgae and wastewater treatment. AB - The performance of microalgae aquaculture wastewater treatment system predominated mainly by Scenedesmus and Chlorella was assessed. Treatment induced a progressive reduction in both COD and BOD to values below the discharge limits. Different patterns were obtained for removal of phosphorus, nitrogen, and ammonia; however, the algal culture efficiencies reached 100% in their removal by the end of the treatment period. The applied aquatic systems demonstrated percentage reduction of heavy metals in the range between 52.3 and 100% in the batch system and 64.2 and 100% in the continuous system. Wastewater supported algal growth by inducing the incorporation of a significantly higher content of the individual amino acids Asp, Thr, Ser, Glu, Gly, and Tyr, and a markedly higher level of Pro. However, His, Lys, and Arg were markedly reduced compared to their levels in synthetic-medium-grown algae. PMID- 7498058 TI - Xenobiotic biodegradation test using attached bacteria in synthetic seawater. AB - The aerobic biodegradability of aniline, used as reference chemical, has been performed in synthetic seawater with attached biomass in a continuously fed reactor (biofilm chemostat reactor, BCR). Marine bacteria inocula came from local marine fish aquarium filters to limit the geographic and seasonal variations in quality. A pretreatment of these inocula combining 5-microns filtration and centrifugation was used to concentrate bacteria and remove organic carbon contamination of the test. The performances of the BCR were tested in comparison with simple shake flask tests. Among the different variables tested, the ratio S0/X0 (initial concentration of xenobiotic to initial density of the inoculum), the presence of dissolved oxygen, and the hydraulic residence time appear to be the key parameters controlling the length of the biodegradation process. On the other hand, the addition of a cosubstrate (easily biodegradable compound) does not provide advantages. Thus, marine biofilm chemostat reactors with a high density of attached bacteria (around 10(7) cells cm-2) and fed with synthetic seawater plus nitrogen provide good tools for screening biodegradability of chemicals in the marine environment. PMID- 7498059 TI - Influence of the size and source of the inoculum on biodegradation curves in closed-bottle tests. AB - Test variables influencing the results of the closed-bottle test include size and source of the inoculum. The inoculum originates from activated sludge plants maintained at various sludge retention times. The lag period prior to biodegradation, which is influenced primarily by acclimation, decreased with increasing numbers of competent microorganisms. The rate of biodegradation in the closed-bottle test is influenced by the sludge retention time by which the inocula are maintained. Acclimated sludges maintained at low sludge retention times used as inocula gave biodegradation curves with steep slopes, indicating degradation at high rates. Therefore, unacclimated ready biodegradability tests enable only conservative assessments of the biodegradation potential of wastewater purification plants. The results reported here demonstrate that other tests should be used in risk assessment to estimate the full biodegradation potential in wastewater treatment plants. PMID- 7498060 TI - Mn concentration in organs of rats after implantation of battery cathode material. AB - The distribution of Mn, originating from MnO2 in miniaturized batteries, was studied in rats. Forty young male Wistar rats were included in a randomized block experiment with body weight, weight gain, relative organ weight, and the concentration of Mn as dependent variables. Mn containing cathode material was implanted subcutaneously and after 1 month, animals were sacrificed and organs collected. Mn levels were determined by ICPES after destruction of the organic matter using microwave energy. Body weight, weight gain, and relative weight of heart, brain, liver, and kidneys were not significantly altered after implantation of cathode material. However, Mn levels were largely increased in the four organs under study. These high levels did not provoke any of the well known clinical signs associated with Mn intoxication. Effects of Mn in larger farm animals can be considered as insignificant. PMID- 7498063 TI - A comparative study of cadmium, lead, zinc, pH, and bulk density from the Enyigba lead and zinc mine in two different seasons. AB - Soil pH and bulk density, as well as the concentrations of cadmium (Cd), lead (Pb), and zinc (Zn) in soils and these metal levels in the leaves of cassava (Manihot esculenta) and rice (Oryza sativa), both collected, respectively, in the dry season (December) of 1991 and the rainy season (September) of 1992 from the Enyigba lead and zinc mine, Nigeria, were investigated. Interspecific variations in trace element levels in the plant leaves were detected and may reflect the predilection of these plants to particular elements rather than their variability being due to different climatic conditions, such as precipitation, or other environmental factors. PMID- 7498062 TI - Gramine and free amino acids as indicators of fluoride-induced stress in barley and its consequences to insect herbivory. AB - Barley leaves were sprayed with aqueous NaF, which caused accumulation of fluoride in the foliage, but no visible symptoms were detectable. No significant correlation was observed between foliar fluoride concentration and content of the indole alkaloid gramine after exposure to fluoride levels of 20 to 60 mg F liter 1. Fluoride exposure did not explicitly affect the performance of Carausius morosus or Rhopalosiphum paid on barley. After exposure to fluoride levels of 100 and 200 mg F liter-1, as NaF, fluoride treatment had a significant effect on gramine concentration of the first leaf of barley, being highest at a fluoride treatment of 200 mg liter-1, and there was a slight, but significant positive correlation between the log-transformed foliar fluoride concentration and log transformed gramine concentration of the first leaf. Fluoride treatment increased levels of some individual free amino acids in barley foliage. Exposure of young barley to NaF in aqueous form caused accumulation of fluoride in barley foliage and resulted in increased levels of gramine in the first leaf and levels of some free amino acids in foliage. It is possible that the fluoride-induced concurrent increase in gramine concentration in barley leaves could override the eventual increase in nutritive value to herbivorous insects after fluoride exposure. More detailed biochemical studies of the induction of gramine production are needed to understand the fluoride effects in secondary metabolism of barley. PMID- 7498061 TI - Physiological toxicity of low-chlorine bleached pulp and paper mill effluent on whitefish (Coregonus lavaretus L. s.l.): a laboratory exposure simulating lake pollution. AB - In order to validate previous field observations by the authors on whitefish, Coregonus lavaretus L. s.l., a 30-day laboratory experiment with concentrations (0, 1.3, 2.3, 3.5, and 7 vol%) of bleached kraft pulp and paper mill effluent (BKME) simulating those occurring in a polluted lake was conducted. Chlorine dioxide had almost entirely replaced chlorine gas in the bleaching of pulp. As a consequence, the concentrations of adsorbable organic halogens and chlorinated phenolics (CPs) in BKME were significantly lowered compared to earlier studies. This reduction was also seen in the concentrations of CPs in the bile and CPs and extractable organic halogens in the intestinal lipids: the concentrations were low and did not depend on the dilution of BKME. In contrast, the resin acid content of bile decreased with decreasing BKME concentration. The growth of fish was speeded up in all BKME concentrations. However, at the highest BKME concentration (7 vol%) the increase was lowest. The induction of hepatic ethoxyresorufin O-deethylase (EROD) activity revealed strong dose-response relationship with BKME. At 3.5 vol% BKME (corresponding to a distance of 3.3 km from the mill sewer in the field) the EROD activity increased 12-fold. There was a tendency for lower activity of uridinediphosphate glucuronosyltransferase in the liver, but the decrease (34%; P < 0.05) was statistically significant only at 7 vol% BKME. The activity of liver glutathione S-transferase remained unchanged. All dilutions of BKME significantly depressed the concentrations of plasma immunoglobulin M (IgM). Erythrocytic concentrations of nucleotide triphosphates decreased and of sodium increased as the BKME concentration increased. Also some other blood parameters (hematocrit, hemoglobin, plasma glucose, and aspartate aminotransferase) were changed in all BKME exposures, although without obvious dependence on effluent concentration. In conclusion, there was a good agreement between field studies and laboratory experiments using BKME concentrations occurring in the field, confirming close or similar causes for responsive toxicity endpoints. PMID- 7498064 TI - Effects of pentachlorophenol and biotic interactions on soil fauna and decomposition in humus soil. AB - In a laboratory experiment, effects of chemical stress (pentachlorophenol, PCP, at concentrations of 0, 50, and 500 mg/kg) and biotic interactions (nematodes in the presence or absence of collembolas and enchytraeids) on the community structure of soil animals and decomposition processes were studied. PCP was strongly adsorbed to humus that contained 65% organic matter. Numbers of fungal feeding nematodes decreased significantly at the highest PCP concentration, while no effects were found in bacterial feeders. There were differences in the numbers of nematodes between different animal combinations, but at the highest PCP concentration, collembolas and enchytraeids had no effect on them. Numbers of collembola Willemia anophtalma were lowered at the highest PCP concentration, although PCP was not acutely toxic at this concentration. The highest PCP concentration was acutely toxic to enchytraeids, and for an unknown reason all of them died in the main experiment. Both ATP content of the soil and soil respiration were reduced at the highest PCP concentration, while no differences were found between animal treatments. Amounts of NH4-N and PO4-P in the soil increased with increasing PCP concentration. It was concluded that in the presence of simple animal communities, harmful chemicals like PCP regulate the community structure of soil animals as well as decomposition and nutrient mobilization. PMID- 7498065 TI - In vivo effects of deltamethrin exposure on activity and distribution of molecular forms of carp AChE. AB - The in vivo effects of the insecticide deltamethrin (DM) on the activity and molecular forms of acetylcholinesterase (AChE, EC 3.1.1.7) were examined in different organs (brain, blood serum, heart, liver, and skeletal muscle) of carp. The chosen exposure conditions were a DM concentration of 2 micrograms/liter in the water (12 +/- 1 degrees C) for 3 days. All the treated fish survived the experiment, though the effects of the treatment were very visible: the fish always turned on their side, and the skin/scales became infected during the exposure to DM. DM did not result in a significant change in the AChE activity in any of the studied organs except the blood plasma, where the exposure resulted in an AChE activity decrease of as much as 20%. The ratio of membrane-bound to salt soluble AChE forms was determined in the control fish. This ratio increased in the sequence heart, skeletal muscle, liver, and brain. The distribution of the AChE molecular forms was studied in the above tissues. The brain and liver contained forms G1, G4, and A12, the heart and skeletal muscle G4, A4, and A12, and the blood serum G1 and G4. The exposure to 2 micrograms/liter DM for 3 days caused hardly any changes in the pattern of the different AChE molecular forms. A small, but significant (P < 0.05) increase in the proportion of the G4 form was observed in the liver, while G1 and A12 decreased by a few percent (but insignificantly). No other tissues investigated exhibited any changes in the distribution of the AChE molecular forms.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7498066 TI - Further development for testing the effects of pesticides on wolf spiders. AB - Based on preliminary guidelines of the "Biologische Bundesanstalt fur Land- und Forstwirtschaft" (BBA, Germany) for testing the effects of pesticides on wolf spiders (genus Pardosa, Lycosidae, Araneae), two test series were carried out with the pesticides Karate (a.i. lambda-cyhalothrin) and a noname product. At start of the test the pesticides were applied onto spiders and a sand substrate with an application apparatus. Thereafter, the mortality, the behavior, and the feeding rate were observed up to 14 days. Ninety-eight percent of the tested individuals belonged to the species Pardosa amentata. The results of the Karate experiments indicated a considerably higher reaction for males than for females. The aggression of the spiders proved to be a stress factor when the vessel was inhabited by more than one spider and caused higher mortality in individuals treated with pesticides. Spiders raised from cocoons in the laboratory were in general less sensitive than animals collected in the field and accustomed to laboratory conditions. Experiments performed with the noname pesticide in different seasons revealed that the sensitivity of the animals was influenced by their age. The parameters of mortality, behavior, and feeding rate proved to provide data adequate for evaluating the lethal and sublethal effects of the substances examined.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7498067 TI - The polychaete Platynereis dumerilii (Audouin and Milne-Edwards): a new species for assessing the hazardous potential of chemicals in the marine environment. AB - The polychaete Platynereis dumerilii (Polychaeta: Nereidae) has been evaluated as a candidate bioassay species for marine ecotoxicity testing. The species conforms with many of the requirements of an ideal bioassay organism in that (i) it is amenable to laboratory culture, (ii) its relatively small size makes it convenient for handling and laboratory exposure studies, (iii) its diet is defined and can be controlled, (iv) it reproduces throughout the year and, using photoperiod manipulation, can be induced to spawn as required, and (v) it has a short life cycle (approximately 3 months at 20 degrees C) making it feasible to study the effects of xenobiotics on chronic endpoints such as reproduction. The components of the life history which have been examined to date include fertilization rate, embryo-larval development, and larval survival. These life stages were evaluated using the reference materials used in the 1991 International Paris Commission (PARCOM) Ring Test (namely, the biocides, Bioban P 1487 and Vantocil IB, and the widely used reference toxicant, 3,5 dichlorophenol). For fertilization rate, the median effect concentrations (1-h EC50 values) were 0.32 mg.liter-1 for Bioban P-1487, 1.92 [corrected] mg.liter-1 for 3,5-dichlorophenol, and 9.66 mg.liter-1 for Vantocil IB. For embryo-larval development, the median effect concentrations (48-h EC50 values) were 0.29 mg.liter-1 for Bioban P-1487, 2.13 mg.liter-1 for 3,5-dichlorophenol, and 4.81 mg.liter-1 for Vantocil IB. For larval survival, the median lethal concentrations (48-h LC50 values) were 0.32 mg.liter-1 for Bioban P-1487, 3.64 mg.liter-1 for 3,5-dichlorophenol, and 10.9 mg.liter-1 for Vantocil IB. These results (all based on nominal values) suggest that, for these reference materials, the early life stages of P. dumerilii are of similar sensitivity to other marine invertebrate species. Together with the amenability of this species to laboratory culture, these data suggest that P. dumerilii has significant potential for use in marine ecotoxicity testing. PMID- 7498068 TI - Expression of hepatic metallothionein messenger RNA in feral and caged fish species correlates with muscle mercury levels. AB - Metallothioneins (MTs) are low-molecular-weight cytosolic proteins that are induced by cellular stress as well as exposure to various heavy metals including mercury. Excessive residues of mercury have recently been identified in various fish species of the lower Ouachita River system in Arkansas. Fillets of mature largemouth bass (Micropterus salmoides) collected from Woodard Lake, an ox-bow lake of the Ouachita River, possessed muscle residues of mercury ranging from 0.3 to 1.0 ppm (micrograms/g). To assess the usefulness of using MT expression as a biomarker of mercury exposure, livers and fillets were obtained from feral bass of Woodard Lake. Ouachita served as a control site having mercury residues below detection. Analyses using a ribonuclease protection assay with winter flounder MT cDNA revealed that bass had significantly elevated levels of MT mRNA which correlated (r2 = 0.756) with the levels of mercury in muscle fillets. To further explore the water quality of Woodard Lake, 10 juvenile channel catfish were housed in cages and placed where feral collections were made in both sites for 2 weeks. Mercury was not detected in muscle or liver and no significant difference in hepatic MT mRNA was observed. These data demonstrate that MT mRNA expression can be used as a tool to assess exposure to heavy metals and suggest that the elevated levels of mercury in large predatory fish may be due to trophic magnification rather than a single point-source exposure. PMID- 7498069 TI - An evaluation of the aquatic hazard of cumene (isopropyl benzene). AB - Cumene manufacturers were required under a TSCA Section 4(a) test rule to evaluate the aquatic toxicity of cumene to daphnids, rainbow trout, mysid shrimp, and sheepshead minnows. Because of cumene's high volatility (vapor pressure, 3.2 mm Hg at 20 degrees C), all tests were conducted under flowthrough conditions using a proportional diluter system. The 96-hr LC50s for rainbow trout, sheepshead minnow, and mysid shrimp, based on mean measured concentrations, were 4.8, 4.7, and 1.3 mg/liter, respectively. The 48-hr daphnid EC50 was 4.0 mg/liter. Although cumene is considered moderately toxic to aquatic organisms under rigorous laboratory conditions, its volatility and biodegradability greatly reduce its hazard to the aquatic environment. PMID- 7498070 TI - Human root and cord potentials evoked by Achilles tendon tap. AB - Lumbosacral root potentials and sacral spinal cord segmental responses were recorded epidurally in 24 normal adult subjects by tapping the Achilles Tendon with the electronic hammer (mechanical). The averaged responses were also recorded from the same epidural levels after the electrical stimulation of the Posterior Tibial nerve at the popliteal fossa (electrical). While the dorsal and reflexively evoked ventral root responses were easily obtained by the electrical stimulation; by the mechanical stimulation only a late and small ventral root response with two negative deflections could be evoked at the L4-5 or L5-S1 I.V. (intervertebral) epidural levels. The ventral root responses elicited by mechanical stimulation appeared 4.0 msec later than those elicited by the electrical stimulation. This time interval was exactly the same when the latencies of the Soleus reflexes to both stimuli were compared that for the Achilles tendon reflex being 4.0 msec delayed. When the sums of their afferent and the efferent conduction times (to and from Th12-L1 I.V. levels) were extracted from the total conduction times of the Soleus H and Achilles tendon reflex obtained by the EMG recordings, the sacral cord conduction delay was found 1.8 msec for the H-Reflex and 4.1 msec for the Achilles tendon reflex. It was suggested that the twice longer time delay along the sacral cord to Achilles tendon tap might be due to the oligosynaptic linkage of the Achilles tendon reflex or it may result from highly dispersed afferent volley producing a slowly rising wave or excitation that takes longer to reach threshold for the soleus motoneurone discharges. PMID- 7498071 TI - Electrically evoked long-latency responses in human forearm, arm and shoulder muscles. AB - Little is known about electrically evoked long-latency responses (LLR) in muscles other than hand muscles. We obtained LLR in forearm, arm and shoulder muscles by ipsilateral homonymous and heteronymous mixed and pure cutaneous electrical stimulation of the median or ulnar nerve in healthy subjects and in patients with focal unilateral Central Nervous System lesions. Our results point out the important role played by hand afferents in the sensorimotor coordination of multiarticulate upper limb movements. PMID- 7498072 TI - Fast Fourier transform analysis of the masseter muscle EMG during reaction to a warning signal. AB - Frequency changes of the masseter muscle EMG were examined in eight healthy subjects during the reaction time for unilateral jaw biting paradigm. The subjects were instructed to bite on a small piece of wood after the second of two visual signals two seconds apart. In five subjects, the EMG frequencies shifted to lower significantly in both the first (0-1 sec) and the second (1-2 sec) one second periods after the first signal. In two subjects, the EMG frequencies shifted to lower significantly after the first signal in either the earlier or the later period. In one subject, the EMG frequencies did not change. This slowing of the EMG after the first signal might be related to recruitment of larger motor units. This phenomenon must be related to motor preparation for jaw biting. PMID- 7498073 TI - Fibre density in idiopathic ano-rectal incontinence. AB - Fibre density (FD) was measured in the external anal sphincter (EAS) in 54 patients with idiopathic anal incontinence and compared to normative data from a control group of 18 patients. The aim of the study was primarily to establish a diagnostic tool which could give an objective analysis of the anal sphincter muscles. Secondly, to examine whether or not FD was correlated to clinical symptoms of incontinence and manometric measurements from the anal canal. Our data show that the FD was increased in patients with anal incontinence and further that FD was correlated to clinical symptoms of incontinence as well as to manometric parameters of the function of the anal canal. In conclusion, we suggest that FD measurement should be used as a routine in patients with idiopathic anal incontinence. FD values will give an objective estimate of the state of denervation/reinnervation present in the EAS. This is particularly valuable in patients being considered for operative treatment since severe denervation of the EAS generally is correlated to a poor result of surgery. PMID- 7498074 TI - Multimodality evoked potentials in aortoarteritis. AB - Brainstem auditory evoked potentials (BAEPs) have been used to detect early and even asymptomatic brain ischemia. There is a need to detect subclinical brain and/or spinal cord ischemia in patients with aortoarteritis so that ischemic deficits can be prevented by early intervention. We studied multimodality (Visual evoked potentials--VEPs, Somatosensory evoked potentials--SEPs and BAEPs) evoked potentials (EPs) in 10 neurologically asymptomatic subjects with aortoarteritis. EP latencies were normal in all the patients studied. The possible value of EP latencies in detection of early brain and/or spinal cord ischemia in patients with aortoarteritis is discussed. PMID- 7498075 TI - Relation between macro-EMG and muscle Uber conduction velocity. AB - We investigated the relationship between macro-EMG and muscle Uber conduction velocity in the right biceps brachii in 5 normal subjects using a single-Uber macroelectrode and surface array electrodes. The macro-EMG signal and the surface electrode signals were triggered and averaged by the single-Uber action potential. The amplitude and area of the macro-EMG was correlated with the muscle Uber conduction velocity, suggesting that the macro-EMG can serve as a parameter of the size principle in both the clinical and research settings. PMID- 7498076 TI - Functional differentiation within latissimus dorsi. AB - "Functional differentiation" within skeletal muscle refers to the ability of the Central Nervous System (CNS) to control, with a degree of independence, individual subunits of a muscle during a particular muscle contraction. Essentially, the concept of functional differentiation within skeletal muscle suggests an ability of the CNS to selectively activate those segments of a muscle which have the most appropriate line of action for the task as a means of ensuring the muscles efficient utilisation. The present study was undertaken to determine whether functional differentiation was present within the radiate muscle, Latissimus Dorsi, during a series of isometric muscle contractions. Six surface electrode pairs were placed along the origin of the muscle and electromyographic (EMG) potentials were recorded during isometric contractions in various planes and levels of contraction intensity. The EMG waveforms were integrated (IEMG) and then normalised. The normalised EMG waveforms (nIEMG) were then statistically compared to determine if muscle fibres within the detection area of each electrode pair had varied their contribution to the total activity of the muscle, from one isometric contraction to another. Alterations in the contribution of each site were taken to be indicative of functional differentiation. The results of this study indicated that functional differentiation did occur within Latissimus Dorsi. In essence, the most caudal fibres were utilised preferentially when subjects performed adduction from an abducted position of the shoulder joint. Conversely, activity was distributed over the entire muscle when subjects attempted adduction from the anatomical position. A similar situation was observed when subjects performed horizontal extension from either a flexed or abducted position.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7498077 TI - Ataxia-teleangectasia: neurophysiological studies in 8 patients. AB - Eight patients affected by Ataxia-Teleangectasia (AT) were studied with a set of neurophysiological tests. The study was done two times between 2 and 10 years. The most characteristic feature was a severe involvement of the posterior spinal cord: it was associated with a moderate peripheral neuropathy as well as a severe peripheral neuropathy. These data could indicate that the dying back phenomena are not the only pathogenetic basis of the spinal cord degeneration in this disease. PMID- 7498078 TI - Magnetic stimulation of peripheral nerves. Comparison of magnetic stimulation with electrical stimulation. AB - We measured total peripheral motor conduction time by two different methods, magnetic stimulation and electrical stimulation, to reveal which is better in determining actual peripheral motor conduction time. In the upper extremities, the difference between magnetic stimulation and electrical stimulation was within a mean time of 1.39 +/- 0.8 msec, while that in the lower extremities was 2.3 +/- 1.1 msec. In any 4 extremities, total peripheral motor conduction time obtained by magnetic stimulation was significantly shorter than that measured by electrical stimulation employing conventional F-wave response. We conclude that stimulus sites 8.8 +/- 5.1 cm distal to spinal motoneurons in the upper extremities, and 10.5 +/- 5.3 cm distal in the lower extremities were stimulated by magnetic stimulation of the peripheral nerves. Electrical stimulation employing conventional F-wave measurement is superior to magnetic stimulation for actual total peripheral nerve conduction study. PMID- 7498079 TI - The advantages of international cooperation. PMID- 7498081 TI - The Brent Spar saga. PMID- 7498080 TI - Environmental estrogens. PMID- 7498083 TI - The cost of living. PMID- 7498082 TI - EHPnet. PMID- 7498084 TI - New rules for medical waste. PMID- 7498086 TI - Computer recycling takes hold. PMID- 7498085 TI - Selenium secrets. PMID- 7498087 TI - Measuring UV's effects. PMID- 7498088 TI - Menace in the mix. PMID- 7498089 TI - Strategic elements for environmental justice. PMID- 7498090 TI - Assault on the male. PMID- 7498091 TI - Biennial report on carcinogens. A work in progress. PMID- 7498092 TI - An eco-logical way to dispose of waste. PMID- 7498093 TI - Diesel engine exhaust and lung cancer: an unproven association. AB - The risk of lung cancer associated with diesel exhaust has been calculated from 14 case-control or cohort studies. We evaluated the findings from these studies to determine whether there is sufficient evidence to implicate diesel exhaust as a human lung carcinogen. Four studies found increased risks associated with long term exposure, although two of the four studies were based on the same cohort of railroad workers. Six studies were inconclusive due to missing information on smoking habits, internal inconsistencies, or inadequate characterization of diesel exposure. Four studies found no statistically significant associations. It can be concluded that short-term exposure to diesel engine exhaust (< 20 years) does not have a causative role in human lung cancer. There is statistical but not causal evidence that long-term exposure to diesel exhaust (> 20 years) increases the risk of lung cancer for locomotive engineers, brakemen, and diesel engine mechanics. There is inconsistent evidence on the effects of long-term exposure to diesel exhaust in the trucking industry. There is no evidence for a joint effect of diesel exhaust and cigarette smoking on lung cancer risk. Using common criteria for determining causal associations, the epidemiologic evidence is insufficient to establish diesel engine exhaust as a human lung carcinogen. PMID- 7498095 TI - Electric and magnetic field exposures for people living near a 735-kilovolt power line. AB - The purpose of this study was to assess the effect of a 735-kV transmission line on the electric and magnetic field exposures of people living at the edge of the line's right of way. Exposure of 18 adults, mostly white-collar workers, living in different bungalows located 190-240 feet from the line (exposed subjects) was compared to that of 17 adults living in similar residences far away from any transmission line. Each subject carried a Positron meter for 24 hr during 1 workday, which measured 60-Hz electric and magnetic fields every minute. All measurements were carried out in parallel for exposed and unexposed subjects during the same weeks between September and December. During measurements the average loading on the line varied between 600 and 1100 A. The average magnetic field intensity while at home was 4.4 times higher among exposed subjects than unexposed (7.1 versus 1.6 mG, p = 0.0001) and 6.2 times higher when considering only the sleeping period (6.8 versus 1.1 mG, p = 0.0001). Based on the 24-hr measurement, average magnetic field exposure was three times higher among the exposed. Electric field intensity was also higher among the exposed while at home (26.3 versus 14.0 V/m, p = 0.03). Magnetic field intensity among the exposed was positively correlated with the loading on the line (r = 0.8, p = 0.001). Percentage of time above a magnetic field threshold (2 mG or 7.8 mG) was a good indicator to distinguish the two types of exposure.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7498094 TI - Comparisons of estimated human body burdens of dioxinlike chemicals and TCDD body burdens in experimentally exposed animals. AB - Humans are exposed to mixtures of polyhalogenated aromatic hydrocarbons, and the potential health effects of these exposures are uncertain. A subset of this class of compounds produce similar spectra of toxicity in experimental animals as does 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD), and these chemicals have been classified as "dioxins." In this study, we compared the body burdens of dioxins that produce effects in experimental animals to body burdens associated with these effects in humans. Human body burdens were estimated from lipid-adjusted serum concentrations of dioxins, assuming dioxins are equally distributed in body fat and an adult has 22% body fat. The toxic equivalency factor (TEF) method was used to calculate body burdens of dioxins in humans. These calculations included dibenzo-p-dioxins, dibenzofurans, and polychlorinated biphenyls. In the general population, average background concentrations were estimated at 58 ng TCDD equivalents (TEQ)/kg serum lipid, corresponding to a body burden of 13 ng TEQ/kg body weight. Populations with known exposure to dioxins have body burdens of 96 7,000 ng TEQ/kg body weight. For effects that have been clearly associated with dioxins, such as chloracne and induction of CYP1A1, humans and animals respond at similar body burdens. Induction of cancer in animals occurs at body burdens of 944-137,000 ng TCDD/kg body weight, while noncancer effects in animals occur at body burdens of 10-12,500 ng/kg. Available human data suggest that some individuals may respond to dioxin exposures with cancer and noncancer effects at body burdens within one to two orders of magnitude of those in the general population. PMID- 7498096 TI - Biological monitoring of polycyclic aromatic hydrocarbon exposure in a highly polluted area of Poland. AB - Air pollution in Poland and particularly in Silesia is among the worst in Europe. Many coal mines and coke oven plants are located in this area, representing a major source of carcinogenic polycyclic aromatic hydrocarbons (PAHs). We quantitated the PAH exposure level in air samples using personal sampling devices, collected urine samples from the same individuals, and measured 1 hydroxypyrene with high performance liquid chromatography. Samples were collected twice, once in February and once in September. Mean PAH level of samples collected at three different coke oven plants varied from 2.3 micrograms/m3 to 12.3 micrograms/m3; the lowest mean was in September. Mean levels of 0.15 micrograms/m3 (September) and 0.44 micrograms/m3 (February) were noted for the environmentally exposed group. Mean urinary 1-hydroxypyrene varied from 2.45 to 13.48 mumol/mol creatinine at the three coke oven plants. The corresponding variation between the three different environmentally exposed groups in Silesia was 0.41-1.54 mumol/mol creatinine. In the nonindustrialized area, the mean varied from 0.20 to 0.14 mumol/mol creatinine. Seasonal variation was found both at the coke oven plants and in the environmental exposed groups in Silesia. Both PAH levels and 1-hydroxypyrene varied seasonally among coke oven workers and the environmentally exposed group. Our study shows that PAH exposure in the industrialized area of Silesia is high compared to levels in Western Europe. 1 Hydroxypyrene excretion in environmentally exposed individuals in Poland is among the highest in Europe. PMID- 7498098 TI - Risk assessment of environmentally influenced airway diseases based on time series analysis. AB - Threshold values are of prime importance in providing a sound basis for public health decisions. A key issue is determining threshold or maximum exposure values for pollutants and assessing their potential health risks. Environmental epidemiology could be instrumental in assessing these levels, especially since the assessment of ambient exposures involves relatively low concentrations of pollutants. This paper presents a statistical method that allows the determination of threshold values as well as the assessment of the associated risk using a retrospective, longitudinal study design with a prospective follow up. Morbidity data were analyzed using the Fourier method, a time-series analysis that is based on the assumption of a high temporal resolution of the data. This method eliminates time-dependent responses like temporal inhomogeneity and pseudocorrelation. The frequency of calls for respiratory distress conditions to the regional Mobile Medical Emergency Service (MMES) in the city of Leipzig were investigated. The entire population of Leipzig served as a pool for data collection. In addition to the collection of morbidity data, air pollution measurements were taken every 30 min for the entire study period using sulfur dioxide as the regional indicator variable. This approach allowed the calculation of a dose-response curve for respiratory diseases and air pollution indices in children and adults. Significantly higher morbidities were observed above a 24-hr mean value of 0.6 mg SO2/m3 air for children and 0.8 mg SO2/m3 for adults.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7498099 TI - The effect of olestra on breath gas production and faecal microbial counts. AB - OBJECTIVE: To assess the effect of olestra consumption on breath hydrogen (H2) and methane (CH4) production and faecal microbial counts in humans consuming moderate or high fibre diets. A secondary objective was to assess the effect of olestra consumption on health status. DESIGN: Thirty-six-day parallel, placebo controlled, double-blind study with baseline low fibre period (3 g/meal, 8 days) and treatment period (28 days). Four treatment groups: moderate (7 g/meal) or high fibre (24 g/meal), with olestra (24 g) or placebo. SETTING: Queen Margaret College, Edinburgh, Scotland. SUBJECTS: Ninety-seven adult subjects (30 males and 67 females) from the hospital staff and student population participated in the study. Ninety-four subjects successfully completed the study. INTERVENTION: Breath H2 and CH4 production were measured and faecal specimens were obtained for faecal microbial viable counts and direct microscopic cell counts at the end of the baseline period and the end of the treatment period. Standard clinical blood and urine assays were performed. Subjects were questioned regarding adverse events. RESULTS: Olestra demonstrated no significant effect on breath H2 or CH4 production following either moderate or high fibre intake. A trend for lower breath H2 production in the high fibre olestra group was seen. No effect of olestra consumption on faecal microbial counts or health status was observed. CONCLUSION: In normal subjects 24 g/d of olestra for 36 days does not interfere with normal intestinal fermentation of dietary fibre and does not significantly alter gut microflora populations. PMID- 7498100 TI - Effect of winter oral vitamin D3 supplementation on cardiovascular risk factors in elderly adults. AB - OBJECTIVE: A possible role for vitamin D deficiency in contributing to the winter increase in cardiovascular disease mortality was investigated by testing the effect of vitamin D supplementation on blood pressure and other cardiovascular risk factors during winter. DESIGN: Randomised double-blind trial of vitamin D supplementation in winter. SUBJECTS: Men and women, mean age 70 years (range 63 76) recruited from general practitioner age-sex registers in Cambridge (UK). INTERVENTION: 95 people received a single oral dose of 2.5 mg cholecalciferol and 94 received the placebo at baseline interviews during December 1991. Follow-up assessment was 5 weeks later during January 1992. RESULTS: Comparing follow-up with baseline assessment, serum 25-hydroxyvitamin D increased in the treated group and decreased slightly in the placebo group [mean (s.d.) change: 7.2 (+/- 3.8) vs -1.4 (+/- 1.1) ng/ml, P = 0.0001]; while parathyroid hormone decreased in the treated, and increased in the placebo, group [-0.27 (+/- 0.78) vs 0.13 (+/- 0.75) pmol/l, P = 0.0004]. However, the mean change in blood pressure was similar in both groups: systolic -5 (+/- 13) vs -5 (+/- 16) mmHg, P = 0.81; diastolic -1 (+/- 9) vs -1 (+/- 9), P = 0.92; as was the mean change in serum cholesterol [ 0.07 (+/- 0.52) vs -0.05 (+/- 0.60) mmol/l, P = 0.81]. In contrast, the mean change in radial pulse was significantly decreased in the treated group compared with placebo [-2 (+/- 9) vs 1 (+/- 7) beats per min, P = 0.030]. CONCLUSIONS: The failure of vitamin D supplementation to change blood pressure or serum cholesterol suggests that the winter increase in these factors is not caused by decreased vitamin D levels. PMID- 7498097 TI - The E-screen assay: a comparison of different MCF7 cell stocks. AB - MCF7 human breast cancer cells have been studied extensively as a model for hormonal effects on breast cancer cell growth and specific protein synthesis. Because the proliferative effect of natural estrogen is considered the hallmark of estrogen action, it was proposed that this property be used to determine whether a substance is an estrogen. The E-screen assay, developed for this purpose, is based on the ability of MCF7 cells to proliferate in the presence of estrogens. The aim of our study was to characterize the response of four MCF7 cell stocks (BUS, ATCC, BB, and BB104) and determine which of them performed best in the E-screen test. The four stocks assayed were distinguishable by their biological behavior. In the absence of estrogen, MCF7 BUS cells stopped proliferating and accumulated in the G0/G1 phase of the cell cycle; estrogen receptors increased, progesterone receptors decreased, and small amounts of pS2 protein were secreted. Of all the MCF7 stocks tested, MCF7 BUS cells showed the highest proliferative response to estradiol-17 beta: cell yields increased up to sixfold over those of nontreated cells in a 144-hr period. The differences between estrogen-supplemented and nonsupplemented MCF7 BUS cells were due mostly to G0/G1 proliferative arrest mediated by charcoal dextran-stripped serum. MCF7 BUS cell stocks and others showing a similar proliferative pattern should be chosen for use in the E-screen test, or whenever a proliferative effect of estrogen is to be demonstrated. PMID- 7498101 TI - Schoolchildren have similar predicted prevalences of inadequate intakes as toddlers in village populations in Egypt, Kenya, and Mexico. AB - OBJECTIVES: To examine the probability of inadequacy of schoolchild nutrient intakes; to compare the predicted adequacy of schoolchild diets to previously published data for toddler diets and to examine the relative adequacy of the diets of sibling pairs. DESIGN: Food consumption data were measured 2 days per month across approximately one year. Mean nutrient intakes were calculated and compared to international standards using a probability approach. SETTING: Villages in Egypt, Kenya, and Mexico. SUBJECTS: 285 school-age children (7-9 years of age) and 255 toddlers (18-30 months of age) including 94 sibling pairs. RESULTS: Within a country, there was a striking similarity between the nutrients predicted to be inadequate in schoolchild and toddler diets across all nutrients except vitamin B12, which was more likely to be inadequate among schoolchildren. Nutrients of concern for both ages in one or more countries included iron, zinc, calcium, riboflavin, and vitamins B12, D, E, and A. For 94 toddler-schoolchild pairs, siblings at high risk of nutrient inadequacy could be identified from an indicator child with a sensitivity of 43-100% and a specificity of 59-100%. CONCLUSIONS: Nutrient adequacy tends to be a household characteristic for these two ages of children. Nutritional interventions that target the household have a high probability of benefitting multiple age groups of children. PMID- 7498102 TI - Postprandial changes in platelet function and coagulation factors after high-fat meals with different fatty acid compositions. AB - OBJECTIVE: To compare the postprandial effects of three oils differing in their fatty acid composition on platelet aggregation and coagulation. The oils studied were low-erucic acid rapeseed oil (RO, oleic acid 54% of fatty acids), sunflower oil (SO, linoleic acid 64% of fatty acids) and butter oil (BO, saturated fatty acids 62% of fatty acids). DESIGN: The postprandial effects of three fat-loads were followed for 5 h. SETTING: Division of Nutrition, University of Helsinki. SUBJECTS: Twelve healthy female subjects (aged 23-38 years) were recruited among university students and employees. INTERVENTIONS: Postprandial lipaemia was induced by high-fat meals containing fat (RO, SO or BO) 1 g/kg of body weight, skim-milk powder, sugar, strawberries, and water. Each subject ingested each meal in three separate mornings after an overnight fast. The order of the meals was randomised. Blood samples were taken before and 1, 2.5, and 5 h after the test meal. RESULTS: All three test meals similarly affected platelet aggregation in platelet-rich plasma. Aggregation induced by collagen (0.6, 1 and 2.5 micrograms/ml) decreased during the 5-h period after the meals (P = 0.000). ADP induced aggregation did not change during the follow-up period after any meal (P = 0.105-0.483). All fat loads increased factor VII coagulant activity (F VII:C) (P = 0.000), but in plasma fibrinogen concentration (P = 0.155) or antithrombin III activity (P = 0.278) no postprandial changes were found. CONCLUSIONS: These results show that high-fat meals have acute effects on platelet function and F VII:C in healthy women and that these effects are not mediated through the fatty acid composition of the meals. PMID- 7498103 TI - Nutritional status of elderly female nursing home residents; the effect of supplementation with a physiological dose of water-soluble vitamins. AB - OBJECTIVE: To assess the nutritional status of female elderly nursing home residents and to study the effect of dietary supplementation with a physiological dose of water-soluble vitamins. DESIGN: Single-blind, randomised, placebo controlled intervention trial. Daily consumed supplement consisted of fortified fruit juice containing 50 g carbohydrate and 50% of the daily dietary recommendations of water-soluble vitamins. Regular fruit juice was used as placebo. Dietary intake assessment at baseline. Fasting blood sample, anthropometric measurements and medical record at baseline and after 4 and 12 weeks of supplementation. SETTING: Three nursing homes within 40 km of the coordination centre (Department of Human Nutrition, Wageningen Agricultural University, the Netherlands). SUBJECTS: Forty-two female nursing home residents aged 60 years and older, in relatively stable health condition, at least 3 months resident at baseline. In each nursing home, 14 subjects were randomly assigned to the supplement or control group. INTERVENTIONS: Twelve weeks of supplementation. RESULTS: Participants had a poor nutritional status. Dietary supplementation significantly improved the concentrations of thiamin pyrophosphate and pyridoxal 5'-phosphate, increased body weight and decreased serum homocysteine concentration in the supplement group. Plasma vitamin C levels were significantly increased in both supplement and control group. CONCLUSIONS: Even in this heterogeneous population of female nursing home elderly it is possible to improve the nutritional status through dietary intervention with a physiological dose of water-soluble vitamins. PMID- 7498106 TI - Introduction to the special section: the neglected psychological-physical interface. PMID- 7498105 TI - Child malnutrition in poor smallholder households in rural Kenya: an in-depth situation analysis. AB - OBJECTIVE: To investigate the effect of seasonal changes on household food availability on the nutritional status of preschool children from rural households. DESIGN: Households with preschool children were randomly selected from a sampling frame developed from a census list. Data were collected each month or at intervals for 14 months using pre-coded questionnaires. SETTING: The study was carried out in Njoro division, Nakuru district, Kenya. SUBJECTS: Subjects comprised 41 preschool children (22 girls and 19 boys) aged between 18 and 36 months. Households with preschool children were listed from a census conducted earlier. A random sample of 45 subjects was selected, out of which 41 completed the study. INTERVENTION: Food consumption data were collected monthly using the 24-h recall method, and by the 3-day weighed record method during the peak of each season. Subjects were measured once each month, while length was measured at three intervals over the study period. RESULTS: Significant differences in intake of calcium, vitamin A, thiamin and riboflavin were observed between the lean and the post-harvest months, but not with energy, protein, fat, iron, niacin and vitamin C. There were no significant seasonal differences in mean weight changes, which were at the rate of 182 g/month. Mean length increased at a faster rate during the lean season compared to the postharvest months. 51% of the children were stunted during the lean season compared to 28% in the post harvest months. CONCLUSION: The results indicate a seasonal influence on energy and nutrient intake among preschoolers from low income rural households in Kenya. PMID- 7498104 TI - A satiety index of common foods. AB - OBJECTIVE: The aim of this study was to produce a validated satiety index of common foods. DESIGN AND SUBJECTS: Isoenergetic 1000 kJ (240 kcal) servings of 38 foods separated into six food categories (fruits, bakery products, snack foods, carbohydrate-rich foods, protein-rich foods, breakfast cereals) were fed to groups of 11-13 subjects. Satiety ratings were obtained every 15 min over 120 min after which subjects were free to eat ad libitum from a standard range of foods and drinks. A satiety index (SI) score was calculated by dividing the area under the satiety response curve (AUC) for the test food by the group mean satiety AUC for white bread and multiplying by 100. Thus, white bread had an SI score of 100% and the SI scores of the other foods were expressed as a percentage of white bread. RESULTS: There were significant differences in satiety both within and between the six food categories. The highest SI score was produced by boiled potatoes (323 +/- 51%) which was seven-fold higher than the lowest SI score of the croissant (47 +/- 17%). Most foods (76%) had an SI score greater than or equal to white bread. The amount of energy eaten immediately after 120 min correlated negatively with the mean satiety AUC responses (r = -0.37, P < 0.05, n = 43) thereby supporting the subjective satiety ratings. SI scores correlated positively with the serving weight of the foods (r = 0.66, P < 0.001, n = 38) and negatively with palatability ratings (r = -0.64, P < 0.001, n = 38). Protein, fibre, and water contents of the test foods correlated positively with SI scores (r = 0.37, P < 0.05, n = 38; r = 0.46, P < 0.01; and r = 0.64, P < 0.001; respectively) whereas fat content was negatively associated (r = -0.43, P < 0.01). CONCLUSION: The results show that isoenergetic servings of different foods differ greatly in their satiating capacities. This is relevant to the treatment and prevention of overweight and obesity. PMID- 7498107 TI - Pathways linking affective disturbances and physical disorders. AB - Comorbidity of psychological and physical disorders is substantial. This article presents a broad theoretical framework for identifying factors that contribute to and maintain comorbid conditions. The authors propose heuristic models of how co occurrences of psychological and physical disorders are developed and maintained. The models specify biological, behavioral, cognitive, and social pathways that may account for comorbidity. Although the authors' discussion of psychological disorders is limited to the role of affective disturbances (subclinical negative moods as well as mood and affective disorders), the pathways they identify are thought to contribute to co-occurrences of other psychological disorders and physical disease as well. The authors emphasize that pathways linking comorbid states are bi-directional and that operative pathways differ depending on the specific affective response, illness behavior, disease, or disease stage. PMID- 7498108 TI - An archival prospective study of mental health and longevity. AB - The relationship between mental health status and longevity was examined in an archival prospective cohort study (N = 1,103) derived from work begun by Lewis Terman in the 1920s. Degree of psychological maladjustment, cumulatively rated by Terman and his colleagues as of 1950, was found to be related to higher risk of all-cause mortality over a 4-decade follow-up period. The differences among causes of death were nonsignificant, but there was some indication that mental health problems were more strongly related to deaths from injury and cardiovascular disease. The overall relationship was significant for men but weaker for women. The effect was not substantially mediated by alcohol consumption, obesity, or cigarette smoking. PMID- 7498109 TI - The impact of negative emotions on prognosis following myocardial infarction: is it more than depression? AB - This study examine the importance of major depression symptoms, history of major depression, anxiety, anger-in, anger-out, and perceived social support, measured in the hospital after a myocardial infarction (MI), in predicting cardiac events over the subsequent 12 months in a sample of 222 patients. Cardiac events included both recurrences of acute coronary syndromes (unstable angina admissions and survived and nonsurvived MI recurrences) and probable arrhythmic events (survived cardiac arrests and arrhythmic deaths). Major depression, depressive symptoms, anxiety, and history of major depression all significantly predicted cardiac events. Multivariate analyses showed that depressive symptoms, anxiety, and history of major depression each had an impact independent of each other, as well as of measures of cardiac disease severity. PMID- 7498110 TI - Arthritis and perceptions of quality of life: an examination of positive and negative affect in rheumatoid arthritis patients. AB - The utility of measuring both positive and negative affective states for assessing rheumatoid arthritis (RA) patients was examined in 3 independent samples of male and female RA patients (Sample A: 179 women, 48 men; Sample B: 177 women, 24 men; Sample C: 134 women, 38 men). Confirmatory factor analyses of each sample indicated that positive and negative affect constituted separate, negatively correlated factors. The relations among disease variables, coping, and affects were consistent with a model in which coping mediates the relationship between disease variables and positive and negative affect. Patients with higher pain and limitation from RA had higher levels of maladaptive coping, and maladaptive coping was associated with lower positive affect and higher negative affect. Those RAs with higher activity limitation also reported less adaptive coping, which was associated with less positive affect. PMID- 7498112 TI - Predicting outcome of chronic back pain using clinical predictors of psychopathology: a prospective analysis. AB - This study evaluated whether a comprehensive assessment of psychosocial measures is useful in characterizing those acute low back pain patients who subsequently develop chronic pain disability problems. A cohort of 324 patients was evaluated, with all patients being administered a standard battery psychological assessment tests. A structured telephone interview was conducted 6 months after the psychological assessment to evaluate return-to-work status. Analyses, conducted to differentiate between those patients who were back at work at 6 months versus those who were not because of the original back injury, revealed the importance of 3 measures: self-reported pain and disability, the presence of a personality disorder, and scores on Scale 3 of the Minnesota Multiphasic Personality Inventory. These results demonstrate the presence of a psychosocial disability variable that is associated with those injured workers who are likely to develop chronic disability problems. PMID- 7498111 TI - A longitudinal study of life events and metabolic control among youths with insulin-dependent diabetes mellitus. AB - In a study of school-age children with new-onset insulin-dependent diabetes mellitus (IDDM), life stress, metabolic control (glycosylated hemoglobin), and psychiatric and psychosocial variables were assessed repeatedly for up to 6 years. Life stress was defined as the number of undesirable life events and extent of life change necessitated by all life events. In univariate longitudinal analyses, total life change units were related to metabolic control, with a trend for number of undesirable events to be associated with metabolic control. In multivariate analyses, metabolic control was related to life change units, whether the glycosylated hemoglobin was in the 1st year of IDDM, IDDM duration, and the diagnosis of pervasive noncompliance with medical regimen. The relationship of life stress to metabolic control among youths with diabetes in significant but clinically modest and may be partially mediated by serious noncompliance with the medical regimen. PMID- 7498113 TI - Predictors of panic-fear in asthma. AB - Eighty-six asthmatics completed measures of illness-specific panic-fear (i.e., panic-fear in response to symptoms of asthma) and of generalized panic-fear, dyspnea frequency, and catastrophic cognitions about bodily symptoms (the Anxiety Sensitivity Index [ASI] and Agoraphobic Cognitions Questionnaire [ACQ]). Asthma variables (self-report and pulmonary function tests) and cognitive variables (ASI and ACQ) were independently related to illness-specific panic-fear. Regression analyses showed that the cognitive variables predicted significant variance in both panic-fear scales after controlling for the effects of demographic and asthma variables. By contrast, the asthma variables were not associated with generalized panic-fear when the cognitive measures were controlled. In light of the adverse effects of panic-fear on asthma, the authors' results suggest that researchers may fruitfully explore the use of cognitive techniques as an adjunctive treatment for improving asthma outcome. PMID- 7498114 TI - Responses to hypothetical high risk situations: do they predict weight loss in a behavioral treatment program or the context of dietary lapses? AB - Before starting a behavioral weight control program and after 6 months of treatment, 93 participants were asked to listen to 4 scenarios describing typical high risk situations for dieters (family celebration, watching TV, tension at work, argument). The number of coping responses generated, latency of responses, and perceived risk of lapsing were assessed; participants also identified which situation would be most difficult for them. Telephone interviews were conducted during the weight loss program to assess situations surrounding actual dietary lapses. Participants who rated the negative affect situations as most difficult were more likely to lapse in situations involving negative affect; 87% of these participants lapsed in association with negative affect. Moreover, participants who generated coping responses to more of the situations subsequently lost more weight. Thus, baseline performance on a hypothetical high risk task predicts subsequent performances in a behavioral weight loss program. PMID- 7498115 TI - Effects of surgical menopause on psychological characteristics and lipid levels: the Healthy Women Study. AB - The authors examined the effects of surgical menopause, with or without hormone replacement therapy (HRT), on psychosocial functioning and lipid levels in a population-based study of initially premenopausal women. Within 5 years of study entry, 28 women underwent hysterectomy, with 9 retaining their ovaries and 19 having bilateral salpingo oophorectomy (BSO), and with 173 premenopausal women as a comparison group. Women undergoing hysterectomy reported fewer stress symptoms and a more optimistic attitude at follow-up, whereas BSO cases without HRT had lower mean total high-density lipoprotein (HDL) and HDL2, relative to premenopausal women. Subject to limited power, the authors' findings suggest that surgical menopause among middle-aged women does not lead to negative psychological outcomes but that BSO in the absence of hormone use has significant adverse effects on the HDL and HDL2. PMID- 7498116 TI - Relationship of trait anger to resting blood pressure: a meta-analysis. AB - A series of meta-analyses were conducted to assess whether anger is related to essential hypertension. The present review also considered the relevance of the distinction between anger experience and anger expression, the effect of participant selection bias, and the white-coat hypertension effect for the anger blood pressure (BP) association. Anger experience was correlated with elevated BP, but the relationship was small and highly variable. When positive effects emerged, both participant selection and the reliability of BP measurement posed interpretational problems. Persons high in anger are not merely exhibiting elevated BP in response to testing, so a white-coat effect is not evident. Being labeled as hypertensive may contribute to higher anger scores, however. The review suggests lines of future research concerning associations between trait anger and blood pressure. PMID- 7498117 TI - Burden of self-care in seriously ill patients: impact on adjustment. AB - Perceived, but not actual, control over the treatment has been consistently related to better adjustment in chronic illness. This study examined the relationship between actual control over treatment and severity of illness and their influence on depression in a chronically ill population of end-stage renal disease (ESRD) patients. The authors hypothesized that as severity of illness increases, the burden of control over treatment would increase depression. Severity of illness and depression were assessed for 98 ESRD patients. Control over treatment was represented by whether dialysis patients were self administering treatment (high control) or were receiving treatment from the medical staff (low control). Results indicated that for the most severely ill patients, high control over treatment resulted in poorer adjustment. Furthermore, this effect was due in part to how illness interferes with social relationships in seriously ill, self-care patients. PMID- 7498118 TI - Parental education related to adolescent stress-coping and substance use: development of a mediational model. AB - This research tested the relation of socioeconomic status (SES), indexed by parental education, to stress-coping variables and substance use in a sample of 1,391 adolescents. Lower education was related to higher level of adolescent substance use, lower levels of protective factors, and higher levels of risk factors. Mediation analyses indicated that the effect of education on adolescent substance use was mediated through relationships to parental support, academic competence, behavioral competence, negative life events, and friends' substance use. Moderation analyses indicated that adolescents from lower education families were more vulnerable to risk factors but also derived more benefit from protective factors. Multiple-group analysis indicated that moderation was attributable to larger paths in the lower education group for negative events and friends' use. Implications for the relation between SES and health risk are discussed. PMID- 7498119 TI - The determination of nitrite and nitrate in human blood plasma by capillary zone electrophoresis. AB - The determination of nitrite and nitrate in human blood plasma by capillary zone electrophoresis (CZE) is described. Electrophoresis was carried out in a 65 cm long, 75 microns wide silica capillary, using 750 mM sodium chloride containing 5% NICE-Pak OFM Anion-BT (an additive reversing the electroosmotic flow) as running buffer, at a potential of 20 kV, with on-column UV detection at 214 nm. Under these conditions, the two anions migrated in the order of nitrite and nitrate, completely resolved from each other. Forty-one samples of ultrafiltered blood plasma of healthy volunteers were analyzed. The concentrations of nitrite and nitrate were 0.15 +/- 0.07 mg/L and 3.2 +/- 1.6 mg/L (mean +/- SD), respectively, with a linear correlation in the range of 0.1-50 mg/L. The within day and between-day coefficients of variation (CVs) for nitrite and nitrate were lower than 5% and 8%, respectively. The recovery was 97-114%. The estimated plasma volume needed for each determination was 108 nL. PMID- 7498120 TI - Purification of wheat glutenin subunits by preparative acid and two-dimensional electrophoresis. AB - Reduced alkylated glutenin subunits from wheat flour were fractionated by preparative electrophoresis at acid pH. The high molecular weight glutenin subunits (HMW-GS) and some of the low molecular weight glutenin subunits (LMW-GS) were purified by this one-step procedure, whereas the remainder of the LMW-GS, comigrating in the acid system, were purified in a second step by electroendosmotic preparative electrophoresis in the presence of sodium dodecyl sulfate. The quantities of recovered protein were sufficient for biochemical characterization and/or antibody production. PMID- 7498121 TI - Free flow-isoelectric focusing of human cellular lysates as sample preparation for protein analysis. AB - A whole cell lysate of human cells was separated into 80 fractions according to the pI of proteins using free flow isoelectric focusing with carrier ampholytes. The resolution of the process was highly reproducible, with an overlap of fractions of less than 30%. A protein of a faint silver stained spot in two dimensional gel electrophoresis (2-DE) could be enriched, yielding a Coomassie blue stained spot which could be further characterized by protein chemical methods. The enrichment of less abundant proteins from a complex crude cell extract was found to be a suitable tool for sample preparation and enrichment before applying proteins to 2-DE and reversed-phase high performance liquid chromatography. PMID- 7498122 TI - Direct detection of beta-1,3-glucanase in plant extracts by polyacrylamide gel electrophoresis. AB - By using carboxymethyl (CM)-curdlan, a polysaccharide linked with the dye Remazol Brilliant Blue (RBB) as a substrate in polyacrylamide gels, the beta-1,3 glucanase in plant extracts can be detected directly by native polyacrylamide gel electrophoresis. In contrast to the usually used procedures for the detection of glucanases, e.g., colorimetric assay, overlay technique, enzyme activity staining using laminarin as a substrate, this method is rapid and allows both the determination of the activity and the location of the relative position of the multiple forms of beta-1,3-glucanases. PMID- 7498123 TI - Visualization of barley beta-glucan degrading isozymes after gel isoelectric focusing. AB - A simple method to study the polymorphism of barley (Hordeum vulgare L.) beta-(1 3,1-4)-glucanases (specific enzymes of barley beta-glucan hydrolysis) is described. Proteins of a crude extract of germinated barley kernels were separated in an immobilized pH gradient in two pH ranges (pH 3-10.5 and pH 4-7). beta-glucanases were visualized by contact printing with a polyacrylamide gel containing beta-glucan or lichenan. Patterns of beta-glucanases were revealed by staining with Congo Red with resultant clear zones on a stained background. Various conditions of germination, extraction and visualization were investigated. New isozyme bands could be detected and their nature and origin discussed. PMID- 7498124 TI - Group-specific component subtyping in semen and seminal stains by conventional isoelectric focusing. AB - Group-specific component (GC) subtyping in semen and seminal stains was carried out using isoelectric focusing in carrier ampholyte-generated pH gradients and immunoblotting. In serum samples the anodal bands of GC 1F and of GC 1S disappeared by neuraminidase treatment, but in semen samples these bands remained unchanged after such treatment. The GC 2 type in semen exhibited two bands: the main GC 2 band and another fast band which focused at the position of the cathodic band of GC 1F. These seminal GC bands were unaffected by enzyme digestion. Reliable subtyping was possible in seminal stains stored at 4 degrees C for up to 10 weeks, at room temperature for up to 8 weeks, and at 37 degrees C for up to 5 weeks. The GC subtyping by conventional isoelectric focusing after neuraminidase treatment is simple, economical and useful in medicolegal examination of seminal stains. PMID- 7498125 TI - Serine protease inhibitor activity of recombinant squamous cell carcinoma antigen towards chymotrypsin, as demonstrated by sodium dodecyl sulfate-polyacrylamide gel electrophoresis. AB - Squamous cell carcinoma (SCC) antigen was tested, by sodium dodecyl sulfate polyacrylamide gel electrophoresis, for its ability to inhibit the activity of serine proteases, i.e., trypsin, chymotrypsin and elastase. We demonstrated that the serine protease inhibitor (serpin) of SCC antigen is specific for chymotrypsin. Preincubation of chymotrypsin with recombinant SCC antigen inhibited chymotryptic digestion of gelatin and ovalbumin through the formation of sodium dodecyl sulfate-stable complexes. These findings promote understanding of the biological functions of SCC antigen as serpin in the stratification of the normal squamous cells and in the malignant behavior of the tumor cells. PMID- 7498126 TI - Transient bisalbuminemia: separation by isoelectric focusing of human albumin fractions linked to different numbers of benzylpenicilloyl groups. AB - The behavior of modified human serum albumin (HSA) from penicillin-treated patients and in vitro conjugated samples was studied by isoelectric focusing in the absence and presence of 8 M urea. It is shown that samples classified as bisalbuminemic after separation under native conditions display a spectrum of several major bands with decreasing pIs when focusing is done in the presence of 8 M urea. In this spectrum, the distances between two consecutive major bands are as expected from a 2-unit charge difference. Since this result is compatible with the blockage of a lysine residue with a benzylpenicilloyl (BPO) group, each major band can be interpreted as representing an HSA fraction covalently linked to different numbers of benzylpenicilloyl groups. The separation methods presented therefore seem to constitute simple and reliable tools for a detailed assessment of the extent of BPO covalent binding to HSA both in vivo and in vitro. PMID- 7498127 TI - Two-dimensional electrophoresis of proteins: an updated protocol and implications for a functional analysis of the genome. AB - The two-dimensional electrophoresis (2-DE) technique developed by Klose in 1975 (Humangenetik 1975, 26, 211-234), independently of the technique developed by O'Farrell (J. Biol. Chem. 1975, 250, 4007-4021), has been revised in our laboratory and an updated protocol is presented. This protocol is the result of our experience in using this method since its introduction. Many modifications and suggestions found in the literature were also tested and then integrated into our original method if advantageous. Gel and buffer composition, size of gels, use of stacking gels or not, necessity of isoelectric focusing (IEF) gel incubation, freezing of IEF gels or immediate use, carrier ampholytes versus Immobilines, regulation of electric current, conditions for staining and drying the gels - these and other problems were the subject of our concern. Among the technical details and special equipment which constitute our 2-DE method presented here, a few features are of particular significance: (i) sample loading onto the acid side of the IEF gel with the result that both acidic and basic proteins are well resolved in the same gel; (ii) use of large (46 x 30 cm) gels to achieve high resolution, but without the need of unusually large, flat gel equipment; (iii) preparation of ready-made gel solutions which can be stored frozen, a prerequisite, among others, for high reproducibility. Using the 2-DE method described we demonstrate that protein patterns revealing more than 10 000 polypeptide spots can be obtained from mouse tissues. This is by far the highest resolution so far reported in the literature for 2-DE of complex protein mixtures. The 2-DE patterns were of high quality with regard to spot shape and background. The reproducibility of the protein patterns is demonstrated and shown to be thoroughly satisfactory. An example is given to show how effectively 2-DE of high resolution and reproducibility can be used to study the genetic variability of proteins in an interspecific mouse backcross (Mus musculus x Mus spretus) established by the European Backcross Collaborative Group for mapping the mouse genome. We outline our opinion that the structural analysis of the human genome, currently pursued most intensively on a worldwide scale, should be accompanied by a functional analysis of the genome that starts from the proteins of the organism. PMID- 7498128 TI - Separation of heat-stable proteins from Thermus thermophilus HB8 by two dimensional electrophoresis. AB - Thermostable proteins from Thermus thermophilus HB8, an extremely thermophilic bacterium, were separated by two-dimensional gel electrophoresis. About 1200 spots were detected with silver staining on the gel between pH 3 and 10. According to the genome size of T. thermophilus, we consider that more than half of the proteins in the cell are visualized on a two-dimensional gel. Using comigrated standard marker proteins, the molecular weight and isoelectric point of each protein spot were calculated. The average molecular weight and isoelectric point values were estimated to be 30 000 and 5.2, respectively. The average size and isoelectric point of detected protein from T. thermophilus were smaller and more acidic than those from Escherichia coli. After the protein spots had been electroblotted onto a polyvinylidene difluoride membrane and stained with Coomassie Brilliant Blue, the N-terminal amino acid sequences were determined for about twenty protein spots. Few proteins had blocked N-termini. Some spots were identified as proteins whose sequences had been reported previously from T. thermophilus. Others had amino acid sequences homologous with those of various proteins from other organisms. The amino acid sequence information of this report will be useful for obtaining stable proteins and for identifying open reading frames determined from the genome DNA sequence. Considering its small genome size and protein stability, T. thermophilus will be an excellent candidate for studying the molecular biology of an autotrophic living cell at the atomic level. PMID- 7498129 TI - Two-dimensional electrophoretic analysis of down-regulated proteins in human fibroblasts immortalized by treatment with either 4-nitroquinoline 1-oxide or 60Co gamma rays. AB - Cellular proteins were examined by two-dimensional polyacrylamide gel electrophoresis (2-D PAGE) in order to determine the mechanisms of cellular aging and immortalization of human fibroblasts. A total of 10 cell lines were studied, four of which were immortalized. OUMS-24F and SUSM-1 cells were immortalized by repeated treatment with 4-nitroquinoline 1-oxide (4NQO), KMST-6 cells were immortalized by intermittent exposure to 60Co-gamma rays, and IMR-90/SV40 cells were immortalized with simian virus 40 (SV40). None of these immortalized cells were tumorigenic. Four normal human fibroblast cell lines, OUMS-24, AD 387, KMS-6 and IMR-90, and two human cervical cancer cell lines, HeLa and A-431, were also studied. Applying 2-D PAGE, the down-regulation of the identical proteins was observed in the four immortalized cell lines and in the two human cancer cell lines. Their molecular masses were about 80 kDa and the isoelectric points ranged between 5.5 and 6.3. We could not find any information on our proteins from the recent protein data bases. Further characterization of the proteins indicated that they might be phosphorylated cytoskeletal proteins. PMID- 7498130 TI - Theory of the mobility-shift assay of nonspecific protein-DNA complexes governed by conditional probabilities: the HU:DNA complex. AB - Complexes of an 88 bp DNA and the HU protein were studied by both experimental and theoretical electrophoretic mobility-shift analyses. Experimental analysis defined the stoichiometry of binding and estimated an apparent intrinsic dissociation constant (Kd = 1 to 3 x 10(-7) M) for the HU:DNA complexes. The theory of conditional probabilities was applied to the binding of HU to DNA in order to fix the initial equilibrium composition of mixtures to be assayed theoretically by the mobility-shift procedure. Electrophoretic mobility-shift patterns were obtained by numerical solution of a set of simultaneous transport reaction equations, in which the chemical kinetic term is formulated in terms of dissociation of the different DNA:HU complexes in gel cages. The computed patterns simulated the experimental patterns describing the titration of a fixed concentration of an 88 bp DNA fragment with dimeric HU. These insightful results provide guidelines for interpretation of the electrophoretic behavior of systems in which a ligand binds nonspecifically to DNA. In particular, the narrow unresolved zone observed both experimentally and theoretically beyond 50-60% saturation is a reaction zone characteristic of noncooperative ligand-binding governed by conditional probabilities. The discrepancy between the theoretically assigned and experimental values of the intrinsic binding constant is attributed to an HU-induced change in the conformation of DNA. PMID- 7498131 TI - Tris-dependent oxidative DNA strand scission during electrophoresis. AB - The DNA of two Streptomyces species contains site-specific labile modifications. During gel electrophoresis the DNA can undergo Tris-dependent strand scission at the positions of these modifications. Our investigations into the nucleolytic activity which reacts with the modifications implicate a peracid derivative of Tris formed at the anode; the kinetics of production and decay of this activity were followed using both a DNA cleavage assay and a reduced methyl viologen assay to measure oxidant. Anode activation could be chemically mimicked by addition of peracetic acid to Tris buffers. We tested the DNA cleavage activity of several other compounds after anode or chemical activation; we used an analogue of Tris lacking a primary amine group and also several reagents known to promote DNA strand cleavage by amine-catalysis at abasic sites. Anode generation of oxidant could be detected for compounds containing either hydroxyl or carboxyl groups. However, DNA cleavage activity correlated with oxidant formation only for those compounds also containing primary amine groups. These results support a mechanism of DNA strand scission at modification sites via concerted peracid-mediated oxidative and amine-catalysed reactions. The novel finding of Tris-dependent formation of a long-lived reactive oxidant at the anode suggests that this compound is unsuited as an electrophoresis buffer for certain biological macromolecules. PMID- 7498132 TI - Dispersion coefficients of a protein and DNA fragment in polyacrylamide gel electrophoresis as a function of parameters defining the effective gel pore size and particle size. AB - The dispersion coefficient, D', of the representative homogeneous protein, conalbumin, decreases linearly as the polyacrylamide concentration increases from 4 to 14%T (2%C), and varies in a biphasic fashion as %C (Bis) is increased from 2 to 20%, with a broad peak between 5 and 15%C. D' increases linearly with the concentration of the initiator, potassium persulfate, in the range of 0.01-0.15%. D' remains constant when the field strength is varied from 5 to 15 V/cm. A DNA fragment (1857 bp) exhibits a constant D' in 4-6% polyacrylamide (2%C) at a field strength of 1 V/cm, and a linearly increasing D' at 5 V/cm, in analogy to its previously observed behavior in agarose gels. In solutions of uncrosslinked polyacrylamide, the decrease of the D' of conalbumin with polymer concentration is not significantly different from that in 2% N, N'-methylenebisacrylamide crosslinked gels in the range of 4-14%T, while the decrease of mobility with polyacrylamide concentration is much steeper in 2% crosslinked compared to uncrosslinked polymer. Finally, delta (log D')/ delta T was found to be proportional to the retardation coefficient, KR (= -delta (log mu)/ delta T), in polyacrylamide gels. The ratio of -delta (log D')/ delta T over KR increases with field strength in the range of 5-15 V/cm. PMID- 7498134 TI - The influence of proteins on silver staining of nucleic acids following polyacrylamide gel electrophoresis. AB - The influence, on silver staining, of proteins and restriction enzymes in polymerase chain reaction (PCR) products was studied in 12-20% polyacrylamide gels. For small DNA fragments (74, 41 and 33 bp) best results were achieved using 20% polyacrylamide gels. In 12% gels, restriction enzymes and also proteins used for PCR (bovine serum albumin, Taq-DNA polymerase) interfere with silver staining of nucleic acids. PMID- 7498133 TI - Genotyping apolipoprotein B signal peptide insertion/deletion: a comparison of three methods. AB - Three different ways of detecting insertion/deletion (Ins/Del) genotypes of the apolipoprotein B gene encoding signal peptide were compared. Unequal quantities of the two alleles were observed in 50 Serbian subjects. The low abundance allele was clearly visible only when a modified method of silver staining for polyacrylamide gels was used. This method is more reliable for Ins/Del polymorphism genotyping, avoiding misclassifications by conventional ethidium bromide staining of both agarose and polyacrylamide gels. PMID- 7498135 TI - The Dukhin-Deryaguin equation for the electrophoretic mobility in monovalent electrolytes with arbitrary ion mobilities. AB - The Dukhin-Deryaguin equation, used in the modern theory of electrophoresis for the calculation of electrophoretic mobility (EPM) in the region of double layer polarization, is known from literature [4-6] in its form (DD1) valid for the equal mobilities of the cation and anion in solutions of symmetrical electrolytes. Here we describe the other version of this equation (DD2), for arbitrary ion mobilities in 1:1 electrolytes. The EPMs calculated from this latter version are in good agreement with an exact computer solution [12]. The use of DD2 is illustrated in a series of EMP vs. IgC curves, calculated for selected examples of negatively charged lipid membranes. In addition, we describe two simplified versions of DD2, which are valid, respectively, for the high zeta potentials and when the electroosmotic component of ions' fluxes at the charged surface is neglected. Comparing DD2 with DD1 shows that the latter equation results in an error which may exceed the experimental dispersion of EPM values in the absence of specific ion binding. This error is reduced if the counter-ion binding is not small; hence, DD1 may also be used in some cases for solutions with arbitrary ion mobilities. PMID- 7498136 TI - Single-step electrotransfer of reverse-stained proteins from sodium dodecyl sulfate-polyacrylamide gel onto reversed-phase minicartridge and subsequent desalting and elution with a conventional high-performance liquid chromatography gradient system for analysis. AB - Isolation of proteins from polyacrylamide electrophoresis gels by a novel combination of techniques is described. A given protein band from a reverse stained (imidazol-sodium dodecyl sulfate--zinc salts) gel can be directly electrotransferred onto a reversed-phase chromatographic support, packed in a self-made minicartridge (2 mm in thickness, 8 mm in internal diameter, made of inert polymeric materials). The minicartridge is then connected to a high performance liquid chromatography system and the electrotransferred protein eluted by applying an acetonitrile gradient. Proteins elute in a small volume ( < 700 microL) of high-purity volatile solvents (water, trifluoroacetic acid, acetonitrile) and are free of contaminants (gel contaminants, salts, etc). Electrotransferred proteins were efficiently retained, e.g., up to 90% for radioiodinated alpha-lactalbumin, by the octadecyl matrix, and their recovery on elution from the minicartridge was in the range typical for this type of chromatographic support, e.g., 73% for alpha-lactalbumin. The technique was successfully applied to a variety of proteins in the molecular mass range 6-68 kDa, and with amounts between 50 and 2000 pmol. The good mechanical and chemical stability of the developed minicartridges, during electrotransfer and chromatography, allowed their repeated use. This new technique permitted a single step separation of two proteins unresolved by sodium dodecyl sulfate polyacrylamide gel electrophoresis due to their different elution from the reversed-phase support. The isolated proteins were amenable to analysis by N terminal sequencing, enzymic digestion and mass spectrometry of their proteolytic fragments. Chromatographic elution of proteins from the reversed-phase mini cartridge was apparently independent of the specific loading mode employed, i.e., loading by conventional loop injection or by electrotransfer. PMID- 7498137 TI - From image processing to classification: I. Modeling disturbances of isoelectric focusing patterns. AB - In order to optimize the conditions for evaluation of isoelectric focusing (IEF) patterns by digital image processing, the sources of error in determination of the pI values were analyzed together with the influence of a varying background. The effects of band distortions, in the spectra of the individual lanes, were examined. In order to minimize the effect of these distortions, optimal conditions for handling IEF patterns by digital image processing were elucidated. The systematic part of the global deformation on the gels was investigated and an algorithm was developed by which it was possible to correct for a part of the individual distortions. The effects of various corrections for lane distortions were illustrated by classification, using different types of discriminant analysis. Finally the background disturbances were examined, and described by a mathematical model. PMID- 7498138 TI - From image processing to classification: II. Classification of electrophoretic patterns using self-organizing feature maps and feed-forward neural networks. AB - In a recent study, isoelectric focusing patterns were classified with a neural network using the back-propagation algorithm [1]. In order to further study the classification process and to generalize the presentation of electrophoretic patterns, Kohonen's self-organizing feature maps [2] were applied in this study. Although these feature maps are very efficient in many pattern recognition tasks, our data proved to be too complex for classification with an unsupervised system. Therefore, a second supervised network on top of the feature map was necessary. As in [3], a feed-forward network trained by the back-propagation algorithm was used. The final system allows us to correctly classify 90% of all wheat varieties. Moreover, the system proved to be reliable, reasonable in training time and shows the same accuracy in different experimental setups. PMID- 7498139 TI - Imaging electrophoretic gels with a scanning beam laser macroscope. AB - A scanning beam laser macroscope has been developed which scans an area of 7.5 x 7.5 cm in 5 s. This new imaging system is examined as a potential tool for scanning electrophoretic gels. A specially-designed telecentric, f* theta laser scan lens is used in the instrument to achieve a linear scan and a flat focal plane. The laser scan lens focuses the incoming beam from a laser to a 10 microns spot inside the gel. A raster scan is performed across the gel and the signal is detected with a photomultiplier, forming a 512 x 512 digital image stored as a computer file. Silver-stained protein polyacrylamide gels have been imaged in transmission and double-transmission, while DNA agarose gels (stained with ethidium bromide) have been imaged in fluorescence with better than 25 pg sensitivity. The macroscope has the advantage that it is not tied to the electrophoresis system as are end-of-line scanners, and the scan is rapid, so that several gels can be scanned in a very short time. PMID- 7498140 TI - Multivariate statistical comparison of autoantibody-repertoires (western blots) by discriminant analysis. AB - A procedure for the quantification and comparison of complex (auto-) antibody repertoires of many individuals is described. It is based on multivariate statistical analyses of densitometric data of individual staining patterns, in this case Western blots. Implementation of algorithms can be based on standard software and hardware components. This procedure allows (i) the quantitative assessment of the reproducibility and reliability of any electrophoretic procedure, such as for the production of the immunological substrate, (ii) the statistical comparison of any group of staining patterns, e.g., derived from patients with autoimmune diseases or normal controls, (iii) the identification of the bands that contribute most to the differences between such groups, and (iv) the determination whether an unknown individual sample belongs to a known group. A statistical analysis of autoantibody repertoires has important possible applications: it detects spontaneous or therapeutically induced changes of repertoires, it identifies regions of interest, it supports the isolation of relevant antigens, and it will also be helpful in the diagnosis of autoimmune diseases. PMID- 7498141 TI - A silver stain protocol for proteins yielding high resolution and transparent background in sodium dodecyl sulfate-polyacrylamide gels. AB - A simple, sensitive silver staining method for sodium dodecyl sulfate polyacrylamide gel electrophoresis has been developed which yields high resolution of proteins with a transparent background. The method includes the incorporation of a water wash after a short fixation step in ethanol/ acetic acid and prior to glutaraldehyde cross-linking, which appears to be necessary for the high resolution staining of protein bands and the low background staining. PMID- 7498143 TI - Dynamics of peak dispersion in capillary zone electrophoresis including wall adsorption I. Theoretical model and results of simulation. AB - A mathematical model is described for the simulation of peak profiles in capillary zone electrophoresis taking wall adsorption into account. It is based on such physico-chemical relations as mass balance equations, sorption rate equations and appropriate boundary conditions. The numerical solution of the model is carried out, allowing the depiction of the concentration profile of the sample in the capillary as a result of the dynamics of the processes involved, which depend on a number of input parameters: rate constants, adsorption isotherms, column dimensions, field strength etc. Four cases are discussed in detail, namely those with either slow or fast adsorption kinetics and linear or nonlinear isotherms. PMID- 7498142 TI - Horizontal gel electrophoresis with sample volumes up to 1.5 mL, using a discontinuous buffer system and automated apparatus. AB - Horizontal gel electrophoresis has previously suffered from lack of an instrumental design that would allow for application of large sample volumes. A recently introduced commercial apparatus (HPGE-1000, LabIntelligence) remedies that problem in application to gel electrophoresis with intermittent scanning of fluorescence by using the concentrator module of that apparatus as a stacking gel reservoir. By using this technique and a 3 mL stacking gel, protein samples of up to 1.5 mL yield bands in the horizontal resolving gel that are independent of sample volume in their width, area and migration rate. A remaining procedural problem relates to the apparent nonsimultaneity of dye and protein entrance into the resolving gel, which necessitates peak characterization by absolute rather than relative mobilities. PMID- 7498144 TI - Highly sensitive chiral analysis in on-line combined chiral and achiral media by capillary zone electrophoresis. AB - A new approach is described for highly sensitive chiral analyses by capillary zone electrophoresis, based on using an on-line combination of two capillaries filled with either chiral selective or achiral background electrolytes (BGE). Thus, the BGEs are selected in such a way that the first capillary provides optimum chiral selectivity and the second one optimum detection sensitivity. Direct chiral separations of enantiomers of mandelic, m-methoxymandelic, 3 phenyllactic and 3-indolelactic acids served as a model example for testing the approach proposed. The analyses were performed in a BGE containing acetate as a coion and L-OH-proline or aspartame as a chiral selector. For high sensitive analyses, an arrangement containing on-line combined chiral and achiral media were tested in one or two capillaries coupled via a bifurcation block. A detection limit as low as 10(-18) moles was reached in the column-coupling system when the direct chiral separation was performed in the first capillary, filled with 20 mM acetate buffer, pH 4.4, containing 8 mM Cu (II) and 16 mM aspartame (L aspartyl-L-phenylalanine methylester) and separated enantiomers were detected in the second capillary, filled with 20 mM acetate buffer, pH 3.1. The principle described is of general use in cases where the separation and detection of analytes in question require mutually different BGEs to reach the optimum selectivity and sensitivity, respectively. PMID- 7498145 TI - Application of dual internal standards for precise sizing of polymerase chain reaction products using capillary electrophoresis. AB - Capillary electrophoresis (CE) is an analytical technique which provides rapid, high resolution analysis of amplified DNA fragments produced by the polymerase chain reaction (PCR). In this study, two internal standards are used as size markers to bracket und precisely size PCR products. The technique is applied to typing PCR products from the short tandem repeat locus HUMTH01. HUMTH01 consists of five to seven major alleles in the size range of 179-203 bp, with each allele four bp apart. Using this genetic marker, a population containing 97 individuals was examined with both polyacrylamide gel electrophoresis and CE. Identical genotypes were obtained with both techniques demonstrating the reliability of CE in DNA typing applications. The DNA analysis took place in sets of 10 with a calibration of the CE being performed between each set of samples. For the 97 samples examined, the pooled standard deviation was 0.3 bp. The observed genotype frequencies determined from the sample set did not deviate significantly from Hardy-Weinberg expectations. From these CE results, we conclude that HUMTH01 PCR products can be accurately and precisely sized by capillary electrophoresis using the method described. PMID- 7498148 TI - Capillary electrophoresis of human serum proteins and apolipoproteins. AB - Capillary electrophoresis (CE) analysis of human serum proteins and apolipoproteins is described and compared with sodium dodecyl sulfate polyacrylamide gel electrophoresis and cellulose acetate membrane electrophoresis. Under optimized CE conditions apolipoproteins I could be determined directly in the serum and quantitated with a linear correlation between peak area and concentration up to a concentration of 100 mg/dL. PMID- 7498146 TI - Capillary electrophoresis of rat liver microsomes in polymer solutions. AB - Rat liver microsome components were separated by capillary zone electrophoresis in buffers containing substituted agarose, agarose crosslinked polyacrylamide, polyacrylamide, polyethylene glycol and dextran of various molecular weights. The best resolution of the components was obtained with polymers of 10-21 nm geometric mean radius. Both the crude and the purified preparations of microsomes exhibit a single major peak. In a Tris-borate-EDTA (TBE) buffer, containing polyacrylamide of 5 x 10(6) molecular weight, it has a retardation coefficient, KR, of 0.77 +/- 0.02. Translation of the KR value to geometric mean radii, R, on the basis of the standard curve applicable to polymer solutions, KR vs. R, with polystyrene carboxylate size standards in both dextran and polyacrylamide solution allows one to estimate a value of R as 13-16 nm for the major microsome component. The value is smaller than expected from electron microscopic measurements (100-250 nm), possibly due to the chemical and geometric differences between microsomes and the polystyrene particles used as size standards. The crude preparation also contains a minor component which is smaller and less charged than the major component. Another component, apparently very much larger than the major one, is seen in TBE buffer but not in a potassium-N-(2 hydroxyethyl)piperazine-2'-(2-ethanesulfonic acid) buffer and is therefore thought to be an artifact of interaction with borate. After a short incubation under conditions promoting delayed microsome fusion, i.e. in presence of GTP and Mg++ and in the absence of polyethylene glycol, the electrophoretic pattern changes dramatically: it now exhibits five unretarded, highly mobile and, therefore, presumably large components in addition to the two original retarded components of the microsome and a less highly charged species similar in KR to the smaller of the original two components. PMID- 7498147 TI - Capillary and slab gel electrophoresis profiling of oligosaccharides. AB - Capillary electrophoresis with laser-induced fluorescent detection, a one dimensional version of the well-established planar analytical method of polyacrylamide gel electrophoresis, has been proven to be a powerful new microanalytical method for profiling complex carbohydrates. In this paper a comparison is presented between the planar high concentration polyacrylamide gel electrophoresis method and capillary electrophoresis of different carbohydrates with respect to performance and efficiency. N-Linked oligosaccharides were released from several glycoproteins, including fetuin, human immunodeficiency virus (HIV) envelope recombinant glycoprotein (GP-120), alpha 1-acid glycoprotein and ribonuclease B, using recombinant peptide-N-glycosidase F (PNGase F). Both separation methods involve labeling of the released carbohydrates at the reducing end with the fluorescent dye, disodium 8-amino-1,3,6-naphthalene trisulfonate (ANTS). Fluorophore labeling was followed by separation of the labeled oligosaccharides either by high concentration polyacrylamide gel electrophoresis or capillary electrophoresis. PMID- 7498150 TI - Two-dimensional polyacrylamide gel electrophoresis with immobilized pH gradients in the first dimension (IPG-Dalt): the state of the art and the controversy of vertical versus horizontal systems. AB - After having established the basic protocol of two-dimensional electrophoresis with immobilized pH gradients in the first dimension (IPG-Dalt) in 1988 (A. Gorg et al., Electrophoresis 1988, 9, 531-546), some critical parameters of the actual IPG-Dalt protocols as well as the results obtained with horizontal and vertical second-dimensional sodium dodecyl sulfate-electrophoresis are demonstrated and discussed. PMID- 7498149 TI - 2D Electrophoresis: From Protein Maps to Genomes. Proceedings of the International Meeting. Siena, Italy, September 5-7, 1994. PMID- 7498151 TI - Sample preparation of human tumors prior to two-dimensional electrophoresis of proteins. AB - Sample preparation procedures are critical for obtaining high quality two dimensional electrophoresis (2-DE) separations from clinical tumor materials. In this communication, we describe procedures which we have found satisfactory for preparing various tumor samples for 2-DE. By these procedures, tumor cells are enriched from serum proteins and contaminating stromal cells. Tumors can be prepared with good reproducibility to obtain high resolution using these techniques. PMID- 7498152 TI - Progress with gene-product mapping of the Mollicutes: Mycoplasma genitalium. AB - A protein map of the smallest known self-replicating organism, Mycoplasma genitalium (Class: Mollicutes), revealed a high proportion of acidic proteins. Amino acid composition was used to putatively identify, or provide unique parameters, for 50 gene products separated by two-dimensional gel electrophoresis. A further 19 proteins were subjected to peptide-mass fingerprinting using matrix-assisted laser desorption ionisation-time of flight (MALDI-TOF) mass spectrometry and 4 were subjected to N-terminal Edman degradation. The majority of M. genitalium proteins remain uncharacterised. However, the combined approach of amino acid analysis and peptide-mass fingerprinting allowed gene products to be linked to homologous genes in a variety of organisms. This has allowed proteins to be identified prior to detection of their respective genes via the M. genitalium sequencing initiative. The principle of 'hierarchical' analysis for the mass screening of proteins and the analysis of microbial genomes via their protein complement or 'proteome' is detailed. Here, characterisation of gene products depends upon the quickest and most economical technologies being employed initially, so as to determine if a large number of proteins are already present in both homologous and heterologous species databases. Initial screening, which lends itself to automation and robotics, can then be followed by more time and cost intensive procedures, when necessary. PMID- 7498153 TI - Amino acid compositions of proteins and their identities. AB - A critical overview is given on the application of amino acid composition data for the establishment of the protein's identity (amino acids composition vs. protein identity, the AAC-PI method). Several criteria are used to measure the differences between the amino acid compositions of various proteins. The AAC-PI method unambiguously identifies proteins which belong to the families with a high phylogenetic conservancy of their sequences. The identification of pure proteins can be accomplished with a relatively high level of confidence. The AAC-PI method, however, sometimes needs the support of N-terminal or internal sequencing of proteins since, alone, it cannot distinguish whether the lack of finding a candidate protein in protein data bases is because the investigated amino acid composition corresponds to an unknown protein or its processed form or because it is a sum of at least two protein components, or whether it is due to other experimental errors. The identification of a few new proteins such as "arginine rich protein", macrophage migration inhibitory factor (MIF) and the preformed neurotrophic factor present in the calf brain cytosol is also reported. PMID- 7498154 TI - Matrix-assisted laser-desorption/ionization mass spectrometric approaches for the identification of gel-separated proteins in the 5-50 pmol range. AB - The ability to identify and characterize low picomole quantities of gel-separated proteins has greatly benefited from recent advancements in mass spectrometric analysis methods, particularly peptide-mass search routines. We are investigating the use of matrix-assisted laser desorption/ionization mass spectrometry (MALDI MS)) to gain as much mass information as possible from a single gel-separated protein species. This report details results obtained from one-dimensional sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) separations, under nonreducing conditions, of known quantities of three proteins, followed by blotting to Immobilon-CD. Three methods were used to obtain MALDI-MS data from a single blotted protein band: (i) direct MALDI-MS of approximately 10% of the band, (ii) cyanogen bromide (CNBr) cleavage of another approximately 10% of the band, and (iii) enzymatic (endoproteinase Lys-C) digestion of the remaining approximately 70-80% of the band followed by MALDI-MS. At the level of 50 pmol of protein loaded onto the gel, data was obtained from all three approaches. At levels down to 5 pmol of protein loaded onto the gel, MALDI-MS data was obtained from the latter two methods, CNBr and Lys-C digestions, but not direct MALDI-MS. Sufficient peptide masses were obtained from the 5 pmol loads to identify two of the three test proteins using four mass search programs. Only limited peptide mass data was obtained from fetuin, a sialylated glycoprotein with six disulfides and no methionines, but it was identified. PMID- 7498155 TI - Sequence analysis of wheat grain allergens separated by two-dimensional electrophoresis with immobilized pH gradients. AB - Micropreparative two-dimensional (2-D) gel electrophoresis with immobilized pH gradients (4-8) in the first dimension (IPG-DALT) was optimized for the separation of salt-soluble wheat grain proteins, associated with bakers' asthma disease. The resolved polypeptides were electroblotted onto a polyvinylidene difluoride (PVDF) membrane and incubated with the pooled sera from four asthmatic bakers. Bound IgE was demonstrated by alkaline phosphatase conjugated anti-human IgE. Major IgE binding was detected in the 27 kDa, 37 kDa and, to a lesser extent, in the 14-18 kDa area of the 2-D immunoblots, respectively. Since the main purpose of our study was to determine the N-terminal amino acid sequences of the major wheat grain allergens, N-terminal sequencing was performed for six out of a total of eleven major allergens located in the 27 kDa area, for one out of two 37 kDa allergens, and for two out of four 14-18 kDa allergens. Our results revealed that two of the 27 kDa polypeptides are clearly related to several Acyl CoA oxidase variants of barley and rice, whereas no significant homologies were found for the remaining four 27 kDa allergens analyzed. The N-terminus of the 37 kDa allergen appeared to be blocked so that no sequence information was obtained, while the two 14-18 kDa allergens analyzed were identified as members of the wheat alpha-amylase-inhibitor family. PMID- 7498156 TI - Nonreducing two-dimensional polyacrylamide gel electrophoretic analysis of human colonic proteins. AB - Immunochemical detection of proteins with antigenic determinants that are dependent on the native spatial conformation of the protein can often pose problems with conventional two-dimensional polyacrylamide gel electrophoresis (2 DE). For example, many antigenic determinants are readily destroyed by reducing agents and/or urea, reagents which are critical components of many of the conventional isoelectric focusing and immobilized-pH-gradient (IPG) protocols used in the first electrophoretic dimension. Here we describe the use of commercially available precast 2-DE gels for performing nonreducing/non-urea 2-DE of proteins extracted from the human colon cancer cell line LIM 1215 with 0.3% Triton X-100 that permit the identification of antigens with conformational determinants by immunoblot analysis. Previous, related studies demonstrated the usefulness of peptide-mass fingerprinting for identifying 2-DE resolved proteins. Here we show how partial protein sequence data obtained by rapid peptide mapping, using capillary column liquid chromatography directly coupled with electrospray ionization tandem mass spectrometric methodologies, enhances the usefulness of this approach for identifying incompletely resolved proteins. The nonreducing 2 DE gel images reported in this study, along with our master 2-DE gel protein database for both normal human colonic crypts and several colon-cancer-derived cell lines, and information regarding microtechniques employed in this laboratory for obtaining structural data on 2-DE resolved proteins can be accessed over the Internet using World Wide Web (URL address: http:@www.ludwig.edu.au). PMID- 7498157 TI - Inside SWISS-2DPAGE database. AB - Several two-dimensional polyacrylamide gel electrophoresis (2-D PAGE) databases have been established and updated for more than 15 years. Only recently have developments of computer networks and high-speed transfer protocols provided the required tools for sharing comprehensive and hypermedia 2-D PAGE databases. This publication describes the SWISS-2DPAGE database structure. Proteins present in samples of human tissue, cells, cell lines and body fluids are assembled and described in an accessible uniform format. SWISS-2DPAGE can be freely accessed through the World-Wide Web (WWW) network on the ExPASy molecular biology server. PMID- 7498158 TI - Human blood platelet protein map established by two-dimensional polyacrylamide gel electrophoresis. AB - Two-dimensional (2-D) maps of cytosol and enriched-membrane platelet proteins has allowed the identification of more than 25 spots by three different methods: matching of the platelet gels with other 2-D reference maps, immunoblotting with chemiluminescence detection, and N-terminal sequencing. Different G protein (guanosine triphosphate-binding protein) subunits, cytoskeletal proteins, and proteins common to the human liver, red blood cells and plasma were identified. The two platelet protein maps presented here contribute to the project of identification of human cell and body fluid proteins. They may serve as working tools since platelets are popular models for the study of central nervous system neurotransmitter systems and stimulus-response coupling mechanisms. PMID- 7498159 TI - The major protein expression profile and two-dimensional protein database of human heart. AB - The construction of a two-dimensional protein database of the human heart is presented. The database contains information on about 300 abundant proteins of human myocardial tissue, including approximately 40 proteins that were identified by different methods. Each protein was characterized according to several parameters, including molecular weight, isoelectric point, name, partial sequence, subcellular localization, and genetic as well as embryonic changes. PMID- 7498160 TI - A Saccharomyces cerevisiae Internet protein resource now available. AB - The QUEST Protein Database Center is now making available two Saccharomyces cerevisiae protein databases via the Internet. The yeast electrophoretic protein database (YEPD) is a database of approximately one hundred protein identifications on two-dimensional gels. The yeast protein database (YPD) is a database of gene names and properties of over 3500 yeast proteins of known sequence. These databases can be accessed via a World-Wide Web (WWW) server (URL http:@siva.cshl.org). YPD is available via public ftp (isis.cshl.org) as well, in a spreadsheet format, and in ASCII format. When accessed via WWW, both of these databases have hypertext links to other biological data, such as the SWISS-PROT protein sequence database and the Saccharomyces Genome Database (SacchDB), and to each other. PMID- 7498161 TI - The protein disease database. PMID- 7498162 TI - Two-dimensional protein electrophoresis and multiple hypothesis testing to detect potential serum protein biomarkers in children with fetal alcohol syndrome. AB - Fetal alcohol syndrome (FAS) surveillance and intervention efforts are hampered by the lack of a specific biochemical test for diagnosis of the syndrome. Based on the hypothesis that abnormalities in growth and development (key features of FAS) involve altered protein metabolism, we analyzed serum proteins by two dimensional gel electrophoresis and image analysis to search for potential protein biomarkers of FAS. Serum samples from 12 participants in whom FAS had been diagnosed and 8 sex- and age-matched participants whose mothers did not consume alcohol were analyzed in duplicate to determine whether the integrated intensities of matched proteins are significantly altered in children with FAS. Multiple hypothesis testing on 34 of the gels consisting of more than 1700 spots per gel revealed 21 proteins that we classified as potential protein biomarkers of FAS on the basis of significant t-test differences at p < 0.02. We classified 8 of the proteins as candidate biomarkers on the basis of significant concentration differences between case and control subjects at p < 0.01. One of the proteins is clearly an isoform of retinol binding protein; two appear in the area of the gel where alcohol dehydrogenase is expected to appear; one appears to be an isoform of alpha-1-antitrypsin; three appear to be isoforms of the beta chain of haptoglobin; three may be forms of immunoglobulin light chains; and several others have not been associated with known proteins. No single protein differentiated all case subjects from control subjects, but stepwise canonical discriminant analyses revealed four groups of spots that distinguished between FAS case and control subjects with no misclassifications.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7498163 TI - Phenotyping of apolipoprotein E using immobilized pH gradient gels for one dimensional and two-dimensional separations. AB - Apolipoprotein E (apo E) is a normal component of several classes of plasma lipoproteins. Apo E phenotypes are closely related to total cholesterol, low density lipoprotein (LDL)-cholesterol and apo B concentration. The apo E 2/2 phenotype is related to the type III hyperlipoproteinemia due to the defective binding of apo E-2 to the hepatic receptors. The apo E 4/4 phenotype has been reported to be present in most elderly people suffering from the Alzheimer disease, and is associated with increased risk of coronary heart disease and Creutzfeld-Jakob disease. Therefore, apo E phenotyping is essential. The method described here uses a precast immobilized pH gradient, avoids time-consuming separation of lipoproteins from plasma, needs no pretreatment with neuraminidase and involves highly sensitive enhanced chemiluminescence for visualization. Therefore it has many advantages over previously published methods. PMID- 7498164 TI - Rapid detection of the main human plasma glycoproteins by two-dimensional polyacrylamide gel electrophoresis lectin affinoblotting. AB - Glycoprotein modifications in the glycan moiety can occur in diseases such as cancers, inflammatory processes and alcoholism. We combined high-resolution two dimensional polyacrylamide gel electrophoresis (2-D PAGE) with lectin affinoblotting in order to establish the normal human plasma glycoprotein map. Human plasma proteins were separated by mini 2-D PAGE (7 x 9 cm), transferred onto polyvinylidene difluoride membranes and incubated with biotinylated lectins. We focused our study on lectins binding sialic acid and galactose residues. Known plasma glycoproteins such as alpha 1-antitrypsin, alpha 1-antichymotrypsin, alpha 2-HS glycoprotein, alpha 1-acid glycoprotein, haptoglobin beta-chain and transferrin were easily detected in ng amounts. This protocol was adequate to establish a normal plasma glycoprotein map and will allow the study of glycoproteins in diseases. PMID- 7498165 TI - Two-dimensional electrophoresis of human serum proteins modified by ampicillin during therapeutic treatment. AB - Two-dimensional electrophoretograms of serum proteins from ampicillin-treated patients were analyzed-by immunoblotting with an antiserum specific for penicilloyl groups. As expected, human serum albumin (HSA) was the main ampicilloylated serum component. Transferrin main form II was found to be the second most important component as regards immunoblotting intensity. Immunoreactive spots were present on the acidic side of the transferrin isoelectric series, suggesting a modification mechanism similar to that observed in HSA, i.e., acylation of basic amino acid residues. Several additional ampicilloylated spots were detected but could not be assigned. Their electrophoretic parameters were determined using internal standards. This is the first description of serum proteins other than HSA being modified by ampicillin in the course of routine therapeutic treatment. PMID- 7498166 TI - Two-dimensional electrophoresis for the study of blood/serum proteins of the otter, an endangered species. AB - Studies on serum/blood samples from European otters (Lutra lutra), a species under protection in most countries, were undertaken. Building of a two dimensional electrophoresis (2-DE) protein identification map was started, identifying serum proteins by pattern comparison with cat or human serum protein maps and by immunoblotting using cross-reacting antibodies against the respective human serum proteins. The capture of endangered species is very restricted, thus making fresh samples almost unavailable; most investigations therefore had to be performed on samples collected from animals found dead. Patterns obtained from specimens of this source were compared to the established pattern of serum proteins and of blood or liver cells. Some of the major differences observed can be explained by contamination with cellular proteins and/or the onset of lytic processes generating breakdown products, as shown by preliminary in vitro experiments. Better knowledge of "normal" serum protein patterns and interfering products will facilitate interpretation of data collected from samples with a less defined history. PMID- 7498167 TI - Two-dimensional gel electrophoresis of nasal and bronchoalveolar lavage fluids after occupational exposure. AB - Human nasal lavage fluids (NLFs), and bronchoalveolar lavage fluids (BALFs) were analyzed with two-dimensional gel electrophoresis (2-DE) and the protein patterns were evaluated with a computerized imaging 2-DE system. With silver staining about 1000 spots were detected in 10 microgram samples of NLF or BALF. Both BALF and NLF 2-DE patterns showed similarities to a reference plasma pattern and about 25 plasma proteins were identified in NLF as well as in BALF. Comparison showed that the levels of albumin and transferrin appeared to be slightly higher in BALF than NLF, while the levels of IgA, IgG and haptoglobin beta were higher in NLF than in BALF. In contrast to BALF and blood plasma, NLF contained large amounts of a cluster of acidic proteins (pI 4.5-5.5) with molecular masses of 15-30 kDa. Distinct alterations in the NLF 2-DE patterns were found in a worker who developed an asthmatic condition with bronchial hyperreactivity after exposure to organic acid anhydrides. After exposure, 14 protein spots were increased and one decreased by a factor of > 3 as compared to the levels before exposure and compared to healthy individuals. This is the first study indicating that 2-DE of NLF may be used to investigate early changes in airway protein patterns induced by occupational exposure to irritating chemicals. PMID- 7498168 TI - Effects of oxidative stress and Ca2+ agonists on molecular chaperones in human umbilical vein endothelial cells. AB - Endothelial cell dysfunction is a key factor in oxidative stress-related pathology. Disruption of Ca2+ homeostasis is thought to be responsible for much of the endothelial cell dysfunction in oxidative stress. The expression of molecular chaperones (MC), which stabilize protein structures in normal and in stress conditions, reflects the Ca(2+)-dependent and -independent stress effects in the different cell compartments. By two-dimensional (2-D) gel electrophoresis combined with immunoblotting or microsequencing, we have identified 12 major MC in human umbilical vein endothelial cells (HUVEC): (i) the endoplasmic reticulum located MC GRP78, GRP94, protein disulfide isomerase, and calreticulin; (ii) the mitochondrial MC HSP65 and GRP75; and (iii) the cytosolic/nuclear MC HSP27, HSC70, HSP70, HSP90, cyclophilin, and ubiquitin. To differentiate oxidative stress- and Ca(2+)-mediated effects, HUVEC were exposed to 1) xanthine oxidase plus hypoxanthine to generate oxidative stress, 2) ionomycin plus ethylene glycol bis(beta-aminoethylether)-N,N,N', N'-tetraacetic acid (EGTA) to deplete intracellular Ca2+ stores, or 3) thrombin to increase cytosolic Ca2+. De novo protein synthesis after exposure was quantified by the incorporation of [35S]methionine. Image processing with the MELANIE system was used to create and compare the 2-D maps of [35S]methionine-labeled proteins under conditions 1)-3) with those of the controls. In a total of 24 2-D gels, 9 different MC were detected in at least 5 out 6 experimental replicates and were subjected to numeric analysis. The statistics showed a > 10% increase in GRP78 (p < 0.05), HSP27, cyclophilin, and ubiquitin after oxidative stress.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7498169 TI - Analysis of proteins from human breast epithelial cells using two-dimensional gel electrophoresis. AB - The human breast is a highly specialized, complex tissue comprised of a heterogeneous population of cells with varying functions. Interactions between the different cell types, changes in their relative abundance, state of differentiation and function in response to stimuli, as well as the alterations that lead to the aberrant growth associated with malignancy are poorly understood. Two-dimensional gel electrophoresis is being used to compare the proteins found in different breast cells in order to identify the gene products that are common or specific to particular cell types so as to provide markers that will be useful in studies of normal breast cell differentiation and the dedifferentiation or blocked differentiation characteristic of cancer. Protein patterns have been obtained from cells prepared for electrophoresis immediately after isolation from human milk, from cells cultured for fewer than ten passages after isolation from healthy breast tissue removed during reduction mammoplasty, and from cells maintained in long-term tissue culture after isolation from the pleural effusions of patients with breast carcinomas. Differential expression of cytokeratins 8, 18, and 19, shown previously to be predominantly expressed by epithelial cells in the luminal layer of breast tissue, was observed among the cells analyzed. Other non-cytokeratin proteins were also found to be differentially expressed in subsets of both the normal and tumor cells. A composite human breast cell protein pattern was created which includes all the commonly and specifically expressed proteins found in this study. This pattern will be the basis for continuing studies of proteins in the human breast. PMID- 7498171 TI - Analysis of growth factor stimulated nucleocytoplasmic protein transport using two-dimensional gel electrophoresis. AB - The rapid activation of gene expression by growth factors frequently involves the nucleocytoplasmic redistribution of proteins which regulate the transcription of discrete sets of genes. In particular, it has become apparent that protein kinases and transcription factors including protein kinase A, mitogen activated protein kinase, protein kinase C, NF-kappa B and Stat91, can rapidly migrate from the cytosol to the nucleus in response to a wide variety of extracellular stimuli. Furthermore, it is assumed that these proteins represent important components of the signal transduction pathways which operate between the plasma membrane and the nucleus. Here two-dimensional polyacrylamide gel electrophoresis (2-D PAGE) has been used to detect proteins which undergo rapid growth factor stimulated nucleocytoplasmic redistribution. Epidermal growth factor and insulin influenced the association of 15 individual protein spots with the nucleus. Estimates of the abundance of these proteins revealed that they represented approximately 1/1000 to 1/5000 of total nuclear protein. To permit identification by microsequencing, five of the protein spots have been recovered from multiple 2 D PAGE gels and concentrated by one-dimensional elution-concentration gel electrophoresis. The data obtained demonstrate that 2-D PAGE can be used to: (i) analyse putative regulators of growth factor activated gene transcription, and (ii) purify sufficient amounts of these proteins to facilitate identification by microsequence analysis. PMID- 7498170 TI - Spermatocytes and round spermatids of rat testis: protein patterns. AB - Spermatogenesis is a process in the testis that involves meiotic cell division and spermiogenesis. The mechanisms of regulation and its associated proteins are mostly unknown. This publication shows the two-dimensional (2-D) gel electrophoresis protein map obtained from rat testis using nonlinear 3.5-10 immobilized pH gradients for the first-dimensional separation. Eighteen proteins were successfully identified in the SWISS-PROT protein database using amino acid analysis of proteins recovered from polyvinylidene difluoride (PVDF) membranes and verified for one of them by comparison with Anderson's rat liver reference map. Fourteen new polypeptides were identified and four were previously known. Two of these new proteins were closely related to the spermatogenetic process. T complex protein 1 is expressed in large amounts in germ cells. Androgen-dependent sperm-coating glycoprotein is secreted by epididymal cells. In order to detect changes in protein expression during meiosis and spermiogenesis, spermatocytes and round spermatid cell populations were purified by centrifugal elutriation and compared. In this way several proteins not found in the spermatocyte 2-D images could be high-lighted. The sperm-coating glycoprotein was thus shown to be present in large amounts in round spermatids. PMID- 7498172 TI - Long-term induction of an aldose reductase protein by basic fibroblast growth factor in rat astrocytes in vitro. AB - Basic fibroblast growth factor (bFGF) is known to elicit various developmental like effects on astrocytes in vitro, but these effects were studied mainly over short-term periods. In this work we asked the question whether bFGF could induce long-term effects on rat astrocytes in culture. This factor was found to induce only a transient mitogenic effect lasting less than 48 h, even when the treatment was carried on for 4 days. By contrast, it induced long-term effects on the rate of synthesis of several proteins as seen by two-dimensional polyacrylamide gel electrophoresis after labeling the cells with [35S]methionine. The most upregulated protein was extracted from preparative gels of soluble extracts of cultured bFGF-treated astrocytes and of normal brain. It was characterized by internal amino acid microsequencing. Two tryptic digest peptides had N-terminal sequences similar to rat lens aldose reductase. This protein was also expressed in oligodendroglial and neuronal cells in culture, but it was not upregulated by bFGF. Aldose reductase is thought to be involved in a minor pathway of glucose metabolism and in diabetic complications. Its long-term regulation by bFGF will possibly help in the understanding of its actual physiological role. PMID- 7498174 TI - Programmed cell death in sympathetic neurons: a study by two-dimensional polyacrylamide gel electrophoresis using computer image analysis. AB - The technique of two-dimensional polyacrylamide gel electrophoresis (2-D PAGE) coupled with computer image analysis was used in this study to examine changes in protein expression occurring during the onset of programmed cell death (PCD) in rat sympathetic neurons following withdrawal of nerve growth factor (NGF). Sympathetic neurons from superior cervical ganglia of postnatal day-one Wistar rats were cultured in the presence of NGF for 24 h and then either maintained in the presence of NGF or deprived of NGF for a period of 8 h. To label the proteins being synthesised, neurons were cultured in the presence of L-[35S]methionine for a further 2 h under the same conditions but with 3% of the normal methionine concentration. Neuronal proteins were then analysed by 2-D PAGE using immobilised pH gradient (IPG) gel strips in the first dimension. For the second dimension a custom-built electrophoresis system capable of running multiple sodium dodecyl sulfate (SDS)-PAGE slab gels in a vertical configuration, with good temperature control (+/- 0.7 degrees C) was used and is described in this paper. Proteins resolved on the dried gels were visualised using storage phosphor technology and the digitised images subjected to rigorous analysis using the QUEST II software system. Seventeen proteins whose relative synthesis decreased and four proteins that increased upon NGF withdrawal were located and are documented. PMID- 7498173 TI - Repetitive treatment with serotonin modifies protein synthesis and protein phosphorylation in the central nervous system of Hirudo medicinalis. AB - Serotonin (5HT) is the neurotransmitter involved in some forms of short-term memory in the leech. Behavioral experiments have demonstrated that long-term memory requires new protein synthesis. With the aim of studying the molecular mechanism underlying memory processes in the leech, we have analyzed the effect of 5HT on protein synthesis and protein phosphorylation. Segmental ganglia of the leech central nervous system have been labeled, proteins have been separated by two-dimensional-electrophoresis and labeled proteins detected by autoradiography. Our findings indicate that repetitive treatment with 5HT produces either the persistence of phosphorylation or changes in protein synthesis in several proteins. PMID- 7498175 TI - Two-dimensional electrophoresis of intracellular and secreted protein synthesized by fetal bovine chondrocytes in high-density culture. AB - In order to study the mechanisms involved in the differentiation/dedifferentiation of chondrocytes, fetal bovine chondrocytes in high-density cultures were treated with retinoic acid, an agent known to modify the chondrocyte phenotype (10 mumol/L between day 2 to day 5 of culture). The synthesis of intracellular and secreted proteins was studied by two-dimensional electrophoresis in cell lysates and culture media after labeling with [35S]methionine for the last 14 h of culture. The proteins expressed in control and retinoic acid-treated cells were identified by microsequencing after "in-gel" tryptic digestion of the spot or by immunodetection with specific antibodies after two-dimensional gel blotting. Intracellular protein modifications included one of 56.9 kDa and with an isoelectric point (pI) of 5.8 whose synthesis was previously reported to be up-regulated by 75%. Microsequencing of two internal peptides did not reveal a known protein. Changes to the chondrocyte phenotype were also recorded in the culture medium, as a decrease in type II collagen synthesis and expression of the small proteoglycan, decorin. Several new spots were also observed after treatment with retinoic acid, including a large, diffuse spot, not yet characterized, with a mean molecular mass of 39 kDa and a pI of 4.5 5.0. Under our experimental conditions, retinoic acid induces morphological changes of the chondrocytes and dramatic changes in the synthesis of several intracellular and secreted proteins that predate the synthesis of collagen type I (the classical marker of chondrocyte dedifferentiation). PMID- 7498176 TI - Effects of cyclosporine A on the rat liver and kidney protein pattern, and the influence of vitamin E and C coadministration. AB - The effects of cyclosporine A (CsA), a potent immunosuppressive drug, were examined in rat liver and kidney samples using two-dimensional electrophoretic protein analysis. Of a total of 370 liver and 336 kidney spots analyzed, 8% (29 spots) and 6% (19 spots), respectively, showed a significant drug-induced change (p < 0.01), which was predominantly reflected in increased protein abundance (62% and 74% of the changes, respectively). Of the 48 proteins changed in either organ, 14 were most probably common to both tissues and one of these was significantly increased in both the liver and the kidney. Most of the other 13 showed similar trends (either increases or decreases) in both organs. However, the most striking drug effect seen in this study concerned an unidentified protein present only in the kidney, which completely disappeared upon CsA treatment. It was also investigated whether the drug-induced changes could be prevented by the coadministration of the radical scavengers vitamin E and C with CsA. Spots changed by the administration of the drug were classified according to three different categories, based on their response profiles in rats treated with CsA in combination with the vitamins: (i) spots which were changed by CsA as well as by CsA in combination with the vitamins (12 liver and 4 kidney spots), (ii) spots which were changed by CsA and showed an additional increase of this change by CsA plus the vitamins (no liver and 4 kidney spots), and (iii) spots which were changed by CsA but not by CsA in combination with the vitamins (8 liver and 6 kidney spots). These results showed that in both organs the vitamins were able to prevent around 30% of the effects caused by CsA, and that two-dimensional gel electrophoresis is an excellent tool to demonstrate such drug interactions at the molecular level. PMID- 7498178 TI - Characterizing allelic proteins for genome mapping in maize. AB - Mapping cDNA probes in order to construct genetic linkage maps is becoming a widespread strategy for genome analysis and gene isolation, particularly in cultivated plant species. Nevertheless, almost all cDNAs reveal two or more unlinked loci, making it difficult to identify the gene(s) actually expressed. In a highly polymorphic species, such as maize, two-dimensional polyacrylamide gel electrophoresis (2-D PAGE) of proteins may circumvent this limitation. With the analysis of various segregating populations, we previously showed that the apparent position shifts of proteins on the 2-D gels are monogenic and codominant. In this paper we compared allelic proteins on the basis of their high performance liquid chromatography (HPLC) profile and partial amino acid sequences. In a sample of 20 position shifts, the allelic proteins appeared to be similar in all but one case, strongly suggesting that polymorphism of structural genes is involved. Thus 2-D PAGE could prove to be a useful tool for genome mapping: when a cDNA probe detects several loci, a position shift of the encoded protein will allow the identification of the gene translated in the organ considered. PMID- 7498177 TI - Separation of the two most closely related isoenzymes of alkaline phosphatase by two-dimensional electrophoresis. AB - Seminoma is the most frequent testicular germ cell tumor. While effective curative treatment of the disease is available today, there is to date no tumor marker suited for the diagnosis and follow-up. Several authors have suggested that the germ-cell-specific isoenzyme of alkaline phosphatase (GCAP) might be valuable for this purpose. GCAP shows 98% sequence homology with the placental isoenzyme of alkaline phosphatase (PLAP). Both display a high degree of phenotypic heterogeneity. Until now all attempts to raise an antibody reacting specifically with GCAP have failed. Consequently there is no immunological assay that allows the measurement of GCAP in the presence of PLAP. Two-dimensional electrophoresis with a sigmoid immobilized pH-gradient of 3-10 for the first dimension makes it possible to differentiate clearly between these two closely related isoenzymes. Additionally, it resolves their many phenotypic variants. This is of special interest, since malignant transformation affects the glycosylation patterns of many glycoproteins. For the detection of GCAP and PLAP in two-dimensional electrophoresis it is essential to purify the raw tissue extracts thoroughly. A chromatographic method suited for this purpose is presented. PMID- 7498180 TI - Two-dimensional electrophoresis of thylakoid protein patterns in two wheat cultivars with different sensitivity to sulfur dioxide. AB - Plants of two wheat cultivars (Triticum aestivum L., cultivars Mec and Chiarano), differing in sensitivity towards sulfur dioxide, were fumigated with 96 nL L-1 of SO2 for four months. Thylakoid membranes were subjected to two-dimensional electrophoresis in order to detect possible changes in the protein patterns. We found the greatest alterations following treatment in the polypeptide profiles of the most sensitive cultivar, Mec, even if some significant changes were also detected in the plants of the cultivar Chiarano. PMID- 7498179 TI - Distance indices in a comparison between the A, D, I and R genomes of the Triticeae tribe. AB - Two-dimensional gel electrophoresis of shoot proteins was used to study the relationships between Triticum, Secale, and Hordeum. A high level of polymorphism was found among the 1275 spots scored: only 198 spots were found common to all. But, under the hypothesis that only allelic variations were observed, the mean number of alleles per locus was only two. Phenograms were built from different distance indices. All of them showed Triticum genomes A and D close to each other, Hordeum far from the Triticum cluster, and Secale at an intermediate position. A discussion on the use of various distance indices is presented. PMID- 7498181 TI - Two-dimensional electrophoresis of acidic proteins isolated from ozone-stressed Norway spruce needles (Picea abies L. Karst): separation method and image processing. AB - The effect of ozone stress (< 10,200 or 400 micrograms m-3) on the protein pattern of Picea abies L. Karst needles was studied by fumigating potted grafts from mature trees. Clonal trees of Norway spruce were fumigated in open-top chambers for one growing season. Proteins, soluble and membrane bound, were extracted from the needles in a four-step procedure: (i) grinding in liquid nitrogen and then in a pH 5.0 buffer with thiourea added to inhibit phenol oxidase and polyvinylpolypyrrolidone (PVP) added to bind phenolic compounds; (ii) precipitation of proteins with acetone at -20 degrees C; (iii) solubilization in a modified lysis buffer; and (iv) dialysis against the modified lysis buffer. Proteins were separated by two-dimensional electrophoresis according to O'Farrell's method. The changes in the silver-stained protein pattern were evaluated by using image processing methods. Electrophoretograms from stressed and nonstressed trees are presented. PMID- 7498182 TI - Genetic variability of carrot seed proteins analyzed by one- and two-dimensional electrophoresis with immobilized pH gradients. AB - The genetic variability of eight carrot populations representing commercially important morphological types was analyzed for seed protein polymorphism by one- and two-dimensional electrophoresis using immobilized pH gradients (IPG) in order to assess the potential of both techniques in carrot hybrid breeding as well as genetic quality testing of carrot trade seeds. The digitized silver-stained one- and two-dimensional gels of the urea/detergent-soluble seed proteins were subjected to computer analysis to estimate both the inter- and intrapopulation genetic variability. The results indicated that isoelectric focusing (IEF-IPG) of seed proteins is a powerful tool for quick screening of the genetic variability present between and within carrot populations, while two-dimensional polyacrylamide gel electrophoresis (IPG-DALT) can be employed to establish detailed genetic relationships of carrot populations selected by initial IEF-IPG screening in order to use genetic distant and homogeneous populations for successful carrot hybrid breeding. However, the applicability of IEF-IPG in genetic quality testing of carrot seeds seems to be restricted to hybrid purity testing. IPG-DALT is a far more powerful tool to profile unambiguously all carrot varieties commercially used in order to test the genetic quality of trade seeds, because this technique exhibits protein spots or spot combinations exclusively found in each of the eight varieties investigated. PMID- 7498183 TI - Phosphorylation of herpes simplex virus type 1 Us11 protein is independent of viral genome expression. AB - The Us11 protein is a true late gene product of herpes simplex virus type 1 (HSV 1), whose exact function is unknown but which exhibits RNA-binding properties and which is phosphorylated on serine residues. In order to determine whether the Us11 protein is phosphorylated by cellular kinase(s) or by virally encoded kinase(s), the Us11 gene has been cloned and transiently expressed in HeLa cells. In addition, HeLa-derived cell lines have been selected for their ability to express Us11 protein constitutively. 32P-Labeling and analysis by two-dimensional electrophoresis of transiently and constitutively expressed Us11 protein demonstrated that, indeed, multiple phosphorylation of the protein occurs in absence of HSV-1 genome expression, indicating that the protein behaves as a natural substrate for cellular kinase(s). In addition, a sequence heterogeneity of the Us11 protein, due to a difference in the number of SPREPR repeats, has been characterized between different strains of HSV-1. PMID- 7498184 TI - Warning signal words: connoted strength and understandability by children, elders, and non-native English speakers. AB - Signal words, such as DANGER, WARNING and CAUTION, are commonly used in sign and product label warnings for the purpose of conveying different levels of hazard. Previous research has focused on whether people's perceptions of connoted hazard are consistent with the levels suggested by design standards and guidelines. Most investigations have used college students to evaluate the terms; other populations who may be at greater risk have not been adequately studied. One purpose of the present research was to determine whether young children, the elderly, and non-native English speakers perceive similar connoted hazard levels from the terms as undergraduates and published guidelines. A second purpose was to assess the terms' comprehensibility using various metrics such as missing values (i.e. ratings left blank) and understandability ratings. A third purpose was to develop a list of potential signal words that probably would be understandable to members of special populations. In the first experiment, 298 fourth- to eighth-grade students and 70 undergraduates rated 43 potential signal words on how careful they would be after seeing each term. The undergraduates also rated the terms on strength and understandability. In the second experiment, 98 elders and 135 non-native English speakers rated the same set of terms. The rank ordering of the words was found to be consistent across the participant groups. In general, the younger students gave higher carefulness ratings than the undergraduates. The words that the younger children and the non-native English speakers frequently left blank were given lower understandability ratings. Finally, a short list of terms was derived that 95% or 99% of the youngest students (fourth- and fifth-graders) and 80% of the non-native English speakers understood. Implications of hazard communication are discussed. PMID- 7498185 TI - Interaction of signal word and colour on warning labels: differences in perceived hazard and behavioural compliance. AB - Previous research has examined the connoted hazard of various colour and signal words separately. The purpose of the present research was to examine the interaction of signal words and colours. Two separate experiments examined the influence of colour on perceptions of hazard and on compliance to printed warnings. In experiment 1, a sample of 30 undergraduates rated the perceived hazard of signal words printed in specific hazard colours. Results indicated that the level of hazard varied as a function of the signal word and the colour in which it was presented. Of the colours used, red conveyed the highest level of perceived hazard followed by orange, black, green and blue. More importantly, it was noted that a signal word such as DEADLY connoted less hazard when printed in green than in red ink. Experiment 2 examined the effect of colour on compliance with printed warnings. Sixty-five undergraduates interacted with a pool-water test kit and a two-part adhesive. The warning on each product was factorial for colour (i.e. red, green and black). Behavioural compliance was assessed by indicating if subjects donned protective gloves as directed by the warning. The data indicated that warnings printed in red resulted in a higher proportion of compliant behaviour than green and black combined. Implications for warning design are discussed. PMID- 7498186 TI - An examination of consumer advisement warning information embedded within written instructions: implications for memory and behaviour. AB - The effects of the colour and placement of consumer advisement warning information embedded within written instructions on both memory for, and compliance with, information were examined. Specifically, undergraduate subjects were instructed to read a set of written instructions for operating a camera. Subjects read consumer advisement warning information typed in either red, blue, or black ink, which was located at either the top, middle, or bottom of the written instructions. After reading the instructions, subjects were asked to take pictures using the camera. Compliance with the information and subsequent memory for the information were assessed. Results indicated that for memory there was an interaction between colour and placement, and for behaviour there was a main effect of placement. The implications of this interaction for memory of consumer advisements is discussed. In addition, the possible role of perceived importance of the advisement information in the prediction of memory and behaviour is considered. PMID- 7498187 TI - A user-centred approach to the design and evaluation of auditory warning signals: 1. Methodology. AB - This paper presents a method for the design and evaluation of auditory warning signals modelled on an existing internationally standardized method for evaluating public information systems (ISO/DIS 7001: 1979). The procedure is essentially user-centred, capitalizing upon users' associations between sounds and their meanings. The procedure is presented in a step-by-step manner, from the initial identification of referents for which warnings might be required, through the generation of ideas for warning sounds, an appropriateness ranking test, a learning and confusion test, an urgency mapping test, a recognition test and an operational test. Practical issues are discussed with respect to each of the stages, and suggestions are made as to courses of action that might be taken if problems are encountered. Theory behind the relationship between sound and meaning is discussed with reference to the practical issues addressed. PMID- 7498188 TI - Audibility of reverse alarms under hearing protectors for normal and hearing impaired listeners. AB - The question of whether or not an individual suffering from a hearing loss is capable of hearing an auditory alarm or warning is an extremely important industrial safety issue. The ISO Standard that addresses auditory warnings for workplaces requires that any auditory alarm or warning be audible to all individuals in the workplace including those suffering from a hearing loss and/or wearing hearing protection devices (HPDs). Research was undertaken to determine how the ability to detect an alarm or warning signal changed for individuals with normal hearing and two levels of hearing loss as the levels of masking noise and alarm were manipulated. Pink noise was used as the masker and a heavy-equipment reverse alarm was used as the signal. The rating method paradigm of signal detection theory was used as the experimental procedure to separate the subjects' absolute sensitivities to the alarm from their individual criteria for deciding to respond in an affirmative manner. Results indicated that even at a fairly low signal-to-noise ratio (0 dB), subjects with a substantial hearing loss [a pure tone average (PTA) hearing level of 45-50 dBHL in both ears] were capable of hearing the reverse alarm while wearing a high-attenuation earmuff in the pink noise used in the study. PMID- 7498189 TI - Human probability matching behaviour in response to alarms of varying reliability. AB - The goals of this research were to substantiate the existence of the cry-wolf effect for alarm responses, quantifying its effect on operator performance. A total of 138 undergraduate students performed two blocks of a cognitively demanding psychomotor primary task; at the same time, they were presented with alarms of varying reliabilities (25, 50 and 75% true alarms) and urgencies (green, yellow and red visual alarms presented concurrently with low-, medium- and high-urgency auditory civilian aircraft cockpit alarms). Alarm response frequencies were observed and analysed, and t-tests and repeated-measures MANOVAs were used to assess the effects of increasing alarm reliability on alarm response frequencies, speed and accuracy. The results indicate that most subjects (about 90%) do not respond to all alarms but match their response rates to the expected probability of true alarms (probability matching). About 10% of the subjects responded in the extreme, utilizing an all-or-none strategy. Implications of these results for alarm design instruction and further research are discussed. PMID- 7498190 TI - Perceived urgency of and response time to multi-tone and frequency-modulated warning signals in broadband noise. AB - Several pulse parameters that were believed to affect the perceived urgency and response time to auditory warning signals were investigated in a factorial experiment. The independent variables included pulse format (sequential, simultaneous, and frequency-modulated pulses), pulse level (65 and 79 dBC), and inter-pulse interval (0, 150 and 300 ms). The applications environments of interest were those having steady-state broadband noise. A probability monitoring task from the Criterion Task Set was used as an operator loading task to impose additional attentional demands during the signal detection and response task. The psychophysical methods of free-modulus magnitude estimation and paired comparison were used to measure subjective perceived urgency. An objective measure of response time to the signal was also obtained. Multivariate statistical analyses indicated that response time decreased significantly as perceived urgency increased. Perceived urgency of the signal increased and response time decreased as pulse level increased. Sequential signals took longer to detect and were rated as less urgent than the other two signal types. Shorter inter-pulse interval was associated with greater perceived signal urgency. PMID- 7498191 TI - A psychophysiological evaluation of the perceived urgency of auditory warning signals. AB - One significant concern that pilots have about cockpit auditory warnings is that the signals presently used lack a sense of priority. The relationship between auditory warning sound parameters and perceived urgency is, therefore, an important topic of enquiry in aviation psychology. The present investigation examined the relationship among subjective assessments of urgency, reaction time, and brainwave activity with three auditory warning signals. Subjects performed a tracking task involving automated and manual conditions, and were presented with auditory warnings having various levels of perceived and situational urgency. Subjective assessments revealed that subjects were able to rank warnings on an urgency scale, but rankings were altered after warnings were mapped to a situational urgency scale. Reaction times differed between automated and manual tracking task conditions, and physiological data showed attentional differences in response to perceived and situational warning urgency levels. This study shows that the use of physiological measures sensitive to attention and arousal, in conjunction with behavioural and subjective measures, may lead to the design of auditory warnings that produce a sense of urgency in an operator that matches the urgency of the situation. PMID- 7498192 TI - The semantic associations of acoustic parameters commonly used in the design of auditory information and warning signals. AB - Two experiments are described in which the effects of acoustic parameters, and changes in those acoustic parameters, on the salience of 42 pre-selected adjectives are explored. In experiment 1 four sets of seven stimuli each varying only along one of the four acoustic parameters pitch, speed, inharmonicity and rhythm were scaled along the 42 adjectives. Parameter-adjective associations showing significant effects both in terms of consistency (Kendall's W') and in linear mapping (regression) numbered 7 for pitch, 3 for speed, 2 for inharmonicity and 1 for rhythm. Three of the four parameters produced significant levels of both consistency and linear mapping for the adjective 'urgent'. In the second experiment, the overall meaning of complete within-parameter stimulus sets were explored and many differences both within and between parameters and adjectives were found. The results were used to design five trend monitoring sounds for helicopter flight, three of which are described in detail in this paper. PMID- 7498193 TI - Cardiac involvement in patients with myotonic dystrophy: characteristic features of magnetic resonance imaging. AB - Structural alterations of the myocardium, such as fibrosis and fatty infiltration, were observed in post-mortem examinations in patients with myotonic dystrophy, a familial multisystem neuromuscular disease with frequent cardiac involvement. To identify structural and anatomical abnormalities of the heart, 14 patients, aged 45 +/- 14 years, belonging to seven families, suffering from myotonic dystrophy were studied. Twelve-lead ECG, high resolution signal-averaged ECG, 24 h Holter monitoring, bidimensional echocardiography and cardiac magnetic resonance (MRI) were performed in all patients. Atrioventricular and/or intraventricular conduction disturbances were present in 11 patients; no major arrhythmias were recorded by Holter monitoring. Ventricular late potentials were present in four patients, absent in eight and not assessed in two (due to left bundle branch block). Echocardiogram showed abnormal findings (left ventricular hypertrophy, mitral valve prolapse, wall motion abnormalities) in eight patients. MRI revealed various cardiac alterations in 11 cases, specifically: left ventricular hypertrophy in seven, right ventricular hypertrophy in two, right ventricular enlargement in six, area of fatty infiltration and fibrosis in the right ventricle in six and in both ventricles in three. Although no clear correlations between the extent of fibro-lipomatous infiltrations and severity of cardiac dysfunction were found, fatty infiltrations were always observed in the most severely diseased patients and were frequently associated with the presence of more advanced conduction disturbances.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7498194 TI - False diagnosis of myocardial infarction due to an extreme left sided cardiac rotation in two patients. AB - We present two patients with an extremely rare condition: abnormal cardiac levoposition. Alterations in the ECG caused by this congenital abnormality and additional chest symptoms led to the false diagnosis of previous myocardial infarction. Chest X-ray and echocardiography suggested cardiac malformation. Correct diagnosis of cardiac levoposition was established by magnetic resonance imaging. PMID- 7498195 TI - Active myocardial damage in hyperthyroidism. A concurrent mechanism of heart failure reversed by treatment. AB - A patient with hyperthyroidism, heart failure, and depressed ejection fraction, in whom myocardial damage was evidenced by In111-labelled monoclonal antimyosin antibodies, is presented. Both myocardial damage and left ventricular dysfunction disappeared after antithyroid therapy. This suggests that, in addition to volume overload, heart failure in hyperthyroidism could be also due to the presence of potentially reversible concurrent myocardial damage. PMID- 7498196 TI - Coronary interventions in Europe 1992. PMID- 7498197 TI - Arrhythmia catheter ablation: new prospects. PMID- 7498198 TI - Ablation of accessory pathways during atrial fibrillation--adapting to change. PMID- 7498200 TI - The lungs in chronic heart failure. PMID- 7498199 TI - Education from RITA. Randomized Intervention Treatment of Angina. PMID- 7498202 TI - Vein plasma endothelin-1 and cyclic GMP increase during coronary angioplasty is related to myocardial ischaemia. AB - Endothelin-1 and cyclic guanosine monophosphate (c-GMP) peripheral vein plasma levels increase during coronary angioplasty, but the reason for this increase has not been elucidated. The purpose of this study was to investigate whether these changes are related to myocardial ischaemia, or to mechanical artery injury induced during the procedure. Thirty-two patients with stable angina pectoris and a single lesion were studied. They were aged 56 +/- 8 and were undergoing balloon angioplasty. Eight arteries were totally occluded and 24 were partially occluded. Blood samples were drawn from a peripheral vein after coronary artery engagement with the guiding catheter (baseline), after the first balloon inflation, immediately after the end of the procedure, and 4 h later. In the total occlusion group endothelin-1 increased by 7% (P ns), whereas in the partial occlusion group it increased by 45% after the procedure (P < 0.001). c-GMP in the partial occlusion group increased by 41% (P < 0.001) after the procedure whereas in the total occlusion group it increased by 5% (P ns). Thus, the increase in endothelin 1 and c-GMP peripheral vein plasma levels after coronary angioplasty is related to myocardial ischaemia rather than to mechanical artery injury. PMID- 7498201 TI - The diagnostic and prognostic importance of ambulatory ST recording compared to a predischarge exercise test after an episode of unstable angina or non-Q wave myocardial infarction. AB - The diagnostic and prognostic value of ambulatory ST recordings after admission to the CCU and before discharge was compared to a symptom-limited predischarge exercise test in 170 men with unstable angina pectoris or non-Q wave myocardial infarction. ST depression in recordings before discharge identified a small group of patients (18%) out of whom 23% had a myocardial infarction within 3 months compared to 7% in those without this finding. The exercise test gave more diagnostic information, with ST depression found in 52% including 70% of those with ST depression at Holter monitoring. After 3 months, 13% of patients with ST depression at exercise test had a myocardial infarction compared to 5% in the other patients. ST depression at exercise also indicated an increased risk of myocardial and future severe angina over a longer time period. Thus ST recordings are recommended before discharge in all patients after an episode of unstable coronary artery disease as it identifies the patients with the most severe prognosis who might benefit from early revascularization. In those without ST depression at Holter a predischarge exercise test will give further information regarding the long-term risk for angina and coronary events. PMID- 7498203 TI - A late increase in free radical activity post myocardial infarction. AB - Free radicals (FR) are a highly reactive chemical species which have been implicated in the pathogenesis of reperfusion injury. Experimental models of reperfusion injury have demonstrated that FR scavengers improve myocardial salvage and thus it has been postulated that they may be of benefit in acute myocardial infarct (MI) patients treated with thrombolysis. Previous studies of FR activity post MI have focused on the immediate post reperfusion period; the present study investigates FR activity in post MI patients over a longer time span. Free radicals have a very short half-life and in clinical studies their activity is usually assessed indirectly by measuring either the level of FR reaction products such as malondialdehyde (MDA) or FR scavengers such as plasma thiols (PSH). In the presence of increased FR activity MDA levels increase and PSH levels decrease. Twenty-two acute MI patients had blood samples taken on admission, day 7 and day 21 post MI for measurement of MDA and PSH levels. On day 7 post MI a significant increase in MDA was detected P = 0.0001 (Sign test) (median change +2.1 mumol.l) and a significant decrease was detected in PSH P = 0.04 (Sign test) (median change, -38 mumol.l-1). No significant differences were detected between admission and day 21 levels of MDA and PSH. This study demonstrates the presence of enhanced FR activity at a later time point than has previously been recognised. White blood cells utilise FRs during phagocytosis and the late rise in FR activity detected in this study may reflect white blood cell removal of necrotic myocardium. PMID- 7498204 TI - Protective effect of beta-blockade on dipyridamole-induced myocardial ischaemia. Role of heart rate. AB - This study was designed to investigate the effect of heart rate changes on dipyridamole echocardiographic tests in patients with coronary artery disease treated with propranolol. We prospectively studied 12 patients (8 men and 4 women; mean age 56.5 +/- 8.7 years) selected by: (a) angiographic evidence of significant coronary artery disease; (b) adequate echocardiographic window; (c) positive dipyridamole echocardiography test results in baseline conditions (step I); (d) test reproducibility in the absence of treatment; (e) negative dipyridamole echocardiography test results after 7 days of treatment with propranolol (120 mg.day-1) in twice divided doses daily (step II). In all patients treated with propranolol, dipyridamole echocardiographic testing was repeated 24 h after the last negative test. In these patients, transoesophageal atrial pacing was performed at peak dipyridamole infusion to increase heart rate to values similar to those observed at baseline (step III). At baseline, heart rate and rate-pressure product were significantly lower in patients treated with propranolol (-20.3% and -22.5% in group II, P < 0.001 vs step I; -24.3% and 26.4% in group III, P < 0.05 vs step I), but the different treatments did not produce significant differences in systolic and diastolic blood pressure. At peak dipyridamole infusion, heart rate and rate-pressure product increased with either placebo or propranolol treatments with respect to baseline, while remaining significantly lower with propranolol as compared to placebo (-29.6% and -29.5% in step II, P < 0.001).(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7498205 TI - Impact of immediate and delayed myocardial scintigraphy on therapeutic decisions in suspected acute myocardial infarction. AB - Early myocardial scintigraphic imaging has become technically feasible in patients admitted to hospital with suspected acute myocardial infarction. After prompt injection of 99mTc-sestamibi, subsequent scintigraphic imaging of perfused myocardium can be performed. During a 5-month period, 237 patients were admitted to the coronary care unit of a district hospital on suspicion of acute ischaemic syndrome, and injection of 99mTc-sestamibi for the performance of myocardial scintigraphy was carried out in 134 patients, on average 2 h after onset of symptoms. The investigation was repeated in 126 patients, on average 18 h after the injection. Three planar views were taken in the coronary care unit with a mobile gamma camera. The prevalence of acute myocardial infarction was 53%. The predictive value at the first scintigraphic imaging for a positive or negative test for myocardial infarction 54% and 56%, respectively. Even exclusion of patients with a previous infarction did not increase the diagnostic validity. The predictive value of a negative test, 77%, at the second scintigraphy was still insufficient to make immediate therapeutic decisions. Myocardial scintigraphy performed early, on suspicion of acute myocardial infarction, cannot therefore be used routinely as a diagnostic test prior to intervention in unselected patients because some 90% of this patient group have myocardial perfusion defects. PMID- 7498206 TI - Acute effect of captopril administration on baroreflex sensitivity in patients with acute myocardial infarction. AB - Depressed baroreflex sensitivity (BRS) after acute myocardial infarction (AMI) is considered an indication of decreased vagal and/or increased sympathetic tone. To determine the effect of angiotensin converting enzyme inhibitors (ACEI) on BRS after AMI we studied 27 patients with a first Q wave AMI, no signs of heart failure and no history of arterial hypertension or diabetes mellitus. An additional group of 10 patients with the same clinical characteristics served as controls. On the 5th day after the onset of AMI, three consecutive boluses of phenylephrine were given intravenously and baseline BRS was taken as the mean slope of the linear regression lines of RR intervals over systolic blood pressure. QT interval was also measured and corrected according to Bazett's formula (QTc). Consequently, a single oral dose of captopril 50 mg or placebo was given to treatment or control group patients, respectively; BRS and QTc were reassessed 1 h later. One hour after captopril administration BRS increased from 5.95 +/- 2.80 to 9.14 +/- 3.46 ms.mmHg-1 (P < 0.0001); QTc increased from 414 +/- 46 to 425 +/- 46 ms (P < 0.0001), systolic blood pressure decreased from 125 +/- 19 to 115 +/- 15 mmHg (P = 0.0002), while heart rate did not change significantly. Baseline BRS was correlated only with age (r = -0.74, P < 0.0001). In the control group, 1 h after placebo, no difference was observed in any variable compared to baseline. Captopril appears to improve BRS immediately in the early phase of AMI. PMID- 7498207 TI - Coronary interventions in Europe 1992. The Working Group on Coronary Circulation of the European Society of Cardiology. AB - The general practice of coronary interventions is influenced by various aspects, traditional, cultural, socioeconomic and personal. The aim of this survey was to collect the data on coronary intervention in all member countries of the European Society of Cardiology. The data from 12 of the 35 national members were missing or grossly incomplete and were therefore excluded from the analysis. CORONARY ANGIOGRAPHY: The total number of coronary angiograms was reported as 683,888, an incidence of 1009 +/- 1021 per million inhabitants (range 9 (Romania) to 3076 (Germany)). Germany (246,115 cases), France (144,754), the United Kingdom (76,296), Italy (45,517) and Spain (43,495) registered 81% of all the coronary angiograms performed. PERCUTANEOUS TRANSLUMINAL CORONARY ANGIOPLASTY: (PTCA) The total number of reported PTCAs was 147,729, which on average accounted for 19 +/- 11% (range 2 (Lithuania) to 53% (Netherlands)) of the coronary angiograms. Most of the PTCAs (82%) were confined to a single vessel. The highest incidence of multivessel PTCA was reported from Slovakia (28%). PTCA took place immediately after the diagnostic study in only 18% of cases. Adjusted per capita, Germany ranked first with 703 PTCAs per million inhabitants, followed by Iceland (619), France (614), Belgium (568) and Austria (485). A major in-hospital complication was reported in 2.5% of the patients undergoing PTCA: 0.4% hospital deaths, 1.0% emergency CABGs and 1.1% myocardial infarctions. NEW DEVICES: Stents were implanted in 3211 patients (2.7% of all PTCA patients), equally distributed between emergency situations (53%) and elective procedures. Other interventional devices were applied in 4133 cases (2.8% of all PTCA cases): directional atherectomy, rotablator, transluminal extraction catheter, laser and Rotacs accounted for 1452, 1232, 55, 558 and 222, respectively. Coronary ultrasound (1350 cases) and coronary angioscopy (373 cases) were rarely performed. CORONARY ARTERY BYPASS GRAFTING (CABG): A total of 63,477 patients underwent CABG in the reporting centres resulting in a PTCA/CABG ratio of 2.3. A significant under reporting of surgery in the participating centres must be assumed. CONCLUSIONS: Although partial reporting might bias conclusions, several findings of this survey are noteworthy: (1) PTCA was a well accepted treatment for coronary artery disease, (2) PTCA was applied more frequently than CABG, (3) there was an extremely wide range of coronary angiography and PTCA performed per million inhabitants, (4) the most common additional procedure was stent implantation, but other new devices were only rarely applied. PMID- 7498208 TI - Angioscopic evaluation of prolonged vs standard balloon inflations during coronary angioplasty. A randomized study. AB - To study the immediate effects of prolonged total balloon inflation during PTCA, 41 patients (44 lesions) with chronic stable angina were randomized for prolonged sequential inflations (three to five inflations of 3 to 5 min each, for a total duration of > or = 12 min, group 1, n = 20 lesions) or 'standard' sequential inflations (three to five inflations of < or = 1 min each, for a total duration of < or = 3 min, group 2, n = 24 lesions). The mean duration of total balloon inflation time was 958 +/- 129 s in group 1 vs 205 +/- 46 s in group 2. Results of angioplasty were assessed on both angiography and percutaneous transluminal coronary angioscopy performed immediately after the procedure. High quality imaging of the coronary lumen and lesion morphology was possible on angioscopy in all patients without any complications. Post-PTCA angiographic percent diameter stenosis was significantly less in group 1 compared to group 2: 26 +/- 10% vs 36 +/- 8% (P < 0.05). On angioscopy, flaps were seen in 16 patients in group 2, but in only six in group 1 (P < 0.02). There was no difference in the incidence of thrombi on angioscopy between the two groups (group 1: nine cases, group 2: 10 cases). Sensitivity of angiographic detection of flaps and thrombi was poor: 10% and 12% respectively. One patient in each group developed a longitudinal dissection, detected on both angiography and angioscopy. CONCLUSIONS: (1) prolonged sequential balloon inflations lead to less residual luminal stenosis after PTCA, with a decreased incidence of intimal flaps in comparison with standard inflations. (2) Post-PTCA transluminal coronary angioscopy is safe and offers better assessment of luminal effects of PTCA than angiography. PMID- 7498209 TI - The effects of ibopamine on glomerular filtration rate and plasma norepinephrine remain preserved during prolonged treatment in patients with congestive heart failure. AB - In acute studies ibopamine, an a selective dopamine agonist, induces moderate increases of GFR and ERPF, and a fall in plasma norepinephrine levels in patients with congestive heart failure (CHF). We evaluated acute and chronic effects of ibopamine on renal haemodynamics, sodium excretion, PRA, plasma aldosterone (ALD) and norepinephrine levels in an open controlled study in 10 patients aged (51-79 years) with mild CHF, NYHA class II-III. All patients used digoxin and frusemide. After a control study day, the second study day involved the administration of 100 mg ibopamine. Subsequently the patients continued to take ibopamine 100 mg three times daily for one month, at which time the chronic effects were measured on the third study day. RESULTS: On the second day ERPF rose from a baseline of 288 +/- 32 to a mean of 308 +/- 32 ml.min-1 x 1.73 m-2 (P < 0.05) during the 4 h after the first administration of ibopamine and GFR rose from 77 +/- 8 to a mean of 84 +/- 8 ml.min-1 x 1.73 m-2 (P < 0.05). The ratio GFR/ERPF, representing the filtration fraction (FF) remained unchanged. On the third study day GFR and ERPF at baseline were similar to those before ibopamine treatment. After the acute on chronic administration we observed an increase in GFR (from 76 +/- 6 to a mean of 85 +/- 7 ml.min-1 x 1.73 m-2 (P < 0.05)), and in ERPF (from 279 +/- 27 to a mean of 293 +/- 29 ml.min-1 x 1.73 m-2 (P < 0.05)). Plasma norepinephrine levels fell from 2.63 +/- 0.48 to 1.92 +/- 0.27 nmol.l-1 (P < 0.05) after the acute administration of ibopamine, and remained unchanged after the acute on chronic ibopamine administration (C: 1.80 +/- 0.42 nmol.l-1). No changes in sodium excretion were observed, either in blood pressure, heart rate, PRA or ALD. We conclude that renal function is preserved during chronic ibopamine treatment and the acute moderate increase of ERPF and GFR after a single dose of 100 mg of ibopamine is still present after one month of treatment with ibopamine in patients with CHF. Ibopamine lowered plasma norepinephrine levels in our patients with CHF, and these values remained unchanged after the acute on chronic administration of ibopamine. PMID- 7498210 TI - Diastolic mitral annular motion in normal subjects and patients with coronary artery disease. AB - The purpose of this study was to evaluate the characteristics of mitral annular motion during diastole in 28 normal subjects, 40 patients with prior myocardial infarction (MI), and 23 patients with coronary artery disease but without prior MI. Mitral annular motion during diastole was obtained from the apex by M-mode echocardiography at the posterior wall of the left ventricle. Determinants of mitral annular excursion during early (MAE-E) and late diastole (MAE-L) were investigated in all subjects. Differences in the MAE-E, MAE-L, and the MAE-L:MAE E ratio were compared among the three patient groups. The Doppler-derived transmitral flow velocity-time integral during early (EI) and late (AI) diastole and mitral annular excursions during diastole were obtained in 55 other patients with a prior MI and in 29 healthy volunteers. The relationships between the MAE L:MAE-E ratio and AI:EI ratio in these two groups were studied. The MAE-E was determined mainly by heart rate and left ventricular ejection fraction (LVEF). The MAE-L was determined only by age. The magnitude of MAE-E was significantly less in patients with a prior MI than in normal subjects (P < 0.01). However, the MAE-L did not differ among the three groups. The MAE-L:MAE-E was higher in patients with a prior MI than in normal subjects (P < 0.05), and was significantly correlated with AI:EI in healthy volunteers (r = 0.65, P < 0.001) and in patients with a prior MI (r = 0.50, P < 0.001).(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7498211 TI - The role of cardiac output response in blood flow distribution during exercise in patients with chronic heart failure. AB - The relationship between blood flow in the working leg and cardiac output (Q) response was determined for patients with chronic heart failure. The ratio of leg blood flow (LBF) to Q (LBF/Q) at the same workload during submaximal exercise was significantly different in three groups of patients classified by peak cardiac index: it was 0.22 +/- 0.08 in the 'satisfactory' Q group vs 0.33 +/- 0.09 in the moderate Q group, and 0.38 +/- 0.08 in the poor Q group (P < 0.01) in the first stage of exercise. delta LBF/delta Q, the slope of change in leg blood flow to Q from rest to peak exercise, was significantly larger in the poor Q group than in the satisfactory Q group (P < 0.01). The intergroup arteriovenous oxygen difference (AVO2D) in organs other than the leg was large, while that in the working leg was small. Leg vascular resistance at submaximal workload was similar in the three groups, but vascular resistance in other organs was significantly higher in the poor Q group than in the satisfactory Q group (P < 0.001). These findings suggested that metabolic vasodilating stimuli in the working leg attenuated excessive vasoconstriction and caused a shift in blood flow from other organs to the working leg under conditions of severely reduced Q response. PMID- 7498212 TI - Application of ultrasound energy for intracardiac ablation of arrhythmias. AB - Ultrasound is a potential energy source for cardiac ablation. Small ultrasound applicators were tested for their ability to create lesions in cardiac tissue. Ultrasound applicators were designed, constructed and tested in canine cardiac tissue in degassed normal saline, and both in vitro and in vivo, lesions were produced by using transducers with frequencies of about 10 MHz. Lesion depth increased with longer duration of energy delivery from 15-60 s, and there was a linear relationship between increasing power and depth of lesions. Seven in vivo experiments in open-chest dogs were performed, and the ultrasound transducers were mounted on the tip of 7-French angiographic catheters. On the epicardium the maximum lesion depth was 9 mm. When the transducer was inserted into the left ventricle, lesions of 8.7 +/- 2.9 mm (n = 4) were produced. It is concluded that an ultrasound transducer mounted on a cardiac catheter can produce lesions that may be useful for ablation of cardiac arrhythmias. PMID- 7498213 TI - Successful radiofrequency catheter ablation of right sided accessory pathways during sustained atrial fibrillation. AB - Up to now there have been no descriptions in the literature of endocardial catheter mapping and successful radiofrequency catheter ablation of the right sided accessory pathway during sustained atrial fibrillation. We attempted ablation of a right posterolateral and a right lateral accessory pathway during sustained atrial fibrillation with rapid anterograde conduction over the accessory pathway in two patients with Wolff-Parkinson-White syndrome. In both patients (aged 47 and 52 years), sustained atrial fibrillation occurred during the electrophysiological study to assess accessory pathway conduction properties. The mean ventricular rate during atrial fibrillation was 180 and 170 beats.min-1, respectively. Both patients were successfully ablated with a single radiofrequency impulse. At the successful ablation sites, a potential of the accessory pathway was consistently recorded preceding the onset of preexcitation in the surface electrogram, by 25 ms in patient 1 and 30 ms in patient 2. The unipolar electrogram recorded from the ablation catheter showed a QS morphology and the accessory pathway potential preceded the intrinsic deflection by 10 ms in both patients. Onset of the ventricular electrogram (patient 1: -20 ms; patient 2: -15 ms) and its activation time (patient 1: -15 ms; patient 2: +5 ms) in relation to the onset of preexcitation in the surface electrocardiogram also indicated the close proximity of the ablation catheter to the accessory pathway. Preexcitation disappeared within 2 s after energy application in both patients. Thus, radiofrequency catheter ablation of right sided accessory pathways during sustained atrial fibrillation is feasible and may obviate the need for medical or electrical cardioversion.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7498214 TI - Analysis of randomness of atrial and ventricular rhythm in atrial fibrillation. AB - The aim of the present study was to examine the relationship between randomness of atrial and ventricular rhythm during atrial fibrillation. Atrial fibrillation was induced in 10 open-chest pigs by application of metacholine on the surface of the right atrium followed by incremental pacing. Local atrial rhythm (AA intervals) was recorded with a bipolar epicardial electrode, and episodes of atrial fibrillation corresponding to 500 ventricular (RR) intervals were selected for analysis. Randomness of the distribution of AA and RR intervals was assessed by autocorrelation. Pearson's test was used for statistical analysis. Random AA and RR interval distribution was observed in nine pigs (P > or = 0.05). In the remaining pig, atrial fibrillation had changed to atrial tachycardia. This was associated with immediate transition of a random to a non-random ventricular rhythm. These findings provide strong circumstantial evidence in support of the contention that randomness of ventricular rhythm during atrial fibrillation is due to randomness of atrial rhythm. PMID- 7498215 TI - Alternations in atrial natriuretic peptide release after DC cardioversion of non valvular chronic atrial fibrillation. AB - The response of atrial natriuretic peptide (ANP) release to haemodynamic influences after cardioversion of atrial fibrillation has not been fully examined. We measured plasma concentrations of ANP and assessed haemodynamic changes 60-120 min after DC cardioversion in 22 patients with non-valvular chronic atrial fibrillation. Passive leg elevation to enhance volume expansion was performed 60 min after DC cardioversion. Sinus rhythm was restored in 18 of the 22 patients (successful DC cardioversion group). The control group consisted of seven patients with non-valvular chronic atrial fibrillation who did not undergo DC cardioversion (atrial fibrillation control group). In the successful DC cardioversion group, the mean pulmonary artery wedge pressure decreased significantly 15 min after cardioversion (P < 0.05) and then remained unchanged. Plasma concentrations of ANP also decreased significantly 15 min after cardioversion (P < 0.05). Furthermore, there was an additional significant decrease in ANP levels for up to 60 min after cardioversion (P < 0.05 from 15 min). Passive leg elevation for 15 min led to an increase in the mean pulmonary artery wedge pressure (P < 0.01) and right atrial pressure (P < 0.05), but did not result in increased plasma concentrations of ANP (47.1 +/- 27.6 vs 43.9 +/- 34.4 pg.ml-1, mean +/- SD, P = ns). In the atrial fibrillation control group, passive leg elevation increased the mean pulmonary artery wedge pressure (P < 0.01), the mean right atrial pressure (P < 0.05) and plasma concentrations of ANP (139.9 +/- 85.8 vs 168.1 +/- 108.2, P < 0.05).(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7498216 TI - Randomized comparison of aspirin and phenprocoumon for prevention of right-sided thromboembolic complications associated with transvenous defibrillation leads. AB - Death due to pulmonary embolism caused by thrombosis formation along a pacing wire has been reported in patients with antibradycardia pacemakers. The purpose of this prospective and randomized study was to evaluate the effects of prophylactically assigned aspirin (100 mg per day) or phenprocoumon (a coumarin derivative) on the incidence of right-sided thromboembolic complications associated with transvenous defibrillation leads. Multiplane transoesophageal echocardiography and pulmonary scintigraphy were performed in 62 consecutive patients immediately and 6 +/- 2 months after transvenous defibrillator implantation. Aspirin or phenprocoumon was administered to the patients in a randomized and parallel manner. By means of multiplane transoesophageal echocardiography and pulmonary scintigraphy no thromboembolic events were observed immediately after transvenous defibrillator implantation. After 6 +/- 2 months, however, multiplane transoesophageal echocardiography disclosed thrombi on the transvenous leads in 13 of 62 patients (21%) despite antithrombotic therapy. At the same time, pulmonary scintigraphy did not reveal any defects compatible with pulmonary embolism. Although thrombi occurred in only four patients treated with phenprocoumon, but in nine patients receiving aspirin, this difference was statistically insignificant. Other variables associated with the occurrence of thrombosis formation were not identified. One fatal intracranial haemorrhage occurred in the phenprocoumon group.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7498217 TI - The chronotropic response of the sinus node to exercise: a new method of analysis and a study of pacemaker patients. AB - In this study, the intercepts and slopes of the regression lines of sinus heart rate response to exercise were examined as a possible means of assessing normal and abnormal chronotropic responses. The regressions were plotted for 223 normal subjects and 93 pacemaker patients during progressive bicycle exercise. The 93 pacemaker patients consisted of two groups, (a) 46 with symptomatic sick sinus syndrome (daytime sinus or nodal bradycardia of < 45.min-1 or atrial asystole > 1.5 s together with other features of the syndrome) and (b) 47 with second- or third-degree AV block (in whom a one-to-one AV relationship was either restored by dual chamber pacing or in whom normal AV conduction was present at the time of exercise). Examination of the regression lines showed them to be sufficiently linear in both normal subjects and pacemaker patients to allow calculation of the intercept and slope values. Multiple regression analysis of these values showed that the AV block group did not differ significantly from the normal individuals. However, in the patients with sick sinus syndrome, the intercept but not the slope values were significantly lower than in the normal subjects (P < 0.01). The patients with sick sinus syndrome appeared to have an abnormality of sinus node function, where the basic rate setting is too low at all levels of exercise, although the acceleration with exercise is normal. Among the sick sinus syndrome patients, only four (9%) had intercept values lying more than 2 standard deviations below that of the normal control group and were deemed to be chronotropically incompetent. Finally, the problems with the various methods used to investigate chronotropic incompetence are reviewed.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7498219 TI - Imaging amyloidosis with radiolabelled SAP. PMID- 7498218 TI - Multiplane transoesophageal echocardiography and morphology of regurgitant mitral valves in surgical repair. AB - BACKGROUND: An essential step in the surgical management of patients with mitral regurgitation, is a thorough understanding of the pathophysiological mechanism. This information can be obtained by multiplane transoesophageal echocardiography which displays all the components of the incompetent valve. METHODS AND RESULTS: Forty-nine patients were scanned intra-operatively by multiplane transoesophageal echocardiography, and findings compared with those at visual inspection during surgery. The pre-operative diagnosis was prolapse of the anterior mitral leaflet in nine patients (sensitivity 100%, specificity 95%), prolapse of the posterior leaflet in 17 patients (sensitivity 100%, specificity 94%) and prolapse of both leaflets in eight patients (sensitivity 87%, specificity 100%). In 11 patients annular dilatation with no abnormalities in mitral leaflet closure or motion was diagnosed (sensitivity 73%, specificity 100%). Two patients had a false-positive diagnosis of prolapse of the anterior leaflet, two others on the posterior leaflet. A prolapse of both leaflets was overlooked in one patient. Multiplane transoesophageal echocardiography scanned the mitral valve, disclosing the extent of pathology along the closure line of leaflets in 88% of patients with mitral valve prolapse. The antero-posterior diameter of the mitral annulus was measured: a diameter over 35 mm indicated annular dilatation. Using this criterion, sensitivity was 89% and specificity 100%. CONCLUSIONS: Multiplane transoesophageal echocardiography enabled components of the mitral valve to be examined systematically, and provided important information on the pathophysiological mechanism of mitral regurgitation before surgical repair. The method also allowed the surgical outcome to be assessed, offering the possibility of optimal repair. PMID- 7498220 TI - Technetium-99m sestamibi kinetics in reperfused canine myocardium. AB - The purpose of the current study was to clarify the myocardial kinetics of technetium-99m sestamibi when the latter is administered during reperfusion. Sestamibi has in the past been given to patients following thrombolytic therapy to document reperfusion and assess salvage. However, the factors which affect sestamibi kinetics during reperfusion are not clearly defined. In this study the left circumflex coronary artery was occluded for 2 h in six dogs (group 1) and for 3 h in six dogs (group 2), followed by reperfusion. Five additional dogs were not reperfused (group 3). Sestamibi was administered during reperfusion in groups 1 and 2, and during ongoing occlusion in group 3. Regional myocardial sestamibi activity was monitored for 3 h using miniature implanted radiation detectors and gamma camera imaging. Group 1 dogs had no infarcts, group 2 had moderate infarcts (mean: 13.9%), and group 3 had large infarcts (mean: 25.2%). Three-hour fractional myocardial clearances were significantly greater for reperfused infarcted (group 2) (0.23 +/- 0.02 SEM) and for nonreperfused infarcted myocardium (group 3) (0.24 +/- 0.02) compared to control (0.10 +/- 0.01) and reperfused noninfarcted myocardium (group 1) (0.07 +/- 0.02; P < 0.01). Quantitative image analysis demonstrated a significant reduction in the left circumflex/left anterior descending count ratios from initial to final scans for group 2 (0.74 +/- 0.03 to 0.65 +/- 0.03, P < 0.05), and a trend towards a reduction in the count ratios from initial to final scans for group 3 (0.38 +/- 0.04 to 0.30 +/- 0.04; P = 0.06).(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7498221 TI - Evaluation in vitro and in rats of 161Tb-DTPA-octreotide, a somatostatin analogue with potential for intraoperative scanning and radiotherapy. AB - The characteristics of terbium-161 diethylene triamine penta-acetic acid (DTPA) labelled octreotide with respect to specific binding to somatostatin (octreotide) receptors on rat brain cortex membranes, biological activity, uptake and excretion by isolated perfused rat livers and metabolism in vivo in normal and tumour-bearing rats were determined and compared to those of indium-111 DTPA octreotide. The results of the binding studies demonstrate that 161Tb-DTPA octreotide is a high-affinity radioligand for somatostatin receptors, with an affinity comparable to that of 111In-DTPA-octreotide. Rat growth hormone secretion inhibition experiments showed that 161Tb-DTPA-octreotide has a similar potency to 111In-DTPA-octreotide. 161Tb-DTPA-octreotide appeared to be taken up even less by the isolated perfused rat liver than 111In-DTPA-octreotide, as almost no tracer disappeared from the perfusion medium. Furthermore, hardly any radioactivity was found in the liver, and excretion into the bile was negligible. The biodistribution studies showed that for octreotide receptor-positive organs, such as pancreas and adrenals, uptake of 161Tb-DTPA-octreotide is lower then that of 111In-DTPA-octreotide. However, as the clearance from the blood of the former compound is faster than that of the latter, the tissue/blood ratio is higher in the case of 161Tb-DTPA-octreotide than with 111In-DTPA-octreotide. Furthermore, these studies demonstrated that the uptake of 161Tb-DTPA-octreotide by the renal tubular cells after glomerular filtration can be reduced by administration of lysine or sodium maleate. Increase in urine production before and during the experiment had no effect on the kidney uptake of 161Tb-DTPA-octreotide.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7498222 TI - Technetium-99m-L,L-ethylenedicysteine scintigraphy in patients with renal disorders. AB - Technetium-99m-L,L-ethylenedicysteine (99mTc-L,L-EC), a new renal imaging agent, was introduced as an alternative to 99mTc-mercaptoacetyltriglycine (MAG3). This radiopharmaceutical can be easily labelled at room temperature and has high radiochemical purity and long stability. The aim of this study was to gain clinical experience in using 99mTc-L,L-EC in normal volunteers and patients. The clearance of this radiopharmaceutical was compared with that of iodine-131 ortho iodohippurate (OIH) in five healthy volunteers. In addition, conventional renogram and whole-body distribution of 99mTc-L,L-EC (40 min and 3 h post injection) were evaluated in these subjects. Subsequently, ten patients with suspected obstructive nephropathy, four with renovascular disorders and two in acute renal failure were imaged. In five patients with impaired renal function both 99mTc-MAG3 and 99mTc-L,L-EC studies were performed. In each case the scintigraphic images and time/activity curves were evaluated and various semiquantitative parameters calculated and compared. No adverse effects were noted during and after 99mTc-L,L-EC scintigraphy. The mean clearance values for 99mTc-L,L-EC and 131I-OIH in volunteers were 504 and 663 ml/min respectively. The total plasma clearance of 99mTc-L,L-EC was about 75.8% of the 131I-OIH value. In volunteers the parenchymal transit time index, whole kidney transit time index and mean parenchymal transit time for 99mTc-L,L-EC were 63 s, 124 s and 175 s respectively. The mean time to peak activity was 235 s and the time from peak to 50% of peak activity was 402 s.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7498224 TI - A quantitative approach to technetium-99m ethyl cysteinate dimer: a comparison with technetium-99m hexamethylpropylene amine oxime. AB - To develop non-invasive regional cerebral blood flow (rCBF) measurements using technetium-99m ethyl cysteinate dimer (99mTc-ECD) and single-photon emission tomography (SPET), the same graphical analysis as was described in our previous reports using technetium-99m hexamethylpropylene amine oxime (99mTc-HMPAO) was applied to time-activity data for the aortic arch and brain hemispheres after intravenous injection of 99mTc-ECD. Hemispherical brain perfusion indices (BPI) for 99mTc-ECD showed a highly significant correlation (n = 22, r = 0.935, P = 0.0001) with those for 99mTc-HMPAO in 11 patients who underwent both tracer studies. Using both linear regression line equations between 99mTc-ECD BPI and 99mTc-HMPAO BPI and between 99mTc-HMPAO BPI and mean cerebral blood flow (CBF) values obtained from a xenon-133 inhalation SPET method in a previous study, 99mTc-ECD BPI was converted to 133Xe CBF values (y = 2.60 chi + 9.8). Then raw SPET images of 99mTc-ECD were converted to rCBF maps using Lassen's correction algorithm. In this algorithm, the correction factor alpha was fixed to 1.5, 2.6 and infinite. In the comparison of rCBF values for 99mTc-ECD SPET with those for 99mTc-HMPAO SPET in 396 regions of interest in the aforementioned 11 patients, the fixed correction factor alpha of 2.6 gave nearly the same rCBF values for 99mTc-ECD (50.1 +/- 16.9 ml/100 g/min, mean +/- SD) as for 99mTc-HMPAO (49.9 +/- 17.3 ml/100 g/min). In conclusion, the same non-invasive method as has been used in 99mTc-HMPAO studies is applicable to a 99mTc-ECD study for the measurement of rCBF without any blood sampling. PMID- 7498223 TI - Is planar thallium-201/fluorine-18 fluorodeoxyglucose imaging a reasonable clinical alternative to positron emission tomographic myocardial viability scanning? AB - This comparative study was performed to determine whether a conventional planar gamma camera optimised for 511-keV imaging can reliably assess myocardial viability using the fluorine-18 fluorodeoxyglucose (FDG) metabolic tracer previously developed for positron emission tomography (PET). Twenty-seven patients with severe ischaemic cardiomyopathy (mean left ventricular ejection fraction: 20% +/- 9%) having clinically indicated nitrogen-13 ammonia/FDG PET myocardial viability studies consented to resting, four-view, planar myocardial thallium-201 perfusion and FDG metabolism imaging. The resultant PET and planar perfusion/metabolism images (PPI) were independently assessed for FDG defect size and perfusion/metabolism mismatch, using a four-point scale, in each of four vascular regions: apex, circumflex, left anterior and posterior descending coronary artery territories. Of 108 regions, 106 were evaluable (two not assessed by PET). There was complete agreement in 70% of coronary vascular territories, giving an unweighted kappa score of 0.56. Moreover, in 94% of segments agreement was within one grade. Interestingly, six of the seven differences of more than one grade occurred in the circumflex coronary territory, which was also the only region for which planar positron imaging underestimated FDG defect size. Three of four moderate areas of perfusion/metabolism mismatch seen with PET were also seen on PPI. PPI showed three small regions of mismatch not seen on PET, whilst the reverse occurred with one other small region of mismatch. Thus, for this PET protocol, PPI provides very similar information on the extent of regional FDG uptake and occurrence of mismatch.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7498225 TI - Contribution of phagocytic cells and bacteria to the accumulation of technetium 99m labelled polyclonal human immunoglobulin at sites of inflammation. AB - The purpose of this study was to assess the contribution of phagocytic cells and bacteria to the accumulation of technetium-99m labelled polyclonal human immunoglobulin (HIG) at sites of inflammation. Mice were intraperitoneally injected with Staphylococcus aureus (SA animals), with heat-inactivated newborn calf serum (NBCS, to mimic a non-bacterial inflammation) or with physiological saline (controls); 1 h thereafter they received HIG. At various intervals after the administration of HIG the mice were killed, and the percentages of radioactivity in the peritoneal effluent and attached to the cellular and bacterial fraction thereof were established. Furthermore, the total number of cells and that of bacteria in the fluid were quantitated. The percentage of activity in the effluent in the SA animals was (P < 0.02) higher than those in the NBCS-injected animals and controls from 4 h onwards. In all groups of mice this percentage was highest at 4 h and decreased (P < 0.01) afterwards. The percentage of cell-bound activity and the total number of cells remained fairly constant or increased with time in the SA animals (P < 0.01). The bacteria-bound activity remained rather constant throughout the experiment and ranged between 4% and 6%. In the SA-infected animals the percentage of cell-bound activity was correlated with the total number of cells (macrophages but especially neutrophils) but even more strongly with the number of cell-associated bacteria. In the NBCS-injected animals a correlation was demonstrated between the cell bound activity and the total number of cells (only neutrophils).(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7498226 TI - Radioimmunoscintigraphy of ovarian tumours with technetium-99m labelled monoclonal antibody-170: first clinical experiences. AB - The recently developed technetium-99m-labelled monoclonal antibody-170 (MAb-170) was designed for diagnostic use in patients suffering from gynaecological adenocarcinoma. Following in vitro studies which showed immunoreactivity of this antibody to more than 90% of human adenocarcinomas, the present investigation was initiated to verify its usefulness for radioimmunoscintigraphy of ovarian tumours. Most of the 30 patients participating in this study underwent immunoscintigraphy prior to first-look surgery. Biokinetic evaluation in two patients showed a plasma half-time of 18.9 h (mean value, n = 2, r = 0.98) and a biexponential total body curve with values of 7.7 h and 17 days (r = 0.98). The mean 24-h urinary excretion was 12% of the injected dose. Radioimmunoscintigraphy using the MAb-170 recognised 12 of 13 cases of adenocarcinoma of the ovaries, corresponding to an overall sensitivity of 92.3%. Specificity was 94.1% (16/17). The calculation of accuracy yielded a figure of 93.3% (28/30). Of 33 known lesions, 26 were visualised successfully; thus the locoregional sensitivity was 78.8%. Of 29 benign tumour sites, 28 showed no evidence of tracer accumulation, corresponding to a locoregional specificity of 96.6%. The smallest lesion visualised was an adenocarcinoma of the corpus uteri with a diameter of 1.5 cm. Technetium-99m labelled MAb-170 is a promising new radiopharmaceutical for immunoscintigraphy of ovarian adenocarcinoma. PMID- 7498227 TI - Effect of tissue heterogeneity on quantification in positron emission tomography. AB - As a result of the limited spatial resolution of positron emission tomographic scanners, the measurements of physiological parameters are compromised by tissue heterogeneity. The effect of tissue heterogeneity on a number of parameters was studied by simulation and an analytical method. Five common tracer models were assessed. The input and tissue response functions were assumed to be free from noise and systematic errors. The kinetic model was assumed to be perfect. Two components with different kinetics were mixed in different proportions and contrast with respect to the model parameters. Different experimental protocols were investigated. Of three methods investigated for the measurement of cerebral blood flow (CBF) (steady state, dynamic, integral), the second one was least sensitive to errors caused by tissue heterogeneity and the main effect was an underestimation of the distribution volume. With the steady state method, errors in oxygen extraction fraction caused by tissue heterogeneity were always found to be less than the corresponding errors in CBF. For myocardial blood flow the steady state method was found to perform better than the bolus method. The net accumulation of substrate (i.e. rCMRglc in the case of glucose analogs) was found to be comparatively insensitive to tissue heterogeneity. Individual rate constants such as k2 and k3 for efflux and metabolism of the substrate in the pool of unmetabolized substrate in the tissue, respectively, were found to be more sensitive. In studies of radioligand binding, using only tracer doses, the effect of tissue heterogeneity on the parameter kon.Bmax could be considerable. In studies of radioligand binding using a protocol with two experiments, one with high and one with low specific activity, Bmax was found to be insensitive while Kd was very sensitive to tissue heterogeneity. PMID- 7498228 TI - Simultaneous administration of 111In-human immunoglobulin and 99mTc-HMPAO labelled leucocytes in inflammatory bowel disease. AB - Technetium-99m hexamethylpropylene amine oxime (HMPAO) labelled leucocytes and indium-111 polyclonal immunoglobulin (IgG) were simultaneously injected into a group of 27 patients routinely referred for the investigation of inflammatory bowel disease (IBD). Ten-minute anterior abdomen and tail on detector views were obtained at 30 min, 4 h and 24 h p.i. of both tracers. The diagnosis of IBD was obtained in all cases by endoscopy with biopsy and/or surgery. Images were blindly evaluated by two experienced observers who only knew of the clinical suspicion of IBD. IBD was confirmed in 20 patients (12 with Crohn's disease and eight with ulcerative colitis). Sensitivity, specificity and accuracy were 100%, 85% and 96% respectively for labelled leucocytes and 70%, 85% and 74% for IgG. Both IgG and leucocyte scans were normal in six out of seven patients in whom a diagnosis of IBD was excluded; the remaining patient, with ischaemic colitis, was falsely positive with both agents. As far as disease extension is concerned, the IgG study localized 27 diseased segments, whereas 49 were seen with the leucocyte study. Eighty-four segments were normal and 25 showed tracer uptake with both agents. Twenty-four were positive only with the leucocyte study and two were positive only with the IgG study. Agreement between the agents was 80.7%. These results confirm that 111In-human polyclonal scintigraphy is less sensitive than 99mTc-HMPAO scintigraphy both for the diagnosis of IBD and in the evaluation of disease extension. Nevertheless, if leucocyte labelling is not available, labelled IgG can be used only for diagnostic purposes. PMID- 7498229 TI - Optimum tomographic reconstruction parameters for HMPAO brain SPET imaging: a practical approach based on subjective and objective indexes. AB - The purpose of this study was to define an optimal strategy for the tomographic reconstruction procedure in routine brain single-photon emission tomography (SPET) studies, including the number of projections, filter function and matrix size. A set of projection data with different count densities was obtained from a technetium-99m hexamethylpropylene amine oxime (99mTc-HMPAO) brain SPET acquisition from one volunteer. The projections were reconstructed with different filters and the quality of the reconstructed images was determined using both a subjective observer rating score and the Gilbert index. For each count density, the observers' choice corresponded to images with the lowest Gilbert index. The noise level in brain SPET sections was estimated and correlated with the fractal dimension. The results of this study indicate that although noise represents a fundamental component of brain SPET imaging, image quality also depends on the reconstructed spatial resolution. Image quality is satisfactorily described by fractal dimension. In addition the optimal filter function depends on the available count density. For high count levels, optimal reconstruction may be obtained by using a high-resolution matrix and a slightly smoother reconstruction filter. When count densities are low, best results are obtained by using a low resolution matrix and a sharper filter. Finally, this study suggests that image quality is not influenced by the number of projections for equivalent count densities. These results were confirmed by 30 HMPAO brain SPET studies acquired in a routine clinical setting. PMID- 7498230 TI - Iodine-131 Hippuran for the estimation of renal plasma flow: requirements for radiochemical purity. AB - For many years iodide-131 Hippuran has been used as a tracer to measure effective renal plasma flow (ERPF). Because of the low renal clearance of free 131I-iodide and the inability to count it separately from 131I-Hippuran, free 131I-iodide will lower the calculated 131I-Hippuran clearance, resulting in a lower estimated ER-PF. This study was performed to establish the maximum allowable radiochemical impurity of free 131I-iodide in 131I-Hippuran preparations for ERPF measurements in continuous clearance studies. A known amount of 123I-iodide was added to the (131I-iodide-free) 131I-Hippuran solution used for continuous infusion clearance studies in nine patients. 123I-iodide activity was used because it can be counted separately from 131I-Hippuran in the infusion solutions and plasma samples while it behaves exactly like 131I-iodide, so that the results obtained with 123I iodide can be extrapolated to 131I-iodide. After performing the clearance studies, the ERPF was calculated firstly with 131I-Hippuran activity only (= true ERPF) and secondly including the free radioactive iodide activity (= false ERPF) in the clearance formula. As expected, if free 131I-iodide is present in the infusion solution, its concentration in plasma will be highest at the end of the clearance study. The 131I-iodide concentration in plasma relative to the 131I Hippuran concentration will be higher in patients with high ERPF values.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7498231 TI - Iodine-123 labelled N-(2-fluoroethyl)-2 beta-carbomethoxy-3 beta-(4 iodophenyl)nortropane for dopamine transporter imaging in the living human brain. AB - There are several cocaine analogs which have potential for imaging the dopamine transporters (DAT). Earlier studies have shown that iodine-123 labelled 2 beta carbomethoxy-3 beta-(4-iodophenyl)tropane ([123I]beta-CIT) and N-(3-fluoropropyl) 2 beta-carbomethoxy-3 beta-(4-iodophenyl)nortropane ([123I]beta-CIT-FP) are promising DAT imaging agents in the living human brain with single-photon emission tomography (SPET). Here we report a pilot comparison of [123I]beta-CIT and [123I]beta-CIT-FP with a new tropane derivative, [123I]N-(2-fluoroethyl)-2 beta-carbomethoxy-3 beta-(4-iodophenyl)nortropane ([123I]beta-CIT-FE), using SPET imaging in four healthy male subjects. Peak uptake of [123I]beta-CIT-FE into the basal ganglia occurred very rapidly (0.5 h after injection of tracer), after which the striatal washout obeyed a bi-exponential form. The specific DAT binding of [123I]beta-CIT-FE into the basal ganglia was somewhat less (0.785 +/- 0.117) than that of [123I]beta-CIT (0.922 +/- 0.004) or [123I]beta-CIT-FP (0.813 +/- 0.047). All these tracers have excellent imaging quality in healthy control subjects. However, the relatively fast washout of [123I]beta-CIT-FE and low temporal resolution of older SPET cameras may limit the use of this tracer to the measurement of the DAT density. PMID- 7498232 TI - Technetium-99m-tetrofosmin uptake in malignant lung tumours. AB - Technetium-99m-tetrofosmin is a new myocardial imaging agent which has yielded promising results compared to thallium-201. The tumour-seeking properties of the routinely used cardiac radiopharmaceuticals 201TI and 99mTc methoxyisobutylisonitrile are well known. Here we report the results of a pilot study demonstrating 99mTc-tetrofosmin uptake in malignant lung tumours. Five patients with bronchial carcinoma, each in different stages of chemo- or radiotherapy, were imaged. Dynamic and static acquisitions were performed to evaluate the uptake and kinetics of 99mTc-tetrofosmin in the lesions. In four of the five patients localized tumour uptake of 99mTc-tetrofosmin was observed. Time to peak tumour activity and tracer washout in the tumour, myocardium and contralateral normal lung at 30 min post injection (p.i.) were determined. Tumour/normal lung, heart/tumour and heart/contralateral normal lung ratios were calculated for 5-10, 25-30 and 85-90 min p.i. The peak concentration in all tumours was reached at the end of the first minute. The mean tumour and contralateral normal lung washout rates of 99mTc-tetrofosmin at 30 min p.i. were 18.3% +/- 9.2% and 19.5% +/- 5.85% respectively. The tumour/contralateral normal lung ratio remained higher than 1.25 until 90 min p.i. in all four patients. It is concluded that 99mTc-tetrofosmin seems to be of value in lung tumour imaging, although larger studies are necessary to ascertain its sensitivity, specificity and usefulness in clinical practice. PMID- 7498233 TI - Experience with the iodine-123 and technetium-99m labelled anti-granulocyte antibody MAb47: a comparison of labelling methods. AB - Four different methods of radiolabelling the anti-granulocyte monoclonal antibody MAb47 were compared and their influence on diagnostic value studied. The best clinical images were obtained following labelling with iodine-123 by the Iodogen method and direct labelling with technetium-99m after tris-(carboxyethyl) phosphine treatment of MAb47 to achieve disulphide bridge reduction. 99mTc labelling using a specific ligand (MAb47-mtp), or a second method involving direct reduction with mercaptoethanol, led to an increased background activity in clinical studies, thus impeding the diagnosis of chronic disease. Fresh infections were clearly localized by all four preparations. The elimination of the activity from the blood was slower in the case of the iodinated MAb47, while the collected urine samples showed an excretion of about 10% of the injected activity per day independent of the labelling method. The results in terms of sensitivity and specificity were rather similar for all labelling methods and ranged from 90% to 99%. PMID- 7498234 TI - Traditional receptor theory and its application to neuroreceptor measurements in functional imaging. AB - The mathematical, biological discipline of receptor pharmacology and the graphical methods of describing receptor behaviour evolved over a century of painstaking observation and model testing. Laws regarding in vitro theories are classically considered inoperative in vivo. Nevertheless, functional imaging techniques have rapidly evolved to allow receptor measurement and rules of thumb have been developed which clearly prove valid receptor parameters can be derived from functional imaging studies. The field is evolving so rapidly now that nuclear medicine researchers are in danger in applying these techniques without recourse to an understanding of the orthodox discipline of receptor pharmacology. This review attempts to document the basis of receptor pharmacology and to give an account of the theoretical and practical basis on which this can be applied in vivo. The review is targeted towards single-photon emission tomography because of the rapid growth in the area, but many parts draw on the literature relating to positron emission tompgraphy since the first translation of in vitro to in vivo measurement was performed with this technique. PMID- 7498235 TI - n-3 polyunsaturated fatty acids: update 1995. AB - Epidemiologic and biochemical studies have suggested an anti-inflammatory effect of n-3 fatty acids. Beneficial therapeutic effects reported from small patient groups need to be confirmed in large-cohort controlled clinical trials. There is a growing number of clinical trials of n-3 fatty acid supplementation in disease. Clinical benefits have been moderate in patients with rheumatoid arthritis and with arterial hypertension. Clearly negative results have been reported during the past 2 years for patients with lupus nephritis and for patients with psoriasis or with atopic dermatitis. Such trials have now been completed. For patients with coronary artery disease following coronary angioplasty, earlier results of a large meta-analysis, could not be confirmed. For patients with IgA nephropathy and for patients following kidney transplantation, a clear benefit was seen in patients receiving fish oil. These promising results are currently pursued in follow-up phase III clinical trials. PMID- 7498236 TI - Detection improvement of cytomegalovirus antigen in human peripheral blood using monoclonal antibodies and automated reading of cell preparations. AB - One of the major drawbacks in cytomegalovirus (CMV)-antigenaemia detection for diagnosis of active CMV infection is the low number of CMV-antigen positive cells present in peripheral blood. It is therefore necessary to screen large numbers of peripheral blood granulocytes to find only a few antigen-positive cells. We have optimized this detection by testing several monoclonal antibodies (mAb) to CMV antigens (mAbs C10/C11, C12, BM222, E13 and SL20). In total 550 blood samples from 40 patients were investigated. More blood samples were found positive with mAb C12 than with the other mAbs. Also the average number of positive cells per slide was highest for mAb C12. Furthermore, duplicate slides were examined automatically using an image analysis system (LEYTAS) and compared to visual detection (cytospin slides). The detection sensitivity of both screening methods was compared for mAb C12. In total 360 slides were analysed, from positive as well as negative blood samples. The sensitivity of the automated screening was 93% and for the visual evaluation of the cytospin slides 73%. In conclusion, mAb C12 was the most suitable of the mAbs tested for detection of antigenaemia, and automatic detection of CMV antigenaemia with image analysis of slides is a sensitive method due to the large numbers of cells that can be screened. PMID- 7498237 TI - Lipoprotein (a) in patients with hyperlipidaemia. AB - Lipoprotein (a) [Lp(a)] is an atherogenic lipoprotein which is similar in structure to, but metabolically distinct from, LDL. Factors modulating plasma Lp(a) concentrations are poorly understood. We hypothesized that patients with hyperlipidaemia have elevated Lp(a) levels and determined the phenotype, concentration and distribution of Lp(a) in a group of hyperlipidaemic patients (n = 107) compared with a control group (n = 128). Lp(a) concentrations were significantly increased in the hyperlipidaemic patients (mean, 34 +/- 4 mg dL-1; median, 19 mg dL-1) as compared with the controls (20 +/- 3 mg dL-1; 9 mg dL-1) (P < 0.01). Interestingly, after dividing the patients into one group with elevated cholesterol (> 200 mg dL-1) (n = 44) and another group with elevated triglycerides (> 200 mg dL-1) (n = 51) we found that Lp(a) concentrations were 2.3-fold higher in the high cholesterol patients (mean, 45 +/- 5; median, 41 mg dL-1) compared to the high triglyceride subjects (20 +/- 4; 8 mg dL-1) (P < 0.01). Furthermore, a negative correlation between triglyceride and Lp(a) plasma concentrations was found in patients exhibiting triglyceride levels > 300 mg dL-1 (r = -0.41, P = 0.04, n = 36) and with triglycerides > 400 mg dL-1 (r = -0.52, P = 0.03, n = 17). These data indicate that plasma Lp(a) concentrations are elevated in hyperlipidaemia if the patients have high cholesterol levels, whereas Lp(a) is normal to low in patients with elevated triglycerides.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7498238 TI - Increased serum interleukin-1 beta during treatment of hyperthyroidism with antithyroid drugs. AB - Serum interleukin-1 beta (IL-1 beta) and soluble interleukin-2 receptor (sIL-2R) levels were examined in patients with hyperthyroidism due to Graves' disease (GD) and toxic nodular goitre (TNG) before and during antithyroid drug therapy. A total of 32 patients were studied; 23 patients (14 with GD and nine with TNG) were in a hyperthyroid state (group A) and nine patients (four with GD and five with TNG) were in a euthyroid state, under carbimazole or methimazole treatment (group B). Ten hyperthyroid patients from group A (seven with GD and three with TNG) were also examined while euthyroid on treatment (Subgroup A). Serum was taken from all patients for the measurement of sIL-2R, IL-1 beta, total T4 (TT4), total T3 (TT3) and TSH concentrations. The results were compared with those from 30 normal controls. Serum sIL-2R levels were higher in Group A (671.3 +/- 74.0 U mL-1, mean +/- SE), than in Group B (214.1 +/- 61.8 U mL-1) and controls (149 +/- 14.8 U mL-1), P < 0.001. Similarly, the subgroup of 10 patients had higher levels of sIL-2R during the hyperthyroid phase than while euthyroid (P < 0.001). There was a positive correlation between sIL-2R values and levels of T4 and T3. In contrast, serum IL-1 beta levels were higher in Group B patients (197.5 +/- 39.2 pg mL-1) compared with those in Group A (66.5 +/- 17 pg mL-1, P < 0.01).(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7498239 TI - The myeloic related protein MRP8/14 (27E10 antigen)--usefulness as a potential marker for disease activity in ulcerative colitis and putative biological function. AB - MRP8, MRP14 and their heterodimer MRP8/14 (27E10 antigen) are myeloic related proteins which have been shown to have a major role in inflammatory and immunological responses. In the present study monospecific antibodies against MRPs were used to investigate immunohistochemically the distribution of these proteins in routinely processed bowel tissues from 23 patients with ulcerative colitis (UC). MRP8, MRP14 and their heterocomplex MRP8/14 were demonstrated in the majority of granulocytes and macrophages in tissues of patients with active UC. Furthermore by employing the ELISA technique we measured MRP8/14 serum levels in 62 patients with UC and the results were compared with those for healthy controls. Disease activities were determined by established clinical activity indices. Serum MRP8/14 concentrations were significantly (P < 0.0001) increased in patients with active ulcerative colitis. No enhancement of serum levels were found for MRP14 and MRP8 alone, respectively. The follow-up of individual patients with initially active disease showed a decrease of MRP8/14 serum levels in parallel with clinical improvement following the start of therapy. It is thus concluded that MRP8/14 accurately reflects the degree of disease activity in UC. Further, possible biological function of MRPs seems to be associated with the heterodimeric form (27E10 antigen) rather than with individual proteins. Our morphological results confirm the finding of enhanced MRP8/14 serum levels in patients with active UC. PMID- 7498240 TI - Glutathione, glutathione utilizing enzymes and thioltransferase in platelets of insulin-dependent diabetic patients: relation with platelet aggregation and with microangiopatic complications. AB - Reduced glutathione (GSH) and activity of GSH related enzymes play a key role in defence against oxygen free radicals, whose production is, as known, raised in patients affected by diabetes mellitus, and at the same time they may contribute to the process of platelet aggregation. The purpose of this study was to evaluate GSH levels and activity of glutathione peroxidase (GSH-Px), glutathione reductase (GSSG-Red), glutathione transferase (GSH-Tr), glucose-6-phosphate-dehydrogenase (G6PDH), and thioltransferase (TT) in platelets of insulin-dependent diabetic patients in fair metabolic control (mean glycated haemoglobin: 6.5%), as related to presence of retinopathy, neuropathy or nephropathy and to platelet aggregation by arachidonic acid (AA) in vitro. Mean effective dose (ED50) of AA was on average significantly lower in the group of insulin-dependent diabetic patients (0.41 +/- 0.02 mM (SEM), n = 46) as compared with that of control subjects strictly matched for age, sex and weight (0.77 +/- 0.02, n = 51; P = 0.0001). Mean platelet GSH as well as the activity of GSH related enzymes expressed as geometric mean (95% confidence intervals) were similar in diabetic patients and in controls, except for GSSG-Red whose activity was significantly higher in diabetic subjects (28.5 (14.4-57.5) mU 10(-9) platelets vs. 20.3 (8.7-56) mU 10( 9) platelets; P = 0.01). In the diabetic group TT was reduced when compared with healthy controls (3.8 (0.9-12.2) mU 10(-9) platelets vs. 6 (1.6-26.1) mU 10(-9) platelets; P = 0.04).(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7498241 TI - Renal and adrenal mechanisms in cyclosporine-induced hyperkalaemia after renal transplantation. AB - CsA associated hyperkalaemia was investigated in 24 renal transplant recipients 6 months after transplantation. 51Cr-EDTA-, PAH-, lithium and sodium clearances, 24 h urinary creatinine and potassium excretions, plasma renin activity and aldosterone concentrations were measured. Transtubular potassium concentration gradient (TTKG) was calculated. An ACTH test was performed to document adrenal function. Eleven patients had hyperkalaemia. The TTKGs were low normal or reduced in both normo- and hyperkalaemic patients implying inhibition of K+ secretion. The hyperkalaemic patients received more CsA (mean dose 21.3 vs. 9.7 mg kg-1d-1, P = 0.01), and had lower lithium clearances (mean 9.9 vs. 17.0 mL min-1 1.73 m-2, P < 0.05). Adrenal function had no clear effect. Serum potassium concentration correlated with CsA dose (r = 0.773, P < 0.001) and inversely with lithium clearance (r = -0.568, P < 0.01) suggesting that CsA induced decrease in distal tubular flow rate reduced K+ excretion. Hyperkalaemia was not fully explained by renal mechanisms. PMID- 7498242 TI - Inhibition of ileal bile acid transport by cyclosporin A in rat. AB - Chronic administration of cyclosporin A may induce cholestasis and this effect has been attributed to impaired hepatic bile salt synthesis, metabolism and transport. We investigated the effect of cyclosporin A on intestinal absorption of bile acids in the ileum of rat. Ileal bile acid absorption was measured by in vivo intestinal perfusion with cyclosporin A and the solvent Cremophor EL. During ileal perfusion with 25 microM glycocholic acid, the concentration of 2.8 mM cyclosporin A inhibited intestinal bile acid absorption on average by 34%. Additional experiments were performed with everted gut sacs of the distal ileum to evaluate active absorption. A dose and time dependent inhibition of the active intestinal absorption of bile acids was found, with a 50% transport inhibition at an average cyclosporin A concentration of 2.69 mmol L-1. Thus, cyclosporin A inhibits the active intestinal absorption of bile acids which may influence bile acid synthesis, turnover and secretion and may contribute to cyclosporin A induced cholestasis. PMID- 7498243 TI - Interferon-gamma and urine neopterin in attacks of the hyperimmunoglobulinaemia D and periodic fever syndrome. AB - The hyperimmunoglobulinaemia D and periodic fever (hyper-IgD) syndrome is typified by recurrent unpredictable febrile attacks with abdominal pain, joint involvement (arthralgias/arthritis), headache, skin lesions and a polyclonal elevation of serum IgD (> 100 U mL-1). Interferon-gamma (IFN-gamma) is a major proinflammatory cytokine which could play a role in the pathogenesis of the attacks. There is a need for parameters (if possible non-invasive) to monitor disease activity. A potential candidate is neopterin which is released by monocytes/macrophages when stimulated with IFN-gamma, excreted unchanged in urine, and appears to be an early and sensitive marker for activation of the immune system. We measured rectal body temperature, serum IFN-gamma, and urine neopterin in 10 hyper-IgD patients both during and between attacks. The body temperature rose to a mean of 38.9 degrees C on the first day of the attack and normalized within 5 days. Serum IFN-gamma during the first day of the attack was 2.98 IU mL-1 and was significantly lower during remissions. The urine neopterin excretion was 268 +/- 170 mumol mol-1 creatinine between attacks and was significantly increased to 638 +/- 275 mumol mol-1 creatinine on the first day of symptoms. Maximal urine neopterin values were reached on the fourth day of the attack (1051 +/- 387 mumol mol-1 creatinine) and excretion gradually declined and attained values below 400 mumol mol-1 creatinine after 9 days. There was a good correlation between serum IFN-gamma and urine neopterin.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7498244 TI - Neutrophil dysfunction and increased susceptibility to infection. AB - A critical evaluation of 3 years' experience using laboratory screening to detect neutrophil dysfunction is described. Neutrophil dysfunctions in patients with recurrent bacterial infections were investigated by using the following screening tests: (1) neutrophil chemotaxis towards N-formylmethionyl peptides (FMLP) and the complement fragment C5a; (2) neutrophil production of superoxide anions (O2-) in response to phorbol myristate acetate and opsonized zymosan particles; and (3) examination of May-Grunwald and myeloperoxidase cytochemical staining of peripheral blood smears. These tests were carried out in 100 patients suffering from infections and suspected of having altered neutrophil functional competence. A minority of patients was found to have well defined neutrophil dysfunction syndromes: chronic granulomatous disease (four cases), Chediak-Higashi disease (one case) and myeloperoxidase deficiency (one case). Of the remaining 94 patients, in whom infections localized to airways and/or skin predominated, 53 cases were found to have impaired chemotaxis (41 cases) or partial defects of the O2- production. Defects of chemotaxis toward FMLP and those towards both FLMP and C5a were the most frequent abnormalities. No defect was found in the other 41 patients. Moreover, impaired neutrophil chemotaxis was found in some patients with selective IgA deficiency (five cases) or immotile cilia syndrome (seven cases). The results suggest that (a) additional screening tests are required to ameliorate the efficiency of the diagnostic work-up of the patients suspected to have neutrophil dysfunction; and (b) further evaluation, also at the molecular level, should be considered at least in selected cases of non-classified neutrophil dysfunction in order to clarify diagnosis and plan rational therapeutic strategies. PMID- 7498245 TI - Determinants of insulin-stimulated skeletal muscle glycogen metabolism in man. AB - To examine factors which influence skeletal muscle glycogen synthesis in man, we related insulin sensitivity measured by euglycaemic insulin clamp in 43 healthy males to muscle glycogen synthase (GS) activity, GS protein content (Western blot), glycogen concentrations and fibre composition. Insulin increased muscle glycogen content (P < 0.05) and the change in glycogen content correlated with the GS protein content (r = 0.90, P = 0.01). GS protein concentration correlated inversely with age (r = -0.69, P = 0.04). Non-oxidative glucose disposal was inversely related to per cent type 2B fibres (r = -0.52, P < 0.05). The influence of age on these relationships was separately studied in young (n = 12, age = 26 +/- 2 years) and elderly (n = 15, age = 56 +/- 2 years) males. Insulin increased GS activity significantly only in young subjects (from 17.8 +/- 3.0 to 25.3 +/- 3.2 nmol mg protein-1 min-1; P = 0.015). GS activity and non-oxidative glucose disposal correlated in young (r = 0.69, P = 0.01) but not in the elderly (r = 0.064, P = 0.82) males, and this relationship was not influenced by the degree of obesity. In conclusion, muscle fibre type and GS activity are both determinants of muscle glycogen metabolism in healthy, normoglycaemic males. The close relationship between non-oxidative glucose metabolism and GS activity in young males is altered in ageing. PMID- 7498246 TI - Cellular processing of apolipoprotein B-containing lipoproteins from young post infarction patients and healthy controls. AB - 3-Hydroxy-3-methylglutaryl-coenzyme A (HMG-CoA) reductase activity was measured in fibroblasts incubated with large (Sf > 60) and small (Sf 20-60) very low density lipoprotein (VLDL), intermediate density lipoprotein (IDL) and low density lipoprotein (LDL) particles, at similar protein concentrations, from young post-infarction patients and healthy controls. The results showed that apolipoprotein (apo) B-containing lipoproteins (VLDL, IDL, LDL) from patients suppressed HMG-CoA reductase activity to a similar extent compared to apo B containing lipoproteins from controls. When all subjects taken together were grouped according to triglyceride levels, it was found that small VLDL from hypertriglyceridaemic individuals suppressed the HMG-CoA reductase activity more than small VLDL from normotriglyceridaemic individuals. The opposite pattern was seen for LDL. The lipoprotein composition was related to the respective HMG-CoA reductase activity. In addition to a positive association between the cholesterol content of small VLDL and LDL, and the inhibition of HMG-CoA reductase activity, the apo C I and C II content of small VLDL and IDL was inversely related to the suppression of HMG-CoA reductase activity. This study shows that the cellular processing of apo B-containing lipoproteins in young post-infarction patients and healthy controls is heterogeneous and dependent on the composition of the lipoprotein. PMID- 7498247 TI - Direct assessment of extracellular water volume by the bromide-dilution method in growth hormone-deficient adults. AB - Body composition and water content were assessed in 36 growth hormone deficient (GHD) patients (M:23, F:13) and 14 controls (M:7, F:7) using anthropometry, hydrodensitometry, bioelectrical impedance measurement and the bromide-dilution method, with which extracellular water volume can be measured directly. GHD patients, compared to controls, did not differ in extracellular water volume, in hydration state and water distribution. A higher BMI (P = 0.002) and a higher body fat mass (P < 0.0001) were found in the patients. Based on circumference measurements, the GHD patients had a higher waist/hip ratio (P = 0.0001). This study shows that GHD patients, in comparison with healthy controls, have a normal extracellular water volume. The finding that extracellular water volume is within the normal range in GHD patients may be clinically relevant as it is well known that in the first months after initiating GH-replacement therapy, most patients have signs and symptoms of excessive water retention. PMID- 7498248 TI - Effects of nifedipine and Bay K 8644 on myotropic responses in aortic rings of pregnant rats. AB - The hypothesis that Ca2+ channel function is altered during pregnancy was tested by comparing responses to potassium chloride (KCl) and phenylephrine in aortic rings of virgin and term-pregnant rats under the influence of nifedipine and Bay K 8644. Maximum response to KCl was progressively reduced by increasing nifedipine concentrations (1.0-100 nM) in both groups of tissues. Nifedipine produced a smaller inhibition of KCl-induced contraction in aortic rings of pregnant than of virgin rats. It exerted little inhibition on the concentration response curve to phenylephrine. The Ca2+ channel antagonist (100 nM) reduced the maximum response to the alpha-adrenoceptor agonist in rings from virgin rats, but had no effect in pregnant rats. Bay K 8644, a Ca2+ channel activator, potentiated the responses to low concentrations of both phenylephrine and KCl in the tissues of both virgin and pregnant rats, but did not affect maximum responses. It also induced concentration-dependent contractions in rings of virgin but not of pregnant rats. The effects of Bay K 8644 were markedly potentiated by precontracting the aorta with 10mM KCl. Nevertheless tissues from pregnant rats were still less responsive to Bay K 8644. However, when the strips were precontracted to the same level by different concentrations of KCl, the concentration-response curves to Bay K 8644 were identical in both groups. [3H]Nitrendipine binding to membrane preparations of the thoracic aorta was similar in virgin and pregnant rats.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7498250 TI - Biochemical and behaviour changes induced by acute stress in a chronic variate stress model of depression: the effect of amitriptyline. AB - This paper examines the biochemical and behaviour changes induced by an acute stress (five 10-s, 1-mA foot-shocks) in three groups of rats: (1) never stressed, (2) subjected to chronic variate stress for 20 days, (3) subjected to the same chronic stress and treated with 5 mg/kg per day amitriptyline. After 15 min, acute stress led to a marked reduction in cortical beta-adrenoceptor and 5-HT2 receptor density, whereas the density of the 5-HT1A receptors was unchanged. Chronic stress also increased beta-adrenoceptor and 5-HT2 receptor density and had no effect on 5-HT1A. Acute stress diminished the density of beta adrenoceptors in chronically stressed animals, but did not alter that of the two 5-HT populations. Amitriptyline alone reduced beta-adrenoceptor and 5-HT2 receptor densities only. Acute stress applied to animals treated with amitriptyline reduced 5-HT1A receptors, and caused a further beta-adrenoceptor decrease, but had no further effect on the 5-HT2 receptors. On behaviour, chronic stress diminished reactivity to the acute stress. This reduction was fully abolished by amitriptyline. An open-field study showed that acute stress reduced motor activity, increased latency times and diminished rearing in the controls, whereas chronic stress reduced motor activity only. No significant changes in behaviour were induced by the acute stress in animals subjected to chronic stress. The combination of chronic stress with amitriptyline was accompanied by a diminution of exploratory activity that persisted after the acute stress.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7498249 TI - Angiotensin AT1 receptor-mediated attenuation of cardiac hypertrophy due to volume overload: involvement of endothelin. AB - The role of angiotensin II via the angiotensin type 1 (AT1) receptor in the development of volume overload cardiac hypertrophy was investigated in adult male Wistar rats with aortic insufficiency. We examined the effects of specific angiotensin AT1 receptor blockade with losartan (2-n-butyl-chloro-5-hydroxymethyl 1-[(2'-(1H-tetrazol-5-yl) biphenyl-4-yl)methyl]-imidazole potassium) on left ventricular weight and left ventricular angiotensin II and endothelin-1 level to test the possibility that the cardiac action of angiotensin II may be mediated by endogenous endothelin-1. Moreover, to verify the possible involvement of endothelin-1, we measured left ventricular endothelin-1 levels during the hypertrophic process and evaluated the effect of the endothelin ETA receptor specific antagonist, FR139317 ((R)2-[(R)2-[(S)-2-[[1-(hexahydro-1H azepinyl)]carbonly] amino-4-methylpentanoyl]amino-3-[3-(1-methyl-1H-indolyl) propionyl]amino-3-(2-pyridyl) propionic acid), on left ventricular weight. Two weeks after production of aortic insufficiency, left ventricular weight and left ventricular endothelin-1 concentration were markedly elevated in the rats with aortic insufficiency as compared with the sham-operated control rats, but left ventricular angiotensin II was not changed. Losartan (10 mg/kg/day p.o., 2 weeks) significantly reduced left ventricular weight and left ventricular endothelin-1 level in the rats with aortic insufficiency without affecting blood pressure and there was a significant positive correlation between left ventricular weight and left ventricular endothelin-1 content. Left ventricular endothelin-1 content correlatively increased to left ventricular weight during the development of the left ventricular hypertrophy.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7498251 TI - Involvement of nitric oxide in neuroglycopenia-induced insulin and glucagon secretion in the mouse. AB - Neuroglycopenia induced by administration of 2-deoxy-D-glucose is known to stimulate the secretion of both insulin and glucagon in mice by a mechanism that is dependent on neural activity. In the present study, we examined whether the neurotransmitter nitric oxide (NO) is involved in this process. Therefore, 2 deoxy-D-glucose (500 mg/kg) was injected intravenously alone or together with the inhibitor of NO synthase, NG-nitro-L-arginine methyl ester (50 mg/kg) to conscious mice. It was found that NG-nitro-L-arginine methyl ester inhibited the increased plasma levels of both insulin (by 26%; P = 0.039) and glucagon (by 45%; P < 0.001) at 10 min after injection of 2-deoxy-D-glucose. Similarly, the NO synthase inhibitor, NG-nitro-L-arginine, which is devoid of the anticholinergic property of NG-nitro-L-arginine methyl ester, inhibited the responses of both insulin (by 53%; P = 0.026) and glucagon (by 57%; P = 0.003) to 2-deoxy-D glucose. In contrast, the stereoisomer of NG-nitro-L-arginine methyl ester, NG nitro-D-arginine methyl ester, which is devoid of NO synthase inhibitory activity, was without effect on 2-deoxy-D-glucose-induced insulin and glucagon secretion. Plasma levels of adrenaline and noradrenaline after administration of 2-deoxy-D-glucose were also reduced by NG-nitro-L-arginine methyl ester. In contrast, the insulin and glucagon secretory responses to intravenous injection of arginine (250 mg/kg), glucose (500 mg/kg) or the cholinergic agonist, carbachol (30 micrograms/kg), were not influenced by NG-nitro-L-arginine methyl ester, NG-nitro-D-arginine methyl ester or NG-nitro-L-arginine.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7498252 TI - Behavioral and neurochemical actions of the strychnine-insensitive glycine receptor antagonist, 7-chlorokynurenate, in rats. AB - The present study investigated if blockade of the modulatory glycine receptor of the NMDA receptor complex influences the expression of behavior (sniffing stereotypy and locomotion) and dopamine metabolism in rats as it has been shown for NMDA receptor antagonists. The glycine receptor antagonist, 7 chlorokynurenate (7-chloro-4-hydroxyquinoline-2-carboxylic acid), induced a dose dependent sniffing stereotypy but had no effect on locomotion when it was given i.c.v. The glycine receptor agonist, D-cycloserine (D-4-amino-3-isoxazolidinone), antagonized the sniffing stereotypy. 7-Chlorokynurenate had no influence on dopamine metabolism in the striatum and the nucleus accumbens, but moderately decreased the metabolism in the prefrontal cortex. Comparison of behavioral and neurochemical outcomes suggests that the failure to induce locomotion correlates with the unchanged dopamine metabolism in the basal ganglia, while sniffing stereotypy does not. These results show that blockade of the glycine receptor of the NMDA receptor complex induces a behavioral and neurochemical profile similar to that of competitive NMDA receptor antagonists. PMID- 7498253 TI - Protective effect of riluzole on excitatory amino acid-mediated neurotoxicity in motoneuron-enriched cultures. AB - Excitatory amino acid-mediated neurotoxicity was investigated in motoneuron enriched cultures from fetal rats at 12-14 days of gestation. The cultures were mainly composed of differentiated motoneurons identified by choline acetyl transferase and calcitonin gene-related peptide (CGRP) immunoreactivity. Addition of glutamate (600 microM) to the conditioned medium induced no acute neuronal swelling. However, it was followed by a widespread neuronal degeneration over the next 24 h, accounting for 77% of the total cell number. Glutamate toxicity was dose dependent, with an EC50 around 300 microM. Treatment for 24 h with the agonists, N-methyl-D-aspartate (NMDA, 100 microM), kainate (500 microM) or RS alpha-amino-3-hydroxy-5-methyl-4-isoxalopropionate (AMPA, 10 microM), also induced a significant cell loss. Riluzole (2 amino 6 trifluoromethoxybenzothiazole), a compound known to interfere with glutamatergic transmission pre- and postsynaptically, significantly reduced glutamate and NMDA neurotoxicity in a dose-dependent manner. These results suggest that a prolonged activation of one or more subtypes of ionotropic excitatory amino acid receptors can lead to motoneuron degeneration in vitro, and provide direct experimental evidence supporting the neuroprotective effect of riluzole in cultured motoneurons. PMID- 7498255 TI - Reduction of gastrointestinal injury in acute endotoxic shock by flurbiprofen nitroxybutylester. AB - Nitric oxide has been reported to have paradoxical effects in experimental endotoxic shock, contributing to the hemodynamic consequences of endotoxin administration, but apparently protecting the gastrointestinal mucosa. A novel class of nitric oxide-releasing nonsteroidal anti-inflammatory drug (NSAID) derivatives has recently been described which exert anti-inflammatory activities but produce significantly less gastrointestinal injury than the parent nonsteroidal anti-inflammatory drugs from which they are derived. Thus, the present study was performed to determine the effects of one of these derivatives, flurbiprofen 4-nitroxybutylester, compared to the native nonsteroidal anti inflammatory drug, flurbiprofen, in an experimental model of endotoxic shock. Intravenous administration of endotoxin from Salmonella typhosa to rats pretreated with flurbiprofen produced a profound decrease in systemic arterial blood pressure, an increase in hematocrit and extensive gastric and small intestinal damage. In rats pretreated with flurbiprofen 4-nitroxybutylester, endotoxin produced comparable changes in blood pressure and hematocrit to those seen in rats treated with flurbiprofen; however, the severity of gastrointestinal damage was significantly reduced. Gastric blood flow was profoundly decreased following endotoxin administration, but was significantly higher in rats pretreated with flurbiprofen 4-nitroxybutylester than in rats pretreated with flurbiprofen. These results demonstrate that despite not affecting the acute systemic effects of endotoxin administration, flurbiprofen 4-nitroxybutylester is capable of protecting the gastrointestinal mucosa from injury, possibly through preservation of mucosal blood flow. PMID- 7498256 TI - Endothelium-dependent relaxation of rat aorta by butein, a novel cyclic AMP specific phosphodiesterase inhibitor. AB - Butein, isolated from Dalbergia odorifera T. Chen, caused endothelium-dependent relaxation of rat aorta precontracted with phenylephrine. This effect was abolished in endothelium-denuded aorta and in endothelium-intact aorta in the presence of NG-monomethyl-L-arginine, oxyhemoglobin and methylene blue, whereas the effect was unaltered by indomethacin or charybdotoxin. These results indicate that the vasorelaxant effect of butein depended on the endothelium and was mediated by endothelium-derived relaxing factor (EDRF). Incubation of endothelium intact aorta with butein increased not only cAMP but also cGMP content. Four phosphodiesterase forms were isolated by diethylaminoethyl (DEAE)-Sephacel chromatography from rat aorta. cAMP-specific phosphodiesterase (type IV) activity was potently inhibited by butein with an IC50 of 10.4 +/- 0.4 microM. In contrast, phosphodiesterase I, III and V activities were inhibited by butein above 100 microM. Adenylate cyclase and guanylate cyclase activities were unchanged by butein. These results suggest that the increase of cAMP formation elicited by butein is due to the inhibition of cAMP-specific phosphodiesterase. The specific phosphodiesterase IV inhibitor (rolipram) and V inhibitor (zaprinast) both produced endothelium-dependent relaxations, whereas the phosphodiesterase III inhibitor (trequinsin) produced relaxation of rat aorta independent of the endothelium. In the presence of a functional endothelium, relaxations produced by butein were significantly potentiated by isoprenaline, forskolin, trequinsin and sodium nitroprusside. It is concluded that butein, a novel cAMP-specific phosphodiesterase inhibitor, produced relaxation of rat aorta, an effect dependent on an intact endothelium. The relaxant effect of butein was markedly enhanced by cGMP-elevating agents.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7498254 TI - The delta-opioid receptor antagonist naltrindole prevents sensitization to the conditioned rewarding effects of cocaine. AB - A conditioned place preference paradigm was used to determine whether: (i) prior exposure to cocaine results in an enhancement of its rewarding effects, and (ii) the delta-opioid receptor antagonist naltrindole can prevent the development of this response. Rats received daily injections of saline or cocaine (10.0 mg/kg i.p.) for 5 days in the colony room. Additional animals received naltrindole (0.03-0.3 mg/kg s.c.), lithium chloride (20 mg/kg s.c.) or vehicle prior to i.p. injections. Conditioning sessions (2 drug; 2 vehicle) commenced 3 days later. Cocaine (1.0-10.0 mg/kg) was ineffective as a conditioning stimulus in saline pre exposed rats. In cocaine pre-exposed animals, however, doses of 5.0 and 10.0 mg/kg cocaine resulted in significant drug-induced place preferences. Significant cocaine-induced place preferences were also observed in animals which had received lithium chloride with the cocaine treatment regimen. In animals which had received naltrindole together with the chronic cocaine treatment regimen, cocaine failed to produce a conditioned response. These data demonstrate that the repeated administration of cocaine results in an enhancement of its rewarding effects (e.g. sensitization) and that this phenomenon is prevented by a delta opioid receptor antagonist. Furthermore, the finding that naltrindole does not modify the acute rewarding effects of cocaine suggests a specific role of delta opioid receptors in the sensitization process. PMID- 7498257 TI - Erysodine, a competitive antagonist at neuronal nicotinic acetylcholine receptors. AB - Erysodine, an erythrina alkaloid related to dihydro-beta-erythroidine, was found to be a more potent inhibitor of [3H]cytisine binding at neuronal nicotinic acetylcholine receptors but a less potent inhibitor of [125I]alpha-bungarotoxin binding at muscle-type nicotinic acetylcholine receptors than dihydro-beta erythroidine. Erysodine was a competitive, reversible antagonist of (-)-nicotine induced dopamine release from striatal slices and inhibited (-)-nicotine-induced 86Rb+ efflux from IMR-32 cells. Erysodine was equipotent with dihydro-beta erythroidine in the dopamine release assay but 10-fold more potent in the 86Rb+ efflux assay, suggesting differential subtype selectivity for these two antagonists. Erysodine, systemically administered to mice, entered the brain and significantly attentuated nicotine's hypothermic effects and its anxiolytic-like effects in the elevated plus-maze test. There was greater separation between antagonist and toxic doses for erysodine than for dihydro-beta-erythroidine, perhaps because of erysodine's greater selectivity for neuronal receptors. In rats, erysodine prevented both the early developing decrease and the late developing increase in locomotor activity produced by (-)-nicotine. The potent and competitive nature of erysodine's antagonism together with its ability to enter the brain after systemic administration suggest that erysodine may be a useful tool in characterizing neuronal nicotinic acetylcholine receptors. PMID- 7498258 TI - Adrenomedullin and calcitonin gene-related peptide in the rat isolated kidney and in the anaesthetised rat: in vitro and in vivo effects. AB - The effects of adrenomedullin and calcitonin gene-related peptide (CGRP) were compared in the rat isolated perfused kidney and in the anaesthetised rat. Adrenomedullin and CGRP both elicited concentration-dependent vasodilator responses from perfused kidneys (adrenomedullin was less potent than CGRP). These responses were blocked by CGRP-(8-37) (1 microM). Adrenomedullin and CGRP elicited dose-dependent hypotension in anaesthetised rats (adrenomedullin was less potent than CGRP). The hypotensive responses to CGRP, but not those to adrenomedullin, were antagonized by CGRP-(8-37) (12 nmol/kg/min). These studies indicate that the in vivo response to adrenomedullin may be mediated through CGRP (8-37) insensitive receptors. PMID- 7498261 TI - The preferential dopamine D3 receptor ligand, (+)-UH232, is a partial agonist. AB - In a NG 108 15 hybrid cell line stably expressing a recombinant dopamine D3 receptor, (+)-UH 232 (cis-(+)-1S,2R)-5-methoxy-1-methyl-2-(di-n- propylamino)tetralin), a partially selective D3 receptor ligand, stimulates mitogenesis, as measured by incorporation of [3H]thymidine, with an EC50 of 7.6 nM and a maximal increase corresponding to 23% of the response elicited by quinpirole, a full agonist. This effect was antagonised by nafadotride, a D3 receptor-selective antagonist. (+)-UH 232 also antagonised quinpirole-induced mitogenesis with a Ki value of 9.4 nM. (+)-UH 232 (1 microM) inhibited by 22% the forskolin-induced accumulation of cAMP, whilst the inhibition by quinpirole (100 nM) was 53%. These results indicate that (+)-UH 232 is a partial agonist at the D3 receptor with an intrinsic activity of 0.2-0.4. PMID- 7498259 TI - K+ channel blockers do not modify relaxation of guinea-pig uterine artery evoked by acetylcholine. AB - The effect of K+ channel blockers on acetylcholine-induced relaxation in guinea pig uterine arterial rings was investigated. Acetylcholine (0.1 nM-60 microM) induced endothelium-dependent relaxation of phenylephrine-precontracted guinea pig uterine artery. Methylene blue (30 nM-1 microM) and NG-monomethyl-L-arginine (3-30 microM) antagonized the effect of acetylcholine, with suppression of the maximal acetylcholine-induced relaxation, in a concentration-dependent manner. The inhibition of relaxation by NG-monomethyl-L-arginine (10 microM) was significantly overcome by L-arginine (10 microM), but not by D-arginine (100 microM). In contrast, the administration of K+ channel blockers, tetraethylammonium (6 mM), glibenclamide (5 microM), apamin (1 microM) and 4 aminopyridine (1 mM), did not modify the relaxation of guinea-pig uterine artery induced by acetylcholine. The concomitant addition of K+ channel blockers in the same concentrations also did not alter the inhibition of acetylcholine-induced relaxation produced by NG-monomethyl-L-arginine (30 microM). Finally, the acetylcholine-evoked relaxations were unaltered when K(+)-rich Krebs-Ringer bicarbonate solution was used to induce tone instead of phenylephrine. Indomethacin (10 microM) and diethylcarbamazine (100 microM) had no effects on acetylcholine-induced relaxation. These findings indicate that K+ channels are probably not involved in the endothelium-dependent guinea-pig uterine arterial relaxation elicited by acetylcholine. PMID- 7498260 TI - Cannabinoid precipitated withdrawal by the selective cannabinoid receptor antagonist, SR 141716A. AB - Precipitated withdrawal in rats chronically exposed to delta 9 tetrahydrocannabinol, the major psychoactive principle of the marijuana plant, was unequivocally demonstrated for the first time using a selective antagonist, SR 141716A (N-(piperidin-1-yl)-5-(4-chlorophenyl)-1(2,4- dichloro-phenyl)-4 methyl-1H-pyrazole carboxamide.HCl). This demonstration should provide a powerful stimulus for the systematic study of dependency on the psychoactive cannabinoids. PMID- 7498262 TI - Effect of nitrendipine on renal cortical and papillary autoregulation in hypertensive rats. AB - Renal cortical and papillary perfusions were assessed by laser-Doppler flowmetry and autoregulatory indices. There was cortical autoregulation (autoregulatory index of 0.31 +/- 0.08) in Wistar from 160 to 100 mm Hg, but not in the papilla, which was abolished by nitrendipine, 0.125 and 0.25 micrograms kg-1 min-1. In SHRSP the cortex, but not the papilla, exhibited autoregulation from 180 to 120 mm Hg (autoregulatory index of 0.27 +/- 0.10) but not during low and high doses of nitrendipine. The non-clipped kidney cortex of 2K1C Goldblatt rats autoregulated from 190 to 130 mm Hg (autoregulatory index of 0.20 +/- 0.13), but was inhibited only by the higher dose of nitrendipine. In mature Wistar and SHRSP cortical autoregulation is blocked effectively by nitrendipine whereas the 2K1C Goldblatt hypertensive rats are relatively resistant. PMID- 7498263 TI - LCB 2183 inhibits tracheal hyperreactivity and pulmonary inflammation in mouse airways. AB - The pulmonary delayed-type hypersensitivity (DTH) reaction induced by picryl chloride is characterized by enhanced albumin concentration in the airway tissue in the early phase (2 h after challenge with picryl sulphonic acid). During the later phase (48 h after the challenge) enhanced tracheal reactivity to carbachol in vitro and cellular (mononuclear and polymorphonuclear leukocytes) accumulation into the airway tissue in vivo are prominent features. In this present study, the effects of a novel drug, LCB 2183, were examined in the pulmonary DTH reaction. LCB 2183 (25 mg/kg twice daily intragastric gavage) failed to inhibit the enhanced albumin accumulation in the early phase of this response. In contrast, LCB 2183 abolished the tracheal hyperreactivity and the leukocyte accumulation in the airways of picryl chloride-sensitized mice 48 h after the challenge. These results demonstrate that LCB 2183 could be effective in the treatment of airway hyperreactivity and pulmonary inflammation. PMID- 7498264 TI - The effect of urethane anesthesia on evoked potentials in dentate gyrus. AB - We examined the effect of urethane (1000 mg/kg, followed by 50 mg/kg per h, i.v.), an anesthetic commonly used by physiologists, on evoked potentials recorded in dentate gyrus in adult Wistar rats by stimulating the ipsilateral perforant path, via chronically implanted electrodes. Urethane decreased paired pulse inhibition. Under urethane, with paired-pulse stimulation, the ratio of the second population spike amplitude to the first increased by 11.1-20.7% at 25-60 ms interstimulus interval (n = 18, P < 0.05). At 25 ms, the proportion was 3.6 +/ 1.6 while awake, and 14.7 +/- 3.5 under urethane. Urethane depressed granule cell excitability and strength of synaptic responses. Under urethane, the ratio of the population spike amplitude obtained at 250 microA stimulation to the maximal response in the same input/output response examination decreased by 20%, and the ratio of the excitatory postsynaptic response slopes fell by 10%. These results indicate that urethane affects neurotransmission in the hippocampus, and suggest that its effect may be exerted in part on excitatory neurotransmission. PMID- 7498265 TI - Capsaicin pre-treatment prevents the development of antigen-induced airway hyperresponsiveness in neonatally immunised rabbits. AB - The effect of a 3-day pre-treatment regime of capsaicin (8-methyl-N-vanillyl-6 nonenamide) (80 mg/kg s.c.) on airway changes induced by Alternaria tenuis aerosol challenge 3 days later was assessed in adult rabbits immunised from birth to the age of 3 months. Pre-treatment with capsaicin did not alter basal lung function or basal responsiveness to inhaled histamine. While capsaicin had no significant effect on the acute bronchoconstriction induced by antigen, this dose was sufficient to significantly inhibit the increase in airway responsiveness to inhaled histamine achieved 24 h following antigen challenge. The pulmonary recruitment of neutrophils and eosinophils induced by antigen was unaltered by prior treatment with capsaicin. In vitro contractile responsiveness to methacholine was not significantly different in bronchial tissues removed from capsaicin- and vehicle-pre-treated rabbits. In addition, there were no significant differences in responses to methacholine in preparations denuded of epithelium. Contraction of bronchial tissue induced by exogenously applied capsaicin in vitro, although modest, was significantly inhibited in capsaicin-pre treated animals. In vehicle-pre-treated rabbits, contraction induced by a second challenge with capsaicin 45 min later was significantly reduced to a level that made responses not significantly different from those obtained in capsaicin-pre treated tissues. The results of the present study demonstrate that antigen induced airway hyperresponsiveness to inhaled histamine in immunised rabbits is inhibited by prior treatment with capsaicin. These findings suggest the involvement of capsaicin-sensitive nerves in antigen-induced airway hyperresponsiveness but not acute bronchospasm or cell infiltration induced by antigen. PMID- 7498267 TI - Studies of four novel diphenylbutylpiperazinepyridyl derivatives on release and inhibition of reuptake of dopamine, serotonin and noradrenaline by rat brain in vitro. AB - Four novel diphenylbutylpiperazinepyridyl derivatives (FG5865 (2-[4-[4,4-bis(4 fluorophenyl)butyl]-1-piperazinyl]-3-pyridine- carboxamide), FG5891 (2-[4-[4,4 bis(4-fluorophenyl)butyl]-1-piperazinyl]-N-methyl-3- pyridinecarboxamide), FG5893 (2-[4-[4,4-bis(4-fluorophenyl)butyl]-1-piperazinyl]-3- pyridinecarboxylic acid methyl ester) and FG5909 (2-[4-[4,4-bis(4-fluorophenyl)butyl]-1-piperazinyl]-3 hydroxypyridine) were tested concerning their effects on in vitro release and reuptake of neurotransmitters. Serotonin, noradrenaline and dopamine were those considered, with rat synaptosomes prepared respectively from frontal cortex, cortex and striatum. FG5865, FG5891, FG5893 and FG5909 were found to potently inhibit the uptake of all three neurotransmitters. In addition, FG5865, FG5891 and FG5893 increased the release of serotonin and dopamine from perfused frontal cortical and striatal tissue; FG5865 was most potent in this regard. The release induced by the FG compounds was, however, much less than that induced by e.g. fenfluramine or amphetamine. All four FG compounds were also found to inhibit glutamate-stimulated release of dopamine from striatal tissue; for FG5893 this inhibition occurred at nanomolar concentrations. PMID- 7498266 TI - Different effects of isoproterenol and dihydroouabain on cardiac Ca2+ transients. AB - Cytosolic fura-2 Ca2+ transient signals (TCa) and the left ventricular pressure or contraction of myocardium under the positive inotropic effects of the beta adrenoceptor agonist, isoproterenol, and the cardiac glycoside, dihydroouabain, were measured simultaneously and the results were compared. TCa was observed preceding the onset of force development and showed a steeper rise and slower decay than did the contraction curve of papillary muscle. Isoproterenol increased the steepness and the amplitude of TCa, reflecting the speed and peak force of contraction, and clearly biphasic TCa were observed with biphasic contractions developed at low frequency. Ryanodine reduced not only the early component of the contraction but also TCa, without affecting the diastolic Ca2+ level. These effects of isoproterenol were attributed to the enhanced uptake of Ca2+ by the sarcoplasmic reticulum. In contrast, dihydroouabain elevated the Ca2+ level at diastole without any change in the amplitude of TCa, suggesting that dihydroouabain inhibits the membrane Na pump thereby increasing the intracellular Ca2+ via Na(+)-Ca2+ exchange. Furthermore, a comparison of the time course of the isometric twitch curve with that of TCa in rested state contraction indicated that there are distinct differences between the mechanisms of the positive inotropic effects of isoproterenol and of dihydroouabain. PMID- 7498268 TI - Why does clozapine stimulate the motor activity of reserpine-pretreated rats when combined with a dopamine D1 receptor agonist? AB - The aim of the present experiments was to investigate the locomotor stimulant effects of the atypical antipsychotic agent, clozapine, in rats depleted of their dopamine by reserpine and alpha-methyl-p-tyrosine pretreatment. Clozapine itself induced a slight but never significant stimulation of locomotor activity which was enhanced by the addition of the selective dopamine D1 receptor agonist, SKF38393 (2,3,4,5-tetrahydro-7,8-dihydroxy-1-phenyl-1H-3- benzazepine), but not by the selective dopamine D2 receptor agonist, quinpirole. The stimulation produced by clozapine plus SKF38393 was blocked by the selective dopamine D1 receptor antagonist, SCH23390 (7-chloro-8-hydroxy-3-methyl-1-phenyl-2,3,4,5- tetrahydro-1H-3-benzapine hydrochloride), while the selective dopamine D2 receptor antagonist, haloperidol, was ineffective. A combination of SCH23390 and haloperidol blocked the clozapine plus SKF38393-induced locomotion. Unlike clozapine, neither the selective 5-HT2 receptor antagonist, ritanserin, nor the dopamine D2 receptor antagonists, haloperidol and remoxipride, caused locomotor activation when given alone or in combination with SKF38393. The indirectly acting sympathomimetic amine, d-amphetamine, was inactive in the monoamine depleted rats, indicating that no dopamine was available for release by d amphetamine. The muscarinic receptor antagonist, scopolamine, alone did not alter locomotion, but produced marked stimulation when combined with SKF38393 but not with quinpirole. This stimulation was not affected by haloperidol. However, the scopolamine plus SKF38393-induced stimulation was partially blocked by SCH23390 or by a combination of haloperidol and SCH23390. The data indicate that clozapine, in rats depleted of their dopamine stores, exhibits properties consistent with those of a dopamine receptor agonist.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7498269 TI - Interaction of 5-aminosalicylic acid with nitric oxide on rat aortic strips and human platelets. AB - We have examined the interactions of 5-aminosalicylic acid with nitric oxide (NO). Phenylephrine-precontracted rat aortic strips with intact endothelium were further contracted by 5-aminosalicylic acid (50-200 microM) in a concentration dependent manner. Removal of endothelium, inhibition of guanylate cyclase by methylene blue, inhibition of NO biosynthesis by NG-nitro-L-arginine as well as in inactivation of NO by oxyhemoglobin abolished the effect of 5-aminosalicylic acid. The antiaggregatory effects of 3-morpholinosydnonimine and rat peritoneal neutrophils, which are due to release of NO, were diminished in a concentration dependent manner by 5-aminosalicylic acid (50-250 microM). In both experimental models the effects of 5-aminosalicylic acid were significantly reduced by superoxide dismutase in a concentration which alone exhibited no effect. Since NO might act as a cytotoxic and vasodilating mediator, our results suggest that inactivation of NO by 5-aminosalicylic acid could contribute to the therapeutic activity of the drug in inflammatory bowel disease. PMID- 7498270 TI - Platelet-activating factor-induced loss of vascular responsiveness to noradrenaline in pithed rats: involvement of nitric oxide. AB - The role of nitric oxide and cyclo-oxygenase products in the platelet-activating factor (PAF)-induced hyporesponsiveness to noradrenaline was investigated in pithed rats. Infusion of PAF (30 ng/kg/min) for 60 min reduced the mean arterial blood pressure and impaired the pressor responses to noradrenaline (10 ng/kg, 100 ng/kg, 1 microgram/kg). Administration of NG-monomethyl-L-arginine (L-NMMA, 30 mg/kg) restored the reduced MABP and the impaired responses to their original levels. Indomethacin (5 mg/kg) had no significant effect on the PAF-induced hyporesponsiveness. Administration of 30 mg/kg L-NMMA caused hypertension in the PAF vehicle-treated animals and reduced the pressor response to 1 microgram/kg noradrenaline. Administration of 3 mg/kg L-NMMA had no significant effect on the responsiveness to noradrenaline. These results suggest that nitric oxide contributes to the PAF-induced hyporesponsiveness to noradrenaline and that cyclo oxygenase products do not play a major role in this hyporesponsiveness. PMID- 7498271 TI - Possible involvement of medial prefrontal cortex in amphetamine-induced sensitization of mesolimbic dopamine function. AB - We examined the role of the dopamine projection to the medial prefrontal cortex in amphetamine-induced sensitization of meso-nucleus accumbens dopamine function. In the first experiment, male rats received bilateral microinfusions either of 6 hydroxydopamine or of vehicle (sham) into prefrontal cortex. Six weeks later animals from both groups were injected once daily for 5 consecutive days with either amphetamine or saline. Two days after the last daily injection, all the animals were each implanted with a voltammetric electrode into nucleus accumbens. Increases in dopamine-dependent electrochemical signals elicited by amphetamine were monitored 3-4 days later using chronoamperometry. The results showed that amphetamine stimulates dopamine efflux to a greater extent in the nucleus accumbens of lesioned than of sham-lesioned animals. Furthermore, of the animals with prefrontal cortical lesions, amphetamine-induced dopamine efflux was greater in animals previously treated with the drug than in animals with no prior drug experience. In a second experiment, sensitization to the acute locomotor stimulant effect of amphetamine was examined in prefrontal cortex-lesioned and sham-lesioned animals. The locomotor response of all animals to a test dose of amphetamine was first monitored and then on each of the subsequent 5 days, lesioned and sham-lesioned animals received an injection either of amphetamine or of saline. Five and then 13 days later, the locomotor response of all animals to the test dose of amphetamine was again measured. The results of this study showed that prefrontal cortex-lesioned animals were less responsive to the first amphetamine injection than sham-lesioned animals. However, after repeated daily administration, the acute locomotor response of lesioned animals to amphetamine was significantly greater than that of sham-lesioned animals with the same drug history. These findings are generally consistent with evidence from other sources suggesting that the dopamine input to medial prefrontal cortex exerts an indirect, inhibitory influence on mesolimbic dopamine transmission. They also suggest that long-term changes to a dopamine-sensitive mechanism in prefrontal cortex may contribute to the development of stimulant-induced sensitization of mesolimbic dopamine function. PMID- 7498273 TI - Mechanisms involved in the relaxant response of bradykinin in epithelium intact strips of the guinea-pig trachea. AB - Kinins caused graded relaxations in guinea-pig trachea with epithelium under spontaneous or carbachol-induced tone. The order of potency was: [Tyr8]bradykinin > lysyl-bradykinin > bradykinin > methionyl-lysyl- bradykinin. The bradykinin B1 receptor agonist des-Arg9-bradykinin (1 microM) was inactive. Relaxation in response to bradykinin (100 nM) was unaffected by tetrodotoxin (0.3 microM), nicardipine (1 microM), Ca(2+)-free solution without or plus ryanodine (10 microM), propranolol (1 microM), glibenclamide (1 microM), staurosporine (0.3 microM), nickel chloride (100 microM) or [D-p-Cl-Phe6,Leu17]VIP (a vasoactive intestinal peptide receptor antagonist, 0.03 microM), but was partially inhibited by apamin (0.3-1 microM). Both HOE 140 (D-Arg-[Hyp3,Thi5,D-Tic7,Oic8]bradykinin) and NPC 17761 (D-Arg0[Hyp3,D-HypeE(trans-thiophenyl)7,Oic8]bra dykinin) (0.1-1000 nM) caused graded, reversible and selective inhibition of the bradykinin (100 nM) relaxation, with IC50 values of 1.4 and 19.1 nM, respectively. HOE 140 and NPC 17761 (0.1-10 nM) produced a graded shift to the right of the bradykinin concentration-response curves associated with a reduction of the maximum relaxation. The kinin B1 receptor antagonist, des-Arg9-[Leu8]bradykinin (1 microM), was inactive. Thus, bradykinin-induced relaxation in guinea-pig trachea results from activation of bradykinin B2 receptors and can be antagonized with high affinity in a selective and reversible manner, through noncompetitive mechanism, by both HOE 140 and NPC 17761. In addition, the bradykinin response does not involve neural pathways, extracellular Ca2+ influx or mobilization of intracellular Ca2+ stores sensitive to ryanodine, but is modulated by small conductance Ca(2+)-activated K+ channels. PMID- 7498272 TI - Diazepam potentiates the positive inotropic effect of isoprenaline in rat ventricle strips: role of cyclic AMP. AB - The responses of the electrically driven right ventricle strip of the rat heart to isoprenaline and other cyclic AMP-related inotropic agents were recorded in the absence and in the presence of diazepam. Isoprenaline, in concentrations ranging from 10 nM to 1 microM, significantly increased, in a concentration dependent manner, the contractile force in this preparation. Diazepam (10 microM) produced a leftward shift in the isoprenaline concentration-response curve and significantly reduced its EC50. Higher concentrations of diazepam (100 microM) produced no further shift, but reduced the maximum of the concentration-response curve of isoprenaline. Forskolin (0.5-10 microM), which directly stimulates adenyl cyclase, also produced a concentration-dependent increase in cardiac contractility. Diazepam (10 microM) displaced to the left the concentration response curve for forskolin and reduced its EC50. The cyclic AMP analogous dibutyryl cyclic AMP (0.1-1 mM) produced concentration-dependent positive inotropic effects which were not significantly modified in the presence of diazepam (10 microM). Diazepam (10 microM) significantly enhanced the cyclic AMP production induced by isoprenaline (0.1 microM) and forskolin (10 microM) by about 136% and 35% respectively. These results indicate that diazepam potentiates the positive inotropic effect induced by beta-adrenoceptor agonists, probably by increasing cyclic AMP production induced by these agents. PMID- 7498274 TI - Effects of dynorphin A-(1-13) on carbon monoxide-induced delayed amnesia in mice studied in a step-down type passive avoidance task. AB - The effects of dynorphin A-(1-13) on carbon monoxide (CO)-induced amnesia in mice were investigated using a step-down type passive avoidance task. Memory deficiency occurred in mice when training commenced 7 days after CO exposure although it was not produced 1 day after CO exposure. The median step-down latency in the retention test of the CO-exposed group was significantly shorter than that of the control group. Administration of dynorphin A-(1-13) (1.5 nmol/mouse i.c.v.) 15 min before the first training session prolonged the step down latency in the CO-exposed group. Dynorphin A-(1-13) administered immediately after the first training session or administered 15 min before the retention test also prolonged the step-down latency in the CO-exposed group. To determine whether this effect of dynorphin A-(1-13) was mediated via kappa-opioid receptors, we attempted to block its action using a kappa-opioid receptor antagonist (nor-binaltorphimine). Nor-binaltorphimine (5.44 nmol/mouse i.c.v.) blocked the effect of dynorphin A-(1-13) on delayed amnesia. However, dynorphin A (1-13) (0.5, 1.5 and 5.0 nmol/mouse) did not facilitate the acquisition of memory in normal mice. These results suggest that dynorphin A-(1-13) modulates the kappa opioid receptor-mediated opioid neuronal system, and that it ameliorates the disruptive effect of CO on acquisition, consolidation and/or recall of memory. PMID- 7498275 TI - Involvement of alpha 2-adrenoceptors in the cardiovascular effects of moxonidine. AB - The central sympathoinhibition caused by moxonidine has been explained by activation of alpha 2-adrenoceptors on the one hand, and by an action at imidazoline I1 receptors on the other hand. In order to examine these possibilities, effects of moxonidine were compared with those of 5-bromo-6-(2 imidazolin-2-ylamino)-quinoxaline (UK 14304), an alpha 2-adrenoceptor agonist with very low affinity for I1 receptors, in conscious rabbits. The interaction with yohimbine, an alpha 2-adrenoceptor antagonist with very low affinity for imidazoline I1 receptors, was also studied. Moxonidine 3-100 micrograms kg-1 and UK 14304 1-30 micrograms kg-1 i.v. elicited similar effects: they decreased arterial blood pressure after a transient increase, decreased renal sympathetic nerve activity (recorded with chronically implanted electrodes), decreased heart rate and decreased the plasma noradrenaline concentration. Yohimbine given i.v. antagonized the effects of moxonidine and of UK 14304 in a similar manner. Yohimbine injected into the cisterna magna (i.c.) prevented the hypotension but did not change the decrease in plasma noradrenaline and heart rate, again in the case of both moxonidine and UK 14304. The agreement of the effect patterns of moxonidine and UK 14304, and the similar antagonism of yohimbine against either drug, demonstrate involvement of alpha 2-adrenoceptors in their central sympathoinhibitory action. The resistance of the bradycardia and the plasma noradrenaline fall against yohimbine i.c. indicates a contribution of presynaptic alpha 2-adrenergic inhibition of transmitter release from postganglionic sympathetic neurons to the overall reduction of sympathetic tone. PMID- 7498276 TI - K+ channel openers produce epithelium-dependent relaxation of the guinea-pig trachea. AB - The relaxant effects of the K+ channel openers, NIP-121, (+)-7,8-dihydro-6,6 dimethyl-7-hydroxy-8-(2-oxo-piperidin-1-yl)-6H - pyrano[2,3-f]benz-2,1,3 oxadiazole, and cromakalim, were investigated in epithelium-intact and -denuded tracheal spirals isolated from guinea-pigs. In the presence of 5 microM indomethacin, NIP-121 (0.01-1 microM) and cromakalim (0.1-10 microM) relaxed, in a concentration-dependent manner, epithelium-intact and -denuded trachea precontracted with a thromboxane A2 mimetic, U46619, 9,11-dideoxy-9 alpha, 11 alpha-methanoepoxy-prostaglandin F2 alpha (30 nM). The relaxations of epithelium denuded trachea were significantly decreased as compared with those of epithelium intact trachea. The relaxations induced by salbutamol or aminophylline were not affected by epithelium removal. In epithelium-intact trachea, the NIP-121- and cromakalim-induced relaxations were not modulated by the neutral endopeptidase inhibitor, phosphoramidon (10 microM), or the nitric oxide synthesis inhibitor, N omega-nitro-L-arginine (100 microM). However, the guanylate cyclase inhibitor, methylene blue (100 microM), significantly reduced NIP-121- and cromakalim induced relaxation of epithelium-intact trachea. Methylene blue also reduced sodium nitroprusside-induced relaxation but did not affect isoprenaline-induced relaxation. These findings suggest that the K+ channel openers, NIP-121 and cromakalim, may induce, at least in part, epithelium-dependent and methylene blue sensitive relaxation of the guinea-pig isolated trachea. PMID- 7498277 TI - Functional analyses of alpha 1-adrenoceptor subtypes in rat hypothalamic ventromedial nucleus neurons. AB - Activation of alpha 1-adrenoceptors in rat hypothalamic ventromedial nucleus can excite neurons and facilitate female sexual behavior. To identify the alpha 1 adrenoceptor subtype(s) involved, the alpha 1B-adrenoceptor-specific antagonist chloroethylclonidine (100 microM) and/or the alpha 1A-adrenoceptor-selective antagonist 5-methyl urapidil (1 or 2.5 microM) or WB-4101 (0.1-10 microM) were applied to a recording chamber bathing the hypothalamic slice containing the ventromedial nucleus. In all the neurons tested, both types of antagonists blocked, often completely, excitatory responses to nonselective alpha 1 adrenoceptor agonists. Since the doses used were unlikely to make these antagonists nonselective, the results suggest that activation of both alpha 1A- and alpha 1B-adrenoceptor subtypes was necessary for alpha 1-adrenoceptor agonists to evoke an excitation, or that with the present application method- injection into the continuously perfused chamber--chloroethylclonidine did not act specifically. In preincubation (at 37 degrees C for 90 min) where it was reported to act by specific alkylation, chloroethylclonidine (100 microM) but not the vehicle abolished the excitation evoked by an alpha 1-adrenoceptor agonist, but not that by carbachol or other excitants. Also, either in bath application or incubation, chloroethylclonidine worked equally efficiently on slices from ovariectomized rats, that reportedly contain few alpha 1B-adrenoceptors, and from those treated with estrogen which induces alpha 1B-adrenoceptors selectively, suggesting that alpha 1B-adrenoceptor was necessary even when in low abundance.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7498278 TI - Delayed anti-inflammatory action of nedocromil sodium in the rat paw is dependent on de novo protein synthesis. AB - Nedocromil sodium is commonly suggested to reduce allergic inflammation by inhibiting mediator release from mast cells. However, nedocromil also exhibits a wide range of additional anti-inflammatory activities, including inhibition of increased vascular permeability induced by individual mediators such as histamine. In the present study, we have further characterized the mode of action of nedocromil in a rat model for hind paw edema. Mast cell-dependent edema was induced with compound 48/80 (edema response mainly due to 5-hydroxytryptamine release), and direct mediator-induced plasma extravasation was evoked by exogenous 5-hydroxytryptamine (both agents injected locally). Local pretreatment with nedocromil for 20 min dose-dependently inhibited the edema evoked by compound 48/80 more effectively than that induced by 5-hydroxytryptamine. However, after 2 h pretreatment, both the 5-hydroxytryptamine-and compound 48/80 induced edema responses were inhibited to approximately the same extent by a range of concentrations of nedocromil, as well as by dexamethasone. Local inhibition of RNA/protein synthesis with actinomycin-D abolished the effects of both dexamethasone and nedocromil (2 h local pretreatment). We thus conclude that nedocromil can produce an 'anti-exudative' effect that is independent of inhibition of mast cell mediator release, is slow in onset, and requires de novo protein synthesis. PMID- 7498279 TI - Possible neuronal origin of ATP release evoked by forskolin and ouabain from guinea-pig atrial segments. AB - The characteristics of ATP release evoked by forskolin and ouabain from atrial segments of guinea-pig were evaluated under electrical stimulation. Forskolin (1 microM) produced a massive release of ATP together with a positive inotropic response. Both 30 microM W-7 (N-(6-aminohexyl)-5-chloro-1 naphthalenesulfonamide.HCI), a calmodulin antagonist, and 30 microM vinblastine, a mitotic inhibitor, markedly inhibited the evoked release of ATP without affecting the evoked contraction. However, 100 microM N-ethylmaleimide abolished completely the basal and drug-evoked ATP release and further the evoked contraction. Both the ATP release and contraction evoked by ouabain (3 microM) were similarly affected by W-7, vinblastine and n-ethylmaleimide. The release of ATP, but not the contraction, evoked by forskolin was strongly suppressed by 10 microM okadaic acid, a protein phosphatase inhibitor. The suppression by okadaic acid of the evoked release was thoroughly antagonized in the presence of 0.01 microM PMA (phorbol 12-myristate 13-acetate), but not 10 microM H-7 (1-(5 isoquinolinesulfonyl)-2-methylpiperazine). These results suggest that forskolin, like ouabain, may dominantly cause the neuronal release of ATP from cardiac adrenergic nerves, although the possible participation of release from muscular sources cannot be ignored. PMID- 7498280 TI - Characteristics of caffeine-induced and spontaneous inward currents and related intracellular Ca2+ storage sites in guinea-pig tracheal smooth muscle cells. AB - Characteristics of caffeine-induced inward current (Icaf) and spontaneous transient inward current were examined in single smooth muscle cells isolated from guinea-pig trachea. When a pipette solution contained mainly CsCl, an application of 10 mM caffeine elicited transient Icaf at a holding potential of 60 mV. Spontaneous transient inward currents were recorded in about 15% of cells examined and were abolished by caffeine. Both were Cl- current activated by an increase in intracellular Ca2+ concentration (ICl-Ca). When 10 mM caffeine was puff-applied twice with various intervals, the amplitude of the second Icaf depended upon the period of the interval. The relationship between the amplitude and the interval represents the recovery time course of Icaf, which was significantly slowed by 30 microM cyclopiazonic acid. The Icaf was not significantly affected by addition of Cd2+. Removal of external Ca2+ did not affect the first Icaf but markedly reduced the second one, regardless of the presence of Cd2+. In conclusion, Icaf is evoked by activation of ICl-Ca via Ca2+ release. The recovery time course of Icaf indicates the refilling of Ca2+ storage sites by the cyclopiazonic acid-sensitive Ca2+ pump. The refilling at -60 mV depends strongly upon Ca2+ influx through the Cd(2+)-insensitive pathway. Spontaneous transient inward currents may be also due to ICl-Ca activated by spontaneous Ca2+ release from local storage sites. PMID- 7498281 TI - Substance P-(1-7) and substance P-(5-11) locally modulate dopamine release in rat striatum. AB - The effects of substance P, substance P-(1-7) and substance P-(5-11) on endogenous dopamine outflow in rat striatal slices were investigated. The dose response curves (0.01 nM to 10 microM) were bell-shaped, with significant increases at 0.1 and 1 nM but with no effect at higher concentrations. The tachykinin NK1 receptor agonist, [Sar9,Met(O2)11]substance P, significantly increased dopamine outflow at 10 and 100 nM. The effects of substance P or substance P-(5-11) and 25 mM KCl were additive. A negative interaction was observed with substance P-(1-7) and K+. The increase in dopamine outflow elicited by 1 nM substance P and substance P-(5-11) was reversed by the tachykinin NK1 receptor antagonist WIN 51,708 (17 beta-hydroxy-17 alpha-ethynyl-5 alpha androstano[3,2-b]pyrimido[1,2- alpha]benzimidazole) (25 and 250 nM), whereas only partial reversal was observed for the effect of substance P-(1-7). These results show that substance P fragments locally modulate striatal dopamine outflow and the mechanisms underlying this modulation may differ between N- and C-terminal fragments. PMID- 7498282 TI - Comparison of responses of canine pulmonary artery and vein to angiotensin II, bradykinin and vasopressin. AB - Responses to angiotensin II, bradykinin and arginine vasopressin were compared in helical strips of canine pulmonary arteries and veins. Angiotensin II contracted the artery but relaxed the vein strip. The artery contraction was augmented by indomethacin and aspirin and was abolished by losartan. The vein relaxation was not affected by endothelium denudation but was abolished by the cyclooxygenase inhibitors, a prostaglandin I2 synthase inhibitor and losartan. The bradykinin induced artery relaxation was inhibited by endothelium denudation, NG-nitro-L arginine (L-NA) or indomethacin and abolished by their combined treatment. The vein relaxation produced by bradykinin was endothelium-independent and was abolished by indomethacin. Vasopressin produced a slight relaxation in the arteries, which was abolished by endothelium denudation and L-NA. The vein relaxation produced by vasopressin was abolished by endothelium denudation and combined treatment with L-NA and indomethacin. It may be concluded that (1) activation of angiotensin AT1 receptor subtype in smooth muscle produces contraction and also relaxation due to prostaglandin I2 release; the former predominates over the latter in the artery, whereas only the latter is operative in the vein, (2) the bradykinin-induced relaxation is due to nitric oxide (NO) from the endothelium and prostaglandin I2 from subendothelial tissues in the artery and solely to prostaglandin I2 in the veins, and (3) the vasopressin induced relaxation is mediated by endothelial NO in the artery, and NO and prostaglandin I2 in the vein. PMID- 7498283 TI - Role of the VLA-4 integrin in leucocyte recruitment and bronchial hyperresponsiveness in the guinea-pig. AB - Guinea-pigs sensitized to ovalbumin develop airway eosinophilia and bronchial hyperresponsiveness to acetylcholine 24 h following ovalbumin challenge. We have used an antibody to the very late antigen-4 (VLA-4) integrin (HP 1/2) to investigate the role of this integrin in the recruitment of leucocytes to the airways and to study the link between eosinophilia and the development of bronchial hyperresponsiveness. In this model, HP 1/2 reduced the number of eosinophils recovered from bronchoalveolar lavage fluid, as well as reducing the activity of the enzyme eosinophil peroxidase in both bronchoalveolar lavage fluid and whole lung extracts. However, HP 1/2 did not reduce bronchial hyperresponsiveness. The results suggest that after antigen challenge, the VLA-4 integrin is involved in the recruitment of leucocytes to the airways at the level of the vascular endothelium, but increased numbers of eosinophils in lavage fluid or in the lung tissue are not an essential correlate of bronchial hyperresponsiveness in this model. PMID- 7498284 TI - Effect of delta 9-tetrahydrocannabinol on circling in rats induced by intranigral muscimol administration. AB - Unilateral injection of 25 ng of muscimol into the rat substantia nigra pars reticulata induced contralateral circling. Circling induced by 25 ng of muscimol was increased by simultaneous administration of 1 microgram of delta 9 tetrahydrocannabinol and abolished by 10 micrograms. Previous experiments have shown that delta 9-tetrahydrocannabinol enhances muscimol-induced catalepsy in the globus pallidus. These results identify another brain area in which synergism between delta 9-tetrahydrocannabinol and muscimol can occur and show that delta 9 tetrahydrocannabinol can enhance an excitatory as well as inhibitory GABAergic motor effect. The data support the hypothesis that cannabinoids can facilitate transmission along central gamma-aminobutyric acid (GABA) releasing pathways. PMID- 7498285 TI - S-adenosyl-L-homocysteine and 5'-methylthioadenosine inhibit binding of [3H]flunitrazepam to rat brain membranes. AB - The effect of inhibitors of transmethylation reactions, S-adenosylhomocysteine and 5'-methylthioadenosine, on [3H]flunitrazepam-specific binding to the rat brain membranes has been investigated. Both S-adenosylhomocysteine and 5' methylthioadenosine are able to inhibit binding with Ki values of 7.9 microM and 15.8 microM respectively. These compounds therefore may be candidate endogenous benzodiazepine-receptor ligands. In light of these and other data possible correlations between phospholipid methylation and gamma-aminobutyric acid (GABA) receptor function are discussed. PMID- 7498286 TI - The effect of 2-amino-3-phosphonopropionic acid (AP-3) in the gerbil model of cerebral ischaemia. AB - The effect of 2-amino-3-phosphonopropionate (AP-3), a metabotropic glutamate receptor antagonist on behavioral and histological changes following global ischaemia was investigated on the Mongolian gerbil. Ischaemia was induced by bilateral carotid occlusion for 5 min. AP-3 was administered i.p. (25 or 250 mg/kg) 30 min before and 24 h after surgery. Significant neuroprotection was observed 96 h after surgery to cells in the CA1 region of the hippocampus in drug treated animals. AP-3 (250 mg/kg) significantly attenuated the increase in locomotor activity measured 72 h after surgery. These results suggest that metabotropic glutamate receptors play a role in the neurodegeneration seen following ischaemia. PMID- 7498287 TI - Involvement of peripheral presynaptic inhibition in the reduction of sympathetic tone by moxonidine, rilmenidine and UK 14304. AB - We studied the possibility that presynaptic inhibition of transmitter release from postganglionic sympathetic neurons contributes to the overall reduction of sympathetic tone produced by moxonidine, rilmenidine and 5-bromo-6-(2-imidazolin 2-ylamino)-quinoxaline tartrate (UK 14304). In pithed rabbits without electric stimulation, moxonidine, rilmenidine and UK 14304 caused a long-lasting, > 10 min, increase in arterial pressure. Heart rate was not changed. In pithed rabbits in which sympathetic tone was created by electric stimulation through the pithing rod (2 Hz), the same doses of moxonidine, rilmenidine and UK 14304 caused only a brief, < 10 min, blood pressure rise. Heart rate was decreased, as were the plasma concentrations of noradrenaline and adrenaline. Dose-response curves for the effects on the plasma noradrenaline concentration (stimulated pithed rabbits) were compared with previously obtained dose-response curves for depression of renal sympathetic nerve activity (conscious rabbits). For each drug, the curve describing peripheral presynaptic inhibition and the curve describing central sympathoinhibition were very similar. Both the power and the dose dependence of the peripheral inhibitory effect support its contribution to the overall decrease in sympathetic tone produced by clonidine-like drugs in intact animals. The peripheral effect in all likelihood consists in activation of presynaptic alpha 2 autoreceptors. The agreement of the dose-response curves for the peripheral and for the central effect supports the view that the central effect, like the peripheral one, is mediated through alpha 2-adrenoceptors. PMID- 7498288 TI - Modulation of rat neocortical high-voltage spindle activity by 5-HT1/5-HT2 receptor subtype specific drugs. AB - To investigate the role of serotonin (5-hydroxytryptamine; 5-HT) receptors in the modulation of rat thalamocortical oscillations, we studied the effects of 5-HT1/5 HT2 receptor subtype specific drugs on neocortical high-voltage spindle activity in adult male rats. A 5-HT1A receptor agonist, 8-hydroxy-2-(di-n propylamino)tetralin (8-OH-DPAT) (0.03, 0.1, 0.3 and 1.0 mg/kg s.c.), had no effect on neocortical high-voltage spindle activity. Furthermore, a mixed 5-HT1/5 HT2 receptor antagonist, methysergide (1.0, 5.0 and 15.0 mg/kg i.p.), had no effect, whereas a non-specific mixed 5-HT1/5-HT2 receptor antagonist, methiothepin (0.2, 1.0 and 5.0 mg/kg i.p.), significantly increased neocortical high-voltage spindles. Of the 5-HT2 receptor antagonists, ritanserin (0.1, 1.0 and 5.0 mg/kg s.c.) had no effect, whereas ketanserin (1.0, 5.0 and 20.0 mg/kg s.c.) increased neocortical high-voltage spindles, but only at the highest dose used. A 5-HT2 receptor agonist, 1-(2,5-dimethoxy-4-iodophenyl)-2-aminopropane (DOI) (0.5, 1.0 and 2.0 mg/kg s.c.), at the two highest doses significantly decreased neocortical high-voltage spindle activity, and this effect was blocked by the 5-HT2 receptor antagonists, ketanserin (1.0, 5.0 and 20.0 mg/kg s.c.) and ritanserin (1.0 and 5.0 mg/kg s.c.), as well as by methiothepin (0.2, 1.0 and 5.0 mg/kg i.p.) and methysergide (1.0, 5.0 and 15.0 mg/kg i.p.). Furthermore, unilateral intrathalamic infusions, but not intrahippocampal control infusions, of DOI (10 and 50 micrograms/1.0 microliters/rat) decreased neocortical high voltage spindle activity and systemic administration of ketanserin (20.0 mg/kg s.c.) completely blocked this effect. The present results suggest that (1) the serotonergic system modulates rat thalamocortical oscillations as measured by neocortical high-voltage spindle activity, (2) activation of 5-HT2 receptors, possibly located in the thalamus, with a specific 5-HT2 receptor agonist, DOI, causes a reduction in rat neocortical high-voltage spindle activity. PMID- 7498289 TI - Characterisation of functional endothelin receptors in the canine isolated perfused spleen. AB - The endothelin receptor subtypes involved in the vasoconstriction, capsular smooth muscle contraction, prostaglandin E2 and prostacyclin release induced by endothelin-1 have been investigated in the canine isolated perfused spleen using both the endothelin ETA receptor antagonist FR 139317 and the endothelin ETB receptor agonist IRL 1620. THe isolated canine spleen was perfused with warmed (37 degrees C) and oxygenated (95% O2/5% CO2) Krebs solution at constant flow with continuous recording of splenic arterial perfusion pressure and spleen weight. Samples of splenic venous effluent were collected to determine the amounts of prostaglandin E2 and prostacyclin, measured by radioimmunoassay. Endothelin-1 (4-200 pmol) and IRL 1620 (20-1000 pmol) dose-dependently increased splenic arterial perfusion pressure but the former was more potent on a molar basis (the molar dose ratio IRL/endothelin-1 required to increase splenic arterial vascular resistance by 25% was approximately 33). The infusion of the nitric oxide inhibitor N omega-nitro-L-arginine methyl ester (10 microM), but not of the enantiomer N omega-nitro-D-arginine methyl ester, significantly potentiated the increase in splenic arterial vascular resistance induced by endothelin-1. The infusion of FR 139317 (1 microM) markedly attenuated the increased splenic arterial perfusion pressure induced by endothelin-1 without affecting that evoked by IRL 1620. At the highest dose (200 pmol), endothelin-1 induced a small but significant capsule contraction as reflected by the reduction in the spleen weight. The infusion of FR 139317 (1 microM) abolished this contractile effect. IRL 1620 (in doses up to 1000 pmol) did not significantly affect the capsule tone.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7498290 TI - Long-term antidepressant treatments alter 5-HT2A and 5-HT2C receptor-mediated hyperthermia in Fawn-Hooded rats. AB - We have recently demonstrated that hyperthermia induced by 1-(2,5-dimethoxy-4 iodophenyl)-2-aminopropane (DOI) and m-chlorophenylpiperazine (m-CPP) are separately mediated by selective stimulation of 5-HT2A and 5-HT2C receptors, respectively in Wistar rats. Furthermore, hyperthermia induced by either DOI or m CPP was found to be significantly less in Fawn-Hooded rats (a rat strain suggested to represent a genetic model of depression) relative to Wistar rats. In the present study, we studied the effects of long-term antidepressant treatments on DOI (2.5 mg/kg)-induced and m-CPP (2.5 mg/kg)-induced hyperthermia in male Fawn-Hooded rats. Long-term (21 days) treatment with the tricyclic antidepressants, imipramine or clomipramine (each 5 mg/kg/day), attenuated DOI induced hyperthermia, while m-CPP-induced hyperthermia was accentuated. On the other hand, long-term (21 days) treatment with the monoamine oxidase type-A inhibiting antidepressant, clorgyline (1 mg/kg/day), did not modify m-CPP-induced hyperthermia, but significantly attenuated DOI-induced hyperthermia. These findings demonstrate that long-term antidepressant treatments alter 5-HT2A and 5 HT2C receptor-mediated hyperthermia in a genetic animal model of depression. PMID- 7498291 TI - Suppression of oro-facial movements by rolipram, a cAMP phosphodiesterase inhibitor, in rats chronically treated with haloperidol. AB - We investigated the effects of rolipram, a selective cyclic adenosine 3',5' monophosphate phosphodiesterase type IV inhibitor, and isobutylmethylxanthine, a nonselective phosphodiesterase inhibitor, on purposeless spontaneous chewing movements and tongue protrusions produced by 24 weeks treatment with haloperidol decanoate (25 mg/kg every 4 weeks i.m.) in rats, to examine our hypothesis that restoration of striatal cyclic adenosine 3',5'-monophosphate levels previously reduced due to dopamine D2 receptor supersensitivity, may suppress these movements. Tests were performed 8 weeks after the final injection. Haloperidol treatment significantly increased dyskinetic movements and striatal dopamine D2 receptor density compared with controls. Rolipram (0.1-1.0 mg/kg i.p.) suppressed these movements in a dose-dependent manner, whereas isobutylmethylxanthine (2 mg/kg i.p.) only slightly suppressed the syndrome and doses higher than 5 mg/kg i.p. produced other intensive movements. These results support our hypothesis and suggest that rolipram may have a therapeutic effect on tardive dyskinesia. PMID- 7498292 TI - Improved glucose metabolism following blockade of angiotensin converting enzyme but not angiotensin AT1 receptors. AB - This study compared the effect of benazepril, an angiotensin converting enzyme inhibitor ([S-(R*,R*)]-3-[[1-(ethoxycarbonyl)-3-phenylpropyl]amino]-2,3,4,5- tetrahydro-2-oxo) to valsartan, an angiotensin AT1 receptor antagonist ((S)-N valeryl-N-[2'-(1H-tetrazol-5-yl)biphenyl-4-yl]-methyl-valin e) on glucose metabolism and plasma lipid levels in 11- to 12-week-old conscious spontaneously hypertensive rats. Intraperitoneal infusion of benazepril or valsartan at 1, 3 and 10 mg/kg/day produced dose-related reductions in systolic blood pressure after 12 weeks which were not significantly different from each other. During the infusion, valsartan-treated animals gained weight at the same rate as controls, but all infusion rates of benazepril significantly suppressed normal weight gain, despite both compounds having similar antihypertensive effects. At the end of the 12-week treatment period, benazepril (3 and 10 mg/kg/day) significantly increased glucose disposal but did not significantly affect insulin disposal in insulin/glucose tolerance tests. In contrast, none of the infusion rates of valsartan significantly affected glucose or insulin disposal. Finally, compared to controls benazepril and valsartan were without effect on the fasting basal plasma concentrations of glucose, insulin, triglycerides, total cholesterol and K+. The results demonstrate that angiotensin converting enzyme inhibition and angiotensin AT1 receptor antagonism have similar antihypertensive effects, but express differences on body weight gain and insulin-stimulated glucose disposal in the conscious spontaneously hypertensive rat. PMID- 7498293 TI - Evidence for alterations in presynaptic serotonergic function during withdrawal from chronic cocaine in rats. AB - The effects of repeated cocaine administration on serotonin (5-hydroxytryptamine, 5-HT) function were investigated by comparing the corticosterone response to 5-HT receptor agonists in cocaine-treated and vehicle-treated rats. Male rats were fitted with indwelling jugular catheters and received cocaine (15 mg/kg i.p., b.i.d.) or saline for 7 days. Rats were challenged with either saline, the 5-HT releaser fenfluramine (1.2 mg/kg i.v.), the 5-HT1A receptor agonist 8-hydroxy-2 (di-n-propylamino)tetralin (8-OH-DPAT; 50 micrograms/kg i.v.), or the 5-HT2A/2C receptor agonist 1-(2,5-dimethoxy-4-iodophenyl)-2-aminopropane (DOI; 100 micrograms/kg i.v.) 42 h and 8 days after the final chronic treatment. Repeated blood samples were withdrawn immediately before and at 15, 30 and 60 min after acute challenge injections. All 5-HT receptor agonists increased plasma corticosterone, but the fenfluramine-induced rise in corticosterone was significantly attenuated in cocaine-treated rats withdrawn for 42 h. This blunted response to fenfluramine exhibited only partial recovery when examined at 8 days postchronic treatment. Corticosterone responses to 8-OH-DPAT and DOI were not affected by cocaine exposure. Our data suggest that chronic cocaine produces deficits in presynaptic 5-HT function, and alterations in 5-HT neurotransmission may underlie the dysphoria experienced by abstinent cocaine users. Neuroendocrine challenge tests should be performed in human addicts to evaluate potential 5-HT dysfunction associated with cocaine abuse. PMID- 7498294 TI - Characterization of contractile adrenoceptors in the human umbilical artery. AB - Adrenoceptors mediating contraction in ring segments of human umbilical arteries from normal term pregnancies were investigated in vitro. Contraction was elicited by (order of potency indicated): noradrenaline = the alpha 2-adrenoceptor agonist oxymetazoline >> the alpha 1-adrenoceptor agonist phenylephrine. The alpha 1 adrenoceptor antagonist prazosin antagonized the contraction elicited by noradrenaline and phenylephrine. The alpha 2-adrenoceptor antagonist rauwolscine antagonized the contraction elicited by noradrenaline and oxymetazoline. Oxymetazoline had an efficacy 5 times higher than that of noradrenaline and the 5 hydroxytryptamine receptor antagonist methysergide antagonized the contraction elicited by oxymetazoline. It is suggested that the contractile adrenoceptors in the human umbilical artery consist of both alpha 1 and alpha 2 subtypes. Furthermore, the contractile effect of oxymetazoline seems to be mediated via both alpha 2-adrenoceptors and 5-hydroxytryptamine receptors. PMID- 7498295 TI - First delineation of 5-HT1A receptors in human brain with PET and [11C]WAY 100635. AB - The selective 5-HT1A receptor radioligand, [11C]WAY-100635 ([11C]N-2-(4-(2 methoxyphenyl)-1-piperazinyl)ethyl)-N-2- pyridyl)cyclohexanecarboxamide), has been injected intravenously into healthy male volunteers and studied by PET (positron emission tomography). The results provide the first delineation of 5 HT1A receptors in living human brain and demonstrate the potential to use [11C]WAY-100635 for the study of central 5-HT1A receptors in patients with psychiatric and neurological disorders and for the investigation of the pharmacology of drugs acting on the central nervous system. PMID- 7498296 TI - Fenfluramine's appetite suppression and serotonin neurotoxicity are separable. AB - To determine whether fenfluramine's anorectic and neurotoxic effects could be dissociated, rats were treated with fenfluramine or the serotonin transporter blocker fluoxetine, alone or in combination. Fenfluramine alone produced anorexia, weight loss and lasting depletions of brain serotonin axon markers. Fluoxetine prevented fenfluramine-induced long-term serotonergic deficits, yet did not diminish fenfluramine's acute anorectic effects. These findings indicate that fenfluramine's anorectic and neurotoxic actions are distinct and separable. PMID- 7498297 TI - Cromakalim- and RP 49356-induced 42K+ and 86Rb+ efflux in rat myometrium. AB - Previous studies had failed to observe cromakalim-induced 42K+ or 86Rb+ efflux from the myometrium of the pregnant rat in contrast to positive findings in other smooth muscles. In the current study, in myometrium from the non-pregnant rat, cromakalim (10 microM) and RP 49356 ([(+/-)-N-methyl-2-(3-pyridyl) tetrahydrothiopyran-2-carbothioamid e-1-oxide); 10 microM) induced small increases in 42K+ or 86Rb+ efflux but much less than did oxytocin (20 nM) or KCl (20 mM). The cromakalim-induced increase in 42K+ efflux was enhanced 3.5-fold in the presence of KCl (20 mM) plus (+)-cis-diltiazem (3 microM), a property shared by RP 49356. Glibenclamide (10 microM) partially reduced the cromakalim-induced 42K+ efflux, in the presence of KCl and (+)-cis-diltiazem, but did not affect the KCl-induced 42K+ efflux. The data provides further support for the idea that cromakalim and RP 49356 are able to open potassium channels in rat myometrium. It would appear that their actions in this tissue are dependent on the extracellular K+ concentration and/or membrane potential. PMID- 7498299 TI - Inhibitory effect of YM060 on 5-HT3 receptor-mediated depolarization in colonic myenteric neurons of the guinea pig. AB - We used conventional intracellular recording methods to examine the effects of YM060 [(-)-(R)-5-[(1-methyl-1H-indol-3-yl)carbonyl]-4,5,6,7- tetrahydro-1H benzimidazole monohydrochloride), a novel 5-HT3 receptor antagonist, on 5 hydroxytryptamine (5-HT, serotonin)-evoked fast membrane depolarization in myenteric neurons of the guinea pig distal colon, and compared its potency to that of other 5-HT3 receptor antagonists. Microapplication of 5-HT from fine tipped pipettes evoked both fast and slowly activating depolarizing responses in 78% and 40% of colonic myenteric neurons, respectively. The selective 5-HT3 receptor agonist 2-methyl-5-HT applied with short pressure pulses (100-300 ms) mimicked the fast but not the slow response. The 5-HT3 receptor antagonists YM060, granisetron and ondansetron suppressed the 5-HT-evoked fast response in 98% of colonic myenteric neurons in a concentration-dependent manner with pIC50 values of 8.62, 7.77 and 6.90, respectively. Methysergide and GR113808 did not affect the fast responses at concentrations sufficient to block 5-HT1, 5-HT2 and 5-HT4 receptors, respectively. YM060 did not affect the slowly activating response to 5-HT or any other electrophysiological parameter of the neurons including resting membrane potential, input resistance and the amplitude of action potentials evoked by injection of depolarizing current. Stimulus-evoked fast excitatory postsynaptic potentials were unchanged by YM060 at concentrations up to 10(-8) M, excluding any possible local anesthetic or anticholinergic effects of YM060. The results confirm that the fast component of the two depolarizing responses to 5-HT in colonic myenteric neurons is mediated by 5-HT3 receptors.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7498298 TI - Effect of diterpenoid alkaloids on cardiac sympathetic efferent and vagal afferent nerve activity. AB - The diterpenoid alkaloid, lappaconitine, at a dose of 150 micrograms/kg (i.v.) increased cardiac vagal afferent nerve activity (16.2%) and reduced cardiac sympathetic efferent nerve activity (12.5%). A polar analog, N deacetyllappaconitine, at this same dose, increased cardiac vagal afferent nerve activity (40%) and reduced cardiac sympathetic efferent nerve activity (23.5%). Both of these agents also reduced arterial blood pressure and heart rate. A larger dose of lappaconitine (300 micrograms/kg i.v.) produced the same changes in nerve activities and cardiac function as the lower dose. Two other structurally related agents, lycoctonine and aconine, failed to alter these variables in doses up to 300 micrograms/kg. These data suggest that certain diterpenoid alkaloids activate autonomic reflex receptors, including cardiac reflex receptors. The polar agent, N-deacetyllappaconitine, appears to be more effective on cardiac reflex receptors than the non-polar agent, lappaconitine. Such agents may be useful in the treatment of hypertension. PMID- 7498301 TI - Effects of dopamine D1 and D2 receptor antagonists on cocaine-induced place preference conditioning in preweanling rats. AB - The effects of dopamine D1 and D2 receptor antagonists on the reward processes of 10- and 17-day-old rats were assessed using the conditioned place preference paradigm. Conditioning and testing were conducted in a three-compartment chamber, with each end compartment having its own distinct tactile and odor cues (almond and lemon). During six experiments, 10- and 17-day-old rats (age at initial conditioning) were injected intraperitoneally with either saline, the dopamine D1 receptor antagonist R(+/-)-SCH 23390 hydrochloride (0.01-1.0 mg/kg), or the dopamine D2 receptor antagonists (+/-)-sulpiride (1-100 mg/kg) or S(-) eticlopride hydrochloride (0.1-0.5 mg/kg) 30 min prior to being injected with cocaine hydrochloride (20 mg/kg) or saline. After the latter injections, rats were immediately confined in the lemon-scented (nonpreferred) compartment for 30 min. On the alternate conditioning day, rats were injected with saline and confined in the almond-scented compartment. On the third day (i.e., the test day), rats were given saline and allowed free access to the entire chamber for 15 min. The results showed that the dopamine D1 receptor antagonist SCH 23390 blocked the cocaine-induced place preference conditioning of both 10- and 17-day old rats. Surprisingly, the dopamine D2 receptor antagonists sulpiride and eticlopride blocked the place preference conditioning of 10-day-old rats, while leaving the 17-day-old rats unaffected. These results indicate that dopamine D1 receptors are critically involved in the reward processes of preweanling rats, but that the importance of dopamine D2 receptors changes across ontogeny. PMID- 7498300 TI - Effects of dopamine on snail neurones. AB - The pharmacological features of dopamine receptors in identifiable giant neurone types of a snail (Achatina fulica Ferussac) were studied. Under voltage clamp, two neurone types, LVMN (left ventral multiple spike neurone) and d-RPeAN (dorsal right pedal anterior neurone), produced an inward current (Iin) in response to dopamine, (-)-noradrenaline and epinine, whereas v-LCDN (ventral-left cerebral distinct neurone) produced an outward current (Iout) in response to dopamine and epinine. Mammalian dopamine receptor agonists, fenoldopam (dopamine D1-like receptor agonist), (+/-)-SKF 38393 (1-phenyl-2,3,4,5-tetrahydro-1H-3-benzazepine 7,8- diol) (D1-like), apomorphine (D2-like), (-)-quinpirole (D3 and D4) and methylergometrine showed slight or no effect. (+/-)-SKF 83566 ((+/-)-7-bromo-8 hydroxy-3-methyl-1-phenyl-2,3,4,5-tetrahydro-1H-3-benza zepine) (dopamine D1-like receptor antagonist) and (+)-UH 232 (cis-(+)-5-methoxy-1-methyl-2-(di-n propylamino)tetralin) (D3 and D2) non-competitively inhibited the Iin of LVMN and d-RPeAN, but (+/-)-sulpiride (D2-like) was without effect. In contrast, (+/-) sulpiride competitively inhibited Iout of v-LCDN, (+)-UH 232 non-competitively inhibited Iout of v-LCDN but (+/-)-SKF 83566 was without effect. H-7 (1-(5 isoquinolinesulfonyl)-2-methylpiperazine) (non-selective protein kinase inhibitor) inhibited Iin of LVMN and d-RPeAN, but did not affect Iout of v-LCDN. Dopamine-induced Iin was Na(+)-dependent; Iout was K(+)-dependent. Ouabain did not affect these currents. We propose that the pharmacological features of Achatina neuronal dopamine receptors are not fully comparable to those of mammals, although intracellular signal transduction systems linked with dopamine receptors may similarly exist in different animal species. PMID- 7498302 TI - (S)-WAY 100135, a 5-HT1A receptor antagonist, prevents the impairment of spatial learning caused by intrahippocampal scopolamine. AB - Scopolamine, 3.75 micrograms/microliters infused bilaterally into the CA1 region of the dorsal hippocampus 10 min before each training session, impaired choice accuracy but had no effect on choice latency or errors of omission in rats trained in a two-platform spatial discrimination task. Administered subcutaneously at 3 and 10 mg/kg 30 min before each training session, N-tert butyl-3-4-(2-methoxyphenyl)piperazin-1-yl-2-phenylpropanami de dihydrochloride ((S)-WAY 100135), a 5-HT1A receptor antagonist, prevented the impairment of choice accuracy induced by intrahippocampal scopolamine. No subcutaneous dose of (S)-WAY 100135 by itself modified the acquisition of spatial learning. Administered into the dorsal hippocampus 15 min before each training session, (S) WAY 100135 at doses of 0.2, 1 and 5 micrograms/microliters did not affect the acquisition of spatial learning but dose dependently prevented the impairment of choice accuracy caused by scopolamine, 3.75 micrograms/microliters infused into the same area. These findings suggest that blockade of 5-HT1A receptors can compensate the loss of cholinergic excitatory input on pyramidal cells, probably by favouring the action of other excitatory transmitters. PMID- 7498303 TI - Mechanisms mediating NG-nitro-L-arginine methyl ester-induced hypophagia in mice. AB - NG-Nitro-L-arginine methyl ester (50 mg/kg s.c.), an inhibitor of nitric oxide (NO) synthase, has been reported to increase brain serotonin (5 hhydroxytryptamine, 5-HT) metabolism and induce hypophagia. Conversely, enhanced NO synthase activity is found to be accompanied by a decrease in 5-HT level. This negative correlation between NO and 5-HT in the regulation of food intake was further studied in mice. 5-HT depletion by p-chlorophenylalanine (250 mg/kg i.p., twice daily for 2 days) failed to antagonize the hypophagic effect of NG-nitro-L arginine methyl ester. Similarly, treatment with the NO synthesis precursor, L arginine (1000 mg/kg s.c.), did not reverse the anorexia induced by fenfluramine (10 mg/kg s.c.), a 5-HT releaser/uptake inhibitor. Pretreatment with (-) pindolol, methylsergide and ritanserin had no effect on the hypophagic action of NG-nitro-L-arginine methyl ester, suggesting the lack of involvement of 5-HT1 and 5-HT2 receptors. The selective neuronal NO synthase inhibitor, 7-nitroindazole (12.5-50.0 mg/kg i.p.), however, did not exhibit any hypophagic effect whilst NG nitro-L-arginine methyl ester increased gastric retention, which may subsequently induce satiety. Moreover, the hypophagic effect of NG-nitro-L-arginine methyl ester, which was unassociated with changes in water intake and malaise induction, was also unattenuated by cholecystokinin (CCK) receptor antagonists, devazepide (10 mg/kg i.p.) and PD 135,158 ([1S-[1 alpha,2 beta[S*(S*)],4 alpha ]]-4-[[2-[[3 (1 H-indol-3-yl)-2-methyl-1-oxo-2-[[[(1,7,7- trimethylbicyclo[2.2.1]hept-2 yl)oxy]carbonyl]amino]propyl]amino] -1-phenylethyl] amino]-4-oxo-butanoic acid N methyl-D-glucamine salt; 1 mg/kg i.p.).(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7498304 TI - GABAA receptor antagonism in the extended amygdala decreases ethanol self administration in rats. AB - The present experiments examined the role of the extended amygdala GABAA receptors in the regulation of ethanol consumption in rats. The areas of the extended amygdala studied included the central nucleus of the amygdala, the bed nucleus of the stria terminalis, and the shell of the nucleus accumbens. The effects of bilateral microinjections of a competitive GABAA receptor antagonist, 2-(3-carboxypropyl)-3-amino-6-(4-methoxyphenyl)pyridazinium bromide (SR 95531), on ethanol consumption were assessed in Wistar rats that were trained to respond for oral ethanol (10% w/v) in a two-lever, free-choice operant task during 30-min sessions using a saccharin fading procedure. Injections of SR 95531 into the central amygdaloid nucleus decreased ethanol responding significantly at doses of 2 and 4 ng without affecting water responding. SR 95531 injections into the bed nucleus of the stria terminalis reduced ethanol responding significantly at the 8 ng and 16 ng dose, while only the 16 ng dose produced a significant effect in the shell of the nucleus accumbens. Cumulative response patterns showed that intra amygdaloid injections did not disrupt the initiation of responding. Injections into the bed nucleus of the stria terminalis and the nucleus accumbens, however, suppressed both ethanol and water responding at the highest SR 95531 doses during the first minutes. These findings suggest that GABAA receptors in the extended amygdala may be involved in the mediation of some aspects of ethanol reward. PMID- 7498305 TI - Beneficial effects of the endothelin receptor antagonist bosentan on myocardial and endothelial injury following ischaemia/reperfusion in the rat. AB - The effects of bosentan, a nonpeptide endothelin receptor antagonist, on endothelin-induced changes in coronary flow and myocardial ischaemic and reperfusion injury were investigated in the Langendorff perfused rat isolated heart. Endothelin-1 (0.012-0.4 nmol) evoked dose-dependent reduction in coronary flow, which was attenuated by bosentan (1.0-10 microM) in a concentration-related fashion. The inhibitory effect of bosentan lasted more than 30 min. The endothelin ETB receptor agonist Suc-[Glu9,Ala11,15]endothelin-1-(8-21) (IRL 1620) increased coronary flow in the absence but not in the presence of bosentan. In hearts subjected to 30 min of global ischaemia followed by 30 min of reperfusion, the recoveries of the left ventricular developed pressure, dP/dtmax, and coronary flow were significantly larger in a group given bosentan 10 microM at the start of ischaemia (92 +/- 7%, 98 +/- 8% and 83 +/- 5%, respectively) than in a vehicle treated group (70 +/- 4%, 70 +/- 6% and 42 +/- 2%, respectively) at the end of the reperfusion period. During the reperfusion period, left ventricular end diastolic pressure was significantly lower in the bosentan group than in the vehicle group. The area of no-reflow in the bosentan group was 7 +/- 3% of left ventricle compared to 21 +/- 2% in the vehicle group (P < 0.01). Acetylcholine induced endothelium-dependent vasodilatation was significantly reduced after ischaemia and reperfusion in the vehicle group but not in the bosentan group. It is concluded that bosentan attenuates the coronary vasoconstrictor effect elicited by endothelin and reduces ischaemia/reperfusion-induced myocardial and endothelial injury in the rat isolated heart.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7498306 TI - Intermittent and chronic morphine treatment induces long-lasting changes in delta opioid receptor-regulated acetylcholine release in rat striatum and nucleus accumbens. AB - Intermittent treatment of rats with morphine (10 mg/kg s.c., once daily) caused an increase (of about 30%) of the electrically evoked release of [14C]acetylcholine from cholinergic interneurons of superfused striatal slices 1 21 days after morphine withdrawal. Similarly, chronic treatment with escalating doses of morphine (5-50 mg/kg s.c., 3 times daily), causing physical dependence (unlike intermittent treatment), resulted in an enduring enhanced response of these neurons towards depolarization. Following chronic morphine treatment this adaptive increase of acetylcholine release was associated with a slight but long lasting decrease of the (delta-opioid receptor-mediated) maximal inhibitory effect of [Met5]enkephalin, whereas upon intermittent drug treatment delta-opioid receptor desensitization was observed 1 day after opiate withdrawal only. Also in slices of the nucleus accumbens both intermittent as well as chronic morphine administration caused a long-lasting increase of the electrically evoked [14C]acetylcholine release. Therefore, we hypothesize that an enhanced (re)activity of striatal and accumbal cholinergic neurons, which are regulated by dopaminergic neurons of the ventral mesencephalon, may represent a long-lasting neuroadaptive effect of morphine (and possibly other drugs of abuse) playing a crucial role in behavioral sensitization associated with enhanced vulnerability to drugs of abuse. PMID- 7498307 TI - Ethanol withdrawal is associated with increased extracellular glutamate in the rat striatum. AB - Extracellular glutamate was measured by microdialysis in the striatum of ethanol dependent, freely behaving rats following withdrawal from chronic ethanol treatment. Within 12 h from withdrawal, extracellular glutamate rose to 255% of that in control, chronic sucrose-treated rats. Glutamate output remained elevated for the subsequent 12 h and returned to control levels within 36 h from the interruption of the treatment. The changes in glutamate were time-locked to the overt physical signs of withdrawal. In 12-h ethanol-withdrawn rats an ethanol challenge suppressed the withdrawal signs and reduced the extracellular glutamate. The NMDA receptor antagonist, dizocilpine, reduced both the physical signs of withdrawal and glutamate output. In contrast, diazepam reduced the withdrawal signs but failed to change the glutamate levels. These findings suggest that the increased extraneuronal glutamate reflects overactivity of excitatory neurotransmission during withdrawal. Furthermore, they provide a biochemical rationale for the use of NMDA receptor antagonists and ethanol itself in the treatment of ethanol withdrawal syndrome. PMID- 7498309 TI - Neuroprotective and antioxidant activities of HU-211, a novel NMDA receptor antagonist. AB - This study examines the ability of (+)-(3S,4S)-7-hydroxy-delta 6 tetrahydrocannabinol-1,1-dimethylheptyl (HU-211), a non-competitive NMDA receptor antagonist to: (1) rescue neurons in culture from injury evoked by sodium nitroprusside, hydrogen peroxide (H2O2) and oxygen glucose deprivation; and (2) scavenge reactive oxygen species in vitro. Qualitative and quantitative assessments of cell survival have indicated that: (1) Neuronal cell injury produced following deprivation of oxygen and glucose was significantly attenuated by 5 microM HU-211. (2) Glial and neuronal cell damage induced by sodium nitroprusside was markedly ameliorated by 10 microM HU-211. (3) HU-211 reduced protein oxidation initiated by gamma irradiation, and scavenged peroxyl radicals. (4) HU-211 carries an oxidation potential of 550 mV. These findings suggest that HU-211 holds a unique position among putative neuroprotectant agents in that it combines NMDA receptor antagonistic activity and free radical scavenging abilities in a single molecule. PMID- 7498310 TI - Topical glucocorticoids inhibit neurogenic inflammation: involvement of lipocortin 1. AB - Topical glucocorticoid treatment (betamethasone-17-valerate (0.018 mg/cm2, 3 h pretreatment) significantly inhibited neurogenic oedema formation induced by electrical antidromic stimulation (2 Hz, 15 V, 0.1 ms for 5 min) of the rat saphenous nerve; a response mediated by neuropeptides released from activated capsaicin-sensitive sensory C-fibres. Oedema formation was estimated by measurement of extravasation of i.v. injected 125I-albumin into skin. The inhibitory effect of the topical glucocorticoid was reversed by passive immunisation of rats with polyclonal antibody to the glucocorticoid-inducible anti-inflammatory protein lipocortin 1 (1 ml/kg, s.c., 24 h pretreatment) whilst a non-immune serum was without effect. Similarly the glucocorticoid receptor antagonist RU38486 (20 mg/kg, 2 and 20 h pretreatment) abrogated the response indicating specific binding to glucocorticoid receptors. Topical glucocorticoid treatment also inhibited the oedema produced by intradermal substance P (0.1 nmol) in the dorsal skin of rats. Topical glucocorticoid inhibited neurogenic oedema formation partly through a mechanism dependent upon lipocortin 1. This inhibition may be partly due to a post-junctional effect upon substance P activity/binding however a pre-junctional component cannot be excluded. PMID- 7498308 TI - Chronic NMDA receptor stimulation: therapeutic implications of its effect on adenosine A1 receptors. AB - It is known that stimulation of adenosine A1 receptors has a modulatory effect on the excitability of postsynaptic NMDA receptors. Conversely, acute stimulation of NMDA receptors results in release of adenosine via calcium-independent mechanisms. These findings indicate a close functional relationship between these receptors. It is, therefore, possible that chronic, low level stimulation of the NMDA receptor may have a negative impact on these modulatory processes. To investigate this possibility, we have subjected C57BL mice either to an acute injection of a N6-cyclopentyladenosine (CPA, 0.01 mg/kg) or deoxycoformycin (1 mg/kg) followed by a convulsant dose of N-methyl-D-aspartate (NMDA) (60 mg/kg) or to chronic, low level (20 mg/kg i.p. daily) exposure to NMDA for 8 weeks. One day after the last injection of NMDA, animals were injected either with a convulsant dose of NMDA alone, or with either CPA at 0.001 or 0.01 mg/kg, or with 1 mg/kg deoxycoformycin followed 15 min later by 60 mg/kg NMDA. Neither CPA nor deoxycoformycin were protective when NMDA was given acutely at 60 mg/kg. Chronic treatment with NMDA alone or chronic administration of NMDA followed by 0.001 mg/kg CPA had no significant effect on mortality following a convulsant dose of NMDA. However, when the chronic regimen of NMDA was followed by either 0.01 mg/kg CPA or 1 mg/kg deoxycoformycin, mortality was reduced to 10% (CPA), or eliminated completely (deoxycoformycin). Moreover, combination of chronic NMDA treatment with either CPA (both doses) or deoxycoformycin produced a significant improvement in other measures, i.e., seizure onset, intensity of neurological impairment, and extension of time to death.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7498311 TI - Differences in the effects of ketanserin and GR127935 on 5-HT-receptor mediated responses in rabbit saphenous vein and guinea-pig jugular vein. AB - Ketanserin has higher affinity for 5-HT1D alpha receptors compared to 5-HT1D beta receptors, whereas, GR127935 (N-[4-methoxy-3-(4-methyl-1-piperazinyl)phenyl] 2(methyl-4(-(5-methyl- 1,2,4-oxadiazol-3-yl)[1,1-biphenyl]-4-carboxamide), a novel and selective 5-HT1D receptor antagonist, has higher affinity for 5-HT1D beta receptors compared to 5-HT1D alpha receptors. In the present study, we compared the effects of ketanserin and GR127935 on sumatriptan-induced responses of rabbit saphenous vein and guinea-pig jugular vein. In rabbit saphenous vein, contractile responses elicited by sumatriptan were antagonised by ketanserin (pA2 = 6.76) and GR127935 (apparent pA2 = 9.4). In guinea-pig jugular vein, concentration-dependent relaxations evoked by sumatriptan were antagonised by ketanserin and GR127935 (apparent pA2 = 5.9 and 10, respectively). Ketanserin but not GR127935, inhibited Ca(2+)-induced contraction of depolarised strips of guinea-pig ileum longitudinal muscle/myenteric plexus, however, in rabbit saphenous vein and guinea-pig jugular vein, 5-HT receptor mediated responses were insensitive to nifedipine (Ca2+ channel blocker), eliminating the possibility that the inhibitory effects of ketanserin and GR127935 were due to the blockade of voltage-operated Ca2+ channels. Thus, antagonism by ketanserin and GR127935 confirms the presence of 5-HT1D receptors in rabbit saphenous vein and guinea-pig jugular vein. The differential effects of ketanserin and GR127935 on responses elicited by sumatriptan in rabbit saphenous vein and guinea-pig jugular vein may reflect either species differences in 5-HT1D receptors or the involvement of 5 HT1D alpha and 5-HT1D beta receptor subtypes. PMID- 7498312 TI - Autoradiographic localization and characterization of bradykinin receptors in human skin. AB - High affinity [3H]bradykinin binding sites have been identified in human skin cryosections by in vitro autoradiography. Equilibrium binding studies were performed with increasing concentrations of [3H]bradykinin for 120 min in the presence of protease inhibitors at 4 degrees C. In saturation experiments a single class of high affinity binding sites was identified with a dissociation constant Kd of 1.2 +/- 0.8 nM (mean +/- S.E.M., n = 3) and a maximal binding capacity Bmax of 33 +/- 8 fmol [3H]bradykinin specifically bound/mg protein (mean +/- S.E.M., n = 3). Competition experiments revealed a rank order of potency with bradykinin being most effective (bradykinin = [Lys]bradykinin > [Met- Lys]bradykinin > [Tyr]bradykinin > [des-Arg9]bradykinin), whereas [des Arg9]bradykinin was ineffective. This indicates a B2 subtype of bradykinin receptors in normal human skin. Morphological data: autoradiography revealed that bradykinin receptors were localized in the stratum basale of the epidermis. The data are consistent with the hypothesis, that these mitotic active keratinocytes express bradykinin binding sites, that fulfil the pharmacological criteria for true receptors. Diverse stimuli, including bradykinin, play a role in the mediation of cutaneous inflammatory responses (e.g. fluid extravasation, reactive cell proliferation, hyperalgesia). Our data indicate that specific kinin receptors of the stratum basale are likely to contribute to these effects. PMID- 7498313 TI - The sigma receptor ligand JO 1784 (igmesine hydrochloride) is neuroprotective in the gerbil model of global cerebral ischaemia. AB - To assess the effects of the novel sigma receptor ligand JO 1784 ((+)-N cyclopropyl-methyl-N-methyl-1,4-diphenyl-1-yl-but-3-en-1-ylami ne, hydrochloride or igmesine hydrochloride) on behavioural and histological changes following cerebral ischaemia, the gerbil model of cerebral ischaemia was used. Two experiments were carried out. In the first animals were either sham operated, subjected to 5 min of bilateral carotid occlusion or administered JO 1784 (25, 50, 75 or 100 mg/kg p.o.) 1, 24 and 48 h after 5 min bilateral carotid occlusion and histological evaluation carried out 96 h after surgery. In the second experiment the effects of JO 1784 administered at a dose of 100 mg/kg i.p. 30 min, 6, 24 and 48 h post-surgery on home cage activity and nitric oxide (NO) synthase activity in the cortex, hippocampus, cerebellum and brain stem 4 days after surgery was examined. Extensive neuronal death was observed in the CA1 region of 5 min occluded animals. JO 1784 (50, 75 and 100 mg/kg) provided significant protection against this ischaemia-induced cell death (P < 0.03 0.005). In the second experiment a large increase in home cage activity was observed for 5 min occluded animals for 12 h after surgery (P = 0.0018-0.02). A large increase in NO synthase activity was observed in all brain regions for 5 min occluded animals. Post-administration of JO 1784 attenuated the ischaemia induced hyperactivity and increased NO synthase activities.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7498314 TI - Transgenic superoxide dismutase mice differ in opioid-induced analgesia. AB - Autoradiographic data from transgenic mice carrying the human Cu/Zn-superoxide dismutase gene demonstrate an increase in mu-opioid receptor concentration in dopaminergic-related areas and the central grey area. The relative potencies of mu-, delta- and kappa-opioid receptor agonists to induce antinociception in heterozygous and homozygous superoxide dismutase transgenic mice as well as four inbred strains were assessed to determine the functional significance of the increased receptor concentration. Increased superoxide dismutase activity results in an increased sensitivity to mu-agonists in a gene dosage-dependent manner. SOD/Tg/hom mice were less sensitive to the delta-agonist than were SOD/Tg/het mice. The superoxide dismutase transgene did not affect kappa-opioid receptor agonist sensitivity. These data suggest that delta-opioid receptors are not regulated in the same manner as mu-opioid receptors and that kappa-opioid receptors are unaffected by superoxide dismutase activity. PMID- 7498315 TI - Tachykinin NK1 receptors mediate both vasoconstrictor and vasodilator responses in the rabbit isolated jugular vein. AB - We have characterized the receptor(s) mediating contraction and relaxation produced by tachykinins in the rabbit isolated jugular vein. The tachykinin NK1 receptor-selective agonists septide and [Pro9]substance P produced concentration dependent contractions which were potentiated by either the removal of the vascular endothelium (Emax = +106% and +72%, respectively) or by pretreatment with L-nitroarginine (100 microM; 60 min before) (Emax = +123% and +71%, respectively). The tachykinin NK1 receptor-selective antagonist, (+/-)-CP-96,345 ([2-(diphenylmethyl)-N-[(2-methoxyphenyl)-methyl]-1- azabicyclo[2,2,2]octan-3 amine]) (10-300 nM) competitively antagonized septide (pKB = 9.0) with 10-fold greater potency than [Pro9]substance P (pKB = 8.0). In preparations with intact endothelium both septide and [Pro9]substance P (from 0.1 to 100 nM) relaxed the noradrenaline-(10 microM) induced tone, and their effects were markedly reduced by (+/-)-CP-96,345 (100 nM). In noradrenaline-precontracted veins L-nitroarginine (100 microM) reversed the tachykinin-induced vasodilation into a contraction, providing evidence for the involvement of nitric oxide in this response. The tachykinin NK3 and NK2 receptor-selective agonists senktide and [beta Ala8]neurokinin A-(4-10) were either ineffective, or produced small effects antagonized by (+/-)-CP-96,345 (100 nM), respectively. In conclusion, tachykinin NK1 receptors mediate both tachykinin-induced contraction and relaxation in the rabbit jugular vein. This preparation, deprived of the endothelium or pretreated with L-nitroarginine, is suitable for evaluating tachykinin agonists or antagonists. PMID- 7498316 TI - Inhibition of vascular contractions to alpha-adrenoceptor agonists by polymyxin B: impact of heart failure state. AB - In this study, the effects of polymyxin B (a protein kinase C inhibitor) on alpha adrenoceptor stimulation of the canine dorsal pedal artery and saphenous vein were examined. In addition, the question was asked, whether these effects could be altered by the impact of a heart failure state? Blood vessels were obtained at three time points during the development of pacing-induced heart failure in the dog; control (non-paced), 1 week paced and end-stage heart failure. Concentration effect curves were constructed to the alpha-adrenoceptor agonists, namely, noradrenaline and phenylephrine in the absence and presence of polymyxin B (2 x 10(-5) and 10(-4) M). Responses to noradrenaline and phenylephrine were enhanced in the saphenous vein, but not in the dorsal pedal artery, following the onset of heart failure. In the dorsal pedal artery, polymyxin B was found to inhibit the contractions developed to noradrenaline and phenylephrine to a significant degree (P < 0.05) at control and end-stage heart failure. In contrast, in the saphenous vein, polymyxin B inhibited responses developed to noradrenaline and phenylephrine at all time points studied. This inhibition was always more marked against noradrenaline compared to phenylephrine and, similar to the dorsal pedal artery, became more pronounced at end-stage heart failure. Furthermore, the vein was always more sensitive compared to the artery. Interestingly, as heart failure developed, a non-classical broad concentration-effect curve was evident. The high affinity component was more sensitive to inhibition by polymyxin B. This component was absent at end-stage heart failure in response to phenylephrine.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7498317 TI - The pentadecapeptide [Ser1]histogranin impairs passive avoidance learning in mice. AB - The peptides, histogranin and [Ser1]histogranin, were recently shown to modulate NMDA receptor function. In the present study, the effects of intracerebroventricular (i.c.v.) administration of [Ser1]histogranin and of the histogranin receptor antagonist, histogranin-(1-10), were examined on step-down type passive avoidance learning in mice. [Ser1]Histogranin (30-60 nmol) impaired retention, after post-training administration, but not when it was administered just prior to the retention assay. Histogranin-(1-10) (60 nmol) facilitated learning during training, without affecting retention. Co-administration of histogranin-(1-10) with [Ser1]histogranin (60 nmol each) led to a significant prevention of [Ser1]histogranin-induced learning impairment. These results indicate that [Ser1]histogranin impairs passive avoidance learning according to the pattern of NMDA receptor antagonists and involving specific histogranin sites. PMID- 7498318 TI - Nitric oxide synthase activity is elevated in inflammatory lung disease in humans. AB - We have characterised nitric oxide (NO) synthase activity in lung samples from patients with inflammatory lung disease compared to that in normal donor lung. NO synthase activity was measured by the ability of tissue homogenates to convert L arginine to L-citrulline. Higher levels of NO synthase activity were found in samples from patients with inflammatory lung disease (mild asthma, cystic fibrosis, obliterative bronchiolitis after lung transplantation) compared to samples from healthy donors. NO synthase activity in all samples was mainly dependent on the presence of extracellular Ca2+. The increased NO synthase activity in diseased-lung samples suggests a modulatory role for nitric oxide in lung inflammation. PMID- 7498319 TI - Autoradiographic visualisation of [3H]5-carboxamidotryptamine binding sites in the guinea pig and rat brain. AB - We have investigated the distribution of [3H]5-carboxamidotryptamine ([3H]5-CT) binding sites by in vitro autoradiography on sections of guinea-pig and rat brain. In saturation studies, the ligand recognised a saturable, homogeneous population of binding sites with an affinity ranging from 0.19-0.45 nM depending on the region. The labelling pattern was heterogeneous, and the displacement pattern with different competing drugs selective for different 5-HT receptor subtypes was complex. [3H]5-CT appeared to label 5-HT1B/5-HT1D sites in the substantia nigra, globus pallidus and caudate/putamen, as the binding in these regions was displaced by the 5-HT1B/1D receptor selective agents sumatriptan, CP 122,288 and GR-127,935. In the hippocampus and lateral septum, the very dense [3H]5-CT binding was displaced with high affinity by the 5-HT1A receptor selective agonist 8-hydroxy-dipropylaminotetralin ((+/-)-8-OH-DPAT), dihydroergotamine and 5-HT. In contrast the affinity of the 5-HT1 receptor antagonists spiperone and methiothepine was much lower than their previously published potency at 5-HT1A receptors. The affinity of agonists, taken together with the fact that the distribution of these [3H]5-CT sites overlaps that of [3H]8-OH-DPAT binding sites in serial sections, suggest that these sites correspond to 5-HT1A receptors. Their atypical properties deserve further investigations. While [3H]5-CT binding at 5-HT1B/1D sites and these atypical 5 HT1A sites was inhibited by the GTP analogue 5'-beta, gamma-imidotriphosphate, [3H]5-CT binding in the superficial cortical layers and in midline thalamic nuclei was insensitive to this agent. It was however displaced by low concentrations of spiperone, clozapine and methiothepine, but not by sumatriptan, CP-122,288, GR-127,935 or dihydroergotamine. This binding profile is similar to that of 5-HT7 receptors, while the spatial distribution of these sites matches the known distribution of 5-HT7 messenger RNA. We did not find evidence of [3H]5 CT labelling to 5-HT5 receptors, in spite of their reported high affinity for this ligand. It is concluded that [3H]5-CT, in the presence of selective blockers, can be used to investigate the properties of 5-HT1A, 5-HT1B/1D and 5 HT7 receptors in the rodent brain, although further studies are required to explain the atypical features of [3H]5-CT binding in 5-HT1A receptors containing regions. PMID- 7498320 TI - Association of microvascular leakage with induction of nitric oxide synthase: effects of nitric oxide synthase inhibitors in various organs. AB - Endotoxin (Escherichia coli lipopolysaccharide 0111:B4, 3 mg/kg i.v.) induced the expression of a calcium-independent nitric oxide (NO) synthase, determined after 5 h in cardiac, hepatic, pulmonary and renal tissues, as assessed by the conversion of radiolabelled L-arginine to L-citrulline. This widespread induction of NO synthase in these conscious rats was associated with microvascular injury, as assessed by the vascular leakage of radiolabelled human serum albumin. Concurrent administration of the NO synthase inhibitor. NG-nitro-L-arginine methyl ester (L-NAME, 1-5 mg/kg s.c.) with endotoxin, provoked acute vascular leakage within 2 h in the various organs. By contrast, the delayed injection of L NAME (1-5 mg/kg s.c.) or NG-monomethyl-L-arginine (12.5-50 mg/kg s.c.) until 3 h after endotoxin challenge inhibited the subsequent microvascular leakage in these organs. These effects of NO synthase inhibitors were reversed by L-arginine (300 mg/kg s.c.) pretreatment. These results support a protective role of constitutive NO synthase in the early phase of endotoxin shock. Such actions contrast with the aggressive actions of the products of inducible NO synthase in the development of widespread microvascular injury in endotoxemic states. PMID- 7498321 TI - Differential effects of classical and newer antipsychotics on the hypermotility induced by two dose levels of D-amphetamine. AB - The inhibitory effects of a variety of established and putative antipsychotic compounds on the hypermotility induced by D-amphetamine at two dose levels (0.5 and 2.0 mg/kg) have been studied. Classical antipsychotics (haloperidol, fluphenazine and cis(Z)-flupentixol) and the selective dopamine D2 receptor antagonist remoxipride inhibit hypermotility in the two conditions with similar potencies, whereas sertindole, clozapine, risperidone, ziprasidone and olanzapine preferentially inhibit the effect of the low dose of D-amphetamine (selectivity ratios between 6.5 and 18). Seroquel, amperozide and the selective 5-HT2A receptor antagonist MDL 100.151 ((+/-)-alpha-(2,3-dimethoxyphenyl)-1-[2-(4 fluorophenylethyl)]-4-p iperidine - methanol) have no effect on D-amphetamine 2.0 mg/kg, but inhibit the response to D-amphetamine 0.5 mg/kg. The alpha 1 adrenoceptor antagonist prazosin inhibits the motility response to D-amphetamine 0.5 mg/kg with slightly higher potency than that to D-amphetamine 2.0 mg/kg, whereas the 5-HT2A/2C receptor antagonist ritanserin selectively inhibits the effect of D-amphetamine 0.5 mg/kg. The histamine H1 receptor antagonist mepyramine is ineffective in both models. All compounds, except remoxipride, MDL 100.151 and ritanserin (which are ineffective) inhibit spontaneous locomotor activity at dose levels close to those inhibiting the response to D-amphetamine 2.0 mg/kg. Prazosin has partial inhibitory effect. In conclusion, dopamine antagonism has similar inhibitory effect on hyperactivity induced by low and high D-amphetamine dosages, alpha 1-adrenoceptor antagonism also contributes to both effects, whereas 5-HT2 receptor antagonism selectively interacts with the low D amphetamine dose. This indicates that the responses to D-amphetamine 0.5 and 2.0 mg/kg are differently modulated by these neurotransmitters.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7498322 TI - Pharmacology of a potent long-acting imidazole-5-acrylic acid angiotensin AT1 receptor antagonist. AB - The angiotensin II antagonistic activity of SB 203220, [E-alpha-[[2-butyl-1-(4 carboxy-1-naphthalenyl)methyl]-1H- imidazol-5-yl]-methylene]-2-thiophene-propanic acid], was examined in several in vitro and in vivo assays. SB 203220 displaced [125I]angiotensin II binding from a variety of tissues including the cloned human AT1 receptor (IC(50)5-15 nM). SB 203220 (10 microM) did not interact with AT2, endothelin (ETA and ETB) or calcitonin gene-related peptide receptors. [3H]SB 203220 bound with high affinity to the AT1 receptor (Kd = 4.9 nM), but dissociated from the receptor at a much slower rate when compared to [3H]SK&F 108566. SB 203220 antagonized intracellular Ca2+ mobilization induced by angiotensin II in rat vascular smooth muscle cells and exhibited a selective and partially insurmountable antagonism of angiotensin II-induced contraction in isolated rabbit aorta. In the aorta, SB 203220 produced a concentration-dependent parallel shift in the concentration-response curve to angiotensin II [EC30 = 5.94 +/- 1.6 10(-11) M] and depressed the maximal contractile response to angiotensin II by approximately 35%. The antagonistic effect of SB 203220 in rabbit aorta was slowly reversible compared to SK&F 108566. SB 203220 displayed no agonist activity and had no effect on the contractile responses to KCl, endothelin-1 or norepinephrine. In rats, SB 203220 at 10 mg/kg i.v. inhibited angiotensin II induced aldosterone release. Intraduodenal or oral administration of SB 203220 (1 10 mg/kg) to conscious rats and dogs inhibited the pressor responses to exogenous angiotensin II.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7498323 TI - Functional characterization of the prostanoid DP receptor in human myometrium. AB - Spontaneous contractile activity of strips of human myometrium obtained from non pregnant donors at the time of hysterectomy was inhibited by the selective prostanoid DP receptor agonists BW 245C (5-(6-carboxyhexyl)-1-(3-cyclohexyl-3 hydroxypropyl)hydantoin) and ZK110841 ((5Z,13E)-(9R,11R,15S)-9 beta-chlor-15 cyclohexyl-11,15-dihydroxy-16,17,18,19, 20-pentanor-5,13-prostadienoic acid) with pEC50 values of 8.4 and 7.3 respectively but prostaglandin D2 produced a biphasic effect. In the presence of the TP receptor antagonist L670596 ((-)-6,8-difluoro-9 p-methylsulfonyl benzyl-1,2,3,4-tetrahydrocarbazol-1-yl-acetic acid), contractile activity induced by the FP receptor agonist, cloprostenol ([1R-[1 alpha(Z),2 beta(1E,3R),3 alpha,5 alpha]]-7-[2-[4-(3- chlorophenoxy)-3-hydroxy-7-butenyl]-3,5 dihydroxycyclopentyl]-5-he ptenoic acid), was inhibited by BW 245C (pEC50 = 7.5), ZK110841 (pEC50 = 6.7) and prostaglandin D2 (pEC50 = 6.3). Under these conditions both prostaglandin J2 and 9 alpha,11 beta-prostaglandin F2 were inhibitory partial agonists. All compounds were antagonized by the selective DP receptor antagonist BW A868C (3-benzyl-5-(6-carboxyhexyl)-1-(2-cyclohexyl-2 hydroxyethylamino)h ydantoin), but the pKB values were both concentration dependent (pKB versus BW 245C at 10 nM = 9.1, at 50 nM = 8.3) and agonist dependent (pKB at 10 nM versus BW 245C = 9.1, versus ZK110841 = 7.4). Both agonist and antagonist potencies support the existence of DP receptors in human myometrium. The concentration and agonist dependence of the action of BW A868C suggests that putative DP receptor agonists relax human myometrium by more than one mechanism. These observations may be explained by the existence of subtypes of DP receptor in human myometrium. PMID- 7498324 TI - Local anesthetics reduce the inhibitory neurotransmitter-induced current in dissociated hippocampal neurons of the rat. AB - The effects of local anesthetics on amino acid-induced currents were examined using the whole-cell configuration of the patch clamp technique in dissociated hippocampal pyramidal neurons of the rat. Lidocaine (3 mM) decreased the glycine induced Cl- current (Gly-ICl) more potently (to 46% of the control value) than the gamma-aminobutyric acid-induced Cl- current (GABA-ICl; to 75%), whereas the agent had little effect on the excitatory glutamate response. The reduction in the Gly-ICl was dose-dependent, with a dissociation constant (KD) of 3 mM and a Hill coefficient of 0.96. A non-competitive inhibition was suggested by a double reciprocal plot of the effects of lidocaine on the concentration-response curve of the Gly-ICl. Benzocaine, a neutral local anesthetic at physiological pH, decreased the Gly-ICl more potently than lidocaine, while QX314, a permanently charged quaternary derivative of lidocaine, produced a much smaller inhibition, thereby indicating that the neutral form of local anesthetics is more effective in reducing the Gly-ICl. The depression of the Gly-ICl and GABA-ICl in central neurons may contribute to local anesthetic-induced convulsions. PMID- 7498325 TI - Endogenous corticosteroids mediate the neutrophilia caused by platelet-activating factor in the mouse. AB - Platelet-activating factor (PAF; 100 ng i.v.) transiently modified the number of circulating neutrophils in the mouse, inducing a fast neutropenia (2 min) followed by a late onset neutrophilia (2 h). The potential involvement in PAF induced neutrophilia of granulocytotic agents such as interleukin-1 and tumor necrosis factor-alpha could be excluded on the basis of the ineffectiveness of interleukin-1 receptor antagonist and of a specific monoclonal antibody anti murine tumor necrosis factor-alpha. PAF granulocytosis was preceded by a significant rise in plasma corticosterone at 20 min. The involvement of endogenous corticosteroids was confirmed by the experiments with adrenalectomized mice and in animals pretreated with the steroid antagonist RU486 (11 beta-(4 dimethyl amino-phenyl) 17 beta-hydroxy, 17 alpha(prop-1-ynyl) estra 4,9-dien-3 one), where PAF-induced neutrophilia was greatly reduced (approximately 50%). Moreover, sustained increase in plasma corticosterone by administration of adrenocorticotropic hormone was paralleled by an intense neutrophilia. We show evidence that endogenous corticosterone acts through the glucocorticoid-inducible protein lipocortin 1. PMID- 7498326 TI - Effect of antiarrhythmics on the release of adenosine in rat hearts with coronary occlusion and reperfusion. AB - In isolated perfused rat hearts the left coronary artery was occluded for 5 min, with subsequent reperfusion for 20 min. During the reperfusion severe tachyarrhythmias were observed, with ventricular fibrillation occurring in all hearts. Simultaneously, large amounts of adenosine and its degradation products inosine, hypoxanthine, xanthine and uric acid were released into the coronary perfusate. The antiarrhythmics quinidine, lidocaine and gallopamil significantly decreased the release. The effect of quinidine and lidocaine was linked with the antifibrillatory action of these drugs. Also the interruption of fibrillation immediately after its appearance by potassium chloride decreased the release of adenosine and its metabolites in a highly significant way. The effect of gallopamil on the release was independent of an antifibrillatory action. The findings indicate that different kinds of antiarrhythmic drugs can affect the release of nucleosides and oxypurines in hearts with ischaemia and reperfusion. PMID- 7498327 TI - Norepinephrine loss exacerbates methamphetamine-induced striatal dopamine depletion in mice. AB - Evidence is accumulating that norepinephrine depletion enhances the neurotoxic effect of the parkinsonism inducing neurotoxin 1-methyl-4-phenyl-1,2,3,6 tetrahydropyridine (MPTP). In this study we investigated whether norepinephrine loss potentiates methamphetamine-induced striatal dopamine depletion. Injection of C57BL/6N mice with methamphetamine (2 x 5 mg/kg i.p., at 2-h intervals) produced only a partial (50%) striatal dopamine depletion 7 days after drug administration. Pretreatment with the selective noradrenergic neurotoxin N-(-2 chloroethyl)-N-ethyl-2-bromobenzylamine (DSP-4; 50 mg/kg i.p.) enhanced methamphetamine-induced striatal dopamine depletion by 86%, without decreasing striatal dopamine levels when injected alone. Our results extend previous findings obtained with the dopaminergic neurotoxin 1-methyl-4-phenyl-1,2,3,6 tetra-hydropyridine in DSP-4-pretreated mice. PMID- 7498328 TI - [Morphological studies on preputial gland of mice--1. Light and transmission electron microscopic observations of preputial gland in male mice]. AB - Male BALB/c mice isolated after weaning were housed two mice per cage at the age of 10 weeks. After 1 week, each pair of mice was observed for social dominance, and compared with control mice kept in isolation. The preputial gland in the socially dominant animal was histologically examined. The preputial gland consisted of a congregation of branched simple tubuloacinar gland and typical holocrine secretion on morphological examination. Acinar cells in the dominant animals were large, multiplicative and hypertrophic compared with those of controls and subordinates. Dominant animals required copious secretion for attainment and maintenance of a dominant social position. Acinar cells of subordinates were very small, flattened and atrophied because fatty secretion particles filled the luminal duct. These accumulated secretion particles pressed the acinar cells and caused degeneration. Not only luminal acinar cells but also central acinar cells and some basal acinar cells were degenerated. Electron micrographs showed that basal acinar cells in dominant animals contained many mitochondria, agranular endoplasmic reticulum and free ribosomes in the cytoplasm, whereas those of subordinates contained myelin bodies and lipid droplets. Therefore, it is suggested that the synthesis of secretion particles in acinar cells of subordinates hardly occurred. PMID- 7498329 TI - [Age-related changes in the retina of WBN/Kob rats--a pathological study]. AB - Male rats of the WBN/Kob strain, which are known to spontaneously develop diabetes with aging, were examined for histopathological changes in the retina. Five rats (10 eyes) each of WBN/Kob and Wistar/ST as a control were used, and the thickness of the retinal layers, both the central region and the peripheral region of the retina, were measured on weeks 1, 2, 3, 5, 9, 13, 17, 23, 27, 36, 45, 54, 67 and 80 after birth. The rod and cone cell layer in WBN/Kob rats was under-grown, and its thickness decreased 71.7% in the central zone and 59.3% in the peripheral zone of the retina compared with that of the control. In the central and peripheral retina, the rod and cone cell layer, outer nuclear layer and outer plexiform layer, as well as the inner plexiform layer in the central retina, gradually decreased in thickness from 5-45 weeks of age. In the central and peripheral parts of the retina, the number of nuclei decreased in accordance with the thinning of the nuclear layer. The thinned layers showed only cell loss. The rod and cone cell layer of the peripheral retina was thinner than that of the central retina. We obtained the following findings in the retina of male WBN/Kob rats. First, the rod and cone cell layer is undergrown compared with that of the control. Second, the first change occurred in the rod and cone cell layer. Third, the thinning of the rod and cone cell layer appeared at 5 weeks of age, and thinning with aging was slow. And finally, the thinning of the peripheral retina was more severe than that of the central retina. From the above findings, it seems that retinal changes in WBN/Kob rats are similar to the retinal degeneration of rds mice (retinal degeneration slow mice) and that WBN/Kob rats provide a useful animal model for human retinopathy. PMID- 7498330 TI - [Histological studies on postnatal development of the musk gland in house musk shrews (Suncus murinus)]. AB - The development of the musk gland in house musk shrews aged 0 to 21 days was studied lectin histochemically. On about 3 days of age, the musk gland could be discriminated from surrounding skin. This organ developed markedly on days 3 to 7, and its development continued to 21 days of age. Histologically, on day 1, the bulges that should develop into musk glands were identified in the vicinity of hair germs. On days 3 to 7, these bulges developed markedly, and on day 21 their structure revealed almost the same morphology with that in mature musk glands. We used 7 lectins; ConA, RCA I, PNA, SBA, UEA-I, DBA, and WGA. On day 1, 6 lectins except for WGA labelled the cell of the musk gland. WGA began to label it on day 3. Until 7 days of age, lectins bound to only the cytoplasm of the cell of musk glands. On days 12, however, PNA and UEA-1 labelled the cell membrane equally, and RCA I labeled it on day 21. PMID- 7498331 TI - [Age-related changes and sex differences on the serum chemistry values in Sprague Dawley rats--I. 6-30 weeks of age]. AB - Age-related changes of 27 items in serum chemistry were investigated in Sprague Dawley rats of both sexes from 6 to 30 weeks of age. The following 12 items were shown as an increase in those values during growth and maturity, i.e., total protein, albumin (female only), glucose, total cholesterol, triglyceride, phospholipid (female only), beta-lipoprotein, cholinesterase (female only), asparate aminotransferase (female only), creatinine, direct-bilirubin and total bilirubin. However, the following 4 items decreased with aging, i.e., asparate aminotransferase, alkaline phosphatase, creatine phosphokinase and inorganic phosphorus. No age-related changes were found in the values for calcium, sodium and chloride in both sexes and for alanine aminotransferase, cholinesterase and albumin in males. The sex differences were shown in the following 12 items: higher values in males were alkaline phosphatase, creatinephosphokinase, glucose and inorganic phosphorus, and higher values in females were cholinesterase, albumin, phospholipid, non-esterified fatty acid, urea nitrogen, direct bilirubin, total-bilirubin and serum iron. No sex-related differences were found in the values for calcium, sodium, chloride and total cholesterol. PMID- 7498332 TI - [Fate of two different stages of embryos transferred to an identical recipient in mice]. AB - To define the developmental fate of two different stages of embryos existing together in the mouse reproductive tract, different stages of fresh and frozen thawed embryos were transferred separately into the oviducts of identical recipients. ICR and FvB embryos were flushed from the oviducts and/or uterus of superovulated females on Day 1 (2-4-cell stage) and Day 2 (8-cell-morula stage) of pregnancy. Day 1 embryos were transferred separately to the right or left oviduct of recipients, while the other oviduct received Day 2 embryos. There were no significant differences between Day 1 and Day 2 embryos with respect to the number of implantation sites and live fetuses in either the fresh or frozen thawed embryos. These results concerning to the embryonic stage were similar to those of control experiments, in which Day 1 or Day 2 embryos were transferred to both right and left oviducts of recipients. Furthermore, no strain differences were observed in this study. No developmental retardation or anomalies were observed in fetuses derived from either Day 1 or Day 2 embryos. The embryo transfer in this study revealed that differences in developmental stage at preimplantation were synchronized by the maternal uterine environment. PMID- 7498333 TI - [Morphology of the mandibular condyle in osteopetrotic (op/op) mouse]. AB - Osteopetrosis is an inherited metabolic disease which produces an excessive accumulation of bone caused by deficiency of osteoclasts. It is characterized by systemic bone sclerosis, cranial deformation and failure of tooth eruption due to defective bone resorption. In this study, morphological change of the mandible was observed in growing osteopetrotic (op/op) mutant mice. Deformation of the condylar process, non-ossification of the condyle head and underdevelopment of the lateral pterygoid muscle were observed in the mutant mice. Although these phenomena were brought about by failure of the bone remodeling, it was simultaneously suggested that the ossification of the condyle head concerned with the mastication. PMID- 7498334 TI - [Morphological study on the gastric mucosa in diabetes mellitus rats induced by streptozotocin]. AB - To clarify the pathogenesis of vulnerability to acute gastric mucosal lesion (AGML) in diabetes mellitus (DM), we investigated histopathologically the gastric mucosa of the fundic gland in streptozotocin-induced DM rats. The length of the mucosa and the thickness of the surface epithelial cell (SEC) layer were measured, and the number of proliferating cell nuclear antigen (PCNA) positive cells was counted. As results, compared with control rats, the ratio of the length of SEC layer to that of mucosa, and the number of PCNA positive cells were significantly decreased in DM rats, whereas the length of the mucosa tended to extend in DM rats. Consequently, it is considered that attenuation of mucosal barrier in the SEC layer can be closely associated with susceptibility to occurrence of AGML in DM. PMID- 7498335 TI - [Age-related memory impairment and hippocampal damage in ddY male mice]. AB - The relationship between behavioral change and hippocampal lesion was studied in ddY male mice, at 4.5, 20, 40 and 60 weeks of age. In passive avoidance response, the aging mice (40 and 60 weeks of age) showed shorter latency than young mice (4.5 weeks of age). The degenerated pyramidal cells were more numerous after 40 weeks of age in the CA3 than CA1. The mean incidence of the degenerated pyramidal cells in the CA3 was 20.8% at 60 weeks of age. In 71.9% of mice, impairment of passive avoidance response was associated well with hippocampal lesions. These results indicate that the hippocampus plays some role in the memory in mice. The ddY mice may be possible to use as models for reversible hippocampal lesion. PMID- 7498336 TI - [Invention of a forced-air-ventilated micro-isolation cage and rack system- environment within cages: temperature and ammonia concentration]. AB - There is little information about microenvironments within laboratory animal cages, although several guidelines on the macroenvironment in laboratory animal rooms have been reported. We have developed a forced-air ventilation system (FVMIS) to improve laboratory animal well-being. In this study, we tested the performance of the FVMIS in terms of temperature and ammonia concentration within cages. Continuous measurement of temperature and ammonia concentration was carried out in FVMIS cages housing 5 rats each. The average temperature within the cages was 22 degrees C to 24 degrees C, and the temperature difference among cages was 0.93 degrees C. The air exchange rate used was 65 changes per hour. FVMIS was capable of controlling the temperature within cages uniformly and accurately with a lower air exchange rate than with the conventional system. The ammonia concentration reached 20 ppm within 9 days in a filtercapped conventional cage, but took 13 to 16 days to reach 20 ppm with FVMIS. It appears that FVMIS is capable of suppressing the ammonia concentration increase in cages, thereby reducing the frequency of cage changes and the stress to laboratory animals when cages are changed. In the present study, it was confirmed that FVMIS contributes not only to microenvironmental control in terms of temperature and ammonia concentration with a lower air exchange rate, but also to animal well-being. PMID- 7498337 TI - [Histopathological studies on senile plaques and cerebral amyloid angiopathy in aged cynomolgus monkeys]. AB - The incidence and morphological characteristics of senile plaque and amyloid angiopathy in the cerebrum of six aged cynomolgus monkeys (Macaca fascicularis), 20 to 29 years old, were studied histopathologically and immunohistochemically. By periodic acid methenamine silver stain (PAM) and alkaline Congo red stain, senile plaques were detected in 5 out of 6 cases, and 3 of them were positive for amyloid in the wall of capillaries and arterioles in the cerebral cortex. Senile plaques were classified into three types. Mature plaques, including classical and primitive types, were more frequently observed than the immature diffuse type. Senile plaques were often seen in the cortex of temporal lobe, putamen and head of caudate nucleus. Since mature types of senile plaques were seen frequently around vascular amyloid deposition and no amyloid angiopathy was detected in the areas without senile plaques, the close relation between senile plaque of the mature type and amyloid angiopathy might be considered. All senile plaques and amyloid angiopathy were positively stained immunohistochemically with antibody against amyloid beta-protein (A beta P) 1-40 synthetic peptide, but all diffuse and some primitive plaques were negative for antibody against A beta P 8-17 synthetic peptide. Neither senile plaque nor amyloid angiopathy was detected in the cerebrum of 15 young monkeys, 9 to 11 years old, examined as controls. PMID- 7498338 TI - [Study of biochemical gene markers in the Mongolian gerbil (Meriones unguiculatus)]. AB - The present study was conducted to analyze biochemical marker genes and to clarify genetic variation in 4 Mongolian gerbil (Meriones unguiculatus) strains (MGS/Sea, MON/Jms Gbs, Kwl: Mongolian Gerbil and Hos: Mongolian gerbil) maintained in Japan. Cellulose acetate, starch gel or agar gel electrophoresis was used. Of the 23 kinds of proteins detected, variation was found only in liver acid phosphatase (Acp2); no variation in other proteins was found within or among strains. The results suggest that these 4 Mongolian gerbil strains are genetically similar. A strain difference was found in the Acp2 locus of liver acid phosphatase. The Kwl: Mongolian Gerbil strain had a band of Acp2 locus, whereas MGS/Sea, MON/Jms and Hos: Mongolian gerbil strains did not. PMID- 7498340 TI - [Effects of feed and mating age on reproductive performance in Syrian hamsters]. AB - In order to utilize Syrian hamsters in selection experiments, we investigated desirable conditions of feed and mating age for reproduction in females. Hamsters were subdivided into two groups, raised on pellet feed for herbivores (ZC-2) and breeding (MB-1), respectively. Half of the hamsters in each group were switched to the other feed, respectively, after mating. They were mated at 8 or 12 weeks of age. In the group that has been fed on ZC-2 before mating and MB-1 after mating, there was significantly more females giving birth and weaning, and exhibited the largest litter size and litter weight at 3 weeks of age (P < 0.01). The hamsters of the groups mated at 12 weeks of age exhibited a higher percentage of females giving birth and litter size and litter weight at birth than those at 8 weeks of age, but that was no significant difference in litter size and litter weight at 3 weeks of age. We conclude that Syrian hamsters raised by fibrous feed before mating gave optimal reproductive performance. PMID- 7498339 TI - [Mixed lymphocyte reaction among goat strains fixed by DRB/DQB haplotype in the MHC class II region]. AB - One-way mixed lymphocyte reaction (MLR) tests were carried out on three strains of Korean native goat, A, D and E, which had been fixed by DRB/DQB haplotype in the major histocompatibility complex (MHC) class II region. In the case of MLR combined within the same strain, the values for the stimulation index (S.I.) were from 1.08 +/- 1.26 (within the D strain) to 3.19 +/- 2.28 (within the A strain). On the other hand, in the case of MLR combined for the different strains, the S.I. values were from 10.47 +/- 4.31 (E strain as the stimulator and A strain as the responder) to 36.08 +/- 7.42 (D strain as the stimulator and E strain as the responder). The difference between the S.I. values for the same and different strains was clearly significant (p < 0.001). To elucidate the effect of a combination of the DRB/DQB haplotype on the S.I. values, A and E strain and AE heterozygote animals were used in the MLR tests. With MLR combined heterozygote as the stimulater and homozygote as the responder (A or E strain), the mean S.I. value was 9.62 +/- 3.9. This value showed high levels in the different strains, but showed a significant difference from that of the same strain (p < 0.001). On the other hand, with MLR combined homozygote as the responder (A or E strain) and a heterozygote as the stimulater, the mean S.I. value was 1.46 +/- 1.17. This value showed low levels as well as within the same strain, but showed a significant difference from that of different strains (P < 0.001).(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7498341 TI - [A biological study of inbred Weiser-Maples guinea pigs--urinalysis, hematological and blood-chemical values and organ weights]. AB - Urinalysis, hematological and blood chemical examinations and measurement of organ weights were carried out for the purpose of collecting background data on Weiser-Maples (WM) which was established as an inbred strain of guinea pigs. The results were compared with those for the commercial guinea pigs, Std: Hartley (H). In addition, the measurement methods for urinary specific gravity and whether centrifugation of urine influences the results of urinalysis or not were examined. The specific gravity values calculated from the volume and weight of urine were highly correlated with those from the refraction rate. The results of urinalysis of the supernatant of centrifuged urine were similar to those of urine not centrifuged, but the results suggested that the use of the supernatant provided a more accurate measurement of urine in guinea pigs. The positive rate for urinary protein was higher in the WM strain (38.9%) than in the H strain (0%). Although the WM strain showed higher or lower values than the H strain in some items of hematological and blood chemical examinations and organ weights, we could not regard them as characteristic of the WM strain. PMID- 7498342 TI - [Effects of the long-term administration of methamphetamine on body weight, food intake, blood biochemistry and estrous cycle in rats]. AB - We have a big problem with the abuse of amphetamine and its close relative, methamphetamine (MAP) in Japan. As an animal model of people who abuse MAP, male and female rats were treated with MAP (0.1-10.0 mg/kg/day) for a long time. The results obtained in the present study were as follows. 1. Body weights in MAP treated groups showed a dose-dependent decrease with loss of food intake. 2. Food intake in rats treated with MAP decreased, compared with the control, but when treatment with MAP was discontinued, food intake increased dramatically. 3. In a blood biochemistry assay, the turnover of protein and lipid was suppressed in rats after MAP. 4. The administration of MAP appeared to disturb the estrous cycle in female rats. PMID- 7498343 TI - [Spontaneous tumors in the Mongolian gerbil (Meriones unguiculatus)]. AB - The post mortem examination of 158 Mongolian gerbils (80 males and 78 females) revealed that 61 cases (38.6%) of them carried spontaneous neoplasms [22/80 (27.5%) of males and 39/78 (50%) of females]. In the male, sebaceous gland carcinoma at the abdominal skin was observed in 18/22 (81.8%) of tumor cases. In the female, granulosa cell tumor was seen in 31/39 (79.4%) of tumor cases. These two tumor types were markedly predominant in gerbils older than 3 years. The each incidence of other types of tumor was less than 5% in male and female animals. PMID- 7498344 TI - [Experimental pulmonary metastasis of Chinese hamster mesenchymal chondrosarcoma cell lines]. AB - Pulmonary metastases of chinese hamster mesenchymal chondrosarcoma cell lines, MCS-1 (undifferentiated type) and MCS-8 (differentiated type), were examined by intravenous transplantation into athymic nude mice. The incidence of pulmonary metastasis of MCS-1 was 100% and that of MCS-8 was 33% at the 23rd day after transfer. Mean number of metastatic nodules in the lung was 41 in the former and only 3 in the latter. Mean survival time of mice with MCS-1 injection (5 x 10(4) cells) was 27 days and that with MCS-8 (5 x 10(4) cells) was 48 days after transfer. At the 42nd day after transfer of MCS-8, however, the incidence of pulmonary metastasis was 100%. These data suggest that the tumor growth rate in the metastatic lesion, as well as the affinity to the target organ, is very important for evaluation of experimental metastasis. PMID- 7498345 TI - [Behavioral characteristics of open-field activities in NOD mice]. AB - The open-field test was carried out on NOD mice of both sexes between the ages of 40 to 300 days. The behavior of both sexes of mouse was observed for 3-minute periods with an interval of 20 days. The mean scores of ambulation and rearing increased at the ages of 40-80 days old, and both scores decreased more than 100 days old. The onset of overt diabetes (DM) in female mice were observed after 120 days old. After the onset of DM with great loss of body weight, the scores of ambulation and rearing were dramatically decreased. Other parameters, urination, defecation, and preening were not changed with age of both sexes. These data suggest that conspicuous weight loss with the expression of overt diabetes in NOD female mouse might correlate to decrease of the ambulation and rearing scores. PMID- 7498346 TI - [Brain deformation caused by hyperplasia molar teeth (macrodonts) in the Japanese field vole (Microtus montebelli)]. AB - Protrusion of hyperplastic upper molar teeth (macrodonts) into the cranial cavity was observed in the Japanese field vole. Protrusions of the upper molars occurred on both sides, but were large on the left than on the right. The teeth invaded the cranial cavity and brain parenchyma. Large concavities on the ventral side of the brain were interposed between the lateral olfactory tract and the optic tract. Invation by one macrodont reached the basal nuclei, hypothalamic and thalamic nuclei. The brain was deformed bilaterally, and nearly the entire olfactory bulb, globus pallidus, internal capsule, and lateral preoptic nucleus, and a portion of the hypothalamus, thalamus and anterior commissure were absent on the left side. PMID- 7498348 TI - [The evolution of etiological research on tumors]. PMID- 7498347 TI - [Biological characteristics of Korean native goats]. PMID- 7498349 TI - [The ethical aspects of human experimentation]. PMID- 7498350 TI - [Epidemiology and territory: what structuring? A local experience in the Lombard region]. PMID- 7498351 TI - [DRGs and MDCs in the evaluation of surgical departments: the experience of Local Health Screening Unit No. 13 of the Veneto Region]. AB - This paper discusses the utilization of MDC and DRG for the evaluation of the activities of surgical divisions. Discharge data on 1901 admissions of the second semester 1992 (97.1% of admissions) to three surgical divisions of ULSS 13 in Veneto Region were assigned to HCFA-DRG, 6th version: 423 (22.3%) in division A; 681 (35.8%) in B; 797 (41.9%) in C. MDC 6 (Diseases and Disorders of the Digestive System) was the most frequent MDC in every division; MDC 4 (Diseases and Disorders of the Respiratory System) had an high proportion only in division A, showing the peculiar feature of this division that effects the thoracic surgery of the whole ULSS. Division A had the highest proportion (56.3%) of admissions for surgical DRGs; division C the lowest (35.4%). DRG 119 (Vein Ligation and stripping) was the most frequent surgical DRG in every division: in the whole sample its DRG specific pre-surgical average length of stay was 1.7 days, and the total average length of stay was 4.7 days. MDC and DRG were usefull in describing case-mix, but it is fundamental to have an appropriate knowledge of these instruments, to avoid in reaching inappropriate considerations, because it was inappropriate the use of the same instruments. PMID- 7498352 TI - [Indirect morbidity indicators and social differences in the ISTAT quarterly workforce survey]. AB - Information on the health condition job related of the population are often collected in specific studies, usually limited over time and concerned with a small part of the general population. Looking at official statistics, sources such as national health surveys (in Europe and in North America) are rich on health data, but often offer little information on job-conditions, and job history. On the other hand, one also can find official sources, mainly concerned with labour and job related topics, in which information related to the health status of the people interviewed are routinely collected. These are the Labour Force Surveys, carried out in the most of developed countries. In the Italian Labour Force Survey (RTFL) the Italian Statistical Institute (ISTAT) quarterly collects information on health-related sick absence, part-time job and unemployment. Data from RTFL of two Italian regions (Lombardy: 1985, '86, '89 and Veneto: 1985, '86) show differentials by age and job-position that are consistent with expectation, encouraging an extensive use of this source for surveillance purposes at the national and regional level. PMID- 7498353 TI - [The validity of the certification of natimortality: the experience of a local study]. AB - Although several study confirmed the inaccuracy of information in death certificates, very few investigations were performed to assess the validity of stillbirth certificate reporting. The aim of the present study was to evaluate the extent to which the clinical diagnosis of stillbirth cause reported in certificates can be considered sensitive and specific. Eighty-five consecutive stillbirths were independently classified by two neonatologists with certificate information, using the new Wigglesworth classification, and the reliability of the classification process was evaluated. Afterwards we compared the classes made with certificate causes and the ones made with clinical-autoptic data (the "gold standard") and we estimated sensitivity and specificity of certificate cause of stillbirth. The reliability of the classification process was very high. Values of sensitivity were considered unacceptable, mainly in the field of congenital malformations, intra-partum asphyxia and other specific causes. By contrast, specificity was found to be satisfactory. The accuracy of clinical stillbirth diagnosis is not sufficient and good description needs the contribute of post mortem necropsy. PMID- 7498354 TI - [Tobacco smoking among the students of professional nursing schools in the province of Bologna]. AB - A study into the smoking habits of the Nursing School students of Bologna Province was carried out. 1077 (97.8%) of all students answered the self administered questionnaire and 44.6% of them were current or occasional smokers. 37.2% of males and 31.1% of females were current smokers. Overall nurse students smoked in a greater proportion in respect of coecal young people and females were generally lighter smokers and smokers of light cigarettes. Age, smoking habits of parents, siblings and partners were positively associated with smoking status of students while sex, civil status, education birth area, sport practice and parents profession did not show any significant association. PMID- 7498355 TI - [The knowledge, attitudes and practice in regard to the risk for occupational HIV infection in a group of gynecologists from 3 Italian regions]. AB - The risk of occupational infection by blood-borne pathogens has been well evaluated. However, a low perception of this risk among health care workers (HCWs) and a low compliance to infection control measures has been documented by several studies. During a meeting of gynecologists of Southern Italy, a survey of knowledge, attitudes and practices (K.A.P.) on HIV and other blood-borne viral infections was conducted among 143 obstetricians-gynecologists. 133 (93%) HCWs answered the questionnaire. 81% of them recalled almost one occupational exposure during their career and 54% almost two in the past year. Our data suggest a low knowledge about HIV and the other blood-borne viral infections and Universal Precautions (UP): only 16% of HCWs knew the rate of vertical transmission of HIV, less than 15% knew the rate of seroconversion after occupational exposure to HBV and HCV, and finally only 33% knew to which body fluids apply UP. Moreover, 93.7% of HCWs believed that HIV antibody screening of all patients is an effective strategy to reduce the risk of occupation HIV infection. More than 90% of gynecologists used to request HIV-antibodies to pregnant women, 21% after the first trimester. Although HCWs interviewed can not be considered representative of Italian gynecologists, our data suggest the need of an intensive training to increase gynecologists' knowledge about HIV, other blood-borne pathogens and the risk of occupational infections also in order to modify attitudes and practices. PMID- 7498356 TI - [The request for consent in clinical research: a randomized study in healthy subjects]. AB - To compare two strategies of consent requirement (classical informed consent and randomised consent according to Zelen), the Clinical Data Elaboration Centre of South Italy, within the special project Clinical Application of Oncological Research of the National Research Council of Italy (CNR-ACRO) invited healthy people visiting the 7th edition of the scientific exhibition "Futuro Remoto" to simulate of being ill and receiving the offer of entering a clinical trial. Within informed consent strategy patients are asked to agree to the randomisation process, while, in the randomised consent, randomised treatment assignment is performed before consent requirement and patients should agree directly to the assigned therapy. Major aims of the study were (a) to compare the strategies in terms of refusal rate to a hypothetical clinical trial, and (b) to estimate whether severity of prognosis affected subjects' decision. 3,217 visiting people participated to the simulation; they were prevalently young, males and with a high level of school education. The study was performed in two different scenarios. In the first one, with one choice option, subject refusing consent could receive standard therapy only; refusal rate was 16% after informed consent and 13.4% after randomised consent (for experimental therapy). In the other scenario, with two choice options, subjects refusing consent could choose the preferred therapy; refusal rate was 20.6% after informed consent, 48.1% after randomised consent (for standard therapy) and 13.4% after randomised consent (for experimental therapy).(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7498357 TI - [An interview with Paul Kleihues. Interview by Giuseppe Lojacono]. PMID- 7498358 TI - Disconnected. PMID- 7498359 TI - Anti-CD3-activated splenocytes enhance survival in lethally irradiated mice after transplant of syngeneic hematopoietic stem cells. AB - Although cytokines produced by activated T cells may accelerate immunohematopoietic reconstitution after autologous bone marrow transplantation (ABMT), there is no direct evidence that infusion of anti-CD3 mAb-activated T cells can accelerate engraftment by hematopoietic stem cells. This study tests the ability of anti-CD3-activated murine splenocytes (ASC) to enhance the rescue of lethally irradiated (9 Gy) BDF1 mice by transplant of a limiting dose of fresh unmanipulated syngeneic splenocytes (SC). A minority (14.8%, 10-25%) of mice could be rescued with 5 x 10(5) SC after 9 Gy total-body irradiation (TBI). When 10(6) or 10(7) ASC were added to 5 x 10(5) SC, survival increased to 50% in those that received 5 x 10(5) SC + 10(6) ASC (not significant [NS]) and to 81.4% (77.7 88.0%) in those that received 5 x 10(5) SC + 10(7) ASC (p < 0.001). Furthermore, adding a fixed dose of 10(7) ASC to increasing doses of SC (10(5), 5 x 10(5), and 10(6)) enhanced survival at the different doses of SC. ASC alone did not rescue mice. CD3+ cells were the predominant population (77.6 +/- 6.7%) in the ASC inoculum, while NK cells remained low (1.2 +/- 0.9%). Colony-forming unit-spleen (CFU-S) yield after injection of SC showed dose dependence, whereas injection of 10 x 10(6) ASC alone failed to show any CFU-S yield in 23 of 25 recipient spleens. These results show that ASC enhanced survival of mice rescued with limiting doses of SC and that this effect was ASC dose-dependent but not dependent on the addition of extra stem cells. PMID- 7498360 TI - Circulating CFU-E during hematopoietic recovery after allogeneic bone marrow transplantation: relationship to erythroid engraftment. AB - The more mature erythroid progenitor assayable in vitro, the colony-forming unit erythroid (CFU-E), is normally found in the bone marrow (BM) but is virtually absent from peripheral blood (PB), unlike the more immature progenitor, the burst forming unit-erythroid (BFU-E). We report on the detection of CFU-E in the PB of six of 18 patients during hematopoietic recovery following allogeneic bone marrow transplantation (BMT); three of six patients with PB CFU-E were under treatment with recombinant human erythropoietin (rhEpo) as well as six of 12 who did not present with PB CFU-E. PB CFU-E were found as early as day 14 following BMT, reached a peak on day 28, and were still detectable on day 60. The presence of PB CFU-E was associated with signs of stimulated erythroid engraftment--an accelerated reticulocyte recovery, an increased number of reticulocytes, higher levels of serum transferrin receptor, and a reduction in transfusional requirements were found in these patients compared to those without PB CFU-E. The numbers of PB and BM BFU-E were similar in the two groups, as well as the numbers of PB and BM CFU-granulocyte/macrophage (CFU-GM) and multipotential CFU (CFU GEMM); on the other hand, the percentage of BM BFU-E in S phase of the cell cycle was higher in the group of patients with PB CFU-E. While there was no difference between the two groups in serum Epo levels assayed on days 14 and 28 after BMT, patients with PB CFU-E had higher Epo levels in serum samples collected before starting the BMT procedure. These data suggest that the appearance of circulating CFU-E early after BMT is characteristic of a group of patients with an accelerated erythroid engraftment, although the mechanisms leading to the circulation of CFU-E after BMT remain unclear. PMID- 7498361 TI - Localization of an essential ligand binding determinant of the human erythropoietin receptor to a domain N-terminal to the WSXWS motif: implications for soluble receptor function. AB - The interaction of erythropoietin (Epo) with the erythropoietin receptor (EpoR) supports erythropoiesis. The EpoR is a member of the well-recognized cytokine receptor superfamily characterized by four conserved cysteines and a WSXWS domain in the extracellular portion of the molecule. To localize ligand-binding determinants of the EpoR near the WSXWS domain, we tested the ligand-binding ability of the wild-type human EpoR extracellular domain (EREx), two truncated and three chimeric constructs with the interleukin-2 receptor beta subunit (IL2R beta). Constructs were expressed in E. coli as GST fusion proteins linked to a solid-phase support and assayed for binding to 125I Epo. As previously shown, Epo bound specifically to the expressed extracellular domain, EREx. Epo did not bind to truncated receptors lacking either the entire fifth exon or the WSXWS domain. Epo also did not bind to chimeric receptors that had the amino acids encoded by the fifth exon replaced by IL2R beta or that had the amino acids subsequent to asparagine residue 209 replaced by IL2R beta. Specific binding was demonstrated for a construct in which the WSXWS was replaced by that of IL2R beta. We conclude that the amino acids encoded by this 5' portion of exon 5 of the EpoR are necessary for ligand binding and that the WSXWS domain is necessary for Epo binding but is not involved in ligand-binding specificity. We also speculate that if the putative soluble form of the EpoR is expressed (predicted to lack exon 5), it does not bind Epo and therefore may serve a physiologic purpose other than ligand binding. PMID- 7498362 TI - The synthesis of a peanut agglutinin-ricin A chain conjugate: potential as an in vitro purging agent for autologous bone marrow in multiple myeloma. AB - The lectin peanut agglutinin (PNA) and CD19 monoclonal antibody have been covalently linked to magnetic beads and utilized in an in vitro purging system for autologous bone marrow in multiple myeloma (MM). An alternative to mechanical purging involves the use of immunotoxins to provide specifically targeted cellular toxicity; however, no studies to date have examined the utility of a lectin-ricin A chain (RCA) combination as a purging agent in MM. Initially, we studied the internalization of PNA by target cells (Raji) using flow cytometry. The surface fluorescence intensity of PNA-treated Raji cells was reduced upon incubation at 37 degrees C, and subsequent studies with fixed cells detected the endocytosed PNA. Complete internalization occurred within 120 minutes, indicating the potential of PNA as a purging agent. We manufactured a novel PNA-RCA conjugate and demonstrated its strong and specific binding to PNA reactive cell targets. Subsequent experiments assessed the toxicity of the conjugate to Raji cells and normal bone marrow progenitor cells. 3H-leucine uptake assays showed that PNA-RCA was capable of reducing protein synthesis in Raji cells and that the toxic effects were specific. In addition, at concentrations of conjugate achieving greater than 99% selective cytotoxicity for Raji cells, adequate CFU-GM were preserved in normal marrow. These studies suggest that PNA-RCA may be of value as an in vitro purging agent for MM. PMID- 7498363 TI - Inhibition of leukemic cell proliferation by one or more factors released from splenic BCL1 cells. AB - Irradiated peripheral blood lymphocytes from patients with chronic B-lymphocytic leukemia (B-CLL) have been shown to secrete a factor or factors that caused inhibition of malignant cell proliferation. In the present study, we used the murine B cell leukemia (BCL1) model system to examine the possible secretion of inhibitory factors from irradiated leukemic spleen cells. It was found that under culture conditions, irradiated spleen cells obtained from leukemic mice produce factors capable of suppressing BCL1 cell proliferation in vitro. The release of an inhibitory factor from nonirradiated cells was also observed, albeit to a lower extent. Supernatants collected from cultured nonirradiated and irradiated cells derived from animals at the first week after BCL1 cell inoculation did not affect the proliferation capacity of cells at any stage of the disease, whereas supernatants obtained at more advanced stages exhibited an inhibitory effect on the proliferation of cells derived from the same stages. The suppressive effect of supernatants of irradiated cells was more pronounced when those of 7 days' culture were used, compared to those of nonirradiated cells. The difference in suppression observed between supernatants of irradiated and nonirradiated BCL1 cells may be attributed either to the same factor produced in a larger amount following irradiation or to two (or more) factors--one produced spontaneously, the other induced by irradiation. PMID- 7498364 TI - Effect of the immunomodulator AS101 on chemotherapy-induced multilineage myelosuppression, thrombocytopenia, and anemia in mice. AB - The immunomodulator AS101 has previously been found to induce mouse and human hematopoietic cells to secrete cytokines such as interleukin-1 alpha (IL-1 alpha), IL-2, tumor necrosis factor-alpha (TNF-alpha), and gamma interferon (IFN gamma). The compound was shown to protect mice from lethal and sublethal effects of chemotherapy and irradiation. AS101 prevented the decrease in the number of bone marrow (BM) and spleen myeloid progenitor cells, and increased the survival of lethally treated mice. In this study, we show a dose-dependent response of AS101 in the induction of high secretion levels of IL-6, IL-3, granulocyte macrophage colony-stimulating factor (GM-CSF), and stem cell factor (SCF). Since these growth factors are known to induce the proliferation and differentiation of multilineage progenitors, including megakaryocytic and erythroid progenitors, we designed this study to evaluate the role of AS101 in attenuating thrombocytopenia, anemia, and multilineage myelosuppression associated with chemotherapy. We demonstrate that pretreatment of mice with AS101 24 hours before intraperitoneal injection of 250 mg/kg cyclophosphamide (CYP) or intravenous injection of 150 mg/kg 5-fluorouracil (5-FU) significantly increased the number of circulating white blood cells (WBC) and platelets. The numbers of both neutrophils and lymphocytes were significantly increased in AS101-treated mice subjected to chemotherapy. In addition, AS101 attenuated erythropenia caused by 5 FU. It could also increase megakaryocyte and erythroid progenitor cells (CFU-MK and CFU-E) in the BM of treated mice severely affected by chemotherapy. We demonstrate that the protective effect of AS101 could be abrogated by treatment with anti-IL-1R or anti-SCF antibodies. We suggest that the endogenous production of cytokines such as IL-1, IL-6, IL-3, SCF, and GM-CSF in mice treated with AS101 offers protection to circulating blood elements and ameliorates the reconstitution of megakaryocytic and erythroid progenitors. The simultaneous protection by AS101 of multilineage cell compartments is probably due to stimulation by AS101 of a selective subpopulation of primitive stem cells resistant to chemotherapy. On the basis of these studies, phase II clinical trials with patients treated with chemotherapy in combination with AS101 have been initiated. PMID- 7498365 TI - Azidothymidine and interferon-alpha in vitro effects on hematopoiesis: protective in vitro activity of IL-1 and GM-CSF. AB - Preclinical and clinical studies with an azidothymidine (AZT)/interferon-alpha (IFN-alpha) combination resulted in a marked and synergistic antiretroviral activity. The administration of the two drugs in HIV-seropositive patients affected with Kaposi's sarcoma, however, induced neutropenia, thrombocytopenia, and, in some cases, anemia. A possible means to improve the therapeutic index of AZT and/or IFN-alpha in AIDS patients could be the addition of hematopoietic growth factors. In vitro activity of cytokines on the hematotoxicity of the AZT IFN-alpha association has not yet been studied. We have performed an in vitro study to evaluate the toxicity of AZT, IFN-alpha, or both on peripheral blood hematopoietic progenitors (CFU-GM and BFU-E) and to assess the activity of interleukin 1 (IL-1), granulocyte-macrophage colony-stimulating factor (GM-CSF), or both in modifying AZT-IFN-alpha hematotoxicity. Results indicate that AZT, IFN alpha, and combinations of the two have a dose-dependent inhibitory effect on the in vitro growth of peripheral blood hematopoietic progenitors. Combinations of AZT and IFN-alpha inhibited CFU-GM and BFU-E proliferation in an additive manner. Neither IL-1 nor GM-CSF alone was able to induce a significant reduction of AZT induced damage. Only the addition to the cultures of both cytokines partially curbed the antiproliferative activity of AZT at low dosages. PMID- 7498366 TI - Effects of low oxygen tension and antioxidants on human erythropoiesis in vitro. AB - Cultured cells grow better in low oxygen than in room air, and the addition of a sulfhydryl-reducing agent is usually routine. It has been thought that such reagents protect cells from free-radical toxicity in atmospheric oxygen, which is higher than in physiological oxygen tension. Previous data demonstrating the efficacy of the usual antioxidant, 2-mercaptoethanol (2ME), were obtained in murine studies. We have compared the effect of 2ME and other antioxidants on human erythropoiesis using neonatal (cord) and adult blood. Without antioxidants, erythroid colony numbers were higher in 5% oxygen than in air, while colony formation could be improved even in 5% oxygen by the addition of antioxidants. In dose-response curves of each drug, the maximum increase in colony number was achieved with 0.1 microM 2ME, 10 microM reduced glutathione (GSH), 1 U/mL catalase (CAT), and 1 U/mL superoxide dismutase (SOD). Erythroid colony growth in air and in 5% oxygen was identical with 10 microM GSH and higher than in any other conditions, including with combinations of GSH and any other antioxidant. For human erythroid progenitor cell cultures, 10 microM GSH appears to be more appropriate than the usual 100 microM 2ME. Antioxidants are necessary, however, even in low oxygen tension. The mechanism of the GSH action is incompletely understood. PMID- 7498368 TI - Production of thrombopoietin (TPO) by rat hepatocytes and hepatoma cell lines. AB - Recently, we purified rat thrombopoietin (TPO) from plasma of irradiated rats (XRP) by measuring its activity that stimulated the production of megakaryocytes from megakaryocyte progenitor cells (CFU-MK) in vitro. We then cloned the cDNAs for rat and human TPO. In this study, we found the production of TPO by hepatocytes isolated with the collagenase perfusion method from both normal and thrombocytopenic rats, by a two-step fractionation of hepatocyte culture medium (CM). Subsequently, CM of rat hepatoma cell lines was screened for the presence of TPO; three cell lines, H4-II-E, McA-RH8994, and HTC, were found to produce TPO. According to the purification procedure for TPO from XRP, TPO was partially purified from 2 L CM of each of three cell lines with a six-step procedure. In the final reverse-phase column, TPO from each cell line was eluted with the same retention time as that from XRP, and the TPO fraction exhibited megakaryocyte colony-stimulating activity (Meg-CSA). TPO-active fraction eluted from the final reverse-phase column was separated by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE), extracted from the gel, and assayed. TPO activity from each cell line was found in the respective molecular weight region, indicating the heterogeneity of the TPO molecule. Using reverse transcriptase polymerase chain reaction (RT-PCR), we detected the expression of TPO mRNA in hepatocytes, three hepatoma cell lines, normal rat liver, and X-irradiated rat liver. Northern blot analysis showed that TPO mRNA was expressed mainly in liver among the various organs tested. These data demonstrate that TPO is produced by rat hepatocytes and hepatoma cell lines and suggest that liver may be the primary organ that produces TPO. PMID- 7498367 TI - Effects of bone marrow fibroblasts on the proliferation and differentiation of myeloid leukemic cell lines. AB - The effects of normal bone marrow fibroblasts (BM FB) on proliferation and differentiation of 10 myeloid leukemic cell lines were investigated in a serum free co-culture system. The proliferation of three of the cell lines was supported by BM FB. Three of the myeloid cell lines were inhibited 40-70%. The co culture supernatants were tested for the secretion of hematopoietic cytokines by bioassays. Except for IL-6, which was already produced constitutively by BM FB, only little amounts of interleukin-1 (IL-1), granulocyte colony-stimulating factor (G-CSF), or granulocyte-macrophage colony-stimulating factor (GM-CSF) could be detected in several co-culture supernatants. It could be shown that, according to cytologic and functional criteria, the myeloid leukemic cell lines ML-2 and PLB-985 differentiate along the monocyte-macrophage pathway after co culture with BM FB. They revealed a histiocytic phenotype and could be induced to produce reactive oxygen intermediates (ROI) after stimulation with zymosan or phorbol-myristate-acetate (PMA). Additional proof for differentiation was obtained from flow cytometric analysis of surface differentiation antigens and adhesion molecules. The neutralization of IL-6 activity in the co-cultures by antibodies resulted in prevention of differentiation of PLB-985 cells, while differentiation of ML-2 cells in the co-cultures was not affected by addition of anti-IL-6 antibodies. Furthermore, in co-culture experiments with fibroblasts from skin and foreskin, we found a differentiation of PLB-985 cells comparable to that in co-cultures with BM FB, but poor differentiation of ML-2 cells. These data suggest that different mechanisms are involved in the differentiation of ML 2 and PLB-985 cells. PMID- 7498370 TI - Isolation of endothelial-like stromal cells that express Kit ligand and support in vitro hematopoiesis. AB - Although macrophages account for 70-90% of the adherent cells in mouse long-term bone marrow cultures (LTBMC) and CFU-F colonies, the predominant nonhematopoietic stromal cell is endothelial-like (EL), expressing cytoplasmic collagen IV, laminin, and an antigen recognized by the monoclonal antibody MECA-10. We report the isolation of this stromal cell lineage from primary LTBMC by immunomagnetic cell selection using MECA-10. More than 95% of the cells in the MECA-10-positive fraction are EL cells as judged by morphology, surface staining for MECA-10, cytoplasmic staining for collagen IV, and electrophoretic analysis of MECA-10 positive cells isolated from radiation chimeras. When plated under LTBMC conditions, EL cell monolayers recharged with either wild-type or Sl/Sld marrow support an increased density and number of clonogenic and mature hematopoietic cells in short-term cultures. In accord with this finding, Northern blots of mRNA from unstimulated EL cells demonstrate constitutive expression of Kit ligand (KL). Moreover, in situ two-color immunofluorescence staining for cytoplasmic collagen IV and surface KL suggests that EL cells are the exclusive source of membrane-bound KL in mouse cultures. The ability to isolate EL cells from primary cultures without the need for repeated cell passage or immortalization provides a novel approach to dissecting the molecular basis of stem cell-stromal cell interactions. PMID- 7498369 TI - Transgenic mice expressing either bovine growth hormone (bGH) or human GH releasing hormone (hGRH) have increased splenic progenitor cell colony formation and DNA synthesis in vitro and in vivo. AB - To investigate the potential effects of growth hormone (GH) on the hematopoietic system, mice transgenic for bovine GH (bGH) or human growth hormone releasing hormone (hGRH) genes, each of which can result in the constitutive overproduction of GH, were analyzed for splenic and bone marrow (BM) localized hematopoietic progenitor cells. These transgenic mice had splenic hyperplasia with increased absolute numbers of splenic erythroid and megakaryocytic progenitor cells as assessed by in vitro assay and megakaryocyte development as seen in spleens. As an in vivo indication of multilineage progenitor cell effects in hGRH mice, the number of day-10 CFU-S colonies derived from the donor spleen was significantly higher than in nontransgenic littermate controls. A high proportion (54-71%) of splenic erythroid, granulocyte-macrophage, and megakaryocyte progenitors were in cycle in transgenic mice in contrast to < or = 30% in nontransgenic control littermates. Compared to controls, splenocytes from hGRH mice had a significantly higher proliferative index when infused into irradiated nontransgenic controls. With the exception of the megakaryocyte colony assay and in vivo proliferative index, none of these findings were evident when identical assays were performed on the BM from the same mice. Consistent with the BM data, peripheral blood leukocyte, erythroid, and platelet numbers were comparable in transgenic and nontransgenic control littermates. We conclude that the constitutive expression of bGH or hGRH leads largely to splenic hematopoietic effects involving progenitor cell populations from at least two lineages. PMID- 7498371 TI - Monosynaptic rubrospinal projections to distal forelimb motoneurons in the cat. AB - In the present electrophysiological experiments, we examined whether or not there are short-latency excitatory postsynaptic potentials (EPSPs) suggesting monosynaptic rubro- and corticomotoneuronal connections in the cervical spinal segments in cats. Under pentobarbital anesthesia and immobilization with gallamine triethiodide, intracellular recordings were made from forelimb motoneurons (Mns) following stimulation of the red nucleus (RN) and the cerebral peduncle (CP). Mns comprising the musculocutaneous nerve (Musc. Mns), the radial nerve (Rad. Mns), the ulnar nerve (Uln. Mns), and the median nerve (Med. Mns) were identified by their antidromic activation. Single-pulse stimulation of RN produced EPSPs with a mean amplitude of 0.35 mV in the majority of forelimb Mns (88 of 122, 72%). Half of the forelimb Mns (23 of 46) of the C8-T1 segments in which RN-EPSPs were detected by single shocks exhibited segmental latencies of RN EPSPs shorter than 1.0 ms, while only 3 of 43 (7%) forelimb Mns in the C6-7 segments showed such latencies. These results suggest that rubromotoneuronal connections are, at least in part, monosynaptic for forelimb Mns that innervate in particular the hand and digit muscles. When paired-pulse stimuli (interpulse interval of 3 ms) were applied to RN, the second EPSPs were potentiated moderately (mean amount of temporal facilitation 24%) for EPSPs with short segmental latencies (< 1.0 ms). By contrast, single shocks to CP rarely produced EPSPs in forelimb Mns (9 of 106, 8%). There was no obvious evidence to suggest monosynaptic connections between corticospinal fibers and the forelimb Mns. The present results suggest the presence of preferential direct rubrospinal projection to Mns innervating distal forelimb muscles in the cat. PMID- 7498373 TI - Systematic differences in time of dopaminergic neuron origin between normal mice and homozygous weaver mutants. AB - Immunocytochemical labeling for tyrosine hydroxylase and [3H]thymidine autoradiography were combined in wild-type mice and in mice homozygous for the weaver mutant gene (wv) to see whether the neurogenetic patterns of midbrain dopaminergic neurons was normal in the mutants and whether the degeneration of dopaminergic neurons was linked to their time of origin. Dams of wild-type and homozygous weaver mice were injected with [3H]thymidine on embryonic days (E) 11 E12, E12-E13, E13-E14, and E14-E15 to label neurons in the retrorubral field, the substantia nigra pars compacta, the ventral tegmental area, and the interfascicular nucleus as they were being generated. The quantitatively determined time of origin profiles indicated that wv/wv mice have the same time span of neurogenesis as +/+ mice (E10 to E14), but have significant deficits in the proportion of late-generated neurons in each dopaminergic population. In the retrorubral field and substantia nigra, weaver homozygotes had substantial losses of dopaminergic neurons and had a greater deficit in the proportion of neurons generated late while, in the ventral tegmental area and interfascicular nucleus, there were slight losses of dopaminergic neurons and only slight deficits in the proportion of late-generated neurons. These findings lead to the conclusion that the weaver gene is specifically targeting dopaminergic neurons that are generated late, mainly on E13 and E14. PMID- 7498372 TI - Time of neuron origin and gradients of neurogenesis in midbrain dopaminergic neurons in the mouse. AB - Previous [3H]thymidine studies in Nissl-stained sections in rats established that the substantia nigra pars compacta and the ventral tegmental area originate sequentially according to an anterolateral to posteromedial neurogenetic gradient. We investigated whether that same pattern is found in mice in the dopaminergic neurons in each of these structures. Using tyrosine hydroxylase immunostaining combined with [3H]thymidine autoradiography, the time of origin of dopaminergic midbrain neurons in the retrorubral field, the substantia nigra pars compacta, the ventral tegmental area, and the interfascicular nucleus was determined in postnatal day 20 mice. The dams of the experimental animals were injected with [3H]thymidine on embryonic days (E) 11-E12, E12-E13, E13-E14, and E14-E15. The time of origin profiles for each group indicated significant differences between populations. The retrorubral field and the substantia nigra pars compacta arose nearly simultaneously and contained the highest proportion of neurons, 49 to 37%, generated on or before E11. Progressively fewer early generated neurons were found in the ventral tegmental area (20%), and the interfascicular nucleus (8.5%). In addition, anterior dorsolateral neurons in the substantia nigra and ventral tegmental area were more likely to be generated early than the posterior ventromedial neurons. These findings indicate that mouse and rat brains have nearly identical developmental patterns in the midbrain, and neurogenetic gradients in dopaminergic neurons are similar to those found in Nissl studies in rats. PMID- 7498374 TI - Interactions between two different visual stimuli in the receptive fields of inferior temporal neurons in macaques during matching behaviors. AB - Macaque monkeys were trained to determine whether shapes or colors of two visual stimuli were the same or different (matched/non-matched). Two stimuli were presented at different locations while the monkey fixated a small spot. In one paradigm, two timuli were presented simultaneously for 0.5 s (Sml-SO task). In the other paradigm, one of the stimuli was turned on 0.5 s before the onset of another stimulus, then the two stimuli were present for the following 0.5 s (Scc SO task). The aim of the later task was to analyze the responses of TE neurons to a single presentation of each stimulus and the effects of successive onsets of two stimuli. Of 232 responsive neurons tested in both tasks, 143 showed a significant selectivity between paired stimuli (termed 'selective neurons'). During the Sml-SO task, some selective neurons showed a larger response to the different (non-matched) stimuli than to the double optimal stimuli (matched), even though one of the different stimuli was inhibitory. This effect was more prominent in neurons that showed a smaller response to the double presentations of the optimal stimulus than to the single presentation. Since another group of selective neurons showed smaller responses for the different stimuli, the average response amplitudes were similar between the identical and the different stimuli. During the Scc-SO task, when the optimal stimulus was turned on after the non optimal stimulus (non-matched), the response to the second stimulus was mostly enhanced above the response level to the single presentation of the first stimulus. Since the responses to the second stimulus identical to the first stimulus tended to decrease, the difference in the responses between the matched and non-matched stimuli became significantly larger in the Scc-SO task. The reaction times of the monkeys were shorter during the Scc-SO task than the Sml-SO task. These changes in response amplitude between the different and the identical stimuli were not prominent in the non-selective neurons. These results suggest that non-linear interactions between two different stimuli play an important role in the discrimination of groups of visual stimuli, particularly in successive analysis. PMID- 7498376 TI - Neural responses to free-field auditory stimulation in the superior colliculus of the wallaby (Macropus eugenii). AB - Auditory responses to free-field broad band stimulation from different directions were recorded from clusters of neurones in the superior colliculus (SC) of the anaesthetized tammar wallaby. The auditory responses were found approximately 2 mm beneath the first recording of visually evoked responses in the superficial layers, the vast majority being solely auditory in nature; only one recording responded to both auditory and visual stimulation. Responses to suprathreshold intensities displayed sharp spatial tuning to sound in the contralateral hemifield. Those from the rostral pole of the SC disclosed a preference for auditory stimuli in the azimuthal anterior field, whereas those in the caudal SC preferentially responded to sounds in the posterior field. A continuum of directionally tuned responses was seen along the rostrocaudal axis of the SC so that the entire azimuthal contralateral auditory hemifield was represented in the SC. Furthermore, tight spatial alignment was evident between the best position of the visual responses in the superficial layers in azimuth and the peak angle of the auditory response in the deeper layers. PMID- 7498375 TI - Cyclophosphamide cystitis as a model of visceral pain in rats: model elaboration and spinal structures involved as revealed by the expression of c-Fos and Krox-24 proteins. AB - The evoked expression of the immediate early gene (IEG)-encoded proteins c-Fos and Krox-24 was used to monitor spinal visceronociceptive processing that results from cyclophosphamide cystitis in behaving rats. Animals received a single dose of 100 mg/kg i.p. of cyclophosphamide and survived for 30 min to 5 h. Longer survival times were not considered because of ethical considerations. Cyclophosphamide-injected animals developed characteristic behavioral signs in parallel with development of bladder lesions and spinal evoked expression of IEG encoded proteins. Histological examination of the urinary bladder was used to evaluate the degree of cystitis and as a criterion for selection of groups of animals to be quantitatively analyzed. Controls consisted of freely behaving animals including control (un-injected), sham (saline-injected) or diuretic (furosemide-injected) animals. Behavioral modifications consisted of lacrimation, piloerection, assumption of a peculiar "rounded-back" posture, which was accompanied by head immobility and various brief "crises" (tail hyperextension, abdominal retractions, licking of the lower abdomen, backward withdrawal movements). Abnormal behaviors, which first appeared (lacrimation, piloerection) at the end of postinjection hour 1, progressively increased in severity (rounded back posture) over the following 90 min to reach a plateau at about postinjection hour 2; the rounded-back posture was maintained up to time of death. Histological modifications of bladder tissue were assessed using a 4-grade scale in a blind setting. The 1st grade consisted of control or sham animals with no bladder lesion; 2nd grade, animals with simple chorionic edema; 3rd grade, animals with chorionic edema associated with mucosal abrasion, fibrin deposit, and onset of polymorphonuclear leukocyte infiltration; 4th grade, animals with complete cystitis corresponding to an increase in severity and spread of all the signs of cystitis described above plus petechial hemorrhage. Simple chorionic edema was observed from 30 min to 3 h postinjection, but with a progressive increase in severity over time. Edema accompanied by epithelial abrasion was observed for animals that survived 3-4 h postinjection; complete inflammation was observed in animals that survived 4-5 h postinjection. The study of c-Fos- and Krox-24 encoded protein expression demonstrated that few lumbosacral spinal areas were specifically involved in the processing of visceral inputs in response to bladder stimulation. These areas were the parasympathetic column (SPN), the dorsal gray commissure (DGC as the caudal extent of lamina X), and superficial layers of the dorsal horn.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 7498377 TI - Distribution of aspartate and glutamate in the nucleus of the solitary tract of the lamb. AB - Excitatory amino acids have been implicated in several nucleus of the solitary tract (NST)-mediated functions. The distribution of the excitatory amino acids aspartate and glutamate has been described in both cat and rat. However, the distribution of these amino acids has not been described for the lamb, a species frequently used in the investigation of NST-mediated behaviors. Thus, this study was designed to investigate the distribution of aspartate-like (ASP) and glutamate-like (GLU) immunoreactivity in the lamb NST using pre- and post embedding immunohistochemistry. Both ASP- and GLU-immunoreactive cells and puncta were observed throughout the rostral to caudal extent of the lamb NST. The most intense ASP- and GLU-immunoreactive cell and puncta staining was found ventromedial, ventral and ventrolateral to the solitary tract at intermediate and caudal levels of the lamb NST. The relative numbers of both cells and puncta stained were lower at rostral levels of the NST corresponding to the gustatory NST. The intense ASP and GLU immunoreactivity observed in areas of the lamb intermediate and caudal NST that are involved in respiration, deglutition and cardiovascular functions suggests excitatory amino acids plays an important role in NST neural processing that underlies these behaviors in lamb. PMID- 7498378 TI - Visual tuning to kinematics of biological motion: the role of eye movements. AB - The visual system is particularly sensitive to the covariation between velocity and curvature that constraint biological motion. Previous work showed that, when this biological constraint is satisfied, simple elliptical motion of a dot looks constant, although its velocity is highly non-uniform. This paper addresses the hypothesis that such a dynamic illusion is dependent upon smooth pursuit eye movements. Subjects had to adjust the kinematics of a dot moving along elliptical trajectories until they perceived a constant velocity. Different pursuit and fixation conditions were tested. The research shows that the dynamic illusion is largely independent of eye movements, suggesting that the visual system has access to implicit knowledge of motor constraints regardless of the concurrent oculomotor commands. PMID- 7498379 TI - Spatial attention and eye movements. AB - We previously showed that when attention is allocated to the right or left of the fixation point, saccades directed to targets located above or below the fixation point deviate contralateral to the attention locus. In the present study, we examined how general this phenomenon is and whether the amount of saccade deviation depends on the location of attention with respect to that of the saccade target. Three experiments were carried out. In experiment 1 the location of the imperative stimulus was uncued. Its presentation exogenously directed attention to its location. In experiment 2 the location of the imperative stimulus was cued by a central cognitive cue. In this experiment attention was endogenously directed to the imperative stimulus location before its presentation (expectancy paradigm). In experiment 3 all stimulus boxes contained a possible imperative stimulus at the display presentation. A central cue, presented subsequently, indicated which of them had to be used for the saccade. In this experiment attention was endogenously directed to the imperative stimulus, but after its presentation (no-expectancy paradigm). The results showed that, regardless of how attention was directed to the imperative stimulus, the vertical saccades deviated contralateral to the attention location. The deviation was larger when attention was in the upper field and the saccade was directed upward ("same hemifield" condition) than when attention was in the upper field and the saccade was directed downward ("opposite hemifield" condition). The same relationship between the "same hemifield" condition and "opposite hemifield" condition was found when attention was in the lower field. Saccadic reaction times (SRTs) were shortest in experiment 2 and longest in experiment 3. In experiment 2, SRTs of the "same hemifield" condition were significantly longer than those of the "opposite hemifield" condition. Taken altogether, these results strongly support the notion that attention allocation in space leads to an activation of oculomotor circuits, in spite of eye immobility. The possible mechanisms responsible for saccade deviations and for greater saccade deviations when attention is in the same hemifield as the programmed ocular saccade are discussed. PMID- 7498380 TI - Focal depression of cortical excitability induced by fatiguing muscle contraction: a transcranial magnetic stimulation study. AB - Motor evoked potentials (MEPs) elicited by transcranial magnetic stimulation (TMS) and transcranial electrical stimulation (TES) of the motor cortex were recorded in separate sessions to assess changes in motor cortex excitability after a fatiguing isometric maximal voluntary contraction (MVC) of the right ankle dorsal flexor muscles. Five healthy male subjects, aged 37.4 +/- 4.2 years (mean +/- SE), were seated in a chair equipped with a load cell to measure dorsiflexion force. TMS or TES was delivered over the scalp vertex before and after a fatiguing MVC, which was maintained until force decreased by 50%. MEPs were recorded by surface electrodes placed over quadriceps, hamstrings, tibialis anterior (TA), and soleus muscles bilaterally. M-waves were elicited from the exercised TA by supramaximal electrical stimulation of the peroneal nerve. H reflex and MVC recovery after fatiguing, sustained MVC were also studied independently in additional sessions. TMS-induced MEPs were significantly reduced for 20 min following MVC, but only in the exercised TA muscle. Comparing TMS and TES mean MEP amplitudes, we found that, over the first 5 min following the fatiguing MVC, they were decreased by about 55% for each. M-wave responses were unchanged. H-reflex amplitude and MVC force recovered within the 1st min following the fatiguing MVC. When neuromuscular fatigue was induced by tetanic motor point stimulation of the TA, TMS-induced MEP amplitudes remained unchanged. These findings suggest that the observed decrease in MEP amplitude represents a focal reduction of cortical excitability following a fatiguing motor task and may be caused by intracortical and/or subcortical inhibitory mechanisms. PMID- 7498381 TI - Influence of gastrocnemius muscle length on triceps surae torque development and electromyographic activity in man. AB - The present study was designed to determine the relative contribution of the gastrocnemius muscle to isometric plantar flexor torque production at varying knee angles, while investigating the activation of the gastrocnemius muscle at standardised non-optimal lengths. Voluntary plantar flexor torque, supramaximally stimulated twitch torque and myoelectric activity (EMG) from the triceps surae were measured at different knee angles. Surface and intra-muscular EMG were recorded from the soleus muscle and the medial and lateral heads of the gastrocnemius muscle in 10 male subjects. With the ankle angle held constant, knee angle was changed in steps of 30 degrees ranging from 180 degrees (extended) to 60 degrees (extreme flexion), while voluntary torque from a 5-s contraction was determined at 10 different levels of voluntary effort, ranging from 10% of maximal effort to maximal effort. To assess effort, supramaximal twitches were superimposed on all voluntary contractions, and additionally during rest. Maximal plantar flexor torque and resting twitch torque decreased significantly in a sigmoidal fashion with increasing knee flexion to 60% of the maximum torque at 180 degrees knee angle. For similar levels of voluntary effort, the EMG root mean square (RMS) of gastrocnemius was less with increased knee flexion, whereas soleus RMS remained unchanged. From these data, it is concluded that the contribution of gastrocnemius to plantar flexor torque is at least 40% of the total torque in the straight leg position. The decrease of gastrocnemius EMG RMS with decreasing muscle length may be brought about by a decrease in the number of fibres within the EMG electrode recording volume and/or impaired neuromuscular transmission. PMID- 7498382 TI - Unconscious updating of grasp motor program. AB - Grasp modification during prehension movements was studied in response to slight variations of somesthetic information about object size. Three experiments were carried out. In experiment 1 eight subjects were required to reach and grasp an object whose size could either increase or decrease, whereas its visual image remained unmodified. The object size was changed during the experiment with uninformed subjects after a block of trials during which visual and somesthetic information were congruent. At the end of the experiment subjects were required to reproduce the size of the object with their fingers (matching test). Results showed that maximal grip aperture during prehension as well as finger aperture in the matching test were modified according to variation in object size, although no subject realized that the object had changed during the experiment. Grasp time was also altered by object size change. Greater and earlier adaptation in maximal grip aperture, as well as perturbation of grasp time, were observed for decrease than for increase in object size. However, complete compensation was never reached for both parameters. Constant confidence in vision could have prevented both complete compensation and conscious detection of object change. This was investigated in two additional experiments. In experiment 2 visual information was made unreliable by informing subjects about variation in grasped object size. This led to greater and earlier modification in maximal grip aperture than in experiment 1. Grasp time was kept almost constant regardless of size variation. In experiment 3 vision of the stimulus was prevented and no information on change in object size was given to subjects. The results of experiment 3 were similar to those of experiment 1, although modification in maximal grip aperture was larger for increase in object size. Correspondingly, grasp time was more affected by increase than by decrease in object size. The results of the three experiments suggest that kinematic parameters usually considered as dependent on object properties, such as maximal grip aperture, were modified in order to compensate perturbation of temporal parameters. This modification induced a "pragmatic" knowledge of object size (as showed by the results of the matching test), although awareness was not reached. PMID- 7498383 TI - Asymmetric control of bilateral isometric finger forces. AB - We examined the ability to match the voluntary isometric finger flexion forces of the dominant and nondominant hand in humans, as well as the influence of unilateral visual feedback during this task. Right- and left-handed subjects were trained to produce a "low" force level (50 +/- 25 g) and a "high" force level (200 +/- 50 g) with the right and left index finger, separately. Following the training session, subjects were instructed to match the isometric forces of both fingers simultaneously within the required range (either low or high) so that they were perceived to be identical. The results showed an asymmetry, whereby greater forces were exerted with the index finger of the dominant hand. The asymmetry was independent of the subjects' maximum finger flexion strength. When unilateral visual feedback represented the force output of the dominant hand, the asymmetry was no longer present. In contrast, when it represented the force output of the nondominant hand, the asymmetry was not compensated. We hypothesize that these findings are the result of anatomical or physiological asymmetries inherent in the motor system controlling the production of force. PMID- 7498384 TI - Contribution of tactile afferent information to the control of isometric finger forces. AB - The ability to match the voluntary isometric force output of the right and left index fingers when the contact surfaces differ in shape was examined. Before the experiment, subjects were trained to produce both a "low" force level (50 +/- 25 g) and a "high" force level (200 +/- 50 g) with the right and left index finger, separately. Following the training session, subjects were instructed to match the forces of both fingers simultaneously within the required range (either low or high) so that the forces were perceived to be identical. One of the index fingers pushed against a conical contact pad, while the other pushed against a flat contact pad. Midway through the experiment, the two contact pads were reversed. Subjects consistently produced less force with the finger pressing against the conical pad. This asymmetry could already be seen during the beginning of the ramp increase in force and continued throughout the trial, independent of the target force levels (low or high). These findings suggest tactile afferent information at the finger-tip is important for determining the voluntary force exerted by the finger. It must be properly integrated with other peripheral information as well as with the central motor command, otherwise the perception of force is distorted. Furthermore, the perception of the force produced seemingly is dependent on the extent to which the skin of the fingertip is indented rather than the local pressure exerted at the skin. PMID- 7498385 TI - Moving one's finger to a visually specified position: target orientation influences the finger's path. AB - It has previously been shown that, when subjects are instructed to move their finger slowly from one point to another the finger follows a path that deviates systematically from a straight line connecting the two points. The deviation depends on the angle between this fictive line and a line connecting the subject's finger with his body. In the present study, we examined whether the deviation also depends on the target's orientation. In two experiments, subjects were instructed to move a finger slowly towards five targets. We recorded the finger's movements. In one experiment, the targets were aligned. In the other, they were oriented radially around the starting point. Otherwise, conditions were the same. The difference in target orientation influenced the finger's path. Most importantly, when the targets were oriented radially around the starting point, the finger's path was straight. We conclude that pointing is more than moving the finger to a specified position. PMID- 7498386 TI - Transcutaneous electrical stimulation and vibration of neck muscles in neglect. AB - Four neglect patients without visual field defects, one with a lesion of the right basal ganglia and three with a right, predominantly parietal lesion, were examined with a cancellation and a copying task before, during and after neck muscle vibration, during transcutaneous electrical stimulation of neck muscles and during vibration of hand muscles on the left side. In all patients, neck muscle vibration improved task performance, while transcutaneous electrical stimulation and hand vibration had little or no effect. The present results demonstrate that the effect of neck muscle vibration cannot be explained as arousal and activation due to unspecific sensory stimulation on the contralesional side of the body. They rather argue for the assumption that the compensatory effect of neck muscle vibration on neglect is an effect induced by the predominant activation of afferent Ia nerve fibres and their specific contribution to the central representation of egocentric space. PMID- 7498387 TI - Relationship of simultaneously recorded cerebellar nuclear neuron discharge to the acquisition of a complex, operantly conditioned forelimb movement in cats. AB - This study was designed to examine the changes in the modulation of small populations of cerebellar neurons during the acquisition of a complex, operantly conditioned forelimb task in cats. The experiments are based on the general postulate that, during the learning of a complex motor behavior, the cerebellum is important for generating a coordinated movement that meets the task's objectives, and that, as the cerebellum participates in this process, it acts to reinforce the effective motor pattern once it has been established. This specific study examines whether the changes in the modulation of cerebellar nuclear neurons during the learning of this task are consistent with this view. Cats were required to learn to move the manipulandum through a novel pattern of 2-3 consecutive straight grooves connected end to end in different spatial configurations, e.g., the letter L, an inverted L, and the letter C. Throughout the acquisition process, 6-12 single units were recorded simultaneously in the cerebellar nuclei, and the kinematics of the movement were evaluated using an Optotrak system. Cells were recorded from the two interposed nuclei and the dentate nucleus in these initial studies. Trials were sorted off-line based on the level of skill at which the required movement was performed. This was assessed using several objective criteria such as movement times, kinematic characteristics, and smoothness (number of peaks in the velocity profile). Event related histograms then were constructed from each group of sorted trials. Changes in modulation related to a specific event were measured in successive histograms for each neuron.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7498389 TI - Spatial coordination by descending vestibular signals. 1. Reflex excitation of neck muscles in alert and decerebrate cats. AB - Electromyographic activity of dorsal neck muscles and neck torques was recorded to study vestibulocollic, cervicocollic, and combined reflexes in alert and decerebrate cats during rotations of the whole body, the body except for the head, and the head but not the rest of the body. Cats were rotated about many axes that lay in the frontal, sagittal, and horizontal planes using sinusoidal 0.25-Hz waveforms or sum-of-sinusoid wave-forms. Robust electromyographic responses were recorded from six muscles, with response directionality that in most cases did not show strong dependence on the reflex tested or on other factors including exact neck angle, stimulus amplitude from 5 degrees to 60 degrees, and intact versus decerebrate state. Based on the strength of responses to rotations about all the tested axes, neck muscles could be characterized by maximal activation direction vectors representing the axis and direction of rotation in three-dimensional space that was most excitatory during reflex responses. Responses to rotations about axes that lay in a coordinate plane were predicted by a cosine function of the angle between the axis under test and the maximally excitatory axis in the plane. All muscles were excited by the nose down phase of pitch rotation and by yaw and roll away from the side on which the muscle lay. Biventer cervicis was best activated by rotations with axes near nose down pitch, and its axis of maximal activation also had small, approximately equal components of yaw and roll toward the contralateral side. Complexus was best excited by rotations with axes nearest roll, but with large components along all three axes. Occipitoscapularis was best excited by rotations about axes near pitch, but with a moderately large contralateral yaw component and a smaller but significant contralateral roll component. Splenius was best excited by rotations with a large component of contralateral yaw, considerable nose-down pitch, and a smaller component of contralateral roll. Rectus major was best excited by rotations near nose-down pitch, but with a substantial contralateral yaw component and smaller contralateral roll component. Obliquus inferior was best excited by rotations with a large component of contralateral yaw, but with considerable contralateral roll and nose-down pitch components. All muscles responded as though they received convergent input from all three semicircular canals. Vestibulocollic and combined reflex responses in alert cats and vestibulocollic, cervicocollic, and combined responses in decerebrate cats appeared to have the same directionality, as evidenced by insignificant shifts in maximal activation vectors.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7498390 TI - The influence of insulin-induced hypoglycemia on the calcium transients accompanying reversible forebrain ischemia in the rat. AB - The primary objective of this study was to explore why preischemic hypoglycemia, which restricts tissue acidosis during the ischemic insult, does not ameliorate cell damage incurred as a result of transient ischemia. The question arose whether hypoglycemia (plasma glucose concentration 2-3 mM) delays resumption of extrusion of Ca2+ from cells during recirculation. Measurements of extracellular Ca2+ concentration during forebrain ischemia of 15 min duration proved that this was the case. Thus, normoglycemic animals resumed Ca2+ extrusion upon recirculation after a delay of 1.5-2.0 min, and hypoglycemic ones after an additional delay which could amount to 3-4 min. We attempted to explore the cause of this delay. At first sight, the results suggested that resumption of oxidative phosphorylation upon recirculation was substrate limited. However, glucose infusion during ischemia or just after recirculation failed to accelerate Ca2+ extrusion from the cells. A comparison between non-injected and insulin-injected animals at equal plasma glucose concentrations suggested that insulin was responsible for the delay. On analysis, the delay proved to be related to a sluggish recovery of cerebral blood flow. The results suggest that when cell damage is evaluated after transient ischemia in hypo- and normoglycemic subjects, attention should be directed to the period of cell calcium 'overload'. Unobserved differences in the duration of the calcium transient may also confound interpretation of data on the effects of insulin. PMID- 7498388 TI - The expression of different cytochemical markers in normal and axotomised dorsal root ganglion cells projecting to the nucleus gracilis in the adult rat. AB - Rat lumbar dorsal root ganglion neurones projecting to the nucleus gracilis in the brainstem were retrogradely labelled with Fluoro-Gold and analysed immunocytochemically for their expression of substance P-, calcitonin gene related peptide-, galanin-, galanin message-associated peptide-, neuropeptide Y-, nitric oxide synthase- and carbonic anhydrase-like immunoreactivity as well as affinity to Griffonia (bandeiraea) simplicifolia lectin I--isolectin B4, RT97 and to choleragenoid. The analysis was made both in uninjured rats and in rats which had been subjected to unilateral sciatic nerve transection and partial resection 3 weeks earlier. The data showed that 6% of the L4 and L5 lumbar dorsal root ganglion cells that projected to the nucleus gracilis showed substance P-like immunoreactivity. Following nerve injury, none of the nucleus gracilis-projecting dorsal root ganglion cells showed substance P-like immunoreactivity. Nineteen per cent of the investigated cell population showed calcitonin gene-related peptide like immunoreactivity in uninjured rats, but no nucleus gracilis-projecting calcitonin gene-related peptide-positive cells were found after nerve injury. Galanin- and galanin message-associated peptide-like immunoreactivity were found in 2% and 3%, respectively, of the Fluoro-Gold-labelled cell population normally and in 22% and 14%, respectively, after injury. No neuropeptide Y-positive cells were found in the Fluoro-Gold-labelled cell population normally, but after nerve injury, 96% of this population became neuropeptide Y-positive. Nitric oxide synthase-like immunoreactivity was found in 2% of the Fluoro-Gold-labelled cells normally and in 10% after injury. Two per cent of the Fluoro-Gold-labelled cells in the normal cases were stained by Griffonia (bandeiraea) simplicifolia lectin I -isolectin B4. After injury, however, no such double labelling was found. Thirty four per cent of the Fluoro-Gold-labelled cell population was carbonic anhydrase positive normally, and 42% after injury. Seventy-five per cent of the Fluoro-Gold labelled cells showed RT97 immunoreactivity normally and 12% after injury. Choleragenoid-like immunoreactivity was found in 99% of the Fluoro-Gold-labelled dorsal root ganglion cells normally and 81% after injury. Immunohistochemical visualisation of choleragenoid transganglionically transported from the injured sciatic nerve combined with neuropeptide Y immunocytochemistry showed that primary afferent fibres and terminals in the nucleus gracilis contain neuropeptide Y following peripheral nerve transection. Taken together, the results indicate that peripherally axotomised nucleus gracilis-projecting neurones undergo marked alterations in their cytochemical characteristics, which may be significant for the structural and functional plasticity of this system after injury. PMID- 7498391 TI - Zinc-containing telencephalic connections to the rat striatum: a combined Fluoro Gold tracing and histochemical study. AB - The organization of telencephalic zinc-containing neurons projecting to the rat striatum was investigated by combining intrastriatal injections of the retrograde fluorescent tracer Fluoro-Gold with histochemistry revealing zinc-containing neurons and terminals. Throughout the ipsilateral and contralateral neocortex, corticostriatal zinc-containing neurons with striatal projections were located predominantly at the border between deep layer V and superficial layer VI. Additional, but fewer zinc-containing neurons were located in layers II, III and deep layer VI of the ipsilateral neocortex. The main neocortical source of zinc containing afferents to the striatum were the frontal motor cortices. Smaller contingents of zinc-containing projections arose from the motor cortical forelimb and hindlimb areas and the parietal cortical areas. In the cingulate cortex, zinc containing neurons with striatal projections were found predominantly in the ipsilateral layers II and III, with only few neurons in the ipsilateral layer VI and in the contralateral layers II, III and VI. Subcortically, zinc-containing neurons belonging to the amygdalostriatal projection were found bilaterally in the basolateral and basomedial nuclei of the amygdala. Zinc has been found to modulate the response of many ligand- and voltage-gated ion channels, including both GABA receptors and NMDA-, AMPA- and kainate-type glutamate receptors. The present findings raise the possibility that zinc in the corticostriatal projections might play a role in the selective, possibly excitotoxic, cell death of GABAergic projections seen in Huntington's disease. PMID- 7498392 TI - Dye coupling between dorsal raphe neurones. AB - Neurones in the dorsal raphe nucleus (DRN) were impaled and filled with biocytin in coronal slices of midbrain taken from young adult rats. The electrophysiological properties and gross morphology of the cells were similar to those reported previously for serotonergic neurones in the DRN. Of 27 cases in which filled neurones were recovered in histological material, almost half (48%) showed labelling of two or three cells, although only one cell had been recorded from. Coupled cells were identified as close or distantly coupled, depending on the distance from the soma of the presumed impaled cell (23.5 +/- 15 microns, n = 7 and 150 +/- 26.5 microns, n = 10 respectively). Whereas close-coupled cells may have been artefactually "coupled" by the penetrating electrode, coupling between distant cells is most likely to be a result of transfer of biocytin through gap junctions. Camera lucida reconstructions of pairs of labelled cells revealed extensive overlap of dendritic fields and numerous crossings between dendrites. When examined at high magnification under a light microscope, many of the crossing dendrites were found to travel in different focal planes. Nevertheless, for each pair of cells, at least one point of close apposition was observed between dendrites or between the axon and a dendrite of the presumed impaled and coupled cell. The incidence of dye coupling between neurones in the DRN may reflect a relatively high level of electronic coupling between the neurones. This form of coupling may be important in determining the synchronous nature of firing of neurones in the DRN. PMID- 7498394 TI - Fractal correlation in human H-reflex. AB - The amplitude of the H-reflex has been known to have considerable variability even if the intensity of the stimulation is held constant. However, previous studies largely ignored the time-dependent profile of this variability. Recent mathematical analyses have shown that some seemingly irregular biological signals have fractal properties. A fractal time series is characterized by the property of self-similarity (self-affinity), and has long-range time correlation. The aim of this study was to investigate the question of whether the fluctuation of H reflex was fractal with strong time-correlation. Soleus H-reflexes were evoked in five healthy subjects at two levels of stimulation intensity [1.2 MT (motor threshold) and 0.9 MT] every 1 s and 1050 successive amplitudes of H-wave and M wave were recorded twice. The sequences of the H-wave and the M-wave amplitudes were analyzed by "coarse graining spectral analysis" to calculate the percentage of random fractal components in the sequences (%Fractal) and the spectral exponent beta. The %Fractal values of both sequences were above 90% [H-wave: 93.3 +/- 2.3% (1.2 MT), 91.6 +/- 3.1% (0.9 MT); M-wave: 94.3 +/- 3.3%; mean +/- SD]. Nonflat power spectra of the fractal components were observed for the H-wave sequences regardless of the stimulation intensity [beta = 0.75 +/- 0.26 (1.2 MT), 0.80 +/- 0.39 (0.9 MT)], indicating that the sequences were strongly time correlated. On the other hand, the M-wave sequences had a flatter spectrum (beta = 0.26 +/- 0.14) which was close to uncorrelated white noise. These results indicated that: (1) the fractal correlation found in the H-wave sequences was caused neither by the conduction through nerve fibers nor by the transmission at the neuromuscular junction, because the M-wave sequence had a significantly weaker time correlation, and (2) antidromic impulses in a motor nerve induced by the stimulation made a minor contribution to the generation of fractal correlation in the H-wave sequences, because it was preserved when the stimulation intensity was below MT. It was suggested that the fractal correlation in human H-reflex was generated at the synaptic connections to alpha-motoneurons in the spinal cord. PMID- 7498395 TI - Early and late stretch responses of human foot muscles induced by perturbation of stance. AB - In eight subjects standing on a movable platform, surface EMG activity was recorded from the foot muscles extensor digitorum brevis (EDB) and flexor digitorum brevis (FDB) and from the leg muscles soleus (Sol) and tibialis anterior (TA) during perturbations of upright stance. Perturbations inducing foot dorsiflexion (upward tilt and backward translation) evoked a short-latency response (SLR) and a medium-latency response (MLR) to stretch in the physiological extensors FDB and Sol, and a long-latency response (LLR) in the physiological flexors EDB and TA. Perturbations inducing plantar-flexion (downward tilt and forward translation) evoked the MLR in EDB and TA, and the LLR in FDB and Sol. The latency of the FDB and Sol SLR was compared to that of the H and T reflexes evoked in the same muscles by electrical or mechanical stimulation, respectively. In both muscles, the T reflex and the SLR followed the H reflex at delays accounted for by the different stimulation mode, indicating that the SLR induced in both muscles by upward tilt and backward translation was a true autogenetic stretch reflex from spindle primaries. The time interval between the onset of SLR and of MLR was significantly greater for the FDB than the Sol muscle, suggesting that MLR is a spinal reflex travelling through slower peripheral afferent pathways than SLR. From these latency differences and from the distance between the muscles, we calculated in four subjects the conduction velocity of the afferent fibres presumably responsible for the MLR in FDB. This was about 29 m/s. LLRs were evoked in TA and EDB during upward tilt and backward translation, and in Sol and FDB during downward tilt, but not forward translation. LLRs did not adhere to a proximal-to-distal pattern, since these could appear earlier in the foot than in the leg muscles. All responses were modulated by perturbation type (tilt vs translation) and body posture (normal stance vs forward leaning). Both the large amplitude of the foot muscle responses and their temporal pattern indicate that the muscles acting on the toes play a major role in stabilising posture. Their action increases in amplitude and extends in time the foot-ground reaction force, thereby improving the efficiency of the superimposed action of the leg muscle responses. PMID- 7498393 TI - In the hippocampus in vivo, nitric oxide does not appear to function as an endogenous antiepileptic agent. AB - Using a reverberatory epilepiform discharge of hippocampal-parahippocampal circuits termed "maximal dentate activation", this study investigated whether the local release of nitric oxide within these circuits functions as an antiepileptic agent. Two nitric oxide synthase inhibitors (L-nitro-arginine methyl ester and 7 nitro-indazole) and a guanylate cyclase inhibitor (methylene blue) were tested, and none had a significant effect on the time to onset or duration of maximal dentate activation. A membrane-permeable analogue of cyclic guanosine monophosphate (cGMP), 8-bromo-cGMP, caused an increase in the time to onset and a decrease in the duration of maximal dentate activation. The number of neurons expressing NADPH diaphorase activity (a marker for nitric oxide synthase) was also examined after repeated elicitation of maximal dentate activation. After 18 seizures there was a significant, but transient, decrease in the number of hilar/subgranular neurons that were NADPH diaphorase-positive. The decrease was only seen at 1 h after the last seizure. There was no induction of NADPH diaphorase activity. These results are not consistent with the hypothesis that, in hippocampal-parahippocampal circuits in vivo, nitric oxide is released in response to neuronal activity and then acts to terminate the neuronal activity. PMID- 7498397 TI - Surface electrodes are not appropriate to record selective myoelectric activity of splenius capitis muscle in humans. AB - Splenius capitis (SPL) electromyograms were recorded using conventional surface and intramuscular wire electrodes simultaneously during various head-neck movements and isometric tasks to test the selectivity of surface electrodes for SPL myoelectric signals. The insertion of bipolar wire electrodes was aided by a computerized tomographical study of each subject's neck. Surface electrodes were placed over the superficial SPL area. Head motion was recorded with an electromechanical device. The selective SPL wire recordings confirmed that SPL has two main functions: ipsilateral rotation and extension. It also plays a subordinate role in ipsilateral tilting of the head. Intramuscular and surface recording results were contradictory mainly for flexion and contralateral rotation. These discrepancies appeared to be due to 'cross-talk' from adjacent muscles, particularly from the sternocleidomastoid muscle. We conclude the validity of electrode recordings is questionable for SPL and most dorsal neck muscles, especially during isometric tests. PMID- 7498396 TI - Gating of sensation and evoked potentials following foot stimulation during human gait. AB - To investigate how gait influences the perceived intensity of cutaneous input from the skin of the foot, the tibial or sural nerves were stimulated at the ankle during walking or running on a treadmill. As compared to standing, the detection threshold for these stimuli was raised by more than 30% during the locomotion tasks. During walking, there was a phase-dependent modulation in perceived intensity of suprathreshold stimuli (1.5, 2, or 2.5 x PT). Stimuli given just prior to footfall were perceived as significantly above average (Wilcoxon signed-rank test). In contrast there was a significant phasic decrease in sensitivity for shocks delivered immediately after ipsi- and contralateral footfall. The amplitude of somatosensory evoked potentials (P50-N80 complex), simultaneously evoked from pulse trains to the sural nerve and recorded at scalp level, was, on average, 62% of the level during standing. During gait, the amplitude of this complex was significantly smaller just after footfall than the amplitude during late swing (MANOVA). It is suggested that the reduced sensation and the decreased evoked potentials after touchdown may be due to occlusion or masking by concomitant afferent input from the feet. On the other hand, the phasic increase in sensitivity at the end of swing is thought to result from a centrally generated facilitation of sensory transmission of signals in anticipation of foot-placing. PMID- 7498398 TI - Relative phase destabilization during interlimb coordination: the disruptive role of kinesthetic afferences induced by passive movement. AB - The disruption of three patterns of two-limb coordination, involving cyclical flexion-extension movements performed in the same or in different directions, was investigated through application of passive movement to a third limb by the experimenter. The three patterns referred to the homologous, homolateral, and heterolateral (diagonal) limb combinations which were performed in the sagittal plane. The passive movement involved a spatiotemporal trajectory that differed from the movements controlled actively. Even though subjects were instructed to completely ignore the passive limb movement, the findings of experiment 1 demonstrated a moderate to severe destabilization of the two-limb patterns, as revealed by analyses of power spectra, relative phase, cycle duration, and amplitude. This disruption was more pronounced in the homolateral and heterolateral than in the homologous effector combinations, suggesting stronger coupling between homologous than nonhomologous limb pairs. Moreover, passive mobilization affected antiphase (nonisodirectional) movements more than inphase (isodirectional) movements, pointing to the differential stability of these patterns. Experiment 2 focused on homolateral coordination and demonstrated that withdrawal of visual information did not alter the effects induced by passive movement. It was therefore hypothesized that the generation of extra kinesthetic afferences through passive limb motion was primarily responsible for the detriment in interlimb coordination, possibly conflicting with the sensory information accompanying active movement production. In addition, it was demonstrated that the active limbs were more affected by their homologous passive counterpart than by their nonhomologous counterpart, favoring the notion of "specific" interference. The findings are discussed in view of the potential role of kinesthetic afferences in human interlimb coordination, more specifically the preservance of relative phasing through a kinesthetic feedback loop. PMID- 7498400 TI - The dynamics of isometric bimanual coordination. AB - Eight right-handed subjects performed rhythmic isometric applications of torque in the directions of pronation and supination of the forearm, in single limb and bimanual conditions. Bimanual 'movements' were executed in either in-phase (homologous muscles simultaneously active) or anti-phase (non-homologous muscles active simultaneously) modes of coordination, in self-paced and frequency-scaled conditions. In the inphase (frequency-scaled) condition, subjects were required to synchronise (applications of torque) with each beat of a metronome, either in the direction of pronation or supination. In the anti-phase (frequency-scaled) condition, subjects were required to synchronise (applications of torque) with each beat of the metronome, either to the left or to the right. Departures from the anti-phase mode of coordination were observed as pacing frequency was increased. However, these departures were of short duration and the anti-phase mode was always re-established. These findings are in marked contrast to those obtained when there is free motion of the limbs. There also existed systematic differences between the stability of the pronation and supination phases of torque application. These differences were, in turn, modified through coincidence with the pacing signal. These results are discussed with reference to the constraints imposed upon the coordination dynamics by the intrinsic properties of the neuromuscular-skeletal system. PMID- 7498401 TI - Neurodegeneration in the central nervous system of apoE-deficient mice. AB - Apolipoprotein E (apoE) is involved in the development and regeneration of the central nervous system (CNS). ApoE may also be necessary to maintain the integrity of the synapto-dendritic complexity. We analyzed the synaptic alterations in the CNS of apoE-deficient (knockout) mice during the aging process. In apoE-deficient homozygous mice, there was an age-dependent 15 to 40% loss of synaptophysin-immunoreactive nerve terminals and microtubule-associated protein 2-immunoreactive dendrites in the neocortex and hippocampus, when compared to controls. Dendritic alterations were observed as early as 4 months of age. Ultrastructural analysis revealed extensive dendritic vacuolization and disruption of the endomembrane system and cytoskeleton in apoE-deficient homozygous mice. Further immunocytochemical studies of the neuronal cytoskeleton showed that in apoE-deficient mice there was a decrease in the immunoreactivity of alpha and beta tubulin (but not kinesin) in the cell bodies and processes. These results support the contention that apoE might play an important role in maintaining the stability of the synapto-dendritic apparatus and that altered or deficient functioning of this molecule could underlie the synaptic and cytoskeletal alterations in Alzheimer's disease. PMID- 7498399 TI - The stability of precision grip forces during cyclic arm movements with a hand held load. AB - In this paper we examine the coordination of grip force and load during brisk cyclic arm movements with a hand-held object under a range of conditions. We show that, regardless of the surface texture of the object or movement frequency, grip force is modulated in parallel with load. Thus, the tight coupling between grip force and load observed in short-duration tasks such as lifting or point-to-point movements is also seen in longer-duration cyclic movements. Moreover, the gain of the relation between grip force and load remains essentially constant over time. Across conditions, we find a dissociation between the gain of the relation between grip force and load and the grip force offset. With a more slippery surface texture both the gain and offset increase, whereas increases in frequency lead to an increase in the offset but a decrease in gain. This suggests that these two parameters are under independent high-level control. We also observe that when subjects were instructed to maintain a high-baseline grip force during the movement, grip force was still modulated with load even though an increase in grip was not necessary to prevent slip. This suggests that there is an obligatory coupling between grip force and load. This coupling might be subserved by low level mechanisms not under high-level control. PMID- 7498402 TI - Effects of exogenous nerve growth factor upon sympathetic sprouting into the hippocampal formation. AB - Following septal denervation of the hippocampal formation, sympathetic fibers from a transplanted superior cervical ganglion will innervate hippocampal tissue in a topographically restricted manner. Previously, we have shown a strong correlation between the regions innervated by the sprouting sympathetic axons and the distribution of NGF immunoreactivity in the hippocampal formation, suggesting that a restricted availability of NGF trophic support may regulate the topography of innervation by the NGF-sensitive sympathetic fibers. It was possible, however, that other molecular cues were responsible for restricting neurite outgrowth to selected hippocampal regions. In the current investigation, this idea was explored by experimentally altering the distribution of hippocampal NGF using a continuous intraparenchymal infusion device. Our results indicate that some hippocampal regions, not innervated by sympathetic fibers in control animals, do become occupied when the necessary trophic factor is provided, suggesting that these regions already contained an appropriate substratum for sympathetic neurite outgrowth but lacked the necessary trophic stimulus. Other regions, however, did not become innervated even though infused NGF was verifiably present in them. Together, these findings propose that a spatial restriction of NGF may be a crucial molecular mechanism for controlling the distribution of sprouting NGF sensitive sympathetic fibers but that other endogenous signals may regulate NGF's ability to stimulate local terminal sprouting. The data are also consistent with the idea that a limited availability of endogenous NGF is regulating both the density of sympathetic innervation into the hippocampal formation and the extent of neuronal survival within the transplanted ganglion. PMID- 7498404 TI - The effect of postinjury kindled seizures on cognitive performance of traumatically brain-injured rats. AB - The purpose of this experiment was to examine the consequences of postinjury seizures on cognitive performance after experimental traumatic brain injury (TBI). Rats either were injured at a moderate (2.1 atm) level of central fluid percussion TBI (n = 16) or were surgically prepared but did not receive a fluid pulse (sham-injured control, n = 16). Beginning 24 h after TBI, injured animals were injected (ip) once daily (Days 1-24 postinjury) with either saline (n = 8) or 25 mg/kg pentylenetetrazol (PTZ) (n = 8). Sham-injured rats were injected with an equal volume of saline (n = 8) or PTZ (n = 8). In both injured and sham injured animals, daily injections of PTZ resulted in an increase in the severity of behavioral seizures over days. On Days 25-29 after injury or sham injury, all animals were tested in the Morris water maze (MWM). Analysis of maze performance indicated that in sham-injured animals PTZ-produced seizures had a detrimental effect on performance. In injured animals, however, PTZ-treated animals exhibited significantly faster acquisition and better terminal performance in the MWM than did untreated injured animals. These results show that posttraumatic kindled seizures do not exacerbate behavioral deficits after TBI and may, in fact, improve recovery following injury. The findings of this experiment are consistent with the hypothesis that post-TBI neuronal depression may contribute to behavioral morbidity following injury. PMID- 7498403 TI - Lateromedial gradient of the susceptibility of midbrain dopaminergic neurons to neonatal 6-hydroxydopamine toxicity. AB - The topography-dependent vulnerability of midbrain dopaminergic neurons to neonatal intracranial exposure to 6-hydroxydopamine (6-OHDA) was investigated at adult age by the quantitative analysis of cell counts of tyrosine hydroxylase immunopositive neurons. In all cases of intracisternal 6-OHDA treatment, A9 dopaminergic neurons in the substantia nigra (SN) were much more vulnerable to death than more medially located A10 dopaminergic neurons. Moreover, within each cell group, there were also lateromedial topographic gradients. In the A9 neuronal group, cells located in the pars lateralis of the SN and the lateral part of the pars compacta of the SN were more susceptible to 6-OHDA toxicity than those located more medially. In the A10 neuronal group, cells located in the medial part of the ventral tegmental area were more resistant to toxicity than those located more laterally, and dopaminergic cells in the midline midbrain areas (interfascicular nucleus and rostral linear nucleus of raphe) were completely spared from 6-OHDA toxicity. These findings revealed that 6-OHDA is not equally toxic to all midbrain dopaminergic neurons in neonates and that the lateromedial vulnerability pattern shows similarities to those reported in Parkinson's disease. PMID- 7498405 TI - A new surgical technique that allows proximodistal regeneration of 5-HT fibers after complete transection of the rat spinal cord. AB - Shortening of the spinal column has been regarded as one possible method to obtain cord-to-cord apposition after total transection of the spinal column. However, to further improve regenerative possibilities, the problems of inconstant bony fusion and cyst formations within the junctions must be resolved. Modifying the method of de Medinaceli on the rat thoracic spine, we attempted several fixation devices to achieve better interspinal fixation after spondylectomy and transection, including transpedicular miniscrews, wiring of the transverse processes, and wiring of the posterior spinal processes. A dynamic model, based on retracting and compressing the cut ends of the spinal cord by means of adjustable fixation devices to allow swelling and shrinking of the stumps was also attempted to better compensate pathophysiologic changes of the transected cord. The best regeneration, as indicated by regrowth of 5-HT fibers below the level of transection, was obtained following application of fibrin glue and compressive wiring of posterior spinal processes. In this group, the distance between proximal and distal GFAP-rich spinal cord tissue (gap consisting of GFAP poor components such as cysts, phagocytic cells, and scar tissue) of the two spinal cord stumps was also the shortest. With better approximation, the numbers of regenerated 5-HT fibers improved remarkably, suggesting that this descending fiber system is able to bridge the transection under these conditions. PMID- 7498406 TI - Allele epsilon 4 of apolipoprotein E shows a dose effect on age at onset of Pick disease. AB - Pick disease is a rare progressive dementing illness characterized by severe atrophy of the frontal and temporal lobes. Clinically, Pick disease may be difficult to distinguish from Alzheimer disease (AD). The fact that Pick disease is often familial, and the evidence suggesting that the epsilon 4 allele of apolipoprotein E (ApoE) is a risk factor for AD and possibly other dementias, prompted us to study ApoE isoforms in Pick disease. ApoE genotypes were evaluated in an autopsy series of 21 AD and 12 Pick cases and compared with published data for a large group of adults participating in the Framingham Study. The distributions of ApoE genotypes in the AD and Pick patients and the controls were significantly different from one another. The frequency of epsilon 4 was 50.0, 20.0, and 13.6% in these respective groups. Linear regression analysis showed that the number of epsilon 4 alleles was inversely related to age at onset of Pick disease (P < 0.03) and accounted for 40% of the variation in age at onset. These results suggest that epsilon 4 may be a susceptibility factor for dementia and not specifically for AD. Experiments using a monoclonal antibody against ApoE suggest that neurons and Pick bodies are immunoreactive with ApoE. The dose effect of the epsilon 4 allele on age at onset of dementias other than AD and the association of ApoE immunoreactivity with neurons and Pick bodies support a broader role for ApoE in the pathogenesis of neuronal degeneration through interactions with the neuronal cytoskeleton. PMID- 7498407 TI - Increased expression of cathepsins E and D in reactive microglial cells associated with spongiform degeneration in the brain stem of senescence accelerated mouse. AB - Senescence-accelerated mouse (SAM) P8 and P10 exhibit a spongy degeneration, especially in the brain stem, and a brain atrophy mainly in the frontal portion of the cerebral cortex, respectively, with advancing age. In an attempt to clarify the role of two distinct intracellular aspartic proteinases, cathepsins E (CE) and D (CD), in these age-related pathological changes, accumulation and localization of these enzymes were investigated in the brain stem and the cerebral cortex of SAMP8 and P10 and in the senescence-resistant control SAMR1 with four different age groups (1 week and 2, 6, and 12 months). In the brain stem of SAMP8, a marked spongy degeneration was observed at more than 2 months of age. The same degree of spongy degeneration was also observed in the brain stem of age-matched SAMP10 but not SAMR1. The nonlysosomal enzyme CE was barely detectable in the brain stem of all three strains at 1 week of age, but it was markedly accumulated in the brain stem of SAMP8 and P10 at 2 months of age. The lysosomal enzyme CD was found in relatively high concentration in the brain stem of all three strains at 1 week of age. At 2 months of age, CD contents were significantly increased in the brain stem of SAMP8 and P10 compared with those of age-matched SAMR1. At the light-microscopic level, increased immunoreactivities for CE in the brain stem of 2-month-old SAMP8 and P10 were found in reactive microglial cells clustered at the spongy areas but not in microglial cells with resting or ramified morphology and astrocytes. The increased immunoreactivity for CD was observed mainly in reactive astrocytes and partially in reactive microglial cells. Immunoblotting analyses revealed that CE in the brain stem of 2 month-old SAMP10 consisted of only the mature form of 42 kDa, whereas CD in this tissue is composed of mainly the mature form of 44 kDa and partially its degradation products. On the other hand, there was a marked brain atrophy mainly in the frontal portion of the cerebral cortex of 6-month-old SAMP10 but not in age-matched SAMP8 or SAMR1. Although CE was not detectable even in the atrophied cortical area of SAMP10, CD contents in the cerebral cortex slightly increased with senescence in all three strains.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 7498408 TI - Differential macrophage responses in the peripheral and central nervous system during wallerian degeneration of axons. AB - We characterized quantitatively the macrophage response following axonal injury in both the peripheral (PNS) and central nervous system (CNS) of adult mammals. A monoclonal antibody (ED-1) which stains monocytes, macrophages, and activated microglia was employed. In one model, Wallerian degeneration of the sciatic nerve was studied. An increase in the number of macrophages was seen as early as 1 day following nerve transection. Macrophage number increased synchronously along the length of degenerating nerve over a 21-day period. In a second model, transection of a spinal dorsal sensory root allowed us to compare and contrast the macrophage response along the PNS and CNS portions of a single axonal pathway. An increased number of macrophages restricted to the PNS portion of this pathway was seen by 3 days and continued to increase over a 14-day period. Myelin breakdown occurred in association with an increase in the number of macrophages by 3 days in the PNS but not the CNS portion of the degenerating dorsal root axon pathway. Low affinity nerve growth factor receptor immunohistochemical staining increased by Day 1 in the PNS but not the CNS portion of this pathway, occurring prior to the invasion of macrophages. In both models, the morphology of infiltrating macrophages changed over time from small slender ramified cells to large elongated multivacuolated cells. In conclusion, our results demonstrate that the macrophage response during Wallerian degeneration of axons in adult mammals is much more rapid and robust in the PNS, where axonal regeneration occurs, than in the CNS, where axonal regeneration is far more limited. PMID- 7498409 TI - Decrease of neurotrophin-3 mRNA in adult rat hippocampus after pilocarpine seizures. AB - Neurotrophins such as NT-3 are subject to complex regulation during epileptic seizures. Pilocarpine at a dose of 250 mg/kg induced either limbic seizures (LS) or limbic motor status epilepticus (LMSE) in adult rats. In situ hybridization signals for NT-3 mRNA declined moderately after LS and were nearly lost in LMSE at 3-4 h. Loss of NT-3 mRNA expression does not correlate with cell death and may reflect a functional down-regulation in certain hippocampal neurons. PMID- 7498410 TI - c-JUN, KROX-24, and c-FOS expression in hippocampal grafts placed in excitotoxic hippocampal lesions of the rat. AB - Hippocampal transplants were examined for the expression of three specific immediate early gene encoded proteins in order to establish if grafted immature tissue maintains the basic molecular program necessary for gene transcription after grafting to excitotoxic (ibotenic acid-induced) lesions in adult host rat brains. The transplants were derived from newborn donor rats and were analyzed immunocytochemically for the presence of c-JUN, KROX-24, and c-FOS transcription factors 5 months after grafting. The expression and distribution patterns of these genes in the host hippocampus were identical to those in hippocampal neurons of normal untreated animals. c-JUN-, KROX-24-, and c-FOS-labeled neurons were also present in the transplants, where KROX-24 and c-FOS exhibited a distribution similar to host hippocampus. In contrast, c-JUN was more extensively expressed in the transplants, suggesting a molecular response to the grafting conditions and the actual graft-host brain interactions. PMID- 7498412 TI - Phagocytic cells in the rat olfactory epithelium after bulbectomy. AB - The phagocytic cells in the rat olfactory epithelium were examined after unilateral bulbectomy. Immunohistochemistry using a monoclonal anti-macrophage antibody (OX42) and electron microscopy showed that macrophages were present in the olfactory epithelium of the operated side 1 day after bulbectomy. The average number was 5.9 OX42-positive cells/2 mm septal epithelium, compared with 2.9 in unoperated controls. At 3 days the number of macrophages/2 mm epithelium peaked at 16 and subsequently dropped to 2.1 at 7 days. The number of macrophages and the timing of their appearance in the olfactory epithelium during the 1st week after bulbectomy were related to the acute and massive death of olfactory cells. The number of macrophages returned to control levels at longer postoperative times (14, 30, 60, and 90 days). Most of the OX42-positive macrophages were the ED1-positive activated type. At 6 and 7 days after surgery large phagosomes containing cell debris were observed in the cytoplasm of supporting cells in the epithelium. The phagocytosis by supporting cells was more evident at 30, 60, and 90 days, in the chronic stages of cell death. The results indicate that macrophages are the major phagocyte in the olfactory epithelium during the 1st week after bulbectomy and that both supporting cells and macrophages participate in phagocytosis at longer survival times. PMID- 7498411 TI - Glutamate-mediated excitotoxic death of cultured striatal neurons is mediated by non-NMDA receptors. AB - Considerable interest has focused on the role of glutamate-mediated excitotoxicity in neurodegenerative disorders of the basal ganglia. The in vitro data on the receptor mechanisms involved in this process, however, have been inconclusive. Some studies have indicated that excitotoxins acting at NMDA receptors kill striatal neurons and others have indicated that NMDA receptor mediated excitotoxic death of striatal neurons is minimal in the absence of cortex. In the present study, we used a pharmacological approach to carefully reexamine this issue in 2-week-old cultures of striatal neurons dissociated from E17 rat embryos. The sensitivity of these neurons to glutamate agonists and antagonists was determined by monitoring cell loss in identified regions of the growth dishes. We found that glutamate killed striatal neurons with an EC50 of 100 microM. This loss was not mediated by NMDA receptors, since it was not reduced by the NMDA receptor antagonist APV (0.1-1.0 mM). Consistent with this result, up to 50 mM NMDA receptor-specific excitotoxin quinolinic acid (QA) did not affect neuronal survival. Depolarizing the QA-exposed neurons with 35 mM potassium chloride to enhance NMDA receptor activation by QA also did not produce neuron loss. The metabotropic glutamate receptor antagonist AP3 (500 microM) also had no effect on the striatal neuron loss produced by 100 microM glutamate. In contrast, the non-NMDA antagonist GYKI 52466 (100 microM) did block the excitotoxic effect of glutamate (100 microM). Specific AMPA and KA receptor agonists and the non-NMDA antagonist GYKI 52466 revealed that the non-NMDA receptor-mediated excitotoxic effect of glutamate was mediated by KA receptors. These results suggest that cultured striatal neurons are directly vulnerable to non-NMDA glutamate agonists, but not to NMDA and metabotropic glutamate agonists. Thus, non-NMDA receptors may play a greater role in the excitotoxic death of striatal neurons in disease and experimental animal models than previously realized. PMID- 7498413 TI - Effects of nerve growth factor treatment on rats with lesions of the nucleus basalis magnocellularis produced by ibotenic acid, quisqualic acid, and AMPA. AB - Rats with bilateral lesions of the nucleus basalis magnocellularis (NBM) produced by ibotenic acid (IBO), quisqualic acid (QUIS), and alpha-amino-3-hydroxy-5 methyl-4-isoxazole propionic acid (AMPA) received either human recombinant nerve growth factor (NGF) (5.0 micrograms/day) or cytochrome c (cc) (0.3 microgram/day) treatment. Spatial memory impairments in the Morris water maze differed for the three toxins in the following order: IBO-cc > AMPA-cc > QUIS-cc. Treatment with NGF resulted in behavioral improvement for only the IBO-lesioned rats. Body weight was not affected by the different lesions before treatment; however, administration of NGF resulted in a decreased rate of body weight gain independent of the excitotoxin used. Reduction in choline acetyltransferase (ChAT) activity differed in the neocortex for the three toxins, whereas the hippocampus was unaffected. The cortical ChAT depletion was greatest for AMPA, intermediate for QUIS, and least for IBO. Restoration of ChAT activity by NGF differed for the three toxins (QUIS-NGF > IBO-NGF > AMPA-NGF), whereas ChAT activity in the hippocampus was increased equally. The loss of low-affinity NGF receptor (p75NGFr)-immunoreactive neurons differed for the toxins (AMPA-cc > QUIS cc > IBO-cc), whereas treatment with NGF increased the size of the remaining neurons independently of the used toxin. NGF induced a p75NGFr-immunoreactive thickening around intracerebral arteries for all three toxins. Lesions of the NBM produced by IBO, QUIS, and AMPA resulted in a quantitatively different pattern of behavioral, biochemical, and histological deficits. The quantitatively different pattern of recovery after administration of NGF may be determined by the responsivity of the remaining neurons. Intracerebroventricular administration of NGF affected noncholinergic systems as well, resulting in decreased food intake and cerebrovascular hyperinnervation. PMID- 7498414 TI - Interaction of apolipoprotein E with laminin increases neuronal adhesion and alters neurite morphology. AB - The extracellular matrix protein laminin profoundly affects neuronal adhesion, spreading, differentiation, and growth by binding integrin-type cell surface receptors. Laminin binds other basement membrane components, including heparan sulfate proteoglycans. Apolipoprotein E (apoE) also binds basement membrane and heparan sulfate proteoglycans and colocalizes with s-laminin in the neuromuscular junction. We sought to determine whether apoE interacts with laminin and whether, as a result, apoE alters the regulation of neuronal adhesion and differentiation by laminin. We demonstrate high-avidity interaction between apoE and laminin in vitro. Compared to laminin alone, a laminin-apoE substrate produces increased numbers of live, attached hippocampal neurons in culture. In addition, neurons grown on laminin-apoE substrates have larger growth cones, increased neuritic branching, and flattened cell bodies compared to neurons grown on laminin alone. ApoE may be important in the development and maintenance of neurons in the central nervous system by regulating interactions between the neuron and the extracellular matrix. PMID- 7498415 TI - Specificity of androgenic regulation of input to sexually dimorphic motoneurons in male rat lumbar spinal cord. AB - Averaged antidromic field potentials were recorded near motoneuronal somata of the spinal nucleus of the bulbocavernosus (SNB) after stimulation of their axons, in intact, castrated, and testosterone-treated castrated male rats under urethane anesthesia. When the SNB motoneurons were antidromically activated with double pulses to the bulbocavernosus (BC) nerve, the first pulse inhibited the antidromic response caused by the second pulse at interstimulus intervals of 5-40 ms, implying the presence of recurrent or other inhibition activated by BC nerve stimulation. Castration and androgen treatment had no significant effect on the amplitude of the suppression. In this regard this pathway differs from other pathways that synapse onto SNB motoneurons, in which activity is highly sensitive to androgen, indicating that androgen exerts its effects only on specific neural circuits that influence SNB motoneurons. PMID- 7498416 TI - (-)-nicotine protects against systemic kainic acid-induced excitotoxic effects. AB - (-)-Nicotine was shown to produce in vivo protection against neurobehavioral effects caused by systemically administered kainic acid (KA), an excitotoxin that has been widely used to induce temporal lobe convulsions including "wet dog shakes" in experimental animals. Rats pretreated with (-)-nicotine (0.5 mg/kg sc) 15 min before receiving KA (12.0 mg/kg sc) exhibited a marked reduction (P < 0.5) in the number of wet dog shakes when compared to saline-pretreated rats. Similarly, little visible brain damage was found in the (-)-nicotine-pretreated rats, but a widespread reduction in acetylcholinesterase-positive neurons was noted in the hippocampal areas of the saline-pretreated animals. While the mechanism of neuroprotection of (-)-nicotine is still not known, these findings suggest that (-)-nicotine may act as a therapeutic agent for putative excitotoxin mediated disorders. PMID- 7498417 TI - Intraperikaryal neurofilamentous accumulations in a subset of retinal ganglion cells in aged mice that express a human neurofilament gene. AB - Neurofilamentous changes in select groups of neurons are associated with the degenerative changes of many human age-related neurodegenerative diseases. To examine the possible effects of aging on the neuronal cytoskeleton containing human proteins, the retinas of transgenic mice expressing the gene for the human middle-sized neurofilament triplet were investigated at 3 or 12 months of age. Transgenic mice developed tangle-like neurofilamentous accumulations in a subset of retinal ganglion cells at 12 months of age. These neurofilamentous accumulations, which also involved endogenous neurofilament proteins, were present in the perikarya and proximal processes of large ganglion cells and were predominantly located in peripheral retina. The presence of the human protein may thus confer vulnerability of the cytoskeleton to age-related alterations in this specific retinal cell type and may serve as a model for similar cellular changes associated with Alzheimer's disease and glaucoma. PMID- 7498418 TI - Neonatal isolation enhances hippocampal dentate response to tetanization in freely moving juvenile male rats. AB - The impact of early neonatal isolation on measures of hippocampal neuronal plasticity was examined in freely moving male rats at 30 days of age. Beginning on Postnatal (PN) Day 2, one-half of pups from each experimental litter were individually isolated from the nest, dam, and siblings for a period of 1 h per day over PN Days 2-9, while their siblings remained in the nest. In addition, randomly selected litters served as unhandled controls. On PN Day 26 all pups were weaned and chronically implanted for recording of evoked field potentials and induction of hippocampal longterm potentiation. At 30 days of age, pups from the three treatment groups (isolated, nonisolated siblings, and unhandled controls) were tested for their ability to establish and maintain long-term potentiation across the perforant path/hippocampal dentate granule cell synapse. Changes in population EPSP slope and population spike amplitude (PSA) recorded following tetanization were used to assess the effects of neonatal isolation of hippocampal response measures. No significant between-group differences were obtained for input/output response curves constructed prior to tetanization. All three groups showed immediate and significant enhancement of the PSA measure at 15 min posttetanization. The level of PSA enhancement obtained from previously isolated pups was significantly greater than that obtained from both the nonisolated sibling and unhandled control groups.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7498419 TI - Human ventromesencephalic grafts restore dopamine release and clearance in hemiparkinsonian rats. AB - We and others reported previously that transplantation of fetal ventromesencephalic homograft restores the apomorphine-induced rotational behavior and electrochemical indices of dopamine (DA) depletion in the 6 hydroxydopamine (6-OHDA)-lesioned rat. We found that regeneration of KCl-evoked DA release and tyrosine hydroxylase (TH) immunoreactivity was limited to the graft area even 4 months after transplantation. In the present experiments, we transplanted human fetal ventromesencephalic tissue to the 6-OHDA-lesioned rats. After transplantation, rats received chronic cyclosporin and vibramycin treatment. We found that human fetal grafts from the substantia nigra can restore the effects of DA depletion in 6-OHDA-lesioned rats; these fetal grafts were found to reduce the apomorphine-induced rotational behavior and restore K(+) evoked DA release as well as DA clearance in the striatum. The area with active DA release is far beyond the transplantation site, unlike that seen in the homografted rats. These electrochemical responses correspond to the extended outgrowth of TH-positive neuronal fibers distal to the graft area. Taken together, our data suggest that rats that received human mesencephalic graft had much greater DA innervation and more complete restoration of function than those that received homografts. PMID- 7498420 TI - The Apyrase gene of the vector mosquito, Aedes aegypti, is expressed specifically in the adult female salivary glands. AB - The yellow fever mosquito, Aedes aegypti, expresses a gene, Apyrase (Apy), that encodes an ATP-diphosphohydrolase. The product of this gene is a secreted enzyme that facilitates hematophagy by preventing platelet aggregation in the host. Apy gene expression is limited to the cells of the distal-lateral and medial lobes of the adult female salivary glands. Apyrase protein levels, detectable by antibodies, peak in the salivary glands about 4 days after adult emergence and remain high after a blood meal. Primary sequence analysis of a genomic clone encoding apyrase reveals a unique TAAATA sequence and seven introns, as well as other conserved features of eukaryotic genes. The temporal, sex- and tissue specific expression of the Apy gene is consistent with its role as encoding a platelet anti-aggregation factor that functions to facilitate hematophagy and decrease probing time. PMID- 7498421 TI - Simulium damnosum s.l.: identification of inducible serine proteases following an Onchocerca infection by differential display reverse transcription PCR. AB - Vector-derived proteases are thought to be key to the regulation of filarial infections in Simulium damnosum s.I. To identify proteases of S. damnosum s.I. induced by infection with Onchocerca ochengi, a PCR-based differential display technique was used. By combining this method with homology-based serine protease primers transcripts can be detected from S. damnosum s.I. RNA. PMID- 7498422 TI - Induction of Trypanosoma cruzi metacyclogenesis in the gut of the hematophagous insect vector, Rhodnius prolixus, by hemoglobin and peptides carrying alpha D globin sequences. AB - Trypanosoma cruzi, a protozoan responsible for the American trypanosomiasis (Chagas disease), multiplies and differentiates in the gut of triatomine insect vectors. The effects of hemoglobin and synthetic peptides carrying alpha D-globin fragments on both the growth and the transformation of T. cruzi epimastigotes (noninfective) into metacyclic trypmastigotes (infective forms) were studied. This differentiation in the insect's gut is expressed when hemoglobin and synthetic peptides corresponding to residues 30-49 and 35-73 of the alpha D globin were added to the plasma diet. However, synthetic peptide 41-73 does not induce differentiation of epimastigotes even in the presence of the two former synthetic peptides. Thus, these data delineate an unusual molecular mechanism which modulates the dynamics of transformation of epimastigotes into metacyclic trypomastigotes in the triatomine vector's gut. PMID- 7498423 TI - Dermacentor andersoni: salivary gland proteins suppressing T-lymphocyte responses to concanavalin A in vitro. AB - Salivary glands obtained from feeding adult female Dermacentor andersoni (Acari:Ixodidae) were fractionated using differential centrifugation, detergents, centrifugal concentrators incorporating filter membranes with various molecular weight cutoffs, and preparative SDS-PAGE. A lymphocyte proliferation assay was used to evaluate the effects of salivary gland fractions on ConA-induced blastogenesis of normal murine splenocytes. Lipid, soluble, and detergent-soluble fractions were found to significantly suppress ConA-induced proliferation of splenocytes. Fractions containing soluble proteins suppressed splenocyte proliferation by ca. 26%. Suppressant activity in these fractions was due to components with molecular weights greater than 30 kDa. This suppression of splenocyte proliferation occurred with as little as 0.25 microgram protein per well. Salivary gland preparative SDS-PAGE fractions containing one or more soluble polypeptides or proteins with molecular weights in the range 36 to 43 kDa significantly suppressed murine splenocyte responses to ConA in vitro. PMID- 7498424 TI - Two genes encoding midgut-specific maltase-like polypeptides from Anopheles gambiae. AB - Full-length cDNA clones of two genes have been isolated from the African malaria vector mosquito, Anopheles gambiae. These genes, designated Agm1 and Agm2, encode maltase-like polypeptides of 498 and 599 residues, respectively. Deduced amino acid sequences contain a putative signal peptide sequence and four potential glycosylation sites. Agm1 and Agm2 show highest similarities to the Mal1 gene from Aedes aegypti and three clustered maltase genes from Drosophila melanogaster. Both genes are located at position 46D, in the terminal division of the left arm of the third chromosome. Agm2 has very strict tissue and temporal specificity, being expressed exclusively in the adult midgut. The specificity of Agm1 is similar but appears slightly broader; transcripts of this gene are detected at a low level in the pupae, and occasionally in the adult carcass after removal of the midgut. PMID- 7498425 TI - Wolbachia pipientis: bacterial density and unidirectional cytoplasmic incompatibility between infected populations of Aedes albopictus. AB - Unidirectional cytoplasmic incompatibility is seen when certain Wolbachia infected insect populations are crossed. Two hypotheses might explain this phenomenon: superinfections with mutually incompatible strains of Wolbachia producing incompatibility when crossed to individuals infected with only a single bacterial strain or, alternatively, a bacterial dosage model, with differences in Wolbachia densities responsible for the incompatibility. A quantitative PCR assay was set up as a general method to compare Wolbachia densities between populations. Using this assay in unidirectionally incompatible stocks of the mosquito Aedes albopictus, we have determined that densities are significantly higher in Houston than in the Mauritius and Koh Samui stocks. This is consistent with a dosage model for the observed crossing patterns, but does not rule out the possibility that superinfection is the primary cause of the incompatibility. PMID- 7498426 TI - Schistosoma mansoni: excretory-secretory polypeptides exhibit selective binding to plasma components of the snail Biomphalaria glabrata. AB - Previous studies have shown that Schistosoma mansoni excretory-secretory polypeptides (ESP) inhibit various internal defense functions of hemocytes from Biomphalaria glabrata and that plasma also may exert a modulatory effect on hemocyte activity. To better understand how plasma may influence hemocyte schistosome interactions in inbred strains of snails, biotinylated ESP (b-ESP) were used as probes to identify ESP-reactive plasma components and partially characterize the nature of their binding interactions. In a plasma binding assay, b-ESP bound in a dose-dependent fashion to immobilized snail plasma, although no quantitative differences in ESP reactivity to plasma of schistosome-susceptible (M-line) and -resistant (13-16-R1) snail strains were detected. Moreover, co incubation of b-ESP with homologous plasma or pretreatment of plasma with nonbiotinylated ESP in the plate assay significantly reduced b-ESP binding to immobilized plasma, indicating a specific interaction between ESP and plasma. Pretreatment of b-ESP with mannose, porcine stomach mucin, fetuin, and asialofetuin resulted in a significant inhibition of b-ESP binding to plasma, whereas pretreatment of immobilized plasma with a cocktail of monosaccharides (including mannose), porcine stomach mucin, and fetuin had no inhibitory effect. These data suggest the presence of a carbohydrate-binding protein(s) in sporocyst ESP that is targeted to plasma glycoconjugates. Based on the carbohydrate content of the major inhibitory glycoproteins, galactosyl and n-acetyl-galactosaminyl sugars may represent putative determinants for the lectin-like ESP molecule(s). However, ESP binding to plasma from M-line and 13-16-R1 B. glabrata strains exhibited similar sugar and glycoprotein inhibition patterns. SDS-PAGE and electroblot analyses further demonstrated that ESP bound to a subset of separated plasma polypeptides, most prominently to a doublet of 52 and 54 kDa, and a complex of proteins with molecular masses greater than 150 kDa. Other polypeptides exhibiting weaker binding interactions included components of 34, 60, 64, 86, and 125 kDa. Although both M-line and 13-16-R1 snail strains contained a similar complement of ESP-binding plasma proteins, ESP binding to the 34- and 86-kDa proteins occurred at higher frequencies in susceptible M-line plasma. It is concluded that ESP, released during in vitro transformation of S. mansoni miracidia, contain carbohydrate-reactive proteins capable of selectively binding to components of snail plasma. Quantitative differences between snail strains in the occurrence of ESP-binding plasma molecules was documented. PMID- 7498427 TI - Trypanosoma brucei and Trypanosoma cruzi: life cycle-regulated protein tyrosine phosphatase activity. AB - Recent evidence that Trypanosoma brucei synthesizes stage-regulated phosphotyrosine containing proteins and protein kinases stimulated us to assay bloodstream and insect stages of Trypanosoma cruzi and both pleomorphic and monomorphic clones of T. brucei for tyrosine phosphatase activity. Bloodstream and procyclic insect stages of T. brucei contained a 55-kDa protein that cross reacted with monoclonal antibodies directed against the human placental tyrosine phosphatase PTP1B. Protein lysates of all life cycle stages of both trypanosomes dephosphorylated a nonspecific substrate, pNPP, and the specific substrate Tyr(P)Raytide. Dephosphorylation of Tyr(P)Raytide was effectively inhibited only by sodium vanadate, a specific phosphotyrosine phosphatase inhibitor, but pNPP activity was also inhibited by sodium fluoride (NaF) in lysates of T. brucei and by NaF and sodium tartrate in lysates of T. cruzi, suggesting that their respective lysates also contained serine/threonine and acid phosphatase activities. Fractionation studies revealed that most of this activity was in the cytosol. Stage regulation of tyrosine phosphatase activity in T. cruzi was strongly suggested by differences in the optimal pH for tyrosine phosphatase activity (7.0 for amastigotes and epimastigotes; 5.0 for trypomastigotes). We conclude that both species of trypanosomes synthesize tyrosine phosphatases and propose that identification and characterization of the enzymes responsible for this phosphatase activity could provide information about trypanosomal virulence or the regulation of trypanosomal growth and differentiation. PMID- 7498428 TI - Trypanosoma brucei: molecular cloning of homologues of small GTP-binding proteins involved in vesicle trafficking. AB - Members of the rab subfamily of GTPases have been implicated as important components in vesicle trafficking in the eukaryotes. Individual rab proteins have a remarkable degree of specific subcellular localization. As a first approach to the study of these proteins in the protozoan parasite Trypanosoma brucei we have undertaken a survey of the rab subfamily using a strategy based on degenerate polymerase chain reaction utilizing the miniexon and the highly conserved WDTAGQE box which comprises part of the GTP-binding site. A number of T. brucei partial cDNAs were obtained from procyclic stage cDNA, and seven of these clones (designated rtb 1, 3-7, 9) were clearly members of the rab family. Northern analysis of poly(A)-selected RNA indicates that rtb1, 3, 4, and 7 are constitutively expressed at low levels in both life-stages of T. brucei. By Southern analysis of trypanosome genomic DNA and specific PCR from an isolated genomic clone, we show that two of the genes, RTB1 and RTB7, are adjacent or at least closely linked in the T. brucei genome, while the other five are dispersed. These data provide important molecular reagents for dissecting the unusual secretory pathway in this organism. PMID- 7498430 TI - Plasmodium gallinaceum: sporozoite invasion of Aedes aegypti salivary glands is inhibited by anti-gland antibodies and by lectins. AB - There is evidence which suggests that malaria sporozoites recognize mosquito salivary glands by specific receptor-ligand interactions. We are interested in identifying the putative salivary gland receptor(s) for sporozoite invasion. We used an in vivo bioassay for sporozoite invasion of salivary glands. In this assay, purified sporozoites from mature oocytes of Plasmodium gallinaceum were injected into Aedes aegypti mosquitoes and salivary glands were dissected at different time points after injection. One half of the maximum invasion of salivary glands by sporozoites occurred by 6 hr, and salivary gland sporozoite load did not increase further after 24 hr postinjection. This assay was used to determine the effect of experimental treatments with antibodies and lectins at 24 hr postinjection. We raised a rabbit polyclonal antiserum against female Ae. aegypti salivary glands which recognized tissue-specific determinants in the basal lamina of salivary glands. Purified IgG antibody fraction of the immune serum blocked sporozoite invasion in vivo. We tested a panel of 19 lectins and found 7 which bound to salivary glands. Of these 7, succinylated wheat germ agglutinin and wheat germ agglutinin completely blocked sporozoite invasion; Pisum sativum agglutinin and soybean agglutinin partially blocked; and concanavalin A, Dolichos biflorus agglutinin, and Phaseolus vulgaris erythroagglutinin did not block. Our results suggest that sporozoites interact with glycosylated salivary gland surface molecules which serve as receptors for invasion, and which may be in the salivary gland basal lamina. Because the putative sporozoite receptors contain immunogenic determinants, it is feasible to identify them by an immunological strategy. PMID- 7498429 TI - A factor Xa-directed anticoagulant from the salivary glands of the yellow fever mosquito Aedes aegypti. AB - Salivary gland extracts from the yellow fever mosquito Aedes aegypti contain a factor Xa-specific anticoagulant detectable in vitro by clotting assays. The anticoagulant is found only in female salivary glands, which is consistent with its role in bloodfeeding. Extracts of adult female glands significantly delay both the prothrombin time and the activated partial thromboplastin time, but have no effect on the thrombin clotting time. The anticoagulant has been shown to result from the specific inhibition of coagulation factor Xa based on a FXa clotting assay and the inhibition of FXa-directed cleavage of the synthetic chromogenic substrate, chromozym X. The inhibition of FXa by female salivary gland extracts exhibits noncovalent, noncompetitive inhibition kinetics and is reversible. The anticoagulant is destroyed by boiling for 10 min or heating at 56 degrees C for 30 min, has a 4 mM calcium optimum with no magnesium requirement, and has a pH optimum of 8.0. The anticoagulant activity has an apparent molecular weight of 35.5 kDa, as determined by molecular sieve. We propose that the A. aegypti anticoagulant is a novel, proteinaceous serine protease inhibitor specific for FXa. PMID- 7498431 TI - Onchocerca spp: a "family" of secreted acidic proteins expressed by infective larvae in blackflies. AB - Biosynthetic labeling of developing larvae of Onchocerca in blackflies has been used to characterize a group of stage-specific, secreted proteins produced by vector-stage parasites. These are highly acidic molecules (pI 4.4-5.1) present in at least three members of the genus (O. volvulus, O. lienalis, O. ochengi) that exhibit minor heterogeneity among species in apparent molecular mass (between 18 and 23 kDa). In O. volvulus, there are two polypeptides that run as a doublet of 18 and 20 kDa. In O. lienalis and O. ochengi, single polypeptides of 23 and 20 kDa were detected. The processes of synthesis and secretion appear to be temperature-sensitive and dissociated events. Experiments with O. volvulus in Simulium damnosum sl revealed that synthesis is initiated in second stage larvae and increases in infective-stage parasites: Secretion occurs when larvae leave the vector and enter the phase of development associated with the vertebrate host. Third-stage larvae of O. lienalis were shown to continue to express and accumulate the 23-kDa protein with age. The primary organ of secretion, as indicated by dissection, was the glandular esophagus. These data point to an important biological role for this group of molecules and suggest that they may belong to a family of related products. Because they have the distinctive characteristics of being secreted larval acidic proteins, we propose the acronym SLAP pending further insights into their functional properties. PMID- 7498432 TI - Aedes aegypti: a quantitative trait locus (QTL) influencing filarial worm intensity is linked to QTL for susceptibility to other mosquito-borne pathogens. AB - Because intensity of infection was significantly increased in a substrain of Aedes aegypti selected for susceptibility to the filarial worm, Brugia malayi, experiments were designed to assess numbers of microfilariae (mf) ingested and midgut penetration by mf in this susceptible substrain as compared to a refractory substrain selected from the same parental stock. Refractory mosquitoes ingested significantly fewer mf than susceptible mosquitoes and significantly fewer numbers of mf penetrated through refractory midguts as compared to susceptible midguts. In 16.7% of the refractory midguts, no mf were able to penetrate the midgut and in three refractory mosquitoes over 250 mf were ingested, but no mf penetrated the midgut. These results indicate that permissiveness of the midgut for penetration by microfilariae can determine not only parasite intensity, but also prevalence of infection. The genetic basis for ingestion of mf and midgut penetration was assessed using restriction fragment length polymorphism markers and quantitative trait loci (QTL) mapping. This mapping identified a QTL on chromosome 2, idb[2,LF181] (idb, intensity determinant for Brugia), that seems to influence ingestion ability. This QTL is linked to a previously identified QTL for susceptibility to B. malayi, fsb[2,LF98], as well as to loci for susceptibility to the malaria parasite, Plasmodium gallinaceum, and yellow fever virus. These results suggest that this region of chromosome 2 contains one or more genes that influence susceptibility of A. aegypti to several mosquito-transmitted pathogens. PMID- 7498433 TI - The black fly Simulium vittatum trypsin gene: characterization of the 5'-upstream region and induction by the blood meal. AB - The events leading to blood digestion in hematophagous insects are not well understood at the molecular level. Here we report on the characterization of a trypsin gene from the black fly Simulium vittatum. Southern blot analysis indicated that the S. vittatum genome contains at least two trypsin genes. Primer extension experiments identified two closely spaced transcription initiation sites, 3 bp apart. A genomic clone containing the trypsin gene was restriction mapped and the nucleotide sequence of the 5'-upstream region was determined. Several sequence elements were identified, including a consensus TATA box 35 nucleotides upstream from the transcription initiation site, a consensus arthropod initiator site at position -7, and two conserved sequence elements, 5'GGATTAA (position -77) and 5'TGTTTCCT (position -148). The latter two elements are present at comparable distances from the transcription initiation sites in black fly and mosquito trypsin genes. Significant amounts of trypsin mRNA were detected in guts of sugar-fed flies. Upon blood feeding, trypsin mRNA levels gradually increased to reach twice the initial abundance at 8 hr after blood ingestion. In contrast to other hematophagous insects, the induced mRNA level persists for a prolonged period of time (at least 48 hr) after blood ingestion. The results of this study provide information that may be useful for the expression of transgenes in insects of medical importance. PMID- 7498434 TI - Constitutive and blood meal-induced trypsin genes in Anopheles gambiae. AB - Trypsin genes in Anopheles gambiae are arranged as a tightly clustered gene family consisting of seven related coding sequences, devoid of introns. The two blood meal-inducible members of this family, Antryp1 and Antryp2, were shown to play a crucial role in the breakdown of the blood meal constituents. The role of Antryp3,4,5,6, and Antryp7 in the process of blood meal digestion remains to be elucidated. We have examined the localization and the expression patterns of these trypsins as well as the functional interactions in blood meal digestion between trypsins and other gut-specific proteases. Northern blot and RT-PCR analysis indicated that the genes Antryp3,4,5,6, and Antryp7 are all constitutively expressed in unfed female mosquitoes. Soon after blood feeding the mRNA of these trypsin genes became undetectable and appeared again at the end of the gonotrophic cycle. The blood meal-inducible trypsin Antryp1 was also constitutively expressed at low level in the gut of adult female mosquitoes. This trypsin was the only member of this gene family to be expressed in the gut of male and female pupae. By using antisera that specifically recognized recombinant Antryp4 we were able to show that the corresponding protein in Anopheles is synthesized and stored in the gut epithelium of unfed females as zymogen. Secretion and activation of this trypsin was shown to occur in the midgut lumen immediately after fluid ingestion and independently of the protein content of the meal. Recombinant trypsins expressed in Escherichia coli, with the exception of Antryp5 and Antryp6, were able to activate in vitro recombinant A. gambiae chymotrypsinogen, thus suggesting that blood meal ingestion is able to trigger a cascade of events leading to the activation of several proteases. PMID- 7498435 TI - Plasmodium gallinaceum: mosquito peritrophic matrix and the parasite-vector compatibility. AB - Transmission of malaria parasites occurs by relatively few species of mosquitoes. One proposed mechanism of refractoriness is an inability of certain Plasmodium spp. to cross the peritrophic matrix (PM) in the midgut of an incompatible mosquito. We have tested this hypothesis by studying sporogonic development of Plasmodium gallinaceum in susceptible (Aedes aegypti and Anopheles gambiae G3) and refractory (Anopheles stephensi) mosquito species in the presence and absence of the PM. In the presence of the PM the number of oocytes that developed in A. gambiae G3 was about 20% of that in A. aegypti, whereas no oocysts developed in A. stephensi. To disrupt PM formation we added, to an infectious bloodmeal, either exogenous fungal chitinase or polyoxin D, the latter being a potent inhibitor of chitin synthase. The absence of the PM did not increase the susceptibility of A. aegypti and A. gambiae nor did it make A. stephensi susceptible to P. gallinaceum infection. The data indicate that the PM is not the primary determinant of P. gallinaceum compatibility in these mosquitoes and suggest that determinant(s) of refractoriness occurs after the parasite crosses the mosquito PM. PMID- 7498436 TI - The expression of chloramphenicol acetyltransferase in mosquitoes and mosquito cells using a packaged Sindbis replicon system. AB - Sindbis (SIN) replicon virus was used to express chloramphenicol acetyltransferase (CAT) in Aedes albopictus (C6/36) cells and Aedes triseriatus mosquitoes. RNA transcribed in vitro from a SIN replicon plasmid (pSINrep5/CAT) and from SIN virus helper constructs (pDH-EB or pDH(26S)5'SIN) was coelectroporated into BHK-21 cells to generate replicon viruses, designated rep5/CAT/EB and rep5/CAT/26S. C6/36 cells infected with rep5/CAT/EB or rep5/CAT/26S virus at a multiplicity of infection of 3, expressed 3.8 x 10(6) and 6.0 x 10(6) CAT trimers per cell, respectively, at 2 days postinfection (pi). Both viruses attained peak titers by Day 2 pi. Adult female A. triseriatus mosquitoes were intrathoracically inoculated with 7 x 10(4) IFU rep5/CAT/EB or 1 x 10(5) IFU rep5/CAT/26S virus. Virus titers remained at approximately 10(5) IFU/ml through Day 2 pi and decreased roughly 1 log by Day 10 pi. CAT enzyme activity was detected 2 days pi (rep5/CAT/EB, 1.49 x 10(-4) units CAT/10 micrograms protein; rep5/CAT/26S, 2.03 x 10(-5) units CAT/10 micrograms protein) and remained near these levels through Day 10 pi. CAT was detected in the head, salivary glands, midgut, and ovaries of inoculated mosquitoes by indirect immunofluorescence or CAT activity assays. These results suggest that packaged replicon viruses can be useful for expression of heterologous genes in mosquito cells and whole mosquitoes. PMID- 7498437 TI - Trypanozoon: infectivity to humans is linked to reduced transmissibility in tsetse. I. Comparison of human serum-resistant and human serum-sensitive field isolates. AB - The transmissibility of recent isolates of human serum-sensitive (HSS) and human serum-resistant (HSR) Trypanozoon was compared by transmission of 37 stocks through an inbred line of Glossina m. morsitans. As in previous studies maturation was found to be dependent on fly sex with males producing significantly greater proportions of salivary gland infections than females. HSS stocks were, however, 1.8 times more likely to mature to mammalian infective form than HSR stocks in male tsetse and 2.7 times more likely to mature than HSR stocks in female tsetse. Infectivity to man has apparently evolved at the expense of transmissibility in tsetse. The likelihood of sexual processes occurring in Trypanosoma b. rhodesiense in wild flies is discussed. PMID- 7498438 TI - Trypanozoon: infectivity to humans is linked to reduced transmissibility in tsetse. II. Genetic mechanisms. AB - Trypanozoon infections are less likely to mature in female tsetse than in males. Analysis of maturation data from 37 Trypanozoon isolates in Glossina m. morsitans showed that while the proportion of mature infections (salivary gland infections as a proportion of established midgut infections) varied from isolate to isolate, the proportion of mature infections in female flies was consistently smaller than the proportion in male flies. The log of the probability of maturation in females is, on average, twice the log of the probability in males (estimate of the ratio of the logged proportions is 2.09, 95% confidence interval (CI) 1.8 to 2.5). Human serum-resistant isolates were less likely to mature than human serum sensitive isolates (ratio of logged proportions maturing was 1.5, 95% CI 1.3 to 1.8, in both male and female tsetse). Data for four other trypanosome stocks show that the probability of maturation decreases as the maturation time (the delay between the infected bloodmeal and maturation) increases. The decrease is approximately exponential with twice the half-life in male flies compared to that in female flies (estimate of the ratio of the exponential parameters is 1.97, 95% CI 0.7 to 3.3). A model is proposed to explain these observations which assumes that product(s) from an X-linked gene(s) kills or otherwise prevents migrating parasites from establishing a mature infection. Longer maturation times are associated with a heavy penalty in terms of transmissibility as measured by the vectorial capacity. PMID- 7498439 TI - Leishmania infantum: sequence homology within minicircle classes regardless of geographical distance. PMID- 7498441 TI - In vitro development of Xenopus skin glands producing 5-hydroxytryptamine and caerulein. AB - The granular glands of amphibian skin synthesize and store a large amount of bioactive amines and peptides which are structurally similar to mammalian brain gut peptides. To investigate the development of peptide- and amine-producing cells in the granular glands, pieces of dorsal skin taken at various stages from Xenopus laevis tadpoles were cultured, and the contents of caerulein and 5 hydroxytryptamine (5-HT) were measured. When pieces of skin from tadpoles at stages 57 to 60 (Nieuwkoop and Faber stages) were cultured in a medium containing 10% fetal calf serum (FCS medium) or one containing FCS treated with charcoal (chFCS medium), the caerulein and 5-HT levels were increased for the six days of the incubation period. The caerulein content was lower in the chFCS medium than in the FCS medium. Addition of thyroxine to the chFCS medium had no significant effect on the caerulein content. These results show that the caerulein- and 5-HT producing cells of the granular glands can develop in a culture system with FCS- or chFCS-containing media, and suggest that FCS contains substances which are absorbed by charcoal and stimulate development of the amine- and peptide producing cells of the glands. In a preliminary search for correlation between caerulein and 5-HT synthesis, addition of 5-hydroxytryptophan (5-HTP), a precursor to 5-HT, to the FCS medium increased 5-HT content and, conversely, caused significant decrease in caerulein content, suggesting that accumulation of caerulein in the granular glands is influenced by the amount of 5-HT synthesis. These studies indicate that this culture system is a useful model for investigating the development of peptide- and amine-producing cells. PMID- 7498440 TI - Genetic knockouts in mice: an update. AB - Gene disruption technology in mammals, by homologous recombination in embryonic stem cells, is a powerful method to manipulate the mouse germ line. In the past decade it has produced a wealth of knowledge concerning neuronal development, neurodegenerative disorders and the roles of oncogenes, Hox genes and growth factors during development. A surprising variety of genes, however, have given unexpected and disappointing results. A gene/function redundancy theory proposed by many investigators to explain the unexpected results has been supported in certain cases by the generation of double knockout mice. Modification of the basic technology now allows the investigators to carry out a variety of manipulations including conditional or tissue-specific knockouts. This may provide a better opportunity in the future for the gene therapy approach to correct the genetic disorder. PMID- 7498443 TI - High and low affinity transport of L-arginine in rat brain synaptosomes. AB - The uptake of L-arginine into purified rat brain synaptosomes was investigated with respect to time and various concentrations of L-[3H]arginine. Specific uptake was found to be linear with time for up to 5 min of incubation at 37 degrees C. Electrolytes, including sodium chloride, potassium chloride, magnesium chloride and calcium chloride, inhibited uptake of 3 microM L-arginine, and the inhibitory effect increased with increased electrolyte concentration under constant osmolarity. It was found that L-arginine was transported into synaptosomes by two uptake components--a high affinity component (3.5 microM) and a low affinity component (100 microM). These two components were similar to the Ly+ system because of their extreme sensitivity to inhibition by L-lysine and L ornithine but were distinguishable from each other by kinetic analysis of the uptake data and by their relative sensitivity to inhibition by several amino acids. PMID- 7498442 TI - Regulation of L-valine absorption by opioids interacting with mu-receptors in rabbit ileum. AB - In intact tissue, [D-Ala2,MePhe4,Gly-ol5]enkephalin (10(-5) M; mu-ligand), diminished short-circuit current (Isc) and increased water, Na+ and Cl- net fluxes in vitro under open circuit conditions; it also inhibited L-valine absorption and L-valine-dependent variations of short-circuit current (delta Isc,val). Naloxone (10(-6) M) antagonized these effects. In the absence of the muscularis and myenteric plexus this enkephalin or morphine (mu-ligand) reduced Isc and delta Isc,val. These enkephalin effects occurred at different times. Different concentrations of enkephalin were tested for their effects on delta Isc,val. [D-Ala2,D-Leu5]enkephalin (mainly a delta-ligand) significantly decreased Isc but not delta Isc,val. The reduction of L-valine absorption does not depend on the effects on basal ion transport. Interaction of opioids with mu receptors located in the submucosal plexus and/or in the epithelial cell accounts for this reduction. This enkephalin effect seems to be at least partially under the control of the myenteric plexus. PMID- 7498444 TI - Effects of polysorbates and Cremophor EL on vascular responses in rat aorta. AB - The effects of Tween 20, Tween 80, and Cremophor EL, surface active agents which are used for dispersion of water-insoluble substances, on vascular responsiveness were investigated using rat aortic rings. In high concentrations all these agents produced persistent contractions in aortic rings independent of the presence of endothelium. These contractions were not influenced by inhibitors of known endogenous contractile mediators. Incubation with these agents caused a concentration-dependent inhibition of the endothelium-dependent relaxant responses to acetylcholine in intact precontracted aortic rings. Endothelium independent relaxations produced by sodium nitroprusside were not inhibited, but rather potentiated in the presence of Tween 80 (10(-1) ml/l). On the other hand, Tween 80 inhibited the contractile effects of 5-hydroxytryptamine, phenylephrine, and bradykinin significantly. The data suggests that these substances affect both endothelial cells and vascular smooth muscle. PMID- 7498446 TI - Structure of rat tail tendon collagen examined by atomic force microscope. AB - The Atomic Force Microscope (AFM) was used to inspect collagen fibrils deposited on mica sheets at different fibrillogenesis times. Collagen was obtained from rat tail tendon fibers. Various fibril forms were observed, together with the characteristic periodic intra-fibril structure (D-bands). The fibril thickness, width, D-band periodicity and depth were measured and the statistical distribution of these parameters at 1, 2, 5, 10 and 15 days of in vitro fibril formation time was calculated. The fibrils showed an increasing size with time, but the band interval measure remained stable. The band depth, after an initial increase, exhibited a relative steadiness. The results indicate that AFM offers, at low resolution, images qualitatively similar to those obtained with electron microscopy, but with less manipulation of the sample. A quantitative evaluation of collagen structural features in the nanometer scale is made possible by AFM. PMID- 7498445 TI - Protective effect of lidocaine in the experimental foot-and-mouth disease pancreatitis. AB - Experimental infection of mice with foot-and-mouth disease virus (FMDV) induces a necrotizing pancreatitis of the exocrinar portion of the organ. The lesions are characterized by vascular congestion, edema and interstitial polymorphonuclear leukocyte (PMN) infiltrates. When infected mice were treated with different amounts of lidocaine (a local anesthetic, chemically defined as a tertiary amide compound), reduction in intensity of the pancreatic necrosis and in the number of PMN were observed. Even though lidocaine could interfere with FMDV post replicative cytolytic mechanisms, it appears that protection against pancreatic necrosis is by attenuation of PMN presentation in the infected tissue. PMID- 7498447 TI - A simple diagnostic method for the differentiation of Tamm-Horsfall glycoproteins from healthy probands and those from recurrent calcium oxalate renal stone formers. AB - Tamm-Horsfall glycoproteins (THPs)* from healthy probands, and those from a majority of recurrent calcium oxalate renal stone formers, reveal different properties when analyzed using isoelectric focusing. The pl-values of THPs from healthy probands are approximately 3.5 while THPs from recurrent renal stone formers have pl-values between 4.5 and 6. The two groups of THPs exhibit completely different protein patterns in IEF. This proves the structural difference of these THPs. The differences in IEF analysis allow the differentiation between THPs from healthy probands and those from recurrent calcium oxalate stone formers. These differences could possibly be used as a simple diagnostic method for the recognition of recurrent calcium oxalate renal stone formers. PMID- 7498448 TI - Origin and identification of bacteria which produce kairomones in the frass of Acrolepiopsis assectella (Lep., Hyponomeutoidea). AB - The volatiles used by the parasitoid Diadromus pulchellus to find its host, the leek moth, are produced by the bacteria developing in the frass of the host larvae. The origin and the nature of these bacteria were investigated. Samples were taken from healthy leeks and from infested leeks in the field, as well as from the frass of larvae reared in the laboratory either on the host plant or on an artificial diet. The various species of bacteria identified were cultured in the presence of precursors of leek sulphur volatiles and their volatile emissions were analysed. Klebsiella oxytoca and various Bacillus, common decomposers of plant matter, were the principal species producing active volatiles which were alkyl disulphides. PMID- 7498449 TI - Antimitotic and antiviral activities of Kelletinin A in HTLV-1 infected MT2 cells. AB - Kelletinin A [ribityl-pentakis (p-hydroxybenzoate)] (KA), a natural compound isolated from the marine gastropod Buccinulum corneum, showed antiviral activity on the human T-cell leukemia virus type-1 (HTLV-1) and antimitotic activity on HTLV-1-infected MT2 cells. KA inhibited cellular DNA and RNA synthesis, without influencing protein synthesis, and interfered with viral transcription by reducing the levels of high molecular weight transcripts. Finally, the compound inhibited HTLV-1 reverse transcriptase in vitro. PMID- 7498450 TI - Identification of a major cytokinin in coconut milk. AB - A major cytokinin found in coconut milk was isolated by using the tobacco callus growth-promoting assay as a guide during purification. The structure of the factor was determined to be 14-O-(3-O-[beta-D-galactopyranosyl-(1-->2)-alpha-D galactopyranosyl-(1-- >3)- alpha-L-arabinofuranosyl]-4-O-(alpha-L arabinofuranosyl)-beta-D- galactopyranosyl)-trans-zeatin riboside [G3A2-ZR] by various NMR techniques, including heteronuclear multiple bond connectivity by 2D multiple quantum NMR (HMBC), as well as mass spectroscopy and sugar analysis. The optimum concentration of G3A2-ZR for cytokinin activity in the tobacco callus assay was estimated to be 5 x 10(-6) M, so that G3A2-ZR is one order of magnitude more potent than 1,3-diphenylurea and one order less potent than zeatin riboside. At least 20% of the cytokinin activity of coconut milk could be attributed to G3A2-ZR. PMID- 7498452 TI - Report of the FAO expert technical meeting on the use of Hazard Analysis Critical Control Point (HACCP) principles in food control. PMID- 7498451 TI - Fibronectin peptide DRVPHSRNSIT and fibronectin receptor peptide DLYYLMDL arrest gastrulation of Rana pipiens. AB - Gastrulation is characterized by dramatic cell migration which is thought to require the interaction of cell adhesion molecules with extracellular molecules. We have tested two novel peptides, a fibronectin peptide and a fibronectin receptor peptide, for their effects on gastrulation of the leopard frog Rana pipiens. The fibronectin peptide DRVPHSRNSIT corresponds to residues 1373-1383 of the cell-binding domain of fibronectin; the receptor peptide DLYYLMDL corresponds to residues 124-131 of beta 1 subunit of a variety of integrins including alpha 5 beta 1. Either of these peptides significantly inhibited gastrulation after being microinjected into mid-blastulae. These results indicate that these sequences may correspond to the ligand/receptor interaction sites of fibronectin and its receptor(s). PMID- 7498453 TI - On the role of extracellular loops of opioid receptors in conferring ligand selectivity. AB - Based on an analysis of results taken from site-directed mutagenesis studies performed on opioid receptors, a role for the extracellular loops in conferring opioid subtype selectivity is proposed. It is suggested that the extracellular loop regions (which represent the region of highest sequence variability among opioid subtypes) interact with opioid ligands in a primarily non-specific fashion. Although these interactions are non-specific, they appear to play a discriminatory role in ligand binding and, in certain cases, prevent particular ligands from binding among receptor subtypes. We propose that selectivity may be imparted through a mechanism of exclusion, rather than specific pharmacophore recognition within the extracellular loops and N-terminal domain. This hypothesis is supported by a careful analysis of the binding profiles of several selective and non-selective ligands to a variety of chimeric mutants. These results, when combined with results taken from single-point mutation experiments point to the existence of a high affinity binding pocket within the transmembrane region which may be common among the opioid subtypes. PMID- 7498454 TI - Crystal structures of factor Xa specific inhibitors in complex with trypsin: structural grounds for inhibition of factor Xa and selectivity against thrombin. AB - Crystal structures of DX9065a and a related bisamidino-aryl inhibitor specific for the blood-clotting factor Xa have been solved in complex with bovine beta trypsin to a resolution of 1.9 A. Each inhibitor exhibits an extended conformation along the active site, in contrast to the compact folded structures observed for thrombin specific inhibitors. Few direct contacts (predominantly in the S1 pocket) are made between trypsin and the inhibitors. Transfer of the inhibitors to the active site of factor Xa suggests a three-site interaction: salt bridge formation at the base of the primary specificity pocket, extensive hydrophobic surface burial and a weak electrostatic interaction between the distal basic component of the inhibitor and an electronegative cavity of factor Xa formed by three backbone carbonyl oxygens. Additivity of these three interactions is the basis for the observed strong inhibition of factor Xa and provides a framework for the design of novel factor Xa inhibitors. A propionic acid group of the inhibitor would clash with the thrombin specific '60-insertion loop', thus conferring selectivity against thrombin. PMID- 7498455 TI - NMR identification of calcineurin B residues affected by binding of a calcineurin A peptide. AB - Triple resonance 3D NMR methods have been used to study the interaction between calcineurin B and a peptide fragment of calcineurin A for which it has high affinity (KD approximately 4 x 10(-7) M). Although calcineurin B aggregates at NMR concentrations of approximately 1 mM, in the presence of a target peptide fragment of calcineurin A it becomes monomeric and yields NMR spectra that are very similar to those reported previously for calcineurin B solubilized by the zwitterionic detergent CHAPS. Changes in chemical shifts between CHAPS- and peptide-solubilized calcineurin B are small which is indicative of no differences in secondary structure. Residues most affected by binding to target peptide are found primarily on the hydrophobic faces of the four helices, present in each of the two globular domains in calcineurin B, and in the loops connecting helices II and III, IV and V, and possibly in the C-terminal 12 residues, which also exhibit a change in mobility. PMID- 7498457 TI - Fast solute release from photosensitive liposomes: an alternative to 'caged' reagents for use in biological systems. AB - The kinetics of release of soluble marker trapped in liposomes of gel phase phospholipid containing a photoisomerisable phospholipid analogue have been investigated. Marker release is triggered by UV laser flash photolysis at 355 nm. A markedly temperature-dependent release rate is seen, and above 25 degrees C millisecond release kinetics can be achieved. These results suggest that such liposomes might find application as an alternative to conventional 'caged' reagents for photo-triggered reagent release in biological research. PMID- 7498456 TI - A photo-reactive derivative of ganglioside GM1 specifically cross-links VIP21 caveolin on the cell surface. AB - Previous studies have shown that sphingolipids may be enriched in caveolae, plasmalemmal invaginations implicated in endocytosis and signal transduction. We synthesised a radiolabeled derivative of ganglioside GM1 bearing a photo-reactive cross-linker at the end of its acyl chain. After insertion in the plasma membrane of cultured A431 or MDCK cells and photoactivation, the main protein cross-linked by the GM1 derivative was VIP21-caveolin, an essential structural component of caveolae. This result shows close proximity between GM1 molecules and VIP21 caveolin in the caveolar membrane and strongly implicates sphingolipid segregation in the biogenesis of caveolae. PMID- 7498458 TI - Occurrence and postmortem generation of anandamide and other long-chain N acylethanolamines in mammalian brain. AB - Long-chain N-acylethanolamines (NAEs) were assayed in pig, sheep and cow brain by gas chromatography/mass spectrometry of their tert.-butyldimethylsilyl derivatives in the presence of deuterium-labeled internal standards. Immediately after death, total NAEs ranged from about 2.7 micrograms/g wet weight (sheep, cow) to 6.5 micrograms/g wet weight (pig) and consisted almost exclusively (99%) of saturated and monounsaturated species. Anandamide (20:4n-6 NAE) comprised about 1% of total NAE in pig and cow brain, but was absent in freshly extracted sheep brain. When pig brain was analysed after 0.5, 1, 3, 4 and 23 h at ambient temperature, NAE levels were found to increase substantially over the entire time period with 20:4n-6 NAE formation exhibiting a time lag compared to that of saturated and monounsaturated NAEs. PMID- 7498459 TI - Cloning, sequencing and tissue distribution of two related G protein-coupled receptor candidates expressed prominently in human lung tissue. AB - A novel G protein-coupled receptor, named GPR12A, was cloned by a PCR strategy using degenerate primers designed from sequences conserved among receptors for inflammatory mediators. Screening of a human lung cDNA library with GPR12A as a probe also identified a closely-related cDNA (GPR6C.1) that has been previously reported as GPR4 [13]. GPR12A and GPR6C.1 are 46.1% identical in amino acid sequence, but are less than 33% identical to any other known receptors. Northern analysis revealed that they are expressed prominently in the lung. Although the ligands for GPR12A and GPR6C.1 are unknown, their similarity suggests that they are receptors for ligands of similar or identical chemical nature. PMID- 7498460 TI - A rapid and non-invasive selection of transgenic embryos before implantation using green fluorescent protein (GFP). AB - Non-invasive selection of transgenic mice was performed at the stage of preimplantation embryos. The morulae collected from wild female mated with hemizygous transgenic male expressing Aequorea victoria green fluorescent protein (GFP) under chicken beta-actin promoter could be classified as green or non-green under a fluorescent microscope. All the green embryos were shown to carry the transgene by PCR analysis. Taking advantage of the detection of GFP expression can be done non-invasively, the selected embryos were demonstrated to be able to developed to term with 100% of accuracy of the selection. PMID- 7498462 TI - Identification of the pore forming element of Semliki Forest virus spikes. AB - Pore formation at mildly acidic pH by SFV spike proteins was investigated using isolated and modified virions. Modification of the virions was performed by limited proteolysis in presence of octylglucoside and resulted in the formation of E1 particles and spikeless particles, respectively. Pore formation was detected by measuring the influx of propidium iodide into the viral particles. The results obtained clearly showed that the presence of E1 alone is sufficient to promote pore formation at mildly acidic pH. Thus E1 represents the pore forming element of the viral spike proteins. PMID- 7498461 TI - A P2X purinoceptor cDNA conferring a novel pharmacological profile. AB - We have cloned P2X4, a member of the P2-purinoceptor family, which has a new pharmacological profile. Rat P2X4 is distantly related to P2X1, P2X2 and P2X3 and is expressed in brain, spinal cord, lung, thymus, bladder, adrenal, testis and vas deferens. This ligand gated ion channel is activated by ATP and analogs with the potency order of ATP > ATP gamma S > 2-methylthio ATP > ADP approximately alpha beta-methylene ATP. However, none of the currently used P2X purinoceptor antagonists suramin, reactive blue 2 and PPADS blocked ATP evoked currents; in contrast their application resulted in potentiation of the agonist response. Due to lack of any known antagonist for P2X4 it is unlikely that native P2X4 has previously been recognized as a P2X purinoceptor. PMID- 7498463 TI - Self-association of CPV3, an avian thymic parvalbumin. AB - The avian parvalbumin called CPV3 readily forms disulfide-linked oligomers. Sedimentation data presented herein reveal that CPV3 also undergoes noncovalent self-association. Interestingly, the noncovalent interaction is promoted by either Ca2+ or Mg2+, whereas covalent complex formation displays an absolute requirement for the Ca(2+)-bound protein. Apo-CPV3 exhibits an apparent sedimentation coefficient of 2.08 S at 20 degrees C, in 0.15 M NaCl, 0.025 M HEPES-NaOH, pH 7.4. This value increases to 2.85 S or 3.16 S with addition of 1.0 mM Ca2+ or 5.0 mM Mg2+, respectively. Least-squares analysis of sedimentation equilibrium data suggests that 100 microM apo-CPV3 is primarily a mixture of monomeric and dimeric forms. With the addition of Ca2+, the equilibrium becomes exclusively monomer-trimer, with negligible amounts of dimer. A comparable distribution is observed in the presence of Mg2+. PMID- 7498464 TI - The peripheral cannabinoid receptor: adenylate cyclase inhibition and G protein coupling. AB - Two cannabinoid receptors, designated neuronal (or CB1) and peripheral (or CB2), have recently been cloned. Activation of CB1 receptors leads to inhibition of adenylate cyclase and N-type voltage-dependent Ca2+ channels. Here we show, using a CB2 transfected Chinese hamster ovary cell line, that this receptor binds a variety of tricyclic cannabinoid ligands as well as the endogenous ligand anandamide. Activation of the CB2 receptor by various tricyclic cannabinoids inhibits adenylate cyclase activity and this inhibition is pertussis toxin sensitive indicating that this receptor is coupled to the Gi/G(o) GTP-binding proteins. Interestingly, contrary to results with CB1, anandamide did not inhibit the CB2 coupled adenylate cyclase activity and delta 9-tetrahydrocannabinol had only marginal effects. These results characterize the CB2 receptor as a functional and distinctive member of the cannabinoid receptor family. PMID- 7498465 TI - Isolation and characterization of an arylalkylamine N-acetyltransferase from Drosophila melanogaster. AB - The enzyme arylalkylamine N-acetyltransferase (aaNAT) catalyzes the rate-limiting step in melatonin formation in the vertebrate pineal gland. Numerous attempts to purify this highly unstable enzyme from vertebrates have been unsuccessful. Here, we report the purification of an aaNAT enzyme from Drosophila melanogaster, using a radioenzymatic activity assay and column chromatography. The isolated 29.5-kDa protein acetylates tryptamine, dopamine and serotonin with affinities of 0.89 to 0.97 mM, respectively. This suggests that the identified aaNAT may be involved in melatonin synthesis and sclerotization as well as in neurotransmitter catabolism in insects. PMID- 7498466 TI - A novel approach to the design of potent bioactive peptides by incorporation of proline brackets: antiplatelet effects of Arg-Gly-Asp peptides. AB - Enhancing the potency of peptides is a critical and important step in the development of peptide drugs. We have proposed that proline residues flanking protein-protein interaction sites perform a structural role in enhancing their interaction [R.M. Kini and H.J. Evans, Biochem. Biophys. Res. Commun. 212 (1995) 1115-1124]. To test this theory, we incorporated proline residues on either or both sides of the interaction site of an antiplatelet peptide, IARGDMNA and determined the inhibitory potency of the peptides in whole blood aggregation. Inclusion of one proline residue, on either the amino or carboxy terminal side of the interaction site, enhances the antiplatelet activity to approximately the same extent (1.5- to 2.5-fold). Incorporation of proline residues on both sides enhances the activity by 7- to 13-fold. This enhancement of the biological activity of the peptide is probably due to a reduction in the number of possible conformations of the peptide, without introducing the rigidity that would accompany cyclization. Incorporation of proline brackets thus provides a novel approach to the design and development of more potent peptide drugs and ligands. PMID- 7498467 TI - The fastest actin-based motor protein from the green algae, Chara, and its distinct mode of interaction with actin. AB - The endoplasmic streaming in Characean cells is an actin-dependent movement. The motor protein responsible for the streaming was partially purified and characterized. It was soluble at low ionic strength, an ATPase of a molecular mass of 225 kDa and activated more than 100 times by muscle F-actin. Surprisingly, in an in vitro motility assay, the motor protein moved muscle F actin at 60 microns/s, which is similar to the velocity of streaming in a living cell and 10 times faster than muscle myosin. Proteolytic cleavage of actin impaired movement crucially on muscle myosin, but did not affect movement at all on the Chara motor protein, suggesting that the Chara motor protein would interact with actin via a set of sites different from those of muscle myosin. PMID- 7498468 TI - Sequence-specific DNA binding activity of rat cdc37-related gene product. AB - The rat cdc37-related gene product (RCdc37), which is possibly involved in the regulation of cell cycle progression, contains a putative basic/leucine zipper (bZIP) domain in its N-terminal portion. In this study, we have identified a rat genomic sequence which can interact with RCdc37 using an in vitro binding assay. The specificity of this interaction was confirmed by gel retardation experiments. These results raise a possibility that RCdc37 might play an important role in the control of cell cycle progression via a sequence-specific DNA binding mechanism. PMID- 7498469 TI - Hucolin, a new corticosteroid-binding protein from human plasma with structural similarities to ficolins, transforming growth factor-beta 1-binding proteins. AB - In order to study the cortisol-binding factors in blood, human plasma was applied to a 11 beta-hydroxy-3-oxo-4-androstene-17 beta-carboxyaminoethylamine-(HACA) 1,4 butanediol diglycidyl ether-Sepharose column. Elution of the column with cortisol buffer produced two protein peaks, the minor peak yielded a protein complex of molecular weight approximately 200 kDa, subsequently termed hucolin. SDS-PAGE analysis under reducing and non-reducing conditions revealed hucolin was a disulphide-linked complex of 35-kDa and 75-kDa subunits. Twenty-five amino acid residues of the N-terminus of the 35-kDa subunit were determined and homology searches revealed an 88% sequence identity with the N-terminal region of beta ficolin, a transforming growth factor-beta 1 (TGF-beta 1)-binding protein purified from porcine uterus. PMID- 7498470 TI - Amino acid residue: is it structural or functional? AB - A new approach is suggested for delineating the structural and functional amino acid residues in proteins with known three-dimensional structure, basing on the involvement of residues in intramolecular hydrophobic and hydrophilic interactions and additional information about the conservativity of the residues. The approach is applied to the families of homologous neurotoxins and cardiotoxins. The results obtained concerning the role of amino acid residues in both families of toxins accord well with the similarity of their fold, but different mechanisms of action. Current approach can be used for detailed characterization of protein spatial structures, as well as for rational protein engineering. PMID- 7498471 TI - Interaction of glyceraldehyde-3-phosphate dehydrogenase with SH-containing compounds: evidence for the binding of L-cysteine and for the dependence of the binding on the functional state of the enzyme. AB - Incorporation of L-[35S]cysteine into rabbit muscle glyceraldehyde-3-phosphate dehydrogenase was detected following incubation of the enzyme in a mixture containing glyceraldehyde-3-phosphate, NAD+ and the labeled cysteine. Insignificant binding occurred in the absence of either the substrate or NAD+, suggesting that formation of an acylated enzyme form was a prerequisite for the binding. Stoichiometry of the binding depended on the number of functioning active centers; up to 4 moles of L-[35S]cysteine bound per mole tetramer with fresh enzyme preparations. The L-[35S]cysteine incorporation depended on pH and was maximal when a group having pKa of 8.5 is protonated. To clarify the relevance of this finding to the effect of SH-containing compounds previously shown to decrease the rate of 3-phosphoglyceroyl-enzyme hydrolysis [Kuzminskaya et al., FEBS Lett. 336 (1993) 208-210], the pH-dependence of the effect of glutathione on the hydrolysis rate was determined and found to be close to the pH dependence of L-[35S]cysteine binding. PMID- 7498473 TI - Mechanism of Lac repressor switch-off: orientation of the Lac repressor DNA binding domain is reversed upon inducer binding. AB - Lac repressor's DNA-binding domains contain helix-turn-helix motif which, though similar to those of phage lambda Cro protein, are oriented differently with respect to DNA: in the specific complexes with Lac operator, N termini of the repressor's subunits are facing inwards. We demonstrate that, in the presence of an inducer, the repressor's N termini cross-link to the operator's outermost nucleotides. We suggest that the inducer fixes the repressor's DNA-binding domains in the Cro-type configuration and thus garbles its recognition surface. Since the Cro-type configuration is perfectly suitable for binding the DNA, this also explains how the switched-off repressor retains its non-specific DNA binding. PMID- 7498472 TI - Resistance of Ehrlich tumor cells to apoptosis can be due to accumulation of heat shock proteins. AB - Previously we have found that stationary Ehrlich ascites carcinoma (EAC) cells in vivo accumulated heat shock proteins (HSPs) and became resistant to necrotic death induced by prolonged energy deprivation of hyperthermia. Here we report that apoptotic death induced by nutrient starvation, transient ATP depletion, heat shock and a microtubule-disrupting drug, vinblastine, was also suppressed in stationary EAC cells comparing with exponential cells. When exponential (sensitive) cells were subjected to short-term heating with recovery to accumulate inducible form of HSP70, they also became resistant to all of the employed apoptosis-inducing exposures, and an inhibitor of cytosolic protein synthesis, cycloheximide, prevented acquisition of the resistance. It is suggested that in vivo accumulation of HSPs in stationary tumor cells can be endogenous protective device against apoptotic death induced by starvation or some anticancer treatments. PMID- 7498474 TI - Analysis of a novel defective HTLV-I provirus and detection of a new HTLV-I induced cellular transcript. AB - HTLV-I generally integrates at least one full-length copy in adult T-cell leukemia (ATL) cells. A group of patients without full-length provirus have a unique conserved truncation of the provirus which retains env-pX-3'LTR. Tumor cells of a patient from this group were genetically analyzed. Analysis of the 5' and 3' cellular flanking region adjacent to the provirus suggest that the defective provirus was integrated immediately downstream of a promoter of an unknown cellular gene. The activity of the promoter was weak but was responsive to Tax-like HTLV-I LTR. The provirus may have utilized it as a substitute for the 5'LTR and thus 3'LTR may have become an alternative promoter for the cellular gene, which may give similar viral-cellular interactions to that of general cases with full-length proviruses. Surprisingly, the 3' cellular flanking region which is thought to be controlled originally by the promoter is constitutively expressed specifically in an HTLV-I producing ATL cell line HUT1O2G, in which the corresponding region is not modified by provirus. The detection of this HTLV-I induced transcript provides a probe to find an HTLV-I inducible unknown cellular gene that may be related to the pathogenesis of ATL. PMID- 7498475 TI - rap1 p21 regulates the interaction of ras p21 with RGL, a new effector protein of ras p21. AB - We have recently found that ralGDS family members (RGL and ralGDS) are putative effector proteins of ras p21. rap1 p21 is a small GTP-binding protein which has the same amino acid sequence as the effector loop of ras p21. We examined the effect of rap1 p21 on the interaction of ras p21 with RGL. The GTP-bound form of rap1 p21 interacted with RGL as well as did ras p21. rap1 p21 inhibited the interaction of ras p21 with RGL. RGL was phosphorylated by cyclic AMP-dependent protein kinase (protein kinase A). Phosphorylation of RGL did not affect its binding to ras p21 and rap1 p21 under the conditions that phosphorylation of Raf 1 reduced its affinity for ras p21. These results demonstrate that rap1 p21 but not protein kinase A regulates the interaction of ras p21 with RGL and suggest that rap1 p21 and protein kinase A may cooperate to distinguish the signal or ras p21 to RGL from that to Raf-1. PMID- 7498476 TI - In vitro glycation of glomerular basement membrane alters its permeability: a possible mechanism in diabetic complications. AB - The permeability of glomerular basement membrane (GBM) was assessed in vitro by the filtration of solutions of proteins across films formed from isolated pig GBM. Incubation of the films with fructose or glucose increased their permeability to water and serum albumin. The effect of fructose was similar to that previously noted for films crosslinked with glutaraldehyde. The metal chelator DTPA abolished the effects of glycation; EDTA was partially effective in this respect. Transition metal catalysed formation of glycoxidation induced crosslinks may explain the increased permeability of glycated GBM. PMID- 7498477 TI - Inverse deuterium kinetic isotope effect for peroxidation in human low-density lipoprotein (LDL): a simple test for tocopherol-mediated peroxidation of LDL lipids. AB - alpha-Tocopherol (alpha-TOH) can act as a pro- or antioxidant for isolated ubiquinol-10-free human low density lipoprotein (LDL). We demonstrate that alpha TOH is a more potent pro-oxidant than other forms of vitamin E for LDL peroxidation initiated by mild fluxes of aqueous peroxyl radicals and low concentrations of Cu2+. A simple deuterium exchange test shows that alpha-TOH switches from pro- to anti-oxidant at Cu2+:LDL ratios > 2.5. The results suggest that this test may be useful to distinguish 'inhibited' peroxidation of emulsion lipids propagated via the lipid peroxyl radical from that mediated via the antioxidant radical. PMID- 7498478 TI - Predicted structure and possible ionmotive mechanism of the sodium-linked NADH ubiquinone oxidoreductase of Vibrio alginolyticus. AB - Two groups have now published sequences of the six genes contained in the operon coding for the sodium-linked NADH-ubiquinone oxidoreductase of Vibrio alginolyticus. Sequence analyses indicate that this enzyme is unrelated to other known respiratory NADH dehydrogenases. A search for cofactor motifs suggests that the enzyme contains only one FAD, a ferredoxin-type iron sulphur centre, and the NADH-binding site. These are all located on NqrF, a subunit that can be recognized as a new member of a large diverse family of NAD(P)H-oxidizing flavoenzymes. A possible model of ion-coupling is presented, based upon this new information. PMID- 7498479 TI - Genomic structure and alternative splicing of the murine bhk/ctk/ntk gene. AB - Recently, we and others have cloned cDNAs encoding a second member of the Csk family of inhibitory protein kinases, which we termed Bhk [M.A. Ershler et al. (1994) Dokl. Akad. Nauk. 339, 679-683]. In the present study, two new distinct types of bhk mRNA were found in addition to the third form described previously. Analysis of the bhk genomic structure established that three exons participate in the alternative splicing of bhk mRNA. PMID- 7498480 TI - Stimulation of synaptosomal free radical production by fatty acids: relation to esterification and to degree of unsaturation. AB - The ability of three fatty acids and their respective ethyl esters, to promote generation of reactive oxygen species (ROS), was compared in a preparation of rat brain synaptosomes. Arachidonic but not palmitic or linoleic acids promoted ROS generation. Ethyl esterification of each fatty acid significantly enhanced ROS production and also levels of lipid peroxidaton. Pro-oxidant activity was enhanced by fatty acids, proportionally to their degree of unsaturation. Since ethanol consumption is known to lead to esterification of membrane lipids, this transformation may in part account for the ROS-promoting potential of alcohol. PMID- 7498481 TI - Molecular cloning and characterization of a novel truncated from (ClC-2 beta) of ClC-2 alpha (ClC-2G) in rabbit heart. AB - Two cDNAs encoding rabbit heart Cl- channels (rabClC-2 beta and rabClC-2 alpha) were isolated by a PCR cloning strategy. RabClC-2 beta is a novel cDNA consisting of 2998 bp and encoding the 822-amino acid protein, while rabClC-2 alpha is identical to previously reported ClC-2G. RabClC-2 beta is 68 amino acids truncated from NH2-terminus of rabClC-2 alpha, but all 13 putative hydrophobic domains are conserved in rabClC-2 beta. Although rabClC-2 alpha was suggested to be activated by extracellular hypotonicity, expression of rabClC-2 beta in Xenopus oocytes induced large Cl- currents even in the absence of extracellular hypotonicity. Induction of external hypotonicity did not further increase the amplitude of membrane currents. On the other hand, as similar to rabClC-2 alpha, rabClC-2 beta current was augmented by PKA activation. Thus, different RNA processing of the same gene appears to provide two highly homologous PKA activated Cl- channels with or without responsiveness to cell swelling in rabbit heart. PMID- 7498482 TI - A novel chemoenzymatic glycosylation strategy: application to lysozyme modification. AB - Hen egg lysozyme has been non-specifically glycosylated using a novel two-step strategy. First, a number of sucrose molecules have been chemically bound to the protein surface lysines, then the glycosidic chains have been enzymically lengthened, using a glycosyltransferase. For this task, a fructosyltransferase and a levansucrase have been tested, the latter appearing as the most effective one. In all cases, reactions have been optimised and several degrees of modification have been obtained. Finally, the effects of the modifications on lysozyme hydrophobicity, hydrolytic activity, hydrolysis substrate affinity and thermostability have been assessed. PMID- 7498483 TI - Relationship between muscle myosin isoforms and contractile features in rabbit fast-twitch denervated muscle. AB - The effects of 8-day-old rabbit fast-twitch gastrocnemius denervation on the type of myosin isoforms and on contractile features (maximum velocity Vmax and contraction time (CT) of the muscle were followed between 15 and 60 days postnatal. The myosin isoforms and the Vmax and CT values of the denervated gastrocnemius displayed large changes during this period. These changes, which led at 2 months postnatal to a muscle displaying the properties of a slow-twitch muscle did not occur in synchrony: complete conversion to slow-type myosin isoforms occurred only at 60 days postnatal, whereas complete conversion to slow twitch Vmax and CT values occurred as soon as 35 days postnatal. The results address a new question concerning the relationship between muscle myosin and contractile features. PMID- 7498484 TI - Cellular distribution of protein kinase C isozymes in CD3-mediated stimulation of human T lymphocytes with aging. AB - Protein kinase C (PKC) is involved in a variety of cellular responses, such as the expression and secretion of IL-2, the regulation of cytotoxic killing and cell proliferation. It is known that these immune functions are altered with aging. Here, we show that anti-CD3-triggered T cell proliferation is significantly decreased with aging and that H7, an inhibitor of PKC, impairs the anti-CD3-induced T cell proliferation in a differential manner, lymphocytes of healthy young subjects being more sensitive to the PKC inhibitor than those of elderly subjects. We examined (Western blot) the presence and the cellular distribution of PKC isozymes in T lymphocytes of healthy young and elderly subjects in the resting state and after anti-CD3 mAb stimulation using antibodies directed against PKC alpha, beta, delta, epsilon and zeta isoforms in the cytosol and the plasma membrane fractions. These five PKC isotypes were present in human T cells of young and elderly subjects. However, their distribution between the cytosolic and membrane fractions varied according to the isozymes and the age of the subjects. In resting lymphocytes of young subjects, all the PKC isozymes were found in the cytosolic fraction, except PKC-zeta. In resting lymphocytes of elderly subjects PKC-zeta and -epsilon were almost equally distributed between the cytosolic and the membrane fractions, whereas PKC-alpha and -zeta were mainly found in the membrane fraction and PKC-beta was almost exclusively located in the cytosolic fraction. The translocation of PKC-alpha, -beta, -delta and -epsilon could be observed under anti-CD3 mAb stimulation in lymphocytes of young subjects, while in the case of elderly subjects only the PKC beta isoform was translocated. Our results suggest tha the decreased availability of cytosolic PKC may contribute to the diminished PKC-dependent responses to CD3-triggered stimulation of human T lymphocytes with aging. PMID- 7498487 TI - Temperature-induced rapid increase in cytoplasmic free Ca2+ in pathogenic Leishmania donovani promastigotes. AB - We demonstrate that the cytoplasmic free Ca2+ concentration in three virulent Leishmania donovani promastigote strains is maintained at 20-30 nM level whereas the avirulent promastigotes maintain [Ca2+]i at 80-100 nM. Rapid exposure of virulent promastigotes to higher physiological temperature increases [Ca2+]i many fold. Use of CaCl2, EGTA and Mn2+ confirmed that both release of Ca2+ from internal pool(s) and influx from outside milieu are responsible for this increase in [Ca2+]i. Application of oligomycin, CN-, and nigericin indicated the non mitochondrial pHi sensitive pool as the possible intracellular origin for internal Ca2+ release. Exposure of these cells to acidic environment had no influence on [Ca2+]i. In contrast, the avirulent promastigotes or freshly isolated amastigotes had shown no effect of heat-shock or pH shock on [Ca2+]i of these cells. Our results indicate that the upward shift in temperature may play a role in signal transduction events in morphogenetic transformation of L. donovani promastigotes that involves mobilization of Ca2+ in cytoplasm. PMID- 7498485 TI - Molecular cloning of a human polypeptide related to yeast sds22, a regulator of protein phosphatase-1. AB - sds22 is a regulatory polypeptide of protein phosphatase-1 that is required for the completion of mitosis in both fission and budding yeast. We report here the cDNA cloning of a human polypeptide that is 46% identical to yeast sds22. The human homolog of sds22 consists of 360 residues, has a calculated molecular mass of 41.6 kDa and shows a tandem array of 11 leucine-rich repeat structures of 22 residues. Northern analysis revealed a major transcript of 1.39 kb in all 8 investigated human tissues. sds22 was detected by western analysis in both the cytoplasm and the nucleus of rat liver cells as a polypeptide of 44 kDa. PMID- 7498486 TI - Ligands of purinergic receptors stimulate electrogenic H(+)-transport of neutrophils. AB - The possible role of ATP, acting as a ligand on cell surface receptor was investigated in the activation of the electrogenic H(+)-transporting pathway of porcine neutrophil granulocytes. (1) ATP brought about 2.1-fold increase in the rate of H(+)-efflux. (2) The order of potency of different nucleotides suggests, that ATP acts on P2 type purinoceptor. (3) The effect of the nucleotides was prevented by inhibition of phospholipase A2. (4) Inhibition of the metabolism of arachidonic acid (AA) via the cyclooxygenase pathway had no effect, whereas inhibition of the lipoxygenase pathway significantly enhanced H(+)-release. This is the first report about activation of the H(+)-transporter by physiological stimulator acting on the cell surface. PMID- 7498488 TI - Stress-induced tyrosine phosphorylation of actin in Dictyostelium cells and localization of the phosphorylation site to tyrosine-53 adjacent to the DNase I binding loop. AB - Actin is known to be phosphorylated at tyrosine, serine, or threonine residues in various cells. In cells of Dictyostelium discoideum, a rise in the tyrosine phosphorylation of actin is observed in response to ATP depletion. An actin fraction rich in phosphotyrosine was obtained by chromatography on the weak anion exchanger Mono-P. Mass spectrometry and amino acid sequencing of protease cleavage products indicated that a single tyrosine residue was phosphorylated. Localization of this residue to position 53 of the actin sequence attributed the modification to a site that is critical for the capability of actin to polymerize. Induction of the tyrosine phosphorylation by heat shock and Cd2+ ions indicates that this modification of actin is implicated in the response of Dictyostelium cells to stress. PMID- 7498489 TI - Direct binding of Torpedo syntrophin to dystrophin and the 87 kDa dystrophin homologue. AB - Syntrophin, a 58-kDa membrane-associated protein, is one component of a protein complex associated with dystrophin and other members of the dystrophin family, including the 87-kDa homologue (87K protein). To characterize interactions between synthrophin and 87K protein, we used an in vitro overlay binding assay. We demonstrate that purified Torpedo syntrophin binds directly to dystrophin and 87K. By expressing overlapping regions of the 87K protein as bacterial fusion proteins for binding targets, we show that a 52-amino acid region of 87K (residues 375-426) is sufficient for binding syntrophin. PMID- 7498490 TI - Site-directed mutagenesis of Lys600 in phosphoenolpyruvate carboxylase of Flaveria trinervia: its roles in catalytic and regulatory functions. AB - Phosphoenolpyruvate carboxylases from various organisms contain two conserved lysine residues. In the C4 dicot Flaveria trinervia, one of these residues is Lys600. Converting this Lys600 to Arg600 or Thr600 mainly increased the Km values and but had minimal effect on the Vmax. The Km for PEP, Mg2+ increased by up to 3 fold in Arg600 and Thr600 but the Km (HCO3-) increased 9-fold in Thr600, suggesting that Lys600 might be associated with bicarbonate-binding. This lysine was not obligatory for enzyme activity although the wild-type protein showed higher activity at physiological pH and was less inhibited by malate than the two mutants. PMID- 7498492 TI - Role of multiple cellular proteases in the execution of programmed cell death. AB - A family of mammalian homologues of the Caenorhabditis elegans cell death protein Ced-3 has been recently discovered. These mammalian proteins encode novel cysteine proteases with homology to the interleukin-1 beta converting enzyme (ICE). Although several studies support a role for one or more of these proteases in mediating apoptosis, their mechanism of action is far from understood. The presence of multiple mammalian ICE-like proteases, with apparently similar apoptotic function indicates that, despite its conservation during evolution, the cell death pathway is much more complex in mammals than in the worm. In addition to ICE-like proteases, several other proteases of different cleavage specificities have been implicated in apoptosis. There is now a growing body of evidence suggesting that apoptosis involves the activation of a cascade of proteases. This article summarises the presently available evidence and discusses how multiple proteases might be required in the effector phase of cell death. PMID- 7498493 TI - Aggregated DNA in ethanol solution. AB - A recently developed mechanochemical method has provided a new, efficient tool for studies on the thermal stability and structure of aggregated DNA in ethanol water solutions. At low ethanol concentrations DNA is fully soluble and is in the B form. However, with increasing ethanol concentration the melting temperature of DNA, Tm, decreases. At a critical ethanol concentration, dependent on the nature and concentration of the counterion, aggregation of the DNA molecules sets in. This is reflected in a marked increase in Tm indicating that the aggregated DNA molecules are thermally more stable than the dissolved ones. However, they are still in the B form. In general, Tm of aggregated DNA also decreases with further increasing ethanol concentration and is dependent on the nature of the counterion, but Tm is not affected by the concentration of the counterion (excess salt) in the ethanol-water solution. When the ethanol concentration reaches the range of 70-80% (v/v), the B-to-A conformational transition occurs in the case of Na-, K- and CsDNA. Above this transition point the A form is more stable than the B form due to the reduced water activity and to increased interhelical interactions. At very high ethanol concentrations, above 85% and dependent on the nature of the counterion, a drastic change in the thermal behaviour is observed. Apparently such a strong interhelical interaction is induced in the aggregated DNA that the DNA is stabilized and cannot adopt a random coil state even at very high temperatures. This stability of DNA in the P form is fully reversed if the ethanol concentration is lowered and the activity of water, thereby, is restored. PMID- 7498491 TI - The recombinant GABA transporter GAT1 is downregulated upon activation of protein kinase C. AB - Treatment of human embryonic kidney 293 cells expressing the rat gamma aminobutyric acid (GABA) transporter 1 (GAT1) with the protein kinase C (PKC) activator phorbol 12-myristate 13-acetate (PMA) was found to decrease the velocity of specific [3H]GABA uptake. This downregulation varied with extracellular GABA concentration and was blocked by the PKC inhibitors 1-(5 isoquinolinylsulphonyl)-2-methylpiperazine (H7) and staurosporine. An about 50% reduction of uptake velocity by PMA treatment was observed at GABA concentrations > 1 microM, whereas only a minor effect was seen at low substrate concentrations. These data indicate that GAT1 activity is downregulated by PKC activation. PMID- 7498494 TI - DNA damage in human respiratory tract epithelial cells: damage by gas phase cigarette smoke apparently involves attack by reactive nitrogen species in addition to oxygen radicals. AB - Treatment of human respiratory tract tracheobronchial epithelial cells with gas phase cigarette smoke led to dose-dependent DNA strand breakage that was highly correlated with multiple chemical modifications of all four DNA bases. The pattern of base damage suggests attack by hydroxyl radicals (OH.). However, by far the most important base damage in quantitative terms was formation of xanthine and hypoxanthine, presumably resulting from deamination of guanine and adenine respectively. Hence, DNA damage by cigarette smoke may involve reactive nitrogen species as well as reactive oxygen species. PMID- 7498495 TI - Co-expression of the neurokinin NK2 receptor and G-protein components in the fission yeast Schizosaccharomyces pombe. AB - The fission yeast Schizosaccharomyces pombe has proven useful for studying molecular interactions between a range of signal transduction components. We now report the first co-expression of a mammalian seven-transmembrane receptor and G protein components in S. pombe. We selected the human neurokinin NK2 receptor together with its G-protein-signalling partner Gq for this study. Yeast membrane fractions showed high levels of NK2 receptor-binding activity (1159 +/- 534 (n = 3) fmol/mg protein) although initial experiments with intact cells revealed an absence of receptors at the cell surface. Using a construct comprising the NK2 coding sequence fused with the signal sequence from an endogenous phosphatase (phoI), we detected approximately 400 NK2 receptors/cell in unbroken yeast. Successful co-expression of the NK2 receptor with the G-protein subunits G alpha q, beta 1 or beta 2 and gamma 3 failed to modulate agonist binding, suggesting the absence of functional interaction between these components. As an alternative test of G alpha q function, we next expressed its downstream effector target phospholipase C-beta 1 (PLC beta 1) in S. pombe. Although PLC beta 1 undergoes powerful in vitro activation by G alpha q derived from baculovirus-infected Sf9 cells and mammalian cells, G alpha q expressed in S. pombe is totally ineffective. Similar results were also achieved with the G-protein subunit G alpha 16. Together, these data suggest that seven-transmembrane receptors can be expressed in S. pombe at high levels and directed to the cell surface although their interaction with co-expressed G-proteins in undetectable. Production of inactive G alpha-chains in S. pombe may account for these observations. PMID- 7498497 TI - Subunit-specific inhibition of inward-rectifier K+ channels by quinidine. AB - Distinct inward-rectifier K+ channel subunits were expressed in Xenopus oocytes and tested for their sensitivity to the channel blocker quinidine. The 'strong' inward-rectifier K+ channel IRK1 was inhibited by quinidine with an EC50 of 0.7 mM, while the 'weak' rectifier channel ROMK1 was only moderately inhibited. ROMK1(N171D)-IRK1C-term chimeric channels, which carry both sites for strong rectification of IRK1 channels (the negatively charged D171 in the second transmembrane domain and the IRK1-C-terminus including E224), displayed strong rectification like IRK1, but showed weak sensitivity to quinidine-like ROMK1, suggesting independence of quinidine binding and rectification mechanisms. Moreover, BIR10 and BIR11, two strong rectifier subunits originally cloned from rat brain, exerted subunit-specific sensitivity to quinidine, being much higher for BIR11. Quinidine blockade of IRK1 was not voltage-dependent, but strongly dependent on the pH in the superfusate. These results strongly suggest a subunit specific interaction of inward-rectifier K+ channels with neutral quinidine within membrane lipid bilayers. PMID- 7498496 TI - Photolabeling acyl CoA binding proteins in microsome preparations. AB - To identify key enzymes that participate in acylglycerol metabolism, 12-[(5-iodo 4-azidosalicyl)amino]dodecanoyl-coenzyme A (ASACoA) was employed as a photoaffinity label for those enzymes that use fatty acyl CoA as a co-substrate. ASACoA inhibited diacylglycerol acyltransferase activity in liver microsomes and was incorporated into triacylglycerol in a microsome dependent reaction. When photoactivated, ASACoA labeled four proteins in rat liver (75, 54, 50 and 37 kDa) and in epididymal fat cell (75, 64, 54 and 37 kDa) microsomes. Photolabeling was sensitive to palmitoyl CoA. After solubilization in Triton X-114, all four proteins were concentrated into the detergent phase, indicating that they are integral membrane proteins. PMID- 7498498 TI - HiPiP oxido-reductase activity in membranes from aerobically grown cells of the facultative phototroph Rhodoferax fermentans. AB - The role of the periplasmically located, water-soluble, HiPIP (high-potential iron-sulfur protein) in the respiratory chain of the facultative phototroph Rhodoferax fermentans has been examined. The oxidized HiPIP is reduced by succinate-dependent respiration via the bc1 complex, this reaction being inhibited by myxothiazol and/or stigmatellin. The reduced HiPIP can be oxidized by the membrane-bound cytochrome oxidase, this reaction being inhibited by 0.1 mM cyanide. We conclude that aerobically grown Rf. fermentans contains a redox chain in which HiPIP mediates electron transfer between the bc1 complex and the cb-type cytochrome oxidase. PMID- 7498500 TI - Nature of respiratory stimulation in hyperthyroidism: the redox behaviour of cytochrome c. AB - Hyperthyroid mitochondria show an increased Km and Vmax in the high affinity phase of cytochrome oxidase kinetics. During inhibitor titrations, cytochrome c shows a different redox behaviour in hyperthyroid with respect to protonophore treated euthyroid mitochondria. The observations are discussed in terms of a different regulation of electron input and output into the respiratory chain during slip and leak types of uncoupling. In hyperthyroid mitochondria during inhibitor titrations, the pattern of the relationship between uncoupler-induced extra-respiration and membrane potential is highly non-linear. The complex nature of the respiratory stimulation in hyperthyroid mitochondria is discussed. PMID- 7498499 TI - Evidence for a metabostatic opioid kappa-receptor inhibiting pertussis toxin sensitive metabotropic glutamate receptor-currents in Xenopus oocytes. AB - Glutamate evoked pertussis toxin-sensitive currents in Xenopus oocytes expressing metabotropic glutamate receptor subtype 1 (mGluR1) and exogenous Gi1 alpha. The mGluR1-currents were completely blocked by U-73122, a phospholipase C (PLC) inhibitor and by niflumic acid, a chloride channel blocker. In the oocyte further coinjected with poly(A)+ RNA from the guinea pig cerebellum, the mGluR1-currents were inhibited by U-50488H, an opioid kappa-agonist, and this inhibition was blocked by norbinaltorphimine, an opioid kappa-antagonist. These findings suggest that the mRNA encoding a novel subtype of opioid kappa-receptor which inhibits Gi1-PLC-mediated currents is present in guinea pig cerebellar poly(A)+ fractions. PMID- 7498502 TI - Dose-dependent activation and block by bisG10, a K+ channel blocker, of mouse and frog skeletal muscle KATP channels. AB - The effects of a K+ channel blocker, bisG10, were examined on ATP-sensitive K+ (KATP) channels in membrane patches excised from mammalian and amphibian skeletal muscle fibres using the patch-clamp technique. At micromolar concentrations, bisG10, added on the intracellular side, induced a strong, reversible, flickery block of KATP channels. BisG10, added on the extracellular side, was about 100 fold less potent at inhibiting channel activity. At 10 nM, intracellular bisG10 increased KATP channel activity. This activation was independent of the presence of internal ATP or Mg2+. The inhibitory effect of bisG10 most likely arose from open-channel block whereas activation could result from more complex, indirect interactions. PMID- 7498501 TI - Synthesis of chloramphenicol acetyltransferase in a coupled transcription translation in vitro system lacking the chaperones DnaK and DnaJ. AB - A trimeric enzyme chloramphenicol acetyltransferase (CATI) has been synthesized in the Zubay system genetically depleted from DnaK and DnaJ. Most of CAT formed in the system fail to assemble into an active trimer. Instead CAT is accumulated in either a GroEL-bound complex or as an inactive monomer. Addition of purified DnaK and DnaJ to the system prior to the start of protein synthesis leads to the increase of the specific activity of formed CAT. A portion of exogenous DnaK and DnaJ added to the system associate with nascent polypeptide chains in the ribosomes. DnaK also comigrates with 50S-ribosomal subunits. PMID- 7498503 TI - The presence of a hydroxyl group at the C-1 atom of the transketolase substrate molecule is necessary for the enzyme to perform the transferase reaction. AB - Transketolase catalyzes the transfer of an aldehyde residue from keto sugars to aldo sugars. The intermediate product is dihydroxyethylthiamine pyrophosphate (DHETPP). In the absence of an acceptor substrate, the reaction is stopped at this stage and DHETPP does not undergo subsequent transformations. Pyruvate decarboxylase catalyses pyruvate decarboxylation to yield free aldehyde. The intermediate product is hydroxyethylthiamine pyrophosphate (HETPP). It differs from DHETPP only in that it has no hydroxyl at the C-2 atom of the aldehyde residue. We have shown that transketolase can bind HETPP and split the aldehyde residue from it. This fact suggests that the path of the reaction is determined by the absence (in HETPP) or presence (in DHETPP) of a hydroxyl group. In the former case the reaction will yield free aldehyde, in the latter the aldehyde residue will be transferred onto an acceptor substrate. PMID- 7498504 TI - High-specific binding of Fe(II) at the Mn-binding site in Mn-depleted PSII membranes from spinach. AB - The interaction of Fe(II) and Fe(III) with the 'high-affinity Mn-binding site' in Mn-depleted photosystem II (PSII) was investigated using diphenilcarbazide (DPC)/2,6-dichlorophenol-indophenol (DCIP) inhibition assay. Fe(III) was ineffective in the inhibition of DPC-DCIP reaction while Fe(II) decreased the rate of DCIP photoreduction supported by DPC in the same concentration range as Mn(II). The effectivity of the interaction of Fe(II) with the high affinity Mn binding site depends on different anions in the same manner as for Mn(II) and coincides with hierarchy observed for the stimulation of O2 evolution. The Fe(II) binding is accompanied by its oxidation. By using reductants it was shown that the high affinity site contains a redox-active component and the reduction of this component totally prevents the binding of Fe(II). PMID- 7498505 TI - Plasma membrane fatty acid-binding protein (FABPpm) is exclusively located in the maternal facing membranes of the human placenta. AB - We reported earlier the presence of a 40 kDa plasma membrane fatty acid-binding protein (FABPpm) in human placenta. This protein is thought to be involved in the sequestration of unesterified free fatty acids bound to albumin from the maternal plasma for delivery to the fetus. However, its location in human placental syncytiotrophoblasts is not known. These cells are bipolar; one side facing maternal circulation (microvillous membranes), and the other side facing fetal circulation (basal membranes). Therefore, it is important to resolve the location of this protein in trophoblast membranes in order to understand fatty acid transport and metabolism in human placenta. Isolated plasma membranes vesicles were prepared respectively from the maternal facing microvillous and fetal facing surface of the human full-term placental syncytiotrophoblast. Using these membrane preparations, fatty acid binding activity, the polyacrylamide gel electrophoresis radiobinding assay for FABPpm, and Western blot analysis of FABPpm were carried out to determine the location of this protein in these membranes. Based on the above studies we conclude that the FABPpm is located exclusively in the microvillous membranes. Since FABPpm may be responsible for FFA uptake, its location in the microvillous membranes favours the unidirectional flow of maternal FFA to the fetus. PMID- 7498506 TI - (RP)-cAMPS inhibits the cAMP-dependent protein kinase by blocking the cAMP induced conformational transition. AB - (RP)-cAMPS is known to inhibit competitively the cAMP-induced activation of cAMP dependent protein kinase (PKA). The molecular nature of this inhibition, however, is unknown. By monitoring the intrinsic tryptophan fluorescence of recombinant type I regulatory subunit of PKA under unfolding conditions, a free energy value (delta GDH2O) of 8.23 +/- 0.22 kcal/mol was calculated. The cAMP-free form of the regulatory subunit was less stable with delta GDH2O = 6.04 +/- 0.05 kcal/mol. Native stability was recovered by treatment of the cAMP-free protein with either cAMP or (SP)-cAMPS but not with (RP)-cAMPS. Thus, (RP)-cAMPS binding to the regulatory subunit keeps the protein in a locked conformation, unable to release the catalytic subunit. This finding was further supported by demonstrating that holoenzyme formation was greatly accelerated only when bound cAMP was replaced with (RP)-cAMPS but not with cAMP or (SP)-cAMPS. PMID- 7498507 TI - Expression, purification and kinetic behaviour of fission yeast low M(r) protein tyrosine phosphatase. AB - A gene named stp1+, coding for a 17.5-kDa protein, that rescues cdc25-22 when overexpressed, has been previously isolated from fission yeast. Here we describe the expression and purification of Stp1 protein as a fusion with the glutathione S-transferase in E. coli and its kinetic characterisation. Stp1 deduced protein sequence shows an high homology to members of a class of cytosolic low M(r) protein phosphatase previously known to exist only in mammalian species. Stp1 has a kinetic behaviour that appears to be intermediate with respect to the two isoenzymatic forms of low M(r) protein tyrosine phosphatases present in mammalian tissues. These differing kinetic characteristics are mainly due to the sequence 45-56 that is spatially close to the active site pocket. PMID- 7498508 TI - Conformational changes upon binding of a receptor loop to lipid structures: possible role in signal transduction. AB - The mas oncogene codes for a seven transmembrane helix protein. The amino acid sequence 253-266, from the third extracellular loop and beginning of helix 7, was synthesized either blocked or carrying an amino acid spin label at the N terminus. Peptide binding to bilayers and micelles was monitored by ESR, fluorescence and circular dichroism. Binding induced tighter lipid packing, and caused an increase of peptide secondary structure. While binding to bilayers occurred only when peptide and phospholipid bore opposite charges, in micelles the interaction took place irrespective of charge. The results suggest that changes in lipid packing could modulate conformational changes in receptor loops related to the triggering of signal transduction. PMID- 7498509 TI - Tau isoform expression and phosphorylation state during differentiation of cultured neuronal cells. AB - The axonal microtubule-associated protein, tau, is thought to play an important role in axonal growth and in the establishment of neuronal polarity. In adult human brain there are six alternatively spliced tau isoforms, which have different microtubule binding affinities in vitro. The tubulin-tau interaction is further modified by phosphorylation of tau and, compared to adult brain tau, both foetal brain tau and paired helical filament (PHF) tau, characteristic of Alzheimer's disease, are hyperphosphorylated. In vivo both the expression of tau isoforms and their phosphorylation states are developmentally regulated. In order to establish the correlation between the expression of tau isoforms and their pattern of phosphorylation, we have characterised these two features in several in vitro models of neuronal differentiation, including the human neuroblastoma cell lines, SK-N-SH, SH-SY5Y and IMR32 cells, rat PC12 cells and primary rat cortical neurones. Sensitive RT-PCR analysis revealed a different complement of tau isoforms in the different cell lines and neuritogenesis was associated mainly with an increase in the overall tau protein level with no apparent phosphorylation changes. A switch in tau isoform expression occurred only at the terminal stages of neuronal development, when it may be important in reinforcing the previously established axonal cytoarchitecture. PMID- 7498510 TI - Inhibition of glycosphingolipid synthesis induces p34cdc2 activation in Xenopus oocyte. AB - In Xenopus prophase-blocked oocytes, it is assumed that progesterone interacts with the plasma membrane to initiate a signalling cascade that ultimately leads to MPF activation. Progesterone regulates negatively the cAMP pathway through an inhibition of adenylate cyclase. However, the mechanisms linking the initial action of the hormone with adenylate cyclase activity remain to be elucidated. Here, we demonstrate that PDMP, an inhibitor of glucosphingolipid synthesis, triggers oocyte meiotic maturation in a cAMP- and cycloheximide-dependent manner, whereas exogenous ceramide is unefficient. We propose that sphingolipid metabolism and targeting represent an important regulatory process of oocyte meiosis. PMID- 7498511 TI - Mycobacterial glycopeptidolipid interactions with membranes: a monolayer study. AB - Mycobacterial glycopeptidolipid (GPL) interactions with membranes were analysed with monolayer experiment, using GPLs bearing 3, 1, or 0 carbohydrate residues (GPL3, GPL1, GPL0). Compression isotherms and surface potential determinations suggested that the glycopeptidic moiety of GPL3 permanently dipped in water, while those of GPL1 and GPL0 can lay in the interface. Insertion of GPL molecules into a preformed phospholipid monolayer was observed using GPL3 or GPL1 dispersions, but not from GPL0. It is postulated that the activity of GPL0 is low due to its failure to become inserted into membranes, as is that of GPL3 owing to its insertion only by its acyl chain. GPL1 is likely to disturb membranes by inserting its glycopeptidic moiety into the interface. PMID- 7498512 TI - Structural analysis of recombinant von Willebrand factor produced at industrial scale fermentation of transformed CHO cells co-expressing recombinant furin. AB - Thorough analysis of multimer composition and molecular structure of recombinant von Willebrand factor (r-vWF) produced by recombinant CHO cells demonstrated r vWF to be more intact and less proteolytically degraded than plasma-derived vWF (pd-vWF) [B. Fischer et al. (1994) FEBS Lett. 351, 345-348]. In contrast to pd vWF, r-vWF preparations consisted of pro-vWF (vWF containing covalently attached propeptide) as well as mature vWF subunits forming homo- and hetero-multimers. In order to ensure complete propeptide processing, a r-vWF-producing CHO cell clone was transfected with the cDNA of the human propeptide processing enzyme Furin. A r-vWF/r-Furin co-expressing cell clone was cultivated at industrial scale in high cell density perfusion fermenters. r-vWF obtained from these cells was fully processed. Analysis of r-vWF by multimer analysis revealed a multimer pattern equal in number of high molecular weight multimer to pd-vWF, but absence of satellite bands. Two-dimensional electrophoretic analysis of both the primary dimer and the complete multimer pattern of r-vWF showed that the recombinant coagulation factor was composed exclusively of intact and mature subunits. Since the triplet structure typical to pd-vWF is known to reflect proteolytic degradation, r-vWF thus exhibits an integrity far superior compared to pd-vWF. PMID- 7498513 TI - Expression of the human cGMP-dependent protein kinase II gene is lost upon introduction of SV40 T antigen or immortalization in human cells. AB - We have cloned a human cGMP-dependent protein kinase type II cDNA to examine its gene expression in terms of cellular senescence and/or immortalization. The genetic locus was mapped to band 4q21 by FISH. Northern blot analysis revealed that expression of the type II gene was markedly decreased or lost in mortal or immortal human fibroblasts producing SV40 T antigen. Also in various immortalized cell lines tested, the gene was not expressed. In normal diploid fibroblasts, the gene was constitutively expressed during cell-cycle and population doubling levels (PDLs). PMID- 7498514 TI - Down-regulation of mitochondrial mRNAs in the mdx mouse model for Duchenne muscular dystrophy. AB - In our search for genes up- or down-regulated genes in the mdx mouse model for Duchenne muscular dystrophy, we isolated a down-regulated mitochondrial DNA clone. In addition to this clone, all protein-coding mitochondrial genes tested had tissue-specific and age independent down-regulated expression. This implied mechanisms at the RNA level since no change in the mitochondrial DNA contents were detected. Cytochrome c oxidase activity showed the same range of down regulated expression. These data provide a molecular basis for energetic metabolism modifications in mdx mice. PMID- 7498517 TI - Purification from pig kidney of a microsomal cytochrome P450 catalyzing 1 alpha hydroxylation of 25-hydroxyvitamin D3. AB - A cytochrome P450 catalyzing 1 alpha-hydroxylation of 25-hydroxyvitamin D3 was purified from pig kidney microsomes. The enzyme preparation showed one protein band on gel electrophoresis with apparent M(r) of 52,500 and a specific cytochrome P450 content of 10.7 nmol/mg of protein. The 25-hydroxyvitamin D3 1 alpha-hydroxylase copurified with the vitamin D3 25-hydroxylase during purification. A cytochrome P450 catalyzing 1 alpha-hydroxylation was purified also from liver microsomes. The apparently homogeneous enzyme showed the same catalytic properties and apparent M(r) as the kidney enzyme. The results of the present communication demonstrate the presence in kidney of a previously unknown microsomal 1 alpha-hydroxylase in addition to the assumed specific mitochondrial 1 alpha-hydroxylase. The possibility that microsomal 1 alpha-hydroxylation in pig kidney and liver is catalyzed by the previously described porcine microsomal vitamin D 25-hydroxylase(s) is discussed. PMID- 7498515 TI - Phospholipid membrane permeability of peptide nucleic acid. AB - Phospholipid vesicles (liposomes) as membrane models have been used to study the penetration properties of peptide nucleic acid (PNA), a new DNA analog in which the nucleobases are attached to a pseudo-peptide backbone. The liposomes were characterised by carboxyfluorescein efflux, light-scattering and cryogenic transmission electron microscopy. The liposome structure was found not to be affected by the incorporation of PNA or an oligonucleotide. Two 10-mer fluorescein-labelled PNAs were found to have low efflux rates (half-times of 5.5 and 11 days), comparable to that of a 10-mer oligonucleotide (half-time of 7 days). We conclude that passive diffusion of unmodified PNA over the lipid membrane is not likely to be an effective way of transport into biological cells. PMID- 7498518 TI - Enzymatic and immunological activity of 4000 years aged bone alkaline phosphatase. AB - Structurally intact and functionally active human bone alkaline phosphatase was isolated from clavicle fragments of IDU, an Egyptian mummy of the Old Kingdom (2150 +/- 50 BC). Both anion exchange and affinity chromatographies were employed to optimise the preparation of the ancient enzyme resulting in a specific activity of 180 +/- 30 mU/mg. The intactness of the bone enzyme fractions of the wheat-germ lectin affinity chromatography was successfully demonstrated in an ELISA using the monoclonal antibody BAP A. Fortunately, the mummified bone was not contaminated by fungi or bacteria. PMID- 7498516 TI - Thermostable peroxidase activity with a recombinant antibody L chain-porphyrin Fe(III) complex. AB - In order to engineer a new type of catalytic antibody, we attempt to use a monoclonal antibody L chain as a host protein for a porphyrin. TCPP (meso tetrakis(4-carboxyphenyl)porphyine) was chemically synthesized and Balb/c mice were immunized using TCPP as a hapten. Two hybridoma cells (03-1, 13-1), that produce monoclonal antibody against TCPP, were obtained. Genes for both H and L chains of monoclonal antibodies were cloned, sequenced and overexpressed using E. coli as a host. ELISA and fluorescence quenching method show that the independent antibody L chains from both Mab03-1 and Mab13-1 have specific interaction with TCPP. Furthermore, the recombinant antibody L chain from Mab13-1 exhibits much higher peroxidase activity than TCPP Fe(III) alone. The enzyme activity was detectable with pyrogallol and ABTS (2,2-azinobis-3-ethylbenzthiazolin-6-sulfonic acid) but not with catechol. This new catalytic antibody was extremely thermostable. Optimum temperature of the peroxidase reaction by the complex of 13 1L chain and TCPP Fe(III) was 90 degrees C, while that the TCPP Fe(III) alone was 60 degrees C. PMID- 7498519 TI - An interleukin-1 beta-converting enzyme-like protease is a common mediator of apoptosis in thymocytes. AB - Apoptosis was induced in thymocytes using diverse stimuli in order to identify events within a common apoptotic pathway. Benzyloxycarbonyl-valinyl-alaninyl aspartyl fluoromethyl ketone (Z-VAD.FMK), an interleukin-1 beta-converting enzyme (ICE)-like protease inhibitor, inhibited apoptosis assessed by flow cytometry, proteolysis of poly (ADP)-ribose polymerase of DNA to both large kilobase pair fragments (30-50 and 200-300 kbp) and to nucleosomal fragments. Z-VAD.FMK also blocked all the classical ultrastructural features of apoptosis including chromatin condensation to one pole of the nucleus, nucleolar disintegration and cytoplasmic vacuolation. These results suggest the involvement of an ICE-like protease as a common mediator of apoptosis in thymocytes. PMID- 7498520 TI - Comparison of the specificities of p70 S6 kinase and MAPKAP kinase-1 identifies a relatively specific substrate for p70 S6 kinase: the N-terminal kinase domain of MAPKAP kinase-1 is essential for peptide phosphorylation. AB - xxR/KxRxxSxx sequences were phosphorylated with high efficiency by both p70 S6 kinase (p70S6K) and MAPKAP kinase-1. The best substrate for MAPKAP kinase-1 (KKKNRTLSVA) was phosphorylated with a Km of 0.17 microM, and the best substrate for p70S6K (KKRNRTLSVA) with a Km of 1.5 microM. The requirement of both enzymes for Arg/Lys at position n-5 could be partially replaced by inserting basic residues at other positions, especially by an Arg at n-2 or n-4. MAPKAP kinase-1 (but not p70S6K) tolerated lack of any residue at n-5 if Arg was present at n-2 and n-3. p70S6K (but not p90S6K) tolerated Thr at position n and absence of any residue at n + 2. The peptide KKRNRTLTV, which combined these features, was relatively selective for p70S6K having a 50-fold higher Vmax/Km than MAPKAP kinase-1. Inactivation of the N-terminal kinase domain of MAPKAP kinase-1, which is 60% identical to p70S6K, abolished activity towards all peptides tested, but the enzyme retained 30-40% of its activity if the C-terminal kinase domain was inactivated. PMID- 7498521 TI - Amino acid sequence and expression of the hepatic glycogen-binding (GL)-subunit of protein phosphatase-1. AB - A full-length cDNA encoding the putative hepatic glycogen-binding (GL) subunit of protein phosphatase-1 (PP1) was isolated from a rat liver library. The deduced amino acid sequence (284 residues, 32.6 kDa) was 23% identical (39% similar) to the N-terminal region of the glycogen-binding (GM) subunit of PP1 from striated muscle. The similarities between GM and GL were most striking between residues 63 86, 144-166 and 186-227 of human GM (approximately 40% identity), nearly all the identities with the putative yeast homologue GAC1 being located between 144-166 and 186-227. The cDNA was expressed in E. coli, and the expressed protein transformed the properties of PP1 to those characteristic of the hepatic glycogen associated enzyme. These experiments establish that the cloned protein is GL. PMID- 7498522 TI - Up-regulation of protein serine/threonine phosphatase type 2C during 1 alpha,25 dihydroxyvitamin D3-induced monocytic differentiation of leukemic HL-60 cells. AB - Treatment with 20 nM 1 alpha,25-dihydroxyvitamin D3 (1,25(OH)2D3) caused a progressive increase in the activity of Mg(2+)-dependent protein serine/threonine phosphatase type 2C (PP2C) in subcellular fractions of HL-60 cells, whereas PP2C activity was relatively constant throughout all-trans retinoic acid-induced (1 microM) granulocytic differentiation. The increase in PP2C activity appeared to parallel the 1,25(OH)2D3-induced phenotypic and functional changes in HL-60 cells. Immunoblot and Northern blot analysis indicated that the increase in PP2C activity corresponded to the increased expression of PP2C protein, which was preceded by an increase in the level of mRNA for PP2C beta. No mRNA for PP2C alpha was detected in resting or 1,25(OH)2D3-stimulated HL-60 cells. These results suggest that the increased expression of PP2C is related with the 1,25(OH)2D3-induced monocytic differentiation of HL-60 cells. PMID- 7498523 TI - Mono-, di- and trimeric binding of a bis-netropsin to DNA. AB - An unusual 3:1 stoichiometry for complex formation between an elongated bis netropsin compound and its binding site on DNA has been observed. Circular dichroism measurements distinguish two types of complexes formed between this bis netropsin and poly[d(A-T)].poly[d(A-T)]. The first type is characterized by a 1:1 saturating ratio of bound molecules per ten base pairs. Formation of the second type results from the cooperative binding of two additional bis-netropsin molecules to the first type of complex. In contrast to these results observed for binding to the alternating polynucleotide, only the 1:1 type of complex is formed when this ligand binds to the homopolymer poly(dA).poly(dT). PMID- 7498524 TI - A human homolog of the mitochondrial protein import receptor Mom19 can assemble with the yeast mitochondrial receptor complex. AB - Import of preproteins into mitochondria requires transport machineries in both mitochondrial membranes that have been characterized in Saccharomyces cerevisiae and Neurospora crassa. By cDNA analysis, we identified a human protein of 16 kDa with significant overall homology to the fungal mitochondrial import receptor Mom19, including the three typical characteristics: a hydrophobic N-terminal segment, a tetratrico peptide motif in the middle and a negatively charged C terminus. The human Mom19 homolog is expressed in all tissues analyzed. When synthesized in vitro, the human Mom19 homolog is targeted to isolated yeast mitochondria and specifically associates with the outer membrane receptor complex, suggesting that indeed a mitochondrial import receptor was identified. PMID- 7498525 TI - Analysis of CpG dinucleotide frequency in relationship to translational reading frame suggests a class of genes in which mutation of this dinucleotide is asymmetric with respect to DNA strand. AB - Results are described from application of a computer program that compares the expected and actual incidence of CpG dinucleotides in relation to the codon reading frame of genes, assuming a conserved amino acid sequence and normalizing for the third-position incidences of C and G in the remainder of the sequence. Sequences encoding certain proteins showed a pronounced bias in favour of CpG in the (3,1) compared with the (2,3) codon position; whereas sequences encoding related proteins expressed to a similar extent or in the same tissue did not. We propose that the cases exhibiting this bias reflect a difference between the two strands of the DNA duplex in their susceptibility to loss of CpG dinucleotides by mutation. Although in vertebrates this loss of CpG dinucleotides from the sense strand might reflect strand-asymmetry in deamination of 5-methylcytosine residues, the fact that a similar CpG codon bias is found in some invertebrates indicates that other factor(s) must also be involved. PMID- 7498526 TI - Structures of genes for two cathelin-associated antimicrobial peptides: prophenin 2 and PR-39. AB - We characterized genes for prophenin (PF)-2 and PR-39, two cathelin-associated antimicrobial peptides found in porcine leukocytes. Both contained 4 exons and 3 introns and were compact, contiguous and highly homologous. Exons I-III encoded most of their cathelin domains. Exon IV specified the final few cathelin residues, including its conserved C-terminal valine, followed by the mature PR-39 peptide or a PF-2 precursor. The highly conserved 5' flanking sequences of this gene family contained NF-kappa B, IL-6, GM-CSF and NF-1 binding motifs and the introns were unusually conserved. These data suggest that the panoply of porcine cathelin-associated antimicrobial peptides arose relatively recently via gene reduplications and exon shuffling, and that in vivo expression of cathelin associated antimicrobial peptides may respond to mediators generated early during infection. PMID- 7498527 TI - Characterisation of a synergohymenotropic toxin produced by Staphylococcus intermedius. AB - Staphylococcal synergohymenotropic (SHT) toxins damage membranes of host defence cells and erythrocytes by the synergy of two secreted and non-associated proteins: class S and class F components. Whereas Panton-Valentine leucocidin (PVL), gamma-hemolysin and Luk-M from Staphylococcus aureus are members of this toxin family, a new bi-component toxin (LukS-I + LukF-I) from Staphylococcus intermedius, a pathogen for small animals, was characterised and sequenced. It is encoded as a luk-I operon by two cotranscribed genes, like PVL, LukS-I + LukF-I shares a strong leukotoxicity of various PMNs, but only slight haemolytic properties on rabbit erythrocytes. When intradermally injected into rabbit skin, a 100 ng dose caused acute inflammatory reaction leading to tissue necrosis. The new SHT seemed to be largely distributed among various Staphylococcus intermedius strains. PMID- 7498528 TI - Involvement of phosphatidylinositol 3-kinase in the activation of extracellular signal-regulated kinase by PDGF in hepatic stellate cells. AB - Phosphatidylinositol 3-kinase (PI 3-K) is a lipid and protein kinase which associates with the activated platelet-derived growth factor (PDGF) receptor and other tyrosine kinases. We studied the effects of wortmannin, a selective inhibitor of PI 3-K, on the activation of extracellular-signal regulated kinase (ERK) by PDGF in cultured hepatic stellate cells, mesenchymal cells responsible for extracellular matrix synthesis within the liver. Incubation with 100 nM wortmannin, a dose which almost completely blocks PI 3-K, resulted in 50% reduction of ERK activity. Direct inhibition of ERK by wortmannin could not be considered responsible for this effect, since wortmannin did not inhibit ERK activity in vitro. Rather, inhibition of PI 3-K acts on the kinase cascade that leads to ERK activation, since PDGF-dependent phosphorylation of ERK was found to be reduced after incubation with wortmannin. Wortmannin also inhibited the increase in c-fos mRNA induced by PDGF, which is dependent on ERK activation. The results of this study show that in hepatic stellate cells PI 3-K is involved in ERK activation, although it is not necessary. These data indicate cross-talk between PI 3-K and the Ras/ERK pathway in PDGF-stimulated cells. PMID- 7498529 TI - Unusual long target duplication by insertion of intracisternal A-particle element in radiation-induced acute myeloid leukemia cells in mouse. AB - Retrotransmission into the IL-3/GM-CSF gene locus by the retrotransposon intracisternal A-particle (IAP) had been observed in distinct tumor cell lines. We analyzed the locus in genomes from 7 different myeloid leukemia cell strains which were originally generated by whole-body X-irradiation of the inbred C3H/He mice at a dose of 3 Gy and maintained by in vivo passage. In one leukemia cell strain out of 7 such cases, RFLP of an allele of the interleukin-3 gene was found. Sequence analysis after cloning from the genomic library showed that a type I delta 2 IAP element was inserted in the region upstream of the IL-3 gene in the head-to-head orientation. This suggests that the locus in myeloid cells is sensitive for integration of IAP elements. Additionally, an unusual long target duplication of 82 bp, 14-fold larger than normal one, was found at the junction of the element. This suggests the possibility of a radiation-induced integration mechanism which is distinct from normal retrotransmission. PMID- 7498530 TI - Steroid hormones and temperature induce changes of binding parameters of their receptors in intact cells. AB - When MCF-7 cells were treated with 17 beta-estradiol, dexamethasone, or promegestone at 37 degrees C, the KD of receptors for their cognate ligands was found to decrease as compared to that measured at 2 degrees C. Cell incubation with hormone at 37 degrees C did not affect the Bmax of glucocorticoid and progesterone receptors, but caused a 40% increase of that of estrogen receptor. This increase required the presence of ligand, was insensitive to cycloheximide, and was completed within 10 min of cell incubation at physiological temperature. We conclude that an early step of estrogen action is the recruitment of pre existing receptor molecules through activation of their binding capacity. PMID- 7498532 TI - Influence of extracellular matrices on ganglioside pattern of two hepatoma cell lines with different adhesive properties. AB - A cell culture model was developed to investigate the involvement of gangliosides in cell-matrix adhesion. Two cell lines with different adhesive properties derived from solid Morris hepatoma 7777 were established. Cultured in horse serum containing medium, the adhesive cell line (MH 7777A) adheres and spreads on uncoated culture dishes, whereas the revertant cell line (MH 7777A > N) does not adhere and grows in suspension. The adhesiveness of both cell lines is dependent on the coating protein used (none, bovine serum albumin, fibronectin or collagen I) and the horse serum concentration in the culture medium. Both cell lines, although of the same origin, differed in their ganglioside composition. The most abundant ganglioside of both MH 7777A and MH 7777A > N cell lines was fucosyl GM1, 0.78 and 0.72 microgram per mg cellular protein, respectively. The GM3 and GD1a content of MH 7777A > N cells was significantly higher than that of MH 7777A cells. Furthermore, a matrix-dependency of the ganglioside pattern of both cell lines was demonstrated. PMID- 7498533 TI - The phorbol 12-myristate 13-acetate (PMA)-induced oxidative burst in rat peritoneal neutrophils is increased by a 0.1 mT (60 Hz) magnetic field. AB - Magnetic fields (MF) may affect biological systems by increasing free radical concentrations. To test this, we have investigated whether low frequency (60 Hz) low intensity (0.1 mT) MF can modulate the phorbol 12-myristate 13- acetate (PMA) induced respiratory burst in primed rat peritoneal neutrophils, followed in real time using the dye 2',7'-dichlorofluorescin (DCFH), which reacts with free radical-derived oxidants such as H2O2 (which is formed from the dismutation of superoxide) to become 2',7'-dichlorofluorecein (DCF), a highly fluorescent compound. In the presence of the MF, a 12.4% increase in the fluorescence signal was observed in PMA-stimulated neutrophils (n = 5, P < 0.02, 18 pairs of measurements). We believe this represents the first experimental observation of MF influencing events involving free radical species generated during signal transduction in living cells. PMID- 7498531 TI - Human lymphoblast and erythrocyte multicatalytic proteases: differential peptidase activities and responses to the 11S regulator. AB - The multicatalytic protease (MCP) or 20S proteasome was purified from human red blood cells and two lymphoblastoid cell lines, 721.45 which constitutively expresses protease subunits LMP2 and LMP7, and 721.174 in which genes for these subunits are deleted. Each MCP was assayed using a series of fluorogenic peptides. The hydrophobic peptides gGGF-MCA, sRPFHLLVY-MCA and sLY-MCA were particularly good substrates for 721.45 MCP as compared to the enzyme from 721.174 and red blood cells. In addition, hydrolysis of gGGF-MCA and sLY-MCA was activated by human red blood cell and recombinant regulators to a greater extent using MCP from 721.45 lymphoblasts. Thus, LMP2/LMP7 and regulator appear to act synergistically in the enhanced degradation of gGGF-MCA and sLY-MCA by the multicatalytic protease. PMID- 7498535 TI - Identification of a phosphatidylinositol-4,5-bisphosphate-binding domain in the N terminal region of ezrin. AB - Purified human recombinant ezrin cosediments with large liposomes containing phosphatidylserine (PS). This interaction is optimal at low ionic strength. At physiological ionic strength (130 mM KCl) ezrin interacts strongly with liposomes containing > or = 5% phosphatidylinositol-4,5-bisphosphate (PIP2), the residual being phosphatidylcholine (PC). When PIP2 is replaced by phosphatidylinositol-4 monophosphate (PIP), phosphatidylinositol (PI) or PS, the interaction is markedly reduced. Furthermore we show, that a purified N-terminal glutathione S transferase (GST) fusion protein of ezrin (1-309) still has retained the capacity to interact with PIP2-containing liposomes, whereas a C-terminal fusion protein (310-586) has lost this ability. PMID- 7498534 TI - Evidence for direct involvement of the sarcoplasmic reticulum Ca(2+)-ATPase in a passive monovalent cation (K+/Na+) exchange. AB - A specific inhibitor of SERCA-pumps, thapsigargin (TG) was used to demonstrate the direct involvement of the SR Ca(2+)-ATPase in passive K+/Na+ exchange. The K(+)-potential variations across vesicle membranes were measured in the absence of ATP with a fluorescent probe: 3,3'-dipropylthiodicarbocyanine iodide. Addition of EGTA dissipates the K(+)-potential whereas the presence of TG abolishes this effect. Our data prove that the Ca(2+)-ATPase translocates monovalent cations at a rate similar to the E2-->E1 conformational change. PMID- 7498536 TI - Epidermal growth factor-binding protein activates soluble and receptor-bound single chain urokinase-type plasminogen activator. AB - Epidermal growth factor-binding protein (EGF-BP) is a serine proteinase that reversibly associates with epidermal growth factor (EGF). We analyzed the reaction of EGF-BP with urokinase type plasminogen activator (u-PA), a serine proteinase that promotes pericellular proteolysis and cellular migration. EGF-BP cleaved single chain u-PA (scu-PA) between Lys158 and Ile159, converting the zymogen into enzymatically active two-chain u-PA (tcu-PA), as shown by SDS-PAGE, N-terminal sequence analysis, and enzymatic assay. The kcat and Km of the activation reaction were (5.6 +/- 0.6) x 10(-2)s-1 and 2.0 +/- 0.3 microM, yielding a catalytic efficiency of 2.8 x 10(4) M-1.s-1. EGF-BP also activated scu PA bound to receptors on U937 monocytes as demonstrated by the generation of amidase activity against a tcu-PA-specific fluorogenic substrate. By activating scu-PA, EGF-BP may initiate u-PA-dependent cell surface proteolysis and therefore enhance EGF activities that require cellular migration and/or tissue remodeling. PMID- 7498537 TI - Functional expression of an alpha anti-insect scorpion neurotoxin in insect cells and lepidopterous larvae. AB - The Leiurus quinquestriatus hebraeus alpha anti-insect toxin (Lqh alpha IT) cDNA was engineered into the Autographa californica Nuclear Polyhedrosis Virus (AcNPV) genome. Insect cells infected with the recombinant virus secreted a functional Lqh alpha IT polypeptide. Spodoptera littoralis and Heliothis armigera larvae injected with recombinant budded virus, showed typical intoxication symptoms. This recombinant virus showed enhanced insecticidal potency against H. armigera larvae compared with wild type AcNPV. The present expression system will facilitate: (1) the future elucidation of structural elements involved in its prominent anti-insect toxicity; and (2) the future design of genetically modified alpha toxins with improved anti-insect selectivity. PMID- 7498538 TI - Molecular cloning and characterization of cDNA for insect biogenic peptide, growth-blocking peptide. AB - Growth-blocking peptide (GBP) is an insect biogenic peptide that prevents the onset of metamorphosis from larva to pupa. A cDNA coding for GBP is described. Mixed oligonucleotides derived from a GBP peptide sequence were used to generate amplified DNA by the polymerase chain reaction (PCR). Based on the sequence of the amplified DNA, a 41 bases oligonucleotide was designed for screening a cDNA library which was constructed from the armyworm Pseudaletia separata larvae parasitized with the parasitic wasp Cotesia kariyai. The cloned cDNA for GBP was 809 base pairs in length. An open reading frame of 429 base pairs encodes a pre pro-peptide of 143 amino acid residues in which GBP is localized at the C terminal region, and other three peptides including a putative signal peptide and appropriate processing sites for endoproteolytic cleavage precede the GBP sequence. Northern blot analyses demonstrate the presence of a 800-base mRNA transcript in fat body and 2.5-kilobase transcript in brain and nerve cord, suggesting the possibility that the transcription of GBP gene is regulated in a tissue-dependent manner. This interpretation was supported by isolating a GBP cDNA fragment from cDNA pool of brain-nerve cords. GBP mRNA is constantly expressed in both parasitized and non-parasitized last instar larvae and there is no difference in the levels of the mRNA between both larvae, thus indicating that parasitism may effect on translational or posttranslational level to elevate plasma GBP concentration. PMID- 7498539 TI - Conformational dynamics monitored by His-179 and His-200 of isolated thermophilic F1-ATPase beta subunit which reside at the entrance of the 'conical tunnel' in holoenzyme. AB - When monitored by 1H NMR at various pH values, most of the C-2 proton signals from 12 His residues of the isolated beta subunit of thermophilic F1-ATPase (TF1) could be separately observed. Two of them were assigned to His-179 and His-200 which reside at the entrance of a 'conical tunnel' to reach catalytic site in the crystal structure of F1-ATPase. His-200 gave doublet, suggesting that this region is not a rigid alpha-helix in the isolated beta subunit. The binding of Mg.AMP PNP changed the chemical shifts of His-179 and His-200 significantly. Although His-119 located at the opposite side of the conical tunnel was not affected by the nucleotide-binding, it contributed to the stability of beta subunit and the efficiency of the catalysis of the holoenzyme. PMID- 7498540 TI - Kinetics of translation of gamma B crystallin and its circularly permutated variant in an in vitro cell-free system: possible relations to codon distribution and protein folding. AB - Analysis of nascent gamma B-crystallin peptides accumulating during in vitro translation in a rabbit reticulocyte lysate cell-free system was carried out. As a consequence of the irregular distribution of rare codons along the polypeptide chain of gamma B-crystallin, translation of the two-domain protein is a non uniform process characterized by specific pauses. One of the major delays occurs during the translation of the connecting peptide between the domains. Comparing the kinetics of translation of natural gamma B-crystallin and its circularly permutated variant (with the order of the N- and C-terminal domains exchanged) reveals that the natural N-terminal domain is translated faster than the C terminal one. Since the N-terminal domain in natural gamma B-crystallin is known to be more stable and to fold faster than the C-terminal one [E.-M. Mayr et al. (1994) J. Mol. Biol. 235, 84-88], the present data suggest that the translation rates are optimized to tune the synthesis and folding of the nascent polypeptide chain. In this connection, the pause in the linker region between the domains provides a delay allowing the correct folding of the N-terminal domain and its subsequent assistance in the stabilization of the C-terminal one. PMID- 7498541 TI - The osmo-inducible gpd1+ gene is a target of the signaling pathway involving Wis1 MAP-kinase kinase in fission yeast. AB - The gpd1+ gene of Schizosaccharomyces pombe encodes an isozyme of NADH-dependent glycerol-3-phosphate dehydrogenases that is involved in glycerol synthesis, whose expression is induced upon an upshift of the medium osmolarity. We provide evidence that this osmotic induction of gpd1+ in S. pombe is under the control of a MAP-signaling pathway involving the wis1+ gene-product, which is a homologue of MAP-kinase kinases. The results suggested that the gpd1+ gene is a downstream target of the osmosensing signaling that is transmitted through Wis1, thereby defects of either of these genes result in the similar phenotype, namely, osmosensitive for growth, because of the failure in accumulation of the intracellular osmoprotectant, glycerol. PMID- 7498542 TI - Laccase-mediated formation of the phenoxazinone derivative, cinnabarinic acid. AB - The phenoxazinone chromophore occurs in a variety of biological systems, including numerous pigments and certain antibiotics. It also appears to form as part of a mechanism to protect mammalian tissue from oxidative damage. During cultivation of the basidiomycete, Pycnoporus cinnabarinus, a red pigment was observed to accumulate in the culture medium. It was identified as the phenoxazinone derivative, cinnabarinic acid (CA). Laccase was the predominant extracellular phenoloxidase activity in P. cinnabarinus cultures. In vitro studies showed that CA was formed after oxidation of the precursor, 3 hydroxyanthranilic acid (3-HAA), by laccases. Moreover, oxidative coupling of 3 HAA to form CA was also demonstrated for the mammalian counterpart of laccase, the blue copper oxidase, ceruloplasmin. PMID- 7498543 TI - Formation of 8-nitroguanine in DNA treated with peroxynitrite in vitro and its rapid removal from DNA by depurination. AB - Peroxynitrite is a strong oxidant formed by reaction of nitric oxide with superoxide in inflamed tissues. We have demonstrated that 8-nitroguanine is formed dose-dependently in calf thymus DNA incubated with low concentrations of peroxynitrite in vitro. 8-Nitroguanine in acid-hydrolyzed DNA was chemically reduced into 8-aminoguanine, which was analyzed using high performance liquid chromatography with electrochemical detection. Only peroxynitrite, but not nitrite, tetranitromethane nor NO-releasing compounds, formed 8-nitroguanine. Antioxidants and desferrioxamine inhibited the reaction. 8-Nitroguanine was depurinated from DNA incubated at pH 7.4, 37 degrees C (t1/2 = approximately 4 h). Peroxynitrite did not increase 8-oxoguanine levels in DNA. PMID- 7498544 TI - Sedimentation and immunological analyses of GLUT4 and alpha 2-Na,K-ATPase subunit containing vesicles from rat skeletal muscle: evidence for segregation. AB - In skeletal muscle insulin induces the translocation of both the GLUT4 glucose transporter and the alpha 2 subunit of the Na,K-ATPase from an intracellular membrane (IM) compartment to the plasma membrane (PM). Fractionation studies of rat skeletal muscle using a discontinuous sucrose gradient have indicated that the insulin-induced loss of both proteins occurs from a fraction containing intracellular membranes (IM) of common density. This raises the possibility that both proteins may be colocalized in a single intracellular compartment or are present in separate membrane vesicles that are of similar buoyant density. In this study we report the membrane vesicles from the insulin-responsive IM fraction can in fact be separated on the basis of differences in their sedimentation velocities; immunoblot analyses of fractions collected from a sucrose velocity gradient revealed the presence of two separate peaks for GLUT4 and the alpha 2 subunit of the Na,K-ATPase. One of these peaks representing a fast sedimenting population of vesicles (with a sedimentation coefficient of 2697 +/- 57 S) reacted against antibodies to the alpha 2 subunit of the Na,K-ATPase, whereas, the second peak contained a population of much slower sedimenting vesicles (with a sedimentation coefficient of 209 +/- 4 S) were practically devoid of the alpha 2-subunit. By contrast, the slow sedimenting vesicles were enriched by approximately 32-fold in GLUT4 relative to the starting IM fraction when the fractional protein content was taken into account. Immunoprecipitation of GLUT4-containing vesicles from the insulin-sensitive IM fraction revealed that no immunoreactivity towards either the alpha 2 or the beta 1 subunits of the Na,K ATPase could be observed, signifying that the insulin-responsive subunits of the Na,K-ATPase and GLUT4 were present in different membrane vesicles and that it was unlikely, therefore, that the insulin-induced redistribution of these proteins to the PM occurs from a common intracellular pool. PMID- 7498545 TI - Binding of the growth factor glycyl-L-histidyl-L-lysine by heparin. AB - Evidence is presented that the growth factor glycyl-histidyl-lysine (GHK) binds to heparin, and the interaction has been characterized by [1H]NMR spectroscopy. 1H chemical shifts indicate that GHK interacts with both the carboxylic acid and the carboxylate forms of heparin. The chemical shift data are consistent with a weak delocalized binding of the triprotonated (ImH+, GlyNH3+, LysNH3+) form of GHK by the carboxylic acid form of heparin. As the pD is increased and the carboxylic acid groups are titrated, chemical shift data indicate that ammonium groups of GHK are hydrogen bonded to heparin carboxylate groups, while the histidyl imidazolium ring occupies the imidazolium-binding site of heparin. Evidence for site-specific binding includes displacement of chemical shift titration curves for heparin to lower pD, increased shielding of specific heparin protons by the imidazolium ring current and displacement of chemical shift titration curves for GHK to higher pD. Specific binding constants were determined for binding of the (ImH+, GlyNH3+), LysNH3+) forms of GHK by the carboxylate form of heparin from chemical shift vs. pD titration data. PMID- 7498546 TI - Poly(U) binding activity of hepatitis C virus NS3 protein, a putative RNA helicase. AB - A non-structural protein of the hepatitis C virus (HCV), NS3, contains amino acid sequence motifs characteristic of serine-proteinases and RNA helicases. RNA binding activity of the NS3 protein with an apparent dissociation constant of 2 x 10(-7) M was detected using a poly(U)-Sepharose resin. Competitive RNA binding analysis suggested that the NS3 protein binds preferentially to the poly(U) sequence, which is located at the 3' end of HCV RNA. Mutational analysis of NS3 protein revealed the possibility that both the RNA helicase region and the serine proteinase region were necessary for full RNA binding activity. PMID- 7498547 TI - cDNA sequences of three sheep myeloid cathelicidins. AB - Several myeloid antimicrobial peptide precursors have been shown to consist of a N-terminal proregion similar to a protein named cathelin and a structurally varied C-terminal antimicrobial domain. Proteins with these features have been named cathelicidins. In this paper we report the cDNA sequences of three ovine cathelicidins of 155, 160 and 190 residues, respectively, with cationic C terminal sequences corresponding to putative antimicrobial domains. These are structurally varied and include a Cys-rich sequence of 12 residues, which is similar to the bovine antimicrobial cyclic dodecapeptide, a novel 29 residue sequence named SMAP-29 with a possible alpha-helical conformation, and a 60 residue sequence named Bac7.5, which appears to be a new member of the Pro- and Arg-rich group of mammalian antimicrobial peptides. PMID- 7498548 TI - The N-terminal, dehydrogenase/cyclohydrolase domain of yeast cytoplasmic trifunctional C1-tetrahydrofolate synthase requires the C-terminal, synthetase domain for the catalytic activity in vitro. AB - The yeast ADE3(1-333) gene which encodes a truncated protein containing the N terminal 5,10-methylene-tetrahydrofolate (THF) dehydrogenase (D)/5,10-methyl-THF cyclohydrolase (C) domain of cytoplasmic trifunctional C1-THF synthase is able to complement all the phenotypes associated with ade3 mutations in vivo. However, expression of the ADE3(1-333) gene in an ade3 strain does not retain any D activity in vitro. Expression in a yeast ade3 strain of the ADE3(1-333) fused to the Escherichia coli lacZ gene or to the yeast SER2 gene allows detection of D and C activities in vitro. These results indicate that the N-terminal D/C domain of C1-THF synthase requires the C-terminal 10-formyl-THF synthetase domain for stable catalytic activity in vitro. PMID- 7498549 TI - cDNA cloning of a human 100 kDa de-ubiquitinating enzyme: the 100 kDa human de ubiquitinase belongs to the ubiquitin C-terminal hydrolase family 2 (UCH2). AB - The full length cDNA encoding a 100 kDa human de-ubiquitinating enzyme, referred to as de-ubiquitinase was obtained using one clone selected from a randomly sequenced human brain cDNA library and specific primers. The sequence of 18 peptides generated from the de-ubiquitinase isolated from out-dated human erythrocytes matched perfectly with the predicted amino acid sequence, which would encode a protein containing 858 amino acids (calculated M(r) = 95,743 Da). Homology search disclosed that the protein is a member of a large family of ubiquitin C-terminal hydrolases (UCH2), that was defined on the basis of the presence of two specific patterns, 'the Cys- and His-domains', which are likely to be involved in the de-ubiquitinating activity [7]. An additional conserved region, 'the aspartic acid domain', was also identified, the functional role of which is unknown. PMID- 7498550 TI - In situ dephosphorylation of tau by protein phosphatase 2A and 2B in fetal rat primary cultured neurons. AB - Using antibodies recognizing the phosphorylation state of specific sites, phosphorylation states of tau were monitored in fetal rat primary cultured neurons. When cultured neurons were treated with okadaic acid (OA) or calyculin A (CalA) at concentrations sufficient to inhibit protein phosphatase 2A (PP2A), phosphorylation of Ser-199/Ser-202 (numbered according to the human tau 441) and Ser-235 increased. On the other hand, treatment with Ca2+ ionophore, A23187, induced dephosphorylation of Ser-199/Ser-202, Thr-205, Ser-396 and Ser-404, and this dephosphorylation was repressed by inhibitors of protein phosphatase 2B (PP2B), cyclosporin A and FK506. These results indicate that PP2A and PP2B are differentially involved in dephosphorylation of tau in neurons. PMID- 7498551 TI - Characterization of common neoantigenic epitopes generated in plasminogen activator inhibitor-1 after cleavage of the reactive center loop or after complex formation with various serine proteinases. AB - Plasminogen activator inhibitor-1 (PAI-1), an important risk factor for thrombotic diseases, is a member of the superfamily of serine proteinase inhibitors. To define structural rearrangements occurring during interaction between PAI-1 and its target proteinases we have raised monoclonal antibodies against the PAI-1/t-PA complex. Thirteen out of 401 monoclonal antibodies reacted preferentially with the PAI-1/t-PA complex as compared to free PAI-1 or free t PA. Detailed characterization revealed the presence of two non-overlapping neoantigenic epitopes in the PAI-1/t-PA complex. Both neoantigenic epitopes were also exposed after complex formation between PAI-1 and either urokinase-type plasminogen activator, plasmin or thrombin as well as after cleavage of the reactive site loop of non-inhibitory substrate type PAI-1 variants. Thus, we have identified two neoantigenic epitopes, localized entirely in PAI-1, and commonly exposed after complex formation of active PAI-1 with various proteinases or after cleavage of substrate PAI-1. These monoclonal antibodies should facilitate further studies on the mechanism of interaction between various PAI-1 forms and its target proteinases. PMID- 7498552 TI - Activin A-induced apoptosis is suppressed by BCL-2. AB - Activin A, a member of TGF beta superfamily has various activities including induction of apoptosis in mammalian cells. However, it remains unknown how activin A induces cell death. To clarify this, we investigated the expression of BCL-2 and BAX, and the effect of BCL-2 overexpression on activin A-induced apoptosis in B cell hybridoma cell lines. The activin A-sensitive cell lines expressed BAX but not BCL-2 and that activin A did not increase BAX levels. Overexpression of human BCL-2 suppressed activin A-induced apoptosis in these cells. Thus, activin A has been shown to induce apoptosis by a BCL-2-inhibitable mechanism without activating BAX. PMID- 7498553 TI - Insect prothoracicotropic hormone: a new member of the vertebrate growth factor superfamily. AB - Prothoracicotropic hormone (PTTH) is a brain neurosecretory protein that controls insect development. PTTH of the silkmoth Bombyx mori is a homodimeric protein, the subunit of which consists of 109 amino acids. Clear-cut sequence similarity to any other proteins has not been observed. By disulfide-bond pattern analysis and modeling of the PTTH structure based on the known three-dimensional (3D) structures of growth factor family with cystine-knot motif, we propose that the PTTH protomer adopts the fold unique to the structural superfamily of the growth factors, beta-nerve growth factor (beta-NGF), transforming growth factor-beta 2 (TGF-beta 2), and platelet-derived growth factor-BB (PDGF-BB). The insect neurohormone PTTH appears to be a member of the growth factor superfamily, sharing a common ancestral gene with the three vertebrate growth factors, beta NGF, TGF-beta 2 and PDGF-BB. PMID- 7498554 TI - Cloning of a Syrian hamster cDNA related to sexual dimorphism: establishment of a new family of proteins. AB - The clone FHG22, isolated from a female minus male subtracted cDNA library obtained from the sexually dimorphic Syrian hamster Harderian glands (HG) is 440 bp long with a 95 amino acids ORF, and hybridizes to a female HG-specific 0.6 kb mRNA. The FHG22 nucleotide and amino acid sequences are similar to the subunits from prostatein, uteroglobin, major cat allergen Fel dI (chain 1) and mouse salivary androgen binding proteins (subunit alpha). Therefore I propose that all those polypeptides belong to a common new family. The hamster genome has a single copy of the FHG22 gene, without homologous genes. FHG22 mRNA is also found in male and female parotid (higher levels in females) and submandibular glands, indicating a tissue and sex-dependent control of expression. PMID- 7498555 TI - Novel guanylyl cyclase inhibitor, ODQ reveals role of nitric oxide, but not of cyclic GMP in endothelin-1 secretion. AB - The role of nitric oxide (NO) and guanosine 3',5'-cyclic monophosphate (cyclic GMP) in cellular regulation of endothelin-1 (ET-1) secretion was investigated in cultured porcine aortic endothelial cells. NO synthase was inhibited with NG nitro-L-arginine (L-NNA) and guanylyl cyclase with the novel selective inhibitor, ODQ (1H-[1,2,4]oxadiazolo[4,3-a]quinoxalin-1-one) (3 microM). Basal and phorbol ester (PMA)-stimulated ET-1 secretion were unaffected by ODQ, but stimulated secretion was increased by L-NNA. In the presence of the NO donors, spermine/NO, S-nitroso-glutathione (GSNO), and nitroprusside (NP) ET-1 secretion was reduced, but ODQ had no effect on this inhibition, although it effectively inhibited cyclic GMP production. NO release from donors, measured with a sensitive NO electrode, was greatest for spermine/NO, intermediate for GSNO, minimal for NP and paralleled inhibition of ET-1 secretion. The data suggest that in cultured endothelial cells, curtailment of ET-1 secretion is mediated by NO and independent of cyclic GMP. PMID- 7498556 TI - Let there be light? PMID- 7498557 TI - Topical chloramphenicol: use or abuse? PMID- 7498558 TI - The Bowman Lecture. PMID- 7498561 TI - Intraocular deposition of metallic fragments during phacoemulsification: possible causes and effects. AB - We undertook a prospective slit lamp examination of 56 eyes which had undergone routine phacoemulsification, in order to determine the incidence of metallic intraocular foreign bodies. Characteristic small reflective particles were noted in 86%, with a mean of 5.1 particles per affected eye. No accompanying clinical problems were seen. The used phacoemulsification needles were examined with colour photography and scanning electron microscopy. A characteristic pattern of wear was observed, corresponding to an area under the plastic irrigating sleeve rather than at the tip. This suggests cavitation erosion rather than instrument contacts as the cause. Titanium alloy contains 6% aluminium, 4% vanadium and 0.25% iron. All its constituents may corrode and leach from the surface of the metal when in contact with body fluids and have been implicated in numerous adverse biological processes. PMID- 7498559 TI - Systemic cyclosporine in high-risk keratoplasty: long-term results. AB - Systemic cyclosporine (CSA) was given to 43 high-risk keratoplasty patients (vascularisation in three or four quadrants and > 16 stromal vessels): 14 received CSA for 12 months (long CSA group) and 29 for a 4-6 month period (short CSA group). A group of 37 similar high-risk keratoplasty patients received no CSA (no CSA group). In the no CSA group 27 (73%) grafts had rejection episodes compared with 21 (48.8%) in the combined CSA group (p = 0.025). Rejection was reversed in only 23.3% of the no CSA group, compared with 50% and 87.5% in the short (p = 0.06) and long (p = 0.002) CSA groups respectively. Compared with the no CSA group, overall graft survival was better in both the short (p = 0.019) and long (p = 0.0056) CSA groups. This improvement resulted from both a reduction in the incidence, and an increase in the reversal rate, of rejection episodes. The improvement continued after stopping CSA, suggesting that some immunological privilege had been re-established. Acuities of 20/40 or better were achieved by 44.8% and 50% of eyes in the short and long CSA groups respectively, compared with 13.5% of eyes in the no CSA group. PMID- 7498560 TI - Over-expression of a gelatinase A activity in keratoconus. AB - Keratoconus is an ocular disorder in which the central cornea becomes thin, conical and frequently scarred. We are exploring the possibility that this condition is induced and maintained by proteases that exist in the corneal matrix in an activated form. In this study, the activities of the proteases secreted in vitro and in vivo by keratocytes of normal, clear keratoconic, scarred keratoconic and traumatically scarred corneas have been compared and partially characterised. Data obtained by assaying acyl transferase activity showed that the matrix metalloproteinases account for a minimum of 95% of the total protease secreted by cultured keratocytes. Their summated specific activity was consistently and significantly higher in the culture medium of keratoconic keratocytes than in the medium of other keratocyte cultures. Analysis of the individual protease activities secreted by these corneal keratocytes in vitro and in vivo by SDS-gelatin polyacrylamide gel electrophoresis showed that a gelatinase of molecular weight 65,000 is the major protease secreted by normal keratocytes. Whereas clear keratoconic and traumatically scarred corneal keratocytes secrete an additional activity of molecular weight 61,000, scarred keratoconic corneal keratocytes generally produced little or none of this gelatinase activity. Both activities may be ascribed to gelatinase A, and although the 61,000 molecular weight form may be a significant feature of keratoconus, neither appears to be active as secreted. PMID- 7498562 TI - Dislocated nuclear fragments after cataract surgery. AB - Posterior nuclear dislocation is a serious complication of cataract surgery, especially when using the phacoemulsification technique. So far, there have been only a few reports concerning the indications and timing of vitrectomy with nuclear removal as well as long-term visual outcome of these eyes. We analysed 23 consecutive patients (follow-up at least 3 months) with intravitreal nuclear remnants after cataract surgery treated with vitrectomy and removal of posteriorly dislocated nuclei. All the eyes had increased intraocular pressure pre-vitrectomy, 63% had corneal oedema, 67% marked uveitis and 26% either retinal tear or detachment. Vitrectomy was performed within 1 week in 70% of eyes. In 14 eyes (61%) the final visual acuity was 20/40 or more. The main reason for poor visual outcome was retinal detachment (9%). These results indicate that with vitrectomy and removal of the nucleus good visual results can be achieved in a large proportion of eyes with posterior dislocation of nuclear remnants. PMID- 7498563 TI - Implantation of foldable intraocular lenses in the presence of anterior capsular tears. AB - A consecutive series of patients who had undergone small-incision phacoemulsification cataract surgery with foldable lens implantation were reviewed. Seven patients had sustained anterior capsular tears during surgery. In all cases a three-piece foldable intraocular lens (IOL) was successfully inserted by careful positioning of the haptics. In all 6 patients available for long-term assessment the IOL remained stable and well centred, the tears having been sealed by fibrous tissue. We show that, if logical measures are taken to limit tear extension, the lack of an intact capsulorhexis need not result in conversion to extracapsular cataract extraction. PMID- 7498564 TI - Vitreous loss in planned extracapsular cataract extraction does lead to a poorer visual outcome. AB - Forty-six cases of vitreous loss during planned extracapsular cataract extraction salvaged by anterior vitrectomy and primary anterior chamber lens implantation were compared in a retrospective cohort study with 92 individually matched controls. The postoperative visual outcome for cases with vitreous loss was significantly poorer than that for controls even after adjustment for confounding variables. Part of this visual morbidity was explained by cystoid macular oedema and retinal detachment, but even after adjusting for these two complications visual outcome remained worse among cases than controls. There was a significantly higher incidence of clinically apparent cystoid macular oedema amongst cases compared with controls (20% versus 1%). Two retinal detachments occurred among the cases but this difference in incidence was not statistically significant. PMID- 7498565 TI - Phenylephrine and pilocarpine in the treatment of post-operative irido-corneal adhesion. AB - Following cataract surgery, entrapment of the iris within the surgical wound is often managed by intensive use of miotics. As the radial fibres stretch, only a small amount of fraction is exerted upon the entrapped iris. Application of a combination of phenylephrine and pilocarpine drops causes simultaneous contraction of the pupil sphincter and the radial muscle fibres. This study investigated the relative magnitude of forces induced in the iris periphery by pilocarpine and phenylephrine and the effectiveness of adding g. phenylephrine 10% to g. pilocarpine 4% drops in the treatment of postoperative irido-corneal adhesions. The investigation was divided into two parts. First, the forces induced in the iris periphery upon exposure to pilocarpine and phenylephrine were measured in 6 cadaver irises. The mean force was 27.5 +/- 5.7 x 10(-3) N for pilocarpine and 23.3 +/- 4.0 x 10(-3) N for phenylephrine. The combination of the two drugs produced a force of 54.2 +/- 6.6 x 10(-3) N (p < 0.05). In the second part of the study intensive pilocarpine 4% drops were administered to 17 patients who had iris-wound entrapment on the first post-operative day. Patients with persistent adhesion were commenced on intensive g. phenylephrine 10% and assessed after 90 minutes. Of the 17 patients, 6 responded to pilocarpine drops alone; in a further 7 the irido-corneal adhesion was released only by the addition of phenylephrine drops, and in 4 patients drops were ineffective in relieving the adhesion. This study indicates that addition of phenylephrine 10% to pilocarpine 4% drops enhances the effectiveness of pharmacological treatment of post operative irido-corneal adhesion. PMID- 7498566 TI - Topical anaesthesia for phacoemulsification surgery. AB - Although phacoemulsification with only topical anaesthesia is possible, the level of any discomfort perceived by the patient is unknown. Topical anaesthesia eliminates any risk of inadvertent ocular or orbital injury. Twenty-four patients undergoing phacoemulsification under only topical anaesthesia (g. amethocaine 1%) were compared with 23 patients undergoing the same procedure with peribulbar anaesthesia (5 ml lignocaine 2%, 5 ml bupivacaine 0.5% and 75 units hyaluronidase). Perceived pain on administration of the anaesthesia, per operatively and post-operatively was assessed by a nursing officer using a visual analogue scale (0-10). It was found that there was no difference in pain score between the two groups on induction of anaesthesia. Per-operatively and post operatively scores were higher in the topical group than in the peribulbar group (p < 0.05, Mann-Whitney U-test), though most of these were within the range 0-3, ('no pain' to 'slight pain'). There was no difference in the amount of analgesia dispensed to the two groups on the ward post-operatively. In conclusion, topical anaesthesia provides adequate anaesthesia for phacoemulsification. Although a small increase in the level of discomfort was observed, the pain levels reported were small and may not be clinically significant when set against the reduced incidence of anaesthetic-related complications. PMID- 7498567 TI - Success rate and complications of intraoperative 0.2 mg/ml mitomycin C in trabeculectomy surgery. AB - Adjunctive chemotherapy with Mitomycin C (MMC) has been used in an attempt to modulate the wound healing response in glaucoma filtration surgery. A consecutive series of 20 eyes from 18 patients undergoing trabeculectomy with MMC intraoperatively was studied. Sixteen cases were considered high risk regarding surgical success and 4 patients with low tension glaucoma (LTG) required lower intraocular pressure (IOP) to prevent further visual field loss. Surgical technique involved the use of a limbal-based conjunctival flap and MMC 0.2 mg/ml applied via a sponge (under the scleral flap) to both scleral and conjunctival surfaces for 5 minutes. The mean follow-up period was 12.7 months (range 3-24). There were 17 successful eyes. Of these, 14 are high pressure glaucoma eyes with a mean pre-operative IOP of 30.9 +/- 10.9 mmHg and a mean postoperative IOP of 15.3 +/- 5.2 mmHg (p = 0.001). The remainder of the successful cases include 4 patients with LTG with a mean preoperative IOP of 17.8 +/- 0.5 mmHg and a mean postoperative IOP of 6.8 +/- 0.7 mmHg (p = 0.001). Serious complications included chronic repeated bleb leaks (n = 2) and scleral necrosis (n = 2). There was one case of hypotonous maculopathy. These results are comparable with those of other studies. Despite a relatively low dose of MMC serious side-effects were encountered. Management of these complications is described, and how these effects may be prevented by altering scleral exposure to MMC. In addition a possible explanation for the serious side-effects of MMC-treated trabeculectomies is presented. PMID- 7498569 TI - Greater severity of clinical features in older patients with Duane's retraction syndrome. AB - Duane's retraction syndrome (DRS) is a well-described entity. However, the possibility of this condition having progressive features has not been previously documented. We studied 21 consecutive patients with this disorder and found that the incidence of severe retraction on adduction, the occurrence of enophthalmos in the primary position and the presence of upshoots and downshoots were significantly higher in adults with type I DRS than in children. Awareness that the clinical features may increase in severity is important as parental counselling, coupled with effective communication to general practitioners, can prompt re-referral should such problems arise. Surgical intervention as a consequence, becomes an available option to correct disease progression. PMID- 7498568 TI - 'Henderson's' relief of eyelid retraction revisited. AB - The purpose of the study was to identify those patients with thyroid eye disease and upper eyelid retraction who would benefit from Henderson's procedure (myotomy of Muller's superior tarsal muscle with graded division of levator aponeurotic fibres on the anterior tarsal plate). The clinical records of patients attending the Thyroid Clinic of Moorfields Hospital who had undergone Henderson's procedures were reviewed. Twenty-two patients (19 women, 3 men) had Henderson's procedures at age 35-69 years (mean 49.9), with a mean follow-up of 4.55 years. Eleven patients had bilateral upper eyelid surgery, 11 unilateral (33 eyelids). The pre-operative vertical palpebral apertures ranged from 11 to 19 mm (mean 13.95); the post-operative vertical palpebral apertures ranged from 8 to 15 mm (mean 11.45), a mean reduction of 2.5 mm (p < 0.001). Eight of twenty-two patients (36%) had further eyelid surgery and 3 of 22 (14%) had further surgery recommended because of persistent upper eyelid retraction, abnormal eyelid contour (lateral flare) and asymmetry. It is concluded that Henderson's procedure should be reserved for those patients with minor degrees of almost symmetrical upper eyelid retraction who do not have abnormalities of their eyelid contour. PMID- 7498570 TI - Spontaneous recovery rates for unilateral sixth nerve palsies. AB - Two hundred and thirteen patients with unilateral isolated sixth nerve palsies were assessed to determine what proportion of them underwent spontaneous recovery and over what period of time this recovery occurred. All were primary ophthalmic referrals from which trauma was excluded. In all, 78.4% of patients experienced spontaneous recovery of their palsy, 36.6% recovering by 8 weeks and 73.7% by 24 weeks. Only 16.4% failed to recover. Of this group, however, nearly 40% had serious underlying pathology accounting for their palsy. PMID- 7498571 TI - Midline nasal dermoids presenting like discharging lacrimal sac mucocoeles. AB - Two cases of midline intranasal dermoid cysts presenting like discharging lacrimal sac mucocoeles in children are described. The diagnosis of this rare condition is suggested by the presence of midline nasal cutaneous dimples, although these may be associated with isolated anomalies such as nasal dermoid cyst, encephalocoele or glioma. PMID- 7498573 TI - Ultrasonographic investigation of optic neuritis. AB - A pilot echographic study was conducted to determine the incidence and severity of optic nerve swelling in acute 'idiopathic' optic neuritis and to examine cerebrospinal fluid dynamics in the subarachnoid space, employing the method of standardised echography and the '30 degrees test'. An attempt was made to correlate the degree of nerve swelling with the initial visual loss and with the rate and extent of recovery of vision. The visual function and echographic features of the optic nerve in 27 patients with the diagnosis of acute optic neuritis were assessed with standardised echography. A significant increase in nerve diameter was found in 74% of cases. There was a correlation between nerve swelling and the severity of initial visual loss. The authors conclude that standardised echography is a useful tool in the diagnosis of optic neuritis and may play a role in predicting the visual outcome. PMID- 7498572 TI - Endonasal laser dacryocystorhinostomy under direct vision. AB - We describe a method of endonasal laser dacryocystorhinostomy, using a neodynium YAG laser, whose ease of performance should make it attractive to general ophthalmologists as an alternative to the routine external approach. A small series of 6 patients were investigated 6 months after surgery and a patent drainage system was revealed. PMID- 7498574 TI - Doppler ultrasonography of the central retinal artery in patients with diabetes and vascular disease treated with topical timolol. AB - Using high-resolution duplex scanning it is possible to evaluate the blood flow velocity in the central retinal artery of the eye. Four different patient groups were studied with this technique: normals, diabetics with a decreased flow, diabetics with an increased flow and vascular patients with a decreased flow. The eyes of these patients were then treated with topical timolol and the flow measured again. An increase in the flow was found in three of the four groups. This increased flow velocity may be due to a vasodilatory effect of timolol. The results are presented and discussed. PMID- 7498575 TI - The use of dynamic ultrasound B-scan to detect retinal tears in spontaneous vitreous haemorrhage. AB - The use of dynamic ultrasound B-scan to detect retinal tears in spontaneous vitreous haemorrhage is described. A 6-month prospective study was undertaken of non-diabetic patients who presented with spontaneous vitreous haemorrhage, in whom the fundus could not be visualised and in whom a rhegmatogenous aetiology was suspected. Patients were followed up at least weekly with repeat B-scans until adequate fundus visualisation was possible. Eight patients in total were included in the study; of these, 4 were thought to have retinal tear at initial ultrasound examination and this was confirmed later when the haemorrhage cleared. A scanning protocol is suggested. PMID- 7498576 TI - Thrombophilic factors in ischaemic and non-ischaemic idiopathic retinal vasculitis. AB - Two common causes of visual loss in idiopathic retinal vasculitis (RV) are retinal ischaemia and cystoid macular oedema. This study investigated whether thrombophilic factors are more prevalent in patients with ischaemic RV than non ischaemic RV. Twenty patients with RV (10 ischaemic, 10 non-ischaemic) were prospectively recruited before starting systemic immunosuppression. Twenty-one different haemostatic parameters were tested. Seventeen patients had at least one haemostatic abnormality. Three patients had low Protein S, one had low Protein C. Three patients had positive anticardiolipin antibody titres, 1 had poor fibrinolytic activity, 3 had raised fibrinogen levels. Ten patients had raised lipoprotein (a) levels. Fibrinogen levels were higher in the smokers (p = 0.02). Although all von Willebrand's factor levels were within the normal range, they were higher in the ischaemic group (p = 0.008), in which smoking was more prevalent. This study has shown a high prevalence of thrombophilic abnormalities in RV patients, and implicates smoking in the aetiology of ischaemic RV. PMID- 7498577 TI - Leber's hereditary optic neuropathy: implications of the sex ratio for linkage studies in families with the 3460 ND1 mutation. AB - Leber's hereditary optic neuropathy (LHON), which is associated with mutations in mitochondrial DNA (mtDNA), is commoner in males than females. A study of over 30 LHON families with a mutation at position 3460 of mtDNA demonstrates a significantly decreased male excess from that generally quoted, with evidence for a marked bias in the ascertainment of males over females. This has implications for the analysis of those factors which give rise to the male bias. PMID- 7498579 TI - How to master capsulorhexis. PMID- 7498578 TI - How much blame can be placed on laser photocoagulation for failure to attain driving standards? AB - One hundred consecutive patients who underwent bilateral pan-retinal photocoagulation (PRP) for proliferative diabetic retinopathy were assessed in accordance with the UK Driver and Vehicle Licensing Agency (DVLA) guidelines. Visual acuity was documented, and visual fields were assessed using the Esterman test. Among the 30% of patients who failed to reach the visual standards required for a driving licence, three groups were identified: those who failed to attain either the required binocular visual acuity (n = 4), or visual fields (n = 9), or both (n = 17). Previous studies reveal a large variation in DVLA field test failure following PRP treatment for proliferative diabetic retinopathy. Our results show a 19% failure rate solely attributable to treatment, which is at the lower end of previously reported studies (20-80%). The reasons for this discrepancy are discussed. We conclude that modern treatment procedures for proliferative diabetic retinopathy may be undertaken with the knowledge that in the majority of cases a patient's driving licence is unlikely to be revoked. PMID- 7498580 TI - Persistent tachycardia following subconjunctival injections of mydriatic agents (Mydricaine) used for maintenance of perioperative mydriasis in vitreoretinal surgery. PMID- 7498581 TI - Lattice dystrophy and corneal ulceration. PMID- 7498582 TI - Administration of eye drops in the community: the best use of a district nurse's time? PMID- 7498583 TI - The management of an optic nerve compromising acute retrobulbar haemorrhage: report of a case. PMID- 7498584 TI - Endophthalmitis caused by Enterococcus faecalis. PMID- 7498586 TI - Adherence of silicone oil to intraocular lenses. PMID- 7498585 TI - New forceps for implanting intraocular lenses. PMID- 7498588 TI - Uveitis and skin tattoos. PMID- 7498587 TI - Vitreous loss during cataract surgery. PMID- 7498589 TI - Dynamic mechanical analysis of two crosslinked copolymer systems. AB - OBJECTIVES: This research investigated the dynamic mechanical properties of two model dental restorative copolymers to elucidate the influence of pendent group length and addition of crosslinking agent. METHODS: Two different monomethacrylate resins [(methyl methacrylate (MMA) or 2-ethoxyethylmethacrylate (EEM)] were added to various molar fractions (0, 5, 10, 20, 40, 60, 80 and 100%) of a difunctional monomer [triethylene glycol dimethacrylate (TEGDMA)], resulting in two different copolymer systems (TMMA and TEEM). Bar-shaped specimens were heat-cured and tested for their dynamic mechanical properties at a fixed frequency (1Hz): flexural storage modulus and loss factor. Transition temperatures associated with peaks were determined using tan delta, phase angle and loss factor. The properties of one specimen were also examined using a range of applied frequencies. RESULTS: Below the Tg, all resin systems had similar storage moduli. An increase in pendent group length lowered transition temperatures and demonstrated greater influence on temperature changes than did a change in crosslinker content. With increasing crosslinker, the transition temperatures increased. The values ascribed to transition temperatures were dependent upon the analysis method used: tan delta gave highest values, followed by phase angle, with loss factor values the lowest. Storage and loss moduli values and their ratio increased with increasing frequency of applied signal. Loss factor analysis provided the most elucidation of sub-Tg transitions. SIGNIFICANCE: Restorative polymers with specific desired physical and mechanical properties can be achieved by both judicious selection of monomer structure as well as by the controlled addition of the crosslinking agent. PMID- 7498590 TI - Correlation of cytotoxicity with element release from mercury- and gallium-based dental alloys in vitro. AB - OBJECTIVES: An in vitro screening test was used to compare the cytotoxicity and elemental release from mercury- and gallium-based dental restorative materials. METHODS: The test employed three sequential extractions of the samples into cell culture medium which were then used to evaluate the cytotoxicity of the samples and the release of elements from the samples. Cytotoxicity was measured by placing the extract in contact with Balb/c mouse fibroblasts for 24 h and measuring the succinic dehydrogenase activity of the cells. The release of elements was measured by means of atomic absorption spectrophotometry. RESULTS: Samples of Tytin (Kerr) showed no cytotoxicity compared to Teflon controls. Dispersalloy (Johnson and Johnson) was severely cytotoxic initially when Zn release was greatest, but was less toxic between 48 and 72 h as Zn release decreased. Gallium Alloy GF (Tokuriki Honten) was moderately cytotoxic after 8 h, and increased in cytotoxicity thereafter, which correlated with a substantial and persistent release of Ga from this material. SIGNIFICANCE: The results of the current study concurred with in vivo assessments of these materials, and the use of sequential extractions was useful in determining trends in the cytotoxicity and elemental release from these materials. PMID- 7498591 TI - Biological and clinical investigations of a glass ionomer base material. AB - OBJECTIVES: The purpose of this study was to investigate the biological properties of a new glass ionomer base material containing strontium glass. METHODS: The glass ionomer base material (Base Line, De Trey Dentsply) was tested for cytotoxicity using a standard agar diffusion test, for its effect upon the pulp in subhuman primates, and in a clinical study as base beneath both composite fillings and amalgam fillings. RESULTS: The material showed a cytotoxicity pattern similar to reference materials, a zinc phosphate cement and a reference glass ionomer base material, which are used successfully in clinical applications. With the pulp/dentin usage test in subhuman primates, there were no statistically significant differences between the experimental glass ionomer base material and the negative control group if bacterial contamination of the cavity floor or direct contact with the pulp was avoided. The results of the clinical study also indicated no pulp damage after one year. SIGNIFICANCE: The experimental glass ionomer base material proved to be acceptable both biologically and clinically. PMID- 7498592 TI - Sensitivity of catalyst/base ratio on curing of resin luting agents: polymerization exotherm analysis. AB - OBJECTIVES: Currently, the proposed test of the International Standardization Organization (ISO) for measuring working and setting times of resin luting agents is based on measurement of times to reach specified stages on the polymerization exotherm. The objective of this study was to use this test to investigate the influence of variations in the mass ratios of catalyst paste to base paste on the working and setting times of three dual-cured dental resin luting agents. METHODS: The materials used were Dicor Light Activated Cement (Dentsply International Inc.), Palfique Inlay Resin Cement (Tokuyama Soda Co.), and Vivadent Dual Cement (Vivadent). Fifteen specimens of each material were tested for working time by spatulating mass ratios from 0.7 to 1.3 for 30s at 23 degrees C and recording the time from beginning of spatulation to the time at which a temperature increase occurs. Ten specimens of each material were tested for setting time by spatulating in a similar manner at 37 degrees C and recording the time at which the temperature reaches a maximum value. RESULTS: The data were fitted to the relation, In t = In A + Bm, where t is the time in seconds, m is the mass ratio, and A and B are regression coefficients. The results suggest that working and setting times of the specimens were independent of variations in mass ratio. A comparison among the materials was made by using a multiple linear regression with the relation, In t = In C + Dm + E gamma + Fm gamma, where gamma is a dummy variable to help distinguish between materials, and C, D, E, and F are regression coefficients. The results suggest that differences in materials influence the working time but not the setting time. SIGNIFICANCE: These results infer that variations in mass ratio (+/- 20%) often observed in the clinical setting should not have a significant influence on the working and setting times of resin luting agents. PMID- 7498594 TI - Effects of finishing and polishing procedures on the surface texture of resin composites. AB - OBJECTIVES: This study was designed to investigate the effects of finishing and polishing procedures on the roughness and color of resin composite surfaces. METHODS: Four composites and three different polishing systems were evaluated. Forty disc (10 mm x 2 mm) specimens of each composite were prepared. Thirty of them were ground wet through 320-grit paper, and the ground surfaces were used as the base line for the finishing procedure. Ten Mylar-covered specimens were used as controls. A color meter was applied to determine the color differences and a roughness meter was used to measure the surface roughness (Ra). RESULTS: Microfilled composite demonstrated significantly less roughness (p < or = 0.01) than hybrid composite systems as tested by ANOVA and Scheffe's tests. Polishing procedures produced a decrease in the roughness, ranging from 26 to 74%. Relative to the shade guide, the mean color difference values for all the composites after polishing were significantly greater than for the controls (p < or = 0.01) and ranged from 1.08 to 8.15 units. A positive relationship between mean color differences and Ra values was determined by regression analysis (r = 0.55). SIGNIFICANCE: A Mylar strip can create a smoother surface than the other types of tested polishing procedures. Because of the greater values in color differences and surface roughness, the shades of tested resin composites were lighter after the polishing procedures. A custom-made shade guide is suggested based on the results of this study. PMID- 7498595 TI - New adhesives in Class V restorations under combined load and simulated dentinal fluid. AB - OBJECTIVES: This study compared the efficacy of three dentinal adhesives using the "all etch" technique (All-Bond 2, Bisco; Scotchbond MP,3M Dental Products Co.; OptiBond, Kerr) with a dentinal adhesive which still uses phosphoric acid to condition enamel and a self-etching primer for dentin (A.R.T.-Bond, Coltene/Whaledent). METHODS: Eight V-shaped mixed Class V restorations were placed per group in extracted human premolars. The restorations were subjected to 1,200,000 mechanical occlusal cycles (max. force 49 N; frequency 1.7 Hz) and 3,000 simultaneous thermal cycles (5-50-5 degrees C). Dentinal fluid was simulated using 1:3 diluted horse serum and fed into the pulp chamber both during restoration and loading. Percentages of "continuous margin" were assessed on SEM replicas of enamel and dentinal margins at 200x magnification immediately before and after stressing, respectively. RESULTS: No significant differences were observed before stress between the materials either in enamel or in dentin. After stress, however, OptiBond and A.R.T.-Bond performed significantly better in dentin than the two other adhesives (Kruskal-Wallis, Mann-Whitney; p < 0.05). Although high initial values were observed, All-Bond 2 and Scotchbond MP were not stress-resistant under simulated physiological conditions. SIGNIFICANCE: The predicted clinical potential of All-Bond 2 and Scotchbond MP is inferior to that of OptiBond and A.R.T.-Bond. PMID- 7498593 TI - Clinical evaluation of four dentin bonding agents in Class V abrasion lesions: a four-year follow-up. AB - OBJECTIVES: The purpose of this study was to clinically evaluate the long-term retention of four dentin bonding agents in Class V abrasion/erosion lesions during a four-year period. METHODS: Two hundred twenty-three restorations were placed in 81 patients. Four bonding agents and composite resins (Mirage bond/Silux, Tenure/Opalux, Tripton/Opalux and Scotchbond 2/Silux) were placed according to the manufacturers' instructions. No additional undercuts or etching of adjacent enamel was done. The restorations were evaluated every six mon during a four-year period. RESULTS: Two restorations developed secondary caries during the study. After six mon, three y, and four y, respectively, the loss rates for the dentin bonding agents were: Mirage bond, 38.6%/73.0%/74.6%; Tenure, 8.5%/51.2%/56.1%; Tripton, 30.2%/83.0%/83.0%; and Scotchbond 2, 9.6%/21.2%/23.1%. Scotchbond 2 showed significantly fewer failures than the other three bonding agents. No correlation was found between retention failure frequency and age of the patients. After two years, 88.0% of all the retention failures were registered. SIGNIFICANCE: Despite the promise of dentin bonding agents to form a resin-reinforced layer in the dentin, a high clinical failure rate was registered for three of the four evaluated bonding agents in Class V restorations. PMID- 7498596 TI - The effect of irradiation time on the shear strength of composites. AB - OBJECTIVES: The purpose of this study was to determine the shear strength of composites at a specific depth by a double shear test and to relate it to irradiation time. METHODS: Aluminum molds filled with three different composites were irradiated for the manufacturer's recommended time, as well as for three longer and two shorter times. Fifteen cylindrical specimens were prepared for each combination of material and exposure time and subjected to a double shear test based on the principle of rivets connecting various structural members. The shear planes were symmetrically located 1.5 mm from both outer surfaces. RESULTS: Shear strength vs. irradiation time was directly but nonlinearly related. All test materials attained a maximum shear strength value which was not exceeded by a further increase of the exposure time. Maximum values obtained were 42.5 MPa for Heliomolar (Vivadent) and Durafill (Kulzer) and 66.9 MPa for P-50 (3M). Only the shear strengths using the shortest light activation time were significantly different for each product. SIGNIFICANCE: Shear strength of composites at a specific depth is a function of the combination of light energy and the attenuating power of the specific material. A double shear test might serve as an adjunct to microhardness in determining the extent of cure of composites at a specific depth. The advantage of this testing is the extra information to be gathered regarding the ability of composites to withstand internal stresses at a predetermined distance. PMID- 7498597 TI - Development of high-toughness resin for dental applications. AB - OBJECTIVES: The purpose of this study was to develop a heat-cured resin with improved toughness. Polyurethane dimethacrylate (PUDMA) with hard and soft segments in the chemical structure was synthesized using polyurethane diisocyanate and 2-HEMA. METHODS: The bulk polymerization of PUDMA, Bis-GMA, TEGDMA and MMA was carried out in a glass tube with 0.5 wt% of BPO at 60 degrees C for 24 h and 110 degrees C for 3 h. The physical and mechanical properties of these polymers were measured, i.e., the polymerization shrinkage, water sorption, transverse strength, modulus of elasticity, deflection and Knoop hardness of these polymerized methacrylates were determined. RESULTS: The volume shrinkage and the water sorption values of PUDMA were lower than those of Bis-GMA. The transverse strength and the modulus of elasticity of PUDMA were close to those of PMMA. The deflection of PUDMA, as determined by a bending test, was higher than any of the other cured resins tested. SIGNIFICANCE: These results suggest that PUDMA is an excellent toughened polymer. The fracture toughness of PUDMA is sufficient for crown and bridge resins and dental composites. PMID- 7498598 TI - Flexural behavior of visible light-cured composites as a function of temperature under water immersion test conditions. AB - OBJECTIVES: The purpose of this study was to investigate the effect of temperature of the flexural behavior for four visible light-cured hybrid composite materials. METHODS: Light-cured samples were post-cured for 24 h at 37 degrees C in 100% relative humidity prior to testing. Flexural tests were performed at the following water immersion temperatures: 5, 15, 25, 37, 45 and 55 degrees C, as well as in the dry condition which served as control. The following flexural property parameters were obtained as a function of test temperature: flexural strength, sigma fs, flexural modulus, Efs, flexural stress at 0.06% total offset strain, 0.06% (yield stress), and the total displacement at fracture, delta fs. Statistically significant differences (p < 0.01) in flexural parameters were noted for all the materials as a function of test temperature. RESULTS: The yield stress, sigma 0.06% was found to be most sensitive to thermomechanical changes. The results of this study indicate significant softening due to temperature under wet immersion test conditions for all materials. The temperature range at which two of the materials show significant reduction in yield stress compare well with "Wet Tg" values obtained by dynamic thermal mechanical analysis (Culbertson et al., 1990; 1991). SIGNIFICANCE: The effect of damage accumulation on surface or subsurface microstructure due to transient thermomechanical loads needs to be taken into account in the correlation of in vitro and in vivo wear or creep behavior. PMID- 7498599 TI - Comparison of the color stability of ten new-generation composites: an in vitro study. AB - OBJECTIVES: The aim of this in vitro study was to evaluate the color stability of modern light-cured composites when subjected to various physico-chemical and staining conditions. METHODS: Ten brands were evaluated including hybrids, microfine hybrids and microfilled composites. Some universal shade samples underwent only staining tests, while others were subjected to one of the following experimental conditions: thermocycling, postcuring, polishing or a 1 wk immersion in saline, prior to staining. The coloring solutions used for the staining tests were: coffee, E 110 food dye, vinegar and erythrosin. A colorimetric evaluation according to the CIE L*a*b* system was performed after experimental periods of 1 and 3 wk. RESULTS: Erythrosin caused the greatest color change for the composites tested. A reduced susceptibility to staining was observed where surfaces had been polished. Low water sorption, a high filler resin ratio, reduced particle size and hardness, and an optimal filler-matrix coupling system were related to improved composite resistance to discoloration. SIGNIFICANCE: Resistance of modern composites to discolorations still depends on their structure and manipulation. PMID- 7498600 TI - Efficacy of various commercial dentin bonding systems. AB - OBJECTIVES: The purpose of this study was to test the bonding efficacy of fifteen commercial dentin bonding systems and also to determine the effect of various commercial dentin cleaners on bonding and dentin hardness. METHODS: Each commercial bonding system was examined by two methods: 1) measuring the maximum contraction gap of a light-cured composite in a cylindrical dentin cavity; and 2) determine the tensile bond strength to a flat dentin surface. In addition, various commercial dentin cleansers were investigated to determine if dentin hardness was reduced after cleaning. As a control, the dentin surface was cleaned with neutralized EDTA prior to the combined application of an experimental dentin primer, a commercial bonding agent, and a commercial light-cured composite. RESULTS: The study demonstrated that complete marginal adaptation was obtained by the experimental dentin bonding system used as the control. However, it was not possible to completely prevent gap formation with the commercial dentin bonding systems tested. A high correlation (R = 0.88) was observed between maximum contraction gap width and reduction of hardness only in the group in which the dentin was rinsed after cleaning. SIGNIFICANCE: It is very important to combine optimal dentin cleansers, primers, and bonding agents to compensate for the polymerization contraction stress of the composite. Therefore, it was speculated that the commercial dentin bonding systems tested in this study consisted of unsuitable materials. PMID- 7498601 TI - Dimensional changes related to ordering in an AuCu-3wt%Ga alloy at intraoral temperature. AB - OBJECTIVES: The purpose of this study was to evaluate the distortion behavior of a gold alloy with age-hardening ability at intraoral temperature induced by phase transformations. METHODS: Bridge-type specimens were prepared using AuCu-3wt%Ga alloy with age-hardening ability at 37 degrees C. Dimensional changes, hardness and surface roughness were measured. Transmission electron microscopy observation was carried out using AuCu-3wt%Ga discs aged at 37 degrees C for 30 d. RESULTS: Hardening was brought about rapidly in the initial stage of aging at 37 degrees C owing to ordering. The ordering was attributed to the formation of AuCu I and AuCu II superlattices. A linear contraction of 0.04% was induced by aging at 37 degrees C for 3 d. Distortion of the bridge-type specimen was brought about by a combination of linear contractions. The surface roughness increased in the initial stage of aging due to the surface relief effect of internal coherency strain caused by tetragonality. SIGNIFICANCE: The mode of distortion may be more advantageous for use in crown-type rather than inlay-type restorations, and the amount of contraction may not produce any problems in clinical use. PMID- 7498602 TI - Mold filling of Ti castings using investments with different gas permeability. AB - OBJECTIVES: The aim of this work was to study mold filling during casting of standardized five-unit Ti bridges vs. the gas permeability of the investments. METHODS: An Ar arc casting machine with a separate melting and mold chamber was used. Both chambers were evacuated before Ar was introduced into the melting chamber. A rubber sealer was inserted between the top of the steel ring of the mold and the steel floor between the two chambers. The mold cavity required the same Ar pressure as the melting chamber prior to casting. Four different investments for Ti casting were investigated: 1) Bellavest T (BEGO), 2) Rematitan Plus (Dentaurum), 3) Titavest CB (Morita) and 4) Titanium Vest (Ohara Co.). Three castings were made with each investment. The molds were fired according to the manufacturers' instructions and cooled to room temperature. The Ar gas pressures in the two chambers were recorded during casting, and the mold filling was evaluated initially by visual inspection and subsequently by studying the margins of the crowns in a microscope. RESULTS: In the melting and molding chambers, the Ar pressure was found to be: 1) Bellavest T: 400 and 1 torr; 2) Rematitan Plus: 600 and 85 torr; 3) Titavest CB:600 and 150 torr; and 4) Titanium Vest: 600 and 85 torr, respectively. This indicates substantial differences in the gas permeability of the molds. Bellavest T produced poorly filled molds, Rematitan Plus and Titanium Vest resulted in poor margins and Titavest CB produced complete castings. SIGNIFICANCE: Proper mold filling is promoted by investments with a high gas permeability, so the gases in the mold cavity can escape through the investment to the mold chamber. PMID- 7498604 TI - Primer for bonding resin to metal. AB - OBJECTIVES: The purpose was to examine the effect on the bond strength and durability of a resin bond to metal of modification of a primer consisting of thiophosphoric methacrylate with phosphoric methacrylates and/or benzoyl peroxide. METHODS: Acrylic rods were bonded with a luting resin consisting of poly(methyl methacrylate) powder and a mixture of methyl methacrylate and tributylborane initiator to silver-palladium alloy (Ag-Pd), gold-silver alloy (Au Ag), cobalt-chromium alloy (Co-Cr), and titanium (Ti) surfaces treated with various primers. The bonded samples were thermocycled for 2,000 cycles and the mean bond strengths were compared using one-way ANOVA and Duncan's new multiple range test at p < 0.05. RESULTS: Using primers of thiophosphoric methacrylate or phosphoric methacrylates alone, the bond strengths of Ag-Pd decreased significantly (p < 0.05) after thermocycling. The durability was significantly improved (p < 0.05) when thiophosphoric methocrylate was used in combination with the phosphate monomers. The additional use of benzoyl peroxide and heat treatment resulted in a significant increase (p < 0.05) in the durability of two groups; the mean bond strengths over 20 MPa and the lowest values remained unchanged even after 2,000 thermocycles. SIGNIFICANCE: The highest level of bond strength and durability to dental noble metals was achieved using a mixture of thiophosphoric and phosphoric methacrylates and benzoyl peroxide. These bond strength results are comparable to values obtained for base metals. PMID- 7498603 TI - Disinfection of dental stone casts: antimicrobial effects and physical property alterations. AB - OBJECTIVES: The purpose of this study was to evaluate the effectiveness of disinfecting solutions incorporated into dental stone casts against a standard and representative group of microorganisms and to note changes in the physical properties of the casts. METHODS: Irreversible hydrocolloid impressions were contaminated individually with Escherichia coli, Staphylococcus aureus, Enterobacter cloacae, Pseudomonas aeruginosa, Klebsiella pneumoniae, Actinobacter calcoaceticus, Bacillus subtilis, Mycobacterium phlei and Candida albicans. Four readily available disinfecting solutions (glutaraldehyde, povidone-iodine, chlorhexidine and sodium hypochlorite) were added to the die stone mix used to pour up the impressions. The set cast surfaces were swabbed at 1 h and 24 h, the samples plated on agar and incubated at 37 degrees C for 24 h and 3 d for M. phlei. Subsequently, colony forming units were counted. The physical properties assessed were setting time, setting expansion, compressive strength, detail reproduction and delayed expansion of the stone. RESULTS: Only glutaraldehyde and povidone-iodine killed all contaminating microorganisms within 1 h, while the 1:5 dilution of sodium hypochlorite solution was equally effective after 24 h. Two percent glutaraldehyde was the most effective disinfectant with the least adverse effects on the physical properties of the set cast. Although povidone-iodine caused a decrease in the compressive strength of the set cast, it can be considered to be a sound alternative. SIGNIFICANCE: This study supports the concept of incorporating disinfectants into model stone as a standard operating procedure for impressions of unknown history and, most sensibly, all dental impressions. PMID- 7498605 TI - Effect of zinc on strength and fatigue resistance of amalgam. AB - OBJECTIVES: This study was conducted to determine the effect of zinc in amalgam on the static mechanical properties and resistance to fatigue-crack propagation of amalgams. METHODS: Fatigue, creep, compressive and flexure tests were performed on high-Cu Dispersalloy (Johnson and Johnson Dental Products Co., East Windsor, NJ, USA) and low-Cu Velvalloy (S.S. White Dental Products Int., Philadelphia, PA, USA) in both Zn-containing and Zn-free formulations. Linear Elastic Fracture Mechanics principles were used to characterize the fatigue behavior (crack lengths were monitored). RESULTS: The incorporation of Zn into these amalgams significantly improved their fatigue and creep resistance, while the effect of Zn on the static compressive and flexure strengths was not consistent. Zn significantly increased (p < 0.05) the resistance to fatigue crack propagation during Stage II crack growth for both amalgams, and increased the variations in crack velocity for a given stress intensity difference, without visibly altering the path or nature of the fatigue cracks. Possible influences on fatigue behavior were the mixed microstructure (particles and matrix), the nature of the crack tip, and creep. SIGNIFICANCE: The superior resistance to tensile fatigue crack propagation of amalgams containing small amounts of Zn (approximately 1 wt%) in vitro compared with amalgams with no Zn correlated with the superior resistance to marginal breakdown in vivo of Zn-containing amalgams. The range of stress intensities over which stable cracks could propagate was small, while the large variations observed in fatigue crack growth rates for individual materials invalidate predicting a unique fatigue life from the empirical equations obtained. PMID- 7498606 TI - Time required to remove totally bonded tooth-colored posterior restorations and related tooth substance loss. AB - OBJECTIVES: The study was conducted to measure the time required to remove large totally bonded tooth-colored posterior restorations and related tooth substance loss. This information was collected to determine if there were differences between bonded restorations and conventional restorations. METHODS: Molars were restored with the following materials: amalgam, composite, glass ionomer cement or glass ceramic cusp coverages. After submitting them to an in vitro aging process, they were attached to a lower jaw model in a phantom head. Six dentists removed the test restorations under standardized, quasi-clinical conditions. RESULTS: While the amalgam was completely removed, some glass ionomer cement vestiges were found. In the glass-ceramic group, the margins were covered with remnants of the composite luting agent in several places. The most restorative material was left in the composite group. The loss of tooth structure after removal of amalgam, glass ionomer cement, composite and glass ceramic was 17.6 mm3, 19.6 mm3, 39.9 mm3 and 41.8 mm3, respectively. When comparing the removal time, the glass ionomer group scored best with an average of 11.9 min followed by the amalgam group with an average of 15.2 min, followed by 24.9 min for the composite group and 30.4 min for the glass ceramic group. SIGNIFICANCE: The removal of totally bonded posterior restorations made of composite and ceramic is more technically demanding and more time-consuming than the removal of glass ionomer and amalgam restorations. PMID- 7498607 TI - Corrosion of titanium and amalgam couples: effect of fluoride, area size, surface preparation and fabrication procedures. AB - OBJECTIVES: The aim of this investigation was to study the effect of surface treatments and electrode area size on the corrosion of cast and machined titanium in contact with conventional and high-copper amalgams in saline solutions with and without added fluoride ions. METHODS: The potentials and the charges transferred between amalgam and titanium couples were registered using standard electrochemical methods. RESULTS: Conventional amalgam corroded more than high copper amalgams in contact with titanium in saline solutions. Adding fluoride to the solution made the titanium potential more active and enhanced the corrosion of titanium in combination with high-copper amalgams. The amalgam corrosion increased with a five-fold enlargement of the titanium area. The increase was significant for one titanium-amalgam combination. Surface preparations affected the electrochemical behavior, and surface alterations were occasionally observed on wet-ground titanium specimens. No significant differences were found in comparisons of cast and lathe-cut titanium. SIGNIFICANCE: Surface preparations and fluoride affect the electrochemical activity of titanium. PMID- 7498608 TI - Thermocycling and dwell times in microleakage evaluation for bonded restorations. AB - OBJECTIVES: To answer criticisms of an insufficient number of cycles in thermocycling in an earlier paper (Wendt et al., 1992), this study was initiated to evaluate the effect of different dwell times during long-term thermocycling in microleakage analysis of bonded restoratives. METHODS: A total of 90 resin-bonded restorations, Tytin/Liner F, (Kerr/Bisco), APH/Universal Bond 3, (LD Caulk) and Heliomolar/Syntac, (Vivadent), were inserted in Class V preparations in human third molars. Group A restorations were stored in 0.5% basic fuchsin dye for 166 h. Group B restorations were thermocycled for 5000 cycles in 0.5% basic fuchsin dye baths at 5 +/- 1 degrees C and 55 +/- 1 degrees C with a dwell time of 10 s in each bath. Group C was thermocycled as in Group B but with a dwell time of 60 s. Readings by defined criteria were taken from each of two 1 mm sections along the cemental margin. RESULTS: There were no significant differences (p < 0.05) in the extent of dye penetration at the tooth restoration interface for the thermocycled composite restorations when compared to composite restorations with no thermocycling. There was a significant difference (p < 0.05) in the extent of dye penetration at the tooth restoration interface for the thermocycled amalgam restorations when compared to dye-exposed amalgam specimens with no thermocycling. SIGNIFICANCE: During microleakage analysis, the need for thermocycling is dependent upon the extent the restorative is thermally conductive in relation to its mass. The dwell time should be clinically relevant, i.e., 10 s, and is important only if the restorative is thermally conductive. PMID- 7498609 TI - Review of the 1994 dental materials literature. AB - OBJECTIVES: Electronic databases provide strong support for research and education literature searches. However, there is still some lag in time between publication and updates of the database. The objective of this summary was to identify and categorize the 1994 dental materials scientific literature that dealt with restorative dentistry. METHODS: Seventeen primary journals were manually searched via the Tables of Contents and 88 secondary journals were manually or electronically searched. Dental materials citations were categorized into 15 major sections and several subsections, excluding case reports and literature primarily related to dental implants or biomedical materials outside of dentistry. RESULTS: Nearly twelve hundred citations were identified, the largest number being associated with composites and glass ionomers. SIGNIFICANCE: This list provides a comprehensive source for review by academicians and researchers to bridge the gap between initial publication and electronic citation. PMID- 7498610 TI - Fatigue resistance of composite restorations: effect of filler content. AB - OBJECTIVES: The purpose of this study was to evaluate the effects of filler level on the fatigue impact resistance of resin composite. METHODS: A series of experimental composite materials was prepared by incorporating a silanized quartz filler (3-5 microns in size) into a light-cured resin matrix of Bis-GMA/TEGDMA. The filler contents in the experimental composites varied from 40 to 85 wt%. The composites were placed in standardized Class I cavities prepared in bovine teeth. The specimens were stressed with a repetitive impact load (1.6 x 10(2) joule) with loading cycles ranging from 50,000 to 150,000 times. The cracks induced by cyclic loading were observed on the sectioned surfaces of the tested specimens. RESULTS: The composites with considerably low or high filler content (< 60% or > 80% by weight) were significantly low in fatigue resistance. The results revealed that an inverse linear relationship tended to exist between filler level and fatigue resistance of the composite materials beyond a certain level of filler content. SIGNIFICANCE: Increased filler level does not necessarily improve the fatigue resistance of a resin composite as determined by applying a repetitive impact load. PMID- 7498611 TI - [Isolation of S. mbdakada from a child seen at the University of Messina pediatric clinic]. AB - The authors made some diagnostic investigation into a serious case of diarrhoea observed in an infant in the pediatric ward of the University hospital of Messina (Sicily). They brought into evidence some of the causes of the world wide spread of salmonella infection, in particular the spread of new serum-types, such as S. mbandaka, which was involved in this case. PMID- 7498612 TI - [A new serovar of Salmonella: S. IIIb 35: 1, z13: e, n, x, z15]. AB - The authors describe briefly the protocol for isolating a new serovar of Salmonella from the stools and the sand of a cage-shield of a python (Python regius (Shaw, 1802), kept as pet animal by a family in Messina. Such a finding is considered within the complex and wide investigation programme which we have been carrying out for years in our Institute of Veterinary Infectious Diseases, and within the problems rising from the Health and Hygiene field for the ever increasing diffusion of Enterobacteriaceae, by now limited to the ones collected and belonging to the Genus Salmonella. PMID- 7498613 TI - [Thoracic empyema caused by anaerobes. 78-month retrospective study. Clinical and microbiological aspects]. AB - The authors considered the microbiological and clinical aspects of the thoracic empyemas caused by anaerobic bacteria observed at the Ivrea-Castellamonte Hospital, Turin, over a period of 78 months. In this period 321 patients were subjected to thoracentesis in order to take samples of the pleural fluid (PF) which was in turn subjected to culture investigations. 31 patients (9.6%) had one or more cultures which were positive for bacterial growth. A good 12 patients (38.7%) had cultures which were positive for anaerobes alone or associated with aerobes. The average age of these patients (8 male, 4 female) was 62.5 years (range, 42 TO 84 Y.). Underlying diseases and predisposing conditions were as follows: malignancy (4 patients), neurological disorders (4p), severe hypoalbuminemia (5 p), bronchial obstruction (3 p), previous gastroenteric or chest surgery (3 p), diabetes mellitus (2 p), alcoholism (2 p), atherosclerosis (1 p), alveolar hypoventilation caused by chest wall disorder (1 p). The etiopathogenesis of the empyema was as follows; postpneumonic, 5 cases (41.7%); postsurgical, 4 cases (33.3%); esophageal fistula, 1 case (8.3%); idiopathic, 2 cases (16.7%). In 5 cases (441.7%) empyema developed in hospital, in 7 cases (58.3%) in community. Forty-one percent of the empyemas were described as foul smelling. The anaerobic bacteria most frequently isolated were Bacteroides spp, Fusobacterium nucleatum, Peptostreptococci in this order. Thoracentesis was performed on 6 patients, chest tube drainage was required in 6, and antibiotics were administered to all the patients. Length of Hospitalization averaged 33>2 days. Three patients (30%) died. These patients died during the same hospitalization period while empyema was an active problem.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7498614 TI - The role of antibiotics in the evolution of microorganisms. AB - Bacteria were the first living beings to appear on our planet: the most ancient fossils available, all of them were procaryotic microorganisms, developed 4, 5 billion years ago. The paleomicrobiological studies made on that kind of fossils, which are by now several hundreds in each continent, proved bacteria to have constantly evolved and to have originated the modern Eubacteria as well as the Archebacteria and the Cianobacteria. These last appeared about 2 billion years ago and, having acquired the oxygen-type photosynthesis, have caused the formation of a large amount of organic material, afterwards used by the younger organisms, and have modified the atmosphere introducing oxygen in it and conditioning in this way the other living being's evolution. From Bacteriaceae and Cyanobacteria derive the eucaryotic microorganisms (algae, fungi, protozoa, mould) and, little by little, all the other organisms both vegetable and animal subjected to the evolutionary pressures. Nevertheless bacteria undergo more frequently than all the others the evolution law because of their short reproductive time; this is the reason why bacteria are favourite compared with the other organisms. In fact each species is subjected to a genetic mutation every 10(5)/10(6) generations: in the vegetables and animals the consequences of a genetic mutation will be evident after millennia whereas in the bacteria the mutation happens in a short time. We ourselves have witnessed the revolution which took place in the bacteria populations during the last half century when numerous antibiotic-resistant strains appeared.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7498615 TI - [Antimicrobial activity of NK cells]. AB - NK Lymphocytes were first identified by their cytotoxic and cytolytic activity against tumor cells. However there is now increasing evidence that these cells are also mediators of natural resistance against viruses, fungi, protozoa and bacteria. The purpose of this review is to summarize the data on the antimicrobial activity of NK cells and to discuss the influence of corticoids and of other molecules interfering with innate resistance against pathogenic micro organisms. PMID- 7498616 TI - [Interferon alpha treatment in chronic hepatitis C]. AB - Alpha Interferon showed effective in the treatment of chronic type C hepatitis, but a consensus has not been reached about the selection of patients and therapy schedules so far. We treated 36 patients with chronic type C hepatitis in the outpatient ward of the 1st Infectious Diseases Dept., "Amedeo di Savoia" Hospital, Torino (Head of Dept.: Prof. W. Grillone) in the period 1990-1992. Alpha IFN 1-6 MU thrice weekly for 6-12 months was used. The average follow up period after therapy was 8.5 months. Four patients dropped out during the treatment period. The clinical response was evaluated using serum transaminases: 9 patients showed a full response, 9 patients had a hepatitis relapse after stopping the treatment, 6 patients had a partial response, and 9 were treatment failures. A better response was observed in young patients, drug addicts, with chronic persistent hepatitis and high transaminases levels. Side effects of the treatment were very frequent, but usually short lasting, and seldom responsible for dropping out. PMID- 7498617 TI - [Urinary infection. I. Comparative in vitro activities of several antibiotic drugs against Gram-negative bacteria]. AB - The in vitro activity of imipenem against 3,496 Gram-negative strains isolated from 4,505 positive urocultures was determined. The effectiveness of imipenem was then compared with that of ampicillin, amoxycillin + clavulanic acid, ticarcillin, piperacillin, aztreonam, cefalotin , ceftazidime, cefoxitin, cefotaxime, norfloxacin, ciprofloxacin, amikacin, gentamicin, tobramycin, trimethoprim + sulfamethoxazole. Imipenem was active against all isolates, including Pseudomonas spp. known to account for a large portion of nosocomial infections and resistance to antibacterial drugs. PMID- 7498618 TI - [Urinary infection. II. Comparative in vitro activities of several antibiotic drugs against Gram-positive bacteria]. AB - In this study the in vitro susceptibility of 1,009 Gram-positive strains isolated from 4,505 positive urocultures was evaluated towards the following chemotherapeutic agents: oxacillin, ampicillin, amoxicillin + clavulanic acid, imipenem, cefalotin, cefotaxime, amikacin, gentamicin, ciprofloxacin, pefloxacin, clindamycin, erythromycin, vancomycin, rifampicin, tetracycline and chloramphenicol. The results indicate that vancomycin is the most active antibiotic against all the strains, including enterococci often involved in serious infections caused by invasive techniques. PMID- 7498619 TI - [Planar myocardial scintigraphy with technetium-99m-sestamibi in acute myocardial infarct treated with thrombolytic therapy]. AB - BACKGROUND: Nuclear cardiology permits the estimation of the myocardial infarction size and the result of the thrombolytic therapy. The aim of the study was to demonstrate the feasibility of the planar myocardial scintigraphy with Technetium-99-m-sestamibi in the coronary intensive care unit for the early identification of the infarct size and the result of the thrombolytic therapy. MATERIALS AND METHODS: We considered 10 patients affected by a first myocardial infarction (5 anterior and 5 inferior wall) then treated with thrombolytic therapy (APSAC 30 U. iv) within an interval of 3 hours from the onset of the symptoms. Technetium-99-m-sestamibi was injected before the thrombolytic therapy and the planar imaging was registered after 2-3 hours with a mobile gamma-camera. After 24 hours and before patient discharge we repeated the scintigraphic evaluation. Within 24 hours from the thrombolytic therapy the coronary angiography was performed for the demonstration of patency of the infarct-related artery. The left ventricle myocardial perfusion was divided in the 3 planar projections into 13 segments. The perfusion in each segment was evaluated with a perfusion score: 0 = normal perfusion, 1 = moderately reduced, 2 = severely reduced, 3 = absent. The sum of the hypoperfused segments represented the infarct size. A perfusion score improvement greater than 40% was considered a marker of reperfusion. RESULTS: The infarct size involved 4.4 +/- 1.4 segments in the anterior and 2 +/- 0.6 segments in the inferior wall infarctions (p < 0.05). The scintigraphic imaging made 24 hours after the myocardial infarction allowed the diagnosis of coronary reperfusion in 7 patients. The coronary angiography demonstrated the infarct related artery patency in 9 patients (all with TIMI perfusion score = 3). The nuclear imaging at patient discharge provided the diagnosis or reperfusion in 8 cases and demonstrated an improvement of the myocardial perfusion score in 5 cases. CONCLUSION: The scintigraphic imaging with Technetium-99-m-sestamibi in the patients with a myocardial infarction treated with thrombolytic therapy is feasible with a mobile gamma-camera in the intensive coronary care unit. The quality of planar imaging is good and allows the evaluation of myocardial infarct size and efficiency of thrombolytic therapy. An earlier scintigraphic imaging should be taken into consideration for a more timely non-invasive evaluation of patients who need coronary angiography and, if necessary, a rescue PTCA. PMID- 7498620 TI - [The use of nuclear cardiology technics in assessing acute infarct in the era of thrombolysis: prospective or reality for cardiology?]. PMID- 7498621 TI - [Primary antiphospholipid syndrome with a familial element and myocardial infarct in an adolescent]. AB - We describe the case of a patient with primary familiar antiphospholipid syndrome and acute myocardial infarction. A previously healthy 15-year-old adolescent was admitted with severe chest pain lasting from 1 hour associated with inferoposterolateral ST-segment elevation. The patient received intravenous thrombolysis. A 2-dimensional echocardiogram revealed an area localized in the basal posterolateral left ventricular myocardium, that was akinetic and abnormally thin throughout the cardiac cycle. Peak creatinine kinase level was 1461 U/I. Subsequent electrocardiogram revealed inferoposterior infarction. Plasma anticardiolipin (aCL) IgG antibodies resulted positive (24 U.GPL) in repeated determinations. A dypiridamole echocardiographic test resulted negative. The patient's parents refused cardiac catheterization. He continues to do well at home 28 months after discharge. The patient's sister is affected by primary antiphospholipid syndrome characterized by recurrent abortion, very low platelet count and lupus anticoagulant positivity. Plasma aCL antibodies resulted positive also in the mother who did not have clinical manifestations. PMID- 7498622 TI - [Total atrioventricular block as the primary clinical manifestation of undiagnosed aortic dissection]. AB - We describe the case of a 54-year-old male who was examined due to light chest pain experienced the previous day. The man was admitted to the cardiology division for the presence of a complete atrioventricular block. On admission the echocardiogram showed a large aortic regurgitation not present two years earlier. Suspecting an aortic dissection we performed a transesophageal echocardiography and a contrast computerized tomography: the two examinations were negative. The next days the patient had two episodes of acute pulmonary edema so he was transferred to the regional reference hospital where an hemodynamic unit and cardiac surgery division were available. There, he repeated a transesophageal echocardiography which was negative. The man had aortic valve replacement without angiography for his very critical condition. The surgeon identified a small aortic dissection, just above the aortic valve plane, which was responsible for valve leaflets prolapse and aortic regurgitation; the hematoma deepened towards the interatrial septum and atrioventricular junction justifying the atrioventricular block. In conclusion, a small aortic dissection can offer an atypical picture, and in this condition even very valuable diagnostic techniques may fail the diagnosis. PMID- 7498623 TI - [Posttraumatic pericardial laceration and cleft of the anterior mitral leaflet]. AB - A 17-year-old patient sustained blunt trauma to the chest, during a motor vehicle accident, resulting in a pericardial laceration and cleft of anterior leaflet of mitral valve; both lesions were treated conservatively at a later date. The reasons of late symptomatology of valvular injury and absent initial identification are analyzed. Echocardiography's role for diagnosis, indication and choice of surgical treatment is emphasized. PMID- 7498624 TI - [The epidemiology of heart failure: a growing public health problem]. PMID- 7498625 TI - [Recent morphofunctional findings on the left atrium: their relations to thromboembolic risk]. AB - In the past, the left atrial appendage has been considered a "useless" structure but associated to thromboembolic complications; its physiologic role is still undefined. Owing to its great distensibility, left atrial appendage positively influences atrial compliance and left ventricular performances. In addition this structure seems to play an important role in circulatory homeostasis by the release of atrial natriuretic factor in response to volume loading and atrial stretch. Transesophageal echocardiography provides a detailed anatomical characterization of this structure and, by means of Doppler flow velocities recordings, supplies relevant functional data. Despite their anatomical contiguity, the left atrium and atrial appendage result from a separate embryonic development; likewise, their function may differentiate. In the left atrial appendage a quadriphasic flow pattern has been described in subjects with sinus rhythm; however, as we reported, in some patients a more complex Doppler pattern can be observed, with an additional systolic forward flow wave which is presumably due to reflection phenomena. In patients with atrial fibrillation, an irregular flow pattern has been detected, which is mostly dependent on the duration of the arrhythmia, the underlying heart disease and the left atrial pressure. By transesophageal echocardiography a clear correlation has been established between the left atrial appendage slow flow and the occurrence of thromboembolic events; however, while waiting data from large studies, stratification of patients according to thromboembolic risk and decisions about anticoagulant prophylaxis should be performed by clinical information and transthoracic echocardiographic findings. No clinical or echocardiographic parameter has been found to be predictive of the thromboembolic events after cardioversion; in this setting the exclusion of atrial or atrial appendage thrombi by transesophageal echocardiography does not rule out the need for anticoagulation in patients with atrial fibrillation undergoing electrical or pharmacological cardioversion. PMID- 7498626 TI - [Knowing how to read a scientific article. 3. Knowing how to assess the natural history of a disease]. PMID- 7498627 TI - [Chronic thromboembolic pulmonary hypertension: a surgical syndrome?]. PMID- 7498628 TI - [The organizational aspects and technics of catheter ablation in cardiac arrhythmias. Associazione Nazionale Medici Cardiologi Ospedalieri]. PMID- 7498629 TI - [A global evaluation of the quality of care in cardiac surgery]. AB - Until now, all reflections about cardiac surgery have been quantitative. Currently, quantitative saturation of cardiac surgery needs in France has been reached or even exceeded in some areas. Consequently, a qualitative approach becomes fundamental: to provide each patient, prior to the operation, with an assessment of his/her vital risk at a specific site; to allow cardiologists to objectively and statistically know the complete results obtained by various surgical teams; to incite improvements among surgical teams; and lastly, to allow the administration to assess services provided to patients. This study concerns 1271 patients consecutively operated on between January 1990 and December 1993 by the very same surgeon and represents the outline of a logical method of controlling clinical results. PMID- 7498630 TI - Normal behaviour of circulatory parameters during exercise. Reference values for heart rate and systemic blood pressure. The ECCIS Project data. Epidemiologia e Clinica della Cardiopatia Ischemica Silente. AB - The study of simultaneous variations in heart rate (HR) and systemic blood pressure is of great interest in ergometric practice complementing the analysis of the ST segment by ECG. This paper examines data proceeding from 500 consecutive, normal, exercise stress tests with the aim of offering reference values on the step-by-step behaviour of HR, systolic and diastolic blood pressure (SBP, DBP) during exercise in a normal population. The sample comes from a large epidemiological study (ECCIS Project) conducted on 4842 healthy, working men, aged 40-59, which proposes to identify, by a 3 stage procedure, subjects with totally asymptomatic coronary artery disease (type I silent ischemia). A further aim of our paper is to examine the influence of some physiological variables (age, height, weight, body mass index, resting HR, SBP and DBP) on the response to effort of HR, SBP and DBP; reciprocal HR/SBP adjustment during exercise; maximal attained workload and recovery time. Due to a preliminary observation that the rate of step-by-step increase in HR and SBP is inversely related to total duration, the population was split into 4 groups according to exercise tolerance (defined by maximal attained workload) to elaborate reference values. Furthermore our data demonstrate that: 1) SBP increases more rapidly with respect to HR for older and heavier subjects; 2) Exercise tolerance is inversely related to age, baseline HR and SBP, and directly related to weight and height; 3) return to baseline conditions, during recovery, is quicker for subjects with better exercise tolerance and lower baseline HR, SBP and weight. PMID- 7498631 TI - [The relationship between overweight, life style and casual and 24-hour pressures in a population of male subjects with mild hypertension. The results of the HARVEST study]. AB - AIM OF THE STUDY: To evaluate the relationship of coffee use, smoking, physical inactivity, alcohol intake and overweight with casual and ambulatory blood pressure in a large population of borderline to mild hypertensive subjects. METHODS: Six hundred and thirty men with a mean (+/- SE) age of 33 +/- 0.4 years (range = 18-45 years) and a mean office blood pressure of 146 +/- 0.4/94 +/- 0.2 mm Hg, participating in the multicentre HARVEST study, were divided into three categories according to: coffee consumption (0 cups, 1-3 cups, > 3 cups/day), number of cigarettes smoked per day (0, 1-10, 11-20), degree of physical activity (no activity, regular training, competitive activity), alcohol intake (no alcohol, < 50 g, > or = 50 g/day) and body mass index (tertiles). All patients underwent non invasive ambulatory blood pressure monitoring with either the A&D TM-2420 model 7 or the Spacelabs 90207 monitor. Moreover, 24-hour urine collection was made for epinephrine and norepinephrine assessment (n = 611). RESULTS: Twenty-four-hour systolic blood pressure was higher in the coffee drinkers than the nondrinkers (+2.6 mm Hg in the moderate drinkers). Instead, 24 hour diastolic blood pressure was mainly influenced by overweight (3.2 mm Hg difference between the low and high BMI tertiles) and physical inactivity (3.2 mm Hg difference between the sedentary men and the athletes). Generally, the association of the above factors was stronger with ambulatory than with office blood pressure, whereas alcohol intake was only related to office diastolic blood pressure. However, in a multivariate regression analysis alcohol use did not show an independent effect on either office or ambulatory blood pressure. Smoking showed a different effect on office and ambulatory blood pressure. In fact, office blood pressure was higher in the nonsmokers, while 24-hour blood pressure was higher in the smokers. Smoking, coffee and physical inactivity were associated with sympathetic overactivity, as documented by increased urinary catecholamines output. CONCLUSIONS: The results of the present study indicate that overweight and behavioural factors affect 24-hour blood pressure to a larger extent than office blood pressure does. This is likely to be due to their effect on the sympathetic nervous system activity, which would enhance the blood pressure response to daily life stressors. PMID- 7498632 TI - [Infections due to central catheterizations in a cardiac intensive care unit: an evaluation of 6 months of continuous surveillance. The nursing personnel of a cardiac intensive care unit]. AB - BACKGROUND: Infections in intensive care unit are nowadays well documented. As no recent papers about catheter related infections in cardiological intensive care units were found, we decided to approach them during a period of six months with a surveillance study including a clinical, nursing and microbiological protocol. METHODS: The microbiological protocol the semiquantitative Maki cultural evaluation of the tips, the interior of the hubs and the skin around the catheter insertion point. In addition, all the samples were additionally cultured in liquid media. A total of 432 biological samples were analysed, from 125 patients entered into our cardiological intensive care unit: 144 from catheter tips, 144 from the hubs and 144 from the skin around the catheter insertion. RESULTS: Three hundred and eighty-four biological samples (88.9%) were completely negative. Thirteen tips resulted positive (27.1% of the 48 total positive biological samples): 92.3% (12/13) of the positive catheters were infected from internal origin, hub and skin resulting culture negative. Of the 144 catheter tips 12 were colonized (C.F.U. < 15) while only 2 resulted infected (C.F.U. > or = 15). Only one patient was infected with an exogenous infection presenting tip, hub and skin positivity. The overall positivity was of 214 microorganisms mainly represented (93.5%) by gram positives; 87% of them were coagulase-negative staphylococci (C.N.S.). In addition, the tip detection of a Candida guillermondii strain in a pacemaker patient is a relevant finding. CONCLUSIONS: The very low number of cutaneous contaminations (1.3%) and patient's infections (0.69%) showed the good sensibilisation of the medical and nursing teams in the infection control and surveillance. PMID- 7498633 TI - [Urinary albumin excretion increases during an acute myocardial infarct especially in patients who develop heart failure]. AB - AIM OF THE STUDY: To evaluate the profile of albumin excretion rate (AER) in the first days of acute myocardial infarction (AMI), its relationship with serum enzymes and the presence of heart failure, and the effect of thrombolytic therapy. METHODS: Two hundred and thirty-one consecutive patients admitted to coronary care unit for suspected AMI were examined. Patients with diabetes mellitus, urinary tract infections or proteinuric diseases were excluded. In 135 patients (95 males, 40 females) AMI diagnosis was confirmed. The remaining 96 (56 males, 40 females) were considered as controls. AER was measured by radioimmunoassay in 24-hour urine samples at the first, third and seventh day after admission and expressed as mg/24h. Statistical analysis was performed after AER logarithmic transformation using repeated measure ANOVA: RESULTS: Mean age was 66.9 +/- 12.2 years (range = 35 -91) in the AMI group and 63.2 +/- 12.3 years (range = 33-91) in the controls (p = 0.023) Age-adjusted blood pressure was lower in the AMI group than in the controls (p < 0.0001 for both systolic and diastolic), while no difference was found in heart rate. Plasma cholesterol, triglycerides, creatinine and uric acid were similar in the 2 groups. Mean AER was 43.4 +/- 64.8, 26.9 +/- 51.2 and 23.9 +/- 52.7 mg/24h at 1st, 3rd and 7th day respectively in the AMI group and 24.9 +/- 58.2, 13.7 +/- 25.8 and 17.9 +/- 44.1 mg/24h respectively in the controls (p = 0.014). In the AMI group, first day AER significantly and positively correlated with CPK (r = 0.287, p = 0.001), CPK-MB (r = 0.239, p = 0.007) and GOT (r = 0.300, p = 0.001). Within the patients with AMI, those who developed heart failure (n = 57), had higher AER (48.6 +/- 68.4, 29.7 +/- 54.9 and 28.1 +/- 55.8 mg/24h at 1st, 3rd and 7th day in patients with mild heart failure -2nd Killip Class- and 100.0 +/- 141.7, 50.3 +/- 66.4 and 64.2 +/- 74.4 mg/24h in those with severe heart failure -3rd and 4th Killip Class-) than those who did not (31.0 +/- 41.7, 19.6 +/- 45.6 and 16.5 +/- 45.7 mg/24h respectively) (p = 0.004). In a multiple linear regression model AER was significantly related to peak values of GOT (1st day) and CPK (3rd day) and to presence of heart failure (3rd and 7th day). Thrombolytic therapy (n = 48) did not influence AER. CONCLUSIONS: The results of the present study show that AER increases following AMI, chiefly in the subjects who develop heart failure. AER correlates with serum enzymes peak levels at 1st and 3rd day and with presence of heart failure at 3rd and 7th day after admission, and is not influenced by thrombolytic therapy. These data suggest that in AMI the initial increase in AER is due to the inflammatory process which accompanies cardiac necrosis, while in a later phase its rise is mainly due to the increased intraglomerular capillary pressure consequent to heart failure. PMID- 7498634 TI - American Gastroenterological Association. American Gastroenterological Association directions: lessons from Chancellorsville. PMID- 7498635 TI - Presentation of the Julius M. Friedenwald Medal to John T. Farrar, M.D. PMID- 7498636 TI - Medical costs in community subjects with irritable bowel syndrome. AB - BACKGROUND & AIMS: Costs of management of irritable bowel syndrome (IBS) are unknown. The direct medical charges in community subjects with IBS were estimated. METHODS: An age- and sex-stratified random sample of residents of Olmsted County, Minnesota, ranging in age from 20 to 95 years, was mailed a valid self-report questionnaire. Subjects were categorized as having IBS, having some symptoms but inadequate criteria for IBS, and controls. All charges (in 1992 U.S. dollars) for health services rendered in the year before completing the survey were obtained (except outpatient medications). RESULTS: A total of 88% of subjects with IBS, 86% of subjects with some symptoms of IBS, and 83% of controls incurred direct medical charges during the study year. The odds of incurring charges were 1.6 times greater in subjects with IBS relative to those without symptoms (P < 0.01) adjusting for age, sex, education, marital status, and employment. Overall median charges incurred by subjects with IBS were $742 compared with $429 for controls and $614 for subjects with some symptoms. Among those subjects with nonzero charges, there were significant positive associations with age, higher education, and symptom groups (all P < 0.01) but not sex. CONCLUSIONS: The economic impact of IBS is significant. A better understanding of the determinants of these costs is needed so that cost-saving strategies can be implemented. PMID- 7498637 TI - Perception and reflex responses to intestinal distention in humans are modified by simultaneous or previous stimulation. AB - BACKGROUND & AIMS: Intestinal reflexes induced by distention in dogs are facilitated by either simultaneous or previous distentions. The aim of this study was to determine whether these phenomena also modulate the responses to intestinal distention, particularly perception, in humans. METHODS: Perception and intestinal relaxation were measured in 11 healthy subjects in response to increasing jejunal balloon distentions tested (by stimulus-response trials) alone, as control, and with conditioning distentions applied either simultaneously, immediately (10 seconds) before at the same site, or immediately before and 5 cm distant. In 8 additional subjects, the effect of prolonged (90 minute) conditioning distention was tested. RESULTS: Conditioning had more pronounced effects on perception than on intestinal reflexes. Perception of intestinal distention increased (by 84 +/- 47%; P < 0.05) when a simultaneous distention was applied nearby. By contrast, perception decreased (by 38 +/- 12%; P < 0.05) when a previous distention was applied at the same but not at an adjacent site. Prolonged intestinal distention elicited remarkably stable perception during a 90-minute period. The effects of conditioning were unrelated to intestinal compliance because it remained unchanged. CONCLUSIONS: In humans, temporospatial interactions of gut stimuli activate modulatory phenomena that determine the perception intensity of the stimuli. PMID- 7498638 TI - Changes in the site- and histology-specific incidence of gastric cancer during a 50-year period. AB - BACKGROUND & AIMS: In contrast to the dramatic decrease in the overall incidence of gastric cancer, there has been a reported increase in the incidence of cases located in the gastric cardia. The aim of this study was to identify changes in site- and histology-specific incidence rates of gastric adenocarcinoma during a 50-year period. METHODS: The Rochester Epidemiology Project medical records linkage system was used to identify all cases of gastric adenocarcinoma among residents of Olmsted County, Minnesota, between 1941 and 1990 (n = 342). Each patient's complete (inpatient and outpatient) medical records were reviewed and tumor location determined from pathological, surgical, endoscopic, and radiological reports. All available histological specimens (n = 246) were reviewed independently. RESULTS: The overall incidence of gastric cancer decreased from 48.8 per 100,000 person-years in the 1940s to 11.6 per 100,000 in the 1980s, whereas the incidence of adenocarcinoma of the cardia did not change significantly during the 50-year period. The incidence of adenocarcinoma of the esophagogastric junction increased from 0.0 to 1.9 per 100,000 person-years, but the number of cases was small. CONCLUSIONS: The incidence of adenocarcinoma of the gastric cardia has not increased in this population. The reported increase in cardia cancer in other populations may be due to an increasing incidence of adenocarcinoma of the esophagogastric junction. PMID- 7498639 TI - Esophageal body and lower esophageal sphincter function in healthy premature infants. AB - BACKGROUND & AIMS: Gastroesophageal reflux is a common problem in premature infants. The aim of this study was to use a novel manometric technique to measure esophageal body and lower esophageal sphincter pressures in premature infants. METHODS: Micromanometric feeding assemblies (OD, < or = 2 mm) incorporating 4-9 manometric channels were used in 49 studies of 27 premature neonates. Esophageal body motility was recorded at three sites for 20 minutes after feeding. Twenty attempts (one per minute) were made to stimulate swallowing via facial stimulation (Santmyer reflex). In 32 studies lower esophageal sphincter pressures were recorded (sleeve) for 15 minutes before and after feeding. RESULTS: Peristaltic motor patterns were less common than non-peristaltic motor patterns (26.6% vs. 73.4%; P < 0.0001) that comprised 31.1% synchronous, 34.6% incomplete, and 6.3% retrograde pressure waves. Reflex swallowing was elicited more frequently in neonates older than 34 weeks postconceptional age than in younger infants (33.4% vs. 20.4%; P < 0.05). Mean lower esophageal sphincter pressure was 20.5 +/- 1.7 mm Hg before and 13.7 +/- 1.3 mm Hg after feeding (P < 0.0005). CONCLUSIONS: Premature infants show nonperistaltic esophageal motility that may contribute to poor clearance of refluxed material. In contrast, the lower esophageal sphincter mechanisms seem well developed. PMID- 7498640 TI - Microsatellite instability in human colonic cancer is not a useful clinical indicator of familial colorectal cancer. AB - BACKGROUND & AIMS: Microsatellite instability is a property of most tumors occurring in the context of hereditary nonpolyposis colon cancer. Instability also occurs in 10%-15% of apparently sporadic colorectal cancers, and it has been hypothesized that this instability may indicate a genetic predisposition to colonic cancer. This study evaluated whether there is a clinically useful association between colon cancer instability and a family history of cancer. METHODS: Colon cancer cases (n = 188) from a population-based study were evaluated for microsatellite instability with 10 polymerase chain reaction primer sets. Instability results were compared with family history and other clinical and biological characteristics. RESULTS: Microsatellite instability was found in 16.5% of tumors. It was predominantly a feature of right-sided tumors (P = 0.003) and was associated with the youngest and oldest ages at diagnosis (P = 0.01). Instability was not associated with family history of cancer, sex of the individual, or the glutathione-S-transferase mu 1 null genotype. CONCLUSIONS: Although some very small, and as yet undefined, proportion of colon cancer may be caused by inherited mutations leading to microsatellite instability, tumoral instability by itself is not a marker for familiality and should not be considered as evidence for an inherited syndrome. PMID- 7498641 TI - Psychosensory modulation of colonic sensation in the human transverse and sigmoid colon. AB - BACKGROUND & AIMS: Psychosensory stimulation increases the perception of stimuli in different regions of the human colon. The aim of this study was to determine the perception of pain and gas during distention of two colonic regions in healthy volunteers during stress and relaxation. METHODS: In 22 healthy subjects, phasic distentions of transverse and sigmoid colon were performed using infinitely compliant balloons, and symptoms were measured before and during one of three randomized treatments: sham or active relaxation or mental stress. Anxiety ratings and somatic cold-pain sensitivity were included in a multivariate regression model. RESULTS: Mental stress increased levels of anxiety (P < 0.05) and the sensation of gas (P < 0.01), but not pain (P = NS), during transverse colon distentions and the sensations of gas and pain during sigmoid distentions (P < 0.05). Active relaxation reduced only sensation of gas in the sigmoid colon. Somatic pain sensitivity and pretreatment colonic sensory scores were significantly associated with the sensory scores during treatments. CONCLUSIONS: Psychosensory stimulation increases colonic sensation during short-term distention in two regions of the human colon; relaxation alone exerts a smaller influence on sensations arising in the sigmoid colon. PMID- 7498642 TI - Cost-effectiveness model for colon cancer screening. AB - BACKGROUND & AIMS: The relative efficacy and effectiveness of different colon screening programs has not been assessed. The purpose of this analysis was to provide a model for comparing several colon screening programs and to determine the key variables that impact program effectiveness. METHODS: Five screening programs were compared: annual fecal occult blood test (FOBT) alone, flexible sigmoidoscopy, flexible sigmoidoscopy and FOBT combined, one-time colonoscopy, and air-contrast barium enema. Key variables were adjusted for sensitivity analyses. Cost-effectiveness was defined as the cost per cancer death prevented. RESULTS: FOBT alone prevents fewer cancer deaths than the other programs. The addition of flexible sigmoidoscopy to the FOBT increases the rate of cancer prevention. One-time colonoscopy has the greatest impact on colorectal cancer mortality, largely because of assumptions that cancer would be prevented in most patients who undergo polypectomy. FOBT alone is the most cost-effective of the programs, but the cost is sensitive to several key variables. CONCLUSIONS: The model shows key variables that impact the cost-effectiveness of colon screening programs. Compliance is an important determinant of effectiveness of all of the screening programs. Future study should be focused on methods of patient education that improve patient compliance with screening. PMID- 7498643 TI - Coexpression of 5-HT2A and 5-HT4 receptors coupled to distinct signaling pathways in human intestinal muscle cells. AB - BACKGROUND & AIMS: The type and function of 5-hydroxytryptamine (5HT) receptors on intestinal muscle cells in humans are not known. 5-HT receptors were characterized pharmacologically and by radioligand binding. METHODS: Contraction, relaxation, inositol 1,4,5-triphosphate (IP3) and adenosine 3',5'-cyclic monophosphate (cAMP) formation, and 5-HT binding were measured in dispersed muscle cells and in cells in which only one receptor type was preserved by selective receptor protection. RESULTS: 5-HT binding was completely inhibited by 5-HT and partially by 5-HT2A (ketanserin), 5-HT4 (SDZ-205,557), and 5-HT1p (N acetyl-5-hydroxytryptophyl-5-hydroxytryptophan amide; 5-HTP-DP) receptor antagonists. 5-HT caused contraction that was inhibited by ketanserin and augmented by SDZ-205,557 and 5-HTP-DP. In the presence of ketanserin, 5-HT caused relaxation of cholecystokinin-contracted cells that was inhibited by SDZ-205,557 and 5-HTP-DP. 5-HT increased IP3, which was inhibited by ketanserin, and cAMP, which was inhibited by SDZ-205,557 and 5-HTP-DP. In cells with only 5-HT2A receptors, 5-HT caused contraction only, and residual binding was inhibited by ketanserin. In cells with only 5-HT4/5-HT1p receptors, 5-HT caused only relaxation and residual binding was inhibited by SDZ-205,557 and 5-HTP-DP. CONCLUSIONS: 5-HT2A receptors mediating contraction and 5-HT4 receptors mediating relaxation coexist on human intestinal muscle cells. The 5-HT4 receptors are closely similar or identical to 5-HT1p receptors. PMID- 7498644 TI - Management and outcome of patients with invasive carcinoma arising in colorectal polyps. AB - BACKGROUND & AIMS: Treatment for invasive adenocarcinoma in colorectal polyps (malignant polyps) is controversial. The aim of this study was to evaluate our institutional treatment strategy for malignant polyps. METHODS: Malignant polyps were designated as having favorable histology (grade I or II carcinoma with at least a 2-mm free margin) or unfavorable histology (grade III invasive adenocarcinoma, invasive adenocarcinoma with an unassessable margin, or a margin of < 2 mm). Malignant polyps with favorable histology were considered treated adequately by endoscopic polypectomy, whereas further therapy was recommended for malignant polyps with unfavorable histology. Recurrence, residual adenocarcinoma in a follow-up resection specimen, or metastasis during follow-up were considered adverse outcomes. RESULTS: Of the 47 patients identified, 17 (36%) had favorable histology. Sixteen patients (94%) were treated with polypectomy alone. None had an adverse outcome (median follow-up, 70 months). Thirty patients (64%) had unfavorable histology, and 21 patients (70%) underwent colectomy. Five patients underwent radiation therapy alone. Four patients underwent no additional therapy. Ten of 30 patients with unfavorable histology had adverse outcomes that differed significantly from the favorable histology group (P = 0.03). CONCLUSIONS: Endoscopic polypectomy alone is adequate therapy for malignant polyps with favorable histology. PMID- 7498645 TI - An intravenous loading dose of azathioprine decreases the time to response in patients with Crohn's disease. AB - BACKGROUND & AIMS: Azathioprine, an effective therapy for Crohn's disease, is limited by a prolonged time to response. The aim of this study was to determine the safety and utility of a loading dose of azathioprine to decrease the time to response in patients with Crohn's disease. METHODS: Twelve patients were studied: 6 with 13 fistulae and 6 with inflammatory disease. All patients received an intravenous infusion of azathioprine (50 mg/h for 36 hours). Response was determined by physical and radiographic examination for fistulae and by the Crohn's Disease Activity Index for inflammatory disease. Erythrocyte concentrations of azathioprine metabolites were measured by chromatography. RESULTS: Seven of 13 fistulae closed by week 4, and three had a temporary decrease in drainage. One fistula improved at week 16. Two fistulae failed to improve. Four of 6 patients with inflammatory disease achieved remission, and 1 improved temporarily. Improvement was rapid (< or = 4 weeks). Peak concentrations of azathioprine metabolites occurred within 3 days. Clinical response did not correlate with azathioprine metabolite concentrations at the azathioprine dose studied. No adverse events occurred. CONCLUSIONS: An 1800-mg intravenous loading dose of azathioprine is safe and may decrease the time to response to < or = 4 weeks in patients with Crohn's disease. Correlation between clinical response and azathioprine metabolite concentrations at larger azathioprine doses should be determined. PMID- 7498646 TI - Attenuation of esophageal shortening during peristalsis with hiatus hernia. AB - BACKGROUND & AIMS: Minimal quantitative information exists on esophageal shortening during peristalsis in the human esophagus. The aim of this study was to ascertain the effect of hiatus hernia on longitudinal muscle-mediated peristaltic esophageal shortening. METHODS: Seven volunteers and 11 patients with hiatal hernia had metal clips endoscopically affixed at the squamocolumnar junction and 3-5 cm proximal to it (n = 11). Location of the lower esophageal sphincter and axial clip movement were assessed using concurrent manometry and videofluoroscopy during barium swallows in a supine and upright posture with and without abdominal compression. RESULTS: Three subject groups were defined by the proximity of the squamocolumnar junction to the diaphragmatic hiatus: group 1, < or = 0 cm; group 2, between 0 and 2 cm; and group 3, > or = 2 cm. Peristaltic esophageal shortening was progressively diminished, re-elongation progressively prolonged, and the degree of contraction observed in the distal esophageal segment reduced with progressive degree of hiatus hernia. There was minimal mobility of the squamocolumnar junction relative to the hiatus with posture or abdominal compression. CONCLUSIONS: Longitudinal muscle contraction during peristalsis normally causes transient elevation of the squamocolumnar junction above the diaphragm. Esophageal shortening during primary peristalsis is reduced with increasing degree of hiatus hernia, suggesting that there is diminished opposition of longitudinal muscle contraction from the phrenoesophageal attachments. PMID- 7498647 TI - Platelet-activating factor acetylhydrolases in Caco-2 cells and epithelium of normal and ulcerative colitis patients. AB - BACKGROUND & AIMS: Platelet-activating factor (PAF) is a potent inflammatory mediator implicated in the pathogenesis of inflammatory bowel disease and necrotizing enterocolitis. Metabolism by platelet-activating factor acetylhydrolase (PAF-AH) is the major pathway for platelet-activating factor degradation. The aim of this study was to investigate the possible role of intestinal epithelium as a source of PAF-AH. METHODS: Intracellular and secreted PAF-AHs were characterized in human colonic epithelial cells isolated from histologically normal mucosa and inflamed mucosa from patients with ulcerative colitis and in the human intestinal epithelial cell line Caco-2 by measuring the metabolism of [3H]-PAF to [3H]lysoPAF. RESULTS: Human colonic epithelial cells and Caco-2 cells synthesize and secrete PAF-AH as shown by in vitro hydrolysis of [3H]PAF to [3H]-lysoPAF in cell lysates and conditioned media. Both intracellular and secreted PAF-AHs are calcium-independent and substrate-specific for phospholipids similar to PAF. Epithelial cells from involved areas of resections for ulcerative colitis had increased levels of secreted PAF-AH and decreased levels of intracellular PAF-AH compared with epithelial cells from histologically normal areas. CONCLUSIONS: Human colonic epithelial cells and Caco-2 cells produce intracellular and secreted PAF-AHs, which are distinct proteins. This is the first demonstration of PAF-AH production by epithelial cells. PMID- 7498649 TI - Shc is a substrate of the rat intestinal epidermal growth factor receptor tyrosine kinase. AB - BACKGROUND & AIMS: Epidermal growth factor (EGF) has been shown to induce intestinal proliferation and maturation; however, little information is available regarding substrates of the intestinal EGF receptor tyrosine kinase. The purpose of this study was to determine if src homologous collagen-like protein (Shc) was an in vivo substrate of the intestinal EGF receptor. METHODS: Ten-day-old rats were treated with EGF or were breast-fed. In some experiments, IEC-6 cells were treated with EGF. Intestinal tissue and cell fractions were studied by immunodetection to compare the tyrosine phosphorylation state and the subcellular localization of intestinal proteins. RESULTS: The total tyrosine phosphorylation state of intestinal proteins was increased threefold by EGF. Tyrosine phosphorylation of the EGF receptor and Shc were rapidly increased by EGF. The association of Grb2 with Shc increased fourfold and fivefold. Plasma membrane translocation of Shc and associated phosphotyrosyl proteins was increased within 30 seconds of EGF treatment. CONCLUSIONS: Shc is a substrate of the intestinal EGF receptor in vivo. EGF-induced association of Shc with the adapter protein Grb2 may have implications for activation of the p21ras signaling pathway in the intestine. The EGF-induced membrane association of Shc with two other phosphotyrosyl proteins suggests involvement of Shc in additional aspects of EGF receptor signaling in the intestine. PMID- 7498648 TI - The site-specific delivery of ursodeoxycholic acid to the rat colon by sulfate conjugation. AB - BACKGROUND & AIMS: Because ursodeoxycholate has been shown to act as a tumor suppressive agent in the colon, the absorption and metabolism of its sulfate conjugates were examined in rats to show that sulfation would facilitate the site specific delivery of ursodeoxycholate to the colon. METHODS: Bile acids were measured in intestinal contents, feces, urine, plasma, and liver tissue after oral administration of ursodeoxycholate and its C-3, C-7, and C-3,7 sulfate derivatives. RESULTS: Ursodeoxycholate was found in the jejunum after administration of all bile acids, but the mass was greatest for ursodeoxycholic acid administration. In the colon, lithocholic acid, normally found in negligible amounts, became the major bile acid after ursodeoxycholate administration. In contrast, reductions in mass and proportions of lithocholate and deoxycholate occurred after administering the C-7 sulfates. The fecal lithocholate/deoxycholate ratio, a risk marker for colon cancer, increased markedly after administration of ursodeoxycholate and its C-3 sulfate, but did not change after administering the C-7 sulfates. Unlike ursodeoxycholate or its C 3 sulfate, which increased liver concentrations of lithocholate and ursodeoxycholate, the C-7 sulfates had the opposite effect, which was consistent with poor absorption. CONCLUSIONS: Sulfation of ursodeoxycholate, specifically at the C-7 position, protects the molecule from bacterial degradation and inhibits its intestinal absorption, thereby facilitating delivery to the colon. PMID- 7498650 TI - Involvement of eicosanoids and macrophage-like cells in cytokine-mediated changes in rat myenteric nerves. AB - BACKGROUND & AIMS: Proinflammatory cytokines alter function in enteric nerves, but little is known about underlying mechanisms. This study was designed to investigate the roles of prostanoids and of macrophage-like cells in cytokine induced suppression of [3H]norepinephrine release from rat myenteric plexus. METHODS: The release of 3H from jejunal longitudinal muscle-myenteric plexus preparations that had been loaded with [3H]norepinephrine was measured. Measurements of 3H release as well as concentrations of prostaglandin E2 and leukotriene were made in preparations exposed to interleukin 1 beta plus interleukin 6 and in the presence or absence of piroxicam, 5-lipoxygenase inhibitor MK886, cycloheximide, or cyclosporin A. An ultrastructural analysis was also performed to investigate the presence of macrophage-like cells in the myenteric plexus. RESULTS: Interleukin 1 beta plus interleukin 6 suppressed 3H release and caused an increase in tissue prostaglandin E2 but not leukotriene E4. Piroxicam and cycloheximide but not MK886 attenuated the cytokine-induced increase in prostaglandin E2 and the suppression of [3H]norepinephrine release. Ultrastructural analysis showed macrophage-like cells in the plexus, and the cytokine effects were inhibited by cyclosporin A. CONCLUSIONS: Prostanoids but not leukotrienes mediate the cytokine-induced suppression of norepinephrine release, and the results of this study suggest that macrophage-like cells are also involved. PMID- 7498651 TI - Guanylin, an endogenous ligand for C-type guanylate cyclase, is produced by goblet cells in the rat intestine. AB - BACKGROUND & AIMS: Guanylin activates an intestinal guanylate cyclase (GCC) and stimulates electrolyte movement across the gut epithelium. Cells expressing guanylin messenger RNA have been localized to the epithelial cell layer of the intestine; however, the identity of the guanylin-producing cells has not been determined. The aim of this study was to identify cells that express guanylin in the rat intestine. METHODS: Antibodies were raised against defined proguanylin epitopes, evaluated by Western blotting, and used for immunoperoxidase histochemistry. RESULTS: Guanylin-like immunoreactivity was localized to a subset of goblet cells. In the small intestine, most, perhaps all, goblet cells in the villi were immunopositive, as were some goblet cells in upper crypts; however, goblet cells deep within crypts were unlabeled. In the colon, goblet cells clustered in the necks and around the openings of crypts were immunopositive, whereas (as in the small intestine) goblet cells in deeper crypt regions were unlabeled. In some animals, immunoreactive columnar epithelial cells were also observed in the colon (although such cells were not apparent in the small intestine). Relative labeling of columnar cells varied from animal to animal. CONCLUSIONS: Guanylin is expressed in mature goblet cells. If secreted in conjunction with mucin, it could play a role in the hydration of mucus. PMID- 7498652 TI - Absence of human papillomavirus DNA detected by polymerase chain reaction in French patients with esophageal carcinoma. AB - BACKGROUND & AIMS: Recent studies have suggested that esophageal human papillomavirus infection could be a risk factor for esophageal squamous cell carcinoma. The aim of this study was to evaluate the prevalence of human papillomavirus DNA sequences in the esophagus of French patients with esophageal squamous cell carcinoma. METHODS: Multiplex polymerase chain reactions with consensus primers directed to the L1 gene or specific primers for human papillomavirus types 6, 11, 16, 18, 31, and 33 directed to E6 gene (40 cycles followed by restriction mapping of the amplified products) were used to determine the presence of human papillomavirus DNA sequences in esophageal squamous cell carcinoma (n = 75), normal adjacent mucosa (n = 49), and metastatic lymphadenopathies (n = 5). As an internal control, a target located in the embryonic myosin heavy-chain gene was used in each reaction. RESULTS: Human papillomavirus DNA sequences could not be detected in any of the tumoral samples, the normal adjacent mucosa, or the metastatic lymphadenopathies. CONCLUSIONS: Human papillomavirus seems not to be implicated in esophageal carcinogenesis, at least in French patients, because the viral genomes are not associated with esophageal squamous cell carcinomas. PMID- 7498653 TI - Interruption of a transforming growth factor alpha autocrine loop in Caco-2 cells by antisense oligodeoxynucleotides. AB - BACKGROUND & AIMS: We have previously shown that Caco-2 cell proliferation is driven by basolateral membrane epidermal growth factor receptors. The aim of this study was to investigate whether autocrine production of transforming growth factor alpha (TGF-alpha) activates these receptors and stimulates proliferation using antisense oligodeoxynucleotides. METHODS: Caco-2 cells grown on microporous membranes or Jurkat cells were exposed to conventional or 5' cholesterol-modified oligodeoxynucleotides synthesized with random, antisense, or missense base sequences. Indices of proliferation were measured, including [3H]thymidine or [3H]uridine uptake for studies of short-term stimulation and the methylthiotetrazole assay as an index of cell number increase over longer periods. Secretion of TGF-alpha by cells was detected using a soft agar bioassay. RESULTS: Incubation with antisense oligodeoxynucleotides inhibited TGF-alpha secretion compared with controls. Random and missense oligodeoxynucleotides had no effect on proliferation. The TGF-alpha antisense oligodeoxynucleotides markedly inhibited proliferation, an effect that was abolished by adding TGF alpha to the medium. Oligonucleotides had no effect on Jurkat cells, a lymphocytic cell line lacking epidermal growth factor receptors. Cholesterol modified oligodeoxynucleotides were more effective and specific than unmodified oligodeoxynucleotides. CONCLUSIONS: Caco-2 cell proliferation is driven by autocrine stimulation of epidermal growth factor receptors by TGF-alpha. This mechanism may be effectively inhibited by antisense oligodeoxynucleotides, particularly those modified by the 5' attachment of cholesterol. PMID- 7498654 TI - Effect of germfree state on the capacities of isolated rat colonocytes to metabolize n-butyrate, glucose, and glutamine. AB - BACKGROUND & AIMS: Among substrates available to the colonic mucosa, n-butyrate from bacterial origin represents a major fuel. The present work investigated possible modifications of energy substrate metabolism in colonocytes isolated from germfree rats. METHODS: Colonocytes isolated from germfree vs. conventional rats were incubated (30 minutes at 37 degrees C) in the presence of 14C-labeled n butyrate (10 mmol/L), glucose (5 mmol/L), or glutamine (5 mmol/L). 14CO2 and metabolites generated were measured. Possible regulatory steps were also investigated. RESULTS: Glucose use rate was 25% lower in germfree rat colonocytes due to a reduced glycolytic capacity in these cells. Differences in 6 phosphofructo-1-kinase activity could account for this decrease. In contrast, glutamine use rate was 45% higher, and this was correlated with a higher maximum velocity of glutaminase in these cells. Nevertheless, the capacities to oxidize glucose and glutamine remained unchanged. Although the capacity to use n-butyrate was maintained in colonocytes of germfree rats, the ketogenic capacity was lower, whereas the capacity to oxidize n-butyrate was higher. The mitochondrial 3 hydroxy-3-methylglutaryl-coenzyme A synthase protein was identified in the colonic mucosa. Moreover, the messenger RNA and amount of protein were 75% lower in the germfree state. CONCLUSIONS: The absence of an intestinal microflora induces specific changes in the metabolic capacities of colonocytes. PMID- 7498655 TI - Hansenula anomala killer toxin induces secretion and severe acute injury in the rat intestine. AB - BACKGROUND & AIMS: The yeast Hansenula anomala has been associated with gastrointestinal symptomatology and damage to the intestinal wall in humans. In vitro and in vivo, H. anomala secretes a toxin, killer toxin, which is lethal to other microorganisms. In view of the very high rate of killer phenotype expression recorded for H. anomala strains in nature, this study aimed to investigate the hypothesis that H. anomala killer toxin plays a role in the pathogenesis of H. anomala-induced enteritis. METHODS: Effects of active and heat inactivated H. anomala killer toxin on intestinal fluid homeostasis and electrolyte balance were investigated in rat small intestine using a standard intestinal perfusion technique. Sections of the perfused jejunum tracts were examined histologically. RESULTS: H. anomala killer toxin induced a significant secretion of water and electrolytes. No significant change was observed when either heat-inactivated H. anomala killer toxin or control growth medium were tested. Histological analysis showed ischemic degeneration of villi and sloughing of surface epithelium in 50% of active H. anomala killer toxin-perfused jejuna. CONCLUSIONS: This paper presents original observations compatible with the hypothesis that H. anomala killer toxin plays a role in the pathogenesis of H. anomala-induced enteritis. PMID- 7498656 TI - A 7-year experience of severe acetaminophen-induced hepatotoxicity (1987-1993). AB - BACKGROUND & AIMS: Five hundred sixty patients admitted between January 1, 1987, and December 31, 1993, with severe acetaminophen-induced hepatotoxicity were studied. The aim of this study was to identify why severe acetaminophen-induced hepatotoxicity still occurs and to determine how known risk factors and advances in management have affected the pattern of illness and outcome. METHODS: This was a retrospective study of the etiologic factors and the clinical course of all acetaminophen-related admissions. RESULTS: The number of admissions increased from 58 in 1987 to 123 in 1993. During the corresponding period, overall survival improved from just < 50% to 78%. The percentage of admissions treated with N acetylcysteine increased from 40% in 1987 to 83% in 1993. The frequency with which grade III or IV encephalopathy developed decreased from 62% in 1987 to 40% in 1993, and the percentage of these patients who developed cerebral edema decreased from 61% to 45% during the same period. There was an increase in both the number of patients transplanted and the survival of those managed medically. CONCLUSIONS: Severe acetaminophen-induced hepatotoxicity remains a serious condition, but the increasing use of N-acetylcysteine, advances in medical management, and the increasing availability of transplantation have resulted in a significant improvement in survival rates. PMID- 7498657 TI - Effect of volume expansion on systemic hemodynamics and central and arterial blood volume in cirrhosis. AB - BACKGROUND & AIMS: Systemic vasodilatation in cirrhosis may lead to hemodynamic alterations with reduced effective blood volume and decreased arterial blood pressure. This study investigates the response of acute volume expansion on hemodynamics and regional blood volumes in patients with cirrhosis and in controls. METHODS: Thirty-nine patients with cirrhosis (12 patients with Child Turcotte class A, 14 with class B, and 13 with class C) and 6 controls were studied. During hepatic vein catheterization, cardiac output, systemic vascular resistance, central and arterial blood volume, noncentral blood volume, and arterial pressure were determined before and during a volume expansion induced by infusion of a hyperosmotic galactose solution. RESULTS: During volume expansion, the central and arterial blood volume increased significantly in patients with class A and controls, whereas no significant change was found in patients with either class B or class C. Conversely, the noncentral blood volume increased in patients with class B and C. In both patients and controls, the cardiac output increased and the systemic vascular resistance decreased, whereas the mean arterial blood pressure did not change significantly. CONCLUSIONS: Only in mild cirrhosis is the effective blood volume able to increase in response to volume expansion. Our results are consistent with the peripheral vasodilatation hypothesis and the circulatory hyporeactivity occurring in advanced cirrhosis. PMID- 7498659 TI - Virion-like structures in HeLa G cells transfected with the full-length sequence of the hepatitis C virus genome. AB - BACKGROUND & AIMS: The process and the site of hepatitis C virus (HCV) particle formation in cells after infection remain unknown. The aim of this study was to create an in vitro model for the study of HCV particle formation. METHODS: HeLa G cells were transfected with the full-length sequence of the HCV genome. Viral protein expression was analyzed using immunoblotting. The cells were examined using immunoelectron and conventional electron microscopy. RESULTS: Core, E2, NS3, NS5a, and NS5b proteins were identified using immunoblotting. Immunoelectron microscopy showed that the core antigen was located along the membrane of the endoplasmic reticulum (ER) and occasionally in its cisternae. Core antigen positive particles of 30 nm in diameter were found in the cytosol and in the cisternae of the ER. The particles in the cisternae were coated with an outer membrane that was connected to the ER membrane. Conventional electron microscopy revealed particles of 45 nm in diameter with electron-dense cores in the cisternae of the ER. The outer membrane of the particles was occasionally connected to the ER membrane. CONCLUSIONS: The findings suggest that HCV core proteins are synthesized and assembled into particles in the cytosol and that they bud into the cisternae of the ER to form coated particles. PMID- 7498660 TI - Antioxidant activity of silybin in vivo during long-term iron overload in rats. AB - BACKGROUND & AIMS: Hepatic iron toxicity may be mediated by free radical species and lipid peroxidation of biological membranes. The antioxidant property of silybin, a main constituent of natural flavonoids, was investigated in vivo during experimental iron overload. METHODS: Rats were fed a 2.5% carbonyl-iron diet and 100 mg.kg body wt-1.day-1 silybin for 4 months and were assayed for accumulation of hepatic lipid peroxidation by-products by immunocytochemistry, mitochondrial energy-dependent functions, and mitochondrial malondialdehyde content. RESULTS: Iron overload caused a dramatic accumulation of malondialdehyde protein adducts into iron-filled periportal hepatocytes that was decreased appreciably by silybin treatment. The same beneficial effect of silybin was found on the iron-induced accumulation of malondialdehyde in mitochondria. As to the liver functional efficiency, mitochondrial energy wasting and tissue adenosine triphosphate depletion induced by iron overload were successfully counteracted by silybin. CONCLUSIONS: Oral administration of silybin protects against iron induced hepatic toxicity in vivo. This effect seems to be caused by the prominent antioxidant activity of this compound. PMID- 7498661 TI - Antibody-targeted lymphokine-activated killer cells inhibit liver micrometastases in severe combined immunodeficient mice. AB - BACKGROUND & AIMS: Animal models for hepatic metastases can facilitate the investigation of lymphokine-activated killer (LAK) cell-based immunotherapy. The aim of this study was to investigate the efficacy of ccM4 antibody-targeted LAK cells in inhibiting hepatic micrometastases. METHODS: Hepatic micrometastases were generated after the intrasplenic injection of HM7 colon carcinoma cells. TAG72 expression was detected in these hepatic micrometastases using ccM4 antibody. The ccM4 antibody was conjugated onto LAK cells by treatment with 17.5% polyethylene glycol 8000. After the intrasplenic injection of HM7 cells, severe combined immunodeficient mice were randomized into five groups (i-v) and received either 10(7) ccM4-LAK cells plus 1000 U interleukin 2 (IL-2; group i), LAK cells plus 50 micrograms ccM4 and IL-2 (group ii), LAK cells plus IL-2 (group iii), IL 2 alone (group iv), or only phosphate-buffered saline (group v). RESULTS: The ccM4-LAK cells retained cytolytic activity and acquired TAG72-binding reactivity. The results showed that group i had significantly fewer hepatic metastases compared with group ii or group iii (P < 0.05) and even fewer hepatic metastases compared with group iv or group v (P < 0.001). CONCLUSIONS: These results show that ccM4 antibody-targeted LAK cells significantly inhibited tumor growth in vivo; thus, they can be potentially useful in treatment of hepatic micrometastases. PMID- 7498658 TI - Subcellular localization of hepatitis B core antigen in relation to hepatocyte regeneration in chronic hepatitis B. AB - BACKGROUND & AIMS: To test whether the dominant cytoplasmic expression of hepatitis B core antigen (HBcAg) in active chronic hepatitis B is secondary to liver damage and regeneration, the relationship between subcellular localization of HBcAg, liver inflammatory activity, and hepatocyte regeneration in chronic hepatitis B was studied. METHODS: Correlation of the clinical and laboratory data with the topographical distribution of HBcAg was studied in 30 patients. The subcellular localization of HBcAg in relation to hepatocyte cell cycles was studied by double immunostaining of HBcAg and proliferating cell nuclear antigen. RESULTS: Patients with predominant cytoplasmic HBcAg had significantly higher levels of biochemical and histological activities and proliferating cell nuclear antigen expression than patients with predominant nuclear HBcAg. The levels of proliferating cell nuclear antigen expression correlated positively with biochemical and histological activities and degrees of cytoplasmic HBcAg expression but negatively with degrees of nuclear HbcAg expression. Proliferating cell nuclear antigen expression was shown in 49% of hepatocytes with cytoplasmic HBcAg but in only 2% of hepatocytes with nuclear HBcAg. CONCLUSIONS: These findings suggested that, following liver damage, the regeneration of surviving hepatocytes might cause the shift of intracellular HBcAg from nucleus to cytoplasm. As a result, the extent of nuclear HBcAg expression reduces with concomitant increase in cytoplasmic HBcAg expression. PMID- 7498662 TI - Rat Kupffer cell-derived nitric oxide modulates induction of lymphokine-activated killer cell. AB - BACKGROUND & AIMS: Nitric oxide is now recognized to regulate immune responses and cell viability in various organs. The present study was designed to clarify whether NO released from Kupffer cells modulates the lymphokine-activated killer (LAK) activity of interleukin 2 (IL-2)-treated splenocytes. METHODS: Splenocytes and Kupffer cells were isolated from male Wistar rats and cocultured for 48 hours in the presence of lipopolysaccharide (1 microgram/mL). The splenocyte LAK activity and expression of IL-2 receptor were determined. RESULTS: Kupffer cells with lipopolysaccharide reduced the IL-2 receptor expression and LAK activity of splenocytes. The addition of either NG-monomethyl-L-arginine, an inhibitor of NO synthesis, or aminoguanidine, an inhibitor of inducible NO synthase, to the medium reversed the suppression of IL-2 receptor expression and LAK activity by lipopolysaccharide-stimulated Kupffer cells. 8-bromoguanosine 3',5'-cyclic monophosphate and NO donors decreased the splenocyte LAK activity and IL-2 receptor expression. Treatment with lipopolysaccharide increased the inducible NO synthase activity as well as the nitrite and nitrate levels in the culture medium of Kupffer cells but not in splenocytes. CONCLUSIONS: The results of this study suggest that NO produced by the inducible NO synthase of Kupffer cells in response to lipopolysaccharide modulates the IL-2 receptor expression and LAK activity of splenocytes. PMID- 7498663 TI - Role of hepatocytes in direct clearance of lipopolysaccharide in rats. AB - BACKGROUND & AIMS: The liver is the clearance organ for lipopolysaccharide (LPS). The aim of this study was to investigate the biliary excretion of LPS using fluorescein isothiocyanate (FITC)-labeled LPS. METHODS: After FITC-LPS was injected intravenously into rats, the cellular localization of fluorescence in the liver was examined and the biliary excretion of fluorescence was measured. The effects of gadolinium chloride, a blocker of Kupffer cells, and colchicine, an inhibitor of microtubules, on the biliary excretion of fluorescence was investigated, and bile was analyzed using high-performance liquid chromatography. RESULTS: Laser scanning confocal microscopy showed that fluorescence was taken up by hepatocytes 5 minutes after injection of FITC-LPS into the portal vein. When FITC-LPS was injected into the portal vein, fluorescence was rapidly secreted into bile, peaking at 20 minutes, and 25.1% of the injected dose appeared in bile within 60 minutes. When the same dose of FITC-LPS was injected into the tail vein, 15.8% appeared in bile within 60 minutes. Chromatography showed that FITC LPS was excreted into bile in an unchanged form over a period of 20 minutes after injection. Colchicine significantly reduced the biliary excretion of fluorescence, but gadolinium chloride had no effect. CONCLUSIONS: LPS was directly and effectively processed by hepatocytes and secreted into the bile canalicular system via a microtubule-dependent vesicular pathway. PMID- 7498665 TI - Fibroblast growth factor 2 and transforming growth factor beta 1 interactions in human liver myofibroblasts. AB - BACKGROUND & AIMS: During liver fibrogenesis, myofibroblastic liver cells proliferate and synthesize components of fibrosis. Fibroblast growth factor 2 (FGF-2) is expressed in vivo in myofibroblastic liver cells (MFLCs) during fibrogenesis, and exogenous FGF-2 is mitogenic for MFLCs. The aim of this study was to study the expression and role of endogenous FGF-2 in cultured human MFLCs. METHODS: FGF-2 and FGF-2 receptors were studied using immunoblotting. All RNA studies used ribonuclease protection. Growth of MFLCs was studied using [3H]thymidine incorporation and direct cell counting. RESULTS: MFLCs expressed FGF-2 and its receptors FGF receptor 1 and FGF receptor 2. An antibody to FGF-2 blocked the mitogenic effect of transforming growth factor beta 1 (TGF-beta 1) for MFLCs but not TGF-beta 1-induced increase in cellular fibronectin messenger RNA (mRNA). TGF-beta 1 increased levels of FGF-2 and FGF receptor mRNAs in MFLCs. We have previously shown that TGF-beta 1 also increased platelet-derived growth factor (PDGF) A chain mRNA in these cells and that anti-PDGF antibody blunted the mitogenic effect of TGF-beta 1. The present results show that anti-FGF-2 and anti PDGF-AA are not additive and that FGF-2 and PDGF-AA are not sequentially induced by TGF-beta 1. CONCLUSIONS: FGF-2 mediates the mitogenic but not the profibrogenic effect of TGF-beta 1 for human MFLCs, and autocrine FGF-2 and PDGF A interact in the mediation of the mitogenic effect of TGF-beta 1. PMID- 7498667 TI - Fraternal sisters with adult polycystic kidney disease and adenoma of the ampulla of Vater. AB - The cases of two fraternal sisters with symptomatic biliary obstruction due to adenomas of the ampulla of Vater are reported. Both sisters had autosomal dominant adult polycystic kidney disease. There are no previous reports of a familial occurrence of ampullary adenomas in the absence of familial adenomatous polyposis, nor has an association between autosomal dominant polycystic kidney disease and ampullary adenoma been described. The coexistence of both disorders in these sisters raises the possibility of a genetic link between autosomal dominant polycystic kidney disease and ampullary adenoma. PMID- 7498666 TI - Regulation of protein secretion into bile: studies in mice with a disrupted mdr2 p-glycoprotein gene. AB - BACKGROUND & AIMS: Protein is secreted into bile via several independent pathways. The aim of this study was to investigate whether these pathways are influenced by secretion of biliary lipid. METHODS: Protein secretion and biliary lipid output were studied in wild-type mice (+/+), heterozygotes (+/-), and homozygotes (-/-) for mdr2 gene disruption. Biliary lipid and protein output were varied by infusion with taurocholate (TC) and tauroursodeoxycholate (TUDC). RESULTS: Exocytosis and transcytosis were unaltered in (-/-) mice. Infusion with TC strongly induced secretion of alkaline phosphatase in (-/-) mice but had little effect in (+/-) and (+/+) mice. Infusion with TUDC had little effect on alkaline phosphatase output. In contrast, both TUDC and TC strongly stimulated secretion of aminopeptidase N and lysosomal enzymes in (+/+) mice but had no effect in (-/-) animals. Aminopeptidase N secretion correlated with phospholipid output, but only at high flux. At low flux, aminopeptidase N was secreted independently from both phospholipid and bile salts. CONCLUSIONS: The canalicular membrane enzymes alkaline phosphatase and aminopeptidase N are secreted via separate pathways. Part of alkaline phosphatase output is controlled by bile salt hydrophobicity, whereas at high lipid flux, aminopeptidase N secretion seems to be coupled to phospholipid output. Lysosomal enzymes follow the latter pathway. PMID- 7498664 TI - Intraileal carbohydrate regulates canine postprandial pancreaticobiliary secretion and upper gut motility. AB - BACKGROUND & AIMS: The effect of nutrients in the distal small intestine or colon on postprandial upper gut function is incompletely understood. The aim of this study was to determine if carbohydrate in the ileum or proximal colon of dogs affects postprandial pancreaticobiliary secretion, gastrointestinal transit, and circulating concentrations of certain gastrointestinal regulatory peptides. METHODS: Seven dogs were prepared with permanent infusion and aspiration catheters in the duodenum and ileum and an infusion catheter in the cecum. Coincident with eating a meal containing liquid and solid markers, ileal or colonic (n = 5 dogs for each) infusion were begun of isosmolar 0.9% NaCl or carbohydrate in a 3:1 ratio of starch to glucose. Pancreatic enzyme output, bile acid delivery, gastrointestinal polypeptide, and plasma concentrations of pancreatic polypeptide, neurotensin, and peptide YY were measured for 6 hours postprandially. RESULTS: Carbohydrate infusion in the ileum, but not in the proximal colon, increased amylase secretion and plasma peptide YY, slowed gastric emptying of liquids and solids, slowed small intestinal transit, and decreased bile acid delivery into the duodenum (P < 0.05 in each). CONCLUSIONS: Carbohydrate in the ileum regulates postprandial exocrine pancreatic enzyme secretion and other postprandial upper gut functions. Peptide YY may play a role in this regulation. PMID- 7498668 TI - Intra-abdominal mass associated with gastrointestinal hemorrhage: a new manifestation of bacillary angiomatosis. AB - Bacillary angiomatosis is a recently described vascular proliferative lesion that occurs most commonly in individuals infected with human immunodeficiency virus. Cutaneous lesions are the most frequently described manifestations of bacillary angiomatosis. However, as culture techniques and disease recognition have improved, additional manifestations have been identified in human immunodeficiency virus-infected individuals, including bacillary peliosis hepatis and isolated bacteremia. Two species of the genus Bartonella (formerly Rochalimaea), Bartonella henselae or Bartonella quintana, have been cultured from the cutaneous lesions of bacillary angiomatosis. A new manifestation of Bartonella infection is reported: an intra-abdominal mass presenting with massive gastrointestinal hemorrhage in a patient with human immunodeficiency virus infection. B. quintana was cultured from a percutaneous needle-biopsy specimen obtained from the highly vascularized intra-abdominal mass. The bacillary angiomatosis lesion resolved after 3 months of tetracycline treatment. Recognition of Bartonella infection is extremely important because it is readily treatable with antibiotic therapy. PMID- 7498669 TI - Diagnosis of Wilson's disease in an asymptomatic sibling by DNA linkage analysis. AB - The molecular genetic diagnosis of Wilson's disease in the 5-year-old sister of a patient with Wilson's disease is reported. The girl was clinically free of disease and had no conventional biochemical markers of Wilson's disease (i.e., normal ceruloplasmin, normal copper in the serum, normal 24-hour urinary copper excretion). Diagnosis with restriction fragment length polymorphisms and a nonradioactive polymerase chain reaction-based analysis with microsatellite markers showed her to be homozygous for the disease-associated markers. A liver biopsy was performed, and a 20-fold increased liver copper content confirmed the diagnosis. The child was treated with chelation therapy with D-penicillamine. The report of this study clearly shows the advantage of DNA linkage analysis (especially polymerase chain reaction) over conventional laboratory methods for presymptomatic diagnosis of Wilson's disease before irreparable liver and neurological damage occurs. The only limitation of this DNA-based diagnosis is the fact that it is only applicable in siblings of an index patient whose diagnosis was made by phenotypic criteria. PMID- 7498670 TI - Reactive oxygen and nitrogen metabolites as mediators of secretory diarrhea. PMID- 7498671 TI - Irritable bowel syndrome: a multibillion-dollar problem. PMID- 7498672 TI - Microsatellite instability as an indicator of hereditary susceptibility to colon cancer. PMID- 7498673 TI - Malignant colorectal polyps. PMID- 7498674 TI - Primary chemoprevention strategies for colorectal cancer: ursodeoxycholic acid and other agents. PMID- 7498675 TI - Wherefore art thou guanylin? PMID- 7498676 TI - The selling of acetaminophen. PMID- 7498677 TI - Crohn's disease: to lump or to split? PMID- 7498678 TI - Antineutrophil cytoplasmic antibodies in autoimmune hepatitis. PMID- 7498679 TI - Bacteria and pathogenesis of gallstones. PMID- 7498680 TI - Serological testing for celiac disease in the elderly. PMID- 7498681 TI - Perspectives on esophageal manometry: "lumpers versus splitters". PMID- 7498682 TI - Biliary endoprostheses in elderly patients with endoscopically irretrievable common bile duct stones: report on 117 patients. AB - BACKGROUND: Endoscopic sphincterotomy and stone extraction can clear the bile ducts in 85% to 90% of patients. In case endoscopic stone extraction fails, insertion of a biliary endoprosthesis is an alternative. METHODS: Biliary endoprostheses were inserted in 117 patients (73 women and 44 men, median age 80 years). Follow-up was achieved by contacting referring physicians and general practitioners. RESULTS: Biliary drainage was established in all patients. Early complications occurred in 10 patients (9%) with a fatal outcome in 1. In 59 patients the endoprosthesis was a temporary measure before elective surgery or repeat endoscopy. Further treatment in these 59 patients caused only mild complications and there were no deaths. Endoprostheses were inserted as permanent therapy in 58 patients. Median follow-up in these 58 patients was 36 months (range, 1 to 117 months). A total of 34 complications occurred in 23 patients (40%), cholangitis being the most frequent. During follow-up, 44 patients died, 9 as a result of a biliary-related cause. CONCLUSIONS: These data favor temporary use of biliary endoprostheses in patients with endoscopically irretrievable bile duct stones. However, as a permanent therapy, late complications occur in many patients and the risk increases proportionally in time. Therefore, permanent biliary stenting should preferably be restricted to patients unfit for elective treatment at a later stage and with a short life expectancy. PMID- 7498684 TI - Endoscopic transpapillary drainage of pancreatic pseudocysts. AB - BACKGROUND: Endoscopic therapy of pancreatic pseudocysts has been reported mainly in small series. METHODS: The results of endoscopic transpapillary cyst drainage (ETCD) were evaluated prospectively in 30 patients with pancreatic pseudocysts. RESULTS: There were 24 men and 6 women with an average age of 45 years (SD 16). Twenty-eight had chronic pancreatitis (25 with alcoholic pancreatitis). Transpapillary cystopancreatic stents, with the tip into the cyst cavity, were inserted in 12 patients. Pancreatic stents with the tip as close as possible from the cyst cavity were inserted in the remaining 18 patients. Ten patients underwent an additional endoscopic cystenterostomy. The average duration of stenting was 4.4 months (range 15 days to 12 months). Patients were followed up for 15 months (range 2 to 60 months). All pseudocysts communicated with the pancreatic ductal system. The size of the pseudocysts ranged from 15 to 120 mm (average 50 mm). Pseudocysts were mainly located in the head of the pancreas (17 cases). Four minor complications occurred. There were no deaths. Twenty-six patients had pseudocyst resolution by ETCD, but 7 ultimately required surgery, 3 for early recurrence and 4 for failure of initial therapy. CONCLUSION: ETCD appears to be a safe and efficient modality for the drainage of pancreatic pseudocysts communicating with the pancreatic ductal system. PMID- 7498683 TI - The efficacy of endoscopic treatment of pancreatic pseudocysts. AB - BACKGROUND: Endoscopic drainage of pancreatic pseudocysts is a new nonsurgical treatment modality. We retrospectively studied the efficacy of endoscopic drainage of pseudocysts in 37 patients with chronic pancreatitis. METHODS: Endoscopic retrograde pancreatic drainage was performed in 12 patients, endoscopic cystogastrostomy in 10 patients, and endoscopic cystoduodenostomy in 7 patients. In the remaining 8 patients, combinations of drainage routes were used. RESULTS: ECG failed in 3 patients. Procedure-related complications were seen in 6 patients: bleeding in 3, perforation in 2, and apnea in 1 patient. There was no procedure-related mortality. Seven patients had complications in relation to stents or drains: pseudocyst infection due to stent clogging in 2, stent migration in 4, and kinking of the drain in 1 patient. Twenty-four patients had complete resolution of pseudocysts, 7 had partial resolution, and 6 had no resolution. Three patients had pseudocyst recurrences. Mean follow-up was 32 months. Finally, 10 patients underwent surgery. CONCLUSIONS: Endoscopic drainage was technically feasible in 92% of the patients. Procedure-related morbidity was 16% and mortality was 0%. Endoscopic drainage was a definitive treatment for two thirds of the patients (65%). Surgery can be reserved for those patients in whom endoscopic therapy fails. PMID- 7498685 TI - Treatment of pancreatic pseudocysts with ductal communication by transpapillary pancreatic duct endoprosthesis. AB - BACKGROUND: Endoscopic treatment of pancreatic pseudocysts via cystenterostomy has been recognized as a successful treatment option in carefully selected patients. Pancreatic transpapillary stenting as an alternative treatment option in patients with pancreatic pseudocysts directly communicating with the main duct has received little consideration. The aim of the current study was to assess the safety and utility of transpapillary pancreatic endoprosthesis in the treatment of communicating pseudocysts. METHODS: Twenty-one patients underwent placement of 33 transpapillary endoprostheses for the treatment of symptomatic pancreatic pseudocysts. All pseudocysts communicated with the main pancreatic duct and ranged in size from 3 to 9 cm (mean 6 cm). Eight patients had associated pancreatic duct strictures. RESULTS: Stent placement was successful in all cases: 13 directly into the pseudocyst, 8 beyond the stricture but not into the pseudocyst. Initial resolution of pseudocysts was seen in 17 patients, with 16 patients free of pseudocyst recurrence at mean follow-up of 37 months. All patients with associated strictures were treated successfully. Factors predictive of success included presence of strictures, size of pseudocyst greater than or equal to 6 cm, location in the body of the pancreas, and duration of pseudocyst less than 6 months. Complications included one episode of mild pancreatitis. CONCLUSIONS: Endoscopic treatment of symptomatic pancreatic pseudocysts with ductal communication by transpapillary pancreatic duct stenting is a safe, effective modality and should be considered a first line therapy. PMID- 7498686 TI - Transpapillary and transmural drainage of pancreatic pseudocysts. AB - BACKGROUND: Endoscopic drainage of pseudocysts using the transpapillary and transmural approaches has been reported. We evaluated endoscopic drainage in 53 patients with symptomatic pancreatic pseudocysts in whom conservative management had failed. METHODS: After preliminary endoscopic retrograde pancreatography, transpapillary drainage was attempted in 33 patients with pseudocysts that communicated with the main pancreatic duct. Transmural drainage of pseudocysts in contact with the stomach or duodenal wall was attempted in the remaining 20 patients and in 4 patients selected for combined transpapillary and transmural drainage. The cause of pseudocysts was chronic pancreatitis in 92%. The median pseudocyst size was 7.0 cm (range, 2 to 16). RESULTS: Endoscopic drainage was technically successful in 50 patients (94%), of whom 47 had complete pseudocyst resolution. Complications occurred in 11% and included gallbladder puncture (n = 1) and bleeding (n = 2) after transmural drainage, and pancreatitis (n = 1) after transpapillary drainage; stent clogging resulted in abscess formation in 2 patients. Mean follow-up was 22 months (range, 1 to 70); pseudocysts recurred in 11 patients (23%), of whom 7 were successfully re-treated endoscopically. CONCLUSION: Both transpapillary and transmural pseudocyst drainage are highly effective in patients with pseudocysts demonstrating suitable anatomy for these endoscopic techniques. PMID- 7498687 TI - Value of endoscopic ultrasonography in the diagnosis of common bile duct stones: comparison with surgical exploration and ERCP. AB - An accurate and safe preoperative method of imaging the common bile duct is essential for the proper diagnosis of calculous biliary tract disease, especially in the current era of laparoscopic cholecystectomy. The value of endoscopic ultrasonography in detecting common duct stones has been reported, albeit in small series. The aim of this retrospective study was to assess the accuracy of EUS in a large series of patients. We compared EUS to direct cholangiography in the evaluation of 422 patients for common duct stones. Ductal stones were imaged by EUS in 168 patients (43.4%). No complications were encountered. EUS failed in 2.3% of cases, ERCP failed in 8.3%, and surgical exploration failed in 0.5%. Comparison of EUS with surgical exploration in 185 patients showed a sensitivity of 94.9%, a specificity of 97.8%, and an accuracy of 95.9%. EUS was compared to ERCP in 219 patients. All common duct stones found by ERCP were evident by EUS. Concordance was obtained in 91.3% of cases. Review of videotapes disclosed 3 false-positives and 16 unequivocal true-positives. We conclude that EUS is a safe and highly accurate means of detecting common duct stones and should be proposed before laparoscopic cholecystectomy in patients at risk of choledocholithiasis. PMID- 7498688 TI - Patchiness of mucosal inflammation in treated ulcerative colitis: a prospective study. AB - Conventional wisdom dictates that ulcerative colitis affects contiguous areas of the colon and is most severe in the rectum, and that the finding of rectal sparing or patchy involvement should raise suspicions of Crohn's disease. We and others have noted occasional rectal sparing and patchy involvement in patients with ulcerative colitis. Therefore, we prospectively studied the prevalence of patchiness, including rectal sparing, in treated cases of ulcerative colitis. Consecutive patients with longstanding ulcerative colitis were studied. The left colon was divided into three zones for scoring degree of activity, and biopsy specimens from each zone were graded for histologic activity by a blinded observer. Patchiness by endoscopy or histology was defined as (1) frank rectal sparing (normal appearance endoscopically; absence of inflammation of the lamina propria and crypts histologically); (2) areas of greater inflammation proximally than distally; or (3) discrete areas of patchiness endoscopically within any one zone. Of 39 patients evaluated, 17 (44%) had endoscopic evidence of patchiness, including 5 (13%) with rectal sparing. Thirteen (33%) had histologic evidence of patchiness, including 6 (15%) with rectal sparing. Both endoscopic and histologic patchiness were seen in 9 patients (23%). The patchy and nonpatchy groups did not differ in regard to the use of rectal therapy. In patients with treated ulcerative colitis, the finding of rectal sparing or patchiness should not necessarily indicate a change in the diagnosis to Crohn's disease. PMID- 7498689 TI - Single-day, divided-dose oral sodium phosphate laxative versus intestinal lavage as preparation for colonoscopy: efficacy and patient tolerance. AB - Polyethylene glycol-electrolyte lavage solutions are widely used to prepare the colon for colonoscopy. Unfortunately, some patients find this preparation difficult to complete. Recent studies of a sodium phosphate-based laxative have shown both good patient tolerance and good bowel preparation. In these studies, the laxative has generally been prescribed in two doses, with the second dose taken early the morning of colonoscopy. Because the morning dose is inconvenient for many patients, we compared giving a common polyethylene glycol-based electrolyte lavage solution the day before colonoscopy with our method of giving both doses of sodium phosphate-based laxative the day before colonoscopy: one dose at 4 PM and the second dose at 8 PM. We judged efficacy by an assessment of residual liquid and fecal matter in the colon and judged tolerance by the results of a symptom questionnaire completed by each patient immediately before the procedure. Our results in more than 200 patients showed similar efficacy ratings and similar symptom scores for both preparations, but patients rated the sodium phosphate-based preparation as easier to tolerate. In conclusion, in selected patients this new dosing method for sodium phosphate is preferable to large volume, whole-gut lavage solutions. PMID- 7498690 TI - Endoscopic reduction of gastric volvulus: the alpha-loop maneuver. PMID- 7498691 TI - Endosonography probe-guided endoscopic mucosal resection of gastric neoplasms. PMID- 7498692 TI - Endoscopic treatment of stenosis in recurrent Crohn's disease with balloon dilation combined with local corticosteroid injection. PMID- 7498695 TI - An unusual presentation of pill-induced esophagitis. PMID- 7498694 TI - Decrease in a markedly elevated CA19-9 level after stenting of a benign pancreatic ductal stricture. PMID- 7498693 TI - Miniature ultrasonic probe evaluation of esophageal varices after endoscopic variceal ligation. PMID- 7498696 TI - Coexistence of anomalous pancreaticobiliary ductal union with adenomyomatosis of the gallbladder. PMID- 7498697 TI - Distal small bowel obstruction by a severed PEG tube: successful endoscopic management by ileoscopic retrieval. PMID- 7498698 TI - Stents for stones: short-term good, long-term uncertain. PMID- 7498699 TI - Endoscopic management of pancreatic pseudocysts continues to evolve. PMID- 7498700 TI - Supplemental oxygen during endoscopy in patients with ischemic heart disease. PMID- 7498701 TI - Antibiotic therapy of cholangitis. PMID- 7498702 TI - Bipolar electrocautery as an alternative to Nd:YAG laser for the treatment of the watermelon stomach. PMID- 7498703 TI - Safe maneuvering of echoendoscopes in patients with distorted duodenal anatomy. PMID- 7498704 TI - Repositioning of a migrated esophageal stent using a retroflexed endoscope. PMID- 7498705 TI - Retrograde migration of PEG tubes. PMID- 7498706 TI - External PEG button migration. PMID- 7498707 TI - Randomized trial of endoscopic stenting versus surgical bypass in malignant low bile duct obstruction. PMID- 7498709 TI - Pre-endoscopy screening using serodiagnosis of Helicobacter pylori infection. PMID- 7498708 TI - Serum lidocaine and MEGX concentrations after pharyngeal anesthesia for gastroscopy. PMID- 7498710 TI - [Between tradition and progress--Turkish fathers as labor participants]. PMID- 7498711 TI - [Help or hinderniss? The effect of legal norms on the physician-patient relationship]. PMID- 7498712 TI - [Medical management after spontaneous abortion]. PMID- 7498713 TI - [The fate of women reflected in classical literature]. PMID- 7498714 TI - [Determination of cytokines and cytokine receptors in premature labor]. AB - One of the leading causes of preterm labour is a subclinical intrauterine infection. The diagnosis of this inapparent infection is an unsolved problem, because clinical parameters show the infection mostly too late and cannot be used for early detection of women at high risk for preterm labour. A prospective clinical study measuring cytokines and cytokine receptor concentration in amniotic fluid from patients with preterm labour and preterm rupture of membranes (PROM) was undertaken to answer the question whether the amount of cytokines is positively correlated to the risk of preterm delivery. In 78 patients (43 controls, 23 patients with PROM, 7 with preterm labour and delivery < 37 weeks of gestation, 7 with preterm labour and delivery at term) amniotic fluid was collected and the following cytokines were measured by an ELISA: IL1 beta, IL2, IL6, TNF alpha, IFN gamma, TNF receptor 55 und 75, IFN gamma receptor, IL2 receptor. We found significant, in some cases highly significant differences in the cytokine content of amniotic fluid of patients with PROM compared with controls and patients with preterm labour and delivery before 37 weeks of gestation compared with controls. There were no differences between patients with preterm labour and delivery at term and controls. Amniotic fluid cultures were positive only in 20-25% of the cases and therefore not a predictive sign. The determination of cytokines in amniotic fluid of patients with preterm labour seems to be a good marker to predict the risk of preterm delivery.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7498715 TI - [Value of serum placental protein 14 determination in differential diagnosis of abnormal early pregnancy]. AB - Single measurements of hCG and placental protein 14 were carried out in 32 patients with tubal pregnancies, in 38 patients with spontaneous abortion and 28 women with normal intrauterine pregnancies from the 5th to the 7th week of pregnancy. HCG-measurement could only show significant differences between normal and abnormal early pregnancies. PP 14-measurements showed significant differences between all groups. Low concentrations of serum PP 14 point to an ectopic pregnancy. PMID- 7498716 TI - [Idiopathic habitual abortion: experiences with active immunotherapy]. AB - The effect of lymphocyte transfusions on the stabilisation and successful outcome of pregnancy was investigated in 30 women with recurrent abortions presumably caused by immunologic factors. Lymphocyte concentrates were obtained from stored whole blood of the husbands and transferred by intravenous injection. Exclusion criteria for this study were the presence of antipaternal lymphocytotoxic antibodies in the patient's serum (cross match) or an advanced pregnancy (> 12 weeks). After transfusion of paternal lymphocytes 19 (63%) women had uncomplicated and successful pregnancies whereas 11 (37%) suffered again from early pregnancy loss. Two out of these 11 patients aborted twice. The probability of a successful treatment decreased as soon as lymphocytotoxic antibodies appeared in the patient's serum (p < or = 0.021). Women with a lower number of abortions before treatment carried their pregnancies to full term more frequently (p < or = 0.03). The number of transfusions, the period between start of treatment and conception as well as the distribution of age within the two groups had no significant effect on the outcome of therapy. A lack of lymphocytotoxic antibodies or their late appearance during treatment give a positive prognosis for the progress of pregnancy. Compared with a fertile control group, couples with habitual abortions showed no significantly increased HLA sharing. Immunisation with paternal lymphocytes in cases of presumably habitual abortions due to immunologic factors appears as an effective therapy with only few side effects. Large, randomised, double-blind, and placebo-controlled trials are needed before drawing conclusions. PMID- 7498717 TI - [Effect of pregnancy and labor on serotonin metabolism]. AB - Several complications of pregnancy have been reported to be associated with alterations of serotonin (5-hydroxytryptamine; 5-HT) metabolism. Reported data regarding changes of 5-HT metabolism in pregnancy are partially contradictory. The presented study tries to describe pregnancy-associated changes in 5-HT metabolism. Urine and blood samples were taken from 20 pregnant women 4 weeks before, within 24 hours and on the 4th day following delivery. 17 non-pregnant age-matched women served as controls. 5-HT and its main metabolite 5 hydroxyindoleacetic acid (5-HIAA) were analysed. We observed a continuously increasing production and excretion of unbound 5-HT and 5-HIAA during pregnancy. Four days after delivery 5-HT production returned to baseline levels, while excretion remained elevated. 5-HIAA excretion increased in pregnancy and return to baseline levels on the day following delivery. Increased 5-HT metabolism and excretion results in a decreased 5-HT serum concentration despite increased 5-HT production during pregnancy. Our results help to understand the contradictory observations of elevated 5-HT production on the one hand and decreased 5-HT activity during pregnancy on the other. PMID- 7498718 TI - [Legal problems in obstetrics. On the "liability of the expert witness"]. AB - There has been a depressing increase in claims for damages and associated civil code and legal code proceedings in medicine in general (as the term "defensive medicine" illustrates) that exercises considerable influence on medical activities in obstetrics, not always to the benefit of the patients. In what manner and to what extent the expert can and must influence legal decisions, is demonstrated by means of a few examples such as management of parturition in case of breech presentation, causal assessment of psychomotoric retardation in postnatal life and the management of dystocia in shoulder presentation. If judges and attorneys are blamed-as is often the case-one should always also consider the significance of expertising activities. Typical errors occurring during expert assessment are pointed out. This shows the high degree of responsibility that has to be shouldered by the expert when he advises judges and/or attorneys. Expertising activities should be directed to a greater measure than in recent years, at delivering expertises restricted to the factual situation without involving emotions. This would at the same time reduce the influence of judges, attorneys and lawyers on our medical activities, an influence that has recently been much deplored. PMID- 7498719 TI - [Chlamydia infections in gynecology and obstetrics]. AB - Urogenital infections with Chlamydia trachomatis belong to the most prevalent sexual transmitted diseases worldwide. In women, Chlamydia trachomatis can cause an infection of the cervix, the urethra, the endometrium and the Fallopian tubes. As potential sequelae, tubal infertility and ectopic pregnancies are associated with urogenital infections caused by this pathogen. An infection of the cervix in pregnancy can lead to neonatal and maternal complications. Therefore, by April 1st, 1995 a cervical swab for Chlamydia trachomatis has been included in the German antenatal care guide lines. The significance of these infections in health care is in contrast to its often hardly symptomatic or asymptomatic course. New diagnostic methods facilitate the detection of the pathogen and allow a specific antibiotic therapy. The aim of this review is a presentation of the clinical significance, diagnosis and therapy of urogenital infections with Chlamydia trachomatis in obstetrics and gynaecology to contribute to the prevention of potentially severe sequelae. PMID- 7498720 TI - [Cytogenetic abnormalities in sperm: a risk factor for application of medical reproduction techniques?]. AB - Microinsemination techniques are increasingly applied to achieve fertilisation of the oocyte in cases of male fertility disorders. Thus, spermatozoa can directly interact with the oolemma or the ooplasm, thereby obviating the need for penetration through cumulus oophorus and zona pellucida. However, problems associated with this particular feature arise in view of the following considerations: 1. The gametes from subfertile males are suspected to carry more cytogenetic anomalies in addition to an impaired sperm quality when compared to fertile men. 2. Cumulus and zona play an important role in the selection of morphologically abnormal and dysfunctional spermatozoa. 3. Consequently, bypassing natural sperm selection processes would lead to more abnormal embryos, abortions, and births of diseased or handicapped children. Spermatozoa can be analysed cytogenetically after fusion with zona-free hamster eggs or by fluorescence in situ hybridization (FISH). Both techniques did not confirm increased aberrations in gametes from subfertile men with a normal somatic karyotype. However, a definite conclusion cannot be drawn because of the limited data and restrictions concerning their interpretation. This also holds true for the evaluation of correlations indicating no significant relationship between the chromosomal constitution and sperm motility or morphology, respectively. Currently, the risk of malformation after microinsemination does not appear to be increased when compared to conventional IVF but the results do not yet allow a reliable statistical evaluation. In contrast, decreased morphological quality, impaired implantation, and early abortions have been reported for embryos produced by assisted fertilisation. This suggests that an effective natural selection of abnormal embryos will prevent a significant increase in malformations in the following generation.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7498721 TI - [Intracytoplasmic versus subzonal spermatozoa injection. A comparison of 2 different methods of micro-assisted fertilization]. AB - In order to define the usefulness of subzonal sperm insertion (SUZI) and intracytoplasmic sperm injection (ICSI), we studied in a comparative trial 46 consecutive treatment cycles of microassisted fertilisation by SUZI and ICSI. By ICSI 9% of the oocytes in metaphase II were damaged in 26 treatment cycles, by SUZI, however, not one single egg in 20 cycles (p < 0.001). Fertilisation rate after ICSI (65%) was, particularly in cases with combined sperm defects as well, significantly higher than after SUZI (35%; p < 0.001). Additionally, after ICSI a higher transfer (100% vs. 75%; p < 0.05) and pregnancy rate could be obtained (38% vs. 10% per cycle; p < or = 0.05). At the moment, 3 healthy children are born (2 after SUZI, 1 after ICSI), 4 patients are in the 2nd and 3rd trimenon, respectively, the remaining 5 patients aborted. In conclusion, the ICSI technique yields better results than SUZI, especially in cases of very severe male subfertility. PMID- 7498722 TI - [Smoking--a possible cause for failure in IVF/ET treatment?]. AB - The aim of this study was to investigate the relation between cigarette smoking of women and the fertilisation and pregnancy rates obtained by in-vitro fertilisation treatment. To guarantee an objective recording of tobacco smoke exposure, the smoking habit was not determined by questionnaires, but by cotinine, the principal metabolite of nicotine. The study population consisted of 197 women (23-39 years old) who were divided into the groups 'non-smokers' (n = 68), 'passive smokers' (n = 26) and 'active smokers' (n = 103) according to the cotinine concentration measured in follicular fluid. The reason for infertility was strictly a tubal factor with apparently normal ovulatory cycles. There were no significant differences in fertilisation and pregnancy rates between the different groups (p > 0.05). The E2 serum levels were significantly decreased in women who smoked, compared with the results obtained from non-smokers and passive smokers (p < 0.025). Overall, a strong negative correlation of the cotinine- and E2 levels was observed (r = -0.65). The results suggest that there is no clinically detectable impairment of fertilisation potential due to female smoking and that there is a greater influence on the outcome of in-vitro fertilisation by other factors. PMID- 7498723 TI - [Triplet pregnancy after intracytoplasmic sperm injections]. AB - Morphology of the spermatozoa is one of the major criteria in severe male subfertility. We report on a case of severe Oligoasthenoteratozoospermia in which a triplet pregnancy could be achieved after intracytoplasmic sperm injection (ICSI). After treatment with GnRH-analogues/HMG/HCG seven oocytes of metaphase II could be retrieved and injected. Three of them fertilised and were transferred as four-cell stage embryos after two days. Seven weeks later an intact triplet pregnancy could be detected by ultrasonography. The severe sperm defects in this case were evaluated with the technique of electron microscopy. Ultramorphology of spermatozoa appears to have an important predictive value for fertilisation and pregnancy in classical IVF. Its value for the assisted procreation techniques such as ICSI remains to be investigated. PMID- 7498724 TI - Quarreling geneticists and a diplomat. PMID- 7498725 TI - Role of Erf recombinase in P22-mediated plasmid transduction. AB - In the absence of host RecA function, plasmid transduction by bacteriophage P22 can be mediated by Erf recombinase. Erf is not carried on the infecting particle but synthesized upon infection. In the recipient cell, Erf can promote both generalized plasmid transduction (which requires the circularization of plasmids transduced as linear multimers) and specialized plasmid transduction (which requires the release of plasmid DNA from linear plasmid-phage cointegrates). Both processes of Erf-mediated plasmid transduction require host RecBCD function. In contrast, RecBCD is not required for Erf-mediated circularization of P22 DNA. PMID- 7498726 TI - Mutations in the mitochondrial ATP synthase gamma subunit suppress a slow-growth phenotype of yme1 yeast lacking mitochondrial DNA. AB - In Saccharomyces cerevisiae, inactivation of the nuclear gene YME1 causes several phenotypes associated with impairment of mitochondrial function. In addition to deficiencies in mitochondrial compartment integrity and respiratory growth, yme1 mutants grow extremely slowly in the absence of mitochondrial DNA. We have identified two genetic loci that, when mutated, act as dominant suppressors of the slow-growth phenotype of yme1 strains lacking mitochondrial DNA. These mutations only suppressed the slow-growth phenotype of yme1 strains lacking mitochondrial DNA and had no effect on other phenotypes associated with yme1 mutations. One allele of one linkage group had a collateral respiratory deficient phenotype that allowed the isolation of the wild-type gene. This suppressing mutation was in ATP3, a gene that encodes the gamma subunit of the mitochondrial ATP synthase. Recovery of two of the suppressing ATP3 alleles and subsequent sequence analysis placed the suppressing mutations at strictly conserved residues near the C terminus of Atp3p. Deletion of the ATP3 genomic locus resulted in an inability to utilize nonfermentable carbon sources. atp3 deletion strains lacking mitochondrial DNA grew slowly on glucose media but were not as compromised for growth as yme1 yeast lacking mitochondrial DNA. PMID- 7498727 TI - Specificities of the Saccharomyces cerevisiae rad6, rad18, and rad52 mutators exhibit different degrees of dependence on the REV3 gene product, a putative nonessential DNA polymerase. AB - The Saccharomyces cerevisiae rad6, rad18, and rad52 mutants exhibit DNA repair deficiencies and distinct mutator phenotypes. DNA replication past unrepaired spontaneous damage might contribute to the specificities of these mutators. Because REV3 is thought to encode a DNA polymerase that specializes in translesion synthesis, we determined the REV3 dependence of the rad mutator specificities. Spontaneous mutagenesis at a plasmid-borne SUP4-o locus was examined in isogenic strains having combinations of normal or mutant REV3 and RAD6, RAD18, or RAD52 alleles. For the rad6 and rad18 mutators, the mutation rate increase relied largely, but not exclusively, on REV3 whereas the rad52 mutator was entirely REV3 dependent. The influence of REV3 on the specificity of the rad6 mutator differed markedly depending on the mutational class examined. However, the requirement of rev3 for the production of G.C-->T.A transversions by the rad18 mutator, which induces only these substitutions, was similar to that for rad6-mediated G.C-->T.A transversion. This supports a role for the Rad6-Rad18 protein complex in the control of spontaneous mutagenesis. The available data imply that the putative Rev3 polymerase can process a variety of spontaneous DNA lesions that normally are substrates for error-free repair. PMID- 7498729 TI - Hot spots of recombination in fission yeast: inactivation of the M26 hot spot by deletion of the ade6 promoter and the novel hotspot ura4-aim. AB - The M26 mutation in the ade6 gene of Schizosaccharomyces pombe creates a hot spot of meiotic recombination. A single base substitution, the M26 mutation is situated within the open reading frame, near the 5' end. It has previously been shown that the heptanucleotide sequence 5' ATGACGT 3', which includes the M26 mutation, is required for hot spot activity. The 510-bp ade6-delXB deletion encompasses the promoter and the first 23 bp of the open reading frame, ending 112 bp upstream of M26. Deletion of the promoter in cis to M26 abolishes hot spot activity, while deletion in trans to M26 has no effect. Homozygous deletion of the promoter also eliminates M26 hot spot activity, indicating that the heterology created through deletion of the promoter per se is not responsible for the loss of hot spot activity. Thus, DNA sequences other than the heptanucleotide 5' ATGACGT 3', which must be located at the 5' end of the ade6 gene, appear to be required for hot spot activity. While the M26 hotspot stimulates crossovers associated with M26 conversion, it does not affect the crossover frequency in the intervals adjacent to ade6. The flanking marker ura4-aim, a heterology created by insertion of the ura4+ gene upstream of ade6, turned out to be a hot spot itself. It shows disparity of conversion with preferential loss of the insertion. The frequency of conversion at ura4-aim is reduced when the M26 hot spot is active 15 kb away, indicating competition for recombination factors by hot spots in close proximity. PMID- 7498728 TI - sck1, a high copy number suppressor of defects in the cAMP-dependent protein kinase pathway in fission yeast, encodes a protein homologous to the Saccharomyces cerevisiae SCH9 kinase. AB - Schizosaccharomyces pombe regulates intracellular cAMP levels, and thus cAMP dependent protein kinase (PKA) activity, in response to changes in nutrient conditions. Mutations in any of eight git genes inhibit glucose repression of fbp1 transcription, alter the cell morphology, and cause a reduction in the growth rate. The eight git genes encode components of an adenylate cyclase activation pathway, adenylate cyclase itself, and the catalytic subunit of PKA. Three of these genes have been identified in other studies as regulators of meiosis. Here we show that the sck1 gene, cloned as a high copy number suppressor of a mutation in git3, is able to suppress the defects conferred by a mutation in any of these git genes. Sequence analysis suggests that sck1 encodes a protein most closely related to the Saccharomyces cerevisiae SCH9 protein kinase that had previously been identified as a high copy number suppressor of mutations in S. cerevisiae that reduce or eliminate PKA activity. Disruption of the sck1 gene causes a significant delay in exit from stationary phase when combined with a disruption of the pka1 (git6) gene encoding the catalytic subunit of PKA. However, the sck1 disruption by itself has little or no effect upon fbp1 transcription, meiosis, or exit from stationary phase, and does not enhance the constitutive fbp1 transcription observed in a pka1 mutant. Therefore, sck1 appears to function in a redundant fashion to pka1, but to varying degrees, in the pathways regulated by pka1. PMID- 7498730 TI - Genetic and physical mapping of telomeres in the rice blast fungus, Magnaporthe grisea. AB - Telomeric restriction fragments were genetically mapped to a previously described linkage map of Magnaporthe grisea, using RFLPs identified by a synthetic probe. (TTAGGG)3. Frequent rearrangement of telomeric sequences was observed in progeny isolates creating a potential for misinterpretation of data. Therefore a consensus segregation data set used to minimize mapping errors. TWelve of the 14 telomeres were found to be genetically linked to existing RFLP markers. Second dimensional electrophoresis of restricted chromosomes confirmed these linkage assignments and revealed the chromosomal location of the two unlinked telomeres. We were thus able to assign all 14 M. grisea telomeres to their respective chromosome ends. The Achilles' cleavage (AC) technique was employed to determine that chromosome 1 markers 11 and CH5-120H were approximately 1.8 Mb and 1.28 Mb, respectively, from their nearest telomeres. RecA-AC was also used to determine that unlinked telomere 6 was approximately 530 kb from marker CH5-176H in strain 2539 and 580 kb in Guy11. These experiments indicated that large portions of some chromosome ends are unrepresented by genetic markers and provided estimates of the relationship of genetic to physical distance in these regions of the genome. PMID- 7498731 TI - Altered mating-type identity in the fungus Podospora anserina leads to selfish nuclei, uniparental progeny, and haploid meiosis. AB - In wild-type crosses of the filamentous ascomycete Podospora anserina, after fertilization, only nuclei of opposite mating type can form dikaryons that undergo karyogamy and meiosis, producing biparental progeny. To determine the role played by the mating type in these steps, the four mat genes were mutagenized in vitro and introduced into a strain deleted for its mat locus. Genetic and cytological analyses of these mutant strain, crossed to each other and to wild type, showed that mating-type information is required for recognition of nuclear identity during the early steps of sexual reproduction. In crosses with strain carrying a mating-type mutation, two unusual developmental patterns were observed: monokaryotic cells, resulting in haploid meiosis, and uniparental dikaryotic cells providing, after karyogamy and meiosis, a uniparental progeny. Altered mating-type identity leads to selfish behavior of the mutant nucleus: it migrates alone or paired, ignoring its wild-type partner in all mutant x wild type crosses. This behavior is nucleus-autonomous because, in the same cytoplasm, the wild-type nuclei form only biparental dikaryons. In P. anserina, mat genes are thus required to ensure a biparental dikaryotic state but appear dispensable for later stages, such as meiosis and sporulation. PMID- 7498732 TI - A novel positive selection for identifying cold-sensitive myosin II mutants in Dictyostelium. AB - We developed a positive selection for myosin heavy chain mutants in Dictyostelium. This selection is based on the fact that brief exposure to azide causes wild-type cells to release from the substrate, whereas myosin null cells remain adherent. This procedure assays myosin function on a time scale of minutes and has therefore allowed us to select rapid-onset cold-sensitive mutants after random chemical mutagenesis of Dictyostelium cells. We developed a rapid technique for determining which mutations lie in sequences of the myosin gene that encode the head (motor) domain and localized 27 of 34 mutants to this domain. We recovered the appropriate sequences from five of the mutants and demonstrated that they retain their cold-sensitive properties when expressed from extrachromosomal plasmids. PMID- 7498733 TI - The ornithine decarboxylase gene of Caenorhabditis elegans: cloning, mapping and mutagenesis. AB - The gene (odc-1) encoding ornithine decarboxylase, a key enzyme in polyamine biosynthesis, was cloned and characterized. Two introns interrupt the coding sequence of the gene. The deduced protein contains 422 amino acids and is homologous to ornithine decarboxylases of other eukaryotic species. In vitro translation of a transcript of the cDNA yielded an enzymatically active product. The mRNA is 1.5 kb in size and is formed by trans-splicing to SL1, a common 5' RNA segment. odc-1 maps to the middle of LG V, between dpy-11 and unc-42 and near a breakpoint of the nDf32 deficiency strain. Enzymatic activity is low in starved stage 1 (L1) larva and, after feeding, rises progressively as the worms develop. Targeted gene disruption was used to create a null allele. Homozygous mutants are normally viable and show no apparent defects, with the exception of a somewhat reduced brood size. In vitro assays for ornithine decarboxylase activity, however, show no detectable enzymatic activity, suggesting that ornithine decarboxylase is dispensible for nematode growth in the laboratory. PMID- 7498734 TI - Caenorhabditis elegans mutants resistant to inhibitors of acetylcholinesterase. AB - We characterized 18 genes from Caenorhabditis elegans that, when mutated, confer recessive resistance to inhibitors of acetylcholinesterase. These include previously described genes as well as newly identified genes; they encode essential as well as nonessential functions. In the absence of acetylcholinesterase inhibitors, the different mutants display a wide range of behavioral deficits, from mild uncoordination to almost complete paralysis. Measurements of acetylcholine levels in these mutants suggest that some of the genes are involved in presynaptic functions. PMID- 7498735 TI - Competition between mitochondrial haplotypes in distinct nuclear genetic environments: Drosophila pseudoobscura vs. D. persimilis. AB - A test for coadaptation of nuclear and mitochondrial genomes was performed using the sibling species, Drosophila pseudoobscura and D. persimilis. Two lines of flies with "disrupted" cytonuclear genotypes were constructed by repeated backcrossing of males from one species to females carrying mitochondrial DNA (mtDNA) from the other species. Each "disrupted" strain was competed in population cages with the original stock of each species from which the recurrent males were obtained during the backcrossing. As such, the two species' mitochondrial types were competed reciprocally in the nuclear genetic environments of each species. The trajectories of mtDNA haplotypes were followed in discrete-generation population cages using a PCR-four-cutter approach. A significant increase in the frequency of D. pseudoobscura mtDNA was observed in each of four replicate cages with a D. pseudoobscura nuclear background. In the D. persimilis nuclear background, one cage actually showed an increase in frequency of D. pseudoobscura mtDNA, although together the four replicate cages show little change in frequency. These results were repeated after frequency perturbations and reinitiation of each cage. An analysis of fitness components revealed that fertility selection greatly outweighed viability selection in these cytonuclear competition experiments. The asymmetry of the fitnesses of the mtDNA haplotypes on the two genetic backgrounds is consistent in direction with the previously reported asymmetry of female fertility in backcrosses between these two species. While our experiments do not allow us to identify mtDNA as the sole source of fitness variation, at a minimum the data indicate a fitness association between nuclear fertility factors and the D. pseudoobscura mtDNA on its own genetic background. PMID- 7498736 TI - Temporal patterns of gene expression in the antenna of the adult Drosophila melanogaster. AB - The time course of gene expression in the adult fruit fly has been partially characterized by using enhancer trap and reporter gene constructs that mark 49 different genes. The relative intensity of the reporter protein in individual cells of the antennae was measured as a function of adult age. Most genes showed a graduated expression, and the intensity of expression had a reproducible and characteristic time course. Different genes displayed different temporal patterns of expression and more often than not the pattern of expression was complex. We found a number of genes having patterns that scaled with life span. In these cases the intensity of gene expression was found to be invariant with respect to biological time, when expressed as a fraction of the life span of the line. The scaling was observed even when life span was varied as much as threefold. Such scaling serves to (1) further demonstrate that deterministic mechanisms such as gene regulation act to generate the temporal patterns of expression seen during adult life, (2) indicate that control of these regulatory mechanisms is linked to life span, and (3) suggest mechanisms by which this control is accomplished. We have concluded that gene expression in the adult fly is often regulated in a fashion that allows for graduated expression over time, and that the regulation itself is changing throughout adult life according to some prescribed program or algorithm. PMID- 7498737 TI - Genetic analysis of embryonic cis-acting regulatory elements of the Drosophila homeotic gene sex combs reduced. AB - The homeotic gene Sex combs reduced (Scr) of Drosophila melanogaster is expressed in the labial and prothoracic segments of the ectoderm, in parasegments two and three of the CNS, and in the visceral mesoderm of the anterior and posterior midgut. The mutationally defined function of Scr is to specify the identity of the labial and prothoracic segments and to control the development of the gastric caeca. The Scr locus occupies a chromosomal region of approximately 80 kb within the Antennapedia complex (ANT-C). To understand how Scr's spatiotemporal expression pattern is generated in the embryo, we have mapped its transcriptional regulatory elements using three approaches. First, we examined the expression pattern of Scr in embryos containing chromosomal rearrangements that remove potential Scr regulatory elements. Second, we made and analyzed a set of Scr minigene transformants. Third, we analyzed a set of Scr-lacZ enhancer tester constructs. Using more sensitive anti-SCR antisera, we discovered that Scr is expressed in tissues that were not previously thought to accumulate SCR: a stripe of ectodermal cells in the parasegment 2 region of stage 5 embryos, the embryonic salivary glands, and the dorsal ridge. Four DNA fragments that had previously been shown in an analysis of Scr-lacZ reporter constructs to contain putative Scr enhancer elements were found to have functional enhancers; similarly, another Scr fragment was found to contain a functional repressor. Our results suggest that regulation of Scr in the labial segment and the CNS requires the apparently synergistic action of multiple, widely spaced enhancer elements. Regulation in the prothorax also appears to be controlled by multiple enhancers:one complete pattern element and one subpattern element. In contrast, Scr regulation in the visceral mesoderm is controlled by an enhancer(s) located in only one DNA fragment. PMID- 7498738 TI - Identification of homeotic target genes in Drosophila melanogaster including nervy, a proto-oncogene homologue. AB - In Drosophila, the specific morphological characteristics of each segment are determined by the homeotic genes that regulate the expression of downstream target genes. We used a subtractive hybridization procedure to isolate activated target genes of the homeotic gene Ultrabithorax (Ubx). In addition, we constructed a set of mutant genotypes that measures the regulatory contribution of individual homeotic genes to a complex target gene expression pattern. Using these mutants, we demonstrate that homeotic genes can regulate target gene expression at the start of gastrulation, suggesting a previously unknown role for the homeotic genes at this early stage. We also show that, in abdominal segments, the levels of expression for two target genes increase in response to high levels of Ubx, demonstrating that the normal down-regulation of Ubx in these segments is functional. Finally, the DNA sequence of cDNAs for one of these genes predicts a protein that is similar to a human proto-oncogene involved in acute myeloid leukemias. These results illustrate potentially general rules about the homeotic control of target gene expression and suggest that subtractive hybridization can be used to isolate interesting homeotic target genes. PMID- 7498741 TI - Developmental genetical analysis and molecular cloning of the abnormal oocyte gene of Drosophila melanogaster. AB - Studies of the abnormal oocyte (abo) gene of Drosophila melanogaster have previously been limited to the analysis of a single mutant allele, abnormal oocyte1 (abo1). The abo1 mutation causes a maternal-effect lethality that can be partially rescued zygotically by the abo+ allele and by increasing the dosage of specific regions of heterochromatin denoted ABO. This report describes the properties of abo2, a new P-element-induced allele that allowed us to reexamine the nature of maternal-effect defect. Comparisons of the phenotype of progeny of abo1/abo1 and abo1/abo2 females show that the preblastoderm lethality previously described as a component of the abo mutant maternal effect results from a recessive fertilization defect associated with the abo1 chromosome. We demonstrate here that the abo-induced maternal effect lethality occurs predominately late in embryogenesis after cuticle deposition but before hatching. The phenocritical period for zygotic rescue by heterochromatin coincides with this period of late embryogenesis. We have used the abo2 mutation to map and molecularly clone the gene. We show that the abo gene is located in the 32C cytogenetic interval and identify the putative abo transcript from mRNA isolated from adult females. Using germline transformation, we show that a 9-kb genomic fragment to which the transcript maps, partially fulfills requirement for maternal and zygotic abo+ function. PMID- 7498739 TI - The Suppressor of fused gene encodes a novel PEST protein involved in Drosophila segment polarity establishment. AB - Suppressor of fused, Su(fu), was identified as a semi-dominant suppressor of the putative serine/threonine kinase encoded by the segment polarity gene fused in Drosophila melanogaster. The amorphic Su(fu) mutation is viable, shows a maternal effect and displays no phenotype by itself. Su(fu) mutations are often found associated to karmoisin (kar) mutations but two complementation groups can be clearly identified. By using a differential hybridization screening method, we have cloned the Su(fu) region and identified chromosomal rearrangements associated with Su(fu) mutations. Two classes of cDNAs with similar developmental patterns, including a maternal contribution, are detectable in the region. Transformation experiments clearly assigned the Su(fu)+ function to one of these transcription units while the other one can be most likely assigned to the kar+ function. Surprisingly the 5' end of the kar RNA mapped within the 3' untranslated region of the Su(fu) transcribed sequence. The Su(fu) gene encodes a 53-kD protein, which contains a PEST sequence and shows no significant homologies with known proteins. Genetic analysis shows that proper development requires a fine tuning of the genetic doses of fu and Su(fu) both maternally and zygotically. These results, together with previous genetic and molecular data, suggest that fused and Suppressor of fused could act through a competitive posttraductionnal modification of a common target in the hedgehog signaling pathway. PMID- 7498740 TI - Functional studies of the carboxy-terminal repeat domain of Drosophila RNA polymerase II in vivo. AB - To understand the in vivo function of the unique and conserved carboxy-terminal repeat domain (CTD) of RNA polymerase II largest subunit (RpII215), we have studied RNA polymerase II biosynthesis, activity and genetic function in Drosophila RpII215 mutants that possessed all (C4), half (W81) or none (IIt) of the CTD repeats. We have discovered that steady-state mRNA levels from transgenes encoding a fully truncated, CTD-less subunit (IIt) are essentially equal to wild type levels, whereas the levels of the CTD-less subunit itself and the amount of polymerase harboring it (Pol IIT) are significantly lower than wild type. In contrast, for the half-CTD mutant (W81), steady-state mRNA levels are somewhat lower than for wild type or IIt, while W81 subunit and polymerase amounts are much less than wild type. Finally, we have tested genetically the ability of CTD mutants to complement (rescue) partially functional RpII215 alleles and have found that IIt fails to complement whereas W81 complements partially to completely. These results suggest that removal of the entire CTD renders polymerase completely defective in vivo, whereas eliminating half of the CTD results in a polymerase with significant in vivo activity. PMID- 7498744 TI - Cellular basis and developmental timing in a size cline of Drosophila melanogaster. AB - We examined 20 Drosophila melanogaster populations collected from a 2600-km north south transect in Australia. In laboratory culture at constant temperature and standard larval density, a genetic cline in thorax length and wing area was found, with both traits increasing with latitude. The cline in wing area was based on clines in both cell size and cell number, but was primarily determined by changes in cell number. Body size and larval development time were not associated among populations. We discuss our results in the context of selection processes operating in natural and experimental populations. PMID- 7498742 TI - A genetic analysis of the Drosophila closely linked interacting genes bulge, argos and soba. AB - The Drosophila gene argos encodes a diffusible protein that acts as a negative regulator of cell fate decisions. To define interacting gene products, we performed a genetic analysis of argos, which suggests the presence of several partially redundant gene functions in its immediate vicinity at the chromosomal position 73A. Dose titration experiments have identified two of these loci. One of them corresponds to the gene bulge. Loss of function bulge alleles suppress the rough eye phenotype associated with overexpression of argos; conversely, amorphic argos mutations suppress the eye phenotype seen in flies bearing a single dominant bulge allele. Recombination mapping localized bulge 0.15 cM distal to argos. A second gene, suppressor of bulge and argos (soba), corresponds to the recently described lethal complementation group 73Aj. soba alleles suppress the eye phenotypes seen in flies expressing either the dominant bulge allele or the hs-argos construct. soba resides 120 kb proximal to argos. In addition, we have identified one allele of a new gene, clown, which like soba suppresses the eye phenotypes associated with hs-argos and bulgeDominant. clown maps on chromosome 3 at the cytological position 68CD. PMID- 7498743 TI - Regulatory regions of the homeotic gene proboscipedia are sensitive to chromosomal pairing. AB - We have identified regulatory regions of the homeotic gene proboscipedia that are capable of repressing a linked white minigene in a manner that is sensitive to chromosomal pairing. Normally, the eye color of transformants containing white in a P-element vector is affected by the number of copies of the transgene; homozygous flies have darker eyes than heterozygotes. However, we found that flies homozygous for select pb DNA-containing transgenes had lighter eyes than heterozygotes. Several pb DNA fragments are capable of causing this pairing sensitive (PS) negative regulation of white. Two fragments in the upstream DNA of pb, 0.58 and 0.98 kb, are PS; additionally, two PS sites are located in the second intron, including a 0.5-kb region and 49-bp sequence. This phenotype is not observed when two PS sites are located at different chromosomal insertion sites (in trans-heterozygous transgenic animals), indicating that the pb-DNA mediated repression of white is dependent on the pairing or proximity of the PS regions. The observed phenomenon is similar to transvection in which certain alleles of a gene can complement each other, but only when homologous chromosomes are paired. Interestingly, the intronic PS regions contain positive regulatory sequences for pb, whereas the upstream PS sites contain pb negative regulatory elements. PMID- 7498746 TI - Microsatellite variation in honey bee (Apis mellifera L.) populations: hierarchical genetic structure and test of the infinite allele and stepwise mutation models. AB - Samples from nine populations belonging to three African (intermissa, scutellata and capensis) and four European (mellifera, ligustica, carnica and cecropia) Apis mellifera subspecies were scored for seven microsatellite loci. A large amount of genetic variation (between seven and 30 alleles per locus) was detected. Average heterozygosity and average number of alleles were significantly higher in African than in European subspecies, in agreement with larger effective population sizes in Africa. Microsatellite analyses confirmed that A. mellifera evolved in three distinct and deeply differentiated lineages previously detected by morphological and mitochondrial DNA studies. Dendrogram analysis of workers from a given population indicated that super-sisters cluster together when using a sufficient number of microsatellite data whereas half-sisters do not. An index of classification was derived to summarize the clustering of different taxonomic levels in large phylogenetic trees based on individual genotypes. Finally, individual population x loci data were used to test the adequacy of the two alternative mutation models, the infinite allele model (IAM) and the stepwise mutation models. The better fit overall of the IAM probably results from the majority of the microsatellites used including repeats of two or three different length motifs (compound microsatellites). PMID- 7498747 TI - A second superoxide dismutase gene in the medfly, Ceratitis capitata. AB - We report the first case of two Cu/Zn Sod genes (ccSod1 and ccSod2) that have been cloned and sequenced from an insect, the medfly, Ceratitis capitata. Biochemical evidence suggested the presence of two Sod genes in the medfly. The two genes are isolated using different molecular strategies: ccSod1 via cross hybridization to a genomic library using a heterologous probe and ccSod2 from cDNA using a homologous probe generated by PCR. Sequence analysis shows that ccSod1 and ccSod2 are different genes. The inferred amino sequences show that all essential residues of the active site are strictly conserved, which suggests both genes encode functional Cu/Zn superoxide dismutase (SOD). Phylogenetic analysis by the maximum parsimony method with bootstrap resampling of previously known Cu/Zn SOD reveals two monophyletic groups, vertebrates and insects. The position of ccSOD2 in this phylogeny is undefined with respect to dipteran ccSOD1, vertebrate, plant, fungal, and extracellular Cu/Zn SOD, which suggests that the duplication detected in Ceratitis is ancient, perhaps as old as the origins of the arthropod phylum in the Cambrian more than 500 million years ago. In situ hybridization to polytene chromosomes places the genes on different chromosomes, which is consistent with an ancient gene duplication. PMID- 7498745 TI - Effects of a transposable element insertion on alcohol dehydrogenase expression in Drosophila melanogaster. AB - Variation in the DNA sequence and level of alcohol dehydrogenase (Adh) gene expression in Drosophila melanogaster have been studied to determine what types of DNA polymorphisms contribute to phenotypic variation in natural populations. The Adh gene, like many others, shows a high level of variability in both DNA sequence and quantitative level of expression. A number of transposable element insertions occur in the Adh region and one of these, a copia insertion in the 5' flanking region, is associated with unusually low Adh expression. To determine whether this insertion (called R142) causes the low expression level, the insertion was excised from the cloned R142 Adh gene and the effect was assessed by P-element transformation. Removal of this insertion causes a threefold increase in the level of ADH, clearly showing that it contributes to the naturally occurring variation in expression at this locus. Removal of all but one LTR also causes a threefold increase, indicating that the mechanism is not a simple sequence disruption. Furthermore, this copia insertion, which is located between the two Adh promoters and their upstream enhancer sequences, has differential effects on the levels of proximal and distal transcripts. Finally, a test for the possible modifying effects of two suppressor loci, su(wa) and su(f), on this insertional mutation was negative, in contrast to a previous report in the literature. PMID- 7498748 TI - An autosomal genetic linkage map of the sheep genome. AB - We report the first extensive ovine genetic linkage map covering 2070 cM of the sheep genome. The map was generated from the linkage analysis of 246 polymorphic markers, in nine three-generation full-sib pedigrees, which make up the AgResearch International Mapping Flock. We have exploited many markers from cattle so that valuable comparisons between these two ruminant linkage maps can be made. The markers, used in the segregation analyses, comprised 86 anonymous microsatellite markers derived from the sheep genome, 126 anonymous microsatellites from cattle, one from deer, and 33 polymorphic markers of various types associated with known genes. The maximum number of informative meioses within the mapping flock was 222. The average number of informative meioses per marker was 140 (range 18-209). Linkage groups have been assigned to all 26 sheep autosomes. PMID- 7498749 TI - Radiation-sensitive mutants of Arabidopsis thaliana. AB - Five Arabidopsis mutants have been isolated on the basis of hypersensitivity of leaf tissue to UV light. For each mutant, the UV-hypersensitive phenotype (uvh) was inherited as a single recessive Mendelian trait. In addition, each uvh mutant represented a separate complementation group. Three of the mutations producing the UV hypersensitive phenotype have been mapped relative to either genetic markers or physical microsatellite polymorphisms. Locus UVH1 is linked to nga76 on chromosome 5, UVH3 to GL1 on chromosome three, and UVH6 to nga59 on chromosome 1. Each uvh mutant has a characteristic pattern of sensitivity based on UV sensitivity of leaf tissue, UV sensitivity of root tissue, and ionizing radiation sensitivity of seeds. On the basis of these patterns, possible molecular defects in these mutants are discussed. PMID- 7498750 TI - Population structure of morphological traits in Clarkia dudleyana. I. Comparison of FST between allozymes and morphological traits. AB - Studies of genetic variation at allozyme loci, assumed to be selectively neutral, have provided valuable insights into the genetic structure of numerous populations. The degree to which population structure of allozyme variation reflects that of quantitative traits, however, is not well resolved. Here, we compare estimates of population differentiation (FST) of 11 populations for allozymes with those for nine discrete and nine continuous morphological traits. Overall, the allozymes have the lowest FST estimates, indicating relatively little population differentiation. Excepting two traits, petal width and long internode length, the continuous morphological traits have estimates similar to those from allozymes. The discrete morphological traits tend to have the highest estimates. On a single trait basis, estimates of FST for four discrete and two continuous traits are higher than those for allozymes. A more detailed (narrow sense quantitative) genetic study of two populations suggests that these estimates of FST may underestimate the true value because of dominance. Clustering analyses show that the pattern of differentiation for the discrete morphological traits strongly reflects the geographical distribution of the populations, whereas the patterns for the continuous traits and allozymes do not. These results suggest that selection has been occurring on the discrete morphological traits, selecting toward a common optimum within each geographic group, and optima differing among geographic groups. PMID- 7498751 TI - Dominance is the major genetic basis of heterosis in rice as revealed by QTL analysis using molecular markers. AB - A set of 194 F7 lines derived from a subspecific rice cross showing strong F1 heterosis was backcrossed to the two parents. The materials (388 BC1F7 lines, 194 F8 lines, two parents, F1) were phenotyped for 12 quantitative traits. A total of 37 significant QTLs (LOD > or = 2.0) was detected through 141 RFLP markers in the BC1F7 populations. Twenty-seven (73%) quantitative trait loci (QTLs) were detected in only one of the BC1F7 populations. In 82% of these cases, the heterozygotes were superior to the respective homozygotes. The remaining 10 (27%) QTLs were detected in both BC1F7 populations, and the heterozygote had a phenotype falling between those of the two homozygotes and in no instances were the heterozygotes found to be superior to both homozygotes. These results suggest that dominance complementation is the major genetic basis of heterosis in rice. This conclusion was strengthened by the finding that there was no correlation between most traits and overall genome heterozygosity and that there were some recombinant inbred lines in the F8 population having phenotypic values superior to the F1 for all of the traits evaluated--a result not expected if overdominance was a major contributor to heterosis. Digenic epistasis was not evident. PMID- 7498752 TI - Heterozygosity and fitness: no association in Scots pine. AB - The association of six quantitative traits related to fitness with heterozygosity at 12 allozyme loci has been examined in three populations of Scots pine, Pinus sylvestris. Because of several characteristics of this organism and of this extensive data set, it appeared that this study would show a positive association between heterozygosity and these traits if indeed heterozygotes had higher values for these quantitative traits. Using several different statistical techniques including analysis of variance, regression with the scaling recommended from the adaptive distance model, and multiple regression, no evidence of an association was found. For example, only between 7 and 8% of the regression tests were significant at the 5% level and half of these showed a positive association and half showed a negative association. Further, the multiple regression analysis explained on average only 5.8% of the variation observed in the six different traits and only 1.5% of this variation was explained by a positive association. Power analysis was carried out (for the first time on these type of data), both for the single locus heterozygous advantage and the association of individual multiple locus heterozygosity and the quantitative traits. For diameter and height, two traits often used in similar studies, the average power to detect a single locus heterozygous advantage of 0.10 was 0.737 and the average power to detect a mean heterozygote advantage of 0.05 per locus for multiple loci was 0.797. As a result of this study and an examination of the published results from other studies, it appears that what positive associations have been observed are probably not, in large part, due to the presence of intrinsic heterozygote advantage. PMID- 7498753 TI - Separating population structure from population history: a cladistic analysis of the geographical distribution of mitochondrial DNA haplotypes in the tiger salamander, Ambystoma tigrinum. AB - Nonrandom associations of alleles or haplotypes with geographical location can arise from restricted gene flow, historical events (fragmentation, range expansion, colonization), or any mixture of these factors. In this paper, we show how a nested cladistic analysis of geographical distances can be used to test the null hypothesis of no geographical association of haplotypes, test the hypothesis that significant associations are due to restricted gene flow, and identify patterns of significant association that are due to historical events. In this last case, criteria are given to discriminate among contiguous range expansion, long-distance colonization, and population fragmentation. The ability to make these discriminations depends critically upon an adequate geographical sampling design. These points are illustrated with a worked example: mitochondrial DNA haplotypes in the salamander Ambystoma tigrinum. For this example, prior information exists about restricted gene flow and likely historical events, and the nested cladistic analyses were completely concordant with this prior information. This concordance establishes the plausibility of this nested cladistic approach, but much future work will be necessary to demonstrate robustness and to explore the power and accuracy of this procedure. PMID- 7498754 TI - The hitchhiking effect on the site frequency spectrum of DNA polymorphisms. AB - The level of DNA sequence variation is reduced in regions of the Drosophila melanogaster genome where the rate of crossing over per physical distance is also reduced. This observation has been interpreted as support for the simple model of genetic hitchhiking, in which directional selection on rare variants, e.g., newly arising advantageous mutants, sweeps linked neutral alleles to fixation, thus eliminating polymorphisms near the selected site. However, the frequency spectra of segregating sites of several loci from some populations exhibiting reduced levels of nucleotide diversity and reduced numbers of segregating sites did not appear different from what would be expected under a neutral equilibrium model. Specifically, a skew toward an excess of rare sites was not observed in these samples, as measured by Tajima's D. Because this skew was predicted by a simple hitchhiking model, yet it had never been expressed quantitatively and compared directly to DNA polymorphism data, this paper investigates the hitchhiking effect on the site frequency spectrum, as measured by Tajima's D and several other statistics, using a computer simulation model based on the coalescent process and recurrent hitchhiking events. The results presented here demonstrate that under the simple hitchhiking model (1) the expected value of Tajima's D is large and negative (indicating a skew toward rare variants), (2) that Tajima's test has reasonable power to detect a skew in the frequency spectrum for parameters comparable to those from actual data sets, and (3) that the Tajima's Ds observed in several data sets are very unlikely to have been the result of simple hitchhiking. Consequently, the simple hitchhiking model is not a sufficient explanation for the DNA polymorphism at those loci exhibiting a decreased number of segregating sites yet not exhibiting a skew in the frequency spectrum. PMID- 7498755 TI - Dependence of frequency of homologous recombination on the homology length. AB - The frequency of homologous recombination is believed to be a linear function of the length (N bp) of homology between DNAs. Here, the N intercept is believed to be determined by a threshold length below which some physical constraint is effective. In the mammalian gene targeting systems, however, the frequency depends more steeply than linearly on the homology length. To explain both the linear dependence and the steeper dependence, we propose a model where the branch point of a reaction intermediate is assumed to "walk randomly" along the homologous region until it is processed. The intermediate is assumed to be destroyed if the branch point ever reaches either end of the homology. In this model, the length dependence is governed by a parameter, h, which is defined as efficiency of processing of the intermediate and reflects unlikelihood of the destruction at either end of the homology. We find that the frequency is proportional to N3 for smaller N and is a linear function of N for larger N. Where the shift from the N3 dependence to the linear dependence takes place is determined by the parameter h. The range of N showing the N3 dependence becomes narrower as h becomes larger. The dependence steeper than linear dependence, which is observed not only in the mammalian gene targeting system but also in bacteriophage T4, Escherichia coli and yeast systems, agrees well with the predicted N3 dependence. The N intercept is determined not by physical (or structural) constraints but only by the parameter h in this model. PMID- 7498756 TI - Analysis of DNA diversity by spatial autocorrelation. AB - Two statistics are proposed for summarizing spatial patterns of DNA diversity. These autocorrelation indices for DNA analysis, or AIDAs, can be applied to RFLP and sequence data; the resulting set of autocorrelation coefficients, or correlogram, measures whether, and to what extent, individual DNA sequences or haplotypes resemble the haplotypes sampled at arbitrarily chosen spatial distances. Analyses of computer-generated sets of data, and of RFLP data from two natural populations, show that AIDAs allow one to objectively and simply identify basic patterns in the spatial distribution of haplotypes. These statistics, therefore, seem to be a useful tool both to explore the genetic structure of a population and to suggest hypotheses on the evolutionary processes that shaped the observed patterns. PMID- 7498757 TI - Linkage and the limits to natural selection. AB - The probability of fixation of a favorable mutation is reduced if selection at other loci causes inherited variation in fitness. A general method for calculating the fixation probability of an allele that can find itself in a variety of genetic backgrounds is applied to find the effect of substitutions, fluctuating polymorphisms, and deleterious mutations in a large population. With loose linkage, r, the effects depend on the additive genetic variance in relative fitness, var(W), and act by reducing effective population size by (N/Ne) = 1 + var(W)/2r2. However, tightly linked loci can have a substantial effect not predictable from Ne. Linked deleterious mutations reduce the fixation probability of weakly favored alleles by exp (-2U/R), where U is the total mutation rate and R is the map length in Morgans. Substitutions can cause a greater reduction: an allele with advantage s < scrit = (pi 2/6) loge (S/s) [var(W)/R] is very unlikely to be fixed. (S is the advantage of the substitution impeding fixation.) Fluctuating polymorphisms at many (n) linked loci can also have a substantial effect, reducing fixation probability by exp [square root of 2Kn var(W)/R] [K = 1/E((u-u)2/uv) depending on the frequencies (u,v) at the selected polymorphisms]. Hitchhiking due to all three kinds of selection may substantially impede adaptation that depends on weakly favored alleles. PMID- 7498758 TI - Statistical methods for linkage analysis of complex traits from high-resolution maps of identity by descent. AB - A multilocus model for complex traits is described that generalizes the additive and multiplicative models and hence allows simultaneously for both heterogeneity and gene interaction (epistasis). Statistical methods of linkage analysis are discussed under the assumption that identity by descent data from a dense set of polymorphic markers are available. Three methods, single locus search, simultaneous search and conditional search, are described and compared. PMID- 7498759 TI - Two genes, no enzyme: a second look at Barbara McClintock and the 1951 Cold Spring Harbor Symposium. PMID- 7498760 TI - Host and phi X 174 mutations affecting the morphogenesis or stabilization of the 50S complex, a single-stranded DNA synthesizing intermediate. AB - The morphogenetic pathway of bacteriophage phi X 174 was investigated in rep mutant hosts that specifically block stage III single-stranded DNA synthesis. The defects conferred by the mutant rep protein most likely affect the formation or stabilization of the 50S complex, a single-stranded DNA synthesizing intermediate, which consists of a viral prohead and a DNA replicating intermediate (preinitiation complex). phi X 174 mutants, ogr (rep), which restore the ability to propagate in the mutant rep hosts, were isolated. The org (rep) mutations confer amino acid substitutions in the viral coat protein, a constituent of the prohead, and the viral A protein, a constituent of the preinitiation complex. Four of the six coat protein substitutions are localized on or near the twofold axis of symmetry in the atomic structure of the mature virion. PMID- 7498761 TI - DNA structures generated during recombination initiated by mismatch repair of UV irradiated nonreplicating phage DNA in Escherichia coli: requirements for helicase, exonucleases, and RecF and RecBCD functions. AB - During infection of homoimmune Escherichia coli lysogens ("repressed infections"), undamaged nonreplicating lambda phage DNA circles undergo very little recombination. Prior UV irradiation of phages dramatically elevates recombinant frequencies, even in bacteria deficient in UvrABC-mediated excision repair. We previously reported that 80-90% of this UvrABC-independent recombination required MutHLS function and unmethylated d(GATC) sites, two hallmarks of methyl-directed mismatch repair. We now find that deficiencies in other mismatch-repair activities--UvrD helicase, exonuclease I, exonuclease VII, RecJ exonuclease--drastically reduce recombination. These effects of exonuclease deficiencies on recombination are greater than previously observed effects on mispair-provoked excision in vitro. This suggests that the exonucleases also play other roles in generation and processing of recombinagenic DNA structures. Even though dsDNA breaks are thought to be highly recombinagenic, 60% of intracellular UV-irradiated phage DNA extracted from bacteria in which recombination is low- UvrD-, ExoI-, ExoVII-, or Rec(J-)--displays (near-)blunt-ended dsDNA ends (RecBCD sensitive when deproteinized). In contrast, only bacteria showing high recombination (Mut+ UvrD+ Exo+) generate single-stranded regions in nonreplicating UV-irradiated DNA. Both recF and recB recC mutations strikingly reduce recombination (almost as much as a recF recB recC triple mutation), suggesting critical requirements for both RecF and RecBCD activity. The mismatch repair system may thus process UV-irradiated DNA so as to initiate more than one recombination pathway. PMID- 7498762 TI - The diversity of alleles at the hsd locus in natural populations of Escherichia coli. AB - In enteric bacteria three discrete families of type I restriction and modification systems (IA, IB and ID) are encoded by alleles of the serB-linked hsd locus. Probes specific for each of the three families were used to monitor the distribution of related systems in 37 of the 72 wild-type Escherichia coli strains comprising the ECOR collection. All 25 members of group A in this collection were screened; 12 were probe-positive, nine have hsd genes in the IA family, two in the IB and one in the ID. Twelve strains, representing all groups other than A, were screened; five were probe-positive, one has hsd genes in the IA family, one in the IB and three in the ID. The type ID genes are the first representatives of this family in E. coli, the probe-negative strains could have alternative families of hsd genes. The type IA and IB systems added at least five new specificities to the five already identified in natural isolates of E. coli. The distribution of alleles is inconsistent with the dendrogram of the bacterial strains derived from other criteria. This discrepancy and the dissimilar coding sequences of allelic hsd genes both imply lateral transfer of hsd genes. PMID- 7498764 TI - Genetics of the synthesis of serine from glycine and the utilization of glycine as sole nitrogen source by Saccharomyces cerevisiae. AB - Saccharomyces cerevisiae can grow on glycine as sole nitrogen source and can convert glycine to serine via the reaction catalyzed by the glycine decarboxylase multienzyme complex (GDC). Yeast strains with mutations in the single gene for lipoamide dehydrogenase (lpd1) lack GDC activity, as well as the other three 2 oxoacid dehydrogenases dependent on this enzyme. The LPD1 gene product is also required for cells to utilize glycine as sole nitrogen source. The effect of mutations in LPD1 (L-subunit of GDC), SER1 (synthesis of serine from 3 phosphoglycerate), ADE3 (cytoplasmic synthesis of one-carbon units for the serine synthesis from glycine), and all combinations of each has been determined. The results were used to devise methods for isolating mutants affected either in the generation of one-carbon units from glycine (via GDC) or subsequent steps in serine biosynthesis. The mutants fell into six complementation groups (gsd1-6 for defects in conversion of glycine to serine). Representatives from three complementation groups were also unable to grow on glycine as sole nitrogen source (gsd1-3). Assays of the rate of glycine uptake and decarboxylation have provided insights into the nature of the mutations. PMID- 7498763 TI - The role of DNA repair genes in recombination between repeated sequences in yeast. AB - The presence of repeated sequences in the genome represents a potential source of karyotypic instability. Genetic control of recombination is thus important to preserve the integrity of the genome. To investigate the genetic control of recombination between repeated sequences, we have created a series of isogenic strains in which we could assess the role of genes involved in DNA repair in two types of recombination: direct repeat recombination and ectopic gene conversion. Naturally occurring (Ty elements) and artificially constructed repeats could be compared in the same cell population. We have found that direct repeat recombination and gene conversion have different genetic requirements. The role of the RAD51, RAD52, RAD54, RAD55, and RAD57 genes, which are involved in recombinational repair, was investigated. Based on the phenotypes of single and double mutants, these genes can be divided into three functional subgroups: one composed of RAD52, a second one composed of RAD51 and RAD54, and a third one that includes the RAD55 and RAD57 genes. Among seven genes involved in excision repair tested, only RAD1 and RAD10 played a role in the types of recombination studied. We did not detect a differential effect of any rad mutation on Ty elements as compared to artificially constructed repeats. PMID- 7498765 TI - Construction and analysis of yeast RNA polymerase II CTD deletion and substitution mutations. AB - The carboxyl-terminal domain (CTD) of the RNA polymerase II largest subunit plays an essential but poorly understood role in transcription. The CTD is highly phosphorylated in vivo and this modification may be important in the transition from transcription initiation to elongation. We report here the development of a strategy for creating novel yeast CTDs. We have used this approach to show that the minimum viable CTD in yeast contains eight consensus (Tyr1Ser2Pro3Thr4Ser5Pro6Ser7) heptapeptide repeats. Substitution of alanine or glutamate for serines in positions two or five is lethal. In addition, changing tyrosine in position one to phenylalanine is lethal. The effects of mutations that alter potential phosphorylation sites are consistent with a requirement for CTD phosphorylation in vivo. PMID- 7498766 TI - The role of cdc2 and other genes in meiosis in Schizosaccharomyces pombe. AB - The requirement of the cdc2, cdc13 and cdc25 genes for meiosis in Schizosaccharomyces pombe was investigated using three different conditions to induce meiosis. These genes were known to be required for meiosis II. cdc13 and cdc25 are essential for meiosis I. The cdc2 gene, which is required for the initiation of both mitotic S-phase and M-phase, is essential for premeiotic DNA synthesis and meiosis II. The requirement of cdc2 for meiosis I was unclear. This contrasts with Saccharomyces cerevisiae, where CDC28, the homolog of cdc2, is required for meiosis I but not for premeiotic DNA synthesis. Expression of cdc13 and cdc25 was induced after premeiotic DNA synthesis, reaching a sharp peak before the first nuclear division. Expression of cdc22, encoding the large subunit of ribonucleotide reductase, was also induced but the peak was before premeiotic DNA synthesis. The induction of cdc13 and cdc25 was largely dependent on DNA synthesis and the function of the mei4 gene. The mei4 gene itself was also induced in a DNA synthesis-dependent manner. The chain of gene expression activating cdc25 may be important as part of the mechanism that ensures the dependency of nuclear division on DNA replication during meiosis. PMID- 7498767 TI - Alterations in ribosomal protein RPS28 can diversely affect translational accuracy in Saccharomyces cerevisiae. AB - Three small-subunit ribosomal proteins shown to influence translational accuracy in Saccharomyces cerevisiae are conserved in structure and function with their procaryotic counterparts. One of these, encoded by RPS28A and RPS28B (RPS28), is comparable to bacterial S12. The others, encoded by sup44 (RPS4) or, sup46 and YS11A (RPS13), are homologues of procaryotic S5 and S4, respectively. In Escherichia coli, certain alterations in S12 cause hyperaccurate translation or antibiotic resistance that can be counteracted by other changes in S5 or S4 that reduce translational accuracy. Using site-directed and random mutagenesis, we show that different changes in RPS28 can have diametrical influences on translational accuracy or antibiotic sensitivity in yeast. Certain substitutions in the amino-terminal portion of the protein, which is diverged from the procaryotic homologues, cause varying levels of nonsense suppression or antibiotic sensitivity. Other alterations, found in the more conserved carboxyl terminal portion, counteract SUP44- or SUP46-associated antibiotic sensitivity, mimicking E. coli results. Although mutations in these different parts of RPS28 have opposite affects on translational accuracy or antibiotic sensitivity, additive phenotypes can be observed when opposing mutations are combined in the same protein. PMID- 7498768 TI - Mutational analysis of morphologic differentiation in Saccharomyces cerevisiae. AB - A genetic analysis was undertaken to investigate the mechanisms controlling cellular morphogenesis in Saccharomyces cerevisiae. Sixty mutant strains exhibiting abnormally elongated cell morphology were isolated. The cell elongation phenotype in at least 26 of the strains resulted from a single recessive mutation. These mutations, designated generically elm (elongated morphology), defined 14 genes; two of these corresponded to the previously described genes GRR1 and CDC12. Genetic interactions between mutant alleles suggest that several ELM genes play roles in the same physiological process. The cell and colony morphology and growth properties of many elm mutant strains are similar to those of wild-type yeast strains after differentiation in response to nitrogen limitation into the pseudohyphal form. Each elm mutation resulted in multiple characteristics of pseudohyphal cells, including elongated cell shape, delay in cell separation, simultaneous budding of mother and daughter cells, a unipolar budding pattern, and/or the ability to grow invasively beneath the agar surface. Mutations in 11 of the 14 ELM gene loci potentiated pseudohyphal differentiation in nitrogen-limited medium. Thus, a subset of the ELM genes are likely to affect control or execution of a defined morphologic differentiation pathway in S. cerevisiae. PMID- 7498770 TI - Drosophila-host genetic control of susceptibility to Drosophila C virus. AB - Interactions between Drosophila C virus (DCV) and its natural host, Drosophila melanogaster, were investigated using 15 geographical population samples infected by intraabdominal inoculation. These strains derived from natural populations of D. melanogaster differed in susceptibility to the DCVc. One strain was "partially tolerant". Isofemale lines obtained from one susceptible and one partially tolerant strain were studied. The partially tolerant phenotype was dominant, and there was no difference between F1 progeny of direct and reciprocal crosses. Analysis of F2 progeny showed that neither sex-linked genes nor maternal effects are involved in susceptibility to DCVc. The partially tolerant strain phenotype was dominant and segregated with chromosome III. Two nonexclusive hypotheses are proposed to explain chromosome III gene action. PMID- 7498769 TI - Non-locus-specific polygenes giving responses to selection for gene conversion frequencies in Ascobolus immersus. AB - Selection for higher and lower meiotic conversion frequencies was investigated in the fungus Ascobolus immersus. Strains carrying the same known gene conversion control factors, which have major effects on conversion frequencies at their specific target locus, sometimes gave significant differences in conversion frequency. Selection for high or low conversion frequencies at the w1-78 site was practiced for five generations, giving significant responses in both directions. These responses were due to polygenes, or genes of minor effect, not to new conversion control factors of major effect. Crosses of selected strains to strains with other mutations showed that the genes' effects were not specific to w1-78, but could affect conversion frequencies of another mutation, w1-3C1, at that locus and of two other loci, w-BHj and w9, which are unlinked to w1 or to each other. The proportional changes in gene conversion frequency due to selection varied according to the locus and site involved and according to the conversion control factor alleles present. There were differences of > or = 277% in conversion frequency between "high" and "low" strains. Selection for conversion frequency had little effect on other features of conversion, such as the frequency of postmeiotic segregation or the relative frequencies of conversion to mutant or wild type. PMID- 7498771 TI - Colonization of the Americas by Drosophila subobscura: lethal-gene allelism and association with chromosomal arrangements. AB - Drosophila subobscura is a Palearctic species that has recently colonized the Americas. It was first found in 1978 in Puerto Montt, Chile, and in 1982 in Port Townsend, WA. The colonization and rapid expansion of the species in western South and North America provides distinctive opportunities for investigating the process of evolution in action. The inversion polymorphism in the O chromosome from populations of central California and northern Washington, separated by 1300 km, corresponds to a previously observed latitudinal cline, also observed in Europe. Recessive lethal genes are not randomly distributed among the chromosomal arrangements. The incidence of lethal allelism is high, yielding unrealistically low estimates of the effective size of these populations (on the order of 1000 individuals). The high incidence of lethal allelism is likely to be a consequence of the low number of the American colonizers (on the order of 10-100 individuals), but the persistence of the allelism over several years suggests that some lethal-carrying chromosomes may be heterotic owing to shared associations between lethal and other genes. PMID- 7498772 TI - Wolbachia infections and the expression of cytoplasmic incompatibility in Drosophila sechellia and D. mauritiana. AB - Various stocks of Drosophila mauritiana and D. sechellia were found to be infected with Wolbachia, a Rickettsia-like bacterium that is known to cause cytoplasmic incompatibility and other reproductive abnormalities in arthropods. Testing for the expression of cytoplasmic incompatibility in these two species showed partial incompatibility in D. sechellia but no expression of incompatibility in D. mauritiana. To determine whether absence of cytoplasmic incompatibility in D. mauritiana was due to either the bacterial or host genome, we transferred bacteria from D. mauritiana into an uninfected strain of D. simulans, a host species known to express high levels of incompatibility with endogenous Wolbachia. We also performed the reciprocal transfer of the natural D. simulans Riverside infection into a tetracycline-treated stock of D. mauritiana. In each case, the ability to express incompatibility was unaltered by the different host genetic background. These experiments indicate that in D. simulans and D. mauritiana expression of the cytoplasmic incompatibility phenotype is determined by the bacterial strain and that D. mauritiana harbors a neutral strain of Wolbachia. PMID- 7498773 TI - Cytoplasmic incompatibility in Drosophila simulans: dynamics and parameter estimates from natural populations. AB - In Drosophila simulans, cytoplasmically transmitted Wolbachia microbes cause reduced egg hatch when infected males mate with uninfected females. A Wolbachia infection and an associated mtDNA variant have spread northward through California since 1986. PCR assays show that Wolbachia infection is prevalent throughout the continental US and Central and South America, but some lines from Florida and Ecuador that are PCR-positive for Wolbachia do not cause incompatibility. We estimate from natural populations infection frequencies and the transmission and incompatibility parameter values that affect the spread of the infection. On average, infected females from nature produce 3-4% uninfected ova. Infected females with relatively low fidelity of maternal transmission show partial incompatibility with very young infected laboratory males. Nevertheless, crosses between infected flies in nature produce egg-hatch rates indistinguishable from those produced by crosses between uninfected individuals. Incompatible crosses in nature produce hatch rates 30-70% as high as those from compatible crosses. Wild-caught infected and uninfected females are equally fecund in the laboratory. Incompatibility decreases with male age, and age specific incompatibility levels suggest that males mating in nature may often be 2 or 3 weeks old. Our parameter estimates accurately predict the frequency of Wolbachia infection in California populations. PMID- 7498774 TI - A screen for modifiers of Deformed function in Drosophila. AB - Proteins produced by the homeotic genes of the Hox family assign different identifies to cells on the anterior/posterior axis. Relatively little is known about the signalling pathways that modulate their activities or the factors with which they interact to assign specific segmental identifies. To identify genes that might encode such functions, we performed a screen for second site mutations that reduce the viability of animals carrying hypomorphic mutant alleles of the Drosophila homeotic locus, Deformed. Genes mapping to six complementation groups on the third chromosome were isolated as modifiers of Deformed function. Products of two of these genes, sallimus and moira, have been previously proposed as homeotic activators since they suppress the dominant adult phenotype of Polycomb mutants. Mutations in hedgehog, which encodes secreted signalling proteins, were also isolated as Deformed loss-of-function enhancers. Hedgehog mutant alleles also suppress the Polycomb phenotype. Mutations were also isolated in a few genes that interact with Deformed but not with Polycomb, indicating that the screen identified genes that are not general homeotic activators. Two of these genes, cap 'n' collar and defaced, have defects in embryonic head development that are similar to defects seen in loss of function Deformed mutants. PMID- 7498775 TI - Complete sequence and gene organization of the mitochondrial genome of the land snail Albinaria coerulea. AB - The complete sequence (14,130 bp) of the mitochondrial DNA (mtDNA) of the land snail Albinaria coerulea was determined. It contains 13 protein, two rRNA and 22 tRNA genes. Twenty-four of these genes are encoded by one and 13 genes by the other strand. The gene arrangement shares almost no similarities with that of two other molluscs for which the complete gene content and arrangement are known, the bivalve Mytilus edulis and the chiton Katharina tunicata; the protein and rRNA gene order is similar to that of another terrestrial gastropod, Cepaea nemoralis. Unusual features include the following: (1) the absence of lengthy noncoding regions (there are only 141 intergenic nucleotides interspersed at different gene borders, the longest intergenic sequence being 42 nucleotides) (2) the presence of several overlapping genes (mostly tRNAs), (3) the presence of tRNA-like structures and other stem and loop structures within genes. An RNA editing system acting on tRNAs must necessarily be invoked for posttranscriptional extension of the overlapping tRNAs. Due to these features, and also because of the small size of its genes (e.g., it contains the smallest rRNA genes among the known coelomates), it is one of the most compact mitochondrial genomes known to date. PMID- 7498776 TI - Ribosomal RNA multigene loci: nomads of the Triticeae genomes. AB - The nucleolus organizing regions (NORs) on the short arms of chromosomes 1A(m) and 5A(m) of diploid wheat, Triticum monococcum L., are at the most distal loci in the linkage maps of these two chromosome arms. This distal location differs from the interstitial location of the Nor loci on chromosome arms 1BS of tetraploid Triticum turgidum L. and hexaploid T. aestivum L., 5DS of T. aestivum and diploid Ae. tauschii Coss., and 5HS of barley. Moreover, the barley 5HS locus is at a different location than the 5DS locus. However, other markers, including the centromeres, are colinear. These findings showed that the major Nor loci have repeatedly changed position in the chromosome arms during the radiation of species in the tribe Triticeae without rearrangements of the linkage groups. It is suggested that Nor loci may change position via dispersion of minor loci, that are shown here to exist in the T. monococcum genome, magnification of gene copy numbers in these minor loci, and subsequent deletion of the original major loci. Implications of these findings for the use of rRNA nucleotide sequences in phylogenetic reconstructions are pointed out. PMID- 7498777 TI - Environmental programming of heritable epigenetic changes in paramutant r-gene expression using temperature and light at a specific stage of early development in maize seedlings. AB - Different heritable expression-states were programmed into R alleles from R/R-1st heterozygotes under different temperature conditions applied during a developmental period in which flowering is induced. At maturity, R-allele expressions in test crosses of male gametes derived from R/R-1st seedlings raised 15 days in 32 degrees and continuous light conditions differed significantly from those of sib seedlings raised for 15 days in 22 degrees and continuous light conditions and shifted to six 12-hr light-dark cycles, day 16-21. This experiment provides the first evidence in higher organisms that environmental conditions, applied at a specific stage of development cause a heritable change in a specific allele expression. My earlier evidence required a statistical analysis for demonstrating heritable change; I present photographic evidence of this environmental effect on four R alleles. PMID- 7498779 TI - Synonymous substitution rates in enterobacteria. AB - It has been shown previously that the synonymous substitution rate between Escherichia coli and Salmonella typhimurium is lower in highly than in weakly expressed genes, and it has been suggested that this is due to stronger selection for translational efficiency in highly expressed genes as reflected in their greater codon usage bias. This hypothesis is tested here by comparing the substitution rate in codon families with different patterns of synonymous codon use. It is shown that the decline in the substitution rate across expression levels is as great for codon families that do not appear to be subject to selection for translational efficiency as for those that are. This implies that selection on translational efficiency is not responsible for the decline in the substitution rate across genes. It is argued that the most likely explanation for this decline is a decrease in the mutation rate. It is also shown that a simple evolutionary model in which synonymous codon use is determined by a balance between mutation, selection for an optimal codon, and genetic drift predicts that selection should have little effect on the substitution rate in the present case. PMID- 7498778 TI - Sequences required for paramutation of the maize b gene map to a region containing the promoter and upstream sequences. AB - The b gene encodes a transcriptional regulator of the maize anthocyanin biosynthetic pathway. Certain b alleles participate in paramutation, an allele specific interaction that heritably alters transcription. The moderately transcribed B' allele heritably reduces the transcription of the highly transcribed B-I allele in a B'/B-I heterozygote, such that the B-I allele becomes B'. To identify the cis-acting sequences required for paramutation, we used B' or B-I alleles to isolate intragenic recombinants with B-Peru, an allele that is insensitive to paramutation and has distinct tissue-specific regulation. Physical mapping of the recombinant alleles showed that most of the crossovers were in a small region near the 5' end of the b-transcribed region. Analysis of the recombinant alleles revealed that the ability to cause and respond to paramutation and the control of tissue-specific expression both localize to the 5' region of the gene. The 3' boundary of these functions lies just upstream of the translation initiation codon. The 5' boundary has been estimated to be no more than 0.1 cM further upstream (1-150 kb). Thus, sequences critical for paramutation lie upstream of the b coding sequences and may include transcriptional regulatory sequences. PMID- 7498780 TI - Testing heterozygote excess and deficiency. AB - Currently used tests of Hardy-Weinberg proportions do not take into account the nature of the alternative hypothesis, which is generally a heterozygote deficiency. Different exact tests, appropriate for small sample size and large number of alleles, are proposed in this perspective, and their properties are evaluated by power comparisons. Some tests are found to be close to optimal for the detection of inbreeding or heterozygote excess, one of which is a score test closely related to Robertson and Hill's estimator of the inbreeding coefficient. This test is also easily applied to multiple samples. Such tests are not always the most appropriate if alternative hypotheses differ from those considered here. PMID- 7498781 TI - Estimating effective population size and mutation rate from sequence data using Metropolis-Hastings sampling. AB - We present a new way to make a maximum likelihood estimate of the parameter 4N mu (effective population size times mutation rate per site, or theta) based on a population sample of molecular sequences. We use a Metropolis-Hastings Markov chain Monte Carlo method to sample genealogies in proportion to the product of their likelihood with respect to the data and their prior probability with respect to a coalescent distribution. A specific value of theta must be chosen to generate the coalescent distribution, but the resulting trees can be used to evaluate the likelihood at other values of theta, generating a likelihood curve. This procedure concentrates sampling on those genealogies that contribute most of the likelihood, allowing estimation of meaningful likelihood curves based on relatively small samples. The method can potentially be extended to cases involving varying population size, recombination, and migration. PMID- 7498782 TI - Spatial distributions of genotypes under isolation by distance. AB - The spatial distributions of single-locus diploid genotypes, produced within populations of sexually reproducing individuals under isolation by distance, are measured and characterized in detail by quantifying the join-counts for simulated model populations. The models more realistically reflect spatially explicit distributions of genotypes in populations, because unlike the classical theory, they include the stochasticity inherent in the process of matings between genotypes. This stochasticity causes the formation of large areas or patches containing mostly one homozygous genotype, which is not predicted by the classical theory. A number of previously uncharacterized features of the spatial structures produced under isolation by distance are revealed. Spatial autocorrelation measures based on counts of pairs of homozygotes are highly consistent quantifications of the concentrations of homozygotes in patchy genotypic distributions for a given level of dispersal. Most strikingly, the degree of intermixing of homozygotes with heterozygotes over small spatial scales is much higher than previously thought, unless dispersal is extremely limited (e.g., Wright's neighborhood size less than approximately 5.0). Among implications for studies of spatial structure of real populations, one is that the results provide a basis for using join-counts as estimators of gene dispersal based solely on genetic data. PMID- 7498783 TI - The complexity of the interaction between RAD52 and SRS2. PMID- 7498784 TI - Approximate variance of nucleotide divergence between two sequences estimated from restriction fragment data. PMID- 7498785 TI - E2F-1 accumulation bypasses a G1 arrest resulting from the inhibition of G1 cyclin-dependent kinase activity. AB - Numerous experiments have defined a critical role for the G1 cyclins and associated kinases in allowing a normal progression of cells from a quiescent state, through G1, and into S phase. We now demonstrate that G1 cyclin-dependent kinase activity is critical for the accumulation of E2F activity late in G1. Moreover, E2F-1 overexpression can overcome a G1 arrest caused by the inhibition of G1 cyclin-dependent kinase activity, consistent with E2F activation being an important consequence of the action of G1 cyclins. E2F-1 also overcomes a G1 block caused by gamma irradiation and leads to an apparent complete replication of the cellular genome and entry into mitosis. This E2F-1-mediated induction of S phase and mitosis is not accompanied by the rise in either cyclin D-associated kinase activity or cdk2 activity that is normally observed during the G1 phase of the cell cycle. We conclude that one key function for G1 cyclin-dependent kinase activity is the activation of E2F-1, that the accumulation of E2F activity may be sufficient to allow initiation and completion of S phase, but that additional events, including G1 cyclin kinase activity, are likely necessary for a normal proliferative event. PMID- 7498786 TI - The SIR2 gene family, conserved from bacteria to humans, functions in silencing, cell cycle progression, and chromosome stability. AB - Genomic silencing is a fundamental mechanism of transcriptional regulation, yet little is known about conserved mechanisms of silencing. We report here the discovery of four Saccharomyces cerevisiae homologs of the SIR2 silencing gene (HSTs), as well as conservation of this gene family from bacteria to mammals. At least three HST genes can function in silencing; HST1 overexpression restores transcriptional silencing to a sir2 mutant and hst3 hst4 double mutants are defective in telomeric silencing. In addition, HST3 and HST4 together contribute to proper cell cycle progression, radiation resistance, and genomic stability, establishing new connections between silencing and these fundamental cellular processes. PMID- 7498787 TI - The tetratricopeptide repeats of Ssn6 interact with the homeo domain of alpha 2. AB - The tetratricopeptide repeat (TPR) is a 34-amino-acid degenerate sequence motif that is found in a large variety of proteins, both prokaryotic and eukaryotic. TPRs are usually found in tandem arrays of up to 16 copies. In this paper we identify a direct interaction between the TPRs of Ssn6, a general transcriptional repressor, and alpha 2, a cell-type regulator in Saccharomyces cerevisiae. Five of the Ssn6 TPRs were tested individually, and all were found to interact specifically with alpha 2. These results suggest a model for TPR-protein interactions and for the role that a tandem array of TPRs may have in mediating transcriptional repression. PMID- 7498788 TI - Combinatorial signaling by Sonic hedgehog and Wnt family members induces myogenic bHLH gene expression in the somite. AB - We have demonstrated previously that a combination of signals from the neural tube and the floor plate/notochord complex synergistically induce the expression of myogenic bHLH genes and myogenic differentiation markers in unspecified somites. In this study we demonstrate that Sonic hedgehog (Shh), which is expressed in the floor plate/notochord, and a subset of Wnt family members (Wnt 1, Wnt-3, and Wnt-4), which are expressed in dorsal regions of the neural tube, mimic the muscle inducing activity of these tissues. In combination, Shh and either Wnt-1 or Wnt-3 are sufficient to induce myogenesis in somitic tissue in vitro. Therefore, we propose that myotome formation in vivo may be directed by the combinatorial activity of Shh secreted by ventral midline tissues (floor plate and notochord) and Wnt ligands secreted by the dorsal neural tube. PMID- 7498790 TI - The Drosophila abrupt gene encodes a BTB-zinc finger regulatory protein that controls the specificity of neuromuscular connections. AB - Motor axons make synaptic connections with specific muscles, and this specificity unfolds during development as motoneuron growth cones make specific pathway choices and ultimately recognize and synapse on their specific muscle targets. The Drosophila clueless mutation was identified previously in a genetic screen for mutations that disrupt motoneuron guidance and connectivity. We show here that clueless is allelic to abrupt. The abrupt gene is required for the embryonic formation of specific synaptic connections between a subset of motoneurons and a subset of muscles. Mutations in abrupt also reveal its role in establishing and maintaining muscle attachments, adult sensory cell formation, and morphogenesis of adult appendages. The abrupt gene encodes a zinc finger protein with a conserved BTB domain. Abrupt is expressed in muscle nuclei but not motoneurons, suggesting that abrupt controls the muscle expression of molecules required for correct motoneuron targeting, as well as molecules required for correct muscle attachments. PMID- 7498789 TI - Disruption of BMP signals in embryonic Xenopus ectoderm leads to direct neural induction. AB - Bone morphogenetic proteins (BMPs), which have been implicated in the patterning of mesoderm, are members of the transforming growth factor-beta (TGF-beta) superfamily. We have investigated the roles of Xenopus BMP-7 (XBMP-7) and BMP-4 (XBMP-4), and activin (another TGF-beta-related molecule) in early development by generating dominant-negative versions of these growth factors. Mutations were generated by altering the cleavage sites that are required for maturation of the active dimeric forms of XBMP-7, XBMP-4, and activin. These mutant constructs, designated Cm-XBMP-7, Cm-XBMP-4, and Cm-activin, result in polypeptides that allow for dimerization of the subunits, but are incapable of maturation. Expression of Cm-XBMP-7 and Cm-XBMP-4, but not Cm-activin, in the ventral marginal zone of the Xenopus embryo results in the development of a secondary axis, similar to that seen by ectopic expression of the truncated BMP receptor. These results suggest that the cleavage mutants interfere with BMP signaling during mesodermal patterning. We also found that expression of Cm-XBMP-7 or Cm XBMP-4 in animal cap ectoderm directly induces neuroectoderm. The neural induction was specific for Cm-XBMP-7 and Cm-XBMP-4 because ectopic expression of Cm-activin or Vg-1 did not mimic the same phenotype. Molecular study of neural patterning by Cm-XBMP-7 and Cm-XBMP-4 revealed that only anterior neuroectodermal markers are expressed in response to these Cm-XBMPs. These results suggest that the BMPs are involved in the specification of ectoderm in Xenopus development, and that neural induction requires the removal of BMP signals in the ectoderm. We propose that neural induction occurs by a default mechanism, whereby the inhibition of BMP signaling is required for the conversion of ectoderm to neuroectoderm in the developing Xenopus embryo. PMID- 7498791 TI - Mutation of RGA1, which encodes a putative GTPase-activating protein for the polarity-establishment protein Cdc42p, activates the pheromone-response pathway in the yeast Saccharomyces cerevisiae. AB - We have selected yeast mutants that exhibit a constitutively active pheromone response pathway in the absence of the beta subunit of the trimeric G protein. Genetic analysis of one such mutant revealed that it contained recessive mutations in two distinct genes, both of which contributed to the constitutive phenotype. One mutation identifies the RGA1 locus (Rho GTPase activating protein), which encodes a protein with homology to GAP domains and to LIM domains. Deletion of RGA1 is sufficient to activate the pathway in strains lacking the G beta subunit. Moreover, in wild-type strains, deletion of RGA1 increases signaling in the pheromone pathway, whereas over-expression of RGA1 dampens signaling, demonstrating that Rga1p functions as a negative regulator of the pheromone response pathway. The second mutation present in the original mutant proved to be an allele of a known gene, PBS2, which encodes a putative protein kinase that functions in the high osmolarity stress pathway. The pbs2 mutation enhanced the rga1 mutant phenotype, but by itself did not activate the pheromone pathway. Genetic and two-hybrid analyses indicate that an important target of Rga1p is Cdc42p, a p21 GTPase required for polarity establishment and bud emergence. This finding coupled with recent experiments with mammalian and yeast cells indicating that Cdc42p can interact with and activate Ste20p, a protein kinase that operates in the pheromone pathway, leads us to suggest that Rga1p controls the activity of Cdc42p, which in turn controls the magnitude of signaling in the pheromone pathway via Ste20p. PMID- 7498792 TI - A tactile sensory system of Myxococcus xanthus involves an extracellular NAD(P)(+)-containing protein. AB - CsgA is a cell surface protein that plays an essential role in tactile responses during Myxococcus xanthus fruiting body formation by producing the morphogenic C signal. The primary amino acid sequence of CsgA exhibits homology with members of the short-chain alcohol dehydrogenase (SCAD) family and several lines of evidence suggest that NAD(P)+ binding is essential for biological activity. First, the predicted CsgA secondary structure based on the 3 alpha/20 beta-hydroxysteroid dehydrogenase crystal structure suggests that the amino-terminal portion of the protein contains an NAD(P)+ binding pocket. Second, strains with csgA alleles encoding amino acid substitutions T6A and R10A in the NAD(P)+ binding pocket failed to develop. Third, exogenous MalE-CsgA rescues csgA development, whereas MalE-CsgA with the amino acid substitution CsgA T6A does not. Finally, csgA spore yield increased approximately 20% when containing 100 nM of MalE-CsgA was supplemented with 10 microM of NAD+ or NADP+. Conversely, 10 microM of NADH or NADPH delayed development for approximately 24 hr and depressed spore levels approximately 10%. Together, these results argue that NAD(P)+ binding is critical for C-signaling. S135 and K155 are conserved amino acids in the catalytic domain of SCAD members. Strains with csgA alleles encoding the amino acid substitutions S135T or K155R failed to develop. Furthermore, a MalE-CsgA protein containing CsgA S135T was not able to restore development to csgA cells. In conclusion, amino acids conserved in the coenzyme binding pocket and catalytic site are essential for C-signaling. PMID- 7498793 TI - The symmetry of the yeast U6 RNA gene's TATA box and the orientation of the TATA binding protein in yeast TFIIIB. AB - The central RNA polymerase III (Pol III) transcription factor TFIIIB is composed of the TATA-binding protein (TBP), Brf, a protein related to TFIIB, and the product of the newly cloned TFC5 gene. TFIIIB assembles autonomously on the upstream promoter of the yeast U6 snRNA (SNR6) gene in vitro, through the interaction of its TBP subunit with a consensus TATA box located at base pair 30. As both the DNA-binding domain of TBP and the U6 TATA box are nearly twofold symmetrical, we have examined how the binding polarity of TFIIIB is determined. We find that TFIIIB can bind to the U6 promoter in both directions, that TBP is unable to discern the natural polarity of the TATA element and that, as a consequence, the U6 TATA box is functionally symmetrical. A modest preference for TFIIIB binding in the natural direction of the U6 promoter is instead dictated by flanking DNA. Because the assembly of TFIIIB on the yeast U6 gene in vivo occurs via a TFIIIC-dependent mechanism, we investigated the influence of TFIIIC on the binding polarity of TFIIIB. TFIIIC places TFIIIB on the promoter in one direction only; thus, it is TFIIIC that primarily specifies the direction of transcription. Experiments using TFIIIB reconstituted with the altered DNA specificity mutant TBPm3 demonstrate that in the TFIIIB-U6 promoter complex, the carboxy-terminal repeat of TBP contacts the upstream half of the TATA box. This orientation of yeast TBP in Pol III promoter-bound TFIIIB is the same as in Pol II promoter bound TFIID and in TBP-DNA complexes that have been analyzed by X-ray crystallography. PMID- 7498794 TI - Genetic strategy for analyzing specificity of dimer formation: Escherichia coli cyclic AMP receptor protein mutant altered in its dimerization specificity. AB - Many transcriptional regulators function in homo- or heterodimeric combinations. The same protein can carry out distinct regulatory functions depending on the partner with which it associates. Here, we describe a mutant of the Escherichia coli cAMP receptor protein (CRP) that has an altered dimerization specificity; that is, mutant/mutant homodimers form preferentially over wild-type/mutant heterodimers. CRP dimerization involves the formation of a parallel coiled-coil structure, and our CRP mutant bears an amino acid substitution affecting the first "d" position residue within the alpha-helix that mediates CRP dimerization. The genetic strategy we used to isolate this CRP altered dimerization specificity (ADS) mutant is generalizable and could be utilized to isolate ADS mutants of other dimeric transcriptional regulators. PMID- 7498795 TI - A common function for mRNA 5' and 3' ends in translation initiation in yeast. AB - The mRNA poly(A) tail and its associated poly(A) binding protein (Pab1p) are ubiquitous in eukaryotes. The function of the poly(A) tail is to stabilize mRNA and to stimulate its translation. The development of a poly(A)- and cap-dependent yeast in vitro translation system has allowed us to understand how poly(A) stimulates translation. We find that Pab1p but not the cap binding protein eIF-4E is required for poly(A) tail-dependent translation, and that the Pab1p-poly(A) tail complex functions to recruit the 40S ribosomal subunit to the mRNA. These data introduce a new step into the pathway of translation initiation and merge the translational functions of the two ends of mRNA. PMID- 7498796 TI - Poly(A) site selection in the HIV-1 provirus: inhibition of promoter-proximal polyadenylation by the downstream major splice donor site. AB - In common with all retroviruses, the human immunodeficiency virus type 1 (HIV-1) contains duplicated long terminal repeat (LTR) sequences flanking the proviral genome. These LTRs contain identical poly(A) signals, which are both transcribed into RNA. Therefore, to allow efficient viral expression, a mechanism must exist to either restrict promoter-proximal poly(A) site use or enhance the activity of the promoter-distal poly(A) site. We have examined the use of both poly(A) sites using proviral clones. Mutation of the previously defined upstream activatory sequences of the 3' LTR poly(A) site decreases the efficiency of polyadenylation when placed in competition with an efficient downstream processing signal. However, in the absence of competition, these mutations have no effect on HIV-1 polyadenylation. In addition, the 5' LTR poly(A) site is inactive, whereas a heterologous poly(A) site positioned in its place is utilized efficiently. Furthermore, transcription initiating from the 3' LTR promoter utilizes the 3' LTR poly(A) signal efficiently. Therefore, the main determinant of the differential poly(A) site use appears to be neither proximity to a promoter element in the 5' LTR nor the presence of upstream activating sequences at the 3' LTR. Instead, we show that the major splice donor site that is immediately downstream of the 5' LTR inhibits cleavage and polyadenylation at the promoter proximal site. The fact that this poly(A) site is active in a proviral clone when the major splice donor site is mutated suggests that the selective use of poly(A) signals in HIV-1 is mediated by a direct inhibition of the HIV-1 poly(A) site by downstream splicing events or factors involved in splicing. PMID- 7498797 TI - A near-miss in Congress. PMID- 7498798 TI - Pigmented lesion with a raised center. PMID- 7498799 TI - Late-onset asthma: making the diagnosis, choosing drug therapy. AB - Physicians often fail to diagnose asthma in their older patients and instead attribute respiratory symptoms to aging alone or to other common diseases of the elderly. Although asthma is a common respiratory disease in older patients, its pathophysiology is poorly understood. Most elderly asthmatics are former smokers and have evidence of allergic disease. Asthma itself does not appear to increase mortality risk for the older patient. Spirometric results may show mild reversible airflow obstruction or fixed obstruction. The recommended therapy in the older patient follows that used for younger asthmatics. Inhaled corticosteroids are first-line agents, followed by inhaled anticholinergics. Smoking cessation, proper nutrition, and exercise are important nondrug therapies. PMID- 7498800 TI - Nursing homes: what you need to know for quality patient care. AB - The United States has 20,000 nursing homes and 1.5 to 2 million nursing home beds. Your choice of a nursing home often depends on its distance from the office and the preferences of colleagues and patients. The quality of care in a particular facility may be suggested by the bacteria found in urine, blood, and skin cultures from symptomatic patients. Communication with the nursing home staff is key to a well-organized practice. For quality of care and efficient reimbursement, limit the number of patients seen per visit. Federal law controls the use of physical restraints and drugs to manage problem behaviors. PMID- 7498801 TI - Low vision: physician screening helps to improve patient function. AB - Low vision is a common problem of older patients. As a primary care practitioner, you are not expected to treat most ocular problems, but you can play an important role by identifying patients with visual impairment. Familiarize yourself with the normal visual changes that occur with aging as well as the more common age related ocular diseases, including glaucoma, cataracts, age-related macular degeneration, and diabetic retinopathy. You can identify patients with visual problems with a careful history and basic in-office tests of visual acuity. Refer patients found to have visual impairment to an eye care specialist for further evaluation. PMID- 7498803 TI - [Indications for an Expert Team from the Polish Diabetes Society for early diagnosis of gestational diabetes mellitus]. PMID- 7498802 TI - [Diabetes mellitus in conceptional age and gestational diabetes mellitus in the St. Vincent Declaration]. PMID- 7498804 TI - [A model for medical care of gestational diabetes]. AB - Our approach to taking care of 122 diabetic pregnant women was presented. At first, they were hospitalized in the Pathological Pregnancy Ward. An intensive insulin therapy (human insulin) was initiated and all the patients were educated. The pregnant appeared as an outpatient every fortnight within the first part of pregnancy and every seven to ten days in the second one. They were hospitalized for a few days in 12th and 24th week of pregnancy and from 36th/37th week until delivery. Unfortunately, their first arrival to our center was too late, usually with diabetes out of control. There were 3 miscarriages (2.4%), 111 newborns were in a satisfactory condition (91%), 4 died (3.3%), congenital malformations were found in four others (3.3%). A successful course of pregnancy was dependent upon a proper care taken of patients early enough, good cooperation with them and related with it satisfactory metabolic control of diabetes. PMID- 7498806 TI - [Acid-base balance in capillary blood of newborns with perinatal hypoxemia symptoms]. AB - The purpose of the study was to compare acid-base parameters in capillary blood of newborns from groups with hypoxaemia symptoms and from the control group (physiological deliveries). 111 newborns were divided into 8 groups. Group I consisted of 33 newborns from uncomplicated labour while groups II-VIII involved newborns from complicated labor with symptoms of hypoxaemia. Capillary blood was taken in the 10th and 60th minute and after 24 hours of newborn's life. In newborns with pathological CTG, green amniotic fluid and prolonged first stage of labour--higher duration of acidosis was observed. PMID- 7498805 TI - [Changes in levels of human placenta lactogen (hPL), progesterone, and estriol in blood serum and estrogens in urine during gestational diabetes mellitus]. AB - The aim of the study was estimation of endocrinological function of placenta in pregnancy complicated by GDM. The study were performed on a group 13 women with GDM and 14 women in normal pregnancy. All women with GDM were treat by diet and intensive insulinotherapy with self monitoring levels of glucose. In women with GDM level of fructosamine and HbAlc were significant higher but in normal range. In 28, 36, 37, 38, 39 week of pregnancy were determined levels of hPL, oestriol and progesterone in serum and daily excretion of oestrogens in urine. From 36 week of pregnancy levels of hPL, oestriol and progesterone were significant lower in women with GDM. In 39 week of pregnancy level of hPL was 9.34 micrograms/ml vs. 12.71 micrograms/ml, p < 0.01, oestriol was 495.77 nmol/l vs. 681.14 nmol/l, p < 0.01 and progesterone was 25.12 ng/ml vs. 34.52 ng/ml, p < 0.01 respectively in women with GDM and in normal pregnancy. PMID- 7498808 TI - [Modification of operative support methods for urinary stress incontinence in women using a free fascia band]. AB - The author modified the sustaining methods applied until now for treatment of stress urinary incontinence (s.u.i.) in women by replacing the peduncular "supports" with a free fascial band. The surgeries by choice were performed on hard working manual female workers in which s.u.i. was associated with anterior colpoptosis. During 6-years observation, recovery (very good and good results) was found in 89.3% of cases. PMID- 7498807 TI - [Estimation of health status in infants fed naturally within the Zoz Koscian region]. AB - Lately the problem of breast-feeding of babies has been coming into the domain of great interest. Basing on the inquiry, methods of breast-feeding of babies at the age up to one and their influence on morbidity rate caused by infection diseases has been investigated. A population of babies at the age up to one living in Koscian city and surrounded villages has been tested. All the children were born with no complications within delivery and infant period. All data were elaborated using statistical methods. The results are tabulated and shown in charts. The results proved that breast-feeding babies are subject to less amount of diseases compared with babies fed by artificial food. Much more frequently a hospital service was needed for babies fed in artificial mode. Summing up in general it can be clearly stated that breast-feeding reinforces babies against all sorts of infections. In that connection breast-feeding mode should be widespread by all pediatrics. PMID- 7498812 TI - [Changes in brain tissue resulting from the HELLP syndrome verified by CT examination]. AB - New noninvasive imaging methods are very effective in the detection and differentiation of anatomic characteristics and accompanying symptoms observed in the course of the Hellp syndrome. Within the clinical picture presented the evidence of brain injury as a result of intravascular coagulopathy was predominant. CT confirmed the presence of brain lesions. PMID- 7498811 TI - [Does use of mammography and fine-needle biopsy help in early detection of breast carcinoma?]. AB - Two diagnostic methods: mammography and fine-needle biopsy were used as an early detecting method in 482 cases of breast carcinoma. The outcome of examinations showed high diagnostic and prophylactic values. PMID- 7498809 TI - [Radical vulvectomy using Way's method with bilateral removal of inguinal, thigh, iliac and veil lymph nodes in the clinic]. AB - Thirty-one cards of women after radical vulvectomy in Way's mode were analysed. Metastases in inguinal, thigh, iliac and veil lymph nodes were found only in 19.35%. Authors conclude that removal of iliac and veil lymph nodes can be given up after previously negative outcome of intra-operative examination of inguinal and thigh lymph nodes. Mean period of patient's staying in hospital was long- about 40 days. In 51.6% of cases the blood group was "A", with the mean for the whole country 40.5%. PMID- 7498810 TI - [Treatment of condylomata acuminata of the male reproductive organs with CO2 laser under colposcopic control]. AB - The first results of laser therapy in treatment of condylomata acuminata of male reproductive organs are reported. Between November 1991 and December 1993 in Department of Reproduction, Institute of Gynecology and Obstetrics, Karol Marcinkowski University School of Medical Sciences, Poznan in nine patients with condylomatous changes of reproductive organs CO2-laser therapy under colposcopic control was done. In 7 of them the healing was achieved. In two patients second laser therapy was done. All patients are still under control in Department of Reproduction and there was no recurrence diagnosed. PMID- 7498814 TI - Human needs, health care, and nursing practice. PMID- 7498813 TI - [A case of epithelioid sarcoma of Bartholin's glands]. AB - A case of vulvar epithelioid sarcoma in 49-year old female was described. The disease originated from Bartholin's gland. The patient was treated by radical surgery and then multidrug chemotherapy and radiotherapy. In spite of intensive, multimodality treatment, the patient died after 8 months. PMID- 7498815 TI - Diagnosis: deficit in independent living. PMID- 7498816 TI - Posttraumatic stress disorder and the World War II veteran. AB - Throughout history, soldiers have suffered the psychologic consequences of war. PTSD in the WWII combat veteran presents with physical, psychologic, and social manifestations. Specific physical problems, such as traumatic arthritis and digestive disorders, may be linked to POW experiences. Psychologic symptoms include denial, numbing, intrusive images and thoughts, and nightmares. The older combat veteran may experience long-delayed PTSD symptoms when facing the losses associated with aging, loss of loved ones and declining physical health. Nurses are challenged to discover which elderly patients are combat veterans, what their military experiences were, how the experiences are affecting their mental and physical health, and whether biopsychosocial factors are concurrently affecting the health of the patients known to have PTSD. PMID- 7498817 TI - Benefits of dehydration in terminally ill patients. PMID- 7498818 TI - Consequences of visual impairment on leisure activities of the elderly. PMID- 7498819 TI - Pacemaker assessment and care plans in long-term care. AB - Residents with implanted pacemakers are a real challenge, especially to nurses, in long-term care. The facility is accountable for the quality and timeliness of care and service to the resident. Documenting the physical assessment, developing and implementing a significant plan of care, and monitoring that care, for a resident with a pacemaker, are serious professional nursing responsibilities. PMID- 7498820 TI - Elders of America ... who will care? PMID- 7498821 TI - Jumping the hurdles of high-tech home care. PMID- 7498823 TI - Interventions for osteoporosis. PMID- 7498822 TI - Eating out. PMID- 7498824 TI - [Maternal serum screening for detection of fetal chromosome abnormalities]. PMID- 7498825 TI - [Noninvasive prenatal diagnosis of fetal cells in maternal blood]. PMID- 7498827 TI - [Potential 3-dimensional (3D-)ultrasound in the diagnosis of fetal abnormalities]. PMID- 7498826 TI - [Reliability of invasive prenatal diagnosis in the first trimester in comparison with standard amniocentesis]. PMID- 7498828 TI - [Applications of magnetic resonance tomography (MRT) in obstetrics]. PMID- 7498829 TI - [Intrauterine diagnosis and therapy of fetal arrhythmias]. PMID- 7498830 TI - [Diagnostic-therapeutic concept in hydrocephalus]. PMID- 7498831 TI - [Diagnostic-therapeutic concept in abnormalities of the kidneys and efferent urinary tract]. PMID- 7498832 TI - [Diagnostic-therapeutic concept in abnormalities of the abdominal wall and diaphragm]. PMID- 7498833 TI - [Observations on ethics in prenatal diagnosis from the viewpoint of the pediatrician]. PMID- 7498834 TI - [Undetected tubal pregnancy in gynecologic practice. A case from expert witness files]. PMID- 7498835 TI - [26th Annual meeting of the German Society of Plastic Surgery. Leipzig, 20-23 September 1995. Abstracts]. PMID- 7498836 TI - [Branchiogenic anomalies (with special reference to severe and complicated outcome)]. AB - Lateral cervical fistulae are usually described as examples for inborn branchiogenic anomalies. They result from evolutionary disorders of the first and second branchiogenic arch. Four patients were seen with cervical fistulae, three of those having a recurrence after surgical treatment. After a short review of the evolution and the topographic anatomy of such cysts and ducts, four patients are presented with all the details regarding their partially very complicated history. PMID- 7498837 TI - [Long-term outcome of the donor site after free, microvascular pedicled iliac crest transplantation: experiences with 95 cases]. AB - Using the anterior iliac crest as donor-site for free osseous, osseomuscular, and osseomyocutaneous flaps has proven itself an important reconstructive instrument both in maxillofacial and in orthopaedic surgery. We present a long-term retrospective evaluation of the donor-site situation after vascularized free anterior iliac crest transfer. The study is based in 95 cases from the years 1985 to 1993 including 11 cases from Switzerland (Kantonsspital Aarau). The patients were examined, looking for objective complications and asked questions about their subjective symptoms at the donor site. These objective and subjective results were then correlated. Herniation developed in 3% of cases, loss of sensibility in the region of the N. cutaneous femoris lateralis occurred in 9.5% and was in 6% reversible. Scars were in 7% aesthetically not satisfying with no keloids or hypertrophic scarring after all wounds healed per primam intentionem. Mostly minor alteration of the outer surface of the donor site was notable in 20 to 25% of the patients. Pain was a problem for an average of 7% of cases. Impairment in private life was named in 3.5%, in professional life in 4.7%, and in daily life in 11.9% of cases. Not content with the donor-site situation were 13.5%, not content with the preoperative information were 12% of the patients. Nonetheless were 98.8% of the patients willing to, if necessary, undergo the same operation again. Surprisingly, there is a negative correlation between objective and subjective results: Merely 25% of the patients with sensibility loss were not content with the donor-site result of the operation and vice versa only 1% of the dissatisfied patients had objectively measurable minor complications. PMID- 7498838 TI - [Lyme borreliosis in hand surgery. A case report]. AB - The clinical courses of Borrelia burgdorferi infections may show such variety, that diagnostic problems arise. On the other hand, early antibiotic therapy is necessary in order to prevent late-term complications. In this article, we present an exceptional case of a long-term documentation of Lyme-disease which was not diagnosed until an acrodermatitis chronica atrophicans had developed. Whether the manifestation of bilateral carpal tunnel syndrome is connected with Lyme borreliosis is yet to be clarified. PMID- 7498839 TI - [Fracture of the lunate--a rare injury]. AB - Fractures of the normal lunate--isolated or associated with fractures of the scaphoid or radius--are very rare. In the Traumatological Hospital Meidling/Vienna, 12 patients with acute fractures of the lunate bone were treated between 1983 and 1993. Dependent on the fracture-line and the intraosseous vascularity, partial or total avascular necrosis of the bone occurs in rare cases. In dislocated pole fractures, surgical intervention is indicated. PMID- 7498840 TI - [Fracture of the lunatotriquetrum bone in the triquetral part. Case report and review of the literature]. AB - The fusion of the lunate and triquetrum is the most common type of carpal fusion. Fractures of the os lunatotriquetrum are rare and usually occur at the point of union. In our case we found an isolated transverse fracture of the triquetral part of the fused bone without dislocation. In case of non-union, the differential diagnosis of an incomplete fusion must be taken into consideration. PMID- 7498841 TI - [Intermediate-term treatment outcome after fractures of the proximal radius]. AB - The treatment of fractures of the proximal radius is controversial. This retrospective elaborates on indications for operative or conservative treatment, dependent on fracture type. A new classification of fracture and a new evaluation score for follow-up examination are presented. The study shows very good results for conservative treatment of type A1-fractures without dislocation of fragments and good results for operative treatment of type A2-fractures. Type A3-fractures (multifragment fractures) demand reconstruction. If that is not possible, the primary resection of the radial head leads to better results than its secondary resection. The implantation of silastic prostheses of the radial head leads to worse results in primary (2 cases) and secondary resection (2 cases) than resection of radial head alone. PMID- 7498842 TI - [External fixation for support in soft tissue reconstruction in hand surgery]. AB - A mini fixator system was used to assist the operative correction of soft-tissue flexion contractures. The external fixator served as a tool for gradual correction and to secure joint position after soft-tissue release. Following wound healing, certain fixator components were unlocked and joint mobilisation initiated. Correction involved a contraction of the first web-space as well as flexion contractures of the index, middle, ring, and small fingers. The fixators were removed after four to six weeks. Complications related to the fixator equipment were not observed. The spread from thumb to index finger tip was increased from 3.5 to 16.5 cm. The functional result in sustained grip strength reached a value of 82% compared to the uninjured opposite side. PMID- 7498843 TI - [Preoperative continuous extension treatment of very severe Dupuytren's contractures]. AB - The continuous extension of Dupuytren's contracture prior to aponeurectomy in severe cases, introduced by Messina, is presented. This technique demonstrates the possibility of reversing contractures; it simplifies the surgical treatment thereafter and avoids difficulties in blood circulation and sensibility. First results in ten patients are presented. PMID- 7498844 TI - [Description of Dupuytren disease by the Basel physician and anatomist Felix Plater in 1614]. AB - Plater's description of Dupuytren's disease in 1614 is explained with regard to his understanding of the anatomy. The current view that Plater believed the disease to be caused by dislocation and shortening of the flexor tendons is based upon misinterpretation of the original Latin text. With the help of his anatomical studies, Plater had proven that subcutaneous ligamentous extensions of the palmar aponeurosis and not the flexor tendons were responsible for Dupuytren's disease. Felix Plater realised more than one hundred and fifty years before Cline, Cooper, and Dupuytren the palmar aponeurosis as the anatomical substrate of the disease. PMID- 7498845 TI - [Updated status of replantation services in Germany, Austria and Switzerland]. AB - The revised survey involves the years 1991 and 1992 and includes 41 university and major civil hospitals. As in the years 1989 and 1990, replantation activity in the year 1991 still declined, whereas in 1992 we registered a sharp increase in replantations, especially in Germany. In numbers, 856 were reported for 1991 (72 macro- and 784 microreplantations), and 1091 in 1992 (85 macro- and 1006 microreplantations). A considerable number of amputation cases (617) were rejected mainly due to lack of operative manpower capacity. PMID- 7498846 TI - [Dysmorphophobia--a disorder of modern times?]. PMID- 7498847 TI - [Parvovirus B19--a ubiquitous, little know virus. From erythema infectiosum and arthropathy to chronic anemia]. PMID- 7498848 TI - [St. John's wort as antidepressive therapy]. AB - St. John's Wort (Hypericum perforatum) has been used to treat a variety of complaints since ancient times. Recent studies have shown that it is clinically effective for the treatment of the symptoms of depression. It has proved superior to placebo, equally as effective as standard medication and has a clear advantage over the latter in terms of side-effects. It follows that, on the basis of our present knowledge, St. John's Wort can be recommended for use as an anti depressant. PMID- 7498849 TI - [Dermatomycoses. 7: Mycoses of the trunk, neck and face--clinical picture and differential diagnosis]. PMID- 7498850 TI - [Therapy of candiduria by alkalinization of urine. Oral treatment with potassium sodium-hydrogen citrate]. AB - METHOD: Eighteen hospitalized patients with candiduria were treated with oral potassium-sodium-hydrogen citrate to alkalinize the urine. The results obtained were compared with those observed in an untreated retrospective control group. Dosage was adjusted in accordance with the pH of the urine measured immediately before treatment with the aim of achieving a pH of 7 to 7.5. RESULTS: All patients had an indwelling catheter, which is a predisposing factor for candiduria. In 16 out of 18 patients (89%) treatment with potassium-sodium hydrogen citrate raised pH and resulted in the disappearance of candiduria. Duration of treatment varied between two days and one month (mean: seven days). In four patients the urine became completely sterile; during treatment 12 out of 18 patients developed significant bacteriuria (in eight cases of these the indwelling catheter had been left in place). CONCLUSIONS: Alkalinization of the urine is a simple and effective method of treating candiduria in patients with an indwelling catheter. An additional advantage is the metaphylaxis and prophylaxis of renal stone formation in immobilized patients. PMID- 7498851 TI - [Basic therapy of osteoporosis in the elderly. Interview by Dr. rer. nat. Till Uwe Keil]. PMID- 7498852 TI - [Postmenopausal estrogens also for the heart! Postmenopausal hormone replacement- mechanisms of action]. PMID- 7498853 TI - [Hepatitis C virus infections in hospital roommates?]. PMID- 7498854 TI - [Gluten intolerance--investigating an iceberg?]. PMID- 7498855 TI - [Prediction of type I diabetes by antibody screening]. AB - For the prediction of type I diabetes in first degree relatives of type I diabetics, a variety of laboratory methods are now available. Thus, screening for serum antibodies against the body's own insulin, islet cells and the enzyme glutamate decarboxylase, represents a simple and well-established routine examination used for early diagnosis. The combination of various parameters such as e.g. the age of the person at risk, antibody status (nature, number, combination or titer of the antibodies) and capacity for early insulin secretion following i.v. glucose loading, permits us to predict, with quite good accuracy, the risk years before there are any clinical manifestations. These successes of early diagnosis have led to new concepts of immunoprophylaxis for the prevention of type I diabetes. Thus, at the present time two highly promising studies involving the prophylactic administration of insulin or nicotinamide in persons at risk are underway in Germany, the results of which to date suggest that a new epoch of immunotherapeutic treatment of this disease may be in the offering. PMID- 7498856 TI - [Neurolinguistic programming in physician-patient communication. Basic principles of the procedure--examples for application in surgery]. AB - Neurolinguistic programming (NLP) is a means of improving physician-patient communication that can be learned by any doctor. The present article first describes some of the fundamentals of NLP and then provides examples taken from the field of surgery-in the first instance dealing with the treatment of painful conditions by means of trance or dissociation and, secondly, on the influencing of expectations and the restructuring (reframing) of doctrines in a patient with malignant disease. PMID- 7498857 TI - [Dermatomycoses. 8: Mycoses of the trunk, neck and face--therapy and follow-up]. PMID- 7498859 TI - [Optimal medical performance develops on the decision tree. Decision methodology and analysis--clinical economy series, 6]. PMID- 7498858 TI - [Effectiveness and tolerance of combined verapamil retard and hydrochlorothiazide. Results of a double-blind, randomized study]. AB - AIM: To determine, in a multicenter study, whether the fixed combination of verapamil retard (240 mg) and hydrochlorothiazide (12.5 mg) is capable of lowering blood pressure more effectively than either of the substances alone or placebo, while retaining good tolerability. METHOD: A total of 173 patients who, after a four-week run-in period had a diastolic blood pressure of at least 100 and not more than 115 mm Hg, were randomized to six weeks of monotherapy with one of the two substances, placebo or the fixed combination. RESULTS: Combination treatment proved superior to monotherapy with either substance in lowering the diastolic blood pressure (p < 0.05). Also in terms of normalization (diastolic pressure < 90 mm Hg) the combination was clearly superior (p < 0.05). During the study no side effects beyond those associated with the single substances, occurred. PMID- 7498860 TI - [Endoscopy in ulcer: how often and why?]. PMID- 7498861 TI - [Effectiveness and costs of screening colonoscopy]. AB - Screening colonoscopy is always indicated when rectosigmoidoscopy reveals an adenoma, since this lesion roughly doubles the patient's risk of contracting colonic cancer. Follow-up should be performed at intervals of about three years after endoscopic removal of all colorectal polyps. Repeated screening examinations are recommended for the following genetic diseases that carry an increased risk of colorectal carcinoma: familial adenomatous polyposis (FAP) and its genetic variant, hereditary non-polyposis colorectal cancer syndrome (HNPCC) and hamartomatous polyposis syndromes (e.g. Peutz-Jeghers). Also in the case of familial "sporadic" carcinoma of the colon, regular screening colonoscopies for first degree relatives are recommended. Although the use of regular screening colonoscopies in patients with a long history of extensive ulcerative colitis is controversial, the recent results support such examinations. While the benefit of screening colonoscopy or sigmoidoscopy of the general population from the age of 50 onward must be affirmed, it should be weighed against the costs involved in such an undertaking. At the present time, the American Cancer Society recommends that from the age of 50 onward, the annual fecal test for occult blood should be supplemented by sigmoidoscopy performed every three to five years. PMID- 7498862 TI - [Dermatomycoses. 9: Mycoses in skin fold areas]. PMID- 7498863 TI - [Long-term treatment of chronic pain with tilidine-naloxone. An analysis of 50 patients with chronic pain conditions of non-malignant origin]. AB - METHOD: 50 patients with incurable chronic pain states were treated for a period of between six months and 14 years (median: 31 months) with tilidine-naloxone, and the results of tumor recorded. Amelioration of pain was achieved in an average of 60.7% of the cases. The 16 patients of this series suffering from neuropathic pain who received a somewhat lower dose responded equally as well (60.3% amelioration) as the overall group. In 24 patients suffering from pain, the daily dose remained unchanged throughout the course of treatment, had to be increased in 19, and was reduced in seven patients. In two patients, acceptable side-effects were indicated; in no case were there any signs of drug-induced organic damage. CONCLUSIONS: The results show that tilidine-naloxone is a highly effective opioid analgesic with a remarkably favorable benefit-risk ratio for use in long-term treatment. PMID- 7498864 TI - [Malignant melanoma of the skin]. AB - The incidence of malignant melanoma (MM) of the skin is on the increase. If the mortality of this disease is to be reduced, the identification of persons at risk, and early clinical diagnosis are of decisive importance. The prognosis depends on the stage of the disease and has improved over the last few decades. The present article provides an overview of the risk factors and discusses the latest guidelines for the diagnosis, treatment and follow-up, the aim of which is to make possible a uniform approach to the management of malignant melanoma. PMID- 7498865 TI - [Nitric oxide as agent with cardio-depressive effect]. PMID- 7498866 TI - [Dermatomycoses. 10: Mycoses of the mouth cavity and ear canals--clinical picture and differential diagnosis]. PMID- 7498867 TI - [Adrenal cortex insufficiency and hypothyroidism in advanced age]. AB - In geriatric patients symptoms of endocrine disorders easily can be regarded to be due to the patient's old age. This is demonstrated in a case of an 80-year-old woman suffering from the combination of adrenal insufficiency and hypothyroidism. Early recognition of typical clinical signs and adequate, simply performed replacement therapy results in a significant improvement of the patient's quality of life. PMID- 7498868 TI - [Treatment of depressive symptoms with a high concentration hypericum preparation. A multicenter placebo-controlled double-blind study]. AB - In a multicenter, placebo-controlled double-blind trial, the effect on depression (ICD 10 F 32.1) of treatment with an innovative highly concentrated hypericum preparation was investigated. The study contained 97 outpatients who received 100 to 120 mg of the hypericum extract bid. The course of the illness was assessed with the Hamilton Depression Scale (HAMD), the von Zerssen Depressivity Scale (D S) and the Clinical Global Impression Scale (CGIS). Treatment resulted in an appreciable improvement in the symptoms of depression, and the 70% response rate (n = 43), corresponded to that of chemical antidepressants. The preparation also showed an anxiolytic effect. The substance was extremely well tolerated, and no side-effects were reported by any of the patients. PMID- 7498869 TI - [Gene therapy for AIDS in an animal experiment]. PMID- 7498870 TI - [Behavior therapy--a current form of psychotherapy. Psychotherapy series, 10: Behavior therapy in primary care]. PMID- 7498871 TI - [Vascular surgery methods for elderly patients. Elective interventions with reduced risk are possible]. AB - As in other areas, the percentage of elderly patients consulting the vascular surgeon is also on the increase. In recent years, our diagnostic possibilities have been expanded by non-invasive procedures (e. g. colour-coded duplex sonography, NMR angiography) and by improvements in intravascular imaging procedures. Early diagnosis makes it possible to perform interventions electively, which is of particular importance for minimizing risks in the geriatric patient. In the case of supra-aortic interventions (e.g. carotid reconstruction), surgical treatment of an aneurysm, and in venous occlusions, the indication for surgery in the elderly should not be applied too restrictively. For the treatment of arterial occlusions, a fundamental choice must be made between interventional (e.g. PTA) and surgical procedures. When peripheral arterial occlusive disease presents, conservative therapy should be applied for as long as is possible, since over the long term this can save many patients the need for surgery. PMID- 7498872 TI - [Diagnostic imaging in senile dementia. Exclusion of specific treatable causes also in the elderly]. AB - Dementia is a common problem in elderly patients and may influence the indication for invasive diagnostic and therapeutic procedures under consideration for other diseases more than age itself. This short paper reviews the role of tomographic imaging techniques--whose possibilities have expanded in recent years--in the differential diagnosis of dementia. Computerized tomography (CT) of the brain is used primarily to detect such potentially treatable conditions as multiple ischemic infarctions, hematomas, hydrocephalus and brain tumors. It should be performed even in old age, unless the patient's general state of health is too poor to expect good results from therapeutic measures. Magnetic resonance imaging is more sensitive in ischemic white matter lesions and hippocampal atrophy, and should therefore be used preferentially in mildly affected patients. Functional imaging methods, such as SPECT and PET, are needed only to demonstrate impairment of cortical blood flow and metabolism in clinically unclear cases. PMID- 7498873 TI - [Kidney diseases in the elderly from the internal medicine viewpoint. Diagnosis and therapy]. AB - In the conservative and dialysis treatment of renal disease in the elderly, the methods employed are the same as those applied in the young patient. The various procedures for the qualitative and quantitative determination of protein in the urine are non-stressful and therefore of special value in the old patient. The diagnostic usefulness, in particular for follow-up observation of renal disease, is very high. With respect to the use of dialysis in the elderly, the various methods of peritoneal dialysis-CAPD, IPD and CCPD-have all proved beneficial. Since they do not particularly stress the circulatory system, they can be used in patients with coronary heart disease, arterial occlusive disease, or hypertension. For the choice of therapeutic procedure, the individual assessment of his life situation by the patient himself is of major importance. PMID- 7498874 TI - [Dermatomycoses. 11: Mycoses of the mouth cavity and ear canals--epidemiology and therapy]. PMID- 7498875 TI - State of the art and perspectives in hypertension. PMID- 7498876 TI - [Epidemiologic and medical sociological aspects of hypertension]. AB - Cardiovascular diseases are a major determinant of overall and premature mortality in advanced and in rapidly developing societies. In view of their importance it is mandatory to identify underlying risk factors and to guide preventive and therapeutic actions accordingly. Hypertension is one of the most prevalent and well-established cardiovascular risk factors. This paper briefly summarizes some major determinants of high blood pressure from an epidemiologic and sociomedical point of view. Determinants include age, overweight, physical inactivity, salt and alcohol intake, family history of hypertension, race and socio-economic status. The complexity of these influences is discussed by pointing to interactions between genetic and socio-environmental influences, e.g. in the case of age, bodyweight and salt intake. Based on experimental animal research there is now solid evidence on direct links between psychosocial stress, patterns of neuroendocrine activation and elevation of blood pressure. Four theoretical concepts are described which identify conditions of chronic psychosocial vulnerability or protection in man, and their role in explaining the prevalence of hypertension in epidemiologic studies is discussed. The four concepts are labelled "socioemotional support", "lifestyle incongruity", "job strain" and "effort-reward-imbalance at work". Special emphasis is given to the two latter concepts in view of the potential role of occupational life in triggering high blood pressure during middle adulthood. For instance, Table 1 indicates that high job strain, i.e. high demands in combination with low control at work, is associated with a relative risk of 3 of being hypertensive, after adjusting for important confounders.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7498877 TI - Genetics in arterial hypertension--clinical and experimental aspects. AB - Genetic approaches provide a new insight into the understanding of biological mechanisms underlying blood pressure regulation and hypertension. Some progress has been made in elucidating the molecular mechanisms of rare forms of hypertension. Common forms of hypertension are likely to be due to combinations of DNA variants of several genes involved in blood pressure regulation. Only little progress has been made in the identification of the underlying genetic factors, due to the complex nature of this disease. Experimental crosses of hypertensive rats offer an ideal setting for the genetic dissection of mammalian physiology. With the opportunity to study inbred rat strains the problem of genetic heterogeneity becomes irrelevant. Cosegregation studies investigating candidate genes for hypertension have been carried out for a number of genes. Moreover, systematic mapping of quantitative trait loci involved in blood pressure regulation has recently become possible in genetically hypertensive rat models. In contrast, genetic studies involving hypertensive individuals are currently limited to the investigation of candidate genes in association and sib pair analysis. So far, the angiotensinogen gene is the only locus that has been implicated in the pathogenesis of human hypertension in association as well as sib-pair analysis. Considering the complexities of genetic hypertension, animal models will play an important role in the genetic dissection of this disease. A critical prerequisite for the study of human hypertension is the availability of large patient and family cohorts. PMID- 7498878 TI - [Experimental types of hypertension--models for essential hypertension in the human?]. AB - Since the pathogenesis of essential hypertension is multifactorial, the causal therapy of primary arterial hypertension remains a great challenge. At a given genetic predisposition, the manifestation of hypertension depends critically on lifestyle factors. It is thus essential to study the molecular consequences of various deleterious lifestyle factors. We demonstrated by radiotelemetric measurements that an increased caloric intake raises both systolic and diastolic blood pressure as well as heart rate in spontaneously hypertensive rats (SHR). This model is comparable to hyperkinetic hypertension in hypertensive persons which, if it persists, will lead to established hypertension. Overfeeding also results in the characteristic metabolic derangements (hyperinsulinemia, hypertriglyceridemia) of insulin resistant hypertensive persons. The enhanced sympathetic outflow of the brain can be potentiated by lifestyle factors such as high sodium intake and psychological stress. In contrast to sodium intake, psychological stress (e.g. schedule-induced stress) is difficult to mimic in animal experiments. In view of the recent progress in the characterization of imidazoline receptors in the rostral ventrolateral medulla and the development of antihypertensive drugs with a high selectivity (moxonidine) for imidazoline receptors, efforts should be made to elucidate key regulatory mechanisms involved in brain insulin sensitivity and appetite regulation. Such an approach could help in pharmacologically reducing the influence of deleterious lifestyle factors at a given genetic predisposition. PMID- 7498879 TI - [Role of the cardiac renin-angiotensin system in hypertensive heart disease]. AB - Chronic mechanical stress of the heart by arterial hypertension is a primary cause of left ventricular hypertrophy. The cardiac renin-angiotensin system is often found activated in conditions of increased afterload or mechanical stress of myocytes. Cardiac expression of angiotensinogen and angiotensin-converting enzyme (ACE) is increased resulting in elevated cardiac angiotensin II formation. This has been demonstrated in stretched cardiac myocytes in vitro as well as in animal models of pressure overload hypertrophy (supravalvular aortic stenosis and spontaneously hypertensive rats) and in human pressure overload hypertrophy (aortic stenosis). Functional consequences of elevated angiotensin II levels may be vasoconstriction of the coronary vasculature and a deterioration of diastolic function of the hypertrophied heart. Local formation of angiotensin II may also have proliferative effects on cardiac myocytes and connective tissue cells. Angiotensin II may, thus, be an important factor causing development and progression of left ventricular hypertrophy by itself. Blockade of the renin angiotensin system has been found to be an effective treatment of hypertensive heart disease, probably better than any other antihypertensive medication. Regression of left ventricular hypertrophy has been achieved by blockade of the angiotensin II pathway either by administration of an angiotensin-converting enzyme inhibitor or an angiotensin II type 1-receptor blocker. The animal model of supravalvular aortic stenosis has been used to show beneficial effects of blockade of the renin-angiotensin system on hypertrophy and survival. In this rat model regression of left ventricular hypertrophy by ACE-inhibition or angiotensin II type 1-receptor blockade could be demonstrated by serial echocardiographic analyses while afterload of the left ventricle was still elevated.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7498880 TI - [Cardiac structure-function relationship and the renin-angiotensin-aldosterone system in hypertensive heart disease]. AB - Based on the epidemiologic data of the Framingham heart study, arterial hypertension and coronary artery disease are the most frequent etiologic factors for the development of heart failure. In the pressure overloaded heart, hypertrophic growth of the myocardium includes the enlargement of cardiac myocytes stimulated by ventricular loading. Non-myocyte cell growth involving cardiac fibroblasts may also occur but is not primarily regulated by the hemodynamic load. Cardiac fibroblast activation is responsible for the accumulation of fibrillar type I and type III collagens within the interstitium while vascular smooth muscle cell growth accounts for the medial thickening of resistance vessels. This remodeling of the cardiac interstitium represents a major determinant of pathological hypertrophy in that it accounts for abnormal myocardial stiffness and impaired coronary vasodilator reserve, leading to ventricular diastolic and systolic dysfunction and ultimately to the appearance of symptomatic heart failure. Several lines of evidence suggest that the renin angiotensin-aldosterone system is involved in regulating the structural remodeling of the nonmyocyte compartment, including the cardioprotective effects of angiotensin converting enzyme (ACE) inhibition that was found to prevent myocardial fibrosis in the rat with renovascular hypertension. In rats with genetic hypertension, established left ventricular hypertrophy, abnormal diastolic stiffness due to interstitial fibrosis, and reduced coronary vasodilator reserve associated with medial wall thickening of intramyocardial resistance vessels, the ACE inhibitor lisinopril was able to restore myocardial structure and function to normal. These cardioreparative properties of ACE inhibition may be valuable in reversing left ventricular dysfunction in hypertensive heart disease. PMID- 7498881 TI - [3-D echocardiography reconstruction of the thoracic aorta]. AB - Since the beginning of the nineties the combination of echocardiography equipment with a computer algorithm for 3-D reconstruction is commercially available. The initial experiences with this unit (TomTec-system, Munich, Germany) in clinical and intraoperative settings are reported. The described unit consisted of a specially constructed transesophageal echo probe, in which the transducer (5 MHz) is mounted on a sliding carriage allowing free up and down movement in axial directions. The transducer obtains digitally dynamic 3-D information by automated pull-back of the transducer in 0.67 mm slices, gated to ECG and respiration. The information is stored on a hard disk and in the computer RAM. The acquisition time--depending on different trigger intervals--ranges from 5 to 10 minutes, usually not longer than for conventional TEE. The images are stored in special 4 D (that is 3-D in motion) data sets, allowing the calculation of any desired plane in real-time at a triggered ECG phase or dynamically. Thus this technique gives the capability of retrospective reconstruction of any view into the aorta in contrast to the omniplane TEE examination, the so-called anyplane echocardiography. In the clinical management of patients with thoracic aortic diseases transesophageal echocardiography (TEE) has become the central diagnostic tool as well as for the follow-up. This is caused by its high sensitivity and specificity comparable with that obtained by MRI. Nevertheless in a small percentage of cases there remains a questionable situation concerning the morphology of the distal ascending aorta and the aortic arch illuminated by the use of 3-D reconstruction. PMID- 7498882 TI - [Assessment of myocardial vitality with FDG-PET: still research or ready for general practice?]. PMID- 7498884 TI - Can HMOs manage the mental health benefit? PMID- 7498883 TI - Estimating costs of mental health and substance abuse coverage. AB - The cost of expanding mental health and substance abuse treatment coverage is a major impediment to reforming insurance coverage for these types of conditions. The recent experience with national health care reform offers a case study in cost estimation for mental health and substance abuse coverage. The impact of managed care and the cost of expanding coverage to currently uninsured persons introduced uncertainty into predictions. This paper critically reviews that experience and draws lessons for estimating future costs of policy initiatives. PMID- 7498885 TI - What did the health care reform debate mean for mental health policy? PMID- 7498886 TI - Children and mental health: how can the system be improved? PMID- 7498887 TI - Ability to obtain health care: recent estimates from the Robert Wood Johnson Foundation National Access to Care Survey. AB - This DataWatch presents findings on Americans' ability to obtain health care. Data from the 1994 National Access to Care Survey sponsored by The Robert Wood Johnson Foundation suggest that earlier studies have underestimated the access problems facing Americans by not asking about supplementary services such as prescription drugs, eyeglasses, dental care, and mental health care or counseling. Using this more inclusive definition of health care needs, we estimate that 16.1 percent of Americans were unable to obtain at least one service they believed they needed. While income is highly correlated with unmet need, most persons reporting access problems are not poor. PMID- 7498888 TI - Children's mental health service use across service sectors. AB - This DataWatch explores the roles of human service sectors (mental health, education, health, child welfare, and juvenile justice) in providing mental health services for children. The data are from the first wave of the Great Smoky Mountains Study of Youth, a population-based study of psychopathology and mental health service use among children. The results show somewhat higher rates of mental health service use than has been reported previously, while continuing to show a substantial amount of unmet need, even among children with both a psychiatric diagnosis and functional impairment. The findings point to a significant role for the education sector, suggesting that schools may function as the de facto mental health system for children and adolescents. PMID- 7498889 TI - Utah's Prepaid Mental Health Plan: the first year. AB - This DataWatch analyzes the effect of the Utah Prepaid Mental Health Plan (UPMHP) on use of mental health services by and mental health treatment expenditures for Medicaid beneficiaries from July 1991 to June 1992. During this period three community mental health centers (CMHCs) provided mental health services to Medicaid beneficiaries in their catchment areas in return for capitated payments. Utilization and expenditure rates per beneficiary per month were analyzed using a "fixed-effects" statistical modeling approach, controlling for categories of beneficiary, time trends, seasonal effects, and CMHC grouping (capitated urban, capitated rural, noncapitated urban, and noncapitated rural). The results of the analysis suggest that the UPMHP reduced admissions for inpatient mental health treatment, inpatient mental health expenditures, and total mental health expenditures for Medicaid beneficiaries. These findings must be regarded as preliminary because of the relatively short time period covered by the data. PMID- 7498890 TI - Mental health/substance abuse treatment in managed care: the Massachusetts Medicaid experience. AB - Massachusetts was the first state to introduce a statewide specialty mental health managed care plan for its Medicaid program. This study assesses the impact of this program on expenditures, access, and relative quality. Over a one-year period, expenditures were reduced by 22 percent below predicted levels without managed care, without any overall reduction in access or relative quality. Reduced lengths-of-stay, lower prices, and fewer inpatient admissions were the major factors. However, for one population segment--children and adolescents- readmission rates increased slightly, and providers for this group were less satisfied than they were before managed care was adopted. Less costly types of twenty-four-hour care were substituted for inpatient hospital care. This experience supports the usefulness of a managed care program for mental health and substance abuse services, and the applicability of such a program to high risk populations. PMID- 7498891 TI - State risk pools and mental health care use. AB - State risk pools provide an opportunity for persons with mental health and substance abuse (MH/SA) problems to purchase health insurance. This study uses data from eight risk pools during the period 1988-1991 to analyze the utilization and enrollment experience for persons who submit claims for MH/SA treatment. Special consideration is given to the effect of variation in inpatient benefits across risk pools. The experience of Connecticut's risk pool differs markedly from that of the other risk pools. Given that two states (Connecticut and Florida) have restricted MH/SA benefits over time, we discuss the ability of risk pools to maintain comprehensive MH/SA benefits. PMID- 7498892 TI - Toward managed care for persons with severe mental illness: implications from a cost-effectiveness study. AB - Over the past two decades various models of community-based care for persons with severe mental illness have been developed. This study, which represents the first comparison of the most successful care approach (Program of Assertive Community Treatment or PACT adaptation model) with other, less intensive approaches (clinical team and intensive broker models) in a community service system, indicates that client outcomes were more positive in the PACT adaptation model in terms of enhanced psychosocial functioning and reduced acute and subacute care costs. The PACT model was not significantly more expensive in terms of the costs of providing supportive services compared with the clinical team approach and the intensive broker model of care. PMID- 7498893 TI - Partial capitation versus fee-for-service in mental health care. AB - Reimbursement strategies represent a key tool in implementing public policy. Often the relationship between changes in reimbursement policy and changes in provider behavior is difficult to predict, particularly in complex service systems. Here we present findings from a multiyear study involving the comparison of alternative Medicaid reimbursement systems for intensive case management services: a fee-for-service arrangement developed specifically for this study, and the partial capitation system in use in New York State. We focus on the impact of these two reimbursement schemes on the number and types of services provided by case managers and on client outcomes. PMID- 7498894 TI - A capitated payment system for involuntary mental health clients. AB - This DataWatch examines an outpatient capitated payment system, in the state of Oregon, designed to enhance community services for persons with chronic mental illness who had repeatedly been hospitalized involuntarily. Special state funds and Medicaid dollars were used to pay providers prospectively on a risk-adjusted basis for the delivery of outpatient mental health services. During the three year study period clients were able to be discharged from the state hospital. Although the data are not straightforward, capitated clients' use of the state mental hospital seems to have declined somewhat more than that of comparison subjects. Outpatient service use was modest and appeared to have little relationship to a client's level of illness severity. Indeed, it was not possible to predict prospectively these clients' outpatient mental health services expenditures. PMID- 7498896 TI - The physician workforce delusion. PMID- 7498895 TI - Decategorizing health services: interim findings from the Robert Wood Johnson Foundation's Child Health Initiative. AB - Although results from the evaluation are preliminary thus far, certain tentative conclusions can be reached. First, both care coordination on a small scale and the production of community health report cards are achievable within the relatively short life of a foundation grant. Moreover, both efforts can result in tangible improvements for children and their families. Report cards associated with the initiative have made children's issues more prominent and appear to have led many community leaders to focus greater attention on children's needs. Likewise, many of the care coordination systems developed under the initiative have produced real change for children and their families by guiding them to needed health care and other services. It is important, however, to keep these accomplishments in perspective. While of significant benefit to demonstration communities, the monitoring and care coordination components of this initiative are not unique. A large number of communities have adopted monitoring and reporting programs in recent years. Similarly, care coordination efforts are well established in many communities. What is unique about the RWJF initiative is its attempt at decategorization, and much less progress has been demonstrated for this component. The less-than-hoped-for progress in implementing decategorization at the original sites appears to be the product of a number of interrelated factors. These include an absence of existing models and appropriate technical assistance; political difficulties in gaining cooperation from multiple local agencies involved in service provision; limited progress in establishing needed connections with the state and federal agencies that have authority over categorical programs; and difficulties in implementing major programatic changes when the health care system itself is undergoing rapid change. In combination, these barriers have proven to be largely insurmountable for the originally funded sites, although it is too early in the project to determine which of these factors is predominantly responsible for the lack of success. Whether the newer sites can learn from the experience of the first group and adapt strategies to overcome the multiple hurdles involved remains to be seen. Decategorization is a tool that has the potential to rationalize a fragmented service system by facilitating the coordination of services, especially for children and families with multiple needs. The need for decategorization of funds will not disappear, even if the federal government chooses to combine more of its grant programs to the states into block grants.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 7498897 TI - Physician supply for the next generation: deluge or deprivation? PMID- 7498898 TI - The role of nurses in the new health care marketplace. PMID- 7498899 TI - Balancing the health care workforce. PMID- 7498900 TI - The vagaries of making distributional assumptions. PMID- 7498901 TI - States and technology assessment. PMID- 7498902 TI - States' embrace of managed mental health care. AB - Historically, state mental health authorities have dominated public mental health services, operating with fixed resources and responsible for a large population. A good public mental health system has many of the attributes of a well-managed private mental health system. Unfortunately, public systems are not flexible enough to contract creatively with multiple providers; they lack many of the tools of modern managed care. As a consequence, state mental health authorities have begun to contract with private managed care firms to assist them in managing their health care reform efforts, particularly reform of Medicaid. This paper examines the forces shaping managed behavioral health care in the public sector and describes strategies for managing care, such as contracting, utilization review, and monitoring. PMID- 7498903 TI - Risk contracts in managed mental health care. AB - Private employers and state Medicaid programs are increasingly writing risk contracts with managed behavioral health care companies to manage mental health and substance abuse benefits. This paper analyzes the case for a carve-out program and makes recommendations about the form of the payer-managed behavioral health care contract. Payers should consider using a "soft" capitation contract in which only some of the claims' risk is transferred to the managed behavioral health care company. To avoid incentives to underserve seriously ill persons, we recommend that payers not allow choice by enrollees among risk contractors. PMID- 7498904 TI - Managed care and the role and training of psychiatrists. AB - While each sector of medical practice and academic medicine confronts the reality of a changing economic environment driven by managed care, psychiatry faces some of the most difficult challenges in defining the future roles and training of psychiatrists. In this paper we describe the challenges and opportunities for psychiatry in this new era, as well as some of the unique problems facing academic psychiatry departments as they seek to fund their academic mission. PMID- 7498905 TI - Managed care in mental health: the ethical issues. AB - Praise and blame of managed mental health care are on the rise on many fronts, including allegations that it could adversely affect quality of care, access to care, the physician/patient relationship, and informed patient choice. Given the heterogeneity among managed mental health care organizations--each with differing practices--it is difficult to sift the ethically defensible concerns from the indefensible ones. In this paper we identify and examine the different moral concerns about managed mental health care and mark which problems have been addressed or are in need of resolution. We also identify which problems are unique to managed mental health care. PMID- 7498906 TI - Care for depression in a changing environment. AB - The U.S. health care system is quickly changing, but is it moving in the right direction? Focusing on care for clinical depression as a test case, this paper summarizes our previously published findings on the effects of various payment strategies, managed care, and primary care gatekeepers on the outcomes and costs for the treatment of mental health conditions. We then synthesize the policy implications of these findings for achieving value of care, lower costs, and good health outcomes. PMID- 7498907 TI - Measuring quality of life in a reformed health system. AB - "Quality of life" encompasses functional status, access to resources and opportunities, and sense of well-being. It offers a useful perspective on the value of health care, especially for chronically disabling conditions, including chronic mental illness. At least three major quality-of-life outcome assessment frameworks are available: general quality of life, health-related quality of life, and disease-specific quality of life. Choice of a framework must be driven by the intent of the services. For persons with chronic, disabling conditions, a general quality-of-life perspective is most appropriate, one that accounts not only for direct health outcomes but also for the potential social and economic effects of medical disability. PMID- 7498908 TI - A theoretical analysis of three-dimensional eye position measurement using polar cross-correlation. AB - Polar cross-correlation is a commonly used technique for determination of torsional eye position from video images. At eccentric eye positions, the projection of the sampling window onto the image plane of the camera is translated and deformed due to the spherical shape of the eyeball. In this paper, we extend the polar cross-correlation technique by developing the formulas required to determine the correct location and shape of the sampling window at all eye positions. These formulas also allow the representation of three dimensional eye position in Fick-angles, which are commonly used in oculomotor research. A numerical simulation shows the size of the errors in ocular torsion if the spherical geometry of the eye is not considered. Other effects which can affect the accuracy of video-based eye position measurements are also discussed. PMID- 7498909 TI - Self-organizing map in recognition of topographic patterns of EEG spectra. AB - The self-organizing map, a neural network algorithm, was applied to the recognition of topographic patterns in clinical 22-channel EEG. Inputs to the map were extracted from short-time power spectra of all channels. Each location on a self-organized map entails a model for a cluster of similar input patterns; the best-matching model determines the location of a sample on the map. Thus, an instantaneous topographic EEG pattern corresponds to the location of the sample, and changes with time correspond to the trajectories of consecutive samples. EEG segments of "alpha," "alpha attenuation," "theta of drowsiness," "eye movements," "EMG artifact," and "electrode artifacts" were selected and labeled by visual inspection of the original records. The map locations of the labeled segments were different; the map thus distinguished between them. The locations of individual EEG's on the "alpha-area" of the map were also different. The clustering and easily understandable visualization of topographic EEG patterns are obtainable on a self-organized map in real time. PMID- 7498911 TI - Iterative Principal Axes Registration method for analysis of MR-PET brain images. AB - Computerized automatic registration of MR-PET images of the brain is of significant interest for multimodality brain image analysis. In this paper, we discuss the Principal Axes Transformation for registration of three-dimensional MR and PET images. A new brain phantom designed to test MR-PET registration accuracy determines that the Principal Axes Registration method is accurate to within an average of 1.37 mm with a standard deviation of 0.78 mm. Often the PET scans are not complete in the sense that the PET volume does not match the respective MR volume. We have developed an Iterative Principal Axes Registration (IPAR) algorithm for such cases. Partial volumes of PET can be accurately registered to the complete MR volume using the new iterative algorithm. The quantitative and qualitative analyses of MR-PET image registration are presented and discussed. Results show that the new Principal Axes Registration algorithm is accurate and practical in MR-PET correlation studies. PMID- 7498910 TI - Three-dimensional anatomical model-based segmentation of MR brain images through Principal Axes Registration. AB - Model-based segmentation and analysis of brain images depends on anatomical knowledge which may be derived from conventional atlases. Classical anatomical atlases are based on the rigid spatial distribution provided by a single cadaver. Their use to segment internal anatomical brain structures in a high-resolution MR brain image does not provide any knowledge about the subject variability, and therefore they are not very efficient in analysis. We present a method to develop three-dimensional computerized composite models of brain structures to build a computerized anatomical atlas. The composite models are developed using the real MR brain images of human subjects which are registered through the Principal Axes Transformation. The composite models provide probabilistic spatial distributions, which represent the variability of brain structures and can be easily updated for additional subjects. We demonstrate the use of such a composite model of ventricular structure to help segmentation of the ventricles and Cerebrospinal Fluid (CSF) of MR brain images. In this paper, a composite model of ventricles using a set of 22 human subjects is developed and used in a model-based segmentation of ventricles, sulci, and white matter lesions. To illustrate the clinical usefulness, automatic volumetric measurements on ventricular size and cortical atrophy for an additional eight alcoholics and 10 normal subjects were made. The volumetric quantitative results indicated regional brain atrophy in chronic alcoholics. PMID- 7498912 TI - The use of arch index to characterize arch height: a digital image processing approach. AB - Attempts to evaluate foot arch types from footprint parameters have yielded conflicting results in the past. This could be caused by the uncertainty inherent in the definition of some footprint parameters and the inaccuracy during the footprint acquisition and the parameter calculation phases of the traditional methods. In order to avoid these problems, digital image processing methods were used to acquire and to calculate the Arch Index (AI), a parameter which is robust in its definition. A significant correlation (r = -0.70, p < 0.0001) was found between AI and arch height. Therefore this study confirms that foot arch type does correlate with the footprint parameter, AI. This was further revealed by a new parameter, the Modified Arch Index (MAI), which incorporates foot pressure information in the evaluation. MAI not only correlated well with arch height (r = -0.71, p < 0.0001) but appeared to characterize abnormal foot types better than AI. PMID- 7498913 TI - Improved method for computation of potentials in a realistic head shape model. AB - The Lead Field Analysis (LFA) algorithm, a new computational technique for the calculation of potentials on the surface of a realistic head shaped volume conductor model based on the Boundary Element Method and the Reciprocity Theorem, is presented. The new algorithm, in comparison to the Standard Boundary Element Method, offers improved computational efficiency and lower storage requirements. It also yields more accurate surface potential results in the face of varying dipole source locations for a head shape Boundary Element model with a given number of nodes. Additionally, the algorithm results in quasi-analytic expressions of the derivatives of the surface potential with respect to the location of the sources, allowing the use of optimization techniques with better convergence properties. A set of simulations demonstrating the increased robustness of the LFA Algorithm in the face of varying dipole source parameters is also described. PMID- 7498914 TI - Transmission-line electric field induction in humans using charge simulation method. AB - This paper is aimed at determining the distribution of the fields, induced charges, and currents on a human body standing in the high electric fields produced by high voltage overhead transmission lines. This method of analysis is based on the charge simulation technique. This will serve to explain the biological studies of possible long-term exposure effects to electric fields. PMID- 7498916 TI - Neural network control of functional neuromuscular stimulation systems: computer simulation studies. AB - A neural network control system has been designed for the control of cyclic movements in Functional Neuromuscular Stimulation (FNS) systems. The design directly addresses three major problems in FNS control systems: customization of control system parameters for a particular individual, adaptation during operation to account for changes in the musculoskeletal system, and attaining resistance to mechanical disturbances. The control system was implemented by a two-stage neural network that utilizes a combination of adaptive feedforward and feedback control techniques. A new learning algorithm was developed to provide rapid customization and adaptation. The control system was evaluated in a series of studies on a computer simulated musculoskeletal model. The model of electrically stimulated muscle used in the study included nonlinear recruitment, linear dynamics, and multiplicative nonlinear torque-angle and torque-velocity scaling factors. The skeletal model consisted of a one-segment planar system with passive constraints on joint movement. Results of the evaluation have demonstrated that the control system can provide automated customization of the feedforward controller parameters for a given musculoskeletal system. It can account for changes in the musculoskeletal system by adapting the feedforward controller parameters on-line and it can resist the effects of mechanical disturbances. These results suggest that this design may be suitable for the control of FNS systems and other dynamic systems. PMID- 7498915 TI - Magnetically induced currents in the canine heart: a finite element study. AB - A moderately detailed three-dimensional (3-D) finite element model of the conductive anatomy of a canine thorax was used to determine the fields and currents induced by a time-varying magnetic field that has been shown to cause irregular heart beats in canines. The 3-D finite element model of the canine thorax was constructed from CT scans and includes seven isotropic tissue conductivities and the anisotropic conductivity of skeletal muscle. We use this model to estimate the stimulation threshold associated with stimulation of the heart by the time-varying magnetic field of a figure-eight coil. Variants of the thoracic model were also constructed to examine the sensitivity of model results to variations in model size, shape, and conductive inhomogeneity and anisotrophy. Our results show that myocardial fields were only midly sensitive to thoracic size. However, model shape and conductive inhomogeneity and anisotrophy substantially influenced the magnitude and distribution of myocardial fields and currents. Our results suggest that an induced peak field magnitude of approximately 1 V/cm is required to stimulate the heart with the magnetic excitation simulated in this study. PMID- 7498917 TI - Elimination of AC interference in electrocardiogram using IIR notch filter with transient suppression. AB - In this paper, a technique for suppressing the transient states of IIR notch filter is investigated. This technique uses the vector projection to find better initial values for notch filters. When a notch or comb filter is used to eliminate power line (AC) interference in the recording of electrocardiograms (ECG), the performance of the notch filter with transient suppression is better than that of the conventional notch filter with arbitrary initial condition. The improvements with this technique are at the cost of additional computation load at the beginning of filtering. PMID- 7498918 TI - Signal-to-noise ratio enhancement of cardiac late potentials using ensemble correlation. AB - An optimal one-weight "filter" is presented for the purpose of improving the signal-to-noise ratio of averaged ECG recordings in the analysis of late potentials. Based on a simple statistical model, the filter is estimated from the ensemble correlation of available beats. The correlation estimator is found by a maximum likelihood procedure in which the observed signal is assumed to have a Gaussian distribution. The performance of the optimal filter is studied in relation to an ensemble with individual or subaveraged beats. PMID- 7498920 TI - A new method for approximation of physiological signals. AB - A new robust method for the approximation of physiological signals is proposed. The noise must satisfy only one condition: that the probabilities for positive and negative values are equal. Our results illustrate that the proposed method provides a more accurate approximation of signals than either the least square error or the least absolute deviation method in cases that include unknown noise density. PMID- 7498919 TI - Genetic design of optimum linear and nonlinear QRS detectors. AB - This paper describes an approach to the design of optimum QRS detectors. We report on detectors including a linear or nonlinear polynomial filter, which enhances and rectifies the QRS complex, and a simple, adaptive maxima detector. The parameters of the filter and the detector, and the samples to be processed are selected by a genetic algorithm which minimizes the detection errors made on a set of reference ECG signals. Three different architectures and the experimental results achieved on the MIT-BIH Arrhythmia Database are described. PMID- 7498922 TI - Stuart Downs. Interview by Carla McGlone. PMID- 7498921 TI - Amber Rea. Interview by Carla McGlone. PMID- 7498923 TI - The Immigration Nursing Relief Act of 1989. PMID- 7498924 TI - ANA launches "every patient deserves a nurse" campaign. PMID- 7498925 TI - Flight nursing: a view from above. PMID- 7498927 TI - The challenge of emergency nursing. PMID- 7498926 TI - Perioperative nursing. PMID- 7498928 TI - Home health nursing. PMID- 7498929 TI - Rehabilitation nursing. PMID- 7498930 TI - "University hospital" nothing like real life. PMID- 7498932 TI - Let the caring show. PMID- 7498931 TI - Sharing: my world and theirs. PMID- 7498933 TI - Running toward the pain. PMID- 7498934 TI - The Immigration Nursing Relief Act of 1989: Part II. PMID- 7498935 TI - Staci Deck. Interview by Carla McGlone. PMID- 7498936 TI - Iris Antcliff. Interview by Carla McGlone. PMID- 7498938 TI - Enriching the HIV/AIDS education of baccalaureate nursing students through independent study. PMID- 7498937 TI - Meeting the challenge of HIV/AIDS. PMID- 7498939 TI - Women and HIV/AIDS. PMID- 7498940 TI - Profile: the International Center for HIV/AIDS Research and Clinical Training in Nursing. PMID- 7498941 TI - "Why I chose nursing" a letter to mom. PMID- 7498942 TI - Disaster relief: perspectives from nursing students in Oklahoma City. PMID- 7498943 TI - Tragedy in the heartland area nursing students and faculty unite in Oklahoma. PMID- 7498944 TI - Study tips from and for nursing students. PMID- 7498945 TI - Tips for stress management. PMID- 7498946 TI - Student to student. PMID- 7498948 TI - 1995 publishers directory. Resource books for nursing students. PMID- 7498947 TI - How to survive transfer trauma. PMID- 7498949 TI - Should more men be recruited into nursing? PMID- 7498950 TI - Gene Cagle. PMID- 7498951 TI - Cultural considerations: Asian-Americans and Pacific Islanders. PMID- 7498952 TI - Students experience the Pine Ridge South Dakota Reservation. PMID- 7498953 TI - Cultural considerations: providing quality nursing care to Hispanics. PMID- 7498954 TI - Cultural considerations for the African-American population. PMID- 7498956 TI - "Why I chose nursing as a career". PMID- 7498955 TI - ANA and NSNA join forces against breast cancer. PMID- 7498957 TI - A nursing student experiences "hands-on" community health nursing. PMID- 7498958 TI - Teaching a client with low literacy skills. PMID- 7498959 TI - Saved by her hair. PMID- 7498960 TI - Effects of long-term nitric oxide synthesis inhibition on plasma volume expansion and fetal growth in the pregnant rat. AB - We conducted the present study to investigate whether the vasodilator nitric oxide plays a role in plasma volume homeostasis during pregnancy. Pregnant Sprague-Dawley rats were randomly assigned to a control group (n = 18) or to groups receiving 0.69 mmol/L (n = 11) or 1.7 mmol/L (n = 14) N omega-nitro-L arginine, a competitive inhibitor of nitric oxide synthetase, from gestational days 7 through 21. On day 20 systolic pressure was measured. On day 21 blood samples were taken for plasma volume, hematocrit, and hormonal measurements. Fetal and placental weights also were determined. Systolic pressure was significantly higher in experimental rats (101 +/- 6 and 115 +/- 6 mm Hg in the 0.69 and 1.7 mmol/L groups, respectively) than in controls (79.7 +/- 7.5 mm Hg), and plasma volume was lower (18.4 +/- 1.1 and 17.1 +/- 0.5 mL) than in controls (21.5 +/- 0.8 mL). Both experimental groups had increased hematocrit levels. Plasma renin activity was significantly lower in the experimental groups (11.5 +/ 3 and 7.2 +/- 1.5 ng angiotensin I/mL per hour) than in controls (21.9 +/- 2.7 ng angiotensin I/mL per hour); however, no changes were observed in aldosterone levels. Experimental groups had lower fetal weight (4.6 +/- 0.1 and 5.1 +/- 0.1 g) than controls (5.5 +/- 0.1 g). In addition, fetal hindlimb hypoplasia was observed in the experimental groups. In conclusion, the present data indicate that long-term N omega-nitro-L-arginine administration to pregnant rats leads to increased blood pressure, reduced plasma volume expansion, lower plasma renin activity, and fetal growth retardation.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7498961 TI - Role of nitric oxide in modulating the vasoconstrictor actions of angiotensin II in preglomerular and postglomerular vessels in dogs. AB - The purpose of this study was to determine the role of nitric oxide in modulating the vasoconstrictor effect of angiotensin II (Ang II) on renal segmental resistances in the dog. To achieve this objective we examined the effect of intrarenal infusions of Ang II on preglomerular and postglomerular resistances in the presence and absence of intrarenal nitric oxide synthesis inhibition established by an intrarenal infusion of NG-nitro-L-arginine-methyl ester at 5 micrograms/kg per minute in dogs. The whole-kidney stop-flow technique was used. Renal artery pressure was servo-controlled at 78 +/- 2 mm Hg throughout the study. Intrarenal infusion of Ang II alone at 0.5 and 2.0 ng/kg per minute increased renal vascular resistance (delta 0.064 +/- 0.011 and delta 0.171 +/- 0.030 mm Hg/mL per minute, respectively) and decreased renal blood flow (delta 21 +/- 4 and delta 45 +/- 9 mL/min). Associated with these changes, glomerular hydrostatic pressure and preglomerular resistance increased slightly (delta 1.1 +/- 0.9 and delta 1.6 +/- 1.8 mm Hg; delta 0.008 +/- 0.005 and delta 0.030 +/- 0.010 mm Hg/mL per minute, respectively), and postglomerular resistance increased markedly (delta 0.046 +/- 0.011 and delta 0.116 +/- 0.026 mm Hg/mL per minute). When dogs were pretreated with an intrarenal infusion of the nitric oxide synthesis blocker, Ang II at 0.5 and 2.0 ng/kg per minute increased renal vascular resistance (delta 0.271 +/- 0.058 and delta 1.088 +/- 0.242 mm Hg/mL per minute) and decreased renal blood flow (delta 28 +/- 5 and delta 62 +/- 9 mL/min).(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7498962 TI - Nitric oxide synthase isoform activities in kidney of Dahl salt-sensitive rats. AB - An abnormal L-arginine-nitric oxide axis has been suggested to be relevant to the genesis of salt-sensitive hypertension. In the present study we investigated the activities of three isoforms of nitric oxide synthase (NOS) in the kidney of Dahl salt-sensitive and salt-resistant rats. Five-week-old Dahl Iwai salt-sensitive (n = 9) and salt-resistant (n = 10) rats were maintained on a high salt diet (4% sodium chloride) for 4 weeks. We measured calcium-dependent and calcium independent NOS activities in each particulate and soluble fraction of kidney by conversion of L-[3H]arginine to L-[3H]citrulline. Systolic blood pressure was elevated significantly (P < .001) in salt-sensitive but not salt-resistant rats. Calcium-dependent NOS activity in the soluble fraction was significantly lower in salt-sensitive rats than in salt-resistant rats (25.8 +/- 9.0 versus 48.2 +/- 19.2 disintegrations per microgram protein, respectively; P < .01). There were no differences in calcium-dependent NOS activity in the particulate fraction and calcium-independent NOS activity in the soluble fraction between groups. Renal norepinephrine content was lower in salt-sensitive rats than in salt-resistant rats (P < .05) and was positively correlated with calcium-dependent NOS activity in the soluble fraction (P < .01). Although no differences in endothelial and inducible-type NOS activity were observed a significant reduction in calcium dependent NOS activity in the soluble fraction of the kidney of salt-sensitive rats suggests that the decreased neural-type NOS activity may in part be involved in the mechanism of salt-sensitive hypertension, possibly through alterations in renal sympathetic nervous activity and sodium handling. PMID- 7498963 TI - Prostacyclin synthesis elicited by endothelin-1 in rat aorta is mediated by an ETA receptor via influx of calcium and is independent of protein kinase C. AB - The purpose of this study was to characterize the receptor(s) and second messenger systems involved in prostacyclin (prostaglandin [PG] I2) synthesis elicited by endothelin (ET)-1 in the rat aorta. PGI2 synthesis, measured as immunoreactive 6-keto-PGF1 alpha, was assessed in aortic rings exposed to endothelin receptor agonists in the presence and absence of selective ETA and ETB receptor antagonists. ET-1, which has equal affinity for both endothelin receptor subtypes, and ET-3, a preferential ETB receptor agonist, enhanced 6-keto-PGF1 alpha synthesis in a time- and concentration-dependent manner. ET-1 was more potent than ET-3 in increasing 6-keto-PGF1 alpha synthesis. Moreover, the selective ETB receptor agonists IRL-1620 and sarafotoxin S6c did not significantly increase 6-keto-PGF1 alpha synthesis. Furthermore, ET-1-induced 6 keto-PGF1 alpha synthesis was attenuated by an ETA receptor antagonist, BQ-123, in a dose-dependent manner but not by an ETB receptor antagonist, BQ-788. Depletion of extracellular Ca2+ or addition of Ca2+ channel blockers (nifedipine, verapamil, SK&F 96365) attenuated ET-1-mediated 6-keto-PGF1 alpha synthesis, while a Ca2+ channel agonist, S(-)-Bay K 8644, potentiated this effect of ET-1. Selective protein kinase C inhibitors (bisindolylmaleimide I, calphostin C) did not alter ET-1-induced 6-keto-PGF1 alpha synthesis. These data suggest that PGI2 synthesis elicited by ET-1 in the rat aorta is mediated primarily through influx of extracellular Ca2+ via activation of an ETA receptor and is independent of protein kinase C. PMID- 7498964 TI - Endothelin subtype B receptor-mediated calcium and contractile responses in small arteries of hypertensive rats. AB - Endothelin-1 elicits vasoconstrictor responses through endothelin subtype A receptors, which are located on vascular smooth muscle cells, and vasodilator responses through endothelin subtype B receptors, which occur predominantly on endothelial cells. Endothelin subtype B receptors also may be present on vascular smooth muscle cells, in which they may mediate vasoconstriction. The aims of this study were to determine the presence of vascular smooth muscle vasoconstrictor endothelin subtype B receptors in mesenteric resistance arteries and to assess whether endothelin subtype B receptor-mediated responses differ between spontaneously hypertensive rats and Wistar-Kyoto rats. Contractile responses to the endothelin subtype B receptor agonist sarafotoxin S6c and endothelin-1 were measured simultaneously with [Ca2+]i in endothelium-denuded mesenteric resistance arteries from adult spontaneously hypertensive rats and Wistar-Kyoto rats. To simulate in vivo conditions matched as closely as possible to in vitro conditions, vessels were mounted in a vessel flow chamber in which intraluminal pressure was maintained at 60 mm Hg. Contraction was determined by video imaging to record lumen diameter, and [Ca2+]i was measured by the fura 2 method. Basal [Ca2+]i was significantly higher (P < .01) in hypertensive (170 +/- 4 nmol/L) compared with normotensive rats (134 +/- nmol/L). The endothelin subtype B receptor agonist sarafotoxin S6c increased [Ca2+]i in a concentration-dependent manner. Sarafotoxin S6c-induced [Ca2+]i and contractile responses were significantly lower in hypertensive compared with normotensive rats. These data demonstrate that endothelin subtype B receptors are present in vascular smooth muscle of small arteries and that endothelin subtype B receptor-mediated vasoconstriction occurs through intracellular calcium signaling pathways.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7498965 TI - Heterogeneity of adenovirus-mediated gene transfer in cultured thoracic aorta and renal artery of rats. AB - Replication-deficient recombinant adenovirus vectors have been used to transfer foreign genes effectively to a wide variety of cell types in vivo and in vitro. We have now used adenovirus containing either the Escherichia coli beta galactosidase (beta-gal) gene (AdHCMVsp1LacZ) or the firefly luciferase gene (Ad5 luc3) to test the hypothesis that efficiencies of adenovirus-mediated gene delivery into organ cultures of smooth muscle differ according to the anatomic origin of the muscle. Thoracic aorta and renal artery were isolated from 9-week old male Sprague-Dawley rats and exposed to adenovirus after 16 hours of incubation with serum-free medium (Dulbecco's modified Eagle's medium). With the use of histochemical methods, beta-gal staining was noted in both endothelial and adventitial cells but not in the muscular media of thoracic aorta and renal artery exposed to AdHCMVsp1LacZ. The efficiency of the transfection, assessed either by counting of beta-gal-stained cells in intact vessels or by measurement of beta-gal activity in tissue extracts, was higher in renal artery than thoracic aorta (P < .05). Consistent with this result, luciferase activity in renal artery exposed to Ad5-luc3 (15.9 +/- 2.1 x 10(6) relative light units per milligram protein) was higher than that in thoracic aorta (8.3 +/- 2.0 x 10(6), P < .05). To determine whether increased efficiency of adenovirus-mediated gene transfer into renal artery is a function of the replication status of vessels, we assessed [3H]thymidine incorporation. [3H]Thymidine uptake by thoracic aorta was only 63% of that in renal artery (P < .05), indicating that more proliferating cells are present in renal artery. We conclude that the efficiency of adenovirus-mediated gene transfer into cultured renal artery is enhanced compared with that into thoracic aorta and propose that the increase in efficiency is related to the higher proliferative activity of renal artery. PMID- 7498966 TI - Intracellular Ca2+ release in flow-induced contraction of venous smooth muscle. AB - We designed the present study to determine whether Ca2+ release from intracellular stores contributes to flow-induced contraction. We carried out experiments on segments of rabbit facial vein under isometric conditions. Intraluminal flow by perfusion of physiological salt solution (10 to 80 microL/min) caused contraction in this vessel, which was significantly inhibited by (1) 30-minute pretreatment with 10 mumol/L ryanodine, the sarcoplasmic reticulum Ca2+ channel opener, and (2) 30-minute pretreatment with concomitant application of 20 mmol/L caffeine and 1 mumol/L cyclopiazonic acid in Ca(2+)-free medium to deplete the sarcoplasmic reticulum. In comparison, contraction initiated by 300 nmol/L histamine was significantly attenuated by the same interventions. K+ (25 mmol/L)-induced contraction was unaffected by ryanodine but was reduced after depletion of the sarcoplasmic reticulum. The phospholipase C inhibitor 2-nitro-4-carboxyphenyl-N,N-diphenylcarbamate (10 mumol/L) inhibited contractions induced by flow and histamine but not by K+. These findings indicate that Ca2+ release from intracellular stores, presumably via the phosphatidylinositol pathway, contributes to flow- and histamine- but not raised K(+)-induced contractions in this vessel. PMID- 7498968 TI - Augmented calcium currents in mesenteric artery branches of the spontaneously hypertensive rat. AB - The greater efficacy of organic channel blockers in lowering peripheral resistance and blood pressure in hypertensive subjects has been suggested to be the result of augmented calcium influx through L-type calcium channels in arterial smooth muscle. These studies were performed to determine whether differences exist in voltage-gated calcium channels of mesenteric artery branches from 20-week-old spontaneously hypertensive rats (SHR) compared with Wistar-Kyoto rats (WKY). Single myocytes were acutely isolated by collagenase and elastase treatment and studied at room temperature (approximately 20 degrees C) with the use of whole-cell, patch-clamp methods. Maximum values of calcium current measured at 0 mV from a holding potential of -90 mV were larger in SHR myocytes (105 +/- 11 versus 149 +/- 15 pA). Values of cell capacitance were smaller in SHR (29.5 +/- 1.3 pF) compared with WKY (35.0 +/- 1.5 pF) myocytes. Cell capacitance measures surface membrane area and, when used to normalize calcium currents, magnified the difference between WKY and SHR to approximately 47%. There was a larger percent reduction of maximum calcium current at holding potentials of -60 and -40 mV in SHR compared with WKY myocytes: for example, at -40 mV calcium current was reduced from values at -90 mV by -73 +/- 2% in SHR compared with -58 +/- 1% in WKY. When divided by the maximum current for each holding potential, the voltage dependence of normalized calcium currents for the two groups was completely superimposed. Difference currents were calculated by subtracting currents measured from holding potentials of -90 and -40 mV. The voltage dependence of difference currents was identical to that of the calcium currents measured from the two values of holding potential.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7498967 TI - 5-Hydroxytryptamine2B receptor mediates contraction in the mesenteric artery of mineralocorticoid hypertensive rats. AB - Vascular responsiveness to 5-hydroxytryptamine (5-HT) is dramatically increased in hypertension. The hypothesis that augmented vasoconstriction to 5-HT in hypertension is due to a change in receptor subtype on vascular myocytes was tested. Mesenteric arteries from deoxycorticosterone acetate (DOCA)-salt hypertensive (systolic blood pressure > 180 mm Hg) and sham normotensive (systolic blood pressure < 130 mm Hg) rats were mounted in isolated tissue baths for measurement of isometric contractile force. The receptor mediating contraction in isolated mesenteric arteries from sham and DOCA-salt hypertensive rats is a member of the 5-HT2 family based on rank order of agonist potency (5-HT = alpha-methyl-5-HT [5-HT2 receptor agonist]>tryptamine>5-hydroxykynuramine). 5 HT was approximately 10-fold more potent in contracting mesenteric arteries from DOCA-salt hypertensive rats compared with arteries from sham normotensive rats. The tryptophan metabolite kynuramine, which possesses significant contractile activity at the 5-HT2B receptor, contracted hypertensive arteries significantly (50% of 5-HT maximum) but not sham arteries. Ketanserin (5-HT2A antagonist) competitively inhibited contraction to 5-HT in arteries from normotensive rats ( log dissociation constant [mol/L]; pKB = 8.54) but not from hypertensive rats (pKB > 6.5). Moreover, contraction to kynuramine was not blocked by ketanserin. Thus, under normal conditions, 5-HT2A receptors mediate contraction to 5-HT. However, in DOCA-salt hypertension, ketanserin-insensitive 5-HT2 receptors, possibly 5-HT2B receptors, mediate mesenteric arterial contraction to 5-HT. PMID- 7498969 TI - Effects of dehydroepiandrosterone sulfate on cellular calcium responsiveness and vascular contractility. AB - Dehydroepiandrosterone sulfate (DHEAS) is an endogenous steroid having a wide variety of biological effects, but its physiological role remains undefined. Since an age-related decline of DHEAS corresponds to the progressive onset of atherosclerosis, cardiovascular diseases, and overall mortality, we investigated a possible protective role of DHEAS in vascular disease by studying the effects of this hormone (10(-7) to 10(-5) mol/L) on cytosolic free calcium and contractility in different in vitro vascular tissue preparations. DHEAS produced a significant, dose-dependent relaxation of isolated helical strips of rat tail artery precontracted with KCl (60 mmol/L) (89.7 +/- 18.7%, P < .01), arginine vasopressin (3 nmol/L) (27.3 +/- 7.1%, P < .01), and norepinephrine (0.1 mumol/L) (49.2 +/- 18.2%, P < .01). In isolated vascular smooth muscle cells DHEAS reversibly inhibited KCl (30 mmol/L)-induced elevations of cytosolic free calcium to 69.8 +/- 8.4% and 43.8 +/- 7.4% of the control response at 5 x 10(-7) and 5 x 10(-6) mol/L, respectively (P < .05 at both doses). These results provide evidence of a direct vascular action of DHEAS, in doses reflecting circulating levels in vivo, and suggest the possibility that these effects are mediated by modulation of intracellular calcium metabolism. We hypothesize that physiologically, DHEAS may serve to buffer vascular responsiveness to a wide variety of depolarizing and constrictor hormonal stimuli. PMID- 7498970 TI - Calcium-ATPase and insulin in adolescent offspring of essential hypertensive parents. AB - A number of abnormalities in calcium homeostasis have been reported in patients with essential hypertension. IN turn, insulin has been shown to influence the activity of the Ca(2+)-ATPase. We have previously shown that normotensive offspring of essential hypertensive individuals have an exaggerated insulin response to a glucose overload. Therefore, the aim of the present study was to evaluate basal and calmodulin-activated Ca(2+)-ATPase in red blood cells and its relationship to the insulin response during an intravenous glucose tolerance test in 27 normotensive adolescents with a family history of essential hypertension (F+) (mean age, 13.9 +/- 0.5 years) and in 10 control subjects matched for age and body mass index with no family history of hypertension (F-). The results (mean +/- SD) were as follows (mumol Pi/[mg protein/h]10(-1)): basal Ca(2+) ATPase, 4.5 +/- 1.2 in F+ and 5.1 +/- 1.6 in F- (P = NS); calmodulin-activated Ca(2+)-ATPase, 13.6 +/- 3.9 in F+ and 16.2 +/- 1.7 in F- (P < .04). The insulin area under the curve after the glucose load was 3413 +/- 1674 microU/mL per hour in F+ and 2752 +/- 928 in F- (P = NS). Calmodulin-activated Ca(2+)-ATPase showed a negative correlation with the insulin area under the curve (r = -.59, P < .005) and cholesterol levels (r = -.38, P < .03). Urinary calcium excretion was 1.82 +/ 0.9 mmol/d in F+ and 2.47 +/- 0.9 mmol/d in F- (P = NS).(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7498972 TI - Suboptimal outcome of management of metabolic cardiovascular risk factors in Hispanic patients with essential hypertension. AB - We studied outcome of management of metabolic cardiovascular risk factors in 155 randomly chosen Hispanic hypertensive patients (mean age, 63 +/- 1 years; 79% female) screened for dyslipidemia. Hypertriglyceridemia (n = 12) or high risk adjusted low-density lipoprotein cholesterol (LDL-C) (n = 89) was found in 65%. Triglycerides did not change (6.16 +/- 0.58 to 7.44 +/- 2.34 mmol/L; P = NS) over 2.2 +/- 0.5 years. Only 58 patients with high LDL-C were treated, and 8 had no follow-up lipid tests. In the other 50, LDL-C decreased by 10 +/- 3% (P < .001) over 2.8 +/- 0.2 years but attained goal in only 12. In a subset of 24 patients with extended follow-up (3.8 +/- 0.2 years), there was an initial marked decline in LDL-C, followed by a rebound to baseline levels. In 29 of 54 patients with normal LDL-C, lipid testing was markedly overused compared with recommendations. Obesity (n = 94, 61%) did not improve in those with repeated data (+0.6 +/- 0.8 kg; P = NS; n = 40) over 2.7 +/- 0.3 years. Forty-four of 63 patients with type II diabetes had repeated measurement of glycosylated hemoglobin, with no change (10.5 +/- 0.5% to 11.2 +/- 0.5%; P = NS) over 2.2 +/- 0.3 years. Ten-year risk of coronary events (Framingham cohort parametric regression) calculated for 61 patients with known untreated blood pressures (169 +/- 3/98 +/- 1 mm Hg) was 21.0 +/- 1.7%, with a skewed distribution reaching high values (66%) and attributable in large part (72%) to modifiable risk factors.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7498971 TI - Effects of losartan on blood pressure, metabolic alterations, and vascular reactivity in the fructose-induced hypertensive rat. AB - Fructose feeding induces a moderate increase in blood pressure levels in normal rats that is associated with insulin resistance, hyperinsulinemia, and hypertriglyceridemia. The sympathetic nervous system seems to participate in the alterations of this model. To further explore the mechanisms underlying fructose induced hypertension, the effects of the AT1 receptor antagonist losartan on blood pressure, insulin resistance, renal function, and vascular reactivity in mesenteric vascular beds were studied. Sprague-Dawley rats were fed for 4 weeks with diets containing 60% fructose or 60% starch (control), and half of each group received losartan (1 mg/kg per day) in the drinking water. Fructose-fed rats showed higher (P < .05) blood pressure levels and plasma concentrations of triglycerides and insulin than those of controls. Losartan treatment prevented both blood pressure elevation and hyperinsulinemia in fructose-fed rats but not elevation of plasma triglycerides. Plasma glucose and insulin levels in response to an oral glucose load were higher (P < .05) in fructose-fed rats than in controls. These exaggerated responses were prevented by losartan treatment. No differences in the constrictor responses of mesenteric vascular beds to KCl (60 mumol), angiotensin II (1 nmol), phenylephrine (10(-5) mol/L), or endothelin-1 (10 pmol) were found between the two groups. Relaxing responses to acetylcholine or sodium nitroprusside in phenylephrine-precontracted mesenteric vascular beds and constrictor response to the nitric oxide synthesis inhibitor NG-nitro-L arginine methyl ester (100 nmol) were comparable in both groups. Losartan blunted angiotensin II constriction and reduced (P < .05) responses to phenylephrine in all groups.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7498973 TI - Serum insulin levels, 24-hour blood pressure profile, and left ventricular mass in nonobese hypertensive patients. AB - In essential hypertensive patients, considered to be insulin-resistant, a blunted decline in nocturnal blood pressure is associated with increased adrenergic tone and left ventricular mass. Since insulin stimulates the sympathetic system, we tested whether insulin resistance and insulinemia influence left ventricular mass and the 24-hour blood pressure profile. We studied 29 nonobese hypertensive patients with office diastolic pressure between 95 and 110 mm Hg and normal oral glucose tolerance test after a 4-month washout period. They were then assigned to M-mode echocardiographic evaluation and 24-hour ambulatory blood pressure monitoring. The glucose and insulin responses to a 75-g oral glucose load were compared with those obtained in 16 weight-matched normotensive control subjects. During the oral glucose tolerance test the hypertensive patients compared with control subjects presented higher levels of glucose at 60 minutes (138.7 +/- 30.3 versus 108.7 +/- 35.7 mg/dL; P < .05) and 90 minutes (114.0 +/- 23.8 versus 94.8 +/- 31.1 mg/dL; P < .05) and insulin at 60 minutes (287.1 +/- 259.4 versus 142.1 +/- 83.9 pmol/L; P < .05). However, peak insulin levels after glucose load did not correlate with ambulatory blood pressure values or left ventricular mass index. Left ventricular mass index showed significant correlation with mean sleeping systolic pressure (rs = 56, P < .05) and diurnal systolic pressure (rs = .37, P < .05) but not with mean diurnal or sleeping diastolic pressures. In conclusion, our results indicate that in nonobese hypertensive patients, insulin resistance does not have any influence on the 24-hour blood pressure profile or on left ventricular mass index.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7498974 TI - Insulin effect on renal sodium reabsorption in adolescent offspring of essential hypertensive parents. AB - We previously showed that children and adolescent offspring of patients with essential hypertension have an increased proximal renal sodium reabsorption as measured by lithium fractional excretion. Insulin has been shown to have antinatriuretic properties and to be increased (hyperinsulinemia) in essential hypertension. The aim of this study was to evaluate the role of insulin on the increased proximal renal sodium reabsorption previously reported. Lithium and sodium fractional excretions were measured 3 hours before and 3 hours after an intravenous glucose tolerance test in 20 normotensive adolescents with a family history of essential hypertension (F+, 14.8 +/- 0.5 years) and 10 normotensive control subjects without a family history of hypertension (F-, 15.2 +/- 0.9 years). Results are mean +/- SEM. Lithium fractional excretion before glucose loading was 16.1 +/- 1.8% in F+ versus 23.5 +/- 2.0% in F- (P < .02) and after glucose loading was 14.7 +/- 1.3% in F+ versus 20.9 +/- 1.7% in F- (P = NS). Lithium fractional excretion did not change after intravenous glucose loading in either group. The insulin area under the curve was 2815 +/- 499 in F+ versus 2290 +/- 418 microU/mL per hour in F- (P = NS). There was no correlation between lithium fractional excretion and insulin area under the curve. Fractional excretion of sodium before glucose loading was 0.99 +/- 0.1% in F+ versus 0.99 +/ 0.1% in F- (P = NS) and after glucose loading was 0.77 +/- 0.1 in F+ versus 0.85 + 0.1% in F- (P < .01 versus values before loading in both groups).(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7498975 TI - Antihypertensive treatment efficacy in type II diabetes mellitus. Dissociation between casual and 24-hour ambulatory blood pressure. Spanish Multicenter Study Group. AB - Whole-day ambulatory blood pressure monitoring is used to confirm the diagnosis of hypertension and assess the response to antihypertensive therapy. Neither of these has been applied to patients with type II diabetes mellitus, in whom it has been proposed that the desirable blood pressure should be lower than in nondiabetics. This multicenter study was designed to examine whether there are differences in the efficacy of a first-line antihypertensive drug when assessed by casual and ambulatory blood pressure determinations in patients with type II diabetes mellitus in whom 24-hour ambulatory monitoring confirms or fails to confirm the diagnosis of hypertension. Forty-three patients (mean age, 57.7 years) with stable type II diabetes mellitus and mild hypertension (casual diastolic pressure, 90 to 104 mm Hg on at least two visits) were treated with an angiotensin-converting enzyme inhibitor (benazepril, 10 to 20 mg, once a day) for 8 weeks. Antihypertensive drug efficacy was assessed by casual (trough) and 24 hour ambulatory blood pressure monitoring. Diabetic patients were classified as nonconfirmed hypertensive if the mean 24-hour ambulatory diastolic pressure was below 85 mm Hg. Antihypertensive treatment significantly decreased both systolic and diastolic pressures when determined by either casual measurement (from a mean of 162.7/98.0 to 153.9/89.2 mm Hg; P < .001) or ambulatory monitoring (from a mean of 143.1/84.4 to 137.0/81.5 mm Hg; P < .05). Twenty-one patients (49%) were classified as confirmed hypertensive and 22 as nonconfirmed hypertensive. In confirmed hypertensive patients benazepril significantly reduced systolic and diastolic pressures when assessed by either casual or 24-hour ambulatory monitoring.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7498976 TI - Autonomic dysfunction in short-term experimental diabetes. AB - Previous data showed that diabetes induced by streptozotocin for 5 days causes changes in arterial pressure control and baroreflex regulation of heart rate in male Wistar rats. The impairment of baroreflex may be related to autonomic neuropathy as described by several investigators. The aim of this study was to identify autonomic changes in short-term experimental diabetes in rats (induced for 5 days with streptozotocin 65 mg IP). Intra-arterial blood pressure signals were obtained from 6 control group and 7 diabetic group rats and processed in a data acquisition system (CODAS, 1 kHz). Both vagal and sympathetic function were assessed through intravenous injections of methylatropine and propranolol. Streptozotocin induced hyperglycemia (18.9 +/- 1.8 versus 5.8 +/- 0.2 mmol/L) and reductions in mean arterial pressure (102 +/- 2 versus 117 +/- 3 mm Hg) and resting heart rate (298 +/- 14 versus 332 +/- 2 beats per minute). Sodium and potassium levels were not different between groups. The intrinsic heart rate was reduced in the diabetic group (302 +/- 10 versus 398 +/- 6 beats per minute). This group also exhibited depressed vagal and sympathetic tone (50% and 22%, respectively), reduction of vagal effect (42%), and no change in sympathetic effect. In conclusion, early autonomic dysfunction in short-term streptozotocin induced diabetes seems to be related to changes in arterial pressure and baroreflex control. PMID- 7498977 TI - Thyrotropin-releasing hormone hyperactivity in the preoptic area of spontaneously hypertensive rats. AB - Thyrotropin-releasing hormone (TRH) plays an important role in central cardiovascular regulation through the activation of different neurotransmitter systems at distinct extrahypothalamic sites. To study possible alterations in the TRH system in the hypertensive state, we measured TRH concentration in cerebrospinal fluid and TRH content of the preoptic area in spontaneously hypertensive rats (SHR) and Wistar-Kyoto rats (WKY) by radioimmunoassay. In addition, we also measured the density of the TRH receptor in this area by a rapid filtration technique using [3H]methyl-TRH. We found a significant increase in both the TRH content (634 +/- 61 versus 350 +/- 26 pg/mg protein, SHR versus WKY; P < .01, n = 5) and density of TRH receptors without changes in affinity (Bmax, 5.0 +/- 0.1 versus 3.3 +/- 0.1 fmol/mg protein, P < .01, n = 4). An increase in TRH concentration was also found in the cerebrospinal fluid of SHR (30 +/- 3 versus 21 +/- 2 pg/mL, P < .01, n = 5), suggesting increased TRH release in the central nervous system. Northern blot analysis indicated a threefold augmented abundance of TRH precursor mRNA in the preoptic area of SHR. A polyclonal antibody raised against TRH injected peripherally or intracerebroventricularly lowered arterial blood pressure in SHR but not in WKY. In addition, long-term treatment with enalapril (5 mg/kg twice daily), which was effective in inhibiting serum angiotensin-converting enzyme activity by more than 50%, decreased arterial blood pressure and preoptic area TRH content of SHR, whereas another vasodilator, diltiazem (10 mg/kg every 8 hours), failed to produce a similar change.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7498978 TI - Changes of renal sympathetic activity in acute and chronic conscious sinoaortic denervated rats. AB - The arterial pressure level attained in sinoaortic denervated rats depends on the net effect of eliminating excitatory and inhibitory influences (chemoreceptor and baroreceptor elimination, respectively). After sinoaortic denervation is completed, the hypertension usually observed within the first few days is followed by normotension at the chronic stages. In this work renal sympathetic nerve activity was measured in conscious, unrestrained rats 6 hours (acute) and 20 days (chronic) after sinoaortic denervation. Increased arterial pressure (154 +/- 10 versus 114 +/- 3 mm Hg in controls) and renal sympathetic nerve activity (32 +/- 5 versus 13 +/- 2 bars per cycle in controls) with no changes in heart rate (404 +/- 17 vs 380 +/- 26 beats per minute) were observed in rats with acute sinoaortic denervation. In rats with chronic sinoaortic denervation, arterial pressure (119 +/- 8 mm Hg) and renal sympathetic nerve activity (13 +/- 6 bars per cycle) returned to control levels. Bradycardiac and tachycardiac responses to changes in blood pressure were reduced to 88% and 89%, respectively, in rats with acute sinoaortic denervation and 76% and 74%, respectively, in rats with chronic sinoaortic denervation. The reflex control of renal sympathetic nerve activity after acute and chronic sinoaortic denervation showed an impairment of sympathoinhibition (0.13 +/- 0.02 and 0.25 +/- 0.1 bars per cycle, respectively, versus 0.9 +/- 0.17 bars per cycle in controls). Sympathoexcitatory responses also were impaired in rats with acute and chronic sinoaortic denervation (0.08 +/ 0.03 and 0.37 +/- 0.1 bars per cycle, respectively, compared with 0.98 +/- 0.2 bars per cycle in controls).(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7498979 TI - Role of the rostral ventrolateral medulla in maintenance of blood pressure in rats with Goldblatt hypertension. AB - The aim of the present study was to examine the participation of the rostral ventrolateral medulla (RVLM) in the maintenance of hypertension in rats submitted to the renovascular Goldblatt (two-kidney, one clip) procedure. We inhibited or stimulated this area with the use of drugs such as glycine, L-glutamate, or kynurenic acid. (1) Bilateral microinjection of glycine (100 nmol, 200 nL, n = 13) into the RVLM of hypertensive rats produced a decrease in mean arterial blood pressure (MAP) from 177.2 +/- 29.3 to 102.3 +/- 20.9 mm Hg (P < .05), which was similar to the decrease produced by intravenous administration of hexamethonium. The inhibition of RVLM with glycine in normotensive rats produced a decrease in MAP from 106 +/- 17.1 to 59.7 +/- 7.3 mm Hg (P < .05, n = 9). (2) An impressive increase in MAP from 153.3 +/- 16.3 to 228 +/- 34.9 mm Hg (P < .05) occurred in hypertensive rats after microinjection of L-glutamate (50 nmol, 200 nL, n = 6) into the RVLM. The same procedure caused a significant but less intense increase in MAP from 105 +/- 13.8 to 148.3 +/- 24.9 mm Hg in normotensive rats (P < .05, n = 6). (3) A decrease in MAP from 151.6 +/- 25.3 to 96.8 +/- 22.5 mm Hg occurred in hypertensive rats after microinjection of the broad-spectrum glutamate antagonist kynurenic acid (4 nmol, 200 nL, n = 6) into the RVLM, whereas the same procedure did not change MAP in normotensive animals (n = 6). Heart rate was not significantly affected in any group.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7498980 TI - Noninvasive ambulatory 24-hour blood pressure in patients with high normal blood pressure and exaggerated systolic pressure response to exercise. AB - Few studies have investigated the significance of abnormal increases in systolic pressure during exercise in patients with high normal blood pressure and its correlation with 24-hour ambulatory blood pressure monitoring and left ventricular structure. This study was performed in 30 sedentary subjects (42 +/- 4 years old) with high normal blood pressure. Fifteen subjects presenting < 220 mm Hg systolic pressure during ergometric exercise were compared with 15 others with systolic pressure > or = 220 mm Hg. Average 24-hour (systolic, 127 +/- 5 versus 142 +/- 4 mm Hg, P < .01; diastolic, 82 +/- 4 versus 92 +/- 3 mm Hg, P < .01), daytime (systolic, 130 +/- 6 versus 144 +/- 4 mm Hg, P < .01; diastolic, 84 +/- 4 versus 92 +/- 4 mm Hg, P < .01), and nighttime (systolic, 116 +/- 7 versus 132 +/- 6 mm Hg, P < .01; diastolic, 72 +/- 6 versus 85 +/- 6 mm Hg, P < .01) ambulatory blood pressure monitoring values were significantly higher in subjects with an exaggerated blood pressure response to exercise. No significant differences were observed in left ventricular morphology. These findings indicate that subjects presenting high normal blood pressure and exaggerated systolic pressure during exercise show significantly high ambulatory blood pressure monitoring values that are not associated with left ventricular hypertrophy. PMID- 7498981 TI - Role of periaqueductal gray matter in hypertension in spontaneously hypertensive rats. AB - We performed experiments to study the effects of electrolytic lesions of periaqueductal gray matter on mean blood pressure, heart rate, and cardiac baroreflex in adult male spontaneously hypertensive rats. Cardiac baroreflex was assessed by the administration of randomly assigned doses of phenylephrine (0.3 to 5.0 micrograms/kg i.v.) or sodium nitroprusside (1.5 to 5.0 micrograms/kg i.v.) to unanesthetized rats. Bilateral lesions of the periaqueductal gray matter (0.5 mA/5 s) were then performed with rats under sodium pentobarbital anesthesia (35 mg/kg i.p.). Twenty hours after lesion, cardiac baroreflex was retested. Baroreflex data were analyzed by sigmoidal curve fitting. Lesion rats (n = 12) showed a significant decrease in both the gain (delta = -0.89 +/- 0.38 beats per minute [bpm]/mm Hg, P < .05) and curve midpoint (delta = -15 +/- 6 mm Hg, P < .05) of the cardiac baroreflex. Moreover, despite a moderate increase in heart rate (delta = 34 +/- 10 bpm, P < .01), resting mean blood pressure was significantly decreased 24 hours after the lesions (delta = -19 +/- 5 mm Hg, P < 01). No significant changes in cardiac baroreflex were observed in sham-lesion rats (n = 12). Histological examination showed circumscribed bilateral damage of dorsolateral periaqueductal gray matter. Dorsolateral periaqueductal gray matter is an area of the brain putatively related to fear and anxiety. It also projects onto premotor sympathetic neurons in the medulla. Although electrolytic lesions damage neurons as well as fibers of passage, these data suggest that dorsolateral periaqueductal gray matter has a far greater influence on resting cardiovascular control in spontaneously hypertensive rats than was previously suspected. PMID- 7498982 TI - Low-intensity exercise training attenuates cardiac beta-adrenergic tone during exercise in spontaneously hypertensive rats. AB - Acute and chronic exercise decrease peripheral sympathetic nerve activity, but the effect of exercise training of varying intensity on the sympathetic control of heart rate of spontaneously hypertensive rats has not yet been described. The effect of low and high intensities of exercise training on the vagal and sympathetic activities that control heart rate at rest and during dynamic exercise at 0.5, 0.8, and 1.0 mph for 4 minutes per stage was investigated in sedentary (SED, n = 11), high-intensity (HT, n = 12), and low-intensity exercise trained (LT, n = 13) spontaneously hypertensive rats. Exercise training was performed on a treadmill for 60 minutes, 5 days per week for 18 weeks, at 55% maximum oxygen consumption for the LT group and 85% for the HT group. Vagal and sympathetic activities were studied after administration of methylatropine (3 mg/kg) and propranolol (4 mg/kg), respectively. The LT group had a significantly lower heart rate (at 0.5, 0.8, 1.0 mph versus rest: 410 +/- 7, 464 +/- 9, and 295 +/- 6 beats per minute [bpm], respectively) than the HT (440 +/- 6, 453 +/- 7, 474 +/- 5, and 315 +/- 4 bpm) and the SED (474 +/- 11, 500 +/- 11, 523 +/- 10, and 327 +/- 3 bpm) groups. Sympathetic effect (LT: 84 +/- 10, 88 +/- 12, 105 +/- 12, and 9 +/- 4; HT: 123 +/- 8, 125 +/- 7, 133 +/- 7, and 34 +/- 7; SED: 130 +/- 13, 143 +/- 12, 150 +/- 10, and 38 +/- 7 bpm) and sympathetic tonus (LT: 125 +/- 6, 121 +/- 5, 112 +/- 6, and 91 +/- 6; HT: 145 +/- 9, 136 +/- 6, 142 +/- 8, and 118 +/- 7; SED: 136 +/- 6, 129 +/- 6, 132 +/- 7, and 118 +/- 8 bpm) were significantly decreased by low-intensity exercise training. In conclusion, low- but not high-intensity exercise training causes resting bradycardia and attenuation of tachycardiac response during progressive dynamic exercise in spontaneously hypertensive rats. This effect can be attributed to a significantly decreased beta-adrenergic tone that controls heart rate. PMID- 7498983 TI - Distinct mechanisms of upregulation of type 1A angiotensin II receptor gene expression in kidney and adrenal gland. AB - We previously demonstrated that type 1A angiotensin II (Ang II) receptor (AT1A) is the predominant renal subtype and is upregulated by a low sodium diet. We have now tested the hypothesis that upregulation of AT1A mRNA induced by sodium deficiency is renal specific and is mediated by activation of type 1 Ang II receptor (AT1). Male Wistar rats were divided into four groups (n = 5 each) and treated for 2 weeks with normal sodium diet (0.5%), normal sodium plus 3 mg/kg per day losartan, low sodium diet (0.07%), or low sodium diet plus losartan. At the end of the 2 weeks, body weight and mean arterial pressure were not different among the four groups (P > .05). Plasma renin activity was elevated by losartan treatment, sodium restriction, or the combination of the two versus control (P < .05). Northern blot analysis showed that the ratio of renal AT1A to glyceraldehyde 3-phosphate dehydrogenase (GAPDH) mRNA was increased by losartan treatment, sodium restriction, or the combination of the two versus control (P < .05). In contrast, the ratio of adrenal AT1A to GAPDH mRNA was increased only by sodium restriction versus three other groups (P < .05). Thus, sodium deficiency increases AT1A mRNA in both kidney and adrenal gland, while Ang II receptor blockade by losartan prevents low sodium-induced AT1A mRNA only in adrenal gland.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7498984 TI - Pressor and reflex sensitivity is altered in spontaneously hypertensive rats treated with angiotensin-(1-7). AB - We have suggested that angiotensin-(1-7) [Ang-(1-7)] may oppose the pressor activity of angiotensin II (Ang II). This hypothesis was supported by the fact that long-term intravenous infusion of Ang-(1-7) transiently lowers blood pressure in spontaneously hypertensive rats (SHR). We now investigated whether the pressor sensitivity to bolus injections of either phenylephrine (PE) or Ang II was altered on day 12 of an Ang-(1-7) infusion when blood pressure in the SHR had returned to hypertensive levels. SHR (n = 10) and WKY rats (n = 8) were given Ang-(1-7) intravenously via osmotic minipumps at a dose of 24 micrograms/kg per hour for 2 weeks. On day 12 of the infusion, mean arterial pressure and heart rate in halothane-anesthetized rats were similar in Ang-(1-7)-treated SHR (142 +/ 6 mm Hg; 388 +/- 9 beats per minute) and those infused with vehicle (146 +/- 5 mm Hg; 392 +/- 13 beats per minute). Pressor responsiveness to PE in Ang-(1-7) treated SHR was 22% less at a dose of 10 micrograms, while pressor responses to Ang II decreased by 20% and 25% at doses of 0.05 and 0.1 micrograms, respectively, compared with the vehicle-treated SHR (P < .05). There were no effects of the Ang-(1-7) infusion on pressor responses to Ang II or PE in WKY rats.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7498985 TI - Calcium channel blockers as inhibitors of angiotensin I-converting enzyme. AB - Using ion-exchange chromatography of dialyzed human urine from healthy and hypertensive patients, we detected two peaks of angiotensin I-converting enzyme (ACE) activity on hippuryl-His-Leu eluted at ionic strengths of 0.7 (F1 peak) and 1.25 (F2 peak) mS. These hydrolytic activities decreased gradually in the urine of patients submitted to isradipine treatment, F2 and F1 disappearing after 12 and 24 hours, respectively. By Western blot analysis, the urine fractions corresponding to both peaks from healthy and untreated patients presenting ACE activity and from treated patients (24 hours) without this activity were recognized by an ACE-specific antibody. These results indicated that ACE was present but inhibited in the urine of isradipine-treated patients. In vitro assays with ACE isolated from human urine and guinea pig plasma demonstrated that the enzyme is inhibited by isradipine and other commercially available calcium channel blockers, such as felodipine, nifedipine, and verapamil. A noncompetitive inhibition was observed with all calcium channel blockers studied. In conclusion, these results suggest that besides the primary effect on calcium channels, the more commonly used calcium channel blockers are also ACE inhibitors. The development of efficient calcium channel blockers with higher ACE inhibitory activity could result in interesting bifunctional antihypertensive drugs. PMID- 7498986 TI - Basal and angiotensin II-induced cytosolic free calcium in adult rat cardiomyocytes and fibroblasts after volume overload. AB - This study investigates basal and angiotensin II (Ang II)-induced [Ca2+]i concentrations in cells from hearts of rats that have undergone cardiac hypertrophy due to volume overload. [Ca2+]i measurements assessed by digital imaging using fura 2 methodology were performed on isolated ventricular cardiomyocytes and fibroblasts from adult rat hearts with a 4-week aortocaval shunt. Long-term aortocaval shunt induced a significant increase in atrial (72%) and ventricular (41%) weights and a large elevation in plasma atrial natriuretic peptide-(1-98) concentration (160%). For adult cardiomyocytes [Ca2+]i measurements are reported as diastolic (average of the lowest points) and systolic intracellular Ca2+ values (average of the maximum points corresponding to the diastolic points) over a 30-second time interval. Basal diastolic [Ca2+]i (99 +/- 4.1 nmol/L for experimental cells versus 90 +/- 4.8 for control cells) was not altered, whereas basal systolic [Ca2+]i was significantly greater in ventricular cardiomyocytes from overload hearts (155 +/- 2.3 versus 129 +/- 4.4 nmol/L for control cells, P < .05). Ang II increased intracellular Ca2+ spike frequency in a concentration-dependent manner in cardiomyocytes from control and overload myocardium. Basal and Ang II-induced intracellular Ca2+ spike frequencies were not modified in cardiomyocytes from hypertrophied hearts. Basal [Ca2+]i in ventricular fibroblasts from overload myocardium was significantly increased (128 +/- 5.1 nmol/L for fibroblasts from hypertrophied hearts versus 104 +/- 3.5 for control cells, P < .05). Ang II-induced [Ca2+]i was lower in fibroblasts from overload myocardium (P < .05).(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7498987 TI - Angiotensin-(1-7) potentiates the hypotensive effect of bradykinin in conscious rats. AB - Treatment with angiotensin-converting enzyme inhibitors increases the angiotensin (1-7) [Ang-(1-7)] and bradykinin concentrations in plasma and tissue. In this study we evaluated the interaction between these peptides by determining the effect of Ang-(1-7) on the hypotensive action of bradykinin in conscious rats. Administration of Ang-(1-7) (5 nmol) did not change mean arterial pressure or heart rate. However, the hypotensive effect of bradykinin, produced by an intravenous or intra-arterial route, was potentiated by Ang-(1-7) in a dose dependent manner. The Ang-(1-7) doses necessary to transform the effect of a single dose of bradykinin into that produced by a double dose (potentiating unit) were 2 nmol i.v. and 5 nmol IA. The Ang-(1-7) dose used did not change either the pressor effect of Ang II or the hypotensive effect of sodium nitroprusside. The bradykinin-potentiating Ang-(1-7) activity was significantly attenuated by pretreatment with indomethacin (5 mg/kg IM, n = 4). In an additional group the bradykinin-potentiating activity of Ang-(1-7) was evaluated 30 minutes after treatment with the angiotensin-converting enzyme inhibitor enalaprilat (10 mg/kg i.v., n = 9). Under this condition the bradykinin-potentiating activity of Ang-(1 7) was substantially increased, resulting in a potentiating unit of approximately 0.2 nmol IV. Pretreatment with indomethacin (5 mg/kg IM, n = 7) also attenuated the bradykinin-potentiating activity of Ang-(1-7) in enalaprilat-treated rats. These results show that Ang-(1-7) is a bradykinin-potentiating peptide in vivo. Furthermore, the data obtained with indomethacin suggest that prostaglandins participate in the mechanism of the bradykinin potentiation by Ang-(1-7). More importantly, these data suggest that the interaction between Ang-(1-7) and bradykinin can contribute to the pharmacological effects of angiotensin converting enzyme inhibitors. PMID- 7498989 TI - Inhibition of atrial natriuretic peptide excretory action by bradykinin. AB - We examined whether the excretory effect of atrial natriuretic peptide could be antagonized by intravenously administered bradykinin or by elevated endogenous kinin levels attained during converting enzyme inhibition. Urinary volume and sodium and potassium excretion were determined every 20 minutes in female, anesthetized Sprague-Dawley rats (weight, 0.19 to 0.22 kg) infused with 10 microL/min isotonic glucose. In some experiments, urinary cGMP content was measured by radioimmunoassay. Two intravenous boluses of 209 pmol (0.5 micrograms) atrial natriuretic peptide were given before and after the injection of test substances, and the response ratio was used to quantify inhibition. Single injections of 94.3 or 142 pmol (100 or 150 ng) bradykinin, 3 minutes prior to atrial natriuretic peptide, inhibited the excretion of water, sodium, and potassium by 70%, 75%, and 50%, respectively. Larger (236 to 472 pmol) or smaller (23.6 to 47.2 pmol) bradykinin doses were ineffective. None of the bradykinin doses tested affected basal urinary output, systemic pressure, or the modest depressor effect of atrial natriuretic peptide. The anti-atrial natriuretic peptide effect of bradykinin was completely prevented by the kinin receptor antagonist Hoe 140. Converting enzyme inhibition with ramipril (96 nmol IV) also blunted atrial natriuretic peptide diuresis and natriuresis by 70% and reduced urinary cGMP excretion by 50%. These effects of ramipril were mediated by endogenous kinin accumulation, since they were abolished by pretreatment with Hoe 140. It is concluded that intrarenal kinins modulate the renal actions of atrial natriuretic peptide, and at a precise concentration bradykinin strongly antagonizes atrial natriuretic peptide by preventing its transduction mechanism. PMID- 7498988 TI - Comparison of candoxatril and atrial natriuretic factor in healthy men. Effects on hemodynamics, sympathetic activity, heart rate variability, and endothelin. AB - The purpose of these experiments was to compare the effects of endopeptidase inhibition with oral candoxatril on systemic and forearm hemodynamics and muscle sympathetic nerve activity with responses to a low-dose atrial natriuretic factor infusion. Eleven healthy men received at random on three separate days either intravenous saline, natriuretic factor (1.6 pmol/kg per minute) plus saline, or oral candoxatril (200 mg) plus saline. Measurements were made at baseline and 30, 60, and 90 minutes after interventions. Atrial natriuretic factor lowered diastolic pressure (P < .01), central venous pressure (P < .001), forearm blood flow (P < .05), and forearm vascular compliance (P < .05) but had no effect on systolic pressure, heart rate or its variability, stroke volume, sympathetic nerve activity, plasma norepinephrine, or endothelin-1. Plasma epinephrine increased (P < .01). Candoxatril lowered central venous pressure (P < .001) and increased systolic pressure (from 116 +/- 6 to 120 +/- 7 mm Hg; P < .05), endothelin (from 4.6 +/- 1.1 to 6.8 +/- 3.2 pmol/L; P < .02), and epinephrine (P < .05), without affecting any other variables. Candoxatril and atrial natriuretic factor lowered central venous pressure in healthy men without causing a reflex increase in sympathetic nerve activity or norepinephrine, yet epinephrine rose. This suggests that both interventions may specifically inhibit sympathetic nerve traffic to muscle at physiological plasma atrial natriuretic factor concentrations. However, whereas the peptide lowered blood pressure, candoxatril increased systolic pressure. These contrasting hemodynamic responses may be related to differences in plasma atrial natriuretic peptide concentration and to altered endothelin metabolism by candoxatril. PMID- 7498990 TI - Stress-induced elevation of ouabainlike compound in rat plasma and adrenal. AB - Recent observations demonstrate the presence of neurosteroids and their rapid increase in response to acute stress. In view of a steroidal nature of ouabainlike compound, we tested the hypothesis that ouabainlike compound may participate in a homeostatic response to acute stress. Male Wistar rats were subjected to acute stress by swimming in water (22 degrees C) for 10 minutes. The levels of ouabainlike compound in plasma, hypothalamus, pituitary, and adrenal at 10, 40, and 70 minutes (n = 8 for each) after the end of swim stress were compared with nonstressed control levels (n = 10). Ouabainlike compound was measured by a radioimmunoassay for ouabain. Plasma levels of corticosterone and catecholamines were also measured. Plasma corticosterone concentrations increased rapidly at 10 minutes (P < .01) and then declined. A trend for a rise in plasma catecholamines was found at 10 minutes. Adrenal levels of ouabainlike compound concomitantly increased at 10 minutes (P < .01, control: 58.9 +/- 5.9 pmol ouabain equivalents per gram; 10 minutes: 92.5 +/- 4.8; 40 minutes: 47.3 +/- 9.6; 70 minutes: 45.1 +/- 6.3). In contrast, the response of plasma ouabainlike compound was slow and doubled at 40 minutes (P < .01, control: 115 +/- 12 pmol ouabain equivalents per liter; 10 minutes: 132 +/- 23; 40 minutes: 226 +/- 53; 70 minutes: 117 +/- 16). Ouabainlike compound levels in hypothalamus and pituitary remained unaltered. These findings suggest that ouabainlike compound may function as a stress hormone. PMID- 7498991 TI - Regulation of neuronal calcitonin gene-related peptide expression. Role of increased blood pressure. AB - Calcitonin gene-related peptide (CGRP) is a potent vasodilator neuropeptide. We have previously demonstrated that CGRP mRNA levels are increased in dorsal root ganglia, and immunoreactive CGRP content is elevated in the spinal cord in mineralocorticoid-salt hypertension. Dorsal root ganglia neuronal cell bodies synthesize CGRP and send axons peripherally to blood vessels and centrally to spinal cord sites involved in blood pressure regulation. This increased synthesis of a potent vasodilator is a compensatory response to attenuate the increase in blood pressure; however, it is not known if neuronal CGRP is regulated simply by the elevated blood pressure or by changes in other parameters. To determine if elevation of blood pressure in normal rats induced by the administration of a potent vasoconstrictor can increase neuronal CGRP mRNA, 7-week-old male Sprague Dawley rats were treated for 2 weeks with either angiotensin II (n = 6) or vehicle (n = 6) by using implanted osmotic minipumps. After the treatment period, the angiotensin II-treated rats displayed a marked increase in systolic blood pressure (angiotensin II, 217 +/- 18 versus control, 131 +/- 3 mm Hg, P < .001), and decrease in plasma renin activity (angiotensin II, 3.7 +/- 3.5 versus control, 35.9 +/- 14.2 ng.mL-1.h-1, P < .05). However, dorsal root ganglia CGRP mRNA content did not significantly differ between the two groups of rats. These results demonstrate that a marked increase in blood pressure, by itself, does not increase neuronal CGRP mRNA accumulation.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7498992 TI - Adenosine A1 receptor mRNA in microdissected rat nephron segments. AB - Adenosine plays several roles in the kidney mediated by the specific receptors A1, A2, and possibly A3. We studied the localization of adenosine A1 receptor mRNA in rat nephron segments using reverse transcription and polymerase chain reaction (RT-PCR). The nephron segments of male Sprague-Dawley rats (6 to 8 weeks old) were microdissected. Total RNA was prepared by the acid-guanidinium-phenol chloroform method and used in the following RT-PCR assay. Because the PCR primers spanned no intron, samples reacted in the absence of RT were used as controls for amplification of genomic DNA. The PCR products were size-fractionated by electrophoresis, visualized with ethidium bromide staining, and confirmed by Southern blot analysis. PCR products were detected in all of the nephron segments examined. No signals were detected in samples reacted in the absence of RT. Strong signals were detected in glomeruli, medullary collecting duct, cortical thick ascending limb, and medullary thick ascending limb, while weak signals were found in proximal convoluted and straight tubules. Previously, the presence of A1 receptors has been demonstrated in glomeruli, collecting duct, and thick ascending limb in the rat kidney by autoradiography and binding studies. In addition to these segments, we further detected A1 receptor mRNA in proximal convoluted and straight tubules. Thus, A1 receptor mRNA seems to be broadly expressed along the nephron. PMID- 7498993 TI - Cardiovascular effects of a specific nonpeptide antagonist of substance P (NK-1) receptor in DOCA-salt hypertension. AB - The neurotransmitter substance P acts also as a potent vasodilator. Its participation in the pathogenesis of deoxycorticosterone acetate (DOCA)-salt hypertension was evaluated by an acute infusion of a newly synthesized, potent, specific nonpeptide antagonist of substance P at the NK-1 receptor, the agent CP 96,345. In conscious unrestrained rats, CP 96,345 induced significant and sustained increases in mean arterial pressure of DOCA-salt rats but only small, transient, and nonsignificant rises in blood pressure of sham-treated control rats. The rise in blood pressure was not accompanied by changes in heart rate. Maximal blood pressure increase in DOCA-salt rats was 31.7 +/- 14.8 mm Hg. In a second series of experiments, the hemodynamic effects of this antagonist were evaluated under anesthesia in both DOCA-salt and sham-treated control rats by the thermodilution method. During CP 96,345 infusion, sustained increases in cardiac index and stroke volume and decreases in total peripheral resistance were observed in both DOCA-salt and control rats. In DOCA-salt rats, cardiac index rose by 79.4%, while total peripheral resistance fell by 27.9% of the baseline values. In control rats, the changes were smaller (+27.2% and -22.5%, respectively). Stroke volume changed in parallel to cardiac output in both groups. The data suggest that acute blockade of NK-1 receptors increases blood pressure in DOCA-salt rats mainly by an increase in cardiac output. We conclude that endogenous substance P tends to counteract the DOCA-salt-induced elevation of blood pressure by modulating both cardiac output and peripheral resistance. PMID- 7498994 TI - Role of anthropometric indexes and blood pressure as determinants of left ventricular mass and geometry in adolescents. The Rio de Janeiro Study. AB - To evaluate left ventricular structural changes and their relationship to blood pressure and anthropometric indexes, we examined by echocardiography 108 adolescents aged 13 to 19 years. Subjects were divided into three groups according to blood pressure tracking during three moments of observation: group 1 (n = 27), > or = 95th percentile; group 2 (n = 37), < or = 50th percentile; and group 3 (n = 44), blood pressure not stable in the original percentile. Left ventricular mass index and the prevalence of altered left ventricular geometry were greater in group 1 (P < .05 and P < .02, respectively). Of all the anthropometric indexes, body surface area showed the best correlation with left ventricular mass (P < .00001). Left ventricular mass also correlated with systolic and diastolic pressures (P < .00001 and P < .003, respectively). Ventricular septal and posterior wall thicknesses and left ventricular diastolic diameter showed good correlations with body surface area (P < .00001). These variables also correlated with systolic pressure (P < .001). In a multiple regression model when body surface area was controlled, systolic pressure did not correlate significantly with left ventricular mass. In a similar model systolic pressure maintained a significant correlation with ventricular septal and posterior wall (P < .00001) thicknesses but not with left ventricular diastolic diameter (P > .05). We conclude that left ventricular structural changes can occur early after initial abnormalities of blood pressure. Considering that body surface area and systolic pressure were the best predictors of left ventricular alterations in adolescents, the usual way of correcting left ventricular mass by body surface area should be reviewed. PMID- 7498995 TI - Relationship between systemic hemodynamics and ambulatory blood pressure level are sex dependent. AB - Sex-related differences in systemic hemodynamics were analyzed by means of cardiac index and systemic vascular resistance according to the level of daytime ambulatory blood pressure. In addition, we assessed the relations between ambulatory blood pressure measurements and systemic hemodynamics in male and female patients. We prospectively included 52 women and 53 men referred to our unit for evaluation of arterial hypertension. Women and men were grouped according to the level of daytime mean arterial pressure: < 110 or > or = 110 mm Hg. Patients underwent noninvasive evaluation of resting hemodynamics (impedance cardiography) and 24-hour ambulatory blood pressure monitoring. Compared with women men with lower daytime blood pressure had a 12% higher systemic vascular resistance index (P = NS) and a 14% lower cardiac index (P < .02), whereas men with higher daytime blood pressure had a 25% higher vascular resistance (P < .003) and a 21% lower cardiac index (P < .0004). Furthermore, in men systemic vascular resistance correlated positively with both daytime and nighttime systolic and diastolic blood pressures, whereas cardiac index correlated negatively only with daytime diastolic blood pressure. In contrast, women did not exhibit any significant correlation between hemodynamic parameters and ambulatory blood pressure measurements. In conclusion, sex-related differences in systemic hemodynamics were more pronounced in the group with higher daytime hypertension. The relations between systemic hemodynamics and ambulatory blood pressure level depended on the sex of the patient. In men a progressive circulatory impairment underlies the increasing level of ambulatory blood pressure, but this was not observed in women. PMID- 7498996 TI - Blood pressure influences the occurrence of complex ventricular arrhythmia in hemodialysis patients. AB - We investigated the relationship between blood pressure and the occurrence of complex ventricular arrhythmias (multiform, couplets, or runs) as assessed by 48 hour Holter monitoring in 74 stable long-term hemodialysis patients (44.5 +/- 12 years old; 54% men; 74% whites; dialysis duration, 51.3 +/- 36.1 months; systolic pressure, 146.6 +/- 19.3 mm Hg; diastolic pressure, 89.2 +/- 12.1 mm Hg; prevalence of arterial hypertension, 33.8%). Systolic and diastolic pressures represented the average of all predialysis determinations during the 3 months preceding the tests. Hemodialysis was performed midway through the Holter monitoring period. M-mode and bidimensional echocardiograms and myocardial perfusion tests were also obtained from all patients. Complex arrhythmias were observed in 37 individuals (50%). Univariate analysis showed that systolic pressure (P < .001), diastolic pressure (P < .05), age (P < .001), left ventricular posterior wall thickness (P < .01), left ventricular mass index (P < .05), and ischemic alterations on myocardial perfusion tests (P < .005) were significantly associated with complex arrhythmias. With the use of a multivariate model (stepwise logistic regression analysis) only systolic pressure (P < .01) and age (P < .05) were independently associated with complex arrhythmias. Sex; angina; dialysis duration; New York Heart Association functional class; use of digitalis; plasma levels of creatinine, sodium, potassium, calcium, and phosphate; hematocrit; left ventricular fractional shortening; left ventricular diastolic diameter; and ST segment deviation were not correlated with complex arrhythmias. The severity and frequency of complex arrhythmias were not influenced by hemodialysis. At follow-up (5 to 80 months) 5 patients had died of sudden death, 4 of whom were hypertensive and older than 45 years.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7498997 TI - Evaluation of the technique used by health-care workers for taking blood pressure. AB - The precise guidelines recommended by the American Heart Association for blood pressure measurement are commonly overlooked by health-care workers, who generally take blood pressure in an arbitrary way. To validate this observation we designed a descriptive and observational study to be carried out in a major hospital. One hundred and seventy-two health-care workers divided into four groups (63 general practitioners, 25 clinical and 25 surgical specialists, and 59 nurses) were evaluated in a two-part test. In the first part (practical), the examinee had to follow all the steps recommended by the American Heart Association to get a passing score. In the second part (theoretical, which came second to avoid influencing the practical), the examinee had to answer correctly 7 of 10 questions based on the American Heart Association's guidelines to obtain a passing score. The highest accepted variation in systolic and diastolic pressures between examinee and observer was +/- 4 mm Hg. None of the examinees followed the American Heart Association's recommendations. Sixty-three percent of systolic and 53% of diastolic readings were out of range. Surgical specialists obtained the best practical results (48% systolic and 64% diastolic within range), and nurses obtained the lowest values (29% and 39%, respectively; P = .03 versus surgical specialists). These two groups showed deficiencies in the theoretical test (nurses, 10% correct answers and surgical specialists, 16%). Clinical specialists obtained the best results on the theoretical test (60% correct; P < .05 versus the other groups) but were deficient in the practical test (32% systolic and 60% diastolic within range).(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7498998 TI - Malignant hypertension is accompanied by marked alterations in chylomicron metabolism. AB - Malignant hypertension (MH) is a severe complication of untreated arterial hypertension that damages the vascular system. It is often accompanied by disturbances in lipid metabolism that could contribute to its pathophysiology. We examined chylomicron metabolism in MH patients using a triglyceride-rich emulsion known to mimic natural chylomicrons when injected into the bloodstream. The emulsion was labeled with [3H]triolein and [14C]cholesteryl oleate and injected intravenously into 15 normolipidemic MH patients aged 29 to 56 years (8 men) for comparison with 17 healthy control subjects. Consecutive plasma samples were taken at regular intervals during 1 hour for determination of the disappearance curves of the labels. The fractional clearance rate of the [3H]triolein emulsion in MH patients was twice as small as that of control subjects (0.061 +/- 0.012 and 0.141 +/- 0.074 min-1, respectively). On the other hand, [14C]cholesteryl oleate fractional clearance rate was not statistically different in MH patients and control subjects (0.032 +/- 0.004 and 0.056 +/- 0.014 min-1, respectively). These results indicate that in MH, lipolysis (measured by the fractional clearance rate of [3H]triolein) is pronounced diminished, whereas the removal of the remnant particles (measured by the fractional clearance rate of [14C]cholesteryl oleate) is not importantly affected. In conclusion, there is an alteration in the circulatory transport of dietary lipids that may be an important component in the vascular disease associated with MH. PMID- 7498999 TI - Selective decontamination of the digestive tract: indications and problems. PMID- 7499000 TI - Genotypes and virus load in patients with hepatitis C infection. AB - Virus load was tested by means of PCR calibrated with standards and HCV genotypes were determined by the LIPA-technique using sera from 123 HCV patients. Of these 39 were on renal hemodialysis treatment, 19 suffered from hemophilia, 13 were i.v. drug users and the remaining 52 had none of these risk factors (chronic hepatitis group). The most prevalent subtype in Austria was 1b followed by 3a and 1a. However, genotype 1b infections were found relatively less often in hemophilia patients and drug users than in the other groups, indicating that hemophiliacs probably had been infected by an antihemophilic plasma coming from South American or Asian donors. The highest amounts of virus were found in patients infected with genotype 3a. Determination of the patient's virus load and of the infecting subtype of HCV may be helpful in planning interferon alpha therapy. PMID- 7499001 TI - Symptoms and complications of pertussis in adults. AB - There is increasing evidence that pertussis occurs frequently in adults, but there is limited information on the clinical course of this disease beyond childhood. A household contact study on the efficacy of an acellular pertussis vaccine was used to study the symptoms of pertussis in adults. Among 257 patients with pertussis identified in 121 families during a two-year period in one study center with a low whole-cell pertussis-vaccine uptake, 79 (30.7%) were adults, aged 19-83 years (mean age: 36 years) with a 1:1.8 male to female ratio. Ninety one percent of the adults suffered from coughing (mean duration: 54 days), and in 80% this cough lasted > or = 21 days. Whoops were rare (8%), whereas cough followed by vomiting and/or choking (53%) and cough disturbing sleep (52%) were common. This is the first report to describe sweating attacks as symptom of pertussis (14%). Pharyngeal symptoms (37%), influenza-like symptoms (30%), sneezing attacks (22%), hoarseness (18%), sinus pain (16%) and headaches (14%) were also observed. Various complications were seen in 23% of the patients. In order to minimize the spread of the organism, microbiological diagnostics should be vigorously applied to all symptomatic contacts of a patient with pertussis but also to all patients with long lasting cough-irrespective of age. PMID- 7499002 TI - Pattern of soluble TNF receptors I and II in sepsis. AB - The serum levels of soluble TNF receptors I (sTNFR I) and sTNFR II were measured frequently in 14 patients with sepsis to evaluate the pattern of these TNF antagonists in relation to TNF alpha, Soluble TNFR I and II could be detected in all samples with significantly higher levels (p < 0.001) compared to healthy controls. The concentration of sTNFR I as well as sTNFR II was significantly higher in nonsurvivors compared to survivors during the first 36 h of sepsis (p < 0.001). Levels remained elevated throughout the evaluation with maximal values in patients who died. A positive correlation exists between both receptors and between soluble receptors and simultaneously obtained sepsis score (p < 0.01) while TNF immunoreactivity detected in 80% of all samples did not correlate to soluble receptor levels or sepsis score. Soluble receptors were constantly found in the circulation representing the inflammatory state throughout the evaluation even when TNF activity was undetectable. PMID- 7499003 TI - Cerebral tuberculoma--a comparative study in patients with and without HIV infection. AB - The microbiological, clinical and radiological findings of cerebral tuberculomas in four patients with and in five patients without HIV infection were compared. The study was carried out during the last 14 years. The CT scans were analyzed in a blinded fashion. Cerebral tuberculoma in HIV-negative patients was clinically characterized by seizures, while in HIV-positive patients this finding was absent. All four HIV-infected patients had headache and fever and their CSF showed lymphocytic meningitis. Two HIV-negative and three HIV-positive patients had concurrent extracerebral tuberculosis. In HIV-infected patients, the cerebral tuberculoma was a secondary finding of disseminated tuberculosis. In our small patient samples, the cerebral tuberculoma presented as spontaneous hypodense cerebral lesions in all the HIV-positive patients but as a hyperdense cerebral lesion in the HIV-negative patients. Two patients of each group had ring enhancement lesions. Cerebral tuberculoma was diagnosed in about 4 weeks for HIV positive patients, but took some 16 weeks for HIV-negative patients, the latter being first suspected of having a cerebral tumor or bacterial abscess. Diagnostic craniotomy was thus necessary for the HIV-negative patients. One patient of each group died as a consequence of cerebral tuberculoma, all the remaining patients improved with treatment. PMID- 7499004 TI - Synergistic interaction between ofloxacin and cefotaxime against common clinical pathogens. AB - Antimicrobial synergy resulting from combined antibiotic therapy is often important in the treatment of serious bacterial infections. To investigate the interactions between cefotaxime (CTX), desacetylcefotaxime (DES), and ofloxacin (OFL), 247 recent clinical isolates were tested for in vitro susceptibility to each antibiotic alone by an agar dilution technique and retested with the various antibiotic combinations using a checkerboard protocol. Fractional inhibitory concentrations were calculated for all organisms with all drug combinations. Time kill kinetic studies were performed on selected isolates to examine the bactericidal activity of the various antimicrobial combinations. Of the 110 gram negative organisms tested, synergy or partial synergy between CTX, DES and OFL was demonstrable for 89 (81%). Included in the study were 70 members of the Enterobacteriaceae family, 20 isolates of Pseudomonas aeruginosa, 10 strains of Acinetobacter baumannii, and 10 isolates of Xanthomonas maltophilia. Additive activity was observed against an additional 13 (11%) isolates. Findings were similar for the 89 gram-positive isolates examined. Organisms tested included methicillin-resistant Staphylococcus aureus (20), methicillin-susceptible Staphylococcus aureus (20), methicillin-resistant Staphylococcus epidermidis (9), methicillin-susceptible S. epidermidis (10), Enterococcus faecalis (10), and Streptococcus pneumoniae (20). Synergy or partial synergy was observed against 81 (91%). Less synergistic activity was detected, however, with members of the Bacteroides fragilis group. Of the 48 organisms tested, synergy or partial synergy was noted for only 27 (57%). Isolates representative of each major group of organisms included in the study were tested to determine whether synergistic bactericidal activity was also demonstrable with the three drugs. Time kill studies supported the checkerboard results.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7499005 TI - Klebsiella oxytoca meningitis: frequent association with neurosurgical procedures. AB - Klebsiella oxytoca meningitis is a rare condition. Nine patients were diagnosed between 1981 and 1993 at our institution. These accounted for 2.3% of 393 patients with blood and/or cerebrospinal fluid culture-proven bacterial meningitis. K. oxytoca was noted in both community-acquired meningitis and nosocomial meningitis. Eight of the nine cases were patients who had undergone neurosurgical procedures. Four were mixed bacterial meningitis. All K. oxytoca isolates were susceptible to third-generation cephalosporins and all but one to chloramphenicol. Antibiotic therapy was successful in eight patients but failed in one. PMID- 7499006 TI - Serum and cellular pharmacokinetics of clarithromycin 500 mg q.d. and 250 mg b.i.d. in volunteers. AB - In an open-label, randomized, crossover study 12 healthy volunteers were given clarithromycin orally 250 mg twice daily (b.i.d.) and 500 mg once a day (q.d.). Blood and saliva samples were collected on study days 1 and 5 to determine the pharmacokinetics of clarithromycin and its 14-hydroxy metabolite in plasma and saliva, and to measure concentrations of clarithromycin in mononuclear cells (MNCs) and polymorphonuclear leucocytes (PMNs). The mean peak levels of clarithromycin on day 5 of therapy in serum (2.3 vs. 1.2 mg/l), saliva (1.1 vs. 0.3 mg/l) and blood cells 60 vs. 26 mg/l in MNCs and 29 vs. 14 mg/l in PMNs) were at least doubled, the trough levels were lower with 500 mg q.d. vs. 250 mg b.i.d. (0.09 vs. 0.28 mg/l in serum; 0.06 vs. 0.13 mg/l in saliva; < 1 vs. 6.8 mg/l in MNCs; 0.8 vs. 2.8 mg/l in PMNs). The mean relative peak serum concentrations of the 14-hydroxy metabolite were somewhat lower with the 500 mg dosage (0.78 vs. 0.46 mg/l). The peak concentrations of clarithromycin and its 14-hydroxy metabolite in saliva were 25-40% and 50-80% of the maximum serum concentrations with both dosage regimens. Clarithromycin exhibits good and rapid penetration into intracellular as well as into extravasal extracellular body compartments. Clarithromycin 500 mg q.d. compares favourably with 250 mg b.i.d., as far as peak serum levels and bioavailability are concerned, but trough levels are lower at the end of the 24-hour dosing interval. PMID- 7499008 TI - Acute suppurative thyroiditis caused by Salmonella enteritidis. AB - Acute suppurative thyroiditis in a 47-year-old diabetic woman is reported. Diagnosis was suspected by clinical examination and confirmed by ultrasound, CT scan and cytological examination. A rare causative agent was isolated--Salmonella enteritidis. No signs of nodular goiter or connection with piriform sinus were found. In spite of our efforts, the source and route of infection remained unclear. However, the haematogenic route seems to be the most plausible. Surgical drainage and antibiotics brought about a complete recovery. Partial lesion of the gland required transitory administration of levothyroxine. PMID- 7499007 TI - Evaluation of the detection of Borrelia burgdorferi DNA in urine samples by polymerase chain reaction. AB - It is difficult in some cases to identify an infection caused by Borrelia burgdorferi and to monitor the effect of therapy. Seropositivity will persist even after successful treatment and therefore may suggest ongoing infection. For direct detection of B. burgdorferi DNA in human urine samples, the polymerase chain reaction (PCR) was evaluated. A published primer system was selected, which amplifies a 259 bp fragment from the gene encoding the 23S rRNA. The lower detection limit of the primer system was 10 fg of extracted B. burgdorferi DNA. Several methods for the pretreatment of urine samples were tested. Of these, the Geneclean kit (Bio 101, USA) showed the best results. A total of 114 urine samples from 74 patients belonging to three clinical groups was investigated: (i) 51 samples from 26 patients with active Lyme disease, (ii) 36 samples from 27 patients with previous infection but no symptoms at the time the urine was collected, and (iii) 27 samples from 21 seronegative control patients without Lyme disease. B. burgdorferi DNA was detected in 25 urine samples of 17 patients with active disease, whereas 26 samples from this group of patients were negative. Only one asymptomatic case with previous infection showed a positive result, and the urine samples of the patients without Lyme disease were uniformly negative. Two of four patients from whom samples before and directly after onset of therapy were available converted from negative to positive PCR results after initiation of therapy, accompanied by the symptoms of a Jarisch-Herxheimer reaction.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7499010 TI - Re.: Oral administration of a certain strain of live Escherichia coli for intestinal disorders? (Infection 23 [1995] 51-54) PMID- 7499009 TI - Visceral leishmaniasis during pregnancy treated with meglumine antimoniate. AB - Data on the efficacy and safety of pentavalent antimony in the treatment of visceral leishmaniasis during pregnancy are scanty. A case of visceral leishmaniasis in a 39-year-old woman in the second trimester of pregnancy is reported here. The patient was hospitalized in poor condition with high fever and pancytopenia which had lasted for 6 weeks. A bone marrow aspirate revealed numerous amastigotes and serodiagnosis for Leishmania was positive at a high titer. The patient was successfully treated with meglumine antimoniate at a daily dose of 850 mg of antimony for 20 days. She delivered at term a healthy female baby who remains in good condition at 18 months of age. Thus a dose of 850 mg of antimony, which is lower than that presently recommended, seems to be effective and non toxic to the fetus when administered at the second trimester of pregnancy. PMID- 7499011 TI - T-lymphocyte subpopulations as factors predicting clinical response to interferon in hepatitis C virus-related chronic active hepatitis. PMID- 7499012 TI - Real time ultrasound for venous catheter placement. PMID- 7499013 TI - Uncomplicated central vein catheterization of high risk patients with real time ultrasound guidance. AB - Internal jugular vein cannulation has become a routine and clinically important aspect of medical care of critically ill patients. The landmark guided technique usually affords rapid and easy vascular access, but is not always successful and may be complicated by arterial puncture, hematoma, or pneumothorax. Some categories of patients, in particular patients with no external landmarks and patients with coagulopathies, appear to be at an increased risk of complications. We report the experience of internal jugular vein cannulation by a single operator with the external landmark technique in 10 patients and with ultrasound guidance in 31 patients, including 12 high risk patients. These patients had severe coagulopathies due to hepatic failure, HELLP syndrome, excess of anticoagulation treatment, or they had no external anatomic landmarks because of anasarca or obesity, were unable to maintain the horizontal position, or were external landmark catheterization failures. With the availability of the ultrasound device, success and complication rates improved markedly, suggesting that the ultrasound technique is easy to learn and rapidly produces an improvement over the external landmark method. In particular the 13 cannulations performed in 12 high risk patients were all successful at the first attempt, with no complications. In the overall population successful cannulations improved from 80% to 100%, first attempt success from 20% to 87% and carotid punctures decreased from 33% to 3.2%. Our results confirm that ultrasound guided cannulation of the internal jugular vein allows safer operation in high risk patients or when access problems are anticipated. PMID- 7499015 TI - Intraoperative blood salvage (IOBS) in cardiac and vascular surgery. AB - The extensive application of IOBS has permitted a great reduction in the use of homoglous transfusion which presently represents the largest field of application of autologous systems. In cardiac and vascular surgery, IOBS is particularly useful to the goal of preventing the transmission of viral disorders and other adverse effects related to homologous transfusions. The apparatuses for IOBS may also be used to perform hemodilution and sequestration of a desired amount of platelet rich plasma. The appropriate usage of drugs in perioperative period and the promotion of hemostasis with IOBS are important costituents for the correct transfusional management of the patient. The feasibility and safety of IOBS is known and in expert hands it is an optimal method for the transfusional treatment of surgical patients. PMID- 7499014 TI - Urea, creatinine and phosphate kinetic modeling during dialysis: application to pediatric hemodialysis. AB - The kinetics of urea, creatinine and phosphate removal during dialysis were investigated in pediatric patients using a two-pool model taking into account fluid shifts and mass transfer between the two compartments. It is found that even urea must be described by a two-pool model since it presents a post dialysis rebound due to equilibration between the two compartments. Phosphate plasma concentration drops very sharply during the first hour of dialysis and rises rapidly during the rebound period. This pattern cannot be accounted for by the classical two-pool model with constant generation rate and mass transfer coefficients, but corresponds to a large time-dependent phosphate influx from the intracellular compartment in which phosphate is generated by biochemical reactions or liberated from the bones. This influx was calculated for four patients representing 8 dialysis sessions and was found to reach a plateau after 90 minutes of dialysis, dropping rapidly during the rebound period. PMID- 7499016 TI - Intraoperative blood salvage: a new artificial organ? AB - Optimal blood supply is critical to modern medical practice. Among the different possibilities of improving the quality and safety of blood, it is generally felt that autologous donation has played an important role and has contributed to changing transfusional practices, mainly since the appearance of HIV and HCV on the blood transfusion scene. At the San Martino Hospital Immunohematology Service, the autotransfusion era began in 1985. Autologous predeposit donation was the first to be introduced, followed by intentional perioperative hemodilution, intraoperative blood salvage with DFC apparatuses and lastly post operative blood salvage. From about 200 autologous donations in 1985 we reached 5,372 in 1993 and more than 6,000 autologous donations are expected for 1994. Only 189 intraoperative blood salvages, were carried out in 1986, 593 in 1989, 1,207 in 1993 and more than 1,500 blood salvage sessions are anticipated for 1994. In the meantime, the total number of homologous RBC units employed in the Hospital dropped from 45,000 in 1985 to 18,000 in 1994, with the Onco hematological Divisions using approximately 10,000 units of packed RBC. PMID- 7499017 TI - From low hematocrit physiology to isovolemic hemodilution. AB - The aim of this study, starting from some concepts of physiology, was to deal with the ever growing question of intentional anemia. Physiology suitably expresses the linear relationship between hematocrit and blood fluidity, and at the same time explains how viscosity and aggregability are important co-factors in the circulation, in disease and in thromboembolic complications. The need to understand how a low hematocrit level, compared to the volemia, is a fundamental requirement in surgery, and even more so in surgery with a high risk of embolism, like orthopedic traumatology surgery. Normovolemic hemodilution was analyzed both for the risks it involves and for how much it sticks to the changes induced by a similar situation on the various organs and apparatus. The authors, based on their experience in the field of anesthesiology, propose an operating schedule to safely perform intentional normovolemic hemodilution, describing the preoperative, intraoperative and finally the postoperative period. In conclusion, the authors advocate a wider use of this technique, confirming its safety in relation to volemia. PMID- 7499018 TI - Blood saving program: a multicenter Italian experience. AB - To assess the causes of failure in a blood saving program and the consequent need for homologous transfusions during major orthopedic surgery, data were collected from six orthopedic centers where an autotransfusion program was carried out from 1992-1994. The 1,576 (1,016 female and 560 male) patients studied (average age 63.6 +/- 10 years) were treated either for total hip prosthesis (1,144 pts) total knee prosthesis (212 pts), plate and screw removal plus hip prosthesis (8 pts), hip revision (cup 48 pts, stem 10 pts, cup plus stem 146 pts) and total knee revision (8 pts). Ninety percent of patients did not undergo homologous transfusions, which was significantly connected to: male sex (8% versus 11.7% p = 0.02), younger age (63.2 vs 66.9 years, p = 0.0001), higher weight (male 76 vs 70.7 Kg. p = 0.003, female 65.6 vs 61.9 Kg. p = 0.0003); higher baseline Hb (13.9 vs 12.8 g/dl, p = 0.0041), shorter operation time (110 vs 128 minutes, p = 0.0001); use of indobufen for antithromboembolic prophylaxis (indobufen 4.6% vs heparin calcium 15.2% vs low molecular weight heparin 9% p = 0.0001). On the contrary, the factors favoring homologous transfusions were: peripheral anesthesia (spinal and epidural 15.3% vs general 9.3% vs integrated 9.8% p = 0.016), total hip or knee revision, fixation device removal before hip prosthesis, systemic hypertension and coronary heart disease. PMID- 7499020 TI - Biopolymeric modification of superoxide dismutase (mPEG-SOD) to prevent muscular ischemia-reperfusion damage. AB - The efficacy of preventing ischemia-reperfusion damage by employing native or modified (mPEG-SOD) superoxide dismutase in an experimental animal model of acute ischemia of the left hindlimb was tested. Four hours and thirty minutes complete warm ischemia was induced in the left hindlimb of 43 Wistar rats, by clamping the femoral artery and monitoring its efficacy with Laser Doppler flowmetry. After ten days, a significative difference (p = 0.004) of the survival leg rate was found in the group treated with mPEG-SOD (86.6%) compared with the control group (30%). Histomorphological and ultrastructural analysis were performed at different time intervals confirming what the clinical course had already pointed out. These results show that SOD in its modified form, despite the lower dosage, can provide good protection against ischemia/reperfusion injury of muscles. PMID- 7499022 TI - Lentinan augments skin reaction induced by bradykinin: its correlation with vascular dilatation and hemorrhage responses and antitumor activities. AB - The effects of lentinan, an antitumor polysaccharide, on vascular reactions against vasoactive mediators were investigated in murine systems. Lentinan augmented intradermal reactions against bradykinin. Induction of acute phase proteins (APP) and the vascular dilatation hemorrhage (VDH) reaction on the ears have been reported to reflect the host responses to lentinan. The strain difference in the intensity of skin reactions coincided with those observed in VDH responses and with lentinan-induced antitumor effects against Sarcoma 180. Augmentation of skin reactions was not observed in T-cell-deficient mice. Inhibitors of lipoxygenase, thrombin and plasmin which reduced skin reactions also decreased the incidence of tumor necrosis positive mice among FBL-3-bearing mice treated with lentinan. Furthermore, B10D2 mice treated with fluorouracl (5 FU) and lentinan 10 days after S908.D2 transplantation showed complete tumor regression and augmented skin reactions, whereas augmentation of skin reactions and tumor regression were not observed in mice treated with 5-FU and lentinan 32 days after tumor inoculation. Taken together, these results suggest that these vascular reactions might play crucial roles in antitumor effects of lentinan and that the skin reaction, the convenient method for investigating vascular reactions, is a promising tool to monitor host sensitivity to lentinan in antitumor responses. PMID- 7499021 TI - Effect of exogenous leukotriene B4 (LTB4) on BALB/c mice splenocyte production of Th1 and Th2 lymphokines. AB - The effect of exogenous leukotriene B4 (LTB4) on the production of cytokines typical of Th1 (interleukin-2 and interferon-gamma) and Th2 (interleukin-4 and interleukin-10) lymphocytes was studied. Splenocytes were stimulated with concanavalin A (ConA) with or without different concentrations of LTB4 (3 x 10( 10) to 3 x 10(-7) M) for various times in the presence of BW 755C to inhibit the endogenous synthesis of eicosanoids. LTB4 was not able to induce cytokine secretion by itself. However, LTB4 augmented ConA spleen cell production of interleukin-2 (IL-2) and interferon-gamma (IFN-gamma) from Th1 cells and interleukin-4 (IL-4) and interleukin-10 (IL-10) from Th2 cells more than the controls treated with ConA alone. The pre-exposition of splenocytes to LTB4 for 3 h made these cells more sensitive to ConA in terms of IL-2 and IL-10 production than those treated with LTB4 at the onset of the incubation and maintained during the whole culture period. The results suggest that LTB4 may participate as a component of the signal transduction process for ConA-induced Th1 and Th2 cytokine production in a time-dependent manner. PMID- 7499019 TI - Anaesthesia and autologous transfusion. AB - The anaesthesiologist plays a central role in co-ordinating the combined application of the various blood saving techniques. In fact, to carry out transfusion therapy correctly, the anaesthesiologist must plan the right number of units of predeposit blood during the first examination, estimate the salvage of intra and post operative blood loss and spread the infusion of the units over the first three days in order to keep the patient in a state of haemodilution. From January 1992 to June 1994 in the department of anaesthesia and the intensive care unit, 980 patients were treated for total joint replacement: 714 total hips (7 after removal of plates and screws) 145 revisions, and 121 total knee prostheses. Basal Hb was 13.4 +/- 1.4 g/dl (range 6.7-17.9 g/dl). Homologous transfusions were carried out in 6.3% of these patients. The need to use homologous transfusions was negatively influenced by female sex, coronary heart disease (p = 0.005), length of surgery and type of antithromboembolic prophylaxis (indobufen has a significantly low incidence-p = 0.0001--compared to calcium heparin or low molecular weight heparin). PMID- 7499023 TI - Macrophage-like RAW 264 cell line and time-resolved fluoroimmunoassay (TRFIA) as tools in screening drug effects on cytokine secretion. AB - A screening system was set up to study the effects of drugs on cytokine secretion by macrophages in vitro. The system is based on the murine macrophage-like cell line RAW 264, which can be activated with lipopolysaccharide (LPS) to produce cytokines. The responsiveness of the RAW 264 cells was outlined by challenging them with different concentrations of LPS for 6 or 24 h. Substantial time- and dose-dependent increases were recorded for interleukin-1 beta (IL-1 beta), interleukin-6 (IL-6) and tumour necrosis factor alpha (TNF alpha) secretions. A general procedure was established to construct time-resolved fluoroimmunoassays (TRFIA) from commercial immunochemicals produced originally for enzyme immunoassays. Practical measuring ranges of the non-competitive assays were 100 pg/ml-10 ng/ml for IL-1 beta and TNF alpha and 10 pg/ml 5 for IL-6 and IL-5. The interleukin-5 (IL-5) assay was set up for unrelated human studies, but the others were used in the characterization of RAW 264 cytokine secretion. An immunosuppressive effect with dexamethasone phosphate could be achieved and recorded in the model system. Thus, the system offers a simple and easy-to-use model for screening immunomodulatory effects of drugs on the cytokine secretion of macrophages. PMID- 7499024 TI - Protective effect of leflunomide on the natural course of Leishmania major induced disease in genetically susceptible BALB/c mice. AB - Leflunomide has been reported as an immunomodulating agent which acts on a variety of cells including T- and B-lymphocytes. CD4+ T-lymphocytes are essential for the type of disease that develops after infection with the protozoan parasite Leishmania major. A variety of immunological interventions has been shown to modulate disease development. Therefore, the effect of leflunomide on the development of parasite-induced lesions and the ensuing immune response was investigated in genetically susceptible BALB/c mice. Oral feeding for 7 to 10 days of leflunomide (30 mg/kg per day) beginning 2 days prior to or at the day of infection led to the development of a stable resistant phenotype, i.e. to long lasting (> 13 months) regression of the lesions and clinical cure. Starting leflunomide treatment 3 days after infection was ineffective. The main bioactive metabolite, 1726 B, did not inhibit viability or growth of L. major promastigotes and amastigotes in vitro. Quantitative analysis of CD4+ and CD8+ cells in spleens and lymph nodes of parasite-infected animals treated with leflunomide for 5 days showed no significant effect. In vitro, 1726 B dose-dependently inhibited growth of stimulated T-cells, which could not be restored by saturating amounts of exogenous IL-2 and IL-4. No effect was observed on the killing function of activated macrophages. Taken together, the data indicate that leflunomide is a potent prophylactic agent to prevent an otherwise lethal infection of BALB/c mice. PMID- 7499025 TI - Experimental investigations on the immunomodulating activity of polybacterial preparation for peroral immunotherapy and immunoprophylaxis of uroinfections. AB - The immunomodulating effect of a polybacterial immunostimulator Urostim in experimental animal models was investigated. The preparation was created for immunotherapy and immunoprophylaxis of uroinfections. It consists of killed bacterial cells and their lysates of four microbial species: Escherichia coli expressing type 1 pili, Rc mutant of E. coli, Klebsiella pneumoniae, Proteus mirabilis and Enterococcus faecalis, BALB/c mice and guinea-pigs were treated orally with Urostim for 5 or 10 days, respectively. A stimulatory effect of Urostim on phagocytosis (increased phagocytic index and phagocytic number) of mice peritoneal exudate cells was observed. An increased number of plaque-forming cells and elevated titres of haemagglutinating antibodies in mice demonstrated activation of humoral immunity. Lymphoproliferative studies showed a significant response to phytohaemagglutinin (PHA) and Urostim, especially after the second application of the preparation. At the same time no changes in the absolute number of peripheral blood lymphocytes were found. An active protection of mice against a systemic infection and of guinea-pigs against uroinfection caused by Gram-negative bacteria was obtained. In conclusion, the results obtained characterize Urostim as an effective immunostimulator. Its peroral application leads to the activation of innate resistance, cell-mediated, humoral and local immunity. PMID- 7499026 TI - Inhibition of blood-brain barrier disruption in experimental allergic encephalomyelitis by short-term therapy with dexamethasone or cyclosporin A. AB - Double radioisotope measurement of neurovascular integrity in Lewis rats inoculated for experimental allergic encephalomyelitis (EAE) showed abnormal elevation of albumin extravasation in the cerebellum, medulla-pons and cervical spinal cord at the time of clinical manifestation. Therapeutically administered dexamethasone (Dex) (0.1-1 mg/kg body weight) or cyclosporin A (CsA) (25-75 mg/kg body weight) dose-dependently reduced albumin movement across the blood-brain barrier (BBB). Dex at a dose of 1 mg/kg completely suppressed abnormal BBB permeability in all tissues (P < or = 0.001), while CsA at the highest dose of 75 mg/kg achieved highly significant (P < or = 0.001), but not complete, suppression of aberrant barrier leakage in the areas studied. The implications of these findings to possible drug action at the immunocompromised cerebrovasculature are discussed. PMID- 7499027 TI - Hot water extracts of Chlorella vulgaris reduce opportunistic infection with Listeria monocytogenes in C57BL/6 mice infected with LP-BM5 murine leukemia viruses. AB - The bacterial elimination after infection with Listeria monocytogenes was impaired in mice with murine acquired immunodeficiency syndrome (MAIDS) by infection with LP-BM5 murine leukemia virus. Oral administration of hot water extracts of Chlorella vulgaris (CVE) restored the capacity of MAIDS mice to eliminate L. monocytogenes in association with improvement of the deteriorated immune response to L. monocytogenes. DTH response to Listeria in CVE-treated MAIDS mice was significantly higher than that of MAIDS mice after Listeria infection in association with increases in number of CD4+CD8- and CD4-CD8+ alpha beta T-cells in the infected sites. CVE might be effective in the treatment of opportunistic infection in retrovirus-induced immunodeficient patients. PMID- 7499028 TI - Adjuvant effect of polyadenylic.polyuridylic acid on antibody production of recombinant hepatitis B surface antigen in mice. AB - The adjuvant effect of polyadenylic.polyuridylic acid [poly(A).poly(U)] on antibody production against recombinant hepatitis B surface antigen (rHBsAg) was investigated. Immunization of adult Balb/c mice with rHBsAg plus poly(A).poly(U) elicited significantly higher antibody responses than immunization of mice immunized with rHBsAg alone. Such an adjuvant effect was evident even in mice sensitized with subimmunogenic doses of the antigen. PMID- 7499029 TI - Inhibition by cholesterylphosphorylserine of T-cell-mediated immune responses in mice. AB - The synthetic analogue of phosphatidylserine, cholesterylphosphorylserine (CPHS) inhibits T-cell-mediated immune responses in mice. Tested in cultured mouse spleen cells, CPHS inhibits concanavalin A-induced activation of DNA synthesis (IC50, 3.5 microM). Injected i.p. during the efferent phase, CPHS (25-100 mg/kg) inhibits the manifestations of delayed-type of hypersensitivity. The compound (25 mg/kg i.p., daily) reduces the acute graft-versus-host reaction when given for 5 days to donor mice before the isolation of spleen cells used for the inoculum. These data suggest that the addition of a phosphorylserine group to a steroid ring may produce immunoregulatory compounds. PMID- 7499030 TI - Mathematical modelling of HIV infection therapy. AB - The previously developed mathematical model simulates the CD4+ lymphocyte dynamics in HIV infection very well. As the number of these cells is a good indicator of the infection progression, it was used to evaluate the effectiveness of different therapeutic interventions. For chemotherapy simulation, both permanent and temporary zinovudine (AZT) administration were considered and the induced return of the CD4+ lymphocyte counts was analysed. Similar analysis was performed for active and passive immunotherapy. The model offers also the possibility of stimulating the CD4+ dynamics after depletion of CD8+ lymphocytes by antibodies. Even one simulated administration of anti-CD8 antibodies increases the CD4+ lymphocyte counts and prolongs the survival of the patient. However, if cells involved in protective immunity are assumed to belong to the CD8+ category, anti-CD8 antibodies accelerate the decrease of CD4- cells and thus shorten the patient's survival. PMID- 7499031 TI - Suppressive effects of volatile anesthetics on cytokine release in human peripheral blood mononuclear cells. AB - This study investigated the effects of three volatile anesthetics (sevoflurane, isoflurane, and enflurane) on cytokine release by human peripheral mononuclear cells (PBMCs) stimulated by natural killer (NK)-sensitive tumor cells, K562, in vitro. PBMCs, as effector cells, obtained from 31 volunteers were randomly allocated to two groups in the first set of experiments. One group was incubated with K562 (n = 21) and the other with medium alone as a control (n = 10). In a second set of experiments, PBMCs from each volunteer (n = 21) were divided into three groups: nonanesthetic, 1.5-MAC, and 2.5-MAC groups (n = 7 for each anesthetic). After 2 h exposure to anesthetic gas or air, K562 cells were added to the effector cells. After 4 h incubation, interleukin-1 beta (IL-1 beta), interleukin-2 (IL-2), tumor necrosis factor-alpha (TNF-alpha), and interferon alpha (INF-alpha) in the supernatant were assayed. IL-1 beta and TNF-alpha levels were significantly increased in comparison with those in the control group. IL-2 levels tended to be higher than those in the control group. No effect on IFN alpha levels was found. After anesthetic exposure, the releases of IL-1 beta and the release of TNF-alpha were significantly inhibited compared with those after air exposure. None of the anesthetics inhibited IL-2 release. The anesthetics studied are capable of altering the release of cytokines by NK and NK-like cells in response to tumor cells. PMID- 7499033 TI - Community rehabilitation workers: do they offer hope to disabled people in South Africa's rural areas? AB - This article outlines the thinking behind the Wits/Tintswalo community rehabilitation worker training programme which is located in South Africa's eastern Transvaal Lowveld. Drawing on a combination of qualitative and quantitative evidence collected during the first eighteen months in which the CRWs were working full-time (January 1993 - June 1994) it suggests that the two year training programme has been successful in providing therapeutic skills. Key issues raised are whether the coverage of disability is adequate in terms of numbers treated or in terms of categories of disability treated. PMID- 7499034 TI - Assessment methods for rehabilitation. AB - Diagnostics and evaluation in medical rehabilitation should be based on methods that are as objective as possible. In this context quantitative methods are an important precondition. We conducted for the German Pensions Insurance Institutions (which are in charge of the medical and vocational rehabilitation of workers and employees) a survey on assessment methods for rehabilitation which included an evaluation of American literature, with the aim to indicate procedures that can be considered for adaptation in Germany and to define further research requirements. The survey identified: (1) standardized procedures and instrumented tests for the assessment of musculoskeletal, cardiopulmonary and neurophysiological function; (2) personality, intelligence, achievement, neuropsychological and alcoholism screening tests for the assessment of mental or cognitive function; (3) rating scales and self-administered questionnaires for the assessment of Activities of Daily Living and Instrumental Activities of Daily Living (ADL/IADL Scales); (4) generic profiles and indexes as well as disease specific measures for the assessment of health-related quality of life and health status; and (5) rating scales for vocational assessment. German equivalents or German versions exist only for a part of the procedures identified. Translation and testing of Anglo-Saxon procedures should have priority over the development of new German methods. The following procedures will be taken into account: (a) instrumented tests for physical function, (b) IADL Scales, (c) generic indexes of health-related quality of life, (d) specific quality of life and health status measures for disorders of the circulatory system, metabolic system, digestive organs, respiratory tract and for cancer, and (e) vocational rating scales. PMID- 7499032 TI - Immunotoxicological screening of morphine and methadone in an extended 28 day study in rats. AB - Drug addicts are prone to infection with viruses including hepatitis-B and HIV. Besides indirect effects as a consequence of lifestyle, heroin and methadone may also enhance the risk of infections by a direct immunotoxic effect affecting resistance. In addition to general toxicological screening, we therefore performed a screening for potential immunotoxicity of morphine and methadone. Rats treated orally with different dosages of morphine or methadone for 6 weeks showed only a minor effect of overt toxicity on liver and spleen at the high dose, whereas at lower doses an increase in the relative weight of the mesenteric lymph nodes and an increase in cell density in the medullary cords were observed histopathologically, indicating a specific effect on humoral immunity. This specific immunotoxic effect was corroborated by an increased IgG concentration in serum (significant for the methadone-treated group). Further immunotoxicological research is needed aimed at revealing the potential risk of opiate use with respect to immune function. In conclusion, the present paper showed the toxicological profile of morphine and methadone in an extended 28 day subchronic study. Specific immunotoxicological effects were observed at doses where no effects were seen in routine toxicological evaluation, suggesting that the immune system is sensitive to opiates. PMID- 7499035 TI - Negative work behaviours: their impact on disabled people seeking work. AB - This paper looks at work behaviours which prevent an individual from getting and keeping a job, and then reviews how these behaviours might be changed. The term 'work behaviours' is examined and a definition for the term is proposed. Possible causes of negative work behaviours are cited, along with the way they might manifest themselves. A diagram demonstrating visually the causes and manifestations of negative work behaviours is shown. The potential for changing work behaviors is discussed, followed by a review of possible techniques that can affect such a change. Direct and indirect techniques of behaviour change are described. Direct behaviour change is seen to be preferable to indirect change, and the author looks briefly at nine methods of direct behaviour change. PMID- 7499036 TI - On the impediments to innovation in the European assistive technology industry. AB - This paper identifies and analyses some critical aspects of the European assistive technology industry which characterize innovation and determine its type, direction and diffusion. It is argued that innovation in this field is restricted because of (i) the limited and inefficient interaction between certain distinctive market elements, namely research, development, production, trade, service delivery, procurement and usage; (ii) the structural characteristics and general economic environment of the European assistive technology market; and (iii) the lack of a concise set of complementary and supportive actions towards the promotion of an environment favourable to industrial innovation and the transfer of new technology towards the European assistive technology market. The major conclusion from the investigation of the above factors is that innovation is not well served, while the limited innovative activity which is currently observed takes place mainly within institutions where the overall level of research seems to be concentrated and is of an occasional rather than permanent nature. Typically, this limited amount of innovation originates outside the assistive technology market and takes the form of 'induced' innovation. PMID- 7499038 TI - The self-employment option for people with disabilities: a case study of 'AHVA' desk top publishing company. AB - From the above results of the evaluation survey it can be concluded that the participants of the 'AHVA' desktop publishing company, who are with multiple and physical disabilities and who have initiated and run their own enterprise, perceived their first year of operation as beneficial. They mastered new technical skills, learned new work-related behaviours, expanded their interpersonal competencies and felt personal growth. They all developed high motivation to make their operation successful. The case study of 'AHVA' desktop publishing company, initiated and run by a group of multiply and physically disable persons, is unique and demonstrates the positive trend for the future. It is an example of personal and group commitment to build a competitive enterprise. It is self determination rather than the actual outcome of services provided, that is the driving force behind their efforts. The team members demonstrated that persons with disabilities can indeed take charge of their lives, be masters of their own destinies, and lead a productive and meaningful life. Here, inclusion is not a one-sided process in which service providers and rehabilitation experts try to integrate persons with disabilities into the community and regular employment. Inclusion is to integrate persons with disabilities into the community and regular employment. Inclusion is achieved here by the self initiative of a group of persons with disabilities. They have created an extraordinary enterprise according to their own special needs. Non disabled people interact with them on an equal level, as partners in a business, sharing mutual interests of quality production.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7499039 TI - Can activity monitors be used to assess compliance with walking programmes in the elderly? PMID- 7499037 TI - Characteristics of patients from hospital-based in-patient rehabilitation in The Netherlands. AB - The University Hospital Dijkzigt in Rotterdam is one of the first hospitals in The Netherlands to have a rehabilitation department providing multidisciplinary, hospital-based-in-patient rehabilitation (HBIR). The main goals of HBIR are to estimate the final optimum level of patient's functioning, to investigate the most appropriate treatment setting to achieve this and to fulfil requirements for discharge from hospital as soon as possible. One of the central issues in rehabilitation in the Netherlands is to obtain more insight into the current daily practice of HBIR. The aim of this study was to describe the characteristics of the patients treated in HBIR in Dijkzigt Hospital. Registered data from 1967 HBIR patients treated between 1988 and 1990 were analysed retrospectively. The patients were on average 53.1 ( +/- 19.7) years old; the men (63%) were significantly younger than the women (mean age of 50.8 and 57.0 years, respectively). The main diagnostic groups were stroke (27%), progressive and regressive neurological conditions (17%), hand injuries (9%), spinal cord lesion (9%), orthopaedic injuries (8%) and amputation of lower extremity (8%). The median duration of HBIR and hospitalization were 13 days and 33 day, respectively. Most patients were discharged home (63%), or to a nursing home (14%) or a rehabilitation centre (8%). The results of this study can be used in policy discussions on HBIR and provides a description of the population which needs to be studied longitudinally in future research on the outcomes of HBIR. PMID- 7499041 TI - Factor structure of the REHAB scale in an Irish sample. PMID- 7499040 TI - Rehabilitation after total hip replacement. PMID- 7499042 TI - Shoulder pain after a stroke. PMID- 7499043 TI - Poorly differentiated pediatric head and neck neuroblastoma: a diagnostic dilemma. AB - Primary pediatric head and neck neuroblastoma is rare, with metastatic disease being the more common mode of involvement in this anatomic region. Poorly differentiated neuroblastoma initially presenting in head and neck locations presents a diagnostic dilemma, especially when evidence of primary disease in typical abdominal, retroperitoneal and thoracic sites is lacking. This tumor cannot easily be distinguished from the other common pediatric small round cell malignancies that may originate in or metastasize to the head and neck. Recent years have seen great strides in the immunohistochemical as well as cytogenetic characterization of certain pediatric small round cell lesions. A never before reported solitary, poorly differentiated neuroblastoma of the right parotid gland in a 20-month-old female is presented in order to familiarize the otolaryngologist with the modern diagnostic armamentarium available for the accurate characterization, and thus appropriate workup and treatment of this disease. PMID- 7499044 TI - Pediatric tracheotomy discharge teaching: a comprehensive checklist format. AB - Discharge planning for a child undergoing a tracheotomy is a complex process. In 1989, a multidisciplinary team at Children's Hospital of Michigan developed specific discharge criteria in a checklist format to address all facets of home care for these patients. We present and discuss the checklist. A survey of user satisfaction with the checklist demonstrated that 80% of parents and care-givers felt well-prepared by this format at the time their child was discharged from the hospital with a new tracheotomy. We conclude that the protocol allows for comprehensive and efficient discharge teaching of parents and care-givers for children with new tracheotomies. PMID- 7499045 TI - Treatment of experimental acute otitis media with ibuprofen and ampicillin. AB - The efficacy of concurrent treatment of experimental acute otitis media with ibuprofen and ampicillin was evaluated in chinchillas with respect to clearance of the effusion and resolution of mucosal inflammation. Sixty-four chinchillas were infected with Streptococcus pneumoniae and randomly assigned to treatment with either IM ampicillin (control) or ampicillin plus ibuprofen (experimental) beginning on day 2 post inoculation. On days 3, 6, 9 and 12, 8 animals from each group were killed, effusions recovered for biochemical assay and the right middle ears prepared for histological study. Between group differences in the number of ears with effusion and effusion volume were not statistically significant. Mucosal thickness and the frequencies of ears with histopathological signs of inflammation were significantly less in the experimental group when compared to the control group. Differences in the effusion concentrations of total protease, 3 of 4 hydrolytic enzymes and free fatty acids favoring the experimental group were observed at the 6, 9 and 12 day endpoints. Also, at those times the levels of the 3 measured products of the cyclooxygenase pathway were less in the experimental group. These results suggest that the addition of ibuprofen to ampicillin for the treatment of acute otitis media decreases production of select eicosonoids, reduces mucosal inflammation and alters the course of the disease in this model of bacterial infection. PMID- 7499046 TI - Tonsillectomy and adenoidectomy in patients with Down syndrome. AB - Tonsillectomy and adenoidectomy in patients with Down syndrome usually relieves symptoms of upper airway obstruction due to adenotonsillar hypertrophy. A retrospective review of 16 patients with Down syndrome who underwent a tonsillectomy and adenoidectomy was conducted to define early postoperative morbidity, and to evaluate the results of the procedure. Fifteen of 16 patients had upper airway obstruction manifest by adenotonsillar hypertrophy. One had adenoid hypertrophy only. Tonsillectomy and adenoidectomy (13), adenoidectomy (1), tonsillectomy (1), and UP3/adenoidectomy (1) were performed on these patients. Four patients (25%) required observation in an intensive care setting the first night. An average hospital stay of 2.1 days was noted with a range of 0 to 7 days. Significant postoperative apnea was common, and oxygen was used in over 60% of patients. Symptoms were resolved in 69% of patients at last follow up. In conclusion, tonsillectomy and adenoidectomy can be a useful procedure for children with Down syndrome and obstructive sleep apnea, however overnight hospitalization for observation and treatment of persistent apnea is appropriate. PMID- 7499047 TI - Bifid epiglottis syndrome. AB - True bifid epiglottis is an extremely rare laryngeal anomaly, which usually presents in the neonate with symptoms of aspiration and/or airway obstruction. Management is generally supportive observation as the symptoms lessen with age, but rarely tracheotomy is required for airway obstruction. Bifid epiglottis occurs in a syndromic picture with associated anomalies, especially polydactyly, cleft palate and retro/micrognathia but a significant number will have endocrine, gastrointestinal and genitourinary abnormalities. A case of true bifid epiglottis is presented and the literature is reviewed on the subject. PMID- 7499048 TI - Unusual complication of temporomandibular joint surgery: cervicomediastinal abscess. AB - Cervicomediastinal abscess developed in a 14-year-old girl after temporomandibular joint surgery. No such complication could be found in the literature. The complications of temporomandibular joint surgery and the possible ways this abscess may have developed are discussed. PMID- 7499049 TI - Pleomorphic adenoma of an unusual size in the deep lobe of the parotid gland. AB - Tumors of the salivary glands in children are not as common as in adults and they are mostly benign. The most common benign epithelial neoplasm seen in children is pleomorphic adenoma. As is usually the case in the adult population, it is usually found in the superficial lobe of the parotid gland. Its localization in the deep lobe is rare. Pleomorphic adenomas of the deep lobe present with a neck mass and when it is large, it may have an intraoral extension as well. It may also assume a dumb-bell shape as it enlarges and may be observed on CT or MRI scans as a prestyloid mass. The choice of treatment depends on the localization and the size of the tumor. The method can be transoral, cervical, transmandibular, transparotid, retromandibular fossa or a combination of these. A child with an unusually large pleomorphic adenoma of the deep lobe of the parotid gland is presented in this study. The deep lobe tumor was totally removed using a combination of cervical and transmandibular approaches. PMID- 7499051 TI - Juvenile hyaline fibromatosis. AB - A case of juvenile hyaline fibromatosis in a 7-year-old Asian boy is presented. This autosomal recessive inherited condition has not been described in the otolaryngology literature before. We demonstrate the benefits surgical intervention, for treatment of gingival hypertrophy, can bring to the patient and outline the other features of this rare condition. The literature on juvenile hyaline fibromatosis and infantile systemic hyalinosis is also reviewed. PMID- 7499050 TI - Fibrous hamartoma of the tongue: report of a case with immunohistochemical and ultrastructural studies. AB - The case of an 8-month-old female with fibrous hamartoma of the tongue is reported with immunohistochemical and electron microscopic findings. An elastic hard mass at the base of the tongue gradually enlarged, causing dysphagia. The tumor was excised by CO2 laser. Under light microscopy, the tumor was observed as composed of fibroblast-like cells, fibrocollagenous bundles and striated muscle tissues. The majority of the cells were immunoreactive for vimentin. Electronmicroscopically, they contained numerous intermediate filaments, some had intercellular microvilli and cilia (9 + 2 type). These findings revealed that the cells of the fibrous hamartoma in our case may possess both mesenchymal and epithelial properties. PMID- 7499052 TI - Language--the outcome measure for the linguistically developing cochlear implant patient. PMID- 7499053 TI - Mental health nurses working purposefully with people with serious and enduring mental illness--an international perspective. AB - There has been a world-wide refocus on people with serious and enduring mental health problems. This has provided major challenges for all those involved in mental health care, as the last two decades have produced radical changes in service delivery and a range of new interventions. Mental health nurses are playing a central role as community based clinical case managers and are beginning to use a range of psychosocial interventions. They are also revitalizing skills in the more traditional areas of medication management and residential care. In addition, there are growing problems of substance abuse and violence which needs to be addressed. Mental health nursing is responding in this new era by various training initiatives. However, these programmes need rapid expansion as mental health nurses will continue to play a key role in the services of the future and therefore need these new skills. PMID- 7499054 TI - The current status and future challenges of psychiatric/mental health nursing. AB - To obtain a global picture of the current status of nursing education and nursing practice in the mental health/psychiatric field, the International Council of Nurses (ICN) sent a questionnaire to 23 of its 101 member associations. The basis for the selection was to obtain information from a broad spectrum of different associations, e.g. state of development--economic and political, language, and geographical region. Of these, 16 (or 69%) returned completed questionnaires, often accompanied by extensive supporting documentation. In addition, four nurses' associations in Africa completed the questionnaire during staff field visits in that region. This paper makes reference to the responses received to the questionnaire and some verbatim quotes are included. PMID- 7499055 TI - Recent developments in psychiatric nurse education in the countries of central and eastern Europe. AB - Since the social and political revolutions of 1989 and the years that followed the countries of Central and Eastern Europe (CCEE) have undergone radical changes in their health care systems, and among the most radical of these has been the restructuring and reorganisation of the psychiatric nurse training and education, with a weather eye to the potential of mental health demands that may result from the socio-economic turmoil. This paper, much of which was compiled while on secondment to the WHO, attempts to give a brief overview of the main policy developments in selected countries and to place them within a philosophical, social and pedagogical context. It cannot escape notice that the way in which countries choose to educate their nurses and what they can be expected to achieve in their work, reflects a more global understanding of mental health--and that may be why we should all learn from this experience. PMID- 7499057 TI - The proper focus of nursing: a critique of the "caring" ideology. AB - Some nurse educators in the United Kingdom are following the lead of North American theorists by asserting that caring "is" the core of nursing. This paper offers a focused critique of the caring ideology, through an examination of: (a) the views of a founding figure in the "caring" movement, and (b) a proposal for the integration of a "caring curriculum" within Project 2000. An overview of the "caring" literature prefaces an examination of Watson's notion of a "science of caring". Finally, Kirby and Slevin's recent proposals for a "curriculum of care" are examined in terms of their necessity and practicality. PMID- 7499056 TI - Working in partnership with people who use services; reaffirming the foundations of practice for Mental Health Nursing. AB - This paper reports on work produced by a Government Review of Mental Health Nursing in England conducted in 1993/4. The Review reported to the British Government in January 1994 and offered 42 recommendations for their consideration. At the heart of the Review Report was a determination that patients, carers and all people who use services should be engaged in a new partnership with nurses in the delivery of care. Within the 42 recommendations there are a number which relate particularly to developing this partnership. This paper discusses these recommendations and the issues which surround them. PMID- 7499058 TI - The tasks of psychiatric nurses in long-term residential settings in The Netherlands. AB - The tasks of the psychiatric nurse, working in long-term psychiatric settings, are described and analysed in this study. A questionnaire was completed by 361 nurses in five different psychiatric hospitals in the Netherlands. Nurses appear to concentrate upon personal care tasks in and around their ward. This does not fit adequately the changes in psychiatric practice. By means of an externally oriented hospital policy, cooperation with other care agencies and a realization of a more patient-orientated and rehabilitation model of care, the nurse is better prepared for helping patients make their move to the community. PMID- 7499060 TI - Effect of pyrrolidone carboxylate (PCA) and pyridoxine on liver metabolism during chronic ethanol intake in rats. AB - Rats subjected to chronic ethanol intake for a period of 28 days showed significant elevation in blood ethanol levels, a marked decrease in hepatic reduced glutathione (GSH) content and a decrease in liver tryptophan pyrrolase (TPO) activity. A daily intraperitoneal injection of a combined solution of pyrrolidone carboxylate (PCA) and vitamin B6 (pyridoxine hydrochloride) (0.3 mmoles/kg) into ethanol-treated rats resulted in the blood ethanol levels becoming significantly reduced, while the hepatic GSH content and TPO activity were markedly elevated. Our results support the view that PCA and pyridoxine operate to restore the redox imbalance of the hepatocytes caused by chronic alcohol intake. PMID- 7499059 TI - Antiplatelet activity of synthetic and natural resveratrol in red wine. AB - The antiaggregating effect of the phytoalexin resveratrol (3,4,5 trihydroxystilbene), alone or associated with red wine, and polyphenol have been evaluated in vitro at different concentrations on platelet-rich plasma from healthy volunteers. Resveratrol at the concentration of 3.56 micrograms/l was able to lower platelet aggregation by 50.3% +/- 1.83. Red wine containing 1.2 mg/l of natural trans-resveratrol and 3.6 milligrams of polyphenols diluted 1000 fold (final resveratrol concentration: 1.2 micrograms/l) inhibited platelet aggregation by 41.9% +/- 2.11. By adding resveratrol to the wine up to a concentration of 1.2 micrograms/l, inhibition was raised to 78.5% +/- 4.70. These results suggest that the antiaggregating activity of resveratrol is related to its concentration in wine. PMID- 7499061 TI - Effect of L-propionyl carnitine on some properties of erythrocytes and leukocytes of alcohol abusers. AB - The effect of L-propionyl carnitine, the carnitine derivative utilized as a more effective drug for membrane protection, on Na-K ATPase activity of erythrocyte ghosts of alcohol-dependent patients and blood donors has been investigated. The effect of L-propionyl carnitine on leukocyte chemotaxis and cytochrome c reduction, a measure of superoxide ion production was also studied. It has been in fact observed that alcohol is immunotoxic on both the non-specific and the specific immune response. In alcohol-dependent erythrocytes, a significant higher value of the 1H-NMR spin lattice relaxation time (T1) was observed as compared to blood-donor erythrocytes. The in-vitro addition of ethanol increases the T1 values of blood donor erythrocytes, whereas it is without effect on the T1 value of alcohol-dependent erythrocytes. The Na-K ATPase activity is higher in alcohol dependent erythrocyte ghosts as compared to blood-donor ghosts. A non-significant increase of the Na-K ATPase activity of blood-donor ghosts was observed with the increasing of L-propionyl carnitine concentrations (from 0.2 to 5.0 mM), whereas the Na-K ATPase activity of alcohol-dependent ghosts decreases. From these combined effects the differences of Na-K ATPase activity progressively decrease with the increasing of L-propionyl carnitine concentration, and no significant differences are observed between the two groups at L-propionyl carnitine concentrations higher than 0.5 mM. The in-vitro addition of ethanol increases the enzyme activity to a greater extent in blood-donor ghosts as compared to alcohol dependent ghosts. This in-vitro activation by ethanol is decreased by the addition of L-propionyl carnitine. The chemotaxis induced by N-formyl-methionyl leucylphenylalanine and the superoxide anion production stimulated by zymosan is significantly lower in alcohol-dependent neutrophils. L-Propionyl carnitine increases, in a dose-dependent way, both chemotaxis and superoxide anion production of alcohol-dependent neutrophils, and no significant difference was observed between the two groups at 5 mM L-propionyl carnitine. These experimental results suggest that L-propionyl carnitine administration may be useful for reducing some acute and chronic damages due to alcohol ingestion. The protective and modulatory actions of L-propionyl carnitine may be even more evident in cells and tissues different from those investigated in this study and in which ethanol determines several biochemical damages. PMID- 7499062 TI - Joint failure in erosive osteoarthritis of the hands. AB - Erosive osteoarthritis (EOA), the type of hand osteoarthritis characterized by faster destructive changes, may be suspected clinically because of phlogistic presentation especially in DIP and in PIP joints. Scintiscan may be useful in early diagnosis, but the final diagnosis of EOA is always radiological, and depends upon the presence of central erosions. The inflammatory and crystal induced origin of EOA is sometimes stressed in the literature, but despite the phlogistic course of the disease there is evidence against EOA being a primitive synovitis, the most cogent point being the cellular content of synovial fluid. The pathogenetic hypothesis that we propose is of a chondral primitive disorder, with synovitis causing further chondral erosion, subchondral bone damage, and finally joint instability or fibrous and bone ankylosis with severe impairment of joint function. PMID- 7499063 TI - Acute effects of calcitonin gene related peptide on renal haemodynamics and renin and angiotensin II secretion in patients with renal disease. AB - The renal haemodynamic effects and renin-angiotensin II response to calcitonin gene-related peptide (CGRP) infusion were assessed in 16 patients with moderate hypertension and renal insufficiency. CGRP lowered the systemic mean blood pressure by 13% and increased the heart rate by 25%; the glomerular filtration rate rose from 56 +/- 11 ml/min to 71 +/- 8 ml/min (p < 0.005), the renal plasma flow decreased from 369 +/- 19 ml/min to 342 +/- 25 ml/min (p < 0.002) and the filtration fraction increased from 15 +/- 0.2% to 20 +/- 0.2%. Plasma renin activity rose stepwise during the CGRP infusion from 1.28 +/- 0.5 ng/ml/h to 1.66 +/- 0.4 and 1.89 +/- 0.4 ng/ml/h (p < 0.001). Angiotensin II showed a marked increase after 10 min of infusion (91.6 +/- 47.00 pg/ml) (control value 6.01 +/- 3.09 pg/ml) and at the end (28.63 +/- 16.00 pg/ml) (p < 0.001). CGRP exerts an apparently favourable effect on renal function of patients with renal insufficiency, but the observed increase of glomerular filtration rate is obtained by an increase of intraglomerular pressure secondary to angiotensin II production. PMID- 7499064 TI - Alteration of spleen lymphocyte populations in rats with arthritis induced by muramyl dipeptide analogue or complete adjuvant. AB - To examine the involvement of lymphocytes in the development of MDP-Lys(L18) induced arthritis (MIA) in rats and the exacerbation of MIA by cyclosporin A (CsA), we analysed the spleen lymphocyte subset using monoclonal antibodies and flow cytometry during the development of arthritis and compared the results with those found in adjuvant-induced arthritis (AIA). Subcutaneous injection of MDP Lys(L18) 4 mg/kg to male Lewis rats for 14 days caused very slight and quite clear increases in tarsal joint thickness on days 8 and 15, respectively. This increase was significantly enhanced by co-administration of CsA 10 mg/kg on both of these days. Adjuvant intracutaneously injected once increased the thickness only on day 15, and this was completely inhibited by CsA. The populations of CD4+ and CD8+ cells were increased and decreased, respectively, increasing the CD4+/CD8+ ratio, from day 8 in MIA. CsA enhanced the MDP-Lys(L18)-induced changes in these populations and caused additional decreases in the number of CD5+ cells. Only the CD4+ cell population was increased on day 15 in AIA, and this increase was inhibited by CsA. These results suggest that the spleen lymphocyte subsets in MIA have a different role from those in AIA, and that the contribution of enhancement of the subset changes to the exacerbating effect of CsA on MIA. PMID- 7499065 TI - [Asthma therapy with topically active steroids]. PMID- 7499066 TI - [What are the recommendations for prevention in patients at high risk for hyperthyroidism when an examination with iodinated contrast media becomes necessary?]. PMID- 7499067 TI - [What is collagen colitis and how is it treated?]. PMID- 7499068 TI - [Comment on the interview with R. F. Schmidt and G. Thews. Clinical principles and medical decision making]. PMID- 7499069 TI - [Cyanosis]. PMID- 7499070 TI - [Epidemiology of tuberculosis]. PMID- 7499071 TI - [Diagnosis of tuberculosis]. PMID- 7499072 TI - [Clinical aspects and therapy of tuberculosis]. PMID- 7499073 TI - [HIV infection and tuberculosis]. PMID- 7499074 TI - [Epidemiology of multidrug-resistant tuberculosis in the United States of America]. PMID- 7499075 TI - [Clinical aspects and therapy of ubiquitous mycobacterioses]. PMID- 7499076 TI - [24-year-old patient with acute abdomen and ascites]. PMID- 7499077 TI - [25-year-old patient with pancytopenia]. PMID- 7499079 TI - Keratocyte gap junctional communication in normal and wounded rabbit corneas and human corneas. AB - PURPOSE: Several studies have indicated the anatomic and biochemical presence of gap junctions in corneal keratocytes. The current study was designed to demonstrate that these gap junctions are functional in rabbit and human corneal keratocytes. This study also examined dye coupling between keratocytes migrating into the wound region of freeze-wounded rabbit corneas. METHODS: Freeze wounds were created on anesthetized rabbit corneas using a liquid nitrogen-cooled brass probe. Freeze-wounded corneas were examined at several time periods from days 0 to 5 after wounding. Nonwounded rabbit corneas also were examined. Human corneal buttons were examined immediately after removal from patients who underwent keratoplasty. Gap junctional coupling was examined by microinjecting carboxyfluorescein from microelectrodes into the basal-most keratocytes and capturing dye spread images with a cooled charge coupled device camera. RESULTS: Significant dye spread was observed between cells in the unwounded areas of corneas at wound time 0 and between cells migrating into the wound areas as early as 24 hours after wounding. In control corneas, dye spread to as many as 50 cells from the source cell. Dye spread also was seen between keratocytes in human corneas with pseudophakic bullous keratopathy and keratoconus. CONCLUSIONS: Gap junctions observed in keratocytes from normal rabbit corneas are functional. Gap junctions also are present and functional in keratocytes within unwounded and wounded regions of freeze-injured corneas. In addition, functional gap junctions are present between keratocytes in human corneas. This study confirms the long held contention that corneal keratocytes form a large intercommunicating network within the corneal stroma. PMID- 7499078 TI - Novel aspects of the ultrastructural organization of human corneal keratocytes. AB - PURPOSE: Proper functioning of the endothelium and proper structural organization of the keratocytes and collagen bundles are of ultimate importance for transparency of the cornea. The role of the endothelium has been investigated extensively, whereas the role of the keratocytes is still unclear. Detailed knowledge on the ultrastructural organization of keratocytes and the relationship between keratocytes and collagen bundles is as essential for understanding corneal transparency as is knowledge of endothelial functioning. METHODS: Thirty five corneas (30 postmortem donor corneas and 5 fresh corneas from the operating theater; age range, 28 to 90 years) were used for light microscopy, transmission electron microscopy, and scanning electron microscopy. Serial frontal sections of the central stroma reaching from epithelium to endothelium and cross-sections were studied. At three levels, reconstructions of the mutual arrangement of keratocytes were made using semithin sections. RESULTS: Keratocytes have the appearance of highly active cells with an abundancy of organelles. Between the dendritic ramifications of these cells, large amounts of amorphous material is observed. One of the most remarkable observations is the presence of an extensive network of fenestrations along the surface of the keratocytes. Another important observation is the circular arrangement of keratocytes gradually turning clockwise like a corkscrew from epithelium to endothelium. CONCLUSIONS: From the current study, the following conclusions can be drawn: Keratocytes are not quiescent but are highly active cells probably involved in turnover of the extracellular matrix; fenestrations may be of functional relevance with respect to facilitation of diffusion and mechanical attachment of the collagen fibers to the keratocytes; the corkscrew organization of keratocytes suggests that they form completely closed sheets of communicating cells throughout the depth of the cornea, creating equal chances for all light rays to pass one or more keratocytes and thus minimizing variation in light scattering over the entire cornea. PMID- 7499081 TI - Development of cytoskeleton in neuroectodermally derived epithelial and muscle cells of the human eye. AB - PURPOSE: To analyze the cytoskeletal development in neuroectodermally derived epithelial and muscle cells of the human eye during the second and third trimesters of pregnancy. METHODS: Nine formalin-fixed, paraffin-embedded fetal autopsy eyes from the 13th to 40th week of gestation were studied with eight monoclonal antibodies (mAbs) to intermediate filaments and alpha-smooth muscle actin (alpha SMA) by the avidin-biotinylated-peroxidase complex method. RESULTS: The epithelium of the iris reacted with mAbs Vim 3B4 and V9 to vimentin in all eyes. Reactivity with mAb CAM 5.2 to cytokeratin (CK) 8 and mAb CY-90 and KS B17.2 to CK 18 disappeared from the iris epithelium by the 28th gestational week. mAb 1A4 to alpha SMA labeled its anterior layer from the 28th week on. mAbs DE-U 10 and D33 to desmin labeled dilator fibers by the 37th week, and focal reactivity for CK 8 and 18 concurrently appeared. The developing iris sphincter reacted for alpha SMA in all eyes. It was labeled increasingly for desmin from the 18th week on, whereas initial reactivity for vimentin gradually disappeared after the 22nd week. mAbs to vimentin, CK 8, and CK 18 labeled the ciliary epithelium and the retinal pigment epithelium in all eyes, except for lack of CK 8 and 18 in the nonpigmented ciliary epithelium at the 13th week. The ciliary muscle reacted uniformly for vimentin and alpha SMA and from the 16th week on for desmin. CONCLUSIONS: The results highlight the individuality of cytoskeletal profiles in the neuroectodermal epithelial and muscle cells of the eye and clarify the formation of the peculiar cytoskeleton typical of the adult human eye. They also offer a framework for detecting cytoskeletal changes in developmental anomalies of the eye after the first trimester of pregnancy. PMID- 7499080 TI - Impact of vitamin A supplementation on prevalence and incidence of xerophthalmia in Nepal. AB - PURPOSE: To assess the impact of vitamin A supplementation at 4-month intervals on the prevalence and incidence of xerophthalmia among preschool-age children. METHODS: A stratified, random sample of 40 wards with 4766 children in Sarlahi district of Nepal was selected to participate in a randomized, controlled, community trial. In the vitamin A group, at 4-month intervals, neonates received 50,000 IU, 1- to 11-month-old infants received 100,000 IU, and children 1 through 4 years of age received 200,000 IU. Children underwent eye examination before the intervention and 16 months later. RESULTS: Before the intervention, 4318 children were examined for xerophthalmia. The prevalence was 2.3% in the vitamin A group and 3.3% in the placebo group. All children with xerophthalmia were treated with vitamin A at the time of the examination. Of those examined at baseline, 38 in the vitamin A group and 48 in the placebo group died in the 16 months after intervention. There were 1871 (84%) surviving children in the vitamin A group and 1711 (85%) in the placebo group examined at follow-up. After adjustment for the baseline prevalence of xerophthalmia, vitamin A reduced the prevalence at follow up by 63% (95% confidence interval, 21% to 83%). The apparent incidence was 3.2/1000 per year in the vitamin A group and 9.2/1000 per year in the placebo group, an adjusted reduction of 62% (95% confidence interval, 0% to 86%). CONCLUSIONS: Supplementation was effective at reducing the prevalence and incidence of xerophthalmia. PMID- 7499082 TI - Pathology of the iridocorneal-endothelial syndrome. The ICE-cell. AB - PURPOSE: The iridocorneal-endothelial (ICE) syndrome is characterized by glaucoma, corneal failure, and iris destruction. Specular photomicroscopy of the corneal endothelium in this disease shows a population of abnormal cells named ICE-cells. Comparison between ultrastructural examination and specular photomicroscopy demonstrates that the histologic equivalent of ICE-cells are cells with an epithelial phenotype. The authors have studied the differentiation markers expressed by ICE-cells using an ultrastructural immunocytochemical technique. METHODS: Seven keratoplasty specimens from patients with the ICE syndrome were examined by scanning and transmission electron microscopy and light and electron microscopic immunocytochemistry. Comparison was made with three normal corneas. Immunocytochemical studies were performed with monoclonal antibodies to broad-spectrum cytokeratins, cytokeratins 3, 5/8, 8/18 and 19, vimentin, and epithelial membrane antigen. RESULTS: ICE-cells were morphologically similar to epithelial cells and expressed the same profile of differentiation markers as did normal limbal epithelial cells. CONCLUSIONS: ICE cells may arise from an embryologic ectopia of ocular surface epithelium. Alternatively, these findings are consistent with a metaplastic stimulus resulting in a profound change in the phenotype of normal corneal endothelial cells. PMID- 7499083 TI - Function and cell surface phenotype of dendritic cells from rat cornea. AB - PURPOSE: To isolate dendritic cells (DC) from rat corneas and to examine their functions and surface markers in vitro. METHODS: Cells were isolated enzymatically from dissected rat corneas and cultured for various intervals of time. Dendritic cells were enriched immunomagnetically from corneal cell preparations using monoclonal antibodies against DC surface antigens and tested for functional activity in lymphocyte stimulation assays. In vivo migration of DC was induced by traumatizing the corneal epithelium. Wholemounts of epithelial sheets were stained immunofluorescently with anti-DC antibodies and examined by confocal microscopy. Dendritic cells isolated from traumatized corneas were tested for functional activity. RESULTS: Corneal DC exhibited the properties of other members of the DC family, i.e., low buoyant density, lymphoid DC-specific markers, and lymphostimulatory function. In fresh unfractionated cell preparations of normal cornea, no functional activity was detected. However, DC immunomagnetically purified from fresh preparations were functionally active. Injury to the corneal epithelium induced the migration of DC from the periphery to the central cornea; DC measured in this situation showed significantly increased functional activity. Finally, IL-1 beta and GM-CSF enhanced the functional activity of corneal DC. CONCLUSIONS: Corneal DC have lymphostimulatory capacity in situ, but they may be maintained in a state of latency by the suppressive influence exerted by neighboring cells. Injury to the corneal epithelium results in functional activation of the corneal DC, which may be caused by cytokines such as IL-1 beta or GM-CSF. Thus, corneal DC may be important in the immune regulation of the anterior segment. PMID- 7499084 TI - ETB and epidermal growth factor receptor stimulation of wound closure in bovine corneal epithelial cells. AB - PURPOSE: To determine if there is a heterogeneous pattern of endothelin (ET) receptor subtype (i.e., ETA and ETB) gene expression in the bovine corneal epithelium (BCE). To determine if ET receptor subtype stimulation increases the effectiveness of epidermal growth factor (EGF) to accelerate wound closure in a primary culture of bovine corneal epithelial cells (BCEC). METHODS: In situ hybridization histochemistry was used to characterize ETA and ETB gene expression in the BCE. A wound closure assay evaluated wound healing rates in BCEC after 4 to 7 days in culture. [3H] thymidine incorporation and MTT assay measured proliferation. RESULTS: ETA gene expression was appreciably higher in the basal cells than in the suprabasal cells, whereas the pattern for ETB was reversed. Epidermal growth factor (5 ng/ml) maximally increased wound closure by 145% above the control. With 5 ng/ml EGF, either 10(-9) M ET-1 or 10(-8) M sarafotoxin-6-c (s-6-c) increased wound closure by an additional 39% (P < 0.001) above that measured with 5 ng/ml EGF alone. BQ123 (10(-7) M) did not alter any of these effects of ET-1 or s-6-c. Epidermal growth factor stimulated wound closure through a selective increase in proliferation. Neither ET-1 nor s-6-c alone had any effect on proliferation or migration. CONCLUSIONS: Both ETA and ETB genes are expressed in BCE. However, in BCEC only, ETB stimulation increases the effectiveness of EGF to stimulate wound closure. This response was caused by an increase in cell migration rather than proliferation because, after treatment with mitomycin C, neither ET-1 nor EGF stimulated wound closure. PMID- 7499085 TI - Ocular resurfacing and alloepithelial rejection in a murine keratoepithelioplasty model. AB - PURPOSE: To determine definitively the epithelial origin of corneal resurfacing and to elucidate the immunologic mechanisms of epithelial rejection in a murine keratoepithelioplasty (KEP) model. METHODS: After corneal epithelial removal and peritomy, donor corneal lenticules were grafted around the limbus (KEP procedure). The process of corneal reepithelialization was observed with 0.25% methylene blue staining. The origin of the renewed epithelium was determined by immunofluorescence. Syngeneic corneal lenticules were grafted to BALB/c mice. C3H/He, C57BL/6, BALB.K, DBA/2, and B10.D2 allogeneic corneal lenticules were grafted to BALB/c mice, and A.SW and A.TL allogeneic corneal lenticules were grafted to A.TH mice. Alloepithelial rejection was evaluated on the basis of clinical findings and histologic changes in grafted corneas. RESULTS: All KEP grafts were reepithelialized entirely at 3 days after surgery. The renewed epithelium proved to be derived from the lenticules in BALB/c eyes receiving C3H/He lenticules. In syngeneic grafts 5 days after KEP, the cornea recovered clarity and smoothness, which persisted to the end of the study. After complete reepithelialization, all allogeneic grafts also experienced a short duration of clear cornea. Then followed four characteristic phases of inflammatory epithelial response: initial phase, acute phase, chronic phase, and rejected phase. Histologic examination confirmed the progress and severity of inflammatory response. The mean onset times of initial phase in assorted grafts with mismatched histocompatibility antigens were: 7.9 +/- 1.8 days for both major and minor disparity grafts, 9.5 +/- 3.8 days for major disparity grafts, 18.2 +/- 5.5 days for major histocompatibility class I disparity grafts, 25.6 +/- 7.2 days for major histocompatibility class II disparity grafts, and 9.2 +/- 2.2 days for multiple minor disparity grafts. CONCLUSIONS: In donor corneal lenticule grafting to host eyes with corneal epithelium removed and conjunctival peritomy, the ocular surface was reepithelialized by lenticule-derived epithelium. Alloepithelial rejection in this model displayed characteristic manifestations and well-defined processes, enabling easy and precise evaluation of onset and intensity of graft rejection. Both major and minor histocompatibility antigens are related to corneal epithelial rejection. PMID- 7499086 TI - The appearance of rod opsin during monkey retinal development. AB - PURPOSE: To determine the temporal and spatial pattern of rod opsin appearance in Macaca monkey retina. METHODS: Frozen sections from fetal day (Fd) 55 to adulthood (birth = Fd168) containing the entire horizontal meridian were stained using Rho4D2 monoclonal antiserum visualized with immunofluorescent labeling. At Fd66, Fd79, and Fd89, retinal samples taken at known eccentricities were studied from the opposite eye using standard electron microscope methods. RESULTS: Rod opsin was detected at Fd66 in or near the fovea, and a second focus appeared at Fd75 to Fd77 near the optic disc in the nasal rod ring. The earliest opsin appeared in the apical stubs, which resembled the apical connecting cilium in the electron microscope. Staining of the entire cell body membrane, including the synaptic spherule, was present 4 to 7 days later. Opsin expression had a nasal bias with rods at the nasal ora labeled at Fd140, whereas temporal ora was not labeled until Fd155. Cell body labeling disappeared by Fd132 across central retina but persisted into the first postnatal year in far peripheral retina. Outer segment (OS) length measurements showed that rods in the rod ring had the longest OS between Fd115 and postnatal week 9. Rod OS at all retinal eccentricites continued to elongate between 11 months of age and adulthood. CONCLUSIONS: Rod opsin expression follows a foveal-to-peripheral gradient beginning at Fd66 and ending near birth. Rod opsin is detected first in the connecting cilium and slightly later in the entire cell membrane, and then cell membrane labeling disappears as the heavily labeled OS elongates. Although the first OS appear on rods near the fovea, these OS still are short at birth and do not reach adult length until after 2 years of age. The longest OS at birth are found on rods at the rod ring, suggesting that this region could have higher scotopic sensitivity than central retina at birth. PMID- 7499087 TI - Differential temporal and spatial expression of insulin-like growth factor binding protein-2 in developing chick ocular tissues. AB - PURPOSE: To determine the developmental expression and localization of mRNA for insulin-like growth factor binding protein-2 (IGFBP-2), a major binding protein of IGF-I and IGF-II, in ocular tissues of the embryonic and early posthatched chick. METHODS: In situ hybridization and northern blot analysis were used to analyze the cellular origin and relative expression of IGFBP-2 mRNA in ocular tissues. RESULTS: Wholemount in situ hybridization reveals that, as early as 3.5 days of embryonic development (E3.5), IGFBP-2 mRNA is already expressed in many areas of the embryo, including surface ectoderm, certain regions of the brain, pharyngeal clefts, somites, and limb buds. In the eye, IGFBP-2 mRNA is expressed only in the presumptive corneal epithelium at this time. By E6, IGFBP-2 mRNA expression is present in both the corneal epithelium and endothelium. By E12, IGFBP-2 mRNA is detected clearly in the corneal stroma as well as in several other ocular structures, such as the sclera, eyelid, and ciliary body. In the neural retina, a low, diffuse expression of IGFBP-2 mRNA is found at E6, which becomes more localized to the nuclear layers by E12. Northern blot analysis confirms that a high level of IGFBP-2 expression is present in the cornea and sclera by E8 to E12. A high level of IGFBP-2 mRNA expression, however, is not observed in the retina until E18. At posthatch day 2 (P2), northern blot analyses of ocular tissues reveal that the cornea contains the highest ocular level of IGFBP-2 mRNA expression, a value equal to that of brain and liver. CONCLUSIONS: The early appearance, along with differential temporal and spatial expression of IGFBP-2 mRNA in developing ocular tissues, suggests a role for IGFBP-2 in the regulation of growth and differentiation of several ocular tissues, including the cornea, sclera, and retina. PMID- 7499088 TI - Delayed visual evoked potentials in adults after monocular visual deprivation by a dense cataract. AB - PURPOSE: To study the effect of monocular visual deprivation caused by dense unilateral cataracts in adults. METHODS: Visual evoked potentials have been recorded in 11 patients after removal of a dense unilateral cataract acquired in adulthood. These were compared with those from 8 control patients after removal of a mild lens opacity. RESULTS: Visual evoked potentials recorded on the first day after removal of a longstanding, dense, unilateral cataract showed a marked delay to stimulation of the operated eye compared to the unoperated eye. No delay was found in the operated or unoperated eye of control patients. Delays in the visual evoked potential returned to normal within approximately 3 months after surgery, with the exception of the two patients with the most longstanding cataracts in whom the delays persisted much longer. CONCLUSIONS: The adult central visual system is sensitive to visual deprivation caused by a longstanding, dense, unilateral cataract. The changes found may be important in understanding the causes of intractable diplopia, which sometimes occurs after the removal of such cataracts. PMID- 7499089 TI - Refractive error and axial length in a primate model of strabismus and congenital nystagmus. AB - PURPOSE: To evaluate the development of refractive error and axial length in a primate model of sensory strabismus and nystagmus. METHODS: Four macaque monkeys had alternating tarsorraphy beginning within 24 hours of birth. One eye was closed for 25 days, and when it opened, the second eye immediately was closed for the next 25 days. Cycloplegic refractions and axial lengths were determined prospectively for three animals for 1 year or more. These data were compared to those of three unsutured control macaques raised under otherwise similar conditions. RESULTS: Each experimental animal developed exotropia and nystagmus. The first occluded eyes were significantly more hypermetropic than the control eyes at 1 month of age (+7.25 D +/- 1.95 D versus +1.92 D +/- 1.27 D; P < 0.02) and remained significantly more hypermetropic throughout the study. The second occluded eyes were more hypermetropic than the control eyes at 1 month (+2.42 D +/- 2.13 D versus +1.92 D +/- 1.27 D; P = 0.20), but less hypermetropic than the first occluded eyes. The rate of emmetropization was slightly faster for the first occluded eyes than for the control eyes (-0.12 D/month compared to -0.03 D/month). The mean axial length measurements of the experimental and control eyes were the same at 1 month, and their rates of change over time were identical. CONCLUSIONS: Persistent hypermetropia was produced by a brief period of reverse neonatal eyelid closure in a model of congenital-like nystagmus. It is suggested that infantile lid closure, nystagmus, or amblyopia after neonatal visual disruption may be associated with a failure of normal emmetropization. PMID- 7499090 TI - The reliability of a video-enhanced Hirschberg test under clinical conditions. AB - PURPOSE: To examine the reliability and usefulness of a video-based Hirschberg test under clinical conditions. METHODS: The authors estimated ocular deviation in 87 patients with strabismus through automated analysis of corneal reflex displacement using a video refractor. The reproducibility of measurement, the comparison with the prism and alternate cover test (PACT), and the distribution of the Hirschberg ratio were investigated. RESULTS: The 95% limits of agreement of the video-based Hirschberg test evaluated by repeated measurements were +/- 0.18 mm (equivalent to +/- 2.2 degrees or +/- 3.8 prism diopters [PD] of calculated strabismic deviation) for the horizontal deviation and +/- 0.28 mm (equivalent to +/- 3.4 degrees or +/- 5.9 PD) for the vertical deviation. The 95% limits of agreement between the Hirschberg measures and the PACT were within +/- 7.8 degrees or +/- 13.7 PD. The average (+/- SD) Hirschberg ratio was 12.3 +/- 1.2 degrees/mm or 21.8 +/- 2.1 PD/mm. CONCLUSIONS: The video-enhanced Hirschberg measurement shows good reproducibility and ease of application, even in the testing of infants. In quantitative analysis, however, systematic measurement error resulting from intersubject variance of the Hirschberg ratio should be taken into consideration. PMID- 7499091 TI - Conjugacy of eyelid movements in vertical eye saccades. AB - PURPOSE: To examine the conjugacy of lid saccades in normal subjects. METHODS: Saccades of both upper eyelids were recorded simultaneously by means of small, lightweight, magnetic search coils, fixed on each lid. Subjects then made vertical eye saccades between fixed targets. The associated eye saccades were recorded simultaneously by means of magnetic search coils. The authors further examined whether the position of the lid coils affected the recordings. RESULTS: Lid saccades were not as conjugate as their associated eye saccades. Nonconjugacy of lid saccades averaged approximately 8% of the target amplitude, irrespective of saccadic direction. By contrast, nonconjugacy of the associated eye saccades averaged approximately 1% of the target amplitude. Coil position significantly (P < 0.05) affected the recordings. CONCLUSIONS: Lid saccades are not as conjugate as their associated eye saccades. Coil position is critical for the recordings of lid saccades. PMID- 7499092 TI - Accommodation to monochromatic and white-light targets. AB - PURPOSE: The objective of the current study was to compare accommodation to targets illuminated with monochromatic light from different regions of the visible spectrum with accommodation to white-light targets. METHODS: One of 10 marrow-band interference filters (430, 450, 470, 500, 530, 550, 570, 590, 630, and 670 nm) was used to produce monochromatic light from a tungsten-halogen source to illuminate a Maltese cross-target in Maxwellian view. Luminance of each monochromatic light was matched by minimum border photometry against a standard white light (3000 K) that was maintained at 200 cd/m2. Chromatic difference of focus of the eye was minimized for all monochromatic targets by the use of an achromatizing lens. A white-light target also was used, and the subject's eye was achromatized or the eye had normal chromatic aberration. The target was moved sinusoidally toward and away from the eye at a temporal frequency of 0.2 Hz over a 1 D amplitude (peak to peak). Accommodation was monitored continuously by an infrared recording optometer, and responses were Fourier analyzed to obtain gain and phase lag at the temporal frequency of stimulation. RESULTS: Accommodative gain was highest and phase lag was smallest when the target was illuminated by white light in the presence of normal chromatic aberration. The achromatized white-light gain of accommodation was statistically similar to the gain for monochromatic targets, indicating that the presence of chromatic aberration facilitates accommodation. Significant intersubject variability was present in the accommodative tracking ability to monochromatic targets. CONCLUSIONS: Accommodation to monochromatic targets is not as accurate as accommodation to a white-light target, and this effect is related to the presence of ocular longitudinal chromatic aberration for the white-light target. PMID- 7499094 TI - Cell-specific expression of the human Na+,K(+)-ATPase beta 2 subunit isoform in the nonpigmented ciliary epithelium. AB - PURPOSE: To evaluate the patterns of expression of beta subunit isoforms of the Na+,K(+)-ATPase and H+,K(+)-ATPase in the human eye and to determine the cell specific distribution of the beta 2 subunit in the human ciliary epithelium. METHODS: Total RNA extracted from human ocular tissues was screened by Northern blot analysis with cDNA probes for the human Na+,K(+)-ATPase subunit isoforms (beta 1 and beta 2) or the H+,K(+)-ATPase (alpha and beta) subunits. Antibodies were raised to the amino and carboxyl terminal regions of the human beta 2 isoform. Polymerase chain reaction was used to verify the expression of beta 2 subunit in nonpigmented ciliary epithelial cells (NPE). RESULTS: Transcripts for the Na+,K(+)-ATPase beta 1 and beta 2 subunit isoforms were present at different levels in all the ocular tissues except the lens, which expressed only beta 1. No transcripts for the alpha or beta subunits of the H+,K(+)-ATPase were detected in the eye. Isoform beta 2 specific anti-peptide antibodies V15E (N-terminus) and A18R (C-terminus) recognized a 55- to 60-kDa protein in the ciliary epithelium and the core protein of 32 kDa after N-glycanase treatment. Immunocytochemical localization within the ciliary epithelium indicates that the Na+,K(+)-ATPase beta 2 isoform is expressed preferentially in the NPE cells. The expression of Na+,K(+)-ATPase beta 2 isoform in the human NPE cell line, ODM-2, was confirmed by polymerase chain reaction amplification and Southern blot analysis. CONCLUSIONS: The Na+,K(+)-ATPase beta 2 subunit isoform, but not H+,K(+)-ATPase, was expressed widely in ocular tissues of the human eye. The restricted cellular distribution of beta 2 isoform within the NPE cells represents an important differential gene marker associated with the multiple alpha subunit isoforms of Na+,K(+)-ATPase. PMID- 7499093 TI - Characterization of potassium and chloride channels in the basolateral membrane of bovine nonpigmented ciliary epithelial cells. AB - PURPOSE: The pigmented epithelial (PE) and the nonpigmented epithelial (NPE) cells of the ciliary body may function as a syncytium for secretion of the aqueous humor, with solute and water entering through the PE and leaving through the NPE cell. Secretion across the basolateral membrane of the NPE cell was postulated to depend on coupling sodium extrusion through the NaK-ATPase to efflux through Cl channels. In the current study, the authors used single-channel patch clamp techniques to characterize the ion channels present in the basolateral membrane of the NPE cell. METHODS: Mild enzymatic digestion of the bovine ciliary process was used to obtain pairs or clusters of NPE cells that were coupled to their neighbor PE cells. Cells were dispersed immediately onto a coverslip, bathed in a HEPES-buffered saline, and imaged with an inverted microscope. With the anatomic relationship between cells still intact, a patch pipette electrode was applied to the basolateral surface of the NPE cell. Single channel currents were then characterized in 52 cells by using either the cell attached or excised versions of the patch-clamp technique. RESULTS: A calcium dependent "maxi" K channel was found in approximately 33% of the patches. It was activated by depolarizing voltage steps in cell-attached patches and was relatively inactive near the resting potential. When excised from the cell, it was activated by high levels of bath calcium and, in symmetrical K solutions (150 mM), showed a linear current-voltage (I-V) relationship with a slope conductance of approximately 150 pS. This channel was highly selective to K over Na and was blocked by barium (2 mM) or quinine (1 mM). In cell-attached recordings, a smaller conductance Cl channel was observed in 73% of the patches. The channel currents were inward at resting potential and outward with depolarizing voltage steps, with a pipette reversal potential of approximately -23 mV. The current voltage relationship of this channel was nearly linear with a slope conductance of approximately 24 pS. When pipette Na and K were replaced with N-methyl-D glucamine, this channel produced an inward current showing a high selectivity for Cl. This Cl-selective channel was activated by dibutyryl-cyclic adenosine monophosphate (cAMP) but not by elevation of intracellular calcium with ionomycin or by anisosmotic cell swelling. CONCLUSIONS: Two ion channels were found in the NPE basolateral membrane: One was K selective, the other was Cl selective. The cAMP-activated Cl channel may be important in supplying the counterion for Na extrusion across the basolateral surface of the NPE cell, whereas the calcium dependent maxi K channel may be useful for membrane hyperpolarization to increase the driving force for Cl exit. PMID- 7499095 TI - Localization of alpha 2-adrenergic receptor subtypes in the anterior segment of the human eye with selective antibodies. AB - PURPOSE: To develop antibodies that selectively recognize each of the alpha 2 adrenergic receptor (AR) subtypes and to determine the expression and localization of these subtypes in the anterior segment of the human eye. METHODS: Recent studies have shown that there are three subtypes of the alpha 2-ARs, termed alpha 2-C10 (alpha 2A), alpha 2-C2 (alpha 2B), and alpha 2-C4 (alpha 2C). Polymerase chain reaction was used to amplify portions of these receptors fused (in-frame) to a cDNA encoding glutathione-S-transferase (GST). The expressed fusion proteins were used to immunize chickens, and antibodies were generated. Immunofluorescence microscopy was used to localize the alpha 2-AR subtypes in sections of human and rabbit ciliary body. Polymerase chain reaction and dot blot hybridization were used to determine which subtypes were present in RNA from primary cultures of human nonpigmented epithelium (NPE) and rabbit iris-ciliary body (ICB). RESULTS: Immunofluorescence microscopy of COS cells transfected with the alpha 2-AR subtypes showed that the antibodies raised against the GST receptor fusion proteins specifically recognized their respective receptor subtypes. In the human ciliary body, alpha 2 B and alpha 2C immunoreactivity were present in the NPE and ciliary muscle. In the rabbit ciliary body, alpha 2A immunoreactivity also was present. Polymerase chain reaction and dot blot hybridization indicated that RNA encoding the alpha 2B and alpha 2C subtypes was present in human NPE and that RNA encoding all three subtypes was present in the rabbit ICB. CONCLUSIONS: Multiple alpha 2-adrenergic subtypes are expressed in the ciliary body. In the human, alpha 2B and alpha 2C predominate, whereas all three are present in the rabbit. This could be important with respect to animal models of glaucoma and to the development of drugs for lowering intraocular pressure. PMID- 7499097 TI - Local connected fractal dimensions and lacunarity analyses of 60 degrees fluorescein angiograms. AB - PURPOSE: The retinal vascular tree exhibits fractal characteristics. These findings relate to the mechanisms involved in the vascularization process and to the objective morphologic characterization of retinal vessels using fractal analysis. Although normal retinas show uniform patterns of blood vessels, in pathologic retinas with central vein or artery occlusions, the patterns are irregular. Because the generalized box fractal dimension fails to differentiate successfully between normal and abnormal retinal vessels in 60 degrees fluorescein angiograms, the authors have further investigated this problem using the local connected fractal dimension (alpha). METHODS: The authors studied 24 digitized 60 degrees fluorescein angiograms of patients with normal retinas and 5 angiograms of patients with central retinal vein or artery occlusion. The pointwise method estimated the local complexity of the angiogram within a finite window centered on those pixels that belong to the retinal vessels. Color-coded dimensional images of the angiograms were constructed by plotting the pixels forming the object with a color that corresponded to specific values of alpha +/- delta alpha. RESULTS: The color-coded representation allowed recognition of areas with increased or decreased local angiogram complexity. The alpha distributions showed differences between normal and pathologic retinas, which overcomes problems encountered when using the methods of calculating the generalized fractal dimensions. A multivariate linear discriminant function using parameters from the alpha distribution and a further fractal parameter--lacunarity- reclassified 23 of the 24 normal and 4 of the 5 pathologic angiograms in their original groups (total: 92.1% correct). CONCLUSIONS: This methodology may be used for automatic detection and objective characterization of local retinal vessel abnormalities. PMID- 7499096 TI - Studies of adenovirus-induced eye disease in the rabbit model. AB - PURPOSE: To achieve a better understanding of the pathogenic processes associated with human adenovirus (Ad)-induced ocular disease. METHODS: Growth curves of Ad5 and Ad14 were performed in cell cultures derived from rabbit and human corneal epithelium (CE) and corneal keratocytes (CK). For in vivo studies, rabbit eyes were inoculated intrastromally and topically with 10(6) plaque-forming units per eye of Ad5 and ultraviolet light-inactivated (UV-1) Ad5 or Ad14, and the clinical features of the eyes were evaluated by biomicroscopic slit lamp examinations. Duration and quantitation of virus in tear samples were monitored. Humoral response was evaluated by enzyme-linked immunosorbent assay and serum neutralization titrations. Histopathologic and immunocytochemical staining of frozen corneal tissues was performed to determine the expression of major histocompatibility complex (MHC) class I and II and the presence of CD4+ and CD8+ T lymphocytes and CD18+ cells after the immunopathologic response elicited by virus inoculation. RESULTS: Both Ad5 and Ad14 replicated in all human cell cultures studied. In cells of rabbit origin, Ad5 replicated in cultured CE and CK cells, whereas Ad14 replication appeared restricted. Virus titers in ocular samples from Ad5-inoculated eyes peaked on postinoculation days 3 through 4, with approximately a 100-fold increase in infectious virus in comparison to initial titers. The duration of Ad5 shedding was 8.9 +/- 2.4 days. Ad5, Ad5 UV-I, and Ad14 induced seroconversion and subepithelial opacities. CD4+ and CD8+ T lymphocytes and CD18+ cells were present in these intrastromal immune cell infiltrates. Expression of MHC class I and II was observed in keratocytes and immune cells; MHC class I also was expressed on CE cells in inflamed areas. CONCLUSIONS: Ad5 is capable of replicating in both CE and CK cells of the rabbit eye. The presence of Ad antigens within the corneal stroma originating from infectious virus (Ad5), UV-inactivated virus (Ad5), or nonreplicating infectious virus (Ad14) can elicit indistinguishable immunopathologic responses in the stroma composed of CD4+, CD8+, and CD18+ cells. PMID- 7499098 TI - Measurement of carotenoids, retinoids, and tocopherols in human lenses. AB - PURPOSE: To determine the levels of carotenoids, retinoids, and tocopherols in normal and cataractous human lenses. METHODS: Concentrations of carotenoids, retinoids, and tocopherols were measured by high-performance liquid chromatography (HPLC) in 12 American normal lenses, 9 American cataractous lenses, and 10 Indian cataractous lenses. RESULTS: On a per gram wet weight of tissue basis, human lenses contained 11 to 25 ng xanthophylls (lutein and zeaxanthin), 31 to 50 ng retinol, 21 to 25 ng retinyl palmitate, 1573 to 2550 ng alpha-tocopherol, and 257 to 501 ng gamma-tocopherol. Concentrations of lutein, zeaxanthin, and retinol were significantly higher in Indian cataractous lenses than in American normal or cataractous lenses. There were no differences in the lutein-zeaxanthin, retinoid, or alpha-tocopherol contents between American normal lenses and American cataractous lenses. The range of ratios of lutein to zeaxanthin in human lenses was 1.6 to 2.2. The mean age of the Indian lens donors was 20 years younger than the American lens donors. Comparisons using contralateral lenses indicated that there was significant interindividual variance in human lens concentrations of xanthophylls, retinoids, and tocopherols. beta-carotene and lycopene, major carotenoids in human serum and other tissues, were not detected in human lenses. CONCLUSIONS: Xanthophylls (lutein and zeaxanthin) are the only carotenoids detected in human lens. Retinol, retinyl palmitate, and alpha- and gamma-tocopherols also are present in human lens. Determinants of lens concentration of nutrients are not defined, but dietary factors are likely to be important. PMID- 7499099 TI - Comparative analysis of proliferating cell nuclear antigen, bromodeoxyuridine, and mitotic index in uveal melanoma. AB - PURPOSE: Recent production of a monoclonal antibody, PC10, against proliferating cell nuclear antigen (PCNA) makes it possible to evaluate cell cycling in formalin-fixed tissues. In this study, the authors quantitatively evaluated the relationship between PCNA expression and two other measures of cell cycling, bromodeoxyuridine labeling index (BrdU LI) and mitotic index (MI), in archival uveal melanomas. The authors also examined the relative prognostic importance of each measure. METHODS: Serial sections from 35 formalin-fixed, paraffin-embedded uveal melanomas were immunostained with PC10 and BrdU antibody using a standard avidin-biotin-peroxidase method. A quantitative scoring system was used to evaluate the fraction of cells that were positive for PCNA, BrdU, and mitotic figures in the regions of high cycling. The LIs of the different markers were compared, and their prognostic importance was evaluated. RESULTS: The median PCNA LI was 3.05% compared to the median BrdU LI of 0.94% and the median MI of 0.034%. The PCNA LI was more variable in replicate sections than either the MI or the BrdU LI. The correlation between PCNA LI and BrdU LI was 0.58, between PCNA LI and MI it was 0.46, and between BrdU LI and MI it was 0.81. The relative risk of tumor-related mortality per doubling of BrdU LI was 2.35, and of MI it was 2.34. Although these were significant, PCNA LI of 1.08 was not. CONCLUSIONS: Proliferating cell nuclear antigen immunostaining did not demonstrate a strong relationship with either BrdU LI or MI. Unlike MI and BrdU LI, PCNA LI was not correlated with tumor-related mortality. Caution is warranted in the interpretation of PCNA immunostaining in uveal melanomas. PMID- 7499100 TI - Spasmus nutans. A long-term follow-up. AB - PURPOSE: Nystagmus, head nodding, and anomalous head position are symptoms of spasmus nutans. This disorder appears in early childhood and is thought to be self-limited. However, the visual outcome of patients with spasmus nutans is unclear. The resolution of nystagmus has not been proven with quantitative eye movement recordings. The purpose of this study was to perform long-term follow-up examinations (mean, 5.5 years) of patients with spasmus nutans. METHODS: Ten patients with spasmus nutans were followed up clinically until a mean age of 7 years. Included were quantitative eye and head movement recordings. RESULTS: At their last examination (mean age, 7.1 years), visual acuity in four patients was 20/20 in both eyes, in five patients it was 20/30 or better in at least one eye, and in one patient it was 20/50 in each eye. Three patients had orthotropia with normal stereo acuity. The remaining patients had esotropia, dissociated vertical deviation, amblyopia, or latent nystagmus. All patients had fine, intermittent asymmetric, pendular nystagmus on eye movement recordings. CONCLUSIONS: Good visual acuity can be expected in patients with spasmus nutans; one third have normal stereo acuity. However, subclinical nystagmus persists until at least 5 to 12 years of age. PMID- 7499101 TI - mRNA from frog corneal epithelium increases water permeability in Xenopus oocytes. AB - PURPOSE: To investigate the existence of a water channel in the frog corneal epithelium by studying the osmotic water permeability (Pf) of Xenopus oocytes expressing the mRNA message from frog corneal epithelium. METHODS: Total RNA was obtained from corneal epithelium by a single-step phase separation method, and poly A+ RNA was isolated using oligo-dT columns. This mRNA was injected into the oocytes. After a 48-hour incubation, oocyte volume changes elicited by a hypoosmotic solution were measured with a computerized video system. RESULTS: Oocytes injected with 50 nl mRNA (1 microgram/microliter) showed a significant increase in Pf compared to water-injected controls (8.4 +/- 1.5 to 17.5 +/- 1.9 cm.sec-1 x 10(-4), P < 0.005). mRNA-injected oocytes exposed to a higher external [Cl-] showed a heightened permeability. Furthermore, Pf of oocytes exposed to a solution containing the recognized water-channel blocker HgCl2 was significantly lower than the Pf of mRNA-injected oocytes not exposed to HgCl2. CONCLUSIONS: Evidence was found for a water channel in the frog corneal epithelium because oocytes injected with the epithelial mRNA manifested increased water permeability. The increase in water permeability was larger in the presence of external Cl- and was inhibited by HgCl2. This finding correlates with measurements of Pf in the intact epithelium in which apical Cl- induced an increase in transepithelial water permeability prevented by HgCl2. PMID- 7499102 TI - Response of human tumor xenografts in athymic nude mice to docetaxel (RP 56976, Taxotere). AB - Docetaxel (Taxotere, RP 56976, NSC 628503), a new taxoid, was evaluated for preclinical evidence of anticancer activity in athymic nude (NCr-nu) mice bearing established, subcutaneously (s.c.) implanted human tumor xenografts CX-1 or KM20L2 (colon carcinomas), LX-1 (lung carcinoma), MX-1 (mammary carcinoma), and SK-MEL-2 (melanoma). Other evaluations used OVCAR-3 (ovarian carcinoma) xenografts implanted intraperitoneally (i.p.). Docetaxel was administered intravenously (i.v.) every 4 days for 3 injections (q4d x 3) except for one OVCAR 3 experiment in which the drug was given i.p. every 7 days for 3 injections. Tumor measurements, animal body weights, and mortality were determined. The highest dosage used (50 mg/kg/dose) was toxic in all experiments in which the 4 day treatment interval was used. The maximally tolerated dosage (MTD) ranged from 15 to 33 mg/kg/dose. Therapeutic responses among these xenografts ranged from clinically important long-term tumor-free survivors (MX-1, SK-MEL-2, and OVCAR-3) to tumor growth delays of various durations (CX-1, LX-1, and KM20L2). The response of SK-MEL-2, a xenograft highly refractory to available drugs, was particularly noteworthy. These results are indicative of a broad spectrum of antitumor activity for docetaxel. PMID- 7499103 TI - P-glycoprotein mediated resistance to 5'-nor-anhydro-vinblastine (Navelbine). AB - Navelbine (NVB, vinorelbine tartrate) is a semisynthetic Vinca alkaloid in which the catharanthine moiety contains an eight-membered ring in place of the nine membered ring that is present in all naturally occurring members of the vinblastine group. This modification selectively reduces interaction with anoxal vs mititotic microtubules and may account for the lower neurotoxicity with improved antitumor activity that has been observed in clinical trials with breast, lung and ovarian cancer. We were interested in whether the structural modification in NVB would also alter the drug resistance profile. Specifically, our aim was to determine whether NVB, like vinblastine (VBL), participates in P glycoprotein (P-gp)-mediated multidrug resistance (MDR). NVB-resistant, murine P388 cells (P388/NVB), were derived in vivo and used in conjunction with a battery of drug-resistant P388 cell lines in vivo and murine and human tumor cell lines in vitro to develop a resistance profile for NVB. P388/NVB bells were cross resistant to drugs involved in MDR (doxorubicin, etoposide, amsacrine, vinblastine, vincristine and actinomycin D), but not to the alkylating agents, cyclophosphamide, carmustine, and cisplatin, or to the antimetabolites, 5 fluorouracil and methotrexate. P388/NVB cellular resistance to NVB was stable without drug pressure during continuous passage in vivo for more than ten weeks and in vitro for at least five weeks. These cells exhibited increased expression of P-gp, and a 30-fold level of resistance of NVB in vitro, which was completely reversable with verapamil. The MDR phenotype was confirmed in other tumor models. P388 tumors resistant to vinblastine, vincristine, doxorubicin, and etoposide were cross-resistant to NVB in vivo.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7499105 TI - Hepsulfam distribution in blood, plasma and cerebrospinal fluid of baboons. AB - The alkylating agent Hepsulfam (Sulfamic acid 1,7-heptanediyl ester, NSC 329680) was developed as a more hydrophilic analog of busulfan. The objective of this study was to determine partitioning of hepsulfam between blood, plasma, and cerebrospinal fluid (CSF) in two female baboons following intravenous administration. Hepsulfam was administered at 11 mg/kg, and blood and CSF levels were determined by gas chromatography with electron capture detection. Blood levels were fairly constant between animals (17-25 and 20-23 micrograms/ml) for six hours after administration, following peak levels of 43 and 33 micrograms/ml, respectively, for the two animals. Peak plasma levels of 35 and 36 micrograms/ml were achieved, and initial plasma half-lives in baboons were similar to those seen in other species, with a t1/2 alpha of 1 h. The plasma terminal half life of 0.2 h, estimated from limited sampling times, was shorter in baboons than in mice, dogs, or humans. Baboon CSF levels decreased from 1.7 to 0.3 micrograms/ml during 6 h post infusion, and peak concentrations in CSF lagged behind plasma levels. CSF/plasma ratios ranged from 0.33 to 0.62 in one animal, whereas ratios of 0.2-0.25 were maintained in the other animal during the same period. Results from this study indicate hepsulfam will enter the CSF following intravenous administration, and the CSF/plasma ratios are lower than those obtained following oral busulfan administration. PMID- 7499104 TI - Preclinical antitumor activity of ethyldeshydroxysparsomycin in combination with cisplatin. AB - The efficacy of cisplatin (CDDP) in combination with the protein synthesis inhibitor ethyldeshydroxysparsomycin (EDSM) has been tested in two tumor models at various schedules. Mice with L1210 leukemia or B16 melanoma were treated with CDDP alone or in combination with EDSM. Against L1210 leukemia, which is sensitive to CDDP, combinations elicited increases in life-span for all treatment schedules compared to those achieved with the corresponding dose of CDDP. Moreover, the combination of EDSM with this platinum compound yielded a cure rate > 80%, compared to < 35% for single CDDP treatment. Although the B16 melanoma is rather resistant to both CDDP and EDSM, combinations of these agents against B16 melanoma showed schedule dependent efficacy and in certain schedules significant therapeutic advantage over individual drug treatment, but cures were not observed. Our results suggest that EDSM has significant synergistic capabilities in both animal tumor models, but strong therapeutic enhancement of cisplatin efficacy is only seen when the tumor is sensitive to CDDP. PMID- 7499106 TI - Binding of a new multidrug resistance modulator, S9788, to human plasma proteins and erythrocytes. AB - The interactions of S9788 with human plasma proteins have been investigated in vitro by an erythrocyte partitioning technique that allows an estimation of the plasma proteins and erythrocytes binding parameters. S9788 was 98% bound to plasma and blood. Lipoproteins bound S9788 with high affinities (binding constants of 0.645, 12.8 and 87.0 x 10(6) M-1 for HDL, LDL and VLDL, respectively) and accounted for more than 55% of the total circulating S9788. Albumin and alpha 1-acid glycoprotein also bound S9788 with lower binding constants of 0.022 and 0.245 x 10(6) M-1. S9788 was mainly distributed in the plasma blood compartment (75-80%) with blood-to-plasma concentrations ratio of 0.6 to 0.7. These results indicate that, in vivo, the fraction of blood S9788 available for tissue diffusion, i.e., the free drug fraction in blood, should depend on lipoprotein concentration in plasma. PMID- 7499107 TI - Adenocarcinoma of unknown primary: retrospective analysis of chemosensitivity of 313 freshly explanted tumors in a tumor cloning system. AB - Cancer of unknown primary origin is the eighth most common form of malignancy and accounts for up to 10% of all neoplasms diagnosed. It is a set of heterogenous tumors with widely varying sensitivity to systemic chemotherapy. Over the past years progress has been made in identifying subsets of patients that can be effectively treated with chemotherapy and may achieve long-term survival even with metastatic disease. However, the large majority of cancers of unknown origin still is resistant to chemotherapy. In an attempt to identify conventional and investigational new agents with possible activity against cancers of unknown primary, we have retrospectively analyzed the results of chemosensitivity testing in a soft agar cloning system in vitro and have compared these data with published clinical trials. Between 1978 and 1993, a total of 19584 tumor specimens were studied using a variety of investigational or established antitumor agents. Of these, 615 (3.1%) were tumors of unknown origin and confirmed on pathology review. The largest histologic subgroup was adenocarcinoma (332, 54%). Sufficient numbers of cells for in vitro testing were obtained from 313 tumor specimens (94.3%). Of 278 agents tested in adenocarcinoma of unknown origin, borderline activity (< 20% in vitro response) was noted for 5-FU, doxorubicin, bleomycin, mitoxantrone, mitomycin-C, cisplatin, and etoposide. In vitro response rates of > or = 20% were observed for actinomycin-D, BCNU, melphalan, methotrexate, taxol, and vinblastine. In addition, several investigational agents including fludarabine, amira235, bisantrene, Dupont840, echinomycin, tiazofurin, LY104208 (vinzolidine), intoplicine, and topotecan had activity.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7499108 TI - Stimulation of tumor growth in vitro and in vivo by suramin on the VX2 model. AB - Suramin is an antitrypanosomal compound with confirmed efficacy against several human malignancies. It is generally assumed that its mechanism of action includes the interaction with different growth factors, unlike most of the anticancer drugs. Its anticancer activity has not been tested in vivo against squamous cell carcinoma. The purpose of this study was to assess the efficacy and toxicity of suramin in vivo and in vitro on the VX2 tumor model at therapeutic monitored plasma concentrations. We determined the pharmacokinetics of suramin in rabbits, and modelized its administration in order to obtain plasma concentrations between 150 and 300 micrograms/ml throughout the treatment course of 3 weeks. Under these conditions, antitumor effects of suramin were evaluated in vivo by comparing liver tumor involvement in suramin-treated and control rabbits. Liver involvement was quantified by image analysis and in vitro effects were also determined at the same concentrations. In vivo, suramin promoted liver tumor growth significantly (p < 0.05), compared to untreated controls. In vitro, suramin significantly stimulated tumor cell growth at concentrations above 200 micrograms/ml (p < 0.01). Suramin may have stimulatory effects on tumor growth in squamous cell carcinoma at relevant plasma drug concentrations. Caution should be taken in further trials in patients with squamous cell carcinomas. PMID- 7499109 TI - Phase I trial of pentosan polysulfate. AB - Pentosan polysulfate is a semisynthetic pentasaccharide heparinoid derived from beechwood shavings. A total of nineteen patients with various adult solid tumors were treated with three dose levels (15, 22.5, and 30 mg/m2/dose) of subcutaneous pentosan polysulfate every 6 hours. The dose limiting toxicities were thrombocytopenia and elevated transaminases at the dose of 30 mg/m2 every 6 hours. The recommended starting dose for phase II trials is 22.5 mg/m2 given every 6 hours. There was an increase in anticoagulant activity as measured by activated partial thromboplastin time (aPTT) at the dose of 22.5 mg/m2 every 6 hours in most patients. There were no objective responses and three patients had stable disease lasting 16, 19 and 76 weeks. PMID- 7499111 TI - Phase II trial and cost analysis of fazarabine in advanced non-small cell carcinoma of the lung: a Southwest Oncology Group study. AB - BACKGROUND: Fazarabine is a novel nucleoside with broad spectrum pre-clinical activity and was chosen for study in patients with incurable non-small cell carcinoma of the lung. The expenses associated with investigational treatment have been assumed to be more than what would occur with conventional therapy, however, data are limited. METHODS: Twenty-three patients with metastatic non small cell lung cancer were treated with fazarabine. Fazarabine was administered as a 72 hour continuous infusion at 2.0 mg/M2/hour. A cost analysis of treatment was calculated for patients treated in Springfield, MO. RESULTS: There were no responses (0%, 95% confidence interval = 0-15%) and median survival was 8 months. An analysis of the cost of treatment in the 4 patients treated in Springfield, MO, was compared to the costs of treatment with 4 cycles of cisplatinum and etoposide. There were no significant differences in costs for patients treated with the investigational agent as compared with conventional chemotherapy. CONCLUSIONS: Fazarabine has no demonstrable activity in patients with metastatic non-small cell carcinoma of the lung. Treatment with this agent in an investigational setting was no more expensive than treatment with conventional chemotherapy. PMID- 7499112 TI - Phase II study of cisplatin, 5-fluorouracil and interferon-alpha in recurrent carcinoma of the cervix. AB - Standard cytotoxic regimens have failed to prolong survival in patients with recurrent cervical cancer; therefore, new agents or combinations of agents are necessary to improve outcome. Cisplatin and 5-fluorouracil are one of the most active regimens currently available for recurrent cervical cancer. Interferon alpha and potentiate the in vitro and in vivo activity of both agents. Therefore, we sought to define the feasibility and activity of adding interferon-alpha to the cisplatin-5-fluorouracil regimen. Patients with histologically proven recurrent cervical carcinoma received interferon-alpha (5 x 10(6) units/m2/day for 5 days) combined with cisplatin (75 mg/m2 on day 1) plus 5-fluorouracil (500 mg/m2/day for 5 days) in recurrent cervical cancer. Twenty-six patients were included in this study and received a total of 95 courses of therapy. All 26 had prior radiotherapy and five had prior chemotherapy. This regimen was relatively well tolerated as toxicity was comparable to that of cisplatin plus 5 fluorouracil alone. Major response occurred in 8 patients (31%) and 5 (19%) had complete responses. The median response duration was 6 months (range, 2-34 months) and the median survival duration was 9 months (range, 2-38 months). The addition of interferon-alpha to cisplatin plus 5-fluorouracil is feasible. Further studies will be necessary to determine if the addition of interferon alpha to this regimen is superior to cisplatin plus 5-fluorouracil alone. PMID- 7499110 TI - A phase I study of sulofenur in refractory pediatric malignant solid tumors. AB - The diarylsulfonylureas have shown promise in xenograft models of childhood cancer. Sulofenur has been evaluated in phase I and II trials in adults with a variety of solid tumors, but the toxicity and maximum tolerated dose of sulofenur in children and adolescents have not been determined. In a phase I study, sulofenur was administered to 13 patients with refractory pediatric malignant solid tumors. Daily dosages of 640, 800, and 960 mg/M2 in two divided oral doses were given for 5 consecutive days each week for 3 weeks. The primary and dose limiting toxicity was methemoglobinemia, which occurred at all dose levels and required transfusions of packed red blood cells, administration of methylene blue, or both. Anemia and, less frequently, leukopenia and thrombocytopenia were also observed. A maximum tolerated daily dosage was not defined, as methemoglobinemia was noted with each dosage level. There were no measurable tumor responses. The toxicity of this agent makes it unattractive for further investigation in pediatric patients. PMID- 7499115 TI - Experience of the treatment with Sehydrin (Hydrazine Sulfate, HS) in the advanced cancer patients. AB - The results of Sehydrin (Hydrazine Sulfate, HS) treatment of 740 patients with the advanced, recurrent or metastatic solid tumours of various localizations or malignant lymphomas, for whom all the methods of specific treatment (surgery, radiation, chemotherapy) had been exhausted are presented in this work. The objective response, symptomatic therapeutic effects and toxicity were estimated. Clinically significant objective responses were registered in patients with the soft tissue sarcomas, including neuroblastomas, and paradoxically--in such semimalignant tumours as desmoids. Although the objective response in patients with the lung cancer (90%--non-small cell) was only 4%, stabilization of long duration was registered in 22% of cases connected with the impressive relief of heavy common symptoms in 38.5% of the treated patients. Such a subjective response was established in 46.6% of all the 740 cases. The drug given per os was well tolerated by patients in primary and subsequent courses and did not induce myelosuppression or other significant side effects. On the basis of observations available, Sehydrin may be assessed as an alternative drug for the treatment and symptomatic therapy of patients with some advanced solid tumours and malignant lymphomas at a disease stage when the other methods of treatment can not be used. A possible mechanism of antitumour and symptomatic action is being discussed. PMID- 7499113 TI - A phase II trial of doxorubicin HCl Liposome Injection in patients with advanced pancreatic adenocarcinoma. AB - We conducted a phase II study with doxorubicin encapsulated in a liposomal preparation (Doxorubicin HCl Liposome Injection) at a dose of 75 mg/m2 administered as a one hour infusion once every three weeks. Sixteen patients were entered to the trial and 15 are evaluable for response. Hematologic toxicity was significant. The median white blood count was 1.6 x 10(3)/microliters (range: 0.1 9.5), median absolute neutrophil count 0.6 x 10(3)/microliters (range: 0-5.8) and median platelet count was 142.0 x 10(3)/microliters (range: 20-327). Gastrointestinal toxicity was generally mild. Despite two minor responses in liver metastases with a significant decrease in CEA in one of these patients, no major responses were observed, excluding with 95% confidence, a response rate in excess of 20%. PMID- 7499116 TI - Pharmacology, relative bioavailability, and toxicity of three different oral cyclophosphamide preparations in a randomized, cross-over study. AB - Thirty-six patients were entered on this study to determine the pharmacology, bioavailability, and toxicity of three different oral formulations of cyclophosphamide (Cytoxan, Endoxan, and an investigational direct compression tablet). Patients were randomized with respect to the order in which they received the different oral cyclophosphamide preparations, and received each one for two weeks followed by a two week washout period. Concurrent chemotherapy was allowed provided it remained constant across all 3 courses of cyclophosphamide. Plasma concentrations of cyclophosphamide and phosphoramide mustard were measured by gas chromatography with electron capture detection. Peak plasma cyclophosphamide concentrations and times to peak plasma cyclophosphamide and phosphoramide mustard preparations were significantly greater for Endoxan than for Cytoxan and the investigational direct compression tablet. Drug area under the concentration-time curve (AUC), bioavailability, and plasma elimination half life could not be reliably calculated for Endoxan but were similar for Cytoxan and the investigational formulation. Based on AUC comparisons, bioavailability of parent compound (relative to an oral cyclophosphamide solution) was 85% for Cytoxan and 69% for the investigational formulation. This difference was not significant. There were no significant differences between the 3 formulations with respect to any individual type of toxicity, although the investigational formulation tended to be associated with somewhat less overall toxicity (p = 0.08). PMID- 7499117 TI - Remodeling as metaphor: sex offenders' perceptions of the treatment process. AB - The perceptions of 43 adult male sex offenders were examined to generate a grounded theory of the sexual abuse treatment process. Methodology included interviews, direct observations, and record analysis. Constant comparative analysis was used to collect and analyze data concurrently. The offenders use remodeling as a metaphor for their treatment process; they remodel themselves, their relationships, and their environment, but the foundations of offending are ever present. The remodeling process involves falling apart, taking on, tearing out, rebuilding, doing the upkeep, and moving on. Remodeling as a metaphor offers a concrete, visual, individualized, and safe way to address the treatment process of sex offenders. PMID- 7499114 TI - A phase II trial of piroxantrone in disseminated malignant melanoma. A Southwest Oncology Group study. AB - Piroxantrone is one of the anthrapyrazoles developed in an effort to combine the broad antitumor activity of the anthracyclines with decreased myocardial toxicity. It has shown activity in metastatic melanoma during phase I trials. The Southwest Oncology Group (SWOG) conducted a phase II trial in disseminated malignant melanoma with piroxantrone administered at 150 mg/m2 intravenously over 1 h every 21 days, based upon the phase I experience. Forty-six eligible patients were registered to the trial and 44 were evaluable for response. Two partial responses, Wayne of 6 and 9 months duration were observed for an overall response rate of 5% (95% confidence interval 1%-15%). Thirty-six of 46 eligible patients have died with an estimated median survival of 5 months (95% confidence interval 3-8 months). Toxicities were tolerable with granulocytopenia being the predominant toxicity. Based upon the observed response rate, it is concluded that piroxantrone administered at this dose and schedule has detectable but minimal activity, and does not warrant further investigation in this disease. PMID- 7499118 TI - Modeling the client's world through brief solution-focused therapy. AB - The shift to a managed care philosophy means that clients with psychiatric needs are likely to have both fewer and briefer inpatient hospitalizations. Identification of focused goals and measurable outcomes may not translate easily into inpatient programs that have been more process oriented or have based definitions of improvement on complete or near-complete remission of the admission diagnosis. In the context of these shifts, nursing's focus on maintaining a safe environment, developing a therapeutic relationship, providing information, and valuing holistic and individualized care may be lost in programs that treat all clients in the same "packaged" programs. This article describes the philosophical assumptions that underlie the brief inpatient program. The next article in this issue describes how solution-focused therapeutic modalities are used in the continuum of care. PMID- 7499120 TI - Caregiver mental health and family health outcomes following critical hospitalization of a child. AB - Changes in caregiver and family health and in family health behavior patterns were examined 9 weeks after the hospitalization of a child for an acute, first time illness in a pediatric intensive care unit (PICU). Family and caregiver health included reports of physical, mental, role, and social functioning as well as perceptions of health, pain, and changes in health behavior patterns at 3-7 days after admission to PICU and 9 weeks later. A convenience sample of 20 primary caregivers (all mothers) of children aged 2 days to 17 years served as subjects for the study. Results showed a decrease in mental health scores of all subjects over the 9-week period. Further, those caregivers whose child was rated as having a greater potential for chronicity had a significantly greater decline and were in the range of poor mental health at the 9-week posttest. Seventy percent of subjects reported new health problems in the family posthospitalization, and 43% reported at least one change in family health behavior patterns. Implications for testing the efficacy of mental health nursing consultation and case management during and after critical childhood illness are discussed. PMID- 7499119 TI - Brief inpatient psychiatric treatment: finding solutions. AB - The shift to a managed care philosophy means that clients with psychiatric needs are likely to have both fewer and briefer inpatient hospitalizations. Identification of focused goals and measurable outcomes may not translate easily into inpatient programs that have been more process oriented or have based definitions of improvement on complete or near-complete remission of the admission diagnosis. In the context of these shifts, nursing's focus on maintaining a safe environment, developing a therapeutic relationship, providing information, and valuing holistic and individualized care may be lost in programs that treat all clients in the same "packaged" programs. The previous article in this issue described the philosophical assumptions that underlie our brief inpatient program. This article describes how solution-focused therapeutic modalities are used in our continuum of care. PMID- 7499125 TI - [Rosacea and Helicobacter pylori]. PMID- 7499124 TI - A new perspective on loneliness in later life. AB - Loneliness in later life remains a serious problem despite extensive research across the life span. Unfortunately, most studies of loneliness that include older adults focus on individuals who are already lonely; findings suggest interventions dependent upon external factors such as socialization and functional status. Such interventions are not always feasible for older adults, who may have experienced social and functional losses. Nurses must adopt a new perspective in examining loneliness in later life. This new perspective should include a more positive approach that focuses on older adults who are not lonely even though they may have experienced decreased socialization or physical function. Identification of strategies employed by these older individuals may be used to help many elders avoid loneliness and help others cope with the related losses so frequent in later life. PMID- 7499121 TI - Building interdisciplinary mental health services research teams: a case example. AB - The experience of developing an interdisciplinary research team to study outcomes of psychiatric nurse labor use in free-standing psychiatric hospitals is presented. The creation of roles for research team members, the development of working relationships, and the evolution of additional research projects are discussed. Useful suggestions of strategies for developing research teams are presented within the context of proposal development, successful funding, and implementation of a research study. PMID- 7499122 TI - Historical concept analysis of empathy. AB - An analysis of the concept empathy was performed on archival, narrative accounts of Civil War nursing care using concept analysis strategies. Archival model cases are presented with elucidation of critical attributes, antecedents and consequences, and empirical referents of the concept. The antecedent phenomena of patient cues and consequent phenomena of patient responses serve to clarify instances of nurse empathy and establish preliminary and terminal boundaries of the concept. Empirical referents of the critical attributes identification, introjection, and intervention further explicate this abstract construct. Analysis of contrary, related, and borderline cases extracted from the historical accounts serve to clarify empathy. Most notable to the analysis is the existence of nursing interventions as a critical attribute and the nature of patient responses. Preliminary examination of the model cases indicates patient outcomes improve when empathy is extant in the interaction. PMID- 7499123 TI - Self-concept and woman abuse: a rural and cultural perspective. AB - The prevalence of woman abuse has serious community health ramifications because of its associated psychological responses. Theoretical frameworks have described behaviors arising from interpersonal interactions as being affected by self concept. Therefore, assessing self-concept is critical in research and practice for understanding the behavior of abused women and the effects of self-concept on the physical self. This research arose from clinical practice in a rural Mexican American and non-Hispanic white community. The practice focused on individual and group counseling for women and adolescents in abusive relationships. This study was conducted to refine the theoretical basis for interventions related to self concept and woman abuse for rural Mexican American and non-Hispanic white women. PMID- 7499126 TI - [Interleukin 7 in dermatology. Molecular, immunologic and preclinical aspects]. AB - The vast body of knowledge on interleukin-7 (IL-7) that has accumulated over the last few years, i.e. since its discovery in 1988, is of increasing relevance for dermatologists. Because of its particular immunological effects, IL-7 is of particular interest in two areas of dermatology. First, IL-7 can be a valuable addition to immunotherapeutic approaches to the treatment of malignant melanoma. A first pilot study of IL-7 gene-modified autologous tumour cells in the treatment of metastatic melanoma is therefore currently being initiated by the authors. Then, keratinocytes are efficient IL-7-secreting cells and there is evidence that IL-7 can modulate the presence and proliferation of normal and malignant lymphopoietic cells in the dermis. In addition, IL-7 is presently the most potent growth factor for in vitro cultivation of Sezary cells and for the establishment of Sezary cell lines. This report reviews molecular, immunological and preclinical properties of IL-7, with special reference to the possible therapeutic efficacy of this cytokine in dermatology. PMID- 7499127 TI - [The Malignant Melanoma Central Register of the German Society of Dermatology 1983-1993. Epidemiologic developments and current therapeutic management of malignant melanoma of the skin]. AB - The Central Malignant Melanoma Registry of the German Dermatological Society was founded in 1983 and has meanwhile developed into a major continuously updated multicentre project. Up to June 1994, 19,250 reports of cutaneous melanoma had been received, from 41 departments of dermatology in the former Federal Republic of Germany, from 14 departments in the former German Democratic Republic, from 2 departments in Austria, and from 1 department in Switzerland. Analysis of the data revealed some epidemiological trends over time during the years 1983 to 1993. (1) During the last 10 years the percentage of male patients has steadily increased, from an average of 38% in the year 1983 to 46% in 1993. (2) Early diagnosis of malignant melanoma improved during the period of time investigated. The percentage of diagnoses of primary tumour alone increased between 1983 and 1993. The mean tumour thickness (Breslow) decreased in the West Germany from 1.8 mm to 1.3 mm and in East Germany from 2.5 mm to 1.7 mm. The proportion of nodular melanoma decreased correspondingly from 29% to 14% in the former Federal Republic of Germany and from 40.6% to 22.6% in the former Germany Democratic Republic. During the years 1990 and 1993, 64% of melanoma patients with the primary tumour alone were operated on in two consecutive sessions in the former Federal Republic of Germany and 34.2% of those in the former Germany Democratic Republic. During this period 73.7% of all melanoma patients were operated on under local anaesthesia. In recent years surgical operations were more often performed in two consecutive sessions, mostly under local anaesthesia and with decreasing safety margins, in keeping with the decrease in tumour thickness. The present analysis shows that the Central Malignant Melanoma Registry is an important instrument for investigating trends in clinical epidemiology and treatment of malignant melanoma in the German-speaking countries. PMID- 7499128 TI - [Evaluating polymerase chain reaction for detection of C. trachomatis in urogenital smears]. AB - A total of 222 urogenital specimens were investigated with a commercially available polymerase chain reaction (Amplicor test) for the direct detection of Chlamydia trachomatis. The results were compared with those yielded by the conventional cell culture technique. Using cell culture a urogenital C. trachomatis infection could be detected in 72 of 222 patients. The Amplicor test yielded a positive result in 83. Referred to the detection rate of the Amplicor test and that of the cell culture, sensitivity was 91.2% for the test sensitivity and 79.1% for the cell culture. The specificity of both techniques was 100% when the specimens for which neither both nor either one of the tests gave positive results were considered. In accordance with other studies, this study suggests that tests based on nucleic acid amplification will supersede cell cultures as the gold standard for the detection of C. trachomatis and also become the method of choice in routine diagnosis of urogenital chamydial infections. PMID- 7499129 TI - [Low dosage cyclosporin A therapy in pyoderma gangrenosum. Experiences with 6 patients]. AB - Pyoderma gangraenosum can cause great therapeutic problems. High dosed corticosteroids are the treatment of choice. However, recalcitrant pyoderma gangraenosum or side effects from corticosteroid treatment may require therapeutic alternatives. Pyoderma gangraenosum responds excellently to treatment with cyclosporine A. Because of side effects and drug interactions, the patients must be selected and carefully and closely monitored. Six patients with pyoderma gangraenosum, unresponsive to various topical and systemic therapies, were treated with oral cyclosporine A at mean daily doses of approximately 3 mg/kg. Marked improvement of the skin lesions and complete healing occurred in all patients over a period of 3-6 months. Only one patient suffered a relapse after discontinuation of the treatment. No severe irreversible side effects occurred. The results show that low-dose cyclosporine A treatment can be considered a first line treatment of pyoderma gangraenosum. PMID- 7499131 TI - [Vorner keratosis palmoplantaris diffusa. Clinical, formal genetic and molecular biology studies of 22 families]. AB - In 1901, Vorner described a diffuse keratoderma of palms and soles with autosomal dominant inheritance. Histopathologically, this disease has the typical features of epidermolytic hyperkeratosis. Clinical examination does not allow differentiation between keratoderma of the Vorner type and the keratoderma described by Thost in 1880 and Unna in 1883. Reexamination of the family originally seen by Thost revealed histopathological signs of epidermolytic hyperkeratosis, confirming that keratoderma of the Vorner type is present in this family. The clinical features and variability of this palmoplantar keratoderma were demonstrated on the basis of an examination of 22 families (46 patients). In addition to diffuse hyperkeratosis of palms and soles with a sharp demarcation and erythematous margin, some less well-known features, such as knuckle pad-like keratoses on the finger joints and clubbing of the nails were observed. A genetic analysis of the pedigrees suggests that new mutations causing this disorder rarely occur. Point mutations in the keratin 9 gene, which has been mapped to chromosome 17q21, can be a cause of epidermolytic keratoderma of palms and soles. Five different keratin 9 gene mutations were identified. All these mutations are localized in the highly conserved coil 1A region of the rod domain, which is thought to be relevant for dimer formation in intermediate filaments. PMID- 7499130 TI - [Lymphocyte stimulation test, a possible alternative for verifying chloroacetophenone sensitization]. AB - We report on a case of pronounced sensitization to chloroacetophenone tear gas that developed after repeated occupational skin exposure in a 57-year-old police officer. Mainly in the presence of moisture and occlusion, cutaneous application of chloroacetophenone leads to severe irritant, and often also allergic, skin reactions. In patch testing the demonstration of allergic contact dermatitis in response to chloroacetophenone is hampered by the irritative potential of this substance even at low concentrations. This diagnostic bias can be overcome by the lymphocyte proliferation assay. PMID- 7499132 TI - [Removal of amateur and dirt tattooing with the Q-switched ruby laser]. AB - The Q-switched ruby laser with a wavelength of 694 nm and a pulse duration of 25 ns causes selective damage to skin pigment. This laser has produced good to excellent results in the treatment of amateur professional and traumatical tattoos, and also of benign pigmented lesions such as lentigines, ephelides, cafe au-lait spots and Becker nevi. We report on three patients with amateur and traumatic tattoos, which were removed by Q-switched ruby laser treatment without leaving scars. PMID- 7499133 TI - [Gianotti-Crosti syndrome in Epstein-Barr virus infection]. AB - Gianotti-Crosti syndrome, a relatively rare, distinctive eruption occurring after hepatitis B infection, is characterized by a lichenoid papular exanthema, usually localized on the face, limbs, and buttocks. Hepatitis B antigenaemia is associated with Gianotti-Crosti syndrome only in some cases. Recent reports indicate that a variety of infectious agents are associated with Gianotti-Crosti syndrome. This is a report of 2 1/2-year-old girl with Gianotti-Crosti syndrome and concurrent primary Epstein-Barr virus infection without evidence of hepatitis B infection. PMID- 7499134 TI - [Dermatomyositis-like skin changes with long-term hydroxyurea (Litalir) therapy]. AB - We report on dermatomyositis-like adverse cutaneous reactions following long-term maintenance therapy with hydroxyurea in two patients suffering from chronic myelogenous leukaemia (CML). In addition to non-specific side effects, such as xerosis, pruritus and hyperpigmentation, both patients presented with more specific skin changes, i.e. erythematous lesions, scaling, and partially atrophic areas distributed in a linear fashion on the dorsal aspects of the hands and fingers. In addition, teleangiectatic erythema of the face was present in both patients, and this was associated with oedema of the eyelids in one patient. Despite these dermatomyositis-like features there were no clinical signs of muscular involvement, and muscle-specific enzymes were within normal ranges. Skin biopsy specimens revealed an interface dermatitis characterized by a lichenoid cell infiltrate, vacuolar alteration of basal cells, necrotic keratinocytes within the spinous zone, focal hypergranulosis, ortho-hyperkeratosis and telangiectases in the upper part of the dermis. Analogous histopathological findings have been documented in lichen planus-like skin changes on the hands following hydroxyurea therapy. It seems doubtful whether there are actually any major differences between those skin changes described as dermatomyositis-like and those interpreted as lichen planus-like in patients receiving long-term hydroxyurea therapy. PMID- 7499135 TI - [Abdominal variant of benign symmetrical Launois-Bensaude lipomatosis]. AB - We report on a patient with an abdominal type of benign symmetrical Launois Bensaude lipomatosis. New aspects of the pathogenesis of abnormal distribution of the fatty tissue are also discussed. PMID- 7499136 TI - [Cutaneous calciphylaxis in chronic renal failure]. AB - A case of a 53-year-old chronically uraemic female patient who suddenly developed widespread cutaneous necrosis on both thighs is presented. On the basis of the typical histopathological features a diagnosis of calciphylaxis was made and supported by elevated serum parathyroid hormone levels. Calciphylaxis is a rare and serious disease, mostly seen in uraemic patients subjected to chronic haemodialysis. Typical features are calcifying panniculitis and thrombosis of small to medium-sized blood vessels followed by necrosis of the skin. The poor prognosis is due to systemic calcifications of visceral organs. When acute cutaneous necrosis of the skin is encountered in patients with chronic uraemia calciphylaxis should always be considered as a possible cause in the differential diagnosis. Determination of the serum parathyroid hormone level is recommended. PMID- 7499137 TI - [Histogenesis of nevus cell nevi. Implications of a case report]. AB - A case of multiple lesions of junctional melanocytic hyperplasia arising in a skin graft after excision of a congenital melanocytic naevus is reported. This observation is shown to support the concept of the histogenesis of acquired melanocytic naevi proposed by Cramer [2], i.e. that acquired melanocytic naevi are derived from melanocyte precursors located in nerve sheaths, whereas it is inconsistent with notions of the origin of naevogenesis in the epidermal melanocyte. PMID- 7499138 TI - [Increased incidence of dermatoses in nicotine and alcohol abuse]. PMID- 7499139 TI - [Gluten-free diet in Duhring dermatitis herpetiformis]. PMID- 7499141 TI - [Latex allergy]. PMID- 7499140 TI - [70 years Jena University Dermatologic Clinic--tradition and progress]. AB - The Department of Dermatology in Erfurter Street was founded in 1924. A short report is given about the early years of dermatology and the history of the clinic building in Jena. Its foundation is closely associated with Bodo Spiethoff, the first chairman of the Department of Dermatology at Jena University. PMID- 7499142 TI - Occurrence of radon in Riyadh groundwater supplies. AB - A study of 222Rn levels in the ground water supplies of the capital city of Saudi Arabia (Riyadh) was carried out. Such sources contribute up to 34% of the total city water demand. Samples were collected from 90 wells (75 deep and 15 shallow) out of 161 that supply water to six water treatment plants and from three locations within each plant. All samples have low radon levels with an average concentration of 2.99 +/- 0.29 and 3.44 +/- 0.35 Bq L-1 (61.8 +/- 7.8 and 92.9 +/ 9.5 pCi L-1) for the deep and shallow well waters, respectively. Due to water cooling, radon level reduction in the treatment plants was found to be in the range of 74-96%. The plants' product waters were found to have very low levels of radon with an overall average value of 0.49 +/- 0.14 Bq L-1 (13.2 +/- 3.8 pCi L 1). PMID- 7499143 TI - Estimated dose to a flood clean-up worker. AB - A worker involved in flood clean-up during the winter of 1994-1995 was inadvertently exposed to gamma rays from 226Ra and progeny. She worked near the radium, in the form of 10 radium needles, for 2.5 d, until it was identified and removed. Her external exposure was reconstructed and estimated to be 1.25-1.75 mSv (125-175 mrem). The activity of the radium needles (30 mCi each) was estimated by measuring the exposure rate at 1 m. PMID- 7499144 TI - Development of a system to evaluate and communicate radiation risk. AB - Review of research protocols involving positron emission tomography studies on healthy volunteers focused attention on the radiation exposure disclosure statements contained in the informed consent form. Of particular concern was the observation that breast doses from positron emission tomography studies are greater than breast doses from other research uses of radioisotopes, as well as routine nuclear medicine and radiographic procedures. Disclosure of individual organ doses is not normally provided on informed consent forms. A worksheet was developed to aid research investigators in the determination of effective dose equivalents and organ dose equivalents from all sources of radiation to which a volunteer is exposed. Three standardized risk statements are discussed. The final selection and use of these statements are determined by worksheet calculations of effective dose equivalents and organ dose equivalents. PMID- 7499145 TI - Consistency in radon dosimetry and nomenclature. PMID- 7499147 TI - Model testing using Chernobyl data: III. Atmospheric resuspension of radionuclides in Ukrainian regions impacted by Chernobyl fallout. AB - The "Resuspension" scenario is designed to test models for atmospheric resuspension of radionuclides from contaminated soils. Resuspension can be a secondary source of contamination after a release has stopped, as well as a source of contamination for people and areas not exposed to the original release. The test scenario describes three exposure situations: (1) locations within the highly contaminated 30-km zone at Chernobyl, where exposures to resuspended material are probably dominated by local processes; (2) an urban area (Kiev) outside the 30-km zone, where local processes include extensive vehicular traffic; and (3) a location 40 to 60 km west of the Chernobyl reactor, where upwind sources of contamination are important. Input data include characteristics of the 137Cs ground contamination around specific sites, climatological data for the sites, characteristics of the terrain and topography, and locations of the sampling sites. Predictions are requested for average air concentrations of 137Cs at specified locations due to resuspension of Chernobyl fallout and for specified resuspension factors and rates. Test data (field measurements) are available for all endpoints. PMID- 7499146 TI - Model testing using Chernobyl data: II. Assessment of the consequences of the radioactive contamination of the Chernobyl Nuclear Power Plant cooling pond. AB - The "Cooling Pond" scenario is designed to test models for radioactive contamination of aquatic ecosystems, based on data for contamination of different aquatic media and biota due to fallout of radionuclides into the cooling pond of the Chernobyl Nuclear Power Plant. Input data include characteristics of the cooling pond ecosystem (hydrological, hydrochemical, and hydrobiological conditions) and estimates of the amounts of 137Cs in the cooling pond. Predictions are requested in two stages: (1) calculations for 137Cs concentrations for comparison against actual measurements, including activities of 137Cs in the cooling pond water, in sediment layers, and in fish; and (2) calculations for which actual measurements are not available, including dose and risk estimates for aquatic biota and for humans following hypothetical consumption of contaminated biota. The latter calculations are intended to provide an opportunity for intercomparison among modelers of their results for a simulated assessment problem. PMID- 7499148 TI - Single point aerosol sampling: evaluation of mixing and probe performance in a nuclear stack. AB - Alternative reference methodologies have been developed for sampling of radionuclides from stacks and ducts, which differ from the methods previously required by the United States Environmental Protection Agency. These alternative reference methodologies have recently been approved by the U.S. EPA for use in lieu of the current standard techniques. The standard EPA methods are prescriptive in selection of sampling locations and in design of sampling probes whereas the alternative reference methodologies are performance driven. Tests were conducted in a stack at Los Alamos National Laboratory to demonstrate the efficacy of some aspects of the alternative reference methodologies. Coefficients of variation of velocity, tracer gas, and aerosol particle profiles were determined at three sampling locations. Results showed that numerical criteria placed upon the coefficients of variation by the alternative reference methodologies were met at sampling stations located 9 and 14 stack diameters from the flow entrance, but not at a location that was 1.5 diameters downstream from the inlet. Experiments were conducted to characterize the transmission of 10 microns aerodynamic diameter liquid aerosol particles through three types of sampling probes. The transmission ratio (ratio of aerosol concentration at the probe exit plane to the concentration in the free stream) was 107% for a 113 L min-1 (4-cfm) anisokinetic shrouded probe, but only 20% for an isokinetic probe that follows the existing EPA standard requirements. A specially designed isokinetic probe showed a transmission ratio of 63%. The shrouded probe performance would conform to the alternative reference methodologies criteria; however, the isokinetic probes would not. PMID- 7499149 TI - Dose equivalent measurements at a 2.7 GeV proton accelerator and comparison with the Moyer model. AB - The Moyer model, based on a semi-empirical method validated in the 7.4 to 350 GeV energy range, is generally used to calculate lateral shielding for high energy proton accelerators. Measurements made for the Saclay Synchrocyclotron, Saturne, have enabled the parameters corresponding to a 2.7 GeV model to be studied for different target thicknesses and angles of observation. These studies show how new data have been used to modify the equations of the model. PMID- 7499150 TI - A comparison of four aqueous-miscible liquid scintillation cocktails with an alpha/beta discriminating Wallac 1415 liquid scintillation counter. AB - This work was directed towards an assessment of alpha/beta separation, alpha detection efficiency, energy resolution, quench sensitivity, and background count rate for four commercially available liquid scintillation cocktails using a Wallac 1415 liquid scintillation counter. The four cocktails were Wallac Oy OptiPhase HiSafe 3 and HiSafe 2, and Packard Ultima Gold AB and Ultima Gold XR. Aqueous radioactive solutions were prepared in 0.1 N HCl and contained 241Am (5.49 MeV alpha) and 90Sr/90Y (0.46/2.28 MeV beta-). The radioactive solutions were loaded into the cocktails at a ratio of 1 mL of aqueous solution to 10 mL of cocktail in 20 mL high-density polyethylene (HDPE) liquid scintillation vials. The effect of three levels of chemical quench on the various properties of interest was assessed for each of the LS cocktails. Alpha/beta discrimination was quantified by the fraction of beta interactions that "spillover" into the alpha window and vice versa, at the working discriminator setting. The working discriminator setting was defined as the point where the spillover of beta interactions into the alpha window and alpha interactions into the beta window were equal. For the low-quench samples, Ultima Gold AB had the lowest spillover (0.25%) and Ultima Gold XR had the highest spillover (0.8%). For the high-quench samples, the spillover ranged from 4% (HiSafe 3) to 10.5% (Ultima Gold XR). The detection efficiency for 241Am was nearly 100% and decreased with increased quench because of loss from spillover. HiSafe 2 gave the best energy resolution, 500 keV (FWHM), for the low-quench sample. The lowest background count rate was achieved with Ultima Gold AB, 0.056 cpm in the 241Am window, for a low-quench blank. PMID- 7499151 TI - Development of an atmospheric 222Rn concentration model using a hydrodynamic meteorological model: I. One-dimensional practical model. AB - A one-dimensional numerical model for 222Rn transport in an atmospheric boundary layer was developed. The model consists of two parts: a prognostic hydrodynamic model including a turbulence closure model and an atmospheric diffusion model for 222Rn. The first part predicts meteorological conditions to provide the second part with vertical turbulence conditions which affects the vertical motion of 222Rn near the Earth's surface. Calculations with the model are compared with 222Rn concentrations measured during a variety of meteorological conditions, from clear days with high radiation and low winds to cloudy days of low radiation with high winds. The model's results represent well the typical diurnal variations of the 222Rn concentrations. PMID- 7499153 TI - Development of an atmospheric 222Rn concentration model using a hydrodynamic meteorological model: II. Three-dimensional research-purpose model. AB - This paper describes a new type of three-dimensional numerical model for 222Rn transport in an atmospheric boundary layer. The model is a combination of a prognostic hydrodynamic meteorological model including a turbulence closure model and an atmospheric diffusion model for 222Rn. The first part provides the second part with the meteorological conditions needed for calculations of the 222Rn transport and diffusion. The model is capable of giving consideration to horizontal source distribution, complex terrain, and non-uniform and non-steady atmosphere. The model's results were compared with 222Rn field measurements in a mountain-valley area and represented qualitatively a typical diurnal variation of the 222Rn concentration in nocturnal drainage flows. The model was also applied to the transport of 222Rn in a seacoast area. These results indicated that the model could be effective as a research tool for numerical analysis of 222Rn behavior under various atmospheric conditions. PMID- 7499152 TI - Opportunities for the testing of environmental transport models using data obtained following the Chernobyl accident. AB - The aftermath of the Chernobyl accident has provided a unique opportunity to collect data sets specifically for the purpose of model testing, and with these data to create scenarios against which environmental transport models may be tested in a format constituting a blind test. This article serves as an introduction to three test scenarios designed for testing models at the process level: (1) surface water contamination with radionuclides initially deposited onto soils; (2) contamination of different aquatic media and biota due to fallout of radionuclides into a body of water; and (3) atmospheric resuspension of radionuclides from contaminated land surfaces. These scenarios are the first such tests to use data sets collected in the former Soviet Union. Interested modelers are invited to participate in the test exercises by making calculations for any of these test scenarios. Information on participation is included. PMID- 7499154 TI - Survey of 222Rn concentrations in dwellings and soils in the Dutch Belgian border region. AB - 222Rn levels in dwellings and soil gas have been investigated in the Eijsden-Vise region, located at the Dutch-Belgian border, in order to analyze the relationship between domestic radon levels and soil gas radon levels, in view of the fact that the local geology varies substantially from uranium-rich to uranium-poor rocks and soils. During February 1992, charcoal detectors were exposed for 24 h in 116 dwellings in the township of Vise, a radon prone area in Belgium. As expected, the results show a large regional variation, with an averaged indoor air radon level of 116 Bq m-3. In the nearby township of Eijsden, the Netherlands, an area with a lower radiation level, similar measurements by means of charcoal detectors in 42 dwellings during March 1993, resulted in an averaged indoor 222Rn concentration of 46 Bq m-3. Furthermore, in the same region time-integrated radon measurements were performed in 15 dwellings on different floors. These results indicate that a gradient in indoor air radon concentration exists from basement to upper floor level. In addition, a significant positive correlation has been found between radon gas levels of soils surrounding 26 houses and indoor air radon levels. The variation in indoor air radon levels in the region under study is mainly due to the variation of the radon levels in the soils. A soil gas radon survey is suitable for prescreening of the radon potential of an area taking into account the spatial heterogeneity of the local geology. PMID- 7499155 TI - Natural radionuclide and plutonium content in Black Sea bottom sediments. AB - The content of uranium, thorium, radium, lead, polonium, and plutonium in bottom sediments and algae from two locations at the Bulgarian Black Sea coast have been determined. Some parent:progeny ratios for evaluation of the geochemical behavior of the nuclides have been estimated as well. The extractable and total uranium and thorium are determined by two separate radiochemical procedures to differentiate the more soluble chemical forms of the elements and to estimate the potential hazard for the biosphere and for humans. No distinct seasonal variation as well as no significant change in total and extractable uranium (also for 226Ra) content is observed. The same is valid for extractable thorium while the total thorium content in the first two seasons is slightly higher. Our data show that 210Po content is accumulated more in the sediments than 210Pb, and the evaluated disequilibria suggest that the two radionuclides belong to more recent sediment layers deposited in the slime samples compared to the silt ones for the different seasons. The obtained values for plutonium are in the lower limits of the data cited in literature, which is quite clear as there are no plutonium discharge facilities at the Bulgarian Black Sea coast. The obtained values for the activity ratio 238Pu:239 + 240Pu are higher for Bjala sediments compared to those of Kaliakra. The ratio values are out of the variation range for the global contamination with weapon tests fallout plutonium which is probably due to Chernobyl accident contribution. The dependence of natural radionuclide content on the sediment type as well as the variation of nuclide accumulation for two types of algae in two sampling locations for five consecutive seasons is evaluated. No serious contamination with natural radionuclides in the algae is observed. PMID- 7499156 TI - Model testing using Chernobyl data: I. Wash-off of 90Sr and 137Cs from two experimental plots established in the vicinity of the Chernobyl reactor. AB - The "Wash-off" scenario is designed to test models concerned with the movement of trace contaminants from terrestrial sources to bodies of water, specifically the contamination of surface water by wash-off of radionuclides initially deposited onto soils. Particular emphasis is placed on chemical speciation and on the geochemical and geophysical processes affecting transfer of contaminants from soil to water. The scenario gives descriptions of two experimental plots near the Chernobyl power plant, one using heavy rain and one using snow melt, together with characteristics of the initial aerial deposition of the radionuclides and data on topography, soil type and characteristics, and time-varying precipitation. Predictions are requested for (1) the vertical distribution of concentrations of exchangeable and nonexchangeable forms of 137Cs and 90Sr in the soil of the experimental plots, (2) concentrations of 137Cs and 90Sr in runoff water from the experimental plots, and (3) total amounts of 137Cs and 90Sr removed by runoff from the experimental plots. Test data (field measurements) are available for all endpoints. PMID- 7499157 TI - Sex-specific tissue weighting factors for effective dose equivalent calculations. AB - The effective dose equivalent was defined in the International Commission on Radiological Protection Publication 26 in 1977 and later adopted by the U.S. Nuclear Regulatory Commission. To calculate organ doses and effective dose equivalent for external exposures using Monte Carlo simulations, sex-specific anthropomorphic phantoms and sex-specific weighting factors are always employed. This paper presents detailed mathematical derivation of a set of sex-specific tissue weighting factors and the conditions which the weighting factors must satisfy. Results of effective dose equivalent calculations using female and male phantoms exposed to monoenergetic photon beams of 0.08, 0.3, and 1.0 MeV are provided and compared with results published by other authors using different sex specific weighting factors and phantoms. The results indicate that females always receive higher effective dose equivalent than males for the photon energies and geometries considered and that some published data may be wrong due to mistakes in deriving the sex-specific weighting factors. PMID- 7499159 TI - Radon mitigation at Birch Cliff Public School. AB - In 1991, Canadian Institute for Radiation Safety (CAIRS) conducted a radon screening program in all Metropolitan Toronto public schools. Birch Cliff Public School had a radon progeny level higher than the action level of 4.16 x 10(-7) Jm 3 (20 mWL). Follow-up radon testing was carried out at the school. Locations on the ground floor and in the basement were tested. All locations on the ground floor had radon progeny levels below the action level. Six locations in the basement had readings above the action level. All cracks and openings in the basement were sealed and a new heating/ventilating (HV) system for the basement was designed and installed. Then, the basement was tested again. Radon progeny levels are now well below the action level with an average of 7.43 x 10(-8) Jm-3 (3.57 mWL). This is about one fifth of the average radon progeny level found in the first stage of follow-up testing. PMID- 7499158 TI - Effective doses due to external irradiation from the Chernobyl accident for different population groups of Ukraine. AB - A model for the external exposure of the Ukrainian population after the Chernobyl accident was developed. It is based on extensive measurements of external gamma exposure rates (EGER) in air and of external effective doses of members of five population groups. Questionnaires were used to determine the occupancy times of members of the population groups at three types of locations; inside houses, outdoors, and outside of the home settlement. Behavior factors are defined as the ratio of individual external doses to a reference dose for a phantom standing permanently over an open field with the same average 137Cs activity per unit area as in the settlement. The behavior factors were derived for five population groups (children younger than seven years, the age group from eight to seventeen years, employees, agricultural workers, and pensioners) by two methods: first from direct measurements of individual doses by thermoluminescent dosimetry and an experimental determination of the average 137Cs activity per unit area in the settlement of interest; and second from external gamma-exposure rates in air at various types of locations and from the questionnaire data. The methods were found to be consistent and the results were used to calculate external exposures of the five population groups in the years 1987 through 1991. PMID- 7499160 TI - [100 years of radiotherapy in otorhinolaryngology]. PMID- 7499161 TI - [1995 ENT Congress in Karlsruhe. The cranial base]. PMID- 7499162 TI - [Modern neuroradiologic diagnosis and interventional strategies for the ENT physician]. AB - The value of imaging and interventional neuroradiological methods in ENT is surveyed. The role of CT and MRI is discussed in the diagnosis of various diseases. CT remains the method of choice in inflammatory paranasal sinus disease and in craniofacial trauma, whereas MRI has become the preferred imaging technique in patients with sensorineural hearing loss. CT and MRI are complementary in the assessment of skull base tumors. Preoperative embolization facilitates surgery of highly vascular tumors. Endovascular procedures play a prominent role in the control of hemorrhages of the head and neck region and are the method of choice in the treatment of arteriovenous fistulas. PMID- 7499163 TI - [Modern radiologic diagnosis for the ENT physician]. AB - Radiologic diagnosis has acquired an increased importance in head and neck diseases since the introduction of sectional imaging techniques. In addition to clinical and endoscopic examinations, CT has become the standard modality for TNM staging of malignant diseases. CT is also a basic tool in diagnosing benign diseases localized in or infiltrating into the deep head and neck regions. Ultrasound is widely available and is now well established in the diagnosis of superficial pathologies. Magnetic resonance is the newest imaging modality and is being increasingly used in clinical practice. By avoiding ionizing radiation MRI provides an improved method for contrast studies and enables high-quality multiplanar imaging. The applications of this method are still expanding with ongoing refinements of MR imaging techniques. Conventional radiological methods remain limited to a few indications. Radiation dose and cost have now required replacement of traditional step-by-step diagnostic methods by the one method that can meet most requirements in a single examination. PMID- 7499164 TI - [3-dimensional imaging of temporal bone structures using spiral CT. Initial results in normal temporal bone anatomy]. AB - 3D reconstruction of the temporal bone using spiral CT techniques was performed in 51 patients with various otological diseases during routine clinical work evaluation. The 3D display was optimized by a reduced study time and improved detail accuracy by special algorithms. We were able to demonstrate comprehensively in a 3D mode the normal anatomy of the inner ear and adjacent middle ear structures, such as the modiolus of the cochlea, the semicircular canals, the cochlear and vestibular aqueduct and the ossicles. We suggest routine 3D delineation of the substructures of the temporal bone prior to otologic surgery to provide the surgeon with a 3D view of individual anatomy and specific otosurgical sites. PMID- 7499165 TI - [The "dangerous loop" of the internal carotid artery. An anatomic study]. AB - Variations of the clinical course of the internal carotid artery in the parapharyngeal space were studied in order to define possible causes of acute hemorrhage during pharyngeal surgery. Four cases of sigmoid tortuosity involving the internal carotid artery were found in a total of 81 dissected carotid sheathes and 8 corrosion casts. In 2 cases the internal carotid artery was located in direct contact with the tonsillar fossa to form a clinically unsafe carotid tortuosity that was considered at risk during tonsil surgery. PMID- 7499166 TI - [Results and surgical experiences in web transsection of hypopharyngeal (Zenker) diverticulum with the CO2 laser]. AB - Since 1984 at the Department of Otorhinolaryngology, Head and Neck Surgery, University of Kiel, 34 patients with hypopharyngeal diverticular have been operated microendoscopically using the CO2 laser. Results and complications of this technique were compared to a group of patients in whom standard (non-laser) diverticulotomy was carried out from 1976 to 1993. Ninety per cent of the patients treated endoscopically with laser surgery have not had complaints and the rate of postoperative complications has been low. Three patients developed pharyngeal stenoses while one case incurred a mediastinitis. Intraoperative bleeding was not observed. In comparison to the microendoscopic procedure serious complications were more frequently observed in patients treated with the standard diverticulotomy. In eight of these cases paralysis of the recurrent laryngeal nerve occurred. A mediastinotomy was performed in another three patients because of mediastinitis. In two cases stenosis occurred after surgery and one patient developed a fistula. Endoscopic diverticulotomy utilizing a CO2 laser was found to be an excellent treatment for Zenker's diverticulum because of the good functional results achieved and low complication rate. The procedure was easy to perform because of the short duration and could be repeated frequently without significant additional risks. PMID- 7499167 TI - [Effect of variations in middle ear pressure on ruptures of the round window membrane of the inner ear of the guinea pig (Cavia porcellus)]. AB - The electrical activity of the inner ear before and after rupture of the round window membrane was monitored in guinea pigs under different pressure conditions with the aid of electrocochleography. Following studies were conducted weekly over a period of 4 weeks. Findings showed that overpressure below the pressure needed to open the Eustachian tube caused strong temporary functional disturbance of the cochlea, especially at high frequencies. Irreversible changes that were pressure-dependent were observed and occurred mainly at high frequencies. Application of low pressure to the round window membrane caused a functional loss comparable to changes after overpressure. Animals with a predamaged cochlea reacted to overpressure that was below opening pressure of the tube and to corresponding low pressure with a longer lasting functional loss than did animals with an undamaged cochlea. PMID- 7499168 TI - [Necrotizing fasciitis of soft tissues of the face and neck]. AB - Three cases of cervical necrotizing fasciitis are presented. One case was odontogenic in origin and two were due to pharyngeal infectious. Bacteria cultured represented multiple bacterial species. Airway control was necessary early, as was wide surgical exploration of the fascial spaces of the neck with re exploration as necessary. Intensive medical support was crucial to prevent or treat complications. Cervical necrotizing fasciitis has an overall mortality rate of 30 per cent at the University of Bern. PMID- 7499169 TI - [Primary pleomorphic adenoma of the principal paranasal sinus]. AB - The pleomorphic adenoma is the most frequent benign tumor occurring in the salivary glands. These tumors are very uncommon in the nasal fossae and paranasal sinuses. We report a case of pleomorphic adenoma in the nasal fossa and review the currently available literature. Different tumor localizations and therapeutic concepts are discussed. PMID- 7499170 TI - [Diagnosis and therapy of salivary gland tumors. I]. PMID- 7499171 TI - HLA-DR residues accessible under the peptide-binding groove contribute to polymorphic antibody epitopes. AB - Many residues involved in polymorphic antibody-binding epitopes on class II molecules are located on the alpha-helix of DR beta chains. Although they have received less attention, residues in the peptide-binding groove and second domain of the DR beta chain may also be critical for polymorphic anti-DR antibody epitopes. In this study, we used transfectants expressing site-directed mutations at positions in the HLA-DR beta 1 and beta 2 domains and flow cytometry to define the epitopes of several polymorphic anti-DR antibodies. Both DR(beta 1*0403) residues 14 and 25 were shown to be involved in the epitopes of mAbs DA6. 164, HU 20, Q5/6, and 50D6, and DR(beta 1*0701) residue 14 was shown to be critical for the epitopes of two DR7-specific mAbs, SFR 16-DR7M and TAL13.1. Unlike most other residues shown to be important in antibody-binding epitopes, residue 14 is located in the floor of the peptide-binding groove and residue 25 is in an outer loop, each with their side chains pointing down, such that antibodies may directly contact these residues from below the binding groove. Two residues in the beta 2 domain, beta 180 and beta 181, were also shown to be involved in the epitopes of three polymorphic anti-DR mAbs, NFLD.D1, NFLD.M1, and LY9. Although these two residues are close to the transmembrane domain in the linear sequence, their solvent accessibility in the DR1 structures is quite impressive. Our data provide new evidence that residues accessible under the peptide-binding groove contribute to polymorphic antibody-binding epitopes. PMID- 7499172 TI - Ultrastructural allelic variation in HLA-DQB1 promoter elements. AB - Sequence variation among HLA class II promoter elements may contribute to functional differences in transcriptional regulation of different class II alleles. In addition to influencing the binding sites for nuclear transcription factors, promoter polymorphism may also alter intrinsic structural properties of the DNA strands, such as conformation and curvature, which influence the formation of stable transcription complexes. We used SSCP analysis of PCR amplified promoter regions from the DQB1 locus to evaluate conformational polymorphism within DQ alleles. Distinct electrophoretic migration patterns of the SSCP products were detected for six DQB1 alleles; analysis of the DQB1*0302 promoter, known to be associated with type 1 diabetes, showed no SSCP differences between IDDM patients and normal controls. Using computer modeling based on a "nearest-neighbor" energy of predicted curvature theory, we examined the effect of allelic promoter region sequence polymorphism on the predicted curvature of double-stranded DNA, and found distinct allelic differences in predicted DNA curvature, both in transcriptional consensus binding sites and in regions located between binding sites. These data are consistent with a model in which intrinsic sequence variation in the promoter region results in ultrastructural differences which may influence DNA bending and interactions with multimeric DNA-protein transcription complexes. PMID- 7499173 TI - HLA allele and haplotype frequencies in Algerians. Relatedness to Spaniards and Basques. AB - The powerful genetic polymorphism of the HLA system has been used to identify individuals and populations. Ethnic groups may be characterized by specific HLA allele frequencies and particular extended HLA haplotypes; also, genetic relationships among these groups may be deduced. In the present study, serology and DNA typing were used to detect HLA-A, -B, -C, -DR, and -DQ alleles in each individual and to calculate characteristic haplotypes in Algerians. These results were compared to those previously obtained in other populations, particularly northern Mediterraneans; genetic distances and their respective dendrograms place Basques and Spaniards closer to Algerians than to other Europeans. Also, characteristic Basque and/or Spanish haplotypes are found in Algerians; i.e., A30 B18-Cw5-DR3-DQ2 and A1-B57-Cw7-DR7-DQ2. This supports the evidence that the Algerian population, mainly its paleo-North African component (Berbers), has a common descent with Basques and Spaniards, probably reflecting a preneolithic relationship between Iberians and paleo-North Africans. PMID- 7499175 TI - Three highly polymorphic microsatellites at the human myelin oligodendrocyte glycoprotein locus, 100 kb telomeric to HLA-F. Characterization and relation to HLA haplotypes. AB - The MOG locus, located on chromosomal bands 6p21.3-p22 and mapped about 100 kb telomeric to HLA-F, was isolated from cosmid ICRFc109A2434 and shown to contain three microsatellites. These CA-repeat polymorphic markers were characterized in a sample of 173 healthy unrelated individuals and 84 DNAs from the HLA Workshop reference panel, by a method combining fluorescence labeling of PCR products and use of an automated DNA sequencer. For the three markers, frequencies of heterozygotes are well predicted from allele frequencies by the Hardy-Weinberg rule, which suggests that problems of allele nonamplification are unlikely. Typing of cell lines homozygous in the HLA region allowed unambiguous definition of 81 HLA-MOG haplotypes and showed that several HLA ancestral haplotypes extended to the MOG region. The high degree of polymorphism (59%, 51%, and 81% at the three loci, respectively, and 87% at the haplotype level) makes these new markers informative for association or linkage studies with diseases such as hemochromatosis or multiple sclerosis, and for studies aimed at precisely delineating the site of crossover in chromosomes in which recombination occurred in the distal part of the HLA class I region. PMID- 7499174 TI - Characterization of human T-cell responses to Yersinia enterocolitica superantigen. AB - We reported that antigenic preparations from Yersinia enterocolitica stimulate murine T cells in a manner consistent with that of superantigens. As a consequence we examined whether Y. enterocolitica antigenic preparations stimulate human T-cell cultures. Human T cells, enriched from peripheral blood lymphocytes, were stimulated to proliferate in the presence of Y. enterocolitica cytoplasmic and membrane preparations. This activity has also been shown to be sensitive to protease treatment, indicating the presence of a protein, and when separated by ion-exchange chromatography a single peak of activity is resolved. Furthermore, this proliferation was inhibited, in a dose-dependent manner, by the presence of antibodies directed against MHC class II antigens, indicating a requirement for these molecules. When these cells were stained with a panel of V beta-specific antibodies to determine if there was an enrichment of a particular V beta-bearing T-cell subset after stimulation, results indicate a significant enrichment of T cells bearing V beta 3, V beta 12, V beta 14, and V beta 17 over controls. Taken together, these data are consistent with a Y. enterocolitica product acting as a superantigen for human T cells. PMID- 7499176 TI - CD3 delta and epsilon gene expression in CD3-CD16+ natural killer cell clones derived from thymic precursors. AB - To better understand the maturational stages during T-cell development, we studied the expression of CD3 delta and CD3 epsilon genes, as well as the presence of TCR gene rearrangements, within CD3-CD16+ NK clones derived from thymic precursors in vitro. Northern blot analysis revealed that CD3-CD16+ clones derived from CD7+CD3-CD4-CD8- (TN) thymocytes expressed transcripts for the CD3 epsilon gene; however, no transcripts for the CD3 delta gene were detected. Importantly, both the CD3 epsilon and CD3 delta genes were expressed in TN thymocytes examined prior to cloning. A CD7+CD8+CD3-CD4- thymocyte population that makes up only 0.4% of the total thymocyte pool was also isolated from human thymus. We determined the maturation potential of this CD7+CD8+CD3-CD4- population by limiting dilution cloning and found that 67% of the clones generated in vitro had a CD3-CD16+CD8+ phenotype. In contrast to the NK clones derived from TN precursors, most CD3-CD16+ clones derived from CD7+CD8+CD3-CD4- thymocytes expressed transcripts for both CD3 epsilon and CD3 delta genes. Southern blot analysis of the NK clones derived from either thymic precursor population revealed no rearrangement of the TCR beta or gamma genes. These results demonstrate that the TN progenitor population and their CD3-CD16+ progeny differ in their expression of the CD3 delta transcript and during in vitro culture there is loss of CD3 delta expression and acquisition of surface CD16 within these NK clones. Furthermore, the CD3-CD16+ clones derived from TN versus CD7+CD8+CD3-CD4- thymocytes differed in their expression of the CD3 delta gene. The signaling events regulating the expression of the CD3 invariant chain genes within immature lymphoid progenitor cells may be important in determining their eventual maturation into T-cell and NK-cell lineages in vivo. PMID- 7499177 TI - A new HLA-C allele, Cw*1403, associated with HLA-B44 in Japanese. AB - An allele encoding an HLA-C antigen, tentatively called CX44, associated with HLA B44 was identified as a new member of the Cw14 group, Cw*1403. The nucleotide sequence of Cw*1403 was closest to that of Cw*1401: five bases were different between the two alleles, in which three bases in Cw*1403 (two in exon 3 and one in exon 4) were the same as those of most HLA-C alleles. Two substitutions from guanine to adenine were found in the new allele, both of which are in exon 2, one at position 134 (61 of exon 2) and the other at position 201 (128 of exon 2). The former nucleotide substitution leads to the substitution of amino acid residue 21 from Arg to His, and the other substitution was synonymous. The former substitution was shared with Cw2, 3, 5, 13, and 15 alleles, and the latter was shared with Cw2, 4, 5, 8, 12, 13, 15 and 16 alleles. The other seven unrelated Japanese samples with CX44 were analyzed by a PCR-SSO method. It was confirmed that all the seven samples have the same substitutions as the sequenced allele, and the allele demonstrates a strong association with A33, B44, DR13, and DQ1, which are known to form a common haplotype in Japanese and Koreans. PMID- 7499179 TI - A novel HLA-DR13 allele (DRB1*1314) identified by single-strand conformation polymorphism analysis and confirmed by direct sequencing. AB - A novel variant of the HLA-DR13 group is described. The new allele was found in a DR10, 13 heterozygous patient of Turkish origin, two HLA genotypically identical children of the patient typed as DR11,13, and one child typed as DR13,13. DR13 subtyping of the patient was initially performed by SSCP analysis of PCR amplified DNA, using the 11th OHWS primers DRBAMP-3 and DRBAMP-B. Due to an unusual SSCP banding pattern, the PCR product was subjected to solid-phase sequencing. The sequence of the new allele, DRB1*1314, is different from that of DRB1*1307 by a single nucleotide substitution in codon 47, with T replacing consensus A. This results in a single amino acid change of tryptophan to phenylalanine in the first domain of the DR beta chain. PMID- 7499178 TI - The distribution of DR4 haplotypes in Sardinia suggests a primary association of type I diabetes with DRB1 and DQB1 loci. AB - The contribution of genetic variation at HLA class II loci to the susceptibility to and protection from IDDM was investigated by analyzing the distribution of HLA DRB1*04 haplotypes in 630 Sardinian newborns and 155 Sardinian IDDM patients. The different RRs and ARs of the various DR4-DQB1*0302 haplotypes, significantly ranging from the strongly associated DRB1*0405, DQB1*0302 to the protective DRB1*0403, DQB1*0302 haplotypes, provides clearcut evidence that the DRB1 locus is crucial in conferring IDDM predisposition or protection. Also, the DQB1 locus influences IDDM predisposition or protection by restricting the disease-positive association to DRB1*0405 haplotypes carrying the susceptibility DQB1*0302 or DQB1*0201 alleles but not the protective DQB1*0301 allele. Haplotype analysis not only suggests that the DRB1 and DQB1 loci influence IDDM risk in the same way, but also that the HLA-linked protection is "dominant" compared with "susceptibility." These results, obtained from a population with one of the highest IDDM incidences in the world, define more clearly the contribution of the various HLA loci to IDDM protection or susceptibility and allow a more precise calculation of AR. PMID- 7499181 TI - The safety of Bacillus species as insect vector control agents. PMID- 7499180 TI - Nomenclature for factors of the HLA system, update June 1995. PMID- 7499182 TI - Everybody's doing it: a P.E.T. peeve or a public service? PMID- 7499183 TI - Loss prevention response. PMID- 7499185 TI - Progress report: Arkansas Foundation for Medical Care as a quality foundation. PMID- 7499184 TI - Acute pancreatitis as the first manifestation of hyperparathyroidism in an otherwise asymptomatic patient. AB - Acute pancreatitis as a manifestation of hyperparathyroidism (HPT) has been reported in the literature but the concept of causal relationship has been disputed. We report a case of acute pancreatitis where the presence of hypercalcemia led to the diagnosis of primary HPT. No other current risk factors for pancreatitis were identified and no symptoms of HPT were present prior to this episode. We review the literature regarding the role of HPT in the pathogenesis of acute pancreatitis. PMID- 7499186 TI - Cutting red tape on clinical labs: why Congress should deregulate doctors. PMID- 7499187 TI - Pericardial effusion. PMID- 7499188 TI - Prenatal WIC participation related to Medicaid cost savings and infant health outcomes. PMID- 7499189 TI - Radiological case of the month. Pulmonary alveolar proteinosis. PMID- 7499190 TI - Preferential involvement of MEK1 in the tumor necrosis factor-alpha-induced activation of p42mapk/erk2 in mouse macrophages. AB - The pleiotropic cytokine tumor necrosis factor-alpha (TNF alpha) controls the expression of multiple gene products in macrophages and plays an important role in host defense. TNF alpha is recognized by the receptors, CD120a (p55) and CD120b (p75). Ligation of CD120a (p55) by TNF alpha or by anti-receptor agonistic antibodies initiates signal transduction leading to the activation of mitogen activated protein kinases (MAPKs) (p42mapk/erk2 and p44mapk/erk1). Phosphorylation and activation of MAPK are mediated by MAPK kinase (MEK), a family of Thr/Tyr kinases. In this study, we investigated the preferential involvement of the MEK isoforms MEK1 and MEK2 in the activation of p42mapk/erk2 in mouse macrophages stimulated with TNF alpha. Exposure of macrophages to TNF alpha stimulated a time-dependent increase in the activity of MEK1 as measured by an in vitro kinase assay using kinase-inactive p42mapk/erk2 (rMAPKkd) as substrate in the presence of gamma-[32P]ATP. Maximal activation of MEK1 was detected at 10 min poststimulation and coincided with maximal transphosphorylation of Tyr and Thr residues of rMAPKkd. By contrast, there was no evidence of MEK2 activation in macrophages in response to TNF alpha. These data suggest that MEK1 is the preferred substrate for MEK kinase, the upstream kinase implicated in activation of the MAPK pathway in macrophages by TNF alpha. PMID- 7499191 TI - Thrombin induces activation of p38 MAP kinase in human platelets. AB - In human platelets a proline-directed kinase distinct from the ERK MAP kinases is stimulated by both thrombin and the thrombin receptor agonist peptide SFLLRN and may be involved in the activation of Ca(2+)-dependent cytosolic phospholipase A2 (Kramer, R. M., Roberts, E. F., Hyslop, P. A., Utterback, B. G., Hui, K. Y., and Jakubowski, J.A. (1995) J. Biol. Chem. 270, 14816-14823). Here we show that this kinase is identical with or closely related to p38 (the mammalian homolog of HOG1 from yeast), a recently discovered protein kinase typically activated by inflammatory cytokines and environmental stress. Further, we demonstrate that activation of this kinase by thrombin is transient (with maximal stimulation at 1 min), is accompanied by tyrosine phosphorylation, and precedes the activation of the ERK kinases. This is the first report to show that p38 kinase is activated by thrombin and to suggest a role for this MAP kinase in the thrombin-mediated signaling events during platelet activation. PMID- 7499192 TI - Disruption of reconstituted nucleosomes. The effect of particle concentration, MgCl2 and KCl concentration, the histone tails, and temperature. AB - We find that reconstituted nucleosome cores containing specific DNA sequences dissociate on dilution. This disruption of histone-DNA contacts leading to the release of free DNA is facilitated by the presence of the core histone tails, MgCl2 (5 mM), KCl (60 mM), and temperatures above 0 degree C. Under reaction conditions that are commonly used to assess trans-acting factor access to nucleosomal DNA, histone-DNA contacts are on the threshold of instability. We demonstrate how dilution of reconstituted nucleosomes containing a TATA box can facilitate TBP access to DNA. PMID- 7499193 TI - Oligomerization of endothelial nitric oxide synthase. Evidence for a dominant negative effect of truncation mutants. AB - Nitric oxide produced by the endothelial isoform of nitric oxide synthase (ecNOS) is a key determinant of vascular tone. In contrast to other nitric oxide synthase (NOS) isoforms, which have been characterized as soluble homodimeric enzymes, ecNOS is predominantly membrane-associated, a feature that has hindered direct biochemical analyses of its oligomeric structure. We investigated ecNOS oligomerization using co-immunoprecipitation experiments in transiently transfected COS-7 cells. When COS-7 cells co-transfected with constructs encoding wild-type ecNOS and an epitope-tagged myristoylation-deficient mutant were biosynthetically labeled with [3H]myristate, the antibody to the epitope tag specifically immunoprecipitated 3H-labeled ecNOS, reflecting enzyme oligomerization. In COS-7 cells transfected with cDNAs encoding epitope-tagged truncation mutants and untagged full-length ec-NOS, the wild-type enzyme could be immunoprecipitated by the antibody to the epitope tag. Co-immunoprecipitation of ecNOS with truncation mutants documented that both N- and C-terminal domains are involved in ecNOS oligomerization. When these truncation mutants are co-expressed with wild-type ecNOS, they exert a marked dominant negative effect on enzyme activity. Since NOS oligomerization itself may be subject to dynamic modulation, the regulation of ecNOS assembly may have implications for NO signaling in the vascular wall. PMID- 7499194 TI - PTB domains of IRS-1 and Shc have distinct but overlapping binding specificities. AB - PTB domains are non-Src homology 2 (SH2) phosphotyrosine binding domains originally described in the receptor tyrosine kinase substrate, Shc. By serial truncation, we show that a 174-residue region of Shc p52 (33-206) has full PTB activity. We also show that a 173-residue region of insulin receptor substrate-1 (IRS-1; residues 144-316) has related PTB activity. In vitro both domains bind directly to activated insulin receptors. Binding is abrogated by substitution of Tyr-960 and selectively inhibited by phosphopeptides containing NPXY sequences. Phosphopeptide assays developed to compare PTB domain specificities show that the Shc PTB domain binds with highest affinity to psi XN beta 1 beta 2 pY motifs derived from middle T (mT), TrkA, ErbB4, or epidermal growth factor receptors (psi = hydrophobic, beta = beta-turn forming); the IRS-1 PTB domain does not bind with this motif. In contrast, both the Shc and IRS-1 PTB domains bind psi psi psi XXN beta 1 beta 2pY sequences derived from insulin and interleukin 4 receptors, although specificities vary in detail. Shc and IRS-1 are phosphorylated by distinct but overlapping sets of receptor-linked tyrosine kinases. These differences may be accounted for by the inherent specificities of their respective PTB domains. PMID- 7499196 TI - Roles of vaccinia virus ribonucleotide reductase and glutaredoxin in DNA precursor biosynthesis. AB - To examine the possible role of the vaccinia virus glutaredoxin as a cofactor for viral ribonucleotide reductase, viral growth, DNA synthesis, and dNTP pools were measured in infections of B-SC-40 monkey kidney cells with wild type vaccinia virus and with mutants of vaccinia that lacked a functional reductase or glutaredoxin. In infections of untreated host cells, the lack of viral ribonucleotide reductase or glutaredoxin had only small effects upon virus growth. When host cells were pretreated with alpha-amanitin, which blocks host RNA polymerase II but not viral transcription, viral DNA synthesis was markedly reduced in infections with either of the mutants when compared with wild type infections. Relative to dNTP levels in wild type infections, pools of dCTP, but not of the other dNTPs, were significantly reduced in infections of amanitin treated cells with either mutant. The parallel depletion of dCTP in the two mutant suggests that the role of glutaredoxin may be to function as a cofactor for viral ribonucleotide reductase. The data suggest that both viral proteins become essential for DNA replication only when levels of the corresponding host cell proteins are depleted. PMID- 7499195 TI - Molecular cloning and functional expression of a novel amiloride-sensitive Na+ channel. AB - We have isolated a cDNA for a novel human amiloride-sensitive Na+ channel isoform (called delta) which is expressed mainly in brain, pancreas, testis, and ovary. When expressed in Xenopus oocytes, it generates an amiloride-sensitive Na+ channel with biophysical and pharmacological properties distinct from those of the epithelial Na+ channel, a multimeric assembly of alpha, beta, and gamma subunits. The Na+ current produced by the new delta isoform is increased by two orders of magnitude after coexpression of the beta and gamma subunit of the epithelial Na+ channel showing that delta can associate with other subunits and is part of a novel multisubunit ion channel. PMID- 7499197 TI - Ionizable P1 residues in serine proteinase inhibitors undergo large pK shifts on complex formation. AB - The burial of charged residues in proteins is rare as it is thermodynamically strongly disfavored. However, in "standard mechanism" protein inhibitors of serine proteinases, the P1 residue, which is highly exposed, becomes buried in the S1 specificity pocket of the enzyme. In many enzymes, such as Streptomyces griseus proteinase B (SGPB) the S1 pocket is hydrophobic. We measured the pH dependence of the association equilibrium constant for the interaction of SGPB with turkey ovomucoid third domain P1 mutants, Glu18 OMTKY3 and His18 OMTKY3. In order to eliminate the effects of other ionizable groups on the enzyme and the inhibitor, we divided these pH dependences by the pH dependence of the association equilibrium constant for the Gln18 OMTKY3 mutant. This yielded for Glu18, pKf (free inhibitor) of 4.46 +/- 0.05 and pKc (complex) of 8.74 +/- 0.06. For His18 the values are pKf 6.63 +/- 0.08 and pKc 4.31 +/- 0.07. At low pH values Glu18 variant is a relatively good inhibitor for SGPB. This may be biologically relevant. PMID- 7499198 TI - Characterization by electron paramagnetic resonance of the interactions of L arginine and L-thiocitrulline with the heme cofactor region of nitric oxide synthase. AB - Nitric oxide synthase (NOS) catalyzes sequential NADPH- and O2-dependent mono oxygenase reactions converting L-arginine to N omega-hydroxy-L-arginine and N omega-hydroxy-L-arginine to citrulline and nitric oxide. The homodimeric enzyme contains one heme/monomer, and that cofactor is thought to mediate both partial reactions. Here we show by electron paramagnetic resonance spectroscopy that binding of substrate L-arginine to neuronal NOS perturbs the heme cofactor binding pocket without directly interacting as a sixth axial heme ligand; heme iron is exclusively high spin. In contrast, binding of L-thiocitrulline, a NOS inhibitor, produces both high and low spin iron spectra; L-thiocitrulline sulfur is a sixth axial heme ligand in one, but not all, of the low spin forms. The high spin forms of the L-thiocitrulline NOS complex display a distortion in the opposite direction to that caused by L-arginine binding. The findings elucidate the binding interactions of L-arginine and L-thiocitrulline to neuronal NOS and demonstrate that each causes a unique perturbation to the heme cofactor pocket of NOS. PMID- 7499199 TI - Apolipoprotein A-I conformation in reconstituted discoidal lipoproteins varying in phospholipid and cholesterol content. AB - The effects of the size and cholesterol content on the conformation of apolipoprotein A-I (apoA-I) have been studied in reconstituted discoidal lipoproteins containing two apoA-I per particle (Lp2A-I). The immunoreactivity of a series of 13 epitopes distributed along the apoA-I sequence has been evaluated in Lp2A-I with a phospholipid/apoA-I molar ratio ranging from 31 to 156 and in Lp2A-I with constant phospholipids but varying in cholesterol content from 0 to 22 molecules. The results are compatible with a three domain structure in apoA-I in which the central domain is located between residues 99 and 143 and postulated to be a hinged domain that responds differentially to changes in phospholipid and cholesterol contents. Increasing the phospholipid content results in significant changes of epitope immunoreactivity throughout the N-terminal and central domains of apoA-I with fewer modifications in the C-terminal domain. In contrast, increasing Lp2A-I of two central epitopes, A11 (residues 99-132) and 5F6 (residues 118-148), and an extreme N-terminal epitope, 4H1 (residues 2-8). Interestingly, the effects of increasing cholesterol or phospholipids on these epitopes are opposite. This suggests a specific effect of cholesterol on the central domain tertiary structure between residues 99 and 143. Competition binding assays among pairs of antibodies binding to apoA-I on Lp2A-I are best explained by invoking inter- as well as intramolecular competitions. The specificity of the intermolecular competitions suggests an N to C termini arrangement of the two apoA-I molecules around the disc. Increasing the phospholipid content of Lp2A-I mainly increases the competitions between 3G10 and antibodies binding to most adjacent epitopes. Simultaneously as Lp2A-I enlarges, several of these antibodies also enhance the binding of 3G10. This has been interpreted as evidence of a structural rearrangement of apoA-I as a result of the size increase where the alpha-helix (residues 99-121) that contains the 3G10 epitope is increasingly interacting with lipids resulting in the enhanced expression of this epitope. The increasing interactions of apoA-I helices with lipids in the enlarging disc are compatible with previous reports of a greater apoA-I stability in the large discs. By contrast, cholesterol has limited but specific effects on antibody competitions and decreases the interaction of the N terminal domain with the domain containing 3G10, either by direct cholesterol protein interaction or by modification of the lipid phase packing. PMID- 7499200 TI - Immunochemical analysis of the structure of diphtheria toxin shows all three domains undergo structural changes at low pH. AB - Diphtheria toxin is a bacterial protein that undergoes a physiologically critical conformational change at low pH. This change involves a partial unfolding event forming a molten globule-like structure, which exposes hydrophobic regions and which allows the toxin to insert into, and translocate across, membranes. In this report, antibody binding was used to examine the regions of the toxin that undergo structural changes at low pH. Monoclonal antibodies specific to the catalytic (C), transmembrane (T), and receptor-binding (R) domains of diphtheria toxin were prepared and isolated. In addition, the binding of anti-peptide antibodies raised against peptides in the C and T domains to toxin was examined. Anti-C monoclonals and antipeptide antibodies were found to bind preferentially to low pH-treated toxin relative to native toxin. Anti-T and anti-R monoclonal binding ranged between preference for native toxin and preference for low pH treated toxin. These results suggest that the C domain becomes more exposed to solution at low pH, and that both the T and R domains of the B chain undergo major conformational changes at low pH. Based on these results, a model in which low pH induces several coordinated changes in intra- and inter-domain interactions is suggested. The participation of the R domain in these changes is of particular significance because it suggests that the R domain plays a more important role in low pH-induced changes than previously realized. PMID- 7499201 TI - Immunochemical analysis shows all three domains of diphtheria toxin penetrate across model membranes. AB - Diphtheria toxin undergoes membrane insertion and translocation across membranes when exposed to low pH. In this study, the translocation of the toxin has been investigated by the binding of antibodies to two preparations of model membrane inserted toxin. In one preparation, toxin was added externally to model membrane vesicles and then inserted by exposure to low pH. In the other preparation, toxin was entrapped in the vesicles at neutral pH, and then inserted by decreasing pH. At neutral pH, externally added antibodies could not bind to entrapped toxin, although they could bind to externally added native toxin. However, after low pH exposure, antibodies against all three toxin domains (catalytic (C), transmembrane (T), and receptor-binding (R)) could bind to entrapped toxin, and also to externally added membrane-inserted toxin. The binding to the entrapped toxin shows that all three domains of the toxin translocate to the trans face of the membrane after exposure to low pH. The observation that antibodies bind to both external and entrapped preparations of toxin after low pH exposure shows that toxin inserts in a mixed orientation. A difference in antibody binding to low pH-treated toxin in which the C domain is folded (Lr' conformation) or unfolded (Lr" conformation) was also observed. An increase in antibody binding to C and T domains in the Lr" conformation relative to binding to the Lr' conformation was found for entrapped toxin, suggesting that more of the C and T domains translocate across the bilayer in the Lr" conformation. These results suggest all three toxin domains insert in the membrane bilayer and participate in translocation in vitro. The C and R domains lack classical transmembrane hydrophobic sequences. However, they possess sequences that have the potential to form membrane-inserting beta-sheets. PMID- 7499202 TI - Changing stereochemistry for a metabolic pathway in vivo. Experiments with the peroxisomal beta-oxidation in yeast. AB - The biosphere is inherently built of chiral molecules, and once their metabolism is established, the stereochemical course of the reactions involved is seen to remain highly conserved. However, by replacing the yeast peroxisomal multifunctional enzyme (MFE), which catalyzes the second and third reactions of beta-oxidation of fatty acids via D-3-hydroxyacyl-CoA intermediates, with rat peroxisomal MFE, which catalyzes the same reactions via L-3-hydroxy intermediates, it was possible to change the chiralities of the intermediates in a major metabolic pathway in vivo. Both stereochemical alternatives allowed the yeast cells to grow on oleic acid, implying that when the beta-oxidation pathways evolved, the overall function was the determining factor for the acquisition of MFEs and not the stereospecificities of the reactions themselves. PMID- 7499203 TI - The structure of copper-nitrite reductase from Achromobacter cycloclastes at five pH values, with NO2- bound and with type II copper depleted. AB - High resolution x-ray crystallographic structures of nitrite reductase from Achromobacter cycloclastes, undertaken in order to understand the pH optimum of the reaction with nitrite, show that at pH 5.0, 5.4, 6.0, 6.2, and 6.8, no significant changes occur, other than in the occupancy of the type II copper at the active site. An extensive network of hydrogen bonds, both within and between subunits of the trimer, maintains the rigidity of the protein structure. A water occupies a site approximately 1.5 A from the site of the type II copper in the structure of the type II copper-depleted structure (at pH 5.4), again with no other significant changes in structure. In nitrite-soaked crystals, nitrite binds via its oxygens to the type II copper and replaces the water normally bound to the type II copper. The active-site cavity of the protein is distinctly hydrophobic on one side and hydrophilic on the other, providing a possible path for diffusion of the product NO. Asp-98 exhibits thermal parameter values higher than its surroundings, suggesting a role in shuttling the two protons necessary for the overall reaction. The strong structural homology with cupredoxins is described. PMID- 7499204 TI - Role of acidic residues in the interaction of NADPH-cytochrome P450 oxidoreductase with cytochrome P450 and cytochrome c. AB - Site-directed mutagenesis of the acidic clusters 207Asp-Asp-Asp209 and 213Glu-Glu Asp215 of NADPH-cytochrome P450 oxidoreductase demonstrates that both cytochrome c and cytochrome P450 interact with this region; however, the sites and mechanisms of interaction of the two substrates are clearly distinct. Substitutions in the first acidic cluster did not affect cytochrome c or ferricyanide reductase activity, but substitution of asparagine for aspartate at position 208 reduced cytochrome P450-dependent benzphetamine N-demethylase activity by 63% with no effect on KP450m or KNADPHm. Substitutions in the second acidic cluster affected cytochrome c reduction but not benzphetamine N demethylase or ferricyanide reductase activity. The E213Q enzyme exhibited a 59% reduction in cytochrome c reductase activity and a 47% reduction in KCyt cm under standard conditions (x0.27 M potassium phosphate, pH 7.7), as well as a decreased KCyt cm at every ionic strength and a shift of the salt dependence of cytochrome c reductase activity toward lower ionic strengths. The E214Q substitution did not affect cytochrome c reductase activity under standard conditions, but shifted the salt dependence of cytochrome c reductase activity toward higher ionic strengths. Measurements of the effect of ionic strength on steady-state kinetic properties indicated that increasing ionic strength destabilized the reductase-cytochrome c3+ ground state and reductase-cytochrome c transition state complexes for the wild-type, E213Q, and E214Q enzymes, suggesting the presence of electrostatic interactions involving Glu213 and Glu214 as well as additional residues outside this region. The ionic strength dependence of kcat/KCyt cm for the wild-type and E214Q enzymes is consistent with the presence of charge-pairing interactions in the transition state and removal of a weak ionic interaction in the reductase cytochrome c transition-state complex by the E214Q substitution. The ionic strength dependence of the E213Q enzyme, however, is not consistent with a simple electrostatic model. Effects of ionic strength on kinetic properties of E213Q suggest that substitution of glutamine stabilizes the reductase-cytochrome c3+ ground-state complex, leading to a net increase in activation energy and decrease in kcat. Glu213 is also involved in a repulsive interaction with cytochrome c3+. Cytochrome c2+ Ki for the wild-type enzyme was 82.4 microM at 118 mM ionic strength and 10.8 microM at 749 mM ionic strength; similar values were observed for the E214Q enzyme. Cytochrome c Ki for the E213Q enzyme was 17.6 microM at 118 mM and 15.7 microM at 749 mM ionic strength, consistent with removal of an electrostatic repulsion between the reductase and cytochrome c2+. PMID- 7499205 TI - Insulin-like growth factor (IGF)-binding protein-3 (IGFBP-3) functions as an IGF reversible inhibitor of IGFBP-4 proteolysis. AB - Previous studies have shown that insulin-like growth factor (IGF)-binding protein 4 (IGFBP-4) is degraded only in the presence of exogenous IGFs; however, we found that cation-dependent proteinase activity present in conditioned medium of MC3T3 E1 osteoblasts degrades 125I-recombinant human (rh)IGFBP-4 in the absence of IGFs. Addition of IGF-I, IGF-II, or insulin to conditioned medium had little affect on 125I-rhIGFBP-4 proteolysis, while extraction of IGFs resulted in only a approximately 10% reduction in proteinase activity. Since factors other than IGFs appeared to be involved in regulating IGFBP-4 proteolysis, we hypothesized that IGFBP-3, an IGFBP produced by many cell lines, but not MC3T3-E1 cells, might function as an inhibitor of IGFBP-4 proteolysis. Addition of rhIGFBP-3 to conditioned media inhibited 125I-rhIGFBP-4 proteolysis by 90%, while IGF-I and IGF-II reversed the inhibitory effects of rhIGFBP-3 in a dose-dependent manner. 125I-rhIGFBP-4 proteolysis was not inhibited by N-terminal rhIGFBP-3 fragments that bind IGFs, but was inhibited by two synthetic peptides corresponding to sequences contained in the mid-region or C-terminal region of IGFBP-3. Both inhibitory peptides contain highly basic, putative heparin-binding domains and heparin partially reversed the inhibitory effects of rhIGFBP-3 on 125I-rhIGFBP-4 proteolysis. These data demonstrate that rhIGFBP-3 inhibits IGFBP-4-degrading proteinase activity and binding of IGFs or glycosaminoglycans to IGFBP-3 may induce conformational changes in the binding protein, causing disinhibition of the proteinase. PMID- 7499206 TI - PD 098059 is a specific inhibitor of the activation of mitogen-activated protein kinase kinase in vitro and in vivo. AB - PD 098059 has been shown previously to inhibit the dephosphorylated form of mitogen-activated protein kinase kinase-1 (MAPKK1) and a mutant MAPKK1(S217E,S221E), which has low levels of constitutive activity (Dudley, D. T., Pang, L., Decker, S. J., Bridges, A. J., and Saltiel, A. R. (1995) Proc. Natl. Acad. Sci. U.S.A. 92, 7686-7689). Here we report that PD 098059 does not inhibit Raf-activated MAPKK1 but that it prevents the activation of MAPKK1 by Raf or MEK kinase in vitro at concentrations (IC50 = 2-7 microM) similar to those concentrations that inhibit dephosphorylated MAPKK1 or MAPKK1(S217E,S221E). PD 098059 inhibited the activation of MAPKK2 by Raf with a much higher IC50 value (50 microM) and did not inhibit the phosphorylation of other Raf or MEK kinase substrates, indicating that it exerts its effect by binding to the inactive form of MAPKK1. PD 098059 also acts as a specific inhibitor of the activation of MAPKK in Swiss 3T3 cells, suppressing by 80-90% its activation by a variety of agonists. The high degree of specificity of PD 098059 in vitro and in vivo is indicated by its failure to inhibit 18 protein Ser/Thr kinases (including two other MAPKK homologues) in vitro by its failure to inhibit the in vivo activation of MAPKK and MAP kinase homologues that participate in stress and interleukin-1 stimulated kinase cascades in KB and PC12 cells, and by lack of inhibition of the activation of p70 S6 kinase by insulin or epidermal growth factor in Swiss 3T3 cells. PD 098059 (50 microM) inhibited the activation of p42MAPK and isoforms of MAP kinase-activated protein kinase-1 in Swiss 3T3 cells, but the extent of inhibition depended on how potently c-Raf and MAPKK were activated by any particular agonist and demonstrated the enormous amplification potential of this kinase cascade. PD 098059 not only failed to inhibit the activation of Raf by platelet-derived growth factor, serum, insulin, and phorbol esters in Swiss 3T3 cells but actually enhanced Raf activity. The rate of activation of Raf by platelet-derived growth factor was increased 3-fold, and the subsequent inactivation that occurred after 10 min was prevented. These results indicate that the activation of Raf is suppressed and that its inactivation is accelerated by a downstream component(s) of the MAP kinase pathway. PMID- 7499207 TI - The binding of type I collagen to lymphocyte function-associated antigen (LFA) 1 integrin triggers the respiratory burst of human polymorphonuclear neutrophils. Role of calcium signaling and tyrosine phosphorylation of LFA 1. AB - Monoclonal antibodies to the alpha L beta 2 integrin inhibit the binding of type I collagen to PMN (polymorphonuclear neutrophil leukocytes) as well as the subsequent stimulation of superoxide production and enzyme secretion-elicited by this collagen. Pepsinized collagen still binds PMN but no longer stimulates them. The I domain of the alpha chain of the integrin is involved in the binding. Two sequences of the alpha 1(I) polypeptide chain of collagen participate in the process. Experiments of competitive inhibition by synthetic peptides showed that the sequence RGD (915-917) is used for binding to the cells and DGGRYY (1034 1039) serves to stimulate PMN. Experiments of radioactive labeling of the cells and affinity chromatography on Sepharose-collagen confirmed the presence in PMN extracts of two proteins, 95 and 185 kDa, respectively, corresponding to the molecular weights of the beta 2 and alpha L chains of the integrin and recognized by their specific monoclonal antibodies. The transduction pathways depending on the alpha L beta 2 integrin do not involve a G protein (ruled out by the use of cholera and pertussis toxins), whereas the cytoskeleton was found to participate in the process, as evidenced by inhibition by cytochalasin B. After collagen stimulation, cytoplasmic inositol trisphosphate and calcium ion increased sharply for less than 2 min. The use of the inhibitors staurosporine and calphostin C demonstrated that protein kinase C was involved. Evaluation of the activity of this enzyme showed that, upon stimulation of PMN with collagen I, it was translocated to plasma membrane. Acrylamide gel electrophoresis of the protein bands corresponding to the integrin alpha L beta 2, followed by immunoblotting using monoclonal antibodies to phosphotyrosine, permitted us to demonstrate that, prior to stimulation by type I collagen, there was no phosphorylation, whereas after stimulation, both alpha L and beta 2 chains were stained by anti phosphotyrosine antibodies. The adhesion of PMN to pepsinized type I collagen triggered tyrosine phosphorylation of the beta 2 chain of the integrin, without stimulating O2-. production by these cells, whereas their stimulation by complete type I collagen induced the tyrosine phosphorylation of both alpha L and beta 2 subunits. The tyrosine phosphorylation of both integrin subunits during transduction of stimuli is a heretofore undescribed phenomenon that may correspond to a new system of transmembrane communication. PMID- 7499208 TI - Association of activated phosphatidylinositol 3-kinase with p120cbl in antigen receptor-ligated B cells. AB - A 120-kDa protein that is tyrosine-phosphorylated upon antigen receptor ligation in B lymphocytes has been identified as the product of the c-cbl protooncogene. Tyrosine phosphorylation of Cbl depends on the efficient association of membrane immunoglobulin heavy chains with the Ig alpha/beta heterodimer but is unimpaired in splenic B cells from the Xid mouse. Cross-linking of membrane IgM and membrane IgG, but not of CD40, leads to the tyrosine phosphorylation of Cbl. In receptor ligated B lymphocytes, p120cbl associates with an 85-kDa protein that has been identified as the 85-kDa subunit of phosphatidylinositol 3-kinase. PMID- 7499209 TI - Mitochondrial calcium uptake from physiological-type pulses of calcium. A description of the rapid uptake mode. AB - A controversy in the field of bioenergetics has been whether mitochondria are capable of sequestering enough Ca2+ from cytosolic Ca2+ pulses to raise their intramitochondrial free Ca2+ level ([Ca2+]m). This is significant because an increase in [Ca2+]m has been linked to an increase in cellular metabolic rate through various mechanisms. To resolve this question, we exposed isolated liver mitochondria to physiological type pulses of Ca2+ produced using a pulse generating system (Sparagna, G. C., Gunter, K. K., and Gunter, T. E. (1994) Anal. Biochem. 219, 96-103). We then measured the resulting mitochondrial Ca2+ uptake. The uniporter was previously thought to be the only specific Ca2+ uptake mechanism in mitochondria. Our studies have uncovered an additional uptake mechanism, the rapid mode of uptake or RaM, which functions at the beginning of each pulse and allows mitochondria to sequester a considerable amount of Ca2+ from short pulses. We have shown that the RaM is reset by decreasing the [Ca2+] between pulses for a very short time, making this uptake mode ideally suited for Ca2+ sequestration from Ca2+ pulse sequences. With rapid Ca2+ uptake occurring at the beginning of each pulse, liver mitochondria may be able to sequester sufficient Ca2+ from a short sequence of pulses to activate the cellular metabolic rate. PMID- 7499210 TI - The reduction of membrane-bound dopamine beta-monooxygenase in resealed chromaffin granule ghosts. Is intragranular ascorbic acid a mediator for extragranular reducing equivalents? AB - The role of internal and external reductants in the dopamine beta-monooxygenase (D beta M)-catalyzed conversion of dopamine to norepinephrine has been investigated in resealed chromaffin granule ghosts. The rate of norepinephrine production was not affected by the exclusion of internal ascorbate. The omission of ascorbate from the external medium drastically reduced the norepinephrine production without affecting the net rate of dopamine uptake. In the presence of the external reductant, the internal ascorbate levels were constant throughout the incubation period. The rate of norepinephrine production was not affected when ghosts were resealed to contain the D beta M reduction site inhibitor, imino D-glucoascorbate. Ghosts incubated with external imino-D-glucoascorbate reduced the norepinephrine production. The weak D beta M reductant, 6-amino-L-ascorbic acid, was found to be a good external reductant for granule ghosts. The outcome of the above experiments was not altered when dopamine was replaced with the reductively inactive D beta M substrate, tyramine. These results and the known topology of membrane-bound D beta M disfavor the direct reduction of the enzyme by the external reductant. Our observations are consistent with the hypothesis that external ascorbate is the sole source of reducing equivalents for D beta M monooxygenation and that internal soluble ascorbate (or dopamine) may not directly reduce or mediate the reduction of membrane-bound D beta M in resealed granule ghosts. PMID- 7499211 TI - Expression of type V adenylyl cyclase is required for epidermal growth factor mediated stimulation of cAMP accumulation. AB - Previously, this laboratory has demonstrated that epidermal growth factor (EGF) increases adenylyl cyclase activity in cardiac membranes and elevates cAMP accumulation in hearts and cardiac myocytes. Since EGF does not increase cAMP accumulation in all tissues, we investigated the possibility that the expression of a specific isoform of adenylyl cyclase (AC) was necessary to observe EGF elicited stimulation of cAMP accumulation. HEK 293 cells were transfected with different isoforms of AC, and the ability of EGF to increase AC activity as well as elevate cAMP accumulation was determined. In cells transfected with AC I, II, V, and VI cDNAs, neither the expression nor the amount of the two isoforms of Gs alpha (45 and 52 kDa) were altered. Similarly, EGF-elicited phosphorylation of cellular proteins on tyrosine residues in various transfectants was unaltered. However, EGF increased AC activity and elevated cAMP accumulation only in cells expressing the rat and canine ACV. EGF did not alter either AC activity or cAMP accumulation in cells overexpressing types I, II, and VI isozymes. As assessed by the ability of an anti-Gs alpha antibody to obliterate the effect, stimulation of AC activity in AC V transfectants involved the participation of Gs alpha, a finding consistent with previous data concerning EGF effects on cardiac AC (Nair, B. G., Parikh, B., Milligan, G., and Patel, T. B. (1990) J. Biol. Chem. 265, 21317-21322). Thus we conclude that the expression of AC V isoform confers specificity to the ability of EGF to stimulate AC activity. PMID- 7499212 TI - TOR mutations confer rapamycin resistance by preventing interaction with FKBP12 rapamycin. AB - The antifungal, immunosuppressive compound rapamycin arrests the cell cycle in G1 in both yeast cells and T-lymphocytes. Previous genetic studies in yeast identified mutations in three genes, FPR1 (FKBP12), TOR1, and TOR2, which confer rapamycin resistance, and genetic findings implicated the TOR proteins as direct targets of FKBP12-rapamycin. Consistent with this model, we find that modulating TOR1 and TOR2 expression alters rapamycin sensitivity. We describe several TOR2 mutations that confer rapamycin resistance. These mutations prevent FKBP12 rapamycin binding to TOR2, as assayed with the two-hybrid system. We find that TOR1 and the mammalian TOR homologue (mTOR) also bind FKBP12-rapamycin, and mutations corresponding to those in TOR2 similarly block FKBP12-rapamycin binding. We demonstrate that FKBP12 prolyl isomerase activity is not required for FKBP12-rapamycin binding to TOR and that a composite protein-drug surface contacts the TOR proteins. These studies confirm that the TOR proteins are direct targets of FKBP12-rapamycin, reveal that drug-resistant mutations prevent this association, and define structural features of these complexes. PMID- 7499213 TI - Identification of a novel repressive element in the proximal lck promoter. AB - The T-cell-specific protooncogene lck, a src-related tyrosine kinase, is under the control of two promoters that give rise to transcripts differing only in their 5'-untranslated regions. The distal promoter is transcriptionally active in both peripheral and thymic T-cells, whereas expression of the proximal promoter is highest in developing thymocytes. The proximal promoter has also been shown to be selectively activated in a number of colon carcinoma cell lines. Approximately 570 base pairs of proximal promoter sequence is required for expression in both T cells and colon carcinoma cell lines. Protein binding studies were initiated with an oligonucleotide homologous to a region that, when deleted, causes an increase in promoter activity in transgenic animals. Two proteins with approximate molecular masses of 35 and 75 kDa were found to bind to this region as determined by UV cross-linking studies. Absence of specific protein binding is correlated with a high level of proximal promoter expression. Competitive gel retardation analysis identified a 9-base pair binding site within the proximal lck promoter that is necessary for repression of transcription in cells that contain specific binding activity. Mutants of this binding site do not repress transcription. Repression does not occur in a cell line that expresses lck and lacks this activity. These data support the hypothesis that activation of lck transcription in colon carcinoma is due, at least in part, to the loss of a transcriptional repressor. PMID- 7499214 TI - Characterization of a sulfotransferase from human airways responsible for the 3-O sulfation of terminal galactose in N-acetyllactosamine-containing mucin carbohydrate chains. AB - A galactose 3-O-sulfotransferase activity able to transfer a sulfate group from adenosine 3'-phosphate 5'-phosphosulfate to methyl galactosides or terminal N acetyllactosamine-containing carbohydrate chains from human respiratory mucins was characterized in microsomal fractions prepared from human respiratory mucosa. The reaction products, methyl alpha- or beta-galactose 3-sulfate and three oligosaccharide alditols containing the sequence HSO3-3Gal beta 1-4GlcNAc beta 1 6GalNAc-itol were identified by high performance anion-exchange chromatography. Using methyl beta-galactoside as a substrate, the optimum activity was obtained with 0.1% Triton X-100, 30 mM NaF, 20 mM Mn2+, and 10 mM AMP in a 30 mM 2-(N morpholino)ethanesulfonic acid buffer at pH 6.1. The apparent Km for methyl beta galactoside and for adenosine 3'-phosphate 5'-phosphosulfate were observed at 0.69 x 10(-3) M and at 4 x 10(-6) M respectively. This sulfotransferase is different from that responsible for sulfatide synthesis. PMID- 7499216 TI - Studies on the dissociation of cryptomonad biliproteins. AB - The spectroscopic properties of two biliproteins, phycocyanin 645 and phycoerythrin 566, have been studied by treating the proteins with two different agents, NaSCN at pH 6.0, or pH 4.0 without NaSCN. For phycoerythrin 566, treatment with NaSCN revealed that the visible CD spectrum of its chromophores was separated into a pair of different spectra, and each of these spectra was observed as a negative and one or more positive bands. For phycocyanin 645, two negative CD bands have been observed previously, together with two or more positive bands, in the dimer (alpha 2 beta 2) state, and NaSCN treatment caused elimination of one of these negative bands. The dimer was stable at pH 6.0, but at pH 4.0 the spectra of phycocyanin 645 had one less negative band than those at pH 6.0. Chromatography demonstrated that phycocyanin 645 was a monomer (alpha beta) at pH 4.0. Monomers of cryptomonad biliproteins have never been observed before. Excitation at 514 nm, in picosecond time-resolved fluorescence studies, produced lifetimes of 11.0 and 45.2 ps for dimers and monomers, respectively. Excitation at 566 nm yielded times of 1.38 and 1.24 ps, for dimers and monomers, respectively. CD in the far UV showed that monomers and dimers had very similar secondary structures. These results have been used to test an hypothesis that proposed two types of exciton splitting among the chromophores of phycocyanin 645, and perhaps phycoerythrin 566 could also have this chromophore organization. PMID- 7499215 TI - A direct demonstration of the catalytic action of monodehydroascorbate reductase by pulse radiolysis. AB - To elucidate the catalytic mechanism of monodehydroascorbate (MDA) reductase from cucumber, its interaction with MDA radical was investigated by the use of pulse radiolysis. When approximately equimolar MDA radical to the fully reduced MDA reductase was generated, the fully reduced enzyme reacted first with MDA radical to form the red semiquinone, and the semiquinone further reacted with MDA radical to form the oxidized enzyme. At a low ratio (< 20) of MDA radical to enzyme concentration, the fully reduced enzyme reacted quantitatively with MDA radical to form the semiquinone with a second-order rate constant of 2.6 x 10(8) M-1 s-1 at pH 7.4. At excess MDA radical to enzyme concentration, a similar rate constant was obtained from the decay of MDA radical. These results suggest that the reaction of the semiquinone with MDA radical occurs at the same rate or rate limiting step of the oxidation of the fully reduced enzyme by MDA radical. The rate constants decreased with an increase in NaCl concentration, suggesting that the localization of cationic groups of amino acid residue near the active site may provide electrostatic guidance to the anionic substrate of MDA radical. PMID- 7499217 TI - Fluorescence resonance energy transfer reveals interleukin (IL)-1-dependent aggregation of IL-1 type I receptors that correlates with receptor activation. AB - Fluorescence resonance energy transfer (FRET) was used to investigate whether interleukin-1 (IL-1) causes the aggregation of IL-1 type I receptors (IL-1 RI) at the cell surface. For these experiments, a noncompetitive anti-IL1 RI monoclonal antibody, M5, was labeled separately with a donor probe, fluorescein isothiocyanate, or with an acceptor carbocyanine probe, Cy3. Donor-labeled M5 and acceptor-labeled M5 were simultaneously bound to transfected mouse IL-1 RI on either C-127 mouse mammary carcinoma cells or on Chinese hamster ovary (CHO)-K1 cells, and the ratio of acceptor emission at 590 nm to donor emission at 525 nm (excitation at 488 and 514 nm) was monitored with flow cytometry as an indicator of FRET. Addition of a saturating concentration of human IL-1 alpha at 22 degrees C causes a time-dependent increase in FRET for both cell lines that indicates IL 1-dependent self-association of IL-1 RI. Binding of the IL-1 receptor antagonist at 22 degrees C causes little or no FRET for both cell lines, indicating a correlation between receptor aggregation and the ability of the ligand to stimulate a functional response. When donor-labeled and acceptor-labeled Fab fragments of M5 are used to monitor FRET, IL-1 alpha causes efficient energy transfer in the CHO-K1 cells at 22 degrees C, but not at 4 degrees C. In contrast, IL-1 alpha causes much less FRET at 22 degrees C in C-127 cells when the M5 Fab fragments are used instead of the intact bivalent M5. In a striking parallel, IL-1 alpha-dependent activation of prostaglandin E2 production depends on the bivalent M5 antibody in the C-127 cells, but is independent of this monoclonal antibody in the CHO-K1 cells. These results provide a strong correlation between the ability of IL-1 to cause the aggregation of IL-1 RI and the stimulation of a functional response. PMID- 7499218 TI - Ligand-induced conformational alterations of the androgen receptor analyzed by limited trypsinization. Studies on the mechanism of antiandrogen action. AB - Limited proteolysis of in vitro produced human androgen receptor was used to probe the different conformations of the receptor after binding of androgens and several antiandrogens. The results provide evidence for five different conformations of the receptor, as detected by the formation of proteolysis resisting fragments: 1) an initial conformation of the unoccupied receptor not resisting proteolytic attack; and receptor conformations characterized by 2) a 35 kDa proteolysis resisting fragment spanning the ligand binding domain and part of the hinge region, obtained with most antagonists, and in an initial step after agonist binding; 3) a 29-kDa proteolysis resisting fragment spanning the ligand binding domain, obtained in the presence of agonists after an activation process; 4 and 5) 30- and 25-kDa fragments, derived from 2 and 3, but missing part of the C terminus, obtained with RU486 (RU486 has antiandrogenic properties, besides its effects as an antiprogestagen/antiglucocorticoid). Concomitantly with the change from 2 to 3 (and of 4 to 5 for RU486), dissociation of the 8 S complex of receptor with associated proteins occurred. With a mutant receptor (LNCaP cell mutation in C-terminal region), some antagonists activated transcription analogous to agonists, and induced the activated receptor conformation 3. A mutant lacking the C-terminal 12 amino acids bound RU486 but not androgens, and formed with RU486 conformation 5. These data imply that, after the initial rapid binding of ligand, androgens induce a conformational change of the receptor, a process that also involves release of associated proteins. RU486 induces an inappropriate conformation of the C-terminal end, similar as found for its effect on the progesterone receptor. In contrast, the other antiandrogens act at a different step in the mechanism of action: they do not induce an abnormal conformation, but act earlier and prevent a conformation change by stabilizing a complex with associated proteins. PMID- 7499219 TI - Angiotensin II stimulates system y+ and cationic amino acid transporter gene expression in cultured vascular smooth muscle cells. AB - The effect of angiotensin II (Ang II) on the transport of cationic amino acids has been examined in vascular smooth muscle cells (VSMC) isolated from rat aortae. Ang II stimulated the uptake rates of radiolabeled arginine and lysine in a time- and concentration-dependent manner. The stimulated arginine uptake could be blocked by pretreatments with cycloheximide and actinomycin D or co-treatment with valsartan, an antagonist specific for Ang II receptor subtype-1. The modulation by Ang II was bidirectional as the efflux of arginine was also stimulated, 5-fold over basal. Using reverse transcription-coupled polymerase chain reaction methodology, a partial cDNA with 94% sequence identity to that of cationic amino acid transporter subtype-1 (CAT-1) of mouse fibroblasts was obtained from VSMC. This sequence also exhibited 14 base changes compared with the sequence of ecotropic retrovirus receptor (ERR)/CAT-1 from rat hepatoma. Northern analyses with this partial CAT-1 cDNA and CAT-2 cDNA of mouse T lymphocytes showed that Ang II rapidly stimulated the expression of both CAT-1 and CAT-2 in VSMC. Both signals peaked at 2 h after exposure to Ang II. The CAT-1 signal decayed over the next 6 h to levels 3-fold above basal, which are maintained up until 24 h. The induced CAT-2 mRNA concentration also decayed rapidly but increased again between 16 and 24 h to levels comparable with those observed at 2 h. PMID- 7499221 TI - BiP binding sequences in antibodies. AB - During the process of folding and assembly of antibody molecules in the endoplasmic reticulum, immunoglobulin heavy and light chains associate transiently with BiP, a resident endoplasmic reticulum protein that is a member of the Hsp70 family of molecular chaperones. BiP is thought to recognize unfolded or unassembled polypeptides by binding extended sequences of approximately seven amino acids that include bulky hydrophobic residues not normally exposed on the surface of native proteins. We used a computer algorithm developed to predict BiP binding sites within protein primary sequences to identify sites within immunoglobulin chains that might mediate their association with BiP. Very few of the sequential heptapeptides in the heavy or light chain sequences were potential BiP binding sites. Analysis of the ability of synthetic heptapeptides corresponding to 24 potential sites in heavy chains to stimulate the ATPase activity of BiP indicated that at least half of them were authentic BiP binding sequences. These sequences were not confined to a single domain of the heavy chain but were distributed within both the VH and CH domains. Interestingly, when the BiP binding sequences were mapped onto the three-dimensional structure of the Fd antibody fragment, the majority involve residues that participate in contact sites between the heavy and light chains. Therefore, we suggest that in vivo BiP chaperones the folding and assembly of antibody molecules by binding to hydrophobic surface regions on the isolated immunoglobulin chains that subsequently participate in interchain contacts. PMID- 7499220 TI - Identification and characterization of an exercise-sensitive pool of glucose transporters in skeletal muscle. AB - Augmentation of glucose transport into skeletal muscle by GLUT4 translocation to the plasma and T-tubule membranes can be mediated independently by insulin and by contraction/exercise. Available data suggest that separable pools of intracellular GLUT4 respond to these two stimuli. To identify and characterize these pools, we fractionated skeletal muscle membranes in a discontinuous sucrose density gradient. Fractions of 32 and 36% sucrose exhibited the highest enrichment of GLUT4 and were independently responsive to insulin and exercise, respectively. The combination of the two stimuli depleted both GLUT4 fractions simultaneously. Both vesicle populations contained the gp160 aminopeptidase, whose expression had previously been shown to be specific to muscle and fat and restricted to GLUT4 vesicles in the latter tissue. In muscle, gp160 translocates exactly as does GLUT4 in response to insulin and exercise. The contraction- and insulin-sensitive GLUT4 pools also contained secretory component-associated membrane protein/glucose transporter vesicle triplet but not GLUT1 and caveolin. Immunoadsorption of the two pools followed by silver staining did not reveal any obvious difference in their major protein components. On the other hand, sedimentational analysis in sucrose velocity gradients revealed that the insulin sensitive GLUT4 vesicles had a larger sedimentation coefficient than the exercise sensitive vesicles. Thus, the separation of the two intracellular GLUT4 pools should be useful in dissecting what are likely to be different signal transduction pathways that mediate their translocation to the cell surface. PMID- 7499222 TI - Structural role of extracellular domain 1 of alpha-platelet-derived growth factor (PDGF) receptor for PDGF-AA and PDGF-BB binding. AB - The purpose of this study was to bacterially express, purify, and refold combinations of the extracellular immunoglobulin (Ig)-like domains (2-3, 1-3, and 1-5) of the human alpha-platelet-derived growth factor receptor (alpha PDGFR) to characterize molecular interactions with its ligand, platelet-derived growth factor (PDGF). The far UV circular dichroism spectroscopy of the alpha-PDGFR extracellular domains (ECDs) revealed a predominantly beta-sheet protein, with a structure consistent with folded Ig-like domains. The addition of PDGF-BB to these ECD types changed the conformation of all three types with a decrease in mean residue ellipticity in the following rank order: 1-5 = 1-3 > 2-3. In striking contrast, addition of PDGF-AA to these ECD types markedly changed the conformation of ECD 2-3, by an increased mean residue ellipticity but no changes were observed for ECDs 1-3 and 1-5. PDGF-AA bound to the immobilized ECD types 2 3, 1-3, and 1-5 at concentrations of 20, 11, and 7.5 nM, respectively. In contrast, PDGF-BB bound the ECD types 2-3, 1-3, and 1-5 at concentrations of 3, 3, and 2.2 nM, respectively. Scatchard analysis of binding studies using labeled ECDs indicated that PDGF-BB bound ECD 1-3 and ECD 2-3 with KD values of 74 and 72 nM, respectively. While, PDGF-AA bound ECD 1-3 and ECD 2-3 with KD values of 33 and 87 nM, respectively. Therefore, our results indicated that the loss of ECD 1 impaired the binding affinity of alpha PDGFR ECD 1-3 toward PDGF-AA without having a similar effect on PDGF-BB binding. Together all of our data suggest that ECD 1 is differentially required for proper orientation of PDGF-AA but not PDGF BB binding determinant within ECDs 2 and 3. PMID- 7499224 TI - Structure and topology of the influenza virus fusion peptide in lipid bilayers. AB - The secondary structure of a 20-amino acid length synthetic peptide corresponding to the N terminus of the second subunit of hemagglutinin (HA2) of influenza virus A/PR8/34 and its interaction with phospholipid bilayers are investigated using ESR, Fourier transform infrared (FTIR), and CD spectroscopy. N-terminal spin labeling of the peptide did not affect the secondary structure of the peptide either in solution or when bound to liposomes as revealed by FTIR and CD spectroscopy. ESR spectra show that the mobility of the labeled peptide is dramatically restricted in the presence of phosphatidylcholine liposomes, suggesting a strong binding to the lipid membranes. The N terminus of the peptide penetrates into the membrane and is located within the hydrophobic core. We find an oblique insertion of the peptide into the lipid bilayer with an angle of about 45 degrees between helix axis and membrane plane using FTIR spectroscopy. No gross changes of the peptide's orientation, motion, and secondary structure were observed between pH 7.4 and pH 5.0. A model of the insertion of the fusion sequence of HA2 into a lipid bilayer is presented taking into account recent investigations on the low pH conformation of HA2 (Bullough, P. A., Hughson, F. M., Skehel, J. J., and Wiley, D. C. (1994) Nature 371, 37-43). PMID- 7499223 TI - A single mutation of the neurokinin-2 (NK2) receptor prevents agonist-induced desensitization. Divergent conformational requirements for NK2 receptor signaling and agonist-induced desensitization in Xenopus oocytes. AB - Receptor activation and agonist-induced desensitization of the human neurokinin-2 (NK2) receptor expressed in Xenopus oocytes have been investigated. When neurokinin A (NKA) was applied repeatedly at 5-min intervals, the second and subsequent applications gave no responses. This desensitization was not observed with the specific agonists (Lys3, Gly8-R-gamma-lactam-Leu9)NKA(3-10) (GR64349) or (Nle10)-NKA(4-10). However, in the presence of the protein kinase inhibitor staurosporine, stimulation with GR64349 or (Nle10)-NKA(4-10) induced receptor desensitization. In contrast, the protein kinase C inhibitor Ro-31-8220 was not able to enhance GR64349-mediated desensitization. We created a mutation (F248S) in the third cytoplasmic loop of NK2 that impairs NKA-induced desensitization. In the presence of either staurosporine or Ro-31-8220, the mutant receptor was desensitized in response to NKA application but not to GR64349. Also, truncation mutants delta 62 and delta 87, lacking serine and threonine residues in the cytoplasmic COOH-terminal tail, were functionally active and were partially resistant to desensitization. These observations indicate that 1) there are different conformational requirements for NK2 receptor signalling and agonist induced desensitization, 2) the third intracellular loop and the cytoplasmic tail of NK2 are functional domains important for agonist-induced desensitization, and 3) some agonists at the NK2 receptor cause much more desensitization than others and suggest that this might result from phosphorylation by receptor-specific kinases and other non-identified protein kinases. PMID- 7499225 TI - A conserved region of c-Ha-Ras is required for efficient GTPase stimulation by GTPase activating protein but not neurofibromin. AB - The effector binding domain and the switch II region of c-Ha-Ras are necessary for p120GAP-stimulated GTP hydrolysis. We report a third region of c-Ha-Ras located within the alpha 3 helix (amino acids 101-103) which is also required for efficient p120GAP, but not neurofibromin-mediated hydrolysis. This highly conserved region of the Ras protein was investigated using an insertion-deletion mutant (Ras-100LIR104) originally characterized by Willumsen et al. (Willumsen, B. M., Adari, H., Zhang, K., Papageorge, A. G., Stone, J. C., McCormick, F., and Lowy, D. R (1989) in The Guanine Nucleotide Binding Proteins; Common Structural and Functional Properties (Bosch, L., Kraal, B., and Parmeggiani, A., eds) pp. 165-178, Plenum Press, New York). The 100LIR104 substitution did not alter the intrinsic hydrolytic rate of the protein. The p120GAP-stimulated hydrolysis of Ras-100LIR104, however, was decreased by 2-3-fold compared to wild type Ras. This decrease in p120GAP-stimulated hydrolysis was not due to its inability to physically associate with Ras-100LIR104. GTP (as determined by competitive binding assays). Surprisingly, neurofibromin-stimulated GTP hydrolysis was unaltered by the mutation. Finally, no differences were observed in the ability of either the p120GAP catalytic domain or the neurofibromin GRD to accelerate Ras 100LIR104 GTPase activity, indicating that the amino-terminal noncatalytic GAP region is critical for p120GAP-stimulated GTP hydrolysis. This is the first report of a Ras mutation which differentiates between p120GAP and neurofibromin activity. PMID- 7499226 TI - v-src induces prostaglandin synthase 2 gene expression by activation of the c-Jun N-terminal kinase and the c-Jun transcription factor. AB - A consensus cyclic AMP response element (CRE) in the murine prostaglandin synthase-2 (PGS2) promoter is essential for pgs2 gene expression induced by pp60v src, the v-src oncogene product. In this study, we investigate (i) the transcription factors active at the PGS2 "CRE site" in response to v-src activation and (ii) the signal transduction pathways by which pp60v-src activates these transcription factors. Transient transfection assays with pgs2 promoter/luciferase reporter chimeric genes suggest that c-Jun mediates v-src induced pgs2 gene expression. Antibody supershift experiments demonstrate that c Jun can participate in a complex with the pgs2 promoter CRE site. Moreover, in vitro immuno-complex assays demonstrate that pp60v-src expression strongly activates c-Jun N-terminal kinase (JNK1) enzyme activity. Serines 63 and 73, the sites of c-Jun phosphorylation by JNK, are essential for v-src-induced, pgs2 promoter-mediated luciferase expression. Cotransfection studies with plasmids expressing wild-type JNK, dominant-negative JNK, and dominant-negative MEKK-1 confirm that activation of the Ras/MEKK-1/JNK/c-Jun pathway is required for v-src induced pgs2 gene expression. Overexpression of either wild-type ERK-1 or ERK-2 proteins also potentiate v-src-mediated luciferase expression driven by the pgs2 promoter, and expression of dominant-negative mutants of ERK-1, ERK-2, or Raf-1 attenuate this response. Thus, in response to v-src expression, a Ras/MEKK-1/JNK signal transduction pathway activating c-Jun and a Ras/Raf-1/ERK pathway converge to mediate pgs2 gene expression via the CRE site in the pgs2 promoter. PMID- 7499227 TI - Identification of proximal sequence element nucleotides contributing to the differential expression of variant U4 small nuclear RNA genes. AB - The two U4 genes in the chicken genome code for distinct sequence variants of U4 small nuclear RNA that are differentially expressed during development. Whereas U4B RNA is constitutively expressed, U4X RNA is specifically down-regulated relative to U4B in a tissue-specific manner during development. To investigate mechanisms controlling the differential expression of the U4B and U4X genes, chimeric U4 genes were constructed and their transcriptional activities assayed by injection into Xenopus oocytes or by transfection of CV-1 cells. The proximal regulatory region of the U4B gene and the enhancers of both the U4B and U4X genes functioned efficiently in each expression system. However, the proximal region of the U4X gene was inactive. To localize and identify the responsible nucleotides, reciprocal point mutations were introduced into the U4X and U4B proximal regulatory regions. The results indicate that the U4X gene contains a suboptimal proximal sequence element, and that this results primarily from the identities of the nucleotides at positions -61 and -57 relative to the transcription start site. PMID- 7499228 TI - Unusual anionic N-linked oligosaccharides from bovine lung. AB - We previously described a diverse family of sulfated anionic N-linked oligosaccharides released by peptide: N-glycosidase F (PNGaseF) from calf pulmonary artery endothelial (CPAE) cells (Roux, L., Holoyda, S., Sundblad, G., Freeze, H.H., and Varki, A. (1988) J. Biol. Chem. 263, 8879-8889). Since a major fraction of the intact lung consists of endothelial cells, we reasoned that bovine lung might be a rich source of similar molecules. Total N-linked oligosaccharides from bovine lung acetone powder were released by PNGaseF, labeled by [3H]NaBH4 reduction, and the anionic fractions were studied with a variety of techniques. The sugar chains with lesser negative charge (designated Class I) share several properties of conventional multiantennary complex-type chains. However, unlike the case with CPAE cells, sialic acids account only for a minority of the anionic properties and only a small proportion carry sulfate esters. A variety of different treatments indicate that most of the unexplained negative charge is due to multiple carboxylic acid groups. Resistance to beta glucuronidase and alpha-iduronidase suggests that these may be previously undescribed modifications of mammalian oligosaccharides. The most highly charged N-linked chains (designated Class II) are more similar in general structure to the corresponding ones from CPAE cells, although relatively more abundant. Their high charge is primarily due to chondroitin sulfate, heparin/heparan sulfate, or keratan sulfate glycosaminoglycan chains. Sequential digestion studies suggest that a significant proportion of these molecules have more than one type of glycosaminoglycan chain associated with them. Compositional analysis indicates the presence of xylose residues in Class II, but not Class I molecules. However, unlike the case with conventional glycosaminoglycans, these residues are not at the reducing terminus. Most previously reported structures of complex-type N linked oligosaccharides are derived from the glycoproteins of blood cells, plasma, or the secretions of cultured mammalian cells. This library of N-linked oligosaccharides from an intact mammalian organ (lung) contains a high proportion of novel anionic sugar chains whose structures are different from conventional complex-type sialylated chains and only partially related to those from CPAE cells. Further exploration of the N-linked chains of intact mammalian tissues seems warranted. PMID- 7499229 TI - Inhibition of lipopolysaccharide biosynthesis and cell growth following inactivation of the kdtA gene in Escherichia coli. AB - The enzyme 3-deoxy-D-manno-octulosonic acid (Kdo) transferase is encoded by the kdtA gene in Escherichia coli. The enzyme is a single polypeptide that catalyzes the transfer of two Kdo residues to a tetraacyldisaccharide-1,4'-bisphosphate precursor of lipid A, designated lipid IVA (Belunis, C.J., and Raetz, C.R.H. (1992) J. Biol. Chem. 267, 9988-9997). To determine if Kdo transfer to lipid IVA is required for growth, we constructed a strain of E. coli with a chromosomal kdtA::kan insertion mutation. In mutants carrying the kdtA::kan allele on the chromosome, cell growth and Kdo transferase activity were dependent upon a copy of the intact kdtA gene on a plasmid. When the kdtA-bearing plasmid was itself temperature sensitive for replication, the growth of these strains was inhibited after several hours at 44 degrees C, and Kdo transferase activity in extracts became undetectable. Concomitantly, the cells accumulated massive amounts of lipid IVA, the precursor of (Kdo)2-lipid IVA. The kdtA::kan mutation could also be complemented by hybrid plasmids bearing the gseA gene of Chlamydia trachomatis. gseA specifies a distinct Kdo transferase that adds three Kdo moieties to lipid IVA. Lipopolysaccharide from E. coli kdtA::kan constructs complemented by gseA reacts strongly with antibodies directed against the genus specific epitope of Chlamydia, whereas lipopolysaccharide from parental E. coli K 12 does not. Our studies prove that Kdo attachment during lipid A biosynthesis is essential for cell growth and accounts for the conditional lethality associated with mutations in Kdo biosynthesis. PMID- 7499230 TI - Characterization of the mitotic specific phosphorylation site of histone H1. Absence of a consensus sequence for the p34cdc2/cyclin B kinase. AB - 32P-Labeled histone H1 was isolated from synchronized Chinese hamster (line CHO) cells, subjected to tryptic digestion, and fractionated into 15 phosphopeptides by high performance liquid chromatography. These phosphopeptides were grouped into five classes having different cell cycle phosphorylation kinetics: 1) peptides reaching a maximum phosphorylation rate in S and then declining in G2 and M, 2) peptides reaching a maximum phosphorylation rate in G2 and then remaining constant or declining in M, 3) peptides with increasing phosphorylation throughout S and G2 and reaching a maximum in M, 4) one peptide that was phosphorylated only in M, and 5) peptides that had low levels of phosphorylation that remained constant throughout the cell cycle. Amino acid analysis and sequencing demonstrated that the mitotic specific H1 phosphopeptide was the 16 amino acid, N-terminal, tryptic peptide Ac-SETAPAAPAAAPPAEK of the H1-1 class. This peptide, which is phosphorylated on both the Ser and Thr, does not contain the consensus sequence (S/T)PXZ (where X is any amino acid and Z is a basic amino acid). This sequence is thought to be required by the p34cdc2/cyclin B kinase that has maximum phosphorylating activity in mitosis. These data indicate that this kinase either does not have an obligatory requirement for the consensus sequence in vivo as generally believed or that it is not the enzyme responsible for the mitotic specific H1 phosphorylation. PMID- 7499231 TI - Mutation of serine 90 to glutamic acid mimics phosphorylation of bovine prolactin. AB - Phosphorylated prolactin has been identified and isolated from bovine pituitaries. The biological activity of this phosphoprotein is severely reduced in comparison with nonphosphorylated prolactin. The sites of phosphorylation are serines 26, 34, and 90, and the stoichiometry is 1:1:10, respectively. In this report, the phosphoserine residues have been individually replaced with glutamic acid in recombinant methionyl bovine prolactins in order to mimic phosphorylation at each site. Substitution of glutamic acid for serine at positions 26, 34, and 90 reduced protein helical contents by 10, 6, and 14%, respectively. UV absorbances for S26E and S34E bovine prolactins were blue-shifted, similar to the biological isolates of phosphorylated bovine prolactin, but the biological activities of the S26E and S34E mutants (ED50 values of 16.3 and 18.8 pM, respectively) were similar to that of wild-type prolactin (ED50 value of 18.6 pM) in the Nb2 rat lymphoma assay. S90E bovine prolactin had the greatest reduction in helical content but showed similar UV and fluorescent spectra to the wild-type bovine prolactin. The biological activity of S90E bovine prolactin (ED50 value of 672 pM) was reduced to an activity similar to that of phosphorylated bovine prolactin. The data indicate that the phosphorylation of serine 90 is responsible for the reduction in biological activity. PMID- 7499232 TI - Exposure of B-lineage lymphoid cells to low energy electromagnetic fields stimulates Lyn kinase. AB - Here, we present evidence that exposure of B-lineage lymphoid cells to low energy electromagnetic fields (EMF) stimulates the protein tyrosine kinases Lyn and Syk, results in tyrosine phosphorylation of multiple electrophoretically distinct substrates, and leads to downstream activation of protein kinase C (PKC). EMF exposure enhances protein tyrosine phosphorylation in Syk deficient but not in Lyn-deficient B-lineage lymphoid cells and stimulates Lyn kinase activity in wild type as well as Syk-deficient B-lineage lymphoid cells. These results indicate that activation of Lyn kinase is sufficient and mandatory for EMF-induced tyrosine phosphorylation in B-lineage lymphoid cells. The PKC activity increases later than the Lyn activity and pretreatment with the PTK inhibitors genistein or herbimycin A abrogates the EMF-induced PKC signal. Thus, stimulation of Lyn is a proximal and mandatory step in EMF-induced activation of PKC in B-lineage lymphoid cells. Our observations prompt the hypothesis that a delicate growth regulatory balance might be altered in B-lineage lymphoid cells by EMF-induced activation of Lyn. PMID- 7499234 TI - Interplay of Sp1 and Egr-1 in the proximal platelet-derived growth factor A-chain promoter in cultured vascular endothelial cells. AB - The platelet-derived growth factor (PDGF) A-chain has been implicated in the initiation and progression of vascular occlusive lesions. The elements in the human PDGF-A promoter that mediate increased expression of the gene in vascular endothelial cells have not been identified. A potent inducer of PDGF-A expression in endothelial cells is phorbol 12-myristate 13-acetate (PMA). 5'-Deletion and transfection analysis revealed that a G+C-rich region in the proximal PDGF-A promoter is required for PMA-inducible gene expression. This region bears overlapping consensus recognition sequences for Sp1 and Egr-1. PMA induces Egr-1 mRNA expression within 1 h, whereas PDGF-A transcript levels increase after 2-4 h. Constitutive levels of Sp1 are not altered over 24 h. A specific nucleoprotein complex is formed when an oligonucleotide bearing the G+C-rich element is incubated with nuclear extracts from PMA-treated cells. The temporal appearance of this complex is consistent with the transient increase in Egr-1 transcripts. Antibodies to Egr-1 completely supershift the PMA-induced complex. Interestingly, increased nuclear levels of Egr-1 attenuate the ability of Sp1 to interact with the oligonucleotide, implicating competition between Egr-1 and Sp1 for the G+C rich element. Binding studies with recombinant proteins demonstrate that Egr-1 can displace Sp1 from this region. Insertion of the G+C-rich element into a hybrid promoter-reporter construct confers PMA inducibility on the construct. Mutations that abolish Egr-1 binding also abrogate expression induced by PMA or overexpressed Egr-1. These findings demonstrate that PMA-induced Egr-1 displaces Sp1 from the G+C-rich element and activates expression driven by the PDGF-A proximal promoter in endothelial cells. The Sp1/Egr-1 displacement mechanism may be an important regulatory circuit in the control of inducible gene expression in vascular endothelial cells. PMID- 7499233 TI - GAL4/GAL80-dependent nucleosome disruption/deposition on the upstream regions of the yeast GAL1-10 and GAL80 genes. AB - Metabolic reactivation (incubating spheroplasts with galactose and casamino acids) causes disruption of nucleosomes from the upstream regions of the yeast GAL1, GAL10, and GAL80 genes. The disruption is specific. It depends on the transcription activator Gal4; it only occurs in galactose-reactivated chromatin from galactose-grown cells; it only affects upstream region, gene-proximal nucleosomes. Due to this specificity and because some of the same regions have shown induction-dependent changes by in vivo analyses, we suggest that the nucleosome-disrupted structure produced by reactivation is the authentic chromatin structure for these regions under conditions of galactose-induced GAL1 10 and GAL80 expression. It is necessary to carry out a spheroplast reactivation treatment in order to observe this disrupted structure in nuclear chromatin because nucleosomes are redeposited onto these regions during the preliminary steps of nuclear isolation (cell harvest/spheroplast preparation) probably in response to the nonphysiological conditions associated with these steps. However, during the same isolation procedures in cells lacking Gal80 protein, there is no nucleosome deposition on these regions, and the in vivo disrupted structure remains present in the nuclear chromatin. Therefore, the nucleosome deposition process that operates in wild-type cells is dependent on Gal80 protein, defining another activity of this negative regulator. PMID- 7499235 TI - In vivo assembly of the proteasomal complexes, implications for antigen processing. AB - The multicatalytic and multisubunit proteasomal complexes have been implicated in the processing of antigens to peptides presented by class I major histocompatibility complex molecules. Two structural complexes of this proteinase, 20 S and 26 S proteasomes, have been isolated from cells. By analyzing in vivo assembly of the proteasomal complexes we show that the 20 S proteasomal complexes are irreversibly assembled via 15 S assembly intermediates containing unprocessed beta-type subunits. The 20 S proteasomes further associate reversibly with proteasome activators PA28 or pre-existing ATPase complexes to form 26 S proteasomal complexes. Our findings that not all of the 20 S proteasomal complexes are assembled into 26 S proteasomal complexes within cells and that all of PA28 and ATPase complexes are associated with 20 S proteasomes strongly suggest that all proteasomal complexes coexist within cells. We further demonstrate that 26 S proteasomal complexes are predominantly present in the cytoplasm and a significant portion of the 20 S proteasomal complexes is associated with the endoplasmic reticulum membrane. Taken together, our findings suggest that depending upon their associated regulatory components, 26 S and 20 S PA28 proteasomal complexes serve different housekeeping functions within the cells, while they degrade antigens in a cooperative manner in antigen processing. PMID- 7499236 TI - Transcriptional repression by the human homeobox protein EVX1 in transfected mammalian cells. AB - The human homeobox protein EVX1 (EVX1) is thought to play an important role during embryogenesis. In this study, the effect of EVX1 on gene transcription has been investigated in transfected mammalian cells. EVX1 expression represses transcription of a reporter gene directed by either cell-specific or viral promoter/enhancer sequences in a variety of mammalian cell lines and in a concentration-dependent manner. Transcriptional repression is independent of the presence of DNA-binding sites for EVX1 in all the promoters we tested. Furthermore, repression by EVX1 is evident also using a TATA-less minimal promoter in the reporter construct. A carboxyl-terminal proline/alanine-rich region of EVX1 seems to be responsible for the transcriptional repression activity, as suggested by transfection of EVX1 mutants. We speculate that the repressor function of EVX1 contributes to its proposed role in embryogenesis. PMID- 7499237 TI - DNA structure determines protein binding and transcriptional efficiency of the proenkephalin cAMP-responsive enhancer. AB - Two precisely arranged proenkephalin cAMP response elements (CREs) behave as a single protein binding site. The experiments described support a model in which a secondary structural change creates a new binding site, which is made up of sequences from both of the CREs. The CRE-binding protein (CREB) binds CRE-1, but binding there is entirely dependent on the presence of CRE-2. Electron spectroscopic images show that a CREB dimer occupies twice as much DNA in the proenkephalin gene as in the prodynorphin gene. The enhancer region is sensitive to P1 nuclease in a CREB concentration-dependent manner, and sensitivity is strand-specific, indicating protein-stabilized structural change. DNase I analysis shows that in the native proenkephalin gene, CREB binds both CRE-1 and CRE-2. In vivo, both CREs are occupied in the transcriptionally active proenkephalin gene, while neither is in the silent gene. Whereas CREB can bind CRE-2, mutation or elimination of either proenkephalin CRE alters response to second messengers and transcription factors. Thus, binding to CRE-2 alone is not sufficient. Specific and efficient transcription of the proenkephalin gene requires the presence of both CREs, precisely arranged to allow them to form a single protein binding site. PMID- 7499238 TI - Use of a new rat chondrosarcoma cell line to delineate a 119-base pair chondrocyte-specific enhancer element and to define active promoter segments in the mouse pro-alpha 1(II) collagen gene. AB - We show that a new rat chondrosarcoma (RCS) cell line established in long-term culture from the Swarm tumor displayed a stable differentiated chondrocyte-like phenotype. Indeed, these cells produced the collagen types II, IX, and XI and alcian blue-stainable cartilage-specific proteoglycans, but no type I or type III collagen. To functionally characterize their chondrocytic nature, the cells were stably transfected with a type II collagen/beta geo chimeric gene which confers essentially perfect chondrocyte-specific expression in transgenic mice. RCS cells expressed both beta-galactosidase and G418 resistance, in comparison with similarly transfected 10T1/2 and NIH/3T3 fibroblasts which did not. These cells were then used to perform a systematic deletion analysis of the first intron of the mouse type II collagen gene (Col2a1) using transient expression experiments to determine which segments stimulated expression of a luciferase reporter gene in RCS cells but not in 10T1/2 fibroblasts. Cloning of two tandem copies of a 156 base pair (bp) intron 1 fragment (+2188 to +2343) in a construction containing a 314-bp Col2a1 promoter caused an almost 200-fold increase in promoter activity in RCS cells but no increase in 10T1/2 cells. DNase I footprint analysis over this 156-bp fragment revealed two adjacent protected regions, FP1 and FP2, located in the 3'-half of this segment, but no differences were seen with nuclear extracts of RCS cells and 10T1/2 fibroblasts. Deletion of FP2 to leave a 119-bp segment decreased enhancer activity by severalfold, but RCS cell specificity was maintained. Further deletions indicated that sequences both in the 5' part of the 119-bp fragment and in FP1 were needed simultaneously for RCS cell-specific enhancer activity. A series of deletions in the promoter region of the mouse Col2a1 gene progressively reduced activity when these promoters were tested by themselves in transient expression experiments. However, these promoter deletions were all activated to a similar level in RCS cells by a 231-bp intron 1 fragment that included the 156-bp enhancer. The RCS cell-specific activity persisted even if the Col2a1 promoter was replaced by a minimal adenovirus major late promoter. This 231-bp intron 1 fragment also had strong enhancing activity in transiently transfected mouse primary chondrocytes. Our experiments establish the usefulness of RCS cells as an experimental system for studies of the control of chondrocyte specific genes, provide an extensive delineation of segments in the Col2a1 first intron involved in chondrocyte-specific activity, and show that promoter sequences are dispensable for chondrocyte specificity. PMID- 7499239 TI - Characterization of deletion and truncation mutants of the rat glucagon receptor. Seven transmembrane segments are necessary for receptor transport to the plasma membrane and glucagon binding. AB - Glucagon receptor mutants were characterized with the aim of elucidating minimal structural requirements for proper biosynthesis, ligand binding, and adenylyl cyclase coupling. One N-terminal deletion mutant and five truncation mutants with progressively shorter C termini were expressed in transiently transfected monkey kidney (COS-1) cells. Each truncation mutant was designed so that the truncated C terminal tail would remain on the cytoplasmic surface of the receptor. In order to characterize the cellular location of the expressed receptor mutants, a highly specific, high affinity antipeptide antibody was prepared against the extracellular, N-terminal tail of the receptor. Immunoblot analysis and immunofluorescence microscopy showed that the presence of all seven putative transmembrane segments, but not not an intact N-terminal tail, was required for cell surface expression of the receptor. Membranes from cells expressing receptor mutants lacking a large portion of the N-terminal tail or any of the seven putative transmembrane segments failed to bind glucagon. Membranes from cells expressing the C-terminal tail truncation mutants, which retained all seven transmembrane segments, bound glucagon with affinities similar to that of the native receptor and activated cellular adenylyl cyclase in response to glucagon. These results indicate that all seven helices are necessary for the proper folding and processing of the glucagon receptor. Glycosylation is not required for the receptor to reach the cell surface, and it may not be required for ligand binding. However, the N-terminal extracellular portion of the receptor is required for ligand binding. Most of the distal C-terminal tail is not necessary for ligand binding, and the absence of the tail may increase slightly the receptor binding affinity for glucagon. The C-terminal tail is also not necessary for adenylyl cyclase coupling and therefore does not play a direct role in G protein (GS) activation by the glucagon receptor. PMID- 7499240 TI - Structural organization and chromosomal assignment of the human obese gene. AB - The obese (ob) gene has been identified through a positional cloning approach; the mutation of this gene causes marked hereditary obesity and diabetes mellitus in mice. We report here the isolation and characterization of the human ob gene. Southern blot analysis demonstrated a single copy of the ob gene in the human genome. The human ob gene spanned approximately 20 kilobases (kb) and contained three exons separated by two introns. The first intron, approximately 10.6 kb in size, occurred in the 5'-untranslated region, 29 base pair (bp) upstream of the ATG start codon. The second intron of 2.3 kb in size was located at glutamine +49. By rapid amplification of 5'-cDNA ends, the transcription initiation sites were mapped 54-57 bp upstream of the ATG start codon. The 172-bp 5'-flanking region of the human ob gene contained a TATA box-like sequence and several cis acting regulatory elements (three copies of GC boxes, an AP-2-binding site, and a CCAAT/enhancer-binding protein-binding site). By the fluorescence in situ hybridization technique, the ob gene was assigned to human chromosome 7q31.3. This study should establish the genetic basis for ob gene research in humans, thereby leading to the better understanding of the molecular mechanisms underlying the ob gene. PMID- 7499241 TI - Increased expression of CD44 in bovine articular chondrocytes by catabolic cellular mediators. AB - Bovine articular chondrocytes cultured in alginate beads were used to study the effect of catabolic cellular mediators on CD44 expression. Treatment with either the 29-kDa fragment of fibronectin or interleukin-1 alpha results in a time- and dose-dependent inhibition of proteoglycan synthesis as well as a stimulation in the expression of CD44 mRNA level as determined by semi-quantitative polymerase chain reaction following reverse transcription. No noticeable effect at 6 h was observed. By 24 h, the major CD44 product (CD44H) from fibronectin fragment treated cultures showed an 8-fold increase; CD44H from interleukin-1 alpha treated cultures showed a 6-fold increase as compared to control cultures. In addition, a minor band, determined to be an isoform of CD44, was also shown to be up-regulated by both mediators. Stimulation of CD44 mRNA via interleukin-1 was also evident by in situ hybridization studies of bovine as well as human articular cartilage in organ culture. The increased in CD44 mRNA is matched by an increase at the protein level as determined by Western blot analysis. The Western blot reveals a doublet protein band at 80-90 kDa that corresponds to the molecular mass of CD44H. Cultures incubated with fibronectin fragments for 24 h had an 8.0-fold increase in CD44, while a 6.6-fold was observed for interleukin-1 alpha. Fluorescein-conjugated hyaluronan binding and internalization studies indicate that the increase in CD44 protein, induced by interleukin-1 alpha, closely correlates with an increase in functional hyaluronan receptors present at the chondrocyte cell surface. Taken together these results indicate that conditions that up-regulate chondrocyte catabolism also up-regulate the expression of CD44, a cell surface hyaluronan receptor involved in hyaluronan endocytosis. PMID- 7499242 TI - Identification and characterization of a novel related adhesion focal tyrosine kinase (RAFTK) from megakaryocytes and brain. AB - We have isolated a cDNA encoding a novel human intracytoplasmic tyrosine kinase, termed RAFTK (for a related adhesion focal tyrosine kinase). In addition, we have cloned and characterized the murine homolog of the human RAFTK cDNA. Comparison of the deduced amino acid sequences of human RAFTK and murine Raftk cDNAs revealed 95% homology, indicating that RAFTK is highly conserved between these species. The RAFTK cDNA clone, encoding a polypeptide of 1009 amino acids, has closest homology (48% identity, 65% similarity) to the focal adhesion kinase (pp125FAK). Comparison of the deduced amino acid sequences also indicates that RAFTK, like pp125FAK, lacks a transmembrane region, myristylation sites, and SH2 and SH3 domains. In addition, like pp125FAK, RAFTK contains a kinase domain flanked by large N-terminal (426 residues) and C-terminal (331 residues) domains, and the C-terminal region contains a predicted proline-rich stretch of residues. In fetal tissues, RAFTK expression was abundant in brain, and low levels were observed in lung and liver. In adult tissues, it was less restricted, indicating that RAFTK expression is developmentally up-regulated. Expression of RAFTK was also observed in human CD34+ marrow cells, primary bone marrow megakaryocytes, platelets, and various areas of brain. The human RAFTK gene was assigned to human chromosome 8 using genomic DNAs from human/rodent somatic cell hybrid lines. The mouse Raftk gene was mapped to chromosome 14, closely linked to gonadotropin releasing hormone. Using specific antibodies for RAFTK, a approximately 123-kDa protein from the human megakaryocytic CMK cell line was immunoprecipitated. Treatment of the megakaryocytic CMK cells with thrombin caused a rapid induction of tyrosine phosphorylation of RAFTK protein. The structural features of RAFTK suggest that it is a member of the focal adhesion kinase gene family and may participate in signal transduction in human megakaryocytes and brain as well as in other cell types. PMID- 7499244 TI - Differential sensitivity of interleukin-1 alpha and -beta precursor proteins to cleavage by calpain, a calcium-dependent protease. AB - In view of the observations that the calcium ionophores, A23187 and ionomycin, enhance the processing and secretion of interleukin-1 (IL-1 alpha) and IL-1 beta from macrophages, and IL-1 alpha processing is mediated by calpain, a calcium dependent protease, we evaluated the possibility that calpain might also play a role in the processing of IL-1 beta. Whereas calpain-containing P388D1 macrophage lysates and purified calpain processed precursor IL-1 alpha to its mature 17-kDa form, precursor IL-1 beta was degraded by both sources of calpain. However, the activation of calpain in P388D1 cells that were transiently transfected with a cDNA expression vector encoding the precursor form of IL-1 beta did not result in the degradation of precursor IL-1 beta, but did result in the processing and secretion of IL-1 alpha, implying that precursor IL-1 beta is protected from calpain degradation in vivo. Furthermore, calpain did not enhance the processing of the IL-1 beta precursor by the IL-1 beta-converting enzyme. These results indicate that calpain is not involved in the processing of precursor IL-1 beta in vitro or in vivo. The IL-1 beta precursor may be protected from calpain degradation by a sequestering mechanism that involves a cytoplasmic factor(s) that reduces the sensitivity of IL-1 beta to attack by calpain or localizes IL-1 beta to a site that precludes any interaction with the protease. Although MDL 28,170, a calpain inhibitor, prevented the ionomycin-induced processing of precursor IL-1 alpha to the mature protein in P388D1 cells, it did not inhibit the ionomycin-induced secretion of the mature IL-1 alpha and -beta proteins expressed in these cells. These results indicate that a calcium-dependent factor other than calpain is involved in the secretion of the mature IL-1 proteins. PMID- 7499243 TI - Non-neuronal enolase is an endothelial hypoxic stress protein. AB - The hypoxia-associated proteins (HAPs) are five cell-associated stress proteins (M(r) 34, 36, 39, 47, and 57) up-regulated in cultured vascular endothelial cells (EC) exposed to hypoxia. While hypoxic exposure of other cell types induces heat shock and glucose-regulated proteins, EC preferentially up-regulate HAPs. In order to identify the 47-kDa HAP, protein from hypoxic bovine EC lysates was isolated, digested with trypsin, and sequenced. Significant identity was found with enolase, a glycolytic enzyme. Western analyses confirmed that non-neuronal enolase (NNE) is up-regulated in hypoxic EC. Western analysis of subcellular fractions localized NNE primarily to the cytoplasm and confirmed that it was up regulated 2.3-fold by hypoxia. Interestingly, NNE also appeared in the nuclear fraction of EC but was unchanged by hypoxia. Northern analyses revealed that NNE mRNA hypoxic up-regulation began at 1-2 h, peaked at 18 h, persisted for 48 h, and returned to base line after return to 21% O2 for 24 h. Hypoxia maximally up regulated NNE mRNA levels 3.4-fold. While hypoxic up-regulation of NNE may have a protective effect by augmenting anaerobic metabolism, we speculate that enolase may contribute to EC hypoxia tolerance through one or more of its nonglycolytic functions. PMID- 7499245 TI - Characterization of lysosomal acid lipase by site-directed mutagenesis and heterologous expression. AB - Lysosomal acid lipase (LAL) is essential for the hydrolysis of cholesterol esters and triglycerides that are delivered to the lysosomes via the low density lipoprotein receptor system. The deficiency of LAL is associated with cholesteryl ester storage disease (CESD) and Wolman's disease (WD). We cloned the human LAL cDNA and expressed the active enzyme in the baculovirus system. Two molecular forms (M(r) approximately 41,000 and approximately 46,000) with different glycosylation were found intracellularly, and approximately 24% of the M(r) approximately 46,000 form was secreted into the medium. Tunicamycin treatment produced only an inactive M(r) approximately 41,000 form. This result implicates glycosylation occupancy in the proper folding for active-site function. Catalytic activity was greater toward cis- than trans-unsaturated fatty acid esters of 4 methylumbelliferone and toward esters with 7-carbon length acyl chains. LAL cleaved cholesterol esters and mono-, tri-, and diglycerides. Heparin had a biphasic effect on enzymatic activity with initial activation followed by inhibition. Inhibition of LAL activity by tetrahydrolipstatin and diethyl p nitrophenyl phosphate suggested the presence of active serines in binding/catalytic domain(s) of the protein. Site-directed mutagenesis at two putative active centers, GXSXG, showed that Ser153 was important to catalytic activity, whereas Ser99 was not and neither was the catalytic nucleophile. Three reported mutations (L179P, L336P, and delta AG302 deletion) from CESD patients were created and expressed in the Sf9 cell system. None cleaved cholesterol esters, and L179P and L336P cleaved only triolein at approximately 4% of wild type levels. These results suggest that mechanisms, in addition to LAL defects, may operate in the selective accumulation of cholesterol esters or triglycerides in CESD and WD patients. PMID- 7499246 TI - A consensus insulin response element is activated by an Ets-related transcription factor. AB - Insulin increases expression of somatostatin-chloramphenicol acetyltransferase (CAT) constructs 10-fold and thymidine kinase-CAT constructs 5-fold in GH4 cells. These responses are similar to our previously reported data on insulin-increased prolactin-CAT expression. They are also observed in HeLa cells and are thus not cell type specific. The evidence suggests that the insulin responsiveness of these genes is mediated by an Ets-related transcription factor. First, linker scanning mutations and/or deletions of the prolactin, somatostatin, and thymidine kinase promoters suggest that their insulin responsiveness is mediated by the sequence CGGA. This sequence is identical with the response element of the Ets related transcription factors. Second, CGGA-containing sequences placed at -88 in the delta MTV-CAT reporter plasmid conferred insulin responsiveness to the mammary tumor virus promoter. Third, expression of the DNA-binding domain of c Ets-2, which acts by blocking effects mediated by Ets-related transcription factors, inhibits the response of these promoters to insulin. Finally, the Ets related proteins Sap and Elk-1 bind to the prolactin, somatostatin, and thymidine kinase insulin-response elements. An Ets-like element was found in all insulin sensitive promoters examined and may serve a similar function in those promoters. PMID- 7499247 TI - Hepatocyte growth factor-induced scatter of Madin-Darby canine kidney cells requires phosphatidylinositol 3-kinase. AB - Hepatocyte growth factor/scatter factor (HGF/SF) is a multifunctional cytokine that induces mitogenesis, motility, invasion, and morphogenesis of several epithelial and endothelial cell lines in culture. The receptor for HGF/SF has been identified as the Met tyrosine kinase. To investigate the signaling pathways that are involved in these events, we have generated chimeric receptors containing the extracellular domain of the colony-stimulating factor-1 (CSF-1) receptor fused to the transmembrane and intracellular domains of the Met receptor (MET). Madin-Darby canine kidney (MDCK) epithelial cells expressing the CSF-MET chimera dissociate and scatter in response to CSF-1. However, cells expressing a mutant CSF-MET receptor containing a phenylalanine substitution for tyrosine 1356 were unable to scatter or form branching tubules following stimulation with CSF 1. Tyrosine 1356 is essential for the recruitment of multiple substrates including the p85 subunit of PI3-kinase, phospholipase C gamma, and Grb2. In this study, we have investigated the role of PI3-kinase and a downstream target of PI3 kinase, pp70S6K, in the induction of MDCK cell scatter in response to HGF/SF. Our results demonstrate that following stimulation with HGF/SF, activation of PI3 kinase but not pp70S6K is essential for MDCK cell scatter. PMID- 7499249 TI - Maturation of pre-apocytochrome f in vivo. A site-directed mutagenesis study in Chlamydomonas reinhardtii. AB - The biosynthesis of cytochrome f is a multistep process which requires processing of the precursor protein and covalent ligation of a c-heme upon membrane insertion of the protein. The crystal structure of a soluble form of cytochrome f has revealed that one axial ligand of the c-heme is provided by the alpha-amino group of Tyr1 generated upon cleavage of the signal sequence from the precursor protein (Martinez S. E., Huang D., Szczepaniak A., Cramer W.A., and Smith J. L. (1994) Structure 2, 95-105). We therefore investigated, by site-directed mutagenesis, the possible interplay between protein processing and heme attachment to cytochrome f in Chlamydomonas reinhardtii. These modifications were performed by chloroplast transformation using a petA gene encoding the full length precursor protein and also a truncated version lacking the C-terminal membrane anchor. We first substituted the two cysteinyl residues responsible for covalent ligation of the c-heme, by a valine and a leucine, and showed that heme binding is not a prerequisite for cytochrome f processing. In another series of experiments, we replaced the consensus cleavage site for the thylakoid processing peptidase, AQA, by an LQL sequence. The resulting transformants were nonphototrophic and displayed delayed processing of the precursor form of cytochrome f, but nonetheless both the precursor and processed forms showed heme binding and assembled in cytochrome b6f complexes. Thus, pre-apocytochrome f adopts a suitable conformation for the cysteinyl residues to be substrates of the heme lyase and pre-holocytochrome f folds in an assembly-competent conformation. In the last series of experiments, we compared the rates of synthesis and degradation of the various forms of cytochrome f in the four types of transformants under study: (i) the C terminus membrane anchor apparently down regulates the rate of synthesis of cytochrome f and (ii) degradation of misfolded forms of cytochrome f occurs by a proteolytic system intimately associated with the thylakoid membranes. PMID- 7499250 TI - Characterization of S-AKAP84, a novel developmentally regulated A kinase anchor protein of male germ cells. AB - In mammalian spermatozoa, most of the type II alpha isoform of cAMP-dependent protein kinase (PKAII alpha) is anchored at the cytoplasmic surface of a specialized array of mitochondria in the flagellar cytoskeleton. This places the catalytic subunits of PKAII alpha in proximity with potential target substrates in the cytoskeleton. The mechanism by which PKAII alpha is anchored at the outer surface of germ cell mitochondria has not been elucidated. We now report the cloning of a cDNA that encodes a novel, germ cell A kinase anchor protein (AKAP) designated S-AKAP84. S-AKAP84 comprises 593 amino acids and contains a centrally located domain that avidly binds regulatory subunits (RII alpha and RII beta) of PKAII alpha and PKAII beta. The 3.2-kilobase S-AKAP84 mRNA and the cognate S AKAP84 RII binding protein are expressed principally in the male germ cell lineage. Expression of S-AKAP84 is tightly regulated during development. The protein accumulates as spermatids undergo nuclear condensation and tail elongation. The timing of S-AKAP84 expression is correlated with the de novo accumulation of RII alpha and RII beta subunits and the migration of mitochondria from the cytoplasm (round spermatids) to the cytoskeleton (midpiece in elongating spermatids). Residues 1-30 at the NH2 terminus of S-AKAP84 constitute a putative signal/anchor sequence that may target the protein to the outer mitochondrial membrane. Immunofluorescence analysis demonstrated that S-AKAP84 is co-localized with mitochondria in the flagellum. PMID- 7499248 TI - GH3 cell-specific expression of Kv1.5 gene. Regulation by a silencer containing a dinucleotide repetitive element. AB - A silencer element (Kv1.5 repressor element; KRE) was characterized by deletion analyses in the promoter of Kv1.5, a voltage-gated potassium channel. The silencer element selectively decreases expression of Kv1.5- and thymidine kinase chloramphenicol acetyl-transferase reporter gene constructs in cell lines that do not express Kv1.5 polypeptide. It contains a dinucleotide repetitive element (poly(GT)19(GA)1(CA)15(GA)16), and self-associates spontaneously in vitro to form complexes with slow electrophoretic mobility. Deletion of the repetitive element abolished self-association in vitro and the silencing activity in transient transfection experiments in vivo. Electromobility gel shift assays of KRE with GH3 cells nuclear extracts detected the formation of a unique DNA-protein complex, which was not detectable in Chinese hamster ovary and COS-7 cells. This complex does not react with an antibody against nonhistone high mobility group 1 protein, which binds KRE in gel retardation assays. These observations establish that a dinucleotide tandem repeat sequence, capable of self-association, forms part of a cell-specific silencer element in a mammalian gene. PMID- 7499251 TI - Two residues that may ligate Ca2+ in transmembrane domain six of the plasma membrane Ca(2+)-ATPase. AB - In order to identify Ca2+ ligands in the putative transmembrane domain 6 of the plasma membrane Ca2+ pump, amino acids Asn879, Met882, Asp883, and Ser887 were singly altered. Asn879, Met882, and Asp883 were chosen because the corresponding amino acids have been proposed as Ca2+ ligands in the sarcoplasmic reticulum Ca2+ pump (Clarke, D. M., Loo, T. W., and MacLennan, D. H. (1990) J. Biol. Chem. 265, 6262-6267). For the alterations, a fully active truncated version of the pump was used, because the interaction of Ca2+ with the pump could be studied without interference from calmodulin binding. The mutants at Asn and Asp did not carry out ATP-supported Ca2+ uptake and formed no acylphosphate from [gamma-32P]ATP, suggesting that, like the corresponding amino acids in the sarcoplasmic reticulum Ca2+ pump, these two are Ca2+ ligands. However, all the mutants at the position of Met882 showed some activity. Indeed, the Met882--> Ile mutant was fully active at a saturating Ca2+ concentration and only the K1/2 for Ca2+ activation was shifted slightly upward. Converting the Met to Thr (which is the corresponding residue in the sarcoplasmic reticulum Ca2+ pump) reduced the activity to 20% of the wild type, further emphasizing the differences between the two Ca2+ pumps. The mutant Ser887--> Ala was expressed in greater amounts than, and had a specific activity about 50% higher than, the wild type, indicating that this serine also could not be a Ca2+ ligand and could not replace the missing Thr at position Met882. PMID- 7499252 TI - Identification of a new adapter protein that may link the common beta subunit of the receptor for granulocyte/macrophage colony-stimulating factor, interleukin (IL)-3, and IL-5 to phosphatidylinositol 3-kinase. AB - Binding of human granulocyte/macrophage colony-stimulating factor (hGM-CSF) to its receptor induces the rapid activation of phosphatidylinositol-3 kinase (PI 3 kinase). As hGM-CSF receptor (hGMR) does not contain a consensus sequence for binding of PI 3-kinase, hGMR must use a distinct mechanism for its association with and activation of PI 3-kinase. Here, we describe the identification of a tyrosine-phosphorylated protein of 76-85 kDa (p80) that associates with the common beta subunit of hGMR and with the SH2 domains of the p85 subunit of PI 3 kinase in hGM-CSF-stimulated cells. Src/Yes and Lyn were tightly associated with the p80.PI 3-kinase complex, suggesting that p80 and other phosphotyrosyl proteins present in the complex were phosphorylated by Src family kinases. Tyrosine phosphorylation of p80 was only detected in hGM-CSF or human interleukin 3-stimulated cells, suggesting that activation of p80 might be specific for signaling via the common beta subunit. We postulate that p80 functions as an adapter protein that may participate in linking the hGM-CSF receptor to the PI 3 kinase signaling pathway. PMID- 7499253 TI - Oct-1 protein promotes functional transcription complex assembly on the mouse mammary tumor virus promoter. AB - The ubiquitous transcription factor Oct-1 stimulates basal transcription from the mouse mammary tumor virus (MMTV) promoter by binding to octamer-related sequences present in the proviral long terminal repeat. The mechanism of transcriptional activation by Oct-1 was investigated using in vitro transcription assays with a HeLa cell nuclear extract depleted of endogenous Oct-1. Oct-1-mediated transcriptional activation could be reconstituted by addition of bacterially expressed recombinant Oct-1 protein. The stimulatory effect of Oct-1 was observed only when the protein was present during formation of transcription preinitiation complexes and not when added to fully assembled complexes. Furthermore, assembled MMTV preinitiation complexes were resistant to inhibition by a competitor oligonucleotide containing MMTV octamer-related elements that could eliminate Oct 1-mediated stimulation when present during the assembly process. The time course of transcription complex assembly revealed that Oct-1 increases the number of templates on which functional transcription complexes form. Finally, experiments designed to exploit the sensitivity of discrete steps in transcription complex assembly to the anionic detergent Sarkosyl demonstrated that Oct-1 must be present during formation of an early intermediate in the assembly process. PMID- 7499254 TI - Cross-desensitization of chemoattractant receptors occurs at multiple levels. Evidence for a role for inhibition of phospholipase C activity. AB - To define the molecular mechanisms of cross-regulation among chemoattractant receptors, we stably coexpressed, in a rat basophilic leukemia (RBL-2H3) cell line, epitope-tagged receptors for the chemoattractants formylmethionylleucylphenylalanine (fMLP), a peptide of the fifth component of the complement system (C5a), and interleukin-8 (IL-8). All the expressed receptors underwent homologous phosphorylation and desensitization upon agonist stimulation. When co-expressed, epitope-tagged C5a receptor (ET-C5aR) and epitope tagged IL-8 receptor (ET-IL-8RA) were cross-phosphorylated by activation of the other. Activation of epitope-tagged fMLP receptor (ET-FR) also cross phosphorylated ET-C5aR and ET-IL-8RA, but ET-FR was totally resistant to cross phosphorylation. Similarly, C5a and IL-8 stimulation of [35S]guanosine 5'-3-O (thio) triphosphate (GTP gamma S) binding and Ca2+ mobilization were cross desensitized by each other and by fMLP. Stimulation of [35S]GTP gamma S binding by fMLP was also not cross-desensitized by C5a or IL-8, however, Ca2+ mobilization was, suggesting a site of inhibition distal to G protein activation. Consistent with this desensitization of Ca2+ mobilization, inositol 1,4,5 trisphosphate release in RBL-2H3 cells expressing both ET-C5aR and ET-FR revealed that fMLP and C5a cross-desensitized each other's ability to stimulate phosphoinositide hydrolysis. Taken together, these results indicate that receptor cross-phosphorylation correlates directly with desensitization at the level of G protein activation. The ET-FR was resistant to this process. Of note, cross desensitization of ET-FR at the level of phosphoinositide hydrolysis and Ca2+ mobilization was demonstrated in the absence of receptor phosphorylation. This suggests a new form of chemoattractant cross-regulation at a site distal to receptor/G protein coupling, involving the activity of phospholipase C. PMID- 7499256 TI - Ligand binding characteristics of the carboxyl-terminal domain of the cytokine receptor homologous region of the granulocyte colony-stimulating factor receptor. AB - The carboxyl-terminal domain (BC domain, roughly 100 amino acid residues) of the cytokine receptor homologous region in the receptor for murine granulocyte colony stimulating factor was secreted as a maltose binding protein fusion into the Escherichia coli periplasm. The murine BC domain was prepared from the fusion protein by restriction protease factor Xa digestion and was purified to homogeneity. The purified BC domain specifically and stoichiometrically bound granulocyte colony-stimulating factor. This result indicates that the BC domain is also critical for ligand binding, as shown for the amino-terminal domain of the cytokine receptor homologous region (Hiraoka, O., Anaguchi, H., Yamasaki, K., Fukunaga, R., Nagata, S., and Ota, Y. (1994) J. Biol. Chem. 269, 22412-22419). The tertiary folding and the beta-sheet structure of the BC domain were confirmed by NMR spectroscopy. The disulfide bond pattern suggested from peptide mapping was Cys224-Cys271 and Cys242-Cys285. Disruption of the disulfide bonds suggested that both bonds are critical for maintaining the folding of the BC domain, although a BC domain lacking the second bond still retained ligand binding activity. Mutational analysis of the WSXWS sequence conserved in the cytokine receptor family suggested that this motif is critical for protein folding rather than for ligand binding. PMID- 7499255 TI - Immunoadsorption of hepatic vesicles carrying newly synthesized dipeptidyl peptidase IV and polymeric IgA receptor. AB - Hepatocytes must transport newly synthesized apical membrane proteins from the basolateral to the apical plasma membrane. Our earlier morphological study showed that the apical proteins share a late (subapical) part of the transcytotic pathway with the well characterized polymeric immunoglobulin A receptor (Barr, V. A., and Hubbard, A. L. (1993) Gastroenterology 105, 554-571). Starting with crude microsomes from the livers of [35S]methionine-labeled rats, we sequentially immunoadsorbed first vesicles containing the endocytic asialoglycoprotein receptor and then (from the depleted supernatant) vesicles containing the polymeric IgA receptor. Biochemical characterization indicated that early basolateral and late endosomes were present in the first population but not in the second. Neither Golgi-, apical plasma membrane (PM)-, nor basolateral PM derived vesicles were significant contaminants of either population. Both vesicle populations contained 35S-labeled receptor and 35S-labeled-dipeptidyl peptidase IV. Importantly, the elevated relative specific activity of the dipeptidyl peptidase (% of 35S-labeled/% immunoblotted) in the second population indicated that these vesicles must transport newly synthesized dipeptidyl peptidase IV. A distinct kind of vesicle was immunoadsorbed from a "carrier-vesicle fraction"; surprisingly, these vesicles contained little 35S-receptor and virtually no dipeptidyl peptidase IV. These results, together with previous kinetic data from in vivo experiments, are consistent with a computer-generated model predicting that newly synthesized dipeptidyl peptidase IV is delivered to basolateral endosomes, which also contain newly synthesized polymeric immunoglobulin A receptor. The two proteins are then transcytosed together to the subapical region. PMID- 7499257 TI - Effects of protein S and factor Xa on peptide bond cleavages during inactivation of factor Va and factor VaR506Q by activated protein C. AB - Inactivation of membrane-bound factor Va by activated protein C (APC) proceeds via a biphasic reaction that consists of a rapid and a slow phase, which are associated with cleavages at Arg506 and Arg306 of the heavy chain of factor Va, respectively. We have investigated the effects of protein S and factor Xa on APC catalyzed factor Va inactivation. Protein S accelerates factor Va inactivation by selectively promoting the slow cleavage at Arg306 (20-fold). Factor Xa protects factor Va from inactivation by APC by selectively blocking cleavage at Arg506. Inactivation of factor VaR506Q, which was isolated from the plasma of a homozygous APC-resistant patient and which lacks the Arg506 cleavage site, was also stimulated by protein S but was not affected by factor Xa. This confirms that the target sites of protein S and factor Xa involve Arg306 and Arg506, respectively. Factor Xa completely blocked APC-catalyzed cleavage at Arg506 in normal factor Va (1 nM) with a half-maximal effect (K1/2Xa) at 1.9 nM factor Xa. Expression of cofactor activity of factor Va in prothrombin activation required much lower factor Xa concentrations (K1/2Xa = 0.08 nM). When the ability of factor Xa to protect factor Va from inactivation by APC was determined at low factor Va concentrations during prothrombin activation much lower amounts of factor Xa were required (K1/2Xa = 0.03 nM). This indicates 1) that factor Va is optimally protected from inactivation by APC by incorporation into the prothrombinase complex during ongoing prothrombin activation, and 2) that the formation of a catalytically active prothrombinase complex and protection of factor Va from inactivation by APC likely involves the same interaction of factor Xa with factor Va. In accordance with the proposed mechanisms of action of protein S and factor Xa, we observed that the large differences between the rates of APC-catalyzed inactivation of normal factor Va and factor VaR506Q were almost annihilated in the presence of factor Xa and protein S. This observation may explain why, in the absence of other risk factors, APC resistance only results in a weak prothrombotic condition. PMID- 7499258 TI - Cloning and functional analysis of the ndk1 gene encoding nucleoside-diphosphate kinase in Schizosaccharomyces pombe. AB - We cloned the ndk1 gene encoding a subunit of nucleoside-diphosphate kinase (NDK) from Schizosaccharomyces pombe, by using polymerase chain reaction. The deduced ndk1 gene product has 151 amino acid residues and is approximately 60% identical with both Saccharomyces cerevisiae and mammalian NDKs. The gene product exhibited NDK activity and cross-reacted with antibodies raised against rat NDK. Disruption of ndk1 greatly reduced the cellular NDK activity but caused no obvious phenotype in cell growth and sexual development of the organism. However, a mutated allele of ndk1 could inhibit sexual development in a dominant-negative manner. This allele carried a point mutation in cysteine 116, which locates next to the putative active center histidine 117, and the mutant gene product showed no NDK activity. Gene expression inducible in response to mating pheromone signaling was decreased in cells carrying the dominant-negative allele. Cases have been reported in higher eukaryotes in which NDK appears to play a more sophisticated role than a simple catalyst in cell physiology, and the results of this study suggest that S. pombe NDK may also perform such a role in regulation of sexual development in the fission yeast. PMID- 7499259 TI - Hypoxia, a novel inducer of acute phase gene expression in a human hepatoma cell line. AB - Transcriptional regulation of gene expression by hypoxia is an important, but yet only marginally characterized mechanism by which organisms adapt to low oxygen concentrations. The human hepatoma cell line HepG2 is a widely used model for studying hypoxic induction of the hematopoietic growth factor erythropoietin. In an attempt to identify additional genes expressed in HepG2 cells during hypoxia, we differentially screened a cDNA library derived from hypoxic (1% O2) HepG2 cells using probes isolated from either normoxic (21% O2) or hypoxic cells. Two genes were identified, one encoding aldolase, a member of the glycolytic enzymes, and the other encoding alpha 1-antitrypsin which belongs to the family of the acute phase (AP) responsive proteins. Whereas hypoxic induction of glycolytic enzymes is well established, oxygen-dependent regulation of AP genes has not been reported so far. AP proteins are liver-derived plasma proteins whose production during inflammation is either up-regulated (positive AP reactants) or down regulated (negative AP reactants). In the present study, we demonstrate that on the mRNA level hypoxic stimulation of HepG2 cells led to (i) an induction of the positive AP reactants alpha 1-antitrypsin, alpha 1-antichymotrypsin, complement C3, haptoglobin, and alpha 1-acid glycoprotein; (ii) a down-regulation of the negative AP reactant albumin; (iii) an up-regulation of the negative AP reactant transferrin; and (iv) unchanged levels of the positive AP reactants alpha- and beta-fibrinogen as well as hemopexin. Cycloheximide inhibited hypoxic up regulation of AP mRNAs demonstrating that de novo protein synthesis is required for hypoxic induction. Nuclear run-on assays indicate that the hypoxic increase in AP mRNAs is mainly due to transcriptional regulation. The hypoxic response was compared to AP stimulation by interleukin 6. The results suggest that the adaptive response to hypoxia overlaps with, but is not identical with, the AP response mediated by interleukin 6. PMID- 7499261 TI - Systematic introduction of proline in a eukaryotic signal sequence suggests asymmetry within the hydrophobic core. AB - The hydrophobic core or h region of both prokaryotic and eukaryotic signal sequences is the predominant structural domain that controls the efficiency of protein translocation across membranes. Characteristically, hydrophobic cores appear to assume alpha-helical conformations, and studies in prokaryotes have indicated that this conformation is necessary for efficient signal sequence function. To address the conformational constraints of a eukaryotic signal sequence, we have introduced a single proline in almost each position of the signal sequence hydrophobic core of glycoprotein C (gC) of the swine herpesvirus, pseudorabies virus. When the resulting mutant virus strains were used to infect cells, we found that substitution of proline at certain positions affected gC translocation greater than its introduction at other sites within the hydrophobic core. The observed positional effects did not completely correlate with reductions in overall hydrophobicity or linear position within the hydrophobic core. Rather, it appeared that one face of the gC signal sequence alpha-helix is far more sensitive to proline disruption than the other, potentially indicating a functional asymmetry. PMID- 7499262 TI - Ras is not required for the interleukin 3-induced proliferation of a mouse pro-B cell line, BaF3. AB - It has been demonstrated that Ras is involved in interleukin 3 (IL-3)-stimulated signal transduction in various hematopoietic cultured cells (Satoh, T., Nakafuku, M., Miyajima, A., and Kaziro, Y. (1991) Proc. Natl. Acad. Sci. U.S.A. 88, 3314 3318; Duronio, V., Welham, M. J., Abraham, S., Dryden, P., and Schrader, J. W. (1992) Proc. Natl. Acad. Sci. U.S.A. 89, 1587-1591). However, it has not been fully understood which of IL-3-promoted cellular responses, i.e. proliferation, survival, and differentiation, requires Ras function. We employed a system of inducible expression of the dominant-negative (S17N) or dominant-active (G12V) mutant of Ras in BaF3 mouse pro-B cell line to analyze the role of Ras in IL-3 stimulated signal transduction. Induction of the dominant-negative Ras(S17N) effectively inhibited the IL-3-induced activation of c-Raf-1 and mitogen activated protein kinase (MAPK). Furthermore, the activation of fos gene promoter following IL-3 stimulation was almost completely abolished when Ras(S17N) was induced. Under these conditions, Ras(S17N) exhibited no inhibitory effect on IL-3 dependent proliferation assessed by the increase of cell numbers and a mitochondrial enzyme activity. The results indicate that Ras-dependent pathways, including the Raf/MAPK/Fos pathway, are dispensable for IL-3-induced growth stimulation. When BaF3 cells were treated with a tyrosine kinase inhibitor, herbimycin A, IL-3-dependent proliferation of the cells was impaired, suggesting that tyrosine kinase-mediated pathways are critical for growth promotion. On the other hand, apoptotic cell death caused by deprivation of IL-3 was prevented by the induction of the activated mutant Ras(G12V), although the rate of cell number increase was markedly reduced. Thus, it is likely that Ras-independent pathways play important roles to facilitate the proliferation although they may not be essential for IL-3-stimulated antiapoptotic signal transduction. PMID- 7499263 TI - Volume-sensitive chloride channel activity does not depend on endogenous P glycoprotein. AB - To determine whether endogenous P-glycoprotein, the MDR1 gene product that functions as a drug transport pump, is a volume-sensitive Cl- channel molecule or a protein kinase C-mediated regulator of the Cl- channel, whole-cell patch-clamp and molecular biological experiments were carried out in a human small intestinal epithelial cell line. Endogenous expression of P-glycoprotein was confirmed by Northern blot analysis, reverse transcription-polymerase chain reaction, Western blot analysis, and immunostaining. The P-glycoprotein expression was abolished by the antisense (but not sense) oligonucleotide for the MDR1 gene, whereas the magnitude of the Cl- current activated by osmotic swelling was not distinguishable between both antisense- and sense-treated cells. The volume sensitive Cl- currents were not specifically affected by the anti-P-glycoprotein monoclonal antibodies, MRK16, C219, and UIC2. An inhibitor of P-glycoprotein mediated pump activity, verapamil, was found to never affect the Cl- current. A substrate for the P-glycoprotein-mediated drug pump, vincristine or daunomycin, did not prevent swelling-induced activation of the Cl- current. Furthermore, the Cl- current was not affected by an activator of protein kinase C (12-O tetradecanoylphorbol-13-acetate or 1-oleoyl-2-acetyl-sn-glycerol). Thus, it is concluded that the endogenous P-glycoprotein molecule is not itself a volume sensitive Cl- channel nor a protein kinase C-mediated regulator of the channel in the human epithelial cells. PMID- 7499264 TI - Plasminogen activator inhibitor type 2 inhibits tumor necrosis factor alpha induced apoptosis. Evidence for an alternate biological function. AB - Plasminogen activator inhibitor type 2 (PAI-2) is a serine proteinase inhibitor or serpin that is a major product of macrophages in response to endotoxin and inflammatory cytokines. We have explored the role of PAI-2 in apoptotic cell death initiated by tumor necrosis factor alpha (TNF). HeLa cells stably transfected with PAI-2 cDNA were protected from TNF-induced apoptosis, whereas cells transfected with antisense PAI-2 cDNA, a control gene, or the plasmid vector alone remained susceptible. The level of PAI-2 expressed by different HeLa cell clones was inversely correlated with their sensitivity to TNF. Loss of TNF sensitivity was not a result of loss of TNF receptor binding. In contrast, PAI-2 expression did not confer protection against apoptosis induced by ultraviolet or ionizing radiation. The serine proteinase urokinase-type plasminogen activator was not demonstrated to be the target of PAI-2 action. The P1-Arg amino acid residue of PAI-2 was determined to be required for protection, because cells expressing PAI-2 with an Ala in this position were not protected from TNF mediated cell death. The results suggest that intracellular PAI-2 might be an important factor in regulating cell death in TNF-mediated inflammatory processes through inhibition of a proteinase involved in TNF-induced apoptosis. PMID- 7499260 TI - Thrombin stimulates phosphorylation of insulin-like growth factor-1 receptor, insulin receptor substrate-1, and phospholipase C-gamma 1 in rat aortic smooth muscle cells. AB - It has recently been reported that protein-tyrosine kinase activity is required for thrombin-induced growth in vascular smooth muscle cells (VSMC). In the present study, we have identified several phosphoproteins that are tyrosine phosphorylated in response to thrombin in quiescent VSMC. These proteins are insulin-like growth factor-1 receptor beta-subunit (IGF-IR beta), insulin receptor substrate-1 (IRS-1), and phospholipase C-gamma 1 (PLC-gamma 1). Thrombin stimulated phosphorylation of these proteins was rapid; it was maximal at 1 min and reduced thereafter. Thrombin also activated mitogen-activated protein kinases (MAPK) in quiescent VSMC in a biphasic manner with a rapid and larger peak at 10 min (6-fold) followed by a sustained smaller second peak at 2 h (2-fold). Inhibition of protein-tyrosine kinase activity by the use of two structurally different protein-tyrosine kinase inhibitors, genistein and herbimycin A, significantly blocked the thrombin-induced tyrosine phosphorylation of IGF-1R beta, IRS-1, and PLC-gamma 1 and decreased thrombin-stimulated DNA synthesis. In contrast, however, inhibition of protein-tyrosine kinase activity had no effect on thrombin activation of MAPK. Collectively, these findings suggest a role for tyrosine phosphorylation of IGF-IR beta, IRS-1, and PLC-gamma 1 in thrombin induced mitogenic signaling events in VSMC. Furthermore, while protein tyrosine phosphorylation is essential for thrombin-induced DNA synthesis, it is not required for thrombin-stimulated MAPK activation. Since thrombin rapidly activated Src in VSMC, Src may be involved in the cross-talk between the G protein-coupled receptor agonist and a tyrosine kinase receptor such as IGF-1R. PMID- 7499265 TI - ST2/T1 protein functionally binds to two secreted proteins from Balb/c 3T3 and human umbilical vein endothelial cells but does not bind interleukin 1. AB - The ST2/T1 receptor, a homologue of the interleukin 1 receptor (IL-1R), was expressed in COS and Drosophila S2 cells as a human IgG-Fc fusion protein. While a type I IL-1RFc fusion protein bound human IL-1 in vitro, the ST2Fc fusion protein did not. Furthermore, IL-1 stimulated a synthetic interleukin-8 promoter reporter gene that was cotransfected into Jurkat cells with a full-length IL-1R type I (IL-1RI) or a chimeric receptor composed of the IL-1RI extracellular domain and ST2 intracellular domain. In contrast, IL-1 did not stimulate the interleukin-8 promoter when cotransfected with a full-length ST2 or an ST2 extracellular/IL-1R intracellular domain fusion protein. Both IL-1RI and the IL 1R/ST2R chimeric receptor also activated a receptor-associated kinase and CSBP/p38 MAP kinase. Using ST2Fc receptor, we have identified, through receptor precipitation, receptor-dot blot and surface plasmon resonance, a putative ligand of ST2 secreted from Balb/c 3T3 and human umbilical vein endothelial cells. The putative ligand was also able to stimulate CSBP/p38 MAP kinase through the ST2 receptor. These results suggest that the ST2 is not an IL-1 receptor but rather has its own cognate ligand. PMID- 7499266 TI - Identification of an I kappa B alpha-associated protein kinase in a human monocytic cell line and determination of its phosphorylation sites on I kappa B alpha. AB - Nuclear factor kappa B (NF-kappa B) is stored in the cytoplasm as an inactive form through interaction with I kappa B. Stimulation of cells leads to a rapid phosphorylation of I kappa B alpha, which is presumed to be important for the subsequent degradation. We have recently reported the establishment of a lipopolysaccharide (LPS)-dependent cell-free activation system of NF-kappa B in association with the induction of I kappa B alpha phosphorylation. In this study, we have identified a kinase in cell extracts from the LPS-stimulated human monocytic cell line, THP-1, that specifically binds and phosphorylates I kappa B alpha. LPS stimulation transiently enhanced the I kappa B alpha-bound kinase activity in THP-1 cells. Mutational analyses of I kappa B alpha and competition experiments with the synthetic peptides identified major phosphorylation sites by the bound kinase as Ser and Thr residues in the C-terminal acidic domain of I kappa B alpha. Moreover, we show that the peptide, corresponding to the C terminal acidic domain of I kappa B alpha, blocked the LPS-induced NF-kappa B activation as well as inducible phosphorylation of endogenous I kappa B alpha in a cell-free system using THP-1 cells. These results suggested that the bound kinase is involved in the signaling pathway of LPS by inducing the phosphorylation of the C-terminal region of I kappa B alpha and subsequent dissociation of the NF-kappa B.I kappa B alpha complex. PMID- 7499268 TI - Isolation of DAP3, a novel mediator of interferon-gamma-induced cell death. AB - Interaction of certain cytokines with their corresponding cell-surface receptors induces programmed cell death. Interferon-gamma induces in HeLa cells a type of cell death with features characteristic of programmed cell death. Here, we report the isolation of a novel gene, DAP3 (death-associated protein-3), involved in mediating interferon-gamma-induced cell death. The rescue of this gene was performed by a functional selection approach of gene cloning that is based on transfection with an antisense cDNA expression library. The antisense RNA mediated inactivation of the DAP3 gene protected the cells from interferon-gamma induced cell death. This property endowed the cells expressing it with a growth advantage in an environment restrictive due to the continuous presence of interferon-gamma and thus provided the basis of its selection. The gene is transcribed into a single 1.7-kilobase mRNA, which is ubiquitously expressed in different tissues and codes for a 46-kDa protein carrying a potential P-loop motif. Ectopic expression of DAP3 in HeLa cells was not compatible with cell growth, resulting in a 16-fold reduction in the number of drug-resistant stable clones. The data presented suggest that DAP3 is a positive mediator of cell death induced by interferon-gamma. PMID- 7499267 TI - Triggering of the human interleukin-6 gene by interferon-gamma and tumor necrosis factor-alpha in monocytic cells involves cooperation between interferon regulatory factor-1, NF kappa B, and Sp1 transcription factors. AB - We investigated the molecular basis of the synergistic induction by interferon gamma (IFN-gamma)/tumor necrosis factor-alpha (TNF-alpha) of human interleukin-6 (IL-6) gene in THP-1 monocytic cells, and compared it with the basis of this induction by lipopolysaccharide (LPS). Functional studies with IL-6 promoter demonstrated that three regions are the targets of the IFN-gamma and/or TNF-alpha action, whereas only one of these regions seemed to be implicated in LPS activation. The three regions concerned are: 1) a region between -73 and -36, which is the minimal element inducible by LPS or TNF-alpha; 2) an element located between -181 and -73, which appeared to regulate the response to IFN-gamma and TNF-alpha negatively; and 3) a distal element upstream of -224, which was inducible by IFN-gamma alone. LPS signaling was found to involve NF kappa B activation by the p50/p65 heterodimers. Synergistic induction of the IL-6 gene by IFN-gamma and TNF-alpha, in monocytic cells, involved cooperation between the IRF 1 and NF kappa B p65 homodimers with concomitant removal of the negative effect of the retinoblastoma control element present in the IL-6 promoter. This removal occurred by activation of the constitutive Sp1 factor, whose increased binding activity and phosphorylation were mediated by IFN-gamma. PMID- 7499269 TI - Transient accumulation of heme O (cytochrome o) in the cytoplasmic membrane of semi-anaerobic Anacystis nidulans. Evidence for oxygenase-catalyzed heme O/A transformation. AB - Incubation of obligately photoautotrophic and aerobic cyanobacterium Anacystis nidulans (Synechococcus sp. PCC 6301) in the light in the presence of the photo system II inhibitor 3-(3,4-dichlorophenyl)-1,1-dimethylurea and equilibrated with approximately 1% (v/v) O2 in N2 (10 microM O2 in solution) led to a decrease of the heme A content of isolated cytoplasmic membranes and to the appearance of heme O. The latter was not seen in membranes from fully aerated cells (> 210 microM dissolved O2). Non-covalently bound hemes extracted from the membranes were identified by reversed phase high performance liquid chromatography. Heme A and O contents of the membranes changed in a reversible fashion solely depending on the ambient oxygen regime. Both hemes A and O combine with the same apoprotein as suggested by immunoblotting. CO/reduced-minus-reduced optical difference spectra, photoaction spectra of CO-inhibited O2 uptake by the membranes, and pyridine hemochrome spectra pointed to either heme belonging to a functional form of the terminal oxidase. The NADH:O2 oxidoreductase reaction catalyzed by membranes from both high O2 and low O2 cells was strictly dependent on the addition of catalytic amounts of cytochrome c, fully inhibited by 1.2 microM KCN, and insensitive to 5 microM 2-n-heptyl-4-hydroxyquinoline-N-oxide. O2 uptake by the membranes was effectively catalyzed by N,N,N',N'-tetramethyl-p phenylenediamine but not 2-methylnaphthoquinol or plastoquinol-1 as artificial substrates. Therefore we conclude that the cyanobacterial respiratory oxidase, irrespective of the type of heme in its O2-reducing center, is a cytochrome c rather than a quinol oxidase. PMID- 7499270 TI - The acid stabilization of plasminogen activator inhibitor-1 depends on protonation of a single group that affects loop insertion into beta-sheet A. AB - The serpin plasminogen activator inhibitor-1 (PAI-1) spontaneously adopts an inactive or latent conformation by inserting the N-terminal part of the reactive center loop as strand 4 into the major beta-sheet (sheet A). To examine factors that may regulate reactive loop insertion in PAI-1, we determined the inactivation rate of the inhibitor in the pH range 4.5-13. Below pH 9, inactivation led primarily to latent PAI-1, and one predominant effect of pH on the corresponding rate constant could be observed. Protonation of a group exhibiting a pKa of 7.6 (25 degrees C, ionic strength = 0.15 M) reduced the rate of formation of latent PAI-1 by a factor of 35, from 0.17 h-1 at pH 9 to about 0.005 h-1 below pH 6. The ionization with a pKa 7.6 was found to have no effect on the rate by which PAI-1 inhibits trypsin and is therefore unlikely to change the flexibility of the loop or the orientation of the reactive center. The peptides Ac-TEASSSTA and Ac-TVASSSTA (cf. P14-P7 in the reactive loop of PAI-1) formed stable complexes with PAI-1 and converted the inhibitor to a substrate for tissue type plasminogen activator. We found that peptide binding and formation of latent PAI-1 are mutually exclusive events, similarly affected by the pKa 7.6 ionization. This is direct evidence that external peptides can substitute for strand 4 in beta-sheet A of PAI-1 and that the pKa 7.6 ionization regulates insertion of complementary, internal or external, strands into this position. A model that accounts for the observed pH effects is presented, and the identity of the ionizing group is discussed based on the structure of latent PAI-1. The group is tentatively identified as His-143 in helix F, located on top of sheet A. PMID- 7499271 TI - A novel promoter for vascular endothelial growth factor receptor (flt-1) that confers endothelial-specific gene expression. AB - The human transmembrane fms-like receptor tyrosine kinase Flt-1 is one of the receptors for vascular endothelial growth factor, a growth factor which induces endothelial proliferation and vascular permeability. Flt-1 is expressed specifically in endothelium and is likely to play a role in tumor angiogenesis and embryonic vascularization. To elucidate the molecular basis for the endothelial specific expression of Flt-1, the promoter region has been isolated and functionally characterized. The promoter region contains a TATA box, a GC rich region, and putative transcription factor binding elements such as cAMP response element binding protein/activating transcription factor (CREB/ATF) and ets. Adenovirus-mediated transient expression of the flt-1 promoter/luciferase fusion gene in endothelial cells and other cell types demonstrated that a 1 kilobase fragment of the 5'-flanking region of flt-1 is involved in the endothelial-specific expression. A CREB/ATF element was found to be essential for basal transcription of the flt-1 expression. In addition, we also showed that the first intron negatively regulates flt-1 promoter activity. The flt-1 promoter will be useful in functional studies on the regulation of endothelial-specific gene expression and also as a tool in targeting the expression of exogenously introduced genes to the endothelium. PMID- 7499272 TI - The human urinary epidermal growth factor (EGF) precursor. Isolation of a biologically active 160-kilodalton heparin-binding pro-EGF with a truncated carboxyl terminus. AB - In this report, we describe the isolation from human urine of a predominant 160 kDa epidermal growth factor (EGF)-immunoreactive glycoprotein that exhibits affinity for heparin. The purification procedure involved concentration and dialysis of 20-30-liter batches of fresh urine on a high capacity ultrafiltration apparatus followed by chromatography on DEAE-Sephacel, heparin-agarose, and Sephacryl S-300. A nearly homogeneous preparation of 160-kDa protein was obtained with a yield of approximately 1 mg of 160-kDa protein from 25 liters of urine. The amino-terminal sequence of the purified 160-kDa protein, H2N-SAPQHXSXPEGTXA-, matched residues 21-34 of the predicted sequence of human prepro-EGF and established that the 160 kDa protein (pro-EGF) is a product of the prepro-EGF gene. Characterization of the carboxyl terminus of the purified protein by digestion with carboxypeptidase B and by immunoblotting with antisera against synthetic carboxyl-terminal and juxtatransmembrane peptides of prepro-EGF indicated that the carboxyl terminus has been truncated at an arginine residue that corresponds, most likely, to the carboxyl-terminal arginine of the EGF moiety. The intact 160-kDa pro-EGF is biologically active as evidenced by its specific binding to the EGF receptor and activation of the EGF receptor tyrosine kinase in A-431 cell membranes. Purified pro-EGF competitively inhibited the binding of 125I-EGF to human fibroblasts, and it stimulated the proliferation of these cells in culture. When immobilized onto culture dishes, the heparin-binding pro-EGF appeared to function both as an adhesion molecule and as a growth factor for serum-free mouse embryo cells. PMID- 7499274 TI - Properties of Acanthamoeba myosin I heavy chain kinase bound to phospholipid vesicles. AB - The actin-activated Mg(2+)-ATPase and in vitro motility activities of the three Acanthamoeba myosin I isozymes depend upon phosphorylation of their single heavy chains by myosin I heavy chain kinase. Previously, the kinase had been shown to be activated by autophosphorylation, which is enhanced by acidic phospholipids, or simply by binding to purified plasma membranes in the absence of significant autophosphorylation. In this paper, we show that the rate of phosphorylation of myosin I by unphosphorylated kinase is approximately 20-fold faster when both the myosin I and the kinase are bound to acidic phospholipid vesicles than when both are soluble. This activation is not due to an increase in the local concentrations of vesicle-bound kinase and myosin I. Thus, acidic phospholipids, like membranes, can activate myosin I heavy chain kinase in the absence of significant autophosphorylation, i.e. membrane proteins are not required. Kinetic studies show that both binding of kinase to phospholipid vesicles and autophosphorylation of kinase in the absence of phospholipid increase the Vmax relative to soluble, unphosphorylated kinase with either an increase in the apparent Km (when myosin I is the substrate) or no significant change in Km (when a synthetic peptide is the substrate). Kinetic data showed that autophosphorylation of phospholipid-bound kinase is both intermolecular and intervesicular, and that phosphorylation of phospholipid-bound myosin I by phospholipid-bound kinase is also intervesicular even when the kinase and myosin are bound to the same vesicles. The relevance of these results to the activation of myosin I heavy chain kinase and phosphorylation of myosin I isozymes in situ are discussed. PMID- 7499273 TI - Imidazoline/guanidinium binding domains on monoamine oxidases. Relationship to subtypes of imidazoline-binding proteins and tissue-specific interaction of imidazoline ligands with monoamine oxidase B. AB - Pharmacologically active compounds with an imidazoline and/or guanidinium moiety are recognized with high affinity by a family of membrane-bound proteins collectively known as imidazoline binding sites or imidazoline/guanidinium receptive sites. Two such receptive sites may correspond to imidazoline binding domains identified on the A and B isoforms of monoamine oxidase (MAO), but the detection of monoamine oxidase isoforms in multiple tissues contrasts with the restricted expression of imidazoline-binding proteins. To address these issues, we determined the relationship between monoamine oxidase isoforms and subtypes of imidazoline-binding proteins in human tissues known to express one or both isoforms of MAO. 2-(3-Azido-4-[125I]iodophenoxy)methylimidazoline ([125I]A-ZIPI), a photoaffinity adduct that selectively labels imidazoline-binding proteins, photolabeled an M(r) = approximately 59,000 peptide in liver and an M(r) = approximately 63,000 peptide in placenta, consistent with the M(r) of the MAO isoforms identified by immunoblots in these tissues. The photolabeled species in liver was immunoprecipitated with MAO-B selective antibodies, whereas the photolabeled species in placenta was immunoprecipitated by MAO-A selective antibodies consistent with the isoform of MAO predominantly expressed in these tissues. The imidazoline/guanidinium ligands interact with the enzyme at a site distinct from the substrate recognition domain, and the immunoprecipitated peptides in liver and placenta display distinct ligand recognition properties consistent with those reported for subtypes of imidazoline binding sites. However, the imidazoline binding domain was not detected in platelet membrane preparations containing amounts of MAO-B equivalent to those in the photolabeled liver membranes indicating that recognition of this domain is tissue-restricted. Restricted access to the imidazoline binding domain on platelet MAO-B was not altered by membrane washing with 500 mM KCl or by solubilization and partial purification of the enzyme suggesting that there are distinct subpopulations of MAO. Identification of a binding domain on MAO that recognizes this class of pharmacologically active compounds suggests a novel mechanism for regulation of substrate oxidation/selectivity or that the enzyme may subserve an as yet undefined function. PMID- 7499275 TI - Both the amino and carboxyl termini of Dictyostelium myosin essential light chain are required for binding to myosin heavy chain. AB - Dictyostelium myosin deficient in the essential light chain (ELC) does not function normally either in vivo or in vitro (Pollenz, R. S., Chen, T. L., Trivinos-Lagos, L., and Chisholm, R. L. (1992) Cell 69, 951-962). Since normal myosin function requires association of ELC, we investigated the domains of ELC that are necessary for binding to the myosin heavy chain (MHC). Deleting the NH2 terminal 11 or 28 amino acid residues (delta N11 or delta N28) or the COOH terminal 15 amino acid residues (delta C15) abolished binding of the ELC to the MHC when the mutants were expressed in wild-type (WT) cells. In contrast, the ELC carrying deletion or insertion of four amino acid residues (D4 or I4) in the central linker segment bound the MHC in WT cells, although less efficient competition with WT ELC suggested that the affinity for the MHC is reduced. When these mutants were expressed in ELC-minus (mlcE-) cells, where the binding to the heavy chain is not dependent on efficient competition with the endogenous ELC, delta N28 and delta N11 bound to the MHC at 15% of WT levels and delta C15 did not bind to a significant degree. I4 and D4, however, bound with normal stoichiometry. These data indicate that residues at both termini of the ELC are required for association with the MHC, while the central linker domain appears to be less critical for binding. When the mutants were analyzed for their ability to complement the cytokinesis defect displayed by mlcE- cells, a correlation to the level of ELC carried by the MHC was observed, indicating that a stoichiometric ELC-MHC association is necessary for normal myosin function in vivo. PMID- 7499276 TI - Concordant loss of imprinting of the human insulin-like growth factor II gene promoters in cancer. AB - The human insulin-like growth factor II (IGFII) gene has been shown to be imprinted for the promoters P2, P3, and P4 but not for the promoter P1 in liver and chondrocytes. Loss of imprinting of the IGFII gene has been found in a variety of human tumors including rhabdomyosarcoma and lung cancer. In this report, we determined whether loss of imprinting in tumors displays a promoter specific pattern. We examined allelic expression of all four IGFII promoters in rhabdomyosarcoma, lung cancer, and normal skeletal muscle. We demonstrate that the imprinting of all IGFII promoters is relaxed in rhabdomyosarcoma and lung cancer. These data suggest that loss of imprinting of IGFII gene promoters may be regulated coordinately by a common mechanism in these tumors. Unexpectedly, we also found that P1, in addition to P2, P3, and P4 is monoallelically expressed in three informative adult skeletal muscle tissues. This indicates that imprinting of the IGFII promoter P1 occurs in a tissue-specific manner. PMID- 7499277 TI - Selective regulation of Lyn tyrosine kinase by CD45 in immature B cells. AB - It has been well established that protein-tyrosine phosphatase CD45 is critically involved in the regulation of initial tyrosine phosphorylation and effector functions of T and B cells. However, the signaling pathway governed by CD45 is not completely understood. In B cells, it has not been unequivocally resolved as to which protein-tyrosine kinases (PTKs) associated with B cell antigen receptor are regulated by CD45 in intact cells. As a first step toward the elucidation of CD45-initiated signaling events, we have tried to identify physiological substrates for CD45 by analyzing PTK activity in CD45-deficient clones recently generated from the immature B cell line WEHI-231. The results clearly demonstrated that among PTKs examined (Lyn, Lck, and Syk), only Lyn kinase is dysregulated in the absence of CD45 such that without B cell antigen receptor ligation, Lyn is hyperphosphorylated and activated in CD45-negative clones. Thus, Lyn seems to be a selective in vivo substrate for CD45 in immature B cells. PMID- 7499278 TI - Insulin-stimulated GLUT4 translocation is mediated by a divergent intracellular signaling pathway. AB - Insulin stimulates glucose transport largely by mediating translocation of the insulin-sensitive glucose transporter (GLUT4) from an intracellular compartment to the plasma membrane. Using single cell microinjection of 3T3-L1 adipocytes, coupled with immunofluorescence detection of GLUT4 proteins, we have determined that inhibition of endogenous p21ras or injection of oncogenic p21ras has no effect on insulin-stimulated GLUT4 translocation. On the other hand, microinjection of anti-phosphotyrosine antibodies or inhibition of endogenous phosphatidylinositol 3-kinase by microinjection of a GST-p85 SH2 fusion protein markedly inhibits this biologic effect of insulin. These data suggest that the p21ras/mitogen-activated protein kinase pathway is not involved in this metabolic effect of insulin, whereas tyrosine phosphorylation and stimulation of phosphatidylinositol 3-kinase activity are critical components of this signaling pathway. PMID- 7499279 TI - Cdc42 and PAK-mediated signaling leads to Jun kinase and p38 mitogen-activated protein kinase activation. AB - The PAK family of protein kinases has been suggested as a potential target of the Cdc42 and Rac GTPases based on studies in vitro. We show that PAK-3 is activated by Cdc42 in vivo. Both, activated (GTPase-defective) Cdc42 and a constitutively active PAK-3 mutant stimulated the activity of Jun kinase 1 (JNK1) in transfected cells. Activated Cdc42 also stimulated the activity of the related p38 mitogen activated protein kinase but was a less effective activator of ERK2. The effect of Cdc42 on JNK activity was similar to that of the potent inflammatory cytokine interleukin-1 (IL-1). The observation that a dominant-negative Cdc42 mutant inhibited IL-1 activation of JNK1 indicates a role for Cdc42 in IL-1 signaling. These results suggest that Cdc42 and PAK may mediate the effects of cytokines on transcriptional regulation. PMID- 7499280 TI - Interleukin-11 induces complex formation of Grb2, Fyn, and JAK2 in 3T3L1 cells. AB - Previous studies suggested that interleukin-11 (IL-11) induces the activation of mitogen-activated protein kinase (MAPK) in mouse 3T3L1 cells. However, the mechanisms by which IL-11 activates MAPK remain elusive. Our present results show that IL-11 promotes the formation of the active GTP-bound form of Ras, suggesting that IL-11 actions may be transduced in part through the Ras/MAPK signaling pathway. By immunoblotting and immunoprecipitation, we further demonstrate the association of tyrosine phosphoproteins with Grb2, an adaptor protein serving as a key intermediate for Ras activation. These phosphotyrosine-containing proteins have been subsequently identified to be JAK2, Fyn, and Syp. JAK2 and Fyn are transiently associated with Grb2 upon stimulation with IL-11, suggesting that JAK2 and Fyn may be involved in transducing signals from the IL-11 receptor glycoprotein 130 to the Ras system through Grb2. Taken together, these results suggest that IL-11-induced interactions of JAK2, Fyn, and Grb2 may not only provide a novel mechanism for the activation of the Ras/MAPK system but also indicate cross-talk among diverse signaling pathways. PMID- 7499281 TI - Termination as a factor in "quality control" during ribosome biogenesis. AB - In eukaryotes, nascent rDNA and 5 S rRNA gene transcripts undergo 3'-end processing after termination. Mutations in which terminator sequences in these ribosomal RNA genes are deleted completely result in highly unstable transcripts, which are not properly processed and integrated into stable ribosome structure. Mutations that retard RNA processing by extending the 3' external transcribed spacer or by introducing additional secondary structure in the spacers have a similar effect on stable transcript integration. The results indicate that proper termination coupled with efficient rRNA processing acts as a "quality control" process, which helps to ensure that only normal rRNA precursors are effectively processed and assembled into active ribosomes. PMID- 7499283 TI - Signal transduction by the formyl peptide receptor. Studies using chimeric receptors and site-directed mutagenesis define a novel domain for interaction with G-proteins. AB - The binding of small peptide ligands to high affinity chemoattractant receptors on the surface of neutrophils and monocytes leads to activation of heterotrimeric G-proteins, stimulation of phosphatidylinositol-phospholipase C (PI-PLC), and subsequently to the inflammatory response. It was recently shown (Amatruda, T. T., Gerard, N. P., Gerard, C., and Simon, M. I. (1993) J. Biol. Chem. 268, 10139 10144) that the receptor for the chemoattractant peptide C5a specifically interacts with G alpha 16, a G-protein alpha subunit of the Gq class, to trigger ligand-dependent stimulation of PI-PLC in transfected cells. In order to further characterize this chemoattractant peptide signal transduction pathway, we transfected cDNAs encoding the formylmethionylleucylphenylalanine receptor (fMLPR) into COS cells and measured the production of inositol phosphates. Ligand dependent activation of PI-PLC was seen in COS cells transfected with the fMLPR and G alpha 16 and stimulated with fMLP but not in cells transfected with receptor alone or with receptor plus G alpha q. Chimeric receptors in which the N terminal extracellular domain, the second intracellular domain, or the intracellular C-terminal tail of the fMLP receptor was replaced with C5a receptor domains (Perez, H. D., Holmes, R., Vilander, L. R., Adams, R. R., Manzana, W., Jolley, D., and Andrews, W. H. (1993) J. Biol. Chem. 268, 2292-2295) were capable of ligand-dependent activation of PI-PLC when co-transfected with G alpha 16. A chimeric receptor exchanging the first intracellular domain of the fMLPR was constitutively activated, stimulating PI-PLC in the absence of ligand. Constitutive activation of PI-PLC, to a level 233% of that seen in cells transfected with wild-type fMLP receptors, was dependent on G alpha 16. Site directed mutagenesis of the first intracellular domain of the fMLPR (amino acids 54-62) reveals this to be a domain necessary for ligand-dependent activation of G alpha 16. These results suggest that different receptors which mediate similar biochemical responses may utilize distinct mechanisms to activate G-proteins. Differences among the signaling pathways triggered by chemoattractant factor receptors suggest an opportunity for pharmacologic modifications of the inflammatory response. PMID- 7499282 TI - The essential function of protein-disulfide isomerase is to unscramble non-native disulfide bonds. AB - Protein-disulfide isomerase (PDI) is an abundant protein of the endoplasmic reticulum that catalyzes dithiol oxidation and disulfide bond reduction and isomerization using the active site CGHC. Haploid pdi1 delta Saccharomyces cerevisiae are inviable, but can be complemented with either a wild-type rat PDI gene or a mutant gene coding for CGHS PDI (shufflease). In contrast, pdi1 delta yeast cannot be complemented with a gene coding for SGHC PDI. In vitro, shufflease is an efficient catalyst for the isomerization of existing disulfide bonds but not for dithiol oxidation or disulfide bond reduction. SGHC PDI catalyzes none of these processes. These results indicate that in vivo protein folding pathways contain intermediates with non-native disulfide bonds, and that the essential role of PDI is to unscramble these intermediates. PMID- 7499284 TI - N51 competes 125I-interleukin (IL)-8 binding to IL-8R beta but not IL-8R alpha. Structure-function analysis using N51/IL-8 chimeric molecules. AB - We have demonstrated that the mouse chemokine N51, also known as KC, can compete for 125I-human interleukin-8 (IL-8) binding to NIH 3T3 cells expressing the human IL-8 receptor beta (NIH-IL-8R beta) but not the IL-8 receptor alpha (NIH-IL-8R alpha). In addition, we used the chimeras between N51 and IL-8 described previously (Heinrich, J. N., O'Rourke, E. C., Chen, L., Gray, H., Dorfman, K. S., and Bravo, R. (1994) Mol. Cell. Biol. 14, 2849-2861; Heinrich, J. N., and Bravo, R. (1995) J. Biol. Chem. 270, 4987-4989) to evaluate possible contributions of equivalent domains from each chemokine to binding and specificity. Specifically, the amino acid sequences between cysteines 2 and 3 or between cysteines 3 and 4 or the alpha-helical C-terminal end (domains I, II, and III, respectively) of one of the chemokines was exchanged with the corresponding sequence of the other and vice versa. Chimeras of IL-8 containing either domain II or III of N51 behaved similarly, but not identically, to IL-8 in competing 125I-IL-8 binding with both NIH-IL-8R alpha cells and NIH-IL-8R beta cells. The IL-8 chimera containing domain I of N51 did not compete. On the other hand, N51 competes 125I-IL-8 binding with NIH-IL-8R beta but not NIH-IL-8R alpha cells. The N51 chimera containing domain I of IL-8 was an agonist with NIH-IL-8R alpha cells and was an even more potent agonist with NIH-IL-8R beta cells. On the latter cells it was more potent than either IL-8 or N51. The N51 chimera containing domain II of IL 8, compared with N51, was a partial agonist with NIH-IL-8R alpha cells but was equivalent to N51 with NIH-IL-8R beta cells. However, N51 chimera containing domain III of IL-8 was a partial agonist with both cells. The results are consistent with the observations we originally made with human neutrophils and the NIH mouse IL-8R beta cells, i.e. domain I confers binding specificity for IL 8 and domains II and III of IL-8 and N51 may be interchangeable but they are not equivalent. Although we originally hypothesize that domains II and III confer binding specificity to N51, these results emphasize the role of domain III. PMID- 7499285 TI - Increased drug affinity as the mechanistic basis for drug hypersensitivity of a mutant type II topoisomerase. AB - Altered sensitivity of topoisomerase II to anticancer drugs profoundly affects the response of eukaryotic cells to these agents. Therefore, several approaches were employed to elucidate the mechanism of drug hypersensitivity of the mutant yeast type II topoisomerase, top2H1012Y. This mutant, which is approximately 5 fold hypersensitive to ellipticine, formed DNA cleavage complexes more rapidly than the wild-type yeast enzyme in the presence of the drug. Conversely, no change in the rate of DNA religation was observed. There was, however, a correlation between increased cleavage rates and enhanced drug binding affinity. The apparent dissociation constant for ellipticine in the mutant topoisomerase II.drug.DNA ternary complex was approximately 5-fold lower than in the wild-type ternary complex. Furthermore, the apparent KD value for the mutant binary (topoisomerase II.drug) complex was approximately 2-fold lower than the corresponding wild-type complex, indicating that drug hypersensitivity is intrinsic to the enzyme. These findings strongly suggest that the enhanced ellipticine binding affinity for topoisomerase II is the mechanistic basis for drug hypersensitivity of top2H1012Y. PMID- 7499286 TI - Specificity in recognition of amyloid-beta peptide by the serpin-enzyme complex receptor in hepatoma cells and neuronal cells. AB - The serpin-enzyme complex (SEC) receptor was originally identified using a synthetic peptide (peptide 105Y) based on the sequence of a candidate receptor binding domain of alpha 1-antitrypsin (1-AT) and was subsequently shown to be a receptor on the surface of hepatocytes, monocytes, and neutrophils for recognition of alpha 1-AT-elastase and several other serpin-enzyme complexes (Perlmutter, D. H., Glover, G. I., Rivetna, M., Schasteen, C. S., and Fallon, R.J. (1990) Proc. Natl. Acad. Sci. U.S.A. 87, 3753-3757). Studies of the minimal requirements for binding to SEC receptor (SEC-R) showed that a pentapeptide FVFLM within the carboxyl-terminal tail of alpha 1-AT was sufficient for binding to SEC R and interacted with SEC-R in a sequence-specific manner (Joslin, G., Krause, J. E., Hershey, A. D., Adams, S. P., Fallon, R. J., and Perlmutter, D. H. (1991) J. Biol. Chem. 266, 21897-21902). Sequence motifs bearing homology with this pentapeptide domain were found in the amyloid-beta peptide, and amyloid-beta peptide 1-42 was shown to compete for binding to SEC-R on hepatoma cells (Joslin, G., Fallon, R. J., Bullock, J., Adams, S. P., and Perlmutter, D. H. (1991) J. Biol. Chem, 266, 11281-11288). In this study we examined the sequence specificity by which amyloid-beta peptide competes for binding to SEC-R and examined the possibility that SEC-R is expressed in cells of neuronal origin. The results show that amyloid-beta-(25-35) and amyloid-beta-(31-35) compete for binding to SEC-R as effectively as amyloid-beta-(1-39), amyloid-beta-(1-40), and amyloid-beta-(1 42). Amyloid-beta-(1-16) does not compete for binding to SEC-R. There is cross competition for binding to the same site by 125I-peptide 105Y and amyloid-beta (25-35) as well as by 125I-Y amyloid-beta-(25-35) and peptide 105Y. By deletions and substitutions within amyloid-beta-(25-35) and generation of chimeric amyloid beta-alpha 1-AT peptides, amyloid-beta-(31-35) is shown to be critical for binding to the SEC receptor. However, the upstream region, amyloid-beta-(25-30), also contributes to recognition by SEC-R. The SEC-R is present on the surface of a neuronal cell line PC12 as well as that of murine cortical neurons in primary culture, and the specificity of neuronal SEC-R for amyloid-beta peptide is identical to that on hepatoma cells. Finally SEC-R mediates internalization and degradation of amyloid beta-peptide in PC12 cells.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 7499287 TI - Cyclic AMP-insensitive activation of c-Src and Syk protein-tyrosine kinases through platelet membrane glycoprotein VI. AB - Platelet glycoprotein (GP) VI is a so-far uncharacterized 62-kDa membrane protein, whose deficiency results in selective impairment in collagen-induced platelet aggregation. Our group previously reported a human polyclonal antibody (anti-p62 IgG) that induces activation of normal, but not of GPVI-deficient, platelets in an Fc-independent manner. The F(ab')2 fragments of this antibody (F(ab')2-anti-p62) stimulated tyrosine phosphorylation of numerous proteins, which was not prevented even in the presence of cAMP-increasing agents such as prostacyclin. Pretreatment of platelets with the protein-tyrosine kinase (PTK) inhibitor tyrphostin A47 completely abolished F(ab')2-anti-p62-induced platelet aggregation in parallel with dose-dependent inhibition of protein-tyrosine phosphorylation, indicating an essential requirement of PTK activity for generating GPVI-mediated signaling. We found that two cytosolic PTKs, c-Src and Syk, became rapidly activated in response to F(ab')2-anti-p62 in a way insensitive to elevation of cAMP. In contrast, in the presence of prostacyclin, F(ab')2-anti-p62 did not stimulate tyrosine phosphorylation of the focal adhesion kinase. cAMP-insensitive activation of c-Src and Syk was also observed in collagen but not thrombin-stimulated platelets. Moreover, either F(ab')2-anti-p62 or collagen stimulated cAMP-insensitive tyrosine phosphorylation of phospholipase C-gamma 2. These results indicate that the receptor-mediated activation of several PTKs in platelets is regulated through a cAMP-sensitive or -insensitive mechanism depending on the nature of each stimulus, and also suggest that GPVI engagement is coupled to cAMP-insensitive activation of c-Src and Syk accompanied by tyrosine phosphorylation of numerous substrates including phospholipase C gamma 2 in a manner similar to collagen stimulation. PMID- 7499288 TI - An anti-CD3 single-chain immunotoxin with a truncated diphtheria toxin avoids inhibition by pre-existing antibodies in human blood. AB - Diphtheria toxin (DT) is often used in the construction of immunotoxins. One potential problem using DT-based immunotoxins is the pre-existing anti-DT antibodies present in human blood due to vaccination. The present study examined the effect of human serum with pre-existing anti-DT antibodies on the toxicity of UCHT1-CRM9, an immunotoxin directed against CD3 molecules on T-lymphocytes. Sera with detectable anti-DT antibodies at 1:100 or greater dilutions inhibited the immunotoxin toxicity. Experiments with radio-labeled UCHT1-CRM9 indicate that anti-DT antibodies partially block its binding to the cell surface as well as inhibit the translocation from the endosome to the cytosol. The inhibitory effect could be adsorbed using a full-length DT mutant or B-subfragment. A C-terminal truncation mutant could not adsorb the inhibitory effect, suggesting that the last 150 amino acids contain the epitope(s) recognized by the inhibitory antibodies. Therefore, an anti-CD3 single-chain immunotoxin, sFv-DT390, was made with a truncated DT. The IC50 of sFv-DT390 was 4.8 x 10(-11) M, 1/16 the potency of the divalent UCHT1-CRM9. More importantly, sFv-DT390 toxicity was only slightly affected by the anti-DT antibodies in human sera. PMID- 7499289 TI - Uncoupling oxygen transfer and electron transfer in the oxygenation of camphor analogues by cytochrome P450-CAM. Direct observation of an intermolecular isotope effect for substrate C-H activation. AB - The hydroxylation of (1R)-camphor by cytochrome P450-CAM involves almost complete coupling of electron to oxygen transfer. Modifications at C-5 of camphor, the normal site of hydroxylation by P450-CAM, lead to as much as 98% uncoupling of electron and oxygen transfer as well as to decreases in the rate of electron uptake (up to 10-fold) and the rate of oxygenated product formation (up to 210 fold). Two modes of uncoupling are seen: (a) two-electron uncoupling in which the decrease in oxygenated product formation is balanced by increases in H2O2 formation and (b) four-electron "oxidase" uncoupling where the NADH/O2 ratio has changed from one to nearly two and relatively little H2O2 is formed. Both enantiomers of 5-methylenylcamphor are two-electron uncouplers, while (1R)- and (1S)-5,5-difluorocamphor and (1R)-9,9,9-d3-5,5-difluorocamphor are four-electron uncouplers. An intermolecular isotope effect of 11.7 is observed for oxygenation of C-9 in (1R)-5,5-difluorocamphor. With this substrate, the significant decrease in the rate of oxygenated product formation combined with the large isotope effect suggest that the rate-limiting step has switched from electron to oxygen transfer. PMID- 7499292 TI - Mass and positional isotopomer analysis of glucose metabolism in periportal and pericentral hepatocytes. AB - To determine whether the source of carbon for the indirect pathway of hepatic glycogen synthesis differs between the periportal and pericentral zones, we studied seven 24-h-fasted conscious rats given a constant 2-h intraduodenal infusion of glucose, 40% labeled with [U-13C]glucose (99% 13C enriched), to raise and maintain plasma glucose concentration at approximately 10 mM. Glycogen, glutamate, aspartate, and alanine were selectively sampled from the periportal and pericentral zones of the liver by the dual-digitonin pulse technique and analyzed by 13C-NMR for positional isotopomer distribution and by gas chromatography-mass spectrometry for mass isotopomer distribution. Plasma glucose mass isotopomer distribution was determined from gas chromatography-mass spectrometry. The isotopomer distribution indicates that there was no significant difference between the zones with respect to 1) percent direct flux of glucose into the glycogen (periportal, 34 +/- 4; pericentral, 38 +/- 4), 2) extent of oxaloacetate/fumarate equilibration (periportal, 0.54 +/- .01;, pericentral, 0.53 +/- 0.01), 3) dilution of tracer in oxaloacetate (periportal, 0.64 +/- 0.07;, pericentral, 0.64 +/- 0.07), or 4) inflow of pyruvate versus tricarboxylic acid cycle flux (periportal, 0.70 +/- 0.20; pericentral, 0.68 +/- 0.16). Positional isotopomer populations, determined from the 13C-13C splitting in C3 and C4 of periportal and pericentral glycogen, were indistinguishable, indicating no significant differences in the source of the 3-carbon precursors for hepatic glycogen synthesis by the indirect pathway. In conclusion, glucose metabolism is the same in the periportal and pericentral zones with regard to 1) the relative flux of carbon via the direct/indirect pathways, 2) the source of the 3-carbon precursor used in the indirect pathway of glycogen synthesis, and 3) the flux of the 3-carbon precursors through the tricarboxylic acid cycle. PMID- 7499290 TI - Using modified nucleotides to map the DNA determinants of the Tus-TerB complex, the protein-DNA interaction associated with termination of replication in Escherichia coli. AB - A series of modified nucleotides was used to map the hydrogen-bonding and hydrophobic sites of the TerB DNA required for Tus interaction. Each of four consensus guanine residues in the TerB-binding site was replaced by 7 deazaguanine, 2-aminopurine, or inosine nucleobase analogues, and each thymine by a uracil analogue. The observable equilibrium dissociation constant for the Tus protein-TerB DNA complex was measured at pH 7.5, 25 degrees C, and 150 mM potassium glutamate using a competition binding method. Substitutions made at position 10 with a 7-deazaguanine, 2-aminopurine, or inosine analogue had a large effect on the stability of the complex, approximately +3 kcal/mol in each case. Substitutions made at positions 13 and 17 had a varied response. For uracil substitutions, potential hydrophobic sites were identified at six positions in the TerB DNA. The energetic penalty for the removal of a single methyl group ranged between +1 and +2 kcal/mol. Rate dissociation measurements agree with these results. Overall, major and minor groove determinants are required for binding. An unusual result was that the conserved nucleotide at position 6 did not significantly affect in vitro binding of the complex. PMID- 7499291 TI - Calcineurin feedback inhibition of agonist-evoked cAMP formation. AB - The effects of immunosuppressant blockers of calcineurin (protein phosphatase 2B) on cAMP formation and hormone release were investigated in mouse pituitary tumor (AtT20) cells. Immunosuppressants enhanced corticotropin-releasing factor- and isoproterenol-evoked cAMP production in proportion with their potency to block calcineurin. Further analysis of cAMP production revealed that intracellular Ca2+ derived through voltage-regulated calcium channels reduces cAMP formation induced by corticotropin releasing-factor or beta 2-adrenergic stimulation and that this effect of Ca2+ is inhibited by blockers of calcineurin. AtT20 cells were found to express at least three species of adenylyl cyclase mRNA-encoding types 1 and 6 as well as a novel isotype, which appeared to be the predominant species. In two cell lines expressing very low or undetectable levels of the novel cyclase mRNA (NCB20 and HEK293 cells respectively), corticotropin-releasing factor-induced cAMP formation was not altered upon blockage of calcineurin activity. These data identify calcineurin as a Ca2+ sensor that mediates the negative feedback effect of intracellular Ca2+ on receptor-stimulated cAMP production. Furthermore, the effect of calcineurin on cAMP synthesis appears to be associated with the expression of a novel adenylyl cyclase isotype, which is highly abundant in AtT20 cells. PMID- 7499293 TI - Molecular cloning of NIPP-1, a nuclear inhibitor of protein phosphatase-1, reveals homology with polypeptides involved in RNA processing. AB - NIPP-1 was originally isolated as a potent and specific nuclear inhibitory polypeptide (16-18 kDa) of protein phosphatase-1. We report here the cDNA cloning of NIPP-1 from bovine thymus and show that the native polypeptide consists of 351 residues and has a calculated mass of 38.5 kDa. The bacterially expressed central third of NIPP-1 completely inhibited the type-1 catalytic subunit, but displayed a reduced inhibitory potency after phosphorylation by protein kinase A and casein kinase 2. Translation of NIPP-1 mRNA in reticulocyte lysates resulted in the accumulation of both intact NIPP-1 and a smaller polypeptide generated by alternative initiation at the codon corresponding to Met143. A data base search showed that the COOH terminus of NIPP-1 is nearly identical to the human ard-1 protein (13 kDa), which has been implicated in RNA processing (Wang, M., and Cohen, S. N. (1994) Proc. Natl. Acad. Sci. U. S. A. 91, 10591-10595). Comparison of the cDNAs encoding ard-1 and NIPP-1 suggests that their mRNAs are generated by alternative splicing of the same pre-mRNA. Western blotting with antibodies against the COOH terminus of NIPP-1, however, showed a single polypeptide of 47 kDa, which was enriched in the nucleus. Northern analysis revealed a single transcript of 2.2 kilobases in bovine thymus and of 2.4 kilobases in various human tissues. PMID- 7499294 TI - Actin polymerization induced by GTP gamma S in permeabilized neutrophils is induced and maintained by free barbed ends. AB - To address the mechanisms through which agonists stimulate actin polymerization, we examined the roles of monomer sequestering proteins and free barbed ends on actin polymerization induced by guanosine 5'-3-O-(thio)triphosphate (GTP gamma S) in neutrophils permeabilized with streptolysin O. Addition of profilin (without GTP gamma S) caused a net decrease in F-actin. Thus, merely making profilin available in the cell was not sufficient to induce actin polymerization. On the other hand, addition of profilin hardly affected the polymerization induced by GTP gamma S, while thymosin beta 4 or DNase I decreased this polymerization. These data suggested that GTP gamma S induced polymerization by increasing the availability of barbed ends. In the presence of cytochalasin B, profilin did inhibit polymerization induced by GTP gamma S, demonstrating that GTP gamma S did not inhibit profilin's monomer sequestering ability. The F-actin induced by GTP gamma S was not limited by a time-dependent loss of G-actin or G-proteins from permeabilized cells since, following stimulation with suboptimal concentrations of GTP gamma S, addition of more GTP gamma S induced further polymerization. Barbed ends remained free after F-actin reached plateau since (a) cytochalasin B caused depolymerization of induced F-actin and (b) profilin did not depolymerize induced F-actin unless the cells were first treated with cytochalasin to cap barbed ends. The data indicate that GTP gamma S maintains an increased level of F actin by keeping at least a few barbed ends available for polymerization. PMID- 7499295 TI - Intracellular loop between transmembrane segments IV and V of cystic fibrosis transmembrane conductance regulator is involved in regulation of chloride channel conductance state. AB - The cystic fibrosis transmembrane conductance regulator (CFTR) contains two membrane-spanning domains; each consists of six transmembrane segments joined by three extracellular and two intracellular loops of different length. To examine the role of intracellular loops in CFTR channel function, we studied a deletion mutant of CFTR (delta 19 CFTR) in which 19 amino acids were removed from the intracellular loop joining transmembrane segments IV and V. This mutant protein was expressed in a human embryonic kidney cell line (293 HEK). Fully mature glycosylated CFTR (approximately 170 kDa) was immunoprecipitated from cells transfected with wild-type CFTR cDNA, while cells transfected with the mutant gene expressed only a core-glycosylated form (approximately 140 kDa). The chloride efflux rate (measured by 6-methoxyl-N-(3-sulfopropyl) quinolinium SPQ fluorescence) from cells expressing wild-type CFTR increased 600% in response to forskolin. In contrast, delta 19 CFTR-expressing cells had no significant response to forskolin. Western blotting performed on subcellular membrane fractions showed that delta 19 CFTR was located in the same fractions as delta F508 CFTR, a processing mutant of CFTR. These results suggest that delta 19 CFTR is located in the intracellular membranes, without reaching the cell surface. Upon reconstitution into lipid bilayer membranes, delta 19 CFTR formed a functional Cl- channel with gating properties nearly identical to those of the wild-type CFTR channel. However, delta 19 CFTR channels exhibited frequent transitions to a 6-picosiemens subconductance state, whereas wild-type CFTR channels rarely exist in this subconductance state. These data suggest that the intracellular loop is involved in stabilizing the full conductance state of the CFTR Cl- channel. PMID- 7499296 TI - The mitogen-activated protein kinase kinase MEK1 stimulates a pattern of gene expression typical of the hypertrophic phenotype in rat ventricular cardiomyocytes. AB - Adult mammalian ventricular cardiomyocytes are terminally differentiated cells that enlarge adaptively by hypertrophy. In this situation, genes normally expressed in the fetal ventricular cardiomyocyte (e.g. atrial natriuretic factor (ANF), beta-myosin heavy chain (beta-MHC), and skeletal muscle (SkM) alpha-actin) are re-expressed, and there is transient expression of immediate early genes (e.g. c-fos). Using appropriate reporter plasmids, we studied the effects of transfection of the constitutively active or dominant negative mitogen-activated protein kinase kinase MEK1 on ANF, beta-MHC, and SkM alpha-actin promoter activities in cultured ventricular cardiomyocytes. ANF expression was stimulated (maximally 75-fold) by the hypertrophic agonist phenylephrine in a dose-dependent manner (EC50, 10 microM), and this stimulation was inhibited by dominant negative MEK1. Cotransfection of dominant negative MEK1 with a dominant negative mitogen activated protein kinase (extracellular signal-regulated protein kinase (ERK2)) increased this inhibition. Transfection with constitutively active MEK1 constructs doubled ANF promoter activity. The additional cotransfection of wild type ERK2 stimulated ANF promoter activity by about 5-fold. Expression of beta MHC and SkM alpha-actin was also stimulated. Promoter activity regulated by activator protein-1 or c-fos serum response element consensus sequences was also increased. We conclude that the MEK1/ERK2 cascade may play a role in regulating gene expression during hypertrophy. PMID- 7499297 TI - Alkali metal ion dependence of inositol 1,4,5-trisphosphate-induced calcium release from rat cerebellar microsomes. AB - The effects of the alkali metal ions Na+, K+, Rb+, and Cs+ on ATP-dependent Ca2+ uptake, [3H]Inositol 1,4,5-trisphosphate (InsP3) binding, and quantal InsP3 induced Ca2+ release were investigated using rat cerebellar microsomes. Both the ion species and concentration affected the ability of the microsomes to support Ca2+ uptake with K+ being mot effective (3.8 nmol of Ca2+/min/mg at 100 mM K+). The order of efficacy of the other ions was as follows: K+ > Na+ > Rb+ = Cs+ >> Li+. The binding of [3H]InsP3 to cerebellar microsomes was, however, affected little by the presence of these ions. All these alkali metal ions (except Li+) supported InsP3-induced Ca2+ release at concentrations above 25 mM; however, the extent of Ca2+ release (expressed as a percent Ca2+ release compared with that released by the ionophore A23187) was dependent upon the ion species present. Again K+ was more potent than the other ions at facilitating InsP3-induced Ca2+ release (order of efficacy: K+ > Rb+ > Na+ > Cs+), although the concentration of InsP3 required to induce half-maximal Ca2+ release (IC50) was not significantly altered. Over the ion concentration range tested (25-100 mM), the extent of InsP3 induced Ca2+ release with both K+ and Rb+ increased in a linear fashion, while Na+ showed only a slight increase and Cs+ showed no increase over this range. The effect of K+ concentration on quantal Ca2+ release was to alter the extent of release rather than the IC50 InsP3 concentration. Using stopped-flow techniques, the effects of InsP3 and K+ concentrations on the kinetics of InsP3-induced Ca2+ release were shown to exhibit a monoexponential process in this microsomal preparation. The rate constants for Ca2+ release increased with InsP3 concentration (0.11 s-1 at 0.02 microM InsP3 to 0.5 s-1 at 40 microM InsP3); however, the relationship between the fractional extent of release and rate constants for release did not change in a similar way with InsP3 concentration. Although the fractional extent of Ca2+ release increased with K+ concentration, the rate constants for release over this K+ concentration range were unaffected. This observation leads us to question the role of K+ as a counter ion required for Ca2+ release, and we therefore postulate a role for K+ (and the other alkali metal ions) as a "co-factor" required for channel opening. PMID- 7499298 TI - CD45 protein-tyrosine phosphatase associates with the WW domain-containing protein, CD45AP, through the transmembrane region. AB - CD45 is a transmembrane protein-tyrosine phosphatase required for antigen receptor signaling in lymphocytes. CD45 activates the Src family protein-tyrosine kinases, p56lck and p59fyn, by dephosphorylating a negative regulatory tyrosine in the carboxyl terminus. Immunoprecipitation of CD45 precipitates p56lck and CD45AP. Although the function of CD45AP is unknown, it has been proposed to be an adapter between p56lck and CD45. To assess the ability of CD45AP to function as an adapter, we determined the regions required for the interaction with CD45 by expressing chimeric proteins in HeLa cells. CD45AP has a region similar to a potential protein-protein interaction domain, the WW domain. Surprisingly, this domain was not necessary for the association with CD45. Rather, a 40-amino acid sequence encompassing the putative transmembrane domain of CD45AP was sufficient to mediate binding to CD45. Similarly, a 39-amino acid sequence encompassing the CD45 transmembrane region was sufficient to direct the interaction with CD45AP. Expression of p56lck with CD45AP resulted in an interaction that could only be detected by in vitro kinase reaction, suggesting that the association of p56lck and CD45AP is weak. These data support a model in which CD45AP links CD45 with other proteins but not necessarily p56lck. PMID- 7499299 TI - Modified ligands to FA and FB in photosystem I. I. Structural constraints for the formation of iron-sulfur clusters in free and rebound PsaC. AB - Cysteines 14, 21, 34, 51, or 58 in PsaC of photosystem I (PS I) were replaced with aspartic acid (C21D and C58D), serine (C14S, C34S, and C51S), and alanine (C14A, C34A, and C51A). When free in solution, the C34S and C34A holoproteins contained two S = 1/2 ground state [4Fe-4S] clusters; all other mutant proteins contained [3Fe-4S] clusters and [4Fe-4S] clusters; in addition, there was evidence in C14S, C51S, C14A, and C51A for high spin (S = 3/2) [4Fe-4S] clusters, presumably in the modified site. These findings are consistent with the assignment of C14, C21, C51, and C58, but not C34, as ligands to FA and FB. The [4Fe-4S] clusters in the unmodified sites in C14S, C51S, C14A, and C51A remained highly electronegative, with Em values ranging from -495 to -575 mV. The [3Fe-4S] clusters in the modified sites were driven 400 to 450 mV more oxidizing than the native [4Fe-4S] clusters, with Em values ranging from -98 mV to -171 mV. A C14D/C51D double mutant contains [3Fe-4S] and S = 1/2 [4Fe-4S] clusters, showing that the 3Cys.1Asp motif is also able to accommodate a low spin cubane. When C34S, C34A, C14S, C51S, C14A, and C51A were rebound to P700-FX cores, electron transfer to FA/FB was regained, but functional reconstitution has not yet been achieved for C21D, C58D, or C14D/C51D. These data imply that PsaC requires two iron-sulfur clusters to refold, one of which must be a cubane. Since two [4Fe-4S] clusters are found in all reconstituted PS I complexes, the presence of two cubanes in free PsaC may be a necessary precondition for binding to P700-FX cores. PMID- 7499301 TI - Prevention of in vitro protein thermal aggregation by the Sulfolobus solfataricus chaperonin. Evidence for nonequivalent binding surfaces on the chaperonin molecule. AB - We have studied the effects of the Sulfolobus solfataricus chaperonin on the aggregation and inactivation upon heating of four model enzymes: chicken egg white lysozyme (one 14.4-kDa chain), yeast alpha-glucosidase (one 68.5-kDa chain), chicken liver malic enzyme (four 65-kDa subunits), and yeast alcohol dehydrogenase (four 37.5-kDa subunits). When the proteins were heated in the presence of an equimolar amount of chaperonin, 1) the aggregation was prevented in all solutions; 2) the inactivation profiles of the single-chain enzymes were comparable with those detected in the absence of the chaperonin, and enzyme activities were regained in the solutions heated in the presence of the chaperonin upon ATP hydrolysis (78 and 55% activity regains for lysozyme and alpha-glucosidase, respectively); 3) the inactivation of the tetrameric enzymes was completely prevented, whereas the activities decreased in the absence of the chaperonin. We demonstrate by gel filtration chromatography that the chaperonin interacted with the structures occurring during thermal denaturation of the model proteins and that the interaction with the single-chain proteins (but not that with the tetrameric proteins) was reversed upon ATP hydrolysis. The chaperonin had nonequivalent surfaces for the binding of the model proteins upon heating: the thermal denaturation intermediates of the single-chain proteins share Surfaces I, while the thermal denaturation intermediates of the tetrameric proteins share Surfaces II. ATP binding to the chaperonin induced a conformation that lacked Surfaces I and carried Surfaces II. These data support the concept that chaperonins protect native proteins against thermal aggregation by two mechanistically distinct strategies (an ATP-dependent strategy and an ATP independent strategy), and provide the first evidence that a chaperonin molecule bears functionally specialized surfaces for the binding of the protein substrates. PMID- 7499302 TI - Nerve growth factor binding site on TrkA mapped to a single 24-amino acid leucine rich motif. AB - The extracellular domains of the TrkA nerve growth factor (NGF) receptor and its homologs harbor a modular mosaic of potential ligand binding motifs, namely two immunoglobulin (Ig)-like modules and an LRM3 cassette consisting of a tandem array of three leucine-rich motifs (LRMs) flanked by cysteine-rich clusters (Schneider, R., and Schweiger, M. (1991) Oncogene 6, 1807-1811). Identification of a structural motif capable of specifically recognizing the various neurotrophins was achieved by assessing their affinities to isolated recombinant modules of TrkA and TrkB. In both receptors the LRM3 cassette alone could mediate the respective neurotrophin selectivities and affinities. Further tracking down of this NGF-binding site in TrkA strikingly revealed that a single LRM of 24 amino acids could bind NGF selectively with nanomolar affinity. Since this is the first example of a single LRM with a highly specific, well defined function, it might serve as a valuable tool to elucidate the general structural requirements of substrate recognition and high affinity binding in the large superfamily of LRM-containing proteins. PMID- 7499300 TI - Modified ligands to FA and FB in photosystem I. II. Characterization of a mixed ligand [4Fe-4S] cluster in the C51D mutant of PsaC upon rebinding to P700-Fx cores. AB - A Photosystem I (PS I) complex reconstituted with PsaC-C51D (aspartate in lieu of cysteine in position 51) shows light-induced EPR signals with g values, line widths, and photoreduction behavior characteristic of FB. Contrary to an earlier report, a [3Fe-4S] cluster was not located in the reconstituted PS I complex. Instead, a second set of resonances with g values of 2.044, 1.942, and 1.853 becomes EPR-visible when the C51D-PS I complex is measured at 4.2 K. This fast relaxing center, termed FA' is likely to represent a [4Fe-4S] cluster in the mixed ligand (3Cys.1Asp) site. Redox studies show that the Em of FA' and FB are 630 mV and -575 mV, respectively. Room temperature optical studies support the presence of two functioning electron acceptors subsequent to Fx, and NADP+ photoreduction rates mediated by ferredoxin and flavodoxin are nearly equivalent to the wild type. In addition to [3Fe-4S] clusters and S = 1/2 ground state [4Fe 4S] clusters, the free PsaC-C51D protein shows resonances near g = 5.5, which may represent a population of high spin (S = 3/2) [4Fe-4S] clusters in the mixed ligand FA' site. Similar to the C14D-PS I mutant complex, it is proposed that the P700-Fx core selectively rebinds those free PsaC-C51D proteins that contain two [4Fe-4S] clusters. These studies show that primary photochemistry and electron transfer rates in PS I are relatively unaffected by the presence of a highly reducing, mixed ligand cluster in the FA' site. PMID- 7499303 TI - A functional initiator element in the human beta-globin promoter. AB - Core promoters are defined by the presence of either a TATA box at approximately 30 base pairs upstream of the transcriptional start site (+1) and/or an initiator element centered around the +1 site. The prevalence, function, and significance of the various combinations of core promoter elements are as yet unclear. We describe here the identification and characterization of an initiator element in the TATA-containing human beta-globin promoter. Mutagenesis of the beta-globin initiator element at positions +2/+3 and +4/+5 abrogates transcription in a heterologous construct. Interestingly, we have found a beta-globin initiator binding activity in nuclear extracts whose presence or absence correlates with function of the beta-globin initiator. Accordingly, this binding activity may be part of the machinery required for beta-globin initiator-dependent transcription. Our analysis further describes a previously uncharacterized beta-thalassemia mutation at the +1 site as a mutation that decreases beta-globin initiator activity. Finally, consistent with other initiator elements, the beta-globin initiator requires a TFIID-containing fraction for in vitro activity. Thus, the human beta-globin promoter contains an initiator element whose function, as revealed by a beta-thalassemia mutation, is of physiological relevance. PMID- 7499305 TI - Uptake of diamidine drugs by the P2 nucleoside transporter in melarsen-sensitive and -resistant Trypanosoma brucei brucei. AB - The African trypanosome, Trypanosoma brucei brucei, possesses at least two nucleoside transporter systems designated P1 and P2, the latter being implicated in the selective uptake of melaminophenyl arsenical drugs. Since arsenical resistant trypanosomes show cross-resistance in vivo to aromatic diamidines, we have investigated whether these drugs are also substrates for the P2 nucleoside transporter. In melarsen-sensitive T. b. brucei, the diamidines, including the commonly used trypanocides, pentamidine and berenil, were found to abrogate lysis induced by the P2 transport of melarsen oxide in vitro. Measurement of [ring 3H]pentamidine transport in melarsen-sensitive T. b. brucei, demonstrated that uptake is carrier-mediated, with a Km of 0.84 microM and a Vmax of 9.35 pmol s-1 (10(8) cells)-1. Pentamidine transport appears to be P2-mediated in these cells, as pentamidine strongly inhibited uptake of [2',5',8-3H]adenosine by the P2 transporter, with a Ki of 0.56 microM. Furthermore, [ring-3H]pentamidine transport was blocked by a number of P2 transporter substrates and inhibitors, as well as by other diamidine drugs. Analysis of the uptake of pentamidine and other diamidines in melarsen-resistant trypanosomes in vitro and in vivo, which also show differential levels of resistance to these compounds in vivo, indicated that P2 transport was altered in these cells and that accumulation of these drugs was markedly reduced. PMID- 7499306 TI - Kinetics of nitrosation of thiols by nitric oxide in the presence of oxygen. AB - Nitrosothiols are powerful vasodilators. They act by releasing nitric oxide, which activates the heme protein guanylate cyclase. We have studied the kinetics of nitrosothiol formation of glutathione, cysteine, N-acetylcysteine, human serum albumin, and bovine serum albumin upon reaction with nitric oxide (NO) in the presence of oxygen. These studies have been made at low pH as well as at physiological pH. At pH 7.0, contrary to published reports, nitric oxide by itself does not react with thiols to yield nitrosothiol. However, formation of nitrosothiols is observed in the presence of oxygen. For all thiols studied, the rates of nitrosothiol formation were first order in O2 concentration and second order in NO concentration and at lower concentrations (< 5 mM thiol) also depended on thiol concentrations. Analysis of the kinetic data indicated that the rate-limiting step was the reaction of NO with oxygen. Analysis of the reaction products suggest that the main nitrosating species is N2O3: RSH+N2O3-->RSNO+NO2- + H+. Rate constants for this reaction for glutathione and several other low molecular weight thiols are in the range of 3-1.5 x 10(5) M-1 s-1, and for human and bovine serum albumins 0.3 x 10(5) M-1 s-1 and 0.06 x 10(5) M-1 s-1, respectively. The data further indicate that the reaction rate of the nitrosating species N2O3 with thiols is competitive with its rate of hydrolysis. At physiological concentrations nitrosoglutathione formation represents a significant metabolic fate of N2O3, and at glutathione concentrations of 5 mM or higher almost all of N2O3 formed is consumed in nitrosation of glutathione. Implications of these results for in vivo nitrosation of thiols are discussed. PMID- 7499304 TI - Circumvention of P-glycoprotein-mediated multiple drug resistance by phosphorylation modulators is independent of protein kinases. AB - Expression of P-glycoprotein by tumor cells confers resistance to multiple natural product drugs because of its ability to export these compounds. This transporter is a substrate for several protein kinases; however, the functional significance of its phosphorylation is not defined. We examined the effects of many activators and inhibitors of protein kinases on the activity of P glycoprotein in drug-resistant human breast carcinoma cells (MCF-7/ADR). Several phorbol esters sensitized these cells to P-glycoprotein substrate drugs; however, there was no correlation with activation of protein kinase C. The 4 alpha- and 4 beta-isomers of phorbol 12-myristate 13-acetate were equally potent in sensitizing the cells to actinomycin D and daunomycin and in increasing the intracellular accumulation of [3H]vinblastine. These effects of 4 beta-phorbol myristate acetate required much higher concentrations than were needed to increase P-glycoprotein phosphorylation and were not antagonized by staurosporine. Similar to verapamil, the phorbol esters did not sensitize MCF 7/ADR cells to cisplatin, nor parental MCF-7 cells to any of the anticancer drugs. Mezerein, K-252a, and H-89 sensitized MCF-7/ADR cells, increased intracellular accumulation of [3H]vinblastine, and antagonized photolabeling of P glycoprotein by [3H]azidopine. Therefore, phosphorylation does not appear to play a significant role in regulating P-glycoprotein activity in MCF-7/ADR cells. PMID- 7499307 TI - Proton-translocating nicotinamide nucleotide transhydrogenase. Reconstitution of the extramembranous nucleotide-binding domains. AB - The nicotinamide nucleotide transhydrogenase of bovine mitochondria is a homodimer of monomer M(r) = 109,065. The monomer is composed of three domains, an NH2-terminal 430-residue-long hydrophilic domain I that binds NAD(H), a central 400-residue-long hydrophobic domain II that is largely membrane intercalated and carries the enzyme's proton channel, and a COOH-terminal 200-residue-long hydrophilic domain III that binds NADP(H). Domains I and III protrude into the mitochondrial matrix, where they presumably come together to form the enzyme's catalytic site. The two-subunit transhydrogenase of Escherichia coli and the three-subunit transhydrogenase of Rhodospirillum rubrum have each the same overall tridomain hydropathy profile as the bovine enzyme. Domain I of the R. rubrum enzyme (the alpha 1 subunit) is water soluble and easily removed from the chromatophore membranes. We have isolated domain I of the bovine transhydrogenase after controlled trypsinolysis of the purified enzyme and have expressed in E. coli and purified therefrom domain III of this enzyme. This paper shows that an active bidomain transhydrogenase lacking domain II can be reconstituted by the combination of purified bovine domains I plus III or R. rubrum domain I plus bovine domain III. PMID- 7499308 TI - Use of an oligoribonucleotide containing the polypurine tract sequence as a primer by HIV reverse transcriptase. AB - A primary site for initiation of plus strand DNA synthesis in human immunodeficiency virus (HIV) corresponds to a 19-nucleotide-long purine rich sequence located just upstream of the U3 region, designated the polypurine tract (PPT). The HIV reverse transcriptase (RT) uses its RNase H activity to cut the genomic RNA after minus strand DNA synthesis. A plus strand PPT primer is formed, extended, and then removed. In vitro, the HIV-RT recognizes this primer specifically, using it much more efficiently than other RNA primers. However, the PPT still primes significantly less efficiently than DNA primers. The 19 nucleotide PPT primer is partially resistant to degradation when compared with other oligoribonucleotides. Prior to initiation of DNA synthesis, several nucleotides are removed by the RT from the 3' ends of some of the PPT primers. Cleavage is enhanced in the absence of dNTPs. We suggest that DNA synthesis suppresses primer degradation, so that primer extension and cleavage occur in proper sequence. As a result of 3' end degradation, PPT elongation products contain 5'-RNA segments from 16 to 19 nucleotides in length. These shorter segments are also generated from a longer transcript containing the PPT sequence, indicating that they are not created as a result of binding of the RT to the 5' end of the PPT oligoribonucleotide. Full-length and shorter versions of the PPT primers are cleaved from the extended DNA by RT. These experiments show that HIV RT has a specificity to generate a primer in the region of the PPT but that the ends of the primer are not well defined. PMID- 7499309 TI - Unfolding properties of tryptophan-containing alpha-subunits of the Escherichia coli tryptophan synthase. AB - The urea-induced unfolding of the Escherichia coli tryptophan synthase alpha subunit is examined via fluorescence measurements with tryptophan-containing alpha-subunit mutants, constructed by in vitro mutagenesis. Early unfolding studies with urea and guanidine suggested that the wild type protein unfolded in a two-step process with a stable intermediate composed of a native alpha-1 folding unit (residues 1-188) and a completely unfolded alpha-2 folding unit (residues 189-268). Recently, more detailed spectroscopic and calorimetric data from the Matthews and Yutani groups indicate that such a structure for the intermediates seems unlikely. Previously, we described the introduction of Trp residues as unfolding reporter groups separately into each of the folding domains and showed that these proteins are wild type enzymatically and in their stability to urea. The unfolding behavior of these alpha-subunits, monitored by fluorescence intensity changes at the discrete emission lambda max for each, in both equilibrium and kinetic experiments, suggest that: (a) both folding units commence unfolding simultaneously (near 2 M urea); (b) the larger alpha-1 unit unfolds in a multistep process, initially yielding a partially unfolded intermediate form which subsequently appears to unfold progressively to completion; and (c) the smaller alpha-2 unit unfolds in a single step event. These results are also clearly incompatible with the early proposals on the structure of the intermediate. It is suggested here that the intermediate is heterogeneous, consisting of a stable, partially unfolded form of alpha-1 attached to either a completely folded or completely unfolded form of alpha-2. These results are consistent with and provide an added dimension to the recent description of the proposed structure of the intermediate. PMID- 7499311 TI - Granulocyte-colony stimulating factor and lipopolysaccharide regulate the expression of interleukin 8 receptors on polymorphonuclear leukocytes. AB - Interleukin 8 (IL-8) is a potent chemoattractant and activating factor for human polymorphonuclear leukocytes (PMN) and hence plays a critical role in the pathogenesis of acute inflammation. Two unique but homologous receptors for IL-8 have been cloned (IL-8RA and -B), each of which binds the IL-8 ligand with high affinity. PMN stimulated by cytokines or lipopolysaccharide (LPS) exhibit changes in IL-8R mRNA and 125I-IL-8 binding. Granulocyte-colony stimulating factor (G CSF) treatment of PMN enhances, and LPS inhibits, IL-8R mRNA expression. Similarly, 125I-IL-8 ligand binding to PMN is increased by G-CSF and decreased by LPS treatment. The stimulatory effect of G-CSF on IL-8R expression is transcriptional as it is inhibited by actinomycin D and is evident in nuclear run on analyses. In contrast, LPS down-regulates IL-8R by both transcriptional and post-transcriptional mechanisms. The alterations in IL-8R expression are associated with similar changes in the IL-8-induced chemotactic responses of PMN. In conclusion, the two types of IL-8 receptor differ in their cellular distribution and are regulated in response to cytokines and LPS. Regulation of IL 8R expression by endogenous and exogenous immunomodulators may be important in the in vivo control of PMN effector functions in inflammation. PMID- 7499310 TI - Thiazolidinediones and fatty acids convert myogenic cells into adipose-like cells. AB - Fatty acids and thiazolidinediones act as potent activators of the adipose differentiation program in established preadipose cell lines. In this report, the effects of these agents on the differentiation pathway of myoblasts have been investigated. Exposure of C2C12N myoblasts (a subclone of the C2C12 cell line) to thiazolidinediones or fatty acids prevents the expression of myogenin, alpha actin, and creatine kinase, thus abolishing the formation of multinucleated myotubes. These treatments lead in parallel to the expression of a typical adipose differentiation program including acquisition of adipocyte morphology and activation of adipose-related genes. A similar transition toward the adipose differentiation pathway also occurs in mouse muscle satellite cells maintained in primary culture. Thiazolidinediones exert their adipogenic effects only in non terminally differentiated myoblasts; myotubes are insensitive to the compounds. Continuous exposure to inducers after growth arrest is not required to maintain the adipose phenotype, but proliferation of adipose-like C2C12N cells leads to a complete reversion toward undifferentiated cells able to undergo either myogenic or adipogenic differentiation depending on the composition of culture medium. These results indicate that adipogenic inducers, such as thiazolidinediones or fatty acids, specifically convert the differentiation pathway of myoblasts into that of adipoblasts. PMID- 7499312 TI - Characterization of profilaggrin endoproteinase 1. A regulated cytoplasmic endoproteinase of epidermis. AB - Profilaggrin, an insoluble precursor of the intermediate filament-associated protein filaggrin, contains multiple internal repeats (PIRs). At terminal differentiation of epidermis, proteolytic processing within a "linker" region of each PIR releases soluble filaggrin in a two-stage process. The first stage endoproteinase (PEP1, profilaggrin endoproteinase 1) cleaves mouse profilaggrin at a subset of the linkers, yielding processing intermediates consisting of several filaggrin repeats. An epidermal endoproteinase that cleaves the requisite linker subset has been purified 4,966-fold from mouse epidermal extracts. SDS polyacrylamide gel electrophoresis demonstrated a band of molecular mass of 29.5 kDa that correlated with the activity. Labeling with [3H]diisopropylfluorophosphate identified PEP1 as a serine protease; inhibitor studies suggest that it is similar to chymotrypsin, as expected from previous in vivo studies. The purified PEP1 cleaved a peptide derived from profilaggrin (P1) at three residues within and adjacent to a multiple tyrosine sequence, consistent with the in vivo processing sites. No exopeptidase activity was detected. PEP1 is only active toward insoluble profilaggrin, resulting in partial solubilization, consistent with a role in dispersal of profilaggrin during terminal differentiation. In contrast to the specific cleavage of mouse profilaggrin, PEP1 cleaved all linker regions of rat profilaggrin. Studies with phosphorylated P1 suggest that PEP1 specificity may be partly regulated by profilaggrin phosphorylation. PMID- 7499313 TI - Kinetics of folding and membrane insertion of a beta-barrel membrane protein. AB - We have studied the kinetics of folding and membrane insertion of the outer membrane protein OmpA of Escherichia coli. In the native structure, its membrane inserted domain forms a beta-barrel. The protein was unfolded in solubilized form in water/urea, and refolding was induced by dilution of urea and simultaneous addition of lipid vesicles. Three transitions along the folding pathway could be distinguished. Their characteristic times lie below a second, in the range of minutes, and in the range of an hour. The fast process corresponds to the transition from the unfolded state in water/urea to a misfolded state in water, the moderately slow process to a transition from the misfolded state to a partially folded state in the membrane, and the slow process to the transition from the partially folded to the native state. The partially folded state in the membrane is interpreted as the analogue of the molten globule state of soluble proteins. PMID- 7499314 TI - Recombinant core streptavidins. A minimum-sized core streptavidin has enhanced structural stability and higher accessibility to biotinylated macromolecules. AB - Two recombinant core streptavidins were designed and characterized to understand the role of the terminal sequences, present in naturally truncated core streptavidins, in the properties of streptavidin. One recombinant core streptavidin, Stv-25, has an amino acid sequence very similar to natural core streptavidins. The other recombinant molecule, Stv-13, has further truncation of the terminal residues and consists essentially of only the beta-barrel structure characteristic of streptavidin. These recombinant core streptavidins are tetrameric and bind four biotins/molecule, as does natural streptavidin. The solubility characteristics of Stv-13, determined by varying the concentration of ammonium sulfate or ethanol, were almost the same as those of Stv-25 and natural core streptavidin. However, Stv-13 showed an enhanced structural stability compared with Stv-25 and natural core streptavidin. For example, Stv-13 retained greater than 80% of its biotin binding ability after incubation in 6 M guanidine hydrochloride at pH 1.5, under which conditions, Stv-25 and natural core streptavidin retained only about 20% of their biotin binding ability. In addition, Stv-13 showed higher accessibility to biotinylated DNA than natural core streptavidin. Apparently, the terminal regions, present on the surface of natural core streptavidin, spatially hinder biotinylated macromolecules from approaching the biotin binding sites. PMID- 7499317 TI - Glucocorticoid-induced functional polarity of growth factor responsiveness regulates tight junction dynamics in transformed mammary epithelial tumor cells. AB - The synthetic glucocorticoid, dexamethasone, induces the "normal-like" differentiated property of tight junction formation and suppresses growth of the Con8 mammary epithelial tumor cell line, derived from a 7,12 dimethylbenz(alpha)anthracene-induced rat mammary adenocarcinoma. Characterization of the transepithelial electrical resistance of Con8 mammary tumor cells cultured on permeable supports revealed that a novel response to dexamethasone is the generation of a polarized cell monolayer with respect to epidermal growth factor receptor responsiveness. Administration of transforming growth factor-alpha (TGF-alpha) to the basolateral, but not the apical, plasma membrane compartment disrupted the glucocorticoid-stimulated tight junction barrier. Confocal immunofluorescence microscopy revealed that dexamethasone caused the ZO-1 tight junction-associated protein to localize exclusively to the apical border of laterally adjacent membranes of the cell periphery, whereas basolateral administration of TGF-alpha caused the redistribution of ZO-1 back to disorganized aggregates along the cell periphery. In contrast, TGF-alpha was able to exert its mitogenic effects equally on both sides of the cell monolayer independent of its polarized disruption of tight junction formation. Our results represent the first evidence for a functional polarization of the epidermal growth factor receptor and strongly implicate the glucocorticoid-regulated formation of tight junctions in policing the polarized responsiveness of mammary cells to growth factors. PMID- 7499315 TI - Human protein disulfide isomerase functionally complements a dsbA mutation and enhances the yield of pectate lyase C in Escherichia coli. AB - Human PDI was expressed to the Escherichia coli periplasm, by using a plasmid encoded ompA-PDI fusion under the control of the trp promoter. Periplasmic extracts were shown to contain active PDI using the scrambled ribonuclease assay. PDI activity was also demonstrated by complementation of two phenotypes associated with a dsbA mutation. Alkaline phosphatase activity, which is reduced in dsbA cells, was restored to wild type levels by PDI. PelC, a pectate lyase from Erwinia carotovora, was shown to be DsbA dependent in E. coli. PDI was able to restore its activity to that seen in wild type cells. Increased expression of PDI was found to increase the yield of active PelC above that seen in wild type cells. PDI also enhanced the yield of PelC in DsbA- cells but only in the presence of exogenous oxidized glutathione. PDI is thus able to functionally substitute for DsbA in the folding of disulfide-bonded proteins in the bacterial periplasm and to enhance the yield of highly expressed protein when the ability of the E. coli periplasm to fold protein may be saturated. However, our results suggest that the activities of DsbA and PDI in vivo may be different. PMID- 7499318 TI - Free radicals generated during the glycation reaction of amino acids by methylglyoxal. A model study of protein-cross-linked free radicals. AB - The formation of alpha-dicarbonyl compounds seems to be an important step for cross-linking proteins in the glycation or Maillard reaction. To elucidate the mechanism for the cross-linking reaction, we studied the reaction between a three carbon alpha-dicarbonyl compound, methylglyoxal, and amino acids. Our results showed that this reaction generated yellow fluorescent products as formed in some glycated proteins. In addition, three types of free radical species were also produced, and their structures were determined by EPR spectroscopy. These free radicals are 1) the cross-linked radical cation, 2) the methylglyoxal radical anion as the counterion, and 3) the superoxide radical anion produced only in the presence of oxygen. The generation of the cross-linked radical cations and the methylglyoxal radical anions does not require metal ions or oxygens. These results indicate that dicarbonyl compounds cross-link free amino groups of protein by forming Schiff bases, which donate electrons directly to dicarbonyl compounds to form the cross-linked radical cations and the methylglyoxal radical anions. Oxygen can accept an electron from the radical anion to generate a superoxide radical anion, which can initiate damaging chain reactions. Time course studies suggest that the cross-linked radical cation is a precursor of yellow fluorescent glycation end products. PMID- 7499316 TI - Cloning of a novel RNA binding polypeptide (RA301) induced by hypoxia/reoxygenation. AB - Astrocytes have a critical role in the neuronal response to ischemia, as their production of neurotrophic mediators can favorably impact on the extreme sensitivity of nervous tissue to oxygen deprivation. Using a differential display method, a novel putative RNA binding protein, RA301, was cloned from reoxygenated astrocytes. Analysis of the deduced amino acid sequence showed two ribonucleoprotein domains and serine/arginine-rich domains, suggestive of their function as RNA splicing factor. Northern analysis displayed striking induction only in cultured astrocytes within 15 min of reoxygenation and reached a maximum by 60 min after hypoxia/reoxygenation. Immunoblotting demonstrated expression of an immunoreactive polypeptide of the expected molecular mass, 36 kDa, in lysates of hypoxia/reoxygenated astrocytes. Induction of RA301 mRNA was mediated, in large part, by endogenously generated reactive oxygen species, as shown by diphenyl iodonium, an inhibitor of neutrophil-type nicotinamide adenine dinucleotide phosphate oxidase which blocks oxygen-free radical formation by astrocytes. Similarly, increased expression of RA301 in supporting a neurotrophic function of astrocytes was suggested by inhibition of interleukin-6 elaboration, a neuroprotective cytokine, in the presence of antisense oligonucleotide for RA301. These studies provide a first step in characterizing a novel putative RNA binding protein, whose expression is induced by oxygen-free radicals generated during hypoxia/reoxygenation, and which may have an important role in redirection of biosynthetic events observed in the ischemic tissues. PMID- 7499319 TI - Molecular and structural characterization of the heat-resistant thyroxine-binding globulin-Chicago. AB - Thyroxine-binding globulin (TBG) is the main transport protein for thyroxine (T4) in blood. It shares considerable sequence homology with alpha 1-antitrypsin (AT) and other members of the serine proteinase inhibitor (serpin) superfamily of proteins. The crystallographic structure of AT has been determined and was found to represent the archetype of the serpins. This model has been used for structure function correlations of TBG. Sequence analysis of the heat-resistant variant TBG Chicago (TBG-CH) revealed a substitution of the normal tyrosine 309 with phenylalanine. For further analysis, vectors containing the coding regions of normal TBG (TBG-N) and TBG-CH were constructed, transcribed in vitro, and expressed in Xenopus oocytes. Both TBGs were secreted into the culture medium and could not be distinguished by gel electrophoresis. Scatchard analysis of T4 binding to TBG-N and -CH revealed no significant differences in binding affinity. The rate of heat denaturation of TBGs was determined by measurement of residual T4 binding capacity after incubation at 60 degrees C for various periods of time. The half-life values of denaturation of TBG-N and -CH were 7 and 132 min, respectively. The tyrosine 309 to phenylalanine substitution of TBG-CH involves a highly conserved phenylalanine residue of the serpins. The respective phenylalanine 312 of AT ties the alpha-helix hI1 to the molecule, thus stabilizing the tertiary structure. A substitution with tyrosine would disrupt this interaction. Accordingly, stabilization of the TBG molecule by replacement of tyrosine with phenylalanine in position 309 causes the increased heat stability of TBG-CH. PMID- 7499321 TI - Characterization of a phosphoprotein phosphatase for the phosphorylated form of nucleoside-diphosphate kinase from Pseudomonas aeruginosa. AB - Nucleoside-diphosphate kinase (ATP:nucleoside-diphosphate phosphotransferase, EC 2.7.4.6; NDP kinase) is an important enzyme for the maintenance of the correct cellular levels of nucleoside triphosphates (NTPs) and their deoxy derivatives (dNTPs) and is involved in the regulation of cellular development. The enzyme is under the dual control of algR2 and algH in Pseudomonas aeruginosa. We report here the purification and characterization of a protein that dephosphorylates the phosphorylated intermediate form of the P. aeruginosa NDP kinase (Ndk). Dephosphorylation of Ndk phosphate leads to loss of its enzymatic activity. The 10.1-kDa polypeptide shows 77% homology at the N terminus with the Spo0E phosphatase, identified as a negative regulator of sporulation in Bacillus subtilis and 66% with the human Bax protein, identified as an effector of programmed cell death. The phosphatase termed Npp showed varied specificity toward phosphorylated Ndks from different sources including human erythrocyte Ndk phosphate. Its activity toward other histidine phosphates such as CheA or the alpha-subunit of succinyl-CoA synthetase or phosphoesters such as p-nitrophenyl phosphate was quite limited. Npp was stable at room temperature up to 2 h and required Mg2+ for activity. The presence of a phosphatase capable of dephosphorylating the phosphorylated form of P. aeruginosa Ndk represents an interesting and efficient mode of post-translational modification of an enzyme crucial to cellular development. PMID- 7499320 TI - How do volatile anesthetics inhibit Ca(2+)-ATPases? AB - Volatile anesthetics at concentrations that are used in clinical practice to induce anesthesia selectively inhibit activity of the plasma membrane Ca(2+) transport ATPase (Kosk-Kosicka, D., and Roszczynska, G. (1993) Anesthesiology 79, 774-780). We have investigated the mechanism of the inhibitory action of several anesthetics on the purified erythrocyte Ca(2+)-ATPase by employing fluorescence spectroscopy measurements that report changes in the environment of intrinsic tryptophans and of an extrinsic probe attached in the active site of the enzyme. We have shown that the observed inhibition of the Ca(2+)-dependent activation of the enzyme correlates well with the elimination of the Ca(2+)-induced conformation change that is important for the proper function of the enzyme. Analysis of the anesthetics effects on the total tryptophan fluorescence indicates a significant effect on enzyme conformation. Similar changes have been observed in the sarcoplasmic reticulum Ca(2+)-ATPase. We propose that volatile anesthetics inhibit Ca(2+)-ATPase by interacting with nonpolar sites in protein interior, in analogy to the binding demonstrated for myoglobin, hemoglobin, and adenylate kinase (Schoenborn, B. P., and Featherstone, R. M. (1967) Adv. Pharmacol. 5, 1-17; Tilton, R. F., Kuntz, I. D., and Petsko, G. A. (1984) Biochemistry 23, 2849-2857). Such binding is expected to modify conformational substate(s) of the enzyme and perturb its function. We view this process as an example of a general phenomena of interaction of small molecules with internal sites in proteins. PMID- 7499322 TI - The spatio-temporal control of the expression of glutamine synthetase in the liver is mediated by its 5'-enhancer. AB - In previous studies of the glutamine synthetase gene, the promoter and two enhancer elements, one in the upstream region and one within the first intron, were identified. To analyze the role of the far-upstream enhancer element in the regulation of the expression of the glutamine synthetase gene, two classes of transgenic mice were generated. In GSK mice, the basal promoter directs the expression of the chloramphenicol acetyltransferase reporter gene. In GSL mice reporter gene expression is driven, in addition, by the upstream regulatory region, including the far-upstream enhancer. Whereas chloramphenicol acetyltransferase expression was barely detectable in GSK mice, high levels were detected in GSL mice. By comparing chloramphenicol acetyltransferase expression with that of endogenous glutamine synthetase in GSL mice, three groups of organs were distinguished in which the effects of the upstream regulatory region on the expression of glutamine synthetase were quantitatively different. The chloramphenicol acetyltransferase mRNA in the GSL mice was shown to be localized in the pericentral hepatocytes of the liver. The developmental changes in chloramphenicol acetyltransferase enzyme activity in the liver were similar to those in endogenous glutamine synthetase. These results show that the upstream region is a major determinant for three characteristics of glutamine synthetase expression: its organ specificity, its pericentral expression pattern in the liver, and its developmental appearance in the liver. PMID- 7499323 TI - Levels and alternative splicing of amyloid beta protein precursor (APP) transcripts in brains of APP transgenic mice and humans with Alzheimer's disease. AB - Abnormal expression of human amyloid precursor protein (hAPP) gene products may play a critical role in Alzheimer's disease (AD). Recently, a transgenic model was established in which platelet-derived growth factor (PDGF) promoter-driven neuronal expression of an alternatively spliced hAPP minigene resulted in prominent AD-type neuropathology (Games, D., Adams, D., Alessandrini, R., Barbour, R., Berthelette, P., Blackwell, C., Carr, T., Clemens, J., Donaldson, T., Gillespie, F., Guido, T., Hagopian, S., Johnson-Wood, K., Khan, K., Lee, M., Leibowitz, P., Lieberburg, I., Little, S., Masliah, E., McConlogue, L., Montoya Zavala, M., Mucke, L., Paganini, L., and Penniman, E. (1995) Nature 373, 523 527). Here we compared the levels and alternative splicing of APP transcripts in brain tissue of hAPP transgenic and nontransgenic mice and of humans with and without AD. PDGF-hAPP mice showed severalfold higher levels of total APP mRNA than did nontransgenic mice or humans, whereas their endogenous mouse APP mRNA levels were decreased. This resulted in a high ratio of mRNAs encoding mutated hAPP versus wild-type mouse APP. Modifications of hAPP introns 6, 7, and 8 in the PDGF-hAPP construct resulted in a prominent change in alternative splice site selection with transcripts encoding hAPP770 or hAPP751 being expressed at substantially higher levels than hAPP695 mRNA. Frontal cortex of humans with AD showed a subtle increase in the relative abundance of hAPP751 mRNA compared with normal controls. These data identify specific intron sequences that may contribute to the normal neuronspecific alternative splicing of APP pre-mRNA in vivo and support a causal role of hAPP gene products in the development of AD type brain alterations. PMID- 7499324 TI - Fluorescent labeling of purified beta 2 adrenergic receptor. Evidence for ligand specific conformational changes. AB - The purpose of the present study was to develop an approach to directly monitor structural changes in a G protein-coupled receptor in response to drug binding. Purified human beta 2 adrenergic receptor was covalently labeled with the cysteine-reactive, fluorescent probe N,N'-dimethyl-N-(iodoacetyl)-N'-(7-nitrobenz 2-oxa-1,3-diazol-4- yl)ethylenediamine (IANBD). IANBD is characterized by a fluorescence which is highly sensitive to the polarity of its environment. We found that the full agonist, isoproterenol, elicited a stereoselective and dose dependent decrease in fluorescence from IANBD-labeled beta 2 receptor. The change in fluorescence could be plotted against the concentration of isoproterenol as a simple hyperbolic binding isotherm demonstrating interaction with a single binding site in the receptor. The ability of several adrenergic antagonists to reverse the response confirmed that this binding site is identical to the well described binding site in the beta 2 receptor. Comparison of the response to isoproterenol with a series of adrenergic agonists, having different biological efficacies, revealed a linear correlation between biological efficacy and the change in fluorescence. This suggests that the agonist-mediated decrease in fluorescence from IANBD-labeled beta 2 receptor is due to the same conformational change as involved in receptor activation and G protein coupling. In contrast to agonists, negative antagonists induced a small but significant increase in base line fluorescence. Despite the small amplitude of this response, it supports the notion that antagonists by themselves may alter receptor structure. In conclusion, our data provide the first direct evidence for ligand-specific conformational changes occurring in a G protein-coupled receptor. Furthermore, the data demonstrate the potential of fluorescence spectroscopy as a tool for further delineating the molecular mechanisms of drug action at G protein-coupled receptors. PMID- 7499325 TI - Molecular cloning of an intracellular P-type ATPase from Dictyostelium that is up regulated in calcium-adapted cells. AB - Results from a number of laboratories suggest that intracellular Ca2+ is involved in the regulation of Dictyostelium discoideum growth and development. To learn more about the regulation and function of intracellular Ca2+ in this organism, we have cloned and sequenced cDNAs that encode a putative P-type Ca2+ ATPase designated patA. The deduced protein product of this gene (PAT1) has a calculated molecular mass of 120,718 daltons. It exhibits about 46% amino acid identity with Ca2+ ATPases of the plasma membrane Ca2+ ATPase family and lower identity with sarco(endo)plasmic reticulum Ca2+ ATPase family members and monovalent cation pumps. However, PAT1 lacks the highly conserved calmodulin-binding domain present in the C-terminal region of most plasma membrane Ca2+ ATPase-type enzymes. When Dictyostelium amoebae are adapted to grow in the presence of 80 mM CaCl2, both the patA message and protein product are up-regulated substantially. These cells also exhibit an increase in the rate and magnitude of intracellular P-type Ca2+ uptake activity. Immunofluorescence analysis indicates that PAT1 colocalizes with bound calmodulin to intracellular membranes, probably components of the contractile vacuole complex. The presence of PAT1 on the contractile vacuole suggests that in Dictyostelium this organelle might function in Ca2+ homeostasis as well as in water regulation. PMID- 7499326 TI - Membrane topology analysis of the sensor kinase KdpD of Escherichia coli. AB - The expression of the kdpFABC operon, coding for the K(+)-translocating Kdp ATPase, is under the control of the two regulatory proteins KdpD and KdpE, which belong to the group of sensor kinase/response regulator systems. The topology of the KdpD protein in the cytoplasmic membrane was investigated using LacZ and PhoA fusions at different sites within the polypeptide chain and by treating spheroplasts in the presence or absence of Triton X-100 with the protease kallikrein. The results revealed that KdpD has four membrane-spanning segments in the middle of the polypeptide chain, whereas N and C terminus are both cytoplasmic. PMID- 7499327 TI - Heparin modulates the binding of insulin-like growth factor (IGF) binding protein 5 to a membrane protein in osteoblastic cells. AB - Osteoblast-like cells secrete insulin-like growth factor (IGF) binding protein-5 (IGFBP-5), which may act to enhance IGF-stimulated osteoblast function. We recently demonstrated that carboxyl-truncated IGFBP-5 (IGFBP-5(1-169)) binds to the osteoblast surface and stimulates mitogenesis by a pathway that is independent of IGF action. The present study was conducted to determine the mechanism of osteoblast binding of IGFBP-5, beginning with the assumption that cell surface glycosaminoglycans may mediate the binding of this heparin binding protein. Intact 125I-IGFBP-5 and 125I-IGFBP-5(1-169) exhibited one-site binding to mouse osteoblast monolayers with dissociation constants of 28 and 6 nM for intact 125I-IGFBP-5 and 125I-IGFBP-5(1-169), respectively. Osteoblast binding of intact 125I-IGFBP-5 was inhibited by low heparin concentrations, while 125I-IGFBP 5(1-169) binding was stimulated by heparin. Treatment of cells with heparinase or chlorate to decrease surface glycosaminoglycan density failed to reduce the binding of either form of IGFBP-5. In contrast, pretreatment of cells with IGFBP 5 caused down-regulation of 125I-IGFBP-5 binding. Cross-linking studies revealed that both intact 125I-IGFBP-5 and 125I-IGFBP-5(1-169) bind to proteins in Triton extracts of osteoblast membranes, which were absent in osteoblast-derived matrix. Purification of membrane extracts by IGFBP-5 affinity chromatography revealed a 420-kDa band on reduced SDS-polyacrylamide gels. While the membrane protein internalized both forms of IGFBP-5, heparin treatment inhibited the internalization of intact 125I-IGFBP-5 but stimulated 125I-IGFBP-5(1-169) internalization. These data indicate that IGFBP-5 binds to and is internalized by an osteoblast membrane protein, which does not appear to be a proteoglycan. Glycosaminoglycans, however, modulate the binding and internalization of IGFBP-5 in a way that may preferentially favor the intracellular accumulation of the carboxyl-truncated form. PMID- 7499328 TI - Sequence-specific RNA recognition by the Xenopus Y-box proteins. An essential role for the cold shock domain. AB - The Xenopus Y-box protein FRGY2 has a role in the translational silencing of masked maternal mRNA. Here, we determine that FRGY2 will recognize specific RNA sequences. The evolutionarily conserved nucleic acid-binding cold shock domain is required for sequence-specific interactions with RNA. However, RNA binding by FRGY2 is facilitated by N- and C-terminal regions flanking the cold shock domain. The hydrophilic C-terminal tail domain of FRGY2 interacts with RNA independent of the cold shock domain but does not determine sequence specificity. Thus, both sequence-specific and nonspecific RNA recognition domains are contained within the FRGY2 protein. PMID- 7499329 TI - Growth inhibition of hepatoma cells induced by vitamin K and its analogs. AB - Congeners of vitamin K are known to inhibit cell growth, although the precise mechanisms of growth inhibition are not well understood. To investigate the mechanisms involved, we synthesized several vitamin K analogs and examined their growth inhibitory activities for a human hepatoma cell line (Hep3B). The analogs included 2-methyl-1,4-naphthoquinone and trimethyl-benzoquinone, with and without aliphatic side chains at position 3. The side chains were all-carbon, thioethers, or O-ethers. Growth inhibition was potent in the compounds with short chains. The presence of a sulfur (thioether) or oxygen atom (O-ether) at the site of attachment of the side chain to the ring potentiated the activity. Apoptotic cell death was induced by the potent growth inhibitory compounds at low concentrations (20-60 microM), whereas necrotic cell death followed treatment with the same compounds at high concentrations. Expression of c-myc, which is thought to be associated with apoptosis, was increased by most of the compounds tested. Both reduced glutathione and cysteine almost completely abrogated the growth inhibitory effects of the thioether analogs as well as of vitamin K3. The effect of glutathione was less prominent for the all-carbon and O-ether analogs, and cysteine had no effect on these analogs. Catalase and deferoxamine mesylate had no significant effect on the thioether analogs, although they showed partial antagonistic effects on the growth inhibition of vitamin K3 and the all-carbon and O-ether analogs. Other non-thiol antioxidants tested had no effect on any of the analogs. Our results indicated that vitamin K-related quinoid compounds cause growth inhibition and both apoptotic and necrotic cell death and that the effects may be mediated by interaction at position 3 of their quinoid nuclei with cellular thiols. PMID- 7499330 TI - Transfection of N-acetylglucosaminyltransferase III gene suppresses expression of hepatitis B virus in a human hepatoma cell line, HB611. AB - beta-D-mannoside beta-1,4-N-acetylglucosaminyltransferase III (GnT-III) catalyzes the addition of N-acetylglucosamine in beta 1-4 linkage to the beta-linked mannose of the trimannosyl core of N-linked oligosaccharides and forms a bisecting GlcNAc structure. Although the biological meaning of the bisecting GlcNAc structure remains unclear, it is known that the attachment of a bisecting GlcNAc inhibits further processing of oligosaccharides by other glycosyltransferases. To investigate whether or not structural changes of oligosaccharides affect secretion and gene expression of hepatitis B virus (HBV), we introduced the GnT-III gene into a human hepatoma cell line, HB611, which secreted HBV-related proteins into the medium. Positive transfectants were cloned by hygromycin resistant selection. Three clones have high activities of GnT-III and secreted lower levels of HBV-related proteins into the medium in comparison with other clones. These clones showed marked suppression of HBV-related mRNAs and an increased binding with E-PHA as judged by lectin blot. Expression of beta actin, alpha fetoprotein, albumin, and prealubmin was not correlated with GnT-III activity in all the seven clones. Treatment of these cells with tunicamycin or swainsonine resulted in enhanced expression of HBV-related mRNA. These results indicate that some glycoproteins whose oligosaccharide structures are changed by over-expression of GnT-III suppress HBV gene expression. PMID- 7499332 TI - Differential activation of mitogen-activated protein kinase and S6 kinase signaling pathways by 12-O-tetradecanoylphorbol-13-acetate (TPA) and insulin. Evidence for involvement of a TPA-stimulated protein-tyrosine kinase. AB - AG-18, an inhibitor of protein-tyrosine kinases, was employed to study the role of tyrosine-phosphorylated proteins in insulin- and phorbol ester-induced signaling cascades. When incubated with Chinese hamster ovary cells overexpressing the insulin receptor, AG-18 reversibly inhibited insulin-induced tyrosine phosphorylation of insulin receptor substate-1, with minimal effects either on receptor autophosphorylation or on phosphorylation of Shc64. Under these conditions, AG-18 inhibited insulin-stimulated phosphorylation of the ribosomal protein S6, while no inhibition of insulin-induced activation of mitogen-activated protein kinase (MAPK) kinase or MAPK was detected. In contrast, 12-O-tetradecanoylphorbol-13-acetate (TPA)-induced activation of MAPK kinase and MAPK and phosphorylation of S6 were inhibited by AG-18. This correlated with inhibition of TPA-stimulated tyrosine phosphorylation of several proteins, the most prominent ones being pp114 and pp120. We conclude that Tyr-phosphorylated insulin receptor substrate-1 is the main upstream regulator of insulin-induced S6 phosphorylation by p70s6k, whereas MAPK signaling seems to be activated in these cells primarily through the adaptor molecule Shc. In contrast, TPA-induced S6 phosphorylation is mediated by the MAPK/p90rsk cascade. A key element of this TPA stimulated signaling pathway is an AG-18-sensitive protein-tyrosine kinase. PMID- 7499331 TI - Possible participation of autocrine and paracrine vascular endothelial growth factors in hypoxia-induced proliferation of endothelial cells and pericytes. AB - Hypoxia is the principal factor that causes angiogenesis. These experiments were conducted to explore how it induces the proliferation of vascular cells, a key step in angiogenesis. Human umbilical vein endothelial cells and bovine retinal pericytes were grown in controlled atmosphere culture chambers containing various concentrations of oxygen. The numbers of both endothelial cells and pericytes increased significantly under hypoxic conditions; the O2 concentrations that achieved maximal growth promotion were 10% for endothelial cells and 2.5% for pericytes. Quantitative reverse transcription-polymerase chain reaction analysis revealed that mRNAs coding for the secretory forms of vascular endothelial growth factor (VEGF), a mitogen specific to endothelial cells, were present in both endothelial cells and pericytes and that their levels increased significantly in the two cell types as the atmospheric O2 concentration decreased. The two genes for VEGF receptors, kinase insert domain-containing receptor (kdr) and fms-like tyrosine kinase 1 (flt1), were found to be constitutively expressed in endothelial cells, and their relative mRNA levels were ranked in that order. On the other hand, only flt1 mRNA was detected in pericytes under hypoxic conditions. Furthermore, most antisense oligodeoxyribonucleotides complementary to VEGF mRNAs efficiently inhibited DNA synthesis in endothelial cells cultured under hypoxic conditions. These results indicate that autocrine and paracrine VEGFs may take part in the hypoxia-induced proliferation of endothelial cells. PMID- 7499333 TI - Lysine residues at positions 234 and 236 in yeast porin are involved in its assembly into the mitochondrial outer membrane. AB - Various point mutations of lysyl residues in yeast mitochondrial porin (283 residues) were tested for their ability to assemble in vitro into the outer membranes of intact yeast mitochondria. Assembly was evaluated by protection from proteinases. The extent of assembly of two of the mutants, K234E and K236E porins, was much less than for wild-type in either post-translational or co translational assembly assays. Lysine to glutamate mutants at other positions and K234R porin assembled as well as wild-type, but K234Q porin was poorly inserted. When both Lys-234 and Lys-236 were mutated, K234R/K236R porin was inserted better than K234Q/K236Q porin, which was inserted better than K234E/K236E; however, none of these mutants assembled as well as wild-type porin. It was concluded that optimal assembly of yeast porin depended on the presence of positively charged residues at both positions 234 and 236 and a lysine at one of these positions. After undergoing the assembly reaction, mutants that were vulnerable to proteinase K (i.e. K234E, K234Q, and K236E porins) seemed to be incompletely digested and were, to varying degrees, resistant to extraction by Na2CO3 (pH 11.5). These experiments suggested that these mutants were incompletely inserted into the outer membrane. Both Lys-234 and Lys-236 are included in an internal pentapeptide, VKAKV, that is conserved in porins from protists, plants, and animals, and it is possible that, at least, the lysines in this tract are one of the signals for the membrane assembly of these proteins. PMID- 7499334 TI - Gastrin and glycine-extended progastrin processing intermediates induce different programs of early gene activation. AB - We recently reported that gastrin and glycine-extended progastrin processing intermediates (G-Gly) exert growth-promoting effects on AR4-2J cells (derived from rat pancreas) via interaction with distinct receptors. In this study we sought to investigate the mechanisms by which gastrin and G-Gly stimulate cell proliferation. While gastrin increased [Ca2+]i in AR4-2J cells, G-Gly had no effect. Similarly, G-Gly had no effect either on basal and 10(-7) M vasoactive intestinal polypeptide-stimulated cAMP generation, although gastrin is known to inhibit cAMP generation. Gastrin dose dependently stimulated AR4-2J cell mRNA content of both c-fos and c-jun, two genes known to function in regulating cell proliferation, but G-Gly had no effect. Gastrin also induced the expression of luciferase in AR4-2J cells transfected with a construct consisting of a luciferase reporter gene coupled to the serum response element of the c-fos gene promoter. In similar fashion, gastrin stimulated the activity of mitogen activated protein kinase, an enzyme known to mediate the induction of the c-fos serum response element in response to growth factor stimulation. Although G-Gly had none of these effects of gastrin in AR4-2J cells, it stimulated activity of c Jun amino-terminal kinase, an enzyme known to phosphorylate and transcriptionally activate c-Jun. These data support the notion that gastrin stimulates cell proliferation by inducing c-fos and c-jun gene expression, while G-Gly acts by post-translationally regulating early gene transcriptional activation. Our studies represent a novel model in which both the precursor and the product of a key processing reaction, peptide alpha-amidation, act cooperatively to stimulate cell proliferation via distinct receptors linked to different signal transduction pathways. PMID- 7499336 TI - Characterization of the 5'-flanking region of the gene for the gamma chain of human fibrinogen. AB - The 5'-flanking region of the gene coding for the gamma chain of human fibrinogen was characterized for its promoter activity. Reporter gene studies using chloramphenicol acetyltransferase as the indicator along with mutations in the DNA showed that a TATA-like sequence (-20 to -23 base pairs (bp)), a CAAT-like sequence (-54 to -57 bp), and an upstream stimulatory factor (USF) binding site ( 66 to -77 bp) constitute a minimal promoter that mediates liver-specific expression of the gene. Electrophoretic gel mobility assays and antibody binding studies confirmed the interaction of USF with the binding site. An IL-6 responsive element with a sequence of CTGGAA located at -301 to -306 bp was shown to be a functional element in the IL-6 response. In contrast to the IL-6 responsive elements in the human alpha- and beta-fibrinogen genes, the element in the gene for the gamma chain of human fibrinogen was unaffected by the presence or elevated levels of the beta or delta isoforms of the CCAAT/enhancer binding proteins. A negative element with sequence homology to several silencer elements was also identified in the region of -348 to -390 bp of the gene for the gamma chain of human fibrinogen. A comparison of the regulatory elements in the genes coding for all three chains in fibrinogen is also presented. PMID- 7499335 TI - Characterization of the 5'-flanking region of the gene for the alpha chain of human fibrinogen. AB - The 5'-flanking region of the gene coding for the alpha chain of human fibrinogen was isolated, sequenced, and characterized. The principal site of transcription initiation was determined by primer extension analysis and the RNase protection assay and shown to be at an adenine residue located 55 nucleotides upstream from the initiator methionine codon, or 13,399 nucleotides down-stream from the polyadenylation site of the gene coding for the gamma chain. Transient expression of constructs containing sequentially deleted 5'-flanking sequences of the alpha chain gene fused to the chloramphenicol acetyltransferase reporter gene showed that the promoter was liver-specific and inducible by interleukin 6 (IL-6). The shortest DNA fragment with significant promoter activity and full response to IL 6 stimulation encompassed the region from -217 to +1 base pairs (bp). Although six potential IL-6 responsive sequences homologous to the type II IL-6 responsive element were present, a single sequence of CTGGGA localized from -122 to -127 bp was shown to be a functional element in IL-6 induction. A hepatocyte nuclear factor 1 (HNF-1) binding site, present from -47 to -59 bp, in combination with other upstream elements, was essential for liver-specific expression of the gene. A functional CCAAT/enhancer binding protein site (C/EBP, -134 to -142 bp) was also identified within 217 bp from the transcription initiation site. An additional positive element (-1393 to -1133 bp) and a negative element (-1133 to 749 bp) were also found in the upstream region of the alpha-fibrinogen gene. PMID- 7499337 TI - Cell cycle phase-specific phosphorylation of human topoisomerase II alpha. Evidence of a role for protein kinase C. AB - Type II topoisomerases are essential for faithful cell division in all organisms. In human cells, the alpha isozyme of topoisomerase II has been implicated in catalyzing mitotic chromosome segregation via its action as a DNA unlinking enzyme. Here, we have shown that the enzymatic activity of topoisomerase II alpha protein purified from HeLa cell nuclei was strongly enhanced following phosphorylation by protein kinase C. We have investigated the possibility that this kinase is involved in cell cycle phase-specific phosphorylation of topoisomerase II alpha in HeLa cells. Two-dimensional tryptic phosphopeptide mapping revealed that topoisomerase II alpha protein immunoprecipitated from metabolically labeled HeLa cells was differentially phosphorylated during the G2/M phases of the cell cycle. To identify sites of phosphorylation, and the kinase(s) responsible for this modification, oligohistidine-tagged recombinant domains of topoisomerase II alpha protein were overexpressed in Escherichia coli and purified by affinity chromatography. Phosphorylation of a short fragment of the N-terminal ATPase domain of topoisomerase II alpha by protein kinase C in vitro generated two phosphopeptides that co-migrated with prominent G2/M phase specific phosphopeptides from the HeLa cell-derived topoisomerase II alpha protein. Site-directed mutagenesis studies indicated that phosphorylation of serine 29 generated both of these phosphopeptides. Our results implicate protein kinase C in the cell cycle phase-dependent modulation of topoisomerase II alpha enzymatic activity in human cells. PMID- 7499338 TI - The mouse bone morphogenetic protein-4 gene. Analysis of promoter utilization in fetal rat calvarial osteoblasts and regulation by COUP-TFI orphan receptor. AB - Bone morphogenetic protein-4 (BMP-4) is one of a member of related polypeptides that are important in bone formation and other developmental processes. We isolated the BMP-4 gene from a mouse genomic library and characterized the exon intron structure and one of the candidate promoters. Two alternative 5'-noncoding exons, 1A and 1B, were identified by reverse transcription polymerase chain reaction assays. Quantitative competitive polymerase chain reaction using Exon 1A, Exon 1B, and Exon 3 primers indicate the 1A-containing transcript is the primary BMP-4 mRNA expressed in bone cell cultures. Primer extension analysis supports that 1A is the major promoter utilized in bone cell cultures as well as in 9.5-day mouse embryos. 1A promoter activity indicate selective DNA regions functional in bone cells. We found potential regulatory response regions in the 1A 5'-flanking region of the BMP-4 gene for the chicken ovalbumin upstream transcription factor I (COUP-TFI). Specific binding to the COUP-TFI response regions in the BMP-4 1A promoter was demonstrated. By co-transfection of a COUP TFI expression plasmid with the BMP-4 1A promoter in fetal rat calvarial osteoblasts, we demonstrated that COUP-TFI inhibits the BMP-4 promoter activity. This suggests that COUP-TFI could act as a silencer for BMP-4 transcription in vivo. PMID- 7499339 TI - The traY gene product and integration host factor stimulate Escherichia coli DNA helicase I-catalyzed nicking at the F plasmid oriT. AB - F plasmid conjugative transfer is initiated by the introduction of a site- and strand-specific nick within the plasmid origin of transfer (oriT). Genetic studies have shown nick formation to be dependent on both the traI and traY genes. However, highly purified TraIp, the traI gene product, nicks oriT in a site- and strand-specific manner in the absence of the traY gene product (TraYp) in vitro (Matson, S.W., and Morton, B.S. (1991) J. Biol. Chem. 266, 16232-16237). Analysis of the oriT region has revealed binding sites for TraYp and the host protein integration host factor (IHF). To explore possible interactions occurring at oriT, highly purified TraIp, TraYp, and IHF were incubated with a supercoiled oriT-containing DNA substrate. A marked enhancement of the nicking reaction catalyzed by TraIp was observed in a reaction that required both TraYp and IHF. In addition, TraIp was able to nick a linear oriT-containing double-stranded DNA substrate when IHF and TraYp were present in the reaction; such a substrate is not nicked by TraIp alone. Individual protein concentration requirements for the supercoiled and linear nicking reactions were similar, and the reactions occurred at equal velocity, suggesting that they are biochemically identical. Concentrations of TraYp and IHF that yield half-maximal activity in the nicking assays compare well with the reported KD values for the IHF and TraYp binding sites in oriT. These data, coupled with data presented in the accompanying report, suggest that TraYp and IHF bind independent of one another, forming a nucleo-protein complex with oriT that can be recognized and nicked by TraIp. PMID- 7499340 TI - Stepwise assembly of a relaxosome at the F plasmid origin of transfer. AB - A central step in the transfer of genetic information during bacterial conjugation of the Escherichia coli F plasmid involves the formation of a protein DNA complex, called the relaxosome, at the origin of transfer. During conjugation, the relaxosome introduces a site- and strand-specific nick from which the physical transfer of a single strand of DNA is initiated. At least two F-encoded proteins, TraIp (traI gene product) and TraYp (traY gene product), and one host-encoded protein, integration host factor, are involved in this process. In this report, we use DNase I protection and electron microscopic techniques to investigate the mechanism of relaxosome formation. Our results show that TraYp and integration host factor form a protein-DNA complex that facilitates the binding of TraIp to assemble a relaxosome capable of introducing a site- and strand-specific nick at the origin of transfer. This nick is identical to that observed during conjugation. PMID- 7499341 TI - Increased efficiency of GroE-assisted protein folding by manganese ions. AB - This study addresses the role of ATP-bound and free Mg2+ and Mn2+ ions in the activation and modulation of chaperonin-assisted refolding of urea-denatured malate dehydrogenase. As compared with Mg2+, Mn2+ ions caused a significant increase in the rate of GroE-assisted malate dehydrogenase refolding and, concomitantly, a decrease in the rate of ATP hydrolysis. Moreover, Mn2+ increases the affinity of GroES for GroEL, even in the presence of saturating amounts of Mg2+. Chemical cross-linking showed that lower concentrations of Mn-ATP as compared with Mg-ATP are needed to form both asymmetric GroEL14GroES7 and symmetric GroEL14(GroES7)2 particles. The manganese-dependent increase in the rate of protein folding concurred with a specific increase in the amount of symmetric GroEL14-(GroES7)2 particles detected in a chaperonin solution. Thus, Mn2+ is a cofactor that can markedly increase the efficiency of the chaperonin reaction in vitro. Mn2+ ions can serve as an important tool for analyzing the molecular mechanism and the structure of chaperonins. PMID- 7499342 TI - An iron-regulated gene, magA, encoding an iron transport protein of Magnetospirillum sp. strain AMB-1. AB - Magnetospirillum sp. AMB-1 is a freshwater magnetic bacterium which synthesizes intracellular particles of magnetite (Fe3O4). A genomic DNA fragment required for synthesis of magnetic particles was previously isolated from a nonmagnetic transposon Tn5 mutant. We have determined the complete nucleotide sequence of this fragment. The 2975-base pair region contains two putative open reading frames. One open reading frame, designated magA, encodes a polypeptide which is homologous to the cation efflux proteins, the Escherichia coli potassium ion translocating protein, KefC, and the putative Na+/H(+)-antiporter, NapA, from Enterococcus hirae. Northern hybridization demonstrated that the magA mRNA transcript is 1.3 kilobases in size, corresponding to the size of the magA gene. A functional promoter was located upstream from the magA gene, and the transcription in AMB-1 was regulated by environmental iron concentration. Vesicles isolated from E. coli in which the MagA protein was expressed exhibited iron accumulation ability. We consider that the MagA protein is an iron transport involved in the synthesis of magnetic particles in AMB-1. PMID- 7499344 TI - Inhibition of gene expression by triple helix formation in hepatoma cells. AB - The aim of this study was to selectively inhibit human mitochondrial aldehyde dehydrogenase (ALDH2) gene expression by triple helix assembly. Eight 21-mer oligodeoxyribonucleotides were designed to bind to two purine-rich sequences in the 5'-flanking region of the human ALDH2 gene. Gel mobility shift assays showed that triplex formation is sequence-specific for the target duplex and the third strand oligonucleotide. In the presence of Mg2+, but absence of K+, triplex forming oligonucleotides bind to their target sites with apparent dissociation constants (Kd) in the 10(-7) to 10(-9) M range. Potassium cation virtually suppressed the triplex formation of G-C-rich duplex DNA with natural oligonucleotides, but did not prevent triplex formation with phosphorothioate modified oligonucleotides. Phosphorothioate-modified oligonucleotides were delivered into human hepatoma Hep G2 cells by cationic liposomes. The reduction in ALDH2 mRNA levels in the cells was determined by the competitive reverse transcription-polymerase chain reaction. One of the phosphorothioate-modified oligonucleotides designed to forma an antiparallel triplex with a target in the 5'-flanking region of human ALDH2 gene (-105 to -125 from the translation initiation codon ATG) reduced by 80-90% the ALDH2 mRNA levels without affecting albumin mRNA levels. Data suggest that triple-helix formation may provide a means to selectively inhibit hepatic ALDH2 gene expression for therapeutic use. PMID- 7499343 TI - Localization of furin to the trans-Golgi network and recycling from the cell surface involves Ser and Tyr residues within the cytoplasmic domain. AB - Furin is a membrane-associated endoprotease that catalyzes cleavage of precursor proteins at Arg-X-Lys/Arg-Arg sites. Although, at steady state, furin is predominantly found in the trans-Golgi network (TGN), it also cycles between the TGN and the cell surface. Recently, the cytoplasmic tail of furin has been shown to be sufficient for its localization to the TGN. Within the cytoplasmic domain, there are Ser residues, which we now show are sites for phosphorylation by casein kinase II in vitro, and a Tyr-containing sequence, both of which have been shown to be important for other TGN proteins to localize to this compartment. In the present study, we show by site-directed mutagenesis that these residues are important for TGN localization and recycling of furin. Mutation of the Ser residues abrogated the TGN localization. By contrast, mutation of the Tyr residue did not affect the TGN localization but impaired the internalization from the plasma membrane. These observations suggest that distinct cytoplasmic determinants are responsible for retention in the TGN and retrieval from the cell surface of furin. PMID- 7499346 TI - Human cathepsin G lacking functional glycosylation site is proteolytically processed and targeted for storage in granules after transfection to the rat basophilic/mast cell line RBL or the murine myeloid cell line 32D. AB - The neutral protease cathepsin G belongs to a family of hematopoietic serine proteases stored in the azurophil granules of the neutrophil granulocyte. To investigate the function of asparagine-linked carbohydrates in neutrophil serine proteases, we constructed a mutant cDNA, coding for human cathepsin G deficient of a functional glycosylation site, for use in a transgenic cellular model. Wild type and mutant cDNA were stably expressed in the rat basophilic/mast cell line RBL and in the murine myeloblast-like cell line 32D. Biosynthetic labeling, followed by immunoprecipitation, SDS-polyacrylamide gel electrophoresis, and fluorography, showed that carbohydrate-deficient cathepsin G was synthesized as a 29-kDa proform in both cell lines. The proform was proteolytically processed into a stable form with an apparent molecular mass of 27.5 kDa, indicating removal of the carboxyl-terminal prodomain. The mutant cathepsin G was enzymatically activated as determined by acquisition of affinity to aprotinin, a serine protease inhibitor. As for wild type cathepsin G, small amounts of the unprocessed form of the mutated enzyme were released from the cells, while the major part was transferred to a granular compartment as demonstrated by subcellular fractionation. Thus, neither processing leading to enzymatic activation nor granular sorting was obviously affected by the lack of oligosaccharides on the mutant cathepsin G. Our results therefore indicate that glycosylation is not essential for these processes. In addition to the previously utilized cell line RBL, we propose the 32D cell line as a suitable cellular model for transgenic expression of human neutrophil serine proteases. PMID- 7499347 TI - Cell cycle-dependent regulation of the cyclin B1 promoter. AB - Cyclin B1 mRNA expression varies through the cell cycle with its peak in G2/M. In cycling mammalian cells, its lowest level is in G1 with a steady increase in S until a level 50-fold greater than that in G1 is reached. In order to characterize the transcriptional component to this variation in expression, we cloned the upstream region 872 base pairs upstream from the start site of the cyclin B1 gene and have demonstrated that it confers cell cycle-dependent regulation onto two reporter genes, both chloramphenicol acetyltransferase and luciferase. Its activity was 25-fold greater in G2/M than in G1 in HeLa cells with intermediate activity in S. This cyclical activity could be seen with sequences encompassing only 90 base pairs upstream from the start site. Protein binding to this region was demonstrated using electrophoretic mobility shift assays, and the binding profiles appeared to vary depending upon the phase of the cycle in which the extracts are made. Thus, transcriptional control plays an important role in determining cyclin B1 mRNA levels, and cell cycle-dependent activity is regulated through interactions with the region 90 bases upstream from the start site. PMID- 7499345 TI - Cloning of a cDNA for a second retinol dehydrogenase type II. Expression of its mRNA relative to type I. AB - A retinol dehydrogenase, RoDH(1), which recognizes holo-cellular retinol-binding protein (CRBP) as substrate, has been cloned, expressed, and identified as a short-chain dehydrogenase/reductase (Chai, X., Boerman, M. H. E. M., Zhai, Y., and Napoli, J. L. (1995) J. Biol. Chem. 270, 3900-3904). This work reports the cloning and expression of a cDNA encoding a RoDH isozyme, RoDH(II). The predicted amino acid sequence verifies RoDH(II) as a short-chain dehydrogenase/reductase, 82% identical with RoDH(I). RoDH(II) recognized the physiological form of retinol as substrate, CRBP, with a Km of 2 mM. Similar to microsomal RoDH and RoDH(I), RoDH(II) had higher activity with NADP rather than NAD, was stimulated by ethanol and phosphatidyl choline, was not inhibited by the medium-chain alcohol dehydrogenase inhibitor 4-methylpyrazole, but was inhibited by phenylarsine oxide and the short-chain dehydrogenase/reductase inhibitor carbenoxolone. Northern blot analysis detected RoDH(I) and RoDH(II) mRNA only in rat liver, but RNase protection assays revealed RoDH(I) and RoHD(II) mRNA in kidney, lung, testis, and brain. These data indicate that short-chain dehydrogenases/reductase isozymes expressed tissue-distinctively catalyze the first step of retinoic acid biogenesis from the physiologically most abundant substrate, CRBP. PMID- 7499348 TI - Wortmannin, an inhibitor of phosphoinositide 3-kinase, inhibits transcytosis in polarized epithelial cells. AB - Wortmannin, an inhibitor of phosphoinositide 3-kinase, inhibits both basolateral to apical and apical to basolateral transcytosis of ricin in Fisher rat thyroid (FRT) cells by 50% at 100 nM in a continuous transcytosis assay. In MDCK cells, a similar effect of wortmannin on basolateral to apical transcytosis of ricin was found, whereas apical to basolateral transcytosis was inhibited to a lesser degree. Transcytosis of dimeric IgA in MDCK cells expressing the polymeric immunoglobulin receptor was also reduced to 50% of controls, suggesting that wortmannin inhibits membrane translocation rather than sorting of specific proteins in the transcytotic pathway. This effect of wortmannin is selective, however, in that endocytosis at the basolateral domain and recycling at both the basolateral and apical membrane domains are unaffected, and apical endocytosis and apical secretion are only moderately reduced. We have shown previously that cAMP stimulates a late stage in basolateral to apical transcytosis in MDCK cells through activation of protein kinase A (Hansen, S. H., and Casanova, J.E. (1994) J. Cell Biol. 126, 677-687). Elevation of cellular cAMP still induced a 100% increase in transcytosis in wortmannin-treated cells, but transcytosis was no longer increased when compared to cells which received no drugs. In contrast, in experiments using a 17 degrees C block to accumulate ricin internalized from the basolateral surface in the apical compartment of MDCK cells, wortmannin had little effect on the stimulation of transcytosis by activators of protein kinase A observed under these conditions. The data thus suggest the existence of a wortmannin-sensitive step in the transcytotic pathway, positioned after endocytosis but prior to translocation into the protein kinase A-sensitive apical compartment, implying a role for phosphoinositide 3-kinase in an intermediate step in transcytosis in polarized epithelial cells. PMID- 7499349 TI - Translational control by influenza virus. Identification of cis-acting sequences and trans-acting factors which may regulate selective viral mRNA translation. AB - We have shown that sequences contained within the viral mRNA 5'-untranslated region (UTR) played a critical role in directing selective influenza viral mRNA translation. We therefore attempted to identify transacting factors that may regulate viral mRNA translation through interactions with the 5'-UTR and at the same time map the precise sequences to which these factors bind. We can now demonstrate that multiple cellular proteins interact with influenza viral but not cellular 5'-UTRs using gel mobility shift and UV cross-linking analyses. Gel supershift studies revealed that the La autoantigen was one of the cellular proteins that interacted with the viral 5'-UTR. Utilizing mutants of the viral mRNA 5' UTR, we have determined that sequences within the very 5'-conserved region and nucleotides immediately 3' are necessary but not always sufficient for binding certain cellular proteins. Northwestern analysis showed the binding of a distinct subset of cellular proteins to the viral 5'-UTR, but also demonstrated interactions of the viral nonstructural protein NS1. Gel shift analysis with purified recombinant NS1 confirmed the binding of the viral protein to a specific region of the viral 5'-UTRs. A model describing the possible role of these cellular and viral RNA-binding proteins in regulating influenza virus mRNA translation will be discussed. PMID- 7499350 TI - Angiotensin II activates at least two tyrosine kinases in rat liver epithelial cells. Separation of the major calcium-regulated tyrosine kinase from p125FAK. AB - In rat liver epithelial cell lines (WB or GN4), angiotensin II (Ang II) stimulates cytosolic tyrosine kinase activity, in part, through a calcium dependent mechanism. In other cell types, selected hormones that activate Gi- or Gq-coupled receptors stimulate the soluble tyrosine kinase, p125FAK. Immunoprecipitation of p125FAK from Ang II-activated GN4 cells demonstrated a doubling of p125FAK kinase activity. However, an additional Ang II-activated tyrosine kinase (or kinases) representing the majority of the total activity was detected when the remaining cell lysate, immunodepleted of p125FAK, was reimmunoprecipitated with an anti-phosphotyrosine antibody. Cytochalasin D pretreatment blocks G-protein receptor-dependent tyrosine phosphorylation in Swiss 3T3 cells. While cytochalasin D decreased the Tyr(P) content of 65-75-kDa substrates in Ang II-treated GN4 cells, it did not diminish tyrosine phosphorylation of 115-130-kDa substrates, again suggesting activation of at least two tyrosine kinase pathways in GN4 cells. To search for additional Ang II activated enzymes, we used molecular techniques to identify 20 tyrosine kinase sequences in these cell lines. None was the major cytosolic enzyme activated by Ang II. Specifically, JAK2, which had been shown by others to be stimulated by Ang II in smooth muscle cells, was not activated by Ang II in GN4 cells. Finally, we purified Tyr(P)-containing tyrosine kinases from Ang II-treated cells, using anti-Tyr(P) and ATP affinity resins; 80% of the tyrosine kinase activity migrated as a single 115-120-kDa tyrosine-phosphorylated protein immunologically distinct from p125FAK. In summary, Ang II activates at least two separate tyrosine kinases in rat liver epithelial cells; p125FAK and a presumably novel, cytosolic 115-120 kDa protein referred to as the calcium-dependent tyrosine kinase. PMID- 7499351 TI - Nuclear factor binding sites in human beta globin IVS2. AB - The second intron of the human beta globin gene (beta IVS2) has been previously identified as a region required for proper expression of beta globin. To further characterize this region, we have footprinted the entire beta IVS2 and have analyzed regions of interest by electrophoretic mobility shift assay. Through these studies we have identified four utilized binding sites for the erythroid regulatory factor GATA-1, two sites bound by general transcription factor Oct-1, two sites bound by the nuclear matrix attachment DNA binding protein special A-T rich binding protein 1, and a site bound by a potential homeobox protein. Additionally, we have found several factors displaying temporal or tissue specificity by electrophoretic mobility shift assay, which may be potentially involved in the regulation of beta globin expression. These proteins are not supershifted by antibodies to factors important in erythroid regulation such as GATA-1, NFE-2, or YY1, or by antibodies against more general transcription factors. PMID- 7499352 TI - Schizosaccharomyces pombe thiamine pyrophosphokinase is encoded by gene tnr3 and is a regulator of thiamine metabolism, phosphate metabolism, mating, and growth. AB - The Schizosaccharomyces pombe gene tnr3 has been genetically defined as a negative regulator of genes involved in thiamine metabolism (Schweingruber, A. M., Frankhauser, H., Dlugonski, J., Steinmann-Loss, C., and Schweingruber, M. E. (1992) Genetics 130, 445-449). We have isolated and sequenced the gene and show that it codes for a putative protein of 569 amino acids which exhibits, in its carboxyl-terminal half, good homology to Saccharomyces cerevisiae thiamine pyrophosphokinase (TPK). tnr3 mutants have reduced levels of intracellular thiamine diphosphate, show impaired TPK activity, which is enhanced by introducing the tnr3 wild type gene on a plasmid, and can be complemented by the S. cerevisiae TPK-encoding gene TH180. These data strongly suggest that tnr3 encodes S. pombe TPK. We present evidence that TPK also acts as a negative regulator for gene pho1, which is derepressed when cells are starved for phosphate and show that in contrast to wild type cells, tnr3 mutants mate constitutively in response to thiamine, indicating that TPK is also involved in regulation of mating. Disruption of the tnr3 gene is lethal, and a tnr3 mutant expressing only residual TPK activity grows slowly and shows aberrant morphology. PMID- 7499353 TI - In vitro evidence for growth regulation of tRNA gene transcription in yeast. A role for transcription factor (TF) IIIB70 and TFIIIC. AB - We report in vitro studies showing that tRNA gene transcription in yeast is down regulated during the transition from logarithmic to stationary phase growth. Transcription in a postdiauxic (early stationary) phase extract of a wild-type strain decreased 3-fold relative to a log phase extract. This growth stage related difference in transcription was amplified to 20-fold in extracts of a strain containing a mutation (pcf1-4) in the 131-kDa subunit of TFIIIC. The reduction in transcription activity in both wild-type and mutant postdiauxic phase extracts was correlated with a decrease in the amount of TFIIIB70, the limiting factor in these extracts. However, the 3.7 +/- 0.5-fold decrease in amount of TFIIIB70 in mutant extracts does not, by itself, account for the 20 fold decrease in transcription. Accordingly, transcription in the mutant postdiauxic phase extract could be reconstituted to a level equal to the mutant log phase extract by the addition of two components, TFIIIB70 and TFIIIC. Addition of TFIIIB70 increased transcription 10-fold, while a 2-fold effect of TFIIIC was seen at saturating levels of TFIIIB70. The data suggest that both TFIIIB70 and TFIIIC play a role in coordinating the level of polymerase III transcription with cell growth rate. PMID- 7499354 TI - Glucocorticoids increase osteopontin expression in cardiac myocytes and microvascular endothelial cells. Role in regulation of inducible nitric oxide synthase. AB - In heart muscle, the cytokine-inducible isoform of nitric oxide synthase (NOS2) is expressed in both cardiac myocytes and microvascular endothelial cells (CMEC). mRNA levels for both NOS2 and for osteopontin, a multifunctional extracellular matrix phosphoprotein containing and RGD integrin binding domain, are increased in cardiac muscle following intraperitoneal injection of adult rats with lipopolysaccharide. In vitro, interleukin-1 beta and interferon-gamma increased osteopontin mRNA levels in CMEC as well as NOS2 expression in both CMEC and cardiac myocytes. However, osteopontin mRNA levels in heart muscle in vivo, and in cardiac myocytes and CMEC in vitro, also are increased 10-30-fold by the synthetic glucocorticoid dexamethasone, an agent that suppresses cytokine induction of NOS2 in both cell types. The hexapeptide GRGDSP, which interrupts binding of RGD-containing proteins to cell surface integrins, increased NOS2 mRNA, while a synthetic osteopontin peptide analogue decreased NOS2 mRNA and protein levels in both cytokine-pretreated cardiac myocytes and CMEC cultures. Also, transfection with a full-length antisense-osteopontin cDNA in cytokine pretreated CMEC decreased endogenous osteopontin mRNA and increased NOS2 mRNA levels. These results suggest that osteopontin could regulate the location and extent of NOS2 induction in the heart. Increased expression of osteopontin also may be one mechanism by which glucocorticoids suppress NOS2 activity in cardiac myocytes and microvascular endothelial cells. PMID- 7499355 TI - Molecular characterization of a putative Arabidopsis thaliana copper transporter and its yeast homologue. AB - At the molecular level, little is known about the transport of copper across plant membranes. We have isolated an Arabidopsis thaliana cDNA by complementation of a mutant (ctr1-3) of Saccharomyces cerevisiae defective in high affinity copper uptake. This cDNA codes for a highly hydrophobic protein (COPT1) of 169 amino acid residues and with three putative transmembrane domains. Most noteworthy, the first 44 residues display significant homology to the methionine- and histidine-rich copper binding domain of three bacterial copper binding proteins, among these a copper transporting ATPase. Mutant yeast cells expressing COPT1 exhibit nearly wild type behavior with regard to growth on a nonfermentable carbon source and resistance to copper and iron starvation. Expression of COPT1 is also associated with an increased sensitivity to copper toxicity. Additionally, COPT1 shows significant homology to an open reading frame of 189 amino acid residues on yeast chromosome VIII. This gene (CTR2) may encode an additional yeast metal transporter able to mediate the uptake of copper. A mutation in CTR2 displays a higher level of resistance to toxic copper concentrations. Overexpression of CTR2 provides increased resistance to copper starvation and is also associated with an increased sensitivity to copper toxicity. The amino acid sequence of CTR2, like Arabidopsis COPT1, contains three potential transmembrane domains. Taken together, the data suggest that a plant metal transporter, which is most likely involved in the transport of copper, has been identified. PMID- 7499356 TI - Mutually exclusive neuronal expression of peptides encoded by the FMRFa gene underlies a differential control of copulation in Lymnaea. AB - An innovative method, direct peptide profiling of small samples of nervous tissue by matrix-assisted laser desorption ionization mass spectrometry, in combination with peptide characterization, immunocytochemistry in conjunction with specific neuronal labeling by backfilling of the penis nerve, and bioassay of peptides was used to study the intrinsic neuronal expression patterns of distinct sets of related FMRFa peptides and their significance for the organization of male copulation behavior in the mollusk, Lymnaea stagnalis. Previous studies indicate that the sets of FMRFa-related and GDPFLRFa-related peptides are encoded by two alternatively spliced transcripts of the single FMRFa gene. Direct mass spectrometry revealed that both FMRFa-related and GDPFLRFa-related peptides are present in the penis nerve, the sole nerve that innervates the penis complex. Accordingly, authentic FMRFa, GDPFLRFa, and related peptides were purified from the penis complex. The loci of synthesis of FMRFa and related peptides could be traced to the right cerebral ventral lobe, those of GDPFLRFa and related peptides to the B group neurons in the right parietal ganglion and to a few unidentified neurons in the right pleural ganglion. Notwithstanding their related structures, the two sets of peptides have distinctly different actions on the penis retractor muscle. PMID- 7499357 TI - Protein kinase C: structure, function, and regulation. PMID- 7499358 TI - Participation of reactive oxygen species in the lysophosphatidic acid-stimulated mitogen-activated protein kinase kinase activation pathway. AB - Recent evidence suggests that reactive oxygen species (ROS) may function as second messengers in intracellular signal transduction pathways. We explored the possibility that ROS were involved in lysophosphatidic acid (LPA)-induced mitogen activated protein (MAP) kinase signaling pathway in HeLa cells. Antioxidant N acetylcysteine inhibited the LPA-stimulated MAP kinase kinase activity. Direct exposure of HeLa cells to hydrogen peroxide resulted in a concentration- and time dependent activation of MAP kinase kinase. Inhibition of catalase with aminotriazole enhanced the effect of LPA on induction of MAP kinase kinase. Further, LPA stimulated ROS production in HeLa cells. These findings suggest that ROS participate in the LPA-elicited MAP kinase signaling pathway. PMID- 7499360 TI - Hormonal regulation of the p53 tumor suppressor protein in T47D human breast carcinoma cell line. AB - Under normal culturing conditions, the T47D human breast cancer cell line expresses progesterone receptor constitutively and is responsive to estrogen. Because the tumor suppressor protein p53 plays a central role in determining genetic stability and cell proliferation, we have examined the effects of 17 beta estradiol, the synthetic progestin R5020, and the antiprogestin RU486 on the levels of this protein in T47D cells. Western blot analysis of cellular extracts, performed with a monoclonal antibody capable of quantitatively supershifting a specific p53-p53 response element complex in a gel mobility shift assay, detected a single immunoreactive band representing p53. When cells were grown for 4-5 days in culture medium containing charcoal-treated fetal calf serum, p53 levels declined to 10% of the level seen in the control (no charcoal treatment) group. Supplementation of culture medium containing charcoal-treated calf serum with 0.1 1 nM 17 beta-estradiol restored p53 to its normal levels. A 4-day treatment of cells with R5020 or RU486 lowered the p53 levels in cells grown in normal culturing conditions to 15 and 30% of control levels, respectively. R5020 and RU486 treatments also caused down-regulation and/or hyperphosphorylation of the progesterone receptor, which correlated with the down-regulation of p53. These observations by estradiol while R5020 down-regulates this protein. Since estradiol is known to promote cell proliferation, the induction of p53 observed in this study leads us to propose that estradiol stimulates p53 to regulate proliferation of T47D cells in culture. PMID- 7499359 TI - Transcriptional repression of p53 by human T-cell leukemia virus type I Tax protein. AB - The human T-cell leukemia virus type I oncoprotein Tax transcriptionally deregulates a wide variety of viral and cellular genes. Tax deregulation of gene expression is mediated through interaction with a variety of structurally unrelated cellular transcription factors, as Tax does not bind DNA in a sequence specific manner. Although most of these cellular transcription factors have been shown to mediate activation by Tax, we have recently demonstrated that members of the basic helix-loop-helix (bHLH) family of transcription factors, which play a critical role in progression through the cell cycle, mediate repression by Tax. In this report, we examined whether Tax might repress transcription of the tumor suppressor p53, as the p53 gene has recently been demonstrated to be regulated by the bHLH protein c-Myc. Furthermore, loss or inactivation of the p53 gene has been shown to be causally associated with oncogenic transformation. We show that Tax represses transcription of the p53 gene and that this repression is dependent upon the bHLH recognition element in the p53 promoter. Together, these results suggest that Tax may promote malignant transformation through repression of p53 transcription. PMID- 7499361 TI - Interaction of Phe8 of angiotensin II with Lys199 and His256 of AT1 receptor in agonist activation. AB - The acidic pharmacophores of selective ligands bind to Lys199 and His256 of the AT1 receptor (Noda, K., Saad, Y., Kinoshita, A., Boyle, T. P., Graham, R. M., Husain, A., and Karnik, S. (1995) J. Biol. Chem. 270, 2284-2289). In this report we examine how interactions between these residues and agonists activate inositol phosphate production in transiently transfected COS-1 cells. [Sar1] angiotensin (Ang II) II and [Sar1]Ang II-amide stimulated a 5-fold inositol phosphate response from wild-type AT1 receptor. The peptide antagonist [Sar1,Ile8]Ang II and the non-peptide agonist L-162,313 produced a partial but saturating response. Stimulation of wild-type receptor by [Sar1]Ang II-amide and the mutant K199Q and K199A receptors by [Sar1]Ang II demonstrates that AT1 receptor activation is not critically dependent on the ion-pairing of the alpha-COOH group of Ang II with Lys199. The mutation of His256 produced diminished inositol phosphate response without commensurate change in binding affinity of ligands. The His256 side chain is critical for maximal activation of the AT1 receptor, although isosteric Gln substitution is sufficient for preserving the affinity for Phe8-substituted analogues of [Sar1]Ang II. Therefore, AT1 receptor activation requires interaction of Phe8 side chain of Ang II with His256, which is achieved by docking the alpha-COOH group of Phe8 to Lys199. Furthermore, non-peptide agonists interact with Lys199 and His256 in a similar fashion. PMID- 7499362 TI - Identification of the site of interaction of the 14-3-3 protein with phosphorylated tryptophan hydroxylase. AB - The 14-3-3 protein family plays a role in a wide variety of cell signaling processes including monoamine synthesis, exocytosis, and cell cycle regulation, but the structural requirements for the activity of this protein family are not known. We have previously shown that the 14-3-3 protein binds with and activates phosphorylated tryptophan hydroxylase (TPH, the rate-limiting enzyme in the biosynthesis of neurotransmitter serotonin) and proposed that this activity might be mediated through the COOH-terminal acidic region of the 14-3-3 molecules. In this report we demonstrate, using a series of truncation mutants of the 14-3-3 eta isoform expressed in Escherichia coli, that the COOH-terminal region, especially restricted in amino acids 171-213, binds indeed with the phosphorylated TPH. This restricted region, which we termed 14-3-3 box I, is one of the structural regions whose sequence is highly conserved beyond species, allowing that the plant 14-3-3 isoform (GF14) could also activate rat brain TPH. The 14-3-3 box I is the first functional region whose activity has directly been defined in the 14-3-3 sequence and may represent a common structural element whereby 14-3-3 interacts with other target proteins such as Raf-1 kinase. The result is consistent with the recently published crystal structure of this protein family, which suggests the importance of the negatively charged groove like structure in the ligand binding. PMID- 7499363 TI - Signaling of ambient pH in Aspergillus involves a cysteine protease. AB - In Aspergillus nidulans, the regulation of gene expression in response to changes in ambient pH is mediated by the PacC zinc finger transcriptional regulator. At alkaline ambient pH, PacC is proteolytically processed to a functional form serving as an activator of alkaline-expressed genes and a repressor of acid expressed genes. This activation of PacC occurs in response to a signal mediated by the products of the pal genes. Thus, the products of the palA, -B, -C, -F, -H, and -I genes constitute an alkaline ambient pH signal transduction pathway. How the pal signal transduction pathway senses ambient pH and transduces a signal to trigger PacC processing is a fascinating unresolved problem. We have cloned and sequenced the palB gene. The predicted palB gene product has similarity to the catalytic domain of the calpain family of calcium-activated cysteine proteases. We have shown, however, that the PalB protein does not catalyze the final step of proteolytic processing of PacC. PMID- 7499364 TI - Molecular cloning of a novel laminin chain, alpha 5, and widespread expression in adult mouse tissues. AB - We have identified a fifth member of the alpha subfamily of vertebrate laminin chains. Sequence analysis revealed a close relationship of alpha 5 to the only known Drosophila alpha chain, suggesting that the ancestral alpha gene was more similar to alpha 5 than to alpha 1-4. Analysis of RNA expression showed that alpha 5 is widely expressed in adult tissues, with highest levels in lung, heart, and kidney. Our results suggest that alpha 5 may be a major laminin chain of adult basal laminae. PMID- 7499365 TI - Interleukins 2, 4, 7, and 15 stimulate tyrosine phosphorylation of insulin receptor substrates 1 and 2 in T cells. Potential role of JAK kinases. AB - The signaling molecules insulin receptor substrate (IRS)-1 and the newly described IRS-2 (4PS) molecule are major insulin and interleukin 4 (IL-4) dependent phosphoproteins. We report here that IL-2, IL-7, and IL-15, as well as IL-4, rapidly stimulate the tyrosine phosphorylation of IRS-1 and IRS-2 in human peripheral blood T cells, NK cells, and in lymphoid cell lines. In addition, we show that the Janus kinases, JAK1 and JAK3, associate with IRS-1 and IRS-2 in T cells. Coexpression studies demonstrate that these kinases can tyrosine phosphorylate IRS-2, suggesting a possible mechanism by which cytokine receptors may induce the tyrosine phosphorylation of IRS-1 and IRS-2. We further demonstrate that the p85 subunit of phosphoinositol 3-kinase associates with IRS 1 in response to IL-2 and IL-4 in T cells. Therefore, these data indicate that IRS-1 and IRS-2 may have important roles in T lymphocyte activation not only in response to IL-4, but also in response to IL-2, IL-7, and IL-15. PMID- 7499367 TI - Site-directed mutagenesis of the yeast PMA1 H(+)-ATPase. Structural and functional role of cysteine residues. AB - The yeast plasma-membrane H(+)-ATPase contains nine cysteines, three in presumed transmembrane segments (Cys-148, Cys-312, and Cys-867) and the rest in hydrophilic regions thought to be exposed at the cytoplasmic surface (Cys-221, Cys-376, Cys-409, Cys-472, Cys-532, and Cys-569). To gather new functional and structural information, we have studied the yeast ATPase by cysteine mutagenesis. It proved possible to replace seven of the nine cysteines by alanine, one at a time, without any significant decrease in ATP hydrolysis or ATP-dependent proton pumping. In the remaining two cases (Cys-409 and Cys-472), there were small but reproducible effects; the results clearly indicated, however, that no single Cys is required for activity and that, if a disulfide bridge is formed in the yeast ATPase, it does not play an obligatory structural or functional role. Next, multiple mutants were constructed to ask how many Cys residues could be replaced simultaneously while leaving a fully functional enzyme. After substitution of all "membrane" Cys (Cys-148, Cys-312, and Cys-867) together with two non-conserved Cys located in hydrophilic regions (Cys-221 and Cys-569), there were no significant abnormalities in expression (87%) or activity (89% ATP hydrolysis/93% H+ pumping) of the mutant protein. Replacement of two additional cysteines (Cys 376 near the phosphorylation site and Cys-532, in or near the ATP-binding site) caused a drop in expression (to 54%), although the corrected hydrolytic and H+ pumping activities were still normal. When Cys-472 was also mutated, the corrected activity fell to 44% hydrolysis/47% pumping; finally, substitution of Cys-409 to give a "cysteine-free" ATPase led to a very poorly expressed and poorly active enzyme. Brief exposure of the "one-cysteine" and "two-cysteine" ATPases to trypsin revealed a normal pattern of degradation, but there was a slight impairment in the ability of vanadate to protect against proteolysis. Thus, although single Cys replacements are tolerated well by the yeast ATPase, multiple replacements are progressively more harmful, suggesting that they cause small but additive perturbations of protein folding. PMID- 7499368 TI - Contribution of antibody heavy chain CDR1 to digoxin binding analyzed by random mutagenesis of phage-displayed Fab 26-10. AB - We constructed a bacteriophage-displayed library containing randomized mutations at H chain residues 30-35 of the anti-digoxin antibody 26-10 Fab to investigate sequence constraints necessary for high affinity binding in an antibody of known crystal structure. Phage were selected by panning against digoxin and three C-16 substituted analogues. All antigen-positive mutants selected using other analogues also bound digoxin. Among 73 antigen-positive clones, 26 different nucleotide sequences were found. The majority of Fabs had high affinity for digoxin (Ka 3.4 x 10(9) M-1) despite wide sequence diversity. Two mutants displayed affinities 2- and 4-fold higher than the parental antibody. Analysis of the statistical distribution of sequences showed that highest affinity binding occurred with a restricted set of amino acid substitutions at positions H33-35. All clones save two retained the parental Asn-H35, which contacts hapten and hydrogen bonds to other binding site residues in the parental structure. Positions H30-32 display remarkable diversity, with 10-14 different substitutions for each residue, consistent with high affinity binding. Thus complementarity can be retained and even improved despite diversity in the conformation of the N terminal portion of the H-CDR1 loop. PMID- 7499366 TI - A novel K+ channel beta-subunit (hKv beta 1.3) is produced via alternative mRNA splicing. AB - Voltage-gated K+ channels can form multimeric complexes with accessory beta subunits. We report here a novel K+ channel beta-subunit cloned from human heart, hKv beta 1.3, that has 74-83% overall identity with previously cloned beta subunits. Comparison of hKv beta 1.3 with the previously cloned hKv beta 3 and rKv beta 1 proteins indicates that the carboxyl-terminal 328 amino acids are identical, while unique variable length amino termini exist. Analysis of human beta-subunit cDNA and genomic nucleotide sequences confirm that these three beta subunits are alternatively spliced from a common beta-subunit gene. Co-expression of hKv beta 1.3 in Xenopus oocytes with the delayed rectifier hKv1.5 indicated that hKv beta 1.3 has unique functional effects. This novel beta-subunit induced a time-dependent inactivation during membrane voltage steps to positive potentials, induced a 13-mV hyperpolarizing shift in the activation curve, and slowed deactivation (tau = 13 +/- 0.5 ms versus 35 +/- 1.7 ms at -40 mV). Most notably, hKv beta 1.3 converted the Kv1.5 outwardly rectifying current voltage relationship to one showing strong inward rectification. These data suggest that Kv channel current diversity may arise from association with alternatively spliced Kv beta-subunits. A simplified nomenclature for the K+ channel beta subunit subfamilies is suggested. PMID- 7499370 TI - Studies into the effect of the tyrosine kinase inhibitor herbimycin A on NF-kappa B activation in T lymphocytes. Evidence for covalent modification of the p50 subunit. AB - The tyrosine kinase inhibitor herbimycin A was found to block NF-kappa B stimulation in response to interleukin-1 and phorbol 12-myristate 13-acetate in EL4.NOB-1 thymoma cells and phorbol 12-myristate 13-acetate in Jurkat T lymphoma cells. The effect appeared not to involve inhibition of tyrosine kinase activation as neither interleukin-1 nor phorbol 12-myristate 13-acetate induced major changes in tyrosine phosphorylation in EL4.NOB-1 or Jurkat cells, respectively. Herbimycin A did not interfere with I kappa B-alpha degradation, and in unstimulated cells, it modified NF-kappa B prior to chemical dissociation with sodium deoxycholate. Because herbimycin A is thiol-reactive, we suspected that the target was the p50 subunit of NF-kappa B, which has a key thiol at cysteine 62. Herbimycin A inhibited DNA binding when added to nuclear extracts prepared from stimulated cells, which were shown to contain high levels of p50. Incubation of herbimycin A with 2-mercaptoethanol attenuated the effect. Herbimycin A was also shown to react directly with p50, blocking its ability to bind to the NF-kappa B consensus sequence. However, a mutant form of p50 in which cysteine 62 was mutated to serine was insensitive to herbimycin A. Finally, we demonstrated that the compound inhibited the expression of interleukin-2 (an NF kappa B-regulated gene) in EL4.NOB-1 cells. These data therefore suggest that herbimycin A inhibits NF-kappa B by modifying the p50 subunit on cysteine 62 in the NF-kappa B complex, which blocks DNA binding and NF-kappa B-driven gene expression. The results urge caution in the use of herbimycin A as a specific tyrosine kinase inhibitor and suggest that the development of agents that selectively modify p50 may have potential as a means of inhibiting NF-kappa B dependent gene transcription. PMID- 7499369 TI - The chaperonin GroEL is destabilized by binding of ADP. AB - The urea-induced dissociation and subsequent conformational transitions of the nucleotide-bound form of GroEL were studied by light scattering, 4,4'-bis(1 anilino-8- naphthalenesulfonic acid) binding, and intrinsic tyrosine fluorescence. Magnesium ion alone (10 mM) stabilizes GroEL and leads to coordination of the structural transitions monitored by the different parameters. The midpoint of the light-scattering transition that monitored dissociation of the 14-mer with bound magnesium was raised to approximately 3 M, which is considerably higher than the ligand-free form of the protein, which exhibits a transition with a midpoint at approximately 2 M urea. Binding of ADP results in destabilization of the GroEL oligomeric structure, and complete dissociation of the 14-mer in the presence of 5 mM ADP occurs at about 2 M urea with the midpoint of the transition at approximately 1 M urea. The same destabilization by ADP and stabilization by Mg2+ were seen when the conformation was followed by the intrinsic fluorescence. Complexation with the nonhydrolyzable ATP analog, 5' adenylimidodiphosphate gave an apparent stability of the quaternary structure that was between that observed with Mg2+ and that with ADP. The ADP-bound form of the protein demonstrated increased hydrophobic exposure at lower urea concentrations than the uncomplexed GroEL. In addition, the GroEL-ADP complex is more accessible for proteolytic digestion by chymotrypsin than the uncomplexed protein, consistent with a more open, flexible form of the protein. The implication of the conformational changes to the mechanism of the GroEL function is discussed. PMID- 7499371 TI - Cyanide-binding site of bd-type ubiquinol oxidase from Escherichia coli. AB - We extended our investigation on the structure of the redox centers of bd-type ubiquinol oxidase from Escherichia coli using cyanide as a monitoring probe. We found that addition of cyanide to the air-oxidized O2-bound enzyme caused appearance of an infrared C-N stretching band at 2161 cm-1 and concomitant disappearance of the 647 nm absorption band of the cytochrome d (Fe2+)-O2 species. Addition of cyanide to the air-oxidized CO-bound enzyme also resulted in disappearance of the 635 nm absorption band and the 1983.4 cm-1 C-O infrared band of the cytochrome d (Fe2+)-CO species. The resulting species had a derivative shaped electron paramagnetic resonance signal at g = 3.15. Upon partial reduction with sodium dithionite, this species was converted partly to a transient heme d (Fe3+)-C = N species having an electron paramagnetic resonance signal at gz = 2.96 and a C-N infrared band at 2138 cm-1. These observations suggest that the active site of the enzyme has a heme-heme binuclear metal center distinct from that of the heme-copper terminal oxidase and that the treatment of the air oxidized enzyme with cyanide resulted in a cyanide-bridging species with "heme d(Fe3+)-C = N-heme b595(Fe3+)" structure. PMID- 7499372 TI - Thiol-disulfide exchange of ribonuclease inhibitor bound to ribonuclease A. Evidence of active inhibitor-bound ribonuclease. AB - Ribonuclease Inhibitor (RI) has been purified from pig testis. It contains 30 half-cystines whose oxidation affects its ability to bind and inhibit ribonuclease (RNase). By N-terminal sequence analyses testis RI showed to be identical to that from porcine liver, for which a characteristic all-or-none type of SH-oxidation by 5,5'-dithiobis(2-nitrobenzoic acid) (DTNB) has been reported (Fominaya, J.M., and Hofsteenge, J. (1992) J. Biol. Chem. 257, 24655-24660). Under comparable reaction conditions, testis RI bound to RNase A did not exhibit this particular type of oxidation; instead, bound RI got intermediate oxidation degrees (up to 14 thiols oxidized per RI moiety) without dissociating from RNase. Moreover, RNase bound to partially oxidized RI was able to express some (15%) of its potential activity (active complex). Only when DTNB treatments accounted for complex dissociation (> 14 thiols oxidized per RI moiety) the released RI molecules exhibited the all-or-none oxidation behavior. By both kinetic and circular dichroism analyses, conformational changes have been evidenced for the transition from the inactive to the active form of RI-RNase complex. Relaxation of RI-RNase binding without major alterations in RI structure is proposed as responsible for complex activation. The results are discussed in terms of a model for the reversible regulation of RNase activity mediated by the redox status of RI. PMID- 7499373 TI - Involvement of DNase II in nuclear degeneration during lens cell differentiation. AB - The characterization of DNase II and DNase I activity was undertaken to discriminate their different roles in physiological nuclear degradation during lens fiber cell differentiation. The activity of both nucleases determined in a new assay allows to discriminate DNase II from DNase I in the same extract. In fibers, both types of nuclease activities are found and appear higher than in epithelial cells. Specific polyclonal antibodies directed against these two nucleases reveal by Western blot analysis the presence of various DNase isoforms. DNase II like-nuclease, present in fibers, is represented by three major bands (60,23, and 18 kDa), which are not detected, at least for two of them (60 and 23 kDa), in epithelial cells. DNase I like-nuclease pattern in fiber cells shows a single 32-kDa band, while several bands can be detected in epithelial cells. Immunocytochemistry studies show both nucleases present in lens cell sections. DNase II is, as usual, in cytoplasm of epithelial cells, but it appears strikingly concentrated in the nuclei of fibers. DNase I is always concentrated in nuclei of epithelial and fiber cells. DNA degradation observed in agarose gels shows that DNase II-activating medium cleaves the DNA from fiber cells more efficiently than DNase I-activating buffer. In addition, DNase II antibody is able to prevent this degradation. These results suggest a specific involvement of DNase II in nuclear degradation during lens cell differentiation. PMID- 7499374 TI - Chemical modification of the N-10 ribityl side chain of flavins. Effects on properties of flavoprotein disulfide oxidoreductases. AB - Three flavin derivatives modified at the 2'-position of the flavin N-10 ribityl side chain were synthesized: arabinoflavin, 2'-F-2'-deoxyarabinoflavin, and 2' deoxyriboflavin. These were converted to the FAD level with FAD synthetase. Apoproteins of lipoamide dehydrogenase, glutathione reductase, and mercuric reductase, a family of flavoprotein oxidoreductases, were reconstituted with these flavins. Significant reduction of the catalytic activities was observed with the modified enzymes. During anaerobic reduction of the modified enzymes with substrate or dithiothreitol, decreased thermodynamic stability of the two electron reduced enzyme forms (EH2) and the accumulation of the four-electron reduced forms (EH4) noted. This effect was more pronounced in case of arabino-FAD reconstituted enzymes than with the other two. It was found that NAD+ binding influences the interaction between the flavin and the reduced disulfide in the 2' F-arabino-FAD-lipoamide dehydrogenase, presumably by altering the relative oxidation-reduction potentials. 19F NMR data were obtained for different forms of the 2'-F-arabino-FAD-lipoamide dehydrogenase, which suggest marked conformational changes from one form to the other. The 19F NMR data for the oxidized forms of all three 2'-F-arabino-FAD proteins suggest that the fluorine experiences very similar chemical environments at the active sites. PMID- 7499375 TI - Shaker K+ channel T1 domain self-tetramerizes to a stable structure. AB - The potassium channel T1 domain plays an important role in the regulated assembly of subunit proteins. We have examined the assembly properties of the Shaker channel T1 domain to determine if the domain can self-assemble, the number of subunits in a multimer, Ns and the mechanism of assembly. High pressure liquid chromatography (HPLC) size exclusion chromotography (SEC) separates T1 domain proteins into two peaks. By co-assembly assays, these peaks are identified to be a high molecular weight assembled form and a low molecular weight monomeric form. To determine the Ns of the assembled protein peak on HPLC SEC, we first cross linked the T1 domain proteins and then separated them on HPLC. Four evenly spaced bands co-migrate with the assembled protein peak; thus, the T1 domain assembles to form a tetramer. The absence of separate dimeric and trimeric peaks of assembled T1 domain protein suggests that the tetramer is the stable assembled state, most probably a closed ring structure. PMID- 7499376 TI - Isolation and characterization of a cDNA clone encoding a member of the Com44/Cim44 envelope components of the chloroplast protein import apparatus. AB - Many of the proteins in the chloroplast envelope play an important role in facilitating the biochemical and transport processes of the compartment. For the transport of proteins into the chloroplast, we have recently identified at least three different envelope proteins (Com44/Cim44, Com70, and Cim97) in close physical proximity to a partially translocated chimeric precursor protein (Wu, C., Seibert, F. S., and Ko, K. (1994) J. Biol. Chem. 269, 32264-32271). In this study we report the characterization of a cDNA clone encoding a member of the Com44/Cim44 envelope proteins. The combined data from nucleotide sequencing, and RNA and protein blot analyses indicate the existence of multiple forms of the 44 kDa envelope protein. Depending on the plant species examined, immunologically related protein bands with molecular masses of 42 to 46 kDa were observed. Organelle subfractionation, protease treatment, and immunomicroscopic studies together provide an indication that the immunologically-related proteins may be present in both the outer and inner envelope membranes. Co-migration of the product synthesized from the cDNA insert with a 44-kDa immunoreactive band of the chloroplast envelope, and the in vitro import results, together suggest that the in vitro synthesized 44-kDa protein is targeted to the envelope membrane without any further processing. PMID- 7499377 TI - Interaction of deoxyinosine 3'-endonuclease from Escherichia coli with DNA containing deoxyinosine. AB - By using a band mobility shift assay, deoxyinosine 3'-endonuclease, an Escherichia coli enzyme which recognizes deoxyinosine, AP site, urea residue, and base mismatches in DNA, was shown to bind tightly to deoxyinosine-containing oligonucleotide duplexes. Two distinct protein-DNA complexes were observed, the faster migrating complex (complex I, Kd = 4 x 10(-9) M) contained one molecule of deoxyinosine 3'-endonuclease, while the slower migrating complex (complex II, Kd = 4 x 10(-7) M) contained two molecules of the protein bound to every molecule of duplex DNA. The endonucleolytic activity of deoxyinosine 3'-endonuclease paralleled the formation of the complex I. Interestingly, deoxyinosine 3' endonuclease exhibited similar affinities for both the substrate and the nicked duplex product and thus remained bound to the DNA after the cleavage reaction. The formation of a stable complex required the presence of a duplex structure 5' to the deoxyinosine residue. DNase I footprinting revealed that deoxyinosine 3' endonuclease protected 4-5 nucleotides 5' to the deoxyinosine, and when complex II was formed, at least 13 nucleotides 3' to deoxyinosine were protected. Based on these results, a model is proposed for the interaction of deoxyinosine 3' endonuclease with DNA containing deoxyinosine. PMID- 7499378 TI - A proteasome from the methanogenic archaeon Methanosarcina thermophila. AB - A 645-kDa proteasome was purified from Methanosarcina thermophila which had chymotrypsin-like and peptidylglutamyl-peptide hydrolase activities and contained alpha (24-kDa) and beta (22-kDa) subunits. Processing of both subunits was suggested by comparison of N-terminal sequences with the sequences deduced from the alpha- and beta-encoding genes (psmA and psmB). Alignment of deduced sequences for the alpha and beta subunits revealed high similarity; however, the N-terminal sequence of the alpha subunit contained an additional 24 amino acids that were not present in the beta subunit. The alpha and beta subunits had high sequence identity with alpha- and beta-type subunits of proteasomes from eucaryotic organisms and the distantly related archaeon Thermoplasma acidophilum. The psmB gene was transcribed in vivo as a monocistronic message from a consensus archaeal promoter. The results suggest that proteasomes are more widespread in the Archaea than previously proposed. Southern blotting experiments suggested the presence of ubiquitin-like sequences in M. thermophila. PMID- 7499379 TI - Functional analysis of the transforming growth factor beta responsive elements in the WAF1/Cip1/p21 promoter. AB - The transforming growth factor beta s (TGF-beta s) are a group of multifunctional growth factors that inhibit cell cycle progression in many cell types. The TGF beta-induced cell cycle arrest has been partially attributed to the regulatory effects of TGF-beta on both the levels and activities of the G1 cyclins and their cyclin-dependent kinase partners. The ability of TGF-beta to inhibit the activity of these kinase complexes derives in part from its regulatory effects on the cyclin-dependent kinase inhibitors, p21/WAF1/Cip1, p27Kip1, and p15. Upon treatment of cells with TGF-beta, these three inhibitors bind to and block the activities of specific cyclin-cyclin-dependent kinase complexes to cause cell cycle arrest. Little is known, however, on the mechanism through which TGF-beta activates these cyclin-dependent kinase inhibitors. In the case of p21, TGF-beta treatment leads to an increase in p21 mRNA. This increase in p21 mRNA is partly due to transcriptional activation of the p21 promoter by TGF-beta. To further define the signaling pathways through which TGF-beta induces p21, we have performed a detailed functional analysis on the p21 promoter. Through both deletion and mutation analysis of the p21 promoter, we have defined a 10-base pair sequence that is required for the activation of the p21 promoter by TGF beta. In addition, this sequence is sufficient to drive TGF-beta-mediated transcription from a previously nonresponsive promoter. Preliminary gel shift assays demonstrate that this TGF-beta responsive element binds specifically to several proteins in vitro. Two of these proteins are the transcription factors Sp 1 and Sp-3. These studies represent the initial steps toward defining the signaling pathways involved in TGF-beta-mediated transcriptional activation of p21. PMID- 7499380 TI - Triabin, a highly potent exosite inhibitor of thrombin. AB - Triabin, a new thrombin inhibitor, has been purified from the saliva of Triatoma pallidipennis, a blood-sucking triatomine bug. It forms a noncovalent complex with thrombin at a molar ratio of 1:1, inhibits thrombin-induced platelet aggregation, and prolongs thrombin clotting time and activated partial thromboplastin time. However, it only minimally suppresses the amidolytic activity of thrombin, as measured by a chromogenic peptide substrate assay. It completely blocks trypsin-catalyzed cleavage of thrombin, probably via protection of the anion-binding exosite and inhibits the effect of thrombomodulin on thrombin in a dose-dependent fashion. These results indicate that the inhibitor is directed toward the anion-binding exosite of thrombin. The protein was partially sequenced and the information used to isolate cDNA clones from a T. pallidipennis salivary gland library. Four slightly polymorphic variants coding for mature proteins of 142 amino acids preceded by a putative leader sequence were obtained. The recombinant protein expressed in the periplasmic space of Escherichia coli has a biological activity similar to that of salivary triabin, as tested in a thrombin-induced platelet aggregation assay. In addition, recombinant triabin inhibits thrombin-catalyzed hydrolysis of fibrinogen with a Ki of about 3 pM. PMID- 7499381 TI - Thioredoxin-linked "thiol peroxidase" from periplasmic space of Escherichia coli. AB - Three different molecular masses (24, 22, and 20 kDa) of antioxidant proteins were purified in Escherichia coli. These proteins exhibited the preventive effects against the inactivation of glutamine synthetase activity and the cleavage of DNA by a metal-catalyzed oxidation system capable of generating reactive oxygen species. Their antioxidant activities were supported by a thiol reducing equivalent such as dithiothreitol. Analysis of the amino-terminal amino acid sequences and the immunoblots between 24- and 22-kDa proteins indicates that the 24-kDa protein is an intact form of the 22-kDa protein that was previously identified 22-kDa subunit (AhpC) of E. coli alkyl hydroperoxide reductase (AhpC/AhpF). We isolated and sequenced an E. coli genomic DNA fragment that encodes 20-kDa protein. Comparison of the deduced amino acid sequence of the 20 kDa protein with that of AhpC revealed no sequence homology. A search of a data bank showed that the 20-kDa protein is a new type of antioxidant enzyme. The synthesis of this novel 20-kDa protein was increased in response to oxygen stress during growth. The 20-kDa protein resides mainly in the periplasmic space of E. coli, whereas the 24-kDa AhpC resides mainly in the matrix. The 20-kDa protein was functionally linked to the thioredoxin as an in vivo thiol-regenerating system and exerted a peroxidase activity. This 20-kDa protein is thus named "thiol peroxidase," which could act as an antioxidant enzyme removing peroxides or H2O2 within the catalase- and peroxidase-deficient periplasmic space of E. coli. PMID- 7499382 TI - Interface peptides as structure-based human immunodeficiency virus reverse transcriptase inhibitors. AB - Reverse transcriptases from both human immunodeficiency viruses type 1 and 2 are obligatory dimers. A tryptophan-rich repeat motif that is highly conserved between these proteins, as well as in the reverse transcriptase from simian immunodeficiency virus, has been postulated to be involved in hydrophobic subunit interactions. A synthetic 19-mer peptide covering part of this tryptophan repeat motif was recently shown to inhibit human immunodeficiency viruses type 1 reverse transcriptase subunit dimerization (Divita, G., Restle, T., Goody, R. S., Chermann, J.-C., and Baillon, J. G. (1994) J. Biol. Chem. 269, 13080-13083). In the present study, we show that the same peptide can also inhibit human immunodeficiency virus type 2 reverse transcriptase subunit dimerization, suggesting that the same inhibitors might be used as agents against both viruses as well as against variants of human immunodeficiency virus type 1 that differ from the variant against which they were developed. Under appropriate experimental conditions, e.g. at acidic pH, this peptide is also able to induce the dissociation of the enzyme from human immunodeficiency virus type 1. PMID- 7499384 TI - Possible role for serine/threonine phosphorylation in the regulation of the heteroprotein complex between the hsp90 stress protein and the pp60v-src tyrosine kinase. AB - The abundant, cytoplasmic 90-kDa heat-shock protein associates transiently with the Rous sarcoma virus oncogenic protein tyrosine kinase, pp60v-src, directs its cellular trafficking and negatively regulates its kinase activity. Here we report that the serine/threonine phosphatase inhibitor, okadaic acid, destabilized the heat-shock protein 90-pp60v-src chaperone complex in v-src-transfected cells. Concomitant with complex destabilization by okadaic acid, phosphoserine was doubled and phosphothreonine was increased 20-fold in the heat-shock protein 90. Although phosphorylation of the total pool of immunoprecipitable pp60v-src was unchanged, okadaic acid slightly increased phosphoserine and phosphothreonine levels specifically in pp60v-src bound to heat-shock protein 90. The low level of tyrosine phosphorylation in the pp60v-src complexed with heat-shock protein 90 was further decreased by okadaic acid. Interestingly, okadaic acid-stabilized hyperphosphorylation of the heat-shock protein 90-pp60v-src complex lowered the level of pp60v-src in cell membranes, the functional location for pp60v-src. We suggest that serine/threonine phosphorylation of heat-shock protein 90 and/or pp60v-src functions as a regulatory molecular trigger to release pp60v-src from the chaperone complex at the inner surface of cell membranes. PMID- 7499383 TI - Arachidonoyl-diacylglycerol kinase. Specific in vitro inhibition by polyphosphoinositides suggests a mechanism for regulation of phosphatidylinositol biosynthesis. AB - We previously described the purification of a membrane-bound diacylglycerol kinase highly selective for sn-1-acyl-2-arachidonoyl diacylglycerols (Walsh, J. P., Suen, R., Lemaitre, R. N., and Glomset, J. A. (1994) J. Biol. Chem. 269, 21155-21164). This enzyme appears to be responsible for the rapid clearance of the arachidonate-rich pool of diacylglycerols generated during stimulus-induced phosphoinositide turnover. We have now shown phosphatidylinositol 4,5 bisphosphate to be a potent and specific inhibitor of arachidonoyl-diacylglycerol kinase. Kinetic analyses indicated a Ki for phosphatidylinositol 4,5-bisphosphate of 0.04 mol %. Phosphatidic acid also was an inhibitor with a Ki of 0.7 mol %. Other phospholipids had only small effects at these concentrations. A series of multiply phosphorylated lipid analogs also inhibited the enzyme, indicating that the head group phosphomonoesters are the primary determinants of the polyphosphoinositide effect. However, these compounds were not as potent as phosphatidylinositol 4,5-bisphosphate, indicating some specificity for the polyphosphoinositide additional to its total charge. Five other diacylglycerol kinases were activated to varying degrees by phosphatidylinositol 4,5 bisphosphate and phosphatidic acid, suggesting that inhibition by acidic lipids may be specific for the arachidonoyl-DAG kinase isoform. Given the presumed role of arachidonoyl-diacylglycerol kinase in the phosphoinositide cycle, this inhibition may represent a mechanism for polyphosphoinositides to regulate their own synthesis. PMID- 7499385 TI - Molecular properties of neuronal G-protein-activated inwardly rectifying K+ channels. AB - Four cDNA-encoding G-activated inwardly rectifying K+ channels have been cloned recently (Kubo, Y., Reuveny, E., Slesinger, P. A., Jan, Y. N., and Jan, L. Y. (1993) Nature 364, 802-806; Lesage, F., Duprat, F., Fink, M., Guillemare, E., Coppola, T., Lazdunski, M., and Hugnot, J. P. (1994) FEBS Lett. 353, 37-42; Krapivinsky, G., Gordon, E. A., Wickman, K., Velimirovic, B., Krapivinsky, L., and Clapham, D. E. (1995) Nature 374, 135-141). We report the cloning of a mouse GIRK2 splice variant, noted mGIRK2A. Both channel proteins are functionally expressed in Xenopus oocytes upon injection of their cRNA, alone or in combination with the GIRK1 cRNA. Three GIRK channels, mGIRK1-3, are shown to be present in the brain. Colocalization in the same neurons of mGIRK1 and mGIRK2 supports the hypothesis that native channels are made by an heteromeric subunit assembly. GIRK3 channels have not been expressed successfully, even in the presence of the other types of subunits. However, GIRK3 chimeras with the amino- and carboxyl-terminal of GIRK2 are functionally expressed in the presence of GIRK1. The expressed mGIRK2 and mGIRK1, -2 currents are blocked by Ba2+ and Cs+ ions. They are not regulated by protein kinase A and protein kinase C. Channel activity runs down in inside-out excised patches, and ATP is required to prevent this rundown. Since the nonhydrolyzable ATP analog AMP-PCP is also active and since addition of kinases A and C as well as alkaline phosphatase does not modify the ATP effect, it is concluded that ATP hydrolysis is not required. An ATP binding process appears to be essential for maintaining a functional state of the neuronal inward rectifier K+ channel. A Na+ binding site on the cytoplasmic face of the membrane acts in synergy with the ATP binding site to stabilize channel activity. PMID- 7499386 TI - The involvement of threonine 160 of cytochrome b of Rhodobacter sphaeroides cytochrome bc1 complex in quinone binding and interaction with subunit IV. AB - The cytochrome b subunit (subunit I) of the ubiquinolcytochrome c reductase (bc1 complex) is thought to participate in the formation of two quinone/quinol reaction centers, an oxidizing center (Qo) and a reducing center, in accordance with the quinone cycle mechanism. Threonine 160 is a highly conserved residue in a segment of subunit I that was shown to bind quinone and is placed near the putative Qo site in current models of the bc1 complex. Rhodobacter sphaeroides cells expressing bc1 complexes with Ser or Tyr substituted for Thr160 grow photosynthetically at a reduced rate, and cells expressing the mutated complexes produce an "elevated" level of the bc1 complex. The Ser substitution also affects the interaction of subunit IV with subunit I. Replacement of Thr160 by Ser results in about a 70% loss of the activity in the purified complex, whereas substitution by Tyr lowers the activity by more than 80%. Both replacements lower the apparent Km for ubiquinol. Electron paramagnetic resonance (EPR) spectroscopy shows that in the Ser substituted complex, the environments of the Rieske iron sulfur cluster in subunit III and the high potential cytochrome b (b562) in subunit I have been modified. The spectra of the Ser160 and Tyr160 iron-sulfur clusters have become redox-insensitive, with a line shape resembling that of the native complex in the fully reduced state. The EPR signal of b562 in the Ser160 complex is shifted from g = 3.50 to g = 3.52, but otherwise the line shape is very similar to the spectrum of the native complex. Most of these results are consistent with current ideas regarding the structure and function of Qo in the bc1 complex, except for the alteration of the b562 EPR feature, because this heme is not thought to be located in proximity to Qo. Immunoblotting analysis showed that the Ser or Tyr substituted complex contained significantly less than a stoichiometric amount of subunit IV. The enzymatic activity of mutated bc1 complex was found to be activable by the addition of purified subunit IV. These results indicate that Thr160 plays an important role in the structure and/or function of the bc1 complex. PMID- 7499387 TI - Molecular characterization of hasC from an operon required for hyaluronic acid synthesis in group A streptococci. Demonstration of UDP-glucose pyrophosphorylase activity. AB - Hyaluronic acid is a high molecular weight glycosaminoglycan composed of repeating subunits of glucuronic acid and N-acetylglucosamine. It is synthesized by the group A streptococcal membrane-associated enzyme hyaluronate synthase. In previous reports, the locus required for expression of hyaluronic acid, the has operon, was identified and found to consist of two genes, hasA and hasB encoding hyaluronate synthase and UDP-glucose dehydrogenase, respectively. Since a transcription terminator was not found at the end of hasB, it was the aim of this study to identify the remaining gene(s) in the has operon. By utilizing the Tn1000 method of DNA sequencing and inverse polymerase chain reaction, hasC, the third gene in the has operon was shown to be 915 base pairs in length (304 amino acids) and located 192 base pairs downstream of hasB. Sequence similarities to other genes suggested that hasC encodes UDP-glucose pyrophosphorylase. Overexpression of hasC using isopropyl-1-thio-beta-D-galactopyranoside induction of the T7 promoter in the pET translation system allowed for the production of bacterial extracts from Escherichia coli that possessed increased UDP-glucose pyrophosphorylase activity as compared to nondetectable levels in extracts with vector alone. In addition, expression of HasC resulted in a protein of approximately 36 kDa as shown by SDS-polyacrylamide gel electrophoresis. These data as well as complementation analysis of hasC in an E. coli galU mutant confirmed that hasC encodes UDP-glucose pyrophosphorylase. Finally, since sequence analysis identified a potential rho-independent transcription terminator at the 3-prime terminus of the gene, hasC is the third and probably the final gene in the has operon. PMID- 7499388 TI - Expression and functional analysis of a novel isoform of gicerin, an immunoglobulin superfamily cell adhesion molecule. AB - We have cloned a novel cDNA of gicerin, a cell adhesion molecule belonging to the immunoglobulin superfamily. Both gicerin isoforms share the same extracellular domain, which has five immunoglobulin-like loop structures and a transmembrane domain as s-gicerin, but differ in the cytoplasmic tail domain. As the newly identified form has a larger cytoplasmic domain than the previously reported form, we refer to them as l-gicerin and s-gicerin, respectively. l-gicerin is transcribed from a distinct mRNA containing an inserted sequence not found in s gicerin mRNA which caused a frameshift for the coding region for a cytoplasmic domain. Previous studies demonstrated that gicerin showed a doublet band of 82 and 90 kDa in chicken gizzard smooth muscle. We report that the 82-kDa protein corresponds to s-gicerin and the 90-kDa protein to l-gicerin. We also found that the two gicerin isoforms are expressed differentially in the developing nervous system. Functional analysis of these gicerin isoforms in stable transfectants revealed that they had differ in their homophilic adhesion properties, as well as in heterophilic cell adhesion assayed with neurite outgrowth factor. In addition, these isoforms have neurite-promoting activity by their homophilic adhesion, but differ in their ability to promote neurite outgrowth. PMID- 7499389 TI - Defective glycolysis and calcium signaling underlie impaired insulin secretion in a transgenic mouse. AB - Pancreatic beta cells from mice that overexpress the Ca(2+)-binding protein calmodulin have a unique secretory defect that leads to chronic hyperglycemia. To further understand the molecular basis underlying this defect, we have studied signaling pathways in these beta cells. Measurements of cytosolic free Ca2+ concentration ([Ca2+]i) using fura-2 or indo-1 revealed a markedly reduced response when glucose was the stimulant. However, eliciting membrane depolarization with 50 mM K+ or the addition of the ATP-sensitive K+ (K+ ATP) channel antagonist tolbutamide restored [Ca2+]i transients to near normal levels. Electrophysiological analysis of the beta cell ion channels revealed that Ca2+ currents, delayed rectifier K+ currents, and K+ATP channel currents were similar in transgenic and nontransgenic cells, suggesting that these ion channels were able to function normally. However, whereas K+ATP channel currents in control cells were reduced by 50% by the presence of high glucose, those in transgenic cells were unaltered. Addition of tolbutamide inhibited this channel and enhanced the secretion of insulin in response to glucose for both control and transgenic cells. As these observations implicated a metabolic defect, glucose utilization, which is an indicator of glucose metabolism and ATP production in beta cells, was measured and found to be reduced by 40% in the transgenic cells. These data support the contention that excessive levels of calmodulin may compromise the ability of the beta cell to metabolize glucose and to modulate the state of the K+ATP channel, resulting in an inadequate control of the membrane potential, which collectively impair [Ca2+]i and thus insulin secretion in response to glucose. PMID- 7499391 TI - Photoaffinity labeling of a cell surface polyamine binding protein. AB - Intracellular polyamine pools are partially maintained by an active transport apparatus that is specific for and regulated by polyamines. Although mammalian transport activity has been characterized by kinetic studies, the actual protein itself has yet to be identified, purified, or cloned. As one approach to this problem, we attempted photoaffinity labeling of plasma membrane proteins using two specifically designed and synthesized polyamine conjugates as photoprobes. The first is a spermidine conjugate bearing the photoreactive moiety 4 azidosalicylic acid at the N4 position via an alkyl linkage, and the second is a norspermine conjugate with 4-azidosalicylic acid at the N4 position via an acyl linkage. Labeling of murine L1210 lymphocytic leukemia cells was carried out at 4 degrees C to promote selective alkylation of cell surface proteins. Separation of plasma membrane proteins from cells cross-linked with the N4-spermidine conjugate by SDS-polyacrylamide gel electrophoresis revealed two heavily labeled proteins at approximately 118 and approximately 50 kDa (designated p118 and p50, respectively). Band p118 was more well defined and much more intensely labeled. Analogous proteins were also observed in human U937 lymphoma cells. Specificity of labeling was strongly suggested by competition with polyamines and analogs during labeling and further indicated by the nearly identical labeling of the same protein by the N1-norspermine photoprobe but not by the unconjugated photoreagent. Neuraminidase pretreatment of L1210 cells increased mobility of the p118, suggesting that it was glycosylated and, thus, of plasma membrane origin. In transport-deficient L1210 cells, p118 and p50 were found to have a slightly higher molecular mass and were accompanied by a less distinct protein band (approximately 100 kDa). These findings indicate the presence of a polyamine binding protein at the surface of murine and human leukemia cells, which could be directly or indirectly related to the polyamine transport apparatus. PMID- 7499392 TI - Short external loops as potential substrate binding site of gamma-aminobutyric acid transporters. AB - While the gamma-aminobutyric acid (GABA) transporter GAT1 exclusively transports GABA, GAT2, -3, and -4 also transport beta-alanine. Cross-mutations in the external loops IV, V, and VI among the various GABA transporters were performed by site-directed mutagenesis. The affinity of GABA transport as well as inhibitor sensitivity of the modified transporters was analyzed. Kinetic analysis revealed that a cross-mutation in which loop IV of GAT1 was modified to resemble GAT4 resulted in increased affinity to GABA from Km = 8.7 to 2.0 microM without changing the Vmax. A cross-mutation in loop VI, which swapped the amino acid sequence of GAT2 for GAT1, decreased the affinity to GABA (Km, 35 microM). These results suggest that loops IV and VI contribute to the binding affinity of GABA transporters. A substitution of three amino acids in loop V of GAT1 by the corresponding sequence of GAT3 resulted in beta-alanine sensitivity of its GABA uptake activity. These three amino acids in loop V seem to participate in the beta-alanine binding domain of GAT3. It is suggested that those three external loops (IV, V, and VI) form a pocket in which the substrate binds to the GABA transporters. PMID- 7499393 TI - Upstream stimulatory factors bind to insulin response sequence of the fatty acid synthase promoter. USF1 is regulated. AB - Fatty acid synthase (FAS) plays a central role in de novo lipogenesis in mammals. The concentration or activity of FAS in liver and adipose tissue changes dramatically when animals are subjected to nutritional and hormonal manipulations. We previously reported that due to changes in transcription, FAS synthesis declines and increases in an insulin-dependent manner during fasting and refeeding, respectively, and that insulin administration of streptozotocin diabetic mice stimulates FAS transcription. We previously mapped the FAS insulin response sequence (IRS) to the proximal promoter region from position -71 to position -50, which contains an E-box DNA binding motif. Here, using competition gel shift assays and specific upstream stimulatory factor (USF) antibodies, we identified USF1 and USF2 as major components of complexes that bind to the FAS IRS. UV-cross-linking experiments further supported that USFs bind the FAS IRS. We also found that the amount of the 43-kDa USF1 was dramatically increased in liver of refed rats. In contrast, the amount of USF2 remained the same in liver of fasted or refed rats. Moreover, a 17-kDa protein in both fasted and refed rat liver was recognized by anti-USF1 antibodies, and this 17-kDa USF1-related protein was expressed in a manner opposite to that of the 43-kDa USF1, i.e. high in liver of fasted rats and decreased in liver of refed rats. These data suggest that the regulation of USF expression may play an important role in the regulation of FAS transcription. PMID- 7499390 TI - Peptide probe of ryanodine receptor function. Imperatoxin A, a peptide from the venom of the scorpion Pandinus imperator, selectively activates skeletal-type ryanodine receptor isoforms. AB - We have used [3H]ryanodine binding experiments and single channel recordings to provide convergent descriptions of the effect of imperatoxin A (IpTxa), a approximately 5-kDa peptide from the venom of the scorpion Pandinus imperator (Valdivia, H. H., Kirby, M. S., Lederer, W. J., and Coronado, R. (1992) Proc. Ntl. Acad. Sc. U.S.A. 89, 12185-12189) on Ca2+ release channels/ryanodine receptors (RyR) of sarcoplasmic reticulum (SR). At nanomolar concentrations, IpTxa increased the binding of [3H]ryanodine to skeletal SR and, to a lesser extent, to cerebellum microsomes. The activating effect of IpTxa on skeletal SR was Ca(2+)-dependent, synergized by caffeine, and independent of other modulators of RyRs. However, IpTxa had negligible effects on tissues where the expression of skeletal-type RyR isoforms (RyR1) is small or altogether absent, i.e. cardiac, cerebrum, and liver microsomes. Thus, IpTxa may be used as a ligand capable of discriminating between RyR isoforms with nanomolar affinity. IpTxa increased the open probability (Po) of rabbit skeletal muscle RyRs by increasing the frequency of open events and decreasing the duration of the closed lifetimes. This activating effect was dose-dependent (ED50 = 10 nM), had a fast onset, and was fully reversible. Purified RyR from solubilized skeletal SR displayed high affinity for [3H]ryanodine with a KD of 6.1 nM and Bmax of approximately 30 pmol/mg of protein. IpTxa increased [3H]ryanodine binding noncompetitively by increasing Bmax to approximately 60 pmol/mg of protein. These results suggested the presence of an IpTxa-binding site on the RyR or a closely associated regulatory protein. This site appears to be distinct from the caffeine- and adenine nucleotide-regulatory sites. IpTxa may prove a useful tool to identify regulatory domains critical for channel gating and to dissect the contribution of skeletal-type RyRs to intracellular Ca2+ waveforms generated by stimulation of different RyR isoforms. PMID- 7499394 TI - Binding sites for abundant nuclear factors modulate RNA polymerase I-dependent enhancer function in Saccharomyces cerevisiae. AB - The 190-base pair (bp) rDNA enhancer within the intergenic spacer sequences of Saccharomyces cerevisiae rRNA cistrons activates synthesis of the 35S-rRNA precursor about 20-fold in vivo (Mestel,, R., Yip, M., Holland, J. P., Wang, E., Kang, J., and Holland, M. J. (1989) Mol. Cell. Biol. 9, 1243-1254). We now report identification and analysis of transcriptional activities mediated by three cis acting sites within a 90-bp portion of the rDNA enhancer designated the modulator region. In vivo, these sequences mediated termination of transcription by RNA polymerase I and potentiated the activity of the rDNA enhancer element. Two trans acting factors, REB1 and REB2, bind independently to sites within the modulator region (Morrow, B. E., Johnson, S. P., and Warner, J. R. (1989) J. Biol. Chem. 264, 9061-9068). We show that REB2 is identical to the ABF1 protien. Site directed mutagenesis of REB1 and ABF1 binding sites demonstrated uncoupling of RNA polymerase I-dependent termination from transcriptional activation in vivo. We conclude that REB1 and ABF1 are required for RNA polymerase I-dependent termination and enhancer function, respectively, Since REB1 and ABF1 proteins also regulate expression of class II genes and other nuclear functions, our results suggest further similarities between RNA polymerase I and II regulatory mechanisms. Two rDNA enhancers flanking a rDNA minigene stimulated RNA polymerase I transcription in a "multiplicative" fashion. Deletion mapping analysis showed that similar cis-acting sequences were required for enhancer function when positioned upstream or downstream from a rDNA minigene. PMID- 7499395 TI - Activin increases phosphorylation and decreases stability of the transcription factor Pit-1 in MtTW15 somatotrope cells. AB - Activin is a polypeptide growth factor which exerts endocrine, paracrine, and autocrine effects in a variety of tissues. In the pituitary somatotrope, activin represses proliferation and growth hormone (GH) biosynthesis and secretion. We previously demonstrated that decreases in GH biosynthesis in MtTW15 somatotrope cells are due at least in part to decreased binding of the tissue-specific transcription factor, Pit-1, to the GH promoter, resulting in decreased transcription of the GH gene. The objective of the current study was to determine the extent to which activin-mediated decreases in GH transcription were the result of decreased Pit-1 activity and/or decreased Pit-1 protein content in MtTW15 cells. Activin caused rapid increases in Pit-1 phosphorylation, which were temporally correlated with decreases in GH DNA binding. Pit-1 phosphorylation preceded marked decreases in steady-state levels of Pit-1 protein. The rate of Pit-1 synthesis was only moderately decreased by activin, with a time-course similar to that observed for decreases in GH biosynthesis. However, Pit-1 stability was markedly decreased after more than 4 h of activin treatment. These data demonstrate that activin decreases GH expression in MtTW15 cells through multilevel regulation of Pit-1, which may represent a more general mechanism whereby activin and other transforming growth factor beta family members modulate gene expression through regulation of transcription factor activity as well as content. PMID- 7499396 TI - Alpha 4 beta 1 integrin-dependent cell adhesion is regulated by a low affinity receptor pool that is conformationally responsive to ligand. AB - alpha 4 beta 1 integrin (VLA-4) appears to be unique among the leukocyte integrins in that it can initiate the adhesion of circulating lymphocytes without cellular activation. It is not known how lymphocytes or other cell types maintain constitutive levels of alpha 4 beta 1 integrin activity. The current report describes a monoclonal antibody, 15/7, that recognizes a high affinity or ligand occupied conformation of beta 1 integrin. Studies with 15/7 revealed that alpha 4 beta 1 integrin-dependent adhesion of leukocytic cell lines is mediated by a population of low affinity receptors that is conformationally responsive to ligand; the 15/7 epitope could be induced by nanomolar concentrations of soluble VCAM-1 or by micromolar concentrations of a peptide derived from the type III connecting segment domain of fibronectin (as ligands for alpha 4 beta 1 integrin). The same receptors were also responsive to adhesion activating reagents, such as Mn2+, activating anti-beta 1 integrin antibodies, and phorbol myristate acetate, which induced the 15/7 epitope directly and/or decreased the concentration of ligand required for epitope induction. In addition to the responsive receptor pool, cells expressed a second population of alpha 4 beta 1 integrin that was conformationally restrained, failing to respond to ligand or to any of the activating reagents. The relative size of the responsive and inactive receptor pools, as well as the affinity of the responsive receptors, represented a stable phenotype of different cell types and played important roles in defining the cells' adhesive capacity and ligand specificity. Similar receptor populations were measured on lymphocyte subsets in whole blood. These studies provide insight into how cells maintain different constitutive levels of alpha 4 beta 1 integrin activity, and how the activity of beta 1 integrin can be modulated by activators of cell adhesion. PMID- 7499397 TI - A conserved glutamate is responsible for ion selectivity and pH dependence of the mammalian anion exchangers AE1 and AE2. AB - The erythrocyte anion exchanger AE1 (band 3) serves as an important model for the study of the mechanism of ion transport. Chemical modification of human erythrocyte AE1 has previously suggested that glutamic acid residue 681 lies within the transport pathway and can cross the permeability barrier. This glutamate is conserved in all anion exchangers sequenced to date. We examined the effect on divalent (sulfate) and monovalent (chloride and bicarbonate) anion transport of mutating the corresponding glutamates in mouse AE1 and the closely related anion exchanger, AE2. Substitution of this conserved glutamate with uncharged or basic amino acids had a negligible effect on the maximal rate of sulfate-sulfate exchange in AE-reconstituted proteoliposomes, but largely abolished the steep pH dependence of sulfate transport observed in wild-type AE1 and AE2. In contrast, exchange of monovalent anions was undetectable in cells expressing these mutants. Replacement of the conserved glutamate with aspartate abolished both monovalent and divalent anion transport. These data suggest that the conserved glutamate residue plays a dual role in determining anion selectivity and in proton coupling to sulfate transport. A model explaining the role of the conserved glutamate in promoting ion selectivity and pH regulation is discussed. PMID- 7499398 TI - Pressure-induced dissociation and denaturation of allophycocyanin at subzero temperatures. AB - The thermodynamics of assembly of the allophycocyanin hexamer was examined employing hydrostatic pressures in the range of 1 bar to 2.4 kbar and temperatures of 20 to -12 degrees C, the latter made possible by the decrease of the freezing point of water under pressure. The existence of two processes, dissociation of the hexamer into dimers, (alpha beta)3-->3 (alpha beta), and dissociation of the alpha beta dimers into monomers, (alpha beta)-->alpha + beta have been recognized previously by changes in the absorbance and fluorescence of the tetrapyrrolic chromophores owing to added ligands. The same changes are observed in the absence of ligands at pressures of under 2.4 kbar and temperatures down to -12 degrees C. On decompression from 2.4 kbar at 0 degrees C, appreciable hysteresis and a persistent loss of 50% in the absorbance at 653 nm is observed. It results from the conformational drift of the isolated subunits and is reduced to 10% when the highest pressure is limited to 1.6 kbar. The thermodynamic parameters of the reaction alpha + beta-->alpha beta can be determined from pressure effects on perchlorate solutions of allophycocyanin, which consist of dimers alone. Their previous knowledge permits estimation, under suitable hypotheses, of the thermodynamic parameters of the reaction 3(alpha beta)-->(alpha beta)3 from the overall pressure effects on the hexamers. Both association reactions have positive enthalpy changes, and the whole hexamer assembly is made possible by the excess entropy. PMID- 7499399 TI - Intracellular localization and metabolism of chylomicron remnants in the livers of low density lipoprotein receptor-deficient mice and apoE-deficient mice. Evidence for slow metabolism via an alternative apoE-dependent pathway. AB - The metabolism of chylomicron remnants in mice deficient in low density lipoprotein receptor (LDLr) or apolipoprotein E (apoE) was compared with that of control C57BL/6J mice. Mice were injected intravenously with chylomicron-like emulsions labeled with radioactive lipids. Blood samples were taken at fixed time intervals from the retro-orbital sinus, and clearance rates of the lipoproteins were assessed from the decline in plasma radioactivities. To follow the intracellular pathway of remnants in the liver, emulsions labeled with a fluorescent cholesteryl ester (BODIPY) were injected, and liver sections were processed and assayed by laser confocal microscopy. Catabolism of remnant cholesteryl esters was assessed by injecting emulsions labeled with cholesteryl[1 14C]oleate and measuring the expired CO2 from each animal. In apoE-deficient mice, remnant removal from plasma was totally impeded, while the clearance of remnants in LDLr-deficient mice was similar to that in C57BL/6J control mice. The confocal micrographs of livers 20 min after injection of fluorescent chylomicron like emulsions showed evenly distributed fluorescent particles in the hepatocytes from control mice. In contrast, the fluorescent particles were mainly located in sinusoidal spaces in LDLr-deficient mice. Three hours after injection the livers from control mice showed few fluorescent particles, indicating that remnants have been catabolized, while the sections from LDLr-deficient mice were still highly fluorescent. Micrographs from apoE-deficient mice showed no fluorescent particles in the liver at any time after injection. Measurement of expired radioactive CO2 after injection of emulsions labeled in the fatty acid moiety of cholesteryl oleate indicated that remnant metabolism was slower in the LDLr-deficient mice and essentially nil in the apoE-deficient mice. Control mice had expired 50% of the injected label by 3 h after injection. We conclude that under normal circumstances, chylomicron remnants are rapidly internalized by LDLr and catabolized in hepatocytes, with a critical requirement for apoE. When LDLr is absent, remnants are taken up by a second apoE-dependent pathway, first to the sinusoidal space of the liver, with subsequent slow endocytosis and slow catabolism. Hepatic clearance via this second pathway is increased by heparin, inhibited by lactoferrin, heparinase, and suramin, and down-regulated by feeding a high fat diet. PMID- 7499400 TI - The cardiac inward rectifier K+ channel subunit, CIR, does not comprise the ATP sensitive K+ channel, IKATP. AB - Cardiac IKACh is comprised of two inwardly rectifying K+ channel subunits, CIR and GIRK1 (Krapivinsky, G., Gordon, E. G., Wickman, K., Velimirovic, B., Krapivinsky, L., and Clapham, D. E. (1995) Nature 374, 135-141). A cardiac protein virtually identical to CIR, termed rcKATP-1 (Ashford, M. L. J., Bond, C. T., Blair, T. A., and Adelman, J. P. (1994) Nature 370, 456-459), was reported to form an ATP-sensitive inwardly rectifying K+ channel, IKATP. We attempted to determine whether CIR alone or together with an unknown protein(s) participated in the formation of cardiac IKATP. Expression of CIR in insect, oocyte, and mammalian cell systems did not increase the appearance of ATP-sensitive currents, but rather gave rise to unique strongly inwardly rectifying, G protein-regulated K+ currents. CIR protein is found exclusively in atria, in contrast to the predominance of IKATP functional activity in ventricle. Also, CIR was completely depleted from heart membrane after immunodepletion of GIRK1. We conclude that CIR/rcKATP-1 is not a subunit of cardiac IKATP and that GIRK1 is the only channel protein coassociating with CIR in heart. PMID- 7499401 TI - Heterogeneous nuclear ribonucleoproteins H, H', and F are members of a ubiquitously expressed subfamily of related but distinct proteins encoded by genes mapping to different chromosomes. AB - Molecular cDNA cloning, two-dimensional gel immunoblotting, and amino acid microsequencing identified three sequence-unique and distinct proteins that constitute a subfamily of ubiquitously expressed heterogeneous nuclear ribonucleoproteins corresponding to hnRNPs H, H', and F. These proteins share epitopes and sequence identity with two other proteins, isoelectric focusing sample spot numbers 2222 (37.6 kDa; pI 6.5) and 2326 (39.5 kDa; pI 6.6), indicating that the subfamily may contain additional members. The identity between hnRNPs H and H' is 96%, between H and F 78%, and between H' and F 75%, respectively. The three proteins contain three repeats, which we denote quasi RRMs (qRRMs) since they have a remote similarity to the RNA recognition motif (RRM). The three qRRMs of hnRNP H, with a few additional NH2-terminal amino acids, were constructed by polymerase chain reaction amplification and used for ribohomopolymer binding studies. Each qRRM repeat bound poly(rG), while only the NH2-terminal qRRM bound poly(rC) and poly(rU). None of the repeats bound detectable amounts of poly(rA). The expression levels of hnRNPs H and F were differentially regulated in pairs of normal and transformed fibroblasts and keratinocytes. In normal human keratinocytes, the expression level of H was unaffected by treatment with several substances tested including two second messengers and seven cytokines. Likewise the expression level of F was independent of these substances, although it was strikingly down-regulated by long term treatment with 4 beta-phorbol 12-myristate 13-acetate, indicating that the protein kinase C signaling pathway regulates its expression. No effect of 4 beta-phorbol 12-myristate 13-acetate was observed on the expression of hnRNP H. The genes coding for hnRNPs H, H', and F were chromosome-mapped to 5q35.3 (HNRPH1), 6q25.3-q26, and/or Xq22 (HNRPH2) and 10q11.21-q11.22 (HNRPF), respectively. PMID- 7499402 TI - Effect of ionizing radiation on AP-1 binding activity and basic fibroblast growth factor gene expression in drug-sensitive human breast carcinoma MCF-7 and multidrug-resistant MCF-7/ADR cells. AB - We studied the effect of ionizing radiation on the activation of the AP-1 transcription factors and the regulation of basic fibroblast growth factor (bFGF) gene expression in drug-sensitive human breast carcinoma (MCF-7) cells and its drug-resistant variant (MCF-7/ADR) cells. Northern blot and gel mobility shift assays showed that 135 cGy of ionizing radiation induced c-jun and c-fos gene expression, AP-1 binding activity, as well as bFGF gene expression in MCF-7/ADR cells. In MCF-7 cells, however, we observed little/no induction of bFGF gene expression and AP-1 binding activity after the stress. Nevertheless, MCF-7 cells transfected with plasmids containing c-jun gene contain high levels of bFGF protein. H-7 (60 micrograms/ml), a potent protein kinase C (PKC) inhibitor, inhibited the stress-induced AP-1 binding activity and bFGF gene expression in MCF-7/ADR cells. Corroborating this observation, overexpression of PKC alpha induced bFGF gene expression in MCF-7 cells. Taken together, these results suggest that stress-induced bFGF gene expression is mediated through the activation of PKC and AP-1 transcription factors. Differences in the levels of PKC activity and AP-1 binding factors may be responsible for differential expression of bFGF among breast cancer cell lines. Although there are large differences in response to ionizing radiation between MCF-7 and MCF-7/ADR cell lines, we observed no significant differences in radiocytotoxicity between them. PMID- 7499403 TI - Induction of apoptosis by the c-Myc helix-loop-helix/leucine zipper domain in mouse 3T3-L1 fibroblasts. AB - The cellular proto-oncogene c-myc is involved in cell proliferation and transformation but is also implicated in the induction of programmed cell death (apoptosis). The c-Myc protein is a transcriptional activator with a carboxyl terminal basic region/helix-loop-helix (HLH)/leucine zipper (LZ) domain. It forms heterodimers with the HLH/LZ protein Max and transactivates gene expression after binding DNA E-box elements. We have studied the phenotype of dominant-negative mutants of c-Myc and Max in microinjection experiments. Max mutants with a deleted or mutated basic region inhibited DNA synthesis in serum-stimulated 3T3 L1 mouse fibroblasts. In contrast, mutants of c-Myc expressing only the basic region/HLH/LZ or HLH/LZ domains rapidly induced apoptosis at low and high serum levels. Co-expression of the HLH/LZ domains of c-Myc and Max failed to do so. We suggest that the c-Myc HLH/LZ domain induces apoptosis by specific interaction with cellular factors different to Max. PMID- 7499404 TI - Affinity chromatography demonstrates a direct binding between cytoplasmic dynein and the dynactin complex. AB - We used affinity chromatography to probe for a direct binding interaction between cytoplasmic dynein and dynactin. Purified cytoplasmic dynein was found to bind to an affinity column of p150Glued, the largest polypeptide in the dynactin complex. To test the specificity of the interaction, we loaded rat brain cytosol onto the p150Glued affinity column and observed that cytoplasmic dynein from cytosol was specifically retained on the column. Preincubation of the p150Glued affinity matrix with excess exogenous dynein intermediate chain resulted in a significant reduction of dynein binding, suggesting that p150Glued may be interacting with dynein via this polypeptide. Therefore we constructed an affinity column of recombinant dynein intermediate chain and observed that dynactin was retained from rat brain cytosol. These results demonstrate that the native dynein and dynactin complexes are capable of direct in vitro interaction mediated by a direct binding of the dynein intermediate chain to the p150Glued component of the dynactin complex. We have mapped the site of this interaction to the amino terminal region of p150Glued, which is predicted to form an alpha-helical coiled coil. Regulation of the dynein-dynactin interaction may prove to be key in the control mechanism for cytoplasmic dynein-mediated vesicular transport. PMID- 7499405 TI - Interaction of integrin alpha IIb beta 3 with multiple fibrinogen domains during platelet adhesion. AB - We have investigated how modulation of integrin alpha IIb beta 3 function influences the mechanisms that initiate platelet thrombus formation onto surface bound fibrinogen and isolated fibrinogen domains. Under stationary conditions and with full activation of platelets blocked by prostaglandin E1, the carboxyl terminal gamma 400-411 sequence is necessary for establishing initial contact with the immobilized substrate. Molecules containing a single copy of this sequence, like the plasmin-generated fibrinogen fragment D, support platelet spreading, but the resulting attachment to the surface is loose and disrupted by minimal peeling force. In contrast, platelets adhere firmly to intact fibrinogen under the same conditions, suggesting that recognition of contact sites outside a single D domain can secure the firm interaction not supported by a single gamma 400-411 sequence. If platelets are activated, the gamma 400-411 sequence is no longer necessary to initiate the adhesion process but becomes sufficient, even as a single copy, to mediate stable surface attachment in the absence of shear stress. Under conditions of flow, however, intact fibrinogen but not fragment D can support adhesion, regardless of whether platelets have the potential to become activated or not. These results indicate the functional relevance of multiple fibrinogen domains during the initial stages of the platelet adhesion process. PMID- 7499406 TI - Conformational cycle of the archaeosome, a TCP1-like chaperonin from Sulfolobus shibatae. AB - The major heat shock proteins in the archaeon Sulfolobus shibatae are similar to the cytosolic eukaryotic chaperonin and form an 18-subunit bitoroidal complex. Two sequence-related subunits constitute a functional complex, named the archaeosome. The archaeosome exists in two distinct conformational states that are part of chaperonin functional cycle. The closed archaeosome complex binds ATP and forms an open complex. Upon ATP hydrolysis, the open complex dissociates into subunits. Free subunits reassemble into a two-ring structure. The equilibrium between the complexes and free subunits is affected by ATP and temperature. Denatured proteins associate with both conformational states as well as with free subunits that form an intermediate complex. These unexpected observations suggest a new mechanism of archaeosome-mediated thermotolerance and protein folding. PMID- 7499408 TI - Inhibition of Ras/Raf interaction by anti-oncogenic mutants of neurofibromin, the neurofibromatosis type 1 (NF1) gene product, in cell-free systems. AB - The neurofibromatosis type 1 (NF1) gene encodes a protein, neurofibromin, containing GTPase-activating protein-related domain (GRD) that stimulates intrinsic GTPase activity of Ras protein. By screening a randomly mutagenized NF1 GRD library in Saccharomyces cerevisiae, we isolated two NF1-GRD mutants (NF201 and NF204) with single amino acid substitutions, which suppress the heat shock sensitive phenotype of the RAS2(G19V) mutant. The NF1-GRD mutants also suppress the oncogenic Ras-induced transformation of NIH 3T3 mouse fibroblasts (Nakafuku, M., Nagamine, M., Ohtoshi, A., Tanaka, K., Toh-e, A., and Kaziro, Y. (1993) Proc. Natl. Acad. Sci. U.S.A. 90, 6706-6710). In this paper, we investigated the molecular mechanism of inhibition of the transforming Ras-specific function by the NF1-GRD mutants in mammalian cells. In human embryonic kidney (HEK) 293 cells, the mutant NF1-GRDs attenuated the stimulation of mitogen-activated protein kinase by Ras(G12V), but not by platelet-derived growth factor. In cell free systems, purified recombinant NF1-GRD mutants showed an inhibitory effect on the association of Ras.guanosine 5'-O-(3-thiotriphosphate) (GTP gamma S) with Raf at several times lower concentrations than the wild type. Furthermore, it was revealed that the binding affinity of the mutant NF1-GRDs toward Ras.GTP gamma S is approximately 5-10 times higher than the wild type. These results suggest that the mutant NF1-GRDs tightly bind to an oncogenic Ras in its GTP-bound active conformation and block the interaction between Ras and its effector, Raf. PMID- 7499407 TI - Epithelial membrane protein-1, peripheral myelin protein 22, and lens membrane protein 20 define a novel gene family. AB - Peripheral myelin protein 22 (PMP22) is expressed in many tissues but mainly by Schwann cells as a component of compact myelin of the peripheral nervous system (PNS). Mutations affecting PMP22 are associated with hereditary motor and sensory neuropathies. Although these phenotypes are restricted to the PNS, PMP22 is thought to play a dual role in myelin formation and in cell proliferation. We describe the cloning and characterization of epithelial membrane protein-1 (EMP 1), a putative four-transmembrane protein of 160 amino acids with 40% amino acid identity to PMP22. EMP-1 and PMP22 are co-expressed in most tissues but with differences in relative expression levels. EMP-1 is most prominently found in the gastrointestinal tract, skin, lung, and brain but not in liver. In the corpus gastricum, EMP-1 protein can be detected in epithelial cells of the gastric pit and isthmus of the gastric gland in a pattern consistent with plasma membrane association. EMP-1 and PMP22 mRNA levels are inversely regulated in the degenerating rat sciatic nerve after injury and by growth arrest in NIH 3T3 fibroblasts. The discovery of EMP-1 as the second member of a novel gene family led to the identification of the lens-specific membrane protein 20 (MP20) as a third but distant relative. The proteins of this family are likely to serve similar functions possibly related to cell proliferation and differentiation in a variety of cell types. PMID- 7499409 TI - Cloning, expression, and characterization of the TATA-binding protein (TBP) promoter binding factor, a transcription activator of the Acanthamoeba TBP gene. AB - TATA-binding protein (TBP) gene promoter binding factor (TPBF) is a transactivator which binds to the TBP promoter element (TPE) sequence of the Acanthamoeba TBP gene promoter and stimulates transcription in vitro. We have isolated a cDNA clone encoding TPBF. TPBF is a polypeptide of 327 amino acids with a calculated molecular mass of 37 kDa. The predicted amino acid sequence of TPBF shows no significant homology to other proteins. TPBF has two potential coiled-coil regions, a basic region, a proline-rich region, a histidine-rich N terminus, and a nuclear targeting sequence. The recombinant protein has an apparent molecular mass of 50 kDa, identical with that of TPBF purified from Acanthamoeba. Recombinant TPBF is able to bind DNA and activate transcription with the same specificity as natural Acanthamoeba TPBF, demonstrating the authenticity of the clone. Mobility shift assays of co-translated TPBF polypeptides and chemical cross-linking demonstrate that TPBF is tetrameric in solution and when bound to DNA. Analyses of TPBF mutants show that Coiled-coil II is essential for DNA binding, but Coiled-coil I and the basic region are also involved. TPBF is thus a novel DNA-binding protein with functional similarity to the tumor suppressor protein p53. PMID- 7499410 TI - Transcription of cystic fibrosis transmembrane conductance regulator requires a CCAAT-like element for both basal and cAMP-mediated regulation. AB - The cystic fibrosis transmembrane conductance regulator (CFTR) gene in man is controlled by a tightly regulated and weak promoter. The architecture of the CFTR promoter suggests regulatory characteristics that are consistent with the absence of a TATA-like sequence, including the ability to initiate RNA transcription at numerous positions. Detailed investigation of the most proximal region of the human CFTR gene promoter through deletion and mutational analysis reveals that expression is contingent on the conservation of the inverted CCAAT sequence. Basal expression of CFTR transcription and cAMP-mediated transcriptional regulation require the presence of an imperfect and inverted CCAAT element recognized as 5'-AATTGGAAGCAAAT-3', located between 132 and 119 nucleotides upstream of the translational start site. RNA isolated from a transfected pancreatic cell line carrying integrated wild-type and mutant CFTR-directed transgenes was used to map the 5' termini of the transgenic transcripts. Analysis of the transcript termini by ribonuclease protection analysis reflects the direct association of the conserved inverted CCAAT sequence in promoting transcript initiation. Because of the requirement for the inverted CCAAT sequence for promoting transcription of CFTR, the involvement of CCAAT-binding factors is suspected in the regulation of CFTR gene transcription. To test this, we used electrophoretic mobility shift assays to demonstrate that the majority of the binding to the inverted CCAAT element, between -135 and -116, was easily competed for by binding to cognate nucleotide sequences for CCAAT-enhancer binding protein (C/EBP). An antibody specific for the C/EBP-related protein, C/EBP delta, detected C/EBP delta as part of a nuclear protein complex bound to the inverted CCAAT sequence of the CFTR gene. Also, the detection of specific activating transcription factor/cyclic-AMP response element binding protein antigens by antibody supershift analysis of nuclear complexes suggest that species of this family of transcription factors could be involved in the formation of complexes with C/EBP delta within the CFTR gene inverted CCAAT-like element. These studies raise the possibility of interactions between individual members of the C/EBP and activating transcription factor/cyclic-AMP response element binding protein families potentially contribute to the tight transcriptional control rendered by the CFTR gene promoter. PMID- 7499411 TI - Analysis of interleukin-2-dependent signal transduction through the Shc/Grb2 adapter pathway. Interleukin-2-dependent mitogenesis does not require Shc phosphorylation or receptor association. AB - The interleukin (IL)-2 receptor system has previously been shown to signal through the association and tyrosine phosphorylation of Shc. This study demonstrates that the IL-2 receptor beta (IL-2R beta) chain is the critical receptor component required to mediate this effect. The use of IL-2R beta chain deletion mutants transfected into a Ba/F3 murine cell model describes a requirement for the IL-2R beta "acid-rich" domain between amino acids 315 and 384 for Shc tyrosine phosphorylation and receptor association. COS cell co transfection studies of IL-2R beta chain constructs containing point mutations of tyrosine to phenylalanine along with the tyrosine kinase Jak-1 and a hemagglutinin-tagged Shc revealed that the motif surrounding phosphorylated tyrosine 338 within the acid-rich domain of the IL-2R beta is a binding site for Shc. Deletion of this domain has previously been shown to abrogate the ability of IL-2 to activate Ras but does not affect IL-2-dependent mitogenesis in the presence of serum. Proliferation assays of Ba/F3 cells containing IL-2R beta chain deletion mutants in serum-free medium with or without insulin shows that deletion of the acid-rich domain does not affect IL-2-driven mitogenesis regardless of the culture conditions. This study thus defines the critical domain within the IL-2R beta chain required to mediate Shc binding and Shc tyrosine phosphorylation and further shows that Shc binding and phosphorylation are not required for IL-2-dependent mitogenesis. Neither serum nor insulin is required to supplement the loss of induction of the Shc adapter or Ras pathways, which therefore suggests a novel mechanism for mitogenic signal transduction mediated by this hematopoietin receptor. PMID- 7499412 TI - Identification of active site residues of the Tsp protease. AB - In a search for active-site residues of the Tsp protease, 20 positions were individually mutated to alanine, the mutant strains were assayed for growth defects in vivo, and the purified proteins were assayed for proteolytic activity in vitro. Alanine substitutions at three positions, Ser-430, Asp-441, and Lys 455, result in inactive proteases that have structures and substrate-binding properties similar to wild type, suggesting that the side chains at these positions participate in catalysis. Replacing Ser-430 with cysteine results in a partially active protease, which is inhibited by cysteine-modifying reagents. Replacing Asp-441 with asparagine does not significantly affect activity. However, other residues, including histidine and arginine, cannot functionally replace Lys-455. These data are consistent with a serine-lysine dyad mechanism, similar to those proposed for the LexA-like proteases, the type I signal peptidases, and the class A beta-lactamases. PMID- 7499413 TI - Identification of the binding domain for secretory phospholipases A2 on their M type 180-kDa membrane receptor. AB - The rabbit muscle (M)-type receptor for secretory phospholipases A2 (sPLA2s) has a large extracellular domain of 1394 amino acids, composed of an N-terminal cysteine-rich domain, a fibronectin-like type II domain, and eight carbohydrate recognition domains (CRDs). It is thought to mediate some of the physiological effects of mammalian sPLA2s, including vascular smooth muscle contraction and cell proliferation, and is able to internalize sPLA2s. Here, we show by site directed mutagenesis that OS1, a snake venom sPLA2, binds to the receptor via its CRDs and that deletion of CRD 5 completely abolishes the binding of sPLA2s. Moreover, a receptor lacking all CRDs but CRD 5 was still able to bind OS1 although with a lower affinity. Deletion of CRDs 4 and 6, surrounding the CRD 5, slightly reduced the affinity for OS1, thus suggesting that these CRDs are also involved in the binding of OS1. The M-type sPLA2 receptor and the macrophage mannose receptor are homologous and are predicted to share the same tertiary structure. p-Aminophenyl-alpha-D-mannopyranoside bovine serum albumin, a known ligand of the macrophage mannose receptor, binds to the M-type sPLA2 receptor essentially via CRDs 3-6. PMID- 7499414 TI - Purification of a galactosyl-alpha 1-4-galactose-binding adhesin from the gram positive meningitis-associated bacterium Streptococcus suis. AB - Streptococcus suis causes meningitis, sepsis, and other serious infections in newborn and young pigs and in adult humans. The Gal alpha 1-4Gal-binding adhesin of S. suis was purified to homogeneity by ultrasonic treatment, fractional ammonium sulfate precipitation, and preparative polyacrylamide gel electrophoresis. Pigeon ovomucoid, a glycoprotein with Gal alpha 1-4Gal terminals, was used to detect the adhesin by blotting. The purified adhesin appeared as single band of an apparent size of 18 kDa and of a pI of 6.4; no disulfide bridges were present. The amount of adhesin as revealed by pigeon ovomucoid binding correlated with the hemagglutination activity of different S. suis strains. The purified adhesin bound to latex particles induced hemagglutination which was specifically inhibited with the same inhibitors as hemagglutination by the intact bacteria, thus demonstrating that the purified protein was the Gal alpha 1-4Gal-recognizing adhesin of S. suis. Two adhesin variants (PN and PO) with differing Gal alpha 1-4Gal binding specificity had the similar electrophoretic mobilities and the same N-terminal peptide sequences, indicating that they were closely related. This represents the first isolation of an adhesin with well-defined cell surface carbohydrate binding activity from Gram positive bacteria associated with meningitis. PMID- 7499415 TI - Brefeldin A reversibly inhibits the assembly of apoB containing lipoproteins in McA-RH7777 cells. AB - BFA inhibited in a dose dependent way the assembly of apoB-48 very low density lipoprotein (VLDL) but allowed a normal rate of biosynthesis of the apolipoprotein and of the assembly of the dense ("high density lipoprotein (HDL) like") apoB-48 particle (apoB-48 HDL). The inhibition of the assembly of apoB-48 VLDL occurred at BFA levels that allowed a major secretion of both transferrin and apoB-48 HDL. The assembly of apoB-100 containing lipoproteins was also inhibited by BFA but could be reactivated by a 30-60 min chase in the absence of BFA, which agreed with the time that was estimated to be needed to restore the secretory pathway (approximately 60 min). Also the assembly of apoB-48 VLDL was reversible. Both apoB-48 and apoB-100 that was labeled in the presence of BFA assembled VLDL after removal of the BFA. Both apoB-100 and apoB-48 were associated with the membrane pellet of the microsomes. Virtually all (122 +/- 30%) of the membrane associated pulse-labeled apoB-48 remained in the membrane after a 180-min chase in the presence of BFA, compared to only 21 +/- 2% in normal cells (mean +/- S.D., n = 4). The corresponding figures for apoB-100 was 40 +/- 7% in BFA-treated cells and 9 +/- 7% in normal cells (mean +/- S.D., n = 4). Pulse-chase experiments with BFA offered conditions to selectively follow the turnover of membrane-associated apoB-100. Such experiments indicated that this apoB-100 pool is a precursor to VLDL. PMID- 7499417 TI - In HepG2 cells, translocation, not degradation, determines the fate of the de novo synthesized apolipoprotein B. AB - Previous studies show that translocation and degradation of apolipoprotein B (apoB), two processes occurring on or within the endoplasmic reticulum, determine how much de novo synthesized apoB is secreted. We determined which of these processes regulates the intracellular fate of apoB by examining whether degradation determines how much apoB is translocated or if translocation determines how much apoB is degraded. HepG2 cells, treated with the cysteine active site protease inhibitor ALLN, previously shown to block the degradation of translocation-arrested apoB in Chinese hamster ovary cells (Du, E., Kurth, J., Wang, S.-L., Humiston, P., and Davis, R.A. (1994) J. Biol. Chem. 269, 24169 24176), showed a 10-fold increase in the accumulation of de novo synthesized [35S]methionine-labeled apoB. The majority (80%) of the apoB accumulated in response to ALLN was in the microsomal fraction. In contrast, ALLN did not effect apoB secretion. Since ALLN did not effect the intracellular accumulation of [35S]methionine-labeled albumin and other proteins (trichloroacetic acid precipitable [35S]methionine-labeled proteins), its effect on apoB was specific. Pulse-chase studies showed that ALLN dramatically reduced the first-order rate of removal of [35S]methionine-labeled apoB from the cell but did not effect its rate of secretion. The finding that ALLN caused the intracellular accumulation of incompletely translated chains of apoB suggests that at least some of the degradation occurs at the ribosomal level. Moreover, 85% of the apoB that accumulated in isolated microsomes in response to ALLN was accessible to exogenous trypsin, indicating this pool of apoB was incompletely translocated. The combined data suggest that translocation, not degradation, determines the intracellular fate of de novo synthesized apoB. PMID- 7499416 TI - The mouse obese gene. Genomic organization, promoter activity, and activation by CCAAT/enhancer-binding protein alpha. AB - The obese gene product, leptin, regulates adiposity. Mice homozygous for a nonfunctional obese gene become massively obese and develop diabetes mellitus due to overeating and increased metabolic efficiency. The cDNA sequence of obese was recently reported (Zhang, Y., Proenca, R., Maffei, M., Barone, M., Leopold, L., and Friedman, J. L. (1994) Nature 372, 425-432; Correction: (1995 Nature 374, 479). We have determined the genomic organization of the 5' end of the mouse obese gene. The coding sequence is in exons 2 and 3. A single TATA-containing promoter was found upstream of exon 1. A minority (probably approximately 5%) of the obese mRNA contained an extra, untranslated exon between exons 1 and 2. Transcription of the obese gene was detected only in adipose cells. A 762-base pair obese gene promoter driving a luciferase gene yielded abundant activity in transiently transfected rat adipose cells in primary culture. The obese promoter was inactive in erythroid K562 cells. Deletion of bases from -762 downstream to 161 did not affect promoter activity in transfected adipose cells. The -161 minimal promoter contained consensus Sp1 and CCAAT/enhancer-binding protein (C/EBP) motifs. Cotransfection with C/EBP alpha (a transcription factor important in adipose cell differentiation) caused 23-fold activation. These data suggest that the obese promoter is a natural target of C/EBP alpha. PMID- 7499418 TI - Isolation of MEK5 and differential expression of alternatively spliced forms. AB - The prototype mitogen-activated protein (MAP) kinase module is a three-kinase cascade consisting of the MAP kinase, extracellular signal-regulated protein kinase (ERK) 1 or ERK2, the MAP/ERK kinase (MEK) MEK1 or MEK2, and the MEK kinase, Raf-1 or B-Raf. This and other MAP kinase modules are thought to be critical signal transducers in major cellular events including proliferation, differentiation, and stress responses. To identify novel mammalian MAP kinase modules, polymerase chain reaction was used to isolate a new MEK family member, MEK5, from the rat. MEK5 is more closely related to MEK1 and MEK2 than to the other known mammalian MEKs, MKK3 and MKK4. MEK5 is thought to lie in an uncharacterized MAP kinase pathway, because MEK5 does not phosphorylate the ERK/MAP kinase family members ERK1, ERK2, ERK3, JNK/SAPK, or p38/HOG1, nor will Raf-1, c-Mos, or MEKK1 highly phosphorylate it. Alternative splicing results in a 50-kDa alpha and a 40-kDa beta isoform of MEK5. MEK5 beta is ubiquitously distributed and primarily cytosolic. MEK5 alpha is expressed most highly in liver and brain and is particulate. The 23 amino acids encoded by the 5' exon in the larger alpha isoform are similar to a sequence found in certain proteins believed to associate with the actin cytoskeleton; this alternatively spliced modular domain may lead to the differential subcellular localization of MEK5 alpha. PMID- 7499420 TI - Identification and characterization of a novel squamous cell-associated gene related to PMP22. AB - In this study, we identify and characterize a novel gene, CL-20, that encodes a 17.8-kDa protein with sequence and structural similarity to the growth arrest specific gene gas3/peripheral myelin protein gene PMP22. The CL-20 protein exhibits a 43% identity with PMP22. The positions of the four lipophilic domains and the N-glycosylation site of PMP22 are conserved in CL-20, suggesting that it also is an integral membrane glycoprotein. The CL-20 gene is located on human chromosome 12 rather than 17 and encodes a 2.8-kilobase mRNA instead of 1.7 kilobase mRNA. These observations indicate that the CL-20 gene is related to but distinct from PMP22. In contrast to PMP22, CL-20 mRNA and protein are induced during squamous differentiation of rabbit tracheal epithelial cells in vitro, and Northern blot analysis and in situ hybridization demonstrated that CL-20 mRNA is most abundant in squamous epithelia. These results indicate that the high expression of CL-20 is closely correlated with squamous differentiation. The differences in tissue-specific expression and regulation between CL-20 and PMP22 suggest different roles for these two proteins. Retinoids, which inhibit squamous differentiation, repress the induction of CL-20. The retinoic acid receptor selective retinoid SRI-6751-84 is the most effective in suppressing CL-20, suggesting that the activation of the retinoic acid receptor signaling pathway is important in this suppression. PMID- 7499419 TI - Sp1 is a component of the cytokine-inducible enhancer in the promoter of vascular cell adhesion molecule-1. AB - Transcription of the vascular cell adhesion molecule-1 (VCAM-1) gene in endothelial cells is induced by the inflammatory cytokines interleukin-1 beta, tumor necrosis factor-alpha, and lipopolysaccharide. Previous studies demonstrated that the cytokine-response region in the VCAM1 promoter contains binding sites for the transcription factors nuclear factor-kappa B (NF-kappa B) and interferon regulatory factor-1. Using a saturation mutagenesis approach, we report that the cytokine-inducible enhancer consists of these previously characterized elements and a novel region located 3' of the NF-kappa B sites. Electrophoretic mobility shift assays and DNase I footprint studies with endothelial nuclear extracts and recombinant protein revealed that the transcriptional activator Sp1 interacts with this novel element in a specific manner. Transient transfection assays using vascular endothelial cells revealed that site-directed mutations in the Sp1 binding element decreased tumor necrosis factor-alpha-induced activity of the VCAM1 promoter. The cytokine-induced enhancer of the VCAM1 gene requires constitutively bound Sp1 and induced heterodimeric NF-kappa B for maximal promoter activity. PMID- 7499421 TI - A novel cis-acting element in a liver cytochrome P450 3A gene confers synergistic induction by glucocorticoids plus antiglucocorticoids. AB - The induction by dexamethasone of rat liver CYP3A1 differs from classical glucocorticoid gene regulation in part because both glucocorticoids and antiglucocorticoids such as pregnenolone 16 alpha-carbonitrile (PCN) induce CYP3A1 through transcriptional gene activation. In the present study, we transiently expressed in primary cultures of rat hepatocytes plasmids consisting of CYP3A1 5'-flanking sequences fused to a chloramphenicol acetyltransferase reporter plasmid. Deletional analysis identified a 78-base pair (bp) element located approximately 135 bp upstream of the transcriptional start site that was inducible by treatment of the cultures with dexamethasone or PCN and was induced synergistically by dexamethasone plus PCN. Nuclear extract from control rat liver protected two regions within the 78-bp sequence against digestion with DNase I. The same two regions were protected when nuclear extracts from dexamethasone treated animals were used. Analysis of both of the "footprints" (FP1 and FP2) failed to reveal a classical sequence for the glucocorticoid-responsive element. A 33-bp element that includes FP1 sequences inserted into the chloramphenicol acetyltransferase reporter plasmid and transiently expressed in rat hepatocytes conferred a profile of dexamethasone and PCN induction similar to that of the 78 bp element. However, an Escherichia coli expressed glucocorticoid receptor protein failed to protect sequences within FP1 in DNase I footprinting experiments and failed to change its mobility in gel shift assays. Moreover, as judged by the gel shift assay, the specific protein binding to this fragment was the same whether nuclear extracts from the liver of untreated or dexamethasone treated rats were used. We conclude that the activation of CYP3A1 gene transcription by glucocorticoids may involve proteins already bound to the controlling element in the CYP3A1 gene through a mechanism in which GR in the presence of hormone does not bind directly to CYP3A1 DNA. PMID- 7499423 TI - Subunit structure of mitochondrial DNA polymerase from Drosophila embryos. Physical and immunological studies. AB - The subunit structure of mitochondrial DNA polymerase from Drosophila embryos has been examined by a combination of physical and immunological methods. A highly specific rabbit antiserum directed against the native enzyme was developed and found to recognize specifically its two subunits in immunoblot and immunoprecipitation analyses. That and the potent inhibition by the rabbit antiserum of the DNA polymerase and 3'-->5' exonuclease activities of the nearly homogeneous mitochondrial DNA polymerase provide strong evidence for the physical association of the 3'-->5' exonuclease with the two subunit enzyme. An immunoprecipitation analysis of crude enzyme fractions showed that the two subunits of Drosophila mitochondrial DNA polymerase are intact, and an in situ gel proteolysis analysis showed that they are structurally distinct. Template primer DNA binding studies demonstrated formation of a stable and discrete enzyme DNA complex in the absence of accessory proteins. Photochemical cross-linking of the complexes by UV light indicated that the alpha but not the beta subunit of mitochondrial DNA polymerase makes close contact with DNA, and limited digestion of the native enzyme with trypsin showed that an approximately 65-kDa proteolytic fragment of the alpha subunit retains the DNA binding function. PMID- 7499422 TI - Src homologous and collagen (Shc) protein binds to F-actin and translocates to the cytoskeleton upon nerve growth factor stimulation in PC12 cells. AB - Immunoprecipitates of metabolically labeled PC12 cells consistently contained a 43-kDa protein that was associated with Shc, a signal-transducing protein with a single SH2 domain. Following affinity chromatography with immobilized recombinant glutathione S-transferase (GST)-Shc fusion protein, the 43-kDa protein was identified as actin by mass spectrometry and immunoblotting. Cosedimentation experiments using purified actin and GST-Shc showed that Shc binds directly to F actin, confirming Shc-actin interaction in vivo. Various GST-truncated Shc fusion proteins were prepared and used in actin cosedimentation assays. Constructs containing the SH2 and collagen homology domains were not precipitated, and those containing the amino-terminal domain were. Thus, Shc-actin interactions do not occur in the region of tyrosine phosphorylation and leave the SH2 domain free to bind to other tyrosine-phosphorylated molecules. Although the major pool of Shc in unstimulated PC12 cells is soluble, two other pools are associated with the cytoskeleton and the submembranous cytoskeleton. Upon nerve growth factor stimulation, approximately 50% of the soluble Shc translocates to both cytoskeleton environments within 2 min, decreasing thereafter. When cells were pretreated with cytochalasin D, a drug that disrupts actin filaments, Shc translocation to the cytoskeleton was abolished. However, in the submembranous fraction, the Shc level was elevated in resting cells following cytochalasin D treatment. The kinetics of translocation, compared to mitogen-activated protein kinase activation, and the nature of the Shc-actin interaction suggest that the cytoskeletal association of Shc, induced by growth factors, may be related to membrane ruffling and actin fiber reorganization. PMID- 7499424 TI - Identification of two novel Dictyostelium discoideum cysteine proteinases that carry N-acetylglucosamine-1-P-modification. AB - Dictyostelium discoideum makes multiple developmentally regulated lysosomal cysteine proteinases. One of these, a lysosomal enzyme called proteinase I, contains a cluster of GlcNAc-alpha-1-P-Ser residues. We call this phosphoglycosylation. To study its function, a cDNA library from vegetative cells was screened, and two novel cysteine proteinase clones were characterized (cprD and cprE). Each of them has highly conserved regions expected for cysteine proteinases, but unlike any other, each has a serine-rich domain containing three distinct motifs, poly-S, SGSQ, and SGSG. cprD and cprE cDNAs were overexpressed in Dictyostelium and the active enzymes identified. cprD codes for a protein of approximately 36 kDa (CP4), which is recognized by monoclonal antibodies against GlcNAc-1-P and fucose. cprE corresponds to a 29-kDa protein, which is recognized by antibodies against GlcNAc-1-P. mRNA for both enzymes is present in the vegetative phase and increases during growth on bacteria but decreases throughout development. When the formation of the fruiting body is complete the mRNA for both messages is detected again but in very low levels. Having cloned cDNAs for proteins that carry GlcNAc-1-P should allow us to probe the function of the carbohydrate in these putative lysosomal enzymes. PMID- 7499426 TI - [Cl-]i-dependent phosphorylation of the Na-K-Cl cotransport protein of dog tracheal epithelial cells. AB - Basolateral Na-K-Cl cotransport activity in primary cultures of dog tracheal epithelial cells is stimulated by beta-adrenergic agents, such as isoproterenol, and by apical UTP, which acts through an apical P2-purinergic receptor. While at least part of the stimulatory effect of isoproterenol appears to involve direct activation of the cotransporter via cAMP-dependent protein kinase, cotransport stimulation by apical UTP is entirely secondary to apical Cl- efflux and a resultant decrease in intracellular [Cl-] ([Cl-]i) and/or cell shrinkage (Haas, M., and McBrayer, D. G. (1994) Am. J. Physiol. 266, C1440-C1452). In the secretory epithelia of the shark rectal gland and avian salt gland, Na-K-Cl cotransport activation by both cAMP-dependent and cAMP-independent secretagogues has been shown to be accompanied by phosphorylation of the cotransport protein itself (Lytle, C., and Forbush, B., III (1992) J. Biol. Chem. 267, 25438-25443; Torchia, J., Lytle, C., Pon, D. J., Forbush, B., III, and Sen, A. K. (1992) J. Biol. Chem. 267, 25444-25450). In the present study, we immunoprecipitate the approximately 170-kDa Na-K-Cl cotransport protein of dog tracheal epithelial cells with a monoclonal antibody against the cotransporter of the intestinal cell line T84. Incubation of confluent primary cultures of tracheal cells with isoproterenol and apical UTP increases basolateral-to-apical 36Cl- flux 3.4- and 2.6-fold, respectively, and produces similar increases (3.2- and 2.8-fold, respectively) in 32P incorporation into the approximately 170-kDa cotransport protein. Decreasing [Cl-]i (without concomitant cell shrinkage) by incubating cultures with apical nystatin and reduced apical [Cl-] ([Cl-]alpha) likewise increases both cotransport activity and cotransport protein phosphorylation. These effects become more pronounced with greater reductions in [Cl-]alpha; after 20 min of incubation with nystatin and 32 mM [Cl-]alpha, cotransport activity and 32P incorporation into the cotransport protein are increased 2.8- and 2.7-fold, respectively, similar to increases seen with apical UTP. 2-3-fold increases in cotransporter activity and phosphorylation are also seen in nystatin-treated cells under hypertonic conditions (50 mM sucrose added apically and basolaterally). These findings suggest a close correlation between Na-K-Cl cotransport activity and phosphorylation of the approximately 170-kDa cotransport protein. The latter is phosphorylated in response to both reduced [Cl-]i and cell shrinkage, either or both of which are likely to be involved in secondary cotransport activation in response to apical UTP. PMID- 7499425 TI - The cysteine-rich protein family of highly related LIM domain proteins. AB - Here we describe a family of closely related LIM domain proteins in avian cells. The LIM motif defines a zinc-binding domain that is found in a variety of transcriptional regulators, proto-oncogene products, and proteins associated with sites of cell-substratum contact. One type of LIM-domain protein, called the cysteine-rich protein (CRP), is characterized by the presence of two LIM domains linked to short glycine-rich repeats and a potential nuclear localization signal. We have identified and characterized two evolutionarily conserved members of the CRP family, CRP1 and CRP2, in chicken and quail. Expression of the genes encoding both CRP1 and CRP2 is differentially regulated in normal versus transformed cells, raising the possibility that members of the CRP family may function in control of cell growth and differentiation. PMID- 7499427 TI - Cell surface localization of proteolysis of human endothelial angiotensin I converting enzyme. Effect of the amino-terminal domain in the solubilization process. AB - Angiotensin-converting enzyme (ACE) belongs to the type I class of ectoproteins and is solubilized by Chinese hamster ovary cells transfected with the full length human ACE cDNA. ACE release in Chinese hamster ovary cells involves a proteolytic cleavage occurring in the carboxyl-terminal region, between Arg-1137 and Leu-1138. The subcellular localization of ACE proteolysis was established by pulse-chase experiments, cell surface immunolabeling, and biotinylation of radiolabeled mature proteins. The proteolysis of ACE takes place primarily at the plasma membrane. The solubilization of ACE is less than 2% within 1 h, is increased 2.4-fold by phorbol esters, but is not influenced by ionophores. An ACE mutant lacking the transmembrane domain and the cytosolic part (ACE delta COOH), is secreted at a faster rate without a carboxyl-terminal cleavage, and phorbol esters or ionophores have no effect on its rate of production in the medium. Therefore, the proteolysis of ACE is dependent on the presence of the membrane anchor and suggests that the secretase(s) involved is also membrane-associated. An ACE mutant lacking the amino-terminal domain (ACECF) is secreted 10-fold faster compared with wild-type ACE. The solubilization of ACECF occurs at the plasma membrane and is stimulated 2.7-fold by phorbol esters, and the cleavage site is localized between Arg-1227 and Val-1228. The amino-terminal domain of ACE slows down the proteolysis and seems to act as a "conformational inhibitor" of the proteolytic process, possibly via interactions with the "stalk" of ACE and the secretase(s) itself. PMID- 7499428 TI - The fragile X syndrome single strand d(CGG)n nucleotide repeats readily fold back to form unimolecular hairpin structures. AB - Expansion of a d(CGG)n run within the 5'-untranslated region of the X-linked human gene FMR1 blocks FMR1 transcription, delays its replication, and precipitates the fragile X syndrome. We showed previously that d(CGG)n tracts aggregate into interstrand tetrahelical complexes (Fry, M., and Loeb, L. A. (1994) Proc. Natl. Acad. Sci. U. S. A. 91, 4950-4954). Here we show that these sequences also form under physiological conditions in in vitro unimolecular hairpin structures. Folding is demonstrated by temperature-dependent mobility of d(CGG)n oligomers in a nondenaturing polyacrylamide gel, by UV-hyperchromicity of thermally denaturing oligomers, and by UV cross-linking of compact forms of d(CGG)n runs interspersed by thymidine clusters. That the compact d(CGG)n structures are unimolecular is suggested by their zero-order kinetics of formation. Diethyl pyrocarbonate modification reveals a single, 4-5 residue-long central or epicentral unpaired loop in folded d(CGG)n oligomers. The position of this loop remains unchanged by insertion of thymidine clusters into 15- or 33-mer d(CGG) tracts as indicated by KMnO4 probing of unpaired thymidines. The presence of a single loop in folded d(CGG)n oligomers and the accessibility of every guanine to dimethyl sulfate modification suggest that they are hairpin and not tetraplex structures. Modeling indicates that different d(CGG)n hairpins are stabilized by guanine-guanine Hoogsteen hydrogen bonds or by Hoogsteen and Watson Crick bonds. If formed in vivo, d(CGG)n hairpins could entail slippage and trinucleotide expansion during replication and could obstruct FMR1 gene transcription and replication. PMID- 7499429 TI - In vivo binding of trimethylpsoralen detects DNA structural alterations associated with transcribing regions in the human beta-globin cluster. AB - In order to increase our knowledge about the mechanisms that regulate expression of human beta-like globin genes, we have used a novel technique to analyze the chromatin structure in living cells. This approach allowed us to detect specific DNA regions in vivo where nucleosome folding or unconstrained DNA supercoiling in erythroid cells differs from that in non-erythroid cells. In this method, we use 4,5',8-trimethylpsoralen (TMP) as a probe capable of detecting altered chromatin conformations. Our results show that TMP binds to DNA with a higher affinity over the regions in the locus that are actively expressed, including both the promoter and the transcribed region. This higher affinity detected when comparing erythroid cells with non-erythroid cells does not extend to other regions inside the beta-globin cluster. Our data suggest that the observed effect is likely due to nucleosome displacement. Alternatively, it could result from localized DNA supercoiling, but not from widespread torsional stress across the entire beta like globin locus as hypothesized previously. PMID- 7499430 TI - Cloning of a novel family of mammalian GTP-binding proteins (RagA, RagBs, RagB1) with remote similarity to the Ras-related GTPases. AB - cDNA clones of two novel Ras-related GTP-binding proteins (RagA and RagB) were isolated from rat and human cDNA libraries. Their deduced amino acid sequences comprise four of the six known conserved GTP-binding motifs (PM1, -2, -3, G1), the remaining two (G2, G3) being strikingly different from those of the Ras family, and an unusually large C-terminal domain (100 amino acids) presumably unrelated to GTP binding. RagA and RagB differ by seven conservative amino acid substitutions (98% identity), and by 33 additional residues at the N terminus of RagB. In addition, two isoforms of RagB (RagBs and RagB1) were found that differed only by an insertion of 28 codons between the GTP-binding motifs PM2 and PM3, apparently generated by alternative mRNA splicing. Polymerase chain reaction amplification with specific primers indicated that both long and short form of RagB transcripts were present in adrenal gland, thymus, spleen, and kidney, whereas in brain, only the long form RagB1 was detected. A long splicing variant of RagA was not detected. Recombinant glutathione S-transferase (GST) fusion proteins of RagA and RagBs bound large amounts of radiolabeled GTP gamma S in a specific and saturable manner. In contrast, GTP gamma S binding of GST-RagB1 hardly exceeded that of recombinant GST. GTP gamma S bound to recombinant RagA, and RagBs was rapidly exchangeable for GTP, whereas no intrinsic GTPase activity was detected. A multiple sequence alignment indicated that RagA and RagB cannot be assigned to any of the known subfamilies of Ras-related GTPases but exhibit a 52% identity with a yeast protein (Gtr1) presumably involved in phosphate transport and/or cell growth. It is suggested that RagA and RagB are the mammalian homologues of Gtr1 and that they represent a novel subfamily of Ras homologous GTP binding proteins. PMID- 7499431 TI - Diversity of the pyruvate dehydrogenase kinase gene family in humans. AB - Recent evidence from this laboratory indicates that at least two isoenzymic forms of pyruvate dehydrogenase kinase (PDK1 and PDK2) may be involved in the regulation of enzymatic activity of mammalian pyruvate dehydrogenase complex by phosphorylation (Popov, K.M., Kedishvili, N.Y., Zhao, Y., Gudi, R., and Harris, R.A. (1994) J. Biol. Chem. 269, 29720-29724). The present study was undertaken to further explore the diversity of the pyruvate dehydrogenase kinase gene family. Here we report the deduced amino acid sequences of three isoenzymic forms of PDK found in humans. In terms of their primary structures, two isoenzymes identified in humans correspond to rat PDK1 and PDK2, whereas a third gene (PDK3) encodes for a new isoenzyme that shares 68% and 67% of amino acid identities with PDK1 and PDK2, respectively. PDK3 cDNA expressed in Eschierichia coli directs the synthesis of a polypeptide with a molecular mass of approximately 45,000 Da that possesses catalytic activity toward kinase-depleted pyruvate dehydrogenase. PDK3 appears to have the highest specific activity among the three isoenzymes tested as recombinant proteins. Tissue distribution of all three isoenzymes of human PDK was characterized by Northern blot analysis. The highest amount of PDK2 mRNA was found in heart and skeletal muscle, the lowest amount in placenta and lung. Brain, kidney, pancreas, and liver expressed an intermediate amount of PDK2 (brain > kidney = pancreas > liver). The tissue distribution of PDK1 mRNA differs markedly from PDK2. The message for PDK1 was expressed predominantly in heart with only modest levels of expression in other tissues (skeletal muscle > liver > pancreas > brain > placenta = lung > kidney). In contrast to PDk1 and PDK2, which are expressed in all tissues tested, the message for PDK3 was found almost exclusively in heart and skeletal muscle, indicating that PDK3 may serve specialized functions characteristic of muscle tissues. In all tissues tested thus far, the level of expression of PDK2 mRNA was essentially higher than that of PDK1 and PDK3, consistent with the idea that PDK2 is a major isoenzyme responsible for regulation of pyruvate dehydrogenase in human tissues. PMID- 7499432 TI - A regulatory mechanism that detects premature nonsense codons in T-cell receptor transcripts in vivo is reversed by protein synthesis inhibitors in vitro. AB - Gene rearrangement during the ontogeny of T- and B-cells generates an enormous repertoire of T-cell receptor (TCR) and immunoglobulin (Ig) genes. Because of the error-prone nature of this rearrangement process, two-thirds of rearranged TCR and Ig genes are expected to be out-of-frame and thus contain premature terminations codons (ptcs). We performed sequence analysis of reverse transcriptase-polymerase chain reaction products from fetal and adult thymus and found that newly transcribed TCR-beta pre-mRNAs (intron-bearing) are frequently derived from ptc-bearing genes but such transcripts rarely accumulate as mature (fully spliced) TCR-beta transcripts. Transfection studies in the SL12.4 T-cell line showed that the presence of a ptc in any of several TCR-beta exons triggered a decrease in mRNA levels. Ptc-bearing TCR-beta transcripts were selectively depressed in levels in a cell clone that contained both an in-frame and an out-of frame gene, thus demonstrating the allelic specificity of this down-regulatory response. Protein synthesis inhibitors with different mechanism of action (anisomysin, cycloheximide, emetine, pactamycin, puromycin, and polio virus) all reversed the down-regulatory response. Ptc-bearing transcripts were induced within 0.5 h after cycloheximide treatment. The reversal by protein synthesis inhibitors was not restricted to lymphoid cells, as shown with TCR-beta and beta globin constructs transfected in HeLa cells. Collectively, the data suggest that the ptc-mediated mRNA decay pathway requires an unstable protein, a ribosome, or a ribosome-like entity. Protein synthesis inhibitors may be useful tools toward elucidating the molecular mechanism of ptc-mediated mRNA decay, an enigmatic response that can occur in the nuclear fraction of mammalian cells. PMID- 7499433 TI - Desensitization and internalization of the m2 muscarinic acetylcholine receptor are directed by independent mechanisms. AB - The phenomenon of acute desensitization of G-protein-coupled receptors has been associated with several events, including receptor phosphorylation, loss of high affinity agonist binding, receptor:G-protein uncoupling, and receptor internalization. However, the biochemical events underlying these processes are not fully understood, and their contributions to the loss of signaling remain correlative. In addition, the nature of the kinases and the receptor domains which are involved in modulation of activity have only begun to be investigated. In order to directly measure the role of G-protein-coupled receptor kinases (GRKs) in the desensitization of the m2 muscarinic acetylcholine receptor (m2 mAChR), a dominant-negative allele of GRK2 was used to inhibit receptor phosphorylation by endogenous GRK activity in a human embryonic kidney cell line. The dominant-negative GRK2K220R reduced agonist-dependent phosphorylation of the m2 mAChR by approximately 50% and prevented acute desensitization of the receptor as measured by the ability of the m2 mAChR to attenuate adenylyl cyclase activity. In contrast, the agonist-induced internalization of the m2 mAChR was unaffected by the GRK2K220R construct. Further evidence linking receptor phosphorylation to acute receptor desensitization was obtained when two deletions of the third intracellular loop were made which created m2 mAChRs that did not become phosphorylated in an agonist-dependent manner and did not desensitize. However, the mutant mAChRs retained the ability to internalize. These data provide the first direct evidence that GRK-mediated receptor phosphorylation is necessary for m2 mAChR desensitization; the likely sites of in vivo phosphorylation are in the central portion of the third intracellular loop (amino acids 282-323). These results also indicate that internalization of the m2 receptor is not a key event in desensitization and is mediated by mechanisms distinct from GRK phosphorylation of the receptor. PMID- 7499434 TI - Binding of tenascin-C to soluble fibronectin and matrix fibrils. AB - The small splice variant of tenascin-C (TN) has eight fibronectin type III (FN3) domains. The major large splice variant has three (in chicken) or seven (in human) additional FN3 domains inserted between domains five and six. Chiquet Ehrismann et al. (Chiquet-Ehrismann, R., Matsuoka, Y., Hofer, U., Spring, J., Bernasconi, C., and Chiquet, M. (1991, Cell Regul. 2, 927-938) demonstrated that the small variant bound preferentially to fibronectin in enzyme-linked immunosorbent assay, and only the small variant was incorporated into the matrix by cultures of chicken fibroblasts. Here we have studied human TN, and confirmed that the small variant binds preferentially to purified fibronectin and to fibronectin-containing extracellular matrix. Thus this differential binding appears to be conserved across vertebrate species. Using bacterial expression proteins, we mapped the major binding site to the third FN3 domain of TN. Consistent with this mapping, a monoclonal antibody against an epitope in this domain did not stain TN segments bound to cell culture matrix fibrils. The enhanced binding of the small TN variant suggests the existence of another, weak binding site probably in FN3 domains 6-8, which is only positioned to bind fibronectin in the small splice variant. This binding of domains 6-8 may involve a third molecule present in matrix fibrils, as the enhanced binding of small TN was much more prominent to matrix fibrils than to purified fibronectin. PMID- 7499435 TI - Conservation of ligand specificity between the mammalian and amphibian fibroblast growth factor receptors. AB - We have previously cloned and sequenced a newt keratinocyte growth factor receptor (KGFR) cDNA which exhibited a unique spatial and temporal expression pattern in the regenerating newt limb. In this report, we further characterize the biochemical and functional properties of this newt KGFR. A stable Chinese hamster ovary transfectant overexpressing the newt KGFR was capable of binding both 125I-fibroblast growth factor-1 (FGF-1) and 125I-FGF-7 but not 125I-FGF-2, indistinguishable from the human KGFR. Scatchard analysis and cross-linking studies further support the conclusion that FGF-1 and FGF-7 are the ligands for the newt KGFR. In addition to their ability to bind to FGFs, both the human and the newt KGFR are also capable of repressing differentiation in mouse MM14 myoblasts. MM14 cells express FGFR1 and are repressed from differentiation by FGF 1, FGF-2, and FGF-4 but not FGF-7. Co-transfection of MM14 cells with either a human or newt KGFR expression construct conferred a response to FGF-7 as determined by a human alpha-cardiac actin/luciferase reporter construct. The response to FGF-7 was similar to the endogenous FGF response as FGF-7 prevented MM14 myoblasts from undergoing terminal differentiation. Thus, both the human and the newt KGFRs transduce signals similar to those transduced via the endogenous mouse FGFR1. Together these data indicate that this newly isolated newt KGFR is a functional receptor as it binds two FGF family members with high affinity and mediates signaling in skeletal muscle myoblasts. Because the binding pattern of the newt KGFR is similar to the pattern observed for its mammalian counterpart, it emphasizes the strict conservation that this ligand/receptor system has undergone through evolution. PMID- 7499436 TI - Unique expression of major histocompatibility complex class I proteins in the absence of glucose trimming and calnexin association. AB - Recent evidence indicates that efficient expression of major histocompatibility complex (MHC) complexes requires their interaction with the resident endoplasmic reticulum (ER) chaperone calnexin, which for certain proteins functions as a lectin specific for monoglucosylated glycans. In the current report, we studied the expression of MHC class I proteins in BW wild type thymoma cells (BW WT) and glucosidase II-deficient BW PHAR2.7 cells. Consistent with a requirement for glucose (Glc) trimming for interaction of class I proteins with calnexin, we found that nascent H-2Kk proteins associated with calnexin in untreated BW WT cells, but not in BW WT cells treated with the glucosidase inhibitor castanospermine (cas), or in untreated glucosidase II-deficient BW PHAR2.7 cells. Surprisingly, we found that H-2Kk expression occurred with similar efficiency in BW PHAR2.7 cells as in BW WT cells and that formation of nascent H-2Kk complexes was perturbed by cas treatment in BW WT cells but not in BW PHAR2.7 cells. Finally, it was noted that expression of the molecular chaperone Bip was markedly increased in BW PHAR2.7 cells relative to BW WT cells, which is suggested to play a role in regulating the expression of H-2Kk complexes in BW PHAR2.7 cells. The current study demonstrates that Glc trimming is required for efficient interaction of nascent H-2Kk proteins with calnexin; that expression of MHC class I proteins can, under certain conditions, proceed effectively in the absence of Glc trimming and calnexin association; and that Bip expression is markedly increased under conditions where diglucosylated glycans persist on nascent glycoproteins within the ER. These data are consistent with the hypothesis that alternative oligomerization pathways exist for class I proteins within the quality control system of the ER that have differential requirements for removal of Glc residues from nascent glycan chains. PMID- 7499437 TI - Amino acid substitutions in the herpes simplex virus transactivator VP16 uncouple direct protein-protein interaction and DNA binding from complex assembly and transactivation. AB - The herpes simplex virus transactivator VP16 directs the assembly of a multicomponent protein-DNA complex that requires the participation of two cellular factors, the POU homeodomain protein Oct-1, which binds independently to response elements, and VCAF-1 (VP16 complex assembly factor; also called HCF, C1), a factor that binds directly to VP16. A number of distinct properties of VP16 have been implicated in the assembly of the VP16-induced complex (VIC). These include its independent association with VCAF-1 and, under appropriate conditions, its ability to bind to DNA or to DNA-bound Oct-1 in the absence of VCAF-1. In order to probe the requirements of these individual interactions in the functional assembly of VIC, we mutated selected charged amino acids in two subdomains of VP16 previously shown to be important in protein-DNA complex formation. Purified VP16 proteins were analyzed for their ability to direct protein-DNA complex formation and to interact directly with VCAF-1. Several classes of mutants that were differentially compromised in VCAF-1 interaction, direct DNA binding, and/or association with DNA-bound Oct-1 were obtained. Interestingly, all of the derivatives were still capable of generating the VIC complex in vitro and activating transcription in vivo. Our findings indicate that the cooperative assembly of functional VP16-containing complexes can occur by pathways that do not necessarily require the prior interaction of VP16 with VCAF 1 or the ability of VP16 to bind directly to DNA or associate with DNA-bound Oct 1. PMID- 7499438 TI - Cloning and functional expression of a cDNA encoding a human type 2 neuropeptide Y receptor. PMID- 7499439 TI - Cancer genetic risk assessment: a note of caution. PMID- 7499442 TI - In vitro investigation of a combination of two drugs, cisplatin and carboplatin, as a function of the area under the c/t curve. AB - The log/log relationship between the IC50 of cisplatin or carboplatin and the exposure time, determined by human tumor clonogenic assay (HTCA) and MTTAI (3 (4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay with additional incubation) using PC-14 cells, exhibited a straight line with a slope of -1, indicating that both drugs have AUC-dependent cytotoxicity (AUC, area under the c/t curve). The combined effect of cisplatin and carboplatin was estimated by the median-effect analysis using HTCA, and it was additive when the AUC ratio (AUC of free platinum from carboplatin/that from cisplatin) was low (3.2, 6.5 or 13.1 in each of PC-7, PC-9, PC-14, H-69, and K562). However, it was significantly worse at a higher AUC ratio (19.5 in PC-7, PC-9, and PC-14). The log/log relationship of each drug, determined by MTTAI using human bone marrow cells, showed that each drug exerts an AUC-dependent cytotoxicity on marrow granulocytes. When cisplatin and carboplatin were combined at an AUC ratio of 14, the therapeutic index was significantly better than that of carboplatin alone and less than that of cisplatin alone using K562, PC-9 and PC-14, indicating the usefulness of this combined therapy for tumor cells with high sensitivity to platinum compounds at this AUC ratio. PMID- 7499441 TI - Inhibitory effect of hydroxyflutamide plus tamoxifen on oestradiol-induced growth of MCF-7 breast cancer cells. AB - The antiandrogen flutamide has been reported to exert antiproliferative actions on breast cancer both in vitro and in vivo. Here we study the action of its active metabolite hydroxyflutamide on the oestradiol-induced growth of MCF-7 breast cancer cells. The results show that the antiandrogen inhibits the cell growth. Moreover hydroxyflutamide adds its antiproliferative effect to the action of the antioestrogen tamoxifen. The inhibitory effect is dose-dependent and it is unaffected by tamoxifen concentrations up to levels able to block oestrogen receptors completely. Dihydrotestosterone experiments parallel those on hydroxyflutamide. When the two substances are administered together, neither antagonistic nor additive effects are appreciable. Data are consistent with an androgen-like action of hydroxyflutamide on breast cancer cells. The antiproliferative effect of hydroxyflutamide, without virilizing side-effects, suggests that it is worth exploring its possible employment together with antioestrogens in the treatment of breast cancer patients. PMID- 7499443 TI - Induction of antitumor immunity and treatment of preestablished tumor by interleukin-6-gene-transfected melanoma cells combined with low-dose interleukin 2. AB - Transfer of cytokine genes into tumor cells has proven a valuable approach for cancer treatment. In order to generate a more effective cancer vaccine, we transfected the human interleukin-6 (IL-6) gene into B16 melanoma cells. A B16 cell clone secreting the highest level of IL-6 was obtained by G418-resistant selection, limiting dilution and IL-6 assay. The IL-6-gene-transfected tumor cells exhibited in vitro growth inhibition, reduced tumorigenicity and decreased metastatic competence. After immunization with the inactivated IL-6-gene transfected vaccine, the murine cytotoxic T lymphocyte activity, natural killer activity and lymphokine-activated killer activity increased markedly. After treatment with the vaccine, the tumor-bearing mice showed significant growth inhibition of subcutaneous tumor, reduction in pulmonary metastases and extension of survival time. The above therapeutic effect was better when low-dose IL-2 was administered simultaneously, although this dosage of IL-2 had no in vivo antitumor effect. These data demonstrated that IL-6-gene-transfected cancer vaccine has a potent antitumor effect via efficient induction of antitumor immunity, and a better therapeutic effect could be achieved when the vaccine is combined with low-dose IL-2 as adjuvant. PMID- 7499440 TI - Cytokines and bone marrow transplantation. AB - This article summarizes the various current uses of cytokines, and bone marrow and peripheral blood stem cell transplantation for cancer treatment. Interferons, interleukin-2 and hematopoietic growth factors are extensively used for the treatment of hematopoietic as well as nonhematopoietic malignancies, either as the main therapeutic agents or as an adjuvant. Research on gene therapy using cytokines is in progress. Stem cell transplantation is also discussed in the context of gene therapy of cancer. Gene therapy is expected to become a future modality in cancer treatment. PMID- 7499444 TI - Induction of apoptosis in cultured retinoblastoma cells by the protein phosphatase inhibitor, okadaic acid. AB - The induction of apoptosis in cultured retinoblastoma cells by diverse drugs was examined by analyzing DNA fragmentation, a hallmark of apoptosis. First, the ability of six retinoblastoma cell lines to undergo apoptosis was surveyed using etoposide (30 micrograms/ml, 20 h exposure). The NCC-RbC-60 cell line, established in this laboratory showed DNA fragmentation clearly, whereas the other cell lines tested, including the representative retinoblastoma cell line, Y 79, did not show distinct DNA fragmentation. Biochemical modulators, such as A23187, forskolin, retinoic acid, phorbol 12-myristate 13-acetate and okadaic acid, were examined to ascertain whether they could induce apoptosis in NCC-RbC 60 and Y-79 cells after exposure for 20 h. Only okadaic acid induced DNA fragmentation in all the retinoblastoma cell lines tested and it induced DNA fragmentation in Y-79 cells in a time- and concentration-dependent manner. Flow cytometric analysis and microscopic examination revealed that Y-79 cells treated with okadaic acid for 24-48 h accumulated at the G2/M, especially M, phases, before undergoing DNA fragmentation. Other mitotic poisons, nocodazole, colcemid and taxol, also induced apoptosis in Y-79 cells. In the K1034 cell line, established from non-malignant retinal pigmented epithelium, okadaic acid failed to induce both G2/M arrest and DNA fragmentation. These findings suggest that okadaic-acid-induced apoptosis occurs as a result of metaphase arrest. PMID- 7499445 TI - Fructose 2,6-bisphosphate metabolism in Ehrlich ascites tumour cells. AB - Cancer cell energy metabolism is characterized by a high glycolytic rate, which is maintained under aerobic conditions. In Ehrlich ascites tumour cells, the concentration of fructose 2,6-bisphosphate (Fru-2,6-P2), the powerful activator of 6-phosphofructo-1-kinase, is tenfold increased. The bifunctional enzyme 6 phosphofructo-2-kinase/fructose-2,6-bisphosphatase (PFK-2/FBPase-2), synthesizing and degrading Fru-2,6-P2, was characterized. The molecular mass is 120 kDa. The dependence of PFK-2 activity on the substrate concentrations is hyperbolic (Km for Fru-6-P = 0.09 mM; Km for ATP = 0.7 mM), while the dependence of the FBPase-2 activity on the concentrations of Fru-2,6-P2 is sigmoidal (K0.5 for Fru-2,6-P2 = 4 microM). The PFK-2/FBPase-2 activity ratio is 1. PFK-2 activity is inhibited by citrate (I0.5 = 0.17 mM) and phosphoenolpyruvate (I0.5 = 0.08 mM) but only weakly by glycerol 3-phosphate (I0.5 = 1.57 mM). In contrast to the liver enzyme, the activity of tumour PFK-2/FBPase-2 is not influenced by the action of cAMP dependent protein kinase. The kinetic properties as well as ion-exchange chromatography pattern differ from their normal counterparts in liver and muscle. The properties are likely to contribute to the maintenance of the high glycolytic rate in these tumour cells. PMID- 7499447 TI - Prognostic factors for survival in patients with advanced renal cell carcinoma treated with interleukin-2 and interferon-alpha. AB - A group of 73 patients with advanced renal cell carcinoma, treated in different phase II trials with interferon alpha and/or interleukin-2, have been evaluated to identify potential baseline prognostic factors predicting their survival. The eligibility criteria were very similar across studies and included ECOG performance status < or = 2, measurable or evaluable disease and no CNS metastases. The overall response rate was 8%. The overall survival was 33% at 2 years and 18% at 1 year. In the univariate analysis three prognostic factors were correlated with disease outcome: ECOG performance status (0 versus > or = 1), time from diagnosis to treatment (< or = 12 months versus > 12 months) and number of metastatic sites (1 versus > or = 2). Multivariate analysis identified ECOG performance status and number of metastatic sites as important prognostic factors for survival. The true impact on patient survival of the selection of patients rather than the treatment itself should be evaluated in controlled trials. PMID- 7499448 TI - International conference on cell interactions in malignancy, development and differentiation. Heidelberg, Germany, April 1995. PMID- 7499446 TI - Enhanced cellular proliferative activity and cell death in chronic thyroiditis and thyroid papillary carcinoma. AB - For the analysis of cellular proliferative activity and cell death in thyroid diseases, the Ki-67 labeling index, bcl-2 protein expression and cell death of follicular epithelia by immunohistochemistry and in situ DNA nick-end labeling methods were evaluated in normal thyroid tissues as well as in surgical specimens from cases of Hashimoto's disease (16 cases), focal lymphocytic thyroiditis (13 cases), Graves' disease (15 cases), follicular adenoma (20 cases) and papillary carcinoma (43 cases). Cellular proliferative activity and cell death were both enhanced in cases of thyroiditis, including Hashimoto's disease and focal lymphocytic thyroiditis. Thyroids from patients with follicular adenoma and papillary carcinoma also showed increased cellular proliferative activity and cell death. In addition, predominant high cellularity and partial loss of bcl-2 protein expression in papillary carcinoma suggested that the overgrowth and dedifferentiation were associated with malignancy. PMID- 7499449 TI - [Treatment of hepatocellular carcinoma in the presence of liver cirrhosis]. AB - Hepatocellular carcinoma accounts for 90% of the primary malignant liver tumours. Most cases occur in cirrhotic livers. Management decisions should not be based on the stage of the tumour extension but rather on the functional situation of the liver. The first therapeutic option which should be considered is surgical resection if the case presents with a single tumour (or less than 4 tumours for some teams) without detectable metastasis nor intraportal thrombosis and if the liver remaining after surgery will be sufficient for normal hepatic functions. The disadvantage of resection is the high risk of recurrence in the long term. Liver transplantation cannot be proposed if the hepatocellular carcinoma has produced clinical signs but it can be a possibility in case of a resectable tumour in the framework of a prospective protocol comparing transplantation and resection. Intra-arterial injection of 131-iodine linked lipiodol is the only effective treatment in case of portal thrombosis. Chemoembolization of nonresectable hepatocellular carcinoma has led to spectacular tumour response but its effect on survival has not been demonstrated by randomized studies. For tumours less than 3 cm in diameter, even multifocal alcoholization has proved encouraging results. Although a randomized study of questionable quality suggested tamoxifen could be effective, there is no current indication for this drug. External radiotherapy may be a possibility in the future, especially with proton irradiation. Thus the current management of hepatocellular carcinoma is in a difficult, even paradoxical, situation since there is a wide therapeutic choice (resection, alcoholization, transplantation) for the rare cases with small tumours but almost no possibilities for the more severe cases most frequently encountered. PMID- 7499450 TI - [Cervical paragangliomas. Results apropos of a series of 33 patients]. AB - During the period 1971-1993, 33 patients were treated for cervical chemodectomas, 41 tumors were diagnosed. Seven patients had multicentric tumors. All patients expect one with bilateral carotid tumor were treated surgically. A carotid resection was performed in 4 patients. Six patients were given radiation therapy: five patient after surgery and one treated by irradiation alone. During the follow-up period one patient died of bone metastasis 6 year after a partial surgery and radiation therapy, and one had local recurrence treated with irradiation. The surgical management is the major treatment of these tumors, it revealed no postoperative cerebrovascular accident and limited complications secondary to unavoidable nerves sacrifice. Radiation therapy is performed only on particular cases: surgery contra-indications and nonradical resection. PMID- 7499451 TI - [Splenectomy for hematologic disease in patients over 70 years of age. Apropos of 38 cases]. AB - A retrospective series of 38 splenectomies performed in patients over 70 for haematology diseases is reported. There were no post-operative deaths and 8 patients (21%) had complications which resolved favourably. Paradoxically, mortality and morbidity were lower in elderly patients although the difference was not statistically significant. No factor predictive of post-operative outcome was found. It was concluded that age is not a contraindication although the clinical situation is a predominant factor in patient selection. PMID- 7499452 TI - [Acute solitary diverticulitis of the cecum, a rare cause of acute abdomen. Apropos of 9 cases and review of the literature]. AB - The natural history of colonic diverticulitis rarely includes acute solitary localization in the cecum. Complications are even more rarely seen. We report retrospectively nine cases seen in our unit. There were 5 males and 4 females, mean age 54 years. The patients presented with a more or less painful abdomen with localization in the right iliac region and signs of peritoneal irritation. Physical examination revealed a painful mass in this zone. Fever and hyperleukocytosis were not found in all patients. Barium enema demonstrated signs of pseudocompression of the cecum in four cases suggesting a tumoral formation in two and diverticulitis in the two others. A right hemicolectomy was performed in five patients as peroperative pathological diagnosis could not be ascertained on the macroscopic specimen. Diverticulectomy in the four other patients was unsuccessful in direct relation with the disease course and late diagnosis. PMID- 7499453 TI - [Post-traumatic liponecrosis. Apropos of a clinical case]. AB - Localized post-trauma liponecrosis, also called lipoatrophy, is a benign disease with no functional consequences but does cause important aesthetic problems. Diagnosis is often missed. Necrosis of the adipocytes occurs after direct trauma and absorption of the haematoma. It is important to search for favouring factors and order the necessary examinations. The therapeutic strategy is presented. PMID- 7499454 TI - [Voluminous retroperitoneal liposarcoma. Apropos of a case and review of the literature]. AB - The authors report a case of bulky retroperitoneal liposarcoma (6 kg) in a 42 year-old woman. This observation of liposarcoma is interesting in that diagnosis was incidental during check up for recent hypertension. The diagnosis was suggested by ultrasonography, essentially computed tomography and suspected by multiple subcutaneous lipomas. Treatment was exclusively surgical with complete tumor resection including the left adrenal gland. The authors review literature data of retroperitoneal liposarcoma. Because of frequent recurrency, follow-up of tumor resection must be made by CT scanning every six months. An aggressive surgical approach including adjacent organs if necessary and total excision of local recurrencies can improve long-term prognosis. Postoperative adjuvant radio- and/or chemotherapy does not effect survival. PMID- 7499455 TI - [Internal hernia of the small intestine through a breach of the broad ligament, apropos of a case. Review of the literature]. AB - Strangulated herniation of bowel occurring through a defect of the broad ligament is extremely rare. A recent episode of such a herniation of small bowel through the left mesosalpynx is presented. A review of literature reveals 61 cases previously reported. A classification with 4 types of defects is described. Without antecedent of uterine surgery, delivery trauma and congenital abnormalities are considered to be causes of these defects although real mechanism is unknown. Preoperative CT scan is helpful in suggesting the diagnosis: pelvic loop dilatation, uterine lateral deviation. Surgical management includes reperitonealization of the defect and inspection of the controlateral broad ligament. PMID- 7499456 TI - [A rare cause of massive upper digestive hemorrhage. Duodenal tuberculosis, apropos of two cases]. AB - Tuberculosis of the duodenum was revealed in two patients by severe upper digestive tract bleeding. These cases illustrate the difficulties in diagnosing tuberculosis of the duodenum which can present as a simple bleeding ulceration. This "infectious" ulcer is an exceptional finding but certain diagnosis can be made at laparotomy with extemporaneous pathology of the lymph nodes. For non complicated cases, the diagnosis may be suspected on the bases of routine laboratory tests. Surgery should be limited as much as possible since specific antibiotics are usually successful in cases of digestive tract tuberculosis. PMID- 7499458 TI - [Epidural anesthesia and postoperative analgesia]. PMID- 7499457 TI - [Melanosarcoma of the head of the pancreas with primary appearance]. PMID- 7499459 TI - [Motoneuronal death following prevention of reinnervation during postnatal development]. AB - The hypothesis that survival of motoneurons depends on targets postnatally was tested in Wistar rats of known age. Under ether anesthesia, the cut distal end of a medial gastrocnemius (MG) nerve was anchored to the lateral gastrocnemius (LG) muscle to prevent reinnervation. The compound action potential (CAP) in response to stimulation of the MG nerve was recorded from L4 and L5 ventral roots to evaluate the degree of motoneuronal death in the nerve. CAPs of MG nerves were greatly reduced 1 to 8 months after the operation in the rats operated on at 6 or 9 days of age, which indicates that the majority of MG motoneurons died after the operation. In contrast, the CAPs in the animals which were operated on at older than 12 days were similar to normal animals. However, the number of motor units that remained in the implanted MG nerve was 1.3 on the average in the rats operated on at 12-days-old. In normal rats, it was 87. Thus, the results indicate that in the rats operated on at 12 days of age, a majority of MG motoneurons survived without making functional synapses. We conclude that there is a sharp critical period, between 9 and 12 days after birth, during which motoneuronal target dependency changes. PMID- 7499460 TI - [Difference in MRI findings and risk factors between multiple infarction without dementia and multi-infarct dementia]. AB - MRI findings and risk factors for vascular dementia were evaluated with multi variate analysis in 96 multi-infarct patients without dementia and 40 multi infarct patients with dementia (MID). Only subjects with small infarcts in the territory of the perforator artery or deep white matter were studied. The diagnosis of MID was diagnosed according to DMS-III criteria and Hachinski's ischemia score. Location and area of patchy high-intensity areas including small infarcts, the degree of periventricular high intensity (PVH), and the degree of brain atrophy were examined with MR images. Independent variables were: history of hypertension, diabetes mellitus, other complications; systolic and diastolic blood pressure, atherosclerotic index, hematocrit, history of smoking, level of education, and activities of daily life (ADL). Hayashi's quantification method II was used to analyze the data. The most significant correlation was found between history of hypertension and dementia (partial correlation coefficient: 0.39). Significant correlations were also found between ADL and dementia (0.32), between thalamic infarction and dementia (0.31), and between PVH and dementia (0.27). Age, brain atrophy index, and history of diabetes mellitus contributed little to dementia. The contribution to dementia did not differ significantly between right and left patchy high-intensity areas on MR images. Location of infarcts, except for bilateral thalamic infarcts and large PVH, contributed little to dementia. Thus it would be difficult to base a prediction of the prevalence of vascular dementia on MRI findings. However, both hypertension and ADL contribute to vascular dementia and both are treatable, which may be significant for the prevention of dementia. PMID- 7499462 TI - Immunohistochemical investigation of hepatocellular pi class glutathione S transferase in normal and regenerating rat liver. AB - We have immunohistochemically investigated the occurrence of pi class glutathione S-transferase (GST-pi) both in normal rat liver and in rat liver regenerating after hepatectomy. We made our observations by examining frozen liver sections rather than the paraffin sections used in previous studies. First we examined normal liver tissue. Light microscopic examination revealed weak, discontinuous immunofluorescence along the hepatocellular plasma membranes, indicating the presence of GST-pi. Then, using immunoelectron microscopy, we detected immuno reaction products along the bile canalicular plasma membranes, indicating the location of the GST-pi more precisely. On the second and third days after hepatectomy, as the liver was regenerating, we observed that both the immunofluorescence and the reaction products were much more intense and that the bile canaliculi were dilated as well. These observations indicate that in regenerating liver, GST-pi which is a cytosolic enzyme, concentrates in the cytosol of the hepatocytes directly under the bile canalicular plasma membranes. These observations are the first reported morphological confirmation of induced GST-pi in regenerating liver. PMID- 7499461 TI - [Anatomical study of the lumbar multifidus muscle and its innervation in human adults and fetuses]. AB - The lumbar multifidus muscle was dissected with particular reference to its innervation in 10 Japanese adults and 10 fetuses from 5 to 10 months old. The results were as follows: 1) The multifidus muscle attached to the spinous process of a lumbar vertebra was segmentally innervated by the medial branch of the dorsal ramus of the lumbar nerve on a level with the spinous process. 2) The motor point of the nerve innervating the multifidus muscle in adults was slightly ventro-caudal to that portion of the lumbar spinous process with muscle attached. 3) The multifidus muscle in fetuses was thinner and flatter than that in adults. The motor point of the nerve innervating this muscle was slightly caudal at the lateral margin of the muscle to that portion of the lumbar spinous process with the muscle attached. These differences can be attributed to the fact that muscles used to oppose gravity are undeveloped in fetuses. 4) There was variation in 5 fascicles in total, in which the medial branch of the dorsal ramus of a lumbar nerve entered the multifidus muscle originating at the spinous process of a lower lumbar vertebra. PMID- 7499463 TI - [Experimental study and clinical application of a new combination chemotherapy with cis-platinum, adriamycin and carboquone in patients with advanced prostate cancer]. AB - In vitro chemosensitivity of established cell lines from human prostate cancer (8 PC 93, 19 PC 93, DU 145 and PC 3) to various anticancer drugs was examined by clonogenic assay. The drugs used were aclarubicin (ACR), adriamycin (ADM), carboquone (CQ), vincristine (VCR), ifosfamide (IFM), peplomycin (PEP) and cis platinum (CDDP). To compare antitumor activity of different drugs, the predicted anticancer activity (PAA) was calculated from the 50% inhibition doses and the peak plasma concentration of the clinically used dose. High antitumor activity of drug was considered if PAA > or = 1 was observed. The chemosensitivities were: CQ > ADM > CDDP > VCR > PEP > IFM > ACR in the clonogenic assay. 19 PC 93 and DU 145 were sensitive to CQ, ADM and CDDP, but 8 PC 93 and PC 3 were sensitive to CQ, ADM, CDDP and VCR. Thus, a new combination chemotherapy with CDDP, ADM and CQ (PAQ therapy) was used clinically. PAQ therapy was given to sixteen patients with stage D advanced prostate cancer. Of these patients, 14 were undergoing relapse from antiandrogen therapy and 2 had hormone-resistant prostate cancer. The mean interval from the start of the prior treatment to relapse of the cancer was 18 months. The effectiveness of the new therapy was judged according to the response criteria for prostate cancer treatment of Japan. Three patients showed partial response (PR), 9 were stable disease (Stable) and 4 showed progressive disease (PD). The mean response duration in the patients with PR and with Stable was 11.6 months. The survival length of the responders (PR + Stable) was significantly longer than that of the nonresponders (PD) (p < 0.001). The side effect of PAQ therapy was lower than the moderate degree. Therefore, we considered PAQ therapy to be one of the clinical trials for the treatment of advanced prostate cancer. PMID- 7499465 TI - [Morphological study of cardiac veins that drain into the coronary sinus, with special reference to the coronary artery dominant pattern]. AB - The morphology of the cardiac veins that drain into the coronary sinus was studied in 31 human hearts, with special reference to the coronary artery dominant pattern (Barnes' classification). The results were as follows: 1. Great cardiac vein (GCV) The average GCV index of Barnes type I (BI) did not differ significantly from that of Barnes type II (BII). 2. Left marginal vein (LMV) (1) The LMVs were divided into two types, according to the angle made by the LMV and the truncal vein: rect (18 cases, 58%) and acute (11 cases, 38%) angular influx type. (2) The average LMV angle of BI (n = 9, mean +/- SD 77.0 +/- 23.2) was greater than that of BII (n = 19, mean +/- SD 48.1 +/- 34.3) and the difference was statistically significant (p < 0.05). (3) The average LMV index of BI did not differ significantly from that of BII. 3. Left posterior ventricular vein (LPVV) (1) The average LPVV influx angle of BI did not differ significantly from that of BII. (2) The average LPVV index of BI did not differ significantly from that of BII. 4. Middle cardiac vein (MCV) The average MCV index of BI did not differ significantly from that of BII. 5. Small cardiac vein (SCV) The average SCV index of BI did not differ significantly from that of BII. 6. Anterior cardiac veins (ACVs) The number of ACVs correlated with the SCV index, and the correlation was statistically significant (n = 31, r = -0.416, p < 0.02). 7. Coronary sinus (CS) The average CS index of BI did not differ significantly from that of BII. The right ventricular wall may not be perfused effectively during open heart surgery when retrograde coronary sinus perfusion (RCSP) is used. The present study demonstrated that this method can produce poor right ventricular perfusion in cases when the ACVs developed well. PMID- 7499464 TI - Cell damage and liberation of nitric oxide synthase in rat heart induced by endotoxin administration. AB - To evaluate the relationship between cardiovascular injury and the pathological aspect of the liberation of the nitric oxide synthase in endotoxin shock, Wistar rats were injected intraperitoneally with 10 mg/kg E. coli endotoxin and the cardiovascular changes were observed chronologically by immunohistochemical and electron microscopic techniques at 2, 4, 8 and 12 h respectively after endotoxin administration. Light microscopically, irregular contraction of cardiomyocytes was observed in the early stage. After 8 h of endotoxin administration, occasional disintegration of the myocytes as well as lysis of myofibrils became prominent, and diminished stainings of myoglobin and actin were seen in these myocytes. Ultrastructurally, increased pinocytotic vesicles were observed in endothelial cells associated with widened intercellular spaces, and vacuolization of endothelial and medial cells of the vasculature. In cardiomyocytes, swelling of mitochondria and interstitial edema were noted at 12 h after endotoxin administration. Immunohistochemically, an increased permeability of albumin was noted in the myocytes and vasculature and nitric oxide synthase was localized on the cytomembrane of the endothelium of the coronary arteries in control rats. After endotoxin administration, the reaction products of nitric oxide synthase increased in the endothelial and medial cells, and cardiomyocytes. These results suggest that the increased activity of nitric oxide synthase is an important factor in the excessive production of nitric oxide and in promoting the pathologic changes in the cardiovascular system after endotoxin administration. PMID- 7499467 TI - [Current status and future prospects of molecular medicine]. PMID- 7499466 TI - [Analysis of numerical chromosomal aberrations in bladder cancer detected by fluorescence in situ hybridization]. AB - The recently introduced fluorescence in situ hybridization method (FISH method) is a chromosomal analysis for examining interphase nuclei and seems to be a promising approach for chromosomal analysis of a solid tumor, which is considered hard to deal with by a conventional chromosomal staining method. In the present study, a bladder tumor (transitional cell carcinoma) was evaluated by chromosomal analysis, including interphase nuclei, with this FISH method to examine chromosomal numerical aberration in relation to the grade and stage, expecting successful, accurate determination of reliable prognostic factors based on its relapse and invasive intensity. Tissue specimens obtained from 20 patients with bladder tumor (transitional cell carcinoma) were used. The probe used in this study was specific for chromosome No. 1, 7, and 9, which were frequently indicated as being abnormal by a conventional chromosomal staining method. As a result, a tendency towards polysomy was observed in chromosome No. 1 and 7 of high grade invasive cancer cases, while a tendency towards monosomy was evident in chromosome No. 9 of low grade noninvasive cancer cases. From the results mentioned above and consideration of related literatures, it was suggested that monosomy of chromosome No. 9 is important as an index for knowing changes in the initial developmental stage of bladder tumor and polysomy of chromosome No. 1 and 7 is important as an index for knowing changes in the progressive stage. PMID- 7499468 TI - [DNA-based diagnosis of breast cancer cases]. PMID- 7499469 TI - [Genetic diagnosis of mitochondrial diseases]. PMID- 7499470 TI - [Recombinant virus vectors for human gene therapy]. PMID- 7499471 TI - [Gene therapy for cancer]. PMID- 7499472 TI - [Gene therapy for AIDS]. PMID- 7499473 TI - [RAS oncogene in hematological malignancies]. PMID- 7499474 TI - [Series 5. Surgical treatment of Graves' disease. Regarding remnant thyroid amount and postoperative thyroid function]. PMID- 7499475 TI - [Self-defence power fighting against AIDS. Cellular immunity and its activation]. PMID- 7499476 TI - Fluorescence polarization study of the rigor complexes formed at different degrees of saturation of actin filaments with myosin subfragment-1. AB - A serine residue located in the active site of myosin head (S1) was labelled by 9 anthroylnitrile, an amino group located in the central domain of S1 was labelled by 7-diethylamino-3-(4'-isothio-cyanato-phenyl)-4-methylcoumari n, a cysteine residue located near the C-terminus of S1 was labelled by 5-[2-((iodoacetyl) amino)ethyl]-amino-naphthalene-1-sulfonic acid (1,5-IAEDANS) and a cysteine residue located near the C-terminus of the alkali light chain 1 was labelled with iodoacetamido-tetramethyl-rhodamine. Polarization of fluorescence of S1 was measured in solution (where it indicated the mobility of actin-bound S1) and in myofibrils (where it indicated orientation of probes) to check whether the anisotropy of S1 labelled at different positions depended on the molar ratio S1:actin. In solution, when increasing amounts of actin were added to a fixed amount of labelled S1 (i.e. when myosin heads were initially in excess over actin), anisotropy saturated at 1 mol of S1 per 1 mol of actin. When increasing amounts of S1 were added to a fixed amount of F-actin (i.e. when actin was initially in excess over S1), the anisotropy saturated at 1 mol of S1 per 2 mols of actin. In myofibrils, orientation of S1 was different when S1 was added at nanomolar concentration (intrinsic actin was in excess over extrinsic S1) then when it was added at micromolar concentration (excess of S1 over actin). The fact that the anisotropy of S1 labelled at different positions depended on the molar ratio excluded the possibility that changes were confined to one part of the cross-bridge and supports our earlier proposal that the two rigor complexes which S1 can form with F-actin differ globally in conformation. PMID- 7499477 TI - Expression of extraocular myosin heavy chain in rabbit laryngeal muscle. AB - The intrinsic laryngeal muscles of mammals are functionally heterogeneous, some of these muscles (e.g. the thyroarytenoid) contract extremely rapidly, like extraocular muscle, whilst others (e.g. the cricothyroid) contract as fast as limb fast muscle. The extraordinarily rapid contraction speed of extraocular muscles is associated with a fast myosin not found in limb muscles. In this work we explored the possibility that the thyroarytenoid muscle may also express this extraocular-specific fast myosin by raising a monoclonal antibody (mab 4A6) against its heavy chain. Electrophoretic separation of native isomyosins revealed that both the extraocular and the thyroarytenoid have two similar bands migrating ahead of bands found in limb fast or cricothyroid myosins. These two bands bound mab 4A6. The thyroarytenoid muscle can be divided into two divisions, a vocalis division which is important in phonation and an external division which functions in closing the glottis. Fibres in the vocalis are heterogeneous, some stain with mab 4A6, whilst others stain with mabs against limb myosin heavy chains. Fibres in the external division stain almost homogeneous with mab 4A6. The immunohistochemical staining pattern in the cricothyroid muscle resembled that of fast limb muscle: no fibres stained with mab 4A6. Thus, the high speed of contraction of the thyroarytenoid is associated with the same myosin heavy chain found in extraocular muscles, this characteristic is presumably an evolutionary adaptation for rapid closure of the glottis to enhance airway defense mechanisms. PMID- 7499478 TI - Cloning of the cDNA encoding human nonmuscle myosin heavy chain-B and analysis of human tissues with isoform-specific antibodies. AB - Previously, we reported the sequence of cDNA clones encoding amino acids 63 through 723 of the human nonmuscle myosin heavy chain-B isoform. In this paper, we present the derived sequence of the remaining 1303 amino acids along with 5' and 3' untranslated sequences. We made use of the differences between the derived nonmuscle myosin heavy chain-A and -B amino acid sequences to raise isoform specific antibodies. Immunoblot analysis reveals a differential expression of both myosin heavy chain isoforms in a variety of human adult and foetal tissues and cells. When extracts of human adult aorta were subjected to gel electrophoresis, two distinct Coomassie Blue-stained bands and a fused band were seen migrating at approximately 200 kDa. These bands can be detected with four different specific antibodies recognizing the two different smooth muscle myosin heavy chain isoforms (204 kDa and 200 kDa) and the two different nonmuscle myosin heavy chain isoforms (A and B). Using immunohistochemistry, we confirmed the presence of the four different isoforms in adult and foetal aortas. PMID- 7499480 TI - Reversible vacuolation of the transverse tubules of frog skeletal muscle: a confocal fluorescence microscopy study. AB - A confocal microscope was used to investigate the reversible vacuolation of frog skeletal muscle fibres produced by the efflux and entry of glycerol (80-100 mM). The formation, development and disappearance of vacuoles was observed in the fibres by staining simultaneously with two fluorescent membrane probes, RH414 and DiOC6(3). The styryl dye, RH414, stains only the plasmalemma and the membranes of the transverse tubules. In normal and glycerol-loaded fibres, RH414 revealed regular, narrow dotted bands located at the position of the Z-lines. Glycerol removal produced, within 2-10 min, many empty round vacuoles (0.4-1.5 microns in diameter) that were apparently anchored to the stained bands. Later on, individual vacuoles tended to enlarge and align into longitudinal chains of vacuoles. Neighbouring vacuoles that contacted each other fused to form large vacuoles up to several sarcomeres in length. Neither the T-tubules, nor the vacuoles, were stained by DiOC6(3). However, glycerol efflux was also accompanied by a redistribution of sarcoplasmic reticulum membranes and by changes in mitochondria that were revealed on staining the same fibres with the carbocyanine dye, DiOC6(3). The alterations in staining patterns revealed by RH414 and DiOC6(3) were completely reversible. Within 5-10 min after a second application of glycerol, the pattern of staining returned to normal with the exception of very bright, spots stained with RH414, which appeared in place of many but not all of the vacuoles, and probably correspond to the irregular nets of T-tubules observed under the electron microscope in such fibres. They are considered to be defects in regeneration of the T-system after vacuolation. The vacuolation/devacuolation cycle could be repeated several times following glycerol efflux and entry. The development and disappearance of vacuoles then mainly involved conversion of bright spots to large vacuoles and vice versa. Some possible mechanisms of vacuole formation and disappearance are discussed, and it is suggested that vacuolation of the T-system may be important in relation to regulating the volume of skeletal muscle cells. PMID- 7499479 TI - Inositol 1,4,5-trisphosphate receptor in skeletal muscle: differential expression in myofibres. AB - The role of inositol 1,4,5-trisphosphate as a second messenger in signal transduction has been well established in many cell types. However, conflicting reports have led to a controversy regarding the role, if any, of inositol 1,4,5 trisphosphate signalling in skeletal muscle. Indeed, expression of the inositol 1,4,5-trisphosphate receptor has not previously been demonstrated in skeletal muscle. In the present study we used in situ hybridization, immunohistochemistry, and [3H]-inositol 1,4,5-trisphosphate binding to demonstrate that rat skeletal muscle fibres contain inositol 1,4,5-trisphosphate receptors. RNAse protection and partial sequencing suggested that the inositol 1,4,5-trisphosphate receptors expressed in skeletal muscle was most similar to the non-neuronal form of the type 1 inositol 1,4,5-trisphosphate receptor. While in situ hybridization showed inositol 1,4,5-trisphosphate receptor mRNA in all types of skeletal myofibres, immunodetectable inositol 1,4,5-trisphosphate receptor protein and specific [3H] inositol 1,4,5-trisphosphate binding sites were preferentially expressed in slow oxidative (type I) and fast oxidative-glycolytic (type IIA) fibres, but not in fast glycolytic (type IIB) fibres. These findings indicate that an inositol 1,4,5 trisphosphate receptor is preferentially expressed in oxidative fibres of skeletal muscle. PMID- 7499481 TI - Pressure-induced changes in the isometric contractions of single intact frog muscle fibres at low temperatures. AB - Effects of increased hydrostatic pressure (range 0.1-10 MPa) on isometric twitch and tetanic contractions of single intact muscle fibres, isolated from frog tibialis anterior muscle, were examined at 4-12 degrees C. The tension changes produced on exposure to steady high pressures are compared with those produced on exposure to low concentrations of caffeine (0.5 mM, subthreshold for contracture) and when pressure is rapidly released during a contraction. The peak twitch tension was potentiated by pressure accompanied by increased rate of tension rise and increased duration; the pressure sensitivity of twitch tension was approximately 8% MPa-1. The correlation between the rate of tension rise and peak tension in caffeine-induced twitch tension potentiation was quantitatively similar to that in pressure-induced twitch potentiation. Experiments involving the rapid release of pressure (approximately 2 ms) during twitch contractions demonstrate that high pressure need only be maintained for a brief period during the early part of tension development to elicit full twitch potentiation. The tetanic tension was depressed by pressure (approximately 1% MPa-1). Results demonstrate that the major effect of increased hydrostatic pressure on intact muscle fibres, which results in tension potentiation, is complete very early during contraction and is similar to that of caffeine. PMID- 7499482 TI - Fast and slow rat muscles degenerate and regenerate differently after whole crush injury. AB - The whole-crush injured rat skeletal muscle was used as a model to explore the regenerating potentialities of fast and slow muscles. Laminin was chosen to follow changes in basal lamina and desmin to visualize new muscular elements; they were revealed by immunofluorescence on cryostat sections of either fast (extensor digitorum longus) or slow (soleus) regenerating muscle. Soleus myolysis was rapid, extensive and heterogeneous. Basal laminae were nearly destroyed. In contrast, extensor digitorum longus maintained its basal lamina framework during myolysis. Soleus reconstruction began early, following the pattern of remaining basal laminae as closely as possible, but regeneration stagnated from day 16 and the regenerated muscle was fibrotic. In extensor digitorum longus, reconstruction progressed slower than in soleus, but regularly from the periphery toward the centre of the muscle. The regenerated extensor digitorum longus showed a quasi normal structure from day 16. At the end of the process, the elimination of old basal lamina was completed in extensor digitorum longus, but was not achieved in soleus. We propose that the old basal lamina should help the initiation of reconstruction. This new model also underlines the importance of the turnover of basal laminae in muscular regeneration, and will be useful to understand the background of the different regenerative response of both muscles. PMID- 7499484 TI - Calcium sensitivity modulation of myofilaments in cardiac and smooth muscle. A meeting to celebrate Professor Johann Caspar Ruegg's 65th birthday. Heidelburg, 19-22 March 1995. Abstracts. PMID- 7499485 TI - Gamma delta T cell receptor variable region usage during the development of experimental allergic encephalomyelitis. AB - The molecular diversity of gamma delta T cells has not previously been investigated in experimental allergic encephalomyelitis (EAE). This study characterised the gamma delta T cell receptor (TCR) variable (V) region repertoires of T cells infiltrating the brains of EAE mice during development of the disease. TCR gamma- and delta-specific cDNAs were synthesised from total RNA prepared from brain samples and transcription of rearranged V genes was assessed by polymerase chain reaction amplification of TCR V-C transcripts and Southern blot analysis. In the early stages of EAE, the TCR gamma-chain repertoire consisted of V gamma 1-3 and V gamma 6 transcripts and, similarly, a few V delta transcripts that used primarily V delta 1, V delta 4 and V delta 5 gene segments were detected. During the progression of EAE, however, most V gamma and V delta TCR transcripts were observed in the brain. These results indicate that in the course of murine EAE there is an initial infiltration into the brain of a restricted population of gamma delta T cells followed by a heterogeneous gamma delta TCR repertoire as the disease develops. Moreover, the data suggest that gamma delta T cells may play a role in the pathogenesis of demyelinating autoimmune disease. PMID- 7499483 TI - Na channel density in extrajunctional sarcolemma of fast and slow twitch mouse skeletal muscle fibres: functional implications and plasticity after fast motoneuron transplantation on to a slow muscle. AB - Na channel densities were measured in fast and slow twitch mouse skeletal muscle fibres using the loose patch voltage clamp technique. It was found that Na channel density was approximately four times greater in fast twitch fibres than in slow. Computer simulations of action potential propagation in these fibres strongly suggest that the higher channel densities in fast twitch fibres are necessary to maintain action potential amplitude and fidelity of transmission across the neuromuscular junction, especially during the periods of rapid stimulation that these fibres are subjected to by their motoneurons. Transplantation of a foreign nerve containing axons which had previously innervated fast twitch fibres on to a slow twitch muscle resulted in an approximate doubling of the Na channel density in fibres innervated by the foreign nerve. These results suggest that motoneurons may exert considerable control over Na channel density in the muscle fibres they innervate. PMID- 7499486 TI - Expression of the inducible isoform of nitric oxide synthase in the central nervous system of mice correlates with the severity of actively induced experimental allergic encephalomyelitis. AB - A cytokine-mediated excessive increase in nitric oxide (NO) by macrophages or glial cells via an inducible isoform of NO synthase (iNOS) has been proposed to play an important role in demyelinating diseases. To further investigate the role of iNOS in demyelination, experimental allergic encephalomyelitis (EAE), a known animal model of multiple sclerosis (MS) in mice, was chosen in this study. A semiquantitative reverse transcriptase-polymerase chain reaction (RT/PCR) analysis revealed an increase in the mRNA levels of iNOS and cytokines known to induce iNOS or inflammatory cytokines (interleukin (IL)-1 alpha, IL-1 beta, IL-2, IL-6, interferon (IFN)-gamma, tumor necrosis factor (TNF)-alpha and TNF-beta) in the spinal cord corresponding to the severity of the disease without significant change in the mRNA levels of immunoregulatory cytokines (IL-4, IL-10 and transforming growth factor (TGF)-beta) during the course of EAE. An immunohistochemical examination of the spinal cord using an iNOS-specific antibody showed iNOS-positive cells to be mainly inflammatory cells with a higher frequency of iNOS-positive cells at the peak of EAE than in the early phase. These iNOS-positive cells at the peak appeared to be composed of infiltrating macrophages and most of them were located in the necrotic area. These results suggested that cytokine-induced excessive NO via iNOS by macrophages caused tissue damage in the central nervous system in EAE. PMID- 7499487 TI - Sequence of kappa-opioid receptor cDNA in the R1.1 thymoma cell line. AB - To our knowledge, this is the first demonstration of kappa-opioid receptor mRNA in cells of the immune system. While the presence of opioid receptors on cells of the immune system has been controversial, cell-binding analysis has indicated that the kappa-opioid receptor is expressed by the immature T cell line R1.1. We have developed a reverse transcriptase-polymerase chain reaction protocol to amplify the mRNA extracted from R1.1 cells with primers derived from the cDNA sequence of the mouse kappa-opioid receptor. Nucleotide sequences of the amplified products were examined and two populations of cDNA were detected which differ in the 5' region upstream of the ATG start codon. Comparison of these sequences to the previously published kappa-opioid receptor cDNA sequence suggests the presence of an intron-exon junction in the 5' non-coding region. PMID- 7499488 TI - Cerebrospinal fluid free kappa light chains versus IgG findings in neurological disorders: qualitative and quantitative measurements. AB - In this study free kappa light chains in cerebrospinal fluid (CSF) were determined both by an affinity mediated capillary blotting technique after isoelectric focusing (IEF) in agarose gel and by a quantitative enzyme linked immunosorbent assay (ELISA). The free kappa results were compared with the IgG findings in 4 neurological patient groups with a distinct CSF IgG pattern: (1) CSF without oligoclonal IgG bands, (2) CSF with serum derived IgG bands, (3) CSF restricted IgG bands and (4) CSF restricted and serum derived IgG bands. Oligoclonal free kappa bands are nearly absent in CSF of groups 1 + 2, and present in 88% of group 3 and 84% of group 4 patients. We could also establish free kappa indices from specimens in the 4 groups in analogy to IgG indices. Group 1 had a median free kappa index of 1.1, group 2: 1.0 and groups 3 + 4: 10.0. The correspondence between immunoblot and index findings for free kappa is better than for IgG. Free kappa index is more sensitive but somewhat less specific than IgG index for establishing intrathecal immune production. PMID- 7499489 TI - Increased activity of hypothalamic corticotropin-releasing hormone neurons in multiple sclerosis. AB - Clinical observations and animal studies suggest that the hypothalamo-pituitary adrenal (HPA) axis plays a role in the susceptibility to and the recovery from multiple sclerosis (MS). Since the HPA-axis is under the control of corticotropin releasing hormone (CRH) neurons of the hypothalamus, we determined 2 parameters for activation of the CRH neurons in the hypothalamic paraventricular nucleus (PVN) in MS patients. Since the HPA-axis is more activated in MS, we expected an increased activity of CRH neurons. We also expected to see an age-related increase in CRH activity, because of the possible role of the HPA-axis in the age related decrease in susceptibility to MS. The number of CRH cell profiles and the proportion of CRH neurons co-expressing vasopressin were used as parameters for activity. CRH cell population became more activated both in control and MS patients, from 40 years of age onwards, when the prevalence of MS starts to decrease in the population. The CRH neurons showed a significantly higher level of activation in MS patients than in controls, as appeared from the 3-fold increase in CRH cell number and the 4.5-fold increase in cells co-expressing CRH and vasopressin (AVP). PMID- 7499490 TI - Virus-activated T cells regulate expression of adhesion molecules on endothelial cells in sites of infection. AB - To study the role of cell adhesion molecules in the fatal CD8+ T-cell mediated meningitis which is induced by intracerebral infection with lymphocytic choriomeningitis virus, the expression of relevant molecules on inflammatory cells and local endothelium was analyzed immunohistochemically. Most inflammatory cells were strongly positive for LFA-1, VLA-4, Pgp-1 and ICAM-1. Expression of ICAM-1 and VCAM-1 was upregulated on the endothelial cells in immunocompetent mice, but not in T-cell deficient nude mice. Analysis of mice deficient in either CD4+ or CD8+ T cells, revealed that not only was the inflammatory reaction dependent on the presence of CD8+ cells, but these cells also appeared to be required for maximal upregulation of ICAM-1 and VCAM-1 on the endothelial cells. These results indicate that virus-specific CD8+ T cells are crucially involved in regulating the inflammatory reaction through effects on endothelial expression of adhesion molecules. PMID- 7499491 TI - Human microglia mediate anti-Cryptococcus neoformans activity in the presence of specific antibody. AB - The interaction of the opportunistic fungus Cryptococcus neoformans with human microglia was studied in vitro in the presence and absence of capsule binding antibody. In the absence of capsule binding antibody there was little or no phagocytosis. Addition of the murine monoclonal antibody (mAb) 2H1 (IgG1, kappa) to the capsular glucuronoxylomannan (GXM) produced a dose-dependent enhancement of C. neoformans phagocytosis by microglia. Phagocytosis resulted in marked inhibition of fungal proliferation. Microglial antifungal activity was studied by colony forming unit assay, L-[3H]leucine incorporation assay, and phase contrast microscopy. At microglia: C. neoformans ratios of 10:1 to 80:1 fungal growth was reduced by 61-95%. Inhibitors of nitric oxide synthase and reactive oxygen intermediates did not prevent antifungal activity mediated by human microglia. Transmission electron microscopic studies revealed that although some internalized yeast cells were killed, the majority were intact consistent with fungistasis. Human microglia cells are potent effector cells against C. neoformans in vitro in the presence of specific antibody. Enhancement of microglial activity in vivo by opsonins may be a useful therapeutic strategy. PMID- 7499493 TI - Inducible preprotachykinin mRNA expression in mucosal lymphoid organs following oral immunization with Salmonella. AB - Using a sensitive and specific reverse transcribed-polymerase chain reaction (RT PCR), it was possible to quantify relative increases in preprotachykinin (PPT) mRNA expression in vivo following oral administration of Salmonella. Despite the presence of constitutive levels of PPT mRNA expression in the Peyer's patches, expression of this mRNA increased within 20 h following oral administration of Salmonella. Increases in PPT mRNA expression were also detected in both the mesenteric lymph nodes and spleens following Salmonella administration despite the lack of constitutive PPT mRNA expression in these lymphoid organs. Furthermore, mononuclear leukocytes contributed to this increased expression of PPT mRNA, suggesting that the initial immune response against Salmonella occurs in the presence of increased tachykinin expression. PMID- 7499492 TI - HLA-class II alleles in Guillain-Barre syndrome and Miller Fisher syndrome and their association with preceding Campylobacter jejuni infection. AB - HLA typing for class II alleles was performed on 97 patients with Guillain-Barre syndrome or Miller Fisher syndrome and compared with 100 controls. There was a significant association between HLA-DQB1*03 and preceding Campylobacter jejuni infection (Pc = 0.05). PMID- 7499494 TI - Suppression of hyperacute and passively transferred experimental autoimmune encephalomyelitis by the anti-oxidant, butylated hydroxyanisole. AB - Butylated hydroxyanisole (BHA) was used to treat hyperacute, ordinary passive, and hyperacute passive experimental autoimmune encephalomyelitis (EAE) in the Lewis rat. The anti-oxidant, delivered via mini-osmotic pumps, reduced the incidence, severity and mortality in hyperacute EAE and also reduced the incidence, severity and duration of disease in passively induced EAE and hyperacute passive EAE. In all cases, cellular infiltration by both mononuclear and polymorphonuclear leukocytes were significantly reduced in treated rats. BHA appears therefore to act at the effector stage of EAE, reducing cellular infiltration in the brain and spinal cord and minimising clinical signs without blocking sensitisation or activation. This was supported by the finding that spleen cells from BHA-treated donors immunised for hyperacute EAE transferred disease at least as well as cells recovered from untreated donors. PMID- 7499496 TI - Thymic vasopressin (AVP) transgene expression in rats: a model for the study of thymic AVP hyper-expression in T cell differentiation. AB - The peptide arginine vasopressin (AVP) is present within tissues of the immune system and has been implicated in T cell differentiation. We have investigated the expression and production of AVP in the thymus of rats which carry a rat AVP transgene. A 100% increase in thymic AVP immunoreactivity (ir) was detected in transgenic (TG) animals compared to age-matched wild-type (WT) controls. When tissues from TG and WT thymuses were subjected to reversed-phase high-performance liquid chromatography, ir-AVP eluted as a single peak which co-eluted with the standard. Immunocytochemical staining identified the presence of AVP in large epithelial cells within the thymic cortex in both WT and TG animals. The AVP precursor product neurophysin was also detected in epithelial cells in WT and TG thymuses. In situ hybridisation histochemistry using a probe specific for transgenic AVP mRNA revealed that the AVP transgene was expressed in TG thymic cells with a similar morphology and distribution to those which expressed endogenous AVP peptide in WT animals. These results demonstrate that the cellular location and immunoreactive form of AVP expressed in TG animals are similar to that found in WT controls. Thus the TG rat appears to be a model of true physiological, rather than ectopic, over-expression of AVP in the thymus. The hyper-expression of AVP in the thymic epithelial cells of TG animals provides a model in which can be studied the influence of AVP on T cell development and differentiation within the thymus. PMID- 7499495 TI - Thymomas express epitopes shared by the ryanodine receptor. AB - Myasthenia gravis (MG) patients with thymoma have antibodies against ryanodine receptor (RyR) of skeletal and heart muscle. In this study, thymomas were examined for reactivity with a panel of polyclonal rabbit antibodies against various short peptides of RyR. An antibody against peptide C2 in the transmembrane region of RyR stained thymoma epithelial cells in cryosections of 17/23 thymomas, and detected a 40-kDa peptide in Western blotting of a thymoma membrane fraction. The other RyR antibodies did not react with thymoma tissue. The anti-C2 RyR antibody did not react with normal thymus, tonsil or carcinoma of colon. The results strongly indicate that epithelial thymoma cells express an epitope shared by the transmembrane region of skeletal and cardiac muscle RyR. PMID- 7499497 TI - Modulatory effects of [Met5]-enkephalin on interleukin-1 beta secretion from microglia in mixed brain cell cultures. AB - In the present study, functional interactions between [Met5]-enkephalin (ME), naloxone and lipopolysaccharide (LPS) on interleukin-1 beta (IL-1 beta) immunostaining and secretion have been assessed in mixed brain cell cultures from embryonic day 17 mice. Adding ME alone or together with LPS to the culture increased the release of IL-1 beta after 48 h in a concentration-dependent fashion. In situ hybridization studies showed that LPS, but not ME, increased the abundance of IL-1 beta mRNA. The enhanced release of IL-1 beta caused by ME or LPS was partially blocked by naloxone. LPS induced concentration-dependent morphological changes in microglia in mixed brain cell cultures, identified by a monoclonal antibody F4/80 which is specific for macrophages/microglia. Despite increasing IL-1 beta release into the media, ME (10(-8) M) did not induce morphological changes in microglia. Naloxone alone also had no effect on glial morphology; however, the LPS-induced morphological changes were blocked by naloxone. Our data indicate that both exogenous and endogenous opioids regulate IL-1 beta production by microglial cells in the mixed brain cell cultures. PMID- 7499499 TI - Characteristics of cloned cerebrovascular endothelial cells following infection with Theiler's virus. I. Acute infection. AB - The present study describes the replication of Theiler's virus in cloned cerebrovascular endothelial cells (CVE) isolated from strains of mice that are either susceptible or resistant to Theiler's virus-induced demyelination (TVID). CVE isolated from all strains of mice were equally permissive to Theiler's virus infection. Interferon-gamma and tumor necrosis factor-alpha were found to inhibit the replication of Theiler's virus in CVE. A correlation between susceptibility to demyelination and the ability of Theiler's virus to induce MHC Class I on CVE was demonstrated. PMID- 7499498 TI - Release of myelin basic protein-degrading proteolytic activity from microglia and macrophages after infection with Theiler's murine encephalomyelitis virus: comparison between susceptible and resistant mice. AB - Theiler's murine encephalomyelitis virus (TMEV) produces a chronic inflammatory demyelinating disease in its natural host, the mouse. A delayed-type hypersensitivity (DTH) response to viral antigens generally correlates with susceptibility to the disease and is thought to play an important role in the pathogenesis of demyelination in this model of human multiple sclerosis (MS). The hallmark of DTH responses is the recruitment by activated Th-1 cells of lymphoid cells and especially macrophages in infected areas. It is believed that soluble factors released by these cells would produce tissue damage, particularly myelin breakdown. In the present study, we compared TMEV-infected macrophages and microglia, isolated from both susceptible SJL/J and resistant C57BL/6 mice, for their ability to secrete proteolytic enzymes capable of degrading myelin basic protein. In addition, we studied whether supernatants from infected microglia/macrophages were also capable of killing oligodendrocytes in the same in vitro system. As detected by SDS-PAGE, MBP-degrading proteolytic activity was found only in supernatants from infected SJL/J microglia and macrophages, but not in supernatants collected from infected C57BL/6 microglia and macrophages, or in supernatants from mock-infected SJL/J and C57BL/6 cells. Similarly, incubation of E20.1 cells, an immortalized line of oligodendrocytes, with infected SJL/J, but not C57BL/6 supernatants, resulted in cytotoxic activity. When cells from resistant C57BL/6 mice were treated with LPS, they became susceptible to infection and also secreted proteolytic enzymes. The proteolytic activity released from infected microglia and macrophages was found to be dose-dependent, was inactivated by heat, and was inhibited by phenylmethylsulphonyl fluoride (PMSF). These results indicate that a serine protease is released from infected microglia and macrophages and suggest a role for proteases in TMEV-induced myelin injury. PMID- 7499500 TI - Characteristics of cloned cerebrovascular endothelial cells following infection with Theiler's virus. II. Persistent infection. AB - Cloned cerebrovascular endothelial cells (CVE) persistently infected with Theiler's virus (PI-CVE) have been established and characterized. The CVE were derived from strains of mice that are susceptible (SJL/J and CBA) and resistant (BALB/c) to Theiler's virus-induced demyelination (TVID). The cells were persistently infected with either the BeAn or GDVII strains of Theiler's virus in vitro and studied at various passage levels for infectious virus, viral antigen and the expression of major histocompatibility complex (MHC) Class I and II antigens. The virus replicated to lower titers than in acutely infected CVE and appeared to be more cell-associated. Flow cytometric analysis revealed that 18 39% of the PI-CVE contained viral antigen. Persistently infected CVE derived from SJL/J and CBA mice expressed high levels of MHC Class I, whereas BALB/c PI-CVE did not. MHC Class II was upregulated by IFN-gamma in SJL/J PI-CVE albeit at a slightly lower level than in uninfected CVE. In addition, the PI-CVE demonstrated increased levels of mRNA for IL-1 beta when compared to uninfected CVE. PMID- 7499501 TI - Pituitary adenylate cyclase-activating polypeptides (PACAP27 and PACAP38) inhibit the mobility of murine thymocytes and splenic lymphocytes: comparison with VIP and implication of cAMP. AB - In the present study, the effects of PACAP27, PACAP38 and VIP in a concentration range from 10(-13) to 10(-6) M were studied in vitro on the spontaneous and directed mobility of lymphocytes from rat spleen and thymus. The results show that VIP and both PACAPs inhibit significantly and in a similar way the mobility of lymphocytes from thymus and spleen, and the maximal effects were observed at 10(-9) M and 10(-8) M. The three neuropeptides significantly increased cAMP concentrations. Moreover, incubation with increasing PMA concentrations showed a progressive enhancement of chemotaxis of lymphocytes, which was partially prevented by VIP, and both PACAPs. Incubation with forskolin caused decrease in the chemotaxis of thymocytes and splenocytes, and the presence of VIP or PACAP peptides was not synergistic in the inhibitory effect on lymphocyte chemotaxis, suggesting that the three neuropeptides and forskolin mediate their actions by the same intracellular pathway. This study showed the ability of the VIP receptor antagonist (N-Ac-Tyr1,D-Phe2)-GRF(1-29)-NH2 to partially reverse the inhibitory effect of both PACAPs and VIP on chemotaxis, suggesting that PACAP receptors are identical or very similar to VIP receptors in both thymocytes and splenocytes. These data suggest that PACAP27 and PACAP38 can be included as two novel immunoregulatory peptides that can modulate cell mobility on central and peripheral lymphoid organs. PMID- 7499502 TI - The effects of citrullination or variable amino-terminus acylation on the encephalitogenicity of human myelin basic protein in the PL/J mouse. AB - The post-translational modifications of myelin basic protein (MBP) in the form of citrullination and varying length of amino-terminus acylation may modify the biological functions and immunological features of MBP. Both modifications influence the reaction of antibodies and specific T cells recognizing MBP. The present study was undertaken to compare the encephalitogenicity of the citrullinated isomer of MBP (MBP-C8) with the unmodified isomer of MBP (MBP-C1) and to determine if the length of amino-terminal acylation of MBP peptide 1-21 altered an encephalitogenic epitope. MBP-C8, whether from patients with or without multiple sclerosis (MS), and MBP-C1 could induce active experimental allergic encephalomyelitis (EAE) in PL/J mice. A trend of reduced severity of EAE was observed in MBP-C8-injected animals. An increase in the length of amino terminus fatty acid decreased the encephalitogenicity of MBP peptide 1-21 for both active and adoptive EAE in PL/J mice. Only lymph node cells sensitive to MBP peptide acetyl 1-21 and butyl 1-21 could transfer clinical EAE. In adoptive EAE, MBP peptides hexyl and octyl 1-21 induced moderate histopathological but no clinical change, whereas MBP peptide decyl 1-21 caused neither. A broadening in the antibody response could be detected in the sera of mice with active EAE induced by MBP-acylated peptides 1-21. Our findings demonstrate that encephalitogenicity is retained in the presence of citrullination but that the length of amino-terminus acylation diminishes the encephalitogenicity of MBP in the PL/J mouse.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7499503 TI - Both anti-CD11a (LFA-1) and anti-CD11b (MAC-1) therapy delay the onset and diminish the severity of experimental autoimmune encephalomyelitis. AB - Experimental autoimmune encephalomyelitis (EAE) is a demyelinating disease of the central nervous system (CNS) induced in rodents by activated CD4+ T cells specific for various myelin proteins such as myelin basic protein and proteolipid protein. The disease is characterized by breach of the blood-brain barrier, perivascular infiltration of leukocytes into the CNS, local inflammation and demyelination in the form of plaques. In this study, we evaluated the effect of administration of antibodies to two members of the beta 2 integrin sub-family of adhesion molecules, CD11a and CD11b, on the onset and progression of EAE. CD11a and CD11b are involved in cell-cell interactions leading to T cell and macrophage extravasation to inflammatory sites and T cell activation. Our results show that anti-CD11a antibodies could completely block the induction of EAE and anti-CD11b antibodies significantly delayed the onset and diminished the severity of clinical signs of EAE even when injections were initiated at the first appearance of clinical signs. PMID- 7499504 TI - Expression of mRNA for 55-kDa and 75-kDa tumor necrosis factor (TNF) receptors in mouse cerebrovascular endothelium: effects of interleukin-1 beta, interferon gamma and TNF-alpha on cultured cells. AB - The interactions between tumor necrosis factor-alpha (TNF-alpha) and its receptors have been implicated to play an important role in the pathogenesis of experimental autoimmune encephalomyelitis (EAE). The effects of cytokines on the steady state levels of TNF receptor (TNFr) mRNA in cerebrovascular endothelial cells (CVE) cultured from EAE-susceptible (SJL/J) and EAE-resistant (BALB/c) mice were examined. Interferon-gamma and interleukin-1 beta up-regulated the levels of both the 55-kDa and 75-kDa TNFr mRNA, while TNF-alpha had no effect. No differences were observed in cytokine-induced TNFr mRNA levels between BALB/c and SJL/J CVE that might explain differences in EAE susceptibility. PMID- 7499505 TI - Interferon-gamma induces T lymphocyte proliferation in multiple sclerosis via a Ca(2+)-dependent mechanism. AB - The intracellular mechanisms underlying T lymphocyte activation leading to demyelination in multiple sclerosis (MS) have not yet been clarified. We have recently reported that interferon (IFN)-gamma activates a novel trans-plasmalemma Ca2+ influx on T lymphocytes (mainly CD4+) from patients with MS which induces intracellular Ca2+ ([Ca2+]i) elevation. Since Ca2+ is an essential second messenger in regulating transcription of T lymphocyte activation genes, we have evaluated how [Ca2+]i elevation due to the activity of this particular influx affects T lymphocyte proliferative behaviour in 12 influx-positive relapsing remitting MS (RR-MS) patients. Fourteen influx-negative RR-MS patients and 14 healthy donors were used as controls. In lymphocytes from healthy controls, a significant correlation (r = 0.62; P < 0.001) was found between [Ca2+]i levels and proliferation rate after phytohemagglutinin (PHA) stimulation. Sustained proliferation was induced in T lymphocytes by > or = 10 micrograms/ml of PHA, a dose leading to a [Ca2+]i increase of at least 45% over basal level. Similar [Ca2+]i elevations were obtained when > or = 10 micrograms/ml of PHA were used on cells from RR-MS patients. However, T lymphocytes from RR-MS patients, but not from healthy donors, proliferated also in response to 1 micrograms/ml of PHA, indicating a state of preactivation. Moreover, 1 microgram/ml of PHA used in combination with suboptimal doses of IFN-gamma (5 UI/ml) doubled the proliferation rate of influx-positive MS cells, but not influx-negative MS cells or cells from healthy donors compared to the values obtained using PHA alone (P < 0.01).(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7499506 TI - Circulating natural killer cells but not cytotoxic T lymphocytes are reduced in patients with active relapsing multiple sclerosis and little clinical disability as compared to controls. AB - Triple antibody flow cytometry was used to compare the populations of CD56+ effector cells in the peripheral circulation of 29 patients with relapsing multiple sclerosis (MS) and little disability who were exacerbation-free for over 2 months and 29 healthy control subjects. Populations were characterized by two panels of antibodies (CD8, CD16, CD56 and CD3, CD8, CD56), as well as by size or granularity. In the MS patients, mature natural killer (NK) cells (CD3-CD8-CD56+) of small size and low granularity were significantly reduced compared to normals (P(0.0003). The quantities of other effector cells (cytotoxic T lymphocytes, large granular lymphocytes and monocytes) were not different in MS patients compared to the control subjects. Also, we identified a previously unrecognized population of CD56+ monocytes (CD3-CD14+CD56+) in both the normal control subjects and the MS patients which would have been misclassified as NK cells using one or two antibody cytometry employed in previous studies. PMID- 7499508 TI - Increased antibody production in peripheral blood mononuclear cells after plasma exchange therapy in multiple sclerosis. AB - It has been difficult to study antibody synthesis in vivo following plasma exchange (PE) because of the intra- and extravascular compartmentation of immunoglobulins, and their altered lymphatic flow and catabolism after the procedure. There is fragmentary experimental and clinical evidence that antibody synthesis may increase due to removal of autoregulatory factors by PE. The present study showed increased in vitro spontaneous immunoglobulin (Ig) G and IgM production, as well as increased spontaneous lymphocytic proliferation, after intensive PE in 10 multiple sclerosis (MS) patients. Cytofluorometric control of cultured mononuclear cell populations revealed no change with treatment except for a small increase in B lymphocytes inadequate to account for the increased immunoglobulin production. These findings indicate an activation of the immune system through PE, and suggest the possible need for adjunctive immunosuppression to control antibody production in autoimmune diseases with subacute, relapsing, or chronic courses when PE is employed as a therapeutic modality. PMID- 7499510 TI - Let's take on the real dragon: profiteering in health care. PMID- 7499509 TI - Macrophages and microglia in HSV-1 infected mouse brain. AB - We have examined the macrophage and microglial cell response in mouse brains during the course of acute herpes simplex encephalitis produced by intranasal inoculation of virus. Macrophage infiltration and a microglial cell reaction were detected from 24 h after initial detection of HSV-1 antigen in brain. Macrophage infiltration was confined to focal areas of infection but the microglial cell reaction occurred in a more widespread distribution. Following disappearance of viral antigen, with resolution of the acute infection, macrophage infiltration and a focal microglial cell reaction remained for several weeks, providing a marker of sites of previous infection. PMID- 7499507 TI - Anti-idiotypic antibodies: a useful alternative for studying the biochemical expression of mu/delta opioid binding sites in mammalian brain. AB - A polyclonal antiserum was produced against opioid binding sites using an anti idiotypic approach whereby antibodies directed against the opioid agonist DSLET were used as immunogen. The anti-idiotypic antiserum recognized specific brain proteins with molecular masses of 76 +/- 4, 73 +/- 4 and 59 +/- 3 kDa, respectively. The immunolabeling of these proteins was mainly inhibited by mu, delta opioid agonists and a general antagonist, naloxone. The inhibition of immunoprecipitation by opioid agonists and antagonist and the developmental expression of these immunoreactive proteins found to occur during brain ontogeny strongly suggest that these three proteins were mu, delta but not kappa opioid binding sites. The anti-idiotypic antiserum both inhibits 3H-DADLE binding and mimics the inhibitory agonist effects on the stimulated cAMP level of NG 108-15 cells which expressed delta opiate receptors. Numerous mammalian brain opioid binding sites were labeled, due to the fact that the binding site was the epitope recognized by the anti-idiotypic antibodies. From the numerous studies performed with a view to characterizing the specificity of the anti-idiotypic antibodies, it was strongly suggested that the anti-idiotypic antibodies specifically recognize mu/delta opioid binding sites and they can therefore be powerful tools for studying the biochemical expression of these opioid binding sites in mammalian brains. PMID- 7499511 TI - Fashioning a work-based strategy for welfare reform based on international human rights doctrine. AB - The role of work in debates over welfare reform in the United States is analyzed. Five issues are identified and discussed under this heading, with special emphasis on the question of whether enough jobs exist to make it possible for all able-bodied welfare recipients to find paid employment in the regular labor market. It is argued that there are not enough jobs available--not even low-wage jobs--to make this a reasonable goal, and that neither conservatives nor liberals have faced up to the dilemma posed by this job shortage. International human rights doctrine concerning the availability of work at decent wages is cited in support of the proposition that the government should be held accountable for filling this job gap, not only for welfare recipients but for all other job seekers as well. The costs of a hypothetical jobs program capable of achieving this goal are assessed, with the surprising result that it might actually save the public money. It is suggested that such a program could constitute an effective alternative to current welfare reform proposals. PMID- 7499513 TI - The public health leadership institute: leadership training for state and local health officers. AB - This paper describes the Public Health Leadership Institute (PHLI), a program designed to expand and enhance the leadership skills of senior public health officials. The background of PHLI is discussed, and organizations responsible for its organization, governance and funding are revealed. Methods of selecting a cohort of scholars to participate each year, along with the characteristics of scholars selected, are briefly outlined. A calendar of events and description of PHLI curriculum is included. Specific features of the PHLI program, including peer-learning through electronic conferencing, the one-week learning retreat, and the required action-learning projects are described. Also included are results from an evaluation by 99 participants from the first two years of PHLI's existence. The evaluation is based on a survey of the participants' satisfaction with the program and its personal and institutional effects. The Public Health Leadership Institute provides a model of leadership development for senior public health agency officials. The program has been popular with participants, and the vast majority report improved leadership skills. The personal and organizational effects of PHLI address the deficits in leadership identified in the Institute of Medicine's report, The Future of Public Health. PMID- 7499512 TI - Achieving and sustaining full employment. AB - Human rights and public health considerations provide strong support for policies that maximize employment. Ample historical and conceptual evidence supports the feasibility of full employment policies. New factors affecting the labor force, the rate of technological change, and the globalization of economic activity require appropriate policies--international as well as national--but do not invalidate the ability of modern states to apply the measures needed. Among these the most important include: (I) systematic reduction in working time with no loss of income, (2) active labor market policies, (3) use of fiscal and monetary measures to sustain the needed level of aggregate demand, (4) restoration of equal bargaining power between labor and capital, (5) social investment in neglected and outmoded infrastructure, (6) accountability of corporations for decisions to shift or reduce capital investment, (7) major reductions in military spending, to be replaced by socially needed and economically productive expenditures, (8) direct public sector job creation, (9) reform of monetary policy to restore emphasis on minimizing unemployment and promoting full employment. None are without precedent in modern economies. The obstacles are ideological and political. To overcome them will require intellectual clarity and effective advocacy. PMID- 7499514 TI - Alcohol-problem prevention research policy: the need for a phases research model. AB - This paper describes the need and rationale for developing a phases model for guiding alcohol-problem prevention research. A phased approach to prevention research is consistent with such models developed for other health areas including heart disease, cancer, and drug testing. Such a model in alcohol prevention research can provide a means for (1) locating how far research has progressed along a continuum from basic or pre-intervention research to full implementation of preventive action, (2) identifying gaps in research, and (3) determining the level of empirical proof which exists for one or more prevention strategies prior to widespread dissemination. PMID- 7499515 TI - Characteristics of fatal crashes of 16-year-old drivers: implications for licensure policies. AB - Compared with older drivers, and even older teens, greater percentages of 16-year old drivers in fatal crashes were involved in single-vehicle crashes, were responsible for their crashes, were cited for speeding, had high vehicle occupancy (especially other teenagers), and were female. Sixteen-year-olds were less likely than older drivers to have been drinking. In addition, their crashes occurred at different times than those of older drivers, crashes between 10:00 p.m. and 11:59 p.m. on Fridays and Saturdays being especially likely. Information about the characteristics of the crashes of 16-year-olds is important because this is the age at which most states currently allow teenagers to get an unrestricted driver's license. It is also the age at which restrictions on beginning licenses are being considered in some states. The results of this study suggest that restrictions on teenage passengers, and night-driving curfews with pre-midnight starting times--two provisions used in New Zealand's graduated licensing system--would be appropriate in attempts to reduce crashes of beginning 16-year-old drivers, who have the highest fatal crash rate of any single teen age. PMID- 7499516 TI - A randomized trial of transcutaneous extraction atherectomy in femoral arteries: intravascular ultrasound observations. AB - OBJECTIVES: The purpose of this study was to test the hypothesis that in occlusions of the superficial femoral artery, removal of atherosclerotic plaque would result in a higher long-term patency rate compared to balloon dilatation alone. A secondary hypothesis was that long term patency would be proportional to the amount of plaque removed. METHODS: A randomized controlled study of patients with occluded superficial femoral arteries was performed comparing balloon dilatation alone versus a 2.7 mm or a larger (4.0 mm or 4.7 mm) transcutaneous extraction catheter (TEC) atherectomy device followed by balloon dilatation. The effect of these devices on plaque area was assessed directly by intravascular ultrasound imaging. RESULTS: The mean occlusion length was 19.4 cm +/- 11.7 cm. The mean lumen area increased from 4.7 mm2 to 15.1 mm2, primarily due to balloon dilatation, but the mean atheroma area of 19.8 mm2 did not change with either size of TEC device. Although the initial procedure success rate was high (79%), the 6 month patency was only 45%. There was no difference in 6 month patency between the 3 groups. CONCLUSIONS: The data indicate that the TEC atherectomy devices do not remove a significant amount of atherosclerotic plaque in occluded superficial femoral arteries. The 6 month patency is no different with these atherectomy devices than with balloon dilatation alone. The larger (4.0 mm or 4.7 mm) TEC device does not remove any more tissue than the smaller (2.7 mm) device. The use of intravascular ultrasound to quantitate the effects of this atherectomy device provides important insights into the mechanism of action and lack of efficacy of the TEC atherectomy catheter. PMID- 7499517 TI - Degenerated uterine leiomyoma mimicking acute appendicitis: sonographic diagnosis. AB - The present study reviews four cases in which degenerated uterine leiomyomas mimicked appendicitis clinically. The correct diagnosis was made prospectively by sonography in each case. Appendicitis was excluded by graded compression sonography and an exophytic uterine leiomyoma was identified at the point of maximal symptoms. The leiomyomas demonstrated central hypoechoic foci, but no detectable Doppler flow indicating necrosis. Although the phenomenon of a degenerated uterine leiomyoma mimicking appendicitis clinically is known, sonographic diagnosis has not been reported previously. The importance of making this diagnosis sonographically is that management of a degenerated leiomyoma is conservative and unnecessary surgery could be avoided. PMID- 7499518 TI - Hepatic venous Doppler waveforms: changes in pregnancy. AB - A total of 75 fasted healthy normal volunteers were examined during various stages of pregnancy. Examinations were performed in a supine position using an Acuson 128 ultrasound scanner with a 3.5 MHz transducer. Doppler interrogation of the middle hepatic vein was made during quiet respiration by one of three examiners. The Doppler waveforms were subsequently assessed blindly by one of the authors and categorized as normal, damped, or flat. Between 12 and 20 weeks, the majority of patients had normal hepatic pulsatility (64%) with 20% damped and 16% flat. Between 20 and 30 weeks there was a significant change (p > 0.001) with 68% being flat, 20% damped, and only 12% normal. In the last 10 weeks of pregnancy the changes were more marked: 80% were flat, 12% dampened, and 8% normal. There was a profound change in hepatic venous pulsatility during pregnancy. Hepatic waveforms changed from their normal pulsatile nature to become completely flat with increasing gestation. These changes were more frequent and more marked the further gestation progressed. PMID- 7499519 TI - Review of artifacts associated with transrectal ultrasound: understanding, recognition, and prevention of misinterpretation. AB - Artifacts are inadequate representations of the structures being imaged. Transrectal ultrasound (TRUS) used for evaluating rectal tumors has its own, unique spectrum of artifacts such as (1) pseudomasses (beam thickness: imaging of rectal folds; mirror image: reflection at an intraluminal fluid level); (2) inadequate size of the lesion (mirror image or grating lobes); (3) simulation of malignant infiltration (beam thickness, attenuation or refraction); (4) incomplete field of view (shadowing; reverberation or mirror-image); (5) confusing echo patterns (side lobe artifacts or mirror image: reflection at the balloon surface). The understanding of the physical properties of ultrasound is the basis for the recognition of these artifacts and prevention of misinterpretation. We present a review of these artifacts and their causes. PMID- 7499520 TI - Tuberculous myositis: CT and sonographic findings in two cases. PMID- 7499521 TI - Biliary ascariasis: ultrasound diagnosis. PMID- 7499522 TI - Cor triatriatum dexter diagnosed by transesophageal echocardiography. PMID- 7499523 TI - The engorged fallopian tube: a new sonographic sign for adnexal torsion. PMID- 7499524 TI - Developing mossy fiber terminal fields in the rat cerebellar cortex may segregate because of Purkinje cell compartmentation and not competition. AB - Many mossy fiber afferent projections to the rat cerebellar cortex terminate in parasagittal bands. In particular, the anterior lobe vermis of the cerebellum contains alternating bands of mossy fibers from the spinal cord and external cuneate nuclei. The cerebellar cortical efferents, the Purkinje cells, are also organized in parasagittal bands. These can be revealed by immunochemical staining for the antigen zebrin II, which is selectively expressed by bands of Purkinje cells. In some cases, the boundaries between mossy fiber terminal fields align with identified transitions between zebrin+/- sets of Purkinje cells, whereas others are located within apparently homogeneous Purkinje cell compartments. Two theories can explain the terminal-field topography: In one view, mossy fiber terminals segregate during development, because growth cones from different sources compete for common territory. Alternatively, mossy fiber growth cones directly recognize chemically distinct target territories, and activity-dependent mechanisms play only minor roles. To explore these issues, two sets of experiments were performed. First, the terminal-field map of the neonatal spinocerebellar projection was compared to the Purkinje cell compartmentation as revealed by anticalbindin immunocytochemistry. Second, subsets of spinocerebellar mossy fiber afferents were ablated early in postnatal development, and the consequences for the neighboring cuneocerebellar terminal fields were mapped in the adult with reference to the zebrin II+/- compartments. These experiments revealed no evidence that competitive interactions constrain the mossy fiber terminal-field distribution but, rather, suggest that the organization of the mossy fiber projections follows the compartmentation of the Purkinje cells. PMID- 7499525 TI - Compartmental organization of layer IVA in human primary visual cortex. AB - Immunostaining for three neuronal proteins, nonphosphorylated neurofilament protein (with antibody SMI-32), calbindin, and parvalbumin, was used to examine the organization of layer IV in human primary visual cortex (area 17 or V1) specifically to determine whether, similar to the case in macaque V1, layer IVA is present and is divided into neurochemically distinct compartments. All three proteins are expressed by neurons that are unevenly distributed in layer IV of human V1; immunostaining for each protein includes a thin band corresponding to layer IVA of classic cytoarchitectonic studies. In this band, nonphosphorylated neurofilament protein immunoreactivity is present in relatively broad clusters of pyramidal cell somata and dendrites that appear as upwardly protruding parts of intense immunostaining in layer IVB, whereas immunoreactivity for calbindin and parvalbumin exists in somata of nonpyramidal neurons and in thin, dense clusters of punctate profiles. In tangential sections through layer IVA, the three proteins are seen in distinct compartments. Calbindin- and parvalbumin immunostained neurons make up a thinly walled honeycomb or lattice, whereas neurons immunostained for nonphosphorylated neurofilament protein occupy the central lacunae. Direct comparison shows that neurons immunostained for calbindin occupy regions in layer IVA complementary to those immunostained for nonphosphorylated neurofilament protein. These data demonstrate a basic similarity in the organization of layer IV in macaques and humans. Layer IVA specifically is organized into complementary and neurochemically distinct compartments, including what appears to be a geniculocortically innervated and parvicellular-driven lattice and the interstitial lacunae formed by the periodic, upward protrusion of magnocellular-dominated layer IVB neurons. PMID- 7499526 TI - Efferent projections of the paraventricular thalamic nucleus in the rat. AB - The paraventricular nucleus of the thalamus (PVT) receives input from all major components of the circadian timing system, including the suprachiasmatic nucleus (SCN), the intergeniculate leaflet and the retina. For a better understanding of the role of this nucleus in circadian timing, we examined the distribution of its efferent projections using the anterograde tracer Phaseolus vulgaris leucoagglutinin (PHA-L). The efferent projections of the PVT are loosely organized along its dorsal-ventral and anterior-posterior axes. The anterior PVT sends projections to the SCN; the dorsomedial and ventromedial hypothalamic nuclei; the lateral septum; the bed nucleus of the stria terminalis; the central and basomedial amygdaloid nuclei; the anterior olfactory nucleus; the olfactory tubercle; the nucleus accumbens; the infralimbic, piriform, and perirhinal cortices; the ventral subiculum; and the endopiriform nucleus. A small PHA-L injection, restricted to the ventral portion of the anterior PVT, produces a similar pattern of labeling, except for a marked decrease in the number of labeled fibers in the hypothalamus, cortex, and lateral septum and an increase in labeling in the endopiriform nucleus and basolateral amygdaloid nucleus. The posterior PVT has a more limited efferent distribution than the anterior PVT, terminating in the anterior olfactory nucleus; the olfactory tubercle; the nucleus accumbens; and the central, basolateral, and basomedial nuclei of the amygdala. Our results show that the anterior PVT is ideally situated to relay circadian timing information from the SCN to brain areas involved in visceral and motivational aspects of behavior and to provide feedback regulation of the SCN. PMID- 7499527 TI - AMPA-selective glutamate receptor subtype immunoreactivity in the aged human hippocampal formation. AB - It has been hypothesized that, in Alzheimer's disease, glutamate-mediated excitotoxicity contributes to the degeneration of selected populations of neurons. In the present study, immunocytochemical techniques were used to determine the distribution and anatomical features of GluR1- and GluR2/3 immunolabeled cell bodies and processes within the hippocampal formation of normal (i.e., no pathology) elderly humans. The results of this study provide an essential baseline with which to compare the expression and distribution of glutamate receptor subunits within the brains of patients with Alzheimer's disease. With respect to GluR1 immunoreactivity, the molecular layer of the dentate gyrus displays the most intense immunolabeling of any hippocampal structure. Contributing to this intense labeling are apical dendrites that arise from neurons within the adjacent granule cell layer. Interestingly, GluR1-labeled neurons account for a relatively small percentage of the total number of neurons as revealed by Nissl staining in the granule cell layer. In contrast, GluR2/3 labeled neurons are densely distributed throughout the granule cell layer, yet they provide relatively few processes to the adjacent molecular layer compared to GluR1-positive processes. GluR1 labeling is also prominent within the CA fields of Ammon's horn, with CA2 > CA3 > CA1 > or = CA4. Most prominent within the CA fields are the labeled dendrites of pyramidal neurons. In many instances, apical dendrites can be traced into the adjacent stratum radiatum, where they impart a deep striated appearance to this region of the hippocampus. Robust GluR2/3 labeling is also observed within the pyramidal layer of Ammon's horn, with an order of staining intensity similar to that observed for GluR1. However, unlike GluR1 labeling, which is localized predominantly along dendrites, GluR2/3 labeling is observed primarily in association with cell bodies. Collectively, these data suggest that the molecular composition of the AMPA receptor complex may differ between the dendrite and soma of granule and pyramidal neurons within the hippocampal formation, so functionally we may predict that these two regions of the neuron would respond differently following glutamate receptor stimulation. PMID- 7499528 TI - Presynaptic metabotropic glutamate receptors in adult and developing neurons: autoexcitation in the olfactory bulb. AB - The mitral cell of the olfactory bulb is the primary relay neuron that transmits information from the olfactory receptors to the rest of the brain. This excitatory neuron releases glutamate from presynaptic dendrites and axon terminals. All rat mitral cells studied showed strong, selective, and widespread metabotropic glutamate receptor mGluR1 alpha immunoreactivity on the presynaptic membrane of dendrites, often at the synaptic vesicle release site, when examined with light and electron microscopy. The finding of glutamate receptors on mitral cell secondary dendrites supports the conclusion that not all dendritic membrane with glutamate receptors necessarily have gray type I asymmetrical synaptic specializations. In contrast, the metabotropic glutamate receptor mGluR5 was not found in mitral cells but was expressed by granule cells and astrocytes around mitral dendrites. Both mGluR1 alpha and mGluR5 were expressed early in development, with strong immunostaining present by postnatal day 1. MGluR1 alpha staining at birth mirrored the adult staining pattern. MGluR5 staining at birth showed different patterns of immunostaining than that found in the adult, particularly in the external plexiform layer. In vitro olfactory bulb neurons and their dendrites from embryonic day (E) 18 olfactory bulbs responded to t-ACPD and quisqualate, selective and nonselective metabotropic glutamate receptor agonists, and to several ionotropic glutamate agonists with increases in intracellular Ca2+ as studied with fura-2 digital imaging. These data indicate that the receptors were functionally active at an early stage of development. Application of the glutamate receptor blockers d-2-amino-5-phosphonovalerate (AP5) and 6-cyano-7 nitroquinoxaline (CNQX) to E17 olfactory bulb neurons after only 4 days in vitro resulted in a dramatic decrease in Ca2+ levels in 70% of 128 cells tested, suggesting that embryonic neurons after a short time in vitro can actively secrete glutamate. The presence of glutamate receptors on the long mitral cell dendrite suggests that it would be able to respond to release of its own excitatory transmitter, probably at an early stage of development. In the probable absence of other excitatory input to the secondary mitral dendrites, it would be the only excitatory "input." This autoexcitatory response would be modulated by release of GABA from olfactory interneurons occurring milliseconds after glutamate release induced by olfactory nerve activation. This novel type of neuronal microcircuitry would potentially amplify signal transmission and current spread along the long mitral dendrites and could play an important role in lateral inhibition of olfactory neurons. PMID- 7499529 TI - Visual areas in the dorsal and medial extrastriate cortices of the marmoset. AB - To define the number and limits of the visual areas in the primate extrastriate cortex, the visuotopy of the dorsal convexity and medial wall was studied by electrophysiological recordings in five marmosets anaesthetised with sufentanil and nitrous oxide and paralysed with pancuronium bromide. We identified five visuotopic representations in and around the densely myelinated zone between visual area 2 (V2) and the posterior parietal cortex. Most of the densely myelinated zone is formed by the homologue of the owl monkey's dorsomedial area (DM); thus, we also termed this area DM in the marmoset. Within DM, the lower quadrant representation is continuous, with central vision represented laterally, peripheral vision medially, the horizontal meridian caudally, and the vertical meridian rostrally. In contrast, the upper quadrant representation is split, with the central portion represented at the lateral edge of DM on the dorsal surface, and the periphery along the midline. Two other visual field representations, corresponding to the dorsointermediate area (DI) and to a new subdivision termed the dorsoanterior area (DA), are also densely myelinated but can be distinguished from DM based on the separation of the bands of Baillarger and visual topography. In addition, a homologue of the medial visual area (M) was identified. Our results reveal a highly complex visuotopy in primate cortex, with local discontinuities in representation and borders between areas that are often not coincident with either the horizontal or the vertical meridian. The topography of the dorsal extrastriate cortex in the marmoset strongly suggests that both visual area 3 (V3) and the parietooccipital area (PO) of other primates are portions of a single visuotopic representation, DM, and calls into question the existence of visual areas with partial or quadrantic representations of the visual field. PMID- 7499530 TI - Ultrastructural evidence for a paucity of projections from the lumbosacral cord to the pontine micturition center or M-region in the cat: a new concept for the organization of the micturition reflex with the periaqueductal gray as central relay. AB - Information concerning the rate of bladder filling is determined by receptors in the bladder wall and conveyed via afferent fibers in the pelvic nerve to sensory neurons in the lumbosacral cord. It was assumed that this information is relayed from the lumbosacral cord to a medial cell group in the dorsolateral pontine tegmentum, called the M-region, the pontine micturition center, or Barrington's nucleus. The M-region, in turn, projects via long descending pathways to the sacral parasympathetic motoneurons. In the present electron microscopic study, it was investigated in cats whether monosynaptic projections from lumbosacral neurons to the M-region indeed exist. Wheat-germ agglutinin-horseradish peroxidase injections were made into the lumbosacral cord. Many retrogradely labeled dendrites and somata were found in the M-region, but no labeled terminals were found on retrogradely labeled dendrites or somata. Only a small number of anterogradely labeled terminals, which were filled with mainly round vesicles, contacted unlabeled dendrites in the M-region. In contrast, many more anterogradely labeled terminals, which were filled with mainly round and, to a limited extent, dense core vesicles and with asymmetrical synapses, were found on dendrites in the lateral part of the periaqueductal gray (PAG). Previously (Blok and Holstege [1994] Neurosci. Lett. 166:93-96), it was demonstrated that the lateral part of the PAG contains neurons projecting to the M-region. A concept for the central organization of the micturition reflex is presented in which ascending projections from the lumbosacral cord convey information on bladder filling to the PAG.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7499531 TI - Thyrotropin-releasing hormone-immunoreactive varicosities synapse on rat phrenic motoneurons. AB - The relationship between retrogradely labelled or intracellularly filled phrenic motoneurons and varicosities containing thyrotropin-releasing hormone immunoreactivity was investigated in rats by light and electron microscopy. Phrenic motoneurons were identified via retrograde tracing from the diaphragm with cholera toxin B subunit, which was followed by immunocytochemistry to visualise retrogradely labelled motoneurons and thyrotropin-releasing hormone immunoreactive nerve fibres in their vicinity. At the light microscopic level, varicose thyrotropin-releasing hormone-immunoreactive nerve fibres were distributed sparsely in the phrenic motor nucleus, with some axons surrounding retrogradely labelled motoneurons. In separate intracellular experiments, four phrenic motoneurons identified by antidromic activation from the C5 phrenic nerve root were subsequently filled with Neurobiotin, and nerve fibres that contained thyrotropin-releasing hormone immunoreactivity were identified by immunocytochemistry. The numbers and locations of thyrotropin-releasing hormone immunoreactive varicosities that were closely appeared to the intracellularly labelled motoneurons were mapped using a camera lucida technique. Close appositions by thyrotropin-releasing hormone-immunoreactive varicosities were seen on somata as well as on proximal and distal dendrites. The closely apposed varicosities were usually present in tight clusters, which were formed by single varicose axons. However, the distribution was nonuniform, in that some dendrites did not receive any close appositions. Ultrastructural analysis of random ultrathin sections through retrogradely labelled neurons showed that varicosities with thyrotropin-releasing hormone immunoreactivity made 1.8% of all synapses and direct contacts on somata and 2.3% of synapses and contacts with dendrites of the retrogradely labelled phrenic motoneurons. The results of these experiments suggest that thyrotropin-releasing hormone-immunoreactive varicosities provide similar numbers of inputs to both the somata and dendrites of phrenic motoneurons. These thyrotropin-releasing hormone-containing inputs seen via light and electron microscopy could modulate the excitability of phrenic motoneurons. PMID- 7499532 TI - Distribution of acidic fibroblast growth factor mRNA-expressing neurons in the adult mouse central nervous system. AB - The distribution of acidic fibroblast growth factor (aFGF) mRNA-expressing neurons was studied throughout the adult mouse central nervous system (CNS) with in situ hybridization histochemistry using a radiolabelled synthetic oligodeoxynucleotide probe complementary to the mRNA of human aFGF. We report here a widespread distribution of aFGF mRNA in several defined functional systems of the adult mouse brain, whereby the highest levels of aFGF mRNA were found in large somatomotor neurons in the nuclei of the oculomotor, trochlear, abducens, and hypoglossal nerves; in the motoneurons of the ventral spinal cord and the special visceromotor neurons in the motor nucleus of the trigeminal nerve; and in the facial and ambiguus nuclei. Labelled perikarya were also detected in all central structures of the auditory pathway including the level of the inferior colliculus, i.e., the lateral and medial superior nuclei; the trapezoid, cochlear, and lateral lemniscal nuclei; and parts of the anterior colliculus. Furthermore, many aFGF-positive cell bodies were found in the vestibular system and other structures projecting to the cerebellum, in the deep cerebellar nuclei, in somatosensory structures of the medulla (i.e., in the gracile, cuneate, and external cuneate nuclei), as well as in the spinal nucleus of the trigeminal nerve. The findings that aFGF mRNA is expressed in all components of several well defined systems (i.e., in sensory structures) as well as in central neurons that process sensory information and, finally, in some efferent projections point towards a concept of aFGF expression primarily within certain neuronal circuitries. PMID- 7499533 TI - Dopamine transporter immunoreactivity in rat brain. AB - The dopamine transporter (DAT) is a primary site for the action of cocaine in inducing euphoria. Its action is necessary for the selectivities of dopaminergic neurotoxins that provide the best current experimental models of Parkinson's disease. In the present report, rat dopamine transporter-like immunoreactivity (iDAT) was assessed by immunohistochemistry using newly developed polyclonal antisera raised against conjugated peptides corresponding to sequences found in the dopamine transporter's carboxy- and amino-termini. Dense iDAT was observed in patterns consistent with neural processes and terminals in the striatum, nucleus accumbens, olfactory tubercle, nigrostriatal bundle, and lateral habenula. Perikarya in the substantia nigra pars compacta were immunostained with moderate intensity using one of two immunohistochemical methods, while scattered ventral tegmental area perikarya were stained with somewhat less intensity. Immunoreactive neuronal processes with axonal and dendritic morphologies were stained in the substantia nigra and the paranigral and parabrachialis pigmentosus nuclei of the ventral tegmental area, while sparser processes were noted more medially in the ventral tegmental area. Neuronal processes were found in several laminae in the cingulate cortex, with notable fiber densities in the superficial aspects of lamina I and laminae II/III. The intensities of immunoreactivities in striatum and cerebral cortex were dramatically attenuated ipsilateral to nigrostriatal bundle 6-hydroxydopamine lesions. Specificity of immunostaining was supported by agreement of the results using sera directed against two distinct DAT segments, studies with preimmune and preadsorbed sera and studies of the extracted protein. These antisera identify and reveal details of the distribution of DAT immunoreactivity in rat brain and display variations in levels of DAT expression of likely functional significance. PMID- 7499534 TI - Histological identification of premotor neurons for horizontal saccades in monkey and man by parvalbumin immunostaining. AB - The premotor excitatory and inhibitory burst neurons are essential for horizontal saccades. In the monkey, excitatory burst neurons lie in the ipsilateral paramedian pontine reticular formation, and the inhibitory burst neurons lie more caudally in the contralateral nucleus paragigantocellularis dorsalis. For a neuropathological analysis of degenerative changes in saccadic disorders of patients, the histological identification of the burst neuron areas in man is important. Here, we show that this is possible with parvalbumin immunostaining as a histological marker. First, in monkeys, the premotor burst neurons were backlabeled by injections of wheat germ agglutinin-horseradish peroxidase or cholera toxin subunit B into the abducens nucleus or tetanus toxin fragment C into the lateral rectus muscle and shown by double labeling to contain parvalbumin. Then, human brainstem sections were immunoreacted for parvalbumin, and, by comparing the resulting staining pattern to that in the monkey, the homologous burst neuron areas were defined in man. In the monkey, excitatory burst neurons were confirmed to the nucleus reticularis pontis caudalis and did not extend farther rostrally into the nucleus reticularis pontis oralis. All retrogradely labeled cells in both burst neuron areas were parvalbumin positive, and approximately 70% of the parvalbumin-positive cells were retrogradely labeled. Both burst neuron areas were highlighted by their parvalbumin staining pattern and could be outlined in man as well. The putative excitatory burst neuron area in man is in the medial part of the nucleus reticularis pontis caudalis (extending 2.5 mm mediolaterally), immediately rostral (250 microns) to the omnipause neurons and extending 2.2 mm rostrally, and the putative inhibitory burst neuron area lies in the medial part of the paragigantocellular nucleus caudal to the abducens nucleus, extending 1.8 mm caudally. The location of the burst neuron areas, including the burst neurons themselves, via parvalbumin immunostaining will help in the analysis of clinical cases with slow saccades. PMID- 7499535 TI - Expression of glial fibrillary acidic protein and its relation to tract formation in embryonic zebrafish (Danio rerio). AB - To address possible roles of glial cells during axon outgrowth in the vertebrate central nervous system, we investigated the appearance and distribution of the glial-specific intermediate filament, glial fibrillary acidic protein (GFAP), during early embryogenesis of the zebrafish (Danio rerio). Immunopositive cells first appear at 15 hours, which is at the time of, or slightly before, the first axon outgrowth in the brain. Immunopositive processes are not initially present in a pattern that prefigures the location of the first tracts but rather are distributed widely as endfeet adjacent to the pia, overlying most of the surface of the brain with the exception of the dorsal and ventral midline. The first evidence for a specific association of immunopositive cells with the developing tracts is observed at 24 hours in the hindbrain, where immunopositive processes border axons in the medial longitudinal fasciculus. By 48 hours, immunopositive processes have disappeared from most of the subpial lamina and are found exclusively in association with tracts and commissures in three forms: endfeet, radially oriented processes, and tangentially oriented processes parallel to axons. This last form is particularly prominent in the transverse plane of the hindbrain, where they define the boundaries between rhombomeres. These results suggest that glial cells contribute to the development and organization of the central nervous system by supporting early axon outgrowth in the subpial lamina and by forming boundaries around tracts and between neuromeres. The results are discussed in relation to previous results on neuron-glia interactions and possible roles of glial cells in axonal guidance. PMID- 7499536 TI - Sexual and seasonal differences in the vomeronasal epithelium of the red-backed salamander (Plethodon cinereus). AB - Sexually dimorphic behaviors often are associated with sexually dimorphic neural structures. Perinatal hormonal levels influence structural sexual dimorphism, and seasonal structural changes also can be the result of seasonal hormonal fluctuations. We compared the volume of vomeronasal organs of male and female red backed salamanders (Plethodon cinereus) throughout the year. Odorants are delivered to vomeronasal receptors through nasolabial grooves when salamanders touch the bases of these grooves to objects (nose tapping). Males may locate and identify potential mates through nose tapping during the prolonged breeding season that lasts from October through May. We compared vomeronasal organ data through multiple regression by using total body size, sex, and season as variables that may influence vomeronasal organ volume. Gonads also were examined as an indicator of reproductive status. Total body size and sex significantly affect vomeronasal organ volume; as body size increases, so does vomeronasal organ volume, and males have significantly larger vomeronasal organs than females at all times of the year. During the summer, both males and females have larger vomeronasal organs than at any other time of the year. Summer also is a period of intense foraging and the initiation of a new cycle of gametogenesis. Previous studies of circulating hormone levels in amphibians have shown that the initial stages of gametogenesis correspond to a period of low estradiol and testosterone levels but high gonadotropin-releasing hormone levels. The functional significance of sexual and seasonal differences in the vomeronasal organs of P. cinereus may relate to the neurogenesis of specialized receptors for courtship and mating. PMID- 7499537 TI - The process of reinnervation in the dentate gyrus of adult rats: gene expression by neurons during the period of lesion-induced growth. AB - Neurons in the hippocampal dentate gyrus are extensively reinnervated following the destruction of their normal inputs from the ipsilateral entorhinal cortex (EC). The present study evaluates gene expression by dentate granule neurons and the neurons giving rise to the sprouting connections during the period of synapse growth. Adult male rats were prepared for in situ hybridization at 2, 4, 6, 8, 10, 12, 14, 20, and 30 days following unilateral EC lesions. Sections were hybridized using 35S-labeled cRNA probes for mRNAs that encode proteins thought to be important for neuronal structure and/or synapse function, including (1) mRNAs that are normally present in dendrites--the mRNAs for the high molecular weight microtubule-associated protein 2 (MAP2) and the alpha-subunit of calcium/calmodulin-dependent protein kinase II (CAMII kinase), (2) mRNAs that are upregulated in neurons that are regenerating their axons (T alpha 1 tubulin and F1/GAP43) and (3) mRNAs for proteins that are the principal constituents of neurofilaments and microtubules (the low molecular weight neurofilament protein NF68 and beta-tubulin). Although there were small changes in the levels of labeling for the mRNAs that are normally present in dendrites, there were no dramatic increases in the levels of any of the mRNAs either in dentate granule cells or in neurons giving rise to the reinnervating fibers at any postlesion interval. These results indicate that neurons in mature animals can substantially remodel their synaptic terminals and their dendrites in the absence of large scale changes in gene expression (at least as measured by steady-state mRNA levels at various time points). PMID- 7499538 TI - Afferent influences on brainstem auditory nuclei of the chicken: regulation of transcriptional activity following cochlea removal. AB - Neuronal survival in the cochlear nucleus of young animals is regulated by afferent activity. Removal or blockade of nerve VIII input results in the death of 20-40% of neurons in the cochlear nucleus, nucleus magnocellularis (NM), of the 10-14 days posthatch chick. Neuronal death in NM is preceded by complete failure of protein synthesis and degradation of ribosomes. In addition, there is a biphasic change in the immunoreactivity of ribosomes for a monoclonal antiribosomal RNA antibody, Y10B. Initially, the entire population of afferent deprived NM neurons loses Y10B immunoreactivity, but, after 6 or 12 hours of afferent deprivation, lack of Y10B immunoreactivity specifically marks dying NM neurons. Whether RNA synthesis is also altered in afferent-deprived NM neurons has not previously been studied. To determine whether RNA synthesis in NM neurons is regulated by loss of afferent activity, we injected chicks with 3H-uridine following unilateral cochlea removal and measured the incorporation of RNA precursor with tissue autoradiography. As early as 1 hour after cochlea removal, there was a significant decrease in 3H-uridine incorporation by afferent-deprived NM neurons. After longer periods of afferent deprivation (6 or 12 hours), the majority of dying NM neurons (marked by loss of Y10B immunoreactivity) fail to incorporate RNA precursor. Six or 12 hours following cochlea removal, the subpopulation of surviving NM neurons incorporates 3H-uridine at increased levels over those observed 1 or 3 hours after cochlea removal. These findings suggest that nuclear function is regulated by afferent synaptic activity and that failure of RNA synthesis occurs early in the cell death process. PMID- 7499539 TI - Laminar distribution and sources of catecholaminergic input to the optic tectum of the pigeon (Columbia livia). AB - A combined immunohistochemical and retrograde tracing approach was used to characterize the catecholaminergic innervation of the optic tectum (TeO), the major target of retinal projections in many avian species. Giemsa counterstaining was employed to determine precisely the laminar localization of immunoreactive fibers and presumptive terminals. The TeO of the pigeon is densely innervated by fibers immunoreactive for tyrosine hydroxylase (TH), which are most heavily distributed to the superficial layers of its dorsal and anterior portions. Within the dorsal-anterior tectum, TH-immunoreactive processes are particularly dense in retinorecipient layers 4 and 7 and in layer 5a. As in the mammalian superior colliculus, the bulk of the catecholaminergic innervation of the pigeon TeO reflects inputs, presumably noradrenergic, originating in the locus coeruleus and nucleus subcoeruleus. However, the catecholaminergic innervation of the pigeon TeO shows several features distinct from those reported for the mammalian superior colliculus. These include an input from a pretectal TH-positive cell group unknown in mammals and the presence of residual TH immunoreactivity after administration of the noradrenergic neurotoxin DSP-4. Moreover, the pattern of TH immunoreactive fibers in pigeon TeO indicates more laminar and regional specialization within this structure than has been reported for the catecholaminergic innervation of the superior colliculus in mammals. PMID- 7499540 TI - Central nervous system neurons infected by pseudorabies virus injected into the rat urinary bladder following unilateral transection of the pelvic nerve. AB - Following the transection of one pelvic nerve and both hypogastric nerves, the urinary bladder of male Sprague-Dawley rats was injected with pseudorabies virus (PRV; Bartha strain). The central stump of the transected pelvic nerve was labelled with fast blue (FB), and rats were maintained for 2, 2.5, and 3 days following viral infection. Tissue was processed with antisera against PRV and choline acetyltransferase (CAT). In the L6-S1 spinal cord, neurons in the ipsilateral intermediolateral area (IML) were labelled after 2 days. After 2.5 days, labelled neurons were also found in the dorsal gray commissure (DGC), the ipsilateral superficial dorsal horn, and the contralateral IML area. After 3 days, many labelled neurons appeared in the superficial dorsal horns and, bilaterally, in the L6-S1 dorsal root ganglia. In both IMLs, two groups of PRV labelled neurons were found: 1) CAT-positive preganglionic cells and 2) smaller, CAT-negative cells located slightly dorsal to the preganglionic neurons. No other doubly stained neurons were found in the spinal cord. Contralateral DRG neurons stained for either PRV or FB or both. Ipsilateral DRG neurons stained only for PRV. PRV-immunoreactive (IR) neurons appeared in the brainstem only after 3 days. These were located primarily in the pontine micturition centers (equal numbers), the ventral locus coeruleus, and the raphe and lateral reticular areas. PMID- 7499541 TI - Caudal medullary pathways to lumbosacral motoneuronal cell groups in the cat: evidence for direct projections possibly representing the final common pathway for lordosis. AB - The nucleus retroambiguus (NRA) projects to distinct brainstem and cervical and thoracic cord motoneuronal cell groups. The present paper describes NRA projections to distinct motoneuronal cell groups in the lumbar enlargement. Lumbosacral injections of wheat germ agglutinin-horseradish peroxidase (WGA-HRP) were made to localize and quantify the retrogradely labeled neurons in the caudal medullary lateral tegmentum. These injections were combined with spinal hemisections to distinguish between neurons having ipsi-or contralaterally descending axons. The NRA-lumbosacral fibers descend almost exclusively contralaterally, but neurons in areas surrounding the NRA project mainly ipsilaterally. In an anterograde tracing study, injections of WGA-HRP or tritiated leucine were made in the region of the NRA to determine the NRA targets in the lumbosarcral cord. Hemisections in C2 made it possible to distinguish between NRA projections and projections from neurons in the adjoining lateral tegmentum. The results show delicate NRA projections to distinct lumbosacral motoneuronal cell groups innervating specific hindlimb muscles (iliopsoas, adductors, and hamstrings) as well as axial muscles (medial longissimus and proximal tail muscles). The projection is bilateral, with a contralateral predominance. Ipsilaterally terminating fibers are derived from NRA neurons whose axons cross the midline at the level of the obex, descend through the contralateral spinal white matter, and recross at the level of termination. A conceptual description is presented in which the periaqueductal gray-NRA lumbosacral projections form the final common pathway for lordosis in the cat. PMID- 7499543 TI - Human cingulate cortex: surface features, flat maps, and cytoarchitecture. AB - The surface morphology and cytoarchitecture of human cingulate cortex was evaluated in the brains of 27 neurologically intact individuals. Variations in surface features included a single cingulate sulcus (CS) with or without segmentation or double parallel sulci with or without segmentation. The single CS was deeper (9.7 +/- 0.81 mm) than in cases with double parallel sulci (7.5 +/- 0.48 mm). There were dimples parallel to the CS in anterior cingulate cortex (ACC) and anastomoses between the CS and the superior CS. Flat maps of the medial cortical surface were made in a two-stage reconstruction process and used to plot areas. The ACC is agranular and has a prominent layer V. Areas 33 and 25 have poor laminar differentiation, and there are three parts of area 24: area 24a adjacent to area 33 and partially within the callosal sulcus has homogeneous layers II and III, area 24b on the gyral surface has the most prominent layer Va of any cingulate area and distinct layers IIIa-b and IIIc, and area 24c in the ventral bank of the CS has thin layers II-III and no differentiation of layer V. There are four caudal divisions of area 24. Areas 24a' and 24b' have a thinner layer Va and layer III is thicker and less dense than in areas 24a and 24b. Area 24c' is caudal to area 24c and has densely packed, large pyramids throughout layer V. Area 24c' g is caudal to area 24c' and has the largest layer Vb pyramidal neurons in cingulate cortex. Area 32 is a cingulofrontal transition cortex with large layer IIIc pyramidal neurons and a dysgranular layer IV. Area 32' is caudal to area 32 and has an indistinct layer IV, larger layer IIIc pyramids, and fewer neurons in layer Va. Posterior cingulate cortex has medial and lateral parts of area 29, a dysgranular area 30, and three divisions of area 23: area 23a has a thin layer IIIc and moderate-sized pyramids in layer Va, area 23b has large and prominent pyramids in layers IIIc and Va, and area 23c has the thinnest layers V and VI in cingulate cortex. Area 31 is the cinguloparietal transition area in the parasplenial lobules and has very large layer IIIc pyramids. Finally, variations in architecture between cases were assessed in neuron perikarya counts in area 23a. There was an age-related decrease in neuron density in layer IV (r = -0.63; ages 45-102), but not in other layers.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 7499542 TI - Effect of sensory deafferentation on immunoreactivity of GABAergic cells and on GABA receptors in the adult cat visual cortex. AB - To investigate the effects of sensory deafferentation on the cortical GABAergic circuitry in adult cats, glutamic acid decarboxylase (GAD) and gamma-aminobutyric acid (GABA) immunoreactivity and GABA receptor binding were studied in the visual cortex of normal cats and compared with cats that had received restricted binocular central lesions of the retina and had survived for 2 weeks postlesion in a normal visual environment. In the visual cortex of lesioned cats, two changes were observed in the number of GAD-immunoreactive elements in the regions affected by the retinal lesions: the number of GAD-positive puncta decreased, whereas that of GAD-immunoreactive somata increased. In contrast, no detectable changes were measured in the number of GABA-immunopositive somata or puncta. At the receptor level, we observed no differences in either the laminar distribution or the affinity of cortical GABAA and GABAB receptors labeled with [3H]-muscimol and [3H]-baclofen, respectively, in the lesioned versus normal cats. We present the hypothesis that sensory deafferentation in these adult cats (1) leads to a reduction of cortical GABAergic inhibition in the deafferented region, and (2) that this decreased inhibition may permit changes in efficiency of synapses and (3) that these changes may represent a first stage of events underlying the retinotopic reorganization preceeding the structural changes. PMID- 7499544 TI - Development of the chiasm of a marsupial, the quokka wallaby. AB - We have previously shown that the mature optic chiasm of a marsupial is divided morphologically into three regions, two lateral regions in which ipsilaterally projecting axons are confined and a central region containing only contralaterally projecting axons. By contrast, in the chiasms of eutherian (placental) mammals studied to date, there is no tripartite configuration. Ipsilaterally and contralaterally projecting axons from each eye are mixed in the caudal nerve and in each hemichiasm and encounter axons from the opposite eye near the midline of the chiasm. Here, we show that, unlike eutherians, marsupials have astrocytic processes in high concentrations in lateral regions of the nerve and rostral chiasm. Early in development, during the period when optic axons are growing through the chiasm, many intrachiasmatic cells are seen with densities five to eight times higher in lateral than in central chiasmatic regions. Such cells continue to be added to all chiasmatic regions; later in development, considerably more are added centrally, as the chiasm increases in volume. In the mature chiasm, cell densities are similar in all regions. By contrast to the marsupial, cell addition in the chiasm of a placental mammal, the ferret, is almost entirely restricted to later developmental stages, after axons have grown through the chiasm, and there are no obvious spatial variations in the distribution of cells during the period examined. During development, similar to the adult marsupial, ipsilaterally projecting axons do not approach the chiasmatic midline but remain confined laterally. We propose that the cells generated early and seen in high densities in the lateral chiasmatic regions of the marsupial may play a role in guiding retinal axons through this region of pathway selection. These data suggest that there is not a common pattern of developmental mechanisms that control the path of axons through the chiasm of different mammals. PMID- 7499545 TI - Effects of early periods of monocular deprivation and reverse lid suture on the development of Cat-301 immunoreactivity in the dorsal lateral geniculate nucleus (dLGN) of the cat. AB - During certain sensitive periods early in postnatal life, the anatomical and physiological development of the central visual pathways of cats and monkeys can be affected by the nature of an animal's early visual experience. In the last few years, studies have been started on some of the molecular and biochemical events that underlie the many functional changes induced by early selected visual deprivation in the visual cortex of kittens. In this respect, the monoclonal antibody Cat-301 provides a potentially powerful tool, because it recognizes in the cat dorsal lateral geniculate nucleus (dLGN) a proteoglycan associated with the surface of a particular class of cells, namely Y cells. In the dLGN, the Cat 301 proteoglycan appears late in postnatal development, and it expression has been shown to be experience dependent in both the dLGN and visual cortex (M. Sur, D. Frost, and S. Hockfield, 1988, J. Neurosci. 8:874-882; A. Guimaraes, S. Zaremba, and S. Hockfield, 1990, J. Neurosci. 10:3014-3024). We have explored further the experience-dependent nature of Cat-301 expression in the dLGN with a view to establishing a biochemical correlate of the many functional changes induced by early monocular deprivation and its reversal in the kitten visual system. In addition to demonstrating differences in Cat-301 expression between deprived and nondeprived laminae of the dLGNs of kittens monocularly deprived to only 4 or 5 weeks of age, the magnitude of the laminar difference was found to increase as the period of deprivation was extended. Moreover, monocularly deprived kittens that subsequently received long periods of reverse lid suture exhibited a reversal of the pattern of immunoreactivity, so that the greatest immunoreactivity occurred in laminae innervated by the initially deprived eye. However, possibly the most surprising and important finding was the extremely low levels of immunoreactivity observed in both A laminae of monocularly deprived animals that had received relatively short periods of reverse lid suture. These data suggest that Y cell development can be drastically altered depending on the time of initiation of the period of reverse lid suture and its duration. PMID- 7499546 TI - Developmental patterning of rod and cone photoreceptors in embryonic zebrafish. AB - Cone photoreceptors in the zebrafish retina are arranged in a crystalline lattice, with each spectral subtype at a specific position in the array; rod photoreceptors are inserted around the cones. Patterning events and developmental mechanisms that lead to the formation of the cone mosaic are not known. To begin investigating this issue, we examined the initial stages of opsin expression in zebrafish embryos by in situ hybridization with goldfish opsin cRNA probes to determine how and when the cone mosaic pattern arises. We found both differences and similarities in the spatiotemporal patterns of rod and cone development, which suggest the following: 1) Expression of opsin message (including rod opsin, blue and red cone opsins) was found in a ventral patch of retina located nasal to the choroid fissure. 2) The cone mosaic pattern was generated by a crystallization-like process initiated in the precocial ventral patch and secondarily in nasal retina, which then swept like a wave into dorsotemporal retina. 3) The remainder of the retina, suggesting that these precocial rods might differ from typical rods. 4) Developmental maturation of rods in zebrafish, as reflected by expression of opsin, may be accelerated compared to cones, which are thought to become postmitotic before rods. These data are consistent with a model in which lateral inductive interactions among differentiating photoreceptors lead to patterning of the array. PMID- 7499547 TI - Intratelencephalic projections of the visual wulst in pigeons (Columba livia). AB - The visual wulst is the telencephalic target of the thalamofugal visual pathway of birds, and thus the avian equivalent of the striate cortex of mammals. The anterograde tracer Phaseolus vulgaris leucoagglutinin was used to follow the intratelencephalic connections of the major constituents of the visual wulst in pigeons. In particular, efferent pathways from the granular layer (Intercalated nucleus of the hyperstriatum accessorium, IHA), supragranular layer (hyperstriatum accessorium, HA), and infragranular layers (hyperstriatum intercalatus superior and/or hyperstriatum dorsale, HIS/HD) were investigated. These efferent projections were confirmed by injections of the retrograde tracer cholera toxin subunit B into their terminal fields. When a deposit of the anterograde tracer was centered in IHA, which receives the visual thalamic input, efferent fibers were seen mainly dorsomedially to IHA. When a deposit of the anterograde tracer was centered in HA, efferent fibers were seen to extend mainly in three directions: 1) medially to the tractus septomesencephalicus, which sends projections to extratelencephalic visual nuclei: 2) ventrolaterally to the lateral portion of the neostriatum frontale, where there were also labeled cells after the retrograde tracer was injected in HA; and 3) ventromedially to the paleostriatal complex, which is the avian equivalent of the mammalian caudale, 5) neostriatum intermedium, 6) archistriatum intermedium, and 7) hyperstriatum laterale. Finally, HIS/HD have projections predominantly to HA and the dorsocaudal telencephalon (area corticoidea dorsolateralis and area parahippocampalis), as well as relatively minor projections to the areas which also receive projections from HA. No anterogradely labeled fibers were seen in the tractus septomesencephalicus following the tracer injections in HIS/HD. These results indicate that the visual information from the granular layer is distributed via the supragranular layer HA to multiple areas within the telencephalon, such as the neostriatum frontale and paleostriatal complex. In addition, HA is the source of an extratelencephalic projection via the tractus septomesencephalicus. Thus, the avian supragranular layer HA contains neurons which are the source of both intratelencephalic and extratelencephalic projections, whereas neurons of the mammalian cortex are segregated into two distinct layers, supragranular and infragranular layers, based on the targets of their projections. The findings are further discussed and compared to the mammalian striate cortex. PMID- 7499548 TI - Nerve growth factor prevents apoptotic cell death in injured central cholinergic neurons. AB - Experimental lesions have been widely used to induce neuronal degeneration and to test the ability to trophic molecules to prevent lesion-induced alterations, but these studies have not demonstrated unequivocally that afflicted neurons die as a result of these manipulations. The documentation of neuronal death in the above described models and the time when it occurs after injury are crucial for the interpretation of trophic effects. In the present study, we combined multiple approaches to investigate the nature of retrograde neuronal changes in cholinergic neurons of the medial septal nucleus (MSN) after complete, unilateral transection of the fimbria-fornix (F-F). Projections neurons of the MSN were prelabeled with the fluorescent tracer Fluoro-gold (FG) 1 week prior to lesion. By counting both FG-labeled and choline acetyltransferase (ChAT)-immunoreactive neurons in the MSN at multiple time points postaxotomy, we differentiated the phenotypic response to injury from the degenerative process and established a critical time between the third and fourth weeks postaxotomy, during which approximately 50% of fluorescent perikarya disappear. Working in the previous time window, we identified dying cells by electron microscopy (EM) and terminal transferase-mediated (TdT) deoxyuridine triphosphate (d-UTP)-biotin nick end labeling (TUNEL) and showed that MSN neurons die via apoptosis, beginning at 16 days postaxotomy. An additional group of animals was allowed to survive for 1 month (i.e., 10 days after cell death has been completed); during this period, animals were treated with intraventricular nerve growth factor (NGF). Quantitative analysis of surviving cholinergic perikarya showed that NGF prevented degeneration of the majority of neurons. In concert, the results of the present study establish that NGF does not merely protect the phenotype but also prevents cell death in lesioned central cholinergic neurons. PMID- 7499549 TI - NGF-dependent and NGF-independent recovery of sympathetic function after chemical sympathectomy with 6-hydroxydopamine. AB - To help clarify the distinction between the nerve growth factor (NGF)-dependent collateral sprouting of sympathetic nerves and their NGF-independent regeneration after crush, we used 6-hydroxydopamine (OHDA) to destroy the sympathetic terminals in adult rats; this leaves the axons damage-free. Would recovery occur by regeneration and/or collateral sprouting? A single 6-OHDA injection abolished the sympathetic pilomotor field revealed by electrical stimulation of a cutaneous nerve. Recovery began within 2 days, and by 20 days the field was reestablished. If the field was "isolated" by adjacent denervations at the time of 6-OHDA treatment, the recovering pilomotor field expanded extensively into the surrounding territory. In the presence of anti-NGF, however, the pilomotor field expansion ceased at about 60% of its former size; if anti-NGF treatment was discontinued, expansion recommended and extended into the surrounding skin. We suggest that the latter, NGF-dependent, growth phase corresponds to collateral sprouting, and the initial NGF-independent one to regeneration. After simple nerve crush, however, such regeneration can triple the normal sympathetic field size. This difference between crush- and 6-OHDA-induced regeneration might relate to the "cell body reaction" (CBR); the CBR is reduced with increasing distance of the lesion and is undetectable after a 6-OHDA lesion. Since the CBR and the vigor of regeneration are both increased by repeated axonal injury, we tested the effects of multiple 6-OHDA treatments; this significantly increased the initial NGF-independent expansion. We hypothesize that regeneration is regulated largely by mechanisms associated with the CBR, and that neurotrophin-dependent collateral sprouting occurs independently of these. PMID- 7499551 TI - Cells of the perireticular nucleus project to the developing neocortex of the rat. AB - The perireticular nucleus is a recently described thin sheet of small cells among the fibres of the internal capsule, lying lateral to the thalamic reticular nucleus and medial to the globus pallidus (Clemence and Mitrofanis [1992]. J. Comp. Neurol. 322:167-180). During development, the perireticular nucleus is relatively large, lying in the path of the growing corticofugal and thalamocortical axons and filling the area of the internal capsule lateral to the thalamic reticular nucleus. After these axons have formed their connections, the perireticular nucleus rapidly decreases in size, leaving only a few cells in the adult (Mitrofanis [1992] J. Comp. Neurol. 320:161-181). In this study, we aimed to investigate the connections between the developing cortex and thalamus by making injections of tracer into the cortical plate. Injections of Horse Radish Peroxidase (HRP), Wheat Germ Agglutinin bound to HRP (WGA-HRP) and 1'dioctadecyl 3,3,3',3 tetramethycarbocyanine perchlorate (DiI) were made in vivo between embryonic day (E) 18 and adult and DiI was placed in the fixed brains of rats aged between E16 and postnatal day (P)1. Between E17 and P10, the retrograde perikaryal labelling resulting from these injections revealed a transient projection from the perireticular nucleus to the ipsilateral cortical plate. No cells were labelled in the thalamic reticular nucleus. This suggests that the perireticular nucleus must be regarded as a group of cells distinct from the thalamic reticular nucleus and having a separate role in development. Comparisons between the perireticular cells and the cells of the cortical subplate suggest that both may be playing comparable roles in early development, possibly guiding fibres towards their end stations or serving to rearrange the complex mapped projections linking the thalamus and cortex. PMID- 7499552 TI - Distribution in the central nervous system of Aplysia of afferent fibers arising from cell bodies located in the periphery. AB - The present study using autoradiography to determine the location of the projections of presumptive peripheral afferent neurons into the central nervous system of Aplysia. Selected peripheral tissues (with an emphasis on structures involved in feeding behavior) were exposed to radioactive amino acids, and the distribution of macromolecules transported into the nervous system via afferent fibers was determined by autoradiography. Different regions of the body exhibited different patterns of projections, and, within the neuropil of the cerebral ganglion, there was a loose topographical organization of projections from the head. For some regions of the body, the projections was largely limited to the ganglion from which the nerve enters; for other regions, the projection was very widespread. In some cases (e.g., rhinophore to eye), there was evidence of projections from one peripheral structure to another. Experiments with all peripheral tissues that were studied resulted in extensive labeling of central ganglia, indicating that afferents with peripheral cell bodies may provide a major source of sensory input to the central nervous system and suggesting that many or all of the numerous ultrafine axons visualized via electron microscopy in the nerves of Aplysia may originate from first- or second-order sensory afferents whose cell bodies are located in the periphery. PMID- 7499553 TI - Macrophages and glia participate in the removal of apoptotic neurons from the Drosophila embryonic nervous system. AB - Cell death in the Drosophila embryonic central nervous system (CNS) proceeds by apoptosis, which is revealed ultrastructurally by nuclear condensation, shrinkage of cytoplasmic volume, and preservation of intracellular organelles. Apoptotic cells do not accumulate in the CNS but are continuously removed and engulfed by phagocytic haemocytes. To determine whether embryonic glia can function as phagocytes, we studied serial electronic microscopic sections of the Drosophila CNS. Apoptotic cells in the nervous system are engulfed by a variety of glia including midline glia, interface (or longitudinal tract) glia, and nerve root glia. However, the majority of apoptotic cells in the CNS are engulfed by subperineurial glia in a fashion similar to the microglia of the vertebrate CNS. A close proximity between macrophages and subperineurial glia suggests that glia may transfer apoptotic profiles to the macrophages. Embryos affected by the maternal-effect mutation Bicaudal-D have no macrophages. In the absence of macrophages, most apoptotic cells are retained at the outer surfaces of the CNS, and subperineurial glia contain an abundance of apoptotic cells. Some apoptotic cells are expelled from the CNS, which suggests that the removal of apoptotic cells can occur in the absence of macrophages. The number of subperineurial glia is unaffected by changes in the rate of neuronal apoptosis. PMID- 7499550 TI - Complementary expression of parvalbumin and calbindin D-28k delineates subdivisions of the rabbit medial geniculate body. AB - The complementary pattern of immunohistochemical staining for the calcium-binding proteins parvalbumin (PV) and calbindin D-28k (CB) was used to delineate four major subdivisions of the rabbit medial geniculate body (MGB). PV immunoreactivity predominates in the ventral and medial divisions, whereas CB immunoreactive cells characterize the dorsal and internal divisions. The ventral nucleus is strongly PV+ due to dense neuropil labeling and moderately labeled somata. The medial nucleus contains both medium-sized and large PV+ somata, as well as thick PV+ axons and terminals. The wedge-shaped internal nucleus composed of densely labeled CB+ cells, separates the dorsal and ventral nuclei rostrally, and expands caudally to encapsulate the posterior MGV. Large multipolar CB+ neurons with radiate dendrites characterize the dorsal nucleus. The differential expression of PV and CB also distinguishes the deep dorsal and superficial dorsal subnuclei in the dorsal division and a ventrolateral component in the ventral division. A comparison with studies of MGB connectivity in a variety of species suggests that PV immunoreactivity is highest in subdivisions that receive a substantial input from the central nucleus of the inferior colliculus and that project to primary auditory cortex. In contrast, CB immunoreactivity characterizes nuclei that receive input primarily from other sources, such as the paracentral nuclei of the inferior colliculus, the lateral tegmentum, and the spinal cord, and that project to secondary auditory areas. The ability of calcium binding protein immunohistochemistry to delineate neuronal compartments across indistinct cytoarchitectonic borders makes it a powerful tool for guiding future connectional and physiological studies of the MGB. PMID- 7499554 TI - Cell body response to injury in motoneurons and primary sensory neurons of a mutant mouse, Ola (Wld), in which Wallerian degeneration is delayed. AB - We examined the response to axon injury in the facial motoneurons and dorsal root ganglion (DRG) neurons of C57BL/Ola (Wld) mice, compared with the responses of C57BL/6J mice. The peripheral nerves of Ola mutants undergo remarkably slowed and muted Wallerian degeneration after injury. The increase in GAP-43 mRNA levels in facial motoneurons and DRG neurons was similar in both strains of mice, as was the initial decrease in medium-weight neurofilament (NFM) mRNA in facial motoneurons, and the increase in JUN immunoreactivity in both types of neurons. However, the subsequent recovery to normal low levels of JUN and GAP-43 mRNA expression and high levels of NFM mRNA was delayed in Ola motoneurons. We ascribe this delay to the slow regeneration and target reinnervation of facial axons in the Ola mice. These results show that absence of rapid Wallerian degeneration does not affect the initial cell body response to axonal injury. They also provide further evidence that restoration of normal levels of expression of GAP 43 and NFM mRNAs is dependent on target reinnervation and/or trophic factors provided by the distal nerve. Impaired regeneration in the Ola mouse does not seem to be a consequence of a defective cell body response to injury, and our results illustrate the general principle that, even if there is a vigorous cell body response to injury, normal axonal regeneration requires the additional provision of a favorable environment for growth. PMID- 7499555 TI - Differential regenerative response of Purkinje cell and inferior olivary axons confronted with embryonic grafts: environmental cues versus intrinsic neuronal determinants. AB - Regeneration of severed central axons is supposed to depend on two factors: a permissive local environment and the particular intrinsic properties of axotomized neurones. To assess the role of each of these factors in axonal regeneration, the capability of two particular axon populations of the adult mouse cerebellum to grow into target-specific (cerebellum) and target-unspecific (neocortex) embryonic grafts was determined. Purkinje cell and inferior olivary axons were transected by passing a microscalpel through the axial white matter of the cerebellar folia, particularly those of the anterior lobe. Immediately after the injury, solid transplants were placed in the lesion cavity. Purkinje cell axons were labelled by using anticalbindin immunocytochemistry, and olivocerebellar fibres were visualized by biotinylated dextran amine anterograde axonal tracing. Following axotomy, Purkinje cell axons appeared as thickened processes ending with large terminal clubs. Their morphology and number did not change up to the longest survival time considered (2 months), thereby confirming previous demonstrations that Purkinje cells survive axon injury (I. Dusart and C. Sotelo, 1994, J. Comp. Neurol. 347:211-232). Inferior olivary axons were thinner and bore smaller terminal bulbs. When embryonic cerebellar grafts, containing cortical and deep nuclear precursors, were placed close to the injured axons, olivocerebellar fibres vigorously regenerated into the transplants and ended in new climbing fibres along the dendrites of grafted Purkinje cells. By contrast, host Purkinje cell axons never showed any outgrowth towards the graft. Similarly, these axons failed to regenerate into grafts containing solely the rostromedial portion of the cerebellar anlage, mostly consisting of deep nuclear neurones, their main targets. Comparable results were obtained by transplanting embryonic neocortical tissue: inferior olivary axons also regenerated into the grafts, although with distinct terminal arbours without the climbing fibre phenotype, whereas Purkinje cell axons always failed to grow. These results provide the first direct demonstration that severed inferior olivary axons are able to regenerate. In addition, they show that the growth-permissive/-promoting conditions created by embryonic nervous tissue are not sufficient to induce the regeneration of every axonal type and allow us to hypothesise that successful regeneration depends on the interplay between environmental cues and intrinsic properties of the axotomized neurones. PMID- 7499556 TI - Vitronectin receptor expression and distribution at the photoreceptor-retinal pigment epithelial interface. AB - Laser scanning confocal microscopy was employed to map the distribution of integrin immunoreactivity at the photoreceptor-retinal pigment epithelial (RPE) interface of the primate retina, and to determine its relationship to the actin cytoskeleton. Immunolabeling using a polyclonal antibody to the human vitronectin receptor (VnR), a heterodimer containing the alpha v subunit in combination with either the beta 3 or beta 5 subunits, is detected primarily on the apical surface of the retinal pigment epithelium (RPE) in vivo and in vitro. It is also associated with the photoreceptor inner and outer segment cell surfaces. In contrast, immunolabeling using a polyclonal antibody to the human fibronectin receptor (FnR), a heterodimer containing the alpha 5 and beta 1 subunits, is detected principally on the basolateral surface of the RPE and is virtually absent in photoreceptors. A partial three-dimensional reconstruction of the anti VnR labeling pattern in cone photoreceptors reveals cell surface labeling that originates at the level of the myoid just distal to the outer limiting membrane. It extends distally toward the ellipsoid and terminates at the level of the cone outer segment. Approximately 20-22 immunoreactive foci are distributed evenly around the perimeter of the cone ellipsoid. These foci correspond in number and location to the calycal processes that protrude from the distal portion of the ellipsoid. A double-labeling procedure, employing VnR antibody and a fluorescently labeled phallotoxin (phalloidin), was used to identify regions of VnR co-distribution with filamentous actin (F-actin). One such region includes the VnR-immunoreactive foci at the margins of the cone inner segments and the actin cables that course through the photoreceptor ellipsoid and terminate within the calycal processes. A second zone of co-distribution coincides with the actin containing, circumferential bundle at the lateral borders of the RPE cells, and a third zone is associated with the apical microvilli of the RPE that ensheath cone outer segments. In order to help identify the specific subunits underlying VnR (alpha v beta 3/5) immunoreactivity, Northern blots of retinal-RPE RNA were probed with alpha 32P-cDNAs to the human alpha v, beta 3, and beta 5 subunits and additional immunolocalization studies were performed using integrin human alpha or beta subunit-specific antisera. The results from these studies strongly suggest that one or more integrins, containing the alpha v and/or beta 5 subunits, are expressed by the photoreceptors and RPE.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 7499557 TI - Time-course of neuropeptide changes in peri-ischemic zone and amygdala following focal ischemia in rats. AB - Previously, using a middle cerebral artery occlusion model in Wistar rat, we showed autonomic disturbances similar to those seen clinically and observed striking neurochemical changes in cortical and subcortical sites at 5 days following stroke. The neurochemical changes may account for functional recovery and/or autonomic disturbances after focal ischemia. To understand the possible mechanisms and to facilitate future studies, it is necessary to define the time courses of these changes. Using immunohistochemical staining with the peroxidase antiperoxidase reaction, the changes in several neuropeptides over the peri ischemic region and the ipsilateral central and basolateral nucleus of the amygdala were investigated at different times after middle cerebral artery occlusion. In the experimental group, neuropeptide Y immunoreactivity appeared to increase by 6 hours in the peri-ischemic region. Using image analysis to quantify the staining intensity, the change became statistically significant at 1 day, peaked around 3 days, and subsided at 10 days. There was a delayed increase in neuropeptide Y in the ipsilateral basolateral nucleus of the amygdala with a peak around 3 days. Immunoreactive staining for leucine-enkephalin, dynorphin, and neurotensin demonstrated an increase that was localized to the ipsilateral central nucleus of the amygdala with a peak around 3 days and a return to baseline levels by 10 days. The results support a specific time-course for each of the neuropeptides studied and indicate that a survival time of 3 days after focal ischemia is the critical period for examining the relationship between neuropeptide responses and neuronal or functional recovery. PMID- 7499558 TI - Reorganization of central terminals of myelinated primary afferents in the rat dorsal horn following peripheral axotomy. AB - We have investigated the time course and extent to which peripheral nerve lesions cause a morphological reorganization of the central terminals of choleragenoid horseradish peroxidase (B-HRP)-labelled primary afferent fibers in the mammalian dorsal horn. Choleragenoid-horseradish peroxidase is retrogradely transported by myelinated (A) sensory axons to laminae I, III, IV and V of the normal dorsal horn of the spinal cord, leaving lamina II unlabelled. We previously showed that peripheral axotomy results in the sprouting of numerous B-HRP-labelled large myelinated sensory axons into lamina II. We show here that this spread of B-HRP labelled axons into lamina II is detectable at 1 week, maximal by 2 weeks and persists for over 6 months postlesion. By 9 months, however, B-HRP fibers no longer appear in lamina II. The sprouting into lamina II occurs whether regeneration is allowed (crush) or prevented (section with ligation), and does not reverse at times when peripheral fibers reinnervate the periphery. We also show that 15 times more synaptic terminals in lamina II are labelled by B-HRP 2 weeks after axotomy than in the normal. We interpret this as indicating that the sprouting fibers are making synaptic contacts with postsynaptic targets. This implies that A-fiber terminal reorganization is a prominent and long-lasting but not permanent feature of peripheral axotomy. We also provide evidence that this sprouting is the consequence of a combination of an atrophic loss of central synaptic terminals and the conditioning of the sensory neurons by peripheral axotomy. The sprouting of large sensory fibers into the spinal territory where postsynaptic targets usually receive only small afferent fiber input may bear on the intractable touch-evoked pain that can follow nerve injury. PMID- 7499559 TI - Projections from the cochlear nucleus to the superior paraolivary nucleus in guinea pigs. AB - Axonal tracing techniques were used to study the projection from the cochlear nucleus to the superior paraolivary nucleus in guinea pigs. Different tracers were used to identify the cell types that give rise to the projections, the morphology of their axons, and the cell types that they contact in the superior paraolivary nucleus. Injections of Fluoro-Gold or peroxidase-labeled-WGA and HRP into the superior paraolivary nucleus labeled multipolar cells and octopus cells bilaterally in the ventral cochlear nucleus, mainly on the contralateral side. Injections of PHAL into the ventral cochlear nucleus labeled two types of axons in the superior paraolivary nucleus. Thin axons branch infrequently and give rise primarily to small, en passant boutons. Thick axons have larger boutons, many of which are terminal boutons that arise from short collaterals. Thin axons appear to originate from multipolar cells, whereas thick axons probably originate from octopus cells. Both types are found bilaterally after an injection into the ventral cochlear nucleus on one side. Individual thick or thin axons may contact multiple cell types in the superior paraolivary nucleus. Individual cells in the superior paraolivary nucleus can receive convergent input from both thick and thin axons. Combined anterograde and retrograde transport of different fluorescent tracers was used to identify the projections of the cells in the superior paraolivary nucleus that receive inputs from the ventral cochlear nucleus. Cells in the superior paraolivary nucleus that projected to the ipsilateral cochlear nucleus or to the ipsilateral inferior colliculus appeared to be contacted by axons that were labeled by anterograde transport from the contralateral ventral cochlear nucleus. Thus the projections to the superior paraolivary nucleus are in a position to affect the activity in both ascending and descending auditory pathways. PMID- 7499560 TI - Dendritic morphology of CA1 pyramidal neurones from the rat hippocampus: I. Branching patterns. AB - The aim of this study was to provide quantitative descriptions of the dendritic branching patterns of pyramidal neurones in the CA1 region of the rat hippocampus. Thirteen adult cells were filled with biocytin and reconstructed by using the light microscope. The number of basal trees arising from the soma of each cell ranged from two to eight. There was wide variation in the number of terminal segments per tree. Six cells had single apical trunks, and seven had trunks that bifurcated in stratum radiatum. The number of apical oblique trees ranged from nine to 30, with each tree usually showing a lower degree of branching than basal trees. Basal and oblique trees had similar branching patterns, with the majority of branch points occurring close to the origin of the tree. Both basal and oblique terminal segments were generally much longer than intermediate segments and constituted up to 90% of the combined dendritic length of the tree. The branching pattern of the apical tuft was different, with many relatively long intermediate segments; terminal segments contributed only some 66% of the combined dendritic length of these trees. The mean total combined dendritic length for six adult cells reconstructed and measured completely was 11,900 +/- 1,000 microns (standard deviation). The relative proportions of the different parts of the dendritic system, although not the total dendritic length, were correlated with the location of the soma relative to the cell body layer. Cells with somata close to the stratum pyramidale/stratum radiatum border had more dendrites terminating in stratum radiatum and fewer in stratum oriens than cells with somata further from it. PMID- 7499561 TI - Dendritic morphology of CA1 pyramidal neurones from the rat hippocampus: II. Spine distributions. AB - The numbers and distributions of dendritic spines were estimated for six adult and three juvenile biocytin-injected neurones from the CA1 region of the hippocampus of the albino rat. For each cell, a sample of long dendritic segments that lay favourably in the plane of focus was drawn at high magnification and the visible spines counted. Correction was made for spines obscured by dendritic shafts. Within individual cells, dendrites of similar type and diameter had similar spine densities. For adults, long basal segments averaged 2.4 spines/microns and obliques averaged 3.2 spines/microns. In juveniles, basals averaged 2.3 spines/microns and obliques, 2.5 spines/microns. Apical tuft segments were less spiny, averaging 1.4 spines/microns in adult cells and 1.8 spines/microns in juveniles. There was a positive correlation between spine density and dendrite diameter. Values from this sample were used to assign spine densities to the other segments, and so the total number of spines was estimated for each cell. Adult cells averaged 30,500 +/- 3,900 (S.D.) spines and juveniles, 23,800 +/- 7,100 spines. Adult cells had roughly 50% of their spines in stratum radiatum, 40% in s. oriens, and 10% in s. lacunosum-moleculare. Juvenile cells had a rather higher proportion (20%) in s. lacunosum-moleculare. In general, some 50% of all spines were located within a path length of 200 microns from the soma. These total numbers of spines were much higher than earlier values from Golgi impregnated cells but align well with estimates of the numbers of axonal boutons supplied to CA1 by CA3 pyramidal cells. PMID- 7499562 TI - Axonal projections from the lateral and medial superior olive to the inferior colliculus of the cat: a study using electron microscopic autoradiography. AB - The superior olivary complex is the first site in the central auditory system where binaural interactions occur. The output of these nuclei is direct to the central nucleus of the inferior colliculus, where binaural inputs synapse with monaural afferents such as those from the cochlear nuclei. Despite the importance of the olivary pathways for binaural information processing, little is known about their synaptic organization in the colliculus. The present study investigates the structure of the projections from the lateral and medial superior olivary nuclei to the inferior colliculus at the electron microscopic level. Stereotaxic placement and electrophysiological responses to binaural sounds were used to locate the superior olive. Anterograde axonal transport of 3H leucine was combined with light and electron microscopic autoradiography to reveal the location and morphology of the olivary axonal endings. The results show that the superior olivary complex contributes different patterns of synaptic input to the central nucleus of the inferior colliculus. Each projection from the superior olivary complex to the colliculus differs in the number and combinations of endings. Axonal endings from the ipsilateral medial superior olive were exclusively the round (R) type that contain round synaptic vesicles and make asymmetrical synaptic junctions. This morphology is usually associated with excitatory synapses and neurotransmitters such as glutamate. Endings from medial superior olive terminate densely in the central nucleus. The projection from the contralateral lateral superior olive also terminates primarily as R endings. This projection also includes small numbers of pleomorphic (PL) endings that contain pleomorphic synaptic vesicles and usually make symmetrical synaptic junctions. The PL morphology is associated with inhibitory synapses and transmitters such as gamma-aminobutyric acid and glycine. All endings from the contralateral lateral superior olive terminate much less densely than endings from the medial olive. In contrast, the projection from the ipsilateral lateral superior olive contributes both R and PL endings in roughly equal proportions. These ipsilateral afferents are heterogeneous in density and can terminate in lower or higher concentrations than endings from the contralateral side. These data show that the superior olive is a major contributor to the synaptic organization of the central nucleus of the inferior colliculus. The ipsilateral projections of the medial and lateral superior olive may produce higher concentrations of R endings than other inputs to the central nucleus. Such endings may participate in excitatory synapses. The highest concentrations of PL endings come from the ipsilateral lateral superior olive.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 7499563 TI - An atlas of aromatase mRNA expression in the zebra finch brain. AB - Neural conversion of androgen to estrogen by aromatase is an important step in the development and expression of masculine behavior in mammals and birds. In contrast to the low telencephalic levels of aromatase in adult mammals and nonsongbirds, the zebra finch telencephalon possesses high aromatase activity. This study maps, by in situ hybridization, cells that express aromatase mRNA in the adult zebra finch telencephalon, diencephalon, midbrain, and pons. High aromatase mRNA expression was observed in the caudal neostriatum, limbic archistriatum, and hypothalamus. The hippocampus, parahippocampal area, and hyperstriatum accessorium contained cells expressing moderate amounts of aromatase message. Weakly labeled cells were found in the rostral neostriatum, lobus parolfactorius, and mesencephalic reticular formation. These findings are consistent with aromatase activity measurements of zebra finch tissue and document with anatomical precision both the widespread expression of aromatase mRNA in the brain and novel sites of brain aromatase. This study identifies the caudal neostriatum as a major site of telencephalic aromatase. A previous survey (Gahr et al., 1993: J. Comp. Neurol. 327:112-122) of several avian species found that the presence of estrogen receptors in parts of the caudal neostriatum is unique to songbirds, which are the only birds to possess the elaborated telencephalic song system. Together, these findings suggest that the heightened estrogen synthesis and estrogen sensitivity of the passerine caudal neostriatum may have some functional relation with the telencephalic circuits responsible for song. PMID- 7499565 TI - 5'-nucleotidase in the rodent ventral striatum: relation to the distribution of leu-enkephalin, cell clusters, and infralimbic cortical innervation. AB - The goal of this study was to determine the compartmental organization of 5' nucleotidase within the rodent ventral striatum and to compare the distribution of 5'-nucleotidase with that of leu-enkephalin, cell clusters, and infralimbic cortical innervation. In the core, 5'-nucleotidase is present in several contiguous patchy structures that are in register with leu-enkephalin compartments. In the shell, 5'-nucleotidase is concentrated in a longitudinal band along the septal border. This "medial band" extends from the rostral pole of the ventral striatum to the bed nucleus of stria terminalis. The ventral portion of the medial band is in register with a cluster of cells, located medial to the most dorsal island of Cajella. A second 5'-nucleotidase compartment along the border of the core and shell is in register with a cell cluster and is most evident at caudal levels of the ventral striatum. The innervation of the ventral striatum by the infralimbic cortex is denser in the shell than in the core. In the shell, fibers from the superficial layers of the infralimbic cortex tend to avoid the 5'-nucleotidase-rich cell clusters and terminate in areas of moderate 5'-nucleotidase density. By contrast, fibers from the deep layers terminate in the ventral striatum without regard to the 5'-nucleotidase-rich cell clusters. Overall, the compartmental structure of 5'-nucleotidase in the ventral striatum segregates projections from different layers of the infralimbic cortex. Dense 5' nucleotidase compartments are innervated by neurons in the deep layers of the infralimbic cortex. The area of moderate 5'-nucleotidase density surrounding the 5'-nucleotidase compartments is innervated by neurons in both the superficial and deep layers. PMID- 7499564 TI - Development of catecholamine systems in the central nervous system of the newt Pleurodeles waltlii as revealed by tyrosine hydroxylase immunohistochemistry. AB - The aim of the present study was to extend our knowledge of the development of catecholamine (CA) systems in the class of amphibians to the order of urodeles. In contrast to previous studies of urodeles, the present study with antisera against tyrosine hydroxylase (TH) and dopamine revealed that CA systems are already present at early embryonic stages of the newt, Pleurodeles waltlii. Although the development from fertilized egg to juvenile in the urodele Pleurodeles lasts twice as long as that in the anuran, Xenopus laevis, and shows less dramatic changes in external morphology, the spatiotemporal sequence of appearance of TH-immunoreactive cell groups is rather similar. An early appearance of TH-immunoreactive cell bodies occurs in the olfactory bulb, the posterior tubercle, the accompanying cell group of the hypothalamic periventricular organ, the suprachiasmatic nucleus, the locus coeruleus, an area immediately ventral to the central canal of the spinal cord, and in the retina. Somewhat later, immunoreactive cells are detected in the posterior thalamic nucleus and in the rostral portion of the midbrain tegmentum, whereas the preoptic cell group is the last one to become TH immunoreactive. The presence of CA systems at early embryonic stages of both anurans and urodeles suggests that these systems are already of functional significance early in development. The maturation of CA neuronal structures in the olfactory and retinal circuitries, which takes place during development earlier in amphibians than in mammals, supports that notion. PMID- 7499566 TI - Platelet-derived growth factor receptors of mouse central nervous system cells in vitro. AB - This study evaluates the distribution of receptors for platelet-derived growth factor (PDGF) on central nervous cells maintained in vitro using colloidal gold labeled immunocytochemical markers at the electron microscopic level. Platelet derived growth factor receptors were found to be sparsely distributed over the surface of type 1 astrocytes, apparent type 2 astrocytes, and neurons. Receptors appeared to be preferentially associated with filopodia-like extensions of the cell membrane. The existence of functional receptors was confirmed using the impermeant, water-soluble affinity cross-linking agent bis(sulfosuccinimidyl)suberate to covalently link radiolabeled PDGF to its receptor. The PDGF/receptor complexes could also be immunoprecipitated with the same antibody used in immunocytochemical experiments. The improved resolution of these techniques allows definitive identification of PDGF receptors on cultured mammalian central nervous system cells other than oligodendrocytes. These data expand the range of possible roles of PDGF during nervous system development. Receptors for PDGF are likely to play a key role in the differentiation of cells in the central nervous system. PMID- 7499567 TI - Electroreceptor development in the electric fish Eigenmannia: a histological and ultrastructural study. AB - Developmental patterns of electroreceptors in the weakly electric fish Eigenmannia were investigated by histological, histochemical, immunocytochemical, cell kinetic, ultrastructural, and computer-assisted three-dimensional reconstruction methods. The first cell of an electroreceptor primordium is embedded in the stratum germinativum of the epidermis. An unmyelinated, afferent nerve fiber ends near this cell below the basal lamina. Protrusions and vacuole like inclusions at the basal lamina above the nerve fiber ending suggest a mechanism of nervous induction. The receptor primordium cell subsequently divides into a single cell layer. Within 48 hours, a second apical cell layer forms from the first, and, thus, the primordium differentiates into an apical layer of presumptive receptor cells and a basal layer of presumptive supporting cells. While the two layers further differentiate into mature receptor and supporting cells, the afferent fiber penetrates the basal lamina, sprouts, and forms a synapse with each receptor cell. Transitory fibers also project along the receptor cells to the top of the developing electroreceptor but degenerate during development. Synapses are smaller in early developmental stages compared to older stages, and pre- and postsynaptic vesicles are more abundant and widely distributed in younger stages. Moreover, presynaptic ribbons are longer and are interconnected at their apical ends. Supporting cells continue to divide during further maturation and form new receptor cells. The number of receptor cells per tuberous organ increases during the first 4 days of electroreceptor development and plateaus when the fish are 9 days old. It declines again when organs begin to divide into clusters. PMID- 7499568 TI - Localized reactions to injected therapeutic materials. Part 2. Surgical agents. PMID- 7499569 TI - Comparison of p53 expression in dermatofibrosarcoma protuberans and dermatofibroma: lack of correlation with proliferation rate. AB - p53 protein plays an important role in control of cell proliferation by suppressing proliferation of cells with DNA damage. Mutations of the p53 gene increase the stability of the encoded nonfunctional protein which accumulates in the nuclei, allowing it to be detectable by immunohistochemistry. Mutant p53 protein has been observed in preneoplastic and neoplastic conditions supporting its role in the development of some human cancers. In this immunohistochemical study, we examined p53 expression in 12 Dermatofibrosarcoma Protuberans (DFSP) and 10 Dermatofibromas (DF). Results were compared with the cellular proliferation rate by using the monoclonal antibody Mib-1 which detects Ki-67 antigen expression. Nuclear accumulation of the p53 protein was observed in 11 DFSP. All DF were negative for p53. No statistical correlation could be established between p53 and Mib-1 staining in our cases. We conclude that mutations of the p53 gene may be involved in the molecular pathogenesis of DFSP but not of DF. Mib-1 index can not be successfully used to distinguish DFSP from DF. PMID- 7499570 TI - A comparison of clinical, morphological and immunohistochemical features of lymphomatoid papulosis and primary cutaneous CD30(Ki-1)-positive anaplastic large cell lymphoma. AB - The advent of immunoperoxidase technique on paraffin embedded tissue has identified a number of shared immunologic markers present in various lymphoproliferative cutaneous disorders. Two such disorders are the recently described primary cutaneous CD30-positive anaplastic large cell lymphoma (ALCL) and lymphomatoid papulosis; both entities are characterized by CD30-positive large atypical cells predominantly of T cell origin. We have compared the clinical, morphological and immunohistochemical features of 50 patients with lymphomatoid papulosis to a group of 27 patients with cutaneous CD30-positive ALCL. There are clear differences between the clinical presentation in these two diseases, and although both are characterized by similar atypical cells, the histologic pattern and distribution of atypical cells is sufficiently different to allow distinction and specific diagnosis based on hematoxylin and eosin stained sections supported by the immunohistochemical stains. In addition, both diseases are characterized by a long benign course, rarely complicated by development of lymphoreticular malignancy and invariably demonstrate CD30 (Ki-1) antigen positive large atypical cells. PMID- 7499571 TI - Kimura's disease and angiolymphoid hyperplasia with eosinophilia: new observations from immunohistochemical studies of lymphocyte markers, endothelial antigens, and granulocyte proteins. AB - Kimura's disease (KD) typically presents as large subcutaneous masses in young Oriental males. It is characterized by deep inflammation with vascular proliferation, lymphocytic nodules with subcutaneous germinal centers, fibrosis, and edema. In comparison, angiolymphoid hyperplasia with eosinophilia (AHLE) occurs in all races and the lesions usually are smaller and more superficial. The causes of these two diseases are debated. We compared histologic features of 4 cases of KD with 22 cases of ALHE and studied expression of endothelial antigens and lymphocyte markers as well as localization of eosinophil, mast cell, and neutrophil granule proteins in lesional tissue. T-cell lymphoid aggregates with well-formed B-cell germinal centers occurred in KD, and nodular and diffuse T cell infiltration with small B-cell clusters occurred in ALHE. Endothelial proliferation was more pronounced in KD, lacking the atypical histiocytoid endothelial cells characteristic of ALHE. Many intact eosinophils infiltrated lesions in both diseases, although KD had less extracellular granule protein deposition than ALHE. Intact mast cells were seen in both diseases. There was neutrophil elastase staining of occasional scattered intact cells but no extracellular deposition. Compared with KD, ALHE is more varied in its clinical, histopathologic, and immunohistochemical features. PMID- 7499572 TI - Juvenile xanthogranuloma: a clinical, histopathologic and immunohistochemical study. AB - Juvenile xanthogranuloma (JXG) is a benign histiocytic proliferation of uncertain histogenesis which usually resolves spontaneously. Histopathologically, classic lesions are characterized by diffuse proliferations of foamy histiocytes, many of which may be multinucleated (Touton cells), admixed with lymphocytes and eosinophils. Histologic variants of JXG, perhaps representing evolving lesions, may lack these typical histopathological features, showing diffuse infiltrates of non-foamy mononuclear histiocytes without Touton cells, posing problems in differentiation from other histiocytic or melanocytic proliferations. Immunohistochemically, JXG is characterized by variable expressions of several histiocytic markers as well as the absence of staining for S100 protein. To assess better the spectrum of histopathological and immunohistochemical features of JXG, we studied nine cases of classic or histologic variant of JXG. The cases were evaluated by light microscopy and with an extensive battery of antibodies. All 9 cases, regardless of their light microscopic appearance, showed markedly positive staining with histiocytic markers including CD68, HAM56, cathepsin B and vimentin, but did not stain for S100 protein. Antibodies to factor XIIIa stained positively in 8 cases while staining for other markers was variable. Our results suggest that the histiocytes in JXG lesions have macrophagic differentiation, probably representing a reactive process to an unknown stimulus. PMID- 7499573 TI - Immunohistochemistry of dermatofibromas and benign fibrous histiocytomas. AB - Dermatofibromas (DF) are common, benign skin tumors composed predominantly of cells having elongated nuclei and very scant cytoplasm (i.e., fibroblasts) and capillaries in a collagenous stroma. Some authors distinguish DF from benign fibrous histiocytomas (BFH), which are composed of cells with round to oval nuclei and abundant cytoplasm (i.e., histiocytes). In general, this group of tumors expresses factor XIIIa but not the antigen recognized by MAC 387. However, immunohistochemical differences specifically between DF and BFH have not been reported. We have studied the immunophenotype of 23 lesions having morphologic features predominantly either of DF (17 cases) or BFH (6 cases) using antibodies against desmin (muscle marker), alpha-smooth-muscle actin (muscle and myofibroblast marker), CD68 and HAM56 antigen (markers commonly expressed by macrophages, so called "histiocytic" markers), CD34 (a marker present in hematopoietic, vascular, and occasional dermal dendritic cells), and factor XIIIa (a transglutaminase present in many cells including dermal dendrocytes). Many spindle-shaped cells expressed alpha-smooth-muscle actin while many large, round cells expressed the histiocytic markers. However, most lesions expressed at least focally both alpha-smooth-muscle actin and "histiocytic" markers. Thus a clear cut distinction between DF and BFH could not be made based on immunophenotype alone. Additionally, the prominent alpha-smooth-muscle actin immunoreactivity and desmin non-reactivity suggests myofibroblastic differentiation in the spindle cell regions of these tumors, and indicates that expression of alpha-smooth muscle actin cannot be used as definitive proof of muscle differentiation in spindle-cell tumors. We conclude that DF and BFH are not discrete entities, but represent polar expressions of one nosologic entity exhibiting both myofibroblastic and "histiocytic" differentiation. PMID- 7499574 TI - Halo nevus or halo phenomenon? A study of 142 cases. AB - One hundred and forty-two (142) halo nevi were reviewed. For 66 cases the diagnosis of halo nevus was made both clinically and pathologically, and for 76 cases the diagnosis was based on histological grounds alone. The nevi were classified by type and by degree of atypia. Of the 142 nevi, all were compound, junctional, or intradermal nevi except for one case of a Spitz nevus and two cases that could not be further classified. For those with a clinicopathological diagnosis of halo nevus, 11% exhibited moderate atypia; 16% exhibited minimal atypia to only focally moderate atypia; 24% minimal atypia; and 49% exhibited no significant atypia. For those cases where the diagnosis was pathological only, there was also a broad spectrum of atypia identified, with 8% exhibiting focally severe or severe atypia. This study supports the concept that the halo nevus should not be regarded as a single clinicopathological entity, but rather that the halo phenomenon occurs in a wide spectrum of nevus types exhibiting a wide spectrum of histological atypia. The pathologist is therefore encouraged to classify halo nevi on the basis of the nevus cell population alone, using whatever classification normally utilized. PMID- 7499575 TI - Basal cell carcinoma of the sole. AB - Basal cell carcinoma (BCC) of the plantar surface of the foot is rare, with only 22 previously reported cases. This clinicopathologic study is based on 20 cases of BCC of the plantar surface and plantar-like surfaces from adjacent lower lateral and medial aspects of the foot, submitted to a large podiatric laboratory from 1986 through June 1992 (total specimens for this period = 518,624; total BCC of lower extremities, below knee = 53). There were 15 women and 5 men. The average patient age was 73 years, with a range from 52 to 92 years. The duration of the lesion before diagnosis was 2 months to 12 years, with an average of 2 years. Three patients had a history of trauma. Podiatric clinical diagnoses included BCC (4), SCC (3), soft tissue tumor (2), nevus (1), granuloma (1), keratosis (2), verucca (1), and psoriasis (1). Follow-up information was available on 10 patients; all were free of disease up to 64 months, with an average follow-up of 15.7 months. Three of 20 BCC showed predominant histologic patterns characteristic of fibroepithelioma of Pinkus (FEP). An additional three BCC showed focal or suggestive patterns of FEP. Fourteen tumors showed ordinary BCC histologic patterns. No multicentric-superficial or morphea like BCC were observed. The relatively high incidence of FEP in BCC of the sole correlates with abundant sweat glands and lack of hair follicles on the plantar surface, in accordance with the recent proposal that FEP derives its histologic pattern from the spread of BCC down eccrine ducts, eventually replacing them with solid strands of tumor. PMID- 7499577 TI - Toxic epidermal necrolysis. A study of the sweat glands. AB - Toxic epidermal necrolysis (TEN) is a rare dermatological emergency which is caused by drugs and results in a high morbidity and a mortality rate of 20-30%. Ten confirmed cases of TEN were retrieved and the sweat glands were examined. Abnormalities of the sweat duct were found in all the cases and included basal vaculopathy, increased lymphocytic infiltration, basal apoptosis, basal cell hyperplasia and necrosis of the duct. Destruction and loss of the distal duct was present in 40% of the cases and was accompanied by dilatation of the proximal duct. In all cases, the proximal duct was involved to a lesser degree and in continuity with the distal duct, supporting the possibility of cytokine action. The secretory gland was normal. The presence of a significant reduction of sweat ducts in a disease process that involves 30-100% of the total body surface area is of clinical importance. PMID- 7499576 TI - Proliferative activity in metastatic malignant melanoma: comparison of pre treatment and post-treatment metastases. AB - In this study, the proliferative activity of malignant melanoma metastases was assessed before and after isolated limb perfusion chemotherapy by quantitating AgNORs, mitoses and PCNA activity. No significant difference in either AgNOR count, mitotic activity or PCNA index was observed. We conclude that AgNOR count, mitotic activity and PCNA index were not significantly effected by isolated limb perfusion chemotherapy. PMID- 7499578 TI - The use of Michel's transport medium for immunofluorescence and immunoelectron microscopy in autoimmune bullous diseases. AB - We report the use of Michel's solution, already a well established transport medium, in the combined use of direct immunofluorescence (IMF) and pre-embedding immunoelectron microscopy in 3 subepidermal bullous diseases--bullous pemphigoid (BP), epidermolysis bullosa acquisita (EBA) and dermal binding linear IgA disease (LABD). Our studies demonstrated that electron microscopy of normal skin maintained in Michel's medium for up to 28 days showed remarkable preservation of all components of the basement membrane zone, including the ultrastructure of the basal keratinocytes, dermoepidermal junction and papillary dermis. However, epidermal cell cytolysis occurred after just 48 hours. Immunoelectron microscopy using a gold probe has enabled us to localise the immunoreactants in bullous pemphigoid, epidermolysis bullosa acquisita and dermal-binding linear IgA bullous dermatoses. Our findings are comparable to and as equally reliable as those on immunoelectron microscopy of fresh skin biopsies with no loss of antigen deposition, and demonstrate an effective new use of a well established transport medium. PMID- 7499579 TI - Histologic findings in acute HIV exanthem. AB - We describe a patient with acute HIV exanthem with papulovesicular lesions and epidermal necrosis. We also review the literature regarding the histopathologic findings of acute HIV exanthem, which appears to be most commonly characterized by a perivascular lymphocytic infiltrate without epidermal change, but which may be associated with spongiosis, vacuolar alteration, or epidermal necrosis. PMID- 7499580 TI - Choriocarcinoma presenting as a cutaneous metastasis. AB - The rare presentation of choriocarcinoma as a cutaneous metastasis in a 23-year old male is reported. Histological and immunohistochemical analysis of the biopsy material demonstrated two distinct cell populations, syncytiotrophoblasts and cytotrophoblasts, with syncytiotrophoblasts strongly positive for human chorionic gonadotropin antigen. Subsequent clinical evaluation revealed a testicular tumor with metastases to lungs, brain, liver and kidney and increased serum levels of human chorionic gonadotropin. The patient died shortly after diagnosis due to complications of metastatic disease despite chemotherapy. PMID- 7499581 TI - Median raphe cyst of the penis with ciliated cells. AB - Cystic lesions occurring on the ventral surface of the penis have been classified as median raphe cysts of the penis. They are lined by pseudostratified, columnar or stratified squamous cell epithelium, mimicking the epithelial lining of the male urethra. Ciliated cysts of the human skin are unusual. Cystic lesions containing ciliated cells have been noted to occur in the chest, neck, or head, and bronchogenic origin has been the most accepted explanation for its origin. Other reports show the presence of ciliated cysts on the lower extremities, and the mechanism of formation is still a debated question. A case of median raphe cyst of the penis containing ciliated epithelium is presented. The existing literature about these cutaneous lesions is reviewed, including the possible mechanisms believed to be involved in its origin. PMID- 7499582 TI - Eruptive xanthomata with urate-like crystals. PMID- 7499583 TI - The three rules of life: some modest advice for those embarking on a career in dental research. PMID- 7499584 TI - Sumter Smith Arnim, DDS, PhD (1904-1990): a pioneer in preventive dentistry. AB - This article reviews the life story and contributions of Sumter Smith Arnim (1904 1990), a noted author, lecturer, teacher, and researcher in preventive dentistry. Following in the footsteps of Levi Spear Parmly, MD, DDS (1790-1859), and Charles C. Bass, MD (1875-1975), Arnim developed and popularized a wide range of preventive concepts and practices, especially dental plaque control via mechanical means. Today, these innovative ideas and techniques continue to have an impact on oral health research, the practice of dentistry, and the teaching of oral hygiene to dental patients. This article traces both the development of these oral health concepts and Arnim's study of dental plaque ("microcosm"), which he characterized by means of phase-contrast microscopy and cinemicrography. PMID- 7499585 TI - Effect of growth hormone on the distribution of decorin and biglycan during odontogenesis in the rat incisor. AB - Previous studies have shown that growth hormone can influence the expression of N acetylgalactosamine-containing molecules in the extracellular matrix of developing rat incisors. N-acetylgalactosamine is a principal component of proteoglycans containing chondroitin sulfate and dermatan sulfate, as well as of some glycoproteins. Since chondroitin sulfate proteoglycans are identifiable components in enamel, dentin, and cementum, we have tested the hypothesis that growth hormone modulates their expression in developing rat incisors. The distribution of the chondroitin-sulfate-rich proteoglycans, decorin and biglycan, was investigated. We used the Lewis dwarf rat as a model because their circulating growth hormone levels are markedly reduced. Polyclonal antibodies against decorin and biglycan were used to localize these two proteoglycans. Semi quantitative assessments of the staining patterns and intensities were made for each proteoglycan within compartments of the developing teeth. In normal Lewis rats, decorin and biglycan differentially expressed throughout the enamel organ, dental papilla, and dental follicle. Decorin displayed a wide distribution throughout all three regions and was closely associated with different cellular components. In contrast, biglycan showed little association with cells and was identified in the predentin and osteoid matrices. The expression of both proteoglycans was dramatically decreased in the growth-hormone-deficient animals. Administration of growth hormone to the dwarf rats markedly elevated the expression of both proteoglycans, approximating the distribution and intensity of staining seen in normal animals. These findings confirm that growth hormone status can modulate the expression of decorin and biglycan, and hence matrix deposition, in the rat tooth. PMID- 7499586 TI - The kinematic center: a reference for condylar movements. AB - The kinematic center of the temporomandibular condyle is that condylar point which follows as much as possible the same movement path during different types of mandibular movements. In this study, the location of the kinematic center with respect to the palpated lateral pole of the condyle was investigated. Also, the lengths of the condylar movement path reconstructed by means of the kinematic center and the palpated condyle were compared. Mandibular movements were recorded with 6 degrees of freedom in 20 healthy subjects. A software procedure calculated the location of the kinematic center as that mandibular point for which the protrusive and opening movement path showed a minimal difference. For each subject, its average location was calculated on the basis of 16 pairs of protrusive and opening movements. The kinematic center was located posteriorly and superiorly with respect to the palpated condylar point (p < 0.0001). The standard deviation in the anterior-posterior coordinate of the average kinematic center was smaller than that in the superior-inferior coordinate (p < 0.0001). During opening, the path length of the kinematic center is longer than that of the palpated lateral pole of the condyle (p < 0.0001). In contrast to left-right differences found in the path lengths of the lateral pole of the condyle, no left right differences were found for the kinematic center. PMID- 7499587 TI - Function-dependent anatomical parameters of rabbit masseter motor units. AB - Rabbit masseter motor units (22) were studied by stimulation of trigeminal motoneurons. We tested the hypotheses that masseter motor units facilitate fine motor control by concentrating fibers in small areas and that the distribution of motor unit fibers depends on the fiber type. The twitch contraction time and the isometric tetanic force were registered. The motor unit fibers were depleted of their glycogen by prolonged stimulation. Serial sections of the entire muscle were stained with the periodic acid Schiff (PAS) and monoclonal antibody stains. The muscle fibers of the motor unit were mapped and identified by four myosin heavy-chain (MHC) isoforms: I, IIA, IID, and cardiac-alpha. In the PAS-stained sections, anatomical parameters of the motor units, affecting the force output, were analyzed: the innervation ratio (IR), motor unit territory area (TA), and relative (R-DENS) and absolute (A-DENS) motor unit fiber densities. The fiber cross-sectional area (F-CSA) was measured for each MHC fiber type. The F-CSA sum of all motor unit fibers, the physiological cross-sectional area (P-CSA), was calculated. The IR ranged between 77 and 720 fibers (mean, 267). The mean TA was 8.71 mm2 (range, 4.45 to 19.58). The mean R-DENS was 10 fibers per 100; the A DENS was 31 fibers per mm2. Linear correlations were found between the IR and the R-DENS and between the tetanic force and the IR. The F-CSAs showed a stepwise increase in value from type I- to IID-MHC fibers. The mean P-CSA was 0.90 mm2 (range, 0.09 to 2.97). A high linear correlation was noted between the P-CSA and the tetanic force. In conclusion, increase of motor unit size expressed in higher fiber counts and forces is accomplished by increase of the fiber density. Thus, forces can be exerted selectively in restricted regions of the masseter muscle. Differences in fiber orientation due to complex muscle pinnation emphasize the possibility of an accurate muscle performance. PMID- 7499588 TI - The role of sensory information in the control of rhythmic open-close movements in humans. AB - Chewing requires a low level of muscle activity for jaw movement. Additional muscle activity is required to overcome the resistance of the food. The additional muscle activity consists of two contributions, an anticipating contribution before food contact and a peripherally induced contribution, about 23 ms after food contact. The amounts of both contributions depend on the information about food resistance obtained in preceding chewing cycles. It is not known whether this information is preserved if the resistance is absent during only a limited number of chewing cycles. Our aim was to investigate the extent to which information about food resistance obtained during chewing is used during subsequent cycles to generate anticipating and peripherally induced muscle activity. Subjects made rhythmic open-close movements at their natural chewing frequency, controlled by a metronome. Food resistance was simulated by an external force acting on the jaw in a downward direction during part of the closing movement. Jaw movement and surface EMG of the masseter and suprahyoid muscles were recorded during experiments in which sequences of at least 20 cycles with the force were alternated with a small, random number (from 1 to 10) of cycles without the force. The amount of anticipating muscle activity as well as the peripherally induced muscle activity in the first cycle with the force gradually decreased as a function of the number of preceding forceless cycles. About 30% of the additional muscle activity had an anticipatory origin, whereas the rest of the activity was evoked by the force regardless of the number of preceding forceless cycles. PMID- 7499589 TI - Absence of parasympathetic vasodilatation in cat dental pulp. AB - The existence and nature of parasympathetic nerve fibers in the dental pulp have long been a subject for discussion; indeed, vasodilator responses mediated by such nerve fibers have yet to be conclusively demonstrated in the dental pulp. This study was designed to determine whether parasympathetic vasodilator mechanisms do or do not exist in the cat dental pulp. Dynamic changes in pulpal blood flow (PBF), with mandibular lip blood flow (LBF) recorded as a control, were investigated in cat mandibular canine teeth by means of laser Doppler velocimetry. Peripheral trigeminal afferents (see below) were stimulated electrically to confirm that somato-parasympathetic reflex vasodilatation could be induced. The peripheral cut ends of the facial and glossopharyngeal nerve roots, which have been reported to contain parasympathetic nerve fibers to the oral tissues, were then stimulated intracranially. Electrical stimulation of trigeminal afferents (in the infraorbital nerve or the maxillary buccal gingiva) caused no change in PBF but did increase ipsilateral LBF. Neither facial nor glossopharyngeal nerve root stimulation caused a PBF increase, though both elicited increases in ipsilateral LBF. The vasodilator responses in the lip were sensitive to ganglion blockade (with hexamethonium), indicating vasodilatation via activation of parasympathetic vasodilator fibers. In contrast, intracranial stimulation of the trigeminal nerve root induced increases in both PBF and LBF which were reduced by pre-treatment with tripelennamine, indicating antidromic vasodilatation via the trigeminal sensory nerve. These results suggest that a parasympathetic vasodilator mechanism is not present in feline dental pulp. PMID- 7499591 TI - Tensile bond strength and SEM evaluation of caries-affected dentin using dentin adhesives. AB - Tensile bond strength measurements are commonly used for the evaluation of dentin adhesive systems. Most tests are performed using extracted non-carious human or bovine dentin. However, the adhesion of resins to caries-affected dentin is still unclear. The objectives of this study were to test the hypothesis that bonding to caries-affected dentin is inferior to bonding to normal dentin, and that the quality of the hybrid layer plays a major role in creating good adhesion. We used a micro-tensile bond strength test to compare test bond strengths made to either caries-affected dentin or normal dentin, using three commercial adhesive systems (All Bond 2, Scotchbond Multi-Purpose, and Clearfil Liner Bond II). For scanning electron microscopy, the polished interfaces between the adhesive bond and dentin were subjected to brief exposure to 10% phosphoric acid solution and 5% sodium hypochlorite, so that the quality of the hybrid layers could be observed. Bonding to normal dentin with either All Bond 2 (26.9 +/- 8.8 MPa) or Clearfil Liner Bond II (29.5 +/- 10.9 MPa) showed tensile bond strengths higher than those to caries affected dentin (13.0 +/- 3.6 MPa and 14.0 +/- 4.3 MPa, respectively). The tensile bond strengths obtained with Scotchbond Multi-Purpose were similar in normal and caries-affected dentin (20.3 +/- 5.5 MPa and 18.5 +/- 4.0 MPa, respectively). The hybrid layers created by All Bond 2 in normal dentin and by Clearfil Liner Bond II in normal or caries-affected dentin showed phosphoric acid and sodium hypochlorite resistance, whereas the hybrid layers created by All Bond 2 in caries-affected dentin and those created by Scotchbond Multi-Purpose to normal and caries-affected dentin showed partial susceptibility to the acid and sodium hypochlorite treatment. The results indicate that the strength of adhesion to dentin depends upon both the adhesive system used and the type of dentin. Moreover, the quality of the hybrid layer may not always contribute significantly to tensile bond strength. PMID- 7499590 TI - Urinary catecholamine levels in children with and without dental caries. AB - Urinary catecholamines have been used to measure emotionally stressful states which may affect the development of dental caries. This study investigates the hypothesis that children with and without dental caries do not differ significantly in the mean values of urinary catecholamines such as epinephrine, norepinephrine, and dopamine. As a test of this hypothesis, 314 children, males and females, aged from 6 to 8 years, were included in the study. Dental caries were recorded clinically and radiographically, and oral hygiene was evaluated by the recording of dental plaque. A 24-hour urine sample was collected for each subject, and a representative sample (25 mL) was analyzed by the HPLC technique to assay the catecholamine content. Socioeconomic factors such as parental age, education, and profession were recorded by a questionnaire distributed to the parents. Of the examined children, 38 (14 males and 24 females) were free of dental caries and constituted the case group. Two control groups, A and B, of 38 children each (14 males and 24 females) with dental caries were matched by age and gender. Differences in the quantitative and qualitative data were tested by the paired t test and the X2-test, respectively, while a regression analysis was applied to measure the effects of norepinephrine and dopamine on epinephrine. The logistic multiple-regression analysis was used to test, in the entire population, the impact of catecholamines and other related factors on the probability of subjects' developing dental caries. The 95% probability was used. The results showed statistically significant differences in epinephrine values between the case group and control groups A and B. The data suggest, therefore, that children with emotionally stressful states have higher probability of developing dental caries. PMID- 7499592 TI - Lipoteichoic acid inhibits remineralization of artificial subsurface lesions and surface-softened enamel. AB - Dental plaque produces not only acids by which underlying enamel is demineralized but also compounds which may inhibit repair of the lesions. The aim of this study was to determine how lipoteichoic acid, a bacterial compound that is abundant in dental plaque and inhibits calcium phosphate precipitation in vitro, affects the remineralization of incipient enamel lesions. Subsurface and surface-softened lesions were made in thin sections of bovine enamel, incubated with various amounts of lipoteichoic acid (isolated from Lactobacillus casei), and remineralized in 1.5 mmol/L CaCl2, 0.9 mmol/L KH2PO4, 130 mmol/L KCl, and 20 mmol/L Hepes, pH 7.0. Remineralization was followed during several weeks by repeated microradiography of the sections, and characterized by the changes in the integrated mineral loss of the lesions and the differential mineral profiles. The results showed that: (1) the effects of lipoteichoic acid on lesion remineralization were dose-dependent; (2) in subsurface lesions only the highest dose of lipoteichoic acid affected remineralization, which was delayed throughout the lesions; and (3) in surface-softened enamel, lipoteichoic acid did not affect the remineralization of the deeper parts, but remineralization of a surface zone of from 25 to 30 microns was increasingly inhibited with increasing doses. These effects were explained by different permeabilities of the surfaces of both types of lesions for the inhibitor: In subsurface lesions, lipoteichoic acid may have mainly clogged the porosities in the surface layer, whereas it could penetrate substantially into surface-softened enamel. PMID- 7499593 TI - Characterization of glucosyltransferaseB, GtfC, and GtfD in solution and on the surface of hydroxyapatite. AB - GlucosyltransferaseB, GtfC, and GtfD were purified by hydroxyapatite column chromatography, followed by ultrafiltration from the culture supernatant fluids of three Streptococcus milleri constructs (gift from Dr. H.K. Kuramitsu) which harbored individual gtf genes of Streptococcus mutans GS5. GtfB, GtfC, and GtfD were enzymatically active both in solution and in an experimental pellicle (HA CWS-Gtf) formed by adsorbing Gtf onto the surface of clarified human whole saliva (CWS)-coated hydroxyapatite (HA). The Km values for sucrose for all three enzymes were lower when the enzyme was adsorbed to a surface, compared with when it was in solution. In solution phase assays, and in the absence of primer dextran, glucan production was enhanced 75% when both GtfB and GtfD were present in the reaction mixture, compared with the sum of the individual enzyme activities (p < 0.005). This enhancement did not occur when GtfC was additionally present, or when the GtfB+GtfD enzyme pair was adsorbed onto HA-CWS. In additional experiments, glucan formed by GtfB or GtfC, but not by GtfD, on a HA-CWS-Gtf surface increased adherence of Streptococcus mutans GS5 and Streptococcus sobrinus 6715 by seven- to nine-fold compared with adherence when no glucan was present on the pellicle surface (p < 0.001). Further, treatment of the HA-CWS GtfB-glucan or HA-CWS-GtfC-glucan pellicle with alpha-1,6 dextranase significantly reduced adherence of both streptococcal strains (p < 0.001). These results show that GtfB, GtfC, and GtfD are enzymatically active in an adsorbed state and that the nature of their product glucan can influence the adherence of cariogenic oral streptococci to an experimental pellicle. PMID- 7499594 TI - Scanning electron microscope observations of CO2 laser effects on dental enamel. AB - Studies of the effects of carbon dioxide (CO2) lasers on dental enamel have demonstrated that surface changes can be produced at low fluences (< 10 J/cm2) if wavelengths are used which are efficiently absorbed by the hard tissues. In this study, scanning electron microscopy (SEM) was used to characterize the wavelength dependence of surface changes in dental enamel after exposure to an extensive range of CO2 laser conditions. Bovine and human enamel were irradiated by a tunable, pulsed CO2 laser (9.3, 9.6, 10.3, 10.6 microns), with 5, 25, or 100 pulses, at absorbed fluences of 2, 5, 10, or 20 J/cm2, and pulse widths of 50, 100, 200, 500 microseconds. SEM micrographs revealed evidence of melting, crystal fusion, and exfoliation in a wavelength-dependent manner. Crystal fusion occurred at absorbed fluences as low as 5 J/cm2 per pulse at 9.3, 9.6, and 10.3 microns, in contrast to no crystal fusion at 10.6 microns (< or = 20 J/cm2). Longer pulses at constant fluence conditions decreased the extent of surface melting and crystal fusion. The total number of laser pulses delivered to the tissue did not significantly affect surface changes as long as a minimum of 5 to 10 pulses was used. Within the four easily accessible wavelengths of the CO2 laser, there are dramatic differences in the observed surface changes of dental hard tissue. PMID- 7499595 TI - Duration of dentinal tubule occlusion formed by calcium phosphate precipitation method: in vitro evaluation using synthetic saliva. AB - The use of a calcium phosphate precipitation method occluded dentin tubules with apatitic mineral and, thus, showed good potential for the treatment of dentin hypersensitivity. The aim of this study was to elucidate the occluding behavior of the precipitate in the oral environment. Dentin disks treated by the calcium phosphate precipitation method, and disks treated with potassium oxalate, NaF, and SrCl2 solutions, were immersed in synthetic saliva, which was regularly replenished so that ionic concentration would be maintained. Treatment of dentin disks by the calcium phosphate precipitation method immediately reduced dentin permeability to 6 +/- 8%. When the disk was immersed in synthetic saliva, dentin permeability remained low, even seven days after immersion. Scanning electron microscopic observation showed no distinct boundary line between the precipitate and intertubular dentin; this indicated further mineralization on the precipitate. Potassium oxalate treatment also reduced the dentin permeability to 8 +/- 3%. However, the dentin permeability gradually but steadily increased with immersion time, reaching 39 +/- 14% at seven days. To elucidate the mechanism underlying dentin permeability changes in synthetic saliva, we immersed the precipitates, i.e., apatitic mineral and calcium oxalate, in a fixed volume of synthetic saliva. When calcium oxalate was immersed in synthetic saliva, there was a large concentration of oxalate ions, indicating dissolution of the calcium oxalate; this phenomenon was ascribed to the increase in dentin permeability. In contrast, calcium and phosphate ions decreased when apatitic powder, the precipitate formed by the calcium phosphate precipitation method, was immersed in synthetic salvia. The decrease in the calcium and phosphate ions in synthetic saliva indicated further precipitation of calcium phosphate on the apatitc precipitate.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7499596 TI - The site-specificity of supragingival calculus deposition on the lingual surfaces of the six permanent lower anterior teeth in humans and the effects of age, sex, gum-chewing habits, and the time since the last prophylaxis on calculus scores. AB - The hypotheses to be tested were: (i) that chewing sugar-free gum frequently and for long periods would be associated with higher amounts of supragingival calculus, and (ii) that there would be no site-specificity of calculus deposition on the lingual surfaces of the 6 lower anterior teeth. Subjects, 436 in Glasgow and 191 in Winnipeg, were scored for calculus at mesial, lingual, and distal sites on the lingual surface of each of the 6 lower anterior teeth, by the Volpe Manhold method. They also answered questions on the time since the last prophylaxis, the frequency of gum chewing, the type of gum chewed, and the length of a typical gum-chewing episode. A subset (233) of the Glasgow subjects were scaled and re-scored for calculus 3 months later. When the data for the logarithmic transformations of the initial calculus scores were subjected to stepwise multiple-regression analysis, the only factor which correlated significantly with initial calculus scores in both cities was the time since the last prophylaxis. In the Glasgow subjects scored 3 months after a prophylaxis, there was a negative correlation between chewing sugar-free gum and calculus scores, whereas in the Winnipeg subjects, age and the chewing of sucrose containing and sugar-free gum were positively correlated with calculus scores. Thus, the results were contradictory with respect to the first-tested hypothesis. The calculus distribution patterns were very similar in the subset of Glasgow subjects and the Winnipeg subjects, with the amounts on the lateral incisors and canines averaging 70.2% and 44.5%, respectively, of those on the central incisors. Thus, the second hypothesis was disproved. PMID- 7499597 TI - Clinical epidemiology is a needed basic science of clinical dentistry. PMID- 7499598 TI - Implications of hand dermatitis for dental education. PMID- 7499599 TI - Checklist agreement between standardized patients and faculty. AB - The agreement between standardized patients' (SPs) and faculty in scoring student performance is an important component in determining the accuracy of SP assessment scores. For this study, checklist scores completed by SPs were compared to checklists completed by faculty. The SPs completed the checklists immediately following each SP-student interaction. Faculty reviewed videotapes from randomly selected interactions and completed the same checklists as the SPs. Overall, agreement between SPs and faculty ranged between 81 percent and 92 percent, with a mean of 86 percent. The interpersonal skills agreement ranged from 78 percent to 97 percent and technical skills from 82 percent to 89 percent. Agreement between SPs and faculty was good especially in later cases. Discrepancies were most often due to SPs assigning credit when the faculty did not. This study supports the use of SPs as accurate, relatively inexpensive, and feasible recorders of student performance for selected skills. PMID- 7499600 TI - Dental hygienists' perceptions of preparation and importance of curriculum topics. PMID- 7499601 TI - Gender and the transition into practice. AB - Since 1990, over half the enrollees in advanced education pediatric dentistry programs have been women. The higher proportion of women in pediatric dentistry should permit examination of the practice patterns of groups of men and women at similar stages in their careers. In 1991 the American Academy of Pediatric Dentistry surveyed 4,950 dentists about a variety of issues related to practice patterns and demographics, obtaining 2,362 responses. This study conducted a secondary analysis of the survey data by developing three age-matched graduation cohorts based on gender and years since graduation: 1 to 5 years, 6 to 10 years, and over 10 years. Four areas were investigated: practice patterns, practice arrangements, distribution of time, and income. The overall differences in practice patterns between males and females were statistically significant for the Early Career Group (1 to 5 years). More males were in private practice and a higher proportion of them were practice owners. More women were dental school faculty or in private practice as an employee or contractor. The differences in practice patterns for males and females were not statistically significant for the Intermediate Career Group (6 to 10 years). In the Established Career group (over 10 years), the differences were again statistically significant, with more males as practice owners and shareholders and more women in solo practices. Analysis of time distribution showed that, in the two earlier career groups, women spend about twice as much time as men in child care. These findings may help to explain why many women in the early stages of their careers might prefer the flexibility of working for someone else. When the effects of gender and employment status on income were tested, significant differences were found for the Intermediate and Established groups, with males and practice owners having higher income levels.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7499602 TI - Child abuse and neglect curricula in North American dental schools. PMID- 7499603 TI - Knowledge, attitudes, and professional behaviors of third- and fourth-year dental students concerning AIDS and hepatitis B. PMID- 7499604 TI - Implementation of a tobacco cessation program in the student clinics. PMID- 7499605 TI - Nursing and the biological sciences. PMID- 7499606 TI - The cost of registered nurse-midwifery education in Malawi. AB - Educational institutions are challenged to produce more graduates with few resources in current sub-Saharan Africa. in such an environment, it is critical that cost information pertaining to nursing education be available to assist nursing education programmes in meeting the challenges of producing more nurse midwives with less cost. This study explores the cost of registered nurse midwifery education and describes factors which influence the cost of such education in Malawi. Results demonstrate that nursing education is costly, be it in a developing or develop country. The findings on cost of nurse-midwifery education have implications for policies related to the overall educational planning for the supply of registered nurse-midwives, the efficiency of nursing education programmes, the organization of nursing education curricula and the sharing of cost of education among the government, students, their families, and other sources. PMID- 7499607 TI - Post-natal depression: the relevance of sociological approaches. AB - Post-natal depression is much discussed yet definitions and approaches are not homogenous, neither in terms of the cause of post-partum mental ill health, its treatment or how further research in the area should proceed. This paper seeks to examine post-natal 'upsets' and to consider the different explanations that have been and could be made of post-partum mental ill health. The paper reviews the dominant biomedical and psychological approaches, evaluating their ability to explain post-natal mental illness. The writer believes biological and psychological approaches are in the ascendance but seeks to demonstrate that they do not present a full picture. Sociological approaches drawing upon stress, labelling and feminist models are examined, exploring new ways of looking at post natal illness. The paper concludes that biological and psychological approaches do not provide complete explanations and a multidisciplinary approach is needed. Most significantly the woman's own perception of post-natal ill health is largely absent from the literature. The need for an approach using ethnographic methods is highlighted. PMID- 7499608 TI - Drug-taking behaviours of pregnant substance abusers in treatment. AB - The purpose of this research was twofold: to describe the types of drugs and alcohol abused by pregnant multisubstance abusers enrolled in a substance abuse and treatment programme, and to describe the types of changes in drug-taking behaviours during pregnancy. An exploratory study, including interviews and chart reviews, was conducted with 31 pregnant multisubstance abusers. Drug-taking behaviours, demographic factors of age, ethnicity, martial status, education, and pregnancy characteristics of gravida, living children, level of prenatal care and trimester of pregnancy were measured. Most of the women were in their Twenties, Hispanic, single and had some high school education. Most of the women were multiparas to grand multiparas, yet they had only one or two living children, and all were receiving prenatal care. Their self-reports were validated by urine screening. Generally, the women reported that they did change their drug-taking behaviours, and many women decreased their substance abuse. PMID- 7499609 TI - Health-as-process: a health visiting perspective. AB - Health visitors are community nurses who define their role specifically in terms of health promotion, although they have often found difficulty in explaining how their seemingly diffuse and unfocused practice achieves this. The grounded theory study reported here set out to uncover some of the hidden processes and features embedded within health visiting, so the practice might be more clearly articulated. This paper reports one aspect of that study, which suggests that health visitors treat health as a lifelong process, involving the accumulation and use of 'resources for health'. The relevance of approaches which appeared both caring and educational will be set in the context of therapeutic nursing and adult education theories. Historically, health visitors have links with both nursing and health education; the analysis presented here helps to clarify the relationships between these different areas of work. The paper will explain how treating health as a process allows multiple competing views and ideas about health and health promotion to be integrated into a manageable form, thus allowing positive health to be promoted within a broad, acceptable socio-cultural context. However, the analysis also highlighted various points at which treating health as a process contradicts the firmly measurable requirements of the market orientated health service. Nevertheless, where health visitors are sufficiently skilled, and are permitted by their employing authorities to use this approach, it may serve to protect clients from intrusive and accusing interventions, made in the name of health promotion. PMID- 7499610 TI - Nursing, humanism and transcultural theory: the 'bracketing-out' of reality. AB - This paper addresses the emergence of a humanist discourse within nursing and questions the extent to which it represents the panacea implied within much of the humanist nursing literature. Particular attention will be given to whether it represents an ontological and epistemological framework capable of understanding the social, economic and political dynamics formative in the structuring of nurse client relations. It will be argued that the humanist analyses extant within much nursing literature are vague, idealistic, inconsistent and inadequate in the sense that they offer little by way of a meaningful analysis of power. A critique will also be made of the methodological individualism implicit within much humanist analyses. The paper will go on to identify the influence of humanist approaches on transcultural theory, and the manner in which the epistemological foundations of the latter have shared the limitations of the humanistic nursing approach generally. As such, the transcultural nursing literature is often vague, inconsistent in its use of terminology, lacking in any rigorous analysis of power, and suspect in its conceptualizations of culture. Its capacity for enabling nurses to examine critically the socio-economic and political dynamics of nurse-client relations and develop strategies for addressing racisms, considered by many to be endemic within nursing and health care system generally, is seriously undermined. PMID- 7499611 TI - Transcultural nursing: a perspective derived from Jean-Paul Sartre. AB - Transcultural nursing care is a well-established theoretical framework. This theory, developed by Madeleine Leininger, can have an impact on clinical practice, academic preparation and nursing research. The heightened interest in applying transcultural care and the increased need to espouse this framework have been triggered by constant population changes. This paper defines transcultural nursing care and expands its definition further by comparing and contrasting it with Jean-Paul Sartre's existentialist perspective. The two modes of existence as defined by Sartre are closely examined. Concrete examples are offered as cultural phenomena and the components of a cultural assessment are discussed and applied. The philosophical thesis is approached from a transcultural nursing perspective and existentialism is shown to serve as a foundation for transcultural nursing. The advantages and disadvantages of transcultural nursing practice and the preparation of transcultural nurses are finally addressed. PMID- 7499612 TI - Are nurses aware of the differing health care needs of Vietnamese patients? AB - This study examined the knowledge a small group of community nurses have of the Vietnamese culture. In our multi-cultural society nurses are being challenged to offer culturally appropriate care. Provision of care depends on an accurate assessment of health care needs which may differ from those of the dominant culture. The results indicated a lack of knowledge of the Vietnamese culture and the specific health care practices of the Vietnamese. This lack of knowledge is being rectified by in-service education. PMID- 7499613 TI - Challenging old notions of professionalism: how can nurses work with paraprofessional ethnic health workers? AB - Despite a recognition that health care systems in western countries need to become more responsive to their culturally diverse populations, health professionals have been slow to change their dominant monocultural work practices. Paraprofessional ethnic health workers have been employed in Australia to increase the access of non-English-speaking background communities to health services; however, they have generally not been able to form effective teamwork relationships with health professionals. Research into the use of ethnic health workers is explored to propose changes in the way that professional nurses practise alongside these workers. Concepts within professionalism and within a primary health care approach are considered against a framework of cultural differences between nurses and non-English-speaking background clients. It is unrealistic that nurses should expect to attain the cultural knowledge required to provide total care to all their clients in a diverse society, without a partnership with cultural intermediaries. Such an expectation is likely to produce lists of cultural traits that stereotype ethnic clients, rather than lead to an understanding of each client's individual needs. More effective teamwork between professional nurses and paraprofessional ethnic health workers is proposed so that together they can ensure health care is delivered in ways that are flexible and responsive to cultural differences. PMID- 7499614 TI - A review of the historical and social processes contributing to care and caregiving in Chinese families. AB - The purpose of this paper is to review the literature on caring and caregiving in Chinese families in relation to the contribution made by historical and social processes. The beliefs and traditions of care appearing in the professional and popular literature are explored to enable comparison with recent research from Hong Kong, China and the United Kingdom. Caring emerges as profoundly complicated and ambivalent, drawing on notions of morality, obligation, love, kinship and gender responsibility. Furthermore, caring in contemporary Chinese families in Hong Kong appears to have more similarities than differences with western families, possibly due to changing kinship networks. Such conclusions have particular relevance for nursing in the light of recent policy directives in Hong Kong promoting the role of the families in caring for their dependent members based on the assumption that families can and will care. PMID- 7499615 TI - Implementing change in nurses' professional behaviours: limitations of the cognitive approach. AB - This paper reports on a study of the effects of an intervention on a group of nurses' professional behaviours and on the process of behaviour change, according to the cognitive approach. The quasi-experimental design followed in the study resembles the time-series design. The study took place in a general state hospital in Lisboa and centred on the cardiology unit. Subjects were all nurses giving direct patient care (n = 17). The study extended over a period of 13 months. Results showed that the effects of the intervention were the adoption of some of the expected behaviours. Several questions were raised by the study, and the discussion points out some of the limitations of a cognitive approach to the understanding of the process of change of nurses' professional behaviours. PMID- 7499616 TI - Joint nursing appointments: a vehicle for influencing health care change. AB - With the rapid pace of health care change nurses need to organize and respond quickly to the debate on future health care policy. Nurses from practice and academia working together can capitalize on their unique knowledge and expertise to influence the development of consumer- and health-oriented policies. In the past joint appointments have served to unite nurses in the education and research domains. These appointments must evolve to encompass policy-focused activities that will enable nurses to make a significant contribution to the transformation of the health care delivery system. PMID- 7499617 TI - The clinical nurse specialist: a way forward? AB - The words clinical nurse specialist have been part of the nursing vocabulary for some time, but are these nurses specialized practitioners like many of us, or is there a larger role for them to play? This paper is a piece of reflective writing and is intended to further the debate concerning clinical nurse specialists. The paper examines clinical nurse specialists from an intensive care unit perspective. It looks at the intensive care unit and the wealth of under-utilized experience these environments contain. The paper examines the existing and envisaged roles of clinical nurse specialists, their education and utilization, and also the difficulties that they are likely to face in their evolution, both from within the nursing profession and from outside it. PMID- 7499618 TI - Definitions of quality of life: a taxonomy. AB - Quality of life is of central concern in evaluative research; improved quality of life is probably the most desirable outcome of all health care policies. However, definitions of quality of life are as numerous and inconsistent as the methods of assessing it. Stemming from a larger piece of work looking at the definition and measurement of quality of life, this paper highlights the lack of a consensus definition of quality of life by means of a taxonomy of definitions that emerge from the literature. The paper describes and gives examples of four main types of definition which make up the taxonomy: global (type I); component (type II); focused (type III); and combination definitions (type IV). In addition, an outline of factors influencing the definition of quality of life is given, and an alternative strategy for both defining and measuring the concept (the use of lay definitions) is suggested. PMID- 7499619 TI - Comparison of the overall quality of life in 50 long-term survivors of autologous and allogeneic bone marrow transplantation. AB - Fifty long-term survivors of bone marrow transplant (mean post-transplant time = 42.4 months) participated in a study examining their psychosocial adjustment and quality of life. Differences between patients who received an autologous marrow transplant and those who received an allogeneic marrow transplant were identified. Patients with an autologous transplant had mainly psychological difficulties in their post-transplant adaptation, whereas patients with allogeneic transplant developed more physical problems. Overall, their psychosocial adjustment was similar and comparable with other medical groups of patients. A quarter of both groups had failed to return to work/education and up to 9.5% had difficulty in carrying out daily tasks. Twenty per cent of the patients with autologous transplant had clinical signs of anxiety and 10% clinical signs of depression, whereas there was an incidence of 10% of patients with allogeneic transplant with anxiety, but no cases with clinical depression. Family relationships were found to be more integrated and lower in conflict compared with normal families. Quality of life has been described as good to excellent in most of the patients. Multiple regression analysis showed that physical symptomatology, vocational adjustment and depression are predictors of the degree of the patients' quality of life. PMID- 7499621 TI - Busy doing nothing: activity and interaction levels amongst differing populations of elderly patients. AB - This paper considers the activity and interaction levels amongst three differing populations of elderly patients (long-stay, short-stay and respite). It begins with a consideration of the literature on staff-patient interactions in care environments for elderly patients, highlighting the virtual absence of preplanned, purposeful activity. Data are then presented which suggest that, despite the emphasis nurses place on communicating with their patients, many patients continue to spend most of their time inactive. It is suggested that if the quality of care elderly patients receive is to improve, nursing staff must see the provision of activity as an integral part of their role and function. PMID- 7499620 TI - Survey of carer satisfaction with the quality of care delivered to in-patients suffering from dementia. AB - Quality assurance in British National Health Service provision stresses the importance of taking account of the consumer's viewpoint. Elderly patients with dementia are not always able to contribute usefully to satisfaction surveys. Therefore, their careers' views were sought in order to assess the quality of services offered to this client group. Forty-one careers of patients discharged from the eight wards for the elderly mentally ill in Leicestershire, England, were randomly selected. Individual focused interviews were conducted in careers' own homes. Both quantitative and qualitative data were obtained by use of a questionnaire designed to tap the patients'/careers' experiences from pre admission, through hospital stay to post-discharge. Interviews were asked to describe their favorable/unfavorable impressions of, and reactions to, all aspects of hospital care. These interviews were tape-recorded. Analysis of the data included quantitative measurements of scale ratings. Grounded theory was used to analyse qualitative data. A wealth of information was uncovered using this research technique. Much that was positive about the service was elicited. However, careers highlighted areas where they felt the quality of care could be improved within all the foci discussed. Twenty-two recommendations for quality improvements in service provision were made in the report as a result of this survey. PMID- 7499622 TI - The effect of living in a nursing home on socialization in elderly people. AB - The effects of institutionalization on elderly people are of significance both socially and physically. Currently, about 5% of elderly people in the United States live in long-term care institutions, although approximately 33% of all elders will be institutionalized for at least a short period of time. The effects on socialization of living in a nursing home have been studied and commented on from several perspectives. Several studies directed attention to the elderly person living in an institution. Others investigated the institution's response to residents' needs. Some studies have focused on attitudes of health care workers towards elderly people, and still others have examined the interaction between workers and residents. This review of the literature will first present studies focused on nursing home residents, then the institutions themselves, followed by studies of nursing home staff, and finally those focusing on the interaction between staff and residents. Discussion of possible directions for both practice and research will follow. The studies reported here were conducted in the United States, England, Northern Ireland and Finland. PMID- 7499623 TI - Attitudes and stereotypes: nurses' work with older people. AB - The study reported here sought to describe the differences between nurses' attitudes towards work with older people in the collective sense and their attitudes towards the individuals with whom they work. A convenience sample of 136 community nurses was obtained. Data were collected by means of a questionnaire incorporating the report of two critical incidents--one of effective practice and one of ineffective practice with older people. In addition in-depth qualitative interviews were conducted with 22 respondents. The findings lead the authors to challenge the common assumption that an identified lack of desire in nurses to work with older people is due solely to negative attitudes which nurses hold towards older people themselves. The paper concludes that the high dependency levels of older people and structure of nursing work with older people in hospitals means that fewer nurses make this area a positive career choice. PMID- 7499624 TI - 'To call it work somehow demeans it': the social construction of talk in the care of terminally ill patients. AB - The nurse has an important role in helping the terminally ill patient come to terms with the imminence of death. Such work is highly demanding and often stressful, but is work which is accorded a high moral priority by respondents in the study reported in this paper. The paper explores the ways in which nurses work to respond to patients' expressed psychosocial problems and the emphasis that they placed on providing opportunities for patients to speak about their impending death. PMID- 7499625 TI - Sleep disturbance in mid-life women. AB - A correlational field survey was used to describe sleep disturbance and explore the relationship of that sleep disturbance to the psychological and menopausal status of 23 mid-life women. Subjects were 40-55 years of age and had self reported sleep problems. Instruments included the sleep disturbance questionnaire (SDQ), Centre for Epidemiologic Studies depression scale (CES-D), Speilberger state-trait anxiety scale (STAI), a demographic form, 4 cc blood sample for follicle stimulating hormone analysis, and a 7-night sleep diary ('self report of sleep', or SRS). Ten of the 23 subjects reported periodic limb movement on the SDQ. Sleep diaries (SRS) revealed a characteristic sleep disturbance among 13 of the 23 subjects, which paralleled in some respects the changes in sleep efficiency observed with advancing age, but was more pronounced and was present at an earlier age. There was no statistically significant relationship between sleep disturbance and menopausal status or symptoms, or anxiety and depression among subjects, as shown by correlational coefficient and chi-square statistical tests. Results suggest that a characteristic sleep disorder accompanied by periodic limb movement may be responsible for sleep disturbance in mid-life women. What remains unclear is whether this sleep disturbance is related to aspects of the perimenopause not addressed in this study or to other aspects of ageing. PMID- 7499627 TI - The moderating effect of demographic variables on coping effectiveness. AB - In spite of the theoretical recognition regarding the role of coping as a moderator, empirical findings indicate that it has only a modest effect as such. This study was designed, therefore, to examine the moderating effect of demographic variables (DV) on coping effectiveness, i.e. active cognitive coping, avoidance coping and active behavioural coping. The study sample was comprised of 12 male and 65 female patients with rheumatoid arthritis who were treated in three outpatient clinics of large hospitals in Israel. Findings suggest that demographic variables do play an important, and somewhat surprising, role in the effectiveness of coping strategies to temper psychological distress. Of the various demographic variables studied, marital status was found to have a significant effect on active cognitive coping and avoidance coping, and adherence to a religious belief system was found to have a significant effect on avoidance coping. The implications of the results for coping effectiveness are discussed. PMID- 7499628 TI - Nurses' perceptions of the impact of continuing professional education on the quality of nursing care. AB - The purpose of this study was to understand what impact nurses perceived continuing professional education (CPE) to have on the quality of nursing care. Given that CPE will become mandatory once the necessary legislation has been implemented, the value and worth of CPE with regard to the quality of nursing care, in both a consumer-led and financially-driven NHS, merits serious consideration. Little research has been conducted into the real impact that CPE has on the quality of nursing care and this paper begins by providing an overview of the current debate within the profession. The method chosen for the study was qualitative and 18 nurses from a large hospital in the South of England were interviewed. The data were analysed using the grounded theory method and three categories emerged. The categories are described in detail and reveal some of the problems with which the nursing profession is faced. The study demonstrates how issues of finance were particularly high, with service managers sometimes negating nurses' CPE needs. However, the study reveals the real importance that nurses attached to CPE in supporting their professional status and the real impact that CPE and knowledge have on professional competence and the quality of patient care. PMID- 7499626 TI - Parents' perceptions of caring for an infant or toddler with diabetes. AB - Although the incidence rate of diabetes in children under the age of 3 years is increasing dramatically, no previous studies provide information about the day-to day experience of caring for these infants and toddlers. Because a young child with diabetes is dependent on the parent for his/her very existence, the purpose of this phenomenological study was to gain knowledge and understanding of the parents' experiences so that appropriate interventions could be developed and implemented to support parental care for this unique population. Findings revealed three distinct phases in the parents' experiences: the diagnosis and child's hospitalization; adjusting to care at home; and long-term adaptation. Within these phases, parents described inordinate amounts of stress exacerbated by the child's young age and the complex, intrusive nature of diabetes management. On the basis of these findings, implications for practice are discussed. PMID- 7499629 TI - A sharing in critical thought by nursing faculty. AB - A critical analysis of nurse education programmes has revealed an overt and covert curriculum; the overt being the one underpinned by values which espouse humanism and critical thought; the covert being the one which reflects the patriarchal system and is directed by a means-end rationality. In response to this dilemma the 'curriculum revolution' mandate for change, which occurred in the latter half of the last decade, called for nurse educators to unveil, understand and criticize the assumptions and values which guided their practice, so that they, and consequently their students, could be more responsive to the needs of society, value subjective experience, acknowledge theoretical pluralism, and share an egalitarian relationship. This paper explores the formation of a 'critical collective' of nurse academics who came together believing that, 'if you always do what you've always done, you'll always get what you've already got'. The major concern of this collective was to facilitate change within their work environment, through the development of strategies, so that the ideas of the 'revolution' were not lost to the mere rhetoric of curriculum documents. PMID- 7499630 TI - How male nursing students learn to care. AB - the current definitions of caring in nursing education denote a mutual relationship between teachers and students in which they interact to provide an environment which supports the students' personal and professional development. The learning of caring in an interactional context is typical of the female experience. It is unknown if this is representative of the male experience. The lived experience of 20 male students as they learned to care as nurses was the focus of this phenomenological research study. The participants concurred that, although caring as a nurse could be learned if one was 'open to learning', teachers could only facilitate this learning; they could not directly teach it. The interactional strategies of storytelling, modelling, being cared for, the 'aha' encounter, and observing and giving care, were used by the participants in learning to care as nurses. The implications of these findings for nursing education and research are presented. PMID- 7499631 TI - Nursing research in France: INSERM conference held in Paris, France, 10-11 March 1995. PMID- 7499632 TI - Inconsistent contraceptive use among female college students: implications for intervention. AB - The authors evaluated the longitudinal consistency of use of contraceptive and barrier methods among female college students attending a student health clinic. The majority of students using unreliable methods of contraception during follow up did not report unreliable contraceptive use at the baseline observation. Of 213 women who participated, 33% used unreliable contraception; 9% only at baseline, 14% only during 6-month follow-up, and 9% at both. Selection of a contraceptive method varied considerably over time. Of those using the oral contraceptive pill at baseline, 31% did not use this method during follow-up, and 43% of those using condoms at baseline had intercourse without a condom during follow-up. Single observations are inadequate to detect unreliable contraceptive use in this population; cross-sectional studies will fail to detect important transitions in contraceptive use. Interventions to prevent pregnancy and sexually transmitted disease among female college students should include a focus on the consistency of contraceptive use. PMID- 7499633 TI - Encouraging discussion of psychosocial issues at student health visits. AB - In a pilot study, the authors distributed a brief questionnaire dealing with psychosocial concerns to 200 students awaiting treatment in the student health service at a large urban university. They hypothesized that a questionnaire administered immediately before students visited the physician or family nurse practitioner would encourage the students to discuss problems such as anxiety and depression during the medical session. Using the questionnaire, they found, led to the practitioner's discovery of a condition requiring treatment or explaining a problem the student had not discussed in previous visits. Such a questionnaire, the authors suggest, could be a valuable means for identifying psychosocial concerns significant enough to require treatment or further evaluation. PMID- 7499634 TI - Propensity for seeking counseling services: a comparison of Asian and American undergraduates. AB - The authors used the somaticization scale of the Brief Symptom Inventory to study Asian and American college students' propensity for seeking counseling at an American university. They found that the Asian students were more likely than the American students were to report that they would seek counseling services when they experience somatic discomfort. The Asian students scored significantly higher on inclination for seeking counseling than did the American students. PMID- 7499635 TI - Values, participatory democracy, and healthcare resource allocation: an application to a campus community. AB - As healthcare reform is debated, it is important that decision makers consider the values of all citizens, that is, what people find desirable or useful about healthcare services. Several states have used town meetings in an effort to determine their citizens' views on the values in the realm of healthcare. In this article, the authors describe a process in which individuals actively participate in an open discussion about issues surrounding allocation of healthcare resources in a university setting. Three different groups from the university community participated in separate, structured meetings to discuss their values concerning the allocation of scarce healthcare resources. Such meetings give participants opportunities to learn about their values and those of other persons in the community. PMID- 7499636 TI - Student opinion leaders and HIV/AIDS knowledge and risk behavior. AB - This research applied the construct of opinion leadership to the health area of AIDS. Approximately 290 college students completed a questionnaire that identified individuals whom peers tended to use as sources of information about AIDS. The results showed that, among men, the AIDS opinion leaders tended to have a larger number of sexual partners than their nonleader counterparts and that they were no more likely to practice safer sex or be more knowledgeable about safer sex. Female AIDS opinion leaders tended to engage in less sexual activity than their nonleader counterparts, but showed a greater reluctance to communicate with men about condom use. In addition, female AIDS opinion leaders were neither more knowledgeable about safer sex nor more likely to engage in safer sex than were their nonleader counterparts. PMID- 7499637 TI - Presentation of unrecognized attention deficit hyperactivity disorder in college students. AB - Mental health providers at university counseling and health centers should be alert to the possibility that college students with previously unrecognized attention deficit hyperactivity disorder (ADHD) may present for initial evaluation at their centers. This study was a systematic chart review of 42 students at an Upper Midwest university who were diagnosed with ADHD during calendar year 1993. Diagnoses were made by the treating psychiatrist, who reviewed records for presenting problems; recent associated problems; previous evaluations as a child, adolescent, or adult; and associated problems in childhood. Presenting problems included ADHD symptoms, mood symptoms, nonspecific learning disability, and academic underachievement. Associated problems were depressive disorders, anxiety disorders, drug and alcohol abuse or both, dependency, legal problems, learning disabilities, and eating disorders. Thirty three percent had been evaluated for academic or behavior problems as children, and 36% had sought previous psychological care for non-ADHD symptoms as adults. Thirty-one percent were presenting at the university health center for their first evaluation. Childhood histories showed educational underachievement, learning disabilities, and behavior problems. PMID- 7499638 TI - Protocols constructed around the nursing process. 3: Autonomic dysreflexia. AB - Autonomic dysreflexia is a serious problem in patients with spinal cord lesions above the fifth thoracic level. This nursing protocol outlines assessment, diagnosis, and interventions for nurses and physicians and lists appropriate medications. PMID- 7499639 TI - Comments on the asthma protocol. PMID- 7499640 TI - The disruptive college student: some thoughts and considerations. PMID- 7499641 TI - Follow the leader. PMID- 7499642 TI - AGD/ADA. PMID- 7499643 TI - Post-treatment bacteremia. PMID- 7499645 TI - ADA and AAOM produce HIV treatment resource. PMID- 7499644 TI - Prefabricated overdentures. PMID- 7499646 TI - Dental amalgam and cognitive function in older women: findings from the Nun Study. AB - The authors determined the number and surface area of occlusal dental amalgams in a group of 129 Roman Catholic sisters who were 75 to 102 years of age. Findings from this study of women with relatively homogeneous adult lifestyles and environments suggest that existing amalgams are not associated with lower performance on eight different tests of cognitive function. PMID- 7499647 TI - Factors contributing to mercury exposure in dentists. AB - Although mercury exposure levels among dentists have been steadily decreasing, occupational mercury exposure among dental professionals continues to be a matter of interest. This study examined personal professional and office characteristics of dentists to determine which factors contribute most to exposure. The resulting information will help dental professionals change their practices to minimize unnecessary exposure to elemental mercury. PMID- 7499648 TI - Are gingival and periodontal conditions related to salivary gland flow rates in healthy individuals? AB - Some have suggested that gingival and periodontal health is related to salivary gland function; however, there are few data to support this hypothesis. The purpose of this study was to determine if correlations existed between major salivary gland flow rates and gingival and periodontal conditions in people of different ages. The results suggest that there is no consistent relationship between major salivary gland flow rates and gingival and periodontal conditions in healthy people. PMID- 7499649 TI - Clinical evaluation of restored maxillary incisors: veneers vs. PFM crowns. AB - This cross-sectional study compared the periodontal health and clinical acceptability of maxillary incisors restored with porcelain veneers vs. porcelain fused-to-metal crowns. The authors evaluated 60 patients, a total of 120 restorations, in groups according to the age of the restorations (zero to 60 months). Margins of veneers were generally more supragingival with less gingival inflammation. All veneers were clinically acceptable, while 5 percent of PFMs failed because of secondary caries. PMID- 7499650 TI - Assessing the duration of mandibular soft tissue anesthesia. AB - Dentists often employ solutions of 3 percent mepivacaine or 4 percent prilocaine without a vasoconstrictor in pediatric patients in an attempt to reduce the duration of mandibular soft tissue anesthesia. The authors compared the time course of soft tissue anesthesia produced by these solutions with that of 2 percent lidocaine plus 1:100,000 epinephrine in 60 adults. They found no reduction in the duration of soft tissue anesthesia when employing 3 percent mepivacaine or 4 percent prilocaine instead of 2 percent lido-epi. Combining these observations with local anesthetic dosage considerations, the authors recommend that 2 percent lido-epi be used when performing mandibular block injections in young children. PMID- 7499653 TI - What is restraint of trade? A review of the nation's antitrust laws. PMID- 7499651 TI - Sialorrhea and gastroesophageal reflux. AB - It is not unusual for patients who are pregnant or have a hiatal hernia to develop gatroesophageal reflux with the resultant symptoms of heartburn. This article reviews the role reflux plays in causing episodes of increased salivation. Stimulation of the esophagus by gastric acids excites and esophagosalivary reflex. A marked increase in salivary flow ensues, neutralizing the acid content of the esophagus. The dental practitioner is in a strategic position to be consulted about or recognize the existence of the esophagosalivary reflex and the consequent bouts of sialorrhea. Recognition mandates medical consultation. PMID- 7499652 TI - A new technique for restoration of worn anterior teeth--1995. PMID- 7499654 TI - A survey of ethical dilemmas and practical problems encountered by practicing dentists. AB - Dental practice has been subject to many changes in recent years, including third party involvement, infection control requirements and a rise in litigation. The authors conducted a broad-based national survey of practitioners to learn their views on the ethical and practical problems facing today's dentist. PMID- 7499655 TI - Public attitudes toward dentists: a U.S. household survey. AB - This article reports on research that assessed public attitudes toward dentists and the dental profession and compared them to dentists' own attitudes about their roles. Enhanced knowledge of patients' perceptions can help practicing dentists gain a better understanding of public perceptions of themselves and of dentistry as a profession, as well as enhance their relationships with patients. PMID- 7499656 TI - Who will speak for dentistry? PMID- 7499657 TI - What is a conspiracy? PMID- 7499659 TI - Direct reimbursement. PMID- 7499658 TI - Direct reimbursement, dental service plans. PMID- 7499660 TI - Sodium fluoride makes headway in osteoporosis study. PMID- 7499661 TI - Cigarette packaging appeals to teens. PMID- 7499662 TI - Risk of fluorosis in a fluoridated population. Implications for the dentist and hygienist. AB - The prevalence of enamel fluorosis has increased in optimally fluoridated areas in recent years. This has led to efforts to identify the cause or causes and to make recommendations that seek to maintain the caries-preventive effectiveness of fluoride use while minimizing the risk of fluorosis. In this study, the author estimated the potential direct impact that dental practitioners could have on reducing the amount of enamel fluorosis in U.S. children. The findings suggest that dental practitioners could have an important impact on reducing the prevalence of enamel fluorosis by guiding the public toward the most appropriate use of fluoride products. PMID- 7499663 TI - Infants' fluoride ingestion from water, supplements and dentifrice. AB - Concerns about dental fluorosis and the paucity of detailed fluoride intake data prompted this longitudinal study of fluoride intake in infants from birth to 9 months of age. On average, water fluoride intake greatly exceeded that from dietary fluoride supplements or fluoride dentifrice. However, fluoride supplements and dentifrice contributed substantial proportions of fluoride intake among children using them. Some children had estimated fluoride intake from water, supplements and dentifrice that exceeded the recommended "optimal" intake (a level that has yet to be determined scientifically). Practitioners should estimate fluoride ingestion from all these sources if considering systemic fluoride supplementation. PMID- 7499665 TI - Managing patients who have seizure disorders: dental and medical issues. AB - Some dental professionals may have concerns about treating patients who have seizure disorders. The authors contend that increased knowledge about seizures and how to manage them may make dental professionals more comfortable. This article reviews the various seizure types, discusses commonly used anticonvulsants and their side effects and outlines some special concerns dentists may have in providing care to these patients. PMID- 7499664 TI - Reducing bacterial aerosol contamination with a chlorhexidine gluconate pre rinse. AB - The authors compared the effects of chlorhexidine gluconate, an antiseptic mouthwash with essential oils and water on the bacterial aerosol contamination generated by an air polishing device. Patients rinsed with one of the three solutions before treatment. Bacterial counts collected during the treatment indicate that the chlorhexidine pretreatment rinse was significantly more effective than the other solutions in reducing bacterial aerosols. PMID- 7499666 TI - Preliminary estimates of the incidence and consequences of tooth fracture. AB - The authors studied all tooth fractures identified within a two-week period among adult enrollees in a dental health maintenance organization to determine the incidence and severity of this condition. They found complete fracture rates of 5.0 and 4.4 per 100 adults per year for all teeth and for posterior teeth, respectively, with 15 percent of fractures resulting in pulpal involvement or extraction. These estimates, the first to be reported, provide information that is potentially useful to patients and dentists making decisions about treatment intended to prevent tooth fracture. PMID- 7499667 TI - A case report. Ectopic molars that cause osteomeatal complex obstruction. AB - During a routine dental examination, a general dentist noticed facial asymmetry in a 42-year-old man. Dental radiography suggested a pathological condition in the sinuses. The dentist referred the patient to a head and neck surgeon for further examination. Computed tomography demonstrated ectopic molars in the maxillary sinuses blocking drainage of the sinuses. PMID- 7499668 TI - Assessment of periodontal status with PSR and traditional clinical periodontal examination. AB - The system of peridontal screening and recording was developed by the American Dental Association and the American Academy of Periodontology to simplify early detection of periodontal pathology. The authors evaluated the results of PSR against those of a conventional periodontal examination in 24 subjects, each of whom had a minimum of 18 natural teeth. The results of this study suggest that the PSR examination, while a useful screening tool, could be improved to further enhance its identification of patients with peridontal diseases. PMID- 7499669 TI - Abnormal occlusal conditions: a forgotten part of dentistry. PMID- 7499671 TI - Insurance exemption to the antitrust laws. PMID- 7499670 TI - EDI: novocaine for managing your dental claims. AB - According to the author, submission of dental claims through electronic data interchange technology is a cost-effective way for dentists to handle some of the administrative aspects of managing their practice. Understanding what's involved can take you a long way in your decision to submit dental claims electronically now and in the future. The author outlines the advantages of EDI and gives tips on how to incorporate EDI into your office. PMID- 7499673 TI - Studies on the relationship between the level of specific IgE antibodies and the clinical expression of allergy: I. Definition of levels distinguishing patients with symptomatic from patients with asymptomatic allergy to common aeroallergens. AB - BACKGROUND: The detection of specific IgE antibodies to environmental allergens does not always coincide with a diagnosis of clinically evident allergic disease, because some patients with positive skin and/or in vitro test results have no symptoms related to the allergen or allergens that induced the antibodies. OBJECTIVE: In a multicenter study the optimal cutoff values for specific IgE antibody levels and skin test results that could discriminate between patients with symptomatic and those with asymptomatic allergy were determined. METHODS: IgE antibodies specific for a panel of common aeroallergens were assayed with the Pharmacia CAP System (Pharmacia, Uppsala, Sweden) in two groups of patients, a group of 267 patients with symptomatic allergy and a group of 232 with asymptomatic allergy--both with positive skin prick test results--and in a group of 243 healthy, nonallergic control subjects. The cutoff values were established by receiver operating characteristic analysis. RESULTS: A significantly higher mean specific IgE antibody value was found in patients with symptomatic allergy compared with patients with asymptomatic allergy (p < 0.001) and in patients with symptomatic allergy compared with healthy control subjects (p < 0.001). The optimal CAP System cutoff value between patients with symptomatic and those with asymptomatic allergy was 11.7 kU/L, and when seasonal allergens were compared with perennial allergens, the cutoffs were 10.7 kU/L and 8.4 kU/L, respectively. The optimal cutoff value for the skin prick test was a wheel area of 32 mm2 for seasonal allergens and 31 mm2 for perennial allergens. The skin test had a lower diagnostic value (sum of sensitivity and specificity) than the CAP System. CONCLUSIONS: Cutoff values for specific serum IgE antibody levels are likely to be useful in clinical practice to distinguish symptomatic from asymptomatic allergy in patients with positive skin test results. PMID- 7499672 TI - Dose-related decrease in bone density among asthmatic patients treated with inhaled corticosteroids. AB - BACKGROUND: Inhaled corticosteroids are being prescribed more commonly and in higher doses than previously in the management of asthma. Although these topically active compounds have less potential for systemic impact than oral steroids, biochemical markers suggest that they are not devoid of systemic side effects. We conducted this study to investigate the effect of commonly prescribed doses of inhaled steroids on bone density. METHODS: We studied 36 patients with asthma. Those in group A (n = 18) had been taking inhaled beclomethasone dipropionate or budesonide in a dosage of 800 micrograms or more per day for at least 1 year. Those in group B (n = 18) had used only bronchodilator therapy. Adrenal function was assessed by morning serum cortisol level and by short adrenocorticotropic hormone stimulation test. Bone turnover was assessed by measurement of serum osteocalcin, alkaline phosphatase, and urinary pyridinium cross-links. Bone mineral density was measured by dual-energy x-ray absorptiometry with a Hologic QDR-1000 densitometer (Hologic Inc., Waltham, Mass.). RESULTS: Group A, mean age (SD) = 36.6 (8.4) years, had used inhaled corticosteroids at a mean dose of 1323 micrograms/day (range, 800 to 2000 micrograms/day) for a median duration of 24 months. Group B, mean age (SD) = 33.4 (8.1) years, had not been taking any form of steroid. Four patients from group A had suppressed morning serum cortisol; three of these had abnormal adrenocorticotropic hormone stimulation test results. All patients in group B had normal baseline adrenal function and an appropriate response to adrenocorticotropic hormone. Mean serum osteocalcin level in group A was significantly lower than that in group B (8.8 vs 14.2 ng/ml, p = 0.0003). Bone density measurements showed parallel changes: in group A the mean Z score (SD) of the femoral neck was -0.78 (1.02), significantly below predicted normal values (p = 0.0025). Mean Z scores of the lumbar spine and of femoral Ward's triangle were not significantly reduced. In group B the mean Z scores of the lumbar spine, femoral neck, and femoral Ward's triangle were all within normal limits. In group A the dose duration of inhaled corticosteroid therapy corrected for body mass index correlated negatively with bone density and adrenal function measurements. CONCLUSION: We conclude that the regular use of conventional doses of inhaled corticosteroids by patients with asthma can suppress adrenal function and decrease bone density in a dose-related fashion. PMID- 7499674 TI - The accuracy of features in the clinical history for predicting atopic sensitization to airborne allergens in children. AB - BACKGROUND: The clinical history is given considerable weight when one decides which allergens are responsible for a patient's symptoms, and in research studies the clinical history has been used as the "gold standard" with which different tests for allergy are compared. OBJECTIVES: To determine whether standardized questions accurately predict replies to detailed questions asked by an experienced allergist, and to assess the utility of certain standardized questions for predicting sensitization to individual allergen groups. METHODS: Trained interviewers put standardized questions to parents of 1160 children, aged 1 to 17 years, who had respiratory symptoms and had been newly referred to the allergy clinic of a children's hospital. For the first 151 of the subjects the answers were compared with those elicited by questions asked by a pediatric allergist. Skin prick tests and pollen counts were performed by a technologist. RESULTS: The standardized questions had an accuracy for predicting the allergist's history of 93% to 97% for all questions except one. The standardized questions with the highest accuracy for predicting the skin test results to the appropriate allergens were the following: for mite, improvement in symptoms when outdoors (66.8%) and when in dry areas 69.4%), and aggravation during house cleaning (65.9%) and when bed making (70.6%); for dog, symptoms when with dogs (80.6%); for cat, symptoms when with cats (77.3%); for tree pollen, symptoms worse in April (70.8%) and when among trees in March and April (80.8%); and for grass pollen, exacerbation in June (69.2l%) and during lawn mowing (71.2%). Although specificity was generally above 80%, sensitivity was variable, ranging from 11% to 56%. CONCLUSIONS: The standardized questions accurately predicted a detailed history obtained by an experienced allergist. Because standardized questions are reproducible they are the preferred method of history taking for research projects. Because several of the standardized questions have a high specificity they are useful for excluding sensitization to individual allergen groups, but because they have only a modest sensitivity, they are less helpful for detecting those who are sensitized to individual allergen groups. PMID- 7499675 TI - Ephedrine-saline nasal wash in allergic rhinitis. AB - BACKGROUND: Ephedrine nasal wash (0.25% to 1%) was once used in the treatment of sinusitis and other nasal and paranasal symptoms, but there are no reported controlled trials in allergic rhinitis. METHODS: In a blinded study, 118 patients with perennial allergic rhinitis were randomly divided into two groups of 65 and 53 patients, respectively, and administered either a placebo or a 1% ephedrine saline nasal wash once every 48 hours for a period of 4 weeks. These groups were then crossed over after a 4-week interval, and their weekly symptom scores and peak nasal inspiratory flow rates were monitored. RESULTS: Use of ephedrine saline nasal wash resulted in significant improvement in symptom scores and peak nasal inspiratory flow rates as compared with placebo (p < or = 0.001). A statistically significant difference between the two groups began in the second week. The effects of treatment lasted for more than 2 weeks after the study, without notable side effects, in most patients. CONCLUSION: A 1% ephedrine-saline nasal wash is an inexpensive and effective treatment for allergic rhinitis. PMID- 7499676 TI - Do subjects investigated for occupational asthma through serial peak expiratory flow measurements falsify their results? AB - BACKGROUND AND AIM: Serial assessment of peak expiratory flow (PEF) rates has been advocated as a sensitive and specific means of investigating occupational asthma. The possibility that, for several reasons, subjects do not accurately report their values has been raised. The availability of portable instruments that assess PEF and store timings and values now make it possible to estimate compliance and accuracy of results. METHODS: Twenty-one subjects consecutively investigated for occupational asthma were asked to assess their PEF every 2 hours during the day, both at work and away from work, with a VMX instrument (Clement Clarke International, Columbus, Ohio) and record the times and values on a sheet of paper. The subjects were not aware that the data were also being stored on a computer chip. The diagnosis was occupational asthma in eight subjects, personal asthma in four subjects, and neither condition in nine subjects. RESULTS: The mean duration of recording was 36 days (range, 14 to 79 days). At least 6048 values should have been recorded, but only 4839 (80%) were either recorded or stored. Reported values corresponded precisely to stored values in 2533 of 4839 recordings (52%). The timing of the recording was also examined in relation to the time at which the recording was solicited; values recorded within 1 hour of the solicited time were judged as acceptable. Of the total of 3342 recordings stored, 2375 (71%) satisfied this criterion. Compliance was significantly less satisfactory in those referred by the Workers' Compensation Board (n = 11). CONCLUSION: In this survey of 21 subjects investigated for possible occupational asthma, compliance with PEF recording, as assessed by comparing recorded and stored results and the time at which the recording was solicited, was poor. PMID- 7499677 TI - Occupational IgE-mediated sensitization and asthma caused by clam and shrimp. AB - BACKGROUND: To confirm occupational asthma caused by clam and shrimp in a food company worker, the following investigation was planned in 60 other exposed workers (56 participants). METHODS: Before the production period of clam and shrimp, a medical and occupational questionnaire was carried out and skin and RAST testing were done with common inhalants and clam, shrimp, crab, and lobster extracts. During the production period, environmental monitoring was performed with personal and general samplers; inhalation testing with methacholine was proposed to subjects with immediate skin reactivity to clam, shrimp, or both. After the production period, all subjects with an immediate skin reactivity to clam, shrimp, or both and either a history of rhinoconjunctivitis, asthma, or bronchial hyperresponsiveness were seen by a specialist. RESULTS: Including the index case in whom occupational asthma to clam and shrimp had been confirmed, four (7%) subjects had a history of rhinoconjunctivitis and two (4%) had a history of asthma during the period of clam production, whereas three (5%) subjects had rhinoconjunctivitis and two (4%) had asthma during the shrimp production. Three (5%) subjects had immediate skin reactivity to clam, and nine (16%) subjects to shrimp. Four (7%) subjects had increased specific IgE antibodies (RAST binding > or = 3%) to clam and eight (14%) to shrimp. Significant associations were found between immunologic reactivity to clam and shrimp on the one hand and to crab and lobster on the other. Environmental monitoring demonstrated clam and shrimp on the air sampling filters. Occupational asthma caused by clam was confirmed by specific inhalation challenges in one subject in addition to the index case. These two subjects had skin reactivity and increased specific IgE antibodies to clam, shrimp, or both. CONCLUSION: By including the initial subject, the prevalence of immediate sensitization is 5% to 7% to clam and 14% to 16% to shrimp. Two (4%) subjects had occupational asthma caused by clam, and one (2%) had occupational asthma caused by shrimp. PMID- 7499680 TI - Characterization of the allergen(s) in latex protein extracts. AB - BACKGROUND: Immediate hypersensitivity to latex, induced by natural latex proteins remaining on the finished products, may lead to severe anaphylactic reactions. METHODS: We investigated the distribution of latex proteins by molecular weight and identified the specific allergenic molecules. Proteins extracted from various latex products were compared with those extracted from raw latex sap, both ammoniated and nonammoniated. RESULTS: Variations in the levels of extractable protein, as well as in the number of molecules and the molecular weight distribution, were observed especially among finished latex products. To identify allergenic (i.e., IgE-binding) molecules, we performed immunoblots with the sera from latex-sensitive persons. The results indicated that antigenic molecule profiles differed among the products and also between the finished products and the raw material. In addition, specificities of the anti-latex IgE antibodies varied among the sensitized persons. CONCLUSIONS: It appeared that persons with the same history of sensitization had similar patterns of antigenic specificities. If the history of exposure, as well as genetic predisposition and medical history of the patient, plays a significant role in the specific IgE response, it may be difficult to select a "standard" antigen and a "standard" antiserum for the evaluation of the latex sensitivity and allergenicity. PMID- 7499679 TI - Defect in potentiation of adenylyl cyclase correlates with bronchial hyperreactivity. AB - BACKGROUND: Adenylyl cyclase is a transmembrane signaling system involved in the inhibition of cellular responses. Recently, we showed that the activity of adenylyl cyclase may be potentiated by stimuli that induce an increase of cellular responses but that do not activate adenylyl cyclase. This is probably an important physiologic feedback mechanism that prevents cells from becoming "overstimulated." OBJECTIVE: Because increased cellular activities are frequently observed in persons with asthma, we hypothesized that a defect in potentiation of adenylyl cyclase might be involved. METHODS: Potentiation of isoprenaline-induced adenosine cyclic monophosphate (cAMP) production with the mitogen phytohemagglutin (PHA; 45 micrograms/ml) or the calcium ionophore A23187 (1 mumol/L) was studied in peripheral blood mononuclear cells taken from patients with asthma (n = 8) and healthy control subjects (n = 11). RESULTS: Isoprenaline induced cAMP production was potentiated significantly in the healthy control subjects (PHA, 110% +/- 15%; A23187, 92% +/- 25%). In contrast, potentiation was not seen with PHA or A23187 in the total group of patients with asthma. However, some patients showed weak potentiation, whereas in others PHA decreased isoprenaline-induced cAMP production. Moreover, the effect of PHA on isoprenaline induced cAMP production correlated significantly with the degree of bronchial hyperreactivity in patients with asthma (r = 0.96; p = 0.0001). CONCLUSIONS: The observed defect in signal transduction could play an important part in bronchial hyperresponsiveness. PMID- 7499681 TI - Diurnal variation of urinary leukotriene E4 and histamine excretion rates in normal subjects and patients with mild-to-moderate asthma. AB - BACKGROUND: Leukotriene E4 (LTE4) and histamine excreted into the urine reflect the in vivo synthesis and release of cysteinyl leukotrienes and histamine, respectively. We examined the diurnal variation of the excretion rate of these mediators over 4 consecutive days in normal subjects (n = 5) and patients with stable mild-to-moderate asthma (n = 8). METHODS: Sixteen consecutive 6-hour urine samples were collected over 4 days. Urinary LTE4 concentrations were determined by reverse-phase high-pressure liquid chromatography, followed by ELISA. Urinary histamine concentrations were measured by ELISA. The excretion rates of these compounds were normalized relative to urinary creatinine content. RESULTS: The mean urinary LTE4 excretion rate was 83.8 +/- 38.2 pg/mg creatinine (mean +/- SD) in normal subjects; in patients with asthma, the urinary LTE4 excretion rate (110.0 +/- 59.2 pg/mg creatinine) was significantly higher than that in normal subjects (p < 0.05). The urinary histamine excretion rate was not different between normal subjects (24.0 +/- 12.5 ng/mg creatinine) and patients with asthma (31.5 +/- 25.8 ng/mg creatinine). A robust and systematic within-day variation (p < 0.01), but no day-to-day variation, was observed in histamine excretion rate. Although the magnitude of variation in LTE4 excretion within a day was significantly greater in patients with asthma than in normal subjects (p < 0.05), we could not identify any specific diurnal variation pattern in either the normal or the asthma group. No significant correlation was observed between urinary LTE4 and histamine excretion rate within any subject. CONCLUSIONS: Patients with asthma excrete LTE4 in the urine at a greater rate than normal subjects. Although no systematic variation in urinary LTE4 excretion rates over the course of a day was observed in either normal subjects or patients with stable asthma, the presence of a systematic diurnal variation of urinary histamine excretion exists in both groups. PMID- 7499678 TI - Constitutive expression of endothelin in bronchial epithelial cells of patients with symptomatic and asymptomatic asthma and modulation by histamine and interleukin-1. AB - BACKGROUND: An upregulation of endothelin-1 expression occurs in bronchial epithelial cells of asthmatic patients. This peptide may mediate bronchoconstriction in asthma, but the mechanisms that modulate endothelin-I synthesis and release are unknown. OBJECTIVE: This study was done to compare the pattern of endothelin-1 expression in patients with symptomatic and asymptomatic asthma and evaluate the ability of inflammatory factors to upregulate endothelin 1 synthesis and release in the epithelial cells of subjects who are free of symptoms. METHODS: Two groups of 10 asthmatic patients were selected. One group had symptomatic asthma with airflow obstruction and moderately to severely increased airway responsiveness. The second group was free of symptoms: they did not show airflow obstruction, and airway responsiveness was borderline or slightly increased. Bronchial biopsy specimens were obtained by means of bronchoscopy and used for immunohistochemical evaluation, epithelial cell isolation, and stimulation experiments with interleukin-1 and histamine. RESULTS: Endothelin-1 immunoreactivity was detected in vivo in the bronchial epithelial cells of all the patients with symptoms and in only two subjects without current symptoms. Incubation of bronchial epithelial cells from patients with asymptomatic asthma with interleukin-1 or histamine, for 8 to 24 hours, resulted in increased expression of endothelin-1 messenger RNA and release of appreciable amounts of the peptide to the culture medium. Those effects were dose- and time dependent. Histamine and interleukin-1 were effective at concentrations similar to those detected in the bronchoalveolar lavage of patients with symptomatic asthma. CONCLUSION: Endothelin-expression is upregulated in bronchial epithelial cells of asthmatic patients with symptoms and evidence of functional derangement as compared with patients without symptoms and airflow obstruction. Exposure of cells from patients with asymptomatic asthma to factors that are released during acute exacerbation of the disease induces endothelin synthesis and release. PMID- 7499682 TI - Association of interleukin-2 and interferon-gamma production by blood mononuclear cells in infancy with parental allergy skin tests and with subsequent development of atopy. AB - The mechanisms regulating the onset of atopic sensitization in human beings are not yet fully clarified. We assessed the capacity of mitogen-stimulated umbilical and peripheral blood mononuclear cells to produce interferon-gamma (IFN-gamma) and interleukin-2 (IL-2) at birth and at 9 months of age in 159 infants. Mononuclear cell production of both IFN-gamma and IL-2 at 9 months, but not at birth, was found to be inversely related to parental immediate skin test reactivity to seven local aeroallergens. Skin test reactivity at the age of 6 years was also inversely related to IFN-gamma and IL-2 production at 9 months of age. However, no relationship was evident between total serum IgE levels at 6 years and production of these cytokines at 9 months. The proportions of circulating lymphocytes and CD4+ or CD8+ cells were also unrelated to skin test reactivity at the age of 6 years. These data suggest that mechanisms regulating skin test reactivity to inhaled allergens may involve deficient IFN-gamma production, deficient IL-2 production, or both during or preceding the time of initial sensitization and that additional mechanisms are involved in regulating total serum IgE level. PMID- 7499683 TI - Eosinophilic inflammation is associated with elevation of interleukin-5 in the airways of patients with spontaneous symptomatic asthma. AB - In vitro and in vivo studies have shown an important role for interleukin-5 (IL 5) in regulating eosinophil proliferation, survival, and effector function. Because eosinophilic inflammation is an important component of symptomatic episodes of asthma, we have investigated whether increased levels of IL-5 protein are present in bronchoalveolar lavage (BAL) fluid of patients with spontaneously symptomatic asthma (FEV1, l61% predicted; FEF25%-75%, 30% predicted) compared with patients with asymptomatic asthma (FEV1, 88% predicted; FEF25%-l75%, 76% predicted). The percent of BAL eosinophils (10.5% vs 0.6]) (p = 0.0001) and eosinophil-derived neurotoxin (386.0 ng/ml vs 6.3 ng/ml) (p = 0.0001) was greater in BAL fluids derived from patients with symptomatic asthma compared with patients with asymptomatic asthma. Levels of IL-5 measured with an immunoradiometric assay were significantly higher in patients with symptomatic asthma (n = 26) compared with those with asymptomatic asthma (n = 18) (274 pg/ml vs < pg/ml) (p = 0.02). The increased IL-5 levels were noted in a subset of patients with symptomatic asthma with BAL absolute eosinophil counts greater than 10(6) (IL-5, 664 pg/ml; n = 10) as opposed to patients with symptomatic asthma with BAL eosinophil counts less than 10(6) (IL-5, < 13 pg/ml; n = 16) (p = 0.005). This study suggests that IL-5 is not only induced in experimental models of allergen-induced asthma but can also be detected as asthma progresses from the asymptomatic to the clinically symptomatic state in subjects with significant BAL eosinophilia. PMID- 7499685 TI - One of the rubber latex allergens is a lysozyme. AB - BACKGROUND: Type I hypersensitivity reactions caused by latex products are ascribed to proteins eluted from them, but little is known about the properties of these allergenic proteins. The reason for the cross-reaction between rubber latex and fruits is also not known. We have speculated that a series of defense related proteins in plants is a cause of latex allergy and the cross reaction. OBJECTIVE: To verify our hypothesis, we selected a lysozyme as a representative defense-related protein and examined its relationship to latex allergy. METHODS: Lysozymes eluted from latex gloves were detected with a cell-suspension clearing test. A chromatographically separated lysozyme was investigated for its physicochemical and enzymatic properties and allergenicity. RESULTS: Lysozyme activity was detected in extracts from ammoniated latex and latex gloves. We separated a lysozyme (27 kd; isoelectric point, 9.5) using cation-exchange and gel filtration chromatography. This lysozyme was enzymatically very similar to fruit lysozymes and was demonstrated to be an allergen. CONCLUSIONS: One of the rubber latex allergens is a lysozyme that has similarities to fruit lysozymes. This suggests the relevance of defense-related proteins to latex allergy and the cross reaction. PMID- 7499684 TI - Effect of six-hour exposure to nitrogen dioxide on early-phase nasal response to allergen challenge in patients with a history of seasonal allergic rhinitis. AB - BACKGROUND: Recent studies have suggested that exposure to air pollutants may enhance the airway responsiveness of susceptible individuals to inhaled allergen. METHODS: To investigate the effect of exposure to nitrogen dioxide (NO2) on nasal airways resistance (NAR) and inflammatory mediators in nasal lavage fluid, eight subjects with a history of seasonal allergic rhinitis, who were tested out of season, were exposed in a randomized single-blind, crossover study to either air or 400 ppb NO2 for 6 hours. The changes in NAR and eosinophil cationic protein (ECP), mast cell tryptase (MCT), neutrophil myeloperoxidase (MPO), and interleukin-8 (IL-8) in nasal lavage fluid before and after exposure were evaluated. Another group of eight subjects with a history of seasonal allergic rhinitis were also randomized to exposure to air or 400 ppb NO2 for 6 hours and then challenged with allergen, before evaluation for changes in NAR and changes in ECP, MCT, MPO, and IL-8 in nasal lavage fluid. RESULTS: Exposure to air or NO2 did not alter either NAR or the levels of ECP, MCT, MPO, or IL-8 in nasal lavage fluid. Allergen challenge after exposure to both air and NO2 significantly (p < 0.05) increased levels of MCT, but not MPO and IL-8 in the nasal lavage fluid. In addition, allergen challenge after exposure to NO2 but not air, significantly increased levels of only ECP in nasal lavage fluid (p < 0.05). CONCLUSIONS: These results suggest that acute exposure to NO2 at concentrations found at the curbside in heavy traffic during episodes of pollution, may "prime" eosinophils for subsequent activation by allergen in individuals with a history of seasonal allergic rhinitis. PMID- 7499686 TI - Antidopaminergic antibodies in a patient with a complex autoimmune disorder and rapidly progressing Parkinson's disease. PMID- 7499687 TI - Allium cepa seeds: a new occupational allergen. PMID- 7499688 TI - Occupational asthma caused by grass juice. PMID- 7499689 TI - IgE binding to a human profilin-containing fraction caused by contamination with yeast-derived material. PMID- 7499690 TI - Noncardiogenic pulmonary edema induced by nonionic low-osmolality radiographic contrast media. PMID- 7499692 TI - Insect sting challenges: do no harm. PMID- 7499691 TI - Prevalence of latex allergy among greenhouse workers. PMID- 7499694 TI - Analysis of insect-sting study. PMID- 7499693 TI - Sting challenges as criteria for venom immunotherapy. PMID- 7499695 TI - Insect-sting challenges--all risk and no benefit? PMID- 7499696 TI - Sting challenges. PMID- 7499697 TI - Serum eosinophilic cationic protein as a marker of disease activity in chronic asthma. PMID- 7499698 TI - Parental care and women's health. PMID- 7499699 TI - Not all prenatal care is appropriate care. PMID- 7499700 TI - Prenatal care--the good, the bad, and the ugly. PMID- 7499701 TI - Family planning: an essential component of prenatal care. AB - This nation's traditional approach to improving maternal and infant health has been prenatal care. But evidence is mounting that additional progress in reducing maternal and infant morbidity and mortality will depend, at least in part, on the care that a woman receives before she conceives. The studies reviewed in this paper indicate that increasing the interval between deliveries and preventing or delaying pregnancies among women at high risk could lower the rate of low birthweight (LBW). Since reducing the rate of unintended pregnancies would also reduce the number of pregnancies in women at high risk of LBW because of race, age, late or no prenatal care, and unhealthy behaviors, the prevention of unintended pregnancies would also reduce LBW. Unfortunately, prenatal care, as experienced by many women, devotes little attention to these family planning issues. Many women do not realize the importance of family planning to their own health and that of their children. Prenatal care providers should include instruction about the importance of pregnancy planning and encourage women to continue receiving health care between pregnancies. If the health of women and infants is to be improved, society must be willing to provide health services to women of reproductive age even when they are not pregnant. PMID- 7499702 TI - The importance of extreme prematurity and low birthweight to US neonatal mortality patterns: implications for prenatal care and women's health. AB - OBJECTIVE: In order to frame the appropriateness of neonatal mortality reduction efforts that begin only after pregnancy is recognized, this study examined the relative contributions of different gestational age and birthweight groups to total neonatal mortality and to racial disparities in neonatal mortality in the United States. METHODS: Using the national linked birth/infant death data set for the 1988 cohort, the relative contributions of different birthweight and gestational age groups to national neonatal mortality rates were calculated. The relative contributions of these groups to the racial disparity in neonatal mortality were also assessed. RESULTS: Very low birthweight infants (< 1,500 g) accounted for 1.2% of all births, but 64.2% of all neonatal deaths. The very low birthweight rate for whites was 0.93%, while that for blacks was 2.79% with the contribution of this group to neonatal mortality higher for blacks than whites. Infants less than 1,000 g contributed more than 80% of the racial disparity in neonatal mortality. CONCLUSION: Neonatal mortality patterns in the United States have become highly dependent on infants with gestational ages that approach the second trimester. Preventing neonatal mortality by enhancing care only after pregnancy has been recognized, therefore, may be limited. Strategies that link prenatal care to broader initiatives to improve the health of women regardless of pregnancy status may be more effective. PMID- 7499703 TI - Prenatal care and public policy: lessons for promoting women's health. AB - One of the most notable health-related legislative achievements of the 1980s was the expansion that transformed Medicaid from a medical assistance program for a relatively small group of the very poorest mothers meeting strict categorical criteria to a health program for low-income uninsured, pregnant women and their children, regardless of work status or family composition. Yet Medicaid coverage remains unavailable to millions of poor and uninsured women who are significantly less likely to obtain needed care without health insurance. Among the factors that help explain why Medicaid was not simultaneously expanded for uninsured women are our nation's longstanding reluctance to provide public assistance except to individuals deemed "deserving" based on the circumstances that led to their indigency. Another factor is the special value our culture places on pregnancy, which permits pregnant women who do not meet the definitions of the "deserving" to transcend those restrictions. Finally, while data served as an effective tool in the campaign for pregnant women's coverage, comparable data related to nonpregnant women are limited. Nonetheless, the lessons from the Medicaid expansion are helpful in efforts to improve women's access to care in general. PMID- 7499704 TI - Missed opportunities for intervening in the lives of pregnant women addicted to alcohol or other drugs. AB - This study examined the practices of health care providers in detecting substance abuse among pregnant women and making appropriate referrals for treatment and other services. Data were gathered through personal interviews with 181 addicted women during visits to four sites and through a survey of 94 substance abuse treatment programs in the South. The typical woman in the sample was 31 years old, white, divorced or separated, and had been using alcohol and other drugs for 12.7 years. A majority (65%) had been asked about alcohol and drug use by physicians or nurses during their most recent pregnancies. Of those who were asked, 43% admitted alcohol or drug use, and an additional 13% were using but did not admit it. Very few women (5%) were referred to drug counselors or programs. A majority (59%) were not given information about the effects of alcohol or drug use on pregnancy. Even when prenatal care providers ask addicted women about substance abuse, they often fail to make appropriate referrals to treatment. PMID- 7499706 TI - Employment-related physical activity and pregnancy outcome. AB - Concern about possible adverse reproductive effects of employment reflects the increasing participation of pregnant women in the labor force. Although the literature on the effect of employment-related physical activity on pregnancy outcome is substantial, no general consensus has been reached. A growing number of studies, however, suggests that prolonged standing and long working hours may pose a risk to pregnancy. The provision of maternity benefits varies greatly between the United States and European countries. Current modifications of working conditions for both pregnant and non-pregnant women differ even among European countries. PMID- 7499705 TI - Efforts to reduce perinatal mortality, HIV, and drug addiction: surveys of the states. PMID- 7499707 TI - Making a difference for children: the medical community's role in improving child care. AB - The quality of child care for the majority of children in the United States is poor to mediocre. The lack of good quality child care has serious consequences for family stability, work force productivity, and child health and development. The medical community, especially pediatricians, family practitioners, nurses, and nurse practitioners, can play five key roles in helping to ensure that every child has a solid early foundation: 1) they can become trusted sources of support for parents, who are often isolated from their extended families and unsure about their new roles as parents; 2) they can give child care providers in the community information about health and safety practices; 3) they can provide preventive and diagnostic services to children in child care programs; 4) they can act as advocates for improved services to children and families; and 5) they can refer parents and siblings for care. This paper describes some of these efforts and calls for their replication in communities throughout the country. It also describes policies in other countries that have improved children's health and development and reduced child poverty and its related impact on health. PMID- 7499708 TI - Heterogeneity between women who received prenatal care in the third trimester and those who received no prenatal care. AB - OBJECTIVE: To determine whether women who received prenatal care in the third trimester differed from those who received no prenatal care. METHODS: We analyzed US birth certificates from 1990 through 1992, computing the distribution of live births for women who received prenatal care in the third trimester and for those who received no prenatal care according to eight demographic and pregnancy related characteristics (age, race, marital status, residence, country of birth, education, interbirth interval, and parity). We used the Cochran-Mantel-Haenszel statistic to test the significance of the differences between the distributions for each characteristic, adjusting simultaneously for the other seven characteristics. RESULTS: Women who received no prenatal care differed from women who received prenatal care in the third trimester for each of the demographic and pregnancy-related characteristics we examined. Among black and unmarried women, the two categories of prenatal care differed by more than 10%. CONCLUSIONS: The characteristics of women who received no prenatal care and those of women who received prenatal care in the third trimester were heterogeneous. Strategies to promote earlier prenatal care should be specific and sensitive to women at risk for each category of late entry to prenatal care. PMID- 7499709 TI - Baby love and budget relief: some promising practices in prenatal managed care in Medicaid. AB - The provision of prenatal health care is one of the most important functions of Medicaid, as approximately three-quarters of program enrollees are poor women and children. A rapidly increasing number of states are seeking to address the dual crises of spiralling program costs and inadequate access to services through the use of managed care in Medicaid. Medicaid managed care is growing rapidly and now stands at more than 8 million enrollees, the overwhelming majority of whom are women and children. The focus on accessible preventive and primary care that is, in theory, the hallmark of managed care, should apply well to prenatal health services for Medicaid beneficiaries, though this is not always the case. The more successful managed prenatal care programs thus far have some common characteristics, which are examined here in the hope that they will be replicated by other states. PMID- 7499710 TI - Prenatal care in developing counties: the World Health Organization technical working group on antenatal care. AB - As part of its work to help define and strengthen prenatal care, the World Health Organization convened a Working Group in November 1994 to formulate recommendations for prenatal care at the health center level. The Group agreed on the timing and content of a minimal number of visits for all women, as well as risk factors and medical conditions that should receive special care. In addition, several over-riding themes came out of the discussions. These included the need for prenatal care to be tailored to each woman's circumstances, the importance of developing a delivery plan appropriate to the woman's situation, the need to make prenatal care part of a functioning health care system in order for it to be effective, the appropriateness of the midwife as provider of prenatal care, and the need to educate and empower the entire community to address the causes of maternal morbidity and mortality. Operational research and evaluation of prenatal care practices are underway to identify the most effective and efficient ways to provide these services. PMID- 7499711 TI - Framing the debate: can prenatal care help to reduce the black-white disparity in infant mortality? AB - Prenatal care has been identified as necessary to reducing the disparity in infant mortality between black and white infants. The purpose of this paper is to review determinants of the disparity and describe the contribution that prenatal care can make to modifying those determinants, biologic or social. When examined by birthweight categories, 25% of the disparity is due to excess deaths among normal birthweight infants (> or = 2500 g); 13% is due to excess deaths among moderate low birthweight infants (1500-2499g); and 62% of the disparity is due to excess deaths among very low birthweight infants. Normal birthweight black infants have higher rates of death due to infections, injuries, and sudden infant death syndrome. Very low birthweight black infants account for the increasing disparity in infant mortality. Social determinants of the disparity in infant mortality include the effects of poverty and the accompanying problems of limited access to health care services, preventive care, and good nutrition. Prenatal care may reduce the disparity by using both high-risk and population-based prevention strategies. This combination of strategies would identify and treat medically high-risk women before delivery and provide preventive care to all women, regardless of their risk status. Although both strategies have a potential for producing modest reductions, neither has proved to be effective. PMID- 7499712 TI - Driving vision. PMID- 7499713 TI - The effect of redeepening incisions on radial keratotomy outcome. AB - BACKGROUND: Patients scheduled to undergo radial keratotomy in both eyes received redeepening incisions in one eye and single pass incisions in the other. Eye and order of surgery were assigned randomly and surgical parameters were otherwise identical in the two eyes. METHODS: This study was designed to measured the improvements in myopia obtained by radial keratotomy, with and without redeepening incisions. RESULTS: There was a greater decrease in spherical equivalent manifest refraction in the redeepening group than in the control group at 1 week (0.41 D) and 1 month (0.36 D) after surgery, as measured by keratometry. The difference in change between groups was 0.41 D at one week, 0.36 D at 1 month, and 0.26 D at 3 months. Change in spherical equivalent was significantly more in the redeepened group at 3 months after surgery (0.37 D); however, the improvement constituted only 8.8 percent of the decrease in myopia achieved by radial keratotomy without redeepening (3.87 D). CONCLUSIONS: Although redeepening yields an incremental effect in the correction of myopia, the technical difficulties of redeepening and the increased chance of perforation of the cornea with redeepening outweigh the benefits of the procedure. PMID- 7499714 TI - Results of a vision screening program: an initial study. AB - BACKGROUND: More than 100 children between the ages of 5 and 13 years (grades K through 6) were screened with the New York State Optometric Association vision screening battery. METHODS: A comparison between regular and special education referral rates was performed. Two groups of tests were established: acuity testing and functional testing. A comparison of the failure rates of these two groups was also analyzed. RESULTS: This study revealed that although the referral rate between the two populations showed no statistical difference, the particular "group" of tests failed differed significantly. CONCLUSIONS: Although no conclusions can be drawn from these numbers as the sample was not varied enough, further screenings in different geographical and socioeconomic areas need to be performed to determine the validity of this trend. PMID- 7499715 TI - Comparison of locus of control among nursing home residents with visual impairment and normal sight. AB - BACKGROUND: Despite well-documented effects of psychosocial factors on patients with visual impairment, eye care professionals often fail to incorporate these important concepts into management plans for this population. METHODS: This study reports findings using the Levenson Locus of Control (LoC) Scale and its two modifications by Shewchuk et al. The study compares nursing home residents with visual impairment to individuals having normal sight to determine the status of LoC as a factor in visual rehabilitation. RESULTS: T values vary significantly in LoC scores of nursing home residents sampled when compared with variables of gender and type of residence (i.e., independent living, congregate living, assisted living, intermediate care). T values do not vary significantly, however, across variables of visual impairment and normal sight. CONCLUSIONS: LoC does not appear to be a psychosocial factor that varies significantly among elderly nursing home residents sampled across the characteristics of normal sight and visual impairment. However, the population sampled may possess qualities that prevent generalization to the elderly population as a whole. A need for research with a larger, more diverse sample using locus of control scales as well as research using other psychosocial tests to evaluate patients with visual impairment is indicated. PMID- 7499716 TI - Second opinion: a computer expert system designed to aid in the determination of spectacle lens prescriptions. AB - BACKGROUND: Expert systems (ESs) are computer programs that simulate the decision making ability of human experts. In this project, an ES was developed that used optometric data and decision rules to determine spectacle lens prescription powers. METHODS: Decision rules derived from clinical experience and standard references were provided to a commercially available shell program to create an ES called Second Opinion. The ES determined spherical, cylindrical, and add powers designed to provide maximum acuity consistent with a high probability of patient acceptance. CONCLUSIONS: Second Opinion demonstrates how a computer program can be used to analyze optometric data and offer suggestions regarding lens prescriptions. PMID- 7499717 TI - Community-oriented primary care programs. AB - BACKGROUND: Systems are being proposed by health planners in an effort to improve access to health care for the uninsured and underinsured, including the vulnerable and indigent populations. The lack of affordable and adequate health care for significant numbers of our citizens raises the potential for a serious economic crisis for our nation. Modern medicine has been more interested in developing high tech care for the well insured than in preventive and early intervention health care. While the development of this high technology is of great value and will be beneficial to many, it is out of financial reach for increasing numbers of individuals. As a nation we must reduce the demand for high priced medical care by providing a system that keeps people healthy or intervenes in their illness at a stage when less dramatic medical measures are able to restore them to health. To maintain a population that is healthy and productive in the school setting and in the work place, systems must be developed that reach out into the community with prevention-oriented health education programs and health care services at the primary care level. METHODS: A limited literature review was made in order to present to the optometric community the concept of Community Oriented Primary Care (COPC) as a cost-effective method of improving access to primary care services. RESULTS: The literature review demonstrated that Community Oriented Primary Care (COPC) as a health-oriented strategy brings the benefits of primary care closer to those in need. The history of the development of community oriented primary care services as a health delivery methodology, as well as the current utilization of the COPC concept, is presented in this paper. CONCLUSIONS: Optometry, as a provider of primary eye and vision care services should be included in these cost effective programs. PMID- 7499718 TI - Vision profile of patients with mild brain injury. AB - BACKGROUND: Patients suffering from a traumatic brain injury (TBI) show many types of visual dysfunctions, including binocular, oculomotor, accommodative and visual field loss. This study evaluates the visual status of patients suffering a mild TBI. METHODS: The visual findings of a control group of 16 non-TBI patients are compared to the visual findings of 16 patients with documented mild TBI. Functions compared include symptomology, refractive status, stereopsis, ocular alignment by cover test, relative accommodation, pursuits, near point of convergence break and recovery, phorias at distance and near and vergence ranges at distance and near. RESULTS: Significant differences were found between the two groups in the areas of refractive status, near point of convergence break/recovery, near cover test, stereo acuity and base-in break at distance. There were also significant differences in symptomology, pursuit function, base in recovery at distance, base-in break/recovery at near and base-out recovery at distance. CONCLUSIONS: In-depth visual evaluation of mild TBI patients is critical for documentation purposes as well as for possible treatment regimens. PMID- 7499719 TI - Circumscribed choroidal hemangioma. AB - BACKGROUND: Circumscribed or solitary choroidal hemangioma is a relatively rare, benign vascular tumor. Because its appearance may mimic more serious intraocular lesions, eye care practitioners should be familiar with differential diagnosis and treatment options. METHODS: A patient presented with an asymptomatic, elevated, red-orange lesion above the macula in the right eye and a history of longstanding unexplained visual acuity loss in the eye. Intraocular lesions such as amelanotic melanoma, metastatic tumor, choroidal osteoma, disciform scar, serous detachment, and central serous chorioretinopathy can appear similar to circumscribed choroidal hemangioma (CCH) on ophthalmoscopy, but may be differentially diagnosed with fluorescein angiography, ultrasonography, and periodic observation. RESULTS: CCH requires various therapeutic options. Until the patient becomes symptomatic from visual loss secondary to serous macular detachment or subretinal neovascular membrane formation, no treatment is necessary. Laser photocoagulation is the preferred therapeutic intervention, and most patients will require a minimum of two sessions. A small percentage of individuals will progress to complete retinal detachment with profound visual loss regardless of therapy. Most CCHs remain stable in size and do not undergo malignant transformation. CONCLUSIONS: CCHs may be overlooked because of their initial subtle clinical appearance with no associated cutaneous or systemic anomaly. Even though CCH is a benign tumor, it may mimic more serious intraocular lesions and require a combination of ancillary tests for proper diagnosis. Depending on visual symptoms or potential threat to vision, management options include periodic observation, laser photocoagulation, cryotherapy, microwave therapy, and possible enucleation. CCH has little or no growth potential, but visual prognosis remains guarded due to significant secondary retinal changes that can occur over the tumor's surface. CCH must be considered in the differential diagnosis of an elevated intraocular lesion. PMID- 7499720 TI - Sarcoid optic neuropathy: a case report. AB - BACKGROUND: Sarcoidosis is frequently overlooked as a cause of optic neuropathy. A cascade of immune interactions has been cited in the pathogenesis. Optic nerve involvement can be the initial manifestation of systemic disease, making the diagnosis even more elusive. Compatible clinicoradiographic and abnormal laboratory findings along with histologic evidence of noncaseating granuloma are necessary to secure the diagnosis of sarcoidosis. METHODS: A case report is presented of a 41-year-old patient with severe papillitis and a junctional scotoma as the first declaration heralding neurosarcoidosis (NS). Co-management with a neuroophthalmologist at an academic teaching hospital prompted the necessary diagnostic studies to preserve vision. RESULTS: Sarcoidosis was diagnosed on the basis of the patient's clinical presentation of rapid vision loss, enlarging mass in the anterior visual pathway revealed by CT and MRI, as well as increased uptake of gallium in the lungs and lacrimal glands. Also considered in the differential diagnosis were glioblastoma, lymphoma, and demyelinating disease. CONCLUSIONS: This case represents an example of triage referral involving optometry and academic ophthalmology. Rapid intervention with intravenous corticosteroids should be considered in the face of atypical optic neuritis with a suspected inflammatory etiology. PMID- 7499721 TI - Svyatoslav Fyodorov, M.D.: innovative eye surgeon. AB - BACKGROUND: Svyatoslav Fyodorov is a brilliant, innovative Russian ophthalmic surgeon. METHODS: who has sought to restore eyesight to as many people as possible. His unconventional methods have surprised, shocked, and antagonized his medical colleagues and have challenged the accepted therapies of the day. He has nevertheless persevered and has ultimately won acceptance and respect. RESULTS: In an effort to uncover the workings of the eye, he investigated the surgical effort to substitute an artificial intraocular lens for a cataractous one, which challenged the accepted techniques of the time. He did this by the use of radial corneal incisions to eliminate myopia, which was performed on a conveyor belt. CONCLUSIONS: Fyodorov's surgical technique for the reduction of myopia, radial keratotomy, has been modified and performed on hundreds of thousands of eyes in the United States. PMID- 7499722 TI - Effects of hypercortisolemia or hyperinsulinemia on neurochemical indices of catecholamine release and synthesis in conscious rats. AB - Glucocorticoids and insulin (INS) complexly affect sympathoneural and adrenomedullary outflows. This study assessed effects of chronic hypercortisolemia and effects of INS independent of INS-induced hypoglycemia on neurochemical indices of different aspects of catecholaminergic function in conscious rats. Since L-DOPA is the precursor of the endogenous catecholamines and the immediate product of the rate-limiting enzymatic step in catecholamine biosynthesis, alterations in rates of appearance (spillover) of L-DOPA in arterial plasma may reflect alterations in catecholamine synthesis. The study therefore included examination of whether cortisol (CORT) or INS affects L-DOPA spillover or renal excretion of dopamine (DA) derived from plasma L-DOPA. Arterial plasma levels and urinary excretion rates of endogenous catechols and radiolabelled L-DOPA and DA were measured during systemic intravenous infusions of [3H]L-DOPA. CORT was administered via a subcutaneous minipump reservoir for one week prior to [3H]L-DOPA infusion, and INS was infused with glucose to examine effects of hyperinsulinemia independently of hypoglycemia. CORT decreased plasma levels and urinary excretion of norepinephrine (NE). INS did not. Neither CORT nor INS affected levels of other catechols, L-DOPA spillover, or the rate of urinary excretion of [3H]DA for a given plasma level of [3H]L-DOPA. The results suggest that CORT inhibits sympathetically-mediated NE release without altering overall rates of catecholamine turnover or synthesis in sympathetic nerves in vivo and that INS effects on catecholaminergic function depend entirely on INS induced hypoglycemia. PMID- 7499723 TI - Distribution of noradrenergic neurons in the female rat pelvic plexus and involvement in the genital tract innervation. AB - The involvement of the pelvic plexus noradrenergic neurons in the innervation of the genital tract was studied in the female rat. Several small ganglia were observed in addition to the paracervical ganglion and immunocytochemistry for tyrosine hydroxylase was performed to examine the distribution and number of the noradrenergic neurons. 5069 +/- 1525 nerve cell bodies were counted in the paracervical ganglion and 9.0 +/- 0.8% of them were noradrenergic, displaying a clear somatotopic distribution in the ventro-medial part of the ganglion. Some accessory ganglia were located ventral to the main paracervical ganglion. 414 +/- 149 nerve cell bodies were found in the accessory ganglia, of which 20.4 +/- 3.1% were noradrenergic. Ganglia along the vesical branch of the hypogastric nerve, referred to as an hypogastric plexus, contained 233 +/- 83 neurons among which 12.7 +/- 7.2% were noradrenergic. Bilateral removal of the pelvic plexus produced degeneration of all the tyrosine hydroxylase-immunoreactive nerve fibres in the lower part of the uterus and in the cervix. In contrast, excision of the paracervical ganglia and the accessory ganglia caused no significant change in this innervation pattern. Combined retrograde tracing study and immunocytochemistry for tyrosine hydroxylase revealed a very small number of noradrenergic neurons also labelled with fluoro-Gold. Both findings suggest a limited involvement of the pelvic plexus noradrenergic neurons in the innervation of the lower genital tract. PMID- 7499724 TI - Location of sensory nerve cells that provide calbindin-containing laminar nerve endings in myenteric ganglia of the rat esophagus. AB - To determine the origin of the calbindin-containing laminar nerve endings in the myenteric ganglia of the rat esophagus, retrograde tracing experiments combined with immunohistochemistry using an antibody for calbindin were carried out. After Fast blue was injected into the cervical portion of the esophagus, labeled neurons were found bilaterally in the nodose ganglion and dorsal root ganglia of C1 to T3. 80% of the total neurons in the nodose ganglion and 20% of those in the dorsal root ganglia showed calbindin immunoreactivity. Moreover, 79% of Fast-blue labeled neurons found in the nodose ganglion and 18% of those in the dorsal root ganglia were immunoreactive for calbindin. These results suggest that the calbindin antibody we used is useful as a marker for identifying esophageal vagal afferents derived from the nodose ganglion. The calbindin-immunoreactive nerve fibers forming the laminar endings in the myenteric ganglia of the rat cervical esophagus are mainly derived from sensory neurons in the nodose ganglion and partly derived from those in the cervical and upper thoracic dorsal root ganglia. Calbindin-containing laminar nerve endings may be related to mechanoreceptors in the esophagus. PMID- 7499725 TI - The effect of interleukin-1 beta on the efferent activity of the vagus nerve to the thymus. AB - The effects of intravenous (i.v.) administration of recombinant human interleukin 1 beta (rhIL-1 beta) and human interleukin-1 beta-fraction (hfrIL-1 beta) on the activity of vagal efferent fibers innervating the thymus were observed in urethane-anesthetized rats. An i.v. injection of 10 ng rhIL-1 beta or hfrIL-1 beta caused a similar degree of increase in the efferent activity, which lasted longer than 2 h. The least effective dose was 1 ng and dose-dependent responses were observed after i.v. injection of rhIL-1 beta at doses of 1 ng, 10 ng and 100 ng. The IL-1 beta-induced activation of efferent activity of the vagal branch to the thymus implicates their involvement in the neural modulation of the thymic function. PMID- 7499726 TI - Does low frequency power of arterial blood pressure reflect sympathetic tone? AB - We tested whether power spectral analysis of arterial blood pressure (ABP) is a feasible tool to detect differences in peripheral sympathetic nerve activity in normotensive and hypertensive rats with differing basal sympathetic tones. Nine Wistar Kyoto rats (WKY), 10 Sprague-Dawley rats (SD), 10 spontaneously hypertensive rats (SHR) and 9 hypertensive transgenic rats harbouring the mouse Ren-2 gene (TGR) were chronically instrumented with femoral artery catheters and nerve electrodes around the sympathetic major splanchnic nerve. Two days after surgery ABP and splanchnic nerve activity (SpNA) were recorded in the conscious state during basal conditions as well as during alpha 1-adrenergic receptor blockade. Power spectra and squared coherence in the low (LF, 0.02-0.20 Hz), mid (MF, 0.20-0.80 Hz) and high (HF, respiration peak +/- 0.3 Hz) frequency bands were calculated for ABP and SpNA. Mean blood pressure in SHR (133 +/- 8 mmHg) and TGR (142 +/- 8 mmHg) was significantly higher (P < 0.05) than in WKY (115 +/- 3 mmHg) and SD (95 +/- 4 mmHg). SpNA in SHR was higher than in WKY (23.4 +/- 6.4 microV vs. 11.6 +/- 0.8 microV, P < 0.05) while SpNA in TGR was lower than in SD (20.1 +/- 3.9 microV vs. 28.8 +/- 4.2 microV, P < 0.05). LF and MF components of ABP variability were not significantly higher in those rats with high sympathetic tones. However, alpha 1-adrenergic receptor blockade reduced LF and MF components of ABP and SpNA in all strains except SHR. LF and MF coherence was not greater in rats with high sympathetic tones than in those with low sympathetic tones. The reduction of LF and MF components of ABP variability by alpha 1-adrenergic receptor blockade indicates an important contribution of peripheral sympathetic nerve activity to LF and MF blood pressure variability on an acute basis. However, the lack of higher LF and MF power in the ABP spectra of those rats with high SpNA together with the finding that LF and MF coherence was not higher in those rats with high SpNA led to the conclusion that LF and MF spectral components of ABP do not appear to be suitable markers for the prevailing sympathetic nerve activity. PMID- 7499729 TI - Increased muscle sympathetic nerve activity after glucagon administration in man. AB - Muscle sympathetic nerve activity (MSNA) after glucagon administration was quantitatively measured in five healthy male volunteers (age: 20-28 years old). After an overnight fasting, 1 mg glucagon dissolved in a 1-ml vehicle was injected i.v. as a bolus. MSNA, ECG and blood pressure were simultaneously recorded 15 min before and after the injection. MSNA (bursts/min), heart rate (HR), and systolic and diastolic blood pressures (dBP and sBP) were significantly increased at 2-3 min after glucagon injection. Their means and standard deviations of control for 15 min vs. 3 min after the injection were in the five subjects; MSNA, 10 +/- 6 vs. 30 +/- 10 bursts/min, HR, 61 +/- 10 vs. 76 +/- 8 beats/min, dBP, 66 +/- 9 vs. 91 +/- 9 mmHg and sBP, 122 +/- 12 vs. 152 +/- 11 mmHg. These results suggest that glucagon has a direct facilitatory effect on the central sympathetic nervous system. PMID- 7499728 TI - Haemodynamic responses in muscle and adipose tissue and whole body metabolic responses during norepinephrine infusions in man. AB - Circulatory metabolic and cardiovascular responses to 1-h-long infusions of norepinephrine (NE) (approx. 0.2 and 0.4 nmol/kg body weight per min) were measured on two separate occasions in six subjects. The infusions increased circulating NE concentrations 6- and 13-fold, respectively. Blood flow to adipose tissue, measured with the 133Xe clearance technique, increased from a basal value of about 3 ml/100 g per min, to about twice this value at 60 min with both doses of NE. In contrast muscle blood flow was unaffected. The higher dose of NE produced significant increments at 60 min in whole body oxygen consumption (approx. 9%), and circulating concentrations of glucose (approx. 18%), non esterified fatty acids (approx. 200%) and glycerol (approx. 32%) which were greater than those observed with the low-dose infusion. Changes in blood pressure, pulse and CO2 exchange were observed within 5-10 min after the start of the infusion, whilst changes in adipose tissue blood flow were observed after 15 30 min. It is concluded that in humans (i) a dose of NE as low as 0.2 nmol/kg per min is sufficient to evoke both circulatory and metabolic responses; (ii) the pattern in the adipose tissue blood flow response to NE may help explain some of the conflicting reports about the haemodynamic effects of this hormone in adipose tissue; and (iii) blood flow and vascular resistance in different tissues may be affected in different ways by norepinephrine. PMID- 7499727 TI - Young borderline hypertensives are hyperreactive to mental arithmetic stress: spectral analysis of R-R intervals. AB - To investigate whether sympathetic tone and its reactivity to stress are increased in borderline hypertension, we compared pressor and autonomic nervous responses to mental arithmetic stress in male borderline hypertensives (BH) and normotensive volunteers (NT). Three age groups, 30, 40 and 50-year-old groups, which included 30 to 39, 40 to 49 and 50 to 59-year-old subjects, were studied. Fractional LF (%LF), fractional HF (%HF) and L/H, obtained from the power spectrum of R-R intervals, were used as indices of autonomic nervous function. Baseline autonomic nervous indices did not differ between NT and BH of any age group. Blood pressure rose higher during mental arithmetic stress in 50 than in 30-year-old NT but not in comparable age groups of BH. Pressor responses were augmented in BH compared to NT only in the 30-year-old group. However, the differences were not significant when pressor responses were expressed as percent increases in blood pressure. Both %L and L/H increased during arithmetic stress. The increase in %L did not differ between NT and BH but that in L/H (% delta L/H) was larger in 50 than in 30-year-old NT. % delta L/H was larger in BH than in NT only in the 30-year-old group. These findings suggest that both pressor and autonomic nervous responses to metal arithmetic stress were altered by aging and augmented in BH compared to NT in the 30-year-old group. PMID- 7499731 TI - [Biological surveillance of hormone substitution: relying on the plasma levels of estradiol or the products used?]. AB - The success of hormone substitution in menopaused women is widely recognized. Generally an oestrogen or a progesterone is used. The aim of this work was to determine whether the oestrogen treatment can be monitored on the basis of the induced plasma levels of oestradiol and whether there is any difference between specific products available. A review of the literature and personal observations concerning the pharmacokinetics of different oestrogen therapies has shown that the single assay of oestradiol is insufficient to adapt dosages and that different products are dosed differently giving variable reproductibility between the different formulations available. PMID- 7499730 TI - The spinally mediated 10-Hz rhythm in the sympathetic nerve activity of cats. AB - To examine the origin of the so-called '10-Hz rhythm' in the sympathetic nerves, the mass discharges of the white ramus of the third thoracic segment (T3WR) and the inferior cardiac nerve (ICN) and the activities of single postganglionic neurons in the stellate ganglion were recorded in spinal cats. During the chemical or electrical stimulation of the spinal cord, the time of peak of discharges in the sympathetic nerves was analyzed. Both intrathecal administration of N-methyl-D-aspartic acid (NMDA; 3-10 mM) and continuous high frequency (80-200 Hz) electrical stimulation of the dorsolateral funiculus at the second cervical level increased activity of the sympathetic nerves in a similar fashion. In these conditions, modes of the inter-peak interval histograms (IPIH) were about 100 ms (range; 90-130 ms), the inverse of about 10 Hz, but no correlation was observed in autocorrelograms of these peaks of discharges. Therefore, this 100-ms interval activity might have some significance for the 10 Hz rhythm. In order to make this point clear, we stimulated the dorsolateral funiculus with intermittent trains of electrical pulses (0.2-ms duration, 10-35 pulses of 80-200 Hz frequency, in every 300-800 ms). While intermittent trains of pulses were applied, multiple peaks of discharges were evoked in the sympathetic nerves. IPIHs of the nerves were multimodal. The first mode (shortest interval) was about 100 ms. The first mode depended on none of the stimulus parameters but the probability of the about 100-ms interval activity depended on the interval of trains of pulses and the stimulus intensity. With this intermittent stimulation, the autocorrelogram of the peaks revealed the 100-ms interval rhythm. To confirm that the peak of discharges in the ICN was composed of synchronized spikes of postganglionic fibers, single neuronal activities of postganglionic neurons were recorded during the intermittent stimulation. Inter-spike interval histograms showed almost same profile as the IPIHs of the ICN. These results can be explained if the following two assumptions are valid; (i) There are mechanisms that limit minimum firing interval of most preganglionic neurons to about 100 ms. (ii) Simultaneously a interneuron in the spinal cord resets the spike generation of multiple preganglionic neurons. Similarity of the spike activities of the sympatho-excitatory reticulospinal neurons to the intermittent stimulation can explain the 10-Hz rhythm in the peripheral sympathetic nerves in intact spinal cord animals. It is not necessary to postulate the specific 10-Hz rhythm generator in the brain stem for the sympathetic nervous system. PMID- 7499732 TI - [Intrauterine ultrasonography: first symptomatologic aspects]. AB - A mechanical ultrasonographic probe (7.5 MHz ou 10 MHz was enclosed in a 3.5 mm teflon catheter for intrauterine explorations. Correlative illustrations of normal and pathological images are presented. PMID- 7499733 TI - [Recurrent tubal pregnancy after a intra-tubal injection of methotrexate. Analysis of an case report with a histological tubal study]. AB - A second tubal pregnancy occurred in the same tube as the first one which had been treated with methotrexate injection. The hysterosalpingogram performed 3 months after the methotrexate injection was normal. The recurrent tubal pregnancy was treated by laparoscopic salpingectomy. The pathology report confirmed non ruptured tubal pregnancy. No damage to the tube could be imputed to the methotrexate injection. This observation confirms that methotrexate has no deleterious effect on tubal epithelium but also emphasizes that recurrent ectopic pregnancy can occur despite a patent tube. PMID- 7499734 TI - [Treatment of endometrial clinical stage I adenocarcinoma by laparoscopic surgery. Seventeen cases]. AB - AIM: Study and follow-up of laparoscopic surgical treatment of endometrial cancer, clinical stage I. METHOD AND MATERIALS: This retrospective study covering the period 01 February 1990 to 31 January 1994 on 17 patients presenting with endometrial cancer, clinical stage I, that is, 35.4% of endometrial cancers operated in our department during that period, and for which laparoscopy was the first step in surgical treatment. RESULTS: Of the 17 cancers assessed by laparoscopy, 12 were also operated by laparoscopy. Conversion to laparotomy concerned 5 cases (29.4%): one case of postoperative adhesions, 2 cases of laparoscopic complications (cardiac rhythm problems, sub cutaneous emphysema), 2 cases of peritoneal granulations giving rise to suspicious of stage III. The mean age of the patients operated via laparoscopy was 59.9 years (extremes: 34-74). Their weight varied from 52 to 94 kg with a mean of 68.1 kg. Two endoscopic surgical complications arose (one bladder injury and one cervico-vaginal bleeding), but did not require conversion to laparotomy. No postoperative complications were noted. The average follow-up for these patients treated by endoscopy is 22.3 months. There was regrettably one early recurrence of the disease 7 months postoperatively. CONCLUSION: Laparoscopic surgery can be used to treat clinical stage I cases of endometrial cancer. This surgical approach must be used only for favourable cases and is reserved for teams trained in endoscopic and cancer surgery. PMID- 7499736 TI - [Intra-uterine artificial insemination. Indications, techniques, results; 83 cases]. AB - OBJECTIVES: Retrospective study of results after intra-uterine artificial insemination performed in the Poitiers University Hospital. MATERIAL: There were 95 infertile couples who benefited from intra-uterine artificial insemination. Indications involved pathology of the cervix in 53% of the cases and male infertility in 30%. Another cause of infertility was found in 5% and no cause could be identified in 12%. METHOD: The technique was as follows. Ovary stimulation using clomifene and hMG, sonographic monitoring with oestradiol assay, hCG for triggering ovulation, 2 inseminations at 24 and 48 h after swim-up sperm preparation and luteal phase supplementation. RESULTS: The overall success rate was 32.6% with a mean success rate per cycle of 11.1%. The best results were obtained in couples with unexplained infertility and cervix-related infertility (42 and 32% respectively). These results are comparable to those in the literature. CONCLUSION: The results of this series and the data reported in the literature demonstrate the interest of intra-uterine artificial insemination in cases of cervical sterility where it is indicated as first intention treatment. In case of impaired ejaculation, unexplained sterility and certain other causes of male infertility, intra-uterine artificial insemination can be useful but requires further study in order to determine the criteria for indications and the techniques to be used. Associating ovary stimulation increases the probability of success, especially in cervical sterility. Finally, more than 4 to 5 attempts do not increased success rate. PMID- 7499735 TI - [A Buschke and Lowenstein tumor in a female patient]. AB - Although it presents clinically as a malignant formation, the Buschke-Lowenstein tumour is known to be a histologically benign tumour caused by a papillomavirus infection from an condyloma acuminatum. The Buschke-Lowenstein tumour is generally observed in male subjects, usually on the penis, and rarely occurs in women. A female case is reported. In the literature, human papillomavirus 6 is the most common type although types 11, 16 and 18 are also reported. Surgery is the only treatment for this tumour which recurs readily. PMID- 7499737 TI - [Randomized study of sulprostone versus misoprostol in the cervical preparation before elective abortion in nulliparous women]. AB - OBJECTIVE: To assess the effectiveness and side effects of misoprostol versus suprostone in cervix preparation before aspiration abortion in the nullipara. METHODS: Ninety nulliparous women requesting medical abortion with gestations varying from 7 to 12 weeks were included in a prospective randomized study after informed consent. Women with contraindications for prostaglandins and those consulting before 7 weeks gestation were excluded from the study. The women were assigned to 30 groups randomly: women in group A were given 500 micrograms sulprostone in 250 saline solution infused at a rate of 100 micrograms/h 12 hours before the abortion; women in groupe B were given 400 micrograms misoprostol per os 3 hours before abortion; and women in groupe C were given 400 micrograms misoprostol per os 12 hours before abortion. RESULTS: The three groups were not different for age, parity or gestation, gestational age at abortion or type of anaesthesia. Dilatation of the cervix after treatment was identical in groups A and C (8.60 +/- 1.3 vs 8.20 +/- 1.3; NS) but was greater in group A than in group B (8.60 +/- 1.3 vs 7.23 +/- 2.2; p = 0.02). Maximal dilatation was the same in the three groups (9.50 +/- 0.8, 9.10 +/- 1.3, 9.17 +/- 0.8 respectively). Rate of partial expulsion was higher in groups A and C (23 and 6%) compared with group B (0%). Tolerance was much better with misprosol than with sulprostone: 43% digestive disorders in group A versus 15% in groupe B and C (p = 0.003); 80% pelvic pain with sulprostone versus 53% with misoprostol (p = 0.01); 70% metrorrhagie with sulprostone versus 38% with misoprostol (p = 0.04) and 36% with more bleeding than cycles in the sulprostone subjects versus 12% in the misoprostol subjects (p = 0.005). CONCLUSION: Misoprostol at a dose of 400 micrograms per os 12 hours before abortion in the nullipara is an interesting alternative to sulprostone. It is equally effective for cervix dilatation, tolerance is much better and cost is lower. PMID- 7499738 TI - [Isotretinoin (RoAccutane) embryopathy. A case report]. AB - Retinoids are synthetic vitamin A derivatives, particularly used in dermatology. Their prescription in women of childbearing age can cause, if pregnancy occurs, a serious malformative embryopathy, mainly involving external ear, brain and heart. A neonatal case caused by isotretinoin (RoAccutane) emphasizes the clinical and epidemiological data concerning this embryopathy. The aetiopathological hypothesis of an interaction between isotretinoin and Hox genes is advanced. Prophylactic measures are difficult since neonatal reported cases are uncommon, but antenatal exposition to this strong teratogenic agent results in multiple spontaneous abortions or pregnancy interruptions. PMID- 7499739 TI - [Doppler velocimetry in the umbilical, middle cerebral and aortic arteries, before and after cordocentesis]. AB - OBJECTIVE: Prospectively evaluate the effect of cordocentesis on the umbilical, fetal cerebral and aortic circulation. METHOD: Fetal blood was sampled for diagnostic purposes in 21 pregnant women at 21 to 38 weeks gestation. Ten patients undergoing amniocentesis served as controls. The resistance index (RI) in the umbilical and middle cerebral arteries and the mean blood velocity (Vm) in the descending aorta were measured with pulsed Doppler before and after blood sampling. Variations in umbilical and cerebral RI and in aortic Vm were recorded. RESULTS: There was a significant drop in both umbilical RI (mean +/- SD = -0.049 +/- 0.078; p = 0.009) and middle cerebral RI (-0.077 +/- 0.058; p < 0.0001) after cordocentesis. The drop in umbilical RI was greater when the second Doppler measurement was made early, when the blood was sampled transplacentally and in early gestational age. Reduction in fetal cerebral artery RI was also greater for transplacental puncture. The fetal descending aorta Vm did not change significantly after blood sampling. There were no variations in Doppler index before and after amniocentesis. CONCLUSIONS: Changes in blood flow velocity waveforms as measured by pulsed Doppler in the umbilical and fetal cerebral arteries can be induced by fetal blood sampling. Decreased resistance in the placenta and fetal circulation would imply release of nitric oxide. PMID- 7499740 TI - [Prognostic value of a single umbilical artery. 87 cases]. AB - OBJECTIVE: To study the effect of a single umbilical artery on infant prognosis. METHOD: Unicentric retrospective case-control study based on 18,425 births and semi-late pregnancy interruptions. RESULTS: There were 87 cases of single umbilical artery, frequently associated with a familial history of malformation in the sibhood (6.8%), gravid pathology (hypertension 11.5%, diabetes 6.8%), funicular anomaly (32%). One or several malformations were seen in 21% of the infants, the main cause of infant mortality (9%) and morbidity. The case-control study demonstrated that single umbilical artery corresponded to a moderate reduction in weight (-310 g), term (-1.4 weeks gestation), AGPAR scor at 1 and 5 minutes (-0.7) which was not significant when single umbilical artery was the sole pathology. CONCLUSION: When occurring alone, single umbilical artery has little effect on infant prognosis which is more dependant on associated malformations and gravid pathologies determining antenatal and postnatal management. PMID- 7499741 TI - [Placental chorioangioma. Two cases and review of the literature]. AB - Two cases of placental chorioangiomas were discovered, one echographically after in utero death at mid-term pregnancy and the second during the neonatal period associated with acute hydramniosis and a single umbilical artery. At 8 months, the new-born was in good health after persistent cholestasis at birth. A review of the literature of the macroscopic, histologic, echographic characteristics and the foetal-maternal complications of chorioangiomas revealed few cases of in utero death, especially so early in term. PMID- 7499743 TI - [Uncontrolled retrospective study of 75 pregnancies in women treated for epilepsy]. AB - OBJECTIVE: Our purpose was to study pregnancies and birth outcomes in a population of women treated for epilepsy. STUDY DESIGN: A retrospective uncontrolled study of 75 pregnancies in 55 epileptic women under treatment was conducted in the nursery of CHU Angers. The course of pregnancies and deliveries, the frequency of congenital malformations and the clinical state of the neonates during the first 6 days of life were recorded. RESULTS: There was a high rate of prematurity (15%) especially in patients treated by phenobarbital, a 10.5% rate of intra-utero growth retardation, a 11% rate of major congenital malformations and a 9% rate of isolated minor congenital malformations. Congenital malformations were mainly facial dysmorphia, cardiopahties, urogenital tract abnormalities. The neonatal symptomatology was associated with a high level of antiepileptic drugs in neonates or to a withdrawal syndrome or to hypovitaminosis. CONCLUSION: Overall results justify implementing prevention and detection measures. PMID- 7499742 TI - [Prevalence of hepatitis B viral markers in a population of pregnant women in Brazzaville (Congo)]. AB - The risk of perinatal transmission of hepatitis B virus is well-known; but, in Congo, where hepatitis B virus is endemic and the prevalence of hepatocellular carcinoma is high, there is no study on this mode of transmission. For three months, we screened systematically for HBsAg in 292 pregnant women who came as outpatients or to deliver in 3 health centres in Brazzaville. Positive sera for HBsAg were also tested for the other markers of HBV, except for specific DNA. The seroprevalence of HBV among these women was 6.5%. It was significantly higher among inpatients than among outpatients. The overall prevalence of HBs was 57.8%; the prevalence of the profile HBsAg + HBeAg was 2.05%. The risk of maternofoetal transmission of HBV was 2.7%. This study shows that the antenatal screening for HBV and the integration of the immunization against HBV in the expanded programme of immunization against HBV are good means to prevent the infection. PMID- 7499744 TI - [Pheochromocytoma and pregnancy. Two case reports]. AB - Pheochromocytoma is an uncommon tumour, young patients are involved. Diagnosis during pregnancy is scarcely made. Pheochromocytoma in pregnancy is a dramatic life threatening association for mother and foetus. High maternal and fetal morbidity and mortality can be reduced by a prompt diagnosis and treatment. We report two cases of pheochromocytoma associated with pregnancy, the prognosis of the first case was poor with stillbirth, the outcome was favourable in the second case for both mother and foetus. The authors emphasize the need of rapid diagnosis and treatment to improve outcome and the importance of perioperative management of these patients. PMID- 7499745 TI - [Unusual twin pregnancy: one in the pseudo-unicornate uterus and the other in the rudimentary uterine horn. Discussion on the diagnosis and management during pregnancy]. AB - An unusual twin pregnancy was diagnosed echographically at 18 weeks gestation and confirmed by magnetic resonance imaging. One foetus was in a pseudo unicornis uterus and the other in a rudimentary uterus cornu. The risk in such cases, as also reported in the literature, is rupture of the rudimentary cornu at about 20 weeks gestation. In this case the patient was carefully monitored to 23 weeks when the pregnancy in the rudimentary cornu stopped spontaneously. The foetus in the pseudounicornis developed normally to 38 week term. This exceptional observation emphasizes the risk of pregnancy in a blind uterus cornu. PMID- 7499746 TI - [Fetal cerebral accident due to massive fetomaternal hemorrhage. A case report]. AB - Massive fetomaternal haemorrhage (FMH) occurs in 0.12 to 0.5% of pregnancies. It is most often spontaneous and involves uncomplicated near-term pregnancies. It causes fetal anaemia, with or without fetal distress and hydrops fetalis. To our knowledge only one paper has reported a neurological complication (hemiplegia). We describe one case of FMH (maximal Kleihauer test = 6.5%) at 28 weeks gestation, which was spontaneous, reversible, associated with sinusoidal fetal heart rare (FHR) and hydrops fetalis; and complicated by an intraventricular antenatal haemorrhage at 30 weeks gestation. Echographic abnormalities decreased. The infant was born at 40 weeks gestation. Clinical examination was normal during the first week of life. At the age of 4 1/2 months, examination showed axial hypotonia and moderated dilatation of intracerebral lateral ventricules without any other brain damage. At the age of 24 months, the child had retarded walking and hypotonia. The outcome was spontaneously favourable with disappearance in utero of the intraventricular haemorrhage (HIV), without hydrocephalia or ischaemic lesions. Three cases of similar FMH have been reported but none of them described cerebral complications. Intrauterine intravascular transfusion should be proposed early. No single pathophysiological mechanism of FMH has been universally accepted and there is no aetiological treatment. The risk of recurrence of FMH in later pregnancies requires careful follow-up. PMID- 7499747 TI - [Spontaneous rupture of an unscarred uterus during labor and epidural anesthesia]. AB - We report a case of spontaneous rupture of unscarred uterus, during labour and under epidural analgesia in a 43 year-old patient at her third pregnancy and third delivery. The tear was vertical and took place on the left postero-lateral side of the uterus, occurring in the whole height of the lower segment and overlapping towards the uterine corpus and cervix. Analysis of medical, surgical, gynaecological and obstetrical past history has displayed no predisposing factors explaining this rare occurrence. This clinical case reminds us that spontaneous rupture is always possible even on an unscarred uterus. PMID- 7499748 TI - [Is there a relationship between a history of spontaneous abortion and low birthweight in Cotonou (West Africa)?]. AB - In Benin, as in other African countries, low birthweight infants is a major health problem with poorly understood causes. In a case-control study of 2114 mothers recruited before discharge from the "Lagune" maternity ward at Cotonou, we examined the relationship between spontaneous abortion and low birthweight infants in women who delivered between 15 June 1990 and 20 July 1991. Information was obtained by interview then on the basis of hospital files. The results indicated that there was no association between spontaneous abortion and premature birth: OR = 1.00 (95% CI = 0.61-1.56). Likewise, the relationship between a past history of spontaneous abortion and intrauterine growth retardation was not significant: OR = 1.19 (95% CI = 0.83-1.71). Adjustment with characteristics of the mothers did not improve the correlations. It is concluded that there is no relationship between a past history of spontaneous abortion and low birthweight. PMID- 7499749 TI - [Gastroschisis. Management. 50 cases]. AB - OBJECTIVE: Demonstrate the need for a multidisciplinary antenatal approach to laparoschisis. MATERIAL AND METHODS: A retrospective series of 50 children who underwent surgery for laparoschisis between 1975 and 1994 in the Infant Surgery Department of the Strasbourg University Hospitals. Postoperative outcomes were evaluated as a function of the stage of laparoschisis and surgical technique. Follow-up was also examined. RESULTS: Depending on the periods evaluated, ultrasound diagnosis varied from 16 to 92% and was made at 21 weeks gestation on the average. Lesions observed were atresia of the bowel (10%), complete intestinal involution (2%), and perivisceritis (88%). Complete parietal closure was achieved after bowel emptying in 58% of the newborns. Postoperative mortality, usually in vere severe forms, fell from 46% to 15% over the 20-years study period. CONCLUSION: Antenatal echography can confirm the diagnosis of laparoschisis. It can be used to define risk factors (size, bowel dilatation(s), mesenteric blood flow) allowing planned extraction in an Infantile Surgery Unit. This multidisciplinary attitude should help to improve overall outcome. PMID- 7499750 TI - [The indispensable necessity for informed consent before voluntary female sterilization]. PMID- 7499751 TI - [Amnio-infusion, a technique worth developing]. PMID- 7499752 TI - Neuroreality. I. Dedicated demolition of the decade of the brain:the genuine threat to neurologic research from the animal radical right American Academy of Neurology Animal Studies Subcommitte. PMID- 7499753 TI - Neuromuscular complications of sepsis in children. AB - Sepsis occurs frequently in the pediatric intensive care unit and is a significant cause of morbidity and mortality. Multiple organ systems are adversely affected by sepsis. Approximately 70% of adult patients with sepsis have peripheral nervous system dysfunction on electrophysiologic studies, of whom 30% are symptomatic. Neuromuscular dysfunction in children with sepsis is increasingly reported; however, the incidence remains undefined. Flaccid quadriplegia with the inability to wean from ventilatory support despite full cardiopulmonary recovery is the typical presentation. However, lesser degrees of weakness may be demonstrated with careful evaluation. Electrophysiologic studies often demonstrate the presence of axonal polyneuropathies, abnormalities of neuromuscular transmission, or acute myopathies. Identifiable neuromuscular syndromes in children with sepsis include critical illness polyneuropathy, pure motor polyneuropathy, thick-filament myopathy, and necrotizing myopathy. The common underlying pathogenic process in these syndromes appears to be sepsis, which may be accentuated by the administration of steroids or neuromuscular blocking agents. Recovery in strength usually occurs over a period of weeks to months. PMID- 7499754 TI - Clinical aspects of hereditary ataxias. AB - The hereditary ataxias are a group of complex genetic disorders the understanding of which is undergoing a revolution because of advances in molecular genetics. Within the last few years, at least seven different gene loci have been found to be responsible for these syndromes, and the search is on for additional loci that undoubtedly exist. This review summarizes the clinical features of the various hereditary ataxias with known gene loci, as well as others that are now defined on a clinical basis. It also deals with some of the imaging and neuropharmacologic advances that have been made in this group of disorders. PMID- 7499755 TI - Relapse of herpes simplex encephalitis. AB - We report five children who had recurrent central nervous system signs after conventional acyclovir therapy for herpes simplex encephalitis. Secondary exacerbation was characterized clinically by severe ballismic movement disorder in all five children, associated with fever, impairment of consciousness, and seizures. Biologic analysis in all children and magnetic resonance imaging and neuropathology studies of the brain in three cases were compatible with inflammatory reaction. In contrast, all viral cultures remained negative, herpes simplex virus antigen in one child and DNA tested by polymerase chain reaction in four children were undetectable in the first samples of cerebrospinal fluid during the relapse, suggesting a postinfectious, immune-mediated mechanism of relapse in these patients. PMID- 7499758 TI - Sudden unexpected death in patients with Dandy-Walker malformation. AB - An uncommon but well-recognized occurrence in patients with Dandy-Walker malformation is sudden unexpected death. The mechanism of demise has not been established. We report three patients with Dandy-Walker malformation that experienced sudden unexpected death without uncal or tonsillar herniation, the mechanism usually proposed for demise in such situations. Our findings suggest the possibility of vascular compromise as the cause of the sudden unexpected death in these patients. Early and effective relief of the pressure in the posterior fossa may prevent the occurrence of this catastrophic complication. PMID- 7499757 TI - Location of epileptic foci on interictal and immediate postictal single photon emission tomography in children with localization-related epilepsy. AB - Several neuroimaging techniques that supplement electrophysiologic methods of evaluating pediatric patients with localization-related epilepsies before surgery assess both structural and functional abnormalities. For example, single photon emission computed tomography (SPECT) with technetium-99m-hexamethylpropyleneamine oxime (99mTc-HMPAO) has been used to demonstrate abnormal cerebral perfusion. States of cerebral perfusion during the interictal and immediate postictal periods have been reported to correlate with epileptiform foci identified by electroencephalogram (EEG). Between January 1987 and March 1993, we studied 55 pediatric patients with intractable seizures with prolonged video EEG telemetry in the epilepsy monitoring unit, followed by computed tomography, magnetic resonance imaging, and SPECT, before surgery to determine whether SPECT studies with 99mTc-HMPAO improved the accuracy of locating the epileptic focus. Interictal SPECT was performed on all patients, and immediate postictal SPECT (within 10 minutes after seizure ended) on 17 patients monitored in the epilepsy monitoring unit. In 15 (88%) of the 17, the combination of interictal and postictal SPECT studies yielded results corresponding to the EEG abnormality, a result significantly better than that obtained from interictal studies alone: 21 (55%) of 38 (chi 2 = 5.647, P = .0175). SPECT scans showed localized abnormal perfusion in the ipsilateral temporal lobe in all six patients with mesial temporal sclerosis, but precise results were not obtained in cases of dual pathology and neuronal migration disorders. Depiction of cerebral perfusion by interictal and immediate postictal SPECT studies can lead to greater accuracy in the localization of epileptic foci. PMID- 7499756 TI - Ornithine transcarbamylase deficiency in females: an often overlooked cause of treatable encephalopathy. AB - Ornithine transcarbamylase deficiency is an X-linked recessive disorder of urea biosynthesis characterized by recurrent, often fatal, hyperammonemic encephalopathy in affected males; carrier females are usually asymptomatic. We report here the clinical and laboratory findings in five symptomatic heterozygous females with ornithine transcarbamylase deficiency. In each case, the onset of symptoms occurred in the 1st year of life, but diagnosis was delayed by up to 15 years. Symptoms included recurrent vomiting with lethargy (five patients), dietary protein intolerance (five), irritability (four), severe acute encephalopathy (three), ataxia (three), and acute hemiparesis (two). All eventually showed evidence of developmental delay or learning difficulties. Two of the three who experienced severe, acute, hyperammonemic encephalopathy suffered serious, permanent neurologic sequelae. Three of the patients showed decreased ornithine transcarbamylase activity in liver obtained by needle biopsy, and the other two had marked orotic aciduria associated with hyperammonemia. Neuroimaging studies demonstrated persistent abnormal lobar attenuation and abnormal signal on computed tomographic scan and magnetic resonance imaging. All patients showed marked symptomatic improvement on treatment with dietary protein restriction supplemented by pharmacologic measures to increase nonprotein nitrogen excretion. Ornithine transcarbamylase deficiency should be considered in the differential diagnosis of acute or chronic encephalopathy in females at any age. PMID- 7499759 TI - Models of delayed recovery. AB - After a biologic insult has impaired function of the developing central nervous system, recovery may not become apparent for years. Probability models adopted from the carcinogenesis, developmental neurobiology, learning decay, and stochastic process literatures are presented so that assumptions about apparent delays in the recovery process can be tested with data from longitudinal studies after a temporally circumscribed adverse event/exposure. This process of evaluating multiple models is exemplified with one data set. Nonlinear models of recovery are important because some children with early deficits first show improvement months to years later. PMID- 7499760 TI - The Oklahoma City bombing: a personal account. AB - On April 19, 1995, Oklahoma City (and the United States) lost its innocence. Almost all Oklahomans can relate exactly what they were doing either at 9:02 AM that day or when they first learned of the bombing. Of course, the whole world watched the events unfold through around-the-clock television coverage. One of the resident physicians in the University Hospital Emergency Medicine program, Dr Carl Spengler, was the first physician on the scene and directed early triage efforts. Because the Journal of Child Neurology is the only major biomedical journal with editorial offices in Oklahoma, we considered it appropriate that his personal account of this disaster be published in JCN. PMID- 7499761 TI - Peer review of the expert witness: an opportunity to improve our medical liability system. PMID- 7499762 TI - Rapid and fatal neurologic deterioration due to central nervous system Candida infection in an HIV-1-infected child. PMID- 7499763 TI - Brainstem tubers presenting as disordered breathing in tuberous sclerosis complex. PMID- 7499764 TI - Critical illness polyneuropathy in adolescence. PMID- 7499765 TI - Neonatal hemangiomatosis associated with brachial plexus palsy. PMID- 7499766 TI - Familial congenital hemiparesis. PMID- 7499767 TI - Teachers' awareness and opinion about AIDS: implications for school based AIDS education. AB - Two hundred thirteen teachers of one secondary and four primary schools of East Delhi were interviewed to assess their knowledge and attitudes about AIDS and their opinions regarding school based AIDS education. Observations revealed that majority of the teachers were aware of various aspects of HIV/AIDS. However, they also had some misconceptions regarding transmission of the disease. Though, most of the teachers opined that they could play an important role in educating the students as well as the community regarding AIDS/STDs, three fourths of them had never discussed AIDS/STDs with their students. Majority of teachers were in favour of starting class room based education on AIDS/STDs, beginning from secondary classes onwards, and more than half opined that class teacher could educate the students better than the doctors or parents. The results suggest that after being properly trained, teachers can be effectively utilized for educating the students. PMID- 7499768 TI - Profile of tuberculosis--an Army experience. AB - With the advent of HIV infection an increased awareness has arisen on its effect on the incidence of various diseases. In the Indian context an increased incidence of Tuberculosis has been expected. The Army is a controlled relatively healthy constituent of the general population. It hence provides an ideal sample for gauging any dramatic change in incidence. The present paper covers the secular trend of the disease from the days of independence and highlights the differing rates in the stratum of society within the Army. With increased awareness and improvement in diagnostic facilities an increased incidence of the disease has been noticed. However a prospective study to further study various features of the disease as well its correlation if any with HIV is necessary. The paper also touches upon manpower wastage and age profile of Tuberculosis in the Army, which is an important concern to them. PMID- 7499769 TI - Problems in the diagnosis of AIDS related cryptococcal meningitis: a case report. AB - Cryptococcal meningitis in a 35-year old male who had a history of chronic alcoholism and sexual promiscuity, is presented here. The patient presented twice, 6 weeks apart, with altered sensorium. On the earlier occasion, CSF examination and CT head scan were entirely normal, while on the second occasion CSF abnormalities were found which led to the diagnosis of cryptococcal meningitis. The patient eventually tested positive for HIV by both the ELISA and Western Blot techniques. Diagnostic problems and management difficulties in AIDS related cryptococcal meningitis are discussed. PMID- 7499771 TI - Need of an efficient surveillance to combat malaria. PMID- 7499770 TI - An outbreak of viral hepatitis A in Ibrahimpatnam town of Andhra Pradesh. PMID- 7499772 TI - Observation on host preference of Phlebotomus argentipes in district South-24 Parganas, West Bengal, India. PMID- 7499773 TI - DDT resistance in P. Papatasi in Bihar. PMID- 7499774 TI - A study of immune profile in human hydatid diseases. AB - A clinico haematological and immunological study was undertaken in 90 patients clinically suspected to be suffering from hydatid disease over a period of 1 year. The parameters studied included age of presentation, site of cyst localisation, haematological profile, total immunoglobins of different classes (IgG, IgM, IgA & IgE) and complement component C3, rosette forming lymphocytes, blast cell formation in response to phytohaemagglutinin (PHA-P) and concanavalin A (Con-A) and casoni test using standard methods. Twenty two out of 90 (22.44%) clinically suspected patients were surgically confirmed as hydatid disease cases. Hydatid disease occured in all age groups. Youngest case was 8 years and oldest 70 years. In 17/22 cases the cyst localised in the liver followed by lungs (3) neck (1) and kidney (1). Majority of patients (63.65%) belonged to blood group B. The mean total leucocyte and eosinophill counts were raised significantly (p < 0.001 and p < 0.05, respectively) in confirmed patients. The mean ESR value was raised in hydatid patients though, not significantly (P > 0.08). All the four classes of immunoglobulins viz. IgG, IgA, IgM, IgE and complement C3 were significantly raised in patients of hydatid disease compared to controls (P < 0.001, P < 0.002, p < 0.01 and p < 0.01, P < 0.02 respectively). The percentage of lymphocytes in peripheral blood in hydatid patients was reduced though, not significantly (P > 0.2). The absolute lymphocyte count was raised and mean percentage of T cells was reduced in patients with hydatid disease (P < 0.001 and P < 0.001, respectively).(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7499775 TI - Febrile episode among a floating population of C.R.P.F. Jawans stationed at Calcutta. AB - A virological investigation was carried out to establish the etiologic agent of a febrile outbreak amongst a floating population of C.R.P.F. Jawans, stationed at Calcutta during May-July, 1993. The illness was associated with fever, severe headache, bodyache and arthralgia which lasted for 2-4 days in most of the cases. Fifty cases were examined clinically and blood samples (both acute and convalescent) were subjected to virological study. One mouse-pathogenic agent could be isolated in suckling mice and identified as DEN-3. Sero-investigation of 50 single sera by HI & CF method revealed evidence of presumptive dengue infection in 33, while sero-conversion could be noted in 10 out of 25 available paired sera tested. Results of MAC ELISA revealed evidence of primary dengue infection in 6 out of 12 acute phase sera examined. Thus, the study shows that the episode is strongly suggestive of primary dengue infection. PMID- 7499776 TI - Transplacental passage of Plasmodium falciparum and seroevaluation of newborns in northern Nigeria. AB - The findings of a prospective study of 656 near-term pregnant women, and of the cord and peripheral blood of newborns of positive mothers are reported. 292 (44.51%) of the pregnant women were infected with P. falciparum. Further microscopic screening of the cord blood of newborns of the 292 positive cases at delivery showed a parasite rate of 10.95%. Transplacental passage of P. falciparum was confirmed by detection of parasitemia in the peripheral blood of 2.82% of newborns within 7 days of birth. Serological investigation of sera of 284 newborns by Indirect Fluorescent Technique (IFA) with P. falciparum IgM specific conjugate indicated that 72 (25.35%) had IgM antibodies of P. falciparum in their blood. The average birth weight of seropositive newborns was 400 gm less than seronegative ones. There was no significant difference in the rate of neonatal infection regardless of whether or not the mothers had taken chloroquine prophylaxis. PMID- 7499777 TI - Epidemiology of diarrhoea in two major cities in Saudi Arabia. AB - The epidemiological pattern of diarrhoeal diseases, causative agents and risk factors of their occurrence in two referral hospitals of Saudi Arabia is presented in this study. Stool specimens from 1726 admitted diarrhoeal cases were examined for parasites, yeast, enteropathogenic bacteria and rotavirus using the ELISA test. 41.3% of cases were due to rotavirus (RVGE) while 53.1% of cases showed no causative pathogens. Mean age of all cases was 20.2 months and RVGE cases showed a steady rise from the neonatal period onward, reaching a peak between 6-14 months. Males accounted for higher percentage of all diarrhoeal cases. Mothers of diarrhoea cases were mostly housewives with low educational level. Bottle fed children showed higher proportion (53.1%) of diarrhoea than other types of feeding suggesting the faeco-oral route of infection and the effect of poor sanitation. A pattern of higher RVGE cases was seen in warmer months in Al-taif and in cooler months in Jeddah. Findings demonstrated the interaction between host, pathogen and environmental factors in the epidemiology of infectious diarrhoeas in developing countries and the areas of possible prevention. PMID- 7499778 TI - An outbreak of viral hepatitis E: role of community practices. AB - A small localised outbreak of viral hepatitis due to HEV occurred in an educated and well placed community. The overall attack rate was found to be 1.9%; the children and adults were equally affected. No fatality was observed. Five blood samples collected from the cases of jaundice were found negative for Anti HAV IgM, HBsAg and Anti HBc IgM, but positive for Anti HEV. The infection spread by contamination of piped water by sewage system resulting from scarcity of water, intermittent water supply and installation of on-line private booster pumps by the residents. Community action, especially the boiling of drinking water till the quality of piped water improved, restricted jaundice cases to only one incubation period. The outbreak highlights the importance of community behaviour in first precipitating the crisis and then limitating the damage. PMID- 7499779 TI - Microbiological studies on cases diagnosed as typhoid/enteric fever in south-east Nigeria. AB - Eight hundred and nine patients suspected of having typhoid/enteric fever were investigated. Enteric fever bacilli (Salmonella typhi and S. paratyphi) were cultured from stool samples of 128 (16%) patients. Serological evidence of typhoid fever was obtained in 83 (13%) of 620 of the patients examined by the Widal test. The antibody titres ranged from 1:160 to 1:1280. A higher proportion of paratyphoid infection over typhoid infection was recorded both by culture (56%) and by the Widal test (63%). The predominant serotype was paratyphi C. Non typhoid Salmonellae were recovered from stool samples of 60 patients, comprising five serovars, namely, S. typhimurium-22 strains, S. enteritidis-15, S. hardar-9, S. virchow-5, S. bredeney-3, and 6 non-typable strains. Shigella sonnei and S. flexneri were detected in diarrhoeic and semi-formed stools of 19 (2.3%) of the typhoid suspects. It is clear from the results of the study that though typhoid is endemic in South-East Nigeria and the number of cases may rise at certain periods of the year, the news of a current wave of epidemic of typhoid seem unfounded. PMID- 7499780 TI - Temporary heterotopic auxiliary liver transplantation for fulminant hepatitis B. AB - BACKGROUND/AIMS: Orthotopic liver transplantation has been shown to improve survival in fulminant hepatic failure. However, after orthotopic liver transplantation life-long immunosuppression is necessary and graft complications may occur. METHODS: We employed heterotopic auxiliary liver transplantation in a 26-year-old man with fulminant hepatic failure due to hepatitis B virus infection. RESULTS: From a comatose state with seizures and decerebrate posturing, the patient woke up the day after heterotopic auxiliary liver transplantation. The graft functioned sufficiently. After 2 weeks, when peritonitis developed, immunosuppression was stopped since the native liver was recovering as shown by serial HIDA scans, liver biopsies, clotting parameters and serum bilirubin. When severe rejection of the graft developed 2 weeks later, and the peritonitis had been treated successfully, the native liver had recovered sufficiently to allow the graft to be removed. Now, more than 1 year after heterotopic auxiliary liver transplantation, the patient is free from medication and he is immune for hepatitis B virus, his liver tests have returned to normal and he has regained his normal life. CONCLUSIONS: Temporary heterotopic auxiliary liver transplantation for fulminant hepatitis B is feasible. PMID- 7499781 TI - Failure of ursodeoxycholic acid to prevent acute cellular rejection after liver transplantation. AB - BACKGROUND/AIMS: Acute rejection is still a major problem after liver transplantation. Ursodeoxycholic acid has beneficial effects in cholestasis by reducing the expression of major histocompatibility complex antigens. METHODS: We have performed a double-blind randomised study comparing ursodeoxycholic acid with placebo for the prevention of acute cellular rejection after liver transplantation. Twenty-six patients received ursodeoxycholic acid 600 mg per day and 24 patients received placebo for 2 months. RESULTS: Neither rejection incidence nor rejection severity was significantly different in the two groups (p > 0.90). CONCLUSIONS: We conclude that adjuvant ursodeoxycholic acid administration does not prevent rejection after liver transplantation. PMID- 7499782 TI - Analysis of prognostic variables in the prediction of mortality, shunt failure, variceal rebleeding and encephalopathy following the transjugular intrahepatic portosystemic stent-shunt for variceal haemorrhage. AB - BACKGROUND/AIMS: The aim of this study was to analyse prognostic variables predicting mortality, shunt insufficiency, variceal rebleeding and encephalopathy following transjugular intrahepatic portosystemic stent-shunt for variceal haemorrhage. METHODS: Sixty-eight patients with cirrhosis who successfully underwent transjugular intrahepatic portosystemic stent-shunt for variceal haemorrhage were studied. Mean age was 54.8 years (s.e. 1.5); 41 were males and 27 were females. Mean Child score was 8.3 (s.e. 0.3). Cirrhosis was alcohol related in 47. The patients have been followed up for a mean of 10.8 (s.e. 1.1) months. The univariate association between 30 prognostic variables and post treatment encephalopathy was tested by the Chi-squared or Wilcoxon rank-sum tests and multiple logistic regression was used to test the significance of factors adjusted for one another. Cox's proportional hazard regression was used to test the univariate and multivariate significance of the other three outcomes. RESULTS: Transjugular intrahepatic portosystemic stent-shunt was performed successfully in 89.5% of patients. Twenty-two patients have died and eight have been transplanted. Fourteen patients died within 30 days of transjugular intrahepatic portosystemic stent-shunt and death was predicted independently by hyponatraemia (p < 0.001) and by severe liver disease (Child C, p < 0.001). Eight patients died during follow up and survival in the long term was predicted independently by the presence of encephalopathy (p < 0.001) prior to transjugular intrahepatic portosystemic stent-shunt (p < 0.001). Shunt insufficiency was predicted by an initial portal pressure gradient of greater than 18 mmHg (p < 0.01). None of the variables analysed predicted variceal rebleeding. Encephalopathy following transjugular intrahepatic portosystemic stent-shunt was only predicted indepedently by its presence prior to transjugular intrahepatic portosystemic stent-shunt (p < 0.001). CONCLUSIONS: The results of this study suggest that patients with severe liver disease and hyponatraemia are liable to die early, and the presence of encephalopathy prior to transjugular intrahepatic portosystemic stent-shunt independently determines long-term survival. Patients in these groups should be considered high risk and worked up for orthotopic liver transplantation early. Shunt function in patients with an initial portal pressure gradient of > 18 mmHg requires close supervision. Encephalopathic patients should have smaller shunts and prophylactic measures to prevent worsening encephalopathy. PMID- 7499783 TI - Functional hepatic flow and Doppler-assessed total hepatic flow in control subjects and in patients with cirrhosis. AB - Functional hepatic flow and total hepatic flow were determined by non-invasive techniques in 32 patients with cirrhosis and in 32 paired control subjects. Functional hepatic flow was measured by the hepatic clearance of D-sorbitol, while total hepatic flow was determined by pulsed echo-Doppler, as the sum of portal and hepatic arterial blood flow. Functional hepatic flow was significantly reduced in patients with cirrhosis (927 +/- 314 vs. 1287 +/- 315; p < 0.0001), while total hepatic flow was slightly increased (1511 +/- 540 vs. 1261 +/- 321 in controls; p = 0.028). In control subjects functional hepatic flow significantly correlated with total hepatic flow (r = 0.823; p < 0.001), while no correlation was observed in cirrhosis. Functional hepatic flow and the difference between total hepatic flow and functional hepatic flow significantly correlated with the Child-Pugh score in patients with cirrhosis. The data obtained in control subjects support the measurement of functional hepatic flow and total hepatic flow by non-invasive techniques. The finding that in cirrhosis functional hepatic flow is significantly decreased, while Doppler-assessed total hepatic flow is preserved or even increased, confirms that a relevant part of blood flowing through the liver is diverted by intrahepatic shunts. The simultaneous assessment of these two parameters by non-invasive techniques may be proposed as a reliable tool for the study of functional shunting of cirrhosis. PMID- 7499784 TI - Endothelin-1 and endothelin-3 in cirrhosis: relations to systemic and splanchnic haemodynamics. AB - BACKGROUND/AIMS: Endothelins are isopeptides with potent vasoactive properties, but their implications in the hyperkinetic syndrome in cirrhosis are obscure. Therefore, the aim of the present study was to relate hepatic venous and circulating endothelin-1 and endothelin-3 to systemic and splanchnic haemodynamics. METHODS: Endothelin-1 and endothelin-3 were measured in samples from a hepatic vein and the femoral artery in 42 patients with cirrhosis, eight hypertensive controls and 10 normotensive controls. RESULTS: Hepatic venous endothelin-1 was significantly higher in the patients with cirrhosis, mean 21.2 +/- 0.9 pg/ml (SEM) than in the hypertensive controls, 12.4 +/- 2.4 pg/ml, and normotensive controls, 9.6 +/- 1.6 pg/ml (p < 0.00001). Similarly arterial endothelin-1 was significantly higher in the patients with cirrhosis than in the controls (p < 0.00001). Hepatic venous endothelin-1 was significantly correlated with the hepatic venous pressure gradient (r = 0.61, p < 0.00004), serum creatinine (r = 0.35, p < 0.03), diastolic blood pressure (r = -0.31, p < 0.05), central and arterial blood volume (-0.36, p < 0.05), central circulation time (r = -0.41, p < 0.02), and serum sodium (r = -0.56, p < 0.00002) in the patients with cirrhosis. The hepatosplanchnic release of endothelin-1, assessed as the arteriohepatic-venous difference adjusted for hepatic plasma flow, was higher in the group with cirrhosis, 1.5 +/- 0.4 ng/min, than in the normotensive controls, 0.1 +/- 0.2 ng/min (p < 0.01), and was furthermore correlated to the cardiac output in the group with cirrhosis (r = 0.35, p < 0.04). Hepatic venous endothelin-3 was higher in the patients with cirrhosis, 19.0 +/- 1.4 pg/ml (n = 23), as compared with hypertensive controls, 14.2 +/- 1.3 pg/ml, and normotensive controls, 10.0 +/- 1.4 pg/ml (p < 0.002). The same pattern was found in arterial endothelin-3. Hepatic venous endothelin-3 correlated significantly with central and arterial blood volume (r = 0.56, p < 0.02). The hepatosplanchnic release of endothelin-3 was higher in the patients with cirrhosis, 1.0 +/- 0.7 ng/min, than in the normotensive controls, -0.7 +/- 0.4 ng/min (p = 0.05). CONCLUSIONS: In the presence of cirrhosis, hepatic venous and circulating endothelin-1 and endothelin 3 are elevated with significant relations to systemic and splanchnic haemodynamics, and the hepatosplanchnic release of both peptides is increased. This suggests that the endothelin system is implicated in both systemic and portal haemodynamic abnormalities in cirrhosis, although this study does not allow conclusions on causal relationships. PMID- 7499785 TI - Serum markers for connective tissue turnover in patients with chronic hepatitis B and chronic hepatitis C: a comparative analysis. AB - BACKGROUND/AIMS: Clinical and pathological differences between chronic hepatitis B and chronic hepatitis C have now been established. METHODS: To compare hepatic connective tissue metabolism in chronic hepatitis B and C, we determined serum levels of prolyl 4-hydroxylase beta-subunit, procollagen III aminoterminal peptide, type IV collagen 7S domain, the central helix region of type IV collagen, tissue inhibitor of metalloproteinase, and hyaluronan in 55 patients with chronic hepatitis B and 83 patients with chronic hepatitis C. RESULTS: There were no significant differences in the above markers for connective tissue turnover between the chronic hepatitis B group and the chronic hepatitis C group. CONCLUSIONS: These results suggest that the accelerated connective tissue metabolism observed in chronic viral hepatitis is independent of the causative virus. PMID- 7499786 TI - Effects of ethanol on hepatitis B virus Pre-S/S gene expression in the human hepatocellular carcinoma derived HEP G2 hepatitis B DNA positive cell line. AB - BACKGROUND/AIMS: Among the reported interactions between ethanol and hepatitis B virus (HBV), studies of transgenic mice have suggested an effect of ethanol on the secretion of viral envelope proteins. METHODS: We further investigated these interactions in vitro by determining HBs antigen levels and performing northern blots of viral mRNA in human cell culture (HepG2 HBV positive cells) exposed for 3 to 12 days to various concentrations of ethanol. RESULTS: In cultures exposed to 200 mM ethanol, HBs antigen concentrations increased in the medium (p < 0.05) after 3 days as Pre-S1 and Pre-S2 protein concentrations. This increase was not specific, as albumin and ferritin increased in the same proportions. Ethanol also increased the HBs antigen concentration in the cells (p < 0.05), whereas levels of viral mRNA encoding surface proteins were unaffected. CONCLUSIONS: These findings show that short-term ethanol exposure in vitro can induce HBs antigen overexpression via a post-transcriptional mechanism. PMID- 7499787 TI - Parathyroid hormone-related peptide is expressed and rapidly inducible in human liver cell cultures that have a bile duct phenotype. AB - Parathyroid hormone-related peptide is the major factor responsible for hypercalcemia of malignancy. There is increasing evidence that parathyroid hormone-related peptide also plays an important role in the growth and differentiation of both neoplastic and non-neoplastic cells. Recently we found that reactive human bile ductules and cholangiocarcinomas, but not normal bile ducts, human hepatocytes nor hepatocellular carcinomas, express parathyroid hormone-related peptide and we speculated that parathyroid hormone-related peptide may function as a growth and differentiation factor for bile ductular epithelial cells. Using a specific polyclonal antibody for immunostaining and a digoxigenin-random prime-labeled probe for in situ hybridization assay, we found that only cell lines with a bile duct phenotype expressed parathyroid hormone related peptide and its mRNA. HepG2 cells with hepatocellular phenotype (CK19-, CK7-, CK8+, CK18+, albumin+) do not express parathyroid hormone-related peptide. However, A16 (HepG2 derived cell line) expressing bile duct marker CK19, also expressed parathyroid hormone-related peptide, while hepatocyte markers CK8, CK18, CALLA and albumin were negative. In addition, the H1 cell line (adult human hepatocytes immortalized in our laboratory by SV40 DNA transfection, passaged at least 40 times and cultured for 13 months) expressed bile duct marker CK7 and parathyroid hormone-related peptide, while hepatocyte markers CK8, CK18, CALLA and albumin were negative. Previous studies demonstrated that parathyroid hormone related peptide gene expression in keratinocytes can be modulated by serum, growth factors and cycloheximide although there is a species and cellular specificity.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7499788 TI - Inhibition of iron toxicity in rat and human hepatocyte cultures by the hydroxypyridin-4-ones CP20 and CP94. AB - The protective effect of the hydroxypyridin-4-ones (CP20 and CP94) was studied on iron-loaded rat and human hepatocytes; desferrioxamine B was used as a chelator reference. Iron load was achieved by addition of 5 up to 50 microM iron citrate to the culture medium. One day after iron treatment, an increase in lactate dehydrogenase, aspartate aminotransferase, alanine aminotransferase and malondialdehyde extracellular concentrations was measured in rat and human hepatocyte cultures. This enzyme release and the increase in free extracellular malondialdehyde were observed with 5 microM iron and high levels were obtained with 50 microM. The bidentate chelators CP20 and CP94 (150 microM) appeared to be as effective as the hexadentate chelator desferrioxamine (50 microM) in the protection of rat and human hepatocytes against the toxic effect of iron load achieved by culturing the cells for 1 day in the presence of 50 microM iron citrate. In rat and human hepatocytes cultured for 1 day in the presence of 1 microM 55Fe-50 microM iron citrate plus CP20, CP94 or desferrioxamine B, a decrease of iron uptake by the cells was observed. When the hepatocytes were cultured for 1 day in the presence of 1 microM 55Fe-50 microM iron citrate and then for a further day in the presence of CP20, CP94 or desferrioxamine B but not iron, the chelators decreased the intracellular iron level, indicating their iron releasing effect from the loaded cells. The observed effects of the hydroxypyridin-4-ones CP20 and CP94 were as potent as the effect of desferrioxamine B. This study presents new data favoring the potential clinical interest of this new class of chelating agents in the treatment of human iron overload. PMID- 7499789 TI - Expression and phosphorylation of rat c-met/hepatocyte growth factor receptor during rat liver regeneration. AB - Hepatocyte growth factor receptor is identified as a heterodimeric tyrosine kinase encoded by the c-met gene. This study was designed to determine how the c met/hepatocyte growth factor receptor participates in the intracellular events involved in rat liver regeneration induced by administration of carbon tetrachloride. Expression of the rat c-met mRNA increased, peaking 24 h after carbon tetrachloride administration almost in parallel with MET protein expression. Histochemical studies demonstrated that expression of the rat c-met was enhanced in cells surrounding the damaged areas, and also that the distribution of cells expressing MET was almost in accordance with that of cells expressing proliferating cells nuclear antigen. The MET protein underwent intense tyrosine phosphorylation peaking at 12 h after carbon tetrachloride administration, and prior to DNA synthesis. Phospholipase C gamma and phosphatidylinositol 3-kinase, intracellular signal transducing molecules containing Src homology 2 domain, were associated with the MET protein following tyrosine phosphorylation in vivo. These observations suggest that expression and tyrosine phosphorylation of MET protein associated with signal transducing molecules may provide a mechanism whereby hepatocyte growth factor exerts its action on hepatocyte growth during rat liver regeneration induced by carbon tetrachloride administration. PMID- 7499792 TI - Fulminant hepatitis caused by a hepatitis B virus core region variant strain. AB - We studied the viral genome of a hepatitis B viral strain isolated from a patient with fulminant hepatitis. The patient was followed from prior to the rise in transaminases until she recovered. The precore and core regions of the viral strains were sequenced before and after the illness via the polymerase chain reaction and subcloning methods. Prior to her clinical illness, a strain with precore wild-type sequence and core mutations corresponding to amino acid residues 77 and 113 was noted in large quantities. With the onset of hepatitis, this core variant completely disappeared. Very low titers of precore and core wild or partial core deletion strains remained 1 month later. The core variants described may have contributed to the severe host immune reaction, fulminant hepatitis and immune-mediated viral clearance. Such variants appeared to have been eliminated, and wild and core-deleted virus that lacked the peculiar mutations remained. PMID- 7499791 TI - Time-course changes in gastric mucosal blood perfusion following portal vein constriction in rats. AB - BACKGROUND/AIMS: Gastric mucosal blood flow is increased in all experimental models of chronic portal hypertension, when portosystemic shunting and the hyperdynamic circulation are fully developed. However, some controversy exists concerning the time course of this event. This study was undertaken to investigate the chronological changes in gastric mucosal blood perfusion during the first 7 days after partial portal vein constriction. METHODS: Portal hypertensive and sham operated animals were studied. Gastric mucosal blood flow was measured by hydrogen-gas clearance and gastric oxygen and hemoglobin content by reflectance spectrophotometry, prior to and immediately after partial portal vein constriction, and 1, 2, 7 and 15 days after induction of portal hypertension. RESULTS: Immediately after partial portal vein constriction, gastric mucosal blood flow and gastric oxygen were significantly decreased by 27% and 32% respectively, and gastric hemoglobin significantly increased by 19%. On day 1, gastric mucosal blood flow was increased in both portal hypertensive and sham operated rats. However, while this parameter returned to normal in sham operated rats, it remained high on days 2, 7 and 15 in portal hypertensive rats. In portal hypertensive rats, gastric oxygen values were not significantly different from those in sham operated animals, but hemoglobin tended to increase with development of portal hypertension, being significantly higher than in sham operated rats at days 2 and 7. Similar results were obtained when analyzing gastric mucosal blood perfusion 2 and 7 days after two-staged total portal vein constriction. CONCLUSIONS: Increased gastric mucosal blood perfusion is present in portal hypertensive rats from the first day after partial portal vein constriction. The lack of significant differences in all parameters on day 1 between portal hypertensive and sham operated rats is probably related to an increased mucosal blood perfusion related to surgery. PMID- 7499790 TI - Functional activity of isolated pig hepatocytes attached to different extracellular matrix substrates. Implication for application of pig hepatocytes in a bioartificial liver. AB - For the manufacture of a bioartificial liver for human application, large amounts of viable and active hepatocytes are needed. Pig hepatocytes are considered to be the best alternative to scarce human hepatocytes. In vitro hepatocyte functions have so far been tested under different circumstances, mainly with rat hepatocytes. Pig hepatocytes were isolated with a single two-step isolation procedure, resulting in a high yield of viable hepatocytes. The hepatocytes were tested for their ability to synthesise urea, to metabolise 7-ethoxycoumarin (cytochrome P450 activity), and to synthesise and secrete proteins. These activities of hepatocytes while attached to tissue culture plastic were compared to the activity of the cells attached to several extracellular matrix constituents: collagen I and IV, laminin, fibronectin, Engelbreth-Holm-Swarm Natrix and in the presence of Matrigel. With the exception of Matrigel, neither of the extracellular matrix substrates enhanced pig hepatocyte function compared to tissue culture plastic. However, relatively large amounts of murine proteins leak out of the Matrigel. The advisability of using Matrigel or other extracellular matrix proteins in a bioartificial liver loaded with pig hepatocytes is discussed. PMID- 7499793 TI - Autoimmune hepatitis following hepatitis A virus infection. AB - A 7-year-old patient is reported who suffered from fatigue and jaundice due to chronic hepatitis. He had acquired hepatitis A virus infection in his community and communicated the disease to his German family 4 weeks later. While the other family members recovered from acute viral hepatitis A, the patient presented 10 weeks after the onset of hyperbilirubinemia (12 mg/dl) with the histology of chronic hepatitis, absence of markers for viral persistence, presence of autoantibodies against smooth muscle (1:320) and the asialoglycoprotein receptor (1:600), and marked hypergammaglobulinemia (3700 mg/dl), leading to the diagnosis of autoimmune hepatitis. The patient received immunosuppressive therapy, symptoms of liver disease disappeared, and autoantibodies cleared from circulation. The case is discussed in the context of a putative virus-induced autoimmune hepatitis in childhood. Autoimmune hepatitis may be induced by an external trigger. Hepatitis A virus infection is one of probably several triggers that may induce autoimmune hepatitis in predisposed individuals. PMID- 7499794 TI - Reversible AIDS-related sclerosing cholangitis. AB - Although hepatobiliary involvement is common in the acquired immunodeficiency syndrome, it infrequently leads to biliary tract abnormalities. We describe a 39 year-old man with human immunodeficiency virus infection and no previous acquired immunodeficiency syndrome-defining illnesses, who presented with malaise, right upper quadrant pain, lymphadenopathy and cholestasis. An endoscopic retrograde cholangiopancreatography demonstrated sclerosing cholangitis due to disseminated B-cell nonHodgkin's lymphoma. Following chemotherapy, his symptoms and signs rapidly improved, so that 1 month later his endoscopic retrograde cholangiopancreatography had returned entirely to normal. PMID- 7499795 TI - Functional loss of cerebral blood flow autoregulation in patients with fulminant hepatic failure. AB - In management of patients with fulminant hepatic failure, it is recommended that mean arterial pressure should be raised if cerebral perfusion pressure is lower than 50 mmHg, but the influence of such therapy on cerebral blood flow is unknown. We examined cerebral blood flow autoregulation in seven consecutive patients with fulminant hepatic failure during treatment of imminent insufficient cerebral perfusion pressure. Cerebral perfusion was evaluated by transcranial Doppler assessed mean flow velocity in the middle cerebral artery and by the arterio-venous difference for oxygen. Intracranial pressure was recorded by a subdural transducer and cerebral perfusion pressure calculated as the difference between mean arterial pressure and intracranial pressure. After 20 (range 10 to 43) min, mean arterial pressure was raised from 74 (43-80) to 94 (76-114) mmHg by i.v. noradrenaline, cerebral perfusion pressure increased from 49 (26-75) to 82 (50-108) mmHg (p < 0.01) as the intracranial pressure remained unchanged at 26 (3 35) mmHg. The mean flow veolocity increased from 68 (30-134) to 108 (48-168) cm s 1 and the arterio-venous difference for oxygen by 46 (10-82)% (p < 0.05). Both mean flow velocity (r = 0.63) and arterio-venous difference for oxygen (r = 0.71) were correlated to mean arterial pressure (p < 0.001), and a lower blood pressure limit of autoregulation could not be identified in any of the patients. These data suggest that the cerebral blood flow is not autoregulated in patients with fulminant hepatic failure and therefore cerebral blood flow should be "clamped" within the normal physiologic range by manipulation of arterial blood pressure in order to avoid cerebral hypoxia and/or hypertensive induced cerebral oedema. PMID- 7499796 TI - The role of nitric oxide in the hyperdynamic circulatory syndrome associated with portal hypertension. PMID- 7499797 TI - Adhesion molecules and liver transplantation: new strategies for therapeutic intervention. PMID- 7499799 TI - Thrombocytopenia of chronic liver disease corrected by erythropoietin treatment. PMID- 7499800 TI - Low prevalence of antibodies to hepatitis C virus in patients with Hashimoto's thyroiditis. PMID- 7499798 TI - Erythropoietin treatment of thrombocytopenia of chronic liver disease: need for more strictly-defined inclusion and evaluation criteria. PMID- 7499801 TI - Comparison of clinical judgment and anthropometric parameters for evaluating nutritional status in patients with alcoholic liver disease. PMID- 7499803 TI - Pool tests for detection of hepatitis C virus infection. PMID- 7499802 TI - Vitamin K1 improves survival in bile-duct-ligated rats with cirrhosis. PMID- 7499804 TI - Survey of operating theatres in Great Britain and Ireland. AB - A survey of operating theatres in Great Britain and Ireland by postal questionnaire was undertaken to determine the number of non-ventilated theatres in use, the number of designated theatres for specialist surgery and how and when bacterial sampling is conducted. Replies were received from 147 centres covering 438 operating theatre suites. Eighty-seven (59%) centres contained three or fewer suites and only 32% did not have a designated theatre for any specialist surgery. One hundred and seventy-three (40%) theatre suites were built over 20 years ago and 28 non-ventilated theatres were identified, mainly used for minor surgery (e.g. removal of 'lumps'). Four percent of plenum and 9% of ultraclean theatres are never monitored bacteriologically and settle plates are used in 72 (49%) centres. This survey suggests there is some confusion over the indications for bacteriological monitoring and what constitutes acceptable standards. The use of non-ventilated theatres, except for the most minor of procedures, is of some concern and should be phased out. PMID- 7499805 TI - Infections following epidural catheterization. AB - Seventy-eight patients with culture-positive epidural catheters, were studied. Fifty-nine had symptoms of exit site infection and 11 patients had clinical meningitis, two of whom also had an epidural abscess. This corresponds to a local infection incidence of at least 4.3% and an incidence of central nervous system infection of at least 0.7% at Odense University Hospital. This degree of infection is of the same magnitude as that reported for intravascular devices. We found that the patients with generalized symptoms of infection had been catheterized for a longer time, and were older than patients with only local symptoms of infection. The microorganisms isolated from the tips of the epidural catheters were coagulase-negative staphylococci (41%), Staphylococcus aureus (35%), Gram-negative bacilli (14%) and others (10%). The Gram-negative bacilli and S. aureus caused serious infections more frequently than the others. We discuss the symptoms and diagnosis of spinal epidural abscess and suggest a proposal for prophylactic and diagnostic guidelines for epidural catheter-related infections. PMID- 7499806 TI - Subtyping of Shigella sonnei for tracing nosocomial transmission. AB - Nosocomial transfer of Shigella spp. occurred between two patients sharing the patient room and toilet in a ward for internal medicine. Both patients had travelled abroad shortly before the onset of illness. Based on the epidemiology of Shigella sonnei strains imported into Finland, nosocomial infection was strongly suspected. The isolates of S. sonnei were subtyped using ribotyping, plasmid analysis and antibiotic resistance patterns. The greatest discrimination between the strains was achieved by ribotyping. The results of the typing scheme confirmed the nosocomial transmission of S. sonnei. The outbreak did not spread further within the ward. The importance of isolation of a patient with diarrhoea in a hospital setting is emphasized. PMID- 7499807 TI - Identification and biotyping of Acinetobacter spp. isolated in Chilean hospitals. AB - Two hundred and eighty-one isolates of Acinetobacter spp. obtained from clinical specimens in hospitals from five Chilean cities were identified to species level and biotyped. Respiratory tract and wound secretions were the main sources of the isolates. Acinetobacter baumannii was the most frequent species (96.8%), followed by Acinetobacter genospecies 3 (2.8%). Twelve different biotypes of A. baumannii strains were found of which biotypes 9, 8 and 6 were the most frequent. Isolates of other biotypes were rare. These results differ from most other Latin American and European countries. PMID- 7499808 TI - Hepatitis B virus, hepatitis C virus and human immunodeficiency virus infection in health care workers: a multiple regression analysis of risk factors. AB - A seroprevalence survey of hepatitis B virus (HBV), hepatitis C virus (HCV) and human immunodeficiency virus (HIV), was conducted using serum samples obtained from 5813 health care workers (HCWs) in five public hospitals in the Latium region of Italy, during the 1985 vaccination campaign against HBV. The seroprevalences of HBV, HCV and HIV were 23.3% [95% confidence interval (CI) = 22.3-24.4%], 2% (95% CI = 1.6-2.4%) and 0.07% (95% CI = 0.001-0.13%), respectively. In a logistic regression model, sex, increasing age, all job categories vs. physicians, dental treatment in the previous six months, and needlestick injury during the previous year were significantly associated with HBV. Conversely, no occupational and community risk factors, but only history of blood transfusion were significantly associated with HCV. Nevertheless, the documented risk of HCV as well as of HIV transmission through percutaneous and mucocutaneous exposure to blood and body fluids should lead to continued efforts to minimize risks of infection by enhancing the compliance of HCWs with vaccination against HBV and adherence to infection control measures, and by introducing safer devices and techniques. PMID- 7499809 TI - The effects of 'in-use' surgical handwashing on the pre- and postoperative fingertip flora during cardiothoracic and orthopaedic surgery. AB - Two operating teams (25 persons) were followed for two months with fingerprint samples taken preoperatively; before and after 'in-use' surgical handwashing; and immediately postoperatively, with and without surgical gloves. The mean time for handwashing for the cardiothoracic team (CT) was 2 min and for the orthopaedic team (OT) was 3.5 min. A closer observation of 10 persons revealed a great individual variation in washing techniques, in spite of standard guidelines. The CT team performed eight, and the OT team nine sterile operations with an average duration of 3 h and 20 min and 2 h and 40 min, respectively. Surgical handwashing resulted in fingertip sterility in 111/118 (94.1%) cases; in 61/66 (92.4%) samples from the surgeons and in 50/52 (96.2%) samples from the assistants. Postoperative fingerprinting with gloves on showed sterile conditions in 85/91 (93.4%) samples; 57/59 (96.6%) from the surgeons and 28/32 (87.5%) from the assistants. Immediately after removal of the gloves, 43/67 (64.2%) of fingerprint samples from the surgeons and 13/48 (27.1%) from the assistants were still sterile. Coagulase-negative staphylococci (CNS) and Bacillus species predominated in fingerprint samples. Of the 105 CNS strains tested, 11.4% were methicillin resistant. Only five strains of Staphylococcus aureus were isolated; in 4/5 cases from the OT. This study illustrates that in spite of standard guidelines, there is great individual variation in surgical handwashing. However, in most instances, the bacteria are eradicated from the fingertips. Even after surgery for 2-3 h, there may still be a residual effect of the hand disinfecting agent in half of the cases. PMID- 7499810 TI - Evaluation of three glutaraldehyde-based disinfectants used in endoscopy. AB - The European suspension test was applied to compare the in vitro activity of three glutaraldehyde-based disinfectants: a 1:10 dilution of a 10% glutaraldehyde solution containing 0.5% phenylphenol-0.1% amylphenol, a 2% acid glutaraldehyde solution, and a 2% alkaline glutaraldehyde solution. The microbicidal effect of the disinfectants was evaluated by counting surviving cells of three indicator microorganisms (Pseudomonas aeruginosa, Mycobacterium chelonae and phage f2) after exposure times of 5, 10, 20 and 40 min to the agents at 20 degrees C. An inactivation factor (IF) of > or = 5 log10 was used as the criterion for effective disinfection. This IF was achieved with every microorganism/disinfectant combination after 5 min exposure except in experiments with phage f2 and the 1% glutaraldehyde-based disinfectant. The diminished inactivation noticed with the 1% glutaraldehyde-based disinfectant supports the recommendation to use a disinfectant containing a minimum of 2% glutaraldehyde for high level disinfection. PMID- 7499811 TI - A survey of glove preferences of general and orthopaedic surgeons in North West Thames Regional Health Authority. AB - The glove preferences of all the consultant surgeons and consultant orthopaedic surgeons in North West Thames were surveyed by questionnaire. Responses were received from 59 of 78 general surgeons and 47 of 55 orthopaedic surgeons. Twenty two percent of general surgeons never double gloved compared with 4% of orthopaedic surgeons (P < 0.01). Seventy-three percent of general surgeons double gloved for 'high risk' patients compared with 17% of orthopaedic surgeons (P < 0.001). When double gloving the majority of surgeons (67%) used their normal sized glove inside and half a size larger on the outside. PMID- 7499812 TI - Nosocomial and community-acquired Xanthomonas maltophilia infection in tropical Australia. AB - Xanthomonas maltophilia infection is recognized as a serious problem in association with immunosuppressive and invasive therapies, and with the use of broad-spectrum antibiotics. In Darwin Hospital in Australia's Northern Territory preliminary evidence of nosocomial transmission of X. maltophilia prompted this retrospective examination of all X. maltophilia isolates over a 30 month period. X. maltophilia was most frequently isolated in the 'wet season' corresponding to times of increased antibiotic treatment of the serious community-acquired pneumonias encountered in this tropical region. A relatively high proportion of community-acquired isolates (4/18; 22%) was documented. This study demonstrates that X. maltophilia infection is an emerging cause of morbidity in tropical regions where endemic infections require the use of broad-spectrum beta-lactams. PMID- 7499813 TI - The neonatal laryngoscope as a potential source of cross-infection. PMID- 7499815 TI - Extended-spectrum beta-lactamase in Salmonella spp. PMID- 7499814 TI - Bacterial contamination of ventilators. PMID- 7499816 TI - Guidelines on the control of methicillin-resistant Staphylococcus aureus in the community. Report of a combined Working Party of the British Society for Antimicrobial Chemotherapy and the Hospital Infection Society. PMID- 7499817 TI - The significance of nasal carriage of Staphylococcus aureus and the incidence of postoperative wound infection. AB - Staphylococcus aureus infections are associated with considerable morbidity and, in certain situations, mortality. The association between the nasal carriage of S. aureus and subsequent infection has been comprehensively established in a variety of clinical settings, in particular, patients undergoing haemodialysis and continuous ambulatory peritoneal dialysis (CAPD), and in patients undergoing surgery. Postoperative wound infections are associated with a high degree of morbidity and represent an important medical issue. Until recently, eradication of S. aureus nasal carriage by various topical and systemic agents had proved unsuccessful. Mupirocin is a novel topical antibiotic with excellent antibacterial activity against staphylococci. Recent studies have demonstrated that intranasal administration of mupirocin is effective in eradicating the nasal carriage of S. aureus and in reducing the incidence of S. aureus infections in haemodialysis and CAPD patients. It has been suggested that sufficient evidence now exists to test the hypothesis that eradication of the carrier state in surgical patients preoperatively may reduce the incidence of S. aureus postoperative wound infections. PMID- 7499818 TI - Disinfection and sterilization practices in Mexico. AB - We evaluated antisepsis, disinfection, and sterilization procedures at 22 hospitals in the state of Yucatan, Mexico, which provide services for approximately 80% of the population. The percentage of hospitals that followed standard recommendations for diverse antisepsis procedures were as follows: surgical scrub, 41%; surgical site preparation, 68%; central intravenous (iv) catheters, 61%; peripheral iv catheters, 86%; urinary catheters, 41%, and umbilical cord care, 5%. Inappropriate procedures typically involved the use of benzalkonium chloride and mercury compounds. Adequate sterilization procedures were observed for sharp surgical instruments in 9%, for blunt surgical instruments in 81%; for linen in 90%; for surgical brushes in 9%; for metal prostheses in 100%; for plastic prostheses in 57%; and for disinfection of endoscopes in 30%. The most common errors were the use of too short an exposure time in steam sterilizers or dry heat sterilizers, and the use of benzalkonium chloride as a sterilizing agent. Only 14% of hospitals used commercial spore preparations for autoclave monitoring. For the reuse of disposables, 50-94% of hospitals employed inappropriate sterilization or low-level disinfection procedures. Similar conditions are likely to exist in hospitals throughout the country. The establishment of strict regulations and hospital training programmes for disinfection and sterilization procedures, and the reuse of disposable devices is urgently needed in Mexico. PMID- 7499819 TI - Genomic fingerprinting Acinetobacter baumannii: amplification of multiple inter repetitive extragenic palindromic sequences. AB - Acinetobacter species are important nosocomial pathogens. A rapid and sensitive identification system, capable of providing strain identity at the genetic level, is required to identify outbreak strains and facilitate the early implementation of infection control procedures. Repetitive extragenic palindromic (REP) elements, have been identified in numerous bacteria and these genomic sequences provide useful targets for DNA amplification. A method for amplifying inter-REP DNA sequences, REP-multiple arbitrary amplicon profiling (REP-MAAP), is described and applied to 29 Acinetobacter baumannii from clinical samples. Amplified polymorphic DNA patterns were demonstrated for all isolates and those displaying identical REP-MAAP patterns were considered identical at the genetic level. In the spring of 1993, 10 intensive care unit patients had endotracheal colonization with A. baumannii (five with REP-MAAP I and five with REP-MAAP II patterns). These findings suggested nosocomial transmission of organisms which was terminated by standard infection control measures. No further A. baumannii were detected until the winter of 1993 when isolates of different REP-MAAP groups emerged, suggesting that factors other than nosocomial transmission were implicated. PMID- 7499820 TI - Outbreak caused by two multi-resistant Acinetobacter baumannii clones in a burns unit: emergence of resistance to imipenem. AB - Since early 1992 an increased number of tobramycin- and imipenem-resistant Acinetobacter spp. were observed causing colonization, wound infections, and bacteraemias in a burns and plastic surgery unit. This raised the question of whether this outbreak was caused by a single or by multiple Acinetobacter spp. clones. To study this, 97 Acinetobacter spp. isolates from clinical samples from different hospital units as well as isolates from the environment and the hands of the staff were characterized by antibiogram, plasmid profile and ribotyping. Two dominant multi-resistant A. baumannii clones were identified; one of them was sensitive to polymyxin B only. There was a close correlation between the results obtained by plasmid profiling and ribotyping. No common environmental source or significant hand carriage, or spread of these strains outside the unit were detected. The burns patients were the most likely reservoir, and strain transmission occurred in spite of strict control measures. PMID- 7499822 TI - DNA fingerprinting of Pseudomonas aeruginosa serotype O11 by enterobacterial repetitive intergenic consensus-polymerase chain reaction and pulsed-field gel electrophoresis. AB - We report the use of pulsed-field gel electrophoresis (PFGE) and enterobacterial repetitive intergenic consensus (ERIC)-based polymerase chain reaction (PCR) to characterize clinical isolates of Pseudomonas aeruginosa serotype O11 collected from an incident of hospital-acquired infection. Both typing techniques differentiated 20 different strain types among seven epidemiologically related isolates and 22 epidemiologically unrelated isolates. There was complete concordance between these two techniques. Our results indicate that the ERIC based PCR technique represents a rapid and simple means for typing P. aeruginosa serotype O11 with a level of discrimination equivalent to that of PFGE. PMID- 7499821 TI - Non-enteritic aeromonas infections in hospitalized patients. AB - Aeromonas species were isolated from specimens other than faeces from 59 hospital inpatients over a 15 year period. Of the isolates, 79.7% were regarded as clinically significant, with skin and soft tissues and blood cultures as the commonest sites of infection. Of the isolates, 52.5% were hospital-acquired, and 55.9% of patients had serious underlying disease. Community-acquired infections in previously healthy individuals accounted for only 13.6% of isolates. However unlike many other opportunistic infections, aeromonas infection was not closely associated with prior antibiotic therapy, nor was there a significant increase in the frequency of infection over the study period. PMID- 7499823 TI - Utility of surveillance bacterial cultures in neonatal exchange blood transfusions. AB - Invasive procedures, like exchange blood transfusions via the umbilical vein, potentially expose the neonate to nosocomial infection. Attempts to limit hospital-acquired infection following exchange transfusions (ETF) in our unit have included umbilical vein blood cultures and swabs from umbilical stumps. The value of this surveillance was examined. Forty-four neonates undergoing ETF were studied prospectively. Specimens for bacterial cultures were taken from the umbilical stump and umbilical vein immediately before and after ETF and results correlated with clinical outcome and antibiotic use. Except for staphylococci, bacteria cultured in the asymptomatic neonates were similar to those cultured from neonates who had signs of infection. Polymicrobial cultures were obtained from both umbilical vein blood and stump specimens suggesting contamination with colonizing organisms. Surveillance bacterial culture results did not influence antimicrobial therapy. Therefore, we advocate microbiological investigations only when clinical infection is suspected. PMID- 7499824 TI - Surveillance cultures on neonatal intensive care units. PMID- 7499825 TI - Statistical analysis of results. PMID- 7499826 TI - Guidelines for good practice in central venous catheterization. PMID- 7499827 TI - Cross-linking of ICAM-1 on T cells induces transient tyrosine phosphorylation and inactivation of cdc2 kinase. AB - Intercellular adhesion molecules (ICAM)-1 and -3 coexist on T lymphocytes and are counter-receptors for the integrin LFA-1. Signaling through ICAM-3 stimulates a number of T cell functions and involves phosphorylation of Fyn, Lck, CD45, and other proteins. In contrast, this type of specific signaling event has not been described for signaling through ICAM-1. Here, tyrosine phosphorylation of cellular proteins was examined after cross-linking of ICAM-1. Tyrosine phosphorylation of the 34-kDa cdc2 protein kinase was induced transiently after stimulation of the leukemic T cell line, Molt-3, or peripheral blood T cells. Stimulation through ICAM-1 had no effect on constitutive presence of cdc2 or phosphorylation of cdc2 on threonine. cdc2 kinase activity was constitutive in peripheral blood T cells, and transient inhibition of kinase activity after ICAM 1 stimulation correlated kinetically with phosphorylation of cdc2 on tyrosine. PMID- 7499828 TI - Human IL-17: a novel cytokine derived from T cells. AB - A cDNA encoding human IL-17 (hIL-17) was cloned from a CD4+ T cell library. The predicted 155-amino acids sequence contains an N-terminal signal peptide and exhibits 72% amino acid identity with HVS13, an open reading frame from a T lymphotropic Herpesvirus saimiri, and 63% with murine CTLA8. High levels of hIL 17 were induced from primary peripheral blood CD4+ T cells upon stimulation. When expressed in CV1/EBNA cells, recombinant hIL-17 was secreted in both glycosylated and nonglycosylated forms. A hIL-17.Fc fusion protein and supernatants from cells transfected with hIL-17 induced IL-6 and IL-8 production and enhanced the surface expression of the intracellular adhesion molecule-1 (ICAM-1) in human fibroblasts. PMID- 7499829 TI - Differential expression of alternate mB7-2 transcripts. AB - The murine B7-2 (mB7-2) costimulatory molecule is expressed on APCs early during the course of an immune response, suggesting that it may play a pivotal role in the decision between T cell activation and anergy. Murine B7-2 mRNA displays a restricted pattern of expression; it is inducible in B cells, T cells, NK cells, and dendritic cells, but constitutively expressed in unstimulated monocytes. The constitutive and inducible expression of mB7-2 on distinct cell types indicates that mB7-2 is regulated differentially. To further characterize mB7-2 transcripts, we employed 5' rapid amplification of cDNA ends and reverse transcriptase-PCR to examine transcripts expressed in a variety of types of APCs and analyzed the genomic organization of the mB7-2 gene. We report here that the mB7-2 locus consists of 12 exons and demonstrate that exons 1 through 5 can be used in alternative fashions to produce five distinct transcripts, differing in their 5' untranslated and signal regions. The expression of these transcripts differs in distinct types of APCs and is modulated by stimuli that activate B cells. These results demonstrate that mB7-2 transcripts are differentially regulated in a tissue-specific fashion and in response to activation stimuli. PMID- 7499830 TI - Glycosylphosphatidylinositol-modified murine B7-1 and B7-2 retain costimulator function. AB - Glycosylphosphatidylinositol (GPI)-modified variants of murine B7-1 and B7-2 cell surface costimulators were produced via chimerization with alternative GPI modification signal sequences from decay-accelerating factor (DAF). GPI anchorage was verified by demonstrating phosphatidylinositol-specific phospholipase C (PI PLC) sensitivity of the chimeric polypeptides in both immunofluorescence/flow cytometric and immunoprecipitation analyses. The various GPI-modified chimeric B7 1:DAF and B7-2:DAF polypeptides were shown to retain costimulator function, in both an in vitro proliferation assay and an in vivo triggering of cytotoxicity assay. The findings indicate that costimulator function for both B7-1 and B7-2 is not dependent upon native hydrophobic transmembrane anchorage. Moreover, the functionality of the GPI-modified variants in enhancing the immunogenicity of the murine T lymphoma line EL-4 suggests a novel route for generating APC-centered immunotherapeutics, including cellular cancer vaccines, that is based upon protein transfer of GPI-modified costimulators. PMID- 7499831 TI - Alloantigen-induced human lymphocytes rendered nonresponsive by a combination of anti-CD80 monoclonal antibodies and cyclosporin-A suppress mixed lymphocyte reaction in vitro. AB - Induction of a state of long-term, alloantigen-specific T cell nonresponsiveness has significant implications for human transplantation. It has been previously described that alloantigen-specific anergy may be induced by addition of cyclosporin-A together with anti-CD80(B7-1) mAb to a MLR. In this study we endeavored to verify whether alloantigen-induced PBL rendered anergic by the addition of a combination of anti-B7 mAb and cyclosporin-A during a MLR had a suppressive effect when added to autologous lymphocytes activated in MLR. We found that: 1) the addition of cells rendered anergic by this procedure to a MLR suppress both proliferative and cytotoxic response of autologous responsive PBL to either the same or third-party stimulator cells; 2) the suppressive effect is limited to alloantigen-induced T cell activation, as addition of anergic cells does not influence mitogen- or antigen-induced proliferation of autologous responsive T cells; 3) nonresponsiveness of suppressed cells cannot be reversed by either subsequent restimulation with allogeneic cells or addition of exogenous IL-2 to the cultures; 4) the suppressive effect is apparently not due to secretion of anergic cell-derived soluble factors, but it seems to be dependent on cell-cell contact between anergic, responsive, and stimulator cells. These data suggest that: 1) the delivery of a direct signal mediated by anergic lymphocytes through a cell-cell contact is likely to be the mechanism responsible for the suppressive effect here described; 2) anergic cells may propagate alloantigen-specific tolerance to potentially responsive autologous lymphocytes. Preliminary experiments indicate that anti-CD86(B7-2) mAb may play a similar role in the generation of alloantigen-induced nonresponsiveness. PMID- 7499832 TI - Natural killer cell ontogeny in the athymic rat. Relationship between functional maturation and acquired resistance to E1A oncogene-expressing sarcoma cells. AB - Newborn nude rats are NK cell deficient and highly susceptible to tumor induction by NK-susceptible, E1A oncogene-expressing, BHK-21 sarcoma cells, whereas adults have normal NK activity and are tumor resistant. This model of NK cell-related tumor rejection was used to characterize age-related functional maturation of NK cells during nude rat ontogeny. Small numbers of cytolytic incompetent, NKR-P1+ cells were detected in the spleens of newborns. During the first month of life, there was an age-related increase in the numbers of splenic NK cells, followed by a stepwise maturation in their cytolytic competence. Three functional stages in NK cell cytolytic activity were apparent. A nonlytic newborn stage was followed by a transitional stage, during which absent or relatively weak spontaneous cytolytic activity was augmented by prolonged NK cell exposure to susceptible target cells. In the final stage, both spontaneous and augmented cytolytic activity increased toward the adult level that was attained by 4 wk of age. The data suggest the existence of a transient stage in NK cell ontogeny when triggering through the NKR-P1 molecule is functionally immature, and confirm the importance of a cytolytic competent, NK cell response for nude rat rejection of E1A oncogene-expressing tumor cells. PMID- 7499833 TI - Tumor-associated E6 protein of human papillomavirus type 16 contains an unusual H 2Kb-restricted cytotoxic T cell epitope. AB - We previously showed that CTL from H-2b mice immunized against the E6 protein of human papilloma virus (HPV) type 16 recognized a 10-mer peptide corresponding to amino acids 131 to 140. We show in this study that the minimal epitope, 130 to 137, is a 8-mer peptide presented by H-2Kb class I molecules. At position P8, the 130 to 137 peptide contains a hydrophobic methionine anchor residue, but P3 and P5 do not contain the typical anchor residues that are frequently found in Kb bound peptides. Analysis with alanine-substituted peptides indicates that the tryptophan at P3 acts as an alternative anchor mediating Kb binding, while an arginine at P2 is a TCR contact residue. Synthetic 9-mer peptides corresponding to residues 130 to 138 are as efficiently recognized by CTL as 130 to 137 peptides. Analysis of extracts of E6-expressing cells suggests that Ag processing may produce multiple peptides containing the minimal 130 to 137 epitope. In vitro binding studies indicate that Kb binding of peptide 130 to 137 is approximately five orders of magnitude less efficient than Kb binding of previously identified CTL epitopes. In contrast, the E6 protein contains another potential CTL epitope in the region of amino acids 41 to 50. A synthetic peptide spanning this region binds very strongly to Kb and is capable of stimulating a strong peptide-specific CTL response. In the context of the whole protein, however, this epitope remains cryptic. PMID- 7499834 TI - CD40 signaling-mediated induction of Bcl-XL, Cdk4, and Cdk6. Implication of their cooperation in selective B cell growth. AB - Signals sent through CD40 play crucial roles in B cell differentiation, including blocking apoptosis of germinal center B cells. In this study, using a murine B cell WEHI-231 line that undergoes apoptosis by the cross-linking of surface Ag receptors (sIgM), we have demonstrated that CD40 signalings are linked to induction of the Bcl-xL, Cdk4, and Cdk6 proteins whose expression was significantly suppressed by the apoptotic signal through sIgM. Mutational analyses of CD40 revealed that the domain of human CD40 required for blocking apoptosis of WEHI-231 cells coincides with that required for Bcl-xL induction. Signals through sIgM arrest cells in the G1 phase of the cell cycle, which is followed by apoptosis. However, while constitutive expression of Bcl-XL leads to the inhibition of apoptosis. Nevertheless, Bcl-xL fails to induce S phase entry. By CD40 signalings, both Cdk4 and Cdk6 resume their normal expression levels, which are sufficient for passing the restriction point in G1 even in the presence of the apoptotic signals mediated by sIgM. These results suggest that cooperation of Bcl-xL, Cdk4, and Cdk6 induced by CD40 signaling plays a key role in CD40 mediated selective growth of B cells. PMID- 7499835 TI - Surrogate light chain-dependent selection of Ig heavy chain V regions. AB - Bone marrow B cell precursors frequently rearrange the Ig heavy chain variable (VH) gene segment VH81X. It is puzzling, therefore, that mature B cells in adult mice rarely express mu-heavy chains bearing this VH gene segment. We show in this work in transformed pre-B cell lines that two VH81X/mu-chains that differ in their VH-D-JH joining sequences are not assembled covalently with the B cell precursor-specific surrogate light (SL) chain and are not expressed on the cell surface. From these findings, we propose that a B cell precursor clonally expands and proceeds to the next developmental stage only if it expresses a mu-chain with a VH domain that, together with the SL chain, directs the formation of a signal transducing mu/SL chain membrane complex. Therefore, a checkpoint exists early in B cell development, at which SL chain not only screens B cell precursors for the presence of a full-length mu-chain, but also for a VH domain that promotes the assembly of a mu/SL chain complex. PMID- 7499836 TI - Cross-linking of the Fas/APO-1 antigen suppresses the CD3-mediated signal transduction events in human T lymphocytes. AB - The Fas/APO-1 Ag, a member of the TNF-R family, mediates apoptosis (programmed cell death, PCD). PCD has been recognized to be important in the regulation of normal and autoimmune responses. However, the underlying molecular mechanisms are not fully known. To elucidate the role of the Fas Ag in lymphocyte activation, we investigated the influence of Fas Ag cross-linking on CD3/TCR-mediated signal transduction in IL-2-dependent human T cell lines. Early signal transduction events were evaluated in 10-day-old T cell lines using anti-Fas and anti-CD3 mAbs. Incubation of Indo-1-loaded cells with anti-Fas mAb (10 micrograms/ml), but not with an isotypic control mAb almost completely inhibited the anti-CD3 mediated Ca2+ influx. This inhibition was mAb concentration dependent and was detected after incubation for at least 15 min. Cross-linking of the Fas Ag did not affect the anti-CD3-mediated release of Ca2+ from the internal stores or the thapsigargin-mediated Ca2+ influx. Anti-Fas mAb-incubated cells also showed decreased CD3-mediated inositol 1,4,5-trisphosphate production. Preincubation of cells with anti-Fas mAb inhibited the anti-CD3-mediated tyrosine phosphorylation of multiple cellular proteins. Cross-linking of the Fas Ag for up to 6 h did not alter the expression of CD3 on the surface of the T cells. Preincubation of Indo 1-loaded EBV-transformed B cells with anti-Fas mAb did not affect the anti-mu Ab induced increase in intracellular calcium ([Ca2+]i). Our results show that cross linking of the Fas Ag can suppress the receptor-mediated signaling process in human T but not B cells. PMID- 7499837 TI - Expression of sialyl Lewis(x) and Lewis(x) defines distinct stages of chicken B cell maturation. AB - Commitment of cells to the B lineage in chickens occurs only during a brief period of embryogenesis. B cell progenitors then progress through discrete developmental stages resulting in the production of mature B cells that are competent to form a functioning humoral immune system in the adult bird. During embryogenesis, chicken B cell precursors undergo tissue and developmental stage specific changes in cell-surface glycosylation. Immature B cell progenitors that migrate to the bursa of Fabricius express the carbohydrate epitope sialyl Lewis(x). Such cells undergo initial clonal expansion within the bursa without undergoing gene conversion. Beginning between days 15 and 17 of embryogenesis, B cells in the bursa undergo a tissue specific change in surface glycosylation that results in the loss of sialyl Lewis(x) expression and the acquisition of the related carbohydrate structure Lewis(x). Expression of Lewis(x) identifies B lymphocytes that have begun to undergo gene conversion. Before emigration from the bursa, bursal lymphocytes undergo another phenotypic switch in glycosylation and down-regulate Lewis(x) expression. Therefore, developmental switches in glycosylation allow us to distinguish three populations of B cells in the bursa of Fabricius at defined stages of development and suggest that regulation of cell surface glycosylation plays a role in B cell development in the chicken. PMID- 7499838 TI - Proteoglycan form of colony-stimulating factor-1 (proteoglycan-100). Stimulation of activity by glycosaminoglycan removal and proteolytic processing. AB - A proteoglycan had been isolated from the conditioned media of a human osteosarcoma cell line and had tentatively been named proteoglycan-100 (PG-100) because of the size of its core glycoprotein. Amino acid sequencing of the purified proteoglycan and cDNA analysis were consistent with the assumption that PG-100 is identical with the proteoglycan form of CSF-1 (or macrophage colony stimulating factor). PG-100 induced mouse macrophage differentiation. Proliferation of macrophages was stimulated in a dose-dependent manner. On a molar basis, however, about 100- to 300-fold higher doses of PG-100 than of recombinant human (rh)CSF-1 were required for the half-maximal growth-stimulating effect. Upon enzymatic removal of the glycosaminoglycan chain, the purified core protein exhibited higher activity, but was still about 20-fold less active than rhCSF-1. Incubation of the purified proteoglycan for 48 h at 37 degrees C led to the formation of a glycosaminoglycan-free 50-kDa fragment either by autoproteolysis or by the action of a protease not yet identified. The purified fragment exhibited almost the same biologic activity as rhCSF-1. The glycosaminoglycan chain of the growth factor was not only shown to inhibit CSF-1 activity but also to increase the stability of the core protein when the CSF-1 producing osteosarcoma cells were maintained in a collagen lattice. These findings provide a link between a soluble, highly active cytokine and its extracellular matrix storage form of comparatively low activity. PMID- 7499839 TI - Sex hormone and IL-6 regulation of antigen presentation in the female reproductive tract mucosal tissues. AB - The purpose of this study was to determine whether Ag presentation takes place in the female reproductive tract and if the stage of the reproductive cycle, sex hormones, and IL-6 regulate uterine Ag presentation. Purified epithelial and mixed stromal cells were incubated with sensitized lymph node T lymphocytes in the presence of OVA. Ag presentation was highest at proestrus, the stage of the reproductive cycle that immediately precedes ovulation when estradiol levels peak, and lowest at estrus, when sperm enter the uterus in preparation for fertilization. Further, these studies demonstrate that estradiol given to ovariectomized rats increases uterine cell Ag presentation relative to that in controls. In the presence of mouse anti-rat Ia Ab, Ag presentation was blocked, indicating that Ag presentation by uterine cells is MHC class II restricted. In studies to analyze Ag presentation at sites distal to the female reproductive tract, we found that Ag presentation by adherent cells from the spleen is elevated at proestrus relative to that at other stages of the estrous cycle. When IL-6 was placed in the uterine lumen of ovariectomized rats, Ag presentation by epithelial and mixed stromal cells as well as mitogenesis by spleen cells were increased significantly relative to those in controls. These studies demonstrate that the female reproductive tract is an inductive site for immune responses and that mucosal immune protection may be either enhanced or suppressed depending on the endocrine balance when the female reproductive tract is exposed to pathogens. PMID- 7499840 TI - Modulation of TCR-mediated signaling pathway by thymic shared antigen-1 (TSA 1)/stem cell antigen-2 (Sca-2). AB - Thymic shared antigen-1 (TSA-1) is a glycosyl-phosphatidylinositol (GPI)-anchored differentiation Ag expressed on murine lymphocytes, and is identical to stem cell Ag-2 (Sca-2). Using newly established mAb against TSA-1/Sca-2, we have previously shown that surface TSA-1 expression is induced upon activation in T cells, and that anti-TSA-1 inhibits IL-2 production induced by anti-CD3 stimulation in T cell hybridomas. In the present study, we have analyzed the functional role of TSA-1 during T cell activation using normal T cells, T cell hybridomas, and transfected Jurkat cell lines that expressed either GPI-anchored or transmembrane form of TSA-1. Anti-TSA-1 inhibited IL-2 production from normal T cells stimulated with soluble anti-CD3 plus accessory cells. Anti-TSA-1 exhibited the inhibitory effect on T cells, but not on accessory cells, because anti-TSA-1 inhibited IL-2 production in Jurkat cells transfected with TSA-1 cDNA, but not in control transfectant. A transmembrane form of TSA-1 was expressed in Jurkat cells by fusing the extracellular portion of TSA-1 to the transmembrane and cytoplasmic regions of the class 1 Db. The analysis using this transfectant revealed that anti-TSA-1-mediated inhibition of IL-2 production did not require the GPI anchor of TSA-1. Finally, in addition to the inhibition of IL-2 production, tyrosine phosphorylation of CD3 zeta-chains observed following TCR stimulation, one of the important early activation events, was markedly reduced by anti-TSA-1. These results imply that TSA-1/Sca-2 plays an important regulatory role in the TCR signaling pathway of activated T cells in addition to its role in T cell differentiation. PMID- 7499842 TI - Breaking immunologic ignorance to an antigenic peptide of simian virus 40 large T antigen. AB - Expression of antigenic proteins in the periphery may result in immunologic tolerance, immunologic ignorance, or autoimmunity. It is not clear why some Ags induce tolerance, whereas others activate Ag-reactive T cells. The physical nature of the Ag, the developmental timing of Ag expression, the priming of reactive lymphocytes, and the level of Ag expression are possible factors determining the immunologic response to extra-thymic Ags. Expression of the whole SV40 large T Ag (SV40-T) induces transformation of T antigen-expressing cells in vivo, and this phenomenon has been postulated to be the triggering event that leads to autoimmunity in some transgenic mouse models. Here we present a model in which a nononcogenic, yet antigenic, fragment of the SV40-T (SV40-Tfrag) is expressed specifically in pancreatic islet beta-cells. In contrast to whole SV40 T transgenic mice, SV40-Tfrag mice that are also transgenic for a TCR specific for the SV40-T are ignorant of Ag in vivo. They do not respond in vivo to the tissue-specific SV40-Tfrag Ag even after priming, but are fully responsive in vitro. This immunologic ignorance cannot be broken after activated SV40-T reactive T cells are transferred into sublethally irradiated mice expressing the islet-specific SV40-Tfrag. However, similar adoptive transfer experiments in mice co-expressing B7-1 and SV40-Tfrag on islet cells specifically lead to Ag recognition and diabetes. This demonstrates that in some circumstances the presence of (primed) reactive T cells is not sufficient to break tolerance; rather, costimulation is additionally required to elicit an autoimmune response. This also suggests that SV40-T-induced cellular transformation is important for the autoimmune response directed against SV40-T in other tissue-specific transgenic models. PMID- 7499841 TI - Bacterial lipoproteins may substitute for cytokines in the humoral immune response to T cell-independent type II antigens. AB - Bacterial lipoproteins share a common structural motif that has been shown to stimulate proliferation and Ig secretion of murine B cells, in a manner distinct from that mediated by LPSs. Studies of lipoprotein-mediated B cell activation utilized heterogeneous populations of lymphoid cells, leaving unresolved their ability to directly activate resting B cells, as well as their ability to interact with other B cell stimuli. Using highly enriched and/or sort-purified resting murine B cells, we demonstrate that, in contrast to previous reports, lipoproteins (lipoprotein-D, lipoprotein-OspA, and/or the synthetic analogue Pam3Cys) stimulate little, if any, proliferation or Ig secretion in resting B cells. However, when combined with a multivalent membrane (m)Ig-mediated cross linking signal, dextran-conjugated anti-IgD Abs (alpha delta-dex), lipoproteins mediate up to 10,000-fold inductions in IgM secretion and up to 25-fold enhancements in cellular proliferation relative to that observed with alpha delta dex alone, in the absence of added cytokines. This mIg-mediated enhancement of Ig secretion was not observed when B cells were stimulated with bivalent, unconjugated anti-Ig. CD40 ligand (CD40L), shows a similar, although somewhat more moderate, synergy with lipoproteins for induction of proliferation and IgM secretion. By contrast, lipoproteins by themselves are relatively ineffective at costimulating Ig secretion in the presence of various combinations of cytokines. These data suggest that bacteria may induce Ag-specific humoral immunity through the action of bacterial polysaccharides that mediate an Ag-specific multivalent mIg signal, in concert with bacterial lipoproteins that deliver ancillary signals, without a requirement for recruitment of non-B cell types. PMID- 7499843 TI - Suppressor T cell-activating macrophages in ultraviolet-irradiated human skin induce a novel, TGF-beta-dependent form of T cell activation characterized by deficient IL-2r alpha expression. AB - Because UV-induced epidermal macrophages (UV-Mph) preferentially activate CD4+ T suppressor-inducer cells and induce tolerance, we hypothesized that they differentially up-regulate T cell early activation genes compared with constitutive epidermal APC, Langerhans cells. We used epidermal cells from UV exposed (UV-EC) and control (C-EC) human skin to stimulate allogeneic CD4+ T lymphocytes. Reverse transcriptase-PCR revealed that both C-EC (Langerhans cells) and UV-EC (UV-Mph) induced 10(3)- to 10(6)-fold increases in IL-2 mRNA. However, while T cells stimulated by C-EC for 48 h showed a greater than 10(3)-fold increase in IL-2R alpha mRNA, those stimulated by UV-EC did not (n = 5, p = 0.004). Flow cytometry demonstrated that 4.1 +/- 2.3% of unstimulated CD4+ lymphocytes expressed cell surface IL-2R alpha, which increased to 15.7 +/- 1.8% upon stimulation by C-EC for 48 h, but stimulation by UV-EC failed to increase the IL-2R alpha+ population (n = 3, p = 0.038). The addition of neutralizing anti TGF-beta Abs to UV-EC-stimulated cultures restored CD4+ cell surface IL-2R alpha expression to 12.9 +/- 0.2%. CD4+ T cell activation by UV-Mph is distinct from previously described models of tolerance such as Th2 activation (IFN-gamma mRNA was induced and IL-4 mRNA was not) and Th1 anergy (IL-2 mRNA levels induced by UV EC and C-EC were similar). Furthermore, costimulatory signals were provided by UV Mph; CTLA4-Ig and LFA-3-Ig fusion proteins and Abs to CD2, LFA-3, LFA-1, and ICAM 1 inhibited UV-Mph-induced T cell proliferation. Thus, the altered immune outcome induced by UV-Mph (tolerization) compared with Langerhans cells (sensitization) is reflected as a novel mechanism of initial CD4+ T cell early activation gene expression characterized by TGF-beta-dependent deficient IL-2R alpha expression. PMID- 7499844 TI - Durable elimination of high affinity, T cell-dependent antibodies by low molecular weight antigen arrays in vivo. AB - Ongoing Ab responses to a T cell-dependent Ag can be suppressed in hyperimmune animals by exogenous, multivalent Ag arrays. The pharmacologic basis for this suppression was studied by varying the molecular mass, ligand valence, and dose of Ag arrays, and then determining their efficacy, pharmacokinetics, and tissue distribution. Arrays ranging in molecular mass from 30 to 500 kDa caused initial clearance of specific serum Abs, but only the smaller arrays caused persistent suppression despite their relatively lower binding avidity and shorter retention in vivo. Suppression by the smaller arrays at lower doses was biphasic, implying two distinct modes of Ab elimination. High affinity IgG was eliminated preferentially, as shown by calibrated variable ligand-density ELISA. Suppressive arrays were localized discretely in the splenic germinal centers of hyperimmune animals. These results indicate that Ag array mass, ligand valence, and dose all play critical roles, and histologic compartmentalization may also be a pertinent parameter, in determining suppressive efficacy in vivo. PMID- 7499846 TI - In vivo association between p56lck and MAP kinase during IL-2-mediated lymphocyte proliferation. AB - We previously reported that p56lck expression is upregulated in human B lymphocytes upon mitogenic stimulation. In this report, we characterized the molecules associated with p56lck in vivo in leukemic B cells costimulated with anti-mu Ab and IL-2 for 72 h. In vitro phosphorylation after p56lck immunoprecipitation indicated that p56lck is associated in vivo with the beta chain of the IL-2 receptor and p42 MAP kinase as well as a number of other proteins. Moreover, p56lck-associated MAP kinase is tyrosine and threonine phosphorylated, suggesting that it is activated. Prevention of DNA synthesis with aphidicolin abrogated this molecular association, and furthermore, cell cycle analysis with IL-2-dependent T cells showed that in cells in G1, MAP kinase was not associated to p56lck, whereas this p56lck-MAP kinase association was observed when cells are in S phase. Thus, p56lck and MAP kinase are only associated during S phase. These data suggest that MAP kinase in association with p56lck is directly involved in the control of IL-2-mediated DNA synthesis of both B and T lymphocytes. PMID- 7499845 TI - Identification of IFN-gamma receptor binding sites for JAK2 and enhancement of binding by IFN-gamma and its C-terminal peptide IFN-gamma(95-133). AB - The tyrosine kinase JAK2 is an integral part of the signal transduction pathways of a number of cytokines and growth factors, including IFN-gamma. Previously, we identified a species-nonspecific binding site for the C terminus of IFN-gamma, encompassed by IFN-gamma peptide IFN-gamma(95-133), on the membrane proximal region of the cytoplasmic domain of the IFN-gamma R alpha-chain. Using both a radioligand binding assay and coimmunoprecipitation with antireceptor antiserum, we were able to demonstrate specific interaction of JAK2 with the murine IFN gamma R(MIR) alpha-chain. Furthermore, this interaction is increased by the addition of murine IFN-gamma or its C-terminal peptide, muIFN-gamma(95-133). We also identified two regions of the cytoplasmic domain of the receptor that interact with JAK2 using synthetic peptides of the MIR alpha-chain in receptor competition studies. These regions are encompassed by receptor peptide MIR(283 309), which is adjacent to the membrane proximal region at which the C terminus of IFN-gamma binds, and receptor peptide MIR(404-432), which lies near the C terminus of the receptor, encompassing a potentially important phosphorylation site. These data show site-specific interaction between JAK2 and IFN-gamma with the IFN-gamma R and have broader implications for the role of the IFN-gamma ligand in the IFN-gamma signal transduction pathway. Furthermore, the data support previous studies that demonstrated that intracellular IFN-gamma plays a role in cell activation. PMID- 7499847 TI - Reconstitution ratio is critical for alloreactive T cell deletion and skin graft survival in mixed bone marrow chimeras. AB - Although allogeneic bone marrow (BM) transplantation is an effective way to induce donor-specific tolerance, its clinical application is hampered because of the risk associated with vigorous myeloablative pretransplant conditioning. One approach to overcome this problem is to establish a lower chimeric state by mild myeloablation. It is not clear, however, whether there is a threshold in the extent of chimerism that is required for tolerance induction. In this study, we establish a mixed BM chimera system to examine the correlation between the reconstitution ratio of BM chimerism, donor-reactive T cell deletion, and skin graft acceptance using I-E alpha transgenic C57BL/6 mice as the BM and skin graft donors and Ly5 congenic C57BL/6 mice as the recipients. In this system, the class II MHC molecule I-E was the transplantation Ag, and the extent of I-E-reactive T cell deletion was determined by flow cytometry using a mAb specific for the V beta 11 TCR. The degree of BM chimerism was measured by examining the expression of donor-derived Ly5.2 and host-derived Ly5.1 on peripheral blood cells. Transplantation of I-E+ transgenic donor BM cells resulted in deletion of V beta 11+CD4+ T cells in recipient's PBL, and the extent of deletion was proportional to the degree of chimerism. When mice of different degrees of chimerism were tested for skin graft survival, we found that recipient mice with > 30% chimerism could accept skin grafts from I-E+ donor mice, whereas those with < 10% chimerism showed prolonged but not permanent graft survival. These findings revealed the sequence of events for induction of tolerance. First, the degree of BM chimerism determines the number of I-E+ cells in the thymus, which then elicits negative selection of I-E-reactive T cells in a form of clonal deletion. The extent of T cell deletion ultimately determines the mode of tolerance. These data provide experimental evidence for the potential use of partial chimerism by bone marrow transplantation for the induction of donor-specific tolerance in clinical settings. PMID- 7499848 TI - Induction of germ-line gamma 1 and epsilon Ig gene expression in murine B cells. IL-4 and the CD40 ligand-CD40 interaction provide distinct but synergistic signals. AB - The interaction between B cell CD40 and its ligand (CD40L) on activated Th cells provides a critical signal necessary for T cell-dependent isotype switching. Previous studies suggest that this signal might be important in regulating isotype switching at the level of germ-line Ig transcription. To assess the effects of the CD40L-CD40 interaction on germ-line Ig transcript expression in murine B cells, a membrane-bound form of mouse CD40L was expressed in the baculovirus system. We show that stimulation of resting splenic B cells with CD40L-expressing Sf9 cells induces germ-line gamma 1 and epsilon transcripts independently of cytokines. The CD40-mediated induction cannot be blocked by anti IL-4 Ab and is not mediated by other cytokines secreted endogenously in response to CD40 stimulation. Importantly, stimulation with CD40L and IL-4 together has a significant synergistic effect on germ-line transcript expression. Stimulation of CD40 does not activate the NF-IL-4-gamma 1 DNA binding factor believed to be required for IL-4-dependent germ-line gamma 1 transcription. Moreover, mutation of the NF-IL-4-gamma 1 DNA binding site in a germ-line gamma 1 promoter luciferase reporter gene construct completely ablates IL-4 responsiveness but has no effect on responsiveness to CD40L in transient transfection assays. These results demonstrate that the CD40L-CD40 interaction and IL-4 activate germ-line Ig gene transcription by distinct but synergistic mechanisms and suggest that multiple signals may be required to induce sufficient germ-line transcription and/or germ-line transcript levels necessary to target switch recombination. PMID- 7499849 TI - Evaluation of loss and change of specificity resulting from random mutagenesis of an antibody VH region. AB - Most data available from in vivo sources regarding the impact of somatic hypermutation on Ab V region structure and function are heavily biased due to the influence of clonal selection. In an effort to address this issue directly, we "randomly" introduced point mutations throughout the length of the VH region of an anti-p-azophenylarsonate (Ars) Ab expressed as an Fab in the phage display format. This was accomplished by means of an error-prone PCR with two protocols, which resulted in two mutant libraries. The nature of the nucleotide substitutions obtained from each protocol differed from each other and resulted in different frequencies of phage clones that did not appear to contain Fab on their surfaces. However, the majority of mutants in both libraries lacked detectable Fab expression. Screening of the library containing the most expressed Fabs for those that had gained affinity for structurally related haptens yielded two independent mutants that lacked detectable affinity for Ars and had high affinity for p-azophenylsulfonate. These mutants both contained amino acid substitutions from Asn to Ser or Thr at VH CDR1 position 35, a putative Ars contact residue. In this paper, we discuss the significance of these data with regard to the frequencies of V region loss of function, gain of increased affinity, and gain of altered specificity that result from somatic hypermutation in vivo. PMID- 7499850 TI - Naturally processed peptides from rheumatoid arthritis associated and non associated HLA-DR alleles. AB - Naturally processed peptides from immunoaffinity-purified HLA-DRB1*0401, DRB1*0404 (rheumatoid arthritis (RA)-associated), and -DRB1*0402 (non-RA associated) molecules were analyzed by capillary liquid chromatography and mass spectrometry. The molecular weights observed for more than 60 eluted peptides from each HLA-DR protein ranged from 788 to 3535 atomic mass units, corresponding to peptides 7 to 32 amino acids in length. Sequencing of more than 60 of the abundant peptides revealed nested sets of peptides that were derived from only 12 different proteins. The majority of these proteins were membrane-associated (HLA class I, class II, and Ig molecules). Synthetic peptides, corresponding to endogenous peptide sequences, bound with high affinity (5 to 80 nM) to the HLA-DR molecules from which they were eluted. In addition, most were promiscuous binding peptides in that they also bound to other HLA-DR molecules. Truncations of eluted peptide sequences and alanine scanning mutational analysis of a Mycobacterium leprae peptide were used to identify the peptide residues involved in binding to DRB1*0404 and DRB1*0402 molecules. Furthermore, an invariant chain peptide was eluted from the DRB1*0402 molecules but not from the RA-associated molecules. The lack of invariant chain peptides from DRB1*0401 and DRB1*0404 molecules may contribute to the loading of autoantigen peptides into these molecules and to their association with disease. PMID- 7499851 TI - Mapping of the complement regulatory domains in the human factor H-like protein 1 and in factor H1. AB - The human factor H-like protein 1 (FHL-1) is composed of seven repetitive domains (short consensus repeats; SCRs) that are identical in sequence to the seven NH2 terminal SCRs of the complement regulatory protein factor H. We have identified the native FHL-1 protein as a 42-kDa human plasma protein by immunoblotting and by comparing the mobility to that of a recombinant FHL-1 protein. Here, we demonstrate the existence of two distinct co-migrating human plasma proteins that represent the 42-kDa FHL-1 protein and the previously identified 43-kDa factor H related 1 beta protein. Similar to factor H, the recombinant FHL-1 protein displays cofactor activity in factor I-mediated cleavage of C3b. To identify relevant SCRs of factor H and FHL-1, we recombinantly expressed the domains shared between the two proteins in the baculovirus expression system. Recombinant FHL-1 and all truncated forms that include SCRs 1 to 4 displayed cofactor activity. All four NH2-terminal SCRs are essential, as deletion mutants composed of SCR 1 and 4 only; of SCRs 1, 2, and 4 only, or of SCRs 1, 3, and 4 only were functionally inactive. Similarly, the distance between these individually folding domains is critical for function, as a recombinant protein that had two and four amino acids inserted between SCRs 1 and 2 or between SCRs 3 and 4, respectively, had no activity. These results demonstrate that all four NH2-terminal SCRs of FHL 1 (and of factor H) are required for cofactor activity in factor I-mediated cleavage of C3b, and that the distance between these SCRs is essential. PMID- 7499852 TI - Mouse surfactant protein-D. cDNA cloning, characterization, and gene localization to chromosome 14. AB - Surfactant protein-D (SP-D) is a collectin found associated with surfactant in the lung. SP-D has also been functionally characterized as an opsonin for diverse microorganisms and a chemoattractant for phagocytic cells. To determine the structure of mouse SP-D, we isolated and characterized clones from a B6/CBAF1J strain lung cDNA library using a PCR-derived genomic probe. The deduced sequence predicts a 19-amino acid signal sequence, a 25-amino acid long NH2 terminus with two cysteines, followed by an uninterrupted collagen domain with 59 Gly-X-Y repeats. Next, a short "neck" domain of 28 amino acids, with a potential to form trimeric alpha-helical coiled coil is found ending in a COOH-terminal 125-amino acid carbohydrate recognition domain. The mature mouse SP-D protein of 355 amino acids shows strong homology to rat (92% identity), human (76%), and bovine (72%) SP-D amino acid sequences. Northern blot and RT-PCR analysis revealed that the mouse SP-D gene is expressed predominantly in lung and, surprisingly, also in heart, stomach, and kidney but not in brain. In contrast, mouse surfactant protein-A (SP-A) mRNA expression was found to be restricted to lung. Human lung and stomach, but not heart or liver, were found to express SP-D mRNA, as determined by PCR. The mouse SP-D gene (Sftp4) has been localized to chromosome 14 (to a region syntenic to human chromosome 10), closely linked to the genes for other collagenous lectins, mannose-binding protein-A (MbI1), and SP-A (Sftp1). PMID- 7499853 TI - Targeted removal of the mu switch region from mouse hybridoma cells. A test of its role in gene expression in the endogenous IgH locus. AB - The switch regions adjoining the DNA encoding the Ig heavy chain constant regions have been implicated in gene expression as well as isotype switching, in that transgenic mice express switch-containing transgenes at a level 100- to 1000-fold higher than the corresponding switch-deleted transgenes. To test whether the switch region of the natural IgH locus is also required for high level expression we have used homologous recombination to generate targeted recombinant hybridoma cell lines that lack the switch region sequences from the major intron of the mu gene. The expression pattern of these switch knock-out cell lines was compared with that of the parental cell line as well as to that of control recombinants using both steady-state mRNA level and nuclear run-on activity to assess heavy chain gene expression. In striking contrast with the results reported for transgenic animals, we have found only a minimal effect, if any, of deleting the switch element from the natural chromosomal location. PMID- 7499854 TI - IL-12 promotes enhanced recovery from vesicular stomatitis virus infection of the central nervous system. AB - To investigate the role of a cytokine in host defense against the vesicular stomatitis virus (VSV) infection of the central nervous system (CNS), IL-12 was injected i.p. into groups of 10 BALB/c mice on days -1, 0, 1, 2, and 3 postinfection. Four days postinfection, mice were examined. IL-12 strongly enhanced immunity to VSV infection in the CNS as demonstrated by 1) decreased VSV titers in brain homogenate of IL-12-injected mice compared with those of controls; 2) increased expression of inducible nitric oxide synthase in the CNS; 3) enhanced expression of both MHC class I and class II Ags in the CNS; 4) increased T cell infiltration in the CNS, especially in the olfactory bulb; and 5) diminished VSV-induced apoptosis in olfactory bulb. No detrimental effect was observed even with the 200 ng/mouse dose of IL-12. Protective effects of IL-12 were dose dependent. Collectively, these results demonstrate that exogenously added IL-12, even when injected peripherally, significantly enhances recovery from VSV infection of the CNS. PMID- 7499855 TI - Endogenous IL-2 contributes to T cell expansion and IFN-gamma production during lymphocytic choriomeningitis virus infection. AB - IL-2-deficient mice were used to examine the role of endogenous IL-2 for supporting T cell proliferative responses during infection with lymphocytic choriomeningitis virus (LCMV). The studies showed that, although virus-specific CTL activity was induced in the absence of IL-2, the overall magnitude of the response was profoundly inhibited. Examination of proportions and numbers of CD8+ T cells demonstrated that the normal virus-induced expansion of these cells was virtually eliminated in spleens and dramatically decreased in lymph nodes from IL 2-negative mice. Absence of endogenous IL-2 also significantly inhibited virus induced activated T cell production of IFN-gamma, as well as increases in frequencies and numbers of IFN-gamma-producing cells. Reductions in immune responses were accompanied by impaired viral clearance. Although T cell responses were dramatically reduced in IL-2-deficient, as compared with IL-2-containing mice, activation signals were being delivered in vivo because induced CTLs were sensitive to the cell cycle-specific toxin, hydroxyurea (HU), and CD8+ T cells had induced expression of the IL-2R alpha- and beta-chains. These studies demonstrated that, although low levels of T cell responses can be induced in the absence of IL-2, the factor plays a unique and critical role in supporting T cell proliferative responses in vivo and in optimizing induction of the biologic functions mediated by these cells. Furthermore, the results identify a role for IL-2 in promoting IFN-gamma production in vivo. PMID- 7499856 TI - An ospA frame shift, identified from DNA in Lyme arthritis synovial fluid, results in an outer surface protein A that does not bind protective antibodies. AB - Passive immunization with murine or human Abs to outer surface protein A (OspA) can protect mice against Borrelia burgdorferi, but OspA Abs elicited during natural infection in mice or humans are unable to clear the spirochete from the infected host. To examine Ab binding by OspA during the course of human infection, we amplified the operon encoding full-length ospA and ospB from synovial fluids of a patient with chronic Lyme arthritis, the first such recoveries from human material, at four separate time points over 4.5 mo, and expressed OspA in Escherichia coli. OspA mAbs that passively protected mice from infection did not bind one of the expressed OspAs, because of a deletion in ospA that resulted in a frame shift and premature stop codon near the carboxyl terminus. However, expressed OspA from a later synovial fluid sample did not contain this deletion. Thus, although altered forms of OspA, which potentially can influence host immune effectiveness, do occur in the human host, they cannot be the only factors responsible for microbial persistence. PMID- 7499857 TI - Protective immunity induced by B7/CD28-costimulated gamma delta T cells to the EL 4 lymphoma in allogenic athymic mice. AB - We previously reported that the murine EL-4 lymphoma (H-2b) transduced with a retrovirus containing the murine B7-1 gene (B7+ EL-4) grew transiently for several weeks and subsequently regressed in allogenic BALB/c (nu/nu) athymic mice (H-2d). We now show that, in contrast, B7+ EL-4 cells grow progressively in several combined immunodeficiency mice, including SCID and NIH III mice, which lack T cells expressing either TCR-alpha beta or -gamma delta. Furthermore, depletion of gamma delta T cells with a specific mAb made possible the progressive growth of B7+ EL-4 cells in 90% of athymic mice while depletion of alpha beta T cells allowed tumor growth in 50% of these mice. Immunization of athymic mice with B7+ EL-4 cells prevented the outgrowth of wild-type B7- EL-4 cells. This protective immunity was abrogated by in vivo treatment with an anti TCR-gamma delta mAb, further indicating that gamma delta T cells play an important role in tumor rejection by athymic mice. A gamma delta T cell line, Tc1, was established from B7+ EL-4-immunized athymic mice by repeated restimulation in vitro with irradiated B7+ EL-4 cells. When tested against a broad spectrum of target cells, Tc1 lysed several murine lymphoma lines, but did not lyse other tumor lines, suggesting that the Ag recognized by Tc1 has a limited distribution. Our data demonstrate that gamma delta T cells, and, to a less extent, extrathymic alpha beta T cells, mediate an immune response against B7+ EL-4 cells in allogeneic athymic mice. PMID- 7499858 TI - Inhibition of vaccinia virus DNA replication by inducible expression of nitric oxide synthase. AB - Nitric oxide (NO) exerts multiple biologic roles in animal cells through differential regulation of three distinct forms of NO synthase encoded by separate genes. Macrophage-inducible nitric oxide synthase (iNOS) has been correlated with inhibition of viral growth, but little is known about the mechanism of this effect. To study the antiviral role of NO, we have generated a vaccinia virus (VV) recombinant expressing iNOS under the control of Escherichia coli LacI operator/repressor elements. When cultured cells of various origins are infected with this recombinant virus, there is inducible expression of iNOS in the presence of isopropylthio-beta-galactoside, as determined by Western blot and by detection of nitrite, a NO oxidation product. The levels of nitrite increase with time after infection, correlating with marked inhibition of VV DNA and late protein synthesis. Expression of VV early proteins is not affected by NO. Inhibition of VV DNA synthesis is likely to be in part a consequence of NO mediated inhibition of viral ribonucleotide reductase, as this inhibition can be partially overcome by addition of deoxyribonucleosides. Inhibition of the essential viral functions by NO results in a reduction of virus yields by 50 to 90%, depending on the cell line. Thus, our results demonstrate a direct antiviral effect of NO, with inhibition of VV replication occurring at the level of DNA synthesis. PMID- 7499859 TI - CD8 T cell clones inhibit antitumor T cell function by secreting IL-10. AB - We have reported that in irradiated, long-term surviving RFM strain of mice there is enhanced kinetics of tumor development upon challenge with RFM lymphoma cells. We reported that we cloned splenic oncofetal (OFA)-specific, noncytotoxic CD8+ T cells from such mice. These noncytotoxic CD8+ T cell clones secrete a factor upon Ag stimulation that inhibits the ability of OFA-specific RFM cytotoxic T (TC) cell clones from killing 5T RFM lymphoma cells in vitro. These supernatants do not inhibit the tumor cell-induced proliferation of the TC cell clones however. We report here that OFA-stimulated, RFM-noncytotoxic CD8 T cell clone culture supernatants also inhibit IFN-gamma-secretion by stimulated CD4 and CD8 RFM anti OFA effector T cell clones in a dose-dependent manner. The inhibitor in those culture supernatants acts in neither an Ag-specific nor MHC-restricted manner. We find that the culture supernatants of OFA-stimulated, noncytotoxic CD8 T cell clones contain IL-10, while those from OFA-stimulated, RFM OFA-specific TC cell clones do not. We show that monoclonal anti-IL-10 Ab specifically blocks the inhibition of cytotoxic activity and IFN-gamma secretion by OFA-specific CD8 and CD4 effector T cell clones in a dose-dependent manner in vitro. Incorporation of anti-IL-10 Ab into the cytotoxicity assays of the OFA-specific, noncytotoxic CD8+ T cell clones against 5T tumor cells restores their cytotoxic activity. This may suggest that one way of inducing anergic T cells is by induction of IL-10 secretion. PMID- 7499860 TI - Involvement of natural killer cells in nitric oxide production by spleen cells after stimulation with Mycobacterium bovis BCG. Study of the mechanism of the different abilities of viable and killed BCG. AB - Viable and killed BCG were compared for the ability to induce nitric oxide (NO) production in normal spleen cells and peritoneal exudate macrophages. Stimulation of spleen cells with viable BCG resulted in a strong expression of inducible NO synthase followed by an enhanced production of nitrite. However, those responses were not induced after stimulation with killed BCG or when macrophages were stimulated with killed and viable BCG. The same level of TNF-alpha mRNA expression was observed in reverse transcription-PCR after stimulation of spleen cells with viable and killed BCG. However, IFN-gamma production was induced only when spleen cells were stimulated with viable BCG. Concurrent stimulation of rIFN gamma with either viable or killed BCG resulted in a strong nitrite production by macrophages. Neutralization of IFN-gamma and TNF-alpha caused a complete inhibition of nitrite production. Furthermore, anti-asialo GM1 Ab plus complement treatment abolished IFN-gamma production after stimulation with viable BCG, indicating that the NK cell was the major source of IFN-gamma, and its production was triggered only by stimulation with viable BCG. The present study showed that the ability of viable BCG to induce NO production is superior to that of killed BCG, and both IFN-gamma and TNF-alpha are essential for BCG-induced NO production by spleen cells. NK cells appeared to be important as a source of IFN-gamma, and the insufficient NO induction by killed BCG was due to the inability to induce IFN-gamma from NK cells. PMID- 7499861 TI - Impact of interferon-gamma receptor deficiency on experimental Staphylococcus aureus septicemia and arthritis. AB - The role of IFN-gamma in the regulation of host resistance of Staphylococcus aureus was studied using IFN-gamma receptor-deficient (IFN-gamma R-/-) mice in a model of S. aureus-induced septicemia and arthritis. IFN-gamma R-/- mice and wild type controls were inoculated intravenously with a toxic shock syndrome toxin-1 producing S. aureus LS-1 strain. IFN-gamma R-/- mice displayed significantly more frequent and more severe arthritis compared with wild-type littermates (p < 0.01) throughout the course of infection. Notably, IFN-gamma R-/- mice developed severe sepsis with high mortality early after the inoculation with staphylococci. However, the mortality of wild-type mice became significantly higher at later stages of the disease compared with IFN-gamma R-/- mice (p < 0.05). This differential outcome of sepsis-related mortality was associated with deficiencies of bacterial elimination from blood and parenchymatous organs and correlated well to serum levels of IL-6 and spleen IL-1 beta and TNF-beta mRNA expression. Thus, bacterial growth and proinflammatory cytokines IL-1 beta, TNF-beta, and IL-6 were higher at the early stage of infection in IFN-gamma-/- mice but increased at the later stage in wild-type littermates. Our data indicate that the absence of IFN gamma R leads to harmful as well as beneficial effects in S. aureus infection, depending on the stage of the disease and the localization of the infection. PMID- 7499862 TI - IL-10 is involved in the protective effect of dibutyryl cyclic adenosine monophosphate on endotoxin-induced inflammatory liver injury. AB - The effects of exogenous cAMP, dibutyryl cAMP (DBcAMP) on LPS-induced liver injury were examined in mice made hypersensitive to LPS by treatment with i.v. injection of Propionibacterium acnes. In vivo administration of DBcAMP significantly protected P. acnes-treated mice from LPS-induced liver injury, including apoptosis of hepatocytes. DBcAMP significantly increased circulating IL 10 level in correlation with suppression of the TNF-alpha level after LPS challenge in P. acnes-treated mice. Treatment with anti-IL-10 mAb abrogated the protective effect of DBcAMP on LPS-induced liver injury. Similar to in vivo findings, addition to DBcAMP to in vitro culture of liver adherent cells from P. acnes-treated mice enhanced IL-10 synthesis after LPS stimulation. These results suggest that the increment in IL-10 production by liver adherent cells is involved in the protective effect of DBcAMP on LPS-induced inflammatory liver injury. PMID- 7499863 TI - Subcellular distribution of docking/fusion proteins in neutrophils, secretory cells with multiple exocytic compartments. AB - Neutrophils contain at least four distinct types of secretory organelles, which undergo exocytosis during infection and inflammation. The signaling pathways leading to secretion of individual granules and their kinetics of exocytosis vary greatly, causing temporal and regional differences in docking and fusion with the plasma membrane. As a step toward understanding the processes underlying differential granular secretion in neutrophils, we assessed the presence and distribution of a number of proteins reported to be involved in vesicular docking and/or fusion in other systems. Specific Abs were used for immunoblotting of cells fractionated by density gradients and free-flow electrophoresis, and for localization by confocal immunofluorescence and electron microscopy. Syntaxin 1, VAMP (vesicle-associated membrane protein)-1, synaptosome-associated protein-25 (SNAP-25), synaptophysin, and cellubrevin were not detectable in human neutrophils. In contrast, syntaxin 4, VAMP-2, and the 39-kDa isoform of secretory carrier membrane protein (SCAMP) were present. SCAMP was found mainly in secondary and tertiary granules and in a fraction containing secretory vesicles, but was virtually absent from the primary (lysosomal) granules. This profile is consistent with the proposed "post-Golgi" distribution of SCAMP. VAMP-2 was largely absent from primary and secondary granules, but concentrated in tertiary granules and secretory vesicles. This pattern of distribution parallels the increasing sensitivity of these exocytic compartments to intracellular free calcium. Accordingly, ionomycin induced translocation of VAMP-2 toward the plasma membrane. Syntaxin 4 was found almost exclusively in the plasma membrane, and it accumulated in lamellipodia of migrating cells. This regional accumulation may contribute to localized secretion into the phagosomal lumen. PMID- 7499864 TI - Clearance pathways of soluble immune complexes in the pig. Insights into the adaptive nature of antigen clearance in humans. AB - Efficient delivery of immune complexes (ICs) to the mononuclear phagocytic system, and subsequent IC processing, may prevent their potentially harmful effects in other tissues and may also be important in the development of humoral immune responses. In mice, rabbits, and primates, the liver and spleen are the main sites of IC clearance. It has been demonstrated previously that the pulmonary capillaries in the pig are lined with macrophages and that certain particulates, including bacteria, localize to this organ. In this study, we used gamma scintigraphy to explore the sites and kinetics of clearance of soluble IC comprising 123I-labeled hepatitis B surface Ag (HBsAg):porcine anti-HBsAg in the Large White pig. At t = 10 min after i.v. injection, 43 +/- 5% (mean +/- SE) IC localized in the lungs, and 36 +/- 6% counts in the liver. At t = 85 min, values were: lungs, 15 +/- 4% and liver, 29 +/- 2%. Findings were similar following intraarterial injection. Complement depletion resulted in more rapid initial IC clearance (t1/2 = 5 min), reduced lung uptake (23 +/- 3% at 10 min), and impaired IC catabolism. In normal animals, 5 to 7% injected IC bound to PBMCs, but no E binding was seen. A fall in PBMC numbers (46 to 59% of baseline), was observed following IC injection. These findings contrast with our previous observations using analogous IC in humans, in which we did not observe any change in peripheral blood leukocyte counts consequent upon complex processing, suggesting that in humans, Es may function as a buffering system for complement-bearing IC in the circulation, preventing their interaction with leukocytes bearing complement and FcR, and the potential activation of these cells. PMID- 7499865 TI - Controlled recruitment of monocytes and macrophages to specific organs through transgenic expression of monocyte chemoattractant protein-1. AB - Transgenic mice overexpressing the chemokine monocyte chemoattractant protein-1 (MCP-1) in the thymus and central nervous system have a higher number of mononuclear cells in those tissues than do control littermates. In the thymus, there is a modest increase in the number of Mac-1 and F4/80 positive cells, but no apparent change in the number of lymphoid cells. A more pronounced mononuclear infiltrate is detected in transgenic mice expressing MCP-1 in the brain. The vast majority of the recruited cells in the brain are monocytes and macrophages, as defined by light microscopy, and ultrastructural and immunohistochemical criteria. Such cells are found in a perivascular orientation with minimal parenchymal infiltration, possibly as a consequence of the accumulation of MCP-1 in the vessels, as shown by immunohistochemistry. The mononuclear cell infiltrate in the brain can be significantly amplified by LPS treatment, suggesting that the recruitment properties of MCP-1 can be potentiated by additional factors. PMID- 7499866 TI - Characterization of mutant forms of recombinant human properdin lacking single thrombospondin type I repeats. Identification of modules important for function. AB - Properdin is a serum glycoprotein that up-regulates the alternative pathway of complement by stabilizing the C3b-Bb complex. It also binds sulfated glycoconjugates, such as sulfatide, in vitro. Properdin is composed of cyclic dimers, trimers, and tetramers of a 53-kDa monomeric subunit. The monomer contains an N-terminal region of no known homology and six thrombospondin type 1 repeats (TSRs) of approximately 60 amino acids. To identify the regions of properdin important for function, we have expressed human properdin, and mutant forms each lacking a single TSR, in Chinese hamster ovary cells. In addition, limited tryptic digestion yielded "nicked" properdin by the cleavage of one peptide bond in TSR5. The structural and functional properties of these altered forms of properdin were investigated. Properdin "nicked" in TSR5 is unable to bind C3b but retains its overall structure and its ability to bind sulfatide. The removal of TSR5 prevents C3b and sulfatide binding. Properdin lacking TSR4 is unable to stabilize the C3b-Bb complex but is able to bind C3b and sulfatide, and shows the presence of monomers and dimers in an electron microscope. Properdin without TSR3 is able to stabilize the C3b-Bb complex, to bind C3b and sulfatide, and forms dimers, trimers, and tetramers. Properdin lacking TSR6 is unable to form oligomers. The N-linked carbohydrate of properdin is not required for oligomerization or stabilization of the C3b-Bb complex. The results implicate TSR5 in both C3b and sulfatide binding, and suggest that TSR4 may also be involved in stabilization of the C3b-Bb complex. PMID- 7499867 TI - Protein kinase C activation precedes arachidonic acid release during IgG-mediated phagocytosis. AB - Previous studies demonstrated that both protein kinase C (PKC) and arachidonic acid (AA) are required for IgG-mediated phagocytosis by human monocytes. We have characterized a calcium-independent "phagocytic" phospholipase A2 (designated pPL) that mediates arachidonic acid release. The present studies were designed to order PKC and pPL in the phagocytic signaling pathway. The PKC inhibitors staurosporine and calphostin C caused a coordinated decrease in phagocytosis of IgG-opsonized erythrocytes and arachidonic acid release. The PLA2 activators mastoparan and melittin restored phagocytosis to PKC-inhibited cells, but were ineffective in monocytes pretreated with the pPL inhibitor bromoenol lactone. Similarly, PKC activation with PMA and diacylglycerol enhanced phagocytosis in the absence, but not in the presence, of bromoenol lactone. These results indicate that pPL may be regulated by an upstream phosphorylation event. Thus, we examined the effects of Ab-opsonized glass bead ingestion, okadaic acid-mediated inhibition of phosphatases, and PMA treatment on the activity of pPL and on its distribution between the cytosolic and membrane-associated compartments. IgG opsonized erythrocytes and okadaic acid caused an overall increase in pPL activity, with a twofold increase in membrane-associated pPL. PMA treatment caused a 1.8-fold increase in membrane-associated pPL activity. Okadaic acid and PMA mimic IgG-opsonized erythrocytes with respect to membrane activation of pPL, suggesting that pPL activity may be regulated by PKC. Collectively, these results indicate that pPL activity is modulated by PKC during IgG-mediated phagocytosis, and that the PKC requirement can be bypassed by direct activation of pPL. PMID- 7499868 TI - Collagen-like complement component C1q is a membrane protein of human monocyte derived macrophages that mediates endocytosis. AB - The collagen-like C1q molecule, a subcomponent of the first component of complement, C1, is synthesized by macrophages (M phi). Previously, we have demonstrated that C1q is a membrane protein of guinea pig peritoneal macrophages (M phi). To extend this observation as a general biologic characteristic of M phi, we investigated human (hu) monocyte-derived M phi. Interestingly, surface labeling with the biotin derivative sulfosuccinimidyl-6-(biotinamido)-hexanoate of M phi, freshly isolated monocytes, lymphocytes, granulocytes, and myelomonocytic U937 cells revealed that C1q occurs only on the surface of M phi and not on the surface of the other cells types. Therefore, C1q appears to be a marker for differentiation into M phi. FITC-labeled, fixed Staphylococcus aureus coupled to membrane C1q via a monoclonal alpha-hu-C1q Ab were used to demonstrate that membrane C1q is capable of mediating phagocytosis. Various detergents (Nonidet P-40, digitonin, lubrol, and Triton X-114) were used to solubilize membrane C1q. Membrane C1q of hu M phi is tightly bound to or located in the intact membrane, since treatment of cells with acidic buffers ("acid strip") failed to remove C1q from the cell surface. However, repeated freezing and thawing of cells and washing of segregated membranes with buffer containing 1 M KCl and 3 M urea brought about a marked release of membrane C1q. PMID- 7499869 TI - Inducible binding of bioactive cathepsin G to the cell surface of neutrophils. A novel mechanism for mediating extracellular catalytic activity of cathepsin G. AB - Catalytically active cathepsin G that is bound to the cell surface of human neutrophils may play a variety of roles in normal neutrophil biology and in pathobiology associated with inflammation. In this study, we describe expression of neutrophil cell surface-bound cathepsin G in response to TNF-alpha and platelet-activating factor (PAF) under conditions in which minimal free release of cathepsin G is detected. TNF-alpha and PAF alone induced modest (two- to threefold) increases in cell surface-bound cathepsin G, but exhibited a marked dose- and time-dependent priming effect for subsequent chemoattractant-induced responses (up to 15- to 25-fold increases in cell surface expression). When optimally primed (TNF-alpha, 100 U/ml, or PAF, 10(-9) M), neutrophils expressed five- to sixfold more cell surface-bound cathepsin G, in comparison with cells exposed to FMLP alone. Priming responses were more rapid with PAF (15 s to 5 min) than with TNF-alpha (1 to 60 min). Optimally primed and FMLP-stimulated neutrophils express approximately 160 ng of catalytically active cathepsin G per 10(6) cells, which represents approximately 11% of the cellular content of unstimulated cells. Cathepsin G binds to the cell surface by a charge-dependent mechanism since: 1) incubation of cells with highly positively charged molecules abrogated agonist-induced up-regulation of the cell surface expression of cathepsin G and 2) cathepsin G was eluted from the cell surface by high concentrations of NaCl. These data indicate that interactions between biologically relevant pro-inflammatory cytokines and chemoattractants serve to markedly up-regulate cell surface-bound cathepsin G. The focused catalytic activity of cell surface-bound cathepsin G may alter endothelial and epithelial barriers, promote thrombogenesis, injure extracellular matrix, and/or facilitate directed migration of neutrophils during inflammation. PMID- 7499870 TI - Identification and characterization of a novel surface antigen gene induced in mast cells activated through the high affinity IgE receptor. AB - In an effort to isolate novel genes involved in inflammation and/or mast cell activation, we have used a combination of differential screening and subtractive hybridization to isolate genes whose expression are induced upon activation of a transformed rat mast cell line. One of the isolated clones, pMCA-32, contained an open reading frame of 278 amino acids that included a putative hydrophobic transmembrane domain, a cysteine rich Ig-like extracellular domain, and a cytoplasmic domain containing three consensus SH2-domain phosphotyrosine binding sites. The MCA-32 gene is also highly conserved between rat and mouse, with the two coding regions being 73% identical. Although the MCA-32 coding region did not contain an obvious signal peptide, MCA-32 protein was detected on the surface of rat mast cells, and the cloned cDNA produced a cell surface protein when expressed in COS-7 cells. MCA-32 RNA from both mouse and rat undergoes alternative splicing, producing an mRNA containing an in-frame deletion of the TM domain, suggesting that a form of MCA-32 protein may be secreted. MCA-32 mRNA expression was up-regulated upon activation of RBL-2H3 cells and was highly abundant in primary peritoneal mast cells. Expression of MCA-32 RNA was only observed in primary and transformed mast cells from rat, while in the mouse MCA 32, RNA was also produced in significant amounts by a number of transformed monocyte cell lines. Thus, MCA-32 is a novel surface protein whose structure and expression suggest roles in the development and/or activation of mast cells and monocytes. PMID- 7499871 TI - Autocrine/paracrine involvement of platelet-activating factor and transforming growth factor-beta in the induction of phosphatidylserine recognition by murine macrophages. AB - The specific recognition of phosphatidylserine (PS) by macrophages is believed to be one means by which effete and apoptotic cells expressing PS on their outer membrane leaflet are targeted for phagocytosis. The aim of this study was to better understand the autocrine/paracrine factors involved in beta-glucan induction of PS recognition by macrophages. We provide evidence that both platelet-activating factor (PAF) and TGF-beta are involved in beta-glucan induction of PS recognition. This is based on the observations that the PAF receptor antagonist WEB 2086 and Ab against TGF-beta each could partially inhibit beta-glucan-induced PS recognition when used alone and could completely inhibit induction when used in combination. PAF, like TGF-beta, was found to prime macrophages for PS recognition, which could then be triggered by costimulation with a nonspecific phagocytic stimulus, latex particles. We also provide evidence that the priming by PAF and that by TGF-beta can occur independently of each other. This is based on the observations that 1) PAF priming was not blocked by anti-TGF-beta Ab, nor was TGF-beta priming prevented by WEB 2086; and 2) PAF did not increase the steady state level of TGF-beta mRNA, and TGF-beta did not induce PAF synthesis in these cells. PMID- 7499872 TI - Augmentation or inhibition of IFN-gamma-induced MHC class II expression by lipopolysaccharides. The roles of TNF-alpha and nitric oxide, and the importance of the sequence of signaling. AB - MHC class II expression on macrophages is one determinant of Ag presentation and the vigor of CD4+ T cell immunity. We show that LPS may either inhibit or augment IFN-gamma-induced MHC class II on macrophages depending on the sequence of the IFN-gamma and LPS signals. LPS inhibited MHC class II when added simultaneously with IFN-gamma, but augmented class II expression when added after IFN-gamma. Inhibition was due to nitric oxide (NO), which was only produced if LPS was given simultaneously with IFN-gamma. However, even when NO production was inhibited, LPS given simultaneously with IFN-gamma did not augment MHC class II expression. This suggests that LPS delivers different signals when given simultaneously vs after IFN-gamma. LPS augmentation of class II expression was functionally important because it correlated with increased Ag presentation. Augmentation by LPS of IFN-gamma-induced class II expression by macrophages has not been previously reported. We found that TNF-alpha, like LPS, inhibited IFN-gamma induced class II expression if NO was produced, but augmented it in the absence of NO formation. Studies with a neutralizing anti-TNF-alpha Ab, however, indicate that LPS augmentation of MHC class II did not require TNF-alpha. LPS augmentation involved a different mechanism than IFN-gamma induction of MHC class II. LPS augmentation occurred at a post-transcriptional level, whereas IFN-gamma induction occurred at the level of gene transcription. LPS augmentation was apparent after 2 h of stimulation by LPS, while IFN-gamma induction of class II expression required more than 8 h. PMID- 7499873 TI - Oligoclonality of lung T lymphocytes following exposure to allergen in asthma. AB - We were interested in studying the lung allergen-specific T cell repertoire in different conditions of allergen exposure in subjects with atopic asthma. Twenty one allergic individuals were studied: 17 subjects suffering mainly from asthma and 4 from rhinitis. They all performed spirometry and methacholine challenge. All patients were subjected to bronchoalveolar lavage (BAL), either at base line (no challenge) or after allergen or histamine challenge, and the TCR repertoire of their lung T cells was studied with heteroduplex analysis. Expansion of single T cell clones was observed in one (of seven) asthmatic subject that reported a recent exposure to allergen and had high bronchial hypersensitivity to methacholine, and in seven/seven asthmatic subjects who underwent BAL after they suffered an early asthmatic reaction to experimental allergen inhalation. Remarkably, ex vivo expanded clones included allergen-specific T cells. In two of the seven subjects who underwent BAL after allergen challenge, two different lung segments were lavaged. A strikingly symmetrical distribution of the expanded clones was found in these samples. Control subjects and six of seven asthmatic patients studied at base line showed polyclonality of lung T cells. In conclusion, T lymphocytes are clonally expanded in the lower respiratory tract only in asthmatic subjects exposed to allergen. These results suggest that in allergic asthma, the inhalation of sensitizing allergens can recruit to the lung T lymphocytes that include allergen-specific T cell clones. PMID- 7499874 TI - Primary hormonogenic sites as conserved autoepitopes on thyroglobulin in murine autoimmune thyroiditis. Secondary role of iodination. AB - We hypothesized earlier that conserved T cell epitopes and those unique to mouse thyroglobulin (MTg) contributed to its total thyroiditogenicity in murine autoimmune thyroiditis. Recent studies of synthetic peptides from human Tg (HTg) revealed no immunodominant epitopes. The role of iodine residues, considered by some to render Tg immunogenic, became unclear, since only one 12-mer peptide contained thyroxine (T4) positioned at hormonogenic site 2553. It primed T cells for thyroiditis transfer, but noniodinated peptide containing thyronine (T0) was not compared. To determine 1) whether other primary hormonogenic sites were likewise immunogenic and 2) whether iodination was requisite for this and other sites to be an autoepitope, we derivatized three pairs of 12-mer peptides, 1-12, 2549-2560, 2559-2570, containing T0 or T4 at positions 5, 2553, or 2567, respectively. The six peptides were used to stimulate MTg-primed cells in vitro and to immunize mice. None directly induced thyroiditis; peptide Abs were the lowest in mice given hT0(2567) or hT4(2567). Of the three T4-containing peptides, hT4(5) and hT4(2553), but not hT4(2567), stimulated MTg-primed or HTg-primed T cells in vitro, with hT4(2553) being the stronger. Comparing hT0(2553) with hT4(2553), both activated MTg-primed, or peptide-primed, T cells to transfer thyroiditis. The marked immunogenicity of noniodinated hT0(2553) and the poor antigenicity of hT4(5) and hT4(2567) demonstrate that immunogenicity of a conserved hormonogenic site is dependent more on its amino acid sequence than on T4 substitution. Iodination may enhance antigenicity and/or binding affinity, but it is not required for a Tg hormonogenic site to be an autoepitope. PMID- 7499875 TI - Antiviral cytotoxic T lymphocytes in vaginal mucosa of simian immunodeficiency virus-infected rhesus macaques. AB - The mucosal immune system of the female reproductive tract is of central importance for protection against sexually transmitted diseases, including HIV; however, this arm of the immune system remains poorly understood. Antiviral CTL responses never have been documented in the genital tract and the role of CTL in this anatomic site is unknown. In this study, CD8+ intraepithelial lymphocytes (IEL) in the vaginas of six simian immunodeficiency virus (SIV)-infected female rhesus macaques were identified by immunohistochemistry to be CD2+ and TCR beta chain+. In addition, the majority of CD8+ IEL contained TIA-1+ cytoplasmic granules that are associated with CTL activity. CD8+ T cells were isolated from the vaginal epithelium and submucosa and amplified by limiting dilution in the presence of feeder cells. SIV p55gag and/or gp160env-specific lysis was detected in cultures of vaginal epithelial but not submucosal CD8+ T lymphocytes. Estimated SIV-specific precursor CTL frequencies were higher in the vaginal CD8+ IEL population of chronically infected monkeys than in the same cells from acutely infected monkeys or a naive control monkey. These results provide the first demonstration that antiviral CTL are present in the vaginal epithelium, and suggest that a vaccine may be able to generate anti-HIV CTL in the genital mucosa. PMID- 7499878 TI - Quantitative cell line based bioassays for human cytokines. AB - Research in cytokine biology is ever increasing and it is clear that cytokines are involved in a wide range of pathological and physiological processes. The validity of such research relies heavily on the appropriate measurement of levels of cytokines in various biological samples. Amongst the currently available methods for measuring cytokine levels, it is only the biological assay of samples that can directly provide estimates of biologically active cytokines present in test samples. Of the several bioassay systems available for detecting cytokines, cell line based bioassays are the easiest to perform and provide the most precise and accurate data. The suitability of any cell line for bioassaying a particular cytokine depends on several criteria such as sensitivity, ease of growth maintenance, and cytokine specificity. The design and analysis of cell line bioassays is also important in providing valid estimates of cytokine levels. We review the most useful cell lines currently available for bioassaying cytokines and discuss the design advantages and limitations of cytokine bioassays. PMID- 7499877 TI - In vivo clonotypic regulation of human myelin basic protein-reactive T cells by T cell vaccination. AB - Autoreactive T cells specific for myelin basic protein (MBP) are implicated in the pathogenesis of multiple sclerosis. The mechanism by which MBP-reactive T cells are regulated in vivo remains unknown, but is thought to involve the clonotypic regulatory network that can be induced by immunization with attenuated T cells and TCR peptides. We reported previously that immunization of multiple sclerosis patients with irradiated MBP-reactive T cells (T cell vaccination) induced T cell responses to the immunizing clones, resulting in a clonal depletion of circulating MBP-reactive T cells. In this study, we demonstrated that in the majority of the recipients, MBP-reactive T cells remained undetectable in circulation over a period of 1 to 3 yr after vaccination, while they reappeared in some individuals (three of nine), coinciding with clinical exacerbation. The reappearing MBP-reactive T cells were found to originate from clonal origins different from those of T cells persisting before immunization, suggesting a shift of the T cell repertoire to other determinants of MBP. The immunization induces predominantly CD8+ regulatory T cells capable of lysing the immunizing clones in a clonotype-specific manner. The T cell responses induced by immunization were restricted to the immunizing clones and did not affect MBP reactive clones not used for immunization. Our data further suggest that different hypervariable regions of the TCR may be involved in the observed clonotypic interaction. This study provides useful information for designing future clinical trials using T cell vaccination and other TCR-based therapeutic strategies. PMID- 7499879 TI - Mathematical considerations of competitive polymerase chain reaction. AB - Reverse transcriptase polymerase chain reaction (PCR) is used frequently to monitor gene expression. It is generally regarded as a qualitative technique, although refinements have been made to improve quantification. The object of this study was to develop competitive PCRs to allow reliable quantification of the rat T cell cytokines interferon-gamma (IFN-gamma), interleukin-2 (IL-2) and interleukin-4 (IL-4). Truncated constructs of cDNA for these cytokines were prepared using appropriate pairs of standard and specially constructed primers designed to allow subsequent co-amplification of the purified competitor construct and the target cDNA. A high resolution capillary electrophoresis (CE) system was used for PCR product detection. The performance of the system was compared with a mathematical model that describes and predicts the exponential nature of the PCR reaction. Co-amplification of the competitor and target were achieved. A high level of resolution and accuracy was achieved using CE to detect and quantify the PCR products. The rates of generation of the respective products conformed closely but not exactly to the predictions of the mathematical model. The competitive PCRs estimated initial numbers of target cDNA within 1.1-5.0-fold relative to the amount of starting material as assessed by conventional spectrophotometric absorbance prior to dilution and amplification. A convenient and flexible competitive PCR strategy has been developed with accurate resolution of products and reliable quantification. Assay variability was far less than biological variability likely to be encountered in experiments investigating immunological responses in rats or other animals. PMID- 7499876 TI - Myelin antigen-coupled splenocytes suppress experimental autoimmune encephalomyelitis in Lewis rats through a partially reversible anergy mechanism. AB - Mechanisms of adult tolerance induced by injecting myelin Ag/ECDI (ethyl carbodiimide)-coupled splenocytes (Ag-SPL) were evaluated in Lewis rat experimental autoimmune encephalomyelitis (EAE). Rats could be tolerized against the major encephalitogenic epitope of guinea pig basic protein (Gp-BP), residues 72-89, using either S72-89-SPL or crude spinal cord homogenate (SCH)-SPL. In contrast to lymph node responses that were not affected significantly, the proliferation responses of blood T cells were markedly inhibited at the peak of EAE and during the recovery period to both Gp-BP and S72-89, but not to purified protein derivative (PPD), demonstrating Ag-specific tolerance. Tolerance induction reduced the number of infiltrating spinal cord (SC) cells, especially recruited CD45RC+ cells, as well as SC proliferation responses to S72-89 throughout the course of EAE. In contrast, SC response to PPD was increased at onset of EAE, but later during recovery the PPD response was also decreased compared with control rats. Tolerance induced by S72-89-SPL in blood and SC T cells could be reversed by incubation in IL-2, in accordance with an anergy model. BP-specific T cells preincubated in vitro with Gp-BP-SPL were rendered unresponsive to Gp-BP or S72-89, compared with the same T cells preincubated with histone (Hist)-SPL that remained Ag responsive. Consistent with an anergy model, preincubation with BP-SPL+IL-2 partially prevented tolerance induction to BP. T cells tolerized in vitro to BP-SPL induced milder EAE with delayed onset compared with control-tolerized T cells that produced lethal disease. These results demonstrate the efficacy of myelin Ag-coupled SPL in preventing EAE by selective tolerization of encephalitogenic T cells through a partially reversible anergy induction mechanism. PMID- 7499880 TI - A new adhesion assay using buoyancy to remove non-adherent cells. AB - A new adhesion assay was developed that utilizes buoyancy, rather than washing or centrifugation, to remove non-adherent cells. Biotinylated cells were added to wells containing cell monolayers or purified protein substrates. Non-adherent cells were then removed by floatation on a dense Percoll solution. The adherent cells were fixed tightly to the plate with a Percoll/glutaraldehyde fixative and quantitated by streptavidin: horseradish peroxidase chemistry. In a side-by-side comparison of buoyancy and washing assays, the buoyancy method detected B16F10 binding to purified fibronectin at a 4-fold lower fibronectin concentration and human umbilical vein endothelia cell (HUVEC) binding to laminin at a 10-fold lower laminin concentration than did washing assays. In cell to cell adhesion assays, the buoyancy method was able to detect significantly greater binding of mononuclear leukocytes and KM12-L4 colon carcinoma cells to IL-1 beta treated human umbilical vein endothelial cells (HUVEC). The binding of human promyelocytic leukemia HL60 cells to control and IL-1 beta treated HUVEC was the same (approximately 60%) with the buoyancy method, while a washing assay demonstrated 8-fold higher binding (51% vs. 6%) of HL60 on IL-1 beta treated cells. The buoyancy assay is useful for detecting weak cell to protein adhesion and may be useful for detecting cell to cell adhesion when background binding is sufficiently low. PMID- 7499882 TI - Immunoblot analysis of bacterial polysaccharides: application to the type specific polysaccharides of Streptococcus suis and Streptococcus agalactiae. AB - A method for the immunoblot analysis of the type-specific capsular polysaccharides of streptococci was developed. The capsular polysaccharides were extracted by sonication and subjected to polyacrylamide gel electrophoresis in the absence of sodium dodecyl sulphate (SDS). After electrophoresis the polysaccharides were transferred to charged PVDF-N membranes and probed with the type specific antibodies. A characteristic ladder-like pattern of polysaccharide bands specific for the serotype (1, 2, 4, 7) was observed for capsular extracts of Streptococcus suis. Human immune sera against type-specific group B streptococcal polysaccharides reacted most strongly with the immunizing polysaccharide type (Ia, II, III). The previously observed crossreactions between the group B streptococcal type-specific capsular preparations were shown to be due to binding to the isomeric polysaccharide molecules. Thus, gel electrophoresis combined with immunoblot analysis seems to provide a novel method for the molecular and immunochemical characterization of bacterial polysaccharides and for the study of the specificity and properties of antibodies to capsular polysaccharides. PMID- 7499881 TI - Immunoglobulin A subclass measurement in serum and saliva: sensitivity of detection of dimeric IgA2 in ELISA depends on the antibody used. AB - The existence of two IgA subclasses in humans has been reliably shown by biochemical, immunochemical and genetic means. IgA is unique among immunoglobulins in the regular occurrence of both monomeric and polymeric forms. In order to be able to study the relationship between monomeric and polymeric IgA1 and IgA2 concentrations in the circulation and mucosal compartment i.e. secretions, it is essential that the methods used are not biased by the molecular size of the IgA under investigation. We validated IgA and IgA subclass measurements in serum and saliva by sandwich enzyme-linked immunosorbent assay (ELISA). Coating reagents were specific mAbs against IgA (clone 4E8), IgA1 (clone 69-11.4) or IgA2 (clone 16-512-H5 and clone IF8.58). Pooled normal human serum and purified dimeric IgA1 (d-IgA1) or IgA2 (d-IgA2) myeloma proteins were used to standardize the assays. Polymeric and monomeric forms of IgA in sera from volunteers and patients with myelomatosis were assayed in fractions separated by high performance liquid chromatography (HPLC). Dithioerythritol (DTE) was used to determine the influence of the quarternary structure of IgA on its detection by mAbs. We found that mAbs 4E8, 69-11.4 and 16-512-H5 reliably measured d-IgA, d IgA1 and d-IgA2 respectively, independent of the standard employed. Clone IF8.58 underestimated the concentration of d-IgA2 (correction factor +/- 2) with increased sensitivity in the presence of DTE. This difference is probably explained by the composition of the immunogen against which the mAb was raised. We conclude that no reliable conclusions can be made concerning the subclass ratio in biological fluids unless the monoclonal antibodies used have been appropriately validated. PMID- 7499883 TI - Scanning electron microscopy of immunogold labeled cat allergens (Fel d 1) on the surface of airborne house dust particles. AB - This study investigated the ability of an immunogold labeling technique to demonstrate the presence of Fel d 1 (domestic cat) allergens on the surface of particles in samples of airborne house dust. Suspended particulate matter was sampled from ten Norwegian households, five with and five without a domestic cat. The specimens were immunogold labeled and examined in the backscatter electron imaging mode of the scanning electron microscope and in the transmission electron microscope. X-ray microanalysis was also applied to execute element analysis of the suspended particular matter. The gold probe was mainly detected on carbon particles in the suspended particulate matter, both on small (< 1 microns) and larger carbon aggregates (1-10 microns). The present method may be useful in studying the localisation of different allergens on airborne house dust particles of various sizes and composition. PMID- 7499884 TI - Quantification of measles virus by a virus receptor-dependent and haemagglutinin specific T cell stimulation assay. AB - The human measles virus receptor CD46 plays a major role in the uptake of measles virus (MV) for antigen presentation by major histocompatibility complex class II molecules to T cells. On this basis, a new bioassay has been set up to quantify measles virus in a cell free tissue culture supernatant. A stable mouse B cell transfectant expressing CD46 was used as the antigen presenting cell for presentation of measles virus to a haemagglutinin-specific and class II restricted mouse T cell hybridoma. The measles virus haemagglutinin was quantified by its ability to stimulate IL-2 secretion by the T cells. A good correlation was found between the amount of haemagglutinin measured in supernatants from infected cells using the CD46-dependent T cell stimulation assay and the number of infectious viral particles as determined in a plaque assay. When MV was purified on a discontinuous sucrose gradient, most of the infectious virus and the haemagglutinin antigen were recovered in the same fraction. These data indicate that the CD46-dependent haemagglutinin-specific T cell assay could be used to measure the production of measles virus in the supernatant of infected cells. The assay required only 48 h, was sensitive, highly specific, and did not rely on the replication of the virus. This new bioassay would be applicable for the detection of any other virus provided that antigen presenting cells expressing the corresponding virus receptor and virus envelope glycoprotein-specific T cells are available. Moreover, it would be an interesting tool to monitor the receptor binding properties of attenuated vaccine virus and envelope glycoprotein subunit vaccines. PMID- 7499885 TI - Detection of anti-liver cytosol antibody type 1 (anti-LC1) by immunodiffusion, counterimmunoelectrophoresis and immunoblotting: comparison of different techniques. AB - Liver cytosol specific antibody type 1 (anti-LC1) was first described in a proportion of patients with liver/kidney microsomal antibody type 1 (anti-LKM1) positive autoimmune hepatitis (AIH) and is routinely evaluated by immunodiffusion (ID). Using human liver cytosol as the source of antigen, we have used ID, counterimmunoelectrophoresis (CIE) and immunoblotting (IB), to test sera from 167 patients with documented chronic liver diseases of different etiology. 15 patients had antinuclear antibody (ANA) and/or smooth muscle antibody (SMA) positive AIH, 13 had anti-LKM1-positive AIH, four had ANA/SMA/anti-LKM1-negative AIH, 76 had anti-LKM1-positive hepatitis C (recently renamed unclassified chronic hepatitis-UCH), 40 had chronic hepatitis C, 15 had chronic hepatitis B, and 4 had chronic hepatitis D. A precipitin line of identity with an anti-LC1 reference serum was detected both by ID and CIE in 16 patients: six with anti-LKM1-positive 'definite' AIH, four with ANA/SMA/anti-LKM1-negative 'definite' AIH, and six with anti-LKM1-positive UCH. By IB, 14 out of the 16 anti-LC1-positive sera (87.5%) reacted with a 58 kDa human liver cytosolic polypeptide, whereas three out of 16 (19%) recognised an additional 60 kDa band. Compared to ID, CIE is more economical in terms of both time and reagents and provides more clear-cut results. The 58 kDa reactivity by IB was detectable in nearly all CIE/ID anti-LC1 positive patients, was not found among CIE/ID anti-LC1-negative patients. In conclusion, CIE is the ideal screening test for the detection of anti-LC1, an autoantibody that can be regarded as an additional serological marker of AIH and is especially useful in ANA/SMA/anti-LKM1 negative cases. PMID- 7499887 TI - Simultaneous quantitation of multiple cytokine mRNAs by RT-PCR utilizing plate based EIA methodology. AB - Cytokines are small protein hormones produced during an immune response that are responsible for mediation and regulation of many aspects of immunity. Measurement of cytokines by several different methods has led to a broader understanding of the immune response. This paper describes a sensitive, reproducible, and quantitative RT-PCR assay for the simultaneous measurement of multiple cytokines. The main features of the methodology are: RNA competitors which control for all aspects of the process from RNA extraction, through reverse transcription (RT) and PCR amplification; a general cloning vector, pQPCR1, for building RNA competitors that does not require prior analyte cDNA cloning; and analysis by plate based EIA. This RT-PCR-EIA system is shown to be more sensitive than agarose gel electrophoresis followed by EtBr staining, measuring PCR product in the sub-nanogram range. It also extends the linear dynamic range of detection to a four log fold range of analyte concentration. The assay is reproducible, with coefficients of variation (CVs) in the 10-20% range. Moreover, the cloning vector is designed to accommodate multiple primer templates, thus allowing simultaneous quantitation of many different analytes from a single RT reaction. The described system is versatile and adapts to numerous analytes. PMID- 7499886 TI - Comparison of four lymphocyte isolation methods applied to rodent T cell subpopulations and B cells. AB - The aim of this study was to establish the validity of four lymphocyte isolation methods. The effects of three different erythrocyte lysing methods commonly used in the analysis of human cells, namely, lysis by ammonium chloride (AC), Becton Dickinson lysis (BDL) and the Coulter Q-Prep (CQP) preparation system were established by flow cytometry on rat lymphocyte subsets. The results were compared with those obtained with a Ficoll-Isopaque (FI) density gradient procedure adapted for use with rat cells. Lymphocyte isolation by AC or FI gradient was performed before labelling the lymphocyte subpopulations, whereas the BDL and CQP methods were performed after staining the cells in whole blood. The FI gradient yielded the lowest CD5+, CD4+ and CD25+ cell percentages. On the other hand AC lysis produced higher percentages of T cells and lower percentages of B cells than the other methods studied. The percentages obtained after BDL or CQP methods for T lymphocyte subsets and B cells were found to be reproducible. The commercial methods (BDL and CQP) are faster but rather expensive, whereas AC lysis and FI gradient separations are cheap and particularly useful when there is a requirement to culture the cells. PMID- 7499888 TI - Clinical correlates and substrate specificities of antibodies exhibiting neutrophil nuclear reactivity--a methodological study. AB - In a serological laboratory with a routine service for determining autoantibodies to human neutrophils, antibodies giving a selective or preferential reaction with the nucleus or perinuclear area of neutrophils are not uncommon. The aim of this study was to look for clinical correlates with the presence of such neutrophil reactive autoantibodies. The specificity of such antibodies for nuclear or cytoplasmic antigens was studied in 65 consecutive sera displaying nuclear/perinuclear reactivity at a titre of at least 80 using the indirect immunofluorescence technique (IIF) on ethanol-fixed leucocytes. The sera were also investigated by IIF on formalin-acetone fixed leucocytes and on HEp-2 cells. ELISA techniques were used to measure antibodies to azurophil granule constituents (ANCA), purified myeloperoxidase (MPO-ANCA), and lactoferrin (LF ANCA). Furthermore a qualitative spot immunoassay was used for the detection of antibodies to alpha, beta, and gamma fractions, and the nuclear fraction of neutrophils, purified proteinase 3 (PR3), MPO, enolase, lysozyme, elastase, lactoferrin, and cathepsin G. The diagnoses linked to such GS-ANA/pANCA positivity were arthritides, vasculitides, inflammatory bowel disease and chronic hepatic conditions. MPO was the main antigen recognized in the vasculitis group, but apart from that, rather limited antigen reactivity was demonstrable by these techniques, lysozyme being the most frequently recognized autoantigen in patients with arthritides. Human lymphocytes served as a suitable control substrate when distinguishing between GS-ANA/pANCA and ANA, whereas HEp-2 cells usually could not be used if both classes of antibodies were present in a sample. Furthermore, formalin-acetone fixation is not recommended for routine use. PMID- 7499889 TI - The assessment of antibody affinity distribution by thiocyanate elution: a simple dose-response approach. AB - We describe a simple dose-response approach to assess the affinity distribution of polyclonal antibodies. The proportion of antigen-specific antibodies dissociated by increasing concentrations of the mild chaotropic agent ammonium thiocyanate (NH4SCN) was measured by enzyme immunoassay, and the distribution of tolerances to this agent was presented in a histogram form. Such 'tolerance distribution', which is analogous to that described in classical dose-response bioassays, is proposed as a representation of the actual antibody affinity distribution. To test this approach, we assessed affinity maturation patterns of anti-Plasmodium falciparum IgG antibodies in paired sera obtained from 22 malaria patients during the acute infection and convalescence. We obtained patterns of antibody affinity distributions consistent with those previously described in immunization experiments with the aid of more complex laboratory and computational approaches. Therefore, we suggest the thiocyanate elution technique as an alternative method for rapid assessment of affinity distributions of polyclonal antibodies elicited against complex antigens, readily applicable to large number of serum samples. PMID- 7499890 TI - Oral rehydration solution. PMID- 7499891 TI - ORT programme and IMA. 1988. PMID- 7499893 TI - Management of persistent diarrhoea. PMID- 7499892 TI - Oral rehydration therapy programme in India: standard case management of acute watery diarrhoea. PMID- 7499894 TI - Feeding advice for children with acute diarrhoea. PMID- 7499895 TI - Role and responsibility of paediatricians in diarrhoea case management. PMID- 7499896 TI - Management of dysentery. PMID- 7499897 TI - Cholera outbreaks: role of oral rehydration therapy. PMID- 7499898 TI - Patterns of use of oral rehydration therapy in an urban slum community. PMID- 7499899 TI - Experiences in case management of diarrhoea in large hospitals. PMID- 7499900 TI - Role of physicians in preventing diarrhoea. PMID- 7499901 TI - Prevent diarrhoea deaths. Advice to the mother. PMID- 7499902 TI - What to do with OCA results from multiple angiographic views? PMID- 7499904 TI - Objective methods for optimizing JPEG compression of coronary angiographic images. AB - Digital angiographic images contain a significant amount of redundancy as well as some irrelevant information and noise. Therefore, it is possible to reduce the number of bits required to represent an image considerably. The lossy JPEG standard may be used provided that no significant diagnostic information is lost. As implemented in presently available hard- and software in most cases the luminance quantization table (LQT) is applied for gray level images, which may only be scaled by a so-called quality factor. The questions arise whether it is possible and worthwhile to specify quantization tables for the particular characteristics of angiograms. To assess the quality performance quantitatively, global numerical quality measures and evaluations based on Hosaka-plots were performed. Those diagrams compare the errors introduced into areas of different local activity. By the newly introduced weighting of these errors with the relative occupancy of the respective classes of activity the results got more reproducible. The blocking and blurring effects introduced by lossy JPEG compression could be compared objectively. Two new quantization tables were derived from the transfer function of the angiographic X-ray system, the modulation transfer quantization table (MTQT) and the star pattern quantization table (SPQT). Both tables guarantee that the blurring of sharp edges is minimized so that no deterioration around a coronary lesion occurs. Based on the signal-to noise ratio, the overall quality performance is the same as for the LQT. A general relation between the bit rate of the compressed image and the quality factor has been determined for images of high local activity and normally scaled coronary angiographic images (512 x 512). PMID- 7499903 TI - Does it matter how coronary projections are combined to assess restenosis following PTCA? AB - Three methods were used to combine measurements from biplane coronary views to assess restenosis following PTCA. The first 153 patients from the EMPAR trial with 118 measurable coronary stenoses were chosen for this study. The mean difference between the RAO and LAO projections used was 88 +/- 22 degrees (S.D.). There was no systematic difference in coronary dimensions between RAO and LAO views for the group. Coronary restenosis was primarily defined as loss of 50% or more of the luminal diameter gain from the procedure. The ratio of luminal loss to gain was calculated for each stenosis using (Method 1) only the lesser diameter of the two views, (Method 2) an average of the two views and, (Method 3) a calculated cross-sectional coronary lumen area. Rates of restenosis calculated in this way ranged between 36 and 44% amongst the three methods, which was not significantly different. However, individual coronary lesions might be classified as either restenosed or not by this formula, depending on which method was used to integrate the biplane coronary measurements. PMID- 7499905 TI - Digital high frame rate stress echocardiography for detection of coronary artery stenosis by high dose dipyridamole stress testing. AB - Diagnostic accuracy of high dose dipyridamole stress echocardiography (0.84 mg i.v./kg) for detecting coronary artery stenosis was assessed in 94 patients undergoing coronary angiography, and adverse effects were registered in the total study population of 120 patients. Echocardiographic analysis was performed with digital systolic cineloops with high frame-rate (47 frames/sec) for optimal left ventricular wall motion display. Results showed sensitivity of 73% for detection of arterial luminal stenosis > or = 75% or retrograde collateral flow to an occluded coronary artery. Sensitivity for detection of 1-vessel stenosis was 43% (6 of 14 patients), and for 2- and 3-vessel disease 79% (19 of 24) and 88% (16 of 18), respectively. Specificity was 92% (35 of 38), diagnostic accuracy 81%. The stenosed coronary artery was correctly localized in 85% of positive tests. Dipyridamole-induced increase in wall motion score index differed significantly between patients with 1-, 2-, and 3-vessel disease (0.02 +/- 0.17, 0.15 +/- 0.17, and 0.27 +/- 0.24, respectively), and early positive tests (dipyridamole dose of 0.56 mg/kg) were almost exclusively seen in patients with multivessel disease. Six patients (5%) developed symptomatic bradycardia and hypotension during the test. In conclusion, dipyridamole stress echocardiography is useful for detection and localization of coronary artery stenosis, particularly in patients with multivessel disease. PMID- 7499906 TI - Can dobutamine echocardiography distinguish necrotic from ischemic myocardium, early after myocardial infarction? AB - The aim of this study was to evaluate the usefulness of dobutamine echocardiography (DE) in distinguishing necrotic from ischemic myocardium in infarct zones. We performed DE in 39 patients, 3 to 5 days after admission for a first, acute myocardial infarction, treated with thrombolysis. DE was considered positive if wall motion in the infarct zone worsened progressively during increasing dose of dobutamine or if wall motion in the infarct zone initially improved at low dose of dobutamine and deteriorated at higher dose. The results of DE were correlated to the evolution of wall motion in the infarct zone after 3 months and to the need for supplementary balloon dilatation. In 15 of the 39 patients, there was evidence of residual ischemia in the infarct zone. Twenty of the 39 patients had a positive dobutamine echocardiogram. Eleven of these 20 patients had evidence of residual ischemia in the infarct zone. They showed generalized changes of wall motion in the total infarct territory during DE. The other 9 patients demonstrated only localized changes of wall motion in isolated segments of the infarct zone during DE. None of these patients had evidence of residual ischemia. IN CONCLUSION: DE seems worthwhile in the detection of residual ischemia in the region of infarction. To reduce the number of false positive DE early after myocardial infarction, only extensive changes of wall motion in the total infarct territory should be accepted as indicative of residual ischemia in the infarct zone. PMID- 7499907 TI - Transthoracic 2D echo color Doppler assessment of internal mammary artery to left anterior descending coronary artery graft. AB - For noninvasive evaluation of anatomy and flow characteristics of internal mammary artery graft (IMA-graft), 2D echo-Color-Doppler (CDE) was performed in 60 patients (54 M, 6 F, mean age 54.1 +/- 6.9 y), who underwent coronary angiography 20.1 +/- 13 months after a coronary artery bypass graft (CABG). CDE was performed, using an echocardiographic unit equipped with a 5 MHz linear transducer. In all patients, measurements of IMA-graft diameter (mm), and peak systolic and diastolic flow velocity (cm/sec) were obtained at baseline and also in 16 patients after dipyridamole infusion (0.54 mg/Kg/min) and in 10 patients after sublingual nitroglycerin (NTG) (0.4 mg). Angiography showed the IMA-graft patency in 58/60 patients (96.8%). A typical biphasic flow was displayed by CDE in 49/58 patients (84.4%) with angiographic patency. Dipyridamole infusion increased both IMA-graft diameter and peak diastolic flow velocity (PDFV) from 2.28 +/- 0.51mm to 2.9 +/- 0.42mm and from 19.4 +/- 6.2 cm/sec to 93.9 +/- 29 cm/sec, respectively (p < 0.0001). No significant modifications of peak systolic flow velocity (PSFV) were observed. NTG increased PDFV from 29.11 +/- 8 cm/sec to 41.88 +/- 7.20 cm/sec (p < 0.005), while diameter and PSFV showed no statistically significant modifications. CDE is a useful diagnostic tool for noninvasive evaluation of IMA-graft patency both early after surgery and during long-term follow-up. CDE pharmacological stress improves the sensibility of the technique and it can provide indirect information about pathophysiology of recipient coronary vessel. PMID- 7499908 TI - Significance of echocardiographic atrioventricular plane displacement for the evaluation of left ventricular filling and end-diastolic pressure in patients with coronary artery disease. AB - The aim of this study is to assess the left ventricular filling and estimate the end-diastolic pressure of the left ventricle in patients with coronary artery disease (CAD) by echocardiographic measurement of the atrioventricular plane displacement (AVPD). In 101 patients (mean age 59 +/- 12 years) with CAD, a complete transthoracic echocardiographic study was performed, just prior to cardiac catheterization. The AVPD was recorded by M-mode echocardiography, from apical four and two chamber views. The recordings were obtained at four sites, corresponding to the septal, lateral, anterior and inferior walls of the left ventricle. The mean AVPD resulting from atrial systole (At), the mean total (T) diastolic AVPD, the ratio At/T(%) and the ratio of mitral annulus excursion during early and late diastole [(T-At)/At] were calculated. Firty-two age-matched healthy subjects served as control group. Both At and At/T were significantly greater in patients with CAD than in the controls (6.06 +/- 0.94 vs 5.53 +/- 0.55 mm, p < 0.01 and 43.4 +/- 5.9% vs 33.49 +/- 4.45%, p < 0.001 respectively). The ratio [(T-At)/At] correlated with the E/A ratio of transmitral flow, both in healthy subjects (r = 0.850, p < 0.001) and in patients with CAD (r = 0.722, p < 0.001). Correlation also existed both in patients with segmental wall motion abnormality (SWMA) (r = 0.691, p < 0.001) and in patients with SWMA (r = 0.818, p < 0.001). In patients with CAD, At/T further correlated with the left ventricular end-diastolic pressure (r = 0.517, p < 0.001). In patients with SWMA and in patients without, a correlation was also found (r = 0.516, p < 0.001 and r = 0.566, p < 0.001 respectively).(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7499909 TI - Effect of pacing rate on regional left ventricular wall motion. Assessment by quantitative analysis of equilibrium radionuclide angiography. AB - We evaluated the hemodynamic impact of right ventricular pacing at different stimulation frequencies. Fourteen patients with a right ventricular pacemaker (VVI pacing with retrograde P wave) but without clinical and electrocardiographic evidence of coronary artery disease underwent two equilibrium radionuclide angiographies: one under low heart rate (50/60 beats per minute) and the other under fast heart rate (90/100 beats per minute). Left ventricular global and sectorial ejection fractions, amplitude and phase images of first harmonic, and sectorial phases of left ventricle were analyzed. In twelve patients (84.7%), sectorial ejection fraction abnormalities in the left ventricular apicoseptal and inferoapical regions were observed under low heart rate, and worsened under fast heart rate, while new onset sectorial ejection fraction abnormalities under fast heart rate were observed in the other two patients with normal sectorial ejection fraction under low heart rate. Sectorial ejection fractions of left ventricular apicoseptal and inferoapical regions significantly changed between low and high heart rate (- 14.1 +/- 3.8%, p < 0.005; - 7.5 +/- 2.4%, p < 0.01 respectively). Left ventricular sectorial phases were abnormal in only two patients (14.3%) under low heart rate, and in twelve patients (84.7%) under fast heart rate. Our study confirms that left ventricular regional wall motion abnormalities during VVI pacing significantly worsen under fast heart rate in comparison to those under low heart rate. PMID- 7499910 TI - Mobile right atrial mass biopsy guided by biplane transesophageal echocardiography. AB - Transesophageal echocardiography (TEE) has been successfully used in monitoring complex or high-risk interventions in the cardiac catheterisation laboratory. We report a case in which biplane TEE was used to facilitate the biopsy of a mobile right atrial mass. PMID- 7499911 TI - [Serotypes and antimicrobial susceptibility of Streptococcus pneumoniae from clinical specimens]. AB - Studies were made on 66 strains of Streptococcus pneumoniae which were obtained from clinical specimens in 1991 through 1993 and showed 19 mm or less of disk inhibition zone diameter (DIZD) against 1 microgram oxacillin (MPIPC) disk. The studies included the determination of their serotypes, antimicrobial susceptibility, and comparison between microbroth dilution (MD) method and Kirby Bauer (K-B) method. In the study of distribution of serotypes, additional 32 strains which showed 20 mm or more in DIZD were included for study. The results were as follows. 1) About 70% of 98 strains of S. pneumoniae were serotyped by 6 kinds of antisera. Among penicillin-susceptible S. pneumoniae (PSSP), type 3 were 20.6%, type 19, 15.9%, type 6, 14.3%, type 18, 9.5%, type 14, 7.9%, and type 4, 1.6%. Among penicillin-intermediate S. pneumoniae (PISP), and penicillin resistant S. pneumoniae (PRSP), type 19 were 60%, and type 18, 8.6%. In PISP and PRSP, more than half were type 19, which indicates they are distinctly different from PSSP in serotypical distribution. 2) As to the difference between screening by MPIPC disk and minimum inhibitory concentration (MIC) by benzylpenicillin (PCG), among 66 MPIPC resistant strains, PSSP strains were 31 in number (47%). 3) MIC showed that PISP and PRSP strains were more resistant than PSSP against cefaclor (CCL), cefazolin (CEZ), cefotiam (CTM), cefotaxime (CTX), imipenem (IPM), minocycline (MINO), and erythromycin (EM), but no difference was found in the 2 groups of strains in MIC with clindamycin (CLDM) and ofloxacin (OFLX). 4) All type 3 strains formed mucoid colonies and were resistant to MINO and highly resistant to EM and CLDM. 5) By NCCLS, category of antimicrobial susceptibility is determined against CCL, EM, OFLX, in MD method and K-B method. Against these antibiotics, the complete agreement rates were 75.8%, 92.4% and 86.4% respectively. PMID- 7499912 TI - [The use of polymerase chain reaction method for the detection of Rickettsia tsutsugamushi in wild rodents]. AB - We studied the applicability of polymerase chain reaction (PCR) method for the detection of R. tsutsugamushi in wild rodents. The PCR method which amplified the gene coding for the group-specific antigen of R. tsutsugamushi was used in this study. Specific PCR products (88 bp) were obtained with the DNAs from three reference strains (Gilliam, Karp, and Kato) and two cell culture adapted field isolates (KN-1 and GJ-1). The minimum number detectable by the PCR method was estimated to be 1.3 copies of rickettsial genome. In a study with experimentally infected mice, the PCR method could detect rickettsial DNA in one of two infected mice at four months after inoculation. Thereafter, fifty five wild rodents were captured in five areas of Okayama Prefecture, and R. tsutsugamushi DNA was detected, by the PCR method, by amplifying DNA from the spleen of each rodent. The rickettsia was also isolated from the same rodents by the mouse inoculation method. By the PCR method, rickettsia DNAs could be detected in 12 of 13 rodents from which the rickettsiae were isolated, and in 10 of 42 rodents from which no rickettsiae were isolated. These findings indicate that the PCR method is a simple and specific procedure to detect R. tsutsugamushi in wild rodents. On the other hand, the results of the PCR method demonstrated that the middle area of Okayama Prefecture was highly (44-81%) contaminated with R. tsutsugamushi. PMID- 7499913 TI - [Studies on tsutsugamushi disease in Gifu prefecture. 6. Correlation between number of patients and meteorological elements]. AB - The correlations between numbers of tsutsugamushi disease patients and meteorological elements were analyzed for 11 years from 1982 to 1992 in Gifu Prefecture, Japan by using regression analysis. The number of patients in early winter was closely correlated independently to both the mean of the minimum temperatures from 11th May to 31st July and the mean of the maximum temperatures in November in the same year. Regression coefficients (R2) were 0.689 and 0.560, respectively. On this basis, an equation for prediction of the number of patients in early winter was designed as follows: N = [e (j - 17.6) + 2.3(v - 13)] x j x v/156 (prediction formula 1) N:predicted number of patients in early winter j:the mean of the minimum temperature from 11th May to 31st July v: the mean of the maximum temperature in November e:the base of the natural logarithm (= 2.718...) The number of patients in early winter was also closely correlated to j in an equation of the fifth degree (R2 = 0.930). N = 22.524656384 j5 - 2218.23705 j4 + 87272.992 j3 - 1714734.329 j2 + 16825634.235 j - 65963810.254 Based on these formulas, the temperature in early summer has a significant effect upon the prevalence of tsutsugamushi disease in early winter. PMID- 7499914 TI - [Epidemiological study of tsutsugamushi disease in Gunma prefecture. A special field study and serotype]. AB - An epidemiologically investigated of invasion of Rickettsia tsutsugamushi, inhabitant of mites and serum sample from patients with Tsutsugamushi disease in Gunma prefecture from 1984 to 1994 was made. Our data clearly indicated that Rickettsia tsutsugamushi was not located but widely spreaded throughout the Prefecture. Mites on rodents, were classified into 4 genus and 12 species and about 15% of them were Leptotrombidium pallidum and Leptotrombidium scuttellare, well known virulent vectors. The highest incidence rate of this disease was observed in the northwest area of the Prefecture from October to December, while a smaller number of patients occurred in other areas and in other months. About fifty percent of the serum samples from the patients were positive to the Karp strain. These results suggest that the major cause of this disease is the Karp strain and the disease could occur potentially in various areas of the Prefecture. PMID- 7499915 TI - [Appearance of antibacterial activity of oxacillin against methicillin resistant Staphylococcus aureus (MRSA) in the presence of catechin]. AB - We previously reported that tea catechin shows bactericidal activity against various bacteria including methicillin resistant Staphylococcus aureus (MRSA) and that bactericidal catechin damages the lipid bilayer of bacterial cell membranes. Here we describe that oxacillin (MPIPC) shows antibacterial activity against MRSA in the presence of catechin below MIC. Twenty clinical isolates of MRSA were examined by a cup method. In the absence of catechin, MPIPC even at a concentration of 40 micrograms/ml did not show antibacterial activity against all isolates of MRSA. However, when catechin below MIC (25-100 micrograms/ml) was mixed with the agar medium, MPIPC (5-12.5 micrograms/ml) showed antibacterial activity against all MRSA isolates. By counting the numbers of viable bacteria in a broth culture, only MPIPC (5 micrograms/ml) or catechin (100 micrograms/ml) showed similar growth curves to the control. But addition of both MPIPC and catechin reduced the number of viable bacteria to 1/100-1/10000 after 24 hours of cultures. Besides MPIPC, in the presence of catechin below MIC methicillin (12.5 micrograms/ml), aminobenzylpenicillin (32 micrograms/ml), tetracycline (2.5 micrograms/ml), and chloramphenicol (12.5 micrograms/ml) showed antibacterial activities against multiple drug resistant MRSA to antibiotics mentioned above. These findings suggest a possible use of catechin in the treatment of MRSA infection. PMID- 7499916 TI - [Serological diagnosis for human parvovirus B19 infection by an enzyme immunoassay kit with recombinant antigens synthesized in a baculovirus expression system]. AB - Propagation of human parvovirus B19 (B19) in cell cultures are not applicable to the source of viral antigens for serological assays at present. Enzyme immunoassay (EIA) kits with recombinant B19 capsids by E. coli or baculovirus expression system have been developed. We tested serum samples from the patients with erythema infectiosum and aplastic crisis by EIA kit with recombinant antigens synthesized in a baculovirus expression system (Denka Seiken Co., Tokyo, Japan). The antigens used in the kit are self-assembled recombinants containing both VP-1 and VP-2 with the same proportion as found in native B19 capsids. B19 IgM is detected by antibody capture methods and IgG by indirect methods. All of the samples were positive for B19 DNA by nested PCR. Thirty-six (97%) of the 37 patients with erythema infectiosum and all (100%) of the 4 patients with aplastic crisis were positive for B19 IgM. The EIA kit with recombinant antigens synthesized in a baculovirus expression system has proved to be reliable and useful for the diagnosis of B19 infection. PMID- 7499918 TI - Micro-carrier-test: evaluating disinfectants for HIV. AB - To determine the effect of disinfectants against viruses in vitro, I devised the Micro-Carrier-Test of dry-fixed virus-infected cells. Human immunodeficiency virus type 1-infected Molt-4 cells (1 x 10(5) cells in 5 microliters of 10% fetal bovine serum) were dry-fixed at the bottom of each well of a 96-well flat bottomed microtiter plate for 120 minutes at room temperature. Disinfectants were added and allowed to remain for designated times and the wells were washed three times with PBS. Uninfected Molt-4 cells (1 x 10(4) cells/well) were inoculated and cultured for 4 weeks. The culture supernatant was harvested to measure reverse transcriptase activity by non-radioisotopic reverse transcriptase assay every week. Residual cytotoxicity of the disinfectant was determined by cytotoxicity assay. To evaluate the new method, the virucidal efficacy of several well-known disinfectants was reevaluated. Dose- and time-dependent effects of the disinfectants were determined. The minimal effective concentrations after 5 minutes of contact were 20% ethanol, 0.01% glutaraldehyde and 0.1% sodium hypochlorite. These results are almost the same as those reported previously, but there are some discrepancies. The differences between the present and previous protocols are discussed. This Micro-Carrier-Test promises to be useful in the screening of disinfectants. PMID- 7499917 TI - [Study on sepsis in the elderly at Nagoyashi-Koseiin Geriatric Hospital]. AB - A study based on clinical analysis was conducted regarding the 125 episodes in the elderly 112 patients of sepsis who were 70 (average 83.8 +/- 7.5) years old at Nagoyashi-Koseiin Geriatric Hospital from 1985 through 1994. 1) The backgrounds of the elderly patients with sepsis were as follows: bedridden (72.8%), urinary catheter in place (61.2%), central venous catheter in place (48.8%), and prior antibiotic use (40.8%). All patients had an underlying disease. 2) Organisms isolated were Escherichia coli (21.2%), Staphylococcus aureus (18.4%); Coagulase-negative staphylococci (CNS) (17.4%) and Candida albicans (6.1%). Chronologically, the quantity of gram-positive cocci increased while that of gram-negative bacilli decreased. As the age of the patients increased, the frequency of infections by Methicillin-resistant Staphylococcus aureus (MRSA), E. coli, and/or multiple bacteria increased, while that of infections by CNS and gram-negative bacilli excluding E. coli decreased. 3) The primary infected sites were the urinary tract system (24.8%), central venous catheter (21.6%) and unknown (31.2%). 4) The primary clinical observations were fever exceeding 38.0 degrees C (88.0%), tachycardia (60.8%), shivering (44.0%) and cyanosis (32.8%). 5) Complications were multiple organ failure (33.6%), septic shock (26.4%) and disseminated intravascular coagulation (22.4%). 6) The prognosis indicated that 65.6% were survivors, and 34.4% were nonsurvivors. At the onset of sepsis, weight, blood pressure, serum albumin, and total cholesterol in the nonsurvivors were significantly lower than those in the survivors, whereas heart rate, GOT, LDH, and BUN in the nonsurvivors were significantly higher than those in the survivors. PMID- 7499919 TI - [A case of Acanthamoeba keratitis after operation for cataract]. AB - Acanthamoeba keratitis occurs mainly in contact lens users. We experienced a patient with Acanthamoeba keratitis after operation for cataract. A 70-year-old male, who suffered from suppurative keratitis with impairment of visual acuity and eye pain in the left eye after the operation, was admitted to our hospital. After admission he received treatment with oral and topical antibiotics without any improvement. Neither bacterial or fungal pathogens was detected from corenal skrappings. Blue stained Acanthamoeba cysts were detected with the Parker ink KOH preparation from punctured fluid of the anterior chamber of the eye. Acanthamoeba cysts were also cultured on a nonnurient agar plate with Escherichia coli. Then he was treated with oral and topical miconazole and topical fluconazole. His visual acuity did not improve because of the lag of appropriate treatment. Therefore, attention must be paid for the existence of Acanthamoeba keratitis after ophthalmologic operations. PMID- 7499920 TI - [Isolation of Bilophila wadsworthia from peritoneal fluid of a patient with an infected Douglas pouch]. PMID- 7499921 TI - Skin lesions occurring in clients with reflex sympathetic dystrophy syndrome. PMID- 7499922 TI - Myasthenia gravis: pathophysiology, diagnosis and collaborative care. AB - Myasthenia Gravis (MG) is an autoimmune disorder which compromises neuromuscular transmission. The hallmark of the disorder is fatigue with repetitive activity. Patients may experience symptoms ranging from double vision, ptosis and weak voice to choking, shortness of breath, generalized weakness and respiratory failure. The clinical diagnosis is confirmed by identification of a decremental response to repetitive nerve stimulation by electromyography (EMG), the presence of serum antibodies to the muscle acetylcholine receptor (AChR), or an improvement in strength with administration of intravenous edrophonium. With improvements in critical care and immunosuppressive treatments, MG is rarely the grave disease it once was, but because of the odd fatiguing symptoms and relative rarity of the disorder, patients are frequently misdiagnosed and their special needs overlooked. The nature of MG, with its acute and chronic components, creates complex needs for affected individuals and their families. The advanced practice nurse in collaboration with a neurologist in the outpatient setting is positioned to address these needs in an ongoing case management role. PMID- 7499923 TI - Functional Assessment of Cancer Therapy Scale. PMID- 7499924 TI - Karnofsky performance scale. PMID- 7499925 TI - Multidimensional Quality of Life-Cancer Version 2. PMID- 7499926 TI - Quality of life from the patient's perspective. PMID- 7499927 TI - Deep vein thrombosis and pulmonary emboli in neuroscience patients. AB - Patients with neurologic dysfunction, particularly spinal cord-injured and stroke patients, are at risk for developing thromboembolic complications due to a variety of factors, especially impaired mobility. Venous stasis, altered coagulability and endothelial vessel damage create a triad of pathophysiological changes that place patients at risk for developing deep vein thrombosis. Prophylactic use of pneumatic compression boots is highly recommended while administration of anticoagulants remains controversial and varied. Inferior vena cava filters are placed in patients who are high risk for pulmonary emboli. Nursing interventions focus on assessments of risk potential and clinical signs and symptoms and implementation of prevention strategies. PMID- 7499928 TI - Health maintenance behaviors in advanced Parkinson's disease. AB - Co-morbidity in the older adult Parkinson's disease patient decreases functional status and quality of life, and increases the risk of permanent disability. Although many chronic conditions in the adult population can be prevented, or at least influenced by early detection and treatment many progressive neurologic disorders such as Parkinson's disease cannot. This pilot study describes the health maintenance and disease prevention behaviors of a group of patients with Parkinson's disease. Findings demonstrate that parkinsonians attend in a way similar to their peers without Parkinson's disease to most parameters of general health maintenance as established by the United States Preventive Task Force. In fact, in 11 out of 12 selective parameters, parkinsonians exhibit increased frequency of participation in the recommended behaviors. However, achievement of our national goals for health promotion and disease prevention will require additional efforts in this population. Emphasis should include the participation in Pap smears, mammograms, exercise, seat belt usage and adult immunizations, especially the pneumococcal vaccination. Neuroscience nurses can play an important role in reaching health promotion and disease prevention goals in persons with Parkinson's disease through patient education, advocacy, case management, anticipatory guidance and referrals to primary care providers. PMID- 7499929 TI - An acute spinal cord injury: my family's story. AB - An acute spinal cord injury has been described as one of the most devastating traumatic types of neurological impairment. Twelve years ago I had the misfortune of experiencing this first hand. From the day of my brother's initial injury I began keeping a diary of what this experience was like. The concept of entitlement and indebtedness has been applied to my family's experience. PMID- 7499930 TI - Perceptual accuracy and gender-related differences in parents of children with myelomeningocele. AB - The ability to perceive accurately is central to efficient and optimal use of health care resources. Many factors may affect parental ability to achieve this accuracy including concepts pertinent to perception theory and gender of the parent. Facets of person perception theory, gender-related differences between parents and suggestions for nursing care are discussed in the context of caring for a child with myelomeningocele. PMID- 7499931 TI - Home epidural analgesic infusion for non-cancer pain. AB - Continuous epidural analgesic infusion is widely recognized to be helpful for pain relief in the patient with cancer pain, but it can be as beneficial for the patient with non-cancer pain. Cancer pain patients have been offered long-term epidural infusion for relief of chronic pain, but non-cancer pain patients have less frequently been provided the same opportunities for pain relief. Both patient outcome and cost-effectiveness must dictate treatments offered as alternatives to long-term use of oral or injectables, nonsteroidal anti inflammatories and muscle relaxants. It is necessary for nurses practicing in pain management and home health settings to understand the anatomy, neuropharmacology and nursing implications in order to effectively care for this growing population of patients. Epidural infusion is a safe and effective route of pain relief if the medical criteria as been met for choosing the patients receiving the treatment, and if the caregivers understand the treatment and expected outcomes. This article discusses anatomic considerations in drug selection, physiology, patient selection, neuropharmacology and patient care for epidural infusion. This article is designed to assist the nurse in accomplishing the patient and physician goals in this treatment modality for chronic non-cancer pain. PMID- 7499932 TI - Monitoring neuromuscular function. PMID- 7499933 TI - A tangle of feelings. PMID- 7499934 TI - Squamous cell carcinoma in thyroglossal duct cyst. AB - Although a thyroglossal duct cyst (TDC) is a common cyst occurring in the neck, carcinomas arising in the TDC are a rare event. To date, approximately 100 cases have been reported, the majority of them being papillary carcinomas. Squamous cell carcinomas (SCC) are very rare, and only nine cases have been reported so far in the literature. We present a 55-year-old female patient with a SCC in a TDC to highlight the clinical and pathological features of this condition. PMID- 7499935 TI - Atlanto-axial subluxation (Grisel's syndrome) following otolaryngological diseases and procedures. AB - Grisel's syndrome is a rare condition of uncertain aetiology characterized by atlanto-axial subluxation following an infection in the head and neck region. The condition most frequently affects children and characteristically presents in the post-operative period with cervical pain and torticollis. We describe three cases and discuss the aetiology and pathogenesis. A high index of suspicion may lead to early diagnosis in its natural history thereby preventing potentially serious complications. PMID- 7499937 TI - Ramsay Hunt syndrome mimicking acoustic neuroma on MRI. PMID- 7499936 TI - Facial palsy due to tuberculosis: the value of CT. AB - This case report demonstrates the value of CT in the investigation of facial palsy due to tuberculous mastoiditis. This is the best imaging modality available for identifying features suggestive of tuberculosis, the disease extent, presence of complications and demonstration of anatomy prior to surgery. PMID- 7499938 TI - Schwannoma of the external auditory canal. AB - Solitary schwannomas are relatively rare. There has been only one previously reported case, in the English literature, of a schwannoma arising in the external auditory canal. We present a case which was managed by local excision and was unassociated with cochlear or retrocochlear pathology. PMID- 7499939 TI - Ameloblastic carcinoma of the maxilla. AB - Ameloblastic carcinoma is an unusual tumour. There have been a total of 34 cases of ameloblastic carcinoma in the English literature to date. Of these only 11 cases have occurred in the maxilla. The authors report the 12th such case. The histological classification for odontogenic carcinoma has been debated for many years and recently revised, thus differentiating between malignant ameloblastoma and ameloblastic carcinoma. The authors review the current literature regarding diagnosis and treatment of this unusual lesion, and support the use of the term malignant ameloblastoma for the tumours that metastasize in spite of their benign histological appearance, whereas, the ameloblastic carcinoma is referred to as the primary tumour with malignant transformation, regardless of its metastatic potential. PMID- 7499941 TI - Tympanosclerosis and mini grommets: the relevance of grommet design. AB - Fifty children with otitis media with effusion undergoing grommet insertion had into one ear a Mini-titanium grommet inserted and into the other ear a Mini teflon grommet. Post-operative follow-up until after extrusion of the grommets demonstrated only a small difference between the extrusion times of the two grommets (a significant difference of 41 days) and no difference in the degree of tympanosclerosis seen with each grommet. We propose that the mass of a grommet appears to play less of a role than has previously been suggested in the pathogenesis of tympanosclerosis following grommet insertion and that duration of intubation may be the most significant factor. PMID- 7499940 TI - Is central hyperacusis a symptom of 5-hydroxytryptamine (5-HT) dysfunction? AB - The subjective symptom of hyperacusis is described. The terms 'hyperacusis' and 'phonophobia' are considered to be synonymous as there is no recognized distinction between these two descriptions. Peripheral auditory pathologies with associated hearing sensitivity are reviewed and the likely mechanisms underlying the hyperacusis are listed. The neurological conditions, which have been reported to occur with hyperacusis, are reviewed. A separate aetiology of central hyperacusis is therefore proposed, with a symptom profile distinct from the peripheral hyperacusis. A common factor to neurological conditions with hyperacusis, is disturbance of 5-HT (5-hydroxytryptamine) or serotonin function. The research literature on the role of 5-HT in sensory modulation (specifically auditory startle) in animals is presented. It is proposed that 5-HT dysfunction is a probable cause of increased auditory sensitivity manifested as central hyperacusis or phonophobia. PMID- 7499942 TI - Cisplatin in children: hearing loss correlates with iris and skin pigmentation. AB - Pigmentation is reported to affect cisplatin-induced ototoxicity in adult humans. The hearing loss is worse in people with brown irises, than in those with blue irises. We assessed the hypothesis that cisplatin-treated children with dark irises suffer more deterioration in auditory thresholds than do those with less pigmentation. For the 19 children whose data met the requirements of this observational retrospective study, we found a weak correlation (Spearman's r = 0.50; p < 0.05) of high frequency hearing loss (at 4000 Hz) and pigmentation. Blue or hazel-eyed children averaged 2.9 dB worsening at 4000 Hz, in contrast to 14.2 dB worsening for brown or black-eyed children. Pigmentation may account for some of the individual susceptibility to cisplatin ototoxicity. We suggest that iris colour be included in future reports of cisplatin-related hearing loss. PMID- 7499943 TI - Hearing impairment in association with distal renal tubular acidosis among Saudi children. AB - A follow-up of seven patients with the autosomal recessive inherited syndrome of distal renal tubular acidosis (RTA) and sensorineural hearing loss is described. Five patients were diagnosed as having primary distal renal tubular acidosis and rickets, four were found to have severe sensorineural hearing loss of over 80 dB: two of which are brothers. Two patients were diagnosed as having secondary distal renal acidosis due to a genetic disorder called osteopetrosis; they are brothers and their audiograms showed a mild conductive hearing loss of an average 35 dB bilaterally. All patients had growth retardation with improvement due to alkaline therapy but their hearing loss was not affected by the medication. The pedigrees of two families with half sibs showed the familial incidence for consanguineous marriage. Consanguinity was found to be positive in five out of the seven patients. The tribal tradition in Saudi Arabia fosters consanguineous marriages for cultural and social reasons and pre-arranged marriages are still seen. PMID- 7499944 TI - The use of a CT scan to predict the feasibility of decompression of the first segment of the facial nerve via the transattical approach. AB - Total surgical decompression of the facial nerve can be offered to patients severely affected by Bell's palsy whether via the transattical or middle fossa approach. We prefer, when feasible, the transattical approach because it does not violate the cranial vault. The purpose of this study was to find the anatomical parameter of the temporal bone, measured by means of computed tomography (CT scan) and to decide which of these approaches should be offered. Sixty temporal bones were studied by CT scan, and then dissected in order to perform total facial nerve decompression via the transattical approach. Correlation between the two studies was established. Results suggest that measurement by CT scan of the attical area in the axial plane (AAA) may determine those patients for whom the transattical approach to facial nerve decompression should be undertaken. PMID- 7499945 TI - Effect of Beconase nasal spray on olfactory function in post-nasal polypectomy patients: a prospective controlled trial. AB - Numerous studies have postulated the possible benefit of corticosteroids on olfaction in patients with nasal/sinus disease. Twenty-nine patients with bilateral nasal polyps were included in our study using strict selection criteria to reduce other aetiologies of olfactory dysfunction. The University of Pennsylvania Smell Identification Test (UPSIT) was performed pre-operatively on the right and left nostrils separately. Following intranasal polypectomy the patients received a six-week course of beclomethasone nasal spray (Beconase) to one nostril only, with the other acting as a control. The UPSIT scores were again obtained for each nostril separately. Wilcoxon Signed Rank test revealed no statistically significant difference in UPSIT scores between treated and untreated nostrils (p = 0.31; power 70 per cent; ES = 0.47). We conclude that topical beclomethasone does not improve olfaction following nasal polypectomy. PMID- 7499946 TI - Sinogenic intracranial complications. AB - Two hundred and nineteen patients, with intracranial complications of sinusitis, are presented. Sinusitis is still a life-threatening condition and if neglected, or mismanaged, can lead to intracranial complications that result in a high mortality and morbidity. Twenty-two patients had meningitis, 127 subdural empyema, 38 brain abscess, 15 combined brain abscess and subdural empyema and 17 extradural empyema. The diagnosis of intracranial abscess and sinusitis was made with the aid of a CT scan, and that of meningitis on cerebrospinal fluid microscopy, chemistry and culture. The most frequent presenting signs were fever (68 per cent) and headache (54 per cent). The most common localizing neurological sign was hemiparesis (35.5 per cent). Orbital inflammation was present in 41.5 per cent of patients. Treatment entailed immediate, appropriate, intravenous antibiotic therapy and prompt surgery, performed within 12 hours of admission. In patients with meningitis, the surgery entailed surgery of the sinus disease only. In patients with subdural empyema, brain abscess and extradural empyema, evacuation of the primary source of infection by the radical frontoethmoidectomy approach, immediately after drainage of the intracranial collection of pus, was carried out. There were 35 deaths (16 per cent). The highest mortality rate was recorded in patients with meningitis (45 per cent) followed by brain abscess (19 per cent) and subdural empyema (11 per cent). Despite advances in medicine, i.e. antibiotics and CT scan for early and accurate diagnosis, the mortality from sinogenic intracranial complications has remained significant. This can only be eliminated through education.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7499947 TI - Endoscopic anatomy of the sphenoid sinus and sella turcica. AB - An endoscopic study of the sphenoid sinus was carried out, on 30 cadavers, to understand the important anatomical relationships of the sphenoid sinus, and the sella turcica. The aim was to study the endoscopic anatomy and the variants, and to determine if endoscopic instrumentation and techniques, could play a beneficial role in endoscopic management of sellar lesions. The results of this study are discussed, with particular reference to the important surgical anatomical features of the sphenoid sinus. A surgical technique for the endoscopic transsphenoid approach to the sella turcica was developed. Anatomical variants can be identified endoscopically, and endoscopic techniques have the advantages of improved visualization, magnification, angled vision, and a panoramic perspective of the intrasphenoid anatomy, compared to currently employed methods of pituitary/sellar surgery, using the operating microscope. PMID- 7499948 TI - Endoscopic management of lesions of the sella turcica. AB - The excellent visualization and minimally invasive surgical technique of endoscopic sinus surgery was applied to the management of 40 patients with sellar lesions. Endoscopic management of sellar lesions offers, not only the advantage of improved visualization, but also magnification, and a panoramic perspective of the important relationships of the sella turcica. In the past year, we have managed 40 subjects with sellar lesions, endoscopically: 38 patients had pituitary adenomas and two a craniopharyngioma. At our hospital, the endoscope has replaced the operating microscope for surgery for pituitary adenomas and other sellar lesions. The endoscopic approach to the sphenoid sinus and the sella is performed by an ENT surgeon and the ablative surgery performed by a neurosurgeon. Our experiences, using the endoscope to perform surgery on sellar and parasellar lesions, are reported and the advantages, over the operating microscope, which is traditionally used are discussed. The technique for endoscopic management of sellar lesions is described. PMID- 7499949 TI - Post-tonsillectomy pain: the first ten days. AB - Forty-one adults were entered into a prospective study to investigate the pattern of pain over 10 days after elective dissection tonsillectomy. The severity of pain was scored by the patient on a visual analogue scale. The results confirm that tonsillectomy is a very painful experience and the provision of adequate analgesia is of paramount importance, especially if day-case tonsillectomy is performed. PMID- 7499950 TI - Ingested foreign bodies--a contemporary management approach. AB - A prospective study of all foreign body complaints presenting through our Accident and Emergency Department was conducted in a population where the condition is endemic. All patients were managed by otolaryngologists. Six hundred and eight patients were attended to yielding 179 foreign bodies. Making use of modern equipment and a practical approach, the requirement for examination under general anesthesia was 6.3 per cent. In this series there was a complication rate of 0.5 per cent. PMID- 7499951 TI - Is a community-based mastoid microsuction service feasible: the audit of a pilot project. AB - Patients who have had a mastoidectomy form a considerable long-term follow-up commitment to their local ENT Department. A community-based mastoid aural toilet pilot project run by a GP is described using an operating microscope and suction apparatus in his surgery. To obtain the necessary information about community treatment, an audit was performed of all mastoidectomy patients from a large general practice in Portsmouth: 57 mastoidectomy patients were reviewed. During the project it was possible to transfer to the community microsuction project the long-term care of most of their mastoidectomy patients who were attending the hospital ENT outpatient clinic. PMID- 7499953 TI - Cochlear reimplantation. AB - Since its inception in 1988 the Cochlear Implant Programme in Manchester has successfully implanted 69 adults and 23 children. Of these 92 procedures, three patients have undergone revision surgery with the insertion of either a new implant or re-positioning of the existing device. We examine the circumstances that lead to the need for reimplantation in these patients, discuss the technical aspects of revision surgery together with the functional results of such procedures. PMID- 7499952 TI - Haemangiopericytoma of the middle ear: case report and literature review. AB - Haemangiopericytoma is an uncommon vascular tumour that rarely appears in the head and neck region. Such a tumour arising from the middle ear has not been reported since it was first described in 1942 by Stout and Murray. The present case therefore illustrates the first example of a primary middle ear haemangiopericytoma. PMID- 7499954 TI - Renal cell carcinoma presenting as a solitary paranasal sinus metastasis. AB - We present an unusual case of renal cell carcinoma presenting as a periorbital mass. The presenting symptom was a supraorbital swelling which proved to be a mucocoele which had arisen secondary to a solitary metastasis in the ethmoidal sinuses. We discuss the management strategy including the place of endoscopic biopsy, the choice of investigations and craniofacial resection. PMID- 7499955 TI - Migrating foreign body: a new cause of trismus. AB - A unique case of delayed trismus secondary to an infratemporal foreign body is presented. The pathophysiology and treatment options are discussed. PMID- 7499956 TI - A case of submandibular teratoma. PMID- 7499957 TI - Malignant myoepithelioma of the parotid gland: case report and review of the literature. AB - A 62-year-old male with a myoepithelioma of the right parotid gland was treated with surgical excision followed by adjuvant radiation therapy. Prior to the completion of radiation therapy, the patient developed progressive disease at local, regional, and distant metastatic sites. Combined modality treatment with radiation and chemotherapy resulted in a significant but transient shrinkage of the tumours at all sites. The patient succumbed to metastatic disease 212 days following the diagnostic biopsy. This case illustrates several of the distinctive clinical and pathological characteristics of this rare tumour. PMID- 7499958 TI - A combined approach excision of branchial fistula. AB - Second branchial cleft and pouch anomalies are by far the commonest of the branchial anomalies. In most cases, however, there is a unilateral cleft sinus. A complete fistula with external and internal openings is rare. We present a case with a complete branchial fistula in which the tract was followed from the neck to the tonsillar fossa using a combined approach. PMID- 7499959 TI - Diverse immunological and hematological effects of interleukin 7: implications for clinical application. AB - Interleukin-7 (IL-7) was originally discovered to be a pre-B cell growth factor. Soon thereafter, a broader role for IL-7 in leukocyte development and function began to be identified. IL-7 now has been shown to be a critical cytokine for normal T and B lymphopoiesis and a mobilizer of pluripotent stem cells and myeloid progenitors. IL-7 has been demonstrated to enhance T cell function and induce cytokine expression in monocytes. Preclinical studies have already found that IL-7 could accelerate murine lymphocyte regeneration following chemotherapy and bone marrow transplantation, induce antitumor effects in mice, and expand anti-HIV-specific human T cells. Thus it is essential that further preclinical and clinical research be performed to evaluate IL-7 as a potential therapy for leukopenia, bone marrow/stem cell transplantation, cancer, and HIV/AIDS. PMID- 7499961 TI - On the expression of nitric oxide synthase by human macrophages. Why no NO? AB - The production of nitric oxide (NO) and its role in the anti-tumor and anti microbial effects of rodent macrophages appears well established. In contrast, the circumstances required for its release from human monocytes/macrophages and its potential role in human pathology remain controversial. Evidence to be discussed suggests that NO is a redundant, autotoxic, immunosuppressive, and inefficient mediator of macrophage function. For these reasons, the expression of nitric oxide synthase as a rapid-response, high-output effector pathway may have been evolved out of the human monocyte/macrophage response repertoire or severely restricted in its expression. Hypothetical roles for a modest and circumscribed production of NO by human macrophages are proposed. PMID- 7499960 TI - The effect of granulocyte-macrophage colony-stimulating factor on myeloid cells and its clinical applications. AB - Granulocyte-macrophage colony-stimulating factor (GM-CSF) is a naturally occurring growth factor produced by several cell types in response to a variety of stimuli. GM-CSF has potent stimulatory effects on the growth and maturation of hematopoietic cells and has profound effects on mature circulating effector cells. Clinical applications of GM-CSF include ameliorating chemotherapy-induced neutropenia and enhancing hematopoietic recovery after bone marrow transplantation. This review evaluates the effect of GM-CSF on myeloid cells and its clinical applications. PMID- 7499962 TI - Induction of apoptotic cell death in mouse lymphoma and human leukemia cell lines by a calcium-binding protein complex, calprotectin, derived from inflammatory peritoneal exudate cells. AB - We have previously shown that the calcium-binding protein complex, calprotectin, purified from rat inflammatory peritoneal cells exerts marked cytotoxic activity against rat, mouse, and human tumor cells. We studied here whether the cytotoxicity is caused by induction of apoptosis, using mouse EL-4 lymphoma and human MOLT-4 leukemia lines as targets. The rat calprotectin sample inhibited [3H]thymidine incorporation into these cells by partially 24 h and almost completely in 48 h of culture at concentrations of 100-200 micrograms/ml. Morphological changes, that is, loss of cell volume and nuclear condensation and/or fragmentation, appeared in both cell types cultured with calprotectin from 20 h, and such apoptotic cells subsequently increased in number to compose the great majority of the cells at 40 h. Cell death, measured by stainability with trypan blue, lagged behind the emergence of the apoptotic morphology by about 2 and 10 h in EL-4 and MOLT-4 cells, respectively. DNA fragmentation was observed in EL-4 cells cultured with calprotectin, whereas it was not observed in MOLT-4 cells, consistent with results of flow cytometry showing that loss of cell DNA content caused by the factor was greater in EL-4 cells. The data indicate that calprotectin induces the apoptosis of certain tumor cells but that the occurrence of DNA fragmentation is dependent on cell type. Finally, the apoptosis-inducing activity of the calprotectin sample was abrogated by the presence of 10 microM zinc, whereas it was not affected by 5 mM calcium or magnesium. PMID- 7499963 TI - Stability of nuclear segments in human neutrophils and evidence against a role for microfilaments or microtubules in their genesis during differentiation of HL60 myelocytes. AB - The nucleus of the mature human neutrophil is segmented into three to five interconnected lobes. The physiological purpose of this segmentation is unknown, as is the mechanism by which the lobes are formed during differentiation. Using video observation of migrating human neutrophils simultaneously illuminated for fluorescence and phase-contrast microscopy, we analyzed nuclear movements with respect to cell shape changes. The number of nuclear lobes and their relative size remained constant during observation (up to 1 h). The thin connecting segments between the lobes elongated and attenuated extensively but never separated. Electron microscopic analysis of neutrophil nuclei revealed no specialized nuclear or cytoplasmic structures in the vicinity of connecting segments. With fluorescence in situ hybridization of whole chromosome probes, we determined that chromosomes are randomly distributed among neutrophil nuclear lobes. HL60 cells are a human myelocytic line that, with retinoic acid treatment, segment their nuclei and differentiate into neutrophil-like cells over several days. Using a rapidly responding variant line termed HL60/S4 (Cancer Res. 52, 949 954), we found that segmentation could be induced within 24 h. We tested the role of cytoskeletal elements in the process of nuclear segmentation. Neither the microtubule inhibitor nocodazole nor the microfilament inhibitor cytochalasin D prevented nuclear segmentation. Together, our studies suggest that nuclear lobes in neutrophils are relatively stable structures that are not generated by microtubule- or microfilament-dependent forces. PMID- 7499964 TI - Taurine chloramine, a product of activated neutrophils, inhibits in vitro the generation of nitric oxide and other macrophage inflammatory mediators. AB - Taurine (Tau) is an exceptionally abundant free amino acid in the cytosol of inflammatory cells and especially in neutrophils. Taurine protects cells from self-destruction during processes that generate oxidants. The major function of Tau in leukocytes is to trap chlorinated oxidants (HOCl). Taurine reacts with HOCl to produce the long-lived compound taurine chloramine (TauCl). Previously, we have shown that other products of the neutrophil chlorinating system are able to modify functions of macrophages. In this study, we investigated in vitro the influence of TauCl on the generation of inflammatory mediators by activated macrophages. We have found that TauCl inhibited the generation of nitric oxide, prostaglandin E2, tumor necrosis factor alpha, and interleukin-6, but TauCl slightly enhanced the release of IL-1 alpha. The formation of nitrites by interferon-gamma-activated macrophages was inhibited by TauCl in a dose-dependent manner. Taurine chloramine also reduced the level of inducible nitric oxide synthase (iNOS) mRNA in macrophages, in a similar concentration-dependent manner. Although our experiments do not exclude a direct effect of TauCl on enzymatic activity of iNOS, the inhibition of iNOS expression seems to be the major mechanism responsible for suppression of NO formation. Finally, we discuss the biological role of TauCl in vivo. We suggest that at the site of inflammation TauCl works as a specific signaling molecule of activated neutrophils that coordinates the generation of inflammatory mediators in macrophages. PMID- 7499965 TI - An analogue of lipid A and LPS from Rhodobacter sphaeroides inhibits neutrophil responses to LPS by blocking receptor recognition of LPS and by depleting LPS binding protein in plasma. AB - When incubated with lipopolysaccharide (LPS) in the presence of plasma, neutrophils become primed for enhanced release of superoxide in response to triggering by formyl-Met-Leu-Phe (fMLP). The effect of LPS on phagocytes is inhibited by a synthetic lipid A precursor, LA-14-PP (lipid IVa) or by LPS from Rhodobacter sphaeroides (Rs). We studied the mechanisms by which LA-14-PP or Rs LPS inhibited LPS-induced responses. When neutrophils were exposed to LA-14-PP or Rs-LPS for 3 min and then to Escherichia coli-LPS, the antagonists inhibited priming for superoxide release, and also blocked up-regulation of CD11b and adherence. This inhibition was dependent on plasma, was not overcome by higher amounts of E. coli-LPS or plasma, and was not observed at 0 degrees C, suggesting that E. coli-LPS was not able to interact with its receptor or other cellular recognition molecule in neutrophils that had been exposed to the antagonists. The alternative possibility that LA-14-PP or Rs-LPS depleted a plasma cofactor, resulting in inhibition of priming, was investigated by using LPS from Porphyromonas gingivalis (Pg) and Bordetella pertussis (Bp). These LPS primed neutrophils in a plasma-dependent and CD14-dependent manner, but were not blocked by LA-14-PP or Rs-LPS. When sub-optimal concentrations of plasma were exposed to LA-14-PP or Rs-LPS, and then mixed with Pg-LPS or Bp-LPS, followed by incubation with neutrophils, priming and up-regulation of CD11b were inhibited, and this inhibition was overcome by increasing the concentration of plasma. Binding of LPS binding protein (LBP) in plasma to immobilized E. coli-LPS was inhibited by pre incubation of plasma with LA-14-PP or Rs-LPS. Together with the result that treatment of plasma with anti-LBP antibody abolished the cofactor activity of plasma, these results indicated that LA-14-PP and Rs-LPS depleted LBP from plasma, resulting in inability of LPS to act on neutrophils. Thus LA-14-PP and Rs LPS inhibited the action of LPS on neutrophils by at least two mechanisms, blocking of LPS receptor recognition and depletion of the cofactor LBP. PMID- 7499966 TI - Not only Th2 cells but also Th1 and Th0 cells express CD30 after activation. AB - To investigate whether the CD30 molecule, expressed only by a minority of T and B cells, defines a subtype of T helper cells, Pityrosporum orbiculare-specific CD4+ T cell clones were assessed for CD30 protein and gene expression. The clones were defined as Th1, Th0, and Th2 according to their cytokine mRNA profile detected by reverse transcription PCR (RT-PCR). The kinetics of CD30 expression after OKT3 (anti-CD3) stimulation was analyzed by flow cytometry, immunocytochemistry, and RT-PCR. OKT3 activation induced a high expression of CD30 in cells of both Th1 and Th0 as well as Th2 type after 1-3 days. A difference between the clones was noted in that the Th2 clones remained highly positive in CD30 expression, whereas expression in the other clones started to decline from day 3. These data indicate that CD30 is expressed in activated CD4+ T cells of all three subtypes, and that the expression is sustained in Th2 cells. PMID- 7499967 TI - Cross-linking of CD18 primes human neutrophils for activation of the respiratory burst in response to specific stimuli: implications for adhesion-dependent physiological responses in neutrophils. AB - Adhesion is known to prime neutrophils for physiological activation in response to cytokines and other stimuli. We have employed the technique of receptor cross linking to study the potential role of CD18, the common beta-subunit of the beta 2-integrin family of adhesion molecules, in the regulation of the respiratory burst, as measured by luminol-enhanced chemiluminescence and iodination, in human neutrophils. CD18 cross-linking primed neutrophils to activate the respiratory burst after stimulation with tumor necrosis factor alpha (TNF-alpha) (100 units/mL), formylmethionyl-leucyl-phenylalanine (fMLP) (1 microM), and granulocyte-macrophage colony-stimulating factor (GM-CSF) (1 micrograms/mL), but not granulocyte colony-stimulating factor (G-CSF) (1 micrograms/mL), interferon gamma (IFN-gamma) (100 U/mL), or phorbol myristate acetate (100 nM). The maximal rate of chemiluminescence induced by fMLP, TNF-alpha, and GM-CSF was enhanced 8-, 6-, and 1.5-fold, respectively, following CD18 cross-linking. Priming of the respiratory burst by direct engagement of CD18 was confirmed in neutrophil mediated iodination experiments, where iodination induced by TNF-alpha, fMLP, and GM-CSF was increased 15-, 20-, and 7-fold, respectively, by CD18 cross-linking. Immunoblot experiments demonstrated that TNF-alpha-induced tyrosine phosphorylation was both accelerated and more intense in neutrophils after cross linking of CD18. Major tyrosine phosphoprotein products include proteins with approximate molecular masses of 40, 70, and 110 kDa. Genistein (50 microM), a selective tyrosine kinase inhibitor, reduced the TNF-alpha-stimulated respiratory burst by > 80% whether or not CD18 was cross-linked. These results affirm the importance of CD18 in adhesion-dependent priming of neutrophil functions and demonstrate that CD18 engagement per se is sufficient to prime neutrophils for cytokine-induced signal transduction mediated by tyrosine phosphorylation. PMID- 7499968 TI - Vitamin E supplementation with interferon-gamma administration retards immune dysfunction during murine retrovirus infection. AB - Murine retrovirus infection induces loss of vitamin E and immune dysfunction with loss of cytokine production by T-helper cells. Therefore interferon-gamma (IFN gamma) was given during dietary vitamin E supplementation to effectively prevent murine retrovirus-induced immunosuppression, cytokine dysregulation, and development of murine AIDS. Administration of IFN-gamma during vitamin E supplementation significantly prevented development of retrovirus-induced suppression of splenic natural killer cell activity and T cell proliferation. It also significantly slowed retrovirus-induced elevation of T helper (Th) 2 cytokine [interleukin (IL)-4, IL-5, and IL-10] production and monokine (IL-6 and tumor necrosis factor-alpha) secretion by splenocytes. The treatment also prevented loss of Th1 cytokine (IL-2 and IFN-gamma) secretion by splenocytes from retrovirus-infected mice alleviating splenomegaly and hypergammaglobulinemia. The combined therapy had an additive therapeutic impact. It was more effective than IFN-gamma treatment or vitamin E supplementation alone in delaying the development of retrovirus-induced immunosuppression with its cytokine dysregulation. PMID- 7499970 TI - Interferon-gamma inhibits HIV-induced invasiveness of monocytes. AB - HIV-infected monocytes form highly invasive network on basement membrane matrix and secrete high levels of 92-kd metalloproteinase (MMP-9), an enzyme that degrades basement membrane proteins. In the present study, using matrigel as a model basement membrane system, we demonstrate that treatment of human immunodeficiency virus (HIV)-infected monocytes with interferon-gamma at 50 U/ml inhibited the ability of infected monocytes to form an invasive network on matrigel and their invasion through the matrigel matrix. These effects were associated with a significant reduction in the levels of MMP-9 produced by HIV infected monocytes treated with interferon-gamma 1 day prior to infection with HIV as compared with that of untreated HIV-infected monocytes. Monocytes treated with interferon-gamma 1 day after HIV infection showed the presence of integrated HIV sequences; however, the levels of MMP-9 were substantially lower than those produced by monocytes inoculated with live HIV, heat-inactivated HIV, or even the control uninfected monocytes. Exposure of monocytes to heat-inactivated HIV did not result in increased invasiveness or high MMP-9 production, suggesting that regulation of metalloproteinase by monocytes was independent of CD4-gp120 interactions and required active virus infection. Furthermore, addition of interferon-gamma to monocytes on day 10 after infection inhibited MMP-9 production by more than threefold with no significant reduction of virus replication. These results indicate that the mechanism of interferon-gamma induced down-regulation of MMP-9 levels and reduced monocyte invasiveness may be mediated by a mechanism independent of antiviral activity of IFN-gamma in monocytes. Down-regulation of MMP-9 in HIV-infected monocytes by interferon-gamma may play an important role in the control of HIV pathogenesis. PMID- 7499969 TI - The role of tumor necrosis factor, interferon-gamma, transforming growth factor beta, and nitric oxide in the expression of immunosuppressive functions of splenic macrophages induced by Mycobacterium avium complex infection. AB - In order to verify the participation of some cytokines in the expression of the suppressor activity of splenic macrophages (M phi s) induced by Mycobacterium avium complex (MAC) infection, we studied whether anticytokine antibodies were capable of blocking their suppressor activity against concanavalin A (ConA) induced mitogenesis of splenocytes (SPCs). When either anti-tumor necrosis factor (TNF), anti-transforming growth factor-beta (TGF-beta), or anti-interferon-gamma (IFN-gamma) antibody was added to culture medium, suppressor activity was markedly reduced, in the order of anti-TNF, anti-IFN-gamma, and anti-TGF-beta antibodies. By contrast, neither anti-interleukin-6 (IL-6) nor anti-IL-10 antibody exerted such a blocking effect. Therefore, TNF, IFN-gamma, and TGF-beta seem to be related to the full display of the suppressor function of MAC-induced M phi s. However, TNF-alpha and IFN-gamma but not TGF-beta were substantially lacking in inhibitory action against SPC mitogenesis, when added exogenously. Hence, it is unlikely that TNF-alpha and INF-gamma directly modulated the proliferative response of T cells. On the other hand, both TNF-alpha and IFN gamma potentiated the effector function of the suppressor M phi s. Because their suppressor activity was severely reduced by NG-monomethyl-L-arginine and aminoguanidine, nitric oxide (NO) synthase inhibitors, an NO-dependent mechanism is important for the expression of the immunosuppressive function of MAC-induced M phi s. Moreover, because these M phi s seem to produce a substantial amount of TNF-alpha in membrane-bound form, cell-to-cell contact might be needed for efficient expression of their suppressor action on target T cells. PMID- 7499971 TI - Macrophage apoptosis in the absence of active interleukin-1 beta-converting enzyme. AB - The interleukin-1 beta converting enzyme (ICE) is the cysteine proteinase responsible for cleaving the 31-kDa interleukin-1 beta (IL-1 beta) precursor to its active 17-kDa form. In lipopolysaccharide-stimulated cultured macrophages, induction of apoptosis but not necrosis effectively induces conversion of the IL 1 beta precursor to its mature form and results in the concomitant release of the mature cytokine from the cell. To determine whether ICE activity is required for macrophage apoptosis, we have exposed macrophages either to 5 mM ATP or to alloreactive cytolytic T lymphocytes (CTL) in the absence and presence of the ICE inhibitor peptide YVAD-chloromethylketone (YVAD-emk). Activated cells treated with YVAD-emk and ATP or CTL showed no mature IL-1 beta in either the cell lysates or the culture supernatants, indicating effective inhibition of ICE activity; however, the YVAD-treated macrophages showed no detectable change in 51Cr release or nuclear fragmentation, indicating failure to inhibit apoptotic cell death. Thus, in these cells, YVAD-emk uncouples IL-1 beta processing and apoptosis. PMID- 7499972 TI - Activation of inducible nitric oxide synthase gene in murine macrophages requires protein phosphatases 1 and 2A activities. AB - The purpose of these studies was to identify phosphatase activities required for the production of nitric oxide in murine macrophages exposed to lipopolysaccharide (LPS), synthetic lipopeptide (LPP), and mouse interferon-gamma (IFN-gamma). The in vitro treatment of macrophages with IFN-gamma and LPS or IFN gamma and LPP resulted in production of NO, which was inhibited by addition of the specific phosphatase 1 and 2A (PP1/2A) inhibitors okadaic acid (OA), calyculin A, and cantharidin (but not the nonactive analogues okadaic acid tetraacetate and 1,4-dimethylendothall). OA suppressed the accumulation of steady state inducible NO synthase (iNOS) mRNA and iNOS protein (without alteration of their stability). The cytosol and nuclei of control macrophages contained large amounts of PP1/2A activities that were inhibited by OA in a dose-dependent manner. Taken together, these data indicate that PP1/2A activities are involved in the regulation of iNOS gene expression in murine macrophages. PMID- 7499973 TI - Physical and chemical properties of a new root-end filling material. AB - This study determined the chemical composition, pH, and radiopacity of mineral trioxide aggregate (MTA), and also compared the setting time, compressive strength, and solubility of this material with those of amalgam, Super-EBA, and Intermediate Restorative Material (IRM). X-ray energy dispersive spectrometer in conjunction with the scanning electron microscope were used to determine the composition of MTA, and the pH value of MTA was assessed with a pH meter using a temperature-compensated electrode. The radiopacity of MTA was determined according to the method described by the International Organization for Standardization. The setting time and compressive strength of these materials were determined according to methods recommended by the British Standards Institution. The degree of solubility of the materials was assessed according to modified American Dental Association specifications. The results showed that the main molecules present in MTA are calcium and phosphorous ions. In addition, MTA has a pH of 10.2 initially, which rises to 12.5 three hours after mixing. MTA is more radiopaque than Super-EBA and IRM. Amalgam had the shortest setting time (4 min) and MTA the longest (2 h 45 min). At 24 h MTA had the lowest compressive strength (40 MPa) among the materials, but it increased after 21 days to 67 MPa. Finally, except for IRM, none of the materials tested showed any solubility under the conditions of this study. PMID- 7499974 TI - A comparison of three different root canal sealers when used to obturate a moisture-contaminated root canal system. AB - Sealing ability of three classes of endodontic sealers, in moisture-contaminated canals, was evaluated. A total of 120 single-rooted extracted teeth were divided into three groups. The groups were obturated with either Procosol, Sealapex, or Ketac-Endo using saline as a moisture contaminant in all cases. Half (20 teeth) of each group was placed in a saline storage medium for 9 months. The remainder of the samples were placed in India ink, under a vacuum, cleared, and the amount of dye penetration measured under magnification. The stored samples were similarly treated after the 9-month storage period. In a moisture-contaminated canal, Procosol exhibited the least amount of apical leakage, followed by Sealapex, then Ketac-Endo. There was no significant difference in the amount of dye penetration after 9 months of storage over that found initially, which indicates that the deleterious effects of moisture contamination occur during the initial placement and setting reaction. PMID- 7499975 TI - An ultrastructural study of debris retention by endodontic reamers. AB - The aim of this study was to determine the structure and the ultrastructure of the organo-mineral coating retained by endodontic reamers. Premolars extracted for orthodontic reasons were instrumented using Rispi Micromega reamers mounted on a counterangle Giromatic. One group of premolars was instrumented without irrigation, a second group was instrumented with irrigation using physiological saline, and a third group was instrumented with irrigation using Dakin. After instrumentation, the specimens were observed by scanning electron microscopy. Two microscopic techniques were used for each specimen. Secondary electron imaging was used to examine the amount of material adhering to the surfaces of the Rispi reamers after instrumentation, whereas backscattered electron imaging was used to determine the variation in the organo-mineral composition of the smear layer adhering to the reamers. The deeper layer adhering to the surface of the reamers was predominantly organic in nature no matter which instrumentation or irrigation technique was used. PMID- 7499976 TI - A three-dimensional finite-element stress analysis of an endodontically prepared maxillary central incisor. AB - This study is an application of a three-dimensional Finite-Element Method to investigate the changes in stress characteristics of a prepared maxillary central incisor. The purpose of this study was to analyze stress distributions in this tooth after simulated canal preparation and static loading. A maxillary central incisor was embedded in acrylic, sectioned, photographed, and digitized. A three dimensional finite-element model was generated by a computer and appropriately modified to simulate canal preparation. Data identified the highest stress magnitudes to be located between the middle and coronal thirds of the root; an area clinically observed to be prone to fracture during treatment. In addition, the magnitude of generated stresses was directly correlated with the simulated prepared canal diameter. The development of a validated three-dimensional finite element method could identify areas that may predispose a tooth to structural failure during condensation loads. PMID- 7499977 TI - Evaluation of root thickness in curved canals after flaring. AB - Root thickness in the mesial canals of lower first and second molars was compared before and after flaring by using a muffle system. Gates Glidden burs or M-Series Canal Openers were used with either a straight up-and-down motion or with an anticurvature motion. It was found that Gates Glidden burs used straight up and down removed statistically more dentin at the level of the furcation than Canal Openers used straight up and down or in an anticurvature fashion. At a level 2 mm apical to the furcation, Gates Glidden burs used in an anticurvature fashion removed statistically more dentin than any other method tested. PMID- 7499978 TI - Effect of traumatic occlusion on periapical lesions in rats. AB - The effect of traumatic occlusion on periapical lesions in rats was investigated histologically and histometrically. Rats were divided equally into groups A to D. Rats in group A received no treatment; in group B, rats received pulpal exposure of the left mandibular first molar; in group C, a resin plate was cemented onto the occlusal surface of the corresponding maxillary molar; and in group D, the molar pulp was exposed and the resin plate was installed. At 1 and 2 wk, compression of the periodontal ligament and inflammation were less in group D than in group B. Lesions in the periapical periodontal ligament at 1, 2, and 4 wk in group D were significantly smaller than those in group B. This study suggests that traumatic occlusion delayed the enlargement of the periapical lesions in rats. PMID- 7499979 TI - Microleakage of posts used to restore endodontically treated teeth. AB - The purpose of this study was to quantitate the microleakage of various post systems using a fluid filtration system. Eighty extracted human single-rooted teeth were used. The crowns were removed, the canals prepared, and the posts were placed. The samples were divided into three control groups and five experimental groups. The post systems tested were: (a) stainless steel posts cemented with zinc phosphate cement or (b) polycarboxylate cement; (c) a composite resin; (d) composite resin after use of a dentin bonding agent; and (e) composite resin after use of a dentin conditioner and dentin bonding agent. The results showed that none of the post systems tested were capable of consistently achieving a fluid-tight seal. PMID- 7499980 TI - Incidence and position of the canal isthmus. Part 1. Mesiobuccal root of the maxillary first molar. AB - The mesiobuccal roots of 50 randomly selected maxillary first molars were examined to assess the incidence and position of the canal isthmus. Transverse serial sections of the apical 6 mm of each root were prepared in 1-mm increments. The apical side of each section was stained with methylene blue dye, viewed with a surgical operating microscope, and videotaped. Forty percent of the roots had one canal, whereas 60% had two canals. None of the sections had more than two main canals. The incidence of an isthmus was highest in the apical 3- to 5-mm levels. In teeth that had two canals, the 4-mm sections contained a complete or partial isthmus 100% of the time. The concept of a partial isthmus was presented. Failure to deal with the isthmus may explain why some posterior teeth do not heal completely following endodontic surgery. PMID- 7499981 TI - Evaluation of success and failure after endodontic therapy using a glass ionomer cement sealer. AB - The purpose of this multicenter, prospective clinical study was to assess the treatment results following endodontic therapy using a glass ionomer cement sealer (Ketac-Endo) and to relate the results to various clinical factors. A total of 486 teeth were treated by three operators, using the "standardized technique" for canal preparation and either single cone or laterally condensed gutta-percha, in one or multiple treatment sessions. Six to 18 months postoperatively, the treatment results were assessed clinically and radiographically, and related to preoperative, intraoperative, and postoperative factors using a chi 2 analysis with 5% level of significance. Of 378 followed-up teeth, there was a 78.3% success, 15.6% incomplete healing, and 6.1% failure. Statistically, differences in the results were related to the number of canals (p < 0.04), primary treatment and retreatment (p < 0.02), pulp vitality (p < 0.001), periapical lesion (p < 0.001), preoperative symptoms (p < 0.003), operative complications (p < 0.001), and absence of restoration (p < 0.03). It was concluded that these treatment results were compatible with those reported in previous studies, and supported the clinical use of Ketac-Endo as an acceptable endodontic sealer. PMID- 7499983 TI - Solution for the isolation of the working field in a difficult case of root canal therapy. AB - A technique for the isolation of a posterior tooth with large distogingival caries and which needed endodontic therapy is described. To ensure a perfect seal, avoiding contamination or infiltration of the chemicals used for irrigation during the root canal therapy, a direct resin temporary restoration was done before placing the rubber dam as required for endodontic treatment. PMID- 7499982 TI - Evaluation of the effect of delayed management of traumatized permanent teeth. AB - This investigation studied the complication that could arise from delayed management of traumatized permanent teeth. Patients reported to the primary care clinic at the College of Dentistry, King Saud University, seeking treatment of traumatized anterior teeth were evaluated. Esthetic consideration (64 patients) and pain, swelling, or discomforts (50 patients) were the main reasons the patients came to the clinic. Clinical and radiographic examinations, as well as history of the trauma, were recorded. It was found that most of the patients had a treatment delay exceeding 1 month. Initial pulp exposure resulted in 100% pulp necrosis. When a fracture involved both enamel and dentin, the frequency of pulp necrosis was 53%. External root resorption and pulp calcification were seen in few teeth. It was concluded that the figures for pulp necrosis could be misleading, because there might have been a bias when only patients with problems sought treatment. In addition, a trauma awareness educational program should be developed to encourage parents and the public to seek immediate dental treatment. Furthermore, a proper diagnostic test for pulp and periodontal complication should be done in patients with initially nontreated dental injuries. PMID- 7499984 TI - Clinical audit: more research is required. PMID- 7499985 TI - Randomised controlled trials of physical activity promotion in free living populations: a review. AB - OBJECTIVES: To review evidence on the effectiveness of trials of physical activity promotion in healthy, free living adults. To identify the more effective intervention programmes. METHODS: Computerised databases and references were searched. Experts were contacted and asked for information about existing work. INCLUSION CRITERIA: Randomised controlled trials of healthy, free living adult subjects, where exercise behaviour was the dependent variable were included. CONCLUSIONS: Ten trials were identified. The small number of trials limits the strength of any conclusions and highlights the need for more research. No UK based studies were found. Previously sedentary adults can increase activity levels and sustain them. Promotion of these changes requires personal instruction, continued support, and exercise of moderate intensity which does not depend on attendance at a facility. The exercise should be easily included into an existing lifestyle and should be enjoyable. Walking is the exercise most likely to fulfil these criteria. PMID- 7499986 TI - Perinatal mortality in a first generation immigrant population and its relation to unemployment in The Netherlands. AB - STUDY OBJECTIVE: To consider the association between biological and social risk factors and perinatal mortality in an ethnically mixed population in Amsterdam, The Netherlands. DESIGN: This was a matched case-control study. Cases included all registered stillborn infants and all registered liveborn infants who died within seven days of birth. Controls were selected from infants remaining alive. Each case was matched with two controls by date of registration. SETTING: Civil registry of births and deaths, Amsterdam, The Netherlands, 1975-80. PATIENTS: All 666 babies who died in the perinatal period and 1332 controls selected from the liveborn survivors. OUTCOME: Perinatal mortality. MAIN RESULTS: Perinatal mortality was independently associated with the father's and mother's employment status, maternal age, parity, and infant sex, but not with the father's or mother's country of birth. CONCLUSIONS: Employment status and not country of birth should be the main focus in studies of perinatal mortality in this population of mixed ethnicity. Future studies on selected behavioural, socio economic, and cultural factors are needed to provide a better understanding of the causes of increased perinatal mortality among families in which the parents are unemployed. PMID- 7499987 TI - Socioeconomic factors and mortality in urban settings: the case of Barcelona, Spain. AB - STUDY OBJECTIVE: This study aimed to describe the relationship between health and socioeconomic indicators in the 38 neighbourhoods of the city of Barcelona, Spain. DESIGN: Mortality data for 1983-89 and socioeconomic data for each of the 38 neighbourhoods of Barcelona were used. Mortality indicators used were the comparative mortality figure, the ratio of potential years of life lost, and life expectancy at birth. Socioeconomic indicators were the percentage of unemployed, the percentage of illiteracy, monthly telephone usage, the average power and age of cars, and the average rateable value of buildings and of land. The statistical correlation between socioeconomic indicators and mortality indicators was studied by Spearman's rank correlation coefficient. SETTING: The 38 neighbourhoods of Barcelona, Spain. MEASUREMENTS AND MAIN RESULTS: The comparative mortality figure ranged from 87.41-152.43 and the ratio of potential years of life lost from 74.94 237.31 in both sexes. Both the absolute difference and the ratio of the value for the neighbourhood with lowest mortality and that with highest mortality were larger when premature mortality was examined. Life expectancy at birth ranged from 64.77-75.32 years in men and 75.04-81.51 in women. All correlations between mortality and socioeconomic indicators were high and statistically significant: the higher the unemployment and illiteracy levels and the older the cars, the greater the comparative mortality figure and ratio of potential years of life lost, and the lower the life expectancy (negative correlations). Conversely, the higher the telephone use, the more powerful the cars, and the greater the rateable value, the lower the mortality (negative correlations) and the greater the life expectancy. These correlations were greater in males than in females. The highest correlations were with illiteracy. CONCLUSIONS: This study has detected significant differences in mortality in a large town in the Mediterranean region of Europe. PMID- 7499988 TI - Factors associated with weight for height and skinfold thickness in British children. AB - STUDY OBJECTIVE: To examine the associations of social and biological factors with measures of obesity in children. DESIGN: The study had a cross sectional design. SETTING: The analyses were based on data from two national study of health and growth cross sectional surveys. The "representative sample" comprised 1990 data from 22 English areas and 1990-91 data from 14 Scottish areas; the "inner city sample" comprised 1991 data from 20 English areas. PARTICIPANTS: The subjects were primary school children aged mainly 5-11 years living in England and Scotland. The "representative" sample included 10,628 children--6463 living in England and 4165 living in Scotland. The "inner city" sample included 7049 children--2183 white, 1124 Afro-Caribbean, 2696 Indian subcontinent, and 1046 from other groups. Due to missing values on continuous variables, 8374 children were included in the analyses. MEASUREMENTS AND MAIN RESULTS: The relation between social environment and childhood overweight was studied using several indicators of obesity. Triceps, subscapular, the sum of triceps and subscapular skinfolds, and weight for height were used as dependent variables. The analyses were carried out in two stages. Firstly, multiple linear regression analyses were used to assess the factors associated with dependent variables treated as continuous. Secondly, multiple linear logistic regression analyses were used to examine the association between independent factors and overweight and fatness defined as binary variables. Birth weight, mother's body mass index (BMI), and father's BMI were consistently associated (p < 0.001) in all models and were the variables that contributed most to the explained variation in the dependent variables. In the multiple regression analyses there was a consistent interaction between the effects of ethnic origin and family size on each outcome variable. In the logistic regression analyses the interaction was not significant, and highly significant associations between both overweight and fatness with the number of children were shown. Ethnic group was not significantly associated with overweight but it was with fatness. The strengths of the remaining significant associations were slight and inconsistent in relation to the dependent variables or the type of analysis. CONCLUSION: Very few variables were associated with measures of overweight and fatness. The only useful factor that was highly associated with all measures of fatness was the parents' BMI. Strategies to prevent childhood obesity should be aimed at the total population and special emphasis should be placed on families in which one or both parents are overweight. PMID- 7499989 TI - Sickness absence in the Whitehall II study, London: the role of social support and material problems. AB - STUDY OBJECTIVE: To investigate the role of social supports, social networks, and chronic stressors: (i) as predictors of sickness absence; and (ii) as potential explanations for the socioeconomic gradient in sickness absence. DESIGN: A prospective cohort study (Whitehall II study) with sociodemographic factors, health and social support measured at baseline, and spells of sickness absence measured prospectively. SETTING: Twenty London based non-industrial departments of the British civil service. PARTICIPANTS: Participants were civil servants (n = 10,308), aged 35-55 years at baseline, of whom 67% (6895) were men and 33% (3413) were women. The overall response rate for Whitehall II was 73% (74% for men and 71% for women). The analysis is based on 41% of the sample who had data on reasons for sickness absence and were administered all social support questions. Only 4.3% of participants did not complete all necessary questions and were excluded. MEASUREMENTS AND MAIN RESULTS: High levels of confiding/emotional support from the "closest person" predicted higher levels of both short and long spells of sickness absence. After adjusting for baseline physical and psychological health the effects were increased, suggesting that high levels of confiding/emotional support may encourage illness behaviour rather than generate illness. Social network measures showed a consistent but less striking pattern. Increased levels of negative aspects of social support resulted in higher rates of sickness absence. Material problems strongly predicted sickness absence, but the effect was diminished once adjustment for the covariables was made, suggesting that health status may be functioning as an intervening variable between chronic stressors and sickness absence. In addition, social support may buffer the effects of chronic stressors. Social support did not contribute to explaining the gradient in sickness absence by employment grade beyond that explained by the baseline covariables. CONCLUSIONS: Sickness absence from work is a complex phenomenon, combining illness and coping behaviours. High levels of confiding/emotional support, although not entirely consistent across samples, may either encourage people to stay at home when they are ill or may be accompanied by more social obligations at home prolonging sickness absence. Negative aspects of close relationships may jeopardize health and hence increase sickness absence. PMID- 7499990 TI - Health behaviours explain part of the differences in self reported health associated with partner/marital status in The Netherlands. AB - STUDY OBJECTIVE: To describe the differences in health behaviours in disparate marital status groups and to estimate the extent to which these can explain differences in health associated with marital status. DESIGN: Baseline data of a prospective cohort study were used. Directly age standardised percentages of each marital group that engaged in each of the following behaviours--smoking, alcohol consumption, coffee consumption, breakfast, leisure exercise, and body mass index -were computed. Multiple logistic regression models were fitted to estimate the health differences associated with marital status with and without control for differences in health behaviours. SETTING: The population of the city of Eindhoven and surrounding municipalities (mixed urban-rural area) in The Netherlands in March 1991. PARTICIPANTS: There were 16,311 men and women, aged 25 74 years, and of Dutch nationality. MAIN RESULTS: There were differences in relation to marital status for each health behaviour. Married people were more likely to practise positive health behaviours (such as exercise and eating breakfast) and less likely to engage in negative ones (such as smoking or drinking heavily) than the other groups. Control for all six health behaviours could explain an average of 20-36% of the differences in perceived and general health and subjective health complaints. CONCLUSIONS: Differences in health behaviours explained a considerable amount, but not all, of the health differences related to marital status. Longitudinal data are necessary to confirm these findings; to determine whether the differences in health behaviours related to marital status are caused by selection effects or social causation effects; and to learn how social control, social support, and stress inter-relate to reinforce negative or to maintain positive health behaviours. PMID- 7499991 TI - Seasonal ataxia in western Nigeria: evaluation of the impact of health education on hospital prevalence. AB - STUDY OBJECTIVE: The study aimed to evaluate the impact of focused health education on the hospital prevalence of a seasonal ataxic syndrome which occurs exclusively in people of low income after eating the larvae of Anaphe venata Butler (Lepidoptera, Notodontidae). DESIGN: Retrospective impact evaluation study. SETTING: Wesley Guild Hospital, Ilesha, western Nigeria from where the syndrome was first described. METHODS: Hospital workers were informed of recent research findings on the aetiological role of Anaphe venata larvae consumption in the annual epidemics of seasonal ataxia. They were encouraged to educate hospital patients and their relatives and to ask them to pass this information to neighbours and other family members who are usually of the same social strata. The number of admissions for seasonal ataxia in the pre-intervention years (1990 and 1991) and in the intervention years (1992 and 1993) were extracted from records. The total number of casualty admissions between July and November of each year were also obtained. The period prevalence rates of the disease in each year of the study period were calculated and compared. RESULTS: The pre intervention prevalence rates were 1.27% and 1.59% for 1990 and 1991 respectively. The prevalence rates in the intervention years of 1992 and 1993 were 0.58% and 0.006%, representing a 65.5% reduction between 1991 and 1992, and a 99% reduction between 1992 and 1993. CONCLUSION: The health education technique used is an inexpensive but effective way of providing the intervention needed for the control of nutrition related diseases in the poor, largely illiterate segment of communities that are most vulnerable. PMID- 7499992 TI - Low back pain in Hong Kong: prevalence and characteristics compared with Britain. AB - STUDY OBJECTIVE: To compare the prevalence of low back pain and associated disability in Hong Kong with that in Britain, and to explore whether differences could be explained by certain known risk factors. DESIGN: A cross sectional survey with information collected at interview. Findings were compared with those from an earlier survey in Britain. SETTING: Two housing blocks in Hong Kong. SUBJECTS: Altogether 288 men and 364 women aged 18 years or older, who were resident in the two housing blocks and agreed to interview (response rate = 80%). MAIN RESULTS: Thirty nine per cent (95% confidence interval (CI) 34%, 44%) of interviewees reported having had low back pain at some time, and 21% (95% CI 18%, 25%) had had low back pain in the past 12 months. After standardisation for age and sex, all of the back symptoms examined were substantially less common in Hong Kong than in Britain. The one year period prevalence of low back pain was associated with occupational lifting (in both sexes) and with tall stature (in men only). Subjects tended to carry out less heavy lifting at work and to be shorter than participants in the earlier British study but these differences did not completely explain their lower prevalence of back pain. CONCLUSIONS: The findings indicate a lower prevalence of back symptoms in Hong Kong than Britain that is partly explained by differences in stature and occupational lifting. In addition, Hong Kong people may have a higher threshold for reporting symptoms, or they may differ in their exposure to other, unrecognised risk factors. PMID- 7499994 TI - Coeur en sante St-Henri--a heart health promotion programme in a low income, low education neighbourhood in Montreal, Canada: theoretical model and early field experience. AB - STUDY OBJECTIVE: Coeur en sante St-Henri is a five year, community based, multifactorial, heart health promotion programme in a low income, low education neighbourhood in Montreal, Canada. The objectives of this programme are to improve heart-healthy behaviours among adults of St-Henri. This paper describes the theoretical model underlying programme development as well as our early field experience implementing interventions. DESIGN: The design of the intervention programme is based on a behaviour change model adapted from social learning theory, the reasoned action model, and the precede-proceed model. The Ottawa charter for health promotion provided the framework for the development of specific interventions. Each intervention is submitted to formative, implementation, and impact evaluations using simple and inexpensive methods. PARTICIPANTS: The target population consists of adults living in St-Henri, a neighbourhood of 23,360 residents. Because of costs constraints, the intervention strategy targets women more specifically. The community is one of the poorest in Canada with 46% of the population living below the poverty line and 20% being very poor. The age-sex adjusted ischaemic heart disease mortality in 1985-87 was 317 per 100,000 compared with 126 per 100,000 in an affluent adjacent neighbourhood. RESULTS: Thirty nine distinct interventions have been developed and tested in the community, eight related to tobacco, 10 to diet, seven to physical activity, and 14 which are multifactorial. The interventions include smoking cessation and healthy recipes contests, a menu labelling and healthy food discount programme in restaurants, a point of choice nutrition education campaign, healthy eating and smoking cessation workshops, a walking club, educational material, print and electronic media campaigns, heart health fairs, and community events. CONCLUSION: An integrated heart health promotion programme is feasible in low income urban neighbourhoods but not all interventions are successful. Such a programme requires substantial energy and resources as well as long term commitment from public health departments. PMID- 7499993 TI - Coeur en sante St-Henri--a heart health promotion programme in Montreal, Canada: design and methods for evaluation. AB - STUDY OBJECTIVE: This paper describes the objectives, design, and methods of evaluation of the impact of the coeur en sante St-Henri programme, as well as selected results from the evaluation to date. It discusses the possible effects of study design choices made to maintain the impact evaluation within budget. DESIGN: The impact of the programme is evaluated in a community trial which compares the prevalence of cardiovascular disease behavioural risk factors before and after programme implementation in the intervention and a matched comparison community, in both longitudinal cohort and independent sample surveys. In addition, repeated independent sample surveys are conducted in the intervention community to monitor awareness of and participation in the programme. PARTICIPANTS: The baseline sample for both the longitudinal cohort and independent sample surveys included 849 subjects from the intervention community (79.3% of 1071 eligible subjects--8.0% could not be contacted and 12.6% refused) and 825 subjects from the comparison community (77.8% of 1066 eligible subjects- 6.6% could not be contacted and 15.6% refused). The two surveys on awareness and participation conducted to date, included 461 (71.0% of 649 eligible subjects) and 387 (67.9% of 570 eligible subjects) subjects respectively from the intervention community. MEASUREMENTS: Baseline data for the longitudinal cohort and independent sample surveys on behavioural risk factor outcomes including use of tobacco, physical activity behaviour, high fat diet, and behaviours related to blood pressure and cholesterol control were collected in 35 minute telephone interviews in both the intervention and comparison communities. Data on awareness of and participation in the programme were collected in 10 minute interviews in the intervention community only in two independent sample surveys conducted seven and 22 months respectively after the baseline survey. RESULTS: With the exception of smoking, the intervention and comparison communities were similar at baseline with regard to the prevalence of behavioural risk factors studied. Awareness of the coeur en sante programme increased from 64.1% in January 1993 to 72.9% 15 months later. Participation in the programme increased from 21.3% to 33.7%. CONCLUSIONS: This paper presents background information on the evaluation of the impact of the coeur en sante programme, as a reference for future publications. PMID- 7499996 TI - A measurement of social support in epidemiological research: the social experiences checklist tested in a general population in The Netherlands. AB - STUDY OBJECTIVE: This study aimed to examine in a general population the psychometric qualities of an instrument designed to measure positive and negative social experiences that had been developed in a clinical setting. DESIGN: The Netherlands monitoring project on cardiovascular disease risk factors, a large scale population based study (comprising 36,588 men and women aged 20 to 59 years) carried out in three Dutch towns (Amsterdam, Doetinchem, and Maastricht) offered the possibility of testing the strength of this instrument cross sectionally. MEASUREMENTS AND MAIN RESULTS: The social experiences checklist (SEC) which resulted from a research project on the quality of life of cancer patients was used. The independence of positive and negative experiences was confirmed. The reliability of both the positive and negative experiences dimension was good (Cronbach's alpha = 0.82 and 0.72 respectively). In accordance with the results of a study on cancer patients, the theoretically derived four dimensions in the experience of social support did not seem to be independent. The validity of the SEC was confirmed by Pearson correlations with neuroticism and coping styles. Neuroticism seemed to be negatively correlated with positive social experiences and was positively correlated with negative social experiences. The coping style of seeking information and direct action was positively correlated with positive social experiences. Coping by withdrawal was negatively correlated with negative social experiences. Women and highly educated people seemed to have more positive and fewer negative social experiences than men and people with less education. Younger people had more positive social experiences than older people. The oldest group in the study, those aged 50 to 59, reported fewer negative social experiences than any other age group. CONCLUSIONS: Similar results were found in a study of cancer patients. This underlines the usefulness of the instrument not only for cancer patients but also in survey research in a general population. PMID- 7499995 TI - Measuring quality of life in patients with myocardial infarction or stroke: a feasibility study of four questionnaires in The Netherlands. AB - OBJECTIVE: To test in patients with a history of myocardial infarction or stroke the feasibility of four quality of life measurements--the Nottingham health profile (NHP), the heart patients psychological questionnaire (HPPQ), the sickness impact profile (SIP), and the hospital anxiety and depression scale (HAD). DESIGN: Subjects were tested and retested after an interval of 14 days: questionnaires were self assessed. SUBJECTS: Participants were randomly selected from the Rotterdam stroke data bank (stroke patients; n = 16, mean (SD) age 66.0 (11.0) years and from the population based Rotterdam study (myocardial infarction; n = 20, mean (SD) age 72.7 (7.9) years, controls; n = 17, mean (SD) age 72.8 (7.3) years. MEASUREMENTS AND MAIN RESULTS: Mean (SD) administration times for the NHP, HPPQ, SIP, and HAD were 7.9 (3.5), 10.5 (4.3), 21.0 (9.8), and 5.5 (2.8) minutes respectively. On average, the test-retest reliability was good, with Spearman correlations ranging from 0.31 to 0.95. In spite of the limited size of the study, all instruments were able to show differences between the study groups. For instance, median SIP total scores for myocardial infarction and stroke patients were 12.4 (interquartile range 7.0-19.1) and 11.4 (5.9-15.4) respectively, compared with 7.7 (3.7-11.3) in the control group (p values of 0.04 and 0.14 respectively). CONCLUSIONS: This study suggests that use of the four instruments tested may be feasible and reliable for assessing aspects of quality of life in patients with a history of a myocardial infarction or stroke. PMID- 7499997 TI - A method for estimating baseline health care costs. AB - STUDY OBJECTIVES: Studies estimating the cost of specific illnesses do not generally take into account the fact that health care costs would have been incurred in the absence of the disease of interest. The goal of this study was to develop a method of estimating age specific baseline health care costs. These costs were calculated for Australian men, and their magnitude was compared with the costs of caring for men with HIV infection. DESIGN: Information about health service usage was obtained from the 1989-90 national health survey and linked with data on the costs of services to obtain average monthly costs for individual and total health services. SETTING: The Australian community. PARTICIPANTS: Average total health service costs per man per month were $103 (Australian). Hospital admissions comprised approximately 40% of these costs and casualty/outpatient visits, consultations with a doctor, and prescribed medication comprised 10%, 13%, and 12%, respectively. Costs increased with age, from around $60 per month for men aged 20-39 years to $213 per month for men aged 60 and over. CONCLUSION: Baseline costs comprised around 18% of health care costs for men with asymptomatic HIV infection, but less than 1% of costs for men with AIDS. These estimates provide an essential baseline for determining the costs attributable to specific diseases. PMID- 7499998 TI - General practice consultation in childhood in The Netherlands: sociodemographic variation. PMID- 7500000 TI - Smoking and health promotion in Nazi Germany. PMID- 7499999 TI - Short form 36 questionnaire. PMID- 7500001 TI - Recovery of function, peripheral sensitization and sensory neurone activation by novel pathways following axonal injury in Aplysia californica. AB - Recovery of behavioural and sensory function was examined following unilateral pedal nerve crush in Aplysia californica. Nerve crush that transected all axons connecting the tail to the central nervous system (CNS) eliminated the ipsilateral tail-evoked siphon reflex, whose sensory input travels in the crushed tail nerve (p9). The first reliable signs of recovery of this reflex were observed within 1 week, and most animals displayed tail-evoked siphon responses within 2 weeks. Wide-dynamic-range mechanosensory neurons with somata in the ventrocaudal (VC) cluster of the ipsilateral pleural ganglion exhibited a few receptive fields (RFs) on the tail 3 weeks after unilateral pedal nerve crush, indicating that the RFs had either regenerated or been reconnected to the central somata. These RFs were smaller and sensitized compared with corresponding RFs on the contralateral, uncrushed side. Centrally conducted axon responses of VC sensory neurones to electrical stimulation distal to the nerve crush site did not reappear until at least 10 days after the crush. Because the crush site was much closer to the CNS than to the tail, the failure of axon responses to be restored earlier than the behavioural responses indicates that early stages of reflex recovery are not due to regeneration of VC sensory neurone axons into the tail. Following nerve crush, VC sensory neurones often could be activated by stimulating central connectives or peripheral nerves that do not normally contain the sensory neurone's axons. These results suggest that recovery of behavioral function after nerve injury involves complex mechanisms, including regenerative growth of axotomized VC sensory neurones, sensitization of regenerating RFs and sprouting of VC sensory neurone fibres within the CNS. Furthermore, the rapidity of behavioural recovery indicates that its initial phases are mediated by additional mechanisms, perhaps centripetal regeneration of unidentified sensory neurones having peripheral somata, or transient reconnection of proximal and distal stumps of axotomized VC cells. PMID- 7500002 TI - Peripheral regeneration and central sprouting of sensory neurone axons in Aplysia californica following nerve injury. AB - Morphological methods were used to examine injury-induced growth of peripheral and central axons of nociceptive mechanosensory neurones in the ventrocaudal (VC) clusters of the pleural ganglia of Aplysia californica. Pedal nerve crush transected all axons in the nerve while leaving the overlying sheath largely intact. Immunohistochemical staining was performed with an antibody to a sensory neurone-specific peptide, sensorin-A. Following bilateral crush of pedal nerve p9, which innervates the tail, sensorin-A immunofluorescence was lost distal to the crush site within 2 days. Fine immunopositive fibres began to invade the crush region within 5 days. These fibres arborized in the crush region and gradually extended down the crushed nerve. Immunopositive fibres were found near the tail within 3 weeks. Similar results were obtained after injecting individual sensory neurone somata in the tail/p9 region of the VC cluster with biocytin. Biocytin injections and horseradish peroxidase injections 3 weeks after ipsilateral pedal nerve crush revealed new fibres projecting rostrally from the tail/p9 region of the VC cluster and entering the pleural-cerebral and pleural abdominal connectives. Such projections were never observed in control, uncrushed preparations. These results demonstrate that nerve injury triggers extensive growth of both peripheral and central processes of the VC sensory neurones. PMID- 7500003 TI - The structure and physical properties of invertebrate and primitive vertebrate arteries. AB - Light and electron microscopy and in vitro inflation experiments were conducted on the aortae of three different invertebrate species: the lobster Homarus americanus, the horseshoe crab Limulus polyphemus and the whelk Busycon contrarium. Inflation experiments were also performed on the aortae of two species of primitive vertebrates, the sea lamprey Petromyzon marinus and the Atlantic hagfish Myxine glutinosa. The inflation experiments demonstrated similar overall biomechanical properties in each case, despite the existence of differences in tissue structure. The vessels were compliant at low strains, but demonstrated nonlinear elasticity, increasing in stiffness as strains increased; this property could act as protection against artery wall rupture. The vessels of the lamprey, hagfish and lobster are capable of acting as fairly efficient elastic reservoirs and of smoothing blood flow during circulation as they had low hysteresis values (13-18%). The aortae of the horseshoe crab and whelk, if performing this function, have much higher energy losses, up to more than 30% per cycle. The microscopy studies of the aortae of the lobster, horseshoe crab and whelk revealed tissue structures which differ widely from each other as well as from the structures of the lamprey and hagfish. None of these arteries contained elastin, but all contained fibrillar material which differed in appearance, size and arrangement between species. These materials were conjectured to contribute to the elastic properties of the tissue. PMID- 7500005 TI - Influence of muscle length on work from trabecular muscle of frog atrium and ventricle. AB - The work capacity of segments of atrial and ventricular muscle from the frog Rana pipiens was measured as a function of muscle length using the work loop technique. Both the work done during shortening and the work required to re lengthen the muscle after shortening increased with muscle length. Net work increased with length up to a maximum, beyond which work declined. The optimum sarcomere length for work output was 2.5-2.6 microns for both atrial and ventricular muscle. Isometric force increased with muscle length to lengths well beyond the optimum for work output. Thus, the decline in work at long lengths is not simply a consequence of a reduction in the capacity of heart muscle to generate force. It is proposed that it is the non-linear increase in work required to re-lengthen muscle with increasing muscle length which limits net work output and leads to a maximum in the relationship between net work and muscle length. Extension of the results from muscle strips to intact hearts suggests that the work required to fill the ventricle exceeds that available from atrial muscle at all but rather short ventricular muscle lengths. PMID- 7500004 TI - Sodium-dependent norepinephrine-induced currents in norepinephrine-transporter transfected HEK-293 cells blocked by cocaine and antidepressants. AB - Transport of norepinephrine (NE+) by cocaine- and antidepressant-sensitive transporters in presynaptic terminals is predicted to involve the cotransport of Na+ and Cl-, resulting in a net movement of charge per transport cycle. To explore the relationship between catecholamine transport and ion permeation through the NE transporter, we established a human norepinephrine transporter (hNET) cell line suitable for biochemical analysis and patch-clamp recording. Stable transfection of hNET cDNA into HEK-293 (human embryonic kidney) cells results in lines exhibiting (1) a high number of transporter copies per cell (10(6)), as detected by radioligand binding and hNET-specific antibodies, (2) high-affinity, Na(+)-dependent transport of NE, and (3) inhibitor sensitivities similar to those of native membranes. Whole-cell voltage-clamp of hNET-293 cells reveals NE-induced, Na(+)-dependent currents blocked by antidepressants and cocaine that are absent in parental cells. In addition to NE-dependent currents, transfected cells posses an NE-independent mode of charge movement mediated by hNET. hNET antagonists without effect in non-transfected cells abolish both NE dependent and NE-independent modes of charge movement in transfected cells. The magnitude of NE-dependent currents in these cells exceeds the expectations of simple carrier models using previous estimates of transport rates. To explain our observations, we propose that hNETs function as ion-gated ligand channels with an indefinite stoichiometry relating ion flux to NE transport. In this view, external Na+ and NE bind to the transporter with finite affinities in a cooperative fashion. However, coupled transport may not predict the magnitude or the kinetics of the total current through the transporter. We propose instead that Na+ gates NE transport and also the parallel inward flux of an indeterminate number of ions through a channel-like pore. PMID- 7500006 TI - Interactions of cortisol and nitrogen metabolism in the ureogenic gulf toadfish Opsanus beta. AB - We examined the relationship between plasma cortisol levels, hepatic glutamine synthetase (GNS) activity and the form of nitrogen excreted (e.g. urea, ammonia) in undisturbed versus confined/crowded (acutely stressed) gulf toadfish. Specifically, we tested the hypothesis that acute increases in plasma cortisol levels are required to trigger the increase in GNS activity induced by the confinement/crowding stress. Toadfish responded to the stress of confinement/crowding with an initial cortisol surge (approximately 37 ng ml-1 at 2 h), which was rapidly cleared and was indistinguishable from resting levels (approximately 10 ng ml-1 by 24 h). Treatment of fish with metyrapone (which inhibits cortisol synthesis) successfully blocked the acute 2 h confinement/crowding-induced surge in plasma cortisol levels. Additionally, GNS activity in confined fish 24 h after metyrapone injection also did not differ from that of control fish, indicating that acute GNS activation probably requires the earlier (2 h) cortisol peak. In post-absorptive fish, a strong relationship between total nitrogen excretion rate and plasma cortisol levels was evident. The percentage of nitrogen excreted as ammonia was inversely related to liver GNS activity. However, GNS activity explained only part (at most 57%) of the variability in the percentage of nitrogen excreted as ammonia/urea, suggesting that this is not the sole factor setting the degree of ureogenesis. When toadfish are fed, the relationships between total nitrogen excretion rate and cortisol levels, and between percentage nitrogen excreted as ammonia and GNS activity, are virtually absent. Taken together, our results indicate that a stress response may be only one of several mechanisms by which ureogenesis is activated in gulf toadfish. PMID- 7500007 TI - DNA-dependent protein kinase is one of a subset of autoantigens specifically cleaved early during apoptosis. AB - Proteolytic cleavage of key substrates appears to be an important biochemical mechanism underlying the apoptotic process, and the centrality of interleukin 1 beta-converting enzyme (ICE)-like proteases as mediators of apoptosis has been suggested. The identification of the relevant substrates of the ICE protease family during apoptosis therefore constitutes a major challenge. Using human autoantibodies, we demonstrate here that a subset of autoantigens is specifically cleaved early during apoptosis. One of these cleaved molecules is identified as the catalytic subunit of the DNA-dependent protein kinase. The time courses of all proteolytic cleavages are identical and coincide with the onset of morphologic apoptosis. Furthermore, all cleavages share the same inhibition characteristics, which implicate an ICE-like activity(ies). We propose that cleavage of these autoantigens targets these molecules for an autoimmune response by revealing immunocryptic fragments in a proimmune apoptotic setting. Study of the immunogenicity of these fragments may yield insights into the autoimmune targeting of molecules. Moreover, the autoantibodies described will be valuable tools for the elucidation of mechanistically important proteolytic steps along the apoptotic pathway. PMID- 7500009 TI - Leukemia treatment in severe combined immunodeficiency mice by antisense oligodeoxynucleotides targeting cooperating oncogenes. AB - Transformation of hematopoietic cells by the p210bcr/abl tyrosine kinase appears to require the expression of a functional MYC protein, suggesting that simultaneous targeting of BCR-ABL and c-myc might be a rational strategy for attempting treatment of Phil-adelphia leukemia. To test this hypothesis, severe combined immunodeficiency mice injected with Philadelphia leukemic cells were treated systemically with equal doses of bcr-abl or c-myc antisense oligodeoxynucleotides (ODNs) or with both ODNs in combination. Compared with the mice treated with individual agents, the disease process was much slower in the group treated with both ODNs, as revealed by flow cytometry, clonogenic assay, and reverse transcriptase-polymerase chain reaction analysis to detect leukemic cells in mouse tissue cell suspensions, and by enumeration of liver metastases. The retardation of the disease process was positively correlated with a markedly increased survival of leukemic mice treated with both ODNs. These data demonstrate the therapeutic potential of targeting multiple cooperating oncogenes. PMID- 7500008 TI - In situ studies of the primary immune response to (4-hydroxy-3 nitrophenyl)acetyl. IV. Affinity-dependent, antigen-driven B cell apoptosis in germinal centers as a mechanism for maintaining self-tolerance. AB - Germinal centers (GCs) are the sites of antigen-driven V(D)J gene hypermutation and selection necessary for the generation of high affinity memory B lymphocytes. Despite the antigen dependence of this reaction, injection of soluble antigen during an established primary immune response induces massive apoptotic death in GC B cells, but not in clonally related populations of nonfollicular B lymphoblasts and plasmacytes. Cell death in GCs occurs predominantly among light zone centrocytes, is antigen specific, and peaks within 4-8 h after injection. Antigen-induced programmed death does not involve cellular interactions mediated by CD40 ligand (CD40L) or Fas; disruption of GCs by antibody specific for CD40L was not driven by apoptosis and C57BL/6.lpr mice, though unable to express the Fas death trigger, remained fully susceptible to soluble antigen. Single injections of antigen did not significantly decrease GC numbers or average size, but repeated injections during an 18-h period resulted in fewer and substantially smaller GCs. As cell loss appeared most extensive in the light zone, decreased GC cellularity after prolonged exposure to soluble antigen implies that the Ig- centroblasts of the dark zone may require replenishment from light zone cells that have survived antigenic selection. GC cell death is avidity-dependent; oligovalent antigen induced relatively little apoptosis and GC B cells that survived long exposures to multivalent antigen expressed atypical VDJ rearrangements unlikely to encode high affinity antibody. Antigen-induced apoptotic death in GCs may represent a mechanism for the peripheral deletion of autoreactive B cell mutants much as the combinatorial repertoire of immature B lymphocytes is censored in the bone marrow. PMID- 7500010 TI - CD30-mediated signaling promotes the development of human T helper type 2-like T cells. AB - We have recently shown that CD30, a member of the tumor necrosis factor/nerve growth factor receptor superfamily, is preferentially expressed by human T cell clones producing T helper (Th) type 2 cytokines. We report here that costimulation with an agonistic anti-CD30 monoclonal antibody enhanced antigen (Ag)-induced proliferation and cytokine secretion by established human Th2 and Th0 clones. Moreover, costimulation of peripheral blood mononuclear cells with the same anti-CD30 monoclonal antibody resulted in the preferential development of Ag-specific T cell lines and clones showing a Th2-like profile of cytokine secretion. In contrast, early blockade in bulk culture of CD30 ligand-CD30 interaction shifted the development of Ag-specific T cells towards the opposite (Th1-like) phenotype. Taken together, these data suggest that CD30 triggering of activated Th cells by CD30 ligand-expressing Ag-presenting cells may represent an important costimulatory signaling for the development of Th2-type responses. PMID- 7500011 TI - Inactivated influenza virus, when presented on dendritic cells, elicits human CD8+ cytolytic T cell responses. AB - Inactivated or subunit virus preparations have been excellent vaccines for inducing antibody responses. Generation of cytolytic T cell responses, however, is thought to require replicating virus, primarily to provide sufficiently large amounts of cytoplasmic proteins for processing and presentation on major histocompatibility complex class I molecules by antigen-presenting cells. Potent human CD8+ cytolytic T cell responses to live replicating influenza A virus are generated when dendritic cells are used as the antigen-presenting cells. Here, we demonstrate that dendritic cells pulsed with poorly replicating, heat- or ultraviolet-inactivated influenza virus, induce equally strong CD8+ cytolytic T lymphocyte responses. The cytotoxic T lymphocytes are generated in the apparent absence of CD4+ helper cells or exogenous cytokines. Active viral protein synthesis is not required to charge class I molecules on dendritic cells. When pulsed with inactivated virus, < 1% of dendritic cells express nonstructural protein 1, which is only synthesized in the infectious cycle. To be optimally effective, however, the inactivated virus must retain its fusogenic activity, and presumably access the cytoplasm of dendritic cells. The data indicate, therefore, that dendritic cells require only small amounts of viral protein to charge class I molecules, most likely via traditional class I processing pathways. These results reopen the potential use of inactivated virus preparations as immunogens for cytotoxic T lymphocyte responses. PMID- 7500012 TI - Molecules from Staphylococcus aureus that bind CD14 and stimulate innate immune responses. AB - Mammals mount a rapid inflammatory response to gram-negative bacteria by recognizing lipopolysaccharide (LPS, endotoxin). LPS binds to CD14, and the resulting LPS-CD14 complex induces synthesis of cytokines and up-regulation of adhesion molecules in a variety of cell types. Gram-positive bacteria provoke a very similar inflammatory response, but the molecules that provoke innate responses to these bacteria have not been defined. Here we show that protein free, phenol extracts of Staphylococcus aureus contain a minor component that stimulates adhesion of neutrophils and cytokine production in monocytes and in the astrocytoma cell line, U373. Responses to this component do not absolutely require CD14, but addition of soluble CD14 enhances sensitivity of U373 cells by up to 100-fold, and blocking CD14 on monocytes decreases sensitivity nearly 1,000 fold. Deletion of residues 57-64 of CD14, which are required for responses to LPS, also eliminates CD14-dependent responses to S. aureus molecules. The stimulatory component of S. aureus binds CD14 and blocks binding of radioactive LPS. Unlike LPS, the activity of S. aureus molecules was neither enhanced by LPS binding protein nor inhibited by bactericidal/permeability increasing protein. The active factor in extracts of S. aureus is also structurally and functionally distinct from the abundant species known as lipoteichoic acid (LTA). Cell stimulating activity fractionates differently from LTA on a reverse-phase column, pure LTA fails to stimulate cells, and LTA antagonizes the action of LPS in assays of IL-6 production. These studies suggest that mammals may use CD14 in innate responses to both gram-negative and gram-positive bacteria, and that gram positive bacteria may contain an apparently unique, CD14-binding species that initiates cellular responses. PMID- 7500013 TI - A hypoxia-responsive element mediates a novel pathway of activation of the inducible nitric oxide synthase promoter. AB - Picolinic acid, a catabolite of L-tryptophan, activates the transcription of the inducible nitric oxide synthase gene (iNOS) in IFN-gamma-treated murine macrophages. We performed functional studies on the 5' flanking region of the iNOS gene linked to a CAT reporter gene to identify the cis-acting element(s) responsible for the activation of iNOS transcription by picolinic acid. Transient transfection assays showed that the full-length iNOS promoter in the murine macrophage cell line ANA-1 was activated by the synergistic interaction between IFN-gamma and picolinic acid. Deletion or mutation of the iNOS promoter region from -227 to -209, containing a sequence homology to a hypoxia-responsive enhancer (iNOS-HRE), decreased picolinic acid- but not LPS-induced CAT activity by more than 70%. Functional studies using a tk promoter-CAT reporter gene plasmid demonstrated that the iNOS-HRE was sufficient to confer inducibility by picolinic acid but not by IFN-gamma or LPS. Electrophoretic mobility shift assays confirmed that picolinic acid alone induced a specific binding activity to the iNOS-HRE. Furthermore, we found that the iNOS-HRE activity was inducible by hypoxia and that hypoxia in combination with IFN-gamma activated the iNOS promoter in transient transfection assays and induced iNOS transcription and mRNA expression. These data establish that the iNOS-HRE is a novel regulatory element of the iNOS promoter activity in murine macrophages and provide the first evidence that iNOS is a hypoxia-inducible gene. PMID- 7500014 TI - The PapG-adhesin at the tip of P-fimbriae provides Escherichia coli with a competitive edge in experimental bladder infections of cynomolgus monkeys. AB - Human urinary tract infection is an infectious disease that depends on a series of host-microbial interactions. The bacteria first colonize the colon and then the periurethral/vaginal areas; they ascend to and infect first the bladder and then the kidneys. Expression of Escherichia coli P-fimbriae constitutes the strongest correlation to renal pathogenicity, but is also related to first-time cystitis in children. The role of P-fimbriae in the preceding steps in the infectious process is unknown. To examine this, we constructed, from a P fimbriated E. coli strain with a class II G-adhesin preferentially binding to globoside, one isogenic mutant lacking the G-adhesin and another isogenic mutant in which we replaced the papG class II allele with a class III adhesin preferentially binding to the Forssman antigen. We report here the comparison of the adhesin knockout mutant (DS17-8) and the class-switch mutant (DS17-1) with the wild-type (DS17) for in vivo colonization of the gut, vagina, and bladder of cynomolgus monkeys. It was recently shown that the class II tip G-adhesin is a prerequisite for acute pyelonephritis to occur in the monkey model in the absence of other kidney-specific adhesins or obstruction of the urinary flow. Here we show that it is not required for bladder infection but gives a competitive advantage in mixed infections. In the vagina and colon, the G-adhesin gives no competitive advantage. PMID- 7500016 TI - A transforming growth factor beta-like immunosuppressive factor in immunoglobulin G-binding factor. AB - Immunoglobulin G-binding factors (IgG-BF), which are produced by cells of the immune system, inhibit antibody production. In this paper, we show that transforming growth factor-beta (TGF-beta) suppresses secondary in vitro anti sheep red blood cell responses of mouse splenocytes and lipopolysaccharide- or anti-IgM-stimulated mouse B cell responses in a way similar to, and with the same kinetics as, rodent IgG-BF. Moreover, the immunosuppressive activity of IgG-BF was totally neutralized by polyclonal and monoclonal anti-TGF-beta antibodies and it eluted with TGF-beta by gel exclusion chromatography, suggesting that a TGF beta-like immunosuppressive factor is present in IgG-BF. We also show that TGF beta behaves as an IgG-BF since it binds to insolubilized IgG, but not to insolubilized F(ab')2 or bovine serum albumin. Altogether, the data support the concept of a biological role for TGF-beta in the IgG-mediated negative feedback of antibody responses. PMID- 7500015 TI - T cell receptor repertoire for a viral epitope in humans is diversified by tolerance to a background major histocompatibility complex antigen. AB - Two unusual characteristics of the memory response to the immunodominant Epstein Barr virus (EBV) epitope FLRGRAYGL, which associates with HLA B8, have provided an unique opportunity to investigate self tolerance and T cell receptor (TCR) plasticity in humans. First, the response is exceptionally restricted, dominated by cytotoxic T lymphocytes (CTL) with identical TCR protein sequences (Argaet, V. P., C. W. Schmidt, S. R. Burrows, S. L. Silins, M. G. Kurilla, D. L. Doolan, A. Suhrbier, D. J. Moss, E. Kieff, T. B. Sculley, and I. S. Misko. 1994. J. Exp. Med. 180:2335-2340). Second, CTL expressing this receptor are cross-reactive with the alloantigen HLA B* 4402 on uninfected cells (Burrows, S. R., R. Khanna, J. M. Burrows, and D. J. Moss. 1994. J. Exp. Med. 179:1155-1161). No CTL using this conserved public TCR could be reactivated from the peripheral blood of EBV exposed individuals expressing both HLA B8 and B*4402, demonstrating the clonal inactivation of potentially self-reactive T cells in humans. A significant FLRGRAYGL-specific response was still apparent, however, and TCR sequence analysis of multiple CTL clones revealed an oligoclonal TCR repertoire for this determinant within these individuals, using diverse V and J gene segments and CDR3 regions. In addition, a significant public TCR component was identified in which several distinct alpha/beta rearrangements are shared by CTL clones from a number of unrelated HLA B8+, B*4402+ donors. The striking dominance of public TCR in the response to this EBV epitope suggests a strong genetic bias in TCR gene recombination. Fine specificity analysis using peptide analogues showed that, of six different antigen receptors for FLRGRAYGL/HLA B8, none associate closely with the peptide's full array of potential TCR contact residues. Whereas the HLA B*4402-cross-reactive receptor binds amino acids toward the COOH terminus of the peptide, others preferentially favor an NH2-terminal determinant, presumably evading an area that mimics a structure presented on HLA B*4402. Thus, tolerance to a background major histocompatibility antigen can effectively diversify the TCR repertoire for a foreign epitope by deflecting the response away from an immunodominant combination of TCR-binding residues. PMID- 7500018 TI - A rare cryptic translation product is presented by Kb major histocompatibility complex class I molecule to alloreactive T cells. AB - The identity of allogeneic peptide/major histocompatibility complex (MHC) complexes that elicit vigorous T cell responses has remained an interesting problem for both practical and theoretical reasons. Although a few abundant MHC class I-bound peptides have been purified and sequenced, identifying the unique T cell-stimulating peptides from among the thousands of existing peptides is still a very difficult undertaking. In this report, we identified the antigenic peptide that is recognized by an alloreactive bm1 anti-B6 T cell clone using a novel genetic strategy that is based upon measurement of T cell receptor occupancy in single T cells. Using lacZ-inducible T cells as a probe, we screened a splenic cDNA library in transiently transfected antigen-presenting cells (APCs) and isolated a cDNA clone that allowed expression of the appropriate peptide/Kb MHC complex in APC. The antigenic octapeptide (SVVEFSSL) exactly matched the consensus Kb MHC motif, but was surprisingly encoded by a non-ATG defined translation reading frame. Furthermore, the abundance of the naturally processed analog in untransfected cells was estimated to be <10 copies per cell. These results illustrate a novel strategy for identifying T cell-stimulating antigens in general and directly show that alloreactive T cells can respond to rather rare peptide/MHC complexes. These results also suggest that the total pool of processed peptides expressed on the APC surface may include those generated by cryptic translation of normally expressed transcripts. PMID- 7500019 TI - Identification of a CD4+ T cell-stimulating antigen of pathogenic bacteria by expression cloning. AB - Identifying the immunogenic proteins that elicit pathogen-specific T cell responses is key to rational vaccine design. While several approaches have succeeded in identifying major histocompatibility complex (MHC) class I bound peptides that stimulate CD8+ T cells, these approaches have been difficult to extend to peptides presented by MHC class II molecules that stimulate CD4+ T cells. We describe here a novel strategy for identifying CD4+ T cell-stimulating antigen genes. Using Listeria monocytogenes-specific, lacZ-inducible T cells as single-cell probes, we screened a Listeria monocytogenes genomic library as recombinant Escherichia coli that were fed to macrophages. The antigen gene was isolated from the E. coli clone that, when ingested by the macrophages, allowed generation of the appropriate peptide/MHC class II complex and T cell activation. We show that the antigenic peptide is derived from a previously unknown listeria gene product with characteristics of a membrane-bound protein. PMID- 7500017 TI - Activation of virus replication after vaccination of HIV-1-infected individuals. AB - Little is known about the factors that govern the level of HIV-1 replication in infected individuals. Recent studies (using potent antiviral drugs) of the kinetics of HIV-1 replication in vivo have demonstrated that steady-state levels of viremia are sustained by continuous rounds of de novo infection and the associated rapid turnover of CD4+ T lymphocytes. However, no information is available concerning the biologic variables that determine the size of the pool of T cells that are susceptible to virus infection or the amount of virus produced from infected cells. Furthermore, it is not known whether all CD4+ T lymphocytes are equally susceptible to HIV-1 infection at a given time or whether the infection is focused on cells of a particular state of activation or antigenic specificity. Although HIV-1 replication in culture is known to be greatly facilitated by T cell activation, the ability of specific antigenic stimulation to augment HIV-1 replication in vivo has not been studied. We sought to determine whether vaccination of HIV-1-infected adults leads to activation of virus replication and the targeting of vaccine antigen-responsive T cells for virus infection and destruction. Should T cell activation resulting from exposure to environmental antigens prove to be an important determinant of the steady state levels of HIV-1 replication in vivo and lead to the preferential loss of specific populations of CD4+ T lymphocytes, it would have significant implications for our understanding of and therapeutic strategies for HIV-1 disease. To begin to address these issues, HIV-1-infected individuals and uninfected controls were studied by measurement of immune responses to influenza antigens and quantitation of virion-associated plasma HIV-1 RNA levels at baseline and at intervals after immunization with the trivalent influenza vaccine. Influenza vaccination resulted in readily demonstrable but transient increases in plasma HIV-1 RNA levels, indicative of activation of viral replication, in HIV-1-infected individuals with preserved ability to immunologically respond to vaccine antigens. Activation of HIV-1 replication by vaccination was more often seen and of greater magnitude in individuals who displayed a T cell proliferative response to vaccine antigens at baseline and in those who mounted a significant serologic response after vaccination. The fold increase in viremia, as well as the rates of increase of HIV-1 in plasma after vaccination and rates of viral decline after peak viremia, were higher in individuals with higher CD4+ T cell counts.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 7500021 TI - Immunopathology of interleukin (IL) 2-deficient mice: thymus dependence and suppression by thymus-dependent cells with an intact IL-2 gene. AB - Interleukin (IL) 2-deficient mice develop a fatal immunopathology characterized by lymphoadenopathy, splenomegaly, T cell infiltration of the bone marrow, loss of B cells, anemia, and inflammation of the gut. The thymus dependence of these disease symptoms was tested by introducing the IL-2 mutation into athymic mice. With the exception of an increase in CD8+ intrahepatic alpha/beta T cells, IL-2 deficiency had no detectable effect on leukocyte composition or health of athymic mice, indicating a key role for thymus-derived T cells in the initiation of disease and demonstrating that B cell development and survival are independent of IL-2. In adoptive transfer studies, lymph node and spleen cells from euthymic IL 2-deficient mice induced disease in athymic mice with an intact IL-2 gene, suggesting that thymus-independent IL-2-expressing cells are unable to control the development of immune pathology. Both IL-2+ and IL-2-/- bone marrow cells repopulated the thymus and the peripheral T cell compartment of the recombination activator gene 2-deficient recipients, and chimeras that had received IL-2 deficient bone marrow developed immune pathology. Disease development was, however, fully or at least partially prevented when 30% of the bone marrow inoculum was derived from mice able to express IL-2. These results demonstrate that the IL-2 deficiency syndrome depends on the intrathymic differentiation of T cells carrying the IL-2 mutation, and that the abnormal activation of IL-2 deficient lymphocytes can be controlled by thymus-derived but not thymus independent lymphocytes. PMID- 7500020 TI - T helper type 1/T helper type 2 cytokines and T cell death: preventive effect of interleukin 12 on activation-induced and CD95 (FAS/APO-1)-mediated apoptosis of CD4+ T cells from human immunodeficiency virus-infected persons. AB - Human immunodeficiency virus (HIV) infection leads to a progressive loss of CD4+ T helper (Th) type 1 cell-mediated immunity that is associated with defective in vitro CD4+ T cell proliferation and abnormal T cell death by apoptosis in response to T cell receptor (TCR) stimulation. Quantification of interleukin (IL) 2, interferon gamma, IL-4, IL-5, and IL-10 secretion by immunoassays, and of interferon gamma, IL-4 and IL-10 messenger RNA expression by competitive reverse transcriptase polymerase chain reaction after in vitro stimulation of the TCR revealed a similar Th1 cytokine profile in T cells from HIV-infected persons and from controls. These data indicated that the loss of CD4+ Th1 cell function in HIV-infected persons is not related to a Th1 to Th2 cytokine switch as previously proposed, but to a process of activation-induced death of CD4+ Th1 cells. Despite the absence of elevated levels of Th2 cytokines, apoptosis of CD4+ T cells, but not of CD8+ T cells, was prevented in vitro by antibodies to IL-10 or IL-4, two Th2 cytokines that downregulate Th1 cell responses, or by the addition of recombinant IL-12, a cytokine that upregulates Th1 functions. TCR-induced apoptosis of T cell hybridomas and preactivated T cells has been shown to involve the CD95 (Fas/Apo-1) molecule. CD4+ and CD8+ T cells from HIV-infected persons expressed high levels of the CD95 molecule, and, in contrast to T cells from controls, were highly sensitive to antibody-mediated CD95 ligation, which induced apoptosis in a percentage of T cells similar to that induced by TCR stimulation. As TCR-induced apoptosis, CD95-mediated apoptosis of CD4+ T cells, but not of CD8+ T cells, was prevented by the addition of recombinant IL-12. Together, these findings suggest that apoptosis of CD4+ T cells from HIV-infected persons involves an abnormal sensitivity to CD95 ligation, and to TCR stimulation in the presence of normal levels of Th2 cytokines. The preventive effect of IL-12 on both mechanisms has potential implications for the design of immunotherapy strategies aimed at the upregulation of CD4+ Th1 cell functions in AIDS. PMID- 7500022 TI - Metalloproteinase-mediated release of human Fas ligand. AB - Fas ligand (FasL) is a type II integral membrane protein homologous with tumor necrosis factor (TNF). Recent studies indicate that TNF is processed to yield the soluble cytokine by metalloproteinases at the cell surface of activated macrophages and T cells. In the present study, we investigated whether FasL is also released by metalloproteinases. Treatment with hydroxamic acid inhibitors of matrix metalloproteinases specifically led to accumulation of membrane-type FasL (p40) on the surface of human FasL cDNA transfectants and activated human T cells, as estimated by surface immunofluorescence and immunoprecipitation with newly established anti-human FasL monoclonal antibodies. This surface accumulation of mFasL was associated with the decrease of soluble FasL (p27) in the supernatant as estimated by quantitative ELISA and immunoprecipitation with anti-human FasL monoclonal antibodies. These results indicate that human FasL is efficiently released from the cell surface by metalloproteinases like TNF. PMID- 7500023 TI - Thiols decrease human interleukin (IL) 4 production and IL-4-induced immunoglobulin synthesis. AB - N-Acetyl-L-cysteine (NAC) is an antioxidant precursor of intracellular glutathione (GSH), usually given in human as a mucolytic agent. In vitro, NAC and GSH have been shown to act on T cells by increasing interleukin (IL) 2 production, synthesis and turnover of IL-2 receptors, proliferation, cytotoxic properties, and resistance to apoptosis. We report here that NAC and GSH decrease in a dose-dependent manner human IL-4 production by stimulated peripheral blood T cells and by T helper (Th) 0- and Th2-like T cell clones. This effect was associated with a decrease in IL-4 messenger RNA transcription. In contrast, NAC and GSH had no effect on interferon gamma and increased IL-2 production and T cell proliferation. A functional consequence was the capacity of NAC and GSH to selectively decrease in a dose-dependent manner IL-4-induced immunoglobulin (Ig) E and IgG4 production by human peripheral blood mononuclear cells. Interestingly, NAC and GSH also acted directly on purified tonsillar B cells by decreasing the mature epsilon messenger RNA, hence decreasing IgE production. In contrast, IgA and IgM production were not affected. At the same time, B cell proliferation was increased in a dose-dependent manner. Not all antioxidants tested but only SH bearing molecules mimicked these properties. Finally, when given orally to mice, NAC decreased both IgE and IgG1 antibody responses to ovalbumin. These results demonstrate that NAC, GSH, and other thiols may control the production of both the Th2-derived cytokine IL-4 and IL-4-induced Ig in vitro and in vivo. PMID- 7500024 TI - A novel antigen-processing-defective phenotype in major histocompatibility complex class II-positive CIITA transfectants is corrected by interferon-gamma. AB - Presentation of exogenous protein antigens to T lymphocytes is based on the intersection of two complex pathways: (a) synthesis, assembly, and transport of major histocompatibility complex (MHC) class II-invariant chain complexes from the endoplasmic reticulum to a specialized endosomal compartment, and (b) endocytosis, denaturation, and proteolysis of antigens followed by loading of antigenic peptides onto newly synthesized MHC class II molecules. It is believed that expression of MHC class II heterodimers, invariant chain and human leukocyte antigen-DM is both necessary and sufficient to reconstitute a functional MHC class II loading compartment in antigen-presenting cells. Expression of each of these essential molecules is under the control of the MHC class II transactivator CIITA. Unexpectedly, however, whereas interferon gamma stimulation does confer effective antigen-processing function to nonprofessional antigen presenting cells, such as melanoma cells, expression of the CIITA transactivator alone is not sufficient. Activation of antigen-specific T cells thus requires additional CIITA-independent factor(s), and such factor(s) can be induced by interferon gamma. PMID- 7500025 TI - Oncostatin M induces association of Grb2 with Janus kinase JAK2 in multiple myeloma cells. AB - Oncostatin M (OSM) is a 28-kD glycoprotein recently identified as a growth factor for human multiple myeloma cells. It belongs to a family of distantly related cytokines that includes interleukin 6, ciliary neurotrophic factor, leukemia inhibitory factor, and interleukin 11. These cytokines initiate signaling by inducing either homodimerization of gp130 or heterodimerization of gp130 with leukemia-inhibitory factor receptor beta components. Such dimerization in turn activates receptor-associated tyrosine kinases. In the present study using U266B1 human multiple myeloma cells, we show that OSM induces tyrosine phosphorylation and activation of JAK2, but not JAK1 or Tyk2, kinases. The results also demonstrate that OSM induces direct interaction of JAK2 kinase with Grb2, an SH2/SH3 domain containing adaptor protein. The SH2 domain of Grb2 is directly associated with tyrosine-phosphorylated JAK2. Furthermore, the presence of Sos in the JAK2-Grb2 complex suggests a role for Ras in OSM-transduced signaling. PMID- 7500026 TI - Complement-mediated regulation of tissue factor activity in endothelium. AB - Inflammation and immunity may be associated with endothelial cell (EC) injury and thrombus formation. We explored the mechanisms through which a humoral immune response directed against the endothelium might promote coagulation. Using the interaction of anti-EC antibodies and complement (C) with cultured EC as a model, we studied the expression and function of tissue factor, a cofactor for factor VIIa-mediated conversion of factor X to Xa. Exposure of EC to anti-EC antibodies and C in sublytic amounts stimulated the synthesis of tissue factor over a period of 16-42 h. Cell surface expression of tissue factor activity required activation of C and assembly of the membrane attack complex, because expression was inhibited by soluble CR1 and was not detected in the absence of C8. Elaboration of tissue factor messenger RNA was observed over a period of 8-30 h and required protein synthesis. Expression of tissue factor was not a direct consequence of the action of C on the EC but was a secondary response that required as an intermediate step the release of interleukin 1 alpha, an early product of the EC response to C activation. These findings suggest that, after the assembly of membrane attack complex on EC, the production of tissue factor and initiation of coagulation in a blood vessel depend on the production of interleukin 1 alpha and on its availability to stimulate affected EC. PMID- 7500027 TI - Role of the Syk autophosphorylation site and SH2 domains in B cell antigen receptor signaling. AB - To explore the mechanism(s) by which the Syk protein tyrosine kinase participates in B cell antigen receptor (BCR) signaling, we have studied the function of various Syk mutants in B cells made Syk deficient by homologous recombination knockout. Both Syk SH2 domains were required for BCR-mediated Syk and phospholipase C (PLC)-gamma 2 phosphorylation, inositol 1,4,5-triphosphate release, and Ca2+ mobilization. A possible explanation for this requirement was provided by findings that recruitment of Syk to tyrosine-phosphorylated immunoglobulin (Ig) alpha and Ig beta requires both Syk SH2 domains. A Syk mutant in which the putative autophosphorylation site (Y518/Y519) of Syk was changed to phenylalanine was also defective in signal transduction; however, this mutation did not affect recruitment to the phosphorylated immunoreceptor family tyrosine based activation motifs (ITAMs). These findings not only confirm that both SH2 domains are necessary for Syk binding to tyrosine-phosphorylated Ig alpha and Ig beta but indicate that this binding is necessary for Syk (Y518/519) phosphorylation after BCR ligation. This sequence of events is apparently required for coupling the BCR to most cellular protein tyrosine phosphorylation, to the phosphorylation and activation of PLC-gamma 2, and to Ca2+ mobilization. PMID- 7500028 TI - Regulation of the balance of cytokine production and the signal transducer and activator of transcription (STAT) transcription factor activity by cytokines and inflammatory synovial fluids. AB - The balance between type 1 and 2 T helper cell cytokine production plays an important role in several animal models of autoimmunity, and skewed patterns of cytokine expression have been described in human inflammatory diseases. Many cytokines activate signal transducer and activation of transcription (STAT) transcription factors, which, in turn, activate transcription of inflammatory effector genes. We used mononuclear cell priming cultures and inflammatory synovial fluids (SFs) derived from arthritis patients to examine the regulation of cytokine production and STAT activity by an inflammatory synovial microenvironment. Exposure to SFs during priming resulted in an 81% inhibition of interferon (IFN)-gamma, but not interleukin (IL) 4, production by effector cells generated in priming cultures. SF suppression was mediated by IL-4 and IL-10 and inhibition of IL-12 expression, and it was reversed in a dominant fashion by exogenous IL-12. SFs blocked the sustained activity of transcription factor Stat1, but not Stat3, during the priming period, and Stat1 activity was differentially regulated by cytokines in parallel with their positive or negative regulation of IFN-gamma production. Active Stat3, but not Stat1, was detected in cells from inflamed joints. These results suggest a role for altered balance of Stat1 and Stat3 transcriptional activity in the regulation of T cell differentiation and in the pathogenesis of inflammatory synovitis. PMID- 7500029 TI - Superantigen binding to a T cell receptor beta chain of known three-dimensional structure. AB - The three-dimensional structure of an unglycosylated T cell antigen receptor (TCR) beta chain has recently been determined to 1.7 A resolution. To investigate whether this soluble beta chain (murine V beta 8.2J beta 2.1C beta 1) retains superantigen (SAG)-binding activity, we measured its affinity for various bacterial SAGs in the absence of MHC class II molecules. Dissociation constants (KDs) were determined using two independent techniques: surface plasmon resonance detection and sedimentation equilibrium. Specific binding was demonstrated to staphylococcal enterotoxins (SEs) B, C1, C2, and C3 and to streptococcal pyrogenic exotoxin A (SPEA), consistent with the known proliferative effects of these SAGs on T cells expressing V beta 8.2. In contrast, SEA, which does not stimulate V beta 8.2-bearing cells, does not bind the recombinant beta chain. Binding of the beta chain to SAGs was characterized by extremely fast dissociation rates (> 0.1 s-1), similar to those reported for certain leukocyte adhesion molecules. Whereas the beta chain bound SEC1, 2, and 3 with KDs of 0.9 2.5 microM, the corresponding value for SEB was approximately 140 microM. The much weaker binding to SEB than to SEC1, 2, or 3 was surprising, especially since SEB was found to actually be 3- to 10-fold more effective, on a molar basis, than the other toxins in stimulating the parental T cell hybridoma. We interpret these results in terms of the ability of SEC to activate T cells independently of MHC, in contrast to SEB. We have also measured SE binding to the glycosylated form of the beta chain and found that carbohydrate apparently does not contribute to recognition, even though the N-linked glycosylation sites at V beta 8.2 residues Asn24 and Asn74 are at or near the putative SAG-binding site. This result, along with the structural basis for the V beta specificity of SEs, are discussed in relation to the crystal structure of the unglycosylated beta chain. PMID- 7500030 TI - HLA-A2 subtypes are functionally distinct in peptide binding and presentation. AB - Nearly half of HLA-A2-positive individuals in African populations have a subtype of HLA-A2 other than the A*0201 allele. We have isolated the common African HLA A2 subtype genes from Epstein-Barr virus-transformed B cell lines and have established stable class I reduced transfectants expressing these alleles. We have studied the peptide binding and presentation properties of A*0201, A*0202, A*0205, A*0214, and A*6901 by a combination of approaches: assaying direct binding of labeled synthetic peptides, studying the ability of antigen-specific cytotoxic T lymphocytes to recognize peptide-pulsed cells, and sequencing peptide pools and individual ligands eluted from cells. We find that A*0201-restricted peptides can also bind to A*0202 but do not bind strongly to the other alleles in this study. We show that some cytotoxic T lymphocytes can recognize all subtypes capable of binding an antigenic peptide, whereas others are subtype specific. Sequencing of eluted peptides reveals that A*0202 has a similar peptide motif to A*0201, but that A*0205, A*0214, and A*6901 have different motifs. These data strongly support a model in which residue 9 (Phe or Tyr) of the A2/A68/A69 molecules is a critical factor in determining the specificity of the B pocket of the major histocompatibility complex and the position 2 anchor residue of associated peptides. We conclude that a single-amino acid difference in the major histocompatibility complex can be sufficient to cause a dramatic change in the nature of bound peptides, implying that individuals with closely related HLA subtypes may present very different repertoires of antigenic peptides to T cells in an immune response. It is likely to be a general phenomenon that very similar class I subtypes will behave as functionally distinct HLA allotypes. PMID- 7500031 TI - Functional interactions of T cells with endothelial cells: the role of CD40L-CD40 mediated signals. AB - CD40 is expressed on a variety of cells, including B cells, monocytes, dendritic cells, and fibroblasts. CD40 interacts with CD40L, a 30-33-kD activation-induced CD4+ T cell surface molecule. CD40L-CD40 interactions are known to play key roles in B cell activation and differentiation in vitro and in vivo. We now report that normal human endothelial cells also express CD40 in situ, and CD40L-CD40 interactions induce endothelial cell activation in vitro. Frozen sections from normal spleen, thyroid, skin, muscle, kidney, lung, or umbilical cord were studied for CD40 expression by immunohistochemistry. Endothelial cells from all tissues studied express CD40 in situ. Moreover, human umbilical vein endothelial cells (HUVEC) express CD40 in vitro, and recombinant interferon gamma induces HUVEC CD40 upregulation. CD40 expression on HUVEC is functionally significant because CD40L+ Jurkat T cells or CD40L+ 293 kidney cell transfectants, but not control cells, upregulate HUVEC CD54 (intercellular adhesion molecule-1), CD62E (E-selectin), and CD106 (vascular cell adhesion molecule-1) expression in vitro. Moreover, the kinetics of CD40L-, interleukin 1-, or tumor necrosis factor alpha induced CD54, CD62E, and CD106 upregulation on HUVEC are similar. Finally, CD40L CD40 interactions do not induce CD80, CD86, or major histocompatibility complex class II expression on HUVEC in vitro. These results demonstrate that CD40L-CD40 interactions induce endothelial cell activation in vitro. Moreover, they suggest a mechanism by which activated CD4+ T cells may augment inflammatory responses in vivo by upregulating the expression of endothelial cell surface adhesion molecules. PMID- 7500032 TI - The cleavage preference of the proteasome governs the yield of antigenic peptides. AB - Proteasomes degrade endogenous proteins in the cytosol. The potential contribution of the proteasome to the effect of flanking sequences on the presentation of an antigenic epitope presented by the major histocompatibility complex class I allele Ld was studied. Peptides generated in cells from minigenes coding for peptides of 17- and 19-amino acid length were compared with the in vitro 20S proteasome degradation products of the respective synthetic peptides. The quality of generated peptides was independent of ubiquitination. In vivo and in vitro processing products were indistinguishable with respect to peptide size and abundance. Altering the neighboring sequence substantially improved the yield of the final antigenic nonapeptide by 20S proteasome cleavage. These results suggest that, in addition to the presence of major histocompatibility complex class I allelic motifs, the cleavage preference of the proteasome can define the antigenic potential of a protein. PMID- 7500033 TI - Release of the mucosal mast cell granule chymase, rat mast cell protease-II, during anaphylaxis is associated with the rapid development of paracellular permeability to macromolecules in rat jejunum. AB - The soluble granule chymase, rat mast cell protease-II (RMCP-II), is abundantly expressed in intestinal mucosal mast cells (MMC) but its function is not known. One hypothesis is that RMCP-II degrades the epithelial basement membrane and promotes the loss of enterocytes typically associated with type I hypersensitivity reactions in the rat. To test this hypothesis more directly, ex vivo perfusion of the cranial mesenteric artery and jejunal lumen was used to monitor the anaphylactic release of RMCP-II and its effects on mucosal permeability and epithelial integrity. Within 2 min of intravascular challenge with soluble adult Nippostrongylus brasiliensis worm antigen there was a 1,000 fold (P < 0.02) increase in the concentration of RMCP-II in the vascular perfusate from the jejunum of Nippostrongylus-sensitized rats but not the controls. Similarly, translocation of RMCP-II into the gut lumen increased 10 fold (P < 0.02) after 2 min only in worm antigen-challenged immune rats. Using an identical protocol, but incorporating Evans blue-labeled human serum albumin (EB HSA) in the vascular perfusate, the timing of the release of RMCP-II into the two compartments was very similar to the first experiment and furthermore the translocation of EB-HSA increased 18-fold (P < 0.05) after 4 min in sensitized rats challenged with worm antigen. To examine the effects of RMCP-II more directly 1 mg of the highly purified chymase was introduced into the cranial mesenteric artery in ex vivo perfused normal rats. A significant (P < 0.05) 70 fold increase in concentration of RMCP-II in jejunal perfusate occurred after 6 min. In a repeat dose-response experiment, infusion of 0.375, 0.75, or 1.5 mg of RMCP-II, together with EB-HSA, established that the cumulative amounts of RMCP-II and EB-HSA translocated from the vasculature to the gut lumen in each perfusion (during the 10-min period of RMCP-II infusion) were significantly correlated. Analysis of intestinal perfusates by SDS-PAGE and by Western blotting using monoclonal anti-RMCP-II antibody confirmed that there was a concomitant translocation of both the protease and EB-HSA into the gut lumen. Histological evaluation of the mucosa failed to reveal any significant morphological change in any of the experiments. The rapid development of macromolecular leak, its association with the translocation of RMCP-II, and the absence of gross epithelial lesions, suggest for the first time that a mast cell granule chymase increases epithelial permeability via a paracellular route and implies that the substrate may be a protein, or proteins, in the epithelial junctional complex. PMID- 7500034 TI - The peptide-binding motif for the human transporter associated with antigen processing. AB - Presentation of antigenic peptides by human leukocyte antigen class I molecules is dependent on peptide transport into the endoplasmic reticulum by the transporters associated with antigen processing (TAP) (Germain, R. N. 1994. Cell. 76:287-299). This translocation step is currently regarded as permissive for all peptides with COOH-terminal residues capable of binding to HLA class I molecules (Momburg, F., J. Roelse, J. C. Howard, G. W. Butcher, G.J. Hammerling, and J.J. Neefjes. 1994. Nature (Lond.). 367:648-651). In this report, we show that the human transporter selects peptides according to a binding motif based on the strong effects on peptide affinity of the three NH2-terminal positions and the COOH-terminal residues. TAP favors strongly hydrophobic residues in position 3 (P3) and hydrophobic or charged residues in P2, whereas aromatic or acidic residues in P1, as well as Pro in P1 and P2, have strong deleterious effects. Selection of naturally presented peptides by the transporter is suggested by their higher average affinity for TAP, as compared to nonselected peptides. The TAP preferences in the three NH2-terminal positions correspond to those of the vast majority of human leukocyte antigen class I alleles, but they represent an obstacle for peptide supply to some alleles, e.g., the B7-like group. We propose that peptides binding to these alleles, and in general, peptides with TAP affinities below a certain threshold, may be transported as extended precursors. PMID- 7500035 TI - Immunoreactivity for interleukin 3 and 5 and granulocyte/macrophage colony stimulating factor of intestinal mucosa in bronchial asthma. AB - T lymphocytes and eosinophils are important components of the inflammatory cell infiltrate in bronchial mucosa in asthma. Because activated lymphocytes migrate through the thoracic duct and the general circulation to remote glandular and mucosal sites, we initiated this study to evaluate pathological abnormalities and immunoreactivity for interleukin (IL) 3, IL-5, and granulocyte/macrophage colony stimulating factor (GM-CSF) of intestinal mucosa in bronchial asthma. 15 asthmatic patients, 8 nonasthmatic patients with chronic obstructive pulmonary disease, 6 atopic nonasthmatic healthy controls, and 6 nonatopic healthy controls were studied. Duodenal biopsies were performed by endoscopy. A significantly increased number of intraepithelial lymphocytes and eosinophils and a significant accumulation of mononuclear cells (lymphocytes and mast cells) and eosinophils in the lamina propria were detected in asthmatics and atopic controls. Immunostaining with antibodies directed against IL-3, IL-5, and GM-CSF was positive in asthmatics and atopic controls, whereas no staining was observed in nonatopic controls and chronic obstructive pulmonary disease. Combined ultrastructural study and immunogold labeling demonstrated that IL-3, IL-5, and GM-CSF were localized in eosinophils and mast cells. Although devoid of gastrointestinal symptoms, asthmatics and asymptomatic atopics had duodenal pathological abnormalities mimicking those observed in the bronchial mucosa in asthma, suggesting that the whole mucosal immune system is involved in bronchial asthma. PMID- 7500036 TI - Altered hepatic transport of immunoglobulin A in mice lacking the J chain. AB - We have created J chain knockout mice to define the physiologic role of the J chain in immunoglobulin synthesis and transport. The J chain is covalently associated with pentameric immunoglobulin (Ig) M and dimeric IgA and is also expressed in most IgG-secreting cells. J chain-deficient mice have normal serum IgM and IgG levels but markedly elevated serum IgA. Although polymeric IgA was present in the mutant mice, a larger proportion of their serum IgA was monomeric than was found in wild-type mouse serum. Bile and fecal IgA levels were decreased in J chain-deficient mice compared with wild-type mice, suggesting inefficient transport of J chain-deficient IgA by hepatic polymeric immunoglobulin receptors (pIgR). The pIgR-mediated transport of serum-derived IgA from wild-type and mutant mice was assessed in Madin-Darby canine kidney (MDCK) cells transfected with the pIgR. These studies revealed selective transport by pIgR-expressing MDCK cells of wild-type IgA but not J chain-deficient IgA. We conclude that although the J chain is not required for IgA dimerization, it does affect the efficiency of polymerization or have a role in maintaining IgA dimer stability. Furthermore, the J chain is essential for efficient hepatic pIgR transport of IgA. PMID- 7500037 TI - TAP-independent, beta 2-microglobulin-dependent surface expression of functional mouse CD1.1. AB - CD1 molecules consist of beta 2-microglobulin (beta 2m) noncovalently complexed to a non-major histocompatibility complex (MHC)-encoded monomorphic integral membrane protein homologous to MHC class I alpha chains. Little is known about the requirements for cell surface expression and T cell recognition of CD1. We inserted the mouse CD1.1 gene into vaccinia virus to create a recombinant virus expressing CD1.1 under the control of a viral promoter. Using this recombinant virus to infect normal or mutant cell lines, we found that the expression of molecules reactive with the CD1.1-specific monoclonal antibody 3C11 requires the expression of beta 2m but was not affected by the absence of the MHC-encoded peptide transporter (TAP). Consistent with these results, IL-2 production by the mCD1.1-specific T cell hybridoma DN32.D3 was induced by thymocytes from normal mice or mice with a homozygous deletion of the TAP1 gene, but not by thymocytes from mice with a homozygous deletion of the beta 2m gene. These results indicate that expression of functional mCD1.1 occurs in a beta 2m-dependent, TAP independent manner. PMID- 7500038 TI - Ontogenic development and tissue distribution of V gamma 1-expressing gamma/delta T lymphocytes in normal mice. AB - A hamster monoclonal antibody (mAb) recognizing an epitope in the V gamma 1-J gamma 4-C gamma 4 chain of the gamma/delta T cell receptor has been generated. Using this mAb, we have quantitated the occurrence of V gamma 1-bearing gamma/delta T cells in the developing thymus and in the lymphoid organs and several epithelia of adult mice. The V gamma 1-expressing cells constitute a minor gamma/delta T cell subpopulation during fetal and early postnatal life, but they constitute a major population of gamma/delta T cells in the thymus and in the peripheral lymphoid organs in adult mice. In addition, we found that V gamma 1-bearing cells comprise a large proportion (15-60%) of the gamma/delta T cells present in the intestinal epithelium (i-IEL) in all strains of mice tested. V gamma 1+ i-IEL are present in athymic (nude) mice and in antigen-free mice, demonstrating that they can develop extrathymically and that their presence in the intestinal epithelium is independent of the antigenic load of the gut. Our results show that V gamma 1-bearing lymphocytes account for the largest population of gamma/delta T cells in the mouse. This population includes a thymus dependent component that homes to the secondary lymphoid organs and a thymus independent component that constitutes a major fraction of the gamma/delta i IELs. PMID- 7500039 TI - A role for stem cell factor and c-kit in the murine intestinal tract secretory response to cholera toxin. AB - The role of stem cell factor (SCF) and its receptor (c-kit) in the intestinal secretory response to cholera toxin (CT) was investigated using a ligated intestinal loop model in mice having mutations in the dominant white spotting (W) locus and the steel (Sl) locus. W/Wv mice, which express an aberrant form of the c-kit protein, failed to give an intestinal secretory response after luminal CT challenge. In contrast, W/Wv mice and their control littermates had equivalent intestinal secretory responses to Escherichia coli heat-stable enterotoxin (STa). Sl/Sld mice, which express only a soluble truncated form of SCF, also gave a significantly reduced intestinal secretory response to CT when compared to the secretory response of their littermate controls. The unresponsiveness of W/Wv mice to CT was restricted to the intestinal tract since these mice had foot pad swelling responses to CT challenge that were equivalent to their littermate controls. Restoration of mast cells in W/Wv mice by bone marrow transplantation of control littermate bone marrow did not reverse the CT-unresponsiveness of the intestinal tract. Histological evaluation of the gastrointestinal tract from W/Wv mice showed a normal distribution of enterochromaffin cells (ECC). CT challenge of either ligated intestinal loops from C57B1/6 mice or a mouse intestinal epithelial cell line (MODE-K) resulted in elevated levels of mRNA for SCF. MODE-K cells exposed to CT also had enhanced expression of c-kit. Finally, fluid obtained from CT-challenged ligated intestinal loops from C57B1/6 mice contained significant levels of SCF. Taken together, the above results suggest that CT induced intestinal secretory responses are dependent upon SCF-c-kit interactions. These interactions appear to be induced as a consequence of CT stimulation of the intestinal tract and may also play a role in the development or functionality of the enteric nervous system. PMID- 7500040 TI - Anti-immunoglobulin M activates nuclear calcium/calmodulin-dependent protein kinase II in human B lymphocytes. AB - We and others have previously shown that the nuclear protein, Ets-1, is phosphorylated in a calcium-dependent manner after ligation of immunoglobulin (Ig) M on B lymphocytes. As this phosphorylation was independent of protein kinase C activity, we tested whether a calcium/calmodulin-dependent protein kinase (CaM kinase) might phosphorylate the Ets-1 protein after elevation of intracellular free calcium concentrations. The dephosphorylated form of Ets-1 has been shown to bind to chromatin, suggesting that the operative kinase should be detectable in the nucleus. We prepared nuclear extracts from two human B cell lines in which increased intracellular free calcium levels correlated with increased phosphorylation of the Ets-1 protein. Activity of the CaM kinases was determined using a synthetic peptide substrate both in the absence and presence of an inhibitor specific for the CaM kinase family, KN-62. Stimulation of cells with anti-IgM led to increased activity of a nuclear kinase that could phosphorylate the peptide, and this activity was reduced by 10 microM KN-62. Kinase activity was reduced in lysates preadsorbed using an antibody specific for CaM kinase II. Two-dimensional phosphopeptide maps of the Ets-1 protein from cells incubated with ionomycin or anti-IgM contained two unique phosphopeptides that were absent in untreated cells. Incubation of isolated Ets-1 protein with purified CaM kinase II produced phosphorylation of peptides that migrated identically to those found in cells incubated with either anti-IgM or ionomycin. These data suggest a model of signal transduction by the antigen receptor on B lymphocytes in which increased intracellular free calcium can rapidly activate nuclear CaM kinase II, potentially resulting in phosphorylation and regulation of DNA-binding proteins. PMID- 7500041 TI - Abnormal glucocorticoid receptor-activator protein 1 interaction in steroid resistant asthma. AB - Glucocorticosteroids are a very effective treatment for asthma and other chronic inflammatory diseases. However, a small proportion of patients is resistant to the therapeutic effects of glucocorticoids. Pharmacokinetic and ligand binding studies suggest that the molecular abnormality in steroid resistance lies distal to nuclear translocation. We have previously reported that there is a decreased ability of glucocorticoid receptors (GR) to bind to the DNA-binding site in peripheral blood mononuclear cells (PBMC) after dexamethasone treatment. This reduced DNA binding was due to a decrease in the number of receptors available rather than an alteration in affinity for DNA. To study this reduced DNA binding, we examined the ability of the nuclear translocated transcription factors activator protein 1 (AP-1), nuclear factor kappa B (NF-kappa B) and cyclic AMP response element-binding protein (CREB) to bind to their DNA-binding sites and to interact with GR in PBMC from patients with steroid-sensitive and steroid resistant asthma. There was a significant reduction in the interaction between GR and AP-1 in these steroid-resistant patients, although interaction with other transcription factors activated in inflammation (NF-kappa B and CREB) was unaffected. An increase in the basal levels of AP-1 DNA binding was also detected in the nuclei from steroid-resistant asthmatic patients. There were no differences in the amount of messenger RNA detected for the components of AP-1, c Fos and c-Jun, nor in the sequences of these messenger RNAs. These results suggest either that the ability of the GR to bind to glucocorticoid response elements and AP-1 is altered in steroid-resistant patients or that increased levels of AP-1 prevent GR DNA binding, and that this may be the molecular basis of resistance to the antiinflammatory effect of steroids in these cells. PMID- 7500043 TI - Bcl-XL displays restricted distribution during T cell development and inhibits multiple forms of apoptosis but not clonal deletion in transgenic mice. AB - The survival of T lymphocytes is tightly controlled during development. Here, we show that Bcl-xL, a protein homologue of Bcl-2, is highly regulated in the thymus in a pattern different than that of Bcl-2. The maximum expression was in CD4+CD8+ thymocytes, a developmental stage where Bcl-2 is downregulated. To assess the role of Bcl-xL in thymocyte apoptosis, we generated mice overexpressing an E mu bcl-x transgene within the T cell compartment. Constitutive expression of Bcl-xL resulted in accumulation of thymocytes and mature T cells in lymphoid organs. Thymocytes overexpressing Bcl-xL exhibited increased viability in vitro and were resistant to apoptosis induced by different signals, including glucocorticoid, gamma irradiation, calcium ionophore, and CD3 cross-linking. However, Bcl-xL was unable to block clonal deletion of thymocytes reactive with self-superantigens or H-Y antigen. These studies demonstrate that Bcl-2 and Bcl-xL, two functionally related proteins, are regulated independently during T cell development. In contrast to Bcl-2, which has been implicated in the maintenance of mature T cells, Bcl-xL appears to provide a survival signal for the maintenance of more immature CD4+CD8+ thymocytes before positive selection. PMID- 7500044 TI - Expression of costimulatory molecules B7-1 (CD80), B7-2 (CD86), and interleukin 12 cytokine in multiple sclerosis lesions. AB - Resting autoreactive T cells are present in the circulation of normal individuals without pathologic consequences. In autoimmune animal models, stimulation of these self-reactive T cells in the presence of costimulatory molecules B7-1 results in T cell-mediated autoimmune disease, whereas B7-2 stimulation generates regulatory autoreactive T cells that abrogate disease severity. Thus, reactivation in the brain of myelin-autoreactive T cells by antigen with costimulatory molecules may be a critical event in the pathophysiology of multiple sclerosis (MS), a putative autoimmune disease of central nervous system (CNS) myelin. We investigated the expression of cytokines and costimulatory molecules in a panel of 41 histologically characterized CNS specimens from 15 MS and 10 control cases using semiquantitative reverse transcriptase-polymerase chain reaction and immunocytochemistry. In four cases, vascular CNS infarcts with inflammation were compared with MS plaques from the same brain. We observed increased expression of B7-1 and interleukin (IL) 12p40 in acute MS plaques, particularly from early disease cases but not in inflammatory infarcts. B7-1 staining was localized predominantly to the lymphocytes in perivenular inflammatory cuffs but not the parenchyma. In contrast, B7-2 was expressed predominantly on macrophages both in MS lesions of varied time duration and in inflammatory infarcts. These findings indicate that an early event in the initiation of MS involves upregulation of B7-1 and IL-12, resulting in conditions that maximally stimulate T cell activation and induction of T helper 1-type immune responses. PMID- 7500042 TI - On the pathogenesis of atherosclerosis: enzymatic transformation of human low density lipoprotein to an atherogenic moiety. AB - Combined treatment with trypsin, cholesterol esterase, and neuraminidase transforms LDL, but not HDL or VLDL, to particles with properties akin to those of lipid extracted from atherosclerotic lesions. Single or double enzyme modifications, or treatment with phospholipase C, or simple vortexing are ineffective. Triple enzyme treatment disrupts the ordered and uniform structure of LDL particles, and gives rise to the formation of inhomogeneous lipid droplets 10-200 nm in diameter with a pronounced net negative charge, but lacking significant amounts of oxidized lipid. Enzymatically modified LDL (E-LDL), but not oxidatively modified LDL (ox-LDL), is endowed with potent complement activating capacity. As previously found for lipid isolated from atherosclerotic lesions, complement activation occurs to completion via the alternative pathway and is independent of antibody. E-LDL is rapidly taken up by human macrophages to an extent exceeding the uptake of acetylated LDL (ac-LDL) or oxidatively modified LDL. After 16 h, cholesteryl oleate ester formation induced by E-LDL (50 micrograms/ml cholesterol) was in the range of 6-10 nmol/mg protein compared with 3-6 nmol/mg induced by an equivalent amount of acetylated LDL. At this concentration, E-LDL was essentially devoid of direct cytotoxic effects. Competition experiments indicated that uptake of E-LDL was mediated in part by ox LDL receptor(s). Thus, approximately 90% of 125I-ox-LDL degradation was inhibited by a 2-fold excess of unlabeled E-LDL. Uptake of 125I-LDL was not inhibited by E LDL. We hypothesize that extracellular enzymatic modification may represent an important step linking subendothelial deposition of LDL to the initiation of atherosclerosis. PMID- 7500045 TI - Physical dissociation of the TCR-CD3 complex accompanies receptor ligation. AB - Recent studies indicate that there may be functional uncoupling of the TCR-CD3 complex and suggest that the TCR-CD3 complex is composed of two parallel signal transducing units, one made of gamma delta epsilon chains and the other of zeta chains. To elucidate the molecular mechanisms that may explain the functional uncoupling of TCR and CD3, we have analyzed their expression by using flow cytometry as well as immunochemical means both before and after stimulation with anti-TCR-beta, anti-CD3 epsilon, anti-CD2, staphylococcal enterotoxin B, and ionomycin. We present evidence that TCR physically dissociates from CD3 after stimulation of the TCR-CD3 complex. Stimulation with anti-CD3 resulted in down modulation of TCR within 45 min whereas CD3 epsilon was still expressed on the cell surface as detected by flow cytometry. However, the cell surface expression of TCR and CD3 was not affected when cells were stimulated with anti-TCR-beta under the same conditions. In the case of anti-CD3 treatment of T cells, the TCR down-modulation appeared to be due to the internalization of TCR, as determined by immunoelectron microscopy. Immunochemical analysis of cells after stimulation with either anti-TCR or anti-CD3 mAbs revealed that the overall protein levels of TCR and CD3 were similar. More interestingly, the dissociation of the TCR-CD3 complex was observed with both treatments and occurred in a manner that the TCR and the associated TCR-zeta chain dissociated as a unit from CD3. These results provide the first report of physical dissociation of TCR and CD3 after stimulation through the TCR-CD3 complex. The results also suggest that the signal transduction pathway triggered by TCR may differ from that induced by CD3. PMID- 7500046 TI - A pathway of costimulation that prevents anergy in CD28- T cells: B7-independent costimulation of CD1-restricted T cells. AB - A class of molecules that is expressed on antigen presenting cells, exemplified by CD80 (B7), has been found to provide a necessary costimulatory signal for T cell activation and proliferation. CD28 and CTLA4 are the B7 counterreceptors and are expressed on the majority of human CD4+ T cells and many CD8+ T cells. The signal these molecules mediate is distinguished from other costimulatory signals by the finding that T cell recognition of antigen results in a prolonged state of T cell unresponsiveness or anergy, unless these costimulatory molecules are engaged. However, nearly half of the CD8+ and CD4-CD8- T cells lack CD28, and the costimulatory signals required for the activation of such cells are unknown. To understand the pathways of activation used by CD28- T cells, we have examined the costimulatory requirements of antigen-specific CD4-CD8- TCR(+)-alpha/beta circulating T cells that lack the expression of CD28. We have characterized two T cell lines, DN1 and DN6, that recognize a mycobacterial antigen, and are restricted not by major histocompatibility complex class I or II, but by CD1b or CD1c, two members of a family of major histocompatibility complex-related molecules that have been recently implicated in a distinct pathway for antigen presentation. Comparison of antigen-specific cytolytic responses of the DN1 and DN6 T cell lines against antigen-pulsed CD1+ monocytes or CD1+ B lymphoblastoid cell lines (B-LCL) demonstrated that these T cells recognized antigen presented by both types of cells. However, T cell proliferation occurred only when antigen was presented by CD1+ monocytes, indicating that the CD1+ monocytes expressed a costimulatory molecule that the B-LCL transfectants lacked. This hypothesis was confirmed by demonstrating that the T cells became anergic when incubated with the CD1(+)-transfected B-LCL in the presence of antigen, but not in the absence of antigen. The required costimulatory signal occurred by a CD28-independent mechanism since both the CD1+ monocytes and CD1+ B-LCL transfectants expressed B7 1 and B7-2, and DN1 and DN6 lacked surface expression of CD28. We propose that these data define a previously unrecognized pathway of costimulation for T cells distinct from that involving CD28 and its counterreceptors. We suggest that this B7-independent pathway plays a crucial role in the activation and maintenance of tolerance of at least a subset of CD28- T cells. PMID- 7500047 TI - Inflammatory microcrystals differentially regulate the secretion of macrophage inflammatory protein 1 and interleukin 8 by human neutrophils: a possible mechanism of neutrophil recruitment to sites of inflammation in synovitis. AB - Human neutrophils at inflammatory sites may be an important source of the chemotactic cytokines macrophage inflammatory protein 1 alpha (M1P-1 alpha; a C-C chemokine) and interleukin 8 (IL-8; a C-X-C chemokine). In this study, we show that the inflammatory microcrystals monosodium urate monohydrate (MSU) and calcium pyrophosphate dihydrate (CPPD), the major mediators of gout and pseudogout, differentially regulate the production of these two chemokines by human neutrophils. Both MSU and CPPD increased the secretion of IL-8 by neutrophils in a dose- and time-dependent manner, but had no effect on that of MIP-1 alpha. Since inflammatory cytokines are likely to be present in the synovium during crystal-induced inflammation, we examined the interaction between TNF-alpha and GM-CSF and the crystals. Both TNF-alpha and GM-CSF stimulated IL-8 production; however, only TNF-alpha exerted a significant effect on MIP-1 alpha secretion in neutrophils. IL-8 production induced by TNF-alpha and GM-CSF was synergistically enhanced in the presence of MSU or CPPD, whereas MIP-1 alpha secretion induced by TNF was completely inhibited in the presence of either MSU or CPPD. Interestingly, no interaction between the crystals and the inflammatory cytokines was observed with respect to synthesis of the C-X-C chemokine MGSA in neutrophils. These results suggest that the combination of TNF-alpha and GM-CSF with MSU or CPPD will lead to the production of IL-8 by neutrophils and abolish the release of MIP-1 alpha, an event that will theoretically lead to recruitment of neutrophils but not mononuclear cells. These results are in accordance with the pathological state of gout and pseudogout, where the predominant inflammatory cell is the neutrophil. PMID- 7500049 TI - Improved engraftment of human hematopoietic cells in severe combined immunodeficient (SCID) mice carrying human cytokine transgenes. AB - We have generated immunodeficient scid-/scid- (SCID)-transgenic mice expressing the genes for human interleukin 3, granulocyte/macrophage-colony stimulating factor, and stem cell factor. We have compared engraftment and differentiation of human hematopoietic cells in transgenic SCID mice with two strains of nontransgenic SCID mice. Human bone marrow cells carrying the CD34 antigen or human umbilical cord blood were injected into sublethally irradiated recipients. Human DNA was detected by polymerase chain reaction in peripheral blood and bone marrow of 14 of 28 transgenic SCID mice after transplantation, but in only 2 of 15 nontransgenic SCID littermates at a 10-fold lower level. Bone marrow cultures 8 wk after transplantation of cord blood gave rise to human burst-forming unit erythroid, colony-forming unit granulocyte/macrophage, or granulocyte/erythroid/macrophage/megakaryocyte colonies. Engraftment was observed for up to 6 mo in transgenic SCID mice, twice as long as nontransgenic littermates or previous studies in which transplanted SCID mice were given daily injections of growth factors. We conclude that the level and duration of engraftment of human cells in SCID mice can be improved by expression of human cytokine transgenes and that transgenic SCID mice are an efficient model system for the study of human hematopoiesis. PMID- 7500048 TI - Inhibition of invariant chain (Ii)-calnexin interaction results in enhanced degradation of Ii but does not prevent the assembly of alpha beta Ii complexes. AB - Calnexin is a resident protein of the endoplasmic reticulum (ER) that associates with nascent protein chains. Among the newly synthesized integral membrane proteins known to bind to calnexin is invariant chain (Ii), and Ii release from calnexin coincides with proper assembly with major histocompatibility complex (MHC) class II heterodimers. Although calnexin association with several membrane glycoproteins depends on interactions involving N-linked glycans, we previously reported that a truncation mutant of mouse Ii (mIi1-107) lacking both N glycosylation sites was highly effective in associating with MHC class II heterodimers and escorting these dimers through the secretory pathway. This could indicate that calnexin, despite binding to both Ii and class II, is not necessary for the proper interaction of these proteins, or that in contrast to most membrane glycoproteins, the N-linked glycans of Ii are not critical to its interaction with this chaperone. To examine this issue, we have directly explored the binding of calnexin to both Ii truncation mutants lacking the typical sites of N-glycosylation or Ii produced in cells treated with tunicamycin to prevent glycan addition. These experiments revealed that either method of eliminating N linked carbohydrates on Ii also inhibited association with calnexin. A lumenally truncated form of Ii (mIi1-131) that still has N-linked carbohydrates showed a decreased affinity for calnexin compared with intact Ii, however, indicating that calnexin-Ii binding is not determined solely by the sugar moieties. All forms of Ii lacking N-linked sugars and showing defective association with calnexin also had enhanced rates of preendosomal degradation. Despite this effect on degradation rate, tunicamycin treatment did not inhibit the association of class II with glycan-free Ii. These data support the view that calnexin is not an absolute requirement for the proper assembly of class II-Ii nonamers, but rather acts primarily to retain Ii in the ER and to inhibit its degradation. These two properties of calnexin-Ii interaction may help ensure that sufficient intact Ii is available for efficient inactivation of the binding sites of newly synthesized class II molecules, while limiting the ability of excess free Ii to alter the transport properties of the early endocytic pathway. PMID- 7500050 TI - Low levels of HIV-1 infection in cutaneous dendritic cells promote extensive viral replication upon binding to memory CD4+ T cells. AB - Earlier work has identified a cell population that replicates HIV-1 in the absence of standard T cell stimuli. The system consists of dendritic cells and memory T lymphocytes that emigrate from organ cultures of human skin and together support a productive infection with HIV-1. These emigrants resemble cells that can be found in mucous membranes and that normally traffic in afferent lymph. Here, we report that a low level of infection in the dendritic cell can initiate extensive HIV-1 replication in cocultures with T cells. First we extended our earlier work to larger skin specimens from cadavers. As long as the organ cultures were set up within 36 h of death, the emigrant leukocytes were comparable to cells from fresh surgical specimens in number, phenotype, and function. These mixtures of dendritic cells and T cells provided the milieu for a productive infection with several virus isolates. When purified dendritic cells were separately pulsed with virus and then mixed with T cells that had not been pulsed with HIV-1, active infection ensued. The infectivity of HIV-pulsed dendritic cells persisted for at least 1.5 d in culture, but was blocked if AZT was added during that time to block reverse transcription in the dendritic cells. The number of copies of proviral DNA in the dendritic cells corresponded to < 100 copies per 5 X 10(4) cells, but upon mixing with T cells, > 10(4) copies were found 5-7 d later. By contacting syngeneic T cells, extralymphoid depots of dendritic cells--even with a low viral burden as has been reported in vivo--may contribute to chronic HIV-1 replication in infected individuals. PMID- 7500051 TI - Successful ultraviolet B treatment of psoriasis is accompanied by a reversal of keratinocyte pathology and by selective depletion of intraepidermal T cells. AB - Skin irradiation with ultraviolet B (UVB) is a common and often durable treatment for psoriasis and other inflammatory skin disorders. We studied the effects of UVB on keratinocytes and leukocytes in psoriatic tissue and in culture. In nine patients treated repetitively, most of the cellular and molecular changes that typify the psoriatic epidermis reverted to normal. Keratinocyte hyperplasia, assessed by expression of the Ki-67 cell cycle antigen, decreased by 70%, and residual cell proliferation was appropriately confined to the basal layer. Epidermal thickening was reduced by 60%, and a granular layer reformed. Expression of keratin 16, as well as suprabasal integrin alpha 3 and insulin-like growth factor-1 receptors, was eliminated, whereas filagrin increased markedly. UVB also depleted > 90% of the CD3+, CD8+, and CD25+ T cells from the psoriatic epidermis, whereas dermal T cells were only minimally depressed. The latter finding parallels the known inability of these doses of UVB to penetrate the dermis. In tissue culture, UVB was antiproliferative and cytotoxic toward T cells and keratinocytes, but the T cells were 10-fold more sensitive. Furthermore, low doses of UVB induced apoptosis in lymphocytes but not keratinocytes, as detected by the TUNEL (TdT-mediated dUTP-biotin nick end labeling) technique. The selective effects of UVB on intraepidermal T cells in situ and in culture support the hypothesis that epidermal alterations in psoriasis can be normalized by a depletion of activated intraepidermal T cells. PMID- 7500052 TI - gro-beta, a -C-X-C- chemokine, is an angiogenesis inhibitor that suppresses the growth of Lewis lung carcinoma in mice. AB - We have found that two chemokines, recombinant gro-alpha and gro-beta, specifically inhibit growth factor-stimulated proliferation of capillary endothelial cells in a dose-dependent manner, whereas gro-gamma has no inhibitory effect. In vivo, gro-beta inhibits blood vessel formation in the chicken chorioallantoic membrane assay. It is sufficiently potent to effectively suppress basic fibroblast growth factor-induced corneal neovascularization after systemic administration in mice. Further, gro-beta significantly inhibits the growth of murine Lewis lung carcinoma in syngeneic C57B16/J and immunodeficient nude mice without toxicity. In vitro, Lewis lung carcinoma cells are completely insensitive to recombinant gro-beta at high concentrations that significantly inhibit endothelial cell proliferation. This finding supports the conclusion that gro beta inhibits Lewis lung tumor growth by suppression of tumor-induced neovascularization. PMID- 7500054 TI - Positive selection of mouse NK1+ T cells by CD1-expressing cortical thymocytes. AB - Mouse NK1+ T cells constitute a subset of alpha/beta TCR+ T cells that specialize in the rapid production of cytokines, in particular IL-4, and may promote the differentiation of Th2-type CD4 T cells. Their TCRs, like those of a homologous subset of human T cells, use an invariant TCR alpha chain and were recently shown to be specific for the beta 2-microglobulin-associated, MHC class I-like CD1 molecules, which are encoded outside the MHC. In contrast to mainstream thymocytes, which recognize their positively selecting MHC ligand on thymic epithelial cells, positive selection of NK1+ T cells requires their CD1 ligand to be expressed on bone marrow-derived cells. To investigate the nature of the bone marrow-derived cell involved, chimeric mice were constructed with tissues from normal, SCID, and MHC-deficient mice, so that CD1 could be selectively expressed by different subsets of bone marrow-derived cells in the thymus. CD1 expression was also directly assessed using an anti-CD1 mAb, and a CD1-specific T cell hybridoma. The results suggest that immature (CD4+8+ double-positive) cortical thymocytes are the source of CD1 presentation for positive selection of NK1+ T cells. PMID- 7500053 TI - T cell receptor- and beta 1 integrin-mediated signals synergize to induce tyrosine phosphorylation of focal adhesion kinase (pp125FAK) in human T cells. AB - The beta 1 subfamily of integrins is thought to play an important role in both the adhesion/migration and proliferation/differentiation of T cells. beta 1 integrins can provide T cell costimulation through interaction of very late antigen (VLA) 4 (VLA-4) (alpha 4 beta 1) and VLA-5 (alpha 5 beta 1) with the extracellular matrix protein fibronectin (FN), or by VLA-4 binding to its cell surface ligand, vascular cell adhesion molecule (VCAM) 1. The mechanism by which beta 1 integrin members transduce T cell-costimulatory signals is poorly understood. Studies in non-T cells have demonstrated regulation of the tyrosine focal adhesion kinase pp125FAK by beta 1 integrin engagement and, most recently, indicate a role for pp125FAK in linking integrin-mediated signal transduction to the Ras pathway (Schaller, M. D., and J. T. Parsons, 1994, Curr. Opin. Cell. Biol. 6: 705-710; Schlaepfer, D. D., S. K. Hanks, T. Hunter, and P. Van der Geer. 1994. Nature (Lond.), 372:786-790). Although pp125FAK kinase occurs in T cells, there are no reports on its regulation in this cell type. The studies described in this article characterize novel regulation of pp125FAK by the T cell receptor (TCR)-CD3 antigen complex and beta 1 integrins, and provide the first account, in any cell type, of integrin alpha 4 beta 1-mediated pp125FAK tyrosine phosphorylation. We demonstrate a rapid and sustained synergistic increase in tyrosine phosphorylation of human pp125FAK in Jurkat T cells after simultaneous (a) triggering of the TCR-CD3 complex, and (b) alpha 4 beta 1 and alpha 5 beta 1 integrin-mediated binding of these cells to immobilized FN or alpha 4 beta 1 integrin-mediated binding to immobilized VCAM-1. Studies with normal peripheral blood-derived CD4+ human T blasts confirm the synergistic action of a TCR-CD3 complex-mediated costimulus with a FN- or VCAM-1-dependent signal in the induction of T cell pp125FAK tyrosine phosphorylation. In vitro kinase assays performed on pp125FAK immunoprecipitates isolated from Jurkat cells and normal CD4+ T cells identified a coprecipitating 57-kD tyrosine-phosphorylated protein (pp57), distinct from pp59fyn or pp56lck. These results indicate, for the first time, the involvement of a specific kinase, pp125FAK, in alpha 4 beta 1- and alpha 5 beta 1-mediated T cell-costimulatory signaling pathways. In addition, the data demonstrate novel regulation of pp125FAK tyrosine phosphorylation by the TCR CD3 complex. PMID- 7500056 TI - Fluconazole therapy for candiduria. PMID- 7500055 TI - The expression and regulation of nitric oxide synthase in human osteoarthritis affected chondrocytes: evidence for up-regulated neuronal nitric oxide synthase. AB - Classically, osteoarthritis (OA) has been considered a noninflammatory disease. However, the detection of selected inflammatory mediators in osteoarthritic fluid, in the absence of significant inflammatory cell infiltrate, is increasingly appreciated. We sought to identify the inflammatory component in human OA-affected cartilage that may be involved in cartilage damage/destruction. Using Western blot analysis and an antibody to the conserved region of nitric oxide synthase (NOS), we have observed up-regulation of NOS, one of the "key players" of inflammation, in chondrocytes of OA-affected patients. Remarkably, none of the cartilage samples examined from normal joints demonstrated detectable amounts of this NOS. Western blot analysis using the same alpha-NOS antibody indicated that this NOS from OA-affected cartilage (OA-NOS) was larger in size than (and distinct from) transfected human hepatocyte or murine inducible NOS (iNOS) (150 versus 133 kD) and similar in size to neuronal constitutive NOS (ncNOS). Antibodies specific for iNOS showed binding to murine and human iNOS but not to OA-NOS, endothelial constitutive NOS, or ncNOS. Antibodies specific for ncNOS bound to ncNOS and also to OA-NOS, but not to murine or human iNOS or endothelial constitutive NOS. Incubation of OA cartilage in serum-free medium resulted in spontaneous release, for up to 72 h, of substantial amounts of nitrite (up to approximately 80 microM/100 mg wet tissue), which could be inhibited by at least 80% with various inhibitors of iNOS, including inhibitors of protein synthesis and transcription factor NF-kappa B, but which (unlike murine macrophage iNOS) was not sensitive to hydrocortisone or TGF-beta. Exposure of OA-affected cartilage to interleukin 1 beta, tumor necrosis factor-alpha, and lipopolysaccharide resulted in approximately 20-50% augmentation of nitrite accumulation, which was also sensitive to cycloheximide and pyrrolidine dithiocarbamate. Hence, our data indicate that OA-NOS (based on immunoreactivity and molecular weight) is similar to ncNOS and that it releases nitric oxide, which may contribute to the inflammation and pathogenesis of cartilage destruction in OA. PMID- 7500057 TI - Prostate cancer screening. PMID- 7500058 TI - FPs and emergency medicine. PMID- 7500061 TI - From the land of milk and honey. PMID- 7500060 TI - Advanced obstetrical training for FPs. PMID- 7500059 TI - Minocycline vs tetracycline. PMID- 7500062 TI - Mental health problems within primary care: shooting first and then asking questions? PMID- 7500063 TI - Psychological management by family physicians. AB - BACKGROUND: It is frequently assumed that primary care physicians seldom provide psychological interventions to their patients with mental health problems. This study examines self-reports of psychological interventions by family physicians. METHODS: Primary care patients (N = 937) completed a mental health screening form immediately prior to their medical visit. Results were withheld from their seven respective physicians. Following the visit, the physicians were asked to classify the range of psychological interventions they used to manage their patients' emotional problems during the visit. A structured psychiatric diagnostic interview was subsequently administered to a subgroup of the patients (n = 388). RESULTS: At least one psychological intervention was provided to nearly one fourth (24.1%) of the patients. The interventions included listening to the patient's emotional problems (22.4%), providing advice (19.0%), discussing the patient's mental disorder diagnosis (11.4%), and providing individual counseling (8.4%) or family counseling (0.6%). Two thirds (66.7%) of the patients who reported that their emotional health was poor received at least one of these psychological interventions. In a multivariate model, the likelihood of receiving a psychological intervention was higher for patients who were separated or divorced; those between 45 and 59 years of age; those with less than a college education; those who received disability payments; those who reported poor emotional health; and those who had a positive screening result for panic disorder, major depressive disorder, or obsessive-compulsive disorder. CONCLUSIONS: Primary care physicians may be far more extensively involved in providing psychological interventions than is commonly assumed. PMID- 7500064 TI - Correlates of depression in primary care. AB - BACKGROUND: Depression is a significant health problem in the United States. This study examined the relationship between depression and substance use, substance problems, conduct disorders, and sociodemographic factors in primary care settings. METHODS: A survey of 1898 patients in 88 primary care offices was conducted using a self-administered health-habits questionnaire. Depression was assessed for both lifetime and for the past 30 days using the Diagnostic and Statistical Manual of Mental Disorders, Third Edition Revised (DSM-III-R) criteria. RESULTS: A total of 21.7% of women and 12.7% of men met DSM-III-R criteria for depression in the 30 days prior to completing the survey. Lifetime rates of depression were 36.1% for women and 23.3% for men. Young women who smoke, drink, or use marijuana, and both men and women with antisocial personality disorder and a family history of mental health problems are particularly at high risk for depression. CONCLUSIONS: One in 5 women and one in 10 men who see their primary care physicians have recently been depressed. PMID- 7500065 TI - A strategy to arrest and reverse coronary artery disease: a 5-year longitudinal study of a single physician's practice. AB - BACKGROUND: Animal experiments and epidemiological studies have suggested that coronary disease could be prevented, arrested, or even reversed by maintaining total serum cholesterol levels below 150 mg/dL (3.88 mmol/L). In 1985, we began to study how effective one physician could be in helping patients achieve this cholesterol level and what the associated effect of achieving and maintaining this cholesterol level has on coronary disease. METHODS: The study included 22 patients with angiographically documented, severe coronary artery disease that was not immediately life threatening. These patients took cholesterol-lowering drugs and followed a diet that derived no more than 10% of its calories from fat. Disease progression was measured by coronary angiography and quantified with the percent diameter stenosis and minimal lumen diameter methods. Serum cholesterol was measured biweekly for 5 years and monthly thereafter. RESULTS: Of the 22 participants, 5 dropped out within 2 years, and 17 maintained the diet, 11 of whom completed a mean of 5.5 years of follow-up. All 11 of these participants reduced their cholesterol level from a mean baseline of 246 mg/dL (6.36 mmol/L) to below 150 mg/dL (3.88 mmol/L). Lesion analysis by percent stenosis showed that of 25 lesions, 11 regressed and 14 remained stable. Mean arterial stenosis decreased from 53.4% to 46.2% (estimated decrease = 7%; 95% confidence interval [CI], 3.3 to 10.7, P < .05). Analysis by minimal lumen diameter of 25 lesions found that 6 regressed, 14 remained stable, and 5 progressed. Mean lumen diameter increased from 1.3 mm to 1.4 mm (estimated increase = 0.08 mm; 95% CI, -0.06 to 0.22, P = NS). Disease was clinically arrested in all 11 participants, and none had new infarctions. Among the 11 remaining patients after 10 years, six continued the diet and had no further coronary events, whereas the five dropouts who resumed their prestudy diet reported 10 coronary events. CONCLUSIONS: A physician can influence patients in the decision to adopt a very low-fat diet that, combined with lipid-lowering drugs, can reduce cholesterol levels to below 150 mg/dL and uniformly result in the arrest or reversal of coronary artery disease. PMID- 7500066 TI - Subgaleal hematoma: the need for increased awareness of risk. AB - Subgaleal hematoma, also known as subaponeurotic hemorrhage, is a serious complication of birth that is associated with vacuum-assisted delivery. Despite a high rate of mortality associated with subgaleal hematoma, it has received relatively little attention in the medical literature. Lack of awareness may lead to delayed diagnosis and serious consequences for infants. This paper is a report of six cases and a literature review. Prevention and early recognition and treatment of the condition can occur only with increased practitioner awareness of this entity. PMID- 7500068 TI - A family practice article filing system. AB - An organized article filing system can be a timesaver for the busy clinician. A comprehensive, specialty-based filing system for the family physician is presented. The full system uses about 700 three-tab manila folders. Each major medical specialty heading is placed on an empty center-tab folder. Subfolders within each major heading are all right- or left-tab folders, alternating for each successive major heading. Topics that overlap specialty are cross referenced. Additional major headings are included for presenting symptoms, procedures, practice management topics, society membership, licensure, etc. The resulting filing system is intuitive to use since articles are filed by the specialty to which the the topic is most closely related. Journal articles and associated patient education materials may be filed in the same folder for quick retrieval. PMID- 7500067 TI - Office laboratory diagnosis of vaginitis. Clinician-performed tests compared with a rapid nucleic acid hybridization test. AB - BACKGROUND: The traditional diagnosis of vaginitis incorporates patient symptoms, clinical findings observed during vaginal examination, and laboratory analysis of vaginal fluid. The purpose of this study was to evaluate routine clinician performed office laboratory diagnostic techniques for women with abnormal vaginal symptoms, and to compare these results with those obtained by a DNA hybridization test for Trichomonas vaginalis, Gardnerella vaginalis, and Candida species. METHODS: The study included 501 symptomatic women who were between the ages of 14 and 67 years. Three vaginal specimens were obtained for saline wet mount, potassium hydroxide (KOH) prep, amine "sniff", pH, and nucleic acid hybridization (T vaginalis, G vaginalis, and Candida sp) tests. Clinicians and medical technicians independently evaluated the wet mount, KOH prep, amine, and pH tests. A medical technician processed the DNA tests according to manufacturer's protocol. RESULTS: Of 499 subjects for whom complete data were available, vulvovaginal candidiasis was diagnosed in 20.0%, vaginal trichomoniasis in 7.4%, and bacterial vaginosis in 52.1%. Fourteen percent of subjects had multiple vaginal infections. The sensitivity and specificity of clinician microscopically diagnosed vulvovaginal candidiasis, vaginal trichomoniasis, and bacterial vaginosis were 39.6% and 90.4%, 75.0% and 96.6%, and 76.5% and 70.8%, respectively. The sensitivity and specificity of the DNA probe diagnosis of the same types of vaginitis were 75.0% and 95.7%, 86.5% and 98.5%, and 95.4% and 60.7%, respectively. When only women with multiple vaginal infections were considered, the percentages of correct clinician diagnoses for vulvovaginal candidiasis, vaginal trichomoniasis, and bacterial vaginosis were 49.3%, 83.6%, and 59.7%, respectively. For the DNA probe test, the percentages of correct diagnoses were 72.9%, 92.9%, and 90.0%, respectively. CONCLUSIONS: Primary care clinicians demonstrated a high specificity but low sensitivity when identifying vaginal trichomoniasis and vulvovaginal candidiasis by microscopic techniques. Correct microscopic diagnosis of bacterial vaginosis was even more difficult for clinicians, as was the diagnosis of multiple vaginal infections. Clinicians were not as accurate as the DNA probe test in diagnosing vaginal infections. Clinicians need more education in the laboratory diagnosis of vaginitis. Clinicians should carefully scrutinize each microscopic slide, systematically examine the slide for each type of vaginitis, and consider specimen pH and the presence of leukocytes, Lactobacillus organisms, or amine odor as additional clues to infection. PMID- 7500069 TI - Nefazodone and venlafaxine: two new agents for the treatment of depression. AB - Nefazodone and venlafaxine are two new antidepressants with pharmacologic actions and side-effect profiles that differ from those of the selective serotonin reuptake inhibitors (SSRIs). In addition to inhibition of serotonin reuptake, nefazodone exhibits 5-HT2 antagonism. Venlafaxine inhibits the reuptake of both norepinephrine and serotonin. Both appear to be effective and well tolerated in the treatment of major depressive disorder. Nefazodone differs from the SSRIs in that it does not cause sexual dysfunction or sleep impairment. Venlafaxine differs from the SSRIs because of its effects augmenting norepinephrine, as well as its demonstrated efficacy in the treatment of severely depressed, melancholic patients. Both drugs appear to hold significant promise as effective and well tolerated medications for the treatment of major depression in the primary care setting. PMID- 7500070 TI - An update of vitamin B12 metabolism and deficiency states. AB - Vitamin B12 deficiency may be underestimated in the general population. High-risk groups for the deficiency syndrome include the elderly, patients taking ulcer medications over long periods, patients with acquired immunodeficiency syndrome, vegetarians, patients who have undergone stomach resection or small bowel resection, or both, and patients with dementia. The vitamin B12 deficiency syndrome is characterized by five stages, the fifth of which results in irreversible neuropsychiatric manifestations. Although the deficiency is easily treated, diagnosis is somewhat complicated by the shortcomings of the various tests. Current state-of-the-art testing uses serum cobalamin levels as a screening test and serum or urine homocysteine and methylmalonic acid determinations as confirmatory tests. Vitamin B12 deficiency is treatable with monthly injections, large doses of daily oral supplement tablets, or an intranasal gel, which is far better absorbed than comparable oral supplements. PMID- 7500071 TI - Cement burn. AB - People working with cement should be aware of the potential for cement burn. The alkalinity of cement is due to the exothermic reaction of calcium oxide with water, which forms calcium hydroxide. Prolonged exposure of the skin to wet cement may result in a caustic burn. PMID- 7500072 TI - Treating unstable angina. PMID- 7500073 TI - Venous thromboembolism. PMID- 7500074 TI - Bacterial vaginosis. PMID- 7500075 TI - Who are we? We are against violence. PMID- 7500076 TI - Staying loose. PMID- 7500077 TI - Violence in America. Reaching epidemic proportions. PMID- 7500078 TI - Abuse of the elderly. AB - American society is growing ever more violent. The elderly are not exempted from this epidemic despite legislation in all 50 states to protect vulnerable adults. It is unclear whether the incidence of elder abuse is actually rising; certainly the number of elderly and reports of abuse are rising. Case reports of elder abuse only first appeared in the medical literature some 20 years ago. The crime is especially heinous as the perpetrator is in a position of trust, similar to child abuse. It took society years to recognize the serious problem of child abuse. Elder abuse seems doomed to repeat that ponderous course. Like child abuse, elder mistreatment tends to be episodic and recurrent rather than an isolated event. It tends to escalate and rarely spontaneously resolves unless major environmental changes occur. In fact, elderly abuse is only slightly less common than child abuse. States on average spend about $22 per child for youth protective services, but only $2.90 per elder for their protective services, although some 40% of reported abuse involves elders. PMID- 7500081 TI - Physical abuse of the elderly. AB - Physical abuse of the elderly is a frequent occurrence within the older population. For many reasons, including lack of recognition for the entity in the medical literature, physical abuse of the elderly is difficult to recognize. The injuries of physical abuse may be nondescript, but patterned injuries, injuries in unusual anatomic locations and injuries in various stages of healing should alert the practicing physician to the possibility of physical abuse of the elderly. PMID- 7500079 TI - Child abuse. AB - Child abuse or maltreatment is well documented in the United States. The Department of Health and Human Services and the American Medical Association estimate that at least 1 million incidents of physical, emotional and/or sexual maltreatment occur each year; child abuse is reported to cause 2,500 to 5,000 deaths annually. A 1990 study found that approximately 22.6 out of every 1,000 children in the United States experienced abuse or neglect, yet only half of these incidents were reported to child protective service agencies. Organizations and resources for physicians dealing with child abuse are provided in Table 1. PMID- 7500080 TI - Epidemic of women battering. Recognition and intervention by health care professionals. AB - Domestic violence impacts every member of society and takes an emotional toll on the victim, her family (especially children), friends, and law enforcement and health professionals called on to assist. It affects the economic system in medical expenses, social services to protect the victim and her children, time lost from work by the victim, and expense to jail and prosecute the abuser. PMID- 7500083 TI - Blue collar doctor. PMID- 7500082 TI - NIH research. An investment in prevention. AB - The goal of prevention research is to promote and preserve the health of all Americans, which in turn maintains and improves the health of the nation. The National Institutes of Health supports prevention research and related activities in many innovative and challenging areas. The knowledge gained from this research has resulted in lifestyle changes for millions of Americans who now can look forward to longer, healthier and more productive lives. In addition, prevention research and related activities represent our best opportunity to reduce the ever increasing portion of our resources spent on treating preventable illness. PMID- 7500084 TI - Good science rules in the controversy surrounding breast implants. PMID- 7500086 TI - The role of copper in the pMMO of Methylococcus capsulatus bath: a structural vs. catalytic function. AB - Methanotrophs convert methane to methanol by the methane monooxygenase (MMO). It is well known that two forms of the MMO can be expressed: one form is found in the cytoplasm, or in the soluble fraction (sMMO); the other is associated with the membranes, or particulate fraction (pMMO). The sMMO has been extensively examined, and much is known about its structure and mechanism of dioxygen activation. The pMMO, however, is less well understood: the enzyme has proven difficult to purify as it loses activity once the membranes become solubilized. Furthermore, although copper is known to be important for the stability and activity of the pMMO, its role is still unclear. In a recent study, we reported the use of electron paramagnetic resonance (EPR) spectroscopy to probe the nature of the copper ions in these membranes. Two EPR signals were uncovered for the highly oxidized membranes: one set of signals arises from the type 2 Cu(II) centers, and the other has been assigned to trinuclear copper (Cu(II) clusters (H.-H. Nguyen, et al., J. Biol. Chem., 269, 14995 (1994). Here, we attempt to correlate the EPR spectra of the membrane fraction of Methylococcus capsulatus Bath with the amount of copper present in the membranes and to the activity of the pMMO as measured by the production of propylene oxide from propene by the pMMO. From these studies, we conclude that the primary role of copper is in the active site of the pMMO rather than simply a structural one. PMID- 7500085 TI - Reactive arthritis. PMID- 7500087 TI - Synthesis and structures of Zn(C6H12OS2)2(ClO4)2 and Zn(C3H6NS2)2(C3H4N2)--model compounds for the Zn sites in RNA polymerase. AB - Model compounds for the Zn sites of the beta' and the beta subunits in RNA polymerase [1] were synthesized. Single crystal structures and X-ray absorbtion spectroscopy measurements for these two model complexes are reported. In Zn(C6H12OS2)2(ClO4)2, the Zn is coordinated by four sulfur and two oxygen atoms. The average Zn-S bond length is 2.514 A and the Zn-O bond length is 2.089 A, which are similar to these bond distances reported for the Zn site in the beta' subunit of RNA polymerase. In Zn(C3H6NS2)2(C3H4N2), the Zn atom is coordinated by four sulfur atoms and one nitrogen atom of an imidazole group. The average of the Zn-S bond length is 2.469 A and the Zn-N bond length is 2.009 A, which are also similar to the Zn-S and Zn-N bonds in the beta subunits of RNA polymerase. PMID- 7500088 TI - The recharacterization of a polysaccharide iron complex (Niferex). AB - An oral hematinic marketed as "Niferex," the active component of which is a polysaccharide-iron complex (PIC), has recently been recharacterized. PIC is synthesized by the neutralization of an FeCl3 carbohydrate solution. Original characterization of this complex by Mossbauer spectroscopy and X-ray powder diffraction suggested that the iron-rich core was similar in structure to the mineral ferrihydrite. Higher precision X-ray powder diffraction now indicates that the core has a long-range order more similar to the mineral akaganeite, beta FeOOH, than to ferrihydrite. This structure has been found for other similar ferric iron-carbohydrate polymers, especially those synthesized by the hydrolysis of FeCl3. Also discussed are the variable temperature (24-295 K) Mossbauer spectroscopic data for PIC. The first example of EXAFS data for polysaccharide iron complexes confirms that the iron is in an octahedral environment, coordinated to oxygen, with a short-range order similar to that for ferritin. The second iron shells in the PIC samples are less ordered than the second shell in ferritin. The size of the PIC core was found to be approximately 5 nm by X-ray powder diffraction, and is of the same order of magnitude as the ferritin core. PMID- 7500089 TI - Spectroscopic, magnetic, and electrochemical behavior of the copper(II) complex of carnosine. AB - Different physicochemical studies were undertaken with polycrystalline samples of the complex [Cu2(carnosine)2(H2O)2].2H2O. The infrared spectrum was discussed in comparison with that of free carnosine and on the basis of the known structural data. Magnetic susceptibility measurements were performed between 4.2 and 300 K, showing an effective magnetic moment of 1.79 BM. Both the electronic (reflectance) and ESR spectra were compatible with the existence of a dx2-y2 ground state. The axial reversed ESR spectrum could be explained on the basis of a very weak interdimeric coupling mechanism. The electrochemical behavior, investigated by cyclic voltammetry, shows that the complex possesses a very high redox stability. The possible SOD-like activity was tested using the NBT/superoxide reduction assay. The results show a negligible SOD activity. PMID- 7500090 TI - A Mossbauer and X-ray powder diffraction study of some ferrous hematinics. AB - Iron deficiency anemia is a relatively common illness that can arise from a number of different causes. Three ferrous salts are usually used in its treatment: ferrous fumarate, gluconate, and sulfate. They are administered orally and are relatively well tolerated. These hematinics have been studied by Mossbauer spectroscopy and X-ray powder diffraction, and can easily be distinguished by both techniques. It was found that the two ferrous sulfates studied (Eckerd and SmithKline Beckman Co.) most closely resemble the monohydrate by comparison of the X-ray powder pattern with those of the JCPDS. Both the ferrous fumarate (Femiron) and gluconate (Spring Valley) had approximately 10% ferric iron present. To the authors' knowledge, this is the first reported Mossbauer spectrum for ferrous fumarate. PMID- 7500091 TI - Superoxide dismutase mimetic activity of macrocyclic Cu(II) tetraanhydroaminobenzaldehyde (TAAB) complex. AB - The superoxide dismutase mimetic activity of a copper(II) complex with the tetraazamacrocyclic ligand TAAB has been investigated using the xanthin-xanthine oxidase system for generation of superoxide. The complex exhibits 50% inhibition of the 3-(4-iodophenyl)-2-(4-nitrophenyl)-5-phenyltetrazolium chloride (INT) reduction at the concentration of 0.144 mumol.l-1 (pH 7.8, 25 degrees C). The activity survives in the presence of chelators including EDTA and serum albumin. The enzymatic activity of the complex represents about 3% of that of intact Cu2Zn2SOD. PMID- 7500092 TI - Alzheimer's disease: a treatment in sight? PMID- 7500093 TI - Investigations of disorders of balance. PMID- 7500095 TI - The silent period induced by transcranial magnetic stimulation in muscles supplied by cranial nerves: normal data and changes in patients. AB - The silent period induced by transcranial magnetic stimulation of the sensorimotor cortex (Magstim 200, figure of eight coil, loop diameter 7 cm) in active muscles supplied by cranial nerves (mentalis, sternocleidomastoid, and genioglossus) was studied in 14 control subjects and nine patients with localised lesions of the sensorimotor cortex. In the patients, measurements of the silent period were also made in the first dorsal interosseus and tibialis anterior muscles. In the controls, there was a silent period in contralateral as well as ipsilateral cranial muscle and the duration of the silent period increased with increasing stimulus intensities. The mean duration of the silent period was around 140 ms in contralateral mentalis muscle and around 90 ms in contralateral sternocleidomastoid muscle at 1.2 x threshold stimulation strengths. Whereas the duration of the silent period in ipsilateral mentalis muscle was shorter than on the contralateral side it was similar on both sides in sternocleidomastoid muscle. In patients with focal lesions of the face associated primary motor cortex and corresponding central facial paresis, the silent period in mentalis muscle was shortened whereas it was unchanged or prolonged in limb muscles (first dorsal interosseus, tibialis anterior) with stimulation over the affected hemisphere. By contrast, in a patient with a lesion within the parietal cortex, the silent period in mentalis muscle was prolonged with stimulation of the affected side. PMID- 7500096 TI - Clinical and [18F] dopa PET findings in early Parkinson's disease. AB - Twenty seven patients with recent onset (mean symptom duration 22 (SD 14) months, Hoehn and Yahr score 1.8 (SD 0.7)) Parkinson's disease were studied with [18F]dopa PET. There was a correlation between putamen influx (Ki) and clinical rating, but not symptom duration. In 11 patients with hemi-Parkinson's disease of recent onset there were significant differences between normal (mean 0.0123 (SD 0.0023)), asymptomatic (mean 0.0099 (0.0020)) and symptomatic (mean 0.0070 (00.014)) putamen Kis. This suggests that Parkinson's disease has a widely variable rate of progression, and is most compatible with a short preclinical period. Symptom onset was estimated at a putamen Ki of between 57% and 80% of normal. Most ipsilateral putamen Ki values in early asymmetric Parkinson's disease fell within the normal range. The implication is that either the disease is not established in the ipsilateral putamen or that the technique is insufficiently sensitive to detect it. Discriminant analysis completely separated the normal and Parkinson's disease cohorts, but when a discriminant function from a previous study was used predictively four of the 27 patients with Parkinson's disease were incorrectly classified as normal. PMID- 7500094 TI - New phenotype of the cerebral autosomal dominant arteriopathy mapped to chromosome 19: migraine as the prominent clinical feature. AB - A survey was carried out on a large family presenting the symptoms of familial arteriopathy (CADASIL) recently mapped to chromosome 19. This is characterised clinically by recurrent subcortical infarcts developing into pseudobulbar palsy and subcortical dementia, and radiologically by early MRI abnormalities. To characterise this familial condition, 43 members older than 20 years and spreading over four generations were studied clinically (31 living, 12 deceased), genetically, and radiologically by MRI (n = 31). Twenty out of 43 were found to be clinically symptomatic and of these 13 out of 31 had MRI abnormalities. Genetic studies mapped this condition to the locus of CADASIL (lod score > 3). The natural history suggests a chronological clinicoradiological staging of this phenotype of CADASIL: stage I between 20 and 40 years with frequent migraine-like episodes and well delineated lesions of the white matter; stage II between 40 and 60 years with stroke-like episodes, bipolar or monopolar-like psychotic disorders, coalescent lesions of the white matter, and well delineated lesions of the basal ganglia; and stage III over 60 years with subcortical dementia, pseudobulbar palsy, diffuse leukoencephalopathy, and multiple well delineated lesions of the basal ganglia. This phenotype differs from the other two previously described by high frequency of migraine, frequency of psychotic disorders, and early neurological manifestations. The new acronym "cerebral autosomal dominant arteriopathy with subcortical infarcts, leukoencephalopathy, and migraine" (CADASILM) is proposed to better describe this particular subvariety of CADASIL. PMID- 7500098 TI - Mentha piperita (peppermint). PMID- 7500097 TI - Botulinum toxin F in the treatment of torticollis clinically resistant to botulinum toxin A. AB - Two reports have shown a Japanese preparation of botulinum toxin type F (BTX-F) to be an effective alternative for patients with torticollis who develop clinical resistance to botulinum toxin type A (BTX-A). A group of patients with torticollis, comprising five secondary non-responders and one primary non responder, were treated with a preparation of BTX-F produced in the UK (Speywood Pharmaceuticals). A low dose of BTX-F (220 mouse units (MU) in total) was given into clinically affected neck muscles, followed six weeks later by an injection of a total of 520 MU. Antibodies to BTX-A (mouse protection assay) were present in all secondary non-responders but not in the primary non-responder. No patients developed atrophy after injection of Dysport BTX-A (40 MU) into the left extensor digitorum brevis muscle whereas pronounced atrophy occurred in all patients after injection of 40 MU of BTX-F into the right extensor digitorum brevis muscle. Three patients improved subjectively after treatment with 220 MU BTX-F and five (all secondary non-responders) after the subsequent dose of 520 MU (two considerably), with reduced Tsui scores, but group scores were only significantly changed after the higher dose. The primary non-responder remained unchanged after both doses of BTX-F. One patient reported mild dysphagia with 520 MU BTX-F. Mean duration of improvement with 520 MU BTX-F was five (range 4-6)weeks. Thus BTX-F provides benefit for BTX-A non-responders with few side effects but for a shorter period than BTX-A, possibly due to relative underdosing. As with BTX-A, biological sensitivity to BTX-F does not necessarily predict a clinical response. PMID- 7500101 TI - The Sophy valve and the el-Shafei shunt system for adult hydrocephalus. AB - A selected series of 22 adult patients with hydrocephalus were treated by a shunt system incorporating a variable pressure Sophy valve or by ventriculojugular shunting against the direction of blood flow using the El-Shafei system. One patient had insertion of two Sophy valves and an El-Shafei shunt. Patient selection was reserved to those with hydrocephalus thought to be at high risk when shunted with systems containing a conventional unipressure valve. None of the eight patients who had ventriculojugular shunting by the El-Shafei method demonstrated any notable clinical or radiological improvement subsequent to shunt insertion. Of the 16 Sophy devices inserted only seven produced a satisfactory result. The current evaluation of shunt malfunction could be improved by support for a national register. PMID- 7500100 TI - Embolisation of recently ruptured intracranial aneurysms. AB - Patients with subarachnoid haemorrhage due to the rupture of aneurysms unsuitable for craniotomy and clipping have been treated by coil embolisation within three weeks. Sixty nine of 75 consecutive patients were successfully treated. Procedure related complications occurred in 10 patients, resulting in permanent neurological deficits in three and one death (4.8%). The Glasgow outcome scores at six weeks were 53 grade 1, seven grade 2, four grade 3, and five grade 5. These results are comparable with surgical series despite a high proportion of aneurysms in the posterior cerebral circulation. PMID- 7500102 TI - Cerebral haemodynamics and long-term prognosis after extracranial-intracranial bypass surgery. AB - The effectiveness of extracranial-intracranial arterial (EC-IC) bypass surgery for haemodynamic stroke remains controversial. In this study correlation of presurgical and postsurgical cerebral haemodynamics with long term prognosis was evaluated. Twenty eight patients (25 men, three women: mean age 61.4 (SD 8.2) years) with reduced cerebrovascular reserve due to steno-occlusive cerebral vascular disease formed the study group. Measurement of mean hemispheric cerebral blood flow (mCBF) and the cerebral vasodilatory capacity (% mCVR) with an intravenous acetazolamide injection were performed by a xenon-133 inhalation method and SPECT. Patients were treated with EC-IC bypass surgery and measurements of mCBF and % mCVR were made again about one month after surgery. The patients were then observed for a long period (range 27-115 months). During the observation period, four patients experienced subsequent ischaemic strokes. The presurgical and the postsurgical resting mCBF for the affected hemisphere were significantly reduced in the patients with strokes during follow up compared with the values in patients without strokes during follow up (P < 0.03 and 0.01 respectively). The % mCVR of the affected hemisphere was significantly raised after surgery in all patients except one (P < 0.01). The postsurgical change in resting mCBF was not unidirectional. In conclusion, resting mCBF was unchanged and % mCVR was improved after EC-IC bypass surgery in patients with reduced cerebrovascular reserve. The group of patients with a reduced presurgical and postsurgical resting mCBF continue to be a high risk group for subsequent ischaemic stroke seen after EC-IC bypass surgery. PMID- 7500099 TI - Heterogeneous cerebral glucose metabolism in normal pressure hydrocephalus. AB - The regional cerebral metabolic rate for glucose (rCMRglu) has never been investigated in large consecutive groups of patients with normal pressure hydrocephalus (NPH), a potentially treatable form of dementia with an unpredictable outcome after shunt surgery. Using PET and 18F-2 fluorodeoxyglucose, rCMRglu was studied in 18 patients who fulfilled hydrodynamic criteria for NPH and in whom a biopsy of the frontal cortex was obtained. When compared with an age matched group of 11 healthy subjects, the patients with NPH showed a significant rCMRglu reduction in all cortical and subcortical regions of interest. Individual metabolic patterns, however, disclosed a large topographical heterogeneity. Furthermore, histopathological examination identified Alzheimer's disease or cerebrovascular disease in six cases, and no parenchymal disease or non-specific degenerative processes in the remaining 12. After separating the patients according to the histological diagnosis, the rCMRglu patterns were still heterogeneous, the abnormalities ranging from focal to diffuse in both subgroups. After shunt operation, 11 patients did not improve or worsened clinically. Six patients improved; of those, two had Alzheimer changes and two cerebrovascular changes in their biopsy. The metabolic pattern of these six patients did not differ from the rest of the NPH group. The results indicate that the NPH syndrome may be non-specifically associated with different degenerative disorders. The metabolic heterogeneity, together with the heterogeneous histopathological findings, indicate the necessity of reevaluating the pathogenesis of the NPH syndrome, and may account for the high variability in the success rate of shunt surgery series. PMID- 7500103 TI - Eyelid "apraxia" in patients with motor neuron disease. AB - Three patients with motor neuron disease had eyelid "apraxia" with impaired voluntary but preserved involuntary eyelid movements. Attempts were made to localise the lesions responsible with neuroimaging and neuropathological examination. PMID- 7500104 TI - Headache. PMID- 7500105 TI - Secondary cluster headache responsive to sumatriptan. AB - A patient was seen with secondary cluster headache whose acute pain responded promptly to sumatriptan. The headaches started after injury to the vertebral artery. This finding provides clinical affirmation of the existence of the trigeminal/cervical nuclear overlap that is central to this condition. PMID- 7500106 TI - Prognostic value of the amount of post-traumatic subarachnoid haemorrhage in a six month follow up period. AB - Clinical and radiological patterns from 148 patients with post-traumatic subarachnoid haemorrhage (TSAH) were analysed with specific regard for the amount and distribution of blood in subarachnoid spaces to verify if these variables have any influence on overall outcome. The degree and extent of TSAH were classified according to Fisher's criteria: in 93 patients it was grade 1, in 36 grade 2, in 13 grade 3, and in six grade 4. There was a significant correlation between increasing clinical severity at admission and the amount of subarachnoid bleeding and a direct relation between a favourable outcome and a low Fisher grade. The distribution of subarachnoid blood was not significantly related to clinical condition at admission, whereas the pattern had a significant impact on the outcome. The results of the present study confirm that TSAH is a negative prognostic factor. Whereas the degree of TSAH is mainly related to clinical conditions at admission, the presence of subarachnoid blood clots both in basal cisterns and over the cerebral convexity indicates a poor outcome. PMID- 7500107 TI - Neurofibromatosis type 1 in childhood: correlation of MRI findings with intelligence. AB - In a group of 28 children with neurofibromatosis type 1 aged between 4 and 16 years, neuroradiological findings were correlated with intelligence as measured by the Wechsler scales. The presence or specific location in the brain of T2 weighted prolonged signals on MRI was not associated with cognitive problems. No other physical characteristics associated with neurofibromatosis type 1 were found to correlate significantly with IQ. At the present these T2 weighted hyper intense spots should not be used to predict neurofibromatosis type 1 associated cognitive problems. PMID- 7500108 TI - Swelling at the site of a skull defect during migraine headache. PMID- 7500109 TI - Thalamic stimulation for severe action tremor after lesion of the superior cerebellar peduncle. PMID- 7500111 TI - Unilateral proptosis due to cerebellar stroke. PMID- 7500110 TI - Distal myopathy as the presenting manifestation of sarcoidosis. PMID- 7500112 TI - Normal frontal cortex histology and immunohistochemistry in patients with motor neuron disease. PMID- 7500113 TI - Very severe amnesia with acute onset after isolated hippocampal damage due to systemic lupus erythematosus. PMID- 7500114 TI - Lamotrigine control of idiopathic trigeminal neuralgia. PMID- 7500116 TI - Proceedings of the joint summer meeting of the British Neuropsychiatric Association and the British Association for Psychopharmacology, Cambridge, 15-17 July 1995. PMID- 7500115 TI - Clinical evaluation of vasospasm in subarachnoid haemorrhage by in vivo microdialysis. PMID- 7500119 TI - European Stroke Prevention Study 2: Baseline data. PMID- 7500118 TI - Suspected triphenyltin poisoning. PMID- 7500117 TI - Acute necrotising encephalopathy of childhood presenting with multifocal, symmetric brain lesions occurring outside Japan. PMID- 7500120 TI - The kinocilium of auditory hair cells and evidence for its morphogenetic role during the regeneration of stereocilia and cuticular plates. AB - Auditory hair cells that survive mechanical injury in culture begin their recovery by reforming the kinocilium. This study is based on cultures of the organ of Corti of newborn mice and two control animals. The axonemal patterns were examined in 165 kinocilia in cross-section. In the immature and regenerating kinocilium, one of the normally peripheral doublets is frequently located inward, forming the modified 8 + 1 (double) form; the distribution of the remaining microtubules is irregular. As the cell matures, the 9 + 0 form predominates. Overall, 34-61% of auditory kinocilia consist of 9 + 0 microtubules. The 9 + 2 (single) form, previously thought to characterize the organelle, occurs only in about 3-14%, whereas the remaining population comprises the modified 8 + 1 (double) form. Normally, the kinocilium lasts only about 10 postnatal days; however, post-traumatic hair cells reform their kinocilia regardless of age. Concomitant with the regrowth of the kinocilium, the basal body and its cilium take a central location in the cuticular plate, stereocilia regrow, and the cytoplasmic area adjacent to the basal body displays pericentriolar fibrous densities, growth vesicles, and microtubules, all surrounded by actin filaments. Pericentriolar bodies nucleate microtubules. Involvement of microtubules is seen in the alignment of actin filaments and in the formation of the filamentous matrix of the cuticular plate. We propose that reformation of the kinocilium in recovering post-traumatic hair cells indicates the possible role of its basal body in the morphogenesis and differentiation of cuticular plates and stereocilia. PMID- 7500121 TI - Expression of sodium channel alpha- and beta-subunits in the nervous system of the myelin-deficient rat. AB - Using subtype-specific riboprobes and a non-isotope in situ hybridization technique, the pattern of expression of the mRNAs for voltage dependent sodium channel alpha-subunits I, II, III and NaG, and the beta 1-subunit were compared in myelin-deficient rats and unaffected male littermates. Tissues examined included the hippocampus, cerebellum, spinal cord and dorsal root ganglia. Previous studies have demonstrated that the expression of sodium channel alpha- and beta 1-subunits follows a distinct temporal and spatial pattern during development, characterized in part by greater expression of alpha-subunit III and its mRNA during development than in the adult. We examined animals of 20-22 days of age, a time when, according to earlier reports, the unaffected animals should nearly have reached an adult expression pattern. Normal male littermates were indeed found to express a sodium channel subunit mRNA pattern generally consistent with previous reports on adult rats. Myelin-deficient animals exhibited an expression pattern identical to the unaffected littermates, indicating that myelination is not required for the progression from the embryonic to the adult expression pattern of sodium channel subunits. PMID- 7500123 TI - Synapses between cones and diffuse bipolar cells of a primate retina. AB - The photoreceptor synapses of three representative cells of the six types of diffuse bipolar cell of the rhesus macaque monkey's retina are described at 3.5 4.0 mm eccentricity. Bipolar cell DB3 was found to be postsynaptic to 11 cones at 155 basal synapses; about 70% of these were triad associated. Bipolar cell DB4 as postsynaptic to eight cones at 52 ribbon synapses; in addition it was found also to make an average of two or three basal (non-ribbon) synapses per cone (total 23). The DB5 bipolar cell type had 57 invaginating synapses with seven cones. It too had basal synapses, but only two with each of three cones. The diffuse invaginating bipolar cell described by Mariani (1981) is identified as a member of the DB5 category. Dendrites of cone bipolar cell types which have axons ending in the a-layer of the inner plexiform layer make only basal synapses with the cone pedicle. Those so far investigated are the flat midget bipolar cell and the DB2 and DB3 flat diffuse bipolar cells. All bipolar cells whose axons terminate in the b-layer of the inner plexiform layer are postsynaptic at the ribbon synapses of the cone pedicles. They now appear to fall into two groups. Those whose dendrites are exclusively postsynaptic at the ribbons; these are the blue cone and invaginating midget bipolar cells. And the diffuse bipolar cell DB4, that has both ribbon and basal synapses in a ratio of about 2.3:1. It is uncertain into which category cell DB5 should be placed; its basal synapses are so few the cell could be anomalous. It now seems that at least one primate bipolar cell type may be like those of other vertebrates in having, as defined ultrastructurally, two different kinds of synaptic connection with its cones. The results are discussed in the context of a brief review of the photoreceptor synapses of other mammalian bipolar cells. PMID- 7500122 TI - Schwann cells degrade myelin and proliferate in the absence of macrophages: evidence from in vitro studies of Wallerian degeneration. AB - Interruption of axonal continuity in peripheral nerve trunks leads to axonal and myelin breakdown and removal distal to the injury site, a process known as Wallerian degeneration. Clearance of axonal and myelin debris has been attributed to the cooperative actions of two cell types, the indigenous Schwann cells and macrophages recruited to the regions of tissue damage. Recent work in this area has suggested a limited role for Schwann cells in myelin degradation and has emphasized the role of macrophages, not only in myelin clearance but also in the stimulation of Schwann cell proliferation which also occurs during Wallerian degeneration. In this report, we demonstrate that rat Schwann cells are capable of substantial myelin degradation unaided by macrophages. Observations were made following excision of neuronal somata from well-myelinated rat dorsal root ganglion neuron/Schwann cell co-cultures. The various stages of myelin breakdown were observed by phase microscopy, Sudan black staining, or electron microscopy. The time course for breakdown of individual myelin internodes varied from 2 to 10 days after injury and was to some extent dependent upon the original internodal length. Additionally, we show that most Schwann cells involved in Wallerian degeneration in the absence of macrophages undergo cell division following degradation of myelin into granules visible by light microscopy. The co-cultures employed were essentially free of macrophages as assessed by immunostaining for the OX42, ED2, and ED1 macrophage markers. No macrophages were detected by light or electron microscopy in the vicinity of the identified Schwann cells and furthermore, macrophages/monocytes were rarely observed in uninjured co-cultures as assessed by fluorochrome-conjugated acetylated LDL labelling. These results provide evidence in support of the ability of Schwann cells to carry out degradation of short myelin segments and to proliferate without macrophage assistance during Wallerian degeneration in vitro. PMID- 7500124 TI - Adhesion molecule expression on murine cerebral endothelium following the injection of a proinflammagen or during acute neuronal degeneration. AB - The acute inflammatory response in the murine CNS is different from that observed in other tissues. Few polymorphonuclear leukocytes are recruited to the brain parenchyma and there is a delay in the recruitment of monocytes. Leukocyte recruitment to sites of inflammation is dependent on adhesion molecules expressed on the endothelium. The atypical kinetics of leukocyte recruitment to the CNS may be the result of deficient or delayed adhesion molecule expression on the cerebral endothelium. Using immunohistochemistry, the present study demonstrates that following the intracranial injection of a proinflammagen, lipopolysaccharide, or following acute neuronal degeneration elicited with kainic acid, the adhesion molecules ICAM-1 and VCAM were readily upregulated on cerebral endothelium in a time course comparable with that demonstrated on non-CNS endothelium. Both molecules were expressed on vessels, irrespective of their size, at 24 h after kainic acid or 6 h after lipopolysaccharide injection but leukocyte recruitment was negligible. The expression of ICAM-1 was demonstrated not only on endothelium but also on microglia especially in response to nerve terminal degeneration. PECAM was constitutively expressed at high levels on cerebral endothelium and did not change during brain injury. However, PECAM was induced on astrocytes after lipopolysaccharide injection or during acute neuronal degeneration, the latter providing a particularly strong stimulus. This study indicates that the expression of these adhesion molecules on CNS endothelium is neither deficient or delayed and that they are unlikely to be limiting factors in leukocyte recruitment to the CNS. PMID- 7500125 TI - A giant nerve net with multi-effector synapses underlying epithelial adhesive strips in the mouth of Beroe (Ctenophora). AB - We present ultrastructural evidence for the first known example of a giant nerve net in the phylum Ctenophora. The giant fibre system in Beroe underlies paired strips of adherent epithelial cells that run inside the lips. Interlocking actin lined cell junctions between opposing adhesive strips keep Beroe's large mouth closed while the ctenophore searches for prey. The giant neurons, up to 6-8 microns in diameter, form a continuous lattice-like plexus rich in vesicles, microtubules, and 'presynaptic triads'. A novel feature is that individual giant axons make synaptic contacts with more than one type of effector, i.e. longitudinal muscle fibres and epithelial adhesive cells. Contact of prey with sensory receptors on the lips of Beroe induces rapid disappearance of the actin lined adhesive cell junctions, and muscular opening of the mouth to ingest prey. Electron microscopy of food-opened mouths shows local thickening of longitudinal muscles and widening of the basal ends of epithelial cells in the adhesive strip, correlated with retraction of the adhesive epithelium into the mesoglea. Addition of 1% Triton X-100 to formaldehyde fixative in the absence of prey also elicits regional thickening of longitudinal muscles at the location of the adhesive strips (visualized by rhodamine-phalloidin staining). The giant neuron system may serve as a final common pathway to rapidly signal disassembly of actin-based junctions between adhesive cells as well as contractions of longitudinal muscles underlying the adhesive strips, thereby enabling Beroe to open its mouth rapidly to engulf prey. PMID- 7500126 TI - Long-term potentiation in an isolated peripheral nerve-spinal cord preparation. AB - 1. Long-lasting increases in synaptic efficacy following repetitive stimulation have been demonstrated at several sites in the CNS, where they are collectively termed long-term potentiation (LTP). LTP is of interest with respect to its presumptive relationship to learning and memory in hippocampus. In the spinal cord in vivo, an LTP-like phenomenon is thought to underlie the allodynia and hyperalgesia that follows some peripheral injuries. 2. We investigated the capacity of the isolated neonatal rat spinal cord to sustain a long-lasting increase in a nociceptive-related slow ventral root potential (sVRP) recorded from a lumbar root after a tetanic train of stimuli to the peripheral cutaneous saphenous nerve. Stimuli were delivered at a low constant (0.02 s-1) frequency during a 30-min control period. A tetanic stimulus train (10 s-1 for 60 s) was then given followed by a resumption of low (0.02 s-1) frequency stimulation. Potentiation was defined as an increase in sVRP area > 2 SD above control mean. 3. Twenty of 20 preparations showed immediate posttetanic potentiation. In 13 of the 20, potentiation was maintained for > or = 1 h after the tetanic stimulus train. 4. Potentiation was dependent on activation of C fibers during the inducing train; stimuli below C-fiber threshold, activating only A fibers, were ineffective. Potentiation was selectively expressed by a long-latency component of the sVRP elicited by stimuli at a strength that evoked both A- and C-fiber responses in the nerve.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7500127 TI - Pruning of rat cortical taste neurons by an artificial neural network model. AB - 1. Taste qualities are believed to be coded in the activity of ensembles of taste neurons. However, it is not clear whether all neurons are equally responsible for coding. To clarify the point, the relative contribution of each taste neuron to coding needs to be assessed. 2. We constructed simple three-layer neural networks with input units representing cortical taste neurons of the rat. The networks were trained by the back-propagation learning algorithm to classify the neural response patterns to the basic taste stimuli (sucrose, HCl, quinine hydrochloride, and NaCl). The networks had four output units representing the basic taste qualities, the values of which provide a measure for similarity of test stimuli (salts, tartaric acid, and umami substances) to the basic taste stimuli. 3. Trained networks discriminated the response patterns to the test stimuli in a plausible manner in light of previous physiological and psychological experiments. Profiles of output values of the networks paralleled those of across-neuron correlations with respect to the highest or second-highest values in the profiles. 4. We evaluated relative contributions of input units to the taste discrimination of the network by examining their significance Sj, which is defined as the sum of the absolute values of the connection weights from the jth input unit to the hidden layer. When the input units with weaker connection weights (e.g., 15 of 39 input units) were "pruned" from the trained network, the ability of the network to discriminate the basic taste qualities as well as other test stimuli was not greatly affected. On the other hand, the taste discrimination of the network progressively deteriorated much more rapidly with pruning of input units with stronger connection weights. 5. These results suggest that cortical taste neurons differentially contribute to the coding of taste qualities. The pruning technique may enable the evaluation of a given taste neuron in terms of its relative contribution to the coding, with Sj providing a quantitative measure for such evaluation. PMID- 7500128 TI - Development of spontaneous activity and mechanosensitivity in axotomized afferent nerve fibers during the first hours after nerve transection in rats. AB - 1. Spontaneous activity and ectopic mechanical excitability of axotomized unmyelinated and myelinated fibers in the sural nerve were examined in anesthetized rats. The analysis was performed within 30 h after the nerve lesion using single-fiber recordings that were performed proximal to the severed nerve end. 2. Among all unmyelinated fibers tested (n = 865), 4-8% exhibited persistent spontaneous activity of low and irregular frequency. The percentage of spontaneously active C fibers did not change significantly during the first 30 h. Only 6 of 796 A fibers had spontaneous activity. 3. Mechanical stimulation of the cut nerve end excited 5-8% of all C fibers under investigation. No development with time could be detected in the frequency of mechanically excitable C fibers. In contrast, beginning 6 h after nerve transection, the number of mechanically excitable A fibers rose with time, reaching 27% after 22-30 h. 4. Among the A fibers (C fibers) that exhibited mechanical excitability or spontaneous activity, only 4% (25%) had both properties, whereas 96% (75%) were either mechanosensitive or spontaneously active. 5. With time after the nerve lesion, the mean discharge rate of all spontaneously discharging C fibers decreased significantly from 49 imp/min (0.5-9 h after nerve lesion) to 11 imp/min after 22-30 h. The mean discharge rate of C fibers exhibiting solely spontaneous activity and those C fibers that were additionally mechanosensitive did not differ significantly.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7500129 TI - Somatostatin modulates high-voltage-activated Ca2+ channels in freshly dissociated rat hippocampal neurons. AB - 1. The effects of somatostatin (SS) on the low-voltage-activated and high-voltage activated (HVA) Ca2+ channels in pyramidal neurons acutely dissociated from the hippocampal CA1 region of 2- to 3-wk-old rats were investigated in a nystatin perforated-patch recording configuration under voltage-clamp conditions. 2. SS had no effect on the low-voltage-activated Ca2+ channel but did inhibit the HVA Ca2+ channel in a concentration-, time-, and voltage-dependent manner. 3. SS showed the activation phase of Ba2+ current (IBa) passing through HVA Ca2+ channels, and the maximum inhibition was 28% of the total current amplitude measured 10 ms after the current activation. The inhibitory effect was eliminated by applying larger depolarizing prepulses. Pretreatment with pertussis toxin (PTX) completely blocked the effect of SS on HVA IBa, suggesting the contribution of PTX-sensitive Gi/Go proteins to the SS-induced inhibition. 4. The applications of forskolin, 8-Br-cAMP, dibutyryl-guanosine 3'5'-cyclic monophosphate, staurosporine, and 1-(5-isoquinolinylsulphonyl)-2-methylpiperazine did not affect either the control HVA IBa or the SS-induced inhibition of HVA IBa. 5. Pretreatment with protein kinase C (PKC) activators had no significant effect on HVA IBa but did remove the inhibition of HVA IBa by SS. 6. Omega-Conotoxin-GVIA, omega-agatoxin-IVA, nicardipine, and omega-conotoxin-MVIIC blocked HVA IBa by 27, 13, 38, and 9% of the total HVA current, respectively, which suggested the existence of N-, P-, L-, and Q-type HVA Ca2+ channels in the hippocampal CA1 pyramidal neurons.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7500130 TI - Modulation of muscle responses evoked by transcranial magnetic stimulation during the acquisition of new fine motor skills. AB - 1. We used transcranial magnetic stimulation (TMS) to study the role of plastic changes of the human motor system in the acquisition of new fine motor skills. We mapped the cortical motor areas targeting the contralateral long finger flexor and extensor muscles in subjects learning a one-handed, five-finger exercise on the piano. In a second experiment, we studied the different effects of mental and physical practice of the same five-finger exercise on the modulation of the cortical motor areas targeting muscles involved in the task. 2. Over the course of 5 days, as subjects learned the one-handed, five-finger exercise through daily 2-h manual practice sessions, the cortical motor areas targeting the long finger flexor and extensor muscles enlarged, and their activation threshold decreased. Such changes were limited to the cortical representation of the hand used in the exercise. No changes of cortical motor outputs occurred in control subjects who underwent daily TMS mapping but did not practice on the piano at all (control group 1). 3. We studied the effect of increased hand use without specific skill learning in subjects who played the piano at will for 2 h each day using only the right hand but who were not taught the five-finger exercise (control group 2) and who did not practice any specific task. In these control subjects, the changes in cortical motor outputs were similar but significantly less prominent than in those occurring in the test subjects, who learned the new skill.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7500131 TI - Intracellular Ca2+ stores can account for the time course of LTP induction: a model of Ca2+ dynamics in dendritic spines. AB - 1. A model of Ca2+ dynamics in spines of CA1 hippocampal neurons is presented. In contrast to traditional models, which concentrate on the effects of Ca2+ influx, diffusion, buffering, and extrusion, we also consider the additional effect of intracellular Ca2+ stores. 2. It is shown that traditional models without Ca2+ stores cannot account for the time course of long-term potentiation (LTP) induction as found in recent experiments. Experimental data suggest that the intracellular Ca2+ concentration should be elevated for up to 2 s, whereas the Ca2+ concentration in standard models of Ca2+ dynamics decays much faster. 3. When intracellular Ca2+ stores are taken into account, a much slower decay is found. In particular, a model simulation with a stimulation paradigm consisting of two bursts of four impulses at 100 Hz each and variable interburst intervals can reproduce experimental results found for primed or theta-burst stimulation. 4. In our model, Ca2+ release from the store has a nonlinear, bell-shaped dependence on the intracellular Ca2+ concentration, similar to the one found for inositoltrisphosphate and ryanodine receptors. These receptors are known to control calcium release from intracellular stores. 5. Our model suggests an important role of intracellular calcium stores in the induction of LTP. The stores serve as a long-term calcium source that can sustain an intracellular Ca2+ concentration above the resting level for 1-2 s. PMID- 7500132 TI - Long-term potentiation of glycinergic inhibitory synaptic transmission. AB - 1. Tetanizing protocols were used to test whether glycinergic inhibition undergoes long-term plasticity in vivo. For this purpose we studied the inhibition evoked disynaptically in the teleost Mauthner (M) cell by stimulation of the posterior branch of the contralateral VIIIth nerve. The advantage of this experimental design is that the inhibition, which is mediated by identified second-order commissural interneurons, is not contaminated by parallel excitation. 2. The VIIIth-nerve-evoked inhibitory postsynaptic potentials (IPSPs), which are generated at the level of the soma, are depolarizing in Cl(-) loaded M cells. After VIIIth nerve tetanization, these IPSPs exhibited potentiation lasting > 30 min in 23 of 31 cells. The maximum enhancement measured 5-10 min after the onset of the tetanization averaged 100 +/- 19% (mean +/- SE). In contrast, the non-"tetanized" collateral IPSP induced by antidromic stimulation of the M axon did not increase significantly suggesting synaptic specificity of the potentiation. 3. Single-electrode voltage-clamp studies of Cl( )-loaded M cells indicated that this plasticity is due to an increased synaptic conductance that occurs without obvious modifications of the kinetics or voltage dependence of the inhibitory postsynaptic currents. 4. The synaptic conductance and its changes during potentiation were quantified by measuring the inhibitory shunt of the antidromic spike while recording with potassium-acetate-filled electrodes. For this purpose the ratio, r', of the inhibitory to resting membrane conductances, was calculated using the expression (V/V')--1, where V and V' are the amplitudes of the control and the test antidromic spikes, respectively. This ratio was called fractional conductance. Measured at the peak of the expected VIIIth-nerve-evoked IPSP, r' increased by 114 +/- 18% (n = 46). Again the collateral inhibitory conductance was not modified. 5. Because there are two synapses in the inhibitory pathway, it became important to determine whether modifications of the second-order inhibitory junctions contribute to the overall potentiation. Several experimental procedures were used for this purpose. 6. The input-output relationship at the inhibitory synapses was determined by comparing the size of the presynaptic volley and r'. The former was recorded intra- or extracellularly as a monophasic positive potential, the so-called extrinsic hyperpolarizing potential, which increases in parallel with the strength of VIIIth nerve stimulation. In 12 experiments where the presynaptic volley was unaffected by the tetanization, suggesting lack of involvement of the first relay, r' nevertheless increased in amplitude by 79 +/- 14%.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 7500133 TI - Reexamination of the effects of MCPG on hippocampal LTP, LTD, and depotentiation. AB - 1. We examined the effects of the metabotropic glutamate receptor (mGluR) antagonist alpha-methyl-4-carboxyphenylglycine (MCPG) on the induction of long term potentiation (LTP) long-term depression (LTD), and depotentiation in CA1 hippocampal neurons using extracellular recording techniques. 2. MCPG (500 microM) strongly antagonized the presynaptic inhibitory action of the mGluR agonist 1-aminocyclopentane-(1S,3R)-dicarboxylic acid yet failed to block LTP induced with either tetanic stimulation (100 Hz, 1 s) or theta-burst stimulation. 3. To test the possibility that our failure to block LTP was due to prior activation of a "molecular switch" that in its "on" state obviates the need for mGluR activation to generate LTP, we gave repeated periods of prolonged low frequency stimulation (LFS; 1 Hz, 10 min), a manipulation reported to turn the switch "off." Although this stimulation saturated LTD, subsequent application of MCPG still failed to block LTP. 4. MCPG did not block LFS-induced depotentiation in older slices (4-6 wk) or LFS-induced LTD in older, young (11-18 days), or neonatal (3-7 days) slices. 5. These results demonstrate that MCPG-sensitive mGluRs are not necessary for the induction of LTP, LTD, or depotentiation in hippocampal CA1 pyramidal cells. The possibility remains, however, that their activation may modify the threshold for the induction of these long-term plastic changes. PMID- 7500134 TI - Responses of cells in the tail of the caudate nucleus during visual discrimination learning. AB - 1. The tail of the caudate nucleus and adjacent ventral putamen (ventrocaudal neostriatum) are major projection sites of the extrastriate visual cortex. Visual information is then relayed, directly or indirectly, to a variety of structures with motor functions. To test for a role of the ventrocaudal neostriatum in stimulus-response association learning, or habit formation, neuronal responses were recorded while monkeys performed a visual discrimination task. Additional data were collected from cells in cortical area TF, which serve as a comparison and control for the caudate data. 2. Two monkeys were trained to perform an asymmetrically reinforced go-no go visual discrimination. The stimuli were complex colored patterns, randomly assigned to be either positive or negative. The monkey was rewarded with juice for releasing a bar when a positive stimulus was presented, whereas a negative stimulus signaled that no reward was available and that the monkey should withhold its response. Neuronal responses were recorded both while the monkey performed the task with previously learned stimuli and while it learned the task with new stimuli. In some cases, responses were recorded during reversal learning. 3. There was no evidence that cells in the ventrocaudal neostriatum were influenced by the reward contingencies of the task. Cells did not fire preferentially to the onset of either positive or negative stimuli; neither did cells fire in response to the reward itself or in association with the motor response of the monkey. Only visual responses were apparent. 4. The visual properties of cells in these structures resembled those of cells in some of the cortical areas projecting to them. Most cells responded selectively to different visual stimuli. The degree of stimulus selectivity was assessed with discriminant analysis and was found to be quantitatively similar to that of inferior temporal cells tested with similar stimuli. Likewise, like inferior temporal cells, many cells in the ventrocaudal neostriatum had large, bilateral receptive fields. Some cells had "doughnut"-shaped receptive fields, with stronger responses in the periphery of both visual fields than at the fovea, similar to the fields of some cells in the superior temporal polysensory area. Although the absence of task-specific responses argues that ventrocaudal neostriatal cells are not themselves the mediators of visual learning in the task employed, their cortical-like visual properties suggest that they might relay visual information important for visuomotor plasticity in other structures. (ABSTRACT TRUNCATED AT 400 WORDS) PMID- 7500135 TI - Sensorimotor-correlated discharge recorded from ensembles of cerebellar Purkinje cells varies across the estrous cycle of the rat. AB - 1. In the present study, locomotor-correlated activity of cerebellar Purkinje cells, recorded using arrays of microwires chronically implanted in adult female rats, was examined across estrous-cycle-associated fluctuations in endogenous sex steroids. Ongoing studies from this laboratory have shown that systemic and local administration of the sex steroid 17 beta-estradiol (E2) augments excitatory responses of cerebellar Purkinje cells to iontophoretically applied glutamate, recorded in vivo from anesthetized female rats. In addition, this steroid potentiated discharge correlated with limb movement. For the present study, extracellular single-unit activity was recorded from as many as 5-11 Purkinje cells simultaneously during treadmill locomotion paradigms. Motor modulation of activity was recorded across three to five consecutive estrous cycles from behaviorally identified cohorts of neurons to test the hypothesis that fluctuations in endogenous sex steroids alter motor modulation of Purkinje cell discharge. 2. Locomotor-associated discharge correlated with treadmill locomotion was increased by a mean of 47% on proestrus, when E2 levels are elevated, relative to diestrus 1. These changes in discharge rate during treadmill locomotion were of significantly greater magnitude than corresponding cyclic alterations in discharge during stationary periods. 3. Correlations with the circadian cycle were also significant, because peak levels of locomotor associated discharge on the night of behavioral estrus, following elevations in circulating E2, were on average 67% greater than corresponding discharge recorded during the light (proestrus). 4. Alterations in the step cycle were also observed across the estrous cycle: significant decreases in the duration of the flexion phase (by 265 ms, P < 0.05) were noted on estrus compared with diestrus. 5. When recorded on estrus, Purkinje cell discharge correlated with the stance or flexion phase of the step cycle was greater in magnitude and preceded the event by an average of 130 ms, compared with values determined on diestrus. 6. On estrus, responses of Purkinje neurons to iontophoretically applied quisqualate were enhanced fourfold after administration of exogenous E2, assessed in urethan anesthetized female rats. 7. In addition, systemic administration of E2 (30 ng iv) potentiated responses of cerebellar Purkinje cells to electrical stimulation of the forepaw by an average of 150%, recorded in anesthetized female rats. 8. These results are consistent with the hypothesis that elevations in circulating E2 are associated with enhanced discharge of cerebellar Purkinje cells in response to pharmacological or electrical stimuli or associated with locomotor behavior. PMID- 7500136 TI - Effects of inhibitory amino acid antagonists on reciprocal inhibitory interactions during rhythmic motor activity in the in vitro neonatal rat spinal cord. AB - 1. The role of inhibitory amino acid transmission in the coordination and generation of rhythmic motor activity was examined with the use of an in vitro neonatal rat spinal cord preparation. Before adding gamma-aminobutyric acid (GABA) or glycine receptor agonists and antagonists, rhythmic motor activity was induced by bath application of acetylcholine (ACh), N-methyl-D,L-aspartate (NMA), or serotonin (5-HT) while monitoring bilateral ankle flexor and extensor electroneurograms (ENGs). The timing of rhythmic flexor and extensor discharge was consistent with that seen during overground locomotion in 27% of 84 bath applications of these substances (n = 65 preparations). 2. Subsequent addition of the GABAA receptor agonist muscimol, the GABAB receptor agonist baclofen, or glycine, abolished rhythmic activity in 95% of the tested applications. 3. GABAB receptor blockade did not disrupt alternating patterns of ENG discharge. However, addition of the GABAA receptor antagonist bicuculline, or the glycine receptor antagonist strychnine, transformed alternating flexor-extensor and left-right activity into patterns characterized by bilaterally synchronous rhythmic activation of all hindlimb ENGs. The onset of individual ENG bursts was more abrupt following bicuculline or strychnine. Strychnine also synchronized high frequency (4-8 Hz) packets of rhythmic discharge within ENG bursts. 4. Some preparations developed synchronous, but unstable, rhythmic activity in the presence of bicuculline or strychnine alone. However, NMA, 5-HT, or ACh was usually required in addition to these antagonists to promote sustained rhythmic activity.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7500137 TI - Activation of metabotropic glutamate receptors increase the frequency of spontaneous GABAergic currents through protein kinase A in neonatal rat hippocampal neurons. AB - 1. The tight-seal whole cell recording technique was used to study the effects of the metabotropic glutamate receptor (mGluR) agonist, trans-1-aminocyclopentane 1,3-dicarboxylic acid (t-ACPD) on spontaneous gamma-aminobutyric acid (GABA) mediated synaptic currents in neonatal rat CA1 hippocampal neurons in slices obtained from postnatal (P) days P6-P12. 2. Bath application of t-ACPD (3-30 microM), in the presence of kynurenic acid, induced a concentration-dependent increase in frequency but not in amplitude of spontaneous GABAergic currents. The mean frequency ratio (t-ACPD 10 microM over control) was 2.6 +/- 1 (mean +/- SD), whereas the mean amplitude ratio was 1.1 +/- 0.3. 3. The effect of t-ACPD was partially antagonized by the mGluR antagonist (RS)-alpha-methyl-4 carboxyphenylglycine (MCPG, 1 mM). 4. t-ACPD (10-30 microM) did not modify the frequency of miniature GABAergic synaptic currents recorded in tetrodotoxin (the mean frequency ratio of t-ACPD over control was 0.7 +/- 0.3). 5. Forskolin (30 microM), but not its analogue 1,9 dideoxyforskolin (30 microM), mimicked the effect of t-ACPD. Similar effects were obtained with 3-isobutyl-1-methylxanthine (IBMX, 200 microM). 6. The potentiating effect of t-ACPD on spontaneous GABAergic currents was prevented by Rp-cAMPS (30 microM), a specific antagonist of protein kinase A. This suggests that mGluRs localized at the soma-dendritic level of GABAergic interneurons and positively coupled to cyclic AMP may modulate GABA release during a critical period of postnatal development. PMID- 7500138 TI - Synaptic current kinetics in a solely AMPA-receptor-operated glutamatergic synapse formed by rat retinal ganglion neurons. AB - 1. Postnatal rat retinal ganglion cells (RGCs) can be maintained and identified in dissociated long-term culture. After 4-7 days in vitro they form glutamatergic synapses with other RGCs or putative amacrine cells. Here we intended to characterize the postsynaptic features of these in vitro synapses. 2. Pair patch clamp recordings in the whole cell configuration were performed to study the properties of synaptic glutamate receptors. Immunohistochemically and physiologically identified RGCs were activated by short depolarizing voltage steps. This elicited glutamatergic excitatory postsynaptic currents (EPSCs) in coupled neurons. At room temperature, evoked EPSCs (eEPSCs) had latencies between 3 and 7 ms and amplitudes between 36.4 and 792.6 pA. 3. Postsynaptic neurons were electrotonically compact and therefore well suited for analysis of fast synaptic events. All cells were responsive to exogenous glutamate, alpha-amino-3-hydroxy-5 methyl-4-isoxazolepropionic acid (AMPA) and N-methyl-D-aspartate (NMDA). The current-voltage relationships of AMPA-activated currents were linear, whereas NMDA-induced whole cell currents displayed the typical characteristics including a negative slope conductance in the presence of Mg2+. In contrast to AMPA activated currents, NMDA-activated currents had the usual slow onset and decay. 4. RGCs obviously failed to generate NMDA-receptor-mediated EPSCs, because all postsynaptic cells lacked a slow current component even in the absence of added Mg2+ and in the presence of glycine. Retinal eEPSCs were completely blocked by 6,7-dinitroquinoxaline-2,3-dione (DNQX). 5. eEPSCs as well as spontaneous EPSCs (sEPSCs) were characterized by a very rapid time course. In eEPSCs, 20-80% rise times and time constants of decay (tau DS) were on average 0.64 and 1.96 ms, respectively. eEPSCs were extremely fast, with average rise times of 0.34 ms and tau DS of 1.20 ms. The latter numbers closely correspond to the values obtained for DNQX-sensitive miniature EPSC (mEPSC) in postnatal day 5 rat RGCs in situ. 6. To clarify whether the decay of fast AMPA-receptor-mediated EPSCs of retinal neurons was determined by the onset of glutamate receptor desensitization, we compared the decay of sEPSCs with the decay of the glutamate response of excised out-side-out membrane patches. Glutamate-activated currents were elicited by a rapid superfusion device (time constant of rise = 0.7 ms). The response to 1 mM of glutamate decayed 2 to 4 times more slowly than the sEPSCs. 7. These results suggest that desensitization did not limit the rate of decay of purely AMPA mediated EPSCs in response to ganglion cell activation.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 7500140 TI - Differences in inhibitory synaptic input between layer II-III and layer V neurons of the cat neocortex. AB - 1. The goal of this study was to compare the relative effectiveness of intrinsic inhibitory synaptic inputs in different layers of the cat motor cortex. Postsynaptic potentials (PSPs) were evoked in neurons located in the superficial (layer II-III) or deep layers (layer V) by local extracellular stimulation in vitro. Electrophysiological properties and intracellular filling indicated that the recorded neurons were pyramidal cells. 2. The shape and time course of the evoked PSPs differed. Layer II-III cells showed stereotyped triphasic PSPs consisting of a fast excitatory PSP (fEPSP) and a fast and slow inhibitory PSP (fIPSP and sIPSP, respectively). PSPs in layer V cells, in contrast, were much more variable, mainly depolarizing at resting membrane potential, and lacked a hyperpolarizing IPSP in 84% of neurons tested at rest. 3. Blockade of glutaminergic neurotransmission with 6-cyano-7-nitroquinoxaline-2,3-dione (CNQX) and D-2-amino-5-phosphonovaleric acid (AP5) revealed that fIPSPs could be evoked in all layer V cells by local stimulation of the superficial or deep layers, even in those that showed small or no IPSPs in control perfusate. Small (< 1 mV) isolated sIPSPs were evoked in only one-fifth of layer V cells when the deep layers were stimulated, and in about one-half of the layer V cells when the superficial layers were stimulated. In layer II-III cells, stimulation of the superficial layers always resulted in fIPSP-sIPSP combinations. No IPSPs could be evoked in layer II-III neurons by stimulating the deep layers after glutaminergic blockade. Selective blockade of gamma-aminobutyric acid-A (GABAA) or GABAB receptor-mediated neurotransmission showed that in both cell types fIPSPs were due to GABAA receptor stimulation, whereas sIPSPs were mediated by GABAB receptors. 4. Isolated fIPSPs were recorded in perfusate containing CNQX, AP5, and the GABAB antagonist CGP 35348. The rise and decay times of the fIPSPs in layer II-III cells were significantly longer than those in layer V cells. Rise and decay times normalized for differences in membrane time constant were not significantly different, however, suggesting that the intrinsic membrane properties of the postsynaptic membrane account for the difference in time course of the fIPSPs in these two cell types. 5. Selective blockade of the inward rectifier current Ih with extracellular Cs+ showed that this conductance functions to shorten and attenuate fIPSPs in layer V cells. In contrast, Ih is absent or small in layer II-III cells, and, consequently, Cs+ had little or no effect on the fIPSPs evoked in these cells.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 7500139 TI - Voltage-dependent calcium channels in rat midbrain dopamine neurons: modulation by dopamine and GABAB receptors. AB - 1. Voltage-dependent calcium channels were studied with whole cell voltage-clamp recordings from neurons enzymatically dispersed from the ventral mesencephalon of rat brains (postnatal days 3-10) and identified as dopamine neurons by 5,7 dihydroxytryptamine autofluorescence. 2. Dopamine neurons had large high threshold calcium currents activated by depolarizations positive to -50 mV. Different components of calcium channel current were not readily distinguishable by voltage dependence or kinetics, but pharmacological experiments showed the existence of different channel types. The overall current had significant components blocked by nimodipine (28%), by omega-conotoxin GVIA (22%), and by omega-agatoxin-IVA (omega-Aga-IVA) (37%), and there was a significant amount of current (16%) remaining in saturating concentrations of all three blockers. 3. High-threshold calcium current was reversibly reduced by the gamma-aminobutyric acid-B (GABAB) receptor agonist baclofen and by dopamine and the D2 receptor agonist quinpirole. Inhibition by GABAB or dopamine agonists developed and reversed within seconds. 4. Quinpirole reduced both omega-conotoxin-sensitive and omega-Aga-IVA-sensitive components of calcium current. 5. With physiological ionic conditions, inward calcium currents were outweighed by outward currents, in part through calcium-activated potassium channels activated by omega-conotoxin sensitive and omega-Aga-IVA-sensitive calcium entry. PMID- 7500141 TI - Magnetic and electrical responses of the human brain to texture-defined form and to textons. AB - 1. We searched for a neurophysical correlate of preattentive texture discrimination by recording magnetic and electric evoked responses from the human brain during the first few hundred milliseconds following the presentation of texture-defined (TD) checkerboard form. The only two textons that changed when the TD checkerboard appeared or disappeared were the local orientation and line termination textons. (Textons are conspicuous local features within a texture pattern). 2. Our evidence that the magnetic response to TD form cannot be explained in terms of responses to the two associated textons is as follows: 1) by dissociating the two responses we showed that the magnetic response to TD form is almost entirely independent of the magnetic response to the local orientation texton; 2) a further distinction between the two responses is that their distributions over the head are different; and 3) the magnetic response to TD form differs from the magnetic response to the line termination texton in both distribution over the head and waveform. We conclude that this evidence identifies the existence of a brain response correlate of preattentive texture discrimination. 3. We also recorded brain responses to luminance-defined (LD) checkerboard form. Our grounds for concluding that magnetic brain responses to the onset of checkerboard form are generated by different and independent neural systems for TD and LD form are as follows: 1) magnetic responses to the onset of TD form and LD form had different distributions over the skull, had different waveforms, and depended differently on check size; and 2) the waveform of the response to superimposed TD and LD checks closely approximated the linear sum of responses to TD checks and LD checks alone. 4. One possible explanation for the observed differences between the magnetic and electric evoked responses is that responses to both onset and offset of TD form predominantly involve neurons aligned parallel to the skull, whereas that is not the case for responses to LD form. PMID- 7500142 TI - Bursting and oscillating neurons of the cat basolateral amygdaloid complex in vivo: electrophysiological properties and morphological features. AB - 1. To characterize the physiological properties of lateral and basolateral (BL) amygdaloid neurons, intracellular recordings were performed in barbiturate anesthetized cats. Morphological identification of recorded cells was achieved by intracellular injection of neurobiotin. Two types of physiologically identified projection neurons were distinguished in the BL and lateral nuclei. 2. The first type of neurons prevailed in the BL nucleus (80% of BL cells). Their resting membrane potential (Vm) averaged -66 +/- 4.9 (SE) mV. They generated stereotyped spike doublets or bursts in response to threshold depolarizing pulses. In most cells, depolarizing pulses of higher amplitude elicited spike bursts or doublets at a shorter latency followed by a nonadapting train of single spikes whose frequency rose with the amplitude of the current pulses. However, 15% of BL bursting neurons generated repetitive spike bursts or doublets in response to prolonged depolarizing current pulses. The response of BL bursting neurons to hyperpolarizing current pulses revealed the presence of slow inward rectification in the form of a depolarizing sag, thus suggesting the presence of a hyperpolarization-activated current. 3. The second type of neurons prevailed in the lateral nucleus. Their resting Vm was quite polarized (-74 +/- 2.85 mV) and they generated slow Vm oscillations (2-10 Hz) upon steady depolarization beyond congruent to -62 mV. The frequency of the oscillation increased with the amount of depolarizing current. In the majority of cells, analysis of voltage responses to subthreshold current pulses revealed the presence of voltage- and time dependent rectification in the depolarizing direction. Current pulses that brought the Vm to -65 mV and beyond elicited a voltage response that reached an early peak and then decayed. Increasing the amplitude of the pulse decreased the latency of the early peak until it triggered an action potential. Current-voltage plots demonstrated inward rectification in the depolarizing direction. At the break of hyperpolarizing current pulses applied at depolarized levels, the Vm overshot prepulse values and generated one or more oscillatory cycles. 4. An important proportion of bursting and oscillating neurons (45.8% and 29%, respectively) were physiologically identified as projection neurons by antidromic invasion from the basal forebrain, entorhinal cortex, or perirhinal cortex. The conduction velocity of bursting and oscillating neurons estimated from the latency of antidromic spikes was low (< or = 2.5 m/s). 5. Most bursting and oscillating neurons of the BL nucleus were spiny cells with a pyramidal morphology. Four to eight dendritic trunks emerged from the apex, base, and sides of their triangular soma.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 7500143 TI - Face-sensitive regions in human extrastriate cortex studied by functional MRI. AB - 1. We have previously identified face-selective areas in the mid-fusiform and inferior temporal gyri in electrophysiological recordings made from chronically implanted subdural electrodes in epilepsy patients. In this study, functional magnetic resonance imaging (fMRI) was used to study the anatomic extent of face sensitive brain regions and to assess hemispheric laterality. 2. A time series of 128 gradient echo echoplanar images was acquired while subjects continuously viewed an alternating series of 10 unfamiliar faces followed by 10 equiluminant scrambled faces. Each cycle of this alternating sequence lasted 12 s and each experimental run consisted of 14 cycles. The time series of each voxel was transformed into the frequency domain using Fourier analysis. Activated voxels were defined by significant peaks in their power spectra at the frequency of stimulus alternation and by a 180 degrees phase shift that followed changes in stimulus alternation order. 3. Activated voxels to faces were obtained in the fusiform and inferior temporal gyri in 9 of 12 subjects and were approximately coextensive with previously identified face-selective regions. Nine subjects also showed activation in the left or right middle occipital gyri, or in the superior temporal or lateral occipital sulci. Cortical volumes activated in the left and right hemispheres were not significantly different. Activated voxels to scrambled faces were observed in six subjects at locations mainly in the lingual gyri and collateral sulci, medial to the regions activated by faces. 4. Face stimuli activated portions of the midfusiform and inferior temporal gyri, including adjacent cortex within occipitotemporal sulci.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7500144 TI - Slowly inactivating outward currents in a cuticular mechanoreceptor neuron of the cockroach (Periplaneta americana). AB - 1. Although rapid adaptation is a widespread feature of sensory receptors, its ionic basis has not been clearly established in any touch receptor, because their small sizes have severely restricted the range of experiments tat can be performed. In the cockroach tactile spine, intracellular voltage-clamp recordings are now possible. 2. The basic electrophysiological properties of the cockroach femoral tactile spine neuron were studied using discontinuous (switching) single electrode current- and voltage-clamp recordings. A slowly inactivating voltage sensitive K+ outward current was detected after the major inward currents were blocked with tetrodotoxin. 3. The total outward current activated in < 1 ms at voltages above 0 mV. At moderate depolarizations it did not inactivate, but at higher depolarizations an inactivation time constant of approximately 260 ms was measured. Some recordings also revealed an additional, slower inactivation time constant of approximately 2.5 s. 4. More than half of the voltage-sensitive K+ outward current could be blocked with the Ca2+ channel blockers Co2+ and Cd2+. Tetraethylammonium chloride (TEA) also reduced the amplitude of the outward current to about half of its original amplitude. The actions of both blockers were reversible and probably reflect overlapping blockades of two separate outward currents. 5. The reversal potentials of the currents that remained after block with Co2+ (-91.7 mV) or TEA (-86.8 mV) were both near the K+ equilibrium potential expected for the tactile spine neuron. The voltage dependencies of activation of the Co(2+)- and TEA-resistant currents were both well fitted by Boltzmann distributions, giving values of half maximal activation (V50) equal to 34.5 mV for the Co(2+)-resistant current and -51.3 mV for the TEA-resistant current. 6. Current-clamp recordings revealed that the TEA-sensitive K+ current was the major component of action potential repolarization but that it did not effect the frequency of action potentials evoked by steady depolarization. On the other hand, blockers of Ca(2+)-sensitive K+ currents (Cd2+, Co2+, or charybdotoxin) reduced adaptation and increased the frequency of action potentials significantly but did not effect the duration or amplitude of individual action potentials. PMID- 7500145 TI - Agonist and toxin sensitivities of ACh-evoked currents on neurons expressing multiple nicotinic ACh receptor subunits. AB - 1. We have investigated the pharmacological properties of functional nicotinic acetylcholine receptors (nAChRs) on neonatal rat sympathetic neurons from the superior cervical ganglion (SCG) to learn more about the subunit composition of these receptors. These neurons express five nAChR transcripts: alpha 3, alpha 5, alpha 7, beta 2, and beta 4; this finding suggests that SCG neurons may express several different, physiologically distinct, subtypes of nAChRs. 2. To identify potential subtypes, we have characterized currents evoked by different nicotinic agonists and determined their sensitivity to blockade by alpha-bungarotoxin (alpha-BTX) and by neuronal bungarotoxin (n-BTX). From dose-response curves, we find that the ED50 for both cytisine and dimethylphenylpiperazinium (DMPP) is 20 microM and for ACh is 52 microM. n-BTX blocks the ACh-gated currents rapidly, but the kinetics for n-BTX removal is dependent on the duration of the application: brief applications were quickly reversible, whereas prolonged applications took orders of magnitude longer to reverse. 3. Using fast (ms) agonist application, we observed no rapidly desensitizing currents despite the high levels of alpha 7 in these neurons, nor did we observe any currents that could be blocked by alpha BTX. 4. Using Xenopus oocytes expressing alpha 7 receptors, we show that choline evokes a significant current that is blocked by alpha-BTX. In contrast, choline is much less potent on alpha 3 beta 4 receptors expressed in Xenopus oocytes. Choline can also act as a weak agonist for nAChRs on rat SCG neurons, but its evoked current is not blocked by alpha-BTX. 5. Our results indicate that, when measured at the macroscopic level, most functional nAChRs on SCG neurons behave as a uniform population of receptors, at least with respect to agonist activation and toxin blockade. In comparison with known receptors expressed in heterologous systems, the physiological properties of ACh-evoked currents on SCG neurons are most similar to receptors that have coassembled with both beta 2 and beta 4. PMID- 7500146 TI - Dynamics of neurons in the cat lateral geniculate nucleus: in vivo electrophysiology and computational modeling. AB - 1. We investigated the time domain transformation that thalamocortical relay cells of the cat lateral geniculate nucleus (LGN) perform on their retinal input, and used computational modeling to explore the biophysical properties that determine the dynamics of the LGN relay cells in vivo. 2. We recorded simultaneously the input (S potentials) and output (action potentials) of 50 cat LGN relay cells stimulated by drifting sinusoidal gratings of varying temporal frequency. The temporal modulation transfer functions (TMTFs) of the neurons were derived from these data. The burstiness of the LGN spike trains was also assessed using objective criteria. 3. We found that the form of the TMTF was quite variable among cells, ranging from low-pass to strongly band-pass. The optimal temporal frequency of band-pass neurons was between 2 and 8 Hz. In addition, the TMTF of some cells was nonstationary: their temporal tuning changed with time. 4. The temporal tuning of a cell was directly related to the degree of burstiness of its spike train. Tonically firing relay cells had low-pass TMTFs, whereas the most bursty neurons exhibited the most sharply band-pass transfer functions. This was also true for single cells that altered their temporal tuning: a shift to more band-pass tuning was associated with increased burstiness of the spike train, and vice versa. 5. We constructed a computer simulation of the LGN relay cell. The model was a simplified five-channel version of the thalamocortical neuron model of McCormick and Huguenard. It incorporated the quantitative kinetics of the Ca2+ T channel, as well as the Hodgkin-Huxley Na+ and K+ channels, as the only active membrane currents. To simulate the in vivo dynamics of the relay cell, the input to the model consisted of trains of synaptic potentials, recorded as S potentials in our physiological experiments. 6. When the resting membrane potential of the model neuron was relatively depolarized, the model's TMTF was low-pass, with no bursting evident in the simulated spike train. At hyperpolarized resting membrane potentials, however, the modeled TMTF was band-pass, with frequent burst discharges. Thus the biophysical model reproduced not only the range of dynamics seen in real LGN relay cells, but also the dependence of the overall dynamics on the burstiness of the spike train. However, neither of these phenomena could be simulated without the T channel. Thus the simulations demonstrated that the T-type Ca2+ channel was necessary and sufficient to explain the LGN dynamics observed in physiological experiments.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 7500147 TI - Evidence for common expression mechanisms underlying heterosynaptic and associative long-term depression in the dentate gyrus. AB - 1. The extent to which heterosynaptic and prime-associative stimulation protocols generate different forms of long-term depression (LTD) was assessed in the lateral perforant path synapses terminating on dentate gyrus granule cells in pentobarbital-anesthetized rats. 2. Heterosynaptic LTD was induced in the lateral path by repeated tetanization of the medial path. Prime-associative LTD of the lateral path was induced by alternating high-frequency conditioning trains to the medial path and single shocks to the lateral path at 100-ms intervals, all occurring 10 min after priming stimulation of the lateral path (5 Hz, 80 pulses). 3. Induction of LTD by one administration of the prime-associative protocol was normally greater in magnitude than the LTD induced by the heterosynaptic protocol. Saturation of LTD by repeated delivery of the prime-associative protocol completely occluded the subsequent induction of LTD by the heterosynaptic protocol. Saturation of LTD by repeated delivery of the heterosynaptic protocol produced an 80% occlusion of the LTD generated by the prime-associative protocol. 4. These data support the hypothesis that activity dependent (associative) and activity-independent (heterosynaptic) LTD involve overlapping expression mechanisms, despite having demonstrably different induction mechanisms. PMID- 7500148 TI - Accumulation of inactivation in a cloned transient K+ channel (AKv1.1a) of Aplysia. AB - 1. Inactivation of a cloned Aplysia K+ channel, AKv1.1a, expressed in Xenopus oocytes was examined by a cell-attached macropatch recording. A fast macroscopic inactivation (the time constant for decay was in the range of 20-40 ms) in response to a depolarizing command pulse was insensitive to the concentration of external K+ (2-100 mM KCl). 2. By contrast, recovery from inactivation was extremely slow and dependent on external K+. When the concentration of external KCl was 2-3 mM, a patched membrane had to be held at hyperpolarized potential for > 40 s for a full recovery. The recovery was greatly accelerated if external K+ concentration was increased. A tail current following a command pulse long enough to inactivate most of the channels showed a marked rising phase. 3. A consequence of the slow recovery from inactivation was that AKv1.1a showed a marked accumulation of the inactivation following repetitive pulses, even at low frequency (< 0.1 Hz). When two depolarizing pulses were applied at a short interval, the current during a second pulse was smaller than the current at the end of the preceding pulse. This is a phenomenon called "cumulative inactivation." The onset and the extent of cumulative inactivation of AKv1.1a were voltage dependent but relatively insensitive to external K+ concentration. An amino terminal deletion mutant of AKv1.1a that lacks the fast N-type inactivation did not show cumulative inactivation. 4. These results suggest that the inactivation gating by the amino terminal region of AKv1.1a has a similarity to open-channel blockade, and that the cumulative inactivation can also be dependent on the amino terminal cytoplasmic domain of K+ channels. PMID- 7500149 TI - Processing of kinetically defined boundaries in the cortical motion area MT of the macaque monkey. AB - 1. Electrophysiological recordings of 68 cells in the middle temporal area MT were made in paralyzed and anesthetized macaque monkeys. 2. Testing with our kinetic boundary stimuli always occurred under optimized conditions. To this end, the preferred direction, speed, stimulus position, and stimulus size of each cell were determined by quantitative tests. 3. The orientation selectivity to stationary luminance contrast edges served as a reference by which a response to kinetic boundaries could be compared. We found cells in area MT to be less selective to the orientation of luminance contrast stimuli than to the direction of motion. We confirmed the presence of neurons with preferred orientation aligned with their preferred direction. 4. The responses to kinetic edges defined by motion vectors moving in opposite directions, kinetic gratings with motion vectors in opposite directions, kinetic edges containing coherent motion and a stationary complementary field or coherent motion and a complementary field containing visual dynamic noise were compared. Kinetic boundaries were generated so that the motion vectors moved either parallel or orthogonal to the orientation of the discontinuity. For a cell to be considered as responding to the orientation of a kinetic boundary, it had to exhibit the same preferred orientation when the local motion vectors changed from parallel to orthogonal to the orientation of the kinetic boundary. 5. All cells in area MT changed their preferred orientation by 90 degrees when the coherent motion vectors changed from moving parallel to moving orthogonal to the boundary. This was the case independent of the types of kinetic boundary tested. We concluded that cells in area MT appear to respond to the motion vector over their classical receptive field (CRF) only and were unable to code the orientation of the kinetic boundary. 6. In those cells exhibiting an antagonistic surround, we examined the ability of the cell to code the position of a kinetic boundary. None of the cells tested signaled the position of a kinetic boundary. The side preference of the stimulus of the cells changed from left to right as the motion vectors in the stimulus reversed. This indicates that the cells were only selective for the motion vectors present over their CRF. 7. We found that the directional sensitivity of cells in area MT remained unaltered by the presence of additional motion vectors within the CRF. This suggests that cells in area MT extract a specific motion vector from a spatial configuration of vectors.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 7500150 TI - Comparison of the responses to moving texture patterns of simple and complex cells in the cat's area 17. AB - 1. Whether complex (C) cells are the only truly texture-sensitive units in the cat's primary visual cortex remains controversial. In view of the strong physiological significance of having putatively only one class of cells sensitive to visual noise in the striate cortex, we reinvestigated this issue. Sensitivities of simple (S) and C cells to noise were quantitatively studied and compared in order to clearly document the response properties of cells in the striate cortex to visual noise and to establish whether one can unequivocally segregate S from C cells on the basis of those specific properties. 2. Receptive fields were stimulated with all relevant stimuli, i.e., drifting sine-wave gratings, electronically generated noise pattern of 256 x 256 elements (ratio 1:1 of dark and light elements), and flashing and moving bars (both bright and dark). 3. A total of 60 S cells out of 85 (70.6%) and 90 C cells out of 101 (81.8%) responded to the motion of visual noise. Responses of most C cells were sustained, i.e., their discharge rate was maintained at a constant level throughout presentation of the stimulus. On the other hand, responses of the majority of S cells were characterized by several bursts of discharges. On average, optimal firing rates were greater for gratings than for noise. 4. For practically all cells, responses to noise varied as a function of direction of motion. The mean direction bandwidths were, respectively, 43 +/- 24 degrees and 48 +/- 23 degrees (mean +/- SD) for S and C cells. In both groups, neurons were more broadly tuned for the direction of noise than that of gratings (t-test, P < 0.001). We rarely observed bimodal tuning curves for noise, with each peak lying on either side of the orientation curve. These results could be expected if one considers texture stimuli not in the space domain (as dot patterns) but in the frequency domain, i.e., patterns containing all spatial frequencies and orientations. 5. In general, the direction indexes of S and C cells were similar whether they were stimulated by drifting noise or gratings. S cells had a slight tendency to be more direction selective for noise than for gratings. 6. For all S and C cells tested, responses to noise varied as a function of drift velocity. The mean optimal velocity was 12.9 and 10.2 degrees/s for S and C cells, respectively (t-test, P > 0.05). Most cells were band-pass with mean bandwidths of 2.2 and 2.7 octaves for S and C cells, respectively.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 7500151 TI - Dopaminergic transmission between identified neurons from the mollusk, Lymnaea stagnalis. AB - 1. Dopaminergic transmission was investigated in the central nervous system (CNS) of the freshwater snail, Lymnaea stagnalis. 2. The giant pedal neuron, designated as right pedal dorsal one (RPeD1), makes chemical, monosynaptic connections with a number of identified follower cells in the CNS. Previous work has shown that RPeD1 is an interneuron and a important component of the Lymnaea respiratory central pattern generator. In this study, the hypothesis that RPeD1 uses dopamine as its neurotransmitter was tested by chromatographic, pharmacological, and electrophysiological methods. Characterization of RPeD1's transmitter pharmacology is essential to clearly understand its role in Lymnaea. 3. Earlier studies demonstrated that the soma of RPeD1 contains dopamine. This was quantitated in the present study by high-performance liquid chromatography (with electrochemical detection) of isolated RPeD1 somata and growth cones, which yielded 0.8 +/- 0.3 and 0.10 +/- 0.08 pmol of dopamine per soma and growth cone, respectively. 4. Bath or pressure application of dopamine to follower cells of RPeD1, in situ, mimicked the effects of RPeD1 stimulation. Dose-response curves were constructed for the excitatory effect of dopamine on follower cells, visceral dorsal two and three (VD2/3) (ED50 = 39 microM; Hill coefficient = 1.03), and the inhibitory effect of dopamine on follower cell, visceral dorsal four (ED50 = 33 microM; Hill coefficient = 0.92). 5. The following dopamine agonists (100 microM) were tested by bath application: 2-amino-6,7-dihydroxy 1,2,3,4-tetrahydronaphthalene (ADTN), apopmorphine, 2-bromo-alpha-ergocryptine, deoxyepinephrine (DE), mesulergine, (-) quinpirole, SKF 38393, and tyramine. Only the general dopamine agonists, ADTN and DE, mimicked RPeD1's effects on its follower cells. 6. When VD2/3 was isolated and plated in vitro, it maintained a depolarizing response to dopamine. This response was reduced by intracellular injection of the G-protein blocker, GDP-beta-S (2 mM in electrode). Similarly, incubation of VD2/3, in vitro for approximately 18 h, with pertussis toxin (PTX; 5 micrograms/ml), the G-protein inactivating exotoxin, also reduced the dopamine response. Injecting GDP or incubating in heat-inactivated PTX did not effect the response. 7. Several dopamine antagonists were used in an attempt to block RPeD1's synapses: chlorpromazine, ergonovine, fluphenazine, haloperidol, 6 hydroxydopamine, SCH 23390, (+/-) sulpiride, and tubocurarine. Only the D-2 dopamine receptor antagonist, (+/-) sulpiride, reversibly blocked synaptic transmission from RPeD1 to its follower cells. Both the (+) and the (-) enantiomer of sulpiride also antagonized synaptic transmission. A dose-inhibition curve for (+/-) sulpiride was constructed (IC50 = 47 microM).(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 7500152 TI - Spindle waves are propagating synchronized oscillations in the ferret LGNd in vitro. AB - 1. The cellular features of propagation of spindle waves and a bicuculline induced slow oscillation through sagittal slices of the ferret dorsal lateral geniculate nucleus (LGNd) maintained in vitro were examined with simultaneous extracellular and intracellular recordings from up to eight sites. Spindle waves typically propagated along the long axis (dorsal-ventral) of the sagittal slice at a speed of 0.3-1.5 mm/s and were synchronized along the line of projection between the perigeniculate nucleus (PGN) and the A, A1, and C laminae. 2. Spindle waves can be initiated with local electrical stimulation or can occur spontaneously in any part of the LGNd/PGN. On initiation of a spindle wave, spindle waves propagate away from the site of initiation. Spindle waves may propagate only locally or may collide with other spindle waves. Collision of spindle waves is associated with synchronization of the two network oscillations, and the spindle waves do not propagate past one another. 3. Repetitive electrical stimulation reveals that spindle wave generation and propagation exhibit a relative refractory period of between 7 and 14 s in vitro. Stimulation at rates of less than the refractory period results in the generation of abbreviated local spindle waves at the stimulation site, but not in the propagation of this spindle wave into adjacent regions of the slice. 4. Local block of non-N-methyl-D aspartate (non-NMDA) excitatory amino acid receptors in the PGN with 6-cyano-7 nitroquinoxaline-2,3-dione (CNQX) prevents the propagation of spindle waves across the point of application, indicating that the excitation of PGN neurons by thalamocortical cells is essential to the propagation of these oscillations. The local antagonism of non-NMDA receptors in the PGN results in the dorsal and ventral aspects of the LGNd slice behaving as independent spindle wave generators, even if before application of CNQX they were not. 5. Activation of a burst discharge in a single PGN neuron can result in the generation of a full spindle wave and the propagation of this spindle wave both dorsally and ventrally through the slice away from the activated PGN neuron. The ability of a burst of action potentials in a PGN neuron to generate a spindle wave is suppressed immediately after the generation of a spindle wave, but slowly returns over a 7- to 14-s period corresponding to the relative spindle wave refractory period.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 7500153 TI - Calcium imaging of motoneuron activity in the en-bloc spinal cord preparation of the neonatal rat. AB - 1. This paper describes the use of calcium imaging to monitor patterns of activity in neonatal rat motoneurons retrogradely labeled with the calcium sensitive dye, calcium green-dextran. 2. Pressure ejection of calcium green dextran into ventral roots and into the surgically peeled ventrolateral funiculi (VLF) at the lumbar cord labeled spinal motoneurons and interneurons. The back labeled motoneurons often formed two or three discrete clusters of cells. 3. Fluorescent changes (10-20%) could be detected in labeled motoneurons after a single antidromic stimulus of the segmental ventral root. These changes progressively increased in amplitude during stimulus trains (1-5 s) at frequencies from 5 to 50 Hz, presumably reflecting a frequency-dependent increase in free intracellular calcium. 4. Stimulation of the ipsilateral VLF at the caudal lumbar level (L6), elicited frequency-dependent, synaptically induced motoneuronal discharge. Frequency-dependent fluorescent changes could be detected in calcium green-labeled motoneurons during the VLF-induced synaptic activation. 5. The spatial spread of synaptic activity among calcium green-labeled clusters of motoneurons could be resolved after dorsal root stimulation. Low-intensity stimulation of the roots produced fluorescence changes restricted to the lateral clusters of motoneurons. With increasing stimulation intensity the fluorescence change increased in the lateral cells and could spread into the medial motoneuronal group. After a single supramaximal stimulus a similar pattern was observed with activity beginning laterally and spreading medially. 6. Substantial changes in fluorescence of calcium green-labeled motoneurons were also observed during motoneuron bursting induced by bath application of the glycine receptor antagonist strychnine or the potassium channel blocker 4-aminopyridine (4-AP). 7. Our results show that membrane-impermeant fluorescent calcium indicators can be used as a tool to study the activity of specific populations of spinal neurons during execution of motor functions in the developing mammalian spinal cord. They also suggest that lateral clusters of motoneurons in the developing spinal cord of the rat are more recruitable or excitable than more medial clusters. Further understanding of these findings requires identification of these clusters. PMID- 7500154 TI - Subthreshold synaptic activation of voltage-gated Ca2+ channels mediates a localized Ca2+ influx into the dendrites of hippocampal pyramidal neurons. AB - 1. Whole cell recordings and high-speed fluorescence imaging were used to investigate the spatial and temporal characteristics of Ca2+ influx during synaptic activity in hippocampal CA1 pyramidal neurons. Brief, subthreshold trains of synaptic potentials elicited by Schaffer collateral stimulation produced transient increases in [Ca2+]i in the apical dendrites near the site of synaptic input. The rises in [Ca2+]i were not due to Ca2+ entry through N-methyl D-aspartate (NMDA)-activated or non-NMDA-activated glutamate channels, but were reduced by low concentrations of Ni2+. Hyperpolarizing prepulses caused an increase in the synaptically evoked Ca2+ transients, whereas strong hyperpolarization during the train prevented the rise in [Ca2+]i. The data suggest that subthreshold synaptic activity can open low-voltage-activated (T type) Ca2+ channels and produce a local increase in intradendritic [Ca2+]. Such local increases in [Ca2+]i may be important for modulating the strength of synaptic connections. PMID- 7500155 TI - Persistent Na+ conductance in medium-sized neostriatal neurons: characterization using infrared videomicroscopy and whole cell patch-clamp recordings. AB - 1. In the present study we investigate the expression of a persistent Na+ conductance (INaP) in identified medium-sized neostriatal neurons. Nomarski optics and infrared videomicroscopy were used for cell visualization and identification in thick slices (350 microns). Current- and voltage-clamp recordings were obtained utilizing whole cell patch-clamp methodology. 2. Application of depolarizing ramp voltage commands from a holding potential of -70 mV induced a slow, noninactivating inward current that occurred before and independent of the rapidly inactivating sodium current that subserves action potential generation. INaP began to activate at potentials less negative than -70 mV and peaked at -34 +/- 1 (SE) mV. Its average peak amplitude was -100 +/- 17 pA. INaP was abolished by tetrodotoxin (TTX, 0.5-1 microM) or an Na(+)-free solution. In contrast, it was not affected by Ca2+ channel blockers. Depolarizing ramp commands also induced tetraethylammonium-sensitive outward currents. 3. Dopamine (DA) (20-100 microM) produced a significant reduction of INaP. 4. These results demonstrate the existence of a TTX-sensitive persistent Na+ conductance in medium-sized neostriatal neurons. This conductance is modulated by DA and could play a role in the generation of rhythmic oscillations and in supporting repetitive firing. PMID- 7500157 TI - Deafferentation increases the intracellular calcium of cochlear nucleus neurons in the embryonic chick. AB - 1. Ratiometric fura-2 imaging was used to measure the intracellular calcium concentration ([Ca2+]i) of neurons in the embryonic avian cochlear nucleus, nucleus magnocellularis (NM), after an in ovo unilateral cochlea removal (deafferentation). 2. The mean [Ca2+]i of NM neurons receiving normal input was 113 nM. 3. Deafferentation increased the mean [Ca2+]i of NM neurons to 247, 311, 339, and 314 nM at 1, 3, 6, and 12 h after cochlear removal, respectively. These values did not differ significantly. 4. The percent frequency distribution of deafferented NM neuron [Ca2+]i shifts away from normative levels toward higher [Ca2+]i at 1 and 3 h after cochlear removal, but shifts back toward normative levels at 6 and 12 h after cochlear removal. 5. This increased [Ca2+]i following cochlear removal temporally coincides with well-characterized changes in NM neurons following activity deprivation. 6. These data suggest that deregulation of [Ca2+]i homeostasis plays a key role in NM neuron degeneration and death following activity deprivation. PMID- 7500156 TI - Evidence for a contribution of ICa to serotonergic modulation of IK,Ca in Hermissenda photoreceptors. AB - 1. The Ca(2+)-dependent K+ current (IK,Ca) contributes to both the plateau phase of light-elicited generator potentials and enhanced excitability of identified type B photoreceptors of Hermissenda detected after classical conditioning. Serotonergic modulation of membrane conductances mimics some of the effects of conditioning. Serotonin (5-HT) reduces the magnitude of IK,Ca and decreases the sustained voltage-activated Ca2+ current (ICa) in type B photoreceptors. We have examined the modulatory role of 5-HT in regulation of IK,Ca by ICa using a Ca2+ ionophore in conjunction with the whole cell patch-clamp technique in isolated photoreceptors. 2. The 40-50% reduction of IK,Ca by 5-HT was voltage independent. Cd2+ blocked ICa and reduced IK,Ca by 70-80%. The remaining 20-30% of IK,Ca may result from Ca2+ release from intracellular stores, because IK,Ca was further reduced to 5-10% as the pipette ethylene glycol-bis(beta-aminoethyl ether) N,N,N',N'-tetraacetic acid (EGTA) was raised from 0.5 to 5 mM. The application of the Ca2+ ionophore A23187, which was designed to produce Ca2+ influx independent of the voltage-activated Ca2+ channels, restored IK,Ca. 3. The application of A23187 reversed the effects of 5-HT and Cd2+ on IK,Ca for experiments lasting 15 20 min. However, for longer time periods (> 25 min), complete restoration of IK,Ca by A23187 was obtained in the presence of Cd2+ but not 5-HT. These results suggest that for 15 to 20 min exposures the reduction of IK,Ca by 5-HT is a consequence of modulation of ICa by 5-HT and not a direct effect of 5-HT on IK,Ca. PMID- 7500158 TI - Predictive human pursuit and "orbital goal" of microstimulated smooth eye movements. AB - 1. Anticipatory saccades in smooth pursuit move the point of gaze from near the moving target to well ahead of it, interrupting accurate smooth pursuit. Their effects on the pursuit process were studied in 22 normal human subjects. We presented horizontal periodic target trajectories of 30 degrees amplitude and 30 degrees/s constant velocity or 0.4 Hz sinusoidal velocity in 40-s trials. Saccades and surrounding smooth eye movement (SEM) segments were marked and classified by computer. 2. Anticipatory saccades were often followed by slowed SEM that tended to intercept the target at the endpoint of its trajectory. This was seen in the distribution of projections of the initial 60 ms of postsaccadic SEM to the time of the trajectory endpoint. Magnitude of this SEM tended to follow a function of the time and location of the endpoint of the anticipatory saccade, decreasing as the anticipatory saccades landed closer to the trajectory endpoint. 3. The time and location of the target trajectory endpoint seemed to be the goal for this SEM. We believe this to demonstrate the predictive use of the period and amplitude of the trajectory in smooth pursuit, apart from the instantaneous velocity match of the target. 4. Gottlieb and coworkers in the frontal eye field and Ron and Robinson in the cerebellum produced SEMs in the monkey by microstimulation. At some sites in both structures, direction and velocity of the SEMs depended on the initial position of the eye in that the elicited SEMs appeared to be converging toward a common point, or "orbital goal", and the SEM velocity diminished as the gaze neared that goal.2+ Both our SEM after anticipatory saccades and microstimulated SEM in the monkey slowed as the initial position was brought closer to the inferred orbital goal. This similarity suggests that the goal-directed SEM sites in the monkey might be part of a mechanism for predictive pursuit. PMID- 7500159 TI - Differences in the brain stem terminations of large- and small-diameter vestibular primary afferents. AB - 1. We visualized the central axons of 32 vestibular afferents from the posterior canal by extracellular application of horseradish peroxidase, reconstructed them in three dimensions, and quantified their morphology. Here we compare the descending limbs of central axons that differ in parent axon diameter. 2. The brain stem distribution of descending limb terminals (collaterals and associated varicosities) varies systematically with parent axon diameter. Large-diameter afferents concentrate their terminals in rostral regions of the medial/descending nuclei. As axon diameter decreases, there is a significant shift of terminal concentration toward the caudal vestibular complex and adjacent brain stem. 3. Rostral and caudal regions of the medial/descending nuclei have different labyrinthine, cerebellar, intrinsic, commissural, and spinal connections; they are believed to play different roles in head movement control. Our data help clarify the functions of large- and small-diameter afferents by showing that they contribute differentially to rostral and caudal vestibular complex. PMID- 7500160 TI - Existence in the nucleus incertus of the cat of horizontal-eye-movement-related neurons projecting to the cerebellar flocculus. AB - 1. Properties of nucleus incertus (NIC) neurons projecting to the cerebellar flocculus were studied in alert cats by using chronic unit and eye movement recording and antidromic activation. Projection of these neurons onto the flocculus was verified with retrograde transport of horseradish peroxidase after injections in the flocculus. 2. Bipolar stimulation electrodes were implanted into the "middle" zone of each flocculus because this zone is known to be involved in the control of horizontal eye movements. The dorsomedial aspect of the pontine tegmentum was explored with microelectrodes during stimulation of both flocculi. The majority of neurons antidromically activated from the flocculus were found in the caudal part of the NIC. 3. Of the 69 neurons activated from the flocculus, 44 were classified as burst-tonic (BT) neurons; 34 discharged in relation with horizontal movements of the eye, 10 in relation with vertical movements. Of the 14 remaining neurons, 6 were not related to eye movements and 8 were classified as burst neurons. The BT neurons of the NIC displayed a great sensitivity to both horizontal eye position and horizontal eye velocity. 4. This study demonstrates the presence of a new group of horizontal eye movement related BT neurons situated in the NIC. The fact that they project to the horizontal floccular zone emphasizes the importance of the functional specialization of the different Purkinje cell zones. PMID- 7500161 TI - Activity in the caudate nucleus of monkey during spatial sequencing. AB - 1. There are indications that the execution of behavioral sequences involves the basal ganglia. In this study we examined the role of the caudate nucleus in the construction, storage, and execution of spatial plans. 2. Two monkeys (Macaca mulatta) were trained to perform sequences of saccades and arm movements. The animals had to remember the order of illumination, variable from one sequence to another, of three fixed spatial targets. After a delay, they had to visually orient toward, and press each target in the same order. Six different sequences were executed on the basis of the order of illumination of the targets. Single cell activity was recorded from the four caudate nuclei of the two monkeys. 3. Neural activity was analyzed in each sequence during 10 different periods: the instruction period in which the targets were illuminated, the three orientation periods toward the different targets, the three postsaccadic periods, and the three periods of target pressing. Statistical comparisons were made to detect differences between the different sequences with respect to activity in each period (sequence specificity). 4. A total of 2,100 neurons were studied, of which 387 were task related. The task-related cells were found in both the head and the body of the caudate nucleus. 5. During central fixation, anticipatory activity (n = 81) preceded onset of specific events. Four groups were considered: 1) neurons (n = 46) anticipating offset of the central fixation point, 2) neurons (n = 7) anticipating the illumination of any target, regardless of its spatial position or order of presentation (rank), 3) neurons (n = 17) anticipating the illumination of the first target, regardless of its spatial position, and 4) neurons (n = 11) anticipating the illumination of a given target, regardless of its rank. 6. Phasic visual responses to target onset were observed in 48 cells. The cells responded primarily to the contralateral and upper targets. In a majority (n = 35), visual responses were modulated by the rank of the target(s). Many cells (n = 20) responded only if the corresponding target was first; other cells responded only if the target was second or if it had complex time relationships with the other targets. 7. The responses of the cells to the same instruction stimuli repeated twice in a row, and under the condition that the animal did not behaviorally use the first instruction in between, were tested. More than one-third of the tested cells (n = 14) did not respond, or responded very weakly, to the second instruction.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 7500162 TI - Do neurons from rat neostriatum express both a TTX-sensitive and a TTX insensitive slow Na+ current? AB - 1. The properties of a tetrodotoxin (TTX)-sensitive, persistent Na+ current and a purported TTX-insensitive slow Na+ current were studied in acutely isolated neurons from rat neostriatum with the use of the whole cell configuration of the patch-clamp technique. 2. A TTX-sensitive, persistent Na+ current (INaP) was activated positive to -60 mV and reached a peak amplitude of -40 to -120 pA at about -40 mV. As indicated by slow depolarizing voltage ramps, activation of INaP did not require preceding activation of the fast, rapidly inactivating Na+ current. 3. The current-voltage (I-V) relationship of INaP displayed an unexpected inflection after passing through its peak value near -40 mV. Between 40 and -10 mV, INaP declined more rapidly with depolarization than it did at more depolarized potentials. The corresponding conductance (GNaP) peaked at -40 mV and declined to a smaller limiting value at potentials positive to about -10 mV. 4. This behavior is not consistent with the notion that INaP arises solely from a bell-shaped window conductance that results from the overlapping steady-state activation and inactivation curves of the fast Na+ current in a narrow voltage range, nor with the notion that INaP is generated by a single uniform conductance independent of the fast Na+ current. 5. In addition to INaP, a second slow inward current (IS) was evoked when small monovalent cations were omitted from the internal solution. INaP and IS were present both in cells resembling medium spiny neurons and in cells resembling aspiny interneurons. 6. IS was insensitive to TTX (1.2 microM) and the Ca2+ channel blocker, cadmium.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7500164 TI - Adenosine facilitates in vivo neurotransmission in the superior colliculus of the rat. AB - 1. Electrical responses to volleys in afferent fibers in the optic tract were recorded in the superficial gray layer of anesthetized rat superior colliculus. A prominent negative wave with 4- to 6-ms peak latency in the upper part of the superficial gray layer and a sharp negative wave with 1.5- to 2-ms peak latency in the lower part of the superficial gray layer were elicited, corresponding to the C2 (upper part of the superficial gray layer) and the C1 (lower part of the superficial gray layer) postsynaptic potentials reported by Sefton. 2. These C1 and C2 waves were depressed by kynurenic acid or quinoxaline dione (DNQX) applied just beside the recording electrode, suggesting that neurotransmission in these pathways is mediated by glutamate. 3. Adenosine (10 microM) injected in the superficial gray layer enhanced both C1 and C2 potentials up to 170 and 140%, respectively. 4. Administration of a potent inhibitor of adenosine deaminase, erythro-9-(2-hydroxy-3-nonyl) adenine hydrochloride (EHNA; 5 mg/kg sc) increased the amplitudes of both C1 and C2 potentials to 125 and 130% of the initial levels, respectively. 5. The extracellular application of adenosine uptake inhibitors, dipyridamole (100 microM) and nitrobenzylthioinosine (NBI; 10 microM) also enhanced postsynaptic potentials. 6. Prior application of L-homocysteine thiolactone (10 microM), a compound that facilitates the incorporation of adenosine into S-adenosylhomocystein and reduces the extracellular concentration of adenosine, attenuated the excitatory action of exogenously applied adenosine. 7. Excitatory effects were also observed upon application of a selective adenosine A1 receptor agonist, N6-cyclohexyladenosine (CHA) or a selective A2 receptor agonist, 2-[4-(2-carboxylethyl)- phenethylamino]-5'N-ethylcarboxamide adenosine hydrochloride (CGS21680). Selective A1 and A2 receptor antagonists, 8 cyclopentyl-1,3-dimethylxanthine (CPT) and 3,7-dimethyl-1-propargylxanthine (DMPX), respectively, failed to suppress the excitatory action by adenosine. However, combined application of these two agents blocked the facilitatory action by adenosine on the excitatory synapses. 8. The application of adenosine (10 microM) to the superficial gray layer via a microdialysis probe increased the glutamate release by approximately 230% of the basal level. Similarly, the administration of EHNA (5 mg/kg sc) enhanced the extracellular glutamate level up to approximately 170%. However, prior application of L-homocysteine thiolactone (10 microM) failed to potentiate the glutamate release by adenosine. 9. This is the first in vivo study to demonstrate an excitatory action of adenosine on synaptic transmission.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7500163 TI - Calcium influx but not pH or ATP level mediates glutamate-induced changes in intracellular magnesium in cortical neurons. AB - 1. We have recently shown that glutamate increases [Mg2+]i in cultured rat cortical neurons. However, the mechanism of this increase in [Mg2+]i is not well understood. We used fluorescence microscopic methods to measure [Mg2+]i, [Ca2+]i, and pHi in single neurons. Intracellular ATP analysis was performed by high performance liquid chromatography (HPLC). 2. A 25-mM NH4Cl pulse followed by Na(+)-free wash rapidly acidified the cytosol. In 2',7'-bis(2-carboxyethyl)-5(6) carboxyfluorescein (BCECF)-loaded neurons, the pHi was reduced by 2.46 units, and in magfura-2-loaded neurons the [Mg2+]i was increased by 0.62 mM. Five-minute treatment with 100 microM glutamate, on the other hand, reduced the cytosolic pH by 0.73 units and increased the [Mg2+]i by 7.24 mM in rat cortical neurons. These results indicate that change in pHi does not play a significant role in the glutamate-induced [Mg2+]i elevation. 3. The metabolic inhibition (5 mM KCN and 1 mM iodoacetate) for 30 min significantly reduced the intracellular ATP levels. However, 5-min treatment with 100 microM glutamate did not significantly deplete intracellular ATP in cultured cortical neurons. When tested under similar conditions in magfura-2-loaded neurons, glutamate increased [Mg2+]i to a significantly larger extent than metabolic inhibition. This suggests that ATP depletion and subsequent release of Mg2+ from Mg(2+)-ATP complex is not the primary source of [Mg2+]i elevation observed during glutamate stimulation. 4. To further study the role of glutamate-induced Ca2+ influx in subsequent [Mg2+]i elevation, extracellular Ca2+ was elevated from 1.4 to 3.0 mM during glutamate application in magfura-2-loaded neurons.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7500165 TI - Comparison of noise and tone azimuth tuning of neurons in cat primary auditory cortex and medical geniculate body. AB - 1. A comparison of the azimuth tuning of single neurons to broadband noise and to best frequency (BF) tone bursts was made in primary auditory cortex (AI: n = 173) and the medial geniculate body (MGB: n = 52) of barbiturate-anesthetized cats. Observations were largely restricted to cells located within the tonotopically organized divisions of the MGB (i.e., the ventral nucleus and the lateral division of the posterior nuclear group) and the middle layers of AI. All cells studied had BFs > or = 4 kHz. 2. The responses of each cell to sounds presented from seven frontal azimuthal locations (-90 to +90 degrees in 30 degrees steps; 0 degree elevation) and at five sound pressure levels (SPLs: 0-80 dB or 5-85 dB in 20-dB steps) provided an azimuth-level data set. Responses were averaged over SPL to obtain an azimuth function, and a number of features of this function were used to describe azimuth tuning to noise and to tone stimulation. Azimuth function modulation was used to assess azimuth sensitivity, and cells were categorized as sensitive or insensitive depending on whether modulation was > or = 75% or < 75% of maximum, respectively. The majority (88%) of cells in the sample were azimuth sensitive to noise stimulation, and statistical analyses were restricted to these cells, which are presumably best suited to encode sound source azimuth. Azimuth selectivity was assessed by a preferred azimuth range (PAR) over which azimuth function values exceeded 75% (PAR75) or 50% of maximum response. Cells were categorized according to the location and extent of their noise PARs. Unbounded cells had laterally located PARs that extended to the lateral pole (+/- 90 degrees); bounded cells had PARs that were contained entirely within the frontal hemifield, and a subset of these had PARs centered on the midline (+/- 15 degrees). A final group of cells exhibited multipeaked azimuth functions to noise stimulation. 3. Azimuth functions to noise were generally more selective and/or more sensitive than those to tones. Statistical analyses showed that these differences were significant for cells in each azimuth function category, and for the thalamic and cortical samples. With the exception of multipeaked cells, responsiveness to noise was significantly lower than that to tones in all categories, and for the thalamic and cortical samples.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 7500166 TI - Fiber types contributing to dorsal root reflexes induced by joint inflammation in cats and monkeys. AB - 1. Injection of kaolin and carrageenan into the knee joint of cats or monkeys resulted in an acute inflammation. Four hours after injection of the knee joint, efferent activity could be evoked in articular afferent fibers and in dorsal root filaments. We interpret this efferent activity to be dorsal root reflexes (DRRs). Under our experimental conditions, the DRRs were generally synchronized compound action potentials, although in some cases single-unit activity was also observed. 2. DRRs were not produced in animals with uninflamed knee joints and normal body temperatures. 3. Recordings from two different sites on cut dorsal root filaments ipsilateral to the inflamed knee joint allowed the determination of the conduction velocities of groups of afferent fibers carrying DRRs. The DRRs occurred in A beta-, A delta-, and C fibers. However, in these experiments the peripheral destination of the afferent fibers was unknown. 4. To prove that DRRs occurred in joint afferents, recordings were made from two different sites on the proximal stump of the medial articular nerve that innervated the inflamed knee. The DRRs were again found in all fiber types, i.e., group II, III, and IV (A beta, A delta, and C) articular afferent fibers. 5. Compound DRRs were recorded from the central end of a cut dorsal root filament after electrical stimulation at C fiber intensity of a dorsal root adjacent to the filament. This DRR activity was eliminated by extensive dorsal rhizotomies of the L2-S1 roots.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7500167 TI - Cholecystokinin depolarizes rat thalamic reticular neurons by suppressing a K+ conductance. AB - 1. The thalamic reticular nucleus (nRt) is innervated by cholecystokinin (CCK) containing neurons and contains CCK binding sites. We used tight-seal, whole cell recording techniques with in vitro rat thalamic slices to investigate the action of CCK on neurons in nRt and ventrobasal thalamus (VB). 2. Brief applications of the CCK agonist cholecystokinin octapeptide (26-33) sulfated (CCK8S) evoked prolonged spike discharges in nRt neurons but had no direct effects on VB neuron activity. This selective excitatory action of CCK8S in nRt resulted from a long lasting membrane depolarization (2-10 min) associated with an increased input resistance. Voltage-clamp recordings revealed that CCK8S reduced membrane conductance by 0.6-3.8 nS, which amounted to 5-54% of the resting conductance of these neurons. 3. The conductance blocked by CCK8S was linear over the range of 50 to -100 mV and reversed near the potassium equilibrium potential. Modifications of extracellular K+ concentration altered the reversal potential of the conductance as predicted by the Nernst equation. The K+ channel blocker Cs+, applied either intracellularly or combined intra- and extracellularly, blocked the response to CCK8S. 4. The CCK8S-induced depolarization persisted after suppression of synaptic transmission by either tetrodotoxin or a low-Ca2+, high Mg2+ extracellular solution, indicating that the depolarization was primarily due to activation of postsynaptic CCK receptors and not mediated through the release of other neurotransmitters. 5. The selective CCKA antagonists L364,718 and Cam 1481 attenuated the CCK8S-induced depolarization, whereas the CCKB antagonist L365,260 had little or no effect on the depolarization. 6. Our findings indicate that CCK8S, acting via CCKA-type receptors, reduces a K+ leak current, resulting in a long-lasting membrane depolarization that can presumably modify the firing mode of nRt neurons. Through this effect, CCK actions in nRt may strongly influence thalamocortical function. PMID- 7500168 TI - Recent advances in methods of assessing dietary amino acid requirements for adult humans. AB - The requirements for the indispensable amino acids have been determined by a number of different methods. Historically, descriptive or gross measures like growth and nitrogen balance have been used. However, technological advancements in recent years have resulted in the use of more precise and mechanistic metabolic approaches (i.e., plasma amino acid concentrations, amino acid oxidation, indicator amino acid oxidation) to examine requirement. Nevertheless, the current recommendations are still based on nitrogen balance studies. Requirement estimates based on other methodologies, such as plasma amino acid concentrations and direct amino acid oxidation, suggest that the requirement estimates derived from nitrogen balance experiments are too low. However, these higher estimates have also been criticized on conceptual and methodological grounds, resulting in considerable controversy in the area of indispensable amino acid requirements. A new technique, indicator amino acid oxidation, addresses many of the criticisms directed toward the alternative methods and the proposed higher requirement estimates. This paper reviews the current knowledge of amino acid requirements and makes recommendations in light of new information that has been provided from recent indicator amino acid oxidation research. It is concluded that the nitrogen balance-based estimates of amino acid requirement are too low. PMID- 7500169 TI - Characterization of a novel protein in chick intestine that exhibits calcium binding activity and regulation by dietary calcium, aluminum and vitamin D. AB - The molecular mechanisms stimulated by vitamin D and low calcium diets that promote intestinal calcium absorption are not fully understood. In the present experiments, groups of chicks were subjected to the following treatments known to alter the efficiency of Ca absorption: vitamin D deficiency, repletion of vitamin D-deficient chicks with 1,25-dihydroxycholecalciferol [1,25(OH)2D3], low Ca intakes, and aluminum toxicity. Duodenal mucosal scrapings were obtained and screened for changes in protein composition using SDS-PAGE and size exclusion chromatography. The relative amount of a previously unreported 400-kDa oligomer containing 22-kDa monomeric subunits was found to vary directly with theoretical changes in the efficiency of Ca absorption, i.e., amounts increased with low Ca intakes and repletion with 1,25(OH)2D3, but were decreased by dietary aluminum and vitamin D deficiency. The oligomer was shown to have Ca-binding activity using a 45Ca overlay technique. These properties suggest 1) that the protein is regulated, at least indirectly, by 1,25(OH)2D3; 2) that the protein may play a role in promoting Ca absorption, possibly by binding Ca; and 3) that dietary aluminum interferes with the regulation of this protein, possibly by interfering with the actions of vitamin D in the intestine. PMID- 7500170 TI - Defects of copper deficiency in rats are modified by dietary treatments that affect glycation. AB - We examined the hypothesis that nonenzymatic glycosylatin of proteins (glycation) contributes to the defects of copper deficiency. We studied copper-adequate and deficient rats while altering two factors known to affect glycation: type of dietary carbohydrate and amount of food intake. Copper deficiency caused cardiac enlargement and anemia, decreased erythrocyte osmotic fragility, enhanced heart lipid peroxidation, increased the percentage of glycated hemoglobin (Hb A1) and reduced staining of lens crystallins on SDS-PAGE gels (suggestive of glycation). Increasing dietary sucrose reduced organ copper concentration, exacerbated the rise in Hb A1 and worsened the anemia caused by copper deficiency. Food restriction ameliorated heart and erythrocyte defects, reduced the percentage of glycated hemoglobin and heart peroxidation and also improved heart and liver copper status in copper-deficient rats. These findings indicate that copper deficiency enhances glycation and that sucrose may exacerbate some defects of copper deficiency by enhancing glycation. Inhibition of defects of copper deficiency by food restriction suggests that glycation and/or peroxidation may contribute to those defects. PMID- 7500171 TI - Copper deficiency increases hepatic apolipoprotein A-I synthesis and secretion but does not alter hepatic total cellular apolipoprotein A-I mRNA abundance in rats. AB - This study was designed to determine whether an increase in hepatic apolipoprotein A-I (apo A-I) synthesis and mRNA abundance is responsible for the enlarged plasma apo A-I pool observed in copper-deficient rats. Weanling male Sprague-Dawley rats were divided into two dietary treatments: copper-adequate (102.2 mumol Cu/kg diet) and copper-deficient (9.0 mumol Cu/kg diet). Copper deficiency resulted in a significant increase (124%) in intravascular apo A-I pool size after 6 wk of treatment. Following intraportal injection of a flooding dose of [3H]phenylalanine, in vivo hepatic apo A-I synthesis and secretion were significantly greater in the copper-deficient animals as detected by [3H]phenylalanine incorporation into immunoprecipitable apo A-I isolated from liver homogenates and plasma using anti-rat apo A-I antibodies. Pulse-chase experiments using freshly isolated hepatocytes demonstrated that a significant increase (148%) in apo A-I secretion by hepatocytes derived from copper-deficient rats may have resulted from increased hepatic synthesis rather than altered intracellular degradation of apo A-I. Hepatic total cellular apo A-I mRNA abundance was not altered by copper deficiency when expressed per microgram of RNA. Thus, the enhanced hepatic apo A-I synthesis, observed in copper-deficient cells, may have resulted from alterations in post-transcriptional and translational processes. PMID- 7500172 TI - Diet differentially regulates glucokinase and L-type pyruvate kinase gene expression in rat liver. AB - The regulation of gene expression of glucokinase (GK) and L-type pyruvate kinase (L-PK) in rat liver was investigated and compared with the previously reported regulation of lipogenic enzymes. Experiments were conducted in which the time courses and responses to diet quantity of mRNA concentrations and enzyme activities after refeeding a carbohydrate/protein diet (CP) to food-deprived rats were measured. The effects of dietary nutrients on the gene expression were investigated in rats refed either the CP diet, a carbohydrate diet without protein (C), a protein diet without carbohydrate (P), or a carbohydrate/protein/corn oil diet (CPF). The effects of the CPF diet on the gene expression after insulin treatment to diabetic rats were also investigated. After refeeding the CP diet, GK mRNA concentration and enzyme activity reached maximum levels in 2 h and 16-24 h, respectively, whereas those of L-PK peaked in 16 h and 48 h, respectively, similar timecourse to lipogenic enzymes. Moreover, GK mRNA concentrations were maximal in rats fed 20% of the ad libitum diet intake, and L PK mRNA concentrations, like lipogenic enzyme mRNA, were maximal in rats fed approximately 50% of ad libitum intake. GK mRNA concentrations were significantly increased in parallel with an increase in plasma insulin and glucose concentrations. GK and L-PK mRNA and enzyme levels in rats fed the C diet were comparably induced to the levels in those fed the CP diet. L-PK mRNA induction by the CP diet was significantly reduced by dietary polyunsaturated fatty acids (CPF diet), whereas the GK mRNA induction was not significantly reduced.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7500173 TI - CYP-2E1 inhibitors partially ameliorate the changes in hepatic fatty acid composition induced in rats by chronic administration of ethanol and a high fat diet. AB - The objective of this study was to determine if ethanol-induced cytochrome P450 2E1 (CYP2E1) was responsible for the changes in hepatic fatty acids observed in rats fed ethanol intragastrically. We hypothesized that if CYP2E1 was responsible for these changes then CYP2E1 inhibitors fed with ethanol should prevent the ethanol-induced changes in fatty acids. We compared the fatty acid composition of the liver in rats fed ethanol alone with that in rats fed ethanol with the CYP2E1 inhibitors, diallyl sulfide and phenethyl isothiocyanate. In each experiment, rats pair-fed isocaloric glucose were included to determine the effect of the inhibitors alone on the hepatic fatty acid composition. The lobular distribution of succinic dehydrogenase was determined histochemically because the lobular distribution of CYP2E1 shifts to the periportal area in livers of rats fed CYP2E1 inhibitors. The CYP2E1 inhibitors ameliorated both the ethanol-induced changes in fatty acids and the shift in succinic dehydrogenase. Rats fed ethanol but no inhibitors had significantly greater hepatic total fatty acids and triglyceride fractions than when inhibitors were fed ethanol. Ethanol altered the fatty acid composition compared with rats fed ethanol with CYP2E1 inhibitors. The ratio of 20:4/18:2 was significantly lower and that of 18:1/18:0 was greater in alcohol fed rats compared with their pair-fed controls. The CYP2E1 inhibitors inhibited many of the above effects of alcohol. The data suggest that the changes in the fatty acid composition due to ethanol ingestion are the result of CYP2E1 dependent lipid peroxidation and fatty acid metabolism. PMID- 7500174 TI - Randomized clinical trial of nutritional supplementation shows little effect on functional status among free-living frail elderly. AB - Nutritional deficiencies, frequently observed among the frail elderly, have been associated with increased morbidity and dependency. A randomized clinical trial was conducted to assess the impact of nutritional supplementation on the functional status of frail elderly living at home. Subjects (n = 50) over age 60 (mean 78 y) requiring community services and at elevated risk of undernutrition (excessive weight loss or body mass index < 24 kg/m2) were randomized to a high energy nutrient dense supplement provided by a dietitian or a control group receiving visits only. Outcome measures, including changes in handgrip strength, general well-being score, perception of health and number of falls, were measured by an investigator who was unaware of treatment assignment. After 12 wk, weight gain was greater in the supplemented group (2.1 +/- 2.3 vs. 0.6 +/- 1.6 kg; P < 0.01). Despite adequate statistical power, no functional measures differed except that the number of falls was lower among treated subjects vs. controls (0% vs. 21%; P = 0.05). Frail elderly can gain weight through oral supplementation, indicating the reversibility of weight loss in this group, but changes in functional status indicators may require a longer period of supplementation. PMID- 7500175 TI - Exclusive breast-feeding duration is associated with attitudinal, socioeconomic and biocultural determinants in three Latin American countries. AB - International health organizations have recommended exclusive breast-feeding (EBF) (i.e., breast milk as the only source of food) as the optimal infant feeding method during the first 4-6 mo of life. Therefore, it is important to document the determinants of EBF in different populations. Low-income urban women from Brazil (n = 446, 2 maternity wards), Honduras (n = 1582, 3 maternity wards) and Mexico (n = 765, 3 maternity wards) were interviewed at birth and in their homes at 1 mo and 2-4 mo after delivery. Multivariate survival analyses (Cox model) indicated that planned duration of EBF (all 3 countries), having a female infant, and not being employed (Brazil and Honduras), lower socioeconomic status (Honduras and Mexico) and higher birth weight (control hospital in Brazil and Honduras) were positively associated (P < or = 0.10) with EBF. Women who delivered in the maternity wards that had more developed breast-feeding promotion programs were more successful with EBF. The association between maternal education and EBF was modified by the maternity ward in Mexico and Honduras. Being > or = 18 y and having a partner living (Brazil) or not (Mexico) living at home were positively associated with EBF. These findings can contribute toward the design of EBF promotion efforts in Latin America. PMID- 7500176 TI - Vitamin B-6 status during childhood: tracking from 2 months to 11 years of age. AB - To examine the development and tracking of long-term vitamin B-6 status from infancy to early adolescence, measurements of erythrocyte pyridoxal 5'-phosphate concentration (EPLP), the erythrocyte aspartate transaminase (EAST) stimulation test including measurements of basal activity (EASTo) and activation coefficient (alpha EAST), were made in a follow-up study of healthy children aged 2 (n = 139), 4 (n = 147), 6 (n = 157), 9 (n = 159) and 12 mo (n = 188) and 5 y (n = 148). The EAST stimulation test was repeated at 11 y (n = 153). Vitamin B-6 status, high during infancy, reached the adult level by 5 y of age. The 10th to 90th percentile ranges for EPLP values were 61-201 nmol/L at 4 mo, 49-101 nmol/L at 12 mo and 27-59 nmol/L at 5 y. The respective ranges for Easto were 16-24 microkat/L at 4 mo, 13-19 microkat/L at 12 mo, 9-14 microkat/L at 5 y and 25-39 microkat/L at 11 y of age. For alpha EAST values were 1.29-1.54 at 4 mo, 1.48 1.77 at 12 mo, 1.70-2.07 at 5 y and 2.00-2.57 at 11 y. Values for EPLP and the EAST stimulation test in the first year of life correlated with the values at 5 and 11 y. The individuals with values at the extreme ends of the distributions remained there from infancy to childhood up to 3.3 times more often than expected from random variation.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7500178 TI - Supplemental nutrition with ornithine alpha-ketoglutarate in rats with cancer associated cachexia: surgical treatment of the tumor improves efficacy of nutritional support. AB - We investigated the use of ornithine alpha-ketoglutarate in treatment of rats bearing Morris hepatoma 7777. Rats received diets containing either ornithine alpha-ketoglutarate, which has been used in other catabolic states (i.e. injury, sepsis), or an isonitrogenous, isocaloric diet containing glycine. Untreated tumors grew to a mass of 11 g/100 g body weight over the 3-wk period after implantation and induced progressive anorexia, negative nitrogen balance, and body and tissue wasting. Compared with glycine, ornithine alpha-ketoglutarate had no effect on tumor growth, but also did not alter the catabolic effects of the tumor on its host. We hypothesized that capture of amino acids by the tumor limited the efficacy of supplemental nutrition here and in published reports in which tumor burden comprised 4-30% of body weight. This is supported by our observation that a 3-wk of implantation the rate of protein deposition plus amino acid oxidation by the tumor was equivalent to approximately 70% of the host's daily protein intake. To parallel the clinical situation in which tumor burden is small at diagnosis and initiation of treatment, the same diets were tested in rats treated by excision of the tumor at a limited stage of the disease. Rats received 3 d preoperative nutrition with ornithine alpha-ketoglutarate or glycine, and continued on the same diets for 3 or 6 d postoperatively. Compared with glycine-fed rats, ornithine alpha-ketoglutarate-fed rats showed a more positive nitrogen balance, higher concentrations of glutamine and branched-chain amino acids in muscle, and accelerated protein deposition in small intestine (P < 0.05). Our results explain the lack of success of nutritional support in untreated cancer and underline the need for clinically relevant animal models for further studies. PMID- 7500179 TI - Alterations of methionine fluxes and incorporation in intestines of miniature pigs fed a diet high in caseinate are restricted by angiotensin-converting enzyme inhibitor. AB - Previous results from our laboratory showed that a methionine-rich caseinate based (metcas) diet induces hyperhomocysteinemia in miniature pigs. In the present study, the contribution of the ileal and jejunal methionine absorption to the dietary induced hyperhomocysteinemia was evaluated by measuring the mucosal to serosal fluxes and the enterocyte incorporation in intact intestinal epithelia mounted in Ussing chambers. For 4 mo, 20 miniature pigs were daily fed control or metcas diets, and an oral combination of an angiotensin-converting enzyme inhibitor (25 mg captopril, Cp) and diuretic (12.5 mg hydrochlorothiazide, HTZ) or placebo, ileal incorporation was higher in epithelia from miniature pigs metcas than in that from other groups. For a given transepithelial flux of methionine, i.e., a constant amount of methionine recovered in the serosal chamber, a greater enterocyte incorporation was detected. Cp-HTZ treatment corrected the diet-induced methionine trapping in intestinal epithelia but had little effect in control animals. In separate in vitro experiments, Cp added alone significantly activated methionine fluxes in epithelia from metcas-fed miniature pigs as it did in vivo, demonstrating that Cp rather than HTZ mainly contributed to the in vivo effects of the drug combination. Our results showed that the regulation of intestinal methionine absorption compensated the diet induced hyperhomocysteinemia and that Cp-HTZ treatment altered these adaptative changes without increasing methioninemia and homocysteinemia. PMID- 7500180 TI - Whole body metabolism is not restricted to D-sugars because energy metabolism of L-sugars fits a computational model in rats. AB - Can L-sugars contribute to whole body energy metabolism? Energy balance studies were undertaken in rats fed L-sugars at a rate of 10 g/100 g basal diet. Partial metabolizable energy values (MEVp) during the last 28 of 56 d while consuming the L-sugar diets showed that L-glucose contributed no energy (0.3 +/- 0.9 kJ/g, P > 0.5, mean +/- SEM), whereas L-fructose and L-glucose contributed 6.9 +/- 0.9 (P < 0.001) and 8.8 +/- 1.8 (P < 0.001) kJ/g, respectively. Over periods from 0 to 28 and 0 to 56 d of L-sugar treatment, measurements were made of energy intake, the average lean mass of animals, fat and lean mass deposition. Using these measurements and a computational model, estimates were made of each supplement's net metabolizable energy value for maintenance (NEVs). These estimates confirmed the supply of energy from L-fructose, with NEVs of 5.6 +/- 3.2 (P < 0.32) and 6.1 +/- 1.7 (P < 0.01) kJ/g over 28 and 56 d, respectively, and L-gulose with NEVs of 10.2 +/- 3.4 (P < 0.01) and 11.4 +/- 1.8 (P < 0.001) kJ/g over 28 and 56 d, respectively. A lack of energy supply from L-glucose was confirmed with NEVs of 2.6 +/- 3.2 (P > 0.5) and 0.0 +/- 1.6 (P > 0.5) kJ/g over 28 and 56 d, respectively. As reference substrates, sucrose and cellulose gave expected energy values in all determinations. Methods comparison analysis showed no differences between MEVp and NEVS. The assimilation of energy from L-fructose and L-gulose is probably via large bowl microorganisms. These observations show that L-gulose and L-fructose contribute to whole body energy metabolism while L-glucose does not. PMID- 7500181 TI - Formation of second messenger diradylglycerol in murine peritoneal macrophages is altered after in vivo (n-3) polyunsaturated fatty acid supplementation. AB - The consequences of macrophage membrane incorporation of (n-3) polyunsaturated fatty acids (PUFA) on diradylglycerol (DG) formation and phospholipase-mediated signal transduction in vivo remain largely unknown. Three-to four-week-old C57BL/6 mice were randomly assigned to diets in which the 10% lipid component of the purified diet was either corn oil ethyl ester (control) or (n-3) ethyl ester [(n-3) PUFA-enriched]. After 4 wk, thioglycollate-elicited peritoneal macrophages were harvested and assayed for (1) total DG mass, (2) DG molecular species fraction analyses by argentation TLC and (3) separation of phospholipid classes and analyses of their molecular species fractions. We found that basal DG mass and the increase in DG mass after stimulation with platelet-activating factor or phorbol myristate acetate were similar in cells from control and (n-3) PUFA enriched diet groups, whereas ionomycin-stimulated DG formation was less in the cells from the (n-3) PUFA-enriched diet group. Analyses of DG molecular species fractions showed that the proportions of species with five and six double bonds were markedly greater in the (n-3) PUFA-enriched diet group macrophages compared with those of the control diet group. The molecular species fractions of all phospholipid classes including phosphatidylinositol showed substantial incorporation of (n-3) PUFA. These results demonstrate that dietary enrichment with (n-3) PUFA induces marked changes in DG metabolism in murine peritoneal macrophages. These changes may contribute to the mechanisms of antiinflammatory, antithrombotic and antineoplastic actions associated with dietary (n-3) PUFA supplementation. PMID- 7500177 TI - Milk composition in women from five different regions of China: the great diversity of milk fatty acids. AB - Human milk samples were obtained from 146 lactating women living in five distinct geographic regions of the People's Republic of China. The regions were characterized as follows: pastoral, rural, urban 1, urban 2, and marine. Dietary information obtained on a subsample of women providing milk suggested distinct differences in the food availability in the five regions and the habitual diets of the lactating women studied. The milk concentration of docosahexaenoic acid (DHA) varied greatly (g/100 g total fatty acids): 0.44 +/- 0.29 (pastoral), 0.68 +/- 0.29 (rural), 0.88 +/- 0.34 (urban 1), 0.82 +/- 0.35 (urban 2), and 2.78 +/- 1.20 (marine region). Docosahexaenoic acid is critical for brain and retina development in the infant. Milk arachidonic acid (AA) ranged from 0.80 +/- 0.14 (rural) to 1.17 +/- 0.33 and 1.22 +/- 0.32 g/100 g fatty acids from the marine and pastoral regions. Arachidonic acid is associated with infant growth. The AA to DHA ratio (g/g) was highly variable also: 2.77 (pastoral), 1.18 (rural), 1.01 (urban 1), 1.23 (urban 2), and 0.42 (marine). The DHA concentration found in the milk of women from the marine region was twice as high as any reported previously, but was in a range similar to the amounts found in the milk of women fed fish oil. Seafood consumption by the women residing in the marine region is a likely contributor to the DHA concentration in their milk. These data illustrate the heterogeneity of the fatty acid composition of human milk and suggest a great influence of geography and the maternal diet on concentrations of DHA and AA in human milk. PMID- 7500183 TI - Pregnancy and lactation are associated with diminished concentrations of choline and its metabolites in rat liver. AB - Choline is an important nutrient that is actively transported from mother to fetus across the placenta and from mother to infant across the mammary gland. Thus, pregnancy and lactation are times when dietary requirements for choline may be increased. Pregnant rats eating AIN-76A diet (with and without choline) for 6 d (d 12-18 gestation) were compared with nonmated female and male rats eating the same diets. Similarly, lactating rats were compared with nonmated female rats, both groups eating these same diets for 25 d (gestation d 12-postpartum d 15). We measured choline and choline metabolites in livers on the last day of feeding. Nonmated female rats, eating the control diet, had higher hepatic choline metabolites concentrations than did male rats (choline, 98%; betaine, 96%; and phosphorylcholine, 55% higher), pregnant rats (phosphorylcholine, 47%; and betaine, 42% higher) or lactating rats (phosphorylcholine, 49%; phosphatidylcholine, 37%; and betaine, 273% higher). We found that nonmated females eating a choline deficient diet had only a modest diminution (33%) of the labile choline metabolite PCho in liver, compared with similar rats eating a control diet. When compared with similar rats fed a choline-adequate diet, pregnant rats fed a choline-deficient diet had significantly great diminution of hepatic phosphorylcholine (83% lower) than did nonmated females. Liver phosphorylcholine was only 12% lower than in controls in nonmated females fed the deficient diet for the same 25-d period. Lactating rats were the most sensitive to choline deficiency, with liver phosphorylcholine 88% lower than in similar rats fed control diet.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7500182 TI - Coconut oil and beef tallow, but not tricaprylin, can replace menhaden oil in the diet of red drum (Sciaenops ocellatus) without adversely affecting growth or fatty acid composition. AB - The ability of juvenile red drum (Sciaenops ocellatus) to utilize medium-chain triglycerides (MCT) and other saturated dietary lipids was investigated in two 6 wk feeding experiments. Diets contained solvent-extracted menhaden fish meal to which menhaden fish oil (control), coconut oil, corn oil, beef tallow or various levels of MCT as tricaprylin (30, 46, 65 and 80% of total lipid) were added. Diets were fed to triplicate groups of juvenile red drum in aquaria containing brackish (6%) water. In the first feeding experiment, red drum fed the control diet had the greatest weight gains and feed efficiencies. Weight gain, but not feed was slightly, of fish fed corn oil and fish fed coconut oil was slightly (P < 0.05) lower. In the second feeding experiment, fish fed coconut oil and those fed beef tallow had significantly higher weight gains and feed efficiencies than did fish fed the control diet. Fish fed the diets containing tricaprylin at all inclusion levels in both feeding experiments had significantly lower weight gains and feed efficiencies and higher levels of beta-hydroxybutyric acid in plasma. Fish fed diets with high levels of MCT also had lower (n-3) and greater (n-6) fatty acid levels in the neutral lipid fraction of muscle tissue compared with fish fed the control diet. Coconut oil and beef tallow consistently resulted in greater liver lipid deposition but had variable effects on other tissue indices. Saturated dietary lipids had variable effects on fatty acid composition of muscle polar and neutral lipid fractions.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7500184 TI - Ascorbate polyphosphate is a bioavailable vitamin C source in juvenile rainbow trout: tissue saturation and compartmentalization model. AB - We studied the bioavailability of ascorbic acid ester, ascorbate polyphosphate, to juvenile rainbow trout (Oncorhynchus mykiss). Fish were fed molar equivalents of 0, 20, 40, 80, 160, 320 and 1280 mg ascorbic acid/kg diet in the form of ascorbate polyphosphate. During the 18 wk of the experiment, when body weight increase averaged 3.5-fold, we did not observe any deficiency symptoms in any group. Liver and kidney ascorbate concentrations differed significantly among groups after wk 9. The ascorbic acid concentrations in liver were significantly different in fish fed for 9 wk an equivalent of 0, 40 and 160 mg ascorbic acid/kg as ascorbate polyphosphate, values were 22.7 +/- 3.4, 93.7 +/- 17.0 and 368.0 +/- 60.8 nmol ascorbic acid/g. The ascorbic acid concentrations in kidney were significantly different in fish fed for 18 wk an equivalent of 0,20 and 40 mg ascorbic acid/kg as ascorbic polyphosphate (23.9 +/- 4.0, 72.1 +/- 13.6 and 254.4 +/- 22.7 nmol ascorbic acid/g, respectively). After wk 18, fish from groups fed 0, 20, 320 and 1280 mg ascorbic polyphosphate/kg were intraperitoneally injected with 25 mg/ascorbic acid/kg body wt. We observed differences in the profiles of tissue ascorbate concentration during the 96 h following the injection between groups with high and low tissue ascorbate concentration, i.e., fish fed 320 and 0 ascorbic acid/kg, respectively. We conclude that ascorbic acid metabolism in rainbow trout after intraperitoneal injection followed the three-compartmental model, with the intraperitoneal cavity as the first compartment, blood as the second, and tissues as the third.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7500186 TI - Tissue alpha-tocopherol retention in male rats is compromised by feeding diets containing oxidized frying oil. AB - To investigate the effect of dietary oxidized frying oil (OFO) on tissue retention of alpha-tocopherol (alpha-T). Long-Evans male weanling rats were divided to four groups based on a 2 x 2 factorial design. Two groups were fed 15% OFO diets, and the remaining two groups were fed control diets in which OFO was replaced by vitamin-E-stripped fresh soybean oil. Vitamin E as all-rac-alpha tocopheryl acetate was added at the concentration of either 50 (normal E) or 500 (high E) mg/kg diet. The OFO sample was prepared by deepfrying sticks in fresh soybean oil at 205 +/- 5 degrees C for four 6-h periods. After 6 wks of feeding, alpha-T concentrations in most tissues were significantly lower in rats fed OFO diets (P < 0.05) than in the control groups. For rats fed the OFO diet with the normal vitamin E concentration, the alpha-T concentration is epididymal fat pad, plasma, liver, kidney, muscle, brain and lung were 29-64% those of the corresponding control group (P < 0.05). The interaction between the two dietary factors on tissue alpha-T was significant in liver, spleen, and adrenal gland. In these three tissues, the differences between the normal and high dietary vitamin E groups were less in rats fed the OFO diets than in rats fed the control diets. The tissue alpha-T concentrations of the high vitamin E OFO group were comparable with or higher (P < 0.05) than those of the normal vitamin E control group, indicating that the negative effect of OFO on tissue alpha-T concentration can be alleviated by dietary supplementation of vitamin E. Compared with the controls, rats OFO diets had significantly higher tissue thiobarbituric acid reactive substances (P < 0.05). Because the amount of alpha-T directly added into the test oil samples was not significantly decreased through an incubation (at 37 degrees C) period of up to 10 d, the inefficient absorption and/or enhanced catabolism or turnover of vitamin E may be involved in the inferior tissue alpha-T retention of OFO fed rats. PMID- 7500185 TI - Dietary polyunsaturated fatty acids and aging modulate glutathione-related antioxidants in rat liver. AB - The effects of dietary polyunsaturated fatty acids (PUFA) on the age-dependent changes in liver glutathione antioxidant system were investigated in male Wistar rats fed diets supplying either balanced amounts of linoleic acid and alpha linolenic acids (control) or deficient in alpha-linolenic acid [n-3) deficient]. The animals were studied at the age of 6 or 24 mo. Glutathione antioxidative metabolism was markedly affected by aging. Cytosolic concentration of reduced glutathione (GSH) was lower (P < 0.01), whereas that of oxidized glutathione (GSSG) as well as the GSSG:GSH ratio were greater (P<0.001) in the 24-mo-old as compared with the 6-mo-old rats, regardless of the diet. Glutathione peroxidase activities were lower (P < 0.001) in 24-mo-old rats, though more markedly in those fed the control diet. The lipid composition of rat liver microsomal membranes was strongly modified by both diet and aging. The age-related changes mainly involved the polyunsaturated fatty acids. These results suggest that the nature of dietary PUFA and not only their degree of unsaturation affects the cellular glutathione-dependent antioxidant system and thus may modify the age related changes in metabolic reactions. PMID- 7500187 TI - Simultaneous dietary supplementation of sodium cholate and beta-carotene markedly enhances accumulation of beta-carotene in mice. AB - This study evaluated whether simultaneous supplementation of sodium cholate and beta-carotene to a diet enhanced the accumulation of beta-carotene in mice. For 2 wk, male ICR mice were fed either a basal diet or a diet containing Dunaliella bardawil beta-carotene 50 mg/100g that was or was not supplemented with sodium cholate (0.25 g/100 g). The concentrations of beta-carotene in liver and plasma were approximately 5 and 10 times higher, respectively. In the mice fed the beta carotene diet with sodium cholate than in those fed the beta-carotene diet without sodium cholate. Beta-carotene was not detectable in the liver or plasma of mice fed either basal diet. The concentrations of vitamin E in the plasma and liver of mice fed either beta-carotene diet or the basal diet with sodium cholate were significantly lower than in those fed the basal diet. In a second study, mice were fed a diet containing 50 mg/100 g synthetic beta-carotene supplemented with various concentrations of sodium cholate (0, 0.05, 0.1, 0.25, 0.5 g/100 g) for 2 wk. The concentrations of beta-carotene and vitamin E in plasma, liver and bone marrow cells were higher in mice fed the beta-carotene diet supplemented with 0.05 g/100 g of sodium cholate than in those fed the unsupplemented diet. These findings show that simultaneous supplementation of sodium cholate and beta carotene to a diet markedly enhances the accumulation of beta-carotene. This dietary protocol may be useful to introduce a high amount of beta-carotene in the tissue of mice in a short period of time. PMID- 7500188 TI - Nursing education. PMID- 7500190 TI - The perinatal nurse practitioner: an innovative model of advanced practice. AB - When a major urban hospital received notice it would no longer have resident physicians to serve its perinatal population, nurses and physicians joined forces with a private university to develop a curriculum for advanced level caregivers. This willingness to rise to a challenge helped to create the Perinatal Nurse Practitioner Program and has paved the way for other needed inpatient nurse practitioner roles. PMID- 7500189 TI - Perinatal nurse practitioners and health care reform. AB - Health care delivery for pregnant women at high risk is changing in response to the nation's need to contain costs and improve care. Perinatal nurse practitioners can provide specialized care to women at high risk in a wide variety of settings. They provide quality, cost-effective care and can improve access to prenatal care for families at risk for untoward pregnancy outcomes. PMID- 7500191 TI - Women's reports of breast implant problems and silicone-related illness. AB - OBJECTIVE: The safety of silicone breast implants recently has been questioned. Increasing numbers of women have reported a variety of health problems that they attribute to their implants. The purpose of this descriptive study was to explicate the phenomenon of silicone-related illness as reported by women who have had breast implants. DESIGN: A qualitative design was grounded in the theoretical framework proposed by McBride and McBride (1981), the core of which is the first-person, lived experiences of women as interpreted and explained by the women themselves. PARTICIPANTS: Participants were recruited through Command Trust Network, an international support group for women with breast implant problems. The sample consisted of 55 women from 19 states and Canada who experienced health problems they attributed to their implants. Eighteen percent labeled themselves as disabled. RESULTS: Themes emerging from responses to 10 questions are discussed in terms of circumstances leading to initial implantation, understanding risks and benefits, health problems and symptoms, physician response, choices made about implant removal, and psychosocial and emotional consequences. CONCLUSIONS: Findings included dissatisfaction with implants, similarity of health problems, and recurrent surgical and nonsurgical procedures. Dominant themes included lack of informed consent, physician denial of health problems, and the decision to remove implants. Further research should be directed at recipients of implants who report silicone-related illness to determine the number of women affected and the nature of their problems. PMID- 7500192 TI - Creating paths: living with a very-low-birth-weight infant. AB - OBJECTIVE: To describe parents' method of adaptation to the problems of caring for a very-low-birth-weight (VLBW) infant at home. DESIGN: Exploratory, involving three interview sessions, 1 month, 3 months, and 5 months after hospital discharge of a VLBW infant. SETTING: A newborn intensive-care unit in a tertiary care center. PARTICIPANTS: Fourteen parents of VLBW (< 1,500 g) infants after hospital discharge. RESULTS: A basic social process, Creating Paths, was identified. It is the continuous process experienced by parents living with a VLBW infant the first 5 months after hospital discharge. The process consists of three stages: gathering, emerging, and affirming. CONCLUSIONS: This grounded theory systematically tracks and describes the adaptation process of parents with VLBW infants during the first 5 months of living at home. The theory may be used to anticipate concerns and provide guidance and support to parents and their infants. PMID- 7500193 TI - Physiology of the neonatal immune system. AB - Essential to a neonate's survival is an ability to respond effectively to hostile environmental forces. The developing neonate's immune system has functional, albeit limited, defensive, homeostatic and surveillance capabilities. The development of the cellular components of the neonatal immune system and their physiologic interrelationships and limitations are discussed. PMID- 7500194 TI - Bacterial and fungal infections in the neonate. AB - Despite tremendous advances in technology, sepsis remains a major threat in neonatal intensive care. The incidence of neonatal infection ranges from 7.3 to 16 per 1,000 live births. Nurses play an important role in the recognition and treatment of the septic infant. An understanding of the classification, epidemiology, and signs and symptoms of bacterial and fungal sepsis will better position the nurse to identify the infant at risk for sepsis and to initiate supportive care. PMID- 7500195 TI - Pharmacologic treatment of neonatal sepsis: antimicrobial agents and immunotherapy. AB - Antimicrobial agents and immunotherapies in the management of neonatal sepsis are discussed. The rationale for the selection of the most commonly used antibacterial drugs, their mechanisms of actions, and indications for use are described. Immunotherapies, both those in clinical use and those under investigation, are discussed. Antibacterial drugs remain the standard of care, but immunotherapy offers the potential for improving outcomes for neonates, especially those who are premature. PMID- 7500196 TI - Viral infections of the newborn. AB - Viral infections of the newborn result in significant morbidity and mortality each year. The fetus and newborn are particularly vulnerable to viral infection. The range of expression may vary from no clinical disease to devastating illness and infection occurring before, during, or after birth. Nursing management is determined by the specific viral infection, the severity of the illness, and the unique conditions of the newborn and his/her family. Promising new therapies are on the horizon that may lessen the severity of viral disease. Until such time, the major thrusts of management of neonatal viral disease are prevention of infection and supportive care for the acutely ill newborn. PMID- 7500198 TI - The immunologic significance of breast milk. AB - The importance of breast milk in protecting the newborn from infection is recognized worldwide. Infant morbidity and mortality have been directly affected by a decline in breastfeeding. Health care providers are working toward meeting the national goal of increased initiation and duration of breastfeeding. This article focuses on the protective factors transferred to the infant through breast milk. A discussion of maximizing the immunologic benefits of breast milk for the high-risk infant is presented. PMID- 7500197 TI - Immunization and the prevention of childhood diseases. AB - Nursing has an important role in improving the incidence of childhood immunization. An understanding of the immunologic deficiencies of children, especially those younger than 5 years, demonstrates the need for this population wide protection. Knowledge of the various products, their efficacy, schedule, side effects, and contraindications will assist the neonatal and pediatric nurse in coordinating immunizations and teaching parents about this important health priority for their children. Research is focusing on the development of combination vaccines and products for infectious diseases that currently have difficult or nonexistent treatment. PMID- 7500199 TI - Dependence of visual latency on wavelength: predictions of a neural counting model. AB - Simple reaction time (RT) was determined as a function of wavelength for equally visible, near-threshold stimuli. The stimuli were 5-deg spectral onsets of 1,000 ms duration presented on a 100-Td spatially coincident white background. All three subjects manifested the same result: RTs were fastest in the region of 590 nm. These data were analyzed in the context of a counting model of visual latency. This model predicts that for equally visible stimuli a transient detector will result in shorter visual latencies than will a more sustained detector. On the basis of this analysis, it is concluded that although most long duration, near-threshold, spectral step onsets are detected by the sustained parvocellular pathway, an exception occurs in the region of 590 nm: these stimuli are detected by a relatively transient pathway, presumably the magnocellular pathway. PMID- 7500200 TI - Differences between stereopsis with isoluminant and isochromatic stimuli. AB - Contrast thresholds for stereoscopic depth identification (crossed or uncrossed) were measured as a function of disparity by use of isoluminant (red-green) and isochromatic (yellow-black) 0.5 cycles/deg Gabor patches. For the purposes of comparison, stimulus contrasts were scaled by their respective detection thresholds. The Gabor patches could be either vertically or horizontally oriented. It was found that the disparity dependence of the depth-identification contrast thresholds was similar for both chromatic and luminance patterns if the stimuli were vertically oriented, with the overall level of performance worse for the chromatic patterns by a factor of approximately 2 (6 dB). With horizontal patterns this difference was much larger, by a factor of approximately 7 (17 dB). These results suggest first that stereopsis in the absence of luminance cues is supported by a less-contrast-sensitive linear mechanism than that which supports stereopsis in the presence of luminance cues and second that the corresponding nonlinear chromatic stereo mechanism is either nonexistent or very weak. The implications of these data for previous studies of stereopsis at isoluminance is discussed. PMID- 7500201 TI - Representation of videokeratoscopic height data with Zernike polynomials. AB - Videokeratoscopic data are generally displayed as a color-coded map of corneal refractive power, corneal curvature, or surface height. Although the merits of the refractive power and curvature methods have been extensively debated, the display of corneal surface height demands further investigation. A significant drawback to viewing corneal surface height is that the spherical and cylindrical components of the cornea obscure small variations in the surface. To overcome this drawback, a methodology for decomposing corneal height data into a unique set of Zernike polynomials is presented. Repeatedly removing the low-order Zernike terms reveals the hidden height variations. Examples of the decomposition and-display technique are shown for cases of astigmatism, keratoconus, and radial keratotomy. PMID- 7500202 TI - Discrimination of binocular color mixtures in dichromacy: evaluation of the Maxwell-Cornsweet conjecture. AB - We tested the Maxwell-Cornsweet conjecture that differential spectral filtering of the two eyes can increase the dimensionality of a dichromat's color vision. Sex-linked dichromats wore filters that differentially passed long- and middle wavelength regions of the spectrum to each eye. Monocularly, temporal modulation thresholds (1.5 Hz) for color mixtures from the Rayleigh region of the spectrum were accounted for by a single, univariant mechanism. Binocularly, univariance was rejected because, as in monocular viewing by trichromats, in no color direction could silent substitution of the color mixtures be obtained. Despite the filter-aided increase in dimension, estimated wavelength discrimination was quite poor in this spectral region, suggesting a limit to the effectiveness of this technique. PMID- 7500203 TI - Color matches in diseased eyes with good acuity: detection of deficits in cone optical density and in chromatic discrimination. AB - Reduced foveal cone optical density in diseased eyes with normal acuity can affect color matches. Using field diameters of 1 degree, 2 degrees, 4 degrees, and 8 degrees, we measured mean color-match midpoints and match widths in patients who had good acuity and who exhibited three categories of eye disease: hereditary macular degeneration (n = 12), retinitis pigmentosa (n = 19), and glaucoma (n = 18). Results were compared with those for normal observers of comparable ages. Mean color-match midpoints were abnormal only for the population with hereditary macular degeneration, indicating a reduction in cone optical density in the central 4 degrees. Mean color-match widths were enlarged for both hereditary macular degeneration and retinitis pigmentosa, a result consistent with a reduction in the number of foveal cones. PMID- 7500204 TI - Eigenvector interpretation of the Farnsworth-Munsell 100-hue test. AB - We measured the reflectance spectra for the 85 color caps of the Farnsworth Munsell 100-hue test. Eigenvectors and eigenvalues of a correlation matrix of cone responses were computed, with the cone responses being determined from the 85 test caps, arranged in order (according to color) by means of a linear model. It is shown that the Farnsworth-Munsell 100-hue test can be simulated by use of eigenvectors of the cone responses. The eigenvectors can be interpreted as nonopponent signal and opponent color signals. The normal observer can determine the color of a cap by using two opponent color signals. For color-blind persons (dichromats) one or the other opponent signal is defective, and errors can occur during the test. The simulation results also suggest that eigenvectors can be used to predict results of arrangement tests similar to the Farnsworth-Munsell 100-hue test. PMID- 7500207 TI - Rod transduction parameters from the a wave of local receptor populations. AB - The analysis of electroretinogram a waves from locally stimulated populations of rods is complicated by the presence of scattered light within the eye. Scattered light and cone contributions can be assessed after brief flashes of light designed to saturate only rods in the locally stimulated area. Subtracting the scattered-light and the cone responses from the local electroretinogram gives a pure rod a wave that can be fitted with models of photoreceptor activity. We demonstrate the feasibility of this technique by recording local rod a waves from a group of five normal subjects and by fitting the a waves with the rod model to derive transduction parameters. The local rod a waves are compared with expected responses derived from simulations in which the response of the entire retina to heterogeneous illumination is mimicked. PMID- 7500206 TI - Disability glare: effects of temporal characteristics of the glare source and of the visual-field location of the test stimulus. AB - One of the main early complaints of cataract patients, even when these patients exhibit only mild glare problems as measured by standard tests, is that glare impairs their night driving. To provide a better measure of the patients' impairment, glare tests should include measurements of the glare effect in conditions more similar to those found in night driving. During night driving the ambient light is very low, and oncoming headlights present a transient temporal pattern. Furthermore, the objects of interest often appear initially in the peripheral visual field. Thus three important characteristics of glare in night driving are that the ambient illuminance is in the scotopic-mesopic range, the detection stimulus is in the periphery, and the glare source is transient. Most of the current glare testers measure glare only at photopic levels, and all the glare tests that we know of use only steady sources of glare with foveal discriminations. All these conditions are dealt with. The transient glare source raised thresholds by 0.5-0.75 log unit more than the steady glare source, and the transient glare effect was more pronounced and more long lasting in the periphery. Standard glare testers seriously underestimate disability glare effects in everyday life. PMID- 7500205 TI - Effects of luminance and spatial noise on interferometric contrast sensitivity. AB - Optical properties of the eye contribute to the reduced visibility of spatial patterns at low luminance. To study the limits of spatial vision when optical factors are minimized, we measured contrast-sensitivity functions (CSF's) for 543.5-nm laser interference fringes imaged directly on the retina. Measurements were made in the fovea at four luminance levels, ranging from 0.3 to 300 photopic trolands (Td). At each luminance the fraction of coherent light in the stimulus pattern was varied to assess the masking effects of laser speckle, which is visible as spatial noise in fields of coherent light. Compared with published CSF's obtained under natural viewing conditions, interferometric CSF's were similar in height but broader, with the range of visibility being extended to higher spatial frequencies. The masking effects of speckle were greatest at the highest luminance and were negligible at the lowest luminance. For low coherent fractions, contrast sensitivity improved over the entire luminance range at a rate consistent with a square-root law; with purely coherent light, sensitivity tended to level off at approximately 30 Td because of speckle masking. The results indicate that the optical quality of the eye reduces the spatial bandwidth of vision even at luminances near the foveal threshold. The change in interference fringe visibility with luminance is consistent with noise-limited behavior, and the masking effects of speckle noise diminish as luminance decreases. PMID- 7500208 TI - Aberrations associated with rigid contact lenses. AB - A rigid contact lens on an eye can produce levels of spherical aberration very different from those produced by a spectacle lens in front of the eye. These levels are considerably affected by contact lens surface asphericity. Change in longitudinal spherical aberration associated with aspherizing a contact lens surface is well predicted by a simple equation for change in sagittal power of the surface. Displacing an aspheric contact lens on the eye can produce considerable defocus, which is well predicted by simple equations for change in sagittal and tangential surface powers. The best refractive correction with contact lenses can be determined only by overrefraction with a patient wearing a contact lens of power and characteristics similar to that which will be prescribed. An aspheric contact lens that moves to a considerable extent on the eye will cause more unstable vision than will a spherical lens that moves to the same extent. PMID- 7500211 TI - Performance on three-point vernier acuity targets as a function of age. AB - With certain (but not all) vernier acuity (VeA) stimulus displays observers exhibit no or slight performance decrements with aging. Because three-point VeA test displays are much more easily applied experimentally and clinically, we examined visual performance on this task as a function of age. Ages of groups tested ranged from 20 to 70+ years (some individuals were 90+ years old). Eyes were examined monocularly. We tested three- and two-point sharply defined VeA arrays and a three-point ground-glass-degraded display (simulating a 20/100 nuclear cataract). Gap or feature separations were varied. Results obtained with the three-point display showed little alteration with age. Image degradation resulted in reduction in performance when small gap separations were used between the test points, but only slight decrements for large gap separations were recorded. The finding that performance with certain VeA targets is largely unaffected by age is confirmed and is expanded. PMID- 7500210 TI - Scrolled and rapid serial visual presentation texts are read at similar rates by the visually impaired. AB - Visually impaired observers read dynamically displayed text faster than text displayed in a normal page view. The goal of this study was to compare reading rates from two dynamic-presentation methods that have been proposed to facilitate reading from computer-based displays. Prior research has shown that both normally sighted and low-vision observers read text displayed to the same location, one word at a time [known as rapid serial visual presentation (RSVP)], faster than a page of text. A similar comparison with text scrolled continuously across the screen also shows faster reading for low-vision patients, but the relative change from a standard page view is substantially less (15% faster for the scroll display versus 80% faster for RSVP). In this study we directly compared these techniques. For those with normal vision, reading from the RSVP display was 1.3 times faster than reading from the scroll display [t(9) = 3.32, P = 0.009]. Although the difference in reading rates for the visually impaired group did not reach statistical significance, as a group they read 13% slower from the RSVP than from the scroll display. PMID- 7500209 TI - Fiber-optic reading magnifiers for the visually impaired. AB - We describe fiber-optic stand magnifiers specifically designed for use as low vision reading aids. Application of this technology results in better optical and ergonomic properties. The fiber-optic magnifiers (tapers) provide bright, uniformly illuminated, distortion-free images. The reading material can be scanned without the user's having to bend directly over the magnifier. One can further increase the scanning field by slanting the taper to tilt the image toward the observer. Tilting the upper face of the taper by cutting the smaller lower face on a bias is shown to increase the scanning range substantially and to provide better control of illumination. The scanning range of such tilted tapers is approximately double that of the equivalent lens magnifiers. To increase the contrast transfer through the magnifiers, we have developed lower-resolution tapers with an increased core-to-cladding ratio. The increase in contrast transfer is reported for representative tapers. The lower-resolution design is also helpful in reducing the manufacturing cost of taper magnifiers. PMID- 7500212 TI - Spatial alignment across gaps: contributions of orientation and spatial scale. AB - To assess the contributions of orientation and spatial scale to the processing of relative-position information for broadband spatial targets, we measure misalignment thresholds for dots separated by as much as 6 deg, in the presence of one-dimensional spatial noise. For all the dot separations, thresholds for misalignment are raised most when the mask is oriented at approximately 20 deg to either side of true alignment. This bimodal orientation tuning function appears to be fundamental to the alignment judgment, including abutting vernier acuity for equally visible lines [Vision Res. 33, 1619 (1993)]. With increasing dot separation the spatial frequency at which peak masking occurs becomes progressively lower, a finding that suggests that the spatial mechanisms important for processing this information become larger. However, the rate of increase in size of these putative mechanisms is insufficient to account for the increase in relative-position thresholds for increasingly separated stimuli (i.e., Weber's law for alignment). In addition, oriented masks placed between two target lines lead to threshold elevation, revealing that the collection of positional information between target features may be important for optimal processing of misalignment thresholds. The findings of this study suggest that, although shifts in spatial scale of the underlying low-level oriented mechanisms may contribute to increased misalignment thresholds with increasing separation, additional factors, such as positional uncertainty associated with eccentricity per se, are limiting. PMID- 7500213 TI - Foveal adaptation abnormalities in early glaucoma. AB - Foveal sensitivities were measured after onset of adapting background fields for each of the following four groups of subjects aged 40-70 years: (1) low-tension glaucoma subjects with minimal field loss in the test eye, (2) primary open-angle glaucoma subjects with minimal field loss in the test eye, (3) normal control subjects, and (4) subjects originally enrolled as control subjects but subsequently found, on the basis of masked clinical evaluation, to be suspect for glaucoma despite ostensibly normal intraocular pressures. We found that the desensitization of a short-wavelength-sensitive-cone-mediated response after onset of a 580-nm background field was diminished from that of normal observers for low-tension glaucoma subjects but not for primary open-angle glaucoma subjects. The desensitization was also diminished for a glaucoma-suspect subjects aged 60-70 years. In contrast, the flicker sensitivity instabilities that persisted after onset of a long-wavelength background field for the majority of subjects with primary open-angle glaucoma [J. Glaucoma Suppl. 3, S19 (1994)] occurred only infrequently among the other subject groups. These results imply that glaucoma often involves the fovea, probably by affecting retinal subtractive adaptation processes, although with different consequences for different types of glaucoma. The results also suggest that undiagnosed low-tension glaucoma may not be rare in the general aging population. PMID- 7500215 TI - Incoherent spatial impulse response in variable-cross-section photoreceptors and frequency-domain analysis. AB - The spatial impulse response of a single variable-cross-section photoreceptor has been characterized assuming the incoming radiation to be an initial field propagating under confinement conditions inside the receptor. By application of Fourier analysis the total transfer function is determined. Both the outer and the inner segments are characterized as low-pass filters. The dependence of the transfer function on the modal parameter is analyzed. PMID- 7500214 TI - Direct measurement of human-cone-photoreceptor alignment. AB - We have developed an imaging reflectometer to measure cone-photoreceptor alignment. One makes measurements by bleaching the cone photopigment and imaging the distribution of light returning from the retina, which is illuminated from a small source imaged in the plane of the eye's pupil. If the source is near the optimal entry pupil position as determined psychophysically, the distribution of light returning from the retina is peaked, and the magnitude of the peak depends on the location of the source in the pupil of the eye. If the source is far from the optimal entry pupil position, then there is no measurable peak. The location of the peak varies across individuals and coincides with the reported location of best visibility of the measuring light and with previous psychophysical and reflectometric measurements of the Stiles-Crawford peak. The source of this directionality must arise either from the photoreceptors or from behind the photoreceptors because the peak is not present if measurements are made when the cone photopigments have high optical density. PMID- 7500217 TI - Double-pass measurements of the retinal-image quality with unequal entrance and exit pupil sizes and the reversibility of the eye's optical system. AB - We have used a modified double-pass apparatus with unequal entrance and exit pupil sizes to measure the optical transfer function in the human eye and have applied the technique to three different problems. First, we confirm that in the eye the double-pass spread function is the cross correlation of the input spread function with the output spread function [J. Opt. Soc. Am. A 12, 195 (1995)]. Consequently, when entrance and exit pupil sizes are equal, phase information is lost from the double-pass images. Second, we show that in double-pass measurements the eye behaves like a reversible optical system. That is, when entrance and exit pupils are equal, the double-pass image results from two passes through an optical system having a transfer function that is the same in both directions. To test for reversibility in the living eye we have used a double pass apparatus with different exit and entrance pupil sizes (one of them small enough to consider the eye diffraction limited), so that the ingoing and the outgoing transfer functions are different. The measured image quality was unchanged when the pupils were interchanged, i.e., when the first-pass entrance pupil size becomes the second-pass exit pupil size, and vice versa. Third, the technique provides a means for inferring the complete optical transfer function of the eye, including the phase transfer function, and the shape of the point spread function. PMID- 7500216 TI - Statistical distribution of foveal transverse chromatic aberration, pupil centration, and angle psi in a population of young adult eyes. AB - Subjective transverse chromatic aberration (sTCA) manifest at the fovea was determined for a population of 85 young adults (19-38 years old) by means of a two-dimensional, two-color, vernier alignment technique. The statistical distribution of sTCA was well fitted by a bivariate Gaussian function with mean values that were not significantly different from zero in either the horizontal or the vertical direction. We conclude from this result that a hypothetical, average eye representing the population mean of human eyes with medium-sized pupils is free of foveal sTCA. However, the absolute magnitude of sTCA for any given individual was often significantly greater than zero and ranged from 0.05 to 2.67 arcmin for the red and the blue lights of a computer monitor (mean wavelengths, 605 and 497 nm, respectively). The statistical distribution of the absolute magnitude of sTCA was well described by a Rayleigh probability distribution with a mean of 0.8 arcmin. A simple device useful for population screening in a clinical setting was also tested and gave concordant results. Assuming that sTCA at the fovea is due to decentering of the pupil with respect to the visual axis, we infer from these results that the pupil is, on average, well centered in human eyes. The average magnitude of pupil decentration in individual eyes is less than 0.5 mm, which corresponds to psi = 3 deg for the angle between the achromatic and the visual axes of the eye. PMID- 7500218 TI - Visual assessment of variable-resolution imagery. AB - A technique is described for producing variable-resolution images whose spatial detail decreases as a function of distance from their centers. These images can be matched in some sense to the normal spatial inhomogeneities of the human visual system, as well as to various abnormalities in spatial discrimination. A set of images was generated with a series of linear distortion functions whose low-pass characteristics differed at both the center and the periphery of the image as well as across the image. A forced-choice procedure was used to determine which test images were indistinguishable from unprocessed versions of themselves. Certain of the threshold distortion functions are compared with eccentricity scaling functions that have been used by others to characterize various aspects of peripheral vision. Finally, the concept of locally band limited spaces is discussed, and an efficient sampling technique based on the concept is described. This technique can be used to generate an image that, under certain conditions, is visually equivalent to an otherwise identical image containing significantly more information. PMID- 7500220 TI - Continuous assessment of perceptual image quality. AB - The study addresses whether subjects are able to assess the perceived quality of an image sequence continuously. To this end, a new method for assessing time varying perceptual image quality is presented by which subjects continuously indicate the perceived strength of image quality by moving a slider along a graphical scale. The slider's position on this scale is sampled every second. In this way, temporal variations in quality can be monitored quantitatively, and a means is provided by which differences between, for example, alternative transmission systems can be analyzed in an informative way. The usability of this method is illustrated by an experiment in which, for a period of 815 s, subjects assessed the quality of still pictures comprising time-varying degrees of sharpness. PMID- 7500219 TI - Pedestal effect in visual motion discrimination. AB - Many sensory discriminations, including the discrimination of speed, obey Weber's law and thus become more difficult as the stimuli get larger. Using one-jump apparent motion stimuli, we find that the opposite can occur: displacement discrimination improves with larger jumps. This pedestal effect occurs for small jumps near and below the detection threshold. Finding a pedestal effect in motion discrimination confirms a speed energy model developed in previous experiments on the detection of jump pairs, since the pedestal effect will be observed if the visual system detects the energy of the speed waveform. Once the size of the jumps becomes large enough, the discriminability declines, indicating masking. Masking is just the detectability counterpart of Weber's law; it is not predicted from energy detection. The pedestal effect shows the presence of a squaring nonlinearity for small speed signals, and masking indicates linear transduction for large signals. A half-wave rectifier, when presented with Gaussian noise, behaves this way. The speed energy model can be seen as an approximation, valid for small signals, to a model that includes half-wave rectification. PMID- 7500221 TI - Estimation of variance distribution in three-dimensional reconstruction. I. Theory. AB - A theory is developed for estimating the three-dimensional (3-D) variance of a 3 D image reconstructed from projections by weighted backprojection. The theory is applicable for any data-collection schemes that produce partially redundant sampling of the angular space. The particular data collection considered here, the single-exposure random-conical scheme, is used for the reconstruction of macromolecules in electron microscopy. In this context, the purpose of the 3-D variance estimation is to detect and localize the conformational variability, to assess the significance of structural differences between two experimentally related 3-D images, and to assess the significance of local features in a 3-D image. The 3-D variance estimate of each reconstruction voxel is obtained by (i) the comparison of closest points on Fourier sections associated with difference projections, (ii) the comparison of neighbor projections in real space, or (iii) the comparison of projections with reprojections of the reconstruction. PMID- 7500222 TI - Estimation of variance distribution in three-dimensional reconstruction. II. Applications. AB - A previously developed theory of three-dimensional (3-D) variance estimation [J. Opt. Soc. Am. A 12, 2615-2627 (1995)] is applied to the structural study of a hemocyanin-Fab complex with the electron microscope. The precise locations of structurally variable regions of the macromolecule are determined from the 3-D variance maps. The structural differences among different classes of the macromolecular complex are assessed by the use of the statistical t-test, and the 3-D antibody binding sites are revealed. From a model analysis, a rule is demonstrated for visually identifying a 3-D conformational change by the inspection of the 3-D variance map. Our analysis lays the foundation for numerous practical applications of variance estimation in the 3-D imaging of macromolecules. PMID- 7500224 TI - Sources of alcohol and drug information among Israeli urban adolescents. AB - This article describes a study which investigated sources of alcohol and drug information among Israeli urban adolescents in the north of Israel during the winter of 1993. Data were obtained from a sample of 1,346 students (50.9% males, 49.1% females), who were asked to indicate the amount of information (none, little, much) they had received from ten sources about eight categories of drugs. Among the findings: television is the primary source of information for all drugs, except inhalants, for which newspapers/magazines are the main source. Newspapers/magazines are of secondary importance for the rest of the seven categories of drugs. Teenagers are less likely to receive information from physicians/nurses and relatives than from other sources. Information based on personal experience is prominent concerning alcohol, cigarettes, inhalants and hashish/marijuana, and it prevails more among males than among females (p < .01). Females use school teachers as a source of information in all drug categories more than males (p < .01). There are differences in using various information sources among grades and types of places of living. Information based on personal experience with alcohol is notably correlated with that with cigarettes, whereas information based on personal experience with opiates is correlated with that with stimulants, hallucinogens and hashish/marijuana (p < .001). The mean number of sources of information used by the respondents is the highest concerning alcohol and the lowest concerning depressants. Implications for prevention are discussed. PMID- 7500223 TI - Continuation high schools: youth at risk for drug abuse. AB - Students at alternative high schools may be at substantial risk for drug abuse. The present article provides a general overview of the drug use-related context of continuation high schools in southern California. A total of 144 students and ninety-six staff were interviewed from twenty continuation high schools. The interview data revealed that continuation school students show high levels of substance use. However, only 20 percent of the students report that they received any drug abuse prevention programming. Also, students at continuation high schools aspired to a productive life after high school including continued education. Thus, these youth may still be amenable to preventive educational interventions which deter them from drug use and help them to fulfill their future goals. PMID- 7500225 TI - Beyond the schools: strategies for implementing successful drug prevention programs in community youth-serving organizations. AB - This article discusses why community-based youth-serving organizations are natural settings for drug prevention programming. Based on a three-year study of a school-based drug prevention program adapted to the Boys & Girls Club setting and serving high-risk youths, we identify organizational impediments to implementing a structured prevention program in youth organizations. We also present some strategies the clubs used to achieve success in a nonschool setting where program participation is voluntary. Strategies include: 1) employing a team approach and making program institutionalization a goal; 2) choosing the "right" prevention program leader; 3) creating a special prosocial bonding group for program youths; 4) involving program graduates as recruiters and positive role models; and 5) developing community support. Drug prevention programming in youth serving organizations can reinforce, supplement, and enhance school-based prevention efforts, thus providing communities with important drug prevention alternatives that go beyond the schools. PMID- 7500226 TI - Assessing organizational effectiveness in higher education drug prevention consortia. AB - Eighty-three consortia of institutions of higher education, organized under funding from the Fund for the Improvement of Post-Secondary Education (FIPSE) Drug Prevention Programs of the Department of Education, were surveyed to measure organizational effectiveness. Generalized satisfaction with the functioning of the consortia was related to the number of active members, the average miles traveled to meetings, satisfaction with performance of task functions, members' roles, the level of trust among members and the level of creativity and innovation in problem-solving. Satisfaction with goal attainment was significantly related to the presence of at least one "internal" goal for the consortium. PMID- 7500227 TI - A systematic review of parent-oriented programs to prevent children's use of alcohol and other drugs. AB - The purpose of this review was to analyze systematically the characteristics of twenty-two instructor-led parent-oriented programs. An objective of each of these programs was to prepare parents to prevent their children's alcohol and other drug use. To conduct this program review, the author developed 120 criteria for analyzing and reviewing the characteristics of the programs. The criteria were grouped under five main topics or components: 1) general characteristics, 2) instructional characteristics, 3) skills addressed, 4) generalization methods, and 5) evaluation. The author then trained two uninformed reviewers to review with reliability each program. The results of the reviews are summarized according to program characteristics. Recommendations are based upon the review results. These results and recommendations can help prospective users make an informed decision before they adopt or invest in a program. PMID- 7500228 TI - The cognitive structure underlying heroin-injecting behaviour. AB - An important objective in the field of drug education is the development of successful intervention programs to deter or reduce use. A vital step in this procedure is the identification of the critical elements that will induce and maintain behavior change. The purpose of this article is to demonstrate how the Theory of Reasoned Action can be used to identify these factors. Data were gathered from a sample of current heroin injectors and the cognitive structure underlying drug use investigated. The findings are discussed with regard to intervention programs aimed at modifying or reducing drug injecting behavior. PMID- 7500229 TI - Drug education in tune. AB - America's impassioned involvement with music has found its way into the schools. A great song is complete in itself and needs nothing more to enhance it. The nose thumbing spirit of Vietnam protesters in the seventies was reflected in the music of American songwriters of that period which included John Prine. Written in the traditional folk music style, John Prine's "Sam Stone" follows the storyline of a Vietnam veteran addicted to heroin and his tragic death. This article examines the heroin-related lyricism and music of "Sam Stone" as a teaching method in drug education. PMID- 7500230 TI - Case reports in pediatric psychology: uses and guidelines for authors and reviewers. AB - Described uses of case reports in pediatric psychology and guidelines for authors and reviewers. Case reports have several important functions: to document the efficacy of new treatment applications, describe interesting new clinical phenomena, to develop hypotheses, to illustrate methodological issues, difficult diagnostic issues, or novel treatment approaches, and to identify unmet clinical and research needs. Authors should carefully document the scientific purpose and clarify the new information contributed by their reports, provide evidence for their conclusions, and articulate clinical, theoretical, and/or research implications. PMID- 7500231 TI - Late psychosocial consequences for parents of children who survived cancer. AB - Investigated late psychosocial sequelae in 133 parents of children who survived cancer, using questionnaires developed to measure the specific impact of the disease. Childhood cancer had distinct and persistent late psychosocial effects on parents of survivors. Uncertainty and loneliness were the most reported problems. Demographic and situational characteristics such as being a mother, low SES, no religious affiliation, chronic disease in a family member other than the child surviving cancer, and concurrent stresses increased the risk of reporting late problems. Treatment itself had little or no effect on reported problems. However, when treatment led to long-term sequelae in the child, a significant and specific effect on parental problems was observed. No decline of problems over time was found, which has implications for patient care. PMID- 7500232 TI - Determinants of responsiveness in mothers of children with intraventricular hemorrhage. AB - Explored relationships between maternal responsiveness and parenting stress, family, medical, and demographic factors in mothers of children born prematurely at very low birth weights with intraventricular hemorrhage. Mothers of 82 2-year olds completed self-report questionnaires and were observed interacting with their children in free play during a yearly assessment. Path analysis (LISREL) revealed that lower maternal responsiveness, measured by the Parent/Caregiver Involvement Scale, was associated with greater demographic disadvantage and with high parent-related stress on selected subscales of the Parenting Stress Index. Child-related parenting stress was not associated with responsiveness, and severity of the child's medical condition at birth was weakly related. Results are discussed in the context of this unique population, and relevant research and measurement issues. PMID- 7500233 TI - Family coping with pediatric leukemia: ten years after treatment. AB - As part of a longitudinal study of family coping with pediatric leukemia, 28 former patients (16 male; 12 female; M age = 19.1 years) and their parents (23 mothers; 12 fathers) participated in a follow-up study at 10 years posttreatment. Measures included the Current Adjustment Rating Scale, the Brief Symptom Inventory, the Ways of Coping Scale, the Family Coping Scale, and a semistructured interview. Long-term survivors and their parents continued to be well-adjusted to life posttreatment. Coping and perceived adjustment in long-term survivors were positively related to socioeconomic status and mother's coping and negatively related to academic problems. A strong bidirectional relationship was found between survivors' and mother's adjustment. Coping strategies were variable and not significantly correlated with coping adequacy or adjustment. PMID- 7500234 TI - Perceived role restriction and adjustment of mothers of children with chronic physical disability. AB - Examined the relationship between maternal perceived role restriction, the extent to which a mother feels unable to pursue her own interests due to responsibilities with raising a child with a chronic physical condition, and psychosocial adjustment in 50 mothers with children 6-11 years old who had a chronic physical disability. Perceived role restriction accounted for a significant increment in the variation in adjustment beyond that contributed by objective indices of the child's disability. Neither these objective parameters of the child's disability nor his or her level of problem behaviors predicted perceptions of role restriction. However, perception of social support did. The potential role of intrapersonal processes was discussed as a neglected component of a multidimensional model of adjustment in mothers of children with a chronic physical condition. PMID- 7500236 TI - Otitis media in early childhood and cognitive, academic, and behavior outcomes at 12 years of age. AB - Examined the association between otitis media with effusion (OME) during the first 3 years of life and cognitive and academic performance and behavioral outcomes at 12 years of age in 56 socioeconomically disadvantaged children attending a research childcare program. OME history was prospectively documented from birth through 3 years during well and illness periods. Standardized tests of intelligence and academic achievement and measures of behavior were administered when children were 12 years of age. OME during early childhood was not related to intellectual performance, academic achievement, behavior, or attention. Results do not support a long-term relationship between a history of OME and later developmental outcomes. Generalizations of these findings to other populations should be done cautiously. PMID- 7500235 TI - Pediatrician's communication style: relationship to parent's perceptions and behaviors. AB - Investigated physician-parent communication styles and the effects of concordance between parent's desired communication styles and the communication style exhibited by physicians. Subjects were 107 parents of children scheduled for an appointment with a pediatrician at a general medical clinic. Parents and physicians completed rating forms indicating the degree to which parents desired each of four communication styles (information giving, interpersonal sensitivity, partnership, and directing one's own treatment). Parents and physicians also rated the degree to which they believed the physician exhibited each of these four styles. Follow-up interviews with parents assessed the level of satisfaction with the visit, perception that parent's concerns had been addressed, and subsequent telephone calls to the physician. Results indicated that physicians underestimated the degree of interaction desired by the parents. Parent desires for particular communication styles were not predicted by characteristics of the parents. Interaction variables predicted parent perceptions and subsequent need for contact with the physician. PMID- 7500237 TI - Somatic symptoms in sexually abused children. AB - Contrasted a nonabused sample of 3 to 12-year-old children (n = 847) with a sexually abused sample (n = 252) with regards to somatic symptoms assessed via parent report, including the Child Behavior Checklist. Using ANCOVA, and controlling for family income, maternal education, age, and sex, the girls 3-6 years old who were abused and the boys 7-12 years old who were abused were reported to have the greatest number of somatic symptoms. Force and number of perpetrators were abuse-specific variables directly related to the number of somatic symptoms. Results suggest that for some children sexual abuse is related to an increased report of subjective somatic complaints. PMID- 7500238 TI - Misconceptions about AIDS among children who can identify the major routes of HIV transmission. AB - Described knowledge about HIV transmission among 1,048 third, fourth, fifth, and sixth graders from a multiethnic urban school district. Participants answered questions about behaviors leading to HIV transmission and about mechanisms of transmission. Children at all grade levels demonstrated a high level of recognition of the three primary routes of HIV transmission (through sex, through drug needle sharing, and in utero). However, children who knew about these routes also had many misconceptions about the mechanisms involved in acquiring HIV through these routes and about the ways HIV is not transmitted. Misconceptions decreased with grade and socioeconomic status (SES). Although there were no race/ethnicity differences in knowledge of actual routes and mechanisms of HIV transmission, there were more misconceptions about routes and mechanisms of HIV transmission among African American and Asian American children than among white children. PMID- 7500239 TI - A Comparative study of ePTFE periodontal membranes with and without decalcified freeze-dried bone allografts for the regeneration of interproximal intraosseous defects. AB - A comparative study which evaluated two treatment modalities for regeneration of interproximal periodontal lesions was conducted. Eleven pairs of advanced periodontal lesions in 11 human subjects (6 male and 5 female) were treated in a split mouth design with expanded polytetrafluoroethylene (ePTFE) interproximal membranes alone (control) in one lesion and ePTFE interproximal membranes in combination with decalcified freeze-dried bone (test) in the other lesion. At 1 year post-treatment, both treatment modalities revealed a significant increase in clinical attachment levels from baseline (2.0 +/- 1.37 mm test, 2.0 +/- 0.88 mm control) with no significant differences between the two modalities. When the amount of new bone formed in these lesions was assessed using re-entry procedures, a significant difference in new bone formation from baseline was found only for lesions treated with ePTFE alone (0.4 +/- 0.78 mm test, 1.3 +/- 0.96 mm control). It was concluded that at 1 year post-treatment, significant clinical attachment gains could be obtained by the use of ePTFE barriers with or without DFDBA. Statistically significant results in bone fill were only found when ePTFE barriers were used alone. PMID- 7500240 TI - Effect of instrument power setting during ultrasonic scaling upon treatment outcome. AB - Ultrasonic scalers may be operated at different power settings which may influence the final therapeutic result. The displacement amplitude of the scaling tip may affect scaling efficiency or the degree of root surface damage. This investigation aimed to determine whether there was any difference in periodontal healing, as assessed by standard clinical methods, when an ultrasonic scaler was operated at full or half power in patients suffering from mild to moderate adult periodontitis. Seventeen patients with chronic adult periodontitis were entered into the study and a split mouth design utilized. Quadrant allocation was randomized, 2 quadrants being treated at full power and 2 at half power. At baseline, probing attachment levels, bleeding and plaque indices (BI and PI) were recorded and oral hygiene instruction given. Two weeks later all measurements were repeated and further recordings made at 1, 3, and 6 months post-therapy. Statistical analysis demonstrated no differences between groups at baseline (P > 0.3) for all parameters. At the post-oral hygiene stage, there were significant attachment gains for both experimental groups (P < 0.04), but no differences were found between the groups (P > 0.6). At 6 months, mean attachment gains for the half power and full power groups over baseline measures were 0.5 +/- 0.5 mm and 0.4 +/- 0.3 mm, respectively (P < 0.002), and there were no significant differences between the groups (P > 0.5). The results show that the use of the half power setting was as effective as using the ultrasonic scaler at full power.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7500243 TI - Immunohistochemical study of gamma delta T cells in human gingival tissues. AB - The distribution and the density of gamma delta T cells in human gingival tissues were examined immunohistochemically in biopsy samples obtained from 20 subjects. Few gamma delta T cells were observed in gingival tissue free from inflammatory cell infiltration, but were found, albeit in low numbers, in association with inflammatory cell infiltration, especially T cells. This relationship with T cells was confirmed statistically. The ratios of gamma delta T cells to T cells in the epithelia and in the connective tissue were calculated in the sections in which more than 500 CD3-positive cells were identified. Seven of eight such epithelial specimens showed a ratio of less than 1% and one less than 2% (mean +/ SD; 0.8% +/- 0.4). In the connective tissue, 8 of 13 such specimens showed less than 1%, three less than 2%, one 3%, and one 7% (1.4% +/- 1.9). These results suggest that basically gamma delta T cells are not resident cells in the gingival epithelium such as comprise the first defense line against exogenous irritation. They may play some role in the pathogenesis of periodontal disease collaborating with alpha beta T cells in the inflammatory response. PMID- 7500242 TI - Role of saliva and salivary components as modulators of bleaching agent toxicity to human gingival fibroblasts in vitro. AB - Mild oxygenating agents generating H2O2 are used for effective at-home tooth bleaching, but can cause gingival ulcers in some patients. There are concerns about the possible pathological effects of relatively long-term exposure of oral tissues to bleaching agents. Previous work in our laboratory showed that a bleaching agent, which generates approximately 3% H2O2 from carbamide peroxide, was toxic to human gingival fibroblasts in vitro, but that the toxicity was abolished by treatment with the H2O2-destroying enzyme catalase. The purpose of the present study was to determine if whole saliva, the salivary enzyme lactoperoxidase (LP) (which, like catalase, removes H2O2), or salivary mucin protected fibroblasts from bleaching agent toxicity. The cells were exposed to 0.05% agent with or without saliva, LP, mucin or catalase (as a positive control based on our previous study) and assessed for effect on viability/morphology (by microscopic observation), proliferation (by [3H]-thymidine incorporation), and the production of fibronectin (FN) and type I collagen (by ELISA). While the bleaching agent at 0.05% caused cell death, the cells appeared viable and morphologically normal when treated with the bleaching agent and LP (> or = 0.1 microM), saliva LP, and catalase from agent inhibition of proliferation (P < or = 0.04) and FN production (P < or = 0.01). Mucin had statistically insignificant or no protective effect as assessed by the above parameters. Treatment with saliva, LP, mucin, and catalase gave complete or partial protection from agent-inhibition of collagen production (P < or = 0.04).(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7500241 TI - Interleukin-1 beta gene expression in human oral polymorphonuclear leukocytes. AB - Oral polymorphonuclear leukocytes (PMN) were obtained from 10 adult donors in good oral health using a method employing repeated mouth rinse collection. Interleukin-1 beta (IL-1 beta) mRNA was detected in freshly obtained cells by blot hybridization of total cellular RNA with a biotin labeled cDNA probe. Supernates from oral PMN placed in culture for 3 hours contained substantial amounts of IL-1 beta measured by ELISA. Significantly greater numbers of PMN and amounts of PMN-derived IL-1 beta were obtained from the same donors 2 hours subsequent to an oral sucrose challenge (3.23 x 10(6) vs. 1.57 x 10(6) mean PMN number, P = 0.004; 59.80 vs. 20.05 mean pg/ml IL-1 beta, P = 0.036, respectively). However, the elevated levels of IL-1 beta were due to the higher cell number rather than to increased production by individual cells. Stimulation of oral PMN with recombinant granulocyte-macrophage colony stimulating factor did not enhance their cytokine production. In most instances. IL-1 beta production by oral PMN was dramatically greater than that of their blood counterparts. These findings suggest that oral PMN are an important source of IL-1 beta, which plays a central role in oral immunity and inflammatory disease states. PMID- 7500244 TI - Gamma delta T lymphocytes in human periodontal disease tissue. AB - Gamma delta T cells have been implicated as playing a role in inflammatory and autoimmune conditions. In this study, a biotin-streptavidin peroxidase technique was used to determine the presence of T cells expressing alpha beta and gamma delta T cell receptors (TCR) in the inflammatory infiltrates of gingival tissue sections from gingivitis and adult periodontitis patients. The lesions were graded 1+ to 3+ according to the size of the infiltrate. alpha beta+ T cells predominated in all infiltrates with no differences in the mean percent (20 to 30%) according to size of lesion or to clinical status. A mean percent gamma delta T cells of 0.42 +/- 0.11 and 0.91 +/- 0.42 was demonstrated in 1+ infiltrates of gingivitis and adult periodontitis sections respectively. Although the mean percent gamma delta T cells increased in both gingivitis (2.09 +/- 0.54) and adult periodontitis sections (2.25 +/- 0.35) with increasing size of infiltrate, this was not statistically significant. However, when the mean proportion of gamma delta T cells of the total TCR bearing cells was determined, there was a significant 3 to 4 fold increase in adult periodontitis sections from 3.09 +/- 1.35 in 1+ lesions to 11.90 +/- 2.94 and 8.81 +/- 1.45 in 2+ and 3+ lesions respectively. A similar increase of the same magnitude occurred in gingivitis sections from 2.82 +/- 0.74 in 1+ lesions to 11.12 +/- 4.13 in 2+ lesions, but this was not significant (P = 0.055). There was no correlation between the increase in the proportion of gamma delta T cells and the T:B cell ratio or the CD4:CD8 ratio in individual lesions.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7500246 TI - Periodontal regeneration of human intrabony defects with titanium reinforced membranes. A controlled clinical trial. AB - The purpose of this controlled clinical trial was to compare the clinical efficacy of 3 treatment modalities in the treatment of deep interproximal intrabony defects. Forty-five (45) defects in 45 patients were randomly assigned to 1 of 3 treatment groups by blocking to prognostic variables. The test group was treated with titanium reinforced membranes positioned just apical to the cemento-enamel junction and the modified papilla preservation technique; the second group received conventional expanded polytetrafluoroethylene (ePTFE) barrier membranes applied at the alveolar crest; the third group was treated with an access flap procedure. The groups were well balanced with respect to all prognostic variables. During the 1-year observation period, patients were subjected to a stringent infection control program including: professional tooth cleaning every week for the first 6 weeks (all groups) and in the 4 weeks following membrane removal (guided tissue regeneration groups), then at monthly intervals (all groups). The results indicated that: 1) all treatment modalities resulted in clinical and statistically significant improvements in clinical attachment level (CAL) and probing depths (PD) at 1 year; 2) a significantly greater amount of CAL gain (P = 0.0003, analysis of variance) was observed in the test group (5.3 +/- 2.2 mm) with respect to both the ePTFE group (4.1 +/- 1.8 mm) and the flap control group 2.5 +/- 0.8; 3) in the test group the 1 year CAL (4.7 +/- 1.8 mm) was located more coronally than the baseline position of the interproximal alveolar crest (5.9 +/- 2 mm; P = 0.003, t test).(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7500247 TI - Primary extramedullary plasmacytoma in cyclosporine-induced gingival overgrowth. A case report. AB - An unusual solitary extramedullary plasmacytoma developed in the gingiva of a 28 year-old male renal transplant patient with cyclosporine (CyA)-induced gingival overgrowth. On initial presentation, there was no suspicion of a tumor within the lesion. One year later, however, the lesion had increased progressively in size, and its surface was ulcerated. At this time, the lesion was excised by gingivectomy, and histopathological examination revealed plasmacytoma. CyA was discontinued and the patient has been closely observed for 4 years with no recurrence of the gingival lesion. PMID- 7500248 TI - Infantile genetic agranulocytosis (Kostmann type). A case report. AB - Severe periodontitis in a patient with infantile genetic agranulocytosis (Kostmann syndrome) is presented. This rare syndrome is inherited as an autosomal recessive pattern and characterized by severe neutropenia. The periodontal status and treatment of the patient is described. It is concluded that periodontal therapy including scaling, root planning, soft tissue curettage, and the use of selected antimicrobial agents can be successful in this particular syndrome. PMID- 7500249 TI - Massive gingival enlargement and alveolar bone loss: report of two cases. AB - We present two cases of massive gingival enlargement and osteolysis of alveolar bone in a 30-year-old female and a 36-year-old male. The etiology could not be established in either case. Histologically, both lesions contained hyperplastic fibrous connective tissue and intense plasma cell infiltrates. Both patients responded well to extensive gingivectomy, extraction of all teeth, and alveoplasty. PMID- 7500245 TI - Hydrogen peroxide: a review of its use in dentistry. AB - Several dentifrices that contain hydrogen peroxide are currently being marketed. The increased use of bleaching agents containing (or generating) H2O2 prompted this review of the safety of H2O2 when used in oral hygiene. Daily exposure to the low levels of H2O2 present in dentifrices is much lower than that of bleaching agents that contain or produce high levels of H2O2 for an extended period of time. Hydrogen peroxide has been used in dentistry alone or in combination with salts for over 70 years. Studies in which 3% H2O2 or less were used daily for up to 6 years showed occasional transitory irritant effects only in a small number of subjects with preexisting ulceration, or when high levels of salt solutions were concurrently administered. In contrast, bleaching agents that employ or generate high levels of H2O2 or organic peroxides can produce localized oral toxicity following sustained exposure if mishandled. Potential health concerns related to prolonged hydrogen peroxide use have been raised, based on animal studies. From a single study using the hamster cheek pouch model, 30% H2O2 was referred to as a cocarcinogen in the oral mucosa. This (and later) studies have shown that at 3% or less, no cocarcinogenic activity or adverse effects were observed in the hamster cheek pouch following lengthy exposure to H2O2. In patients, prolonged use of hydrogen peroxide decreased plaque and gingivitis indices. However, therapeutic delivery of H2O2 to prevent periodontal disease required mechanical access to subgingival pockets. Furthermore, wound healing following gingival surgery was enhanced due to the antimicrobial effects of topically administered hydrogen peroxide. For most subjects, beneficial effects were seen with H2O2 levels above 1%. PMID- 7500250 TI - Axonal transport of fluorescent carbocyanine dyes allows mapping of peripheral nerve territories in gingiva. AB - Sensory innervation of gingival tissue can cause neurogenic inflammation that depends on the extent of the branching area of the peripheral nerve fibers. We designed the present study to determine whether single trigeminal axons branch to both the buccal and palatal gingiva of maxillary molars of adult rats. Accumulation via retrograde transport of DiI (red) or DiA (green) fluorescent carbocyanine dyes in neurons of trigeminal ganglia was evaluated 7 days after applying one dye to the buccal sulcus and the other to the palatal sulcus. Both dyes were absorbed through the junctional epithelium, and the two sites each labeled similar numbers and sizes of neurons in the lateral zone of the maxillary division (44% from buccal and 46% from palatal gingiva). Double-labeled neurons had the same size (32.5 +/- 6.70 microns, mean circumference +/- S.D.) and location as single-labeled neurons, and they were 9% of the total. This study shows that exogenous dyes can diffuse into mucosa and thereby allow in vivo mapping of sensory nerve branching patterns to several intact tissues per animal. We found that 9% of the labeled cells extended to both the buccal and palatal gingiva. Thus, inflammation that spreads from one gingival region to the other could have a neurogenic mechanism involving trigeminal sensory neurons that extend their peripheral branches to innervate both buccal and palatal gingiva of adult rat molars. PMID- 7500252 TI - Nursing's response to health care reform: an update. PMID- 7500253 TI - Management of nausea and vomiting in children. AB - When children experience nausea and vomiting (N/V) as side effects of anesthesia or chemotherapy, a major role for nurses is to promote comfort. In addition to the discomfort of N/V in children, other detrimental effects include dehydration, weight loss, fluid and electrolyte imbalance, and emotional distress. This article describes the physiological pathways through which the vomiting center is stimulated to cause nausea and vomiting and identifies appropriate interventions for blocking these pathways using both pharmacological and nonpharmacological means. Medications used to control nausea and vomiting, including phenothiazines, substituted benzamide, corticosteroids, benzodiazepines, antihistamines, and serotonin antagonists, are discussed. Nonpharmacological interventions that are described include reducing anxiety, music therapy, hypnosis, progressive muscle relaxation, dietary modifications, and acupressure. PMID- 7500251 TI - Human demineralized freeze-dried bone: inadequate induced bone formation in athymic mice. A preliminary report. AB - The purpose of this study was to test the osteoinductive properties of demineralized freeze-dried bone (DFDBA) randomly purchased from four commercial bone banks. Twenty-five (25) milligrams of bone from each of the banks was implanted into the hindquarter muscles of athymic mice. Two samples from each of the banks were compared with samples from the other banks. A total of 16 implants were grafted into 8 mice. Two additional mice served as controls. One mouse received an implantation of deactived human cortical bone matrix (DBM) (negative control). The other mouse received an implant of human bone morphogenetic protein/non-collagenous proteins (hBMP/NCP) infused to surface demineralized human cortical bone (positive control). At 21 days the mice were killed, the hindquarters were photographed, and the tissues were prepared for histologic evaluation. Of the 16 commercial DFDBA implants, 12 were available for histologic evaluation. There was no radiographic evidence of bone formation for the DFDBA implanted mice or the DBM implants. Small bone ossicles were scarcely visible in the hindquarters of the mouse which received the hBMP/NCP infused bone. Histomorphometric analysis was used to determine the percentage of new and dead bone. The bone was measured in pixels. The predominant histologic feature of the DFDBA implants was non-vital bone chips with minimal amounts of new bone. The average amount of non-vital bone ranged from 78.4% to 92.5%. There was no evidence of bone formation for the DBM implants. The average amount of bone for the mouse which received hBMP/NCP was 96%. The results of this pilot study indicate that commercially-available DFDBA induced clinically insignificant amounts of bone.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7500254 TI - Mastery motivation and cognitive development in 4-year-old children born at various degrees of medical risk. AB - Although technology has enabled premature infants to survive, longitudinal assessments have primarily focused on IQ outcomes. The purpose of this study was to establish the usefulness of mastery assessment in establishing developmental outcome. Mastery motivation and cognition were examined in 167 preschool children born at various degrees of medical risk. Mastery motivation was measured by a videotaped toy sequence designed to tap sensorimotor practice, effect production, and problem solving. Patterns of correlations indicate that the relationship between mastery and IQ were complementary and not redundant. Three regression models showed that significant interactions between perinatal status and mastery motivation behaviors confirm mastery assessment worthy of consideration in biologically high-risk children. PMID- 7500255 TI - Fetal alcohol syndrome from infancy through childhood: a review of the literature. AB - Because fetal alcohol syndrome (FAS), or fetal alcohol effects (FAE), is being diagnosed in growing numbers of children, there is more interest in their common characteristics and how they change throughout childhood. Few longitudinal studies that describe the common childhood course of FAS address appropriate interventions. Some literature exists that describes follow-up and support services needed by these children and their families. This article examines the common childhood course of FAS/FAE and the interventions and services needed to promote positive child and family health. PMID- 7500256 TI - A cost analysis of a case management system for infants with chronic illnesses and developmental disabilities. AB - Service coordination has long been a documented need of children with disabilities. The purpose of this study was to examine the costs associated with providing a comprehensive system of service coordination for hospitalized infants and toddlers with special health care needs and their families. Coordination costs were evaluated across seven functions including (a) determining eligibility for services, (b) identifying and arranging evaluations, (c) providing support to families, (d) making referrals to outside agencies, (e) exchanging information among service providers and families, (f) maintaining follow-up contact, and (g) determining discharge from the program. Results indicated that the service coordination function of providing family support was the most time-consuming task area, followed by the functions of exchanging information and maintaining follow-up contact. Costs also varied with the medical diagnosis and the child's age. Consistent with this variability, the diagnostic category and/or possibly the length of hospitalization was a better correlate of total cost of service coordination per child than was the number of months served. The complexity of the family's social and financial situation also appeared to be related to cost per month of service. PMID- 7500257 TI - Perceived stressors and coping strategies of parents who have children with developmental disabilities: a comparison of mothers with fathers. AB - The purposes of this study were to describe perceived stressors and coping strategies of parents who have children with developmental disabilities and to examine similarities and differences of mothers and fathers in their perceptions of stressors and coping. A sample of 203 parents of children with developmentally disabilities, ages two through five years, participated in the study. Hymovich's Parent Perception Inventory: Concerns (Hymovich, 1988) and the Ways of Coping Questionnaire (Folkmann & Lazarus, 1988) were used for data collection. Stressors and coping strategies were identified. The stressor reported by the largest number of both mothers and fathers was concern about the child's future. There were fewer differences between mothers and fathers in reporting of stressors than in reporting coping strategies. The importance of recognizing parental individuality in family coping is addressed within the context of implications for nursing practice and research. PMID- 7500258 TI - Exploring the concept of health among adolescents with diabetes using photography. AB - The purpose of this study was to explore and describe the concept of health among adolescents with diabetes. Photography was used to facilitate communication. Participants were given Polaroid cameras with instructions to take pictures illustrating health. They were then asked to explain in writing how their pictures demonstrated the concept. The participants produced 46 pictures and 53 descriptive expressions, which were analyzed for content. Health was described as energy, positive feelings, absence of illness, strength, activity, fitness, mental ability, and relationships. There were also descriptions of what one does to be healthy, which included eating nutritious food, exercising, engaging in diabetes-related care, sleeping, studying, relaxing, maintaining hygiene, and obtaining shelter. PMID- 7500259 TI - Ten ways to enhance the nursing profession's lobbying efforts with government. PMID- 7500260 TI - Iontophoresis for pain management. PMID- 7500261 TI - Children in Jordan: what is their health status and how do they perceive nurses? PMID- 7500262 TI - [Assessment of right ventricular function by magnetic resonance imaging of old myocardial infarction]. AB - The usefulness of magnetic resonance imaging (MRI) for estimating right ventricular function and the influence of left ventricular dysfunction on the performance of the right ventricle were assessed in 43 patients with chronic myocardial infarction (MI) and 14 control subjects (N) using electrocardiography gated MRI and cardiac catheterization. Patients with MI were divided into three groups according to the location of the coronary lesions; 22 patients with left coronary artery lesion (LCA group), 13 with right coronary artery lesion (RCA group), and 8 with both left and right coronary artery lesions (L+R group). The right ventricular end-diastolic volume (RVEDV) and right ventricular ejection fraction (RVEF) were measured by Simpson's rule algorithm on transverse images of the right ventricle obtained at the end-systolic and end-diastolic phases. In 34 of the 43 patients, the same parameters of right ventricular function were calculated by the thermodilution method using a Swan-Ganz catheter with rapid response thermistor. Left ventricular ejection fraction (LVEF) and end-diastolic volume were determined from left ventriculography. The intraobserver reproducibility (11 cases, r = 0.97) and interobserver reproducibility (11 cases, r = 0.92) of RVEF measured by MRI were excellent. The RVEF and RVEDV determined from MRI were significantly correlated with those from the thermodilution method (RVEF: r = 0.56, RVEDV: r = 0.52). There was no difference in right ventricular end-diastolic volume index in any patient group. The RVEF was decreased in the L+R (41.0 +/- 4.5%, p < 0.01) and RCA (45.9 +/- 6.6%) groups, but there was no difference between the LCA (50.6 +/- 6.6%) and N (48.9 +/- 4.3%) groups.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7500263 TI - [Acute myocardial infarction during sport]. AB - Thirty patients with acute myocardial infarction which occurred during sport were investigated to identify the type of sport, prodromata, situations at the onset of disease, habit of exercise, preceding medical evaluation, coronary risk factors, and coronary angiographic findings. Infarction occurred during golf in 12 patients, bowling in 4, gateball in 4, jogging or running in 5, baseball in 2, and tennis or table tennis in 3. The majority of the patients were playing ball games. Twenty-seven patients were men (90%) and 3 were women (10%). All patients had played the same kind of sport for several years. Twenty-four patients had one or more coronary risk factors, and especially 18 patients smoked cigarettes. Nine patients had experienced anterior chest pain but only two patients had received medical evaluation. Coronary angiography was performed in 25 patients (83.3%), revealing single-vessel disease in 14, two-vessel disease in 6, three-vessel disease in 4, and disease of all left main coronary trunks in 1. The acute episode of infarction occurred mainly in spring or fall. Many patients with acute myocardial infarction occurring during sport participate in sports of low or moderate dynamic and low static exercises which are generally regarded safe. Many patients had enjoyed their sports regularly for a long time. Though many patients had coronary risk factors, only a few had received a medical check before their heart attack. PMID- 7500264 TI - [Diagnosis and prognosis of elderly patients with coronary artery disease: assessment with dipyridamole thallium imaging]. AB - Noninvasive evaluation of coronary artery disease is difficult in elderly patients because of their limited exercise-capacity. The diagnostic and prognostic value of dipyridamole perfusion scintigraphy was assessed in 147 patients aged 65 years and older. All patients underwent coronary angiography. Initially, 32 patients had percutaneous transluminal coronary angioplasty (PTCA) or coronary artery bypass grafting (CABG). The other 115 patients were first treated medically and followed for a mean of 29 +/- 22 months. Nine patients (7.9%) had cardiac events including cardiac death and coronary intervention (PTCA or CABG) during the follow-up. Dipyridamole perfusion scintigraphy was performed safely in all patients, whereas treadmill exercise testing could not be adequately performed in 24 patients, 18 of whom had multivessel disease. Multiple regression analysis showed that: fixed defect and reversible defect were powerful detectors of coronary lesions (p = 0.0001, p = 0.0027, respectively), all patients with fixed disease and 94% of patients with only reversible defect had significant coronary lesion; Diffuse slow washout and ST depression were statistically significant for detecting multivessel coronary lesions in patients with fixed disease (p = 0.0001, p = 0.017, respectively), the sensitivity and specificity of diffuse slow washout and/or ST depression for detecting multivessel coronary lesions ware 85% and 74%, respectively, and ST depression was statistically significant for detecting multivessel coronary lesions (p = 0.0002) in patients with only reversible defect, the sensitivity and specificity of ST depression were 88% and 64%, respectively. Cox survival analysis identified diffuse slow washout as the best predictor of future cardiac events among the scintigraphic variables.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7500265 TI - [Initial and short-term results of percutaneous transluminal coronary angioplasty in patients over 70 years old: recent improvements in effectiveness]. AB - Elderly patients have a higher incidence of procedure-related complications and recurrent angina after percutaneous transluminal coronary angioplasty (PTCA). However, early success rates seem to have improved. To determine whether outcome of PTCA in the elderly has changed recently, we compared initial results and short-term outcome in patients over 70 years old in two groups; 64 patients with 70 PTCA lesions who underwent first elective PTCA in 1982-1989 (early group) and 140 patients with 153 PTCA lesions in 1990-1993 (late group). The mean age of the late group was significantly older than that of the early group (74 +/- 3 vs 73 +/- 2 years, p < 0.01). The two groups had similar clinical characteristics including sex, prior myocardial infarction and multivessel disease. The overall success rate of PTCA was significantly higher in the late group than in the early group (92.8% vs 82.8%, p < 0.05). The rate of abrupt closure was similar in the two groups (2.9% vs 2.6%). Major complications (procedural myocardial infarction, emergency bypass surgery, hospital death) occurred in 4.7% of the early group and in 3.6% of the late group. Angiographic restenosis occurred in 41.5% of the early group and in 33.6% of the late group. Cardiac events (cardiac death and/or myocardial infarction) occurred in 6.6% of the early group and in 1.5% of the late group (p = 0.06). The initial success rate of PTCA has improved in the elderly as well as younger patients (< 70; 92.8% vs 91.6%). We conclude that PTCA can be performed with high initial success and satisfactory short-term outcome in patients over 70 years old. PMID- 7500266 TI - [Transesophageal echocardiographic study on the mechanisms of mitral regurgitation in hypertrophic cardiomyopathy: comparison with sigmoid septum]. AB - Transesophageal echocardiography was performed to elucidate the mechanisms of mitral regurgitation (MR) in 40 patients with hypertrophic cardiomyopathy with asymmetric septal hypertrophy, 15 obstructive and 25 nonobstructive, and the organic changes of the mitral leaflet were compared to those of 30 patients with sigmoid interventricular septum. Thirty subjects without cardiac diseases served as the control group. Transthoracic and transesophageal echocardiography were performed in all subjects to measure the following: left ventricular dimension, interventricular septal thickness and peak velocity at the left ventricular outflow tract by transthoracic echocardiography; the lengths and the thicknesses of the rough zone of the anterior and posterior mitral leaflets at mid-diastole and the distance between the tip of the posterior papillary muscle and the anterior mitral annulus by transesophageal echocardiography. The presence of systolic anterior motion of the mitral complex, contact between the anterior mitral leaflet and the interventricular septum during diastole, and the occurrence of mitral valve prolapse (MVP) were also investigated. The maximum area and timing of MR during systole was measured by M-mode color Doppler technique. The following results were obtained. 1. MR was observed in 35 (88%) of the 40 patients with hypertrophic cardiomyopathy. The maximum regurgitant area in the obstructive group was significantly greater than in the other groups, and the regurgitation was frequently pansystolic. 2. The lengths of both mitral leaflets at mid-diastole were significantly greater in both groups with hypertrophic cardiomyopathy than in the other groups.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 7500267 TI - [Treatment of refractory hypoxemia due to right-to-left interatrial shunt complicating right ventricular infarction: successful short-term percutaneous catheter closure of the patent foramen ovale]. AB - Refractory hypoxemia as a complication of right ventricular infarction is rare. A 74-year-old man developed persistent hypoxemia due to right-to-left interatrial shunting through a patent foramen ovale during the acute phase of right ventricular infarction. Short-term percutaneous transvenous closure of the patent foramen ovale with a large balloon occlusion catheter was life-saving. PMID- 7500268 TI - [Identical male twins showing progression from hypertrophic cardiomyopathy to dilated cardiomyopathy-like features]. AB - Twenty-three-year-old identical male twins with hypertrophic cardiomyopathy which progressed into the dilated phase are reported. The younger brothers first presented at age 16 with an abnormal electrocardiogram. Hypertrophic nonobstructive cardiomyopathy with an asymmetric septal hypertrophy was diagnosed. He was treated with beta-blocker, but he stopped taking the drug as he had no symptoms at that time. He presented again at age 21 years with symptoms of apparent congestive heart failure. Echocardiography showed marked dilatation of the left ventricle with thin wall which was compatible with dilated cardiomyopathy. The elder brother presented with an initial echocardiogram showing systolic anterior movement of the mitral valve without asymmetric septal hypertrophy. He presented again with his brother aged 21 years when his echocardiogram showed slight dilatation of the left ventricle, although he did not complain of cardiac symptoms. These identical twins are the first reported cases of hypertrophic cardiomyopathy progressing to the deteriorated dilated phase. PMID- 7500269 TI - Physical model for lesion formation in the presence of low levels of solution fluoride. AB - A quantitative physical model is presented for the formation of subsurface carious lesions in the presence of low levels of solution fluoride. Calculations using independently determined model parameters are in agreement with mineral density profiles measured in bovine enamel lesions. The proposed mechanism is controlled by fluoride in the following way: as fluoride diffuses into enamel, it is rapidly adsorbed to enamel crystallites, resulting in very low microenvironmental fluoride concentrations, so long as the crystals are not saturated with respect to fluoride adsorption. The result of this saturable adsorption is a widening band of fluoride-saturated crystals near the surface, beneath which the microenvironmental fluoride concentrations are negligible. In the saturated band, the microenvironmental fluoride concentration in the pore solution is high enough to suppress dissolution, while in the deeper, relatively fluoride free region, dissolution can occur. In addition to predicting observed mineral density profiles, the model also predicts the demarcation in solution conditions between the regime where subsurface lesion formation occurs and that where the dissolution pattern is that of surface erosion; and the lack of insensitivity of dissolution rate to hydrodynamics in the presence of low levels of fluoride, as contrasted to the square root of stirring rate dependency observed in the absence of fluoride. PMID- 7500272 TI - A robotic dissolution system with on-line fiber-optic UV analysis. AB - An automated, robotic system has been developed with on-line UV fiber optics for real time dissolution analysis of solid dosage forms. The system is comprised of "off-the-shelf" hardware including a UV-Vis diode array spectrophotometer, fiber optic coupler, immersion probe, robot, and dissolution apparatus. The software system is modular with the functionalities of control, data acquisition, and spectral analysis separated into three Windows applications with communications performed via Dynamic Data Exchange (DDE). The fiber optic spectrophotometer collects a full spectrum over the range of 190-810 nm. Single wavelength UV analysis is performed on dosage forms in six dissolution vessels. The robotic system automates all facets of the analyses: measuring, degassing, and dispensing of the media; thermostating the media to physiologic temperature; dropping the dosage forms into the vessels; immersing the fiber optic probe at the appropriate time intervals; initiating the data acquisition, analyses, and reporting; and emptying and washing of the vessels prior to the next automated run. As a representative dosage form, 10 mg active tablets were selected and analyzed by this method. This fiber optic system has significantly improved the throughput of the robotic systems by eliminating the need for time consuming off-line HPLC or UV analyses. In addition, with the exception of system calibration, it is no longer necessary for laboratory personnel to come in contact with samples. PMID- 7500273 TI - Iontophoretic delivery of oligonucleotides across full thickness hairless mouse skin. AB - In recent years there has been an increased interest in the use of oligonucleotides as therapeutic agents. Oligonucleotide therapeutics may have significant potential over traditional drugs due to their high degree of specificity and increased affinity. The major drawbacks to the use of oligonucleotide therapeutics are the problems associated with their delivery and their relative instability in serum. The serum instability problem has been partially overcome through the use of oligonucleotides with modified backbones. Transdermal electrotransport may be used to overcome the problems associated with delivery. Here we report the use of transdermal electrotransport in the delivery of oligonucleotides across hairless mouse skin. The effects of pH, salt concentration, current density, and oligonucleotide concentration, structure, and length have been investigated. PMID- 7500271 TI - Influence of physicochemical parameters and perfusate flow rate on the distribution of solutes in the isolated perfused rat hindlimb determined by the impulse-response technique. AB - The relationship between solute distribution, physicochemical properties, and tissue physiology was determined by the impulse-response (IR) technique and statistical moment analysis in the isolated perfused rat hindlimb. The concentration of bovine serum albumin (BSA; 2, 4.7, and 7%, w/v), perfusate flow rate (4 and 8 mL/min), and solute physicochemical properties (lipophilicity, P, fraction unbound to protein, fu; fraction ionized, fi; and molecular weight MW) were varied to better understand the underlying determinants of solute distribution. An apparent low availability was found for a number of the solutes as a consequence of tissue sequestration. This low availability precludes the estimation of an apparent volume of distribution (V) for these solutes. The V of solute and tissue (skin, fat, and muscle) blood flow increased with perfusion flow rate (p < 0.01). The unbound distribution volume (Vu) of basic solutes was significantly linear with respect to P. Multiple linear regression analysis showed that the distribution volume of solute in tissue was significantly related to fu (p < 0.01), but not improved by including relationships to P, MW, and fi. Data obtained with this IR technique yield results consistent with in vivo studies in terms of the importance of fu as a determinant of V. This work has shown that the estimations of solute V by the IR technique in a single-pass preparation are unreliable for solutes with a low availability due to apparent solute sequestration into tissue. The parameter V may also be affected by changes in the perfused limb physiology associated with the perfusion conditions used. The Vs for lidocaine and diazepam vary with fu in accordance with deductions based on the results of steady-state studies. PMID- 7500270 TI - Inclusion complexation of heptakis(2,6-di-O-ethyl)-beta-cyclodextrin with tiaprofenic acid: pharmacokinetic consequences of a pH-dependent release and stereoselective dissolution. AB - beta-Cyclodextrin was ethylated at the 2- and 6-hydroxyl positions. Diethyl sulfate was employed as an alkylating reagent. NMR spectra data indicate that heptakis(2,6-di-O-ethyl)-beta-cyclodextrin (DCD) is the principal component of the product obtained. In addition, the FAB mass spectra obtained in nitrobenzyl alcohol and glycerin matrices gave pseudo-molecular ions with m/z ratios of 1630.75 and 1711.90 corresponding to C70H126O35[2Na.NaCl] and C70H126O35[2C3H8O3], respectively. The dissolution of tiaprofenic acid (TA) enantiomers, from TA powder (10 mg) and inclusion complex and/or coprecipitate (IC) (TA:DCD 1:1 molar ratio, equivalent to 10 mg of free TA), were examined using the dispersion method at pH values of 1.5, 3.0, and 7.4. Complex formation with the hydrophobic DCD resulted in a significant reduction in the release rate of both R- and S-TA, as compared to that observed with the powder. At pH 1.5, tiaprofenic acid enantiomers were not released from IC, compared to 20.52 +/- 1.47% of R-TA and 20.47 +/- 1.64% of S-TA dissolved from the powder. The greatest stereoselectivity in release profiles was found at pH 3.0 from IC [S:R 24 h cumulative percent release (sigma R24) ratio of 0.88 +/- 0.04]. Elevation of the pH to 7.4, which resulted in a faster dissolution and greater sigma R24 of enantiomers from both powder and IC, was accompanied by a parallel reduction in the stereoselectivity. Following single 20 mg/kg oral doses of racemic TA as both powder or IC to Sprague-Dawley rats, significant stereoselectivity was observed in the plasma concentration profiles of the enantiomers (S:R AUC(0-infinity) = 1.5). Despite significant reduction in the rate and extent of absorption, there was not a significant difference in the observed in vivo stereoselectivity between the two formulations. Therefore the in vivo importance of the observed stereoselectivity in release at pH 3.0 is ruled out. Nevertheless, consideration must be given to the possibility of stereoselective release when chiral excipients are used in the formulation of racemic drugs. PMID- 7500274 TI - USP dissolution apparatus 3 (reciprocating cylinder): instrument parameter effects on drug release from sustained release formulations. AB - The United States Pharmacopoeia dissolution apparatus 3 (reciprocating cylinder) was evaluated with respect to effects of changes in instrument parameters on drug release rate from six hydrophilic matrix formulations and one coated-bead formulation. Reciprocation rate had the largest effect on time to 50% release for all matrix formulations. Top mesh size had an effect on release rate for those formulations having an erosion component to the drug release mechanism. Bottom mesh size had no effect on release rate. For the coated-bead formulation, no parameter had an effect on release rate. In an assessment of the hydrodynamics of the reciprocating cylinders, conditions equivalent to the 50 rpm paddle and 100 rpm basket were determined to be at the extreme low end of the available reciprocation rate range. PMID- 7500275 TI - Preliminary safety evaluation of parenterally administered sulfoalkyl ether beta cyclodextrin derivatives. AB - Parenteral administration of beta-cyclodextrin (beta-CD) results in renal and/or local toxicity dependent on the mode of administration. In an attempt to alleviate these properties, a series of anionically charged sulfoalkyl ether cyclodextrin (SAE-beta-CD) derivatives have been developed. The parenteral safety of these derivatives was determined by survival of male mice after intraperitoneal (ip) injection, kidney histopathology, plasma urea nitrogen levels of mice determined 24 h after injection, relative in vitro hemolytic potential and activated partial thromboplastin times (APTT). In addition, the 24 h renal excretion behavior of the derivatives was measured. Where appropriate, the results obtained with these cyclodextrin derivatives were compared with results obtained for beta-CD and (hydroxypropyl)-beta-cyclodextrin (HP-beta-CD). The SAE-beta-CD derivatives did not produce mortality in mice following ip injection at doses exceeding 5.45 mmol/kg. No significant histological lesions were observed in the kidney tissue of mice receiving the cyclodextrin derivatives. The SAE-beta-CD derivatives were excreted faster and to a greater extent than beta-CD and at rates comparable to HP-beta-CD. The hemolytic potential of these derivatives was less than that of beta-CD and comparable to or better than that of HP-beta-CD. The SAE-beta-CD derivatives did not increase APTT clotting times indicating that these derivatives have no significant anticoagulant activity. The toxicological profile of these derivatives suggests that these molecules may have application as biologically safe beta-CD derivatives. PMID- 7500276 TI - The disposition, tissue distribution, and cellular transport of N [[(trimethylamino)boryl]carbonyl]-L-phenylalanine methyl ester in CF1 mice. AB - The disposition and tissue distribution of 14C-labeled N [[(trimethylamino)boryl]carbonyl]-L-phenylalanine methyl esther (1) determined in CF1 mice following i.v., p.o., and i.p. administration. Compound 1 was found to undergo rapid and extensive metabolism, and the majority of the radioactivity was found in the skin and carcass regardless of the route of administration. Approximately 55% of the radioactivity was recovered in urine and feces after 78 h; however, excretion via these routes was not complete. Degradation of compound 1 occurred at the amide bond at low pH (0.8) but at the ester bond a physiological pH. This difference in degradation was reflected in elevation of the blood radioactivity levels after p.o. administration compared to i.p. and i.v. administration. PMID- 7500278 TI - Enhanced cellular uptake and in vivo biodistribution of a monoclonal antibody following cationization. AB - Owing to the poor transport of monoclonal antibodies across either capillary or cell membrane barriers, drug delivery strategies are needed to target monoclonal antibodies to intracellular sites where proteins function. One antibody drug delivery strategy is cationization, wherein the isoelectric point (pl) is raised by conversion of surface carboxyl groups to extended primary amino groups. The present studies describe the cationization of a murine monoclonal antibody (D146) prepared against a synthetic peptide encoding the Asp13 point mutation of the ras protooncogenic p21 protein. The pl of the D146 monoclonal antibody was raised from 8.9 to > 9.5. The uptake in vitro of the 125I-labeled native D146 antibody by MDA-MB231 human carcinoma cells was negligible, whereas there was a marked increase in the endocytosis of the antibody following cationization. In vivo pharmacokinetic analysis was performed in male BALB/c mice. The in vivo organ uptake of the cationized monoclonal antibody was increased relative to the native antibody; there was a 6-fold increase in the systemic volume of distribution, a 58-fold increase in the systemic clearance of the cationized antibody from the plasma compartment, and a 9-fold reduction in the mean residence time of the cationized antibody as compared to the native D146 antibody. In conclusion, these studies show that cationization of an oncogene-specific monoclonal antibody results in markedly increased endocytosis of the antibody by cancer cells in vitro and in increased systemic clearance and organ uptake in vivo. PMID- 7500277 TI - Enantiomeric resolution with a new chiral stationary phase of 7-chloro-3-methyl 3,4-dihydro-2H-1,2,4-benzothiadiazine S,S-dioxide, a cognition-enhancing benzothiadiazine derivative. AB - The direct analytical and semipreparative high-performance liquid chromatographic (HPLC) resolution of the enantiomers of IDRA 21 [1,7-chloro-3-methyl-3,4-dihydro 2H-1,2,4-benzothiadiazine S,S-dioxide] is reported. (+/-)-IDRA 21 administered orally to rats subjected to a water maze cognition test elicited a performance enhancing effect. Between the two enantiomers, (+)-IDRA 21 was identified as being pharmacologically active in the water maze performance test, whereas (-) IDRA 21 was completely devoid of activity when given in doses comparable to those of the dextrorotatory (+)-enantiomer. The design and preparation of a new chiral stationary phase (CSP) employed for the liquid chromatographic resolution of the enantiomers of racemic IDRA 21 is presented. This brush-type CSP, which has not been described before, is a "mixed" (pi-donor pi-acceptor) type and is derived from (R)-N-(3,5-dinitrobenzoyl) allylglycine 2,6-dimethylanilide. It is easily prepared and possesses a relatively broad scope of applicability, as determined by its ability to resolve the enantiomers of both pi-acidic and pi-basic compounds. PMID- 7500279 TI - Location of binding sites on immobilized human serum albumin for some nonsteroidal anti-inflammatory drugs. AB - Anti-inflammatory drugs are widely used therapeutic agents and are very often administered with various other drugs. Because they are highly bound to human serum albumin (HSA), interferences between nonsteroidal anti-inflammatory drugs (NSAIDs) and coadministered drugs may arise from their interactions at a binding site on HSA. Although the percentage of binding to HSA is generally accurately determined, the binding sites to which a particular therapeutic agent binds are often unknown. In order to clarify where different classes of NSAIDs bind on the HSA molecule, competition studies were carried out on a HSA-based HPLC column using site I and site II markers as displacing agents. Results show that all the NSAIDs included in the study were affected by site I and site II markers and that a number of drugs had (an) extra binding site(s) not affected by any of the competitors used in the study. Competition data also suggest that binding of NSAIDs at the benzodiazepine site could in fact occur at two separate subsites, as previously observed for benzodiazepines. PMID- 7500280 TI - Physicochemical properties of A-75998, an antagonist of luteinizing hormone releasing hormone. AB - The physicochemical properties of A-75998, a synthetic antagonist of luteinizing hormone releasing hormone with potential for treatment of hormone-sensitive cancers and endometriosis, are described. An accelerated solution stability study indicated that the compound is relatively stable and showed a U-shaped pH-rate profile, with maximum stability between pH 4.5 and 6.5. The acid dissociation behavior of A-75998 was examined by UV-visible spectrophotometry at 25 degrees C in a series of buffers ranging from pH 1 to 13. The data were fit to a model in which the dissociations of all four ionizable groups contributed to changes in the absorbance. The estimated macroscopic acid dissociation constants were p beta 1 = 3.230 +/- 0.022, p beta 2 = 4.885 +/- 0.030, p beta 3 = 9.871 +/- 0.022, and p beta 4 = 11.026 +/- 0.157. The corresponding microscopic dissociation constants were pk1 = 3.24 (nicotinyl), pk2 = 4.88 (pyridyl), pk5 = 9.91 (tyrosyl), and pk6 = 10.99 (isopropyllysyl). The apparent n-octanol/water partition coefficients were measured from pH 2 to 13, and the profile was consistent with the expected acid-dissociation behavior. While appearing fairly water-soluble at pH < 5, dynamic light scattering of A-75998 in pH 4.5 buffer indicated the formation of aggregates of nonuniform size distribution. A-75998 exhibited reverse or thermal gelation; sodium chloride exacerbates this gel formation and self-association. Surface activity was pH-dependent, but no evidence was found for micelle formation. Based on the results, development of a parenteral formulation of A 75998 appears feasible, provided that aggregation can be minimized. PMID- 7500282 TI - Inferring thermodynamic stability relationship of polymorphs from melting data. AB - This study investigates the possibility of inferring the thermodynamic stability relationship of polymorphs from their melting data. Thermodynamic formulas are derived for calculating the Gibbs free energy difference (delta G) between two polymorphs and its temperature slope from mainly the temperatures and heats of melting. This information is then used to estimate delta G, thus relative stability, at other temperatures by extrapolation. Both linear and nonlinear extrapolations are considered. Extrapolating delta G to zero gives an estimation of the transition (or virtual transition) temperature, from which the presence of monotropy or enantiotropy is inferred. This procedure is analogous to the use of solubility data measured near the ambient temperature to estimate a transition point at higher temperature. For several systems examined, the two methods are in good agreement. The qualitative rule introduced this way for inferring the presence of monotropy or enantiotropy is approximately the same as The Heat of Fusion Rule introduced previously on a statistical mechanical basis. This method is applied to 96 pairs of polymorphs from the literature. In most cases, the result agrees with the previous determination. The deviation of the calculated transition temperatures from their previous values (n = 18) is 2% on average and 7% at maximum. PMID- 7500281 TI - Oral absorption of anti-AIDS nucleoside analogues. 1. Intestinal transport of didanosine in rat and rabbit preparations. AB - The intestinal transport of didanosine (ddl), a nucleoside analog used in the treatment of human immunodeficiency virus (HIV) infection, was characterized using in situ and in vitro techniques. The zero-trans uptake of ddl in rat intestinal brush border membrane vesicles (BBMV) was linear over the range of 1 microM to 50 mM, ruling out a significant carrier-mediated absorption component. The lack of carrier-mediated transport was confirmed in a second species (rabbit). In order to quantitate the convective (Pconv) and diffusive (Pdiff) components of ddl intestinal permeability, the steady state wall permeability (P*w) was determined using an established perfusion technique in rats. Even though baseline P*w (pH 6.5, 290 mosm/kg, no modulator) and fluid absorption results were similar to those of furosemide, the ratios (ddl:furosemide) of Pdiff and phi, the sieving coefficient, were 0.31:1 and 1.70:1, respectively, demonstrating that ddl's Pdiff is low and Pconv is high relative to furosemide's, suggesting significant paracellular absorption of ddl. The apparent diffusive absorptive clearances (P'app) of ddl and furosemide were determined in BBMV, which lack functional tight junctions, and the ratios (ddl:furosemide) of P'app in rat and rabbit were 0.23:1 and 0.24:1, respectively. The BBMV results demonstrate that the majority of ddl intestinal transport does not occur by passive membrane diffusion, confirming the single pass intestinal perfusion (SPIP) findings. The results of these studies suggest that ddl is transported by nonfacilitated membrane and paracellular diffusion with paracellular transport being responsible for the majority of ddl absorption from the intestine. PMID- 7500283 TI - The effect of temperature upon the permeation of polar and ionic solutes through human epidermal membrane. AB - The temperature dependence of in vitro permeation through human epidermal membrane (HEM) was determined for urea, mannitol, tetraethylammonium ion (TEA), and corticosterone. The effect of temperature upon HEM electrical resistance was also measured. The majority of the experiments involved measuring the permeability coefficients of a specific permeant at 27 degrees C and 39 degrees C for a given HEM sample, the electrical resistance was also measured at each temperature. Similar experiments were also conducted with a model synthetic porous membrane. The effect of temperature was quantitated as the ratio of the permeability at 39 degrees C to the permeability at 27 degrees C for each permeant. These ratios observed for HEM with urea, mannitol, and TEA as the permeants were 1.66 +/- 0.05, 1.76 +/- 0.14, and 1.71 +/- 0.11, respectively. The change in temperature was shown to have a similar effect upon the electrical conductance of the HEM samples. The observed ratio for corticosterone permeation was 4.5 +/- 0.4. The experimental ratios observed for the three polar/ionic permeants were shown to approach those obtained from the model porous membrane and differed greatly from the ratio observed for the more lipophilic corticosterone, indicating differences in the effective transport mechanism/pathway for these classes of permeants. The permeability of urea was also observed to be inversely proportional to the electrical resistance of the HEM samples; this relationship was shown to be independent of temperature over the temperature range studied. The temperature dependence data and the observed relationship between urea permeability and electrical resistance strongly support the existence of a porous permeation pathway through the HEM as an operative diffusional route for polar-ionic permeants. PMID- 7500284 TI - Inhibition of indomethacin crystallization in poly(vinylpyrrolidone) coprecipitates. AB - Differential scanning calorimetry and powder X-ray diffraction studies have been carried out with amorphous coprecipitates of indomethacin and poly(vinylpyrrolidone), PVP, to measure the glass transition temperature, Tg, as a function of mixture composition and the nonisothermal and isothermal crystallization of the indomethacin. Values of Tg as a function of mixture composition followed the ideal Gordon-Taylor equation up to about 50% w/w PVP. Inhibition of crystallization occurred at levels as low as 5% PVP and very significant inhibition was observed at and above 20% PVP. Inhibition of crystallization of indomethacin in the absence of PVP required a storage temperature 40-50 degrees C below Tg, whereas comparable inhibition with PVP was observed at storage temperatures 5 degrees C above Tg. This suggests that the inhibition of indomethacin crystallization by PVP may involve mechanisms other than just the general antiplasticizing effect (raising Tg) by PVP. PMID- 7500285 TI - Permeability of pure enantiomers of ketorolac through human cadaver skin. AB - The permeability of pure enantiomers of ketorolac acid, a potent non-narcotic analgesic, through human cadaver skin was evaluated. The melting temperature of each enantiomer was 20 degrees C higher than that of the racemic compound. As expected, the solubility of the racemic compound in water and isopropyl alcohol/water/isopropyl myristate (IPA/water/IPM, 50:50:1.5) was roughly 2 times higher than that of the enantiomers. The permeability of the enantiomers through poly(ethylenevinyl acetate) (EVA) synthetic membrane and human cadaver skin was determined with a side-by-side diffusion cell. The skin flux of the racemic compound was about 1.5 times higher than those of the enantiomers. On the other hand, no significant differences in the intrinsic permeability coefficient of the racemic compound and the enantiomers in the EVA membrane and human cadaver skin was observed. An excellent agreement between the predicted and experimental flux ratio of the racemic compound and enantiomer in the EVA membrane and cadaver skin was observed. The IPA/water/IPM (50:50:1.5) provided the highest in vitro skin flux of the S enantiomer among the three vehicle formulations studied. The skin flux of the active pure S enantiomer was ca. 34% higher than that of the impure S enantiomer in the racemic mixture. Furthermore, about 14% intersubject variability in the in vitro skin flux of the S enantiomer was observed. The required skin flux of the S enantiomer as calculated from the pharmacokinetic parameters was about 32 micrograms/cm2/h from a 25 cm2 transdermal patch, which was readily achievable from the IPA/water/IPM (50:50:1.5) ternary vehicle system. PMID- 7500286 TI - Drug diffusion front movement is important in drug release control from swellable matrix tablets. AB - Swellable controlled release devices of buflomedil pyridoxalphosphate in hydroxypropyl methylcellulose were prepared, and their swelling and release behavior was investigated. The drug release as a function of time was investigated for various system parameters. Three distinct fronts were observed during the swelling and release processes, i.e., a swelling, a drug diffusion, and an erosion front. The drug diffusion front could be readily determined due to the drug's yellow color. The relative positions of the fronts and the drug release rate were studied as functions of the initial porosity and the molecular weight of the polymer carrier. It was shown that the drug diffusion front best describes the overall release behavior of the system. The fractional drug release was a strong function of the dissolved drug gel layer thickness, which separates the diffusion front from the erosion front. The effect of drug solubility was also investigated by altering the pH and the ionic strength of the dissolution medium. It was shown that as drug solubility increased, the undissolved drug gel layer thickness decreased, again showing the importance of the movement of the diffusion front in controlling the overall release. PMID- 7500287 TI - Acid hydrolysis of diazepam. Kinetic study of the reactions of 2-(N-methylamino) 5-chlorobenzophenone, with HCl in MeOH-H2O. AB - In the acid hydrolysis of diazepam (1), several unusual products, apart from 2-(N methylamino)-5-chlorobenzophenone (2) and glycine, were isolated. On the assumption that some of those products could arise from further degradation of 2, the reaction of this compound with 0.5-2 M HCl was studied, in 1:1 MeOH-H2O, at 60 and 80 degrees C. Several unexpected products were isolated from the reaction of 2 with HCl, namely, 2-amino-5-chlorobenzophenone (3), 2-(N,N-dimethylamino)-5 chlorobenzophenone (4), 2-(N-methylamino)-3,5-dichlorobenzophenone (5), 2-amino 3,5-dichlorobenzophenone (6), 2,4-dichloro-10-methyl-9,10- acridinone (7), and 2,4-dichloro-9,10-acridinone (8). The methyl transfers, the chlorination, and the cyclization reactions that give rise to products 3-8 are unexpected under the present reaction conditions. The rate of reaction of 2, as well as the rate of formation of compounds 3-6, was measured at several HCl concentrations. PMID- 7500288 TI - Expression of p53 protein in oral squamous cell carcinomas and adjacent non tumorous mucosa of the floor of the mouth: an archival immunohistochemical study using wet autoclave pretreatment for antigen retrieval. AB - Routinely formalin-fixed and paraffin-embedded material of 22 squamous cell carcinomas of the floor of the mouth (T2NoMo, Ro), together with adjacent dysplastic or normal mucosa, were immunohistochemically investigated using a panel of four anti-p53 antibodies (CM1, PAb1801, DO7, PAb240) subsequent to wet autoclave pretreatment for antigen retrieval. p53 immunoreactivity was detected in 9/22 (40%) carcinomas with PAb1801 and DO7 antibodies, and in 8/22 cases using CM1 and PAb 240. p53-positive tumour cells accumulated predominantly at the invasive front of the carcinomas. A focal or scattered p53 immunoreactivity was observed in the adjacent normal and/or dysplastic mucosa in 17/22 (77%) cases using both CM1 and PAb1801 antibodies, in 10/22 with DO7, and in 8/22 with PAb240. This study has demonstrated examples of different p53 immunophenotypes in the non-tumorous and neoplastic oral mucosa of the same patient without significant correlation to the clinicopathological parameters studied. PMID- 7500289 TI - Integrin receptors and their relationship to cellular proliferation and differentiation of oral squamous cell carcinoma. A quantitative immunohistochemical study. AB - The expression of extracellular matrix (ECM) proteins (fibronectin, laminin, collagen IV) and ECM receptors of integrin type (alpha 2 beta 1, collagen receptor; alpha 6 chain of the fibronectin receptor; alpha 6 chain of the laminin receptor) were examined in normal oral squamous epithelium and in invasive areas of squamous cell carcinomas with various differentiation and proliferation activities (Ki-67 antigen labelling), evaluating the presence, quantity (using an image analysis system) and distribution of the integrin subunits. In the mucosa, there was uniform immunostaining for alpha 2 beta 1 and alpha 6 concentrated at the cell membrane in the basal/supra basal cell zone, whereas, alpha 5 showed a discontinuous staining of the basal cell-basement membrane interface. alpha 2 and alpha 6 could be visualized in all carcinomas. alpha 5 showed low expression preferentially in less differentiated carcinomas. In contrast to normal mucosa, there was an increase in alpha 6 staining in well-differentiated carcinomas. Dedifferentiation of oral carcinomas was accompanied by an increase in cellular proliferation and with a decrease in alpha 2 beta 1 and alpha 6 staining. This reduction of alpha 6 staining was shown to be statistically significant, suggesting that this integrin may be a valuable grading parameter for oral squamous cell carcinoma. PMID- 7500290 TI - Growth of oral and skin fibroblasts from patients with oral submucous fibrosis. AB - The purpose of the investigation was to evaluate and compare the proliferation (growth) of mouth fibroblasts and skin fibroblasts from patients with oral submucous fibrosis (OSF). Material comprised fibroblasts from fibrous bands situated in the buccal mucosa and from the inner aspect of the forearm of 8 patients with classic features of OSF as well as fibroblasts from 6 buccal mucosa and 8 skin biopsy specimens from healthy non-areca nut chewing individuals. Cells were cultured for 8 days according to standard techniques. Their growth was monitored daily, under optimal conditions as well as exposure to concentrations of arecoline. The data were analyzed using regression analysis, analysis of variance and the Kruskal-Wallis test. We found no statistically significant differences between the proliferation patterns of oral and skin fibroblasts from patients or between those from patients and controls. The reaction of the cells exposed to concentrations of arecoline was similar; at low concentrations (0.1-10 micrograms/ml) normal growth was maintained, while 100 micrograms/ml inhibited growth. It is concluded that fibroblasts from mouths affected by OSF have proliferation patterns which fall within normal parameters, that the excessive collagen formation in established OSF is not due to increased fibroblast proliferation and that arecoline does not stimulate fibroblast proliferation. PMID- 7500292 TI - Oral candidal flora in healthy infants. AB - Asymptomatic oral candidal carriage has been extensively studied in adults, but only rarely in infants. The present investigation aimed to determine the asymptomatic oral carriage of candidal species in healthy infants and its relationship to age, sex, feeding pattern and use of a pacifier. The swab technique was used for oral candidal isolation. Candidal species were isolated from 48% of the infants without a significant relationship to age, gender or between breast-fed or bottle-fed infants. Similarly, a history of maternal vaginal candidosis, or presence of nappy rash, had no significant relationship to oral candidal carriage. Infants who routinely sucked a pacifier had a significantly higher rate of oral candidal carriage, suggesting a reservoir of infection. PMID- 7500291 TI - High prevalence of hepatitis C virus antibody and RNA in patients with oral cancer. AB - We have investigated the correlation between the prevalence of hepatitis C virus (HCV), which is detectable in saliva, and oral cancer and other digestive tract cancers in the Northern Kyushu region of Japan. Anti-HCV antibodies were detected in sera from 24 of the 100 patients with oral cancer (24%, p < 0.05 vs the control group, p < 0.01 vs the stomach cancer group), in 11 of 104 patients with non-malignant diseases receiving dental treatment (the control group, 10.6%), and in 12 of 113 patients with stomach cancer (10.6%). HCV-RNA was detected in sera from 17 of 100 oral cancer patients (17%, p < 0.05 vs the control group) and 4 of 104 patients of the control group (3.9%). These results indicate a high prevalence of HCV infection in oral cancer patients, which warrants a systematic study of etiological associations between oral cancer and HCV. PMID- 7500293 TI - Interleukin-1 (IL-1) bioactivity and inhibition of IL-1 bioactivity in supernatant fluid from cultured microphthalmic (mi) mice teeth of different ages. AB - A functioning dental follicle and bone resorption are necessary for tooth eruption. In the microphthalmic (mi) mouse, bone resorption is defective and teeth fail to erupt. Local bone metabolism involves the production of cytokines such as interleukin-1 alpha (IL-1 alpha) and transforming growth factor-beta (TGF beta). The production of these cytokines by dental tissues during eruption may be fundamental to tooth movement. Molars from mi mice were cultured and supernatants tested for the presence of these cytokines using bio-dot blotting and ELISA. A thymocyte bioassay was used to test supernatants for IL-1 bioactivity and IL-1 inhibition bioactivity. IL-1 alpha and TGF-beta were detected in all supernatants. Supernatants demonstrated no IL-1 bioactivity but inhibited IL-1 bioactivity which varied with concentration of supernatant, age and animal. This study demonstrated that cultured developing teeth secrete IL-1 alpha and TGF-beta however, concentrations varied in normal and pathological states. While IL-1 alpha was present in the supernatants, all demonstrated a variable ability to inhibit IL-1 bioactivity. This ability may influence local bone metabolism and hence tooth eruption. PMID- 7500294 TI - Linear IgA disease (LAD): immunoglobulin deposition in oral and colonic lesions. AB - Two cases of adult linear IgA disease (LAD) with oral and colonic involvement are presented. Oral findings, for both patients, were of widespread painful ulceration present for two to three months and diagnosis was confirmed by biopsy and direct immunofluorescence. Colonic symptoms preceded oral symptoms in both cases and a number of investigations were undertaken, including routine histological examination, which suggested either lymphocytic colitis or Crohn's disease. Immunofluorescence studies showed linear IgA deposition confined to the distal colon. These cases highlight that large bowel involvement can be a feature of linear IgA disease and for the first time demonstrate IgA deposition at that site. PMID- 7500295 TI - Spindle-cell hemangioendothelioma of the oral cavity. A case report. AB - We describe a vascular tumor classified as SCH by histological criteria that was found in the mandibular-buccal fold of a 12-year-old girl. Microscopically, the lesion consisted of thin-walled cavernous spaces containing thrombi and phleboliths, and cellular areas composed of spindle-shaped, epithelioid and vacuolated cells. Immunohistochemically, the endothelial vascular lining was highly reactive with HAM56 antibody, while variable reactivity was observed for factor VIII-associated antigen. All cell types were positive for vimentin and anti-PCNA stained less than 3% of the tumor cells. This is the first report of SCH in the oral cavity. PMID- 7500296 TI - Proteus syndrome: association with gingival hyperplasia. AB - A 9-year old Black boy with gigantism of the hands and feet, and recurrent gingival hyperplasia, diagnosed as Proteus syndrome is presented. The oral manifestations of this syndrome are described. To the best of our knowledge, this is the first reported case of gingival hyperplasia associated with Proteus syndrome. PMID- 7500297 TI - Liberal or conservative? Gender, identity, and perception of historical religious positions. AB - Conservatism is often assumed to imply a continuity of values and positions between past and present. Previous research has demonstrated, however, that there are discontinuities between historical and contemporary political conservatism and that modern political conservatives endorse programs and legislation once regarded as liberal. Testing whether the same pattern may hold for religious conservatism was the purpose of this research. Perceptions of conservative positions in Christian history were assessed among young adults (N = 221). Results showed that individuals who identified themselves as conservatives were less likely to recognize past conservative positions on religious issues than individuals who identified themselves as liberals. These findings were not accounted for by lack of knowledge about religious history on the part of conservatives. Furthermore, women were more likely than men to accurately identify conservative and liberal positions of the past. The findings suggest that religious conservatism, like political conservatism, does not necessarily involve a continuity of positions between past and present. PMID- 7500298 TI - Relationships among job involvement, job satisfaction, and organizational commitment for nurses. AB - The relationships among a cluster of attitudes toward work and job were investigated using a sample of 171 nurses. The hypothesis was that involvement in work and job, commitment to the employing organization, and satisfaction with the job (overall, and with specific facets of the job) would be significantly correlated. The results showed that involvement was not related to overall satisfaction but only to two specific facets, satisfaction with work and promotion opportunities. In contrast, the degree of relationship between overall and various facets of satisfaction and commitment and between involvement and commitment was moderately high. PMID- 7500299 TI - Effects of adult attachment style on the perception and search for social support. AB - Effects of adult attachment style on the perception of and search for social support were examined. One hundred and fifty undergraduate students completed self-report scales tapping their attachment styles, the extent to which they perceive the availability of emotional and instrumental support from significant figures (father, mother, same-sex friend, opposite-sex friend, romantic partner), and the extent to which they look for instrumental and emotional assistance in times of need. Findings showed that secure persons perceived higher levels of emotional and instrumental support from the assessed figures and reported seeking more emotional and instrumental support than avoidant and ambivalent persons did. Results are discussed in the context of an attachment perspective of social support. PMID- 7500300 TI - The perception of risk over time as a factor influencing attitudes toward children with physical disabilities. AB - The impact of perception of risk (a perception of possible psychological harm) on attitudes toward friendship with and offering help to children with physical disabilities was examined, and the stability of the perception in specific contexts, over time, was investigated. Participants were 510 children without physical disabilities between the ages of 8 and 19 years who were assigned to 1 of 12 experimental conditions in a basic 2 x 2 x 3 (Involvement x Interview x Grade) design. Results indicated significantly more favorable attitudes on helping than on friendship. Over time, attitudes toward friendship became more positive, whereas attitudes toward helping became somewhat more negative, but these effects interacted with the age of the participant and the degree of perceived risk. PMID- 7500301 TI - Delayed reporting of the rape victim. AB - 1. Delayed reporting of rape may be due to impaired cognitive processing, altered states of consciousness, or cognitive dissonance. 2. DNA may speak for the victim who does not remember a rape due to being unconscious. 3. Women with psychosis may incorporate a rape into their delusional system. PMID- 7500302 TI - Infant abductors. AB - 1. Admitted kidnappers identified four phases involved in stealing an infant as: setting the stage for a baby, planning the abduction, the act of abduction, and post-abduction discovery. 2. Abductors describe personal pressure and interpersonal pressures as motivations in stealing an infant. 3. The act of abducting an infant ranges from the use of no force to lethal force. PMID- 7500303 TI - Investigating stalking crimes. AB - 1. Three types of stalkers that are identified in this crime classification are the non-domestic stalker who may know the target through social contact or from a random meeting in a public place; the domestic stalker who may be known to the target and had a close personal relationship with the target; and the erotomania stalker whose target is typically a public figure. 2. Stalking is conceptualized on a continuum from nondelusional to delusional behavior. 3. Classification of a crime is the first step in the investigative process. PMID- 7500304 TI - Transgendered ... and taken to jail. AB - 1. Gender identity disorders have the essential feature of incongruence between assigned sex, that is, the sex that is recorded on the birth certificate, and gender identity. 2. The majority of transsexuals are males who consider themselves to have feminine gender identity and regard their genitalia and masculine features with repugnance. 3. Intersexuality occurs when gender identity arises when the child is born with ambiguous genitalia. PMID- 7500306 TI - Nursing interventions in Munchausen syndrome by proxy. AB - 1. Munchausen syndrome by proxy is a rare but serious and potentially lethal form of child abuse in which a primary care giver induces or reports factitious symptoms in a child. 2. The child suffers from the caregiver's actions and health care providers unknowingly become accomplices when they provide unnecessary testing and therapies. 3. The child's physical and psychological welfare can be protected through the efforts of the forensic nurse specialist coordinating a multidisciplinary health care team's work to detect the syndrome and intervene early on. PMID- 7500307 TI - Forensic nursing and death investigation: will the vision be co-opted? AB - 1. When entering allied fields, nurses must hold fast to their unique skills as nurses. 2. Nurses investigating deaths must be cooperative with other disciplines, but must not be co-opted by them. 3. Caring and empathy are integral and significant parts of the nursing profession that fulfills an important human need in death investigation. PMID- 7500305 TI - Shaken baby syndrome. AB - 1. Child abuse, in the form of Shaken Baby syndrome, has reached a critical level. 2. Education on a national level is crucial for change to help save the victims of Shaken Baby syndrome. The national statistics for child abuse reinforce that the United States must deal with the issue and focus on education. 3. Health care providers are in a strategic position to offer primary and secondary prevention interventions, to identify families at risk, and to recognize victims of Shaken Baby syndrome. PMID- 7500308 TI - Forensic nursing: what's new? PMID- 7500309 TI - Memory presentations of childhood sexual abuse. AB - Questions are continually raised about the accuracy and validity of very young children's memories of traumatic events. Out of 19 children, where the median age was 2 1/2 at time of disclosure, 11 had full verbal memory, five had fragmented verbal memory traces, and three had no memory 5 to 10 years following day care sexual abuse. Data from this clinical study suggest the nature of children's memory is four-dimensional: somatic, behavioral, verbal, and visual. Efforts need to continue to document the nonverbal components for assessment and treatment purposes. PMID- 7500310 TI - Immersion fatalities: hazardous responses and dangerous discrepancies. AB - In this paper the following question is addressed: 'Why, given the plethora of standards, specifications and guidelines for immersion protective equipment, are lives still being lost at sea and fatal accident enquiries questioning the quality of such equipment?' In attempting to answer this question, consideration is given to the extent to which both the possible prevailing environmental conditions, and the physiological responses they evoke, are recognised in the design, selection and evaluation of immersion protective equipment. The hazardous responses associated with immersion in cold water are briefly reviewed and the value and relevance of some of the existing tests of immersion protective clothing are considered. It is concluded that: i. when standards, policies and tests for the selection and use of immersion protective clothing are being formulated, consideration should be given to all of the hazardous responses associated with immersion; ii. it should be recognised that the performance of immersion protective equipment during an accident may be significantly inferior to that predicted by routine testing for certification. PMID- 7500311 TI - Drowning and near drowning. PMID- 7500312 TI - Repeated pseudo-obstruction, mental subnormality--and serious snags. AB - Chronic intestinal pseudo-obstruction has certain identifiable causes but many cases remain idiopathic. The majority of patients are managed successfully by conservative means with the avoidance of surgery. A case is presented where diagnosis was made difficult by autism and which was complicated by perforation of a gastric ulcer. Management of pseudo-obstruction is discussed and emphasis placed on an awareness of alternative life-threatening pathologies at re presentation. PMID- 7500313 TI - Aspects of neurological decompression illness: a view from Bethesda. AB - A minority of divers with neurological decompression illness (DCI) fail to improve with recompression treatment. This is particularly seen in cases where features of severe spinal cord injury develop soon after surfacing. Haemorrhage into the spinal cord is implicated in the pathogenesis of these cases, and evidence is presented that supports the view that the bleeding coincides with shrinkage of autochthonous bubbles. The role of hyperbaric oxygen therapy in the treatment of spinal cord DCI is discussed with reference to possible benefit in ischaemia-reperfusion (I-R) injury. Similarities and differences between the tissue injury of dysbaric and conventional spinal cord injury are outlined. The implications of advances in drug therapy for conventional spinal cord trauma are considered in the context of their potential application to treat neurological DCI. PMID- 7500314 TI - The follow up of acute otitis media in general practice. AB - Acute Otitis Media is a common condition of childhood which potentially has a number of significant sequelae including the development of Otitis Media with Effusion ('Glue Ear'). A general practice based study is described which assesses the implementation of a protocol for the follow up of Acute Otitis Media by the primary care team. The results of this study show that following the introduction of a follow up protocol a significantly improved rate of follow up was achieved with resultant improvements in the identification of 'Glue Ear' and hence more appropriate referral for ENT care. PMID- 7500316 TI - Elective report: summer 1994--American Airlines. PMID- 7500315 TI - Health and safety in Medical Centres; finding the hazards and reducing the risks. PMID- 7500317 TI - The battle for Port-en-Bessin 6-8 June 1944: a medical officer with a Royal Marine Commando. PMID- 7500318 TI - Levels of interferon-gamma and tumor necrosis factor-alpha in sera and cervical mucus of fertile and infertile women: implication in infertility. AB - Concentrations of two immune cytokines, namely interferon-gamma (INF-gamma) and tumor necrosis factor-alpha (TNF-alpha), were determined in the sera and cervical mucus samples of fertile (n = 16), idiopathic infertile (n = 44), and immunoinfertile women (n = 45) to investigate their role, if any, in female infertility. Sera of idiopathic infertile women demonstrated significantly (P < 0.0001) higher levels of INF-gamma compared to those in fertile controls, whether expressed as pg/ml or pg/mg serum protein. Similarly, sera of immunoinfertile women demonstrated significantly (P = 0.0008) higher levels of INF-gamma compared to fertile controls and idiopathic infertile women. Cervical mucus of idiopathic infertile women also demonstrated significantly (P < 0.0001) higher concentrations of INF-gamma compared to those in fertile controls. Cervical mucus of immunoinfertile women demonstrated significantly (P < 0.0001) higher concentrations of INF-gamma compared to those in fertile controls and idiopathic infertile women. INF-gamma levels in serum did not significantly (P > 0.05) correlate (r = 0.12-0.43) with the concentrations in cervical mucus, when all the three groups were compared together. However, when the serum levels were compared with the cervical mucus concentrations by condition, only the idiopathic infertile group showed a significant (P = 0.005) correlation (r = 0.70). Serum levels of TNF-alpha did not differ significantly (P > 0.05) among three groups of women. Cervical mucus concentrations of TNF-alpha, however, varied among groups with levels being significantly (P = 0.04) higher-in idiopathic infertile women compared with fertile controls and in immunoinfertile women significantly (P = 0.0007) higher than in fertile controls as well as idiopathic infertile women. TNF-alpha levels in serum correlated (r = 0.65) significantly (P < 0.001) with the concentrations in cervical mucus when all the three groups were compared together or individually by infertility condition. These findings suggest the involvement of cytokines in infertility, and thus may have potential applications in diagnosis and treatment of female infertility. PMID- 7500319 TI - Soluble tumor necrosis factor receptors and soluble interleukin-6 receptor in fetal and maternal sera, coelomic and amniotic fluids in normal and pre-eclamptic pregnancies. AB - The aim of this study was (a) to measure soluble tumor necrosis factor receptors (sTNF-Rs) and soluble interleukin-6 receptor (sIL-6-R) in coelomic and amniotic fluids, cord and maternal sera in pregnancy and labor, (b) to examine whether the changes in concentrations of biologically active TNF and IL-6 are related to changes in their soluble receptors, and (c) to determine if levels of soluble receptors in pre-eclamptic disorders differ from normal pregnancies at delivery. Materials collected from 206 women during pregnancy and at delivery were analyzed for soluble receptors by enzyme-linked immunosorbent assay (ELISA). All receptors were present in higher concentrations in coelomic than in the corresponding amniotic fluid. Concentrations increased in amniotic fluid from first to second trimester. The level of sIL-6-R then remained unchanged to term, but there was a decrease in the sTNF-Rs which might account for the simultaneous appearance of bioactive TNF. Labor did not affect the concentration of any receptor in amniotic fluid. In maternal serum, sTNF-Rs increased with gestational age and labor in parallel with IL-6. The origin and physiological importance of these soluble receptors are still unknown. In pre-eclamptic disorders p55 sTNF-R was elevated in maternal serum before initiation of labor compared to normal pregnancy. PMID- 7500321 TI - Identification of a 53-kDa antigen in the fibrous sheath of avian spermatozoa. AB - The intracellular fibrous sheath that surrounds the proximal part of the sperm flagellar axoneme in non-passerine birds is structurally different from that of mammals. We raised a monoclonal antibody against the fibrous sheath of cockerel spermatozoa by in vitro immunisation. Indirect immunofluorescence and immunogold labelling showed that the antibody bound specifically to the fibrous sheath. It also labelled the fibrous sheath in quail spermatozoa. In both species the antibody bound an antigen that had a molecular weight of about 53 kDa. In tissue sections from adult cockerel testis the antigen was located in spermatids and spermatozoa with little cross-reactivity with the basal region of the seminiferous epithelium or interstitial tissue. The antibody may prove to be a useful tool in studies of avian spermiogenesis. PMID- 7500320 TI - In vitro production of human anti-sperm antibodies: effect of an oligoclonal antibody (F6) on sperm-egg interaction. AB - A method has been developed to establish lines of transformed lymphocytes able to produce in vitro the same anti-sperm antibodies as those naturally occurring in immuno-infertile individuals. We utilized lymphocytes from a male donor whose serum contained anti-sperm antibodies of the IgG class up to the dilution 1:10,000, as detected by means of immunobead binding. T lymphocytes were separated from B lymphocytes using magnetic beads coated with anti-T antibody. B lymphocytes were then placed at a concentration of 5 x 10(6)/ml in a 96-well plate, stimulated with phytohaemagglutinin (PHA) and transformed with Epstein Barr virus. After a few days, only transformed cells continued growing and these were collected. The supernatant was tested for production of anti-sperm antibodies and those transformed lymphocytes shown to be synthesising antibodies directed against the sperm head and the tail were cloned. We obtained a clone of cells producing antibodies of the IgG1 class directed against the head of the spermatozoon. This oligoclonal antibody (F6) recognized a 58-kDa band from a lysate of sperm membranes and was able to reduce the penetration of zona-free hamster oocytes by capacitated spermatozoa. PMID- 7500322 TI - Cellular immune responses in the pig uterus during pregnancy. AB - In pigs, little is known about the role of the uterine immune system during pregnancy. Immunohistochemical studies were conducted using a panel of monoclonal antibodies to pig leukocytes on uterine tissues taken from gilts after fertile mating and at different stages of pregnancy. Acute inflammation in the endometrium in response to fertile mating which included marked changes in the tissue and immune cell components of the endometrium was observed. Throughout pregnancy the pig uterus contained a substantial population of leukocytes. MHC class II staining was prominent in the endometrium at all stages examined and included macrophages, dendritic and fibroblast-like cells, lymphocytes and the endothelial lining of many uterine blood vessels. The majority of lymphoid cells were CD2+, indicating the prevalence of T cells. In early pregnancy specific changes were seen in the tissue distribution of uterine immune cells. Following placentation distinct cellular changes in the local immune cell environment of the uterus were also observed despite the non-invasive nature of the pig placenta. There appears to be suppression and activation of various immune cell components in the uteri of pregnant pigs. This phenomenon is presumably in response to foetal or trophoblast antigens, suggesting that the local immune system is involved in the uterine response to pregnancy. PMID- 7500323 TI - HLA-DQA1 and HLA-DQB1 haplotypes in aborted fetuses and couples with recurrent spontaneous abortion. AB - HLA haplotypes may be associated with spontaneous abortion through a variety of mechanisms, including maternal hyporesponsiveness to fetal alloantigens, maternal autoimmunity, and HLA-linked t-locus homologues. HLA-DQA1 and HLA-DQB1 haplotypes were determined in 37 couples with a history of recurrent spontaneous abortion (RSAB), 40 of their abortuses, and 20 fertile control couples. The distribution of haplotype frequencies did not differ between control subjects and RSAB wives, RSAB husbands, or abortuses. The frequency of the HLA-DR5-linked haplotype, DQA1*501/DQB1*301, which was considered a marker for immune hyporesponsiveness, did not differ between RSAB wives and control subjects (P = 0.353). The frequency of the autoimmune-associated HLA-DR3-linked haplotype, DQA1*0501/DQB1*0201, did not differ significantly between RSAB wives and control subjects (P = 0.103). The frequency of the DQA1*0201/DQB1*0201 haplotype in RSAB husbands was greater than the 95th percentile confidence limit of the frequency of this haplotype in control subjects. Among seven RSAB husbands who were heterozygous for this haplotype and did not share a DQA1*0201 allele with his wife, the haplotype was transmitted to 6 of 7 abortuses (3.5 expected). Although the small size of this sample precludes drawing conclusions regarding HLA transmission biases in RSAB couples, these data have generated a specific hypothesis regarding the DQA1*0201/DQB1*0201 haplotype that can be investigated in future studies. PMID- 7500325 TI - Family history of hypertension and vasovagal symptoms during blood donation. AB - Vasovagal reactions during blood donation were assessed in 185 individuals with and 298 individuals without a parental history of hypertension. It was predicted that individuals at genetic risk for hypertension would be less likely to faint or suffer pronounced vasovagal reactions, presumably by virtue of altered baroreflex sensitivity. A relationship between a parental history of hypertension and vasovagal responses was observed, albeit modified by blood donation experience and sex. Nurses obtaining blood from inexperienced donors without a parental history of hypertension were significantly more likely to recline the subject's chair than those obtaining blood from inexperienced donors with a parental history of hypertension or experienced donors. Questionnaire data were consistent with these findings. These results may be an interesting reflection of group differences in baroreflex sensitivity and have implications for screening potential donors. PMID- 7500326 TI - Leisure-time physical activity in an African-American population. AB - Despite the many important benefits of leisure-time physical exercise on health, many Americans do not engage in exercise. This is more the case for the African American population. This study assesses the impact of efficacy cognitions on leisure-time physical activity (LTPA) in an African-American sample. Our findings show that LTPA is predicted by the sense of efficacy in this African-American sample. Further analyses address significant factors in the prediction of the sense of efficacy. We find that the general sense of mastery predicts efficacy in this African-American sample. This may be especially significant for the African American population. Also, analysis reveals surprising findings concerning characteristics predictive of efficacy cognitions. The highly educated are least likely to believe in the efficacy of preventive health behavior. PMID- 7500324 TI - Peer and parental influences on adolescent tobacco use. AB - Longitudinal models of the development of adolescent smoking and smokeless tobacco (ST) use were tested for a sample of 643 adolescents, age 14 to 17. The sample was assessed at three time points. Smoking, smokeless tobacco, and other problem behaviors formed a single problem behavior factor. Structural equation modeling indicated that inadequate parental monitoring and association with deviant peers at Time 2 predicted tobacco use at Time 3. When parental and peer smoking at Time 2 were added to the model, each accounted for significant variance in predicting Time 3 smoking, but inadequate parental monitoring and association with deviant peers still accounted for some of the variance in Time 3 smoking. In predicting boys' smokeless tobacco use, monitoring at Time 2 predicted smokeless tobacco use, but only when parental approval of ST use was not included. Fathers approval of ST use at Time 2 predicted ST use at Time 3, while maternal disapproval predicted its use. PMID- 7500327 TI - Socioeconomic status and exercise self-efficacy in late life. AB - Self-efficacy, or assessments about one's ability to carry out particular tasks, has been shown to play a central role in the adoption and maintenance of exercise. The relationship between exercise self-efficacy and socioeconomic status (SES), however, has not been formally developed or tested, and the implications of SES for exercise interventions are not known. We hypothesize pathways through which income, education, and occupation affect self-efficacy and capitalize on the availability of responses from 1944 older HMO enrollees to investigate the direct and indirect associations of SES indicators with exercise self-efficacy. Direct associations of age and education are found. Indirect associations of age, income, education, and occupation operate primarily through previous exercise experience, satisfaction with amount of walking, depression, and outcome expectations. The potentially modifiable nature of exercise outcome expectations (i.e., belief in the benefits of exercise) in combination with its strong association with exercise self-efficacy argue in support of greater consideration of its role in attempts to improve exercise self-efficacy. PMID- 7500328 TI - Cancer awareness and self-examination practices in young men and women. AB - Participants in this study (N = 178) were poorly informed about risk factors, warning signs, and self-examination (SE) practices for two common cancers in young adults, testicular cancer in men and breast cancer in women. Compared to women, men were less likely to know about, see the importance of, or practice SE. We found no relationship between internal locus of control, hypochondriasis, and loneliness, on the one hand, and cancer knowledge and SE, on the other. The best predictors of cancer awareness and SE were fear of developing cancer and self rated confidence that SE was being done correctly. The results are consistent with a health belief model and self-efficacy theory of health behavior. PMID- 7500329 TI - The physical and psychosocial predictors of adolescents' recovery from oral surgery. AB - The purpose of this study was to expand medical models of recovery from surgery in adolescents. Sixty-seven adolescents undergoing third molar surgery and their parents participated. Adolescents' negative affectivity, expectancies about recovery, coping styles, and parents' anticipated encouragement of illness behavior were assessed preoperatively. Extent of surgery was assessed by the oral surgeon. Outcome measures included mouth opening, disability, and pain. Extent of surgery did not predict recovery. After controlling for extent of surgery, the psychosocial parameters accounted for an additional 19% of the variance in mouth opening and 21% of the variance in disability. Adolescents' expectancies about recovery and parents' anticipated "pampering" responses predicted mouth opening. Adolescents' expectancies predicted disability. Expectancies about recovery and parental encouragement of illness behavior add predictive power to models of adolescents' recovery from surgery. Limitations of the study, future research directions, and clinical implications are discussed. PMID- 7500330 TI - First glimpses at structure-function relationships of the nucleocapsid protein of retroviruses. AB - Retroviruses are a family of widespread small animal viruses about 110 nm in diameter, composed of an inner core surrounded by an outer envelope formed of a lipid bilayer of cellular origin in which are anchored viral glycoproteins. The inner core is formed by an outer shell of capsid protein molecules (CA protein) surrounding the dimeric RNA genome in close association with about 2000 molecules of nucleocapsid protein (NC protein) and molecules of reverse transcriptase (RT) and integrase (IN). Conversion of the genomic single-stranded RNA into a double stranded proviral DNA by RT takes place in the nucleocapsid substructure and involves two DNA strand transfers to generate the long terminal repeats (LTR) required for IN-mediated integration of the proviral DNA into the cellular genome and its expression. In this review we have summarized some of the properties and functions of the nucleocapsid protein of the most intensely studied oncoretroviruses (MuLV and ASLV) and lentiviruses (HIV-1). Recent biochemical and genetic data on retroviral NC proteins have shown that this small viral protein endowed with a strong affinity for nucleic acids exhibits nucleic acid annealing and strand transfer activities and is required for the formation of infectious viral particles. These new activities of NC protein are most probably necessary at the early steps of proviral DNA synthesis. The 3-D structures of HIV-1 and MoMuLV NC proteins, deduced from NMR studies, are characterized by a central globular domain with one (MoMuLV) or two (HIV-1) zinc fingers. This should facilitate a rational approach of new anti-HIV therapies based on inhibition of NC protein functions. Due to space limitations and the very abundant literature on retroviruses, references to articles prior to the publication of the second volume of RNA Tumor Viruses in 1985 (Weiss et al., 1985) will be minimal. We also direct the reader to an excellent review which summarizes recent insights into biochemical and structural aspects of the retroviral enzymes PR, RT and IN (Katz & Skalka, 1994). PMID- 7500331 TI - The mitochondrial ClpB homolog Hsp78 cooperates with matrix Hsp70 in maintenance of mitochondrial function. AB - The mitochondrial heat shock protein Hsp78 is a member of the Hsp104/Clp family with unknown function. Saccharomyces cerevisiae deletion mutants of HSP78 show wild-type like growth. We report that deletion of the HSP78 gene in yeast strains with point mutations in the SSC1 gene (encoding matrix Hsp70) led to loss of mitochondrial DNA, indicating that at least one of the heat shock proteins Hsp78 and mt-Hsp70 is needed to maintain a rho+ state of the mitochondrial genome. Mitochondria isolated from these double mutants had a strongly reduced membrane potential, explaining defects in the rate of preprotein import. The lack of Hsp78 led to aggregation of the mutant mt-Hsp70 while other matrix chaperones stayed soluble. We conclude that Hsp78 is required to keep mutant forms of mt-Hsp70 soluble and suggest a cooperation of Hsp78 and mt-Hsp70 in maintenance of essential mitochondrial functions. PMID- 7500332 TI - A novel basis of capsid stabilization by antiviral compounds. AB - Picornaviruses are inactivated by a family of hydrophobic drugs that bind at an internal site in the viral capsid and inhibit viral uncoating. A basis for the capsid stabilization previously unrecognized is revealed by molecular dynamics simulations of the antiviral drug WIN52084s bound to a hydrophobic pocket of solvated human rhinovirus 14. Isothermal compressibilities of the complex and human rhinovirus 14 without the antiviral drug calculated from density fluctuations show that the presence of WIN52084s increases the compressibility of the viral capsid near the antiviral drug. This counterintuitive result is understandable on the basis of the empirical evidence of thermal melting temperatures and protein-folding entropies of globular proteins. Based on this evidence, we propose that a larger compressibility from drug binding confers greater thermal stability to capsid proteins by increasing the conformational entropy of capsids, thereby diminishing the entropy gain with uncoating. We suggest that compressibility is fundamental to the structural integrity of viral capsids and that examination of compressibility and antiviral activity will provide insights into the disassembly process. PMID- 7500333 TI - Regulation of codBA operon expression in Escherichia coli by UTP-dependent reiterative transcription and UTP-sensitive transcriptional start site switching. AB - Reiterative transcription is the repetitive addition of nucleotides to the 3' end of a nascent transcript due to slippage between the transcript and DNA template. Recently, we showed that pyrimidine-mediated regulation of pyrBI operon expression in Escherichia coli occurs, in part, through a mechanism in which induction of UTP-dependent reiterative transcription within the initially transcribed region prevents downstream extension of the nascent transcript to include structural gene sequences. In this study we demonstrate that pyrimidine mediated regulation of codBA operon expression in E. coli also involves UTP dependent reiterative transcription during initiation; however, the mechanism is different from that of the pyrBI operon. The initially transcribed region of the codBA promoter contains the sequence GATTTTTTG (non-template strand). Our results show that transcription is initiated primarily at the first two bases designated G7 and A8 (counting from the -10 region). When transcripts are initiated at position A8, UTP-dependent reiterative transcription always occurs within the run of T residues in the initially transcribed region. The AUUUUn (where n = 1 to > 15) transcripts produced by this reaction are not extended productively to include downstream codBA sequences. In contrast, most transcripts initiated at position G7 do not engage in reiterative transcription and can be elongated normally. Characterization of a codBA promoter mutation that prevents reiterative transcription showed that this reaction is required for virtually all pyrimidine mediated regulation of operon expression and that UTP levels control the selection of the G7 and A8 transcriptional start sites. These results suggest a model for regulation in which high intracellular levels of UTP favor transcriptional initiation at position A8 and thus the accompanying reiterative transcription, which together preclude initiation at position G7. Low levels of UTP inhibit initiation at position A8 and the associated reiterative transcription, thereby allowing high levels of initiation at position G7 and operon expression. Our results also indicate critical sequence requirements for reiterative transcription, which are important for understanding the mechanism of this reaction as well as for identifying other promoters at which this reaction may occur. Of particular interest is the indication that an RNA:DNA hybrid forms during transcriptional initiation and the strength of this hybrid controls the extent of reiterative transcription. PMID- 7500335 TI - Determination of DNA persistence length by cryo-electron microscopy. Separation of the static and dynamic contributions to the apparent persistence length of DNA. AB - Axial deflection of DNA molecules in solution results from thermal motion and intrinsic curvature related to the DNA sequence. In order to measure directly the contribution of thermal motion we constructed intrinsically straight DNA molecules and measured their persistence length by cryo-electron microscopy. The persistence length of such intrinsically straight DNA molecules suspended in thin layers of cryo-vitrified solutions is about 80 nm. In order to test our experimental approach, we measured the apparent persistence length of DNA molecules with natural "random" sequences. The result of about 45 nm is consistent with the generally accepted value of the apparent persistence length of natural DNA sequences. By comparing the apparent persistence length to intrinsically straight DNA with that of natural DNA, it is possible to determine both the dynamic and the static contributions to the apparent persistence length. PMID- 7500334 TI - Adjacent upstream superhelical writhe influences an Escherichia coli promoter as measured by in vivo strength and in vitro open complex formation. AB - This work investigates the effect on transcription of superhelical writhe located in the region immediately upstream of the -35 consensus sequence of Escherichia coli promoters. A set of double-stranded oligonucleotides, each with an unique DNA configuration, were designed, synthesized and substituted into an area of naturally occurring right-handed superhelical curvature immediately upstream of the beta-lactamase promoter in plasmid pUC19. All the mutants showed reduced promoter activities in E. coli cells. However, rightward superhelical writhe clearly facilitated transcription when compared with the effect produced by a straight DNA segment. Leftward writhe greatly repressed transcription. A plane curve showed an intermediate effect. This phenomenon was due not only to the difference in the ability of the segment to drive an open complex formation, but also to the difference in the binding affinity of RNA polymerase to the promoter. The positive effect of rightward writhe was also observed in vivo for the promoter of the tetracycline resistance gene of pBR322. The sense and extent of superhelical writhe of a DNA curvature seem to determine its influence on promoter activity. PMID- 7500336 TI - Mutations in the gene encoding the 34 kDa subunit of yeast replication protein A cause defective S phase progression. AB - The in vivo function of the 34 kDa subunit of yeast replication protein A (RPA), encoded by the RFA2 gene, has been studied by analyzing the effect of Rpa34 depletion and by producing and characterizing rfa2 temperature-sensitive mutants. We show that unbalanced stoichiometry of the RPA subunits does not affect cell growth and cell cycle progression until the level of Rpa34 becomes rate-limiting, at which point cells arrest with a late S/G2 DNA content. Rpa34 is involved in DNA replication in vivo, since rfa2 ts mutants are defective in S phase progression and ARS plasmid stability, and rfa2 pol1 double mutants are non viable. Moreover, when shifted to the restrictive temperature, about 50% of the rfa2 mutant cells rapidly die while traversing the S phase and the surviving cells arrest in late S/G2 at the RAD9 checkpoint. Finally, rfa2 mutant cells have a mutator and hyper-recombination phenotype and are more sensitive to hydroxyurea and methyl-methane-sulfonate than wild-type cells. PMID- 7500338 TI - The structure of a novel DNA duplex formed by human centromere d(TGGAA) repeats with possible implications for chromosome attachment during mitosis. AB - The solution structure of the DNA duplex [GTGGAATGGAAC]2 containing a tandem repeat of the human centromere (TGGAA)n unit has been determined by two dimensional nuclear magnetic resonance (2D-NMR), distance geometry (DG) and molecular dynamics/energy minimization (MD/EM) methods. This remarkably stable "self-complementary" antiparallel duplex contains a tandem repeated motif in which unpaired guanine residues from opposite strands intercalate and costack between sheared G.A pairs. Twelve independent refined structures were determined from the NMR data and found to converge to a single family of closely related structures with pair-wise r.m.s.d. values of 0.55 +/- 0.25 Angstrum. All sugar residues are in the normal C2'-endo conformation except for the unpaired guanosines, which are in the unusual C3'-endo conformation. The guanosine residues of the bracketing G.A pairs have high-antiglycosidic torsion angles and zeta backbone torsion angles close to the trans domain. The structure exhibits many unusual interstrand interactions, including base-sugar stacking, base phosphate hydrogen bonding and cross-strand base stacking. The [GGA]2 unit contains a stack of four contiguous guanine residues, all of which have their hydrogen-bonding surface (N2H-N1H-O6-N7) exposed to solvent and available for interaction with other bases or ligands. This unexpected property may explain the unique morphology and function of the human centromere in mitosis. PMID- 7500337 TI - Dissection of the core and auxiliary sequences in the vegetative replication origin of promiscuous plasmid RK2. AB - The vegetative replication origin (oriV) of promiscuous IncP plasmid RK2 can function in many Gram-negative bacterial species when supplied with the plasmid encoded replication protein TrfA and host-encoded replication proteins including DnaA. Nine TrfA binding sites (iterons) are known, and also two DnaA binding sites, box 1, between TrfA iterons 4 and 5, and box 2, downstream of repeat 9. The deletion analysis presented here shows that the core oriV requires DnaA box 1 for function in Escherichia coli and Pseudomonas putida. This DnA box is not essential in Pseudomonas aeruginosa, although its deletion does reduce plasmid copy number in this species. A putative IHF binding site is located upstream of DnaA box 1, but IHF deficiency in E. coli seems not to alter replication efficiency or copy number control. Cloned oriV can interfere with maintenance of an independent RK2 replicon. Analysis of replication inhibition functions associated with oriV showed that a short putative orf between TrfA iterons 1 and 2 is not necessary for replication inhibition, the presence of repeats 5 to 9 in target and inhibitor plasmid are not sufficient for efficient inhibition and inhibition does not correlate directly with the number of direct repeats present. Rather, the results showed that the isolated repeats 1 and 2 to 4, potentiate replication inhibition disproportionately to their effect on the number of TrfA binding sites. The results are consistent with the idea that repeats 1 to 4, arranged as a single copy and as an irregular group of three, potentiate the ability of the oriV region to form complexes which inhibit replication. We suggest that TrfA bound at these iterons may be more susceptible to forming pairs between oriV sequences on different plasmids. PMID- 7500339 TI - Solution structure of a DNA quadruplex containing the fragile X syndrome triplet repeat. AB - Both X-ray and NMR structural studies have defined the polymorphic nature of G quadruplexes generated through mutual stacking of G.G.G.G tetrads by guanine rich telomeric sequences. Recently, the fragile X syndrome d(C-G-G)n triplet nucleotide repeat has been shown to form a stable quadruplex of undefined structure in monovalent cation solution. We have undertaken a structural characterization of the d(G-C-G-G-T3-G-C-G-G) undecanucleotide to elucidate the structural alignments associated with quadruplex formation by this oligomer which contains sequence elements associated with the fragile X syndrome triplet repeat. d(G-C-G-G-T3-G-C-G-G) in Na+ cation solution forms a quadruplex through dimerization of two symmetry related hairpins with the lateral connecting T3 loops positioned at opposite ends of the quadruplex. This novel NMR-molecular dynamics based solution structure contains internal G.C.G.C tetrads sandwiched between terminal G.G.G.G tetrads. Watson-Crick G.C base-pairs within individual hairpins dimerize through their major groove edges using bifurcated hydrogen bonds to form internal G(anti).C(anti).G(anti).C(anti) tetrads. Adjacent strands are anti-parallel to each other around the symmetric G-quadruplex which contains two distinct narrow and two symmetric wide grooves. By contrast, the terminal G tetrads adopt G(syn).G(anti).G(syn).G(anti) alignments. The structure of the d(G C-G-G-T3-G-C-G-G) quadruplex with its multi-layer arrangement of G.G.G.G and G.C.G.C tetrads greatly expands on our current knowledge of quadruplex folding topologies. Our results establish the pairing alignments that can be potentially utilized by the fragile X syndrome triplet repeat to form quadruplex structures through dimerization of hairpin stems. The formation of novel G.C.G.C tetrads through dimerization of Watson-Crick G.C base-pairs is directly relevant to the potential pairing alignments of helical stems in genetic recombination. PMID- 7500340 TI - Crystal structure of a bZIP/DNA complex at 2.2 A: determinants of DNA specific recognition. AB - The X-ray structure of the GCN4-bZIP protein bound to DNA containing the ATF/CREB recognition sequence has been refined at 2.2 A. The water-mediated interactions between the basic domain and DNA are revealed, and combined with a more accurate description of the direct contacts, further clarify how binding specificity is achieved. Water molecules extend the interactions of both invariant basic domain residues, asparagine 235 and arginine 243, beyond their direct base contacts. The slight bending of the basic domain alpha-helix around the DNA facilitates the linking of arginine 241, 243 and 245 to main-chain carbonyl oxygen atoms via water molecules, apparently stabilizing interactions with the DNA. PMID- 7500341 TI - Three-dimensional dimer structure of the lambda-Cro repressor in solution as determined by heteronuclear multidimensional NMR. AB - The 1H, 15N and 13C magnetic resonances of the lambda-Cro repressor have been assigned almost completely, mainly through the use of heteronuclear multidimensional NMR methods. Inter-subunit NOEs were distinguished by means of heteronuclear spectral editing technique (13C double half filter technique). Based on the distance and dihedral angle constraints derived from the NMR data, the three-dimensional solution structure of the lambda-Cro repressor in the dimeric form has been calculated by the simulated annealing method. The input for the structure calculations consisted of 1H-1H distance constraints, of which 1536 were intra-subunit and 40 were inter-subunit, and dihedral angle, phi, constraints, which numbered 92. The average root-mean-square deviation (RMSD) for all backbone heavy- atoms of the 20 calculated structures for residues 3 to 59 of the total of 66 amino acid residues in both subunits was 1.57 Angstrum, while the average RMSD for each subunit in the same residue range was 0.66 Angstrum. The subunit is composed of three alpha-helices, residues 7 to 13, 16 to 23 and 27 to 36, and a three-stranded anti-parallel beta-sheet composed of residues 3 to 6, 40 to 44 and 50 to 55. The solution structure of the subunit is essentially the same as that in the crystalline form, but the structure of the dimer form in solution differs from that of the dimer unit in the crystalline form. It is suggested that the solution dimer structure is distorted to fit the recognition helices in the major grooves of DNA on complex formation. PMID- 7500342 TI - Solution structure of the GTPase activating domain of alpha s. AB - We have used heteronuclear three-dimensional NMR spectroscopy to determine the solution structure of a 141 residue protein containing the GTPase activating domain from the alpha chain of the heterotrimeric G protein Gs. The domain contains six alpha-helices and is stable and structured in solution despite having been excised from the intact Gs protein. The N-terminal ten and C-terminal 11 residues of the protein are unstructured in solution while the core is well determined by the 2483 distance and torsion restraints derived from the NMR spectra. The final ensemble of 14 structures, generated with a hybrid distance geometry/simulated annealing protocol, have an average to-the-mean backbone root mean-square deviation of 0.39 A for the core residues 89 to 201. The majority of the structure is remarkably similar to that observed for the cognate domains in crystal structures of the homologous proteins alpha t and alpha i1. However, the orientations of the second helix and the subsequent interhelical loops differ markedly among the three proteins. This structural divergence, along with functional studies of chimeric proteins, suggests that this region of the domain interacts with either the downstream effector adenylyl cyclase or with some other intermediary protein. PMID- 7500343 TI - Solution structure of the HU protein from Bacillus stearothermophilus. AB - The histone-like protein HU from Bacillus stearothermophilus is a dimer with a molecular mass of 19.5 kDa that is capable of bending DNA. An X-ray structure has been determined, but no structure could be established for a large part of the supposed DNA-binding beta-arms. Using distance and dihedral constraints derived from triple-resonance NMR data of a 13C/15N doubly-labelled HU protein 49 distance geometry structures were calculated, which were refined by means of restrained Molecular Dynamics. From this set a total of 25 refined structures were selected having low constraint energy and few constraint violations. The ensemble of 25 structures display a root-mea-square co-ordinate deviation of 0.36 A with respect to the average structure, calculated over the backbone heavy atoms of residues 2 to 54 and 75 to 90 (and residues 2' to 54' and 75' to 90' of the second monomer). The structure of the core is very similar to that observed in the X-ray structure, with a pairwise r.m.s.d. of 1.06 A. The structure of the beta-hairpin arm contains a double flip-over at the prolines in the two strands of the beta-arm. Strong 15N-NH heteronuclear nuclear Overhauser effects indicate that the beta-arm and especially the tip is flexible. This explains the disorder observed in the solution and X-ray structures of the beta-arm, in respect of the core of the protein. Overlayed onto itself the beta-arm is better defined, with an r.m.s.d. of 1.0 A calculated over the backbone heavy atoms of residues 54 to 59 and 69 to 74. The tip of the arm adopts a well-defined 4:6 beta-hairpin conformation similar to the iron co-ordinating beta-arms of rubredoxin. PMID- 7500344 TI - Refined solution structure and dynamics of the DNA-binding domain of the heat shock factor from Kluyveromyces lactis. AB - The solution structure of the 92 residue (11 kDa) winged helix-turn-helix DNA binding domain from the kluyveromyces lactis heat shock factor was refined using a total of 932 NOE, 35 phi, 25 chi 1, 5 chi 2 and 44 hydrogen bond restraints. The overall root-mean-square deviation for structured regions was 0.75(+/- 0.15) A. The three-helix bundle and four-stranded beta-sheet are well defined with rmsd of 0.53(+/- 0.10) A and 0.60(+/- 0.17) A, respectively. Helix H2 is underwound and bent near Pro45. The angle between helix H2 and the proposed recognition helix H3 is 96(+/- 6) degrees. Detailed comparisons are made with the X-ray structure of this protein as well as other structural studies on HSF. Overall, the results are consistent with the earlier studies. Differences are related to protein-protein interactions in the crystal and dynamics in solution. Backbone dynamics was investigated via 15N relaxation. The average R1, R2 and NOE values for residues in segments of secondary structure were 1.9(+/- 0.9) s-1, 7.8(+/- 0.9) s-1 and 0.81(+/- 0.05), respectively. The correlation time based on these data was 5.6(+/- 0.4) ns. Motional order parameters were calculated by fitting the relaxation data to one of three models. Low-order parameters were found for residues that comprise the turn between helices H2 and H3 (residues Lys49 to Phe53), and most strikingly, the 16 residue wing (residues Val68 to Arg83). These data are consistent with the lack of long-range NOEs identified in these regions. The data provide a basis for comparison with results of the protein-DNA complex. The relationship between structure and function is discussed. PMID- 7500345 TI - Kinetic and structural characterization of mutations of glycine 216 in alpha lytic protease: a new target for engineering substrate specificity. AB - Gly216 in the active site of the broadly specific MA190 mutant of alpha-lytic protease has been found to be remarkably tolerant of amino acid substitutions. Side-chains as large as Trp can be accommodated within the substrate-binding pocket without abolishing catalysis, and have major effects upon the substrate specificity of the enzyme. Kinetic characterization of eleven enzymatically active mutants against a panel of eight substrates clearly revealed the functional consequences of the substitutions at position 216. To understand better the structural basis for their altered specificity, the GA216 + MA190 and GL216 + MA190 mutants have been crystallized both with and without a representative series of peptide boronic acid transition-state analog inhibitors. An empirical description and non-parametric statistical analysis of structural variation among these enzyme: inhibitor complexes is presented. The roles of active site plasticity and dynamics in alpha-lytic protease function and substrate preference are also addressed. The results strongly suggest that substrate specificity determination in alpha-lytic protease is a distributed property of the active site and substrate molecule. PMID- 7500346 TI - High helical propensity of the peptide fragments derived from beta-lactoglobulin, a predominantly beta-sheet protein. AB - It is generally assumed that peptide fragments derived from globular proteins are either unfolded, or adopt native-like secondary structures, in particular alpha helix, which are similar to those occurring in the early stages of protein folding. Since the structured conformations of short peptides are unstable, 2,2,2 trifluoroethanol (TFE) is often used to stabilize them. To examine the folding process of beta-proteins, we synthesized three fragments of beta-lactoglobulin corresponding to two beta-strands and one helix region, and examined their conformation by circular dichroism and nuclear magnetic resonance. These regions were chosen because, according to secondary structure prediction, all three should have high helix propensities. In aqueous solution, the three peptides had only a little ordered structure, but when TFE was added, they exhibited marked helical propensities, as also observed for the whole molecule of beta lactoglobulin. These results indicate that the intrinsic helical propensity of a peptide fragment elucidated by the addition of TFE is not necessarily related to the secondary structure in the native state. The results further suggest a case of non-hierarchical protein folding model, in which non-native structures may be involved in the early stage of folding. PMID- 7500347 TI - Vibrational Raman optical activity of alpha-lactalbumin: comparison with lysozyme, and evidence for native tertiary folds in molten globule states. AB - Proteins in aqueous solution are now accessible to Raman optical activity (ROA) measurements, which provide an incisive new probe of secondary and tertiary structure illustrated here by a study of bovine alpha-lactalbumin. The room temperature ROA spectrum of native bovine alpha-lactalbumin is similar to that of native hen egg-white lysozyme except for features attributable to differences in the loop regions: in particular, a positive ROA band at approximately 1338 cm-1 assigned to conformationally homogeneous loop structure, possibly with local order corresponding to 3(10)-helix, has more than double the intensity in alpha lactalbumin compared with lysozyme. This is consistent with the two proteins having similar secondary structure but different local details in the tertiary fold. ROA measurements on alpha-lactalbumin at pH 2.0 over a range of temperatures have provided a new perspective on the molten globule state. Thus at 35 degrees C ROA reveals the presence of some secondary structure but an almost complete loss of the tertiary loop structure; whereas at 2 degrees C the ROA spectrum is almost identical with that of the native protein, which is strong evidence that virtually all of the secondary structure and the tertiary backbone fold persist, albeit within a looser framework associated with increased solvent exposure and change of environment of many of the side-chains as evidenced by an increase in noise and bandwidth of some of the ROA signals together with aromatic fluorescence and near-UV circular dichroism signals characteristic of the molten globule state. Our sample of acid alpha-lactalbumin at 2 degrees C therefore appears to be an archetypal example of Ptitsyn's "native-like" molten globule, having a fixed native-like tertiary fold but with loss of tight packing of the side-chains; whereas at 35 degrees C it is a "disordered" molten globule. At 20 degrees C the acid molten globule appears to retain highly native-like secondary structure but with most of the tertiary fold already lost. A calcium-free sample of alpha-lactalbumin at neutral pH displayed a broad cooperative transition between native and molten globule states at approximately 15 degrees C, with the latter state showing similar but somewhat degraded tertiary loop ROA signatures to the native protein. In both the acid and apo molten globule states the ROA signatures of the secondary structure and the tertiary loops showed a gradual change with temperature.